PMID- 15108701 TI - Medical discipline--alternative medicine. PMID- 15108702 TI - Systemic Th1- and Th2-gene signals in atopy and asthma. AB - BACKGROUND: Atopic disorders have been associated with a Th-2 cytokine predominance. This study investigated Th1- and Th2-related gene expression in asthmatics, atopics and healthy individuals. METHODS: We compared Th1- and Th2 related in vivo-signals using gene expression arrays in 18 atopic asthmatics, 8 atopic non-asthmatic and 14 healthy control subjects. Purified mRNA from peripheral blood mononuclear cells was reverse-transcribed and hybridised to cDNA membranes. Group differences were assessed after standardisation with Mann Whitney U-test. RESULTS: Atopic individuals had upregulated lymphotoxin-alpha and downregulated IFNGR1. On the other hand, they had particularly high IL-4, IL-5 and IL4R levels, together with significantly upregulated IL10. Asthmatic individuals had normal Th1-gene expression, but an upregulation og Th-2 genes. Atopic individuals had high, asthmatic individuals excessively high IL12RB1 levels. No Th-2 gene was downregulated in both atopic phenotypes. The expression of IL6R correlated with the daily dose of inhaled corticosteroids. CONCLUSIONS: Atopic individuals had a down regulation of key TH1- and Th2-genes, resulting in a balanced upregulation of Th-specific genes. In contrast, asthmatic subjects had normal Th1-gene expression but a constant upregulation of Th2-specific genes, leading to Th2-predominance. PMID- 15108703 TI - Soft X-ray-induced decomposition of amino acids: an XPS, mass spectrometry, and NEXAFS study. AB - Decomposition of five amino acids, alanine, serine, cysteine, aspartic acid, and asparagine, under irradiation with soft X rays (magnesium Kalpha X-ray source) in ultra-high vacuum was studied by means of X-ray photoelectron spectrometry (XPS) and mass spectrometry. A comparative analysis of changes in XPS line shapes, stoichiometry and residual gas composition indicates that the molecules decompose by several pathways. Dehydration, decarboxylation, decarbonylation, deamination and desulfurization of pristine molecules accompanied by desorption of H2, H2O, CO2, NH3 and H2S are observed with rates depending on the specific amino acid. NEXAFS spectra of cysteine at the carbon, oxygen and nitrogen K-shell and sulfur L2,3 edges complement the XPS and mass spectrometry data and show that the exposure of the sample to an intense soft X-ray synchrotron beam results in the formation of C-C and C-N double and triple bonds. Qualitatively, the amino acids studied can be arranged in the following ascending order of radiation stability: serine 18) was used as a lifecourse factor. RESULTS: Mental health, proportion of friends who smoke and age of smoking initiation had strong associations with smoking status. However, they accounted for a small part of the association of being a lone mother and a smoker. After controlling for these factors, the odds of being a smoker among lone mothers were still twice those of mothers with partners (OR 2.1, 95% CI 1.7 2.7). CONCLUSIONS AND IMPLICATIONS: Improving the socio-economic status (SES), mental health and the social environment of lone mothers could help reduce their high smoking prevalence. However, much of the effect of being a lone mother remains even after controlling for these factors. More research is needed to discover why prevalence is so high among this demographic group. PMID- 15108746 TI - Evaluation of a new diabetes screening method at the Derbarl Yerrigan Health Service. AB - BACKGROUND AND OBJECTIVES: Diabetes is a major health problem for the Aboriginal population of Australia. Early detection is a key strategy to reduce the burden of diabetes. The aim of this study was to assess the acceptability, sensitivity and specificity, effectiveness and cost of a new method of screening for diabetes at Derbarl Yerrigan, the Aboriginal health service in Perth. METHODS: Between January and May 1999, all clients over the age of 30 years and not known to have diabetes were approached for HbA1c testing using the DCA 2000 analyser. Those whose HbA1c results indicated the need for follow-up were asked to return for confirmatory testing using the gold standard oral glucose tolerance test (OGTT). A questionnaire was administered to participants who did not return. A file audit was conducted over 15 non-consecutive days to determine screening, follow-up and the number of new cases diagnosed using the previous ad hoc approach to screening with a glucometer. RESULT: 238 clients were approached and all agreed to participate and undertook the test. Of these, 37 were referred for follow-up diagnostic testing. Of these, only 14 had an OGTT. Among these 14, five were found to have diabetes and three were found to have impaired glucose tolerance. Of the remaining 23 participants, we were able to contact only six and administer the questionnaire to four. Poor follow-up meant that the sensitivity, specificity and cost-effectiveness of the test could not be assessed. CONCLUSIONS AND IMPLICATIONS: The reasons for poor follow-up need to be investigated if Aboriginal health services are to be more successful at screening for diabetes. PMID- 15108747 TI - Evaluation of immunisation coverage for aboriginal and Torres Strait Islander children using the Australian Childhood Immunisation Register. AB - OBJECTIVE: To estimate immunisation coverage for routinely administered vaccines among children using receipt of a particular Hib vaccine (PRP-OMP) as a proxy for Indigenous status. METHODS: Until May 2000, PRP-OMP was provided only for Indigenous children in all jurisdictions except the Northern Territory. In three one-year ACIR-derived birth cohorts, any child recorded on the ACIR as receiving one or more doses of PRP-OMP as the only Hib vaccine was presumed to be Aboriginal and Torres Strait Islander. Using this proxy, estimated numbers of Indigenous children were compared with Australian Bureau of Statistics estimates, and immunisation status for recommended vaccines was estimated at 12 and 24 months by jurisdiction and remoteness compared with children who received other Hib vaccines (presumed non-Indigenous). RESULTS: The numbers of Aboriginal and Torres Strait Islander children estimated using this 'proxy method' are approximately 42% of those estimated by the ABS. Immunisation coverage (among proxy Indigenous children) at 12 months (72-76%) and 24 months (64-73%) was considerably lower than others (90-94% and 81-88%, respectively). These children had significantly lower coverage when living in accessible areas than remote areas. CONCLUSIONS AND IMPLICATIONS: These data provide the first national measure of immunisation status and are likely to be a valid measure among those identified. Aboriginal and Torres Strait Islander immunisation coverage is 17% lower with the biggest gaps in urban areas, indicating the need for better quality data informing appropriate interventions. PMID- 15108748 TI - Demographic and socio-economic factors associated with dental health among older people in NSW. AB - OBJECTIVE: To investigate the association between oral health status and social, economic and demographic factors in community-dwelling older people in New South Wales (NSW). METHODS: Binary and multinomial logistic regression analyses were used to examine the associations between measures of oral health status (edentulous/dentate, and the frequency of toothache or mouth or denture problems in the previous 12 months) and demographic and socio-economic factors using data from the NSW Older People's Health Survey 1999. RESULTS: After adjusting for other factors, being edentulous was associated with being older, having no private dental insurance, being female, leaving school at less than 15 years of age, not being financially comfortable, not being a homeowner, living in a rural area, and being unable to travel alone. Among both dentate and edentulous people, increasing age and being able to travel independently were associated with decreased reporting of toothache, mouth or denture problems; while not being financially comfortable was associated with increased reporting of toothache or mouth or denture problems. The frequency of mouth or denture problems was not found to be independently associated with having private dental insurance nor with holding a health concession card. CONCLUSIONS: Among older people in NSW, oral health is associated with a range of demographic and socio-economic factors. The results suggest that better oral health among older people is associated with a capacity to pay out-of-pocket dental expenses rather than with private dental insurance or having access to public-funded dental care. PMID- 15108749 TI - Patient co-payments and use of prescription medicines. AB - OBJECTIVE: To investigate how prescription co-payments influence the medicine use of Australian patients. METHODS: Two surveys and an in-depth interview study were conducted in the Newcastle/Hunter region of New South Wales (NSW). A community based survey explored how often prescription cost posed a barrier to prescription use. A general practice patient survey investigated the impact of prescription cost on the timing of medical consultations and prescription collection. Quantitative data were summarised using descriptive statistics; associations between household characteristics and outcomes were explored using odds ratios and chi square analysis. In-depth interviews were conducted to explore the role of prescription cost in medicine use. The interview data were qualitatively analysed for relevant themes using 'grounded theory'. RESULTS: 420 of 950 households (44%) participated in the community survey: 110 (26%) reported delaying visiting a GP, 85 (20%) not buying all of their prescription medicines and 77 (18%) not refilling a prescription because of cost. Sixty-two (15%) households reported significant difficulties with prescription costs. Households with children had twice the odds of reporting significant difficulties than those without (OR= 2.0, 95% CI 1.2-3.5). Of the 442 (43%) GP patients who participated, 25 (6%) patients reported prescription cost as the reason for delaying their visit. Of the 291 patients who received a prescription, 26 (9%) patients reported cost as the reason for not collecting some or all of their prescriptions. IMPLICATIONS: Given the wide variation in patients' capacity to manage increased out-of-pocket costs, co-payments may add to patients' burden and place a potential barrier to safe and timely prescription use. PMID- 15108750 TI - Getting it right--the Australian Childhood Immunisation Register and immunisation rates in south-eastern Sydney. AB - OBJECTIVES: To assess the completeness of ACIR data for south-eastern Sydney children at 12 to < 15 months of age and to develop strategies to improve rates. METHODS: We surveyed children aged 12 to < 15 months listed as overdue on the ACIR011A report, December 2001, by contacting the last immunisation provider or the parents. RESULTS: From the 470 children listed as overdue, 162 children were systematically selected to form the study group. Seventeen were lost to follow-up and results are available for 145 children. Eighty children were up to date (69, encounter forms incorrect or not sent; 11, vaccinated overseas) and 11 children had moved overseas. Fifty-four children were overdue (30, provider error due to incorrect Haemophilus influenzae type b (Hib) schedule or Hib vaccine omitted; 18 did not complete schedule; 6 conscientious objectors). CONCLUSIONS: Our study indicates that between 6-9% of children on the register were likely to be overdue, whereas the ACIR011A report identified 19%. Failure of immunisation providers to correctly complete, or send encounter forms to the ACIR was the main reason for this discrepancy. Migration and failure to record overseas vaccination were also factors. IMPLICATIONS: Public health units should develop good working relationships with immunisation providers to assist and encourage immunisation and the completion and submission of encounter forms. A quarterly review, using the third dose assumption, of all children aged 12 to < 15 months identified on the ACIR011A report as overdue for immunisation may be an effective way for public health units to increase apparent rates for their area. PMID- 15108751 TI - Predictors of incomplete immunisation in Victorian children. AB - OBJECTIVE: Using probabilistic record linkage, this study aimed to determine the predictors of incomplete immunisation in Victorian children. METHODS: Records of all births in Victoria in 1998 were linked with records from the Australian Childhood Immunisation Register (ACIR). The outcomes of interest were the predictors of immunisation status at 12 and 24 months of age. RESULTS: A total of 91.1% of birth records were linked with records from the ACIR (n=55,129). Immunisation coverage was 92.8% at 12 months and 89.2% at 24 months. After multivariate analysis, the following maternal factors were significant predictors of incomplete immunisation in children aged 12 months: higher parity, 12-23 months interval between pregnancies, not being married, Aboriginal or Torres Strait Islander or born overseas, younger age, no private health insurance, home birth, metropolitan place of birth, and being in the highest or lowest socioeconomic quintiles. Low birthweight and singleton birth were also significant predictors. All of these factors were also predictive of incomplete immunisation in children aged 24 months, with the exception of low birth weight, which was only significant in the very low birth weight category. Congenital malformations or prematurity were not significant predictors. CONCLUSIONS AND IMPLICATIONS: Record linkage is an effective means of determining the relationship between immunisation status and socio-demographic and health-related factors. It enabled small groups within a large population to be studied, confirming findings from previous survey studies and indicating that several population groups were at increased risk of incomplete immunisation. The main limitation of the study was the under-reporting of immunisation status to the ACIR. PMID- 15108752 TI - To what extent is health and medical research funding associated with the burden of disease in Australia? AB - OBJECTIVE: This study aimed to analyse the association of the National Health and Medical Research Council fund allocations and several measures of burden of disease in Australia, and compare it to similar studies in the United States and Canada. METHODS: A cross-sectional study comparing disease-specific funding in two time periods (1998-2001 and 2002-03) with data from the Australian Burden of Disease study on four measures of burden of disease (incidence, mortality, years of life lost and disability-adjusted life years in 1996). This association was measured by correlation coefficients. With the use of these measures as predictor variables in a regression analysis, predicted funding was calculated and compared with actual funding. RESULTS: The highest correlation coefficients (r = 0.68 0.75) were exhibited by the DALYs and years of life lost to disability and the relation was significant at p < 0.0001 (1998 to 2003). Based on DALYs, the top five under-funded categories (1998-2001) were intentional injuries, cardiovascular diseases, mental disorders, unintentional injuries and chronic respiratory diseases. The top five over-funded categories were infectious and parasitic diseases, nervous system and sense organ disorders, malignant neoplasms, endocrine and metabolic disorders and genitourinary diseases. CONCLUSIONS: This study revealed a significant relation between NHMRC research funding and burden of disease measures and highlighted that comparison of actual and predicted funding based on different measures of disease can alter conclusions as to whether a disease is over- or under-funded. PMID- 15108753 TI - Some issues to consider when assessing concordance of death certificates with registry reports. PMID- 15108754 TI - Norovirus excretion in a healthcare worker without major symptoms of gastroenteritis: infection control implications. PMID- 15108755 TI - Solaria use by minors in Australia: is there a cause for concern? PMID- 15108756 TI - Consulting with Pacific people: a palangi steps out. PMID- 15108757 TI - Evaluating end-of-life opioid use in managed care plans. PMID- 15108758 TI - Opioid use and health care charges at the end of life in patients with metastatic cancer. AB - Opioid use and health care charges during the final year of life in patients with metastatic cancer who received controlled-release oxycodone (CRO), transdermal fentanyl (TF), or controlled-release morphine sulfate (CRM) were examined in the following study. A total of 704 patients were identified who met study entry criteria; CRO, TF, or CRM was the first-received long-acting opioid (LAO) during the last year of life for 36% (N = 250), 29% (N = 203), and 36% (N = 251) of study subjects, respectively. On average, patients initiated LAO therapy three to four months before death. One-half of patients received less than 60 days of LAO therapy. Mean total health care charges in the final year of life were dollars 84,572 for patients receiving CRO, dollars 90,935 for patients receiving TF, and dollars 76,446 for patients receiving CRM. Patients receiving TF started LAO therapy closer to their date of death, and had significantly higher hospitalization charges than did patients receiving CRO and CRM. PMID- 15108759 TI - The role of the health care ombudsman. AB - Beginning in the 1990s, with increased attention focused on the alleged abuses and arbitrary decision making by MCOs, several states began enacting legislation to establish health care ombudsmen. A variety of states, including the District of Columbia, are now considering health care ombudsman legislation. This article focuses on the varying role, functions, and duties of the health care ombudsman in each of these states and their benefit to the health care environment. PMID- 15108760 TI - Emerging therapies for angina: new modalities may offer additional clinical options. AB - The social and economic burdens of chronic angina are enormous, and the morbidity and mortality associated with its underlying cause, coronary artery disease, are high. For many patients with chronic angina, current drug therapies are contraindicated or ineffective, and surgical procedures too risky or costly. Most newer approaches have not yet been sufficiently evaluated and can be expensive, time-consuming, or both. A new class of antianginal drugs, the metabolic modulators, will soon be available. In controlled clinical trials, these agents have shown promising results in reducing the frequency of angina episodes and lengthening the duration of exercise sessions. They are also well tolerated. PMID- 15108761 TI - The cost of chronic obstructive pulmonary disease and its effects on managed care. AB - Chronic obstructive pulmonary disease (COPD) designates respiratory disorders characterized by airway obstruction that is not fully reversible. An estimated 10 million adult Americans have COPD, and the prevalence is rising. Direct and indirect costs of managing COPD exceed dollars 32 billion annually, and this health care burden has provoked vigorous efforts by major public health organizations to evaluate and improve quality of care for COPD. The authors review the substantial effects of COPD on managed care and discuss evidence-based strategies for its effective management. PMID- 15108762 TI - The effect of tiered-copay formularies on prescription drug use and spending. PMID- 15108763 TI - Bibliography. Current world literature. Protein and amino acid metabolism. PMID- 15108764 TI - End-stage renal failure and cardiac mortality after heart transplantation. AB - BACKGROUND: Coronary artery disease (CAD) is the leading cause of mortality after the first year of heart transplantation. End-stage renal failure (ESRF) is more frequent because of long-term survival. Impact of ESRF on cardiac mortality in heart transplant patients is unappreciated. The hypothesis of accelerated CAD in uremic patients has been suggested. METHODS: In Pitie La Salpetriere hospital, 1211 heart transplants have been performed between 1982 and 2001. Thirty-three patients have reached ESRF. A case-control study was performed to identify risk factors responsible for ESRF and to appreciate the impact of ESRF on cardiac mortality. RESULTS: In cases at 6 months, serum creatinine tended to be higher (159 +/- 31 micromol/L vs. 141 +/- 44 micromol/L, p = 0.06) and cyclosporine (CSA) dosage (mg/kg) was significantly lower (5.4 +/- 1.8 mg/kg vs. 7.7 +/- 2.7 mg/ kg, p = 0.002). Mean triglyceride level after transplantation until dialysis was significantly lower in cases (2.18 +/- 0.82 mmol/L vs. 1.48 +/- 0.62 mmol/L, p = 0.002). In cases and controls, cardiac mortality was responsible for 67% (10 of 15) and 38% (three of eight) of all deaths, respectively. High triglyceride level (> or = 2 mmol/L) was associated with cardiac mortality [p < 0.03, hazard ratio (HR) = 3.89]. Kaplan Meier cardiac free survival rates were significantly lower in cases than in controls (p < 0.03). CONCLUSION: These data suggest that CSA nephrotoxicity could result from individually determined susceptibility and that hypertriglyceridemia may have a negative impact on renal function and cardiac mortality. The risk of cardiac mortality is increased in heart transplant patients with ESRF. The hypothesis of accelerated atherosclerosis in ESRF patients after heart transplantation leading to higher cardiac mortality incidence needs further study. PMID- 15108765 TI - Impact of in vivo complement activation and cryoglobulins on graft outcome of HCV infected renal allograft recipients. AB - BACKGROUND: Chronic hepatitis C virus (HCV) infection is closely associated with mixed cryoglobulinemia. Cryoglobulins can activate complement leading to vascular damage. We examined whether cryoglobulinemia and complement turnover is associated with HCV infection in renal transplant recipients and whether this has an adverse effect on graft outcome. METHODS: Sera and fresh plasma from 31 HCV RNA-positive patients after renal transplantation (group I) were studied for cryoglobulins, complement hemolytic activity (CH50), and complement split product C3d. In total, 80 HCV-negative renal transplant recipients (group II) and 72 untreated patients with chronic hepatitis C (group III) without renal transplantation served as controls. RESULTS: Cryoglobulins were detected in 45, 28, and 26% of the patients in group I, II, and III, respectively. A high cryocrit ( > 5%) was present only in patients of group III (p < 0.01%). Mean CH50 values were lower and C3d levels higher in HCV-positive patients (group I and III) compared with HCV-negative patients (p < 0.0001). Cryoglobulins were not associated with extrahepatic manifestations or graft dysfunction, except in five patients of group III demonstrating cryoglobulinemic vasculitis. HCV-positive renal transplant recipients with signs of complement activation showed a significantly greater increase of serum creatinine (0.88 +/- 1.14 mg/dL) when compared with baseline than patients without complement activation (0.34 +/- 0.37 mg/dL; p = 0.035). There was also a tendency toward a higher extent of proteinuria in patients with complement activation (1.38 +/- 2.17 g/d vs. 0.50 +/ 0.77 g/d; p = 0.25, NS). CONCLUSIONS: Cryoglobulins are common in renal allograft recipients, but do not affect graft function. However, complement activation appears to be involved in chronic allograft dysfunction in HCV infected recipients. PMID- 15108766 TI - Activation of nuclear factor-kappa B and macrophage invasion in cyclosporin A-and tacrolimus-treated renal transplants. AB - This retrospective study was designed to compare the efficacy of cyclosporin A (CyA) and tacrolimus (FK506) on chronic rejection (CR) associated with nuclear factor-kappa B (NF-kappaB) activation and macrophage invasion. Non-episodic day 50 protocol renal biopsy was performed in 63 consecutive patients with renal transplants from living donors, treated with either CyA or FK506. Southwestern histochemistry for NF-kappaB, immunostaining for CD68, and Banff classification were performed, and these findings were compared with outcome over 34 +/- 13 months. Compared with specimens from FK506-treated patients (n = 20), specimens from CyA-treated patients (n = 43) showed a significant increase in tubulointerstitial CD68-positive cells (1.5 +/- 0.9 vs. 0.9 +/- 0.8, p < 0.01), although no significant differences were observed in NF-kappaB activation. Specimens with Banff acute rejection (AR) grade > or = 1A (n = 20) showed increased macrophages (p < 0.01) compared with specimens with AR < 1A (n = 43). Specimens from patients with clinical AR prior to day 50 biopsy (n = 23) also showed increased macrophage invasion (p < 0.01) compared with specimens from patients without prior clinical AR (n = 40). The cumulative well-functioning (serum creatinine < 1.5 mg/dL) graft survival rate was significantly lower in patients with increased tubulointerstitial CD68-positive cells (n = 63, p < 0.05). Our findings suggest that tacrolimus is more effective than CyA against CR with respect to macrophage invasion and AR. PMID- 15108767 TI - Delayed occurrence of cardiac allograft vasculopathy in Chinese heart transplant recipients compared with their western counterparts. AB - Cardiac allograft vasculopathy (CAV) is a leading limiting factor to long-term survival after cardiac transplantation. We investigated the prevalence of CAV and its associated factors in Chinese heart transplant recipients. From July 1987 to July 2000, we performed 140 consecutive heart transplantations at the National Taiwan University Hospital. Of the 140 patients, 98 who were > or = 17-yr old at the time of transplantation, had survived for more than 1 yr after transplantation, and who had normal findings at the 1-month coronary angiogram study, were included in this study. Group I consisted of 25 patients who eventually developed CAV in the follow-up, and group II consisted of 73 patients who were free from CAV in the follow-up. CAV was defined by coronary angiogram study.The donor and recipient characteristics were not statistically different between the two groups except the older donor age (p = 0.02), higher first-year mean rejection score (p = 0.03) and more prevalent cytomegalovirus infection rate (p = 0.03) in group I. Multivariate Cox regression analysis revealed that only higher first-year mean rejection score (p = 0.01), and older donor age (p = 0.04) were important risk factors for developing CAV. The 1-5 yr of actuarial freedom from the presence of CAV were 97, 93, 86, 80 and 69% in our study patients. In summary, these data show that CAV occurred later in Chinese heart transplant recipients in comparison with their western counterparts, but the risk factors for developing CAV were not different. PMID- 15108768 TI - An open-label randomized trial of the safety and efficacy of sirolimus vs. azathioprine in living related renal allograft recipients receiving cyclosporine and prednisone combination. AB - BACKGROUND: The ability of sirolimus (SRL), in combination with reduced exposure of cyclosporine, was investigated to prevent acute rejection and associated side effects. METHODS: Between June 1999 and February 2000, 70 recipients of primary one-haplotype living-related donor renal allografts were randomized to receive SRL (2 mg/d) or azathioprine (AZA) (2 mg/kg/d) combined with cyclosporine and prednisone. The primary end-point was a composite of first occurrence of biopsy confirmed acute rejection, graft loss, or death during the first 3 months after transplantation. RESULTS: From week 4 to month 12, SRL patients received lower cyclosporine (week 4: 364 mg/d vs. 455 mg/d, p = 0.004; month 12: 195 mg/d vs. 255 mg/d, p = 0.038) doses and showed lower cyclosporine concentrations (week 4: 247 ng/mL vs. 309 ng/mL, p = 0.04; month 12: 143 ng/mL vs. 188 ng/mL, p = 0.045). Compared with AZA, SRL patients showed reduced 3-month primary end point (0% vs. 17.1%, p = 0.025), and reduced incidence of biopsy-confirmed acute rejection at 3 months (0% vs. 14.3%, p = 0.01) but not at 12 months (11.4% vs. 14.3%, NS). Mean creatinine at 12 months were not different (1.8 +/- 0.6 vs. 1.6 +/- 0.6, p = 0.23). Hyperlipidemia was the only adverse event more frequent among SRL patients (49% vs. 17%, p = 0.01). There were no differences in infections and no malignancies in both groups. CONCLUSIONS: The combination of 2 mg fixed doses of SRL, reduced cyclosporine exposure and prednisone was associated with a low incidence of acute rejection and did not result in significantly impaired graft function compared with patients receiving AZA, standard doses of cyclosporine and prednisone. PMID- 15108769 TI - Increasing medication access to transplant recipients. AB - Transplant recipients require numerous medications to maintain graft survival and health. Post-transplant medication therapy costs greater than 12000 dollars annually, and the cost of therapy is expected to increase. Although medication costs continue to rise, a substantial portion of Americans lack adequate health insurance or do not have any insurance coverage. To facilitate health, it is imperative that health care providers are familiar with programs that are available to increase prescribed medication access to transplant recipients. The purpose of this manuscript is to provide an overview of common programs available to increase transplant recipients' access to medications. In addition to discussing the consequences of medication non-compliance, this manuscript reviews Medicare, Medigap, Medicaid, Qualified Medicare Beneficiaries, and pharmaceutical manufacturers' medication assistance programs and its use to increase medication access to solid-organ transplant recipients. PMID- 15108770 TI - The impact of pancreas transplantation on patient employment opportunities. AB - BACKGROUND: A questionnaire study was designed to determine whether the quality of life benefit provided by receiving a kidney-pancreas (KP) transplant increased the incentive to return to work, compared with patients receiving a kidney alone (KA) transplant. METHODS: A confidential questionnaire was sent to 58 patients with insulin-dependent diabetes mellitus who had functioning grafts at least 6 months post-transplant. Thirty-eight patients had received a KP transplant and 20 patients, a KA transplant. RESULTS: Eighty-three percent of patients returned the questionnaire (87% of KP and 75% of KA patients [p: non-significant (ns)]. The pre-transplant demographic variables of age, gender, educational status, and need for dialysis, were equivalent between the KP and KA groups, respectively (p: ns). At the time of transplantation 39% of KP patients and 33% of KA patients were employed at least 20 h/wk (p: ns). However, post-transplant significantly more KP patients (73%) were working compared with KA patients (27%, p: 0.04). Additionally, pre-transplant employment was independently associated with post transplant work status (p: 0.01). CONCLUSION: These data suggest that the improvement in quality of life associated with pancreas transplantation provide an added incentive for diabetics with renal insufficiency to seek employment. PMID- 15108771 TI - Observations regarding the use of sirolimus and tacrolimus in high-risk cadaveric renal transplantation. AB - BACKGROUND: Balancing the risk of acute rejection (AR) with drug-induced toxicities complicates the selection of the optimal immunosuppressive regimen, especially in the high-risk renal transplant recipient. This study was designed to determine the optimal dosage combinations of tacrolimus and sirolimus in a high-risk cadaveric renal transplant population. METHODS: Primary cadaveric renal transplant recipients were randomly assigned to receive either standard tacrolimus (trough levels of 10-15 ng/mL) plus reduced sirolimus (trough levels of 5-10 ng,mL) (Group I) or to receive reduced tacrolimus (trough levels of 5-10 ng,mL) plus standard sirolimus (trough levels of 10-15 ng/mL) (Group II). All patients received Thymoglobulin induction and steroids. RESULTS: Thirty-nine (16 in Group I and 23 in Group II) high-risk renal transplant recipients (100% cadaveric donors, 79% African-American recipients, and 59% delayed graft function) are the subjects of this report. At 6 months, the patient survival rate was 94 and 100% and the graft survival rate was 94 and 83% in Groups I and II, respectively. The incidence of biopsy-proven AR was 6 and 5% in Groups I and II, respectively. Eight patients (50%) in Group I required discontinuation of tacrolimus, seven because of biopsy-proven tacrolimus nephrotoxicity and one secondarily to interstitial pneumonitis. Wound complications were the most frequent adverse event reported in both groups. CONCLUSIONS: The combination of tacrolimus and sirolimus was associated with a low risk of AR in this cohort of high-risk renal transplant recipients. However, 50% of patients who received standard tacrolimus and reduced sirolimus combination had to be discontinued from the regimen because of biopsy-proven nephrotoxicity. These preliminary results provide evidence that sirolimus should not be added to tacrolimus without dosage adjustments. We have discontinued recruitment of patients to the standard tacrolimus and reduced sirolimus combination and we have tightened our criteria for selection of marginal donor kidneys with our high-risk renal transplant recipients. PMID- 15108772 TI - Long-term outcome of liver transplantation for autoimmune hepatitis. AB - BACKGROUND: Liver transplantation is the final therapeutic option for about 10% of patients with autoimmune hepatitis (AIH) who do not respond to medical therapy. The aim of this study was to evaluate the long-term outcome in serologically defined subgroups of AIH after transplantation. METHODS: Pre- and post-transplantation data of 28 patients with AIH transplanted between 1987 and 1999 were retrospectively analyzed and compared with 24 patients, who underwent liver transplantation because of Wilson's disease and glycogen storage disease type 1. RESULTS: Serological analyses identified patients with AIH type 1 (n = 13), type 2 (n = 5), and type 3 (n = 10). The 5-yr patient survival rate after liver transplantation was 78.2%, which was not significantly different from the control group. Six AIH patients and four control patients required re transplantation because of initial non-function, chronic rejection or AIH recurrence. Patients transplanted for AIH (88%) had more episodes of acute rejection when compared with patients transplanted for genetic liver diseases (50%). Clinical and histological features of chronic rejection were present in four patients, which did not differ significantly from the controls. Recurrence of AIH was diagnosed in nine patients (32%) based upon the presence of autoantibodies, increased gamma-globulins, steroid dependency, and histological evidence of chronic hepatitis. These combined features were not found in any of the controls. CONCLUSIONS: Our data do not suggest that AIH subtypes influence prognosis after liver transplantation. Despite a high frequency of acute cellular rejection episodes and disease recurrence, transplantation for AIH has a 5-yr survival rate, which does not differ from that observed in patients transplanted for genetic liver diseases. PMID- 15108773 TI - Comprehensive examination of gene expression associated with long-term stable graft acceptance by renal transplant recipients. AB - Expression levels of mRNA in peripheral blood mononuclear cells from five renal transplant recipients and five non-transplanted controls were analyzed with GeneChips (GeneChip Instrument system, Affymetrix, Santa Clara, CA, USA). All recipients had retained a well-functioning kidney graft for more than 15 yr on low-dose maintenance immunosuppression. Among a total of 12630 transcripts examined, significant differential expression was observed for 599 genes, whereby 470 genes were up-regulated and 129 down-regulated in the transplant recipients compared with controls. Of these, 192 up-regulated and 46 down-regulated genes showing a change greater than twofold were divided into eight functional categories as follows (numbers of genes, up/down): immune system (12/14), cell proliferation (17/3), oncology (15/3), transporter/receptor/binding protein (16/5), transcription factors (8/2), enzymes (17/4), expressed sequence tags (91/9), and others (16/6). Predictably, expression of immune-associated genes was decreased in the recipients. Significant reduction of expression levels of CD3, ICAM-1, and B7.2, which are critical molecules for interactions between antigen presenting cells and T cells, were observed. In T cell signal transduction, the Ras pathway was likely to be suppressed by activation of hVH-5. The present data help to elucidate the immunological status in long-term kidney graft recipients and may provide insights for future regimens to establish donor-specific hyporesponsiveness. PMID- 15108774 TI - Eversion thromboendovenectomy in organized portal vein thrombosis during liver transplantation. AB - Portal thrombosis is no longer considered a contraindication for transplantation because of the technical experience acquired in the field of liver transplantation and the development of various surgical techniques. All the same, the results obtained in portal thrombosis patients are at times suboptimal, and the surgical technique used (thromboendovenectomy or veno-venous bypass) is also controversial. Between May 1988 and December 2001, 455 liver transplants were performed, of which 32 (7%) presented portal vein thrombosis. Of these, eight belonged to the first 227 transplants (group I), and 24 to the other 228 (group II). Of the 32 cases with portal thrombosis, 20 (62%) were type Ib, seven (22%) type II/III and five (16%) type IV. Twenty-two were males (69%), with a mean age of 50 yr (range: 30-70 yr); the thrombosis in all cases developed over a cirrhotic liver: 15 cases of an ethanolic origin, 11 because of hepatitis C virus, two cases of autoimmune aetiology, one case of primary biliary cirrhosis, one case because of hepatitis B virus and two cases of a cryptogenic origin. Five cases had a history of surgical treatment for portal hypertension. The surgical method in all cases consisted of an eversion thromboendovenectomy (ETEV) under direct visual guidance, with occlusion of the portal flow using a Fogarty balloon. Once re-canalization was achieved, we performed local heparinization and end-to-end portal anastomosis. In no case was systemic post-operative heparinization performed. In the 32 cases in which thrombectomy was attempted it was achieved in 31 of them (96%), failing only in a case of type IV thrombosis, which was resolved by portal arterialization. Of the 31 successful cases, only one with type IV thrombosis re-thrombosed. The 5-yr survival rate of the patients in the series was 69%, with 10 patients dying, of whom only two from causes related to the thrombosis and the thrombosis treatment, both with type IV thrombosis. The ideal treatment for portal thrombosis during liver transplantation is controversial and depends on its extension and the experience of the surgeon. In our experience, ETEV resolves most thromboses (types I, II and III), but management of type IV, which occasionally can be treated with this technique, may require more complex procedures such as bypass, portal arterialization or cavoportal haemitransposition. PMID- 15108775 TI - Tube thoracostomy during allogeneic stem cell transplantation does not carry an increased risk for infections or bleeding. AB - BACKGROUND: Candidates for stem cell transplantation may occasionally suffer from massive pleural effusions related to their disease and require tube thoracostomy. The additional risk of this procedure during allogeneic transplantation procedure is not known. METHODS: Four high-risk patients transplanted in our institution during a 2-yr period had chest drainage by tube thoracostomy. The characteristics of the fluid, the clinical course, and the outcome were assessed. RESULTS: A total of nine chest drains were inserted (range 1-5). No bleeding complications related to the procedure were noted. None of the patients developed any clinical signs of local infection at the tube insertion site or within the pleural fluid. All cultures taken from the drained fluid or from the insertion wound were negative. CONCLUSIONS: Tube thoracostomy in itself does not seem to pose additional risks in the transplant procedure, despite all patients in this series being considered to be at high-risk for complications. PMID- 15108776 TI - Morbid obesity does not preclude successful renal transplantation. AB - Many renal transplantation centers arbitrarily deny transplantation to patients with morbid obesity usually defined as body mass index > 35. We present a series of 173 primary renal transplant patients in a new transplant program that accepted all recipients with 3 yrs or greater life expectancy and no active malignancy or infection. When the patient outcomes are divided into groups by body mass index, it can be seen as expected that patients with body mass index > 30 have an increased prevalence of wound infections (p < 0.05). However, aside from this complication there are no statistically significant outcome differences between the three groups realizing the possibility of type II statistical error because of small numbers. Graft survival, patient survival and other surgical complications are the same in all groups regardless of body mass index. At the end of the 3-yr interval with a minimum transplant follow-up of 3 months, 169 of 173 patients were alive and 163 of 173 transplants were functioning. Based on our experience, morbid obesity should not be used to exclude patients arbitrarily from transplantation anymore than advanced age or diabetes should. PMID- 15108777 TI - Post-liver transplant acute renal failure: factors predicting development of end stage renal disease. AB - BACKGROUND: Acute renal failure (ARF) occurs in 5-50% of patients undergoing orthotopic liver transplantation (OLT). The aim of this study was to determine factors that might predict the development of end stage renal disease (ESRD) in patients who had ARF after OLT. METHODS: We studied all OLT recipients between 9/1/1988 through 12/31/2000. RESULTS: A total of 1602 patients underwent OLT during the study period. About 350 patients (22%) developed ARF requiring dialysis post-operatively. One hundred and twenty-three (39.8%) died within a year after OLT. Median follow up was 5.8 yr (range 1-12 yr). Forty-three patients (23%) developed ESRD over median of 3.79 yr (range 1-8 yr). Multivariate logistic regression analysis revealed creatinine levels > 1.7 mg/dL at 1 yr (p < 0.001), cyclosporine as immunosuppression (p = 0.026), and the presence of diabetes pre OLT (p < 0.001) to be associated with the development of ESRD. The development of ESRD did not decrease patient survival (p = 0.111). ESRD patients who received subsequent kidney transplantation had significantly improved survival rates (p = 0.005). CONCLUSIONS: Serum creatinine levels at 1 yr, cyclosporine as immunosuppression, and the presence of diabetes pre-OLT are independent predictive factors for the development of ESRD. ESRD patients who received kidney transplantation had higher 10-yr survival rates when compared with patients maintained on dialysis. PMID- 15108778 TI - Cytomegalovirus ischemic colitis of a diabetic renal transplant recipient. AB - We report a diabetic renal transplant recipient with cytomegalovirus (CMV) disease who presented with tarry stool diarrhea because of multiple colonic ulcerations. Histopathology revealed diffuse colonic ulcers following a process of ischemic vasculitis. The colonic ulcers disappeared dramatically after 2 wk of intravenous ganciclovir therapy. Hyper-immunosuppression was initially suspected but acute rejection (AR) developed after immunosuppressive reduction during the ganciclovir therapy. The AR was successfully reversed and the dosage of cyclosporine was returned to the same level prior to the onset of CMV disease. Our experience suggests that ganciclovir is quite effective for healing colonic ulcers caused by CMV and acute allograft rejection may occur during therapy. PMID- 15108779 TI - A new indication for pancreas transplantation: high grade pancreatic dysplasia. AB - A 42-yr-old male presented with a family history of pancreatic carcinoma inherited an autosomal dominant pattern. The development of endocrine and exocrine pancreatic insufficiency served as early markers for neoplastic transformation. Screening endoscopic ultrasound and ERCP showed abnormalities suggestive of pancreatic dysplasia. Total pancreatectomy was performed and pathology confirmed carcinoma in situ, also known as high-grade pancreatic ductal dysplasia or Pan IN-3. The patient's post-operative course was complicated by life threatening, brittle diabetes. Pancreas transplantation was successfully performed. One year following transplantation, the patient has excellent pancreas graft function. He remains insulin free and has no signs of malignancy. Total pancreatectomy followed by pancreas transplantation is a viable therapeutic option for patients in the dysplastic but still pre-malignant phase of familial pancreatic adenocarcinoma who develop hypoglycemic unawareness following total pancreatectomy. PMID- 15108780 TI - Massive gastrointestinal hemorrhage due to rupture of a donor pancreatic artery pseudoaneurysm in a pancreas transplant patient. AB - Enteric drainage of secretions by anastomosing the donor duodenum to the recipient's small bowel has become common in pancreatic transplantation. While it eliminates many problems, endoscopic access to the transplanted duodenum and pancreas is made difficult. After a pancreas kidney transplant, the patient presented with massive hematochezia. Upper and lower endoscopy revealed large amounts of red blood in the colon but no specific bleeding site. Mesenteric angiography was normal but pelvic angiography showed rapid extravasation of contrast from a pseudoaneurysm of the pancreatic transplant artery. This was successfully embolized with coils. To the best of our knowledge, this is the first case of massive gastrointestinal hemorrhage because of rupture of a pseudoaneurysm of the donor pancreatic artery in a pancreas transplant patient. We report this case and review our institution's experience with all forms of gastrointestinal bleeding in pancreas transplant patients. PMID- 15108781 TI - Predictors of failure to cure atrial fibrillation with the mini-maze operation. AB - BACKGROUND: Maze-III is a complex surgical procedure designed to treat chronic atrial fibrillation. A reduction in the number of right and left atrial incisions could decrease the operative time. The aim of this study was to assess the results of a mini-maze operation and to define predictors of its failure. METHODS: Between 1995 and 2000, 72 patients (mean age 64 +/- 9 years) undergoing cardiac surgery had a concomitant mini-maze operation for symptomatic chronic atrial fibrillation. Three and 12 months post-operatively, heart rhythm and left atrial transport functions were assessed by electrophysiology, echocardiography, and magnetic resonance imaging. Multivariate analysis was performed to identify predictors of failure of the mini-maze operation. RESULTS: Operative mortality was 1.4% (1/72). Death during follow-up occurred in 5.6% of patients (4/71), in one due to chronic heart failure. After 1 year, 80% of patients (48/60) were either in sinus rhythm (n = 43; 72%) or had a pacemaker (n = 5; 8%) implanted due to sick sinus syndrome. Intermittent and chronic atrial fibrillation was found in 20% of patients (12/60). Preoperative duration of atrial fibrillation (p = 0.05), preoperative left atrial diameter (p = 0.001), preoperative right atrial diameter (p = 0.02), a reduced left ventricular ejection fraction (p = 0.03), an increased left ventricular end-diastolic diameter (p = 0.04), and the presence of mitral valve stenosis (p = 0.001) were found to be univariate predictors of failure of the mini-maze operation 1 year postoperatively. Multivariate analysis defined preoperative diagnosis of mitral valve stenosis (p = 0.005; OR 117.5), longer duration of preoperative atrial fibrillation (p = 0.01; OR 1.33), and increased preoperative left ventricular end-systolic diameter (p = 0.02; OR 1.2) as incremental independent risk factors for failure of the mini-maze operation to cure chronic atrial fibrillation. CONCLUSION: The mini-maze operation is a safe procedure with similar results to that of Cox's Maze-III operation. The less invasive mini-maze operation could be applicable even to patients with severely reduced left ventricular function, in whom complex cardiac surgery has to be performed concomitantly as well as in those presenting severe comorbidities. PMID- 15108782 TI - Open heart surgery in patients 85 years and older. AB - BACKGROUND: There has been an increase in the number of elderly patients considered for cardiac surgery. Several reports have documented acceptable morbidity and mortality in patients 80 years and older. The results from surgical patients 85 years and older were analyzed. METHODS: The records of 89 consecutive patients 85 years and older having cardiac operations between June 1993 and May 1999 were retrospectively reviewed. For purposes of statistical analysis follow up was considered as a minimum of one office visit to the surgeon, cardiologist, or internist at least 1 month postoperatively. RESULTS: Eighty-seven patients underwent coronary artery grafting and two patients had mitral valve replacement. Follow-up was 100% complete. The operative mortality rate was 12.3%; probability of in-hospital death was 8.2%; risk-adjusted mortality rate was 3.2%. The complication rate was 31.5%. The actuarial 1-, 3-, and 5-year survivals were as follows: 75%, 67%, and 40%. Multivariate predictors of 30-day mortality were preoperative EF, less than 30% (p = 0.029) and postoperative renal failure (p = 0.0039). CONCLUSIONS: Cardiac surgery can be performed in patients 85 years and older with good results. There is an associated prolonged hospital stay for elderly patients. Consistent successful outcomes can be expected in this patient population with selective criteria identifying risk factors. PMID- 15108783 TI - The clinical outcome and quality of life following minimally invasive direct coronary artery bypass surgery. AB - BACKGROUND: Minimally invasive direct coronary artery bypass (MIDCAB) through a limited anterior small thoracotomy has been shown to be a promising technique of surgical treatment for single or double vessel disease. Little is known about the Health-Related Quality Of Life (HRQOL) in this group of patients. METHODS: The records of 75 consecutive patients who underwent MIDCAB procedure at Harefield Hospital between April 2000 and January 2002 were reviewed retrospectively. HRQOL assessment was planned in a cross-sectional design. Patients were contacted by telephone to conduct a semi-structured interview and were sent two questionnaires: the Short Form health survey (SF-36) and the Hospital Anxiety and Depression Scale (HADS). RESULTS: There was no in-hospital death. Patients stayed in the Intensive Therapy Unit (ITU) for 11.56 +/- 4.55 hours and stayed in hospital for 3 +/- 2.34 days. None of the study patients had perioperative myocardial infarction (MI) or neurological complications including permanent and transient strokes. We were able to contact all the 75 patients by telephone and they also completed the SF-36 and HADS. The SF-36 scores were compared to an age matched group of normal British people. The MIDCAB group had an excellent general health perception compared to the normal group (p < 0.001), but similar scores otherwise. The HADS scores showed that only 1 patient (1.3%) had mild depression, 5 patients (6.7%) had mild anxiety, and 2 patients (2.6%) had moderate anxiety. CONCLUSION: MIDCAB is a safe surgical treatment and provides excellent clinical and HRQOL outcomes. PMID- 15108784 TI - Timing of replacement therapy for acute renal failure after cardiac surgery. AB - OBJECTIVE: Acute renal failure (ARF) following cardiac surgery remains a significant cause of mortality. The aim of this study is to compare early and intensive use of continuous veno-venous hemodiafiltration (CVVHDF) with conservative usage of CVVHDF in patients with ARF after cardiac surgery. MATERIALS AND METHODS: Due to ARF, CVVHDF was required in two groups of a total of 61 adult patients (1.79% of all patients). Group 1 included 27 patients while Group 2 included 34 patients. CVVHDF was performed on Group 1 when creatinine level exceeded 5 mg/dL, or potassium level exceeded 5.5 mEq/L irrespective of the urine output. CVVHDF was performed on Group 2 when urine output was less than 100 mL within consecutive 8 hours, with no response to 50 mg furosemide with the supplementary criterion that urine sodium concentration should be >40 mEq/L before the administration of furosemide. RESULTS: The mean elapsed time between the surgery and the initiation of CVVHDF was 2.56 +/- 1.67 days in Group 1 and 0.88 +/- 0.33 days in Group 2 (p = 0.0001). The mean intensive care unit (ICU) stay for Group 1 was 12 +/- 3.44 days and 7.85 +/- 1.26 days for Group 2 (p = 0.0001). ICU mortality rate was 48.1% for Group 1 and 17.6% for Group 2 (p = 0.014). The overall hospital mortality rate was 55.5% for Group 1 and 23.5% for Group 2 (p = 0.016). CONCLUSION: Recognition of ARF and early beginning of the CVVHDF are extremely important. The sooner the ARF after surgery is recognized and CVVHDF is performed, the higher the likelihood of the reduction of the hospital mortality. PMID- 15108785 TI - Regional cardiac sympathetic innervation early and late after transmyocardial laser revascularization. AB - BACKGROUND: Prior experimental and clinical studies have drawn disparate conclusions regarding the effects of transmyocardial laser revascularization (TMR) on regional cardiac innervation in the treated regions. Regional afferent denervation has been proposed as a potential mechanism of action of the procedure, although this as yet remains unproven. The purpose of the present study was to evaluate regional myocardial sympathetic innervation both early (3 days) and late (6 months) after TMR. METHODS: Mini-swine in the early group were randomized to be sacrificed 3 days after holmium:YAG TMR (n = 5) or sham thoractomy (n = 3). In the late group, mini-swine with hibernating myocardium in the left circumflex (LCx) region were randomized to sham redo-thoracotomy (n = 5), TMR of the LCx distribution with a carbon dioxide (n = 5), holmium:YAG (n = 5), or excimer (n = 5) laser. Six months postoperatively the animals were sacrificed. Additional animals in both the early (n = 2) and late (n = 2) groups served as age- and weight-matched normal controls. Immunohistochemistry and Western blot analysis for tyrosine hydroxylase (TYR-OH), a neural-specific enzyme found in sympathetic efferent nerves and a commonly used anatomic marker of regional innervation, were performed on lased and nonlased LCx and septal regions. RESULTS: Immunohistochemical staining for TYR-OH was markedly diminished in the lased myocardial regions 3 days after TMR. This staining was significantly reduced compared to untreated septal regions, sham-operated, and normal LCx myocardium. Quantitative immunoblotting confirmed a significant reduction in TYR OH (p < 0.05) protein concentration in the lased regions 3 days after TMR. On the contrary, TYR-OH staining was present in LCx myocardium surrounding the laser channels of all animals in all groups 6 months postoperatively. Staining was not different from controls. Similarly, there was no difference in LCx TYR-OH protein concentration between the normal, sham, or 6 months postoperative lased groups (p > 0.2 by one-way ANOVA). CONCLUSIONS: TMR-treated myocardium demonstrates anatomic evidence of regional sympathetic denervation 3 days postoperatively, although myocardium lased with each of the three lasers currently in clinical use is reinnervated by 6 months as evidenced by immunoblotting and immunohistochemistry for TYR-OH. These results suggest that mechanisms other than denervation may account for the long-term reductions in angina seen after TMR. PMID- 15108786 TI - Is maintenance of cerebral hypothermia the principal mechanism by which retrograde cerebral perfusion provides better brain protection than hypothermic circulatory arrest? A study in a porcine model. AB - OBJECTIVE: Retrograde cerebral perfusion (RCP) provides better brain protection than hypothermic circulatory arrest (HCA) alone. The mechanism by which RCP improves brain protection during circulatory arrest remains unknown. The purpose of the study in pigs was to determine if RCP improves brain protection mainly as a result of its ability to maintain cerebral hypothermia. METHODS: Fifteen pigs were subjected to 120 minutes of HCA alone (HCA group, n = 5), HCA + RCP at perfusion pressures of 23 to 29 mmHg (RCP-low group, n = 5), or at perfusion pressures of 34-40 mmHg (RCP-high group, n = 5) at 15 degrees C, followed by 60 minutes of normothermic cardiopulmonary bypass (CPB). After brain temperature reached 15 degrees C, HCA was initiated with or without RCP. Temperatures in the brain, esophagus, and perfusate/blood were monitored continuously. Brain tissue blood flow was measured continuously using a laser flowmeter. Brain oxygen extraction was calculated from the oxygen contents in arterial and venous blood samples. RESULTS: During cooling and rewarming, the change in temperature was slower in the brain than in the esophagus. A similar degree of spontaneous rewarming (from 15 degrees C to 17/18 degrees C) occurred in the brain during HCA and RCP. This indicates that RCP does not provide better maintenance of cerebral hypothermia during circulatory arrest than HCA alone. The esophageal temperature rose more slowly during RCP than during HCA alone, indicating that RCP maintains better hypothermia in the body. During RCP, the brain extracted oxygen continuously from the blood, indicating that RCP may provide nutrient flow to the brain. CONCLUSION: In an acute pig model, maintenance of cerebral hypothermia does not appear to be the principal mechanism by which RCP provides better brain protection than HCA alone. Retrograde cerebral perfusion provides nutrient flow/oxygen to brain tissue, leading to better brain protection than HCA alone. PMID- 15108787 TI - Coronary artery bypass surgery in diabetic patients. AB - BACKGROUND: Cardiovascular disease is a major cause of morbidity and mortality in patients with diabetes. This study examines the impact of diabetes on mortality and morbidity following coronary artery bypass surgery. METHODS: We retrospectively analyzed 590 consecutive patients after coronary artery bypass grafting in 1998. Reoperations and combined procedures were excluded. A total of 137 diabetic (23.2%) and 453 nondiabetic patients were evaluated. Among the diabetics, 53 were treated with insulin and 84 were non-insulin-dependent. Diabetics suffered more frequently from hypertension, peripheral vascular disease, and more often had an increased body mass index (BMI). RESULTS: There was no significant difference in mortality and major complications among insulin dependent diabetics, non-insulin-dependent diabetics, and nondiabetic patients. Diabetics suffered more often from superficial sternal wound infection and had a higher incidence of superficial wound infections at the vein harvest site. CONCLUSION: The present study suggests that diabetes increases the risk of superficial wound infections after coronary artery bypass grafting. But diabetics do not necessarily have an increased risk of major complications and mortality. PMID- 15108788 TI - Abnormal origin of the right subclavian artery from the right pulmonary artery in a patient with D-transposition of the great vessels and left juxtaposition of the right atrial appendage: an unusual anatomical variant. AB - Isolation of the left subclavian artery (LSCA) or its anomalous origin from the pulmonary artery (PA) has been documented in several cases, especially in association with a right-sided aortic arch. Similar anomalies involving the right subclavian artery (RSCA) are less frequent. Anomalous origin of the RSCA from the PA in association with D-transposition of the great arteries (D-TGA) is exceedingly rare and only two cases have been reported so far. We present here, a case of aberrant origin of the RSCA from the right PA in a patient with D-TGA, in whom the diagnosis was rendered difficult due to the partial occlusion of the intervening ductus arteriosus (DA). We discuss the embryological basis of this anomaly and review its clinical and surgical implications. PMID- 15108789 TI - Transbrachial coil occlusion of the large branch of an internal mammary artery coronary graft. AB - Percutaneous transbrachial insertion of two complex coils into the intercostal branch of the left internal mammary artery resulted in the relief of severe angina in a 45-year-old man who had coronary artery bypass surgery 2 years before. The diagnosis of coronary artery steal was made clinically. This case illustrates the importance of recognizing coronary steal in patients who redevelop angina after coronary artery surgery with the use of an incompletely prepared left internal mammary artery as a conduit. Brachial or radial artery should be preferred to reach left internal mammary artery (LIMA) for cannulation easily. The preoperative angiographic imaging of LIMA is important to detect the side branches and their sizes. The patient was treated without the need for further surgery. PMID- 15108790 TI - Explantation of a mechanical assist device: assessment of myocardial recovery. AB - Reliable predictors of myocardial recovery post-mechanical assist device implantation are not well defined. We report the case of a 29-year-old woman with refractory congestive heart failure due to acute myocarditis of unknown etiology. The patient was supported with a HeartMate (Thoratec Corp., Woburn, MI) left ventricular assist device for 100 days showing improvement in clinical status. Hemodynamic monitoring, dobutamine stress echocardiography, and cardiopulmonary testing were performed to identify myocardial recovery. Improvements in myocardial reserve, hemodynamic parameters, and myocardial oxygen consumption were identified prior to explantation. The patient was successfully explanted. This case confirms that myocardial recovery is feasible with the use of a mechanical assist device. This article describes our current left ventricular assist device weaning protocol. PMID- 15108791 TI - Redo-OPCAB via left thoracotomy using symmetry aortic connector system: a report of two cases. AB - Left thoracotomy is an established approach for redo coronary artery bypass grafting (CABG). This approach has also been successfully used in off-pump coronary artery bypass (OPCAB). Traditionally, the grafts have been anastomosed proximally to the descending thoracic aorta or the left subclavian artery. Recently, proximal connectors have been introduced by various manufacturers for use on ascending aorta during primary CABG and OPCAB. One such device is the Symmetry aortic connector system (St. Jude Medical, Minneapolis, MN). These devices have obviated the need for partial occluding clamps for the construction of the proximal anastomoses and hence are extremely useful when the aorta is heavily calcified. We used this device successfully in two patients undergoing redo-OPCAB, where the proximal anastomosis was constructed on the descending aorta. In so doing, we also used the shortest possible length of vein graft since the descending aorta at that level was much closer than the left subclavian artery. This can be an additional factor in redo-operations where the availability of vein can be an issue. PMID- 15108792 TI - Dor operation for a young male with left ventricular aneurysm due to spontaneous left anterior descending coronary artery dissection. AB - Surgical cases for myocardial ischemia due to spontaneous coronary artery dissection have rarely been reported. We describe a young male who had a myocardial infarction with left ventricular aneurysm due to spontaneous left anterior descending coronary artery dissection. He was successfully treated with Dor's left ventriculoplasty without coronary artery revascularization. The Dor procedure was a simple and effective treatment. To our knowledge, this is the first report in which the Dor procedure was used to treat spontaneous coronary artery dissection with left ventricular aneurysm. PMID- 15108793 TI - Beating heart aortic valve replacement in a pregnant patient. AB - A 23-year-old woman with a history of previous bileaflet mechanical aortic valve replacement presented in acute heart failure at 27 weeks of gestation. Transthoracic echocardiography (TEE) demonstrated minimal leaflet excursion consistent with valve thrombosis. The patient underwent emergent surgery with normothermic cardiopulmonary bypass (CPB), and beating-heart technique for valve replacement. This novel strategy for aortic valve replacement is being presented as being particularly useful in pregnant patients whose fetuses are at risks of hypothermia, hemodilution, and hyperkalemia during CPB. PMID- 15108794 TI - Repair of left coronary artery aneurysm, recurrent ascending aortic aneurysm, and mitral valve prolapse 19 years after Bentall's procedure in a patient with Marfan syndrome. AB - A 45-year-old female with Marfan syndrome had a Bentall's procedure performed 19 years ago. She presented with a 4-year history of gradually worsening dyspnea and decreasing exercise tolerance. Investigations revealed severe mitral valve prolapse, a left main stem coronary artery (LMSCA) aneurysm, and a recurrent aneurysm of the ascending aorta. The mitral valve was replaced and the aortic aneurysmal sac and the LMSCA aneurysm were then repaired by a modified Bentall procedure. The patient made an uneventful recovery and was discharged home. PMID- 15108795 TI - The use of an aortic connector system in a patient with severe calcified ascending aorta. AB - Patients with severe calcified ascending aorta ("porcelain aorta") present a surgical challenge. Off-pump coronary artery bypass grafting (OPCAB), using new automated proximal anastomotic devices, provide a surgical alternative for patients who are not candidates for in situ arterial grafting. We present a 74 year-old male with double-vessel disease and a large calcified aneurysm of the left anterior wall. Left ventricular function was poor with an ejection fraction of 24%. The beating heart technique was used for the distal anastomosis and ventriculoplasty. The proximal anastomosis was constructed with an automated aortic connector system, thereby avoiding clamping of the severely diseased aorta. PMID- 15108796 TI - John H. Gibbon, Jr., the inventor of the first successful heart-lung machine. AB - May 6th, 2003 marked the 50th anniversary of the first successful use of the heart-lung machine. It was an event that would dramatically change the field of cardiac surgery and the approach to the treatment of cardiac disease. It was also the culmination of years of work of Dr. John H. Gibbon, Jr. We take a look at the life of this remarkable man. PMID- 15108797 TI - Spontaneous right ventricular disruption following treatment of sternal infection. AB - BACKGROUND: Spontaneous right ventricular disruption is a rare and frequently catastrophic event that occurs during the treatment of mediastinitis complicating median sternotomy wound. OBJECTIVE: The purpose of this study is to understand the pathogenesis of the spontaneous right ventricular disruption and to suggest strategies for the prevention and treatment of this rare but potentially fatal complication of cardiac surgery. METHODS: We report three cases as an introduction to the review of 39 cases found in the English-language literature. RESULTS: The majority of patients (71%) underwent coronary artery bypass grafting as the primary procedure prior to the development of a sternal infection. Staphylococcus Aureus and Staphylococcus Epidermidis were cultured most frequently from the sternal wound (31% and 24%, respectively). The mean interval between sternal debridement and the right ventricular disruption was 2.9 days. Most patients (24 of 42) required cardiopulmonary bypass for the repair of the right ventricular disruption. Biologic patches and adjuncts were used in 15 patients (36%). Eight patients (19%) died either preoperatively or on the operating-room table. CONCLUSIONS: Spontaneous right ventricular disruption is a potentially preventable complication. To prevent this complication we recommend: (1) avoidance of delay between diagnosis and operative treatment of mediastinitis; (2) complete lysis of adhesions between the posterior sternal edge and anterior surface of the right ventricle under general anesthesia with heart lung machine stand-by; (3) repair of the right ventricular tear using biologic patches with heart-lung machine stand-by; (4) early (if possible immediate) closure of the chest with a myocutaneous flap. PMID- 15108798 TI - The role of endogenous and exogenous DNA damage and mutagenesis. AB - The field of DNA damage responsiveness in general, and the consequences of endogenous and exogenous base damage in DNA, in particular, has made new and exciting contributions to our increasing understanding of the initiation and progression of neoplasia in humans. This article presents some of the highlights in this area of investigation, with a particular emphasis on DNA repair, the tolerance of DNA damage and its contribution to mutagenesis, and DNA damage checkpoint regulation. PMID- 15108799 TI - DNA damage tumor suppressor genes and genomic instability. AB - Disruption of the mechanisms that regulate cell-cycle checkpoints, DNA repair, and apoptosis results in genomic instability and the development of cancer in multicellular organisms. The protein kinases ATM and ATR, as well as their downstream substrates Chk1 and Chk2, are central players in checkpoint activation in response to DNA damage. Histone H2AX, ATRIP, as well as the BRCT-motif containing molecules 53BP1, MDC1, and BRCA1 function as molecular adapters or mediators in the recruitment of ATM or ATR and their targets to sites of DNA damage. The increased chromosomal instability and tumor susceptibility apparent in mutant mice deficient in both p53 and either histone H2AX or proteins that contribute to the nonhomologous end-joining mechanism of DNA repair indicate that DNA damage checkpoints play a pivotal role in tumor suppression. PMID- 15108800 TI - MCM proteins: DNA damage, mutagenesis and repair. AB - The MCM2-7 complex, which may act as a replicative helicase during DNA synthesis, plays a central role in S-phase genome stability. MCM proteins are required for processive DNA replication and are a target of S-phase checkpoints. Loss of MCM function causes DNA damage and genome instability. MCM expression is upregulated in proliferating cells, providing a diagnostic marker for both cancerous cells and cells with the potential to become malignant. The role of the MCM complex in genome integrity reflects its activity both at active replication forks and away from forks. PMID- 15108801 TI - Those dam-aged telomeres! AB - Telomere integrity plays a crucial role in the capacity for continuous cell proliferation. In some circumstances, shortened telomeres contribute to cell arrest or death, but in others, shortened telomeres may actually enhance the incidence and spectrum of tumors. Resolution of this apparent paradox requires a more detailed understanding of a non-functional telomere. Recent evidence reveals that critically shortened or uncapped telomeres share molecular hallmarks of damaged DNA. It is likely that the cellular response to this DNA damage, influenced by the nature of the damage itself, affects the outcome of loss of telomere function. PMID- 15108802 TI - Aurora kinases link chromosome segregation and cell division to cancer susceptibility. AB - Aurora kinases play critical roles in chromosome segregation and cell division. They are implicated in the centrosome cycle, spindle assembly, chromosome condensation, microtubule-kinetochore attachment, the spindle checkpoint and cytokinesis. Aurora kinases are regulated through phosphorylation, the binding of specific partners and ubiquitin-dependent proteolysis. Several Aurora substrates have been identified and their roles are being elucidated. The deregulation of Aurora kinases impairs spindle assembly, checkpoint function and cell division, causing missegregation of individual chromosomes or polyploidization accompanied by centrosome amplification. Aurora kinases are frequently overexpressed in cancers and the identification of Aurora A as a cancer-susceptibility gene provides a strong link between mitotic errors and carcinogenesis. PMID- 15108803 TI - Reversibility of oncogene-induced cancer. AB - Cancer can largely be conceived as a consequence of genomic catastrophes resulting in genetic events that usurp physiologic function of a normal cell. These genetic events mediate their pathologic effects by either activating oncogenes or inactivating tumor-suppressor genes. The targeted repair or inactivation of these damaged gene products may counteract the effects of these genetic events, reversing tumorigenesis and thereby serve as an effective therapy for cancer. However, because they are the result of many genetic events, the inactivation of no single mutant gene product may be sufficient to reverse cancer. Despite this caveat, compelling recent evidence suggests that there are circumstances when even the brief interruption of activation of a single oncogene can be sufficient to reverse tumorigenesis. Understanding how and when oncogene inactivation reverses cancer will be important in both defining the molecular pathogenesis of cancer as well as developing new molecularly based treatments. PMID- 15108804 TI - Therapeutic implications of cancer stem cells. AB - Most cancers comprise a heterogenous population of cells with marked differences in their proliferative potential as well as the ability to reconstitute the tumor upon transplantation. Cancer stem cells are a minor population of tumor cells that possess the stem cell property of self-renewal. In addition, dysregulation of stem cell self-renewal is a likely requirement for the development of cancer. This new model for cancer will have significant ramifications for the way we study and treat cancer. In addition, through targeting the cancer stem cell and its dysregulated self-renewal, our therapies for treating cancer are likely to improve. PMID- 15108805 TI - Multiple niches for Notch in cancer: context is everything. AB - Notch receptor signaling has very distinctive roles in cancers originating from different types of cells that reflect its complex functions in normal tissue development and homeostasis. For example, recent studies have shown that Notch signals are oncogenic in pre-T cells but suppress tumor development in keratinocytes. Notch signaling contributes to pre-malignant metaplastic changes that precede pancreatic carcinoma, and it is also likely to be involved in other forms of metaplasia. In addition, several viral oncoproteins and chromosomal translocations target one or more components of a Notch transcriptional activation complex. PMID- 15108806 TI - New roles for the RB tumor suppressor protein. AB - For a gene whose existence was first postulated in 1971, was cloned in 1986 and whose functions have been extensively characterized ever since, you might be inclined to think there was not much new to report regarding the retinoblastoma tumor suppressor gene (RB)--but you would be wrong to make such an assumption. RB is still piquing our interest with several activities defined over the past year that reveal new and exciting roles for this key tumor suppressor gene. These functions include regulation of senescence through specific gene silencing mechanisms, control of developmental processes in extra-embryonic tissues, maintaining tissue homeostasis and determining survival responses to chemotherapy. PMID- 15108807 TI - Ski and SnoN: negative regulators of TGF-beta signaling. AB - Ski and SnoN are unique proto-oncoproteins in that they can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. Recent studies using in vitro and in vivo approaches have begun to unravel the complex roles of Ski and SnoN in tumorigenesis and embryonic development. The identification of Ski and SnoN as important negative regulators of signal transduction by the transforming growth factor-beta superfamily of cytokines provides a valuable molecular basis for the complex functions of Ski and SnoN. PMID- 15108808 TI - Death in the third dimension: apoptosis regulation and tissue architecture. AB - Tissue development, homeostasis and tumor pathogenesis all depend upon a complex dialogue between multiple cell types operating within a dynamic three-dimensional (3D) tissue extracellular matrix microenvironment. A major issue is whether the spatial organization of a cell within this 3D tissue microenvironment could modulate cell responsiveness to regulate cell fate decisions such as survival, and if so how. Classic developmental model systems and transgenic animals are instructive but pose special challenges for investigators conducting signaling studies and biochemical assays in tissues. As an alternative, 3D culture model systems exist in which cell-adhesion dependent tissue architecture, heterotypic cell-cell interactions and tissue differentiation can be recapitulated with good fidelity. 3D cell culture models are slowly revealing how tissue architecture can dramatically influence how a cell responds to exogenous stimuli to modify its apoptotic behavior and hence should prove instrumental for identifying novel cell death pathways. PMID- 15108809 TI - HIF-1 and hypoxic response: the plot thickens. AB - Hypoxia, or lowered physiological oxygenation, is a component of many diseases. The response to decreased oxygen tensions is also an essential aspect of normal development and physiology. The hypoxic response is chiefly mediated by a single transcription factor--the hypoxia-inducible transcription factor HIF-1. The central role played by HIF-1 in hypoxia-induced transcription has made the focus of intense investigation by biologists from a wide range of disciplines. Recent developments have produced fundamental advances in our understanding of both the regulation and function of this response pathway in animals, and illustrated the potential for manipulation of the pathway for therapeutic purposes. PMID- 15108810 TI - Genes that drive invasion and migration in Drosophila. AB - Successful cell migration depends on the careful regulation of the timing of movement, the guidance of motile cells, and cytoskeletal and adhesive changes within the cells. This review focuses on genes that act cell-autonomously to promote these aspects of cell migration in Drosophila. We discuss recent advances in understanding the migration of the ovarian border cells, embryonic blood cells, primordial germ cells, somatic gonadal precursors, and tracheal cells. Comparison of genes that regulate these processes to those that promote tumorigenesis and metastasis in mammals demonstrates that studies in fruit flies are uncovering new genes highly relevant to cancer biology. PMID- 15108811 TI - Multiple connections link FAK to cell motility and invasion. AB - The ability of intracellular signaling networks to orchestrate a complex biological response such as cell motility requires that individual signaling proteins must act as integrators, responding to multiple extracellular inputs and regulating multiple signaling pathway outputs. In this review, we highlight recent findings that place focal adhesion kinase (FAK) in an important receptor proximal position in the regulation of growth factor and integrin-stimulated cell motility. Emphasis is placed on the molecular mechanisms of FAK activation, connections of FAK to focal contact formation as well as turnover, and the potential that FAK function in promoting cell invasion may be distinct from its role in cell motility. PMID- 15108812 TI - Proximodistal patterning of the limb: insights from evolutionary morphology. AB - There is an active debate about how skeletal elements are encoded along the proximodistal (PD) axis of the developing limb. Our aim here is to see whether consideration of the evolutionary morphology of the limb can contribute to our understanding of patterning mechanisms. Of special interest in this context are animals showing reiterated skeletal elements along the PD axis (e.g., dolphins and plesiosaurs with hyperphalangy). We build on previous hypotheses to propose a two-step model of PD patterning in which specification of broad domains in the early limb bud is distinct from subsequent processes that divides an initial anlage into a segmental pattern to yield individual skeletal elements. This model overcomes a major evolutionary problem with the progress zone model, which has not previously been noted: pleiotropy. Parallels with other developmental systems are briefly discussed. PMID- 15108813 TI - How different types of pattern formation mechanisms affect the evolution of form and development. AB - Here we investigate how development and evolution can affect each other by exploring what kind of phenotypic variation is produced by different types of developmental mechanisms. A limited number of developmental mechanisms are capable of pattern formation in development. Two main types have been identified. In morphodynamic mechanisms, induction events and morphogenetic processes, such as simple growth, act at the same time. In morphostatic mechanisms, induction events happen before morphogenetic mechanisms, and thus growth cannot influence the induction of a pattern. We present a study of the variational properties of these developmental mechanisms that can help to understand how and why a developmental mechanism may become involved in the evolution and development of a particular morphological structure. Using existing models of pattern formation in teeth, an extensive simulation analysis of the phenotypic variation produced by different types of developmental mechanisms is performed. The studied properties include the amount and diversity of the phenotypic variation produced, the complexity of the phenotypic variation produced, and the relationship between phenotype and genotype. These variational properties are so different between different types of mechanisms that the relative involvement of these types of mechanisms in evolutionary innovation and in different stages of development can be estimated. In addition, type of mechanism affects the tempo and mode of morphological evolution. These results suggest that the basic principles by which development is organized can influence the likelihood of morphological evolution. PMID- 15108814 TI - Inductive interactions and embryonic equivalence groups in a basal metazoan, the ctenophore Mnemiopsis leidyi. AB - Ctenophores undergo locomotion via the metachronal beating of eight longitudinally arrayed rows of comb plate cilia. These cilia are normally derived from two embryonic lineages, which include both daughters of the four e1 micromeres (e11 and e12) and a single daughter of the four m1 micromeres (the m12 micromeres). Although the e1 lineage is established autonomously, the m1 lineage requires an inductive interaction from the e1 lineage to contribute to comb plate formation. Successive removal of the e1 progeny at later stages of development indicates that this interaction takes place after the 32-cell stage and likely proceeds over a prolonged period of development. Normally, the e1, cell lies in closest proximity to the m12 cell that generates comb plate cilia; however, either of the e1 daughters (e11 or e12) is capable of emitting the signal required for m1 descendants to form comb plates. Previous cell lineage analyses indicate that the two e1 daughters generate the same suite of cell fates. On the other hand, the m1 daughters (m11 and m12) normally give rise to different cell fates. Reciprocal m1 daughter deletions show that in the absence of one daughter, the other cell can generate all the cell types normally formed by the missing cell. Together, these findings demonstrate that the two m1 daughters (m11 and m12) represent an embryonic equivalence group or field and that differences in the fates of the two m1 daughters are normally controlled by cell-cell interactions. These combined properties of ctenophore development, including the utilization of deterministic cleavage divisions, inductive interactions, and the establishment of embryonic fields or equivalence groups, are remarkably similar to those present in the development of various bilaterian metazoans. PMID- 15108815 TI - A mode of arthropod brain evolution suggested by Drosophila commissure development. AB - The evolutionary origin of the tritocerebral neuromere, which is a brain segment located at the junction between the supra- and subesophageal ganglia in most mandibulates (arthropods such as crustaceans and insects), is a subject rich in contentious debate. Various models have argued that the tritocerebrum came from a segmental nerve cord ganglia that was recruited into the head during the course of arthropod evolution. However, despite much thought on the subject, the origin of the tritocerebrum remains obscure. Here I describe the development of the tritocerebral commissure in Drosophila and demonstrate that the tritocerebral and mandibular commissures actually form as one commissure and then separate in a manner very similar to how the anterior and posterior commissures of a ventral nerve cord neuromere form. I propose that the tritocerebral neuromere originated from the splitting of an ancestral neuromere located in the anterior subesophageal ganglion into distinct tritocerebral and mandibular neuromeres. Also, I discuss the problem of arthropod brain neuromere homology in reference to this hypothesis. PMID- 15108816 TI - Neural crest cells provide species-specific patterning information in the developing branchial skeleton. AB - The skeletal elements of the branchial region are made by neural crest cells, following tissue interactions with the pharyngeal endoderm. Previous transplantation experiments have claimed that the cranial neural crest is morphogenetically prespecified in respect of its branchial skeletal derivatives, that is, that information for the number, size, shape, and position of its individual elements is already determined in these cells when they are still in the neural folds. This positional information would somehow be preserved during delamination from the neural tube and migration into the branchial arches, before being read out as a spatial pattern of chondrogenesis and osteogenesis. However, it now appears that signals from the endoderm are able to specify not only the histogenic differentiation state of neural crest cells but also the identity and orientation of the branchial skeletal elements. It is therefore important to ask whether fine details of branchial skeletal pattern such as those that exist between different species are also governed by extrinsic factors, such as the endoderm, or by the neural crest itself. We have grafted neural crest between duck and quail embryos and show that the shape and size of the resulting skeletal elements is donor derived. The ability to form species-specific patterns of craniofacial skeletal tissue thus appears to be an inherent property of the neural crest, expressed as species-specific responses to endodermal signals. PMID- 15108818 TI - Comparative and phylogenetic analysis of developmental sequences. AB - Event pairing has been proposed for the optimization of developmental sequences (event sequences) on a given phylogenetic hypothesis (cladogram) to determine instances of sequence heterochrony. Here, we show that event pairing is faulty, leading to the optimization of impossible hypothetical ancestors, the underestimation of the lengths of the developmental sequences on the tree, and the proposition of synapomorphies that are not supported by the data. When used for phylogenetic analysis, event pairing can even produce cladograms that are inconsistent with the data. These errors are caused by the fact that event pairing treats dependent features as if they were independent. We present a new method for comparative and phylogenetic analysis of developmental sequences that does not exhibit these errors. Our method applies Search-based character optimization and treats the entire developmental sequence as a single character that is then analyzed by using an edit cost function, which specifies the transformation cost between pairs of observed and unobserved character states, and dynamic programming. In other words, the developmental sequence is directly optimized on the tree. We used event pairing as an edit cost function, but others are possible. PMID- 15108817 TI - Enlightenment of old ideas from new investigations: more questions regarding the evolution of bacteriogenic light organs in squids. AB - Bioluminescence is widespread among many different types of marine organisms. Metazoans contain two types of luminescence production, bacteriogenic (symbiotic with bacteria) or autogenic, via the production of a luminous secretion or the intrinsic properties of luminous cells. Several species in two families of squids, the Loliginidae and the Sepiolidae (Mollusca: Cephalopoda) harbor bacteriogenic light organs that are found central in the mantle cavity. These light organs are exceptional in function, that is, the morphology and the complexity suggests that the organ has evolved to enhance and direct light emission from bacteria that are harbored inside. Although light organs are widespread among taxa within the Sepiolidae, the origin and development of this important feature is not well studied. We compared light organ morphology from several closely related taxa within the Sepiolidae and combined molecular phylogenetic data using four loci (nuclear ribosomal 28S rRNA and the mitochondrial cytochrome c oxidase subunit I and 12S and 16S rRNA) to determine whether this character was an ancestral trait repeatedly lost among both families or whether it evolved independently as an adaptation to the pelagic and benthic lifestyles. By comparing other closely related extant taxa that do not contain symbiotic light organs, we hypothesized that the ancestral state of sepiolid light organs most likely evolved from part of a separate accessory gland open to the environment that allowed colonization of bacteria to occur and further specialize in the eventual development of the modern light organ. PMID- 15108819 TI - Autopsy--the lost opportunity. PMID- 15108820 TI - The prozone phenomenon in syphilis testing. PMID- 15108821 TI - AIDS--no longer a death sentence, still a challenge. PMID- 15108822 TI - Primary esophageal lymphoma: a diagnostic challenge. PMID- 15108823 TI - GALOP syndrome: a treatable immune-mediated late-age onset polyneuropathy with gait ataxia. PMID- 15108824 TI - Consecutive autopsies on an internal medicine service. AB - OBJECTIVES: Autopsy rates continue to decline in the United States despite the demonstrated value of this procedure in many different settings. We sought to review clinical pathologic discordance information generated by autopsies on an internal medicine service in the urban United States and to determine whether resident services appear to influence autopsy rates. METHODS: We reviewed consecutive deaths and autopsies on an inpatient internal medicine service during a 30-month period at a 400-bed community hospital in Baltimore, MD. RESULTS: There were 622 deaths and 65 autopsies (10.3%). Resident teaching status correlated with a higher rate of autopsies performed (P = 0.048). Clinical pathologic discordance was common, with a major discordance rate of 39%. Major discordance was indicated by only one of nine autopsies performed on patients with human immunodeficiency virus. CONCLUSIONS: The autopsy was a valuable educational and quality improvement tool on the urban internal medicine service. Residency influences may be a major factor in continuing this exercise. In our study, although the numbers were small, patients with human immunodeficiency virus had a very low discordance rate. PMID- 15108825 TI - Treatment of HIV/AIDS in the nursing home: variations in rural and urban long term care settings. AB - OBJECTIVES: With the increased use of antiretroviral therapy, more patients with human immunodeficiency virus and acquired immunodeficiency syndrome (HIV/AIDS) are surviving for long periods of time. The aim of this study was to determine the availability of specialty HIV/AIDS services in long-term care facilities, and to determine differences in the availability of these services between rural and urban nursing homes. METHODS: 1,423 nursing homes from the 1999 National Nursing Home Survey were stratified by rural/urban status and compared using chi2 analysis and logistic regression. RESULTS: Less than 1% of surveyed nursing homes in the United States provided specialty HIV/AIDS services. While there was evidence that larger nursing homes are more likely to provide HIV/AIDS-related services, there were no significant differences between rural and urban nursing homes in the provision of specialty HIV/AIDS services. CONCLUSIONS: A vast majority of nursing homes in the United States do not provide any specialty areas for HIV/AIDS care. As our population ages and the life span of those diagnosed with HIV/AIDS continues to increase, nursing homes will begin to see patients diagnosed with HIV/ AIDS among those seeking care. PMID- 15108826 TI - Gender differences in knowledge and perceptions of HIV resources among individuals living with HIV in the Southeast. AB - OBJECTIVES: Ancillary services have been associated with beneficial health utilization outcomes among individuals infected with the human immunodeficiency virus (HIV), including greater retention in medical care and greater likelihood of antiretroviral use. Our primary objectives were to examine gender differences in barriers to ancillary services among people living with HIV in the Southeastern United States. METHODS: Survey and chart abstraction data were collected from six tertiary infectious diseases clinics in the Southeast. Using multivariate analyses, we examined the relationship between gender and 1) knowledge of how to access HIV and acquired immunodeficiency syndrome (AIDS) resource information and 2) opinions about the helpfulness of local services for people with HIV/AIDS. RESULTS: Women were less knowledgeable about HIV/AIDS resources and rated local services less favorably than men. Middle-aged and older African-American women rated local services as less helpful than other survey participants did. CONCLUSIONS: These findings indicate a need for outreach services that are designed to address the specific needs of older African American women, and women in general. PMID- 15108827 TI - Relationship between some acute phase reactants and the Bath Ankylosing Spondylitis Disease Activity Index in patients with ankylosing spondylitis. AB - OBJECTIVES: The aims of this study were to investigate a possible relationship between the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and some acute phase reactant (APR) levels in patients with ankylosing spondylitis (AS). METHODS: Twenty outpatients who fulfilled the modified New York criteria for AS were included in the study. Laboratory activity was assessed by examining erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), haptoglobin (Hp), and beta2 microglobulin (beta2MG). Disease activity was assessed according to the BASDAI, which includes a 10-point visual analogue scale to measure pain, fatigue, morning stiffness, swelling, and areas of local tenderness. RESULTS: When APR values were analyzed for the BASDAI, a positive correlation between CRP and BASDAI was observed (r = 0.556, P < 0.05). There was no clear, statistically significant correlation between BASDAI and the other APRs (ESR, r = 0.328, P > 0.05; Hp, r = 0.035, P > 0.05; and beta2MG, r = -0.190, P > 0.05). CONCLUSIONS: Our data suggest that CRP is a better marker of disease activity than ESR, Hp, and beta2MG. PMID- 15108828 TI - Dolichonychia in women with Marfan syndrome. AB - OBJECTIVES: Dolichonychia refers to nails that are elongated and slender. This nail morphology has been described in single case reports of patients with Ehlers Danlos syndrome, hypohidrotic ectodermal dysplasia, and Marfan syndrome. METHODS: The length and width of fingernails were measured in patients with Marfan syndrome and in individuals without the syndrome. A fingernail index was calculated for each person. A non-parametric sign test was used to compare the mean fingernail index of patients with Marfan syndrome and control subjects. RESULTS: The fingernail index ranged from 0.92 to 1.52 in patients with Marfan syndrome and from 0.77 to 1.30 in control individuals. The median fingernail index in women with Marfan syndrome (1.455) was significantly greater than the median fingernail index in women control subjects (1.080) (P = 0.035). Dolichonychia, defined as a fingernail index greater than or equal to 1.30, was present in six of eight (75%) of women with Marfan syndrome as compared with only one of 40 (2.5%) of women control subjects. CONCLUSIONS: Dolichonychia is a physical finding present in some patients with Marfan syndrome. The detection of long narrow fingernails in women should suggest the possibility of Marfan syndrome. PMID- 15108829 TI - Incidence of postdural puncture headache and backache, and success rate of dural puncture: comparison of two spinal needle designs. AB - OBJECTIVES: The goal of this randomized study was to compare the incidence of postdural puncture headache (PDPH) and postdural puncture backache (PDPB), and the success rate between two small-gauge spinal needle designs used in women undergoing subarachnoid block anesthesia. METHODS: After Institutional Review Board approval, 215 patients presenting for tubal ligation were randomly assigned to have 26-gauge Atraucan (AT group) or 25-gauge Whitacre (WH group) spinal needles used in their spinal anesthesia. The number of attempts to successful cerebrospinal fluid return and the success rate of the spinal blockade were documented. Postoperatively, an investigator blinded to the study interviewed patients daily. RESULTS: The incidence of PDPH was similar between the AT group (3.9%) and the WH group (4.0%). The total duration of all PDPHs was 5 days for the AT group and 15 days for the WH group. Both groups had a similar one-attempt success rate of 61% (AT group) and 62% (WH group). Failure to obtain cerebrospinal fluid occurred in only one in the AT group and two in the WH group. The incidence of PDPB was similar and the severity was mild in both groups. CONCLUSIONS: The low complication and failure rates make these two types of smaller size spinal needle design good candidates for dural puncture procedures, such as spinal anesthesia, diagnostic lumbar punctures, and myelograms. PMID- 15108830 TI - Hepatitis C infection: a clinical review. AB - Nearly three million persons in the United States are viremic with hepatitis C (HCV). Despite a decreasing incidence of HCV in this country, the prevalence of HCV-related chronic liver disease is increasing. Most infections in the United States are acquired by intravenous drug use. The chronicity rate of HCV is high, reaching 85% in some populations, and the risk of progression to advanced liver disease is as high as 20% within twenty years of infection. Host factors like alcohol use accelerate the rate of progression. The enzyme immunoassay is the preferred initial test for diagnosis; the third generation assay has greater than a 99% specificity in immunocompetent patients. Barring contraindications, the standard of care for treatment of chronic HCV has become pegylated interferon and ribavirin. With this therapy, the cure rate for treatment-naive patients is about 55%, but rates are higher in certain groups. Common side effects of therapy include neuropsychiatric symptoms, influenza-like symptoms and hematological abnormalities. PMID- 15108831 TI - Smallpox vaccination in the early 19th century using live carriers: the travels of Francisco Xavier de Balmis. AB - Realizing that the Spanish colonies were being devastated by epidemics of smallpox resulting in thousands of deaths, Charles IV, King of Spain, sent one of his court's physicians to apply the recently discovered vaccine. Without refrigeration, the vaccine was passed from one child to another (boys taken out of orphanages). Francisco Xavier de Balmis and a team that included three assistants, two surgeons, and three nurses sailed from Spain on November 30. 1803. They vaccinated more than 100,000 people from the Caribbean Islands and South, Central, and North America, reaching up to San Antonio, Texas, and then traveled to the Philippines, Macao, Canton, and Santa Elena Island, landing back in Cadiz on September 7, 1806. During his journey, Balmis instructed local physicians on how to prepare, preserve, and apply the vaccine, while collecting rare biologic specimens. On the 200th anniversary of their sailing, recognition is given to this group for conducting what was the first global vaccination campaign that reached Texas and California. PMID- 15108832 TI - The prozone phenomenon with syphilis and HIV-1 co-infection. AB - The prozone phenomenon in syphilis testing refers to a false negative response resulting from overwhelming antibody titers which interfere with the proper formation of the antigen-antibody lattice network necessary to visualize a positive flocculation test. This prozone effect in syphilis testing can be expected in cases of disproportionately high antibody titers, such as secondary syphilis, or with human immunodeficiency virus (HIV) coinfection. Clinicians need to remain familiar with the protean manifestations of syphilis to be able to exclude the prozone phenomenon. PMID- 15108833 TI - Primary esophageal lymphoma: a diagnostic challenge in acquired immunodeficiency syndrome--two case reports and review. AB - Although extranodal presentation occurs in the majority of cases of acquired immunodeficiency syndrome-associated non-Hodgkin lymphoma, the esophagus is only rarely affected. We discuss two patients with acquired immunodeficiency syndrome who presented with dysphagia and weight loss, who were found to have human immunodeficiency virus-associated primary esophageal lymphoma. Both patients died within a few weeks of diagnosis, reflecting the poor prognosis associated with this malignancy. Primary esophageal lymphoma should be considered in the differential diagnosis in a human immunodeficiency virus-seropositive patient presenting with dysphagia. PMID- 15108834 TI - Leukemoid reaction due to Clostridium dificile infection in acquired immunodeficiency syndrome: two case reports and a review of the literature. AB - The clinical presentation of colitis associated with Clostridium difficile infection in immunosuppressed patients with acquired immunodeficiency syndrome (AIDS) has not been completely characterized. Previous reports suggest that these patients present with low blood leukocyte counts, consistent with the impaired myelopoiesis that can occur with human immunodeficiency virus (HIV) infection. In contrast, we describe the cases of two patients with colitis associated with C difficile infection who developed intense leukemoid reactions despite being in advanced stages of AIDS. To the best of our knowledge, these are the first described cases of leukemoid reaction associated with C difficile or other bacterial infection in AIDS patients. We review the literature on C difficile colitis in patients infected with HIV and suggest that severe C difficile infection should be considered in such patients presenting with leukemoid reaction and diarrhea. PMID- 15108835 TI - Primary human immunodeficiency virus infection presenting as acute pancreatitis. AB - Primary human immunodeficiency virus (HIV) infection can present in a variety of ways. It is important to recognize acute HIV infection, for personal and public health reasons. We present an unusual case of primary HIV infection manifesting as acute pancreatitis. PMID- 15108836 TI - Corynebacterium afermentans lung abscess and empyema in a patient with human immunodeficiency virus infection. AB - Necrotizing pleuropulmonary infection in a patient with acquired immunodeficiency syndrome developed due to Corynebacterium afermentans subspecies lipophilum. Long term combination antibiotic therapy was successful in eradicating the infection without surgery. PMID- 15108837 TI - Histoplasmosis in two human immunodeficiency virus-positive immigrants to Italy: clinical features and management in the highly active antiretroviral therapy era. AB - We report two cases of histoplasmosis occurring in human immunodeficiency virus positive patients who immigrated to Italy, and focus our attention on the clinical features and therapeutic aspects, with particular emphasis on secondary prophylaxis. The patients had comparable human immunodeficiency virus baseline parameters, but had a completely different compliance over therapeutic regimens. The two patients were followed in two different city hospitals of our region, Padua and Verona, and the diagnosis was made on the basis of instrumental, histologic, and microbiologic findings. One of them was treated with corticosteroids because of nephrotic syndrome. PMID- 15108838 TI - Hepatocellular carcinoma in a patient with human immunodeficiency virus and hepatitis B virus coinfection: an emerging problem? AB - Infections with hepatitis B virus (HBV) and human immunodeficiency virus (HIV) have similar risk factors and routes of transmission. It is estimated that 64 to 84% of HIV-infected individuals have positive markers for anti-HBc antibodies, with the chronic HBV infection rate approaching 16%. There is, however, a paucity of information on HBV/HIV coinfection, and its clinical implications remain unclear. We review the literature and report our recent experience with a 44-year old man with HBV/HIV coinfection who developed metastatic hepatocellular carcinoma despite quiescent HBV and HIV disease courses. Highly active antiretroviral therapy has revolutionized HIV disease. As a result, morbidity and mortality from other underlying chronic, non-HIV-related diseases, such as the HBV infection and hepatocellular carcinoma reported here, will likely continue to increase in the HIV-infected patient population. PMID- 15108839 TI - Late failure of combined recombinant hepatitis B vaccine and lamivudine in treatment of a patient with chronic hepatitis B. AB - We report the first case of a woman having chronic hepatitis B treated with a combination therapy of recombinant hepatitis B vaccine and lamivudine for 18 months. The main aims of such a combined therapy were to assess whether the concomitant anti-hepatitis B virus (HBV) vaccination might prevent the emergence of a mutant HBV and lead to sustained hepatitis B e antigen seroconversion with undetectable serum HBV DNA. The data from the present case demonstrated that combination of anti-HBV vaccine and lamivudine did not eliminate viral DNA despite prolonged treatment and did not have any effect on preventing resistant type HBV. Although the combined therapy failed to reach the therapeutic endpoints, it concerned a single and unique patient. Hepatitis B vaccine and lamivudine for HBV treatment should be further investigated in randomized controlled trials. PMID- 15108840 TI - GALOP syndrome: case report with 7-year follow-up. AB - An elderly woman complaining of a gait disorder was found to have the GALOP syndrome (gait ataxia, late-onset polyneuropathy). She exhibited mild distal weakness and sensory loss in the legs, a positive Romberg, and an unsteady gait. Serum immunofixation disclosed a monoclonal IgM-kappa protein. There was specific IgM binding to galopin, a central nervous system white matter antigen. Periodic treatment with intravenous immunoglobulin has alleviated her neurologic symptoms. She has now been followed for 7 years and maintained significant improvement in neurologic symptoms and signs. PMID- 15108841 TI - Acute splenic sequestration crisis resembling sepsis in an adult with hemoglobin SC disease. AB - Acute splenic sequestration crisis (ASSC) is a common complication of sickle cell anemia in children. ASSC is generally not seen in adults with the SS genotype but occasionally can be seen in adults with the SC genotype. We present a case of fulminant ASSC in an adult with hemoglobin SC who developed high fever, intense abdominal pain, leukocytosis, and jaundice. PMID- 15108842 TI - Otogenic Fusobacterium meningitis, sepsis, and mastoiditis in an adolescent. AB - Fusobacterium necrophorum is a strict anaerobic organism responsible for a number of clinical syndromes known as necrobacillosis. Although meningeal infections with anaerobes are rare, delayed diagnosis and treatment can be potentially fatal. We report a unique case of Fusobacterium meningitis, mastoiditis, and sepsis in a previously healthy adolescent. Diagnosis and management of this condition are discussed in the context of a literature review. PMID- 15108843 TI - Coccidioidomycosis of the prostate gland: two cases and a review of the literature. AB - Coccidioidomycosis prostatitis is an uncommon presentation of disseminated coccidioidomycosis, a fungal disease endemic in the southwestern United States. Coccidioidomycosis prostatitis should be considered in the differential diagnosis of a patient from an endemic region with evidence of persistent sterile pyuria, prostatitis, or granulomatous disease of the prostate. Diagnosis is established by biopsy, and treatment includes either an azole or amphotericin B. We present the twelfth and thirteenth reported cases and provide a review of the literature. PMID- 15108844 TI - False-positive human immunodeficiency virus test and Trypanosoma cruzi infection in eastern Colombia. PMID- 15108845 TI - What goes around, comes around. PMID- 15108846 TI - NIH asks participants in Women's Health Initiative estrogen-alone study to stop study pills, begin follow-up phase. PMID- 15108847 TI - [Nothing is more damaging to new truth than an old misconception. Goethe, Wilhelm Meisters Wanderjahre]. PMID- 15108848 TI - [Refreshing in abdominal ultrasonography--a summary from SGUM Kongress Davos in 2003]. AB - Ultrasonography is a modern noninvasive imaging technique with a high resolution, and high evidence in diagnostic questions, high acceptance in patients, low costs and no side effects. There are pure questions in whitch the diagnostic ultrasound is not able to give a satisfactory answer. Important is basically a well trained examinator with long experience and an exact examination and documentation. PMID- 15108849 TI - [Hip sonography for newborn screening]. AB - In the past, congenital dysplasia and dislocation of the hip joint have, despite treatment, led frequently to late sequals (deformations and incongruous articular surfaces), requiring repeated corrective interventions. Nevertheless, life-long handicaps could not always be prevented. It is known for a long time that sequels, such as articular deformations and desorders of growth of the femoral head, can be prevented the better, the earlier the treatment is initiated. However, a reliable method for early diagnosis has been lacking to date. Even systematic clinical examinations of all newborn babies have not been able to prevent late appearance of dysplasia and dislocation. This is not due to poor practice of clinical examination, but is explained by to day's knowledge that most dislocations are not present at birth but develop later on. Deformations of the acetabular roof without any dislocation of the femoral head are clinically not papable and do not cause any symptoms in adults until the onset of coxarthritis. Hip sonography according to Graf has proven to be an effective imaging procedure for early diagnosis. Right at birth, it allows very reliable detection of all cases that, if left untreated, will cause later major therapeutic difficulties, such as dislocations of the femoral head or coxarthritis due to dysplasia of the acetabular roof. If diagnosed early--at best immediately after birth--successful healing can be achieved even in severe cases resulting in an anatomical and functional normal hip joint, only by conservative and out-patient treatment. That is why many centers have established sonographic screening programs of all newborns as a preventive measure. Screening is medically most effective, if practiced at the earliest possible stage (at best immediately after birth). From an economical point of view, no disadvantage of early screening (despite of higher rates of findings requiring review) has been found in terms of overall costs (screening, follow-up, treatment). PMID- 15108850 TI - [Thoraxsonography--Part 1: Chest wall and pleura]. AB - The detection of lymph nodes and a careful interpretation of dignity is one of the main indications for chest wall sonography. Palpable unclear masses of the chest wall can undergo US guided puctures very easily. Either rib and sternum fractures are detectable very sensitively. In addition to the more sensitive detection than in normal X-ray also accompanying soft parts processes, pleural effusions and haematomas can be visualised. Despite the physical laws of ultrasound, approximately 70% of the pleural surface can be accessed by sonography. The normal parietal pleural can be visualized and delineated from circumscribed as well as diffuse pathological thickening. The normal visceral pleura is hidden by the total reflection at the surface of the aerated lung. Sonographic evidence of a pleural effusion can be obtained from 5 ml onwards and is much more sensitive than a chest X-ray, especially in supine position; false positive findings are not encountered. A more accurate estimation of the quantity of effusion can be made. Exudates are echogenic in one third of cases. Pleural thickening, nodular changes in the pleura, septa, and the formation of the quality of effusion can be made. Exual exudates, whereas transudates are always anechoic. Pleural metastases are characteristically hypoechoic and nodular polypoid. Pleural mesotheliomas can be delineated nearly equally well on sonograms and on computed tomography. Morphological and, in particular, functional diagnosis of the diaphragm by means of sonography is both reliable and convincing. PMID- 15108851 TI - ["Antibiotic stewardship": catchword or necessity?]. PMID- 15108852 TI - [Fish oil for the psyche?]. PMID- 15108853 TI - The future of biotechnology in support of bio-based industries: the US perspective. PMID- 15108854 TI - Land use-related chemical composition of street sediments in Beijing. AB - BACKGROUND: More than 10 million people are currently living in Beijing. This city faces severe anthropogenic air pollution caused by an intense vehicle increase (11% per year in China), coal combusting power plants, heavy industry, huge numbers of household and restaurant cookers, and domestic heating stoves. Additionally, each year dust storms are carrying particulate matter from the deserts of Gobi and Takla Makan towards Beijing, especially in spring. Other geogenic sources of particulate matter which contribute to the air pollution are bare soils, coal heaps and construction sites occurring in and around Beijing. Streets function as receptor surfaces for atmospheric dusts. Thus, street sediments consist of particles of different chemical compositions from many different sources, such as traffic, road side soils and industry. METHODS: Distributions and concentrations of various chemical elements in street sediments were investigated along a rural-urban transect in Beijing, China. Chemical elements were determined with X-ray fluorescence analysis. Factor analysis was used to extract most important element sources contributing to particulate pollution along a main arterial route of the Chinese capital. RESULTS AND DISCUSSION: The statistical evaluation of the data by factor analysis identifies three main anthropogenic sources responsible for the contamination of Beijing street sediments. The first source is a steel factory in the western part of Beijing. From this source, Mn, Fe, and Ti were emitted into the atmosphere through chimneys and by wind from coal heaps used as the primary energy source for the factory. The second source is a combination of traffic, domestic heating and some small factories in the center of Beijing discharging Cu, Pb, Zn and Sn. Calcium and Cr characterize a third anthropogenic element source of construction materials such as concrete and mortar. Beside the anthropogenic contamination, some elements like Y, Zr, Nb, Ce, and Rb are mainly derived from natural soils and from the deserts. This is supported by mineral phase analysis, which showed a clear imprint of material in road dusts coming from the West-China deserts. CONCLUSIONS: Our results clearly show that the chemical composition of urban road dusts can be used to identify distinct sources responsible for their contamination. The study demonstrates that the chemistry of road dusts is an important monitor to assess the contamination in the urban environment. Chemical composition of street sediments in Beijing comprises the information of different sources of atmospheric particles. RECOMMENDATIONS AND OUTLOOK: This study is only a small contribution to the understanding of substance fluxes related to Beijing's dust. More effort is required to assess Beijing's dust fluxes, since the dust harms the living quality of the inhabitants. Especially the measurable superimposing of long scale transported dust from dry regions with the anthropogenic polluted urban dust makes investigations of Beijing's dust scientifically valuable. PMID- 15108855 TI - Short-term model of the production of construction aggregates in Taiwan based on artificial neural networks. AB - BACKGROUND: Taiwan's geography and limited stock of sandstone have caused sandstone resources to gradually decline to the point of exhaustion after long term excavation. Moreover, the Taiwanese government has continuously increased the amount of land area near rivers that cannot be excavated to facilitate riverbed remediation and promote conservation of water resources. Accordingly, predicting and managing the annual production of construction aggregates in future construction projects, and dealing appropriately with some thorny problems, for instance, demand that excess supply, excessive excavation, unregulated excavation, and the consequent environmental damage, will significantly affect the efficient use of natural resources in a manner that accords with the national policy of Sustainable Development (SD). METHODS: . This study establishes an empirical model for forecasting the annual production of future construction aggregates using Artificial Neural Networks (ANN), based on 15 relevant socio-economic indicators, such as indicator of annual consumption of cement. A sensitivity analysis is then performed on these indicators. RESULTS AND DISCUSSION: This work applies ANN to estimate the annual production of construction aggregates; the estimates, the verification of the model and the sensitivity analysis are all acceptable. Furthermore, sensitivity analysis results indicate that the annual consumption of cement is the indicator that most strongly influences the production of construction aggregates, as well as whether construction waste can be recycled and steel structures can be used in buildings, helping to reduce the future production of construction aggregates in Taiwan. CONCLUSIONS: The elaborate prediction methodology presented in this study avoids some of the weaknesses or limitations of conventional linear statistics, linear programming or system dynamics. Additionally, the results not only provide a short-term prediction of the production of construction aggregates in Taiwan, but also provide a viable and flexible means of verifying quality certification of the production data of construction aggregates in the future by incorporating those relevant socio-economic indicators. RECOMMENDATIONS AND OUTLOOK: The continuity and quality of the database of relevant indicators used in this study should be closely scrutinized in order to ensure the SD means of exploiting resources. PMID- 15108856 TI - (M)VOC and composting facilities. Part 1: (M)VOC emissions from municipal biowaste and plant refuse. AB - GOAL, SCOPE AND BACKGROUND: Malodorous volatiles derived from the decomposition of biowaste within the process of composting might pose a risk to human health. Different techniques of process engineering have been developed to minimise the burden of malodorous compounds in air possibly affecting compost workers and residents in the vicinity. METHODS: In the present study, three different composting facilities were examined for the emission of volatiles to estimate the impact of process engineering on the dispersal of odorous compounds and to discuss its relevance for human health. RESULTS AND DISCUSSION: Concentrations of single compounds belonging to alcohols, ketones, furanes, sulfur-containing compounds and especially terpenes ranged from 10(2) up to nearly 10(6) ng/m3 depending on the sampling sites and the process engineering. The ratio of MVOC and total VOC measured changed throughout the process of biodegradation. A certain combination of volatile compounds coincided with the occurrence of typical compost odour. CONCLUSION: The type of process engineering seemed to have a major impact on the emission of volatiles, as amounts of (microbial) volatiles emitted were characteristic for the different techniques used. Thus, the MVOC emission basically depends on the degree of biodegradation. It is likely that the concentrations workers are exposed to can have an impact on human health. RECOMMENDATIONS AND OUTLOOK: It is obvious that less sophisticated types of process engineering give rise to greater amounts of bioaerosols and volatiles and, therefore, technical devices have to be improved and controlled regularly to minimise adverse health effects on workers. PMID- 15108857 TI - Effects of lindane on the photosynthetic apparatus of the cyanobacterium Anabaena: fluorescence induction studies and immunolocalization of ferredoxin NADP+ reductase. AB - INTENTION, GOAL, SCOPE, BACKGROUND: Cyanobacteria have the natural ability to degrade moderate amounts of organic pollutants. However, when pollutant concentration exceeds the level of tolerance, bleaching of the cells and death occur within 24 hours. Under stress conditions, cyanobacterial response includes the short-term adaptation of the photosynthetic apparatus to light quality, named state transitions. Moreover, prolonged stresses produce changes in the functional organization of phycobilisomes and in the core-complexes of both photosystems, which can result in large changes in the PS II fluorescence yield. The localization of ferredoxin-NADP+ reductase (FNR) at the ends of some peripheral rods of the cyanobacterial phycobilisomes, makes this protein a useful marker to check phycobilisome integrity. OBJECTIVE: The goal of this work is to improve the knowledge of the mechanism of action of a very potent pesticide, lindane (gamma hexaclorociclohexane), in the cyanobacterium Anabaena sp., which can be considered a potential candidate for bioremediation of pesticides. We have studied the effect of lindane on the photosynthetic apparatus of Anabaena using fluorescence induction studies. As ferredoxin-NADP+ reductase plays a key role in the response to oxidative stress in several systems, changes in synthesis, degradation and activity of FNR were analyzed. Immunolocalization of this enzyme was used as a marker of phycobilisome integrity. The knowledge of the changes caused by lindane in the photosynthetic apparatus is essential for rational further design of genetically-modified cyanobacteria with improved biorremediation abilities. METHODS: Polyphasic chlorophyll a fluorescence rise measurements (OJIP) have been used to evaluate the vitality and stress adaptation of the nitrogen-fixing cyanobacterium Anabaena PCC 7119 in the presence of increasing concentrations of lindane. Effects of the pesticide on the ultrastructure have been investigated by electron microscopy, and FNR has been used as a marker of phycobilisome integrity. RESULTS AND DISCUSSION: Cultures of Anabaena sp. treated with moderate amounts of lindane showed a decrease in growth rate followed by a recovery after 72 hours of pesticide treatment. Concentrations of lindane below 5 ppm increased the photosynthetic performance and activity of the cells. Higher amounts of pesticide caused a decrease in these activities which seems to be due to a non-competitive inhibition of PS II. Active PS II units are converted into non-QA reducing, so called heat sink centers. Specific activity and amount of FNR in lindane-treated cells were similar to the values measured in control cultures. Release of FNR from the thylakoid after 48 hours of exposure to 5 ppm of lindane towards the cytoplasm was detected by immunogold labeling and electron microscopy. CONCLUSIONS: From these results, we conclude that the photosynthetic performance and activity of the cells are slightly increased in the presence of lindane up to 5 ppm. Moreover, in those conditions, lindane did not produce significant changes in the synthesis, degradation or activity of FNR. The high capability of Anabaena to tolerate lindane makes this cyanobacterium a good candidate for phytoremediation of polluted areas. RECOMMENDATION AND OUTLOOK: The results of this study show that cultures of Anabaena PCC 7119 tolerate lindane up to 5 ppm, without significant changes in the photosynthetic vitality index of the cells. However, a slight increase in phycobiliprotein synthesis is observed, which is related to total protein content. This change might be due to degradation of proteins less stable than phycobiliproteins. An identification of the proteins with altered expression pattern in the presence of the pesticide remains the subject of further work and will provide valuable information for the preparation of strains which are highly tolerant to lindane. PMID- 15108858 TI - Photodegradation and volatility of pesticides: chamber experiments. AB - BACKGROUND AND OBJECTIVES: Among the factors affecting the environmental fate of surface-applied pesticides several biological as well as abiotic factors, such as volatilization and photochemical transformations are of particular interest. Whereas reliable measurement methods and models for estimating direct photodegradation are already available for the compartments of water and atmosphere and individual subprocesses have already been described in detail, there is still a need for further elucidation concerning the key processes of heterogeneous photodegradation of environmental chemicals on surfaces. METHODS: In order to systematically examine the direct and indirect photodegradation of 14C-labeled pesticides on various surfaces and their volatilization behavior, a new laboratory device ('photovolatility chamber') was designed according to US EPA Guideline 161-3. Model experiments under controlled conditions were conducted investigating the impact of different surfaces, i.e. glass, soil dust and radish plants, and environmental factors, i.e. irradiation and atmospheric ozone (O3), on the photodegradation and volatilization of surface-deposited [phenyl-UL 14C]parathion-methyl (PM). RESULTS AND DISCUSSION: Depending on the experimental conditions, parathion-methyl was converted to paraoxon-methyl, 4-nitrophenol, unknown polar products and 14CO2. With respect to the direct photodegradation of PM (experiments without O3), the major products were polar compounds and 14CO2, due to the rapid photochemical mineralization of 4-nitrophenol to 14CO2. Paraoxon methyl and 4-nitrophenol formation was mainly mediated by the combination of light, O3, and *OH radicals. In radish experiments PM photodegradation was presumably located in the cuticle compartment, which exhibited a sensitized photodegradation, as more unknown products were yielded compared to the glass and soil dust experiments. This could be explained by intensifying the inherent PM degradation in the dark with the same product spectrum. Due to photochemical product formation, which is an antagonistic process to the volatilization of parent compound, the volatilization of unaltered parathion-methyl from each surface generally decreased in the presence of light, particularly in combination with increasing O3 concentrations and *OH radical production rates. CONCLUSION: First results demonstrated that the photovolatility chamber provides a special tool for the systematic evaluation of (a) photodegradation of surface-located pesticide residues, i.e. measuring qualitative aspects of direct and indirect photodegradation together with relative photodegradation rates, and (b) volatilization of pesticides on surfaces by including and optionally varying relevant parameters such as light, atmospheric O3 concentration, surface temperature, air temperature, air flow rate. OUTLOOK: The experimental facility represents an important complement to lysimeter and field studies, in particular for experiments on the volatilization of pesticides using the wind tunnel system. With the photovolatility chamber special experiments on photodegradation, volatilization and plant uptake can be conducted to study key processes in more detail and this will lead to a better understanding of the effects of certain environmental processes on the fate of released agrochemicals contributing to an improved risk assessment. PMID- 15108859 TI - The effect of 4-nonylphenol on the pigmentation of Ocimum basilicum (Basil). AB - BACKGROUND, AIM AND SCOPE: Tests during the last few years have confirmed that 4 nonylphenol (4-NP) can have oestrogen-like effects (xeno-hormone) on animal organisms. The objective was to firstly evaluate the ecotoxicological effects of 4-NP on plants such as hydrocultures. To clarify how this substance interferes with the photosynthetic system of plants, various tests were carried out using the basil plant (Ocimum basilicum). METHODS: The effect of the pollutant 4-NP on the pigment content in the leaves of the basil plant was analysed with the use of High-Performance-Liquid-Chromatography (HPLC). RESULTS AND DISCUSSION: A general assessment of the HPLC data revealed that plants that came in contact with the 4 nonylphenol showed a change in pigmentation. More chlorophyll a and b was produced, although at the same time a higher production of degradation products and by-products of the chlorophylls was observed. These occurrences can therefore be seen as an impairment of the photosynthetic process. The contaminated plants produced less xanthophylls than the non-contaminated ones, though these differences were statistically not significant. CONCLUSION AND PERSPECTIVE: The variations on the pigment content in the leaves of the basil plant can be interpreted as a consequence of the 4-NP application. It was, however, not investigated whether the plants absorbed the pollutants directly. The effect could have been caused by adsorption of the oily substance to the roots, and this could have led to a hindrance of the uptake of nutrients and possibly water. In order to clarify this further, biochemical experiments are being conducted. PMID- 15108860 TI - Topics of water sciences and technology. AB - In the preceding series of contributions to ESPR typical examples from actual fields of water chemistry were presented on various topics: integrated water quality management, diagnosis of water bodies, therapy of aquatic systems, and fitness for aquatic systems. These contributions clearly showed the need and importance of more intensive research. In the principle committee III 'Basic Research' of the Water Chemical Society, various expert groups work on scientific backgrounds in these fields. Some relevant topics are selected in this outlook, mainly on interactions of solids with water, the role of environmental colloids, the development of modern biochemical methods for diagnosis, the impact of chemical speciation on bioavailability and water technology, and the development of advanced methods in water treatment technology. Innovative approaches to understand the interactions between pollutants, water and solids are crucial for assessment of contaminants with biochemical and analytical methods, for the development of new efficient technologies and for application of treatment methods with little or no waste and by-product formation. PMID- 15108861 TI - The EULIMNOS project: strengthening cross-border scientific research and higher education in the Lake Shkodra/Skadar Region through a multidisciplinary transboundary approach to conservation. PMID- 15108862 TI - RECETOX (Brno, Czech Republic)--history, aims and activities. PMID- 15108863 TI - The efficacy of vaginal clindamycin for the treatment of abnormal genital tract flora in pregnancy. AB - OBJECTIVE: To assess the efficacy of 2% clindamycin vaginal cream (CVC) to treat bacterial vaginosis (BV) in pregnancy. METHODS: A prospective, randomized, double blind, placebo-controlled, tricenter study. Four hundred and four women with BV on Gram stain at their first antenatal clinic visit were randomized to receive a 3-day course of 2% CVC or placebo. The outcome was assessed using an intention to treat analysis at 3 weeks and 6 weeks post-treatment according to three different diagnostic methods based on five criteria (Gram stain and all four elements of clinical composite criteria: vaginal discharge, abnormal vaginal pH, clue cells, amine odor), three criteria (vaginal pH, clue cells, amine odor) or two criteria (clue cells and amine odor) to reflect stringency of diagnosis, historical precedence and government agency recommendations respectively. RESULTS: Using five diagnostic criteria, 18% of CVC patients were cured and 70.8% either cured and/or improved compared to 1.6% and 12% of placebo patients respectively (p < 0.0001). Using three diagnostic criteria, 44.8% of CVC patients were cured and 77.3% were either cured and/or improved compared to 9.3% and 28.8% of placebo patients respectively (p < 0.000 1). Using two diagnostic criteria, 75.0% of CVC patients were cured compared to 18.0% of placebo patients (p < 0.0001 ). Recurrence rates in those CVC patients successfully treated were approximately 6% at 6 weeks post baseline and 10% at 28 to 34 weeks gestation. CONCLUSIONS: A 3 day course of CVC appears to be well tolerated by the mother and statistically significantly more efficacious than placebo in the treatment of BV during the second trimester of pregnancy. PMID- 15108864 TI - Risk factors for hepatitis C infection among sexually transmitted disease infected, inner city obstetric patients. AB - OBJECTIVE: To test the hypothesis that our inner city obstetric patients who have been infected with sexually transmitted diseases (STDs) will have a higher prevalence of hepatitis C virus infection than the general population and to identify specific risk factors and high-risk groups. METHODS: All patients in our prenatal clinic (July 1997-April 1999) who tested positive for one or more STDs were asked to return for hepatitis C antibody testing. Medical charts of all patients who returned for hepatitis C testing were reviewed. RESULTS: A total of 106 patients with STDs were tested for hepatitis C. Positive screening tests for anti-hepatitis C antibody were found in 6.6% (7/106) of the patients (95% CI = 2.7-13.1%). This frequency is significantly higher than the hepatitis C prevalence (1.8%) in the general United States population (p = 0.006). Multiple logistic regression analysis confirmed only older age (p = 0.0 16) and positive HIV status (p = 0.023) to be significant predictors of hepatitis C infection. CONCLUSIONS: Inner city STD-infected obstetric patients are at high risk for hepatitis C infection compared with the general population. Increasing age and HIV-positive status are risk factors which are significantly associated with hepatitis C infection. PMID- 15108865 TI - Screening protocols for group B streptococcus: are transport media appropriate? AB - OBJECTIVE: To evaluate group B streptococcus (GBS) detection in an in vitro setting, using a low and controlled inoculum from swabs directly inoculated into a selective medium, as compared to delayed inoculation following a period in a commercial Amies transport medium with charcoal (Venturi Transystem Copan, Italy). STUDY DESIGN: Clinical isolates of GBS (n = 103), were inoculated into the Amies transport medium with charcoal in a concentration of 100 colony-forming units (cfu)/ml (10 cfu/swab). Swabs were then transferred to an enrichment broth (NPC) at time intervals of 0, 2, 4, 6 and 24 hours. Broths were then incubated for 18-24 hours at 35 degrees C in air, before being transferred to New Granada Medium Modified (NGM) for GBS detection and incubated for a further 18-24 hours at 35 degrees C in air. If the characteristic orange pigmented colonies were observed after this period, the specimen was recorded as + (1-10 colonies) or + + (more than 10 colonies). RESULTS: Overall 92.2% (95/103) of isolates were detected in all tubes and at all times. An additional two isolates were non hemolytic, non-pigment forming GBS. Of note, 3.9% (4/103) were negative until 2 hours delayed inoculation and 1.9% (2/103) gave inconsistent results, likely due to the low inoculum used. CONCLUSION: Delayed inoculation into selective enrichment broth following a period in transport medium, even with a low inoculum, gave a similar and acceptable GBS detection rate to direct inoculation. Hence, Amies transport medium with charcoal is an appropriate transport medium to use, where it is not practical for clinical specimens to be directly inoculated into selective enrichment broth and as endorsed in the Centers for Diseases Control (CDC) Guidelines, 2002. PMID- 15108866 TI - Maternal group B streptococcal (GBS) genital tract colonization at term in women who have asymptomatic GBS bacteriuria. AB - OBJECTIVE: To determine the rate of positive group B streptococcus (GBS) cultures at 35-37 weeks gestation in women who have first trimester asymptomatic GBS bacteriuria. METHODS: Pregnant women with asymptomatic first trimester GBS bacteriuria had genital cultures for GBS performed at 35-37 weeks gestational age. Serotyping was performed by the standard Lancefield capillary precipitin method. RESULTS: Fifty-three women with positive urine cultures had genital cultures performed at 35-37 weeks. Sixteen of the 53 (30.2%; 95% confidence interval: 18.4-44.3%) third trimester vaginal cultures were positive for GBS. Five of eight (63%) of the women with typable urine serotypes had the same typable serotype in the third trimester genital culture. CONCLUSION: Genital tract cultures at 35-37 weeks for GBS correlate poorly with first trimester asymptomatic GBS bacteriuria. Recommendations for GBS prophylaxis in labor in women who have first trimester asymptomatic GBS bacteriuria should be investigated further and reconsidered. PMID- 15108867 TI - Improving appropriate use of antifungal medications: the role of an over-the counter vaginal pH self-test device. AB - OBJECTIVES: To determine whether patients can understand and use the vaginal pH device in the diagnosis of vaginitis. To compare whether vaginal pH readings determined by patients and healthcare providers are similar. To determine whether vaginal pH can reduce inappropriate over-the-counter (OTC) antifungal medication use and improve the correct diagnosis of vaginitis. METHODS: One hundred and fifty-one women indicated their belief about the cause of their vaginal infection, read the instructions of the vaginal pH device package insert, used the device and interpreted the findings. The patient interpretations were compared with results obtained by healthcare providers, blinded to patient findings. RESULTS: Over 96% of patients stated that they could easily read the instructions, use the vaginal pH device and interpret the readings. They obtained the same readings as healthcare professionals (Kappa = 0.9). Restricting the use of OTC antifungal medications to those individuals with vaginitis symptoms and vaginal pH < or = 4.5 significantly reduced inappropriate use by approximately 50%, Fisher's exact test, p-value = 0.018. Conversely, seeking healthcare provider assessment with vaginal pH > 4.5, leads to correct diagnosis of vaginitis. CONCLUSIONS: The vaginal pH device can be used as an OTC diagnostic tool by consumers when a vaginal infection is suspected. Vaginal pH readings would direct patients whether to purchase an antifungal medication or seek professional diagnosis from a healthcare provider. Understanding and use of this vaginal pH device could reduce inappropriate use of OTC antifungal medications by approximately 50% and improve the correct diagnosis of vaginitis. PMID- 15108868 TI - Unilateral ovarian abscess caused by Salmonella. AB - BACKGROUND: Patients with unilateral ovarian abscesses due to Salmonella are rare. CASE REPORT: A 48-year-old woman with a left ovarian abscess caused by Salmonella group 07 is reported. CONCLUSION: In our patient, the ovary may have been seeded hematogenously by salmonellae and may have evolved into a local infection. PMID- 15108869 TI - Intrapartum antibiotic prophylaxis and early-onset neonatal sepsis patterns. AB - OBJECTIVE: To compare the relative effects of intrapartum antibiotic prophylaxis regimens on patterns of early-onset neonatal sepsis. METHODS: We performed an historical cohort study of 17187 infants born at our center from September 1993 to February 2000. A risk-based strategy was employed prior to July 1996 and a screening-based strategy was utilized thereafter. Ampicillin was utilized prior to March 1995 and penicillin was used thereafter. RESULTS: There were 75 cases of neonatal sepsis, 34 (4. 10/1000) in the risk-based era and 41 (4.63/1000) in the screening-based era (p = 0.62). There were fewer ampicillin-resistant isolates during the risk-based than the screening-based era (32 versus 61%; p = 0.014). The only significant change in organism-specific sepsis rates was an increase in the rate of infection caused by coagulase-negative staphylococci in the screening based era (0.36 versus 1.46/1000; p = 0.018), but 75% of infants infected with these organisms were not exposed to beta-lactam antibiotics within 72 h prior to delivery. For the risk- and screening-based eras, respectively, the rates of Gram negative sepsis (1.21 versus 1.46/1000; p = 0.65) and the proportions of Gram negative pathogens that were ampicillin-resistant (70 versus 77%; p = 1.0) were similar. The drug employed for prophylaxis did not appear to affect the pattern of sepsis cases. CONCLUSION: In our patient population, coagulase-negative staphylococci have become the most common cause of early-onset neonatal sepsis. The cause of this shift in pathogen prevalence is uncertain and seemingly unrelated to intrapartum antibiotic exposure. PMID- 15108870 TI - From birth to maturity: a group of patients who have completed their protocol management. Part I. Unilateral cleft lip and palate. AB - The optimal management of the cleft lip and palate patient from birth to completion of treatment presents a formidable challenge to the plastic surgeon and the associated health care system. The multidisciplinary team approach for the management of these patients is widely accepted. However, a paucity of literature exists discussing specific protocol management, interventions, and the long-term outcomes of patients who have completed a strict treatment protocol with a consistent multidisciplinary team. The aim of this study was to present the details of the specific management protocol at the Australian Craniofacial Unit for cleft lip and palate patients and to present a group of patients who have completed this specific protocol and discuss the details of their long-term care. During a 28-year period from 1974 to 2002, the records of 337 patients treated for unilateral cleft lip and palate were evaluated. Of these 337 patients, 22 have completed the same specific protocol management. The same surgeon (David, the senior author) has been responsible for performing all operative interventions and for overseeing the care of each of the 22 patients, ensuring that the treatment protocol has been executed appropriately and without deviation. The interventions and outcomes were analyzed on the basis of speech, hearing, nasal airway, occlusion, psychosocial adjustment, and appearance. Because of the large volume of data and potential differences in outcomes, the authors' intention is to present this as part I of a four-part series beginning with unilateral cleft lip and palate. The results of isolated cleft palate, isolated cleft lip, and bilateral cleft lip and palate will be presented as parts II, III, and IV, respectively. Speech results were assessed as normal speech, mild abnormality, or severe abnormality by objective measures, and intervention for velopharyngeal insufficiency was noted. Seventeen patients were rated as having normal speech. Four patients were rated as having mild speech abnormality, one patient was rated as having severe speech abnormality, and seven patients required surgery for velopharyngeal insufficiency. Hearing results were measured objectively, and good hearing results were obtained in 18 cases. Five patients required tympanoplasty. All patients required alveolar bone grafting. The high Le Fort I osteotomy was performed in six cases. Bimaxillary surgery was performed in one case. Of all the patients assessed from birth to maturity, 13 required between three and five surgical interventions, and nine required six operations or more. Further details and photographs of preoperative and postoperative examples are provided. PMID- 15108871 TI - Orthopedic intrusion of premaxilla with distraction devices before alveolar bone grafting in patients with bilateral cleft lip and palate. AB - Surgical repositioning of the downward displaced premaxilla in bilateral cleft lip and palate patients remains a controversial and perplexing issue because of its detrimental effects on the growth of the premaxilla. The purpose of this prospective clinical study was to introduce and evaluate the treatment results of an innovative technique for nonsurgically intruding the downward displaced premaxilla. Eight consecutive cases of bilateral cleft lip and palate at the age of mixed dentition were included for the correction of their premaxillary deformities. A pair of intraoral tooth-borne distraction devices was used for the orthopedic intrusion. Serial lateral and posteroanterior cephalometric radiographs were taken periodically for evaluating the growth of the premaxilla 1 year before the intrusion, changes during the intrusion, and growth/relapse up to 1 year after the intrusion. There was no overgrowth of the premaxilla or overeruption of the maxillary incisors during the 1-year observing period before the orthopedic intrusion. The treatment results revealed that the downward displaced premaxillae were all corrected within 1 month. Cephalometrically, 46 percent of the correction resulted from a true orthopedic intrusion and another 54 percent from a dentoalveolar effect in which the maxillary incisors were intruded and the premaxillary dentoalveolus was shortened. The cephalometric evaluations also implied that what occurred during the orthopedic intrusion was mostly the sutural contraction osteogenesis/osteolysis in the vomeropremaxillary suture combined with slightly mechanical upward displacement of the vomeronasal septum complex and nasal bones. The orthopedic intrusion of the premaxilla with distraction devices is an effective nonsurgical method for correcting the downward displaced premaxilla before alveolar bone grafting in patients with bilateral cleft lip and palate, and the results remained stable after 1 year. PMID- 15108872 TI - A new rig for standardized craniofacial photography put to the test. AB - This article describes and tests a photography rig that has been built at the University of Melbourne, Australia, specifically for the purpose of taking rapid and highly standardized craniofacial photographs, in simultaneous views of front and profile. The rig uses a novel projected light range-finding system that has been designed for easy and accurate positioning of subjects, in the natural head position, at precise distances from the frontal camera. Results of experiments examining the intraobserver error of multiple photographs taken on the rig indicate that high-quality, repeatable photographs can be taken after a reasonably large amount of time has lapsed between photography sessions (e.g., 30 days). This study also indicates that some variability remains between photographs even when highly standardized protocols are followed. Consequently, it is expected that the variation between photographs with limited standardization is much larger and more likely to cause significant errors in any comparisons. PMID- 15108873 TI - Infraorbital nerve injury associated with zygoma fractures: documentation with neurosensory testing. AB - Persistent sensibility abnormalities after correction of zygoma fractures indicate injury to the infraorbital nerve and may produce pain. To investigate this, a retrospective study of 25 patients who had undergone surgical correction of a zygoma fracture was performed. Bilateral neurosensory measurements were obtained with the Pressure-Specified Sensory Device (Sensory Management Services, Baltimore, Md.). Seven of the 25 patients had required orbital floor reconstruction. Each patient had undergone fracture correction at least 6 months earlier and was interviewed, at the time of sensibility testing, regarding symptoms related to the fracture. The data were evaluated by a blinded examiner, from a separate clinical facility, who attempted to predict the side of the fracture and the degree of zygoma displacement on the basis of measurements of sensibility of the paranasal, upper lip, and zygomaticotemporal areas. Seventy six percent of patients demonstrated abnormal sensibility on the side of the zygoma fracture, compared with the contralateral side. Sensibility was abnormal for 100 percent of the patients who required orbital floor reconstruction. Seventy-four percent of patients with abnormal sensibility reported symptoms related to the fracture. Eighty percent of the zygoma fractures were correctly identified, with respect to the side of the fracture, by the blinded examiner on the basis of the neurosensory measurements alone (p < 0.005). Predictions proved correct for 91 percent of the patients with widely displaced fractures and none of the patients with nondisplaced fractures. The results of this study suggest that neurosensory testing is an important clinical adjunct for the evaluation of patients with facial pain or dysesthesia after facial fracture reconstruction. The results suggest the need to develop algorithms for the diagnosis and treatment of trigeminal nerve injuries after craniofacial trauma. This approach could also be applicable to dysesthesia or pain after aesthetic facial surgical procedures. PMID- 15108874 TI - Nose reconstruction by chondrocutaneous preauricular free flaps: anatomical basis and clinical results. AB - Full-thickness defects of the nose result in considerable and distressing disfigurements. Ideally, reconstruction of such defects must be achieved in as few stages as possible and secondary, disfigurement kept to a minimum. In this study, the authors aimed to learn whether nose reconstruction could benefit from chondrocutaneous free flaps taken from the auricular tragus. In 72 ears, the vascular blood supply of the tragus was studied following injection of colored latex. Color-coded Duplex sonography served as a noninvasive method for demonstrating the blood supply of the target area. The procedure of nose reconstruction using the free chondrocutaneous tragus flap and the cosmetic results of this procedure in six patients are presented. Except for 2.8 percent of the anatomical specimens, the superficial temporal artery gave rise to the tragus and its overlying skin. The diameter of these branches ranged from 0.65 to 0.82 mm. Using the tragus composite free flap, the anatomical shape of the nose could be reconstructed successfully, and 6 months after surgery, the color and texture of the flap were very similar to those of the remaining nose. Using deeper parts of the tragus cartilage resulted in minimal scars and maintenance of the tragus anatomical shape. Free tragus flaps could be an alternative approach for nose reconstruction. PMID- 15108875 TI - What exactly was wrong with the Trilucent breast implants? A unifying hypothesis. AB - Trilucent soybean oil-filled breast implants were initially announced as the ultimate prostheses for breast augmentation. However, after an increasing number of reports of local complications and hazardous metabolites attributable to lipid oxidation, first the United Kingdom Medical Devices Agency and later the Belgian National Ministry of Health urged all plastic surgeons to contact their patients and advise them to have the implants removed and, if desired, replaced with another type of prosthesis. In our plastic surgery department, 13 patients received bilateral implants with triglyceride-filled prostheses between February and July of 1996, for primary breast augmentation or replacement of previously implanted prostheses. For 12 of those 13 patients, the prostheses have been explanted, because of unilateral breast enlargement attributable to a ruptured prosthesis for five patients and following the recommendation of the Belgian National Ministry of Health for the other seven patients. Before explantation, all patients underwent standard clinical examinations, with assessments of breast shape, volume, and firmness. Blood analyses were performed, with a special focus on liver enzymes, as were urinalyses. Magnetic resonance imaging scans were obtained before explantation; for two patients, the scans revealed a fluid level separating two liquid layers in an intact prosthesis. This is the first report of such a finding. The removed implants were examined for any damage or shell deterioration and for changes in color and viscosity, the weights and volumes were measured and compared with the initial values for the implanted prostheses, and complete biochemical analyses of the accumulated fluid in cases of ruptured prostheses and of the filler material in cases of intact prostheses were performed. This small but well-documented series illustrates the multitude of problems associated with triglyceride-filled implants, including bleeding of the triglyceride filler; shell deterioration, as indicated by a loss of texture and extreme fragility of the implant (with rupture or delamination with a simple finger touch); an increase in osmotic pressure exerted by the degraded filler material; progressive weakening of the outer silicone shell, with influx of plasma proteins of up to 750 kDa, eventually resulting in rupture of the prosthesis; a lack of oxidative stability and the formation of toxic oxidation products; a lack of biocompatibility, with the formation of insoluble organic soap-like material; and a pronounced inflammatory reaction. It is concluded that the sequential and/or simultaneous occurrence of (1) implant bleeding, (2) lipid infiltration of the silicone elastomer, and (3) inflammation attributable to oxidation products provides an overall explanation or unifying hypothesis for the wide variety of adverse events related to soybean oil-filled implants. PMID- 15108876 TI - Skin waste, vertex angle, and scar length in excisional biopsies: comparing five excision patterns--fusiform ellipse, fusiform circle, rhomboid, mosque, and S shaped. AB - The common excision skin pattern is either a fusiform ellipse or another pattern with dissimilar length and width. The purpose of this study was to define the most advantageous skin pattern regarding skin waste, vertex angle, and scar length. Five skin excision patterns used traditionally for closure of round lesions were analyzed: fusiform ellipse, fusiform circle, rhomboid, mosque, and S shaped. In the analysis, the pattern characteristics were formulated by geometric principles, from which the results were compared. The smallest skin waste was found in rhomboid and mosque patterns, whereas the largest skin waste was found in the fusiform circle and ellipse. The vertex angle was found to decrease monotonously with the excision length-to-width ratio for all patterns except the mosque shape, which is zero per definition. The paradigm stating that a vertex angle of 30 degrees or less is maintained for length-to-width ratios below 4 in the surgical ellipse was found incorrect. It holds only for rhomboid and S-shaped excisions. The scar length was found almost independent of the pattern, with a variance of 3 percent. The authors conclude that the most advantageous surgical skin patterns are the rhomboid and mosque excisions. PMID- 15108877 TI - Our experience with Wisebands: a new skin and soft-tissue stretch device. AB - Complex wounds that involve skin and soft-tissue defects that are unsuitable for primary closure by conventional suturing are common in the field of surgery. Among the many surgical options available to overcome these problems are various mechanical devices that have recently been proposed for delayed primary closure of such wounds. The authors present their experience with a new complex wound closure device, Wisebands, a device uniquely designed for skin and soft-tissue stretching. During the last 2 years, the authors have treated 20 patients with 22 skin and soft-tissue wounds for which primary closure was not feasible. The Wisebands devices were applied to the wounds, stretching the skin and underlying soft tissue, gradually closing the defects until the edges were sufficiently approximated for primary closure. Successful wound closure was achieved in 18 patients (90 percent). The Wisebands devices were removed in two patients (10 percent) because of major wound complications. In two other patients (10 percent), minor wound complications had occurred that did not necessitate removal of the device. At a mean follow-up of 1 year (range, 10 months to 2 years), stable scarring with no functional or significant aesthetic deficit was achieved. The authors conclude that the Wisebands device facilitates closure of complex skin and soft-tissue wounds, with low morbidity and complication rates, and can provide the surgeon with another important tool for closing complex wounds. Nevertheless, appropriate patient selection, intraoperative judgment, and close postoperative care are essential to ensure closure and avoid undue complications. PMID- 15108878 TI - Arterial and venous anatomical features of the pectoralis minor muscle flap pedicle. AB - The pectoralis minor muscle has been used as an innervated, vascularized, free muscle graft in the field of facial reanimation for 20 years. Throughout this period, several centers have demonstrated consistent success with functional muscle transfer; however, opinions regarding the arterial pedicle of the flap have varied. The lateral thoracic and thoracoacromial arteries have been proposed as the predominant arterial sources. It has been the experience of our unit that a vessel (not described in anatomy textbooks) arising directly from the axillary artery and entering the muscle from its dorsal surface provides the dominant supply to the flap and is capable of sustaining it for free-tissue transfer. The vascular pedicle encountered was recorded and photographed in 97 consecutive cases in which the pectoralis minor muscle flap was raised. The findings demonstrated that the dominant supply to the muscle was from a single artery in 77 percent of cases and took the form of an artery arising directly from the axillary vessel in 72 percent of cases. More than one major arterial source was noted in the remainder of the cases. The venous outflow was usually through single or multiple veins running directly from the muscle into the axillary vein. PMID- 15108879 TI - Irradiation after immediate tissue expander/implant breast reconstruction: outcomes, complications, aesthetic results, and satisfaction among 156 patients. AB - Chest wall irradiation is becoming increasingly common for mastectomy patients who have opted for immediate breast reconstruction with tissue expanders and implants. The optimal approach for such patients has not yet been defined. This study assesses the outcomes of a reconstruction protocol for patients who require irradiation after tissue expander/implant reconstruction. The charts of all patients who underwent immediate tissue expander/implant reconstruction at Memorial Sloan-Kettering Cancer Center between January of 1995 and June of 2001 and who had not previously undergone irradiation were retrospectively reviewed. A subgroup of patients who required chest wall irradiation after mastectomy and reconstruction was identified. Those patients were treated according to the following treatment algorithm: (1) reconstruction with tissue expander placement at the time of mastectomy , (2) tissue expansion during postoperative chemotherapy, (3) exchange of the tissue expander for a permanent implant approximately 4 weeks after the completion of chemotherapy, and (4) chest wall irradiation beginning 4 weeks after the exchange. All irradiated patients with at least 1 year of follow-up monitoring after the completion of radiotherapy were evaluated with respect to aesthetic outcomes, capsular contracture, and patient satisfaction. A control group of nonirradiated patients was randomly selected from the cohort of patients treated during the study period. During the 5-year study period, a total of 687 patients underwent immediate reconstruction with tissue expanders. Eighty-one patients underwent postoperative irradiation after placement of the final implant. A total of 68 patients who received postoperative chest wall irradiation underwent at least 1 year of follow-up monitoring after the completion of radiotherapy, with a mean follow-up period of 34 months. Seventy-five nonirradiated patients were evaluated as a control group. Overall, 68 percent of the irradiated patients developed capsular contracture, compared with 40 percent in the nonirradiated group (p = 0.025). Eighty percent of the irradiated patients demonstrated acceptable (good to excellent) aesthetic results, compared with 88 percent in the nonirradiated group (p = not significant). Sixty-seven percent of the irradiated patients were satisfied with their reconstructions, compared with 88 percent of the nonirradiated patients (p = 0.004). Seventy-two percent of the irradiated patients stated that they would choose the same form of reconstruction again, compared with 85 percent of the nonirradiated patients. The results of this study suggest that tissue expander/implant reconstruction is an acceptable surgical option even when followed by postoperative radiotherapy and should be considered in the reconstruction algorithm for all patients, particularly those who may not be candidates for autogenous reconstruction. PMID- 15108880 TI - Immediate toe-to-hand transfer in acute hand injuries: overall results, compared with results for elective cases. AB - In the past 5 years, 25 mutilated digits were reconstructed with immediate toe-to hand transfers after acute hand injuries, for 21 patients. The overall results of the immediate toe-to-hand transfers were evaluated and compared with the results of 65 elective procedures performed during the same period by the same surgeon. There were 15 cases of great toe-to-hand transfer for thumb reconstruction, two cases of second toe transfer for index finger reconstruction, and four cases of simultaneous two-toe transfer for reconstruction of multiple-digit amputations. Two cases (two of 25 cases, 8 percent) were successfully salvaged with emergency reexploration. The incidences of emergency reexploration and postoperative infection were not significantly different from those for elective toe-to-hand transfer cases. The duration of industrial insurance coverage was much shorter than for elective cases, averaging 225 days (p < 0.001). Approximately 44 percent of the patients maintained their original jobs after immediate toe-to-hand transfer. The subjective satisfaction self-assessment scores of aesthetic appearance and function for the newly reconstructed thumb averaged 80 and 88 (of a total score of 100), respectively. Although satisfaction was lower than for elective reconstruction (p < 0.001), it was higher than for reconstruction of other digits. The donor-site appearance after great toe harvesting was mostly unsatisfactory. Immediate toe-to-hand transfer provides many advantages over the elective procedure in acute hand injuries, including single-stage reconstruction, shortened convalescence, early return to work, and socioeconomic efficiency. Because there were no significant differences in the success rates, frequencies of complications, or ultimate functional results, immediate toe-to-hand transfer is a safe and reliable procedure that is indicated for specific cases of acute digital amputation. PMID- 15108881 TI - The surgical workforce crisis: rising to the challenge of caring for an aging America. AB - In the past century, both the field of surgery and the population it serves have markedly changed. The number of elderly individuals in the United States is rapidly increasing. However, this increase has not been accompanied by commensurate growth in the surgical workforce. As the demographic characteristics of medical students have become more diverse, medical student interest in surgical training has declined. These trends raise two fundamental questions. (1) Will there be enough surgeons to meet the needs of an aging United States population? (2) Who will these surgeons be? PMID- 15108882 TI - Alterations in apoptosis and epithelial-mesenchymal transformation in an in vitro cleft palate model. AB - The processes of apoptosis and epithelial-mesenchymal transformation have been identified as two major mechanisms by which secondary palatal shelves achieve fusion. The aim of this study was to investigate alterations in these mechanisms by changing the physical distance between paired palatal shelves in an in vitro model of palatogenesis. Wild-type palatal pairs were dissected from E13.5 CD1 mouse embryos and allowed to grow in tissue culture for 48 hours at various intershelf distances. During the fusion process, medial edge epithelial cell fate was assessed using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining, to evaluate apoptosis, and carboxyfluorescence (carboxy-2,7'-dichlorofluorescein diacetate succinimidyl ester) labeling, to measure transformation to mesenchymal cells. Palatal pairs separated in culture greater than or equal to 0.4 mm failed to fuse. TUNEL staining showed that the number of apoptotic cells in the palatal shelves increased as the intershelf distance increased, becoming marked in shelves that did not achieve fusion. The amount of epithelial-mesenchymal transformation, however, decreased with increasing intershelf distance. These results suggest that the contribution of epithelial-mesenchymal transformation and apoptosis to palatal shelf development and fusion can be altered by physical proximity. Therefore, one mechanism behind clefting in utero may result from an imbalance in epithelial-mesenchymal transformation and apoptosis as observed in vitro where palatal shelves are challenged to fuse by physical separation. This effect could be significant in the understanding and treatment of developmental palatal abnormalities. Perhaps in utero manipulation of intershelf spacing or epithelial-mesenchymal transformation and/or apoptosis could reverse the clefting paradigm. PMID- 15108883 TI - Epithelialization process of free fascial flaps used in reconstruction of oral cavity mucosa defects in dogs. AB - Bare free fascial flaps are increasingly used for restoration of soft-tissue defects of the oral cavity because they provide thin, foldable tissues with high epithelialization capacity to preserve local anatomy as well as chewing, phonation, and deglutition. However, there are unanswered questions regarding the epithelialization process and other histopathologic changes occurring after transfer of these flaps into the oral cavity. To investigate these changes thoroughly, an experimental study was conducted in the dog model. Bare dorsal thoracic fascia was used as the free flap model. Ten adult dogs were used in this experiment. Oral mucosa defects measuring 6 x 5 cm were created. Free dorsal thoracic fascia flaps were harvested. The vascular pedicle of the fascia flap was anastomosed with the superior thyroidal artery and external jugular vein. Then, the flaps were transferred into the mucosa defects. The dogs were divided into groups, each composed of two animals. At 7, 14, 21, 30, and 60 days postoperatively, general anesthesia was administered to the groups 1, 2, 3, 4, and 5, respectively. First, clinical assessment was performed; then specimens were obtained. Initially, the flaps were gradually infiltrated by acute inflammatory cells coming from the circulation and then replaced by granulation tissue. Epithelial cells deriving from wound margins migrated onto the granulating flaps with eventual coverage of highly organized epithelium after 4 weeks, and the fascia flap could not be differentiated from the native mucosa. The flaps were replaced by normally maturated fibrous tissue containing regular collagen fibers, instead of atypical scar tissue. Wound contraction was calculated as 18 percent at postoperative day 60. It was detected that bare free fascia flaps used in the repair of mucosa defects act as a scaffold and complete epithelialization from surrounding margins. They can be accepted as the main surgical option for the reconstruction of oral cavity mucosa defects. PMID- 15108884 TI - Role of nitric oxide in skin flap delay. AB - Surgical delay of skin flaps before transfer is known to improve flap viability. This study attempts to elucidate the mechanism of vasodilation by exploring the effects of nitric oxide on the microcirculation of delayed skin flaps. Using a skin flap model in 22 CD-1 white mice, the diameter of two nonterminal choke arteries was measured using in vivo videomicroscopy. Vessel flow was also measured using an optical Doppler velocimeter. Similar measurements were recorded in several animals on the same vessels in which subcutaneous dissection without elevation was performed. Average vessel diameter ranged from 21.77 to 25.55 microm before skin flap delay. Average flow ranged from 1.72 to 2.44 nl/sec before delay. Next, each animal received an intraperitoneal dose of nitro aminomethyl-1-arginine (L-NAME), a nitric oxide synthase inhibitor delivered by means of osmotic pump at a level of 0 (n = 13 arteries), 20 (n = 10), 50 (n = 8), or 100 mg/kg/day (n = 7). Flaps were re-elevated 72 hours later and the aforementioned measurements were repeated. Vessel diameter increased to 44.92 microm in the control (0 mg/kg L-NAME) animals. Flow increased to 7.66 nl/sec in the control animals. Vessel dilation and flow did not change significantly in the nonoperative vessels. As the dose of L-NAME increased in the treated animals, there was a significant decrease in vasodilation and flow (p = 0.015 and p = 0.03, respectively). The authors' results demonstrate that nitric oxide is an important element of vasodilation and contributor to the phenomenon of skin flap delay. PMID- 15108885 TI - Metabolic characteristics of experimental free vascularized canine gracilis muscle transfers. AB - A canine gracilis model was used to study muscle energy metabolism and enzyme activities after free vascularized muscle transfer. Fifteen male mongrel dogs underwent orthotopic, free transfer of the left gracilis with microneurovascular anastomosis. After a minimum of 10 months' recovery, muscle biopsy specimens were obtained from the transfers and the contralateral controls and analyzed for relative fiber type areas and maximum activities of phosphorylase, hexokinase, phosphofructokinase, glycerol-3-phosphate dehydrogenase (GPDH), pyruvate kinase, lactate dehydrogenase, citrate synthase, succinate dehydrogenase, 3-hydroxyacyl coenzyme A dehydrogenase (HAD), and creatine phosphokinase. Biopsy specimens obtained before and after a 10 minute, 20-Hz contraction were analyzed for glucose, glycogen, glycolytic intermediates, phosphocreatine, total creatine, and adenine nucleotides (adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, inosine monophosphate, and inosine). There was no significant transfer versus control difference in type I relative fiber area (45 +/- 4 percent versus 44 +/- 3 percent). Total creatine was significantly reduced in the transferred muscles relative to control (83.1 +/- 3.0 mmol/kg versus 100.6 +/- 5.1 mmol/kg dry weight). Maximal activities of phosphorylase, pyruvate kinase, lactate dehydrogenase, citrate synthase, succinate dehydrogenase, HAD, and creatine phosphokinase were diminished in transfers relative to controls, although hexokinase activity was significantly higher in the freely transferred gracilis muscles. During the 20-Hz contraction, muscle transfers produced less force initially, although the force/time integral over the 10-minute stimulation was similar in transfers (277 +/- 25 N/g/second) and controls (272 +/- 24 N/g/second). The contraction was associated with significant glvcogen use and lactate accumulation in both transfers and controls, although this was less pronounced for the transfers. Glycolytic flux appeared muted in the transfers relative to controls. Significant, similar high-energy phosphagen reductions and inosine monophosphate accumulation were noted during the contraction in both groups. Contractile activity is associated with the expected pattern of muscle metabolite changes following free vascularized transfer, indicating the components of cellular energy metabolism are not qualitatively altered after microneurovascular muscle transfer. In contrast, quantitative differences suggest that free vascularized muscle transfer can be associated with a muscle enzyme profile consistent with deconditioning and the presence of denervated muscles fibers in the absence of fiber type profile changes. PMID- 15108886 TI - Balloon expansion of the pectoralis major muscle flap in sternoplasty: a biomechanical and histologic study in a rat model. AB - The pectoralis major advancement flap is currently the most popular technique for reconstruction of the anterior chest in patients with sternotomy wounds. Recently, the SpaceMaker balloon was introduced for rapid expansion of the pectoralis major muscle intraoperatively. The aim of the present study was to investigate the biomechanical and histologic effects of this expansion technique in a rat model. The upper 2 cm of the sternum was resected in 54 male rats. Reconstruction with balloon-assisted pectoralis muscle expansion was performed in 24 rats (study group). Another 24 rats underwent reconstruction with simple muscle advancement without expansion. Submuscular insertion of a catheter for expansion, without inflation, was performed in the remaining six rats (sham group). Rats were killed either immediately or 2 to 4 weeks after surgery. Thirty eight rats, including 16 after reconstruction with expansion, 16 after reconstruction without expansion, and six in the sham group, were killed immediately after surgery. Sixteen rats were killed 2 to 4 weeks after surgery, eight rats for each reconstruction technique. Before the animals were killed, the biomechanical properties of the muscles were tested with weights to calculate stiffness (in newtons per meter) and compliance gain (in percent). After the animals were killed, biopsy specimens were obtained for histologic analysis. Results indicated significantly lower muscle stiffness in the study group compared with the others immediately after surgery (p = 0.0000), although the difference failed to achieve statistical significance 2 to 4 weeks later (p = 0.76). In the study group, the compliance gain was 74.4 percent immediately after surgery but only 3.4 percent 2 weeks to 1 month postoperatively. Histologic examinations in all groups immediately and 2 to 4 weeks after surgery revealed regular muscle striation with no signs of inflammation. The elastic stiffness of the rat pectoralis major muscle is significantly reduced following rapid intraoperative expansion and returns to normal 2 to 4 weeks later. PMID- 15108887 TI - PHEMA as a fibrous capsule-resistant breast prosthesis. AB - The presence of a silicone (poly-dimethyl siloxane) breast prosthesis in a breast reconstruction patient typically leads to fibrous tissue encapsulation of the prosthesis. Fibrous capsular contracture forces the prosthesis into a hardened sphere. The initially satisfactory cosmetic result can thus be changed into a deformed mass of inappropriate compliance. It is the author's hope with the present study to identify a material for implantation with a diminished tendency to form fibrous encapsulation, to improve the long-term results of prosthetic implants. The purpose of this investigation was to compare the early capsule production quality of poly-2-hydroxyethyl methacrylate (PHEMA) and poly-dimethyl siloxane (silicone). Each of five rats subcutaneously underwent implantation with both a disk of poly-dimethyl siloxane (control) and a similar disk of PHEMA. In this study, the extent of fibrous encapsulation was assessed at 6 weeks after implantation of the two disk types. The five disks of poly-dimethyl siloxane were embedded in fibrous tissue, whereas there was no apparent fibrous tissue surrounding the implants of PHEMA. The author concludes that the results for PHEMA were superior to those for silicone at 6 weeks with regard to fibrous encapsulation (p = 0.0312). PMID- 15108888 TI - Validation of a model for the study of multiple wounds in the diabetic mouse (db/db). AB - The genetically diabetic db/db mouse exhibits symptoms that resemble human type 2 diabetes mellitus, demonstrates delayed wound healing, and has been used extensively as a model to study the role of therapeutic topical reagents in wound healing. The purpose of the authors' study was to validate an excisional wound model using a 6-mm biopsy punch to create four full-thickness dorsal wounds on a single db/db mouse. Factors considered in developing the db/db wound model include reproducibility of size and shape of wounds, the effect of semiocclusive dressings, comparison with littermate controls (db/-), clinical versus histologic evidence of wound closure, and cross-contamination of wounds with topically applied reagents. The size of wounds was larger, with less variation in the db/db mice (31.11 +/- 3.76 mm2) versus db/- mice (23.64 +/- 4.78 mm2). Wounds on db/db mice that were covered with a semiocclusive dressing healed significantly more slowly (mean, 27.75 days) than wounds not covered with the dressing (mean, 13 days; p < 0.001), suggesting the dressings may splint the wounds open. As expected, wounds healed more slowly on db/db mice than db/- mice (covered wounds, 27.75 days versus 11.86 days, p < 0.001; wounds not covered, 13 days versus 11.75 days, p = 0.39). Covered wounds, thought to be closed by clinical examination, were confirmed closed by histology only 62 percent of the time in the db/db and 100 percent of the time in the db/- mice. Topical application of blue histologic dye or soluble biotinylated laminin 5 to one of the four wounds did not spread locally and contaminate adjacent wounds. Multiple, uniform, 6-mm wounds in db/db mice heal in a relatively short time, decrease the number of animals needed for each study, and allow each animal to serve as its own control. The db/db diabetic mouse appears to be an excellent model of delayed wound healing, particularly for studying factors related to epithelial migration. PMID- 15108889 TI - Bipolar anastomosis technique with removable instruments: an easy, fast, and reliable technique for vascular anastomosis. AB - The interrupted suture technique is most commonly used for microsurgical vascular anastomosis. For several reasons (e.g., exposure of suture material to blood, time needed), many attempts have been made to find other solutions. This article describes a new means of performing a microsurgical vascular anastomosis. The aim of this study was to show the feasibility and possible advantages of this new technique. The basic components at work here are a modified cuff and electrically generated heat used to unite the vessel walls. In this way, both endothelial layers are adapted without manipulating the inside of the vessel or leaving behind foreign matter. Various energy/coagulation time settings were used to perform arterial anastomoses (n = 42) in an isogeneic abdominal aorta interposition model in the rat. The quality of anastomosis was evaluated at days 1, 10, 21, and 120. Immediately after the welding process all anastomoses (n = 42) were patent. No stenosis was found at any observation time. Anastomosis time ranged from 3 to 18 minutes (average, 11 minutes). This new technique permits a vascular anastomosis to be performed easily and reliably with a high patency rate. With this technique, the authors are convinced that a skilled surgeon can create a high-quality anastomosis in a fraction of the time needed to sew an anastomosis. PMID- 15108890 TI - Bilateral cauliflower ear deformity: an unusual presentation of cutaneous Rosai Dorfman disease. AB - This case illustrates the variety of clinical presentations of Rosai-Dorfman disease. In this case, the disease presented as bilateral cauliflower ear deformity and was diagnosed on the basis of typical pathological findings. Although the cause of this disease is not fully understood, it is hoped that, with increased awareness and the identification of more cases, more questions may be answered with respect to this interesting condition that was so elegantly described by Rosai and Dorfman more than three decades ago. PMID- 15108891 TI - Juvenile active ossifying fibroma with massive involvement of the mandible. AB - Fibro-osseous lesions of the maxillofacial complex are often difficult to diagnose from both a clinical and a histopathologic point of view. The parameters for the diagnosis of juvenile active ossifying fibroma are as follows: a patient under 15 years of age, localization of the tumor, the radiologic aspect, and the tendency to recur. Although many authors favor conservative surgery rather than radical en bloc resection, immediate recurrence characterized by a high aggressive growth rate and the absence of a distinct separation between the tumor and the adjacent bone requires ex- tensive surgery, with wide demolition of the involved bone. PMID- 15108892 TI - Mammary hypoplasia resulting from hormone receptor deficiency. PMID- 15108893 TI - Reepithelialization of a full-thickness burn from stem cells of hair follicles micrografted into a tissue-engineered dermal template (Integra). AB - We report a head and neck full-thickness burn injury that was reconstructed with a tissue-engineered dermal template and then early implantation of microdissected hair follicles through the silicone epidermis 12 days after the burn injury. The treatment resulted in complete reepithelialization and a hair-bearing scalp without the need for a split-thickness skin graft. Restoration of the stem cell population, hair growth, and earlier reepithelialization were achieved using this novel micrografting technique, and histologic examination confirmed maturation of a normal skin type over the subsequent 2 years. PMID- 15108894 TI - Endotracheal tube fixation to the maxilla in patients with facial burns. PMID- 15108895 TI - A new concept and technique for reconstructing skin defects in the cheek region: an unfolded cube advancement flap. PMID- 15108896 TI - Periareolar approach for the correction of congenital symmastia. AB - This report elucidates the continued and relatively rare problem of congenital symmastia and its surgical repair without concomitant bilateral breast reduction. This case highlights the use of suction-assisted lipectomy techniques to address the excess fat in the presternal web and a periareolar approach for access to the intermammary space. The periareolar incision allows for the use of a concealed approach and the avoidance of a central scar that could result in hypertrophy or keloid formation, especially in this patient who is more prone to hypertrophic scarring. Furthermore, plication of the central web dermis to the sternal periosteum in a more superior position serves to not only correct the symmastia but also redrape the excess skin and restore the blunted inframammary folds. PMID- 15108897 TI - The pubic area skin and dermal graft donor site. PMID- 15108898 TI - Mast cells and hemangioma. AB - Hemangioma is a primary tumor of the microvasculature in which angiogenesis is initially excessive, followed by spontaneous regression of the newly formed vessels, with the cellular parenchyma gradually being replaced with fibrofatty tissue. Mast cells, which are highly heterogenous in terms of their morphology, function, and metabolic products, have been implicated in the pathophysiology of hemangioma. Csaba stain shows that mast cells are predominantly of the biogenic amine phenotype throughout the development of hemangioma. The predominance of this phenotype remains unaltered following successful steroid therapy, although their number increases fourfold. Mast cells, all of which stain positive for tryptase, and those that stain positive for chymase as well, have been identified in hemangioma biopsy specimens throughout the three developmental phases. The total number of mast cells is highest during the involuting phase, less in the involuted phase, and least in the proliferative phase. The proportion of mast cells that contain both tryptase and chymase decreases from the proliferative through involuting to the involuted phase. This decreasing proportion of mast cells that contain both tryptase and chymase with ongoing involution parallels that of progressive deposition of the extracellular matrix as indicated by increasing fibrosis and fatty deposition. The short-chain type VIII collagen, thought to play a key role in angiogenesis, has been detected throughout the developmental phases of hemangioma. It has been postulated that this collagen, which is produced early in new vessel development, provides a substratum to facilitate the migration of endothelial cells. It may also facilitate the deposition of other extracellular constituents and influence cell movement and the maintenance of cell phenotypes. The intracellular localization of type VIII collagen in mast cells only in the early proliferative phase suggests that there is an active synthesis by mast cells during this phase. The increasing extracellular localization during hemangioma development may be caused by an increased secretion of protein from intracellular stores. The increased number of mast cells during the involuting phase indicates that these cells may play a role in the regression of hemangioma. This is in contrast to the large body of evidence showing the proangiogenic role of mast cells. The proportion of proliferating mast cells decreases, whereas the proportion of mast cells positive for clusterin/apolipoprotein J increases with ongoing involution of hemangioma. Clusterin/apolipoprotein J expression has been considered as a prominent marker of apoptotic cell loss. The presence of clusterin/apolipoprotein J granules both in the adjacent endothelial cells and in capillary lumens suggests that mast cells may be secreting this apoptotic modulator to promote the regression of hemangioma. Certain effectors produced by mast cells may participate in the development of hemangioma. It has been proposed that one of the functions of mast cells is to release factors leading to the regression of hemangioma. The evidence suggests that although mast cells may have a function in the endothelial proliferation in hemangioma, they also play a crucial role in the regression of this tumor. However, the roles of mast cells in the life cycle of hemangioma are likely to be complex and may involve stimulators of angiogenesis in the proliferative phase but inhibitors in later phases. PMID- 15108899 TI - Boomerangs and international plastic surgery. PMID- 15108900 TI - Development of a novel nonantigenic dermal implant composed of human placental collagen. AB - Injectable bovine collagen has proven to be safe and effective for the treatment of contour defects for more than 20 years. After intradermal exposure to bovine collagen, the most commonly reported side effect is hypersensitivity (incidence of approximately 3 percent to test and approximately 1 to 2 percent to subsequent treatment). The main purpose of this study was to evaluate tissue response and antibody production in bovine collagen-sensitive patients who were treated with human collagen (predominantly type I) implant. Twenty-seven patients with confirmed hypersensitivity to bovine collagen received a depot of human collagen implant and then were treated for facial contour defects on two to five separate occasions over a 9- to 12-month period and followed through 36 months. Measurement of antibody titers indicated that none of the subjects receiving human collagen implant developed antibodies against human collagen, even in the presence of positive antibody titers against bovine collagen. Histologic examination of the depot sites in these patients showed only mild inflammation. These findings indicate that treatment with human collagen did not elicit an allergic response in these subjects who had confirmed hypersensitivity to bovine collagen. PMID- 15108901 TI - Trilucent breast implants: voluntary removal following the Medical Device Agency recommendation. Report on 115 consecutive patients. AB - In June of 2000, the U.K. Medical Device Agency recommended the removal of Trilucent implants as a precautionary maneuver in response to reports of local inflammatory reactions. This decision allowed the authors to operate on 115 consecutive patients between June of 2000 and January of 2001. On the preoperative examination, the authors found a very high incidence of rippling (66 percent), whereas capsular contracture was seen in only three patients (2.6 percent). Rippling was significantly more common in patients with subglandular implants. Five implants were found ruptured during the operation. This figure, together with the relative ease of implant breakage at removal, shows a premature deterioration of the implant shell. The authors also comment on implant bleeding, which seems common in this type of breast implant. The authors think that this is a possible cause for the rippling phenomenon, resulting from a reduction of the implant content. On the basis of these findings, the authors conclude that Trilucent implants are associated with a poor cosmetic outcome and a high rate of complications. PMID- 15108902 TI - Extended upper blepharoplasty for lateral hooding of the upper eyelid using a scalpel-shaped excision: a 13-year experience. AB - Excess skin of the upper lids is often accompanied by lateral overlap of skin with crow's feet because of the absence of fixation to the tarsal plate, giving the eye a sad, heavy look that often disturbs the lateral visual field. The accepted crescent-shaped blepharoplasty is somewhat convex, which is widest at the center of the lid with or without a lateral extension. However, in patients who have normal brow position or minimal eyebrow ptosis and whose main concern is the excess upper eyelid skin and lateral hooding, such a crescent excision may not suffice. A scalpel-shaped excision that is widest laterally and that tapers to a point medially will extirpate the maximal skin where it is most needed and overcome the skin excess in the lateral aspect of the upper lid. Between 1990 and 2002, 301 white patients (275 women and 26 men) between the ages of 33 and 79 years were operated on using the extended scalpel-shaped upper blepharoplasty technique. The follow-up period was more than 1 year. The lower margin of the incision is along the supratarsal crease, about 10 mm above the ciliary line. It begins medially about 1 cm above and lateral to the medial canthus. Above the lateral canthus, the skin marking is gently curved upward and outward, often within a natural skin crease or crow's feet to reach a little below and slightly beyond the lateral extremity of the eyebrow. The upper border of the incision joins the two extremities of the skin outline in a gentle convex curve. The general outline of the incision takes on the shape of a number 20 scalpel blade in which the maximal width is located laterally. Following excision of the excess skin and removal of protuberant fat pads if needed, suturing is executed from lateral to medial. The final suture line is in the form of an oblique flattened lazy S. Following the removal of the stitches on the fifth postoperative day, no wound dehiscence was noticed at the lateral scar zone. In the older individuals, due to the lax skin, the scar becomes scarcely noticeable with time and often falls within a pre-existent crow's feet crease. Elimination of some of the crow's feet was also demonstrated. In patients with visual field impairment, significant functional and visual improvement was achieved. Most patients mentioned a pleasing postoperative open "Oriental" look of the eyes. The extended scalpel shaped upper blepharoplasty adequately deals with the hooding of the skin laterally. This technique overcomes the excess of skin in both vertical and horizontal directions, since in suturing the lateral part of the skin defect in an oblique plane, slack skin is taken up transversely, and the technique provides some indirect upward support to the lateral eyebrow. In the absence of crow's feet in the younger person, this technique is not recommended because the lateral part of this suture line is visible, especially if the scar widens. PMID- 15108903 TI - The subdomal graft. PMID- 15108904 TI - A technique of brachioplasty. AB - Various techniques for the management of upper extremity contour deformities have been suggested since aesthetic brachioplasty was first described. Such deformities are commonplace with aging, after normal weight loss, and especially after massive weight loss such as is seen following bariatric surgery. Despite the multiplicity of procedures described for the correction of these deformities, there are still problems associated with current brachioplasty techniques, including incorrectly placed incisions, widened hypertrophic scars, and postoperative contour deformities. In addition, postoperative skin laxity and ptosis in the axillary region are frequently encountered in the more extreme deformities. The authors present their technique for upper extremity brachioplasty. This technique is suitable for patients with severe brachial ptosis and skin laxity, with relatively little lipomatous tissue, which may extend from the olecranon to the chest wall. The described surgical approach provides excellent overall extremity contour with favorable scars while simultaneously addressing axillary contour deformities. PMID- 15108905 TI - Irritable bowel syndrome and abdominoplasty. AB - Irritable bowel syndrome is a chronic disease involving pain or discomfort relieved by defecation and associated with a change in frequency or consistency of the stools. The effect of abdominoplasty on patients with irritable bowel syndrome has not been elucidated, so advising patients with irritable bowel syndrome about the effects of surgery on their disease was difficult. One hundred female patients from a pool of 120 patients responded to a questionnaire relating to abdominoplasty surgery. Follow-up ranged from 6 months to 2 years. Patients completed questionnaires formulated on the basis of Rome II Diagnostic Criteria. Of the 100 patients, nine had true irritable bowel syndrome, nine had moderate symptoms and were receiving medication (not true irritable bowel syndrome), 16 had mild symptoms on occasional medication, and 66 had no symptoms of irritable bowel syndrome before surgery. Of the true irritable bowel syndrome patients, all had symptomatic improvement with decreased medication, eight of the nine patients with moderate symptoms improved markedly, and five of the 16 patients with mild symptoms improved significantly. No patient had any initiation or deterioration of symptoms. It is thus concluded that the symptoms of irritable bowel syndrome are certainly not worsened, in the short term, by surgery, and may be alleviated or improved in most significant cases. PMID- 15108906 TI - Aesthetic outcome of breast implant removal in 85 consecutive patients. PMID- 15108907 TI - The see-and-treat clinic in plastic surgery: an efficient, cost-effective, and training-friendly setup. PMID- 15108908 TI - Invasive fungal sinusitis. PMID- 15108909 TI - Endoscopic brow lifts: making bone tunnels safer for sutures. PMID- 15108910 TI - Acquired strabismus following cosmetic blepharoplasty. PMID- 15108911 TI - Interrupted "waved round block suture" to secure skin grafts on the scalp. PMID- 15108912 TI - Dermatophytic pseudomycetoma of the scalp. PMID- 15108913 TI - Correction of full-thickness defects of the auricular scapha following Mohs surgery. PMID- 15108914 TI - Earlobe morphology delineated by two components: the attached cephalic segment and the free caudal segment. PMID- 15108915 TI - Nasal herpes after rhinoplasty. PMID- 15108917 TI - Augmentation rhinoplasty with dermal graft. PMID- 15108916 TI - Cleft lip/nose deformity and rhinolith. PMID- 15108918 TI - Late presentation of alloplastic implant extrusion. PMID- 15108919 TI - Surgical treatment of a seldom-seen craniofacial deformity. PMID- 15108920 TI - Axillary nerve injury in axillary block. PMID- 15108921 TI - An unusual lipoma. PMID- 15108922 TI - Lichen planus cutis and squamous cell carcinoma. PMID- 15108923 TI - Gestational gigantomastia after reduction mammaplasty. PMID- 15108924 TI - Concomitant open cholecystectomy and mammary reconstruction with bipedicled TRAM flap. PMID- 15108925 TI - Subfascial breast implant. PMID- 15108926 TI - Is the umbilicus truly midline? PMID- 15108927 TI - Circumcision in unqualified hands: a significant risk of complication. PMID- 15108928 TI - Osteocutaneous forearm flaps: the advantages of a preformed lightweight cast. PMID- 15108929 TI - Protection of dressings and wounds by cling film. PMID- 15108930 TI - The safety of rolled Penrose digital tourniquets for use in local procedures. PMID- 15108931 TI - The Hull hand table: a cost-effective alternative. PMID- 15108932 TI - The digital arteries proper arising from the dorsal metacarpal artery with a minor contribution from a persistent median artery: anatomical variation. PMID- 15108933 TI - A see-through in vitro tendon repair model. PMID- 15108934 TI - A simple salvage technique for single-stage, soft-tissue coverage of plantar first metatarsal head ulcerations and ablation of great toe osteomyelitis. PMID- 15108935 TI - Massive expanding lipoma of the toe. PMID- 15108936 TI - Plantar verrucous carcinoma continues to be mistaken for verruca vulgaris. PMID- 15108937 TI - Reconstruction aplasia cutis congenita (group V) of the trunk in a newborn. PMID- 15108938 TI - Bio-Alcamid: an electron microscopic study after skin implantation. PMID- 15108939 TI - The application of Steri-Strips. PMID- 15108940 TI - Use of fine forceps as a surgical marker. PMID- 15108941 TI - Technique for skin graft sizes. PMID- 15108942 TI - How many rats? PMID- 15108943 TI - As many as are needed. PMID- 15108944 TI - Bibliography. Current world literature. Hormones, autacoids, neurotransmitters and growth factors. PMID- 15108945 TI - Bibliography. Current world literature. Circulation and hemodynamics. PMID- 15108946 TI - Communication needs, methods, and perceived voice quality following head and neck surgery: a literature review. AB - Patients with head and neck cancer experience complex and frustrating communication problems after surgery, yet patient communication during the in hospital postoperative period has received relatively little attention in clinical and research literature. A computerized and hand search of the medical (MEDLINE, Cancerlit), psychological (health and psychosocial instruments), and nursing (CINAHL) literature (1968 to April 2002) produced 10 published studies and 1 clinical case report specifically addressing the communication needs, methods, or perceived voice quality of patients with head and neck cancer during the postoperative period (< or = 12 months after surgery). This review presents a summary and critique of research and related literature on in-hospital postoperative communication with adult patients who have head and neck cancer. Three major themes are addressed: (1) information needs, (2) communication methods and perceived voice quality and (3) quality-of-life perceptions related to communication, disfigurement, and socialization. This review shows that the communication needs, communication methods, and perception of voice quality among patients with head and neck cancer have been ignored during the in-hospital period. Clinical issues and technological advancements in augmentative and alternative communication applicable to the in-hospital period are discussed, and research implications are presented. PMID- 15108947 TI - Determinants of functional recovery in older adults surgically treated for cancer. AB - The purpose of this study was to examine the psychosocial and disease-specific factors that influence functional recovery in older adults newly diagnosed with cancer. Multivariate logistic regression models were estimated using panel data from a sample of community-residing adults older than 65 years surgically treated for lung, prostate, breast, or colorectal cancer (N = 172). Data were obtained between 1993 and 1997 during interviews 4 to 6 weeks after cancer surgery for Wave 1 and 14 to 16 weeks after hospital discharge for Wave 2. The outcome measure, functional recovery, was determined by comparing the physical function and physical role subscales of Medical Outcomes Study (MOS) SF-36 over time. Findings showed that prostatectomy patients were more likely to recover by Wave 2 when compared to individuals with lung, colon, or breast resections. Comorbidities and symptom severity were each significantly associated with a decreased probability of recovery. Pain and fatigue were the most common and most severe symptoms reported, regardless of primary site. Psychological well-being was a significant factor influencing functional recovery when age, comorbidities, site of disease, and symptom severity were controlled. The results clearly point to the need for psychological support following cancer surgery. PMID- 15108948 TI - The effect of childhood cancer on Hong Kong Chinese families at different stages of the disease. AB - In Hong Kong the incidence of cancer is 144.3 per 1,000,000 children and is the first cause of disease-related death among children aged between 1 and 14 years. The diagnosis of cancer in a child poses a threat to the family unit, with many families experiencing difficulties as they cope with the demands of their child's illness. The purpose of this descriptive study was to explore the effects of cancer on the child, parents, and siblings during different stages of the disease. An in-depth individual interview was undertaken with 13 Chinese parents, 10 children, and 11 well siblings. Thematic analysis of the qualitative data identified 5 key areas including knowledge and understanding of the nature of the disease, being truthful, responses to the disease, changing family relationships, and changes in family life. This article focuses on the findings of 2 of these key areas: knowledge and understanding of the nature of the disease and being truthful. The findings indicate that all parents were fully aware of the diagnosis, treatment, and prognosis of their child's disease. The child's knowledge of the disease varied, depending in part on parents' willingness to tell their children about the disease and its prognosis. Most parents were reluctant to share information about the disease with their children. Children, however, were found to have considerably more knowledge about the disease than parents reported, indicating a gap in communication between parents and children. The findings indicate the importance of disclosure about the disease within the family and the need for practitioners to facilitate information sharing among family members within this population group. PMID- 15108949 TI - Postoperative arm massage: a support for women with lymph node dissection. AB - PURPOSE/OBJECTIVE: To evaluate the usefulness of arm massage from a significant other following lymph node dissection surgery. DESIGN: Randomized clinical trial with a pretest-posttest design. Data were collected prior to surgery, within 24 hours post surgery, within 10 to 14 days post surgery, and 4 months post surgery. SAMPLE: 59 women, aged 21 to 78 undergoing lymph node dissection surgery and who had a significant other with them during the postoperative period. METHODS: Subjects were randomly assigned to intervention and control groups. Subjects' significant others in the intervention group were first taught, then performed arm massage as a postoperative support measure. RESEARCH MAIN VARIABLES: Variables included postoperative pain, family strengths and stressors, range of motion, and health related costs. FINDINGS: Participants reported a reduction in pain in the immediate postoperative period and better shoulder function. CONCLUSION: Arm massage decreased pain and discomfort related to surgery, and promoted a sense of closeness and support amongst subjects and their significant other. IMPLICATION FOR NURSING PRACTICE: Postoperative massage therapy for women with lymph node dissection provided therapeutic benefits for patients and their significant other. Nurses can offer effective alternative interventions along with standard procedures in promoting optimal health. PMID- 15108950 TI - Issues in Internet survey research among cancer patients. AB - Considering the increasing number of cancer patients who are online, it is clear that the Internet will provide an important research medium and/or setting for oncology nurses in the near future. Despite increasing Internet usage in nursing research and practice, issues in using the Internet among cancer patients as a research tool have rarely been explored and discussed. The purpose of the article is to propose future directions for Internet research among cancer patients based on discussions of practical issues raised in an Internet survey study among 40 online cancer patients. The issues raised through the research process include (a) ethical issues, (b) recruitment issues, (c) issues in Web site development and maintenance, and (d) data entry and analysis issues. On the basis of the discussions of these issues, some future directions for Internet survey studies are proposed, including dealing with ethical issues, getting computer expertise, using motivational strategies, and using national and international approaches. PMID- 15108951 TI - The impact of managed health care in the United States on women with breast cancer and the providers who treat them. AB - This article describes how the increasing shift to managed care has impacted the treatment of women with breast cancer, from the perspectives of patients and providers. A descriptive exploratory pilot study was undertaken in a comprehensive cancer center in an urban northeastern city of the United States. The use of General Systems Theory is used to describe how the health care system has adapted to recent insurance changes. Qualitative interviews with patients,physicians, nurses, and business office staff reveal that the type of insurance did not affect treatment decisions. Findings from this pilot study, however, reveal that the increasing shift to managed care has resulted in a change in roles for nurses, fragmentation of care, and intangible costs to the patients. PMID- 15108952 TI - Symptom occurrence, symptom intensity, and symptom distress in patients undergoing high-dose chemotherapy with stem-cell transplantation. AB - The main aim of this study was to investigate the patients' self-reported symptom occurrence, symptom intensity (SI), and symptom distress (SD) from admission for stem-cell transplantation (SCT) until discharge from the ward. Forty-three patients participated and data were collected at 7 different time-points by using the self-administered Symptom Frequency, Intensity, and Distress questionnaire for SCT (SFID-SCT). The results showed that symptom occurrence followed a curve on which the highest frequencies of symptoms were reported from the day of the SCT (T2) until the end of the protective care period (T5). The mean SI and SD scores became higher when the number of reported symptoms increased. Between T2 and T5, 33% to 54% of the patients reported >10 simultaneous symptoms. Symptoms reported by more than 50% of the patients during T2-T5 were tiredness, loss of appetite, mouth dryness, nausea, sleeping disturbances, diarrhea, and changes of taste. Loss of appetite, tiredness, and mouth dryness were, in descending order, the 3 symptoms reported as most intense and distressing. A statistically significantly higher SD-score was found for the patients undergoing allogeneic SCT on the day before start of the conditioning regimen, as compared to the patients undergoing autologous SCT. Patients reporting no anxiety on admission were found to have higher, mean SD-scores at the end of the hospital stay than anxious patients. The SFID-SCT questionnaire was found to give useful information not only about symptom occurrence but also about SI and SD. To use an instrument that distinguishes between these aspects of the symptom experience may help health care professionals to support the patients through the SCT-process. PMID- 15108953 TI - Patients' experiences of palliative care in the home: a phenomenological study of a Swedish sample. AB - In recent times the rapid expansion of interest in palliative care has become a significant feature of health care development. Caring in the palliative way means simultaneous attention to the medical, nursing, spiritual, emotional, and social needs of the patient. The main purpose of the present study has been to obtain an understanding of patients' experiences of palliative care at home with service from district nurses. The research design is influenced by Giorgi's phenomenology. Six patients diagnosed with cancer and receiving palliative care at home were interviewed and the transcribed interviews were analyzed. Interviews were conducted in the patients' homes. The findings show that the essential meaning of the patients' experiences of palliative home care can be described as "uncertain safety." This meaning of essence is explicated by 4 themes, which are labeled "Safe but unsafe at home," "A sense of powerlessness," "Change of everyday life," and "Hope and belief in the future." The findings of the study point out the importance of well functioning teamwork and resources to facilitate patient's experiences of safety in their own homes. It should be noted that the patient and their next of kin are members of the team. PMID- 15108954 TI - The relationship between factors that impair wound healing and the severity of acute radiation skin and mucosal toxicities in head and neck cancer. AB - PURPOSE: To determine which wound-healing factors impact on the severity of radiation skin and oral mucosal reactions in head and neck cancer and to test modifications to the Radiation Therapy Oncology Group (RTOG) acute toxicity scoring system. METHODS: A consecutive sample of 53 head and neck cancer patients who were scheduled for curative or palliative radiation therapy. Therapy was planned using traditional computerized techniques. A new RTOG subscale for tongue reactions was developed. Information on potential predictors was collected during the first week of treatment. Reactions were observed and documented each week throughout treatment using the RTOG Acute Reaction Scoring System scores of acute oropharyngeal reactions and various personal factors. RESULTS: Significant relationships were found between severe skin and oral reactions and age, commencing radiation within 2 months of surgery and smoking. Significant relationships for severe oral mucosal reactions were found with weight at the commencement of treatment, inadequate or poor diet, having had mucositis with previous chemotherapy, and the use of a custom-made Perspex tongue immobilizer. CONCLUSIONS: Three conclusions can be derived from this study: (1) structures within the oral cavity should be considered separately for toxicity scoring, (2) the newly developed tongue RTOG subscale adds accuracy and specificity to the RTOG acute toxicity scoring system, and (3) wound healing factors are an important component of understanding risk for side effects in head and neck cancer treatment. PMID- 15108955 TI - Evaluating key dimensions of the breast care nurse role in Australia. AB - A survey of women who had surgery for breast cancer during 1997-1998 at The St George Public and St George Private Hospitals identified substantial deficits in information provision, perceived support, and use of available services. Consequently, a breast care nurse (BCN) was appointed for a 12-month trial to assess women's needs and provide emotional support, education, and referral to appropriate services from diagnosis through to completion of treatment, according to evidence-based guidelines set out by the Australian National Breast Cancer Centre. A questionnaire regarding the role of the BCN was sent to 63 women who had surgery at either hospital from September 1999 to January 2000. Fifty women responded, for a 79% response rate. Ninety percent of respondents reported that it was helpful to talk to the BCN and rated verbal information given as helpful. Women accessed significantly more services than reported in 1998, and disparities in service use by age and public/private sector were eliminated. These results show strong consumer endorsement of service provision by the BCN. During this 12 month trial, consumer demand resulted in service expansion from 24 hours per week to 80. Ongoing evaluation of this developing role is required to ensure that women's needs are managed optimally. PMID- 15108956 TI - Spouses' perceptions of and reactions to living with a partner who has undergone surgery for rectal cancer resulting in a stoma. AB - The aim of this study was to illuminate spouses' perceptions of living with a partner who had been diagnosed with rectal cancer followed by surgery resulting in a stoma. The focus group interview was chosen as method of data collection. Two focus group interviews consisting of 2 meetings each were conducted. A qualitative content analysis was used. Five themes emerged: difficulties of being involved; living with uncertainty; learning to live in a new way; the altered body; and the search for explanations. There were feelings of anxiety about the partner's serious condition and the possible spread of the cancer. After the rehabilitation period, the stoma influenced the spouses' lives in many ways but they coped with the problems together with their partners and helped them adapt to their changed circumstances. This study indicates that the spouses were very much involved in caring for the patient suffering from rectal cancer by protecting them and showing empathy. However, the dependence on the stoma affected and restricted their family and social life. PMID- 15108957 TI - Anesthetic effects on glutamatergic neurotransmission: lessons learned from a large synapse. PMID- 15108958 TI - One thing leads to another. PMID- 15108959 TI - Trigger-dependent gene expression profiles in cardiac preconditioning: evidence for distinct genetic programs in ischemic and anesthetic preconditioning. AB - BACKGROUND: DNA chips facilitate genomic-wide exploration of gene expression. The authors hypothesized that ischemic (IPC) and anesthetic preconditioning (APC) would differentially modulate gene expression in hearts. METHODS: Affymetrix rat U34A gene chips were used to explore the transcriptional response to IPC and APC, sustained ischemia (110 min) without reperfusion, and time-matched perfusion in isolated rat hearts. IPC was induced by three cycles of 5 min of ischemia, and APC was induced by 1.5 minimum alveolar concentration isoflurane (110 min). For each heart, a separate chip was used for hybridization. Data were analyzed for significant > or = 2.0-fold changes in gene expression. Microarray results were confirmed by quantitative real-time reverse-transcription polymerase chain reaction. RESULTS: Of the 8,799 genes represented on U34A, 217 transcripts in the APC group, 234 in the IPC group, and 29 in the ischemia group displayed significant > or = 2.0-fold up-regulation in messenger RNA levels, and 185 transcripts in the APC group, 55 in the IPC group, and 49 in the ischemia group displayed significant > or = 2.0-fold down-regulation. Many of these transcripts were unknown genes. A high number of commonly regulated genes were found in IPC and APC (39 up-regulated, 17 down-regulated). Genes commonly regulated included those associated with cell defense (heat shock protein 10, aldose reductase, Bcl xS). Conversely, a pool of protective and antiprotective genes was differentially regulated in APC versus IPC (heat shock protein 27/70, programmed cell death 8), suggesting trigger-dependent transcriptome variability. CONCLUSIONS: The novel microarray technology provides evidence for distinct cardioprotective phenotypes in IPC and APC. The observed transcriptional changes raise the possibility of a second window of protection by volatile anesthetics. The authors' molecular portraits are the first global genomic comparison between IPC and APC. PMID- 15108960 TI - Isoflurane and sevoflurane precondition against neutrophil-induced contractile dysfunction in isolated rat hearts. AB - BACKGROUND: The authors tested the hypothesis that pretreatment with isoflurane or sevoflurane can protect the heart against neutrophil-induced contractile dysfunction. METHODS: Studies were conducted in buffer-perfused and paced isolated rat hearts. Left ventricular developed pressure served as an index of contractility. Pretreatment consisted of administration of 1.0 minimum alveolar concentration isoflurane or sevoflurane for 15 min followed by a 10-min washout and was performed in the absence and presence of the adenosine triphosphate sensitive potassium channel inhibitor glibenclamide (10 microM). Polymorphonuclear neutrophils and platelet-activating factor were then added to the perfusate for 10 min, followed by 30 min of recovery. Neutrophil retention was assessed from the difference between those administered and collected in coronary effluent and measurements of myeloperoxidase in myocardial samples. Isolated hearts were also used to assess the effect of volatile anesthetic pretreatment on cardiac dysfunction caused by enzymatically generated superoxide. In additional studies, the authors evaluated the effect of volatile anesthetic pretreatment on the adherence of neutrophils to isolated rat aortic segments. RESULTS: Platelet-activating factor-stimulated neutrophils caused marked and persistent reductions (> 50%) in left ventricular developed pressure. Pretreatment with either isoflurane or sevoflurane abolished these effects, as well as the associated increases in neutrophil retention. Glibenclamide did not alter these actions of the anesthetics. Pretreatment with either volatile anesthetic attenuated the reductions in left ventricular developed pressure caused by exogenous superoxide and abolished the increases in neutrophil adherence in the aortic segments. CONCLUSION: Isoflurane and sevoflurane preconditioned the heart against neutrophil-induced contractile dysfunction. This action was associated with an inhibition to neutrophil adherence and likely involved an increased resistance of the myocardium to oxidant-induced injury; the adenosine triphosphate-sensitive potassium channels played no apparent role. PMID- 15108961 TI - Attenuation of mitochondrial respiration by sevoflurane in isolated cardiac mitochondria is mediated in part by reactive oxygen species. AB - BACKGROUND: Anesthetic preconditioning protects against cardiac ischemia/reperfusion injury. Increases in reduced nicotinamide adenine dinucleotide and reactive oxygen species during sevoflurane exposure suggest attenuated mitochondrial electron transport as a trigger of anesthetic preconditioning. The authors investigated the effects of sevoflurane on respiration in isolated cardiac mitochondria. METHODS: Mitochondria were isolated from fresh guinea pig hearts, and mitochondrial oxygen consumption was measured in the presence of complex I (pyruvate) or complex II (succinate) substrates. The mitochondria were exposed to 0, 0.13, 0.39, 1.3, or 3.9 mM sevoflurane. State 3 respiration was determined after adenosine diphosphate addition. The reactive oxygen species scavengers manganese(III) tetrakis (4-benzoic acid) porphyrin chloride and N-tert-Butyl-a-(2-sulfophenyl)nitrone sodium (10 microM each), or the K(ATP) channel blockers glibenclamide (2 microM) or 5-hydroxydecanoate (300 microM), were given alone or before 1.3 mM sevoflurane. RESULTS: Sevoflurane attenuated respiration for both complex I and complex II substrates, depending on the dose. Glibenclamide and 5-hydroxydecanoate had no effect on this attenuation. Both scavengers, however, abolished the sevoflurane-induced attenuation for complex I substrates, but not for complex II substrates. CONCLUSION: The findings suggest that sevoflurane-induced attenuation of complex I is mediated by reactive oxygen species, whereas attenuation of other respiratory complexes is mediated by a different mechanism. The opening of mitochondrial K(ATP) channels by sevoflurane does not seem to be involved in this effect. Thus, reactive oxygen species formation may not only result from attenuated electron transport by sevoflurane, but it may also contribute to complex I attenuation, possibly leading to a positive feedback and amplification of sevoflurane-induced reactive oxygen species formation in triggering anesthetic preconditioning. PMID- 15108962 TI - Reactive oxygen species precede protein kinase C-delta activation independent of adenosine triphosphate-sensitive mitochondrial channel opening in sevoflurane induced cardioprotection. AB - BACKGROUND: In the current study, the authors investigated the distinct role and relative order of protein kinase C (PKC)-delta, adenosine triphosphate-sensitive mitochondrial K+ (mito K+(ATP)) channels, and reactive oxygen species (ROS) in the signal transduction of sevoflurane-induced cardioprotection and specifically addressed their mechanistic link. METHODS: Isolated rat trabeculae were preconditioned with 3.8% sevoflurane and subsequently subjected to an ischemic protocol by superfusion of trabeculae with hypoxic, glucose-free buffer (40 min) followed by 60 min of reperfusion. In addition, the acute affect of sevoflurane on PKC-delta and PKC-epsilon translocation and nitrotyrosine formation was established with use of immunofluorescent analysis. The inhibitors chelerythrine (6 microM), rottlerin (1 microM), 5-hydroxydecanoic acid sodium (100 microM), and n-(2-mercaptopropionyl)-glycine (300 microM) were used to study the particular role of PKC, PKC-delta, mito K+(ATP), and ROS in sevoflurane-related intracellular signaling. RESULTS: Preconditioning of trabeculae with sevoflurane preserved contractile function after ischemia. This contractile preservation was dependent on PKC-delta activation, mito K+(ATP) channel opening, and ROS production. In addition, on acute stimulation by sevoflurane, PKC-delta but not PKC-epsilon translocated to the sarcolemmal membrane. This translocation was inhibited by PKC inhibitors and ROS scavenging but not by inhibition of mito K+(ATP) channels. Furthermore, sevoflurane directly induced nitrosylation of sarcolemmal proteins, suggesting the formation of peroxynitrite. CONCLUSIONS: In sevoflurane-induced cardioprotection, ROS release but not mito K+(ATP) channel opening precedes PKC-delta activation. Sevoflurane induces sarcolemmal nitrotyrosine formation, which might be involved in the recruitment of PKC-delta to the cell membrane. PMID- 15108963 TI - Adenosine and a nitric oxide donor enhances cardioprotection by preconditioning with isoflurane through mitochondrial adenosine triphosphate-sensitive K+ channel dependent and -independent mechanisms. AB - BACKGROUND: Preconditioning with isoflurane has been shown to confer cardioprotection via activation of mitochondrial adenosine triphosphate-sensitive K+ (mito K(ATP)) channels. However, the relative contribution of mito K(ATP) channel and non-mito K(ATP) channel mechanisms to isoflurane-mediated cardioprotection has not been investigated. METHODS: Isolated and buffer-perfused rat hearts were used. Flavoprotein fluorescence was monitored as an index for mito K(ATP) channel activity. Isovolumic left ventricular function and infarct size were measured as indices for cardioprotection. RESULTS: Flavoprotein fluorescence, which was monitored as an index for mito K(ATP) channel activity, was increased by isoflurane and a known mito K(ATP) channel opener, diazoxide, in a 5-hydroxydecanoate-sensitive manner. Although flavoprotein oxidation induced by diazoxide was dissipated soon after its removal from the buffer, flavoprotein oxidation induced by isoflurane was sustained after cessation of the treatment. The sustained increase in flavoprotein oxidation was associated with a significant reduction in infarct size after 30 min of ischemia followed by 120 min of reperfusion. Although adenosine and S-nitroso-N-acetyl-penicillamine each alone did not increase flavoprotein fluorescence, nor did they confer significant cardioprotection, coadministration of adenosine and S-nitroso-N-acetyl penicillamine with isoflurane conferred a highly significant reduction of infarct size and improvement of left ventricular function without increasing flavoprotein oxidation over isoflurane alone. The early treatment with 5-hydroxydecanoate before and during preconditioning completely reversed flavoprotein oxidation and inhibited the infarct-sparing effect of isoflurane and combined preconditioning with isoflurane, adenosine, and S-nitroso-N-acetyl-penicillamine. The late treatment with 5-hydroxydecanoate after preconditioning abolished flavoprotein oxidation and the infarct-sparing effect of isoflurane but only partially inhibited cardioprotection conferred by the combined preconditioning, despite complete abrogation of flavoprotein oxidation. CONCLUSIONS: Mito K(ATP) channel activation is the essential trigger of both preconditioning with isoflurane and combined preconditioning with isoflurane, adenosine, and S-nitroso-N-acetyl penicillamine. Mito K(ATP) channel activation is also a crucial mediator of cardioprotection afforded by preconditioning with isoflurane. However, enhanced cardioprotection conferred by combined preconditioning is mediated through both mito K(ATP) channel-dependent and -independent mechanisms. PMID- 15108964 TI - Isoflurane produces delayed preconditioning against myocardial ischemia and reperfusion injury: role of cyclooxygenase-2. AB - BACKGROUND: Whether volatile anesthetics produce a second window of preconditioning is unclear. The authors tested the hypothesis that isoflurane causes delayed preconditioning against infarction and, further, that cyclooxygenase (COX)-2 mediates this beneficial effect. METHODS: Rabbits (n = 43) were randomly assigned to receive 0.9% intravenous saline, the selective COX-2 inhibitor celecoxib (3 mg/kg intraperitoneal) five times over 2 days before coronary artery occlusion and reperfusion, or isoflurane (1.0 minimum alveolar concentration) 24 h before acute experimentation in the absence or presence of celecoxib pretreatment. Two additional groups of rabbits received a single dose of celecoxib either 30 min before or 21.5 h after administration of isoflurane. Rabbits were then instrumented for measurement of hemodynamics and underwent 30 min of coronary occlusion followed by 3 h of reperfusion. Myocardial infarct size was measured using triphenyltetrazolium staining. Western immunoblotting to examine COX-1 and COX-2 protein expression was performed in rabbit hearts that had or had not been exposed to isoflurane. RESULTS: Isoflurane significantly (P < 0.05) reduced infarct size (22 +/- 3% of the left ventricular area at risk) as compared with control (39 +/- 2%). Celecoxib alone had no effect on infarct size (36 +/- 4%) but abolished isoflurane-induced cardioprotection (36 +/- 4%). A single dose of celecoxib administered 2.5 h before coronary occlusion and reperfusion also abolished the delayed protective effects of isoflurane (36 +/- 4%), but celecoxib given 30 min before exposure to isoflurane had no effect (22 +/- 4%). Isoflurane did not alter COX-1 and COX-2 protein expression. CONCLUSIONS: The results indicate that the volatile anesthetic isoflurane produces a second window of preconditioning against myocardial ischemia and reperfusion injury. Furthermore, COX-2 is an important mediator of isoflurane induced delayed preconditioning. PMID- 15108965 TI - Protein kinase C translocation and Src protein tyrosine kinase activation mediate isoflurane-induced preconditioning in vivo: potential downstream targets of mitochondrial adenosine triphosphate-sensitive potassium channels and reactive oxygen species. AB - BACKGROUND: The authors tested the hypotheses that protein kinase C (PKC) specific isoform translocation and Src protein tyrosine kinase (PTK) activation play important roles in isoflurane-induced preconditioning in vivo. METHODS: Rats (n = 125) instrumented for measurement of hemodynamics underwent 30 min of coronary artery occlusion followed by 2 h of reperfusion and received 0.9% saline (control); PKC inhibitors chelerythrine (5 mg/kg), rottlerin (0.3 mg/kg), or PKC epsilonV1-2 peptide (1 mg/kg); PTK inhibitors lavendustin A (1 mg/kg) or 4-amino 5-(4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP1; 1 mg/kg); mitochondrial adenosine triphosphate-sensitive potassium channel antagonist 5 hydroxydecanote (10 mg/kg); or reactive oxygen species scavenger N-acetylcysteine (150 mg/kg) in the absence and presence of a 30-min exposure to isoflurane (1.0 minimum alveolar concentration) in separate groups. Isoflurane was discontinued 15 min before coronary occlusion (memory period). Infarct size was determined using triphenyltetrazolium staining. Immunohistochemistry and confocal microscopic imaging were performed to examine PKC translocation in separate groups of rats. RESULTS: Isoflurane significantly (P < 0.05) reduced infarct size (40 +/- 3% [n = 13]) as compared with control experiments (58 +/- 2% [n = 12]). Chelerythrine, rottlerin, PKC-epsilonV1-2 peptide, lavendustin A, PP1, 5 hydroxydecanote, and N-acetylcysteine abolished the anti-ischemic actions of isoflurane (58 +/- 2% [n = 8], 50 +/- 3% [n = 9], 53 +/- 2% [n = 9], 59 +/- 3% [n = 6], 57 +/- 3% [n = 7], 60 +/- 3% [n = 7], and 53 +/- 3% [n = 6], respectively). Isoflurane stimulated translocation of the delta and epsilon isoforms of PKC to sarcolemmal and mitochondrial membranes, respectively. CONCLUSIONS: Protein kinase C-delta, PKC-epsilon, and Src PTK mediate isoflurane-induced preconditioning in the intact rat heart. Opening of mitochondrial adenosine triphosphate-sensitive potassium channels and generation of reactive oxygen species are upstream events of PKC activation in this signal transduction process. PMID- 15108966 TI - Nuclear factor kappaB and anesthetic preconditioning during myocardial ischemia reperfusion. AB - BACKGROUND: Volatile anesthetic preconditioning (APC) protects against myocardial ischemia-reperfusion (IR) injury, but the precise mechanisms underlying this phenomenon remain undefined. To investigate the molecular mechanism of APC in myocardial protection, the activation of nuclear factor (NF) kappaB and its regulated inflammatory mediators expression were examined in the current study. METHODS: Hearts from male rats were isolated, Langendorff perfused, and randomly assigned to one of three groups: (1) the control group: hearts were continuously perfused for 130 min; (2) the IR group: 30 min of equilibration, 15 min of baseline, 25 min of ischemia, 60 min of reperfusion; and (3) the APC + IR group: 30 min of equilibration, 10 min of sevoflurane exposure and a 5-min washout, 25 min of global ischemia, 60 min of reperfusion. Tissue samples were acquired at the end of reperfusion. NF-kappaB activity was determined by electrophoretic mobility shift assay. The NF-kappaB inhibitor, IkappaB-alpha, was determined by Western blot analysis. Myocardial inflammatory mediators, including tumor necrosis factor alpha, interleukin 1, intercellular adhesion molecule 1, and inducible nitric oxide synthase, were also assessed by Western blot analysis. RESULTS: Nuclear factor kappaB-DNA binding activity was significantly increased at the end of reperfusion in rat myocardium, and cytosolic IkappaB-alpha was decreased. Supershift assay revealed the involvement of NF-kappaB p65 and p50 subunits. APC with sevoflurane attenuated NF-kappaB activation and reduced the expression of tumor necrosis factor alpha, interleukin 1, intercellular adhesion molecule 1, and inducible nitric oxide synthase. APC also reduced infarct size and creatine kinase release and improved myocardial left ventricular developed pressure during IR. CONCLUSIONS: The results of this study indicate that attenuation of NF-kappaB activation and subsequent down-regulation of NF-kappaB dependent inflammatory gene expression plays an important role in the protective mechanism of APC against acute myocardial IR injury. PMID- 15108967 TI - Cyclooxygenase-2 mediates ischemic, anesthetic, and pharmacologic preconditioning in vivo. AB - BACKGROUND: Cyclooxygenase-2 (COX-2) mediates the late phase of ischemic preconditioning (IPC), but whether this enzyme modulates early IPC, anesthetic induced preconditioning (APC), or other forms of pharmacologic preconditioning (PPC) is unknown. The authors tested the hypothesis that COX-2 is an essential mediator of IPC, APC, and PPC in vivo. METHODS: Barbiturate-anesthetized dogs (n = 91) were instrumented for measurement of hemodynamics and randomly assigned to receive IPC (four 5-min coronary occlusions interspersed with 5-min reperfusions), APC (1.0 minimum alveolar concentration of isoflurane for 30 min), or PPC (selective mitochondrial K(ATP) channel opener diazoxide, 2.5 mg/kg intravenous) in the presence or absence of pretreatment with oral aspirin (650 mg), the selective COX-2 inhibitor celecoxib (200 mg), or acetaminophen (500 mg) administered 24, 12, and 2 h before experimentation in 12 separate experimental groups. All dogs were subjected to a 60-min coronary artery occlusion followed by 3 h of reperfusion. Myocardial infarct size and coronary collateral blood flow were quantified with triphenyltetrazolium staining and radioactive microspheres, respectively. Myocardial 6-keto-prostaglandin F1alpha, a stable metabolite of prostacyclin, was measured (enzyme immunoassay) in separate experiments (n = 8) before and after isoflurane administration, in the presence or absence of celecoxib. RESULTS: No significant differences in baseline hemodynamics or the left ventricular area at risk for infarction were observed between groups. IPC, isoflurane, and diazoxide all decreased myocardial infarct size (9 +/- 1, 12 +/- 2, and 11 +/- 1%, respectively) as compared with control (30 +/- 1%). Celecoxib alone had no effect on infarct size (26 +/- 3%) but abolished IPC (30 +/-3%), APC (30 +/- 3%), and PPC (26 +/- 1%). Aspirin (24 +/- 3%) and acetaminophen alone (29 +/- 2%) did not alter infarct size or abolish APC-induced protection (18 +/- 1 and 19 +/- 1%, respectively). Isoflurane increased myocardial 6-keto prostaglandin F1alpha to 463 +/- 267% of baseline in the absence but not in the presence (94 +/- 13%) of celecoxib. CONCLUSIONS: The results indicate that COX-2 is a critical mediator of IPC, APC, and PPC in dogs. The role of cyclooxygenase enzymes as obligatory mediators of myocardial protection produced by diverse preconditioning stimuli may have implications for the clinical use of COX-2 inhibitors. PMID- 15108968 TI - Role of tyrosine kinase in desflurane-induced preconditioning. AB - BACKGROUND: Short administration of volatile anesthetics preconditions myocardium and protects the heart against the consequences of subsequent ischemia. Activation of tyrosine kinase is implicated in ischemic preconditioning. The authors investigated whether desflurane-induced preconditioning depends on activation of tyrosine kinase. METHODS: Sixty-four rabbits were instrumented for measurement of left ventricular pressure, cardiac output, and myocardial infarct size (IS). All rabbits were subjected to 30 min of occlusion of a major coronary artery and 2 h of subsequent reperfusion. Rabbits underwent a treatment period consisting of either no intervention for 35 min (control group, n = 12) or 15 min of 1 minimum alveolar concentration desflurane inhalation followed by a 10-min washout period (desflurane group, n = 12). Four additional groups received the tyrosine kinase inhibitor genistein (5 mg/kg) or lavendustin A (1.3 mg/kg) at the beginning of the treatment period with (desflurane-genistein group, n = 11; desflurane-lavendustin A group, n = 12) or without desflurane inhalation (genistein group, n = 9; lavendustin A group, n = 8). RESULTS: Hemodynamic values were similar in all groups during baseline (left ventricular pressure, 87 +/- 14 mmHg (mean +/- SD]; cardiac output, 198 +/- 47 ml/min), during coronary artery occlusion (left ventricular pressure, 78 +/- 12 mmHg; cardiac output, 173 +/- 39 ml/min), and after 2 h of reperfusion (left ventricular pressure, 59 +/- 17; cardiac output, 154 +/- 43 ml/min). IS in the control group was 55 +/- 10% of the area at risk. The tyrosine inhibitors had no effect on IS (genistein group, 56 +/ 13%; lavendustin A group, 49 +/- 13%; each P = 1.0 vs. control group). Desflurane preconditioning reduced IS to 40 +/- 15% (P = 0.04 vs. control group). Tyrosine kinase inhibitor administration had no effect on IS reduction (desflurane-genistein group, 44 +/- 13%; desflurane-lavendustin A group, 44 +/- 16%; each P = 1.0 vs. desflurane group). CONCLUSION: Desflurane-induced preconditioning does not depend on tyrosine kinase activation. PMID- 15108969 TI - Morphine preconditions Purkinje cells against cell death under in vitro simulated ischemia-reperfusion conditions. AB - BACKGROUND: Morphine pretreatment via activation of delta1-opioid receptors induces cardioprotection. In this study, the authors determined whether morphine preconditioning induces ischemic tolerance in neurons. METHODS: Cerebellar brain slices from adult Sprague-Dawley rats were incubated with morphine at 0.1-10 microM in the presence or absence of various antagonists for 30 min. They were then kept in morphine- and antagonist-free buffer for 30 min before they were subjected to simulated ischemia (oxygen-glucose deprivation) for 20 min. After being recovered in oxygenated artificial cerebrospinal fluid for 5 h, they were fixed for morphologic examination to determine the percentage of undamaged Purkinje cells. RESULTS: The survival rate of Purkinje cells was significantly higher in slices preconditioned with morphine (> or = 0.3 microM) before the oxygen-glucose deprivation (57 +/- 4% at 0.3 microM morphine) than that of the oxygen-glucose deprivation alone (39 +/- 3%, P < 0.05). This morphine preconditioning-induced neuroprotection was abolished by naloxone, a non-type selective opioid receptor antagonist, by naltrindole, a selective delta-opioid receptor antagonist, or by 7-benzylidenenaltrexone, a selective delta1-opioid receptor antagonist. However, the effects were not blocked by the mu-, kappa-, or delta2-opioid receptor antagonists, beta-funaltrexamine, nor-binaltorphimine, or naltriben, respectively. Morphine preconditioning-induced neuroprotection was partially blocked by the selective mitochondrial adenosine triphosphate-sensitive potassium channel antagonist, 5-hydroxydecanoate, or the mitochondrial electron transport inhibitor, myxothiazol. None of the inhibitors used in this study alone affected the simulated ischemia-induced neuronal death. CONCLUSIONS: These data suggest that morphine preconditioning is neuroprotective. This neuroprotection may be delta1-opioid receptor dependent and may involve mitochondrial adenosine triphosphate-sensitive potassium channel activation and free radical production. Because morphine is a commonly used analgesic, morphine preconditioning may be explored further for potential clinical use to reduce ischemic brain injury. PMID- 15108970 TI - Dual exposure to sevoflurane improves anesthetic preconditioning in intact hearts. AB - BACKGROUND: Anesthetic preconditioning (APC) with sevoflurane reduces myocardial ischemia-reperfusion injury. The authors tested whether two brief exposures to sevoflurane would lead to a better preconditioning state than would a single longer exposure and whether dual exposure to a lower (L) concentration of sevoflurane would achieve an outcome similar to that associated with a single exposure to a higher (H) concentration. METHODS: Langendorff-prepared guinea pig hearts were exposed to 0.4 mM sevoflurane once for 15 min (H1-15; n = 8) or 0.4 mM (H2-5; n = 8) or 0.2 mM sevoflurane (L2-5; n = 8) twice for 5 min, with a 5 min washout period interspersed. Sevoflurane was then washed out for 20 min before 30 min of global no-flow ischemia and 120 min of reperfusion. Control hearts (n = 8) were not subjected to APC. Left ventricular pressure was measured isovolumetrically. Ventricular infarct size was determined by tetrazolium staining and cumulative planimetry. Values are expressed as mean +/- SD. RESULTS: The authors found a better functional return and a lesser percentage of infarction on reperfusion in H2-5 (28 +/- 9%) than in H1-15 (36 +/- 8%; P < 0.05), L2-5 (43 +/- 6%; P < 0.05), or control hearts (52 +/- 7%; P < 0.05). CONCLUSION: These results suggest that APC depends not only on the concentration but also on the protocol used for preconditioning. Similarly to ischemic preconditioning, repeated application of the volatile anesthetic seems to be more important than the duration of exposure in initiating the signaling sequence that elicits APC at clinically relevant concentrations. Therefore, repeated cycles of anesthetic exposure followed by volatile anesthetic-free periods may be beneficial for APC in the clinical setting. PMID- 15108971 TI - Contribution of reactive oxygen species to isoflurane-induced sensitization of cardiac sarcolemmal adenosine triphosphate-sensitive potassium channel to pinacidil. AB - BACKGROUND: Myocardial protection by volatile anesthetics involves activation of cardiac adenosine triphosphate-sensitive potassium (K(ATP)) channels. The authors have previously shown that isoflurane enhances sensitivity of the sarcolemmal K(ATP) channel to the opener, pinacidil. Because reactive oxygen species seem to be mediators in anesthetic preconditioning, the authors investigated whether they contribute to the mechanism of the sensitization effect by isoflurane. METHODS: Ventricular myocytes were isolated from guinea pig hearts for the whole cell patch clamp recordings of the sarcolemmal K(ATP) channel current (I(KAPT)). Free radical scavengers N-acetyl-L-cysteine, carnosine, superoxide dismutase, and catalase were used to investigate whether reactive oxygen species mediate isoflurane facilitation of the channel opening by pinacidil. A possible role of the mitochondrial K(ATP) channels was tested using a blocker of these channels, 5 hydroxydecanoate. RESULTS: The mean density (+/- SEM) of I(KAPT) elicited by pinacidil (20 microM) was 18.9 +/- 1.8 pA/pF (n = 11). In the presence of isoflurane (0.55 mM), the density of pinacidil-activated I(KAPT) increased to 38.5 +/- 2.4 pA/pF (n = 9). Concurrent application of isoflurane and N-acetyl-L cysteine decreased the sensitization effect by isoflurane in a concentration dependent manner, whereby the densities of I(KAPT) were 32.6 +/- 1.4 (n = 6), 26.2 +/- 2.3 (n = 6), and 19.4 +/- 2.1 pA/pF (n = 8) at 100, 250, and 500 microM N-acetyl-L-cysteine, respectively. Concurrent application of isoflurane and carnosine (100 microM), superoxide dismutase (100 U/ml), or catalase (100 U/ml) attenuated the densities of I(KAPT) to 27.9 +/- 2.6, 27.2 +/- 2.9, and 25.9 +/- 2.2 pA/pF, respectively. None of the scavengers affected activation of I(KAPT) by pinacidil alone. 5-Hydroxydecanoate (100 microM) did not alter the sensitization effect by isoflurane, and the density of I(KAPT) in this group was 37.1 +/- 3.8 pA/pF (n= 6). CONCLUSION: These results suggest that reactive oxygen species contribute to the mechanism by which isoflurane sensitizes the cardiac sarcolemmal K(ATP) channel to the opener, pinacidil. PMID- 15108972 TI - Desflurane-induced preconditioning alters calcium-induced mitochondrial permeability transition. AB - BACKGROUND: Recent investigations have focused on the pivotal role of the mitochondria in the underlying mechanisms volatile anesthetic-induced myocardial preconditioning. This study aimed at examining the effect of anesthetic preconditioning on mitochondrial permeability transition (MPT) pore opening. METHODS: Anesthetized open chest rabbits were randomized to one of four groups and underwent 10 min of ischemia, except for the sham 1 group (n = 12). Before this, they underwent a treatment period consisting of (1) no intervention (ischemic group; n = 12), (2) 30 min of desflurane inhalation (8.9% end-tidal concentration) followed by a 15-min washout period (desflurane group; n = 12), or (3) ischemic preconditioning (IPC group; n = 12). A second set of experiments was performed to evaluate the effect of a putative mitochondrial adenosine triphosphate-sensitive potassium channel antagonist, 5-hydroxydecanoate (5-HD). The animals underwent the same protocol as previously, plus pretreatment with 5 mg/kg 5-HD. They were randomized to one of five groups: the sham 2 group, receiving no 5-HD (n = 12); the sham 5-HD group (n = 12); the ischemic 5-HD group (n = 12), the desflurane 5-HD group (n = 12), and the IPC 5-HD group (n = 12). At the end of the protocol, the hearts were excised, and mitochondria were isolated. MPT pore opening was assessed by measuring the amount of calcium required to trigger a massive calcium release indicative of MPT pore opening. RESULTS: Desflurane and IPC group mitochondria needed a higher calcium load than ischemic group mitochondria (362 +/- 84, 372 +/- 74, and 268 +/- 110 microM calcium, respectively; P < 0.05) to induce MPT pore opening. The sham 1 and sham 2 groups needed a similar amount of calcium to trigger mitochondrial calcium release (472 +/- 70 and 458 +/- 90 microM calcium, respectively). 5-HD preadministration had no effect on sham animals (458 +/- 90 and 440 +/- 128 microM calcium without and with 5-HD, respectively) and ischemic group animals (268 +/- 110 and 292 +/- 102 microM calcium without and with 5-HD, respectively) but abolished the effects of desflurane on calcium-induced MPT pore opening (362 +/- 84 microM calcium without 5-HD vs. 238 +/- 96 microM calcium with 5-HD; P < 0.05) and IPC (372 +/- 74 microM calcium without 5-HD vs. 270 +/- 104 microM calcium with 5-HD; P < 0.05). CONCLUSION: Like ischemic preconditioning, desflurane improved the resistance of the transition pore to calcium-induced opening. This effect was inhibited by 5 HD, suggesting a link between mitochondrial adenosine triphosphate-sensitive potassium and MPT. PMID- 15108973 TI - Reduced efficacy of volatile anesthetic preconditioning with advanced age in isolated rat myocardium. AB - BACKGROUND: Ischemic preconditioning and anesthetic preconditioning (APC) are reported to decrease myocardial infarct size during ischemia-reperfusion injury. However, the beneficial effects of ischemic preconditioning have been shown to decrease with advancing age. Although the mechanisms of ischemic preconditioning and APC are thought to be similar, it is not known whether the beneficial effects of APC are also reduced in the aged myocardium. METHODS: Male Fischer 344 rats of three age groups (2-4, 10-12, and 20-24 months) were used. Hearts were Langendorff perfused. Six hearts in each age group were pretreated with 10 min of sevoflurane and a 5-min washout before 25 min of ischemia and 60 min of reperfusion. Six control hearts in each age group received no treatment before ischemia. Nuclear magnetic resonance was used to measure intracellular Na, intracellular Ca, and intracellular pH, respectively. Left ventricular developed pressure, creatine kinase, and infarct size were measured. RESULTS: Ischemia decreases intracellular pH and increases intracellular Na and intracellular Ca in all age groups. APC blunts the pH decreases in young adult and middle-aged rats, but not in aged rats. APC decreased intracellular Na and intracellular Ca accumulation during ischemia in young adult and middle-aged hearts. APC improved adenosine triphosphate recovery in young rats but not in aged rats. Creatine kinase and infarct sizes were significantly reduced and left ventricular developed pressure was improved with APC in the young adult and middle-aged groups but not the aged group. CONCLUSIONS: The benefits of APC are significantly reduced with advanced age in an isolated rat heart model. PMID- 15108974 TI - Mandibular advancement improves the laryngeal view during direct laryngoscopy performed by inexperienced physicians. AB - BACKGROUND: When oral or nasal fiberoptic laryngoscopy is attempted, mandibular advancement has been reported to improve the laryngeal view. The authors hypothesized that mandibular advancement may also improve the laryngeal view during direct laryngoscopy. METHODS: Forty patients undergoing elective surgery under general anesthesia were included in this study. After establishment of an adequate level of anesthesia and muscle relaxation, direct laryngoscopy was performed by inexperienced physicians. Four different maneuvers--simple direct laryngoscopy without any assistance (C), simple direct laryngoscopy with mandibular advancement (M), simple direct laryngoscopy with the BURP maneuver (backward, upward, rightward pressure maneuver of the larynx; B), and simple direct laryngoscopy with both mandibular advancement and the BURP maneuver (BM)- were attempted in each subject, and the laryngeal aperture was videotaped with each procedure. An instructor in anesthesiology who was blinded to the procedure evaluated the visualization by reviewing videotape off-line, using the Cormack Lehane classification system (grades I-IV) and a rating score within each subject (1 = best view; 4 = poorest view). The Friedman test followed by the Student Newman-Keuls test was performed for statistical comparison. P < 0.05 was considered significant. RESULTS: The laryngeal view was improved with M and B when compared with C (P < 0.05 by both rating and Cormack-Lehane evaluation). BM was the most effective method to visualize the laryngeal aperture (P < 0.05, vs. B and M by rating evaluation), whereas B and M were the second and the third most effective methods, respectively. No statistical difference was observed between B and M with the Cormack-Lehane classification. CONCLUSION: Mandibular advancement improves the laryngeal view during direct laryngoscopy performed by inexperienced physicians. PMID- 15108975 TI - Remifentanil induces systemic arterial vasodilation in humans with a total artificial heart. AB - BACKGROUND: To assess intrinsic vascular effects of remifentanil, increased concentrations were infused in critically ill patients with a total artificial heart. METHODS: In the early postoperative period after implantation of a total artificial heart, nine ventilated patients requiring short general anesthesia were included in this study. After anesthesia was induced with 0.3 mg/kg intravenous etomidate, the artificial heart settings were modified to render cardiac output "preload-independent." While maintenance of anesthesia was ensured by a continuous infusion of etomidate, increased concentrations of remifentanil (from 0.1 to 1 microg x kg(-1) x min(-1)) were infused in steps of 5 min under hemodynamic monitoring, including left and right atrial pressures, systemic and pulmonary arterial pressures, and left and right cardiac indices. The invasive procedure was started under the highest concentration of remifentanil tolerated by the patient. Infusion of remifentanil was stopped at the end of the invasive procedure, while etomidate infusion was maintained. New hemodynamic measurements were performed at the end of the 12-min recovery period. RESULTS: Remifentanil produced a dose-dependent and significant decrease in systemic arterial pressure and vascular resistances (n = 9) from a concentration of 0.25 microg x kg(-1) x min(-1). No significant changes were observed on pulmonary vascular resistances (n = 6). Neither right (n = 9) nor left (n = 6) atrial pressures were affected by remifentanil infusion. Hemodynamic variables returned to baseline value over the 12-min recovery period. CONCLUSIONS: In humans with a total artificial heart, remifentanil induces a systemic arterial vasodilation without significant effect on the capacitance vessels. PMID- 15108976 TI - Variable ventilation improves perioperative lung function in patients undergoing abdominal aortic aneurysmectomy. AB - BACKGROUND: Optimizing perioperative mechanical ventilation remains a significant clinical challenge. Experimental models indicate that "noisy" or variable ventilation (VV)--return of physiologic variability to respiratory rate and tidal volume--improves lung function compared with monotonous control mode ventilation (CV). VV was compared with CV in patients undergoing abdominal aortic aneurysmectomy, a patient group known to be at risk of deteriorating lung function perioperatively. METHODS: After baseline measurements under general anesthesia (CV with a tidal volume of 10 ml/kg and a respiratory rate of 10 breaths/min), patients were randomized to continue CV or switch to VV (computer control of the ventilator at the same minute ventilation but with 376 combinations of respiratory rate and tidal volume). Lung function was measured hourly for the next 6 h during surgery and recovery. RESULTS: Forty-one patients for aneurysmectomy were studied. The characteristics of the patients in the two groups were similar. Repeated-measures analysis of variance (group x time interaction) revealed greater arterial oxygen partial pressure (P = 0.011), lower arterial carbon dioxide partial pressure (P = 0.012), lower dead space ventilation (P = 0.011), increased compliance (P = 0.049), and lower mean peak inspiratory pressure (P = 0.013) with VV. CONCLUSIONS: The VV mode of ventilation significantly improved lung function over CV in patients undergoing abdominal aortic aneurysmectomy. PMID- 15108977 TI - Sequential effects of increasing propofol sedation on frontal and temporal cortices as indexed by auditory event-related potentials. AB - BACKGROUND: It is an open question whether cognitive processes of auditory perception that are mediated by functionally different cortices exhibit the same sensitivity to sedation. The auditory event-related potentials P1, mismatch negativity (MMN), and early right anterior negativity (ERAN) originate from different cortical areas and reflect different stages of auditory processing. The P1 originates mainly from the primary auditory cortex. The MMN is generated in or in the close vicinity of the primary auditory cortex but is also dependent on frontal sources. The ERAN mainly originates from frontal generators. The purpose of the study was to investigate the effects of increasing propofol sedation on different stages of auditory processing as reflected in P1, MMN, and ERAN. METHODS: The P1, the MMN, and the ERAN were recorded preoperatively in 18 patients during four levels of anesthesia adjusted with target-controlled infusion: awake state (target concentration of propofol 0.0 microg/ml), light sedation (0.5 microg/ml), deep sedation (1.5 microg/ml), and unconsciousness (2.5 3.0 microg/ml). Simultaneously, propofol anesthesia was assessed using the Bispectral Index. RESULTS: Propofol sedation resulted in a progressive decrease in amplitudes and an increase of latencies with a similar pattern for MMN and ERAN. MMN and ERAN were elicited during sedation but were abolished during unconsciousness. In contrast, the amplitude of the P1 was unchanged by sedation but markedly decreased during unconsciousness. CONCLUSION: The results indicate differential effects of propofol sedation on cognitive functions that involve mainly the auditory cortices and cognitive functions that involve the frontal cortices. PMID- 15108978 TI - Influence of chronic phenytoin administration on the pharmacokinetics and pharmacodynamics of vecuronium. AB - BACKGROUND: The duration of action of vecuronium is reduced in patients receiving phenytoin. In this study, the authors examined, simultaneously, the influence of phenytoin on both the pharmacokinetics and the pharmacodynamics of vecuronium. METHODS: This study was approved by the institutional review board of the University of California, San Francisco, and patients gave written informed consent. Twenty-two patients, 11 taking phenytoin and all scheduled to undergo prolonged neurosurgical procedures with general anesthesia, participated in the study. In 12 patients (6 phenytoin, 6 control), vecuronium was infused at 7.5 microg x kg(-1) x min(-1) until the first response (T1) of each train-of-four decreased by 50%; in the remaining 10 patients (5 phenytoin, 5 control), 200 microg/kg vecuronium was infused over 10 min. Arterial blood samples were drawn at intervals over the next 5-7 h. Plasma concentrations of vecuronium and 3 desacetylvecuronium were measured by capillary gas chromatography. Pharmacokinetic and pharmacodynamic modeling was used to characterize the disposition of vecuronium and patient responses to it in the two groups. RESULTS: Clearance was typically increased by 138% (95% confidence interval, 93-183%) in patients taking phenytoin. The effect of vecuronium was well described using a sigmoid Emax model. The concentration of vecuronium giving 50% twitch depression was increased 124% (45-202%) in patients taking phenytoin. CONCLUSIONS: Chronic phenytoin therapy reduces the effect of vecuronium by mechanisms that include both increased vecuronium metabolism and reduced sensitivity of the patient to circulating concentrations of vecuronium. PMID- 15108980 TI - Performance evaluation of two published closed-loop control systems using bispectral index monitoring: a simulation study. AB - BACKGROUND: Although automated closed-loop control systems may improve quality of care, their safety must be proved under extreme control conditions. This study describes a simulation methodology to test automated controllers and its application in a comparison of two published controllers for Bispectral Index (BIS)-guided propofol administration. METHODS: A patient simulator was developed to compare controllers. Using input scripts to dictate patient characteristics, target BIS values, and the time course of surgical events, the simulator continuously monitors the infusion pump under control and generates BIS values as a composite of modeled response to drug, perceived stimulation, and random noise. The simulator formats the output stream of BIS data as input to the controller under test to emulate the serial output of the actual BIS monitor. A published model-based controller and a classic proportional integral derivative controller were compared when using the BIS value as a controlled variable. Each controller was tested using a set of 10 virtual patients undergoing a fixed surgical profile that was repeated with BIS targets set at 30, 50, and 70. Controller performance was assessed using median (absolute) prediction error, divergence, wobble, and percentage time within BIS target range metrics. RESULTS: The median prediction error was significantly smaller for the proportional integral derivative controller than for the model-based controller. The median absolute prediction error was smaller for the model-based controller than for the proportional integral derivative controller for each BIS target, reaching statistical significance for targets 30 and 50. CONCLUSIONS: When simulating closed-loop control of BIS using propofol, the use of a patient-individualized, model-based adaptive closed-loop system with effect site control resulted in better control of BIS compared with a standard proportional integral derivative controller with plasma site control. Even under extreme conditions, the modeled-based controller exhibited no behavioral problems. PMID- 15108979 TI - Effect of amino acid infusion on central thermoregulatory control in humans. AB - BACKGROUND: Administration of protein or amino acids enhances thermogenesis, presumably by stimulating oxidative metabolism. However, hyperthermia results even when thermoregulatory responses are intact, suggesting that amino acids also alter central thermoregulatory control. Therefore, the authors tested the hypothesis that amino acid infusion increases the thermoregulatory set point. METHODS: Nine male volunteers each participated on 4 study days in randomized order: (1) intravenous amino acids infused at 4 kJ x kg(-1) x h(-1) for 2.5 h combined with skin-surface warming, (2) amino acid infusion combined with cutaneous cooling, (3) saline infusion combined with skin-surface warming, and (4) saline infusion combined with cutaneous cooling. RESULTS: Amino acid infusion increased resting core temperature by 0.3 +/- 0.1 degrees C (mean +/- SD) and oxygen consumption by 18 +/- 12%. Furthermore, amino acid infusion increased the calculated core temperature threshold (triggering core temperature at a designated mean skin temperature of 34 degrees C) for active cutaneous vasodilation by 0.3 +/- 0.3 degrees C, for sweating by 0.2 +/- 0.2 degrees C, for thermoregulatory vasoconstriction by 0.3 +/- 0.3 degrees C, and for thermogenesis by 0.4 +/- 0.5 degrees C. Amino acid infusion did not alter the incremental response intensity (i.e., gain) of thermoregulatory defenses. CONCLUSIONS: Amino acid infusion increased the metabolic rate and the resting core temperature. However, amino acids also produced a synchronous increase in all major autonomic thermoregulatory defense thresholds; the increase in core temperature was identical to the set point increase, even in a cold environment with amble potential to dissipate heat. In subjects with intact thermoregulatory defenses, amino acid-induced hyperthermia seems to result from an increased set point rather than increased metabolic rate per se. PMID- 15108981 TI - Effect of propofol on hypotonic swelling-induced membrane depolarization in human coronary artery smooth muscle cells. AB - BACKGROUND: Stretch (mechanical stress)-induced membrane depolarization of smooth muscle may contribute to basal vascular tone and myogenic control. Propofol induces vasodilation and inhibits myogenic control. Hypotonic swelling was used as a model of mechanical stress. The authors investigated the effects of propofol and 5-nitro-2-(3-phenylpropylamino)benzoic acid, a chloride channel and nonselective cation channel blocker, on hypotonicity-induced membrane depolarization in cultured human coronary artery smooth muscle cells. METHODS: A voltage-sensitive fluorescent dye, bis-(1,3-diethylthiobarbiturate)trimethine oxonol, was used to assess relative changes in membrane potential semiquantitatively. The cells were continuously perfused with Earle's balanced salt solution containing 200 nM bis-(1,3-diethylthiobarbiturate)trimethine oxonol and exposed sequentially to isotonic and hypotonic medium. In a second series of experiments, the cells were exposed to hypotonic media in the presence and absence of 5-nitro-2-(3-phenylpropylamino)benzoic acid or propofol. RESULTS: The relative fluorescence values at 10, 20, and 30% hypotonicity were 147 +/- 29, 214 +/- 74, and 335 +/- 102% of baseline, respectively. The changes were all significantly different from the isotonic time control group. In the presence of 200 microM 5-nitro-2-(3-phenylpropylamino)benzoic acid or 0.1, 1, 10, or 100 microg/ml propofol, the relative fluorescence values at 30% hypotonicity were 87 +/- 17, 194 +/- 27, 160 +/- 18, 130 +/- 18, and 84 +/- 15%, respectively. These changes were significantly less than the 30% for the hypotonic control (246 +/- 23%). CONCLUSION: These results suggest that volume-sensitive chloride channels and nonselective cation channels may participate in hypotonicity-induced membrane depolarization and that propofol inhibits hypotonicity-induced membrane depolarization in coronary artery smooth muscle. PMID- 15108982 TI - Single amino acid residue in the extracellular portion of transmembrane segment 2 in the nicotinic alpha7 acetylcholine receptor modulates sensitivity to ketamine. AB - BACKGROUND: Ketamine inhibits the activation of both heteromeric and homomeric nicotinic acetylcholine receptors. The site of molecular interaction is unknown. METHODS: The inhibition of alpha7 nicotinic acetylcholine receptors by ketamine was compared to that of 5-hydroxytryptamine-3A (5HT3A) receptors that are resistant to ketamine inhibition in Xenopus laevis oocytes. To determine whether the region of transmembrane segments 2 and 3 is relevant for ketamine inhibition of nicotinic receptors, the authors identified single amino acid residues that differ in the sequence alignment of the two proteins. They created 22 mutant alpha7 nicotinic receptors that contain the single homologous amino acid residue in the 5HT3A sequence. RESULTS: Of the 22 mutant alpha7 nicotinic receptors tested, only one (alpha7 A258S) was significantly resistant to 20 microM ketamine. The ketamine concentration response relationship for the alpha7 A258S mutant was shifted to the right with the IC50 for ketamine increased from 17 +/- 2 for wild type to 30 +/- 3 microM in the mutant (P < 0.001). Agonist activation was unchanged by the mutation. The homologous amino acid residue in the 5HT3A receptor was mutated to the alanine that occurs in the wild-type nicotinic receptor. This mutation made the previously insensitive 5HT3A receptor sensitive to ketamine (P < 0.001). CONCLUSIONS: Conservative mutation of a single amino acid in the extracellular transmembrane segment 2 domain induces resistance to ketamine inhibition in the alpha7 nicotinic receptor and sensitivity to inhibition in the 5HT3A receptor. This region may represent a ketamine binding site in the alpha7 nicotinic receptor, or it may be an important transduction site for ketamine action. PMID- 15108983 TI - Isoflurane inhibits transmitter release and the presynaptic action potential. AB - BACKGROUND: Isoflurane inhibits the excitatory postsynaptic current (EPSC) at many synapses. Accumulated evidence suggests the involvement of a presynaptic mechanism. However, the extent of the presynaptic contribution has not been quantitatively studied. Furthermore, the mechanism underlying the presynaptic contribution remains unclear. METHODS: To estimate the presynaptic contribution, the authors compared the effects of isoflurane on the presynaptic capacitance jump, which is proportional to vesicle release, and the postsynaptic glutamate receptor-mediated EPSC at a calyx-type synapse in rat brainstem. The authors determined whether isoflurane affects the waveform of the action potential recorded from nerve terminals. By studying the relation between the EPSC and the presynaptic action potential at the same synapse, the authors determined whether isoflurane inhibits the EPSC by decreasing the presynaptic action potential. RESULTS: Isoflurane at 0.35-1.05 mM reduced the EPSC and the presynaptic capacitance jump to a similar degree without affecting the miniature EPSC (an indicator of quantal size), suggesting that isoflurane inhibits the EPSC predominantly by reducing glutamate release. Isoflurane reduced the presynaptic action potential by approximately 3-8%. The EPSC was proportional to the presynaptic action potential amplitude raised to a power of 10.2. Based on this relation, inhibition of the presynaptic action potential contributed to 62-78% of isoflurane-induced inhibition of the EPSC. CONCLUSIONS: Isoflurane inhibits the EPSC predominantly by inhibition of transmitter release. Isoflurane reduces the presynaptic action potential amplitude, which may contribute significantly to its inhibitory effect on the EPSC. PMID- 15108984 TI - Peripheral nerve injury sensitizes the response to visceral distension but not its inhibition by the antidepressant milnacipran. AB - BACKGROUND: Manipulations that cause hypersensitivity to visceral stimuli have been shown to also result in hypersensitivity to somatic stimuli coming from convergent dermatomes, but the converse has not been examined. The authors tested whether lumbar spinal nerve ligation in rats, a common model of neuropathic pain that results in hypersensitivity to somatic stimuli, also leads to hypersensitivity to visceral stimuli coming from convergent dermatomes and whether pharmacology of inhibition differed between these two sensory modalities. METHODS: Female Sprague-Dawley rats were anesthetized, and the left L5 and L6 spinal nerves were ligated. Animals received either intrathecal saline or milnacipran (0.1-3 microg), and withdrawal thresholds to mechanical testing in the left hind paw, using von Frey filaments, and visceral testing, using balloon colorectal distension, were determined. RESULTS: Nerve ligation resulted in decreases in threshold to withdrawal to somatic mechanical stimulation (from 13 +/- 1.8 g to 2.7 +/- 0.7 g) and also in decreases in threshold to reflex response to visceral stimulation (from 60 mmHg to 40 mmHg). Intrathecal milnacipran increased withdrawal threshold to somatic stimulation in a dose-dependent manner but failed to alter the response to noxious visceral stimulation. CONCLUSIONS: Injury of nerves innervating somatic structures enhances nociception from stimulation of viscera with convergent input from nearby dermatomes, suggesting that somatic neuropathic pain could be accompanied by an increased likelihood of visceral pain. Lack of efficacy of the antidepressant milnacipran against visceral stimuli suggests that visceral hypersensitivity may not share the same pharmacology of inhibition as somatic hypersensitivity after nerve injury. PMID- 15108985 TI - ED50 and ED95 of intrathecal hyperbaric bupivacaine coadministered with opioids for cesarean delivery. AB - BACKGROUND: Successful cesarean delivery anesthesia has been reported with use of small doses (5-9 mg) of intrathecal bupivacaine coadministered with opioids. This double-blind, randomized, dose-ranging study determined the ED50 and ED95 of intrathecal bupivacaine (with adjuvant opioids) for cesarean delivery anesthesia. METHODS: Forty-two parturients undergoing elective cesarean delivery with use of combined spinal-epidural anesthesia received intrathecal hyperbaric bupivacaine in doses of 6, 7, 8, 9, 10, 11, or 12 mg in equal volumes with an added 10 microg intrathecal fentanyl and 200 microg intrathecal morphine. Sensory levels (pinprick) were evaluated every 2 min until a T6 level was achieved. The dose was a success(induction) if a bilateral T6 block occurred in 10 min; otherwise, it was a failure(induction). In addition to being a success(induction), the dose was a success(operation) if no intraoperative epidural supplement was required; otherwise, it was a failure(operation). ED50 and ED95 for both success(induction) and success(operation) were determined with use of a logistic regression model. RESULTS: ED50 for success(induction) and success(operation) were 6.7 and 7.6 mg, respectively, whereas the ED95 for success(induction) and success(operation) were 11.0 and 11.2 mg. Speed of onset correlated inversely with dose. Although no clear advantage for low doses could be demonstrated (hypotension, nausea, vomiting, pruritus, or maternal satisfaction), this study was underpowered to detect significance in these variables. CONCLUSIONS: The ED95 of intrathecal bupivacaine under the conditions of this study is considerably in excess of the low doses proposed for cesarean delivery in some recent publications. When doses of intrathecal bupivacaine less than the ED95, particularly near the ED50, are used, the doses should be administered as part of a catheter-based technique. PMID- 15108986 TI - Thoracic and lumbar epidural analgesia via the caudal approach using electrical stimulation guidance in pediatric patients: a review of 289 patients. AB - BACKGROUND: Nerve stimulation guidance (Tsui test) has been reported to be an effective alternative to radiographic imaging for proper catheter placement. The purpose of this study was to examine the success rate and complications of continuous caudal epidural analgesia since the implementation of routine use of the Tsui test at the authors' institution. METHODS: The authors examined prospectively collected data in their pediatric pain service database from 289 children who had attempted caudal placement of a lumbar or thoracic catheter between 1999 and 2002. RESULTS: In five patients (aged 5 months-1.6 yr), the catheter did not thread to the desired level and was abandoned in the operating room (technical success rate, 98.2%). Of the remaining 284 patients, the overall analgesic success rate of all caudal route epidural analgesia procedures was 84.9%. There was no significant difference in adequate pain control (success) in infants (aged 1 day-1 yr) versus older children (aged younger than 1 yr). The most common adverse effects were pruritus (26.1%) and nausea and vomiting (16.9%). Of the patients in our study, 57.7% had urinary catheters in situ; of those who did not have a catheter placed, 20.8% experienced urinary retention. The incidence of respiratory depression was 4.2%, but the administration of naloxone for severe respiratory depression was never necessary. Three percent of catheters were removed because of suspected contamination, but no epidural abscesses or systemic infection were noted. CONCLUSIONS: The results of this study suggest that epidural catheter placement via the caudal approach using the Tsui test is an effective and reasonable alternative to direct lumbar and thoracic epidural analgesia in pediatric patients. PMID- 15108987 TI - Synergistic antinociceptive effect of amitriptyline and morphine in the rat orofacial formalin test. AB - BACKGROUND: Combination therapy is often used to increase the clinical utility of analgesic agents. The coadministration of two compounds may achieve analgesia at doses lower than those required for either compound alone, leading to enhanced pain relief and reduction of adverse effects. Herein, the authors describe the effect of coadministration of morphine and amitriptyline on cutaneous orofacial inflammatory pain in rats. METHODS: Amitriptyline, morphine, or the combination of amitriptyline and morphine was administered systemically to rats, and antinociceptive effects were determined by means of the rat orofacial formalin test. Isobolographic analysis was used to define the nature of the interactions between morphine and amitriptyline. RESULTS: Amitriptyline as well as morphine produced a dose-related inhibition in the first phase and the second phase of rubbing activity. ED50 values against rubbing behavior were 14.6 mg/kg (95% confidence interval, 10.2-33.5 mg/kg) and 1.3 mg/kg (95% confidence interval, 1.0 1.7 mg/kg) for amitriptyline and morphine, respectively. Combinations of increasing fractional increments of amitriptyline and morphine ED50 doses produced a synergistic effect against rubbing behavior, as revealed by isobolographic analysis. CONCLUSIONS: The current study suggests that systemic amitriptyline and morphine synergistically inhibit cutaneous orofacial inflammatory pain in rats. PMID- 15108988 TI - Economics of nerve block pain management after anterior cruciate ligament reconstruction: potential hospital cost savings via associated postanesthesia care unit bypass and same-day discharge. AB - BACKGROUND: Anterior cruciate ligament reconstruction is a complex outpatient surgical procedure often associated with pain. Traditionally, the procedure is performed under general anesthesia and often requires the use of the PACU. Refractory pain and/or nausea/vomiting occasionally leads to an unplanned hospital admission. In this study, the authors examine the associations of nerve block analgesia for these patients and its associated reductions in PACU use, hospital admission, and hospital costs. METHODS: This was an observational, nonrandomized study in which existing data regarding patients' day-of-surgery outcomes were merged with hospital cost data. We reviewed a consecutive sample of 948 men and women who were in good health and underwent anterior cruciate ligament reconstruction in an outpatient surgery unit between July 1995 and June 1999. RESULTS: The use of nerve block analgesia was associated with reduced PACU admissions to 18% and decreased unplanned hospital admission rates from 17% to 4%. Multivariate linear regression analysis showed that patients bypassing the PACU had an associated hospital cost reduction of 12% (P = 0.0001), whereas patients who needed hospital admission had an associated hospital cost increase of 11% (P = 0.0003). CONCLUSIONS: The use of nerve blocks for acute pain management in patients undergoing anterior cruciate ligament reconstruction is associated with PACU bypass and reliable same-day discharge. Although the cost savings for this one procedure are unlikely to generate sufficient cost savings via staffing reductions, extrapolating these results to a large volume of all types of invasive outpatient orthopedic procedures may have the potential to create significant hospital cost savings. PMID- 15108989 TI - Mechanisms of cardioprotection by volatile anesthetics. PMID- 15108990 TI - Current concepts of hemostasis: implications for therapy. AB - The revised model of coagulation has implications for therapy of both hemorrhagic and thrombotic disorders. Of particular interest to anesthesiologists is the management of clotting abnormalities before, during, and after surgery. Most hereditary and acquired coagulation factor deficiencies can be managed by specific replacement therapy using clotting factor concentrates. Specific guidelines have also been developed for perioperative management of patients using anticoagulant agents that inhibit platelet or coagulation factor functions. Finally, recombinant factor VIIa has been used off-label as a hemostatic agent in some surgical situations associated with excessive bleeding that is not responsive to conventional therapy. PMID- 15108991 TI - Spontaneous occurrence of the disposition to malignant hyperthermia. PMID- 15108992 TI - A bifurcated tracheal tube for a neonate with tracheoesophageal fistula. PMID- 15108993 TI - Anesthetizing the phantom: peripheral nerve stimulation of a nonexistent extremity. PMID- 15108994 TI - Dexmedetomidine and cardiac arrest. PMID- 15108995 TI - Electrical field stimulation to study inhibitory mechanisms in individual sensory neurons in culture. PMID- 15108996 TI - The pain visual analog scale: linear or nonlinear? PMID- 15108997 TI - Analgesic evaluation in postoperative patients. PMID- 15108998 TI - Is overestimation of bispectral index in sedated intensive care unit patients only related to electromyographic activity? PMID- 15108999 TI - Management of patient body temperature is challenging. PMID- 15109000 TI - The perioperative use of cyclooxygenase-2 selective nonsteroidal antiinflammatory drugs may offer a safer alternative. PMID- 15109001 TI - Effects of postoperative nonsteroidal antiinflammatory drugs on bleeding risk after tonsillectomy. PMID- 15109002 TI - Nonsteroidal antiinflammatory drugs and hemorrhage following tonsillectomy: do we have the data? PMID- 15109003 TI - Carotid sinus mechanical properties. PMID- 15109004 TI - Unusual case of breathing circuit obstruction: plastic packaging revisited. PMID- 15109005 TI - Revenue gain for academic anesthesiology departments if the Centers for Medicare and Medicaid Services provide full reimbursement to teaching physicians. PMID- 15109006 TI - Deviation of the cauda equina by changing position. PMID- 15109007 TI - Novel breathing circuit architecture: new consequences of old problems. PMID- 15109008 TI - Magnetic resonance spectroscopy and clinical cancer prognosis. PMID- 15109009 TI - Predicting treatment response in non-Hodgkin's lymphoma from the pretreatment tumor content of phosphoethanolamine plus phosphocholine. AB - RATIONALE AND OBJECTIVES: Phosphoethanolamine and phosphocholine, shown to be elevated in tumors and possibly related to apoptotic signaling, have the potential to be prognostic variables of cancer treatment. MATERIALS AND METHODS: The sum of phosphoethanolamine and phosphocholine normalized by nucleotide triphosphates was determined in tumors of non-Hodgkin's lymphoma (NHL) patients via in vivo 31P MR spectroscopy. RESULTS: The normalized sum of phosphoethanolamine and phosphocholine showed significant differences in tumors of patients who had a complete response to treatment against those who did not (t test: 1.45 +/- 0.15, mean +/- standard error, n = 10 vs. 2.28 +/- 0.15, n = 17, P < .001; Fisher test: P < .04; sensitivity and specificity approximately equal to 70%). This parameter also showed significant differences among treatment responses in the previously untreated and aggressive subgroups and in the low and low-intermediate-risk subgroups determined by the international prognostic index (IPI). Further, distinctly different treatment response cutoffs for the parameter were found in different risk groups. When these risk-dependent cutoffs were used, the Fisher test of the whole group improved (P < .0002, sensitivity 80%, specificity 94%). The normalized sum of phosphoethanolamine and phosphocholine and the IPI were better predictor covariates for time to treatment failure when fitted interactively in a Cox regression (P < .0003) than when fitted independently. When time to treatment failure was used as a surrogate of survival in Kaplan-Meier analysis, the interaction of both covariates segregated the cases significantly (P < .008). There was no significance with each covariate independently. CONCLUSION: The normalized sum of phosphoethanolamine and phosphocholine measured before treatment successfully predicts long-term response to treatment and time to treatment failure in non-Hodgkin's lymphoma, particularly when combined with the IPI. PMID- 15109010 TI - Area beam equalization: optimization and performance of an automated prototype system for chest radiography. AB - RATIONALE AND OBJECTIVES: To evaluate the feasibility and performance of an x-ray beam equalization system for chest radiography using anthropomorphic phantoms. MATERIALS AND METHODS: Area beam equalization involves the process of the initial unequalized image acquisition, attenuator thickness calculation, mask generation using a 16 x 16 piston array, and final equalized image acquisition. Chest radiographs of three different anthropomorphic phantoms were acquired with no beam equalization and equalization levels of 4.8, 11.3, and 21. Six radiologists evaluated the images by scoring them from 1-5 using 13 different criteria. The dose was calculated using the known attenuator material thickness and the mAs of the x-ray tube. RESULTS: The visibility of anatomic structures in the under penetrated regions of the chest radiographs was shown to be significantly (P < .01) improved after beam equalization. An equalization level of 4.8 provided most of the improvements with moderate increases in patient dose and tube loading. Higher levels of beam equalization did not show much improvement in the visibility of anatomic structures in the under-penetrated regions. CONCLUSION: A moderate level of x-ray beam equalization in chest radiography is superior to both conventional radiographs and radiographs with high levels of beam equalization. X-ray beam equalization can significantly improve the visibility of anatomic structures in the under-penetrated regions while maintaining good image quality in the lung region. PMID- 15109011 TI - Effect of sex and gender on drug-seeking behavior during invasive medical procedures. AB - RATIONALE AND OBJECTIVES: To assess how sex affects patients' drug-seeking, pain, and anxiety during interventional procedures. MATERIALS AND METHODS: Data from 159 patients were derived from two control groups of a prospective randomized trial. Seventy-six patients were male, 83 female. Patients in the standard group (n = 79) received the standard care typical for the institution; patients in the attention group (n = 80) had an additional empathic provider who stayed with them throughout the procedure. All patients were asked every 15 minutes to rate their pain and anxiety on 0-10 self-rating scales. All had access to intravenous sedatives and narcotics through a patient-controlled analgesia model. Univariate analysis of variance with a between-patient factor for group and another between patient factor for sex was used. RESULTS: There was a significant interaction between group attribution and sex with regard to drug request and pain and anxiety ratings. Patients in the attention group requested significantly fewer drugs than patients in the standard group. Men asked for more drugs than women under standard care, but for less in the attention group. Pain and anxiety ratings for women were significantly lower in the attention group compared with standard treatment, but for men, there was no significant difference. CONCLUSION: Although both men and women benefit from the presence of an empathic provider during invasive medical procedures, men benefit more in terms of medication reduction, whereas women benefit more in terms of pain and anxiety reduction. Awareness of these gender-specific differences can aid in formulation of patient specific treatment plans. PMID- 15109012 TI - Detection and classification performance levels of mammographic masses under different computer-aided detection cueing environments. AB - RATIONALE AND OBJECTIVES: The authors evaluated the impact of different computer aided detection (CAD) cueing conditions on radiologists' performance levels in detecting and classifying masses depicted on mammograms. MATERIALS AND METHODS: In an observer performance study, eight radiologists interpreted 110 subtle cases six times under different display conditions to detect depicted masses and classify them as benign or malignant. Forty-five cases depicted biopsy-proven masses and 65 were negative. One mass-based cueing sensitivity of 80% and two false-positive cueing rates of 1.2 and 0.5 per image were used in this study. In one mode, radiologists first interpreted images without CAD results, followed by the display of cues and reinterpretation. In another mode, radiologists viewed CAD cues as images were presented and then interpreted images. Free-response receiver operating characteristic method was used to analyze and compare detection performance. The receiver operating characteristic method was used to evaluate classification performance. RESULTS: At these performance levels, providing cues after initial interpretation had little effect on the overall performance in detecting masses. However, in the mode with the highest false positive cueing rate, viewing CAD cues immediately upon display of images significantly reduced average performance for both detection and classification tasks (P < .05). Viewing CAD cues during the initial display consistently resulted in fewer abnormalities being identified in noncued regions. CONCLUSION: CAD systems with low sensitivity (< or = 80% on mass-based detection) and high false-positive rate (> or = 0.5 per image) in a dataset with subtle abnormalities had little effect on radiologists' performance in the detection and classification of mammographic masses. PMID- 15109013 TI - Experiences with a workstation prototype for softcopy reading within the Bavarian mammography recertification program: workstations and education. AB - RATIONALE AND OBJECTIVES: In January 2002, the Bavarian Statutory Health Care Administration ("Kassenarztliche Vereinigung Bayerns", KVB) started a recertification program for quality assurance and quality improvement in mammography reading. MATERIALS AND METHODS: All accredited radiologists and gynecologists are asked to prove their qualification every 1-2 years. The recertification program requires the physicians to read 50 cases randomly selected from a larger collection of high-quality test cases. The portion of malignant and benign cases corresponds to the requirements of the German National Association of Statutory Health Insurance Physicians ("Kassenaztliche Bundesvereinig ung", KBV). In order to perform the recertification in a manageable manner a decentralized approach (test at different locations in parallel) was preferred over a centralized one. Therefore, the X-ray films were digitized and converted to DICOM Digital Mammography format to be read on a softcopy device. To verify the applicability of digitized mammograms for recertification purposes, a comparative study with 32 trained radiologists and gynecologists was performed. RESULTS: A system of two high-resolution/high contrast monitors (2048 X 2560 pixels, > or = 350 cd/m2) in combination with a 5 mega-pixel dual-head graphics adapter with calibrated output was chosen for the mammography workstation. The software was implemented according to the particular requirements of this program. As a result, the comparative study showed that there was no significant difference in the error rate of the reported findings between conventional film and softcopy reading. CONCLUSION: The first intermediate results of this quality initiative are promising. As of 2003, the test is mandatory for all mammography-reading physicians in Bavaria. PMID- 15109014 TI - 3D volume segmentation of MRA data sets using level sets: image processing and display. AB - In this article, we use a level set-based segmentation algorithm to extract the vascular tree from magnetic resonance angiography (MRA) data sets. The classification approach depends on initializing the level sets in the 3D volume, and the level sets evolve with time to yield the blood vessels. This work introduces a high-quality initialization for the level set functions, allowing extraction of the blood vessels in 3D and elimination of non-vessel tissues. A comparison between the 2D and 3D segmentation approaches is made. The results are validated using a phantom that simulates the MRA data and show good accuracy. PMID- 15109015 TI - A survey evaluating the beliefs of radiology residency program directors regarding incorporation of numeric standards into accreditation standards. AB - RATIONALE AND OBJECTIVES: Numeric standards do not exist for the evaluation and comparison of radiology resident education. The Accreditation Council for Graduate Medical Education has recently proposed the evaluation and assessment of residents by using numeric values for films read in various radiology modalities. The purpose of this study was to evaluate radiology residency program directors' perceived need for numeric standards for radiology residency programs. MATERIALS AND METHODS: Program directors were surveyed via a three-part questionnaire. Initial contact was made by e-mail, with follow-up by telephone if necessary. The survey questions were: (1) Do you feel that a need exists for a set of numerical standards for the number of films read by radiology residents in each of the various subspecialties? (2) Specify the needs that you see. (3) What are the barriers and disadvantages to acquiring such data? RESULTS: Survey responses were received from 44 of 192 radiology residency programs (23% response rate). Overall, 34 (77%) of 44 program directors were against implementation of the Accreditation Council for Graduate Medical Education proposal, whereas 8 (18%) of 44 program directors were in favor of the proposal and 2 (5%) of 44 were undecided. A variety of reasons, both for and against the proposal, were provided by program directors. CONCLUSION: Among the program directors responding to the survey, there was strong opposition to implementation of mandatory standards for specific numbers of radiologic examinations by residents as the basis for program accreditation. PMID- 15109016 TI - Implementing a full-feature PACS solution in accordance with the IHE technical framework: the CHILI approach. AB - RATIONALE AND OBJECTIVES: During the last 4 years, Integrating the Healthcare Enterprise (IHE) has worked in establishing a reliable integrated environment for medical imaging devices and other clinical information systems by using existing industry standards such as Digital Imaging and Communication in Medicine (DICOM) and Health Level Seven (HL7). Because IHE is organized and driven by professional organizations representing both buyers and vendors, it was able to define a common language for the various health care parties who are involved in the integration process. Thus IHE offers a rigorously organized technical framework that provides a comprehensive guide for a coordinated implementation of information standards. MATERIALS AND METHODS: A multistage plan for incorporating the IHE elements while scaling up general-purpose workstations with teleradiology functionalities to a full-feature picture archiving and communication system (PACS) solution was created. To introduce a pragmatic example, the plan approach was applied to the CHILI workstations (CHILI, Heidelberg, Germany). To implement the proposed plan in making various legacy systems comply with IHE, the effects of available resources and market needs on the plan are discussed. Most implementation challenges and problems were overcome using generic design approaches and well-designed DICOM utilities. RESULTS: On completion of the first stage, the PACS viewer and reporting workstations were IHE-compliant. In addition, the plan facilitates the conformance process while maintaining the planned software development cycle. CONCLUSION: Based on these positive results and the practical experience acquired during the first stage, further stages will be completed to provide the system with the required plug-and-play interoperability among systems from different vendors. PMID- 15109017 TI - A primer on molecular biology for imagers: III. Proteins: structure and function. AB - This article along with the first 2 in this series (4,12) completes the discussion on the key molecules and process inside the cell namely, DNA, RNA, and proteins. These 3 articles provide a very basic foundation for understanding molecular biology concepts and summarize some of the work of numerous scientists over the past century. We understand these processes far better now than we did in the past, but clearly this knowledge is by no means complete and a number of basic scientists are working hard to elucidate and understand the fundamental mechanisms that operate within a cell. Genes and gene products work with each other in complex, interconnected pathways, and in perfect harmony to make a functional cell, tissue, and an organism as a whole. There is a lot of cross-talk that happens between different proteins that interact with various other proteins, DNA, and RNA to establish pathways, networks, and molecular systems as a team working to perfection. The past 15 years have seen the rapid development of systems biology approaches. We live in an era that emphasizes multi disciplinary, cross-functional teams to perform science rather than individual researchers working on the bench on a very specific problem. Global approaches have become more common and the amount of data generated must be managed by trained bioinformatics personnel and large computers. In our subsequent articles, we will discuss these global approaches and the areas of genomics, functional genomics, and proteomics that have revolutionized the way we perform science. PMID- 15109018 TI - Assessment methodologies and statistical issues for computer-aided diagnosis of lung nodules in computed tomography: contemporary research topics relevant to the lung image database consortium. AB - Cancer of the lung and bronchus is the leading fatal malignancy in the United States. Five-year survival is low, but treatment of early stage disease considerably improves chances of survival. Advances in multidetector-row computed tomography technology provide detection of smaller lung nodules and offer a potentially effective screening tool. The large number of images per exam, however, requires considerable radiologist time for interpretation and is an impediment to clinical throughput. Thus, computer-aided diagnosis (CAD) methods are needed to assist radiologists with their decision making. To promote the development of CAD methods, the National Cancer Institute formed the Lung Image Database Consortium (LIDC). The LIDC is charged with developing the consensus and standards necessary to create an image database of multidetector-row computed tomography lung images as a resource for CAD researchers. To develop such a prospective database, its potential uses must be anticipated. The ultimate applications will influence the information that must be included along with the images, the relevant measures of algorithm performance, and the number of required images. In this article we outline assessment methodologies and statistical issues as they relate to several potential uses of the LIDC database. We review methods for performance assessment and discuss issues of defining "truth" as well as the complications that arise when truth information is not available. We also discuss issues about sizing and populating a database. PMID- 15109019 TI - The role of technology in radiology education. PMID- 15109020 TI - Radiology is a perfect tool for problem based learning. PMID- 15109021 TI - X equals something. PMID- 15109022 TI - The green cigar. PMID- 15109023 TI - Nanoparticle engineering processes for enhancing the dissolution rates of poorly water soluble drugs. AB - Poor water solubility is an industry wide issue, especially for pharmaceutical scientists in drug discovery and drug development. In recent years, nanoparticle engineering processes have become promising approaches for the enhancement of dissolution rates of poorly water soluble drugs. Nanoparticle engineering enables manufacturing of poorly water soluble drugs into nanoparticles alone, or incorporation with a combination of pharmaceutical excipients. The use of these processes has dramatically improved in vitro dissolution rates and in vivo bioavailabilities of many poorly water soluble drugs. This review highlights several commercially or potentially commercially available nanoparticle engineering processes recently reported in the literature for increasing the dissolution properties of poorly water soluble drugs. PMID- 15109024 TI - Safety and efficacy: the role of chemistry, manufacturing, and controls in pharmaceutical drug development. PMID- 15109025 TI - Comparison of low shear, high shear, and fluid bed granulation during low dose tablet process development. AB - Three processing methods were compared to develop a low dose (0.1%) immediate release tablet. Similar formulations were used to evaluate low shear, high shear, and fluid bed granulation methods. For each granulation process, the drug was dissolved or suspended in the granulating fluid and sprayed into the granulator. Both water and methanol were evaluated as granulating fluids. The low shear granulation was performed in a Patterson-Kelley V-Blender with I-bar. The high shear granulation was performed in a GRAL (top entry impeller) and a Diosna (bottom mounted impeller). Fluid bed granulation was also performed using top spray. Acceptable content uniformity was obtained using each technology. The type of granulator and granulating solvent affected the granulation particle size distributions and bulk/tap densities. However, the addition of extragranular microcrystalline cellulose minimized the effect of variable granulation properties and allowed similar tablets to be produced from each granulation process. PMID- 15109026 TI - Influence of the valve lubricant on the aerodynamic particle size of a metered dose inhaler. AB - Presented in this work are the results of a study designed to investigate the impact of valve lubricant (i.e., silicone oil) on the aerodynamic particle size distribution (PSD) of a steroid suspension metered dose inhaler (MDI) containing propellant HFA-227. The objective of this study was to explore whether the valve lubricant, which is often used in MDI products to prevent valve sticking, can enter an MDI product and potentially impact the aerosol spray dynamics. The results of this work have shown that samples containing valves with high silicone levels produced a larger aerodynamic particle size (by cascade impaction) than samples with low-silicone or silicone-free valves. It is postulated that the presence of silicone in the product may increase the propensity for drug aggregation, thereby leading to an increase in the aerodynamic particle size of the emitted aerosol. These findings stress the importance of evaluating the effects of valve lubricant on the aerodynamic PSD in the early formulation development stage of an MDI. PMID- 15109027 TI - Characterization of the mechanism of interaction in ibuprofen-Eudragit RL100 coevaporates. AB - The present study is a preliminary exploration of the affinity between a carboxylic model drug, the nonsteroidal antiinflammatory agent ibuprofen (IBU) and Eudragit RL100 (RL) polymer. Due to the presence of a variable amount of quaternary ammonium groups in this matrix, physical and chemical interaction with the carboxylic drug can occur, which reinforces its scant mechanical dispersion in the polymer network and can ultimately affect its release profile in vitro and in vivo. To study these aspects, IBU was mixed at increasing weight ratios and in different chemical forms (free acid, sodium salt, and n-butyl ester), to investigate further the role of the carboxylic group in the interaction with the RL polymer. Therefore, IBU-RL solid dispersions (coevaporates) were obtained and fully characterized in the solid state through spectroscopic, calorimetric, and x ray diffractometric analyses. The in vitro release pattern of the drug, in the different chemical states, was studied for the coevaporates, compared with drug RL physical mixtures, along with drug adsorption profiles from aqueous solutions on the surface of the polymer granules. PMID- 15109028 TI - In vitro skin permeation and retention of paromomycin from liposomes for topical treatment of the cutaneous leishmaniasis. AB - Paromomycin (PA), a very hydrophilic antibiotic, has been tested as an alternative topical treatment against cutaneous leishmaniasis (CL). Although this treatment has shown promising results, it has not been successful in accelerating the recovery in most cases. This could be attributed to the low skin penetration of PA. Liposomal formulations usually provide sustained and enhanced drug levels in skin. The aim of this study was to prepare liposomal formulations containing PA and to investigate their potential as topical delivery systems of this antileishmanial. Large multilamellar vesicles (MLVs) were prepared by conventional solvent evaporation method. Large unilamellar vesicles (LUVs) were prepared by reverse-phase evaporation method. The lipids used were soybean phosphatidylcholine (PC) and PC:cholesterol (CH) (molar ratio 1:1). The skin permeation experiments across stripped and normal hairless mice skin were performed in modified Franz diffusion cells. The PA entrapment in LUV liposomes (20.4 +/- 2.2%) was higher than that observed for MLV liposomes (7.5 +/- 0.9%). Drug entrapment was 41.9 +/- 6.2% and 27.2 +/- 2.4% for PC and PC:CH LUV, respectively. The skin permeation was 1.55 +/- 0.31%, 1.29 +/- 0.40%, 0.20 +/- 0.08%, and 0.50 +/- 0.19% for PC LUV, PC:CH LUV, empty LUV +/- PA and aqueous solution, respectively. Controlled topical delivery, across stripped skin, was observed for PA entrapped in LUV liposomes. PMID- 15109029 TI - Effect of hydroxypropyl beta-cyclodextrin on drug solubility in water-propylene glycol mixtures. AB - Combined effects of cosolvency and inclusion complexation on drug solubility were studied using a model hydrophobic compound (carbamazepine) and a model hydrophilic compound (Compound S). Propylene glycol (PG) was used as the nonaqueous solvent, and deionized water was employed for the aqueous systems. Hydroxypropyl beta-cyclodextrin (HPbetaCD) was chosen as the complexing agent and studied at concentrations up to 28% (w/v). Complex formation constants (Kc) and solubility enhancement ratios were determined for the respective compounds in various water/PG vehicles. The data suggested that the inclusion of the compounds was most favorable when water alone was used as the vehicle. However, the combined approach of cosolvency and complexation resulted in a significant increase in the total apparent solubility of carbamazepine (the hydrophobic compound). The same was not observed with Compound S (the hydrophilic model), since PG weakened the interactions between the molecule and HPbetaCD, and thus, no synergistic or additive effects were observed with the combined approach of complexation and cosolvency. PMID- 15109030 TI - Physicochemical characterization of solid dispersions of indomethacin with PEG 6000, Myrj 52, lactose, sorbitol, dextrin, and Eudragit E100. AB - The purpose of this study was to prepare and characterize solid dispersions of indomethacin with polyethylene glycol (PEG) 6000, Myrj 52, Eudragit E100, and different carbohydrates such as lactose, mannitol, sorbitol, and dextrin. Indomethacin is a class II substance according to the Biopharmaceutics Classification System. It is a poorly water soluble antirheumatic agent. The goal was to investigate whether the solid dispersion can improve the dissolution properties of indomethacin. The solid dispersions were prepared by three different methods depending on the type of carrier. The evaluation of the properties of the dispersions was performed using solubility measurements, dissolution studies, Fourier-transform infrared spectroscopy, and x-ray powder diffractometery. The results indicate that lactose, mannitol, sorbitol, and especially Myrj 52 are suitable carriers to enhance the in vitro dissolution rate of indomethacin at pH 7.2. Eudragit E100, Myrj 52, and mannitol increase the dissolution properties at pH 1.2. The data from the x-ray diffraction showed that the drug was still detectable in its solid state in all solid dispersions except solid dispersions with dextrin and high amounts of mannitol. However, the results from infrared spectroscopy together with those from x-ray diffraction showed well defined drug-carrier interactions for dextrin coevaporates. PMID- 15109031 TI - Design and atomization properties for an inside-out type effervescent atomizer. AB - Atomization of aqueous polymer solutions is a key step in the formulation of several pharmaceutical products. Droplet size control is essential in order to produce pharmaceutical products with the desired properties. The purpose of this paper is to investigate design issues for an inside-out type of effervescent atomizer used to spray water and aqueous solutions of polyvinylpyrrolidone (Kollidon K-30) and hydroxypropyl methylcellulose (Pharmacoat 603). The atomizer was operated at air-to-liquid mass ratios of 0.1, 0.3, and 0.5 and a feed pressure of 1172 kPa. Fluid viscosities ranged from 1 to 47 mPa.s. The influence of several atomizer design features was considered, including exit orifice length to-diameter ratio, exit orifice diameter, the total area of the air injection holes, the distance between the air injection point and the exit orifice, the diameter of the mixing chamber, and the orientation of both air and liquid flows. Droplet size distributions were shown to vary significantly with the atomizer's exit orifice diameter, air injector design, and air injector distance to the exit orifice. In all cases air-to-liquid mass ratio played a key role in the mean droplet size. The design of the atomizer was shown to have the most pronounced effect on the mean droplet size at the lowest air-to-liquid mass ratios. Optimization of the atomizer design is very important in order to obtain small droplet sizes in pharmaceutical processes where the amount of air/gas should be minimized, e.g., closed-cycle spray drying and agglomeration processes. PMID- 15109032 TI - [Buried Pleistocene human remains of Weissenthurm, Mayen-Koblenz district: new and old information]]. AB - The landscape at central Rhine and Mosel is one of the most famous archaeological sites in middle Europe. A layer of pumicetufa from the eruption of the lake Laacher volcano 13,000 years B.P. is an important mark which approximately divides the Palaeolithic and Mesolithic period. Although numerous excavations in this area have been carried out, quaternary hominid remains are quite rare. A few short notes from the early 1920s reports of human bones "below the pumice, in Weissenthurm, District Mayen-Koblenz, Rhineland-Palatinate". However, these remains were probably destroyed in the Second World War in Munich on April 25, 1944. Recently, some new information has appeared on the discovery and the whereabouts of these fragments. The chronological classification of the Weissenthurm-hominid into the Pleistocene based on this information remains uncertain. PMID- 15109033 TI - Review of horses in Paleoindian sites of the Americas. AB - Analysis of the morphological characters in North and South American horses present during Paleoindian time indicates that at least eight Equus ecospecies occurred in North America. In South America, Equus had radiated into four ecospecies, Hippidion had one, and Onohippidium had three geographically separate ecospecies. These species are found in archeological deposits ranging from ca. 13,000 to 8,000 yr B.P., in tropical habitats as well as in the high Andean and Patagonian colder ecotopes. PMID- 15109034 TI - [Teenage pregnancies from the viewpoint of evolutionary biology]. AB - Teenage pregnancies have been considered mainly as perinatological and social problem up to now, in contrast biological and evolutionary aspects were neglected. In the present analysis the hypothesis is discussed that teenage pregnancies may represent a fitness maximizing strategy, with other words an efficient use of the relatively short female reproductive span. Results from auxology, primatology, historical demography and life history research, however, are in contradiction to the hypothesis. Extremely early teenage pregnancies (below the 16th birthday) seem to be a recent phenomenon caused by the decline of sexual maturation in course of the well described secular acceleration trend. PMID- 15109035 TI - Contribution of genetic and environmental factors to variation in body compartments--a twin study in adults. AB - This study aimed at analyzing the contribution of genetic and environmental factors on phenotypic variation of various traits of body composition. Subjects were 30 same-sexed pairs of twins including 20 monozygous (MZ) and 10 dizygous (DZ) pairs, aged 19-62 years. Zygosity was determined by DNA typing and morphological diagnosis. Body composition parameters (fat mass FM, lean body mass LBM, body cell mass BCM, extracellular mass ECM, total body water TBW, extracellular water ECW, and intracellular water ICW) were estimated by tetrapolar bioelectrical impedance analysis. Potential environmental factors influencing body composition (number of children, sporting activity and smoking behaviour) were determined by questionnaires. Heritabilities for traits of body composition were calculated by use of the twin method. Intraclass correlation is > 0.80 for the variation of LBM, BCM, ECM, TBW, ECW, and ICW in both MZ and DZ twins. Estimated heritability (h2) for FM, LBM, BCM, ECW, TBW, ECW, and ICW is 65%, 77%, 79%, 83%, 76%, 68%, and 82%, respectively. The h2 values for FM and LBM are consistent with those reported in other twin studies. For BCM, ECM, ECW and ICW, no comparative h2 estimates exist. Within-pair differences in body compartments do not change with increasing age in MZ and DZ twin pairs (p > 0.05). Stepwise multiple regression analyses indicate that zygosity, age, sex, number of children, sporting level and smoking behaviour do not significantly predict within-pair differences for weight, BMI, FM, LBM, TBW, ECW and ICW (each, p > 0.05). In contrast, sex and the number of children explain together 27% of observed within-pair differences for BCM. Zygosity is the only significant predictor of within-pair differences for ECM and height, explaining 20% (p = 0.008) and 36% of variance, respectively (p < 0.0001). Results indicate that genetic factors exert stronger influences on body composition than the considered environmental traits. PMID- 15109036 TI - Israeli jewish infants of different descent: growth patterns, likeness and differences. Longitudinal study. AB - BACKGROUND: Due to increasing migration process and intermarriages among individuals belonging to different ethnic groups, it is important to examine whether intermixing of populations effects child growth rate. AIMS: To compare growth patterns of Jewish infants from distinct descent. SUBJECTS: 1300 Israeli children were divided into 7 groups: 4 groups of babies with both parents from the same geographic origin (Europe, Yemen, Middle East or North Africa) and 3 inter-mixed groups according to genetic distances between the parents' derivation. STUDY DESIGN: The studied infants were monitored longitudinally for both body length. weight and head circumference from birth until 20 months of life. Using the curve fitting technique the follow-up data were fitted to the 3 parameter Count model. OUTCOME MEASUREMENTS: Three sets of maximal likelihood estimates of the model parameters were obtained to test the growth patterns of different groups of Jewish children: individually-specific for every child, group specific for each individual within the group, and general for all individuals from all studied groups. Likelihood ratio test was used to examine whether the chosen function of trait dependence on age is uniformly reliable for all individuals from all the considered groups. RESULTS AND CONCLUSIONS: Significant differences were indicated between all studied growth curves for all three studied traits. Moreover, distances calculated between the studied cohorts demonstrated a clear distinction between the clusters of "non-mixed" and "mixed" groups for length and weight. Since the studied children were born and developed in similar environments, these results may serve as important evidence for the existence of a genetic effect on the growth process. PMID- 15109037 TI - Reproducibility of anthropometric measurements in children: a longitudinal study. AB - The purpose of this study was to establish the reproducibility of a series of anthropometric measures performed twice during one week during a three year period in boys and girls. The subjects of this investigation were 39 children (21 boys and 18 girls), 9-10 year of age at the beginning of the study. Children were measured three times with one year interval. Children were classified by Tanner stage 1-2 during the first measurements, stage 1-3 during the second measurements and stage 1-4 during the third measurements. Body height and weight were measured and BMI calculated. All anthropometric parameters were measured according to the protocol recommended by the International Society for the Advancement of Kinanthropometry (Norton & Olds 1996). Nine skinfolds, 13 girths, eight lengths and eight breadths/lengths were measured. The reproducibility of body height (r = 0.995-0.999), body weight (r = 0.990-0.999) and BMI (r = 0.969-0.999) was very high in boys and girls. The intraclass correlations (ICC), technical errors (TE) and coefficients of variation (CV) were quite different depending on the measurement site of the skinfold thickness. It was surprising that the ICCs were highest and TEs and CVs were lowest during the second year of the measurement. The computed ICC was high, and TE and CV values were quite similar and relatively low in girth, length and breadth/length measurements. It was concluded that the reproducibility of girths, lengths and breadths/lengths in children is very high and the reproducibility of skinfolds is high. Specifically, the reproducibility is very high immediately before puberty in boys and girls. PMID- 15109038 TI - Body build classes as a method for systematization of age-related anthropometric changes in girls aged 7-8 and 17-18 years. AB - A total of 462 schoolgirls aged 7-8 and 17-18 years were examined anthropometrically (45 body measurements and 10 skinfolds) in a cross-sectional study. The data were processed in two age groups: 7-8-year-olds (n = 205) and 17 18-year-olds (n = 257). Relying on average height and weight in the groups, both groups were divided into five body build classes: small, medium, large, pyknomorphous and leptomorphous. In these classes, the differences in all other body measurements were compared, and in both age groups, analogous systematic differences were found in length, width and depth measurements and circumferences. This enabled us to compare proportional changes in body measurements during ten years, using for this ratios of averages of basic measurements and measurement groups in the same body build classes. Statistical analysis by the sign test revealed statistically significant differences between various body build classes in the growth of averages. Girls belonging to the small class differed from the girls of the large class by an essentially greater increase in their measurements. Our results suggest that the growth rate of body measurements of girls with different body build can be studied by the help of body build classification. PMID- 15109039 TI - Estimation of human endurance time. AB - A prediction formula has been evolved for estimation of human endurance time from aerobic and anaerobic fraction of the total oxygen utilization. The derivation of the formula is based on the assumption that fractional change in endurance time varies directly as the fractional change in aerobic fraction in the same direction and varies as the fractional change in anaerobic fraction in the opposite direction. The validity of the prediction formula has been tested on two sets of data. The first set is consisting of 31 observations on 13 Indian subjects and a second set of data is consisting of 7 observations on one subject collected from literature. The multiple correlations for these sets of data were 0.9650 and 0.9996, respectively. These multiple correlations were highly significant (p < 0.001). It has been concluded that aerobic and anaerobic fractions of total oxygen utilization are significant predictors of human endurance time. PMID- 15109040 TI - Promoting oral health in elder care--challenges and opportunities. PMID- 15109041 TI - Looking past and looking forward to the preferred gerontologic nursing future- 2003 Doris Schwartz Award presentation. PMID- 15109042 TI - Diabetic foot care in a long-term facility. AB - Foot problems continue to be a major cause of morbidity, disability, and mortality for individuals with diabetes. According to Rothman's model of causation, as interpreted by Pecoraro, Reiber, and Burges (1990), each amputation related to diabetes implies the existence of a completed causal pathway of predisposing factors. The purpose of this descriptive retrospective study was to evaluate foot care provided to residents in a 179-bed long-term care facility. The charts of all 17 eligible residents with a diagnosis of diabetes mellitus were reviewed for documentation of assessment and care of their feet. All data were collected by the investigator using the Minimum Data Set (MDS) 2.0 Assessment of Diabetic Foot Care Instrument and a demographic and foot care history instrument. Foot problems were documented for 59% of the participants. Podiatrist-documented foot examination was found in only one of the charts reviewed; however, six residents had been referred to podiatrist. Throughout the charts reviewed, no documentation of protective sensation using the Semmes Weinstein monofilament or vibration test was found. The findings of this study are consistent with previous research that showed a gap exists between the established standard and the degree to which the standards are met (Fain & Melkus, 1994; Wylie-Rosett et al., 1995). Adequate attention to the problem by health care providers, efforts to increase awareness of foot care standards, and early intervention may be steps to close the gap. Nurses must identify patients at risk for foot problems and actively intervene to prevent complications from occurring. PMID- 15109043 TI - Pneumonia in the long-term care setting: etiology, management, and prevention. AB - 1.Nursing-home acquired pneumonia (NHAP) is a major cause of death and disability among elderly nursing home residents, despite the availability of new antimicrobials and diagnostic techniques. 2. Elderly individuals with NHAP have vague clinical presentations and unique institutional limitations can lead to delays in diagnosis, treatment, and poor resident outcomes. 3. Successful management of the resident with pneumonia includes choice of antibiotic therapy, excellent nursing care, and thoughtful consideration of treatment setting. 4. Preventive strategies to reduce the risk of NHAP include attention to vaccination status and oral hygiene care to reduce bacterial colonization of potential respiratory pathogens. PMID- 15109044 TI - Medication practices among Chinese American older adults influences: a study of cultural. AB - The magnitude and intensity of medication use in older individuals have been widely discussed. Yet, a paucity of research data exists on medication practices among community-dwelling Chinese American older adults. This study examined factors associated with medication practices in community-dwelling Chinese American older adults. Data were collected through a descriptive survey. Chinese American older adults (n = 165) from the greater Boston area participated in the survey. The study revealed that ethnocultural barriers and a lack of health insurance significantly influenced medication practices among Chinese American older adults. The study suggested three important implications. First, more ethnoculturally appropriate health services are needed. Secondly, communication between clients and health providers must be improved. Finally, culturally sensitive medication management programs are needed. PMID- 15109045 TI - Gambling behaviors in Black older adults: perceived effects on health. AB - Gambling is a popular activity among older adults of multiple ethnic and cultural backgrounds, but gambling research has not focused on elderly individuals or on minority groups. Although most gambling is a social activity, it also could have health implications for older adults. Sixty-seven Black individuals, ages 50 to 88, completed the Massachusetts Gambling Screen and 20 Black older adults were interviewed about gambling behaviors and beliefs about gambling and health. Although participants in this study were low income, they were as active in gambling as the general population. One pathological gambler and two problem gamblers were identified. The correlation between reported overall health and beliefs about the effects of gambling on health was not significant. Researchers, educators, and nurses working with older adults can help define the health implications of gambling on this population. PMID- 15109046 TI - Informed consent: a process to facilitate older adults' participation in research. AB - Trust is the foundation of the informed consent process. According to Kass et al., Yet only through vigilance and humility will we, as investigators, be able to live up to the trust that is placed in us; and only if that trust is deserved can the research enterprise survive (1996, p. 28). Therefore, further research should examine the effect of the informed consent process in limiting older adults' participation. Only by re-evaluating the first point of contact, implementing creative strategies to overcome barriers to the informed consent process, and validating these strategies through research will nurses facilitate the inclusion of older adults in research. PMID- 15109047 TI - Promoting exercise behavior in older adults: using a cognitive behavioral intervention. AB - Health benefits associated with exercise are only obtained when exercise is maintained. The purpose of this pilot study was to examine the effects of a cognitive behavioral therapy (CBT) intervention on exercise behavior and physical outcomes in older adults. All participants were taught progressive flexibility, strength, and endurance exercises. The control group received no additional treatment. The experimental group was taught to recognize negative thoughts related to exercise and to counter these thoughts with more positive ones. Subsequent exercise behavior and physical outcomes were measured in all participants. Cognitive behavioral therapy was moderately to largely effective in improving the majority of physical outcomes assessed. In addition, CBT was moderately effective in improving several components of self-reported exercise behavior and mildly effective in improving exercise behavior overall. Results suggest that nurses can train older adults to identify and modify thoughts that interfere with or reduce their exercise behavior and thus improve physical functioning. PMID- 15109048 TI - Components of feed efficiency in broiler breeding stock: the use of fasted body temperature as an indicator trait for feed conversion in broiler chickens. AB - Five trials were conducted to evaluate the usefulness of body temperature (BT) measured under conditions of mild handling stress as an indicator trait for feed conversion ratio (FCR) in broilers. The trials used males from 4 lines of broilers (a randombred control population and 3 heavy broiler lines). Birds were reared to approximately 6 wk of age and then selected for FCR testing based upon BW and conformation. At the start of the FCR test, birds were fasted for periods lasting from 18 to 48 h, and their BT were measured, which was followed immediately by initiation of an FCR test lasting 1 wk. The correlation between FCR and BT was significant in only 2 of the 5 trials. Differences between good and poor converters were generally attributed to greater gains of BW on similar amounts of feed intake. The results of the current trials indicate gain on test is the primary factor affecting FCR and that BT is not a reliable predictor of FCR. PMID- 15109049 TI - Estimation of genetic parameters for fat deposition and carcass traits in broilers. AB - Abdominal and subcutaneous fat are regarded as the main sources of waste in the slaughterhouse. Fat stored intramuscularly is regarded a favorite trait related to meat quality. The objective of current study was to estimate genetic parameters for fat deposition in the 3 different parts of body and their relationships with other carcass traits. Traits were recorded for 1,752 females and 1,526 males from a meat-type chicken line. Heritability estimates for abdominal fat percentage, skin percentage as a measure of subcutaneous fat, and intramuscular fat percentage were 0.71, 0.24, and 0.08, respectively. Heritabilities of the other carcass traits were moderate to high (0.28 to 0.73). There was a high genetic correlation between abdominal fat weight and skin weight (0.54), whereas the genetic correlation between abdominal fat weight and intramuscular fat percentage was almost zero (0.02). The BW at 7 wk showed a positive genetic correlation with fat production traits, which were high for intramuscular fat percentage (0.87) and moderate for skin percentage (0.17) and abdominal fat percentage (0.13). Therefore carcass traits could be improved by selection for increased breast muscle and reduced abdominal fat without decreased intramuscular fat. PMID- 15109050 TI - Effect of age of feed restriction and microelement supplementation to control ascites on production and carcass characteristics of broilers. AB - Three experiments were conducted, from January until September 2001, to estimate the optimized age to apply feed restriction to control mortality from ascites, with no negative effects on production and carcass characteristics of broilers. For each experiment, 1,200 1-d-old mixed Ross x Peterson chicks were reared in floor pens (50 chicks in each) and fed commercial feed. Feed restriction was applied for 8 h/d for 14 d at 21 or 28 d of age in experiment 1, 14 or 21 d in experiment 2, and 7 or 14 d in experiment 3. In experiments 2 and 3, a microelement supplement (without or with) was tested; the control groups received feed ad libitum and no supplement. Body weight gain, feed conversion, total mortality, and mortality from ascites, leg problems, and carcass characteristics were considered at the end of each experiment. The data were analyzed as a completely randomized design, or as a 2 x 2 factorial to estimate main and interaction effects (experiments 2 and 3). Additional analyses, including the control, were done; means comparisons were by orthogonal contrasts. The production and carcass characteristics of the restricted groups were lower than the control but were not statistically different in experiments 2 and 3, although the optimized age for feed restriction was at 7 d. Total mortality and mortality from ascites decreased by restriction, but leg problems increased without supplement. The results indicated that quantitative feed restriction and microelement supplementation at 7 d of age reduced mortality from ascites and leg problems and permitted compensatory growth sufficient to equal the production characteristics of the control group at 49 d of age. However, it is necessary to determine the specific microelements to be supplemented and to estimate the effects of season and genetic line. PMID- 15109051 TI - Effects of feeding blends of grains naturally contaminated with Fusarium mycotoxins on growth and immunological parameters of broiler chickens. AB - An experiment was conducted to investigate the effects of feeding grains naturally contaminated with Fusarium mycotoxins on growth and immunological parameters of broiler chickens. Three hundred sixty, 1-d-old male broiler chicks were fed 1 of 4 diets containing grains naturally contaminated with Fusarium mycotoxins for 56 d. The diets included (1) control; (2) low level of contaminated grains (5.9 mg/kg deoxynivalenol (DON), 19.1 mg/kg fusaric acid (FA), 0.4 mg/kg zearalenone, and 0.3 mg/kg 15-acetyldeoxynivalenol; (3) high level of contaminated grains (9.5 mg/kg DON, 21.4 mg/kg FA, 0.7 mg/kg zearalenone, and 0.5 mg/kg 15-acetyldeoxynivalenol); and (4) high level of contaminated grains + 0.2% polymeric glucomannan mycotoxin adsorbent (GM polymer). Body weight gains and feed consumption of chickens fed contaminated grains decreased linearly with the inclusion of contaminated grains during the grower phase (d 21 to 42). Efficiency of feed utilization, however, was not affected by diet. Production parameters were not significantly affected by the supplementation of GM polymer to the contaminated grains. Peripheral blood monocytes decreased linearly in birds fed contaminated grains. The feeding of contaminated diets linearly reduced the B-cell count at the end of the experiment, whereas the T-cell count on d 28 responded quadratically to the contaminated diets. The feeding of contaminated diets did not significantly alter serum or bile immunoglobulin concentrations, contact hypersensitivity to dinitrochlorobenzene, or antibody response to SRBC. Supplementation with GM polymer in the contaminated diet nonspecifically increased white blood cell count and lymphocyte count, while preventing mycotoxin-induced decreases in B-cell counts. It was concluded that broiler chickens are susceptible during extended feeding of grains naturally contaminated with Fusarium mycotoxins. PMID- 15109052 TI - Analysis of chicken TLR4, CD28, MIF, MD-2, and LITAF genes in a Salmonella enteritidis resource population. AB - Salmonella enteritidis is a foodborne pathogen that negatively affects both animal and human health. Genetic variations in response to pathogenic SE colonization or to SE vaccination were measured in a chicken resource population. Outbred broiler sires and 3 diverse, highly inbred dam lines produced 508 F1 progeny that were evaluated for either bacterial colonization after pathogenic SE inoculation or circulating antibody level after SE vaccination. Five candidate genes were selected for study, based on their biological function as possibly affecting response to SE: toll-like receptor 4 (TLR4), T-cell specific surface protein (CD28), macrophage migration inhibitory factor (MIF), MD-2, and lipopolysaccharide-induced tumor necrosis factor (TNF)-alpha factor (LITAF). Gene fragments were sequenced from the founder lines of the resource population. The LITAF and MIF genes were homozygous for all sires. Single nucleotide polymorphisms (SNP) were identified in 3 genes (TLR4, CD28, and MD-2) and were used to test for associations of sire SNP with SE response. Linear mixed models were used for statistical analyses. The CD28 broiler sire SNP was associated with both bacterial load in the cecum (P < 0.003) and vaccine antibody response (P < 0.05). The MD-2 SNP was associated (P < 0.04) with the bacterial load in the spleen. The use of these SNP in these genes in marker-assisted selection may result in enhancement of disease resistance. PMID- 15109053 TI - Proteomics in the chicken: tools for understanding immune responses to avian diseases. AB - The entire chicken genome sequence will be available by the time this review is in press. Chickens will be the first production animal species to enter the "postgenomic era." This fundamental structural genomics achievement allows, for the first time, complete functional genomics approaches for understanding the molecular basis of chicken normo- and pathophysiology. The functional genomics paradigm, which contrasts with classical functional genetic investigations of one gene (or few) in isolation, is the systematic holistic genetic analyses of biological systems in defined contexts. Context-dependent gene interactions are the fundamental mechanics of all life. Functional genomics uses high-throughput large-scale experimental methods combined with statistical and computational analyses. Projects with expressed sequence tags in chickens have already allowed the creation of cDNA microarrays for large-scale context-dependant mRNA analysis (transcriptomics). However, proteins are the functional units of almost all biological processes, and protein expression very often bears no correlation to mRNA expression. Proteomics, a discipline within functional genomics, is the context-defined analysis of complete complements of proteins. Proteomics bridges the "sequence-to-phenotype gap;" it complements structural and other functional genomics approaches. Proteomics requires high capital investment but has ubiquitous biological applications. Although currently the fastest-growing human biomedical discipline, new paradigms may need to be established for production animal proteomics research. The prospective promise and potential pitfalls of using proteomics approaches to improve poultry pathogen control will be specifically highlighted. The first stage of our recently established proteomics program is global protein profiling to identify differentially expressed proteins in the context of the commercially important pathogens. Our trials and tribulations in establishing our proteomics program, as well some of our initial data to understand chicken immune system function, will be discussed. PMID- 15109054 TI - Our current understanding of humoral immunity of poultry. AB - With the inception of the modem era of avian immunology in the mid-1950s following the discovery of the role of the bursa of Fabricius (BF), steady and persistent research has been conducted by scientists. Findings that have advanced our knowledge of humoral immunity (HI) include the distinct role of B cells in antibody production, explanation of antibody diversity by gene rearrangement and conversion, identification of a bursal secretory dendritic cell and dendritic cells of the spleen, characterization of cytokine factors, and mapping of MHC class II genes. Numerous experiments have been conducted to evaluate the HI responses of poultry through antibody titer determination and quantitation of immunoglobulin concentrations in the humors. Antibody studies have included the genetics of HI regarding both quantity and persistence of responses as influenced by multiple genes and MHC haplotypes. In the past decade, emergence of monoclonal antibody reagents to identify cell surface markers on B cells and accessory cells has allowed for discrimination of HI functions of cells by immunohistochemistry, flow cytometry, and other techniques. The signaling mechanisms of avian B cells are becoming better understood. Continued work in avian immunity has led to more advances in identification, characterization, and gene sequencing of important cytokines. Additional work is required to identify and sequence genes for many more cytokines that have direct effects on B cells. A brief history of research published on avian B cells is presented with an examination of some of the important research findings reported in recent years. PMID- 15109055 TI - Cell-mediated immunity in poultry. AB - In avian species, adaptive immunity involves both humoral and cell-mediated immune (CMI) responses. Although humoral or antibody-mediated immune responses are particularly effective against extracellular antigens, CMI responses are specialized in the elimination of intracellular antigens; the latter include those that have entered cells via the endocytic pathway (exogenous antigens; e.g., phagocytosed bacteria) or were produced within the cell such as viral proteins and proteins resulting from neoplastic transformation of the cell (endogenous antigens). CMI responses, like most humoral immune responses, are tightly regulated and require "help" from T helper cells, specifically the type 1 T helper cells (Th1, hence, the name Th1 responses). Th1 cells are characterized by their production of cytokines such as interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and interleukin-2 that drive CMI responses. The functional effectors of CMI responses are various immune cells including cytotoxic lymphocytes (cytotoxic T cells and natural killer cells) and macrophages. Cytotoxic lymphocytes and macrophages are specialized in the elimination of endogenous and exogenous antigens, respectively. In the past decade, substantial progress has been made in defining the role and regulation of avian CMI responses. Other advances have addressed strategies that strengthen this arm of adaptive immunity to optimize defense as well as protection against neoplastic diseases and nonneoplastic diseases caused by intracellular pathogens. PMID- 15109056 TI - Participation of the intestinal epithelium and mast cells in local mucosal immune responses in commercial poultry. AB - The intestinal mucosa of commercial poultry is continually subjected to invasion or colonization by a wide array of potentially hostile enteric pathogens. Although, recent investigations have focused on lymphocyte involvement in immune responses in the intestine, lymphocyte-mediated immunity alone will not explain the barrier nature of mucosal membranes associated with rejection of many enteric pathogens upon secondary homologous challenge. Our laboratories have focused on nontraditional elements of mucosal immunity in poultry to better understand host pathogen interactions in the intestine. Following classical and novel immunization procedures, we have identified an antigen-specific mechanism of immediate responsiveness of the mucosal epithelium characterized by epithelial chloride secretion. This mechanism, characteristic of intestinal anaphylaxis, is mediated by local immune elements. Similar mechanisms in mammals contribute to the barrier nature of mucosal membranes during pathogen challenge. To identify cells participating in these and similar responses, additional studies have described a role for mast cells in acute phase responses in the intestines of chickens experimentally challenged with Eimeria. To a more practical end, other experiments in our laboratories have characterized drinking water administration of BSA for elicitation of local and systemic antibody responses. These experiments have shown ad libitum drinking water administration of BSA to be as effective as i.p. administration of BSA; they present a novel approach to immunization of commercial poultry with protein vaccines. These investigations support continued research on host-pathogen interactions within the intestine of commercial poultry to better understand and control enteric pathogens through vaccination or immunomodulation. PMID- 15109057 TI - Distinctive polymorphism of chicken B-FI (major histocompatibility complex class I) molecules. AB - The major histocompatibility complex (MHC) in chickens influences disease resistance, but the mechanism is not understood. In Leghorn lines, the MHC contains 2 closely-linked class I loci, B-FI and B-FIV. Previously, we determined nucleotide sequences of well-expressed class I (B-F) genes from unique MHC haplotypes of broiler chicken lines. More recently, we identified 7 new B-F alpha1alpha2-coding sequences from less well-expressed loci by amplification of genomic DNA from unique broiler haplotypes. Phylogenetic analysis of chicken MHC class I alpha1alpha2-coding sequences resolved 2 clusters (Groups A and B), which appear to correspond to B-FIV and B-FI loci, respectively. Compared with B-FIV locus, B-FI alleles were less polymorphic overall, but nevertheless demonstrated evidence of diversifying selection. The most striking feature of B-FI alleles is a conserved, locus-specific motif in the alpha helix of the alpha1 domain, a region that is highly variable in B-FIV alleles. This distinctive pattern of allelic polymorphism resembles that of the HLA-C class I locus in the human MHC (HLA). The conservation of the alpha helix of the alpha1 domain relates to HLA-C interaction with members of the killer immunoglobulin-like receptors on natural killer (NK) cells that are specific for recognition of HLA-C molecules and function to regulate activation of NK cells. Whereas HLA-C molecules may be dominant ligands for NK cell regulation, HLA-A and -B molecules are more important in presenting antigen to cytotoxic T lymphocytes. We hypothesize that chicken B-FI molecules may be specialized to serve similar functions as HLA-C molecules. PMID- 15109058 TI - Non-major histocompatibility complex alloantigen genes affecting immunity. AB - An alloantigen is a genetically determined cell-surface molecule detected by specific antisera. An identifying letter has been assigned to each genetic locus responsible for the 12 distinct families of alloantigens: A, B, C, D, E, H, I, J, K, L, P, and R. The genes of each system segregate independently of the other systems, except that the A and E are very closely linked (0.5 centimorgans). Selection experiments over numerous generations have revealed distinct changes in gene frequency of the A-E alloantigens, suggesting immune responses associated with susceptibility to coccidiosis, response to immunizations with SRBC, and selection for size of the bursa of Fabricius. Immune response effects of the C system of alloantigen genes are indicated by distinct gene frequency changes following selection for response to SRBC, selection for size of bursa of Fabricius, and macrophage nitrite production after lipopolysaccharide (LPS) stimulation. Immune response effects of the D system of antigens are indicated by data from genetic selection for response to immunization with SRBC, selection for bursa size, and macrophage nitrite and cytokine interleukin (IL)-6 production following LPS stimulation. Immune response effects of the I system genes are indicated by distinct gene frequency changes in lines selected for bursa size and within family comparisons for macrophage nitrite and cytokine IL-6 production following LPS stimulation. Effects of the L system, consisting of only 2 alleles, are indicated by the gene frequency changes following selection for bursa size, direct comparison of genotypes within families for monocyte phagocytosis, susceptibility to coccidiosis, outcome of Rous sarcomas, and immune responses to SRBC and Brucella abortus. Genotypes of the P alloantigen system were directly compared within families of fully pedigreed chicks with significant differences for monocyte phagocytosis. An experimental procedure for simultaneously testing for immune responses of genotypes of 9 of the alloantigen systems (A, B, C, D, E, H, I, L, and P) has been established by producing test progeny from a single cross of parent lines segregating for genes of each of the systems. PMID- 15109059 TI - Recent progress on the cytokine regulation of intestinal immune responses to Eimeria. AB - A variety of methods are available to combat avian diseases in the commercial setting, including improved farm management practices, use of antibiotic drugs, selection of disease resistant chicken strains, and manipulation of the chicken immune system. In the latter category, development of vaccines against the major avian diseases has become a priority for the poultry industry. With increasing demands for developing alternative control programs for many poultry diseases, it is important to understand the basic immunobiology of host-pathogen interactions in order to develop novel vaccination strategies. From studies carried out in many mammalian species, it is evident that host immune responses to intracellular pathogens are complex and involve many components of the host immune system. For enteric pathogens such as Eimeria and Salmonella, understanding cell-mediated immunity is most important because antibodies, although abundantly produced locally, can not access and act on these intracellular pathogens. In poultry, slow but increasing understanding of various components of host immune system mediating cellular immunity is opening new opportunities for thorough investigation of the role of thymus-derived lymphocyte subpopulations and cytokines in normal and disease states. This paper will review recent progress with chicken cytokines that have been characterized, and discuss various experimental strategies to enhance host immunity to pathogens using chicken cytokines. PMID- 15109060 TI - Immune modulation of the pulmonary hypertensive response to bacterial lipopolysaccharide (endotoxin) in broilers. AB - The lungs of broilers are constantly challenged with lipopolysaccharide (LPS, endotoxin) that can activate leukocytes and trigger thromboxane A2 (TxA2)- and serotonin (5HT)-mediated pulmonary vasoconstriction leading to pulmonary hypertension. Among broilers from a single genetic line, some individuals respond to LPS with large increases in pulmonary arterial pressure, whereas others fail to exhibit any response to the same supramaximal dose of LPS. This extreme variability in the pulmonary hypertensive response to LPS appears to reflect variability in the types or proportions of chemical mediators released by leukocytes. Our research has confirmed that TxA2 and 5HT are potent pulmonary vasoconstrictors in broilers and that broilers hatched and reared together consistently exhibit pulmonary hypertension after i.v. injections of TxA2 or 5HT. Previous in vitro studies conducted using macrophages from different lines of chickens demonstrated innate variability in the LPS-stimulated induction of nitric oxide synthase (iNOS) followed by the onset of an LPS-refractory state. The NOS enzyme converts arginine to citrulline and nitric oxide (NO). It is known that NO produced by endothelial NOS serves as a key modulator of flow-dependent pulmonary vasodilation, and it is likely that NO generated by iNOS also contributes to the pulmonary vasodilator response. Accordingly, it is our hypothesis that the pulmonary hypertensive response to LPS in broilers is minimal when more vasodilators (NO, prostacyclin) than vasoconstrictors (TxA2, 5HT) are generated during an LPS challenge. Indeed, inhibiting NO production through pharmacological blockade of NOS with the inhibitor Nomega-nitro-L-arginine methyl ester modestly increased the baseline pulmonary arterial pressure and dramatically increased the pulmonary hypertensive response to LPS in all broilers evaluated. Innate differences in the effect of LPS on the pulmonary vasculature may contribute to differences in susceptibility of broilers to pulmonary hypertension syndrome (ascites). PMID- 15109061 TI - Major histocompatibility (B) complex control of responses against Rous sarcomas. AB - The chicken major histocompatibility (B) complex (MHC) affects disease outcome significantly. One of the best characterized systems of MHC control is the response to the oncogenic retrovirus, Rous sarcoma virus (RSV). Genetic selection altered the tumor growth pattern, either regressively or progressively, with the data suggesting control by one or a few loci. Particular MHC genotypes determine RSV tumor regression or progression indicating the crucial B complex role in Rous sarcoma outcome. Analysis of inbred lines, their crosses, congenic lines, and noninbred populations has revealed the anti-RSV response of many B complex haplotypes. Tumor growth disparity among lines identical at the MHC but differing in their background genes suggested a non-MHC gene contribution to tumor fate. Genetic complementation in tumor growth has also been demonstrated for MHC and non-MHC genes. RSV tumor expansion reflects both tumor cell proliferation and viral replication generating new tumor cells. In addition, the B complex controls tumor growth induced by a subviral DNA construct encoding only the RSV v-src oncogene. Immunity to subsequent tumors and metastasis also exhibit MHC control. Genotypes that regressed either RSV or v-src DNA primary tumors had enhanced protection against subsequent homologous challenge. Regressor B genotypes had lower tumor metastasis compared with progressor types. Together, the data indicate that B complex control of RSV tumor fate is strongly defined by the response to a v-src-determined function. Differential RSV tumor outcomes among various B genotypes may include immune recognition of a tumor-specific antigen or immune system influences on viral replication. PMID- 15109062 TI - Nutritional modulation of immune function in broilers. AB - Collaborative research efforts across disciplines typically result in more insight toward the hypothesis being tested due to the omnibus nature of the projects. For example, nutritional experiments evaluating a nutrient response will benefit greatly by incorporating biochemical, physiological, and immunological endpoints for measurement. Clearly, commercial poultry producers do not have the luxury of focusing on specific disciplines when field problems occur. Hence, in practice interplay exists among nutrition, genetics, management, and diseases. Dietary composition impacts immune function of the chicken. As research in the area of nutritional immunology has increased, it is becoming apparent that nutrient needs for immunity do not coincide with those for growth or skeletal tissue accretion. This review is not a comprehensive assessment of nutrient needs for immunity in the chicken. Rather, this review is concerned with nutritional modulation of immunity in broilers that offers insight for nutritionists and researchers to implement nutritional regimens to reduce the severity of disease and to test or validate nutritional regimens that heighten immunity. Nutritional modulation of the hen diet and in ovo nutrient modulation to improve chick immunity and disease resistance are discussed. PMID- 15109063 TI - Selection for avian immune response: a commercial breeding company challenge. AB - Selection for immune function in the commercial breeding environment is a challenging proposition for commercial breeding companies. Immune response is only one of many traits that are under intensive selection, thus selection pressure needs to be carefully balanced across multiple traits. The selection environment (single bird cages, biosecure facilities, controlled environment) is a very different environment than the commercial production facilities (multiple bird cages, potential disease exposure, variable environment) in which birds are to produce. The testing of individual birds is difficult, time consuming, and expensive. It is essential that the results of any tests be relevant to actual disease or environmental challenge in the commercial environment. The use of genetic markers as indicators of immune function is being explored by breeding companies. Use of genetic markers would eliminate many of the limitations in enhancing immune function currently encountered by commercial breeding companies. Information on genetic markers would allow selection to proceed without subjecting breeding stock to disease conditions and could be done before production traits are measured. These markers could be candidate genes with known interaction or involvement with disease pathology or DNA markers that are closely linked to genetic regions that influence the immune response. The current major limitation to this approach is the paucity of mapped chicken immune response genes and the limited number of DNA markers mapped on the chicken genome. These limitations should be eliminated once the chicken genome is sequenced. PMID- 15109064 TI - An evaluation of endo-beta-D-mannanase (Hemicell) effects on broiler performance and energy use in diets varying in beta-mannan content. AB - Two experiments were conducted to evaluate the effects of a commercial endo-beta D-mannanase (Hemicell) on overall performance, MEn, net energy for gain, and some serum parameters of broilers fed diets varying in beta-mannan level (experiment 1) and to evaluate effects of enzyme level on the same variables in broilers fed diet high in beta-mannan (experiment 2). As a semipurified beta-mannan source, guar gum was used to alter the dietary beta-mannan level. In experiment 1, guar gum was added at 0, 0.5, 1, and 2% in a corn-soy-based starter diet with (0.05%) and without endo-beta-D-mannanase supplementation in a 4 x 2 factorial design. Enzyme supplementation improved (P < 0.01) feed efficiency at control and each guar gum inclusion level, whereas 2% guar gum supplementation reduced (P < 0.01) BW and increased (P < 0.01) 14-d feed:gain ratio. Enzyme supplementation also increased dietary MEn and net energy gain. In experiment 2, endo-beta-D-mannanase was added at 0, 0.5, 1, and 1.5% in a corn-soy-based starter diet containing 1% guar gum. Increasing endo-beta-D-mannanase supplementation did not affect (P > 0.10) final BW but improved 14-d feed:gain ratio at all inclusion levels. As in the first experiment, ME improved (P < 0.05) with increasing enzyme inclusion. Dietary endo-beta-D-mannanase inclusion significantly reduced water:feed ratio and total dry fecal output (P < 0.01). Taken together, the results of these 2 experiments indicate that endo-beta-D-mannanase supplementation may improve the utilization of nutrients in diets containing beta-mannan. PMID- 15109065 TI - The effect of two different blends of essential oil components on the proliferation of Clostridium perfringens in the intestines of broiler chickens. AB - The effect of 2 different blends of essential oils on Clostridium perfringens (Cp) in the intestine and feces of broiler chickens was tested in 6 field trials for each blend. One hundred parts per million of the blends were mixed in a commercial corn-based diet throughout the entire growing period for experimental flocks. Samples from the jejunum, cecum, cloaca, and feces were taken on d 14, 21, and 30 from experimental and control flocks and tested quantitatively for Cp via blood agar plate, litmus milk medium, and ELISA. Blend A reduced (P < or = 0.05) the average Cp concentration in the feces on all sampling days, in the jejunum and cecum on d 14 and 21, and in the cloaca on d 14. Blend B effected a significant reduction of Cp concentration in the jejunum on d 14 and 30 and in the cloaca on d 14. The percentages of specimens from the control group that tested positive for Cp were 83.3% for feces, 88.0% for jejunum and cloaca, and 82.6% for cecum. Specimens from the feces and 3 sections of the intestine were Cp positive in groups treated with blend A (60.8, 64.6, 47.9, and 70.8%) and with blend B (65.9, 63.6, 63.6, and 72.7%). Our results indicate that specific blends of essential oil components can control Cp colonization and proliferation in the gut of broilers and therefore may be of help to prevent problems with Cp and necrotic enteritis. PMID- 15109066 TI - Growth performance and nitrogen excretion of broilers using a phase-feeding approach from twenty-one to sixty-three days of age. AB - Two experiments were conducted to assess effects of phase-feeding (PF) on broilers from 21 to 63 d. Experiment 1 evaluated the impact of PF on growth performance, whereas experiment 2 assessed the effects of PF on CP intake and nitrogen excretion. Diets were formulated using recommendations from NRC or linear regression equations. Two PF treatments were prepared: standard (PF) and low (PF10), in which predicted Lys, sulfur amino acid, and Thr recommendations were reduced by 10%. For PF and PF10, 2 diets (high-nutrient and low-nutrient density) were blended in variable quantities to produce rations matching predicted requirements. An NRC grower and finisher diet or a series of PF and PF10 diets that were switched every other day were fed. In experiment 1, weight gain and feed efficiency were improved (P < 0.05) by PF10 relative to broilers fed the NRC-based diet. Crude protein intake was reduced (P < 0.05) by PF10 relative to broilers fed NRC and PF diets. No differences (P > 0.05) in percentage carcass composition were observed when broilers were fed PF or PF10 diets. Significant reductions (P < 0.05) in dollars per kilogram of weight gain were noted with PF regimens. In experiment 2, PF and PF10 diets reduced (P < 0.05) CP intake and nitrogen excretion from 43 to 63 d. Results indicate that PF regimens may substantially reduce dietary costs and may have environmental benefits. PMID- 15109067 TI - True amino acid availability in chinese high-oil corn varieties determined in two types of chickens. AB - True amino acid availability (TAAA) was determined in conventional corn (CC) and in 2 Chinese high-oil corn varieties (CHOC1; CHOC2). The CC, CHOC1, and CHOC2 contained 3.7, 6.6, and 7.5% oil (ether extract) on a dry matter basis, respectively. The CP content (% dry matter) of the corn increased from 8.6 to 12.2% as oil increased from 3.7 to 7.5%. Birds (Hy-Line roosters or Arbor Acres broilers) were given 1 of the 3 corn varieties as the sole dietary ingredient in a conventional (intubation method) TAAA assay. There was a significant (P < 0.05) effect of bird type on gut metabolic (endogenous) amino acid excretion values with the broilers having higher excretions. For all amino acids, there was also a significant (P < 0.05) effect of bird type on TAAA with the Hy-Line roosters having higher TAAA values. The mean value for TAAA calculated over all of the amino acids showed a 9.8% unit higher digestibility for roosters compared with the broilers. Differences in TAAA between the corn varieties were statistically significant for alanine and valine only. Overall, the present results indicate that TAAA is equal to or superior to that in CC and that the TAAA content is higher in CHOC compared with CC. Bird type has an important effect on TAAA results and should be considered when developing TAAA assays and interpreting TAAA values. PMID- 15109068 TI - Benefit of feeding dietary calcium and nonphytate phosphorus levels above National Research Council recommendations to tom turkeys in the growing-finishing phases. AB - This experiment evaluated the effects of feeding various dietary Ca and nonphytate P (nPP) levels to Large White male turkeys from 3 to 17 wk of age. After consuming a common prestarter diet, poults were fed approximate NRC (1994) levels of dietary Ca and nPP from 3 to 9 wk of age or levels approximately 25% higher. From 9 to 17 wk of age, each starter group was fed approximately 75 (low P), 100 (medium P), or 145% (high P) of the NRC (1994) requirements for Ca and nPP. Diets were fed as crumbles to 6 wk of age and as pellets from 6 to 17 wk of age. There were no effects on BW or feed efficiency to 9 wk of age. Litter P was increased by 21% when high Ca and nPP were fed from 3 to 9 wk. High dietary Ca and nPP fed during the growing-finishing period generally improved bone strength and ash. Tibia strength and ash were higher in the medium P group compared with in the low P group. Wing bone strength was greater in the high P group than in both the birds fed low or medium P. Litter P was increased by 23% when High P was fed in the growing-finishing period compared to the birds fed the Medium P diet during the same period. The results show there is a benefit to bone strength and mineralization when Ca and nPP are fed at levels higher than NRC (1994) recommendations. PMID- 15109069 TI - Noninvasive characterization of the turkey heart performance and its relationship to skeletal muscle volume. AB - The cardiovascular capacity of turkeys is considerably affected by the selection procedure for meat production. To determine the body weight-related changes of some quantitative characteristics of the turkey heart function, serial measurements were carried out. BUT Big 6 turkeys (both sexes) were examined using whole-body magnetic resonance imaging (MRI) and spiral computer tomography (CT). Based on dynamic MRI examinations the following significant age- and sex dependent quantitative differences were found. Means of the left ventricular stroke volumes were measured at the ages of 12 wk (males, 4.18 mL; females 2.89 mL), 16 wk (males, 6.56 mL; females, 4.14 mL), and 20 wk (males, 8.17 mL; females, 5.16 mL). Average cardiac output (CO) values were 0.76, 1.14, and 1.33 L/min for males and 0.58, 0.79,0.93 L/min for females. A statistically unproven age-dependent increasing tendency in the estimated relative CO value (skeletal muscle tissue volume measured by CT/CO unit) of male and female turkeys was found under sedentary conditions. These changes in the relative CO value and also the determined decrease of CO value related to body surface may, in theory, indicate a disadvantageous process that is more pronounced in males. The combined application of CT and MRI can be a powerful approach for studying the relationship between skeletal muscle development and heart capacity. PMID- 15109070 TI - The role of sodium ions in the pathogenesis of skeletal muscle damage in broiler chickens. AB - The effect of sodium ions (Na+) on calcium (Ca2+)-mediated muscle damage in broiler chickens was investigated using an in vitro muscle preparation. Muscle Ca2+ accumulation was determined by 45Ca2+ uptake. Muscle damage was assessed by measurement of the efflux of the intracellular enzyme creatine kinase (CK) into the incubation medium. Loading muscle cells with Na+ by means of the sodium ionophore monensin led to concentration-dependent (25 to 200 microM) increases in 45Ca2+ uptakes and corresponding and proportional CK losses. The greatest responses occurred at 100 microM ionophore or greater, reflected in a 49% increase (P < 0.05) in 45Ca2+ uptake and an associated 140%-fold increase (P < 0.001) in CK efflux. Inhibition of muscle Na+/K+-ATPase activity with ouabain (2 mM) induced a 56% increase in 45Ca2+ uptake and a 60%-fold increase (P < 0.001) in total CK loss. The combined use of ionophore and ouabain resulted in 90 and 130%-fold elevations in 45Ca2+ uptake and CK loss, respectively. In monensin treated muscles, inhibition of external Ca2+ influx from the incubation medium by chelation with 1,2 bis(2-aminophenoxy)ethane-N,N,N',N' tetracetic acid (5 mM) markedly reduced 45Ca2+ uptake (38%: P < 0.05) but increased CK release by 85% (P < 0.001). The results demonstrate that initial elevations in muscle Na+ can facilitate increases in muscle Ca2+ and lead to alterations in muscle cell membrane integrity and CK loss. The Na+-induced increases in myocellular Ca2+ may be mediated via direct extracellular Ca2+ entry or redistribution from internal Ca2+ stores. It is proposed that in order to reduce or prevent myopathies in poultry, exposure to conditions that may lead to elevations in muscle Na+ (e.g., increased muscle activity and stress or accidental ionophore toxicosis) should be avoided. The findings of this study have implications for management strategies of bird welfare, muscle pathology, and product quality. PMID- 15109071 TI - Hyaluronan and lymphedema. PMID- 15109072 TI - Trafficking of hyaluronan in the interstitium and its possible implications. AB - The mechanisms underlying the pathological changes in lymphedematous tissue are far from clear, and it is becoming apparent that plasma proteins may not be the only key factors responsible for holding water in the interstitium. This review focuses on an "old" macromolecule--hyaluronan (HA) which is one of the major components of the interstitium and has a close relationship with the lymphatic system. Growing recognition of the multiple functions of this macromolecule for important physiological and pathological events may be helpful in identifying the crucial changes in tissues subjected to lymphatic circulation insufficiency and ultimately in the search for rational therapeutic approaches to prevent or reverse these tissues changes. PMID- 15109073 TI - Micropatterned hyaluronan surfaces promote lymphatic endothelial cell alignment and orient their growth. AB - The implant of a biocompatible device capable of guiding lymphatic vessel regeneration in patients who underwent removal of lymph nodes might contribute to restoring an efficient lymphatic drainage and help to prevent the occurrence of lymphedema. The aim of this study was to evaluate whether a microstructured surface could provide a guidance for the growth of cultured lymphatic endothelial cells. The presence of microstructures on a surface permits the control of cell adhesion, migration, proliferation, and differentiation. We report here that lymphatic endothelial cells align on microstructures of alternating hyaluronan and aminosylanized glass stripes obtained by photoimmobilization. Cells consistently spread and proliferate only on aminosylanized glass. They orient parallel to the longitudinal axis of the stripe. A pattern of alternating stripes of aminosylanized glass uniformly covered by elongated cells and of hyaluronan devoid of cells eventuallyforms. The presence of alpha(v)-integrins along cell borders of cells in search of contact with each other and at the leading edge of migrating cells, sites where new focal adhesions are presumably formed, indicates that integrin-mediated adhesion to the substrate guides cell migration along the microstructure. Micropatterned surfaces of hyaluronan thus proved to adequately orient the growth of cells allowing the regeneration of lymphatic endothelium in the desired direction. PMID- 15109074 TI - Multimodal imaging in the congenital pulmonary lymphangiectasia-congenital chylothorax-hydrops fetalis continuum. AB - We report on three infants with congenital chylothorax (CC) and congenital pulmonary lymphangiectasia (CPL). CPL appears to be a characteristic pathological finding in CC. Through the use of lymphoscintigraphy and computed tomography, this study suggests that CC and CPL are strongly correlated entities and that the dysplasia of the lymphatic system results in a pulmonary lymphatic obstruction sequence. The initial microscopic dilatation of the lymph channels may lead to progressive weeping of lymphatics and, consequently, to pleural effusion. Non Immune Hydrops Fetalis (NIHF) may be the final consequence of impaired systemic venous return and may help to explain pleural-pulmonary involvement in this generalized lymph-vessel malformation syndrome. PMID- 15109075 TI - Effect of acetylcholine on in vitro IL-2 production and NK cell cytotoxicity of rats. AB - In the present study, we explored the effect of acetylcholine (ACh) on lymphocyte function and the receptor mechanisms mediating the effect. Concanavalin A (Con A) induced interleukin-2 (IL-2) production and natural killer (NK) cell cytotoxicity were used to assess function of the T lymphocytes and the NK cells from rat spleens. Muscarinic ACh receptors (mA ChRs) agonist pilocarpine and antagonist atropine, as well as nicotinic ACh receptors (nAChRs) agonist nicotine and antagonist tubocurarine were used to determine the action pathways of ACh on T and NK cells. ACh at the concentrations of 10(-10) to 10(-8) M exerted an enhancing effect on Con A-induced IL-2 production and an inhibitory effect on NK cell cytotoxicity. Both pilocarpine and nicotine at the same doses as ACh could mimic these effects of ACh. The enhancing effect of ACh on IL-2 production could be blocked by either atropine or tubocurarine. But the inhibitory effect of ACh on NK cell cytotoxicity was abolished only by atropine, not by tubocurarine. These results suggest that ACh, which is a neurotransmitter of peripheral parasympathetic nervous system, can regulate function of T and NK cells, and the different regulatory effects of ACh on the two types of lymphocytes may be mediated by the different receptor mechanisms. PMID- 15109076 TI - Neurophysiological effects of corticotropin-releasing factor in living slices of the olfactory area of the rat cortex. AB - Application of corticotropin-releasing factor (CRF) at concentrations of 10(-9) and 10(-8) M to living brain slices induced activation of the pre- and postsynaptic excitatory components of focal potentials recorded in the slices. The amplitudes and durations of the AMPA and NMDA components of EPSP increased during exposure to CRF, while the amplitude of the GABA(B)-mediated IPSP was suppressed. At the higher CRF concentration (10(-8) M), cells in slices showed epileptiform discharges. The effects of CRF were reversible and disappeared on washing. Long-term treatment with CRF (90 min) induced changes in cells in slices similar to those associated with long-term post-tetanic potentiation. These data provide evidence that CRF has marked activatory properties and influences the glutamatergic and GABAergic systems. PMID- 15109078 TI - Reflection of an orienting reflex in the phases of evoked potentials in the rabbit visual cortex and hippocampus during substitution of stimulus intensity. AB - Experiments on conscious rabbits were performed using the oddball paradigm, in which a rare (deviant) and common (standard) stimuli were of the same color but different intensities. Deviant stimuli were of lesser intensity. Recordings were made of evoked potentials induced by series of uniform deviant stimuli (without using standard stimuli), which were presented at the beginning and end of stimulation. Visual evoked potentials recorded in response to deviant stimuli in the visual cortex and hippocampus showed increases in the amplitudes of phases, shifted towards positivity as compared with responses to standard stimuli and uniform deviant stimuli at the beginning and end of stimulus blocks. Significant changes affected phases P1 and P2 of visual evoked potentials in the cortex and phases P1, N1, and P2 in the hippocampus. The most significant increase in evoked potentials in the cortex was seen for the P2 peak (P130). It is suggested that changes in responses to oddball-deviant stimuli result from an orienting reflex to rare, unexpected stimuli and that the P2 (P130) peak in the cortex is associated with transmission of information regarding changes in the intensity of the light. The amplitude of this peak was shown to be decreased in responses to uniform deviant stimuli at the beginning and end of stimulus blocks. It was also demonstrated that the clearest and most contrasting changes in visual evoked potentials in responses to deviant and standard stimuli were seen with the smallest differences in intensity between these types of stimulus, this reflecting increases in the orienting reflex at threshold differences. PMID- 15109077 TI - Brain mechanisms for the formation of new movements during learning: the evolution of classical concepts. AB - Current concepts hold that the role of the motor cortex is limited to the control of the appropriate motoneurons on the "point-to-point" principle during the performance of specialized movements of the distal parts of the limbs. However, the last decade has seen the appearance of many data on the plasticity of the motor cortex and its active participation in the process of motor learning. Expression of fos genes has been observed in the motor cortex during the formation of specialized movements. Increases in intracortical horizontal connections in layers II-III during learning fine movements has been seen. The cholinergic input to layers II-III of the motor cortex plays a significant role in this. At the same time, data obtained by functional brain mapping have provided evidence that the activity of the motor cortex also increases during the practice of previously learned movements. This raises the question of the specific function of the motor cortex in the process of motor learning. During the formation of new movements during motor training, a number of previously used synergies interfere with the performance of newly formed coordinations and must be inhibited. The central mechanisms of interference of coordinations in humans have only just started to receive study. At the same time, there is an experimental model for the reorganization and inhibition of interfering synergies in animals. Reorganization of coordinations and inhibition of synergies interfering with the performance of a new movement have been shown to be a specific function of the motor area of the cortex. Cortical control persists during the automation of these synergies, which is not the case in other types of learned movements, though this in itself does not mean that conscious control of their performance also persists. PMID- 15109079 TI - The role of the medial preoptic area of the hypothalamus in organizing the paradoxical phase of sleep. AB - Chronic experiments were performed on cats to study the effects of electrical stimulation of the medial preoptic area of the hypothalamus on the latent period, duration, and structure of paradoxical sleep, as well as the dynamics of neuron activity in this structure during the sleep-waking cycle. These investigations showed that low-frequency stimulation of the medial preoptic area during slow wave sleep led to short-latency development of desynchronization of bioelectrical activity in the neocortex and initiated the development of paradoxical sleep or a similar state. Stimulation of this structure during paradoxical sleep led to a decrease in its duration, to the virtually complete disappearance of the tonic stage of paradoxical sleep, and to an increase in the frequency of rapid eye movements in the phasic stage. Rearrangement of neuron activity in the medial preoptic area during the sleep-waking cycle was similar to that seen in cells of the lower brainstem "executive" centers of paradoxical sleep. It is suggested that neurons in the medial preoptic area of the hypothalamus are actively involved in the mechanisms of paradoxical sleep and, in particular, in the desynchronization of neocortical bioelectrical activity which develops during this stage. PMID- 15109080 TI - Characteristics of behavior associated with alternative choice in a Y maze in rats and their interpretation in terms of complex probability amplitudes. AB - This report demonstrates the possibility of interpreting rat behavior in situations associated with the choice between "safe" locations in a Y maze using pain reinforcement on the basis of summation rules for complex amplitude probabilities characterizing the prognostic assessment by the rat that it will achieve relative safety. Functionally incomplete actions in rat behavior were analyzed in the experimental situation. When the rat needed to avoid pain reinforcement and both pathways to avoid painful stimulation were equally suitable, interference could appear in the animal's assessment of the suitability of the two paths, this preventing decision-making. When this occurred, there was a delay in the executive action. After rats left one of the "safe" arms, they could move into the other "safe" arm, and vice versa. These transfers were completed in a quasi-periodic fashion, as predicted by a formal description of transfers between coincident (or sufficiently similar) states. PMID- 15109081 TI - Blockade of D1 dopamine receptors in the cat motor cortex induces increases in the latent period of a conditioned forepaw support-placing reflex. PMID- 15109082 TI - Pentylenetetrazole kindling induces activation of caspase-3 in the rat brain. PMID- 15109084 TI - Electron microscopic characterization of neurons in the anterior part of the amygdaloid body of the rat brain. PMID- 15109085 TI - Nitric oxide synthase in neurons in the nucleus dorsalis and ganglion nodosum of the vagus nerve and changes during inhalation of acetylcholine in normal conditions and in experimental bronchial asthma. PMID- 15109083 TI - Neurons of layer I and their significance in the embryogenesis of the neocortex. PMID- 15109086 TI - Interaction of sensory neurons and satellite cells during stimulation of nerve regeneration. PMID- 15109087 TI - Functional characteristics of the associative areas of the cortex involved in visual information discrimination learning processes in monkeys. AB - Experiments on three groups of rhesus macaques (intact and with bilateral removal of field 7 and the sulcus principalis) were performed to study the functional characteristics of the associative areas of the cortex while the monkeys learned visual discrimination. Significant differences in learning processes associated with removal of structures and the properties of the stimuli were seen in all animals, in the form of different types of learning curves. As compared with intact monkeys, removal of field 7 had no effect on learning processes for images with properties such as spatial frequency, color, and animal images, though there was a significant worsening in the characteristics of learning during visual discrimination of spatial relationships between objects. Learning processes became unstable, the number of peaks and troughs on learning curves increased, and as a result the training periods were significantly lengthened and 85% of the animals were unable to achieve the learning criterion. Removal of the sulcus principalis significantly worsened the characteristics of discrimination of the sizes of geometric objects, the spatial relationships between them, and stimuli of different colors. The stable reaction time and the probability of refusal in most cases also increased for monkeys of both these groups. Cluster analysis based on the quantitative characteristics of learning processes, despite individual differences between the monkeys, demonstrated a tendency for stimuli to be separated into classes corresponding to different types of information. These data show that the result of sensory processing is that several (at least three) functional visual information flows are generated and that different areas of the cortex deal with these different flows. PMID- 15109088 TI - The correcting influence of the locus ceruleus on ophthalmic hypertension of hypothalamic origin. AB - Chronic experiments were performed on rabbits to study changes in intraocular pressure, the coefficient of flow conductivity, and the chamber fluid minute volume during chronic emotional stress induced by long-lasting repeated electrical stimulation of the ventromedial nucleus of the hypothalamus alone and in combination with electrical stimulation of the locus ceruleus and central gray matter of the periacqueductal matter. Stimulation of the ventromedial nucleus of the hypothalamus was accompanied by an increase in intraocular pressure, an increase in the production of eye chamber fluid, and a decrease in the coefficient of flow conductivity. Electrical stimulation of the locus ceruleus and central gray matter of the periacqueductal matter led to less marked increases in intraocular pressure. Combined stimulation of the hypothalamus and locus ceruleus led to normalization of intraocular pressure due to a reduction in the volume of chamber fluid and an increase in the coefficient of flow conductivity. PMID- 15109089 TI - The effects of transcranial electrostimulation on the adaptive state. AB - The effects of transcranial electrostimulation on medical students with different types of adaptive responses (training, activation, stress, reactivation), levels of reactivity, and psychophysiological and autonomic status were studied. These experiments showed that transcranial electrostimulation was effective for subjects with adaptive responses of the training and activation types but not in those with stress combined with marked vagal tension. Transcranial electrostimulation facilitated improvements in psychophysiological parameters of the students and increased the level of body reactivity. PMID- 15109090 TI - Responses of respiratory neurons in the medulla oblongata to stimulation of the septal nuclei during hypoxia. AB - Dynamic studies were performed in conditions of hypoxia on the effects of stimulation of the ventral, lateral, and medial nuclei of the septum on the spike activity of bulbar respiratory neurons and respiration. The various phases of hypoxia provided a model experiment over which the overall effects of the septal neurons were summed. Electrical stimulation of these septal nuclei in conditions of normal atmospheric pressure had both facilitatory and inhibitory effects on the spike activity of respiratory neurons in the respiratory center of the medulla oblongata, inhibitory effects being predominant. The ventral nucleus had the most effective inhibitory effect on the activity of respiratory neurons. Electrical stimulation of the septal nuclei in the initial phase (4000-5000 m) of hypoxia, on the background of activation, had a predominantly inhibitory influence on the activity of respiratory neurons. During the phase of severe hypoxia (7500-8000 m), on the background of marked hypoxic suppression of respiratory neuron activity, stimulation of the septal nuclei produced no characteristic changes in the activity of these neurons. PMID- 15109091 TI - An outbreak of Cryptosporidium parvum in a Surrey pool with detection in pool water sampling. PMID- 15109092 TI - International note. Global Advisory Committee on Vaccine Safety 11-12 June 2003. PMID- 15109093 TI - Stimuli-sensitive hydrogels: ideal carriers for chronobiology and chronotherapy. AB - The development of solid-phase peptide synthesis in the early 1960s and recombinant DNA technology in the early 1970s boosted the scientific interest of utilizing proteins and peptides as potential therapeutic agents to battle poorly controlled diseases. While there has been rapid progress in the development and synthesis of new proteins and peptides as potential therapeutic agents, the formulation and development of the associated delivery systems is lacking. The development of delivery systems is equally important due to the problems of stability, low bioavailability and short half-life of proteins and peptides. The main problem in this field is that low stability leads to low bioavailability. In this review we draw attention to chrono-pharmacological drug-delivery systems, which can be used to match the delivery of therapeutic agents with the biological rhythm. They are very important especially in endocrinology and in vaccine therapy. We show that the treatment of hypopituitary dwarfism by administration of human growth-hormone-releasing hormone (GHRH) is more effective when GHRH is administered in a pulsatile manner that exhibits a period characteristic of the patient's circadian rhythm. Here we examine how to design novel chrono pharmacological drug-delivery systems that should be able to release the therapeutic agents at predetermined intervals. PMID- 15109094 TI - Drug release from polyureaurethane coating modified by plasma immersion ion implantation. AB - A crosslinked polyurethanurea (PUU) coating was synthesised from a solution on metal vascular stents. In the model system the glucocorticoid prednisolone was inserted into the film by the equilibrium swelling method; after this plasma immersion ion implantation (PIII) was applied to modify the coating for improved release kinetics. This treatment causes the formation of oxygen-containing and unsaturated carbon-carbon groups in the PUU and a destruction of the drug in the surface layer. As a consequence, the release rate of prednisolone to water becomes more stable with time than it is at the untreated coating. In this drug release system PIII treatment prevents an initial toxically high release of the drug. By this it allows the incorporation of a higher amount of the drug and an extended action. PMID- 15109095 TI - Combination of ion beam stabilisation, plasma etching and plasma deposition for the development of tissue engineering micropatterned supports. AB - The performance of biomedical assays at both molecular and cellular level depends greatly on the ability to design new polymer surfaces. Patterns can be created by using materials with contrasted surface properties. In this work we describe in detail the preparation of micropatterned surfaces to be used as tissue engineering supports. Poly(ethylene glycol) (PEG) was used as the 'anti-fouling' polymer in opposition to functional regions covered by acrylic acid (AAc). Since spin-casted PEG films are unstable, ion beam stabilization (IBS) treatment was applied in order to render it insoluble. On the other hand, AAc films were deposited by low-power plasma chemical vapour deposition. Chemical properties of both polymers were monitored by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy while topographic features were followed by atomic force microscopy. Finally, a micropattern was produced by using a mask, which isolated the IBS-PEG from the AAc-deposited regions. Endothelial cells cultured on the surface were observed to follow the micropatterns. In fact, for a certain surface density it was observed that the cells present tensile or compressive stresses when forced to remain in the anti-fouling or the functionalised regions, respectively. PMID- 15109096 TI - Biocompatibility of platinum-metallized silicone rubber: in vivo and in vitro evaluation. AB - Silicone rubber is commonly used for biomedical applications, including implanted cuff electrodes for both recording and stimulation of peripheral nerves. This study was undertaken to evaluate the consequences of a new platinum metallization method on the biocompatibility of silicone rubber cuff electrodes. This method was introduced in order to allow the manufacture of spiral nerve cuff electrodes with a large number of contacts. The metallization process, implying silicone coating with poly(methyl methacrylate) (PMMA), its activation by an excimer laser and subsequent electroless metal deposition, led to a new surface microtexture. The neutral red cytotoxicity assay procedure was first applied in vitro on BALB/c 3T3 fibroblasts in order to analyze the cellular response elicited by the studied material. An in vivo assay was then performed to investigate the tissue reaction after chronic subcutaneous implantation of the metallized material. Results demonstrate that silicone rubber biocompatibility is not altered by the new platinum metallization method. PMID- 15109097 TI - Polymer composition and acidification effects on the swelling and mechanical properties of poly(acrylamide-co-acrylic acid) superporous hydrogels. AB - Poly(acrylamide-co-acrylic acid) (poly(AM-co-AA)) superporous hydrogels (SPHs) were synthesized and the acidification effects on the swelling and mechanical properties were studied. Gelation exotherms were measured to determine the optimum introduction time for adding a blowing agent. The gelation kinetics decreased with increasing the AA concentration. The maximum equilibrium swelling was observed around an AA weight fraction of 0.4, but the compressive strength decreased monotonically with increasing the AA concentration. Poly(AM-co-AA) SPHs were much less swollen in acidic solution than in distilled water of pH 6.7. The swelling ratio decreased with increasing acidity (decreasing pH). Reduction of water absorption content by acidification led to considerable increase in the mechanical strength. PMID- 15109098 TI - Biocompatibility and biodegradation of cross-linked gelatin/hyaluronic acid sponge in rat subcutaneous tissue. AB - A gelatin/hyaluronic acid (GH) sponge has been fabricated by freeze-drying and cross-linking. The GH sponge was insoluble when cross-linked with 1-ethyl-3-(3 dimethylaminopropyl) carbodiimide. The morphologies of sponges were investigated using a field emission scanning electron microscope. The porosity of the GH sponge increased with hyaluronic acid content. The GH sponge was biodegradable, as evidenced by implantation in Wistar rat subcutaneous connective tissue. Fibroblasts infiltrated into the sponge matrix, and regenerated collagen in the matrix to a level of 25% by 15 days after surgery. The GH73 sponge induced an acute inflammatory response compared with the GH91 sponge. This inflammatory response could have been stimulated by the presence of hyaluronic acid up to Day 10, as it decreased afterwards. The C-reactive protein of blood samples also indicated the same result. The blood tests and histological results show that GH sponges have good biocompatibility and low antigenicity for tissue engineering scaffolds. PMID- 15109099 TI - Controlling modulus and morphology of hydrogel tubes through surface modification. AB - Crosslinked, porous poly(2-hydroxyethyl methacrylate-co-methyl methacrylate) (PHEMA-MMA) tubes were prepared in cylindrical glass molds using a new centrifugal casting process developed in our group. The resulting hydrogel tubes have a bi-phasic wall structure, with a spongy inner layer and a gel-like outer layer, the latter of which provides mechanical strength to the tube. While many factors influence wall morphology and, thus, mechanical properties, we focused on the effect of the surface properties of the glass mold in which tubes are synthesized. Specifically, we investigated the impact of a diverse set of silane modifications of the glass mold on tube morphology, elastic modulus and mold release. We treated activated glass surfaces with one of three alkoxysilanes having either ethoxy, amine or fluorocarbon end-groups. Silane-modified glass surfaces were found to be more hydrophobic than the unmodified glass mold, with the most hydrophobic surface being that of the fluorocarbon-terminated silane. The presence of the silane layer on the mold was confirmed by X-ray photoelectron spectroscopy and the stability of this modification was confirmed by examining the surface chemistry of the hydrogel tubes. The biphasic hydrogel tube wall structure was observed for all tubes, yet those tubes synthesized in unmodified molds had a cracked outer morphology, whereas those synthesized in silane modified molds had a smooth outer morphology. This influenced the mechanical properties of the tubes where tubes synthesized in silane-modified molds had a significantly greater elastic modulus than those tubes synthesized in unmodified molds. Release from the molds was easiest with ethoxy- and amine-functionalized silane mold modifications. PMID- 15109100 TI - Rhamnolipid biosurfactant behavior in solutions. AB - The solution behavior of rhamnolipid biosurfactant that is determined by its surface thermodynamic properties often plays a central role in determining and controlling its performance in practical applications. In this study, rhamnolipid molecules exhibited a predominant hydrophilic surface, contributed by hydrophilic monomer head groups. Interaction free energies between rhamnolipid molecules at water surfaces, as estimated based on the distance-dependent decaying interaction contact areas, decayed exponentially with regards to separation distance. These interaction free energies were governed by hydrophilic monomer head groups of rhamnolipid molecules. The experimental observations in this study were consistent with the previous postulations. PMID- 15109101 TI - Salmon fibrin supports an increased number of sprouts and decreased degradation while maintaining sprout length relative to human fibrin in an in vitro angiogenesis model. AB - Salmon-derived fibrin has been proposed as a preferred alternative to human or bovine fibrin because of its reduced potential for disease transmission. Here we evaluate salmon fibrin as an alternative ECM support for therapeutic angiogenesis applications, such as vascularizing engineered tissues. Human umbilical vein endothelial cells (HUVEC) seeded on gelatin beads and suspended in either salmon or human fibrin sprouted and formed capillary-like structures. Sprout length was generally increased with the addition of bFGF and VEGF and further increased with the addition of phorbol myristate acetate (PMA). The number of sprouts per bead was increased 61-188% in salmon fibrin relative to human fibrin (alpha < 0.0005) in cultures receiving growth factors and PMA, while average sprout lengths were similar for HUVEC within human or salmon fibrin. Additionally, under these conditions in the absence of a protease inhibitor, HUVEC appeared to degrade human, but not salmon, fibrin. These results support the idea that salmon fibrin may be an attractive alternative ECM able to support microvascular network formation. PMID- 15109102 TI - [The contribution of neuroimaging for speech diagnosis and therapy]. PMID- 15109103 TI - [Facial measurements and proportions in children: contributions to orofacial myofunctional evaluation]. AB - BACKGROUND: facial measurements and proportions are very important for the orofacial myofunctional evaluation. AIM: to describe the facial measurements and proportions, according to age and gender. METHOD: 254 children, with ages ranging between 7.0 to 11.11 years. RESULTS: there was no statistical difference between the mean values, regarding age and gender, for the upper lip and philtrum. As for the other facial measurements a statistically significant difference was observed between the obtained mean values. Girls presented lower mean values than boys. The facial proportions did not present statistical difference between the mean values in relation to age and gender. CONCLUSION: it is important to consider aspects of age and gender during the evaluation of the facial measurements. PMID- 15109104 TI - [Lisping and dental occlusion alterations in 03 to 06 year old children]. AB - BACKGROUND: lisping and dental occlusion alterations. AIM: to verify the relationship between lisping and alterations in the anterior vertical plane of the dental occlusion in children with ages ranging from 03 to 06 years. METHOD: 132 children were submitted to an evaluation of the dental occlusion and of speech. RESULTS: lisping occurred in 51.5% of the participants and was associated to dental occlusion alterations in 50% of the individuals, while for the other 50% lisping was not associated to any dental occlusion alterations. CONCLUSION: lisping did not demonstrate to have any association with alterations in the anterior vertical plane of the dental occlusion. The presence of alterations in the dental occlusion can be a risk factor for the development of lisping in speech, but is not decisive for its occurrence. PMID- 15109105 TI - Acoustic analyses of speech naturalness: a comparison between two therapeutic approaches. AB - BACKGROUND: acoustic analyses of speech naturalness. AIM: to use voice onset time (VOT) measure as a parameter of speech naturalness, comparing the outcome of two stuttering treatment procedures. METHOD: 12 adult stutterers were divided in two groups and submitted to the same Fluency Promotion Program (FPF). For GI: surface electromyography (EMG) was used as biofeedback. RESULTS: the results indicate that for GI 50% of the participants presented a more natural sounding speech after the FPF, whereas for GII only 17% of the participants demonstrated adequate speech naturalness in the same period. CONCLUSION: although the number of participants of this research was small, the association of EMG to a FPP seems to produce a more natural sounding speech. PMID- 15109106 TI - [Dysarthria: a speech rate study]. AB - BACKGROUND: speech rate in dysarthric patients. AIM: to obtain an objective measure of the speech rate of dysarthric patients during the reading of a standardized text and during spontaneous speech. METHOD: eleven subjects, with ages ranging from 18 to 69 years, were studied. For each dysarthric patient, 3 normal individuals of the same gender and age were evaluated, composing the control group. The speech samples were collected based on a reading of a standardized text and on a two minute conversation. Speech rate was analyzed in terms of words per minute. RESULTS: the analysis of the results demonstrated that in both situations of speech, the dysarthric group presented a statistically significant slower speech rate than the control group. CONCLUSION: the values of speech rate obtained in this study do not agree with other studies carried out in the English language. PMID- 15109107 TI - [Picture naming and memory in children: phonological and semantic effects]. AB - BACKGROUND: [corrected] The relation between picture naming and the short and long term memories. AIM: to verify the ability of picture naming based on phonological and semantic queues, relating it to memory. METHOD: 80 pictures selected from a set of 400 (Cycowicz et al., 1997) were presented to 80 children with ages ranging from 3 to 6 years. Responses were classified in semantic and phonologic errors and number of correct answers. RESULTS: The effect of the articulatory complexity was significant and the effect of the semantic complexity was not significant. CONCLUSION: Naming is the result of memory activation which is organized in categories, physical properties and function; phonologic effects do interfere in the activity of naming, whereas the semantic effects reflect that the long term memory is organized in categories which are dependant of the context and of the development. PMID- 15109108 TI - [Pragmatic abilities in the discourse of children with and without specific language impairment]. AB - BACKGROUND: to investigate the pragmatic difficulties presented by children with specific language impairment and to verify how these deficits influence their discourse performance. AIM: to analyze the request-response sequences in the discourse of normal and Specific Language Impairment Children (SLI) during a 20 minute free-play situation with an adult. METHOD: subjects of this study were 29 children, male and female, with ages ranging from 2:0 to 4:11 years. Data was collected during a free-play situation with an adult. RESULTS: SLI children frequently ignored the questions made by the adult and responded innappropriately more often. CONCLUSION: qualitative and quantitative differences in the pattern of responses and in the use of pragmatic abilities were found for the SLI children. PMID- 15109109 TI - [The communication of autistic spectrum children in group activities]. AB - BACKGROUND: language is the most useful diagnostic feature in cases of infantile autism, and is many times connected with the prognosis. OBJECTIVE: to identify the communication profile of autistic spectrum children through the analysis of the functional use of language in two different communication situations. METHOD: five groups of recordings of 10 institutionalized children, during a period of 12 months, in two different communication situations: situation I (in a group with co-ordination) and situation II (in a group without co-ordination). RESULTS: in both situations of communication there was a significant change in the communication profile of these children. CONCLUSION: it can be observed that these children do perceive the differences present in situations of communication. PMID- 15109110 TI - [Working memory in children with phonological deviation]. AB - BACKGROUND: working memory. AIM: to verify the performance of children with normal speech development (NSD) and with evolutional phonological deviation (EPD), in tasks which demand the use of working memory. METHOD: as a research instrument, subtest 5 of the Illinois Test of Psycholinguistics Abilities (ITPA) was used, as well as the meaningless word repetition test. The answers were analyzed based on the results of the digit repetition test and the meaningless word repetition test, comparing the performance of both groups. RESULTS: when comparing the groups, it was verified that the performance of children with EPD is lower than children with NSD in both of the analyzed tests. CONCLUSION: the obtainded results confirm that the working memory is fundamental for the development of speech and for the phoneme choice when producing words. PMID- 15109111 TI - [A study of hearing and of the auditory habits of youngsters of the city of Sorocaba--Sao Paulo]. AB - BACKGROUND: currently the exposure to noisy leisure activities are part of the routine of youngsters, representing a considerable risk for hearing. AIM: to study the hearing and the auditory habits of youngsters. METHOD: 75 individuals, with ages ranging from 18 to 25 years, were evaluated using pure tone audiometry and a questionnaire for the verification of their auditory habits. RESULTS: the audiometric results indicate that 45.3% of the individuals presented some sort of hearing impairment, in the 3 to 6 kHz frequency range. As for the auditory habits, most of the youngsters have the habit of exposing herself/himself to high sound pressure levels, especially collective music. CONCLUSION: It is essential for the health professionals to be enrolled in prevention and hearing conservation programs in order to change attitudes and to establish noise control measures for leisure activities. PMID- 15109112 TI - [Ear canal narrowing: a case report]. AB - BACKGROUND: ear canal narrowing is the thinning or closing of the ear canal. This can damage the sound transmition from the external to the internal ear, which in turn may cause a false hearing loss. AIM: to alert the professionals of the area on how to avoid a miss diagnosis. METHOD: a 47 year old woman, submitted to two audiologic evaluations: one without avoiding ear canal closing and one avoiding it. RESULTS: a decrease of up to 40 dB was observed in the results of the second evaluation. CONCLUSION: in order to obtain reliable results it is extremely important to verify the presence of ear canal narrowing, at any age, and to use appropriate procedures to avoid it from happening during testing. PMID- 15109113 TI - [The use of the self-assessment questionnaire to evaluate hearing aid benefit]. AB - BACKGROUND: assessment of the subjective benefit brought by the use of the hearing aid. AIM: to determine the benefit brought by the use of the hearing aid through a subjective procedure. METHOD: the subjective benefit of the hearing aid was assessed in 15 hearing impaired individuals, novice hearing aid users, through the Abbreviated Profile of Hearing Aind Benefit (APHAB) self-assessment questionnaire, which was answered before and after the hearing aid fitting. RESULTS: the results demonstrated statistically significant differences between the conditions with and without the hearing aid, indicating a better performance with the use of amplification. CONCLUSION: this procedure demonstrated to be valuable for the assessment of the benefits brought by the process of hearing aid fitting. PMID- 15109114 TI - [Communication strategies used by individuals with moderate sensorineural hearing loss]. AB - BACKGROUND: communication strategies (CS) are mechanisms used to facilitate speech comprehension and the transmission of a message to another speaker. AIM: to characterize the communication strategies used by 20 adults, fitted with hearing aids, with moderate sensorineural hearing loss. METHOD: a 60 minute interview, with an audiologist, with and without the hearing aid. RESULTS: the CS which prevailed were those of a cognitive and palliative nature in the situations with and without the hearing aid. CONCLUSION: most of the individuals used the communication strategies to facilitate the recognition of the message. PMID- 15109115 TI - [Brazilian Ph.D. speech-language pathologist and audiologists: analysis of thesis material according to the field of practice and programs]. AB - BACKGROUND: Ph.D. programs in Speech-Language and Hearing Science. AIM: to bring to date the concluded doctorate thesis of Brazilian speech-language and hearing pathologists, until December of 2003. CONCLUSION: from the profile analysis of the concluded doctorate thesis of Brazilian speech-language pathologists, it is possible to conclude that in the period between 1976, when the first thesis was concluded, to December of 2003, a total of 203 thesis elaborated by Ph.D. speech language and hearing pathologists were found (in its majority women), in the areas of Hearing, Language, Voice and Oralfacial Myofunctional system. PMID- 15109116 TI - Physical aggression, forced sex, and stalking victimization by a dating partner: an analysis of the National Violence Against Women Survey. AB - This study used the National Violence Against Women Survey (NVAWS) of women and men to estimate noncohabitating dating violence prevalence by type (physical, forced sex, and stalking), associations between dating violence and other types of interpersonal violence across the lifespan, and association of dating violence with longer-term mental health including substance abuse. Among respondents aged 18 to 65, 8.3% of 6,790 women and 2.4% of 7,122 men experienced physical aggression, forced sex, or stalking victimization by a dating partner. Few (20.6% of women and 9.7% of men) reported more than one type of dating violence. Childhood physical aggression by a parent or guardian was strongly associated with subsequent dating violence risk for men and women. Dating violence (physical aggression specifically) was associated with current depressive symptoms, current therapeutic drug use (antidepressants, tranquilizers, or pain medications), and current recreation drug use for women. Implications for parents, survivors, health care, and service providers are discussed. PMID- 15109117 TI - The joint contribution of experiencing and witnessing violence during childhood on child abuse in the parent role. AB - This article examines adult respondents' abuse of children as a consequence of their own childhood experiences of abuse, both direct experiences of childhood violence (hitting) and exposure to interparental violence (witnessing). In particular, the study examines the extent to which these factors function interactively: Are both experience and exposure necessary or is either sufficient to increase disproportionately the probability of child abuse? Using data from the Second National Family Violence Survey, results of a logistic regression analysis show that either or both factors produced higher than average and relatively similar rates of child abuse. Only respondents with neither form of family violence reported lower than average rates of abuse of their own children. The analysis controlled for gender, race, family income, and family structure; race was the only control variable to be significantly associated with child abuse. Finally, no control variable modified the interaction between the family violence variables. PMID- 15109118 TI - The internal validity of the index of spouse abuse in African American women. AB - This study investigated the internal validity of the Index of Spouse Abuse (ISA) (Hudson & McIntosh, 1981) in a sample of 583 African American women who sought health care at a tertiary care hospital in Atlanta, Georgia. Three models were tested with confirmatory factor techniques: (a) Hudson and McIntosh's original Index of Spouse Abuse two factor model; (b) Campbell, Campbell, Parker, and Ryan's three factor model (Campbell, Campbell, King, Parker, & Ryan, 1994); and (c) an alternative model of physical and nonphysical abuse. This alternative model is based in part on Tolman's conceptualization of psychological abuse as a construct comprised of two related but distinct factors: controlling and emotionally abusive behaviors (Tolman, 1999). Results show that the alternative model fits the data better than the first two models. Findings support the continued use of the ISA, but with proposed modifications. PMID- 15109119 TI - Perceptions of domestic abuse in same-sex relationships and implications for criminal justice and mental health responses. AB - Male and female university students (N = 171) read scenarios describing a domestic abuse incident that systematically varied the sex of victim and perpetrator to produce four between-participants conditions. Results were consistent with and extended previous findings about perceptions of heterosexual domestic abuse to include gay and lesbian domestic abuse. Findings suggest that participant perceptions of abuse in same-sex and heterosexual relationships are similar. Where they differ, the differences have significant implications: (a) participants considered male against female abuse to be more serious than same sex domestic abuse, (b) participants were more likely to recommend that the victim press charges in male against female abuse than in same-sex domestic abuse, (c) participants perceived same-sex victims to be less believable than heterosexual victims, and (d) victim believability was correlated with sentencing recommendations. Implications for criminal justice and mental health intervention are considered. Directions for future research are outlined. PMID- 15109120 TI - An analysis of the self-control and criminal versatility of gang and dating violence offenders. AB - How versatile are gang and dating violence offenders? Current gang research highlights the versatility of gang members, yet the versatility of intimate violence offenders is often unexamined. Gottfredson and Hirschi, A General Theory of Crime (1990), support the idea of versatile rather than specialized offenders and suggests that low self-control is associated with a host of criminal and noncriminal risk-taking activities. Using data from a self-report sample of 1139 youths in grades 9 through 11, we investigated both the versatility of gang and dating violence offenders and theoretical variables associated with each. We find disproportionate offending by dating and gang violence offenders in a variety of crimes, as well as considerable overlap in the independent variables associated with both types of violence. Low levels of self-control and exposure to general and crime-specific criminal opportunities are significantly associated with engaging in dating and gang violence. PMID- 15109121 TI - Exposure to "low-level" aggression in school: associations with aggressive behavior, future expectations, and perceived safety. AB - Examined associations with witnessing and being victimized by "low-level" aggressive acts (e.g., pushing, gossip) and three indicators of psychosocial functioning in a sample of 771 elementary school students from one urban and one suburban school district. Results indicated that exposure to low-level aggression appears to relate to psychosocial functioning in ways similar to more severe forms of aggression. Students who were exposed to higher levels of both witnessing and victimization by low-level aggression reported the highest levels of engagement in aggression, the lowest levels of positive expectations for the future, and the lowest levels of perceived safety. Findings are discussed in the context of research on exposure to aggression in general, with suggestions offered for future studies. Implications of the findings for school-based intervention programs are raised. PMID- 15109122 TI - Serotypes of Haemophilus influenzae strains isolated from pediatric patients with respiratory tract infections. AB - Capsular serotypes of 122 respiratory Haemophilus influenzae strains were identified utilizing antigenic examinations (Slide Agglutination Test [SAT] and counter immuno-electrophoresis [CIE]), and PCR method. Results of the two methods were compared to find disagreement. Clinical and bacteriological characteristics of encapsulated strains were analyzed. By utilizing PCR assay, 3 (2.5%) serotype b, 1 (0.8%) serotype f and 118 (96.7%) non-typeable strains were found among 122 tested strains. Discrepancy between the results of antigenic examinations and PCR assay was found in one strain. That strain gave positive results by antigenic tests with capsular type c antiserum but it was determined to be capsular type f by PCR. No serotype b- strain was detected. All the encapsulated strains displayed biotype I. All the serotype b strains produced beta-lactamase, but none of those was sulbactam/ampicillin and amoxicillin/clavulanic acid resistant. The serotype f strain was beta-lactamase non-producing ampicillin susceptible. The patients suffered from respiratory infections due to encapsulated strains displayed safe clinical manifestations and good clinical responses to antibiotic treatments using beta-lactams. In pre-vaccination era, PCR is considered to be a useful method for determination of serotypes of respiratory H. influenzae, which may contribute to reducing a possibility of clonal transmission of serotype b strains among children community, which is one of the potential risk factors for pediatric invasive infections. PMID- 15109123 TI - Effect of dose-rate and lung dose in total body irradiation on interstitial pneumonitis after bone marrow transplantation. AB - The aim of this study is to evaluate the incidence of interstitial pneumonitis following fractionated total body irradiation conditioning for bone marrow transplantation with varying lung doses due to shielding technique and different dose-rates. Between 1987 and 2001, a total number of 105 patients have received total body irradiation conditioning for bone marrow transplantation for hematological malignancies at Gulhane Military Medical School. Twelve Gy fractionated total body irradiation was delivered in 6 fractions over 3 consecutive days with Co-60 teletherapy machine. Conditioning therapy included only cyclophosphamide (60 mg/ kg/day for two days) and total body irradiation. The median follow-up for patients was 12 months. Interstitial pneumonitis developed in 10 patients out of 105 patients (9.52%). The median total dose to lung was 9.60 Gy (8.88-10.90). The difference between total lung dose and interstitial pneumonitis was not significant. Pneumonitis development in the high dose-rate (>0.04 Gy/min) group versus low dose-rate (< or =0.04 Gy/min) group was statistically significant. Low dose-rate fractionated total body irradiation is a reliable conditioning program in bone marrow transplantation with effective lung sparing to avoid interstitial pneumonitis. PMID- 15109124 TI - Assessment of in-hospital cardiopulmonary resuscitation using Utstein template in a university hospital. AB - The aim of this study was to evaluate the effectiveness of in-hospital cardiopulmonary resuscitation (CPR) strategies and identify key predictors of post-CPR survival in a university hospital setting. Using a form recommended by the European Resuscitation Council, data regarding in-hospital CPR attempts from January 2001 to December 2002 were recorded and analyzed. The main outcomes of interest were immediate survival after CPR and survival to hospital discharge. Of 307 patients who suffered cardiac arrest in the study period, 103 (33.5%) were resuscitated. Of these 103 patients, 28 (27.2%) survived immediately and 12 (11.7%) survived to hospital discharge. The key predictors of immediate survival were CPR duration and initial cardiac rhythm as monitored by ventricular fibrillation/pulseless ventricular tachycardia (VF/VT). The key predictors of survival to hospital discharge were CPR duration, immediate defibrillation, Glasgow Coma Scale score, and Early Prediction Score. Together, our results suggest that in-hospital CPR strategies require improvement. They also underscore the importance of data collection and analysis in evaluating the effectiveness of inhospital CPR strategies. PMID- 15109125 TI - Long-term use of corticosteroid eye drops delays the spontaneous remission of pulmonary sarcoidosis. AB - Topical corticosteroid eye drops are commonly used for ocular sarcoidosis. That systemic absorption of corticosteroids by eye drops may influence the clinical course of sarcoidosis may be speculated because it has been reported that the serum concentration of corticosteroids after drop administration was dose related. To evaluate the effects of corticosteroid eye drops on the clinical course of patients with stage I pulmonary sarcoidosis, we compared the serum levels of angiotensin converting enzyme (ACE) and bilateral hilar lymphadenopathy (BHL) on chest radiographs of group CS, which is consisted of patients who received topical therapy of betamethasone in the form of eye drops for anterior uveitis, and group CN, which is consisted of patients who did not receive any medications throughout the entire course of the disease. Although the serum ACE level was not significantly different between groups CS and CN at the time of the diagnosis of pulmonary sarcoidosis, the level of serum ACE in group CS was significantly higher than that in group CN 20 months after the topical corticosteroid treatment (24 IU/ml and 16 IU/ml, respectively). Further, the size of BHL on chest radiography in group CS was significantly larger than that in group CN 20 months after the topical treatment (82% and 37% of before control, respectively). These findings suggest the possibility that the topical corticosteroid therapy influenced the clinical course of pulmonary sarcoidosis, inducing some delay in the spontaneous remission in the longterm course. PMID- 15109126 TI - The effects of vagal stimulation on laryngeal vascular resistance and intraluminal pressure in the dog. AB - In anaesthetized dogs (sodium pentobarbitone 30 mg/kg, i.v.) laryngeal vascular resistance was measured by unilateral perfusion at constant flow of the branch of the cranial superior thyroid artery that supplies the larynx. Arterial perfusion was at constant flow and inflow pressure was divided by flow to give laryngeal vascular resistance (R(LV)). Intraluminal laryngeal pressure (P(L)) and systemic arterial blood pressure (BP) were also measured. Stimulation (20 V, 20 Hz, 0.2 milliseconds) of the central end of cervical vagus caused an increase in R(LV) (+22.9+/-6.1%) and a decrease in P(L) (-12.1+/-4.4%). Stimulation (10 V, 10 Hz, 0.2 milliseconds) of the central end of the recurrent laryngeal nerve (RLN) reduced RLV (-3.4+/-0.8%) and P(L) (-7.5+/-4.1%). Stimulation of the peripheral end of the RLN decreased R(LV) (-7.1+/-1.9%) and increased PL (+21.6+/-7.7%). Stimulation of the central end of the superior laryngeal nerve (SLN) increased R(LV) (+17.9+/-3.2%) and P(L) (+59.8+/-2.7%), whereas stimulation of the peripheral end of the SLN decreased R(LV) (-4.8+/-1.6%) and P(L) (-4.1+/-2.4%). After treatment with alpha-adrenoreceptor antagonist phentolamine (0.5 mg/kg, i.v.), stimulation of the central end of cervical vagus nerve reduced R(LV) by 25% and decreased BP. Phentolamine caused a decrease in BP and reduced the magnitude of increase in R(LV) in response to stimulation of central end of SLN. After atropine sulphate (0.5-2.0 mg/kg, i.v.), the stimulation of both central and peripheral ends of RLN reduced R(LV). The decrease in R(LV) during stimulation of peripheral end of SLN was reduced by atropine. Thereafter, pancuronium bromide (0.06-0.1 mg/kg, i.v.) was given and dogs were artifically ventilated. After paralyzed, stimulation of the central end of the SLN decreased R(LV) (+26.0+/-4.5%) but produced no change in P(L), It is concluded that parasympathetic motor fibers in the RLN and SLN are effective for the laryngeal vascularity and non-adrenergic system may be responsible for laryngeal vasoconstriction. laryngeal vasculature; vagal stimulation; phentolamine; atropine PMID- 15109127 TI - Pneumocele vs. pneumosinus dilatans: review of the literature with a case of frontal sinus pneumocele. AB - Pneumoceles of the frontal sinus are very rare conditions, characterized by abnormal expansion of the frontal sinus. The patient usually complains of slowly changing facial contours. The etiology and pathogenesis of pneumoceles remains unclear. A case of pneumocele of the frontal sinus is presented, and the differentiated features of the disease are discussed. PMID- 15109128 TI - Panniculitis of the descending colon caused by enterocolic phlebitis: a case report. AB - A 73-year-old male was referred to our hospital for abdominal pain, diarrhea and general fatigue lasting for 3 weeks. Physical examination of the abdomen revealed a firm mass in the left abdominal region. Computed tomography revealed a mass around the descending colon. Colonoscopy and barium enema revealed poor extensibility of the lumen with edematous mucosa, and narrowing of the descending colon with rugged mucosal surface. Because of the clinical symptoms and findings, the patient was diagnosed clinically as suffering from panniculitis of the descending colon. He underwent the left hemi-colectomy with side-to-side colo colostomy after making of a loop ileostomy. Histological analysis of the resected colon showed an infiltration of inflammatory cells, predominantly lymphocytes, into veins and venules of the submucosa, muscularis propria and fat tissue of the colonic mesentery, with an involvement of all layers of the vessel wall. Arteries were escaped from inflammatory changes. The histopathological diagnosis of enterocolic phlebitis and venulitis was made because of these findings. PMID- 15109129 TI - Villoglandular papillary adenocarcinoma of the uterine cervix in a pregnant woman: a case report and review of literature. AB - Villoglandular papillary adenocarcinoma (VPA) of cervix is rare but a well recognized variant of cervical adenocarcinoma with favorable prognosis occurring in younger age group. A 28-year-old white woman, gravida 3, para 2 was admitted for abnormal vaginal bleeding, when she was pregnant at 8th weeks of gestation. Physical examination revealed about 2.5 cm polipoid lesion of the cervix protruding into vagina. Histopathological findings were consistent with cervical VPA. After termination of pregnancy, radical hysterectomy type III was performed. The patient underwent second, third and fourth laparotomies because of recurrent pelvic masses. At the end of five years follow-up period, she died because of the complication of recurrent tumor. VPA is not an innocent tumor, and can be complicated by recurrence and metastasis. More radical surgical and medical attempts should be planned. PMID- 15109130 TI - Pathology and ultrastructure of an intranuclear bacilliform virus (IBV) infecting brown shrimp Crangon crangon (Decapoda: Crangonidae). AB - The brown shrimp Crangon crangon supports an important fishery in Europe (over 25000 t, valued at 80 million euros in 2000). Through the course of histopathological screening of crustaceans from the Clyde estuary, western Scotland, for the biological effect of contaminants, we have discovered a highly prevalent (up to 100%) non-occluded intranuclear bacilliform virus (IBV) infection in the hepatopancreatic tubule epithelia and midgut epithelia of wild C. crangon. This is the first report of an IBV in this family. We have termed this virus Crangon crangon bacilliform virus (CcBV). Histological and ultrastructural observations suggest that this virus is similar to other IBVs previously described from crabs and penaeid shrimps. The nuclei of virus-infected epithelial cells contained an eosinophilic, hypertrophied viroplasm that marginalised the chromatin of the host nucleus. Infected cells were often separated from their neighbouring cells and their nuclei appeared apoptotic. In heavily infected shrimp, apoptotic cells were expelled into the lumen of the hepatopancreatic tubule or the midgut. Following this stage, some hepatopancreatic tubules became degenerate, with remnants of the basement membrane and myoepithelial lining remaining. Transmission electron microscopy of hypertrophic nuclei revealed the presence of rod-shaped and cylindrical, envelope bound virions. These virions did not form arrays and were not embedded within occlusion bodies, but did appear to be partially occluded in an amorphous matrix that corresponded to a granular viroplasm. The ultrastructure, morphology and size of the nucleocapsid and the complete virion aligns the virus most closely to the IBVs previously reported from other decapod crustaceans. Due to the pathological manifestation of IBV infection in C. crangon, it appears likely that it can act as a population modulator, particularly at sites where infection prevalence is high, such as that observed in the Clyde estuary. PMID- 15109131 TI - Experimental infection of rainbow trout Oncorhynchus mykiss with viral haemorrhagic septicaemia virus isolates from European marine and farmed fishes. AB - The susceptibility of rainbow trout Oncorhynchus mykiss to infection with various isolates of viral haemorrhagic septicaemia virus (VHSV) was examined. A total of 8 experiments with rainbow trout ranging from 0.6 to 6.2 g was conducted for 139 isolates originating from wild marine fishes in European waters (115 isolates), farmed turbot from Scotland and Ireland (2 isolates), and farmed rainbow trout (22 isolates). The isolates were tested by immersion and/or intraperitoneal injection either as pooled or single isolates. The isolates from wild marine fishes did not cause mortality by immersion while some of the isolates caused mortality when injected. All VHSV isolates from farmed rainbow trout caused significant mortality by immersion. Currently, pathogenicity trials are the only way to differentiate VHSV isolates from wild marine fishes and farmed rainbow trout. The 2 farmed turbot isolates did not cause mortality by immersion, supporting the view that they originated from the marine environment. PMID- 15109132 TI - Experimental horizontal transmission of viral hemorrhagic septicemia virus (VHSV) in Japanese flounder Paralichthys olivaceus. AB - Infection by viral hemorrhagic septicemia virus (VHSV) has recently occurred among wild and farmed Japanese flounder Paralichthys olivaceus in Japan. In the present study, horizontal transmission of VHSV among Japanese flounder was experimentally demonstrated by immersion challenge. Exposure to a flounder isolate (Obama25) of VHSV revealed a dose-response, with higher mortality (81 and 70%) at the 2 higher exposure levels (6.0 and 4.0 log10 TCID50 ml(-1)). In a second experiment, high titers of VHSV were expressed from moribund and dead flounder based on virus detection in holding-tank waters 2 to 3 d prior to death of the fish and 1 d after death. The virus could not be detected in tank waters 2 d after death. Finally, a third cohabitation experiment in small tanks demonstrated horizontal transmission of VHSV from experimentally infected to uninfected fish. PMID- 15109133 TI - Complete sequence of RNA1 and subgenomic RNA3 of Atlantic halibut nodavirus (AHNV). AB - The Nodaviridae are divided into the alphanodavirus genus, which infects insects, and the betanodavirus genus, which infects fishes. Betanodaviruses are the causative agent of viral encephalopathy and retinopathy (VER) in a number of cultivated marine fish species. The Nodaviridae are small non-enveloped RNA viruses that contain a genome consisting of 2 single-stranded positivesense RNA segments: RNA1 (3.1 kb), which encodes the viral part of the RNA-dependent RNA polymerase (RdRp); and RNA2 (1.4 kb), which encodes the capsid protein. In addition to RNA1 and RNA2, a subgenomic transcript of RNA1, RNA3, is present in infected cells. We have cloned and sequenced RNA1 from the Atlantic halibut Hippoglossus hippoglossus nodavirus (AHNV), and for the first time, the sequence of a betanodaviral subgenomic RNA3 has been determined. AHNV RNA1 was 3100 nucleotides in length and contained a main open reading frame encoding a polypeptide of 981 amino acids. Conservative motifs for RdRp were found in the deduced amino acid sequence. RNA3 was 371 nucleotides in length, and contained an open reading frame encoding a peptide of 75 amino acids corresponding to a hypothetical B2 protein, although sequence alignments with the alphanodavirus B2 proteins showed only marginal similarities. AHNV RNA replication in the fish cell line SSN-1 (derived from striped snakehead) was analysed by Northern blot analysis, which indicated that RNA3 was synthesised in large amounts (compared to RNA1) at an early point in time post-infection. PMID- 15109134 TI - Histological, ultrastructural, and in situ hybridization study on enlarged cells in grouper Epinephelus hybrids infected by grouper iridovirus in Taiwan (TGIV). AB - Grouper iridovirus in Taiwan (TGIV) infection in the Epinephelus hybrid is a major problem in the grouper industry. ATPase gene sequences indicate that this virus is closely related to cell hypertrophy iridoviruses. Histologically, the appearance of basophilic or eosinophilic enlarged cells in internal organs is the most characteristic feature of this disease. These cells are acid-phosphatase positive and are able to phagocytose injected carbon particles. In our study, TGIV infection inhibited normal phagocytic ability in these cells in vivo after 4 d post-infection (p.i.) but not before 2 d p.i. Their staining properties and phagocytic ability suggested a monocyte origin of enlarged cells, which appeared in high numbers in the trunk kidney, head kidney, spleen and gill. After infection, the enlarged cells first appeared in the spleen, with an abundance peak at 64 h p.i. (Peak 1); at 120 h p.i., a second peak (Peak 2) occurred in the spleen, head kidney, trunk kidney and gill. Lower numbers of enlarged cells were observed in the liver, muscle, heart, eye, intestine, but no enlarged cells were found in the brain. A TGIV-specific DNA probe labeled most of the basophilic but not eosinophilic enlarged cells. Nuclei of infected cells were labeled during an early stage of the infection; at later stages, both nuclei and cytoplasms were labeled. Ultrastructurally, heterochromatins of the infected cells were marginated or aggregated to one side of the nuclei during the early stages of infection. Damage and rupture of the nuclear membrane started before formation of the viromatrix. Capsids were assembled in ring-shaped or disc-shaped structures. Bullet-shaped electron-dense material was present near the incomplete virus particles, and is speculated to be inserted into the capsids later. PMID- 15109135 TI - Phylogenetic study and identification of Vibrio splendidus-related strains based on gyrB gene sequences. AB - Different strains related to Vibrio splendidus have been associated with infection of aquatic animals. An epidemiological study of V. splendidus strains associated with Crassostrea gigas mortalities demonstrated genetic diversity within this group and suggested its polyphyletic nature. Recently 4 species, V. lentus, V. chagasii, V. pomeroyi and V. kanaloae, phenotypically related to V. splendidus, have been described, although biochemical methods do not clearly discriminate species within this group. Here, we propose a polyphasic approach to investigate their taxonomic relationships. Phylogenetic analysis of V. splendidus related strains was carried out using the nucleotide sequences of 16S ribosomal DNA (16S rDNA) and gyrase B subunit (gyrB) genes. Species delineation based on 16S rDNA-sequencing is limited because of divergence between cistrons, roughly equivalent to divergence between strains. Despite a high level of sequence similarity, strains were separated into 2 clades. In the phylogenetic tree constructed on the basis of gyrB gene sequences, strains were separated into 5 independent clusters containing V. splendidus, V. lentus, V. chagasii-type strains and a putative new genomic species. This phylogenetic grouping was almost congruent with that based on DNA-DNA hybridisation analysis. V. pomeroyi, V. kanaloae and V. tasmaniensis-type strains clustered together in a fifth clade. The gyrB gene-sequencing approach is discussed as an alternative for investigating the taxonomy of Vibrio species. PMID- 15109136 TI - Identification and characterisation of the fur genes in Photobacterium damselae ssp. piscicida and ssp. damselae. AB - The gene encoding the ferric uptake regulator protein (fur gene) of the fish pathogenic bacterium Photobacterium damselae ssp. piscicida Strain D121 was partially amplified using degenerate oligonucleotides. Complete sequencing of the fur gene and neighbouring DNA was accomplished by primer walking. An open reading frame of 447 bp, coding for a protein of 148 amino acids, and with high homology to previously described Fur proteins, was identified. The fur gene of P. damselae ssp. damselae ATCC 35083 was subsequently amplified by PCR with specific primers and its sequence determined, showing a 99.3% similarity to the P. damselae ssp. piscicida fur gene. The P. damselae fur gene was able to complement the fur mutation of Escherichia coli Strain H1681 in an iron-dependent fashion. PMID- 15109137 TI - Comparative study of X-ray computerised tomography and conventional X-ray methods in diagnosis of swimbladder infection in eels caused by Anguillicola crassus. AB - To date, swimbladder lesions due to Anguillicola crassus infection of the European eel Anguilla anguilla have so far been studied only by conventional X ray methods. This is the first study to report the use of computerised tomography (CT) for studying lesions induced by anguillicolosis. Of 50 eels caught by electrofishery from Lake Balaton, Hungary, in autumn 2002 and pre-selected by a conventional X-ray method, 22 specimens were examined with a Siemens Somatom Plus S40 spiral CT scanner. Tomograms, radiographs and photographs of 5 of these, showing anguillicolosis-induced swimbladder lesions of varying severity, are presented. Computerised tomograms provide information on the inner structure, air content and wall thickness of the swimbladder as well as on the number of worms it contains. When the swimbladder is not severely affected or not completely filled with worms, computerised tomography provides adequate data on the shape of the swimbladder, thickness of the swimbladder wall and the location of worms in the lumen. However, in more severe cases, i.e. when the swimbladder is tightly packed with worms or contains no air as a result of wall-thickening, this method fails to determine the number and location of helminths or the thickness of the swimbladder wall. PMID- 15109138 TI - X-cells in fish pseudotumors are parasitic protozoans. AB - Bottom-dwelling teleosts, particularly flatfishes or cod living in temperate to cold seawater, sometimes develop tumor-like lesions on the body surface or in the branchial cavity. These lesions usually contain masses of so called 'X-cells' of unknown origin. We amplified a gene for small subunit ribosomal RNA (18S rRNA) from X-cell lesions of the flathead flounder Hippoglossoides dubius. Phylogenetic analysis clearly classified the obtained sequence as a protozoan, although the organism had no clear affinity with any known protistan groups. In situ hybridization showed that probes specific for the protozoan 18S rRNA hybridized only with X-cells, and not with the host-fish cells, indicating that X-cells harbor the protozoan rRNA. On the other hand, a probe specific for vertebrate 18S rRNA hybridized with the host-fish cells, but not with X-cells. This is conclusive evidence that X-cells are parasitic protozoans. PMID- 15109139 TI - Susceptibility of Baltic and East Atlantic salmon Salmo salar stocks to Gyrodactylus salaris (Monogenea). AB - The susceptibility of a Baltic salmon stock Salmo salar (Indalsalv, central Sweden) to Norwegian Gyrodactylus salaris (Figga strain, central Norway) was experimentally tested and compared with previously obtained results on East Atlantic salmon (Lierelva, SE Norway). Contrary to expectation, the Baltic salmon, which had no prior exposure to this parasite strain, appeared almost as susceptible as the Norwegian salmon parr that naturally experience G. salaris induced mortality. Individually isolated salmon of both stocks sustained G. salaris infections with little evidence of innate resistance. A few individuals of the Indalsalv stock controlled their infection from the beginning, but overall there was considerable heterogeneity in the course of infection in both stocks. On individual hosts, G. salaris growth rates declined steadily throughout the infection, a trend which was particularly marked amongst the Lierelva stock. On shoaling Lierelva fish, there was some evidence of reduced parasite population growth towards the end of the infection; this was not apparent in Indalsalv fishes. These results reflect a growing awareness that not all Baltic salmon may be resistant to Norwegian G. salaris, and that Norwegian and Baltic G. salaris strains may differ in virulence. Consequently, management decisions concerning this parasite-host system should be based upon the actual, and tested, susceptibility of stocks under consideration and not upon identification of stocks as either Atlantic or Baltic. PMID- 15109140 TI - Parasitic infection of sole Solea solea by Prosorhynchus spp. metacercariae (Digenea, Bucephalidae) in Atlantic nurseries under mussel cultivation influence. AB - Cysts of metacercariae were obtained on 2 dates from juvenile sole Solea solea sea-sampled in an area of mussel cultivation (Pertuis Charentais, Bay of Biscay, France). An initial assessment of parasite genus and infestation level was based on 192 cysts extracted from 2 fish samples, taken in August (n = 20) and December 2000 (n = 14). Our results confirmed the sole as second intermediate host of bucephalid trematodes of the genus Prosorhynchus, which has not previously been noticed in Atlantic stocks. Prevalence, ca. 65% on both dates, indicated an substantial infestation of these small fish, with a mean abundance of parasites increasing from August (3.3 +/- 1.1) to December (8.1 +/- 3.4). Cysts were localised in all body parts of the host, and positioning varied depending on sampling date. However, the cephalic area was always the most infested (72.7 and 49.1% in August and December, respectively). Parasite measurements suggested a protracted infestation process, which may be initiated in spring during sole settlement. Most of the largest metacercariae had the rhynchus characteristics of P. crucibulum, though the possibility of them being other species (P. squamatus, P. aculeatus) could not be excluded. As Mytilus edulis is the first intermediate host of Prosorhynchus spp., possible relationships between mussel culture and sole parasitosis are discussed. PMID- 15109141 TI - Impact of a water temperature shift on xenoma clearance and recovery time during a Loma salmonae (Microsporidia) infection in rainbow trout Oncorhynchus mykiss. AB - Previous studies have modelled the relationship between water temperature and the rate of sporulation as defined by xenoma formation during microsporidial gill disease (MGD) in salmon caused by Loma salmonae. Although offering insight into the epidemiology of MGD, a key unexplored area is the role of temperature in the rate of xenoma dissolution including spore release into the environment, and this is crucial to our ability to model horizontal transmission of MGD within confined net-pen populations of farmed salmon. Results from a previous trial suggested that xenoma dissolution may be dramatically hastened as water temperature declines, thus introducing a critical anomaly into any predictive exercise. The data generated herein was evaluated using the statistics of survival analysis to re-establish the baseline relationship of xenoma formation and dissolution relative to water temperature and to compare these results with those of previous studies. We infected 30 individuals of Oncorhynchus mykiss (Walbaum) with macerated xenoma-laden gill material, and afterwards allocated them to tanks with water temperatures of 11, 15, or 19 degrees C and monitored them through a disease cycle. Xenoma onset and clearance times were similar to previous findings with both events being accelerated at higher water temperatures, thereby suggesting a similar temperature response in the current strain to those used in previous studies. Another group of 45 fish was infected with L. salmonae and held at 15 degrees C until xenomas formed, and were subsequently shifted to 11, 15, or 19 degrees C. The median xenoma dissolution time in these tanks was 49, 35 and 28 d, respectively, similar to rates observed when water temperature remained constant. Thus we rejected the hypothesis that a sudden change in water temperature triggers rapid or anomalous xenoma dissolution. PMID- 15109142 TI - Immunohistochemistry, histopathology and ultrastructure of Gasterosteus aculeatus tissues infected with Glugea anomala. AB - Immunohistochemical and histopathological studies were conducted on a population of 3-spined sticklebacks Gasterosteus aculeatus (L.) from Loch Airthrey (Stirling, Scotland) naturally infected with the microsporean Glugea anomala (Moniez 1887). Of the 55 host specimens that were examined, 16 (29.09%) were infected, the intensity of infection ranging from 1 to 4 xenomas per fish, which were principally located within the central portion of the body lateral flank musculature. All 32 G. anomala xenomas examined were mature, their diameter ranging from 936 to 2232 Pum, and their walls of presented a laminar structure. Subcutaneously situated xenomas protruded from the fish body surface, whilst xenomas encountered within the intestine were seen to cause distortion. Light and electron microscopical observations confirmed a host cellular reaction around the xenoma, seen by the presence of eosinophile granule cells (EGCs), and some neutrophils. The occurrences of rodlet cells among the intestinal epithelial cells, and in close proximity to the xenoma wall, were observed in certain specimens. Outside the xenoma wall, macrophage aggregates (MAs) were commonly encountered. Within the xenoma wall, the presence of eosinophile granular cells immunoreactive to the anti-serotonin serum was also recorded. Further immunohistochemical tests revealed that a high number of nerve fibres running along the white lateral muscle fibres were immunoreactive to bombesin-, galanin-, and leu-enkephalin-antisera. Nerve fibres containing bombesin- and leu-enkephalin like substances were also observed in the connective inflammatory tissue around the protozoan cyst, while neurons in the spinal ganglia were immunoreactive to met-enkephalin, and serotonin antisera. The control for the specificity of immunohistochemical reactions was performed using preabsorption tests of each antiserum with the corresponding antigen, and no immunoreactivity was noticed. The data presented are discussed in relation to the occurrence of G. anomala, which alters the pattern of nerve fibres present in the host. Specifically, the protozoan induces a response in the stickleback nervous system, the reaction of which is revealed through the application of immunohistochemical techniques. PMID- 15109143 TI - Growth and development of Argulus coregoni (Crustacea: Branchiura) on salmonid and cyprinid hosts. AB - The obligate fish ectoparasite Argulus coregoni is strictly specific to salmonids and is very rarely found on other fish species. The ability of the parasite to grow and complete its life cycle on a cyprinid host, Rutilus rutilus, was compared with that on a typical salmonid host, Oncorhynchus mykiss. Rearing experiments were run for 42 d with newly hatched metanauplii in flow-through tanks. Body length and sex of the parasites were recorded every 5 d. Growth rates on O. mykiss exceeded those on R. rutilus from the age of 2 wk, at which time the parasites reached a length of about 3.5 mm. Males grew faster than females at the beginning of the experiment up to a length of 2.5 to 3.0 mm; thereafter, a faster growth rate was observed in females. In another experiment, association of parasites with the hosts was monitored and residence time defined as the period between attachment and first detachment from the host. Longer residence time was observed on O. mykiss than on R. rutilus; female parasites stayed on both fish species longer than did males. Faster growth of parasites could be associated with longer uninterrupted periods of attachment to hosts, since frequent detachment means higher energy losses and less time available for feeding. Despite its slower growth on R. rutilus, A. coregoni matured and laid egg clutches, but took 5 d longer than on O. mykiss. The potential of A. coregoni to complete its life cycle on cyprinids could have important ecological consequences, creating an infection reservoir when the main salmonid hosts are rare or temporarily missing. PMID- 15109144 TI - Effects of the parasite Probopyrus ringueleti (Isopoda) on glucose, glycogen and lipid concentration in starved Palaemonetes argentinus (Decapoda). AB - Effects of the branchial ectoparasite isopod Probopyrus ringueleti on the utilization of glucose, glycogen and total lipids on starved shrimp Palaemonetes argentinus were evaluated, as well as the isopod responses to host starvation. Shrimp were maintained for 12 d under laboratory conditions. Parasitized and unparasitized shrimp were starved for 15 d, and glucose, glycogen and total lipid concentrations were determined at 0, 24, 72, 168 and 360 h of starvation. During starvation, control animals (unparasitized) preferentially metabolized lipids and preserved their carbohydrate reserves. Parasitized shrimp had lower lipid concentrations than unparasitized shrimp, and preferentially metabolized carbohydrates. Bopyrids displayed a similar response, with glycogen depleted at the beginning of the starvation period followed by subsequent reestablishment. Our results demonstrated that bopyrids affect host energy allocation. The lower initial lipid concentration of parasitized shrimp suggested that the host is disadvantaged from both food deprivation and isopod nutritional demands. A possible dependency of P. ringueleti on the mechanisms that control host metabolic processes was also suggested. PMID- 15109145 TI - Excretory calcinosis: a new fatal disease of wild American lobsters Homarus americanus. AB - A significant number of moribund and dead lobsters Homarus americanus were reported to New York state authorities by lobster fishers in Long Island Sound (LIS) during the summer of 2002. Morbid lobsters were characterised by an orange discolouration of the abdomen, lethargy, an excess of epibionts and poor post capture survival. On necropsy, severe extensive multifocal or diffuse mineralised granulomatous inflammation of the gills and antennal glands was the most striking pathology. In the gills, granulomas often occluded the lumen of filaments, resulting in congestion, ischemia and coagulative necrosis of gill tissues. In the antennal glands, granulomas were concentrated along the border between the coelomosac and labyrinth. No significant pathogens were recovered from diseased individuals. In prechronic individuals, however, it was evident that granulomas were focused around calcium carbonate (aragonite) crystals. This disease may result from anomalously high sea-bottom temperatures in LIS (approximately 23 degrees C) during the summer of 2002 and associated disruptions of the calcium chemistry of lobsters in favour of deposition of minerals in soft tissues. The ultimate cause of death of affected lobsters is probably respiratory failure due to reduced effective surface area of the gills, exacerbated by hypermetabolic temperatures and an abundance of epibionts. PMID- 15109146 TI - Pathogenicity testing of shellfish hatchery bacterial isolates on Pacific oyster Crassostrea gigas larvae. AB - Bacterial diseases are a major cause of larval mortality in shellfish hatcheries. Even with proper sanitation measures, bacterial pathogens cannot be eliminated in all cases. The pathogenicity of bacteria isolated from Pacific Northwest shellfish hatcheries to Pacific oyster Crassostrea gigas larvae was investigated. We found 3 highly pathogenic strains and 1 mildly pathogenic strain among 33 isolates tested. These strains appear to be members of the genus Vibrio. Although there have been many studies of bivalve bacterial pathogens, a standard method to assess bacterial pathogenicity in bivalve larvae is needed. Thus, we developed 2 methods using either 15 ml conical tubes or tissue culture plates that were employed for rapidly screening bacterial strains for pathogenicity to Pacific oyster larvae. The tissue culture plates worked well for screening both mildly pathogenic strains and LD50 (lethal dose) assays. This method allowed for non intrusive and non-destructive observation of the oyster larvae with a dissecting microscope. The LD50 for the 3 highly pathogenic strains ranged between 1.6 and 3.6 x 10(4) colony forming units (CFU) ml(-1) after 24 h and between 3.2 x 102 and 1.9 x 10(3) CFU ml(-1) after 48 h. PMID- 15109147 TI - Perkinsus mediterraneus n. sp., a protistan parasite of the European flat oyster Ostrea edulis from the Balearic Islands, Mediterranean Sea. AB - A new species, Perkinsus mediterraneus, a protistan parasite of the European oyster Ostrea edulis (L.), farmed along the coast of the Balearic Islands, Mediterranean Sea, is described. Morphological examinations with light and transmission electron microscopy, DNA sequence-analysis and enlargement in Ray's fluid thioglycollate medium (RFTM) confirmed that this parasite belongs to the genus Perkinsus. Specific morphological and genetic characteristics indicated that it should be considered a new species in the genus. Sequencing of the small subunit ribosomal (ssu rRNA) gene confirmed that the parasite belongs to the genus Perkinsus, and sequences of the internal transcribed spacer (ITS) were distinct from any Perkinsus ITS sequences previously published and/or deposited in the GenBank. Phylogenetic analysis revealed that the ITS sequences of the new species formed a monophyletic group comprising a sister clade to the P. atlanticus/olseni group. In addition, morphological differences were observed between the new species and the other described Perkinsus spp.. After incubation in RFTM for 1 wk, the prezoosporangium had reached an extremely large size (97.4 +/- 1.99 microm) (mean +/- SE), and after 2 wk incubation had again almost doubled in size (167.1 +/- 8.09 microm). The discharge-tube length was one sixth the diameter of the zoosporangium, i.e. a ratio of 17.36:97.38, the lowest ratio observed for any Perkinsus species. At the ultrastructural level, zoosporangia and zoospores exhibited some differences compared to other Perkinsus species. PMID- 15109148 TI - Metastatic squamous cell carcinoma in two loggerhead sea turtles Caretta caretta. AB - We received 2 stranded loggerhead sea turtles (Caretta caretta) with squamous cell carcinomas to necropsy. The dead turtles had been collected in Gran Canaria and Fuerteventura in April 1994 and May 1997, respectively to determine the cause of death. One turtle had 3 ulcerated lesions in the dorsal part of the neck and several irregular masses in the lungs and kidneys. Histologic examination of lesions in the skin, lungs, kidneys, and ventricular myocardium revealed neoplastic proliferation of abnormal keratinocytes. Ultrastructural examination identified the tumoral cells as epithelial cells. The second turtle had 4 lesions in the skin of the head and flippers, and several irregular masses in the lungs, liver, and kidneys. Histological examination revealed a squamous cell carcinoma with metastases to muscle tissue, liver, lungs, and kidneys. Attempts to characterize the tumoral cells by immunohistochemistry using several monoclonal and polyclonal antisera against high and low molecular weight cytokeratins from mammals, as well as vimentin and desmin, failed. Differences between reptilian keratins (mainly beta-keratins) and mammalian keratins (mainly alpha-keratins) could explain this absence of immunoreactivity. This is the first description of squamous cell carcinoma in sea turtles. PMID- 15109149 TI - Solar UV radiation does not inactivate marine birnavirus in coastal seawater. AB - We examined the inactivation kinetics of marine birnavirus (MABV) in a coastal sea, in seawater samples collected from 50 cm depth. MABV was added to both natural and autoclaved seawater at a concentration of 6 x 10(6.43) TCID50 (50% tissue culture infectious dose) ml(-1), put in dialysis tubes and incubated at the original depth. The inactivation of MABV by solar UV radiation was examined using light and dark tubes. The infectivity titer of MABV was measured by the TCID50 method using CHSE-214 cells. Virus infectivity in natural seawater decreased quickly and was below the detection limit by 270 min in both light and dark conditions; however, virus infectivity was maintained in the autoclaved seawater until 420 min. These results suggest that the loss of virus infectivity is not caused by sunlight UV radiation. PMID- 15109150 TI - Comparison of toxic effects of nitric oxide and peroxynitrite on Uronema marinum (Ciliata: Scuticociliatida). AB - To discover the effects of nitric oxide (NO) and peroxynitrite on Uronema marinum (a ciliate responsible for systemic scuticociliatosis in cultured olive flounder Paralichthys olivaceus), the dose-dependent inhibitory effect of NO donors, S nitroso-N-acetylpenicillamine (SNAP) and 3-morpholinosydnonimine (SIN-1) on the proliferation and survival of U. marinum was investigated. The inhibitory effects of exogenous superoxide dismutase (SOD) and catalase on the toxicity of SIN-1 were also investigated. After 24 h of incubation in the presence of 0.2 mM SNAP, the number of ciliates was not statistically different from that of the controls, whereas incubation in the presence of 0.5 mM SNAP reduced the number of parasites significantly to 59.1% of controls. Concentrations of SNAP higher than 0.5 mM resulted in greater reductions in the number of ciliates, but levels of generated NO far exceeded physiological ranges. The number of viable ciliates incubated for 24 h with 0.2 mM SIN-1 was reduced significantly to 25.0%, and all ciliates were killed by incubation in concentrations above 0.5 mM SIN-1. Although SOD decreased the toxic effect of SIN-1 on U. marinum, protection was not complete and did not improve after increasing the SOD concentration from 50 to 400 U ml(-1). Addition of catalase ranging from 500 to 10000 U ml(-1) completely protected U. marinum from SIN-1 toxicity. Ciliates exposed to catalase alone or catalase plus SIN-1 showed significantly higher and dose-dependent proliferation rates compared to controls. Addition of haemoglobin, ranging from 0.5 to 2.0 mg ml(-1), also protected U. marinum from SIN-1 toxicity, and increased the proliferation rate dose-dependently. In conclusion, resistance of U. marinum to oxidative and nitrative stress may allow this pathogen to withstand the NO- and oxygen-radical dependent killing mechanisms of phagocytic cells. PMID- 15109151 TI - First record of the microsporidian parasite Steinhausia mytilovum in Mytilus sp. (Bivalvia: Mytilidae) from France. AB - Steinhausia mytilovum is a globally distributed microsporidian parasite which infects the oocytes of the blue mussels Mytilus edulis and M. galloprovincialis. Despite the intensive monitoring effort made on mussel populations, the parasite has not previously been reported in France. We report herein on the occurrence of S. mytilovum in Mytilus sp. from 1 cultured and 2 natural populations on the northern coast of France, thus extending the parasite's known distribution northwards. We also report on the observation in 1989 of S. mytilovum in M. galloprovincialis from the Golfe de Fos area in the Mediterranean Sea (South of France). S. mytilovum was observed in the European hybrid zone between M. edulis and M. galloprovincialis, which therefore renders the exact taxonomic status of the infected hosts unknown. The prevalence of the parasite was low, which suggests that its effect on mussel populations was probably limited. PMID- 15109152 TI - Sinergasilus polycolpus, a new copepod species in the ichthyoparasitofauna of Serbia and Montenegro. AB - The parasitic copepod Sinergasilus polycolpus was identified on the gills of bighead carp Aristichthys nobilis from 2 localities (Kladovo and Slankamen) in the Serbian part of the River Danube. This parasite is species-specific for 2 Chinese carp, the bighead carp and the silver carp Hypophthalmichthys molitrix. It was accidentally introduced into Serbia and Montenegro together with fry of these herbivorous carp intended for aquaculture and control of phytoplankton blooms. There is no record in the available literature of this parasite for European freshwaters. Our identification of S. polycolpus signals the possible spread of the infectious disease sinergasilosis in natural freshwaters and in fishponds, similar to bothriocephalosis, caused by Bothriocephalus opsariichthydis, which was introduced with the fry of various herbivorous species from the Amour River basin (USSR) into almost all countries throughout the world. PMID- 15109153 TI - The founding of the Edinburgh medical school. AB - This article seeks to show that the usual accounts of the founding of the Edinburgh Medical Faculty in 1726 give undue prominence to John Monro, an Edinburgh surgeon, and to George Drummond, later Lord Provost of Edinburgh. They do so because their authors have ignored the ways in which patronage appointments, such as medical professorships, were and had been dispensed in the city of Edinburgh and in its university. There the Town Council was only nominally independent when it came to making professors. Medical historians have been equally cavalier in their treatment of the roles of leading politicians, especially of Archibald Campbell, first Earl of Ilay and later third Duke of Argyll, who was the most important Scottish politician working between c. 1716 and his death inl 1761. A more realistic view of the history of Scottish medicine would not ignore the realities of politics and the relation of these to institutions, such as the Edinburgh Medical Faculty. PMID- 15109154 TI - Anatomists and entrepreneurs in early eighteenth-century London. AB - Anatomical demonstration in the eighteenth century took place in many formats. In this essay I discuss public anatomical demonstration as performed by entrepreneurial anatomists in London between 1700 and 1740. These anatomists offered courses, advertised in newspapers, to anyone who was willing to pay. In contrast to courses offered in official settings to prospective physicians and surgeons, these courses emphasized natural philosophy and natural theology rather than practical knowledge. Entrepreneurial lecturers also aimed to entertain. In this article I examine the lectures of James Douglas, William Cheselden, and Frank Nicholls, each of whom differed significantly from the others in style and content but were all anatomical entrepreneurs. All of them, moreover, employed not only human cadavers but also living and dead animals in their lessons. I examine the content of the lectures and the motivations of the lecturers' audiences. I also argue that the prevailing historiographical representation of eighteenth-century science as "polite" requires considerable revision to accommodate as impolite an activity as public anatomy. PMID- 15109155 TI - A case of Parturiunt montes, nascetur ridiculus mus? the BMA Nutrition Committee 1947-1950 and the political disengagement of nutrition science. AB - During spring 1947, there was a controversy about the consequences for health of food rationing in Britain; among the issues at stake being the accusation that government departments were failing to release relevant data. The British Medical Association (BMA) responded by establishing a Nutrition Committee to investigate the effects of rationing. Leading members of the BMA expected the committee to intervene decisively in the debate, presenting the medical profession as protectors of the people. The committee, however, was dominated by officials and others who were concerned with presenting the government's record in a good light. Furthermore, by the time the committee reported, the food situation had improved. In the event, the BMA's Nutrition Report was deemed a success if it was received quietly by the press. The deliberations of the committee had involved the controlled release of government data to interested parties, and took place during a period of transition in the government's work in nutrition, from comprehensive emergency intervention toward routine peacetime surveillance. By providing a new set of standards that were used for measuring the adequacy of diets as revealed by the annual National Food Survey, the report contributed to the establishment of the postwar modus operandi of nutrition science and the peacetime system for monitoring the health of the nation. PMID- 15109156 TI - Good figures. PMID- 15109157 TI - Analyses of facial attractiveness on feminised and juvenilised faces. AB - We conducted two experiments to investigate the psychological factors affecting the attractiveness of composite faces. Feminised or juvenilised Japanese faces were created by morphing between average male and female adult faces or between average male (female) adult and boy (girl) faces. In experiment 1, we asked the participants to rank the attractiveness of these faces. The results showed moderately juvenilised faces to be highly attractive. In experiment 2, we analysed the impressions the participants had of the composite faces by the semantic-differential method and determined the factors that largely affected attractiveness. On the basis of the factor scores, we plotted the faces in factor spaces and analysed the locations of attractive faces. We found that most of the attractive juvenilised faces involved impressions corresponding to an augmentation of femininity, characterised by the factors of 'elegance', 'mildness', and 'youthfulness', which the attractive faces potentially had. PMID- 15109158 TI - Familiarity breeds attraction: effects of exposure on the attractiveness of typical and distinctive faces. AB - Several studies have shown that facial attractiveness is positively correlated with both familiarity and typicality. Here we manipulated the familiarity of typical and distinctive faces to measure the effect on attractiveness. In our first experiment, we collected ratings of attractiveness, distinctiveness, and familiarity using three different groups of participants. Our stimuli included 84 images of female faces, presented in a full-face view. We replicated the finding that attractiveness ratings negatively correlate with distinctiveness ratings. In addition, we showed that attractiveness ratings were positively correlated with familiarity ratings. In our second experiment, we demonstrated that increasing exposure to faces increases their attractiveness, although there was no differential effect of exposure on typical and distinctive faces. Our results suggest that episodic familiarity affects attractiveness ratings independently of general or structural familiarity. The implications of our findings for the 'face space' model are discussed. PMID- 15109159 TI - Demonstrating the acquired familiarity of faces by using a gender-decision task. AB - We report an experiment in which participants made gender judgments (male or female) to faces. There were three groups of faces: unfamiliar, familiar (celebrities), and a set which had been learned earlier by the participants during the experimental session. The principal purpose of this study was to establish an indirect measure of assessing whether faces have become familiar through learning that does not require overt recognition. Bruce and Young's (1986 British Journal of Psychology 77 305-327) model of face recognition suggests face processing tasks are independent of one another and so familiarity should have no impact on the time taken to perform gender decisions. However, recent studies have suggested that some face processes are not completely independent. A gender judgment is a simple task which would be useful in face-learning experiments. We examined whether exposure to previously novel faces facilitates a later gender decision to those faces. During a learning stage, participants viewed a set of unfamiliar faces. At test, participants were able to assign gender faster to previously familiar (famous) faces and learned faces than they were to unfamiliar faces. Therefore familiarity can influence the speed at which gender is analysed. We explain our findings with reference to the Burton et al (1990 British Journal of Psychology 81 361-380) interactive activation and competition (IAC) model of face recognition and discuss how the gender judgment might be employed as a means of tracking the acquisition of familiarity in face-learning studies. PMID- 15109160 TI - Ethnic categorisation of faces is not independent of face identity. AB - Is the extraction of a visually derived semantic code from faces (ethnicity) affected by face identity (familiarity) or not? The traditional view considers that this operation is made independently of face identity, and in parallel with the recognition of identity. However, some recent studies cast doubt on this parallel thesis regarding other visually derived semantic codes, namely: facial expression, facial speech, apparent age, and gender. Twenty-eight Caucasian participants were enrolled in an 'ethnic-decision' task on morphed faces made of an Asiatic source face and a Caucasian source face, in the proportion of 70%-30%. Half of the original faces were previously made familiar by a learning procedure (associating the face, surname, occupation, and city of residence of the person displayed), while the remaining half were unfamiliar. The results showed clearly that ethnic decision was affected by face familiarity. This adds support to the thesis according to which the identification of identity and the extraction of visually derived semantic codes are not made independently from each other and that the 'parallel-route' hypothesis becomes weakly supported. PMID- 15109161 TI - Configuration-specific attentional modulation of flanker- -target lateral interactions. AB - Elements of a contour are often easier to detect when they possess collinearity, with their local orientations matching the global orientation of the contour. We recently reported attentional modulation of such lateral interactions between a central near-threshold target Gabor patch and flanking high-contrast patches (Freeman et al, 2001 Nature Neuroscience 4 1032-1036). Here, we examined whether such attentional effects reflect specific modulation of mechanisms sensitive to collinear configurations, or instead more general modulation of sensitivity to either the global or local orientation-components of the stimulus. Thresholds for detecting a central Gabor target were measured, while observers also judged the Vernier alignment between one pair of flankers and ignored a second flanker pair (when present). Target contrast-thresholds were facilitated only when attending collinear flankers. There was no facilitation when attending flankers that shared only local orientation with the target, or flankers that fell on a global axis aligned with target orientation but having orthogonal local orientation. Ignored collinear flankers had no effect on target thresholds. These results demonstrate strong and specific attentional modulation of contour-integration mechanisms in early vision sensitive to collinear configurations. PMID- 15109162 TI - Does previewing one stimulus feature help conjunction search? AB - We examined the effects of previewing one aspect of a search display, in order to determine what subset of display information is most useful as a prelude to a search task. Observers were asked to indicate the presence or absence of a known target, in a conjunction search where the target was defined by the combination of colour and orientation (a yellow horizontal line presented among yellow vertical and pink horizontal distractors). In the colour preview condition of experiment 1, observers were first shown a 1 s preview of the locations and colours of the search items before the actual search set was presented. That is, search items first appeared as yellow and pink squares for 1 s, which each then turned into yellow and pink oriented lines (in the same locations) which comprised the display to be searched. In the orientation preview condition, observers were first shown a 1 s preview of the locations and orientations of the search items before the actual search display was presented. These two conditions were compared to a control condition consisting of standard conjunction search without any preview display. There was no effect of colour preview; there was a marginal effect of orientation preview, but in the opposite direction from what was expected reaction time increased for orientation preview searches. In experiment 2 these previews were compared to two spatial cueing conditions; in this experiment the colour preview did provide a small amount of help. Finally, in experiment 3 both previews were presented in succession, and increased facilitation was found, in particular when the colour preview preceded the orientation preview. These findings are discussed in relation to the literature, in particular the Guided Search model (Wolfe et al 1989 Journal of Experimental Psychology: Human Perception and Performance 15 419-433; Wolfe 1994 Psychonomic Bulletin & Review 1 202-238). PMID- 15109163 TI - Revisiting Snodgrass and Vanderwart's object pictorial set: the role of surface detail in basic-level object recognition. AB - Theories of object recognition differ to the extent that they consider object representations as being mediated only by the shape of the object, or shape and surface details, if surface details are part of the representation. In particular, it has been suggested that color information may be helpful at recognizing objects only in very special cases, but not during basic-level object recognition in good viewing conditions. In this study, we collected normative data (naming agreement, familiarity, complexity, and imagery judgments) for Snodgrass and Vanderwart's object database of 260 black-and-white line drawings, and then compared the data to exactly the same shapes but with added gray-level texture and surface details (set 2), and color (set 3). Naming latencies were also recorded. Whereas the addition of texture and shading without color only slightly improved naming agreement scores for the objects, the addition of color information unambiguously improved naming accuracy and speeded correct response times. As shown in previous studies, the advantage provided by color was larger for objects with a diagnostic color, and structurally similar shapes, such as fruits and vegetables, but was also observed for man-made objects with and without a single diagnostic color. These observations show that basic-level 'everyday' object recognition in normal conditions is facilitated by the presence of color information, and support a 'shape + surface' model of object recognition, for which color is an integral part of the object representation. In addition, the new stimuli (sets 2 and 3) and the corresponding normative data provide valuable materials for a wide range of experimental and clinical studies of object recognition. PMID- 15109164 TI - The influence of object size and surface shape on shape constancy from stereo. AB - The failure of shape constancy from stereoscopic information is widely reported in the literature. In this study we investigate how shape constancy is influenced by the size of the object and by the shape of the object's surface. Participants performed a shape-judgment task on objects of five sizes with three different surface shapes. The shapes used were: a frontoparallel rectangle, a triangular ridge surface, and a cylindrical surface, all of which contained the same maximum depth information, but different variations in depth across the surface. The results showed that, generally, small objects appear stretched and large objects appear squashed along the depth dimension. We also found a larger variance in shape judgments for rectangular stimuli than for cylindrical and ridge-shaped stimuli, suggesting that, when performing shape judgments with cylindrical and ridge-shaped stimuli, observers rely on a higher-order shape representation. PMID- 15109165 TI - Orientation-selective adaptation during motion-induced blindness. AB - When a global moving pattern is superimposed on high-contrast stationary or slowly moving stimuli, the latter occasionally disappear for periods of several seconds (motion-induced blindness, MIB). Here, an adaptation paradigm was used to determine if orientation-selective adaptation still occurs for the stimulus that is no longer visible. Two slowly drifting high-contrast Gabor patches were presented to observers. As soon as both patches disappeared, one was eliminated from the screen. After 2 s, two low-contrast Gabor patches were presented as tests at the same locations and observers were asked to report their orientations. The observers' performance was significantly higher when the orientation of the low-contrast test patch was orthogonal to the orientation of the high-contrast adapting patch (p < 0.0001) for the location where the patch was present during MIB, even though it was perceptually invisible. The observers' performance was not significantly different at the adjacent control location where the stimulus was absent during the MIB. Although no stimulus was visible at either location, orientation-selective adaptation was preserved only for the location at which the patch remained present. Since orientation information is processed in low-level visual areas such as the primary visual cortex (V1), we conclude that MIB originates in an area higher than V1. PMID- 15109166 TI - [Syphilis]. PMID- 15109167 TI - [How to help children and adolescents suffering from cancer]. PMID- 15109168 TI - [Chronic lymphocytic leukemia]. AB - The last years have yielded important new insights into the biological characteristics of chronic lymphocyte leukemia cells, which have lend to modify the diagnosis and prognosis criteria of the disease. The diagnosis is now based on morphology and immunophenotyping of blood lymphocytes, making the bone marrow examination unnecessary. However, detection of a clonal expansion of B lymphocytes in the blood is far to be synonymous of leukemia, and even less an argument for starting a treatment. Prognosis classifications based on clinical and blood counts are now refined with genotyping and cytogenetics. These new tools should improved the therapeutic strategies in the next future. PMID- 15109169 TI - [Syphilis: the stakes of a reappearance]. PMID- 15109170 TI - [Syphilis, new epidemiologic features]. AB - The epidemiologic survey by the InVS from data from voluntary sites shows an increase of the cases of syphilis from 2000 to 2002 in France. Most of patients with early syphilis are men having sex with men and more than half of cases is HIV infected patients. Resurgence of syphilis reflects the relapse of high-risk sexual practices and the most sexual practice evocated is unprotected oral sex. The same situation has been reported in countries from the west of Europe. In east of Europe, the epidemiology is a little bit different with an increase of cases since the early 90's mostly in sex workers and in relation to the rapid increase in the number of injecting drug users. In United States, although increases in syphilis rates among men who have sex with men have been documented since 1997, the majority of cases have been reported in the South affecting disproportionately blacks, reflecting low access to medical care but also racial segregation in sexual activity. The resurgence of syphilis while it is limited highlights the need for more health promotion initiatives especially in the most affected population. PMID- 15109171 TI - [Syphilis: clinical, biological and therapeutical aspects]. AB - Syphilis is a STI. The present epidemic is related to the relapse into unsafe sexual behaviour. HIV has not modified clinical aspects and the great simulator implies to perform TPHA and VDRL in various circumstances. Treatment is simple and consensual in early syphilis. Treatment of late syphilis is dependent upon indications of CSF control, to be discussed by specialists, particularly in HIV+ patients. PMID- 15109172 TI - [Atlas: clinical traps of syphilis]. PMID- 15109173 TI - [Interpretation of questionable syphilis serologies]. AB - Serologic tests are essential for the diagnosis of syphilis. In symptomatic patients, serodiagnosis confirms the clinical diagnosis. During the asymptomatic stage, interpretation of serologic tests is more difficult, either because of low serologic titres or because of discrepant serologies. Questioning of the patient will help to find a history of syphilis within the previous months or years, an adequate therapy or no treatment at all. Another issue that must be mentioned is the impossibility to differentiate syphilis and non-sexually acquired treponemal infections. PMID- 15109174 TI - [Syphilis and pregnancy]. AB - Consequences of syphilis for mother, pregnancy, fetus and child are considerable, but preventable. Serological screening must be offered at the first prenatal visit, using both a treponemal (eg. TPHA) and a non treponemal (eg. VDRL or RPR) test. When the results are compatible with any type of active syphilis, treatment is required. The treatment of choice is penicillin: benzathine penicillin G, 2.4 million units intramuscular, repeated one week later, and most authors recommend a third dose if a late latent syphilis is suspected, or in case of coinfection with HIV. Women with a proven penicillin allergy can be desensitized. Alternative therapies, such as macrolids, are less well evaluated. Follow-up during and after therapy must not be neglected. In case of Jarisch-Herxheimer reaction, the mother should be managed on an inpatient basis, and the fetus carefully monitored. The VDRL should be repeated (usually every trimester), and therapy be renewed if there is not a significant decrease in titer. The fetus should be followed by serial ultrasound examinations. Finally, the child must be followed up clinically and biologically, and treated in case of congenital syphilis. The residual risk of adverse outcome is increased in case of reinfection, lack of maternal therapy or incomplete treatment, or when diagnosis and therapy are performed late in pregnancy. PMID- 15109176 TI - [Syphilis alert]. PMID- 15109175 TI - [When to think about neurosyphilis?]. AB - The clinical spectrum of neurosyphilis is large. Serologic tests for syphilis are commonly indicated in diseases involving the central nervous system. The diagnosis of syphilis relies on epidemiological, clinical and biological grounds (cerebrospinal fluid abnormalities and RPR test). The treatment of neurosyphilis is still penicillin G. PMID- 15109177 TI - [Assessment: vademecum and implementation]. PMID- 15109178 TI - [Glomerular nephropathies]. PMID- 15109179 TI - [Child abuse and children in danger. Maternal and infant protection]. PMID- 15109180 TI - [Aortic regurgitation]. PMID- 15109181 TI - [Acute diarrhoea in the child and in the adult (with treatment)]. PMID- 15109182 TI - [Children with special needs: the decision on the appropriate type of care and its implementation]. PMID- 15109183 TI - [Diuretic therapy]. PMID- 15109184 TI - [Acute fever in children. Criteria to identify serious illness in febrile children]. PMID- 15109185 TI - [Movies against syphilis: attractive beginnings]. PMID- 15109186 TI - Vital capacity manoeuvre in general anaesthesia: useful or useless? AB - As atelectasis occurs in most patients during general anaesthesia and may be one of the major causes for the development of hypoxaemia and nosocomial pneumonia, its prevention may be considered as an important objective in perioperative management. The major causative mechanisms are the loss of respiratory muscle tone, compression and gas absorption. Vital capacity manoeuvres have been proposed as a means to eliminate atelectasis in the vast majority of patients and restore normal pulmonary gas exchange during general anaesthesia. In this review we describe the pathogenesis of atelectasis in the perioperative period and discuss in the light of recent published investigations the suitability of the vital capacity manoeuvre as a tool during general anaesthesia. Reviewing the current literature, a vital capacity manoeuvre during general anaesthesia may only be useful under specific circumstances when mechanical ventilation with a high inspiratory fraction of oxygen is required or during cardiac surgery at the end of cardiopulmonary bypass to reduce the amount of atelectasis and to maintain adequate gas exchange. PMID- 15109187 TI - A comparison of the effects of ranitidine and omeprazole on volume and pH of gastric contents in elective surgical patients. AB - BACKGROUND AND OBJECTIVE: In cases of aspiration of gastric contents the risk of pneumonitis is dependent on the pH and volume of the gastric contents. Omeprazole and rantidine each decrease gastric volume and increase gastric pH. We evaluated the efficacy of preoperative administration of omeprazole (60 mg) or ranitidine (150 mg) in the prophylaxis of aspiration pneumonitis. METHODS: Data were obtained from 75 elective female surgical patients randomly allocated to one of three groups, who received either omeprazole 60 mg orally, or ranitidine 150 mg orally, or neither, on the evening prior to, and on the morning of, surgery. Gastric volume and pH was measured using blind aspiration. RESULTS: Both pH < 2.5 and volume > 25 mL were present in none of the patients in either the ranitidine or omeprazole groups, compared to 15 of 25 control patients (P < 0.0001). CONCLUSIONS: Preoperative oral administration of omeprazole (60 mg) or ranitidine (150 mg) reduced residual gastric content volume and increased pH > 2.5, possibly reducing the effects of pulmonary aspiration of gastric contents. PMID- 15109188 TI - Conventional stepwise vs. vital capacity rapid inhalation induction at two concentrations of sevoflurane. AB - BACKGROUND AND OBJECTIVE: A multicentre study was conducted to compare three methods of inhalation induction with sevoflurane in adult premedicated patients. METHODS: One-hundred-and-twenty-five adult patients of ASA I-II were scheduled for short elective surgical procedures (< 90 min) under general anaesthesia with spontaneous ventilation of the lungs via a laryngeal mask airway. Patients were randomly assigned to one of three groups: conventional stepwise inhalation induction group (Group C) or vital capacity rapid inhalation induction groups at 4.5% (Group VC4.5) or at 8% sevoflurane (Group VC8). Before anaesthetic induction, fentanyl 1 micro kg(-1) was given and the face mask applied with the anaesthetic breathing system primed with sevoflurane 4.5% or 8% in the respective vital capacity groups. Loss of eyelash reflex, time to cessation of finger tapping, laryngeal mask insertion, side-effects and adequacy of induction were recorded. RESULTS: The time to loss of eyelash reflex was significantly shorter in both vital capacity groups vs. the control group: VC8: 68 +/- 7 s; and VC4.5: 94 +/- 6.5 s vs. C: 118 +/- 6.4s (P < 0.0001). Significant differences were found in all pairwise comparisons for time to cessation of tapping: Group VC8 (62 +/- 7 s), Group VC4.5 (85 +/- 6 s) and Group C (116 +/- 6 s; P < 0.0001). The time to laryngeal mask insertion was significantly shorter in the Group VC8 (176 +/- 13 s) compared with the other two groups, Group VC4.5 (219 +/- 13 s) and Group C (216 +/- 9 s). There were no significant differences in the incidence of side effects between the three groups. CONCLUSIONS: Inhalation induction of anaesthesia with sevoflurane with the three techniques tested is safe, reliable and well accepted by the patients. The vital capacity rapid inhalation group primed with sevoflurane 8% was the fastest method with no relevant side-effects. PMID- 15109189 TI - Postoperative changes in the full-field electroretinogram following sevoflurane anaesthesia. AB - BACKGROUND AND OBJECTIVE: We tested the hypothesis that disturbances of the visual pathway persist following general anaesthesia, even after normal clinical discharge criteria have been met. METHODS: We performed full-field flash electroretinography in the right eye of 10 unpremedicated ASA I patients who underwent N2O/sevoflurane anaesthesia. Electroretinograms were recorded preoperatively, immediately after discharge from the recovery room and 2 h after discontinuation of sevoflurane. The time at which postanaesthesia discharge score first exceeded 9 was also noted. Data were analysed using paired, one-tailed Student's t-test. RESULTS: Latency of the b-wave on the photopic electroretinogram was greater at each postoperative time point (30.5 +/- 0.9 and 30 +/- 1.3 ms), compared to preoperative values (29.2 +/- 0.8 ms, P < 0.001 and P = 0.04, respectively). The A-B amplitude of the b-wave was less postoperatively (220.3 +/- 52.7 and 210.3 +/- 42.7 pV) compared to values before operation (248.1 +/- 57.6 microV, P = 0.03 and P = 0.01, respectively). Oscillatory potential latencies were greater at each postoperative time point (21.4 +/- 0.5 and 20.8 +/ 0.6 ms) compared to before operation (20.4 +/- 0.4 ms, P < 0.001 and P = 0.03, respectively). Oscillatory potential amplitudes were less at the first postoperative time point (17.5 +/- 6.1 microV), compared to preoperative values (22 +/- 6.4 microV, P = 0.04). CONCLUSIONS: Postoperative electroretinogram abnormalities are consistently present in patients who have undergone N2O/sevoflurane anaesthesia. These abnormalities persist beyond the time at which standard clinical discharge criteria have been met. PMID- 15109190 TI - Respiratory effects of the kneeling prone position for low back surgery. AB - BACKGROUND AND OBJECTIVE: The kneeling prone position is often used for low back surgery in order to decrease intraoperative bleeding and increase the surgical exposure of the vertebral canal. The aim of this study was to assess effects of the kneeling prone position on respiratory gas exchange focusing on oxygen consumption and early changes in oxygenation. METHODS: Thirty ASA I-II patients scheduled for low back surgery in the kneeling prone position were studied. Anaesthesia was maintained with isoflurane, 1.2% end-tidal concentration. Respiratory gas exchange was measured with indirect calorimetry. RESULTS: When the patients were turned into the kneeling prone position their oxygenation was immediately improved--measured by arterial oxygen tension and arterial oxygen saturation. The oxygen uptake rate did not change from a baseline supine level of 76 mL min(-1) m(-2), but the carbon dioxide excretion rate decreased from a baseline supine value of 71 mL min(-1) m(-2) to 66 mL min(-1) m(-2) at 5 and 10 min after the kneeling prone position was adopted. Alveolar ventilation decreased in the kneeling prone position. CONCLUSIONS: The present study demonstrates that the kneeling prone position improves oxygenation and that the mechanisms involved are fast in onset. Furthermore, the prone position does not change oxygen consumption although alveolar ventilation is significantly reduced. The changes in alveolar ventilation could possibly be the result of circulatory changes caused by the prone position, but further studies are needed to clarify that hypothesis. PMID- 15109191 TI - Comparison of patient-controlled and operator-controlled conscious sedation for restorative dentistry. AB - BACKGROUND AND OBJECTIVE: The use of midazolam for conscious sedation is an accepted method of anxiety control in restorative dentistry. A lack of predictability in its effects requires the dose of midazolam to be adjusted to individual patient's requirements. We determined whether patient-controlled sedation was a suitable alternative to operator-controlled sedation in restorative dentistry. METHODS: A randomized crossover clinical trial involving 35 consecutive patients undergoing similar dental procedures. Patients were randomly given midazolam, administered either by the patient or by the operator at the first visit and the alternative option on the second visit. All patients were ASA I-II and their ages ranged between 20 and 48 yr. Blood pressure, heart rate, oxygen saturation and patient satisfaction were recorded. RESULTS: The onset time and initial dose for sedation were similar with the two methods of administration and the sedation scores and vital signs were satisfactory. In the patient-controlled group the mean total dose of midazolam was 7.9 (+/- 4.2 SD) mg cf. 4.2 (+/- 1.8 SD) mg in the operator-controlled group (P < 0.05). The time to fitness for discharge (15.4 (+/- 11.9 SD) min) was greater in the patient controlled group cf. the operator-controlled group (8.5 (+/- 9.5 SD) min), P < 0.05. CONCLUSION: This study shows that patient-controlled sedation is a suitable alternative to operator-controlled sedation in the management of anxious dental patients. PMID- 15109192 TI - Changes of jugular venous blood temperature associated with measurements of cerebral blood flow using the transcerebral double-indicator dilution technique. AB - BACKGROUND AND OBJECTIVE: The transcerebral double-indicator dilution technique is a recently developed method to measure global cerebral blood flow at bedside. It is based on bolus injection of ice-cold indocyanine green dye and simultaneous recording of resulting thermo- and dye-dilution curves in the aorta and the jugular bulb. However, with this method 40 mL of ice-cold solution is administered as a bolus. Therefore, this prospective clinical study was performed to elucidate the effects of repeated administration of indicator on absolute blood temperature and on cerebral blood flow and metabolism. METHODS: The investigation was performed in nine male patients scheduled for elective coronary artery bypass grafting. Absolute blood temperature was measured in the jugular bulb and in the aorta before and after repeated measurements using the transcerebral double-indicator dilution technique. RESULTS: During the investigated time course, the blood temperature in the jugular bulb, compared to the aorta, was significantly higher with a mean difference of 0.21 degrees C. The administration of an ice-cold bolus reduced the mean blood temperature by 0.06 degrees C in the jugular bulb as well as in the aorta. After the transcerebral double-indicator dilution measurements a temperature recovery to baseline conditions was not observed during the investigated time period. Cerebral blood flow and cerebral metabolism did not change during the investigated time period. CONCLUSIONS: Repeated measurements with the transcerebral double-indicator dilution technique do not affect absolute jugular bulb blood temperatures negatively. Global cerebral blood flow and metabolism measurements remain unaltered. However, accuracy and resolution of this technique is not high enough to detect the effect of minor changes of physiological variables. PMID- 15109193 TI - Hypoxaemia after cardiac surgery: clinical application of a model of pulmonary gas exchange. AB - BACKGROUND AND OBJECTIVE: To investigate the clinical application of a mathematical model of pulmonary gas exchange, which ascribes hypoxaemia to shunt and ventilation/perfusion mismatch. Ventilation/perfusion mismatch is quantified by deltaPO2, which is the drop in oxygen pressure from alveoli to lung capillaries. Shunt and deltaPO2 were used to describe changes in oxygenation after coronary artery bypass grafting. METHODS: Fourteen patients were studied 2 4 h after surgery and on postoperative days 2, 3 and 7. On each occasion inspired oxygen fraction was changed in four to six steps to obtain arterial oxygen saturation (SaO2) in the range of 90-100%, enabling construction of FEO2/SaO2 curves. Measurements of ventilation, circulation and oxygenation were entered in a previously described mathematical model of pulmonary gas exchange. RESULTS: We found that oxygenation was most impaired 3 days after surgery. By fitting the mathematical model to the FEO2/SaO2 curve, we found that shunt remained constant throughout the study period. However, deltaPO2 increased from 0.5 kPa (median, range 0-3.8) 2-4 h after surgery, to 3.2 kPa (range 1.2-6.4, P < 0.05) on day 2, and to 4.0 kPa (range 1.2-8.3) on day 3. On day 7, deltaPO2 decreased to 2.2 kPa (range 0-3.5, P < 0.05). CONCLUSIONS: Ventilation/perfusion mismatch (deltaPO2), rather than shunt, explains the changes in postoperative oxygenation. The model of pulmonary gas exchange may serve as a useful and potentially non-invasive clinical tool for monitoring patients at risk of postoperative hypoxaemia. PMID- 15109194 TI - Changes in respiratory physiological dead space and compliance during non abdominal, upper abdominal and lower abdominal surgery under general anaesthesia. AB - BACKGROUND AND OBJECTIVE: To evaluate the temporal changes in respiratory physiological dead space and dynamic compliance of the respiratory system during non-abdominal, upper abdominal and lower abdominal surgery under general anaesthesia with intermittent positive pressure ventilation. METHODS: Thirty-four adult patients were studied (non-abdominal surgery, n = 8; upper abdominal surgery, n = 13 and lower abdominal surgery in lithotomy position, n = 13). Physiological dead space was measured using the single breath carbon dioxide test. The physiological dead space to tidal volume ratio (VD/VT), dynamic compliance of respiratory system, expiratory tidal volume and respiratory rate were measured 10 min after tracheal intubation, and 30, 60 and 120 min later. RESULTS: In lower abdominal surgery group, VD/VT was significantly increased at 120 min compared with 0 min (P = 0.005) and 30 min (P = 0.009). There were no significant differences in VD/VT between the three groups at any time point. Compliance decreased significantly in patients with upper abdominal (120 min) and lower abdominal surgery (60 and 120 min), but there were no significant changes during non-abdominal surgery. CONCLUSIONS: We found that the VD/VT increased in patients undergoing lower abdominal surgery in lithotomy and head down tilt, and compliance decreased in those undergoing upper abdominal and lower abdominal surgery over time. PMID- 15109195 TI - Differential platelet receptor expression following hydroxyethyl starch infusion in thrombocytopaenic orthotopic liver transplantation recipients. AB - BACKGROUND AND OBJECTIVE: Platelet function abnormalities influence the haemostatic defect in patients with liver failure. Patients after orthotopic liver transplantation present thrombocytopaenia associated with bleeding problems, which may be aggravated by the interaction of hydroxyethyl starches with platelets. METHODS: From 12 patients after liver transplantation venous blood samples (3 mL) were taken before, 20 and 120 min after infusion of hydroxyethyl starch of medium molecular weight (200 kDa/0.5) 6% 10 mL kg(-1) over a period of 30 min. Surface expression of glycoprotein IIb/IIIa and P-selectin were quantified by flow cytometry as well as the percentage of platelet-leucocyte complexes. RESULTS: A significant decrease of P-selectin expression following administration of hydroxyethyl starch after 120 min (89.1 +/- 4.2%, P = 0.029) and a corresponding significant reduction in the formation of platelet-monocyte complexes (81.1 +/- 7.8%, P = 0.001) were observed. There was no alteration in the glycoprotein IIb/IIIa expression after hydroxyethyl starch infusion. CONCLUSIONS: Infusion of hydroxyethyl starch 200 kDa/0.5 in clinically relevant doses does not alter glycoprotein IIb/IIIa expression in thrombocytopaenic patients with pre-existing platelet dysfunction after orthotopic liver transplantation. Accordingly, infusion of hydroxyethyl starch may have a beneficial effect on microvascular graft perfusion through the resulting haemodilution and reduced P-selectin expression with subsequent reduced leucocyte platelet complexes and endothelial adhesion. PMID- 15109196 TI - Ethnic differences in propofol and fentanyl response: a comparison among Caucasians, Kenyan Africans and Brazilians. AB - BACKGROUND AND OBJECTIVE: Differences in sensitivity to anaesthetic drugs may exist among human races. Allelic variants for drug metabolizing isoenzymes and other pharmacokinetic/pharmacodynamic differences may account for a variable response to anaesthetic drugs. This study was designed to investigate comparatively the anaesthetic requirements and the recovery trends of three different ethnic groups: Caucasians, African blacks and Brazilians. METHODS: The anaesthetic depth and recovery of groups of 45 patients undergoing total intravenous anaesthesia with propofol and fentanyl were compared. The bispectral index and clinical parameters were used to assess the depth of anaesthesia. The bispectral index, the response to verbal stimuli and the eye opening time were used to assess recovery. RESULTS: After stopping propofol, the bispectral index values of Caucasians returned to the baseline in about 10.8 +/- 4 min, that of Kenyan African blacks in 18 +/- 7 min and that of Brazilians in a highly variable time ranging from 5 to 25 min, (14.9 +/- 9.9). The time from discontinuation of propofol and fentanyl infusion to eye opening was 18.8 +/- 7.1 min in African blacks (P < 0.01) and 13.5 +/- 8.8 min in Brazilians (P > 0.05) vs. 11.6 +/- 4.5 min in Caucasians. Time to respond to verbal commands was 16.8 +/- 8 min in African blacks (P < 0.01) and 12.8 +/- 8.1 min in Brazilians (P > 0.05) vs. 9.9 +/- 4.5 min in Caucasians. CONCLUSIONS: The recovery of Kenyan African blacks from anaesthesia with propofol and fentanyl is much slower, in comparison with Caucasians. The recovery time of Brazilians is much more variable, in comparison with Caucasians. PMID- 15109197 TI - Patient safety during anaesthesia for magnetic resonance imaging. PMID- 15109198 TI - In off-pump cardiac surgery, neuromuscular monitoring should be mandatory and immediate extubation is feasible. PMID- 15109199 TI - Spurious bispectral index values due to electromyographic activity. PMID- 15109200 TI - Successful sequential double-lung transplantation for adult respiratory distress syndrome after long-term mechanical ventilation. PMID- 15109201 TI - Severe postoperative bleeding due to acquired haemophilia. PMID- 15109202 TI - Historical data on the neuraxial administration of opioids. PMID- 15109203 TI - Bispectral index monitoring in patients undergoing cardiac surgery under cardiopulmonary bypass: the influence of temperature changes should be considered. PMID- 15109204 TI - Potential neuroprotective properties of atracurium and cisatracurium in neurosurgical anaesthesia. PMID- 15109205 TI - Computational physiology and the Physiome Project. AB - Bioengineering analyses of physiological systems use the computational solution of physical conservation laws on anatomically detailed geometric models to understand the physiological function of intact organs in terms of the properties and behaviour of the cells and tissues within the organ. By linking behaviour in a quantitative, mathematically defined sense across multiple scales of biological organization--from proteins to cells, tissues, organs and organ systems--these methods have the potential to link patient-specific knowledge at the two ends of these spatial scales. A genetic profile linked to cardiac ion channel mutations, for example, can be interpreted in relation to body surface ECG measurements via a mathematical model of the heart and torso, which includes the spatial distribution of cardiac ion channels throughout the myocardium and the individual kinetics for each of the approximately 50 types of ion channel, exchanger or pump known to be present in the heart. Similarly, linking molecular defects such as mutations of chloride ion channels in lung epithelial cells to the integrated function of the intact lung requires models that include the detailed anatomy of the lungs, the physics of air flow, blood flow and gas exchange, together with the large deformation mechanics of breathing. Organizing this large body of knowledge into a coherent framework for modelling requires the development of ontologies, markup languages for encoding models, and web-accessible distributed databases. In this article we review the state of the field at all the relevant levels, and the tools that are being developed to tackle such complexity. Integrative physiology is central to the interpretation of genomic and proteomic data, and is becoming a highly quantitative, computer-intensive discipline. PMID- 15109206 TI - A definition of internal constancy and homeostasis in the context of non equilibrium thermodynamics. AB - The constancy of the internal environment, internal homeostasis, and its stability are necessary conditions for the survival of a biological system within its environment. These have never been clearly defined. For this purpose nonequilibrium thermodynamics is taken as a reference, and the essential principles of equilibrium, reversibility, stationary steady state and stability (Lyapounov, asymptotic, local and global), are briefly illustrated. On this basis, internal homeostasis describes a stationary state of nonequilibrium, the actual state of rest, X(t), resulting from the relation X(t) = Xs + x(t), between a time-independent steady state of reference (Xs), and time-dependent fluctuations of the state variables, x(t). In humans, two resting spontaneous homeostatic states are: (1) the conscious state of quiet wakefulness, during which time-dependent variables display bounded oscillations around the mean time independent steady state level, this conscious state being thus stable in the sense of Lyapounov, and (2) the unconscious stable state of non-rapid eye movement sleep, in which the time-dependent variables would approach the lowest spontaneously attainable time-independent state asymptotically, sleep becoming a globally stable and attractive state. Exercise may be described as a non-resting, unstable active state far away from equilibrium and hibernation is a resting, time-independent steady state very near equilibrium. The range between sleep and exercise is neurohumorally regulated. For spontaneously stable states to occur, slowing of the metabolic rate, withdrawal of the sympathetic drive and reinforcement of the vagal tone to the heart and circulation are required, thus confirming that the parasympathetic division of the autonomic nervous system is the main controller of homeostasis. PMID- 15109207 TI - Retrospective view of the carotid body research of Ronan G. O'Regan. AB - The Irish experimental physiologist Ronan G. O'Regan has made a substantial contribution to the field of autonomic control of the carotid body. This brief retrospective has been written by one of his former students, one of many to whom the beauty of Nature's design has been revealed through O'Regan's lectures and scientific papers. I shall describe some of his experiments that were performed from 1965 to 1981 in this historical sketch. PMID- 15109208 TI - Effects of chronic intermittent asphyxia on haematocrit, pulmonary arterial pressure and skeletal muscle structure in rats. AB - Sleep-disordered breathing in humans is a common condition associated with serious cardiovascular and other abnormalities. The prevalence and pathogenesis of increased haematocrit and pulmonary hypertension is controversial and it has been suggested that these changes only occur in patients who also have daytime continuous hypoxaemia. The hypothesis tested here is that the chronic intermittent hypoxia and asphyxia associated with sleep-disordered breathing causes erythropoiesis and pulmonary hypertension and that this occurs in the absence of periods of continuous hypoxia. In humans and animals with obstructive sleep apnoea, there are abnormalities of upper airway muscle structure that have been ascribed to increased load placed on these muscles. An alternative hypothesis is that chronic intermittent hypoxia and asphyxia cause changes in upper airway muscle structure and function. To test these hypotheses, rats were exposed to intermittent hypoxia and asphyxia for 8 h per day for 5 weeks. This caused an increase in haematocrit, right ventricular weight and pulmonary arterial pressure. There were only slight changes in diaphragm, upper airway and limb muscle structure and force production but in general, muscle fatigability was increased. In conclusion chronic intermittent hypoxia and asphyxia cause an increase in haematocrit and pulmonary arterial pressure in the absence of periods of continuous hypoxia. Chronic intermittent hypoxia and asphyxia have little effect on skeletal muscle structure and force production but increase muscle fatigue. Increased upper airway muscle fatigue could lead to a vicious cycle of further compromise in upper airway patency and further hypoxia and asphyxia. PMID- 15109209 TI - ATP is a key mediator of central and peripheral chemosensory transduction. AB - Recent evidence suggests that ATP is a mediator of central (within the ventral surface of the medulla) and peripheral (within the carotid body) chemosensory transduction. This short review discusses the data obtained in experiments in vivo and in vitro supporting this hypothesis. P2 receptors for ATP are expressed within the ventrolateral medulla as well as by the peripheral chemosensory afferent neurones. Blockade of P2 receptors in the ventrolateral medulla attenuates the CO2-induced increase in respiration while blockade of purinergic signalling impairs carotid body function and diminishes the ventilatory response to hypoxia. Furthermore, ATP is released from the ventral surface of the medulla during hypercapnia and from the carotid body during hypoxia. Finally, exogenous ATP applied on the ventral surface of the medulla evokes rapid increase in phrenic nerve activity, while ATP applied to the carotid body evokes marked excitation of the carotid sinus nerve afferents. We suggest that in the ventrolateral medulla ATP is produced following CO2/H(+)-induced activation of central chemosensory elements (neuronal and/or glial) and acts within the respiratory network to produce physiologically relevant changes in ventilation. In the carotid body, ATP contributes in a significant manner to the transmission of the sensitivity of the carotid body to changes in arterial PO2 and may be considered as a key transmitter released by chemoreceptor cells to activate endings of the sinus nerve afferent fibres. PMID- 15109210 TI - Interactions of chemostimuli at the single cell level: studies in a model system. AB - The responses of afferent chemosensory fibres of the carotid body to individual chemostimuli have long been established. However, the mechanisms underlying the multiplicative interactions of these stimuli (i.e. how the combined effects of hypoxia and hypercapnia exert a greater effect on afferent nerve discharge than the sum of their individual effects) have not been elucidated. Using the membrane hypothesis for carotid body chemoreception, in which chemostimuli inhibit type I cell K+ channels, leading to depolarization, voltage-gated Ca2+ entry and hence the triggering of exocytosis, this article considers data acquired in isolated type I carotid body cells and model chemoreceptor (PC12) cells to attempt to explain stimulus interactions. Whilst stimulus interactions are not clearly evident at the level of K+ channel inhibition or rises of [Ca2+]i, they are apparent at the level of transmitter release. Thus, it is clear that individual chemoreceptor cells can sense multiple stimuli, and that interactions of these stimuli can produce greater than additive effects in terms of transmitter release. PMID- 15109211 TI - Structural basis of hypoxic pulmonary hypertension: the modifying effect of chronic hypercapnia. AB - Exposure to chronic hypoxia causes pulmonary hypertension and pulmonary vascular remodelling. In chronic lung disease, chronic hypercapnia frequently coexists with hypoxia and is associated with worsening of pulmonary hypertension. It is generally stated that pulmonary hypertension in these conditions is secondary to hypoxic vascular remodelling and that hypercapnia augments this remodelling thus worsening the hypertension. We review recent evidence which shows that although chronic hypoxia causes thickening of the walls of pulmonary arterioles, these changes do not lead to structural narrowing of the lumen by encroachment. Moreover, hypoxia leads to new vessel formation within the pulmonary vasculature and not loss of vessels as formerly thought. Such neovascularization may provide a beneficial adaptation by increasing the area of the gas exchange membrane. These novel structural findings are supported by recent reports that inhibitors of the RhoA pathway can acutely reduce pulmonary vascular resistance in chronically hypoxic lungs to near normal values, demonstrating that structural changes are not the dominant mechanisms underling hypoxic pulmonary hypertension. Chronic hypercapnia inhibits the development of hypoxic pulmonary hypertension, pulmonary vascular remodelling and hypoxia-induced angiogenesis. This last effect might be maladaptive, as it would prevent the potentially beneficial increase in gas exchange membrane area. These findings suggest that structural narrowing of the vascular lumen of resistance vessels is not the mechanism by which hypoxia and hypercapnia cause pulmonary hypertension in chronic lung disease. PMID- 15109212 TI - Effect of polyphenolic compounds on the renal Na+,K(+)-ATPase during development and persistence of hypertension in rats. AB - It has been suggested that polyphenolic substances provide protection against the risk factors of cardiovascular diseases. The present study was designed to investigate whether application of red wine polyphenols influences the kinetic properties of the renal Na+,K(+)-ATPase in rats with hypertension (164 +/- 8 mmHg) that was experimentally induced by the NO synthase inhibitor N(G.) -nitro-L arginine methyl ester (L-NAME). Polyphenols in a dose of 40 mg kg(-1) day(-1) in drinking fluid induced different effects on the properties of the renal Na+,K(+) ATPase depending on the mode of their administration. Preventive application of polyphenols during the development of hypertension (144 +/- 5 mmHg) partially protected the Na+,K(+)-ATPase molecule against hypertension-induced deterioration via increased capability of the enzyme to bind ATP and/or Na+ as suggested by decrease of Km and KNa, respectively, even to values lower than in controls. However, polyphenols did not prevent the hypertension-induced reduction of the number of active Na+,K(+)-ATPase molecules as shown by similar V(max) values as compared to the hypertensive L-NAME group. The above protection is probably secured by a NO-dependent mechanism as suggested by 150% increase of the NO synthesis. Additional treatment of already hypertensive animals with polyphenols (153 +/- 8 mmHg) resulted in partial restoration of the Na+,K(+)-ATPase affinities especially for sodium as indicated by significant diminution of KNa. However, polyphenols in this mode of application did not slow down the L-NAME induced decrease in the number of Na+,K(+)-ATPase molecules in the kidney as suggested by additional significant decrease in V(max) values when comparing this group with the control group and also the hypertensive L-NAME group. In this case the polyphenols affected the Na,K-ATPase molecule in a NO-independent way as indicated by the fact that polyphenols failed to restore normal NO synthesis. PMID- 15109213 TI - Comparison of the firing patterns of human postganglionic sympathetic neurones and spinal alpha motoneurones during brief bursts. AB - Focal recordings from individual postganglionic sympathetic neurones in awake human subjects have revealed common firing properties. One of the most striking features is that they tend to fire only once per sympathetic burst. Why this should be so is not known, but we propose that the short duration of the burst may limit the number of times a sympathetic neurone can fire. Indeed, while the normal variation in cardiac interval and burst duration is too narrow to reveal a correlation between burst duration and the number of spikes generated, we know that spike generation is doubled when burst duration is doubled following ectopic heart beats. To test the hypothesis that the burst duration constrains the firing of individual sympathetic neurones to one per burst, we used the human skeletomotor system as a model for the sympathetic nervous system, which allowed us to vary burst duration and amplitude experimentally. Intramuscular recordings were made from 27 single motor units (alpha motoneurones) in the tibialis anterior or soleus muscles of seven subjects; multiunit EMG activity was recorded via surface electrodes and blood pressure was recorded continuously. Subjects were instructed to generate EMG bursts of varying amplitude in the intervals between heart beats. By constraining the firing of alpha motoneurones to brief ( approximately 400 ms) bursts we could emulate real sympathetic bursts. Individual motoneurones generated 0-7 spikes during the emulated sympathetic bursts, with firing patterns similar to those exhibited by real sympathetic neurones. Eleven motor units showed significant positive linear correlations between the number of spikes they generated within a burst and its amplitude, whereas for 17 motor units there were significant positive correlations between the number of spikes and burst duration. This indicates that burst duration is a major determinant of the number of times an alpha motoneurone will fire during a brief burst, and we suggest that the same principle may explain the firing pattern typical of human sympathetic neurones. PMID- 15109214 TI - Clenbuterol antagonizes glucocorticoid-induced atrophy and fibre type transformation in mice. AB - Beta-agonists and glucocorticoids are frequently coprescribed for chronic asthma treatment. In this study the effects of 4 week treatment with beta-agonist clenbuterol (CL) and glucocorticoid dexamethasone (DEX) on respiratory (diaphragm and parasternal) and limb (soleus and tibialis) muscles of the mouse were studied. Myosin heavy chain (MHC) distribution, fibres cross sectional area (CSA), glycolytic (phosphofructokinase, PFK; lactate dehydrogenase, LDH) and oxidative enzyme (citrate synthase, CS; cytochrome oxidase, COX) activities were determined. Muscle samples were obtained from four groups of adult C57/B16 mice: (1) Control (2) Mice receiving CL (CL, 1.5 mg kg(-1) day(-1) in drinking water) (3) Mice receiving DEX (DEX, 5.7 mg kg(-1) day(-1) s.c.) (4) Mice receiving both treatments (DEX + CL). As a general rule, CL and DEX showed opposite effects on CSA, MHC distribution, glycolytic and mitochondrial enzyme activities: CL alone stimulated a slow-to-fast transition of MHCs, an increase of PFK and LDH and an increase of muscle weight and fibre CSA; DEX produced an opposite (fast-to-slow transition) change of MHC distribution, a decrease of muscle weight and fibre CSA and in some case an increase of CS. The response varied from muscle to muscle with mixed muscles, as soleus and diaphragm, being more responsive than fast muscles, as tibialis and parasternal. In combined treatments (DEX + CL), the changes induced by DEX or CL alone were generally minimized: in soleus, however, the effects of CL predominated over those of DEX, whereas in diaphragm DEX prevailed over CL. Taken together the results suggest that CL might counteract the unwanted effects on skeletal muscles of chronic treatment with glucocorticoids. PMID- 15109215 TI - Gastric emptying and fluid availability after ingestion of glucose and soy protein hydrolysate solutions in man. AB - The double sampling gastric aspiration method was used to measure the effect of energy content on the rate of gastric emptying of glucose and soy protein hydrolysate solutions. The net rate of absorption of water from these solutions was assessed using deuterium oxide as a tracer for water. Six healthy male subjects were each studied on four separate occasions using a test drink volume of 600 ml. The half emptying times (t 1/2, median (range)) of the iso-energetic soy protein hydrolysate (6P, 60 g l(-1), 36 (14-39) min) and glucose (7G, 70 g l( 1), 25 (19-29) min) solutions were similar. These two solutions (6P, 7G) delivered energy to the small intestine at similar rates, and resulted in similar rates of accumulation of the deuterium tracer in the circulation. The dilute glucose solution (LG, 23 g l(-1)) was emptied faster (t 1/2 13 (11-19) min) and resulted in a faster rate of tracer accumulation in the circulation than any of the other solutions, including the iso-osmotic soy protein solution (LG 311 +/- 5 mosmol kg(-1), 6P 321 +/- 24 mosmol kg(-1)). The concentrated soy protein hydrolysate solution (12P, 120 g l(-1)) emptied more slowly (t 1/2 80 (44-120) min) than the more dilute solutions. The rate of energy delivery to the small intestine from 12P was similar to that from 6P for the first 50 min after ingestion, and similar to that from 7G at all sample points. These results indicate that the iso-energetic solutions of glucose and soy protein hydrolysate used in this study are emptied from the stomach at similar rates and result in similar rates of fluid availability after ingestion. PMID- 15109216 TI - Effect of time of day and rabbit strain on patterns of aortic wall permeability. AB - Lipid deposits occur more frequently downstream of branch points than upstream in immature rabbit and human aortas but the opposite pattern is seen in mature vessels. These distributions correlate spatially with age-related patterns of aortic permeability, observed in rabbits, and may be determined by them. The mature but not the immature pattern of permeability is dependent on endogenous nitric oxide synthesis. Although the transport patterns have hitherto seemed robust, recent studies have given the upstream pattern in some mature rabbits but the downstream pattern in others. Here we show that transport in mature rabbits is significantly skewed to the downstream pattern in the afternoon compared with the morning (P < 0.05), and switches from a downstream to an upstream pattern at around 21 months in rabbits of the Murex strain, but at twice this age in Highgate rabbits (P < 0.001). The effect of time of day was not explained by changes in nitric oxide production, assessed from plasma levels of nitrate and nitrate, nor did it correlate with conduit artery tone, assessed from the shape of the peripheral pulse wave. The effect of strain could not be explained by variation in nitric oxide production nor by differences in wall structure. The effects of time of day and rabbit strain on permeability patterns explain recent discrepancies, provide a useful tool for investigating underlying mechanisms and may have implications for human disease. PMID- 15109217 TI - Exercise and training effects on ceramide metabolism in human skeletal muscle. AB - In rat skeletal muscle prolonged exercise affects the content and composition of ceramides, but in human skeletal muscle no data are available on this compound. Our aim was to examine the content of ceramide- and sphingomyelin fatty acids and neutral, Mg(2+)-dependent sphingomyelinase activity in skeletal muscle in untrained and trained subjects before and after prolonged exercise. Healthy male subjects were recruited into an untrained (n = 8, VO2,max 3.8 +/- 0.2 1 min1) and a trained (n = 8, Vo2,max 5.1 +/- 0.1 1 min2) group. Before and after a 3-h exercise bout (58 +/- 1% VO2,max) a muscle biopsy was excised from the vastus lateralis. Ceramide and sphingomyelin were isolated using thin-layer chromatography. The content of individual ceramide fatty acids and sphingomyelin fatty acids was measured by means of gas-liquid chromatography. The activity of neutral, Mg(2+)-dependent sphingomyelinase was measured using N-[14CH3] sphingomyelin as a substrate. Prior to exercise, the muscle total ceramide fatty acid content in both groups was similar (201 +/- 18 and 197 +/- 9 nmol g(-1) in the untrained and trained group, respectively) and after exercise a 25% increase in the content was observed in each group. At rest, the muscle total sphingomyelin fatty acid content was higher in untrained than in trained subjects (456 +/- 10, 407 +/- 7 nmol g(-1), respectively; P < 0.05). After exercise a 20% increase (P < 0.05) in total sphingomyelin was observed only in the trained subjects. The muscle neutral, Mg(2+)-dependent sphingomyelinase activity was similar in the two groups at rest and a similar reduction was observed after exercise in both groups (untrained from 2.19 +/- 0.08 to 1.78 +/- 0.08 and trained from 2.31 +/- 0.12 to 1.80 +/- 0.09 nmol (mg protein) (-1) h(-1); P < 0.05 in each case). In conclusion, we have reported, for the first time, the values for ceramide fatty acid content and neutral, Mg2(+)-dependent sphingomyelinase activity in human skeletal muscle. The results indicate that acute prolonged exercise affects ceramide metabolism in human skeletal muscle both in untrained and in trained subjects and this may influence muscle cell adaptation and metabolism. PMID- 15109218 TI - Interaction between direct sympathetic and vagus nerve stimulation on heart rate in the isolated rabbit heart. AB - The interaction between the effects of vagus nerve stimulation (VS) and sympathetic stimulation (SS) on intrinsic heart rate was studied in the novel innervated isolated rabbit heart preparation. The effects of background VS, at different frequencies--2 Hz (low), 5 Hz (medium), 7 Hz (high)--on the chronotropic effects of different frequencies of SS--2 Hz (low), 5 Hz (medium), 10 Hz (high)--were studied. The experiments were repeated in the reverse direction studying the effects of different levels of background SS on the chronotropic effects of different levels of VS. Background VS reduced the overall positive chronotropic effect of SS at steady state in a frequency dependent manner and the rate of increase in heart rate during low and medium SS (but not high SS) was slowed in the presence of background VS. These results suggest that pre- and postjunctional mechanisms may be involved in the sympatho-vagal interaction on heart rate. On the other hand, the chronotropic effect of VS was enhanced in the presence of background SS. Vagal stimulation appears to play a dominant role over sympathetic stimulation in chronotropic effects on the isolated heart. The innervated isolated heart preparation is a valuable model to study the complex mechanisms underlying the interaction between sympathetic and parasympathetic stimulation on cardiac function. PMID- 15109219 TI - Retrograde transfer of steroid hormones to the ovary in luteal and follicular phases of porcine oestrous cycle in vivo. AB - The efficiency of the retrograde transfer of steroid ovarian hormones from the ovarian effluent into blood supplying the ovary and the rate of its back transport to the ovary were determined for the first time in in vivo conditions. Sexually mature gilts (n = 25) were used in the physiological study. The concentration of oestradiol and progesterone in blood collected from the ovarian artery was higher in both the follicular phase (by 87.9 +/- 2.9% and 150.0 +/- 4.8%, respectively, P < 0.001) and the luteal phase (by 82.1 +/- 3.9% and 77.7 +/ 2.7%, respectively, P < 0.001) than in systemic blood reaching the initial part of the ovarian artery. The high efficiency of the retrograde transfer was not dependent on the concentration of hormones in the ovarian venous blood. However, the efficiency and rate of the retrograde transfer differed between phases of the oestrous cycle. We suggest that such effective retrograde transfer of ovarian hormones must affect the secretory function of the ovary. PMID- 15109220 TI - Complication rates and patency of radiologically guided mushroom gastrostomy, balloon gastrostomy, and gastrojejunostomy: a review of 250 procedures. AB - PURPOSE: To compare complication rates and tube performance of percutaneous mushroom gastrostomy, balloon gastrostomy, and gastrojejunostomy. METHODS: Between September 9, 1999 and April 23, 2001, 203 patients underwent 250 radiologically guided percutaneous gastrostomy and gastrojejunostomy procedures. Follow-up was conducted through chart reviews and review of our interventional radiology database. Procedural and catheter-related complications were recorded. Chi-square statistical analysis was performed. RESULTS: In patients receiving mushroom-retained gastrostomy catheters (n = 114), the major complication rate was 0.88% (n = 1), the minor complication rate was 5.3% (n = 6), and the tube complication rate was 4.4% (n = 5). In patients receiving balloon-retained gastrostomy tubes (n = 67), the major complication rate was 0, the minor complication rate was 4.5% (n = 3), and the tube complication rate was 34.3% (n = 23). In patients receiving gastrojejunostomy catheters (n = 69), the major complication rate was 1.4% (n = 1), the minor complication rate was 2.9% (n = 2), and the tube complication rate was 34.8% (n = 24). No statistically significant differences were found between procedural or peri-procedural complications among the different types of tubes. Mushroom-retained catheters had significantly fewer tube complications (p < 0.01). CONCLUSIONS: Percutaneous gastrostomy and gastrojejunostomy have similar procedural and peri-procedural complication rates. Mushroom gastrostomy catheters have fewer tube-related complications compared with balloon gastrostomy and gastrojejunostomy catheters. In addition, mushroom retained catheters exhibit the best overall long-term tube patency and are therefore the gastrostomy catheter of choice. PMID- 15109221 TI - Multislice CT angiography in renal artery stent evaluation: prospective comparison with intra-arterial digital subtraction angiography. AB - PURPOSE: To assess the role of multislice computed tomography angiography (MCTA) in the evaluation of renal artery stents, using intra-arterial digital subtraction angiography (DSA) as the gold standard. METHODS: Twenty consecutive patients (15 men, 5 women) with 23 renal artery stents prospectively underwent both MCTA and DSA. Axial images, multiplanar reconstructions and maximum intensity projection images were used for diagnosis. The MCTA and DSA images were each interpreted without reference to the result of the other investigation. RESULTS: The three cases of restenosis on DSA were detected correctly by MCTA; in 19 cases where MCTA showed a fully patent stent, the DSA was also negative. Sensitivity and negative predictive value (NPV) of MCTA were therefore 100%. In four cases, MCTA showed apparently minimal disease which was not shown on DSA. These cases are taken as false positive giving a specificity of 80% and a positive predictive value of 43%. CONCLUSION: The high sensitivity and NPV suggest MCTA may be useful as a noninvasive screen for renal artery stent restenosis. MCTA detected mild disease in a few patients which was not confirmed on angiography. PMID- 15109222 TI - Use of double-occlusion balloon catheter: preoperative portal vein embolization for induction of future remnant liver hypertrophy. AB - PURPOSE: To evaluate the efficacy and safety of using double-occlusion balloon catheters in preoperative portal vein embolization (PVE) to induce future remnant liver hypertrophy. MATERIALS AND METHODS: PVE was achieved with gelatin sponges by using double-occlusion balloon catheter in seventeen patients with hepatobiliary malignant tumors. The ipsilateral approach was used in thirteen patients and the contralateral approach in four patients due to large size of tumor in the right hepatic lobe. Surgery was performed in 15 patients, 14-27 days (mean, 21.9 days) after PVE. Computed tomographic liver volumetric studies were performed before embolization and before surgery. The changes in aspartate aminotransferase (AST), alanine aminotransferase (ALT), prothrombin time (PT), and total bilirubin levels before and after the PVE were evaluated. Complications were evaluated after PVE. RESULTS: PVE using double-occlusion balloon catheter was successful in all cases, irrespective of approach technique. The future remnant liver (FRL) volumes were 251-920 cm3 (mean, 437 cm3) before PVE and 281 1042 cm3 (mean, 555 cm3) after PVE. The mean increase in the volume of the FRL was 28.6%; this represented 37% of the preresection volume of the liver. Clinical and biologic tolerance of PVE was mandatory. There were no complications. CONCLUSIONS: PVE using the double-occlusion balloon catheter is safe and well tolerated and can be performed technically with ease. This hypertrophy allows hepatectomy to be performed safely when the FRL volume is initially insufficient in patients with hepatobiliary tumors. PMID- 15109223 TI - Managing inadvertent arterial catheterization during central venous access procedures. AB - PURPOSE: Approximately 200,000 central venous catheterizations are carried out annually in the National Health Service in the United Kingdom. Inadvertent arterial puncture occurs in up to 3.7%. Significant morbidity and death has been reported. We report on our experience in the endovascular treatment of this iatrogenic complication. METHODS: Retrospective analysis was carried out of 9 cases referred for endovascular treatment of inadvertent arterial puncture during central venous catheterization over a 5 year period. RESULTS: It was not possible to obtain accurate figures on the numbers of central venous catheterizations carried out during the time period. Five patients were referred with carotid or subclavian pseudoaneurysms and hemothorax following inadvertent arterial catheter insertion and subsequent removal. These patients all underwent percutaneous balloon tamponade and/or stent-graft insertion. More recently 4 patients were referred with the catheter still in situ and were successfully treated with a percutaneous closure device. CONCLUSION: If inadvertent arterial catheterization during central venous access procedures is recognized and catheters removed, sequelae can be treated percutaneously. However, once the complication is recognized it is better to leave the catheter in situ and seal the artery percutaneously with a closure device. PMID- 15109224 TI - The outback catheter: a new device for true lumen re-entry after dissection during recanalization of arterial occlusions. AB - PURPOSE: To report the initial experience with a new catheter system (The Outback catheter) designed to allow fluoroscopically controlled re-entry of the true arterial lumen after subintimal guidewire passage during recanalization procedures of arterial occlusions. METHODS: The catheter was used in 10 patients with intermittent claudication caused by chronic segmental occlusions of the superficial femoral or popliteal arteries. In all patients, conventional guidewire recanalization had failed. RESULTS: In 8 patients, successful true lumen re-entry was achieved with the Outback catheter. Percutaneous transluminal angioplasty was successfully performed in these patients without complications. Two technical failures occurred in heavily calcified arteries. CONCLUSION: The Outback catheter was safe and effective when used in complicated recanalization procedures in the superficial femoral and popliteal artery and the tibial trunk. PMID- 15109225 TI - Complications of lower-extremity outpatient arteriography via low brachial artery. AB - We retrospectively evaluated low brachial artery puncture for arteriography and its complications as an alternative approach route for bilateral lower extremity run-off. Using the Seldinger technique and catheterization with a sheathless 4-F multiple side-hole pigtail catheter, we performed 2250 low brachial artery punctures in outpatients. The right brachial artery (RBA) was successfully punctured in 2039 patients; the left brachial artery (LBA) in 200. The transfemoral approach was used in 11 patients when catheterizing either of brachial arteries failed. Ten major or moderate complications (2 pseudoaneurysms, 2 thrombosis, 1 dissection and 5 hematomas) were encountered. Surgical intervention was necessary in three cases. There were no transient ischemic attacks. Twenty-one patients suffered temporary loss of radial pulse which returned spontaneously in less than 1 hour. One patient demonstrated prolonged loss of pulse which required heparin. Low brachial artery puncture and catheterization at the antecubital fossa is a very safe and cost-effective alternative to the femoral artery approach for lower extremity intra-arterial arteriography in the hands of experienced operators. The success rate in catheterizing one of the brachial arteries was 99.52% with a low significant complications rate of 0.44%. The transbrachial approach should be used as a standard method for lower extremity IA-DSA in an outpatient setting. PMID- 15109226 TI - Comparison of single-stick and double-stick techniques for percutaneous nephrostomy. AB - We compared single- and double-stick techniques of percutaneous nephrostomy insertion by retrospectively reviewing 140 percutaneous nephrostomy procedures in 101 patients. All procedures were performed by residents or fellows with direct attending supervision. Either the single-stick or double-stick technique was used based solely on personal attending preference. There were no significant differences in groups in terms of age, sex, or degree of hydronephrosis. In the single-stick technique, the kidney was punctured with sonographic guidance and the tract was serially dilated to accept an 8.5 Fr. nephrostomy catheter. In the double-stick technique, the kidney was punctured with sonographic guidance and a mixture of air and contrast were injected into the collecting system. The affected side was then elevated and a posterior calyx was punctured using fluoroscopic guidance. Both groups were compared in terms of complications and early tube dysfunction using the chi-squared test. All procedures were successful without immediate complications. Bleeding requiring transfusion occurred in 4.7% (4/86) procedures in the single stick group and 3.7% (2/54) in the double stick group (p-value not significant). None of these patients required further interventions for bleeding. Tube dysfunction leading to premature tube exchange occurred in 3.5% (3/86) of catheters in the single stick group and 3.7% (2/54) of catheters in the double-stick group (p-value not significant). We found no significant difference between the single and double-stick methods of percutaneous nephrostomy in terms of success rates, complications, or tube function. We believe that the single-stick method should be adopted as the insertion technique of choice. PMID- 15109227 TI - Intravascular ultrasound guidance for transjugular intrahepatic portosystemic shunt procedure in a swine model. AB - A new method is described for guiding hepato-portal venous puncture using a longitudinal side-view intravascular ultrasound (L-IVUS) transducer to assist in the performance of transjugular intrahepatic portosystemic shunt (TIPS) in three Australian swine. Simultaneous L-IVUS with an AcuNav 5-10 MHz 10 Fr transducer (Acuson Corporation, Mountain View, CA, USA) and fluoroscopy guidance was used to image and monitor the hepatic to portal venous puncture, dilatation of the tract, and deployment of the TIPS stent. Flow through the shunt could be demonstrated with both L-IVUS and angiography. TIPS was successful in all swine. The time for portal vein puncture once the target portal vein was identified was reduced at each attempt. The number of portal vein puncture attempts was 2, 1, and 1. No post-procedural complication was evident. L-IVUS-guided TIPS is practical and has the potential to improve safety by permitting simultaneous ultrasound and fluoroscopic imaging of the needle and target vascular structures. This technique allows for a more streamlined approach to TIPS, decreasing the fluoroscopic time (hence, decreasing the radiation exposure to the staff and patient) and anesthetic time. In addition, there are improved safety benefits obviating the need for wedged portography, facilitating avoidance of bile duct and hepatic arterial puncture, and minimizing hepatic injury by decreasing liver capsular puncture and the attendant risks. PMID- 15109228 TI - Post-dilatation intravascular brachytherapy trials on hypercholesterolemic rabbits using 32P-phosphate solutions in angioplasty balloons. AB - PURPOSE: Response of peripheral arteries to post-dilatation intravascular brachytherapy (IVBT) using 32P liquid sources was studied in a rabbit model. METHODS: The applied sources were angioplasty balloons filled with aqueous solutions of Na2H32PO4, NaCl and iodinated contrast. Dose distribution was calibrated by thermoluminescence dosimetry. The uncertainty of in vitro determinations of the activity-dose dependence was +/- 15-30%. The animal experiments were performed on rabbits with induced hypercholesterolemia. The 32P sources were introduced into a randomly chosen (left or right) iliac artery, immediately after balloon injury. Due to the low specific activity of the applied sources, the estimated 7-49 Gy doses on the internal artery surface required 30 100 min irradiations. A symmetric, balloon-occluded but non-irradiated artery of the same animal served as control. Radiation effects were evaluated by comparing the thicknesses of various components of irradiated versus untreated artery walls of each animal. RESULTS: The treatment was well tolerated by the animals. The effects of various dose ranges could be distinguished although differences in individual biological reactions were large. Only the 49 Gy dose at "zero" distance (16 Gy at 1.0 mm from the balloon surface) reduced hypertrophy in every active layer of the artery wall. The cross-sectional intimal thicknesses after 7, 12, 38 and 49 Gy doses were 0.277, 0.219, 0.357 and 0.196 mm2 respectively, versus 0.114, 0.155, 0.421 and 0.256 mm2 in controls (p < 0.05). The lowest radiation dose on the intima induced the opposite effect. Edge intimal hyperplasia was not avoided, which agrees with other reports. The edge restenosis and the variability of individual response to identical treatment conditions must be considered as limitations of the post-dilatation IVBT method. CONCLUSION: Only application of highest irradiation doses was effective. The irradiation dose should be planned and calculated for adventitia. PMID- 15109229 TI - Angioplasty or stenting of extra- and intracranial vertebral artery stenoses. AB - PURPOSE: To determine the feasibility and safety of angioplasty or angioplasty and stenting of extra- and intracranial vertebral artery (VA) stenosis. METHODS: In 16 consecutive patients (9 men, 7 women; mean age 61 years, range 49-74 years) 16 stenotic VAs were treated with angioplasty or angioplasty and stenting. Eleven stenoses were localized in V1 segment, 1 stenosis in V2 segment and 4 stenoses in V4 segment of VA. Fourteen VA stenoses were symptomatic, 2 asymptomatic. The etiology of the stenoses was atherosclerotic in all cases. RESULTS: Angioplasty was performed in 8 of 11 V1 and 2 of 4 V4 segments of the VA. In 3 of 11 V1 segments and 2 of 4 V4 segments of the VA we combined angioplasty with stenting. The procedures were successfully performed in 14 of 16 VAs (87%). Complications were asymptomatic vessel dissection resulting in vessel occlusion in 1 of 11 V1 segments and asymptomatic vessel dissection in 2 of 4 V4 segments of the VA. One patient died in the 24-hr period after the procedure because of subarachnoid hemorrhage as a complication following vessel perforation of the treated V4 segment. CONCLUSION: Angioplasty or angioplasty and stenting of extracranial VA stenoses can be performed with a high technical success rate and a low complication rate. In intracranial VA stenosis the procedure is technically feasible but complications can be life-threatening. The durability and procedural complication rates of primary stenting without using predilation in extra- and intracranial VA stenosis should be defined in the future. PMID- 15109230 TI - Guenther tulip filter retrieval from a left-sided inferior vena cava. AB - Optional (retrievable) inferior cava filters (IVC) may have advantages over permanent filters in a certain subset of patients, especially in view of recent concerns about the long-term thrombotic complications of the latter. Retrieval of the Guenther Tulip Filter (GTF), an optional filter, has been reported in a total of 76 patients. We present the first description of GTF retrieval from a left sided IVC using the right internal jugular approach. PMID- 15109231 TI - Bird's nest filter causing symptomatic hydronephrosis following transmural penetration of the inferior vena cava. AB - We report a case of symptomatic hydronephrosis caused by transcaval penetration of a Bird's Nest filter. Perforation of the wall of the inferior vena cava (IVC) following insertion of a caval filter is a well-recognized complication. Whilst two cases of hydronephrosis have been described with Greenfield filters, no case involving a Bird's Nest filter has been reported previously. PMID- 15109232 TI - Application of balloon-occluded retrograde transvenous obliteration to gastric varices complicating refractory ascites. AB - We report two cases of gastric varices complicated by massive ascites that disappeared after balloon-occluded retrograde transvenous obliteration (B-RTO). The first patient had progressive gastric varices that continued to enlarge even after three episodes of esophagogastric variceal bleeding, and the second patient was admitted to our hospital because of the bleeding from gastric varices. After B-RTO procedures in both patients, significant improvement of the ascites, hepatic function reserve, and hypoalbuminemia was observed. Although further experience is needed, our experience points to the likelihood of the amelioration of ascites after B-RTO. PMID- 15109233 TI - Superior mesenteric artery stent-graft placement in a patient with pseudoaneurysm developing from a pancreatic pseudocyst. AB - Pseudoaneurysm is a relatively rare but serious complication of pancreatitis which is often fatal. We report successful stent-graft placement in the superior mesenteric artery in a 45-year-old man with a pancreatic pseudocyst that grew during therapy for chronic pancreatitis and developed into a pseudoaneurysm. After a stent graft was inserted in the superior mesenteric artery, the pseudoaneurysm disappeared and no further complications developed. Stentgraft placement was considered to be a useful therapy for pseudoaneurysms in the superior mesenteric arterial region. PMID- 15109234 TI - Endovascular treatment of an aortobronchial fistula. AB - A 67-year-old man operated on 8 years previously for type B aortic dissection presented with two episodes of massive hemoptysis. An aortobronchial fistula was suspected with spiral computed tomography angiography, and showed a small pseudoaneurysm corresponding to the distal anastomotic site. The patient underwent endovascular stent-graft implantation and is asymptomatic 8 months after the procedure. PMID- 15109235 TI - Treatment of a left internal mammary artery to pulmonary artery fistula with polytetrafluoroethylene covered stents. AB - Internal mammary artery (IMA) to pulmonary artery (PA) fistula is a rare complication of coronary artery bypass grafting (CABG) that may present as myocardial ischemia. We describe a case of left IMA-to-PA fistula treated with balloon expandable coronary polytetrafluoroethylene (PTFE) graft stents and review previously reported cases of this entity. PMID- 15109236 TI - Impaired pancreatic endocrine and exocrine responses in growth-retarded piglets. AB - The alteration of pancreatic endocrine and exocrine secretory responses induced by secretagogues and neural input was investigated in post-weaning growth retarded (GR) piglets. Blood and pancreatic juice were collected from these animals (6-8-weeks old). Plasma insulin and pancreatic digestive enzymes induced by nutrients, drugs and vagal stimulation were measured biochemically. The pancreas was inspected by immunohistochemical analysis. In GR piglets, the plasma glucose and insulin concentrations at the resting state were very low, and the secretory response was also markedly reduced, with maximum inhibition of 90% by glucose administration and 83% by arginine administration. The insulin secretion was not increased by 2-deoxy-D-glucose administration in GR piglets. The pancreatic juice secretions induced by vagal stimulation and secretagogues in GR piglets were not different from those induced in the control piglets. However, amylase activity in the pancreatic juice and in the pancreas was significantly decreased in GR piglets, although trypsin and chymotrypsin activities were not different. In the immunohistochemical analysis, the numbers of islets and the staining degree for insulin antibody also declined in the pancreases of GR piglets. These results indicated the reduction of insulin and amylase secretions from the pancreas in GR piglets, suggesting that a dysfunction of pancreatic endocrine and exocrine secretion during growth after weaning may be an important factor in the induction of growth retardation in piglets. PMID- 15109237 TI - An in vitro study on spontaneous cervical contractility in the cow during oestrus and diestrus. AB - Cervical smooth muscle specimens were collected from slaughtered cows in oestrus and diestrus. Longitudinal and circular specimens were mounted in organ baths and after equilibration time and 2 g pre-load, their physiologic isometric contractility was recorded for 2.5 h. Area under the curve (AUC), maximal amplitude (Amax), mean amplitude (Amen) and minimal amplitude (Amin) were studied. Differences between cycle phases, between muscle layers and over the recorded time periods were statistically evaluated using linear mixed-effect models. In the cow, physiologic contractility of the cervix decreased significantly over time for all variables evaluated (time as covariate on a continuous scale). Furthermore, for Amax, there was a tendency for an influence of muscle layer (longitudinal > circular). Contractility was not influenced by cycle phases. PMID- 15109238 TI - Nodal, uterine and meningeal gamma(delta) T-cell lymphomas in cattle. AB - Three cases of bovine gamma(delta) T-cell lymphoma without skin involvement are described. Case 1 was a 17-month-old Holstein heifer with generalized lymphadenopathy. Case 2 was a 4-year-old Holstein cow that had multiple tumour masses in the uterine body and horns. Case 3, a 23-month-old Holstein bull was presented with generalized tremor, nystagmus and hyperesthaesia, and there were several tumour masses in the meninges. Cases 1 and 2 had epitheliotropic neoplastic infiltrates in the tonsillar epithelium and endometrial glands, respectively. Immunohistochemistry showed CD3+, WC1+, CD79a- lymphoma cells in all cases, and perforin was positive in two cases. Electron-dense granules were present in many neoplastic cells of all cases. These findings supported the cytotoxic gamma(delta) T-cell origin of the present lymphomas. Bovine gamma(delta) T-cell lymphoma may originate in a wide variety of anatomical sites and may be classified into several histological subtypes. PMID- 15109239 TI - Cutaneous lymphangiosarcoma in a young dog: clinical, anatomopathological and lectinhistochemical description. AB - Lymphangiosarcoma is a rare tumour in domestic animals arising from lymphatic endothelial cells. Occasionally, microscopic differentiation with haemangiosarcoma may be difficult. The aim of the present study was to describe a lymphangiosarcoma in a 1-year-old female Doberman Pinscher dog and to characterize its lectinhistochemical binding pattern as compared with that of haemangiosarcoma. The dog was presented because of a cutaneous painful swelling located in the left axilla. Histological diagnosis confirmed lymphangiosarcoma. The dog was killed. Necropsy revealed mediastinal lymph nodes' involvement. Twenty lectins were tested in tissue sections of this case as well as in four haemangiosarcomas from other dogs. Staining intensity was issued upon optical density determinations. Percentage of lectinhistochemical staining area was also conducted. RCA-I showed the most intense and wide distributed labelling pattern for lymphangiosarcoma. PHA-E was the counterpart for haemangiosarcoma. Should similar results be obtained in further studies, such differences could aid in the differential diagnosis between lymphangiosarcoma and haemangiosarcoma when histological pictures were not conclusive. PMID- 15109240 TI - Transitional cell carcinoma of the renal pelvis in two dogs. AB - Transitional cell carcinoma (TCC) of the renal pelvis was found in two dogs, a 7 year-old male English Setter and a 11-year-old female Shetland shepherd. Affected dogs were presented for clinical examination without any specific symptoms but haematuria in case 1 and occurrence of whitish material in the urine of case 2; neoplastic disorders were discovered with ultrasonographic investigation and fine needle aspiration biopsy. Histopathological examination was carried out after nephrectomy and ureterectomy of the affected kidney of both dogs, and confirmed the diagnosis of non-invasive and low grade TCC in case 1 and of infiltrating TCC in case 2. The clinical, gross, cytological and histopathological features of these rare tumours originating from transitional epithelium of the renal pelvis are reported. PMID- 15109241 TI - Effects of the ACE inhibitor quinapril on echocardiographic variables in horses with mitral valve insufficiency. AB - Twenty horses with mitral valve insufficiency, but without signs of congestive heart failure, and five horses without signs of heart disease were examined before and after medication with an angiotensin-converting enzyme (ACE) inhibitor. The examination included echocardiography assessment as well as heart catheterization. The echocardiographic examination included B-mode, M-mode, conventional and colour Doppler techniques. For 8 weeks, all horses were treated with Accupro 20 (active substance: Quinapril) at an oral dose rate of 120 mg/horse/day. A follow-up of the horses with mitral valve insufficiency after 8 weeks revealed a statistically significant increase in the stroke volume and the cardiac output as well as a decrease in regurgitation velocity time integral (VTI). The regurgitation blood velocity remained the same. The severity of mitral valve insufficiencies revealed a moderate improvement in five horses, from moderate to mild, after therapy. Significant changes of cardiac dimension (B mode) and shortening fraction (M-mode) before and after treatment could not be observed. The owners' judgement of the horses' performance was that of a minor improvement. In the horses without clinical findings the results of examination before and after treatment remained the same. PMID- 15109242 TI - Reference intervals and age-related changes for platelet count, mean platelet volume and plateletcrit in healthy pre-weaning piglets in Italy. AB - Platelet count (PLT) mean platelet volume (MPV) and plateletcrit (PCT) were determined for 117 Landrace x Large White piglets aged 3-21 days; counts were performed with an automated blood cell counter (ABX Pentra 120). Reference values were estimated following the guidelines of the International Federation of Clinical Chemistry (IFCC) and the International Committee for Standardization in Haematology (ICSH). The calculated central 95% reference limits for PLT was 49.9 516.2 x 10(9)/1, for MPV 6.71-9.91 fl and for PCT 0.009-0.395%. When observations are divided into three age groups (about 1 week each) there is an increase in mean PLT count and PCT in 2-week piglets, and a decrease in MPV from the first to the third week of life. These reference values provide guidelines for interpreting for experimental and clinical observations, as well as for monitoring of the health status of similar aged piglets determined using automated impedance-light focusing methodology. PMID- 15109243 TI - Cervimetry and ultrasonographic observations of the cervix regression in dairy cows during the first 10 days post partum. AB - The aim of this study was to describe the involution process of the bovine cervix during the first 10 days post partum by using an intracervically introduced gauge and transrectal ultrasonography. Twenty-five pluriparous Holstein Friesian cows were included, which were randomized into three groups: group 1 (n = 10): transrectal ultrasonographic presentation of the formation of cervical folds; group 2 (n = 10): measurement of the reduction of the canalis cervicalis using an intracervically introduced gauge. The examinations were performed directly after expulsion and 2, 4, 6, 8, 12, 16, 18 and 24 h post partum, and in intervals of 24 h up to the tenth day. The involution process can be divided into four phases: 1: fast reduction up to the sixth hour; 2: protracted reduction up to the second day without constant formation of folds; 3: protracted reduction up to the seventh day with constant formation of folds; 4: re-opening of the canalis cervicalis maintaining the structure of the folds. The reduction process (phase 1- phase 3) can be shown as the sum of two exponential functions with different grading coefficients: y = reduction 1 x e(-b1 x t) + reduction 2 x e(-b2 x t). Reduction 1 is reduction in cm during the first phase and reduction 2 is reduction in cm during the second and the third phase. Group 3 (n = 5): to exclude an influence of the high frequency of examinations on the involution process the cows were examined immediately after parturition and on the fifth and tenth day postpartum using the same method as in groups 1 and 2. In conclusion, the involution process of the bovine cervix guarantees a fast protection of the cavum uteri and a drainage of the lochia at the end of the early puerperium. The re-opening phase after the seventh day has not yet been described in any other species and can be interpreted as the adaptation to the exsudative puerperium in the cow. PMID- 15109244 TI - Surgical treatment of gastric impaction in farmed ostriches. AB - Partial or complete impaction of the proventriculus and/or the ventriculus is usually seen in juvenile ostriches. Common impaction materials include stones, sand, hay stems and leaves, and even plastic and metallic objects. Six live ostriches showing signs of impaction and three dead ones suspected to have impacted stomachs were presented to the Department of Clinical Sciences of the Faculty of Veterinary Medicine of the Aristotle University of Thessaloniki. The birds manifested anorexia, whitish and decreased urine output, scant defecation, unwillingness to rise and consume water. Clinical examination revealed distended and firm proventriculi, while radiographs showed foreign objects in the proventriculi of all birds and the ventriculus of one of them. The living birds were successfully treated surgically and medically. Autopsy of the three dead ostriches revealed severe dehydration and impaction of the stomachs. Bacteriological and parasitological examination was negative. Amendments in management practices were important to minimize losses caused by impaction of the stomachs in ostriches, while proventriculotomy remains the most effective approach to the problem. PMID- 15109245 TI - Defective DNA repair as a potential mechanism for the rapid development of drug resistance in Plasmodium falciparum. AB - The development and spread of highly drug-resistant parasites pose a central problem in the control of malaria. Understanding mechanisms that regulate genomic stability, such as DNA repair, in drug-resistant parasites and during drug treatment may help determine whether this rapid onset of resistance is due to an increase in the rate at which resistance-causing mutations are generated. This is the first report to demonstrate DNA repair activities from the malaria-causing parasite Plasmodium falciparum that are specific for ultraviolet light-induced DNA damage. The efficiency of DNA repair differs dramatically among P. falciparum strains with varying drug sensitivities. Most notable is the markedly reduced level of repair in the highly drug-resistant W2 isolate, which has been shown to develop resistance to novel drugs at an increased rate when compared to drug sensitive strains. Additionally, the antimalarial drug chloroquine and other quinoline-like compounds interfered with the DNA synthesis step of the repair process, most likely a result of direct binding to repair substrates. We propose that altered DNA repair, either through defective repair mechanisms or drug mediated inhibition, may contribute to the accelerated development of drug resistance in the parasite. PMID- 15109246 TI - Determination of the nature of the heme environment in nitrosyl indoleamine 2,3 dioxygenase using Multiple-scattering analyses of X-ray absorption fine structure. AB - Multiple-scattering analysis of X-ray absorption fine structure data on the NO adducts of indoleamine 2,3-dioxygenase (IDO) and analysis of X-ray absorption near-edge structure (XANES) have provided the first direct structural information about the iron center for this ubiquitous mammalian metalloprotein. The IDO(II)NO adduct, which is likely to play a physiological role in the immune system, differs from similar adducts such as Mb(II)NO and Lb(II)NO in that the Fe-His bond is essentially broken. At 10 K, the Fe-N(p)(av) bond length = 2.00(2) A, Fe NO bond length = 1.75 A, and angle = 140 degrees, which are typical of five coordinate Fe(II)NO species. The XANES is also closer to that of five-coordinate model complexes than six-coordinate species. In addition to the Fe(II)NO species, there was a minor component of the Fe(III)NO adduct because of incomplete reduction of the Fe(II) species. This was also a five-coordinate center and consists of a linear Fe(II)NO(+) moiety with the Fe-N(p)(av) bond length = 2.00(2) A, Fe-NO bond length = 1.63(3) A, and angle = 179 degrees. The results indicate that both the blocking of the heme site to O(2) binding and conformational changes induced by breaking the Fe-N(epsilon) bond may be important mechanisms by which NO inhibits IDO in vitro and in vivo. PMID- 15109247 TI - An intersubunit disulfide bond prevents in vitro aggregation of a superoxide dismutase-1 mutant linked to familial amytrophic lateral sclerosis. AB - Familial amyotrophic lateral sclerosis (FALS) is linked to over 90 point mutations in superoxide dismutase-1 (SOD1), a dimeric metalloenzyme. The postmortem FALS brain is characterized by SOD1 inclusions in the motor neurons of regions in which neuronal loss is most significant. These findings, together with animal modeling studies, suggest that aggregation of mutant SOD1 produces a pathogenic species. We demonstrate here that a mutant form of SOD1 (A4V) that is linked to a particularly aggressive form of FALS aggregates in vitro, while wild type SOD1 (WT) is stable. Some A4V aggregates resemble amyloid pores formed by other disease-associated proteins. The WT dimer is significantly more stable than the A4V dimer, suggesting that dimer dissociation may be the required first step of aggregation. To test this hypothesis, an intersubunit disulfide bond between symmetry-related residues at the A4V dimer interface was introduced. The resultant disulfide bond (V148C-V148C') eliminated the concentration-dependent loss of enzymatic activity of A4V, stabilized the A4V dimer, and completely abolished aggregation. A drug-like molecule that could stabilize the A4V dimer could slow the onset and progression of FALS. PMID- 15109248 TI - Identification of novel inhibitors of the SARS coronavirus main protease 3CLpro. AB - SARS (severe acute respiratory syndrome) is caused by a newly discovered coronavirus. A key enzyme for the maturation of this virus and, therefore, a target for drug development is the main protease 3CL(pro) (also termed SARS-CoV 3CL(pro)). We have cloned and expressed in Escherichia coli the full-length SARS CoV 3CL(pro) as well as a truncated form containing only the catalytic domains. The recombinant proteins have been characterized enzymatically using a fluorescently labeled substrate; their structural stability in solution has been determined by differential scanning calorimetry, and novel inhibitors have been discovered. Expression of the catalytic region alone yields a protein with a reduced catalytic efficiency consistent with the proposed regulatory role of the alpha-helical domain. Differential scanning calorimetry indicates that the alpha helical domain does not contribute to the structural stability of the catalytic domains. Analysis of the active site cavity reveals the presence of subsites that can be targeted with specific chemical functionalities. In particular, a cluster of serine residues (Ser139, Ser144, and Ser147) was identified near the active site cavity and was susceptible to being targeted by compounds containing boronic acid. This cluster is highly conserved in similar proteases from other coronaviruses, defining an attractive target for drug development. It was found that bifunctional aryl boronic acid compounds were particularly effective at inhibiting the protease, with inhibition constants as strong as 40 nM. Isothermal titration microcalorimetric experiments indicate that these inhibitors bind reversibly to 3CL(pro) in an enthalpically favorable fashion, implying that they establish strong interactions with the protease molecule, thus defining attractive molecular scaffolds for further optimization. PMID- 15109249 TI - Ricin A-chain substrate specificity in RNA, DNA, and hybrid stem-loop structures. AB - Ricin toxin A-chain (RTA) is the catalytic subunit of ricin, a heterodimeric toxin from castor beans. Its ribosomal inactivating activity arises from depurination of a single adenine from position A(4324) in a GAGA tetraloop from 28S ribosomal RNA. Minimal substrate requirements are the GAGA tetraloop and stem of two or more base pairs. Depurination activity also occurs on stem-loop DNA with the same sequence, but with the k(cat) reduced 200-fold. Systematic variation of RNA 5'-G(1)C(2)G(3)C(4)[G(5)A(6)G(7)A(8)]G(9)C(10)G(11)C(12)-3' 12mers via replacement of each nucleotide in the tetraloop with a deoxynucleotide showed a 16-fold increase in k(cat) for A(6) --> dA(6) but reduced k(cat) up to 300-fold for the other sites. Methylation of individual 2'-hydroxyls in a similar experiment reduced k(cat) by as much as 3 x 10(-3)-fold. In stem-loop DNA, replacement of d[G(5)A(6)G(7)A(8)] with individual ribonucleotides resulted in small kinetic changes, except for the dA(6) --> A(6) replacement for which k(cat) decreased 6-fold. Insertion of d[G(5)A(6)G(7)A(8)] into an RNA stem-loop or G(5)A(6)G(7)A(8) into a DNA stem-loop reduced k(cat) by 30- and 5-fold, respectively. Multiple substitutions of deoxyribonucleotides into RNA stem-loops in one case (dG(5),dG(7)) decreased k(cat)/K(m) by 10(5)-fold, while a second change (dG(5),dA(8)) decreased k(cat) by 100-fold. Mapping these interactions on the structure of GAGA stem-loop RNA suggests that all the loop 2'-hydroxyl groups play a significant role in the action of ricin A-chain. Improved binding of RNA DNA stem-loop hybrids provides a scaffold for inhibitor design. Replacing the adenosine of the RTA depurination site with deoxyadenosine in a small RNA stem loop increased k(cat) 20-fold to 1660 min(-1), a value similar to RTA's k(cat) on intact ribosomes. PMID- 15109250 TI - Inhibition of ricin A-chain with pyrrolidine mimics of the oxacarbenium ion transition state. AB - Ricin A-chain (RTA) catalyzes the hydrolytic depurination of a specific adenosine at position 4324 of 28S rRNA. Kinetic isotope effects on the hydrolysis of a small 10mer stem-tetraloop oligonucleotide substrate established the mechanism of the reaction as D(N)*A(N), involving an oxacarbenium ion intermediate in a highly dissociative transition state. An inhibitor with a protonated 1,4-dideoxy-1,4 imino-D-ribitol moiety, a 4-azasugar mimic, at the depurination site in the tetraloop of a 14mer oligonucleotide with a 5 bp duplex stem structure had previously been shown to bind to RTA with a K(d) of 480 nM, which improved to 12 nM upon addition of adenine. Second-generation stem-tetraloop inhibitors have been synthesized that incorporate a methylene bridge between the nitrogen of a 1 azasugar mimic, namely, (3S,4R)-3-hydroxy-4-(hydroxymethyl)pyrrolidine, and substituents, including phenyl, 8-aza-9-deazaadenyl, and 9-deazaadenyl groups, that mimic the activated leaving group at the transition state. The values for the dissociation constants (K(i)) for these were 99 nM for the phenyl 10mer, 163 and 94 nM for the 8-aza-9-deazaadenyl 10- and 14mers, respectively, and 280 nM for the 9-deazaadenyl 14mer. All of these compounds are among the tightest binding molecules known for RTA. A related phenyl-substituted inhibitor with a deoxyguanosine on the 5'-side of the depurination site was also synthesized on the basis of stem-loop substrate specificity studies. This molecule binds with a K(i) of 26 nM and is the tightest binding "one-piece" inhibitor. 8-Aza-9-deaza- and 9-deazaadenyl substituents provide an increased pK(a) at N7, a protonation site en route to the transition state. The binding of these inhibitors is not improved relative to the binding of their phenyl counterpart, however, suggesting that RTA might also employ protonation at N1 and N3 of the adenine moiety to activate the substrate during catalysis. Studies with methylated adenines support this argument. That the various stem-loop inhibitors have similar potencies suggests that an optimal one-piece inhibitor remains to be identified. The second generation inhibitors described here incorporate ribose mimics missing the 2 hydroxy group. On the basis of inhibition data and substrate specificity studies, the 2'-hydroxyl group at the depurination site seems to be critical for recruitment as well as catalysis by RTA. PMID- 15109251 TI - Specificity of anion binding in the substrate pocket of bacteriorhodopsin. AB - The structure of the D85S mutant of bacteriorhodopsin with a nitrate anion bound in the Schiff base binding site and the structure of the anion-free protein have been obtained in the same crystal form. Together with the previously solved structures of this anion pump, in both the anion-free state and bromide-bound state, these new structures provide insight into how this mutant of bacteriorhodopsin is able to bind a variety of different anions in the same binding pocket. The structural analysis reveals that the main structural change that accommodates different anions is the repositioning of the polar side chain of S85. On the basis of these X-ray crystal structures, the prediction is then made that the D85S/D212N double mutant might bind similar anions and do so over a broader pH range than does the single mutant. Experimental comparison of the dissociation constants, K(d), for a variety of anions confirms this prediction and demonstrates, in addition, that the binding affinity is dramatically improved by the D212N substitution. PMID- 15109252 TI - Studies of the enzymic mechanism of Candida tenuis xylose reductase (AKR 2B5): X ray structure and catalytic reaction profile for the H113A mutant. AB - Xylose reductase from the yeast Candida tenuis (CtXR) is a family 2 member of the aldo-keto reductase (AKR) superfamily of proteins and enzymes. Active site His 113 is conserved among AKRs, but a unified mechanism of how it affects catalytic activity is outstanding. We have replaced His-113 by alanine using site-directed mutagenesis, determined a 2.2 A structure of H113A mutant bound to NADP(+), and compared catalytic reaction profiles of NADH-dependent reduction of different aldehydes catalyzed by the wild type and the mutant. Deuterium kinetic isotope effects (KIEs) on k(cat) and k(cat)/K(m xylose) show that, relative to the wild type, the hydride transfer rate constant (k(7) approximately 0.16 s(-1)) has decreased about 1000-fold in H113A whereas xylose binding was not strongly affected. No solvent isotope effect was seen on k(cat) and k(cat)/K(m xylose) for H113A, suggesting that proton transfer has not become rate-limiting as a result of the mutation. The pH profiles of log(k(cat)/K(m xylose)) for the wild type and H113A decreased above apparent pK(a) values of 8.85 and 7.63, respectively. The DeltapK(a) of -1.2 pH units likely reflects a proximally disruptive character of the mutation, affecting the position of Asp-50. A steady-state kinetic analysis for H113A-catalyzed reduction of a homologous series of meta-substituted benzaldehyde derivatives was carried out, and quantitative structure-reactivity correlations were used to factor the observed kinetic substituent effect on k(cat) and k(cat)/K(m aldehyde) into an electronic effect and bonding effects (which are lacking in the wild type). Using the Hammett sigma scale, electronic parameter coefficients (rho) of +0.64 (k(cat)) and +0.78 (k(cat)/K(m aldehyde)) were calculated and clearly differ from rho(k(cat)/K(aldehyde)) and rho(k(cat)) values of +1.67 and approximately 0.0, respectively, for the wild-type enzyme. Hydride transfer rate constants of H113A, calculated from kinetic parameters and KIE data, display a substituent dependence not seen in the corresponding wild type enzyme rate constants. An enzymic mechanism is proposed in which His-113, through a hydrogen bond from Nepsilon2 to aldehyde O1, assists in catalysis by optimizing the C=O bond charge separation and orbital alignment in the ternary complex. PMID- 15109253 TI - Effect of temperature on the structure of trout troponin C. AB - Adaptation for life at different temperatures can cause changes in many aspects of an organism. One example is the expression of different protein isoforms in species adapted to different temperatures. The calcium regulatory protein cardiac troponin C (cTnC), from rainbow trout (Oncorhynchus mykiss), is a good model for studying temperature effects, both because of its low physiological temperature and because mammalian cTnC, extensively studied at higher temperatures, can be used for comparison. We determined the structure and studied the backbone dynamics of the regulatory domain of trout cardiac troponin C (ScNTnC) with one Ca(2+) bound at 7 and 30 degrees C, using nuclear magnetic resonance spectroscopy (NMR). The overall fold of the regulatory domain of trout cTnC at both temperatures is similar to the regulatory domain of mammalian (human, bovine, and porcine isoform) cTnC bound to one Ca(2+). By comparing the trout structures at the two temperatures, we identify differences between the positions of the helices flanking the calcium binding loops, and the overall structure at 7 degrees C is more compact than that at 30 degrees C. The structure at 7 degrees C is more similar to the mammalian cTnC, which was determined at 30 degrees C, indicating that they have the same conformation at their respective physiological temperatures. The dynamic properties of the regulatory domain of trout cTnC are similar at the two temperatures that were used in these studies. PMID- 15109254 TI - Synaptobrevin transmembrane domain dimerization-revisited. AB - Synaptobrevin is a membrane-spanning soluble N-ethyl maleimid-sensitive factor (NSF) attachment protein receptor (SNARE) protein of synaptic vesicles that is essential for neurotransmitter release. Various lines of evidence indicate that it exists alternatively as a monomer, as a homodimer, as a heterodimer with synaptophysin, or as a ternary complex with other SNAREs at the various stages of the synaptic vesicle cycle. Homodimerization of synaptobrevin was previously shown by different authors to depend on its single transmembrane segment, and the crucial residues forming the helix-helix interface have been mapped. Since another recent study challenged these results, we reinvestigated this issue. Here, we show that native synaptobrevin can be cross-linked in synaptic vesicle membranes to a homodimer by disulfide bond formation between cysteine residues of the transmembrane segment. Further, we demonstrate that determination of synaptobrevin transmembrane segment interactions in membranes or in detergent solution requires careful control of experimental conditions. Thus, our present results corroborate that homodimerization of synaptobrevin is mediated by its transmembrane segment. PMID- 15109255 TI - Inflammatory cytokines and fatty acids regulate endothelial cell heparanase expression. AB - Heparan sulfates, the carbohydrate chains of heparan sulfate proteoglycans, play an important role in basement membrane organization and endothelial barrier function. We explored whether endothelial cells secrete a heparan sulfate degrading heparanase under inflammatory conditions and what pathways were responsible for heparanase expression. Heparanase mRNA and protein by Western blot were induced when cultured endothelial cells were treated with cytokines, oxidized low-density lipoprotein (LDL) or fatty acids. Heparanase protein in the cell media was induced 2-10-fold when cells were treated with tumor necrosis factor alpha (TNFalpha) or interleukin 1beta (IL-1beta). Vascular endothelial growth factor (VEGF), in contrast, decreased heparanase secretion. Inhibitors to nuclear factor-kappaB (NFkappaB), PI3-kinase, MAP kinase, or c-jun kinase (JNK) did not affect TNFalpha-induced heparanase secretion. Interestingly, inhibition of caspase-8 completely abolished heparanase secretion induced by TNFalpha. Fatty acids also induced heparanase, and this required an Sp1 site in the heparanase promoter. Immunohistochemical analyses of cross sections of aorta showed intense staining for heparanase in the endothelium of apoE-null mice but not wild-type mice. Thus, heparanase is an inducible inflammatory gene product that may play an important role in vascular biology. PMID- 15109256 TI - Ion binding affinity in the cavity of the KcsA potassium channel. AB - The hydrophobic cell membrane interior presents a large energy barrier for ions to permeate. Potassium channels reduce this barrier by creating a water-filled cavity at the middle of their ion conduction pore to allow ion hydration and by directing the C-terminal "end charge" of four alpha-helices toward the water filled cavity. Here we have studied the interaction of monovalent cations with the cavity of the KcsA K(+) channel using X-ray crystallography. In these studies, Tl(+) was used as an analogue for K(+) and the total ion-stabilization energy for Tl(+) in the cavity was estimated by measuring its binding affinity. Binding affinity for the Na(+) ion was also measured, revealing a weak selectivity ( approximately 7-fold) favoring Tl(+) over Na(+). The structures of the cavity containing Na(+), K(+), Tl(+), Rb(+), and Cs(+) are compared. These results are consistent with a fairly large (more negative than -100 mV) electrostatic potential inside the cavity, and they also imply the presence of a weak nonelectrostatic component to a cation's interaction with the cavity. PMID- 15109257 TI - Probing domain mobility in a flavocytochrome. AB - The crystal structures of various different members of the family of fumarate reductases and succinate dehydrogenases have allowed the identification of a mobile clamp (or capping) domain [e.g., Taylor, P., Pealing, S. L., Reid, G. A., Chapman, S. K., and Walkinshaw, M. D. (1999) Nat. Struct. Biol. 6, 1108-1112], which has been proposed to be involved in regulating accessibility of the active site to substrate. To investigate this, we have constructed the A251C:S430C double mutant form of the soluble flavocytochrome c(3) fumarate reductase from Shewanella frigidimarina, to introduce an interdomain disulfide bond between the FAD-binding and clamp domains of the enzyme, thus restricting relative mobility between the two. Here, we describe the kinetic and crystallographic analysis of this double mutant enzyme. The 1.6 A resolution crystal structure of the A251C:S430C enzyme under oxidizing conditions reveals the formation of a disulfide bond, while Ellman analysis confirms its presence in the enzyme in solution. Kinetic analyses with the enzyme in both the nonbridged (free thiol) and the disulfide-bridged states indicate a slight decrease in the rate of fumarate reduction when the disulfide bridge is present, while solvent-kinetic isotope studies indicate that in both wild-type and mutant enzymes the reaction is rate limited by proton and/or hydride transfer during catalysis. The limited effects of the inhibition of clamp domain mobility upon the catalytic reaction would indicate that such mobility is not essential for the regulation of substrate access or product release. PMID- 15109259 TI - Kinetic and thermodynamic analysis of the interaction of cations with dialkylglycine decarboxylase. AB - Dialkylglycine decarboxylase (DGD) is a tetrameric pyridoxal phosphate (PLP) dependent enzyme that catalyzes both decarboxylation and transamination in its normal catalytic cycle. Its activity is dependent on cations. Metal-free DGD and DGD complexes with seven monovalent cations (Li(+), Na(+), K(+), Rb(+), Cs(+), NH(4)(+), and Tl(+)) and three divalent cations (Mg(2+), Ca(2+), and Ba(2+)) have been studied. The catalytic rate constants for cation-bound enzyme (ck(cat) and ck(cat)/bK(AIB)) are cation-size-dependent, K(+) being the monovalent cation with the optimal size for catalytic activity. The divalent alkaline earth cations (Mg(2+), Ca(2+), and Ba(2+)) all give approximately 10-fold lower activity compared to monovalent alkali cations of similar ionic radius. The Michaelis constant for aminoisobutyrate (AIB) binding to DGD-PLP complexes with cations (bK(AIB)) varies with ionic radius. The larger cations (K(+), Rb(+), Cs(+), NH(4)(+), and Tl(+)) give smaller bK(AIB) ( approximately 4 mM), while smaller cations (Li(+), Na(+)) give larger values (approximately 10 mM). Cation size and charge dependence is also found with the dissociation constant for PLP binding to DGD-cation complexes (aK(PLP)). K(+) and Rb(+) possess the optimal ionic radius, giving the lowest values of aK(PLP). The divalent alkaline earth cations give aK(PLP) values approximately 10-fold higher than alkali cations of similar ionic radius. The cation dissociation constant for DGD-PLP-AIB-cation complexes (betaK(M)z+) was determined and also shown to be cation-size-dependent, K(+) and Rb(+) yielding the lowest values. The kinetics of PLP association and dissociation from metal-free DGD and its complexes with cations (Na(+), K(+), and Ba(2+)) were analyzed. All three cations tested increase PLP association and decrease PLP dissociation rate constants. Kinetic studies of cation binding show saturation kinetics for the association reaction. The half-life for association with saturating Rb(+) is approximately 24 s, while the half-life for dissociation of Rb(+) from the DGD-PLP-AIB-Rb(+) complex is approximately 12 min. PMID- 15109258 TI - Investigation of metal ion binding in phosphonoacetaldehyde hydrolase identifies sequence markers for metal-activated enzymes of the HAD enzyme superfamily. AB - The 2-haloalkanoic acid dehalogenase (HAD) family, which contains both carbon and phosphoryl transferases, is one of the largest known enzyme superfamilies. HAD members conserve an alpha,beta-core domain that frames the four-loop active-site platform. Each loop contributes one or more catalytic groups, which function in mediating the core chemistry (i.e., group transfer). In this paper, we provide evidence that the number of carboxylate residues on loop 4 and their positions (stations) on the loop are determinants, and therefore reliable sequence markers, for metal ion activation among HAD family members. Using this predictor, we conclude that the vast majority of the HAD members utilize a metal cofactor. Analysis of the minimum requirements for metal cofactor binding was carried out using Mg(II)-activated Bacillus cereus phosphonoacetaldehyde hydrolase (phosphonatase) as an experimental model for metal-activated HAD members. Mg(II) binding occurs via ligation to the loop 1 Asp12 carboxylate and Thr14 backbone carbonyl and to the loop 4 Asp186 carboxylate. The loop 4 Asp190 forms a hydrogen bond to the Mg(II) water ligand. X-ray structure determination of the D12A mutant in the presence of the substrate phosphonoacetaldehyde showed that replacement of the loop 1 Asp, common to all HAD family members, with Ala shifts the position of Mg(II), thereby allowing innersphere coordination to Asp190 and causing a shift in the position of the substrate. Kinetic analysis of the loop 4 mutants showed that Asp186 is essential to cofactor binding while Asp190 simply enhances it. Within the phosphonatase subfamily, Asp186 is stringently conserved, while either position 185 or position 190 is used to position the second loop 4 Asp residue. Retention of a high level of catalytic activity in the G185D/D190G phosphonatase mutant demonstrated the plasticity of the metal binding loop, reflected in the variety of combinations in positioning of two or three Asp residues along the seven-residue motif of the 2700 potential HAD sequences that were examined. PMID- 15109260 TI - DNA binding of methyl-CpG-binding protein MeCP2 in human MCF7 cells. AB - MeCP2 has been identified as a chromatin-associated protein that recognizes MAR elements as well as methyl-CpGs. To characterize target sequences of MeCP2 in human cells, we employed two complementary methods. First, by use of a preparative chromatin immunoprecipitation protocol, we created from MCF7 cells a library enriched with sequences bound to MeCP2. A total of 154 representative clones were sequenced and analyzed. A large fraction of clones was found to be associated with retrotransposons, mostly with Alu repeats. A subgroup of four clones is derived from putative MARs; one clone is associated with a CpG island, and four clones contain alphoid repeats. Classical satellite DNAs II and III are not represented among clones, although they are heavily methylated. Second, using indirect immunofluorescence microscopy, we show that MeCP2 staining of human metaphase chromosomes has a dotted to knobby appearance with a reduced level of staining of centromeric regions of some chromosomes. On the other hand, an anti-5 methylcytosine antibody preferentially stained the juxtacentromeric regions of chromosomes 1, 9, and 16, which habor highly methylated, classical satellite DNAs, and methylated alphoid sequences in centromeric regions of several other chromosomes with reduced intensity. In interphase MCF7 cells, the distribution of MeCP2 exhibits a granular appearance throughout the nucleus. This distribution does not parallel that of methylated cytosine and heterochromatin. The selective binding behavior of MeCP2 revealed by these results (preference for murine major satellite DNA, Alu sequences, MARs, and CpG islands) is explained by its ability to recognize the sequence information (guanine bases) adjacent to CpG (TpG) as demonstrated in previous footprinting experiments. PMID- 15109262 TI - Sequential ordered fatty acid alpha oxidation and Delta9 desaturation are major determinants of lipid storage and utilization in differentiating adipocytes. AB - Herein, we exploit the power of global lipidomics to identify the critical role of peroxisomal processing of fatty acids in adipocyte lipid storage and metabolism. Remarkably, 3T3-L1 differentiating adipocytes rapidly acquired the ability to alpha oxidize unbranched fatty acids, which is manifested in the accumulation of odd chain length unbranched fatty acids in all major lipid classes. Moreover, in differentiating adipocytes, unsaturated odd chain length fatty acids in TAG molecular species contained exclusively Delta9 olefinic linkages. Unsaturated fatty acids (e.g., oleic and palmitoleic acids) were not subject to alpha oxidation, resulting in the absence of Delta8 unsaturated odd chain length fatty acids. This highly selective substrate utilization resulted in the obligatory sequential ordering of alpha oxidation prior to Delta9 desaturation. On the basis of these results, a putative type 2 peroxisomal localization sequence was identified at the N-terminus of mouse stearoyl-CoA desaturase I (SCD I) comprised of (30)KVKTVPLHL(38). Kinetic analysis demonstrated that the rate of alpha oxidation of exogenously administered [9,10 (3)H]palmitic acid increased 4-fold during differentiation. Similarly, quantitative PCR demonstrated a 4-fold increase in phytanoyl-CoA alpha hydroxylase (PAHX) and fatty acyl-CoA oxidase (FACO) mRNA levels during differentiation. Collectively, these results underscore the role of peroxisomal fatty acid processing as an important determinant of the metabolic fate of fatty acids in the differentiating adipocyte. PMID- 15109261 TI - Structural characterization of the fibrillar form of the yeast Saccharomyces cerevisiae prion Ure2p. AB - The protein Ure2 from the yeast Saccharomyces cerevisiae has prion properties. It assembles in vitro into long, straight, insoluble fibrils that are similar to amyloids in that they bind Congo Red and show green-yellow birefringence and have an increased resistance to proteolysis. We recently showed that Ure2p fibrils assembled under physiologically relevant conditions are devoid of a cross-beta core. A model for fibril formation, where assembly is driven by non-native inter- and/or intramolecular interaction between Ure2p monomers following subtle conformational changes was proposed [Bousset et al. (2002) EMBO J. 21, 2903 2911]. An alternative model for the assembly of Ure2p into fibrils where assembly is driven by the stacking of 40-70 N-terminal amino acid residues of Ure2p into a central beta-core running along the fibrils from which the C-terminal domains protrude was proposed [Baxa et al. (2003) J. Biol. Chem. 278, 43717-43727]. We show here that Ure2p fibril congophilia and the associated yellow-green birefringence in polarized light are not indicative that the fibrils are of amyloid nature. We map the structures of the fibrillar and soluble forms of Ure2p using limited proteolysis and identify the reaction products by microsequencing and mass spectrometry. Finally, we demonstrate that the C-terminal domain of Ure2p is tightly involved in the fibrillar scaffold using a sedimentation assay and a variant Ure2p where a highly specific cleavage site between the N- and C terminal domains of the protein was engineered. Our results are inconsistent with the cross-beta-core model and support the model for Ure2p assembly driven by subtle conformational changes and underline the influence of the natural context of the N-terminal domain on the assembly of Ure2p. PMID- 15109263 TI - Conformational dynamics of the GdmHCl-induced molten globule state of creatine kinase monitored by hydrogen exchange and mass spectrometry. AB - Our understanding of the mechanism of protein folding can be improved by the characterization of folding intermediate states. Intrinsic tryptophan fluorescence measurements of equilibrium GdmHCl-induced unfolding of MM-CK allow for the construction of a "phase diagram", which shows the presence of five different conformational states, including three partially folded intermediates. However, only three states are detected by using pulsed-labeled H-D exchange analyzed by electrospray ionization mass spectrometry. One of them is the native state, and the two other species are present in proportions strongly dependent on the GdmHCl concentration and denaturation time. The low-mass peak is due to a largely exchange-incompetent state, which has gained only approximately 10 deuteriums more than the native protein. This population of MM-CK molecules has undergone a small conformational change induced by low GdmHCl concentrations. However, this limited change is in itself not sufficient to inactivate the enzyme or is easily reversible. The high-mass peak corresponds to a population of MM-CK that is fully deuterated. The comparison of fluorescence, activity, and H-D exchange measurements shows that the maximally populated intermediate at 0.8 M GdmHCl has the characteristics of a molten globule. It has no activity; it has 55% of its native alpha-helices and a maximum fluorescence emission wavelength of approximately 341 nm, and it binds ANS strongly. However, no protection against exchange is detected under the conditions used in this work. This paradox, the presence of significant residual secondary and tertiary structures detected by optical probes and the total deuteration of its amide protons detected by H-D exchange and mass spectrometry, could be explained by a highly dynamic MM-CK molten globule. PMID- 15109264 TI - A monomeric human apolipoprotein E carboxyl-terminal domain. AB - ApoE plays a critical role in lipoprotein metabolism and plasma lipid homeostasis through its high-affinity binding to the LDL-receptor family. In solution, apoE is an oligomeric protein and the C-terminal domain causes apoE's aggregation. The aggregation property presents a major difficulty for the structural determination of this protein. A high-level expression system of the apoE C-terminal domain is reported here. Using protein engineering techniques, we identified a monomeric, biologically active apoE C-terminal domain mutant. This mutant replaces five bulky hydrophobic residues in the region of residues 253-289 with either smaller hydrophobic or polar/charged residues (F257A, W264R, V269A, L279Q, and V287E). The solubility of the mutant is significantly increased ( approximately 10-fold). Cross-linking experiments indicate that this mutant is 100% monomeric even at 5 mg/mL. CD and guanidine hydrochloride denaturation results indicate that the mutant maintains an identical alpha-helical secondary structure and stability as compared with those of the wild-type protein. DMPC-binding assays demonstrate an identical vesicle clearance rate shared by both the mutant and the wild-type apoE C-terminal domain. In addition, electron microscopic results show identical recombinant HDL particles prepared with both the mutant and the wild-type proteins. These results indicate that residues F257, W264, V269, L279, and V287 are critical residues for aggregation but may not be important in maintaining the structure, stability, and lipid-binding activity of this apoE domain, suggesting that apoE may use different "epitopes" for its aggregation property, helical structure/stability, and lipid-binding activity. Finally, preliminary NMR data demonstrated that we have collected high-quality NMR spectra, allowing for an NMR structural determination of the apoE C-terminal domain. PMID- 15109265 TI - pH Dependence of heme iron coordination, hydrogen peroxide reactivity, and cyanide binding in cytochrome c peroxidase(H52K). AB - Replacement of the distal histidine, His-52, in cytochrome c peroxidase (CcP) with a lysine residue produces a mutant cytochrome c peroxidase, CcP(H52K), with spectral and kinetic properties significantly altered compared to those of the wild-type enzyme. Three spectroscopically distinct forms of the enzyme are observed between pH 4.0 and 8.0 with two additional forms, thought to be partially denatured forms, making contributions to the observed spectra at the pH extremes. CcP(H52K) exists in at least three, slowly interconverting conformational states over most of the pH range that was investigated. The side chain epsilon-amino group of Lys-52 has an apparent pK(a) of 6.4 +/- 0.2, and the protonation state of Lys-52 affects the spectral properties of the enzyme and the reactions with both hydrogen peroxide and HCN. In its unprotonated form, Lys-52 acts as a base catalyst facilitating the reactions of both hydrogen peroxide and HCN with CcP(H52K). The major form of CcP(H52K) reacts with hydrogen peroxide with a rate approximately 50 times slower than that of wild-type CcP but reacts with HCN approximately 3 times faster than does the wild-type enzyme. The major form of the mutant enzyme has a higher affinity for HCN than does native CcP. PMID- 15109266 TI - An apolipoprotein AI mimetic peptide: membrane interactions and the role of cholesterol. AB - The 18-amino acid amphipathic helical peptide Ac-DWFKAFYDKVAEKFKEAF-NH(2) promotes the separation of cholesterol from the phospholipid, resulting in the formation of cholesterol crystallites, even at mole fractions of cholesterol as low as 0.3. The peptide exerts a greater degree of penetration into membranes of pure phosphatidylcholine in the absence of cholesterol than into bilayers of phosphatidylcholine and cholesterol. The circular dichroism spectrum of the peptide in buffer indicates that it self-associates, leading to the formation of structures with higher helical content. However, in the presence of lipid, the peptide remains helical over a larger concentration range. The peptide undergoes a thermal transition on heating. Cholesterol has little effect on the secondary structure of the peptide; however, increased Trp emission intensity in the absence of cholesterol indicates a deeper penetration of the helix upon removal of cholesterol from the membrane. The results with these model systems demonstrate changes in peptide-lipid interactions that may be related to the observed biological properties of this peptide. PMID- 15109267 TI - Contribution of the hormone-response elements of the proximal ApoA-I promoter, ApoCIII enhancer, and C/EBP binding site of the proximal ApoA-I promoter to the hepatic and intestinal expression of the ApoA-I and ApoCIII genes in transgenic mice. AB - We have generated and studied the pattern of expression of transgenic mouse lines carrying the human apoA-I and apoCIII gene cluster mutated at different sites. In two lines, we have either mutated the hormone-response element (HRE) of element G of the apoCIII enhancer or the C/EBP binding site of the proximal apoA-I promoter. In a third line, we have mutated the two HREs of the apoA-I promoter and the HRE of the apoCIII enhancer. Mutations in the HRE of element G reduced the hepatic and intestinal expressions of the reporter chloramphenicol acetyltransferase (CAT) gene (which substituted the apoCIII gene) to 4 and 13% of the wild-type (WT) control, whereas the hepatic and intestinal expressions of the apoA-I gene were reduced to 92 and 25% of the WT control, respectively. A mutation in the C/EBP site increased the hepatic and intestinal expressions of the apoA-I gene approximately 1.25- and 1.6-fold, respectively, and did not affect the expression of the CAT gene. The mutation in the three HNF-4 binding sites of the apoA-I promoter/apoCIII enhancer nearly abolished the expression of apoA-I and the reporter CAT gene in all tissues. These findings establish the importance of the HREs for the hepatic and intestinal expressions of the apoA-I and apoCIII genes and suggest that C/EBP does not play a central role in the expression of the apoA-I gene. PMID- 15109268 TI - Contribution of factor VIIIa A2 and A3-C1-C2 subunits to the affinity for factor IXa in factor Xase. AB - Contributions of factor (F) VIIIa subunits to cofactor association with FIXa were evaluated. Steady-state fluorescence resonance energy transfer using an acrylodan labeled A3-C1-C2 subunit and fluorescein-Phe-Phe-Arg-FIXa yielded K(d) values of 52 +/- 10 and 197 +/- 55 nM in the presence and absence of phospholipid vesicles, respectively. A3-C1-C2 was an effective competitor of FVIIIa binding to FIXa as judged by inhibition of FXa generation performed in the absence of vesicles (K(i) approximately 1.6K(d) for FVIIIa-FIXa). However, the capacity for A3-C1-C2 to inhibit FVIIIa-dependent FXa generation in the presence of phospholipid was poor with a K(i) values (approximately 400 nM) that were approximately 100-fold greater than the K(d) for FVIIIa-FIXa interaction (4.2 +/- 0.6 nM). These results indicated that a significant component of the interprotein affinity is contributed by FVIIIa subunits other than A3-C1-C2 in the membrane-dependent complex. The isolated A2 subunit of FVIIIa interacts weakly with FIXa, and recent modeling studies have implicated a number of residues that potentially contact the FIXa protease domain (Bajaj et al. (2001) J. Biol. Chem. 276, 16302-16309). Site-directed mutagenesis of candidate residues in the A2 domain was performed, and recombinant proteins were stably expressed and purified. Functional affinity determinations demonstrated that one mutant, FVIII/Asp712Ala exhibited an 8-fold increased K(d) (35 +/- 1.5 nM) relative to wild-type suggesting a contribution by this residue of approximately 10% of the FVIIIa-FIXa binding energy. Thus both A2 and A3-C1-C2 subunits contribute to the affinity of FVIIIa for FIXa in the membrane-dependent FXase. PMID- 15109269 TI - Peptide repair of oxidative DNA damage. AB - Guanyl radical species are produced in DNA by electron removal caused by ionizing radiation, photoionization, oxidation, or photosensitization. DNA guanyl radicals can be reduced by electron donation from mild reducing agents. Important biologically relevant examples are the redox active amino acids cysteine, cystine, methionine, tryptophan, and tyrosine. We have quantified the reactivity of derivatives of these amino acids with guanyl radicals located in plasmid DNA. The radicals were produced by electron removal using the single electron oxidizing agent (SCN)(2)(*)(-). Disulfides (cystine) are unreactive. Thioethers (methionine), thiols (cysteine), and phenols (tyrosine) react with rate constants in the range 10(4)-10(6), 10(5)-10(6), and 10(5)-10(6) dm(3) mol(-1) s(-1), respectively. Indoles (tryptophan) are the most reactive with rate constants of 10(7)-10(8) dm(3) mol(-1) s(-1). Selenium analogues of amino acids are over an order of magnitude more reactive than their sulfur equivalents. Increasing positive charge is associated with a ca. 10-fold increase in reactivity. The results suggest that amino acid residues located close to DNA (for example, in DNA binding proteins such as histones) might participate in the repair of oxidative DNA damage. PMID- 15109271 TI - Fixed-dose combination therapy for psoriasis. AB - Fixed-dose combination therapy offers stable products containing two or more medications with different mechanisms of action and safety profiles. It is also convenient for patients since only one product rather than two or more needs to be applied. Topical corticosteroids are often the mainstay of therapy in psoriasis. Diprosalic and Nerisalic contain a topical corticosteroid (betamethasone dipropionate and diflucortolone, respectively) and salicylic acid. A left/right study showed that both products have comparable efficacy. It has also been shown that betamethasone dipropionate + salicylic acid ointment has similar efficacy to clobetasol and calcipotriene (calcipotriol) ointments. Betamethasone dipropionate + salicylic acid lotion has similar efficacy to clobetasol lotion. Faster improvement of scaling, itching, and redness was noted with betamethasone dipropionate + salicylic acid lotion compared with betamethasone dipropionate alone. Dovobet (Daivobet) ointment is a fixed-dose combination product containing betamethasone dipropionate and calcipotriene. Clinical studies have shown that it has greater efficacy and a faster speed of onset than the individual components or tacalcitol. Once daily and twice daily treatments have similar efficacy. Psoriasis Area and Severity Index reductions of approximately 40% after 1 week and 70% after 4 weeks of therapy were consistently noted in six large international studies involving >6000 patients. Betamethasone dipropionate + calcipotriene treatment is associated with approximately 75% less adverse cutaneous events as compared with tacalcitol, 50% less compared with calcipotriene, and a similar number as treatment with betamethasone dipropionate. PMID- 15109270 TI - Disulfide bond rearrangement during formation of the chorionic gonadotropin beta subunit cystine knot in vivo. AB - The intracellular kinetic folding pathway of the human chorionic gonadotropin beta-subunit (hCG-beta) reveals the presence of a disulfide between Cys residues 38-57 that is not detected by X-ray analysis of secreted hCG-beta. This led us to propose that disulfide rearrangement is an essential feature of cystine knot formation during CG-beta folding. To test this, we used disulfide bond formation to monitor progression of intracellular folding intermediates of a previously uncharacterized protein, the CG-beta subunit of cynomolgous macaque (Macaca fascicularis). Like its human counterpart hCG-beta with which it shares 81% identity, macaque (m)CG-beta is a cystine knot-containing subunit that assembles with an alpha-subunit common to all glycoprotein hormone members of its species to form a biologically active heterodimer, mCG, which, like hCG, is required for pregnancy maintenance. An early mCG-beta folding intermediate, mpbeta1, contained two disulfide bonds, one between Cys34 and Cys88 and the other between Cys38 and Cys57. The subsequent folding intermediate, mpbeta2-early, was represented by an ensemble of folding forms that, in addition to the two disulfides mentioned above, included disulfide linkages between Cys9 and Cys57 and between Cys38 and Cys90. These latter two disulfides are those contained within the beta-subunit cystine knot and reveal that a disulfide exchange occurred during the mpbeta2 early folding step leading to formation of the mCG-beta knot. Thus, while defining the intracellular kinetic protein folding pathway of a monkey homologue of CG-beta, we detected the previously predicted disulfide exchange event crucial for CG-beta cystine knot formation and attainment of CG-beta assembly competence. PMID- 15109272 TI - Topical antibacterial treatments for acne vulgaris : comparative review and guide to selection. AB - Topical antibacterial agents are an essential part of the armamentarium for treating acne vulgaris. They are indicated for mild-to-moderate acne, and are a useful alternative for patients who cannot take systemic antibacterials. Topical antibacterials such as clindamycin, erythromycin, and tetracycline are bacteriostatic for Propionibacterium acnes, and have also been demonstrated to have anti-inflammatory activities through inhibition of lipase production by P. acnes, as well as inhibition of leukocyte chemotaxis. Benzoyl peroxide is a non antibiotic antibacterial agent that is bactericidal against P. acnes and has the distinct advantage that thus far, no resistance has been detected against it. Combined agents such as erythromycin/zinc, erythromycin/tretinoin, erythromycin/isotretinoin, erythromycin/benzoyl peroxide, and clindamycin/benzoyl peroxide are increasingly being used and have been proven to be effective. They generally demonstrate good overall tolerability and are useful in reducing the development of antibacterial resistance in P. acnes. The selection of a topical antibacterial agent should be tailored for specific patients by choosing an agent that matches the patient's skin characteristics and acne type. Topical antibacterial agents should generally not be used for extended periods beyond 3 months, and topical antibacterials should ideally not be combined with systemic antibacterial therapy for acne; in particular, the use of topical and systemic antibacterials is to be avoided as far as possible. PMID- 15109273 TI - Prurigo: diagnosis and management. AB - Prurigo is a condition of nodular cutaneous lesions that itch (pruire) intensely. Although the acute form can be caused by insect stings, most of the subacute and chronic forms appear to be idiopathic. Toxic agents deposited in the skin by exogenous factors such as parasites, bacteria, or topically or orally administered drugs can induce itch. In susceptible individuals, physical mechanisms such as UV light can induce changes in epidermal innervation that result both in itch generally and in prurigo lesions. Prurigo is sometimes associated with atopy, pregnancy, internal diseases, malabsorption, or malignancy. Some forms of prurigo may be secondary to scratching. Emotional factors can also influence the perception of itch and induce prurigo by provoking scratching. These are the various specialized forms of prurigo, and there are certain others, such as prurigo pigmentosa, that have some ethnic preference. Topical treatments by corticosteroids, coal tar, bath photochemotherapy, UVB, cryotherapy, or capsaicin, as well as systemic regimens involving use of psoralen + UVA (PUVA), erythromycin, arotinoid acid, cyclosporine, chloroquine, dapsone, minocycline, naltrexone, azathioprine or thalidomide are used for the treatment of this condition. Psychotherapeutic agents to treat problems of mood that deteriorate prurigo are also prescribed. Combined sequential treatments for generalized, therapy-resistant cases need to be tailored to the exacerbations that occur and to provide maintenance treatment in order to enable the patient to withstand the intolerable itch. PMID- 15109274 TI - Levamisole in dermatology : a review. AB - Levamisole, an anthelmintic agent with a wide range of immunomodulatory actions, has been used successfully as monotherapy and an adjunct to treatment in a variety of diseases. Since 1990, combination therapy of levamisole and fluorouracil has played an important role in the treatment of resected Dukes stage C adenocarcinoma of the colon. Because of its immunomodulating effects levamisole has been used in a wide range of diseases with and without success. In dermatologic disease levamisole has been successfully used in the treatment of parasitic, viral and bacterial infections including leprosy, collagen vascular diseases, inflammatory skin diseases and children with impaired immune a variety of reasons. It has also been used in combination with other drugs for treating a number of dermatologic disorders, e.g. in combination with cimetidine for treating recalcitrant warts, with prednisolone for treating lichen planus, erythema multiforme and aphthous ulcers of the mouth. Adverse affects of levamisole are mild and infrequent and include rash, nausea, abdominal cramps, taste alteration, alopecia, arthralgia, and a flu-like syndrome. It can rarely cause agranulocytosis. More studies need to be undertaken to study the full potential of levamisole in dermatologic diseases. PMID- 15109275 TI - Vulvar lichen sclerosus : pathophysiology and treatment. AB - Lichen sclerosus is a chronic disorder of the skin and mucosal surfaces, and is most commonly seen on the female genital skin. It also occurs on other areas of the body. Any age group may be affected, although it is seen more often in elderly women. The exact cause of lichen sclerosus is unknown. There have been reports of family members with lichen sclerosus; thus it may have a genetic link. There is also the possibility of an autoimmune connection. Currently, ultra potent topical corticosteroids are the medical treatment of choice. Other treatments that have been utilized for this condition include testosterone, progesterone, tacrolimus, surgery, and phototherapy. Surgery should be reserved for symptomatic patients who fail to respond to multiple medical treatments, as there is a high recurrence rate following surgery. The risk of developing squamous cell carcinoma of the vulva approaches 5% in women with vulvar lichen sclerosus, and therefore close surveillance by the healthcare provider and patient is needed. This review discusses the history, clinical features, pathophysiology, and treatment of lichen sclerosus of the vulva, as well as pregnancy issues and sexual function in patients with this condition. In addition, problems specific to children with lichen sclerosus are reviewed. PMID- 15109276 TI - Topical methyl aminolevulinate. AB - Methyl aminolevulinate (MAL), an ester of aminolevulinic acid, has been used effectively as a topical photosensitizing agent in the photodynamic therapy (PDT) of epidermal lesions such as actinic keratosis (AK) and basal cell carcinoma (BCC). The optimal regimen for MAL-PDT (as used in all clinical trials) is MAL 160 mg/g applied for 3 hours before illumination with red light (570-670 nm) at a total light dose of 75 J/cm(2), as determined in dose-finding trials. In randomized, multicenter, phase III clinical trials, treatment with MAL-PDT resulted in a complete response (i.e. complete disappearance) in up to 91% of AK lesions and up to 97% of BCC lesions. With regard to lesion response rates, MAL PDT was superior to placebo-PDT (in AK or BCC), and at least as effective as cryotherapy (in AK or BCC) or excision surgery (in BCC). Cosmetic outcome with MAL-PDT was excellent to good in the vast majority of patients, and was judged by investigators to be better than with cryotherapy (in AK or BCC) or surgery (in BCC). black triangle Over 75% of BCC lesions treated with MAL-PDT, including difficult-to-treat BCC, were still in remission 12-24 months after the last treatment. MAL-PDT was generally well tolerated in clinical trials. The most frequently reported adverse events were local phototoxicity reactions, most of which were of mild to moderate intensity, resolved rapidly and were rarely treatment limiting. PMID- 15109281 TI - [Cerebrovascular disease risk factors and secondary prevention]. PMID- 15109282 TI - [Cerebrovascular accidents in Osona borough. Cardiovascular risks factors]. AB - INTRODUCTION: Cerebrovascular disease (CVD) is the second cause of hospitalization in the order of frequency in our service, thus reflecting this pathologys high incidence in our borough. This study analyzes clinical and biological characteristics, cardiovascular risk factors and actions taken in hospital discharge in the internal medicine department of a borough hospital (the reference hospital in Osona borough), as well as other characteristics of the hospitalized CVD patients from January 2001 to December 2001. METHOD: This study was performed by revising each patient's hospital discharge report. 277 patients were hospitalized for CVD. RESULTS: Biological and demographic characteristics, as well as cardiovascular risk factors analyzed (arterial hypertension, diabetes, smoking, or dyslipidemy) were similar to other series. Incidence of hemorrhagic and cardio-embolic CVD was slightly lower, taken into account that hemorrhagic episodes that needed neurosurgical intervention were transferred to a higher level center with a department of neurosurgery. Age was neither a factor for bad prediction leading to bad sequels after the episode nor a cause of an increased mortality. This data differs from others series. CONCLUSION: Actions taken on discharge, on cardiovascular risk factors and on hygienic-dietetic recommendations were deficient. Average stay was higher when compared to average stay in specialized ictus units. PMID- 15109283 TI - [Clinical patterns of community acquired pneumonia microbiologically documented compared to the not documented.]Prospective and comparative study between 1991 and 1997]. AB - INTRODUCTION: The diagnostic of the pneumonia is the problems more important for its adequate treatment and best evolution. OBJECTIVES: To study in a prospective way the patron clinic de la NAC compared the documented with the not documented in a microbiological way. PATIENTS Y METHODS: We have studied patients with NAC hospitalized in the HUVA between January of 1991 and May of 1997. The diagnostical criterion of pneumonia and of the hospitable ingress were the classics accepted for this infections, was doing in all cases diagnostical studies not invasive. It was analyzed the clinical patron of the NAC microbiologically documented compare to with the not documented. In the same way, the documented was divided in typical and not typical according to the microorganisms aisled. Besides was realized an statistic study using tables of contingency and test Fisher. RESULTS: It was studied 409 patients with NAC, from which 161 (39.6%) had microbiological documentation, it was found 119 micro organism typical and 42 atypical. On the one hand, the female sex, EPOC, fever, tos and purulence esputum and hyperglucaemia, were associated significantly with the documented NAC. The presence of cardiopaty, seriously initial clinical situation, gastrointestinal disorders and previous infections and use of antibiotics, were associated with not documented. The age more than 65 years, presence of comorbility, purulence sputum, pleural pain, toghether with VSG>50 and lobar infiltrate, were associated significantly to typical pneumonia, while tabaquical habit and extrapulmonary sintoms (artromialgias) were with the atypical. CONCLUSIONS: The knowledge of the clinical patrons of the NAC are great impact in the diagnostic and treatment antibiotic adecuate. PMID- 15109284 TI - [Iodine deficiency disorder persistance after introduction of iodized salt in a previously endemic goiter area]. AB - OBJECTIVE: Correction of iodine deficiency diminishes the incidence of toxic nodular goitre. The aim of this study was to assess the etiology of thyrotoxicosis in two areas with different goitre prevalence in Galicia, fifteen years after the institutional campaign of salt iodination. Results of the present survey are compared with those from a study performed ten years ago. METHODS: Two hundred and two thyrotoxic patients attended in La Coruna (coastal zone) and Lugo (inland zone) from January 2000 to May 2002 were included. Clinical and exploratory data were recorded. Thyroid hormones, thyroid-directed antibodies and thyroid stimulating immunoglobulin (TSI) were measured. A thyroid 99m technetium or 123 iodine scintigram was performed. RESULTS: In the whole group 58.6% of the cases were diagnosed of nodular goitre, 30.3% of Graves disease, 7.1% of iodine induced thyrotoxicosis and 3.5% of subacute thyroiditis. In coastal zone these percentages were 52.2, 37.0, 5.4 and 4.3%, respectively. In the inland area, 64.2% nodular goitre, 24.5% Graves disease, 8.5% iodine induced thyrotoxicosis and 2.8% subacute thyroiditis. The most frequent diagnosis in both the whole group and in each area was toxic multinodular goitre. CONCLUSIONS: In some areas of Spain, toxic nodular goitre continues to be the most frequent cause of thyrotoxicosis. It is necessary to insist on implementation and monitoring of iodine supplementation programs. PMID- 15109285 TI - HIV-associated primary body-cavity-based lymphoma: clinico-biologic features in three patients diagnosed at the same institution. AB - Primary effusion lymphoma (PEL) is a recently individualized form of non-Hodgkin lymphoma (WHO classification) that mainly develops in HIV infected males, more frequently in homosexuals and advanced stages of the disease (total CD4+ lymphocyte count below 100-200/mL). Occasionally, it appears in others immunodepressive states (such as solid organs postransplant period) and even, although very rarelly, in immunocompetents patients. From a pathogenetic point of view, PEL has been related to Kaposi's sarcoma-associated herpes virus (also named human herpesvirus 8) and to the clinical antecedent of Kaposis sarcoma. Relative unfrequency of this disease, the absence of wide casuistics allowing a better characterization, and its unfavorable outcome, support the need of a deeper knowledge. We present here the clinical-biological findings of three patients that were diagnosed of pleural PEL in our institution in the last two years. PMID- 15109286 TI - [Skeletal manifestations of Gauchers disease. A report of two cases]. AB - Gauchers disease is the most frequent hereditary lysosomal deposit storage disorder. It is characterized by a deficiency of the enzyme glucocerebrosidase that leads to an accumulation of substrate in the interior of the macrophage lysosomes. It is classified in three types, according to the presence of central nervous system involvement (type 2 and 3) or not (type 1). It is a multisystemic disease and in the majority of patients there is hepatosplenomegaly, anemia and thrombocytopenia. Skeletal involvement is also important and it is frequently the most disabling manifestation. We present two cases of Gauchers with skeletal manifestations and we review the literature. PMID- 15109287 TI - [Polytopic and recurrent reflex sympathetic dystrophy in lower limbs in two siblings]. AB - Reflex sympathetic dystrophy (RSD) has been related to a variety of inciting and predisposing factors. However, there are few reports of a familiar or genetic background in RSD. This paper describes two cases of RSD polytopic and recurrent in lower limbs of two brothers with similar HLA. PMID- 15109288 TI - [Angina-like chest pain due to gastric anisakiasis]. AB - The chest pain is one of the most common reasons for consultation of the patients seen in the emergency services of hospitals. Still being the acute coronary syndrome one of the first causes to confirmed by its important repercussions, we do not have to forget other reasons. We report a case of acute anginalike chest pain due to gastric anisakiasis. The larvae of Anisakis in the gastric mucosa were found and extracted endoscopically. Gastric anisakiasis should be included in the differential diagnosis of acute chest pain. PMID- 15109290 TI - [Resistant tuberculosis by drugs]. AB - Among all infectious diseases, tuberculosis has probably been the most frequent cause of death and morbidity in the history of humanity. On a world-wide basis, the tuberculosis (TB) represents a serious public health problem, existing great differences between developed and developing countries in terms of controlling the disease. One of the problems inherent in this disease is the resistance of Mycobacterium tuberculosis to drugs, specially the multi-resistance (defined as resistance to isoniziade and rifampicine), it represents a problem of global health. Faced with clinical suspect, its diagnosis must be immediately done and treatment must be started as soon as possible. For this purpose, we have at our disposal such effective diagnosis and therapeutic methods. However, the most important the most important action is that all these patients must be subjected to a close evolved control carried by specialized units. PMID- 15109289 TI - [Inguinal silicotic adenopathy: presentation of a case]. AB - Silicosis is a sistemic occupational disease, including in the group of pneumoconiosis, because is resulting from the inhalation of microscopic particles of crystalline silica, which deposition and histologic changes take place in lung tissue. A 30 year-old male, construction worker who presented many mediastinal, abdominal and groinal adenopathies without lung damaged. A groinal adenophaty biopsy showed the presence of numerous noncaseating granulomas containing scattered polarizable particles compatible with silica. This case has the particular feature of the exclusive extrapulmonary presentation of silicosis. PMID- 15109291 TI - [Hepatocarcinoma with leukemoid reaction]. PMID- 15109292 TI - [Occult C virus infection in cryptogenic hepatitis]. PMID- 15109293 TI - [Bone metastases as the first manifestation of hepatocarcinoma]. PMID- 15109294 TI - [Carbamacepine - induced pancreatitis]. PMID- 15109295 TI - [Human herpesvirus 6 meningoencephalitis in an immunocompetent adult patient]. PMID- 15109296 TI - [Acute respiratory failure that needs mechanical ventilation: first manifestation of myasthenia gravis]. PMID- 15109297 TI - [Pyomyositis in sternocleidomastoid muscle]. PMID- 15109298 TI - [Megaesophagus and lung fibrosis]. PMID- 15109299 TI - [Death with dignity]. PMID- 15109300 TI - [About the terminal sedation: realities and facts, not only opinions]. PMID- 15109301 TI - Octasaccharide is the minimal length unit required for efficient binding of cyclophilin B to heparin and cell surface heparan sulphate. AB - Cyclophilin B (CyPB) is a heparin-binding protein first identified as a receptor for cyclosporin A. In previous studies, we reported that CyPB triggers chemotaxis and integrin-mediated adhesion of T-lymphocytes by way of interaction with two types of binding sites. The first site corresponds to a signalling receptor; the second site has been identified as heparan sulphate (HS) and appears crucial to induce cell adhesion. Characterization of the HS-binding unit is critical to understand the requirement of HS in pro-adhesive activity of CyPB. By using a strategy based on gel mobility shift assays with fluorophore-labelled oligosaccharides, we demonstrated that the minimal heparin unit required for efficient binding of CyPB is an octasaccharide. The mutants CyPB(KKK-) [where KKK refers to the substitutions K3A(Lys3-->Ala)/K4A/K5A] and CyPB(DeltaYFD) (where Tyr14-Phe-Asp16 has been deleted) failed to interact with octasaccharides, confirming that the Y14FD16 and K3KK5 clusters are required for CyPB binding. Molecular modelling revealed that both clusters are spatially arranged so that they may act synergistically to form a binding site for the octasaccharide. We then demonstrated that heparin-derived octasaccharides and higher degree of polymerization oligosaccharides inhibited the interaction between CyPB and fluorophore-labelled HS chains purified from T-lymphocytes, and strongly reduced the HS-dependent pro-adhesive activity of CyPB. However, oligosaccharides or heparin were unable to restore adhesion of heparinase-treated T-lymphocytes, indicating that HS has to be present on the cell membrane to support the pro adhesive activity of CyPB. Altogether, these results demonstrate that the octasaccharide is likely to be the minimal length unit required for efficient binding of CyPB to cell surface HS and consequent HS-dependent cell responses. PMID- 15109302 TI - Syntaxin 5 interacts with presenilin holoproteins, but not with their N- or C terminal fragments, and affects beta-amyloid peptide production. AB - Mutations in presenilins 1 and 2 (PS1 and PS2) account for the majority of cases of early-onset familial Alzheimer's disease. However, the trafficking and interaction of PSs with other proteins in the early secretory pathways are poorly understood. Using co-immunoprecipitation, we found that PS bound to Syx5 (syntaxin 5), which is a target-soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor involved in endoplasmic reticulum (ER)-Golgi vesicular transport in vivo. Syx5 interacted only with the full-length PS holoproteins and not with the naturally occurring N- or C-terminal fragments. The PS holoproteins co-immunoprecipitated with the mutant Syx5, which localized to the ER and Golgi compartments, despite the substitution of the transmembrane region with that of syntaxin 1A. In contrast, the transmembrane deletion mutant that localized to the cytosol, but not to the ER or Golgi compartments, did not co-immunoprecipitate the PS holoproteins. The PS1 variant linked to familial Alzheimer's disease (PS1DeltaE9), lacking the region that contains the endoproteolytic cleavage site in the cytoplasmic loop, showed markedly decreased binding to Syx5. Immunofluorescence and sucrose-density-gradient fractionation analyses showed that the full-length PS holoproteins co-localized with Syx5 to the ER and cis-Golgi compartments. Furthermore, Syx5 overexpression resulted in the accumulation of PS holoproteins and the beta-amyloid precursor protein, and reduced the secretion of the Abeta (amyloid beta) peptide in COS-7 cells. In summary, these results indicate that Syx5 binds to full-length PSs and affects the processing and trafficking of beta-amyloid precursor protein in the early secretory compartments. PMID- 15109303 TI - Alternative reading frame protein (ARF)-independent function of CARF (collaborator of ARF) involves its interactions with p53: evidence for a novel p53-activation pathway and its negative feedback control. AB - CARF, a collaborator of ARF (alternative reading frame protein), was cloned as a novel ARF-binding protein from a yeast-interaction screen. It potentiated ARF mediated p53 function, and also caused a moderate increase in p53 activity in the absence of ARF. We herein report the molecular mechanism of ARF-independent function of CARF. By employing a variety of approaches, including overexpression of CARF, its suppression by small interfering RNA and use of protease inhibitors, we demonstrate that: (i) CARF directly interacts with wild-type p53, causing its stabilization and functional activation; and (ii) CARF and p53 levels show an inverse relationship that is instigated by a negative-feedback control via a proteasome-mediated degradation pathway. PMID- 15109304 TI - Reactive oxygen species play no role in the candidacidal activity of the salivary antimicrobial peptide histatin 5. AB - The mechanism of action of antimicrobial peptides is still a matter of debate. The formation of ROS (reactive oxygen species) has been suggested to be the crucial step in the fungicidal mechanism of a number of antimicrobial peptides, including histatin 5 and lactoferrin-derived peptides. In the present study we have investigated the effects of histatin 5 and of a more amphipathic synthetic derivative, dhvar4, on the generation of ROS in the yeast Candida albicans, using dihydroethidium as an indicator for ROS. With both peptides, a substantial enhancement of fluorescence was observed. However, TEMPO (2,2,6,6 tetramethylpiperidine-N-oxyl), a cell-permeant ROS scavenger, did not have an inhibitory effect on killing or on the enhancement of fluorescence. Furthermore, antimycin and azide, which have been reported to induce ROS in vitro, were not able to enhance the dihydroethidium fluorescence, while chlorhexidine, a non specific antiseptic agent, enhanced dihydroethidium fluorescence to the same extent as did the peptides. Fluorescence microscopy showed the fluorescence enhancement to be a consequence of the release of unbound preformed ethidium from the mitochondrial matrix within the cell. It is concluded that ROS do not play a role in the histatin 5-mediated killing of C. albicans. PMID- 15109305 TI - Regulation of the MST1 kinase by autophosphorylation, by the growth inhibitory proteins, RASSF1 and NORE1, and by Ras. AB - MST1 (mammalian Sterile20-like 1) and MST2 are closely related Class II GC (protein Ser/Thr) kinases that initiate apoptosis when transiently overexpressed in mammalian cells. In the present study, we show that recombinant MST1/2 undergo a robust autoactivation in vitro, mediated by an intramolecular autophosphorylation of a single site [MST1(Thr183)/MST2(Thr180)] on the activation loop of an MST dimer. Endogenous full-length MST1 is activated by a variety of stressful stimuli, accompanied by the secondary appearance of a 36 kDa Thr183-phosphorylated, caspase-cleaved catalytic fragment. Recombinant MST1 exhibits only 2-5% activation during transient expression; endogenous MST1 in the cycling HeLa or KB cells has a similar low fractional activation, but 2 h incubation with okadaic acid (1 mM) results in 100% activation. Endogenous MST1 immunoprecipitated from KB cells is specifically associated with substoichiometric amounts of the growth inhibitory polypeptides RASSF1A and NORE1A (novel Ras effector 1A; a Ras-GTP-binding protein). Co-expression of RASSF1A, RASSF1C, NORE1A and NORE1B with MST1 markedly suppresses MST1(Thr183) phosphorylation in vivo and abolishes the ability of MST1 to undergo Mg-ATP mediated autoactivation in vitro; direct addition of purified NORE1A in vitro also inhibits MST1 activation. In contrast, co-transfection of MST1 with NORE1A modified by the addition of a C-terminal CAAX motif results in a substantial increase in MST1(Thr183) phosphorylation, as does fusion of a myristoylation motif directly on to the MST1 N-terminus. Moreover, MST1 polypeptides, bound via wild-type NORE1A to Ras(G12V) (where G12V stands for Gly12Val), exhibit higher Thr183 phosphorylation compared with MST1 bound to NORE1A alone. Nevertheless, serum stimulation of KB cells does not detectably increase the activation state of endogenous MST1 or MST2 despite promoting the recruitment of the endogenous NORE1-MST1 complex to endogenous Ras. We propose that the NORE1/RASSF1 polypeptides, in addition to their role in maintaining the low activity of MST1 in vivo, direct MST1 to sites of activation and perhaps co-localization with endogenous substrates. PMID- 15109306 TI - Control of expression of the lectin-like protein Reg-1 by gastrin: role of the Rho family GTPase RhoA and a C-rich promoter element. AB - The expression of members of the Reg family of secreted lectin-like proteins is increased in response to stress, inflammation and damage in many tissues. In the stomach, Reg is located in enterochromaffin-like cells, where its expression is stimulated by the gastric hormone gastrin. We have examined the mechanisms by which gastrin stimulates expression of Reg-1. Deletional mutations of 2.1 to 0.1 kb of the rat Reg-1 promoter in a luciferase reporter vector were transiently transfected into gastric cancer AGS-G(R) cells. All promoter fragments tested showed similar relative increases in luciferase expression in response to gastrin (1 nM). The response to gastrin of the smallest (104 bp) construct was 4.2+/-0.4 fold over basal. These responses were reduced by Ro-32-0432, a protein kinase C inhibitor, by C3-transferase, a Clostridium botulinum toxin and a selective inhibitor of the Rho family GTPase RhoA, and by co-transfection with a dominant negative form of RhoA. Co-transfection with a constitutively active form of RhoA stimulated expression 11.6+/-1.7-fold over basal. Mutations through the 104 bp construct identified a C-rich element (C-79CCCTCCC-72) required for responses to gastrin, PKC (protein kinase C) and L63RhoA (the constitutively active form of human RhoA protein containing a glutamine-to-leucine substitution at position 63). EMSAs (electrophoretic-mobility-shift assays) using nuclear extracts of control and gastrin-stimulated AGS-G(R) cells and a probe spanning -86 to -64 bp revealed multiple binding proteins. There was no effect of gastrin on the pattern of binding. Supershift assays indicated that transcription factors Sp1 and Sp3 bound the C-rich sequence. We conclude that gastrin stimulates Reg expression via activation of PKC and RhoA, that a C-rich region (-79 to -72) is critical for the response and that Sp-family transcription factors bind to this region of the promoter. PMID- 15109307 TI - Analysis of the ERK1,2 transcriptome in mammary epithelial cells. AB - MAPK (mitogen-activated protein kinase) pathways constitute major regulators of cellular transcriptional programmes. We analysed the ERK1,2 (extracellular-signal regulated kinase 1,2) transcriptome in a non-transformed MEC (mammary epithelial cell) line, MCF-12A, utilizing rAd MEK1EE, a recombinant adenovirus encoding constitutively active MEK1 (MAPK/ERK kinase 1). rAd MEK1EE infection induced morphological changes and DNA synthesis which were inhibited by the MEK1,2 inhibitor PD184352. Hierarchical clustering of data derived from seven time points over 24 h identified 430 and 305 co-ordinately up-regulated and down regulated genes respectively. c-Myc binding sites were identified in the promoters of most of these up-regulated genes. A total of 46 candidate effectors of the Raf/MEK/ERK1,2 pathway in MECs were identified by comparing our dataset with previously reported Raf-1-regulated genes. These analyses led to the identification of a suite of growth factors co-ordinately induced by MEK1EE, including multiple ErbB ligands, vascular endothelial growth factor and PHRP (parathyroid hormone-related protein). PHRP is the primary mediator of humoral hypercalcaemia of malignancy, and has been implicated in metastasis to bone. We demonstrate that PHRP is secreted by MEK1EE-expressing cells. This secretion is inhibited by PD184352, but not by ErbB inhibitors. Our results suggest that, in addition to anti-proliferative properties, MEK1,2 inhibitors may be anti angiogenic and possess therapeutic utility in the treatment of PHRP-positive tumours. PMID- 15109308 TI - Sphingosine kinase activity is required for sphingosine-mediated phospholipase D activation in C2C12 myoblasts. AB - Sphingosine (Sph) has been implicated as a modulator of membrane signal transduction systems and as a regulatory element of cardiac and skeletal muscle physiology, but little information is presently available on its precise mechanism of action. Recent studies have shown that sphingosine 1-phosphate (S1P), generated by the action of sphingosine kinase (SphK) on Sph, also possesses biological activity, acting as an intracellular messenger, as well as an extracellular ligand for specific membrane receptors. At present, however, it is not clear whether the biological effects elicited by Sph are attributable to its conversion into S1P. In the present study, we show that Sph significantly stimulated phospholipase D (PLD) activity in mouse C2C12 myoblasts via a previously unrecognized mechanism that requires the conversion of Sph into S1P and its subsequent action as extracellular ligand. Indeed, Sph-induced activation of PLD was inhibited by N,N-dimethyl-D-erythro-sphingosine (DMS), at concentrations capable of specifically inhibiting SphK. Moreover, the crucial role of SphK-derived S1P in the activation of PLD by Sph was confirmed by the observed potentiated effect of Sph in myoblasts where SphK1 was overexpressed, and the attenuated response in cells transfected with the dominant negative form of SphK1. Notably, the measurement of S1P formation in vivo by employing labelled ATP revealed that cell-associated SphK activity in the extracellular compartment largely contributed to the transformation of Sph into S1P, with the amount of SphK released into the medium being negligible. It will be important to establish whether the mechanism of action identified in the present study is implicated in the multiple biological effects elicited by Sph in muscle cells. PMID- 15109309 TI - Effect of a biopsychosocial approach on patient satisfaction and patterns of care. AB - BACKGROUND: There is a growing tendency to include in medical curricula teaching programs that promote a biopsychosocial (BPS) approach to patient care. However, we know of no attempts to assess their effect on patterns of care and health care expenditures. OBJECTIVE: To determine whether 1) a teaching intervention aiming to promote a BPS approach to care affects the duration of the doctor-patient encounter, health expenditures, and patient satisfaction with care, and 2) the teaching method employed affects these outcomes. METHODS: We compared two teaching methods. The first one (didactic) consisted of reading assignments, lectures, and group discussions. The second (interactive) consisted of reading assignments, small group discussions, Balint groups, and role-playing exercises. We videotaped patient encounters 1 month before and 6 months after the teaching interventions, and recorded the duration of the videotaped encounters and whether the doctor had prescribed medications, ordered tests, and referred the patient to consultants. Patient satisfaction was measured by a structured questionnaire. RESULTS: Both teaching interventions were followed by a reduction in medications prescribed and by improved patient satisfaction. Compared to the didactic group, the interactive group prescribed even fewer medications, ordered fewer laboratory examinations, and elicited higher scores of patient satisfaction. The average duration of the encounters after the didactic and interactive teaching interventions was longer than that before by 36 and 42 seconds, respectively. CONCLUSIONS: A BPS teaching intervention may reduce health care expenditures and enhance patients' satisfaction, without changing markedly the duration of the encounter. An interactive method of instruction was more effective in achieving these objectives than a didactic one. PMID- 15109310 TI - Meeting requirements and changing culture. The development of a web-based clinical skills evaluation system. AB - The Accreditation Council of Graduate Medical Education (ACGME) and the Residency Review Committee require a competency-based, accessible evaluation system. The paper system at our institution did not meet these demands and suffered from low compliance. A diverse committee of internal medicine faculty, program directors, and house staff designed a new clinical evaluation strategy based on ACGME competencies and utilizing a modular web-based system called ResEval. ResEval more effectively met requirements and provided useful data for program and curriculum development. The system is paperless, allows for evaluations at any time, and produces customized evaluation reports, dramatically improving our ability to analyze evaluation data. The use of this novel system and the inclusion of a robust technology infrastructure, repeated training and e-mail reminders, and program leadership commitment resulted in an increase in clinical skills evaluations performed and a rapid change in the workflow and culture of evaluation at our residency program. PMID- 15109311 TI - Teaching and assessing resident competence in practice-based learning and improvement. AB - We designed, implemented, and evaluated a 4-week practice-based learning and improvement (PBLI) elective. Eleven internal medicine residents from 2 separate residency programs participated in the PBLI elective and 22 other residents comprised a comparison group. Residents in each group had similar pretest Quality Improvement Knowledge Application Tool scores; but after the PBLI elective, participant scores were significantly higher. Also, participants' self-assessed ratings of PBLI skills increased after the rotation and remained elevated 6 months afterward. In this curriculum, residents completed a project to improve patient care and demonstrated their knowledge on an evaluation tool in a way that was superior to nonparticipants. PMID- 15109312 TI - Toward an informal curriculum that teaches professionalism. Transforming the social environment of a medical school. AB - The social environment or "informal" curriculum of a medical school profoundly influences students' values and professional identities. The Indiana University School of Medicine is seeking to foster a social environment that consistently embodies and reinforces the values of its formal competency-based curriculum. Using an appreciative narrative-based approach, we have been encouraging students, residents, and faculty to be more mindful of relationship dynamics throughout the school. As participants discover how much relational capacity already exists and how widespread is the desire for a more collaborative environment, their perceptions of the school seem to shift, evoking behavior change and hopeful expectations for the future. PMID- 15109313 TI - Effect of an Internet-based curriculum on postgraduate education. A multicenter intervention. AB - We hypothesized that the Internet could be used to disseminate and evaluate a curriculum in ambulatory care, and that internal medicine residency program directors would value features made possible by online dissemination. An Internet based ambulatory care curriculum was developed and marketed to internal medicine residency program directors. Utilization and knowledge outcomes were tracked by the website; opinions of program directors were measured by paper surveys. Twenty four programs enrolled with the online curriculum. The curriculum was rated favorably by all programs, test scores on curricular content improved significantly, and program directors rated highly features made possible by an Internet-based curriculum. PMID- 15109314 TI - Creation of an innovative inpatient medical procedure service and a method to evaluate house staff competency. AB - INTRODUCTION: Training residents in medical procedures is an area of growing interest. Studies demonstrate that internal medicine residents are inadequately trained to perform common medical procedures, and program directors report residents do not master these essential skills. The American Board of Internal Medicine requires substantiation of competence in procedure skills for all internal medicine residents; however, for most procedures, standards of competence do not exist. OBJECTIVE: 1) Create a new and standardized approach to teaching, performing, and evaluating inpatient medical procedures; 2) Determine the number of procedures required until trainees develop competence, by assessing both clinical knowledge and psychomotor skills; 3) Improve patient safety. DESIGN: A Medical Procedure Service (MPS), consisting of select faculty who are experts at common inpatient procedures, was established to supervise residents performing medical procedures. Faculty monitor residents' psychomotor performance, while clinical knowledge is taught through a complementary, comprehensive curriculum. After the completion of each procedure, the trainee and supervising faculty member independently complete online questionnaires. RESULTS: During this pilot program, 246 procedures were supervised, with a pooled major complication rate of 3.7%. 123 thoracenteses were supervised, with a pneumothorax rate of 3.3%; this compares favorably with a pooled analysis of the literature. 87% of surveyed house staff felt the procedure service helped in their education of medical procedures. CONCLUSIONS: The "see one, do one, teach one" model of procedure education is dangerously inadequate. Through the development of a Medical Procedure Service, and an associated procedure curriculum and a mechanism of evaluation, we hope to reduce the rate of complications and errors related to medical procedures and to determine at what point competency is achieved for these procedures. PMID- 15109315 TI - Cultural consensus analysis as a tool for clinic improvements. AB - Some problems in clinic function recur because of unexpected value differences between patients, faculty, and residents. Cultural consensus analysis (CCA) is a method used by anthropologists to identify groups with shared values. After conducting an ethnographic study and using focus groups, we developed and validated a CCA tool for use in clinics. Using this instrument, we identified distinct groups with 6 important value differences between those groups. An analysis of these value differences suggested specific and pragmatic interventions to improve clinic functioning. The instrument has also performed well in preliminary tests at another clinic. PMID- 15109316 TI - Teaching and learning in an 80-hour work week: a novel day-float rotation for medical residents. AB - The 80-hour workweek limit for residents provides an opportunity for residency directors to creatively innovate their programs. Our novel day-float rotation augmented both the educational structure within the inpatient team setting and the ability for house staff to complete their work within the mandated limits. Descriptive evaluation of the rotation was performed through an end-of-rotation questionnaire. The average length of the ward residents' work week was quantified before and after the rotation's implementation. Educational portfolios and mentored peer-teaching opportunities enriched the rotation. As measured by our evaluation, this new rotation enhanced learning and patient care while reducing work hours for inpatient ward residents. PMID- 15109317 TI - A primary care musculoskeletal clinic for residents: success and sustainability. AB - Musculoskeletal complaints are common, but are often underemphasized in residency training. We evaluated the experience of residents (12) in 4 sessions of an innovative concentrated ambulatory, community-based musculoskeletal (MS) clinic precepted by general internists with additional training in teaching MS medicine. Compared with the year long longitudinal house staff (HS) clinic experience, the mean number of musculoskeletal diagnoses per resident seen in MS clinic was higher (13.9 [standard deviation 4.0] vs 5.4 [standard deviation 4.0]; P <.01). Common diagnoses in MS clinic included shoulder, hip, and knee tendonitis/bursitis, and the majority of diagnoses in HS clinic were nonspecific arthralgia (66%). Fifty-two injections were performed in MS clinic over the year, compared with one in HS clinic. PMID- 15109318 TI - Measuring the competence of residents as teachers. AB - Medical residents, frontline clinical educators, must be competent teachers. Typically, resident teaching competence is not assessed through any other means than gleaning learner's comments. We developed, evaluated, and integrated into our annual objective structured clinical examination a resident teaching skills assessment using "standardized" students. Faculty observers rated residents using a customized 19-item rating instrument developed to assess teaching competencies that were identified and defined as part of our project. This was feasible, acceptable, and valuable to all 65 residents, 8 students, and 16 faculty who participated. Teaching scenarios have potential as reliable, valid, and practical measures of resident teaching skills. PMID- 15109319 TI - Teaching patient-centered tobacco intervention to first-year medical students. AB - The University of Wisconsin's Tobacco Intervention Basic Skills curriculum (TIBS) was inaugurated to begin training 147 first-year medical students in skills for promoting health behavior change. Learning activities included lecture, demonstration, reading, quiz, role-play exercises, and standardized patient interviews. After TIBS, the 69 students who provided pre- and postintervention data exhibited more therapeutic attitudes and increased knowledge and self confidence in applying TIBS skills. Two months later, 52% of the 109 posttest respondents had applied TIBS in clinical settings, often for behaviors other than tobacco use. We conclude that medical students can gain from early training on promoting behavior change. PMID- 15109320 TI - A workshop to teach medical students communication skills and clinical knowledge about end-of-life care. AB - We describe a half-day workshop to teach third-year medical students three focused end-of-life care skills: breaking bad news, discussing advance directives, and assessing and managing pain. Our workshop included a readers' theater exercise and three role-play exercises. In two of the workshops, faculty members played the role of patients. We used readers' theater to engage the students on an emotional level and set a reflective tone for the workshop. Evaluations reflected that most respondents felt that the workshop enhanced their understanding and ability to address these skills with patients. By 6 months, many students reported applying these skills to patient care in a way they thought was effective. PMID- 15109321 TI - Competency-based learning: the impact of targeted resident education and feedback on Pap smear adequacy rates. AB - Little is known about assessing or improving competency in Papanicolau (Pap) smear sampling among internal medicine residents. We hypothesized that a 3-part targeted resident physician educational program (educational presentation by a knowledgeable instructor, skills workshop, and peer comparison feedback) would be effective in increasing the quality of Pap smears obtained by internal medicine residents. We conducted a randomized, pre-post comparison study over a 16-month period to assess the effect of our educational intervention. We found no difference in baseline adequacy rates. Residents who received the intervention were twice as likely to obtain an adequate Pap smear. Our results suggest that a brief multifaceted intervention designed to improve the frequency with which internal medicine residents obtain endocervical cells while performing Pap smears is effective. PMID- 15109322 TI - Improving students' sexual history inquiry and HIV counseling with an interactive workshop using standardized patients. AB - Sexual history and HIV counseling are essential clinical skills. Our project's purpose was to evaluate a standardized patient (SP) educational intervention teaching third-year medical students sexual history taking and HIV counseling. A 4-hour SP workshop was delivered to one-half of the class. Four weeks later, all students engaged in an SP examination including one station on assessing sexual history taking and HIV counseling. Workshop participants scored one standard deviation higher on sexual history and HIV counseling items than nonparticipants. Our sexual history and HIV counseling curriculum was associated with students asking more thorough sexual histories and providing more HIV counseling. PMID- 15109323 TI - Clinician-teachers' self-assessments versus learners' perceptions. AB - Understanding how clinician-teachers' self-assessments compare to learners' impressions can serve to help educators place each of these evaluations in the appropriate context. Past participants of the Johns Hopkins Faculty Development Program and other physician-teachers were surveyed in 2002 regarding their teaching skills and behaviors. We surveyed their learners to compare teacher and learner assessments of teaching proficiency, behaviors, enjoyment, and career satisfaction. In each area, learners' ratings were statistically significantly higher than their teachers' self-ratings. Though it is unclear whether teachers' or learners' assessments are a more accurate reflection of the truth, the more positive learner ratings should promote self-confidence in clinician-educators regarding their teaching abilities. PMID- 15109324 TI - Feedback and the mini clinical evaluation exercise. AB - We studied the nature of feedback given after a miniCEX. We investigated whether the feedback was interactive; specifically, did the faculty allow the trainee to react to the feedback, enable self-assessment, and help trainees to develop an action plan for improvement. Finally, we investigated the number of types of recommendations given by faculty. One hundred and seven miniCEX feedback sessions were audiotaped. The faculty provided at least 1 recommendation for improvement in 80% of the feedback sessions. The majority of the sessions (61%) involved learner reaction, but in only 34% of the sessions did faculty ask for self assessment from the intern and only 8% involved an action plan from the faculty member. Faculty are using the miniCEX to provide recommendations and often encourage learner reaction, but are underutilizing other interactive feedback methods of self-assessment and action plans. Programs should consider both specific training in feedback and changes to the miniCEX form to facilitate interactive feedback. PMID- 15109325 TI - Are continuity clinic patients less satisfied when the resident is postcall? AB - Due to recent public debate and newly imposed resident work hour restrictions, we decided to investigate the relationship of resident call status to their ambulatory patients' satisfaction. Resident continuity clinic patients were asked to rate their level of satisfaction on a 10-point Likert-type scale. Using multiple regression approaches, these data were then assessed as a function of resident call status. We found that in 646 patient encounters, patient satisfaction scores were significantly less when the resident was postcall, 8.99 +/- 1.8, than when not postcall, 9.31 +/- 1.3. We herein discuss etiologies and implications of these findings for both patient care and medical education. PMID- 15109326 TI - Changing attitudes toward homeless people. AB - We assessed the impact of a 2-week required rotation in homeless health care on primary care residents' attitudes toward homeless people. Attitudes were assessed before and after the course using the Attitudes Toward Homelessness Inventory (ATHI), an instrument previously validated among undergraduate students. Attitude scores on the ATHI improved from 46 to 52 (range of possible scores 11 to 66; P =.001). The ATHI subscales showed, after the course, that residents had a greater belief that homelessness had societal causes and felt more comfortable affiliating with homeless people. After the course, residents also reported an increased interest in volunteering with homeless populations on an anonymous survey. PMID- 15109327 TI - A year of mentoring in academic medicine: case report and qualitative analysis of fifteen hours of meetings between a junior and senior faculty member. AB - We describe a specific mentoring approach in an academic general internal medicine setting by audiotaping and transcribing all mentoring sessions in the year. In advance, the mentor recorded his model. During the year, the mentee kept a process journal. Qualitative analysis revealed development of an intimate relationship based on empathy, trust, and honesty. The mentor's model was explicitly intended to develop independence, initiative, improved thinking, skills, and self-reflection. The mentor's methods included extensive and varied use of questioning, active listening, standard setting, and frequent feedback. During the mentoring, the mentee evolved as a teacher, enhanced the creativity in his teaching, and matured as a person. Specific accomplishments included a national workshop on professional writing, an innovative approach to inpatient attending, a new teaching skills curriculum for a residency program, and this study. A mentoring model stressing safety, intimacy, honesty, setting of high standards, praxis, and detailed planning and feedback was associated with mentee excitement, personal and professional growth and development, concrete accomplishments, and a commitment to teaching. PMID- 15109328 TI - A literature review of "resident-as-teacher" curricula: do teaching courses make a difference? AB - OBJECTIVES: To examine the evaluation methods of resident teaching courses and to estimate the effectiveness of these teaching courses. DESIGN: We searched the literature from 1975 to May 2003 using the PubMed MESH terms internship and residency and teaching; 1,436 articles were identified and 77 contained information regarding teaching courses. Fourteen articles contained information regarding outcomes of resident teaching courses and were selected for intensive review. MAIN RESULTS: Five uncontrolled pre-post studies used resident self reported teaching skills/behaviors as outcome measures; all reported some improvement in self-reported skills. Three uncontrolled pre-post studies examined live or videotaped resident teaching encounters and all revealed improvement in some teaching skills. One uncontrolled trial and three nonrandomized controlled trials used learner evaluations of resident teaching behaviors as outcomes and all revealed an improvement in ratings of residents after course participation. Four randomized controlled trials of resident teaching curricula are included in this review. One study did not show any quantitative benefit of a resident teaching course on performance on an objective structured teaching evaluation. Two studies assessing resident teaching evaluations before and after course participation showed conflicting results. One study noted improvements in resident teaching skills assessed through videotape analysis. CONCLUSIONS: Resident teaching courses improve resident self-assessed teaching behaviors and teaching confidence. Teaching courses are linked to improved student evaluations. Further studies must be completed to elucidate the best format, length, timing, and content of resident teaching courses and to determine whether they have an effect on learner performance. PMID- 15109329 TI - Diving for PERLS: working and performance portfolios for evaluation and reflection on learning. AB - Professional competence requires a commitment to lifelong learning, self assessment, and excellence. Complex skills such as these require flexible and comprehensive teaching and assessment measures. We describe a combination of working and performance portfolios that both foster and evaluate the development of professional competence. We explain the conceptual and practical underpinnings that maximize the effectiveness of these tools. Drawing on experience with University of Washington residents, we identify 5 criteria that can help promote successful use of portfolios: separate working and performance functions of portfolios, developing a supportive climate, developing skills in faculty and residents, observing progress over time, and fostering mentorship opportunities. PMID- 15109330 TI - Writing more informative letters of reference. AB - Writing a meaningful and valuable letter of reference is not an easy task. Several factors influence the quality of any letter of reference. First, the accuracy and reliability of the writer's impressions and judgment depend on how well he knows the individual being described. Second, the writer's frame of reference, which is determined by the number of persons at the same level that he has worked with, will impact the context and significance of his beliefs and estimations. Third, the letter-writing skills of the person composing the letter will naturally affect the letter. To support the other components of a candidate's application, a letter of reference should provide specific examples of how an individual's behavior or attitude compares to a reference group and should assess "intangibles" that are hard to glean from a curriculum vitae or from test scores. This report offers suggestions that should help physicians write more informative letters of reference. PMID- 15109331 TI - Developing a peer review process for web-based curricula: minting a new coin of the realm. AB - The World Wide Web creates new challenges and opportunities for medical educators. Prominent among these are the lack of consistent standards by which to evaluate web-based educational tools. We present the instrument that was used to review web-based innovations in medical education submissions to the 2003 Society of General Internal Medicine (SGIM) national meeting, and discuss the process used by the SGIM web-based clinical curriculum interest group to develop the instrument. The 5 highest-ranked submissions are summarized with commentary from the reviewers. PMID- 15109332 TI - Internet resources for curriculum development in medical education: an annotated bibliography. AB - Curriculum development in medical education should be a methodical and scholarly, yet practical process that addresses the needs of trainees, patients, and society. To be maximally efficient and effective, it should build upon previous work and use existing resources. A conventional search of the literature is necessary, but insufficient for this purpose. The internet provides a rich source of information and materials. This bibliography is a guide to internet resources that are of use to curriculum developers, organized into 1) medical accreditation bodies, 2) topic-oriented resources, 3) general educational resources within medicine, and 4) general education resources beyond medicine. PMID- 15109333 TI - An emerging renaissance in medical education. PMID- 15109334 TI - The "new revolution" in medical education: fostering professionalism and patient centered communication in the contemporary environment. PMID- 15109335 TI - "May we live in interesting times"--Society of General Internal Medicine clinician-educators respond to new challenges in graduate medical education. PMID- 15109336 TI - What effect does inpatient physician specialty and experience have on clinical outcomes and resource utilization on a general medical service? AB - OBJECTIVE: To examine the effects of internal medicine specialty and physician experience on inpatient resource use and clinical outcomes on an academic general medicine service. DESIGN: A 1-year retrospective cohort study. SETTING: The University of Michigan Hospitals, Ann Arbor, Michigan. PATIENTS: Two thousand six hundred seventeen admissions to the general medicine service from July 2001 to June 2002, excluding those for whom data were incomplete (n = 18). MEASUREMENTS AND MAIN RESULTS: Length of stay (LOS) and total hospital costs were used to measure resource utilization. Hospital mortality and 14-day and 30-day readmission rates were used to measure clinical outcomes. Adjusted mean LOS was significantly greater for rheumatologists (0.56 days greater; P =.002) and endocrinologists (0.38 days greater; P =.03) compared to general internists. Total costs were lower for general internists compared to endocrinologists ($1100 lower; P =.01) and rheumatologists ($431 lower; P =.07). Hospitalists showed a trend toward reduced LOS compared to all other physicians (0.31 days lower; P =.06). The top two deciles of physicians stratified by recent inpatient general medical experience showed significantly reduced LOS compared to all other physicians (0.35 days lower; P =.04). No significant differences were seen in readmission rates or in-hospital mortality among the various physician groups. CONCLUSIONS: General internists had lower lengths of stay and costs compared to endocrinologists and rheumatologists. Hospitalists showed a trend toward reduced LOS compared to all other physicians. Recent inpatient general medicine experience appears to be a determinant of reduced inpatient resource use. PMID- 15109337 TI - The impact of evidence on physicians' inpatient treatment decisions. AB - OBJECTIVE: Previous studies have shown that most medical inpatients receive treatment supported by strong evidence (evidence-based treatment), but they have not assessed whether and how physicians actually use evidence when making their treatment decisions. We investigated whether physicians would change inpatient treatment if presented with the results of a literature search. DESIGN: Before after study. SETTING: Large public teaching hospital. PARTICIPANTS: Random sample of 146 inpatients cared for by 33 internal medicine attending physicians. INTERVENTIONS: After physicians committed to a specific diagnosis and treatment plan, investigators performed standardized literature searches and provided the search results to the attending physicians. MEASUREMENTS AND MAIN RESULTS: The primary study outcome was the number of patients whose attending physicians would change treatment due to the literature searches. These changes were evaluated by blinded peer review. A secondary outcome was the proportion of patients who received evidence-based treatment before and after the literature searches. Attending physicians changed treatment for 23 (18%) of 130 eligible patients (95% confidence interval, 12% to 24%) as a result of the literature searches. Overall, 86% of patients (112 of 130) received evidence-based treatments before the searches and 87% (113 of 130) after the searches. Changes were not related to whether patients were receiving evidence-based treatment before the search (P =.6). Panels of peer reviewers judged the quality of patient care as improved or maintained for 18 (78%) of the 23 patients with treatment changes. CONCLUSIONS: Searching the literature could improve the treatment of many medical inpatients, including those already receiving evidence-based treatment. PMID- 15109338 TI - Practice, clinical management, and financial arrangements of practicing generalists. AB - OBJECTIVE: To describe the practice settings, financial arrangements, and management strategies experienced by generalist physicians and identify factors associated with reporting pressure to limit referrals, pressure to see more patients, and career dissatisfaction. DESIGN: Cross-sectional mail survey. PARTICIPANTS AND SETTING: Six hundred nineteen generalist physicians (62% response rate) caring for managed care patients in 3 Minnesota health plans during 1999. MEASUREMENTS AND MAIN RESULTS: Twenty-six percent of physicians reported pressure to limit referrals. In adjusted analyses, female physicians and those who were board certified acted as gatekeepers for most of their patients, received incentives based on performance reports and quality profiles, and received direct income from capitation, and were more likely than others to report this pressure (all P <.05). Sixty-two percent reported pressure to see more patients. In adjusted analyses, this pressure was more frequent among physicians in practices owned by health systems, those using physician extenders, and among physicians paid by salary with performance adjustment or those receiving at least some capitation (all P <.05). One-quarter (24%) of physicians were dissatisfied with their career in medicine. In adjusted analyses, physicians reporting pressure to limit referrals (risk ratio, 1.12; 95% confidence interval, 1.01 to 1.19) and those reporting pressure to see more patients (risk ratio, 1.37; 95% confidence interval, 1.08 to 1.66) were more likely to be dissatisfied than other physicians. CONCLUSIONS: Pressures to limit referrals and to see more patients are common, particularly among physicians paid based on productivity or capitation, and they are associated with career dissatisfaction. Whether future changes in practice arrangements or compensation strategies can decrease such physician-reported pressures, and ultimately improve physician satisfaction, will be an important area for future study. PMID- 15109339 TI - Tobacco industry research on smoking cessation. Recapturing young adults and other recent quitters. AB - BACKGROUND: Smoking rates are declining in the United States, except for young adults (age 18 to 24). Few organized programs target smoking cessation specifically for young adults, except programs for pregnant women. In contrast, the tobacco industry has invested much time and money studying young adult smoking patterns. Some of these data are now available in documents released through litigation. OBJECTIVE: Review tobacco industry marketing research on smoking cessation to guide new interventions and improve clinical practice, particularly to address young adult smokers' needs. METHODS: Analysis of previously secret tobacco industry documents. RESULTS: Compared to their share of the smoking population, young adult smokers have the highest spontaneous quitting rates. About 10% to 30% of smokers want to quit; light smokers and brand switchers are more likely to try. Tobacco companies attempted to deter quitting by developing products that appeared to be less addictive or more socially acceptable. Contrary to consumer expectations, "ultra low tar" cigarette smokers were actually less likely to quit. CONCLUSIONS: Tobacco industry views of young adult quitting behavior contrast with clinical practice. Tobacco marketers concentrate on recapturing young quitters, while organized smoking cessation programs are primarily used by older smokers. As young people have both the greatest propensity to quit and the greatest potential benefits from smoking cessation, targeted programs for young adults are needed. Tobacco marketing data suggest that aspirational messages that decrease the social acceptability of smoking and support smoke-free environments resonate best with young adult smokers' motivations. PMID- 15109340 TI - A traditionally administered short course failed to improve medical students' diagnostic performance. A quantitative evaluation of diagnostic thinking. AB - BACKGROUND: Quite often medical students or novice residents have difficulty in ruling out diseases even though they are quite unlikely and, due to this difficulty, such students and novice residents unnecessarily repeat laboratory or imaging tests. OBJECTIVE: To explore whether or not a carefully designed short training course teaching Bayesian probabilistic thinking improves the diagnostic ability of medical students. PARTICIPANTS AND METHODS: Ninety students at 2 medical schools were presented with clinical scenarios of coronary artery disease corresponding to high, low, and intermediate pretest probabilities. The students' estimates of test characteristics of exercise stress test, and pretest and posttest probability for each scenario were evaluated before and after the short course. RESULTS: The pretest probability estimates by the students, as well as their proficiency in applying Bayes's theorem, were improved in the high pretest probability scenario after the short course. However, estimates of pretest probability in the low pretest probability scenario, and their proficiency in applying Bayes's theorem in the intermediate and low pretest probability scenarios, showed essentially no improvement. CONCLUSION: A carefully designed, but traditionally administered, short course could not improve the students' abilities in estimating pretest probability in a low pretest probability setting, and subsequently students remained incompetent in ruling out disease. We need to develop educational methods that cultivate a well-balanced clinical sense to enable students to choose a suitable diagnostic strategy as needed in a clinical setting without being one-sided to the "rule-in conscious paradigm." PMID- 15109341 TI - All dysuria is local. A cost-effectiveness model for designing site-specific management algorithms. AB - OBJECTIVE: To determine the cost-effectiveness of management strategies for dysuria in different office settings. DESIGN: Decision and cost-effectiveness analyses, assuming the payer's perspective. Data on disease prevalence, test characteristics, treatment efficacy, and adverse effects were drawn from the English language literature using medline searches and bibliographies. SETTING: Hypothetical primary care practice. PATIENTS: Otherwise healthy, nonpregnant women with symptoms of dysuria, urgency, and frequency. INTERVENTIONS: All reasonable combinations of urinalysis, urine culture, pelvic examination, chlamydia and gonorrhea cultures, and empiric treatment with trimethoprim sulfamethoxazole. RESULTS: The cost-effectiveness of strategies varied substantially among different patient settings. In all settings, empiric trimethoprim-sulfamethoxazole for all patients was least expensive and least effective. Most testing increased both cost and effectiveness. Compared to empiric antibiotics, performing pelvic examination and urine culture for women with normal urinalyses had a marginal cost-effectiveness ratio of $4 to $32 per symptom-day avoided (SDA). Adding urine culture for patients with pyuria had a marginal cost of $34 to $107 per SDA, which fell to $40/SDA when the prevalence of resistance to trimethoprim-sulfamethoxazole exceeded 40%. Pelvic examination and urine culture for all patients regardless of urinalysis results achieved the greatest benefit but at the highest cost (>$300 per SDA). CONCLUSIONS: In otherwise healthy women with symptoms of dysuria and no vaginal complaints, performing pelvic exam and urine culture based on urinalysis offers a reasonable alternative to empiric therapy. Other testing may be warranted, depending on antibiotic resistance and the value of avoiding a day of dysuria. PMID- 15109342 TI - Consistency of performance ranking of comorbidity adjustment scores in Canadian and U.S. utilization data. AB - OBJECTIVE: The performance of standard comorbidity scores to control confounding is poorly defined in health care utilization data across elderly populations. We sought to evaluate and rank the performance of comorbidity scores across selected U.S. and Canadian elderly populations using health care utilization databases. DESIGN: Cross-population validation study. PARTICIPANTS: Study participants were residents age 65 years or older who had prescription drug coverage through state funded programs selected from several large health care utilization databases available to the investigators: British Columbia, BC (N = 141,161), New Jersey, NJ (N = 235,881), and Pennsylvania, PA (N = 230,913). MEASUREMENTS: We calculated 6 commonly used comorbidity scores for all subjects during the baseline year (1994 for NJ and PA, and 1995 for BC). These included scores based on diagnoses (Romano, Deyo, D'Hoore, Ghali) and prescription drugs (CDS-1, CDS-2). The study outcome was 1-year mortality. The performance of scores was measured by c statistics derived from multivariate logistic regression that included age and gender. MAIN RESULTS: Across these 4 large elderly populations, we found the same rank order of performance in predicting 1-year mortality after including age and gender in each model: Romano (c-statistic 0.754 to 0.771), Deyo (c-statistic 0.753 to 0.768), D'Hoore (c-statistic 0.745 to 0.760), Ghali (c-statistic 0.733 to 0.745), CDS-1 (c-statistic 0.689 to 0.738), CDS-2 (c-statistic 0.677 to 0.718), and age and gender alone (c-statistic 0.664 to 0.681). Performance was improved by an average of 6% by adding the number of different prescription drugs received during the past year. CONCLUSIONS: Performance ranking of 6 frequently used comorbidity scores was consistent across selected elderly populations. We recommend that investigators use these performance data as one important factor when selecting a comorbidity score for epidemiologic analyses of health care utilization data. PMID- 15109343 TI - Differential willingness to undergo smallpox vaccination among African-American and white individuals. AB - OBJECTIVE: To examine potential disparities in willingness to be vaccinated against smallpox among different U.S. racial/ethnic groups. DESIGN: Cross sectional survey using an experimental design to assess willingness to be vaccinated among African Americans compared to whites according to 2 strategies: a post-exposure "ring vaccination" method and a pre-exposure national vaccination program. SETTING: Philadelphia County district courthouse. PARTICIPANTS: Individuals awaiting jury duty. MEASUREMENTS: We included 2 scenarios representing these strategies in 2 otherwise identical questionnaires and randomly assigned them to participants. We compared responses by African Americans and whites. MAIN RESULTS: In the pre-exposure scenario, 66% of 190 participants were willing to get vaccinated against smallpox. In contrast, 84% of 200 participants were willing to get vaccinated in the post-exposure scenario (P =.0001). African Americans were less willing than whites to get vaccinated in the pre-exposure scenario (54% vs 77%; P =.004), but not in the post-exposure scenario (84% vs 88%; P =.56). In multivariate analyses, overall willingness to undergo vaccination was associated with vaccination strategy (odds ratio, 3.29; 95% confidence interval, 1.8 to 6.1). CONCLUSIONS: Racial disparity in willingness to get vaccinated varies by the characteristics of the vaccination program. Overall willingness was highest in the context of a post-exposure scenario. These results highlight the importance of considering social issues when constructing bioterror attack response plans that adequately address the needs of all of society's members. PMID- 15109344 TI - Determinants of resident satisfaction with patients in their continuity clinic. AB - The purpose of this study was to identify what patient and physician factors influence resident satisfaction with patient encounters in a continuity clinic setting. Resident satisfaction was assessed from postencounter questionnaires completed by 68 internal medicine residents regarding 979 patient encounters. We found that residents were more satisfied with patients diagnosed with general medical problems than with patients diagnosed with pain and psychiatric disorders. First-year residents were less satisfied with patients diagnosed with pain and psychiatric disorders than second- and third-year residents. However, this dissatisfaction with seeing patients with pain or psychiatric disorders lessened as continuity of care was enhanced. PMID- 15109345 TI - The use of "overall accuracy" to evaluate the validity of screening or diagnostic tests. AB - OBJECTIVE: Evaluations of screening or diagnostic tests sometimes incorporate measures of overall accuracy, diagnostic accuracy, or test efficiency. These terms refer to a single summary measurement calculated from 2 x 2 contingency tables that is the overall probability that a patient will be correctly classified by a screening or diagnostic test. We assessed the value of overall accuracy in studies of test validity, a topic that has not received adequate emphasis in the clinical literature. DESIGN: Guided by previous reports, we summarize the issues concerning the use of overall accuracy. To document its use in contemporary studies, a search was performed for test evaluation studies published in the clinical literature from 2000 to 2002 in which overall accuracy derived from a 2 x 2 contingency table was reported. MEASUREMENTS AND MAIN RESULTS: Overall accuracy is the weighted average of a test's sensitivity and specificity, where sensitivity is weighted by prevalence and specificity is weighted by the complement of prevalence. Overall accuracy becomes particularly problematic as a measure of validity as 1) the difference between sensitivity and specificity increases and/or 2) the prevalence deviates away from 50%. Both situations lead to an increasing deviation between overall accuracy and either sensitivity or specificity. A summary of results from published studies (N = 25) illustrated that the prevalence-dependent nature of overall accuracy has potentially negative consequences that can lead to a distorted impression of the validity of a screening or diagnostic test. CONCLUSIONS: Despite the intuitive appeal of overall accuracy as a single measure of test validity, its dependence on prevalence renders it inferior to the careful and balanced consideration of sensitivity and specificity. PMID- 15109346 TI - Inpatient diabetology. The new frontier. AB - Tight glycemic control is now an imperative of outpatient diabetes care. The inpatient arena remains under the influence of an ineffective paradigm characterized by tolerance for hyperglycemia and a reluctance to use insulin intensively. This article is a call to action against the lip service paid to inpatient diabetes care. The compelling in vitro and in vivo evidence for the benefit of intensive insulin-mediated glycemic control is summarized. The linchpin of current inpatient care is a commonly used insulin sliding scale. This autopilot approach as the sole mode of treatment for inpatient hyperglycemia has been strongly condemned. Nevertheless, it continues to survive. The evidence supports the compelling argument that the adverse effect of hyperglycemia on hospital length of stay, morbidity, and mortality is substantial. Clinicians, nurses, administrators, and insurers ought to look critically at the prevailing paradigm and spearhead the much-needed revolution in inpatient diabetology. The issue of glycemic targets, the need for noninvasive blood glucose monitoring, and the role of nursing staff in this revolution are raised. We call for the banning of the insulin sliding scale use as the sole diabetes order. Also, the use of basal insulin via continuous intravenous insulin infusion or subcutaneous insulin analogs should be embraced. Educating nurses, house staff, and other frontline professionals in the adverse consequences of the current paradigm is essential. Inpatient glycemic control matters; clinical and financial outcomes are at stake. It behooves the health care system and the diabetic public to address the contemporary state of inpatient diabetology as soon as possible. PMID- 15109347 TI - Getting funded. Career development awards for aspiring clinical investigators. AB - For aspiring clinical investigators, career development awards provide a primary mechanism for "getting funded." The objective of this article is to provide information that will facilitate a successful application for a research career development award. Specifically, we discuss important issues that cut across the diverse array of awards, and we highlight the most common sources of funding, including the unique opportunities that are available for underrepresented minorities. The target audience includes junior faculty and fellows who are pursuing or considering a research career in academic medicine, as well as their mentors and program directors. PMID- 15109348 TI - Hospital care of general medicine patients: the importance of evidence. PMID- 15109349 TI - Selling the "quit" brand to young adult smokers. PMID- 15109353 TI - Sympathetic skin response in migraineurs and patients with medication overuse headache. AB - BACKGROUND: Autonomic dysfunction has been reported in patients with migraine, and it may play a role in promoting attacks. OBJECTIVE: To investigate changes in the autonomic function of migraineurs and patients with medication overuse headache via sympathetic skin response, and to determine whether psychiatric comorbidity is related to any changes recorded. METHODS: A consecutive series of patients with migraine (n = 45) and medication overuse headache (n = 53) were studied. Patients with other chronic diseases requiring medication were excluded. Sympathetic skin response latencies and amplitudes from the patients with headache (N = 98) and 40 healthy controls were compared statistically. RESULTS: Sympathetic skin response latencies in patients with medication overuse headache and in migraineurs were significantly longer than in controls. To analyze the effect of psychiatric comorbidity, patients with medication overuse headache and migraineurs were each divided into 2 groups: those with psychiatric comorbidity and those without comorbidity. When the sympathetic skin response results of these 4 groups were compared with controls, the only statistically significant difference was between the sympathetic skin response latencies of controls and the latencies of patients with psychiatric comorbidity. We could not find any difference between the results from patients without psychiatric comorbidity and those of controls. CONCLUSION: Psychiatric disease may affect the results of autonomic function testing in migraineurs and patients with medication overuse headache. Consideration should be given to excluding patients with psychiatric comorbidity from studies investigating autonomic dysfunction in patients with headache. PMID- 15109354 TI - Familial migraine with aura: association study with 5-HT1B/1D, 5-HT2C, and hSERT polymorphisms. AB - BACKGROUND: The serotonergic system has a significant role in the pathophysiology and pharmacology of migraine. OBJECTIVE: To study the association between the occurrence of migraine with aura and 5-HT(1B/1D) and 5-HT(2C) receptor gene and the human serotonin transporter (hSERT) gene polymorphisms in 18 unrelated families with multiple affected members. METHOD: Two polymorphisms in the 5 HT(1B/1D) receptor gene and one polymorphism in the 5-HT(2C) receptor gene were studied by restriction fragment length polymorphism analysis. Allelic variation of the hSERT, with 9, 10, and 12 copies of a "repetitive element," was studied by polymerase chain reaction amplification of the variable number tandem repeat region. RESULTS: Allelic distribution of 5-HT(1B/1D) and 5-HT(2C) receptor gene polymorphisms in affected patients did not differ in either of the control groups (unaffected relatives or unrelated healthy individuals). A trend toward a significant effect of the 12-repeat hSERT allele as a risk factor for migraine with aura versus unrelated controls was observed. CONCLUSION: Our data do not support the involvement of 5-HT(1B/1D) and 5-HT(2C) receptor gene polymorphisms in migraine with aura, yet do suggest a possible role for a locus at or near the hSERT gene in the susceptibility to migraine with aura. PMID- 15109355 TI - Almotriptan improves response rates when treatment is within 1 hour of migraine onset. AB - BACKGROUND: Results from open-label trials with almotriptan and sumatriptan have shown higher response rates when treatment was initiated early after acute migraine onset. OBJECTIVE: To investigate the temporal component of early intervention by measuring 2-hour pain-free and sustained pain-free responses to almotriptan and sumatriptan when the study drug was taken within 1 hour of onset of moderate to severe pain. METHODS: This was a post hoc analysis from a double blind, randomized, placebo-controlled trial of almotriptan and sumatriptan. Men and women, 18 to 65 years of age, who met International Headache Society criteria for migraine with or without aura were eligible. Patients were randomized to receive a single oral dose of almotriptan 12.5 or 25 mg, sumatriptan 100 mg, or placebo at the onset of a severe or moderate migraine attack. For this post hoc analysis, the almotriptan 25-mg dose was excluded because 12.5 mg is the recommended dose. The primary efficacy assessment was sustained pain-free, defined as pain-free at 2 hours postdose with no recurrence from 2 to 24 hours and no use of rescue medication. Only patients who took study medication within 1 hour of migraine onset were included in the analysis. RESULTS: Of the 475 patients involved in the original study, 253 (53.3%) initiated treatment within the 0- to 1-hour interval. For these patients, 2-hour pain-free rates were 37.9% for almotriptan 12.5 mg (P=.016 versus placebo), 35.7% for sumatriptan 100 mg (P=.028 versus placebo), and 18.9% for placebo. Only almotriptan was significantly higher than placebo on the sustained pain-free rate-34.7% (P=.022 versus placebo); the sustained pain-free rate for sumatriptan was 29.6% and for placebo, 17.0%. CONCLUSION: Initiation of treatment with almotriptan 12.5 mg within the first hour after acute migraine onset resulted in a significantly higher sustained pain-free response compared with placebo. There was no significant difference in sustained pain-free rates between sumatriptan and placebo. These results are consistent with those from a previous open-label trial, and suggest that early intervention with almotriptan can improve clinical outcome. PMID- 15109356 TI - Simple and efficient recognition of migraine with 3-question headache screen. AB - OBJECTIVE: To correlate the results of a new 3-question headache screen to 3 established methods of diagnosing migraine: the International Headache Society diagnostic criteria, physician's clinical impression, and presence of recurring disabling headaches. BACKGROUND: A simple tool to recognize patients who experience migraine may facilitate diagnosis of this debilitating and frequently undiagnosed condition. METHODS: Primary care physicians and neurologists in the United States enrolled 3014 adults with a diagnosis of migraine based on one of the following: International Headache Society criteria, an investigator's clinical impression, or presence of recurring disabling headaches. Each patient completed a 3-question headache screen: (1) Do you have recurrent headaches that interfere with work, family, or social functions? (2) Do your headaches last at least 4 hours? (3) Have you had new or different headaches in the past 6 months? A diagnosis of migraine was suggested by a yes answer to questions 1 and 2 and a no answer to question 3. RESULTS: The 3-question headache screen identified migraine in 77% of the study population; including 78% of the patients enrolled based on International Headache Society criteria, 74% based on clinical impression, and 68% because of recurring disabling headaches. CONCLUSIONS: Positive 3-question headache screen results agreed well with migraine diagnoses based on International Headache Society criteria, clinical impressions, and presence of recurring disabling headaches. These findings support use of the 3 question headache screen to recognize migraine. PMID- 15109357 TI - Validation of a brief nurse-administered migraine assessment tool. AB - OBJECTIVE: To validate a brief tool for screening migraine. BACKGROUND: Migraine is a common, but underdiagnosed condition. Effective utilization of nonphysician personnel to reliably screen patients for migraine may improve identification of migraineurs for clinical treatment and research. METHODS: An 8-question Migraine Assessment Tool (based on International Headache Society criteria) was designed for administration by a nurse with no specialized headache training as a pre assessment for the diagnosis of migraine for use in either a research or clinical environment. A community sample of 80 adults (71 women, 9 men; mean age, 33.7 years; 80% white, 14% African American, 2.5% Asian American) with self-reported headache was recruited through advertisements. A headache specialist independently diagnosed subjects using clinical assessment, and a nurse who works in a balance disorder clinic used the Migraine Assessment Tool. Agreement between physician and nurse-administered Migraine Assessment Tool diagnoses was determined. Each subject returned in 2 to 4 weeks for a second assessment, administered by the same nurse. Agreement between the 2 diagnoses from the Migraine Assessment Tool was calculated. RESULTS: Comparison between diagnosis by the physician versus the Migraine Assessment Tool revealed a positive predictive value of 0.85; negative predictive value, 0.84; sensitivity, 0.89; specificity, 0.79; and observed agreement, 0.85. Cohen's kappa reliability measure was 0.69, indicating good test reliability. Interestingly, in 8 of the 12 cases of disagreement, the examiner diagnosing nonmigraine diagnosed analgesic overuse headache. Comparing diagnoses assigned by the 2 separate administrations of the Migraine Assessment Tool revealed a Cohen's kappa of 0.69. Notably, 9 of the 12 cases of nonagreement on the 2 assessments were due to subjects endorsing analgesic overuse in only 1 of the 2 testing sessions. CONCLUSIONS: This study showed good reliability and stability of a new, brief, nurse-administered migraine questionnaire. In addition, the study also showed that consistency in self-reporting analgesic overuse within individuals with headache is poor. This suggests the need for repeat questioning about analgesic overuse on subsequent appointments to ensure absence of analgesic overuse headache. PMID- 15109358 TI - Electroacupuncture for tension-type headache on distal acupoints only: a randomized, controlled, crossover trial. AB - OBJECTIVE: To investigate the efficacy of electroacupuncture, applied to distal acupoints only, for tension-type headache. BACKGROUND: Electroacupuncture is commonly used for tension-type headache, but when applied to distal acupoints only, evidence of its efficacy is lacking. DESIGN: A randomized, single-blinded, sham-controlled, crossover clinical trial. Methods.-The trial had 5 stages: baseline (2 weeks), phases I and II (each 4 weeks), washout period (2 weeks), and follow-up (3 months after phase II). Forty patients were randomly assigned to either group A or group B. Group A received real electroacupuncture during phase I, then sham electroacupuncture in phase II. Group B received the treatments in reverse order. Outcome measures were headache frequency and duration, pain intensity using a visual analog scale, mechanical pain threshold, headache disability, and sickness impact. Data were analyzed by univariate 2-way analysis of variance. RESULTS: Thirty-seven patients completed the trial. There were no significant differences between the 2 groups at baseline. At the end of phase I, group A, but not group B, demonstrated significant improvement in mean (standard error of the mean [SEM]) headache frequency (3.0 per month [0.3] versus 12.0 per month [1.7]), duration (13.3 hours [3.5] versus 32.0 hours [6.2]), pain intensity (32.8 mm [4.1] versus 47.5 mm [2.7]), pain threshold (right side, 2.9 kg/second [0.1] versus 0.9 kg/second [0.1]; left side, 2.4 kg/second [0.1] versus 1.1 kg/second [0.1]), headache disability score (6.0 [1.0] versus 16.3 [1.6]), and sickness impact score (288.7 [48.0] versus 687.1 [77.2]). For each parameter, significant differences also were demonstrated for both groups between baseline and phase II, and baseline and follow-up. There were no significant differences between the groups at the end of follow-up (P >.05). CONCLUSION: Electroacupuncture to distal points alone is effective for short-term symptomatic relief of tension-type headache. PMID- 15109359 TI - Inpatient treatment of headache: an evidence-based assessment. AB - OBJECTIVE: To evaluate inpatient treatment of headache in the United States. PARTICIPANTS: Participants were selected by the meeting chairpersons, Drs. Diamond and Silberstein as well as Dr. Freitag. Criteria for selection included participation in an inpatient treatment program, headache specialists not affiliated with an inpatient treatment program, clinician with interest in headache but not affiliated with a dedicated headache treatment program. The single meeting held in New York was by invitation only. All invitees were provided with opportunity to present their views and participate in the recommendations of the Consortium. The list of participants at the meeting were: Harvey Blumenthal, MD, Roger Cady, MD, James Couch, MD, Seymour Diamond, MD, Frederick G. Freitag, DO, R. Michael Gallagher, DO, Al Lake III, PhD, Richard Lipton, MD, NinanMathew,MD, Alan Rapoport, MD, Jay Rosenberg, MD, Joel Saper, MD, Stephen Silberstein, MD. Funding for theproject was provided by the US Headache Guidelines Consortium Project through the American Academy of Neurology and the National Headache Foundation. EVIDENCE: Each of the authors were part of the writing committee each with specific areas that they were responsible for conducting the literature reviews and submitting a portion of the original document for editing and review. Dr. Freitag coordinated this process and assembled the first draft of the article for circulation and review among the authors and chairpersons. In the absence of class I evidence consensus expert opinion was fundamental to the process. CONCLUSIONS: The treatment of headache is commonly delivered in the outpatient office environment. There exists a group of patients who have been refractive to outpatient treatment and in whom in patient care may be appropriate. The factors that determine the optimal treatment setting, the methodology of the treatment and the results of such treatment have come under increasing scrutiny secondary to the cost of treatment in the hospital. The US Headache Consortium guidelines project in conjunction with the national headache foundation believed that the need existed to objectively examine the issues surrounding inpatient treatment. The epidemiology of chronic headache is important to understand in this context since essentially all patients that are treated in hospital have high frequency headache often associated with significant disability. This severity of illness produces both health care costs but also is responsible for significant impact on businesses and the economy. The method of treatment of headache in the hospital environment can follow several paths based on the accessibility of the resources specific to headache treatment. While there exist these differing paths of treatment only treatment in a dedicated headache treatment unit has been investigated in depth here in the US though reports from other countries that do not have dedicated inpatient treatment lends support to options in headache treatment. Some have suggested that inpatient treatment is not required even in recidivist patients and limited reports have examined aggressive outpatient treatment as an option in headache management. None of the studies reported to date provide class one evidence for efficacy either inpatient or outpatient. Recommendations are made to assess headache treatment in patients with high frequency headaches to garner better scientific evidence for differing treatment approaches. PMID- 15109360 TI - Eletriptan for the short-term prophylaxis of cluster headache. AB - BACKGROUND: A beneficial prophylactic effect from eletriptan 40 mg given to a single patient with cluster headache was observed. OBJECTIVE: To further evaluate the efficacy of eletriptan in the short-term prophylaxis of cluster headache. METHODS: We treated 18 patients; mean age, 40.5 years (standard deviation [SD], 9.9). The number of cluster headache attacks was recorded during a baseline period of 6 days, and during 6 days of treatment with eletriptan 40 mg twice daily. The primary outcome measure was the reduction in the number of attacks during the treatment period. RESULTS: In the 16 patients who completed the study (2 patients were lost to follow-up), the mean total number of attacks decreased from 10.9 (SD, 5.6) during baseline to 6.3 (SD, 3.7) during treatment with eletriptan (P=.01) The reduction in the number of attacks exceeded 50% in 6 patients. CONCLUSION: This small open-label study suggests that eletriptan 40 mg twice daily may be useful for the short-term prophylaxis of cluster headache. PMID- 15109361 TI - Prednisolone-responsive headache in patients with solitary cysticercus granuloma and seizures. AB - BACKGROUND: Solitary cysticercus granuloma is the commonest imaging abnormality in Indian patients with new-onset seizures. Few patients, in addition, complain of disabling headache. OBJECTIVE: To report our experience of 16 patients with new-onset headache, seizures, and solitary cysticercus granuloma. METHODS: Sixteen consecutive patients, who had moderate to severe new-onset headache and fulfilled the diagnostic criteria of solitary cysticercus granuloma, were included in the study. The inclusion criteria were: occurrence of seizures, minimal or no neurologic deficit, absence of papilledema, no evidence of any systemic disorder, and computed tomography showing a single ring/disk-enhancing lesion of <20 mm in diameter. Patients received antiepileptic monotherapy, oral analgesics, and prednisolone (1.5 mg/kg/day for 7 days). Prednisolone was then tapered over the next 7 days. Patients were followed for 6 months. Follow-up computed tomography was performed after 2 months; in all 16 patients, the scans showed complete disappearance of the lesion. RESULTS: After 14 days (at first follow-up), all patients reported significant improvement in headache. Follow-up after 2 months revealed that all patients were headache-free. None of the patients reported any recurrence of headache. CONCLUSION: This open-label study suggests the effectiveness of prednisolone for disabling headache in Indian patients with solitary cysticercus granuloma and seizures. There is a need, however, for more scientifically rigorous studies for further confirmation of our results. PMID- 15109362 TI - Septoplasty for migraine? PMID- 15109363 TI - Tapeworm headache. PMID- 15109364 TI - Choto-san (kampo Medicine) for the treatment of headache. PMID- 15109365 TI - De novo headache and analgesic consumption: pathophysiological insights from nosologic complexity? PMID- 15109375 TI - Rectal cancer: improving outcomes. PMID- 15109376 TI - Pouchitis. AB - Pouchitis is one of the commonest and most debilitating complications of a restorative proctocolectomy. The cause remains elusive, though a number of approaches have been shown to alleviate the condition. This review outlines current evidence relating to pouchitis, obtained from randomised and nonrandomised studies. Medline, the Bath Information Data Service (BIDS) and PubMed were searched using the keywords 'pouchitis' and 'inflammatory bowel disease'. In addition, articles were cross-referenced, and the abstracts of recent colorectal meetings studied. PMID- 15109377 TI - Survival of rectal cancer patients in Denmark during 1994-99. AB - OBJECTIVE: As survival from rectal cancer in Denmark is below the European average, we analysed survival during the period of 1994-99 focusing upon improvement strategies. METHOD: All patients with a first-time rectal cancer were registered in a national database during this 5-year period. In the observational cohort study, data on patient age and gender, tumour stage, surgical procedures, adjuvant radiotherapy, anastomotic leakage, 30-day mortality and long-term survival were evaluated. RESULTS: The database comprised 5021 patients. Sixty four percent had a localized tumour. Less than a third of patients with fixed tumours had pre-operative radiotherapy and curative surgery was achieved in 70%. Anastomotic leakage occurred in 13%, and 30-day mortality was 4% following abdominoperineal or anterior resection and 11% following a Hartmann's procedure. The relative 5-year survival in the entire series was 39% in males and 47% in females, respectively. Following curative surgery the relative 5-year survival was 55% in males and 63% in females, respectively. Survival was 71% in the subset of patients receiving curative total mesorectal excision. CONCLUSION: The average tumour stage upon diagnosis was probably more advanced compared to the other Nordic countries and pre-operative radiotherapy was administered to a minority of patients with fixed tumours. The anastomotic leakage rate was relatively high, whereas the 30-day mortality was comparable to other studies. Survival from rectal cancer in Denmark is still less favourable compared to the other Nordic and several European countries but improved from 1996 and onwards. PMID- 15109378 TI - High compliance rates observed for follow up colonoscopy post polypectomy are achievable outside of clinical trials: efficacy of polypectomy is not reduced by low compliance for follow up. AB - INTRODUCTION: The National Polyp Study demonstrated that removal of adenomas with at least a three-year follow up reduced the incidence of colorectal cancer. However, compliance with follow up colonoscopy may affect the estimates of reduction in colorectal cancer incidence demonstrated by the National Polyp Study. While an 80% compliance rate for follow up colonoscopy was achieved during the National Polyp Study, the compliance rate for follow up colonoscopy is unknown in the general population. The aim of this study was to determine the compliance rate for follow up colonoscopy and factors which affect follow up. METHODS: A retrospective medical record review to identify patients who had adenomatous polyps excised in 1997 was undertaken. Patients who had inflammatory bowel disease, a prior history of colorectal cancer, familial adenomatous polyposis syndrome, colonic surgery; incomplete polypectomy or incomplete colonoscopy, and those patients who died before planned follow up, were excluded from analysis. Follow up was performed by telephone survey. RESULTS: Three hundred and thirty-three patients were identified (196 males; 147 females) with a mean age of 70 years. Three hundred and thirty-one (99%) of 333 had a documented recommendation for follow up of three years or less. Thirty-four percent (113 of 333) had previously undergone colonoscopy; 29% (98 of 333) had previously undergone polypectomy and 54% (180 of 333) were symptomatic at the time of the colonoscopy. Twenty-eight percent (40 of 141) had a family history of colorectal cancer. Pathology at polypectomy included a single polyp and polyps less than 10 mm in 68% and 88% of cases, respectively. Follow up was available in 211 of these cases, 179 (85%) of which had been compliant with follow up colonoscopy. In a univariate analysis, previous colonoscopy (P = 0.035), previous polyps (P = 0.043), asymptomatic status at time of colonoscopy (P = 0.021), polyp size (P = 0.008) and number of polyps (P = 0.010) were significantly associated with patients who were compliant with follow up colonoscopy. A multivariate logistics regression analysis revealed number of polyps (P = 0.036) and polyp size (P = 0.045) to be statistically significantly associated with compliance. CONCLUSION: Compliance with follow up colonoscopy after polypectomy is greater than 80%, regardless of age, education, family history, prior colonoscopy, or prior polypectomy. Risk reduction published in the National Polyp Study may likely reflect what can be achieved through the general use of colonoscopy for surveillance. PMID- 15109379 TI - Colonic pouchography is not routinely required prior to stoma closure. AB - BACKGROUND: Colonic pouch formation with pouch-anal anastomosis is now regarded as the procedure of choice for restoration of intestinal continuity following anterior resection for low rectal cancers. The aim of this study was to review the necessity for routine colonic pouchography prior to closure of a diverting loop stoma. METHODS: This was a prospective study of 52 consecutive patients who underwent colonic pouch formation between 1 June 1999 and 31 May 2002, four of whom have subsequently died. Each pouch was assessed clinically and radiologically prior to stoma closure. RESULTS: There were no clinical anastomotic leaks. Forty-six of 48 surviving patients have had a colonic pouchogram and in no case was either a pouch or pouch-anal anastomotic defect identified. To date 40 patients have undergone stoma closure without an anastomosis-related complication. CONCLUSION: Following successful colonic pouch formation, routine study of the pouch by contrast radiology does not add to clinical assessment. As a consequence radiological imaging is unnecessary and can be omitted. PMID- 15109380 TI - Abdominal stapled side-to-end anastomosis (Baker type) in low and high anterior resection: experiences and results in 69 consecutive patients at a regional general hospital in Japan. AB - OBJECTIVE: The technique of transanally introducing a circular stapled device to accomplish colorectal anastomoses has been widely used. However, the widespread popularity of this technique may have created the potential of anal sphincter injury during transanal insertion of the anastomosing stapler. Thus, to avoid the risk of anal sphincter injury during anal manipulation, we have been performing an abdominal approach, namely abdominal stapled side-to-end anastomosis (ASSEA) using a Purstring and premium curved EEA stapler in low and high anterior resection. This study will present our experience and results of consecutive resections. METHODS: ASSEA following a resection of the rectum and sigmoid colon for carcinomas was consecutively performed between October 1998 and May 2002. Age, sex, pre-operative anal function, the TNM classification by the UICC rules, operative methods, postoperative morbidity, mortality, anal function and bowel frequency were evaluated. RESULTS: Sixty-nine consecutive patients underwent a resection of the rectum and sigmoid colon with ASSEA. There were three (4.3%) clinical anastomotic leakages in the cases studied. Anastomoses that were located above the peritoneal reflection leaked in one (2.5%) of 40 cases, while anastomoses below the peritoneal reflection leaked in two (6.9%) of 29 cases. A diverting stoma was performed in five (17%) of the 29 cases with low anastomosis. Postoperative anal function was stable without soiling or faecal leakage and bowel frequency two months after surgery was less than four times in all 61 patients. There was no postoperative mortality related to the anastomosis. CONCLUSIONS: Abdominal stapled side-to-end anastomosis (Baker type) was found to be a safe and relatively easy method in both low and high anterior resection in association with a good quality of life. PMID- 15109381 TI - The effect of diverticular disease on the colonic J pouch. AB - OBJECTIVE: The aim of this study was to assess the impact of the diverticular disease (DD) on function and on postoperative complications of the colonic J pouch (CJP) with pouch-anal anastomosis. METHODS: Patients who underwent a CJP between December 1990 and August 2001, were retrospectively reviewed. The presence of DD in the CJP was assessed on pouchogram prior to ileostomy closure. A questionnaire designed to evaluate the degree of continence (total incontinence score (IS): 0 = worst, 20 = best) and pouch evacuation (total evacuation score (ES): 0 = worst, 28 = best) was used for comparison between patients with DD and those without DD (NDD). RESULTS: Sixty-six patients (47 males; 19 females) with a median age of 68 years (range 28-87 years) were included. The median follow-up period was 22 months (range 2-106 months). Twenty-four patients comprised the DD group and 42 were in the NDD group. The two groups were comparable for age, gender and time from ileostomy closure; all patients with postoperative chemoradiation therapy were in the NDD group. The total ES and IS total did not significantly differ between the two groups with a P-value of 0.11 and 0.09 respectively. Furthermore, there was no significant difference in the total incidence of pouch complications between the two groups (3 strictures, 1 leak, 1 fistula in the NDD group vs. 1 pelvic sepsis in the DD group; P = 0.4). CONCLUSIONS: The presence of DD in a CJP does not seem to impact pouch function or the postoperative complication rate. PMID- 15109382 TI - Surgery for occult rectal prolapse. AB - OBJECTIVE: An 'occult' rectal prolapse may be diagnosed during investigation of altered bowel habit. It has been suggested that the outcome of surgery for these patients may be associated with results that are inferior to those achieved in patients with overt rectal prolapse. This study compares the results of surgery for 'occult' and overt rectal prolapse in terms of mortality, morbidity and change in bowel habit. PATIENTS AND METHODS: A retrospective review was undertaken of consecutive patients undergoing surgery for rectal prolapse during the decade 1988-98. Resection rectopexy was the treatment of choice except in patients with faecal incontinence who underwent sutured rectopexy. Those patients who were unfit for an abdominal operation were offered a perineal procedure. Outcome measures were mortality, morbidity, prolapse recurrence, constipation and faecal incontinence. Data were retrieved from case note review, clinical assessment, telephone consultation or postal questionnaire. RESULTS: Rectal prolapse surgery was undertaken in 69 patients with an overt prolapse and 74 patients with an 'occult' prolapse. Patients in the 'occult' prolapse group were significantly younger than those with overt prolapse (P = 0.0002). There were significantly more perineal procedures in the overt prolapse group compared with the 'occult' prolapse group (54% vs 5%, P = 0.0001). There were no deaths within 28 days of surgery. Major surgical complications occurred in 5 patients (3.5%). Seven patients (10%) experienced recurrent prolapse. Rectal prolapse surgery reduced the incidence of St. Mark's grade 4 faecal incontinence from 38% to 19% in the overt prolapse group (P = 0.023) and from 49% to 22% in the 'occult' prolapse group (P < 0.001). Following surgery the incidence of constipation increased in the 'occult' group from 39% to 50% but decreased in the overt prolapse group from 42% to 35%. CONCLUSIONS: Surgery for an 'occult' rectal prolapse is unlikely to benefit patients whose principle symptom is constipation. Approximately half of those patients whose 'occult' rectal prolapse is associated with faecal incontinence will have their bowel habit improved by prolapse surgery. PMID- 15109383 TI - Artificial bowel sphincter in severe anal incontinence. AB - OBJECTIVE: The artificial anal sphincter has been suggested as an alternative in the treatment of severe anal incontinence when conventional surgical methods are not possible or have failed. Experience in this procedure is still limited and the results have not yet been sufficiently established. The aim of this study is to evaluate the efficacy of the ACTICON (American Medical Systems, Minneapolis, MN) on patients operated upon in our Unit. PATIENTS AND METHOD: In this prospective study an ACTICON sphincter was implanted in 10 patients (8 women) with an average age of 56 years and with an average period of severe anal incontinence of 151 months. The origin of incontinence was obstetric injury (n: 4), neuropathy (n: 3) and sphincteral injury from previous anal surgery (n: 3). The degree of continence was measured using the Fecal Incontinence Scoring System (FISS) and the pre- and postoperative anal manometric parameters at 6-month intervals. The average follow-up time for the efficacy of the implanted system was 29 months. RESULTS: A total of 6 patients [60%] displayed complications in the immediate postoperative period: subaponeurotic reimplantation of the connecting tubes was necessary after infection of the abdominal wound (n:1); superficial dehiscence of the perianal wound (n: 2), infection of the perianal wound (n: 1) and perianal haematoma (n: 2) that were resolved by conservative treatment. For 3 patients [30%] the system was explanted, definitively in one and in 2 of them reimplanted successfully. At the end of the follow-up period, 9 patients [90%] still have an activated artificial sphincter. The score on the Fecal Incontinence System decreased significantly after the system was activated (P < 0.0001) and the pressure with the cuff closed was significantly higher than pre-operative anal pressure (P < 0.0001). All the patients are now continent for solid stool, 56% have occasional involuntary losses of gases and 33% occasionally have involuntary losses of gases and liquid stool. Only 2 patients [22%] have complete continence. CONCLUSIONS: Our findings indicate that the ACTICON artificial anal sphincter is well tolerated and can be an effective alternative in the treatment of severe anal incontinence. Although complete continence is only achieved in a low percentage of cases, for the rest of the patients the ACTICON neosphincter reduces the symptoms considerably. PMID- 15109384 TI - A retrospective evaluation of rectal irrigation in the treatment of disorders of faecal continence. AB - OBJECTIVES: The aim of this study was, firstly, to evaluate the efficacy and acceptability of rectal irrigation in the treatment of patients with disorders of faecal continence that have not responded to other treatment modalities. Secondly, to see if standard anorectal physiology measures can identify patients who might have successful treatment with rectal irrigation. PATIENTS AND METHODS: All patients who had been treated with rectal irrigation, between 1998 and 2000, were sent a postal questionnaire. Patients were asked to quantify their symptoms (before and after rectal irrigation) using a visual analogue scale to determine the efficacy of rectal irrigation. An increase of 10 (10 mm) in this score was regarded as successful treatment and incontinence scores obtained. The acceptability of rectal irrigation was determined using a Quality of Life questionnaire. Data regarding presenting symptoms, previous therapies, pretreatment anorectal physiology measurements were obtained from a retrospective case note and database review. RESULTS: Completed questionnaires were obtained from 48 (52%) of 92 patients, 39 patients had had previous medical and/or surgical treatment before trying rectal irrigation. At the time of the questionnaire 44 of 48 patients were still using rectal irrigation. Using the linear analogue scores 24 of 48 (50%) reported an improvement in their symptoms using rectal irrigation. Most patients found the treatment acceptable. Incontinence scores and anorectal physiology measures did not predict those patients who responded successfully to rectal irrigation. CONCLUSION: Rectal irrigation can offer symptomatic improvement to patients with faecal evacuatory disorders where other therapies have failed. Most patients find the treatment acceptable. Standard anorectal physiology measures and incontinence scores do not predict those patients likely to have successful treatment. PMID- 15109385 TI - Colorectal cancer in the young: a 12-year review of patients 30 years or less. AB - OBJECTIVES: As the incidence of young colorectal cancer is rising, a review of the characteristics of such malignancy in those under 30 years of age is timely at this stage. PATIENTS AND METHODS: Thirty-nine patients (21 M, 18 F) were operated upon over a 12-year period in a single centre. The mean age was 25 years and median follow-up was 20 months. RESULTS: Rectal bleeding, change in bowel habit and abdominal pain were the commonest symptoms. Six patients had a positive family history, while four others were diagnosed as index cases of familial adenomatous polyposis. Rectal tumours made up 43% of all colorectal cancers diagnosed. Seventy percent of patients presented at an advanced stage, but curative resection was attempted for 29 patients. Eight underwent palliative resections, 1 had an ileostomy while another underwent a bypass procedure. Eleven patients have died, 14 had no evidence of recurrent disease while 3 were still alive with recurrent disease. CONCLUSION: Age does not affect survival, and early endoscopy is recommended for all with persistent symptoms. Early diagnosis, radical resection and adjuvant therapy still form the cornerstone in management of colorectal cancer in this age group. PMID- 15109386 TI - Small bowel malignancies: a review of 29 patients at a single centre. AB - OBJECTIVE: We analysed the incidence, clinical presentation and outcome of small bowel malignancies treated in the department. PATIENTS AND METHODS: From July 1993 to December 2001, 29 patients with small bowel malignancies were operated upon. The median age was 60 years (30-87 years), and there were 11 males and 18 females. The more frequently used diagnostic investigations included barium contrast studies, and CT scanning. RESULTS: The most frequent complaint was abdominal pain, though other symptoms like abdominal mass, weight loss and change in bowel habit may also be present. Intestinal obstruction necessitated emergency operation in some cases. One case was treated as Crohn's stricture and two others, as irritable bowel syndrome for nearly 6 months before referral was made for surgery. Fifteen cases were secondaries to the small bowel, and fourteen were primary small bowel malignancies. The secondaries were metastases from colorectal cancers (8), gynaecological cancers (5), breast (1) and transitional cell carcinomas (1). Of the 14 primary small bowel malignancies, there were 8 lymphomas, 4 adenocarcinomas and 2 neuroendocrine tumours. Eleven of these patients were alive at mean follow-up of 25 months (1-97 months) and 2 were lost to follow-up. Only one of the secondary small bowel malignancies was alive at the time of review. CONCLUSION: Small bowel malignancy is a rate entity with pre operative diagnosis remaining difficult. Early surgical intervention with a high index of suspicion is required to improve survival. PMID- 15109387 TI - Long-term urinary dysfunction after rectal cancer surgery. AB - OBJECTIVE: Urinary dysfunction is a well-known complication of rectal surgery, secondary to injury to the autonomic nervous plexus. The object of this study was to evaluate the incidence, prevalence and type of micturition disorders following rectal cancer surgery and their reversibility during long-term follow-up. PATIENTS AND METHODS: A prospective study of 45 patients who underwent surgery for rectal cancer between 1993 and 1998 was undertaken. Those with pre-operative urinary dysfunction were excluded after sequential uroflowmetry and clinical interview. Ten of the surgical interventions were high anterior resections, 18 low anterior resections, and 17 abdominoperineal amputations. Pre-operative radiotherapy was performed in 47.9% of patients. All patients underwent sequential uroflowmetry and a clinical interview 3 and 12 months after the intervention. Subjects who presented micturition disorders underwent urodynamic examination The follow up period was three years. RESULTS: Three months after surgery alterations were found in 14 (31.3%) patients; the most frequent were stress incontinence, urinary tenesmus and the urge to urinate. At the 12-month assessment only 6 (13.3%) patients had urinary symptomatology or uroflowmetry abnormalities. After three years, micturition disorders persisted in 3 (6.6%) patients. CONCLUSION: Urinary dysfunction after rectal cancer excision is associated with a high degree of reversibility. Seventy-eight percent of the alterations detected after three months and 50% of those that persisted after a year disappeared during follow up. PMID- 15109388 TI - Parastomal hernia repair. AB - OBJECTIVE: The aim of this study was to audit the results of parastomal hernia repair. METHODS: A retrospective review was made of all patients having parastomal hernia repair at 2 teaching hospitals over a 12-year period. RESULTS: Forty-three patients had 51 parastomal hernia repairs. Three types of hernia repair were used: 14 (28%) local suture, 19 (37%) local mesh repair and 18 (35%) stoma relocation. Three patients died and significant complications occurred in 65% of repairs. Parastomal hernia recurred in 18 (38%) of repairs. This was greatest with a sutured repair (59%) and least in patients having stoma relocation (24%). Repair with mesh was associated with a 39% recurrence rate. No significant statistical difference was found for the results of these repairs. CONCLUSION: Parastomal hernia repair is associated with high rates of morbidity, mortality and recurrence. Repair should not be undertaken without a good indication. PMID- 15109389 TI - Extramammary Paget's disease of the perianal region. AB - OBJECTIVE: Perianal Paget's disease (PPD) is a rare entity. The standard treatment for either in situ or invasive extra mammary Paget's disease (EMPD) is surgical excision. Local recurrence and morbidity from surgery, especially in the elderly, can, however, be high. The aim of this article is to review our experience with PPD and question the currently preferred treatment approaches in light of its histopathology and therapeutic outcome. PATIENTS AND METHODS: A chart review of our patients with PPD from 1996 to 2002 was carried out to determine their outcome after treatment. Data from review of the literature are presented. RESULTS: Five patients with in situ disease (four females, median age 68 years) were diagnosed as having PPD. A complete surgical excision was attempted in 4 patients and the fifth was treated by photodynamic therapy. At present, all patients are alive, two are free of disease, one has persistent disease and two have local recurrence. CONCLUSION: Considering the significant rate of recurrence even after wide local excision, the extent of surgery needed and the good prognosis with long-term survival, we question whether nonsurgical modalities should be considered in place of surgery as primary treatment for noninvasive PPD, with radical surgery being reserved for failures or invasive disease. PMID- 15109390 TI - Functional outcome following lateral internal anal sphincterotomy for chronic anal fissure. PMID- 15109391 TI - Stapled anopexy for short segment Hirschprungs disease in adults. AB - Traditional treatment of short segment Hirschprungs disease in adult consists of major resectional procedures, often with the use of a temporary stoma. Patients with this disease may have significant morbidities that increase their risk of post-operative morbidity and mortality. In an attempt to minimize the procedural related morbidity, we describe the application of the stapled anopexy technique to treat short segment Hirschprungs disease. PMID- 15109393 TI - Rapid prefractionation of complex protein lysates with centrifugal membrane adsorber units improves the resolving power of 2D-PAGE-based proteome analysis. AB - BACKGROUND: Two-dimensional gel electrophoresis (2D-PAGE) has proven over the years to be a reliable and efficient method for separation of hundreds of proteins based on charge and mass. Nevertheless, the complexity of even the simplest proteomes limits the resolving power of 2D-PAGE. This limitation can be partially alleviated by sample prefractionation using a variety of techniques. RESULTS: Here, we have used Vivapure Ion Exchange centrifugal adsorber units to rapidly prefractionate total fission yeast protein lysate based on protein charge. Three fractions were prepared by stepwise elution with increasing sodium chloride concentrations. Each of the fractions, as well as the total lysate, were analyzed by 2D-PAGE. This simple prefractionation procedure considerably increased the resolving power of 2D-PAGE. Whereas 308 spots could be detected by analysing total protein lysate, 910 spots were observed upon prefractionation. Thorough gel image analysis demonstrated that prefractionation visualizes an additional set of 458 unique fission yeast proteins not detected in whole cell lysate. CONCLUSIONS: Prefractionation with Vivapure Q spin columns proved to be a simple, fast, reproducible, and cost-effective means of increasing the resolving power of 2D-PAGE using standard laboratory equipment. PMID- 15109394 TI - Recombinant adeno-associated virus type 2-mediated gene delivery into the Rpe65-/ knockout mouse eye results in limited rescue. AB - BACKGROUND: Leber's congenital amaurosis (LCA) is a severe form of retinal dystrophy. Mutations in the RPE65 gene, which is abundantly expressed in retinal pigment epithelial (RPE) cells, account for approximately 10-15% of LCA cases. In this study we used the high turnover, and rapid breeding and maturation time of the Rpe65-/- knockout mice to assess the efficacy of using rAAV-mediated gene therapy to replace the disrupted RPE65 gene. The potential for rAAV-mediated gene treatment of LCA was then analyzed by determining the pattern of RPE65 expression, the physiological and histological effects that it produced, and any improvement in visual function. METHODS: rAAV.RPE65 was injected into the subretinal space of Rpe65-/- knockout mice and control mice. Histological and immunohistological analyses were performed to evaluate any rescue of photoreceptors and to determine longevity and pattern of transgene expression. Electron microscopy was used to examine ultrastructural changes, and electroretinography was used to measure changes in visual function following rAAV.RPE65 injection. RESULTS: rAAV-mediated RPE65 expression was detected for up to 18 months post injection. The delivery of rAAV.RPE65 to Rpe65-/- mouse retinas resulted in a transient improvement in the maximum b-wave amplitude under both scotopic and photopic conditions (76% and 59% increase above uninjected controls, respectively) but no changes were observed in a-wave amplitude. However, this increase in b-wave amplitude was not accompanied by any slow down in photoreceptor degeneration or apoptotic cell death. Delivery of rAAV.RPE65 also resulted in a decrease in retinyl ester lipid droplets and an increase in short wavelength cone opsin-positive cells, suggesting that the recovery of RPE65 expression has long-term benefits for retinal health. CONCLUSION: This work demonstrated the potential benefits of using the Rpe65-/- mice to study the effects and mechanism of rAAV.RPE65-mediated gene delivery into the retina. Although the functional recovery in this model was not as robust as in the dog model, these experiments provided important clues about the long-term physiological benefits of restoration of RPE65 expression in the retina. PMID- 15109395 TI - L1 and HERV-W retrotransposons are hypomethylated in human ovarian carcinomas. AB - Wide-spread hypomethylation of CpG dinucleotides is characteristic of many cancers. Retrotransposons have been identified as potential targets of hypomethylation during cellular transformation. We report the results of an preliminary examination of the methylation status of CpG dinucleotides associated with the L1 and HERV-W retrotransposons in benign and malignant human ovarian tumors. We find a reduction in the methylation of CpG dinucleotides within the promoter regions of these retroelements in malignant relative to non-malignant ovarian tissues. Consistent with these results, we find that relative L1 and HERV W expression levels are elevated in representative samples of malignant vs. non malignant ovarian tissues. PMID- 15109396 TI - Comparison of gene expression in HCT116 treatment derivatives generated by two different 5-fluorouracil exposure protocols. AB - BACKGROUND: Established colorectal cancer cell lines subjected to different 5 fluorouracil (5-FU) treatment protocols are often used as in vitro model systems for investigations of downstream cellular responses to 5-FU and to generate 5-FU resistant derivatives for the investigation of biological mechanisms involved in drug resistance. We subjected HCT116 colon cancer cells to two different 5-FU treatment protocols in an attempt to generate resistant derivatives: one that simulated the clinical bolus regimens using clinically-achievable 5-FU levels, the other that utilized serial passage in the presence of increasing 5-FU concentrations (continuous exposure). HCT116 Bolus3, ContinB, and ContinD, corresponding to independently-derived cell lines generated either by bolus exposure or continuous exposure, respectively, were characterized for growth- and apoptosis-associated phenotypes, and gene expression using 8.5 K oligonucleotide microarrays. Comparative gene expression analyses were done in order to determine if transcriptional profiles for the respective treatment derivatives were similar or substantially different, and to identify the signaling and regulatory pathways involved in mediating the downstream response to 5-FU exposure and possibly involved in development of resistance. RESULTS: HCT116 ContinB and ContinD cells were respectively 27-fold and >100-fold more resistant to 5-FU and had reduced apoptotic fractions in response to transient 5-FU challenge compared to the parental cell line, whereas HCT116 Bolus3 cells were not resistant to 5-FU after 3 cycles of bolus 5-FU treatment and had the same apoptotic response to transient 5-FU challenge as the parental cell line. However, gene expression levels and expression level changes for all detected genes in Bolus3 cells were similar to those seen in both the ContinB (strongest correlation) and ContinD derivatives, as demonstrated by correlation and cluster analyses. Regulatory pathways having to do with 5-FU metabolism, apoptosis, and DNA repair were among those that were affected by 5-FU treatment. CONCLUSION: All HCT116 derivative cell lines demonstrated similar transcriptional profiles, despite the facts that they were generated by two different 5-FU exposure protocols and that the bolus exposure derivative had not become resistant to 5-FU. Selection pressures on HCT116 cells as a result of 5-FU challenge are thus similar for both treatment protocols. PMID- 15109397 TI - Costs and risk factors for ventilator-associated pneumonia in a Turkish university hospital's intensive care unit: a case-control study. AB - BACKGROUND: Ventilator-associated pneumonia (VAP) which is an important part of all nosocomial infections in intensive care unit (ICU) is a serious illness with substantial morbidity and mortality, and increases costs of hospital care. We aimed to evaluate costs and risk factors for VAP in adult ICU. METHODS: This is a three year retrospective case-control study. The data were collected between 01 January 2000 and 31 December 2002. During the study period, 132 patients were diagnosed as nosocomial pneumonia of 731 adult medical-surgical ICU patients. Of these only 37 VAP patients were assessed, and multiple nosocomially infected patients were excluded from the study. Sixty non-infected ICU patients were chosen as control patients. RESULTS: Median length of stay in ICU in patients with VAP and without were 8.0 (IQR: 6.5) and 2.5 (IQR: 2.0) days respectively (P < 0.0001). Respiratory failure (OR, 11.8; 95%, CI, 2.2-62.5; P < 0.004), coma in admission (Glasgow coma scale < 9) (OR, 17.2; 95% CI, 2.7-107.7; P < 0.002), depressed consciousness (OR, 8.8; 95% CI, 2.9-62.5; P < 0.02), enteral feeding (OR, 5.3; 95% CI, 1.0-27.3; P = 0.044) and length of stay (OR, 1.3; 95% CI, 1.0 1.7; P < 0.04) were found as important risk factors. Most commonly isolated microorganism was methicillin resistant Staphylococcus aureus (30.4%). Mortality rates were higher in patients with VAP (70.3%) than the control patients (35.5%) (P < 0.003). Mean cost of patients with and without VAP were 2832.2+/-1329.0 and 868.5+/-428.0 US Dollars respectively (P < 0.0001). CONCLUSION: Respiratory failure, coma, depressed consciousness, enteral feeding and length of stay are independent risk factors for developing VAP. The cost of VAP is approximately five-fold higher than non-infected patients. PMID- 15109398 TI - Survey on schizophrenia treatment in Mexico: perception and antipsychotic prescription patterns. AB - BACKGROUND: Since the introduction of antipsychotics, especially the so called atypicals, the treatment of schizophrenia has shown important improvements. At the present time, it is preferred to label clozapine and other antipsychotics sharing similar profiles as second-generation antipsychotics (SGAs). These medications have been proposed by some experts as a first line treatment for schizophrenia. It is critical to have reliable data about antipsychotic prescription in Mexico and to create management guidelines based on expert meetings and not only on studies carried out by the pharmaceutical industry. Only this approach will help to make the right decisions for the treatment of schizophrenia. METHODS: A translated version of Rabinowitz's survey was used to evaluate antipsychotic prescription preferences and patterns in Mexican psychiatrists. The survey questionnaire was sent by mail to 200 psychiatrists from public institutions and private practice in Mexico City and Guadalajara, Mexico. RESULTS: Recommendations for antipsychotics daily doses at different stages of the treatment of schizophrenia varied widely. Haloperidol was considered as the first choice for the treatment of positive symptoms. On the contrary, risperidone was the first option for negative symptoms. For a patient with a high susceptibility for developing extrapyramidal symptoms (EPS), risperidone was the first choice. It was also considered that SGAs had advantages over typical antipsychotics in the management of negative symptoms, cognitive impairment and fewer EPS.Besides, there was a clear tendency for prescribing typical antipsychotics at higher doses than recommended and inadequate doses for the atypical ones. CONCLUSIONS: Some of the obstacles for the prescription of SGAs include their high cost, deficient knowledge about their indications and dosage, the perception of their being less efficient for the treatment of positive symptoms and the resistance of some Mexican physicians to change their prescription pattern. It is necessary to reach a consensus, in order to establish and standardize the treatment of schizophrenia, based on the information reported in clinical trials and prevailing economic conditions in Mexico. PMID- 15109399 TI - Low dose intravenous minocycline is neuroprotective after middle cerebral artery occlusion-reperfusion in rats. AB - BACKGROUND: Minocycline, a semi-synthetic tetracycline antibiotic, is an effective neuroprotective agent in animal models of cerebral ischemia when given in high doses intraperitoneally. The aim of this study was to determine if minocycline was effective at reducing infarct size in a Temporary Middle Cerebral Artery Occlusion model (TMCAO) when given at lower intravenous (IV) doses that correspond to human clinical exposure regimens. METHODS: Rats underwent 90 minutes of TMCAO. Minocycline or saline placebo was administered IV starting at 4, 5, or 6 hours post TMCAO. Infarct volume and neurofunctional tests were carried out at 24 hr after TMCAO using 2,3,5-triphenyltetrazolium chloride (TTC) brain staining and Neurological Score evaluation. Pharmacokinetic studies and hemodynamic monitoring were performed on minocycline-treated rats. RESULTS: Minocycline at doses of 3 mg/kg and 10 mg/kg IV was effective at reducing infarct size when administered at 4 hours post TMCAO. At doses of 3 mg/kg, minocycline reduced infarct size by 42% while 10 mg/kg reduced infarct size by 56%. Minocycline at a dose of 10 mg/kg significantly reduced infarct size at 5 hours by 40% and the 3 mg/kg dose significantly reduced infarct size by 34%. With a 6 hour time window there was a non-significant trend in infarct reduction. There was a significant difference in neurological scores favoring minocycline in both the 3 mg/kg and 10 mg/kg doses at 4 hours and at the 10 mg/kg dose at 5 hours. Minocycline did not significantly affect hemodynamic and physiological variables. A 3 mg/kg IV dose of minocycline resulted in serum levels similar to that achieved in humans after a standard 200 mg dose. CONCLUSIONS: The neuroprotective action of minocycline at clinically suitable dosing regimens and at a therapeutic time window of at least 4-5 hours merits consideration of phase I trials in humans in view of developing this drug for treatment of stroke. PMID- 15109400 TI - Coronary age as a risk factor in the modified Framingham risk score. AB - BACKGROUND: Clinical guidelines emphasize risk assessment as vital to patient selection for medical primary intervention. However, risk assessment methods are restricted in their ability to predict further coronary events. The most widely accepted tool in the United States is the Framingham risk score. In these equations age is a powerful risk factor. Although the extent of coronary atherosclerosis increases with age, there is large inter-individual variability in the rate of development and progression of this disease. This fact limits the utility of Framingham scoring when applied to individuals. Electron beam tomography (EBT), which measures coronary calcium, provides a non-invasive method for assessing coronary plaque burden, thus offering the possibility of providing a more accurate estimate of an individual's "arterial age" than from chronological age alone. METHODS: In this paper we discuss a new and simple method for incorporating the coronary calcium score (CCS) to modify the Framingham Risk Assessment (FRA). Using this method, a coronary artery calcium (CAC) age equivalent is generated that replaces chronological age in Framingham scoring. RESULTS AND DISCUSSION: Using a percentile table of CCS scores by age group and sex, individuals are matched to the age group whose calcium score most closely approximates their own individual score. The original 10-year absolute risk score of a 65-year old man with a CCS of 6 based on chronological age is 10%, whereas the modified absolute risk score based on CAC age equivalents is 2%. CONCLUSION: Our approach of replacing chronological age with CAC age equivalents in the Framingham equations possesses simplicity of application combined with precision. Physicians can easily derive adjusted Framingham risk scores and prescribe intervention methods based on patients' ten-year risks. The adjusted ten-year risks are likely to be more accurate than unadjusted risks since they are based on coronary calcium score information. The modified FRA approach not only may increase the predicted risk for some patients, but also may decrease the predicted risk for others, making it a more precise adjustment than other methods. PMID- 15109401 TI - Up-regulation of the chemokine CCL21 in the skin of subjects exposed to irritants. AB - BACKGROUND: Expression of murine CCL21 by dermal lymphatic endothelial cells (LEC) has been demonstrated to be one of the most important steps in Langerhans cell emigration from skin. Previously, our group and others have found that this chemokine is up-regulated in different human inflammatory skin diseases mediated by diverse specific immune responses. This study was carried out to investigate the involvement of CCL21 in human skin after challenge with irritant agents responsible for inducing Irritant Contact Dermatitis (ICD). RESULTS: Eleven normal individuals were challenged with different chemical or physical irritants. Two patients with Allergic Contact Dermatitis (ACD) were also challenged with the relevant antigen in order to have a positive control for CCL21 expression. Macroscopic as well as microscopic responses were evaluated. We observed typical ICD responses with mostly mononuclear cells in perivascular areas, but a predominance of polymorphonuclear cells away from the inflamed blood vessels and in the epidermis at 24 hours. Immunohistochemical studies showed up-regulation of CCL21 by lymphatic endothelial cells in all the biopsies taken from ICD and ACD lesions compared to normal skin. Kinetic study at 10, 48, 96 and 168 hours after contact with a classical irritant (sodium lauryl sulphate) showed that the expression of CCL21 was increased in lymphatic vessels at 10 hours, peaked at 48 hours, and then gradually declined. There was a strong correlation between CCL21 expression and the macroscopic response (r = 0.69; p = 0.0008), but not between CCL21 and the number of infiltrating cells in the lesions. CONCLUSIONS: These results provide new evidence for the role of CCL21 in inflammatory processes. Since the up-regulation of this chemokine was observed in ICD and ACD, it is tempting to speculate that this mechanism operates independently of the type of dermal insult, facilitating the emigration of CCR7+ cells. PMID- 15109402 TI - Clinical trials and novel pathogens: lessons learned from SARS. AB - During the recent global outbreak of severe acute respiratory syndrome (SARS), thousands of patients received treatments of uncertain efficacy and known toxicity such as ribavirin and corticosteroids. Despite this, no controlled clinical trials assessing the efficacy of these agents were conducted. If a second global SARS outbreak occurred, clinicians would not have controlled data on which to base therapeutic decisions. We discuss the unique methodologic and logistical challenges faced by researchers who attempt to conduct controlled trials of therapeutic agents during an outbreak of a novel or unknown infectious pathogen. We draw upon our own experience in attempting to conduct a randomized controlled trial (trial) of ribavirin therapy for SARS and discuss the lessons learned. Strategies to facilitate future clinical trials during outbreaks of unknown or novel pathogens are also presented. PMID- 15109403 TI - SARS transmission and hospital containment. AB - An outbreak of severe acute respiratory syndrome (SARS) was detected in Singapore at the beginning of March 2003. The outbreak, initiated by a traveler to Hong Kong in late February 2003, led to sequential spread of SARS to three major acute care hospitals in Singapore. The critical factor in containing this outbreak was early detection and complete assessment of movements and follow-up of patients, healthcare workers, and visitors who were contacts. Visitor records were important in helping identify exposed persons who could carry the infection into the community. In the three hospital outbreaks, three different containment strategies were used to contain spread of infection: closing an entire hospital, removing all potentially infected persons to a dedicated SARS hospital, and managing exposed persons in place. On the basis of this experience, if a nosocomial outbreak is detected late, a hospital may need to be closed in order to contain spread of the disease. Outbreaks detected early can be managed by either removing all exposed persons to a designated location or isolating and managing them in place. PMID- 15109404 TI - The RUsick2 Foodborne Disease Forum for syndromic surveillance. AB - The RUsick2 Foodborne Disease Forum at the National Food Safety and Toxicology Center increased reporting of foodborne diseases to more than four times the rate seen in the previous 2 years. Since November 2002, the Forum has allowed pilot area residents with sudden-onset vomiting or diarrhea to share and compare information regarding what they ate and did before becoming sick. The purpose is to identify a common food source, perhaps resulting in identifying a cluster of persons who ate the same contaminated food item. Such information can assist health departments in detecting foodborne outbreaks while the possibility for intervention remains. PMID- 15109405 TI - Reemerging leptospirosis, California. AB - Leptospirosis is a reemerging infectious disease in California. Leptospirosis is the most widespread zoonosis throughout the world, though it is infrequently diagnosed in the continental United States. From 1982 to 2001, most reported California cases occurred in previously healthy young adult white men after recreational exposures to contaminated freshwater. We report five recent cases of human leptospirosis acquired in California, including the first documented common source outbreak of human leptospirosis acquired in this state, and describe the subsequent environmental investigation. Salient features in the California cases include high fever with uniform renal impairment and mild hepatitis. Because leptospirosis can progress rapidly if untreated, this reemerging infection deserves consideration in febrile patients with a history of recreational freshwater exposure, even in states with a low reported incidence of infection. PMID- 15109406 TI - Coronaviridae and SARS-associated coronavirus strain HSR1. AB - During the recent severe acute respiratory (SARS) outbreak, the etiologic agent was identified as a new coronavirus (CoV). We have isolated a SARS-associated CoV (SARS-CoV) strain by injecting Vero cells with a sputum specimen from an Italian patient affected by a severe pneumonia; the patient traveled from Vietnam to Italy in March 2003. Ultrastructural analysis of infected Vero cells showed the virions within cell vesicles and around the cell membrane. The full-length viral genome sequence was similar to those derived from the Hong-Kong Hotel M isolate. By using both real-time reverse transcription-polymerase chain reaction TaqMan assay and an infectivity plaque assay, we determined that approximately 360 viral genomes were required to generate a PFU. In addition, heparin (100 microg/mL) inhibited infection of Vero cells by 50%. Overall, the molecular and biologic characteristics of the strain HSR1 provide evidence that SARS-CoV forms a fourth genetic coronavirus group with distinct genomic and biologic features. PMID- 15109407 TI - Laboratory analysis of tularemia in wild-trapped, commercially traded prairie dogs, Texas, 2002. AB - Oropharyngeal tularemia was identified as the cause of a die-off in captured wild prairie dogs at a commercial exotic animal facility in Texas. From this point source, Francisella tularensis-infected prairie dogs were traced to animals distributed to the Czech Republic and to a Texas pet shop. F. tularensis culture isolates were recovered tissue specimens from 63 prairie dogs, including one each from the secondary distribution sites. Molecular and biochemical subtyping indicated that all isolates were F. tularensis subsp. holarctica (Type B). Microagglutination assays detected antibodies against F. tularensis, with titers as great as 1:4,096 in some live animals. All seropositive animals remained culture positive, suggesting that prairie dogs may act as chronic carriers of F. tularensis. These findings demonstrate the need for additional studies of tularemia in prairie dogs, given the seriousness of the resulting disease, the fact that prairie dogs are sold commercially as pets, and the risk for pet-to human transmission. PMID- 15109408 TI - Monkeypox transmission and pathogenesis in prairie dogs. AB - During May and June 2003, the first cluster of human monkeypox cases in the United States was reported. Most patients with this febrile vesicular rash illness presumably acquired the infection from prairie dogs. Monkeypox virus was demonstrated by using polymerase chain reaction in two prairie dogs in which pathologic studies showed necrotizing bronchopneumonia, conjunctivitis, and tongue ulceration. Immunohistochemical assays for orthopoxviruses demonstrated abundant viral antigens in surface epithelial cells of lesions in conjunctiva and tongue, with less amounts in adjacent macrophages, fibroblasts, and connective tissues. Viral antigens in the lung were abundant in bronchial epithelial cells, macrophages, and fibroblasts. Virus isolation and electron microscopy demonstrated active viral replication in lungs and tongue. These findings indicate that both respiratory and direct mucocutaneous exposures are potentially important routes of transmission of monkeypox virus between rodents and to humans. Prairie dogs offer insights into transmission, pathogenesis, and new vaccine and treatment trials because they are susceptible to severe monkeypox infection. PMID- 15109409 TI - Acute spotted fever rickettsiosis among febrile patients, Cameroon. AB - Although potential arthropod vectors are abundant in Cameroon, acute febrile illnesses are rarely evaluated for arboviral or rickettsial infections. Serum samples from 234 acutely febrile patients at clinics in Tiko and Buea, Cameroon, were examined for antibodies to Rickettsia africae and African alphaviruses and flaviviruses. These serum samples did not contain antibodies against typhoid, and blood malarial parasites were not detected. Serum samples of 32% contained immunoglobulin M antibodies reactive with R. africae by immunofluorescence assay and were reactive with outer membrane proteins A and B of R. africae by immunoblotting. These findings established a diagnosis of acute rickettsiosis, most likely African tick-bite fever. Hemagglutination inhibition testing of the serum samples also detected antibodies to Chikungunya virus (47%) and flaviviruses (47%). High prevalence of antibodies to arboviruses may represent a major, previously unrecognized public health problem in an area where endemic malaria and typhoid fever have been the principal diagnostic considerations. PMID- 15109410 TI - Genomic changes of Chagas disease vector, South America. AB - We analyzed the main karyologic changes that have occurred during the dispersion of Triatoma infestans, the main vector of Chagas disease. We identified two allopatric groups, named Andean and non-Andean. The Andean specimens present C heterochromatic blocks in most of their 22 chromosomes, whereas non-Andean specimens have only 4-7 autosomes with C-banding. These heterochromatin differences are the likely cause of a striking DNA content variation (approximately 30%) between Andean and non-Andean insects. Our study, together with previous historical and genetic data, suggests that T. infestans was originally a sylvatic species, with large quantities of DNA and heterochromatin, inhabiting the Andean region of Bolivia. However, the spread of domestic T. infestans throughout the non-Andean regions only involved insects with an important reduction of heterochromatin and DNA amounts. We propose that heterochromatin and DNA variation mainly reflected adaptive genomic changes that contribute to the ability of T. infestans to survive, reproduce, and disperse in different environments. PMID- 15109411 TI - Internet use and epidemiologic investigation of gastroenteritis outbreak. AB - In March 2000, a large outbreak of gastroenteritis occurred in a community where a regional computer network provides free Internet access for 42% of the households. We conducted an epidemiologic investigation using the Internet for data collection. Norovirus was identified in stool samples of nine patients but not in the municipal water supply. Of households with access to the network, 19% participated in the survey. The overall attack rate by household was 63%. Drinking water from the nonchlorinated community water system was associated with illness (relative risk [RR] 1.6; 95% confidence interval [CI] 1.1 to 2.2); drinking water only from a private well was associated with decreased likelihood of illness (RR 0.3; 95% CI 0.1 to 0.8). Data collection through the Internet was efficient. Internet surveys may become more common in epidemiologic investigations and have the potential to provide data rapidly, enabling appropriate public health action. However, methods should be developed to increase response rates and minimize bias. PMID- 15109412 TI - Correlating epidemiologic trends with the genotypes causing meningococcal disease, Maryland. AB - Epidemic meningococcal infection is generally caused by single clones; whether nonepidemic increases in infection are clonal is unknown. We studied the molecular epidemiology of meningococcal infection during a period that the incidence increased in two age groups. Serogroup C and Y meningococcal isolates were analyzed by pulsed-field gel electrophoresis and multilocus sequence typing. From 1992 to 1999, 96.4% (27/28) of serogroup C isolates from persons 15-24 years of age were in clonal group 1, compared with 65.6% (21/32) of isolates from persons < or =14 years, and 64.3% (9/14) of isolates from adults > or =25 years (p < or = 0.01). The proportion of clonal group 2 serogroup Y strains increased from 7.7% (1/13) in 1992 to 1993 to 52.0% (13/25) in 1998 to 1999 (p < 0.01). The nonepidemic age-specific increases in serogroup C meningococcal infection in Maryland were clonal in nature and the changes in serogroup Y incidence were associated with a shift in the genotypes of strains causing invasive disease. PMID- 15109413 TI - Legionella infection risk from domestic hot water. AB - We investigated Legionella and Pseudomonas contamination of hot water in a cross sectional multicentric survey in Italy. Chemical parameters (hardness, free chlorine, and trace elements) were determined. Legionella spp. were detected in 33 (22.6%) and Pseudomonas spp. in 56 (38.4%) of 146 samples. Some factors associated with Legionella contamination were heater type, tank distance and capacity, water plant age, and mineral content. Pseudomonas presence was influenced by water source, hardness, free chlorine, and temperature. Legionella contamination was associated with a centralized heater, distance from the heater point >10 m, and a water plant >10 years old. Furthermore, zinc levels of <100 microg/L and copper levels of >50 microg/L appeared to be protective against Legionella colonization [corrected]. Legionella species and serogroups were differently distributed according to heater type, water temperature, and free chlorine, suggesting that Legionella strains may have a different sensibility and resistance to environmental factors and different ecologic niches. PMID- 15109414 TI - Cysticercosis-related deaths, California. AB - Cysticercosis is an increasingly important disease in the United States, but information on the occurrence of related deaths is limited. We examined data from California death certificates for the 12-year period 1989-2000. A total of 124 cysticercosis deaths were identified, representing a crude 12-year death rate of 3.9 per million population (95% confidence interval [CI] 3.2 to 4.6). Eighty-two (66%) of the case-patients were male; 42 (34%) were female. The median age at death was 34.5 years (range 7-81 years). Most patients (107, 86.3%) were foreign born, and 90 (72.6%) had emigrated from Mexico. Seventeen (13.7%) deaths occurred in U.S.-born residents. Cysticercosis death rates were higher in Latino residents of California (13.0/106) than in other racial/ethnic groups (0.4/106), in males (5.2/106) than in females (2.7/106), and in persons >14 years of age (5.0/106). Cysticercosis is a preventable cause of premature death, particularly among young Latino persons in California and may be a more common cause of death in the United States than previously recognized. PMID- 15109415 TI - Amoebae-resisting bacteria isolated from human nasal swabs by amoebal coculture. AB - Amoebae feed on bacteria, and few bacteria can resist their microbicidal ability. Amoebal coculture could therefore be used to selectively grow these amoebae resisting bacteria (ARB), which may be human pathogens. To isolate new ARB, we performed amoebal coculture from 444 nasal samples. We recovered 7 (1.6%) ARB from 444 nasal swabs, including 4 new species provisionally named Candidatus Roseomonas massiliae, C. Rhizobium massiliae, C. Chryseobacterium massiliae, and C. Amoebinatus massiliae. The remaining isolates were closely related to Methylobacterium extorquens, Bosea vestrii, and Achromobacter xylosoxidans. Thus, amoebal coculture allows the recovery of new bacterial species from heavily contaminated samples and might be a valuable approach for the recovery of as-yet unrecognized emerging pathogens from clinical specimens. PMID- 15109416 TI - Neutralizing antibodies and Sin Nombre virus RNA after recovery from hantavirus cardiopulmonary syndrome. AB - Patients who later have a mild course of hantavirus cardiopulmonary syndrome (HCPS) are more likely to exhibit a high titer of neutralizing antibodies against Sin Nombre virus (SNV), the etiologic agent of HCPS, at the time of hospital admission. Because administering plasma from patients who have recovered from HCPS to those in the early stages of disease may be an advantageous form of passive immunotherapy, we examined the neutralizing antibody titers of 21 patients who had recovered from SNV infection. Even 1,000 days after admission to the hospital, 6 of 10 patients had titers of 800 or higher, with one sample retaining a titer of 3,200 after more than 1,400 days. None of the convalescent phase serum samples contained detectable viral RNA. These results confirm that patients retain high titers of neutralizing antibodies long after recovery from SNV infection. PMID- 15109417 TI - First reported prairie dog-to-human tularemia transmission, Texas, 2002. AB - A tularemia outbreak, caused by Francisella tularensis type B, occurred among wild-caught, commercially traded prairie dogs. F. tularensis microagglutination titers in one exposed person indicated recent infection. These findings represent the first evidence for prairie-dog-to-human tularemia transmission and demonstrate potential human health risks of the exotic pet trade. PMID- 15109418 TI - Predicting quarantine failure rates. AB - Preemptive quarantine through contact-tracing effectively controls emerging infectious diseases. Occasionally this quarantine fails, however, and infected persons are released. The probability of quarantine failure is typically estimated from disease-specific data. Here a simple, exact estimate of the failure rate is derived that does not depend on disease-specific parameters. This estimate is universally applicable to all infectious diseases. PMID- 15109419 TI - Patient data, early SARS epidemic, Taiwan. AB - Of the first 10 patients in the epidemic of severe acute respiratory syndrome (SARS) in Taiwan, 4 were closely associated with a SARS patient in an airplane. Loose stools or diarrhea, hemophagocytosis syndrome, and high serum levels of interleukin (IL)-6, IL-8, and tumor necrosis factor-a associated with lung lesions were found in all 10 patients. PMID- 15109420 TI - B-virus and free-ranging macaques, Puerto Rico. AB - In Puerto Rico, risk for transmission of B-virus from free-ranging rhesus monkeys to humans has become a serious challenge. An incident with an injured rhesus monkey, seropositive for B-virus, resulted in inappropriate administration of antiviral postexposure prophylaxis. This incident underscores the importance of education about risks associated with interactions between humans and nonhuman primates. PMID- 15109421 TI - Human metapneumovirus-associated atypical pneumonia and SARS. AB - Acute pneumonia developed in a previously healthy man during the outbreak of severe acute respiratory syndrome (SARS) in southern China in March 2003. Antibiotic treatment was ineffective, and he died 8 days after illness onset. Human metapneumovirus was isolated from lung tissue. No other pathogen was found. Other etiologic agents should thus be sought in apparent SARS cases when coronavirus infection cannot be confirmed. PMID- 15109422 TI - Q fever endocarditis in HIV-infected patient. AB - We describe a case of Q fever endocarditis in an HIV-infected patient. The case was treated successfully with valvular replacement and a combination of doxycycline and hydroxychloroquine. We review the current literature on Q fever endocarditis, with an emphasis on the co-infection of HIV and Coxiella burnetii. PMID- 15109423 TI - Bovine necrotic vulvovaginitis associated with Porphyromonas levii. AB - An outbreak of bovine necrotic vulvovaginitis associated with Porphyromonas levii, an emerging animal and human pathogen, affected 32 cows on a dairy farm in the northeast of Israel. Five animals had to be culled. This report appears to be the first that associates P. levii with bovine necrotic vulvovagnitis. PMID- 15109424 TI - Neurocysticercosis in Oregon, 1995-2000. AB - The unexpected death of a teenager from neurocysticercosis prompted an investigation of this disease in Oregon. We found 89 hospitalizations, 43 newly diagnosed cases, and 6 deaths from 1995 to 2000. At least five cases occurred in persons who had not traveled or lived outside the United States. Enhanced surveillance for neurocysticercosis is warranted. PMID- 15109425 TI - Imported cutaneous diphtheria, United Kingdom. AB - Cutaneous diphtheria is endemic in tropical countries but unusual in the United Kingdom. Four cases occurred in the United Kingdom within 2 months in 2002. Because cutaneous diphtheria causes outbreaks of both cutaneous and pharyngeal forms, early diagnosis is essential for implementing control measures; high diphtheria vaccination coverage must also be maintained. PMID- 15109426 TI - Antibiotic selection pressure and resistance in Streptococcus pneumoniae and Streptococcus pyogenes. AB - We correlated outpatient antibiotic use with prevalence of penicillin nonsusceptible Streptococcus pneumoniae (PNSP), macrolide-resistant S. pneumoniae (MRSP), and macrolide-resistant S. pyogenes (MRGAS) in 20 countries. Total antibiotic use was correlated with PNSP (r = 0.75; p < 0.001), as was macrolide use with MRSP (r = 0.88; p < 0.001) and MRGAS (r = 0.71; p = 0.004). Streptococcal resistance is directly associated with antibiotic selection pressure on a national level. PMID- 15109427 TI - Enterotoxin-producing Escherichia coli O169:H41, United States. AB - From 1996 to 2003, 16 outbreaks of enterotoxigenic Escherichia coli (ETEC) infections in the United States and on cruise ships were confirmed. E. coli serotype O169:H41 was identified in 10 outbreaks and was the only serotype in 6. This serotype was identified in 1 of 21 confirmed ETEC outbreaks before 1996. PMID- 15109428 TI - Adherence barriers to antimicrobial treatment guidelines in teaching hospital, the Netherlands. AB - To optimize appropriate antimicrobial use in a university hospital and identify barriers hampering implementation strategies, physicians were interviewed regarding their opinions on antimicrobial policies. Results indicated that effective strategies should include regular updates of guidelines that incorporate the views of relevant departments and focus on addressing senior staff and residents because residents do not make independent decisions in a teaching-hospital setting. PMID- 15109429 TI - Neisseria meningitidis C:2b:P1.2,5 with intermediate resistance to penicillin, Portugal. AB - For 1 year, serogroup, serotype, serosubtype, and penicillin susceptibility of meningococci circulating in various regions in Portugal were evaluated. Most frequent phenotypes were B:4:P1.15 (13.4%) and C:2b:P1.2,5 (75.9%), which are also common in Spain. Overall, 27.5% of C:2b:P1.2,5 strains showed intermediate resistance to penicillin. Laboratory-based surveillance of meningococcal infection in Portugal provides important information to assess the adequacy of public health measures. PMID- 15109430 TI - Immunofluorescence assay for serologic diagnosis of SARS. AB - We evaluated an indirect immunofluorescence assay based on virus-infected cells for detecting anti-severe acute respiratory syndrome-associated coronavirus (SARS CoV) immunoglobulin (Ig) G antibody. All confirmed SARS cases demonstrated seroconversion or fourfold rise in IgG antibody titer; no control was positive. Sensitivity and specificity of this assay were both 100%. Immunofluorescence assay can ascertain the status of SARS-CoV infection. PMID- 15109431 TI - Tick-borne encephalitis in southern Norway. AB - The first five cases of human tick-borne encephalitis in Norway were reported from Tromoya, in Aust-Agder County. Serum specimens from 317 dogs in the same geographic area were collected. An enzyme immunoassay demonstrated antibody to human tick-borne encephalitis virus in 52 (16.4%) of the dogs, which supports the notion of an emerging disease. PMID- 15109432 TI - Endemic carbapenem-resistant Pseudomonas aeruginosa with acquired metallo-beta lactamase determinants in European hospital. AB - Acquired metallo-beta-lactamases (MBLs) can confer broad-spectrum beta-lactam resistance (including carbapenems) not reversible by conventional beta-lactamase inhibitors and are emerging resistance determinants of remarkable clinical importance. In 2001, multidrug-resistant Pseudomonas aeruginosa carrying bla(VIM) MBL genes were found to be widespread (approximately 20% of all P. aeruginosa isolates and 70% of the carbapenem-resistant isolates) at Trieste University Hospital. Clonal diversity and heterogeneity of resistance determinants (either bla(VIM-1)-like or bla(VIM-2)-like) were detected among MBL producers. This evidence is the first that acquired MBLs can rapidly emerge and establish a condition of endemicity in certain epidemiologic settings. PMID- 15109433 TI - Mycobacterium bovis infection, United Kingdom. AB - We describe the first documented spillover of bovine tuberculosis from animals into the human population of the United Kingdom since the resurgence of the disease in cattle in the country. This finding suggests that there may be a small risk for transmission to humans, making continued vigilance particularly necessary. PMID- 15109434 TI - Birth of healthy children after preimplantation diagnosis of beta-thalassemia. AB - BACKGROUND: Clinical programs for preventing beta-thalassemia are presently based on prospective carrier screening and prenatal diagnosis. This paper report an achievement of a pregnancy with unaffected embryos using in vitro fertilization and embryo transfer (IVF-ET), in combination with preimplantation genetic diagnosis (PGD), for a couple at risk of having children with beta-thalassemia. METHODS: A couple carrying different thalassemia mutations, both a codon 41 - 42 mutation and the IVS II 654 mutation, received standard IVF treatment, with intracytoplasmic sperm injection, embryo biopsiy, single cell polymerase chain reaction (PCR) and DNA analysis. Only unaffected or carrier embryos were transferred to the uterine cavity. After confirmation of pregnancy, a prenatal diagnosis was performed. RESULTS: Of a total of 13 embryos analyzed for beta globin mutations, PGD indicated that 2 were normal, 3 were affected, and 6 were carriers. Diagnosis could not be made in the other 2 embryos. Three embryos were transferred to the uterus on the third day after oocyte retrieval. Ultrasonography revealed a twin pregnancy with one blighted ovum. The prenatal genetic diagnosis revealed that both fetuses were unaffected, and two healthy boys were born, confirming the results of PGD. CONCLUSIONS: We developed a single cell based primer extension preamplification (PEP)-PCR assay for the detection of beta-thalassemia mutations. The assays were efficient and accurate at all stages of the procedure, and resulted in the birth of PGD-confirmed beta-thalassemia free children in China. PEP was used here in PGD for beta-thalassemia. PMID- 15109435 TI - Insulin-like growth factor binding protein 1 and human embryonic development during 6 - 10 gestational weeks. AB - BACKGROUND: Insulin-like growth factor binding protein-1 (IGFBP-1), which is a carrier of Insulin-like growth factors (IGFs) regulates the fetal development by working as an active factor controlling the combination of IGFs with their receptors. This study was designed to investigate the relationship between IGFBP 1 and human embryonic development during weeks 6 - 10 of gestation. METHODS: A total of 44 pregnant women with singleton pregnancy were divided into two groups: one with abnormal embryo development (n = 32) and the other with normal embryo development (n = 12). Enzyme-linked immunosorbent assay (ELISA) was employed to detect IGFBP-1 levels in maternal serum and decidual tissue. The expression of IGFBP-1 mRNA in deciduas was examined by reverse transcription polymerase chain reaction (RT-PCR) technique. RESULTS: The level of IGFBP-1 protein in maternal serum was significantly higher in the abnormal group [(125.36 +/- 47.93) micro g/ml] than in the normal group [(70.72 +/- 21.21) micro g/ml]. Both of IGFBP-1 and IGFBP-1 mRNA in deciduas were higher in abnormal group [(1.60 +/- 1.39) micro g/ml and 1.66 +/- 1.64, respectively] than in the normal group [(0.35 +/- 0.23) micro g/ml and 0.40 +/- 0.20, respectively]. The level of IGFBP-1 in maternal serum was positively correlated with IGFBP-1 mRNA (r = 0.90, P < 0.05) and IGFBP 1 protein (r = 0.92, P < 0.05) in decidual tissue. CONCLUSIONS: During weeks 6 - 10 of gestation, abnormal embryonic development is correlated with elevated IGFBP 1. The level of IGFBP-1 in maternal serum is related to the concentrations of IGFBP-1 mRNA and IGFBP-1 in decidual tissue. The IGFBP-1 level in maternal serum may be used as a predictive marker to evaluate embryonic development. PMID- 15109436 TI - Association of HLA-DQB1 coding region with unexplained recurrent spontaneous abortion. AB - BACKGROUND: DNA analysis has shown a lack of significant compatibility between couples affected by unexplained recurrent spontaneous abortion (URSA) compared with normal fertile couples, [8] although one study that made use of a PCR sequence-specific oligonucleotide (SSO) method did observe evidence of significant compatibility in the HLA-DQA1 and DQB1 alleles between patients and aborted fetuses. [9] This study was designed to investigate whether URSA were associated with particular DQ alleles or promoter alleles. METHODS: Thirty-two patients with URSA and 54 women who had had at least one successful pregnancy were included in this study. HLA-DQ genotyping was performed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. The HLA-DQB1 promoter was detected by the SSO and sequence-specific primer (SSP) methods. The DQA1, DQB1, and DQB1 promoter (QBP) gene frequencies in the patients were compared with the gene frequencies in normal controls. The data were analyzed statistically with the chi(2) and Fisher's exact tests. RESULTS: The results showed that the frequency of DQB1 * 0604/0605 was significantly higher and the frequency of DQB1 * 0501/0502 was significantly lower in the patient group as compared with the normal controls. In addition, the frequencies of the DQA1 * 01-DQB1 * 0604/0605 and QBP6.2-DQB1 * 0604/0605 haplotypes were overrepresented in the patients relative to the controls. Our results did not show any differences between URSA patients and the controls with regard to DQA1 and QBP allele frequencies. CONCLUSIONS: Our data suggest that URSA is associated with the HLA-DQB1 coding region, and is not associated with its upstream regulatory region. The DQB1 * 0604/0605, DQA1 * 01-DQB1 * 0604/0605, and QBP6.2 DQB1 * 0604/0605 haplotypes may confer susceptibility to URSA, while the DQB1 * 0501/0502 allele may protect women from URSA. PMID- 15109437 TI - Effect of estrogen deprivation on follicle/oocyte maturation and embryo development in mice. AB - BACKGROUND: It is believed that estrogen plays pivotal roles in the regulation of follicle/oocyte maturation and oocyte fertilizability. It is also involved in the functional preparation of the fallopian tubes for subsequent gamete interaction, in early embryonic development occurring in the tubal microenvironment, and in the preparation of the uterus for implantation. This study was designed to determine whether estrogen is required for follicular and embryonic development. METHODS: The biosynthesis of estrogen was blocked by a daily injection of the aromatase inhibitor, Arimidex, at a dose of 100 micro g/d, using 3 - 4 week old C57B6 F1 female mice. Injections were continued for 3 days in experiment 1 (n = 10) and for 5 days in experiment 2 (n = 23). Mice in the control group (n = 27) were given the same amount of saline. Exogenous gonadotrophin [7.5 IU pregnant mare serum gonadotrophin (PMSG)] was administered to induce follicular growth and development on the second day. In experiment 1, we tested estrogen and progesterone levels and examined ovary morphology two days later. In experiment 2, 47 hours after PMSG injection, 5 IU human chorionic gonadotropin (hCG) was given and two female mice were then caged with a male mouse overnight. Two days later, we measured estrogen and progesterone levels. We then removed the embryos, cultured them, and examined embryonic development every 24 hours for 3 days. RESULTS: Before hCG injection, estrogen levels in mice from the Arimidex group were suppressed by 94%, and progesterone levels were suppressed by 75%. There was no difference between the two groups in mean number of total follicles found per animal (30.4 follicles/animal in the control group and 27 follicles/animal in the Arimidex group). Two days after hCG injection, estrogen levels in the Arimidex group were significantly lower than that in the control group (P < 0.01), while progesterone levels were not significantly lower (P > 0.05). The rate of development of embryos, morulae, blastocysts, and hatching blastocysts was not significantly different between the two groups (P = 0.20, 0.10, 0.44, and 0.38, respectively). CONCLUSIONS: In the present study, by depriving mice of normal estrogen support, we have been able to rule out the absolute need for rising levels of estrogen for the completion of the follicular maturation process and the development of embryos in vitro. PMID- 15109438 TI - Investigation of the frequency of chromosomal aneuploidy using triple fluorescence in situ hybridization in 12 Chinese infertile men. AB - BACKGROUND: Chromosomal aberrations are the major cause of pre- and post implantation embryo wastage and some studies suggest that half of all human conceptions have a chromosomal abnormality. A chromosomal aberration in human sperms is also one of the causes of failure of in vitro fertilization. This study was designed to ascertain whether chromosomal aneuploidy in spermatozoa is a risk factor for male infertility. METHODS: Twelve infertile men were divided into two groups: 10 with oligoasthenoteratozoospermia (OAT, Group A) and two with a normal semen analysis (Group B). Two normal healthy sperm donors acted as controls (Group C). We used fluorescence in situ hybridization (FISH) and probes for chromosomes X, Y and 18 to determine the frequency of aneuploidy. RESULTS: The frequencies of spermatozoa disomy for chromosomes X, Y and 18 were 0.30% and 0.30%, respectively, in Group B. The percentages were not significantly different from those of Group C (0.15% and 0.16%). The frequencies of nullisomy for chromosomes X, Y and 18 were 0.15% and 0 for Group B, and 0 and 0.15% for Group C (P > 0.05). In Group A, the incidences of disomy were 1.13% and 0.96% and the frequencies of nullisomy were 1.13% and 1.60%. In these three groups, the incidences of diploidy were 0.60%, 1.00%, and 0.30%, respectively. Both the frequencies of disomic and nullisomic spermatozoa for chromosomes X, Y, and 18 and of diploid spermatozoa were significantly higher in Group A than in Groups B and C. The estimated total aneuploidy rates in the sperm from the three groups were 42.44%, 6.05%, and 2.59%, respectively. CONCLUSION: These results indicate that chromosomal aneuploidy in spermatozoa may be a risk factor for infertility. PMID- 15109439 TI - Comparison of three methods for the gene analysis of fetal cells from maternal peripheral blood. AB - BACKGROUND: Although great advances in techniques for noninvasive prenatal diagnosis using fetal cells from maternal peripheral blood have achieved, current technology does not meet the demands required for clinical use. In this study, we aimed to establish reliable methods for the gene analysis of fetal cells from maternal peripheral blood. METHODS: Primed extension preamplification (PEP) polymerase chain reaction (PCR), multiple primed in situ labeling (PRINS), and nested PCR were individually applied to detect the sex determining region Y (SRY) gene in single fetal cells collected from maternal peripheral blood. RESULTS: The sensitivity and specificity of the detection of the SRY gene by PEP-PCR were 97.39% (149/153) and 99.17% (119/120), respectively. The sensitivity and specificity of PRINS were 97.56% (40/41) and 100% (35/35), respectively. The sensitivity and specificity of nested-PCR were 80.00% (24/30) and 87.50% (14/16), respectively. CONCLUSIONS: PEP-PCR and PRINS are reliable techniques for the gene analysis of single fetal cells from maternal peripheral blood because of their high sensitivity and specificity. PEP-PCR and PRINS can be used as standard methods of noninvasive prenatal diagnosis using single fetal cells from maternal peripheral blood. PMID- 15109440 TI - Apparent protective effect of high density lipoprotein against coronary heart disease in the elderly. AB - BACKGROUND: This study was designed to evaluate the relationship between high density lipoprotein cholesterol (HDL-C) level and acute myocardial infarction (AMI) and coronary heart disease (CHD) death and to explore the protective effect of HDL against CHD in the elderly Chinese. METHODS: Started from 1986, 1211 retirees (92% males) were enrolled consecutively and studied prospectively. The average starting age was 70 +/- 9 years, and that at the end of the study was 80 +/- 9 years. During the follow-up study, all the participants received yearly physical examination and blood chemistry survey from 1986 - 2000. The average duration of the follow up study was 11.2 years. The end point of this study was either attacks of AMI or death due to CHD and other causes. CHD risk factors were screened by logistic regression analysis. According to their HDL-C levels, cases were divided into low (< 1.03 mmol/L), medium (or normal, 1.03 - 1.56 mmol/L) and high (> 1.56 mmol/L) level groups, the differences in incidence of AMI and CHD death in each group were analyzed. RESULTS: The cumulative attacks of acute coronary syndrome (mostly AMI) were 214 cases, including 89 cases of coronary death and 308 death caused by other diseases during the follow up study. AMI occurrence and CHD death in normal HDL-C group were lower than those in the low HDL-C group by 40% and 53%; and those in the high HDL-C group were lower than in the normal group by 56% and 50%, respectively. Statistical analysis on normal lipid cases (411 cases, total cholesterol < 5.17 mmol/L, triglyceride < 1.69 mmol/L) revealed that the cases at low HDL-C level had similar rates of AMI events and CHD mortality as those of the entire group (including hyperlipidemia); however, AMI attacks and CHD deaths decreased significantly at the normal and high HDL-C levels. The results demonstrated that the protective effect of HDL against coronary artery disease is more prominent in people with low lipid level. CONCLUSION: Low HDL is an important independent risk factor for AMI attacks and CHD death in the elderly; high HDL has significant protective effect against coronary artery disease. PMID- 15109441 TI - Effects of right atrial and ventricular DDD pacing on cardiac function and ventricular contraction synchrony. AB - BACKGROUND: Right ventricular apical pacing has been reported to reduce cardiac performance. But there are few reports on the effects of dual chamber (DDD) pacing on cardiac function compared to sinus rhythm. In this study, we evaluated the effects of right atrial and ventricular DDD pacing on cardiac function and ventricular contraction synchrony using equilibrium radionuclide angiography. METHODS: Ten patients implanted with a right atrial and ventricular DDD pacemaker underwent equilibrium radionuclide angiography. The scintigraphic data were obtained during sinus rhythm and pacing rhythm. Cardiac function parameters were obtained semimanually. Phase analysis was used to study the ventricular activation sequence and ventricular synchrony. RESULTS: The left ventricular 1/3 ejection fraction decreased significantly during pacing compared with that during sinus rhythm [(23.4 +/- 6.1)% vs (27.7 +/- 4.5)%, P = 0.01]. Regional ejection fraction also decreased during pacing, although the difference was not statistically significant. Phase analysis showed that the right ventricle was activated earlier than the left ventricle during pacing, and that the phase shift was significantly greater during pacing than that during sinus rhythm [64.13 degrees +/- 16.80 degrees vs 52.88 degrees +/- 9.26 degrees, P = 0.007]. The activation of both ventricles occurred simultaneously during sinus rhythm, with the activation sequence from proximal septum or base of left ventricle to apex. The earliest activation during pacing occurred at the right ventricular apex, and subsequently spread to the base and left ventricle. CONCLUSION: Right atrial and ventricular DDD pacing impairs left ventricular systolic function and ventricular synchrony. PMID- 15109442 TI - Identification of syndrome X using intravascular ultrasound imaging and Doppler flow mapping. AB - BACKGROUND: The purpose of this study was to assess the morphological changes and physiological function of coronary arteries in patients presenting with chest pain but having normal coronary angiograms, using intravascular ultrasound imaging (IVUS) and intracoronary Doppler (ICD) flow measurements, in order to elucidate the mechanism of syndrome X. METHODS: A total of 126 patients [67 males, 59 females, mean age (53.1 +/- 13.0) years] who experienced chest pain but had normal coronary angiograms were included in this study. ICD flow measurements of the left anterior descending coronary artery (LAD) were performed using a Cardiometrics FloMap II system. Coronary flow velocity reserve (CFVR) was defined as the ratio of the average peak velocity during hyperemia to that at baseline, induced by an intracoronary bolus injection of 18 microg adenosine. A 3.2F or 2.9 F 30 MHz mechanical rotating ultrasound catheter (CVIS, Boston Scientific) or a 3.0 F 20 MHz electronic ultrasound catheter (Endosonics) was used for IVUS. RESULTS: The mean CFVR value of the LAD was 2.71 +/- 0.74. Reduction of CFVR (< 3.0) was found in 82 of 126 (65.1%) patients. IVUS images of the LAD were available for 109 patients. Plaque formation was detected in 76/109 (69.7%) patients. Based on the presence or absence of plaque formation as well as the reduction or non-reduction of CFVR, patients were divided into four groups: Group I (n = 10), normal IVUS findings and normal CFVR; Group II (n = 23), normal IVUS findings with reduction in CFVR; Group III (n = 29), IVUS evidence of plaque formation but normal CFVR; and Group IV (n = 47), IVUS evidence of plaque formation with reduction in CFVR. CONCLUSION: This study shows the important clinical value of a combination of IVUS and ICD in diagnosing patients with angiographically normal coronary arteries. Only 10% of patients studied (Group I) were found to be truly free of coronary disease, while 20% of patients (Group II) would be diagnosed as suffering from syndrome X. PMID- 15109443 TI - Electrical heterogeneity of canine right ventricular transient outward potassium currents. AB - BACKGROUND: Some studies have confirmed that the right ventricular walls of most rodents, such as canines and humans, have evident transient outward potassium current (Ito1) heterogeneity, and this heterogeneity is closely related to J point elevation, J wave formation, and some ventricular tachycardias such as ventricular fibrillations caused by Brugada syndrome. This study is designed to investigate transmural electrical heterogeneity of the canine right ventricle during repolarization (phase 1) from the viewpoint of 4-aminopyridine sensitive and calcium-independent Ito1. METHODS: Adult canine single right ventricular epicardial (Epi) cells, mid-myocardial (M) cells, and endocardial (Endo) cells were enzymatically dissociated. Whole cell voltage-clamp recordings were made to compare the Ito1 values of the three cell types. RESULTS: At 37 degrees C and using 0.2 Hz and +70 mV depolarizing test potentials, the average peak Ito1 values of Epi cells and M cells averaged (4070 +/- 1720) pA and (3540 +/- 1840) pA, respectively. The activated and inactivated Epi and M cells kinetic processes were in accordance with the Boltzmann distribution. Compared with Ito1 in Epi cells and M cells, the average peak Ito1 in Endo cells was very low, averaged (470 +/- 130) pA. CONCLUSIONS: These results suggest that there are evident differences and potent gradients in Ito1 between the three cardiac cell types, especially between Epi and Endo cells. These differences are among the prominent manifestations of right ventricular electrical heterogeneity, and may form an important ionic basis and prerequisite for some malignant arrhythmias in the right ventricle, including those arising from Brugada syndrome and other diseases. PMID- 15109444 TI - Morphological and electrophysiological study on the inferior nodal extension and transitional cellular band in the rabbit atrioventricular junctional area. AB - BACKGROUND: Advances in catheter ablation procedures for the treatment of supraventricular arrhythmias have created the need to understand better the morphological and electrophysiological characteristics of the inferior nodal extension (INE) and transitional cellular band (TCB) in the atrioventricular (AV) junctional area. METHODS: Firstly, we observed the histological features of 10 rabbit AV junctional areas by serial sections under light microscopy. Then we recorded the action potentials (APs) of transitional cells (TCs) in the INE, TCBs, AV node, and ordinary right atrial myocytes from the AV junctional area of 30 rabbits using standard intracellular microeletrode techniques. RESULTS: Under light microscopy, the INE appeared to be mostly composed of transitional cells linking upward to the AV node. Four smaller TCBs originated in the orifice of the coronary sinus, the region between the septal leaflet of the tricuspid valve and the coronary sinus, the inferior wall of the left atrium, and the superior interatrial septum, respectively, all linking to the INE or the AV node. Compared with ordinary atrial myocytes, the AP of the TCs in both the INE and the TCBs had a spontaneous phase 4 depolarization (not present in ordinary atrial myocytes), with a less negative maximum diastolic potential, a smaller amplitude, a slower maximum velocity of AP upstroke, and a longer action potential duration at 50% repolarization (APD50) and at 30% repolarization (APD30). The AP characteristics of these TCs were similar to those of the AV node, except that the velocities of the phase 4 spontaneous depolarization were slower and their action potential durations at 90% repolarization (APD90) were shorter. Moreover, APD50 and APD30 of the TCs of the TCBs were shorter than in the case of TCs of the AV node. CONCLUSIONS: The TCs of the INE and TCBs are similar to slow response automatic cells. They provide a substrate for slow pathway conduction. In addition, repolarization heterogeneity exists in the AV junctional area. PMID- 15109445 TI - Experimental study on the mechanism of sex difference in the risk of torsade de pointes. AB - BACKGROUND: Torsade de pointes (TdP) is a form of polymorphic ventricular tachycardia featuring prolonged QT intervals. Female gender is associated with an increased risk of TdP. However, the causes of the sex difference in risk are poorly understood. Recently, transmural dispersion of repolarization (TDR) has been implicated in the genesis of TdP. Consequently, we compared TdP incidence and TDR between male and female rabbit hearts in order to investigate the mechanism of sex difference in TdP risk in rabbits in vitro. METHODS: By means of monophasic action potential recording techniques, the monophasic action potential of the epicardium, midmyocardium, and endocardium were simultaneously recorded using specially designed plunge-needle electrodes placed across the left ventricular free wall of both female (n = 8) and male (n = 8) rabbit hearts purfused by the Langendorff method. TdP was induced by bradycardia, d-sotalol, and low-K+, Mg2+ Tyrode solution. RESULTS: TDR measurements in all three myocardial layers of male and female rabbit hearts were (18 +/- 2) ms and (21 +/- 2) ms, respectively (n = 8, P > 0.05). After perfusion with d-sotalol, the 90% monophasic action potential duration was prolonged in both male and female rabbits. TDR in male and female rabbit hearts increased to (29 +/- 2) ms and (61 +/- 2) ms, respectively, a difference that is significant. Eight female rabbit hearts had early afterdepolarization and 7 of them developed TdP. Seven male rabbit hearts had early after depolarization, but only one of these hearts developed TdP. CONCLUSION: Greater TDR may play an important role in the higher incidence of TdP in female rabbit hearts. PMID- 15109446 TI - Dietary hypercholesterolemia aggravates contrast media-induced nephropathy. AB - BACKGROUND: Contrast media administration can result in severe nephrotoxicity under pathological conditions such as diabetic nephropathy, congestive heart failure, dehydration, et al. The purpose of this study was to evaluate the effects of dietary hypercholesterolemia on contrast media-induced changes in renal function, blood flow, and histopathology. METHODS: Rats were fed either on a normal rodent diet (group N) or a high-cholesterol supplemented diet (group H; 4% cholesterol and 1% cholic acid) for 8 weeks. Half of the animals (n = 6) from each diet group were then given a tail vein injection of 60% diatrizoate (6 ml/kg; group NC and group HC) and the other half were administered saline. Total serum cholesterol, triglyceride, serum creatinine, creatinine clearance rate, fractional excretion of sodium and potassium, and cortical nitric oxide production were determined one day following contrast media administration. Renal blood flow was determined by color Doppler flow imaging and pulsed-mode Doppler. Renal histopathology was observed by light microscopy. RESULTS: Total serum cholesterol and resistance indices of renal blood vessels increased significantly, while creatinine clearance rate and production of nitric oxide in the renal cortex decreased markedly in group HC and group H when compared to group N and group NC. The creatinine clearance rate decreased significantly in group HC compared to group H. Serum creatinine levels and fractional excretion of sodium and potassium in group HC were significantly higher than those in the other three groups. Severe tubular degeneration and necrosis, protein cast accumulation, and medullary congestion were found in group HC. CONCLUSION: Hypercholesterolemia is a risk factor for contrast media-induced nephropathy. Hypercholesterolemia aggravates contrast media-induced nephrotoxicity through the reduced production of nitric oxide. PMID- 15109447 TI - Effect of irbesartan on angiotensin II-induced hypertrophy of human proximal tubular cells. AB - BACKGROUND: Intrarenal activation of the renin angiotensin system (RAS) plays an important role in mediating renal fibrosis. Both angiotensin converting enzyme inhibitors (ACEIs) and angiotensin II (AngII) receptor antagonists have been shown to exert a protective role against diabetic and non-diabetic nephropathy. However, the exact mechanism of how blocking local RAS prevents renal fibrosis is unclear. The present study was to investigate the influence of a new AngII receptor antagonist, irbesartan (Irb), on AngII-induced hypertrophy in human proximal tubular cell line (HK-2). METHODS: The cell line, HK-2, was grown in Dulbeccos's Modified Eagle's Medium containing 10% heat-inactivated fetal calf serum. After rested in serum-free medium for 24 hours, the effects of Irb on AngII (10(-7) mol/L)-induced [(3)H]-leucine incorporation, total protein content (measured by the Coomassie brilliant blue G250 method), and change in cell size (determined by scanning electron microscopy) were observed. The influence of Irb on the cell cycle was analyzed by fluorescence activated cell sorter (FACS) flow cytometry. RESULTS: AngII induced cell hypertrophy in a time and dose dependent manner. Stimulation of cells with AngII for 48 hours resulted in a increase in [(3)H]-leucine incorporation [0 hour: (5584 +/- 1016) cpm/10(5) cells vs 48 hours: (10741 +/- 802) cpm/10(5) cells, P < 0.05], which was significantly attenuated by treatment with Irb. AngII significantly increased the total protein content in HK-2 cells [control: (0.169 +/- 0.011) mg/10(5) cells vs AngII group: (0.202 +/- 0.010) mg/10(5) cells, P < 0.05], which was also markedly inhibited by cotreatment with Irb (P < 0.01). Scanning electron microscopy showed that AngII induced an increase in average physical cell size, which was significantly inhibited by Irb [control: (11.92 +/- 1.62) microm; AngII group: (20.63 +/- 3.83) micro m; AngII + Irb group: (13.59 +/- 3.15) micro m; P < 0.01 vs control, respectively]. Furthermore, flow cytometry revealed that AngII arrested cells in the G(0)-G(1) phase, which was significantly reversed by treatment with Irb [G(0) G(1) cells in AngII group: (76.09 +/- 1.82)%, in AngII + Irb group: (67.00 +/- 2.52)%, P < 0.05]. CONCLUSION: Irb can inhibit AngII-induced hypertrophy in HK-2 cells. PMID- 15109448 TI - Genetic study of a large Chinese kindred with von Hippel-Lindau disease. AB - BACKGROUND: Von Hippel-Lindau (VHL) disease is a heraditary cancer syndrome caused by germline mutations of the VHL tumor on the suppressor gene. This study was to show the clinical characteristics of a large Chinese kindred with von Hippel-Lindau disease and to evaluate the role of the genetic test of VHL disease in the diagnosis of VHL disease and clinical screening of members of the VHL disease family. METHODS: DNA extracted from peripheral blood was amplified by PCR to three exons of the VHL gene in 27 members of a large kindred with VHL disease. PCR products were directly sequenced. The involvements of multi-organs in the kindred with VHL disease were confirmed by history taking and radiography. RESULTS: Of 47 members in the four generations of the kindred, 18 members were diagnosed as having VHL disease. Clinical manifestations of 18 patients included: central nervous system (CNS) hemangioblastoma (5), renal cell carcinoma and CNS hemangioblastoma (3), renal cell carcinoma and retinal angioma (3), renal cell carcinoma and multiple pancreatic cysts (1), renal cell carcinoma and retinal angioma and multiple pancreatic cysts (2), renal cell carcinoma and CNS hemangioblastomas and multiple pancreatic cysts (1), and multiple pancreatic cysts and multiple renal cysts (1), multiple pancreatic cysts (2). The common lesions of the 18 patients were renal cell carcinoma (55.6%), CNS hemangioblastoma (50.0%), retinal angioma (27.8%), and multiple pancreatic cysts (38.9%). Among the 27 members who volunteered for genetic analysis, 15 members including 9 affected family patients and 2 asymptomatic patients and 4 carriers, who are still alive, presented a codon 78 from Asn to Ser change at nucleotide 446 (A-->G) in exon 1. Four members were carriers with the same VHL gene mutation. Two asymptomatic patients were initially diagnosed by genetic testing and subsequently confirmed radiologically and surgically. Members without gene mutation had no clinical evidence of VHL disease. CONCLUSIONS: The large Chinese kindred with VHL disease was classified as type I. The main characteristics in the kindred were higher incidence of renal cell carcinoma and lower incidence of retinal angioma. Genetic test plays an important role in early detecting asymptomatic patients and the carriers in clinical screening of members of the families with VHL disease. It is also important to prevent the transmission of VHL disease to their offsprings in the kindred. PMID- 15109449 TI - Polymorphism of the leptin gene promoter in pedigrees of type 2 diabetes mellitus in Chongqing, China. AB - BACKGROUND: It has been shown that the presence of leptin is associated with deabefes, glucose wefabolism and insulin metablism. In this research, we evaluated the presence of the leptin C-2549-A polymorphism in the Chinese population in Chongqing and verified its association with plasma leptin levels and anthropometric, metabolic, and clinical parameters. METHODS: Two hundred and sixty-nine patients with diabetes, 135 non-diabetic first-degree relatives of the patients, and 85 healthy controls were screened for the presence of C-2549-A polymorphism using a PCR-RFLP assay. Body mass index, fasting leptin, fasting insulin, fasting glucose and homeostatic model assessment for insulin resistance (HOMA)-IR were also determined. RESULTS: In the type 2 diabetes group, AA genotype frequency (6.32%) and A allele frequency (34.94%) was higher than in normal controls (1.18% and 25.29%, respectively). Diabetic patients with the AA genotype had lower fasting leptin and insulin levels than those with other genotypes. Carriers with the AC genotype had decreased fasting leptin and insulin levels and longer duration of disease as compared with those with CC genotype. The HOMA-IR of patients with AA or AC genotypes was lower than those with the CC genotype. In non-diabetic relatives group, individuals with the AA genotype had a lower fasting leptin level than those with the AC genotype. The fasting insulin and HOMA-IR level of carriers of the AA or AC genotype were lower than those of the CC genotype. CONCLUSION: The C-2549-A polymorphism in the leptin gene is associated with fasting leptin in patients with type 2 diabetes. The distribution of the genotypes in diabetic subjects from diabetic pedigrees differs from those in normal controls. The A allele frequency in diabetic patients is higher than that in normal controls. The haplotypes defined by genotypes are different in the familial subjects. PMID- 15109450 TI - Construction of adeno-associated virus coexpression system for human angiopoietin 1 and VEGF gene. AB - BACKGROUND: Ischemic disease is one of the leading causes of death in the world. In order to further study gene therapy for ischemic disease, we constructed a recombinant plasmid for co-expression of human angiopoietin-1 and vascular endothelial growth factor 165(VEGF165) gene in adeno-associated virus (AAV) gene delivery system. METHODS: Human angiopoietin 1 and VEGF165 gene were obtained using PCR. The upstream of angiopoietin 1 contained restriction enzyme site HindIII, and the downstream of angiopoietin 1 contained restriction enzyme site BamHI. The upstream of VEGF165 contained restriction enzyme site BglII, and the downstream of VEGF165 contained restriction enzyme site BamHI. Using the multiple cloning sites (MCS) in plasmid pZero++ such as BamHI, BglII, HindIII, NotI, XhoI, XbaI, SalI, BspHI, KspI and the corresponding MCS in plasmid pAAV-MCS, angiopoietin 1 and VEGF165 gene were subcloned into pAAV-MCS. RESULTS: DNA sequencing revealed that the PCR- amplified angiopoietin 1 and VEGF165 were consistent with NCBI Gene Bank. The recombinant plasmid was identified using PCR and digestion, which proved to be consistent with our hypothesis. In recombinant plasmid, angiopoietin1 and VEGF possessed a CMV promoter and polyA terminator system respectively, thus assuring co-expression of the two genes. CONCLUSION: Successful construction of AAV co-expression system for human angiopoietin 1 and VEGF165 gene will provide the foundation for gene therapy to cure severe ischemic disease. PMID- 15109451 TI - Peptide nucleic acids arrest the growth of gastric cancer cells SGC7901. AB - BACKGROUND: Peptide nucleic acid (PNA) has many characteristics useful in molecular biology. This paper described an effective way to raise the cell ingestion rate of PNA so as to kill gastric cancer cells. METHODS: Heteroduplexes of PNAs and oligonucleotides, wrapped by Lipofectamine 2000, were used to infect SGC7901 cells. The inhibitive effect of heteroduplexes was evaluated by analyzing cell clone forming and cell growth rate. Telomerase activity of SGC7901 cells was detected by polymerase chain reaction enzyme-linked immunosorbent assay (PCR ELISA) and silver staining assay. RESULTS: PNAs showed a dose-dependent inhibition of cell proliferation. The percentage of proliferation inhibition was 99.4% after 7 days; the rate of cloning inhibition was 98.2% after 8 days; whereas for oligonucleotide groups, at the same concentration, the percentages were 50.1% and 67.5% respectively. Antisense PNA-DNA-Lipofectamine 2000 group (AP D-L group) exhibited significantly different percentages from the control groups (P < 0.05). The test result indicated that telomerase activity of the AP-D-L group was inhibited (P < 0.05). At the same time, the impact on cell morphology was observed. CONCLUSIONS: The results showed that PNAs are potent antisense reagents. The telomerase-associated therapies are very promising for the treatment of malignant tumours. PMID- 15109452 TI - Antioxidation of melatonin against spinal cord injury in rats. AB - BACKGROUND: The iron catalyzed lipid peroxidation plays an important role in the autodestruction of the injured spinal cord. This study was to detect the antioxidation of melatonin against spinal cord injury (SCI) in rats. METHODS: Sity Sprague-Dawley rats were randomly divided into four groups: group A (n = 15) for laminectomyanly, group B (n = 15) for laminectomy with SCI, group C (n = 15) for SCI and intraperitoneal injection of a bolus of 100 mg/kg melatonin, and group D (n = 15) for SCI and intraperitoneal injection of saline containing 5% ethanol. The SCI of animal model was made using modified Allen's method on T12. Six rats of each group were sacrificed 4 hours after injury, and the levels of free iron and malondialdehyde (MDA) of the involved spinal cord segments were measured by the bleomycin assay and thiobarbituric acid (TBA) separately. Functional recovery of the spinal cord was assessed by Modified Tarlov's scale and the inclined plane method at 1, 3, 7, 14, 21 days after SCI. The histologic changes of the damaged spinal cord were also examined at 7 days after SCI. RESULTS: After SCI, the levels of free iron and MDA were increased significantly and the modified Tarlov's score and inclined plane angle decreased significantly in groups B and D. In group C, the Tarlov's score and inclined plane angle were increased significantly at 7, 14 and 21 days, with histological improvement. CONCLUSION: Melatonin can reduce the level of lipid peroxidation and prevent damage to the spinal cord of rat. PMID- 15109453 TI - Comparative study on seeding methods of human bone marrow stromal cells in bone tissue engineering. AB - BACKGROUND: In general the traditional static seeding method has its limitation while the dynamic seeding method reveals its advantages over traditional static method. We compared static and dynamic seeding method for human bone marrow stromal cells (hBMSCs) in bone tissue engineering. METHODS: DNA assay was used for detecting the maximal initial seeding concentration for static seeding. Dynamic and static seeding methods were compared, when scaffolds were loaded with hBMSCs at this maximal initial cell seeding concentration. Histology and scanning electron microscope (SEM) were examined to evaluate the distribution of cells inside the constructs. Markers encoding osteogenic genes were measured by fluorescent RT-PCR. The protocol for dynamic seeding of hBMSCs was also investigated. RESULTS: DNA assay showed that the static maximal initial seeding concentration was lower than that in dynamic seeding. Histology and SEM showed even distribution and spread of cells in the dynamically seeded constructs, while their statically seeded counterparts showed cell aggregation. Fluorescent RT-PCR again showed stronger osteogenic potential of dynamically seeded constructs. CONCLUSION: dynamic seeding of hBMSCs is a promising technique in bone tissue engineering. PMID- 15109454 TI - Inflammatory reaction after focal cerebral ischemia in mouse. AB - BACKGROUND: In response to the inflammatory reaction, circulating leukocytes aggregate and adhere to the endothelial cells and eventually pervade into tissues, resulting in cell damage. This study was to detect the inflammatory reactions in mouse focal cerebral ischemia and their distinct characteristics in the ischemic basal ganglia and surrounding cortex. METHODS: Mice were subjected to permanent occlusion of the left middle cerebral artery (MCAO) by introducing a suture for 2 to 120 hours. The expression of intercellular adhesion molecule 1 (ICAM-1) and Mac-1 was determined immunohistochemically. The myeloperoxidase (MPO) activity of the ischemic regions was measured. RESULTS: Four hours after MCAO, the number of ICAM-1 positive vessels in the ischemic basal ganglia increased (9.2 +/- 2.8 per mm(2)), peaked at 48 hours (29.6 +/- 4.8 per mm(2)), and decreased after 72 hours. In the ischemic cortex, the number increased rapidly 4 hours after MCAO (19.4 +/- 6.1 per mm(2)), peaked at 48 hours (44.4 +/- 16.8 per mm(2)), and declined after 72 hours. Mac-1 positive cells were seen in the ischemic basal ganglia (3.4 +/- 1.2 per mm(2)) 12 hours after MCAO, peaked after 48 hours (20.2 +/- 6.3 per mm(2)), and decreased after 72 hours. In the ischemic cortex, however, the number increased 4 hours after MCAO (4.3 +/- 1.7 per mm(2)), peaked after 48 hours (20.9 +/- 8.4 per mm(2)), and remained high at 120 hours. The MPO activity increased in the ischemic basal ganglia 12 hours after MCAO (0.111 +/- 0.023 U/g), peaked after 24 hours (0.194 +/- 0.059 U/g), and decreased after 72 hours. In the ischemic cortex, the MPO activity increased 12 hours after MCAO (0.110 +/- 0.032 U/g), peaked after 24 hours (0.210 +/- 0.067 U/g), and remained elevated at 120 hours. CONCLUSIONS: The increased expression of ICAM-1 in the ischemic brain of mouse in the early phase of MCAO followed by the over-expression of Mac-1 and the increased MPO activity suggests that focal ischemia leads to early onset of inflammation. The inflammatory response is more persistent and intensive in the ischemic cortex than in the ischemic basal ganglia. PMID- 15109455 TI - Effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD(34+) hematopoietic cells in bone marrow of asthmatic mice. AB - BACKGROUND: Corticosteroids remain the most effective therapy available for asthma. They have widespread effects on asthmatic airway inflammation. However, little is known about the effects of corticosteroids on the production of bone marrow inflammatory cells in asthma. This study observed the effects of glucocorticoid and cysteinyl leukotriene 1 receptor antagonist on CD34+ hematopoietic cells, so as to explore the possible effectiveness of a bone marrow targeted anti-inflammatory strategy. METHODS: Balb/c mice were sensitized and challenged with ovalbumin (OVA) to establish an asthmatic model. For two consecutive weeks, asthmatic mice were challenged with OVA while being given either prednisone, montelukast, prednisone plus montelukast, or sterile saline solution. The mice were killed 24 hours after the last challenge with OVA, and bronchoalveolar lavage fluid (BALF), peripheral blood, and bone marrow were collected. Eosinophils in peripheral blood and BALF, and nucleated cells in BALF, peripheral blood, and bone marrow were counted. The percentages of CD34+ cells, CD4+ T lymphocytes and CD8+ T lymphocytes among nucleated cells in peripheral blood and bone marrow were counted by flow cytometry. Immunocytochemistry and in situ hybridization were employed to detect expression of CD34 and interleukin (IL)-5Ralpha mRNA (CD34+ IL-5Ralpha mRNA+ cells) among bone marrow hematopoietic cells. RESULTS: Compared with the sterile saline solution group, the number of eosinophils in BALF and peripheral blood, CD34+ cells in peripheral blood and bone marrow, and CD34+ IL-5Ralpha mRNA+ cells in bone marrow of mice from the prednisone and prednisone plus montelukast groups were significantly lower (P < 0.01). The number of eosinophils in BALF from the montelukast group was also significantly lower (P < 0.05). CONCLUSIONS: The results suggest that, in this asthmatic mouse model, prednisone probably inhibits proliferation, differentiation, and migration of CD34+ cells in bone marrow, blocks eosinophilopoiesis in bone marrow, and interferes with eosinophil migration into peripheral blood and subsequent recruitment in the airway. In addition, montelukast may suppress eosinophil infiltration into the lungs of asthmatic mice. However, a significant inhibitory effect of montelukast on the proliferation and migration of CD34+ cells and a cooperating effect with prednisone on bone marrow of asthmatic mice were not observed. PMID- 15109456 TI - Etiological analysis on ocular fungal infection in the period of 1989 - 2000. AB - BACKGROUND: This study was to review the distribution and shifting trend of fungal of culture specimens isolated from eyes of patients at the Beijing Institute of Ophthalmology, Tongren Hospital, Beijing, China. METHODS: The fungal culture-positive rate, the distribution and change of isolates of 2609 specimens collected in a 12-year period (1989 - 2000) were retrospectively analyzed. RESULTS: In 775 positive cultures, 707 specimens (91.2%) were from the cornea, 22 (2.8%) from the conjunctiva, 15 (1.9%) from the anterior chamber, 9 (1.2%) from the vitreous body, 3 (0.4%) from the lacrimal sac, and 19 (2.5%) from other parts of the eye. The average culture-positive rate was 29.7%. The ratio of the positive cultures in the first half year (from January to June) to those in the second half (from July to December) was 1:2.1. The main genus cultured was Fusarium sp (58.7%), followed by Aspergirum sp (16.8%). The percentage of Fusarium sp was increased from 53.6% (1989 - 1994) to 60.2% (1995 - 2000), whereas the percentage of Aspergirum sp was decreased from 22.3% (1989 - 1994) to 15.1% (1995 - 2000). CONCLUSIONS: Fusaruim sp is one of the most predominant pathogens of ocular fungal infection in northern China and its incidence tends to increase, but that of Aspergirum sp to decrease. It is very important to recognize the distribution and shifting trend of pathogenic fungi in the diagnosis, prevention and treatment of fungal keratitis. PMID- 15109457 TI - Expression of aFGF, bFGF, and FGFR1 in ovarian epithelial neoplasm. PMID- 15109458 TI - Combined transplantation of G-CSF primed allogeneic bone marrow cells and peripheral blood stem cells in treatment of severe aplastic anemia. PMID- 15109459 TI - Comparison of Epstein-Barr virus infection and 30 bp-deleted LMP1 gene among four histological types of nasopharyngeal carcinoma. PMID- 15109460 TI - Expression of survivin, a novel apoptosis inhibitor and cell cycle regulatory protein, in human gliomas. PMID- 15109461 TI - Role of Fas-FasL in insulitis in nonobese diabetic mouse. PMID- 15109462 TI - Toxin-coregulated pilus-loaded microparticles as a vaccine against Vibrio cholerae O139. PMID- 15109463 TI - Effect of FGF-BP on angiogenesis in squamous cell carcinoma. PMID- 15109464 TI - Changes in monocyte counts and expression of mCD(14) and HLA-DR in the peripheral blood of patients with severe acute respiratory syndrome. PMID- 15109465 TI - Association analysis of the cholecystokinin type A receptor gene in schizophrenia. PMID- 15109466 TI - Gene delivery to mice spermatogenic stem cells by Effectene. PMID- 15109467 TI - Composite valve graft combined with replacement of the ascending aorta and aortic arch in a patient with Marfan's syndrome. PMID- 15109468 TI - Primary aortoenteric fistula complicated by esophageal ulcer: case report. PMID- 15109469 TI - Systemic lupus erythematosus and thrombotic thrombocytopenic purpura: report of three cases. PMID- 15109470 TI - High rates of substance abuse among long-term travelers to the tropics: an interventional study. AB - BACKGROUND: Drug abuse constitutes a worldwide problem. The purpose of this study was to investigate the characteristics of illicit drug use during long-term travel to the tropics, and to estimate the effects of antidrug brochures provided before travel on the use of drugs during travel. METHODS: An interventional study was done on 1,000 travelers to the tropics aged 18 to 30 years. Pretravel brochures explaining the hazards of drug abuse were provided to 500 of them. A control group of 500 travelers did not receive the brochures. Questionnaires exploring drug abuse habits were sent to all travelers after their return. RESULTS: Among 223 travelers (108 and 115 of the intervention and control groups, respectively) who returned their questionnaires, 82 (36.8%, 95% CI 30.5-43.5%) had experienced illicit drug use during their trip, whereas only 52 of them had tried drugs before (p<0.01). More travelers had used drugs in the Far East (43.3%) than in South America (25.6%, p< 0.01). The strongest predictors of drug abuse were: the compound of female gender and travel to Asia (odds ratio (OR), 4.3), education 38.3 degrees C, and/or sore throat were included. Pilgrims with any other symptoms, especially myalgia and fatigue alone, were excluded, since many of the physical chores during the pilgrimage may contribute to such symptoms. RESULTS: Fifty-four patients (10.8%) had positive viral throat cultures. Of these, 27 (50%) were influenza B, 13 (24.1%) were HSV, 7 (12.9%) were RSV, 4 (7.4%) were parainfluenza, and 3 (5.6%) were influenza A. No enteroviruses or adenoviruses were detected, and no multiple infections were detected. Only 22 (4.7%) pilgrims received the influenza vaccine. When the results are applied to the total number of pilgrims in 2003, an estimate of 24,000 cases of influenza is obtained. CONCLUSION: The findings from this study suggest a high incidence of influenza as a cause of upper respiratory tract infection among pilgrims, estimated to be 24,000 cases per Hajj season, excluding those becoming ill from contact with Hajj pilgrims returning home. They also indicate a very low vaccination rate for the influenza vaccine; as well as poor knowledge of its existence. Continued surveillance during the Hajj pilgrimage is necessary. The influenza vaccine should be a priority for those attending the Hajj pilgrimage, and should also be considered for antiviral prophylaxis. PMID- 15109472 TI - Pneumonia among travelers returning from abroad. AB - BACKGROUND: Although respiratory tract infections represent a frequent cause of morbidity in travelers, and pneumonia a frequent cause of medical consultation among febrile travelers returning home, the etiologic spectrum of pneumonia in travelers has not been specifically studied. METHODS: We reviewed the medical charts of all travelers hospitalized during a 12-month period in our department with pneumonia after returning home. RESULTS: Seventeen patients (nine men, eight women, mean age 44 years, range 26 to 67 years) were included in this study. The etiology of pneumonia was established in 13 patients. Bacterial pneumonia was documented in 10 cases and was due to Streptococcus pneumoniae (n=2), Mycoplasma pneumoniae (n=2), Legionella pneumophila (n=1), Coxiella burnetti (n=1), Leptospira sp. (n=1) or Mycobacterium tuberculosis (n=3). Other etiologies included histoplasmosis, invasive schistosomiasis and dengue fever (one case each). CONCLUSION: These results show the wide range of causes of pneumonia among travelers returning from abroad. PMID- 15109473 TI - Do mosquito coils prevent malaria? A systematic review of trials. AB - BACKGROUND: Official guidelines commonly advise travelers to burn mosquito coils as one means of preventing malaria. The objective of this study was to discover if insecticide-containing mosquito coils (1) prevent mosquito bites and therefore malaria acquisition, and (2) are safe in terms of their adverse effects on human users. METHODS: We sought published and unpublished controlled trials in this area, by (1) contacting experts, (2) searching the Cochrane Library, (3) interrogating bibliographic databases, (4) Internet search, (5) citation scanning, (6) scanning conference proceedings, and (7) writing to manufacturers. RESULTS: Fifteen controlled trials of insecticide-containing mosquito coils met our predefined inclusion criteria. We found no controlled trials measuring the incidence of clinical malaria as an outcome. Studies tested the efficacy of coils in achieving mosquito bite reduction (reported in 14 studies), mosquito repellence (seven), deterrence (five), "knockdown" effect (five), and percentage mosquito mortality (seven). Of the 38 separate outcome measures reported, antimosquito efficacy was reported as positive for >95%. One trial reported no antimosquito effect at all, for one outcome only. Some insecticide classes and strengths were associated with better antimosquito outcomes than others. One trial identified possible adverse effects (irritation of the eyes and nose) in human users of this technology. CONCLUSIONS: There is no evidence that burning insecticide-containing mosquito coils prevents malaria acquisition. A randomized field trial should be conducted, with malaria incidence as a primary outcome. There is consistent evidence that burning coils inhibits nuisance biting by various mosquito species. The potential harmful effects of coil smoke on human users should be investigated. PMID- 15109474 TI - Status of malaria in the Kingdom of Bahrain: a 10-year review. AB - BACKGROUND: Malaria was eradicated in Bahrain about 20 years ago. However, because of a large immigrant population, cases of imported malaria continued to be seen. This paper presents an evaluation of the data from 1992 to 2001 to assess the levels of indigenous and imported malaria cases and the potential for reemergence of malaria transmission in the country. METHODS: Epidemiologic and parasitologic data on confirmed malaria cases during the review period were analyzed. Data on vector breeding activity were also analyzed to determine the potential for reemergence of local transmission. RESULTS: From 1992 to 2001, 1,572 cases of malaria were reported. All were imported malaria cases. There was a consistent decline in the number of cases, from 282 (the peak level) in 1992 to 54 in 2001. Eighty-four percent (1,318/1,572) of infections were contracted as a result of travel to or previous domicile in five countries, namely India, Pakistan, Sri Lanka, Bangladesh and Sudan. Plasmodium vivax was the etiologic agent in 85.6% (1,346/1,572) of cases, with Plasmodium falciparum accounting for 14% (220/1,572). During this period, the percentage of examined breeding sites where Anopheles mosquito larvae were identified remained fairly constant, ranging between 0.05% and 0.1%. CONCLUSIONS: From 1992 to 2001, there was a consistent decline in the number of imported malaria cases in Bahrain. There were no cases of locally acquired malaria during this period. The low level of vector breeding spots with Anopheles mosquito larvae suggests that the potential for reemergence of local transmission remains low. PMID- 15109475 TI - An estimate of the incidence of hepatitis A in unimmunized Canadian travelers to developing countries. AB - BACKGROUND: There is a paucity of data describing the risk of acquiring hepatitis A while traveling in the developing world. This paper uses available data to calculate the risk to Canadian travelers. METHODS: Information was gathered from Canadian and international sources on the following: the yearly incidence of hepatitis A among Canadians; the proportion of cases of hepatitis A associated with travel to developing countries; the number of days of such travel by Canadians per year; and the percentage of travelers immunized before departure. Calculations were performed on these figures to arrive at an estimated risk of infection for unimmunized Canadian travelers. RESULTS: The annual incidence of hepatitis A in Canada over the period 1996-2001, adjusted for underreporting, averaged 6.15 cases/100,000 people. During that time, Canadians traveled approximately 36.5 million days/year in developing countries. The literature shows that 4% to 28% (mean 16%) of cases are estimated to have been acquired abroad. It also shows that 14% to 24% (mean 19%) of such travelers are immunized before departure. Based on these figures, the risk of acquiring hepatitis A during 1 month of travel in the developing world is calculated to be approximately 1 case per 3,000 unimmunized travelers. CONCLUSION: Hepatitis A is an important travel-related disease, preventable by immunization. However, our calculations indicate that the risk of acquiring hepatitis A while traveling in the developing world is lower than some previously published estimates. The results represent an average for all types of travel to all such countries. The actual risk will vary considerably, depending on the destination and style of travel. PMID- 15109476 TI - Impact of severe acute respiratory syndrome (SARS) on travel and population mobility: implications for travel medicine practitioners. PMID- 15109477 TI - Dengue fever and long thoracic nerve palsy in a traveler returning from Thailand. PMID- 15109478 TI - Plasmodium and Borrelia co-infection. PMID- 15109479 TI - From travelers' diarrhea to abdominal surgery: report of three cases. PMID- 15109480 TI - Excellent booster response 4 to 8 years after a single primary dose of an inactivated hepatitis A vaccine. PMID- 15109481 TI - Zebra bite to a South African tourist. PMID- 15109482 TI - Is quartan malaria safely prevented by mefloquine prophylaxis? PMID- 15109483 TI - Global impact of severe acute respiratory syndrome: measures to prevent importation into Saudi Arabia. PMID- 15109484 TI - A case of in-flight transmission of severe acute respiratory syndrome (SARS): SARS serology positive. PMID- 15109485 TI - Hepatitis A+B vaccine in elderly persons. PMID- 15109486 TI - Selected bibliogaphy. PMID- 15109487 TI - Telomerase regulation at the telomere: a binary switch. AB - Telomerase is known to preferentially elongate the shortest telomeres in a cell. Using an elegant yeast assay, Texeira et al. (2004 [this issue of Cell]) address what aspect of telomerase action is regulated by telomere length: the frequency or the extent of telomere elongation. They show that short telomeres are elongated more frequently than long telomeres, arguing that telomeres switch between two states, one that allows telomere extension and one that does not. PMID- 15109488 TI - Hitchhiking without covalent integration. AB - In eukaryotes, many latent viruses attach to mitotic chromosomes noncovalently for effective partitioning in dividing cells. For different viruses, the cis and trans elements encoded by the episomes have been effectively defined but the chromosomal "receptors" for such tethering have remained elusive. In this issue of Cell, give us a first insight into the cellular protein machinery important for animal papillomavirus retention. PMID- 15109489 TI - The spindle gets bigger. AB - Although cortical nonmuscle myosin II has long been implicated in cytokinetic aspects of cell division, there has never been strong evidence that it plays a role in the organization of the mitotic spindle. ([this issue of Cell]) use a number of methods to show that cortical myosin II is in fact important for spindle assembly in higher eukaryotic cells, specifically for the complete separation of centrosomes after nuclear envelope breakdown. PMID- 15109490 TI - Balancing acts: molecular control of mammalian iron metabolism. AB - Iron is ubiquitous in the environment and in biology. The study of iron biology focuses on physiology and homeostasis-understanding how cells and organisms regulate their iron content, how diverse tissues orchestrate iron allocation, and how dysregulated iron homeostasis leads to common hematological, metabolic, and neurodegenerative diseases. This has provided novel insights into gene regulation and unveiled remarkable links to the immune system. PMID- 15109491 TI - Structural basis for transcription regulation by alarmone ppGpp. AB - Guanosine-tetraphosphate (ppGpp) is a major regulator of stringent control, an adaptive response of bacteria to amino acid starvation. The 2.7 A resolution structure of the Thermus thermophilus RNA polymerase (RNAP) holoenzyme in complex with ppGpp reveals that ppGpp binds to the same site near the active center in both independent RNAP molecules in the crystal but in strikingly distinct orientations. Binding is symmetrical with respect to the two diphosphates of ppGpp and is relaxed with respect to the orientation of the nucleotide base. Different modes of ppGpp binding are coupled with asymmetry of the active site configurations. The results suggest that base pairing of ppGpp with cytosines in the nontemplate DNA strand might be an essential component of transcription control by ppGpp. We present experimental evidence highlighting the importance of base-specific contacts between ppGpp and specific cytosine residues during both transcription initiation and elongation. PMID- 15109492 TI - Identification of a human endonuclease complex reveals a link between tRNA splicing and pre-mRNA 3' end formation. AB - tRNA splicing is a fundamental process required for cell growth and division. The first step in tRNA splicing is the removal of introns catalyzed in yeast by the tRNA splicing endonuclease. The enzyme responsible for intron removal in mammalian cells is unknown. We present the identification and characterization of the human tRNA splicing endonuclease. This enzyme consists of HsSen2, HsSen34, HsSen15, and HsSen54, homologs of the yeast tRNA endonuclease subunits. Additionally, we identified an alternatively spliced isoform of SEN2 that is part of a complex with unique RNA endonuclease activity. Surprisingly, both human endonuclease complexes are associated with pre-mRNA 3' end processing factors. Furthermore, siRNA-mediated depletion of SEN2 exhibited defects in maturation of both pre-tRNA and pre-mRNA. These findings demonstrate a link between pre-tRNA splicing and pre-mRNA 3' end formation, suggesting that the endonuclease subunits function in multiple RNA-processing events. PMID- 15109493 TI - Telomere length homeostasis is achieved via a switch between telomerase- extendible and -nonextendible states. AB - Telomerase counteracts telomere erosion that stems from incomplete chromosome end replication and nucleolytic processing. A precise understanding of telomere length homeostasis has been hampered by the lack of assays that delineate the nonuniform telomere extension events of single chromosome molecules. Here, we measure telomere elongation at nucleotide resolution in Saccharomyces cerevisiae. The number of nucleotides added to a telomere in a single cell cycle varies between a few to more than 100 nucleotides and is independent of telomere length. Telomerase does not act on every telomere in each cell cycle, however. Instead, it exhibits an increasing preference for telomeres as their lengths decline. Deletion of the telomeric proteins Rif1 or Rif2 gives rise to longer telomeres by increasing the frequency of elongation events. Thus, by taking a molecular snapshot of a single round of telomere replication, we demonstrate that telomere length homeostasis is achieved via a switch between telomerase-extendible and nonextendible states. PMID- 15109494 TI - Telomere maintenance requires the RAD51D recombination/repair protein. AB - The five RAD51 paralogs (RAD51B, RAD51C, RAD51D, XRCC2, and XRCC3) are required in mammalian cells for normal levels of genetic recombination and resistance to DNA-damaging agents. We report here that RAD51D is also involved in telomere maintenance. Using immunofluorescence labeling, electron microscopy, and chromatin immunoprecipitation assays, RAD51D was shown to localize to the telomeres of both meiotic and somatic cells. Telomerase-positive Rad51d(-/-) Trp53(-/-) primary mouse embryonic fibroblasts (MEFs) exhibited telomeric DNA repeat shortening compared to Trp53(-/-) or wild-type MEFs. Moreover, elevated levels of chromosomal aberrations were detected, including telomeric end-to-end fusions, a signature of telomere dysfunction. Inhibition of RAD51D synthesis in telomerase-negative immortalized human cells by siRNA also resulted in telomere erosion and chromosome fusion. We conclude that RAD51D plays a dual cellular role in both the repair of DNA double-strand breaks and telomere protection against attrition and fusion. PMID- 15109495 TI - Interaction of the bovine papillomavirus E2 protein with Brd4 tethers the viral DNA to host mitotic chromosomes. AB - The papillomavirus E2 protein tethers viral genomes to host mitotic chromosomes to ensure genome maintenance. We have identified the bromodomain protein Brd4 as a major cellular interacting partner of the bovine papillomavirus E2. Brd4 associates with mitotic chromosomes and colocalizes with E2 on mitotic chromosomes. The site of E2 binding maps to the C-terminal domain of Brd4. Expression of this C-terminal Brd4 domain functions in a dominant-negative manner to abrogate the colocalization of E2 with Brd4 on mitotic chromosomes, to block association of the viral episomes with Brd4, and to inhibit BPV-1 DNA-mediated cellular transformation. Brd4 also associates with HPV16 E2, indicating that Brd4 binding may be a shared property of all papillomavirus E2 proteins. The interaction of E2 with Brd4 is required to ensure the tethering of viral genomes to the host mitotic chromosomes for persistence of viral episomes in PV-infected cells. PMID- 15109496 TI - Myosin II-dependent cortical movement is required for centrosome separation and positioning during mitotic spindle assembly. AB - The role of myosin II in mitosis is generally thought to be restricted to cytokinesis. We present surprising new evidence that cortical myosin II is also required for spindle assembly in cells. Drug- or RNAi-mediated disruption of myosin II in cells interferes with normal spindle assembly and positioning. Time lapse movies reveal that these treatments block the separation and positioning of duplicated centrosomes after nuclear envelope breakdown (NEBD), thereby preventing the migration of the microtubule asters to opposite sides of chromosomes. Immobilization of cortical movement with tetravalent lectins produces similar spindle defects to myosin II disruption and suggests that myosin II activity is required within the cortex. Latex beads bound to the cell surface move in a myosin II-dependent manner in the direction of the separating asters. We propose that after NEBD, completion of centrosome separation and positioning around chromosomes depends on astral microtubule connections to a moving cell cortex. PMID- 15109497 TI - Nkx2-5 pathways and congenital heart disease; loss of ventricular myocyte lineage specification leads to progressive cardiomyopathy and complete heart block. AB - Human mutations in Nkx2-5 lead to progressive cardiomyopathy and conduction defects via unknown mechanisms. To define these pathways, we generated mice with a ventricular-restricted knockout of Nkx2-5, which display no structural defects but have progressive complete heart block, and massive trabecular muscle overgrowth found in some patients with Nkx2-5 mutations. At birth, mutant mice display a hypoplastic atrioventricular (AV) node and then develop selective dropout of these conduction cells. Transcriptional profiling uncovered the aberrant expression of a unique panel of atrial and conduction system-restricted target genes, as well as the ectopic, high level BMP-10 expression in the adult ventricular myocardium. Further, BMP-10 is shown to be necessary and sufficient for a major component of the ventricular muscle defects. Accordingly, loss of ventricular muscle cell lineage specification into trabecular and conduction system myocytes is a new mechanistic pathway for progressive cardiomyopathy and conduction defects in congenital heart disease. PMID- 15109498 TI - ATF4 is a substrate of RSK2 and an essential regulator of osteoblast biology; implication for Coffin-Lowry Syndrome. AB - Coffin-Lowry Syndrome (CLS) is an X-linked mental retardation condition associated with skeletal abnormalities. The gene mutated in CLS, RSK2, encodes a growth factor-regulated kinase. However, the cellular and molecular bases of the skeletal abnormalities associated with CLS remain unknown. Here, we show that RSK2 is required for osteoblast differentiation and function. We identify the transcription factor ATF4 as a critical substrate of RSK2 that is required for the timely onset of osteoblast differentiation, for terminal differentiation of osteoblasts, and for osteoblast-specific gene expression. Additionally, RSK2 and ATF4 posttranscriptionally regulate the synthesis of Type I collagen, the main constituent of the bone matrix. Accordingly, Atf4-deficiency results in delayed bone formation during embryonic development and low bone mass throughout postnatal life. These findings identify ATF4 as a critical regulator of osteoblast differentiation and function, and indicate that lack of ATF4 phosphorylation by RSK2 may contribute to the skeletal phenotype of CLS. PMID- 15109500 TI - Is intranasal ketamine an appropriate treatment for chronic non-cancer breakthrough pain? PMID- 15109499 TI - Foxo transcription factors induce the atrophy-related ubiquitin ligase atrogin-1 and cause skeletal muscle atrophy. AB - Skeletal muscle atrophy is a debilitating response to fasting, disuse, cancer, and other systemic diseases. In atrophying muscles, the ubiquitin ligase, atrogin 1 (MAFbx), is dramatically induced, and this response is necessary for rapid atrophy. Here, we show that in cultured myotubes undergoing atrophy, the activity of the PI3K/AKT pathway decreases, leading to activation of Foxo transcription factors and atrogin-1 induction. IGF-1 treatment or AKT overexpression inhibits Foxo and atrogin-1 expression. Moreover, constitutively active Foxo3 acts on the atrogin-1 promoter to cause atrogin-1 transcription and dramatic atrophy of myotubes and muscle fibers. When Foxo activation is blocked by a dominant negative construct in myotubes or by RNAi in mouse muscles in vivo, atrogin-1 induction during starvation and atrophy of myotubes induced by glucocorticoids are prevented. Thus, forkhead factor(s) play a critical role in the development of muscle atrophy, and inhibition of Foxo factors is an attractive approach to combat muscle wasting. PMID- 15109501 TI - Experimental approach to CRPS. PMID- 15109502 TI - Lowering fear-avoidance and enhancing function through exposure in vivo. A multiple baseline study across six patients with back pain. AB - This study investigated the effects of an exposure in vivo treatment for chronic pain patients with high levels of fear and avoidance. The fear-avoidance model offers an enticing explanation of why some back pain patients develop persistent disability, stressing the role of catastrophic interpretations; largely fueled by beliefs and expectations that activity will cause injury and will worsen the pain problem. Recently, an exposure in vivo treatment was developed that aims to enhance function by directly addressing these fears and expectations. The purpose of this study was to describe the short-term, consequent effect of an exposure in vivo treatment. The study employed a multiple baseline design with six patients who were selected based on their high levels of fear and avoidance. The results demonstrated clear decreases in rated fear and avoidance beliefs while function increased substantially. These improvements were observed even though rated pain intensity actually decreased somewhat. Thus, the results replicate and extend the findings of previous studies to a new setting, with other therapists and a new research design. These results, together with the initial studies, provide a basis for pursuing and further developing the exposure technique and to test it in group designs with larger samples. PMID- 15109503 TI - Safety and efficacy of intranasal ketamine for the treatment of breakthrough pain in patients with chronic pain: a randomized, double-blind, placebo-controlled, crossover study. AB - Few placebo-controlled trials have investigated the treatment of breakthrough pain (BTP) in patients with chronic pain. We evaluated the efficacy and safety of intranasal ketamine for BTP in a randomized, double-blind, placebo-controlled, crossover trial. Twenty patients with chronic pain and at least two spontaneous BTP episodes daily self-administered up to five doses of intranasal ketamine or placebo at the onset of a spontaneous BTP episode (pain intensity > or =5 on a 0 10 scale). Two BTP episodes at least 48 h apart were treated with either ketamine or placebo. Patients reported significantly lower BTP intensity following intranasal ketamine than after placebo (P < 0.0001) with pain relief within 10 min of dosing and lasting for up to 60 min. No patient in the ketamine group required his/her usual rescue medication to treat the BTP episode, while seven out of 20 (35%) patients in placebo group did (P = 0.0135). Intranasal ketamine was well tolerated with no serious adverse events. After ketamine administration, four patients reported a transient change in taste, one patient reported rhinorrhea, one patient reported nasal passage irritation, and two patients experienced transient elevation in blood pressure. A side effect questionnaire administered 60 min and 24 h after drug or placebo administration elicited no reports of auditory or visual hallucinations. These data suggest that intranasal administration of ketamine provides rapid, safe and effective relief for BTP. PMID- 15109504 TI - Involvement of GABAergic modulation of the nucleus submedius (Sm) morphine induced antinociception. AB - Previous studies have shown that microinjection of morphine into the nucleus submedius (Sm) of the thalamus produces antinociception. The aim of the current study was to examine whether gamma-aminobutyric acid (GABA)ergic terminals in the Sm were involved in this antinociception. Under light anesthesia, the GABA(A) receptor antagonist bicuculline or agonist muscimol was microinjected into the Sm of the thalamus in Sm non-morphine-treated (control) or Sm morphine-treated (microinjection into the Sm in the thalamus) rats. Tail flick latencies (TFL) were measured in each of these groups of rats every 5 min. Bicuculline (100, 200, 500 ng in 0.5 microL) depressed the TF reflex in a dose-dependent fashion, and this effect was blocked by microinjection of the opioid receptor antagonist naloxone (0.5 microg) into the same Sm site. A small dose (100 ng) of bicuculline microinjected into Sm significantly enhanced the morphine-evoked inhibition of TF reflex. In contrast, administration of muscimol (250 ng) did not significantly influence the TF reflex in Sm non-morphine-treated rats, but it significantly attenuated the morphine-induced antinociception in the Sm morphine-treated rats. These results suggest that locally released GABA acting at GABA(A) receptors is involved in the modulation of Sm morphine-induced antinociception, and support the hypothesis that a disinhibitory effect elicited by morphine on GABAergic terminals in Sm may lead to activation of the Sm-ventrolateral orbital cortex (VLO)-perioqueductal gray (PAG) brainstem descending inhibitory system and depression of the nociceptive inputs at the spinal cord level. PMID- 15109505 TI - Pain reducing effect of three types of transcutaneous electrical nerve stimulation in patients with chronic pain: a randomized crossover trial. AB - Transcutaneous electrical nerve stimulation (TENS) is a frequently applied therapy in chronic pain although evidence for effectiveness is inconclusive. Several types of TENS, based on different combinations of frequency, pulse duration and intensity, exist. The precise mechanism of action and the relevance of combinations of stimulus parameters are still unclear. To compare the effectiveness of three types of TENS we conducted a randomized, single blinded crossover trial. Patients received two times a 2-week period of daily TENS treatment, separated by a washout period of 2 weeks. In total, 180 chronic pain patients were randomized into three groups. In group 1, high frequency, low intensity TENS (HFT) was compared with high frequency, high intensity TENS (HIT). In groups 2 and 3, HFT and HIT were compared with a control TENS (COT). The order of applying the different modalities of TENS in each group was also randomized. Primary outcome was the patient's overall assessment of effectiveness and pain reduction (VAS). No differences were found in patient's assessment or pain reducing effect between the three groups, indicating no superiority of one type of TENS. In total, 56% continued TENS after the 2-week treatment period. At 6 months, 42% of all patients still used TENS. We concluded that there were no differences in effectiveness for the three types of TENS used in this study. Because no placebo group was included, no definite conclusions on effectiveness of TENS in general in the treatment of chronic pain could be made. PMID- 15109506 TI - Effects of intensity and locus of painful stimulation on postural stability. AB - Stimulation of small diameter afferents can influence motor behavior. Little is known about how a prolonged painful stimulation of these small afferents may affect essential motor behavior such as the maintenance of an erect stance. The present study documents the effects of 10-s weak, moderate and extreme painful stimulations applied to the dorsum of the feet on the postural stability. Also, the moderate painful stimulation was applied to the metacarpal heads to determine if a painful stimulation to a limb not involved in the maintenance of the erect stance affects the postural control mechanisms. Increasing the intensity of the painful stimulation applied to the feet yielded larger postural oscillations whereas stimulation to the hands did not affect the control of posture. This suggests that the painful stimulation mainly affected the postural control mechanisms via sensorimotor processes rather than via cognitive resources related to the perception of pain. PMID- 15109507 TI - Topical clonazepam in stomatodynia: a randomised placebo-controlled study. AB - Stomatodynia is characterised by a spontaneous burning pain in the oral mucosa without known cause or recognised treatment. The purpose of this double-blind, randomised, multicentre parallel group study was to evaluate the efficacy of the topical use of clonazepam. Forty-eight patients (4 men and 44 women, aged 65+/ 2.1 years) were included, of whom 41 completed the study. The patients were instructed to suck a tablet of 1 mg of either clonazepam or placebo and hold their saliva near the pain sites in the mouth without swallowing for 3 min and then to spit. This protocol was repeated three times a day for 14 days. The intensity was evaluated by a 11-point numerical scale before the first administration and then after 14 days. Two weeks after the beginning of treatment, the decrease in pain scores was 2.4+/-0.6 and 0.6+/-0.4 in the clonazepam and placebo group, respectively (P = 0.014). Similar effects were obtained in an intent-to-treat analysis (P = 0.027). The blood concentration of clonazepam was similar whether it was measured 14 days after sucking a tablet three times a day or during the 5 h that followed sucking a single tablet (n = 5). It is hypothesised that clonazepam acts locally to disrupt the mechanism(s) underlying stomatodynia. PMID- 15109508 TI - A quantitative somatosensory testing of pain threshold in individuals with mental retardation. AB - The commonly held view, mainly based on behavioral observations, is that individuals with mental retardation (MR) have a decreased sensitivity to pain. However, the sensitivity to noxious stimuli was not systematically measured in these individuals. For this purpose we developed an experimental protocol with which we trained individuals with mild MR (unspecified MR and Down's syndrome) in heat-pain threshold (HPT) measurement on the hand, and then performed the measurement using both the method of limits (MLI) which relies on reaction time (RT) and the method of levels (MLE) which is RT-free. This allowed for an indirect assessment of the RT and conduction velocity (CV) of these individuals. We found that HPT in individuals with unspecified MR (41.23+/-1.86 degrees C) and Down's syndrome (40.96+/-2.93 degrees C) was significantly lower than that of controls (42.86+/-2.42 degrees C) when measured with the MLE (P < 0.05). With the MLI no significant differences in HPT were found between the groups. However, the RT and CV values of individuals with unspecified MR and Down's syndrome were significantly lower compared to controls (e.g. mean RT of 1.86 and 2.55 compared to 1.2 s, respectively, P < 0.01). From this work it would appear that individuals with MR are not only pain-sensitive, but also more sensitive to heat pain than normal. It is suggested that computerized quantitative testing of pain threshold is feasible in individuals with MR preferably by using RT-free methods (e.g. the MLE) due to the low RT and CV values exhibited by them. PMID- 15109509 TI - Endogenous peripheral antinociception in early inflammation is not limited by the number of opioid-containing leukocytes but by opioid receptor expression. AB - Endogenous inhibition of inflammatory pain is mediated by leukocytes that secrete opioid peptides upon exposure to stress (cold water swim stress, CWS) or after local injection of corticotropin releasing factor (CRF). Since in early inflammation few opioid-containing leukocytes are detected and since peripheral opioid-mediated antinociception is low we examined whether antinociception could be augmented by increased recruitment of opioid-containing polymorphonuclear cells (PMN). Rats were intraplantarly (i.pl.) injected with Freund's complete adjuvant (FCA) and with the PMN-recruiting chemokine macrophage inflammatory protein-2 (MIP-2, 1-10 microg; control: saline) for 2 h. Intraplantar leukocytes were quantified by flow cytometry. Paw pressure threshold (PPT) was determined before and after exposure to CWS, i.pl. injection of CRF and opioid peptides. Opioid receptors (OR) were measured by binding studies in dorsal root ganglia (DRG) and by immunohistochemistry in the paw. Our studies showed that (i) MIP-2 injection dose-dependently augmented recruitment of PMN and opioid-containing leukocytes (5-fold increase in cells/paw, P < 0.05), (ii) PPT was not different between groups at baseline and after CWS or CRF (maximum MPE: 20+/-2.3-29+/-7.2%, P < 0.05), (iii) injection of opioid peptides dose-dependently increased the PPT (P < 0.05, maximum MPE: and 18+/-2.6-21+/-3.6%), (iv) MOR (micro OR, MOP) binding sites in the ipsilateral DRG were unchanged (24+/-2-22+/-1.2 fmol/mg protein, P < 0.05, ANOVA) and (v) the number of MOR and DOR (delta OR, DOP) stained nerve fibers in peripheral tissue were unaltered (both P > 0.05, t-test). In summary, antinociception during early inflammation is apparently not limited by the number of opioid-containing leukocytes but by OR availability. PMID- 15109510 TI - Electrophysiological characterisations of rat lamina I dorsal horn neurones and the involvement of excitatory amino acid receptors. AB - Lamina I of the spinal cord plays a key role in sensory transmission between afferent activity and the CNS. Studies have shown lamina I neurones to have distinct response properties compared to deep dorsal horn neurones, but little is known regarding excitatory amino acid mechanisms in their responses. Spinal electrophysiological recordings of lamina I neurones confirmed that the majority of these neurones (74%) are nociceptive specific (NS) in their responses, of which 18% can be termed polymodal nociceptive (HPC) (13% of the total population). The remainder (26%) were wide dynamic range. Lamina I neurones had smaller mechanical and heat-evoked responses compared to deeper dorsal horn neurones. The electrically evoked responses were also smaller, with a distinct lack of an NMDA-mediated 'wind-up' effect. NBQX (AMPA receptor antagonist, 0.5, 5, 50 microg/50 microl) produced dose-dependent inhibitions of the electrically evoked neuronal responses, but APV (NMDA receptor antagonist, 50, 100, 500 microg/50 microl) had minimal effects on their responses. These results implicate mainly AMPA receptors in the responses of lamina I neurones. Bicuculline (GABA(A) receptor antagonist, 0.5, 5, 50 microg/50 microl) demonstrated a role exerted by GABA(A) receptors in the control of A-delta fibre-mediated mechanical responses in lamina I. Overall, this study describes a high threshold, AMPA receptor possessing population of lamina I neurones, which seem to lack functional NMDA receptors, and are partially controlled by GABA(A) receptor activity. PMID- 15109511 TI - Psychological factors in pelvic/urogenital pain: the influence of site of pain versus sex. AB - Chronic pelvic pain (CPP), a fairly common gynecological complaint in women, has been associated with multiple psychological sequelae, including depression and somatization. Previous work has compared these patients to gynecological controls and women with headache, but has failed to include male comparison groups with a comparable site of chronic pain. In order to test possible sex and pain site differences, the present study compared 22 women with CPP, 22 men with either penile or testicular pain, 22 women with low back pain and 28 men with low back pain referred for a psychological evaluation as part of multidisciplinary pain treatment. Depression, coping, pain intensity and interference were assessed. Two way analyses of variance (sex by pain site) were conducted to determine if there were group differences on demographic variables and medical history. Pain duration, age, and pain severity differed among the groups and were entered as covariates in hierarchical regression analyses designed to identify predictors of adjustment and pain coping. Sex and pain site did not contribute independently to the prediction of depressive symptoms. Pain site predicted physical functioning with low back pain patients reporting greater pain-related interference. Similar findings were demonstrated for coping. A variety of pain-coping strategies, including catastrophizing, were more frequently utilized by low back pain patients, regardless of sex. In the present study, pain severity and pain site explained more variance in depressive symptoms, physical functioning, and pain coping than sex. PMID- 15109512 TI - Substance P signaling contributes to the vascular and nociceptive abnormalities observed in a tibial fracture rat model of complex regional pain syndrome type I. AB - Wrist and ankle fractures are the most frequent causes of complex regional pain syndrome (CRPS type I). The current study examined the temporal development of vascular, nociceptive and bony changes after distal tibial fracture in rats and compared these changes to those observed after cast immobilization in intact normal rats. After baseline testing the right distal tibial was fractured and the hindlimb casted. A control group was simply casted without fracturing the tibia. After 4 weeks the casts were removed and the rats retested. Subsequent testing was performed at 6, 8, 10, 16, and 20 weeks after onset of treatment. Distal tibial fracture or cast immobilization alone generated chronic hindlimb warmth, edema, spontaneous protein extravasation, allodynia, and periarticular osteoporosis, changes resembling those observed in CRPS. Hindlimb warmth and allodynia resolved much more quickly after cast immobilization than after fracture. Previously we observed that the substance P receptor (NK(1)) antagonist LY303870 reversed vascular and nociceptive changes in a sciatic section rat model of CRPS type II. Postulating that facilitated substance P signaling may also contribute to the vascular and nociceptive abnormalities observed after tibial fracture or cast immobilization, we attempted to reverse these changes with LY303870. Hindpaw warmth, spontaneous extravasation, edema, and allodynia were inhibited by LY303870. Collectively, these data support the hypotheses that the distal tibial fracture model simulates CRPS, immobilization alone can generate a syndrome resembling CRPS, and substance P signaling contributes to the vascular and nociceptive changes observed in these models. PMID- 15109513 TI - Analgesic effect of amitriptyline in chronic tension-type headache is not directly related to serotonin reuptake inhibition. AB - The tricyclic antidepressant amitriptyline is the only documented and most widely used prophylactic drug for chronic tension-type headache (CTTH). However, it is not fully clarified whether the serotonin (5-HT) reuptake inhibition plays a major role for the analgesic effect of amitriptyline. To explore the importance of 5-HT reuptake inhibition for mechanism of action of the analgesic effect of amitriptyline we investigated platelet 5-HT levels during preventive treatment of CTTH with amitriptyline, the selective serotonin reuptake inhibitor citalopram, and placebo. Thirty-four patients with CTTH were given preventive treatment with amitriptyline 75 mg/day, the selective 5-HT reuptake inhibitor citalopram 20 mg/day, and placebo in a 32-week, double-blind, placebo-controlled, three-way crossover trial. Blood samples were collected in the last week of each treatment period. Platelet 5-HT was used as a measure of 5-HT reuptake inhibition and determined by high performance liquid chromatography. Area under the headache curve was 308 (157-715) (median with quartiles in parentheses) with amitriptyline and significantly lower than 377 (158-1121) with citalopram (P = 0.04) and 441 (178-1408) with placebo (P = 0.002). There was no difference between citalopram and placebo (P = 0.23). Platelet 5-HT was 0.4 (0.3-0.7) x 10(-18)mol/platelet with citalopram, which was significantly lower than 1.7 (1.2-2.4) x 10( 18)mol/platelet with amitriptyline (P < 0.001), and 3.5 (2.8-4.3) x 10( 18)mol/platelet with placebo (P < 0.001). The lower platelet 5-HT during treatment with citalopram than amitriptyline indicates that 5-HT reuptake was most effectively inhibited by citalopram. In contrast, amitriptyline was most effective in reduction of headache. This suggests that the analgesic effect of amitriptyline in CTTH is not solely due to 5-HT reuptake inhibition and that other mechanisms such as norepinephrine reuptake inhibition, NMDA receptor antagonism, blockade of muscarinic receptors and ion channels should be addressed in the future research. PMID- 15109514 TI - Health status as measured by SF-36 reflects changes and predicts outcome in chronic musculoskeletal pain: a 3-year follow up study in the general population. AB - The SF-36 is a well-validated health status instrument measuring eight different health concepts. One aim of this study was to compare health status as measured by SF-36 in subjects from the general population with no chronic pain (NCP), chronic regional pain (CRP), and chronic widespread pain (CWP). A second aim was to assess if SF-36 could reflect changes in pain status over time. A third aim was to study if health status at baseline, measured by SF-36, could predict pain status 3 years later. The study was designed as a 3-year follow up with a postal questionnaire, including the SF-36 health survey, to 2357 subjects from the general population aged 20-74 years. The results were controlled for age, sex, co morbidity, and socio-economic status. At baseline, all eight health concepts of SF-36 discriminated between subgroups with NCP, CRP and CWP. Changes in SF-36 over the 3-year follow up time coincided with improvement or deterioration of pain status. Baseline SF-36 scores predicted pain outcome 3 years later. These results support that both physical and mental aspects of health status as measured by SF-36 are affected by the burden of musculoskeletal pain, are sensitive to changes in pain status, and also predict the further development of pain. PMID- 15109515 TI - The effect of multiple stimuli on the modulation of the 'nociceptive' blink reflex. AB - The 'nociceptive' blink reflex is a method of examining human trigeminal pain pathways. We explored temporal summation of this reflex by using a train of pulses, rather than a single pulse, and remote activation of diffuse noxious inhibitory control (DNIC), to improve reliability, flexibility and nociceptive specificity of this technique. The R2 component of the nociceptive blink reflex response (nR2) was assessed in 28 healthy volunteers using between 1 and 7 pulses per stimulus train (inter-pulse interval 5 ms). The effect of DNIC on single-, double-, and triple-pulse nR2 was investigated. Compared to single pulses, double and triple pulses increased the sensation of pain, reduced the tactile and pain thresholds, and facilitated the blink reflex responses (reduced onset latency, increased magnitude and persistence of nR2). The maximal reflex facilitation was achieved using a triple pulse. Higher pulse numbers had no additional facilitatory effect. Activation of the DNIC system using heterotopic pain suppressed the nR2 evoked by double and triple stimulation by 16 and 42%, respectively, but not the nR2 from a single pulse. Stimulation with double and triple pulses may be more suitable to study influences on nociceptive pathways than single pulses and may widen the methodological flexibility of the nociceptive blink reflex technique. This technique may be useful in studying the trigeminal nociceptive system with particular reference to primary headache disorders and their neuropharmacology. PMID- 15109516 TI - Chronic pain patients are impaired on an emotional decision-making task. AB - Chronic pain can result in anxiety, depression and reduced quality of life. However, its effects on cognitive abilities have remained unclear although many studies attempted to psychologically profile chronic pain. We hypothesized that performance on an emotional decision-making task may be impaired in chronic pain since human brain imaging studies show that brain regions critical for this ability are also involved in chronic pain. Chronic back pain (CBP) patients, chronic complex regional pain syndrome (CRPS) patients, and normal volunteers (matched for age, sex, and education) were studied on the Iowa Gambling Task, a card game developed to study emotional decision-making. Outcomes on the gambling task were contrasted to performance on other cognitive tasks. The net number of choices made from advantageous decks after subtracting choices made from disadvantageous decks on average was 22.6 in normal subjects (n = 26), 13.4 in CBP patients (n = 26), and -9.5 in CRPS patients (n = 12), indicating poor performance in the patient groups as compared to the normal controls (P < 0.004). Only pain intensity assessed during the gambling task was correlated with task outcome and only in CBP patients (r = -0.75, P < 0.003). Other cognitive abilities, such as attention, short-term memory, and general intelligence tested normal in the chronic pain patients. Our evidence indicates that chronic pain is associated with a specific cognitive deficit, which may impact everyday behavior especially in risky, emotionally laden, situations. PMID- 15109517 TI - Spinal cord stimulation for patients with failed back surgery syndrome or complex regional pain syndrome: a systematic review of effectiveness and complications. AB - We conducted a systematic review of the literature on the effectiveness of spinal cord stimulation (SCS) in relieving pain and improving functioning for patients with failed back surgery syndrome and complex regional pain syndrome (CRPS). We also reviewed SCS complications. Literature searches yielded 583 articles, of which seven met the inclusion criteria for the review of SCS effectiveness, and 15 others met the criteria only for the review of SCS complications. Two authors independently extracted data from each article, and then resolved discrepancies by discussion. We identified only one randomized trial, which found that physical therapy (PT) plus SCS, compared with PT alone, had a statistically significant but clinically modest effect at 6 and 12 months in relieving pain among patients with CRPS. Similarly, six other studies of much lower methodological quality suggest mild to moderate improvement in pain with SCS. Pain relief with SCS appears to decrease over time. The one randomized trial suggested no benefits of SCS in improving patient functioning. Although life-threatening complications with SCS are rare, other adverse events are frequent. On average, 34% of patients who received a stimulator had an adverse occurrence. We conclude with suggestions for methodologically stronger studies to provide more definitive data regarding the effectiveness of SCS in relieving pain and improving functioning, short- and long-term, among patients with chronic pain syndromes. PMID- 15109518 TI - The cyclooxygenase isozyme inhibitors parecoxib and paracetamol reduce central hyperalgesia in humans. AB - Non-steroidal antiinflammatory drugs (NSAIDs) are known to induce analgesia mainly via inhibition of cyclooxygenase (COX). Although the inhibition of COX in the periphery is commonly accepted as the primary mechanism, experimental and clinical data suggest a potential role for spinal COX-inhibition to produce antinociception and reduce hypersensitivity. We used an experimental model of electrically evoked pain and hyperalgesia in human skin to determine the time course of central analgesic and antihyperalgesic effects of intravenous parecoxib and paracetamol (acetaminophen). Fourteen subjects were enrolled in this randomized, double blind, and placebo controlled cross-over study. In three sessions, separated by 2-week wash-out periods, the subjects received intravenous infusions of 40 mg parecoxib, 1000 mg paracetamol, or placebo. The magnitude of pain and areas of pinprick-hyperalgesia and touch evoked allodynia were repeatedly assessed before, and for 150 min after the infusion. While pain ratings were not affected, parecoxib as well as paracetamol significantly reduced the areas of secondary hyperalgesia to pinprick and touch. In conclusion, our results provide clear experimental evidence for the existence of central antihyperalgesia induced by intravenous infusion of two COX inhibitors, parecoxib and paracetamol. Since the electrical current directly stimulated the axons, peripheral effects of the COX inhibitors on nociceptive nerve endings cannot account for the reduction of hyperalgesia. Thus, besides its well-known effects on inflamed peripheral tissues, inhibition of central COX provides an important mechanism of NSAID-mediated antihyperalgesia in humans. PMID- 15109519 TI - Development of and recovery from long-term pain. A 6-year follow-up study of a cross-section of the adult Danish population. AB - A 6-year follow-up study of a cross-section of the adult Danish population, based on data from the Danish Health and Morbidity Surveys in 1994 and 2000 is presented. The pain populations were identified through the pain intensity verbal rating scale (VRS) included in the Short Form 36. The 2000 survey also included a question on duration of pain (>6 months). Using this as the 'gold standard', a validation study was performed, which identified the highest accuracy (85%) at the VRS cut-off level: no pain, very mild, or mild pain (control group) versus moderate, severe, or very severe pain (pain group). The cohort comprised 2649 individuals, representative of the Danish population. Prevalence rates of pain in 1994 and 2000 were 13.5 and 15.7%, respectively. The cumulated 6-year incidence of pain development was 10.7%, and the cumulated incidence of pain recovery was 52.1%. During the investigated period, 9.2% of individuals moved from a 'no pain status' to a 'pain status', 7% moved from a 'pain status' to a 'no pain status', and 6.5% maintained their 'pain status'. Significant risk factors for pain development were female gender [odds ratio (OR) 1.5, 95% confidence interval (CI) 1.2-2.0], short education (OR 1.5, CI 1.0-2.2), poor self-rated health (OR 3.3, CI 2.4-4.7), and having at least one long-standing disease (OR 2.6, CI 2.0-3.4). Significant predictors for pain recovery were male gender, younger age, cohabitation status, good self-rated health, good mental health, having no long standing disease, and having an annual income above 15,400 US dollars. PMID- 15109520 TI - Antiallodynic effects of intrathecally administered 5-HT(2C) receptor agonists in rats with nerve injury. AB - Intrathecal administration of serotonin type 2 (5-HT(2)) receptor agonists, alpha methyl-5-hydroxytryptamine maleate (alpha-m-5-HT) or (+/-)-1-(4-iodo-2,5 dimethoxyphenyl)-2-aminopropane hydrochloride (DOI), produces antiallodynic effects in a rat model of neuropathic pain. In the present study, we examined the antiallodynic effects of intrathecally administered agents which are selective for 5-HT(2C) receptors. Allodynia was produced by tight ligation of the left L5 and L6 spinal nerves, and was measured by applying von Frey filaments to the left hindpaw. Administration of the 5-HT(2C) receptor agonist, 6-chloro-2-(1 piperazinyl)-pyrazine (MK212; 3-100 microg), 1-(m-chlorophenyl)-piperazine (mCPP; 30-300 microg), or 1-(m-trifluoromethylphenyl)-piperazine (TFMPP; 30-300 microg), produced antiallodynic effects in a dose-dependent manner with no associated motor weakness. The ED(50) values of MK212, mCPP, and TFMPP were 39.2, 119.9, and 191.9 microg, respectively. Intrathecal pretreatment with the selective 5-HT(2C) receptor antagonist RS-102221 (30 microg) diminished the effects of the highest doses of 5-HT(2C) receptor agonists. The preferential 5-HT(2A) receptor antagonist ketanserin (30 microg) did not reverse the effects. In contrast to 5 HT(2C) receptor agonists, the antiallodynic effects of intrathecally administered alpha-m-5-HT (30 microg) and DOI (100 microg) were reversed by ketanserin, but not by RS-102221. These results indicate that 5-HT(2C) receptors have a role in spinal inhibition of neuropathic pain, and the effects produced by intrathecal administration of 5-HT(2C) receptor agonists are mediated by a mechanism different from that of alpha-m-5-HT or DOI, which seem to produce their effects through 5-HT(2A) receptors. PMID- 15109521 TI - Pain during mammography: the role of coping strategies. AB - This study examined the pain/discomfort ratings during mammography and their relationship to pain-coping strategies. Prior to their mammogram in order to assess how they cope with day-to-day pain experience 220 women completed an Iranian translation of the Coping Strategies Questionnaire (IR-CSQ). Immediately after mammography, all subjects completed a modified version of the IR-CSQ to assess their coping style with pain during mammography. Ratings of pain/discomfort during the mammogram were also collected, using a 100-mm Visual Analog Scale and a 6-point Pain/Discomfort Rating Scale (PRDS). Up to 92% of the women reported that the mammogram examination was painful. However, considerable variability in pain ratings was found, with some women reporting severe pain and others reporting little or no pain. While the ratings of coping effectiveness in facing day-to-day pain experience were not significantly related to mammography pain ratings, there was a significant association between the ratings of coping efficacy in facing mammography pain specifically and measures of mammography pain. Multiple hierarchical regression analyses revealed that while higher use of catastrophising and coping self-statements in facing mammography pain were predictors of more severe pain during mammography, higher use of ignoring pain sensations was predictive of less severe pain during mammography. The potential significance of these findings for identifying patients at risk of poor adjustment to chronic pain is discussed. PMID- 15109522 TI - Snake venom phospholipase A2s (Asp49 and Lys49) induce mechanical allodynia upon peri-sciatic administration: involvement of spinal cord glia, proinflammatory cytokines and nitric oxide. AB - Snakebites constitute a serious public health problem in Central and South America, where species of the lancehead pit vipers (genus Bothrops) cause the majority of accidents. Bothrops envenomations are very painful, and this effect is not neutralized by antivenom treatment. Two variants of secretory phospholipases A2 (sPLA2), corresponding to Asp49 and Lys49 PLA2s, have been isolated from Bothrops asper venom. These sPLA2s induce hyperalgesia in rats following subcutaneous injection. However, venom in natural Bothrops bites is frequently delivered intramuscularly, thereby potentially reaching peripheral nerve bundles. Thus, the present series of experiments tested whether these sPLA2s could exert pain-enhancing effects following administration around healthy sciatic nerve. Both were found to produce mechanical allodynia ipsilateral to the injection site; no thermal hyperalgesia was observed. As no prior study has examined potential spinal mechanisms underlying sPLA2 actions, a series of anatomical and pharmacological studies were performed. These demonstrated that both sPLA2s produce activation of dorsal horn astrocytes and microglia that is more prominent ipsilateral to the site of injection. As proinflammatory cytokines and nitric oxide have each been previously implicated in spinally mediated pain facilitation, the effect of pharmacological blockade of these substances was tested. The results demonstrate that mechanical allodynia induced by both sPLA2s is blocked by interleukin-1 receptor antagonist, anti-rat interleukin-6 neutralizing antibody, the anti-inflammatory cytokine interleukin-10, and a nitric oxide synthesis inhibitor (L-NAME). As a variety of immune cells also produce and release sPLA2s during inflammatory states, the data may have general implications for the understanding of inflammatory pain. PMID- 15109523 TI - Graded motor imagery is effective for long-standing complex regional pain syndrome: a randomised controlled trial. AB - Complex regional pain syndrome type 1 (CRPS1) involves cortical abnormalities similar to those observed in phantom pain and after stroke. In those groups, treatment is aimed at activation of cortical networks that subserve the affected limb, for example mirror therapy. However, mirror therapy is not effective for chronic CRPS1, possibly because movement of the limb evokes intolerable pain. It was hypothesised that preceding mirror therapy with activation of cortical networks without limb movement would reduce pain and swelling in patients with chronic CRPS1. Thirteen chronic CRPS1 patients were randomly allocated to a motor imagery program (MIP) or to ongoing management. The MIP consisted of two weeks each of a hand laterality recognition task, imagined hand movements and mirror therapy. After 12 weeks, the control group was crossed-over to MIP. There was a main effect of treatment group (F(1, 11) = 57, P < 0.01) and an effect size of approximately 25 points on the Neuropathic pain scale. The number needed to treat for a 50% reduction in NPS score was approximately 2. The effect of treatment was replicated in the crossed-over control subjects. The results uphold the hypothesis that a MIP initially not involving limb movement is effective for CRPS1 and support the involvement of cortical abnormalities in the development of this disorder. Although the mechanisms of effect of the MIP are not clear, possible explanations are sequential activation of cortical pre-motor and motor networks, or sustained and focussed attention on the affected limb, or both. PMID- 15109524 TI - Motor cortex stimulation for central and neuropathic pain (Letter regarding Topical Review by Brown and Barbaro). PMID- 15109526 TI - Evidence for heritability of pain in patients with traumatic neuropathy. PMID- 15109528 TI - Angiogenesis in acute and chronic lymphocytic leukemia. AB - The bone marrow microenvironment plays a crucial role in leukemogenesis. Recent studies suggest that its vascularity changes significantly during this process and that angiogenic factors are of major importance in leukemia. This review summarizes the literature concerning the relationship between angiogenesis and the progression of acute and chronic lymphocytic leukemia. It is becoming increasingly evident that agents which interfere with angiogenesis also block tumor progression and anti-angiogenic management has become a prominent aspect of pre-clinical and clinical assessment. Recent applications of anti-angiogenic agents which interfere with or block leukemia progression are reviewed. PMID- 15109529 TI - Thalidomide therapy for myelodysplastic syndromes: current status and future perspectives. AB - Thalidomide exerts in vitro heterogeneous biological effects on hematopoiesis which have supported its possible use in treating myelodysplastic syndromes (MDS). Some recent clinical trials have confirmed that thalidomide may improve anemia and, less frequently, other cytopenias, in a proportion of younger patients with low-risk MDS (11-56%, on intention-to-treat analysis). Of interest, erythroid responses may be achieved also in transfusion-dependent subjects with high serum levels of endogenous erythropoietin, a subset of MDS patients with little chance of responding to recombinant erythropoietin, alone or in combination with G-CSF. Older patients, however, often do not tolerate the drug even at very low doses. How thalidomide acts in MDS is not clear. Some data suggest several mechanisms possibly involving stimulation of erythropoiesis through activation of physiological compensative mechanisms and reduction of apoptosis. The combination of thalidomide with other molecules active on hematopoiesis and the use of more effective and less toxic analogs are currently under clinical investigation. PMID- 15109530 TI - Distinct cellular origins of primary effusion lymphoma with and without EBV infection. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is associated with three distinct lymphoproliferative disorders: primary effusion lymphoma (PEL), multicentric Castleman's disease (MCD) and germinotropic lymphoproliferative disorder (GLD). KSHV positive lymphocytes in GLD and in most cases of PEL are co-infected by Epstein-Barr virus (EBV) and these viral double positive cells harbour mutated rearranged immunoglobulin (Ig) genes, suggesting that they originate from germinal centre or post-germinal centre B-cells. In contrast, KSHV positive cells in MCD are invariably negative for EBV, do not carry Ig gene mutation and are believed to originate from naive IgMlambda expressing B-cells. Interestingly, one EBV negative PEL (BC3) also lacks Ig gene mutation, raising the question whether KSHV preferentially targets naive B-cells in the absence of EBV. We compared the cellular origin of PEL with and without EBV infection by analysis of Ig gene mutation. High molecular weight DNA from 17 PELs was subjected to PCR of the rearranged Ig heavy and light chain genes. Successful amplification was achieved from eight cases (four EBV positive and four EBV negative) and the PCR products were sequenced. All four EBV positive PEL showed variable levels of mutation in their rearranged V(H) or V(L) genes, ranging from 4 to 7%. In contrast, two of the four EBV negative PELs including BC3 displayed absence of mutation in their rearranged Ig genes. Our results indicate that EBV positive PELs are derived from germinal centre or post-germinal centre B-cells, whereas EBV negative PELs may originate from either germinal/post-germinal centre or naive B-cells. PMID- 15109531 TI - Allelotyping of gastrointestinal nasal-type NK/T-cell lymphoma. AB - Nasal-type natural killer/T-cell lymphoma (NKL) is a rare but distinct malignancy that often involves the mid-facial region and the gastrointestinal tract. This study is the first genome-wide allelotyping analysis on this rare lymphoma. We applied 382 microsatellite markers covering loci which spanned 22 autosomes to screen for allelic imbalances (AI) in six intestinal NKL. The most common chromosomal regions of allelic imbalances were found in 11p, 9q and 13q. Novel spots of allelic losses spots found at 2p21, 2q37.22, 18p11.21 and 18q12.1. In spite of presence of a few recurrent loci of imbalances, the allelotyping results show that NKL is heterogeneous. PMID- 15109532 TI - Comparison of M-FISH and conventional cytogenetic analysis in accelerated and acute phases of CML. AB - FISH and multicolor FISH (M-FISH) techniques have greatly enhanced the resolution of conventional cytogenetic analysis, thus enabling the identification of novel regions of rearrangement in hematological malignancies. We report on the analysis of cells from 24 chronic myelogenous leukemia (CML) patients, in either accelerated phase (14 cases) or blast crisis (10 cases) aimed at searching for previously unidentified additional abnormalities related to disease evolution. Indeed, in 6 of 24 cases (25%) M-FISH allowed a more precise description of chromosomal aberrations, the finding of cryptic rearrangements, characterization of markers, identification of additional material and a better interpretation of complex aberrations. However, new recurrent aberration did not emerge from M-FISH analysis. PMID- 15109533 TI - Phase II trial of cladribine and cytarabine in relapsed or refractory myeloid malignancies. AB - To evaluate the efficacy of cladribine and cytarabine in children with relapsed or refractory myeloid malignancies, we administered cytarabine (200 mg/m2 per day) by continuous subcutaneous infusion and cladribine (8.9 mg/m2 per day) by continuous intravenous infusion concomitantly for 5 days to nine patients younger than 21 years. After one course, five patients had no response, two patients had partial responses, one had stable disease, and one had progressive disease. Two patients received a second course: one patient had stable disease after one course and progressive disease after the second; another patient had a partial response after one course and no response after the second. Despite the efficacy of the cladribine and cytarabine regimen in treating newly diagnosed acute myeloid leukemia (AML) in a previously reported study, the combination was not effective for relapsed or refractory childhood AML. PMID- 15109534 TI - A fludarabine, topotecan, and cytarabine regimen is active in patients with refractory acute myelogenous leukemia. AB - As fludarabine, topotecan and cytarabine (ara-C) are effective in acute myeloid leukemia (AML), a pilot study of these three agents combined (FTA) was conducted. FTA consisted of topotecan 1.25 mg/m2 by CIV days 1-5, fludarabine 15 mg/m2 and ara-C 0.5 g/m2 IV, BID, on days 2-6. Seventeen patients (6 primary resistant, 11 relapsed) with AML received 33 courses of FTA. Six patients (35%) achieved complete remission. Seven patients (41%) developed grade 3 or 4 diarrhea. FTA was effective and warrants further study in patients with refractory AML. PMID- 15109535 TI - Prognostic significance of Fas (CD95) and TRAIL receptors (DR4/DR5) expression in acute myelogenous leukemia. AB - We analyzed the clinical significance of the expression of the Fas (CD95) and TNF related apoptosis-inducing ligand (TRAIL) receptors, the death receptors (DR) 4 and 5, by leukemic blasts in 29 patients with acute myelogenous leukemia (AML). CD95 was positive in 18 patients (62%). The DR4 and DR5 receptors were positive in 20 patients (69%) and 29 (100%), respectively. CD95 positivity was not correlated with cytogenetic abnormalities. Complete remission (CR) rate was not significantly different according to the expression of the CD95 or TRAIL receptors. Relapse-free survival was significantly prolonged in patients with CD95-positive AML cells compared with patients with CD95-negative AML cells (73% versus 38% at 3 years; P = 0.047). TRAIL receptors did not show correlation with other clinical parameters. PMID- 15109536 TI - Evaluation of BCRP and MDR-1 co-expression by quantitative molecular assessment in AML patients. AB - Expression of two MDR genes, BCRP and MDR1, was evaluated by real-time PCR technique in 51 AML patients. Fifty-six percent expressed the BCRP gene, with the 48.2% showing intermediate levels. Eighty-eight percent expressed the MDR1, with 23.8% of cases at high expression. A significant correlation between BCRP and MDR1 values was found by regression analysis. Either levels of BCRP or MDR1 did not correlate with clinical characteristics of patients at diagnosis. PMID- 15109537 TI - C-kit receptor expression in acute leukemias-association with patient and disease characteristics and with outcome. AB - We hypothesize that c-kit expression may be associated with disease-specific features and have prognostic value in acute leukemias. In acute lymphocytic leukemia (ALL), higher levels of c-kit expression predicted lower complete response (CR) rates, suggesting that these patients may benefit from acute myelogenous leukemia (AML) therapy. Despite a negative association with the Philadelphia-chromosome, there was no correlation with disease-free survival (DFS) in CR. In AML, c-kit was associated with older age and cytogenetic abnormality t(-5, -7). Consequently higher levels of c-kit predicted lower CR rates. However, after accounting for these covariates, multivariate analysis indicates that higher c-kit expression predicts higher CR rates, although there was no effect on DFS in CR. PMID- 15109538 TI - Methylation profiling in multiple myeloma. AB - BACKGROUND: We analysed the methylation status of a panel of 10 genes including p15, p16, DAPK, p73, VHL, E-CAD, MGMT, RARbeta, RIZ1, and ER. METHODS: The gene promoter methylation status was studied by methylation-specific polymerase chain reaction (MSP) with primers for methylated (M-MSP) and unmethylated (U-MSP) DNA in the bone marrow of 13 patients with myeloma, and one patient with plasmacytoma. RESULT: None of the 10 genes tested were methylated in eight normal bone marrow samples. For the positive control, the sensitivity of M-MSP ranged from 1 x 10(-2) for E-CAD and MGMT, to 1 x 10(-4) for p73. Of the eight diagnostic myeloma marrow samples, hypermethylation of p15, p16, E-CAD, DAPK and ER occurred in six (75%), four (50%), seven (87.5%), eight (100%), and six (75%) patients. Similarly, of the five samples from patients who progressed from plateau phase, hypermethylation of p15, p16, E-CAD, DAPK, and ER occurred in five (80%), two (40%), five (100%), five (100%), and three (60%). None of the cases had hypermethylation of RIZ1, p73, VHL, RARbeta, and MGMT. At diagnosis, all patients had concurrent hypermethylation of at least three genes, and five (62%) had concurrent methylation of four or more genes. One patient with plasmacytoma had methylation of E-CAD, ER, and DAPK. CONCLUSION: p15, p16, ER, DAPK, and E-CAD (but not RARbeta, p73, VHL, RIZ1, and MGMT) were frequently methylated in MM at both diagnosis and disease progression. Future studies of larger scale are needed to identify the genes responsible for disease progression. PMID- 15109539 TI - Peroxisome proliferator-activated receptor gamma ligands induce growth inhibition and apoptosis of human B lymphocytic leukemia. AB - This study examined the expression and structural intactness of peroxisome proliferator-activated receptor gamma (PPARgamma) in human acute lymphocytic leukemia (ALL) cells and determined the effect of PPARgamma ligands on growth and apoptosis of these cells. We noted that all lymphocytic leukemia cell lines expressed PPARgamma and no PPARgamma mutations were found in these cell lines as indicated by SSCP analysis. Effect of the PPARgamma ligands on the proliferation, differentiation and apoptosis of B type ALL cells was further examined. Treatment of these cells with the PPARgamma ligands Pioglitazone (PGZ) and 15-deoxy-delta (12,14)-prostaglandin J2 (15d-PGJ2) resulted in growth inhibition in a dose dependent manner which was associated with a G1 to S cell cycle arrest. However, this effect appeared to be PPARgamma-independent since several PPARgamma antagonists could not reverse this effect. No differentiation was induced by this treatment. Four out of five cell lines underwent apoptosis after culture with the PPARgamma ligands. This effect was partially caspase-dependent because a pan caspase inhibitor partially reversed this effect. In conclusion, our results suggest that PPARgamma ligands may offer a new therapeutic approach to aid in the treatment of ALL. PMID- 15109540 TI - Combination of all-trans retinoic acid and lithium chloride surmounts a retinoid differentiation block induced by expression of Scl and Rbtn2 transcription factors in myeloid leukemia cells. AB - We have previously shown that forced expression of the transcription factor Scl in WEHI-3B D(+) cells prevents ATRA-induced cell differentiation. We now find that the overexpression of Rbtn2 also interferes with induction of differentiation by ATRA. Addition of LiCl to ATRA treatment restored the capacity of both Scl- and Rbtn2-expressing cells to respond to the retinoid in a synergistic manner. Similar results were obtained with Scl-transfected HL60 cells where its expression diminished responsiveness to ATRA. These findings suggest that if Scl and/or Rbtn2 are involved in the non-responsiveness of AML patients to ATRA-induced differentiation, addition of LiCl may reverse insensitivity. PMID- 15109541 TI - Imatinib mesylate (STI571) prevents the mutator phenotype of Bcr-Abl in hematopoietic cell lines. AB - Progression of CML from chronic phase to blast crisis is accompanied by accumulating genetic alterations. To analyze whether this abnormality can be prevented by inhibition of Bcr-Abl, we measured the frequency of spontaneous and irradiation-induced HPRT mutations in cells treated with or without imatinib mesylate (Gleevec, STI571). Imatinib treatment of cells expressing Bcr-Abl reversed the mutation frequency to a value comparable to that of Bcr-Abl negative cells. Experiments with a Bcr-Abl deletion mutant indicate that in addition to the kinase activity, protein-protein interactions are required for induction of the mutator phenotype by Bcr-Abl. PMID- 15109542 TI - Domains involved in ETO and human N-CoR interaction and ETO transcription repression. AB - The (8;21) translocation between the AML1 and ETO genes is seen in approximately 12-15% of all acute myeloid leukemia (AML) and is a frequently observed nonrandom genetic alteration associated with AML. The ETO moiety was shown to interact with the nuclear receptor co-repressor (N-CoR) complex, which includes mSin3A and the histone deacetylase, HDAC1. Repression of AML1-responsive hematopoietic genes by AML1-ETO and the N-CoR complex may play a mechanistic role in t(8;21) leukemogenesis. In order to characterize the interaction between ETO and N-CoR, mutants of either protein were constructed and tested for binding in both yeast two-hybrid and immunoprecipitation assays. We found that two domains of human N CoR, amino acid residues 988-1126 and 1551-1803, were necessary for interaction with ETO. Previously, we and other investigators had reported that two unusual zinc finger motifs at the C-terminus of ETO mediated binding to N-CoR. Here, using mammalian two-hybrid assays, we found that transcription repression by ETO was substantially decreased when either zinc finger motif of ETO is deleted or mutated. In addition, we identified a second transcription repression domain located between residues 275 and 487. Characterization of the ETO interaction domains within human N-CoR and of the transcription domains of ETO is a first step in designing targeted molecular therapy for t(8;21) AML. PMID- 15109543 TI - Differences and similarities in kinetics of BCR-ABL transcript levels in CML patients treated with imatinib mesylate for chronic or accelerated disease phase. AB - Kinetics of BCR-ABL transcript levels were determined in 19 patients with chronic myeloid leukemia (CML) treated with imatinib for chronic (CP) or accelerated phase (AP). Patients could be divided into three groups with: (1) a sharp and sustained decrease in BCR-ABL transcript level reaching 0.1-0.002% (only CP); (2) an early BCR-ABL overexpression up to 2500% (only AP); and (3) a stable trend with BCR-ABL values between 10 and 100% (CP, AP). In group 1, relapses were not developed within the follow-up; in group 2, patients progressed to blast crisis; in group 3, BCR-ABL overexpression appeared after 12 months in some patients and disease relapses were found 2-16 weeks later. It is summarized that BCR-ABL transcript kinetics clearly characterize responses to imatinib treatment and are highly predictive for disease progression. PMID- 15109544 TI - Inefficacy of imatinib-mesylate in sporadic, aggressive systemic mastocytosis. PMID- 15109545 TI - Microsatellite mutations of transforming growth factor-beta receptor type II and caspase-5. PMID- 15109547 TI - Audiometric changes associated with the treatment of uncomplicated falciparum malaria with co-artemether. AB - Animal studies have demonstrated artemisinin brain stem toxicity with auditory centres being especially affected; there has, to date, been no evidence of such toxicity with oral artemisinins in humans. Subjects working at a construction site in Mozambique had audiometric assessments taken on joining and leaving the project. Subjects with uncomplicated malarias received co-artemether (artemether lumefantrine) (n = 150) while age-, gender-, weight- and race-matched controls (n = 150) neither suffered malaria nor received antimalarial therapy. Hearing thresholds were measured at predefined frequencies in treated subjects and controls. Subjects receiving co-artemether had a significantly greater heating loss than controls at all frequencies except 250 Hz and 500 Hz (P values ranging from <0.001 to 0.04, Mann-Whitney U). Mean changes at the different frequencies in subjects ranged from -6.50 dB (95% CI -8.19 to -4.81) [at 1kHz frequency] to 0.07 dB (95% CI -2.19 to 2.05) [at 6 kHz frequency]. Mean changes in the control group ranged from -4.20 dB (95% CI -5.97 to -2.43) [at 1 kHz frequency] to +2.7 dB (95% CI -0.93 to 4.47) [at 6 kHz frequency]. Treatment of uncomplicated malaria with co-artemether is associated with hearing loss, possibly from synergy between potentially ototoxic agents in combination. The safety and neurotoxicity of artemesinins and other endoperoxides needs to be more fully evaluated. PMID- 15109549 TI - Plague: the dreadful visitation occupying the human mind for centuries. AB - Plague is one of mankind's greatest scourges, which has swept away millions of people over the centuries. The first available record of the occurrence of this calamity, in humans, is from the Bible, in 1000 bc, in the city of Ashdod. The first definitely identified pandemic originated in Egypt in ad 542 (the Justinian Plague) and is estimated to have caused 100 million deaths. The second one, lasting for three centuries and claiming over 25 million lives appeared in 1334 in China spreading to many spots on the globe. The third pandemic occurred in Europe from the fifteenth to eighteenth century. The current pandemic began around 1860, in the Chinese province Yunnan; it reached Hong Kong in 1894 killing 100 000 individuals. Within 20 years the disease spread from southern Chinese ports throughout the world resulting in more than 10 million deaths. Since the discovery of the causative agent in 1894, there have been remarkable advancements in immunoprophylaxis and chemoprophylaxis. However, the disease is still active in Africa, in Asia and in Americas and has been classified as a currently re emerging disease. A 'Plague-free World' will probably remain a dream for an indefinite period. PMID- 15109550 TI - Protocol and monitoring to improve snake bite outcomes in rural Ghana. AB - A study was conducted in Mathias Hospital, Yeji, an area of Ghana, where snake bite cases are an important cause of morbidity and mortality, with a case fatality rate of 11% (8/72). Case management difficulties included uncertainty about the assessment of the severity of envenoming, the dosage of antivenom, and the response to treatment. An intervention with several components was introduced: development of a treatment protocol, staff training, monitoring of compliance and patient education. During a 33-month post-intervention period there was excellent protocol compliance, fewer snake bite complications, and a fall in mortality rate to 1.3% (3/238) compared with a 15-month baseline review. There was a 50% increase in snake bite admissions and fewer delays. To improve snake bite outcomes in comparable settings, particularly if inexperienced staff are involved in care, we recommend a similar quality assurance project, involving case review and use of a treatment protocol with monitoring of compliance to sustain an improved approach. PMID- 15109551 TI - Cross-sectional study of specific antibodies to a polymorphic Plasmodium falciparum antigen and of parasite antigen genotypes in school children on the slope of Mount Cameroon. AB - To investigate relationships between Plasmodium falciparum parasitaemia, parasite genotypes, and specific anti-parasite antibodies, 244 school children (aged 4 to 16 years) were studied in April/May 2002, the peak malaria transmission season in Buea, Cameroon. Antibody reactivities were analysed by ELISA using an array of recombinant antigens representing different sequences from the polymorphic block 2 region of the merozoite surface protein 1 (MSP1), and the blood samples that were slide-positive for P. falciparum were genotyped for msp1 block 2 alleles. The prevalence of antibodies to the specific MSP1 block 2 antigens was significantly higher in children at one particular school (situated at the lowest altitude) compared to the others, although the prevalence of infection or particular parasite genotypes did not differ. Thus, at a population level, the prevalence of these antibodies does not simply reflect prevalence of parasites, but rather may be due to differences in the incidence of past infections. However, there were weak positive associations between specific antibody reactivity and the presence of the corresponding allele in the blood of individuals (statistically significant for the MAD20-type allele of block 2), indicating that antibody specificities are to some extent determined by current parasite infections. PMID- 15109552 TI - Fatal type A botulism in South Africa, 2002. AB - Although wildfowl and domestic livestock botulism has been recognized as a problem in southern Africa, very few human cases have ever been described in the region. In late February 2002, two siblings aged eight and 12 years developed acute flaccid paralysis and died. Mouse bioassays revealed the presence of type A botulinum toxin in the serum of both children, and in the retrieved remains of the implicated food. The implicated vehicle of the toxin was tinned fish in tomato sauce, commercially produced in South Africa. Type A Clostridium botulinum was cultured from the food. The most likely scenario was that corrosion damage had allowed entry of environmental organisms, including Clostridium botulinum, to the tinned food. This is the first outbreak of human type A botulism in southern Africa to be documented, and the first fatal outbreak described; previous human cases in this region have involved type B botulinum toxin, which tends to produce milder disease. A few other outbreaks elsewhere in Africa have been published, the most extensive being a type E epidemic in Egypt. Commercially tinned products were not involved in any of those outbreaks. PMID- 15109553 TI - Emergence of Schistosoma mansoni in the Niger River valley, Niger. AB - Urinary and intestinal schistosomiasis are both endemic in Niger with a dominance of Schistosoma haematobium. This study pointed out the appearance of the infection to S. mansoni in the Niger River valley, where it was until then unknown in Niger. In one year the prevalence increased from 5.9 to 19.5%. PMID- 15109554 TI - Leishmaniasis due to Leishmania tropica MON-102 in a new Moroccan focus. AB - In Morocco, between March and April 2002, large numbers of human leihsmaniasis cases were detected during a survey at Zouagha My Yacoub province in Fes State. Among 95 cases, 76 were positive by direct observation of Giemsa-stained smears. Sixteen stocks were isolated in NNN medium and identified as Leishmania tropica MON-102, using isoenzyme techniques on starch gel electrophoresis. PMID- 15109555 TI - Limited influence of haemoglobin variants on Plasmodium falciparum msp1 and msp2 alleles in symptomatic malaria. AB - Haemoglobin (Hb) S, HbC, and alpha(+)-thalassaemia confer protection from malaria. Accordingly, these traits may influence the multiplicity of infection (MOI) of Plasmodium falciparum and the presence of distinct parasite genotypes. In 840 febrile children in northern Ghana, we typed the P. falciparum merozoite surface protein genes (msp1, msp2) and examined effects of the Hb variants on MOI and parasite diversity. HbAC, HbAS, heterozygous, and homozygous alpha(+) thalassaemia occurred in 21, 5, 29 and 4% of the children, respectively. Plasmodium falciparum was detected in 95%. The haemoglobinopathies did not influence MOI, nor did the Hb type bias the distribution of the msp allelic families. However, IC type parasites were most common among patients with homozygous alpha(+)-thalassaemia (93%), less frequent in heterozygotes (89%), and least frequent in alpha-globin normal children (84%, P(chi2 trend) = 0.03). The opposite was seen for Mad20 type parasites (34%, 47%, 53%, P(chi2 trend) = 0.02). Only a few of the 72 individual msp alleles were selected by the haemoglobinopathies. HbC and alpha(+)-thalassaemia are frequent in northern Ghana. In symptomatic children, the effect of Hb variants on parasite multiplicity and diversity appears to be limited. This may reflect an actual lack of influence or indicate abrogation in symptomatic malaria. PMID- 15109556 TI - Decreasing clinical efficacy of chloroquine in Ankazobe, Central Highlands of Madagascar. AB - The clinical efficacy of chloroquine was assessed in the Primary Health Centre of Ankazobe, Central Highlands of Madagascar. This study shows an increase in the level of chloroquine resistance with the appearance of early treatment failures and RIII resistance. Furthermore, the prevalence of clinical treatment failures is approaching the level of 25%, at which WHO recommends a change of first-line drug. PMID- 15109557 TI - A simple device to maintain in vitro cultures of Leishmania in tropical countries. AB - A thermo-stable clay device to maintain in vitro Leishmania cultures in tropical countries within the optimal temperature range of 22 to 27 degrees C was developed. This being simple and economical could immensely benefit diagnosis and research on leishmaniasis in these countries. PMID- 15109558 TI - Randomized controlled trial of a traditional preparation of Artemisia annua L. (Annual Wormwood) in the treatment of malaria. AB - The Chinese medicinal plant Artemisia annua L. (Annual Wormwood) contains the antimalarial compound artemisinin. The locally grown herb may offer an additional tool for the control of malaria, especially in poor countries where modern antimalarial drugs are often unavailable. In an open, randomized, controlled pilot trial, we investigated the efficacy and safety of traditional tea preparations of Artemisia annua in the treatment of uncomplicated malaria. Treatment resulted in a quick resolution of parasitaemia and of clinical symptoms. After 7 d of medication, cure rates were on average 74% for the Artemisia preparations compared with 91% for quinine. However, recrudescence rates were high in the Artemisia groups. Therefore, monotherapy with Artemisia annua L. cannot be recommended as alternative to modern antimalarials, but may deserve further investigation. PMID- 15109559 TI - Seasonal variation in hyperparasitaemia and gametocyte carriage in patients with Plasmodium falciparum malaria on the Thai-Burmese border. AB - Between January 2000 and December 2002 monthly rainfall was correlated with the proportion of patients with hyperparasitaemic Plasmodium falciparum malaria and with the proportion of patients with P. falciparum gametocytes. During the observation period 6953 cases of P. falciparum malaria were treated at the Shoklo Malaria Research Unit in Maela refugee camp on the Thai-Burmese border. Three hundred and seventy-five of these patients had >/=4% of parasitized red blood cells. Although there were more monthly malaria cases in the rainy season, rainfall was negatively correlated with the proportion of patients with hyperparasitaemia (Spearman's rho = -0.59, P < 0.001 ), and the proportion of gametocyte carriers among P. falciparum cases, (Spearman's rho = -0.39, P = 0.018). After controlling for age and the origin of the patient, the odds ratio for developing hyperparasitaemia during the dry season was 1.6 (95% CI 1.14-2.2; P = 0.006). The adjusted odds ratio for gametocyte carriage during the dry season was 1.3 (95% CI 1.03-1.6; P = 0.02). Migrations, changes in transmission patterns, the haematological burden of cumulative infections, and ultraviolet immunosuppression are discussed as potential explanations for these observations. PMID- 15109560 TI - Epidermal keratinocytes: regulation of multiple cell phenotypes by multiple protein kinase C isoforms. AB - Squamous cells form the outermost layers of the epidermis, and though they are readily discarded from the tissue, they serve a vital water barrier function while in the stratum corneum. The generation of cornified or squamous keratinocytes involves a complex, multi-step differentiation process that insures the proper physical and immunological barrier functions of the epidermis are maintained. The regulation of keratinocyte terminal differentiation is influenced by a large number of signaling pathways. This article will review some recent findings regarding the roles of the protein kinase C (PKC) family in normal keratinocyte differentiation, as well as their involvement in skin diseases, especially skin cancer. PMID- 15109561 TI - Lymphatic smooth muscle: the motor unit of lymph drainage. AB - Embedded into the wall of collecting lymphatic vessels and trunks, the lymphatic smooth muscles are cardinal to the functions of the lymphatic system. Their intrinsic contractile property--the intrinsic lymph pump--through rhythmical and phasic contractions of the vessels, represents the principal mechanism by which lymph flow is generated. Through changes in tonic constrictions, lymphatic smooth muscles also modulate lymph flow resistance. Lymphatic smooth muscles are sensitive to physical and chemical stimuli, mediating changes in their activity and modulating lymphatic drainage. Because lymphatic smooth muscles play such an important role in fluid transport, their dysfunction may be a component of many inflammatory disease states. This review presents recent findings on the physiology and cellular biology of lymphatic smooth muscles and discusses the importance of these cells for the function of the lymphatic system in physiological and pathophysiological situations. PMID- 15109562 TI - Small cell lung cancer: the importance of the extracellular matrix. AB - Lung cancer is the leading cause of cancer deaths in the UK and the small cell lung cancer (SCLC) phenotype is the most aggressive form of this disease, with a high metastatic potential and the development of resistance to chemotherapy. Evidence now suggests that these features may be due to important links between the cancer cells and proteins in their local extracellular matrix (ECM). This article reviews the evidence for a chemoprotective effect of extracellular matrix in small cell lung cancer and discusses the importance of integrin-mediated signalling pathways in this setting. PMID- 15109563 TI - Mammalian transforming growth factor-betas: Smad signaling and physio pathological roles. AB - Since its discovery in the early 1980s, transforming growth factor-beta (TGF beta) has emerged as a family of growth factors involved in essential physiological processes, including embryonic development, differentiation, tissue repair and cell growth control. Knockout experiments for the three mammalian isoforms of TGF-betas in mice have demonstrated their importance in regulating inflammation and tissue repair. Also, TGF-beta has been implicated in the pathogenesis of human diseases, including tissue fibrosis and carcinogenesis where, in the latter case, it may exert both tumor suppressor and pro-oncogenic activities depending on the stage of the tumor. Cellular signaling by TGF-beta family members is initiated by the assembly of specific cell surface serine/threonine kinase type receptors that activate transcription factors of the Smad family. PMID- 15109564 TI - NFAT and NF-kappaB factors-the distant relatives. AB - NFAT and NF-kappaB proteins are members of a superfamily of transcription factors whose activity plays a crucial role in the activation, proliferation and apoptosis of lymphocytes. Both types of factors share a number of properties, including similar DNA binding domains and rapid nuclear translocation upon antigenic stimulation. While NF-kappaBs control both innate and adaptive immune responses, NFATs control the adaptive immune system which emerged-in parallel with the appearance of the NFAT family-in jawed fish. However, NFATs and NF kappaBs differ remarkably in their function. Whereas NFATs support activation induced cell death (AICD) of T and B cells, NF-kappaB proteins frequently exert a strong anti-apoptotic effect on lymphocytes and other cells. While the anti apoptotic activity of NF-kappaBs contributes to their oncogenic capacity, the pro apoptotic activity favors NFATs as tumor suppressors in lymphoid cells. PMID- 15109566 TI - Prostanoids and prostanoid receptors in signal transduction. AB - Prostanoids are arachidonic acid metabolites and are generally accepted to play pivotal functions in amongst others inflammation, platelet aggregation, and vasoconstriction/relaxation. Inhibition of their production with, for instance, aspirin has been used for over a century to combat a large variety of pathophysiological processes, with great clinical success. Hence, the cellular changes induced by prostanoids have been subject to an intensive research effort and especially prostanoid-dependent signal transduction has been extensively studied. In this review, we discuss the impact of the five basic prostanoids, TxA(2), PGF(2alpha), PGE(2), PGI(2), and PGD(2), via their receptors on cellular physiology. These inflammatory lipids may stimulate serpentine plasma membrane localized receptors, which in turn affect major signaling pathways, such as the MAP kinase pathway and the protein kinase A pathway, finally resulting in altered cellular physiology. In addition, prostanoids may activate the PPARgamma members of the steroid/thyroid family of nuclear hormone receptors, which act as transcription factors and may thus directly influence gene transcription. Finally, evidence exists that prostanoids act as second messengers downstream of mitogen receptor activation, mediating events, such as cytoskeletal changes, maybe via direct interaction with GTPase activating proteins. The final cellular reaction to prostaglandin stimulation will most likely depend on combined effects of the above-mentioned levels of interaction between prostaglandins and their cellular receptors. PMID- 15109565 TI - The effects of connexin phosphorylation on gap junctional communication. AB - Gap junctions are specialized membrane domains composed of collections of channels that directly connect neighboring cells providing for the cell-to-cell diffusion of small molecules, including ions, amino acids, nucleotides, and second messengers. Vertebrate gap junctions are composed of proteins encoded by the "connexin" gene family. In most cases examined, connexins are modified post translationally by phosphorylation. Phosphorylation has been implicated in the regulation of gap junctional communication at several stages of the connexin "lifecycle", such as the trafficking, assembly/disassembly, degradation, as well as, the gating of gap junction channels. Since connexin43 (Cx43) is widely expressed in tissues and cell lines, we understand the most about how it is regulated, and thus, connexin43 phosphorylation is a major focus of this review. Recent reports utilizing new methodologies combined with the latest genome information have shown that activation of several kinases including protein kinase A, protein kinase C, p34(cdc2)/cyclin B kinase, casein kinase 1, mitogen activated protein (MAP) kinase and pp60(src) kinase can lead to phosphorylation at 12 of the 21 serine and two of the six tyrosine residues in the C-terminal region of connexin43. In several cases, use of site-directed mutants of these sites have shown that these specific phosphorylation events can be linked to changes in gap junctional communication. PMID- 15109567 TI - Regulation of tight junctions and loss of barrier function in pathophysiology. AB - The mechanism by which epithelial and endothelial cells interact to form polarized tissue is of fundamental importance to multicellular organisms. Dysregulation of these barriers occurs in a variety of diseases, destroying the normal cellular environments and leading to organ failure. Increased levels of growth factors are a common characteristic of diseases exhibiting tissue permeability, suggesting that growth factors play a direct role in elevating permeability. Of particular concern for this laboratory, increased expression of vascular endothelial growth factor may enhance vascular permeability in diabetic retinopathy, leading to vision impairment and blindness. However, the mechanism by which growth factors increase permeability is unclear. Polarized cells form strong barriers through the development of tight junctions, which are specialized regions of the junctional complex. Tight junctions are composed of three types of transmembrane proteins, a number of peripheral membrane structural proteins, and are associated with a variety of regulatory proteins. Recent data suggest that growth factor-stimulated alterations in tight junctions contribute to permeability in a variety of disease states. The goal of this review was to elucidate potential mechanisms by which elevated growth factors elicit deregulated paracellular permeability via altered regulation of tight junctions, with particular emphasis on the tight junction proteins occludin and ZO-1, protein kinase C signaling, and endocytosis of junctional proteins. Understanding the molecular mechanisms underlying growth factor-mediated regulation of tight junctions will facilitate the development of novel treatments for diseases such as brain tumors, diabetic retinopathy and other diseases with compromised tight junction barriers. PMID- 15109568 TI - Polyamine depletion switches the form of 2-deoxy-D-ribose-induced cell death from apoptosis to necrosis in HL-60 cells. AB - Our previous studies demonstrated that intracellular polyamine depletion blocked HL-60 cell apoptosis triggered by exposure to 2-deoxy-d-ribose (dRib). Here, we have characterized the intracellular events underlying the apoptotic effects of dRib and the involvement of polyamines in these effects. Treatment of HL-60 cells with dRib induces loss of mitochondrial transmembrane potential, radical oxygen species production, intracellular glutathione depletion and translocation of Bax from cytosol to membranes. These effects are followed by cell death. However, the mode of cell death caused by dRib depends on intracellular levels of polyamines. d-Rib-treated cells with normal polyamine levels, progressing through the G(1) into the S and G(2)/M phases, undergo apoptosis, while in polyamine-depleted cells, being blocked at the G(1) phase, cell death mechanisms are switched to necrosis. The present study points to a relationship between the cell cycle distribution and the mode of cell death, and suggests that the level of intracellular spermidine, essential to cell cycle progression, may determine whether a cell dies by apoptosis or necrosis in response to a death stimulus. PMID- 15109569 TI - Serum concentration modifies amplitude and kinetics of voltage-gated Na+ current in the Mat-LyLu cell line of rat prostate cancer. AB - Voltage-gated Na+ channel (VGSC) expression has previously been shown to be upregulated in strongly metastatic prostate cancer cells (rat and human) and its activity shown to potentiate a variety of cellular behaviours integral to the metastatic cascade. However, the mechanism(s) responsible for the Na+ channel upregulation is not known. As a step towards evaluating the role of the extracellular biochemical environment in this regard, we have determined the effects of serum concentration on characteristics of Na+ channel expressed in the strongly metastatic Mat-LyLu rat prostate cancer cell line. Whole-cell patch clamp recording techniques were used to study the effects of serum concentrations, above and below the normal 1%. Both the amplitude and the kinetics of the currents were analysed. The following results were obtained: (1) Adding 1% foetal calf serum to cells starved of serum for 24h increased Na+ current density; however, increasing serum concentration further (to 5%) caused a reduction. (2) Serum-free medium produced Na+ currents with slower kinetics of activation (time to peak) and inactivation (exponential decay). (3) Increased serum concentration (a) shifted steady-state inactivation to more positive potentials without affecting conductance and (b) increased tetrodotoxin sensitivity. It is concluded that serum concentration is an important determinant of the Na+ channel characteristics leading to possible transcriptional and post translational modifications of channel expression and/or activity. Experiments are now needed to determine which constituents (protein hormones, growth factors, etc.) are responsible for these effects. PMID- 15109570 TI - No existence of translocus balancer to coordinate the expression and regulation of human hemoglobin genes in transgenic mice study. AB - All mammals use hemoglobin (Hb) to transport oxygen. Each Hb molecule is a tetramer of two pairs of unlike globin polypeptide chains. Equal amount of subunit globin chains derived from the corresponding alpha- and beta-like genes can always result during development though the two separate gene clusters are located on two different chromosomes and spatially transcribed within different nuclear domains. Disturbance of this balance will result in degradation or precipitation of the excessive globin chains, which is the character of various thalassemic syndromes. In previous studies, we had established two kinds of bacterial artificial chromosome (BAC) mediated transgenic mouse models, which contain respectively the entire human alpha- and beta-globin cluster. Here, we investigated the regulatory relationship between the two clusters by interbreeding these two kinds of transgenic mice. The levels of human alpha- and beta-mRNA in the various hybrid lines reflect the levels in the original transgenic lines that contain either the alpha- or beta-globin cluster alone. The results suggested that there is no apparent cross talk or regulatory interaction between the two human globin clusters in transgenic mice. PMID- 15109571 TI - Spectrophotometric analysis of the protective effect of ascorbate against spontaneous oxidation of tetrahydrobiopterin in aqueous solution: kinetic characteristics and potentiation by catalase of ascorbate action. AB - Tetrahydrobiopterin (BH(4)) is oxidized by O(2) readily in aqueous solutions and physiological concentrations of ascorbate have been shown to inhibit this reaction. In order to gain insight into the mechanism of ascorbate effect, a spectrophotometric analysis was applied for the study of the time course of BH(4) oxidation in the presence of various concentrations of ascorbate and the effect of various temperatures on the apparent second-order rate constant of BH(4) oxidation (k(ox)) in the presence or absence of catalase. In 100 micromol/l concentration, ascorbate alone prolonged the half-life time of 36 micromol/l BH(4) 1.4-fold whereas in the presence of catalase 1.85-fold. In the presence of catalase ascorbate decreased the value of k(ox) to 51 +/- 0.67%, whereas in the absence of it only to 64 +/- 0.77% of control (P < 0.01). The extent of ascorbate effect was not dependent on temperature, at least between 22 and 37 degrees C, either in the presence or absence of catalase. In the absence of catalase the apparent Arrhenius activation energies: 57.02 +/- 0.09 kJ/mol (-ascorbate) and 56.77 +/- 2.21 kJ/mol (+ascorbate) whereas in the presence of catalase: 62.72 +/- 1.37 kJ/mol (-ascorbate) and 59.93 +/- 2.84 kJ/mol (+ascorbate, mean +/- S.E.M., n=3) were obtained. The study shows that catalase potentiates the BH(4) stabilizing effect of ascorbate. It is concluded that removal of H(2)O(2) generated from BH(4) during oxidation by O(2) prevents a decrease of ascorbate concentration, and in the presence of ascorbate the pacemaker step in the overall reaction is the oxidation of BH(4) and not the reduction of the quinonoid BH(2) back to BH(4) by ascorbate. PMID- 15109572 TI - The purification and characterisation of novel dipeptidyl peptidase IV-like activity from bovine serum. AB - The discovery of a potentially novel proline-specific peptidase from bovine serum is presented which is capable of cleaving the dipeptidyl peptidase IV (DPIV) substrate Gly-Pro-MCA. The enzyme was isolated and purified with the use of Phenyl Sepharose Hydrophobic Interaction, Sephacryl S-300 Gel Filtration, and Q Sephacryl Anion Exchange, producing an overall purification factor of 257. SDS PAGE resulted in a monomeric molecular mass of 158kDa while size exclusion chromatography generated a native molecular mass of 328kDa. The enzyme remained active over a broad pH range with a distinct preference for a neutral pH range of 7-8.5. Chromatofocusing and isoelectric focusing (IEF) revealed the enzyme's isoelectric point to be 4.74. DPIV-like activity was not inhibited by serine protease inhibitors but was by the metallo-protease inhibitors, the phenanthrolines. The enzyme was also partially inhibited by bestatin. Substrate specificity studies proved that the enzyme is capable of sequential cleavage of bovine beta-Casomorphin and Substance P. The peptidase cleaved the standard DPIV substrate, Gly-Pro-MCA with a K(M) of 38.4 microM, while Lys-Pro-MCA was hydrolysed with a K(M) of 103 microM. The DPIV-like activity was specifically inhibited by both Diprotin A and B, non-competitively, generating a K(i) of 1.4 x 10(-4) M for both inhibitors. Ile-Thiazolidide and Ile-Pyrrolidide both inhibited competitively with an inhibition constant of 3.7 x 10(-7) and 7.5 x 10(-7) M, respectively. It is concluded that bovine serum DPIV-like activity share many biochemical properties with DPIV and DPIV-like enzymes but not exclusively, suggesting that the purified peptidase may play an important novel role in bioactive oligopeptide degradation. PMID- 15109573 TI - Beta-2-glycoprotein I is growth regulated and plays a role as survival factor for hepatocytes. AB - Beta-2-glycoprotein I (beta(2)GPI) is mainly produced by the liver and is found in plasma partially associated to lipoproteins. Although various properties have been attributed to this protein, its physiological role remains still unclear. We investigated its expression in cultured liver cells and in regenerating liver. Expression studies in HepG2 cells demonstrate that beta(2)GPI mRNA is regulated in a cell cycle-dependent manner, with very low expression in low cycling conditions and increasing levels in proliferating cells. p21 WAF-dependent growth arrest, induced by butyrate treatment, down-regulate beta(2)GPI mRNA levels. Immunolocalization in normal rat liver shows a non-homogeneous pattern, being mainly present in the centrolobular area; post-hepatectomy regenerating rat liver is uniformly immunostained and mitotic elements show the highest protein expression. Albumin gene expression, studies as control liver specific product, was not affected by sodium butyrate induced growth arrest. As previously reported for endothelial cells, beta(2)GPI behaves as survival factor for HepG2 cells: when increasing amounts of the protein (10-50 microg) have been added to serum deficient cultured liver cells a progressive reduced cell loss was observed. In conclusion, the present data demonstrate that beta(2)GPI gene expression is strictly related to the proliferative status of hepatic cells and that this protein could play a role in maintaining liver cells vitality when exposed to different stress factors such as regeneration after partial hepatectomy or growth factors depletion. PMID- 15109574 TI - Vigilin is co-localized with 80S ribosomes and binds to the ribosomal complex through its C-terminal domain. AB - The biological relevance of vigilin a ubiquitous multi (KH)-domain protein is still barely understood. Investigations over the last years, however, provided evidence for a possible involvement of vigilin in the nucleo-cytoplasmic transport of tRNA and in the subsequent association of tRNA with ribosomes. We therefore investigated the potential association of vigilin with 80S ribosomes. Immunostaining, gel filtration, westernblot analysis of polyribosomes and high salt treatment of 80S ribosomes isolated from fresh human placenta were applied to analyze the possible association of vigilin with ribosomes. Overlay assays were performed to examine whether vigilin is capable of binding to ribosomal proteins. Immunostaining of HEp-2 cells, gel filtration of a cytoplasmic extract of HEp-2 cells and westernblot analysis of isolated 80S ribosomes clearly demonstrate that vigilin is bond to the ribosomal complex. Vigilin detaches from the ribosomal complex under the influence of high salt concentrations. We present data that radioactively labeled human vigilin interacts directly with a subset of ribosomal proteins from both subunits. We were able to narrow down the putative binding region to the C-terminal domain by using vigilin mutant constructs. Therefore our results provide strong evidence that vigilin is bond to the ribosomal complex and underline the hypothesis that vigilin might be involved in the link between tRNA-export and the channeled tRNA-cycle on ribosomes. PMID- 15109575 TI - Cold-induced hyperthyroidism produces oxidative damage in rat tissues and increases susceptibility to oxidants. AB - In this work, we investigated whether cold exposure-induced hyperthyroidism increases oxidative damage and susceptibility to oxidants of rat liver, heart and skeletal muscle. All tissues exhibited gradual increases in hydroperoxide and protein-bound carbonyl levels. Glutathione peroxidase activity increased in all tissues after 2 days and further increased in the muscle after 10 days of cold exposure. Liver glutathione reductase activity increased after 10 days of cold exposure, while heart and muscle activities were not modified. Vitamin E levels were not affected by cold, while coenzyme Q9 and coenzyme Q10 levels decreased in heart and muscle after 2-day cold exposure and were not further modified after 10 days. Liver coenzyme Q9 levels increased after 2 days whereas coenzyme Q10 levels increased after 10 days in the cold. The whole antioxidant capacity was lowered, while parameters positively correlated with susceptibility to oxidants were increased by cold. Lipid fatty acid composition was modified in all tissues. In particular, fatty acid unsaturation degree increased in heart and muscle. Cytochrome oxidase activity increased, suggesting an increased content of hemoproteins, which are able to generate .OH radical. This view was supported by the observation that the tissue susceptibility to H(2)O(2) treatment, which is strongly correlated to iron-ligand content, increased after cold exposure. In this frame, it is apparent that the increase in oxidative capacity, necessary for homeotherm survival in low temperature environments, has potential harmful effects, because it results in increased susceptibility to oxidative challenge. PMID- 15109576 TI - Refolding intermediate of guanidine hydrochloride denatured aminoacylase. AB - The refolding of aminoacylase denatured in 6M guanidine hydrochloride (GdnHCl) has been studied by measuring enzyme activity, fluorescence emission spectra, ANS fluorescence spectra and far-UV circular dichroism spectra. The results showed that GdnHCl-denatured aminoacylase could be refolded and reactivated by dilution. A refolding intermediate was observed for low concentrations of GdnHCl (between 0.5 and 1.2M). This refolding intermediate was characterized by an increased fluorescence emission intensity, a blue-shifted emission maximum, and by increased binding of the fluorescence probe 8-anilino-1-naphthalenesulfonate (ANS). The secondary structure of the intermediate was similar to that of the native enzyme, and was therefore quite similar to the molten globule state often found in the protein folding pathway. Combined with the previous evidence of existence of an intermediate during unfolding process, we therefore proposed that the unfolding and refolding of aminoacylase might share the same pathway. A comparison of the Apo-enzyme and Holo-enzyme showed that there was little effect of the zinc ion on the refolding of the aminoacylase. Our study, the first successful report of the refolding of this metalloenzyme, also showed that lowering the concentration and the temperature of the enzyme improved the refolding rate of aminoacylase. The system therefore provides a useful model to study the refolding of proteins with prosthetic groups. PMID- 15109577 TI - Chaperone-like activity of mammalian elongation factor eEF1A: renaturation of aminoacyl-tRNA synthetases. AB - Eukaryotic translational elongation factor eEF1A is known to be responsible for the binding of codon-specific aminoacyl-tRNAs to the ribosome. In this study, we report that in addition to this canonical function, eEF1A is able to promote the renaturation of aminoacyl-tRNA synthetases (ARS) and protect them against denaturation by dilution. The full recovery of the phenylalanyl- (PheRS) and seryl-tRNA synthetase (SerRS) activities was achieved in the presence of 4 microM eEF1A, while bovine serum albumin at similar concentration had no renaturation effect. Remarkably, in vitro renaturation occurs at the molar ratio of eEF1A to ARS equivalent to that found in the cytoplasm of higher eukaryotic cells. The eEF1A.GDP and eEF1A.GTP complexes were shown to be similar in their effect on the phenylalanyl-tRNA synthetase renaturation. Thus, we conclude that the chaperone like activity of eEF1A might be important for maintaining the enzymes activity in the protein synthesis compartments of mammalian cells. PMID- 15109580 TI - Inflammation and neurodegeneration in Parkinson's disease. AB - The immunohistochemical demonstration of reactive microglia and activated complement components suggests that chronic inflammation occurs in affected brain regions in Parkinson's disease (PD). Evidence from humans and monkeys exposed to MPTP indicates this inflammation may persist many years after the initial stimulus has disappeared. Chronic inflammation can damage host cells. Reports in the literature indicate that antiinflammatory agents inhibit dopaminergic cell death in animal models of PD, and there is one epidemiological report that their use significantly diminishes the risk of PD in humans. There is a marked elevation in the mRNA levels for complement proteins and markers of activated microglia in affected regions in PD. The upregulation appears greater than that found in inflamed arthritic joints. These data support the hypothesis that chronic inflammation may play an important role, if secondary, in the pathogenesis of PD. PMID- 15109578 TI - Human recombinant thiamine triphosphatase: purification, secondary structure and catalytic properties. AB - Thiamine triphosphate (ThTP) is found in most living organisms and it may act as a phosphate donor for protein phosphorylation. We have recently cloned the cDNA coding for a highly specific mammalian 25 kDa thiamine triphosphatase (ThTPase; EC 3.6.1.28). As the enzyme has a high catalytic efficiency and no sequence homology with known phosphohydrolases, it was worth investigating its structure and catalytic properties. For this purpose, we expressed the untagged recombinant human ThTPase (hThTPase) in E. coli, produced the protein on a large scale and purified it to homogeneity. Its kinetic properties were similar to those of the genuine human enzyme, indicating that the recombinant hThTPase is completely functional. Mg2+ ions were required for activity and Ca2+ inhibited the enzyme by competition with Mg2+. With ATP as substrate, the catalytic efficiency was 10(-4) fold lower than with ThTP, confirming the nearly absolute specificity of the 25 kDa ThTPase for ThTP. The activity was maximum at pH 8.5 and very low at pH 6.0. Zn2+ ions were inhibitory at micromolar concentrations at pH 8.0 but activated at pH 6.0. Kinetic analysis suggests an activator site for Mg2+ and a separate regulatory site for Zn2+. The effects of group-specific reagents such as Woodward's reagent K and diethylpyrocarbonate suggest that at least one carboxyl group in the active site is essential for catalysis, while a positively charged amino group may be involved in substrate binding. The secondary structure of the enzyme, as determined by Fourier-transform infrared spectroscopy, was predominantly beta-sheet and alpha-helix. PMID- 15109581 TI - Alpha-synuclein, Parkinson's disease, and Alzheimer's disease. AB - Alpha synuclein (alpha-SN) is a ubiquitous protein that is especially abundant in the brain and has been postulated to play a central role in the pathogenesis of Parkinson's disease (PD), Alzheimer's disease, and other neurodegenerative disorders. Here, we show that alpha-SN plays dual role of neuroprotection and neurotoxicity depending on its concentration or level of expression. In addition, our study shows that alpha-synuclein is differentially expressed in human peripheral blood mononuclear cells. PD patients expressed more alpha-synuclein than healthy controls. Thus, the alpha-synuclein expression in the peripheral immune system might be one of the primary causes of immune abnormalities in PD patients. PMID- 15109582 TI - Dementia with Lewy bodies and Parkinson's disease. AB - Lewy bodies (LB) in the central nervous system are associated with several different clinical syndromes including Parkinson's disease (PD) and dementia with Lewy bodies (DLB). Long term follow up of PD patients finds up to 78% eventually develop dementia, most of these patients exhibiting fluctuating cognition and visual hallucinations similar to DLB and with extensive cortical LB at autopsy. alpha-Synuclein positive, neuritic pathology, in the putamen of DLB and Parkinson's disease dementia (PDD), may contribute to postural-instability gait difficulty, parkinsonism, diminished levodopa responsiveness and increased neuroleptic sensitivity. Cognitive and neuropsychiatric symptoms improve with cholinesterase inhibitor treatment in both patient groups. DLB and PDD should be seen as different points on a spectrum of LB disease. Distinguishing them as separate disorders may be useful in clinical practice, but may be of limited value in terms of investigating and treating the underlying neurobiology. PMID- 15109583 TI - Caribbean parkinsonism and other atypical parkinsonian disorders. AB - Atypical parkinsonism (AP) is a term applied to disorders characterized by parkinsonism that evolves rapidly, with poor or transient response to levodopa, or has other associated features such as early falls and postural instability, early autonomic failure, supranuclear gaze palsy, pyramidal or cerebellar signs, alien hand syndrome or severe ideomotor apraxia. The most common AP are multiple system atrophy, progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). Other APs include Caribbean parkinsonism (CP) and parkinsonism-dementia complex of Guam (PDC). In this review we provide an update in etiology, neuropathology, diagnosis and treatment of atypical parkinsonian disorders associated with protein tau deposit, also known as tauopathies. PMID- 15109584 TI - Sleep and fatigue in Parkinson's disease. AB - Sleep disorders and fatigue are common problems in Parkinson's disease (PD). Although they frequently appear together, they are often distinct symptoms that must be understood separately. Fatigue has been reported to be the most bothersome aspect of PD in about one-third of patients, yet it is poorly understood and not clearly treatable. Sleep disorders, while more common, are less bothersome to the patients and often responsive to therapy. An overview of sleep disorders in PD and an approach to therapy will also be outlined. The little that is known about fatigue in PD will be reviewed. PMID- 15109585 TI - Treatment of depression in Parkinson's disease. AB - Depression is an important and common nonmotor feature of Parkinson's disease (PD) that is associated with significant disability and a negative impact on quality of life. The physician should remain vigilant for symptoms of depression as they may be mistaken for the progression of Parkinson's disease itself. Transient dysphoria that occurs during 'off' periods in fluctuating PD patients must be distinguished from true depression. Antidepressant therapy should be instituted if depression is interfering with the patient's daily function. The use of serotonin reuptake inhibitors and tricyclic antidepressants in the treatment of depression in PD is widespread in clinical practice. Dopamine agonists may be effective in the treatment of milder depression as well. Individual or family counseling may be helpful. In patients with severe depression who are refractory to antidepressant medications, a series of electroconvulsive treatments can be lifesaving. Nonconventional therapies such as transcranial magnetic stimulation are being investigated. PMID- 15109586 TI - Rehabilitative approaches to Parkinson's disease. AB - Many sources of disability impact on quality of life and some are likely to remain relatively refractory to pharmacological or surgical treatment. Rehabilitative approaches may help despite the paucity of randomized control studies. There is increasing evidence from neuroscience, reviewed here, that supports current rehabilitative approaches and most importantly, can be the basis for future rehabilitative approaches. Parkinson's disease is complex and the disabilities not absolute but relative. These disabilities can be affected by the environment or context in which the motor activities occur. These observations can be exploited and argue for the potential of rehabilitative approaches. PMID- 15109587 TI - Deep brain stimulation surgery for Parkinson's disease: mechanisms and consequences. AB - Despite the introduction of new medications, motor fluctuations and dyskinesias disable a significant proportion of Parkinson's disease patients. This has lead to renewed interest in stereotactic neurosurgery. A skilled team is needed to ensure that patient assessment and selection, operative technique, intraoperative monitoring, and post-operative management are optimised. High frequency stimulation has similar effects to ablative surgery, and is generally preferred. The clinical effects and possible mechanisms of action of deep brain stimulation of the subthalamic nucleus and globus pallidus are reviewed. PMID- 15109588 TI - Tick-borne rickettsioses in international travellers. AB - BACKGROUND: Tick-borne rickettsioses are of emerging importance in today's travel medicine but have until recently received little attention. We describe the current knowledge of tick-borne rickettsioses as they relate to international travel, their microbiological diagnosis, treatment, possible prevention, and future prospects. METHODS: Literature-based review and personal observations. RESULTS: During the last decade, some 400 cases of tick-borne rickettsioses have been reported in international travellers, the vast majority being African tick bite fever caused by Rickettsia africae and Mediterranean spotted fever caused by Rickettsia conorii. Only a minority of infected travellers can recall a preceding tick bite. Most patients present with a mild-to-moderately severe flu-like illness typically accompanied by a cutaneous rash and an inoculation eschar at the site of the tick bite, but potentially life-threatening disease with disseminated vaculitis is occasionally seen. Definite microbiological confirmation of tick-borne rickettsioses by isolation or antigen detection is only available at reference laboratories and diagnosis must in most cases rely on clinical and epidemiological data supported by serology. Doxycycline is the recommended treatment for tick-borne rickettsioses and prevention is based on personal protective measures against tick bites when travelling in endemic areas. CONCLUSION: Tick-borne rickettsiosis should be suspected in febrile returnees from endemic areas, especially in cases with skin eruptions. Travellers to endemic areas should be encouraged to use personal protective measures against tick bites. PMID- 15109589 TI - Antimicrobial resistance of Gram-negative bacilli isolates from inpatients and outpatients at Yaounde Central Hospital, Cameroon. AB - OBJECTIVE: To determine and compare antimicrobial susceptibility patterns of pathogenic bacteria from inpatients and outpatients at a university teaching hospital in Yaounde, Cameroon. METHODS: Gram-negative bacilli isolates (n = 522), obtained from a wide range of clinical specimens (urine, pus and blood) from inpatients and outpatients at Yaounde Central Hospital between March 1995 and April 1998, were evaluated for resistance to antibiotics (amoxicillin, amoxicillin/clavulanate, piperacillin, cefazolin, cefoxitin, cefotaxime, ceftazidime, aztreonam, imipenem, gentamicin, tobramicin, ofloxacin and trimethoprim/sulfamethoxazole). RESULTS: Of the 522 isolates recorded, 80.3% were Enterobacteriaceae. A high incidence of resistance to amoxicillin (85%), piperacillin (75%) and trimethoprim/sulfamethoxazole (71%) was observed. The proportion of antimicrobial-resistant isolates from inpatients was significantly higher than that from outpatients (P < 0.05), except for piperacillin, tobramicin and trimethoprim/sulfamethoxazole. The combinations of antimicrobial and organism showed that the percentage of ceftazidime-resistant Pseudomonas aeruginosa and ceftazidime-resistant Enterobacter cloacae were 26.8% and 24% respectively. The rate of antimicrobial resistance in isolates from inpatients was not significantly higher than that in isolates from outpatients for all the antimicrobial/organism combinations, except for ceftazidime-resistant Escherichia coli, which was exclusively found in isolates from inpatients. Among Enterobacteriaceae, high and low level penicillinase (mostly in E. coli (13.6% and 11% respectively) and Klebsiella spp. (9% and 8% respectively) were the most important beta-lactam resistance phenotypes (31.2% and 23.6%, respectively). Wild type (exclusively observed in E. coli, Proteus mirabilis and Salmonella spp.) and low level penicillinase were higher in outpatient than inpatient isolates (wild type--17.9% vs 10.8% and low level penicillinase--29.4% vs 20.5%, respectively; P < 0.05). However, extended spectrum beta-lactamase strains (Klebsiella spp. (3.5%), E. coli (2.6%), Citrobacter spp. (0.7%), Enterobacter spp. (0.4%) and P. mirabilis (0.2%)) were exclusively recovered from inpatients. Penicillinase and high level cephalosporinase resistance phenotypes were frequently observed in non fermenter Gram-negative bacilli (46.6% and 29.1% respectively). However, there were no significant differences in penicillinase and cephalosporinase resistance between inpatient and outpatient isolates. CONCLUSION: As the incidence of antimicrobial resistance is substantially higher in isolates from inpatient than outpatient pathogens, more resources should be allocated within the hospital to encourage good antibiotic practices and good hospital hygiene. PMID- 15109590 TI - Infective complications according to duration of antibiotic treatment in acute abdomen. AB - INTRODUCTION: Adjuvant antibiotic therapy for acute abdominal conditions is widely used. Its timing, duration, dose and spectrum, however, are not homogeneous amongst surgeons and prolonged courses are often used despite the unproven benefits of this practice. OBJECTIVE: To evaluate use and compare duration of antibiotic treatments in acute abdominal surgery. METHODS: Retrospective cohort study. The medical records of 290 patients who underwent operations for acute abdomen from July 1998 to July 1999 in a teaching hospital were reviewed. The pattern of antibiotic use and rates of postoperative complications were evaluated, along with surgical diagnosis, degree of contamination/infection, and incidence of postoperative complications. The patients were stratified according to the degree of contamination/infection noted during the operation. The study population was divided in two groups according to the duration of antibiotic use (cut-off point at the median antibiotic use in days, for each group of contamination/infection degree), and outcomes were compared. RESULTS: The degree of contamination/infection was significantly associated with an increased risk of wound infection, intra-abdominal abscess, postoperative infective complications and overall postoperative complications (p < 0.001). A long course of antibiotics was not associated with lower infective complication rates. CONCLUSIONS: Shorter courses of antibiotic therapy based on the degree of contamination/infection seem to be safe. A prospective study should confirm this hypothesis. PMID- 15109591 TI - Infective endocarditis: a five-year experience at a tertiary care hospital in Pakistan. AB - OBJECTIVES: Infective endocarditis is common and data regarding its pattern and outcome from developing countries is sparse. We therefore examined the spectrum, demographics and clinical features of infective endocarditis and sought to determine the factors affecting its clinical outcome. PATIENTS AND METHODS: Over a five-year period at our university hospital in Karachi, Pakistan, we identified 66 patients with infective endocarditis and compared their clinicoepidemiologic features and outcomes to subjects in the West. RESULTS: The male:female ratio was 2:1; overall median age was 24 years (35.5 years for men and 13.5 years for women) (p < 0.001). Median duration of symptoms before presentation was 20.5 days. Major predisposing cardiac abnormalities included congenital (50%) and rheumatic (23%) lesions, and a history of heart surgery (17%). Causative organisms and valvular sites of infection were similar to those seen in developed countries. Blood culture-negative infective endocarditis was found in 48% of cases. Renal failure was more frequent among culture-positive patients (p = 0.055). Risk factors for mortality included neurologic (p = 0.003) and embolic (p = 0.02) complications, renal (p = 0.03) and left ventricular failure (p = 0.002), and a history of cardiac surgery (p = 0.026). Overall mortality was 27%. CONCLUSIONS: Patients with infective endocarditis in Pakistan exhibit significant differences compared to their counterparts in the West, including younger age at presentation, incidence of predisposing cardiac conditions, and gender differences reflecting sociocultural bias. Neurologic and embolic complications, renal failure and heart failure predict a worse outcome. PMID- 15109592 TI - A case of Pasteurella multocida peritoneal dialysis-associated peritonitis and review of the literature. AB - OBJECTIVES: Two episodes of peritoneal dialysis-associated peritonitis, which occurred four months apart and were both due to Pasteurella multocida, were noted in a 73 year old woman. This report aims to describe the clinical history of these episodes and the microbiological investigations that were undertaken. The relevant literature will also be discussed. METHODS AND RESULTS: Basic microbiological tests identified the organism as Pasteurella multocida, and further work at a specialist laboratory classified it as Pasteurella multocida subsp. multocida. Pulsed field gel electrophoresis confirmed that the strains isolated from the two clinical episodes originated from the same clone. A literature search was performed, looking particularly for patients who experienced more than one episode of peritonitis caused by Pasteurella spp, whether due to recurrence or re-infection. CONCLUSIONS: It is likely that the infection resulted from a domestic cat, as there was evidence of a cat bite to the dialysis tubing in the period between the two episodes. Re-infection with two identical strains of pasteurella is more probable than relapse, for reasons discussed. Strict hygiene and avoiding contact between dialysis tubing and domestic animals must be emphasised to try to prevent pasteurella and other animal-associated infections in this already vulnerable population. PMID- 15109593 TI - Simulated post-exposure rabies vaccination with purified chick embryo cell vaccine using a modified Thai Red Cross regimen. AB - OBJECTIVES: Currently, two intradermal regimens for the administration of cell culture rabies vaccines are approved by the WHO for rabies post-exposure prophylaxis: the two site Thai Red Cross regimen (TRC) and the eight site regimen. For the TRC regimen the volume of vaccine recommended per dose is 0.1 ml of purified Vero cell rabies vaccine (PVRV) and 0.2 ml of purified chick embryo cell vaccine (PCEC). The objective of the present study was to evaluate comparatively the immune response to PCEC and PVRV vaccines administered by the TRC regimen using a uniform dose of 0.1 ml of vaccine. METHODS: Forty-two subjects received TRC regimen (2-2-2-0-1-1) with 0.1 ml of PCEC vaccine and 38 subjects received the same regimen with PVRV. The rabies neutralizing antibody response in these subjects on days 10, 28, 90 and 180 was determined by the standard mouse neutralization test (MNT). RESULTS: There was adequate antibody response with both the vaccines and 100% seroconversion was observed by day 10. Furthermore, the antibody titers obtained with PCEC did not differ significantly from those obtained with PVRV on all days tested (p > 0.05). CONCLUSIONS: It can be concluded from the results that an adequate antibody response can be obtained with PCEC vaccine when administered by the TRC regimen even after reducing the quantity of vaccine from 0.2 ml to 0.1 ml per intradermal dose. The feasibility of using this regimen in true post-exposure cases needs to be further evaluated. PMID- 15109594 TI - The predictors of outcome in immunocompetent patients with hematogenous candidiasis. AB - OBJECTIVE: Clinical parameters that predict outcome in non-immunosuppressed candidemic patients are not fully understood. METHODS: Eighty-one consecutive episodes of candidemia were retrospectively evaluated in 75 patients during 1998 2000. RESULTS: Infection due to Candida albicans was common (n = 30; 37%) followed by Candida glabrata (n = 25; 31%), Candida parapsilosis (n = 14; 17%), Candida tropicalis (n = 6; 7%), Candida krusei (n = 5; 6%), and Candida lusitaniae (n = 1; 1%). Among 70 evaluable patients, 31 (44%) had fungemia associated mortality; advanced age (P < 0.004), underlying malignancy (P < 0.025), coronary artery disease (P < 0.01), and concurrent non-Candida species fungal infection (P < 0.047) were significant prognosticators of compromised short-term survival by multivariate analysis. Mortality was higher in patients with Candida glabrata (60%) and C. tropicalis (75%) infection compared to 44% deaths in individuals with C. albicans infection (P > 0.1). 11/25 (44%) of non immunocompromised individuals died and 20/45 (44%) immunosuppressed patients succumbed to fungemia: persistent vs. non-persistent (< 3 days) Candida bloodstream invasion, neutropenia, diabetes mellitus, renal insufficiency, prior antimicrobial therapy, cirrhosis of liver, abdomino-pelvis surgery, and critical care-unit vs. non critical-care-unit admission did not significantly impact outcome in either group. All 11 infants, including nine with prematurity, survived Candida species bloodstream infection (P < 0.025). CONCLUSIONS: Short term mortality in candidemic non-immunocompromised patients was comparable to fungemia-associated deaths in immunosuppressed patients. Ischemic heart disease has appeared as a new predictor of unfavorable outcome in patients with hematogenous candidiasis. PMID- 15109595 TI - Fluoroquinolone resistance trends and animal drug use: a retrospective analysis. PMID- 15109597 TI - Frontal brain abscess due to Streptococcus pneumoniae associated with an osteoma. PMID- 15109598 TI - Streptococcus pyogenes bacteremia in the setting of cholangitis. PMID- 15109600 TI - Depth-resolved structural imaging by third-harmonic generation microscopy. AB - Third harmonic generation microscopy is shown to be a robust method for obtaining structural information on a variety of biological specimens. Its nature allows depth-resolved imaging of inhomogeneities with virtually no background from surrounding homogeneous media. With an appropriate illumination geometry, third harmonic generation microscopy is shown to be particularly suitable for imaging of biogenic crystals, enabling extraction of the crystal orientation. PMID- 15109601 TI - Similar diffusibility of membrane proteins across the axon-soma and dendrite-soma boundaries revealed by a novel FRAP technique. AB - In polarized mature neurons, the asymmetrical distribution of proteins between axonal and somatodendritic plasma membrane (PM) domains may be maintained by a diffusion barrier at the axon-soma boundary. At the boundary, a complex containing membrane-associated and cytoskeletal proteins is formed, anchoring axonal membrane proteins and indirectly hindering the diffusion of other membrane proteins. We examined the latter case, i.e., secondary diffusion impedance by comparing the mobility of fluorescently labeled membrane proteins within the axon soma and dendrite-soma boundaries. We performed fluorescence recovery after photobleaching (FRAP) experiments using mature cultured hippocampal neurons that had been labeled specifically at their PMs with fluorescent proteins (FPs). The maturation of these neurons was confirmed by immunolocalization with Ankyrin-G, which is thought to participate in the creation of the diffusion barrier at the axon-soma boundary. We developed a wide-field microscope equipped with a device (digital micromirror device) composed of 1024 x 768 binary mirrors at the field stop, allowing free control of the illumination area and intensity. After the FPs in peripheral processes were photobleached, nonbleached FPs diffused into all the processes at equivalent speeds. These results indicate that the secondary diffusion barrier to exogenously overexpressed membrane proteins is not specific to the axon-soma boundary. PMID- 15109602 TI - Higher harmonic generation microscopy for developmental biology. AB - Optical higher harmonic generation, including second harmonic generation and third harmonic generation, leaves no energy deposition to its interacted matters due to an energy-conservation characteristic, providing the "noninvasiveness" nature desirable for biological studies. Combined with its nonlinearity, higher harmonic generation microscopy provides excellent three-dimensional (3D) sectioning capability, offering new insights into the studies of embryonic morphological changes and complex developmental processes. By choosing a laser working in the biological penetration window, here we present a noninvasive in vivo light microscopy with sub-micron 3D resolution and millimeter penetration, utilizing endogenous higher harmonic generation signals in live specimens. Noninvasive imaging was performed in live zebrafish (Danio rerio) embryos. The complex developmental processes within > 1-mm-thick zebrafish embryos can be observed in vivo without any treatment. No optical damage was found even with high illumination after long-term observations and the examined embryos all developed normally at least to the larval stage. The excellent 3D resolution of the demonstrated technology allows us to capture the subtle developmental information on the cellular or sub-cellular levels occurring deep inside the live embryos and larvae. This technique can not only provide in vivo observation of the cytoarchitecture dynamics during embryogenesis with submicron resolution and millimeter penetration depth, but would also make strong impact in developmental and structural biology studies. PMID- 15109603 TI - Dynamic interaction between BAF and emerin revealed by FRAP, FLIP, and FRET analyses in living HeLa cells. AB - Barrier-to-autointegration factor (BAF) is a conserved 10 kDa DNA-binding protein. BAF interacts with LEM-domain proteins including emerin, LAP2 beta, and MAN1 in the inner nuclear membrane. Using fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP), we compared the mobility of BAF to its partners emerin, LAP2 beta, and MAN1 in living HeLa cells. Like endogenous BAF, GFP-BAF was enriched at the nuclear envelope, and found inside the nucleus and in the cytoplasm during interphase. At every location, FRAP and FLIP analysis showed that GFP-BAF diffused rapidly; the halftimes for recovery in a 0.8 microm square area were 260 ms at the nuclear envelope, and even faster inside the nucleus and in the cytoplasm. GFP-fused emerin, LAP2 beta, and MAN1 were all relatively immobile, with recovery halftimes of about 1 min, for a 2 microm square area. Thus, BAF is dynamic and mobile during interphase, in stark contrast to its nuclear envelope partners. FLIP results further showed that rapidly diffusing cytoplasmic and nuclear pools of GFP-BAF were distinctly regulated, with nuclear GFP-BAF unable to replenish cytoplasmic BAF. Fluorescence resonance energy transfer (FRET) results showed that CFP-BAF binds directly to YFP-emerin at the inner nuclear membrane of living cells. We propose a "touch-and-go" model in which BAF binds emerin frequently but transiently during interphase. These findings contrast with the slow mobility of both GFP-BAF and GFP-emerin during telophase, when they colocalized at the 'core' region of telophase chromosomes at early stages of nuclear assembly. PMID- 15109604 TI - Quantitative characterization of aqueous solutions probed by the third-harmonic generation microscopy. AB - Third-harmonic microscopy is one of the emerging techniques for noninvasive microscopic imaging of biological structures. We use a novel technique for nonlinear optical material characterization and study the effect of different environment and the structural sensitivity of the third harmonic. In particular, a transformation of collagen in solution is observed for the first time using third-harmonic generation. We also study the ultimate limits of the third harmonic to detect micro- and nanoscopic features inside living cells and find that structures as small as 50 nm can be detected using the current level of technology. PMID- 15109605 TI - The androgen receptor ligand-binding domain stabilizes DNA binding in living cells. AB - The androgen receptor (AR) is a member of the steroid receptor family, a group of transcription factors that activate steroid-regulated genes. Live cell studies of several steroid receptors have shown that the mobility of the liganded receptor is strongly reduced compared to the unliganded receptor. To investigate the nature of this reduced mobility, we generated Hep3B cells stably expressing green fluorescent protein (GFP)-AR at physiological levels. Computer-aided analysis of photobleaching experiments showed that in the presence of ligand on average one out of five ARs is immobilized, each individual AR being immobile for 1-2 min. This immobilization depended on DNA binding since GFP-ARs mutated in the DNA binding domain were not immobilized. Interestingly, a truncated AR lacking the ligand-binding domain (LBD) displayed substantially shorter immobilizations, in the order of seconds, although its transcriptional activation function was stronger. Our data suggest the LBD has a role in maintaining the stability of AR DNA complexes. PMID- 15109606 TI - Red-edge anisotropy microscopy enables dynamic imaging of homo-FRET between green fluorescent proteins in cells. AB - Steady-state fluorescence anisotropy measurements can be used to detect fluorescence resonance energy transfer (FRET) between identical fluorophores (homo-FRET). However, the contribution of homo-FRET to the steady-state anisotropy must be discerned from those due to the orientational distribution and rotational diffusion, which so far has required photobleaching controls, largely precluding dynamic measurements in live cells. We describe a variation of steady state anisotropy microscopy in which the contribution of homo-FRET is dynamically isolated from the total anisotropy by exploiting the loss of energy transfer that occurs at red-edge excitation. Excitation of enhanced green fluorescent protein (EGFP) at the red-edge of its absorption band shows the shift in the emission spectrum compared to main-band excitation that is characteristic for photo selection of static low energy S(0)-S(1) transitions that fail to exhibit FRET. An experimental setup for steady-state fluorescent anisotropy microscopy is described that can be used to acquire anisotropy images in live cells at main band and red-edge excitation of EGFP. We demonstrate in live cells homo-FRET suppression of protein fusion constructs that consist of two and three EGFP molecules connected by short linkers. This methodology represents a novel approach for the dynamic measurement of homo-FRET in live cells that will be of utility in the biological sciences to detect oligomerization and concentration dependent interactions between identically labeled molecules. PMID- 15109607 TI - 4Pi-microscopy of the Golgi apparatus in live mammalian cells. AB - We report the applicability of 4Pi-microscopy to live mammalian cells. Controlled interference of the counterpropagating wavefronts is possible despite the slight variations in cellular refractive index. Superresolved 3D-fluorescence imaging is exemplified with the first representation of the Golgi apparatus in a live cell at approximately 100 nm resolution. PMID- 15109608 TI - Lipid domain formation and dynamics in giant unilamellar vesicles explored by fluorescence correlation spectroscopy. AB - Lipids in eukaryotic cell membranes have been shown to cluster in "rafts" with different lipid/protein compositions and molecular packing. Model membranes such as giant unilamellar vesicles (GUVs) provide a key system to elucidate the physical mechanisms of raft assembly. Despite the large amount of work devoted to the detection and characterization of rafts, one of the most important pieces of information still missing in the picture of the cell membrane is dynamics: how lipids organize and move in rafts and how they modulate membrane fluidity. This missing element is of crucial importance for the trafficking at and from the periphery of the cell regulated by endo- and exocytosis and, in general, for the constant turnover which redistributes membrane components. Here, we review studies of combined confocal fluorescence microscopy and fluorescence correlation spectroscopy on lipid dynamics and organization in rafts assembled in GUVs prepared from various lipid mixtures which are relevant to the problem of raft formation. PMID- 15109609 TI - Hormones and the cardiovascular system. PMID- 15109610 TI - Is aldosterone bad for the heart? PMID- 15109611 TI - Aldosterone and DNA: the 50th anniversary. PMID- 15109612 TI - Mechanisms of mineralocorticoid action: determinants of receptor specificity and actions of regulated gene products. AB - The mineralocorticoid receptor (MR) and its close cousin, the glucocorticoid receptor (GR), share considerable structural and functional similarity, including indistinguishable DNA binding properties, yet they mediate distinct physiological responses in some tissues. Specificity is determined by their distinct interactions with other protein factors and modification by peptides, including the small ubiquitin modifier SUMO1. Serum and glucocorticoid-regulated kinase 1 (sgk1) is one key target gene of both MR and GR, and encodes a serine-threonine kinase that stimulates the apical membrane localization of the epithelial sodium channel ENaC. Sgk1 exerts its effects, at least in part, by inhibiting an isoform of the ENaC inhibitory ubiquitin ligase Nedd4-2. This review briefly summarizes two areas of mineralocorticoid research: molecular determinants of MR specificity, and the role of Sgk1 in mediating the effects of aldosterone on epithelial Na(+) transport. PMID- 15109613 TI - Selective activation of thyroid hormone signaling pathways by GC-1: a new approach to controlling cholesterol and body weight. AB - The current report describes progress in development of a selective thyroid hormone receptor modulator, GC-1. This compound binds selectively to the beta isoform of the thyroid hormone receptor, and its uptake into the heart is relatively low. Studies in rats, mice and monkeys show that GC-1 lowers cholesterol with 600- to 1400-fold more potency and approximately two- to threefold more efficacy than atorvastatin, a compound that blocks HMG-CoA reductase. GC-1 also decreases plasma levels of triglyceride and lipoprotein (a), and induces loss of fat. These effects can be observed under conditions where there is either no or minimal effect on heart rate, and no detectable loss of muscle. Although more study is required, compounds of this class deserve further investigation for treating lipid disorders and obesity. PMID- 15109614 TI - PPARs and LXRs: atherosclerosis goes nuclear. AB - Atherosclerosis is the leading cause of mortality in the Western world, and new therapeutics to target the metabolic and inflammatory factors that underlie its pathogenesis are needed. Peroxisome proliferator-activated receptors and liver X receptors are lipid-activated nuclear receptors that regulate systemic glucose and lipid metabolism, and modulate inflammation within the vascular wall. New understanding of their functions in physiology and the development of high affinity synthetic ligands highlight their potential as targets for the treatment of cardiovascular disease. PMID- 15109615 TI - ACE2, a new regulator of the renin-angiotensin system. AB - Angiotensin-converting enzyme (ACE) is a zinc metalloproteinase and a key regulator of the renin-angiotensin system (RAS). ACE2 is a newly described enzyme identified in rodents and humans with a more restricted distribution than ACE, and is found mainly in heart and kidney. ACE2 cleaves a single residue from angiotensin I (Ang I) to generate Ang 1-9, and degrades Ang II, the main effector of the RAS, to the vasodilator Ang 1-7. The importance of ACE2 in normal physiology and pathophysiological states is largely unknown. ACE2 might act in a counter-regulatory manner to ACE, modulating the balance between vasoconstrictors and vasodilators within the heart and kidney, and playing a significant role in regulating cardiovascular and renal function. PMID- 15109616 TI - Clinical applications of B-type natriuretic peptides. AB - Diagnostic, prognostic and therapeutic applications of B-type natriuretic peptides (NPs) will probably become part of routine management of heart failure within five years. Cardiac release of NPs rises with increasing cardiac dysfunction. Their secretion and plasma levels respond to intracardiac distending pressures, with other modulating influences including age, sex, renal function and other aspects of neurohormonal status. Single and serial plasma NP measurements, particularly of B-type and N-terminal pro-B-type NP, show promise in diagnosis of heart failure, risk stratification in those with known heart disease, and in adjustment of anti-failure therapy. Recombinant B-type NP is an effective parenteral treatment in decompensated heart failure. These applications of B-type NPs require confirmation before they become established in routine management of heart failure. PMID- 15109617 TI - Urotensin II: the old kid in town. AB - Urotensin II (U-II) is a vasoactive hormone that acts through a recently described seven transmembrane-spanning G-protein-coupled receptor called GPR14. Although touted as the most potent vasoconstrictor peptide yet identified, the responses elicited by U-II are species-, tissue- and endothelium-dependent. Available data question the contribution of U-II to resting cardiovascular homeostasis in humans; instead they point to a role for this hormone in disease (heart failure and cardiac cell growth, renal function, diabetes, and mitogenesis in vascular and tumour cells). Key features of these diseases are increased expression and activity of U-II receptors. In this review, we focus on recent evidence that supports a role of U-II and its receptor in cardiovascular disease. PMID- 15109618 TI - The developmental origins of the metabolic syndrome. AB - Both epidemiological and clinical evidence suggest relationships between the antenatal environment and the risk of developing insulin resistance and associated cardiovascular disease (part of the metabolic syndrome) in middle age. However, interpretation of these findings has been controversial. Recent experimental observations provide considerable evidence for a causal model linking adaptive responses to early environmental cues and the later risk of disease. Evolutionary and life history theory provide possible explanations of why these phenomena have persisted and how they might cause disease. In this article, we review the clinical and experimental perspectives on the "developmental origins of disease" model in the context of these new concepts. PMID- 15109619 TI - Synthesis of [O-methyl-11C]1-(2-chlorophenyl)-5-(4-methoxyphenyl)-4-methyl-1H pyrazole-3-carboxylic acid piperidin-1-ylamide: a potential PET ligand for CB1 receptors. AB - Synthesis and in vitro evaluation of [O-methyl-(11)C]1-(2-chlorophenyl)-5-(4 methoxyphenyl)-4-methyl-1H-pyrazole-3-carboxylic acid piperidin-1-ylamide ([(11)C]-1), a potential imaging agent for CB(1) receptors using PET is described. 1-(2-Chlorophenyl)-5-(4-hydroxyphenyl)-4-methyl-1H-pyrazole-3 carboxylic acid piperidin-1-ylamide (5), the precursor for radiolabeling, was synthesized from 4-OTBDPS-propiophenone (2) in five steps with 30% overall yield. The reaction of alcohol 5 with [(11)C]MeOTf at 60 degrees C afforded [(11)C]-1 with an average radiochemical yield of 14.5% (EOS) and >2000 Ci/mmol specific activity. The radiotracer was found to selectively label CB(1) receptors in slide mounted sections of postmortem human brain containing prefrontal cortex as demonstrated by in vitro autoradiography using phosphor imaging. PMID- 15109620 TI - Bicyclic anti-VZV nucleosides: thieno analogues bearing an alkylphenyl side chain have reduced antiviral activity. AB - Thieno analogues of the potent and selective furo-pyrimidine anti-VZV nucleoside family bearing a p-alkylphenyl side chain have been synthesised and tested for their antiviral activity against Varicella-Zoster virus (VZV). While the alkyl chain analogues were shown to retain full antiviral activity against VZV, these new analogues did not when compared to their furo parent nucleosides. PMID- 15109621 TI - Synthesis and biological evaluation of novel 2-pyridinyl-[1,2,3]triazoles as inhibitors of transforming growth factor beta 1 type 1 receptor. AB - A series of 2-pyridinyl-[1,2,3]triazoles have been synthesized and evaluated for their ALK5 inhibitory activity in the luciferase reporter assays. Compound 8d showed significant ALK5 inhibition (SBE-luciferase activity, 25%; p3TP-luciferase activity, 17%) at a concentration of 5 microM that is comparable to that of SB 431542 (SBE-luciferase activity, 21%; p3TP-luciferase activity, 12%), but weak p38 alpha MAP kinase inhibition (13%) at a concentration of 10 microM that is much lower than that of SB-431542 (54%). PMID- 15109622 TI - Site-specific conjugation of oligonucleotides to the C-terminus of recombinant protein by expressed protein ligation. AB - We developed a convenient method for synthesizing homogeneous DNA-protein conjugates. The method is based on expressed protein ligation of intein-fusion proteins and oligonucleotides derivatized with a cysteine. A range of cysteinyl oligonucleotides were synthesized by using a new reagent 1 and were successfully applied to expressed protein ligation to attach the oligonucleotides specifically at the C-terminus of a recombinant protein. PMID- 15109623 TI - Discovery and structure-activity relationship of coumarin derivatives as TNF alpha inhibitors. AB - The discovery and structure-activity relationship of a novel series of coumarin based TNF-alpha inhibitors is described. Starting from the initial lead 1a, various derivatives were prepared surrounding the coumarin core structure to optimize the in vitro inhibitory activity of TNF-alpha production by human peripheral blood mononuclear cells (hPBMC), stimulated by bacterial lipopolysaccharide (LPS). Selected compounds also demonstrated in vivo inhibition of TNF-alpha production in rats. PMID- 15109624 TI - Synthesis and antibacterial activity of 6-O-arylpropargyl-9-oxime-11,12-carbamate ketolides. AB - A series of novel 6-O-arylpropargyl-9-oxime-ketolides was synthesized and evaluated against various pathogens. These new compounds show promising in vitro antibacterial potency and in vivo efficacy against macrolide resistant strains. PMID- 15109625 TI - Synthesis of benzophenone oxime analogues as inhibitor of secretory phospholipase A(2) with anti-inflammatory activity. AB - The title compound have been synthesized and tested for structure activity relationship for Phospholipase A(2) (PLA(2)) [E.C. 3.1.1.4] enzyme inhibition. The in vitro PLA(2) enzyme inhibitory activity of benzophenone oxime analogue and in vivo anti-inflammatory activity studies using mice are highlighted. PMID- 15109626 TI - Synthesis and histone deacetylase inhibitory activity of cyclic tetrapeptides containing a retrohydroxamate as zinc ligand. AB - Cyclic tetrapeptide retrohydroxamic acids were prepared as histone deacetylase (HDAC) inhibitors and evaluated the inhibitory activity and found that they have potential as anticancer drugs. PMID- 15109627 TI - Potentiation of cytotoxic drug activity in human tumour cell lines, by amine substituted 2-arylbenzimidazole-4-carboxamide PARP-1 inhibitors. AB - The synthesis and biological evaluation of a new series of amine-substituted 2 arylbenzimidazole-4-carboxamide inhibitors of the DNA-repair enzyme poly(ADP ribose) polymerase-1 (PARP-1) is reported. The introduction of an amine substituent at the 2-aryl position is not detrimental to activity, with most inhibitors exhibiting K(i) values for PARP-1 inhibition in the low nanomolar range. Two compounds in this series were found to potentiate the cytotoxicity of the DNA-methylating agent temozolomide by 4-5-fold in a human colorectal cancer cell line. PMID- 15109628 TI - Supported synthesis of ferrocene modified oligonucleotides as new electroactive DNA probes. AB - The use of 1-[3-O-(2-cyanoethyl-N,N-diisopropylphosphor amidityl)propyl]ferrocene and 1-[3-O-dimethoxytrityl propyl]-1'-[3'-O-(2-cyanoethyl-N,N diisopropylphosphoramidityl) propyl] ferrocene as reactive synthons for DNA/RNA synthesizer allows to generate ferrocene-labelled oligonucleotides with remarkable DNA detection properties. PMID- 15109629 TI - Facile synthesis of fused 1,2,4-triazolo[1,5-c]pyrimidine derivatives as human adenosine A3 receptor ligands. AB - A facile synthetic method for fused triazolopyrimidine derivatives having high affinity and selectivity for human adenosine A(3) receptors is reported. The fused triazolopyrimidine derivatives were easily prepared by one-pot reaction using acylhydrazines and imidates prepared from amine derivatives bearing cyano group and orthoesters in situ. This synthetic method was useful in finding new tricyclic adenosine A(3) receptor antagonists and also in diversifying the substituents at two positions on the fused triazolopyrimidine ring. PMID- 15109630 TI - Enzyme cleavable and biotinylated photoaffinity ligand with diazirine. AB - The efficient synthesis of an enzyme cleavable biotinylated diazirinyl photoaffinity ligand is described to allow the effective manipulation of the photolabeled biocomponents. The compound contains a glutamic acid gamma-methyl ester, which is a precursor of the substrate for V8 protease, between the diazirinyl photophor and biotin moiety. After alkaline hydrolysis of the ester, the compound can be proteolyzed at the Glu moiety by V8 protease. The photophore was introduced to L-Phe p-nitroanilide and the ligand was applied to photolabel of chymotrypsin to manipulate the application of the concept. PMID- 15109631 TI - Discovery and SAR of potent, orally available and brain-penetrable 5,6-dihydro-4H 3-thia-1-aza-benzo[e]azulen- and 4,5-dihydro-6-oxa-3-thia-1-aza-benzo[e]azulen derivatives as neuropeptide Y Y5 receptor antagonists. AB - Combination of structural elements from a potent Y5 antagonist (2) with thiazole fragments that exhibit weak Y5 affinities followed by lead optimisation led to the discovery of (5,6-dihydro-4H-3-thia-1-aza-benzo[e]azulen-2-yl)-piperidin-4 ylmethyl-amino and (4,5-dihydro-6-oxa-3-thia-1-aza-benzo[e]azulen-2-yl)-piperidin 4-ylmethyl-amino derivatives. Both classes of compounds are capable of delivering potent and selective orally and centrally bioavailable NPY Y5 receptor antagonists. PMID- 15109632 TI - Synthesis and characterization of a DNA analogue stabilized by mercapto C nucleoside induced disulfide bonding. AB - A redox-active nucleobase analogue of a nucleotide was synthesized and incorporated into DNA using phosphoramidite chemistry. An analogue-containing oligonucleotide in the absence of a reducing reagent formed a stable duplex with a substantially higher melting temperature compared to that of a standard DNA duplex of the same length. PMID- 15109633 TI - Identification and synthesis of [1,2,4]triazolo[3,4-a]phthalazine derivatives as high-affinity ligands to the alpha 2 delta-1 subunit of voltage gated calcium channel. AB - We have identified and synthesized a series of [1,2,4]triazolo[3,4-a]phthalazine derivatives as high-affinity ligands to alpha 2 delta-1 subunit of voltage gated calcium channels. Structure-activity relationship studies directed toward improving the potency and physical properties of 2 lead to the discovery of 20 (IC(50)=15 nM) and (S)-22 (IC(50)=30 nM). A potent and selective radioligand, [(3)H]-(S)-22 was also synthesized to demonstrate that this ligand binds to the same site as gabapentin. PMID- 15109634 TI - SAR development of a selective 5-HT1D antagonist/serotonin reuptake inhibitor lead using rapid parallel synthesis. AB - Incorporation of an SRI (serotonin reuptake inhibitor) pharmacophore into a selective 5-HT(1D) agonist has led to the discovery of a molecule having both 5 HT(1D) antagonist and SRI activity. RPS methodology was used to develop the SAR and identify potential approaches to reduce unwanted adrenergic alpha 1 and dopamine D(2) cross-reactivities. PMID- 15109635 TI - Linker-modified quinoline derivatives targeting HIV-1 integrase: synthesis and biological activity. AB - A novel series of HIV-1 integrase inhibitors was synthesized and tested in both in vitro and ex vivo assays. These inhibitors are featured by the presence of a quinoline subunit and an ancillary aromatic ring linked by functionalized spacers such as amide, hydrazide, urea and 1-hydroxyprop-1-en-3-one moiety. Amide derivatives are the most promising ones and could serve as leads for further developments. PMID- 15109636 TI - Stereodefined and polyunsaturated inhibitors of histone deacetylase based on (2E,4E)-5-arylpenta-2,4-dienoic acid hydroxyamides. AB - Syntheses of (2E,4E)-5-arylpenta-2,4-dienoic acid hydroxyamides are described, some of which are potent inhibitors of histone deacetylase, a double bond conferring more than a 10-fold increase in potency compared with the triple bond analogue oxamflatin. Variation of substituents on the aromatic ring has a marked effect on potency, in vitro IC(50) values down to 50 nM being obtained. PMID- 15109637 TI - Statin-derived 1,3-oxazinan-2-ones as submicromolar inhibitors of LFA-1/ICAM-1 interaction: stabilization of the metabolically labile vanillyl side chain. AB - Modification of the vanillyl substituent on a potent, semisynthetic lymphocyte function-associated antigen (LFA)-1/intercellular adhesion molecule (ICAM)-1 binding inhibitor of the statin family resulted in metabolically more stable analogues that displayed submicromolar inhibitory activity in vitro and considerable anti-inflammatory activity in vivo. The benzodioxole derivative 2b emerged with the best overall profile. PMID- 15109638 TI - Design and synthesis of bridged gamma-lactams as analogues of beta-lactam antibiotics. AB - Anti-Bredt bridged bicyclo[3.2.1] gamma-lactams were designed as inhibitors of penicillin binding proteins (PBPs). The compounds were prepared by a carbenoid insertion into a lactam N-H bond. Their weak antibacterial activity could either be explained by a poor chemical stability or by unfavorable steric interactions of the methylene bridge of the gamma-lactam with the targeted enzymes. PMID- 15109639 TI - MexAB-OprM specific efflux pump inhibitors in Pseudomonas aeruginosa. Part 4: Addressing the problem of poor stability due to photoisomerization of an acrylic acid moiety. AB - Exchange of the ethylene tether in a series of pyridopyrimidine-based MexAB-OprM specific efflux pump inhibitors to an amide bond stabilized the olefin of the acrylic acid moiety, preventing facile photoisomerization to the Z-isomer. Furthermore, the activity was drastically improved in the amide tether variants, providing extremely potent acrylic acid and vinyl tetrazole analogues. PMID- 15109640 TI - Syntheses of sphingosine-1-phosphate analogues and their interaction with EDG/S1P receptors. AB - Sphingosine-1-phosphate (S1P) is an important regulator of a wide variety of biological processes acting as an endogenous ligand to EDG/S1P receptors. In an effort to establish structure-activity relationship between EDG/S1P and ligands, we report herein homology modeling study of EDG-1/S1P(1), syntheses of S1P analogues, and cell based binding affinity study for EDG/S1P receptors. PMID- 15109641 TI - Rational design and synthesis of novel heparan sulfate mimetic compounds as antiadhesive agents. AB - A biological evaluation of the antiadhesive activity of novel heparan sulfate glycosaminoglycans mimetic compounds (KI-compounds) is described. In an adhesion assay, KI-111 [2-(4-fluoro-3-nitrobenzoyl)benzoic acetic anhydride] was found to exert potent inhibitory activities against the adhesion of human fibrosarcoma HT1080 cells and HeLa cells to fibronectin. Cell growth, migration, and invasion of HT1080 cells were also inhibited by KI-111 at almost equal concentrations. PMID- 15109642 TI - New pyrazolo[3,4-d]pyrimidines endowed with A431 antiproliferative activity and inhibitory properties of Src phosphorylation. AB - New 4-aminopyrazolo[3,4-d]pyrimidines bearing various substituents at the position 1 and 6, were synthesized. The new compounds showed antiproliferative activity toward A431 cells, were found to be inhibitors of Src phosphorylation, and induced apoptotic cell death. In particular, 2h was a better inhibitor of Src phosphorylation than the reference compound PP2. PMID- 15109643 TI - Design, synthesis, and structure-activity relationships of pyrazolo[3,4 d]pyrimidines: a novel class of potent enterovirus inhibitors. AB - A series of pyrazolo[3,4-d]pyrimidines were synthesized and their antiviral activity was evaluated in a plaque reduction assay. It is very interesting that this class of compounds provide remarkable evidence that they are very specific for human enteroviruses, in particular, coxsackieviruses. Some derivatives proved to be highly effective in inhibiting enterovirus replication at nanomolar concentrations. SAR studies revealed that the phenyl group at the N-1 position and the hydrophobic diarylmethyl group at the piperazine largely influenced the in vitro antienteroviral activity of this new class of potent antiviral agents. It was found that the pyrazolo[3,4-d]pyrimidines with a thiophene substituent, such as compounds 20-24, in general exhibited high activity against coxsackievirus B3 (IC(50) = 0.063-0.089 microM) and moderate activity against enterovirus 71 (IC(50) = 0.32-0.65 microM) with no apparent cytotoxic effect toward RD (rhabdomyosarcoma) cell lines (CC(50) > 25 microM). PMID- 15109644 TI - Structure of pentasaccharide of glycopeptide from TIME-EA4, N-glycoprotein in silkworm diapause eggs. AB - The TIME-EA4, from silkworm diapause eggs of pure strain C108, Bombyx mori, has glycosylated chain as tetrasaccharide (Man(2)GlcNAc(2)) attaching to the Asn(22) of T3 peptide from tryptic digests. On the other hand, from Showa silkworm strain we additionally observed a pentasaccharide (Man(3)GlcNAc(2)) on T3 at the same linkage site. The linkage pattern of the 5-sugar chain was studied through Smith degradation combined with LC-MS and MS/MS analyses. These advanced methods led us to conclude that the pentasaccharide was branching as Man 1-->3(Man 1-->6)Man 1- >4GlcNAc 1-->4GlcNAc. PMID- 15109645 TI - Synthesis, structural characterization, and antitumor activity of palladium(II) complexes containing a sugar unit. AB - Six palladium(II) complexes as cisplatin derivatives with a sugar unit (D glucose, D-galactose, D-mannose, D-xylose, and maltose) have been prepared. The structural features of the complexes have been characterized by NMR spectroscopy, elemental analysis, mass spectroscopy, and X-ray crystallography. The complexes have been tested for in vivo cytotoxicity against P388 cells implanted in mice. All of Pd compounds are apparently nontoxic. A T/C value of 120% was obtained for maltose derivative at the dose of 400 mg/kg, which indicates that the compound may be endowed with antitumor activity. PMID- 15109646 TI - Effect of gabapentin derivates on mechanical allodynia-like behaviour in a rat model of chronic sciatic constriction injury. AB - A series of mutual prodrugs derived from gabapentin, pregabalin, memantine, venlafaxine were synthesized and their pharmacological properties to treat neuropathic pain were investigated in a rat model of chronic sciatic nerve constriction injury (CCI). In vivo evaluation demonstrated that the mutual prodrugs 2002413A, 2002823A composed of two gabapentins, 2002414 composed of gabapentin and pregabalin were effective in reversal tactile allodynia in CCI rats. The prodrugs 2002413A, 2002414 had no significant influence on the rotarod activity. The result suggest that the prodrugs may be possible candidates for further development. PMID- 15109647 TI - Orally bioavailable small molecule ketoamide-based inhibitors of cathepsin K. AB - An orally available series of ketoamide-based inhibitors of cathepsin K has been identified. Starting from a potent inhibitor with poor oral bioavailability, modifications to P1 and P1' elements led to enhancements in solubility and permeability. These improvements resulted in orally available cathepsin K inhibitors. PMID- 15109648 TI - Structure-activity requirements for the antiproliferative effect of troglitazone derivatives mediated by depletion of intracellular calcium. AB - Depletion of calcium from the endoplasmic reticulum has shown to affect protein synthesis and cell proliferation. The anticancer effect of troglitazone was reported to be mediated by depletion of intracellular calcium stores resulting in inhibition of translation initiation. The unsaturated form of troglitazone displays similar anticancer properties in vitro. In this letter, we report our findings on the minimum structural requirements for both compounds to retain their calcium release and antiproliferative activities. PMID- 15109649 TI - Estrogen receptor ligands. Part 3: The SAR of dihydrobenzoxathiin SERMs. AB - A series of 3-alkyl, 3-cycloalkyl, and 3-heteroaryl dihydrobenzoxathiin analogs 1 were prepared and evaluated for estrogen/anti-estrogen activity in both in vitro and in vivo models. In general, the compounds were found to exhibit a high degree of selectivity for ER alpha over ER beta, but were less potent than the original lead compound 1a in the inhibition of estradiol-driven uterine proliferation. PMID- 15109650 TI - Synthesis and biological activity of macrocyclic taxane analogues. AB - A series of paclitaxel analogues possessing a macrocyclic structure between the 7 and 10 positions has been prepared. These compounds possess in vitro activity against a paclitaxel resistant cell line and have in vivo activity comparable to paclitaxel. PMID- 15109651 TI - Simple aromatic compounds containing propenone moiety show considerable dual COX/5-LOX inhibitory activities. AB - For the development of safer anti-inflammatory agents, simple aromatic compounds containing propenone moiety were prepared and evaluated for their dual COX/5-LOX inhibitory activities. Among the 17 prepared compounds, most of the compounds exhibited considerable COX/5-LOX inhibitory activities. Especially compound C(15) showed the most significant dual COX/5-LOX inhibitory activity. PMID- 15109652 TI - A novel 11-residual peptaibol-derived carrier peptide for in vitro oligodeoxynucleotide delivery into cell. AB - Using a pore- and channel-forming peptide, TV-XIIa, which is an 11-residual peptaibol isolated from the fungus Trichoderma viride, we developed a vehicle for the cellular delivery of such polar biologically active agents as antisense oligodeoxynucleotides (ODNs). To function as an ODN carrier, basic amino acids, 10-mer of lysine, were conjugated to the C-terminus of TV-XIIa and the designed carrier peptide, Ac-U-N-I-I-U-P-L-L-U-P-I-K-K-K-K-K-K-K-K-K-K-OH (U: alpha aminoisobutyric acid), was synthesized by the Fmoc-based solid-phase method. The complex between the carrier peptide and ODNs, which was electrostatically formed, was capable of crossing the membranes of NIH3T3 cells and the ODNs were accumulated in the cytoplasm and the nucleus. However, the complex was not taken up by A549 cells. The translocation of the complex occurred at both 4 and 37 degrees C in NIH3T3 cells and did not seem to involve an energy-dependent endocytic process. PMID- 15109653 TI - Discovery of a potent and selective alpha v beta 3 integrin antagonist with strong inhibitory activity against neointima formation in rat balloon injury model. AB - A new series of phenylpiperazine-based derivatives with strong antagonistic activity for alpha v beta 3 integrin were synthesized. Of these derivatives, the fluorine-substituted compound 8 showed strong inhibitory activity and high selectivity for alpha v beta 3 integrin receptor (IC(50) = 0.055 nM). In vivo evaluation of the antistenotic effects of 8 indicated that this compound significantly inhibits neointima formation in rat balloon injury model. PMID- 15109654 TI - Synthesis and glucose-6-phosphatase inhibitory activity of (thiouriedo)alkanoic acid esters. AB - A series of (3-pyridin-2-yl-thiouriedo)alkanoic acid esters (5a-j) have been synthesized by the reaction of pyridin-2-yl-dithiocarbamic acid methyl ester (2) and amino acid esters (4). Most of the synthesized compounds have been evaluated against glucose-6-phosphatase enzyme but only four compounds (5g-j) displayed significant inhibitory activity of the enzyme. PMID- 15109655 TI - Peripheral substituents of di(pyridiumyl)porphyrins affected on their interactions with DNA. AB - meso- and beta-Substituted di(pyridiumyl)porphyrins 3, 4, and 7 have been synthesized and their interactions with DNA have been investigated. meso Substituted porphyrins showed the stronger effect on DNA than that of beta substituted porphyrin. Cytotoxicity of compound 3 (IC(50)) to THP-1 tumor cell was up to 0.11 nM. PMID- 15109656 TI - Synthesis of 1alpha,25-dihydroxyvitamin D3-26,23-lactams (DLAMs), a novel series of 1 alpha,25-dihydroxyvitamin D3 antagonist. AB - Novel vitamin D(3) analogs having a lactam structure in their side chains, 1 alpha,25-dihydroxyvitamin D(3)-26,23-lactams (DLAMs), were designed based on the principle of regulation of the folding of helix-12 in the vitamin D nuclear receptor (VDR). The new analogs were synthesized via 1,3-dipolar cycloaddition reaction and subsequent reduction of the isoxazolidine as key steps. Among the analogs, (23S,25S)-DLAM-01 (4a) and (23S,25S)-DLAM-1P (5a) bind strongly to VDR. Moreover, these analogs were found to inhibit the differentiation of HL-60 cells induced by 1 alpha,25-dihydroxyvitamin D(3). PMID- 15109657 TI - Alpha,alpha-trehalose derivatives bearing guanidino groups as inhibitors to HIV-1 Tat-TAR RNA interaction in human cells. AB - Replication of HIV-1 requires specific interactions of Tat protein with TAR RNA. Disruption of Tat-TAR RNA interaction could inhibit HIV-1 replication. Here four target compounds were designed and synthesized to bind to TAR RNA for blocking the interaction of Tat-TAR RNA. The core molecule 6,6'-diamino-6,6'-dideoxy alpha,alpha-trehalose was obtained from selective bromination of, alpha,alpha trehalose at C-6,6', followed by acetylation, azide displacement, deacetylation, and reduction. Coupling of the core molecule with the protected amino acid, then deprotection and guanidinylation generated the novel alpha,alpha-trehalose derivatives. Their abilities to inhibit Tat-TAR RNA interaction in human cells were determined by a Tat-dependent HIV-1 LTR-driven CAT assays. PMID- 15109658 TI - S-(2-(acylamino)phenyl) 2,2-dimethylpropanethioates as CETP inhibitors. AB - Studies on the relationship between the structure of the benzene moiety of S-(2 (acylamino)phenyl) 2,2-dimethylpropanethioates and CETP inhibitory activity were performed. Substituents on the benzene moiety influenced CETP inhibitory activity in a type and position dependent manner, and electron-withdrawing groups at the 4 or 5-position increased the activity. The most potent compound showed 50% inhibition of CETP activity in human plasma at a concentration of 2 microM. PMID- 15109659 TI - Unnatural base pairs mediate the site-specific incorporation of an unnatural hydrophobic component into RNA transcripts. AB - Site-specific incorporation of a hydrophobic nucleotide analog into RNA, by T7 transcription mediated by unnatural base pairs, was developed. The nucleotide analog, 5-phenylethynyl-3-(beta-D-ribofuranosyl)pyridin-2-one 5-triphosphate (denoted by Ph-yTP), was chemically synthesized and then site-specifically incorporated by T7 RNA polymerase into RNA opposite the pairing partner, 2-amino 6-(2-thienyl)purine (denoted by s) in DNA templates. The introduction of Ph-y into a theophylline-binding RNA aptamer, in which a uridine in the internal loop was replaced by Ph-y, raised the thermal stability of the aptamer. Thus, this unnatural nucleotide analog would be useful for stabilizing RNA tertiary structures and complexes between RNA and other molecules. PMID- 15109660 TI - New analogue of arenastatin A, a potent cytotoxic spongean depsipeptide, with anti-tumor activity. AB - Two analogues possessing steric hindered substituents on C-15 of arenastatin A (1), a potent cytotoxic spongean depsipeptide, were synthesized and shown to enhance stability in mouse serum. Notably, 15-tert-butylanalogue (6) with higher cytotoxicity exhibited in vivo anti-tumor activity through iv administration different from 1. Additionally, conformation analysis among the two analogues and arenastatin A (1) indicated that the torsion angle from C-14 to C-20 is a conclusive factor for the potent cytotoxicity of 1. PMID- 15109661 TI - Cycloalkyl[b][1,4]benzodiazepinoindoles are agonists at the human 5-HT2C receptor. AB - Evaluation of selected compounds from our Corporate Compound Library in a human 5 HT(2C) receptor binding assay led to the discovery of WAY-629, a cyclohexyl[b][1,4]benzodiazepinoindole (K(i) 56 nM, E(max) 90%), which is selective for the 5-HT(2C) receptor versus other serotonin receptor subtypes, and dopamine, histamine, adrenergic, and muscarinic receptors. In addition, WAY-629 was active in vivo in a rat model of feeding behavior. An SAR study based on WAY 629 led to compound 11 (K(i) 13 nM, E(max) 102%). PMID- 15109662 TI - Unexpected role of 5-OH in DPPH radical-scavenging activity of 4-thiaflavans. Revealed by theoretical calculations. AB - The O-H bond dissociation enthalpies (BDEs) of 4-thiaflavans were calculated by a combined density functional theory method (RO)B3LYP/6-311 + G(2d,2p)//AM1/AM1. The calculated BDEs not only gave a reasonable explanation on the DPPH radical scavenging activity difference of 4-thiaflavans, but also revealed the unexpected role of 5-OH in enhancing the antioxidant activity of A-ring. PMID- 15109663 TI - A-420983: a potent, orally active inhibitor of lck with efficacy in a model of transplant rejection. AB - We have identified the pyrazolo[3,4-d]pyrimidine A-420983 (compound 7) as a potent inhibitor of lck. A-420983 exhibits oral efficacy in animal models of delayed-type hypersensitivity and organ transplant rejection. PMID- 15109664 TI - Three new cyclostellettamines, which inhibit histone deacetylase, from a marine sponge of the genus Xestospongia. AB - Three new cyclostellettamines, cyclostellettamine G (1), dehydrocyclostellettamines D (2), and E (3), were isolated together with the known cyclostellettamine A (4) from a marine sponge of the genus Xestospongia as histone deacetylase inhibitors. Their structures were determined by spectral and chemical methods. They inhibit histone deacetylase derived from K562 human leukemia cells with IC(50) values ranging from 17 to 80 microM. PMID- 15109665 TI - Efficient synthesis and structure-activity relationship of honokiol, a neurotrophic biphenyl-type neolignan. AB - Honokiol, a biphenyl-type neolignan, which shows the remarkable neurotrophic effect in primary cultured rat cortical neurons, has been effectively synthesized in 21% yield over 14 steps starting from 5-bromosalicylic acid and p hydroxybenzoic acid by utilizing Pd-catalyzed Suzuki-Miyaura coupling reaction as a key step. Additionally, the structure-activity relationship between neurite outgrowth-promoting activity and its O-methylated and/or its hydrogenated analogues was examined in the primary cultures of fetal rat cortical neurons, suggesting that 5-allyl and 4'-hydroxyl groups are essential for affecting the neurotrophic activity of honokiol. PMID- 15109666 TI - Studies on anti-Helicobacter pylori agents. Part 3: A novel, efficacious cephem derivative, FR193879. AB - The synthesis, therapeutic efficacy against H. pylori, and preliminary safety of the novel cephem derivative, FR193879 (8a) are described. Compound 8a having a (4 carbamoylmethylthiazol-2-yl)thio moiety at the 3-position and a phenylacetamido at the 7-position was found to have good safety showing a nontoxic dose of > 100 mg/kg in dogs in a 4-week repeat dose toxicity study and extremely potent therapeutic efficacy against H. pylori, showing 30 times superior activity compared to AMPC, and did not display cross-resistance with CAM or MNZ. PMID- 15109667 TI - Propargylic sulfones possessing a 2-nitroimidazole function: novel hypoxic-cell radiosensitizers with intracellular non-protein thiol depletion ability. AB - Propargylic sulfones (1a-c) containing a 2-nitroimidazole structure were synthesized, and their non-protein thiol (NPSH) depletion abilities were investigated. Propargylic sulfones 1a,c containing an electron withdrawing p nitrophenyl group showed high reactivity toward capturing glutathione (GSH), a typical intracellular NPSH, in phosphate buffer. Among the three propargylic sulfones 1a-c, carboxylic acid derivative 1c showed the most potent radiosensitizing activity toward hypoxic EMT6/KU tumor cells. In view of these results and the partition coefficients between 1-octanol and water, we concluded that appropriate NPSH-depletion ability and lipophilicity are both important in achieving potent hypoxic-cell radiosensitization by propargylic sulfones possessing a 2-nitroimidazole function in biological systems. PMID- 15109668 TI - New cationic lipids for gene transfer with high efficiency and low toxicity: T shape cholesterol ester derivatives. AB - New degradable cationic ester lipids with 'T-shape' configurations were synthesized and tested for gene delivery carrier. Their transfection efficiency and toxicity were compared with commercially available cationic lipids, DOTMA, DOSPA, and DC-Chol. They showed efficient transfection activity and almost no toxicity on mammalian cell lines. Their ester bond degradation was monitored by (1)H NMR. PMID- 15109669 TI - Novel lopinavir analogues incorporating heterocyclic replacements of six-member cyclic urea--synthesis and structure-activity relationships. AB - The HIV protease inhibitor ABT-378 (lopinavir) has a six-member cyclic urea in the P-2 position. A series of analogues in which the six-member cyclic urea is replaced by various heterocycles was synthesized and the structure-activity relationships explored. PMID- 15109670 TI - Synthesis and preliminary biological evaluation of truncated zoanthenol analogues. AB - Zoanthamines are a family of marine alkaloids that have complex heptacyclic structures and are reported to be interleukin-6 modulators. While the structure of zoanthamines, especially the ABC-ring portion, is similar to that of steroids, the CDEFG-ring portion, composed of aminoacetal and lactone core, is a unique structural element. In this report, we designed and synthesized ABC-ring 6 and CDEFG-ring 7, which are truncated analogues of the northern and southern hemispheres of zoanthenol 5, respectively, and which incorporate all of the functionality of each hemisphere. A preliminary SAR study suggested that the hydrochloride of the CEFG-ring portion is an active pharmacophore for suppressing the growth of interleukin-6-dependent MH60 cells. PMID- 15109671 TI - Advances toward new antidepressants beyond SSRIs: 1-aryloxy-3-piperidinylpropan-2 ols with dual 5-HT1A receptor antagonism/SSRI activities. Part 4. AB - A series of 1-(1H-indol-4-yloxy)-3-(4-arylpiperidinyl)propan-2-ols possessing potent dual 5-HT(1A) receptor antagonism and serotonin reuptake inhibition was discovered. The fused aryl ring moiety contributed to the robust dual activities irrespective of the regiochemistry associated with its connectivity to the piperidine central ring. PMID- 15109672 TI - Synthesis of fluorescein labeled 7-methylguanosinemonophosphate. AB - Binding of eIF4E to the cap structure (m(7)GpppN) plays a critical role in mRNA translation. To study the interaction between eIF4E and cap, and to identify small molecule inhibitors of their binding, we synthesized a fluorescent-labeled cap analogue and used it to develop a fluorescence-polarization assay. This preliminary communication describes the synthesis of a fluorescein labeled 7 methylguanosinemonophosphate, and its dose dependent binding to purified human eIF4E as demonstrated by the fluorescence polarization assay. PMID- 15109673 TI - Lead discovery of quinoxalinediones as an inhibitor of dipeptidyl peptidase-IV (DPP-IV) by high-throughput screening. AB - N-Ureido-quinoxalinedione derivatives have been discovered as leads for a novel series of dipeptidyl peptidase-IV (DPP-IV) inhibitors through high-throughput screening of our chemical library. A brief structure-activity relationship of the compounds was investigated. Among them, entry 5 showed the most potent inhibitory activity. The nitro group in quinoxaline moiety and the aromatic sulfonyl substituted ureido functional group seem to be important to increase the potency dramatically. PMID- 15109674 TI - Highly-efficient DNA photocleavers with long wavelength absorptions: thio heterocyclic fused naphthalimides containing aminoalkyl side chains. AB - Thio-heterocyclic fused naphthalimides with aminoalkyl side chains were designed, synthesized and evaluated. These compounds have long wavelength absorptions and binding affinities to Calf thymus DNA. They could photodamage supercoiled pBR322 DNA from form I (closed) to II (nicked) at a concentration as low as 0.5 microM and to form III (linear) at a concentration of 50 microM. A possible mechanism of superoxide anion was provided. PMID- 15109675 TI - The effect of C2-fluoro group on the biological activity of DC-81 and its dimers. AB - C2-Fluoro substituted pyrrolobenzodiazepines (PBDs) have been synthesized that exhibit potential anticancer activity in a number of human tumour cell lines. These C2-fluoro substituted PBDs also exhibit significant DNA-binding ability. PMID- 15109676 TI - Design, synthesis and evaluation of potential inhibitors of HIV gp120-CD4 interactions. AB - This paper describes an approach to prevent HIV-cell fusion by disrupting the interaction between HIV protein gp120 and CD4 receptor. The CD4 residues Phe43 and Arg59 make important interactions with gp120. Small molecule analogues were made to mimic the crucial features of these residues. The analogues were assayed using a cellular 'FIGS' assay to measure inhibition of cell fusion and caused some inhibition. PMID- 15109677 TI - Threshold interaction energy of NRTI's (2'-deoxy 3'-substituted nucleosidic analogs of reverse transcriptase inhibitors) to undergo competitive inhibition. AB - A quantum pharmacological study has been carried out on nucleosidic inhibitors for HIV-1RT where ab initio HF molecular orbital calculations in conjunction with other quantum mechanical techniques have been utilized in a systematic manner to understand the pharmacophoric features and evaluate specific drug-receptor interactions. The interaction energy between the drug and the closest asp 185 of the catalytic triad has been indicated to be crucial in determining the potency of the nucleosidic drug. This study also emphasizes on identifying important specific drug-receptor interactions and evaluating them at the microscopic level to understand the potency regulation as minor conformational changes may lead to significant difference in interaction energies. Although based on relatively few points our correlation of interaction energies with potency data indicates requirement of approximately 13 kcal/mol threshold interaction energy for the drug to undergo efficient competitive inhibition. PMID- 15109678 TI - A new synthesis of indole 5-carboxylic acids and 6-hydroxy-indole-5-carboxylic acids in the preparation of an o-hydroxylated metabolite of vilazodone. AB - A major metabolite of the potential antidepressant vilazodone formed in rat, dog, monkey and human liver microsomes is the 5-cyano-6-hydroxy-1H-indole derivative. For the construction of the salicyl-like substituted indole we adapted a synthesis of carmoxirole using Japp-Klingemann type Fischer-indole synthesis protocols. Functional group interconversion of carboxylic acid via carboxamide into cyanide was performed with methanesulfonic acid chloride. PMID- 15109679 TI - Oxamyl dipeptide caspase inhibitors developed for the treatment of stroke. AB - Structural modifications were made to a previously described acyl dipeptide caspase inhibitor, leading to the oxamyl dipeptide series. Subsequent SAR studies directed toward the warhead, P2, and P4 regions of this novel peptidomimetic are described herein. PMID- 15109680 TI - How laminar frontal cortex and basal ganglia circuits interact to control planned and reactive saccades. AB - How does the brain learn to balance between reactive and planned behaviors? The basal ganglia (BG) and frontal cortex together allow animals to learn planned behaviors that acquire rewards when prepotent reactive behaviors are insufficient. This paper proposes a new model, called TELOS, to explain how laminar circuitry of the frontal cortex, exemplified by the frontal eye fields, interacts with the BG, thalamus, superior colliculus, and inferotemporal and parietal cortices to learn and perform reactive and planned eye movements. The model is formulated as fourteen computational hypotheses. These specify how strategy priming and action planning (in cortical layers III, Va and VI) are dissociated from movement execution (in layer Vb), how the BG help to choose among and gate competing plans, and how a visual stimulus may serve either as a movement target or as a discriminative cue to move elsewhere. The direct, indirect and hyperdirect pathways through the BG are shown to enable complex gating functions, including deferred execution of selected plans, and switching among alternative sensory-motor mappings. Notably, the model can learn and gate the use of a What-to-Where transformation that enables spatially invariant object representations to selectively excite spatially coded movement plans. Model simulations show how dopaminergic reward and non-reward signals guide monkeys to learn and perform saccadic eye movements in the fixation, single saccade, overlap, gap, and delay (memory-guided) saccade tasks. Model cell activation dynamics quantitatively simulate seventeen established types of dynamics exhibited by corresponding real cells during performance of these tasks. PMID- 15109681 TI - ARTSTREAM: a neural network model of auditory scene analysis and source segregation. AB - Multiple sound sources often contain harmonics that overlap and may be degraded by environmental noise. The auditory system is capable of teasing apart these sources into distinct mental objects, or streams. Such an 'auditory scene analysis' enables the brain to solve the cocktail party problem. A neural network model of auditory scene analysis, called the ARTSTREAM model, is presented to propose how the brain accomplishes this feat. The model clarifies how the frequency components that correspond to a given acoustic source may be coherently grouped together into a distinct stream based on pitch and spatial location cues. The model also clarifies how multiple streams may be distinguished and separated by the brain. Streams are formed as spectral-pitch resonances that emerge through feedback interactions between frequency-specific spectral representations of a sound source and its pitch. First, the model transforms a sound into a spatial pattern of frequency-specific activation across a spectral stream layer. The sound has multiple parallel representations at this layer. A sound's spectral representation activates a bottom-up filter that is sensitive to the harmonics of the sound's pitch. This filter activates a pitch category which, in turn, activates a top-down expectation that is also sensitive to the harmonics of the pitch. Resonance develops when the spectral and pitch representations mutually reinforce one another. Resonance provides the coherence that allows one voice or instrument to be tracked through a noisy multiple source environment. Spectral components are suppressed if they do not match harmonics of the top-down expectation that is read-out by the selected pitch, thereby allowing another stream to capture these components, as in the 'old-plus-new heuristic' of Bregman. Multiple simultaneously occurring spectral-pitch resonances can hereby emerge. These resonance and matching mechanisms are specialized versions of Adaptive Resonance Theory, or ART, which clarifies how pitch representations can self-organize during learning of harmonic bottom-up filters and top-down expectations. The model also clarifies how spatial location cues can help to disambiguate two sources with similar spectral cues. Data are simulated from psychophysical grouping experiments, such as how a tone sweeping upwards in frequency creates a bounce percept by grouping with a downward sweeping tone due to proximity in frequency, even if noise replaces the tones at their intersection point. Illusory auditory percepts are also simulated, such as the auditory continuity illusion of a tone continuing through a noise burst even if the tone is not present during the noise, and the scale illusion of Deutsch whereby downward and upward scales presented alternately to the two ears are regrouped based on frequency proximity, leading to a bounce percept. Since related sorts of resonances have been used to quantitatively simulate psychophysical data about speech perception, the model strengthens the hypothesis that ART-like mechanisms are used at multiple levels of the auditory system. Proposals for developing the model to explain more complex streaming data are also provided. PMID- 15109682 TI - Some sufficient conditions for global exponential stability of delayed Hopfield neural networks. AB - In this paper, we have derived some sufficient conditions for existence and uniqueness of equilibrium and global exponential stability in delayed Hopfield neural networks by using a different approach from the usually used one where the existence, uniqueness of equilibrium and stability are proved in two separate steps, rather we first prove global exponential convergence to 0 of the difference between any two solutions of the original neural networks, the existence and uniqueness of equilibrium is the direct results of this procedure. We obtain the conditions by suitable construction of Lyapunov functionals and estimation of derivates of the Lyapunov functionals by the well-known Young's inequality and Holder's inequality. The proposed conditions are related to p norms of vector or matrix, p in [1, infinity] and thus unify and generalize some results in the literature. PMID- 15109683 TI - Bifurcation analysis on a two-neuron system with distributed delays in the frequency domain. AB - In this paper, a general two-neuron model with distributed delays and a strong kernel is investigated. By applying the frequency domain approach and analyzing the associated characteristic equation, the existence of bifurcation parameter for the model is determined. Furthermore, if the mean delay used as a bifurcation parameter, it is found that Hopf bifurcation occurs for the strong kernel. This means that a family of periodic solutions bifurcates from the equilibrium when the bifurcation parameter exceeds a critical value. The direction and stability of the bifurcating periodic solutions are determined by the Nyquist criterion and the graphical Hopf bifurcation theorem. Some numerical simulations are given to justify the theoretical analysis results. PMID- 15109684 TI - The MDF discrimination measure: Fisher in disguise. AB - Recently, a discrimination measure for feature extraction for two-class data, called the maximum discriminating (MDF) measure (Talukder and Casasent [Neural Networks 14 (2001) 1201-1218]), was introduced. In the present paper, it is shown that the MDF discrimination measure produces exactly the same results as the classical Fisher criterion, on the condition that the two prior probabilities are chosen to be equal. The effect of unequal priors on the efficiency of the measures is also discussed. PMID- 15109685 TI - GenSo-EWS: a novel neural-fuzzy based early warning system for predicting bank failures. AB - Bank failure prediction is an important issue for the regulators of the banking industries. The collapse and failure of a bank could trigger an adverse financial repercussion and generate negative impacts such as a massive bail out cost for the failing bank and loss of confidence from the investors and depositors. Very often, bank failures are due to financial distress. Hence, it is desirable to have an early warning system (EWS) that identifies potential bank failure or high risk banks through the traits of financial distress. Various traditional statistical models have been employed to study bank failures [J Finance 1 (1975) 21; J Banking Finance 1 (1977) 249; J Banking Finance 10 (1986) 511; J Banking Finance 19 (1995) 1073]. However, these models do not have the capability to identify the characteristics of financial distress and thus function as black boxes. This paper proposes the use of a new neural fuzzy system [Foundations of neuro-fuzzy systems, 1997], namely the Generic Self-organising Fuzzy Neural Network (GenSoFNN) [IEEE Trans Neural Networks 13 (2002c) 1075] based on the compositional rule of inference (CRI) [Commun ACM 37 (1975) 77], as an alternative to predict banking failure. The CRI based GenSoFNN neural fuzzy network, henceforth denoted as GenSoFNN-CRI(S), functions as an EWS and is able to identify the inherent traits of financial distress based on financial covariates (features) derived from publicly available financial statements. The interaction between the selected features is captured in the form of highly intuitive IF-THEN fuzzy rules. Such easily comprehensible rules provide insights into the possible characteristics of financial distress and form the knowledge base for a highly desired EWS that aids bank regulation. The performance of the GenSoFNN-CRI(S) network is subsequently benchmarked against that of the Cox's proportional hazards model [J Banking Finance 10 (1986) 511; J Banking Finance 19 (1995) 1073], the multi-layered perceptron (MLP) and the modified cerebellar model articulation controller (MCMAC) [IEEE Trans Syst Man Cybern: Part B 30 (2000) 491] in predicting bank failures based on a population of 3635 US banks observed over a 21 years period. Three sets of experiments are performed-bank failure classification based on the last available financial record and prediction using financial records one and two years prior to the last available financial statements. The performance of the GenSoFNN-CRI(S) network as a bank failure classification and EWS is encouraging. PMID- 15109686 TI - New training strategies for constructive neural networks with application to regression problems. AB - Regression problem is an important application area for neural networks (NNs). Among a large number of existing NN architectures, the feedforward NN (FNN) paradigm is one of the most widely used structures. Although one-hidden-layer feedforward neural networks (OHL-FNNs) have simple structures, they possess interesting representational and learning capabilities. In this paper, we are interested particularly in incremental constructive training of OHL-FNNs. In the proposed incremental constructive training schemes for an OHL-FNN, input-side training and output-side training may be separated in order to reduce the training time. A new technique is proposed to scale the error signal during the constructive learning process to improve the input-side training efficiency and to obtain better generalization performance. Two pruning methods for removing the input-side redundant connections have also been applied. Numerical simulations demonstrate the potential and advantages of the proposed strategies when compared to other existing techniques in the literature. PMID- 15109687 TI - Introduction: guidelines and advisory development. AB - Since 1992, the American Society of Anesthesiologists has produced 12 evidence based practice guidelines, 2 practice advisories, and 3 guideline updates. These documents have assisted anesthesiologists and practitioners in many other specialties. Their brevity, practicality, and ease of use, coupled with a thorough and systematic evaluation of the evidence have been instrumental in bringing together the science and practice of medicine. The application of formal evidence-collection processes for literature and opinion and efficient analytic evaluations combine with the experience and practical knowledge of clinicians to produce widespread application of the guidelines. The evidence-based process developed by the ASA has been found to be adaptable to a wide variety of issues relating to clinical practice. The goal is to systematically collect and evaluate evidence from multiple sources and apply it ina comprehensive manner to the guideline recommendations. The ASA guideline and advisory development process is continuing to evolve in response to changes in medical technology, research, and practice. By providing synthesized evidence from multiple sources and robust clinical recommendations the ASA offers the practice of anesthesiology, an invaluable bridge between science and clinical practice. PMID- 15109688 TI - Preoperative laboratory testing: general issues and considerations. AB - The challenge of preoperative laboratory testing has been increased during the past 20 years because now fewer than 30% of patients undergoing surgery are admitted to the hospital before the day of surgery. Even with the advent of formal preoperative systems in many institutions, it is believed that well over 50% of patients do not have a definitive preoperative visit with anesthesia staff before the day of surgery. Thus, anesthesia teams are dependent on other providers to provide appropriate data to best judge who is an appropriate candidate for anesthesia. This issue is becoming all the more difficult because the acuity of patients and complexity of procedures being managed on an outpatient or same day admission basis increases. PMID- 15109689 TI - Age-associated issues in preoperative evaluation, testing, and planning: pediatrics. AB - The author has reviewed recent developments in preoperative assessment and testing, emphasizing issues that are of greatest concern in pediatric patients. Attention to these areas during the process of preoperative preparation and appropriate communication of conditions that may contribute to increased peri operative risk will provide greater predictability for families, surgeons, and operating room staff. This predictability is an important component to improved patient or family satisfaction and operating room efficiency. PMID- 15109690 TI - Age associated issues: geriatrics. AB - The effects of aging on the body are numerous, but the most important point with regard to surgery is to differentiate the effects of aging from those of the pathology associated with a disease process that may also be present in the elderly patient. Studies tend to validate the premise that there are no labs mandated solely because of a patient's age. Rather, the physician should analyze the geriatric patient with reference to the nature of the surgery emergent versus elective; the degree of risk for the surgical procedure itself-that is high-risk, intermediate--risk, or low-risk surgery; while bearing in mind the patient's overall physiologic state. In assessing a patient's overall physiologic state an organ systems based approach focusing on the cardiac, respiratory, renal, hepatic, endocrine, nutritional, and neurologic systems may be warranted and beneficial. In the elderly population one of the key predictors of perioperative complications seems to be the geriatric patient's preoperative condition and preoperative level of functioning. PMID- 15109691 TI - Preoperative cardiac evaluation. AB - Guidelines on perioperative cardiovascular evaluation for noncardiac surgery have been published. The integration of clinical risk factors, surgery-specific risk,and functional capacity should be used to determine the need for further diagnostic evaluation. The use of beta-adrenergic blockade in high-risk patients,particularly those with documented myocardium at risk undergoing vascular surgery, has been shown to reduce perioperative risk and may obviate the need for more invasive procedures. Coronary intervention should be reserved for those patients who warrant intervention independent of the noncardiac surgery. PMID- 15109692 TI - Preoperative assessment: pulmonary. AB - Understanding the risk factors for the development of PPCs allows targeted interventions aimed at reducing the frequency and severity of PPCs. The broad categories of what increases the likelihood of developing a PPC are understood but specific understanding of how individual risk factors act to cause PPCs is lacking,and there is little information regarding the interaction or synergy between risk factors. Further research is needed to define the nature of risk factors and develop better predictive models of patients at risk for developing PPCs. It is clear that anesthetic agents produce significant changes in the respiratory system but further information is needed to define how such changes contribute, if at all, to the subsequent development of PPCs. The ongoing controversy regarding the value of regional analgesia or anesthetic techniques, especially epidural analgesia and anesthesia, in reducing or preventing PPCs requires well-done randomized clinical trials. Further research is also needed in the area of postoperative care such as interventions in patients with OSA or the use of inventive spirometric techniques. PMID- 15109693 TI - Assessment and therapy of selected endocrine disorders. AB - Diabetes remains the most commonly encountered endocrinopathy with the incidence of type 2 doubling in the past decade. The prevalence of diabetes is projected to continue to increase dramatically over the next several decades unless major public health initiatives are successful in stemming this growth. Both type I and 2 diabetics more frequently require surgical and critical care than their non diabetic counterparts. Type 1 and 2 diabetics also sustain greater peri-operative morbidity and mortality. Careful preoperative assessment and appropriate perioperative intervention may limit this. There is increasing evidence that maintenance of normal blood glucose in the perioperative period and during critical illness is beneficial for diabetic and non-diabetic patients. More data will hopefully be forthcoming to substantiate recent reports and identify the mechanisms of improved outcome. Thyroid disease remains a commonly encountered pathology that is more readily identified and controlled in the modern era of radioimmune assays of thyroid hormone and successful medical and surgical therapies. Severe hypothyroidism and thyroid storm are associated with significant increases in perioperative morbidity and mortality. Recognition of these entities or those at risk for developing them post operatively is crucial in initiating timely and effective therapy. Primary Al is uncommon, but results in glucocorticoid and mineralocorticoid deficiency. Tertiary Al is far more common, most often secondary to iatrogenic therapy with exogenous glucocorticoids for the management of chronic diseases such as connective tissue disorders, anti rejection regimes, and severe asthma. Glucocorticoid replacement or supplementation is needed on a case-by-case basis and should be individualized based on chronic steroid dose, duration, and stress of the surgical procedure. Perioperative steroid dosing regimes now recommend lower doses for shorter periods than previously suggested. More recently Al has been recognized in two populations, elderly patients undergoing major surgery and a subgroup of patients with septic shock. Timely diagnosis using synthetic ACTH stimulation testing and stress glucocorticoid, and possibly mineralocorticoid therapy, seems to reverse these processes and improve recovery. Although uncommon, patients with pheochromocytoma who undergo open or laparoscopic resections remain diagnostic and therapeutic challenges. Perioperative outcome seems to have improved, in part, related to newer therapies and less invasive surgeries when indicated. The appropriate preoperative assessment and management of patients with various endocrinopathies is important to optimize outcome and limit avoidable complications. Hopefully additional evidence based guidelines will be forth coming particularly in caring for the ever increasingly encountered perioperative diabetic. PMID- 15109694 TI - Perioperative anesthesia clinical considerations of alternative medicines. AB - The presence of nutraceutical agents in the United States health care system dictates the need for a general understanding of these agents by all physicians and health care providers. Increasing trend toward reimbursement of herbal medicines by the insurance companies and managed care organizations have further encouraged their use. Because herbs are listed under the "supplement" category by the Food and Drug Administration, the Dietary Supplement and Health Education Act establishes no protocol for standardization of the products labeled as "supplements" thereby increasing the risk for adverse effects associated with the use of these products. Moreover, there is little motivation for the manufacturers to conduct randomized, placebo-controlled, double-blinded safety and efficacy trials on these drugs. Reports indicate that within the last 2 decades,more than 100 herbogenic deaths have occurred [100]. Many serious complications have been reported, including renal failure and need for renal or hepatic transplantation after taking nutraceuticals [101-107]. The anesthesiologists should have a detailed knowledge and understanding of the potential risks and purported benefits of herbal medicines and should thoroughly inquire about patient's use of herbal products [108- 11 ]. In addition,the education of each patient regarding the serious, potential drug-herb inter-actions should be a routine component of preoperative assessment. The American Society of Anesthesiologists (ASA), suggests that all herbal medications should be discontinued 2 to 3 weeks before an elective surgical procedure. If the patient is not sure of the contents of the herbal medicine, he or she should be urged to bring the container so that the anesthesiologist can review the contents of the herb or preparation. Although this idea holds some promise in the elective-care settings, anesthetic care in emergency settings should be based on a thorough drug-intake history from the patient or a relative, if possible.Unfortunately, the anesthesia literature has not addressed this new group of health supplements, despite many of these drugs having the potential to cause serious health problems and drug-herb interactions. There is a need to conduct additional scientific clinical trials to study the anesthetic responses to commonly used nutraceutical agents. PMID- 15109695 TI - Preanesthetic evaluation in private practice. AB - The preanesthetic evaluation is an essential element of the anesthetic management of a patient. The type of practice, academic or private, should have no bearing on the quality of the evaluation. The practice environment may play a significant role in the type of patient and procedure but should not diminish the quality of care for any particular patient. However the extent, timing methodologies for the preanesthetic evaluation will vary according to patient medical requirements, surgical procedures, resource availability, patient convenience and,perhaps the key element, staffing availability. Each practice environment needs to develop its own solutions. Private practice, especially in the smaller locations,may need to be more sensitive to staffing and personnel issues and patient satisfaction. Technology, especially in the smaller private practice environments where staffing is more likely to be an issue, may greatly facilitate the process and improve the quality. However, technology may require extensive support (time and money) to implement, but the indirect gains should justify the effort. PMID- 15109696 TI - Quality and resource utilization in managing preoperative evaluation. AB - The evaluation of patients before surgery is a component of anesthesia practice that must be performed to ensure the safe delivery of anesthesia for every patient. How an anesthesia group performs this evaluation differs from institution to institution as the structure of preoperative clinics in hospitals across the country can be quite variable. However, the major question that anesthesiologists must ask when approaching this area of practice is: "Can a system for preoperative evaluation that uses the expertise of anesthesiologists,surgeons, and internists to deliver high quality, cost conscious, and efficient patient preparation for surgery be developed. PMID- 15109697 TI - Preoperative evaluation. PMID- 15109698 TI - Preoperative evaluation, testing, and planning. PMID- 15109699 TI - A population study of the precedence effect. AB - Data are reported from a population of untrained individuals under lag- and single-click conditions in a discrimination suppression precedence-effect task. The cue to be discriminated was an interaural level-difference (ILD). Each of 91 observers completed 10 runs in a two-interval forced-choice design under a lag click condition and three runs under a single-click condition. Stimuli were 125 micros rectangular pulses and the interclick interval was 2 ms. Observers were randomly assigned to three groups of approximately 30. Each group was then tested at one stimulus intensity (43, 58, or 73 dB). Mean threshold within each group was greater than 15 dB for the lag-click condition and 6 dB for the single-click condition, although there was substantial interobserver variability. In contrast to [J. Acoust. Soc. Am. 114 (2003) 420] who reported a strong effect of intensity on lag-click ITD discrimination, no effect of intensity was observed on lag-click ILD thresholds. Analysis of over 50,000 near-threshold trials from 302 observers pooled across studies showed a spatial asymmetry in response patterns and a small, but statistically significant effect of gender. A model is proposed which shows that decay of sensory memory and increases in auditory filter bandwidths with intensity may predict the different findings for ILD versus ITD lag-click thresholds. PMID- 15109700 TI - Spatial discrimination of sound sources in the horizontal plane following an adapter sound. AB - The effect of a preceding (adapter) sound on the spatial discrimination of two subsequent, successively presented (target) sounds was tested in the horizontal plane. The adapter and the first target were located in front of the subject or 30 degrees to the right of the midline; both sounds were presented either at the same location or at different locations. The second target was located to the right or the left of the first. Sound spectra of the 3-s adapter and the 100-ms targets were either high (4.5-18 kHz) or low (1-4 kHz) in frequency. Fifteen subjects judged the position of the second target relative to the first in a two alternative forced-choice paradigm. In comparison with a no-adapter control condition, in which no sound preceded, discrimination performance was increased when adapter and first target were presented at the same location and when both sounds consisted of the same frequency spectrum. No improvement occurred when adapter and targets differed in location or frequency. The results are consistent with previous results on post-adaptation discrimination of interaural time differences. Possibly, spatial adaptation of the underlying mechanisms of auditory localization may explain the discrimination aftereffect. PMID- 15109701 TI - The effects of aging in the medial geniculate nucleus: a comparison with the inferior colliculus and auditory cortex. AB - A common problem among the elderly is a difficulty in discriminating speech. One factor that may contribute to this is deterioration in the ability to process the dynamic components of speech such as formant transitions. The frequency-modulated (FM) sweep is a useful stimulus for investigating the neural basis of temporal processing speed since it has features in common with formant transitions. Previously, we showed that when cells in the auditory cortex of aged animals were presented with FM sweeps, they exhibited a decrease in temporal processing speed when compared to cells recorded from young animals. However, this was not the case for cells in the inferior colliculus (IC) where neural responses did not appear to be affected by aging. One question that remains is how the auditory thalamus is affected by aging: Is it similar to that of the auditory cortex or of the IC. To this end, single units were recorded from the ventral division of the medial geniculate nucleus (MGNv) of young and aged anaesthetized rats in response to FM sweeps. Results showed that there were no age-related differences in speed or direction selectivity of FM sweep responses in the MGNv. When compared with units recorded from the IC and AI, the responses of MGNv neurons were similar to those of the IC. This suggests that temporal processing speed is affected by aging in the cortex, but not in the auditory thalamus or midbrain. PMID- 15109702 TI - Gravity receptor function in mice with graded otoconial deficiencies. AB - The purpose of the present study was to examine gravity receptor function in mutant mouse strains with variable deficits in otoconia: lethal milk (lm), pallid (pa), tilted (tlt), mocha (mh), and muted (mu). Control animals were either age matched heterozygotes or C57BL/6J (abbr. B6) mice. Gravity receptor function was measured using linear vestibular evoked potentials (VsEPs). Cage and swimming behaviors were also documented. Temporal bones were cleared to assess the overall otoconial deficit and to correlate structure and function for lm mice. Results confirmed the absence of VsEPs for mice that lacked otoconia completely. VsEP thresholds and amplitudes varied in mouse strains with variable loss of otoconia. Some heterozygotes also showed elevated VsEP thresholds in comparison to B6 mice. In lm mice, which have absent otoconia in the utricle and a variable loss of otoconia in the saccule, VsEPs were present and average P1/N1 amplitudes were highly correlated with the average loss of saccular otoconia (R = 0.77,p < 0.001). Cage and swimming behavior were not adversely affected in those animals with recordable VsEPs. Most, but not all, mice with absent VsEPs were unable to swim. Some animals were able to swim despite having no measurable gravity receptor response. The latter finding underscores the remarkable adaptive potential exhibited by neurobehavioral systems following profound sensory loss. It also shows that behavior alone may be an unreliable indicator of the extent of gravity receptor deficits. PMID- 15109703 TI - Platelet GPIaC807T polymorphism is associated with negative outcome of sudden hearing loss. AB - To determine the relevance of inherited prothrombotic risk factors in sudden hearing loss, we investigated 85 patients with sudden hearing loss of >/= 60 dB for the presence of inherited prothrombotic risk factors. The FV G1691A, FII G20210A, GPIa C807T, GPIIIa PIA1/A2, PAI-1 4G/5G, t-PA Alu repeat ID, MTHFR C677T and CBS 844ins68 genotypes were investigated. Allele frequencies found in patients were compared to those of 85 healthy control subjects of the same ethnic background using Chi-squared and odds-ratio analysis. The frequency of the GPIa807T allele was significantly elevated in patients compared to controls. In addition, allele frequency and genotype distribution of GPIa was significantly elevated in the patient group without recovery after 3 months of sudden hearing loss onset. Allele frequencies of all other prothrombotic risk factors investigated here did not differ from those of the control subjects. The single nucleotide polymorphism of GPIa C807T seems to play a role as a prognostic factor in recovery from sudden hearing loss. PMID- 15109704 TI - Tonotopic organization of the human auditory cortex probed with frequency modulated tones. AB - Using neuromagnetic source imaging, we investigated tonotopic representation and direction sensitivity in the auditory cortex of humans (N = 15). For this purpose, source analysis was undertaken at every single sampling point during the presentation of a frequency-modulated tone (FM) sweeping slowly downward or upward across periods of 3 s duration. Stimuli were selected to target response properties of the central part of the primary auditory cortical field, which has been shown to exhibit sensitivity to distinct FM-sound features as compared to the ventral and dorsal part. Linear mixed-effects model statistics confirm tonotopic gradients in medial-lateral and anterior-posterior directions. The high resolution provided by this method revealed that the relationship between frequency and spatial location of the responding neural tissue is nonlinear. The idea that neurons specifically sensitive to the employed sound characteristics (slow, downward modulation) were activated is supported by the fact that the upward sweep of identical duration produced a different pattern of functional organisation. PMID- 15109705 TI - Earlier and later components of tone burst evoked myogenic potentials. AB - The aim of this study was to further investigate the response rates, thresholds, latencies and amplitudes of the earlier and later components of tone burst evoked myogenic potentials (TEMPs) in conditions of binaural and monaural stimulation and recordings in a group of normal subjects in response to a 500-Hz tone burst. Each subject underwent simultaneous binaural acoustic stimulation with bilateral recording (B-TEMP) initially, then monaural acoustic stimulation with ipsilateral recording (M-TEMP) on another day. The results showed 100%, 100%, 54%, and 23% response rates for the earlier components of B-TEMPs, when using 105, 95, 85, and 75 dB acoustic stimulation, respectively. It exhibited a significant decrease in the response rate when the stimulus intensity was attenuated from 95 to 75 dB. In addition, no significant difference in the response rate of the earlier components existed between B-TEMPs and M-TEMPs. For the later components of B TEMPs obtained from 105, 95, 85, and 75 dB acoustic stimulation, the response rates were 85%, 85%, 62%, and 42%, respectively, higher than those of M-TEMPs significantly. Nevertheless, there were no statistical differences in the mean latencies of each peak (p13, n23, n34, and p44) between B-TEMPs and M-TEMPs under similar stimulus intensity. Meanwhile, no significant relationship existed between the stimulus intensity and the latency, regardless of B-TEMPs or M-TEMPs. Comparing the relative amplitudes between B-TEMPs and M-TEMPs, the former displayed significant smaller p13-n23 amplitude, but larger n34-p44 amplitude. However, the mean thresholds for the earlier and later components between B-TEMPs and M-TEMPs did not differ significantly. In conclusion, monaural acoustic stimulation with ipsilateral recording may elicit larger amplitude of the earlier components, whereas binaural acoustic stimulation with bilateral recording evokes higher response rate and larger amplitude of the later components. Although the nerve pathways for both components are different, the thresholds for these potentials do not differ significantly, indicating that both components may, at least in part, share a common origin, but different pathways. PMID- 15109706 TI - Separate projections from the inferior colliculus to the cochlear nucleus and thalamus in guinea pigs. AB - We used multiple-labeling techniques with retrograde fluorescent tracers to determine whether individual cells in the inferior colliculus project to the medial geniculate body (MG) and the cochlear nucleus (CN) in guinea pigs. Four possible projection patterns were examined: (1) to ipsilateral MG and ipsilateral CN; (2) to ipsilateral MG and contralateral CN; (3) to contralateral MG and ipsilateral CN; and, (4) to contralateral MG and contralateral CN. Following injections of different tracers into two or more sites, no inferior collicular cells were double-labeled from the two contralateral targets and only a few cells were double-labeled from each of the other pairs of targets. The double-labeled cells always totaled < 1% of the single-labeled populations. We conclude that collateral projections from the inferior colliculus to the MG and CN are virtually non-existent. Therefore, the ascending and descending projections to these targets arise from different cells. These cells could potentially receive different inputs and send different information to higher or lower centers of the auditory pathway. PMID- 15109708 TI - Responses of the endolymphatic sac to perilymphatic injections and withdrawals: evidence for the presence of a one-way valve. AB - Although the endolymphatic sac (ES) is thought to be a primary site for endolymph volume regulation, we have limited knowledge of how it responds to volume and pressure changes. In a prior publication, we demonstrated changes of K(+), Na(+) and endolymphatic sac potential (ESP) resulting from volume injections into, and withdrawals from, scala media of the cochlea. In the present study, we compared the influence of injections into and withdrawals from scala tympani of the cochlea on the endolymphatic sac. It is assumed that similar pressure changes are induced in endolymph and perilymph of both the cochlear and vestibular compartments of the ear. Pressure changes induced by the perilymphatic injections and withdrawals did not induce similar K(+) changes in the ES. The majority of perilymph withdrawals caused K(+) and ESP reductions in the sac, but few injections caused any measurable changes in the sac. Pressure measurements from the ES demonstrated that transmission of labyrinthine pressures to the lumen was directionally sensitive, with negative pressure transmitted more effectively than positive. In other experiments, application of infrasonic stimulation to the ear canal resulted in K(+) increase in the ES. These physiological measurements suggest that the endolymphatic duct may be closed by sustained positive pressure in the vestibule but open during pressure fluctuations. Study of the anatomy where the endolymphatic duct enters the vestibule suggests that the membranous sinus of the endolymphatic duct could act as a mechanical valve, limiting the flow of endolymph from the saccule to the endolymphatic sac when pressure is applied. This structure could therefore play an important role in endolymph volume regulation. PMID- 15109707 TI - Development of NMDA R1 expression in chicken auditory brainstem. AB - NMDA receptor subunit 1 (NR1) expression in the chicken cochlear nuclei was examined using immunohistochemistry and quantitative Western blots. An antibody raised in mouse against a highly conserved domain of NR1 recognized the same 115 kDa protein band in chicken brain. Quantitative Western blotting of cochlear nucleus protein showed no significant change in NR1 expression from E18 to adult. The nucleus angularis (NA) initiated NR1 expression before E12 that became more prominent after hatching. NR1-ir first appeared in the nucleus magnocellularis (NM) and nucleus laminaris (NL) at E10. From E12 to E19, NM exhibited a gradient in NR1 expression with medial, higher best frequency cell bodies being more immunoreactive than lateral, lower best frequency cell bodies. This gradient disappeared by E20. The distribution of NR1 in NL also changed during development. NR1 label was present in NL cell bodies between E10 and E13. From E14 onwards, NR1-ir characterized both cell bodies and neuropil. After hatching, NR1-ir levels were higher in NL than NM. The superior olive first expressed NR1 at E12. Neuropil staining was more intense than cell bodies until after hatching. In contrast to the functional decrease observed in mammals and chick, NR1-ir expression remained high in the chicken auditory brainstem into adulthood. Both chickens and rodents retain high levels of NR-1. PMID- 15109709 TI - The cochlear F-box protein OCP1 associates with OCP2 and connexin 26. AB - OCP1 and OCP2 are the most abundant proteins in the organ of Corti. Their distributions map identically to the epithelial gap-junction system, which unites the supporting cell population. Sequence data imply that OCP1 and OCP2 are subunits of an SCF E3 ubiquitin ligase. Consistent with that hypothesis, electrophoretic mobility-shift assays and pull-down assays with immobilized OCP1 demonstrate the formation of an OCP1-OCP2 complex. Sedimentation equilibrium data indicate that the complex is heterodimeric. The coincidence of the OCP1-OCP2 distribution and the epithelial gap-junction system suggests that one or more connexin isoforms may be targets of an SCF(OCP1) complex. Significantly, immobilized OCP1 binds (35)S-labeled connexin 26 (Cx26) produced by in vitro transcription-translation. Moreover, Cx26 can be co-immunoprecipitated from extracts of the organ of Corti by immobilized anti-OCP1, implying that OCP1 and Cx26 may associate in vivo. Given that lesions in the Cx26 gene (GJB2) are the most common cause of hereditary deafness, the OCP1-Cx26 interaction has substantial biomedical relevance. PMID- 15109710 TI - Time response of carboplatin-induced hearing loss in rat. AB - Carboplatin is currently being used as an anticancer drug against human cancers. However, high dose of carboplatin chemotherapy resulted in hearing loss in cancer patients. We have shown that carboplatin-induced hearing loss was related to dose dependent oxidative injury to the cochlea in rat model. However, the time response of ototoxic dose of carboplatin on hearing loss and oxidative injury to cochlea has not been explored. The aim of the study was to evaluate the time response of carboplatin-induced hearing loss and oxidative injury to the cochlea of the rat. Male Wistar rats were divided into two groups of 30 animals each and treated as follows: (1) control (normal saline, i.p.) and (2) carboplatin (256 mg/kg, a single i.p. bolus injection). Auditory brain-evoked responses (ABRs) were recorded before and 1-5 days after treatments. The animals (n = 6) from each group were sacrificed on day 1, 2, 3, 4, and 5 and cochleae were isolated and analyzed. Carboplatin significantly elevated the hearing thresholds to clicks and to 2, 4, 8, 16, and 32 kHz tone burst stimuli only 3-5 days post-treatment. Carboplatin significantly increased nitric oxide (NO), malondialdehyde (MDA) levels and manganese superoxide dismutase (Mn-SOD) activity in the cochlea 4-5 and 3-5 days post-treatment, respectively, indicating enhanced influx of free radicals and oxidative injury to the cochlea. Carboplatin significantly depressed the reduced to oxidized glutathione (GSH/GSSG) ratio, antioxidant enzyme activities such as copper/zinc-superoxide dismutase (CuZn-SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) as well as enzyme protein expressions in the cochlea 3-5 days after treatment. The data suggest that carboplatin-induced hearing loss involves oxidative injury to the cochlea of the rat in a time dependent manner. PMID- 15109711 TI - Cochlear nerve demyelination causes prolongation of wave I latency in ABR of the myelin deficient (md) rat. AB - In this study, we examined the auditory brainstem responses (ABRs), distortion product of otoacoustic emissions (DPOAEs) and cochlear morphology of the myelin deficient (md) rat, which completely lacks central myelin but not peripheral myelin. ABRs showed a marked prolongation not only wave II-IV latencies but also wave I latency. Cochlear nerve fibers near the modiolus lost their myelin halfway into the internal auditory canal. DPOAEs also decreased at a lower frequency of the combined tone. Since nerve fibers ending at the apical turn of the cochlea passed through central portion of the cochlear nerve, wave I prolongation of ABRs and decrease of DPOAEs at a lower frequency might originate mainly from the demyelinated CNS part of the cochlear nerve and efferent olivocochlear bundle in the internal auditory canal. PMID- 15109712 TI - Hexachlorobenzene, a dioxin-like compound, disrupts auditory function in rat. AB - Hexachlorobenzene (HCB) is a dioxin-like compound widely distributed in the environment. In this study, we investigated the effects of HCB on the cochlea. Conscious free-moving rats were given HCB per os daily for 4 weeks at doses of 0.16, 4 or 16 mg/kg in olive oil, whereas the control group received olive oil only. The effects of HCB were evaluated at various time intervals, by measuring auditory nerve acoustic thresholds and plasma thyroid hormone concentration by radioimmunoassay. Histological evaluation involved surface preparation and scanning electron microscopy observations of cochlear hair cells. At a dose of 0.16 mg/kg, HCB induced no loss of acoustic sensitivity, whereas at 4 mg/kg, it induced cochlear sensitivity deficits at the mid-frequencies (2-16 kHz) with complete recovery once treatment was stopped. At a dose of 16 mg/kg, permanent threshold shifts were observed at all frequencies tested (from 1 to 32 kHz). Morphological studies showed no cochlear hair cell loss or alteration of stereocilia. HCB treatment reduced circulating thyroxine concentrations. Thyroidectomy had no effect on cochlear sensitivity in control animals. Thus, HCB is a potent oto-toxicant, and its ototoxicity may be independent of its thyroidal effects. PMID- 15109713 TI - Stability of short sequence repeats and their application for the characterization of Erwinia amylovora strains. AB - A motif of eight nucleotides (GAATTACA) reiterated 3 to 15 times within the PstI fragment of the pEa29 plasmid was found in Erwinia amylovora strains representing a valuable typing method for this pathogen. The stability of short sequence DNA repeat (SSR) numbers was investigated to determine the suitability of this marker for strain differentiation. The number of SSR units was found to be stable under laboratory and certain stress conditions. This meets the requirements for a suitable genetic marker that should be stable upon cultivation of strains. Therefore, this SSR marker was used for strain differentiation from SSR-3 to SSR 15 and a large number of E. amylovora strains from Austria was screened for their SSR numbers for epidemiological identification purposes. Traceability was possible if strains had very high or very low SSR numbers. PMID- 15109714 TI - Analysis of orthologous hrcA genes in Escherichia coli and Bacillus subtilis. AB - The hrcA gene codes for a transcriptional repressor protein interacting with the CIRCE operator thereby reducing expression of the groE operon of more than 120 bacterial species. At least in Bacillus subtilis, the activity of the HrcA protein is modulated by the GroE chaperonin system. We amplified the hrcA gene from five different bacterial species and analyzed its activity in Escherichia coli and Bacillus subtilis. While those from Clostridium acetobutylicum and Staphylococcus aureus turned out to be active, those of Helicobacter pylori, Lactococcus lactis and Thermotoga maritima were inactive in E. coli, but that of T. maritima turned out to repress expression of the reporter gene in B. subtilis. All these results strongly suggest to us a specific recognition of HrcA by the GroE chaperonin system. PMID- 15109715 TI - Characterization of an excreted/secreted antigen form of 14-3-3 protein in Toxoplasma gondii tachyzoites. AB - The 14-3-3 protein was shown to be present into the parasitophorous vacuole of Toxoplasma gondii-infected human monocyte cells and in the excreted/secreted antigens (ESA). The ESA 14-3-3 protein migrates electrophoretically as the cytosol and the main membranous 14-3-3 isoforms. The excretion/secretion of 14-3 3 was not sensitive to cycloheximide, a protein synthesis inhibitor, even at a concentration which inhibited the production of 14-3-3 inside the tachyzoites. Recombinant 14-3-3/GST protein was used to test the presence of 14-3-3 antibodies in different human sera. A positive immunoreactivity was observed with sera corresponding to acute toxoplasmosis and a possible involvement of 14-3-3 in host immunity is discussed. PMID- 15109716 TI - Post-translational processing of Neisseria meningitidis gamma-glutamyl aminopeptidase and its association with inner membrane facing to the cytoplasmic space. AB - We previously demonstrated that gamma-glutamyl aminopeptidase (also called gamma glutamyl transpeptidase) (GGT) of Neisseria meningitidis is involved in the bacterial multiplication in cerebrospinal fluid. To further understand the function of meningococcal GGT, the biochemical properties were investigated in this study. The deduced amino acid sequence in N. meningitidis GGT was 37% identical to that of Escherichia coli GGT and that of Helicobacter pylori GGT, respectively, while a typical signal sequence was not found at the N-terminus of meningococcal GGT. Western blotting using rabbit antiserum against recombinant meningococcal GGT protein demonstrated that the meningococcal GGT is processed into two subunits in N. meningitidis at the conserved amino acid, threonine 427. The experiments on subcellular fractionation suggested that the majority of meningococcal GGT is associated with inner membrane facing to the cytoplasmic side. This cell localization might be unique for N. meningitidis compared to other GGTs. PMID- 15109717 TI - Analysis of spontaneous base substitutions generated in mutator strains of Bacillus subtilis. AB - In the current studies, we investigated base substitutions in the Bacillus subtilis mutT, mutM, and mutY DNA error-prevention system. In the wild type strain, spontaneous mutations were mainly transitions, either G:C --> A:T or A:T -> G:C. Although both transitions and transversions were observed in mutY and mutM mutants, mutM/mutY double mutants contain strictly G:C --> T:A transversions. In the mutT strain, A:T --> C:G transversion was not observed, and over-expression of the B. subtilis mutT gene had no effect on the mutation rate in the Escherichia coli mutT strain. Using 8-oxo-dGTP-induced mutagenesis, transitions especially A:T --> G:C were predominant in the wild type and mutY strains. In contrary, transversion was high on mutY and double mutant (mutM mutY). Finally, the opuBC and yitG genes were identified from the B. subtilis chromosome as mutator genes that prevented the transition base substitutions. PMID- 15109718 TI - Molecular characterization and analysis of the operon encoding the antifungal lipopeptide bacillomycin D. AB - Bacillus subtilis AU195 produces bacillomycin D, a cyclic lipopeptide that is an inhibitor of the aflatoxin producing fungus Aspergillus flavus. Sequence analysis of the bacillomycin D operon revealed four ORFs with the structural organization of the peptide synthetases. Disruption of ORF 2, which links the amino acid moiety to the b-amino fatty acid, resulted in the loss of antifungal activity. By comparing the sequence of bacillomycin D, iturin A and mycosubtilin operons, our results showed that intergenic module replacement have occurred between B. subtilis lipopeptide synthetases including the iturin family and the plipastatin and fengycin family. PMID- 15109719 TI - Genetic evidence for histidine kinase HP165 being an acid sensor of Helicobacter pylori. AB - Helicobacter pylori is a human gastric pathogen which is extremely well adapted to its unique habitat. Crucial for the survival under the acidic conditions prevailing in the stomach is the enzyme urease. Transcriptome analysis has shown that transcription of a large number of genes responds to the exposure of H. pylori to acid including the genes encoding the urease subunits UreA and UreB as well as several genes which have been previously identified as target genes of the two-component system HP166-HP165. Here, we provide genetic evidence that a stimulus perceived by the histidine kinase HP165 is pH since increased transcription at acidic pH from the promoters controlling the expression of the ORFs hp119 and hp1432 which belong to the HP166-HP165 regulon is strictly dependent on the presence of histidine kinase HP165. Furthermore, we show that the basal transcription from the promoter of the ureA gene is modulated by the HP166-HP165 two-component system in response to acidic pH. On the other hand, the acid-induced increase in transcription of the promoter directing the expression of the orphan response regulator HP1021 is not controlled by the HP166-HP165 two component system, nor is it mediated by HP1021 itself. PMID- 15109720 TI - Identification of a Shiga-toxin type I variant containing an IS1203-like element, from Shiga-toxin producing Escherichia coli O157:H7. AB - We found two Shiga toxin producing Escherichia coli O157:H7 strains isolated from humans carrying the stx(1) gene with an IS1203-like element (designated as IS1203v(1)). The IS1203v(1) was inserted into the coding region of the A subunit 7 bp upstream from the TGA termination codon, resulting in a loss of two amino acid residues (Ser-Ser) from its C terminus. Toxicity of the Stx1 was confirmed by Vero cell assay. IS1203v(1) hardly affected the stx(1) gene in either its expression or the toxicity of its product. PMID- 15109721 TI - Phytomonas iron superoxide dismutase: a possible molecular marker. AB - We have isolated and biochemically characterized two iron superoxide dismutases activities (SODI and SODII) from a plant trypanosomatid isolated from Euphorbia characias. The isoenzyme FeSODII has immunogenic capacity, and the positivity of the anti-SODII serum persists to a dilution of 1/40,000, by Western blot. In addition, Western blot has been used to test the positivity of the anti-SODII serum against antigen fractions (SOD) from 17 isolates belonging to the family Trypanosomatidae and for which we had previously determined the isoenzymatic profile. The reaction proved positive only with those plant isolates considered to belong to the genus Phytomonas, whereas there was no reaction of the anti SODII serum, against the antigen fractions from the species Trypanosoma cruzi, Leishmania donovani, Herpetomonas samuelpessoai, Herpetomonas davidi, Crithidia luciliae and Leptomonas collosoma. FeSODII is located mainly over the entire surface of the parasite, as well as in the nucleus, glycosomes and membranes. The above makes FeSODII promising as a molecular tool for diagnosis and identification, and as a potential chemotherapeutic target for designing drugs aimed at controlling not only of the diseases caused by Phytomonas species, but also for the great metabolic similarity to other trypanosomatids of animals and humans, it may be possible for these results to be extrapolated. Moreover, the sequencing of the amino-terminal end of the FeSODII enables the design of primers that in the near future will make it possible to sequence the gene of this isoenzyme. PMID- 15109722 TI - Novel pollutant-resistant methylotrophic bacteria for use in bioremediation. AB - Thirty one novel methylotrophic bacterial strains were isolated from a range of soil and sediment sources (both pristine and polluted) under different enrichment regimes. They were characterised physiologically and classified by their 16S rRNA gene sequence. A great taxonomical and phenotypical variety was recovered. Some of the isolates display interesting features of resistance to heavy metals, arsenate or organic pollution and four can be considered real 'super-bugs' for their ability to withstand extremely high concentrations of a variety of pollutants. A description of the 31 strains is presented in this work. PMID- 15109723 TI - Actinobacillus pleuropneumoniae metalloprotease: cloning and in vivo expression. AB - The complete amino acid and nucleotide sequence of a secreted metalloprotease produced by Actinobacillus pleuropneumoniae serotype 1 is reported. A clone showing proteolytic activity in cell-free culture media was selected from a genomic library of A. pleuropneumoniae serotype 1 in pUC 19. The sequence obtained contained an open reading frame encoding a protein with 869 amino acids. This protein was identified as a zinc neutral-metalloprotease belonging to the aminopeptidase family, with a predicted molecular weight of approximately 101 kDa. This sequence showed high homology with other predicted or sequenced aminopeptidases reported for different Gram-negative bacteria. Expression of the protease was observed in lung tissue from pigs that died of porcine pleuropneumonia suggesting a role in pathogenesis. PMID- 15109724 TI - ADP-ribosylation by exoenzyme T of Pseudomonas aeruginosa induces an irreversible effect on the host cell cytoskeleton in vivo. AB - Pseudomonas aeruginosa utilises a type III secretion system (TTSS) to introduce exoenzyme S and exoenzyme T into host cells to subvert host cell signalling and thereby promote infection. In this study, we have employed the heterologous TTSS of Yersinia to deliver different mutants of ExoT into HeLa cells. Wild-type ExoT and ExoT variants expressing either GAP (GTPase activating protein) or ADP ribosyltransferase activity mediated changes in cell morphology, which correlated to disruption of the actin microfilaments of the infected cells. ExoT expressing ADP-ribosylating activity gave an irreversible effect on HeLa cell morphology, while ExoT expressing only GAP activity displayed a reversible effect where the cells regained normal cell morphology after killing of the infecting bacteria. This shows that ExoT can modify and inactivate host cell proteins involved in maintaining the actin cytoskeleton in vivo by two independent mechanisms. PMID- 15109725 TI - Detection of gp43 of Paracoccidioides brasiliensis by the loop-mediated isothermal amplification (LAMP) method. AB - Paracoccidioidomycosis is a deep mycosis caused by the thermo-dependent dimorphic fungus Paracoccidioides brasiliensis and is prevalent in Latin American countries. We detected the species specific gp43 gene of P. brasiliensis by loop mediated isothermal amplification (LAMP) in 22 clinical and seven armadillo derived isolates. The amplified DNA appeared as a ladder with a specific banding pattern. The advantage of the LAMP method is speed; only 3 h were necessary for identification of the organism and diagnosis of the disease. We were also able to obtain positive results from DNA extracted from a paraffin-embedded tissue sample of paracoccidioidomycosis, suggesting that this method may achieve clinical application in the near future. PMID- 15109726 TI - Exploring the effectiveness of tazobactam against ceftazidime resistant Escherichia coli: insights from the comparison between susceptibility testing and beta-lactamase inhibition. AB - Thirteen clinical isolates of Escherichia coli resistant to ceftazidime that possessed an AmpC and other (beta-lactamases were identified. The effectiveness of different formulations of piperacillin/tazobactam to other beta-lactams was compared. Antibiotic susceptibility testing, polymerase chain reaction, amplification of blaTEM, blaSHV and blaAmpC, and enzyme-linked immunosorbent assays to identify AmpC beta-lactamases were performed. Hydrolysis rates were obtained and residual enzymatic activity was determined. Cefepime and ertapenem were more active than piperacillin/tazobactam. In contrast, increasing the relative proportion of tazobactam improved susceptibility testing. Twenty micromolar tazobactam inhibited total beta-lactamase activity (as measured by nitrocefin hydrolysis rates) by greater than 75% against all isolates tested: in 11 of 13 E. coli isolates, total beta-lactamase activity was inhibited by 90%. The observed differences between MIC determinations and susceptibility to enzymatic inactivation by tazobactam against E. coli containing AmpC and other lactamases may be due to the final tazobactam concentration achieved in the periplasmic space. Factors determining this are critical considerations in assessing beta-lactamase inhibitor potency. PMID- 15109727 TI - Identification and characterization of a novel inulin binding module (IBM) from the CFTase of Bacillus macerans CFC1. AB - A novel inulin-binding module (IBM), which was identified from the N-terminal region of the cycloinulinooligosaccharide fructanotransferase (CFTase) in Bacillus macerans CFC1, was characterized using the discrete entity of IBM produced by the recombinant Escherichia coli strains. Deletion analyses located the inulin binding activity in the N-terminal region between 241 and 389 amino acid residues, which was removed from the mature enzyme by processing when secreted from the B. macerans CFC1 cells. IBM bound specifically to polyfructans such as inulin and levan but it did not interact with any of the glycan polymers tested in this study including cellulose, xylan, and starch. Binding studies on the IBM revealed that the equilibrium dissociation constant K(d) and the maximum amount of protein bound [(PC)(max)] were 4.7 microM and 22 microM g(-1), respectively. Together, these results indicate that the IBM of CFTase has a relatively high and specific affinity for inulin. Adsorption of the IBM to inulin was highest at pH 7.0 and lowered slowly with decreasing pH down to 3.0. At pH 7.0, the binding activity was enhanced about twofold by the presence of 1 M MgCl(2). Chemical modification experiments with the aromatic amino acid-specific modifiers implied that tryptophan and tyrosine residues in the IBM are likely to participate in the interaction with the inulin molecules. PMID- 15109728 TI - Metabolomic differential display analysis of the white-rot basidiomycete Phanerochaete chrysosporium grown under air and 100% oxygen. AB - When the cultural atmosphere of the white-rot basidiomycete, Phanerochaete chrysosporium, was changed from air to 100% oxygen, the lyophilized mycelial weight increased and thickening of extracellular glucan layer was observed in 2-3 days. To better understand the oxygen-stress responsive mechanism of P. chrysosporium, the metabolomic differential display analysis was performed using metabolites isolated from fungal cells grown under either air or 100% O(2) atmosphere. In the GC-MS total ion chromatogram of methanol-extracts from fungal cells, at least 183 peaks were detected and 53 compounds were identified. Among them, veratryl alcohol (VA), threonate, and erythronate were identified as oxygen stress responsive metabolites. The intracellular concentration of VA increased dramatically within 1 h after an oxygen purge, indicating that VA production is sensitive to the oxygen stress in P. chrysosporium. PMID- 15109729 TI - The pentose phosphate pathway in Trypanosoma cruzi. AB - The pentose phosphate pathway has been studied in Trypanosoma cruzi, Clone CL Brener. Functioning of the pathway was demonstrated in epimastigotes by measuring the evolution of (14)CO(2) from [1-(14)C] or [6-(14)C]D-glucose. Glucose consumption through the PPP increased from 9.9% to 20.4% in the presence of methylene blue, which mimics oxidative stress. All the enzymes of the PPP are present in the four major developmental stages of the parasite. Subcellular localisation experiments suggested that the PPP enzymes have a cytosolic component, predominant in most cases, although all of them also seem to have organellar localisation(s). PMID- 15109730 TI - The O-antigen gene cluster of Shigella boydii O11 and functional identification of its wzy gene. AB - Shigella strains are human pathogens and their identification is usually based on their O-antigens. The O-antigen gene cluster of Shigella boydii O11 was sequenced. All the expected genes for the synthesis of the O-antigen were identified on the basis of homology and genes for the biosynthesis of dTDP-l Rhamnose, genes encoding sugar transferases, as well as genes encoding O unit flippase (wzx) and O-antigen polymerase (wzy). The identity of the putative wzy gene was confirmed by showing that a wzy deficient mutant strain of S. boydii O11 produced a semi-rough LPS phenotype. The predicted wzx gene has an opposite transcription direction to that of all of the other genes in the S. boydii O11 O antigen gene cluster. This unusual feature for the wzx gene has only previously been reported in S. boydii O6. Further comparison revealed an evolutionary relationship between O6 and O11 O-antigen gene clusters. Adjacent-gene PCR showed that Escherichia coli O105 and S. boydii O11, which share the identical O antigen, also have the same genes and organization for their respective O-antigen gene clusters. Three genes specific for the S. boydii O11 and E. coli O105 gene clusters were identified. PMID- 15109731 TI - A FhuA mutant of Escherichia coli is infected by phage T1-independent of TonB. AB - Infection of Escherichia coli K-12 by phages T1 and phi 80 requires the FhuA outer membrane protein and the TonB protein. Mutations in the N-terminal globular domain close to the predicted channel in the beta-barrel of FhuA were created. The FhuA Delta 107-111 N104K K110D L111P mutant and the FhuA(L(109)DPNGLK(110)) insertion mutant were sensitive to phage T1, but nearly resistant to phage phi 80. FhuA Delta 107-111 N104K K110D L111P mediated phage T1 infection in a tonB mutant without formation of TonB-independent phage T1 host-range mutants. The FhuA mutants showed no altered sensitivity to phage T5. Although the phages share overlapping binding sites in FhuA, the structural alterations elicited by the mutations resulted in very different phage sensitivities. In the FhuA deletion mutant, the TonB requirement for phage T1 infection was partially bypassed. PMID- 15109732 TI - Comparative analysis of transcriptional regulatory elements of glutamate dependent acid-resistance systems of Shigella flexneri and Escherichia coli O157:H7. AB - The ability to withstand an acid-challenge of pH 2.5 or less by Shigella flexneri is a necessary trait for virulence and is generally believed to be restricted to the stationary-phase of growth. Earlier reports indicated the glutamate-dependent acid-resistance (GDAR) system of S. flexneri is under the regulation of rpoS, the gene encoding alternative sigma factor that is induced in the stationary-growth phase. The present study reports that unlike Escherichia coli O157:H7, S. flexneri cells when grown in minimal medium, require acid-induction in the stationary-growth phase for a functional GDAR. When grown on complex medium at pH 5.5, GDAR of S. flexneri was vigorous compared to the cells grown at pH 7.5. No acid-induction was required for the stationary phase E. coli cells grown on either minimal or complex growth media. Distinct differences in the gadA, gadBC, gadE, and hdeA (but not in rpoS) transcript levels were observed in the stationary-growth phase cells between the two pathogens grown on minimal medium. Additionally, rpoS-independent acid-induction of GDAR in the logarithmic growth phase that has been recently observed in E. coli strains [FEMS Microbiol. Lett. 227 (2003) 39-45] was not detected in the S. flexneri rpoS mutant. Although some differences in the DNA sequence at the upstream regulatory elements of gadBC were noticed, they do not appear to be significant and involvement of additional regulators in S. flexneri is anticipated, which also may explain the observed differences in the GDAR of two enteric pathogens. PMID- 15109733 TI - Tyramine functions as a toxin in honey bee larvae during Varroa-transmitted infection by Melissococcus pluton. AB - From wounds of honey bee pupae, caused by the mite Varroa destructor, coccoid bacteria were isolated and identified as Melissococcus pluton. The bacterial isolate was grown anaerobically in sorbitol medium to produce a toxic compound that was purified on XAD columns, gelfiltration and preparative HPLC. The toxic agent was identified by GC-MS and FTICR-MS as tyramine. The toxicity of the isolated tyramine was tested by a novel mobility test using the protozoon Stylonychia lemnae. A concentration of 0.2 mg/ml led to immediate inhibition of mobility. In addition the toxicity was studied on honey bee larvae by feeding tyramine/water mixtures added to the larval jelly. The lethal dosis of tyramine on 4-5 days old bee larvae was determined as 0.3 mg/larvae when added as a volume of 20 microl to the larval food in brood cells. Several other biogenic amines, such as phenylethylamine, histamine, spermine, cadaverine, putrescine and trimethylamine, were tested as their hydrochloric salts for comparison and were found to be inhibitory in the Stylonychia mobility test at similar concentrations. A quantitative hemolysis test with human red blood cells revealed that tyramine and histamine showed the highest membranolytic activity, followed by the phenylethylamine, trimethylamine and spermine, while the linear diamines, cadaverine and putrescine, showed a significantly lower hemolysis when calculated on a molar amine basis. The results indicate that tyramine which is a characteristic amine produced by M. pluton in culture, is the causative agent of the observed toxic symptoms in bee larvae. Thus this disease, known as European foulbrood, is possibly an infection transmitted by the Varroa destructor mite. PMID- 15109734 TI - A von Willebrand factor-binding protein from Staphylococcus lugdunensis. AB - In the present study, a phage display library covering the genome of Staphylococcus lugdunensis, was affinity-selected against von Willebrand factor (vWf). This led to the identification of a gene, vwbl, encoding a putative cell surface protein of 2060 amino acids, denoted vWbl. The deduced protein has an overall organisation typical of staphylococcal cell surface proteins, with an N terminal signal peptide, and a C-terminal cell wall sorting signal. The vWf binding part is located in repetitive domains and antibodies against vWbl or vWf can inhibit the binding. Southern blot analysis showed that vwbl was present in the 12 S. lugdunensis strains tested. PMID- 15109735 TI - Significance of Na+ in the fish pathogen, Vibrio anguillarum, under energy depleted condition. AB - Vibrio anguillarum kills various kinds of fish over salinities ranging from seawater to freshwater. In this study, we investigated the role of Na(+) in V. anguillarum, especially under energy-depleted conditions such as in natural seawater. V. angustum S14, which is a typical marine vibrio, was used for comparison. V. anguillarum only required Na(+) for starvation-survival, but in contrast, V. angustum S14 always required Na(+) for both growth and starvation survival. In marine vibrios, Na(+) is used in the Na(+)-dependent respiratory chain that produces the sodium motive force (SMF) across the cell membrane. It has been considered that marine vibrios always need a SMF produced by Na(+), however in the case of V. anguillarum, the SMF is not required for growth, but becomes more important for starvation-survival. PMID- 15109736 TI - Mineralization of erythromycin A in aquaculture sediments. AB - Mineralization of erythromycin A was studied using two differently (14)C-labeled erythromycins A, which were added to aquaculture sediment samples obtained from the two salmon hatchery sites in Washington state. The added erythromycin A did not significantly alter the numbers of the total viable colonies and erythromycin resistant bacteria. Erythromycin-resistant Pseudomonas species contained a constitutive erythromycin esterase activity contributing to the inactivation of biologically active erythromycin A in aquatic and sediment environments. The initial rate of mineralization of erythromycin A appeared to be governed by the rate of release of soil-sorbed erythromycin A. After a prolonged lag time, the S curves of erythromycin A mineralization were observed probably because of the increase in the population density metabolizing it. This study suggests that erythromycin A is partially or completely mineralized by the sediment microbial populations. PMID- 15109737 TI - Production of an antifungal protein for control of Colletotrichum lagenarium by Bacillus amyloliquefaciens MET0908. AB - A plant pathogenic fungus, Colletotrichum lagenarium, causing watermelon anthracnose, was isolated from naturally infected leaves, stems, and fruits of watermelon. A bacterial strain, MET0908, showing a potent antifungal activity against C. lagenarium, was isolated from soil. An antifungal protein was purified by 30% ammonium sulfate saturation and concentrated using Centricon 10, DEAE Sepharose(TM) Fast Flow column and Sephacryl S-100 gel filtration chromatography. The molecular weight of the purified protein was estimated as 40 kDa by SDS-PAGE. The purified protein was stable at 80 degrees C for 20 min and exhibited a broad spectrum of antifungal activity against various plant pathogenic fungi. Confocal microscopy image analysis and scanning electron microscopy showed that the protein acted on the cell wall of C. lagenarium. The purified antifungal protein exhibited beta-1,3-glucanase activity. The N-terminal amino acid sequence of the purified protein was determined as Ser-Lys-Ile-x-Ile-Asn-Ile-Asn-Ile-x-Gln-Ala Pro-Ala-Pro-x-Ala. A search of the sequence with NCBI BLAST showed no significant homology with any known proteins, suggesting that the purified protein may be novel. PMID- 15109738 TI - The AMPPA network as a successful model for public-private or private-private partnership. AB - In 1997, the Rockefeller Foundation and the Ernst Schering Research Foundation (a subsidiary of Schering AG, Germany, on a non-profit basis) mounted a multi-year global collaborative effort, involving a network of top-level research institutions to intensify research on the regulation of the male reproductive system with special emphasis on post-testicular activity, utilizing new approaches in molecular pharmacology (application of molecular pharmacology for post-testicular activity (AMPPA) network). The new venture proved a success as a public-private sector partnership, as a collaborative scientific program, and as an approach to identify new targets applicable and suitable for drug finding for male fertility regulation. PMID- 15109739 TI - Regulation of epididymal principal cell functions by basal cells: role of transient receptor potential (Trp) proteins and cyclooxygenase-1 (COX-1). AB - The epithelia lining the epididymides of many species including the human are known to consist of several cell types. Among them, the principal cells are the most abundant and their functions most extensively studied. There are other cell types such as the narrow cells, clear cells, halo cells and basal cells which are scattered along the duct in lesser number. Although these minority cell types have not been studied to the same extent as the principal cells, it is conceivable that their presence are essential to the integrated functions of the epididymis. In the intact epididymis, basal cells can be seen adhering to the basement membrane forming close contact with the principal cells above them. Work in our laboratory has provided evidence that through local formation of prostaglandins, basal cells may regulate electrolyte and water transport by the principal cells. This regulatory process involves two proteins which are exclusively expressed by the basal cells. They are the transient receptor potential (Trp) proteins, which serve as transmembrane pathways for Ca(2+) influx, and cyclooxygenase 1 (COX-1), a key enzyme in the formation of prostaglandins. The role of the two proteins in the integrated functions of the basal cells as humoral regulators of principal cells is discussed. PMID- 15109740 TI - Epididymis-specific promoter-driven gene targeting: a new approach to control epididymal function? PMID- 15109741 TI - Technologies for the study of epididymal-specific genes. AB - Sperm maturation occurs during transit through the epididymis. Interactions between the epididymal epithelium and the sperm are crucial for the maturation process. Analyses of existing male-infertile mouse lines have begun to enumerate some of the genes involved. Recent advances in transgenic technologies to generate temporally and spatially restricted targeted gene disruptions show promise for progress in understanding sperm maturation. Gene silencing agents, such as RNAi, to manipulate gene expression may prove useful for the analysis of epididymal genes involved in the maturation process. PMID- 15109742 TI - Antioxidant strategies in the epididymis. AB - Spermatozoa are very specialized cells, dedicated to fertilization of the oocyte. The attainment of this biological role is partly due to the fusogenic properties of the sperm plasma membrane, which is particularly rich in polyunsaturated fatty acids (PUFA). This predominance of PUFA renders spermatozoa highly susceptible to lipid peroxidation due to attacks from reactive oxygen species (ROS). These attacks ultimately lead to the impairment of sperm function through oxidative stress. Despite such disruptive effects, it should be also emphasized that these molecules also play an important positive, physiological role in the regulation of sperm physiology through their participation in apoptosis and the signal transduction cascades that control sperm maturation and capacitation. In this article, the different sources of ROS are examined and then the antioxidant strategies that protect these cells during epididymal transit are reviewed. While the major focus is on the involvement of glutathione peroxidase in this process, consideration will also be given to a range of additional antioxidant enzymes (catalase, indolamine dioxygenase and superoxide dismutase) that have evolved to protect spermatozoa during this extremely vulnerable phase in their life history. Besides the classical enzymatic roles of these enzymes in recycling ROS, additional features are discussed in the light of contraceptive development. PMID- 15109743 TI - Molecular diffusion in sperm plasma membranes during epididymal maturation. AB - Fluorescence recovery after photobleaching (FRAP) analysis has been used to measure lipid diffusion in different regions of the sperm plasma membrane. Our goal has been to understand how some membrane components are confined to specific surface domains, whilst others are freely diffusing and in some cases are able to migrate against large concentration gradients. Results with a variety of fluorescent lipid reporter probes (ODAF, NBD-PC, NBD-cholesterol) show that diffusion coefficients (D) are generally three to four times higher on the sperm acrosome than on the principal piece of the tail and increase significantly during epididymal maturation (ram, mouse, goat, dog and monkey sperm). Cholesterol diffusion is approximately 10 times faster on the sperm head than the tail and has a heterogenous distribution when detected with filipin. Lipid diffusion is very temperature sensitive but remarkably insensitive to changes in external pH and osmotic pressure. There was no evidence that the posterior ring or annulus functioned as diffusion barriers to lipids. On this basis it was possible to construct models of increasing complexity to describe the behaviour of a lipid molecule on the sperm surface, beginning with simple linear diffusion progressing to random diffusion and eventually to constrained diffusion. PMID- 15109744 TI - The importance of redox regulated pathways in sperm cell biology. AB - Redox regulated events are fundamental to our understanding of many cellular pathways and pathological processes. On the one hand, production of reactive oxygen species by mammalian spermatozoa has been associated with a loss of cell function and DNA integrity as a consequence of oxidative stress. These cells are exquisitely sensitive to such damage as a consequence of their relative lack of cytosolic antioxidant enzymes and relative abundance of polyunsaturated fatty acids. Given this susceptibility, it is surprising to discover that spermatozoa are intensely redox active cells and professional generators of reactive oxygen species. The latter are physiologically important to the spermatozoa in regulating every aspect of sperm function examined, including their movement characteristics, capacitation, sperm-zona interaction, the acrosome reaction and sperm-oocyte fusion. The molecular basis of this redox drive is still poorly understood in terms of the source of the reactive oxygen species and the mechanisms by which these reactive metabolites enhance sperm function. Recent advances include the discovery of NOX5 in the male germ line and elucidation of the role of reactive oxygen species in controlling a unique signal transduction cascade associated with sperm capacitation. Given the central importance of redox chemistry in the control of sperm function further research in this area may uncover valuable targets for contraceptive intervention. PMID- 15109745 TI - Mouse models of infertility due to swollen spermatozoa. AB - Transgenic mice with male infertility, the c-ros knockout (KO) and GPX5-Tag2 transgenic mouse models, are compared. Both exhibit severely angulated sperm flagella explaining the infertility. As angulated spermatozoa are swollen cells, a failure in volume regulation is indicated. Differences between genotypes were also found: caudal spermatozoa from c-ros KO, but not GPX5-Tag2, could fertilise eggs in vitro; flagellar angulation occurred more within the epididymis of GPX5 Tag2 than c-ros KO mice; the osmotic pressure of cauda epididymidal fluid was lower only in GPX5-Tag2 mice; angulation of caudal sperm from c-ros KO, but not GPX5-Tag2 mice, decreased upon demembranation. These observations indicate that GPX5-Tag2 mice express an earlier, more severe defect. Gene chip analyses of the epididymides revealed decreased expression of the CRES (cystatin-related epididymal-spermatogenic) and MEP17 (murine epididymal protein 17) genes in both genotypes. Further analysis could pinpoint genes essential for epididymal regulation of sperm volume, explain infertility and suggest modes of male contraception. PMID- 15109746 TI - Post-meiotic gene products as targets for male contraception. AB - Post-meiotic stages of male germ cell maturation represent an interesting target system for the development of novel male contraceptive agents. In the human, these stages represent a period of only about 16 days differentiation, and thus targeting these cells would represent a contraceptive approach with a relatively rapid onset and equivalent recovery. Results from the Human Genome Project suggest that these cells also express a high number of very specific transcripts, though whether all of these are functional and/or essential for sperm differentiation and function requires more research. Until recently, however, these haploid stages were relatively inaccessible to molecular research because of the lack of appropriate model systems and methods. This situation has recently improved, with several new techniques involving manipulation of primary cells and seminiferous tubules, germ cell transplantation and the development of new immortalized cell-lines. Also, new biochemical approaches are yielding more information about haploid-specific transcription factors, such as GCNF. It is therefore to be expected that soon several new targets for a potential post meiotic male contraceptive will become available for pharmaceutical development. PMID- 15109747 TI - Male contraception-a topic with many facets. AB - In addition to the scientific issues associated with male contraception, there are a variety of other concerns that must be addressed before new male contraceptives reach the market. Cultural attitudes toward contraception will play a role both in the acceptability of any contraceptive and in compliance and usage. Delivery methods must also be considered; different methods are favored depending on the social context. Prevention of sexually transmitted diseases by a combined contraceptive/microbicidal treatment is a laudable goal, and may enhance public acceptance of a male contraceptive. This review is the result of a workshop that was convened to address these topics. PMID- 15109749 TI - Male contraception, a choice on the horizon. Proceedings of the final meeting of the joint Rockefeller/Ernst Schering Network on Epididymal Research (AMPPA: Applied Molecular Pharmacology of Posttesticular Activity). October 2002. Bellagio Italy. PMID- 15109748 TI - Immortalization by large T-antigen of the adult epididymal duct epithelium. AB - The SV40 large T-antigen has been widely used to convert various cell types to a transformed phenotype, and also to induce progressive tumours in transgenic animals. The objectives of this review are to compare and discuss three different approaches to generate epididymal epithelial cell lines using the large T antigen. In the first approach, retroviral transfection of primary cultures was used to immortalize canine epididymal cells in vitro; the other two approaches used transgenic mice expressing the large T-antigen. In one of these in vivo approaches, a construct consisting of the coding sequence of a temperature sensitive (ts) SV40 large T-antigen was inserted in a mouse genome. When the cells are exposed to the permissive temperature of 33 degrees C, functional expression of the large T-antigen occurs and cells start to proliferate. In the second in vivo approach a tissue-specific promoter, the 5kb GPX5 promoter, was used to direct expression of the large T-antigen to the epididymal duct epithelium. PMID- 15109750 TI - The influence of slouching and lumbar support on iliolumbar ligaments, intervertebral discs and sacroiliac joints. AB - OBJECTIVE: To investigate lumbopelvic kinematics when moving into a slouch. DESIGN: A biomechanical model was developed. Load tests in vitro verified the model. BACKGROUND: The precise mechanism causing disc herniation and back sprain is still debated. Most biomechanical studies have focused on lifting in a stooped posture. Previous studies address instability situations due to Euler buckling of the spine under axial load. However, no studies address lumbosacral, iliolumbar and sacroiliac kinematics in slouching, i.e. flexing the spine in situations with negligible compressive spinal load. METHODS: Modeling started with the click clack movement, i.e. the transition from lumbar lordosis to lumbar kyphosis by the combination of backward rotation of the pelvis and ventral flexion of the spine. The flexed spine was compared with a crowbar which uses the iliolumbar ligaments as fulcrum and pivot. To analyse the click-clack movement in sitting, unembalmed erect human trunks were moved from a forward position to a backward position, recording angular changes between L5, sacrum and ilium. RESULTS: When moving the trunk stepwise backward with support at shoulder level, L5 showed forward rotation with respect to the sacrum, but rotation of the sacrum with respect to the iliac bones was reversed (i.e. counternutation). L5 showed displacement in ventral direction with respect to the ilium. Measurements were in agreement with prediction from the crowbar model of the spine. CONCLUSIONS: Backward rotation of the pelvis combined with flexion of the spine, i.e. slouching, results in backward rotation of the sacrum with respect to the ilium, dorsal widening of the intervertebral disc L5-S1 and strain on the iliolumbar ligaments when protection from back muscles against lumbar flexion is absent. Lumbar backrest support almost eliminates lumbosacral and sacroiliac movement. RELEVANCE: Understanding why the iliolumbar ligaments are loaded in slouching contributes to the understanding of the biomechanics of low back pain in everyday situations with small or negligible compressive spinal load. The results recommend lumbar support: backrests with free shoulder space. PMID- 15109751 TI - The time-varying response of the in vivo lumbar spine to dynamic repetitive flexion. AB - OBJECTIVE: To quantify the time-varying stiffness and kinematic responses of the in vivo lumbar spine exposed to dynamic repetitive flexion movements. DESIGN: Changes in in vivo passive lumbar moment-angle relationships were monitored in response to dynamic repetitive flexion. BACKGROUND: While previous in vitro studies have provided conflicting evidence on the effects of repetitive flexion movements on the stiffness of the lumbar spine, no previous studies have quantified the time-varying changes of the in vivo lumbar spine to dynamic repetitive flexion. METHODS: Subjects lifted and lowered a 4.5 kg load over two barriers at a rate of 7 lifts per minute for 1.5 h inducing at least 80% of the lumbar flexion range of motion. Prior to lifting and at 30 min intervals passive moment-angle relationships were obtained by pulling the subject into flexion on a customized frictionless table. RESULTS: Repetitive lifting induced a decreasing stiffness trend after 30 min, followed by a recovery towards initial stiffness levels with further loading. The trends were non-significant for all measures studied. CONCLUSIONS: The results indicate that after 30 min of lifting, creep within the passive tissues may allow workers to exceed their initial range of motion, altering joint mechanics and loading patterns potentially leading to an increased risk of developing low back pain. RELEVANCE: Given the potential for time-varying changes to alter the spine's risk of injury and injury mechanisms, knowledge regarding the stiffness response of the in vivo lumbar spine exposed to repetitive flexion may lead to improved understanding and prevention of work related back pain. PMID- 15109752 TI - The relationship between EMG and change in thickness of transversus abdominis. AB - OBJECTIVE: To investigate the relationship between changes in thickness and EMG activity in the transversus abdominis muscle of healthy subjects and the reliability of ultrasound measurements using different modes and transducers. DESIGN: Convenience sampling. BACKGROUND: Chronic low back pain is associated with transversus abdominis dysfunction but EMG studies of this muscle are restricted to invasive techniques. Since the thickness of transversus abdominis changes with activity, such changes measured from ultrasound images might provide insight into this muscle's function non-invasively. In addition, little is known about the comparability of ultrasound measurements from different modes and transducers, nor the reliability of transversus abdominis measurements. METHODS: In 9 healthy subjects (aged 29-52 years, four male) transversus abdominis was studied at rest and during activity (5-80% max) with simultaneous EMG and ultrasound (M mode, 5 MHz curvilinear transducer) measurements. Intra-rater reliability for thickness measurements was studied on 13 subjects using 7.5 MHz linear and 5 MHz curvilinear transducers in B and M modes. RESULTS: Muscle thickness changes correlated well with EMG activity (P < 0.001, R2 = 0.87) and there were no significant differences between subjects (P > 0.05). Using 7.5 MHz head, the ICC for B mode was 0.989 and for M mode was 0.981 for between days reliability. The ICC for between transducer reliability was 0.817. CONCLUSIONS: Changes in thickness of transversus abdominis can be used to indicate changes in the electrical activity in this muscle. RELEVANCE: Ultrasound scanning can be used in the clinical setting to provide objective information about transversus abdominis function. PMID- 15109753 TI - Mechanical efficiency during hand-rim wheelchair propulsion: effects of base-line subtraction and power output. AB - OBJECTIVE: The purpose of this study was to compare four methods of calculating the mechanical efficiency of hand-rim wheelchair propulsion. METHODS: After completing a maximal incremental test, 18 untrained able-bodied males (mean age 22 years) performed four 4-min bouts of exercise (0%, 40%, 55%, and 70% of peak power output) on a wheelchair-ergometer. The mechanical efficiency was calculated during each submaximal bout by the ratio of work accomplished to the amount of energy expended with four indices: gross (no base-line correction), net (resting metabolism as base-line correction), work (unloaded movement as base-line correction) and delta (measurable work as base-line correction) indices. RESULTS: Work efficiency (16.8-13.8%) were significantly higher (P < 0.001) than gross (6.5-8.9%) or net efficiency (9.9-11.1%) at 40-70% peak power output, respectively. Net efficiency was also significantly lower than work efficiency (P < 0.001) at all intensity. Moreover, gross and work efficiency increased and decreases (P < 0.001) respectively with intensity. In contrast, net and delta efficiency remained unchanged. CONCLUSIONS: Depending upon the base-line used, each efficiency index provided different values and were differently affected by the exercise intensity. Precautions should be taken on the conclusions about the mechanical efficiency of hand-rim wheelchair propulsion. Moreover, comparisons between the four efficiency indices among themselves allowed a better and complete description of wheelchair locomotion and wheelchair dependent person. Gross or net efficiencies better reflected the actual mechanical efficiency of the hand-rim wheelchair propulsion whereas work or delta efficiencies investigated the efficiency of muscular contraction during this movement. RELEVANCE: Since the four mechanical efficiency indices measured different aspects of performance, it will have consequences when it comes to choosing an appropriate index. The better understanding of mechanical efficiency index may help future investigators to choose the most appropriate index for their purposes. PMID- 15109754 TI - Biomechanical analysis of tendon transfers for massive rotator cuff tears. AB - OBJECTIVE: To determine why certain tendon transfers are mechanically more effective than other tendon transfers for the treatment of a massive rotator cuff tear. DESIGN: A tendon transfer procedure of latissimus dorsi, teres major or a combination of these two to the insertions of either teres minor, infraspinatus, supraspinatus, or subscapularis is simulated using a biomechanical musculoskeletal model of the upper extremity. BACKGROUND: Massive rotator cuff tears are not easily repaired. To compensate for the loss of rotator cuff function, techniques such as muscle transfers are developed. METHODS: Three range of motion tasks were used as input to the Delft shoulder and elbow model. The muscle parameters of the Delft shoulder and elbow model were modified to simulate a rotator cuff tear. A biomechanical analysis of the transferred muscles was performed, taking outcome variables such as moment arms, muscle length and muscle force into account. RESULTS: Due to the massive rotator cuff tear, an elevation and external rotation moment is lost. When the tendon was transferred to the insertions of infraspinatus or supraspinatus, the humerus was capable of elevating and externally rotating. CONCLUSIONS: On the basis of mechanical parameters such as moment arms, muscle length and force it can be concluded that a tendon transfer of the teres major to the supraspinatus insertion will produce the best functional outcome in the treatment of massive rotator cuff tears. RELEVANCE: To find biomechanical evidence for an optimal tendon transfer that will lead to improved treatment of patients with a massive rotator cuff tear. PMID- 15109755 TI - Internal fixation of radial neck fractures: an in vitro biomechanical analysis. AB - OBJECTIVE: To assess the fixation rigidities of a custom designed blade plate, a 2.7 mm T-plate and 3.0 mm cannulated screws. DESIGN: A cadaveric non-comminuted radial neck fracture model was employed to test the three fixation methods, using a multi-directional shear loading protocol. BACKGROUND: The management of displaced radial neck fractures in adults remains unsatisfactory due to failure of internal fixation with secondary loss of reduction, and non-unions. METHODS: The stiffness of the various fixation methods was measured in five directions of loading in the transverse plane. Failure testing was performed on the final testing condition for each specimen, in a posteroulnar to anteroradial direction. RESULTS: The direction of specimen loading did not have a significant effect on the stiffness of the various methods of fixation (P = 0.4). There was no significant difference in the fixation stiffness between the 3.0 mm screws or blade plate, however, both were superior to the T-plate (P < 0.05). CONCLUSIONS: Although the 2.7 mm T-plate has been reported in the literature as a viable method of fixation for radial neck fractures, it may not be optimal from the viewpoint of fixation stability. In the setting of a non-comminuted radial neck fracture, the use of cross-cannulated screws or blade plate fixation is preferred.Relevance This study supports the use of cross-cannulated screws or blade plate fixation for non-comminuted fractures of the radial neck. PMID- 15109756 TI - Appropriate radial clearance of ceramic-on-ceramic total hip prostheses to realize squeeze-film lubrication. AB - OBJECTIVE: Estimation of appropriate radial clearance of ceramic-on-ceramic total hip prostheses to realize squeeze-film lubrication. BACKGROUND: Some clinical results show that severe wear occurs at ceramic-on-ceramic interfaces of total hip prostheses if the design parameters are improper. Appropriate design is required to realize the optimum lubrication of joint prostheses. DESIGN: Squeeze film thickness of total hip prostheses with different radial clearances was numerically estimated under the physiological conditions of normal walking. METHODS: The changes in the fluid film thickness between ceramic balls and a ceramic cups with radial clearances of various values from 10 to 80 microm were numerically estimated under the assumption of normal walking conditions by means of elastohydrodynamic squeeze-film lubrication theory including the three dimensional theory of elasticity. RESULTS: The minimum film thickness remained over 0.02 microm during a few walking steps with a radial clearance under 20 microm and a head diameter of 28 mm. On the other hand, the fluid film was squeezed out during a few steps when the radial clearance was greater than 30 microm. CONCLUSIONS: Squeeze-film lubrication can be realized in a ceramic-on ceramic total hip prosthesis if the radial clearance is smaller than 20 microm on a head of 28 mm diameter. RELEVANCE: Fluid film lubrication is necessary to prevent the severe wear of ceramic joint prostheses. The present investigation shows which radial clearance of ceramic-on-ceramic hip prostheses is appropriate for squeeze-film lubrication under physiological conditions in daily action. PMID- 15109757 TI - Distribution of bone mineral density with age and gender in the proximal tibia. AB - OBJECTIVE: To investigate both the age and gender related distributions of bone mineral density in the proximal tibia, specifically in aged patients. BACKGROUND: For surgeons to achieve stable long-term fixation of implants in the proximal tibia, the distribution of bone mineral density must first be known. The changes that occur due to age or gender can alter these distributions. METHODS: Quantitative computed tomography and indentation testing were used to investigate 40 human tibiae (27 female, 13 male, average age 63.3 years). RESULTS: A significant reduction in bone mineral density was found in female tibiae between the age groups of < 60 and > or =60. This difference was not found in the male groups and no other significant difference was found between consecutive age groups. A three-dimensional map of the bone mineral density of the proximal tibia is therefore presented for the groups female < 60, female > or =60 and male. Reduced bone mineral density was consistently found in the central regions, whilst the regions of highest bone quality varied from postero-lateral to postero medial. CONCLUSIONS: Implant fixation for fracture treatment as well as joint replacement of the proximal tibia are now able to take the regions of both high and low bone mineral density into consideration in older patients and those suffering from osteoporosis. RELEVANCE: Knowledge regarding the regional distribution of bone mineral density in the proximal tibia is necessary in order to achieve stable primary and long-term fixation of implants. This manuscript documents the bone mineral density changes that occur with age and gender. PMID- 15109758 TI - Muscle activation and force production in Parkinson's patients during sit to stand transfers. AB - OBJECTIVE: The purpose of this study was to compare the kinematics, muscle activation, and force production between Parkinson's patients and healthy, age matched participants during sit-to-stand transfers. DESIGN: This cross-sectional study employed a 2x2x3 multivariate analysis of variance to test for significant differences between and within groups. BACKGROUND: The underlying mechanisms that predispose an individual to lose strength during the clinical progression of Parkinson's disease have proved to be elusive, especially during performance of functional tasks such as the sit to stand transfer. METHODS: Twenty-four men (mean age: 71.5 years) categorized as Parkinson's patients (n = 13) and healthy adults (n = 11) participated in this study. Two force platforms measured antero posterior and vertical force components as well as peak torque. Muscle activation was measured by a six channel, bilateral electromyography system. A lower-body kinematic assessment was conducted utilizing a high-speed motion analysis system. RESULTS: No statistically significant differences were found between groups for the outcome variables measured. However, Parkinson's patients did exhibit significant within-group bilateral differences for the variables of knee angle at seat-off, peak vertical force and peak torque. CONCLUSION: Data from this study reveal that persons with mild to moderate Parkinson's disease exhibit moderately altered bilateral mechanics when performing a sit to stand transfer compared to their healthy peers. RELEVANCE: The inability to produce constant equilateral force when performing functional tasks could be an indicator for the increased propensity of falls or other instabilities in this population. PMID- 15109759 TI - Defining phases for the sit-to-walk movement. AB - OBJECTIVE: To define phases of the sit-to-walk movement and test their consistency in a normal population. DESIGN: An observational study of thirteen healthy volunteers. BACKGROUND: Moving from sitting to walking is a daily activity that may present difficulty for some populations in terms of control and stability. METHODS: The movement was partitioned into phases according to changes in ground reaction forces and peak velocity of the total body centre of mass. Consistency of each phase duration was assessed. RESULTS: Four phases of sit-to walk were defined; flexion momentum, extension, unloading and stance. ICC scores for phase duration ranged from 0.54 (extension) to 0.81 (stance). CONCLUSIONS: This is the first study to define distinct phases of the sit-to-walk movement. There was moderate to good consistency for phase duration. RELEVANCE: By defining and testing phases of the sit-to-walk movement this study enhances understanding of this everyday movement and provide a model for future research. PMID- 15109760 TI - Mechanics and control of the flat versus normal foot during the stance phase of walking. AB - OBJECTIVE: To compare walking stance phase rearfoot and forefoot motion, ankle joint moments and extrinsic foot muscle EMG profiles between normal and pes planus feet. DESIGN: A cross-sectional comparative study. BACKGROUND: Musculoskeletal conditions are often attributed to pes planus, in which motion is assumed to be excessive and the muscle control inadequate. However, many of the speculated differences in mechanics and control between the normal and the pes planus foot have not been quantified. METHODS: Kinematic and kinetic data were obtained from video recordings of skin surface markers and a force plate, and EMG was recorded with surface electrodes. Analysis of variance was carried out to compare the group profiles. RESULTS: In the pes planus group: the forefoot was less adducted (P < .05) at toe-off, and total transverse plane range of motion, at 8 degrees versus 10 degrees, was less (P < .01); the peak plantarflexor ankle moment at push-off was greater (P < .05); the invertor moment was greater at foot flat (P < .05); for the EMG profiles, activity early in stance, relative to the mean stance phase activity was higher (P < .01) in tibialis anterior and lower (P < .05) in the peronei, soleus and medial and lateral gastrocnemius. CONCLUSIONS: Despite reaching statistical significance, the group differences were small for the task of laboratory walking at a natural pace. The main implications of the differences were for restraint of motion. The expectations of excessive motion and muscle effort in the pes planus group were therefore not substantiated. RELEVANCE: Symptomatic pes planus subjects did not reveal the expected biomechanical differences from normal subjects. The underlying causes of symptoms were not identifiable. PMID- 15109761 TI - Slow passive stretch and release characteristics of the calf muscles of older women with limited dorsiflexion range of motion. AB - OBJECTIVE: Examine the slow passive stretch and release characteristics of the calf muscles of older women with limited dorsiflexion range of motion. DESIGN: A cross-sectional comparative design. BACKGROUND: The passive stretch and release characteristics of the calf muscles of older women with limited dorsiflexion range of motion have not been studied. METHODS: Fifteen older women (mean 79 years) with active dorsiflexion < or =10 degrees and 15 younger women (mean 24 years) without limited dorsiflexion were tested. The right ankle was stretched from plantarflexion to maximal dorsiflexion and released into plantarflexion at 5 degrees /s with minimal surface EMG activity in the soleus, gastrocnemius, and tibialis anterior muscles. Length, passive-elastic stiffness and stored passive elastic energy were examined. RESULTS: The older women had less maximal passive dorsiflexion, a greater initial stretch angle, and less angular change than the younger women (P < 0.05). The maximal passive resistive force (Newtons) of the stretch phase, and the stored passive-elastic energy ( degrees N) during both stretch and release phases were also less (P < 0.001). The older women had greater passive-elastic stiffness at 0 degrees and 5 degrees of dorsiflexion (P < 0.001). CONCLUSIONS: The older women had decreased calf muscle length, extensibility, maximal passive resistive force, stored passive-elastic energy, but greater angle-specific-stiffness at 0 and 5 degrees of passive dorsiflexion. RELEVANCE: Older women with limited dorsiflexion range of motion have decreased calf muscle length, passive resistive forces and stored passive-elastic energy that may impact static and dynamic standing balance activities. Greater passive elastic stiffness within their ambulatory dorsiflexion range of motion may partially compensate for the deficits. PMID- 15109762 TI - Roll-over shapes of human locomotor systems: effects of walking speed. AB - OBJECTIVE: To examine the hypothesis that roll-over shapes of non-disabled lower limb systems do not change appreciably with walking speed. DESIGN: Repeated measures (n = 24). BACKGROUND: Roll-over shapes of three lower limb systems are presented. They are: roll-over shapes of the (1) foot, (2) ankle-foot, and (3) knee-ankle-foot systems. Roll-over shapes show the effective rocker (or cam) shapes that the lower limb systems conform to during the period in the stance phase of walking between heel contact and opposite heel contact. METHODS: Roll over shapes were measured by transforming center of pressure data from a laboratory-based coordinate system into each of three body-based coordinate systems. Knee-ankle-foot roll-over shapes were further characterized using a circular arc model. RESULTS: From a statistical standpoint, the radii of the best fit circular arcs did not change significantly with walking speed, while the forward shifts of the circular models did change significantly. However, the change in forward shift was not considered to be clinically significant. CONCLUSIONS: The biologic systems involved in developing the roll-over shapes adapt to changing conditions of walking speed, including increased loading amplitudes as speed is increased, to maintain similar effective roll-over geometries. RELEVANCE: Roll-over shapes provide insight into the workings of various lower limb systems by taking a new look at existing gait data. This insight could provide broad utility, helping to develop a better understanding of able-bodied and disabled human walking, and leading to the design of improved rehabilitation devices, surgeries, and therapies. PMID- 15109763 TI - PCA in studying coordination and variability: a tutorial. AB - OBJECTIVE: To explain and underscore the use of principal component analysis in clinical biomechanics as an expedient, unbiased means for reducing high dimensional data sets to a small number of modes or structures, as well as for teasing apart structural (invariant) and variable components in such data sets. DESIGN: The method is explained formally and then applied to both simulated and real (kinematic and electromyographic) data for didactical purposes, thus illustrating possible applications (and pitfalls) in the study of coordinated movement. BACKGROUND: In the sciences at large, principal component analysis is a well-known method to remove redundant information in multidimensional data sets by means of mode reduction. At present, principal component analysis is starting to penetrate the fundamental and clinical study of human movement, which amplifies the need for an accessible explanation of the method and its possibilities and limitations. Besides mode reduction, we discuss principal component analysis in its capacity as a data-driven filter, allowing for a separation of invariant and variant properties of coordination, which, arguably, is essential in studies of motor variability. METHODS: Principal component analysis is applied to kinematic and electromyographic time series obtained during treadmill walking by healthy humans. RESULTS: Common signal structures or modes are identified in the time series that turn out to be readily interpretable. In addition, the identified coherent modes are eliminated from the data, leaving a filtered, residual pattern from which useful information may be gleaned regarding motor variability. CONCLUSIONS: Principal component analysis allows for the detection of modes (information reduction) in both kinematic and electromyographic data sets, as well as for the separation of invariant structure and variance in those data sets. RELEVANCE: Principal component analysis can be successfully applied to movement data, both as feature extractor and as data driven filter. Its potential for the (clinical) study of human movement sciences (e.g., diagnostics and evaluation of interventions) is evident but still largely untapped. PMID- 15109764 TI - Restored flexor carpi ulnaris function after mere tenotomy explains the recurrence of spastic wrist deformity. AB - OBJECTIVE: To prove that fibrous restoration of the continuity of a cut tendon may cause recurrence of flexion deformity of the wrist after mere tenotomy of the spastic flexor carpi ulnaris muscle. BACKGROUND: Mere tenotomy of the flexor carpi ulnaris tendon is insufficient to prevent recurrence of acquired spastic flexion deformity of the wrist. Subsequent restoration of the continuity of the tendon by fibrous interposition may result in the recurrence. We examined whether a previously tenotomised muscle is strong enough to cause the deformity. METHODS: Active and passive force-length characteristics of the flexor carpi ulnaris muscle were measured intraoperatively in a patient with recurrent spastic flexion wrist deformity. The observed characteristics were compared with the average in vivo force-length characteristics of 14 spastic flexor carpi ulnaris muscles that had not previously been operated. RESULTS: The previously tenotomised flexor carpi ulnaris muscle was able to maximally exert 110 N force. Its active force length curve and passive force at maximal extension were similar to those of non operated spastic flexor carpi ulnaris muscles. CONCLUSIONS: A previously tenotomised flexor carpi ulnaris muscle is strong enough to cause recurrence of spastic flexion deformity of the wrist in case functional fibrous restoration of the tendon occurs after mere tenotomy. RELEVANCE: The surgical routine of mere tenotomy should probably be modified by including the dissection of the distal muscle belly and the excision of a segment of the tendon to avoid its restoration. PMID- 15109766 TI - Coming of age of lipid-based drug delivery systems. PMID- 15109767 TI - Tocol emulsions for drug solubilization and parenteral delivery. AB - Tocols represent a family of tocopherols, tocotrienols, and their derivatives, and are fundamentally derived from the simplest tocopherol, 6-hydroxy-2-methyl-2 phytylchroman, which is referred to as "tocol". The most common tocol is D-alpha tocopherol, also known as vitamin E. Tocols can be excellent solvents for water insoluble drugs and are compatible with other cosolvents, oils and surfactants. This review highlights the major developments in the use of tocols in parenteral emulsions for drug delivery, with a focus on drug solubilization, physicochemical properties, and biopharmaceutical applications. Tocol emulsions offer an appealing alternative for the parenteral administration of poorly soluble drugs, including major chemotherapeutics such as paclitaxel. Data will be presented on solubilization of paclitaxel, a key chemotherapeutic agent, and its corresponding formulation development, toxicity, efficacy and pharmacokinetic studies in animal models and humans. The breadth of the utility of tocol-based emulsions will be discussed and examples of specific therapeutic drugs and applications will be provided. As these formulations progress further in the clinic, the therapeutic utility of tocol emulsions is anticipated to expand. PMID- 15109768 TI - Solid lipid nanoparticles for parenteral drug delivery. AB - This review describes the use of nanoparticles based on solid lipids for the parenteral application of drugs. Firstly, different types of nanoparticles based on solid lipids such as "solid lipid nanoparticles" (SLN), "nanostructured lipid carriers" (NLC) and "lipid drug conjugate" (LDC) nanoparticles are introduced and structural differences are pointed out. Different production methods including the suitability for large scale production are described. Stability issues and drug incorporation mechanisms into the particles are discussed. In the second part, the biological activity of parenterally applied SLN and biopharmaceutical aspects such as pharmacokinetic profiles as well as toxicity aspects are reviewed. PMID- 15109769 TI - Micelles from lipid derivatives of water-soluble polymers as delivery systems for poorly soluble drugs. AB - Polymeric micelles have a whole set of unique characteristics, which make them very promising drug carriers, in particular, for poorly soluble drugs. Our review article focuses on micelles prepared from conjugates of water-soluble polymers, such as polyethylene glycol (PEG) or polyvinyl pyrrolidone (PVP), with phospholipids or long-chain fatty acids. The preparation of micelles from certain polymer-lipid conjugates and the loading of these micelles with various poorly soluble anticancer agents are discussed. The data on the characterization of micellar preparations in terms of their morphology, stability, longevity in circulation, and ability to spontaneously accumulate in experimental tumors via the enhanced permeability and retention (EPR) effect are presented. The review also considers the preparation of targeted immunomicelles with specific antibodies attached to their surface. Available in vivo results on the efficiency of anticancer drugs incorporated into plain micelles and immunomicelles in animal models are also discussed. PMID- 15109770 TI - Therapeutic applications of lipid-coated microbubbles. AB - Lipid-coated microbubbles represent a new class of agents with both diagnostic and therapeutic applications. Microbubbles have low density. Stabilization of microbubbles by lipid coatings creates low-density particles with unusual properties for diagnostic imaging and drug delivery. Perfluorocarbon (PFC) gases entrapped within lipid coatings make microbubbles that are sufficiently stable for circulation in the vasculature as blood pool agents. Microbubbles can be cavitated with ultrasound energy for site-specific local delivery of bioactive materials and for treatment of vascular thrombosis. The blood-brain barrier (BBB) can be reversibly opened without damaging the neurons using ultrasound applied across the intact skull to cavitate microbubbles within the cerebral microvasculature for delivery of both low and high molecular weight therapeutic compounds to the brain. The first lipid-coated PFC microbubble product is currently marketed for diagnostic ultrasound imaging. Clinical trials are currently in process for treatment of vascular thrombosis with ultrasound and lipid-coated PFC microbubbles (SonoLysis Therapy). Targeted microbubbles and acoustically active PFC nanoemulsions with specific ligands can be developed for detecting disease at the molecular level and targeted drug and gene delivery. Bioactive compounds can be incorporated into these carriers for site-specific delivery. Our aim is to cover the therapeutic applications of lipid-coated microbubbles and PFC emulsions in this review. PMID- 15109771 TI - Potential role of the low-density lipoprotein receptor family as mediators of cellular drug uptake. AB - We highlight the importance of the low-density lipoprotein (LDL) receptor family and its pharmaceutical implications in the field of drug delivery. The members of the LDL receptor family are a group of cell surface receptors that transport a number of macromolecules into cells through a process called receptor-mediated endocytosis. This process involves the receptor recognizing a ligand from the extracellular membrane (ECM), internalizing it through clathrin-coated pits and degrading it upon fusion with lysosomes. There are nine members of the receptor family, which include the LDL receptor, low-density lipoprotein-related protein (LRP), megalin, very low-density lipoprotein (VLDL) receptor, apoER2 and sorLA/LRP11, LRP1b, MEGF7, LRP5/6; the former six having been identified in humans. Each member is expressed in a number of different tissues and has a wide range of different ligands, not specific to the recognition of the LDL particle. Thus, rather than the original hypothesis that the receptor is only a mediator of cholesterol uptake, it may also be involved in a number of other physiological functions, including the progression of certain disease states and, potentially, cellular drug uptake. A number of studies have suggested that the LDL receptors are involved in endocytosis of drugs and drug formulations including aminoglycosides, anionic liposomes and cyclosporine A (CsA). This article reviews the importance of lipoproteins as a drug delivery system and how LDL receptors are relevant to the design and targeting of specific drugs. PMID- 15109772 TI - Pushing the envelope on molecular dating. PMID- 15109773 TI - Regulating gene expression: surprises still in store. PMID- 15109774 TI - Mitochondrial DNA repeats constrain the life span of mammals. PMID- 15109775 TI - Evidence that functional transcription units cover at least half of the human genome. AB - Transcriptome analyses have revealed that a large proportion of the human genome is transcribed. However, many of these transcripts might be functionless. To distinguish functional transcription units (FTUs) from spurious transcripts, we searched for the hallmarks of selective pressure against mutations that impair transcription. We analyzed the distribution of transposable elements, which are counter selected within FTUs. We show that these features are sufficiently informative to predict whether a sequence is transcribed and, if transcribed, in which orientation. Our results indicate that FTUs constitute at least 50% of the genome and that approximately one-third of these transcripts apparently do not encode proteins. PMID- 15109776 TI - The impact of very short alternative splicing on protein structures and functions in the human genome. PMID- 15109777 TI - Molecular timescales and the fossil record: a paleontological perspective. PMID- 15109778 TI - Precision of molecular time estimates. PMID- 15109779 TI - Compactness of human housekeeping genes: selection for economy or genomic design? PMID- 15109780 TI - Genome mosaicism and organismal lineages. PMID- 15109781 TI - Dinoflagellate chloroplasts--where have all the genes gone? PMID- 15109782 TI - Phenotypic neighborhood and micro-evolvability. PMID- 15109783 TI - Ecology of prokaryotic viruses. AB - The finding that total viral abundance is higher than total prokaryotic abundance and that a significant fraction of the prokaryotic community is infected with phages in aquatic systems has stimulated research on the ecology of prokaryotic viruses and their role in ecosystems. This review treats the ecology of prokaryotic viruses ('phages') in marine, freshwater and soil systems from a 'virus point of view'. The abundance of viruses varies strongly in different environments and is related to bacterial abundance or activity suggesting that the majority of the viruses found in the environment are typically phages. Data on phage diversity are sparse but indicate that phages are extremely diverse in natural systems. Lytic phages are predators of prokaryotes, whereas lysogenic and chronic infections represent a parasitic interaction. Some forms of lysogeny might be described best as mutualism. The little existing ecological data on phage populations indicate a large variety of environmental niches and survival strategies. The host cell is the main resource for phages and the resource quality, i.e., the metabolic state of the host cell, is a critical factor in all steps of the phage life cycle. Virus-induced mortality of prokaryotes varies strongly on a temporal and spatial scale and shows that phages can be important predators of bacterioplankton. This mortality and the release of cell lysis products into the environment can strongly influence microbial food web processes and biogeochemical cycles. Phages can also affect host diversity, e.g., by 'killing the winner' and keeping in check competitively dominant species or populations. Moreover, they mediate gene transfer between prokaryotes, but this remains largely unknown in the environment. Genomics or proteomics are providing us now with powerful tools in phage ecology, but final testing will have to be performed in the environment. PMID- 15109784 TI - Regulation of virulence determinants in Staphylococcus aureus: complexity and applications. AB - The virulence of Staphylococcus aureus is essentially determined by cell wall associated proteins and secreted toxins that are regulated and expressed according to growth phases and/or growth conditions. Gene expression is regulated by specific and sensitive mechanisms, most of which act at the transcriptional level. Regulatory factors constitute numerous complex networks, driving specific interactions with target gene promoters. These factors are largely regulated by two-component regulatory systems, such as the agr, saeRS, srrAB, arlSR and lytRS systems. These systems are sensitive to environmental signals and consist of a sensor histidine kinase and a response regulator protein. DNA-binding proteins, such as SarA and the recently identified SarA homologues (SarR, Rot, SarS, SarT, SarU), also regulate virulence factor expression. These homologues might be intermediates in the regulatory networks. The multiple pathways generated by these factors allow the bacterium to adapt to environmental conditions rapidly and specifically, and to develop infection. Precise knowledge of these regulatory mechanisms and how they control virulence factor expression would open up new perspectives for antimicrobial chemotherapy using key inhibitors of these systems. PMID- 15109785 TI - Genetic aspects of targeted insertion mutagenesis in yeasts. AB - Targeted insertion mutagenesis is a main molecular tool of yeast science initially applied in Saccharomyces cerevisiae. The method was extended to fission yeast Schizosaccharomyces pombe and to "non-conventional" yeast species, which show specific properties of special interest to both basic and applied research. Consequently, the behaviour of such non-Saccharomyces yeasts is reviewed against the background of the knowledge of targeted insertion mutagenesis in S. cerevisiae. Data of homologous integration efficiencies obtained with circular, ends-in or ends-out vectors in several yeasts are compared. We follow details of targeted insertion mutagenesis in order to recognize possible rate-limiting steps. The route of the vector to the target and possible mechanisms of its integration into chromosomal genes are considered. Specific features of some yeast species are discussed. In addition, similar approaches based on homologous recombination that have been established for the mitochondrial genome of S. cerevisiae are described. PMID- 15109786 TI - Comparative cell wall core biosynthesis in the mycolated pathogens, Mycobacterium tuberculosis and Corynebacterium diphtheriae. AB - The recent determination of the complete genome sequence of Corynebacterium diphtheriae, the aetiological agent of diphtheria, has allowed a detailed comparison of its physiology with that of its closest sequenced pathogenic relative Mycobacterium tuberculosis. Of major importance to the pathogenicity and resilience of the latter is its particularly complex cell envelope. The corynebacteria share many of the features of this extraordinary structure although to a lesser level of complexity. The cell envelope of M. tuberculosis has provided the molecular targets for several of the major anti-tubercular drugs. Given a backdrop of emerging multi-drug resistant strains of the organism (MDR-TB) and its continuing global threat to human health, the search for novel anti-tubercular agents is of paramount importance. The unique structure of this cell wall and the importance of its integrity to the viability of the organism suggest that the search for novel drug targets within the array of enzymes responsible for its construction may prove fruitful. Although the application of modern bioinformatics techniques to the 'mining' of the M. tuberculosis genome has already increased our knowledge of the biosynthesis and assembly of the mycobacterial cell wall, several issues remain uncertain. Further analysis by comparison with its relatives may bring clarity and aid the early identification of novel cellular targets for new anti-tuberculosis drugs. In order to facilitate this aim, this review intends to illustrate the broad similarities and highlight the structural differences between the two bacterial envelopes and discuss the genetics of their biosynthesis. PMID- 15109787 TI - Studying the dynamics of microbial populations during food fermentation. AB - The dynamics of growth, survival and biochemical activity of microorganisms in food are the result of stress reactions in response to the changing of the physical and chemical conditions into the food microenvironment, the ability to colonise the food matrix and to growth into a spatial heterogeneity, and the in situ cell-to-cell ecological interactions which often happen in a solid phase. In food, ecological approaches to study the evolution of microbial flora would be useful to comprehend better the microbiological processes involved in food processing and ripening, to improve microbiological safety by monitoring in situ pathogenic bacteria, and to evaluate the effective compositions of the microbial populations. This paper gives a general overview of biotechnological approaches to study microbial populations in food fermentation. PMID- 15109788 TI - The effect of abrupt osmotic shifts on the lag phase duration of physiologically distinct populations of Salmonella typhimurium. AB - Relative lag time (RLT), i.e. lag time divided by generation time, was used to characterise the lag phase response of exponential and stationary phase Salmonella typhimurium subjected to NaCl-mediated hyperosmotic shifts. Abrupt hyperosmotic shifts induced lag phases. The RLT, however, varied with the physiological history of the inoculum and the magnitude of the shift. Turbidimetric data showed that exponential phase cells had larger RLTs (up to approximately 8 units) than stationary phase cells (up to 2-4 units). Inocula containing exponential and stationary phase cells mixed in known proportions gave intermediate results. For viable count data, there was little difference in RLT between exponential and stationary phase cells. The RLT response determined turbidimetrically was reproducible for exponential phase cells, but less so for stationary phase cells. It is suggested that there may be a lower limit for resolution of RLT, in the range 0-2 units, and that this may account for the lack of reproducibility in RLTs of stationary phase cells. It is hypothesised that stationary phase cells have enhanced resistance to osmotic stress and are able to exploit new growth environments at low a(w) more rapidly than exponential phase cells, resulting in shorter lag phases. However, the data indicate that turbidimetry may not accurately describe the lag phase response of exponential phase cells subjected to large osmotic shifts. Viable count data is required to investigate this hypothesis further. PMID- 15109789 TI - The use of chlorine dioxide to control Alicyclobacillus acidoterrestris spores in aqueous suspension and on apples. AB - Alicyclobacillus acidoterrestris, a thermoacidophilic, spore-forming bacterium, has been identified as a spoilage organism in commercial, pasteurized fruit juices. This study was undertaken to evaluate chlorine dioxide for reducing numbers of A. acidoterrestris spores on laboratory media and on apples. A. acidoterrestris spores in aqueous suspension and on apple surfaces of four different cultivars were treated with several concentrations of chlorine dioxide. Spores in aqueous suspension treated with 40 ppm for 5 min were reduced by more than 4 log. Treatment with 80 ppm for 1 min and 120 ppm for 30 s resulted in about 1.8 log and 4.8 log reductions of spore viability, respectively, and treatment at 80 and 120 ppm for 5 min reduced spore viability to undetectable levels (<0.7 log CFU/ml). When applied to the surfaces of four different apple cultivars ('Red Delicious', 'Golden Delicious', 'Gala', and 'Fuji'), 40 ppm free chlorine dioxide reduced A. acidoterrestris spore numbers by 1.5, 3.2, 4.5, >4.8 log after 1-, 2-, 3-, and 4-min treatments, respectively. Spore numbers were reduced by >4.8 log with 120 ppm free chlorine dioxide after only 1-min treatment. However, there was no significant difference between apple cultivars (P>0.05) on spore reduction. These results show the great effectiveness of chlorine dioxide in controlling A. acidoterrestris spores both in aqueous suspension and on apple surfaces. There was no synergistic effect on spore reduction when chlorine dioxide treatment of aqueous suspension was followed by heat. PMID- 15109790 TI - The antilisterial effect of Leuconostoc carnosum 4010 and leucocins 4010 in the presence of sodium chloride and sodium nitrite examined in a structured gelatin system. AB - To further enhance biopreservation of meat products, the antilisterial effect of the newly described protective culture Leuconostoc carnosum 4010 and its bacteriocins, leucocins 4010, was examined in the presence of sodium chloride and sodium nitrite in a solid matrix using a structured gelatin system. Interaction between Listeria monocytogenes 4140 and Leuc. carnosum 4010 or the leucocins 4010 resistant mutant L. monocytogenes 4140P showed that the inhibitory effect of Leuc. carnosum 4010 in the gelatin system was caused by the production and activity of leucocins 4010. The presence of sodium chloride (2.5% w/v) and sodium nitrite (60 mg/l) reduced the antilisterial effect of Leuc. carnosum 4010 in the structured gel system compared to the use of Leuc. carnosum 4010 alone. Investigations carried out at 10 degrees C showed that the lag phase of L. monocytogenes 4140 in the presence of Leuc. carnosum 4010 was reduced from 71 to 58 h by the addition of sodium chloride and to 40 h by the addition of sodium nitrite. Addition of sodium chloride increased the maximum specific growth rate of L. monocytogenes 4140 in the presence of Leuc. carnosum 4010 from 0.02 to 0.06 h(-1), whereas no change was observed by the addition of sodium nitrite. Compared to the antilisterial effect of leucocins 4010 alone, the addition of sodium chloride (2.5%, w/v) decreased the antilisterial effect at high concentrations of leucocins 4010 (5.3 and 10.6 AU/ml) as measured after 11 days of incubation at 10 degrees C. In gels with added leucocins 4010, the most pronounced reduction in growth of L. monocytogenes 4140 was observed at the highest concentration of leucocins 4010 (10.6 AU/ml) together with sodium nitrite (60 mg/l). More detailed information on the lag phase and the maximum specific growth rate of single colonies of L. monocytogenes 4140 in the presence of leucocins 4010 was obtained using microscopy and image analysis. No pronounced difference in the growth of single colonies was observed in the gel system. Real-time measurements of colony growth at 10 degrees C in the gelatin matrix showed that the growth inhibiting effect of leucocins 4010, including a longer lag phase as well as a lower maximum specific growth rate for L. monocytogenes 4010, was negated in the presence of 2.5% (w/v) sodium chloride. PMID- 15109791 TI - Indigenous raw milk microbiota influences the bacterial development in traditional cheese from an alpine natural park. AB - Nostrano di Primiero is a 6-month ripened cheese produced from raw milk collected in the Paneveggio-Pale di San Martino Natural Park area in the Italian Dolomites. In summer, this cheese is made using milk collected from two different areas, Passo Rolle and Vanoi, in the Paneveggio Natural Park. During the experiment, the milk from the two areas was separately processed, and cheeses were made in the same cheese factory using the same technological process. The microbiota of raw milk and cheeses of the two areas was isolated and the dominant population was monitored by RAPD analysis and identified by 16S rRNA sequence. The milk of the Passo Rolle area was mainly composed of mesophilic strains, thermophilic Streptococcus thermophilus, and low amounts of enterococci were also found; the milk of the Vanoi area was dominated by mesophilic microbiota mostly Lactococcus lactis ssp. cremoris and ssp. lactis and Lactobacillus paracasei ssp. paracasei. The plating of the natural starter culture revealed the presence of a relevant community of thermophilic cocci and lower amounts of enterococci. The dynamic population analysis showed the importance of the natural starter culture in the first 2 days of cheese ripening in both cheeses. Moreover, the large biodiversity observed in the raw milks was also detected in the cheeses during ripening. The Vanoi cheese was dominated by Enterococcus faecium and Streptococcus macedonicus in the first two days and mesophilic 21 Lb. paracasei ssp. paracasei became the most represented population after 15 days of ripening. In the first few days, the Rolle cheese was characterized by being mainly composed of thermophilic S. macedonicus and S. thermophilus and secondarily by mesophilic cocci. During ripening, the microbiota composition changed, and at 15 days, mesophilic lactobacilli were the dominant population, but later, this was mainly composed of mesophilic cocci and lactobacilli. The taxonomical identification by 16S rRNA sequence confirmed a large biodiversity related to raw milk microbiota and only five strains of S. macedonicus, Lactobacillus plantarum, 21 Lb. paracasei ssp. paracasei, Lactobacillus fermentum and E. faecium were detected in both cheeses. PMID- 15109792 TI - Characterization of thoeniicin 447, a bacteriocin isolated from Propionibacterium thoenii strain 447. AB - Fifteen strains of propionibacteria, isolated from dairy products, were screened for the production of bacteriocins. Propionibacterium thoenii 447 produced an antimicrobial peptide, thoeniicin 447, which acted bactericidal against Lactobacillus delbrueckii subsp. bulgaricus and bacteriostatic against Propionibacterium acnes. Thoeniicin 447 remained active after 15 min at 100 degrees C and after 30 min of incubation at pH 1-10. The peptide was inactivated when treated with pepsin, pronase, alpha-chymotrypsin, trypsin and proteinase K. Optimal bacteriocin production was detected during late exponential growth. The peptide was partially purified by ammonium sulfate precipitation, followed by SP Sepharose cation exchange chromatography. The estimated size of thoeniicin 447, according to tricine-SDS-PAGE, is 6 kDa. Based on DNA sequencing, the mature peptide is 7130.20 Da in size and homologous to propionicin T1, produced by P. thoenii strain 419 (=NCFB 568(T)). Strain 447 is phenotypically different from strain 419 and belongs to a separate ribotype cluster. To our knowledge, this is the first report of a bacteriocin from a Propionibacterium species active against P. acnes. PMID- 15109793 TI - Pulsed field gel electrophoresis of related Escherichia coli O157 isolates associated with beef cattle and comparison with unrelated isolates from animals, meats and humans. AB - The pulsed field gel electrophoresis (PFGE) diversity of 51 related Escherichia coli O157 isolates, associated with beef cattle from a single-farm-to-single abattoir (SF-SA) chain of events was determined. The 51 related E. coli O157 isolates from hides, faeces or carcasses of SF-SA cattle produced 11 different PFGE profiles. Also, the PFGE diversity of 6 isolates, associated with a second cattle abattoir, was determined; only two PFGE profiles were found. On the other hand, the PFGE diversity of 136 unrelated E. coli O157 isolates (from healthy meat animals, retail meats and cases of human disease) was also determined. The 136 unrelated E. coli O157 isolates produced 78 different PFGE profiles, most of which (approximately 70%) comprised only one isolate. Overall, the results showed: (a) related E. coli O157 isolates (from both SF-SA events, and the second abattoir) had a markedly narrower clonal profile than the 136 unrelated E. coli O157 isolates; (b) the isolation of identical PFGE types from hide, lairage environment, and carcasses confirms the significance of cross-contamination (both pre-slaughter and during skinning) taking place at abattoirs; and (c) PFGE typing of isolates offers a good tool for tracking sources/routes of such cross contamination. Such cross-contamination may lead to originally E. coli O157-free animals (and resultant carcasses) becoming contaminated during farm-slaughter dressing chain of events, so development of efficient control strategies is required. PMID- 15109794 TI - Dormancy, activation and viability of Rhizopus oligosporus sporangiospores. AB - Interruption of dormancy to improve viability of Rhizopus oligosporus sporangiospores is crucial for the application of stored starter cultures for fungal (tempe) production. We aimed to assess the extent of dormancy and factors that could result in activation. Whereas heat treatments were unsuccessful, Malt Extract Broth (MEB) showed to be a good activation medium, with 80% of dormant spores being activated as measured by fluorescence microscopy using a fluorescent marker, compared with 11% with the control. Peptone and yeast extract but not glucose played an important role in activating dormant spores. Metabolically active (fluorescent) and swollen spores, followed by germ tubes were obtained after activation in MEB for 25 min., 2 and 4 h, respectively, at 37 degrees C. Simultaneously, some interesting transitions took place. Dormant spores represent 85-90% of the total spores at harvest and after drying. Their number decreased to 21-32% after activation with MEB with a concomitant increase of metabolically active spores. As a result of storage, some dormancy was lost, yielding an increase of active spores from 11.2% at harvest to 28.8% after 3 months storage. Levels of active spores were well correlated with their viability. By activation of dormant spores, their viability increased; levels of viable and active spores were maximum in 1 month old starter (61.7% and 75.9% of total spores, respectively) but gradually decreased with concomitant increase of the number of dead spores. PMID- 15109795 TI - Antimicrobial and antioxidative enrichment of oak (Quercus robur) bark by rotation planar extraction using ExtraChrom. AB - The multifunctional ExtraChrom instrument was used in the extraction of antimicrobial and radical scavenging components from oak (Quercus robur L.) bark. Milled and sieved oak bark was extracted with 80% (v/v) methanol solution in water on the ExtraChrom instrument using step-gradient in the preparative separation. Extracts were tested using agar diffusion method on Staphylococcus aureus, Enterobacter aerogenes and Candida albicans. Some extracts showed moderate bactericidal, fungicidal, bacteriostatic and fungistatic activity. The composition related to activity of the fractions and extracts was screened simultaneously by thin-layer chromatography (TLC) detected by UV and by spraying the plate with radical scavenging reagent 1,1-diphenyl-2-picrylhydrazyl (DPPH) to detect antioxidant activity. Thus, we could demonstrate the antiradical and antimicrobial activity of oak beneficial in the storage of wine against the oxidation and human microbial exposure. PMID- 15109796 TI - Use of novel PCR primers specific to the genes of staphylococcal enterotoxin G, H, I for the survey of Staphylococcus aureus strains isolated from food-poisoning cases and food samples in Taiwan. AB - Data regarding the incidence of the newly found enterotoxigenic Staphylococcus aureus strains in food poisoning cases and in food samples were to date not available in Taiwan. In this study, PCR primers specific for the detection of SEG, H and I genes, i.e., seg, seh and sei, were used for the assay of 55 human isolates of S. aureus negative to the classical enterotoxins (SEA-->SEE) detection. These isolates were from the fecal specimens of the patients suffering from food poisoning outbreaks. Only eight strains were found to have the seg, seh and sei. The presence of other bacterial pathogens, such as Vibrio parahaemolyticus, Bacillus cereus and Salmonella spp. and perhaps, strains producing other new staphylococcal enterotoxins, in the fecal specimens of these patients, may account for these food poisoning cases. For 139 strains from food samples, such as frozen Chinese foods, Chinese sausages and lunch meals, sea strains accounted for the major portion and it seemed to be the most common SE type to coexist with seg, seh and sei. Only two strains had sec and none of them had seg, seh or sei. For strains without the classical SE genes, only 13 strains had seg, seh and/or sei. The above results imply that seg, seh and sei S. aureus strains play only a minor role in food-borne outbreaks in Taiwan. PMID- 15109797 TI - Have changes to meat and poultry food safety regulation in Australia affected the prevalence of Salmonella or of salmonellosis? AB - During the 1990s, there was radical change in regulation of meat and poultry hygiene in Australia, and Australian Standards were developed for each sector of the meat industry. Systems for industry/government co-regulation and company employed meat inspection were introduced based on company HACCP programs approved and audited by the Controlling Authority. However, in the 5 years since regulatory changes took full effect, rates of salmonellosis have not decreased (surveillance and reporting systems have remained unchanged). Using statistics gathered by the National Enteric Pathogens Surveillance Scheme, an attempt was made to link Salmonella serovars isolated from meat and poultry with those causing salmonellosis. Two periods were studied, 1993/1994, before regulations were introduced, and 2000/2001, when regulations should be having an effect. For red meat, the same serovars were prominent among the top 10 isolates both before and after regulation, and there was little linkage with salmonelloses. For poultry, frequently isolated serovars differed pre- and post-regulation, however, in both periods there was some linkage between serovars isolated from poultry and those causing salmonelloses. Using published and unpublished survey data, it was concluded that there had been improvements in microbiological quality of red meat and poultry over the same timeframe as regulatory changes. That these improvements apparently have not carried through to reduced case-rates for salmonellosis may be due to numerous causes, including lack of control in the food processing, food service and home sectors. The present paper illustrates difficulties faced by governments in measuring public health outcomes of changes to food hygiene regulation. PMID- 15109798 TI - Susceptibility of Vibrio parahaemolyticus to various environmental stresses after cold shock treatment. AB - Vibrio parahaemolyticus was subjected to cold shock treatment at 20 or 15 degrees C for 2 or 4 h. The effect of cold shock on the survival of V. parahaemolyticus subjected to subsequent low temperature (5 and -18 degrees C) and other adverse conditions (47 degrees C, 6 ppm crystal violet, 1000 ppm H(2)O(2), 25 mM acetic acid and 25 mM lactic acid) was investigated. Regardless of the cold shock treatment, survival of V. parahaemolyticus increased when stored at 5 or -18 degrees C, while no increase in survival was noted for cells cold shocked in the presence of chloramphenicol. Cold shock treatment under the conditions tested, in general, enabled V. parahaemolyticus cells to survive better following subsequent challenge by crystal violet, while the cold-shocked organism was more susceptible to high temperature (47 degrees C), H(2)O(2) and organic acids (lactic and acetic acid) than the non-shocked cells. Furthermore, the temperature and time of the cold shock treatment affected the cold shock response of V. parahaemolyticus. PMID- 15109799 TI - Microbial contamination on beef in relation to hygiene assessment based on criteria used in EU Decision 2001/471/EC. AB - Thirty-six carcasses were sampled over a 12-month period at an Irish beef abattoir. Between one and five carcass sites (including the hock, brisket, cranial back, bung, inside round and outside round) were sampled after hind leg skinning, hide removal, bung tying, evisceration, splitting, washing, chilling for 24 h and boning, using a wet and dry, cotton wool swab technique. For each sample, total viable counts (TVC), Escherichia coli, total coliforms and Enterobacteriaceae were enumerated. The results are considered in relation to European Union Decision 2001/471/EC which sets performance criteria for TVCs and Enterobacteriaceae in samples taken by excision. Though not explicitly stated in the Decision, it has been proposed that microbiological performance criteria for samples taken by swabbing be set at 20% of the values set for excision samples. Therefore, log mean TVCs in carcass swab samples taken before chilling are acceptable, marginal and unacceptable when they are <2.8, 2.8-4.30 and >4.30 cm( 2), respectively. By these criteria, TVCs on carcasses in the present study were in the marginal range. The marginal result for TVCs was due in the most part to hide removal operations, particularly at the hock and brisket sites. Bacterial contamination on post-chill carcasses was similar or lower to that on pre-chill carcasses, while boning resulted in general increases in TVCs and in E. coli, total coliform and Enterobacteriaceae numbers. In Decision 2001/471/EC, the effects of chilling and boning are not included in the assessment of process control. Data from this study indicate that performance criteria based on log mean Enterobacteriaceae values are unsuitable because of the infrequent occurrence of these organisms on the carcass. PMID- 15109800 TI - Inactivation of Bacillus cereus spores in milk by mild pressure and heat treatments. AB - The objective of this work was to study the germination and subsequent inactivation of Bacillus cereus spores in milk by mild hydrostatic pressure treatment. In an introductory experiment with strain LMG6910 treated at 40 degrees C for 30 min at 0, 100, 300 and 600 MPa, germination levels were 1.5 to 3 logs higher in milk than in 100 mM potassium phosphate buffer (pH 6.7). The effects of pressure and germination-inducing components present in the milk on spore germination were synergistic. More detailed experiments were conducted in milk at a range of pressures between 100 and 600 MPa at temperatures between 30 and 60 degrees C to identify treatments that allow a 6 log inactivation of B. cereus spores. The mildest treatment resulting in a 6 log germination was 30 min at 200 MPa/40 degrees C. Lower treatment pressures or temperatures resulted in considerably less germination, and higher pressures and temperatures further increased germination, but a small fraction of spores always remained ungerminated. Further, not all germinated spores were inactivated by the pressure treatment, even under the most severe conditions (600 MPa/60 degrees C). Two possible approaches to achieve a 6 log spore inactivation were identified, and validated in three additional B. cereus strains. The first is a single step treatment at 500 MPa/60 degrees C for 30 min, the second is a two-step treatment consisting of pressure treatment for 30 min at 200 MPa/45 degrees C to induce spore germination, followed by mild heat treatment at 60 degrees C for 10 min to kill the germinated spores. Reduction of the pressurization time to 15 min still allows a 5 log inactivation. These results illustrate the potential of high pressure treatment to inactivate bacterial spores in minimally processed foods. PMID- 15109801 TI - Exposing relationships using directed evolution. PMID- 15109802 TI - Nuclear remodeling after SCNT: a contractor's nightmare. PMID- 15109803 TI - Realization of beta-lactamase as a versatile fluorogenic reporter. PMID- 15109804 TI - A reason to be optimistic about biodiesel: seed meal as a valuable soil amendment. PMID- 15109805 TI - The future of patent deposition of microorganisms? PMID- 15109806 TI - Biotechnology, bioethics and anti-aging interventions. PMID- 15109807 TI - Food forensics: using DNA technology to combat misdescription and fraud. PMID- 15109808 TI - Using electropolymerized non-conducting polymers to develop enzyme amperometric biosensors. PMID- 15109809 TI - Plant bioinformatics: from genome to phenome. PMID- 15109810 TI - A revival of bispecific antibodies. PMID- 15109811 TI - Metabolomics by numbers: acquiring and understanding global metabolite data. PMID- 15109812 TI - Recent developments in the optimization of thermostable DNA polymerases for efficient applications. PMID- 15109813 TI - RT-PCR detection of yellow head virus (YHV) infection in Penaeus monodon using dried haemolymph spots. AB - Sample collection and RNA isolation from shrimp haemolymph for RT-PCR diagnosis of yellow head virus (YHV) infections is crucial for disease control programs for cultivated shrimp in Thailand. Problems with RNA degradation arise when field samples must be collected far from the laboratory by relatively inexperienced personnel who do not have ready access to sophisticated reagents. In an attempt to solve this problem, haemolymph samples from shrimp were collected either by mixing with 10% (w/v) sodium citrate or by spotting on ISOCODE filter paper. RNA was extracted subsequently either by a rapid boiling method or by using TRI reagent and the extracts were used in a semi-quantitative, non-stop, semi-nested RT-PCR assay for YHV. Dried haemolymph spots on ISOCODE filter paper extracted with TRI reagent gave the most reliable and reproducible results. It also allowed longer periods of storage at room temperature. PMID- 15109814 TI - Removal of hepatitis A virus from water by polyacrylonitrile-based ultrafiltration membranes. AB - Ultrafiltration-based polyacrylonitrile membranes (UF-membranes) were evaluated for their ability to retain tissue culture adapted indigenous hepatitis A virus from water. Amicon cell (dead-end ultrafiltration unit)-based experiment was set up and viral assay was carried out using rapid and sensitive reverse transcriptase polymerase chain reaction (RT-PCR). The RT-PCR results show retention of virus particles by the polyacrylonitrile membranes. A protocol for routine virological evaluation of membranes is described and the use of these membranes for water purification units and for virus concentration systems for field application is discussed. PMID- 15109815 TI - Detection of white spot syndrome virus from stomach tissue homogenate of the kuruma shrimp (Penaeus japonicus) by reverse passive latex agglutination. AB - A reversed passive latex agglutination (RPLA) assay was developed for detecting the white spot syndrome virus (WSSV), which was formally named as penaeid rod shaped DNA virus (PRDV) in Japan, from stomach tissue homogenate of the kuruma shrimp (Penaeus japonicus). Using high-density latex particles and specific polyclonal antibody, WSSV was detectable after 4h incubation. The hemolymph, the stomach, and the gills were extracted from a shrimp that had been infected experimentally with WSSV, the virus contained in each sample was tested by the PRLA and PCR assay. It was possible to detect the WSSV only from stomach tissue homogenates by the RPLA assay. And there was an agreement between RPLA and PCR assays for WSSV detection. Considering that the RPLA assay does not require biochemical expertise and latex reagents and all apparatus can be provided as a kit, this assay can be used for virus detection in the culture pond of shrimps or in the field as a convenient method. PMID- 15109816 TI - Development of Taqman RT-nested PCR system for clinical SARS-CoV detection. AB - Severe acute respiratory syndrome (SARS) is an acute newly emerged infectious respiratory illness. The etiologic agent of SARS was named 'SARS-associated coronavirus' (SARS-CoV) that can be detected with reverse transcription polymerase chain reaction (RT-PCR) assays. In this study, 12 sets of nested primers covering the SARS-CoV genome have been screened and showed sufficient sensitivity to detect SARS-CoV in RNA isolated from virus cultured in Vero 6 cells. To optimize further the reaction condition of those nested primers sets, seven sets of nested primers have been chosen to compare their reverse transcribed efficiency with specific and random primers, which is useful to combine RT with the first round of PCR into a one-step RT-PCR. Based on the sensitivity and simplicity of results, the no. 73 primer set was chosen as the candidate primer set for clinical diagnoses. To specify the amplicon to minimize false positive results, a Taqman RT-nested PCR system of no. 73 nested primer set was developed. Through investigations on a test panel of whole blood obtained from 30 SARS patients and 9 control persons, the specificity and sensitivity of the Taqman RT-nested PCR system was found to be 100 and 83%, respectively, which suggests that the method is a promising one to diagnose SARS in early stages. PMID- 15109817 TI - Immunofluorescence test for sensitive detection of varicella-zoster virus specific IgG: an alternative to fluorescent antibody to membrane antigen test. AB - A highly sensitive indirect fluorescence antibody test (IFAT) has been developed on the basis of varicella-zoster virus (VZV)-infected human lung carcinoma (A549) cells and evaluated for the determination of immunity to VZV. Different serum panels with negative, low, moderate or high anti-VZV IgG levels detected by the fluorescent antibody to membrane antigen (FAMA) assay were investigated. As a result, the sensitivity and the specificity of IFAT were 100% compared to FAMA test. In anti-VZV IgG-positive sera, a significant correlation between the results of FAMA procedure and IFAT could be shown. However, there were considerably higher antibody titers by the IFAT than by FAMA. Whereas the FAMA test had a detection limit of 250 mIU/ml anti-VZV IgG, the limit of detection of IFAT was 50 mIU/ml. In conclusion, the IFAT using VZV-infected A549 cells as antigen allows a highly sensitive, specific, and rapid detection of anti-VZV IgG class antibodies. This simple technique can replace the labor-intensive FAMA procedure for laboratory determination of immunity to VZV. PMID- 15109818 TI - Use of serial maternal urine cytomegalovirus PCR to detect primary CMV infection in seronegative pregnant women. AB - The aim of the study was to determine if serial maternal urine polymerase chain reaction (PCR) tests can detect primary CMV infection during pregnancy. This was a prospective study conducted from 1 January 1999 to 31 December 1999 in an antenatal clinic setting of a teaching hospital. The study group included women who were CMV IgG negative and aged <30 years or had a pre-school child. They were invited to self-collect urine samples monthly and send them to the laboratory by post. Cord bloods were tested for CMV IgG to detect seroconversion. An anxiety questionnaire was sent to all study participants. At first attendance, 1549 (42%) women were CMV IgG negative. Of the 696 eligible women, 609 (88%) participated in the urine PCR study. PCR was performed on 2263 urine samples (median of 4/pregnancy). Primary CMV infection was identified in one woman by urine PCR at 36 weeks (baby CMV negative). Cord blood samples were available from 152/609 infants (25%). Seroconversion was noted in only one woman. Replies to the questionnaire were received from 264/609 women (43%): 214 (81%) had little or no anxiety, and 220 (83%) felt reassured by their study participation. Serial urine PCR is a feasible method of detecting primary maternal CMV infection during pregnancy which has potential for evaluation in further studies. PMID- 15109819 TI - Early and rapid detection of enterovirus 71 infection by an IgM-capture ELISA. AB - Enterovirus 71 infection is more likely to induce severe complications and mortality than other enteroviruses. Laboratory diagnosis of enterovirus 71 in Taiwan still relies mainly on conventional virus isolation techniques that often require 5-10 days to obtain a result, thus hindering seriously the subsequent treatment and disease control measures. This study was to find a better alternative by developing a rapid enterovirus 71 detecting procedure, which can afford an earlier diagnosis and a more rapid outcome. In this study, an enterovirus 71-IgM-capture enzyme-linked immunosorbent assay (ELISA) was designed and tested with a total of 336 serum specimens collected from 236 cases of reported enterovirus infection with complications. Using virus isolation and neutralization test as standards, the sensitivity and specificity of the new protocol were 97.7 and 93.3%, respectively. Most of the IgM positive serum specimens were collected within 7 days after the onset of symptoms, while it appeared detectable up to 94 days after the onset of symptoms. Apart from being highly sensitive, rapid and low in cost, the new IgM-capture ELISA is sufficiently accurate to provide also reliable results for early detection of the virus. With this protocol, enterovirus 71 infections can be detected within 4h. PMID- 15109820 TI - Performance of ViroSeq HIV-1 Genotyping System in routine practice at a Belgian clinical laboratory. AB - Since there are indications of an increasing amount of non-B subtypes in Western Europe it was decided to assess the performance of the ViroSeq HIV-1 Genotyping System on a set of samples from the AIDS Reference Laboratory at the University Hospitals Leuven, a hospital with an increasing number of patients infected with non-B subtypes. The set consisted of 383 samples comprising 12 different subtypes and the genotyping kit was assessed for its amplification capabilities as well as its sequencing capabilities. Amplification failed in 32 samples (8.4%) and there was a tendency of a lower performance of the kit when it concerned the amplification of non-B subtypes. Regarding the sequencing performance of the HIV 1 Genotyping System, three different results could be considered. The performance of the entire set of primers (A, B, C, F, G and H) on the different subtypes showed a significant decrease of positive results for subtypes A, G and the recombinants whereas a tendency to less positive results could be detected for subtypes CRF12_BF, D, H and J. When looking at the performance of the individual primers for the different subtypes, only one result differed significantly: there were less positive results by applying primer F on subtype A. A tendency to less positive results was found for other combinations of primer and subtype, most of which comprised combinations with primers B, C, F and H. A final result was obtained by comparing the overall sequencing results of a certain primer on all the non-B subtypes with the results of the same primer on subtype B. Primer F showed significant less positive results and a tendency to less positive results was found for primer H. The other primers showed comparable results. All of the above results regarding the sequencing primers did not include primer D since this is a back-up primer for primer A. Analysis of the results for primer D showed that less positive results were found for all the non-B subtypes, most of which were significant. The overall performance of primer D on all non-B subtypes was only 15.7%. The use of primer D as a back-up primer was also investigated: it generated a positive result in only 17.3% of the cases where primer A failed. Most of these positive results were subtype B (74%). As a result of sequencing problems 65 out of 351 (18.5%) samples had to be processed with "in-house" procedures. PMID- 15109821 TI - A phage with high affinity for hepatitis B surface antigen for the detection of HBsAg. AB - An M13 phage, called PHH2, with the ability to bind HBsAg was isolated from a coat protein III display library. The region of the HBsAg polypeptide to which PHH2 binds was determined. The HBsAg binding phage was used in an assay referred to as "PHALISA", an abbreviation for phage-linked immune-absorbent assay. This assay was at least 20-100 times more sensitive in the detection of HBV antigen than conventional enzyme-linked immune-absorbent assays (ELISA). The application of this method for screening and detection of specific protein is discussed. PMID- 15109822 TI - Processing of plasmid DNA with ColE1-like replication origin. AB - With the increasing utilization of plasmid DNA as a biopharmaceutical drug, there is a rapidly growing need for high quality plasmid DNA for drug applications. Although there are several different kinds of replication origins, ColE1-like replication origin is the most extensively used origin in biotechnology. This review addresses problems in upstream and downstream processing of plasmid DNA with ColE1-like origin as drug applications. In upstream processing of plasmid DNA, regulation of replication of ColE1-like origin was discussed. In downstream processing of plasmid DNA, we analyzed simple, robust, and scalable methods, which can be used in the efficient production of pharmaceutical-grade plasmid DNA. PMID- 15109823 TI - Segregation of the yeast plasmid: similarities and contrasts with bacterial plasmid partitioning. AB - The high copy yeast plasmid 2 microm circle, like the well-studied low copy bacterial plasmids, utilizes two partitioning proteins and a cis-acting 'centromere'-like sequence for its stable propagation. Functionally, though, the protein and DNA constituents of the two partitioning systems are quite distinct. Key events in the yeast and bacterial segregation pathways are plasmid organization, localization, replication, 'counting' of replicated molecules and their distribution to daughter cells. We suggest that the two systems facilitate these common logistical steps by adapting to the physical, biochemical, and mechanical contexts in which the host chromosomes segregate. PMID- 15109824 TI - Isolation and sequencing of the replication region of plasmid pBFp1 isolated from a marine biofilm. AB - A 24 kb plasmid, pBFp1, encoding mercury resistance was previously isolated from a marine biofilm. Isolation and sequencing of a 4280 bp DNA fragment containing the plasmid replicon (rep-pBFp1) revealed a putative open reading frame encoding a RepA protein and an oriV-like region containing an A+T rich sub-region, iterons, and DnaA boxes. Sequence comparisons showed significant similarities to the incW plasmid pSa both for the RepA amino acid sequence and in the iteron DNA sequence. Plasmid pBFp1 was also shown to be incompatible with pSa in standard incompatibility testing. A probe from the repA gene of pBFp1 was further made and tested on a collection of plasmids exogenously isolated from marine habitats in a previous study. PMID- 15109825 TI - A gfp reporter plasmid to visualize Azorhizobium caulinodans during nodulation of Sesbania rostrata. AB - Compared with other labeling techniques, the use of the green fluorescent protein (GFP) is advantageous to visualize bacteria because observations can be performed in real time. This feature is particularly interesting to study invasion events of rhizobia during nodule development on their legume host plant. To investigate the symbiotic interaction between Azorhizobium caulinodans ORS571 and Sesbania rostrata, we constructed two plasmids, pMP220-hem-gfp5 and pBBR5-hem-gfp5-S65T, that carry a modified gfp gene, the expression of which is controlled by the constitutive hem promoter. Introduction of either of these plasmids into A. caulinodans allowed the visualization of single bacteria. Determination of the plasmid stability in cultured bacteria and in nodules demonstrated that pBBR5-hem gfp5-S65T is more stable than pMP220-hem-gfp5. The plasmid pBBR5-hem-gfp5-S65T can be used to study early invasion events during nodule development on hydroponic roots of S. rostrata. PMID- 15109826 TI - Complete sequence and structural organization of pFL5 and pFL7, two cryptic plasmids from Bacillus licheniformis. AB - The complete nucleotide sequences of two plasmids, pFL5 and pFL7, isolated from soil bacteria, Bacillus licheniformis FL5 and FL7, have been determined. The plasmids pFL5 and pFL7 were analyzed and found to be 9150 and 7853 bp in size with a G+C content of 41.0 and 43.6 mol%, respectively. Computer assisted analysis of sequence data revealed 11 possible ORFs in pFL5, four of which could be assigned no function from homology searches. Instead, eight putative ORFs were identified in pFL7, two of which appeared to have no biological function. All the ORFs were preceded by a ribosome binding site. The ORFs 9.5 and 6.7, each of 340 amino acids, were postulated to encode a replication protein similar to known replication proteins of rolling circle replicons, particularly those of the pC194 family. The structural organization of the two pFL plasmids is similar to the pTA plasmids family, with only a few putative coding regions that cannot be attributed to these plasmid backbone genes. In contrast to pTA plasmids, the majority of the genes have an orientation of transcription opposite to the direction of replication. The identified probable sso sequences seem to belong to a different group of those found in Bacillus plasmids; in fact, a significant level of homology was found with ssoA group sequences. These plasmids seem to be related to plasmids identified within the Bacillus subtilis group, confirming the low-level diversity among these replicons. PMID- 15109827 TI - Incompatibility and the partitioning site of the repABC basic replicon of the symbiotic plasmid from Rhizobium etli. AB - The basic replicon of the symbiotic plasmid (p42d) of Rhizobium etli CE3 is constituted by the repABC operon. Whereas RepC is essential for plasmid replication, RepA and RepB are involved in plasmid partitioning. Three incompatibility regions have been previously identified in this plasmid: the first one encodes RepA, a partitioning protein that also down-regulates the repABC transcription. The second region is situated within the repB-repC intergenic sequence (inc(alpha)), and the last one, inc(beta), is located in a 502 bp EcoRI fragment spanning the last 72-bp of the coding region of repC and the following downstream sequence. In this paper we show that: (1) The inc(beta) region is required for plasmid partitioning. (2) A 16-bp palindrome sequence, located 40 bp downstream of the repC gene of plasmid p42d, is necessary and sufficient to induce incompatibility towards the parental plasmid, and accounts for all the incompatibility properties of this region (inc(beta)). (3). The palindrome is the DNA target site for RepB binding. With these findings we propose that inc(beta) contains the partitioning site (par site) of the basic replicon of plasmid p42d, and that the 16-bp palindrome is the core sequence to nucleate the RepB binding. PMID- 15109828 TI - Construction of multi-purpose vectors, pCNS and pCNS-D2, are suitable for collection and functional study of large-scale cDNAs. AB - To efficiently perform collection and functional studies of large-scale cDNAs, we constructed multi-functional cDNA vectors for efficient full-length cDNA cloning, direct sequencing, easy screening, and the expression of cDNA in vitro and in vivo without subcloning the cDNA into other vectors. The constructed vectors, pCNS and pCNS-D2, contain a multi-cloning site for uni-directional full-length cDNA cloning, T7 and Sp6 RNA polymerase promoters for in vitro transcription and translation, and hCMV immediate early promoter and BGH poly(A) to allow expression in mammalian cells. Using these vectors, we constructed full-length enriched cDNA libraries containing 60-75% of the full-length cDNAs using two different oligo-capping methods. The subtracted cDNA libraries could also be constructed by removing of EF1-alpha cDNA, a highly expressed cDNA. In addition, we confirmed the translation of EF1-alpha cDNA in vitro and the expression of luciferase cDNA in mammalian cells. The expression efficiency of luciferase cDNA in different cell lines, such as HeLa, Hep3B, SNU638, and SNU668, showed that pCNS vectors can highly express target genes in different cell types. These results indicated that our multi-purpose vectors, pCNS and pCNS-D2, are useful tools for the construction of full-length cDNA libraries and high-throughput based functional study of cDNAs. PMID- 15109829 TI - Nucleotide sequence of the cryptic plasmid pTT8 from Thermus thermophilus HB8 and isolation and characterization of its high-copy-number mutant. AB - The complete nucleotide sequence of pTT8, a cryptic plasmid from Thermus thermophilus HB8, was determined. pTT8 was 9328bp long and its G+C content was 69%. pTT8 contained eight putative open reading frames, three of which showed extensive similarities to the plasmid addiction proteins PasA and PasB of pTC-F14 and pAM10.6, and the RepA protein of the ColE2-related plasmids, respectively. During the analysis of pTT8-based plasmid pPP442, which had been obtained during a promoter-screening experiment, we occasionally isolated a plasmid with a relatively high-copy-number. This plasmid, pPP442m, contained a 1025 bp fragment derived from the genome of the HB27 host strain immediately upstream of the putative repA gene. Using the ori region of pPP442m, we constructed an expression vector, pTEV131m, with an estimated high-copy-number of 30-40. This plasmid was stably maintained in T. thermophilus HB27 under nonselective conditions for at least 100 generations. Cloning of the alpha-amylase gene of Bacillus stearothermophilus DY-5 into pTEV131m gave more than twofold production of the enzyme compared with pTEV131, the parental plasmid. PMID- 15109830 TI - New shuttle vectors for ectopic insertion of genes into Bacillus subtilis. AB - We have constructed shuttle vectors for integration of genes via double homologous recombination into three ectopic sites on the chromosome of Bacillus subtilis. The sites of integration are the pyrD, gltA, and sacA genes located at 139 degrees, 172 degrees, and 333 degrees, respectively, on the chromosome. Integration of the vectors into the target genes leads to antibiotic resistance as well as different metabolic phenotypes. B. subtilis strains with integrations of the empty vectors were able to sporulate at rates comparable to wild type cells. Similar levels of expression were obtained from constitutive lacZ fusions integrated at the different sites. PMID- 15109831 TI - Improvement of pCVD442, a suicide plasmid for gene allele exchange in bacteria. AB - Allelic exchange experiments allow investigation of the functions of many unknown genes identified during the sequencing of entire genomes. Isogenic strains differing by only specific mutations can be constructed. Among other tools, suicide plasmids are widely used for this task. They present many advantages because they leave no scars on the chromosome, and therefore allow combining several mutations in the same genetic background. While using the previously described pCVD442 suicide plasmid [Infect. Immun. 59 (1991) 4310], we found untargeted recombination events due to the presence of an IS1 element on this plasmid. The plasmid was therefore improved by removal of the IS1 element. We also replaced the bla gene of pCVD442, conferring ampicillin resistance, by the cat gene conferring chloramphenicol resistance, leading to the new suicide plasmid pDS132. The plasmid was entirely sequenced. We demonstrate that this new vector can be easily used to introduce various types of mutations into different genetics backgrounds: removal of IS elements, introduction of point mutations or deletions. It can be introduced into bacterial strains by either transformation or conjugation. PMID- 15109832 TI - Development of an inducible system to control and easily monitor gene expression in Lactococcus lactis. AB - This report describes the implementation and use of a maltose-inducible system for regulated gene expression in Lactococcus lactis. The system was established using Green Fluorescent Protein as reporter. The transcription of a gene of interest from the inducible promoter of pLS1RGFP plasmid vector can be easily monitored by fluorescence spectroscopy and microscopy. As an example, the lactococcal ribonuclease III was overproduced in an active form. PMID- 15109833 TI - General and specialized vectors derived from pBM02, a new rolling circle replicating plasmid of Lactococcus lactis. AB - This paper reports the construction of several general cloning vectors and a specialized depurative vector based on a new lactococcal plasmid that replicates by the rolling circle mechanism [pBM02; Plasmid 49 (2003) 118]. Most vectors are shuttle vectors for Escherichia coli-Lactococcus lactis and carry replicons of both ColE1 and pBM02 plasmids (ColE1 is used even though the pBM02 replicon is fully active in both Gram-positive and Gram-negative organisms). Segregational and structural studies indicated that the new vectors were stable enough for the majority of applications. Further, since the basic replicon is compatible with plasmid derivatives of pWV01 and pSH71, they can be maintained in the same cell with members of the two largest vector series for L. lactis and other lactic acid bacteria, the pGK, and the pNZ series. PMID- 15109834 TI - Covalently immobilized thrombomodulin inhibits coagulation and complement activation of artificial surfaces in vitro. AB - Thrombomodulin (TM) serves as the endothelial cell receptor for thrombin and alters its characteristics from pro- to anticoagulant. Additionally, it promotes the formation of activated protein C. We evaluated the conservation of the overall outcome of these functions in recombinant TM linked to artificial surfaces by incubation with human whole blood in vitro. TM was covalently immobilized through poly(ethylene glycol) (PEG) spacers onto thin films of poly(octadecene alt maleic anhydride) covering planar glass substrates. TM binding to the polymer films was achieved after active ester formation at the carboxylic acid terminus of the PEG spacers and thoroughly characterized by HPLC based amino acid analysis, immunofluorescence and ellipsometry. TM-coated samples were incubated for 3h with freshly drawn whole human blood anticoagulated with heparin (5IU/ml) using in-house developed incubation systems. The substantially reduced activation of blood coagulation (TAT) for TM-coated samples correlates well with the degree of contact activation (bradykinin and FXIIa formation) while no significant effects were observed for the platelet activation (PF4). Further, complement activation (C5a levels), was strongly diminished at the TM-containing surfaces. We conclude that the suggested method for preparation of TM immobilization may serve to prepare model substrates for studies on TM interactions but similarly provides a promising coating strategy for blood contacting medical devices. PMID- 15109835 TI - Towards a fully-synthetic substitute of alginate: development of a new process using thermal gelation and chemical cross-linking. AB - We have previously described a gelation process based on the occurrence of both physical and a chemical mechanisms ('tandem process'), in which a telechelic linear poly(propylene glycol)-bl-poly(ethylene glycol)-bl-poly(propylene glycol) is first thermally gelled and subsequently covalently cross-linked by the reaction of polymer end groups at the termini of the copolymer. The quick kinetics of the reverse thermal gelation and the harmless character of the Michael-type addition between two sets of terminal groups, acrylates on one set and thiols on the other, allows irreversibly cross-linked hydrogels to be obtained in a rapid and biocompatible fashion, even when gelation was conducted in direct contact with cells. This allows in principle for an application of the tandem process in cell encapsulation. In the present work, we have optimized the macromolecular architecture and functionality of the precursors for allowing the use of the tandem process in encapsulation devices designed for calcium alginate. The mechanical, diffusional and biocompatibility properties of these materials were characterized; the comparison of mass transport properties of the tandem gels with those of calcium alginate suggests a similar or even better immunoisolation effect. PMID- 15109836 TI - Biological responses to cationically charged phosphorylcholine-based materials in vitro. AB - Phosphorylcholine (PC)-based polymers have been used in a variety of medical device applications to improve biocompatibility. The use of PC-based materials for biomaterials is associated with low protein adsorption, reduced complement activation, low inflammatory response and cell adhesion. For some medical device applications however, materials that support cell adhesion are also beneficial, allowing host interaction and encouraging full incorporation within the body. As previous studies have suggested that cell adhesion to materials is enhanced by the addition of charge, PC-based polymers have therefore been modified to incorporate various concentrations of cationic charge. In this study, the affect of cationic charge on a range of biological responses was investigated. In vitro assays have been used to assess the adsorption of protein onto the materials surface, the adhesion of mouse fibroblasts and rabbit corneal epithelial cells and the adhesion of human mononuclear cells and granulocytes. The results corroborate previous work showing that PC without charge significantly reduces protein adsorption, cell adhesion and inflammatory cell activation. The addition of cationic charge to PC polymers however, resulted in an increase in all of the above responses. This increase did not however, increase linearly with cationic monomer concentration. The differences in cell adhesion are discussed in terms of differences in protein adsorption, cytotoxicity and/or stability of the different cationic polymer coatings. PMID- 15109837 TI - Endothelialization of a non-woven silk fibroin net for use in tissue engineering: growth and gene regulation of human endothelial cells. AB - We have previously shown that a biomaterial consisting of a non-woven fibroin net produced from silk (Bombyx mori) cocoons is an excellent scaffolding material for a wide variety of human cells of different tissue types. Endothelialization must take place for a biomaterial to be successful after implantation. Therefore, primary human endothelial cells and the human endothelial cell lines, HPMEC ST1.6R and ISO-HAS-1, were examined for adherence and growth patterns on the fibroin nets by confocal laser scanning microscopy after vital staining of the cells and by electron microscopy. Endothelial cells adhered and spread along individual fibers of the nets and did not fill the gaps between individual fibers. Higher attachment and growth coverage was obtained if nets were first coated with gelatin, fibronectin or collagen type I. Proinflammatory markers of endothelial cells on the fibers exhibited a non-activated state and LPS stimulated cells exhibited activation of these markers. Furthermore, a typical PECAM-1 localization at cell-cell contacts was observed. Scanning electron microscopic examination of fibroin nets after removal of cells did not demonstrate any changes to the fibroin structure. HUVEC and HDMEC on fibroin nets embedded in collagen type I gels formed microvessel-like structures. Thus, silk fibroin nets are a highly endothelial cell-compatible scaffolding material that support the growth, normal and inducible cell functions and angiogenesis potential of human endothelial cells in vitro similar to that observed in vivo. PMID- 15109838 TI - Characterization and evaluation of chitosan matrix for in vitro growth of MCF-7 breast cancer cell lines. AB - Biodegradable polymer scaffolds were prepared from chitosan with varying degree of deacetylation for in vitro culture of human breast cancer MCF-7 cell lines. These polymers were characterized in terms of functional groups by FTIR and swelling properties. Polymers having high degree of deacetylation showed better swelling properties irrespective of the molecular weight. These polymers were biocompatible and non-toxic towards human epithelial MCF-7 cell lines. Attachment kinetics of MCF-7 cell lines on to polymer scaffold was investigated and it was observed that polymer having high degree of deacetylation favored better cell attachment. In CPIII polymer scaffold having 80% degree of deacetylation, a maximum of 1 millions cells per mg pf polymer were adsorbed within 1h. It appears that high swelling and high degree of deacetylation of chitosan helped in better adsorption of cancer cell lines. The cellular morphology of the attached cells on chitosan matrix was similar to that observed with regular plastic culture with the difference that, cells grew as three-dimensional clumps on chitosan matrix. Polymer having high degree of deacetylation not only favored better adsorption but also showed improved cell growth kinetics. Maximum cell concentration of 6.5 x 10(5) cells/ml was achieved in 5 days culture on CPIII polymer scaffold. The glucose consumption and lactate production pattern of the MCF-7 cell lines on chitosan polymer matrix were similar to that observed on cell growth on tissue culture flask. These results indicate that chitosan scaffold having high degree of deacetylation can be used for three-dimensional growth of MCF-7 cancer cell lines. Such in vitro 3D culture of cancer cells can thus be used as a model for the cytotoxic evaluation of anticancer drugs. PMID- 15109839 TI - Biodegradability of poly (2-hydroxyethyl methacrylate) in the presence of the J774.2 macrophage cell line. AB - The degradation of cross-linked and linear poly(2-hydroxyethyl methacrylate) (pHEMA), was examined in vitro with J774.2 cells. pHEMA microbeads were prepared with both types of polymers. Only cells in contact with the microbeads increased their production of lysosomal enzymes (TRAcP and ANAE) and released large amounts of reactive oxygen species with both types of pHEMA microbeads. Electron microscopy showed that macrophages were able to erode the surface of linear pHEMA but unable to erode the surface of the cross-linked polymer. Cells appeared wrapped by the linear pHEMA surface, but those cultured on the cross-linked polymer were only laying at the surface. After cell culture, the surface roughness of pHEMA slices was observed by atomic force microscopy (AFM). There was a significant increase in roughness (R(a)) of the surface of linear pHEMA slices cultured with J774.2 cells whereas no difference in R(a) between the surface of cross-linked pHEMA slices could be measured. AFM image of the hydrated materials were done: the surface of linear pHEMA swelled considerably in saline whereas the hydrated cross-linked polymer did not differ from the air-dried appearance. In conclusion, linear pHEMA swells in biological fluids, activates macrophages in close contact with the polymer and can be progressively eroded. PMID- 15109840 TI - Interpretation of electrical polarization and depolarization mechanisms of bioactive glasses in relation to ionic migration. AB - Electrical polarization and depolarization processes of the 45S5 type bioactive glass (BG) was studied by a thermally stimulated depolarization current (TSDC) and an complex impedance measurements. The depolarization processes were found to consist of three processes, which were observed as three peaks, P1, P2 and P3, in the TSDC spectra. P1 and P2, observed at lower than the glass transition temperature, were attributed to the sodium ionic migration from the interstitial positions after polarization procedure coordinated among neighboring silicate and phosphate to stabler positions. This migration process is presumed to relate to a shift in the ionic conductivity elucidated by the ac method. P3 observed at higher temperature for the BG polarized at more than 500 degrees Celsius for long period was ascribed to the ionic migration activated by a local deformation. The depolarization of BG consisted of three processes that depended on the neighboring coordination of traveling sodium ions during polarization. PMID- 15109841 TI - Biphasic calcium phosphate nanocomposite porous scaffolds for load-bearing bone tissue engineering. AB - A novel biodegradable nanocomposite porous scaffold comprising a beta-tricalcium phosphate (beta-TCP) matrix and hydroxyl apatite (HA) nanofibers was developed and studied for load-bearing bone tissue engineering. HA nanofibers were prepared with a biomimetic precipitation method. The composite scaffolds were fabricated by a method combining the gel casting and polymer sponge techniques. The role of HA nanofibers in enhancing the mechanical properties of the scaffold was investigated. Compression tests were performed to measure the compressive strength, modulus and toughness of the porous scaffolds. The identification and morphology of HA nanofibers were determined by X-ray diffraction and transmission electron microscopy, respectively. Scanning electron microscopy was used to examine the morphology of porous scaffolds and fracture surfaces to reveal the dominant toughening mechanisms. The results showed that the mechanical property of the scaffold was significantly enhanced by the inclusion of HA nanofibers. The porous composite scaffold attained a compressive strength of 9.8 +/- 0.3 MPa, comparable to the high-end value (2-10 MPa) of cancellous bone. The toughness of the scaffold increased from 1.00+/-0.04 to 1.72+/-0.02 kN/m, as the concentration of HA nanofibers increased from 0 to 5 wt %. PMID- 15109842 TI - Fabrication and characterizations of a novel drug delivery device liposomes-in microsphere (LIM). AB - In the present work, we developed a novel drug delivery system, liposomes-in microsphere (LIM) of biodegradable polymers, which is conceived from a combination of the polymer- and the lipid-based delivery systems and can thus integrate the advantages and avoid the drawbacks of the two systems. Liposomes were encapsulated into microspheres of biodegradable polymers by the solvent extraction/evaporation process to form LIMs. The integrity of the liposomes was preserved by modifying the microencapsulation process and coating the liposomes with chitosan. We demonstrated by scanning electron microscopy, laser light scattering and fluorescence spectroscopy that the particle size and surface morphology of the polymeric microspheres did not change significantly with the liposomes encapsulated, the liposomes remained intact within the polymeric matrix of the microspheres, and the encapsulated liposomes could be released from the microspheres in a controlled manner at a nearly constant release rate after an initial off-release period. Decreasing the particle size of liposomes and increasing the pore size of the polymeric matrix shortened the initial off release period and increased the liposome release rate. In conclusion, a novel drug delivery system, liposomes-in-microsphere, was successfully developed and characterized. The liposome release kinetics could be controlled by the composition and fabrication parameters of the liposomes and polymeric microspheres. Such a novel controlled release system may have potential to be applied for drug delivery and gene therapy. PMID- 15109843 TI - The effect of simulated body fluid on the mechanical properties of multiblock poly(aliphatic/aromatic-ester) copolymers. AB - The effects of simulated body fluid (SBF) on the surface and mechanical properties of poly(aliphatic/aromatic-ester) (PED) copolymers were investigated. PED copolymers containing different hard (aromatic) and soft (aliphatic) segment weight ratios were exposed to SBF for 52 days, and afterwards their mechanical properties were evaluated. A quasi-static tensile test estimating relaxation and fatigue properties from the hysteresis method was performed. The surface properties were monitored with scanning electron microscopy and energy-dispersive X-ray spectrometry. PED copolymers showed various susceptibilities to apatite layer formation on the surface. This was ascribed to the hard/soft segment ratios of PED materials. Polymers containing higher amounts of amorphous soft segments showed a better deposition with the apatite layer, which provided improved mechanical properties, especially fatigue (improved dynamic creep resistance). PMID- 15109844 TI - Bone bonding to hydroxyapatite and titanium surfaces on femoral stems retrieved from human subjects at autopsy. AB - The success of clinical results obtained with many hydroxyapatite (HA)-coated prosthetic designs has deflected attention from the need to extend the life of the HA coating on the device. In the current study the percentages of HA and titanium surfaces to which bone was bonded, on HA-coated and non-coated titanium femoral stems retrieved from human subjects, were evaluated. Plasma-sprayed hydroxyapatite (PSHA)-coated devices demonstrated wide variability in the percentage of the PSHA coating remaining on the stems. The coating was missing from a substantial portion of a stem after only about 6 months of implantation. The percentage of revealed metal to which bone was bonded was significantly less than the percentage of the HA coating demonstrating such bonding. The revealed metal to which bone was bonded was comparable to the same value for a separate group of non-PSHA-coated titanium stems. If HA-coatings degrade over time precipitous decline in performance may occur even after several functional years. Many ultrastructural features of the bone bonded to the HA coatings on these implants from human subjects were comparable to those found on HA-coated devices implanted in a canine model. PMID- 15109845 TI - Transcatheter embolization using degradable crosslinked hydrogels. AB - Therapeutic embolization is the selective transcatheter blockage of blood vessels or diseased vascular structures. The majority of current embolization materials in clinical use are permanent. There are clinical situations however, in which temporary embolization is desired. Degradable hydroxyethyl acrylate (HEA) microspheres have been synthesized. Canine renal arteries and rabbit central auricular arteries were embolized with HEA microspheres, and compared with degradable human serum albumin (HSA) microspheres, and permanent microspheres. HSA and HEA microspheres both achieved temporary occlusions. HSA and HEA microspheres were recanalizated at 1 and 3 weeks, respectively, while arteries occluded with permanent microspheres did not recanalize. All embolic microspheres led to tissue infarction, with the short-term HSA microspheres providing the least damage, and the permanent microspheres leading to extensive damage. Advantages of temporary embolization were not convincingly demonstrated since temporary occlusions still led to tissue infarction. PMID- 15109847 TI - Surface-MALDI mass spectrometry in biomaterials research. AB - Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) has been used for over a decade for the determination of purity and accurate molecular masses of macromolecular analytes, such as proteins, in solution. In the last few years the technique has been adapted to become a new surface analysis method with unique capabilities that complement established biomaterial surface analysis methods such as XPS and ToF-SSIMS. These new MALDI variant methods, which we shall collectively summarize as Surface-MALDI-MS, are capable of desorbing adsorbed macromolecules from biomaterial surfaces and detecting their molecular ions with high mass resolution and at levels much below monolayer coverage. Thus, Surface-MALDI-MS offers unique means of addressing biomaterial surface analysis needs, such as identification of the proteins and lipids that adsorb from multicomponent biological solutions in vitro and in vivo, the study of interactions between biomaterial surfaces and biomolecules, and identification of surface-enriched additives and contaminants. Surface-MALDI-MS is rapid, experimentally convenient, overcomes limitations in mass resolution and sensitivity of established biochemical techniques such as SDS-PAGE, and can in some circumstances be used for the quantitative analysis of adsorbed protein amounts. At this early stage of development, however, limitations exist: in some cases proteins are not detectable, which appears to be related to tight surface binding. This review summarizes ways in which Surface-MALDI-MS methods have been applied to the study of a range of issues in biomaterials surfaces research. PMID- 15109848 TI - Regulation of osteoclast activity in peri-implant tissues. AB - Implants, particularly joint replacement prostheses, are one of the great success stories of modern medicine. However, too many implants fail prematurely, mainly due to aseptic bone loss around the implant. This paper reviews our current understanding of the role of osteoclasts in this peri-implant bone lysis. Prosthetic particles, often produced by articulating prostheses, are one of the major causes of elevated osteoclast lysis of peri-implant bone. Over the past decade there have been major advances in our understanding of the factors that regulate osteoclast activity, many of which were found to be important in osteoclast formation and activity in the peri-implant tissues. These factors are targets of a number of recently developed drugs that have been used successfully to prevent and treat peri-implant bone lysis in experimental models. Treatments such as these are being used in a number of bone loss pathologies in humans and have the potential for successful treatment of peri-implant osteolysis. In addition, understanding how different biomaterials influence the expression of key osteoclastogenic factors may allow us to select biomaterials for implantation that will last the lifetime of the recipient. PMID- 15109849 TI - Long-term in vivo biostability of poly(dimethylsiloxane)/poly(hexamethylene oxide) mixed macrodiol-based polyurethane elastomers. AB - The long-term biostability of a novel thermoplastic polyurethane elastomer (Elast Eon 2 80A) synthesized using poly(hexamethylene oxide) (PHMO) and poly(dimethylsiloxane) (PDMS) macrodiols has been studied using an in vivo ovine model. The material's biostability was compared with that of three commercially available control materials, Pellethane 2363-80A, Pellethane 2363-55D and Bionate 55D, after subcutaneous implantation of strained compression moulded flat sheet dumbbells in sheep for periods ranging from 3 to 24 months. Scanning electron microscopy, attenuated total reflectance-Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy were used to assess changes in the surface chemical structure and morphology of the materials. Gel permeation chromatography, differential scanning calorimetry and tensile testing were used to examine changes in bulk characteristics of the materials. The results showed that the biostability of the soft flexible PDMS-based test polyurethane was significantly better than the control material of similar softness, Pellethane 80A, and as good as or better than both of the harder commercially available negative control polyurethanes, Pellethane 55D and Bionate 55D. Changes observed in the surface of the Pellethane materials were consistent with oxidation of the aliphatic polyether soft segment and hydrolysis of the urethane bonds joining hard to soft segment with degradation in Pellethane 80A significantly more severe than that observed in Pellethane 55D. Very minor changes were seen on the surfaces of the Elast-Eon 2 80A and Bionate 55D materials. There was a general trend of molecular weight decreasing with time across all polymers and the molecular weights of all materials decreased at a similar relative rate. The polydispersity ratio, Mw/Mn, increased with time for all materials. Tensile tests indicated that UTS increased in Elast-Eon 2 80A and Bionate 55D following implantation under strained conditions. However, ultimate strain decreased and elastic modulus increased in the explanted specimens of all three materials when compared with their unimplanted unstrained counterparts. The results indicate that a soft, flexible PDMS-based polyurethane synthesized using 20% PHMO and 80% PDMS macrodiols has excellent long-term biostability compared with commercially available polyurethanes. PMID- 15109850 TI - Bone formation within alumina tubes: effect of calcium, manganese, and chromium dopants. AB - Alumina tubes (1.3mm outer diameter, 0.6mm inner diameter, 15 mm length) doped with Ca, Mn, or Cr at nominal concentrations of 0.5 and 5.0 mol% were implanted into femoral medullary canals of female rats for 16 weeks. Tissue formation within tubes was determined by histology and histomorphometry. Addition of Ca to alumina promoted hypertrophic bone formation at the advancing tissue fronts and tube entrances, and appeared to retard angiogenesis by limiting ongoing cellular migration into the tube. It is speculated that the presence of a secondary phase of calcium hexaluminate, probably having a solubility greater than that of alumina, possibly increased the level of extracellular Ca and, consequently, stimulated osteoclastic activity at the bone-ceramic interface. Addition of Mn significantly enhanced osteogenesis within the tubes. However, it is not possible to determine whether phase composition or microstructure of the ceramic was responsible for this because both were significantly altered by Mn addition. Addition of Cr to the alumina apparently stimulated bone remodelling as indicated by increased cellular activity and bone resorption at the tissue-implant interface. Cr was incorporated into the alumina as a solid solution and the tissue response was speculated to be an effect of surface chemistry rather than microstructure. The work demonstrates that doping a bioinert ceramic with small amounts of specific elements can significantly alter tissue ingrowth, differentiation, and osteogenesis within a porous implant. PMID- 15109851 TI - The modulation of osteogenesis in vitro by calcium titanium phosphate coatings. AB - Calcium phosphate coated titanium and titanium alloy are widely used as dental and orthopaedic implants. This study examines the effect of novel calcium titanium and calcium titanium zirconium phosphates suitable for plasma-spraying onto titanium substrata on the expression of bone-related genes and proteins by human bone-derived cells (HBDC) and compares this behavior to that on native titanium and hydroxyapatite-coated titanium. Test materials were an acid etched and sand-blasted titanium surface (Ti-DPS), a plasma-sprayed hydroxyapatite coating (HA), and five materials which were created from CaTi(4)(PO(4))(6) (CTP) and CaZr(4)(PO(4))(6) (CZP): sintered CaTi(4)(PO(4))(6) (CTP-S1), sintered 46CaO.23TiO(2).31P(2)O(5) (CTP-S2), sintered CaTiZr(3)(PO(4))(6), (CTZP-S1), sintered 46CaO.23ZrO(2).31P(2)O(5) (CTZP-S2) and sintered 55CaO.20TiO(2).31P(2)O(5) (CTP-S3). HBDC were grown on the substrata for 3, 7, 14 and 21 d, counted and probed for various mRNAs and proteins (type I collagen, osteocalcin, osteopontin, osteonectin, alkaline phosphatase and bone sialoprotein). All substrates significantly affected cellular growth and the temporal expression of an array of bone-related genes and proteins. At 14 and 21 d, cells on CTP-S3 displayed significantly enhanced expression of all osteogenic mRNAs. Surfaces of CTP-S1 and CTP-S3 had the most effect on osteoblastic differentiation inducing a greater expression of an array of osteogenic markers than recorded for cells grown on Ti-DPS and HA, suggesting that these novel materials may possess a higher potency to enhance osteogenesis. PMID- 15109852 TI - Synthetic elastin hydrogels derived from massive elastic assemblies of self organized human protein monomers. AB - A key objective of bioengineering is the development of new scaffolding biomaterials with appropriate mechanical and biological properties such as strength, elasticity and biocompatibility that mimic the native host connective tissue. Here we describe the production and properties of massive synthetic elastin assemblies formed by chemically cross-linking recombinant human tropoelastin with bis(sulfosuccinimidyl) suberate, permitting the construction of elastic sponges, sheets and tubes. The innate characteristics of synthetic elastin constructs are common with those of native elastin. The Young's Modulus ranged from 220 to 280 kPa with linearity of extension to at least 150%. Synthetic elastin was extensible by 200-370%. The constructs behaved as hydrogels and displayed stimuli-responsive characteristics towards temperature and salt concentrations. Intrinsic fluorescence spectroscopy demonstrated that the elastin fluorophore is a feature of the polypeptide. Scanning electron microscopy allowed us to construct a model of elastin assembly that was driven by the lateral association of small twisted rope-like fibrils. FT-Raman spectra at 100% strain gave amide I and III peaks that correlated with a stretch-dependent increase in alpha-helical content. Growth and proliferation of cells were supported in vitro while in vivo implants were well tolerated. We conclude that synthetic elastin has potential as a novel biomaterial that can be easily molded into a variety of shaped tissue substrates and has a range of properties that are required for elastic, cell-interacting and compliant applications. Furthermore, its in vitro construction provides a powerful tool to probe the early stages of elastin assembly and the molecular basis for its elasticity. PMID- 15109853 TI - Cemented fixation with PMMA or Bis-GMA resin hydroxyapatite cement: effect of implant surface roughness. AB - Implant surface roughness is an important parameter governing the overall mechanical properties at the implant-cement interface. This study investigated the influence of surface roughness using polymethylmethcrylate (PMMA) and a Bisphenol-a-glycidylmethacyrlate resin-hydroxyapatite cement (CAP). Mechanical fixation at the implant-cement interface was evaluated in vitro using static shear and fatigue loading with cobalt chrome alloy (CoCr) dowels with different surface roughness preparations. Increasing surface roughness improved the mechanical properties at the implant-cement interface for both types of cement. CAP cement fixation was superior to PMMA under static and dynamic loading. PMID- 15109854 TI - Influence of fluorapatite on the properties of thermally sprayed hydroxyapatite coatings. AB - Thermally sprayed hydroxyapatite has been the widely used on orthopaedic prosthesis to induce bone growth and facilitate bone attachment. However, hydroxyapatite has a greater affinity for the formation of an amorphous phase in the thermally sprayed coating that results in the release of excessive amount of mineral ions from the implant coating leading to a saturated environment in the immediate vicinity of the bone cells. Fluorapatite however is highly crystalline and offers the potential for lower mineral ion release by dissolution. Thus study investigates the influence of fluorapatite in a thermally sprayed hydroxyapatite coating. Mechanical blends of fluorapatite with hydroxyapatite were thermally sprayed, characterized with X-ray diffraction, SEM, FTIR, optical microscopy for microstructure, roughness and tested for solubility. Cathodoluminescence microscopy was used to examine the resorbed coating surface. Fluorapatite coatings crystallized more readily and produce a greater coating roughness. The roughness in fluorapatite coatings arises from less flattened droplets that show a tendency for finger formation. Addition of fluorapatite increases coating crystallinity. The use of slower resorbing fluorapatite produces less particle release which favors improved osseointegration. Less change in the surface topography during resorption can be used to an advantage to control the coating surface presented to cells and extra cellular matrix proteins. PMID- 15109855 TI - Analysis of 3D bone ingrowth into polymer scaffolds via micro-computed tomography imaging. AB - This paper illustrates the utility of micro-computed tomography (micro-CT) to study the process of tissue engineered bone growth. A micro-CT facility for imaging and visualising biomaterials in three dimensions (3D) is described. The facility is capable of acquiring 3D images made up of 2000(3) voxels on specimens up to 60mm in extent with resolutions down to 2 microm. This allows the 3D structure of tissue engineered materials to be imaged across three orders of magnitude of detail. The capabilities of micro-CT are demonstrated by imaging the Haversian network within human femoral cortical bone (distal diaphysis) and bone ingrowth into a porous scaffold at varying resolutions. Phase identification combined with 3D visualisation enables one to observe the complex topology of the canalicular system of the cortical bone. Imaging of the tissue engineered bone at a scale of 1cm and resolutions of 10 microm allows visualisation of the complex ingrowth of bone into the polymer scaffold. Further imaging at 2 microm resolution allows observation of bone ultra-structure. These observations illustrate the benefits of tomography over traditional techniques for the characterisation of bone morphology and interconnectivity and performs a complimentary role to current histomorphometric techniques. PMID- 15109856 TI - Biodegradable composite scaffolds with an interconnected spherical network for bone tissue engineering. AB - Tissue engineering scaffolds are highly engineered structures that accommodate cells, facilitate their expression, and resorb to facilitate regeneration of tissue. A new technique for producing controlled pore shape and pore size interconnectivity offers promise for application as a tissue engineering scaffold. Salt particles were spheroidized in a flame and sintered to provide an interconnecting salt template. The salt template was filled with a carbonated fluorapatite powder and a polylactic polymer to produce a composite scaffold. It was found that a higher pore space is possible with the use of spherical and larger salt particle sizes. This technique can produce scaffolds with good interconnectivity and be suitable for producing pore size graded bodies. PMID- 15109857 TI - Colonization and maintenance of murine embryonic stem cells on poly(alpha-hydroxy esters). AB - The aim of this study was to determine the ability of various poly(alpha-hydroxy esters) to support the in vitro propagation of murine embryonic stem (ES) cells in an undifferentiated state. To this end, ES cell colonization, growth and Oct-4 immunoreactivity following a 48 h culture period upon poly((D,L)-lactide), poly((L)-lactide), poly(glycolide) and poly((D,L)-lactide-co-glycolide) (PLGA) were assessed. By the analysis of live and dead cell number indices and Oct-4 immunoreactivity, ES cell colonization rate during a 48 h culture period was found to be significantly greater on PLGA compared to all the other unmodified poly(alpha-hydroxy esters) tested. Surface treatment of all polymers with 0.1m potassium hydroxide revealed a significant increase in ES cell live numbers when compared to all unmodified polymers, thus revealing a correlation between polymer content, hydrophilicity and colonization rate. These data suggest that surface treated poly(alpha-hydroxy esters) may be employed for ES cell scale up procedures and in tissue engineering applications requiring the colonization of scaffolds by ES cells in an undifferentiated state. According to such applications, once the designated scaffold has been colonized, ES cell directed differentiation into the desired and fully differentiated, functional adult tissue may then be effected. PMID- 15109858 TI - Morphology of sol-gel derived nano-coated coralline hydroxyapatite. AB - Current bone graft materials are mainly produced from coralline hydroxyapatite (HAp). Due to the nature of the conversion process, commercial coralline HAp has retained coral or CaCO(3,) and the structure possesses nanopores within the inter pore trabeculae, resulting in high dissolution rates. Under certain conditions these features reduce durability and strength and are not utilised where high structural strength is required. To overcome these limitations, a new coral double-conversion technique has been developed. The technique involves a two stage application route where, in the first stage, complete conversion of coral to pure HAp is achieved. In the second, a new sol-gel-derived HAp nano-coating is directly applied to cover the micro- and nano-pores within the intra-pore material, whilst maintaining the large pores. Biaxial strength was improved two fold due to this unique double treatment. This application is expected to result in enhanced durability and longevity due to the monophasic hydroxyapatite structure and strength in the physiological environment. It is anticipated that this new material can be applied to load-bearing bone graft applications where high strength requirements are pertinent. PMID- 15109859 TI - Sintered hydroxyfluorapatites--IV: The effect of fluoride substitutions upon colonisation of hydroxyapatites by mouse embryonic stem cells. AB - Biodegradable scaffolds serve a central role for tissue engineering scaffolds and guiding tissue regeneration. Some of these scaffolds, including apatites, display a significant effect upon cell adhesion and cell proliferation. The incorporation of scaffold technology with the developing embryonic stem (ES) cell field and the capacity of ES cells for self-renewal and differentiation are believed to hold enormous potential for applications in biomedical research and regenerative medicine. The purpose of this work was to determine the effect of hydroxyapatite (HAP) and fluoride substitutions of HAP upon ES cell growth and colonisation. Sintered hydroxyfluorapatite discs were found to support cellular proliferation and colonisation, and the ES cells displayed a tendency for differentiation on the apatite surface as determined by reductions in colony Oct4 immunoreactivity. Fluoride-containing HAPs were found to provide equivalent support to gelatin in terms of cell numbers, yet superior support for cellular colonisation when compared to HAP. This study indicates that fluoride substitutions of HAP may represent a viable strategy for the development of certain engineered tissue replacements and tissue regeneration systems using ES cells. PMID- 15109860 TI - Fibre reinforced composite dental bridge. Part I: Experimental investigation. AB - This experimental investigation aims at revealing the mechanical behaviour and failure pattern of direct fibre-reinforced resin-bonded dental bridge with various designs. To evaluate the overall effects of some newly developed dental materials, in the experiment, genuine composite dental bridge specimens are prepared and tested. The ultimate load, stiffness and mode at the failure of the bridges are measured and compared with the design variations. A good agreement between test and some clinical observations is demonstrated. It is verified that the weakest region appears across the pontic-abutment interface in the composite bridges. This study suggests that the composite bridges reinforced by fibres and supported by adjacent teeth could be of a higher structural strength and stiffness; therefore would provide better clinical performances. PMID- 15109861 TI - Fibre reinforced composite dental bridge. Part II: Numerical investigation. AB - Motivated by the clinical success and limitations on experimental investigation of the fibre-reinforced composite dental bridge, this paper aims at providing a numerical investigation into the bridge structure. The finite element (FE) model adopted here is constructed from computer tomography images of a physical bridge specimen. The stress and strain distributions in the bridge structure especially in the bonding interfaces are analyzed in detail. The peak stresses and their variations with the different bridge designs are evaluated. Due to the lower bond strengths of adhesives and the high stress concentration in the pontic-abutment interface, the likelihood of failure in the interface is predicted by finite element analysis. The validity of the numerical results is established by a good agreement between the FE prediction and the tests in the load-deflection responses, the structural stiffness as well as the failure location of the composite dental bridge. PMID- 15109862 TI - Furanones as potential anti-bacterial coatings on biomaterials. AB - A major barrier to the long-term use of medical devices is development of infection. Staphylococcus epidermidis is one of the most common bacterial isolates from these infections with biofilm formation being their main virulence factor. Currently, antibiotics are used as the main form of therapy. However with the emergence of staphylococcal resistance, this form of therapy is fast becoming ineffective. In this study, the ability of a novel furanone antimicrobial compound to inhibit S. epidermidis adhesion and slime production on biomaterials was assessed. Furanones were physically adsorbed to various biomaterials and bacterial load determined using radioactivity. Slime production was assessed using a colorimetric method. Additionally, the effect of the furanone coating on material surface characteristics such as hydrophobicity and surface roughness was also investigated. The results of this study indicated that there was no significant change in the material characteristics after furanone coating. Bacterial load on all furanone-coated materials was significantly reduced (p<0.001) as was slime production (p<0.001). There is a potential for furanone coated biomaterials to be used to reduce medical device-associated infections. PMID- 15109863 TI - Biological performance of a novel synthetic furanone-based antimicrobial. AB - Infection of medical devices causes significant morbidity and mortality and considerable research effort has been directed at solving this problem. The aim of this study was to assess the biological performance of a novel furanone compound that has potential as an anti-infective coating for medical devices. This study examined in vitro leukocyte response following exposure to the antibacterial 3-(1'-bromohexyl)-5-dibromomethylene-2(5H)-furanone and assessed the tissue response following subcutaneous implantation of the furanone compound covalently bound to polystyrene (PS). Peripheral human blood was exposed to furanones in solution for 1h and flow cytometry used to analyse viability and changes in expression of surface receptors CD11b/CD18 and CD44. Flow cytometry results from propidium iodide stained cell suspensions suggested that the leukocytes were viable after exposure to furanones in whole blood. No significant difference was found in the expression of CD11b/CD18 and CD44 between the furanone exposed samples and the negative control for neutrophils suggesting that the furanones themselves do not activate these leukocytes. The positive control lipopolysaccharide significantly up-regulated CD11b/CD18 and slightly down regulated CD44 on both PMNs and monocytes. In vivo studies of the tissue response to furanone covalently bound to PS showed that there was no significant difference in cellularity of capsules surrounding the disk and no significant increase in myeloperoxidase expression. These results demonstrate negligible acute inflammatory response to synthetic brominated antibacterial furanones. Future studies will focus on chronic responses and examination of in vivo efficacy. PMID- 15109864 TI - The control of Staphylococcus epidermidis biofilm formation and in vivo infection rates by covalently bound furanones. AB - In order to overcome the continuing infection rate associated with biomaterials, the use of covalently bound furanones as an antibiofilm coating for biomaterials has been investigated. Furanones have previously been shown to inhibit growth of Gram-positive and Gram-negative bacteria. The aim of these studies were to covalently bind furanones to polymers and to test their efficacy for inhibiting biofilm formation of Staphylococcus epidermidis and in vivo infection rate. Two methods of covalent attachment of furanones were used. The first, a co polymerisation with a styrene polymer, and second, a plasma-1-ethyl-3 (dimethylaminopropyl) carbodiimide (EDC) reaction to produce furanone-coated catheters. Biofilm formation by S. epidermidis in vitro was inhibited by 89% for polystryene-furanone disks and by 78% by furanone-coated catheters (p<0.01). In an in vivo sheep model we found furanones were effective at controlling infection for up to 65 days. Furanones have potential to be used as a coating for biomaterials to control infection caused by S. epidermidis. PMID- 15109865 TI - Analysis of tensile bond strengths using Weibull statistics. AB - Tensile strength tests of restorative resins bonded to dentin, and the resultant strengths of interfaces between the two, exhibit wide variability. Many variables can affect test results, including specimen preparation and storage, test rig design and experimental technique. However, the more fundamental source of variability, that associated with the brittle nature of the materials, has received little attention. This paper analyzes results from micro-tensile tests on unfilled resins and adhesive bonds between restorative resin composite and dentin in terms of reliability using the Weibull probability of failure method. Results for the tensile strengths of Scotchbond Multipurpose Adhesive (3M) and Clearfil LB Bond (Kuraray) bonding resins showed Weibull moduli (m) of 6.17 (95% confidence interval, 5.25-7.19) and 5.01 (95% confidence interval, 4.23-5.8). Analysis of results for micro-tensile tests on bond strengths to dentin gave moduli between 1.81 (Clearfil Liner Bond 2V) and 4.99 (Gluma One Bond, Kulzer). Material systems with m in this range do not have a well-defined strength. The Weibull approach also enables the size dependence of the strength to be estimated. An example where the bonding area was changed from 3.1 to 1.1 mm diameter is shown. Weibull analysis provides a method for determining the reliability of strength measurements in the analysis of data from bond strength and tensile tests on dental restorative materials. PMID- 15109866 TI - In vivo evaluation of resorbable bone graft substitutes in a rabbit tibial defect model. AB - Calcium sulfate as a bone graft substitute is rapidly resorbed in vivo releasing calcium ions but fails to provide long-term three-dimensional framework to support osteoconduction. The setting properties of calcium sulfate however allow it to be applied in a slurry form making it easier to handle and apply in different situations. This study examines the in vivo response of calcium sulfate alone and as a carrier for a coralline hydroxyapatite in an established bilateral corticocancellous defect model in rabbits. Defects were filled flush to the anterior cortex with a resorbable porous ceramic alone and in combination with calcium sulfate slurry, calcium sulfate slurry alone or calcium sulfate pellets and examined at time points up to 52 weeks. Specimens where assessed using Faxitron X-ray, light and electron microscopy. Calcium sulfate in either slurry or pellet form does indeed support new bone formation alone however, complete filling of the bone defect is not observed. Calcium sulfate in slurry form does however improve the surgical handling of particulate bone graft substitutes such as Pro Osteon 200 R, which remained as an osteoconductive scaffold for up to 52 weeks and may have played an important role in the ultimate closure of the cortical windows. PMID- 15109867 TI - Strength, reliability and mode of fracture of bilayered porcelain/zirconia (Y TZP) dental ceramics. AB - The aim of this study was to investigate the biaxial flexural strength, reliability and the mode of fracture of bilayered porcelain/zirconia (Y-TZP) disks. For this purpose, 80 specimens were made from conventional dental porcelain and Y-TZP core ceramic, and equally divided into four groups as follows: monolithic specimens of porcelain; monolithic specimens of core material; bilayered specimens with the porcelain on top (facing the loading piston during testing); bilayered specimens with core material on top. The maximum load at fracture was calculated with a biaxial flexural test and finite element analysis was used to estimate the maximum tensile stress at fracture. Results were analyzed with one-way ANOVA, Tukey HSD. The reliability of strength was analyzed with the Weibull distribution. SEM was used to identify the initial crack and characterize the fracture mode. Monolithic core specimens and bilayered sample with the core material on the bottom were statistically significantly stronger than monolithic porcelain disks and bilayered samples with the porcelain on the bottom. The study, which was conducted with sample configurations that reproduce the clinical situation of crowns and fixed partial dentures, indicates that the material which lies on the bottom surface dictates the strength, reliability and fracture mode of the specimens. The contribution of strong and tough core materials to the performance of all-ceramics restorations may be offset by the weaker veneering porcelain if the actual distribution of the tensile stresses within the restoration is not taken into consideration. PMID- 15109868 TI - Gravity spun polycaprolactone fibres: controlling release of a hydrophilic macromolecule (ovalbumin) and a lipophilic drug (progesterone). AB - A hydrophilic macromolecule (ovalbumin (OVA)) and a lipophilic drug (progesterone) were incorporated in polycaprolactone (PCL) fibres by gravity spinning using particulate dispersions and co-solutions of PCL and steroid, respectively. PCL fibres loaded with 1% (w/w) OVA powder displayed a pronounced burst release phase (60% of the protein load) over 2 days in PBS at 37 degrees C. The release profile then tended to plateau. In contrast, OVA nanoparticle-loaded fibres exhibited delayed protein release initially and then a major increase at day 14. This behaviour may be useful for sequential release of polypeptide growth factors which are influential at specific time points in the wound healing process. SDS-PAGE analysis revealed that the protein molecular weight was conserved during fibre spinning. The amount of progesterone release from PCL fibres in PBS increased with drug loading but the cumulative release profiles (% w/w) were little affected by the initial drug loading of the fibres (1.5 and 3.5% w/w) or the concentration of the PCL spinning solution (12.5 and 20% w/v). Steroid delivery was rapid due to the high fibre surface area and high permeability of PCL resulting in complete drug loss over 24h. Released progesterone inhibited the growth of MCF-7 breast epithelial cells in culture, demonstrating retention of bioactivity. Gravity spinning shows potential for producing PCL fibre-based platforms for programmed delivery of bioactive molecules of utility for tissue engineering and drug delivery. PMID- 15109869 TI - NMR studies of a novel calcium, phosphate and fluoride delivery vehicle-alpha(S1) casein(59-79) by stabilized amorphous calcium fluoride phosphate nanocomplexes. AB - The repair of early tooth enamel lesions has been recently demonstrated by tryptic phosphopeptides derived from milk caseins that associate with amorphous calcium phosphate (ACP) forming stable complexes. These casein phosphopeptides (CPP), containing the cluster sequence-Ser(P)-Ser(P)-Ser(P)-Glu-Glu-, form calcium phosphate delivery vehicles that retard enamel demineralization and promote remineralization. Recently, we have shown that these peptides also stabilize calcium fluoride phosphate as soluble complexes. These complexes designated CPP-ACFP, have the potential to provide superior clinical efficacy in preventing dental caries and treating and repairing early stages of disease. In an approach to determine the ultrastructure of the casein phosphopeptide amorphous calcium fluoride phosphate complexes, we have studied the structure of the predominant peptide alpha(S1)-CN(59-79) bound to ACFP using nuclear magnetic resonance (NMR) spectroscopy and X-ray diffraction. The alpha(S1)-CN(59-79) peptide stabilized calcium fluoride phosphate as amorphous nanocomplexes with a hydrodynamic radius of 2.12+/-0.26 nm. The nanocomplexes exhibited stoichiometry of one peptide to 15 calcium, nine phosphate and three fluoride ions. Sequence specific resonance assignments were determined for the peptide alpha(S1)-CN(59 79) complexed to the ACFP. The secondary structure of the peptide alpha(S1)-CN(59 79) was characterized by sequential (i, i+1), medium-range (i, i+2) nOes and H alpha chemical shifts. The spectral data were compared with that of the peptide alpha(S1)-CN(59-79) bound to calcium ions, revealing that the structurally significant secondary NH and alpha-chemical shifts were similar. PMID- 15109870 TI - Drug release characteristics of phase separation pHEMA sponge materials. AB - A number of phase separation pHEMA sponge hydrogels have been prepared based on variations in monomer contents, concentration of cross-linking agent, solvent mixture and temperature of polymerization. The loading levels and release profiles of the anti-inflammatory drug prednisolone were examined for each of the pHEMA sponge materials. An effective diffusion coefficient determined by an optimization approach based on the experimental data was used to measure their release characteristics. The effect of morphological variations, revealed by the environmental scanning electron microscopy, and polymer/solvent volume fractions on these properties were discussed. PMID- 15109871 TI - A micro-mechanics model of dentin mechanical properties. AB - Application of a micro-mechanics cell model to dentin composites for determination of their effective mechanical properties is discussed in this paper. The dilute micro-mechanics model for fibre-reinforced composites is utilized and the corresponding cell model is chosen to consist of a circular hollow cylinder filled with liquid or gas phase, which is surrounded by two circular cylindrical shells, a thin shell and a matrix phase. Each layer of cylindrical shell is here considered as a composite consisting of collagen fibrils, with mineralized hydroxyapatite, loosely connected to their neighbours, and water (or gas in the case of dry dentin composite). Determination of the effective material properties of such a three phase composite is discussed. Using the cell model the effect of porosity, thickness of each cylindrical shell, and mineral content on material properties is analysed. Results obtained from nano indentation observations are compared with numerical predictions of the analytical model. PMID- 15109872 TI - Mechanical properties and microstructure of hypomineralised enamel of permanent teeth. AB - Isolated enamel defects are commonly seen in first permanent molar teeth but there has been little work on the physical and morphological composition of affected molars. The aim of this study was to determine the mechanical and morphological properties of hypomineralised first permanent molar teeth, utilising the Ultra-Micro-Indentation System (UMIS) and scanning electron microscope, respectively. Further investigations using Energy Dispersive X-ray Spectrometery (EDS), Back Scatter Electron (BSE) Imaging, and X-ray diffraction were employed to attempt to determine the chemical composition, mineral content and crystalline structure of the hypomineralised tissue, respectively, of eight first permanent molars with severe enamel hypomineralisation. The hardness and modulus of elasticity were found to be statistically significantly lower (0.53+/ 0.31 and 14.49+/-7.56 GPa, respectively) than normal enamel (3.66+/-0.75 and 75.57+/-9.98 GPa, respectively). Although the fractured surface of the hypomineralised enamel was significantly more disorganised and the relative mineral content was reduced by approximately 5% in comparison to sound enamel, the mineral phase and Ca/P ratio was similar in hypomineralised and sound enamel. The dramatic reduction in the mechanical properties of first permanent molar teeth has ramifications when clinicians are choosing restorative materials to restore the defects. The reason for the dramatic reduction in mechanical properties of hypomineralised first permanent molar teeth is at present unknown. PMID- 15109873 TI - Seasonal flux of nonylphenol in Han River, Korea. AB - In order to understand the behavior of nonylphenol (NP) in Han River, water, suspended particle and sediment samples were analyzed during summer, autumn and winter. Concentrations of nonylphenol in water ranged from 23.2 to 187.6 ng/l, in suspended particle from 6.8 to 190.8 ng/l and in sediment from 25.4 to 932.0 ng/gdrywt. An increasing trend in the concentration is noticed in all matrices along down the river. In case of water and suspended particle, concentrations were higher in warmer season than in colder season. Percentage of nonylphenol in the suspended particle phase decreased from 67% to 28% with decreasing temperature in water. A reasonable correlation (R2 = 0.63) was obtained for water and suspended particle. The partition coefficient Log Kp is 4.8. No seasonal variation of the concentration in sediment is noticed in this study. PMID- 15109874 TI - Fugacity ratio estimations for high-melting rigid aromatic compounds. AB - Prediction of the environmental fate of organic compounds requires knowledge of their tendency to stay in the gas and water phase. Vapor pressure and aqueous solubility are commonly used descriptors for these processes. Depending on the type of distribution process, values for either the pure solid state or the (subcooled) liquid state have to be used. Values for the (subcooled) liquid state can be calculated from those for the solid state, and vice versa, using the fugacity ratio. Fugacity ratios are usually calculated from the entropy of fusion and the melting point. For polycyclic aromatic hydrocarbons, chlorobenzenes, chlorodibenzofuranes, and chlorodibenzo(p)dioxins, fugacity ratios calculated using experimental entropies of fusion were systematically less than those obtained from a thermodynamically more rigorous approach using heat capacity data. The deviation was more than 1 order of magnitude at the highest melting point. The use of a universal value for the entropy of fusion of 56 J/molK resulted in either over or underestimation by up to more than 1 order of magnitude. A simple correction factor, based on the melting point only, was derived. This correction factor allowed the fugacity ratios to be estimated from experimental entropies of fusion and melting point with an accuracy better than 0.1-0.2 log units. PMID- 15109875 TI - The contribution of alkali soluble (humic acid-like) and unhydrolyzed-alkali soluble (core-humic acid-like) fractions extracted from maize plant to the formation of soil humic acid. AB - Alkali soluble (humic acid-like material) (HA-like) (yield of 132 gkgdm(-1)) and the unhydrolized-alkali soluble (core-humic acid-like material) (core-HA-like) (yield of 33.4 gkgdm(-1)) fractions were extracted from maize plants and characterized by C and N determinations, DRIFT, and 1H and 13C-NMR spectroscopy. Fresh plants were subsequently incubated for 6 months in an artificial mineral soil, and the HA-like and core-HA-like trends were monitored quantitatively (C fraction content) and qualitatively (spectroscopic approach) in order to study their contribution to the formation of soil humic acid. During incubation the HAC like partially degraded (loss of 320 gkgHAC(-1)) and partially formed new fulvic like acids (160 gkgHAC(-1)). On the contrary, the stable fraction of HAC, the core-HAC-like, was maintained (loss of 153 kgcore-HAC(-1)), representing, after incubation, 846 gkg(-1) of the initial core-HAC-like content. The core-HA-like fraction is composed of lignin residues, polysaccharides, lipids and proteins, probably structured into a well-defined network, i.e. the plant cell wall. PMID- 15109876 TI - Fungal laccase-catalyzed degradation of hydroxy polychlorinated biphenyls. AB - Hydroxy polychlorinated biphenyls (hydroxy PCBs) are toxic metabolites of PCBs. Their toxicity such as strong endocrine disruption demands effective remediation methods. Laccases from Trametes versicolor and Pleurotus ostreatus were tested to degrade hydroxy PCBs. Optimum pHs for both enzymes were around 4.0. Laccase from T. versicolor degrades hydroxy PCBs more rapidly than that from P. ostreatus. The enzymatic activities remained little changes in up to 10% organic solvents, but decreased rapidly in more than 10% acetone, acetonitrile or DMSO. Degradation rate constants decreased with increase of chlorination and no degradation was observed with tetra-, penta- and hexa-chloro hydroxy PCBs in non-mediated reactions. However, the tetra- to hexa-chloro hydroxy PCBs were degraded by laccase from T. versicolor in the presence of the mediator 2,2,6,6 tetramethylpiperidine-N-oxy radical. The other mediators, 4-ethyl-2 methoxyphenol, 2,2'-azino-bis(3-ethylbenzthiazoline sulfonic acid) diammonium salt and 1-hydroxybenzotriazole and humic acid, also enhanced degradation of all the hydroxy PCBs except 4-hydroxy-2',3,3',4',5,5'-hexachlorobiphenyl. The results showed that 3-hydroxy biphenyl was more resistant to laccase degradation than 2- or 4-hydroxy analogues. Significant linear-correlations (coefficient of determination, r2 = 0.9097 and 0.8186 for laccases from P. ostreatus and T. versicolor, respectively) were found between the ionization potentials and the removal rate constants of hydroxy PCBs. PMID- 15109877 TI - Effect of dye compounds on the adsorption of atrazine by natural sediment. AB - The overall objective of this research is to investigate competitive adsorption between atrazine (AT) and dye compounds in the natural aquatic sediment. The sorbent was sediment obtained from Guanting Reservoir (Beijing, China), which contained 25% sand, 67% silt, 8% clay, and 2.06% organic carbon. Batch adsorption experiments were conducted at various Ca2+ concentration, pH levels, temperatures, and introducing conditions of dye compounds. Compared with the dye free system, both of dyes including Congo red (CR) and methylene blue (MB) reduce the adsorption of atrazine over the range of dye concentrations examined, with the adsorption percentage of atrazine decreasing about 14-30%. And the competition between AT and MB is much stronger than that between AT and CR. The adsorption experimental data points have been fitted to the Freundlich equation in order to calculate the adsorption capacities (Kf) of the samples; Kf values range from 1.669 micromol/kg for the MB-AT sample up to 3.738 micromol/kg for the AT-alone sample. By contrast with the single-solute adsorption isotherm, both simultaneous adsorption and dye preloading inhibit the adsorption of atrazine. As for AT preloading, the impacts of CR and MB are different on the desorption of atrazine. As compared to the atrazine desorption without dye compounds, a certain amounts of atrazine molecules are replaced by MB in AT preloading system, while in CR solution AT is adsorbed strongly on the sediment and could not be replaced by CR. The result suggests that micropore constriction by CR reduces the desorption rate of atrazine. PMID- 15109878 TI - Ion mobility based on column leaching of South African gold tailings dam with chemometric evaluation. AB - New column leaching experiments were designed and used as an alternative rapid screening approach to element mobility assessment. In these experiments, field moist material was treated with an extracting solution to assess the effects of acidification on element mobility in mine tailings. The main advantage of this version of column leaching experiments with partitioned segments is that they give quick information on current element mobility in conditions closely simulating field conditions to compare with common unrepresentative air-dried, sieved samples used for column leaching experiments. Layers from the tailings dump material were sampled and packed into columns. The design of columns allows extracting leachates from each layer. The extracting solutions used were natural (pH 6.8) and acidified (pH 4.2) rainwater. Metals and anions were determined in the leachates. The concentrations of metals (Ca, Mg, Fe, Mn, Al, Cr, Ni, Co, Zn, and Cu) in sample leachates were determined using ICP OES. The most important anions (NO3-, Cl-, and SO4(2)-) were determined using the closed system izotacophoresis ITP analyser. The chemical analytical data from tailings leaching and physico-chemical data from field measurements (including pH, conductivity, redox potential, temperature) were used for chemometric evaluation of element mobility. Principal factor analysis (PFA) was used to evaluate ions mobility from different layers of tailings dump arising from varied pH and redox conditions. It was found that the results from the partitioned column leaching illustrate much better complex processes of metals mobility from tailings dump than the total column. The chemometric data analysis (PFA) proofed the differences in the various layers leachability that are arising from physico-chemical processes due to chemical composition of tailings dump deposit. PMID- 15109879 TI - Variability of parameters measured during the resuspension of sediments with a particle entrainment simulator. AB - The release of contaminants from sediments is an important problem facing environmental managers concerned with issues such as maintenance dredging, habitat restoration and dredge spoil placement. While there are laboratory methods to assess the remobilization of contaminants from resuspended sediments, little is known about how their operating characteristics influence a sediments response to resuspension. In this study, a particle entrainment simulator (PES) for resuspending sediments was evaluated by assessing variability among replicates and over operation time (12 h) for three estuarine sediments. These sediments varied in physical and chemical properties as well as the degree of metal contamination. Results showed that under identical resuspension conditions, there was low variability among sediment replicates. Sediment properties and the degree of metal contamination affected the magnitude and variation in water column parameters and contaminant release. Fine-grained sediments affected water column parameters, which stabilized over several hours of resuspension. While metal concentrations in the aqueous and particulate samples varied throughout operation, variation was highest for Cd and Ni. Compared to other methods used to evaluate the effects of sediment resuspension, the PES has operational advantages such as the ability to monitor water column conditions and resuspend sediment at specified energy levels. Overall, the PES provided reproducible conditions of resuspension effects among sediments varying in composition and contamination, demonstrating its value as a tool to evaluate contaminant mobility. PMID- 15109880 TI - The utilization of the Antarctic environmental specimen bank (BCAA) in monitoring Cd and Hg in an Antarctic coastal area in Terra Nova Bay (Ross Sea--Northern Victoria Land). AB - The first projects relating to levels of Cd and Hg on marine biota and sediments from Terra Nova Bay (Ross Sea--Antarctica) and their bioaccumulation and biomagnification in this trophic web have been carried out by research programmes pertaining to the Italian Antarctic Research Program (PNRA) since 1989. Making use of this data, and checking the same metals after 10 years thanks to the samples stored in the BCAA, we have looked for the levels of Cd and Hg in a coastal marine ecosystem of Terra Nova Bay, and have proposed using some organisms to monitor the levels of these two heavy metals in this environment where the Italian Base is located, using the data determinate in this work as background levels. In our work, the amount of Hg and Cd concentrations have been determined in biota from the inner shelf of Terra Nova Bay (Adamussium colbecki, Laternula elliptica, Odontaster validus, Sterechinus neumayeri, Trematomus bernacchii, Iridaea cordata, Phyllophora antarctica, Parborlasia corrugatus), and in two different size fractions of sieved marine sediments (<2000 microm and <63 microm). To widen the distribution of Cd and Hg in this ecosystem we have also investigated the fraction of these metals bound to the labile phase of the marine sediments, and their presence in the particulate matter found in pack-ice cores, recent snow, water column and sea microlayer. PMID- 15109881 TI - Determination of volatile organic compounds in workplace air by multisorbent adsorption/thermal desorption-GC/MS. AB - Investigation of volatile organic compounds (VOCs) was first conducted in the air of class-100 cleanrooms at liquid crystal display (LCD) fabrication facilities. Air samples were collected on multisorbent tubes (including Carbopack B, Carbopack C, and Carbosieve S-III) and analyzed using adsorption/thermal desorption coupled with gas chromatography-mass spectrometry (GC-MS). Optimal conditions lead to average recoveries in the range of 96.2-98.2%, and method detection limits between 0.38 and 0.78 ppb, under the condition of 1-l sampling volume and 80% relative humidity. The method appears to be accurate, sensitive, simple and well-suited for determining VOC distributions from various stages of LCD manufacturing process and temporal variations of the analyte concentrations. About 15 VOCs were identified in workplace air. The major pollutants such as propylene glycol methyl ether acetate (PGMEA), butyl acetate, and acetone that are commonly used in the opto-electronics industry were detected and accurately quantified with the established method. PMID- 15109882 TI - An evaluation of two fluorescence screening methods for the determination of chrysene metabolites in fish bile. AB - Two screening methods, synchronous fluorescence spectrometry (SFS) and high performance liquid chromatography with fluorescence detection (HPLC-F), have been evaluated for their suitability in determining chrysene metabolites in fish bile. The optimal wavelength pair, excitation/emission 272/374 nm, for SFS measurements of chrysene metabolites was identified by analysis of bile taken from fish exposed to the pure compound. This analysis revealed in addition some information about the metabolite pattern. However, when bile from fish exposed to complex and environmentally relevant mixtures of polycyclic aromatic hydrocarbons (PAHs) was analysed using these methods, identification of the chrysene metabolites was poor. Analysis of bile taken from fish exposed to single PAHs identified other three- and four-ring aromatics as the main interfering compounds. Both methods were equally able to discriminate between impacted and reference sites by determination of relative concentrations of fluorescent three- and four-ring aromatics. PMID- 15109883 TI - Nicotine, its metabolism and an overview of its biological effects. AB - Nicotine is a naturally occurring alkaloid found in many plants. The principal sources of nicotine exposure is through the use of tobacco, nicotine containing gum and nicotine replacement therapies. Nicotine is an amine composed of pyridine and pyrrolidine rings. It has been shown that nicotine crosses biological membranes and the blood brain barrier easily. The absorbed nicotine is extensively metabolized in the liver to form a wide variety of metabolites including nicotine N'-oxide and cotinine N'-oxide. These are the products of mixed function oxidase system. Nicotine is also converted to some biologically important compounds during harvesting. Among these are the nitrosamines specific to tobacco. Nicotine has been shown to affect a wide variety of biological functions ranging from gene expression, regulation of hormone secretion and enzyme activities. The objective of this study was to overview the biological effects and metabolism of nicotine. PMID- 15109884 TI - Pharmacological evidence for a presynaptic action of venoms from Bothrops insularis (jararaca ilhoa) and Bothrops neuwiedi (jararaca pintada). AB - Whereas the presynaptic action of Crotalus durissus terrificus venom is well established, Bothrops venoms have historically been considered to have only postsynaptic and muscular effects. However, some studies have also suggested a presynaptic action for these venoms. In this work, we used chick biventer cervicis preparations to compare the presynaptic actions of two Bothrops venoms (B. insularis and B. neuwiedi) with that of C. d. terrificus venom. At 10 microg/ml, all venoms produced irreversible blockade of the twitch tension responses, with no reduction in acetylcholine (ACh)-induced contractures and only a slight decrease in potassium induced-contractures. The times (in min) required to produce 50% neuromuscular blockade (C. d. terrificus: 16.3+/-0.7, n = 8; B. insularis: 30.0+/-1.9, n = 5; B. neuwiedi: 42.0+/-2.0, n = 8; mean +/- SEM) were significantly different among the venoms (p < 0.01). Lowering the temperature at which the experiments were done (from 37 to 24 degrees C) prevented neuromuscular blockade by the three venoms, indicating that enzyme activity may be involved in this response. At concentrations capable of causing complete neuromuscular blockade, creatine kinase release remained close to levels seen in control preparations incubated with Krebs solution alone (500-1200 IU/l). Commercial crotalic antivenom, but not bothropic antivenom, protected against the neuromuscular blockade caused by B. insularis and B. neuwiedi venoms. These observations indicate that bothropic venoms may contain components which act presynaptically in a manner similar to C. d. terrificus venom, and that at low venom concentrations a direct action on skeletal muscle does not contribute to this presynaptic neurotoxicity. PMID- 15109885 TI - Occurrence and elimination of cyanobacterial toxins in two Australian drinking water treatment plants. AB - In Australian freshwaters, Anabaena circinalis, Microcystis spp. and Cylindrospermopsis raciborskii are the dominant toxic cyanobacteria. Many of these surface waters are used as drinking water resources. Therefore, the National Health and Medical Research Council of Australia set a guideline for MC LR toxicity equivalents of 1.3 microg/l drinking water. However, due to lack of adequate data, no guideline values for paralytic shellfish poisons (PSPs) (e.g. saxitoxins) or cylindrospermopsin (CYN) have been set. In this spot check, the concentration of microcystins (MCs), PSPs and CYN were determined by ADDA-ELISA, cPPA, HPLC-DAD and/or HPLC-MS/MS, respectively, in two water treatment plants in Queensland/Australia and compared to phytoplankton data collected by Queensland Health, Brisbane. Depending on the predominant cyanobacterial species in a bloom, concentrations of up to 8.0, 17.0 and 1.3 microg/l were found for MCs, PSPs and CYN, respectively. However, only traces (<1.0 microg/l) of these toxins were detected in final water (final product of the drinking water treatment plant) and tap water (household sample). Despite the low concentrations of toxins detected in drinking water, a further reduction of cyanobacterial toxins is recommended to guarantee public safety. PMID- 15109886 TI - Proteomics approach on microcystin binding proteins in mouse liver for investigation of microcystin toxicity. AB - Microcystins (MC) produced by freshwater cyanobacteria are potent hepatotoxins. MC inhibit protein phosphatases (PP) 1 and 2A. MC and okadaic acid (OA), which is a similar PP inhibitor whereas it has a less affinity to PP1 than PP2A, behave similarly to primary culture hepatocytes, with inducements of phosphorylations of cytoskeleton, morphological changes and apoptosis. Although the distribution of OA in mouse liver was observed immunohistochemically, no OA injury was found. The purpose of this study was therefore to determine why only MC has specific toxicities on the liver. A systematic process of MC affinity chromatography and proteomics, using two-dimensional gel electrophoresis and MALDI-TOFMS, indicated the existence of some MC-binding proteins including the complexes of PP1, PP2A, and PP4 with their own regulatory subunits in mouse liver extracts. The competitive inhibition experiments using affinity chromatography with OA showed that two of the three protein complexes strongly interacted with OA, whereas only the complex of PP1 with the inhibitory subunit NIPP1 did not strongly interacted with OA. These results suggest that the PP1 complex is not related to the common behavior of MC and OA of primary culture hepatocytes, and is related to the specific hepatotoxicities of MC. PMID- 15109887 TI - Quantitative variability in the biodistribution and in toxinokinetic studies of the three main alpha toxins from the Androctonus australis hector scorpion venom. AB - Scorpion stings represent a medical problem in numerous countries. The scorpion Androctonus australis hector produces three alpha toxins (Aah I to III), which are responsible for most of the lethality in mammals. These toxins act on sodium channel and do not cross-react immunologically. We used RIA and ELISA to measure the concentrations of these three toxins in plasma, urine and different organs after i.v. and s.c. injections of water extracts of venoms in rabbits or mice. In both animals, the toxins rapidly appeared in plasma after s.c. injection as it was previously described for the whole venom. However, the toxins disappeared from the blood more quickly than did other main components of the venom. Thus, serotherapy must be initiated immediately to prevent the toxin from reaching its target. We also detected the toxins in urine, kidneys, heart and lungs, but not in the brain. However, the concentration of Aah II was always lower than that of Aah I. Analysis of five samples of venom collected in different areas of southern Tunisia showed that a large polymorphism exists for the three toxins. This is yet another difficulty for serotherapy as there is no cross-antigenicity between them. PMID- 15109888 TI - Isolation, molecular cloning and functional characterization of a novel beta toxin from the Venezuelan scorpion, Tityus zulianus. AB - Sting in children by Tityus zulianus scorpions (western Venezuela) often produces cardiorespiratory arrest and death by pulmonary oedema. To assess its toxicity, lethality in mice of T. zulianus soluble venom was determined. Toxin composition was studied by fractionating the crude venom through reversed-phase HPLC. The most abundant peptide, Tz1, was purified further and its N-terminal sequence, amino acid composition and molecular mass (by electron-spray ionization mass spectrometry) determined. In the presence of Tz1, activation of recombinant rat skeletal muscle sodium channels (Na(V)1.4) was shifted about 35 mV in the hyperpolarizing direction in a prepulse-dependent manner. This typical beta-toxin effect had an apparent EC50 of 3.5 microM A cDNA sequence encoding Tz1 was isolated from T. zulianus venom gland RNA using a combination of 5'- and 3'-RACE PCR. Analysis of the encoded sequence indicated that Tz1 is the processed product of a precursor containing: (i) a 20-residue long leader peptide; (ii) the amino acid sequence of the mature toxin (64 residues); and (iii) an extra Gly-Lys tail at the C-terminus, probably removed post-translationally. A comparison of Tz1 with Tityus serrulatus beta-toxin Ts1 revealed that some of the non-conservative replacements in Tz1 lie in regions potentially involved in receptor recognition. PMID- 15109889 TI - The in vivo cardiovascular effects of box jellyfish Chironex fleckeri venom in rats: efficacy of pre-treatment with antivenom, verapamil and magnesium sulphate. AB - Using a new technique to extract venom from the nematocysts, the efficacy of CSL box jellyfish antivenom (AV) and adjunct therapies, verapamil and magnesium sulfate (MgSO(4)), were investigated against the in vivo cardiovascular effects of Chironex fleckeri venom in anaesthetised rats. C. fleckeri venom (30 microg/kg; i.v.) produced a transient hypertensive response followed by hypotension and cardiovascular collapse within 4 min of administration. Prophylactic treatment of anaesthetised rats with CSL box jellyfish AV (3000 U/kg; i.v.) did not have any effect on the venom-induced pressor response, but prevented cardiovascular collapse in four out of 10 animals. Administration of verapamil (20mM@0.25 ml/min; i.v.) either alone or in combination with AV, did not have any effect on the C. fleckeri venom-induced pressor response nor the consequent hypotension or cardiovascular collapse of animals. However, the administration of verapamil negated the partially protective effects of AV. Concurrent artificial respiration of animals with the above treatments did not attenuate the C. fleckeri venom-induced cardiovascular effects. MgSO(4) (0.05 0.07M@0.25 ml/min; i.v.) alone did not have any effect on the venom-induced pressor response nor the consequent cardiovascular collapse of animals. However, although combined AV and MgSO(4) administration could not inhibit the transient pressor effect following the administration of C. fleckeri venom, it prevented cardiovascular collapse in all animals. We show for the first time, the cardiovascular effects of a C. fleckeri venom sample free of tentacular contamination and the potential of MgSO(4) as an adjunct therapy for the treatment of potentially fatal C. fleckeri envenomings. PMID- 15109890 TI - Vipera palaestinae envenomation in 327 dogs: a retrospective cohort study and analysis of risk factors for mortality. AB - Vipera palaestinae (Vp), formerly a subspecies of the near east viper Vipera xanthina, is the most common poisonous snake in Israel and neighbouring countries (Jordan, Lebanon and Syria), and is responsible for most envenomations in humans and domestic animals. Hospital records were retrospectively reviewed for confirmed cases of Vp envenomations in dogs over a 13-year period and 327 cases were included in the study. Most envenomations occurred between May and October, and between 02:00 and 10:00 PM. The most frequent clinical signs included: local swelling and oedema (99.6%), viper teeth penetration marks (51%), tachypnoea (50%), panting (44%), increased body temperature (19.2%), tachycardia (>160/min, 19%), salivation (18%) and lameness (15.6%). Common haematological findings included: increased haematocrit (47%), increased haemoglobin concentration (45%), leucocytosis (39%), and thrombocytopenia (30%). The prothrombin time and activated partial thromboplastin time were prolonged in 68 and 21% of the dogs, respectively. Blood biochemistry abnormalities included increased activities of muscle enzymes, hyperglycaemia, hyperbilirubinaemia, hyperglobulinaemia and hypocholesterolaemia. The mortality rate was 4% (13 dogs). The following variables were significantly (p < 0.05) associated with mortality: body weight below 15 kg (p = 0.01), limb envenomation (0.008), envenomation at night (p = 0.025), severe lethargy (P < 0.001), hypothermia (p = 0.04), systemic bleeding (p = 0.001), shock (p = 0.007), dyspnoea (p = 0.002), tachycardia (p = 0.002), thrombocytopenia (p = 0.02), and glucocorticosteroid therapy (p = 0.002). Dogs younger than 4 years had a lower death risk (p = 0.01). The association of steroid therapy with increased mortality suggests that the use of steroids in Vp envenomations may be harmful. Specific antivenom therapy (10 ml/dog) was not associated with a higher survival rate, thus its use, dose and timing of administration should be further investigated. PMID- 15109891 TI - Confirmation of brevetoxin metabolism in cockle, Austrovenus stutchburyi, and greenshell mussel, Perna canaliculus, associated with New Zealand neurotoxic shellfish poisoning, by controlled exposure to Karenia brevis culture. AB - We examined metabolism of PbTxs in New Zealand cockle, Austrovenus (A.) stutchburyi, and greenshell mussel, Perna (P.) canaliculus, by means of liquid chromatography coupled with tandem mass spectrometry. PbTx-2, PbTx-3 and BTX-B5 were detected in Karenia (K.) brevis culture medium in the ratio of ca. 50:2:5. The amounts of PbTx-3 and BTX-B5 were greatly increased in both seawater and shellfish exposed to K. brevis cultures or supernatant prepared by disruption of K. brevis under appropriate condition, while those of PbTx-2 were decreased. Some PbTx-2 was present in P. canaliculus, but not in A. stutchburyi. Low levels of BTX-B1 were detected in A. stutchburyi, but not P. canaliculus. Levels of PbTx-3 and BTX-B5 were highest immediately after exposure and then declined rapidly in both shellfish. BTX-B1 increased in concentration after exposure, and was then gradually eliminated from A. stutchburyi. Three successive exposures of A. stutchburyi to K. brevis cultures resulted in similar initial levels of PbTx-3 and BTX-B5, while BTX-B1 accumulated after each dose. In P. canaliculus, initial levels of PbTx-3 were similar, while PbTx-2 and BTX-B5 accumulated after each dose. PbTx-3 and BTX-B5 are proposed to be suitable markers for monitoring shellfish toxicity after a red tide event. PMID- 15109892 TI - Comparison of paralytic shellfish poisoning toxin between carnivorous crabs (Telmessus acutidens and Charybdis japonica) and their prey mussel (Mytilus galloprovincialis) in an inshore food chain. AB - Paralytic shellfish poisoning toxin in two shore crab species, Telmessus acutidens and Charybdis japonica, were compared with the toxin in the prey mussel Mytilus galloprovincialis and causative dinoflagellates Alexandrium tamarense, all having been collected at Onahama, Fukushima Prefecture, in the northern part of Japan. When the toxicities were detected in mussels by mouse bioassays, 73.7% of the sampled T. acutidens were toxic in the hepatopancreas. T. acutidens has been found to become toxic for three years, therefore, it can be concluded that the crab commonly and repeatedly accumulate the toxins via the food chain at Onahama. C. japonica was also expected to be a possible vector species, because small quantities of the toxins were detected in eight specimens of the crab by HPLC analysis. By the comparison of the toxin profiles in the dinoflagellates, mussels and the crab T. acutidens, reductive conversions of GTX1 and GTX4 were observed when the toxins passed through the three species in the food chain. But increases of STX and neoSTX by further reductive process were not observed in the crab. The absence of the STX group toxins in the crab suggests that the crab eliminates the toxin before such reductive process occur. PMID- 15109893 TI - Channel-forming activity in the venom of the cockroach-hunting wasp, Ampulex compressa. AB - The parasitoid solitary wasp Ampulex compressa uses the cockroach Periplaneta americana as a food supply for its larvae. To subdue its prey, the wasp injects a venom cocktail into the brain of the cockroach. We investigated channel activity of A. compressa venom by collecting venom and incorporating it into a planar lipid bilayer. The venom, reconstituted into the bilayer, showed ion channel activity, forming a fast-fluctuating channel with a small conductance of 20+/ 0.1pS, with no voltage sensitivity. These channels were not observed when the venom was digested with proteases before application to the bilayer, but were not affected by exposure to protease after their incorporation into the bilayer, indicating that the active venom component is a peptide. The channels were found to be cation selective with similar selectivity for the monovalent cations K(+), Li(+) and Na(+), but showed high selectivity against anions (Cl(-)) and divalent cations (Ca(2+) and Mg(2+)). This study is the first demonstration and biophysical characterization of channel activity in the venom of A. compressa. The possible functional significance of this channel activity is discussed in light of the unusual nature of the effects of this wasp venom on the behavior of its prey. PMID- 15109894 TI - Ability of eugenol to reduce tongue edema induced by Dieffenbachia picta Schott in mice. AB - Dieffenbachia picta Schott (Araceae), known in Brazil as "comigo-ninguem-pode" is an ornamental plant with toxic properties. Its juice, when chewed, causes a painful edema of the oral mucous membranes, buccal ulcerations and tongue hypertrophy. This acute inflammation sometimes becomes severe enough to produce glottis obstruction, respiratory compromise and death. Eugenol (4-alil-2 metoxiphenol), the essential oil extracted from Caryophyllus aromaticus (Myrtaceae) is widely used in odontology. In this study, our objective was to standardize, in mice, a measurable methodology for the tongue edema induced by the topical application of the D. picta stem juice; evaluate the effects of eugenol in this model and compare the results with emergency treatment used in hospitals. Our results show that in spite of a small increase in edema a few minutes after administration, emergency treatment reduced by 70% the overall edema. When compared with the combination of the above drugs, eugenol, even at the smallest dose of 5 microg/kg, regardless of the chosen administration route, or the moment the treatment began, presents better results in the reduction and inhibition of the tongue edema induced by the D. picta juice. PMID- 15109895 TI - Amino acid sequence and function of a new alpha-toxin from the Amazonian scorpion Tityus cambridgei. AB - A toxic peptide earlier denominated Tc48b [Toxicon 40 (2002) 557] was purified to homogeneity and its amino acid sequence determined. It has 64 amino acid residues stabilized by four disulfide bridges with a molecular weight of 7,385.2 atomic mass units (a.m.u.). It affects Na(+)-permeability in pituitary GH3 cells in culture, in a similar fashion as those reported for alpha-scorpion toxins, contrary to most of the New World scorpion toxins that are beta-toxins. PMID- 15109896 TI - Sources of spores for the possible horizontal transmission of Thelohania solenopsae (Microspora: Thelohaniidae) in the red imported fire ants, Solenopsis invicta. AB - We screened adult and larval secretions and midden piles for the presence of Thelohania solenopsae spores to decipher potential sources for the horizontal transmission of the pathogen in fire ants. Hemolymph samples from both adult and larvae were also screened to rule out hemolymph contamination of samples. In adults, Thelohania spores were found in the crop and the fecal fluids, although only free spores were found in the fecal fluids of adults. In fourth instar larvae, both free and octospores were seen in midgut and the meconium samples. All of the midden pile samples had T. solenopsae spores of both types. Based on these results, we theorize that the pathogen may be horizontally transmitted within a colony by the removal and sharing of meconium of prepupating fourth instar larvae by adult workers and by the adult fecal droppings, and intercolonially by contamination of midden piles or brood raiding. PMID- 15109897 TI - Pathogenicity and virulence. AB - Invertebrate pathologists have multiple definitions for the terms pathogenicity and virulence, and these definitions vary across disciplines that focus on host pathogen interactions. We surveyed various literatures, including plant pathology, invertebrate pathology, evolutionary biology, and medicine, and found most define pathogenicity as the broader term, which incorporates virulence. Virulence is seen as the severity of disease manifestation that can only be measured in infected individuals. These definitions readily apply to both lethal and non-lethal diseases. Invertebrate pathologists commonly use dose-response bioassays to estimate LD(50) or LC(50) (dose or concentration needed to kill 50% of hosts exposed). These bioassays measure pathogenicity if the bioassay includes a transmission component, and measure virulence if the bioassay is measured in infected individuals only. Another common bioassay estimate is LT(50) (median time to death of infected hosts), which is a measure of virulence as long as survivors are not included in its calculation. PMID- 15109898 TI - Comparative analysis of the production of insecticidal and melanizing macromolecules by strains of Beauveria spp.: in vivo studies. AB - Eleven strains of Beauveria bassiana, and a further five species of Beauveria sp., were tested by injection of 8x10(2) conidia into the haemocoel of the larvae of the lepidopteran Galleria mellonella with the aim of analysing their toxin producing activity in vivo. Although the virulent strains killed 100% of the insects at slightly different rates (4-6 days) there were significant differences in the pattern and intensity of host melanization caused by isolates. The majority of the isolates of Beauveria spp. induced a fast and intense melanization of the cuticle of the integument and of tracheal wall, which followed one of three patterns. Another small group of two B. bassiana strains, isolated from Ostrinia nubilalis, induced very weak or no melanization. Strains 618 and 101 of B. bassiana, were selected as models of "melanizing" and "non melanizing" strains, respectively. Ultrastructural alterations of cells of hypodermal and tracheal epithelium and of haemocytes, assumed to be at least partially caused by fungal toxins, were revealed in larvae infected by both isolates. However, their effects on the fine structure of the hypodermis were different. Injection of sera obtained from haemolymph of insects infected with B. bassiana 618 showed that they have insecticidal, melanizing, and cytotoxic effects similar to those occurring during mycosis. Chromatographic studies and bioassays with fractions prepared from crude serum have allowed a partial identification of the toxic molecules secreted by the fungus in vivo. They are proteinaceous, as shown by protease treatments, thermolabile, negatively charged, and not glycosylated with alpha-d-mannose or alpha-d-glucose. If strain B. bassiana 618 produces melanizing macromolecules which are vivotoxins secreted during the mycosis, the mode of action of isolate 101 is different. Its capacity to kill the host depends on active mycelial development, and on the production of low molecular weight toxins. PMID- 15109899 TI - Investigations on the destruxin production of the entomopathogenic fungus Metarhizium anisopliae. AB - The dynamics of cyclic peptide destruxins (dtxs) produced by Metarhizium anisopliae strains V245 and V275 were monitored both on solid and in liquid media. The results showed that both strains did not produce dtxs in large-scale fermenter cultures or solid Czapek Dox (CD) agar. Production of the major dtxs A and B could be determined in both strains when grown on rice for up to 10-30 days. The main dtxs A, B, E, and E diol were detected in CD liquid culture filtrate from both strains after three days post-inoculation on. Parallel decrease of dtx E and increase of E diol in the culture medium were found, indicating that the latter is the hydrolytic product from the former. Production of dtxs A and B was significantly positively correlated. A negative correlation was observed between the production of the metabolites and pH value of the medium. The influence of different nutrient sources on dtx production was evaluated by using media with different carbon and nitrogen ratios as well as with different insect homogenates. The findings showed that the amount of dtxs A, B, and E increased with the increasing content of peptone in the medium. When insect homogenate was used as single nutrient source or added to CD medium, no toxins were detected in the culture filtrate. The potential risk posed by the toxic metabolites during mass production is discussed. PMID- 15109900 TI - Effects of host desiccation on development, survival, and infectivity of entomopathogenic nematode Steinernema carpocapsae. AB - This study investigates the effect of host desiccation on entomopathogenic nematode (EPN) development, emergence, infectivity, and cross-protection against secondary environmental stress. Galleria mellonella hosts infected with the EPN Steinernema carpocapsae A10 were allowed to dehydrate in an environmental chamber for up to 56 days at 23 degrees C achieving a weight loss of approximately 86% by day 44 post-infection. Host carcasses were rehydrated on water-saturated filter paper in White traps to collect emergent infective juveniles (IJ) at specific time intervals. Populations were counted with an apparent peak coinciding with desiccated hosts rehydrated at 24-day post-infection. Desiccation-stressed IJ populations from each time interval were tested for infectivity, and cross resistance to secondary temperature and pH stresses and were found to have significant increases in both infectivity and protection from extremes of temperature and pH compared with controls. Total aqueous soluble protein profiles from control and desiccation-stressed IJs were analyzed using 10% SDS Laemmli gels. Several novel proteins were over-expressed in EPN from hosts subjected to desiccation suggesting the induction and expression of stress response genes. PMID- 15109901 TI - Lyophilization of lepidopteran midguts: a preserving method for Bacillus thuringiensis toxin binding studies. AB - Binding assays with brush border membrane vesicles (BBMV) from insect midguts are commonly used in the study of the interactions between Bacillus thuringiensis Cry toxins and their receptors. Collaboration between laboratories often require that frozen insect samples are sent in dry ice. Because of customs restrictions and delays, sample thawing is always a risk and often the biological material becomes ruined during shipping. We have tested lyophilization as an alternative method for preserving insect midguts for binding studies with B. thuringiensis Cry toxins. For this purpose, BBMV were prepared from both frozen and lyophilized midguts from three lepidopteran species: Spodoptera exigua, Manduca sexta, and Helicoverpa armigera. Higher membrane protein recovery was always obtained from lyophilized midguts compared to frozen midguts, and similar membrane marker enzyme activities were found in BBMV from either treatment. Comparable equilibrium dissociation constants and binding site concentrations, calculated from binding experiments with labeled (125)I-Cry1Ab toxin, were found using BBMV from either method. In the light of these results, lyophilization is a good preserving method of lepidopteran midguts to study binding of B. thuringiensis Cry toxins. PMID- 15109902 TI - Seasonality and prevalence rates of microsporidia in Simulium pertinax (Diptera: Simuliidae) larvae in the region of Serra dos Orgaos, Rio de Janeiro, Brasil. PMID- 15109903 TI - Effects of testosterone on the development of neuromuscular systems and their target tissues involved in courtship and copulation in green anoles (Anolis carolinensis). AB - Male green anole lizards court females using a red throat fan (dewlap) and copulate by intromitting one of two penises (hemipenes). These structures begin sexually monomorphic, but by adulthood males have larger dewlaps, only males have hemipenes, and many of the neuromuscular components of both systems show male biased dimorphisms. We hypothesized that testosterone (T), which increases in juvenile males but not females about a month after hatching, facilitates masculinization. To test this idea, on post-hatching day 30, gonadally intact females received either a blank or T implant, and males were either castrated or sham-castrated. At day 90, juveniles were euthanized and the length of the cartilage and cross-sectional areas of the muscle fibers and motoneurons required for dewlap extension were examined. We also measured the cross-sectional areas of the hemipenes and associated muscle fibers and motoneurons, and counted the motoneurons. T-treated females had longer cartilages and larger dewlap muscle fibers compared to those with blank implants. No effects on motoneurons were detected, and no females possessed hemipenes or associated musculature. In males, castration produced shorter dewlap cartilages and smaller hemipenes; other measures were not affected by treatment. These data indicate that components of the dewlap system differentiate relatively late in development, that T likely mediates the process, and that although components of the copulatory system are plastic in juvenile males, sexual differentiation of peripheral features is complete before day 30. The data also suggest that target structures (dewlap cartilage and hemipenes), compared to their neuromuscular effectors, are particularly sensitive to developmental T exposure. PMID- 15109904 TI - Effects of 17-beta-estradiol treatment of female zebra finches on offspring sex ratio and survival. AB - Treatment of female zebra finches (Taeniopygia guttata) with 17-beta-estradiol leads to a female-biased sex ratio in their offspring at the age of independence [Horm. Behav. 35 (1999) 135]. It is unclear whether this is due to a bias of the primary sex ratio or to sex-specific survival. We replicated this experiment and found again a significantly higher total number of daughters than sons at independence in the estradiol-treated group. This was due to higher embryonic survival of daughters compared with sons in the estradiol-treated group and the reverse in the control group. There was no effect of the hormone treatment on the primary sex ratio. Treatment with 17-beta-estradiol led to a significantly shorter hatching time and to heavier offspring at day 7 after hatching. This weight was correlated with maternal plasma estradiol levels on the day of the first egg, which were significantly higher in the estradiol-treated group than in the control group. The results do not support the idea that maternal estradiol levels influence the primary sex ratio. They indicate that maternal estradiol differentially affects survival of sons and daughters via an influence on the embryonic environment, possibly enhancing offspring growth. PMID- 15109905 TI - Hormonal status and test condition, but not sexual experience, modulate partner preference in female rats. AB - A series of experiments was conducted to determine the contributions of hormonal status, test condition, and sexual experience to the display of partner preference by female rats. Preference for a sexually active male rat over a sexually receptive female rat was assessed in independent groups of female rats tested in a condition limiting physical contact (No Contact) and a condition allowing for sexual interaction (Contact). Although hormonal status and test condition influenced the preference for a sexually active male, repeated testing and sexual experience had no effect. Experiment 1 demonstrated that independent of test condition, preference for the male is stronger in estrogen- and progesterone-primed rats than in rats receiving the vehicle. Moreover, independent of hormone condition, rats tested in the No Contact condition exhibit a stronger preference for the male than rats tested in the Contact condition, reflecting in part the active pacing of mating stimulation by sexually receptive rats tested in the Contact condition. Experiment 2 showed that the overall pattern of partner preference in proestrous and diestrous rats was similar to that observed in ovariectomized, estrogen- and progesterone-primed, and oil treated rats, respectively. In Experiment 3, rats primed with estrogen alone did not exhibit a preference for the male even though fully receptive. Experiments 4 and 5 demonstrated that sexual experience does not affect the expression of preference for the male in estrogen- and progesterone-primed rats. The present findings demonstrate that the female rat's preference for the male is stable across repeated tests and is not affected by sexual experience. Our results also confirm that gonadal hormones influence the expression of a preference for a sexually active male versus a sexually receptive female and demonstrate that the magnitude of preference is modulated by test conditions. PMID- 15109906 TI - Behavioral and neuroendocrine correlates of displaced aggression in trout. AB - In humans and other primates, violent actions performed by victims of aggression are often directed toward an individual or object that is not the source of provocation. This psychological phenomenon is often called displaced aggression. We demonstrate that displaced aggression is either rooted in evolutionarily conserved behavioral and neuroendocrine mechanisms, or represent a convergent pattern that has arisen independently in fish and mammals. Rainbow trout that briefly encountered large, aggressive fish reacted with increased aggression toward smaller individuals. There was a strong negative correlation between received aggression and behavioral change: Individuals subjected to intense aggression were subdued, while moderate assaults induced strong agitation. Patterns of forebrain serotonin turnover and plasma cortisol suggest that the presence of socially subordinate fish had an inhibitory effect on neuroendocrine stress responses. Thus, subordinate individuals may serve as stress-reducing means of aggressive outlet, and displaced aggression toward such individuals appears to be a behavioral stress coping strategy in fishes. PMID- 15109907 TI - Shifts in preferred learning strategy across the estrous cycle in female rats. AB - The current status of the effects of ovarian steroids on learning and memory remains somewhat unclear, despite a large undertaking to evaluate these effects. What is emerging from this literature is that estrogen, and perhaps progesterone, influences learning and memory, but does so in a task-dependent manner. Previously, we have shown that ovariectomized rats given acute treatments of estrogen acquire allocentric or "place" tasks more easily than do rats deprived of estrogen, but acquire egocentric or "response" learning tasks more slowly than do those deprived of hormone, suggesting that estrogen treatment may bias the strategy a rat is able to use to solve tasks. To determine if natural fluctuations in ovarian hormones influence cognitive strategy, we tested whether strategy use fluctuated across the estrous cycle in reproductively intact female rats. We found that in two tasks in which rats freely choose the strategy used to solve the task, rats were more likely to use place strategies at proestrous, that is, when ovarian steroids are high. Conversely, estrous rats were biased toward response strategies. The data suggest that natural fluctuations in ovarian steroids may bias the neural system used and thus the cognitive strategies chosen during learning and memory. PMID- 15109908 TI - Neurotherapeutic action of testosterone on hamster facial nerve regeneration: temporal window of effects. AB - Neurotherapeutic or neuroprotective effects of gonadal steroids on the injured nervous system have been demonstrated in our laboratory and others. We have previously demonstrated that testosterone propionate (TP) administered systemically at supraphysiological levels accelerates both recovery from facial paralysis and regeneration rates following facial nerve injury in the hamster. Initial temporal studies of steroidal enhancement of functional recovery from facial paralysis established that steroid exposure is necessary during the first postoperative week. Furthermore, accumulated evidence suggests that TP manifests its effects on neuronal regeneration in the immediate postoperative or preregenerative phase by altering the cellular stress response. The purpose of this study was to identify the effective temporal window of TP exposure sufficient to enhance regenerative properties of injured facial motoneurons and functional recovery from facial paralysis induced by facial nerve injury. Adult castrated male hamsters received a right facial nerve crush axotomy at the stylomastoid foramen and were divided into (1) short term, (2) delayed, (3) continuous, and (4) no TP treatment groups. Short term and continuous groups were implanted with 1 subcutaneous (sc) TP capsule each immediately after axotomy, with the capsule removed at 30 min, 2, 4, or 6 h in short-term groups and allowed to remain for the duration of the experiment in the continuous group. In the delayed TP group, 1 sc TP capsule was implanted 6 h after axotomy and allowed to remain for the duration of the experiment. For regeneration rate studies, postoperative times ranged from 4 to 7 days. For the behavioral studies, observations were made for 26 days postaxotomy. The results point to a critical 6 h interval immediately after injury when TP enhances nerve outgrowth distances and augments behavioral recovery. PMID- 15109909 TI - Multiple mechanisms of phenotype development in the bluehead wrasse. AB - Despite having detailed information on mechanisms mediating sex-typical behavior in many species, we have little understanding of whether the same mechanisms regulate these behaviors when they are performed in the same species under different social contexts. In the five field experiments of this study of bluehead wrasses (Thalassoma bifasciatum), a sex-changing fish, we examined the roles of arginine vasotocin (AVT) and the potent teleost androgen 11 ketotestosterone (11KT) in mediating sexual and aggressive behaviors typical of dominant males. We demonstrated that AVT appears necessary for the assumption of dominant territorial status in males and females, but is sufficient only in the socially dominant terminal phase (TP) male phenotype. Specifically, an AVP V(1) receptor antagonist prevented both TP males and females from gaining dominance over recently vacated territories. However, unlike TP males in a previous study, neither females nor initial phase males responded to AVT treatment with increases in display of TP male typical behaviors when under social conditions that inhibit sex change. Treating females with 11KT did not alter responsiveness to AVT, but did induce male coloration and courtship behavior that was not observed in oil treated females. Combined with the results of a previous study, these results indicate that the ability of AVT to induce male-typical behavior differs among sexual phenotypes and that this differential responsiveness appears to be dependent on social context and not directly on exposure to 11KT. Furthermore, since 11KT can induce courtship behavior in females that is not affected by AVT, there may be different hormonal mechanisms mediating courtship behavior under different social contexts. PMID- 15109910 TI - Both oxytocin and vasopressin may influence alloparental behavior in male prairie voles. AB - Neuropeptides, especially oxytocin (OT) and arginine vasopressin (AVP), have been implicated in several features of monogamy including alloparenting. The purpose of the present study was to examine the role of OT and AVP in alloparental behavior in reproductively naive male prairie voles. Males received intracerebroventricular (ICV) injections of artificial cerebrospinal fluid (aCSF), OT, an OT receptor antagonist (OTA), AVP, an AVP receptor antagonist (AVPA), or combinations of OTA and AVPA and were subsequently tested for parental behavior. Approximately 45 min after treatment, animals were tested for behavioral responses to stimulus pups. In a 10-min test, spontaneous alloparental behavior was high in control animals. OT and AVP did not significantly increase the number of males that showed parental behavior, although more subtle behavioral changes were observed. Combined treatment with AVPA and OTA (10 ng each) significantly reduced male parental behavior and increased attacks; following a lower dose (1 ng OTA/1 ng AVPA), males were less likely to display kyphosis and tended to be slower to approach pups than controls. Since treatment with only one antagonist did not interfere with the expression of alloparenting, these results suggest that access to either OT or AVP receptors may be sufficient for the expression of alloparenting. PMID- 15109911 TI - Expression of chemokine and receptors in Lewis rats with experimental autoimmune anterior uveitis. AB - The purpose of this study is to investigate the sequential expression of certain chemokines and chemokine receptors in the iris-ciliary body and popliteal lymph nodes of Lewis rats and, thus, to establish their roles in experimental autoimmune anterior uveitis. Uveitis was induced with the injection of melanin associated antigen intraperitoneally and into the left foot. The clinical severity of the uveitis was scored. At defined time points, CC chemokines (monocyte chemoattractant protein-1, macrophage inflammatory protein-1, and regulated-upon-activation normal T-cell expressed and secreted), CXC chemokines (interferon gamma-inducible protein-10, stromal-derived factor-1, and interleukin 8), and receptor (CCR2, CCR3, CCR5, CXCR1, CXCR2, CXCR3, and CXCR4) mRNA expression were semiquantified by using a reverse-transcriptase reaction followed by polymerase chain reaction. The concentrations of macrophage inflammatory protein-1 and regulated-upon-activation normal T-cell expressed and secreted in aqueous humor were determined by means of enzyme-linked immunosorbent assay. Levels of monocyte chemoattractant protein-1, macrophage inflammatory protein-1 and interferon gamma-inducible protein-10 started increasing before the clinical onset of disease; these might have been involved in the initial recruitment of inflammatory cells. The level of regulated-upon-activation normal T-cell mRNA, however, started rising concurrently with the onset of clinical disease, suggesting that this chemokine may exert amplifying role in generating uveitis. Stromal-derived factor-1 exhibited an early and high level of expression with the increase of cognate receptor, CXCR4, indicating that stromal-derived factor-1 plays a role in either promoting angiogenesis or attracting for T-cells. Instead of upregulation like other chemokine receptors, interleukin-8 receptors, CXCR1and CXCR2, mRNA could not be detected in accord with the increase of interleukin-8. These findings appeared that downregulation of chemokine receptors on neutrophils may make themselves less respond to interleukin-8 and subsequently lead to decreased recruitment of neutrophils into the iris-ciliary body. In addition, the expression of chemokine receptors in popliteal lymph nodes were earlier than those in the iris-ciliary body. This sequence of expression may reflect the process of T lymphocytes maturation and differentiation. Monocyte chemoattractant protein-1 protein was immunohistologically detected in the ciliary epithelium and infiltrating leukocytes. The above results suggest that chemokines, which act on T cells and monocytes, are sequentially upregulated during the clinical course of experimental autoimmune anterior uveitis, and thus, may contribute to the pathogenesis of acute anterior uveitis. PMID- 15109912 TI - Platelet derived growth factor (PDGF)-induced reactive oxygen species in the lens epithelial cells: the redox signaling. AB - Low level of reactive oxygen species (ROS) has been shown to play an important role in host defense and mediating mitogen-stimulated cell signaling in several cell types. This study is to identify the mitogen-induced endogenous ROS generation and the range of exogenous H(2)O(2) that initiate redox signaling and cell proliferation in human lens epithelial cells (HLE B3), using platelet derived growth factor (PDGF) as a model. To detect ROS generation, serum starved HLE cells (1.6 million) were loaded with fluorescent dye, 2',7' dichlorofluorescin diacetate (DCFH-DA), before exposing to PDGF (1 ng ml(-1)). The fluorescence generated from the oxidant-sensitive DCFH, the intracellular product of DCFH-DA hydrolysate, was immediately measured in live cells by confocal laser light microscopy (lambda(Ex)=488 nm, lambda(Em)=522 nm, laser power=10%). PDGF-stimulated cells showed strong transient fluorescence during the 60 min while no fluorescence could be seen in the unstimulated cells. The PDGF induced fluorescence could be suppressed with cells preloaded with N-acetyl-L cysteine (NAC, 30 mm), catalase (1 mg ml(-1)), or D-mannitol (100mm). The ability of catalase to penetrate and function in HLE cells was confirmed by western blot, enzyme activity and immunofluorescence microscopic analyses. PDGF induced DNA synthesis within one hour as measured by (3)H-thymidine incorporation, and transiently activated the mitogen-activated protein kinases (MAPKs) of ERK1/2 and JNK. PDGF-stimulated DNA synthesis and MAPK activation were eliminated in the presence of catalase or mannitol. Low levels of H(2)O(2) (10-20 microm) mimicked PDGF in both MAPK stimulation and cell proliferation. In conclusion, the mitogenic stimulus function of PDGF in HLE cells appears to be mediated via ROS to activate MAPKs and cell proliferation, which can be mimicked by low levels of H(2)O(2). It is proposed that the physiological function of ROS, the redox signaling, is present in the HLE cells and may play an important role in the development and maintenance of the lens. PMID- 15109913 TI - Melatonin protects human retinal pigment epithelial (RPE) cells against oxidative stress. AB - Oxidative stress is involved in the pathogenesis of age-related macular degeneration (AMD). Administration of conventional antioxidants has been shown to slow the progression of AMD and vision loss. Melatonin, an endogenous neurohormone produced by the pineal gland and retina, has been reported to be a potent antioxidant and free radical scavenger. In this study we tested whether melatonin can protect retinal pigment epithelial (RPE) cells against hydrogen peroxide (H(2)O(2))-induced cell death. Since mitochondrial DNA (mtDNA) is preferentially susceptible to oxidative damage, we tested whether melatonin can reduce H(2)O(2)-induced mtDNA lesions. A human RPE cell line (ARPE-19) was cultured and exposed to H(2)O(2) (100 and 200 microm) for 1 hr to induce cell death. Prior to H(2)O(2) treatment, cells were treated with various concentrations (0.1-200 microm) of melatonin for 2, 24 or 72 hr. Control cells received either melatonin or ethanol alone. Cell viability, as determined by MTT assay, showed no significant (P>0.05) protection against H(2)O(2) toxicity in cells receiving 2- and 24-hr pretreatment of melatonin at either concentration. However, when melatonin was administered diurnally for 3 consecutive days, this prolonged treatment markedly reduced H(2)O(2)-induced cell death (P>0.05) MtDNA damage, as assessed with quantitative PCR, was significantly decreased (P<0.05) in RPE cells pretreated with melatonin as compared to those without melatonin treatment. These results suggest that melatonin may play a role in protecting RPE cells from oxidative stress. PMID- 15109914 TI - Minocycline delays photoreceptor death in the rds mouse through a microglia independent mechanism. AB - PURPOSE: Minocycline, a semi-synthetic tetracycline antibiotic is reported to be neuroprotective in degenerative and ischaemic models of central nervous system disease, via mechanisms involving suppression of both cytotoxic microglial activity and caspase-dependent apoptosis. We have investigated the effect of minocycline treatment on a mouse model of retinitis pigmentosa, an inherited photoreceptor neurodegenerative disorder, and contrasted this with the effect of depleting retinal microglia using liposomal clodronate. METHODS: rds mice were treated intraperitoneally from the second postnatal day (P2) with either daily minocycline until P16, P18, P21, P24 and P27 or alternative day clodronate liposomes until P16. Immunohistochemical and immunofluorescent methods were applied for the detection of microglia (F4/80) and apoptosis (TUNEL and caspase 3 activation). RESULTS: Photoreceptor apoptosis was delayed by minocycline treatment but not, ultimately, prevented. Markedly reduced expression of activated caspase 3 was observed in photoreceptors at the early time point, corresponding with the reduced level of apoptosis. Delayed photoreceptor apoptosis due to minocycline treatment was associated with a 50% reduction in the numbers of microglia at early timepoints. Liposomal clodronate treatment also resulted in a marked reduction in the number of microglia (63% reduction in microglia), but in contrast to minocycline treatment, this had no effect on photoreceptor apoptosis. CONCLUSIONS: Minocycline appears to delay photoreceptor apoptosis through a microglia-independent action. Although microglial cytotoxicity has been implicated during other models of neurodegeneration, microglia are unlikely to play such a role in this model of photoreceptor dystrophy. PMID- 15109915 TI - Protein s-glutathionylation in retinal pigment epithelium converts heat shock protein 70 to an active chaperone. AB - A disulfide bond between key redox-sensitive cysteine residues and glutathione is one mechanism by which redox related allosteric effectors can regulate protein structure and function. Here we test the hypothesis that glutaredoxin-1 (Grx-1), a member of the oxidoreductase family of enzymes, may be a critical component of redox-sensitive molecular switches by mediating reversible protein S glutathionylation and enzymatic catalysis of thiol/disulfide exchange. Deglutathionylation of a 70 kDa protein by Grx-1 was detected using a monoclonal antibody specific to protein S-glutathionylation. Heat shock cognate protein 70 (Hsc70) was identified as a substrate of Grx-1 through mass spectrometry. Recombinant Hsc70 was glutathionylated in vitro, and protein S-glutathionylation reversed by Grx-1. Glutathionylated Hsc70 was more effective in preventing luciferase aggregation at 43 degrees C than reduced Hsc70 in a dose dependent fashion. ATP did not effect the chaperone activity of Hsc70-SG but did increase the activity of reduced Hsc70-SG. Reversible glutathionylation of Hsc70 may provide a mechanism for post-translation regulation of chaperone activity. PMID- 15109916 TI - Role of the microtubule cytoskeleton in traffic of EGF through the lacrimal acinar cell endomembrane network. AB - We have previously documented a novel biphasic traffic pattern for epidermal growth factor (EGF) in the acinar epithelial cell of the lacrimal gland. Different from the typical paradigm observed in many other cell types, EGF initially accumulates in the acinar basal-lateral recycling endosome, then is re directed to the prelysosomes and lysosomes and degraded. While the cellular content of intact EGF decreases by 40% between 20 and 120 m of continuous incubation at 37 degrees C, the EGF receptor (EGFR) content decreases only modestly [J. Cell Physiol. 199 (2004) 108]. The purpose of the present study was to investigate the role of the microtubule cytoskeleton in this traffic. Primary cultured rabbit lacrimocytes were incubated with [(125)I]-EGF, lysed, and analyzed by subcellular fractionation on sorbitol density gradients. Nocodazole treatment appeared to slightly decrease the initial uptake rate but to have no significant effect on the total amount of [(125)I] accumulation. However, it enhanced accumulation of [(125)I]-EGF and EGFR in the basal-lateral recycling endosome, and it enhanced accumulation of prepro- and pro- cathepsin B in fractions containing late endosomes and prelysosomes. Nocodazole permitted the time-dependent release of [(125)I]-EGF from the recycling endosome, but it partially inhibited [(125)I]-EGF degradation and decreased accumulation of [(125)I]-labeled degradation products in the lysosome. The microtubule-based molecular motors, cytoplasmic dynein and kinesin, were localized in compartments containing the late endosomes, prelysosomes, and lysosomes, consistent with the suggestion that microtubule-based molecular motors play important roles in traffic within the lysosomal pathway. Confocal fluorescence microscopy imaging of FITC-EGF substantiated the effects observed in biochemical studies by demonstrating that nocodazole increased accumulation in a peripheral compartment and decreased traffic to a perinuclear compartment. These data suggest that initial accumulation in the basal-lateral recycling endosome and subsequent release from the recycling endosome to the late endosomes and prelysosome are not microtubule-dependent. On the other hand, microtubule-based motors are more critical for traffic from the prelysosome to the lysosome. PMID- 15109917 TI - Recruitment of marrow-derived endothelial cells to experimental choroidal neovascularization by local expression of vascular endothelial growth factor. AB - PURPOSE: The question of whether adult animals maintain a reservoir of endothelial progenitor cells (EPCs) in the bone marrow that is involved in neovascularization is under investigation. The following study was undertaken to examine the potential contribution of EPCs to the development of choroidal neovascularization (CNV) in adult mice and to examine the role of local expression of vascular endothelial growth factor (VEGF) in this process. METHODS: Lethally irradiated, adult female nude mice were engrafted with whole bone marrow isolated from male transgenic mice expressing LacZ driven by the endothelial specific Tie-2 promoter. Two months, following bone marrow reconstitution, confirmed by quantitative Taqman PCR, an E1-deleted adenoviral vector expressing vascular endothelial growth factor (165) (Ad.VEGF(165)) was injected subretinally to induce CNV, confirmed by collagen IV immunohistochemistry. Bone marrow-derived endothelial cells were detected using either X-gal staining or Y chromosome in situ hybridization. Y chromosome positive cells within the CNV were confirmed to be endothelial cells by lectin staining. RESULTS: Subretinal Ad.VEGF(165) was capable of inducing CNV. Four-week old lesions were found to contain LacZ expressing cells within the CNV in bone marrow transplanted animals but not in negative control animals. Eighteen percent of all Y chromosome positive cells within the CNV were found to be lectin positive while 27% of all endothelial cells within the CNV were Y chromosome positive. CONCLUSION: Engrafted bone marrow-derived EPCs were shown to differentiate into endothelial cells at the site of subretinal VEGF-induced CNV in mice. These results suggest that EPCs contribute to the formation of neovascularization and that subretinal expression of VEGF might play an important role in recruitment of these cells to the site of CNV. PMID- 15109918 TI - Ultrahigh resolution optical coherence tomography of the monkey fovea. Identification of retinal sublayers by correlation with semithin histology sections. AB - Optical coherence tomography (OCT) has become an established diagnostic tool for the clinical assessment of retinal pathology but correlation of acquired signals with retinal substructures has often been ambiguous. In the monkey retina we have now obtained ultrahigh resolution (UHR) OCT images with 1.4 microm axial x 3 microm transverse resolution from perfusion-fixed eye cups of Macaca fascicularis and optimized the identification of retinal anatomy by correction of spatial artefacts in correlated histology. After resin embedding, serial semithin sections were obtained that corresponded to OCT transects. The direct overlay of features identified in histological sections with corresponding OCT locations was limited by non-linear tissue shrinkage due to dehydration and sectioning stress. In the present study, these misalignments were further corrected by using polygonal spline morphing based on corresponding unequivocal landmarks. The geometric normalization then allowed detailed comparison of both profiles including delicate sublayers of photoreceptor inner- and outer segments. Such correlation will facilitate the extraction of structural information from in vivo ultrahigh resolution OCT images in clinical and experimental applications. PMID- 15109919 TI - Hammerhead ribozyme targeting connective tissue growth factor mRNA blocks transforming growth factor-beta mediated cell proliferation. AB - PURPOSE: Excessive scarring following trauma or surgery of cornea, conjunctiva or retina can greatly impair visual outcome. At present, no agents are clinically available that selectively reduce activity of genes that regulate fibrosis. Connective tissue growth factor (CTGF) has been linked to fibrosis in several tissues, including cornea and conjunctiva. In this study, hammerhead ribozymes targeting CTGF mRNA were synthesized, kinetic parameters were measured, and the effect on TGF-beta-mediated cell proliferation was measured in cultured human fibroblasts. METHODS: The mRNA sequence of human CTGF was scanned for potential hammerhead ribozyme cleavage sites, and predicted secondary folding structures around the sites were calculated. Synthetic 12mer ribozymes and 33mer oligonucleotide mRNA targets corresponding to two sites were synthesized, and kinetic constants calculated from Hanes-Wolff plots of in vitro cleavage reactions. The ribozyme with higher percentage cleavage and kinetic rate was cloned into an expression plasmid (pTR-UF21) and stably transfected into cultured human fibroblasts. An inactive ribozyme plasmid served as a negative control. The effects of the ribozyme on expression of TGF-beta-induced CTGF mRNA and protein levels were measured using ELISA and real-time TaqMan quantitative RT-PCR. Finally, the effect of the CTGF ribozyme on TGF-beta-mediated proliferation of fibroblasts was measured using a non-radioactive cell proliferation microtiter assay. RESULTS: Of the eight potential hammerhead ribozyme cleavage sites in human CTGF mRNA, two sites (CHR 745, and CHR 859) were identified with optimal secondary folding. CHR 859 cleaved 94% of the target mRNA, compared to 46% cleavage for CHR 745 after 16 hr of reaction. CHR 859 had a K(m) of 1.56 microM and a K(cat) of 2.97 min(-1), while CHR 745 had a K(m) of 7.80 microM and a K(cat) of 5.7 min(-1). The turnover numbers (K(cat)/K(m)) of CHR 859 and CHR 745 were 1.9 x 10(6) M min(-1) and 7.4 x 10(5) M min(-1), respectively, indicating CHR 859 is 2.6 times more efficient than CHR 745 in destroying CTGF mRNA. Stable transfection of CHR 859 into human fibroblasts reduced CTGF mRNA levels 55% and protein levels 72% compared to the inactive ribozyme control. Furthermore, TGF beta-induced cell proliferation was reduced 90% in fibroblasts stably transfected with CHR 859 compared to control cell groups. CONCLUSIONS: The CHR 859 hammerhead ribozyme cleaved human CTGF mRNA with high kinetic efficiency in vitro, effectively reduced levels of CTGF mRNA and protein in cultured human fibroblasts, and blocked TGF-beta-induced cell proliferation without nonspecific toxicity. These data support the concept that CTGF mediates TGF-beta-induced cell proliferation, and imply that regulating CTGF expression with ribozymes may be effective in reducing ocular scarring. PMID- 15109920 TI - The presence of keratin 5 as an IgG Fc binding protein in human corneal epithelium. AB - Rabbit immunoglobulin G (IgG) binds specifically to the human corneal epithelium. We investigated this phenomenon to identify the binding material in the tissue. Sections of human cornea were immunostained with negative-control grade rabbit IgG, F(ab')(2) (the antigen recognition fragment of IgG yielded by pepsin digestion), and Fc fragments. Human corneal epithelium was homogenized in a buffer containing Triton X-100 and the water-insoluble residue was subjected to two-dimensional gel electrophoresis. Western blotting was performed on the gels to detect materials that bound with rabbit IgG Fc fragments. Spots in the gel that bound with the Fc fragments were collected and analysed by peptide mass fingerprinting (PMF). Using an antibody against a binding candidate, keratin 5 (suggested by PMF analysis), we checked the occurrence of this phenomenon. Immunohistochemistry showed that rabbit IgG bound to corneal epithelium via the Fc region but not F(ab')(2). Western blots using rabbit Fc fragments showed that some protein spots in the gel bound to the fragments. All of the binding materials were basic and had molecular weights of approximately 55 kDa. PMF analysis showed that the binding material was keratin 5; all binding materials on the transferred membrane were recognized by an anti-keratin 5 antibody. On corneal sections, anti-keratin 5 produced staining similar to that of rabbit IgG. These findings suggest that rabbit IgG binds to keratin 5 in the human corneal epithelium via the Fc region of the molecule. PMID- 15109921 TI - The role of opsin expression and apoptosis in determination of cone types in human retina. AB - In primates, short wavelength sensitive cones (S cones) and medium- or long wavelength-sensitive cones (L/M cones) are two separate populations. Each cone type has a different developmental timecourse, contributes to different intra retinal circuits, and transmits different types of information to the brain. However, in fetal human retina a significant population of cones express both S and L/M opsin (S+L/M cones), raising questions about whether S+L/M cones die or change opsin expression during development. We have utilized fetal, postnatal and adult human retinae to study the immunohistochemical distribution and morphology of S+L/M cones during development. Because S cones appear to be at higher density in fetal compared to adult retinae, we used antibodies to S opsin and alpha transducin to estimate the proportion of S-cones, and TUNEL labelling to detect apoptotic death in the L/M, S or S+L/M population during development. S cones were present in central retina from fetal week (Fwk)11 and covered the retina by Fwk20. L/M cones appeared in the foveal cone mosaic 3-4 weeks after S-opsin was first detected, and covered the retina by birth. S+L/M cones were detected in all retinae older than Fwk14. They were most numerous at the retinal eccentricity where L/M opsin was just appearing; i.e. at the 'front' of L/M opsin expression. In this region, five morphological types of cones were present. (1) Heavily labelled S cones had thick cell bodies, a thick basal axon and pedicle, and a nucleus at any level of the outer nuclear layer (ONL). (2) Heavily labelled L/M cones were wine goblet shaped with a small round cell body, a large nucleus at the outer ONL edge, and a thin axon with a prominent synaptic pedicle. (3) Goblet shaped S+L/M cones. (4) Goblet-shaped cones lightly labelled for S-opsin. (5) Cones that were not immunoreactive to either opsin. Only type 1 S cones were present peripheral to the L/M expression front, and their labelling intensity, morphology and distribution indicates that these are the 'true blue' cones of the adult mosaic. Only type 2 L/M cones were present in the foveal cone mosaic. Types 3 and 4 were most numerous within 500-750 microm of the L/M expression front, but type 3 S+L/M cones were also scattered throughout more central regions in fetal, infant and adult retinae. S+L/M cones comprised 5-10% of opsin immunoreactive cones at the L/M front in fetal and early postnatal retinas but 0.01-0.03% throughout P8mo and adult retinae. We found no evidence of significant levels of apoptosis in L/M cones at the expression front, suggesting that this decrease was not due to cell death. The findings suggest that goblet-shaped cones destined to express L or M opsin may initially and transiently express S opsin. Near the optic disc, at Fwk17 S cone density was around 2000 cells mm(-2), which dropped 50% by Fwk20 and stabilized at around 500 cells mm(-2) by birth. Double labelling with alpha-transducin showed that throughout this period 8-10% of all cones expressed S opsin. TUNEL labelling found no significant apoptosis in the S cone population. The decrease in S cone density near the optic disc occurs in the absence of apoptosis, and is likely due to other developmental events acting on the photoreceptor layer, including displacement of cones towards the fovea. PMID- 15109922 TI - TIMP-1 role in protection against Pseudomonas aeruginosa-induced corneal destruction. AB - To establish the role of TIMP-1 in protection against Pseudomonas aeruginosa induced corneal destruction, corneas of adult 8-week-old (resistant) and aged 12 month-old (susceptible) mice were infected with the bacterium. Corneas were analyzed for TIMP-1 protein by immunocytochemistry and Western blotting. Basement membrane (BM) integrity was assessed by immunostaining for type IV collagen. Additionally, resistant 8-week-old mice were treated systemically with neutralizing TIMP-1 polyclonal antibody (pAb) or pre-immune normal rabbit serum (NRS). Ocular and BM integrity as well as MMP-9 expression were examined in these mice. A greater amount of TIMP-1 protein was observed in the cornea of 8-week-old mice. In the cornea, the strongest staining was found in the superficial epithelium, but positive staining also was seen in the basal epithelium and stroma. When type IV collagen was analyzed in the BM of both age groups of mice, a distinct staining pattern was observed in only the young adult mice. Treatment of 8-week-old resistant mice with neutralizing TIMP-1 pAb vs NRS increased the amount of MMP-9 in the cornea of TIMP-1 pAb-treated mice and affected the ability of these mice to deposit BM components. These studies suggest that adequate expression of TIMP-1 protects against BM and stromal degradation via multiple processes. PMID- 15109923 TI - Persistent respiratory changes following intermittent hypoxic stimulation in cats and human beings. AB - Repeated intermittent hypoxia or other stimulation of carotid chemoreceptors produces a consistent long-term increase in respiratory nerve activity in vagotomized, artificially ventilated anesthetized or decerebrate animals, but variable results have been reported in more intact preparations. We sought additional variables that could be measured to help gain an understanding of persistent respiratory responses to intermittent hypoxia. The variance of respiratory phases decreased in 10 of 11 recordings from vagotomized anesthetized cats during long-term facilitation induced by carotid chemoreceptor stimulation. The variance of expiratory time was reduced in 10 awake human beings exposed to repetitive, brief episodes of isocapnic hypoxia (6% O(2) in N(2), 60s). Respiratory frequency was increased in humans and tidal volume decreased so that minute ventilation remained unchanged. The results suggest that there are persistent changes in the output of the respiratory central pattern generator following intermittent peripheral chemoreceptor stimulation or hypoxia. PMID- 15109924 TI - Effects of inactivation and stimulation of locus coeruleus on respiratory activity of neonatal rat. AB - We examined the effects of pharmacological inactivation and electrical stimulation of the locus coeruleus (LC) on the respiratory rate (RR) in isolated brainstem-spinal cord preparations of the neonatal rat. The brainstem and spinal cord were isolated en bloc from neonatal (days 1-4) Sprague-Dawley rats and superfused with artificial cerebrospinal fluid (ACSF) equilibrated with a gas mixture containing 2% CO(2) (pH 7.8). Pharmacological inactivation of the bilateral LC by means of microinjection of tetrodotoxin or noradrenaline elicited a significant decrease in RR in preparations obtained from rats aged 3-4 days, but not in preparations of rats aged 1-2 days. Stimulation of the bilateral LC with a train of electrical pulses (25-50 microA, 30 ms, 0.5 Hz, 3-4 min) caused a marginal but significant increase in RR on days 3-4, but not on days 1-2. These results indicate that the LC exerts an excitatory effect on the medullary respiratory rhythm generator in an age-dependent manner. PMID- 15109925 TI - Hypoxic response in newborn rat is attenuated by neurokinin-1 receptor blockade. AB - Substance P (SP) is considered to be involved in the regulation of respiration, in particular when respiratory demands are increased, such as during hypoxic stress. In the present study we have investigated the effects of intracerebroventricular pre-treatment with the selective NK-1 receptor antagonist RP67580 on the respiratory response to hypoxia in 5-day-old rat pups. Basal respiration was not altered by RP67580. When subjected to hypoxia (10% O(2)), rat pups pre-treated with RP67580 were unable to sustain the increased respiratory frequency at 10 min. In situ hybridisation demonstrated increased expression of c fos mRNA in several brainstem areas following hypoxia. This activation was blocked by the antagonist in the retrotrapezoid nucleus and the rostral ventrolateral medulla, areas known to be involved in the hypoxic ventilatory response. This study corroborates a role of endogenously released SP, mediated via NK-1 receptors, in the sustained response to hypoxia in 5-day-old rat pups and suggests that neurons in the rostral ventrolateral medulla are important in this function. It also represents a further example that neuropeptides are released under stressful conditions. PMID- 15109926 TI - Day/night pattern of arterial blood gases in the cow. AB - Both metabolic rate and pulmonary ventilation change throughout 24h with a circadian pattern. Because their changes occur almost in synchrony and by a similar amount, blood gases may remain steady within a narrow range. We tested this possibility in five cows (Bos taurus, Bruna Italiana breed), maintained in a stable at 29 degrees C, under natural light-dark (LD) regime, by measuring arterial blood gases every 3h for 2 days. All cows presented a clear day/night pattern of body temperature (T(b)), with an average peak-trough difference (PTD) of 0.5 degrees C. Breathing rate oscillated significantly in three out of five animals, with a group-mean PTD of 2 breaths per minute, and it was time-advanced with respect to the oscillation of T(b). Significant oscillations in arterial pH, bicarbonate, partial pressure of oxygen and partial pressure of carbon dioxide (Pa(CO2) ) were observed in, respectively, 1 cow out of 5, 1/5, 3/5 and 5/5. Of all these variables, group-mean analysis revealed a significant day/night pattern only for Pa(CO2), and even in this case the average PTD was less than 1 mmHg. We conclude that, in the cow, blood gases remain remarkably stable throughout the 24 h. Hence, the daily oscillations of body temperature, breathing rate, and probably of many other factors affecting metabolic rate and pulmonary ventilation do not preclude an excellent homeostasis of blood gases. PMID- 15109927 TI - Low-dose acetazolamide reduces the hypoxic ventilatory response in the anesthetized cat. AB - Low intravenous dose acetazolamide causes a decrease in steady-state CO(2) sensitivity of both the peripheral and central chemoreflex loops. The effect, however, on the steady-state hypoxic response is unknown. In the present study, we measured the effect of 4 mg x kg(-1) acetazolamide (i.v.) on the isocapnic steady-state hypoxic response in anesthetized cats. Before and after acetazolamide administration, the eucapnic steady-state hypoxic response in these animals was measured by varying inspiratory P(O2) levels to achieve steady-state Pa(O2) levels between hyperoxia Pa(O2) approximately 55 kPa, approximately 412 mmHg) and hypoxia (Pa(O2) approximately 7 kPa, approximately 53 mmHg). The hypoxic ventilatory response was described by the exponential function V(I) = G exp (-DP(o2) + A with an overall hypoxic sensitivity G, a shape parameter D and ventilation during hyperoxia A. Acetazolamide significantly reduced G from 3.057 +/- 1.616 to 1.573 +/- 0.8361 min(-1) (mean +/- S D). Parameter A increased from 0.903 +/- 0.257 to 1.193 +/- 0.321 min(-1), while D remained unchanged. The decrease in overall hypoxic sensitivity by acetazolamide is probably mediated by an inhibitory effect on the carotid bodies and may have clinical significance in the treatment of sleep apneas, particularly those cases that are associated with an increased ventilatory sensitivity to oxygen and/or carbon dioxide. PMID- 15109928 TI - Hypoxia of sleep apnoea: cardiopulmonary and cerebral changes after intermittent hypoxia in rats. AB - Sleep apnoea (SA) is common, especially in elderly people. In severe cases, arterial P(O2) may be lowered for a third or more in a night of sleep. To simulate the degree and duration of severe SA we exposed rats in a normobaric environmental chamber to 10% O(2) for 4h daily for 56 days (intermittent hypoxia: IH group) and compared them with rats continuously exposed for 8 weeks (continuous hypoxia: CH group) and control rats breathing room air (normoxic: N group). We found significant cardiopulmonary and cerebral changes. Right ventricular hypertrophy developed in IH and to a greater extent in CH. Small peripheral lung vessels developed thicker walls (assessed by a new method), which reduced their lumen, more in CH than IH. Coronal brain sections were immunostained for the glucose-transporter 1 (GLUT1) and the vascular endothelial growth factor (VEGF). The percentages of immunoreactivity in the frontal and temporal cortex, hippocampus, accumbens and putamen were determined by image capture analysis. We noted GLUT1 immunoreactivity of the capillaries was similarly increased in all regions after CH but less so after IH. However, there was a significant linear trend in GLUT1 reactivity from N to IH to CH (R(2) = 0.73, P = 0.007) that was also confirmed by analysis of variance. The extent of VEGF-stained neurones and glial cells was significantly increased in all regions after IH but not after CH. This suggests that the signals for angiogenesis were complete or arrested after CH. Our findings have implications for the elderly subjected to hypoxic episodes during sleep apnoea. PMID- 15109929 TI - Interactive effects of mechano- and chemo-receptor inputs on cardiorespiratory outputs in the toad. AB - Arterial blood pressure (P(b)), pulmocutaneous blood flow (Q(pc)), heart rate (f(H)), and fictive ventilation (motor activity in the Vth cranial nerve, V(int)), were recorded from decerebrated, paralysed toads receiving unidirectional ventilation with experimental gas mixtures over a range of lung inflation. At the onset of spontaneous bouts of fictive ventilation, (Q(pc)) and P(b) increased immediately, often with changes in heart rate, implying central cardiorespiratory interactions. Inflation of the lungs with different gas mixtures revealed that the effect of hypercarbia on V(int) was reduced by lung inflation and that feedback from pulmonary stretch receptors may summate with central feedforward control of f(H) and (Q(pc)) in an interactive fashion. The results of bolus injections of cyanide into the carotid or the pulmonary circulations suggest there are oxygen sensitive receptors in both circuits that affect the cardiovascular system directly and respiratory activity by complex central interactions with inputs from central chemoreceptors and pulmonary stretch receptors. PMID- 15109930 TI - Arousal and ventilatory responses to hypoxia in sleeping infants: effects of maternal smoking. AB - Our aim was to determine whether maternal cigarette smoking affects arousal and ventilatory responses to hypoxia in infants. Infants born to non-smoking (NS, n = 15) and smoking mothers (SM, n= 9) were studied at 2-5 weeks, 2-3 and 5-6 months. Ventilatory responses to 15% O(2) were determined preceding arousal. At each age and in both groups, infants aroused more frequently and earlier to hypoxia in active sleep (AS) than quiet sleep (QS). Arousal latency was longer in SM infants (in QS) at 5-6 months (P < 0.05). Baseline respiratory parameters were not different between groups, except that, at 2-3 months, SM infants had higher SP(O2) during AS than NS infants. Maternal smoking did not affect ventilatory responses preceding hypoxia-induced arousal in either sleep-state at any age. We conclude that mild hypoxia stimulates ventilation and arousal in infants up to 6 months and that arousability is depressed in SM infants at 5-6 months; however, ventilatory responses preceding arousal are not adversely affected by smoking. PMID- 15109931 TI - Yersinia YopJ inhibits pro-inflammatory molecule expression in human bronchial epithelial cells. AB - Human bronchial epithelial cell pro-inflammatory molecule expression plays a role in the pathogenesis of airway diseases. We hypothesize that Yersinia outer protein-J (YopJ), a Yersinia virulence effector which inhibits mitogen activated protein (MAP) kinase kinases (MKKs), attenuates epithelial cell pro-inflammatory molecule expression. 16HBE14o-cells were co-transfected with cDNAs encoding Yersinia pseudotuberculosis YopJ or empty vector. Expression of YopJ reduced activation of extracellular signal regulated kinase (ERK)-2, Jun amino terminal kinase (JNK)-1 and IkappaB kinase (IKK)-beta. YopJ also blocked transactivation of NF-kappaB and AP-1 promoter sequences which has been shown to regulate chemokine expression. Finally, expression of YopJ reduced transcription from the IL-8, RANTES (regulated upon activation, normal T cell expressed and secreted) and intercellular adhesion molecule (ICAM)-1 promoters. We conclude that YopJ expression blocks the innate immune response in lung epithelial cells, the site of Yersinia pestis infection. Inhibition of bronchial epithelial cell responses by YopJ is consistent with the notion that MAP kinases regulates bronchial epithelial cell pro-inflammatory molecule expression. PMID- 15109932 TI - Revised one-step method for determination of cardiac output. AB - Cardiac output (Q) is a determinant of blood pressure and O(2) delivery and is critical in the maintenance of homeostasis, particularly during environmental stress and exercise. Cardiac output can be determined invasively in patients; however, indirect methods are required for other situations. Soluble gas techniques are widely used to determine (Q). Historically, measurements during a breathhold, prolonged expiration and rebreathing to CO(2) equilibrium have been used; however, with limitations, especially during stress. Farhi and co-workers developed a single-step CO(2) rebreathing method, which was subsequently revised by his group, and has been shown to be reliable and compared closely to direct, invasive measures. V(CO2), P(ACO2), and P(VCO2) are determined during a 12-25s rebreathing, using the appropriate tidal volume, and (Q) is calculated. This method can provide accurate data in laboratory and field experiments during exercise, increased or decreased gravity, water immersion, lower body pressure, head-down tilt, altered ambient pressure or changes in inspired gas composition. PMID- 15109933 TI - Evidence that venoconstriction reverses the phase II sympathoinhibitory and bradycardic response to haemorrhage. AB - Severe hypotensive haemorrhage results in a biphasic response, characterized by an initial increase in heart rate and sympathetic vasomotor activity (phase I) followed by a life-threatening hypotension, accompanied by profound sympathoinhibition and bradycardia (phase II). The phase II response is believed to be dependent on inputs from cardiopulmonary receptors, and may be triggered by the reduction in venous return and cardiac filling associated with severe haemorrhage. In this study, we tested the hypothesis that the phase II response could be reversed by venoconstriction, which is known to enhance venous return and cardiac filling, by comparing the effects of phenylephrine (which constricts veins as well as arterioles) with that of vasopressin (which constricts arterioles but not veins). In sodium pentobarbitone-anaesthetised rats, haemorrhage evoked an initial increase in heart rate (HR) and renal sympathetic activity (RSNA) followed by a large decrease in both variables to levels below the pre-haemorrhage baseline levels (phase II response). During the phase II response, an intravenous injection of phenylephrine, sufficient to restore mean arterial pressure to the pre-haemorrhage level, resulted in a gradually developing increase (over 3-4 min) in HR and RSNA back to the baseline levels. In contrast, intravenous injection of an equipressor dose of vasopressin did not result in any increase in RSNA and only a transient increase in HR. Injection of phenylephrine, but not vasopressin, also increased the pulsatile component of central venous pressure, indicative of reduced venous capacitance. The findings indicate that venoconstriction reverses the phase II sympathoinhibition and bradycardia. PMID- 15109934 TI - GABA-mediated attenuation of noradrenaline release in the rat median preoptic area caused by intravenous injection of metaraminol. AB - Previous studies have shown that the noradrenergic system in the median preoptic nucleus (MnPO) play an important role in the control of the body fluid balance and cardiovascular function and that the release of noradrenaline in the MnPO is regulated by gamma-aminobutyric acid (GABA) receptor mechanisms. The present study was carried out to examine whether the GABAergic system is involved in the modulation of the noradrenaline release in the MnPO in response to an elevation in blood pressure using in vivo microdialysis techniques. In urethane anaesthetised male rats, the rise in arterial pressure caused by intravenous administration of the alpha-agonist metaraminol significantly decreased dialysate noradrenaline concentration in the MnPO area. The decrease in the noradrenaline level elicited by the metaraminol administration was significantly attenuated by perfusion with either bicuculline (10 microM), a GABA(A) receptor antagonist, or phaclofen (10 microM), a GABA(B) receptor antagonist, through a microdialysis probe. The amount of the antagonist-induced attenuation was much greater in the bicuculline-treated group than in the phaclofen-treated group. These results suggest that the release of noradrenaline in the MnPO area may be modulated by neural inputs from the peripheral baroreceptors, and that the neural inputs may be mediated in part through GABA(A) receptors rather than GABA(B) receptors in the MnPO area. PMID- 15109935 TI - Multiple signal pathways coupling VIP and PACAP receptors to calcium channels in hamster submandibular ganglion neurons. AB - The Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase activating polypeptide (PACAP) are two novel neuropeptides which produce particular biological effects caused by interaction with G-protein-coupled receptors. We have shown in a previous study where VIP and PACAP 38 inhibit voltage-dependent calcium channel (VDCC) currents (ICa) via G-proteins in hamster submandibular ganglion (SMG) neurons. In this study, we attempt to further characterize the signal transduction pathways of VIP-and PACAP 38-induced modulation of ICa. Application of 1 microM VIP and PACAP 38 inhibited ICa by 33.0 +/- 3.1% and 36.8 +/- 2.6%, respectively (mean +/- S.E.M., n = 8). Application of strong voltage prepulse attenuated PACAP 38-induced inhibition of ICa. Pretreatment of cAMP dependent protein kinase (PKA) activator attenuated VIP induced inhibition, but not the PACAP 38-induced inhibition. Intracellular dialysis of the PKA inhibitor attenuated the VIP-induced inhibition, but not the PACAP 38-induced inhibition. Pretreatment of protein kinase C (PKC) activator and inhibitor attenuated VIP-induced inhibition, but not the PACAP 38-induced inhibition. Pretreatment of cholera toxin (CTX) attenuated PACAP 38-induced inhibition of ICa. These findings indicate that there are multiple signaling pathways in VIP and PACAP 38-induced inhibitions of ICa: one pathway would be the VPAC1/VPAC2 receptors-induced inhibition involving both the PKA and PKC, and another one concerns the PAC1 receptor-induced inhibition via Gs-protein betagamma subunits. The VIP-and PACAP 38-induced facilitation of ICa can be observed in the SMG neurons in addition to inhibiting of ICa. PMID- 15109936 TI - Morphology of parasympathetic neurons innervating rat lingual salivary glands. AB - Saliva is essential for taste function and not only does saliva influence taste reception, but also taste perception initiates salivation. As a first step in investigating circuits involved in gustatory-salivary reflexes, we have studied the morphology of the rat inferior salivatory nucleus (ISN), which contains parasympathetic secretomotor neurons that control the parotid and lingual (von Ebner) salivary glands. By applying the fluorescent label Fluorogold to the cut end of the glossopharyngeal nerve, the neurons supplying only the lingual salivary glands were labeled. Confocal microscopy and three-dimensional reconstruction were used to analyze the labeled neurons in the horizontal plane to determine their morphological characteristics. Additional neurons were studied in the coronal plane to determine the influence of the plane of section on neuron morphology. Reconstructions indicated that inferior salivatory neurons extend in a rostral-caudal distribution just adjacent to the medial border of the nucleus of the solitary tract (NST). There is considerable morphological variability among neurons, with neurons having up to 6 primary dendrites and 17 dendritic segments that extend a maximum of 834 microm from the soma. However, although ISN neurons vary in the size and complexity of their dendritic trees, distributions of all measures of neuron morphology are unimodal, indicating that distinct groups of neurons are not revealed based on these measures. There is, however, variability in the orientation pattern of the dendritic trees that is not represented in either the population or mean measures. Individual neurons can be categorized with either mediolateral, rostro-caudal or no apparent preferred orientation. Comparisons of neurons in rostral, intermediate or caudal third of the ISN revealed regional differences in neuron morphology; neurons in the caudal third have significantly longer dendrites than those in the intermediate or rostral third. Thus, while ISN neurons belong to a single morphological grouping, they vary in the size and complexity of their dendritic trees, as well as having different dendritic orientations within the salivary nucleus. PMID- 15109937 TI - Spinal cord activation differentially modulates ischaemic electrical responses to different stressors in canine ventricles. AB - Spinal cord stimulation (SCS) represents an acceptable treatment modality for patients with chronic angina pectoris refractory to standard therapy, but its mechanism of action remains unclear. To develop an experimental paradigm to study this issue, ameroid (AM) constrictors were implanted around the left circumflex coronary artery (LCx) in canines. Six weeks later, unipolar electrograms were recorded from 191 sites in the LCx territory in the open-chest, anesthetized state under basal pacing at 150 beats/min. We investigated the effect of SCS on ST segment displacements induced in the collateral-dependent myocardium in response to two stressors: (i) transient bouts of rapid ventricular pacing (TRP: 240/min for 1 min) and (ii) angiotensin II administered to right atrial neurons via their coronary artery blood supply. ST segment responses to TRP consisted of ST segment elevation in central areas of the LCx territory and ST depression at more peripheral areas. Such responses were unchanged when TRP was applied under SCS. Shortening of repolarization intervals in the metabolically compromised myocardium in response to TRP was also unaffected by SCS. In contrast, ST segment responses to intracoronary angiotensin II, which consisted of increased ST elevation, were attenuated by SCS in 6/8 preparations. The modulator effects of SCS were greatest at sites at which the greatest responses to angiotensin II occurred in the absence of SCS. These data indicate that spinal cord stimulation may attenuate the deleterious effects that stressors exert on the myocardium with reduced coronary reserve, particularly stressors associated with chemical activation of the intrinsic cardiac nervous system. PMID- 15109938 TI - The defecation reflex in rats: fundamental properties and the reflex center. AB - While pharmacological and physiological studies in rats are now increasing, physiological properties of their defecation have been scarcely investigated. This study was performed to define the properties of defecation in decerebrate rats, with special reference to the pontine defecation reflex center, which has been postulated in dogs. Intraluminal pressure was recorded from the colon and rectum with balloon-pressure transducer method using balloons of 15-20 mm in length and 0.1-0.3 ml in volume. Distention of a balloon in the descending colon and rectum with an additional injection of 0.03-0.1 ml air induced propulsive contractions on the descending colon and rectum. The mean of threshold pressures to induce propulsive contraction was 17.0 +/- 5.8 mm Hg (mean+/-S.E.) in the proximal part and 18.3 +/- 3.3 mm Hg in the distal part of the descending colon, and 11.8 +/- 1.3 mm Hg in the rectum. The maximum amplitude of propulsive contractions was 55 mm Hg in the rectum, 47 mm Hg in the distal part of the descending colon and 38 mm Hg in the proximal part. Similar colorectal propulsive contractions were produced by gastric distention (5-10 ml, 20-30 mm Hg) and electrical stimulation of the anal canal. Contrarily, spontaneous contractions of the proximal colon were suppressed by rectal distention and anal-canal stimulation. These results suggest that the descending colon and rectum, but not the proximal colon, were innervated by the pelvic afferent and efferent fibers mediating the defecation reflex. Pontine transection at the cerebellar peduncle level abolished colorectal propulsive contractions induced by distention of the stomach, descending colon and rectum, and stimulation of the anal canal, although much smaller contractions were still induced after the pontine transection. These results suggest that the pontine defecation reflex center exists and works in rats, as in dogs. PMID- 15109939 TI - Prevention and partial reversal of diabetes-induced changes in enteric nerves of the rat ileum by combined treatment with alpha-lipoic acid and evening primrose oil. AB - Treatment with alpha-lipoic acid (LA) or evening primrose oil (EPO), individually, fails to prevent diabetes-induced changes in enteric nerves. Since synergy between these treatments has been reported, the aim was to investigate the effectiveness of combined LA/EPO treatment. LA and EPO were administered in the diet (approximately 80 and 200 mg/kg/day, respectively) to control and diabetic (induced by streptozotocin, 65 mg/kg, i.p.) rats. For prevention, treatment started after 1 week and lasted 7 weeks. For reversal, treatment lasted 4 weeks and was initiated after 8 weeks. Nerves supplying the ileum containing vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP) and noradrenaline (NA) were examined immunohistochemically or biochemically. Diabetes caused a significant increase in VIP-containing cell bodies (p<0.001), decrease in NA content (p<0.01) and loss of CGRP-immunoreactivity. LA/EPO treatment totally prevented diabetes-induced changes in VIP (p<0.001) and CGRP and partially reversed (p<0.05) these changes once they had been allowed to develop. In contrast, treatment had no effect on diabetes-induced changes in NA containing nerves. Therefore, LA and EPO are only effective at treating diabetes induced changes in some enteric nerves when administered in combination. However, diabetes-induced changes in NA-containing nerves are resistant to treatment. PMID- 15109940 TI - Influence of arginine vasopressin receptors and angiotensin receptor subtypes on the water intake and arterial blood pressure induced by vasopressin injected into the lateral septal area of the rat. AB - In this study we investigated the influence of d(CH2)5-Tyr(Me)-[Arg8]vasopressin (AAVP) and [adamanteanacetyl1,0-ET-d-Tyr2,Val4,aminobutyryl6,Arg8,9] [Arg8]vasopressin (ATAVP), which are antagonists of vasopressin V1 and V2 receptors, and the effects of losartan, a selective angiotensin AT1 receptor antagonist, and CGP42112A, a selective AT2 receptor antagonist, injected into the lateral septal area (LSA) on thirst and hypertension induced by [Arg8]vasopressin (AVP). AAVP and ATAVP injected into the LSA reduced the drinking responses elicited by injecting AVP into the LSA. Both the AT1 and AT2 ligands administered into the LSA elicited a concentration-dependent decrease in the water intake induced by AVP injected into the LSA, but losartan was more effective than CGP42112A. The increase in MAP, due to injection of AVP into the LSA, was reduced by prior injection of AAVP from 18 +/- 1 to 6 +/- 1 mm Hg. Losartan injected into the LSA prior to AVP reduced the increase in MAP to 7 +/- 0.8 mm Hg. ATAVP and CGP42112A produced no changes in the pressor effect of AVP. These results suggest that the dipsogenic effects induced by injecting AVP into the LSA were mediated primarily by AT1 receptors. However, doses of losartan were more effective when combined with CGP42112A than when given alone, suggesting that the thirst induced by AVP injections into LSA may involve activation of multiple AVP and angiotensin II receptor subtypes. The pressor response of AVP was reduced by losartan and by AAVP. CGP42112A and ATAVP did not change the AVP pressor response. These results suggest that facilitator effects of AVP on water intake are mediated through the activation of V1 receptors and that the inhibitory effect requires V2 receptors. The involvement of AT1 and AT2 receptors can be postulated. Based on the present findings, we suggest that the AVP in the LSA may play a role in the control of water and arterial blood pressure balance. PMID- 15109941 TI - Scavengers beware! PMID- 15109942 TI - Suppressing the suppressors. PMID- 15109943 TI - Germ stem cells produce oocytes in adult female mice. PMID- 15109944 TI - Rumour has it--a tale of idioms. PMID- 15109945 TI - Ligand efficiency: a useful metric for lead selection. PMID- 15109946 TI - Promiscuity: what protects us, perplexes us. PMID- 15109947 TI - Obituary of Dr Paul Janssen (1926-2003). PMID- 15109948 TI - Patch clamping by numbers. PMID- 15109949 TI - Orphan nuclear receptor-mediated xenobiotic regulation in drug metabolism. PMID- 15109950 TI - Unconventional natural sources for future drug discovery. PMID- 15109955 TI - Thrombin activates envelope glycoproteins of HIV type 1 and enhances fusion. AB - To elucidate the roles of serine proteases, including thrombin, in HIV infection, we treated H9 cells infected with HIV-1 LAI virus (H9/IIIB) with four different proteases (thrombin, cathepsin G, trypsin and chymotrypsin) and observed their effects on functional epitopes on both gp120 and gp41 by using flow cytometry. Monoclonal antibodies (MAbs) against the V3 loop, V2 loop, CD4 binding site, coreceptor binding site and gp41 were used. It was found that trypsin decreased the binding of all MAbs except for one MAb against the V3 loop (IIIB-V3-21). Chymotrypsin and cathepsin G did not show any remarkable effect on the antigen expression. On the other hand, thrombin decreased the reactivities of two out of four anti-V3 MAbs and increased the exposure of functional gp120 epitopes including the coreceptor binding site and CD4 binding site. Thrombin also increased the expression of 2F5 antigen (a neutralizing epitope of gp41) but had no effect on other gp41 epitopes. The effect of trypsin or thrombin on HIV induced cell fusion was examined through co-culturing H9/IIIB and MAGI cells. Trypsin slightly inhibited fusion. Fusion was significantly enhanced in a dose dependent manner by thrombin, and a 280% increase at 5 U/ml (P < 0.001) was observed. In conclusion, thrombin, one of the major inflammatory molecules in blood, facilitates HIV-induced cell fusion, probably by activating gp120. PMID- 15109956 TI - Specific reactions between purified HIV-1 particles and CD4+ cell membrane fragments in a cell-free system of virus fusion or entry. AB - The initial step of human immunodeficiency virus type 1 (HIV-1) infection has been studied by Env-mediated fusion or entry assays with appropriate cells expressing CD4 or CXCR4/CCR5 receptors in cultures, where many factors underlying cellular activities likely regulate the fusion/entry efficiency. Here we attempted to develop a more simplified in vitro cell-free fusion/entry reaction that mimics HIV-1 infection in cultures. Membrane fragments of target cells and intact infectious HIV-1 particles were purified, mixed and incubated. The core p24 protein was released from the purified virions and detected by ELISA without detergents in the supernatant of the reaction mixtures. This release reaction proceeded temperature-dependently and in a dose-dependent manner between the virion and membrane fractions, and was specific for HIV-1 Env and CD4. Env deleted or VSV-G-pseudotyped HIV-1 released little p24, if any. Pretreatment of the membrane fragments with anti-CD4 antibodies inhibited the p24 induction from both X4-tropic and R5-tropic HIV-1. Furthermore, X4 but not R5 HIV-1 reacted with the membrane prepared from intrinsically CXCR4-positive HeLa-CD4 cells, whereas both viruses reacted with that prepared from CCR5-transduced HeLa-CD4 cells, indicating that this cell-free reaction mimics coreceptor usage of HIV-1 infection. Therefore, a potent entry inhibitor of X4 HIV-1, SDF-1alpha, blocked the release from X4 but not R5 HIV-1. Inversely, a weak entry inhibitor of R5 HIV 1, MIP-1beta, partially affected only the release from R5 HIV-1. These results suggest that this cell-free reaction system provides a useful tool to study biochemical fusion/entry mechanisms of HIV-1 and its inhibitors. PMID- 15109957 TI - Characterization of Epstein-Barr virus infection in a human signet ring cell gastric carcinoma cell line, HSC-39. AB - In order to study the mechanism of Epstein-Barr virus (EBV) infection in gastric carcinoma cells, we characterized the EBV infection in signet ring cell line HSC 39, derived from a human gastric carcinoma. HSC-39 cells were highly susceptible to cell-free EBV infection by Akata and P3HR-1 EBV strains. EBV nuclear antigen (EBNA) and EBV-encoded small RNA (EBER) were detected in the infected cells. Akata and P3HR-1 EBV-infected cell clones were isolated by a limiting dilution technique. The Akata and P3HR-1 EBV-infected clones differed from each other in morphology and growth patterns. Akata EBV-infected clones had lower growth rates than did P3HR-1 EBV-infected clones in both liquid and soft agar mediums. Both the infected HSC-39 cells and the clones expressed EBNA1 and EBER, but did not express EBNA2, latent membrane protein (LMP) 1 and LMP2A. The Q promoter (p), but not the Cp/Wp for EBNA transcription, was active in the infected HSC-39 cells and all clones. No lytic infection was observed in either infected parental cells or any clones. Uninfected HSC-39 cells did not express a principal EBV receptor CD21; however, Akata but not P3HR-1 EBV-infected clones expressed low levels of CD21 mRNA. These results demonstrate that the cellular phenotypes of HSC-39 cells are altered by EBV infection in strain-specific manner. We propose the HSC-39 cell line as a model target for the study of the mechanism and significance of EBV infection in gastric carcinoma. PMID- 15109958 TI - Calcium oscillations in gingival epithelial cells infected with Porphyromonas gingivalis. AB - The periodontal pathogen Porphyromonas gingivalis modulates epithelial cell signal transduction pathways including Ca2+ signaling, and internalizes within the host cell cytoplasm. Since nuclear and cytoplasmic [Ca2+] increases can induce different host cell responses, P. gingivalis-related [Ca2+] changes in these compartments were measured by digital fluorescent imaging microscopy. Non deconvolved and deconvolved fura-2 images showed that P. gingivalis exposure caused human gingival epithelial cells cultured in physiologic [Ca2+] levels to undergo sustained oscillations of [Ca2+] in nuclear and cytoplasmic spaces. However, P. gingivalis invasion was not tightly correlated with intracellular [Ca2+] oscillations, since invasion could significantly precede, or even occur in the absence of, oscillations. [Ca2+] oscillations required a Ca2+ influx, which was completely inhibited by La3+ or 2-APB (2-aminoethoxydiphenyl borate), indicating Ca2+ entry was via a Ca(2+)-permeable channel. Ca2+ entry was likely not via a store-operated channel, since Ca2+ release from intracellular stores was not observed during cellular uptake of P. gingivalis. Hence, uptake of P. gingivalis in gingival epithelial cells induces oscillations in nuclear and cytoplasmic spaces by activating a Ca2+ influx through Ca2+ channels. PMID- 15109959 TI - The antibacterial innate immune response by the mosquito Aedes aegypti is mediated by hemocytes and independent of Gram type and pathogenicity. AB - Previous mosquito studies showed that the hemocyte-mediated innate immune response against Gram- Escherichia coli is phagocytosis, but against Gram+ Micrococcus sp., is melanization. We examined the immune responses mounted by Aedes aegypti towards Gram- Enterobacter cloacae, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Salmonella typhimurium, and Gram+ Bacillus cereus, Enterococcus faecalis, Staphylococcus aureus, and Staphylococcus epidermidis. Using light microscopy, electron microscopy, and survival analysis, this investigation conclusively shows that the factors governing phagocytic vs. melanization responses are complex and independent of bacterial Gram type and pathogenicity. These data provide further evidence that hemocytes are central to the immune response against prokaryotes. PMID- 15109960 TI - Invasion of MDCK epithelial cells with altered expression of Rho GTPases by Trypanosoma cruzi amastigotes and metacyclic trypomastigotes of strains from the two major phylogenetic lineages. AB - In order to invade mammalian cells, Trypanosoma cruzi infective forms cause distinct rearrangements of membrane and host cell cytoskeletal components. Rho GTPases have been shown to regulate three separate signal transduction pathways, linking plasma membrane receptors to the assembly of distinct actin filament structures. Here, we examined the role of Rho GTPases on the interaction between different T. cruzi infective forms of strains from the two major phylogenetic lineages with nonpolarized MDCK cells transfected with different Rho GTPase constructs. We compared the infectivity of amastigotes isolated from infected cells (intracellular amastigotes) with forms generated from the axenic differentiation of trypomastigotes (extracellular amastigotes), and also with metacyclic trypomastigotes. No detectable effect of GTPase expression was observed on metacyclic trypomastigote invasion and parasites of Y and CL (T. cruzi II) strains invaded to similar degrees all MDCK transfectants, and were more infective than either G or Tulahuen (T. cruzi I) strains. Intracellular amastigotes were complement sensitive and showed very low infectivity towards the different transfectants regardless of the parasite strain. Complement-resistant T. cruzi I extracellular amastigotes, especially of the G strain, were more infective than T. cruzi II parasites, particularly for the Rac1V12 constitutively active GTPase transfectant. The fact that in Rac1N17 dominant-negative cells, the invasion of G strain extracellular amastigotes was specifically inhibited suggested an important role for Rac1 in this process. PMID- 15109961 TI - Cathepsin L is crucial for a Th1-type immune response during Leishmania major infection. AB - Prior to the activation of CD4+ T cells, exogenous proteins are digested by endo/lysosomal enzymes in antigen-presenting cells (APCs) to produce antigenic peptides that are presented on MHC class II molecules. In the studies described here, the functional significance of cathepsin L for antigen processing and Th1/Th2 differentiation in experimental leishmaniasis was investigated. We first demonstrated that cathepsin L is one of the candidates for endo/lysosomal enzymes in the processing of soluble Leishmania antigen (SLA) by using CLIK148, a specific inhibitor of cathepsin L. Treatment of BALB/c or DBA/2 mice with CLIK148 exacerbated the disease by enhancing an SLA-specific Th2-type response such as IL 4 production. CLIK148 did not exert any direct influence on Leishmania major promastigotes themselves or on the course of L. major infection in SCID mice. Taken together, these findings suggest that treatment of host mice with CLIK148 affects the processing of SLA in APCs, resulting in the potentiation of Th2-type immune responses and thus leading to exacerbation of the disease. Furthermore, endo/lysosomal cathepsin L was found to be functionally distinct from previously described cathepsins B and D. PMID- 15109962 TI - Schistosoma mansoni: the effect of adrenalectomy on the murine model. AB - Adrenal steroid hormones have been implicated, among others, as one of the most important host factors controlling the onset, establishment, and pathogenesis of schistosomiasis. They appear to inhibit oviposition by Schistosoma mansoni both in vitro and in vivo, and their effect is greatly enhanced when administered in combination with a schistosomicidal drug. Therefore, we hypothesized that adrenalectomy would greatly affect the course of the murine schistosomiasis infection. To test this hypothesis, adrenalectomized mice (Adx) infected with S. mansoni were compared with intact infected and sham-infected controls concerning their mortality rate, numbers of male and female worms, number of eggs, and liver pathology. Compared with controls, Adx infected mice showed an increase of 50% in the mortality rate, as well as 1.7-3 times as many adult worms and twice as many ova per worm pair in their liver. Thus, for the first time, there is evidence that lack of adrenal steroids mediate an increment of the adult worm burden and promote worm fecundity in vivo. The present work was done to test the hypothesis that lack of adrenal steroids enhances adult worm attrition, possibly by their direct effect on the parasite, and by upregulating or downregulating innate and adaptive immune responses. PMID- 15109963 TI - Alternative diphtheria, tetanus and whooping cough immunization schedule to evoke a Th2 tetanus and a Th1 pertussis immune response. AB - In a previous study, using BALB/c mice, we found that while diphtheria (D), tetanus (T) and whooping cough (Pw, whole-cell Bordetella pertussis) immunization induces a Th1/Th2 tetanus response and memory T cells able to proliferate in response to in vitro stimulation with B. pertussis, DTPa immunization induces a Th2 tetanus immune response and no memory T cells that recognize B. pertussis as stimulus. Considering that a pro-inflammatory cytokine production is not necessary for protection against tetanus and therefore should be avoided, an alternative DTP immunization schedule with minimal Pw exposure was assessed in order to obtain a Th2 tetanus response and a Th1 pertussis response. BALB/c mice were primed with DT vaccine at day 0, with Pw vaccine at day 14 and boosted with DTPa vaccine at days 21 and 28. A control group was inoculated with saline. Antibodies against B. pertussis surface antigens, tetanus and diphtheria toxoids were produced by mice. Spleen cells stimulated in vitro with B. pertussis produced IL-6 and IFNgamma. Only IL-5 was produced by cells in response to tetanus toxoid stimulation. These results are in line with the low IgG1/IgG2a ratio for pertussis antibodies compared with those corresponding to tetanus and diphtheria. The immunization protocol presented herein succeeded in producing tetanus and pertussis immune responses of Th2 and Th1 type, respectively. In contrast to previous results obtained with DTPw immunization, no IL-12 production was observed. Our findings provide direct evidence that an immunization protocol with an interval of 14 days between DT and Pw primings, followed by DTPa boosters, can induce appropriate immune responses against DTP vaccine antigens. PMID- 15109964 TI - Construction of recombinant herpes simplex virus type I expressing green fluorescent protein without loss of any viral genes. AB - For use in various applications in research on herpes simplex virus type 1, we attempted to generate recombinant HSV-1 expressing green fluorescent protein (GFP) without any loss of viral genes. Our results were as follows. (i) A recombinant HSV-1 (YK333), in which a GFP expression cassette driven by the Egr-1 promoter was inserted into the intergenic region between UL3 and UL4, was constructed. (ii) YK333 replicated as well as wild-type HSV-1 F strain in Vero cells. (iii) As one application of the recombinant YK333 for research on HSV-1, we developed a system to detect anti-herpetic activity, termed a fluorescence based anti-viral assay. The 50% inhibitory concentration of ganciclovir for YK333 determined using our newly developed assay was comparable to that determined using a plaque reduction assay. YK333 will be a convenient tool for herpes simplex virus research, including such applications as monitoring of viral replication in vitro and in vivo, and rapid screening of potential anti-herpetic agents. PMID- 15109965 TI - Resistance of HIV-infected cells to cytotoxic T lymphocytes. AB - Although cytotoxic T lymphocytes (CTLs) are important for controlling HIV, CTLs are not effective at eradicating HIV infection. Recent studies have revealed a combination of factors that together make HIV-infected cells resistant to CTLs and make anti-HIV CTLs ineffective. These factors likely contribute to prolonged survival of infected target cells, which in turn increases the probability of antigenic variation and immune escape. PMID- 15109966 TI - The role of mhc polymorphism in anti-microbial resistance. AB - The major histocompatibility complex (mhc)-encoded MHC class I and class II molecules present peptide fragments to T cells at every stage of their life (development, survival, persistence and activation). Thereby, these unusually polymorphic molecules critically influence susceptibility to autoimmune and infectious diseases. Here, we examine the mechanistic relationship between mhc polymorphism and anti-microbial resistance/susceptibility. PMID- 15109967 TI - Immunomodulatory action of mycobacterial secretory proteins. AB - The recently discovered RD1 locus encodes proteins that are actively secreted by pathogenic mycobacteria, including Mycobacterium tuberculosis. Since they are missing in non-tuberculous mycobacteria, these proteins are promising not only as candidates for vaccination and diagnostic tests, but also in understanding mycobacterial evasion of protective immunity in susceptible individuals. Here we analyze the possible role of M. tuberculosis secretory proteins in immunity against tuberculosis, with emphasis on their immunomodulatory action and the potential involvement in mycobacterial subversion of the host immune defense. PMID- 15109968 TI - Interaction between histamine-induced itch and experimental muscle pain. AB - Itch sensation can be inhibited by simultaneously applied cutaneous pain at the same skin site via a central mechanism. Deep muscle pain is often associated with sensory changes in the corresponding dermatome. We investigated whether experimentally induced muscle pain has any influence on histamine-induced itch and vice versa in a double blind placebo-controlled study. Experiments were performed in 18 healthy subjects. In nine individuals control iontophoresis of histamine into the forearm produced a distinct itch sensation. Another nine individuals participated in an additional experiment in which histamine and saline were iontophoresed on the forearm in a randomized double-blinded two-way crossover design after intramuscular injection of capsaicin into the ipsilateral brachioradial muscle. Capsaicin-induced muscle pain reduced itch sensation significantly. In contrast, capsaicin-induced muscle pain increased significantly after cutaneous histamine application compared to muscle pain after iontophoresis of saline (placebo). These novel data indicate that muscle pain inhibits itch and histamine increases muscle pain. A bi-directional interaction between cutaneous histamine-sensitive afferents and nociceptive muscle afferents via central mechanisms is suggested. PMID- 15109969 TI - Do pain problems in young school children persist into early adulthood? A 13-year follow-up. AB - DESIGN: In a longitudinal study, 335 children ages 8, 11 and 14, first studied in 1989 were followed-up on two occasions in 1991 and 2002. The subjects filled in questionnaires on pain, the first two times in school, the last as a postal survey. PURPOSES: To determine if headache and back pain during the school years were transitory or if they grew into pain problems in adulthood; to determine predictors of pain. RESULTS: In the 2002 study, 59% of the women and 39% of the men reported pain at 21, 24 and 27 years. A total of 68 (52 women, 16 men) or 20% of the subjects reported pain symptoms in all three studies. The cumulative incidence rate for the presence of pain in the cohort studied was 31% for 1989 2002 and 43% for 1991-2002. Four of the 10 individuals with pain also reported signs of stress. Three predictors were found: reported back pain in 8-14-year olds (p < 0.0001); reported headaches once a week or more in the same age group (p < 0.0001); and a positive response in the ages 10-16 to the question: "Do you often feel nervous?" (OR=2.1, 95% CI 1.3-3.4). When adjusted for age, sex and all psychosocial risk determinants studied in multiple logistic regression, a positive answer to this question was a significant predictor of pain in young adulthood. A positive response by the 10-16-year-olds to "Do you find it difficult to describe your feelings?" was a predictor of pathological anxiety in early adulthood, but stress perceived in childhood/adolescence did not predict future pain or stress. CONCLUSIONS: Since pain reports in childhood and early adolescence seem to be associated with the report of pain in early adulthood, more attention should be given to the way ill-health is managed in adolescence in this vulnerable group. PMID- 15109970 TI - Are changes in fear-avoidance beliefs, catastrophizing, and appraisals of control, predictive of changes in chronic low back pain and disability? AB - Interventions for chronic low back pain (CLBP) often attempt to modify patients' levels of catastrophizing, their fear-avoidance beliefs, and their appraisals of control. Presumably, these interventions are based on the notion that changes in these cognitive factors are related to changes in measures of adjustment. The aim of the present study was to explore whether changes on these cognitive factors were related to changes in CLBP and disability. Fifty-four CLBP patients completed a series of self-report measures prior to beginning a cognitive behavioral based intervention and again upon discharge. Change scores (post treatment score minus pre-treatment score) were calculated for each of the self report measures. The study found that changes in the cognitive factors were not significantly associated with changes in pain intensity. In contrast, reductions in fear-avoidance beliefs about work and physical activity, as well as increased perceptions of control over pain were uniquely related to reductions in disability, even after controlling for reductions in pain intensity, age and sex. The final model explained 71% of the variance in reductions in disability. PMID- 15109971 TI - Catastrophizing and internal pain control as mediators of outcome in the multidisciplinary treatment of chronic low back pain. AB - The aim of the present study was to examine (a) whether a cognitive-behavioral treatment (differentially) affects pain coping and cognition; and (b) whether changes in pain coping and cognition during treatment mediate treatment outcome. Participants in this randomized clinical trial were 148 patients with chronic low back pain attending a multidisciplinary treatment program consisting of operant behavioral treatment plus cognitive coping skills training (N = 59) or group discussion (N = 58) or allocated to a waiting list control condition (N = 31). Patients improved with respect to level of depression, pain behavior and activity tolerance at posttreatment and 12-month follow-up. Treatment also resulted in a short- and long-term decrease in catastrophizing and an enhancement of internal pain control. Changes in catastrophizing and to a lesser degree in internal pain control mediated the reduction in level of depression and pain behavior following treatment. The use of behavioral and cognitive interventions aimed at decreasing catastrophizing thoughts about the consequences of pain and promoting internal expectations of pain control possibly constitute an important avenue of change irrespective of the type of treatment. PMID- 15109972 TI - A polymorphic locus in the intron 16 of the human angiotensin-converting enzyme (ACE) gene is not correlated with complex regional pain syndrome I (CRPS I). AB - Exaggerated neurogenic inflammation has been recognized to be one reason for many CRPS symptoms. Since angiotensin-converting enzyme (ACE) is a key enzyme for the termination of neurogenic inflammation, it has been selected as a candidate gene for CRPS predisposition. A previous report of an insertion/deletion (I/D) polymorphism in intron 16 within the ACE gene implicated an increased risk to develop CRPS I associated with the D allele. However, in the present study the D allele frequency was not increased in CRPS I cases (0.51 for D allele, 0.49 for I allele). Furthermore, there was no co-segregation of any genotype (DD, ID, II) with the CRPS phenotype in 12 selected familial CRPS I cases from six CRPS I families. In conclusion, the results presented herein render this particular ACE gene polymorphism unlikely to be a predisposing factor for CRPS I. PMID- 15109973 TI - Impaired disengagement from threatening cues of impending pain in a crossmodal cueing paradigm. AB - This paper reports an experimental investigation of attentional engagement to and disengagement from cues of impending pain. Pain-free volunteers performed a cueing task in which they were instructed to detect somatosensory and tone targets. Target stimuli were preceded by visual cues informing participants of the modality of the impending stimuli. Participants were randomly assigned to a pain group (n = 54) or to a control group (n = 53). Somatosensory targets consisted of painful electrocutaneous stimuli in the pain group and non-painful vibrotactile targets in the control group. Analyses revealed a similar amount of attentional engagement to both cues signalling somatosensory targets, irrespective of their threat value. However, participants had significantly more difficulty in disengaging attention from a threatening cue of impending pain compared to a cue signalling the non-painful vibrotactile target. Our findings provide further evidence that pain cues demand attention, particularly resulting in impaired disengagement. PMID- 15109974 TI - Mechanically induced axon reflex and hyperalgesia in human UV-B burn are reduced by systemic lidocaine. AB - The mechanisms for the induction of primary mechanical hyperalgesia are unclear. We analyzed the neurogenic axon reflex erythema (flare) following phasic mechanical stimulation in normal and in UV-B irradiated skin. In a cross-over double blind design (n = 10), low dose of systemic lidocaine suppressed mechanical hyperalgesia in sunburned skin and in the mechanically induced flare. Phasic mechanical stimulation, even at painful intensities, did not evoke a flare reaction in normal skin. However, stimulation within the UV-B burn dose dependently provoked an immediate flare reaction. Systemic lidocaine suppressed the mechanically induced flare as well as the mechanical hyperalgesia in sunburned skin, while leaving the impact-induced ratings in normal skin unchanged. Systemic lidocaine reduced these effects of sensitization, but did not reduce ratings in normal skin. As mechanically insensitive ("sleeping") nociceptors have been shown to mediate the axon-reflex in human skin, sensitization of this class of nociceptors might contribute also to the UV-B induced primary mechanical hyperalgesia. PMID- 15109975 TI - Differential release of neurotransmitters from superficial and deep layers of the dorsal horn in response to acute noxious stimulation and inflammation of the rat paw. AB - Experimental evidence suggests that release of neurotransmitters in response to acute noxious stimulation and inflammation can differ in superficial and deeper dorsal horn (DH) laminae. Using two different microdialysis probes, we studied changes in levels of glutamate, aspartate, arginine and GABA in dialysates collected from the surface of the spinal cord and within the DH induced by pinching the paw or paw inflammation. In penthotal anaesthetized rats, a flexible microdialysis probe was placed on the dorsal surface of the L4-L5 or L6-S2 spinal segments. In other rats, a rigid microdialysis probe was implanted within the DH of the same segments. Samples were collected every minute before, during and after pinching the hind paw (acute pain), and every half an hour after injecting either carrageenan or saline into the same paw (inflammation-induced pain). Amino acids were measured by capillary zone electrophoresis with laser-induced fluorescence detection (CZE-LIFD). Pinching the paw induced a significant but short lasting increase in extracellular glutamate and aspartate in dialysates from the surface of the DH. Carrageenan, but not saline, injected into the paw significantly increased concentrations of glutamate, aspartate and arginine both on the surface and within the DH of L4-L5 and also within the DH of the L6-S2 segments. The GABA level was significantly increased following carrageenan only within the DH. The maximum increase on the surface was detected 60-120 min after the onset of inflammation whereas the response within the DH reached a maximum between 150 and 180 min after carrageenan. These results indicate that unlike acute mechanical noxious stimulation which enhances amino acid neurotransmitters in surface dialysate, inflammation induced neurotransmitter release in all layers of the DH suggesting sensitization of the DH. PMID- 15109976 TI - Role of spinal 5-HT(1A) receptors in morphine analgesia and tolerance in rats. AB - We here studied the involvement of spinally located 5-HT(1A) and opioid receptors, in the paradoxical effects that their activation can produce on nociception. Intrathecal (i.t.) injection of the 5-HT(1A) receptor agonist 8 hydroxy-2-[di-n-propylamino] tetralin (8-OH-DPAT) (1-10 microg) induced analgesic effects in the formalin model of tonic pain whereas in the paw pressure test, it decreased the vocalization threshold. In this latter test, i.t. 8-OH-DPAT also markedly reduced the analgesic effect of systemic morphine (5-10 mg/kg, s.c.). At 10 microg, 8-OH-DPAT totally abolished the effect of 5 mg/kg of morphine; this inhibitory effect was antagonized by pre-treatment with 0.63 mg/kg of the 5 HT(1A) antagonist WAY-100635 (N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]-ethyl]-N (2-pyridinyl)-cyclohexanecarboxamide-trihydrochloride). In contrast, the i.t. injection of WAY-100635 (1-10 microg) dose-dependently potentiated the antinociceptive activity of a dose of morphine (2.5 mg/kg, s.c.). Furthermore, WAY-100635 (10 microg, i.t.) potentiated morphine analgesia in morphine-tolerant rats. These findings demonstrate that 5-HT(1A) receptor agonists can act in the spinal cord to produce both hyper- and hypo-algesic effects and play a major role in the opioid analgesia and tolerance. PMID- 15109977 TI - Pharmacological consequences of long-term morphine treatment in patients with cancer and chronic non-malignant pain. AB - BACKGROUND: In patients with pain of malignant origin morphine may be administered in high and often increasing doses during extended periods of time. In patients with chronic pain of non-malignant origin morphine may be an important remedy, and in these cases the goal is to keep the morphine dose stable. The pharmacokinetic as well as the pharmacodynamic consequences of long term morphine treatment with special reference to the two most important metabolites of morphine morphine-6-glucuronide (M-6-G) and morphine-3-glucuronide (M-3-G) remain to be settled. METHODS: Assessments for pain, sedation and other morphine induced side effects were made several times for 19 cancer patients treated with changing doses of oral sustained release (SR) morphine and twice for 17 non-cancer patients treated with stable doses of SR morphine. Blood samples were obtained simultaneously and analysed for contents of morphine, M-3-G and M-6 G by high-performance liquid chromatography (HPLC). RESULTS: Significant correlations were found between the daily dose of SR morphine and plasma morphine (r = 0.469, p < 0.01), plasma M-6-G (r = 0.677, p < 0.01), and plasma M-3-G ((r = 0.827, p < 0.01), in the cancer patient group, but only between the daily dose of SR morphine and plasma M-3-G (0.662, p < 0.01) and plasma M-6-G (0.571, p < 0.01) in the non-cancer patient group. Normalised M-3-G/M and M-6-G/M ratios for the cancer patient group were independent of duration of treatment and daily dose of SR morphine. Likewise in the non-cancer patient group duration of treatment did not influence the metabolite ratios. Correlations between pain score and plasma morphine, M-6-G and M-6-G/M were weak in the cancer patient as well as in the non cancer patient group making it impossible to draw any conclusion regarding the potential contributory analgesic effect of M-6-G. Dryness of the mouth was the most frequent adverse effect reported in the non-cancer as well as the cancer patient group. In the latter group patients complaining of dryness of the mouth had significantly higher plasma morphine and M-6-G concentrations than patients who did not suffer from this side effect. This difference persisted (or was close to significance) when excluding patients receiving antidepressants. CONCLUSION: In the cancer patient group neither dose nor treatment period seems to influence morphine glucuronidation. Likewise in the non-cancer patient group receiving stable doses of morphine duration of treatment does not seem to influence morphine glucuronidation. Dryness of the mouth was positively correlated to high plasma concentrations of morphine and M-6-G. PMID- 15109978 TI - The Fear of Pain questionnaire: factor structure in samples of young, middle-aged and elderly European people. AB - The present study examined the factor structure of the Fear of Pain questionnaire in three independent samples composed of European young adults, middle-aged, and elderly people. Seven hundred and thirty one adults (426 females and 305 males) were presented with the French adaptation of the questionnaire (30 items) and with an Exposure to Painful Situations questionnaire that contained the same items as the Fear of Pain questionnaire and where participants were instructed to indicate all the painful situations they have experienced in the past. When tested on the whole set of 30 items, the correlated three-factor model evidenced in previous studies - Severe pain, Minor pain and Medical pain - poorly fit the data. When tested on a set of 15 two-item parcels, the fit of this model was much better but the correlations between factors were very high. When tested on a reduced set of 15 items, the model fit the data as well as when it was tested on the set of 15 parcels, and the correlations between the three factors were lower. The study also examined the link between previous exposure to pain and fear of pain. The hypothesis that previous "natural" exposure to pain should generally result in a decrease in fear of pain was supported by the data. For 14 items, the exposure effect was moderate to strong. PMID- 15109979 TI - Involvement of protein kinases on the process of erythrophagocytis by Entamoeba histolytica. AB - Erythrophagocytic capacity of trophozoites of Entamoeba histolytica is considered a factor in the virulence of this pathogenic protozoan. We present evidence showing that such activity resembles the ingestion of microorganisms by highly differentiated phagocytic cells, such as macrophages. Previous treatment of the trophozoites with genistein or tyrphostin, inhibitors of tyrosine protein kinases, with staurosporine, a protein kinase C inhibitor, and wortmannin, a fungal metabolite that inhibits phosphoinositide 3-OH kinase, significantly inhibited their erythrophagocytic capacity. PMID- 15109980 TI - Morphometry of nuclear and nucleolar structures in a CaCo-2 cell line. AB - The number of the nucleoli in a CaCo-2 cell nucleus does not generally depend on the quantity of DNA in the nucleus, but nucleolar DNA content is directly proportional to total nuclear DNA. However, in multinucleolar cells (three or more nucleoli), the nucleolar DNA content increases after 96 h incubation in culture without concomitant quantitative changes in nuclear DNA. The percentage of multinucleolar cells and the average number of nucleoli per nucleus increase with increasing incubation time. After 72 and 96 h in culture, multinucleolar cells show distinctive morphologies. The ratio of the sum of nucleolar perimeters to the nuclear perimeter increases linearly when the number of nucleoli in a nucleus increases, but there is no concomitant increase in total nucleolar area or DNA content, except in the 72 and 96 h populations. When the number of nucleoli in CaCo-2 cells increases after 48 and 60 h in culture, the amount of DNA per nucleolus decreases. PMID- 15109981 TI - Effects of ascorbic acid and ascorbic acid 2-phosphate, a long-acting vitamin C derivative, on the proliferation and differentiation of human osteoblast-like cells. AB - In order to investigate the effect of ascorbic acid (AsA) and ascorbic acid 2 phosphate (Asc 2-P), a long-acting vitamin C derivative, on the growth and differentiation of human osteoblast-like cells, we supplemented the culture medium of MG-63 cells with various concentrations (0.25 to 1 mM) of these factors. Asc 2-P significantly stimulated nascent cell growth at all concentrations in the presence of fetal bovine serum (FBS). On the other hand, AsA showed a growth repressive effect depending on its concentration, and that of FBS. Asc 2-P also increased expression of osteoblast differentiation markers, such as collagen synthesis and alkaline phosphatase (ALP) activity. These stimulative activities of Asc 2-P were attenuated by inhibitors of collagen synthesis, indicating that these effects were dependent on collagen synthesis. Electron micrographs of the cells showed the formation of a three-dimensional tissue-like structure endowed with a mature extracellular matrix in the presence of Asc 2-P. PMID- 15109982 TI - Modification of intracellular free calcium in cultured F2408 embryo fibroblasts by 3-substituted-2-thiohydantoin derivatives. AB - 3-substituted-2-thiohydantoin derivatives were synthesized and their structures elucidated by IR, 1H-NMR spectroscopy and elemental analysis. The cytotoxicity of the 2-thiohydantoin derivatives to rat embryo fibroblasts (F2408) in vitro was determined, and the effects of these compounds on intracellular free Ca2+, [Ca2+]i, were measured by spectrofluorophotometry. Cytotoxicity was determined by metabolic reduction of a tetrazolium salt to a formazan dye (MTT assay). Compounds 4 and 7 showed cytotoxic activity, with IC50 values in the range of 1 1.2 microM. Introduction of either chlorophenyl, metoxyphenyl, nitrophenyl or benzyl groups at C-3 resulted in concentration-dependent cytotoxic effects. Compounds 1-6 at 1 microM or more significantly increased [Ca2+]i in a dose dependent manner in the cultured fibroblasts. This action may have been mediated through intracellular calcium stores. PMID- 15109983 TI - Differentiation of chromaffin cells elicited by ELF MF modifies gene expression pattern. AB - Chromaffin cells exposed to extremely low frequency magnetic fields (ELF MF, 60 Hz, 0.7 mT) differentiate into sympathetic neuron-like cells. This complex process must involve both qualitative and quantitative variations in gene expression. This study looks at whether ELF MF treatment provokes changes in the global transcription profile of chromaffin cells, using the RT-Differential Display method. When the gene expression patterns of experimental groups (nerve growth factor (NGF) and ELF MF) were compared to those receiving no treatment, at least 53 transcripts showing differential expression were detected. Eight RT-PCR products, corresponding to six genes, were re-amplified, sequenced and compared with the rat gene bank. Sequence analysis showed that these genes most likely encode: phosphoglucomutase-1, neurofibromatosis-2 interacting protein, microtubule associated protein-2, thiamine pyrophosphokinase, and two unidentified hypothetical proteins (RNOR02022103 and ROR01044577), and that the presumed regulatory regions of these genes contained CTCT-clusters, which are thought to be required for electromagnetic field-dependent gene expression. PMID- 15109984 TI - Differential effects of growth factors and cytokines on the synthesis of SPARC, DNA, fibronectin and alkaline phosphatase activity in human periodontal ligament cells. AB - Growth factors and cytokines play an important role in tissue development and repair. However, it remains unknown how they act on proliferation and differentiation of periodontal ligament cells. In this study, we investigated the effects of several growth factors and cytokines on the synthesis of DNA, alkaline phosphatase (ALPase), fibronectin, and secreted protein acidic and rich in cysteine (SPARC) in human periodontal ligament (HPL) cells. Transforming growth factor-beta (TGF-beta) increased the synthesis of DNA, fibronectin and SPARC, whereas it decreased ALPase activity. Basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF) and tumor necrosis factor-alpha (TNF-alpha) decreased SPARC and ALPase levels, whereas these peptides increased DNA synthesis and did not affect fibronectin synthesis. Epidermal growth factor (EGF) up regulated the synthesis of DNA and fibronectin and inhibited SPARC and ALPase levels. Interleukin-1beta (IL-1beta) decreased the synthesis of DNA, ALPase, fibronectin and SPARC. These findings demonstrate that TGF-beta, bFGF, EGF, PDGF, TNF-alpha and IL-1beta have characteristically different patterns of action on DNA, SPARC, fibronectin and ALPase synthesis by HPL cells. The differences in regulation of function of periodontal ligament cells by these peptides may be involved in the regeneration and repair of periodontal tissue. PMID- 15109985 TI - Effects of triethylene tetraamine on telomerase activity and proliferation in HeLa cells. AB - Telomerase, which is required to maintain telomeres, has attracted considerable attention as a target for anticancer therapy. In this study, we investigated the inhibition of HeLa cell telomerase activity and cell cycle progression by triethylene tetraamine (TETA), using a modified telomeric repeat amplification protocol (TRAP) assay, and flow cytometry. TETA inhibited telomerase activity in HeLa cell extracts, with an IC50 of about 7.8 microM. Coupled with this inhibition, TETA also increased the proportion of cells in the G1 phase of the cell cycle in a dose-dependent manner. These findings demonstrate that TETA is a potent inhibitor of telomerase in micromolar concentrations, and inhibits the proliferation of HeLa cells by arresting them in G1. PMID- 15109986 TI - Backward chromosome movement in crane-fly spermatocytes after UV microbeam irradiation of the interzone and a kinetochore. AB - Single anaphase chromosomes (in crane-fly spermatocytes) moved backwards after double irradiations with an ultraviolet light (UV) microbeam, first of the interzone and then of a kinetochore: the chromosome irradiated at the kinetochore moved backwards rapidly, across the equator and into the other half-spindle. High irradiation doses at the kinetochore were required to induce backward movement. Single irradiations of kinetochores or interzones were ineffective in inducing backward movements. PMID- 15109987 TI - Shape transformation and cytoskeletal reorganization in activated non-mammalian thrombocytes. AB - The nucleated thrombocytes of non-mammalian vertebrates are partially flattened, ovoid cells morphologically distinct from mammalian platelets, and the extent of their functional equivalence is unknown. To test whether they resemble platelets in having similar F-actin-based post-activation stages, rapid fixation/extraction/labeling methods were developed to reveal cytoskeletal organization in dogfish thrombocytes by confocal microscopy. Unactivated cells contained cortical F-actin plus denser F-actin co-localizing with outer marginal band (MB) microtubules. In the post-activation sequence, determined for the first time by continuous observation of individual thrombocytes following thrombin perfusion, cells rounded and blebbed, spread, and eventually flattened extensively. The MB twisted and then became disorganized, with microtubule bundles remaining centrally located and associated with nuclear clefts. In contrast, F-actin occupied blebs and outward-spreading cytoplasm, initially in spiky projections, then predominantly in stress fibers, and inhibitors of F-actin assembly or myosin ATPase blocked shape changes. Thus, the post-activation stages and cytoskeletal events observed in nucleated thrombocytes were found to parallel those of platelets. PMID- 15109988 TI - Small cooperative activity of old rat hepatocytes may depend on composition of the intercellular medium. AB - Ultradian oscillations of protein synthesis were used as a marker of hepatocyte synchronous cooperative activity producing a common rhythm in vitro; amplitude of the rhythm defines expression of the cell cooperation. Dense synchronous and sparse non-synchronous rat hepatocyte cultures on slides in a serum-free incubation medium 199 supplemented with 0.2 mg/ml albumin and 0.5 microg/ml insulin have been studied. The amplitude of the rhythm averaged approximately 2x in dense cultures of young (3 month old) rats than in old (2 year old) rats. But some cultures of young rats had the amplitude patterns similar to cultures of old rats, and vice versa. Addition to the medium of either 0.3 microM bovine brain gangliosides or 2 microM phenylephrine resulted in increase of the oscillation amplitude in dense cultures of old rats to the level inherent in young ones. Addition to the medium of 10% rat blood serum in non-synchronous sparse cultures from young rats resulted in detection of a protein synthetic rhythm. Although after serum from young rats, the rhythm expression was high, the rhythm after serum from old rats had been given was weak. Addition of gangliosides to old-rat serum resulted in synchronization of sparse cultures with amplitudes inherent of young-rat serum. The data tend to the conclusion that cell cooperation depends to a greater extent on the composition of the medium rather than on the age of the cell or animal. PMID- 15109989 TI - Spatiotemporal pattern of calmodulin and [Ca2+]i is related to resumption of meiosis in mouse oocytes. AB - During meiotic maturation, mammalian oocytes undergo a series of morphological and physiological changes that prepare them for fertilization. Calcium-initiated signaling is thought to trigger these processes. In this study, we examine the spatio-temporal pattern of calcium and calmodulin (CaM), its downstream receptor, in order to investigate their association with meiotic maturation. Intracellular free calcium and activated CaM levels were measured using the fluorescent probes Calcium Green-1 and TA-CaM, respectively. The distribution patterns were examined using confocal microscopy. Both calcium and activated CaM showed a dynamic spatiotemporal distribution during meiotic maturation. After release from IBMX buffer, calcium was found to periodically translocate from the perinuclear region to the germinal vesicle (GV) in 90 s intervals. After 90 min, calcium stopped oscillating and became concentrated within the GV. After a further 60 min, the GV broke down and calcium dispersed into the ooplasm, but calcium levels were slightly lower here than in the original nuclear region. Activated CaM also showed a dynamic patterning process similar to calcium. Taking the data from calcium chelation and CaM inhibition together, our results suggest that the dynamic distribution patterns of calcium and activated CaM are crucial for oocyte maturation. PMID- 15109990 TI - GDNF augments survival and differentiation of TH-positive neurons in neural progenitor cells. AB - GDNF plays an important role in the survival and differentiation of primary dopaminergic neurons, but it requires multiple factors for its entire range of activities. This study investigated the effects of GDNF and its cofactors on the development of bFGF-responsive neural progenitor cells (NPCs), mesencephalic and cortical progenitor cells (MP and CP). Various factors were found to have significant inductive effects on the survival and maintenance of these cells in late developmental stages. MP had greater potential than CP to differentiate into dopaminergic neurons. Treatment of NPCs with GDNF and its cofactors enhanced MAP 2 and TH expression, particularly the latter. These findings suggest that NPCs, particularly MP, could develop into more specific neurons if the appropriate factors were applied during the final cell fate specification. They might thus become beneficial sources of donor cells in the treatment of neurological disorders. PMID- 15109991 TI - Signal transmission forces at the cell membrane under debate. PMID- 15109992 TI - Mapping of verbal working memory in nonfluent Chinese-English bilinguals with functional MRI. AB - Existing cognitive and neural imaging studies have suggested a frontoparietal network of multiple, cooperative components for verbal working memory (WM). We used functional MRI to investigate whether this neural network is also involved in the processing of second language by nonfluent bilinguals. Twelve (five males, seven females) native Chinese speakers who had limited English proficiency were scanned while performing working memory tasks in Chinese and English. They were asked to make judgment continuously whether the word presented on the screen was semantically related to (i.e., the semantic tasks) another word presented two words earlier. On a different task (i.e., the phonological tasks), they were asked to make judgment whether the target word rhymed with the other word. A naming and judgment task in each language was adopted to control for the visual process, initial lexical process, and motor responses. Behavioral data showed that subjects performed better at tasks in their native language (Chinese, L1) than in English (L2). Imaging results showed that all working memory tasks in both L1 and L2 elicited a very similar pattern of left-hemisphere-dominated activation in the dorsolateral prefrontal cortex, pars opercularis region, pars triangularis region, precentral cortex, and parietal lobule. Consistent with the behavioral data, the volume of activation in the left opercularis region, left parietal lobule, and right precentral region was greater for L2 than for L1. These results suggest that working memory in L1 and L2 is mediated by a unitary neural system (i.e., frontoparietal region), which is capable of recruiting surrounding cortical resources to meet the increased computational demand caused by low L2 proficiency. PMID- 15109993 TI - Sentence complexity and input modality effects in sentence comprehension: an fMRI study. AB - Cortical regions engaged by sentence processing were mapped using functional MRI. The influence of input modality (spoken word vs. print input) and parsing difficulty (sentences containing subject-relative vs. object-relative clauses) was assessed. Auditory presentation was associated with pronounced activity at primary auditory cortex and across the superior temporal gyrus bilaterally. Printed sentences by contrast evoked major activity at several posterior sites in the left hemisphere, including the angular gyrus, supramarginal gyrus, and the fusiform gyrus in the occipitotemporal region. In addition, modality-independent regions were isolated, with greatest overlap seen in the inferior frontal gyrus (IFG). With respect to sentence complexity, object-relative sentences evoked heightened responses in comparison to subject-relative sentences at several left hemisphere sites, including IFG, the middle/superior temporal gyrus, and the angular gyrus. These sites showing modulation of activity as a function of sentence type, independent of input mode, arguably form the core of a cortical system essential to sentence parsing. PMID- 15109994 TI - GABA A receptor abnormalities in Prader-Willi syndrome assessed with positron emission tomography and [11C]flumazenil. AB - Prader-Willi syndrome (PWS) is a multi-system disorder characterized clinically by abnormal mental and physical development. PWS patients have a deletion in an imprinted region on paternal chromosome 15 (15q11-13), maternal disomy for this segment, or rarely, a chromosomal imprinting center deletion that gives rise to suppression of the equivalent paternal genes. Within the affected segment of chromosome 15 are genes encoding the alpha(5), beta(3) and gamma(3) subunits of the gamma-aminobutyric acid type-A (GABA(A)) receptor. Therefore, altered neurobehavioral function could arise in PWS due directly to altered GABA(A) receptor composition and expression, or alternatively, from brain developmental and maturational effects of these or other genes in the imprinted region. The aim of the present study was to assess cerebral GABA(A) receptors in PWS with the use of positron emission tomography of the benzodiazepine binding site employing [11C]flumazenil ([11C]FMZ). A reduction in [11C]FMZ binding was found predominantly in the cingulate, frontal and temporal neocortices and insula in six adult PWS patients compared to nine normal subjects. A possible role for the deleted beta(3) subunit gene in PWS is supported in part by the wide cortical distribution of its mRNA expression and the effects of experimental knockouts on benzodiazepine binding described in prior studies. Altered GABA(A) receptor composition or number in these cortical regions may account for neurobehavioral abnormalities in PWS including mild mental retardation, poor impulse control, and impaired responses to somatic pain. PMID- 15109995 TI - Overt propositional speech in chronic nonfluent aphasia studied with the dynamic susceptibility contrast fMRI method. AB - This study examined activation levels in the left (L) supplementary motor area (SMA) and the right (R) SMA (separately), and activation in nine R perisylvian language homologues during overt, propositional speech in chronic nonfluent aphasia patients. Previous functional imaging studies with a variety of chronic aphasia patients have reported activation in these regions during different language tasks, however, overt propositional speech has not been examined. In the present research, four nonfluent aphasia patients were studied during overt elicited propositional speech at 4-9 years post-single L hemisphere stroke, which spared the SMA. The dynamic susceptibility contrast (DSC) method of functional MRI was used to calculate relative cerebral blood volume (relCBV) for cortical regions of interest (ROIs) during the first-pass bolus of gadolinium during two conditions: (1) pattern (silent viewing of checkerboard patterns) and (2) story (overt, elicited propositional speech describing sequential pictures, which formed a story). During the story condition, controls had significantly higher relCBV in L SMA than in R SMA; aphasics, however, had significantly higher relCBV in R SMA than in L SMA. During the pattern condition, no significant differences were observed between the L SMA and the R SMA for either controls or aphasics. In addition, aphasics had significantly higher relCBV in the R sensorimotor mouth during story than pattern. This R sensorimotor mouth relCBV was also significantly higher in aphasics than controls during story, and the two groups did not differ during pattern. The overall mean relCBV for the nine R perisylvian ROIs was significantly higher for aphasics than controls during both story and pattern. These results suggest that poor modulation, including possible over activation of R sensorimotor mouth and other R perisylvian language homologues may underlie in part, the hesitant, poorly articulated, agrammatic speech associated with nonfluent aphasia. PMID- 15109996 TI - Analysis of neural mechanisms underlying verbal fluency in cytoarchitectonically defined stereotaxic space--the roles of Brodmann areas 44 and 45. AB - We investigated neural activations underlying a verbal fluency task and cytoarchitectonic probabilistic maps of Broca's speech region (Brodmann's areas 44 and 45). To do so, we reanalyzed data from a previous functional magnetic resonance imaging (fMRI) [Brain 125 (2002) 1024] and from a cytoarchitectonic study [J. Comp. Neurol. 412 (1999) 319] and developed a method to combine both data sets. In the fMRI experiment, verbal fluency was investigated in 11 healthy volunteers, who covertly produced words from predefined categories. A factorial design was used with factors verbal class (semantic vs. overlearned fluency) and switching between categories (no vs. yes). fMRI data analysis employed SPM99 (Statistical Parametric Mapping). Cytoarchitectonic maps of areas 44 and 45 were derived from histologic sections of 10 postmortem brains. Both the in vivo fMRI and postmortem MR data were warped to a common reference brain using a new elastic warping tool. Cytoarchitectonic probability maps with stereotaxic information about intersubject variability were calculated for both areas and superimposed on the functional data, which showed the involvement of left hemisphere areas with verbal fluency relative to the baseline. Semantic relative to overlearned fluency showed greater involvement of left area 45 than of 44. Thus, although both areas participate in verbal fluency, they do so differentially. Left area 45 is more involved in semantic aspects of language processing, while area 44 is probably involved in high-level aspects of programming speech production per se. The combination of functional data analysis with a new elastic warping tool and cytoarchitectonic maps opens new perspectives for analyzing the cortical networks involved in language. PMID- 15109997 TI - Mapping distributed sources of cortical rhythms in mild Alzheimer's disease. A multicentric EEG study. AB - The study aimed at mapping (i) the distributed electroencephalographic (EEG) sources specific for mild Alzheimer's disease (AD) compared to vascular dementia (VaD) or normal elderly people (Nold) and (ii) the distributed EEG sources sensitive to the mild AD at different stages of severity. Resting EEG (10-20 electrode montage) was recorded from 48 mild AD, 20 VaD, and 38 Nold subjects. Both AD and VaD patients had 24-17 of mini mental state examination (MMSE). EEG rhythms were delta (2-4 Hz), theta (4-8 Hz), alpha 1 (8-10.5 Hz), alpha 2 (10.5 13 Hz), beta 1 (13-20 Hz), and beta 2 (20-30 Hz). Cortical EEG sources were modeled by low resolution brain electromagnetic tomography (LORETA). Regarding issue i, there was a decline of central, parietal, temporal, and limbic alpha 1 (low alpha) sources specific for mild AD group with respect to Nold and VaD groups. Furthermore, occipital alpha 1 sources showed a strong decline in mild AD compared to VaD group. Finally, distributed theta sources were largely abnormal in VaD but not in mild AD group. Regarding issue ii, there was a lower power of occipital alpha 1 sources in mild AD subgroup having more severe disease. Compared to previous field studies, this was the first investigation that illustrated the power spectrum profiles at the level of cortical (macroregions) EEG sources in mild AD patients having different severity of the disease with respect to VaD and normal subjects. Future studies should evaluate the clinical usefulness of this approach in early differential diagnosis, disease staging, and therapy monitoring. PMID- 15109998 TI - Functional abnormalities in symptomatic concussed athletes: an fMRI study. AB - Our aim was to quantify with functional magnetic resonance imaging (fMRI) changes in brain activity in concussed athletes and compare the results with those of normal control subjects. Regional brain activations associated with a working memory task were obtained from a group of concussed athletes (15 symptomatic, 1 asymptomatic) and eight matched control subjects, using blood oxygen level dependent (BOLD) fMRI. The average percent signal change from baseline to working memory condition in each region of interest was computed. Symptomatic concussed athletes demonstrated task-related activations in some but not all the regions of interest, even when they performed as well as the control subjects. Furthermore, several concussed athletes had additional increases in activity outside the regions of interest, not seen in the control group. Quantitative analysis of BOLD signals within regions of interest revealed that, in general, concussed athletes had different BOLD responses compared to the control subjects. The task-related activation pattern of the one symptom-free athlete was comparable to that of the control group. We also repeated the study in one athlete whose symptoms had resolved. On the first study, when he was still symptomatic, less task-related activations were observed. On follow-up, once his symptoms had disappeared, the task-related activations became comparable to those of the control group. These results demonstrate the potential of fMRI, in conjunction with the working memory task, to identify an underlying pathology in symptomatic concussed individuals with normal structural imaging results. PMID- 15109999 TI - Integration of fMRI and simultaneous EEG: towards a comprehensive understanding of localization and time-course of brain activity in target detection. AB - fMRI and EEG are complimentary methods for the analysis of brain activity since each method has its strength where the other one has limits: The spatial resolution is thus in the range of millimeters with fMRI and the time resolution is in the range of milliseconds with EEG. For a comprehensive understanding of brain activity in target detection, nine healthy subjects (age 24.2 +/- 2.9) were investigated with simultaneous EEG (27 electrodes) and fMRI using an auditory oddball paradigm. As a first step, event-related potentials, measured inside the scanner, have been compared with the potentials recorded in a directly preceding session in front of the scanner. Attenuated amplitudes were found inside the scanner for the earlier N1/P2 component but not for the late P300 component. Second, an independent analysis of the localizations of the fMRI activations and the current source density as revealed by low resolution electromagnetic tomography (LORETA) has been done. Concordant activations were found in most regions, including the temporoparietal junction (TPJ), the supplementary motor area (SMA)/anterior cingulate cortex (ACC), the insula, and the middle frontal gyrus, with a mean Euclidean distance of 16.0 +/- 6.6 mm between the BOLD centers of gravity and the LORETA-maxima. Finally, a time-course analysis based on the current source density maxima was done. It revealed different time-course patterns in the left and right hemisphere with earlier activations in frontal and parietal regions in the right hemisphere. The results suggest that the combination of EEG and fMRI permits an improved understanding of the spatiotemporal dynamics of brain activity. PMID- 15110000 TI - An evaluation of thresholding techniques in fMRI analysis. AB - This paper reviews and compares individual voxel-wise thresholding methods for identifying active voxels in single-subject fMRI datasets. Different error rates are described which may be used to calibrate activation thresholds. We discuss methods which control each of the error rates at a prespecified level alpha, including simple procedures which ignore spatial correlation among the test statistics as well as more elaborate ones which incorporate this correlation information. The operating characteristics of the methods are shown through a simulation study, indicating that the error rate used has an important impact on the sensitivity of the thresholding method, but that accounting for correlation has little impact. Therefore, the simple procedures described work well for thresholding most single-subject fMRI experiments and are recommended. The methods are illustrated with a real bilateral finger tapping experiment. PMID- 15110001 TI - Cytochrome-c-oxidase redox changes during visual stimulation measured by near infrared spectroscopy cannot be explained by a mere cross talk artefact. AB - The detection of redox changes in cytochrome-c-oxidase ([Cyt-ox]) in response to cerebral activation by non-invasive NIRS is hampered by methodological spectroscopic issues related to the modification of the Beer-Lambert law. Also, the question whether a change in the enzyme's redox-state is elicited by functional stimulation is unresolved. In a previous study, we found physiological evidence in favour of an activation-induced increase in oxidation of the enzyme [J. Cereb. Blood Flow Metab. 19 (1999) 592], while in a second study on spectroscopic cross talk, we found that the [Cyt-ox] changes to potentially be an artefact of the spectroscopic approach [J. Biomed. Opt. 7 (2002) 51]. Here, we use two different stimuli which differentially activate areas either rich or poor in [Cyt-ox] content (blob/interblob in visual cortex V1 and pale/thin stripes in V2) to further clarify this apparent discrepancy. In a first experiment, two stimuli were presented in an alternating fashion for 20 s and all stimulation periods were separated by resting periods of 40 s. We observed similar changes in [Cyt-ox] for both stimuli. To become more sensitive to the potentially very small optical changes related to changes in [Cyt-ox], we tried to minimise global haemodynamic and metabolic effects in a second experiment by omitting the resting periods. Our hypothesis was that [Cyt-ox] changes could be fully explained by cross talk as it is predicted from our last study [J. Biomed. Opt. 7 (2002) 51]. However, in more than half of the experiments, we were not able to model the changes in Cyt-ox calculated from measured attenuation spectra as a cross talk artefact. We interpret this finding as an argument in favour of the existence of [Cyt-ox] changes in response to functional stimulation. This finding, however, does not lessen the liability of the [Cyt-ox] changes to cross talk and calls for great caution when [Cyt-ox] changes are derived from NIRS measurements based on the modified Beer-Lambert approach. Further (invasive) validation studies are required. PMID- 15110002 TI - An integrative MEG-fMRI study of the primary somatosensory cortex using cross modal correspondence analysis. AB - We develop a novel approach of cross-modal correspondence analysis (CMCA) to address whether brain activities observed in magnetoencephalography (MEG) and functional magnetic resonance imaging (fMRI) represent a common neuronal subpopulation, and if so, which frequency band obtained by MEG best fits the common brain areas. Fourteen adults were investigated by whole-head MEG using a single equivalent current dipole (ECD) and synthetic aperture magnetometry (SAM) approaches and by fMRI at 1.5 T using linear time-invariant modeling to generate statistical maps. The same somatosensory stimulus sequences consisting of tactile impulses to the right sided: digit 1, digit 4 and lower lip were used in both neuroimaging modalities. To evaluate the reproducibility of MEG and fMRI results, one subject was measured repeatedly. Despite different MEG dipole locations and locations of maximum activation in SAM and fMRI, CMCA revealed a common subpopulation of the primary somatosensory cortex, which displays a clear homuncular organization. MEG activity in the frequency range between 30 and 60 Hz, followed by the ranges of 20-30 and 60-100 Hz, explained best the defined subrepresentation given by both MEG and fMRI. These findings have important implications for improving and understanding of the biophysics underlying both neuroimaging techniques, and for determining the best strategy to combine MEG and fMRI data to study the spatiotemporal nature of brain activity. PMID- 15110003 TI - An automated algorithm for the computation of brain volume change from sequential MRIs using an iterative principal component analysis and its evaluation for the assessment of whole-brain atrophy rates in patients with probable Alzheimer's disease. AB - This article introduces an automated method for the computation of changes in brain volume from sequential magnetic resonance images (MRIs) using an iterative principal component analysis (IPCA) and demonstrates its ability to characterize whole-brain atrophy rates in patients with Alzheimer's disease (AD). The IPCA considers the voxel intensity pairs from coregistered MRIs and identifies those pairs a sufficiently large distance away from the iteratively determined PCA major axis. Analyses of simulated and real MRI data support the underlying assumption of a linear relationship in paired voxel intensities, identify an outlier distance threshold that optimizes the trade-off between sensitivity and specificity in the detection of small volume changes while accounting for global intensity changes, and demonstrate an ability to detect changes as small as 0.04% of brain volume without confounding effects of between-scan shifts in voxel intensity. In eight patients with probable AD and eight age-matched normal control subjects, the IPCA was comparable to the established but partly manual digital subtraction (DS) method in characterizing annual rates of whole-brain atrophy: resulting rates were correlated (Spearman rank correlation = 0.94, P < 0.0005) and comparable in distinguishing probable AD from normal aging (IPCA detected atrophy rates: 2.17 +/- 0.52% per year in the patients vs. 0.41 +/- 0.22% per year in the controls [Wilcoxon-Mann-Whitney test P = 7.8 x 10(-4)]; DS detected atrophy rates: 3.51 +/- 1.31% per year in the patients vs. 0.48 +/- 0.29% per year in the controls [P = 7.8 x 10(-4)]). The IPCA could be used in tracking the progression of AD, evaluating the disease-modifying effects of putative treatments, and investigating the course of other normal and pathological changes in brain morphology. PMID- 15110004 TI - The topography of white matter hyperintensities on brain MRI in healthy 60- to 64 year-old individuals. AB - We report the topography of brain white matter hyperintensities (WMHs) on T2 weighted fluid attenuated inversion recovery (FLAIR) magnetic resonance imaging in 477 healthy subjects aged 60-64 years selected randomly from the community. WMHs were delineated by using a computer algorithm. We found that all subjects had periventricular WMHs and 96.6% subjects also had deep WMHs. The mean volume of WMHs was 4.9 ml, comprising 0.83% of the white matter, of which 1.2 ml was severe in intensity. The deep WMHs were distributed throughout the cerebral hemispheres, with the occipital and frontal white matter bearing the greatest burden. The territory of the lenticulostriate arteries had the greatest WMHs. A white matter region of 4 mm adjacent to the cortex was not affected by hyperintensities. The mean (SD) number of discrete WMHs was 19.6 (7.1) per subject, of which 6.1 (4.4) were severe in intensity. Nearly half (48.6%) of the subjects had at least one large WMH (>12 mm diameter) and one eighth (12.5%) of the subjects had at least one large WMH that appeared to be severe in MRI. The overall load of WMHs was similar in men and women, but the latter had a higher proportion of their white matter so affected. This study provides the first detailed topographic analysis of WMHs in a large representative middle-aged sample, emphasizes their high prevalence in mid-adult life and raises issues about their etiology and significance. PMID- 15110005 TI - Activation in the premotor cortex during mental calculation in patients with Alzheimer's disease: relevance of reduction in posterior cingulate metabolism. AB - In vivo brain imaging of people with Alzheimer's disease (AD) has suggested the presence of functional disintegration in the posterior-anterior brain network from the posterior cingulate cortex (PCC) to the prefrontal cortex. To investigate the relationship between the baseline posteromedial metabolism and prefrontal neural activity during cognitomnemonic tasks in AD patients, we measured both glucose metabolism at baseline and cerebral blood flow (CBF) during the execution of mental calculation tasks (serial number subtraction) in 10 early stage AD patients and six healthy subjects. The present study employed positron emission tomography with (18)F-fluorodeoxyglucose and H(2)(15)O. Group comparison using the region-of-interest (ROI) method and voxel-based statistical parametric mapping (SPM99) showed significant reduction in glucose metabolism in the PCC of the AD group. The PCC metabolism in the AD group was negatively correlated with scores on the Mini-Mental State Examination and with correct responses to the arithmetic task. During the arithmetic task, regional CBF increased significantly in the left parietal and bilateral prefrontal cortices in the normal group, whereas the bilateral premotor cortices were significantly activated in the AD group. Regression analysis showed a significant inverse correlation between the premotor activation and the baseline PCC metabolism in the AD group. These results suggest that the premotor cortex plays a compensatory role in executing mental calculations in AD patients with reduced posteromedial functions, which might reflect the dynamic aspect of the pathophysiology of early AD. PMID- 15110006 TI - Retrieval of abstract semantics. AB - Behavioural and neuropsychological evidence suggests that abstract and concrete concepts might be represented, retrieved and processed differently in the human brain. Using fMRI, we demonstrate that retrieval of abstract relative to sensory based semantics during synonym judgements increased activation in a left frontotemporal system that has been associated with semantic processing particularly at the sentence level. Since activation increases were observed irrespective of the degree of difficulty, we suggest that these differential activations might reflect a particular retrieval mechanism or strategy for abstract concepts. In contrast to sensory-based semantics, the meaning of abstract concepts is largely specified by their usage in language rather than by their relations to the physical world. Subjects might therefore generate an appropriate semantic sentential context to fully explore and specify the meaning of abstract concepts. Our results also explain why abstract semantics is vulnerable to left frontotemporal lesions. PMID- 15110007 TI - The brain network associated with acquiring semantic knowledge. AB - There is ongoing debate about how semantic information is acquired, whether this occurs independently of episodic memory, and what role, if any, brain areas such as hippocampus are required to play. We used auditory stimuli and functional MRI (fMRI) to assess brain activations associated with the incidental acquisition of new and true facts about the world of the sort we are exposed to day to day. A control task was included where subjects heard sentences that described novel scenarios involving unfamiliar people, but these did not convey general knowledge. The incidental encoding task was identical for two stimulus types; both shared the same episodic experience (lying in the brain scanner) and conveyed complex information. Despite this, and considering only those stimuli successfully encoded, compared to a baseline task, a more extensive network of brain regions was found to be associated with exposure to new facts including the hippocampus. Direct comparison between the two stimulus types revealed greater activity in dorsal, ventrolateral and dorsomedial prefrontal cortex, medial dorsal nucleus of the thalamus, and temporal cortex for fact stimuli. The findings suggest that successful encoding is not invariably associated with activation of one particular brain network. Rather, activation patterns may depend on the type of materials being acquired, and the different processes they engender when subjects encode. Qualitatively, from postscan debriefing sessions, it emerged that the factual information was found to be potentially more useful. We suggest that current or prospective utility of incoming information may be one factor that influences the processes engaged during encoding and the concomitant neuronal responses. PMID- 15110008 TI - A dynamic system model-based technique for functional MRI data analysis. AB - Signals in functional magnetic resonance imaging (fMRI) are influenced by physiological fluctuations in addition to local brain activity. We have proposed a dynamic system model-based technique for separation of signal changes related to brain activation inputs from those related to physiological fluctuations. We applied this technique to a visual fMRI experiment to determine the validity and feasibility of this technique for fMRI data analyses. Gradient-echo echo planar images were obtained from 12 healthy volunteers with a Siemens ALLEGRA operating at 3 T, with a repetition time of 500 ms, echo time of 20 ms, field of view of 200-210 mm, matrix size of 64 x 64, and slice thickness of 5 mm. Twelve runs with two stimulation periods of varied duration (2-8 s) with 8-Hz flickering illumination were obtained for each subject. Local signal changes were modeled by an autoregressive model with two exogenous inputs, a visual stimulation input and a global reference signal. Local signal changes were appropriately predicted not only for stimulation periods but also resting periods. A significant linear relationship was found between model static gain based on the dynamic system modeling and beta coefficient based on a general linear model (GLM) analysis for active voxels in the primary visual cortex (analysis of covariance [ANCOVA], P < 0.001; estimated parameter, 0.967; 95% confidence interval, 0.734-1.201). This dynamic system model-based technique is sufficiently accurate and feasible for use in extracting signal changes related to brain activation inputs from measured signals with physiological fluctuations. PMID- 15110009 TI - Voxel-based morphometry of comorbid schizophrenia and learning disability: analyses in normalized and native spaces using parametric and nonparametric statistical methods. AB - We employed voxel-based morphometry (VBM) to compare the distributions of grey matter found in structural magnetic resonance imaging (MRI) brain scans of patients with comorbid learning disability with schizophrenia, schizophrenia alone, learning disability alone, and normal controls. Our primary aim was to replicate a previous region of interest (ROI) finding that comorbids and schizophrenics belong to the same population. Nonparametric analysis in normalized space showed no significant differences in grey matter distribution between the comorbid and schizophrenia groups. Furthermore, this analysis showed significant grey matter reductions in the comorbid and schizophrenia groups when compared to the learning-disabled or the normal controls. Parametric analysis localized the significant grey matter reductions between the normal controls and the comorbid and schizophrenia groups to the prefrontal and temporal lobes. It also identified an area of increased grey matter, on the inferior aspect of the postcentral gyrus, in the learning-disabled alone compared to the other groups. Native space parametric and nonparametric analyses, based on modulation of the normalized scans, confirmed the similarity in grey matter distribution of the comorbid and schizophrenia groups. Results confirm the ROI finding that in native space the learning-disabled group possesses the least and normal controls the most grey matter for the cohort. An increase in the basal ganglia of patients with schizophrenia vs. the learning-disabled, probably attributable to antipsychotic medication, was identified in the native space analysis. The native space results did not however register statistically significant temporal lobe reductions found under normalized analysis between schizophrenics and normal controls. This may be attributable to minor physical anomalies (MPA) in the schizophrenic cranium. Overall, these VBM results replicate previous ROI findings and are compatible with the view that comorbid learning disability with schizophrenia is a severe form of schizophrenia, rather than a consequence of learning disability. VBM has the facility to compare grey matter distributions in this structurally diverse cohort. PMID- 15110010 TI - Optimized 3 T EPI of the amygdalae. AB - The optimum parameters for single-shot gradient-recalled (GR) EPI-based fMRI studies of the limbic region are systematically established at 3 T via their ability to mitigate intravoxel dephasing-measured via SNR and T2* in the amygdalae-and their implications for temporal resolution (or brain coverage). Conventional imaging parameters (64 x 64 matrix size and 4-6 mm thick slices) are confirmed to be inadequate for functional studies at 3 T. Measurements of main magnetic field variations across the amygdalae suggest that such variations are equal in the craniocaudal and anterior-posterior directions, and slightly lower in the mediolateral direction, with this and other considerations leading us to conclude an oblique axial orientation to be most suitable. In-plane resolution of approximately 1.7 mm was sufficient to recover signal in the area of the amygdalae. SNR was found to peak at a slice thickness of between 2.0 and 2.5 mm, dependent on the subject. T2* time in the amygdalae was measured with a standard EPI protocol to be 22 +/- 3 ms. Using the optimized (high resolution) EPI protocol proposed here, the measured T2* time increased to 48 +/- 2 ms (compared with 43 +/- 3 ms for a reference FLASH scan), only slightly lower than the cortex (49 +/- 2 ms measured with optimized EPI and 52 +/- 2 ms with FLASH). The FLASH measurement of 43 ms is taken to be a suitable effective echo time (TE(eff)) to achieve maximum BOLD sensitivity in the amygdalae. Time series data acquired with these parameters showed a 60% increase in SNR in the amygdala over that obtained with a standard low-resolution protocol and suggest sufficient SNR and BOLD sensitivity to make functional studies feasible. Arteries, but no substantial draining veins, were found in high-resolution BOLD venograms of the region. Our results indicate that EPI protocols need to be carefully optimized for structures of interest if reliable results from single subjects are to be established in this brain region. PMID- 15110011 TI - A functional MRI study of the influence of practice on component processes of working memory. AB - Previous neuroimaging studies have shown that neural activity changes with task practice. The types of changes reported have been inconsistent, however, and the neural mechanisms involved remain unclear. In this study, we investigated the influence of practice on different component processes of working memory (WM) using a face WM task. Event-related functional magnetic resonance imaging (fMRI) methodology allowed us to examine signal changes from early to late in the scanning session within different task stages (i.e., encoding, delay, retrieval), as well as to determine the influence of different levels of WM load on neural activity. We found practice-related decreases in fMRI signal and effects of memory load occurring primarily during encoding. This suggests that practice improves encoding efficiency, especially at higher memory loads. The decreases in fMRI signal we observed were not accompanied by improved behavioral performance; in fact, error rate increased for high WM load trials, indicating that practice related changes in activation may occur during a scanning session without behavioral evidence of learning. Our results suggest that practice influences particular component processes of WM differently, and that the efficiency of these processes may not be captured by performance measures alone. PMID- 15110012 TI - BOLD-contrast functional MRI signal changes related to intermittent rhythmic delta activity in EEG during voluntary hyperventilation-simultaneous EEG and fMRI study. AB - Differences in the blood oxygen level dependent (BOLD) signal changes were studied during voluntary hyperventilation (HV) between young healthy volunteer groups, (1) with intermittent rhythmic delta activity (IRDA) (N = 4) and (2) controls (N = 4) with only diffuse arrhythmic slowing in EEG (normal response). Subjects hyperventilated (3 min) during an 8-min functional MRI in a 1.5-T scanner, with simultaneous recording of EEG (successful with N = 3 in both groups) and physiological parameters. IRDA power and average BOLD signal intensities (of selected brain regions) were calculated. Hypocapnia showed a tendency to be slightly lighter in the controls than in the IRDA group. IRDA power increased during the last minute of HV and ended 10-15 s after HV. The BOLD signal decreased in white and gray matter after the onset of HV and returned to the baseline within 2 min after HV. The BOLD signal in gray matter decreased approximately 30% more in subjects with IRDA than in controls, during the first 2 min of HV. This difference disappeared (in three subjects out of four) during IRDA in EEG. BOLD signal changes seem to depict changes, which precede IRDA. IRDA due to HV in healthy volunteers represent a model with a clearly defined EEG pattern and an observable BOLD signal change. PMID- 15110013 TI - Scale invariant adaptation in fusiform face-responsive regions. AB - Several functional neuroimaging studies have observed response adaptation in face sensitive regions when repeating identical face stimuli. To address whether this was due to low-level stimulus properties or facial identity, we decomposed pictures of faces into pictures preserving only the lower or higher parts of the normal frequency spectrum. In an event-related functional neuroimaging study, pairs of such pictures were sequentially presented that showed the same or different persons in the same or different frequency bands. This factorial design allowed to separate effects related to repetition of personal identity from those related to identical stimulus properties. In a random effects group analysis, activation in the right fusiform region was affected by repetition of personal identity regardless of changing or constant spatial scale. Responses in the more medial and posterior fusiform and lingual regions adapted with repetition of the same frequency band. An analysis in regions of interest determined individually as face responsive showed that repetition decreases for the same faces in fusiform face-responsive regions generalized across spatial frequency bands. Our results therefore point to a role of this area in discriminating individual faces at a level of representation that is invariant to changes in low-level stimulus properties, as spatial scale. The same invariance could not be detected in more posterior occipital face-responsive regions. PMID- 15110014 TI - Activity in ventromedial prefrontal cortex covaries with sympathetic skin conductance level: a physiological account of a "default mode" of brain function. AB - We examined neural activity related to modulation of skin conductance level (SCL), an index of sympathetic tone, using functional magnetic resonance imaging (fMRI) while subjects performed biofeedback arousal and relaxation tasks. Neural activity within the ventromedial prefrontal cortex (VMPFC) and the orbitofrontal cortex (OFC) covaried with skin conductance level (SCL), irrespective of task. Activity within striate and extrastriate cortices, anterior cingulate and insular cortices, thalamus, hypothalamus and lateral regions of prefrontal cortex reflected the rate of change in electrodermal activity, highlighting areas supporting transient skin conductance responses (SCRs). Successful performance of either biofeedback task (where SCL changed in the intended direction) was associated with enhanced activity in mid-OFC. The findings point to a dissociation between neural systems controlling basal sympathetic tone (SCL) and transient skin conductance responses (SCRs). The level of activity in VMPFC has been related to a default mode of brain function and our findings provide a physiological account of this state, indicating that activity within VMPFC and OFC reflects a dynamic between exteroceptive and interoceptive deployment of attention. PMID- 15110015 TI - fMRI analysis with the general linear model: removal of latency-induced amplitude bias by incorporation of hemodynamic derivative terms. AB - Functional magnetic resonance imaging (fMRI) data are often analyzed using the general linear model employing a hypothesized neural model convolved with a hemodynamic response function. Mismatches between this hemodynamic model and the data can be induced by spatially varying delays or slice-timing differences. It is common practice to desensitize the analysis to such delays by incorporation of the hemodynamic model plus its temporal derivative. The rationale often used is that additional variance will be captured and regressed out from the data. Though this is true, it ignores the potential for amplitude bias induced by small model mismatches due to, for example, variable hemodynamic delays and is not helpful for "random effects" analyses which typically do not account for the first level variance at all. Amplitude bias is due to the use of only the nonderivative portion of the model in the final test for significant amplitudes. We propose instead testing an amplitude value that is a function of both the nonderivative and the derivative terms of the model. Using simulations, we show that the proposed amplitude test does not suffer from delay-induced bias and that a model incorporating temporal derivatives is a more natural test for amplitude differences. The proposed test is applied in a random-effects analysis of 100 subjects. It reveals increased amplitudes in areas consistent with the task, with the largest increases in regions with greater hemodynamic delays. PMID- 15110016 TI - Assessment of cerebral hemodynamics and oxygen extraction using dynamic susceptibility contrast and spin echo blood oxygenation level-dependent magnetic resonance imaging: applications to carotid stenosis patients. AB - Blood oxygenation level-dependent (BOLD) magnetic resonance imaging (MRI) has been recently used to quantify cerebral blood volume (CBV) and oxygen extraction ratio (OER). In the present study, we have exploited the intravascular BOLD model to assess gray matter (GM) OER at hemispheric level using parenchymal T(2) and CBV data at 1.5 T, obtained by single spin echo and dynamic susceptibility contrast (DSC) perfusion MRI, respectively. An OER of 0.40 +/- 0.07 was determined in gray matter for control subjects. A group of carotid stenosis (CS) patients (n = 22) was examined by multiparametric MRI. The degree of CS was determined by contrast agent-enhanced magnetic resonance angiography. Within the group, eight cases with <70% narrowing of a carotid lumen, nine cases with 70 99%, and five cases with complete occlusion of either carotid arteries were found. DSC MRI revealed abnormalities in 14 patients in dynamic parameters of perfusion images. These included four cases with elevated hemispheric gray matter CBV ipsilateral to the stenosis, above 2 SD of the level determined in control subjects. These four patients showed large variation in the degree of stenosis. We also found three cases with ipsilateral gray matter CBV below 2 SD of the control value, two of these with >70% stenosis. Gray matter OER ipsilateral to the stenosis was above 2 SD of the control range in eight CS patients, three of these showing also high CBV. Use of the present approach to determine OER for the assessment of hemodynamic adaptations in CS patients is discussed in the light of documented hemodynamic adaptations to carotid stenosis. PMID- 15110017 TI - Random field-union intersection tests for EEG/MEG imaging. AB - Electrophysiological (EEG/MEG) imaging challenges statistics by providing two views of the same spatiotemporal data: topographic and tomographic. Until now, statistical tests for these two situations have developed separately. This work introduces statistical tests for assessing simultaneously the significance of spatiotemporal event-related potential/event-related field (ERP/ERF) components and that of their sources. The test for detecting a component at a given time instant is provided by a Hotelling's T(2) statistic. This statistic is constructed in such a manner to be invariant to any choice of reference and is based upon a generalized version of the average reference transform of the data. As a consequence, the proposed test is a generalization of the well-known Global Field Power statistic. Consideration of tests at all time instants leads to a multiple comparison problem addressed by the use of Random Field Theory (RFT). The Union-Intersection (UI) principle is the basis for testing hypotheses about the topographic and tomographic distributions of such ERP/ERF components. The performance of the method is illustrated with actual EEG recordings obtained from a visual experiment of pattern reversal stimuli. PMID- 15110018 TI - Viewing the motion of human body parts activates different regions of premotor, temporal, and parietal cortex. AB - Activation of premotor and temporoparietal cortex occurs when we observe others movements, particularly relating to objects. Viewing the motion of different body parts without the context of an object has not been systematically evaluated. During a 3T fMRI study, 12 healthy subjects viewed human face, hand, and leg motion, which was not directed at or did not involve an object. Activation was identified relative to static images of the same human face, hand, and leg in both individual subject and group average data. Four clear activation foci emerged: (1) right MT/V5 activated to all forms of viewed motion; (2) right STS activated to face and leg motion; (3) ventral premotor cortex activated to face, hand, and leg motion in the right hemisphere and to leg motion in the left hemisphere; and (4) anterior intraparietal cortex (aIP) was active bilaterally to viewing hand motion and in the right hemisphere leg motion. In addition, in the group data, a somatotopic activation pattern for viewing face, hand, and leg motion occurred in right ventral premotor cortex. Activation patterns in STS and aIP were more complex--typically activation foci to viewing two types of human motion showed some overlap. Activation in individual subjects was similar; however, activation to hand motion also occurred in the STS with a variable location across subjects--explaining the lack of a clear activation focus in the group data. The data indicate that there are selective responses to viewing motion of different body parts in the human brain that are independent of object or tool use. PMID- 15110019 TI - Left hemisphere specialization for the control of voluntary movement rate. AB - Although persuasive behavioral evidence demonstrates the superior dexterity of the right hand in most people under a variety of conditions, little is known about the neural mechanisms responsible for this phenomenon. As this lateralized superiority is most evident during the performance of repetitive, speeded movement, we used parametric rate variations to compare visually paced movement of the right and left hands. Twelve strongly right-handed subjects participated in a functional magnetic resonance imaging (fMRI) experiment involving variable rate thumb movements. For movements of the right hand, contralateral rate-related activity changes were identified in the precentral gyrus, thalamus, and posterior putamen. For left-hand movements, activity was seen only in the contralateral precentral gyrus, consistent with the existence of a rate-sensitive motor control subsystem involving the left, but not the right, medial premotor corticostriatal loop in right-handed individuals. We hypothesize that the right hemisphere system is less skilled at controlling variable-rate movements and becomes maximally engaged at a lower movement rate without further modulation. These findings demonstrate that right- and left-hand movements engage different neural systems to control movement, even during a relatively simple thumb flexion task. Specialization of the left hemisphere corticostriatal system for dexterity is reflected in asymmetric mechanisms for movement rate control. PMID- 15110020 TI - The encoding of saccadic eye movements within human posterior parietal cortex. AB - Over the last few years, several functionally distinct subregions of the posterior parietal cortex (PPC) have been shown to subserve oculomotor control. Since these areas seem to overlap with regions whose activation is related to attention, we used functional magnetic resonance imaging to compare the cerebral activation pattern evoked by eye movements with different attentional loads, i.e., oscillatory saccades with different frequencies, as well as predictable, and unpredictable saccades. Our results show activation in largely overlapping networks with differing strength of activity and symmetry of involved areas. Predictable saccades having the shortest saccadic latency led to the most pronounced cerebral activity both in terms of cortical areas involved and signal intensity. Predictable and unpredictable saccades were dominated by activation within the right hemisphere, whereas oscillatory saccades showing the longest saccadic latency were dominated by activation within the left hemisphere. In all tasks, the centers of gravity of activation occurred within the posterior part of the intraparietal sulcus (IPS), while the predictable saccades additionally activated its anterior part. The enhanced activity during the execution of predictable saccades was probably related to top-down processing and/or the preparation of the upcoming eye movement. The hemispheric difference could arise from a predominant role of the right PPC for shifting spatial attention and the left PPC for shifting temporal attention. The differential encoding of saccadic eye movements within IPS indicates that the PPC splits up into different functional modules related to the particular demands of a saccade. PMID- 15110021 TI - Attentional effects of noradrenaline vary with arousal level: selective activation of thalamic pulvinar in humans. AB - Subjects sedated by noradrenergic alpha2 agonists can switch rapidly from a state of extremely low to almost full consciousness following phasic increases in arousal or cognitive demand. Such flexibility is not displayed by traditional sedatives, such as the benzodiazepine diazepam. Experimentally, the phasic modulation of alpha2 effect by arousing or distracting stimuli can counteract the deleterious cognitive effects of alpha2 agonists. We used behavioural and fMRI indices of brain function to investigate the phasic modulatory effect that presentation of loud white noise would have on attentional dysfunction induced by administration of dexmedotomidine, an alpha2 agonist. Dexmedotomidine and midazolam were compared to placebo during performance of a target detection task, which was presented in the presence or absence of white noise. Compared to placebo, both dexmedotomidine and midazolam impaired task performance. This impairment was significantly attenuated by presentation of white noise in the dexmedotomidine condition only. This functional improvement corresponded to selective increase in activity of left medial pulvinar nucleus of the thalamus. This regional increase is suggested to index increases in phasic arousal, which counteract dexmedotomidine's detrimental attentional effects. Finally, despite sedating subjects to equivalent degrees, dexmedotomidine and midazolam had strikingly different regional effects on task-induced brain activity. Therefore, for the same level of sedation, the behavioural and anatomical attributes identifying the quality of sedation can vary. PMID- 15110022 TI - Geometrical interpretation of fMRI-guided MEG/EEG inverse estimates. AB - Magneto- and electroencephalography (MEG/EEG) and functional magnetic resonance imaging (fMRI) provide complementary information about the functional organization of the human brain. An important advantage of MEG/EEG is the millisecond time resolution in detecting electrical activity in the cerebral cortex. The interpretation of MEG/EEG signals, however, is limited by the difficulty of determining the spatial distribution of the neural activity. Functional MRI can help in the MEG/EEG source analysis by suggesting likely locations of activity. We present a geometric interpretation of fMRI-guided inverse solutions in which the MEG/EEG source estimate minimizes a distance to a subspace defined by the fMRI data. In this subspace regularization (SSR) approach, the fMRI bias does not assume preferred amplitudes for MEG/EEG sources, only locations. Characteristic dependence of the source estimates on the regularization parameters is illustrated with simulations. When the fMRI locations match the true MEG/EEG source locations, they serve to bias the underdetermined MEG/EEG inverse solution toward the fMRI loci. Importantly, when the fMRI loci do not match the true MEG/EEG loci, the solution is insensitive to those fMRI loci. PMID- 15110023 TI - The effect of carbamazepine on human corticomuscular coherence. AB - EEG recordings from motor cortex show oscillations at approximately 10 and 20 Hz. The 20-Hz oscillations are coherent with contralateral EMG; in most studies those at 10 Hz are not. However, significant 10-Hz coherence has recently been reported in a group of epileptic patients, all of whom were taking the anticonvulsant drug carbamazepine (CBZ). In a double blind study, we investigated the effects of CBZ on corticomuscular coherence in eight healthy human subjects (all male). Subjects performed a precision grip task against an auxotonic load, whilst left sensorimotor EEG and EMGs from five muscles in the right hand and forearm were recorded. CBZ (100 mg) or a placebo was then given orally, and 6 h later subjects were re-tested. One week separated CBZ and placebo experiments in each subject. Coherence averaged across subjects and muscles during the hold phase of the task was maximal at 21 Hz; it increased significantly (P < 0.05, Z-test) by 89% after CBZ administration. This was significantly greater than a much smaller increase following placebo, which itself may reflect an effect of the time of day when experiments were performed. There was no significant approximately 10-Hz coherence either before or after CBZ administration. CBZ did not significantly alter EEG power at either 10 or 20 Hz. Recently, we showed that diazepam markedly increases the power of approximately 20-Hz motor cortical oscillations with little effect on coherence. We show here that CBZ raises coherence without altering EEG power. This pharmacological dissociation may indicate an important role for corticomuscular coherence in motor control. PMID- 15110024 TI - Shortening intertrial intervals in event-related cognitive studies with near infrared spectroscopy. AB - Functional near-infrared spectroscopy (fNIRS) enables imaging of brain activation by measuring changes in the concentration of oxy- and deoxy-hemoglobin (Hb). Shortening the length of the intertrial interval (ITI) seems to be a precondition for further cognitive event-related fNIRS experiments because it leads to higher statistical power. Therefore, this study investigated whether the ITI may be reduced from 12, 6, 4 s to at least 2 s. Brain activation was examined with a NIRO-300 spectrometer at the lateral prefrontal cortex in 17 healthy subjects during a randomized event-related color-word matching Stroop task. In the left lateral prefrontal cortex, the concentration of deoxy-Hb decreased significantly stronger during incongruent than neutral trials for an ITI of 12, 6, and 2 s due to coping with interference. For 4 s of ITI, no hemodynamic interference effect was detected, which was paralleled by low behavioral interference. Further, we examined whether the length of the ITI influenced the mean hemodynamic response. Shortening the ITI reduced the amplitude of oxy-Hb in contrast to deoxy-Hb, which remained almost unaltered. Summarizing results, randomized event-related cognitive fNIRS studies enable short ITIs particularly if changes in deoxy-Hb are considered. PMID- 15110025 TI - Dynamic contrast-enhanced MR imaging of the stimulated pituitary gland. AB - Apoplexy due to infarction and/or hemorrhage is a frequent complication of pituitary adenoma, occurring either spontaneously or precipitated by several factors, among them pituitary function test with hypothalamic releasing hormones. The mechanism by which releasing hormones cause pituitary apoplexy is unclear. It has been proposed that increase in pituitary size and/or alterations in blood flow could be responsible. The aim of this study was to explore the effects of intravenous administration of hypothalamic releasing hormones on pituitary size and hemodynamics in healthy subjects. Gadolinium-DTPA-enhanced dynamic magnetic resonance imaging (MRI) was performed in eight healthy volunteers under basal conditions and 20 min after injection of releasing hormones. Mean upslopes of Gadolinium-DTPA enhancement curves showed good correlation between basal and stimulated conditions (R = 0.89) and were significantly steeper after stimulation (P = 0.017). In contrast, pituitary height, width and length did not differ significantly between basal and stimulated conditions. In conclusion, the pituitary does not swell in healthy subjects in response to stimulation with hypothalamic releasing hormones, whereas transfer of contrast agent to tissue (blood flow and/or vessel permeability) is enhanced. PMID- 15110026 TI - Grey and white matter atrophy in early clinical stages of primary progressive multiple sclerosis. AB - BACKGROUND: There is little information available on grey and white matter (GM and WM) atrophy in primary progressive multiple sclerosis (PPMS) and on their relationships with clinical and other magnetic resonance imaging (MRI) measures. AIM: To evaluate disease progression in the early phase of PPMS, focusing on axonal loss as assessed by volumetric MRI measures of WM and GM, and to determine their relationships with clinical outcomes and lesion load measures. METHODS: Forty-three patients with PPMS within 5 years of symptom onset and 45 control subjects were studied. Three-dimensional brain scans were acquired and segmented into WM, GM, and cerebrospinal fluid (CSF) using SPM99. Brain parenchymal (BPF), WM (WMF), and GM fractions (GMF) normalized against total intracranial volumes were estimated. T2-weighted (T2) and enhancing lesion loads were also determined. Expanded Disability Status Scale (EDSS) and Multiple Sclerosis Functional Composite (MSFC) scores were recorded in all patients. RESULTS: There were significant differences between patients and controls in BPF, WMF, and GMF values (P < 0.001). BPF (r = -0.469; P = 0.002) and WMF (r = -0.532; P < 0.001) but not GMF (r = -0.195; P = 0.2) correlated with EDSS scores. BPF (r = 0.518; P = 0.001), WMF (r = 0.483; P = 0.001), and GMF (r = 0.337; P = 0.031) correlated with MSFC scores. Correlations with enhancing lesion and T2 loads were only significant for BPF and WMF. CONCLUSIONS: Brain atrophy is seen in the early stages of PPMS and affects both GM and WM. WM atrophy appears more closely related to clinical outcome and WM focal damage than GM atrophy in this patient group. PMID- 15110027 TI - A method for removal of global effects from fMRI time series. AB - We present a technique for removing global effects from functional magnetic resonance imaging (fMRI) images, using a voxel-level linear model of the global signal (LMGS). The procedure does not assume low-frequency global effects and is based on the assumption that the global signal (the time course of the average intensity per volume) is replicated in the same pattern throughout the brain, although not necessarily at the same magnitude. A second assumption is that all effects that match the global signal are of no interest and can be removed. The method involves modeling the time course of each voxel to the global signal and removing any such global component from the voxel's time course. A challenge that elicits a large change in the global blood oxygenation level-dependent (BOLD) signal, inspired hypercapnia (5% CO(2)/95% O(2)), was administered to 14 subjects during a 144-s, 24-scan fMRI procedure; baseline series were also collected. The method was applied to these data and compared to intensity normalization and low frequency spline detrending. A large global BOLD signal increase emerged to the hypercapnic challenge. Intensity normalization failed to remove global components due to regional variability. Both LMGS and spline detrending effectively removed low-frequency components, but unlike spline detrending (which is designed to remove only low frequency trends), the LMGS removed higher-frequency global fluctuations throughout the challenge and baseline series. LMGS removes all effects correlated with the global signal, and may be especially useful for fMRI data that include large global effects and for generating detrended images to use with subsequent volume-of-interest (VOI) analyses. PMID- 15110028 TI - Improved agreement between Talairach and MNI coordinate spaces in deep brain regions. AB - Disagreement between the Talairach atlas and the stereotaxic space commonly used in software like SPM is a widely recognized problem. Others have proposed affine transformations to improve agreement in surface areas such as Brodmann's areas. This article proposes a similar transformation with the goal of improving agreement specifically in the deep brain region. The task is accomplished by finding an affine transformation that minimizes the mean distance between the surface coordinates of the lateral ventricles in the Talairach atlas and the MNI templates. The result is a transformation that improves deep brain agreement over both the untransformed Talairach coordinates and the surface-oriented transformation. While the transformation improves deep brain agreement, surface agreement is generally made worse. For areas near the lateral ventricle, the transformation presented herein is valuable for applications such as region of interest (ROI) modeling. PMID- 15110029 TI - Dissociable amygdala and orbitofrontal responses during reversal fear conditioning. AB - The neural mechanisms underlying the persistence and plasticity of human emotional learning are unknown. Here we describe dissociable neural responses in amygdala and orbitofrontal cortex during acquisition and reversal of discriminatory fear conditioning. During acquisition, increased responses in bilateral amygdala were elicited by a face stimulus (A = CS+) predictive of an aversive noise compared to another nonpredictive face (B = CS-). With subsequent reversal of the conditioning contingency, face B (new CS+) elicited enhanced responses in right orbitofrontal cortex, while face A (old CS+) continued to evoke increased responses in right ventral amygdala. Thus, while orbitofrontal cortex exhibited rapid reversal of acquired fear responses, ventral amygdala showed a persistent, nonreversing "memory" for previous fear-related stimulus associations. PMID- 15110030 TI - In vivo 2D J-resolved magnetic resonance spectroscopy of rat brain with a 3-T clinical human scanner. AB - A clinical 3-T scanner equipped with a custom-made transmit/receive birdcage coil was used to collect 2D J-resolved single-voxel spectroscopy in vivo of rat brain. Four adult Wistar rats were scanned twice each, with a 2-week interval. Voxel size was approximately 5 x 10 x 5 mm(3). Total spectroscopic acquisition time was 14 min for collection of two 4:20 min water-suppressed acquisitions and one 4:20 min acquisition acquired in the absence of water suppression. The unsuppressed water data were used in post-processing to reduce residual water side bands, as well as for metabolite signal normalization to account for variations in coil loading and voxel size. Peak areas were estimated for resonances from N-acetyl aspartate (NAA), creatine, choline, taurine, glutamate, and combined glutamate and glutamine. T(2)-relaxation times were estimated for NAA and creatine. The average deviation from the mean of repeated measures for glutamate, combined glutamate and glutamine, and taurine ranged from 7.6% to 18.3%, while for NAA, creatine, and choline, the deviation was less than 3%. The estimated T(2) values for NAA (mean +/- SD = 330 +/- 57 ms) and creatine (174 +/- 27 ms) were similar to those reported previously for rat brain and for human gray and white matter. These results indicate that reliable, small animal brain MR spectroscopy can be performed on a human clinical 3-T scanner. PMID- 15110031 TI - Short-term (approximately 600 ms) prediction of perturbation dynamics for 10- and 20-Hz MEG rhythms in human primary sensorimotor hand cortices. AB - The pericentral primary sensorimotor cortices generate the "mu rhythm" with a distinct spectral signature exhibiting two peaks, generated predominantly anterior (20 Hz) or posterior (10 Hz) to the central sulcus; it defines a "background" network state upon which somatosensory inputs will impinge. We used the high spatiotemporal resolution of magnetoencephalography to analyze the perturbation dynamics of these cortical rhythms in response to a series of paired electric median nerve stimuli: single trials were sorted off-line according to increasing power of the 10- or 20-Hz rebounds which occurred 300-600 ms after the first stimulus; using subaverages formed from the upper and lower 20% of this distribution, we analyzed somatosensory evoked fields (SEF) and power modifications caused by the second stimulus in the pair. We report three key findings: (1) the power level of rhythm rebounds triggered by the first stimulus predicted the rebound strength after the second stimulus applied 600 ms later; yet, it was uncorrelated across the 2.4-s interval separating subsequent stimulus pairs. (2) Conventional averaging camouflages substantial trial-to-trial variations of rhythm dynamics including, for example, even non-occurrences of rhythm rebounds. (3) For six of the seven subjects, the background rhythm power did not affect any SEF component; for the subject with the strongest rhythms only intracortically generated deflections (peaking after the thalamocortical input component N20m) varied as function of pre-stimulus 10- or 20-Hz power. Thus, the perturbation dynamics of the pericentral mu rhythm exhibits a significant intertrial variance, which becomes effective mainly at a time scale larger than 600 ms. PMID- 15110032 TI - Regional homogeneity approach to fMRI data analysis. AB - Kendall's coefficient concordance (KCC) can measure the similarity of a number of time series. It has been used for purifying a given cluster in functional MRI (fMRI). In the present study, a new method was developed based on the regional homogeneity (ReHo), in which KCC was used to measure the similarity of the time series of a given voxel to those of its nearest neighbors in a voxel-wise way. Six healthy subjects performed left and right finger movement tasks in event related design; five of them were additionally scanned in a rest condition. KCC was compared among the three conditions (left finger movement, right finger movement, and the rest). Results show that bilateral primary motor cortex (M1) had higher KCC in either left or right finger movement condition than in rest condition. Contrary to prediction and to activation pattern, KCC of ipsilateral M1 is significantly higher than contralateral M1 in unilateral finger movement conditions. These results support the previous electrophysiologic findings of increasing ipsilateral M1 excitation during unilateral movement. ReHo can consider as a complementary method to model-driven method, and it could help reveal the complexity of the human brain function. More work is needed to understand the neural mechanism underlying ReHo. PMID- 15110033 TI - Assessing the auditory dual-pathway model in humans. AB - Evidence from anatomical and neurophysiological studies in nonhuman primates suggests a dual-pathway model of auditory processing wherein sound identity and sound location information are segregated along ventral and dorsal streams, respectively. The present meta-analysis reviewed evidence from auditory functional magnetic resonance imaging (fMRI) and positron emission tomography (PET) studies to determine the reliability of this model in humans. Activation coordinates from 11 "spatial" studies (i.e., listeners made localization judgements on sounds that could occur at two or more perceptually different positions) and 27 "nonspatial" studies (i.e., listeners completed nonspatial tasks involving sounds presented from the same location) were entered into the analysis. All but one of the spatial studies reported activation within the inferior parietal lobule as opposed to only 41% of the nonspatial studies. In addition, 55% of spatial studies reported activity around the superior frontal sulcus as opposed to only 7% of the nonspatial studies. In comparison, inferior frontal activity (Brodmann's areas 45 and 47) was reported in only 9% of the spatial studies, but in 56% of the nonspatial studies. Finally, almost all temporal lobe activity observed during spatial tasks was confined to posterior areas, whereas nonspatial activity was distributed throughout the temporal lobe. These results support an auditory dual-pathway model in humans in which nonspatial sound information (e.g., sound identity) is processed primarily along the ventral stream whereas sound location is processed along the dorsal stream and areas posterior to primary auditory cortex. PMID- 15110034 TI - Limbic-frontal circuitry in major depression: a path modeling metanalysis. AB - This paper reports the results of an across lab metanalysis of effective connectivity in major depression (MDD). Using FDG PET data and Structural Equation Modeling, a formal depression model was created to explicitly test current theories of limbic-cortical dysfunction in MDD and to characterize at the path level potential sources of baseline variability reported in this patient population. A 7-region model consisting of lateral prefrontal cortex (latF9), anterior thalamus (aTh), anterior cingulate (Cg24), subgenual cingulate (Cg25), orbital frontal cortex (OF11), hippocampus (Hc), and medial frontal cortex (mF10) was tested in scans of 119 depressed patients and 42 healthy control subjects acquired during three separate studies at two different institutions. A single model, based on previous theory and supported by anatomical connectivity literature, was stable for the three groups of depressed patients. Within the context of this model, path differences among groups as a function of treatment response characteristics were also identified. First, limbic-cortical connections (latF9-Cg25-OF11-Hc) differentiated drug treatment responders from nonresponders. Second, nonresponders showed additional abnormalities in limbic-subcortical pathways (aTh-Cg24-Cg25-OF11-Hc). Lastly, more limited limbic-cortical (Hc-latF9) and cortical-cortical (OF11-mF10) path differences differentiated responders to cognitive behavioral therapy (CBT) from responders to pharmacotherapy. We conclude that the creation of such models is a first step toward full characterization of the depression phenotype at the neural systems level, with implications for the future development of brain-based algorithms to determine optimal treatment selection for individual patients. PMID- 15110035 TI - The chronoarchitecture of the human brain--natural viewing conditions reveal a time-based anatomy of the brain. AB - A dominant tendency in cerebral studies has been the attempt to locate architecturally distinct parts of the cortex and assign special functions to each, through histological, clinical or hypothesis-based imaging experiments. Here we show that the cerebral cortex can also be subdivided into different components temporally, without any a priori hypotheses, based on the principle of functional independence. This states that distinct functional subdivisions have activity time courses (ATCs) that are, if not independent, at least characteristic to each when the brain is exposed to natural conditions. To approach a time-based anatomy experimentally, we recorded whole-brain activity using functional magnetic resonance imaging (fMRI) and analyzed the data with independent component analysis (ICA). Our results show that a multitude of cortical areas can be identified based purely on their characteristic ATCs during natural conditions. We demonstrate that a more "rich" stimulation (free viewing of a movie) leads to more areas being activated in a specific way than conventional stimuli, allowing for a more detailed dissection of the cortex into its subdivisions. We show that stimulus-driven functionally specialized areas can be identified by intersubject correlation even if their function is unknown. Chronoarchitectonic mapping thus opens the prospect of identifying previously unknown cortical subdivisions based on natural viewing conditions by exploiting the characteristic temporal "fingerprint" that is unique to each. PMID- 15110036 TI - Fast and robust registration of PET and MR images of human brain. AB - In recent years, mutual information has proved to be an excellent criterion for registration of intra-individual images from different modalities. Multi resolution coarse-to-fine optimization was proposed for speeding-up of the registration process. The aim of our work was to further improve registration speed without compromising robustness or accuracy. We present and evaluate two procedures for co-registration of positron emission tomography (PET) and magnetic resonance (MR) images of human brain that combine a multi-resolution approach with an automatic segmentation of input image volumes into areas of interest and background. We show that an acceleration factor of 10 can be achieved for clinical data and that a suitable preprocessing can improve robustness of registration. Emphasis was laid on creation of an automatic registration system that could be used routinely in a clinical environment. For this purpose, an easy to-use graphical user interface has been developed. It allows physicians with no special knowledge of the registration algorithm to perform a fast and reliable alignment of images. Registration progress is presented on the fly on a fusion of images and enables visual checking during a registration. PMID- 15110037 TI - CBF changes during brain activation: fMRI vs. PET. AB - The changes in regional cerebral blood flow (rCBF) associated with the changes in neuronal activity are routinely measured both by positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) techniques. However, direct comparison has not been performed to determine similarities and differences of PET and fMRI techniques in determining the rCBF response to brain activation. In the present study, a quantitative comparison of the functional rCBF maps obtained by PET and fMRI are made by performing an activation study in a single group of subjects under precisely controlled conditions and using identical visual stimuli. Twelve healthy volunteers participated in the activation study using the visual checkerboard stimulation with flip frequency at 8 Hz. By selecting the conjunctive pixels which activated on both PET and fMRI maps, the change in rCBF measured by fMRI was 36.95 +/- 2.54%, whereas the value measured by PET was 38.79 +/- 2.63%. Our results have demonstrated that there is no statistically significant difference (P = 0.22) in the measurements of rCBF change between MRI and PET methods. PMID- 15110038 TI - The neural correlates of placebo effects: a disruption account. AB - The neurocognitive pathways by which placebo effects operate are poorly understood. Positron emission tomography (PET) imaging was used to assess the brain response of patients with chronic abdominal pain (irritable bowel syndrome; IBS) to induced intestinal discomfort both before and after a 3-week placebo regimen. A daily symptom diary was used to measure symptom improvement. Increases in right ventrolateral prefrontal cortex (RVLPFC) activity from pre- to post placebo predicted self-reported symptom improvement, and this relationship was mediated by changes in dorsal anterior cingulate (dACC), typically associated with pain unpleasantness. These results are consistent with disruption theory [Lieberman, M.D., 2003. Reflective and reflexive judgment processes: a social cognitive neuroscience approach. In: Forgas, J.P., Williams, K.R., von Hippel, W. (Eds.), Social Judgments: Explicit and Implicit Processes. Cambridge Univ. Press, New York, pp. 44-67], which proposes that activation of prefrontal regions associated with thinking about negative affect can diminish dACC and amygdala reactivity to negative affect stimuli. This is the first study to identify a neural pathway from a region of the brain associated with placebos and affective thought to a region closely linked to the placebo-related outcome of diminished pain unpleasantness. PMID- 15110039 TI - Olfactory system activation from sniffing: effects in piriform and orbitofrontal cortex. AB - Neuroimaging studies suggest that piriform cortex is activated at least in part by sniffing. We used H(2)(15)O positron emission tomography (PET) to study 15 healthy volunteers while they participated in four conditions, two of which were sniffing odorants and odorless air. The remaining two conditions involved a constant, very low flow of either odorized or odorless air during velopharyngeal closure (VPC), a technique that prevents subject-induced airflow through the nasal passages. Contrary to expectation, sniffing under odorless conditions did not induce significant piriform and surrounding cortical (PC+) activity when compared to odorless VPC, even at a liberal statistical threshold. However, a small correlation emerged in PC+ between the difference signal of [odorless sniffing - odorless VPC] and peak rate of nasal pressure change. PC+ activity was, however, strongly evoked by odorant exposure during sniffing and VPC, with neither technique showing greater activation. Activity in orbitofrontal (olfactory association) cortex was absent during odorant stimulation (OS) with VPC, but present during odorant sniffing. Sniffing may therefore play an important role in facilitating the higher-order analysis of odors. A right orbitofrontal region was also activated with odorless sniffing, which suggests a possible orbitofrontal role in guided olfactory exploration. PMID- 15110040 TI - Specialization within the ventral stream: the case for the visual word form area. AB - Is there specialization for visual word recognition within the visual ventral stream of literate human adults? We review the evidence for a specialized "visual word form area" and critically examine some of the arguments recently placed against this hypothesis. Three distinct forms of specialization must be distinguished: functional specialization, reproducible localization, and regional selectivity. Examination of the literature with this theoretical division in mind indicates that reading activates a precise subpart of the left ventral occipitotemporal sulcus, and that patients with pure alexia consistently exhibit lesions of this region (reproducible localization). Second, this region implements processes adequate for reading in a specific script, such as invariance across upper- and lower-case letters, and its lesion results in the selective loss of reading-specific processes (functional specialization). Third, the issue of regional selectivity, namely, the existence of putative cortical patches dedicated to letter and word recognition, cannot be resolved by positron emission tomography or lesion data, but requires high-resolution neuroimaging techniques. The available evidence from single-subject fMRI and intracranial recordings suggests that some cortical sites respond preferentially to letter strings than to other categories of visual stimuli such as faces or objects, though the preference is often relative rather than absolute. We conclude that learning to read results in the progressive development of an inferotemporal region increasingly responsive to visual words, which is aptly named the visual word form area (VWFA). PMID- 15110041 TI - The pro and cons of labelling a left occipitotemporal region: "the visual word form area". PMID- 15110042 TI - Involvement of Islet-2 in the Slit signaling for axonal branching and defasciculation of the sensory neurons in embryonic zebrafish. AB - In Drosophila melanogaster, Slit acts as a repulsive cue for the growth cones of the commissural axons which express a receptor for Slit, Roundabout (Robo), thus preventing the commissural axons from crossing the midline multiple times. Experiments using explant culture have shown that vertebrate Slit homologues also act repulsively for growth cone navigation and neural migration, and promote branching and elongation of sensory axons. Here, we demonstrate that overexpression of Slit2 in vivo in transgenic zebrafish embryos severely affected the behavior of the commissural reticulospinal neurons (Mauthner neurons), promoted branching of the peripheral axons of the trigeminal sensory ganglion neurons, and induced defasciculation of the medial longitudinal fascicles. In addition, Slit2 overexpression caused defasciculation and deflection of the central axons of the trigeminal sensory ganglion neurons from the hindbrain entry point. The central projection was restored by either functional repression or mutation of Robo2, supporting its role as a receptor mediating the Slit signaling in vertebrate neurons. Furthermore, we demonstrated that Islet-2, a LIM/homeodomain-type transcription factor, is essential for Slit2 to induce axonal branching of the trigeminal sensory ganglion neurons, suggesting that factors functioning downstream of Islet-2 are essential for mediating the Slit signaling for promotion of axonal branching. PMID- 15110043 TI - The 5-HT receptor cell is a new member of secondary mesenchyme cell descendants and forms a major blastocoelar network in sea urchin larvae. AB - A gene encoding the serotonin (5-hydroxytryptamine, 5-HT) receptor (5-HT-hpr) was identified from the sea urchin, Hemicentrotus pulcherrimus. Partial amino acid sequence deduced from the cDNA showed strong similarity to Aplysia californica 5 HT2 receptor. Immunoblotting analysis of this 5-HT-hpr protein (5-HT-hpr) with an antibody raised against a deduced peptide showed two bands. Their relative molecular masses were 69 and 53 kDa, respectively. The larger band alone disappeared after N-glycopeptidase F digestion, indicating the protein was N glycosylated. Immunolocalization analysis showed that cells expressing the 5-HT hpr (SRC) first appeared near the tip of the archenteron in 33-h post fertilization (33 hpf) prism larvae. Their cell number doubled in 2 h, and 5-HT hpr protein expression increased further without cell proliferation. SRC spread ventrally on the basal surface of the oral ectoderm in 36 hpf prism larvae, and then clockwise on the ventral ectoderm to the posterior region to complete formation of the SRC network in 48 hpf early plutei. The SRC network was comprised of 7 main tracts: 4 spicule system-associated tracts and 3 spicule system-independent tracts. The network extended short fibers to the larval body surface through the ectoderm, implicating a signal transmission system that receives exogenous signal. Double-stain immunohistochemistry with antibodies to primary mesenchyme cells showed that SRC were not stained by the antibody. In embryos deprived of secondary mesenchyme cell (SMC) by microsurgery, the number of SRC decreased considerably. These two data indicate that SRC are SMC descendants, adding a new member to the SMC lineage. PMID- 15110044 TI - Feedback mechanisms regulate retinoic acid production and degradation in the zebrafish embryo. AB - Retinoic acid (RA) signaling in vertebrate embryos occurs in a distinct physical and temporal pattern. Regulating this spatial distribution is crucial to the development of the embryo, as RA in excess or in inappropriate tissues is teratogenic. In order to understand how RA availability is determined in zebrafish we have investigated the expression of cyp26a1, an enzyme that inactivates RA, and its relationship to raldh2, one of the enzymes that produce RA from retinal. cyp26a1 expression follows three phases: in presumptive anterior neurectoderm and in a circumblastoporal ring during gastrulation, in the tailbud throughout somitogenesis, and in multiple specific tissue types beginning at mid somitogenesis and continuing through 48 h postfertilization (hpf). This expression was either adjacent or opposite to those tissues expressing raldh2. We then investigated how RA production might regulate these relationships. Endogenous RA produced by raldhs did not play a role in setting cyp26a1 expression in most tissues. However, exogenous RA regulates expression of both enzymes. cyp26a1 is up regulated in the embryo in a time, concentration, and tissue-dependent manner. Conversely, raldh2 expression is reduced with RA treatment. Tests of the raldh2 promoter in cell transfections proved that RA directly represses its activity. These data demonstrate that the feedback mechanisms regulating production and degradation of RA must be considered in any experiments altering levels of RA in the developing vertebrate embryo. PMID- 15110045 TI - Drosophila genetic variants that change cell size and rate of proliferation affect cell communication and hence patterning. AB - We explore in this paper the role of genetic variants that affect cell size and proliferation in the determination of organ size. We use genetic mosaics of loss or gain of function in six different loci, which promotes smaller or larger than normal cells, associated to either smaller or larger than normal territories. These variants have autonomous effects on patterning and growth in mutant territories. However, there is no correlation between cell size or rate of proliferation on the size of the mutant territory. In addition, these mosaics show non-autonomous effects on surrounding wildtype cells, consisting always in a reduction in number of non-mutant cells. In all mutant conditions the final size (and shape) of the wing is different than normal. The phenotypes of the same variants include higher density of chaetae in the notum. These autonomous and non autonomous effects suggest that the control of size in the wing is the result of local cell communication defining canonic distances between cells in a positional values landscape. PMID- 15110046 TI - Dorso-ventral asymmetric functions of teashirt in Drosophila eye development depend on spatial cues provided by early DV patterning genes. AB - The teashirt (tsh) gene has dorso-ventral (DV) asymmetric functions in Drosophila eye development: promoting eye development in dorsal and suppressing eye development in ventral by Wingless mediated Homothorax (HTH) induction [Development 129 (2002) 4271]. We looked for DV spatial cues required by tsh for its asymmetric functions. The dorsal Iroquois-Complex (Iro-C) genes and Delta (Dl) are required and sufficient for the tsh dorsal functions. The ventral Serrate (Ser), but not fringe (fng) or Lobe (L), is required and sufficient for the tsh ventral function. We propose that DV asymmetric function of tsh represents a novel tier of DV pattern regulation, which takes place after the spatial expression patterns of early DV patterning genes are established in the eye. PMID- 15110047 TI - Genetic evidence for involvement of maternally derived Wnt canonical signaling in dorsal determination in zebrafish. AB - In zebrafish, the program for dorsal specification begins soon after fertilization. Dorsal determinants are localized initially to the vegetal pole, then transported to the blastoderm, where they are thought to activate the canonical Wnt pathway, which induces the expression of dorsal-specific genes. We identified a novel maternal-effect recessive mutation, tokkaebi (tkk), that affects formation of the dorsal axis. Severely ventralized phenotypes, including a lack of dorso-anterior structures, were seen in 5-100% of the embryos obtained from tkk homozygous transmitting females. tkk embryos displayed defects in the nuclear accumulation of beta-catenin on the dorsal side, and reduced or absent expression of dorsal-specific genes. Mesoderm and endoderm formation outside the dorsal axis was not significantly affected. Injection of RNAs for activated beta catenin, dominant-negative forms of Axin1 and GSK3beta, and wild-type Dvl3, into the tkk embryos suppressed the ventralized phenotypes and/or dorsalized the embryos, and restored or induced an ectopic and expanded expression of bozozok/dharma and goosecoid. However, dorsalization by wnt RNAs was affected in the tkk embryos. Inhibition of cytoplasmic calcium release elicited an ectopic and expanded expression of chordin in the wild-type, but did not restore chordin expression efficiently in the tkk embryos. These data indicate that the tkk gene product functions upstream of or parallel to the beta-catenin-degradation machinery to control the stability of beta-catenin. The tkk locus was mapped to chromosome 16. These data provide genetic evidence that the maternally derived canonical Wnt pathway upstream of beta-catenin is involved in dorsal axis formation in zebrafish. PMID- 15110048 TI - Endothelin-1 regulates the dorsoventral branchial arch patterning in mice. AB - Endothelin-1 (ET-1), a 21-amino acid peptide secreted by the epithelium and core mesenchyme in the branchial arches as well as vascular endothelium, is involved in craniofacial and cardiovascular development through endothelin receptor type-A (EdnrA) expressed in the neural crest-derived ectomesenchyme. Here we show that ET-1(-/-) mutant mice exhibit a homeotic-like transformation of the lower jaw to an upper jaw. Most of the maxillary arch-derived components are duplicated and replaced mandibular arch-derived structures, resulting in a mirror image of the upper and lower jaws in the ET-1(-/-) mutant. As for hyoid arch-derivatives, the ventral structures are severely affected in comparison to the dorsal ones in the ET-1(-/-) mutant. Correspondingly, the expression of Dlx5 and Dlx6, Distalless related homeobox genes determining the ventral identity of the anterior branchial arches, and of the mandibular marker gene Pitx1 is significantly downregulated in the ET-1(-/-) mutant, whereas the expression of Dlx2 and the maxillary marker gene Prx2 is unaffected or rather upregulated. These findings indicate that the ET-1/EdnrA signaling may contribute to the dorsoventral axis patterning of the branchial arch system as a mediator of the regional intercellular interactions. PMID- 15110049 TI - Constitutively active PTH/PTHrP receptor in odontoblasts alters odontoblast and ameloblast function and maturation. AB - Parathyroid hormone (PTH)-related protein (PTH-rP) is an important autocrine/paracrine attenuator of programmed cell differentiation whose expression is restricted to the epithelial layer in tooth development. The PTH/PTHrP receptor (PPR) mRNA in contrast is detected in the dental papilla, suggesting that PTHrP and the PPR may modulate epithelial-mesenchymal interactions. To explore the possible interactions, we studied the previously described transgenic mice in which a constitutively active PPR is targeted to osteoblastic cells. These transgenic mice have a vivid postnatal bone and tooth phenotype, with normal tooth eruption but abnormal, widened crowns. Transgene mRNA expression was first detected at birth in the dental papilla and, at 1 week postnatally, in odontoblasts. There was no transgene expression in ameloblasts or in other epithelial structures. Prenatally, transgenic molars and incisors revealed no remarkable change. By the age of 1 week, the dental papilla was widened, with disorganization of the odontoblastic layer and decreased dentin matrix. In addition, the number of cusps was abnormally increased, the ameloblastic layer disorganized, and enamel matrix decreased. Odontoblastic and, surprisingly, ameloblastic cytodifferentiation was impaired, as shown by in situ hybridization and electron microscopy. Interestingly, ameloblastic expression of Sonic Hedgehog, a major determinant of ameloblastic cytodifferentiation, was dramatically altered in the transgenic molars. These data suggest that odontoblastic activation of the PPR may play an important role in terminal odontoblastic and, indirectly, ameloblastic cytodifferentiation, and describe a useful model to study how this novel action of the PPR may modulate mesenchymal/epithelial interactions at later stages of tooth morphogenesis and development. PMID- 15110050 TI - Health promotion in physician organizations: results from a national study. AB - BACKGROUND: Health promotion programs can be effective in improving the delivery of clinical preventive services and in improving population health; however, the availability of health promotion programs offered through physician organizations, such as medical groups and independent practice associations, are largely unknown. METHODS: This research uses data from the National Study of Physician Organizations and the Management of Chronic Illness, conducted by the University of California, Berkeley, to document the extent to which physician organizations offer health promotion programs. Of 1587 physician organizations nationally with 20 or more physicians, 1104 participated, for a response rate of 70%. RESULTS: Overall, 60% of physician organizations offer at least one health promotion program targeting one or more of eight areas: prenatal education (42%), smoking cessation (39%), nutrition (39%), weight loss (34%), health risk assessments (25%), stress management (25%), substance abuse (20%), and sexually transmitted disease prevention (16%). Factors positively associated with offering health promotion programs include the following: outside reporting of quality measures, public recognition for quality measures, clinical information technology systems, being a medical group, and ownership by a hospital or health plan. CONCLUSIONS: Physician organizations in the United States have a long way to go in offering these important programs to their patients. However, our findings also suggest that health plans, purchasers, and policymakers can play a positive role in increasing the use of these programs. By offering recognition and incentives for quality improvement, and by funding the expansion of information technology, the healthcare community can encourage and enable physician organizations to increase the availability of health promotion programs nationally. PMID- 15110051 TI - Operational conditions affecting the vaccination of older adults. AB - BACKGROUND: The content and context of the process of vaccinating older adults against influenza in outpatient settings has not been adequately described. Failure to appreciate the causal antecedents or precursors to the act of provider recommendation may explain why so many efficacious interventions identified by the U.S. Task Force on Community Preventive Services fail to be routinely implemented and why influenza immunization rates have remained static over the past decade. METHODS: This study used critical path analysis from data collected during standardized workflow observations of patients more than 50 years of age from a convenience sample of 16 ambulatory care settings in San Diego, California; Rochester, New York; and Albuquerque, New Mexico. Observations were made from October 23, 2001 to January 31, 2002. RESULTS: In this study, 62% (151/243) of patients observed during scheduled extended visits received influenza vaccinations. When operational, temporal, and clinical factors are examined altogether through critical path analysis, a model of seven critical organizational support, temporal, and clinical activities emerges that is able to predict 93% of the immunizations. Variation from the model predicts 73% of the missed opportunities. CONCLUSIONS: Vaccination of adults should not be seen as simply an incremental activity added to the general health encounter. Assuring a high rate of vaccination requires adequate time and operational support. Provider patient discussion is more productively viewed as the culmination of the immunization process, not the beginning. Finally, this study indicates the potential need to identify and compare processes of care associated with other specific preventive services. PMID- 15110052 TI - Community-based screening for cardiovascular disease and diabetes using HbA1c. AB - BACKGROUND: Persons from inner-city immigrant and mixed-ethnic communities are known to be at high risk for cardiovascular disease and diabetes. Such communities may also be underserved for preventive medical care. The authors hypothesized that hemoglobin A1c (HbA1c) could be used as a screening test for a community-based program to detect new cases of diabetes and persons at risk for diabetes and cardiovascular disease. METHODS: Screenings took place in churches, group homes, shelters, community centers, and street corners of the Bronx. Screening data included history of diabetes, age, ethnicity, body mass index, blood pressure, lipid panel, random glucose, and HbA1c. Data were analyzed for number of cases of new diabetes (HbA1c > or =7%), for patients at risk for diabetes (HbA1c 6%-6.99%), and for associations between HbA1c and other variables. The effect of location of screening and self-reported ethnicity on outcome variables was also analyzed. RESULTS: Seven hundred four persons were screened in 25 different sessions. HbA1c and lipid profile were obtained on 539 persons, which formed the cohort for analysis. Mean HbA1c for the cohort was 6.00%. Thirty-two percent of the cohort had HbA1c of more than 6%, and 11.4% had HbA1c of more than 7%. Excluding known diabetics (13% of cohort), 24% had HbA1c of more than 6%, and 3.4% had HbA1c of more than 7%. HbA1c was significantly correlated with total cholesterol, triglycerides, low-density lipoprotein, systolic blood pressure, body mass index, and age; in all cases, correlation coefficients were higher with HbA1c than with random glucose. In addition, significantly higher cardiovascular disease risk factors were found in persons with HbA1c of more than 6%; 6% may be a threshold value for the metabolic syndrome. Mean HbA1c was higher in persons from the South Bronx (which has a higher poverty rate) than the North Bronx (6.08% v 5.74%, p=0.013). There were no statistically significant differences between self-reported ethnic groupings. CONCLUSIONS: There was a high prevalence of undiagnosed diabetes, and of patients at risk for diabetes, in this community setting. Community-based screening can be used as a method for identifying high percentages of patients at risk for diabetes or with undiagnosed diabetes in an inner city, immigrant, mixed-ethnic population. PMID- 15110053 TI - Compliance to a Workplace Violence Prevention Program in small businesses. AB - BACKGROUND: Robberies are the leading motive for work-related homicide and assault. Interventions to reduce robberies and related injuries have been limited to convenience stores, and evaluations have not addressed compliance as a factor in program effectiveness. SETTING/PARTICIPANTS: A total of 314 intervention and 96 control businesses were included in this intervention evaluation. INTERVENTION: The Workplace Violence Prevention Program provided a customized robbery and violence prevention program to a stratified random sample of 314 small, high-risk businesses in Los Angeles City. An additional 96 comparison businesses did not receive the intervention. The intervention included individualized consultation, printed materials, training brochures, and a video. Interventions were conducted from August 1997 through August 2000. MAIN OUTCOME MEASURES: For both intervention and comparison businesses, a comprehensive security program assessment was conducted at baseline and at 3- and 12-month follow-up visits. Crime rates in intervention and comparison businesses were examined for 12 months pre- and post-intervention with the use of police reports. RESULTS: By the second follow-up visit, compliance to the intervention program was significant for each program component. Employee training was the most frequently implemented intervention component. Neighborhood crime level, primary language spoken by the business owner, and the number of employees were all related to compliance. Although crime rates generally increased for all businesses from the pre- to post-intervention periods, businesses with high compliance to the program experienced a decrease in overall violent crime and robbery. CONCLUSIONS: Participating businesses were willing to voluntarily implement components of the intervention program, and greater implementation was related to reductions in robbery and violent crime. PMID- 15110054 TI - Mental health impact of 9/11 Pentagon attack: validation of a rapid assessment tool. AB - BACKGROUND: Following the tragic events of 9/11/2001, the Pentagon Post Disaster Health Assessment (PPDHA) survey was created to identify healthcare needs and concerns among Pentagon personnel and to assure that appropriate care and information was provided. The PPDHA was fielded from October 15, 2001, to January 15, 2002. Fundamental in this assessment was the evaluation of the mental health impact as a result of the attack. METHODS: Although a number of standardized instruments exist for mental health domains, most are lengthy and could not be used as a rapid health assessment. Instead, a short screening instrument consisting of 17 questions was developed that covered important mental health symptom domains, mental health functioning, and possible predictive risk factors. High-risk groups for post-traumatic stress disorder (PTSD), depression, panic attacks, generalized anxiety, and alcohol abuse were assessed, and validation of risk groups was assessed across functional levels. RESULTS: Overall, 1837 (40%) respondents met the screening criteria for any of the symptom domains of interest 1 to 4 months after the attack: PTSD (7.9%), depression (17.7%), panic attacks (23.1%), generalized anxiety (26.9%), or alcohol abuse (2.5%). Mental health risk groups were highly correlated with self-reported reduced daily functioning and use of counseling services. Additionally, risk factors known to be associated with mental health problems after traumatic events were strongly predictive of the high-risk categories identified. CONCLUSIONS: Mental health concerns were common among Pentagon employees in the 4 months after the 9/11 attack. Data from this study suggested that the short mental health screening instrument had validity and can serve as a prototype for rapid public health assessment of the mental health impact of future traumatic events. PMID- 15110055 TI - Trends in alcohol use and binge drinking, 1985-1999: results of a multi-state survey. AB - BACKGROUND: Alcohol abuse is a major public health problem in the United States. Binge drinking and drinking among youth are of special concern. The purpose of this study is to examine trends in alcohol use and binge drinking and correlates of the behaviors with a focus on drinking among persons 18 to 20 years of age. METHODS: Data are from telephone interviews of 449,110 adults aged > or =18 years residing in the 19 states that participated in the Behavioral Risk Factor Surveillance System (BRFSS) from 1985 to 1999. The percentages reporting current alcohol use and binge use (> or =5 drinks per occasion) were calculated by year, age, gender, race, and level of education. Data were analyzed in 2003. RESULTS: From 1985 to 1999, the prevalence of current alcohol use dropped 7.3%, and binge drinking dropped 3.3%. Among all age groups, most of the decline occurred before 1990. The greatest decline in both current (12.6%) and binge use (7.3%) occurred in the 18- to 20-year-old group. Between 1997 and 1999, however, respondents in this age group reported increases in these behaviors. Throughout the survey period, the proportion of current users who binge changed very little and remained highest among persons aged 18-20 years (52.1%). CONCLUSIONS: Alcohol use leveled off in the 1990s, but may be increasing, especially among persons 18-20 years of age. Those who drink are about as likely to report binge drinking as were drinkers 15 years ago. PMID- 15110056 TI - Evaluation of field training in the epidemic intelligence service: publications and job choices. AB - BACKGROUND: Since 1951, the Centers for Disease Control and Prevention's Epidemic Intelligence Service (EIS) has provided training in applied epidemiology to physicians, nurses, veterinarians, dentists, and doctoral-level health scientists. About one third of these EIS officers have been trained in the setting of state and local health departments (the field). METHODS: To evaluate two specific outcomes of field EIS training, the authors reviewed the published work and career choices of field EIS officers after completing the program. The EIS classes of 1991-1996 were selected for study. A field officer was defined as an EIS Officer who completed at least the second year of a 2-year EIS assignment in a state or local health department position. RESULTS: During this period, 430 EIS officers completed the program; 117 (27.2%) were field officers. Of these, 84 (71.8%) published one or more scientific paper as first author for a total of 202 first authored manuscripts in over 50 different journals, an average of 1.7 (range, 0-8) per officer. Most (71%) were on infectious disease topics; 16% were on environmental health or injury control topics, and 11% were on chronic diseases. Field officers were more likely than headquarters-based officers to choose positions in state or local health departments for their first job after graduating (32/117 [27.4%] versus 22/313 [7.0%]; relative risk = 3.9, 95% confidence interval = 2.4-6.4). CONCLUSIONS: EIS training in the field has contributed to the scientific literature and to the strengthening of public health infrastructure at the state and local level. PMID- 15110057 TI - National availability of influenza vaccine among medical subspecialty practices. AB - BACKGROUND: Influenza vaccination rates fall short of national goals, particularly among individuals whose chronic conditions predispose them to complications of influenza. Availability of influenza vaccine in medical subspecialists' practices may affect vaccination rates among adults with chronic illness. METHODS: The practice sites of a national random sample of medical cardiology, endocrinology, and pulmonology physicians were contacted by telephone in February 2003 to March 2003 to determine which of them had influenza vaccine available to their patients during the 2002-2003 influenza season. The number of physicians in the practice and geographic location were also obtained. RESULTS: Office staff at the practices of 1683 of 2013 eligible physicians were successfully contacted, and 1473 provided information about vaccine availability. Overall, 1094 (74%) of practices had influenza vaccine available during the 2002 2003 season. Availability differed significantly by subspecialty: 54% cardiology, 78% endocrinology, and 90% pulmonology (p<0.001). Influenza vaccine was more often available at subspecialists' practices in the Northeast (80%) than in the South (74%), Midwest (71%), and West (70%; p<0.005). In multivariate analyses, pulmonology practices in all census regions and sizes were significantly more likely to have influenza vaccine available than was the reference cardiology practice. Several endocrinology practice types also had significantly higher influenza vaccine availability than those in cardiology practice, particularly in multi-physician practices. CONCLUSIONS: Influenza vaccine availability varies widely across practices in the three medical subspecialties that provide care to the largest numbers of individuals with an indication for the vaccine in the United States. These findings have implications for the accessibility of influenza vaccine to individuals at high risk for morbidity and mortality associated with influenza. PMID- 15110058 TI - Use of a state regulation for adult vaccination. AB - BACKGROUND: Influenza and pneumococcal disease are major causes of vaccine preventable death among the elderly. In an effort to raise immunization rates, New Jersey in 1999 adopted a regulation requiring hospitals to offer pneumococcal and influenza vaccinations to all inpatients aged 65 and over. This study examined the effect of implementation strategies on immunization rates within hospitals in 2000 and 2001. METHODS: Hospital infection control directors were surveyed and random chart review was conducted from a stratified sample of hospitals. The infection control director's assessment of their institution's success was a major outcome measure. RESULTS: Relatively few respondents thought their hospital has been successful in implementing new immunization protocols. Approximately 67% responded that they did not think physicians agreed with the "scope and nature" of the regulation. Physician attitudes led the list of barriers to implementation cited by respondents. Chart review revealed moderate amounts of immunization assessment performed by nurses, but virtually no evidence of physicians' orders for immunization. CONCLUSIONS: Thus far, there is little evidence that the New Jersey regulation has resulted in a meaningful change in pneumococcal or influenza vaccination practices. PMID- 15110059 TI - Physicians' roles in preventing dental caries in preschool children: a summary of the evidence for the U.S. Preventive Services Task Force. AB - CONTEXT: Almost 20% of children aged 2 to 5 years have untreated dental caries. Physician interventions to prevent and manage dental caries in preschool children could help address this common problem. OBJECTIVE: To review the evidence for effectiveness of five possible physician interventions- (1) screening and risk assessment, (2) referral, (3) provision of dietary supplemental fluoride, (4) application of fluoride varnish, and (5) counseling-for the prevention of dental caries for the U.S. Preventive Services Task Force. DATA SOURCES: Articles from 1966 to 2001 addressing the effectiveness of primary care clinicians' interventions to prevent or manage dental caries were identified in MEDLINE. The evidence for effectiveness of supplemental fluorides, fluoride varnish, and counseling for caries prevention performed by dental personnel was also examined through existing and new systematic reviews. DATA SYNTHESIS: For most key questions related to the five interventions, the evidence for primary care clinician effectiveness was rated as poor owing to the scarcity of studies. Ten surveys of physicians' knowledge and behavior about fluoride supplementation provided fair evidence, suggesting that supplementation decisions were often made without consideration of other fluoride exposures. Reviews of the dental literature identified fair evidence supporting the effectiveness of both fluoride supplements and varnish, although information describing effectiveness and adverse outcomes of supplementation with the most recent dosage schedule is not available. CONCLUSIONS: Evidence for the effectiveness of traditionally recommended primary care clinician interventions (screening, referral, counseling) to prevent dental caries in preschool children is lacking. There is fair evidence for the effectiveness of two fluoride-based interventions (fluoride supplementation and varnish) applicable in primary care practice. However, there is also fair evidence indicating that physicians' consideration of fluoride exposure is incomplete, thus increasing the risk for fluorosis among those prescribed supplements. PMID- 15110060 TI - Prevention of dental caries in preschool children: recommendations and rationale. PMID- 15110061 TI - Stage-based lifestyle interventions in primary care: are they effective? AB - BACKGROUND: To systematically review the literature concerning the effect of stages-of-change-based interventions in primary care on smoking, physical activity, and dietary behavior. METHODS: An extensive search (until July 2002) was performed using the following inclusion criteria: (1) (randomized) controlled trial (RCT/CT), (2) intervention initiated in primary care, (3) and intervention aimed at changing smoking, physical activity, or dietary behavior, and stages-of change-based outcomes, and (4) behavioral outcomes. Methodologic quality was assessed, and conclusions on the effectiveness at short-, medium-, and long-term follow-up were based on a rating system of five levels of evidence. Odds ratios were calculated when methodologically appropriate. RESULTS: A total of 29 trials were selected for inclusion. Thirteen studies included a physical activity intervention, 14 aimed at smoking cessation, and five included a dietary intervention. Overall methodologic quality was good. No evidence was found for an effect on stages of change and actual levels of physical activity. Based on the strength of the evidence, limited to no evidence was found for an effect on stages of change for smoking and smoking quit rates. Odds ratios for quitting smoking showed a positive trend. Strong evidence was found for an effect on fat intake at short- and long-term follow-up. Limited evidence was found for an effect on stages of change for fat intake at short-term follow-up. CONCLUSIONS: The scientific evidence for the effect of stages-of-change-based lifestyle interventions in primary care is limited. Limiting aspects in the stages-of change concept with respect to complex behaviors as physical activity and dietary behavior are discussed. PMID- 15110062 TI - Expanding developmental and behavioral services for newborns in primary care: program design, delivery, and evaluation framework. AB - BACKGROUND: Healthy Steps (HS) for Young Children strengthens the healthcare system as a source of developmental and behavioral support for parents. This series of papers presents a study of HS as implemented within a large health maintenance organization that tested the benefit of beginning intervention services during pregnancy with an extension program called "PrePare" (PP). METHODS: The design was a quasi-experimental comparison of intervention families with families receiving usual care. Within the intervention, families were assigned randomly to begin receiving Healthy Steps services prenatally (PP+HS) or shortly after birth (HS). We used a systems model, PRECEDE/PROCEED, for planning, implementation, and process evaluation. Outcomes examined when the infants were aged 3 months included changes in family social support and capacity for parenting, parenting behaviors, and satisfaction and loyalty to the health plan. RESULTS: The sample of 439 families was distributed as follows: usual care (n=136), prenatal initiation of services (PP+HS; n=151), and postnatal Healthy Steps (HS; n=152). Information about program implementation, including provider satisfaction, is provided for the early phases of the study (through age 3 months). The intervention was delivered with fidelity and with minimal disruption to the practice styles of pregnancy providers, most of whom considered the program valuable to their patients. Relative to families in the comparison group, families in the intervention group received more usual care services and more intervention-specific services. CONCLUSIONS: The pregnancy and newborn phases of the intervention were embedded successfully within the existing healthcare delivery system. The program was considered valuable for parents by providers and parents. Participating families received more services and a greater variety of services than families in usual care. Whether these differences result in beneficial outcomes for families or the health plan are topics of the subsequent papers. PMID- 15110063 TI - Expanding developmental and behavioral services for newborns in primary care; Effects on parental well-being, practice, and satisfaction. AB - BACKGROUND: Healthy Steps (HS) was designed to address, prospectively, behavioral and developmental support needs of young families in pediatric clinical care settings. PrePare (PP) initiates these services prenatally, whereas HS begins services in the postnatal period. Both interventions have universal and risk directed components. Intervention effects in the first 3 months after birth are reported here. METHODS: A quasi-experimental design was used to allocate 439 participants to intervention or usual care conditions. Within the intervention group, enrollees were randomly assigned to receive HS or PP+HS services. Early outcomes were assessed by telephone survey at 1 week and 3 months postpartum. RESULTS: Mothers in either intervention condition were less likely to report depressive symptoms and more likely to describe themselves as pleased in their role as parents. Intervention families were more likely to continue breastfeeding and more likely to read to their 3-month-old. Knowledge of infant development and recognition of appropriate discipline was greater among intervention recipients. Satisfaction with pediatric care was higher among intervention recipients and the rate of health plan disenrollment was 75% lower at 3 months among those enrolled in the prenatal intervention. No other outcome difference emerged between HS and PP+HS enrollees. CONCLUSIONS: Receipt of either intervention was associated with positive effects on health, safety, and developmentally appropriate parenting, as assessed in early infancy. There were positive effects on health plan disenrollment. No additional benefit could be ascribed to prenatal institution of services. A combination of universal and risk-based support for new parents is recommended, rather than the provision of risk-based services alone. PMID- 15110064 TI - Expanding developmental and behavioral services for newborns in primary care: implications of the findings. AB - BACKGROUND: In two other papers in this issue, the rationale, development, implementation, experimental design, approach to evaluation, and early results of a program to deliver developmental and behavioral services to all infants in primary care practice were described. Positive effects were seen for parental satisfaction, including decreased disenrollment, provider satisfaction, parenting practices, and health outcomes. METHODS: In the present article, the results are reviewed and implications of our findings for the delivery of care, families, healthcare systems, and further research are discussed. RESULTS: Findings that have broad implications are as follows: (1) developmental and behavioral services can be delivered successfully in practice using dedicated professionals to deliver and integrate services; (2) the "planned care model" was useful in program implementation for making "the right thing to do, the easy thing to do"; (3) the added focus on satisfaction and cost helps to develop the "business case" for broad scale implementation; (4) bonding of parents to organizations has marketing implications; (5) the program provides positive effects for all parents, not just high-risk parents; and (6) several research questions emerge, including persistence of effects on health outcomes, costs, and utilization. CONCLUSIONS: The authors conclude that study results have implications for preventive services, families, child healthcare in office practice, healthcare systems, and healthcare policy. In this ongoing study, examination of intervention effects at 30 months of age shoud be informative. Further research is warranted as it remains to be seen whether or not these interventions can become viable ongoing programs. PMID- 15110065 TI - Optimizing vaccine availability and utilization: position statement of the American College of Preventive Medicine. PMID- 15110066 TI - The camel's nose is under the tent: opportunities for prevention associated with the 2003 Medicare Act. PMID- 15110067 TI - Diagnostic efficacy of biochemical markers in diagnosis post-mortem of ischaemic heart disease. AB - In forensic medicine, there is a need for more sensitive biochemical markers for the post-mortem diagnosis of acute myocardial infarction. A study of the distribution of biochemical markers in different fluids is of great significance in post-mortem diagnosis, because their distribution depends on the location of tissue damage and release kinetics. The aim of this study is to compare the sensitivities and specificities of creatine kinase-MB (CK-MB), myoglobin and cTnI in serum and pericardial fluid for the post-mortem diagnosis of acute myocardial infarction (AMI). We studied 188 cadavers selected during 1 year from medicolegal autopsies. The groups were as follows: (1) myocardial infarction (n = 52); (2) asphyxia (n = 59); (3) multiple trauma (n = 41); (4) natural deaths excluding myocardial infarction (n = 36). We obtained statistically significant differences in pericardial fluid for all the biochemical markers, the highest levels being obtained in the group of cadavers who had died from myocardial infarction. A common factor is the high negative predictive value found in biochemical markers, which is contrary to the findings obtained in clinical practice, when the percentages of sensitivity are very high. PMID- 15110068 TI - Rapid determination of ketamine in urine by liquid chromatography-tandem mass spectrometry for a high throughput laboratory. AB - In recent years, the abuse of ketamine had gained popularity in rave parties in Hong Kong. The Urinalysis Unit of the Government Laboratory of Hong Kong faced a tremendous increase in workload for ketamine analysis. The number of tests performed rose from 10 in 1999 to 15,000 in 2002. As a fully validated immunoassay test for ketamine was not available in the market, most laboratories analyzed ketamine by chromatographic techniques after liquid-liquid extraction. However, these methods reported in the literature are not suitable for high throughput laboratories. Hence, a rapid screening/confirmation method for ketamine by liquid chromatography-tandem mass spectrometry (LC-MS-MS) with a detection limit of 5 ng/ml was developed. After automated solid-phase extraction (SPE), the urine extract was analyzed for ketamine by a 2.5-min chromatographic run, the estimated recovery was 89% and the precision was 11% R.S.D. at 20 ng/ml. With the aid of an in-house developed computer program, the results were presented in spreadsheet format for easy checking. The method has been applied to our laboratory for routine ketamine analysis and a maximum of 200 samples per day can be achieved. PMID- 15110069 TI - Fatal and non-fatal injuries caused by crossbows. AB - Today in modern times, traumatic injuries caused by crossbows are a rarity. The largest collection of cases so far is presented in this study, consisting of four fatalities (two homicides and two suicides) and two non-fatal injuries (grievous bodily harm and an accident). All the victims were male having an age between 31 and 54. The weapons, which were used, were mainly high-performance precision crossbows with telescopic sights and hunting bolts. The parts of the body involved were the facial/head area in three of the cases and the thorax in three of them. There were either deep or total penetration injuries to the cranium and thorax with the bolt remaining in the wound in four out of six cases. The persons with non-fatal crossbow injuries exhibited comparatively few symptoms, despite the sometimes extensive involvement of the interior of the cranium (cerebrocranial penetration, in one instance). The two cases of suicide favoured the body areas often found with gun-users. The aetiological classification of crossbow injuries may be difficult after the removal of the bolt. The external morphology is strongly dependent on the type of tip used. Multiple-bladed hunting broadheads produce radiating incised wounds, whereas conical field tips produce circular to slitlike defects. Correspondingly, the external injuries can be reminiscent of the effects of a violent attack by sharp force or of a gunshot wound. The possibility, supported by clinical data, that the victim might have the ability to act or even to survive for a period of time, even with penetration of the brain, should be taken into account when the cause of death is being investigated. PMID- 15110070 TI - Suicidal deaths in childhood and adolescence. AB - Suicide has been reported as the second or third most common cause of death in children and adolescents worldwide. In this study, cases of under the age of 19 years submitted to the Institute of Forensic Medicine, First Specialization Board between 1996 and 2000 as suicides by the Board were evaluated retrospectively. The cases included in this study were the cases bearing locally questionable components, so had been submitted to the evaluations by the Board in order to eradicate the doubts. A total of 43 cases were investigated regarding age, gender, cause of death, manner of death, place of death, time of death, and the risk factors. Of the 43 cases evaluated, 31 cases were female and 12 cases were male. The notable suicide method was found to be firearms. Although it was clear that that not all of the suicide cases in this age group had been submitted to the Institute of Forensic Medicine, the most striking result of this study, nevertheless, was that girls constituted the 72% of suicidal deaths in this age group. PMID- 15110071 TI - An unusual suicide with a safety razor blade--a case report. AB - A 29-year-old male was found dead lying in a pool of blood inside a community toilet locked from inside. The individual is alleged to have committed suicide by cutting his throat with a safety razor blade, which was found at the scene by the investigating police authority. The deceased, as per his relatives, was suffering from depression for the last few days. PMID- 15110072 TI - Analysis of 16 Y STR loci in the Finnish population reveals a local reduction in the diversity of male lineages. AB - We analysed samples of 400 Finnish males using nine Y-chromosomal short tandem repeat (STR) loci (minimal haplotype); for 200 of these subjects an additional seven Y-chromosomal STR loci were used. The geographical distribution of the observed haplotypes was determined from 200 individuals of known paternal origin within Finland. The observed number of alleles varied from 2 to 13 alleles per locus. A total of 146 minimal haplotypes were identified in our population sample. Interestingly, 90 (22.5%) individuals shared an identical haplotype. This haplotype was extremely frequent in the northern and eastern subpopulations of Savo, Pohjanmaa and Karjala (53, 42 and 37%, respectively). With the seven additional loci analysed in the sample of 200 individuals, 120 haplotypes were identified, and individuals sharing the most common haplotype decreased to 13.0%. However, in comparison to other European populations, the Finnish population showed decreased genetic diversity (GD) when the number of different minimal haplotypes in the population was divided by the sample size (36.5% in Finns versus 83.7% on average). Our results strongly support the earlier hypothesis of individual isolated Y-chromosomal lineages and population substructuring in Finland. For paternity testing, power of exclusion was 92% using minimal haplotype data, but including the seven additional loci this value increased to 97%. PMID- 15110073 TI - Y-chromosome STR-haplotype typing in El Salvador. AB - Eight Y-chromosome STRs were investigated in a male population sample from El Salvador. Complete Y-chromosomal STRs haplotypes were obtained in 121 individuals, among which 107 different haplotypes were observed. The two most common haplotypes were shared by approximately 4% of the sample, while 100 haplotypes were unique. The gene diversity was 0.9883 and the discrimination capacity was 0.8926. The combined Y-chromosome STR polymorphisms provide a powerful discrimination tool for routine forensic applications. PMID- 15110074 TI - Genetic diversity and admixture data on 11 STRs (F13B, TPOX, CSF1PO, F13A01, D7S820, LPL, TH01, vWA, D13S317, FESFPS, and D16S539) in a sample of Rio de Janeiro European-descendants population, Brazil. AB - Allelic frequencies, forensic parameters and admixture values for eleven STR loci (F13B, TPOX, CSF1PO, F13A01, D7S820, LPL, TH01, vWA, D13S317, FESFPS, and D16S539) were determined in a sample of unrelated individuals, European descendants from Rio de Janeiro area, Brazil. PMID- 15110075 TI - STR data for the AmpFlSTR SGM Plus from the eastern and western sections of Mediterranean region of Turkey. AB - Allele frequencies of the 10 STR loci (D16S539, D2S1338, D3S1358, vWA, D18S51, D21S11, D8S1179, D19S433, FGA and TH01) included in the AmpFlSTR SGM Plus kit were determined in 107 unrelated individuals from the eastern section and 108 unrelated individuals from western section of Mediterranean region of Turkey. The expected performance of these loci for personal identification and paternity testing in these populations were estimated. PMID- 15110076 TI - Immunotherapeutic effects of T11TS/S-LFA3 against nitrosocompound mediated neural genotoxicity. AB - Nitrosocompounds formed by the interaction of nitrites and secondary amines are neurotoxic in human and different rodent species. Human exposure of nitrosocompounds are widespread affected by different modes like nitrite/nitrate preserved foods, beverages like beer, formed in the stomach following uptake of the precursors nitrates, nitrites and secondary amines. The productions of alkylating metabolites during the breakdown of nitrosocompounds are the causative agents for the neurotoxic changes of the neural cells. An attempt has been made in our lab to study the effect of nitrosocompound mediated toxicity and the gradual toxic effects of these neurotoxic agents to transform the normal glial cells to a neoplastic one. The present study indicated that a transmembrane glycopeptide of sheep red blood cell (SRBC), known as S-LFA3 or T11 target structure (T11TS) applied to nitrosocompound induced animals manifesting a full grown intracranial malignancy can revert back tumor-bearing condition to the normal physiological state. Young Druckray rat of both sexes aging 3-5 days were injected with N'-N'-ethyl nitrosourea (ENU) intraperitoneally (i.p.) at a single dose of 80 mg/kg body weight to simulate nitrosocompound mediated neurotoxicity. 2-,4-, 6-, 8-, and 10-month-old neonatal ENU induced animals were sacrificed for growth kinetics, functional immunological parameters and receptor studies to hint at the changes during tumor development. In order to determine the immunomodulatory role of T11TS, 7-month-old ENU induced animals were injected with T11TS at a dose of 0.41 mg/kg body weight, in three consecutive doses at an interval of 6 days maintaining normal control as untreated control and ENU induced animals of age-matched rats as tumor-bearing control. All the immunological parameters, growth kinetic study, receptor-based study by FACS directly established the immunomodulatory, anti-toxic and anti-tumor property of T11TS/S-LFA3. Finally, formation of DNA ladder along with the FACS-based apoptosis study clearly indicated that T11TS is a potent apoptotic inducer in neoplastic neural cells. PMID- 15110077 TI - Attenuation of paraquat-induced motor behavior and neurochemical disturbances by L-valine in vivo. AB - Alterations of motor behavioral patterns and monoamine contents in the discrete rat brain areas after acute paraquat exposure (3, 5, 10, 20 mg/kg, s.c.) have been studied. The results showed that paraquat at the doses of 5, 10, and 20 mg/kg significantly reduced locomotive, stereotypic, and rotational behaviors. Significant decreases of norepinephrine (NE) contents in cortex and hypothalamus, as well as striatal contents of dopamine (DA) and its acidic metabolites, were detected. In addition, L-valine (200 mg/kg, i.p.) significantly attenuated paraquat-induced toxicity at moderate dose (5 mg/kg) but not at high dose (20 mg/kg). The results provide evidence that paraquat can enter the brain as illustrated by the alterations in the motor behavioral pattern and neurochemical contents. Furthermore, the attenuation effect of L-valine against systemic administration of paraquat-induced motor behaviors was detected, with a slightly protective effect on paraquat-induced neurochemical alterations. PMID- 15110078 TI - Concomitant administration of an isopropanolic extract of black cohosh and tamoxifen in the in vivo tumor model of implanted RUCA-I rat endometrial adenocarcinoma cells. AB - Black cohosh is a well known herbal remedy of long traditional use against menopausal complaints. Recently published studies on postmenopausal hormone replacement with synthetic substances associated severe negative side effects with an increase in duration of administration. The subsequent popularity of alternative treatments, often herbal drugs, made investigations into the safety of these preparations more pressing. Until now, black cohosh demonstrated no estrogen-agonistic activity in mammary cells, neither in animal model nor in cell culture, i.e., no gene transcription or cell proliferation was induced. Here we tested for the influence of a standardized isopropanolic extract of black cohosh on an animal model of endometrial cancer. Ectopic growth of the primary tumor as well as the incidence and localization of metastases were examined, partly in the setting of a combination treatment with tamoxifen. In contrast to the endometrial estrogen agonist tamoxifen, black cohosh did not further growth or metastasizing potential of the primary tumor. Absence of detectable supportive or antagonistic effects between both treatments most probable come from the relatively high tamoxifen dose. PMID- 15110079 TI - Effect of two nitrogenous diphenyl ether pesticides on mast cell activation. AB - We examined the effect of two nitrogenous diphenyl ether pesticides, nitrofen (NIP) and chlornitrofen (CNP), on mast cell activation. RBL-2H3 (rat basophilic leukemia) cells were exposed to NIP or CNP for 30 min to investigate their effect on degranulation, and for 3 h to investigate their effect on cytokine production and gene expression. NIP and CNP increased IgE receptor-mediated beta hexosaminidase release, MCP-1 release, and TNF-alpha release in a dose-dependent manner. The increasing effect of CNP on their release was greater than that of NIP. In the gene expression experiment, 30 microg/ml CNP significantly upregulated Egr-1, MCP-1 and GADD45a gene expression. These results suggest that at higher concentrations (more than 30 microg/ml) the nitrogenous diphenyl ether pesticides had both a degranulation-enhancing effect and proinflammatory cytokine production enhancing effect through the expression of some transcription factors in RBL-2H3 cells. PMID- 15110080 TI - Effects of epidermal growth factor receptor deficiency and 2,3,7,8 tetrachlorodibenzo-p-dioxin on fetal development in mice. AB - Dioxins are persistent environmental contaminants that cause multiple disorders in laboratory animals, including teratogenesis. In mice, the most important teratogenic effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) are hydronephrosis and cleft palate. Aryl hydrocarbon receptor (AHR) mediates most of the TCDD-induced effects, but modulation of these effects by other factors such as epidermal growth factor receptor (EGFR) has been propounded. TCDD changes the expression of both EGF and its receptor EGFR, which may be one step in the pathway leading to cleft palate and hydronephrosis. In the present study, the importance of EGFR in TCDD-induced teratogenicity was evaluated. Heterozygous EGFR(+/-)-mice were mated and pregnant females exposed to 1.5-106.0 microg/kg TCDD on gestation day (GD) 10 and killed on GD 18. The fetuses were studied for cleft palate, hydronephrosis, and open eyes. There was no marked difference among the three genotypes in sensitivity to cleft palate or hydronephrosis, but in EGFR(-/-)-mice frequency of the open eye malformation decreased dose-dependently. In conclusion, EGFR signaling is not required for TCDD-induced cleft palate or hydronephrosis but TCDD appears to counteract the effect of EGFR deficiency on eye opening. PMID- 15110082 TI - Gene expression changes in peripheral blood-derived dendritic cells following exposure to a contact allergen. AB - A critical step in the induction of allergic contact allergy is the activation and subsequent migration of Langerhans cells (LC), an important antigen presenting dendritic cell (DC) of the skin. As the Langerhans cells migrate, they undergo a maturation process. It has been proposed that contact allergen exposure can induce DC maturation. While changes in DC gene expression profiles induced by various maturation stimuli have been explored, there are no published reports describing genomic-scale analysis of the changes induced by chemical allergen exposure. Therefore, to explore the concept of chemical allergen-induced DC maturation and to identify genes that are regulated by exposure to allergens we examined, at the transcriptional level, the effects of exposure to a contact allergen on DC. Peripheral blood-derived DC were exposed for 24 h to either 1mM or 5 mM dinitrobenzenesulfonic acid (DNBS). Changes in gene expression were analyzed using Affymetrix U95Av2 GeneChip. Comparison of mean signal values from replicate cultures revealed 173 genes that were significantly different (P < or = 0.001) between 1 mM DNBS treated and untreated control DC and 1249 significant gene changes between 5 mM DNBS treated and control DC. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) was used to evaluate the observed transcript changes for selected genes in DC derived from a second donor. Comparison of the fold-changes in transcript levels between the two platforms and donors revealed a good correlation in both direction and magnitude. RT-PCR analysis was also used to assess the allergen specificity of a selected number of genes in DC derived from a third donor. Many of the gene expression changes were found to be induced only by exposure to the allergen, DNBS, and not by exposure to a structurally similar non-allergen, benzenesulfonic acid. A number of gene expression changes induced by allergen exposure were found to be consistent with what is known of the DC maturation process, and thus provide support for the theory of contact allergen-induced DC maturation. Additionally, it is hoped that some of the transcript changes identified through this approach will be shown to be suitable for use in the development of an in vitro predictive assay for contact sensitization. PMID- 15110081 TI - The dioxin-like pollutant PCB 126 (3,3',4,4',5-pentachlorobiphenyl) affects risk factors for cardiovascular disease in female rats. AB - Epidemiological studies suggest that exposure to persistent organic pollutants such as organochlorines might induce cardiovascular disorders and diabetes. Some of these organochlorines, such as dioxins and some dioxin-like PCBs, have been characterised as anti-estrogenic due to their inhibition of estrogenic-induced responses. In the present pilot study, 40 female rats were subjected to either exposure to the dioxin-like 3,3',4,4',5-pentachlorobiphenyl (PCB 126) or vehicle, as well as ovariectomy (OVX) or sham operation in a 2 x 2 factorial design over 12 weeks to explore potential interactions between estrogen status and PCB 126 exposure on cardiovascular risk factors. PCB 126 increased heart weight and serum cholesterol levels in both groups. PCB 126 increased blood pressure in the sham operated animals only. In conclusion, PCB 126 exposure in female rats resulted in effects on cardiovascular risk factors, such as serum cholesterol, blood pressure, and heart weight. Of these effects of PCB 126, the increase in blood pressure was dependent on estrogen status. PMID- 15110083 TI - Cell-specific oxidative DNA damage induced by estrogen in rat testicular cells in vitro. AB - 17 alpha-Ethinylestradiol (EE) can induce oxidative DNA damage in terms of 8-oxo 7,8-dihydro-2'-deoxyguanosine (8-oxodG) in rat testicular cells by an apparent estrogen receptor-mediated mechanism. We investigated differential susceptibility to EE in cell sub-populations from rat testes and the role of rat 8-oxo-guanine DNA glycosylase (rOGG1). Isolated rat testicular cells were incubated with EE concentrations ranging from 0.1 to 1000 nM. Single strand DNA breaks and oxidised purines as fapyguanine glycosylase (FPG) sensitive sites were assessed by the comet assay. In the total cell population and in round haploid cells, oxidised purines showed a bell-shaped concentration-response relationship with a maximally increased levels at 10 nM EE, whereas, no significant effects were seen in diploid, S-phase or tetraploid cells. The mRNA level of rOGG1 in testes cells was unaffected by EE, whereas, baseline levels were higher than in liver tissue and similar to colon tissue. PMID- 15110084 TI - Detection of Purkinje cell loss following drug exposures to developing rat pups using reverse transcriptase-polymerase chain reaction (RT-PCR) analysis for calbindin-D28k mRNA expression. AB - A technique is described that allows for the identification and quantification of Purkinje cell loss in cerebellum subsequent to developmental toxic exposures. This technique relies upon the extensively validated findings that the Purkinje cell is the only site of expression in the cerebellum of the calcium binding protein calbindin-D28k. Thus, analysis of mRNA expression specific to this protein by comparison to matched controls provides a reliable means of determining whether cell loss has occurred. Purkinje cell loss was induced in rat pups by ethanol exposure on postnatal day (PN) 4 or valproic acid administration to pregnant dams on gestational day 13. Analysis was conducted on PN5 or PN10 and the results compared to parallel groups of pups where the Purkinje cells were counted by traditional means. When compared to matched control rat pups the decrease in calbindin-D28k mRNA expression indicates Purkinje cell loss regardless of whether the cell loss was induced by prenatal valproic acid or postnatal ethanol exposure. The availability of a biochemical alternative to histological cell counting allows for more detailed analyses of the mechanisms of Purkinje cell death induced by these two toxicants, including analyses of the early alterations in signal transduction proteins. PMID- 15110085 TI - Comparison of the susceptibility of wild-type and CYP2E1 knockout mice to the hepatotoxic and pneumotoxic effects of styrene and styrene oxide. AB - Styrene causes both liver and lung damage in non-Swiss albino, CD-1, and other strains of mice. This is considered to be due to the bioactivation of styrene to styrene oxide by cytochromes P450, principally CYP2E1 and CYP2F2. If so, one would expect CYP2E1 knockout mice to be less susceptible to styrene-induced toxicity than wild-type mice. However, previous in vitro and in vivo studies demonstrated little difference in the metabolism of styrene to styrene oxide between wild-type and CYP2E1 knockout mice. These findings would suggest that there should be no difference in the toxic responses to styrene between these two strains. To determine which of these possibilities was correct, styrene (600 mg/kg) or styrene oxide (300 mg/kg) was administered i.p. 24 h prior to measurement of serum sorbitol dehydrogenase as a biomarker of hepatotoxicity or lactate dehydrogenase activity, protein, and cells in bronchoalveolar lavage fluid as biomarkers for pneumotoxicity. Styrene was more hepatotoxic in the wild type mice than in the knockout mice suggesting CYP2E1 activity is important. Strain differences were not observed with styrene oxide indicating no difference in intrinsic susceptibility. For lung, the response was similar in both strains to both styrene and styrene oxide supporting the idea that CYP2F2 is important in the bioactivation of styrene in this tissue and that there is no strain difference in susceptibility to the active metabolite. PMID- 15110086 TI - Increased effects of MPDAX, a novel xanthine derivative, on antitumor activity of doxorubicin. AB - To clarify the effect of 1-methyl-3-propyl-7-N,N-dimethylpropylamide-xanthine (MPDAX) on doxorubicin (DOX) transport, we examined the efficacy of MPDAX as an amplifier of the antitumor activity of DOX in mice bearing tumors with different properties as to DOX transport across cell membranes. MPDAX significantly enhanced the DOX-induced antitumor activity on DOX-sensitive tumors. It is expected that the increase in antitumor activity caused by MPDAX contributes to the increased DOX concentration in tumors due to the MPDAX-induced change in DOX transport via the transporter expressed in sensitive tumor cells. In contrast, in M5076, a lower sensitive to DOX, MPDAX decreased the tumor weight by half at an otherwise ineffective dose of DOX. Furthermore, in P388/DOX, DOX has no effect, but MPDAX caused an elevation of the DOX-induced antitumor activity with an increase in the DOX concentration in the tumors. The results suggested that MPDAX is a novel amplifier for antitumor agents as it significantly increased the antitumor activity toward tumors with different properties. The DOX concentrations in the MPDAX + DOX group for all tumor lines were about two-fold those in the DOX alone group. Furthermore, MPDAX and DOX exhibited significant inhibitory effects on uridine and thymidine uptake. It is known that nucleoside transporters increase the membrane permeability of DOX. We speculated that MPDAX inhibits the cell membrane transport of uridine and thymidine via nucleoside transporters. MPDAX, acting via nucleoside transporters, increases the DOX induced antitumor activity toward many tumor types and is an useful biochemical modulator. PMID- 15110087 TI - Comparative mixture effects of JP-8(100) additives on the dermal absorption and disposition of jet fuel hydrocarbons in different membrane model systems. AB - Jet fuel are complex mixtures of hydrocarbon fuel components and performance additives. Three different membrane systems, silastic, porcine skin and the isolated perfused porcine skin flap (IPPSF) were used to gain insight into the possible mechanism for additive interactions on hydrocarbon component absorption. Influence of JP-8(100) additives on the dermal kinetics of 14C-naphthalene and 14C/3H-dodecane as markers of hydrocarbon absorption, were evaluated using analysis of means (ANOM) and analysis of variance (ANOVA). This study indicated that the naphthalene absorption through silastic membrane was significantly different with JP-8 plus individual additives as compared to controls, i.e. JP-8 and JP-8(100). The porcine skin data indicated that neither individual nor combinations of additives affected naphthalene absorption. The third membrane system (IPPSF) showed that only MDA and BHT were important additives altering naphthalene absorption. MDA was a significant suppressor while BHT was a significant enhancer of naphthalene absorption. MDA significantly decreased dodecane absorption in skin flaps. All individual and combinations of two additives with JP-8 affected naphthalene and dodecane surface retention in silastic membrane. The IPPSF indicated that only 8Q405 is a significant modulator of surface retention for both marker hydrocarbons. The 8Q405 significantly reduced naphthalene contents in dosed silastic and skin indicating a direct interaction between additive and marker hydrocarbons. The MDA and BHT, which significantly retained naphthalene in the stratum corneum of porcine skin individually, led to a statistical decrease in its retention in the stratum corneum when in combination (MDA + BHT) suggesting a potential biological interaction. These observations demonstrate that the single membrane system may not be suitable for the final prediction of complex additive interactions in jet fuels. Rather a combination of different membrane systems may provide the insight to elucidate the possible mechanism for additive interactions. Finally, it is important to assess all components of a chemical mixture since the effects of single components administered alone or as pairs may be confounded when all are present in the complete mixture. PMID- 15110088 TI - Effects of dietary di(2-ethylhexyl)phthalate, a putative endocrine disrupter, on enzyme activities involved in the metabolism of tryptophan to niacin in rats. AB - We have reported that the conversion ratio of tryptophan to niacin increased with increasing dietary concentration of di(2-ethylhexyl)phthalate (DEHP); the conversion ratio was about 2.0% in the control rat, which increased by about 30% in the rat fed with 3.0% DEHP diet. In this study, we investigated whether this abnormal increase in the conversion ratio by DEHP occurred through the alteration of the enzyme activities involved in the metabolism of tryptophan to niacin. Rats were fed with a diet containing 0%, 0.1%, 0.5%, or 1.0% DEHP for 21 days. The nine kinds of enzyme activities involved in the biosynthesis and catabolism in the liver and kidney were measured. Based on previous findings that the formation of quinolinic acid and its' metabolites significantly increased with DEHP administration, we proposed that the activity of 2-amino-3-carboxymuconate-6 semialdehyde decarboxylase would be inhibited by DEHP intake. However, we found that the activities in the liver and kidney did not decrease in the rat fed with DEHP-containing diet. We discuss the discrepancy between the metabolite results and the enzyme activities. PMID- 15110089 TI - Proteolytic fragments of ovalbumin display antimicrobial activity. AB - Ovalbumin, one of the major proteins present in avian egg white, was proteolytically digested by trypsin and chymotrypsin and the peptide fragments were investigated for their antimicrobial activity. The antimicrobial peptides were isolated and characterized. From the tryptic digestion, the following five antimicrobial peptide fragments were obtained: SALAM (residues 36-40), SALAMVY (residues 36-42) YPILPEYLQ (residues 111-119), ELINSW (residues 143-148) and NVLQPSS (residues 159-165). Digestion of ovalbumin by chymotrypsin yielded the antimicrobial peptides AEERYPILPEYL (residues 127-138), GIIRN (residues 155-159) and TSSNVMEER (residues 268-276). The peptides were synthesized and found to exert antimicrobial activity. They were strongly active against Bacillus subtilis and to a lesser extent against the other bacterial strains examined. A weak fungicidal activity against Candida albicans was also shown by some peptides. Ovalbumin itself was not bactericidal against all the bacteria strains examined. Our results suggest that the food protein ovalbumin may supply the organism with antimicrobial peptides, supporting the immunodefences of the organism. PMID- 15110090 TI - Functional co-expression of xenobiotic metabolizing enzymes, rat cytochrome P450 1A1 and UDP-glucuronosyltransferase 1A6, in yeast microsomes. AB - Xenobiotic Phase I and Phase II reactions in hepatocytes occur sequentially and cooperatively during the metabolism of various chemical compounds including drugs. In order to investigate the sequential metabolism of 7-ethoxycoumarin (7EC) as model substrate in vitro, xenobiotic metabolizing enzymes, rat cytochrome P450 1A1 (P450 1A1) and UDP-glucuronosyltransferase 1A6 (UGT1A6) were co-expressed in Saccharomyces cerevisiae AH22. Rat P450 1A1 and yeast NADPH-P450 reductase were expressed on a multicopy plasmid (pGYR1) in the yeast. Rat UGT1A6 cDNA with a yeast alcohol dehydrogenase I promoter and terminator was integrated into yeast chromosomal DNA to achieve the stable expression. Co-expression of P450 1A1 and UGT1A6 in yeast microsomes was confirmed by immunoblot analysis. Protease treatment of the microsomes showed the correct topological orientation of UGT to the membranes. The metabolism of 7EC to 7-hydroxycoumarin (7HC) and its glucuronide in yeast microsomes was analyzed by reverse phase HPLC. In a co expression system containing 7EC, NADPH and UDP-glucuronic acid, glucuronide formation was detected after a lag phase, following the accumulation of 7HC. In the case of P450 1A1 and UGT1A6, efficient coupling of hydroxylation and glucuronidation in 7EC metabolism was not observed in the co-expression system. This P450 and UGT co-expression system in yeast allows the sequential biotransformation of xenobiotics to be simulated in vitro. PMID- 15110091 TI - Structural characteristic of terminal dicarboxylic moiety required for apoptogenic activity of alpha-tocopheryl esters. AB - alpha-Tocopheryl succinate (TS) is known to induce apoptosis in various cells and has attracted attention as a chemotherapeutic agent. Recently, we reported the structural significance of the terminal dicarboxylic moiety for the action of TS [J. Nutr. Sci. Vitaminol. 49 (2003) 310-314]. In this study, to determine details of the relationship between the structure and the function of the terminal ester moiety of alpha-tocopherol (alpha-T), we synthesized four novel esters, alpha tocopheryl oxalate (TO), alpha-tocopheryl malonate (TM), alpha-tocopheryl pimelate (TP) and alpha-tocopheryl succinate ethyl ester (TSE), and compared their apoptogenic activities with those of TS, alpha-T, gamma-tocopherol (gamma T) and two commercially available alpha-T derivatives, alpha-tocopheryl nicotinate (TN) and alpha-tocopheryl acetate (TA), in vascular smooth muscle cells and a mouse breast cancer cell line C127I. TO and TM in addition to TS, but not the others, induced apoptosis in both cells. Particularly, TO was the most potent of all alpha-T derivatives used. The addition of exogenous superoxide dismutase (SOD) significantly prevented the apoptosis induced by TM as well as that by TS as reported previously, but did not affect TO-induced apoptosis. These results suggest that O(2)(-) generated exogenously participates in TM-induced apoptosis but not in TO-induced apoptosis. The difference in their apoptotic effects is attributed to structural properties of the terminal dicarboxylic moiety, which has an inflexible plane conformation in TO, while it is highly flexible in TM and TS. PMID- 15110092 TI - Effect of chlorophyllin against oxidative stress in splenic lymphocytes in vitro and in vivo. AB - Chlorophyllin (CHL) has been examined as an antioxidant/radioprotector in splenic lymphocytes from BALB/c mice. CHL inhibited lipid peroxidation induced by 2,2' azobis(2-propionimidinedihydrochloride) (AAPH) in lymphocytes in vitro. It also partially prevented radiation-induced suppression of mitogenic stimulation of lymphocytes in vitro. Generation of intracellular reactive oxygen species (ROS) by radiation or AAPH was measured as oxidation of dichlorodihydrofluorescein diacetate (H(2)DCF-DA) using flow cytometry. Addition of CHL to lymphocytes in vitro significantly inhibited the increase in intracellular ROS. Further, lymphocytes from mice treated with CHL (100-400 microg/gbw i. p.) showed varying levels of ROS depending on the dose and the time (24 to 72 h) after injection. The extent of radiation-induced apoptosis and suppression of concanavalin A (con A)-induced mitogenesis ex vivo corresponded with changes in ROS levels in CHL administered mice. Antioxidant enzymes superoxide dismutase (SOD), catalase and glutathione peroxidase (GPX) were also estimated in lymphocytes from CHL-treated mice. CHL offered protection against whole body irradiation (WBI)-induced lipid peroxidation and apoptosis in lymphocytes at all the time points studied. These results demonstrate antioxidant effect of CHL in vivo. PMID- 15110093 TI - Targeted hepatic overexpression of human IRS-1: postnatal effects in the developing mouse. AB - Insulin receptor substrate-1 (IRS-1) is an intracellular docking protein involved in insulin and insulin-like growth factor (IGF) signaling. The present studies examine postnatal liver development in transgenic mice with targeted hepatic overexpression of human insulin receptor substrate-1 (hIRS-1). In mature animals, hIRS-1 overexpression augments liver growth. Based on our previous studies that have shown markedly attenuated insulin signaling in the late-gestation and early postnatal rat, we hypothesized that the liver growth effect of overexpressed hIRS 1 would be attenuated in the neonatal period. Wild-type and heterozygous transgenic mice were studied at 1, 2, 4 or 8 weeks of age. Transgene expression was seen at all ages, albeit at a lower level in the youngest animals. Liver-to carcass weight ratios were similar in hIRS-1 and wild-type mice at 1 and 2 weeks of age. At 4 and 8 weeks, transgenic mice had larger livers accounted for by increased hepatocyte number, not size. In addition, the transgenic mice had increased liver glycogen content at 8 weeks but not at 1 week. Relative to transgene mRNA expression, hIRS-1 protein levels were restricted in the younger animals. However, IRS-1-associated phosphatidylinositol-3 kinase (PI3K) activity was not similarly suppressed. Downstream from IRS-1, we found activation of the signaling kinase Akt in 8-week-old but not in 1-week-old animals. Our findings indicate that hepatic IRS-1-mediated signaling may be limited in neonatal mice at two levels, post-transcriptional down-regulation of IRS-1 content and attenuated signaling beyond the level of PI3K activation. PMID- 15110094 TI - Enzymatic description of the anhydrofructose pathway of glycogen degradation; I. Identification and purification of anhydrofructose dehydratase, ascopyrone tautomerase and alpha-1,4-glucan lyase in the fungus Anthracobia melaloma. AB - The anhydrofructose pathway describes the degradation of glycogen and starch to metabolites via 1,5-anhydro-d-fructose (1,5AnFru). The enzyme catalyzing the first reaction step of this pathway, i.e., alpha-1,4-glucan lyase (EC 4.2.1.13), has been purified, cloned and characterized from fungi and red algae in our laboratory earlier. In the present study, two 1,5AnFru metabolizing enzymes were discovered in the fungus Anthracobia melaloma for the formation of ascopyrone P (APP), a fungal secondary metabolite exhibiting antibacterial and antioxidant activity. These are 1,5AnFru dehydratase (AFDH) and ascopyrone tautomerase (APTM). AFDH catalyzed the conversion of 1,5AnFru to ascopyrone M (APM), a compound that has been earlier presumed to occur biologically, while APTM isomerized the APM formed to APP. Both enzymes were purified 400-fold by (NH(4))(2)SO(4) fractionation, hydrophobic interaction, ion-exchange and gel filtration chromatography. The purified AFDH showed a molecular mass of 98 kDa on SDS-PAGE and 230 kDa by gel filtration. The corresponding values for APTM was 60 and 140 kDa. Spectrophotometric and HPLC methods were developed for the assay of these two enzymes. To confirm that A. melaloma possessed all enzymes needed for conversion of glycogen to APP, an alpha-1,4-glucan lyase from this fungus was isolated and partially sequenced. Based on this work, a scheme of the enzymatic description of the anhydrofructose pathway in A. melaloma was proposed. PMID- 15110095 TI - CYP1A2 and NAT2 genotype/phenotype relations and urinary excretion of 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in a human dietary intervention study. AB - 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a mutagenic and carcinogenic heterocyclic amine formed during ordinary cooking, and is subsequently metabolically activated by cytochrome P4501A2 (CYP1A2) and N acetyltransferase 2 (NAT2). Respective genes encoding for these enzymes, display polymorphic distribution in the human population and are thus believed to cause interindividual differences in cancer risk susceptibility. The present study investigated the influence of dietary exposure and CYP1A2 and NAT2 genotypes and phenotypes on differential urinary PhIP excretion levels in 71 human volunteers after consumption of either a high (7.4 ng/g) or low (1.7 ng/g) dose of PhIP. Urinary PhIP excretion levels were found to reflect recent dietary exposure levels, with average levels of 174% (high dose group) and 127% (low dose group), as compared to pre-feed levels. Urinary caffeine metabolite ratios were significantly different between the two NAT2 genotypes, whereas for CYP1A2, the apparent difference in metabolic ratios between the genotypes was statistically non-significant. Significant correlations were firstly found between the CYP1A2 164A-->C (CYP1A2*1F) polymorphism and differential urinary PhIP excretion levels. Although the found correlations are driven primarily by a small number of subjects possessing the homozygous variant constellation, the strong influence of this genotype indicates that the CYP1A2*1F polymorphism could play an important role in human cancer risk susceptibility. PMID- 15110096 TI - Excretion of metabolites in urine and faeces from rats dosed with the heterocyclic amine, 2-amino-9H-pyrido[2,3-b]indole (AalphaC). AB - 2-amino-9H-pyrido[2,3-b]indole (AalphaC) is a mutagenic and carcinogenic heterocyclic amine formed during ordinary cooking. In model systems AalphaC can be formed by pyrolysing either tryptophan or proteins of animal or vegetable origin. In the present study, the in vivo metabolism of AalphaC in rats was investigated. Rats were dosed with tritium labelled AalphaC. Urine and faeces were collected over three days. The metabolites of AalphaC were characterised by HPLC-MS and quantified by liquid scintillation counting. Conjugated metabolites were characterised by enzymatic hydrolyses with beta-Glucuronidase or arylsulfatase. The data showed that the metabolic pattern of AalphaC was similar in all rats. About 55% of the dose was excreted in urine and faeces during 72 h and the major amount of AalphaC metabolites (31%) was excreted during the first 24 h. In addition to a small amount of unmetabolised AalphaC seven conjugated metabolites were characterised. Three minor metabolites were characterised as AalphaC-N(2)-glucuronide and glucuronic acid conjugates of 3-OH-AalphaC and 6-OH AalphaC. Four metabolites were all characterised as sulphuric acid conjugates and accounted for the largest amount of metabolites excreted in urine. The two major sulphuric acid conjugates were identified as AalphaC-3-O-sulfate and AalphaC-6-O sulfate, while the minor sulphuric acid conjugates were proposed to be other O sulfonated metabolites. In faeces only AalphaC was excreted and accounted for about 12% of dose during the first 24 hours. Any activated metabolites of AalphaC were not detected in rat urine or faeces. In future accumulation or binding of AalphaC to macromolecules such as DNA and proteins has to be studied. PMID- 15110097 TI - Steroid activities comparison of natural and food wrap compounds in human breast cancer cell lines. AB - In this study, we tested and compared the endocrine disruption activities of compounds in materials used to package foods (bisphenol A, bisphenol F, and bisphenol A diglycidylether BADGE) with natural molecules (genistein, apigenin, kaempferol, and tangeretin) in the human breast cancer cell lines MCF-7 (ER(+)) and MDA-MB453 (AR(+); GR(+)). Octylphenol was also chosen as a xenoestrogen reference. Two compounds had no estrogenic activity: BADGE and tangeretin. Genistein was the most active compound in the E-Screen assay with MCF-7, followed by octylphenol, bisphenol F, bisphenol A and apigenin, with kaempferol the least potent. All estrogenic compounds competed with 17beta-estradiol for binding to the MCF-7 ER and their estrogenic effects were abolished in the presence of tamoxifen, an ER antagonist. In MDA-MB453 cells transfected with pMMTVneo-Luc, all compounds had anti-androgenic activities, with octylphenol the most potent. Kaempferol, genistein, and apigenin were more potent anti-androgens than bisphenols A or F. The natural compounds had a biphasic effect on luciferase activity. At high concentrations, genistein (10(-5)M) and apigenin (10(-6)M) acted as GR agonists in transfected MDA-MB453 cells. Furthermore, apigenin, at a concentration of 10(-5)M, may act as a partial androgen receptor (AR) agonist, as nilutamide, an AR antagonist, inhibited its activity by 26%. PMID- 15110098 TI - Effect of thiabendazole on some rat hepatic xenobiotic metabolising enzymes. AB - The effect of thiabendazole (TB) on some rat hepatic xenobiotic metabolising enzymes has been investigated. Male Sprague-Dawley rats were fed control diet or diets containing 102-5188 ppm TB for 28 days. As a positive control for induction of hepatic xenobiotic metabolism, rats were also fed diets containing 1457 and 10,155 ppm butylated hydroxytoluene (BHT). Treatment with TB and BHT resulted in dose-dependent increases in relative liver weight. TB was found to be a mixed inducer of cytochrome P450 (CYP) forms in the CYP1A and CYP2B subfamilies. The administration of high doses of TB resulted in the induction of 7-ethoxyresorufin O-deethylase and 7-pentoxyresorufin O-depentylase activities, CYP1A1, CYP1A2, CYP2B1 and CYP2B1/2 mRNA levels and CYP1A2 and CYP2B1/2 apoprotein levels. In contrast, BHT was a CYP2B form inducer, increasing 7-pentoxyresorufin O depentylase activity, CYP2B1 and CYP2B1/2 mRNA levels and CYP2B1/2 apoprotein levels. Both TB and BHT induced GSH S-transferase activities towards a range of substrates. In addition, TB and BHT markedly induced GSTP1 mRNA levels, but had only a small effect on GSTT1 mRNA levels. In summary, these results demonstrate that TB induces both phase I and II xenobiotic metabolising enzymes in rat liver. PMID- 15110099 TI - Effects of extracts from Brazilian sun-mushroom (Agaricus blazei) on the NK activity and lymphoproliferative responsiveness of Ehrlich tumor-bearing mice. AB - Agaricus blazei Murrill, is an edible and medicinal mushroom which is popularly consumed due to its antitumoral properties. The immunomodulatory effects of methanol (METH), dichloromethane (DM) and n-hexane (HEX) extracts of this mushroom were evaluated in Ehrlich tumor-bearing mice. Subcutaneous inoculation of Ehrlich tumor cells inhibited the natural killer (NK) activity of spleen cells (specific lysis=6.18+/-2.56%) compared with normal mice (17.59+/-7.77%). Treatment of tumor-bearing mice with the extracts for 10 days restored the natural killer activity against Yac-1 target cells and the best results were observed by treatment with the HEX extract (21.48+/-5.26%). Treatment of the animals with the HEX extract for 10 days was also able to stimulate the mitogen induced lymphoproliferative activity of spleen cells. Thirty days after the treatment, all groups presented low proliferative activity. Specific antibody production was observed to be higher in the groups treated with the DM or METH extract 30 days after the treatment. Analysis of the 3 extracts by gas chromatography mass spectrum (GCMS) and magnetic nuclear resonance (MNR) showed that the HEX extract contains mainly sugar and fatty acids and that the METH extract also contains sugar and possibly amino acids. PMID- 15110100 TI - Effects of androstenedione on in utero development in rats. AB - This study was conducted to characterize the effect of androstenedione on estrous cyclicity, mating behavior and fetal development. Thirty-day old rats received corn oil alone or androstenedione (in corn oil) at one of four concentrations (0, 1.0, 5.0, 10.0 or 30.0 mg/kg body weight) by gavage for two weeks prior to mating, during the mating period and throughout gestation. Dose related increases in serum androstenedione, estradiol and estrone were observed in all androstenedione treated animals at gestation day 20. A statistically significant increase in serum testosterone concentration was observed in the 30 mg/kg dose group. Feed and fluid consumption were not affected by androstenedione treatment during the pre-mating or gestational periods, however a statistically significant decrease in the number of females with regular estrous cycles was observed in the 10.0 and 30.0 mg/kg dose groups. Exposure to androstenedione did not affect mean body weight gain during pre-mating or gestation. Slight not statistically significant reductions in the number of implants, number of viable fetuses and number of viable male fetuses were observed in the 30.0 mg/kg androstenedione group. Reductions were not observed in the number of corpora lutea. Fetal growth in terms of fetal weight, crown-rump length, anogenital distance and the number of external abnormalities was not affected by androstenedione exposure. At the doses given, androstenedione had no specific effect on the development of individual bones, including sternebrae. Dose related effects of androstenedione were not observed on the development of soft tissues. A statistically significant increase in moderately enlarged ureter at the kidney was observed in both the 1.0 and 5.0 mg/kg dose groups. Organ weights (expressed per gram of body weight or per gram of brain weight) were not affected by androstenedione treatment. PMID- 15110101 TI - Aluminum fluoride affects the structure and functions of cell membranes. AB - No useful biological function for aluminum has been found. To the contrary, it might play an important role in several pathologies, which could be related to its interactions with cell membranes. On the other hand, fluoride is a normal component of body fluids, soft tissues, bones and teeth. Its sodium salt is frequently added to drinking water to prevent dental caries. However, large doses cause severe pathological alterations. In view of the toxicity of Al(3+) and F(-) ions, it was thought of interest to explore the damaging effects that AlF(3) might induce in cell membranes. With this aim, it was incubated with human erythrocytes, which were examined by phase contrast and scanning electron microscopy, and molecular models of biomembranes. The latter consisted of large unilamellar vesicles (LUV) of dimyristoylphosphatidylcholine (DMPC) and bilayers of DMPC and dimyristoylphosphatidylethanolamine (DMPE) which were studied by fluorescence spectroscopy and X-ray diffraction, respectively. In order to understand the effects of AlF(3) on ion transport (principally sodium and chloride) we used the isolated toad skin to which electrophysiological measurements were applied. It was found that AlF(3) altered the shape of erythrocytes inducing the formation of echinocytes. This effect was explained by X-ray diffraction which revealed that AlF(3) perturbed the structure of DMPC, class of lipids located in the outer monolayer of the erythrocyte membrane. This result was confirmed by fluorescence spectroscopy on DMPC LUV. The biphasic (stimulatory followed by inhibitory) effects on the isolated skin suggested changes in apical Cl(-) secretion and moderate ATPase inactivation. PMID- 15110102 TI - Safety evaluation of phosphodiesterase derived from Leptographium procerum. AB - 5'-Phosphodiesterase is produced by fermentation of the fungus Leptographium procerum and is used to hydrolyse yeast RNA to produce flavour enhancers. To establish the safety in use of this enzyme preparation a number of studies have been performed: analysis for the potential of the production strain to produce toxic secondary metabolites, 28-days oral toxicity study of the preparation in the rat, bacterial mutation assay and in vitro mammalian chromosome aberration test in human lymphocytes. The production strain did not produce any secondary metabolites that may be of significance in food. Administration of dosage levels of 1250, 2500 and 5000 mg/kg body weight/day to rats for 28 day did not result in any toxicological significant changes. The enzyme preparation showed no mutagenic activity in the bacterial mutation assay and no clastogenic potency in an in vitro test. These results together with existing knowledge of the production organism and the chemical and microbiological characterisation of the enzyme preparation lead to the conclusion that the enzyme preparation containing 5' phosphodiesterase activity from Leptographium procerum can safely be used for the production of flavour enhancers from bakers yeast at the anticipated intake levels for these uses. PMID- 15110103 TI - Reproduction of the tundra vole (Microtus oeconomus) with dietary phytosterol supplement. AB - Phytosterols (PS) are plant-derived compounds with estrogenic activity in vitro and beneficial effects on the serum lipid profile in vivo. In nature, PS exposure can derive from pulp mill effluents. The effects of a pulp-mill derived PS mixture on the reproduction, endocrine variables and enzyme activities of the tundra vole (Microtus oeconomus) were investigated in a two-generation study. The cumulative food intake of PS-treated females was higher than in the control group supporting previous results on the effects of PS on food consumption in rodents. 85% of the PS treated pairs reproduced, but the figure was only 60% for the control pairs. The plasma and testicular testosterone concentrations were lower in the adult PS males, but the PS-treated male offspring had higher testicular testosterone concentrations than their controls. In the female offspring, the liver lipase activity was higher in the PS-treated group, which could be a result of decreased cholesterol absorption in the gut. Chronic PS treatment increased the reproduction probability of the species and had a potential effect on the sex steroid hormones of maturing offspring, which could have applications in environmental monitoring. PMID- 15110104 TI - Induction and recovery of hepatic drug metabolizing enzymes in rats treated with Ginkgo biloba extract. AB - Herb-drug interactions, especially cytochrome P450 (CYP)-mediated interactions, cause an enhancement or attenuation in efficacy of co-administered drugs. In a previous study, we reported that repeated oral ingestion of Ginkgo biloba extract (GBE) markedly induced hepatic drug metabolizing enzymes in rats. In this study, we focused on the recovery of GBE-induced hepatic drug metabolizing enzymes after the discontinuation of GBE in rats. Feeding of a 0.5% GBE diet to rats for 1 week markedly increased liver weight, content of total CYP, activities of 6 CYP subtypes and glutathione S-transferase (GST). The content and activities of CYP enzymes were recovered to almost basal levels within 1 week after the discontinuation of GBE, while the activity of GST gradually decreased and recovered to the control level after 3 weeks. These results indicated that GBE induced hepatic drug metabolizing enzymes in rats, especially CYPs, were rapidly recovered by discontinuation of GBE in rats even after excess treatment, and suggested that interactions of GBE with drugs could be avoided by discontinuation of GBE. PMID- 15110105 TI - The toxicology and safety of apple polyphenol extract. AB - Apple polyphenol extract has strong antioxidant activity and various physiological functions, and is used in Japan as a food additive and nutritional supplements. Here, we tested the consumption safety of Applephenon, which is a polyphenol extract produced from unripe apples. The Ames test without S9 mixture revealed that Applephenon, had slight mutagenicity at a high concentration of 2500 microg/plate; however, both chromosomal aberration test and the micronucleus test found no significant mutagenicity. Furthermore, an acute oral-toxicity test, and a 90-day subchronic-toxicity test showed no significant hematological, clinical, chemical, histopathological, or urinary effects at a dose of 2000 mg/kg. These results confirm that Applephenon is safe and no toxic at average dietary level. PMID- 15110106 TI - Genotoxic evaluation of a vinifera skin extract that present pharmacological activities. AB - Toxicity of an alcohol-free hydro-alcoholic grape skin extract (GSE) obtained from red grapes Vitis labrusca (Isabel varietal) that present antihypertensive, vasodilator and antioxidant effects was estimated by different bioassays. Using the Salmonella/microsome assay for strains TA97, TA98, TA100 and TA102 no mutagenicity was detected for all tested concentrations (0.1-100 microg/ml), even with metabolization. Nevertheless, cytotoxicity was observed for TA97 and TA102 with and without metabolization and for TA100 with metabolization. The measurement of beta-galactosidase induction in the SOS-chromotest was positive only for Escherichia coli PQ37 when metabolization enzymes were present. Using Balb/c 3T3 fibroblasts, DNA strand breaks induction by GSE was also investigated by the comet assay and no significative difference was detected for treated and no treated DNA for 60 min. Our data suggest that GSE although no mutagenic presents cytotoxic activity. PMID- 15110107 TI - Inhibitory effects of Luobuma tea and its components against glucose-mediated protein damage. AB - Luobuma tea, prepared from the leaves of Apocynum venetum L., is a popular beverage in China. In this study, the activity of Luobuma leaf extract and its components against the formation of advanced glycation endproducts (AGEs), which are largely involved in the pathogenesis of diabetic vascular complications, was examined using the in vitro glycation reaction. Strong inhibitory activity against the formation of AGEs was shown by Luobuma aqueous extract. Following further fractionation of this extract, seven polyphenolic compounds, i.e. (+/-) gallocatechin, (-)-epigallocatechin, (+/-)-catechin, (-)-epicatechin, epicatechin (4beta-8)-gallocatechin, epigallocatechin-(4beta-8)-epicatechin and procyanidin B 2, were isolated by Sephadex LH-20 column chromatography. These purified compounds also exerted inhibitory activities that were more potent than the positive control, aminoguanidine. Our findings may help to explain the beneficial effects of this plant against atherosclerosis. PMID- 15110108 TI - Study on anti-androgenic effects of bisphenol a diglycidyl ether (BADGE), bisphenol F diglycidyl ether (BFDGE) and their derivatives using cells stably transfected with human androgen receptor, AR-EcoScreen. AB - We studied in vitro hormonal activity of bisphenol A diglycidyl ether (BADGE) and bisphenol F diglycidyl ether (BFDGE), which are used as a material of interior coating for food cans. We also examined related compounds such as 2,2-bis[4-(3 chloro-2-hydroxypropoxy)phenyl]propane (BADGE.2HCl), and bis[4-(3-chloro-2 hydroxypropoxy)phenyl]methane (BFDGE.2HCl) etc. For this purpose, we constructed two stably transfected CHO-K1 cell lines (AR-EcoScreen for androgenic activity and c-luc for cell toxicity evaluation). One stably expresses luciferase with induction of androgen. The other stably expresses luciferase without androgen induction. Also, we have determined the androgenic and anti-androgenic effects of the test chemicals by reporter gene assay with these cell lines. None of the chemicals tested by this assay exhibited androgen agonistic activity. However, BADGE.2HCl and BFDGE.2HCl had the conspicuous antagonistic activity for androgen. These compounds had a high binding affinity for androgen receptor. Furthermore, these two compounds did not show the estrogenic activity in vitro assays. On the contrary, bisphenol A and bisphenol F exhibited anti-androgenic activity in vitro in addition to the estrogenic activity. These results suggest that these chlorohydroxy compounds of BADGE and BFDGE act as androgen antagonist through the process of binding to androgen receptor. PMID- 15110109 TI - St. John's Wort (Hypericum perforatum) induces overexpression of multidrug resistance protein 2 (MRP2) in rats: a 30-day ingestion study. AB - St. John's Wort (Hypericum perforatum, SJW) has been used as a herbal medicine for the treatment of depression in oral doses of 900-1050 mg/day in humans. However, the ingestion of SJW was reported to cause interactions with drugs. In the present study, we examined the effects of SJW treatment on the induction of drug transporters and enzymes in rats. An immunoblot analysis was performed to quantify the expression of the transporters and enzymes. SJW was given at a dose of 400 mg/kg/day, since it was reported that 400 mg/kg/day is antidepressant effective dose in rats. When SJW was administered for 10 days, the amounts of multidrug resistance protein 2 (MRP2), glutathione S-transferase-P (GST-P) and cytochrome P450 1A2 (CYP1A2) in the liver were increased to 304%, 252% and 357% of controls, respectively, although the amounts of P-glycoprotein and multidrug resistance protein 1 were not changed. Under the same conditions, an increase of MRP2 in the kidney was not observed. The increase in the levels of each protein was maximal at 10 days after SJW treatment and lasted for at least 30 consecutive days. These results suggest that SJW induces hepatic MRP2, GST-P and CYP1A2 overexpressions, and thus, it could affect drug metabolism, conjugation and disposition. PMID- 15110110 TI - Results of a 13 week safety assurance study with rats fed grain from glyphosate tolerant corn. AB - The current study presents the results of a 13 week feeding study in rats with grain from Roundup Ready corn which is tolerant to the herbicide glyphosate. Herbicide tolerance was accomplished through the introduction of cp4 epsps coding sequences into the corn genome for in planta production of CP4 EPSPS enzymes. Unlike related corn EPSPS enzymes, CP4 EPSPS enzymes are not inhibited by the herbicide glyphosate. Purina TestDiets formulated Roundup Ready corn grain into rodent diets at levels of 11 and 33% (w/w). The responses of rats fed diets containing Roundup Ready corn grain were compared to that of rats fed diets containing non-transgenic grain (controls). All diets were nutritionally balanced and conformed to Purina Mills, Inc. specifications for Certified LabDiet 5002. There were 400 rats in the study divided into 10 groups of 20 rats/sex/group. Overall health, body weight, food consumption, clinical pathology parameters (hematology, blood chemistry, urinalysis), organ weights, gross and microscopic appearance of tissues were comparable between groups fed diets containing Roundup Ready and control corn grain. This study complements extensive agronomic, compositional and farm animal feeding studies with Roundup Ready corn grain, confirming it is as safe and nutritious as existing commercial corn hybrids. PMID- 15110111 TI - Toxicity of trans-nonachlor to Sprague-Dawley rats in a 90-day feeding study. AB - The chlordane constituent trans-nonachlor and its metabolite oxychlordane are among the most persistent chlordane-related contaminants and are found in tissues and milk from humans ingesting diets high in Arctic marine mammal fat. Although chlordane is no longer registered in North America, there is a need for toxicological data on chlordane-related contaminants found in food and the environment which are either structurally different or relatively more abundant than the constituents of the original chlordane mixture. Thus, a feeding study was undertaken to provide toxicological data on trans-nonachlor. Male and female Sprague-Dawley rats were exposed to 0, 5, 13 or 50 ppm trans-nonachlor in feed for 90 days and clinical, hematological and histopathological changes were assessed in each rat. Female rats were less able than males to metabolize and eliminate trans-nonachlor and, as a result, accumulated more trans-nonachlor in their adipose tissues. trans-Nonachlor, like technical chlordane and other organochlorines, induced liver microsomal enzymes in a pattern similar to phenobarbital. Endocrine effects included functional and morphological changes in the thyroid and adrenals. In male rats exposure to trans-nonachlor was associated with changes in endpoints indicative of increased oxidative stress, which may be related to both direct action on cellular targets or to secondary effects resulting from cytochrome P450 induction. The results indicate that subchronic trans-nonachlor exposure in rats induced hepatic changes with far-reaching metabolic and endocrine effects. Differences in target organ responses in male and female rats indicate that the sex-related metabolic differences affecting trans-nonachlor bioaccumulation and elimination merit further study. PMID- 15110112 TI - Determining the safety of chromium tripicolinate for addition to foods as a nutrient supplement. AB - Trivalent chromium is an essential element required for normal carbohydrate, lipid, and protein metabolism in humans and animals. This article describes how one particular form of trivalent chromium, chromium tripicolinate (Chromax), a stable complex of trivalent chromium and picolinic acid, was determined to be safe for use as a nutrient supplement in foods (i.e., nutritional bars and beverages). A safety assessment of a nutrient supplement for use in food requires an evaluation of the safety of the product (Chromax), as well as an evaluation of the safety of the intended dietary ingredient (chromium tripicolinate). In this assessment, the production process, final product specifications, and product analysis results for Chromax were evaluated to determine the safety of the final product. Then, the Estimated Daily Intakes (EDIs) of trivalent chromium and picolinic acid from the proposed use of Chromax were calculated using data from the USDA's most recent Continuing Survey of Food Intakes by Individuals. The EDI for trivalent chromium was compared to the Acceptable Daily Intake (ADI) for this compound derived from animal studies, corroborated with human data, while the EDI for picolinic acid was compared to existing in vivo exposure to this compound that occurs naturally in the body. As long as the EDI of trivalent chromium is less than the ADI for this compound and the EDI of picolinic acid is less than its estimated in vivo exposure, Chromax can be considered safe for its intended use. Employing this approach, it was determined that Chromax is safe, as well as generally recognized as safe (GRAS), for use in nutritional bars and beverages as a nutrient supplement at a maximum use level of 2.4 mg of Chromax per product serving. PMID- 15110114 TI - The six golden rules to improve compliance in hand hygiene. AB - Improvement of compliance in hand hygiene is probably the most effective step in reducing the incidence of nosocomial infections (NI). But improvement of compliance is known to be complex. Six possibilities for improving compliance are available although some of them may be difficult to carry out. Rule 1: Select an alcohol-based hand rub which has a good skin tolerance and is acceptable to health care workers to use. This has been shown to improve compliance. Rule 2: The hand rub shall be easily available. Wall dispensers near the patient and pocket bottles may well help. Other possibilities should be assessed locally. Rule 3: Implement teaching and promotion of hand hygiene, which has been shown to be very effective. This is may be the most effective tool but will cost time and money. If money is a problem, rule 4 may be the solution. Rule 4: Create a hospital budget which covers all costs involved with preventable nosocomial infection. Combine it with the budget for hand hygiene products. Even a small number of prevented NI largely outweighs the cost of effective hand hygiene products. Rule 5: Get senior staff to set a good example in order to motivate junior staff, because negligence in hand hygiene appears to correlate with the number of professional years. Rule 6: Have the patient-staff ratio well balanced. It has been shown that staff shortage decreases hand hygiene compliance. Other factors may be important as well, but implementation of these 6 golden rules could be an effective step into the right direction. PMID- 15110115 TI - European norms in hand hygiene. AB - In the area of hand hygiene, European Norms exist, or are under development, with regard to protective gloves and for assessing the antimicrobial efficacy of hand disinfectants. Important norms for gloves are EN 420 (General requirements), EN 374 (Protective gloves against chemicals and microorganisms) and EN 455 (Medical gloves for single use). A suspension test for the demonstration of bactericidal activity (prEN 12054) is obligatory for hand disinfectants in all fields of application; a test to prove activity against yeasts applies only to hygienic hand rub. (Optional) Claims for virucidal activities can be substantiated by prEN 1476 and, in future, for mycobactericidal capacity by a test which is still under development. In vivo tests exist for post-contamination treatments, hygienic hand wash and hygienic hand rub (EN 1499 and EN 1500 respectively), and for the preoperative surgical hand rub/wash (prEN 12791). The two former tests employ artificially contaminated hands, the latter test is done with clean hands. All in vivo tests use reference hand treatments (with unmedicated soap or 2-propanol 60% (vol.) or 1-propanol 60% (vol.), respectively) against the results of which are compared with those achieved with the product under test and with the same volunteers. An antiseptic soap needs to be significantly more efficacious than unmedicated soap, a product for hygienic hand rub must not be inferior to the reference treatment with 2-propanol, and a surgical hand disinfectant must not cause a smaller bacterial reduction than the reference preparation with 1 propanol, immediately, and after 3 h. An (optional) claim for sustained activity of a surgical disinfectant needs to be demonstrated by achieving a significantly stronger bacterial reduction after 3 h than the reference preparation. PMID- 15110116 TI - Provision of alcohol hand rub at the hospital bedside: a case study. AB - A pilot study was performed on a 28-bed acute hospital ward, promoting hand disinfection by providing Sterillium alcohol hand rub at the bedside. Bottles of Sterillium with pump dispensers were attached to the end of each patient's bed, next to the observation charts. Posters promoting the use of the product were prominently displayed and several ward staff attended a presentation on the effectiveness of Sterillium. Although this was a small study, Alert Organism rates during the three-month trial indicated a reduction in methicillin-resistant Staphylococcus aureus (MRSA) incidence and an increased Clostridium difficile incidence. Several other factors that may have influenced the study outcomes will be discussed. There were no health and safety problems during the trial, but other issues were identified which have implications for introducing the bedside rub on a hospital wide basis, including the type of holder used, logistical arrangements and the staff and patient information requirements. The strategies adopted to address these issues will be discussed. PMID- 15110117 TI - Efficacy of alcohol-based gels compared with simple hand wash and hygienic hand disinfection. AB - A recent research letter on the limited efficacy of alcohol-based hand gels has alerted the global infection control community and raised the question of the true significance of data obtained according to EN 1500. It has been described that a 1 min simple hand wash reduces artificial contamination of hands by a log(10) reduction factor of 2.8 and a 1 min reference hand disinfection with 2 propanol (60%, v/v) by a factor of 4.6 steps. The EN 1500 gel data show that the 30 s efficacy of most gels is closer to a simple hand wash than to the reference hand disinfection. The 30 s efficacy of most alcohol-based liquid products and one gel, however, is almost identical to the reference hand disinfection. In many European countries alcohol-based liquid products have been established as a standard practice in hygienic hand disinfection for decades. Replacement of these products with most available gels would be a step backward in terms of efficacy and has still to be seen critically from the efficacy point of view. PMID- 15110118 TI - The importance of cleaning for the overall results of processing endoscopes. AB - Reprocessing comprises three steps: cleaning, disinfection and-if required sterilisation. While the extents of disinfection and of sterilisation are quantitatively defined, there are only imprecise (qualitative) definitions of cleaning. There are two main reasons for accurate cleaning. First organic and inorganic materials that remain on inner and outer surfaces will interfere with the efficacy of the disinfectants. In case of endoscopes this will lead to channel blockages; they remain undisinfected. Second the bioburden found on endoscopes after use can be very high. Data available demonstrate that a bacterial burden of up to 10(9)cfu/endoscope channel can be expected. Therefore it is necessary to perform a thorough cleaning. Studies using endoscopes showed a reduction in microbial counts by a factor of approximately 10(4) by cleaning (manual and mechanical). Therefore in 2001 the German Society of Hospital Hygiene (DGKH) specified its requirements and recommendations for determining cleaning efficacy separately from those for disinfection. Cleaning and disinfecting can be done manually or mechanically, but it seems impossible to validate manual processes. However our studies in two different washer-disinfectors (WD) showed differences in cleaning efficacy. The tested cleaning processes showed different efficacies. Not all cleaning processes showed better results than water alone with regard to visible cleanliness and to a microbiological reduction E. faecium. Our results show that the evaluation of cleanliness exclusively by visible inspection is not sufficient, particular for the lumens of endoscopes. The results also show that a cleaning process may be very effective also in reducing micro-organisms present. PMID- 15110119 TI - Reprocessing endoscopes: national recommendations with a special emphasis on cleaning--the German perspective. AB - Transmission of pathogens through endoscopes or endoscopic procedures has been documented frequently, although reliable data on the incidence of endoscope associated infections do not exist. In order to control such infections as well as to ensure adequate quality management in endoscopy, the German Robert Koch Institute has produced recommendations for hygienic processing of flexible endoscopes and accessory devices. Reprocessing using a washer disinfector is explicitly preferred to manual procedures. Control measures are focused on microbiological sampling and limiting values have been defined for rinsing fluids taken from the channels of the scopes. Although the guideline leaves a number of questions open, e.g. test methods to control the efficacy of the cleaning process. It helps to establish quality management in the field of endoscopy and to improve the level of safety both for patients and staff. PMID- 15110120 TI - Reprocessing endoscopes: United States perspective. AB - Endoscopes are used frequently for the diagnosis and therapy of medical disorders. For example, greater than 10000000 gastrointestinal endoscopic procedures are performed each year in the United States. Failure to employ appropriate cleaning and disinfection/sterilization of endoscopes has been responsible for multiple nosocomial outbreaks and serious, sometimes life threatening, infections. Flexible endoscopes, by virtue of the site of use, have a high bioburden of microorganisms after use. The bioburden found on flexible gastrointestinal endoscopes following use has ranged from 10(5) to 10(10)CFU/ml, with the highest levels being found in the suction channels. Cleaning dramatically reduces the bioburden on endoscopes. Several investigators have shown a mean log(10) reduction factor of 4 (99.99%) in the microbial contaminants with cleaning alone. Cleaning should be done promptly following each use of an endoscope to prevent drying of secretions, allow removal of organic material, and decrease the number of microbial pathogens. Because the endoscope comes into intimate contact with mucous membranes, high-level disinfection is the reprocessing standard after each patient use. High-level disinfection refers to the use of a disinfectant (e.g., FDA-cleared chemical sterilant or high-level disinfectant) that inactivates all microorganisms (i.e., bacteria, viruses, fungi, mycobacteria) but not high levels of bacterial spores. The disinfection process requires immersion of the endoscope in the high-level disinfectant and ensuring all channels are perfused for the approved contact time (e.g., for ortho phthaladehyde this is 12 min in the US). Following disinfection, the endoscope and channels are rinsed with sterile water, filtered water, or tapwater. The channels are then flushed with alcohol and dried using forced air. The endoscope should be stored in a manner that prevents recontamination. A protocol that describes the meticulous manual cleaning process, the appropriate training and evaluation of the reprocessing personnel, and a quality assurance program for endoscopes should be adopted and enforced by each unit performing endoscopic reprocessing. PMID- 15110121 TI - Importance of cleaning for reprocessing endoscopes and thermolabile sterile medical devices: French use and regulations. AB - In France, endoscope maintenance regulations present some particularities in terms of the definitions and texts referring to the disinfection and sterilization of medical devices, with respect to the prion risk. The main measures specified are a double cleaning prior to disinfection, with some stipulations concerning the procedure itself, such as time limits, duration and rinsing. The use of aldehydes in cleaning products is prohibited and it is recommended that peracetic acid, or any chlorinated product, be used at the disinfection phase. For machines, the recycling of detergents or disinfectants is prohibited, and traceability procedures are mandatory. The French Agency for Safety of Health Products (AFSSAPS) is committed to providing standards that prevent any undesirable consequences for the patient, the operator or the equipment. All these measures will be described in a 'user's guide' intended for medical care units, to be released by the National Technical Committee on Nosocomial Infections (CTIN). PMID- 15110122 TI - Test models to determine cleaning efficacy with different types of bioburden and its clinical correlation. AB - The importance of cleaning as a first crucial step in reprocessing instruments and endoscopes is recognized worldwide. However, no standards to determine the efficacy of cleaning have been established. We have therefore investigated Bodedex forte, a new cleaner, in various test models derived from critical types of bioburden on flexible endoscopes. Removal of dried blood from metal carriers was determined in comparison with standard instrument disinfectants. Removal of biofilm endotoxin from silicone test pieces and removal of dried X-ray contrast medium from polyethylene pieces was measured in comparison with one other standard cleaner. Residual bacteria in a biopsy channel from duodenoscopes following use of Bodedex forte, compared with two other cleaners, were measured in an endoscopy unit. After 15 min exposure to Bodedex forte, 95% of the dried blood were removed. Removal was between 0 and 86% with the disinfectants. Bodedex forte reduced endotoxin by 1.91+0.19 log(10)-steps compared with 0.43+0.19 log(10)-steps Cidezyme (P < 0.001) two-sided t-test). Removal of dried X-ray contrast medium was 99% with Bodedex forte and 94% with a conventional cleaner. No bacterial contamination after reprocessing was found in 98% of duodenoscopes with Bodedex forte (78 duodenoscopes), in 72% with a conventional cleaner (129 duodenoscopes) and in 69% with an enzymatic cleaner (100 duodenoscopes). The difference between the three cleaners was significant (P < 0.001) chi-squared test). The superiority of the cleaning capacity of the new cleaner was demonstrated in various test models, which were designed according to the clinical relevance of different bioburdens. Implementation of accepted and reproducible standards for testing the cleaning efficacy will remain a goal for the next years. PMID- 15110123 TI - Surrogate viruses for testing virucidal efficacy of chemical disinfectants. AB - Since important agents of viral nosocomial infections like hepatitis B and C viruses and norovirus do not replicate sufficiently in cell culture systems, disinfectants with suspected efficacy against these viruses must be evaluated by different methods. Besides molecular approaches and indirect tests, the use of surrogate viruses with similar biophysical properties and genomic structure allows the assessment of virucidal efficacy of chemical disinfectants in quantitative suspension tests. Furthermore, insights into the survival of these viruses in the environment are possible. In recent years, duck hepatitis B virus and bovine viral diarrhoea virus have been tested as surrogates for hepatitis B and C viruses. Feline calicivirus serves as a surrogate for the group of norovirus. By including these viruses in inactivation experiments, valuable data from suspension tests can be derived on the virucidal efficacy of chemical disinfectants. Even in vivo tests using fingerpads of adult volunteers can be performed with these animal viruses without risk of infection. In contrast to in vitro examinations, the results of these tests allow use recommendations of chemical disinfectants for outbreak situations and daily routine disinfection. PMID- 15110124 TI - Nosocomial infection surveillance and control activities in Spain under HELICS and NosoMed programs frame. AB - Since the 1980s, a number of multi-center studies for the surveillance of nosocomial infections have been developed in Spain. The first of them, EPINE (study on the prevalence of nosocomial infections in Spain), was followed by two incidence-based surveillance systems: ENVIN-UCI, for intensive care units, and PREVINE, applicable to a broader range of hospital areas. These surveillance systems have made it possible for Spain to participate in two European projects, namely HELICS and NosoMed. It is expected that the European countries participating in these projects will have substantial returns in terms of better prevention and control of health care-related infections. PMID- 15110125 TI - Assessment of new chemical disinfectants for HBV virucidal activity in a cell culture model. AB - Several new chemical disinfectants were processed for Hepatitis B virus (HBV) virucidal activity in a cell culture model. A pooled HBV infected human plasma with 10(10.4) HBV DNA copies/mL was treated with the tested disinfectant. It was then subjected, for three days at several dilutions, to cell culture using the human hepatoma cell line, HepG2, with 4% polyethyleneglycol and 3 mM sodium butyrate. Thirty-seven assays were performed on 12 products, with up to 3 concentrations and 3 time exposures for each product tested. The mean viral titre without disinfectant was 10(5.18) infectious units per mL. Our results showed that products all four hand rubs examined, two of the three surface disinfectants and two of the three instrument disinfectants were highly active whatever concentrations and time exposures, reducing viral times by factors of 10(3) 10(4). However, other products such as one of the surface disinfectants was only active at concentrations above 0.5% for 15 min. Similarly the skin disinfectant, one of the instrument disinfectants and the hand wash agent (diluted to 50%) were less or not active (of <10(3) fold reduction). This is the first study using a cell culture model to assess virucidal activity against HBV of new disinfectants. It showed that most 9/12 products were active by either HBs antigen alteration (8/9) or probable envelope disruption (1/9). Further studies are in progress using this model to assess the activity of other chemical disinfectants such as peracetic acid against HBV. PMID- 15110126 TI - Microbicides and the environmental control of nosocomial viral infections. AB - Viruses are important causes of acute and chronic diseases in humans. Newer viruses are still being discovered and those that are already known are being incriminated in the aetiology of clinical conditions with hitherto unknown causes. Apart from frequently causing infections in the general community, many types of viruses are also significant nosocomial pathogens. While it is generally agreed that we underestimate the proportion of nosocomial infections that are viral, due to a lack of routine monitoring, viruses easily account for more than 30% of the cases of hospital-acquired infections in many paediatric settings. Indeed, the relative importance of viruses in this respect is increasing due to a number of societal and demographic changes as well as alterations in healthcare practices. Safe vaccines against many common nosocomial viral agents are currently unavailable while there is also a virtual lack of effective and affordable chemotherapy against them. There is, therefore, renewed emphasis on preventive strategies by better understanding of the relative importance of various vehicles in the nosocomial spread of viruses and by infection control using microbicides. This, in turn, has stimulated considerable interest in the development of formulations that are not only safer but which also have demonstrated activity against major types of nosocomial viral pathogens. Further, much work is now underway to design better methods to assess the virucidal activity of microbicides used to decontaminate hands, reusable medical devices and environmental surfaces in critical areas of healthcare settings. It is anticipated that these approaches will result in reducing the health and economic impact of nosocomial infections due to viruses. PMID- 15110127 TI - Household cleaning and surface disinfection: new insights and strategies. AB - Recently, new insights into the persistence of pathogens, their transfer from inanimate surfaces to humans and the risk of contamination and dissemination of pathogens by detergents have been gained. Furthermore, new experimental data on the interruption of chains of infection by disinfectants as well as results of outbreak-control studies are now available. Hence it has become necessary to reassess the potential benefits using disinfectants to prevent and control nosocomial infections. Based on the new findings and in view of the increasing incidence of nosocomial infections and antibiotic resistances, the German Robert Koch-Institut has issued completely revised recommendations on Household Cleaning and Surface Disinfection. With respect to these recommendations we developed a new test method, which allows comparison of the efficacy of disinfection in reducing the microbial loads and their dissemination with that of cleaning procedures under practical conditions. In a multi-factor approach, mechanical properties (wet mop technique), utensils (different mop materials) and active agents (disinfectant, detergent) were taken into consideration. We found that under the given conditions, dissemination of the test organism Staphylococcus aureus did not take place when using aldehydes and peroxides, it did take place, however, when water, surfactants, and the disinfectants glycol derivatives, quaternary ammonium compounds and alkylamines were used. PMID- 15110128 TI - Association of dyslipidemia and effects of statins on nonmacrovascular diseases. AB - BACKGROUND: Statins have mechanisms of action that expand their effects beyond cholesterol lowering and atherosclerotic medical conditions. OBJECTIVE: This review summarizes clinical evidence for the association of dyslipidemia and the effects of statin use on aortic stenosis, Alzheimer's dementia (AD), osteoporosis, prevention of diabetes mellitus (DM), diabetic retinopathy, age related macular degeneration, and diabetic/nondiabetic nephropathy. METHODS: An English-language literature search was conducted using MEDLINE (1966-June 2003). Bibliographies of retrieved articles were reviewed. Search terms included statin, HMG-CoA reductase inhibitors, aortic stenosis, Alzheimer's dementia, osteoporosis, prevention of diabetis, diabetic retinopathy, age-related macular degeneration, diabetic nephropathy, and nondiabetic nephropathy. RESULTS: Three retrospective cohort trials have shown an association between statin use and the progression of aortic stenosis; one of these trials observed a 45% decrease in aortic valve area in 1 year. In AD, one cross-sectional analysis found 60% to 73% lower AD rates in lovastatin or pravastatin recipients ( P<0.001 ). Of the multiple observational studies on the effect of statins on fracture risk, some have shown a decreased risk, with an odds ratio as low as 0.50 (95% CI, 0.33 0.76); others have demonstrated no association. A post hoc analysis of the West of Scotland Coronary Prevention Study found a 30% reduction in the development of DM ( P=0.042 ), but this was not duplicated in the Anglo-Scandinavian Cardiac Outcomes Trial-Lipid Lowering Arm. A small clinical trial of 6 patients (11 eyes) demonstrated improved retinal hard exudates with pravastatin treatment in patients with diabetic retinopathy. In a cross-sectional analysis, age-related macular degeneration was found to be less common among statin users than nonusers (4% [ 1/27 ] vs 22% [ 76/352 ]; P=0.02. Multiple small clinical trials of 19 to 56 patients with diabetic and nondiabetic nephropathy at various stages generated inconsistent results for an association between statin use and decreased albumin excretion rate and decreased rate of decline in glomerular filtration. CONCLUSION: Data of variable quantity and quality support the use of statins as adjuncts in the treatment of nonmacrovascular diseases. PMID- 15110129 TI - Enfuvirtide: a fusion inhibitor for the treatment of HIV infection. AB - BACKGROUND: Drug resistance continues to be a major challenge in the treatment of HIV-1 infection. Virtually all currently available antiretroviral medications inhibit the viral reverse transcriptase or protease. Enfuvirtide is the first fusion inhibitor approved by the US Food and Drug Administration for use in combination with other antiretroviral agents for the treatment of HIV-1 infection in treatment-experienced patients. OBJECTIVE: This paper describes the pharmacologic properties and clinical usefulness of enfuvirtide. METHODS: Relevant information was identified through searches of MEDLINE (1990 to October 2003), International Pharmaceutical Abstracts (1970 to October 2003), and meeting abstracts of major HIV/AIDS conferences (1996-2003) using the search terms enfuvirtide, pentafuside, T-20, DP-178, and fusion inhibitor. RESULTS: In vitro, enfuvirtide exhibits activity against HIV-1 isolates that are resistant to all other classes of anti-retroviral medications. Enfuvirtide blocks the entry of HIV 1 into host cells by interfering with virus-cell fusion, making it unique among licensed antiretroviral medications. In human adults, enfuvirtide has a volume of distribution of 5.48 L, is highly bound to plasma protein (92%), has a plasma elimination half-life of 3.8 hours, and is catabolized by peptidases and proteinases in various tissues. Dose adjustment does not appear necessary on the basis of age, race, or body weight, but may be warranted in women weighing <50 kg. A literature review did not identify any data on the disposition of enfuvirtide in patients with hepatic or renal insufficiency. Clinical trials suggest that enfuvirtide reduces plasma HIV-1 RNA levels in highly treatment experienced patients taking an optimized antiretroviral regimen. Pivotal trials indicated a mean change in HIV-1 RNA of -1.48 log(10) copies/mL in the enfuvirtide arm at week 48, compared with -0.63 log(10) copy/mL in the control arm ( P<0.001 ). The mean absolute increase on CD4 cell count was 46 cells/mm(3) (91 cells/mm(3)) in the enfuvirtide arm vs 45 cells/mm(3) in the control arm; P<0.001 ). The most commonly reported (>15 cases per 100 patient-years of exposure) adverse events (AEs) in clinical trials included injection-site reactions, diarrhea, nausea, fatigue, insomnia, peripheral neuropathy, headache, vomiting, and fever. The most commonly reported (> or =2%) laboratory abnormalities (grade III or IV) were eosinophilia, anemia, and increases in amylase, lipase, triglycerides, creatine phosphokinase, alanine aminotransferase, aspartate aminotransferase, and gamma-glutamyl transferase. In clinical trials, serious AEs leading to study discontinuation occurred in 12.9% ( 114/885 ) of patients in the enfuvirtide arm, compared with 10.7% ( 12/112 ) in the control arm ( P = NS ). The recommended dosage of enfuvirtide is 90 mg SC BID in adults and 2 mg/kg SC BID in children. Efficacy studies in children are ongoing. CONCLUSION: Although additional studies are needed, enfuvirtide appears to be a promising agent, in combination with other antiretroviral agents, for the treatment of HIV infection in treatment-experienced patients. PMID- 15110130 TI - Effects of simvastatin on the lipid profile and attainment of low-density lipoprotein cholesterol goals when added to thiazolidinedione therapy in patients with type 2 diabetes mellitus: A multicenter, randomized, double-blind, placebo controlled trial. AB - BACKGROUND: Coronary heart disease is the major cause of mortality in individuals with diabetes mellitus (DM). Given the increasingly aggressive low-density lipoprotein cholesterol (LDL-C) goals for patients with DM set by the National Cholesterol Education Program Adult Treatment Panel III and the American Diabetes Association, many patients remain above target. Treatment with thiazolidinediones (TZDs) improves glycemic control but does not lower (and may raise) LDL-C concentrations. OBJECTIVE: This study assessed the lipid-modifying efficacy and tolerability of adding the hydroxymethylglutaryl coenzyme A-reductase inhibitor simvastatin to existing TZD therapy in patients with type 2 DM. METHODS: This was a multicenter, randomized, double-blind, placebo-controlled, parallel-group trial. Patients with type 2 DM who were taking a stable dose of pioglitazone or rosiglitazone and had a glycosylated hemoglobin (HbA1c) value < or =9.0% and an LDL-C concentration > 100 mg/dL were randomized to receive simvastatin 40 mg (the recommended initial dose for patients with DM) or placebo for 24 weeks. The primary end point was the effect of treatment on LDL-C concentrations. Other lipid, lipoprotein, and safety measures were also assessed. RESULTS: Two hundred fifty-three patients (127 [50.2%] men, 126 [49.8%] women; mean age, 56 years) were randomized to treatment (123 simvastatin, 130 placebo). At the end of the study, mean LDL-C concentrations were reduced 34.)% from baseline (from 134.3 to 89.5 mg/dL) in the simvastatin group and were unchanged in the placebo group (P<0.001). Simvastatin produced significant reductions in concentrations of total cholesterol, triglycerides (TG), non-high-density lipoprotein cholesterol, and apolipoprotein (apo) B compared with placebo (all, P<0.001 ) and significant increases in concentrations of high-density lipoprotein cholesterol (HDL-C) ( P=0.002 ) and apo A-I ( P=0.006 ). In patients who had not attained target concentrations of LDL-C (<100 mg/dL), TG (<150 mg/dL), or HDL-C (>45 mg/dL) at baseline, significantly more simvastatin recipients had achieved these goals at the end of the study compared with placebo recipients (LDL-C: 67.3% vs 5.2%, respectively, P<0.001; HDL-C: 95.3% vs 83.6%, P<0.05; TG: 40.8% vs 11.0%, P<0.001 ). Simvastatin was well tolerated, and no clinically meaningful differences in the incidence of serious adverse events, treatment-related adverse events, or discontinuations due to adverse events were observed between groups. There were no significant between-group differences in glycemic control (HbA1c) or concentrations of fasting insulin, creatine phosphokinase, or hepatic transaminases. CONCLUSION: Simvastatin was an effective and generally well tolerated treatment for hyperlipidemia when used in combination with TZD therapy in this population of patients with type 2 DM. PMID- 15110131 TI - Pharmacokinetic and pharmacodynamic properties of eptifibatide in subjects with normal or impaired renal function. AB - BACKGROUND: Previous studies of the glycoprotein IIb-IIIa inhibitor eptifibatide have included patients with moderate renal impairment (serum creatinine concentrations, 2.0-4.0 mg/dL). In these patients, adjustment of the standard infusion dose (2.0 microg/kg.min) to 1.0 microg/kg.min was recommended on empiric grounds because approximately 50% of eptifibatide is cleared renally. OBJECTIVE: The present study was designed to assess teh pharmacokinetic and pharmacodynamic properties of eptifibatide in subjects with various levels of creatinine clearance (CrCl), a parameter that is a more accurate indicator of renal function than serum creatinine concentration to determine the appropriate dose adjustment in patients with reduced renal function. A secondary objective was to determine the tolerability of eptifibatide when administered to patients with decreased renal function. METHODS: This open-label study was concluded at 3 US sites experienced in conducting similar studies. Subjects with differing renal function (normal [group 1, CrCl > 80 mL/min], mild renal impairment [group 2, CrCl 51-80 mL/min], moderate renal impairment [group 3, CrCl 30-50 mL/min], or severe renal impairment [group 4, CrCl <30 mL/min]) received a 24-hour eptifibatide infusion of 2.0 microg/kg.min (groups 1, 2, and 3) or 1.0 microg/kg.min (group 4). The primary end point was the eptifibatide steady-state plasma concentration; the secondary end points included inhibition of platelet aggregation and eptifibatide clearance, terminal plasma half-life, and area under the plasma concentration time curve. In addition to analyses performed for each group, pharmacokinetic models were estimated and eptifibatide clearance was related to CrCl as a continuous variable by linear regression. All adverse events were recorded, with particular attention to bleeding. RESULTS: Thirty-one subjects (21 men, 10 women; mean age, 58.5 years [range, 44-73 years]; mean body weight, 81.6 kg [range, 51.5 105.1 kg]; mean height, 162.9 cm [range, 150-183 cm]) were included in the study. A strong correlation was found between eptifibatide clearance and CrCl. A tree based analytic model indicated that the optimal discrete break point for the purpose of dose adjustment was a CrCl of 55.85 mL/min, which was rounded to 50 mL/min for practical clinical reasons. In patients with moderate or severe renal impairment (CrCl < or = 50 mL/min), clearance rates and steady-state concentrations of eptifibatide were approximately 50% lower and almost 2-fold higher, respectively, than in patients with normal renal function or mild renal impairment (CrCl > 50 mL/min). Inhibition of platelet aggregation exceeded the clinically significant threshold of 80% in all groups. Five subjects had a mild bleeding event and 4 subjects experienced mild to moderate nonbleeding events, 2 of which were considered drug-related by the investigator. None of the events required intervention. CONCLUSIONS: In this study, moderate to severe renal impairment was associated with an approximately 50% reduction in total eptifibatide clearance and a corresponding doubling of plasma eptifibatide concentration. Based on these findings, in patients with moderately or severely impaired renal function (calculated CrCl < or =50 mL/min) who are scheduled for percutaneous coronary intervention or who have non-ST-segment elevation acute coronary syndromes, it is appropriate to reduce the infusion dose of eptifibatide by 50% (from 2 to 1 microg/kg.min). Because the bolus dose(s) is (are) used to establish the initial targeted plasma eptifibatide concentrations, which are independent of clearance, and given that the volume of distribution did not correlate with renal function, no adjustment of the bolus dose(s) is required. PMID- 15110132 TI - A single-blind, randomized, controlled trial to assess the efficacy and tolerability of rofecoxib, diclofenac sodium, and meloxicam in patients with acute gouty arthritis. AB - BACKGROUND: Acute attacks of gouty arthritis are characterized by the rapid onset of severe pain, swelling, and erythema of the affected joint. Nonsteroidal anti inflammatory drugs are considered the drugs of first choice for treating acute gout. Rofecoxib is a specific cyclooxygenase-2 inhibitor, which has demonstrated analgesic efficacy in the setting of acute pain. Whether it is effective in the treatment of acute gouty arthritis remains to be evaluated. OBJECTIVE: The aim of this study was to assess the efficacy and tolerability of rofecoxib compared with diclofenac sodium sustained release (SR) and meloxicam in the treatment of acute gouty arthritis. METHODS: In this single-blind, randomized, controlled, parallel group study, patients aged > or =18 years with acute gout within 48 hours of onset were randomized to receive oral treatment with 2 tablets of rofecoxib (25 mg), diclofenac (75 mg), or meloxicam (7.5 mg) once daily for 7 days. The primary outcome measures were patients global assessment of response to therapy and investigator assessment of response to therapy on days 3 and 8. Other efficacy measurements included investigator assessment of total inflammatory scores on days 3 and 8 and patient assessment of pain intensity during the first 12 hours of treatment. RESULTS: Sixty-two patients (53 men, 9 women; mean [SD] age, 51.1 [12.1] years) were assigned to receive rofexocib (n = 20), diclofenac (n = 21), or meloxicam (n = 21). For patient global response to therapy on days 3 and 8, rofecoxib was associated with analgesic efficacy in significantly more patients compared with meloxicam (84.2% vs 40.0% of patients [ P=0.005] and 94.7% vs 60.0% of patients [ P=0.02], respectively); no significant differences versus diclofenac were found. Similarly, for investigator global assessment of response to therapy, a greater percentage of responders was found in the rofecoxib group compared with the meloxicam group on day 3 (88.9% vs 40.0% of patients [ P=0.02 ]), but the difference was not significant on day 8. A greater percentage of responders was found in the rofecoxib group compared with the diclofenac group on day 3 (88.9% vs 47.3% [ P=0.007 ]), but the difference was not significant on day 8. Compared with baseline, all regimens showed significant improvement in total inflammatory scores on days 3 and 8 (all P<0.01 ). During the first 12 hours after dosing, pain intensity score was significantly reduced with rofecoxib starting at 0.5 hours ( P<0.05 ), but not with diclofenac or meloxicam. Clinical adverse events (AEs) were reported in 4 (20.0%), 7 (33.3%), and 6 (28.6%) patients in the rofecoxib, diclofenac, and meloxicam groups, respectively; the most common AEs reported were edema in 1 patient each in the rofecoxib (5.0%) and meloxicam (4.8%) groups and 2 patients (9.5%) in the diclofenac group and abdominal (1 [5.0%], 1 [4.8%], and 2 [9.5%], respectively). No significant differences in tolerability were found among the 3 treatment groups. CONCLUSIONS: In this study of patients with acute gouty arthritis, rofecoxib 50 mg once daily provided more effective treatment than diclofenac sodium SR 150 mg and meloxicam 15 mg administered orally once daily for 7 days in > or = 1 efficacy assessment of overall analgesic effect on day 3 or day 8. Rofecoxib achieved a rapid onset of pain relief, demonstrating significant improvement 30 minutes after dosing. All of the regimens appeared well tolerated in the population studied. PMID- 15110133 TI - Intrahepatic arterial administration of low-dose methotrexate in patients with severe hepatic graft-versus-host disease: an open-label, uncontrolled trial. AB - BACKGROUND: Hepatic grafr-versus-host disease (GVHD) is associated with significant morbidity and mortality. Standard therapy includes systemically administered immunosuppressive drugs. More recent reports have described catheter directed intrahepatic arterial (IHA) delivery of low-dose methotrexate (MTX) and methylprednisolone in the treatment of corticosteroid-resistant severe hepatic GVHD. OBJECTIVE: This article reports on MTX toxicity and the variability in plasma drug concentrations after IHA administration of low-dose MTX in patients with severe hepatic GVHD. METHODS: In this open-label, uncontrolled pilot study, MTX and methylprednisolone were administered via the hepatic artery in patients with corticosteroid-resistant grade III or IV GVHD of the liver. Patients also received standard therapy. MTX concentrations were measured in the hepatic artery 5 and 10 minutes after injection and in peripheral venous blood at 1, 2, and 24 hours. RESULTS: Six patients (5 males [83.3%], I female [16.7%]; median age, 32 years; range, 8-42 years) were enrolled in the study. No hepatotoxicity was observed after IHA administration of MTX. In 5 patients with normal renal function, plasma drug concentrations 24 hours after administration of MTX ranged from 0.01 to 0.12 micromol/L (mean [SD], 0.043 [0.042] micromol/L). In 1 patient with renal failure, plasma MTX concentrations were 1.0 micromol/L 24 hours after administration and 0.07 micromol/L 5 days after administration. The severe hematologic and renal toxicity observed in this patient may have contributed to his death. Adverse events in patients with GVHD and normal renal function, who had normal plasma MTX concentrations, were comparable to those that have been reported after administration of an intravenous infusion. CONCLUSIONS: In patients with GVHD and normal renal function, IHA administration of low-dose MTX was not associated with liver or bone marrow toxicity. Further study is needed to determine the optimal protocols for treating corticosteroid-resistant hepatic GVHD. PMID- 15110134 TI - Public policy, private insurance benefits, and weight control. PMID- 15110135 TI - Retrospective database analysis of the prevention of venous thromboembolism with low-molecular-weight heparin in acutely III medical inpatients in community practice. AB - BACKGROUND: Clinical trials have demonstrated that prophylaxis with low-molecular weight heparin reduces the occurrence of venous thromboembolism (VTE) among acutely ill medical inpatients in the experimental setting. OBJECTIVE: The goal of this retrospective database analysis was to examine the outcomes of low molecular-weight heparin thromboprophylaxis among acutely ill medical inpatients in community practice. METHODS: Using a large, geographically diverse, multihospital US database, we identified persons aged > or =40 years who had a hospital stay > or =6 days for an acute medical condition (including selected circulatory disorders, respiratory disorders, infectious diseases, or neoplasms) during calendar-year 2000. From these patients, those who received either enoxaparin thromboprophylaxis or no thromboprophylaxis were identified. Surgical patients, patients with nonthrombotic conditions requiring anticoagulant therapy, those transferred from or discharged to another acute care facility, and those medically ineligible for anticoagulation therapy were excluded. We compared the incidence of deep-vein thrombosis (DVT), pulmonary embolism (PE), all VTE (ie, DVT and/or PE), and death during the hospital stay in the 2 cohorts. RESULTS: A total of 162 patients receiving enoxaparin thromboprophylaxis and 3557 receiving no thromboprophylaxis were identified. The risk of VTE over the course of hospitalization was 1.9% with enoxaparin thromboprophylaxis versus 6.2% with no thromboprophylaxis (relative risk = 0.30; P=0.023 ); mortality was similar in the 2 groups (8.0% vs 7.3; P=NS ). CONCLUSIONS: Using hospital administrative data, we observed a 70% lower risk of VTE for hospitalized acutely ill medical patients receiving low-molecular-weight heparin thromboprophylaxis verus those receiving no thromboprophylaxis; these results are consistent with findings from clinical trials of low-molecular-weight heparin versus placebo. We conclude that the low molecular-weight heparin enoxaparin is effective in reducing the risk of VTE in acutely ill medical inpatients in community practice. PMID- 15110136 TI - Canadian economic comparison of extended-release oxybutynin and immediate-release tolterodine in the treatment of overactive bladder. AB - BACKGROUND: Overactive bladder (OAB) is a condition characterized by urgency, increased frequency of micturition, or urge incontinence. It affects a considerable segment of the population, particularly with increasing age. Pharmacotherapy is one of the most common approaches to the treatment of OAB. OBJECTIVE: This article describes the development and results of a model comparing health-economic outcomes for the new extended-release (XL) formulation of oxybutynin and immediate-release (IR) tolterodine in a population of community dwelling Canadian adults with OAB. METHODS: A Markov model was developed to compare health-economic outcomes over the course of 1 year. Effectiveness and treatment-persistence data were derived from the OBJECT (Overactive Bladder: Judging Effective Control and Treatment) trial, a 3-month comparison of oxybutynin XL 10 mg and tolterodine IR 4 mg, and were used, together with data from the literature (identified through a MEDLINE search of articles published between 1990 and 2003), to project outcomes beyond the trial period. Severity specific cost profiles for incontinence were developed. In the principal analyses, cost items were limited to drug therapy, physician visits, use of pads or other protection, and laundry costs. Costs are reported in 2002 Canadian dollars. RESULTS: Costs after 1 year were estimated to be an average of $32 less per patient for oxybutynin XL compared with tolterodine IR, and 3.1 additional patients in every 100 who received oxybutynin XL were expected to attain complete continence compared with those who received tolterodine. During the course of 1 year, patients receiving oxybutynin XL were expected to have a mean 16.5 additional incontinence-free days compared with those receiving tolterodine IR. The results were sensitive to relative drug prices. In the other sensitivity analyses, however, oxybutyrin XL maintained its advantage over a wide range of inputs. CONCLUSION: The results of these analyses suggest that when priced equivalently, oxybutynin XL would reduce costs and provide better results than tolterodine IR over 1 year of treatment. PMID- 15110137 TI - Underuse of aspirin in type 2 diabetes mellitus: prevalence and correlates of therapy in rural Canada. AB - BACKGROUND: Patients with type 2 diabetes mellitus (DM) have a markedly increased risk of cardiovascular morbidity and mortality. Guidelines of both the American and Canadian Diabetes Associations recommend the use of aspirin as antiplatelet therapy for all adults with type 2 DM. OBJECTIVES: The aims of this study were to assess the rate of adherence to guidelines for aspirin use in DM patients in rural Canadian communities and to describe the independent correlates of aspirin use in this population. METHODS: We collected information from a cohort of patients with type 2 DM living in 2 rural regions of northern Alberta, Canada, at the time of their enrollment in a multidisciplinary outreach program designed to improve their quality of care. Our primary outcome was self-reported use of antiplatelet therapy (aspirin or others). We use multivariate logistic regression analyses to examine the independent association between sociodemographic and clinical characteristics and self-reported use of antiplatelet agents. RESULTS: Among 342 patients included in the study (who were typical of rural Canadian patients with type 2 DM), the mean age was 62.9 years; 149 (44%) were men, 84 (25%) were of indigenous origin, and the median time since diagnosis of DM was 8 years. Despite guideline recommendations, only 23% of the cohort (78 patients) were regularly taking aspirin alone or in combination with a thienopyridine (n = 74 and n = 2, respectively) or a thienopyridine alone (n = 2). The results of them ultivariate analyses showed that the only factors independently associated with the use of antiplatelet therapy were symptomatic coronary artery disease (adjusted odds ratio [AOR], 3.1; 95% CI, 1.1-8.7; P=0.033 ), older age (AOR, 2.0 per 10-year interval; 95% CI, 1.7-2.2; P<0.001 ); and male sex (AOR, 1.9; 95% CI, 1.1-3.5; P=0.026 ). CONCLUSIONS: Aspirin is a safe, inexpensive, and readily available therapy that is effective for preventing cardiovascular disease, and patients with type 2 DM are particularly likely to benefit from such preventive therapy. However, we found significant underuse of aspirin therapy among our study population. Aspirin should be included and better promoted as a factor in high-quality, evidence-based DM management. PMID- 15110138 TI - Two centuries of excitation-contraction coupling. PMID- 15110139 TI - A cell physiologist between East and West Germany. PMID- 15110140 TI - Functional architecture of the SR calcium store in uterine smooth muscle. AB - Sarcoplasmic reticulum (SR) is abundant in uterine smooth muscle cells. The functional role of this organelle in the regulation of uterine myocytes is not fully understood. The data available in the literature suggest that SR plays a dual role: as a source of calcium and as a calcium sink shaping calcium transients produced by membrane depolarisation and uterotonic agonists. Advances in digital imaging techniques including confocal microscopy of isolated living cells, and the development of methods for direct measurement of intraluminal calcium, has triggered a substantial increase in the number of publications elucidating the role of intracellular stores in calcium signalling. In this paper we review the literature and our own work on the SR calcium store in uterine smooth muscle cells. PMID- 15110141 TI - Vascular smooth muscle mitochondria at the cross roads of Ca(2+) regulation. AB - Mitochondria play an essential role in the regulation of vascular smooth muscle Ca(2+) signaling being simultaneously integrated in the regulation of ion channels and Ca(2+) transporters, oxygen radical production, metabolite recycling and intracellular redox potential. Mitochondria buffer Ca(2+) from cytoplasmic microdomains to alter the spatio-temporal pattern of Ca(2+) gradients following Ca(2+)-influx and Ca(2+)-release, and thus control site-specific, Ca(2+) dependent ion channel activation and inactivation. The sub-cellular localization of mitochondria in conjunction with tissue-specific channel expression is fundamental to vascular heterogeneity. The mitochondrial electron transport chain recycles metabolic intermediates that modulate cellular redox potential and produces oxygen radicals in proportion to oxygen tension. Perturbation of specific complexes within the transport chain can affects NADH:NAD and ATP:ADP ratios and radical production, which can in turn influence second messenger metabolism, ion channel gating and Ca(2+)-transporter activity. Mitochondria thus provide the common ground for cross-talk between these regulatory systems that are mutually sensitive to one another. This cross-talk between signaling systems provides a means to render the physiological regulation of vascular tone responsive to complex stimulation by paracrine and endocrine factors, blood pressure and flow, tissue oxygenation and metabolic state. PMID- 15110142 TI - Imaging calcium entering the cytosol through a single opening of plasma membrane ion channels: SCCaFTs--fundamental calcium events. AB - Recently, it has become possible to record the localized fluorescence transient associated with the opening of a single plasma membrane Ca(2+) permeable ion channel using Ca(2+) indicators like fluo-3. These Single Channel Ca(2+) Fluorescence Transients (SCCaFTs) share some of the characteristics of such elementary events as Ca(2+) sparks and Ca(2+) puffs caused by Ca(2+) release from intracellular stores (due to the opening of ryanodine receptors and IP(3) receptors, respectively). In contrast to intracellular Ca(2+) release events, SCCaFTs can be observed while simultaneously recording the unitary channel currents using patch-clamp techniques to verify the channel openings. Imaging SCCaFTs provides a way to examine localized Ca(2+) handling in the vicinity of a channel with a known Ca(2+) influx, to obtain the Ca(2+) current passing through plasma membrane cation channels in near physiological solutions, to localize Ca(2+) permeable ion channels on the plasma membrane, and to estimate the Ca(2+) currents underlying those elementary events where the Ca(2+) currents cannot be recorded. Here we review studies of these fluorescence transients associated with caffeine-activated channels, L-type Ca(2+) channels, and stretch-activated channels. For the L-type Ca(2+) channel, SCCaFTs have been termed sparklets. In addition, we discuss how SCCaFTs have been used to estimate Ca(2+) currents using the rate of rise of the fluorescence transient as well as the signal mass associated with the total fluorescence increase. PMID- 15110143 TI - Intracellular Ca(2+) dynamics and sarcomere length in single ventricular myocytes. AB - The measurements of the sarcomere length in dissociated cardiac ventricular myocytes are discussed using mainly our own experimental data. The striation periodicity of these unloaded cells was found to be that which is to be expected of a myocyte free of the ultrastructural constraints imposed upon it by the normal syncytial matrix of the ventricular wall. The sarcomere length and [Ca(2+)] relationship was consistent as expected from the intact tissue, when it was measured soon after partial rupturing the cell membrane. Miniature fluctuations of individual sarcomere length were demonstrated during rest, which was augmented by the Ca(2+) overload. The [Ca(2+)] could be estimated from the measurements of sarcomere length during the positive staircase of contraction. The usefulness of the optical measurement of sarcomere pattern was indicated. PMID- 15110144 TI - Drugs, hERG and sudden death. AB - Early recognition of potential QT/TdP liability is now an essential component of the drug discovery/drug development program. The hERG assay is an indispensable step and a high-quality assay must accompany any investigational new drug (IND) application. While it is the gold standard at present, the hERG assay is too labor-intensive and too low throughput to be used as a screen early in the discovery/development process. A variety of indirect high throughput screens have been used. PMID- 15110145 TI - Tonic component of myocardial contraction. AB - Calcium transients and contractions of cardiac myocytes consist of phasic component, relaxing spontaneously independently of membrane voltage and of the tonic component (TC) relaxing only upon repolarization. Experimental data reviewed in this article suggest that most Ca(2+) activating TC is released from sarcoplasmic reticulum (SR) via the ryanodine receptors (RyRs). Most likely these RyRs are activated by sustained Ca(2+) influx. However, its route may differ depending on species and state of the cells. It seems that in rat RyRs responsible for TC are activated by the sustained Ca(2+) current. In guinea-pig the blockers of Ca(2+) current or reverse mode Na(+)/Ca(2+) exchange do not inhibit TC, so these routes seem unlikely. In myocytes of the failing human hearts TC is activated mostly via the reverse mode Na(+)/Ca(2+) exchange and contribution of SR is negligible. The mechanism of TC in the normal human cardiomyocytes has not been investigated. Thus, despite investigation of TC for half a century many problems concerning the mechanism of its activation and maintenance as well as its physiological meaning remain unsolved. PMID- 15110146 TI - Intracellular Ca(2+) concentration and rate adaptation of the cardiac action potential. AB - Influx of Ca(2+) ions through the cardiac plasma membrane contributes to the shaping of the action potential plateau and acts as trigger for the release of Ca(2+) ions from the sarcoplasmic reticulum and the initiation of the contractile process. The increased intracellular Ca(2+) concentration feeds back on the channels and transporters in the plasma membrane and modulates the electrical activity. This interaction and its change with rate of pacing is the topic of this review, which is subdivided in three parts. In part I a description is given of different channels and transporters that carry Ca(2+) ions, or are activated modulated by intracellular Ca(2+) ions. In part II an analysis is given of the changes in action potential duration and shape when stimuli are applied in the relative refractory period (electrical restitution) and when rate is suddenly increased and kept at the higher level until steady-state is obtained. A description of experimental findings in each case is followed by a discussion of possible mechanisms. Part III deals with physiopathological aspects of Ca(2+) handling and discusses recent information on hypertrophy, heart failure and atrial fibrillation. PMID- 15110147 TI - Pharmacological modulation of ion channels and transporters. AB - Ion channels and transporter proteins are prerequisites for formation and conduction of cardiac electrical impulses. Acting in concert, these proteins maintain cellular Na(+) and Ca(2+) homeostasis. Since intracellular Ca(2+) concentration determines contractile activation, we expect the majority of agents that modulate activity of ion channels and transporters not only to influence cellular action potentials but also contractile force. Drugs which block ion channels usually possess antiarrhythmic properties, those inhibiting the Na(+) pump have predominantly inotropic effects and those affecting Na(+),Ca(2+)- or Na(+),H(+)-exchanger protect against ischaemic cell damage. However, irrespective of their primary indication, all compounds targeted against ion channels and transporter proteins possess potential proarrhythmic activity. PMID- 15110148 TI - Physiological and pathological modulation of ryanodine receptor function in cardiac muscle. AB - Calcium release from the sarcoplasmic reticulum (SR) in cardiac muscle occurs through a specialised release channel, the ryanodine receptor, RyR, via the process of Ca-induced Ca release (CICR). The open probability of the RyR is increased by elevation of cytoplasmic Ca concentration ([Ca(2+)](i)). However, in addition to Ca, other modulators affect the RyR open probability. Agents which increase the RyR opening during systole produce a transient increase of systolic [Ca(2+)](i) followed by a return to the initial level due to a compensating decrease of SR Ca content. Increasing RyR opening during diastole decreases SR Ca content and thereby decreases systolic [Ca(2+)](i). We therefore conclude that potentiation of RyR opening will, if anything, decrease systolic [Ca(2+)](i). The effects of specific examples of modulators of the RyR, such as phosphorylation, metabolic changes, heart failure and polyunsaturated fatty acids, are discussed. PMID- 15110149 TI - Putting out the fire: what terminates calcium-induced calcium release in cardiac muscle? AB - The majority of contractile calcium in cardiac muscle is released from stores in the sarcoplasmic reticulum (SR), by a process of calcium-induced calcium release (CICR) through ryanodine receptors. Because CICR is intrinsically self reinforcing, the stability of and graded regulation of cardiac EC coupling appear paradoxical. It is now well established that this gradation results from the stochastic recruitment of varying numbers of elementary local release events, which may themselves be regenerative, and which can be directly observed as calcium sparks. Ryanodine receptors (RyRs) are clustered in dense lattices, and most calcium sparks are now believed to involve activation of multiple RyRs. This implies that local CICR is regenerative, requiring a mechanism to terminate it. It was initially assumed that this mechanism was inactivation of the RyR, but during the decade since the discovery of sparks, no sufficiently strong inactivation mechanism has been demonstrated in vitro and all empirically determined gating schemes for the RyR give unstable EC coupling in Monte Carlo simulations. We consider here possible release termination mechanisms. Stochastic attrition is the spontaneous decay of active clusters due to random channel closure; calculations show that it is much too slow unless assisted by another process. Calcium-dependent RyR inactivation involving third-party proteins remains a viable but speculative mechanism; current candidates include calmodulin and sorcin. Local depletion of SR release terminal calcium could terminate release, however calculations and measurements leave it uncertain whether a sufficient diffusion resistance exists within the SR to sustain such depletion. Depletion could be assisted by dependence of RyR activity on SR lumenal [Ca(2+)]. There is substantial evidence for such lumenal activation, but it is not clear if it is a strong enough effect to account for the robust termination of sparks. The existence of direct interactions among clustered RyRs might account for the discrepancy between the inactivation properties of isolated RyRs and intact clusters. Such coupled gating remains controversial. Determining the mechanism of release termination is the outstanding unsolved problem of cardiac EC coupling, and will probably require extensive genetic manipulation of the EC coupling apparatus in its native environment to unravel the solution. PMID- 15110150 TI - [Na(+)] in the subsarcolemmal 'fuzzy' space and modulation of [Ca(2+)](i) and contraction in cardiac myocytes. AB - The strength of the heart beat depends on the amplitude and time course of the transient increase in [Ca(2+)] in the myocytes with each cycle. [Na(+)](i) modulates cardiac contraction through its effect on the Ca(2+) flux through the Na/Ca exchanger. Cardiac excitation-contraction coupling has been postulated to occur in a microdomain or 'fuzzy' space at the junction of the T-tubules and the sarcoplasmic reticulum. This 'fuzzy' space is well described for the Ca(2+) fluxes and the interaction between the L-type Ca(2+) channel, the Ca(2+) release channel of the sarcoplasmic reticulum and the Na/Ca exchanger. Co-localization of the Na(+) transporters, in particular the Na/K pump and the Na(+) channel, within this 'fuzzy' space is not as well established. The functional and morphological characteristics of the 'fuzzy' space for Na(+) and its interaction with the Ca(2+) handling suggest that this space is not strictly co-inciding with the Ca(2+) microdomain. In this space [Na(+)] can be several-fold higher or lower than [Na(+)] in the bulk cytosol. This has implications for modulation of [Ca(2+)](i) during a single beat as well as during alterations in Na(+) fluxes seen in pathological conditions. PMID- 15110151 TI - Interactions between inositol 1,4,5-trisphosphate receptors and ryanodine receptors in smooth muscle: one store or two? AB - This short review proposes a system of simplified functional models describing possible interactions between Ca(2+)-release channels associated with IP(3)Rs and RyRs in smooth muscle, and considers each of these models in the light of the available experimental evidence. Complete separation of IP(3)R- and RyR-gated stores seems to be unusual. Where both receptors release Ca(2+) from a common pool, simple interactions can occur since changes in the activation of one receptor type affects the availability of Ca(2+) for release through the other. Alterations in [Ca(2+)] within the sarcoplasmic reticulum can also affect the open probability of the release channels, and not just the Ca(2+)-flux through the channels when open, e.g., Ca(2+)-release through tonically active IP(3)Rs appears to limit SR Ca(2+)-content in some myocytes, and this modulates RyR activity, as indicated by changes in Ca(2+)-spark frequency. There is also evidence that intracellular release channels may co-operate, leading to positive feedback during activation. In particular, agonist-dependent activation of IP(3)Rs can promote activation of RyRs, amplifying and shaping the resulting Ca(2+)-signal. While there is little direct evidence as to the mechanism responsible for this interaction, some form of Ca(2+)-induced Ca(2+)-release in response to local increases in [Ca(2+)](c) seems likely. PMID- 15110152 TI - Molecular regulation of cardiac ryanodine receptor ion channel. AB - The release of Ca(2+) ions from intracellular stores is a key step in a wide variety of cellular functions. In striated muscle, the release of Ca(2+) from the sarcoplasmic reticulum (SR) leads to muscle contraction. Ca(2+) release occurs through large, high-conductance Ca(2+) release channels, also known as ryanodine receptors (RyRs) because they bind the plant alkaloid ryanodine with high affinity and specificity. The RyRs are isolated as 30S protein complexes comprised of four 560 kDa RyR2 subunits and four 12 kDa FK506 binding protein (FKBP12) subunits. Multiple endogenous effector molecules and posttranslational modifications regulate the RyRs. This review focuses on current research toward understanding the control of the isolated cardiac Ca(2+) release channel/ryanodine receptor (RyR2) by Ca(2+), calmodulin, thiol oxidation/reduction and nitrosylation, and protein phosphorylation. PMID- 15110153 TI - Beyond Bowditch: the convergence of cardiac chronotropy and inotropy. AB - The ability of the heart to acutely beat faster and stronger is central to the vertebrate survival instinct. Released neurotransmitters, norepinephrine and epinephrine, bind to beta-adrenergic receptors (beta-AR) on pacemaker cells comprising the sinoatrial node, and to beta-AR on ventricular myocytes to modulate cellular mechanisms that govern the frequency and amplitude, respectively, of the duty cycles of these cells. While a role for sarcoplasmic reticulum Ca(2+) cycling via SERCA2 and ryanodine receptors (RyR) has long been appreciated with respect to cardiac inotropy, recent evidence also implicates Ca(2+) cycling with respect to chronotropy. In spontaneously beating primary sinoatrial nodal pacemaker cells, RyR Ca(2+) releases occurring during diastolic depolarization activate the Na(+)-Ca(2+) exchanger (NCX) to produce an inward current that enhances their diastolic depolarization rate, and thus increases their beating rate. beta-AR stimulation synchronizes RyR activation and Ca(2+) release to effect an increased beating rate in pacemaker cells and contraction amplitude in myocytes: in pacemaker cells, the beta-AR stimulation synchronization of RyR activation occurs during the diastolic depolarization, and augments the NCX inward current; in ventricular myocytes, beta-AR stimulation synchronizes the openings of unitary L-type Ca(2+) channel activation following the action potential, and also synchronizes RyR Ca(2+) releases following depolarization, and in the absence of depolarization, both leading to the generation of a global cytosolic Ca(i) transient of increased amplitude and accelerated kinetics. Thus, beta-AR stimulation induced synchronization of RyR activation (recruitment of additional RyRs to fire) and of the ensuing Ca(2+) release cause the heart to beat both stronger and faster, and is thus, a common mechanism that links both the maximum achievable cardiac inotropy and chronotropy. PMID- 15110154 TI - Smooth muscle cells and interstitial cells of blood vessels. AB - A rise in intracellular ionised calcium concentration ([Ca(2+)](i)) at sites adjacent to the contractile proteins is a primary signal for contraction in all types of muscles. Recent progress in the development of imaging techniques with special accent on the fluorescence confocal microscopy and new achievements in the synthesis of organelle- and ion-specific fluorochromes provide an experimental basis for study of the relationship between the structural organisation of the living smooth muscle myocyte and the features of calcium signalling at subcellular level. Applying fluorescent confocal microscopy and tight-seal recording of transmembrane ion currents to freshly isolated vascular myocytes we have demonstrated that: (1) Ca(2+) sparks originate from clustered opening of ryanodine receptors (RyRs) and build up a cell-wide increase in [Ca(2+)](i) upon myocyte excitation; (2) spontaneous Ca(2+) sparks occurred at the highest rate at certain preferred locations, frequent discharge sites (FDS), which are associated with a prominent portion of the sarcoplasmic reticulum (SR) located close to the cell membrane; (3) Ca(2+)-dependent K(+) and Cl(-) channels sense the local changes in [Ca(2+)](i) during a calcium spark and thereby couple changes in [Ca(2+)](i) within a microdomain to changes in the membrane potential, thus affecting excitability of the cell; (4) an intercommunication between RyRs and inositol trisphosphate receptors (IP(3)Rs) is one of the important determinants of intracellular calcium dynamics that, in turn, can modulate the cell membrane potential through differential targeting of calcium dependent membrane ion channels. Furthermore, using immunohystochemical approaches in combination with confocal imaging we identified non-contractile cells closely resembling interstitial cells (ICs) of Cajal (which are considered to be pacemaker cells in the gut) in the wall of portal vein and mesenteric artery. Using electron microscopy, tight-seal recording and fluorescence confocal imaging we obtained information on the morphology of ICs and their possible coupling to smooth muscle cells (SMCs), calcium signalling in ICs and their electrophysiological properties. The functions of these cells are not yet fully understood; in portal vein they may act as pacemakers driving the spontaneous activity of the muscle; in artery they may have other a yet unsuspected functions. PMID- 15110155 TI - The postconcussion syndrome and whiplash injuries: a question-and-answer review for primary care physicians. PMID- 15110156 TI - Neck pain. PMID- 15110157 TI - Low back pain. PMID- 15110158 TI - Entrapment neuropathies. PMID- 15110159 TI - Peripheral nerve disorders. PMID- 15110160 TI - Seizure disorders. PMID- 15110161 TI - Clinical diagnosis of patients with cerebrovascular disease. PMID- 15110162 TI - Movement disorders. PMID- 15110163 TI - Memory complaints and dementia. PMID- 15110164 TI - A discussion of sleep. PMID- 15110165 TI - Syncope and orthostatic intolerance for the primary care physician. PMID- 15110166 TI - Multiple sclerosis. PMID- 15110168 TI - Specificity of molecular recognition learned from the crystal structures of TRAIL and the TRAIL:sDR5 complex. AB - TRAIL is a member of the tumor necrosis factor (TNF) superfamily. TRAIL has drawn a lasting attention because of its selectivity and efficacy in inducing apoptosis in a variety of cancer cells but not in normal cells. The structures of both TRAIL and the protein in complex with the extracellular domain of death receptor 5 (sDR5) were elucidated. Because each factor of the ligand family and the receptor family is large, it poses an intriguing question of how recognition between cognate ligands and receptors is achieved in a highly specific manner without cross interactions. This review focuses on the unique properties of TRAIL and molecular strategies for the specific recognition between the two family members primarily based on the crystal structures of TRAIL and the TRAIL:sDR5 complex. PMID- 15110169 TI - Crystal structure of RANK ligand involved in bone metabolism. AB - Bone remodeling involves the resorption of bone by osteoclasts and the synthesis of bone matrix by osteoblasts. Recently, an essential cytokine system for osteoclast biology has been identified and extensively characterized. This system consists of a ligand, receptor activator of NF-kappaB ligand (RANKL), a receptor, RANK, and its soluble decoy receptor, osteoprotegerin (OPG). RANKL, a member of the tumor necrosis factor (TNF) family, triggers osteoclastogenesis by forming a complex with RANK, a member of the TNF receptor family. Because members of the TNF family have the same topology and the extracellular domains of the TNF receptor family members also adopt the same structural scaffold, in addition to their rapid increase in the number, this poses an intriguing question of how recognition between cognate ligands and receptors is achieved in a highly specific manner. Structural studies on the mouse RANKL extracellular domain showed that the RANKL is trimeric, and each subunit has a beta-strand jellyroll topology like the other members of the TNF family. A comparison of RANKL with TNF beta and TNF-related apoptosis-inducing ligand (TRAIL), whose structures were determined to be in the complex form with their respective receptor, revealed conserved and specific features of RANKL in the TNF superfamily. Residues important for receptor binding and activation have also been confirmed by mutagenesis experiments. Further structural and mutational studies on the RANKL/RANK/OPG system will provide useful information for developing drug candidates that inhibit osteoclastogenesis and mediate problems of bone metabolism. PMID- 15110170 TI - Promoter of TRAIL-R2 gene. AB - TRAIL-R2 promoter does not have a typical TATA-box but two functional Sp1-binding sites. TRAIL-R2 promoter belongs to the class of TATA-less and GC-box-containing promoters. The minimal promoter element is contained in the region spanning -198 to -116 upstream of translational initiation codon ATG. Computer analysis shows putative transcription factor binding sites such as c-Ets, AML-1a, c-Myb, Sp1, and GATA-1 in TRAIL-R2 promoter. Hypermethylation of TRAIL-R2 is not frequent compared with that of TRAIL-R3 and TRIAL-R4. There are no potential transcription factor binding sites in highly homologous regions between TRAIL-R2 promoter and TRAIL-R1 promoter, or between TRAIL-R2 promoter and mouse homologue mouse killer (MK) promoter. TRAIL-R2 is known to be a downstream gene of p53, a tumor suppressor gene, and a p53-binding site in TRAIL-R2 intron 1 is responsible for p53-dependent transcription. Thapsigargin, endoplasmic reticulum Ca(2+)-ATPase inhibitor calcium releaser, upregulates TRAIL-R2 expression via the promoter region. Many regulators of TRAIL-R2 have been reported. However, it has not been demonstrated whether they regulate TRAIL-R2 via the promoter region. Here, we show a list of these regulators. Finally, we demonstrate the possibility of cancer therapy using regulation of TRAIL-R2 promoter. PMID- 15110171 TI - Transcriptional regulation of the TRAIL-R3 gene. AB - TRAIL-R3 is a decoy receptor for TRAIL (tumor necrosis factor-related apoptosis inducing ligand), a member of the tumor necrosis factor (TNF) ligand family. TRAIL induces apoptosis in a broad range of cancer cell lines, but not in many normal cells-a finding that generated extraordinary excitement about its potential as a specific antitumor agent. In several cell types, decoy receptors inhibit TRAIL-induced apoptosis by binding to it and preventing its binding to TRAIL proapoptotic or death receptors. However, recently published data regarding the role of these receptors in TRAIL-induced cellular death are contradictory. The key to resolving this controversy may lie in the regulation and cellular localization of TRAIL receptors. In this regard, cloning and analysis of the TRAIL-R3 promoter will help to identify the cellular factors that regulate its transcriptional expression. This chapter summarizes current knowledge in this field and outlines directions for future research. PMID- 15110172 TI - Monoclonal antibodies against TRAIL. AB - TRAIL (tumor necrosis factor related apoptosis inducing ligand) is a cytokine proposed to be used in cancer therapy, since it kills cancer cells but not normal cells. Also, recent studies report that TRAIL inhibits the development of arthritis. In order to investigate the role of TRAIL in health and disease, monoclonal antibodies against TRAIL have been developed. This chapter gives an overview of different monoclonal antibodies against TRAIL which are published or commercially available. Monoclonal antibodies against TRAIL are useful in different immunological techniques, and this chapter presents an overview of the applications of these antibodies with a focus on immunoassays for detection of soluble TRAIL. In addition, the physiological significance of some results obtained by using monoclonal antibodies against TRAIL are discussed. PMID- 15110173 TI - Modulation of TRAIL signaling complex. PMID- 15110174 TI - TRAIL and NFkappaB signaling--a complex relationship. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) or Apo2L is a ligand of the TNF family interacting with five different receptors of the TNF receptor superfamily, including two death receptors. It has attracted wide interest as a potential anticancer therapy because some recombinant soluble forms of TRAIL induce cell death predominantly in transformed cells. The nuclear factor kappaB (NFkappaB)?Rel family of proteins are composed of a group of dimeric transcription factors that have an outstanding role in the regulation of inflammation and immunity. Control of transcription by NFkappaB proteins can be of relevance to the function of TRAIL in three ways. First, induction of antiapoptotic NFkappaB dependent genes critically determines cellular susceptibility toward apoptosis induction by TRAIL-R1, TRAIL-R2, and other death receptors. Each of the multiple of known NFkappaB inducers therefore has the potential to interfere with TRAIL-induced cell death. Second, TRAIL and some of its receptors are inducible by NFkappaB, disclosing the possibility of autoamplifying TRAIL signaling loops. Third, the TRAIL death receptors can activate the NFkappaB pathway. This chapter summarizes basic knowledge regarding the understanding of the NFkappaB pathway and focuses on its multiple roles in TRAIL signaling. PMID- 15110175 TI - CARDINAL roles in apoptosis and NFkappaB activation. AB - Numerous proteins containing the caspase recruitment domain (CARD) have now been identified. While certain CARD-containing proteins are involved in caspase activation in the context of apoptosis, many others participate in NFkappaB signaling pathways associated with innate or adaptive immune responses. Here, we discuss the CARD-containing proteins that have been implicated as participants in immune response signaling pathways that culminate in NFkappaB activation. PMID- 15110176 TI - TRAIL in the airways. AB - Tumor necrosis factor-related apoptosis inducing ligand (TRAIL) is an important immunomodulatory factor that may play a role in the structural changes observed in the asthmatic airways. In vitro as well as in vivo studies have evidenced a dual role for TRAIL: it can either function as a pro- or anti-inflammatory cytokine on inflammatory cells, participating in the initiation and resolution of inflammatory and immune responses. TRAIL is expressed in the airways by inflammatory cells infiltrated in the bronchial mucosa, as well as by structural cells of the airway wall including fibroblasts, epithelial, endothelial, and smooth muscle cells. By releasing TRAIL, these different cell types may then participate in the increased levels of TRAIL observed in bronchoalveolar lavage fluid from asthmatic patients. Taken together, this suggests that TRAIL may play a role in inflammation in asthma. However, concerning its role is dual in the modulation of inflammation, further studies are needed to elucidate the precise role of TRAIL in the airways. PMID- 15110177 TI - TRAIL death receptors, Bcl-2 protein family, and endoplasmic reticulum calcium pool. AB - Calcium (Ca(2+)) is one of the highly versatile second messengers critical in cellular pathophysiology. Alterations in Ca(2+) homeostasis affect many cellular processes, including apoptosis. Recent studies have started to unravel the molecular mechanisms of apoptosis regulation in context to intracellular Ca(2+) pools. In this regard, Bcl-2 has been reported to mediate its anti-apoptotic effects, partly, by lowering the endoplasmic reticulum (ER) Ca(2+) load and by inhibiting the mitochondrial uptake of Ca(2+). However, the opposite is true for Bax and Bak that promote apoptosis, in part, by increasing the ER Ca(2+) load and Ca(2+) transfer from the ER to mitochondria. Massive ER Ca(2+) depletion coupled with upregulation of DR5 has also been reported to induce apoptosis. The mechanistic details of how some of these molecules affect intracellular Ca(2+) contents and sense perturbations in Ca(2+) homeostasis remain to be elucidated. The recent explosion of information in the fields of cell signaling and apoptosis is likely to facilitate the future investigations aiming to explore these issues. PMID- 15110178 TI - FLIP protein and TRAIL-induced apoptosis. AB - Death ligands (such as Fas/CD95 ligand and TRAIL?Apo2L) and death receptors (such as Fas/CD95, TRAIL-R1?DR4, and TRAIL-R2/DR5) are involved in immune-mediated neutralization of activated or autoreactive lymphocytes, virus-infected cells, and tumor cells. Consequently, dysregulation of death receptor-dependent apoptotic signaling pathways has been implicated in the development of autoimmune diseases, immunodeficiency, and cancer. Moreover, the death ligand TRAIL has gained considerable interest as a potential anticancer agent, given its ability to induce apoptosis of tumor cells without affecting most types of untransformed cells. The FLICE-inhibitory protein (FLIP) potently blocks TRAIL-mediated cell death by interfering with caspase-8 activation. Pharmacologic down-regulation of FLIP might serve as a therapeutic means to sensitize tumor cells to apoptosis induction by TRAIL. PMID- 15110179 TI - Epidermal growth factor and trail interactions in epithelial-derived cells. AB - In healthy tissues, there is a balance between cell survival and death. This balance ensures epithelial cells survive in the right milieu, but undergo programmed cell death (apoptosis) when the environment is no longer supportive. Cells sense these changes primarily through receptors on the cell surface that bind to specific ligands present in the extracellular environment. These receptors, through signal transduction pathways, lead to promotion of cell survival or induction of cell death. One of the most important types of receptors regulating cell survival is the epidermal growth factor (EGF) receptors, while one of the most important types of receptors regulating apoptosis is the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) death receptors. EGF receptors activate survival signaling pathways including PI3K/AKT, Ras/MAPK, and JAK/STAT signaling pathways leading to cell survival. TRAIL activates apoptotic signaling pathways leading to caspase activation and mitochondrial dysfunction. The balance between these two signaling pathways determine whether a cell survives or dies. In disease states, this balance is altered. For example, epithelial-derived cancer cells often have increased expression of EGF receptors and are resistant to apoptosis. Understanding the interactions between survival and apoptotic signaling pathways mediated by EGF receptors and TRAIL death receptors will be essential to explain the role these pathways play in healthy and diseased cells. PMID- 15110181 TI - TRAIL and viral infection. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF family that can induce apoptosis when binding to either of two receptors bearing an intracellular death domain. The physiologic function of the TRAIL system, which also comprises three receptors not mediating a death signal has just begun to be elucidated. Expression of TRAIL, mostly upon stimulation by interferons, in different cytotoxic immune cells suggested it has a role as an important effector molecule in immune surveillance. In addition to its ability to induce apoptosis in transformed tumor cells, TRAIL has attracted attention for its possibly critical role in the defense against viral infection. Viruses may induce TRAIL expression in host and?or immune cells and sensitize host cells toward TRAIL-mediated apoptosis. On the other hand, viruses have evolved a variety of strategies to prevent TRAIL-mediated host cell death early in infection, which may contribute to allowing their replication and the spread of viral progeny. The knowledge of the molecular mechanisms leading to modification of TRAIL sensitivity in virus-host cell interactions may also impact upon future (virus-based) strategies to increase TRAIL sensitivity of tumor cells. PMID- 15110180 TI - TRAIL and ceramide. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a clinically useful cytokine. TRAIL induces apoptosis in a wide variety of transformed cells, but does not cause toxicity to most normal cells. Recent studies show that death receptors (DR4 and DR5), decoy receptors (DcR1 and DcR2), and death inhibitors (FLIP, FAP-1, and IAP) are responsible for the differential sensitivity to TRAIL of normal and tumor cells. Several researchers have also shown that genotoxic agents, such as chemotherapeutic agents and ionizing radiation, enhance TRAIL induced cytotoxicity by increasing DR5 gene expression or decreasing the intracellular level of FLIP, an antiapoptotic protein. Previous studies have shown that ceramide helps to regulate a cell's response to various forms of stress. Stress-induced alterations in the intracellular concentration of ceramide occur through the activation of a variety of enzymes that synthesize or catabolize ceramide. Increases in intracellular ceramide levels modulate apoptosis by acting through key proteases, phosphatases, and kinases. This review discusses the interaction between TRAIL and ceramide signaling pathways in regulating apoptotic death. PMID- 15110182 TI - Modulation of TRAIL signaling for cancer therapy. AB - Apoptosis, the cell's intrinsic death program, is a key regulator of tissue homeostasis, and an imbalance between cell death and proliferation may result in tumor formation. Also, killing of cancer cells by cytotoxic therapies such as chemotherapy, gamma-irradiation or ligation of death receptors is predominantly mediated by triggering apoptosis in target cells. Tumor necrosis factor-related apoptosis-inducing ligans (TRAIL) is a member of the tumor necrosis factor (TNF) superfamily that induces apoptosis upon binding to its receptors. TRAIL is of special interest for cancer therapy, because TRAIL has been shown to predominantly kill cancer cells, while sparing normal cells. Importantly, combined treatment with TRAIL together with chemotherapy or gamma-irradiation synergized to achieve antitumor activity in tumor cell lines and also in tumor models in vivo. However, failure to undergo apoptosis in response to TRAIL treatment may result in tumor resistance. Understanding the molecular events that regulate TRAIL-induced apoptosis and their deregulation in resistant forms of cancer may provide new opportunities for cancer therapy. Thus, novel strategies targeting tumor cell resistance will be based on further insights into the molecular mechanisms of cell death, e.g., triggered by TRAIL. PMID- 15110183 TI - Interferon-gamma and TRAIL in human breast tumor cells. AB - Induction of apoptosis in tumor cells by death receptor activation is a novel therapeutic strategy. However, in systemic antitumor treatments, severe toxic effects have been observed with tumor necrosis factor-alpha (TNF-alpha) and CD95 ligand. TNF-alpha causes a lethal inflammatory response and CD95L produces lethal liver damage. Preclinical studies in mice and nonhuman primates showed no systemic cytotoxicity upon injection of recombinant TNF-related apoptosis inducing ligand (TRAIL) at doses that effectively suppressed solid tumors such as colon and mammary carcinomas. Although unwanted effects of some TRAIL preparations have been reported in normal cells, these data suggest that TRAIL could be a suitable approach in cancer therapy. However, several mechanisms of resistance to TRAIL-mediated apoptosis have been described in tumor cells such as lack of TRAIL apoptotic receptors, enhanced expression of TRAIL-decoy receptors, and expression of apoptosis inhibitors. In combination regimes, interferon-gamma (IFN-gamma) could provide a promising antitumor therapeutic approach as it has been described to enhance cellular susceptibility to apoptosis in a variety of tumor cells. The mechanism by which IFN-gamma promotes cell death seems to be via the regulation of the expression of different proteins involved in apoptosis. Altogether, these data suggest a combination strategy to selectively kill tumor cells that need to be further explored. PMID- 15110184 TI - Retinoids and TRAIL: two cooperating actors to fight against cancer. AB - Multiple studies performed in in vitro and in vivo settings have confirmed the cancer therapeutic and cancer preventive capacity of retinoids and rexinoids. These compounds mediate their actions through the retinoid and rexinoid receptors, respectively, which exist in multiple isoforms and form a plethora of distinct heterodimers. Despite their apparent anticancer potential, with one exception the molecular basis of this activity has remained largely elusive. The exception concerns acute promyelocytic leukemia (APL), the prototype of retinoic acid-dependent differentiation therapy, for which both the molecular nature of the disease and the mechanism of action of retinoids are well understood. However, retinoids and rexinoids are active beyond the borderlines of the well defined chromosomal translocation that gives rise to curable APL. In this context, particularly interesting is that retinoic acid induces a member of the tumor necrosis factor family, tumor necrosis factor-related apoptosis inducing ligand (TRAIL) or Apo2L. This ligand is exceptional in that it is capable of inducing apoptosis in cancer cells but not in normal cells. It is possible that this connection to the TRAIL signaling pathway contributes to the anti-tumor activity of retinoids and rexinoids. This review focuses on what is presently known about the regulation of cell life and death by the retinoid/rexinoid and TRAIL signaling pathways. PMID- 15110185 TI - Potential for TRAIL as a therapeutic agent in ovarian cancer. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is known to induce apoptosis, otherwise known as programmed cell death, in many malignant cells without any known detrimental effects to normal cells. These aspects of TRAIL indicate the potential of TRAIL as a therapeutic agent in cancer. Ovarian cancer remains the deadliest gynecologic malignancy and is the fourth leading cause of death due to cancer in women. However, it has been shown in studies that ovarian cancer cells are sensitive to TRAIL-induced cell death when treated with TRAIL alone or in combination with chemotherapeutic agents. TRAIL signals through two death receptors, TRAIL-R1 and TRAIL-R2, to induce apoptosis. TRAIL also binds to two other cell surface receptors, TRAIL-R3 and TRAIL-R4, which do not have intracellular death domains and therefore do not transmit the apoptotic signal upon ligation with TRAIL. It has been shown that a chemokine, interleukin-8 (IL 8), may play a role in ovarian tumor progression due to its elevated presence in the fluid surrounding ovarian cancer tissues. Possible roles for IL-8 in ovarian tumorigenesis include angiogenesis and metastasis. Because the mechanism of regulation for TRAIL-induced apoptosis needs to be clarified, the role of IL-8 in TRAIL-induced apoptosis of ovarian cancer cells was studied. Results showed that the presence of IL-8 regulates cell-surface expression of TRAIL receptors in ovarian cancer cell lines in vitro. There may be a role for the p38 mitogen activated protein kinase (MAPK) pathway in TRAIL-induced apoptosis of ovarian cancer cell. PMID- 15110186 TI - TRAIL and chemotherapeutic drugs in cancer therapy. AB - Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL/Apo2L) is a recently identified member of the TNF ligand family that selectively induces apoptosis in tumor cells in vitro and in vivo but not in most normal cells. Chemotherapeutic drugs induce apoptosis and the upregulation of death receptors or activation of intracellular signaling pathways of TRAIL. Numerous chemotherapeutic drugs have been shown to sensitize tumor cells to TRAIL-mediated apoptosis. Studies from our laboratory have also shown that TRAIL-resistant renal cell carcinoma, prostate gland cancer, and bladder cancer cells are sensitized by subtoxic concentrations of chemotherapeutic drugs including doxorubicin, epirubicin, pirarubicin, and cisplatin. TRAIL, particularly in combination with chemotherapeutic agents, is thus potentially promising in the treatment of cancer. This review addresses the putative role of TRAIL in cancer treatment and discusses the molecular basis of the synergistic effect of TRAIL and chemotherapeutic drugs. PMID- 15110187 TI - Additive effects of TRAIL and paclitaxel on cancer cells: implications for advances in cancer therapy. AB - In cancer therapy outgrowth of chemoresistant tumor cells is the most important factor that ultimately determines-apart from immediate adverse effects during treatment-the life span and prognosis of cancer patients. Despite many advances in cancer treatment and the integration of supportive medications, including new and better drugs for pain management, antiemesis, infection, and reconstitution of the hematopoietic system, both toxic effects and the development of resistance in response to the treatment remain a major problem. New treatment regimens have to be developed to target cancer more specifically using multiple cellular pathways. This will reduce toxic effects as well as the development of chemoresistance. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is the ligand for death receptors that belong to the TNF death receptor family. TRAIL triggers apoptosis in vitro in various cancer cell types. The antitumor drug, Paclitaxel (PA) was shown to increase the survival of patients with cancer. In in vitro experiments, PA also induces apoptosis in cancer cells. Together, PA and TRAIL lead to tumor regression in in vivo therapy and induce apoptosis through the interaction of TNF family death receptors, caspase activation, and?or cytochrome c release from mitochondria. PA and TRAIL complement each other using two distinct pathways that trigger apoptosis in addition to the anti-microtubule effect of PA. The combination of TRAIL and PA suppresses tumor growth that is otherwise resistant to treatment with either PA or TRAIL alone, by improving proapoptotic effects of the drugs. This observation support the use of the PA and TRAIL in future clinical trials. PMID- 15110188 TI - Regulation of sensitivity to TRAIL by the PTEN tumor suppressor. AB - The ability of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) to induce apoptosis preferentially in cancer cells is attractive for its development as a novel cancer therapeutic agent, but many cancer cell lines are resistant to TRAIL. While the molecular basis for TRAIL resistance is not always clear, a number of factors have been proposed to mediate TRAIL resistance, including decoy receptor, c-FLIP, nuclear factor (NF)-kappaB, and activation of antiapoptotic kinase signaling. Many growth factor receptors mediate their survival signals through the pathway involving recruitment and activation of phosphatidylinositol (PI) 3-kinase and the serine?threonine kinase Akt. The PTEN tumor suppressor is a phosphatase that dephosphorylates the phospholipids phosphorylated by PI-3 kinase, thereby opposing the action of PI 3-kinase, and acts as the primary negative regulator of the PI-3 kinase?Akt pathway in the cell. Loss of PTEN function occurs frequently in human tumors and leads to constitutive activation of Akt in cancer cells. Constitutively active Akt protects cells from TRAIL induced apoptosis in multiple tumor types. Growth factors such as epidermal growth factor or insulin-like growth factor-1 also inhibit TRAIL-induced apoptosis through the Akt pathway. Akt exerts its antiapoptotic function by its ability to phosphorylate many key components of the cellular apoptotic regulatory circuit, such as BAD, MDM2, FOXO Forkhead transcription factors, and PED?PEA-15 as well as by its role in activating NF-kappaB. Because PTEN loss is common in tumors, strategies to inactivate Akt may be necessary to overcome TRAIL resistance and make TRAIL-based therapy more effective. PMID- 15110189 TI - TRAIL and malignant glioma. AB - Encouragingly, some types of cancer can now be considered treatable, with patients reasonably expecting their disease to be cured. Chemotherapy and radiation therapy are effective against these cancers because they activate the so-called intrinsic apoptosis pathways within the cancer cells. Unfortunately currently available treatments are only effective against a subset of tumor types. In contrast, other cancers, such as malignant glioma, typically do not respond to currently available therapies. Some of this resistance can be attributed to these tumor cells failing to undergo apoptosis upon anticancer treatment. Recently, considerable research attention has focused on triggering apoptosis in chemotherapy- and radiation-therapy-resistant cancer cells via an alternative route-the "extrinsic" pathway, as a means of bypassing this block in apoptosis. Binding of members of the tumor necrosis factor-alpha (TNF-alpha) family of death ligands to their receptors on the cell surface triggers this pathway. Death ligands can kill some cancer cells that are resistant to the apoptotic pathway triggered by conventional anticancer treatments. Some death ligands, such as TNF-alpha and FasL, cause unacceptable toxicity to normal cells and are therefore not suitable anticancer agents. However another death ligand, TNF-related apoptosis-inducing ligand (TRAIL)/Apo-2L, and antibodies that emulate its actions, show greater promise as candidate anticancer drugs because they have negligible effects on normal cells. This review will discuss the ability of TRAIL to induce apoptosis in malignant glioma cells and the potential clinical applications of TRAIL-based agents for glioma treatment. PMID- 15110190 TI - Regulation of TRAIL-induced apoptosis by ectopic expression of antiapoptotic factors. AB - The discovery of an agent that selectively kills tumor cells and not normal cells is the dream of every cancer researcher. Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), first discovered in 1995, was heralded as a selective killer of tumor cells, and its potential is still thought to be high. Almost immediately, broad efforts were made to understand its activity at the molecular level. TRAIL has been shown to interact with the cell surface through five distinct receptors, named death receptor (DR) 4, DR5, decoy receptor (Dc)R1, DcR2, and osteoprotegrin. It activates nuclear factor (NF)-kappaB, c-Jun N terminal kinases, and apoptosis. The apoptotic signals are mediated through Fas associated death domain protein (FADD)-mediated recruitment of caspase-8 and caspase-3. Additionally, caspase-8 can cleave Bcl-2 homology domain 3 (BH3) interfering domain death agonist (Bid), and the cleaved Bid then causes the release of mitochondrial cytochrome c, leading to the activation of pro-caspase 9, which can then activate pro-caspase-3. TRAIL-induced apoptosis is negatively regulated by numerous cellular factors including decoy receptors, cellular FADD like interleukin 1 beta-converting enzyme (FLICE) interacting protein (cFLIP), cellular inhibitor of apoptosis protein (cIAP), X-linked IAP (XIAP), survivin, and NF-kappaB. Second mitochondria-derived activator of caspases (Smac)?direct IAP binding protein with low pI (DIABLO) mediates proapoptotic signals through inaction of IAP. How the TRAIL-induced apoptosis is downregulated by these factors is discussed in detail in this review. Whether TRAIL selectively kills tumor cells without harming normal cells is also discussed. PMID- 15110192 TI - Carbanion versus hydride transfer mechanisms in flavoprotein-catalyzed dehydrogenations. AB - The present understanding of the mechanisms by which flavoproteins oxidize amino acid or hydroxy acids to the respective imino or keto acids is reviewed. The observation that many of these enzymes catalyze the elimination of HBr or HCl from the appropriate beta-halogenated substrate was long considered evidence for a carbanion intermediate. Recent structural and mechanistic studies are not compatible with the intermediacy of carbanions in the reactions catalyzed by d amino acid oxidase and flavocytochrome b(2). In contrast, the data are most consistent with mechanisms involving direct hydride transfer. PMID- 15110193 TI - Electron transfer in natural and unnatural flavoporphyrins. AB - The development of chemical models for enzymes and their chemical and physical studies constitutes an important area of research from a scientific as well as an industrial point of view. Covalently linked flavin and porphyrin (flavoporphyrins) have attracted attention due to their applications as chemical models for flavoproteins and related enzymes. In this review, the literature has been surveyed to provide a comprehensive coverage of the synthetic methodology and characterization techniques of various types of synthetic flavoporphyrins. PMID- 15110194 TI - Convenient solid-phase synthesis of oligopeptides using pentacoordinated phosphoranes with amino acid residue as building blocks. AB - The reactive intermediates of pentacoordinated phosphoranes with amino acids (P(5)-AA) as building blocks, which were obtained by the reaction of O-phenylene phosphorochloridate with N,O-bis(trimethylsilyl)amino acids, were linked to a solid-phase support containing a hydroxymethyl polystyrene functional group. The first amino acid residue was coupled to the solid-phase support after washing the resin with organic solvent. Repeating the procedure led to oligopeptides linked on the resin. A series of free oligopeptides including tetra-Gly, di-Val, tri Val, di-Leu, di-Phe, and Phe-Leu were obtained after cleavage from solid-phase support. The structure of these oligopeptides were determined by IR, (1)H NMR, FAB-MS, and HPLC. PMID- 15110195 TI - Slow-binding inhibition of peptide deformylase by cyclic peptidomimetics as revealed by a new spectrophotometric assay. AB - A new spectrophotometric/fluorimetric assay for peptide deformylase (PDF) has been developed by coupling the PDF reaction with that of dipeptidyl peptidase I (DPPI) and using N-formyl-Met-Lys-AMC as substrate. Removal of the N-terminal formyl group by PDF renders the dipeptide an efficient substrate of DPPI, which subsequently removes the dipeptidyl units to release 7-amino-4-methylcoumarin as the chromophore/fluorophore. The PDF reaction is conveniently monitored on a UV Vis spectrophotometer or a fluorimeter in a continuous fashion. The utility of the assay was demonstrated by determining the catalytic activity of PDF and the inhibition constants of PDF inhibitors. These studies revealed the slow-binding behavior of a previously reported macrocyclic PDF inhibitor. This method offers several advantages over the existing PDF assays and should be particularly useful for screening PDF inhibitors in the continuous fashion. PMID- 15110196 TI - Myocardial perfusion grade and survival after percutaneous transluminal coronary angioplasty in patients with cardiogenic shock. AB - We sought to evaluate myocardial reperfusion and its prognostic value after percutaneous transluminal coronary angioplasty (PTCA) in patients admitted for cardiogenic shock. Lack of myocardial reperfusion despite restored coronary flow affects the survival of patients with acute myocardial infarction (AMI). Myocardial blush grade (MBG) is an angiographic measure of myocardial perfusion. We assessed MBG in 41 consecutive patients admitted to our department within 12 hours from the onset of AMI and in cardiogenic shock. PTCA was successful in 83% of patients. Thrombolysis In Mycardial Infarction (TIMI) grade 3 flow was demonstrated in 22 patients (53%). MBG 2/3 was found in 14 patients (34%); among them, 12 had TIMI 3 flow. Compared with patients with MBG 2/3, those with MBG 0/1 were older (71 +/- 11 vs 57 +/- 13 years, p = 0.001), had a higher prevalence of diabetes (48% vs 14%, p = 0.04) and hypertension (63% vs 29%, p = 0.04), showed a trend toward longer ischemic time (6.1 +/- 2.4 vs 4.9 +/- 1.1), and had larger enzymatic infarct size (peak creatine kinase 7,690 +/- 3,516 vs 5,500 +/- 2,977 IU/L). Mortality was higher in patients with MBG 0/1 both in the hospital (81% vs 14%, p <0.001) and at follow-up (81% vs 29%, p = 0.001). After adjustment by multivariate analysis, MBG 0/1 (odds ratio 16, p = 0.01) and age (odds ratio 3.8/10 years, p = 0.04) were correlated with in-hospital mortality. MBG 2/3 was achieved in a few patients in cardiogenic shock after AMI who were treated with PTCA; this was a strong predictor of in-hospital survival. Also, risk stratification after mechanical revascularization should include assessment of restoration of myocardial reperfusion. PMID- 15110197 TI - Usefulness and safety of percutaneous myocardial laser revascularization for refractory angina pectoris. AB - This prospective, double-blind, randomized, sham-controlled trial was designed to control for patient and investigator bias in assessing symptomatic improvement after percutaneous myocardial laser revascularization (PMLR) therapy. Eighty-two patients with stable angina pectoris (class III or IV) not amenable to conventional revascularization and with evidence of reversible ischemia, ejection fraction >/=25%, and myocardial wall thickness >/=8 mm were randomized to either PMLR with optimal medical therapy (n = 40) or to a sham procedure with optimal medical therapy (n = 42). With the exception of 1 laser technician, all patients, investigators, and assessors were blinded to treatment through the 12-month follow-up. The primary end point was restricted to Canadian Cardiovascular Society angina class improvement to limit the number of patients exposed to a sham procedure. Secondary assessments included medication usage, quality of life, exercise testing, ejection fraction, and hospitalizations. The incidence of serious adverse events, as determined by cardiac event-free survival at 12 months, was similar between groups. At 12 months, Canadian Cardiovascular Society angina scores improved by >/=2 classes in significantly more PMLR-treated patients than sham control patients (35% vs 14%, p = 0.04). Angina-specific quality-of-life measures were significantly higher in the PMLR group at each follow-up (p <0.05). Exercise and medication usage was similar between groups at 12 months. We conclude that PMLR therapy is reasonably safe and effective as symptomatic improvement in patients refractory to medical therapy, and that the clinical benefit is not attributable to placebo effect or investigator bias. PMID- 15110198 TI - Comparison of bivalirudin versus heparin during percutaneous coronary intervention (the Randomized Evaluation of PCI Linking Angiomax to Reduced Clinical Events [REPLACE]-1 trial). AB - To assess the efficacy of the direct thrombin inhibitor bivalirudin relative to heparin during contemporary coronary intervention, 1,056 patients who underwent elective or urgent revascularization were randomized in a large-scale pilot study to receive heparin (70 U/kg initial bolus) or bivalirudin (0.75 mg/kg bolus, 1.75 mg/kg/hour infusion during the procedure). All patients received aspirin; pretreatment with clopidogrel was encouraged, and glycoprotein (GP) IIb/IIIa blockade was at the physician's discretion. Stents were placed in 85% of patients; 72% received a GP IIb/IIIa inhibitor, and 56% were pretreated with clopidogrel. Activated clotting times were higher among patients randomized to bivalirudin than among those given heparin before device activation (median 359 vs 293 seconds, p <0.001). The composite efficacy end point of death, myocardial infarction, or repeat revascularization before hospital discharge or within 48 hours occurred in 5.6% and 6.9% of patients in the bivalirudin and heparin groups, respectively (p = 0.40). Major bleeding occurred in 2.1% versus 2.7% of patients randomized to bivalirudin or heparin, respectively (p = 0.52). This trial represents the largest prospective dataset of bivalirudin administered concomitantly with planned GP IIb/IIIa blockade and provides evidence of the safety and efficacy of this combined antithrombotic approach. PMID- 15110199 TI - Validation of viability assessment by electromechanical mapping by three dimensional reconstruction with dobutamine stress echocardiography in patients with coronary artery disease. AB - We evaluated the ability of electromechanical mapping (EMM) to discriminate between normal, viable, and nonviable (scarred) myocardium in patients with coronary artery disease versus dobutamine stress echocardiography (DSE) when the correspondence between the test and reference data sets is established via a common 3-dimensional reconstruction of the left ventricle. We studied 21 patients with coronary artery disease who underwent angiography, biplane ventriculography, and EMM within 1 month of DSE. A 3-dimensional left ventricular (LV) reconstruction was prepared from the ventriculogram and spatially aligned with EMM. EMM measurements of unipolar voltage, bipolar voltage, and local linear shortening were projected onto the three-dimensional left ventricle, averaged in each of 16 segments, and compared with DSE viability (normal, viable, scar) assessed at a core laboratory. All of the EMM measurements varied significantly (p <0.001) between the normal, viable, and scarred myocardium as assessed by DSE. Local linear shortening for normal, viable, and scarred segments was 10.4 +/- 6.5%, 7.8 +/- 5.6%, and 4.8 +/- 4.4%, respectively. In discriminating between these 3 groups, local linear shortening was more powerful than unipolar voltage or bipolar voltage (F = 20.765, F = 10.655, F = 4.795, respectively). Local linear shortening correlated best with viability, perhaps because it shares the same cognitive function as DSE. Three-dimensional analysis provides an anatomic framework that enables direct comparison of data from multiple imaging modalities rather than assuming segmental correspondence. Our results show that EMM provides significant on-line, diagnostic information on myocardial viability assessed by DSE on a segment-by-segment basis. PMID- 15110200 TI - Effect of acute myocardial infarction on the utility of fractional flow reserve for the physiologic assessment of the severity of coronary artery narrowing. AB - Fractional flow reserve (FFR) has been shown to be a useful physiologic index of coronary lesion severity in myocardial beds of patients without prior infarction and in those with remote infarction. Acute myocardial infarction (AMI) causes myocardial necrosis and microvascular stunning, embolization, and damage. Whether FFR remains a useful index of epicardial flow in the setting of recent myocardial infarction is not established. Cardiac risk factors, serum troponin I, angiographic minimal lumen diameter (MLD), percent diameter stenosis (DS), lesion length, vessel reference diameter, hyperemic central aortic pressure, hyperemic pressure distal to stenosis, and FFR were compared in 43 vessels subtending recent AMI beds to 25 control vessels, matched by lesion length and MLD, in patients without AMI. There were no differences in DS, MLD, lesion length, or reference diameter between AMI and non-AMI groups. Patients with AMI had mean troponin I levels of 91.8 +/- 162 ng/ml. Left ventricular ejection fraction was significantly lower in patients with than without AMI (55 +/- 9% vs 62 +/- 8%, p <0.05). There were no significant differences in hyperemic central aortic pressure (92 +/- 13 vs 99 +/- 15 mm Hg, p = NS), hyperemic pressure distal to the stenosis (62 +/- 17 vs 66 +/- 19 mm Hg, p = NS), or FFR (0.67 +/- 17 vs 0.68 +/- 17, p = NS) between recent AMI and non-AMI control patients. There was a significant correlation between DS and FFR for both patients with (p <0.001) and without (p = 0.003) infarctions. Thus, FFR and the relation between FFR and DS of lesions subtending AMI was not significantly different from FFR of angiographically matched lesions in patients without AMI. PMID- 15110201 TI - Spectrum of remodeling behavior observed with serial long-term (>/=12 months) follow-up intravascular ultrasound studies in left main coronary arteries. AB - Most intravascular ultrasound (IVUS) studies of arterial remodeling in native coronary arteries reported a remodeling index obtained at a single time point. We analyzed serial IVUS examinations, including the vessel cross-sectional area changes (remodeling behavior), of 60 hemodynamically nonstenotic left main lesions (baseline vs 18.4 +/- 9.4 months follow-up). Lumen reduction resulted from vessel reduction (sometimes despite plaque + media decrease), plaque + media increase (with or without vessel increase), or both. The percent annual changes in lumen area correlated strongly with changes in vessel (r = 0.84), but not with changes in plaque + media area. Plaques were classified as group A lesions, reflecting positive remodeling behavior (vessel changes >0), or group B lesions, reflecting negative (or intermediate) remodeling behavior (vessel changes <==0). Both groups did not differ significantly in demographics, laboratory data, and medications. Group A lesions (n = 40) more often showed plaque + media increase than group B lesions (32 of 40 [80%] vs 9 of 20 [45%]; p = 0.02). Group A lesions had, on average, mild annual lumen increase despite mild plaque + media increase, i.e, overcompensation of remodeling for plaque + media increase (vessel increase greater than plaque + media area increase, 19 of 40 [47%]). Conversely, group B lesions (n = 20) showed a significant lumen area reduction (-2.8 +/- 2.6 mm(2)/year) as a result of a decrease in vessel area only. Thus, serial long-term reduction of lumen size may result from vessel shrinkage (sometimes despite plaque decrease), plaque increase (with or without vessel increase), or both; overall, only the remodeling behavior has a significant relation to lumen changes. More than 30% of lesions show a negative remodeling behavior, which shows no relation to patient characteristics or initial plaque burden. PMID- 15110202 TI - Differences in heart rate turbulence between patients with coronary artery disease and patients with ventricular arrhythmias but structurally normal hearts. AB - The term "heart rate turbulence" (HRT) indicates the physiologic changes in the sinus cycle that follow a ventricular premature complex; impaired HRT denotes abnormalities in cardiac autonomic function. To investigate whether HRT is impaired in patients with stable coronary artery disease (CAD), we studied 29 patients with documented CAD and frequent (>/=30/hour) ventricular premature complexes on Holter monitoring and 31 patients with frequent ventricular arrhythmias but normal hearts (NH-VA). HRT and heart rate variability analyses were analyzed on 24-hour Holter recordings. HRT variables differed significantly between the 2 groups (turbulence onset -0.20 +/- 1.7% vs -0.67 +/- 2.2%, p = 0.00001; turbulence slope 2.83 +/- 1.9 vs 10.83 +/- 7.4 ms/RR, p = 0.0001 in patients with CAD and NH-VA, respectively). The difference was independent of a history of previous myocardial infarction, left ventricular function, and age. Top quartile turbulence onset values (>-0.26%) and bottom quartile turbulence slope values (<2.12 ms/RR) had similar predictive power in discriminating between patients with CAD and NH-VA (positive predictive value 86.7%, negative predictive value 64.4% for both). Among heart rate variables, bottom quartile SD of all RR intervals values (<96.3 ms) only had the same power of HRT variables in discriminating between patients with CAD and NH-VA. Thus, our data show that HRT variables are impaired in patients with CAD patients versus those with NH-VA, indicating abnormalities in the control of short-term cardiac autonomic mechanisms resulting in decreased vagal activity with likely predominant sympathetic activity. PMID- 15110203 TI - Meta-analysis of observational studies on fish intake and coronary heart disease. AB - Fish consumption has been associated with a lower risk of coronary heart disease (CHD) in some but not all studies. We conducted a meta-analysis of observational studies to determine if fish consumption is associated with lower fatal and total CHD. English language articles published before May 2003 were searched. In all, 19 observational studies (14 cohort and 5 case-control) in which there was a group that consumed fish on a regular basis and a comparison group that consumed little or no fish were included. With use of a standardized protocol and data extraction form, information on study design, sample size, participant characteristics, duration of follow-up, assessment of end points, and consumption of fish was abstracted. Using a random effects model, we pooled data from each study. Fish consumption versus little to no fish consumption was associated with a relative risk of 0.83 (95% confidence interval 0.76 to 0.90; p <0.005) for fatal CHD and a relative risk of 0.86 (95% confidence interval 0.81 to 0.92; p <0.005) for total CHD. The results indicate that fish consumption is associated with a significantly lower risk of fatal and total CHD. These findings suggest that fish consumption may be an important component of lifestyle modification for the prevention of CHD. PMID- 15110204 TI - Statin therapy is associated with lower mortality among patients with severe heart failure. AB - Experimental considerations suggest both potential harm and benefit from statin therapy in patients with severe heart failure. However, relations of statin therapy with clinical outcomes in severe heart failure are not well established. Using data from the Prospective Randomized Amlodipine Survival Evaluation (PRAISE) trial, we evaluated associations of statin therapy with total mortality among 1,153 patients with severe heart failure (ejection fraction <30% and New York Heart Association class IIIB or IV symptoms) of ischemic and nonischemic etiologies. Statin therapy was administered to 134 patients (12%) during the study period. Over a 1.3-year mean follow-up, there were 413 deaths (29 deaths/100 person-years). Adjusting for age, gender, diabetes, smoking, heart failure etiology, ejection fraction, and New York Heart Association class, statin therapy was associated with a 62% lower risk of death (hazard ratio 0.38, 95% confidence interval 0.23 to 0.65), or 1 fewer death/5 patients taking statin therapy for 1 year. This association was not greatly altered by additional adjustment for a variety of other patient characteristics, including serum cholesterol levels. After propensity score analyses, statin therapy was still associated with a 48% lower risk of death (hazard ratio 0.52, 95% confidence interval 0.30 to 0.89). Although this observational study does not prove causality, further investigation of potential benefits of statins in patients with severe heart failure appears warranted. PMID- 15110205 TI - Comparative accuracy of B-type natriuretic peptide and tissue Doppler echocardiography in the diagnosis of congestive heart failure. AB - B-type natriuretic peptide (BNP) and early diastolic transmitral velocity/tissue Doppler mitral annular velocity (E/Ea) both estimate left ventricular filling pressure, but have not been compared in the diagnosis of congestive heart failure (CHF). One hundred twenty-two hospital inpatients with suspected CHF underwent simultaneous clinical examination, BNP measurement, and comprehensive echo Doppler examination. The accuracy of BNP and echocardiography was compared with the Framingham criteria diagnosis of CHF. Seventy patients (57%) had clinical CHF, whereas 52 (43%) did not. In all patients, the optimal BNP cutoff was >250 pg/ml (sensitivity 86%, specificity 77%). E/Ea >15 had 83% sensitivity and 82% specificity, whereas comprehensive echo-Doppler had 95% sensitivity and 88% specificity for CHF. In patients with normal ejection fraction, the optimal BNP cutoff was >150 pg/ml (sensitivity 79%, specificity 85%). E/Ea >15 had 79% sensitivity and 93% specificity, whereas comprehensive echo-Doppler had 85% sensitivity and 96% specificity for CHF. In patients with reduced ejection fraction, the optimal BNP cutoff was >300 pg/ml (sensitivity 88%, specificity 60%). E/Ea >15 had 92% sensitivity and 72% specificity, whereas comprehensive echo-Doppler had 96% sensitivity and 80% specificity (p = 0.08 compared with BNP) for CHF. Overall, BNP and E/Ea have similar diagnostic accuracy for CHF in this patient population. In patients with reduced ejection fraction, comprehensive echo-Doppler trended toward higher specificity than BNP for clinical CHF. PMID- 15110206 TI - T-wave axis deviation as an independent predictor of mortality in chronic Chagas' disease. AB - The T-wave axis shift has been reported to represent a general marker of ventricular repolarization abnormalities and a potential indicator of increased risk for cardiovascular mortality. We assessed the prognostic importance of the T wave axis deviation for mortality rate in patients with chronic Chagas' disease. In a long-term follow-up prospective study, 738 adult outpatients in the chronic phase of Chagas' disease were enrolled. The frontal plane T-wave axis was estimated from 12-lead electrocardiograms obtained on admission and categorized as normal (15 degrees to 75 degrees ), borderline (75 degrees to 105 degrees or 15 degrees to -15 degrees ), and abnormal (>105 degrees or < -15 degrees ). Clinical and radiologic data, 2-dimensional echocardiographic data, and other electrocardiographic data were also recorded. Primary end points were all-cause, those related to Chagas' disease, and sudden cardiac deaths. Statistical analyses included Kaplan-Meier estimation of survival curves and multivariate Cox's proportional hazards models. During a follow-up of 58 +/- 39 months, 62 patients died, 54 from causes related to Chagas' disease and 40 due to sudden cardiac death. Kaplan-Meier survival curves showed that the 3 categories of T axis had significantly different prognoses. Multivariate Cox's survival analysis demonstrated that an abnormal T axis increases the risk of death threefold and sudden death nearly sixfold after adjustment for other covariates, including left ventricular systolic function and other electrocardiographic abnormalities. Borderline T-wave axis also indicated a worse prognosis, particularly in the subgroup of patients with abnormal baseline electrocardiograms. These results indicate that T-wave axis deviation is an easily quantified, strong, and independent mortality risk predictor in patients with chronic Chagas' disease. PMID- 15110207 TI - Long-term outcome and quality of life in adult patients after the Fontan operation. AB - The first successful Fontan operation was performed in 1971, and this first cohort of Fontan patients is reaching adulthood with unclear outcome of this palliative procedure. We studied the mortality, morbidity, and quality of life in our adult Fontan patients. We examined all patients (n = 36) who underwent a Fontan procedure and were being seen in an adult outpatient clinic by using electrocardiography, exercise testing, and echocardiography. Quality of life was assessed by the Short Form 36 questionnaire. The mean follow-up period was 15 years (range 0 to 23). Of the initial 36 patients, 10 died (28%) at a mean of 10 years (range 0 to 21) after the Fontan operation and 1 patient underwent cardiac transplantation. Reoperations were performed in 21 patients (58%), and the most common reason was revision of the Fontan connection. Sustained supraventricular tachycardia was observed in 20 patients (56%) with an increased incidence of arrhythmias with longer follow-up. Thromboembolic events were detected in 9 patients (25%), 5 of whom had adequate anticoagulant levels at the time of event. The thromboembolic event was fatal for 3 patients. A total of 195 hospital admissions (mean 3.8 +/- 2.7, range 1 to 13) was recorded. Quality-of-life assessment showed physical functioning, mental health, and general health perception to be significantly lower for Fontan patients than for the normal Dutch population. Thus, we found high mortality and very high morbidity in adult patients after the Fontan operation. In particular, reoperations, arrhythmias, and thromboembolic events compromised quality of life. PMID- 15110208 TI - Coronary artery calcium volume scores on electron beam tomography in 12,936 asymptomatic adults. AB - We developed age- and gender-specific normative tables of calcium volume scores by using data from 12,936 asymptomatic patients who underwent electron beam tomographic scanning and compared the volume with the Agatston scores obtained in the same subjects. The 2 scores increased as the number of atherosclerotic risk factors increased. The volume scores were statistically smaller than the Agatston scores at the upper quartile level. PMID- 15110209 TI - Relation of coronary calcium scores by electron beam tomography to obstructive disease in 2,115 symptomatic patients. AB - This angiographically correlated study reports on, for the first time, age- and gender-based distribution of the volumetric calcium score in a large group of patients with suspected coronary artery disease. Volumetric calcium data predicted significant coronary artery disease (>/=50% lumen diameter stenosis) as well as the traditional Agatston score. Exclusion of any calcium was highly accurate in ruling out obstructive disease in symptomatic subjects >/=50 years of age. PMID- 15110210 TI - Interscan variability of coronary artery calcium quantification using an electrocardiographically pulsed spiral computed tomographic protocol. AB - The use of an electrocardiographically pulsed spiral computed tomographic protocol significantly reduced the radiation dose to patients who underwent coronary calcium screening. For Agatston scores <10, the interscan variability of such a protocol was significantly lower than that for a sequential acquisition protocol. At higher Agatston scores, the 2 protocols had similar variability characteristics. PMID- 15110211 TI - Left atrium remodeling after acute myocardial infarction (results of the GISSI-3 Echo Substudy). AB - To evaluate the existence, timing, and determinants of post-infarction left atrial remodeling, we studied a subgroup of 514 patients from the Third Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto Miocardico Echo Substudy who underwent 4 serial 2-dimensional echocardiograms up to 6 months after acute myocardial infarction. This study is the first to demonstrate, in a large series of patients, the existence of early and late left atrial remodeling after low risk acute myocardial infarction and the relation of left atrial remodeling to left ventricular remodeling. PMID- 15110212 TI - Usefulness of myocardial contrast echocardiography derived coronary flow reserve to accurately determine severity of left anterior descending coronary artery stenosis. AB - Thirty-five patients underwent quantitative low-power myocardial contrast echocardiography to evaluate the severity of left anterior descending coronary artery stenosis. Coronary flow reserve and myocardial blood flow were able to accurately differentiate among angiographically derived grades of coronary stenosis. PMID- 15110213 TI - Comparison of interventional versus conservative treatment of isolated ostial lesions of coronary diagonal branch arteries. AB - This study compares percutaneous coronary intervention of isolated ostial stenosis of diagonal branches with a luminal diameter >/=2.0 mm with medical treatment with regard to cardiac events during hospitalization and follow-up. Medical treatment is an alternative to percutaneous intervention without a greater incidence of death or myocardial infarction at 12-month follow-up. Interestingly, patients with isolated ostial stenosis of diagonal branches who were treated interventionally showed a significantly greater probability of rehospitalization for severe angina, recatheterization, and reintervention compared with medically treated patients. PMID- 15110214 TI - Relation of coronary artery ectasia to diabetes mellitus. AB - An increased prevalence of coronary artery ectasia (CAE) and a low frequency of diabetes mellitus have been reported in patients with abdominal aortic aneurysm (AAA). The prevalence of diabetes was studied in 190 patients with CAE in comparison with 341 age- and gender-matched patients with coronary artery disease alone. Diabetes mellitus was found to be independently but inversely associated with CAE (relative risk 0.603, 95% confidence interval 0.375 to 0.960, p = 0.037), thus resembling the relation between diabetes and AAA. PMID- 15110215 TI - Effects of race on lipid-lowering management in hospitalized patients with coronary heart disease. AB - The objective of this study was to determine the association between patient characteristics, specifically race, and the appropriate management of lipid reducing therapy in patients with coronary heart disease (CHD) discharged from the hospital. Two hundred fifty-eight consecutive patients with diagnoses suggestive of CHD were identified in a large, inner-city university teaching hospital serving a predominantly indigent African American population. The outcome measure, suboptimal lipid management, evaluated the intensification of lipid-reducing therapy when indicated using the National Cholesterol Education Program guidelines for the low-density lipoprotein cholesterol goal. The overall rate of suboptimal lipid management was 48%. Black patients with CHD were significantly more likely to have suboptimal lipid management than nonblack patients by the time of hospital discharge (52.3% vs 16.7%, p = 0.021). The disparity in the clinical management of black patients with CHD may help explain the differential in health outcomes seen between black and white patients. PMID- 15110216 TI - Impact of insulin-requiring diabetes mellitus on effectiveness of reperfusion and outcome of patients undergoing primary percutaneous coronary intervention for acute myocardial infarction. AB - The relation between diabetes mellitus (DM) and outcome was assessed in a series of 1,061 patients with acute myocardial infarction (AMI) who underwent primary percutaneous coronary intervention (PCI). The efficacy of reperfusion was assessed by ST-segment resolution analysis. Of 1,061 patients, 166 had DM (15.6%), and 84 had insulin-requiring DM (51% of DM patients). The 6-month mortality rate was 26% in insulin-requiring DM patients, 7% in non-DM patients, and 4% in non-insulin-requiring DM patients (p <0.001). The early ST-segment resolution rate was lower in insulin-requiring DM patients (52%) compared with the other DM patients (78%) and non-DM patients (76%; p <0.001). Multivariate analysis showed insulin-requiring DM to be independently related to the risk for death (hazard ratio 1.94, 95% confidence interval 1.17 to 3.22, p = 0.009). Insulin-requiring DM is a strong predictor of mortality in patients who undergo PCI for AMI, and this relation may be explained by a less effective myocardial reperfusion despite the mechanical restoration of normal epicardial flow in most patients. PMID- 15110217 TI - Early recurrence of arrhythmia in patients taking amiodarone or class 1C agents for treatment of atrial fibrillation or atrial flutter. AB - Amiodarone use for the prevention of recurrent atrial fibrillation or atrial flutter requires drug loading over a period of several days to weeks, whereas class 1C agents do not. Three hundred thirty-nine patients were evaluated during drug loading with amiodarone or class 1C agents, and it was found that recurrent arrhythmia was common, usually not persistent, and frequently asymptomatic. Recurrent arrhythmia was not more common with amiodarone. PMID- 15110218 TI - Relation of an exaggerated rise in white blood cells after coronary bypass or cardiac valve surgery to development of atrial fibrillation postoperatively. AB - This study investigated the correlation between an elevated white blood cell (WBC) count as a marker of inflammation and the development of atrial fibrillation (AF) after cardiac surgery. WBC counts were prospectively followed in 181 consecutive patients who underwent coronary bypass or cardiac valve surgery to determine if a baseline or postoperative WBC count elevation is an independent predictor of postoperative AF. PMID- 15110219 TI - Usefulness of echocardiographic tissue synchronization imaging to predict acute response to cardiac resynchronization therapy. AB - Echocardiographic tissue synchronization imaging (TSI) consists of color-coding time-to-peak tissue Doppler velocities. This study of 29 patients who underwent cardiac resynchronization therapy (CRT) demonstrated that differences in baseline time-to-speak velocities of opposing ventricular walls by TSI were greater in 15 patients, with an acute hemodynamic improvement. A >/=65 ms delay from the anterior septum to the posterior wall using the apical long-axis view had 87% sensitivity and 100% specificity for predicting an acute response. Although a subgroup without acute improvement had later decreases in end-systolic volume, suggesting that acute response underestimates long-term effects, TSI has potential to assist in guiding CRT. PMID- 15110220 TI - Relation between QT duration and mortality in an elderly Japanese population. AB - The effects of prolonged QTc intervals on mortality were investigated in about 3,500 elderly Japanese patients followed for approximately 8.8 years. Prolonged QTc was found to be a marker for risk for all-cause mortality and mortality from heart disease or from coronary heart disease (CHD) after adjusting for other CHD risk factors. Even in Japanese subjects, who have a lower coronary heart disease rate than that of Caucasians, the careful observation of subjects with a prolonged QTc is believed to be necessary. PMID- 15110221 TI - Long-term prognosis of early and late prosthetic valve endocarditis. AB - A prospective series of 78 cases of prosthetic valve endocarditis (PVE) was studied (35 cases of early PVE and 43 cases of late PVE). The in-hospital mortality rate was significantly higher in patients with early PVE (31% vs 9%, p <0.01) because the onset of heart failure was more common in these patients (55% vs 37%, p <0.05). However, event-free survival at 4 years in survivors to the active phase was not different (74% and 82%, respectively). PMID- 15110222 TI - Cardiopulmonary exercise testing before and one year after mitral valve repair for severe mitral regurgitation. AB - For the evaluation of efficacy of cardiopulmonary exercise testing, we compared New York Heart Association functional class with peak oxygen consumption rate (VO(2)peak) in 31 patients with severe mitral regurgitation who underwent mitral valve repair surgery. One year later, the VO(2)peak values did not show significant improvement; however, the patients who had more than a mild degree of residual mitral regurgitation (n = 14) after 1 year of surgery had a VO(2)peak value that was significantly decreased (from 22.7 +/- 6.4 to 21.0 +/- 6.3 ml/kg/min, p = 0.04). Patients with a higher preoperative VO(2)peak value (>/=18.5 ml/kg/min) had a significantly better New York Heart Association functional class 1 year after surgery than patients with a lower VO(2)peak value (<18.5 ml/kg/min, p = 0.03). PMID- 15110223 TI - Familial clustering of autoimmune diseases in patients with dilated cardiomyopathy. AB - In this study, we examined the hypothesis that dilated cardiomyopathy (DCM) shares genetic risk factors with other diseases of presumed autoimmune etiology, and, therefore, the same multiple genes in combination with environmental factors lead to numerous different autoimmune diseases. In accordance with this hypothesis, we showed an increased prevalence of autoimmune diseases in first degree relatives of patients with DCM. Also, T-cell activation, as reflected in high levels of the soluble interleukin-2 receptor, appears to identify patients with DCM with a clustering of autoimmune diseases. PMID- 15110224 TI - Long-term follow-up of implantable cardioverter defibrillator therapy for hypertrophic cardiomyopathy. AB - In this study, high-risk patients with hypertrophic cardiomyopathy were treated with an implantable defibrillator. Long-term follow-up analysis showed the efficacy of implantable defibrillator therapy, with patients having an appropriate device intervention at a rate of 11%/year, thereby preventing sudden death in an Australian population. PMID- 15110225 TI - Regional differences in rates of diagnosis and mortality of pulmonary thromboembolism. AB - The rates of diagnosis of deep venous thrombosis, pulmonary embolism (PE), and venous thromboembolism (PE and/or deep venous thrombosis) and the mortality rate of PE in hospitalized patients were evaluated in 4 regions of the United States. Data are from the National Hospital Discharge Survey and the United States Bureau of the Census. The western region of the United States showed a lower rate of diagnosis of deep venous thrombosis and venous thromboembolism and a lower mortality rate of PE than any other region. PMID- 15110226 TI - Estimated case fatality rate of pulmonary embolism, 1979 to 1998. AB - Trends in the estimated case fatality rate of pulmonary embolism (PE) in the United States were obtained from the Bureau of the Census Compressed Mortality File and the National Hospital Discharge Survey. The estimated case fatality rate of PE increased from 1979 to 1989, then decreased through 1998. It increased exponentially with age and was higher in African-American than Caucasian patients (rate ratio 1.42; 95% confidence interval 1.40 to 1.44). The declining population mortality rate of PE from 1979 to 1989 appears to be due to a decreased incidence of PE, reflective of improved prophylaxis, whereas the continued declining population mortality rate of PE through 1998 reflects a decreased case fatality rate due to earlier and better management. PMID- 15110227 TI - Anti-inflammatory and antioxidant actions of statins may favorably affect atrial remodeling in atrial fibrillation. PMID- 15110228 TI - Lymphoproliferative disorders in Sjogren's syndrome. AB - Sjogren's syndrome (SS) is a chronic organ-specific autoimmune disease characterized by lymphocytic infiltration into the salivary and lacrimal glands. About half of primary SS patients develop systemic disorders. Primary SS can be divided into three stages according to the extent of organ damage and the course of the disease. In stage I, (approx. 45% of cases), patients have only sicca syndrome and do not experience any systemic involvement, even after 10 years. In stage II (approx. 50% of cases), patients experience lymphocytic organ damage, which may involve the pulmonary, renal, hepatic, hematologic, and/or dermatologic systems, among others. Finally, in stage III (approx. 5% of cases), patients develop malignant lymphomas. Lymphomas in salivary glands are thought to arise from lymphoepithelial lesions in which there are close interactions among epithelial cells, T cells, and B cells. The B cells in the lesions become activated through the interaction between CD40L and CD40. The progression from polyclonal lymphoproliferation to monoclonal lymphoproliferation, to mucosa associated lymphoid tissue (MALT) lymphoma, and finally to high-grade malignant lymphoma is regarded as a multi-step process. Antigenic activation of B cells, together with oncogenic events, including p53 inactivation and bcl-2 activation, may play important roles in B cell monoclonal proliferation and malignant transformation. The rheumatoid factor clone is regarded as a candidate B cell clone that undergoes transformation. PMID- 15110229 TI - Dendritic cells and autoimmunity. AB - Dendritic cells (DC) are professional antigen-presenting cells that are specialized in the uptake of antigens and their transport from peripheral tissues to the lymphoid organs. Because of their capacity to stimulate naive T cells, DC have a central role in the initiation of primary immune responses and are considered promising tools and targets for immunotherapy. Emerging data suggest a role for DC in initiating autoimmune attacks. Direct analysis of DC phenotypes and DC-T-cell interactions in rodent and human autoimmune diseases should shed light on how pathogenesis occurs, and suggest novel avenues of treatment aimed at alleviating deviant DC function. PMID- 15110230 TI - P-glycoprotein in autoimmune diseases. AB - Multidrug resistance-1 (MDR-1) is characterized by overfunction of P-glycoprotein (P-gp), a pump molecule that decreases intracellular drug concentration by effluxing them from the intracellular space. Broad ranges of structurally unrelated compounds are transported by P-gp, including antineoplastic agents, HIV protease inhibitors, prednisone, gold salts, methotrexate, colchicine as well as several antibiotics. In contrast, many other compounds such as calcium channel blockers (verapamil) and immunosupressors (cyclosporine-A) are able to inhibit P gp function. The P-gp role in therapeutic failures has been extensively studied in cancer; however, there is little information regarding MDR-1 phenotype in autoimmune disorders. It has been reported that an increased number of lymphocytes are able to extrude P-gp substrates in rheumatoid arthritis, immune thrombocytopenic purpura and systemic lupus erythematosus, the patients with poor response to treatment being the ones that exhibit the highest values. This may be due, at least in part, to a simultaneous long-term usage of several drugs that induce P-gp function. Since abnormally activated cell compartments characterize autoimmune diseases, it is possible that those cells are the ones that exhibit drug resistance. The study of drug resistance mechanisms in autoimmunity may be helpful for the optimization of the current therapeutic schemes through their combination with low doses of P-gp inhibitors. PMID- 15110231 TI - Synovial fluid estrogens in rheumatoid arthritis. AB - Experimental and clinical evidence suggest that immune reactivity is modulated by gender. Immune reactivity is greater in females than in males and lymphocytes and monocytes from female subjects shows higher antigen presenting activity and mitogenic responses. Steroid hormones can be converted along defined pathways to downstream hormones in the periphery. The conversion of dehydroepiandrosterone (DHEA) in target macrophages leads to an increase of downstream effector hormones (including estrogens), which may be an important factor for local immunomodulation at least in RA synovitis. The presence in the RA synovial fluids (SF) of an altered sex hormone balance resulting in lower immunosuppressive androgens and higher immunoenhancing estrogens, might determine a favorable condition for the development of the immuno-mediated RA synovitis and synovial hyperplasia. The increased estrogen concentration observed in RA SF of both sexes are characterized by the hydroxylated forms, in particular 16alpha hydroxyestrone, that is a mitogenic and proliferative endogenous hormone. In contrast to 16alpha-hydroxylated estrogens, the 2-hydroxylated forms inhibit growth promoting effects of E2 and were found low in RA SF. Therefore, dose related conversion to pro- or anti-inflammatory downstream metabolites of estrogens might support the dual role of estrogens (pro or anti-inflammatory) for example during estrogen replacement therapy, depending on local concentration (i.e. SF in RA) of 16alpha-hydroxyestrone or 2-hydroxyestrogens. PMID- 15110232 TI - Oxidative stress and nutritional prevention in autoimmune rheumatic diseases. AB - The hypothesis that oxidative stress favours flogistic and immune processes inducing autoimmune rheumatic diseases (ARDs) and their complications is still under discussion. In this review we take into consideration both the aetiopathological role of the diet in such diseases and the possible efficacy of dietary supports as adjuvants for the usual specific therapies. Moreover, we shall examine the hypothetical pathophysiological role of oxidative stress on ARDs and their complications, the methods for its evaluation and the possibility of intervening on oxidative pathways by means of nutritional modulation. It is possible that in the future we will be able to control connective pathology by associating an immuno-modulating therapy ('re-educating') with natural products having an anti-oxidant activity to current immunosuppressive treatment (which has potentially toxic effects). PMID- 15110233 TI - Autoimmune hepatitis: evolving concepts. AB - The liver is continuously exposed to a large antigenic load that includes pathogens, toxins, tumor cells and dietary antigens. A loss of tolerance against its own antigens may result in autoimmune hepatitis (AIH). The current paradigm holds that the disease is the result of self-perpetuating autoimmune process triggered by yet unknown factors (infections, chemicals, drugs) in a genetically susceptible host. To date, several putative hepatocellular surface antigens have been identified: P450-IID6 (recognized by the anti-LKM-1 autoantibodies) a membrane bound asialoglycoprotein receptor (a liver-specific membrane protein), a cytosolic UGA-suppressor tRNA associated protein (recognized by anti-SMA and anti LP antibodies) and argininosuccinate lysate and formiminotransferase cyclodeaminase (recognized by ant-LC1 antibodies). In contrast to other chronic hepatitides patients with AIH display significant T cell hypereactivity to autologous liver antigens. Tissue injury seems to be mediated by CD4+ or CD8+ T cells and/or by antibody-dependent cell mediated cytotoxicity. PMID- 15110234 TI - CD4+ CD8+ double positive (DP) T cells in health and disease. AB - The expression of CD4 and CD8alphabeta co-receptors on mature T cells is generally considered to be mutually exclusive and reflects subset-related, specific functions (helper vs. cytolytic) and differences in major histocompatibility complex-restriction for antigen recognition. However, double positive (DP) T cells expressing both CD4 and CD8 have been described in several pathological conditions as well as in normal individuals. DP T cells represent a heterogeneous population. Strong evidence indicates that in vivo terminally differentiated effector CD4 may acquire the alpha-chain of CD8. Reciprocally, in vitro activation of CD8+ T cells results in the expression of low levels of CD4 that may mediate HIV entry and responses to chemotactic cytokines. Particularly intriguing, a subset of DP T cells expressing high levels of both CD4 and CD8alphabeta heterodimer (CD4(hi)CD8(hi)), has been identified in autoimmune and chronic inflammatory disorders. While no definitive proof exists, it could be speculated that CD4(hi)CD8(hi) T cells may be endowed with auto-reactivity due to faulty thymic selection. PMID- 15110235 TI - Inflammation and chronic heart failure-potential therapeutic role of intravenous immunoglobulin. AB - Accumulating evidence indicates that inflammatory cytokines play a pathogenic role in congestive heart failure (CHF) by influencing heart contractility, inducing hypertrophy and promoting apoptosis or fibrosis, contributing to the continuous myocardial remodelling process. Traditional cardiovascular drugs seem to have little influence on the overall cytokine network, and immunomodulatory therapy have emerged as a possible new treatment modality in CHF. Several animal studies have suggested that modulation of inflammatory cytokines may improve cardiac performance, and we have recently demonstrated that intravenous immunoglobulin enhances left ventricular ejection fraction in CHF patients, significantly correlated with anti-inflammatory effects of such therapy. While not necessarily the drugs of choice, these studies suggest a potential for immunomodulatory therapy in CHF in addition to optimal cardiovascular treatment regimens. Further research will have to more precisely identify the most important actors in the immunopathogenesis of CHF in order to develop more specific immunomodulating agents in this disorder. Although these shortcomings, we believe that the 'cytokine era' in cardiovascular research might lead to quite new treatment modalities in CHF patients potentially leading to reduced morbidity and mortality in these patients. PMID- 15110236 TI - Antiphospholipid antibodies: a major advance in the management of recurrent abortion. AB - Recurrent pregnancy loss is a vexing problem facing many couples and doctors. Repetitive losses affect 2-5% of couples and specific causes are identified in only approximately 50% of patients. Many changes have occurred in this field over the past 20 years and the technology available for the workup of patients has improved. However, the diagnostic yield and pregnancy outcomes of patients with recurrent abortion have not improved significantly over the last two decades. In this regard, the identification of aPL as a new and treatable cause of recurrent abortion represents the major advance in the subject over the past 20 years. PMID- 15110237 TI - 5th International symposium on IVIG: IVIG in the third millenium, 25-27 September 2003, Interlaken, Switzerland. PMID- 15110238 TI - Proceedings of the XVIIth International Symposium on Bioelectrochemistry and Bioenergetics. Florence, Italy, 19-24 June 2003. PMID- 15110239 TI - Self-assembled monolayers of organosulphur molecules bearing calix[4]arene moieties. AB - We investigate the self-assembly of modified calix[4]arene on gold surfaces. Calix[4]arene was modified through a reaction sequence which led to assembling of the crown-5 moiety and to the insertion of two thioether groups into the starting molecule. The so-obtained calix[4]arene-crown-5 bis(7-thiatridecyloxy) (hereafter called calix[4]arene) was in the stable 1,3-alternate conformation. The calix[4]arene/gold interface was investigated by means of spectroscopic ellipsometry (SE), scanning tunneling microscopy (STM) and cyclic voltammetry (CV). SE data indicate a layer thickness compatible with the formation of a monomolecular layer. This result is confirmed by STM imaging which shows the formation of a high density of small pits, one gold layer deep, a typical feature of self-assembled organosulphur monolayers on gold. CV measurements performed in presence of the [Ru(NH(3))(6)(2+/3+)] redox couple indicate a passivation of the metal electrode, resulting in a reduction of the redox current, after the layer deposition. CV has also been used to investigate the selectivity properties of calix[4]arene-covered gold electrodes by measuring the redox current decrease in the presence of different salt solutions. It is found that calix[4]arene-covered electrodes are able to complex K(+) and Ba(2+), while no complexation is observed in the case of Li(+), Na(+), Cs(+), Mg(2+) and Ca(2+). PMID- 15110240 TI - Immobilization of a fluorescent dye in Langmuir-Blodgett films. AB - We immobilized a hemicyanine dye, Di-8-ANEPPS, in ordered thin films of an organic matrix, dihexadecyl phosphate (DHP), and we transferred the mixed monolayers onto solid support by the Langmuir-Blodgett technique. We used gold and quartz slides and indium tin oxide (ITO) evaporated on glass slides as substrates. The multilayers formation was confirmed by ellipsometric and contact angle measurements. The optical response of the nanostructures was investigated collecting UV-Vis absorption and fluorescence emission intensity profiles. PMID- 15110241 TI - Langmuir-Blodgett films incorporating redox mediators for molecular recognition of NADH. AB - We focused our studies on the fabrication of nano-organized systems incorporating redox mediators for NADH. We fabricated gold-modified electrodes of phospholipid layers containing a redox mediator for NADH by means of the Langmuir-Blodgett (LB) technique. We chose tetramethylbenzidine (TMB) which is known to decrease the very large overvoltage involved in the direct oxidation of NADH. The electrochemical characterisation of the films allowed to determine the presence and the electrochemical activity of the mediator in the LB layers. The response of the sensing layer in solutions containing NADH was investigated by means of cyclic voltammetry and fluorescence spectroscopy. We observed that NADH has a strong affinity with the film, and it interacts with the TMB-containing phospholipid layers. In fact, we found that TMB molecules mediate the NADH oxidation reaction, even if the surrounding matrix relatively hinders their mobility. PMID- 15110242 TI - Cyclic voltammetric study of the redox system of glutathione using the disulfide bond reductant tris(2-carboxyethyl)phosphine. AB - The stabilization of the reduction state of proteins and peptides is very important for the monitoring of protein-protein, protein-DNA and protein xenobiotic interactions. The reductive state of protein or peptide is characterized by the reactive sulfhydryl group. Glutathione in the reduced (GSH) and oxidized (GSSG) forms was studied by cyclic voltammetry. Tris(2 carboxyethyl)phosphine (TCEP) as the disulfide bond reductant and/or hydrogen peroxide as the sulfhydryl group oxidant were used. Cyclic voltammetry measurements, following the redox state of glutathione, were performed on a hanging mercury drop electrode (HMDE) in borate buffer (pH 9.2). It was shown that in aqueous solutions TCEP was able to reduce disulfide groups smoothly and quantitatively. The TCEP response at -0.25 V vs. Ag/AgCl/3 M KCl did not disturb the signals of the thiol/disulfide redox couple. The origin of cathodic and anodic signals of GSH (at -0.44 and -0.37 V) and GSSG (at -0.69 and -0.40 V) glutathione forms is discussed. It was shown that the application of TCEP to the conservation of sulfhydryl groups in peptides and proteins can be useful instrument for the study of peptides and proteins redox behavior. PMID- 15110243 TI - Electroanalytical determination of d(GCGAAGC) hairpin. AB - Hairpins (or hairpin-like structures) may play a major role in expansion events of triplet repeat expansion diseases (X syndrome, Huntington's disease, Friedreich's ataxia). The d(GCGAAGC) fragment has been found in the replication origins of phage phiX 174 and herpes simplex virus, in a promoter region of an Escherichia coli heat-shock gene, and in rRNA genes. The paper deals with the application of electrochemical methods to the determination of the DNA heptamer d(GCGAAGC) which forms very stable hairpin structure in aqueous solutions. On mercury electrodes, this hairpin provides voltammetric reduction signals of adenine and cytosine, and oxidation signals of guanine. Both signals have been studied by cyclic voltammetry (CV), linear sweep voltammetry (LSV), and elimination voltammetry with linear scan (EVLS) in dependence on pH, accumulation time, scan rate, and loop sequences. The EVLS in combination with the adsorptive stripping was employed to the determination of the detection limit (LD) of this mini-hairpin (2 nM). Multidimensional voltammetric data were worked up by Fourier Transform (FT) and for the first coefficient a confidence ellipse was calculated in order to drop out some outlier data. The same method was used also for detection limit determinations. The values of LD obtained by two approaches were compared. PMID- 15110244 TI - Square wave and elimination voltammetric analysis of azidothymidine in the presence of oligonucleotides and chromosomal DNA. AB - Azidothymidine (AZT, 3'-azido-3'-deoxythymidine, Zidovudine, Retrovir) is an approved and widely used antiretroviral drug for the treatment of human immunodeficiency virus (HIV) infection. Dynamic electrochemical methods have been employed for the fast and inexpensive determination of this drug in natural samples. The electrochemical signal of AZT, resulting from the reduction of azido group, was studied by square wave voltammetry (SWV), linear sweep voltammetry (LSV) and elimination voltammetry with linear scan (EVLS) using a hanging mercury drop electrode (HMDE). This paper explores the possibility of determining AZT in the presence of native (dsDNA) or denatured calf thymus DNA (ssDNA), and/or some synthetic oligodeoxynucleotides (ODNs). The detection limit of AZT in the absence and in the presence of ssDNA (10 microg/ml) is 1 and 250 nM, respectively. It was found that the signal of AZT is not substantially affected by the presence of DNA. We can therefore assume that the electrons are transferred through the adsorption layer of nucleic acids. By using the elimination procedure, both irreversible reduction signals of AZT and DNA are augmented. Moreover, the elimination signal in the peak-counterpeak form may indicate the adsorption of the analytes on the electrode surface preceding an electron transfer. PMID- 15110245 TI - Application of thin film mercury electrodes and solid amalgam electrodes in electrochemical analysis of the nucleic acids components: detection of the two dimensional phase transients of adenosine. AB - The optical diffractive (DOE)-based sensor was used to the study of the optical roughness of different carbon/graphite electrodes modified by mercury film (MFEs) and solid amalgam-alloy electrodes (S-MeAEs). The electrode surfaces were visualised by an optical metallurgical microscope. The adsorption of adenosine at the MFEs and S-MeAEs has been investigated by capacitance measurement. Some kinetics aspects, such as the influence of the surface morphology, nature of the substrate and thickness of the mercury film and amalgam-alloy on the formation of two-dimensional (2D) physisorbed adenosine adlayer on the MFEs and S-MeAEs, were studied. PMID- 15110246 TI - Redox-triggered events in cytochrome c nitrite reductase. AB - Escherichia coli cytochrome c nitrite reductase is a homodimeric enzyme whose 10 heme centres range in reduction potential from ca. -30 to -320 mV. Protein film voltammetry (PFV) was performed to assess how the reactivity of the enzyme towards a number of small molecules was influenced by heme oxidation state. The experimental approach provided a high-resolution description of activity across the electrochemical potential domain by virtue of the fact that the enzyme sample was under the precise potential control of an electrode at all times. The current potential profiles displayed by nitrite reductase revealed that heme oxidation state has a profound, and often unanticipated, effect on the interactions with substrate molecules, nitrite and hydroxylamine, as well as the inhibitor, cyanide. Thus, PFV provides a powerful route to define redox-triggered events in this complex multi-centred redox enzyme. PMID- 15110247 TI - Spectroelectrochemical study of heme- and molybdopterin cofactor-containing chicken liver sulphite oxidase. AB - Electron transfer (ET) in sulphite oxidase (SOx), a heme- and molybdopterin cofactor-containing enzyme, was studied spectroelectrochemically using capillary gold electrode modified with aldrithiol. Direct electron exchange between SOx and the surface of modified gold was observed, with a formal potential of -115 mV vs. Agmid R:AgCl, KCl(sat) at pH 7.0. This value agreed well with that previously reported for redox transformation of the heme domain of SOx. However, no bioelectrocatalysis of sulphite oxidation was observed in phosphate buffer solutions. This fact evidently correlated with known inhibition of intramolecular ET in SOx by the presence of bivalent inorganic anions. After changing to a Tris buffer solution, spectra variations and cyclic voltammetry data designated direct ET-based bioelectrocatalysis of sulphite oxidation, upon addition of sulphite. Thus, the bioelectrocatalytic 2e(-) oxidation of sulphite catalysed by SOx due to direct ET exchange with the electrode was attained at aldrithiol-modified gold electrodes and shown to depend essentially on the nature of the buffer solution. PMID- 15110248 TI - Cyclic voltammetry and voltabsorptometry studies of redox proteins immobilised on nanocrystalline tin dioxide electrodes. AB - Protein film cyclic voltammetry is a well-established technique for the study of redox proteins immobilised on electrode surfaces. In this paper, we use nanostructured SnO(2) electrodes to demonstrate that cyclic voltabsorptometry is an effective, complimentary approach to such studies of protein redox function. We exemplify this approach using two different redox systems: microperoxidase-11 (MP-11) and flavodoxin Desulfovibrio vulgaris Hildenborough (Fld). Both systems were immobilised on nanocrystalline SnO(2) electrodes and the resulting films investigated by simultaneous cyclic voltammetry and voltabsorptometry. We demonstrate that cyclic voltabsorptometry allows the unambiguous and background free observation of redox reactions for both systems studied. PMID- 15110249 TI - Analysis of the interaction of a hybrid system consisting of bovine adrenodoxin reductase and flavodoxin from the cyanobacterium Anabaena PCC 7119. AB - The mitochondrial steroid-hydroxylating system in vertebrates and the NADPH producing electron transfer chain in photosynthetic organisms contain structurally and functionally similar components. Examination of a potential hybrid reconstitution of the electron transfer chain between different components of both systems could help to improve our knowledge on protein-protein interaction and subsequent electron transfer. Here we analyzed the interaction between bovine adrenodoxin reductase and flavodoxin from the cyanobacterium Anabaena PCC 7119. Optical biosensor as well as steady state and fast kinetic experiments showed their ability to form distinct productive complexes. Compared with the corresponding physiological systems the electron transfer is rather slow, probably due to the lack of specificity at the interaction surface. PMID- 15110250 TI - Study of temperature influence on electron transport in higher plants via delayed luminescence method: experiment, theory. AB - The temperature dependence of steady-state millisecond delayed luminescence (DL) is studied within the temperature range from -23 to 45 degrees C in leaf segments of chinese rose (Hibiscus rosa-sinensis L.) and bean (Ficia faba). To describe the experimental dependence of DL steady-state intensity on temperature theoretically, we suggest the temperature dependences of rate constants in earlier proposed model of photosynthesis. Under these conditions, the temperature dependence of DL steady-state value has the same form as experimental curve. PMID- 15110251 TI - Electron transfer kinetics in photosynthetic reaction centers embedded in polyvinyl alcohol films. AB - The coupling between electron transfer and protein dynamics has been studied at room temperature in isolated reaction centers (RCs) from the photosynthetic bacterium Rhodobacter sphaeroides by incorporating the protein in polyvinyl alcohol (PVA) films of different water/RC ratios. The kinetic analysis of charge recombination shows that dehydration of RC-containing PVA films causes reversible, inhomogeneous inhibition of electron transfer from the reduced primary quinone acceptor (Q(A)(-)) to the secondary quinone Q(B). A more extensive dehydration of solid PVA matrices accelerates electron transfer from Q(A)(-) to the primary photooxidized electron donor P(+). These effects indicate that incorporation of RCs into dehydrated PVA films hinders the conformational dynamics gating Q(A)(-) to Q(B) electron transfer at room temperature and slows down protein relaxation which stabilizes the primary charge-separated state P(+)Q(A)(-). A comparison with analogous effects observed in trehalose-coated RCs suggests that protein motions are less severely reduced in PVA films than in trehalose matrices at comparable water/RC ratios. PMID- 15110252 TI - Binding of single nucleotides to H+-ATP synthases observed by fluorescence resonance energy transfer. AB - F(0)F(1)-ATP synthases couple proton translocation with the synthesis of ATP from ADP and phosphate. The enzyme has three catalytic nucleotide binding sites, one on each beta-subunit; three non-catalytic binding sites are located mainly on each alpha-subunit. In order to observe substrate binding to the enzyme, the H(+) ATP synthase from Escherichia coli was labelled selectively with the fluorescence donor tetramethylrhodamine (TMR) at position T106C of the gamma-subunit. The labelled enzymes were incorporated into liposomes and catalysed proton-driven ATP synthesis. The substrate ATP-Alexa Fluor 647 was used as the fluorescence acceptor to perform intermolecular fluorescence resonance energy transfer (FRET). Single molecules are detected with a confocal set-up. When one ATP-Alexa Fluor 647 binds to the enzyme, FRET can be observed. Five stable states with different intermolecular FRET efficiencies were distinguished for enzyme-bound ATP-Alexa Fluor 647 indicating binding to different binding sites. Consecutive hydrolysis of excess ATP resulted in stepwise changes of the FRET efficiency. Thereby, gamma subunit movement during catalysis was directly monitored with respect to the binding site with bound ATP-Alexa Fluor 647. PMID- 15110253 TI - Feedback loops in the signal paths controlling gene expression. AB - Theoretical and empirical studies support the concepts suggesting emission of electromagnetic fields by living cells. Experiments performed in our laboratory indicate, in addition, tight links between attributes of these fields and the phase of cell division cycle, i.e., processes governed by gene transcription. This work supplements the now quite extensive literature concerning gene networks in that it identifies the well-known feedback mechanisms in exact topological and quantitative terms. Special diagrammatic apparatus is employed for this purpose. The simple system comprising one gene and one transcription factor (binding as a dimer to the gene regulatory region), controlled via phosphorylation process by external signal, is analyzed to demonstrate the approach. The diagram representative of the system's dynamics contains a feedback loop; the so-called transmission function of this feature of diagram topology is shown to control the dependences of gene product concentration on the stimulus intensity. PMID- 15110254 TI - Bioelectrochemical signaling in green plants induced by photosensory systems. AB - Plants generate various types of intracellular and intercellular electrical events in response to environmental stress. The generation of electrophysiological responses induced by blue and red photosensory systems was observed in soybean plants. A phototropic response is a sequence of the following four processes: reception of a directional light signal, signal transduction, transformation of the signal into a physiological response, and the production of a directional growth response. It was found that the irradiation of soybean plants at 450+/-50, 670, and 730 nm induces action potentials with duration times and amplitudes of approximately 0.3 ms and 60 mV respectively. PMID- 15110255 TI - A theoretical model for electron and proton coupling at quinone-binding site of photosystem II of higher plants. AB - In this work we consider the coupling of electron and proton transfer near Q(B) in the reaction center (RC) of photosystem II (PS2). We have carried out the calculations of the energy levels and proton density in the system Q(B)(-) Histidine L190. It is shown that the proton of the histidine forms the H-bond with twice-reduced Q(B)(2-). Based on these calculations, we propose a new explanation of the coupling between the electron and proton transfer. PMID- 15110256 TI - Photoelectrochemical study on photosynthetic pigments-sensitized nanocrystalline ZnO films. AB - Hetero-structures formed by quantum-sized ZnO nanocrystals and photosynthetic pigments were prepared by adsorbing either chlorophyll a, carotenoids or their mixture onto a film of organic-capped ZnO nanoparticles. Photoelectrochemical measurements were comparatively performed on both bulk and nanocrystalline ZnO films after dye-covering in order to probe the photosensitization process occurring at the hetero-junction. The photoconversion process was found to be greatly enhanced at the nanocrystalline electrodes upon sensitization with a dye mixture. The sensitization process is discussed on the basis of the aggregation state of chlorophyll a, and of the specific photoprotective action played by carotenoids. PMID- 15110257 TI - Enrichment of cardiolipin content throughout the purification procedure of photosystem II. AB - Photosystem II is a multisubunit membrane complex which performs the water oxidation process in the higher plants. Core dimers and monomers of photosystem II have been isolated from thylakoid membranes by sucrose density gradient centrifugation. Lipids extracted from different photosystem II-enriched fractions obtained from spinach thylakoids have been analysed by thin layer chromatography. Cardiolipin is enriched throughout the purification of photosystem II complexes; in particular dimers contained two times more cardiolipin than their monomeric counterparts. PMID- 15110258 TI - Spectroscopic and electrochemical study of Rose Bengal in aqueous solutions of cyclodextrins. AB - The interaction of Rose Bengal (RB) in aqueous solution of LiClO4 0.1 M with alpha-cyclodextrin (alpha-CD), hydroxypropyl-beta-cyclodextrins (HP-beta-CD) and hydroxypropyl-gamma-cyclodextrins (HP-gamma-CD) were studied by spectrophotometric measurements. The presence of Induced Circular Signals and the results of the analysis of the modifications in the absorbance spectra of RB produced by the presence of CDs in solution indicate that RB forms inclusion complexes only with HP-beta-CD and with HP-gamma-CD. PMID- 15110259 TI - Role of endogenous lipids in the chromophore regeneration of bacteriorhodopsin. AB - The regeneration method of Khorana [J. Biol. Chem. 262 (1987) 9271] has been modified in order to study the effect of endogenous archaeabacterial lipids and, in particular, of glycocardiolipin (GlyC) in the refolding and chromophore regeneration of bacteriorhodopsin (BR). BR refolding and chromophore regeneration could be obtained in the presence of endogenous lipid mixtures containing or not containing glycocardiolipin; however, the kinetics of bacteriorhodopsin regeneration in the presence of glycocardiolipin was faster than in its absence. These results show for the first time that the interaction of glycocardiolipin with bacteriorhodopsin favours its refolding from the denaturated state and the chromophore regeneration. PMID- 15110260 TI - Photophysical and electrochemical properties of chlorophyll a-cyclodextrins complexes. AB - In this work, we have studied the interactions between two different cyclodextrins (CDs) and chlorophyll a (Chl a) in the presence of electrolyte by means of absorption, fluorescence spectroscopy, circular dichroism and cyclic voltammetry. The results obtained indicate that the presence of both CDs gives rise to an increase of Chl a solubility in water. In particular, heptakis-(2,3,6 tri-O-methyl)-beta-cyclodextrin (TRIMEB) favours the dissolution of Chl a monomer in aqueous solution, whereas the presence of hydroxypropyl-beta-cyclodextrin (beta-HP-CD) promotes the pigment aggregation. PMID- 15110261 TI - Evaluation of chlorophyll fluorescence and membrane injury in the leaves of barley cultivars under osmotic stress. AB - Two physiological tests for screening drought tolerance of barley (Hordeum vulgare, L.) plants are compared in this work. Water deficit is induced by treating the plants' roots with polyethylene glycol (PEG 8000). The relative water content (RWC) of the plants is used as a measure of the water status. Conductometrically determined electrolyte leakage from the leaf tissue demonstrates the membrane injury caused by dehydration. It is shown that the injury index increases with the decrease of the RWC of the leaves. The F(v)/F(m) ratio is employed to assess changes in the primary photochemical reactions of the photosynthetic apparatus after dehydration. The results suggest that PSII is weakly affected by the imposed osmotic stress. The fluorescence behaviour of the examined cultivars is related to their RWC. PMID- 15110262 TI - pH-sensitive fluorescent dye as probe for proton uptake in photosynthetic reaction centers. AB - Isolated and purified reaction centers (RC) from Rhodobacter sphaeroides R-26.1 were solubilised in detergent with excess quinone and external electron donors and illuminated in the presence of pyranine. The pH change accompanying the reaction center photocycle was monitored by recording the variation of the pyranine fluorescence intensity. Using Q(B)-depleted reaction centers or blocking the photocycle with terbutryne strongly reduced the pH change. The usefulness and limits of this technique in monitoring the pH changes during the RC photocycle are also discussed. PMID- 15110263 TI - The mechanisms of lipid-protein rearrangements during viral infection. AB - Membrane fusion and fission are important events in living cell functioning. In spite of the great variety of specific cases, all of these phenomena are probably governed by the same physical principles. The first insight into physics of membrane fusion has been achieved through studies on model lipid systems. These results served as a base for subsequent investigations of the mechanisms of biological fusion. The main objective of this brief review is to expose the landmarks on the pathway of these studies and to discuss problems and perspectives. Fusion is a multistage process that includes transitions between several numbers of the intermediates. It is adopted that in the case of fusion of two planar bilayers, the following stages take place: formation of close inter membrane contact, appearance of local monolayer bridge called a stalk, expansion of stalk leading to formation of hemifusion diaphragm (HD) and, finally, creation of fusion pore. Note that the stalk is nanoscopic and still an invisible object. However, there are no doubts that some kinds of monolayer bridge exist while its shape and structure, energetic and kinetic properties are unknown. The main results on the mechanism of biological fusion were obtained on the cells expressing fusion protein of influenza virus, hemagglutinin (HA). However, this system has no M1 and M2 proteins of influenza, which are responsible for the release of the genetic material of the virus into the target cell. An experimental system developed in our laboratory allows to monitor the fusion of single virions with lipid bilayer and detect RNA release as well as the role of M1 and M2 in this process. Biological fusion is a result of complicated interplay of lipids and special proteins at nanoscopic range. It seems probable that the first function of the proteins is the preparation of a pre-fusion state also known as membrane docking. Redistribution of the energy between proteins and lipids leads to the creation of so-called dimples accumulating bending energy, which facilitates stalk formation. Probably, proteins participate in the subsequent stages of fusion in the course of a set of downhill conformational changes. Unfortunately, the data on the kinetics of these transitions are not available. Therefore, theoretical analysis is limited by a consideration of lipidic subsystem, while proteins participate as boundary conditions or some superimposed constraints. As a result, taking into account lipid tilting and fusion pore compression, low-energy pathway was proposed, leading directly from modified stalk to pore. PMID- 15110264 TI - Spectroelectrochemical studies of bilayers of phospholipids in gel and liquid state on Au(111) electrode surface. AB - Differential capacity, chronocoulometry and Polarization Modulation Fourier Transform Infrared Reflection Absorption Spectroscopy (PM FTIRRAS) were employed to investigate spreading of small unilamellar vesicles (SUVs) of DOPC and DMPC onto a Au(111) electrode surface. The electrochemical experiments demonstrated that vesicles fuse onto the electrode surface and at E>-0.5V (SSCE) or at charge densities -10-0.5 V (SSCE), the tilt angle increases to approximately 42 degrees. The increase of the tilt angle is discussed in terms of a change in the packing of the polar head of the phospholipids molecules in the bilayer adsorbed at the electrode surface. PMID- 15110265 TI - How do P-type ATPases transport ions? AB - P-type ATPases are a large family of membrane proteins that perform active ion transport across biological membranes. In these proteins, the energy-providing ATP hydrolysis is coupled to ion transport of one or two ion species across the respective membrane. The pump function of the investigated pumps is described by a so-called Post-Albers cycle. Main features of the pumping process are (1) a Ping-Pong mechanism, i.e. both transported ion species are transferred successively and in opposite direction across the membrane, (2) the transport process for each ion species consists of a sequence of reaction steps, which are ion binding, ion occlusion, conformational transition of the protein, successive deocclusion of the ions and release to the other side of the membrane. (3) Recent experimental evidence shows that the ion-binding sites are placed in the transmembrane section of the proteins and that ion movements occur preferentially during the ion binding and release processes. The main features of the mechanism include narrow access channels from both sides, one gate per access channel, and an ion-binding moiety that is adapted specifically to the ions that are transported, and differently in both principal conformations. PMID- 15110266 TI - Calcium transport by sarcoplasmic reticulum Ca-ATPase can be investigated on a solid-supported membrane. AB - Sarcoplasmic reticulum (SR) native vesicles incorporating Ca-ATPase are adsorbed on a solid-supported lipid membrane (SSM). Upon adsorption, the ion pumps are chemically activated by concentration jumps of ATP and the capacitive current transients generated by SR Ca-ATPase are measured under potentiostatic conditions. The Michaelis-Menten constant, K(M), for ATP is evaluated by varying the concentration of ATP in the activating solution. This preliminary result shows that ion transport by SR Ca-ATPase can be suitably investigated by a technique based on concentration jumps on an SSM. PMID- 15110267 TI - Tethered bilayer lipid membranes self-assembled on mercury electrodes. AB - In order to incorporate integral proteins in a functionally active state, metal supported lipid bilayers must have a hydrophilic region interposed between the bilayer and the metal. This region is realized with a hydrophilic molecule terminating at one end with a sulfhydryl or disulfide group that anchors this "hydrophilic spacer" to the surface of a metal, such as gold or mercury. The other end of the hydrophilic spacer may be covalently linked to the polar head of a phospholipid molecule, giving rise to a supramolecule called "thiolipid" (TL). With respect to gold, mercury has the advantage of providing a defect-free and fluid surface to the self-assembling spacer. Hydrophilic spacers consisting of a polyethyleneoxy or a hexapeptide chain, as well as thiolipids derived from these spacers, were employed to fabricate mercury-supported lipid bilayers. The formation of a lipid bilayer on top of a self-assembled monolayer of a hydrophilic spacer, or of a single-lipid monolayer on top of a self-assembled monolayer of a thiolipid, was realized by simply immersing the coated mercury electrode into an aqueous solution across a lipid film previously spread on its surface at its spreading pressure. Particularly stable mercury-supported lipid bilayers were obtained by using thiolipids. The biomimetic properties of these lipid bilayers were tested by incorporating channel-forming polypeptides (gramicidin and melittin) and proteins (OmpF porin). The effect of the transmembrane potential on the function of these channels was estimated by using a simple electrostatic model of the mercury-solution interphase. PMID- 15110268 TI - The study of the interaction of a model alpha-helical peptide with lipid bilayers and monolayers. AB - We studied the interaction of the alpha-helical peptide acetyl-Lys(2)-Leu(24) Lys(2)-amide (L(24)) with tethered bilayer lipid membranes (tBLM) and lipid monolayers formed at an air-water interface. The interaction of L(24) with tBLM resulted in adsorption of the peptide to the surface of the bilayer, characterized by a binding constant K(c)=2.4+/-0.6 microM(-1). The peptide L(24) an induced decrease of the elasticity modulus of the tBLM in a direction perpendicular to the membrane surface, E(radial). The decrease of E(radial) with increasing peptide concentration can be connected with a disordering effect of the peptide to the tBLM structure. The pure peptide formed a stable monolayer at the air/water interface. The pressure-area isotherms were characterized by a transition of the peptide monolayer, which probably corresponds of the partial intercalation of the alpha-helixes at higher surface pressure. Interaction of the peptide molecules with lipid monolayers resulted in an increase of the mean molecular area of phospholipids both in the gel and liquid crystalline states. With increasing peptide concentration, the temperature of the phase transition of the monolayer shifted toward lower temperatures. The analysis showed that the peptide-lipid monolayer is not an ideally miscible system and that the peptide molecules form aggregates in the monolayer. PMID- 15110269 TI - Interaction of thin wetting films of lecithin with some divalent cations. AB - The impact of some divalent cations on the structure of a model membrane system, comprising wetting lipid films, is assayed in this work. The results from impedimetry suggest prominent structural changes upon the addition of the discussed ions to the electrolyte solution contacting the film. These changes are manifested by the increase of resistivity of the films as well as by the decrease of capacitance dispersion. In accordance with other data in the literature, manganese (Mn(2+)) turned out to have an effect greater than those of Mg(2+) and Ca(2+). PMID- 15110270 TI - Shape transformation of giant phospholipid vesicles at high concentrations of C12E8. AB - Giant unilamellar phospholipid vesicles were prepared by the method of electroformation from 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphatidylcholine (POPC). We studied the influence of different concentrations of the surfactant octaethyleneglycol dodecylether (C(12)E(8)) on the spontaneous shape transformations of POPC vesicles at room temperature. In accordance with previous results, we observed that low concentration of C(12)E(8) increased the speed of the characteristic vesicle shape transformation, starting from the initial shape with thin tubular protrusion, through beaded protrusion where the number of beads gradually decreased, to final spherical shapes with invagination, whereby the average mean curvature of the vesicle membrane monotonously decreased. In contrast, higher concentration of C(12)E(8) initially induced the shape transformation in the "opposite direction": in the protrusion, the number of beads gradually increased and eventually a tube was formed whereby the average mean curvature of the vesicle membrane gradually increased. However, at a certain point, an abrupt shape change took place to yield the vesicle with invagination. In this transition, the average mean curvature of the vesicle membrane discontinuously decreased. After this transition, the vesicle began to shrink and finally disappeared. We discuss possible mechanisms involved in the observed transformations. PMID- 15110271 TI - Influence of PAMAM dendrimers on human red blood cells. AB - Polyamidoamine (PAMAM) dendrimers with different concentrations (1 nM-1 mM) (generations 2, 3, and 4) impact on human red blood cell morphology, and membrane integrity is studied. Erythrocyte shape changes from biconcave to echinocytic in dendrimers. Cell aggregation occurs. Polymers cause also concentration- and generation-dependent haemolysis. PMID- 15110272 TI - Incorporation of fluorescent probes into PAMAM dendrimers. AB - Interactions of two fluorescent probes 1-(trimethylammoniumphenyl)-6-phenyl-1,3,5 hexatriene p-toluenesulfonate (TMA-DPH) and 12-(9-anthroyloxy) stearic acid (12 AS) with polyamidoamine (PAMAM) dendrimers were studied. Changes in fluorescence intensity and steady-state fluorescence anisotropy of TMA-DPH and 12-AS were monitored. It was found that 12-AS molecules incorporated into dendrimer cavities whereas TMA-DPH molecules aggregated on the surface of polymer. Dendrimer size had not significant impact on its host properties. PMID- 15110273 TI - An integrated bienzyme glucose oxidase-fructose dehydrogenase-tetrathiafulvalene 3-mercaptopropionic acid-gold electrode for the simultaneous determination of glucose and fructose. AB - A bienzyme biosensor for the simultaneous determination of glucose and fructose was developed by coimmobilising glucose oxidase (GOD), fructose dehydrogenase (FDH), and the mediator, tetrathiafulvalene (TTF), by cross-linking with glutaraldehyde atop a 3-mercaptopropionic acid (MPA) self-assembled monolayer (SAM) on a gold disk electrode (AuE). The performance of this bienzyme electrode under batch and flow injection (FI) conditions, as well as an amperometric detection in high-performance liquid chromatography (HPLC), are reported. The order of enzyme immobilisation atop the MPA-SAM affected the biosensor amperometric response in terms of sensitivity, with the immobilisation order GOD, FDH, TTF being selected. Similar analytical characteristics to those obtained with single GOD or FDH SAM-based biosensors for glucose and fructose were achieved with the bienzyme electrode, indicating that no noticeable changes in the biosensor responses to the analytes occurred as a consequence of the coimmobilisation of both enzymes on the same MPA-AuE. The suitability of the bienzyme biosensor for the analysis of real samples under flow injection conditions was tested by determining glucose in two certified serum samples. The simultaneous determination of glucose and fructose in the same sample cannot be performed without a separation step because at the detection potential used (+0.10 V), both sugars show amperometric response. Consequently, HPLC with amperometric detection at the TTF-FDH-GOD-MPA-AuE was accomplished. Glucose and fructose were simultaneously determined in honey, cola softdrink, and commercial apple juice, and the results were compared with those obtained by using other reference methods. PMID- 15110274 TI - Chemical imaging of biological systems with the scanning electrochemical microscope. AB - A brief overview on recent advances in the application of scanning electrochemical microscopy (SECM) to the investigation of biological systems is presented. Special emphasis is given to the mapping of local enzyme activity by SECM, which is exemplified by relevant original systems. PMID- 15110275 TI - Application of integrated SECM ultra-micro-electrode and AFM force probe to biosensor surfaces. AB - The integration of scanning electrochemical ultra-micro-electrode (UME) with atomic force microscope cantilever probe have been achieved by using a homemade photolithography system. A gold-film-coated AFM cantilever was insulated with photo resist coating and a pointed end of the AFM probe was opened by illuminating with maskless arbitrary optical micro-pattern generator. To realize precise control of probe sample distance constantly, the resulting scanning electrochemical microscopy (SECM)-AFM probe was operated using a dynamic force microscopy (DFM) technique with magnetic field excitation. From a steady-state voltammetric experiment, the effective electrode diameters of the probes thus prepared were estimated to be from 0.050 to 6.2 microm. The capability of this SECM-AFM probe have been tested using gold comb in the presence of Fe(CN)(6)(3-). The simultaneous imaging of the topography and electrochemical activity of the strip electrode was successfully obtained. We also used the SECM-AFM to examine in situ topography and enzymatic activity measurement. Comparison of topography and oxidation current profiles above enzyme-modified electrode showed active parts distribution of biosensor surface. PMID- 15110276 TI - Real-time scanning tunnelling microscopy imaging of protein motion at electrode surfaces. AB - A mutant (K27C) of the blue copper protein azurin [Eur. J. Biochem. 194 (1990) 109; J. Mol. Biol. 221 (1991) 765] for orientated immobilisation on gold surfaces was analysed by scanning tunnelling microscopy (STM) both in a resting state and following the application of a short potential pulse between the tip and sample. PMID- 15110277 TI - Atomic force microscopy characterization of an electrochemical DNA-biosensor. AB - Electrode surface characteristics represent an important aspect on the construction of sensitive DNA electrochemical biosensors for rapid detection of DNA interaction and damage. Two different immobilization procedures of double stranded DNA (dsDNA) at the surface of a HOPG electrode were evaluated by MAC mode AFM performed in air. A thin dsDNA adsorbed film forming a network structure with holes exposing the electrode surface and a thick dsDNA film completely covering the electrode surface, presenting a much rougher structure, were investigated. The DNA surface characteristics and structure are discussed with respect to the degree of surface coverage. PMID- 15110278 TI - New acridone derivatives for the electrochemical DNA-hybridisation labelling. AB - In the field of DNA sensing, DNA hybridisation detection is generally performed by fluorescence microscopy. However, fluorescence instrumentation is difficult to miniaturise in order to produce fully integrated DNA chips. In this context, electrochemical detection of DNA hybridisation may avoid this limitation. Therefore, the use of DNA intercalators is particularly attractive due to their selectivity toward DNA double strand enabling DNA labelling without target chemical modification and, for most of them, to their electroactivity. We have synthesized a pyridoacridone derivative dedicated to DNA hybridisation electrochemical-sensing which presents good electrochemical reversibility, electroactivity at mild potentials and specificity toward DNA double strand. The electrochemical behaviour of this molecule has been assessed using cyclic voltammetry (CV). DNA/intercalator interactions were studied by differential pulse voltammetry (DPV) before application to hybridisation detection onto DNA sensors based on polypyrrole modified electrodes. PMID- 15110279 TI - Voltammetric behavior of DNA modified with osmium tetroxide 2,2'-bipyridine at mercury electrodes. AB - Osmium tetroxide complexes with nitrogen ligands (L) are probes of DNA structure and electroactive labels of DNA. Here adducts of single-stranded (ss) DNA with osmium tetroxide 2,2'-bipyridine (DNA-Os,bipy) were studied by cyclic voltammetry for the first time. It was found that at neutral pH DNA-Os,bipy produces three redox couples in the potential range between 0 and -1 V (peaks I-III) and a cathodic peak at about -1.3 V (peak IV). The latter peak decreased with increasing scan rate, and peaks arising from the forward and reverse scans exhibited the same direction, suggesting catalytic nature of the electrode process. We concluded that this peak corresponds to the known differential pulse voltammetric (polarographic) peak of DNA-Os,L adducts for which catalytic hydrogen evolution is responsible. In contrast, currents of cathodic peaks II and III increased almost linearly with increasing scan rate, suggesting involvement of adsorption in the electrode processes. Adsorptive stripping square-wave voltammetry was used to analyze the DNA-Os,bipy at low concentrations. It was shown that at neutral pH, peak III can offer sensitivity in the ppb range, which is only little lower than that reached by catalytic peak IV. The latter peak is, however, superior in sensitivity at acid pH values. PMID- 15110280 TI - Voltammetry of osmium-modified DNA at a mercury film electrode: application in detecting DNA hybridization. AB - Mercury film electrodes (MFE) have recently been used in nucleic acid electrochemical analysis as alternatives to the classical mercury drop ones. DNA modified with osmium tetroxide, 2,2'-bipyridine (Os,bipy) can be detected with a high sensitivity at mercury electrodes via measurements of a catalytic osmium signal. In this paper we show that mercury film on a glassy carbon electrode can be used in voltammetric analysis of Os,bipy-modified DNA. Application of the MFE as a detection electrode in double-surface electrochemical DNA hybridization assay involving osmium labeling of target DNA is demonstrated. PMID- 15110281 TI - Microanalysis of DNA by stripping transfer voltammetry. AB - A cathodic stripping transfer voltammetric procedure for trace determination of DNA and its components is described. The method is based on the DNA acid hydrolysis with subsequent electrochemical determination of released purine bases. In the first step, DNA is hydrolyzed for 30 min in 0.5 M perchloric acid at 75 degrees C. The electrochemical step involves generation of Cu(I)-purine base complex on a mercury electrode surface, transfer of electrode with accumulated complex into supporting electrolyte where voltammetric measurement is performed. Analysis is carried out in 14-microl drop volume (two-electrode connection) or in 30-microl drop (three-electrode connection) on a platinum plate, which is used as a counter electrode. Blank electrolyte contains 0.05 M borate buffer, pH 9.2 with 6.3 microM Cu(II). We could observe voltammetric signal at hydrolyzed nucleosides, nucleotides, ODN, and DNA containing purine bases. We are able to accumulate under the controlled potential and determine subnanomolar concentration of DNA corresponding to the amount of 200 pg of DNA. PMID- 15110282 TI - Electrochemical measurement of phenothiazine-interacted DNA. AB - The amount of DNA was measured by using thioridazine, which would be attached to the DNA, as an electrochemical indicator. An indicator (thioridazine) solution, a test solution (DNA solution), and a poly-l-lysine solution were successively placed on a glassy carbon electrode, and the electrode was allowed to dry; DNA was immobilized on an electrode surface by the electrostatic binding between DNA and poly-l-lysine. The electrode was immersed into a buffer solution for 15 min, and then differential pulse voltammetry (DPV) was carried out: the oxidation current peak of thioridazine was observed, and its magnitude depended on the amount of DNA in the solution which was used for preparing the electrode. It could be estimated between 0.2 microg DNA (corresponds to 630 pmol nucleotides) to 20 microg DNA (63 nmol nucleotides) from the oxidation peak current of DPV. PMID- 15110283 TI - Amperometric measurement of ds-DNA content using a peroxidase-modified electrode. AB - An enzyme electrode with a chemically amplified response for methylene blue (MB) was constructed from a glassy carbon electrode and a layer containing immobilized horseradish peroxidase (HRP). MB is reduced on the electrode but regenerated through the HRP-catalyzed reaction in the presence of H(2)O(2). The electroreduction/regeneration cycle for MB resulted in an amplified electrode response. The enzyme electrode was applied to the highly sensitive measurement of ds-DNA. The current for MB decreased in association with its complexation with DNA, and the current response caused by DNA was also amplified through the recycling processes. The detection limit of ds-DNA (from salmon testes) was as low as 5 ng ml(-1). PMID- 15110284 TI - Amperometric biosensor based on denatured DNA for the study of heavy metals complexing with DNA and their determination in biological, water and food samples. AB - Amperometric biosensor (BS) has been elaborated based on the stationary mercury film electrode (SMFE) with silver support and cellulose nitrate (CN) membrane containing immobilized single-stranded DNA (ssIDNA). The sorption isotherms and ssDNA-heavy metal binding constants have been obtained with the BS. According to these data, the chosen heavy metals form the following series of binding strength with ssIDNA: Pb(II)>Fe(III)>Cd(II). It has been found that upon the competitive adsorption, there exists practically simultaneous sorption of different ions at ssIDNA containing membrane. The method of the determination of heavy metals based on preconcentration of metal ions on the BS followed by the destruction of DNA metal complexes with ethylenediamine tetraacetate (EDTA) and voltammogram recording has been proposed. The lower limits of detectable contents are 1.0x10( 10), 1.0x10(-9) and 1.0x10(-7) mol l(-1) for Pb(II), Cd(II) and Fe(III), respectively. Heavy metals have been assayed in natural and drinking water, milk and blood serum samples even under simultaneous presence with a selectivity factor of 1:10. The effect of matrix components has been estimated. PMID- 15110285 TI - Electrochemical determination of 8-oxoguanine in the presence of uric acid. AB - The electrochemical oxidation of 8-oxoguanine (8-oxoG) in the presence of uric acid (UA) was studied by differential pulse voltammetry over a wide pH range (1 12). The results showed that both compounds follow a pH-dependent oxidation mechanism that involves two electrons and two protons corresponding to reversible charge transfer reactions. The difference between the peak potential for the oxidation of each analyte was found to be always less than 100 mV over the whole pH range, the separation being greater in the pH interval 4-7. In mixtures of both analytes, pH 6 was shown to be the best for 8-oxoG determination in the presence of uric acid, since the peak current is higher and a greater peak separation is achieved. PMID- 15110286 TI - Reversible immobilization of engineered molecules by Ni-NTA chelators. AB - Electrochemical synthesis of nickel-nitrilotriacetic acid (Ni-NTA) chelators, for subsequent immobilization of (His)(6)-tagged proteins (Photosystem II (PSII) as model molecule), on Au or Au-graphite electrodes is compared to chemical synthesis. Results show: (i) higher Ni-NTA surface density, (ii) shorter treatment time (1-12 min vs. 16 h normally needed for self-assembled monolayer (SAM)), (iii) possibility of addressing the chelator to only one Au electrode, in a sensor micro-array. PMID- 15110287 TI - Recombinant horseradish peroxidase - and cytochrome c-based two-electrode system for detection of superoxide radicals. AB - The reliable detection of a superoxide anion radical O2(*-) is complicated by its spontaneous dismutation reaction to H2O2 at acidic pHs. To simultaneously detect both O2(*-) and H2O2 produced in the course of its spontaneous dismutation, an electrochemical two-electrode system based on cytochrome c (cyt c) and recombinant horseradish peroxidase (rHRP) was applied. Therewith, a limited applicability of the cyt c system for the reliable monitoring of O2(*-) in acidic and neutral solutions was shown. It was demonstrated that both the reaction of O2(*-) dismutation to H2O2 and the reaction between the formed H2O2 and O2(*-) chemically decrease the amount of the initially present O2(*-), decreasing the sensitivity and reliability of the electrochemical detection at acidic pH. However, by appropriately varying solution pH, the concentration of O2(*-) initially injected in the system can be estimated from the analysis of calibration curves for H2O2 obtained with highly sensitive rHRP-modified electrode system at pH 6.0 and 7.0. PMID- 15110288 TI - Bi-enzyme sensor based on thick-film carbon electrode modified with electropolymerized tyramine. AB - Bi-enzyme sensor based on thick-film epoxy-carbon electrode modified with polytyramine has been developed and examined for the determination of peroxidase substrates and cholinesterase inhibitors. Polytyramine was obtained on the electrode surface by repeated scanning of the potential from +600 to +1800 mV vs. Ag/AgCl in tyramine solution. The enzymes were immobilized in the polytyramine matrix by cross-linking with glutaraldehyde. The biosensor developed provides a reliable and inexpensive way for preliminary testing of common environmental pollutants with a single sensor in accordance with assumed toxic effect by the choice of appropriate substrate and measurement conditions. The bi-enzyme sensor makes it possible to determine substituted phenols and aromatic amines in the micromolar range of their concentrations and anticholinesterase pesticides with detection limits of 0.1 (Coumaphos) and 0.03 micromol l(-1) (Chloropyrifos methyl). PMID- 15110289 TI - Properties of mixed alkanethiol-dendrimer layers and their applications in biosensing. AB - We studied the properties of mixed alkanethiol-dendrimer layers on a gold support and their application in biosensing. We showed that properties of glucose sensor can be modified using a different ratio of 1-hexadecanethiol (HDT) and poly(amidoamine) dendrimer of first generation (G1). The cyclic voltammetry in the presence of the redox couple, Fe(CN)(6)(3-)/Fe(CN)(6)(4-), was used for estimating how effectively the layer blocks the redox probe's access to the electrode surface. A scanning electrochemical microscope (SECM) was used to image the resulting distribution of the organic compounds. We found that with increasing content of dendrimers, the integrity of the layers was improved. PMID- 15110290 TI - Amperometric detection of phenolic compounds by polypyrrole-based composite carbon paste electrodes. AB - This contribution describes new composite carbon paste electrodes (CPEs) for the determination of phenolic compounds. The composite CPEs were prepared by in situ generation of polypyrrole (PPy) within a paste containing the enzyme polyphenol oxidase (PPO). The best paste composition (enzyme/pyrrole monomer/carbon particles/Nujol) was determined for a model enzyme, glucose oxidase (GOx) according to the enzymatic activity of the resulting electrodes and to the enzyme leakage from the paste during storage in phosphate buffer. The in situ electrogenerated PPy improves the enzyme immobilisation within the paste since practically no enzyme was lost in solution after 72 h of immersion. Moreover, the enzyme activity remains particularly stable under storage since the biocomposite structure conserves 80% of its activity after 1 month of storage. Following the optimisation of the paste composition, PPO-based carbon paste biosensors were prepared and presented excellent analytical properties toward catechol detection with a sensitivity of 4.7 A M(-1) cm(-2) and a response time lower than 20 s. The resulting biosensors were applied to the determination of polyphenolic compounds (e.g., epicatechin and ferulic acid). PMID- 15110291 TI - Biotinylated polypyrrole films: an easy electrochemical approach for the reagentless immobilization of bacteria on electrode surfaces. AB - Biotinylated bacteria were immobilized onto biotin/avidin modified electrode surfaces. Firstly, an electrospotting deposition method, followed by fluorescence microscopy, showed that bacteria were specifically grafted onto a gold surface. Fluorescence intensity versus the quantity of bacteria deposited on the surface was correlated, allowing determination of the microbial saturation point. Secondly, biotinylated bacteria were immobilized onto a glassy carbon macro electrode in order to assess immobilized bacterial denitrification activity. During a 7-day trial, the modified electrode completely denitrified 5 mM nitrate, with a rate of 1.66 mM/day over the first 3 days. When the same electrode was placed in fresh nitrate solution, the denitrification rate dropped to 0.80 mM/day. Crucially, the immobilized bacteria did not become detached from the electrode during the study. PMID- 15110292 TI - Nucleosides and ODN electrochemical detection onto boron doped diamond electrodes. AB - Boron doped diamond (BDD) is a promising material for electroanalytical chemistry due mainly to its chemical stability, its high electrical conductivity and to the large amplitude of its electroactive window in aqueous media. The latter feature allowed us to study the direct oxidation of the two electroactive nucleosides, guanosine and adenosine. The BDD electrode was first activated by applying high oxidizing potentials, allowing to increase anodically its working potential window through the oxidation of CH surface groups into hydroxyl and carbonyl terminations. Guanosine (1.2 V vs. Ag/AgCl) and adenosine (1.5 V vs. Ag/AgCl) could then be detected electrochemically with an acceptable signal to noise ratio. The electrochemical signature of each oxidizable base was assessed using differential pulse voltammetry (DPV), in solutions containing one or both nucleosides. These experiments pointed out the existence of adsorption phenomena of the oxidized products onto the diamond surface. Scanning electrochemical microscopy (SECM) was used to investigate these adsorption effects at the microscopic scale. The usefulness of BDD electrodes for the direct electrochemical detection of synthetic oligonucleotides is also evidenced. PMID- 15110293 TI - An electrochemical study of enzymatic oligonucleotide digestion. AB - This paper describes the synthesis and application of a novel ferrocene (Fc) label that can be efficiently attached to oligonucleotides. We demonstrate how pulse electrochemical methods can be used to measure very low concentrations of ferrocene label and, importantly, show good electroanalytical discrimination between a labelled oligonucleotide and an enzyme digested labelled oligonucleotide, in which the ferrocene label nucleotide conjugate has been released. Real time in situ analysis gives a much greater understanding of the process. Potential applications include the detection of specific nucleic acid sequences and measurement of nuclease activity. PMID- 15110294 TI - The effect of resting transmembrane voltage on cell electropermeabilization: a numerical analysis. AB - The transmembrane voltage induced due to applied electric field superimposes to the resting transmembrane voltage of the cell. On the part of the cell membrane, where the transmembrane voltage exceeds the threshold transmembrane voltage, changes in the membrane occur, leading to increase in membrane permeability known as electropermeabilization. This part of the cell membrane represents the permeabilized area through which the transport of molecules occurs. In this paper we calculated numerically the permeabilized area for different electric field strength, resting transmembrane voltage, cell shape and cell orientation with respect to the applied electric field. Results show that when the transmembrane voltage is near the threshold transmembrane voltage, the permeabilized area of the cell is increased on the anodic side and decreased on the cathodic side due to the resting transmembrane voltage. In some cases, only anodic side of the cell is permeabilized. Therefore, by using bipolar pulses, the permeabilized area can be significantly increased and consequentially also the efficiency of electropermeabilization. However, when the induced transmembrane voltage is far above the threshold, the effect of the resting transmembrane voltage is negligible. These observations are valid for different cell shapes and orientations. PMID- 15110295 TI - Electromagnetic field of extremely low frequency decreased adenylate kinase activity in retinal rod outer segment membranes. AB - Adenylate kinase activity in rod outer segment membranes of bovine retina decreased of about 55% when exposed to an extremely low frequency electromagnetic field of 75 Hz and 250 microT. The effect was independent of the time of permanence in the field. Negligible effects of the field were found on the enzymatic activity of a soluble isoform of adenylate kinase or of rod outer segment membranes solubilized with Triton, suggesting the importance of the membrane in determining the conditions of the enzyme inactivation. PMID- 15110296 TI - Excitation of vibrations in microtubules in living cells. AB - Microtubules, which are thought to be the primary organizers of the cytoskeleton, are electrical polar structures with extraordinary elastic deformability at low stress and with energy supply from hydrolysis of guanosine triphosphate (GTP) to guanosine diphosphate (GDP). At least a part of the energy supplied from hydrolysis can excite vibrations. Energy is mainly lost by viscous damping of the surrounding cytosol. Viscous damping is diminished by a slip layer which is formed by an attracted ionic charge layer and by a thin surface layer of the microtubule. Relaxation time caused by viscous damping may be several orders of magnitude greater than period of vibrations at 10 MHz. Energy supplied to the microtubule is of the order of magnitude of 10(-14) W cm(-1) (per unit length of the microtubule). PMID- 15110297 TI - Osmotically induced membrane tension facilitates the triggering of living cell electropermeabilization. AB - Very little is known about the molecular mechanisms supporting living cell membrane electropermeabilization. This concept is based on the local membrane permeability induced by cell exposure to brief and intense external electric field pulses. During the electric field application, an electro-induced membrane electric potential difference is created that is locally associated with the dielectric properties of the plasma membrane. When the new membrane electric potential difference locally reaches a critical value, a local alteration of the membrane structure is induced and leads to reversible permeabilization. In our study, we attempted to determine whether mechanical tension could modulate the triggering of membrane electropermeabilization. Change in lateral tension of Chinese Hamster Ovary cell membrane has been osmotically induced. Cell electropermeabilization was performed in the minute time range after the osmotic stress, i.e., before the regulatory volume decrease being activated by the cell. Living cell electropermeabilization was analyzed on cell population using flow cytometry. We observed that electropermeabilization triggering was significantly facilitated when the lateral membrane tension was increased. The main conclusion is that the critical value of transmembrane potential needed to trigger membrane electropermeabilization, is smaller when the membrane is under lateral mechanical constraint. This supports the hypothesis that both mechanical and electrical constraints play a key role in transient membrane destabilization. PMID- 15110298 TI - Evidence of electroconformational changes in membrane proteins: field-induced reductions in intra membrane nonlinear charge movement currents. AB - Experimental results are presented to show that a pulsed, intensive membrane potential can reduce intra membrane, nonlinear charge movement currents, which are the voltage-sensors in the voltage-dependent membrane proteins and in the excitation-contraction coupling of skeletal muscle fibers. The results indicate a possible mechanism involved in electrical injury: dysfunctions of the voltage dependent membrane proteins caused by electroconformational damages in their voltage-sensors. PMID- 15110299 TI - Comparison of the low-frequency magnetic field effects on bacteria Escherichia coli, Leclercia adecarboxylata and Staphylococcus aureus. AB - This work studies biological effects of low-frequency electromagnetic fields. We have exposed three different bacterial strains-Escherichia coli, Leclercia adecarboxylata and Staphylococcus aureus to the magnetic field (t<30 min, B(m)=10 mT, f=50 Hz) in order to compare their viability (number of colony-forming units (CFU)). We have measured the dependence of CFU on time of exposure and on the value of the magnetic field induction B(m). Viability decreases with longer exposure time and/or higher induction B(m) for all strains, but the quantity of the effect is strain-dependent. The highest decrease of the viability and the biggest magnetic field effect was observed with E. coli. The smallest magnetic field effect appears for S. aureus. From the measurement of the growth dynamics we have concluded that the decrease of the CFU starts immediately after the magnetic field was switched on. PMID- 15110300 TI - Pulse activity of populations of cortical neurons under microwave exposures of different intensity. AB - In rabbit pulse flows of populations of cortical neurons were investigated prior to, during, and after 1-min microwave irradiation (wavelength 37.5 cm, power density 0.2-40 mW/cm2). It was found that the microwave irradiation produced shifts in mean values of interspike intervals and in the number of spike bursts. Peculiarities of rearrangements of pulse flows of cortical neurons were conditioned by an intensity of exposures. PMID- 15110301 TI - Electrochemical determination of lead and glutathione in a plant cell culture. AB - In this work we established differential pulse anodic stripping voltammetry (DPASV) as the tool for analysis of lead in the plant cell culture. For the cultivation procedure, lead in Pb(II)-ethylenediaminetetraacetic acid (Pb-EDTA) chelate has been used. The detection limit of lead was found at 500 pM in phosphate buffer (pH 5.5), and 100 nM in prepared cells intracellular extract (20 pg Pb(II)/mg cells). For determination of cysteine-rich peptides, voltammetry in differential mode (DPV) in cobalt(III)-containing ammonia buffer (Brdicka reaction) was used. In this short communication, we present suitable voltammetric techniques for the physiological study of lead and thiols in plant cell culture. PMID- 15110302 TI - Effect of the leaving group on the electrodic reduction mechanism of anti Helicobacter pylori metronidazole derivatives, in aprotic and protic media. AB - Because redox properties are central to bioreductive drug activity and selectivity, six 2-methyl-5-nitroimidazole, substituted at the N1-ethyl side chain with I, Br, Cl, OAc, OMs and NH(3)(+) were synthesized and submitted to cyclic voltammetry and electrolyses, in order to define their electrodic reduction mechanism, in aprotic [dimethylsulphoxide (DMSO)+0.1 mol l(-1) tetrabuthylammonium perchlorate (TBAP)] and phosphate-buffered media, on glassy carbon electrode, in comparison with metronidazole. Three of these compounds, namely, the iodo, bromo and ammonium salt derivatives showed significant anti Helicobacter pylori (strain resistant to metronidazole) activity. All the cyclic voltammograms (CV), in aprotic medium, are similar to the one for metronidazole, except for -I, -Br and -NH(3)(+) derivatives. The CV of the N1-ethylhalide (-I, Br) 5-nitroimidazole showed more intense and irreversible first waves, even at faster sweep rates (nu<2 V s(-1)). The absence of the first wave anodic counterpart, along with analysis of the dependence of E(p), I(p) and other parameters with nu, and results from electrolysis (consumption of two electrons) showed the process to be an ECE system, with halide release, after uptake of two electrons. This behaviour represents a case of dissociative electron transfer (ET). For the ammonium salt, self-protonation mechanism was evident. The facility of reduction represented by the first wave potential and concerning the substituents is NH(3)(+)>Br>I>Cl>OMs>OH>OAc. In aqueous phosphate-buffered medium, the electrochemical behaviour of all the compounds is similar to the one of metronidazole, represented by a unique and irreversible 4e(-)/4H(+) wave. The order of reduction ease is NH(3)(+)>Br approximately OMs>I>OH>OAc. Aprotic medium allows a better discrimination between the substituents. Concerning biological activity, despite the impossibility of establishing a correlation, it has been observed that the more electrophilic compounds showed better anti-H. pylori activity. PMID- 15110303 TI - Thin film platinum cuff electrodes for neurostimulation: in vitro approach of safe neurostimulation parameters. AB - Thin film technology takes more and more importance in the development of biomedical devices dedicated to functional neurostimulation. Our research about the design of implant neurostimulating electrode is oriented toward thin film cuff electrodes based on a polyimide substrate covered by a chromium/gold/Pt film. The chromium/gold sputtered film serves as adhesion layer and current collector whereas platinum acts as an electrochemical actuator. The electrode surface has been designed to obey safe stimulation criteria (i.e. chemically inert noble metal, low electrode-electrolyte impedance, high electrochemical reversibility, high corrosion stability). The electrochemical behaviour of such platinum electrodes has been assessed and compared to a foil of platinum. Extensive in vitro characterisations of the both electrode types were carried out using AFM, SEM and electrochemical techniques. The role of enhanced surface roughness enabling high double layer capacitances to be achieved was clearly highlighted. The obtained results are discussed, with particular reference to thin film electrodes stability under in vitro electrical stimulation in NaCl 0.9% (physiological serum). Therefore, these thin film devices showed reversible PtOH formation and decomposition making them potentially attractive for the fabrication of implant stimulation cuff electrodes. PMID- 15110304 TI - Fluid flow electrophoresis model. AB - Molecular delivery via electroporation is typically done via molecular diffusion and tissue perfusion. The inherent variability in those distribution methods limits the efficacy of this medical and laboratory technique. Electrophoresis has been shown to improve the distribution and placement of the molecule [Gene Therapy 9 (2002) 1286]. This paper presents a fluid flow model for electrophoresis in tissues. Parallel plate and four-needle needle array electrodes are the electrodes modeled as the delivery devices. The parallel plate electrode produces a homogeneous distribution of the analyte but the needle array electrode creates a peak where the electric field effects diminish. PMID- 15110305 TI - Enhancement of the effectiveness of electroporation-augmented cutaneous DNA vaccination by a particulate adjuvant. AB - DNA vaccines are attracting increased attention due to multiple advantages over conventional vaccines. Attempts to improve these vaccines focus on enhancing DNA delivery and employing novel immunoadjuvants. Electroporation (EP) has emerged as an effective method for delivering DNA vaccines, significantly enhancing humoral and cellular responses. To further improve EP-augmented DNA vaccination, we used micron-size gold particles as a particulate adjuvant. DNA is not bound, or adsorbed, to the particles. Gold particles were coinjected intradermally with plasmid DNA encoding the hepatitis B virus surface antigen (HBsAg) into mice, both in the absence and presence of noninvasive EP. The particles enhanced the percentage of responding animals, and shortened the time for reaching maximal antibody titers by 2 weeks. Subtyping of the produced antibodies revealed a predominantly Th1-like response which did not change significantly with the absence or presence of particles. The particles likely function as an attractant for antigen-presenting cells (APCs), and probably do not affect EP or antigen expression to a significant extent. We conclude that micron-size gold particles injected intradermally together with DNA followed by EP give rise to an accelerated, potent immune response with a strong cellular component. This method may become important for the development of fast-acting therapeutic and prophylactic vaccines. PMID- 15110306 TI - Assessment of radiation safety instructions to patients based on measured dose rates following prostate brachytherapy. AB - PURPOSE: To validate radiation safety instructions to patients and to evaluate the potential radiation doses to members of the public after (125)I or (103)Pd prostate implantation. METHODS AND MATERIALS: Radiation dose rate measurements were made in the immediate postoperative period on 636 consecutive patients with stage T1-T2 prostate cancer who underwent transperineal (125)I or (103)Pd implantation at Memorial Sloan-Kettering Cancer Center during the period from August 1995 through January 2003. RESULTS: The mean radiation dose rate at the anterior skin surface following a prostate implant was 37 microSv/hr for (125)I and 8 microSv/hr for (103)Pd. At 30 cm from the anterior skin surface, these dose rates were reduced to 6 microSv/hr for (125)I and 3 microSv/hr for (103)Pd. At 1 m from the anterior skin surface the dose rates from both types of implants were reduced to less than 1 microSv/hr. The effect of body weight on dose rates from (125)I sources was examined for a select sub-group of patients and the measured dose rate was found to decrease with increasing body weight. In another group of patients, dose rate measurements were made on both lateral skin surfaces and were less than 16.8 microSv/hr in all cases. Assuming a 33% occupancy factor and utilizing the mean measured dose rate for (125)I, the time required to reach an effective dose equivalent limit of 5 mSv for caregivers was estimated to be 19 days on contact with the skin surface. Using a similar calculation, the lifetime doses for (125)I at a distance of 30 cm from the anterior skin surface, as well as the lifetime doses for (103)Pd on contact with the skin surface and at 30 cm from the anterior skin surface can be shown to be less than 5 mSv. CONCLUSIONS: The large number of cases available for this study permits a validation of radiation safety recommendations and provides concrete information from which the permitted exposure times following implantation can be estimated. The data support the conclusion that patients treated with these implants do not represent a radiation risk to members of the public. PMID- 15110307 TI - A comparison of the precision of seeds deposited as loose seeds versus suture embedded seeds: a randomized trial. AB - PURPOSE: Brachytherapy for prostate cancer with permanent low-dose-rate seeds has been shown to be an effective treatment for early stage prostate cancer. Due to the rapid falloff of dose, accurate seed placement is critical for optimal dosimetry. One approach to achieve optimal dosimetry is the use of seeds embedded in suture material. Seeds embedded in suture may not move after implantation as much as loose seeds. This would improve implant dosimetry. To evaluate this hypothesis a formal study was conducted in which half the gland was implanted with seeds embedded in suture and half with loose seeds. Final dosimetry is compared between both halves of the prostate. METHODS AND MATERIALS: Patients entered this Investigational Review Board approved prospective trial after completion of informed consent. At time of implant, the side of the gland to be implanted with loose as opposed to suture embedded seeds was randomly assigned. The patients then underwent intraoperative preplanned implantation. None of the preplans directed seed locations outside the prostate. Both the seeds embedded in suture and the loose seeds were implanted using needles with stylettes. At 4-6 weeks post implant, seed location was determined with CT. Both sides of the gland on CT were contoured and used for final dosimetric calculations. A cost function analysis was used to determine individual seed position deviation from intended to actual seed location. RESULTS: Eight patients were enrolled in the study. A total of 549 seeds were implanted; 240 seeds embedded in suture and 309 loose seeds. Prostate volumes ranged from 24.0-45.5 cc with a mean of 38.6 cc. The average radial deviation of the loose seeds from planned position was determined to be 3.1 mm compared with the average radial deviation of the suture embedded seeds of 3.7 mm. There was no improvement in the final dosimetry when suture embedded seeds where used. The D90 and V100 values for the half of the prostate implanted with suture embedded seeds were 71-140 (mean, 92.1) and 80.2-99.5 (mean, 89.6), respectively, for the half of the prostate implanted with loose seeds. (All D90, V100 values are % of prescription dose.) CONCLUSION: Seeds embedded in suture material do not lead to superior precision in seed deposition when compared with loose seeds, when implanted inside the prostate. PMID- 15110308 TI - Prostate brachytherapy seed migration and dosimetry: analysis of stranded sources and other potential predictive factors. AB - PURPOSE: The purpose of this study was to evaluate the influence of potential contributing factors to the incidence of seed migration and quality of prostate brachytherapy dosimetry. METHODS AND MATERIALS: Sixty patients were evaluated with day one and 3-12 month plain films of the pelvis and chest, and day 1 CT based dosimetry analysis. The incidence and types of seed migration were quantified. The seed migration outcome was evaluated with respect to source type (free vs. stranded), prostate volume, number of seeds, and needles. The day one prostate V100, V150, D90, and urethra D10 outcomes were evaluated with respect to source type, radiation type ((125)I vs. (103)Pd), prostate volume, prostate swelling, and quantity of migrating seeds. RESULTS: An increased incidence of day one and cumulative seed migration was predicted by free vs. stranded source type, with a relative risk of 6.97 and 3.08, respectively. Pulmonary and distal (toward the perineum) migration patterns were significantly reduced in the stranded group. An increased day one prostate V100, V150, D90, and urethra D10 outcome was predicted by stranded source type. CONCLUSION: Stranded source type was associated with decreased seed migration as well as higher prostate and urethra dosimetry values, resulting in an implant in which more radiation resided within the target volume, and less radiation metastasized to undesirable locations. PMID- 15110309 TI - Commentary on "a comparison of the precision of seeds deposited as loose seeds vs. suture-embedded seeds: a randomized trial" and "prostate brachytherapy seed migration and dosimetry: analysis of stranded sources and other potential predictive factors": to strand or not to strand--is this really the question? PMID- 15110310 TI - The effect of hormonal manipulation on urinary function following permanent prostate brachytherapy. AB - PURPOSE: To evaluate the effect of hormonal manipulation on catheter dependency, the resolution of urinary symptomatology, and the need for postbrachytherapy transurethral/transincisional resection (TURP/TUIP). METHODS AND MATERIALS: Seven hundred sixteen consecutive patients (median follow-up, 29 months) underwent brachytherapy for clinical T1b-T3a (1997 AJCC) prostate cancer from January 1998 through August 2002. Of the evaluated cohort, 400 patients were hormone naive, 227 received short-course cytoreductive (< or = 6 months) hormonal therapy, and 89 received extended (>6 months) hormonal therapy. An alpha-blocker was initiated prior to implantation and continued at least until the International Prostate Symptom Score (I-PSS) returned to baseline levels. Evaluated parameters included age, T-stage, preimplant I-PSS, ultrasound volume, treatment planning volume, hormonal status, supplemental external beam radiation therapy (XRT), isotope, urethral dose, total implant activity, D90, and V100/150/200. Catheter dependency and the incidence of TURP/TUIP were also evaluated. RESULTS: Six hundred fifty three patients (91.2%) had the urinary catheter permanently removed on day 0 with 15 patients (2.1%) requiring a catheter beyond 4 days. The I-PSS returned to within 1 point of the antecedent value at a median of 4 months. Sixteen patients (2.2%) underwent postimplant TURP/TUIP. A Cox regression indicated that preimplant I-PSS, supplemental XRT, planning target volume, hormonal therapy, and number of seeds were the strongest predictors for I-PSS resolution. Using all available data, the strongest predictors for I-PSS at 18 months following brachytherapy included variants of I-PSS, isotope, and days of catheter dependency. The maximum I-PSS, planning target volume, and XRT best predicted for prolonged (#10878;4 days) catheter dependency. The need for postimplant TURP/TUIP was most closely associated with days of catheter dependency and the maximum increase in I-PSS. However, when only data available prior to implantation was entered into the model, hormonal therapy predicted for postsurgical intervention. CONCLUSIONS: In this retrospective evaluation, hormonal manipulation did not statistically impact short-term or prolonged urinary catheter dependency or I-PSS at 18 months, but did influence time to I-PSS normalization and the need for postbrachytherapy surgical intervention. PMID- 15110311 TI - Efficacy of brachytherapy for prostate cancer in African Americans compared with Caucasians. AB - PURPOSE: To compare the biochemical response to prostate brachytherapy between African Americans and Caucasians in a consecutive series of patients treated at a single institution. METHODS AND MATERIALS: Between July 1995 and October 2001, 173 patients were treated with permanent (125)I seed implantation alone for presumed localized adenocarcinoma of the prostate. Twelve patients were African American and their biochemical response to treatment was compared with the 161 Caucasian patients. The patients were evaluated for biochemical recurrence according to the ASTRO consensus statement and for achieving and maintaining PSA nadirs of < or = 1.0, < or = 0.5, and < or = 0.2. Median pretreatment PSA level was 8 for the African American group and 6 for the Caucasian group. Median Gleason score for each group was 6 and no patients had palpable extraprostatic disease at the time of treatment. RESULTS: None of the African American patients have experienced biochemical recurrence compared with 7.5% of the Caucasian patients (p=0.34). The percentage of African American patients achieving and maintaining a PSA level of < or = 1.0 was 83% compared with 89% for the Caucasian patients (p=0.61). PSA nadir of < or = 0.5 was achieved in 75% of the African American patients and 81% of the Caucasian patients (p=0.52) and 50% of the African American patients experienced PSA levels of < or = 0.2 compared with 59% of the Caucasian patients (p=0.88). CONCLUSION: African American patients with prostate cancer have in general been reported to have worse prognosis compared with Caucasians. This series suggests similar outcome between African American and Caucasian patients treated with brachytherapy for prostate cancer. PMID- 15110312 TI - Technical improvement in permanent seed implantation: a two-stage brachytherapy system. Description and comparison with current technique. AB - PURPOSE: Permanent seed implantation by available techniques has modest limitations. A new, two-stage needle design and technique is described and evaluated in comparison to a conventional permanent seed technique. METHODS AND MATERIALS: The technique involves placing a stylet and sleeve initially into the all target coordinates prior to seed placement similar to temporary seed technique. The second stage involves consecutively removing the stylet from each sleeve and inserting a clear, plastic needle containing preloaded seeds into the sleeve and implanting the seeds. Fifty-six (125)I patients were treated with the two-stage technique. Comparisons were made with a cohort of 71 patients implanted using a conventional technique at the Seattle Prostate Institute. Prostate movement, surgical time, catheterization rate, and DVH postop dosimetry were analyzed. RESULTS: After an initial learning curve, the two-stage technique had surgical times similar to conventional techniques. Cephalad movement of 3-10 mm was noted in 4 (8%) patients vs. 71 (100%) patients with our conventional technique. Of the 6 (10%) patients who required Foley catheterization, 3 (5%) did so for 1 day and 3 (5%) did so for less than 3 weeks. Day 1 CT scan based dosimetry was calculated on all patients. The V100 ranged from 80-100% with a median of 92.5%. For primary cases, the V100 (<85%) was 14% for the conventional vs. 7% for the two-stage technique. No two-stage patient had a V100 <80%. The V100 values for the two-stage and conventional techniques demonstrated a possible advantage with the two-stage technique (mean V100 92.6% vs. 90.7%, [p=0.051]). The D90 for the two-stage technique ranged from 123-190 Gy with a median of 151.5 Gy for implant only and a median of 127 Gy for boost cases. The D90 values for the two-stage patients were slightly but not statistically better than the conventional technique (p=0.232). Thirty-one percent of conventional technique patients had a D90 <140 Gy vs. 22% for two-stage technique. CONCLUSION: This new two-stage brachytherapy technique may offer some advantages over conventional techniques including: simple and improved needle loading verification, less complicated and better visualization of needle placement, improved stabilization of the gland, and more consistent postoperative dosimetry. PMID- 15110313 TI - Brachytherapy in the treatment of Stage IV carcinoma of the base of tongue. AB - PURPOSE: Survival in patients with Stage IV carcinoma of the base of tongue (BOT) treated by surgery and radiotherapy remains poor. External beam radiotherapy (EBRT) and brachytherapy (BT) have been used as an alternative treatment. METHODS AND MATERIALS: Eighteen patients with Stage IV carcinoma of the BOT were treated by EBRT and BT. RESULTS: Local control is 89%. The 5-year overall (OS) and disease specific survival (DSS) rates are 52% and 67%. No neck node positive patient implanted in the neck developed cervical metastases. Two patients (11%) developed complications. CONCLUSIONS: Local regional control, survival, and complications in patients with Stage IV carcinoma of the BOT treated by EBRT and BT have been satisfactory. The use of brachytherapy for nodal metastases has eliminated the need for neck dissection. We recommend this approach in the treatment of Stage IV carcinoma of the BOT. PMID- 15110314 TI - (125)I liquid in an intracranial balloon: TG-43 formalism for an extended source. AB - PURPOSE: To present a calculation formalism for spherical homogeneous liquid brachytherapy sources and to analyze the difficulties encountered in numerical integration over non-spherical source volumes. METHODS AND MATERIALS: TG-43 formalism provides a general definition of the geometric factor for distributed source distributions. Here the geometric factor and dose for a spherical source are computed for an (125)I balloon. The errors in numerical summation of dose from point sources are assessed by computing ratios of the geometric factors of point-source and extended-source voxel volumes. RESULTS: This sphere calculation agrees well with measured dose values, showing maximum and average differences of 4.8% and 0.5% at the balloon surface. Numerical integration gives errors greater than 1% within five voxel radii of a voxel source. CONCLUSIONS: A new spherical dose calculation technique is proposed for brachytherapy treatment planning systems. Numerical integration over point source voxels is not accurate near the balloon surface. PMID- 15110315 TI - A dual-function mitochondrial transcription factor tunes out deafness. PMID- 15110316 TI - Neonatal retroviral vector-mediated hepatic gene therapy reduces bone, joint, and cartilage disease in mucopolysaccharidosis VII mice and dogs. AB - Mucopolysaccharidosis VII (MPS VII) is a lysosomal storage disease caused by deficient beta-glucuronidase (GUSB) activity. Accumulation of glycosaminoglycans (GAGs) in bone, cartilage, and synovium likely contributes to reduced mobility in untreated MPS VII individuals. We previously reported that neonatal intravenous injection of a retroviral vector (RV) expressing canine GUSB resulted in hepatocyte transduction in mice and dogs, and secreted GUSB was taken up from blood by other organs. Here we report the effect of this therapy on bone, cartilage, and joint disease. Osteocytes and bone-lining cells from RV-treated MPS VII mice had GUSB activity, resulting in a marked reduction, as compared with untreated MPS VII mice, in lysosomal storage in bone and at the bone:growth plate interface where bone elongation occurs. Although chondrocytes did not have detectable GUSB activity and had little reduction in lysosomal storage, the thickness of the growth plate was reduced toward normal. These pathological changes were likely responsible for improvements in facial morphology and long bone lengths. The synovium had reduced hyperplasia and lysosomal storage, and the thickness of the articular cartilage was reduced. Similarly, RV-treated MPS VII dogs had improved facial morphology and reduced lysosomal storage in osteocytes and synovium, but not chondrocytes. Nevertheless, the internal area of the trachea was increased, and erosions of the femoral head were reduced. We conclude that neonatal gene therapy can improve bone and joint disease in MPS VII mice and dogs. However, better delivery of GUSB to chondrocytes will be necessary to achieve more profound effects in cartilage. PMID- 15110317 TI - Non-viral delivery of the porphobilinogen deaminase cDNA into a mouse model of acute intermittent porphyria. AB - Acute intermittent porphyria (AIP), an inborn error of metabolism, results from the deficient activity of the third enzyme in the heme biosynthetic pathway, porphobilinogen deaminase (PBGD). Clinical symptoms of this autosomal dominant hepatic porphyria include episodic acute attacks of abdominal pain, neuropathy, and psychiatric disturbances. Current therapy based on intravenous heme administration is palliative and there is no way to prevent the attacks. Thus, efforts are focused on methods to replace the deficient activity in the liver to prevent the acute attacks of this hepatic porphyria. Here we explore the efficiency of a non-viral gene delivery to obtain PBGD expression in the liver of AIP transgenic mice. Four vectors were evaluated: naked DNA and DNA complexed to liposomes, polyethylenimine (PEI), and PEI-galactose, using a luciferase construct as reporter gene. The vectors were administered intravenously or directly into the portal vein with transient blood flow blockage. After tail vein injection of the DNA complexes, the liposome vector had the highest luciferase expression in lung and less in liver. When injected into the portal vein, the naked DNA had considerably higher hepatic reporter gene expression; 100 microg of naked DNA had the highest hepatic luciferase expression 24h after portal vein injection. When these vectors were used to deliver the PBGD gene into the AIP mouse model no enhancement of the endogenous PBGD activity in liver was detectable, despite the presence of the PBGD-plasmids as verified by PCR. Thus, more efficient non-viral vectors are needed to express sufficient PBGD activity over the endogenous hepatic level (approximately 30% of normal) in this murine system. PMID- 15110318 TI - Human mitochondrial transcription factor B1 as a modifier gene for hearing loss associated with the mitochondrial A1555G mutation. AB - Phenotypic expression of the deafness-associated homoplasmic A1555G mutation in the mitochondrial 12S rRNA gene varies from profound congenital hearing loss to normal hearing. It has been shown that this variability in clinical expression in most patients is due to the complex inheritance of multiple nuclear-encoded modifier genes. Human mitochondrial transcription factor B1 (TFB1M) has been proposed as a candidate for being such a modifier, since it methylates adenine residues in the adjacent loop of the A1555G mutation in the 12S rRNA gene. Polymorphic markers within and adjacent to the TFB1M gene were genotyped in 214 individuals from 41 multiplex families with the A1555G mutation of Spanish, Italian, and Arab-Israeli origin. Multipoint non-parametric linkage analysis of all families combined revealed an NPL score of 1.7 (P = 0.05), and a Lod score of 1.4 (P = 0.04). Linkage disequilibrium by the Transmission Disequilibrium Test at D6S1577, a microsatellite adjacent to TFB1M, showed preferential non-transmission of an allele to affected individuals with chi2 = 8.76; P = 0.003. Sequence analysis of the coding region of the gene and testing of all intragenic SNPs did not reveal a putative causative mutation. These data provide suggestive evidence that TFB1M is a nuclear-encoded modifier gene for phenotypic expression of the A1555G mutation, and that the effect may occur through a regulatory or splicing mutation. PMID- 15110319 TI - Identification of fatty aldehyde dehydrogenase in the breakdown of phytol to phytanic acid. AB - Phytol is a branched chain fatty alcohol, which is abundantly present in nature as part of the chlorophyll molecule. In its free form, phytol is metabolized to phytanic acid, which accumulates in patients suffering from a variety of peroxisomal disorders, including Refsum disease. The breakdown of phytol to phytanic acid takes place in three steps, in which first, the alcohol is converted to the aldehyde, second the aldehyde is converted to phytenic acid, and finally the double bond is reduced to yield phytanic acid. By culturing fibroblasts in the presence of phytol, increases in the levels of phytenic and phytanic acid were detected. Interestingly, fibroblasts derived from patients affected by Sjogren Larsson syndrome (SLS), known to be deficient in microsomal fatty aldehyde dehydrogenase (FALDH) were found to be deficient in this. In addition, fibroblast homogenates of these patients, incubated with phytol in the presence of NAD+ did not produce any phytenic acid. This indicates that FALDH is involved in the breakdown of phytol. PMID- 15110320 TI - Detection and assignment of CYP21 mutations using peptide mass signature genotyping. AB - Congenital adrenal hyperplasia (CAH) is a common inborn error of steroidogenesis. The clinical spectrum of CAH ranges from the severe classical form, which can be fatal in the newborn, to simple virilizing forms or a milder non-classical form which is often not diagnosed until puberty. Recessive mutations in the autosomal gene encoding 21-hydroxylase (CYP21) are responsible for approximately 95% of CAH cases. Since CYP21 genotype is generally predictive of the presence and severity of the disorder, accurate CYP21 genotyping is of clear medical significance. Determining the CYP21 genotype of an individual, using standard methods, is difficult due to the presence of a nearly identical pseudogene (CYP21P) in close proximity to the functional gene. To address the need for a comprehensive test for mutations in the CYP21 gene, we developed a multiplexed peptide mass signature genotyping (PMSG) assay and applied the assay to 151 DNA samples. CAH patients had been previously characterized for the 10 most common mutations. The PMSG assay detected all common mutations; in addition it identified six known rare mutations and also discovered four new mutations (two frameshifts in the first half of the gene, P42fs and S171fs, and two point mutations, H365Y and R479L). This assay has the potential to provide high-throughput, cost-effective analysis of the CYP21 gene to detect known mutations and identify novel variants in samples obtained from patients with CAH, individuals suspected to have CAH, and heterozygous carriers. PMID- 15110321 TI - New mutations in the PPBG gene lead to loss of PPCA protein which affects the level of the beta-galactosidase/neuraminidase complex and the EBP-receptor. AB - We describe the clinical findings, and the molecular and biochemical studies in an Italian family with recurrent hydrops fetalis due to galactosialidosis (GS). GS is a rare lysosomal storage disorder caused by a deficiency of the protective protein/cathepsin A (PPCA). This protein forms a high-molecular-weight complex with the hydrolases beta-galactosidase (GLB1) and neuraminidase (NEU1). By virtue of this association these two enzymes are correctly compartmentalized in lysosomes and protected against rapid proteolytic degradation. Controversial data show that PPCA is also present in a second complex, including the Elastin Binding Protein (EBP) the EBP-receptor, which is involved in elastogenesis, and NEU1. We investigated the potential role of the PPCA in both complexes. Two new genetic lesions (c60delG and IVS2+1 G > T) that lead to a frameshift and a premature stop codon were detected in the PPCA cDNA and genomic DNA of the patient. The deleterious effect of such mutations was confirmed by the complete absence of the PPCA protein on Western blots. Thus, we examined the effect of the loss of PPCA on the two protein complexes in the patient's fibroblasts. Interestingly, a reduced amount of both GLB1 and EBP proteins was detected. These data confirm that PPCA is present in two functional complexes one with GLB1 and NEU1 in the lysosomal lumen and the other with EBP at the cell surface. The reduction in GLB1 and EBP confirms that PPCA is essential for their integrity. PMID- 15110322 TI - Renal glucosuria due to SGLT2 mutations. AB - Isolated renal glucosuria results from mutations in SGLT2, which codes for an active transporter specific for d-glucose and expressed in the luminal membrane of the renal proximal tubule. In affected individuals, glucosuria leads to pursuit of hyperglycemia to exclude defects in glucose metabolism, and to investigation of renal proximal tubular function to exclude renal Fanconi syndrome. Here we present clinical and molecular data regarding a 19-year-old woman with isolated glucosuria. She was compound heterozygous for two SGLT2 mutations, i.e., a new missense mutation, T200K, and a known missense mutation, N654S. PMID- 15110323 TI - Novel mutations in CPT 1A define molecular heterogeneity of hepatic carnitine palmitoyltransferase I deficiency. AB - Liver carnitine palmitoyltransferase I (CPT I) deficiency is a rare disorder of hepatic mitochondrial long-chain fatty acid oxidation. It characteristically presents with symptoms associated with failure of ketogenesis (hypoketotic hypoglycemia). The disorder is due to mutations in the CPT 1A gene for which few patients have been characterized. We present here four novel mutations in five patients from four families with severe enzyme deficiency. Three of these are missense mutations (G465W, R316G, and F343V) and the fourth a nonsense mutation (R160X). Other than small Inuit and Hutterite populations in Canada and the Northern plains, there is complete heterogeneity of disease-causing mutations within CPT I deficient families with each demonstrating unique mutations. Because there are no easily recognizable disease-specific metabolite markers, diagnostic confirmation of this disorder requires a combination of enzymatic analysis and whole gene sequencing. PMID- 15110324 TI - The major allele of the alanine:glyoxylate aminotransferase gene: seven novel mutations causing primary hyperoxaluria type 1. AB - We describe 7 novel mutations occurring on the major allele of the human AGT gene in patients with primary hyperoxaluria type 1, an autosomal recessive disease resulting from a deficiency of the liver peroxisomal enzyme alanine:glyoxylate aminotransferase (AGT; EC 2.6.1.44). These mutations include 3 small deletions, 570delG, 744delC, and 983_988del, two splice junction mutations, IVS7-1G-->C and IVS8+1G-->T, and two nonsense mutations, R111X and W251X. We have also identified recurrences of previously identified reported mutations, 679-(IVS6+2)delAAgt, IVS8-3C-->G and 33insC. Deletion mutation 679-(IVS6+2)delAAgt has now been identified in a second Chinese patient and may be specific to that population. In contrast, 33insC has been found in patients of varying ethnic and racial backgrounds; a single vs multiple origin for this mutation is thus an intriguing question. It also appears to occur at a high frequency on the major allele. Five of the novel mutations were detected in patients who were compound heterozygotes for one of the common mis-targeting mutation, G170R or F152I, while the other two mutations occurred in the same patient. PMID- 15110325 TI - Dysmyelination in the brain of adolescents and young adults with maple syrup urine disease. AB - Maple syrup urine disease (MSUD) is associated with increased branched-chain amino acids (BCAA), their keto acids (BCKA), and acute or chronic encephalopathy. Aim of treatment is to reduce BCAA and BCKA to prevent or minimize brain dysfunction. We investigated 14 juvenile and adult patients with MSUD by means of cerebral magnetic resonance imaging (MRI) and correlated MRI changes to biochemical control measured as median plasma BCAA concentrations over 6-36 months prior to investigation. Abnormalities consisted of an increased signal in the white matter on T2-weighted images which is compatible with a disturbed water content of the white matter and dysmyelination. Areas affected most commonly were mesencephalon, brain stem, thalamus and globus pallidus; supratentorial lesions seem to be restricted to severe cases. No patient with white matter changes had acute neurological/encephalopathic symptoms indicating that the severity of dysmyelination does not correlate to acute neurotoxicity. PMID- 15110326 TI - Plasma phenylalanine concentrations are associated with hepatic iron content in a murine model for phenylketonuria. AB - Individuals with phenylketonuria (PKU) have been reported to have altered trace mineral status. In this study, we evaluated in a murine PKU model whether protein level and level of phenylalanine (PHE) restriction could modulate iron, copper, and zinc status. Fifty-four male weanling PKU and control mice were assigned to receive for 56 days an elemental low or normal protein diet; PKU mice also were assigned to receive PHE restriction (treated) or no restriction (untreated). PHE restricted mice consumed a prescribed dietary PHE to maintain plasma PHE concentrations between 120 and 480 micromol/L. PHE-unrestricted and control mice received equal amounts of dietary PHE. Intestinal and hepatic copper, iron, and zinc were measured at day 56 and fecal minerals measured at baseline and day 56. Mean plasma PHE concentrations were significantly greater in PKU PHE-unrestricted versus PKU PHE-restricted mice and control mice. Mean intestinal weights when normalized for body weight were significantly greater in PKU mice versus control mice. PKU PHE-unrestricted mice had significantly lower hepatic copper and zinc than PKU PHE-restricted mice, and significantly greater hepatic iron than control and PKU PHE-restricted mice. PKU PHE-unrestricted mice on a low protein diet had hepatic iron concentrations about 1.5 times that of the other mice. Fecal iron concentrations in all mice were significantly greater at day 56 than at baseline. No animal group effects or protein level effects were found for fecal copper, iron, or zinc contents. We conclude that hyperphenylalaninemia alters the metabolism of iron, copper, and zinc in PKU mice. PMID- 15110327 TI - Normal infant by a gestational carrier for a phenylketonuria mother: alternative therapy. AB - The consequences of pregnancies in untreated phenylketonuria (PKU) mothers are a high incidence of spontaneous abortion, intrauterine growth retardation with microcephaly, congenital malformations, and abnormal intellectual development. PKU fathers, on the other hand, produce normal children. Obviously children of PKU women and men are at least heterozygous, proving that the abnormalities produced by the PKU mothers are not genetic but "intrauterinely environmental." Exposure to the mother's metabolic abnormalities affects the fetus during the entire pregnancy. A PKU mother can produce a healthy infant if she maintains a very restricted and controlled diet before and during pregnancy. However, even the most recent reports describe a very high incidence of congenitally abnormal children of PKU mothers, hence dietary compliance is not working in all cases. A 26-year-old PKU patient with proven fertility underwent standard ovarian stimulation in preparation for oocyte retrieval. Following conventional co incubation of the oocytes and her husband's sperm, two embryos were transferred to the gestational carrier's uterine cavity, resulting in a single intrauterine pregnancy. Birth was induced at 39 weeks of gestation. The male infant weighed 3486 g. Head circumference was 36 cm and length 50.5 cm; there was no evidence of any abnormality and/or malformation. At 1 year of age, the child's growth measurements and development assessments were normal. This describes the first reported successful term pregnancy of an untreated PKU mother with the help of a gestational carrier (GC), producing a normal infant. This is an alternative method that should be offered to PKU women who are unable and/or unwilling to maintain a well controlled diet before and during pregnancy. PMID- 15110328 TI - Phosphoinositide deficiency due to inositol depletion is not a mechanism of lithium action in brain. AB - The "inositol depletion hypothesis" has been widely held to be the explanation for both the effect of lithium on brain function, apropos of its use in mood disorders, and on the impairment of development and induction of embryonic malformations in diverse organisms. The essence of the hypothesis is that a deficiency in cellular myo-inositol (Ins), secondary to lithium inhibition of inositol monophosphatase and/or multiple inositol polyphosphate phosphatase activities with trapping of Ins as inositol phosphates, leads to a depression of phosphatidylinositol (PtdIns) and a secondary impairment in inositide signaling. However, the ability of relatively low micromolar levels of Ins to reduce mammalian PtdIns synthetase activity in vivo has never been adequately tested. We have generated a lethal murine brain Ins deficiency model and measured PtdIns content using a novel MALDI-TOF MS method. Our results show that in the most severe Ins deficiency ever recorded in a mammal, the brain PtdIns levels do not decrease. We conclude that PtdIns deficiency due to "inositol depletion" is not a mechanism of lithium action in brain, and that Ins plays another unidentified role in the mammalian brain. PMID- 15110329 TI - Fabry disease: reduced activities of respiratory chain enzymes with decreased levels of energy-rich phosphates in fibroblasts. AB - Fabry disease (FD, MIM 301500) caused by a deficient activity of alpha galactosidase A is characterized by intralysosomal storage of glycosphingolipids. Main clinical features are paresthesia, hypohidrosis, angiokeratoma, renal insufficiency, and cardiovascular or cerebral complications. The exact pathogenesis is unclear. Beside mechanical storage biochemical factors might play a role. As FD is a multisystemic disorder and mitochondrial dysfunction has been described in patients with neuronal ceroidlipofuscinosis (another lysosomal storage disease) we examined mitochondrial function in fibroblasts from patients with FD. RESULTS: Activities of respiratory chain enzymes I, IV, and V were significantly (p < 0.01) lower in FD-cells. Mitochondrial recovery was unchanged as judged by the activity of the mitochondrial marker enzyme citratesynthase, cellular protein content was not significantly different. CP, ADP, and AMP concentrations were significantly (p < 0.01) lower in FD-cells. ATP was slightly, but not significantly reduced (p = 0.045). CONCLUSION: Organ dysfunction in FD may not only be explained by mechanical storage of glycosphingolipids. As in NCL, lysosomal storage material may lead to mitochondrial dysfunction with a reduction of respiratory chain enzyme activities and a subsequent drop in cellular levels of energy-rich phosphates. PMID- 15110330 TI - Dietary oligonucleotides from yeast RNA influence immune responses and resistance of hybrid striped bass (Morone chrysops x Morone saxatilis) to Streptococcus iniae infection. AB - Three feeding trials were conducted to evaluate potential immunomodulatory effects of nucleotides in the diet of hybrid striped bass. A basal diet was formulated from menhaden fish meal to contain 40% crude protein and 10% lipid. An oligonucleotide product (Ascogen P) from brewer's yeast was added to the basal formulation at the manufacturer's recommended rate of 0.5% to produce the experimental diet. Each diet was fed to four replicate groups of juvenile hybrid striped bass for seven or eight weeks in two separate trials. After Trials 1 and 2, a Streptococcus iniae bath challenge was executed to test the effects of diet on disease resistance. No significant difference in growth performance was observed between fish fed the basal and experimental diets. Body composition of whole fish, hematocrit and serum lysozyme levels were observed to be within normal ranges and not influenced by dietary nucleotides. Neutrophil oxidative radical production of fish fed the nucleotide-supplemented diet was significantly (P=0.011) higher than in fish fed the basal diet. Significantly (P<0.05) enhanced survival after exposure to S. iniae also was generally observed in fish fed the nucleotide-supplemented diet. In addition, fish fed the nucleotide-supplemented diet tended to have a higher antibody response based on microtitration agglutination; however, the difference was not statistically significant because of high variation between individual fish. Long-term (16 weeks) administration of oligonucleotides in Trial 3 failed to show enhancement of immune responses between treatments. It is concluded that dietary oligonucleotides positively influenced immune responses and resistance of juvenile hybrid striped bass to S. iniae infection. PMID- 15110331 TI - Protection of Penaeus monodon against white spot syndrome virus using a WSSV subunit vaccine. AB - Although invertebrates lack a true adaptive immune response, the potential to vaccinate Penaeus monodon shrimp against white spot syndrome virus (WSSV) using the WSSV envelope proteins VP19 and VP28 was evaluated. Both structural WSSV proteins were N-terminally fused to the maltose binding protein (MBP) and purified after expression in bacteria. Shrimp were vaccinated by intramuscular injection of the purified WSSV proteins and challenged 2 and 25 days after vaccination to assess the onset and duration of protection. As controls, purified MBP- and mock-vaccinated shrimp were included. VP19-vaccinated shrimp showed a significantly better survival (p<0.05) as compared to the MBP-vaccinated control shrimp with a relative percent survival (RPS) of 33% and 57% at 2 and 25 days after vaccination, respectively. Also, the groups vaccinated with VP28 and a mixture of VP19 and VP28 showed a significantly better survival when challenged two days after vaccination (RPS of 44% and 33%, respectively), but not after 25 days. These results show that protection can be generated in shrimp against WSSV using its structural proteins as a subunit vaccine. This suggests that the shrimp immune system is able to specifically recognize and react to proteins. This study further shows that vaccination of shrimp may be possible despite the absence of a true adaptive immune system, opening the way to new strategies to control viral diseases in shrimp and other crustaceans. PMID- 15110332 TI - Activation of the nitric oxide response in gilthead seabream after experimental infection with Photobacterium damselae subsp. piscicida. AB - Inoculation of small gilthead seabream (Sparus aurata) (30-75 g body weight) with a sublethal dose of different Photobacterium damselae subsp. piscicida (Pdp) strains (DI-21 and 94/99) induced an increase in serum concentrations of stable nitric oxide (NO) metabolites lasting from 6 h to six days post-infection, with a peak at 24 h. In contrast, no such response was detected in larger fish (150-600 g). Since the virulence of Pdp correlates with the presence of a polysaccharide capsular layer which can be induced by growing the bacteria in medium supplemented with 1% glucose (C+ forms), the effect of the presence of an enhanced capsular layer on the NO response in small fish was also evaluated. Although, all bacteria induced a similar rapid (6 h) and sustained (up to six days) NO response, serum concentrations of nitrites and citrulline were significantly increased in fish infected with the Pdp strains grown in glucose supplemented medium. When the NO response of fish infected with the C+ form of Pdp was blocked by prior injection of the inhibitor L-NAME, the LD(50) was reduced by over 10-fold and the mean time to death was also markedly reduced. Considering that (i) pasteurellosis only affects gilthead seabream with body weights below 100 g; (ii) capsulated Pdp are more resistant to the bactericidal action of NO and peroxynitrites than non-capsulated strains; and (iii) blocking the NO response of the fish results in greater susceptibility to Pdp, it seems reasonable to propose that the sustained NO response reported in this study represents a relevant protective mechanism of juvenile gilthead seabream against pasteurellosis. PMID- 15110333 TI - Changes of C-reactive protein levels in rainbow trout (Oncorhynchus mykiss) sera after exposure to anti-ectoparasitic chemicals used in aquaculture. AB - Changes of serum C-reactive protein (CRP) levels in rainbow trout (Oncorhynchus mykiss) were studied after exposure to formalin, metriphonate or potassium permanganate, which are used in aquaculture as anti-ectoparasitic chemicals. The CRP level in normal trout sera is 88+/-5 microg ml(-1) according to sandwich enzyme-linked immunosorbent assay. CRP levels increased to a maximum at six or nine days after exposure to formalin for 3.5 h at 300 ppm or 9.5 h at 30 ppm, respectively; these levels are 4.3 and 18 times higher than normal. At 18 days after treatment, the CRP level had decreased to significantly below the normal level. After exposure to metriphonate (0.4 ppm for 30 min), the CRP level increased significantly to a maximum at three days after exposure (9.9 times higher than normal), then decreased to below normal. With exposure to potassium permanganate at 40 ppm for 45 min, fish showed significantly lower CRP levels than the normal level at 14 days after exposure. Fish reared at a water temperature of 16.5-19.5 degrees C showed significantly higher CRP levels than those reared at 13 degrees C. Measurement of CRP levels in trout serum can be used as a bioindicator of the health condition of the fish. PMID- 15110334 TI - Molecular cloning and expression analysis of alpha 2-macroglobulin in the kuruma shrimp, Marsupenaeus japonicus. AB - The cDNA encoding the kuruma shrimp, Marsupenaeus japonicus alpha(2) macroglobulin (alpha(2)M) was obtained by screening a haemocyte cDNA library and 5' RACE PCR amplification. The full length cDNA of 4748 bp contains an open reading frame of 4518 nucleotides that translates into a 1505-amino acid putative peptide, with a 5'untranslated region (UTR) of 59 bp and a 3'UTR of 171 bp. The open reading frame encodes an N-terminal signal sequence of 17 residues and a mature protein of 1488 residues. The entire amino acid sequence is similar to the alpha(2)M sequences of arthropods (30-31% identity), mammals (26-27% identity) and fish (25-28% identity). The M. japonicus alpha(2)M sequence contains putative functional domains including a bait region, an internal thiol ester site, and a receptor-binding domain, which are present in mammalian alpha(2)Ms. In a healthy shrimp, the mRNA of alpha(2)M was mainly expressed in haemocytes. In addition, the expression level of alpha(2)M mRNA was dramatically increased by through time upon oral administration of peptidoglycan (PG), which is an immune stimulant. The highest expression of alpha(2)M mRNA was observed 7 days after feeding with PG. These results suggest that the shrimp alpha(2)M is an important molecule in immune system. PMID- 15110335 TI - Evaluation of biofilm of Aeromonas hydrophila for oral vaccination of Clarias batrachus--a carnivore model. AB - Biofilm of Aeromonas hydrophila was evaluated for oral vaccination of walking catfish (Clarias batrachus L.). Fish were fed with fish paste incorporating biofilm (BF) or free cells (FC) of A. hydrophila for 20 days and monitored for serum antibody production up to 60 days post-vaccination. Serum agglutinating antibody titre and relative percent survival (RPS) following challenge were found to be significantly higher in catfish fed with BF vaccine compared to that with FC. PMID- 15110336 TI - The development of the lymphoid organs of flounder, Paralichthys olivaceus, from hatching to 13 months. AB - The growth of the lymphoid organs, such as head kidney, spleen and thymus were studied in flounder, Paralichthys olivaceus Temminck & Schlegel, from hatching to 13 months of age. Except for the thymus, all organs grew as the fish grew. By 2 months of age the lymphoid organs attained their maximum relative weight. The organ weight showed a closer correlation to body weight than they did to age. The total number of leucocytes in the lymphoid organs increased with age, but the number per milligram of lymphoid organ remained constant. A micro and ultrastructural study of the lymphoid organs showed that the full development of the lymphoid organs was not achieved until the juvenile stage. The spleen and head kidney had mixed populations of "red" and "white" cells. The head kidney was more lymphoid than the spleen. The thymus involuted quickly during the first 6 months. The blood components had no obvious relationship with age or season during the period studied. PMID- 15110337 TI - Sequential study of antigen persistence and concomitant inflammatory reactions relative to side-effects and growth of Atlantic salmon (Salmo salar L.) following intraperitoneal injection with oil-adjuvanted vaccines. AB - The persistence of antigens at the injection site (area around pyloric caeca and spleen), concomitant inflammatory reaction and granuloma development were monitored at 3, 6 and 12 months following intraperitoneal injection with multivalent, oil-adjuvanted vaccines in Atlantic salmon. Parallel assessment of side-effect profiles and growth rate were also performed. Antigen persistence was examined by use of a monoclonal antibody that recognises Aeromonas salmonicida lipopolysaccharide in an immunohistochemical method for in situ identification of bacteria or bacterial fragments. The inflammatory reaction was monitored using standard histological techniques. The amount of persistent antigens and size of inflammation/adhesions were estimated semi-quantitatively. A steady decrease in the quantity of antigens at the injection site was observed from 3 to 12 months. Antigens were consistently found in inflamed tissues located in the pancreatic region. The size of inflammation increased during the first 6 months but declined thereafter. These findings suggest that persistent antigens at the injection site may act as inflammatory stimulants that induce and perpetuate the inflammatory reaction, eventually leading to adverse side-effects. PMID- 15110338 TI - Cloning, characterisation and expression of Aeromonas hydrophila major adhesin. AB - Aeromonas hydrophila, an important pathogen in fish, is believed to cause diseases by adhesive and enterotoxic mechanisms. The adhesion is a prerequisite for successful invasion. In this study, the gene of a 43 kDa major adhesin (designated as AHA1) was cloned and expressed. Nucleotide sequence analysis of AHA1 revealed an open reading frame encoding a polypeptide of 373 amino acids with a 20-amino-acid putative signal peptide (molecular weight 40,737 Da). The amino acid sequences of Aha1p showed a very high homology with the other two outer membrane proteins of A. hydrophila. Using the T-5 expression system, this major adhesin Aha1p was expressed in Escherichia coli. The purified recombinant adhesin could competitively inhibit A. hydrophila from invading fish epithelial cells in vitro. Western-blot analysis showed that this major adhesin is a very conserved antigen among various strains of Aeromonas. When used to immunise blue gourami, the recombinant adhesin could confer significant protection to fish against experimental A. hydrophila challenge. PMID- 15110339 TI - cDNA sequence encoding an 11.5-kDa antibacterial peptide of the shrimp Penaeus monodon. PMID- 15110340 TI - Molecular cloning and sequencing of shrimp (Penaeus monodon) penaeidin-5 cDNA. PMID- 15110341 TI - Oxygen reactions in high-pressure nitrogen gas studied with optical emission spectroscopy. AB - A study of some oxygen reactions in irradiated high-pressure (0.34-3.3MPa) nitrogen gas is presented. Nitrogen gas containing < or =1ppm H(2)O and O(2) was irradiated in a closed target chamber with 4.3MeV protons. Optical emission spectra of the NO-gamma (A(2)Sigma(+)-X(2)Pi), the NO-beta (B(2)Pi-X(2)Pi) and the O ((1)S-(1)D) transitions were recorded as a function of irradiation time at four different nitrogen-gas pressures. The results show a coupling between the NO gamma build-up half time and the NO-beta decay and the O ((1)S-(1)D) decay half times as a function of irradiation time. The binding of free atomic oxygen in reactions forming NO has implications for the balance of [(11)C]CO(2) versus [(11)C]CO production. PMID- 15110342 TI - Incorporation of thiosemicarbazide in Amberlite IRC-50 for separation of astatine from alpha-irradiated bismuth oxide. AB - A chelating resin was synthesized by incorporating thiosemicarbazide into Amberlite IRC-50, a weakly acidic polymer. Astatine radionuclides produced by alpha-irradiating bismuth oxide were separated using the newly synthesized chelating resin. The resin showed high selectivity for astatine. The adsorbed astatine was recovered using 0.1M EDTA at pH approximately 10. PMID- 15110343 TI - Direct measurements of radon activity in water from various natural sources using nuclear track detectors. AB - Radon activities measured in several types of natural waters from the Santos region in Brazil are presented. Makrofol E polycarbonate plastic detector was used. The detector foils were exposed to radon emanating from the water samples for 30 days in a system of two tightly coupled cups, one of which contained the detector foil and the other hosted the analyzed water sample. After irradiation and chemical etching of the plastic foils, the tracks produced by the alpha particles emitted by radon and its progeny were counted with a system consisting of an optical microscope and a video camera. The measured radon radioactivities ranged from 0.95 to 36.00 Bq/l for ground waters, from 0.30 to 0.54 Bq/l for sea waters, from 0.39 to 0.47 Bq/l for tap waters, from 0.43 to 2.40 Bq/l for river waters, and amounted to 2.35 Bq/l for water from the Santos/Sao Vicente public water supply. PMID- 15110344 TI - An X-ray Compton scatter method for density measurement at a point within an object. AB - A non-rotating method for non-intrusively determining the electron density at a point embedded within an object, without full imaging, is presented. Advantage was taken of the multienergetic nature of X-ray photons to devise a dual energy group multiple-detector scattering scheme for density determination. A measurement model that relates the density of the object to the detector response was formulated, then inverted to determine the electron density at the monitored point. Normalization factors were calculated to compensate for the wide energy distribution of the X-ray photons, and account for other system parameters. In addition, the spread in the source and detectors fields-of-view was accommodated by calculating in advance the volume of the inspection voxel. The method was experimentally applied to a variety of geometries and materials, showing that the electron density can be calculated with a reasonable uncertainty. PMID- 15110345 TI - Instrumental nuclear activation analysis (INAA) characterization of environmental air filter samples. AB - Nuclear techniques have been used in quantitations of environmental pollutants, and instrumental neutron activation analysis (INAA) has turned out to be particularly useful in the analysis of airborne suspended particles. This work describes the INAA characterization of the particulate material in the environmental samples obtained in a monitoring campaign in Mexico City's Metropolitan Area. As the types of the irradiation facilities and gamma-ray detection system impose some limitations on the possibilities of INAA analysis, the actual experimental conditions at Gamma Spectroscopy Laboratory, where the analysis was performed, had been assessed. The facilities had been found suitable for the analysis of samples from this campaign, in which 22 elements were determined. PMID- 15110346 TI - Radiosynthesis of 4-[(2-chloroethyl)(2-[(11)C]ethyl)amino]-phenoxycarbonyl-l glutamic acid a half mustard prodrug as a potential probe for imaging antibody- and gene-directed enzyme prodrug therapy with positron emission tomography. AB - The potential antibody directed prodrug therapy half-mustard prodrug 4-[(2 chloroethyl)(2-ethyl)amino]-phenoxycarbonyl-L-glutamic acid was synthesised by reductive alkylation of 4-[(2-chloroethyl)amino]-phenoxycarbonyl-L-glutamic acid using acetaldehyde. 4-[(2-chloroethyl)[(11)C](2-ethyl)amino]phenoxycarbonyl-L glutamic acid was synthesized with 18-22% decay corrected radiochemical yield in 45 min from EOB by reductive alkylation of 4-[(2-chloroethyl)amino] phenoxycarbonyl-L-glutamic acid using [(11)C]acetaldehyde. [(11)C]Acetaldehyde was prepared in 60% decay corrected radiochemical yield by oxidation of [(11)C]ethanol over heated copper oxide. The radiosynthesis of [(11)C]ethanol was re-examined and optimized. 4-[(2-chloroethyl)(2-ethyl)amino]-phenoxycarbonyl-L glutamic acid was found to have affinity for carboxypeptidase G2; the K(m) and V(max) were 99.4-115.9 microM (n=3) and 3.6-5.0 microM/min, respectively, at a carboxypeptidase G2 concentration of 0.0247 U/ml. PMID- 15110347 TI - Highly efficient automated synthesis of [(11)C]choline for multi dose utilization. AB - [(11)C]Choline has been under investigation as a PET ligand for imaging tumor tissue, especially prostate cancer. An improved, automated synthesis of the tracer now was established. [(11)C] Choline was produced by labeling 2 (dimethylamino)-ethanol (DMAE) with [(11)C]CH(3)I in a Tefzel tube at room temperature without solvent. The product was purified using a cation exchange cartridge. Reaction conditions were optimized with respect to synthesis time and amount of DMAE, resulting in radiochemical yields higher than 80% using 60 microl of DMAE in 20 min, radiochemical purity was >99% and residual DMAE was below 10 ppm. After (11)C-production of 1h at 50 microA [(11)C]choline activities of 30.0+/-5.6 GBq (n = 29) were obtained in sterile solution ready for intravenous administration. PMID- 15110348 TI - 2-[(18)F]-fluoroisonicotinic acid hydrazide: biological evaluation in an acute infection model. AB - We have synthesized 2-[(18)F]-fluoroisonicotinic acid hydrazide by nucleophilic displacement reaction on ethyl-2- (trimethylammonium)-isonicotinate precursor in acetonitrile. Kryptofix 222 was used as the phase transfer catalyst. The intermediate fluorinated ethyl ester reacted with hydrazine hydrate to produce the hydrazide. Excellent radiochemical yield was attained with total synthesis time of approximately 60 min. Biological evaluation was performed in bacterial cells and biodistribution in normal as well as E. coli infected CBA/J mice. It was found that the S. pneumoniae cells retained the radiotracer in an in vitro assay. The tracer showed positive localization at the infection/inflammation site in E. coli infected mice. PMID- 15110349 TI - Synthesis and biodistribution of (99m)Tc-carbonyltechnetium-labeled fatty acids. AB - In order to image myocardial metabolism, (99m)Tc(CO)(3)(+) complexes containing fatty acids were evaluated. At first, 11-(1-imidazolyl)-undecanoic acid (IUA) and 11-(2-(4-bromo-phenylazo)-1-imidazolyl)-undecanoic acid (BPIUA) were synthesized and the corresponding carbonyltechnetium-labeled complexes were prepared. However, unfortunately similar to those of other technetium-labeled fatty acid analogs, the biodistribution studies of (99m)Tc(CO)(3)-IUA and (99m)Tc(CO)(3) BPIUA in mice demonstrated poor heart-to-blood ratios, thus these complexes cannot be used as heart-imaging reagents. In addition, Tween-80 can improve the biodistribution of the complexes for myocardial imaging. PMID- 15110350 TI - Potent antiviral agent WIN 54954: high specific activity labelling with tritium. AB - Valuable antiviral compound WIN 54954 was tritiated at high specific activity and the product was characterized. PMID- 15110351 TI - Simulation of a radiotracer experiment by flow and detection-chain modelling: a first step towards better interpretation. AB - The injection of a gamma-emitting tracer in backward-facing step flow was modelled. Flow and concentration fields were calculated by CFD and detector responses by a Monte-Carlo code. The objective was to show the influence of the radiotracer energy on the detector response, so as to interpret better data from radiotracer experiments. The radiotracer selection is important in obtaining reliable results, each specific case requiring careful analysis of the experimental data. However, more work is still required to achieve the prescribed goal aimed at. PMID- 15110352 TI - Intercomparison experiments of systems for the measurement of xenon radionuclides in the atmosphere. AB - Radioactive xenon monitoring is one of the main technologies used for the detection of underground nuclear explosions. Precise and reliable measurements of (131m)Xe, (133g)Xe, (133m)Xe, and (135g)Xe are required as part of the International Monitoring System for compliance with the Comprehensive Nuclear Test-Ban Treaty (CTBT). For the first time, simultaneous testing of four highly sensitive and automated fieldable radioxenon measurement systems has been performed and compared to established laboratory techniques. In addition to an intercomparison of radioxenon monitoring equipment of different design, this paper also presents a set of more than 2000 measurements of activity concentrations of radioactive xenon made in the city of Freiburg, Germany in 2000. The intercomparison experiment showed, that the results from the newly developed systems agree with each other and the equipment fulfills the fundamental requirements for their use in the verification regime of the CTBT. For 24-h measurements, concentrations as low as 0.1 mBqm(-3) were measured for atmospheric samples ranging in size from 10 to 80 m(3). The (133)Xe activity concentrations detected in the ambient air ranged from below 1 mBqm(-3) to above 100 mBqm(-3). PMID- 15110353 TI - Experimental investigation of mass efficiency curve for alpha radioactivity counting using a gas-proportional detector. AB - Gross alpha counting of evaporated water residues offers a simple method for screening alpha radioactivity in water for both public health and emergency purposes. The evaporation process for water has been improved by using a combination of roughening of the surface of counting planchettes, two-stage evaporation, and temperature-controlled block heating. The efficiency of the gas proportional detector for alpha-particle detection in water residues was studied as a function of sample mass-thickness in the range between 0.1 and 13 mg cm(-2). The effect of alpha energy on the efficiency, as well as moisture absorption on the samples, were studied using (230)Th, (238)U, (239)Pu, (241)Am, and (244)Cm radionuclides. Also, alpha-to-beta crosstalk was investigated as a function of sample mass for (230)Th, (238)U, (239)Pu, (241)Am, and (244)Cm. The improved method can also be applied for gross alpha detection in biological fluids. PMID- 15110354 TI - Aqueous liquid scintillation counting with fluor-containing nanosuspensions. AB - A microemulsion comprised of water, Brij 78, pentanol and styrene into which PPO and bis-MSB had been dissolved was prepared. Polymerization of the styrene resulted in a suspension of fluor-containing polystyrene nanoparticles (<100 nm). After a concentration step, the aqueous nanosuspension was able to detect (14)C with counting efficiencies over 50% of those of a commercially available scintillation cocktail. Monte Carlo calculations demonstrated that the size and concentration of the nanoparticles were appropriate for optimum detection efficiency. PMID- 15110355 TI - Implementation of CIEMAT/NIST LSC efficiency tracing method in KRISS: (204)T1 standardization. AB - The CIEMAT/NIST efficiency tracing method has been widely used for radionuclide standardization. The method consists of the theoretical calculations of counting efficiencies and the experimental data obtained from a commercially available double photomultiplier detector. We have recently implemented the technique using (3)H as a tracer. Our implementation of the method was validated by participating into an international comparison of activity measurements of a solution of (204)T1. We report in this paper the procedure employed to implement the technique and compare our measurement result of (204)T1 activity with that of the international comparison. PMID- 15110356 TI - Experimental studies and nuclear model calculations on proton-induced reactions on (nat)Se, (76)Se and (77)Se with particular reference to the production of the medically interesting radionuclides (76)Br and (77)Br. AB - Excitation functions of the reactions (nat)Se(p,x)(75,76,77,82)Br, (76)Se(p,xn)(75,76)Br, (76)Se(p,x)(75)Se and (77)Se(p,xn)(76,77)Br were measured from their respective thresholds up to 40 MeV, with particular emphasis on data for the production of the medically important radionuclides (76)Br and (77)Br. The conventional stacked-foil technique was used. The samples were prepared by a sedimentation process. Irradiations were performed using the compact cyclotron CV 28 and the injector of COSY, both at the Research Centre Julich. In order to validate the data, nuclear model calculations were performed using the code ALICE IPPE which is based on the preequilibrium-evaporation model. Good agreement was found between the experimental and theoretical data, except in the high-energy region where the calculated data were somewhat higher. All the measured excitation curves were compared with the data available in the literature. From the experimental data the theoretical yields of all the investigated radionuclides were calculated and plotted as a function of proton energy. The calculated yield of (77)Br from the (nat)Se(p,x)(77)Br process over the energy range E(p)=25-->15 is 72.7 MBq/microAh and from the (77)Se(p,n)(77)Br reaction over E(p)=15-->6 MeV it is 86.2 MBq/microAh. The yield of (76)Br from the (76)Se(p,n)(76)Br reaction for E(p)=15-->8 is 360.1 MBq/microAh and from the (77)Se(p,2n)(76)Br reaction for E(p)=28-->18 MeV it is 879.2 MBq/microAh. The radionuclidic impurity levels are discussed. PMID- 15110357 TI - Experimental studies on the proton-induced activation reactions of molybdenum in the energy range 22-67 MeV. AB - The production cross-sections of (99,93m)Mo, (96,95,95m,94)Tc, (96,95,92m,90)Nb, (89,88,86)Zr and (88,87,86)Y radionuclides for proton-induced reactions on molybdenum were measured with molybdenum targets of natural isotopic composition using a stacked-foil activation technique in the energy range 22-67 MeV. The thick target integral yields were also deduced for each reaction using the measured cross-sections from the respective threshold up to 67 MeV. The results have given new data for all of the investigated radionuclides. The results of the present experiment showed excellent agreement with the earlier reported data in the lower energy region. PMID- 15110358 TI - An 82Rb infusion system for quantitative perfusion imaging with 3D PET. AB - A (82)Rb infusion system is described with two important features for imaging with 3D positron emission tomography. First, a generator bypass line is added to flush the patient infusion line at the end of an elution. Second, feedback control is implemented to permit 'slow-bolus' constant-activity elutions. A model for the activity eluted from a (82)Sr/(82)Rb generator based on a volume-activity empirical relationship, is used as the basis for performing simulations to demonstrate the efficacy of varying the flow rate through the generator to achieve desired eluted (82)Rb activity rate profiles. A (82)Rb infusion system was constructed to verify the accuracy of the simulations. The system can deliver accurate constant-activity elutions from 10% to 70% of the total generator activity. PMID- 15110359 TI - Estimation of X-ray beam quality by electron paramagnetic resonance (EPR) spectroscopy. AB - A novel dosimetry-based technique using EPR spectroscopy to determine X-ray beam quality is proposed. The radiation-sensitive material is made of a mixture of two polycrystalline substances with different X-ray absorption properties. The composite samples, consisting of polycrystalline lithium formate monohydrate and calcium formate, were prepared as pellets, X-irradiated, and analyzed with EPR spectroscopy. The ratio of the EPR signal amplitudes of the two constituents can serve as a measure of the X-ray beam quality given by the equivalent photon energy. The calculation of the signal amplitude ratio involves a reconstruction of the composite EPR spectrum. The logarithm of the signal amplitude ratio appears to be linearly correlated with the logarithm of the equivalent photon energy. The linear relationship can be used as a calibration for estimating the equivalent photon energy from the composite EPR spectrum. The composite material was used to investigate the changes in the equivalent photon energy in a Perspex phantom with increasing depth. When a 220 kV X-ray beam with an equivalent photon energy of about 100 keV was used, changes in the EPR signal amplitude ratio revealed a buildup of scattered photons with increasing depth in the phantom. This change could be related to the equivalent photon energy using the logarithmic calibration curve. It was found that the equivalent photon energy at the depth of 13 cm in the phantom was 25% lower than on its surface. The proposed method can be used for estimating equivalent photon energy in both standardized and non-standardized situations, the latter corresponding to beam setups where use of filters and ionization chambers is difficult or impossible. Also, the system can provide a means for measuring photon energy in X-irradiated phantoms. PMID- 15110360 TI - Formation of short-lived positron emitters in reactions of protons of energies up to 200 MeV with the target elements carbon, nitrogen and oxygen. AB - Excitation functions were measured by the stacked-foil technique for proton induced reactions on carbon, nitrogen and oxygen leading to the formation of the short-lived positron emitters (11)C (T(1/2) = 20.38 min) and (13)N (T(1/2) = 9.96 min). The energy region covered extended up to 200 MeV. The product activity was measured non-destructively via gamma-ray spectrometry. A careful decay curve analysis of the positron annihilation radiation was invariably performed. The experimental results were compared with theoretical data obtained using the modified hybrid nuclear model code ALICE-IPPE for intermediate energies. The agreement was found to be generally satisfactory. The data are of importance in proton therapy. PMID- 15110361 TI - Simulation of alpha particle spectra from aerosol samples. AB - A Monte Carlo code is developed for simulating energy spectra of alpha particles from aerosol samples. Geometrical detection efficiency and energy loss of the alpha particles in the source itself and in the material between the source and detector are simulated. Different characteristics of the aerosol particles and medium material are taken into account in the computation. An excellent agreement with earlier results and measurements is found. The code can be applied for example in designing optimal aerosol sampling methods in direct alpha spectrometry. PMID- 15110362 TI - Measurement of depth distributions of (3)H and (14)C induced in concrete shielding of an electron accelerator facility. AB - The estimation of radioactivity induced in concrete shielding is important for the decommissioning of accelerator facilities. Concentrations of (3)H and (14)C in the concrete shielding of an electron linear accelerator were measured, and the depth distributions of (3)H and (14)C and gamma-ray emitters were discussed in relation to their formation reactions. PMID- 15110363 TI - Determination of 4.438 MeV gamma-ray to neutron emission ratio from a (241)Am (9)Be neutron source. AB - Gamma-ray (gamma-ray) spectra of a 1.49 x 10(11)Bq (241)Am-(9)Be source and background were measured using a 2 in x 2 in, NaI(Tl) detector. Backgrounds due to the neutron interactions and energy deposition were calculated with MCNP4C. By subtracting the backgrounds from the experimental spectra, the S(gamma) is obtained and the R=S(gamma)/S(n) was estimated. The final result of R=S(gamma)/S(n)=0.596 is in agreement with result reported in literature. PMID- 15110364 TI - Diffusion of Eu(III) in compacted bentonite-effect of pH, solution concentration and humic acid. AB - The effect of pH, Eu(III) solution concentration and humic acid on the diffusion of Eu(III) in compacted bentonite (rho(b) = 1000 +/- 30 kg/m(3)) was studied with "in-diffusion" method at an ionic strength of 0.1M NaClO(4). The results (K(d) values from the first slice and theoretical calculation, apparent and effective diffusion coefficients) derived from the new capillary method are in good agreement with the literature data under similar conditions, and fit the Fick's second law very well. The results suggest that the diffusion of Eu(III) is dependent on pH values and independent on solution concentration in our experimental conditions. Humic acid forms precipitation/complexation with Eu(III) at the surface of compacted bentonite and thus deduces the diffusion/transport of Eu(III) in compacted bentonite. The K(d) values in compacted bentonite are in most cases lower than those in powdered bentonite obtained from batch experiments. The difference between the K(d) values from powdered and compacted bentonite is a strong function of the bulk density of the bentonite. The results suggest that the content of interlaminary space plays a very important role to the diffusion, sorption and migration of Eu(III) in compacted bentonite. PMID- 15110365 TI - Quantification of the dependency of radon emanation power on soil temperature. AB - A study on the quantification of the dependency of the radon emanation power on soil temperature has been made using the accumulation method. The emanation from dry soil was studied at temperatures between -20 degrees C and 45 degrees C. A formula to calculate the radon emanation power as a function of the temperatures has been developed. The formula would contribute to the modeling of radon transport in soils and building materials. PMID- 15110366 TI - Sometimes a bit of regulation can be a good thing... PMID- 15110367 TI - The effect of cisatracurium and rocuronium on cisatracurium precurarization and the priming principle. AB - STUDY OBJECTIVE: To demonstrate the effect of administering a precurarizing dose of cisatracurium or rocuronium on the speed of onset of cisatracurium, and to review the possible mechanisms and value of the priming principle. DESIGN: Double blind, randomized, controlled trial. SETTING: Inpatient anesthesia in a university teaching hospital. PATIENTS: 90 ASA physical status I and II patients undergoing elective surgery requiring endotracheal intubation. INTERVENTIONS: Three groups of 30 patients each were investigated. Following induction of anesthesia with fentanyl and propofol, Group 1 received cisatracurium 0.015 mg.k( 1), Group 2 received rocuronium 0.09 mg. kg(-1), and Group 3 (control) received normal saline. Six minutes after priming, Groups 1 and 2 received cisatracurium 0.135 mg. kg(-1) whereas Group 3 received cisatracurium 0.15 mg. kg(-1). MEASUREMENTS AND MAIN RESULTS: In each group, first twitch height and the train of-four ratios were recorded every 10 seconds after the initial priming dose. Intubation was attempted after the first twitch height became less than 15% of baseline. The decrease in the train-of-four ratios at 6 minutes was 0.97 for cisatracurium and 0.85 for rocuronium. The onset of muscle relaxation was significantly faster after priming with cisatracurium and rocuronium (71.7 +/- 21.3 and 65 +/- 19.8 sec, respectively) compared with control (148.7 +/- 43.1 sec). Females receiving both muscle relaxants had a faster onset of paralysis than did males (65.9 +/- 20.6 vs. 79.2 +/- 20.6 and 55 +/- 14.5 vs. 71.7 +/- 20.4 sec). Intubation conditions were either excellent or satisfactory in all patients. CONCLUSIONS: Six minutes after precurarization, there is no significant difference between rocuronium and cisatracurium when used as priming drugs. An even faster onset time with both drugs was demonstrated in females. The use of priming doses of 25% to 30% of ED(95) may cause symptomatic muscle weakness. The mechanisms of the priming principle are discussed. PMID- 15110368 TI - The effectiveness of continuous epidural infusion of low-dose fentanyl and mepivacaine in perioperative analgesia and hemodynamic control in mastectomy patients. AB - STUDY OBJECTIVES: To investigate, in mastectomy patients, the effectiveness of continuous cervical epidural block using a low-dose fentanyl infusion in combination with local anesthetics. DESIGN: Prospective, observational study. SETTING: 450-bed, university-affiliated hospital. PATIENTS: 21 ASA physical status I and II female patients undergoing modified radical mastectomy. INTERVENTIONS: An epidural catheter was inserted at the C(7)-Th(1) interspace before the induction of anesthesia. Anesthesia was maintained using a low concentration of sevoflurane with nitrous oxide-oxygen (N(2)O-O(2)). A mixture of 100 microg fentanyl and 49 mL of 1% mepivacaine was prepared, and 7 mL of this solution was epidurally injected before the initial incision. This same solution was continuously infused at a rate of 7 mL/hr (fentanyl 17.5 microg/hr) throughout the anesthesia, and at 2 mL/hr (fentanyl 5 microg/hr) postoperatively. MEASUREMENTS AND MAIN RESULTS: Intraoperative mean arterial pressure (MAP) and heart rate (HR), postoperative pain and analgesic use, and the frequency of postoperative side effects of anesthesia, including nausea, dizziness, and respiratory depression, were recorded. The protocol described provided stable intraoperative hemodynamic control with no or low-dose nicardipine infusion. Sufficient postoperative analgesia was achieved in 18 of 21 patients. One patient reported postoperative nausea, and no other side effects were reported. CONCLUSIONS: Continuous epidural infusion of the low-dose fentanyl mixture described above provides adequate intraoperative hemodynamic control and postoperative pain relief, with a low rate of side effects in mastectomy patients. PMID- 15110369 TI - Bacterial contamination of epidural catheters: microbiological examination of 502 epidural catheters used for postoperative analgesia. AB - STUDY OBJECTIVE: To investigate the frequency of bacterial colonization of epidural catheters used for postoperative pain treatment longer than 24 hours in abdominal, thoracic, or trauma surgery patients. DESIGN: Retrospective study. SETTING: Intermediate care facility and general ward of a university hospital. PATIENTS: 502 patients who received epidural catheters after abdominal, thoracic, or vascular surgery at our institution from January 1996 to December 2000. INTERVENTIONS: Placement of an epidural catheter, which was used for postoperative pain treatment, for more than 24 hours. The puncture site dressing included saturation each day with povidone-iodine. MEASUREMENTS AND MAIN RESULTS: Microbiologic monitoring of epidural catheter tips and daily examination of puncture sites with regard to signs of inflammation took place. Four times daily patients were examined to check adequacy of pain treatment and neurologic deficits. Catheter tip cultures were positive in 29 patients (5.8%). Staphylococcus epidermidis was isolated in 22 cases (76%). No case of spinal epidural abscess was observed within 6 months after epidural catheterization. The average catheterization time was 5 days (quartile range: 4 to 6 days). CONCLUSIONS: Meticulous management ensures a relatively low level of bacterial contamination in epidural catheters applied for postoperative pain treatment greater than 5 days. Contamination rarely leads to spinal epidural infection. PMID- 15110370 TI - Effect of single-dose fentanyl on the cardiorespiratory system in elderly patients undergoing cataract surgery. AB - STUDY OBJECTIVES: To evaluate the effects of fentanyl on the cardiorespiratory system in elderly patients undergoing cataract surgery with phacoemulsification method. DESIGN: Randomized, prospective, double-blind study. SETTING: University hospital. PATIENTS: 70 ASA physical status I, II, and III patients (>60 years) who underwent cataract surgery with topical anesthesia. INTERVENTIONS: Patients were randomly divided into two groups. The fentanyl group (35 patients) received fentanyl in 0.7 microg/kg bolus doses in a 2-mL balanced salt solution prior to surgery. The control group (35 patients) received a 2-mL balanced salt solution without any analgesic drug. MEASUREMENTS AND MAIN RESULTS: Systolic (SBP), diastolic (DBP), mean arterial pressure (MAP), heart rate (HR), peripheral oxygen saturation (SpO(2)), respiratory rate (RR), end-tidal carbon dioxide (ETCO(2)), inspired CO(2) concentration, and sedation scores were measured preoperatively and at 5, 10, 15, 20, and 30 minutes intraoperatively. Postoperatively, patients were questioned about the presence of intraoperative pain. In the fentanyl group, no significant differences were observed in SPB, DBP, MAP, RR, or peripheral SpO(2). In the control group, RR was higher than baseline values at 10, 15, and 20 minutes. Diastolic blood pressure was higher than baseline values at 20 minutes. End-tidal CO(2) and inspired CO(2) levels were higher than baseline levels in both groups at all measurement times. Intraoperative ETCO(2) levels were higher in the fentanyl group than the control group (p < 0.01). Finally, no hypoxemia was observed in either group. CONCLUSION: Fentanyl can be used safely in 0.7-microg/kg dosages in elderly patients to improve patient comfort without any cardiorespiratory side effects, when undergoing cataract surgery with topical anesthesia. PMID- 15110371 TI - Determination of right ventricular function by transesophageal echocardiography: impact of proximal right coronary artery stenosis. AB - STUDY OBJECTIVE: To investigate whether transesophageal echocardiography (TEE) can provide accurate information on right ventricular (RV) function in patients with right coronary artery (RCA) stenosis, given that a decrease in blood supply from the RCA may invalidate the use of single 2-D echocardiography imaging plane as a guide to RV function. DESIGN: Prospective, nonblinded study. SETTING: University hospital. PATIENTS: 30 adult patients undergoing elective cardiac or vascular procedures. INTERVENTIONS: Patients were classified into two groups according to the presence or absence of the proximal RCA (segment 1 or 2) stenosis. Group A patients had no obstructive lesions in the proximal RCA (n = 15). Group B patients had 75% or greater obstructive lesions in the proximal RCA (n = 15). MEASUREMENTS AND MAIN RESULTS: After induction of anesthesia, RV function was evaluated by both fast-response thermodilution pulmonary artery catheter and TEE. Transesophageal echocardiography-derived RV fractional area change (FAC) and tricuspid annular plane systolic excursion ratio (TAPSE ratio) were compared with thermodilution-derived RV ejection fraction (EF) using linear regression analysis. Transesophageal echocardiography-derived RV end-diastolic area (EDA) was compared with thermodilution-derived end-diastolic volume (EDV). Both methods showed a good correlation in RV, EDV, and EF in Group A, but no correlations in Group B. CONCLUSIONS: Transesophageal echocardiography does not provide reliable information on RVEF and EDV when proximal RCA stenosis is present. PMID- 15110372 TI - Pressure on the face while in the prone position: ProneView versus Prone Positioner. AB - STUDY OBJECTIVE: To measure the surface pressure on the face of a patient placed in the prone position with the most commonly used prone positioning devices, a non-face-contoured positioner (PP) and a new face-contoured device (PV). DESIGN: Prospective, randomized comparison. SETTING: Operating room in an American academic medical center. SUBJECTS: 35 randomly recruited adult volunteers. INTERVENTIONS: Surface pressure on the face was measured in awake subjects placed in the prone position, with the head and neck in the position of most comfort, using both the PP and PV devices. MEASUREMENTS: Surface pressure was obtained using an array of small transducers imbedded in a thin cushion that was interfaced between the face and positioning device. The amount of extension or flexion of the head on the neck was estimated using an angular measurement of eye ear line and horizontal line. MAIN RESULTS: The average surface pressure on the face was less with the PV than with the PP (21 +/- 3 mmHg vs. 27 +/- 5 mmHg; p < 0.0001). The number of areas where pressure exceeded 30 mmHg and 50 mmHg was lower for the PV than the PP (15 +/- 7.5 areas vs. 19 +/- 7.2 areas > 30 mmHg; p < 0.05; 5.2 +/- 3.3 areas vs. 9.0 +/- 5.0 areas > 50 mmHg; p < 0.0001). Pressure on the chin increased with extension of the head or neck (p < 0.05) with both devices. CONCLUSIONS: Surface pressure on the face in the prone position is 29% higher with the non-face-contoured PP than with the face-contoured PV. The number of areas on the face where the surface pressure is greater than 50 mmHg is 80% higher with the PP than the PV. Small degrees of head extension increases pressure on the chin. Both devices produce areas of pressure, typically over the chin, which may be associated with local skin damage. Keeping the head and neck in a non-flexed, non-extended position may minimize pressures. PMID- 15110373 TI - Peripheral nerve stimulators can inhibit monitor display of pacemaker pulses. AB - The operating room monitors that are currently available use digital signal processing techniques to acquire and display electrocardiograms (ECGs). As a result, pacemaker impulses are interpreted as noise and thus filtered from the final display. For these signals to be displayed, the monitor must be manually reconfigured away from the default state. However, the displaying of the pacemaker impulse remains a digital process, and the monitor "paints" this signal (and other noise) back into the displayed ECG as a pacemaker impulse. Two surgical cases are reported that demonstrate monitor malfunction in systems with digitally processed ECGs during application of nerve stimulator signals to the patient. In one case, application of a 100-Hz tetany signal completely abolished the displayed pacemaker pulses. In the second case, application of a 50-Hz tetany signal abolished appropriate display but caused the monitor to display artifactual pacemaker pulses. Knowledge of undesirable monitor behavior can reduce inappropriate therapy in the operating room. PMID- 15110374 TI - Ultrasound-guided percutaneous dilatational tracheostomy with laryngeal mask airway control in a morbidly obese patient. AB - Percutaneous dilatational tracheostomy (PDT) is a widely used and accepted method of long-term ventilation of critically ill patients in many intensive care units. However, it has certain contraindications that must be taken into account; for example, difficult anatomy and short, bull neck that are so often seen in morbidly obese persons. We present a case of a morbidly obese female patient in whom ultrasound-guided PDT was performed and in whom the airway was controlled by Laryngeal Mask Airway (LMA) during the procedure. Possible advantages of an ultrasonography-guided method and LMA control in morbidly obese patients also are discussed. PMID- 15110375 TI - Dexmedetomidine infusion for sedation during fiberoptic intubation: a report of three cases. AB - We report three patients undergoing cervical spine surgery who required awake fiberoptic intubation, and in whom sedation was provided using a dexmedetomidine infusion. Dexmedetomidine was used to provide a moderate level of sedation without causing respiratory distress or hemodynamic instability during fiberoptic intubation. Conditions for intubation were acceptable in all three patients after co-administration of topical anesthesia. Dexmedetomidine may serve as a useful adjunct for this procedure. The anesthetic management and anesthetic implications of using dexmedetomidine infusions for awake fiberoptic intubation are discussed. PMID- 15110376 TI - Carotid-jugular arteriovenous fistula: case report of an iatrogenic complication following internal jugular vein catheterization. AB - Central venous catheterization is frequently performed for perioperative management and long-term intravenous access. Although the complications of central venous catheter insertion have been widely reported, there are few reports of carotid-jugular arteriovenous fistula formation. We describe a case of a carotid-jugular arteriovenous fistula following the insertion of a double-lumen catheter for hemodialysis access. We provide recommendations for the prevention and early detection of this iatrogenic complication. PMID- 15110377 TI - Anesthesiology: perioperative medicine or "when the anesthetic is a diuretic". AB - Two patients are reported, each with heart failure, who were treated with digoxin (case 1) and furosemide (case 2). Indications for medical treatment of patients with heart failure, the role of various drugs, and exercise therapy, are reviewed. At a time when the population of people over 65 years of age is increasing, it is important for physicians to recognize the symptoms of heart failure and to know the most up-to-date treatment for this disorder. These cases demonstrate the significance of the anesthesiologist as a perioperative physician. PMID- 15110378 TI - A case of status asthmaticus complicated by anginal pain and an elevation in blood cpk levels during tapering of isoproterenol administered for management of respiratory failure. PMID- 15110379 TI - Patient satisfaction and outcome in thyroidectomy and parathyroidectomy in patients receiving propofol. PMID- 15110380 TI - The reach of patient simulation: is there no boundary? PMID- 15110381 TI - Using high-fidelity patient simulation and an advanced distance education network to teach pharmacology to second-year medical students. AB - The feasibility and acceptance of an Advanced Distance Education Network (ADEN) in bringing the simulated operating room (OR) to second-year medical students learning the pharmacology of anesthetic drugs is reviewed. A MedSim-Eagle (Binghamton, NY) full-scale mannequin simulator was used. Using an ADEN, students were linked in real time to a simulated OR where the anesthesiologist instructor was using a MedSim-Eagle patient simulator to present for discussion the physiologic effects of volatile anesthetics on cardiac output (CO), heart rate (HR), mean arterial pressure (MAP), and systemic vascular resistance (SVR). The use of simulation to present basic science principles of isoflurane and halothane's effect on CO, HR, MAP, and SVR in a clinical setting via an ADEN is feasible. Student acceptance of this method of education is high, as measured by a post-exercise survey. Ninety-five percent of students felt this exercise was a valuable use of their time; 93% felt the ADEN-delivered clinical simulation presentation contributed to their understanding of the pharmacology of anesthesia. Eighty-three percent of students preferred this integrated clinically oriented review to a didactic review of the material, and 92% of students who had experienced previous small group hands-on session simulation felt the ADEN delivered session was the same or better. PMID- 15110382 TI - A multidisciplinary approach to teach responses to weapons of mass destruction and terrorism using combined simulation modalities. AB - STUDY OBJECTIVE: To reinforce concepts presented in the lectures; understand the complexity and speed of casualty and information generation during a Weapons of Mass Destruction and Terrorism (WMD/T) event; experience the novelty of combined weapons' effects; recognize the time course of the various chemical, biological, and radiation agents; and make challenging decisions with incomplete and conflicting information. SETTINGS: Two environments simulated simultaneously: one a major trauma center emergency room (ER) with two patient simulators and several human actors; the other an Emergency Operations Command Center (EOC). TARGET AUDIENCE: Students for this course included: clinicians, scientists, military and intelligence officers, lawyers, administrators, and logistic personnel whose jobs involve planning and executing emergency response plans to WMD/T. SIMULATION SCRIPT: A WMD/T attack in Washington, D.C., has occurred. Clinical students performed in their real life roles in the simulated ER, while nonclinical students did the same in the simulated EOC. Six ER casualties with combined WMD/T injuries were presented and treated over 40 minutes. In the EOC, each person was given his or her role title with identification tag. The EOC scenario took cues from the action in the ER via two television (TV) news feeds and telephone calls from other Emergency Operations Assets. PERFORMANCE EXPECTATIONS: Students were expected to actively engage in their roles. Student performances were self evaluated during the debriefing. DEBRIEFING: The two groups were reunited and debriefed utilizing disaster crisis resource management tools. ASSESSMENT OF EFFECTIVENESS: Students answered an 18-point questionnaire to help evaluate the usefulness and acceptance of multimodality patient simulation. LESSONS LEARNED: Large-scale multimodality patient simulation can be used to train both clinicians and nonclinicians for future events of WMD/T. Students accepted the simulation experience and thought that scenario was appropriately realistic, complex, and overwhelming. Difficulties include the extensive man-hours involved in designing and presenting the live simulations. EOC-only sessions could be staged with only a few video cassette recorders, TVs, telephones, and callers. PMID- 15110383 TI - Introduction to serial reviews on EpRE and its signaling pathway. PMID- 15110384 TI - Nrf2 signaling in coordinated activation of antioxidant gene expression. AB - Antioxidant response element (ARE)-mediated expression and coordinated induction of antioxidant enzymes is a critical mechanism of protection against chemically induced oxidative/electrophilic stress. NF-E2-related nuclear factors (Nrf1 and Nrf2) bind to ARE and regulate ARE-mediated gene expression and induction. Nrf2 is more potent than Nrf1 in activation of ARE-regulated gene expression. Nrf2 is retained in the cytoplasm by an inhibitor INrf2. Nrf2 binding to INrf2 leads to proteasomal degradation of Nrf2. An increase in oxidative/electrophilic stress, due to chemical exposure, leads to the activation of protein kinase C (PKC) and other cytosolic factors. PKC phosphorylation of Nrf2 at serine 40 results in the escape or release of Nrf2 from INrf2. Nrf2 translocates to the nucleus, forms heterodimers with its unknown partner proteins, and binds to the ARE. This leads to the coordinated activation of ARE-regulated genes. Additional nuclear factor including small Mafs (MafG and MafK), large Maf (c-Maf), c-Fos, and Fra1, also bind to ARE and negatively regulate ARE-mediated gene expression. This is presumably to keep the expression of antioxidant enzymes "in check" to maintain the cellular defenses active and/or to rapidly restore induced enzymes to normal levels. Future investigations are expected to reveal that a balance between positive and negative factors regulates ARE-mediated gene expression and induction. The future studies should also reveal a complete mechanism of signal transduction from antioxidants and xenobiotics to the transcription factors, such as Nrf2, that bind to ARE. PMID- 15110385 TI - Molecular mechanism activating Nrf2-Keap1 pathway in regulation of adaptive response to electrophiles. AB - Electrophile responsive element (EpRE)-mediated gene induction is a pivotal mechanism of cellular defense against the toxicity of electrophiles and reactive oxygen species (ROS). Nrf2, which belongs to the cap'-n'-collar family of basic region-leucine zipper transcription factors, has emerged as an essential component of an EpRE-binding transcriptional complex. Detailed analysis of the regulatory mechanism governing Nrf2 activity led to the identification of Keap1, which represses Nrf2 activity by directly binding to the N-terminal Neh2 domain. Keap1 interaction with Neh2 leads to the sequestration of Nrf2 in the cytoplasm and to the enhancement of Nrf2 degradation by proteasomes conferring tight regulation on the response. Electrophiles act to counteract sequestration of Nrf2 by Keap1 and provoke Nrf2 activation. Constitutive activation of Nrf2-regulated transcription in Keap1 knockout mice clearly demonstrated that the disruption of Keap1 repression is sufficient for the activation of Nrf2. These observations indicated that the mechanism that modulates Nrf2-Keap1 interaction is pivotal for the cellular sensing mechanism for electrophiles. Recent analyses argue that the redox mechanism that modifies cysteine residues of Keap1 governs the Keap1-Nrf2 interaction and therefore is critical for sensing of electrophiles. PMID- 15110386 TI - Using anti-5,5-dimethyl-1-pyrroline N-oxide (anti-DMPO) to detect protein radicals in time and space with immuno-spin trapping. AB - The detection of protein free radicals using the specific free radical reactivity of nitrone spin traps in conjunction with nitrone-antibody sensitivity and specificity greatly expands the utility of the spin trapping technique, which is no longer dependent on the quantum mechanical electron spin resonance (ESR). The specificity of the reactions of nitrone spin traps with free radicals has already made spin trapping with ESR detection the most universal, specific tool for the detection of free radicals in biological systems. Now the development of an immunoassay for the nitrone adducts of protein radicals brings the power of immunological techniques to bear on free radical biology. Polyclonal antibodies have now been developed that bind to protein adducts of the nitrone spin trap 5,5 dimethyl-1-pyrroline N-oxide (DMPO). In initial studies, anti-DMPO was used to detect DMPO protein adducts produced on myoglobin and hemoglobin resulting from self-peroxidation by H2O2. These investigations demonstrated that myoglobin forms the predominant detectable protein radical in rat heart supernatant, and hemoglobin radicals form inside red blood cells. In time, all of the immunological techniques based on antibody-nitrone binding should become available for free radical detection in a wide variety of biological systems. PMID- 15110387 TI - Spin trapping of glutathiyl and protein radicals produced from nitric oxide derived oxidants. AB - Despite the importance of protein radicals in cell homeostasis and cell injury, their formation, localization, and propagation reactions remain obscure, mainly because of the difficulties in detecting and characterizing radicals, in general, and protein radicals, in particular. New approaches based on spin trapping coupled with other methodologies are under development/testing but so far they have been applied mainly to the study of protein-tyrosyl and protein-tryptophanyl radicals. Here, our aim is to emphasize the importance of developing new methodologies for the detection of glutathyil and protein-cysteinyl radicals under physiological conditions. To this end, we summarize current EPR evidence supporting the view that glutathione and protein-cysteines are among the preferential targets of nitric oxide-derived oxidants and that they are oxidized to the glutathiyl and protein-cysteinyl radicals, respectively. The possible intermediacy of these species in the biological formation of mediators of protein cysteine redox signaling, such as S-nitrosothiols and sulfenic acids, is also discussed. PMID- 15110388 TI - Dopaminergic modulation of oxidative stress and apoptosis in human peripheral blood lymphocytes: evidence for a D1-like receptor-dependent protective effect. AB - Dopamine (DA) is a neurotransmitter in the central and peripheral nervous system, which can be either cytotoxic or cytoprotective under selected conditions. Such effects involve oxidative mechanisms and are likely to play a role in neurodegenerative disorders. Because increasing evidence points to peripheral blood lymphocytes (PBL) as a feasible model for studying DA-related mechanisms of cell death and survival, we have explored in these cells the effects of DA on oxidative metabolism and apoptosis. Our results show that, whereas DA 100-500 microM resulted in increased intracellular reactive oxygen species (ROS) levels and apoptotic cell death through oxidative stress, DA 0.1-5 microM decreased ROS levels and apoptosis. DA (both 1 and 500 microM) partially counteracted the decrease in Cu/Zn superoxide dismutase levels observed in untreated PBL. However, whereas the effect of the low dose lasted for the whole incubation period (24 h), the effect of DA 500 microM was transient. DA-dependent reduction of ROS levels and apoptosis was prevented by D1-like (but not D2-like) receptor antagonism. The present findings add knowledge about the sensitivity of PBL to DA and strengthen the rationale for exploiting these cells as an easily accessible peripheral model for the ex vivo investigation of oxidative stress-related dopaminergic mechanisms underlying human neurodegenerative diseases. PMID- 15110389 TI - N epsilon-carboxymethyllysine in brain aging, diabetes mellitus, and Alzheimer's disease. AB - Oxidative stress has been implicated in the pathophysiology of Alzheimer's disease (AD) and diabetes mellitus (DM). N epsilon-carboxymethyllysine (CML) is an advanced glycation end product (AGE) recently found to be associated with oxidative stress mechanisms. Using immunocytochemical methods we examined the distribution of CML in brain tissue from AD and DM subjects and aging controls. CML reactivity was present in the cytoplasm of neurons, but there were marked differences in the intensity of expression, number of cells, and topographical distribution. CML expression was higher in hippocampus than in frontal and temporal cortex. In the hippocampus, neuronal and, to an extent, glial expression was more marked in CA3 and CA4 than in CA1 and CA2. In AD, CML was found to be coexpressed with tau protein, showing the similar neurofibrillary tangle shape, as well as in neuritic plaques but not in the core of amyloid plaques. We noted an increasing degree of CML expression such that the highest reactivity was evident in those with both AD and DM, followed by AD, DM, and aging controls. There was an inverse relationship between Braak staging and topography of CML expression. Although DM cases did not show Abeta deposition or neurofibrillary tangles, these findings suggest increased CML expression is not limited to AD. Nonetheless, high CML expression in AD with coexistent DM suggests there are additive effects compared with AD alone. It is plausible that the microangiopathy also associated with DM could worsen AD pathogenesis. PMID- 15110390 TI - Polyphenols from Camellia sinenesis prevent primary graft failure after transplantation of ethanol-induced fatty livers from rats. AB - Fatty liver caused by ethanol decreases survival after liver transplantation in rats. This study investigated if antioxidant polyphenols from Camellia sinenesis (green tea) prevent failure of fatty grafts from ethanol-treated rats. Donor rats were given ethanol intragastrically (6 g/kg). After 20 h, livers were explanted and stored in University of Wisconsin solution for 24 h. Prior to implantation, the explanted grafts were rinsed with lactated Ringer's solution containing 0 to 60 microg/ml polyphenols. Alanine aminotransferase (ALT) release after liver transplantation was 4.5-fold higher in recipients receiving ethanol-induced fatty grafts than in those receiving normal grafts. Liver grafts from ethanol-treated donors also developed severe focal necrosis. Graft survival was 11% in the ethanol group versus 88% for normal grafts. Polyphenol treatment at 60 microg/ml blunted ALT release by 66%, decreased necrotic areas by 84%, and increased survival to 75%. Ethanol increased alpha-(4-pyridyl-1-oxide)-N-tert.-butylnitrone free radical adducts in bile by 2.5-fold, as measured by electron spin resonance spectroscopy, and caused accumulation of 4-hydroxynonenal in liver sections, effects blunted by polyphenols. Epicatechin gallate, a major polyphenol from C. sinenesis, also decreased enzyme release, minimized pathological changes, and decreased free radical adduct formation. In conclusion, polyphenols scavenged free radicals in ethanol-induced fatty livers and decreased injury after liver transplantation. PMID- 15110391 TI - p38 MAPK associated with stereoselective priming by grepafloxacin on O2- production in neutrophils. AB - Grepafloxacin is an asymmetric fluoroquinolone derivative which possesses high tissue penetrability as well as strong, broad-spectrum antimicrobial activities. We recently found that grepafloxacin induced a priming effect on neutrophil respiratory burst induced by N-formylmethionylleucylphenylalanine. In this report, we elucidate the precise mechanism of the priming by grepafloxacin. The R(+) enantiomer of grepafloxacin induced a more potent priming effect than did S( )-grepafloxacin. R(+)-Grepafloxacin also produced a more potent translocation of both p47- and p67-phox proteins to membrane fractions of neutrophils. Grepafloxacin-induced primed superoxide generation was significantly inhibited by pretreatment with PD169316 and SB203580, p38 mitogen-activated protein kinase (MAPK) inhibitors, but not with PD98059, a specific inhibitor of the upstream kinase that activates p44/42 MAPK, or SP600125, an inhibitor of stress-activated protein kinase/c-Jun N-terminal kinase (JNK). Grepafloxacin strongly phosphorylated p38 MAP kinase but not p44/42 MAPK or JNK. R(+)-Grepafloxacin showed more potent phosphorylation of p38 MAPK than did S(-)-grepafloxacin, in a time- and concentration-dependent manner. PD169316 significantly inhibited R(+) grepafloxacin-induced translocation of p47-phox protein to the membrane fraction. Interestingly, grepafloxacin stereospecifically bound to the membrane fractions of neutrophils. These results strongly suggest that grepafloxacin stereospecifically primes neutrophil respiratory burst, and p38 MAPK activation is closely related to the grepafloxacin priming. PMID- 15110392 TI - Human mitochondrial thioredoxin reductase reduces cytochrome c and confers resistance to complex III inhibition. AB - The ubiquitously expressed mammalian thioredoxin reductases are selenoproteins that together with NADPH regenerate active reduced thioredoxins and are involved in diverse actions mediated by redox control. Two main forms of mammalian thioredoxin reductases have been isolated, one cytosolic (TrxR1) and one present in mitochondria (TrxR2). Although the principal target for TrxRs is thioredoxin, the cytosolic form can regenerate several important antioxidants such as ascorbic acid, lipoic acid, and ubiquinone. In this study we demonstrate that cytochrome c is a substrate for both TrxR1 and TrxR2. In addition, cells overexpressing TrxR2 are more resistant to impairment of complex III in the mitochondrial respiratory chain upon both antimycin A and myxothiazol treatments, suggesting a complex III bypassing function of TrxR2. Furthermore, we show that cytochrome c is reduced by TrxR2 in vitro, not only by using NADPH as an electron donor but also by using NADH, pointing at TrxR2 as an important redox protein on complex III impairment. These findings may be valuable in understanding respiratory disorders in mitochondrial diseases. PMID- 15110393 TI - Upregulation of NAD(P)H oxidase 1 in hypoxia activates hypoxia-inducible factor 1 via increase in reactive oxygen species. AB - Hypoxia sensing and related signaling events, including activation of hypoxia inducible factor 1 (HIF-1), represent key features in cell physiology and lung function. Using cultured A549 cells, we investigated the role of NAD(P)H oxidase 1 (Nox1), suggested to be a subunit of a low-output NAD(P)H oxidase complex, in hypoxia signaling. Nox1 expression was detected on both the mRNA and protein levels. Upregulation of Nox1 mRNA and protein occurred during hypoxia, accompanied by enhanced reactive oxygen species (ROS) generation. A549 cells, which were transfected with a Nox1 expression vector, revealed an increase in ROS generation accompanied by activation of HIF-1-dependent target gene expression (heme oxygenase 1 mRNA, hypoxia-responsive-element reporter gene activity). In A549 cells stably overexpressing Nox1, accumulation of HIF-1alpha in normoxia and an additional increase in hypoxia were noted. Interference with ROS metabolism by the flavoprotein inhibitor diphenylene iodonium (DPI) and catalase inhibited HIF 1 induction. This suggests that H2O2 links Nox1 and HIF-1 activation. We conclude that hypoxic upregulation of Nox1 and subsequently augmented ROS generation may activate HIF-1-dependent pathways. PMID- 15110394 TI - Validation of Trypanosoma brucei trypanothione synthetase as drug target. AB - In trypanosomes, the parasite-specific thiol trypanothione [T(SH)2] fulfills various functions, the best established being detoxification of H2O2 and organic hydroperoxides and ribonucleotide reduction. Recently, a trypanothione synthetase (Tb-TryS) gene from Trypanosoma brucei was isolated and the heterologously expressed Tb-TryS catalyzed the entire synthesis of T(SH)2 from glutathione (GSH) and spermidine in vitro. To confirm the in situ function of the complex Tb-TryS activities and to evaluate the importance of T(SH)2 metabolism in T. brucei, TryS suppression by double-stranded RNA interference was performed. Knockdown of TryS led to depletion of both T(SH)2 and glutathionylspermidine (Gsp) and accumulation of GSH, while concomitantly impairment of viability and arrest of proliferation were observed. TryS-downregulated cells displayed a significantly increased sensitivity to H2O2 and tert.-butyl hydroperoxide. These data verify the hypothesis that in T. brucei, a single enzyme synthesizes the spermidine conjugated thiols (Gsp and T(SH)2) and further confirms the significance of trypanothione in the defense against oxidative stress and the maintenance of viability and proliferation in unstressed parasites. PMID- 15110395 TI - Antioxidant properties of S-adenosyl-L-methionine in Fe(2+)-initiated oxidations. AB - S-Adenosylmethionine (SAM) is protective against a variety of toxic agents that promote oxidative stress. One mechanism for this protective effect of SAM is increased synthesis of glutathione. We evaluated whether SAM is protective via possible antioxidant-like activities. Aerobic Hepes-buffered solutions of Fe2+ spontaneously oxidize and consume O2 with concomitant production of reactive oxygen species and oxidation of substrates to radical products, e.g., ethanol to hydroxyethyl radical. SAM inhibited this oxidation of ethanol and inhibited aerobic Fe2+ oxidation and consumption of O2. SAM did not regenerate Fe2+ from Fe3+ and was not consumed after incubation with Fe2+. SAM less effectively inhibited aerobic Fe2+ oxidation in the presence of competing chelating agents such as EDTA, citrate, and ADP. The effects of SAM were mimicked by S adenosylhomocysteine, but not by methionine or methylthioadenosine. SAM did not inhibit Fe2+ oxidation by H2O2 and was a relatively poor inhibitor of the Fenton reaction. Lipid peroxidation initiated by Fe2+ in liposomes was associated with Fe2+ oxidation; these two processes were inhibited by SAM. However, SAM did not show significant peroxyl radical scavenging activity. SAM also inhibited the nonenzymatic lipid peroxidation initiated by Fe2+ + ascorbate in rat liver microsomes. These results suggest that SAM inhibits alcohol and lipid oxidation mainly by Fe2+ chelation and inhibition of Fe2+ autoxidation. This could represent an important mechanism by which SAM exerts cellular protective actions and reduces oxidative stress in biological systems. PMID- 15110396 TI - Nitric oxide and BCNU chemoresistance in C6 glioma cells: role of S nitrosoglutathione. AB - Inducible nitric oxide synthase (iNOS or NOS2) is expressed in malignant glioma. Previously we noted that C6 glioma cells overexpressing NOS2 displayed chemoresistance against 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU) and other chloroethylnitrosourea derivatives with carbamoylating action. Herein we report experimental evidence supporting the contention that this NOS2 effect is mediated, at least in part, by S-nitrosoglutathione (GSNO), a potent antioxidant derived from interaction of NO and glutathione. Out of three NO donors tested, only GSNO was effective in protecting glioma cells against BCNU cytotoxicity. Furthermore, the protective effect of GSNO, similar to that of NOS2, was confined to carbamoylating, but not alkylating action. Experimental manipulations that were expected to increase or decrease cellular GSNO stores, as confirmed by immunocytochemical staining using a GSNO-specific antibody and HPLC analysis of GSNO contents in culture medium, led respectively to enhanced or reduced chemoresistance against carbamoylating cytotoxicity. Finally, neocuproine, a selective cuprous ion chelator known to neutralize GSNO actions, abolished NOS2 mediated chemoresistance against carbamoylating agents. Our results reveal a novel action of NOS2/GSNO that may potentially contribute to the development of chemoresistance against BCNU, which remains a mainstay in chemotherapy for glioblastoma multiforme. PMID- 15110397 TI - Antioxidant supplementation prevents exercise-induced lipid peroxidation, but not inflammation, in ultramarathon runners. AB - To determine if 6 weeks of supplementation with vitamins E and C could alleviate exercise-induced lipid peroxidation and inflammation, we studied 22 runners during a 50 km ultramarathon. Subjects were randomly assigned to one of two groups: (1) placebos (PL) or (2) antioxidants (AO: 1000 mg vitamin C and 300 mg RRR-alpha-tocopheryl acetate). Blood samples were obtained prior to supplementation (baseline), after 3 weeks of supplementation, 1 h pre-, mid-, and postrace, 2 h postrace and for 6 days postrace. Plasma levels of alpha-tocopherol (alpha-TOH), ascorbic acid (AA), uric acid (UA), F2-isoprostanes (F2-IsoPs), tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and C-reactive protein (CRP) were measured. With supplementation, plasma alpha-TOH and AA increased in the AO but not the PL group. Although F2-IsoP levels were similar between groups at baseline, 28 +/- 2 (PL) and 27 +/- 3 pg/ml (AO), F2-IsoPs increased during the run only in the PL group (41 +/- 3 pg/ml). In PL women, F2 IsoPs were elevated postrace (p <.01), but returned to prerace concentrations by 2 h postrace. In PL men, F2-IsoP concentrations were higher postrace, 2 h postrace, and 1, 2, 3, 4, and 6 days postrace (PL vs. AO group, each p <.03). Markers of inflammation were increased dramatically in response to the run regardless of treatment group. Thus, AO supplementation prevented endurance exercise-induced lipid peroxidation but had no effect on inflammatory markers. PMID- 15110398 TI - Coccidiosis control by administering toltrazuril in the drinking water for a 2 day period. AB - A 56-day floor pen study was conducted to determine the appropriate time to administer toltrazuril (Baycox) (TOL) for control of coccidiosis in broiler chickens. Litter was seeded with field strains of Eimeria acervulina, Eimeria maxima and Eimeria tenella. On Days 0, 21, 35 and 56, all birds and feed were weighed. Starting on Day 14, weekly lesion scores and oocyst counts were performed. The treatments were 125 ppm nicarb (NIC) in the starter to 66 ppm salinomycin (SAL) in the grower with no TOL (NIC/SAL/no TOL), 66 ppm salinomycin in both the starter and the grower but no TOL (SAL/SAL/no TOL), or no in-feed medication with the following TOL treatment: TOL Days 2-3, TOL Days 6-7, TOL Days 10-11, TOL Days 14-15, TOL Days 18-19, and as control NM/NM/no TOL (NM). The withdrawal feed was nonmedicated. TOL was administered in the drinking water at the rate of 7 mg/kg body weight. Oocysts per gram litter and lesion scores showed a significant infection in the NM birds, which peaked about Day 21. The NIC/SAL gave excellent early protection but only moderate protection during the SAL phase. The final performance for the SAL/SAL was significantly less compared to all TOL and NIC/SAL birds. All TOL treatments but Days 2-3 provided good coccidiosis control with accompanying performance. The absence of clinical coccidiosis relapse during the last third of the growout along with moderate oocyst counts and low lesions was indicative of unimpaired coccidiosis immunity. It can be inferred from the overall results that the use of TOL as the sole anticoccidial for two consecutive days in the drinking water between Days 10 and 14 would be the best time for good coccidiosis control allowing full performance. PMID- 15110399 TI - Prevalence of intestinal parasites in dogs under veterinary care in Maracaibo, Venezuela. AB - The prevalence of intestinal parasites in dogs presented to the Veterinary Policlinic of the University of Zulia (PVU) was measured between January and December 2001. A total of 614 fecal samples were evaluated by the fecal flotation method. One or more species of parasites was identified in 218 (35.5%) dogs. The parasites most frequently detected were: Ancylostoma spp. (24.5%), Toxocara canis (11.4%) and Isospora spp. (8.1%). Single parasitic infections were present in 149 (24.3%) dogs. The age distribution of intestinal parasites in dogs less than 1 year old had a higher overall prevalence than those dogs over 12 months of age. There was no significant difference in the prevalence between male (38.9%) and female (31.7%) dogs. There was a significantly (P < 0.05) greater prevalence of parasites in mixed-breed dogs (40.3%) as compared with pure-breed dogs (30.8%). A significant difference (P < 0.05) was detected between the general prevalence of January and December compared to August. PMID- 15110400 TI - Sero-epidemiological study of canine Leishmania spp. infection in the municipality of Alijo (Alto Douro, Portugal). AB - Visceral leishmaniosis caused by Leishmania infantum is a zoonotic disease in the Mediterranean basin. We report an epidemiological survey carried out in dogs from the municipality of Alijo in the endemic region of Alto Douro (north Portugal). Performance of the direct agglutination test (DAT) was assessed in 205 matching samples of blood collected on filter paper and serum. A high degree of agreement (97.6%; k = 0.83) was found between the results obtained from both types of samples. DAT was then used to test more blood on filter paper (B-FP) samples from other dogs of the same municipality. The detected sero-prevalence was 18.7% (288/1540), with values ranging from 0.0 to 81.1% in each of the 19 parishes of Alijo. Three distinct geographical zones of mean sero-prevalence could be defined: northwestern (2.5%), intermediate (11.4%) and southern (49.9%). No statistically significant difference was observed between male (19.1%) and female (17.8%) sero-prevalences (P = 0.560). Dogs of 9-11 years of age showed the highest sero-prevalence (28.4%), but all the other age-intervals (0-2, 3-5, 6-8 and 12-17 years) presented values (15.0-22.3%) not significantly different from the mean of the whole study population. Risk factors for canine Leishmania infection were age and geographical zone. Only 5.9% of the sero-positive animals had clinical signs of canine leishmaniosis and the overall prevalence of disease was 1.1%. This study validates the use of B-FP samples and confirms DAT as a simple and sensitive serological test to evaluate the level of canine Leishmania infection in areas of high sero-prevalence. PMID- 15110401 TI - Evaluation of ovine abortion associated with Toxoplasma gondii in Spain by different diagnostic techniques. AB - A total of 173 aborted ovine foetuses and seven aborted caprine foetuses, submitted from different points of north and central Spain, were analysed to determine the role of T. gondii in abortion and to compare the utility of the most widely used techniques in diagnosis of the congenital infection (histopathology, serology--IFAT and ELISA--and a nested-PCR). Parasite infection was diagnosed in 40 (23.1%; n = 173) ovine foetuses by at least one of the diagnostic techniques used. A higher percentage of foetuses were diagnosed using serological techniques (IFAT and ELISA) (28.3%; n = 106) than by histologic examination (8.7%; n = 173) or PCR (6.9%; n = 173). No significant association between infection and the foetal age categories was found (P > 0.05). In this study, 106 aborted foetuses were analysed by all of the three diagnostic techniques. When we compared serological results, perfect agreement between ELISA and IFAT was obtained. On the contrary, slight to fair agreements were observed when histology results were compared with those obtained by serology and PCR techniques. All the positive foetuses were aborted in the mid (60%) or last (40%) term of pregnancy, but no significant differences were found between ages of the infected and non-infected foetuses (P > 0.05). This report indicates that toxoplasmosis may be a common cause of small ruminant abortion and neonatal death in Spain and points out the necessity of using different and complementary techniques to increase the probability of detecting Toxoplasma infection in an aborted foetus. PMID- 15110402 TI - Evaluation of the card agglutination test (CATT/T. evansi) for detection of Trypanosoma evansi infection in water buffaloes (Bubalus bubalis) in Egypt. AB - A card agglutination test (CATT/T. evansi) was evaluated for detection of antibodies against Trypanosoma evansi (T. evansi) in experimentally and naturally infected buffaloes. Four calves were inoculated with a strain of T. evansi isolated from a dromedary camel. Parasitological examination of the calves revealed trypanosomes in the blood from days 4 to 9 post-inoculation (PI). General emaciation appeared from day 26 PI and aggravated until the end of the experiment (day 88 PI). Antibodies against T. evansi were detectable from day 8 PI till the end of the experiment. Parasitological examination of 200 water buffalo blood samples obtained from slaughterhouses revealed negative results. Serological examination of these animals showed that 48 (24%) water buffaloes had anti-T. evansi antibodies. PMID- 15110404 TI - A membrane-anchored Theileria parva cyclophilin with a non-cleaved amino-terminal signal peptide for entry into the endoplasmic reticulum. AB - Recent studies suggest that peptidyl-prolyl isomerases of the cyclophilin family, that access the secretory pathway, can be involved in the interaction of parasitic protozoa with mammalian host cells. The amino acid sequence of a cDNA encoding a cyclophilin family member of the intracellular protozoan parasite of cattle Theileria parva contains a conserved C-terminal domain that exhibits 70% amino acid identity to cyclophilin proteins from other organisms, and a unique 60 amino acid novel N-terminal extension. Cell-free expression of the cDNA revealed a 26kDa amino translation product, indicating expression of the N-terminal domain. The protein-coding region contains three short introns, less than 100 base pairs in length and Northern blot analysis demonstrates expression of a single 0.9 kb transcript in the piroplasm and schizont stages. The transcript is present in high abundance in the intra-lymphocytic schizont stage. The recombinant protein binds to immobilized cyclosporin A, a finding consistent with peptidyl-prolyl cis-trans isomerase function in vivo. A predicted N-terminal signal peptide was functional for entry into the eukaryotic secretory transport pathway in a cell-free in vitro transcription/translation system. The C-terminal cyclophilin domain was translocated across the membrane of the endoplasmic reticulum and the uncleaved signal peptide functioned as a membrane anchor. PMID- 15110403 TI - Improved immunogenicity of novel baculovirus-derived Theileria parva p67 subunit antigens. AB - East Coast fever (ECF) in cattle is caused by the tick-borne protozoan parasite Theileria parva. The major sporozoite surface antigen of T. parva (p67) is an important candidate for inclusion in a subunit vaccine. Recently, we reported the expression and production of different parts of p67 as fusions to either GFP or to the baculovirus GP64 envelope glycoprotein in insect cells, which resulted in stable proteins recognized by a monoclonal specific for native p67. The immunogenicity of these fusion proteins was examined in out-bred mice and cattle. In mice, the full length p67 molecule without its signal peptide and transmembrane region, but fused to GFP (GFP:p67deltaSS) was the best immunogen followed by the C-terminus of p67 fused to GP64 (GP64:p67C). These two immunogens also provoked a high level of sero-conversion in cattle when formulated in a water-in-oil or saponin-derived adjuvant with only 100 microg of protein and a single booster. The vaccine-elicited antibodies efficiently inhibited the infectivity of T. parva sporozoites in in vitro neutralization assays. This study demonstrated that these new baculovirus-derived p67 vaccines were highly immunogenic, and that in combination with a suitable adjuvant, they have a clear potential to induce protective immunity in cattle. PMID- 15110405 TI - Recirculation of Trypanosoma brucei brucei in cattle after T. congolense challenge by tsetse flies. AB - The effect of challenging cattle, chronically infected with Trypanosoma brucei brucei, with T. congolense on the development of the T. b. brucei infection was investigated. For this purpose, nine experimental animals were first infected with T. b. brucei through the bites of infected tsetse flies. Once the T. b. brucei had developed into a chronic infection, that was difficult to detect using routine parasitological diagnostic tools, seven of the experimental animals were challenged by tsetse flies infected with T. congolense. Two of the animals infected with T. b. brucei were kept as control. The infection with T. congolense resulted in a sudden increase in the parasitaemia of T. b. brucei. In the T. b. brucei control animals, on the other hand, the parasitaemia remained below the level of detection. The epidemiological repercussions of this increase in the parasitaemia of T. b. brucei after infection with T. congolense are discussed. PMID- 15110406 TI - Dynamics of Fasciola hepatica infection in two species of snails in a rural locality of Mexico. AB - The aim of the present study was to identify the species that are natural intermediate hosts of Fasciola hepatica in a Research Centre in Chapa de Mota, State of Mexico, Mexico, where temperate and humid conditions prevail. Also, the magnitude and dynamics of the natural infection in four biotopes were analysed. The study was performed between October 2000 and November 2001. The molluscs were collected, identified and studied for trematode infection. Soil humidity and temperature were determined monthly. A total of 4042 snails were collected 3372 (83%) were Fossaria humilis and 670 (17%) F. bulimoides. F. hepatica was present in 2537 (75.2%) and 515 (76.9%) of the two snail species, respectively. Temperature, soil humidity and microenvironment affected the dynamics of the host population and the parasite infestation, with a significant increase from August to November 2001. The highest prevalence was observed in the warmer and more humid biotopes. PMID- 15110407 TI - Genetic relationships between sheep, cattle and human Echinococcus infection in Tunisia. AB - Allozyme variation at seven polymorphic loci (GPI, EST, MDH, MPI, DIA, PEP, PGM) was studied to examine genetic variation within and between sheep, cattle and human populations of Echinococcus granulosus in Tunisia. A high degree of genetic similarity was shown between the cysts of the three host origins. Nevertheless, whereas, the ovine and human samples were highly similar, the cattle samples were slightly different genetically. We conclude that humans are mostly infected by parasites originating from sheep liver. The intense deficiency in heterozygotes was partly artefactual (Wahlund effect) and partly due to self-fertilisation. PMID- 15110408 TI - Faecal egg count reduction test for assessing anthelmintic efficacy: average versus individually based estimations. AB - The faecal egg count reduction (FECR) in faeces is the most widely used method to assess the efficacy of anthelmintics against gastrointestinal strongyles. Several methods are used and they yield different efficacy values. They give important weight to those animals which excrete the majority of eggs, before or after treatment. We propose using individual animal FECR tests that give equal weight to every tested host. The first method is based on a before and after treatment without untreated controls, whereas the second one takes into account the use of non-treated control hosts. Confidence interval at P = 0.95 were established on bootstrap resamples for both individual-based methods. The individual-based FECR presented lower values than the average-based FECR in most cases. The individual FECR provided reliable evaluation when egg counts were over 300 eggs/g and when at least 10 animals were tested. PMID- 15110409 TI - Zoonotic risk of Toxocara canis infection through consumption of pig or poultry viscera. AB - The potential zoonotic risk of Toxocara canis infections from consumption of swine or poultry viscera containing larvae was assessed using a pig model. Two groups of six pigs were fed either fresh swine viscera (group FS) or poultry viscera (FP) containing around 3500 Toxocara larvae. Another two groups of six pigs were fed swine viscera (PS) or poultry viscera (PP) preserved at 4 degrees C for 1 week. All pigs were necropsied 14 days after the exposure. Liver white spots were counted and T. canis specific IgG antibodies were measured by ELISA. Larval burdens were assessed in the mesenteric lymph nodes, liver, lungs, brain, tongue, and eyes. All recipient pigs exhibited several white spots on the liver surface and detectable antibody levels. Larvae were recovered predominantly from the lungs, but also from the mesenteric lymph nodes and the liver, a few larvae were found in the brain and tongue of the pigs. Two larvae were found in the eyes of two pigs in group FS. Mean percentages of total larval recoveries in groups FS, FP, PS, and PP were 75.3, 63.6, 42.6, and 18.8%, respectively. Significantly higher numbers of larvae were recovered from pigs given swine viscera than pigs given poultry viscera. The preservation at 4 degrees C for 1 week caused a significant reduction in the larval infectivity overall, nevertheless, the recoveries remained substantial. The fact that larvae migrating in swine or poultry organs and tissues have high infectivity in pigs even after preservation at 4 degrees C for 1 week, suggests that human infection with T. canis might easily occur following consumption of raw or undercooked dishes, either fresh or refrigerated, prepared from swine or poultry organs and tissues harbouring T. canis larvae. PMID- 15110410 TI - Evaluation of a larval development assay (DrenchRite) for the detection of anthelmintic resistance in cyathostomin nematodes of horses. AB - A larval development assay (LDA, DrenchRite) was evaluated to determine the effectiveness of this method in detecting anthelmintic resistance in cyathostomin nematodes of horses. A total of 15 horse farms from Georgia and South Carolina (USA) and Population S ponies from the University of Kentucky (USA) were included in this study. Nematode eggs were extracted from pooled fecal samples and placed into the wells of a DrenchRite plate for testing against thiabendazole (TBZ), levamisole (LEV) and 2 ivermectin (IVM) analogs (IVM-1, IVM-2). After a 7-day incubation larvae in each well were counted and data were analyzed by logistic regression. Resistance status of each farm for different drugs was determined in a separate study using a fecal egg count reduction test. LDA were performed on the 15 farms once, however, the Population S cyathostomins were assayed on 3 separate occasions to estimate the consistency of results between assays. Mean TBZ LC50 for oxibendazole resistant, suspected resistant and sensitive farms were 0.2015, 0.1625, and 0.1355 microM, respectively. For LEV, mean LC50 for PYR resistant, suspected resistant and sensitive farms were 1.590, 1.8018 and 1.4219 microM, respectively. All 15 farms had worms susceptible to IVM; mean LC50 for IVM-1 and for IVM-2 were 7.5727 and 87.9718 nM, respectively. A linear mixed model was fitted to the data to determine the relationship between LC50 and LC95 and resistance status for each farm. No meaningful relations were found. Consistency of assays varied between drugs, being best for TBZ and worst for LEV and IVM-1. All farms in this study had benzimidazole-resistant nematodes; therefore usefulness of DrenchRite for discriminating susceptibility versus resistance to this drug class could not be accurately assessed. Moreover, since all farms tested were sensitive to IVM and resistance to this drug class has not yet been reported in cyathostomins, it is not possible to assess accurately the usefulness of DrenchRite LDA for detecting IVM resistance at this time. Assay results for LEV suggest that LEV in a LDA does not yield data that is useful in estimating PYR efficacy in vivo. Based on results for PYR/LEV, the current high prevalence of benzimidazole resistance, no known cases of IVM resistance, and the sometimes extreme variation in results seen in many of the assays, DrenchRite LDA cannot be considered a useful tool for the diagnosis of resistance in cyathostomins of horses at present. PMID- 15110411 TI - Serologic response of red-legged partridges (Alectoris rufa) after oral inoculation with Toxoplasma gondii oocysts. AB - Thirty 5-month-old red-legged partridges (Alectoris rufa) reared in battery were divided into five groups: 4 birds in group A, 14 birds in group B, 4 birds in group C, 4 birds in group D and 4 birds in group E, and were inoculated orally with 10, 50, 10(2), 10(3) and 10(4) oocysts of the OV-51/95 strain of Toxoplasma gondii, respectively. During the experiment, blood samples from all birds were drawn every 3-7 days and at necropsy. Serologic response was measured by the modified agglutination test (MAT) and the latex agglutination test (LAT). One bird from each group was killed at 44, 58, 65 and 72 days after inoculation (DAI). From 72 DAI to the end of the experiment, surviving partridges from group B were killed at weekly intervals. The last partridges were sacrified 100 DAI. MAT was the most sensitive and specific test for detecting T. gondii antibodies in the birds. First positive titers were detected by MAT in all sera on 7 DAI, but titers by LAT did not appear until 13 DAI. Antibody titers detected by MAT on 7 DAI were higher in the partridges with the largest inocula (10(3) or 10(4) oocysts) than those inoculated with 10, 50 or 10(2) oocysts. All surviving birds developed a serologic response to T. gondii, with maximum titers of 512-32,768 in the MAT on 13-17 DAI, and positive titers persisted at least until 100 DAI. To the contrary, LAT reveals only very low antibody titers even in partridges inoculated with the highest dose of T. gondii. PMID- 15110412 TI - Transmission dynamics of the Echinococcus granulosus sheep-dog strain (G1 genotype) in camels in Tunisia. AB - Cystic echinococcosis, caused by Echinococcus granulosus, is highly endemic in North Africa and the Middle East. This paper examines the abundance and prevalence of infection of E. granulosus in camels in Tunisia. No cysts were found in 103 camels from Kebili, whilst 19 of 188 camels from Benguerden (10.1%) were infected. Of the cysts found 95% were considered fertile with the presence of protoscolices and 80% of protoscolices were considered viable by their ability to exclude aqueous eosin. Molecular techniques were used on cyst material from camels and this demonstrated that the study animals were infected with the G1 sheep strain of E. granulosus. Observed data were fitted to a mathematical model by maximum likelihood techniques to define the parameters and their confidence limits and the negative binomial distribution was used to define the error variance in the observed data. The infection pressure to camels was somewhat lower in comparison to sheep reported in an earlier study. However, because camels are much longer-lived animals, the results of the model fit suggested that older camels have a relatively high prevalence rate, reaching a most likely value of 32% at age 15 years. This could represent an important source of transmission to dogs and hence indirectly to man of this zonotic strain. In common with similar studies on other species, there was no evidence of parasite-induced immunity in camels. PMID- 15110413 TI - Effective treatment and control of biting lice, Felicola subrostratus (Nitzsch in Burmeister, 1838), on cats using fipronil formulations. AB - The efficacy of 0.25% fipronil spray (Frontline Spray, Merial), 10% fipronil spot on (Frontline Spot-on for Cats, Merial) and 10% fipronil/12% (S)-methoprene (Frontline Plus for Cats, Merial) against the biting louse Felicola subrostratus on cats was assessed and confirmed under laboratory conditions. A field study evaluated the efficacy of a single topical application of Frontline Spray, and spot-on against the parasite on cats. In the laboratory studies, animals were allocated based on pre-treatment louse counts to the treatment groups: (1) untreated control and (2) 10% fipronil spot-on in the exploratory study or (1) untreated control, (2) 0.25% fipronil spray, at 6 ml/kg; (3) 10% fipronil spot-on as per label and (4) 10% fipronil/12% (S)-methoprene as per label in the confirmation study. Cats in treatment groups 2-4 were treated twice topically on Days 0 and 28. No live F. subrostratus were found on cats treated with fipronil formulations at any post-treatment examination. The difference from controls was significant (P < 0.01) for each product at each examination. Based on whole body counts at Day 42, the efficacy of each product was determined to be 100%. In the field study, cats were allocated in strict order of presentation. Cats were randomly allocated to one of the three treatment groups: (1) propoxur collar (Bolfo, Bayer); (2) 0.25% fipronil spray, at 6 ml/kg and (3) 10% fipronil spot-on as per label. Cats were treated once topically on Day 0. Louse counts of cats treated with fipronil formulations were not different than those of cats receiving the propoxur collar. The efficacy was determined to be > 98% on Day 2 and 100% on Days 28 and 42 in all treatment groups. The results of these studies demonstrate that fipronil in topical formulations is effective for treatment and control of biting lice (F. subrostratus) infestations on cats. PMID- 15110414 TI - Scanning electron microscopic study of third-instar warbles in yak in China. AB - Scanning electron microscopic observations were performed to compare the structures of third-instar of three species of warbles (Hypoderma bovis, H. lineatum and H. sinense) that infest yak in China. The heads of the warbles consisted of mouths and opercular sutures with spinal bands between them. We could not detect any morphological differences in the shape of the warble's heads among these three species. The spiracular plate of H. bovis was strongly concaved and had numerous spines at the spiracular opening rims. The H. lineatum had flat spiracular plates and no spines on the spiracular plates. In the case of H. sinense the spiracular plate was as flat as that of H. lineatum but had small sized and fewer spines on the spiracular plates. At the ventral side on the tenth segment of the warble body of H. bovis the spinal band was absent. Although H. lineatum had only one band at the posterior border on the tenth ventral segment, H. sinense had two bands of spines at the anterior and posterior border on the tenth segment. PMID- 15110415 TI - Integrating genetics as practices of primary care. AB - This study examines the responses of general practitioners (GPs) in Vic., Australia to an increased emphasis on genetics in primary care. A qualitative analysis of focus group interviews with GPs in regional and metropolitan areas and one focus group interview with genetics experts showed that despite the emphasis placed on genetics by the experts, GPs remained ambivalent to the routine integration of genetics into general practice. This response from GPs has been noted in several studies and is most commonly attributed to GPs' lack of knowledge about genetics. In this study we argue that a 'cognitive deficit' understanding of the problem excludes many of the factors that GPs regard as important in relating genetics to primary care. We show that GPs' ambivalence emerges from how they situate genetics within practices of patient care and in relation to what they regard as good patient management. We found that GPs respond most enthusiastically to genetics and genetic testing if they feel it changes their management in ways they consider of benefit to their patients. GPs have specialist skills in managing the heterogeneity of patient care and these skills enable them to situate genetics relative to the overall needs of patients. Preparing GPs to 'do' genetics when the need arises by finding ways to make genetics information available to them as the need arises would facilitate the integration of genetics as practices of primary care. PMID- 15110416 TI - Optimising clinical practice in cancer genetics with cultural competence: lessons to be learned from ethnographic research with Chinese-Australians. AB - Hereditary cancer is about families, and clinicians and genetic counsellors need to understand the cultural beliefs of patients and families about cancer and inheritance. In the light of their kinship patterns Chinese-Australians were chosen for the present study, which aims to determine the explanatory models of inheritance, cancer, and inherited cancer, with a view to identifying the relationship between these culture-specific lay attributions and help-seeking behaviour, and to identify possible barriers to genetic counselling and testing. Qualitative ethnographically informed methodology involving semi-structured interview was used as a method to uncover latent beliefs held by the families who are represented by the subjects. In-depth interviews were conducted with 16 informants of Chinese ethnicity, who had been recruited through two major Sydney familial cancer clinics. We report the attributions of cancer in general, then on inheritance, kinship, genes and genetics and then focus on the way in which these beliefs come together around hereditary cancer. The majority of informants, despite high acculturation and belief in biomedical explanations about hereditary cancer, also acknowledged the influence of traditional family Chinese beliefs, where 'inheritance' and 'genetics' were related to retribution for ancestral misdeeds and offending ancestors. Extensive mismatch of attributes and beliefs were identified in those who attended the clinic and senior family members, creating barriers to optimal service utilisation. Three traditional patterns of beliefs were identified: (a) father and mother contributed in equal share to one's genetic makeup, linked to the ying-yang theory; (b) the dominance of life force (yang chi) and the shaping of genes were transmitted through the paternal line; and (c) natural and supernatural forces operated in the cause of hereditary cancer. The study provided guidance for clinical practice. Exploration and acknowledgement of family beliefs, regardless of cultural background and therefore avoiding stereotyping, can enable the clinician to work with the whole family-those who hold Western attributions, those who maintain traditional notions of genetics and inheritance, and those who incorporate both into their belief systems-and provide effective clinical services. Further ethnographic studies are needed, focusing on the Chinese groups who do not attend the clinic and those with lower acculturation and educational levels. PMID- 15110417 TI - Socioeconomic differences in cancer screening participation: comparing cognitive and psychosocial explanations. AB - This paper compares psychosocial and cognitive models of socioeconomic variation in participation in screening for colorectal cancer. The psychosocial model suggests that factors such as higher stress and lower social support explain, in part, why people from lower socioeconomic status (SES) environments are less likely to participate in screening. The cognitive model suggests that beliefs about cancer risk and screening will play an important part in differential participation. In practice both sets of factors may contribute to explaining socioeconomic differentials. The data for these analyses are drawn from a randomised controlled trial of colorectal cancer screening (the UK Flexible Sigmoidoscopy Trial). The participants are from the Scottish centre, where recruitment was stratified to generate a socioeconomically diverse sample. The dependent variable was interest in attending screening. A questionnaire covering demographic status, psychosocial and cognitive factors as well as interest in screening was sent to 10,650 adults. The results showed the predicted SES gradient in interest. There were also SES differences in both psychosocial and cognitive variables. A series of logistic regression models were used to test potential mediators of the association between SES and interest in attending screening by successively including psychosocial factors, cognitive factors, and then both, in the equation. Only the inclusion of the cognitive variables significantly reduced the variation associated with SES, providing better support for the cognitive than the psychosocial model. PMID- 15110418 TI - Women, family demands and health: the importance of employment status and socio economic position. AB - Although it is generally assumed that women engaged in paid work have better health than full-time homemakers, little is known about the situation in Southern European countries like Spain or about differences in the impact of family demands by employment status or the potential interaction with educational level. The objectives of this study are to analyse whether inequalities in health exist among housewives and employed women, and to assess whether the relationship between family demands and health differs by employment status. Additionally, for both objectives we examine the potential different patterns by educational level. The data have been taken from the 1994 Catalonian Health Survey (Spain). The sample was drawn from all women aged 25-64 years who were employed or full-time homemakers and married or cohabiting. Four health indicators (self-perceived health status, limiting long-standing illness, chronic conditions and mental health) and two health related behaviours (hours of sleeping and leisure-time physical activity) were analysed. Family demands were measured through household size, living with children under 15 and living with elderly. Overall, female workers had a better health status than housewives, although this pattern was more consistent for women of low educational level. Conversely, the health related behaviours analysed were less favourable for workers, mainly for those of low educational level. Among workers of low educational level, family demands showed a negative effect in most health indicators and health related behaviours, but had little or no negative association at all in workers of high educational level or in full-time homemakers. Moreover, among women of low educational level, both workers and housewives, living with elderly had showed a negative association with poor health status and health related behaviours. These results emphasise the need of considering the interaction between family demands, employment status and educational level in analysing the impact of family demands on women's health as well as in designing family policies and programmes of women's health promotion. PMID- 15110419 TI - Understanding the role of mediating risk factors and proxy effects in the association between socio-economic status and untreated hypertension. AB - The association between socio-economic status (SES) and untreated hypertension varies according to a country's level of development and racial/ethnic group. We sought to confirm this variation in women from China and the United States (US) as well as to investigate the impact of SES on several mediating risk factors. We also investigate the extent to which SES explains racial/ethnic differences in untreated hypertension in the US. We used cross-sectional data from 1814 non pregnant women in China (China Health and Nutrition Survey (CHNS), 1997) and 3266 non-pregnant women in the United States (National Health and Nutrition Examination Survey (NHANES III), 1988-1994) respectively. A variety of statistical modelling techniques was used to predict untreated hypertension as a function of several mediating factors and to simulate the impact of changes in SES. The age-adjusted prevalence of untreated hypertension was significantly higher (p<0.01) for low-income White and Black women compared to Mexican American or Chinese women. Untreated hypertension was not significantly associated with income or education in Mexican Americans or women in China. Obesity and light physical activity had the largest mediating effect on the association between SES and untreated hypertension for all racial/ethnic groups. However, this effect was not as strong as the proxy effect of income and education. SES did not completely explain racial/ethnic differences in hypertension in the US. While SES was more strongly associated with hypertension in Blacks than Whites, Blacks were still 1.97 (95% CI 1.47-2.64) times more likely to have untreated hypertension than Whites after adjusting for SES differences. The association between SES and untreated hypertension varied by country and racial/ethnic group. An important explanation for this variation was the differential effect of SES on mediating risk factors. SES disparities between Whites and Blacks in the US partly explain differences in the prevalence of untreated hypertension between these racial/ethnic groups. PMID- 15110420 TI - AIDS-talk in everyday life: the presence of HIV/AIDS in men's informal conversation in Southern Malawi. AB - Malawi is one of the world's most AIDS-afflicted countries. In order to cope with the AIDS pandemic, we must know what the people most at risk think about it, how they evaluate their situation and that of their community, and what actions they would consider adopting to lower their risk. However, the main research methods in studying attitudes-surveys and questionnaires-have only a limited ability to capture what people think about AIDS. In order to get a more naturalistic perspective on attitudes towards AIDS from 1999 to 2001 six Malawian research assistants who lived in rural villages were asked to keep journals in which they wrote down information about all the conversations they participated in or witnessed in which the topic of AIDS surfaced in any way. The conversations ranged from graveside condolences following a funeral to stories told during men only beer-drinking sessions, to women chatting on the bus. In this paper, I analyse these journals in order to see how men talk about AIDS in naturalistic settings, what they perceive as the impact of the AIDS epidemic, and how they understand AIDS risk. PMID- 15110421 TI - Secretive females or swaggering males? An assessment of the quality of sexual partnership reporting in rural Tanzania. AB - In population-based surveys on sexual behaviour, men consistently report higher numbers of sexual partners than women, which may be associated with male exaggeration or female under-reporting or with issues related to sampling, such as exclusion of female sex workers. This paper presents an analysis of data collected in the context of a longitudinal study in rural Tanzania, where a sexual partnership module was applied to all participating men and women in the study population. Since the study design included all men and women of reproductive ages and did not involve sampling, these data provide a unique opportunity to compare the consistency of aggregate measures of sexual behaviour between men and women living in the same villages. The analysis shows that non marital partnerships were common amongst single people of both sexes--around 70% of unmarried men and women report at least one sexual partner in the last year. However, 40% of married men also report having non-marital partners, but only 3% of married women did so. Single women reported about half as many multiple partnerships in the last year as men. Under-reporting of non-marital partnerships was much more common among single women than among married women and men. Furthermore, women were more likely to report longer duration partnerships and partnership with urban men or more educated men than with others. If a woman reports multiple partners, biological data indicate that she is at high risk of contracting HIV. For men, however, there is only a weak association between number of partnerships and the risk of HIV, and it cannot be excluded that men, especially single men, exaggerate the number of sexual partners. PMID- 15110422 TI - Violent deaths among women of reproductive age in rural Bangladesh. AB - The objectives of this paper are to investigate levels and trends in mortality due to violence in women of reproductive age and the social and demographic factors associated with such mortality. The study took place in Matlab, a rural sub-district in Bangladesh between 1982 and 1998. The data were furnished by a longitudinal population-based demographic surveillance system located in that area. A case-control design study was used out to identify factors associated with death due to violence, and data from death registration forms were analyzed. The death rate due to violence, defined in terms of suicides and homicides, was higher among women than men. Death rates from violence remained at the same level during the study period while death rates from other causes decreased. Young, not yet married women were a high risk group with respect to death from violence. Oppression, physical and mental abuse by husbands and relatives often preceded suicides and homicides. The disadvantaged position of women in Bangladesh society is the key underlying social cause of the violence that occurred. In order for violence against women to decrease, improvement in the social position of women is essential. PMID- 15110423 TI - Dependency, democracy, and infant mortality: a quantitative, cross-national analysis of less developed countries. AB - This study presents quantitative, sociological models designed to account for cross-national variation in infant mortality rates. We consider variables linked to four different theoretical perspectives: the economic modernization, social modernization, political modernization, and dependency perspectives. The study is based on a panel regression analysis of a sample of 59 developing countries. Our preliminary analysis based on additive models replicates prior studies to the extent that we find that indicators linked to economic and social modernization have beneficial effects on infant mortality. We also find support for hypotheses derived from the dependency perspective suggesting that multinational corporate penetration fosters higher levels of infant mortality. Subsequent analysis incorporating interaction effects suggest that the level of political democracy conditions the effects of dependency relationships based upon exports, investments from multinational corporations, and international lending institutions. Transnational economic linkages associated with exports, multinational corporations, and international lending institutions adversely affect infant mortality more strongly at lower levels of democracy than at higher levels of democracy: intranational, political factors interact with the international, economic forces to affect infant mortality. We conclude with some brief policy recommendations and suggestions for the direction of future research. PMID- 15110424 TI - Wilp Wa'ums: colonial encounter, decolonization and medical care among the Nisga'a. AB - The Nisga'a Nation of Northwestern British Columbia have been pioneers in the area of obtaining administrative control over health services. This would seem to mark the end of medical colonialism for this First Nation. But the author argues that health program devolution, in this case, was part of a longer tradition of incorporating aspects of non-Native medicine in a way that supported Nisga'a social structure. Nevertheless, the author argues that health program devolution is part of the process of decolonization since it has supported the traditional social structure, enhanced community self-esteem and provided an opportunity for the locus of control to shift from the medical profession and the federal government to Nisga'a people. This paper sets the development of Nisga'a-centered health care in a historical context that sees the Nisga'a exerting a resistive will in the context of medical colonialism in twentieth century British Columbia. PMID- 15110425 TI - Use of a visual analogue scale in a daily patient diary: modelling cross sectional time-series data on health-related quality of life. AB - An economic evaluation of beta interferon therapy collected quality of life data from 62 people with relapsing-remitting multiple sclerosis, currently in remission. Each completed a postal questionnaire consisting of the Euroqol EQ-5D questionnaire, the MSQOL-54 (a disease-specific measure incorporating the SF36) and other data including health services use. This was completed at the beginning and end of 6-weeks during which a daily diary was kept. The diary asked about daily activities, symptoms experienced, the impact of symptoms, general health status and current health status using the Euroqol Visual Analogue Scale (EQ VAS). The diary had an excellent completion rate, producing data with good face validity. The resulting cross-sectional time-series data provide information about the stability and variability of the EQ VAS in repeated measurement and its sensitivity to health state changes. EQ VAS scores were stable at the population level over time; greater variation in scores was observed between rather than within individuals. Panel data techniques are used to relate EQ VAS scores to recorded symptoms and baseline general health status, paying particular attention to the role of individual heterogeneity and the dynamic nature of responses. The EQ VAS was sensitive to the presence of symptoms, their severity and their type. It is concluded that appropriately analysed panel data can provide insights useful in the measurement of health-related quality of life. PMID- 15110426 TI - Geographical variation in acute psychiatric admissions within New York City 1990 2000: growing inequalities in service use? AB - The paper analyses geographical variations in use of acute psychiatric inpatient services within New York City and how these have changed from 1990 to 2000. We review literature suggesting reasons for the variations observed. Data from the New York State Department of Health Statewide Planning Research and Cooperative System were combined with population census data to produce age standardized ratio indicators of admissions and of bed days, as measures of use of general hospitals for psychiatric conditions, by males aged 15-64, in Zip Code Areas of New York City, in 1990 and 2000. Geographical variations in hospital use were related to proximity to general hospitals with psychiatric beds and to socio economic status of local populations (as recorded in the 1990 and 2000 population censuses). Areas close to psychiatric hospitals areas show high admission levels. Controlling for this, Zip Code Areas with higher concentrations of poverty, of African American residents or of persons living alone were associated with relatively high admission ratios. These relationships vary somewhat between diagnostic groups. Area inequalities in standardized admission ratios persisted and widened between 1990 and 2000, and the highest hospital admission ratios were increasingly concentrated where social and economic disadvantage was greatest. Various possible reasons for this trend are explored. We conclude that increasing intensity of poverty in disadvantaged areas is not likely to provide an explanation and that the trends are more likely to result from changes in hospital management and funding affecting access to hospital services. PMID- 15110427 TI - The health-promoting family: a conceptual framework for future research. AB - There has so far only been little research attention given to how families actively engage in promoting their health in everyday life. In this paper a theoretical framework is proposed for studies of the 'health-promoting family' with particular focus on children's health and well-being. This paper sets out a conceptual model for understanding how the family can play a part in promoting both the health of children and children's capacities as health-promoting actors. It draws on contemporary social science approaches to health, the family and children, suggesting a new emphasis on the family's ecocultural pathway, family practices and the child as a health-promoting actor. PMID- 15110428 TI - The Israeli kibbutz as a venue for reduced disability in old age: lessons from the Cross-sectional and Longitudinal Aging Study (CALAS). AB - It is important to identify ways to moderate disability in old age. We assessed whether the kibbutz way of life results in reduced disability by examining risk factors for disability in three comparable populations: kibbutz members (lifetime kibbutz exposure); parents of kibbutz members who came to live on the kibbutz in old age due to health and social needs (old age exposure); and comparable Israelis in the general population (no exposure). Kibbutz members were less disabled, defined as needing help with at least one of five activities of daily living, than the other groups. Kibbutz members had 30% lower risk of disability, after controlling for sociodemographics and social networks, health and health behaviors, and life history and background. Introducing length of residence in current home and death of a child to the analysis reduced this finding to non significance. Overall, risk of disability was significantly higher for older age groups, women, homemakers, people with more comorbid conditions, those with more children, and those with a child that had died, while risk of disability was significantly lower for those with larger social networks, those who had engaged in physical activity at midlife, and those who had lived in their homes longer. Since occupations on the kibbutz (primarily agricultural and blue collar) have equal pay, and kibbutz members have complete economic security, our results suggest that it is not such occupations themselves but their association with low incomes that have contributed to previous associations of blue collar and agricultural occupations with poor health and high disability. Lower disability among kibbutz members may be due to the social, economic, and instrumental support provided on the kibbutz, as well as to an active life style, suggesting features of kibbutz life that can be replicated elsewhere to reduce disability. PMID- 15110429 TI - Traffic stress, vehicular burden and well-being: a multilevel analysis. AB - This study examined whether health is associated with individually perceived traffic stress and as well as ecologically measured vehicular burden using multi level analysis. Data from the Chinese American psychiatric epidemiologic study (N = 1503) are linked to data from the 1990 Census in the United States. Hierarchical linear modeling was used to analyze the cross-sectional relationship between traffic stress, neighborhood conditions, depression and health status. Perceived traffic stress is associated with both general health status and depression in multivariate multilevel models, such that persons reporting traffic stress had lower health status and more depressive symptoms. Further, there is an interaction between vehicular burden and traffic stress for both health outcomes. Persons who lived in areas with greater vehicular burden and who reported the most traffic stress also had the lowest health status and greatest depressive symptoms. These findings suggest that traffic stress may represent an important factor that influences the well-being of urban populations, and that studies which examine factors at only one level (either individual level only or ecological level only) may underestimate the effect of the social environment. PMID- 15110430 TI - Popular perceptions of tobacco products and patterns of use among male college students in India. AB - This paper examines popular perceptions of tobacco products and describes patterns of use among college youth in Karnataka, India. Data are drawn from 25 key informant interviews and six focus groups with male and female college students, interviews with shopkeepers, observational data on youth tobacco consumption, and a college-based survey. The survey was administered to 1587 males attending eleven colleges. Forty-five percent (n = 716) of college students surveyed had used tobacco products. Thirty-six percent (n = 573) had tried cigarettes, 10% (n = 157) had tried bidis, and 18% (n = 290) had tried gutkha. Tobacco consumption among smokers was low; for daily smokers, the mean number of cigarettes smoked was 6 per day. Students attending professional colleges, including engineering, medicine, and law were significantly more likely to have ever smoked and to be daily smokers when compared to students enrolled in other courses of study. In interviews, male students noted that smoking a cigarette enhanced one's manliness, relieved boredom, and eased tension. Although female students interviewed were non-smokers, several suggested that in the future, smoking might be an acceptable behavior among college-going females. When asked about their perceptions of smoking among youth in Western countries, the majority of students believed that three-quarters of male and female youth in the West smoked. This perception has been largely formed through media images, including satellite television and films. With regard to addiction, it was widely believed that filter-tipped cigarettes were one of the most addictive products because they are made of better quality tobacco, and are milder and smoother to smoke. Therefore, a person could easily smoke more of them, which would lead to addiction. Another widely held belief was that the more expensive the cigarette, the less harmful it was for one's health. PMID- 15110431 TI - Use of benzodiazepine drugs and perceived job stress in a cohort of working men and women in Belgium. Results from the BELSTRESS-study. AB - The aim of the Belstress Study was to see whether use of benzodiazepines is associated with perceived job stress as measured by Karasek's job-strain model. This model has as its central tenet that the most adverse health outcomes are to be expected in high strain jobs characterized by high job demands and low job control. An extension of the model states that the most noxious combination is high job demands, low control and low social support at work. Sample subjects were recruited from 25 Belgian companies between 1994 and 1998, and cover a wide range of occupations. A 5.6% of 16,094 men and 9.3% of 5012 women aged 35-59 years report use of benzodiazepines during the last month. A clear association is displayed between self-reported use of benzodiazepines and a high strain job compared to a low strain job (men: OR=1.93, 99% CI=1.4-2.6; women: OR=1.66, 99% CI=1.0-2.7), after adjustment is made for socio-demographic confounders (age, level of education, occupational group, employment sector, living situation). The independent association with quartile level of job demands is a striking feature (men: OR of highest quartile compared to lowest quartile group=1.91, 99% CI=1.4 2.6; women: OR=1.99, 99% CI=1.3-3.1). In men, an inverse association with quartile level of job control is observed (OR= 0.65, 99% CI=0.5-0.9) whereas in women a clear tendency in that direction is displayed (OR=0.62, 99% CI=0.4-1.1). The association with low social support is less clear; an independent association between use of benzodiazepines and iso-strain was observed particularly in men. PMID- 15110432 TI - Safeguarding children and the future of the nursing and midwifery council. PMID- 15110433 TI - The impact of clinical placement location on nursing students' competence and preparedness for practice. AB - The challenges confronting nurses in today's health care environments have highlighted the necessity for graduating students to feel both competent and prepared for practice. The aim of the aspect of the study reported in this article was to determine the relationship between the location of clinical placements and competence and preparedness for practice from the perspective of the nursing students. A quasi-experimental design using a pre-test post-test survey was used. The population consisted of all final year Bachelor of Nursing students at Queensland University of Technology, who undertook their clinical placement in either a rural or metropolitan location. The response rate was 65% in the pretest (n = 137) and 57% in the posttest (n =121). The results of the study highlight the importance of both rural and metropolitan clinical environments in developing not only student competency but also students' satisfaction with their clinical experience. The results suggest that the selection of a rural placement is more likely to occur when students feel competent, confident and organised about their clinical experience. Strategies to increase students' perceptions of competence and confidence to undertake a rural placement are likely to increase students' selection to 'go rural'. PMID- 15110434 TI - The educational preparation of nursing staff undertaking pre-assessment of surgical patients--a discussion of the issues. AB - This article discusses the education and training needs of nursing staff undertaking the pre-assessment of surgical patients in the context of role development within health care. The authors undertook this piece of work as part of a project to inform the development of a competency framework for use by registered nurses working in the field of pre-assessment screening and preparation of surgical patients. A selective review of the literature on the education and training needs of pre-assessment nurses, and nurses undertaking expanded roles has been undertaken and some of the emerging issues are debated. The nature of current preparation for these roles and the apparent lack of any consistent approach to education and training for staff adjusting their scope of professional practice is debated. A brief review of the nature of competence and some of the different models including occupational competence models are presented. Finally, the implications of these for the development of a strategy for the acquisition and demonstration of knowledge and skills in pre-assessment nursing staff are presented. PMID- 15110435 TI - How the metaphor of a gap between theory and practice has influenced nursing education. AB - The metaphor of a gap between theory and practice is firmly established in nursing education. Furthermore, it is embedded in conventional professional wisdom to the extent that when it used in professional reports, academic writing or in the conversations amongst nurses, it often requires no further explanation. The notion of a gap between theory and practice has proven to be a most fertile and an especially helpful concept which has enabled educators, clinicians and students to devise practical strategies, intended to resolve problems associated with the gap. However, that nursing has so enthusiastically embraced the notion of a gap between theory and practice may also have inhibited creativity in the design and delivery of programmes and thereby missing other opportunities for student nurses to learn from practical contexts. PMID- 15110436 TI - Distance learning in post-qualifying nurse education. AB - There has been a prolific development of distance education in nursing during the previous decade. This has not been supported by an increase in research activity to generate the knowledge that is required to support these developments. In response to this, within a wider action research approach, a team of academics undertook a study to examine post-qualifying nurses' experiences of distance learning, and the impact of distance education on lecturing staff. Five teaching modules, delivered through distance learning, were selected for the study using a matrix sampling technique. Data collection took place at key points during these modules: at the commencement of study students and lecturers were asked to maintain a diary account of their experiences, students were sent questionnaires at the mid point of the module ( n=115, 30.4% return rate), and at the end of the module the students and lecturers participated in one of nine group interviews (students =15; LECTURERS = 15). Findings suggest that distance education is an effective learning medium that enables students to balance learning with personal and work commitments. Students and lecturers face a transitional period when embarking on distance education and this involves the development of new understandings and skills to successfully adapt to this approach to education. PMID- 15110437 TI - Self direction in a problem based learning program. AB - Baccalaureate nursing graduates must enter their professional careers with the understanding and ability to engage in continuing learning in order to ensure competence in professional practice. The graduate's ability to be self directed in learning is an important indication of the ability to pursue continuing learning as a practitioner. It has been surmised that Problem Based Learning nursing programs facilitate the development of abilities to become self directed in learning. A quasi-experimental design using the self directed learning readiness scale (SDLRS) was used to examine the self directed learning (SDL) of students at the beginning and end of one year in a problem based learning (PBL) program. In order to complement the quantitative data, year end focus group interviews were held with students to explore their experiences with SDL. Students demonstrated average self directed learning readiness scores at the beginning and end of the first year in the PBL program. Although students did not demonstrate any increase in SDLRS at the end of one year in a PBL program, they were able to describe themselves as having developed many of the documented characteristics associated with self directed learners. PMID- 15110438 TI - Study of critical thinking skills in nursing students and nurses in Japan. AB - The purpose of this study was to measure the dimensions of critical thinking (CT) of nursing students at baccalaureate nursing program and registered nurses at general hospital in Japan. Relevant literature on the current environment of Japanese nursing practice and education is reviewed as it provides the background to the key aspects of dimensions of Japanese nurses' and students' CT. The California Critical Thinking Disposition Inventory (CCTDI) was used to measure the dimensions of CT skills. The convenience sample consisted of three small groups: generic students (n=82) including freshmen and juniors; transfer students (n=16) at selected baccalaureate nursing program; and registered nurses (n=67) at selected general hospital were administered CCTDI. Descriptive statistic indicated that all groups had an ambivalent disposition towards CT in majority of sub-scale while they scored a positive disposition towards CT on several sub scales. A one-way ANOVA indicated that registered nurses scored lower than other two groups of baccalaureate students on the total score and several sub-scale score. The outcomes of this study propose recommendations regarding curriculum review for Japanese nursing education and reflection on professional boundaries for Japanese nursing practice. PMID- 15110439 TI - Biological and physiological knowledge and skills of graduating Finnish nursing students to practice in intensive care. AB - This study describes the basic biological and physiological knowledge and skills of graduating nurse students in Finland against the requirement of their being able to practice safely and effectively in intensive care. The study describes also their interest and willingness to work in intensive care. Measurements were based on the Basic Knowledge Assessment Tool (BKAT-5) developed by Toth in the United States. The sample consisted of 130 nursing students graduating in December 2001 and January 2002. The data were analysed statistically. The students were most knowledgeable in the areas of appropriate precautions, living will and medical calculation, followed by neurology and endocrinology. Scores were poorest for pulmonary, gastrointestinal and cardiovascular knowledge. Intensive care studies and the desire to work in intensive care correlated significantly with the respondents' basic intensive care knowledge. It is important for nursing education to concentrate on developing those areas of intensive care studies where the performance of students is weakest. PMID- 15110440 TI - Can nursing facility staff with minimal education be successfully trained with computer-based training? AB - This study compares the effects of computer-based and instructor-led training on long-term care staff with a high school education or less on fire safety knowledge, attitudes, and practices. Findings show that both methods of instruction were effective in increasing staff tests scores from pre- to posttest. Scores of both groups were lower at follow-up three months later but continued to be higher than at pretest. Staff with a high school education increased scores more than those without a high school diploma. PMID- 15110441 TI - Evaluating the service quality of undergraduate nursing education in Taiwan- using quality function deployment. AB - This study applies quality function deployment (QFD) techniques to evaluate the quality of service of undergraduate nursing education in Taiwan from the perspective of nursing students. Survey data from 560 undergraduate nursing students at four Taiwanese universities were subjected to QFD analysis in order to identify the quality characteristics most highly valued by students, the elements of educational service they consider most important and least important, and relationships/discrepancies between student quality requirements and institutional service elements. Results show that students value traditional elements of nursing education - clinical practice and lectures - more highly than recent additions such as computer-aided instruction and multimedia teaching. Results also show that students are looking for quality primarily in the area of faculty characteristics. The implication is that institutions which provide nursing education should not neglect the importance of investing in faculty when they are seeking to upgrade the quality of their programs. Further QFD studies are recommended to evaluate the quality of nursing education from the perspective of preceptors and nurses who help to train students in clinical settings. PMID- 15110442 TI - Flexible learning for postgraduate nurses: a basis for planning. AB - This paper describes a survey undertaken with postgraduate nursing students in a New Zealand University based School of Nursing in 2002 to establish their access to and use of computers and information technology for study. Whilst there is minimal flexibility and use of technology to support student learning for postgraduate nurses in the University's School of Nursing presently, the University proposes increasing flexibility across all courses. This is in part a response to the increased internationalisation of education and developments in technology affecting programme design, delivery and support that can benefit teachers and students. The findings of this survey form a basis for planning the introduction of flexible learning. Results indicate that not all students have convenient access to technology for study purposes, nor are they at the same level in terms of using technology. The implications of the survey for planning flexible learning, including the need to be explicit about requirements for access to technology and upskilling opportunities are discussed. PMID- 15110443 TI - Thinking. AB - This paper argues that thinking is assumed within nursing education. There are strategies to promote thinking: reflective practice, critical analysis and problem solving. I suggest that by categorising thinking into such boxes there may be a danger of limiting the rich possibilities of simply 'thinking'. The writings of Heidegger (1889-1976) are cited, highlighting the need to 'call' thinking, and to meditate or ponder on the things that matter. The paper comes from research that asked the question 'What calls for thinking in postgraduate education?' Findings reveal examples of teachers and students recalling 'thinking experiences' but also suggest there is a danger that students do not have time to think in a busy classroom situation. It appears that thinking is more likely to happen outside of the classroom, with peers, in assignment writing, or when thoughts simply come. The challenge to nursing education is that not only may teachers be limiting students thinking opportunities, but they may be directing thinking in a way that maintains the status quo. If nursing is to equip itself as a dynamic profession and take initiative for shaping its own future, then close attention must be paid to enabling thinking. PMID- 15110445 TI - Prediction of radiation pneumonitis by dose - volume histogram parameters in lung cancer--a systematic review. AB - BACKGROUND AND PURPOSE: To perform a systematic review of the predictive ability of various dose-volume histogram (DVH) parameters (V(dose), mean lung dose (MLD), and normal tissue complication probability (NTCP)) in the incidence of radiation pneumonitis (RP) caused by external-beam radiation therapy. METHODS AND MATERIALS: Studies assessing the relationship between CT-based DVH reduction parameters and RP rate in radically treated lung cancer were eligible for the review. Synonyms for RP, lung cancer, DVH and its associated parameters (NTCP, V(20), V(30), MLD) were combined in a search strategy involving electronic databases, secondary reference searching, and consultation with experts. Individual or group data were abstracted from the various reports to calculate operating characteristics and odds ratios for the different DVH metrics. RESULTS: A total of 12 published studies and two abstracts were identified. Eleven studies assessed V(dose), seven assessed MLD, and eight assessed NTCP. Nine studies exclusively analyzed the association between various DVH metrics and RP risk. Five studies also analyzed other patient, tumor, and treatment variables in conjunction with standard DVH metrics. A direct comparison between studies and the generation of summary statistics (i.e. meta-analysis) could not be achieved due to significant predictive and outcome variable heterogeneity. Most studies did show an association between DVH parameters and RP risk. However, overall accuracy, sensitivity, specificity, and positive predictive value were generally poor to fair for all three classes of DVH metrics. CONCLUSIONS: An association between DVH parameters and RP risk has been demonstrated in the literature. However, the ideal DVH metric with excellent operating characteristics, either alone or in a model with other predictive variables, for RP risk prediction has not yet been identified. Several recommendations for reporting and conduct of future research into the association between DVH metrics and RP risk are provided. PMID- 15110444 TI - Incidental irradiation of nodal regions at risk during limited-field radiotherapy (RT) in dose-escalation studies in nonsmall cell lung cancer (NSCLC). Enough to convert no-elective into elective nodal irradiation (ENI)? PMID- 15110446 TI - Literature-based recommendations for treatment planning and execution in high dose radiotherapy for lung cancer. AB - BACKGROUND AND PURPOSE: To review the literature on techniques used in high-dose radiotherapy of lung cancer in order to develop recommendations for clinical practice and for use in research protocols. PATIENTS AND METHODS: A literature search was performed for articles and abstracts that were considered both clinically relevant and practical to use. The relevant information was arbitrarily categorized under the following headings: patient positioning, CT scanning, incorporating tumour mobility, definition of target volumes, radiotherapy planning, treatment delivery, and scoring of response and toxicity. RESULTS: Recommendations were made for each of the above steps from the published literature. Although most of the recommended techniques have yet to be evaluated in multicenter clinical trials, their use in high-dose radiotherapy to the thorax appears to be rational on the basis of current evidence. CONCLUSIONS: Recommendations for the clinical implementation of high-dose conformal radiotherapy for lung tumours were identified in the literature. Procedures that are still considered to be investigational were also highlighted. PMID- 15110447 TI - Local control and recurrence of stage I non-small cell lung cancer after carbon ion radiotherapy. AB - BACKGROUND AND PURPOSE: For a proper evaluation of the relationship between carbon ion beam dose escalation and local control in the 81 patients with 82 lesions of stage I non-small cell lung cancer, we have identified the incidence of in-field recurrence by collating the dose distribution with the CT images. PATIENTS AND METHODS: Eighteen fractions over 6 weeks for 47 patients (48 lesions) and nine fractions over 3 weeks for 34 patients were applied in the carbon dose escalation method from 59.4 to 95.4 gray equivalents (GyE) by a 10% increment and from 68.4 to 79.2GyE by a 5% increment, respectively. The radiation target consisted of primary tumor. Image analysis of the patients with local recurrence was systematically performed after the treatment by focusing attention on the enhanced thin slice CT images of the primary lesion. By superimposing the dose distribution on the planning CT image and marking the anatomically identified loci of recurrence, it was possible to establish the relationship between the dose distribution and the incipient loci of recurrence and to classify the recurrence patterns from the differences in the recurrence loci. RESULTS: Local recurrence was found in 19 (23.2%) out of a total of 82 lesions. It is possible to distinguish between three recurrence patterns: Pattern 1 representing marginal recurrence and patterns 2a and 2b, which are both instances of in-field recurrence. In pattern 1, four recurrences take place from a region on the upper tumor margin. In pattern 2a, 13 recurrences take place from the center of the tumor. In pattern 2b, two recurrences take place from the near center of the tumor. CONCLUSIONS: For the 15 in-field recurrence lesions (patterns 2a and 2b) after excluding the four marginal recurrence lesions (patterns 1), we have established that the local control shows dose-dependence. Based on this, we have determined the optimal therapeutic dose. PMID- 15110448 TI - Escalated hyperfractionated accelerated radiation therapy for locally advanced non-small cell lung cancer: a clinical phase II trial. AB - BACKGROUND AND PURPOSE: To evaluate the feasibility, toxicity and the efficacy for locally advanced non-small cell lung cancer (NSCLC) treated with escalated hyperfractionated accelerated radiation therapy (EHART) combined with chemotherapy. PATIENTS AND METHODS: The EHART consisted of irradiation delivered twice per day with >6-h interval and five treatment days per week. In the first and second weeks, 1.2 Gy/fraction b.i.d, was given, and then 1. /fraction b.i.d in the third week; 1.4 Gy/fraction b.i.d in the fourth week; and 1. /fraction b.i.d in the fifth week, respectively. The total tumor dose delivered was 66 Gy/50 fractions/5 weeks. All patients received neoadjuvant and adjuvant chemotherapy. The chemotherapeutic regimen used was either MVP (mitomycin C, vindesine, cis-platinum), or EP (etoposide and cis-platinum). RESULTS: From February 1997 to February 1999, 73 eligible patients were registered. All were in stage IIIb with median age of 60 years (33-70). Of the 73 patients, 12 cases were withdrawn from the study due to Grade (Gr) III acute complications, distant metastases, or intercurrent diseases. Sixty-one patients completed the combined treatment as planned. A median of 4 cycles of chemotherapy (1-7) was administered and 66 Gy/50 fractions/36 days was delivered finally. The most common acute complication was radiation esophagitis, which occurred in 56 cases (77%), with Gr III in 11 cases (15%). Twenty-nine patients (40%) had acute pulmonary toxicity; with Gr III in 6 cases (8%). The median survival time was 13 months for the entire group. The 1-year and 2-year survival rates were 51 and 10%, respectively. Of the 61 patients who finished EHART, 34 were found to have locoregional progression. Thirty-two patients failed inside radiation fields, and 2 patients, outside radiation fields. The 1-year and 2-year locoregional progression-free rates were 71 and 34%, respectively. The 1-year and 2-year distant metastasis rates were 57 and 84%, respectively. CONCLUSIONS: EHART combined with chemotherapy could be tolerated by most of the stage IIIb NSCLC patients with acceptable complications. Locoregional control was improved, but the long survival was not prolonged significantly predominantly due to distant metastases. PMID- 15110449 TI - Escalated dose for non-small-cell lung cancer with accelerated hypofractionated three-dimensional conformal radiation therapy. AB - BACKGROUND AND PURPOSE: To prospectively assess the feasibility and efficacy of a hypofractionated accelerated radiotherapy regimen (72 Gy in 24 daily fractions, 3 Gy per fraction) in patients (pts) with non-resectable non-small-cell lung cancer (NSCLC). MATERIAL AND METHODS: We included 25 pts with a histologically or cytologically proven NSCLC, with KPS > or = 70 and < or =10% weight loss over prior three months, and with tumour stage I/II medically inoperable (9 pts) or non-resectable stage III a/b without pleural effusion (16 pts). Eleven pts received induction chemotherapy. No more than 30% of the combined lung volume could receive more than 25 Gy and the maximal biological effective dose to the spinal cord was maintained below 44 Gy. RESULTS: No grade-4 acute toxicity event was reported. Two pts had a treatment break because of grade-3 acute oesophagitis. Twenty-two pts were evaluable for long-term toxicity (median follow up=9.7 months, range 4 to 30.2 months). There were 4 Grade-1 pulmonary and 2 Grade-1 oesophageal long-term toxicity events. Twenty-two pts were evaluable for tumour response with 7 complete and 8 partial responses, 5 stable diseases and 2 progressive diseases. The actuarial 1-year overall and thoracic-progression-free survival rates were 68% and 72% respectively. CONCLUSIONS: This study demonstrates the feasibility of the experimental radiotherapy schedule, however more data are needed to confirm its efficacy. PMID- 15110450 TI - Radiation treatment of lung cancer--patterns of practice in Canada. AB - BACKGROUND AND PURPOSE: To assess the patterns of practice among Canadian radiation oncologists who treat lung cancers. PATIENTS AND METHODS: A questionnaire detailing different aspects of radiation treatment of lung cancer was mailed to all radiation oncologists treating lung cancer in Canada. Seventy two percent (74/103) of radiation oncologists who treat lung cancer from all 34 Canadian cancer centres replied to the questionnaire. RESULTS: (a) Radiotherapy regimens in Canadian cancer centres are in accordance with several major randomised studies. There is still some variation in treatment practice that may be due to unresolved controversies or limited resources. The most frequently used prescription dose was 40Gy/15f/3w (where f stands for fractions and w stands for weeks) for small cell lung cancer (SCLC) and 60Gy/30f/6w for non-small cell lung cancer (NSCLC). If there were no resource constraints, 30% (22/74) and 20% (15/74) would prefer to use a different dose-fractionation scheme for SCLC and NSCLC, respectively; 95% (70/74) would prefer to use 3D-conformal or intensity modulated radiotherapy. (b) Among the various modern technologies assessed by respondents, CT (computed tomography) simulator, multi-leaf collimator, on-line electronic portal imaging and PET (positron-emission tomography) scanning were rated the highest in terms of potential patient benefit. Discrepancy between demand and availability of technology was greatest for PET scanning. CONCLUSIONS: Canadian practice in the treatment of lung cancers shows some variations although it is consistent with the trends in the literature. The lack of some modern technologies and human resources is an ongoing concern, especially the lack of PET imaging equipment. PMID- 15110451 TI - A prospective study on radiation pneumonitis following conformal radiation therapy in non-small-cell lung cancer: clinical and dosimetric factors analysis. AB - BACKGROUND AND PURPOSE: Clinical and dosimetric prognostic factors for radiation pneumonitis (RP) have been reported after three-dimensional conformal radiotherapy (3D-CRT) in patients with non-small cell lung cancer (NSCLC). PATIENTS AND METHODS: Ninety-six patients who received 3D-CRT for stage IA to IIIB NSCLC were evaluated prospectively. Surgery was performed before radiation in 51% of the patients (n = 49). RP was diagnosed six-eight weeks after 3D-CRT using the Lent-Soma classification. Factors evaluated included treatment factors such as total mean lung dose (MLD), and dose-volume histogram (DVH) thresholds for several radiation dose steps. These thresholds were originally determined from the median of the irradiated lung volume at each step. RESULTS: Six patients could not be evaluated for RP six weeks after 3D-CRT. Of the 90 remaining patients, 40 (44%) had RP (i.e. grade > or =1) at 6 weeks, including 7 patients (7.8%) with severe RP (grade > or =2). Regarding the whole toxicity (grade > or =1), age (> or =60 years), MLD, V20 and V30 were significantly related to RP. DVH thresholds determined for radiation doses from 20 to 40 Gy were also predictive of RP. Considering only severe RP (grade > or =2), only MLD, V20 and V30 remained associated with increased acute pulmonary toxicity. CONCLUSIONS: In this study, dosimetric factors (MLD, V20, V30) and age (> or =60 years) were predictive of RP regarding the whole pulmonary toxicity (grade > or =1). In addition, thresholds from 20 to 40 Gy, based on a stratification according to the median of the percentage of irradiated lung volume, were also predictive factors. They may, therefore, help discriminate patients at high and low risk for RP. However, only MLD, V20 and V30 remained associated with severe RP (grade > or =2), probably due to the small number of severe events in our series. PMID- 15110452 TI - Transforming growth factor-beta plasma dynamics and post-irradiation lung injury in lung cancer patients. AB - PURPOSE: To investigate the relevance of transforming growth factor-beta (TGF beta) dynamics in plasma for identification of patients at low risk for developing pneumonitis as a complication of thoracic radiotherapy (RT). PATIENTS AND METHODS: Non-small cell lung cancer patients undergoing conventional RT were included in the prospective study. Concentrations of TGF-beta were measured in the patients' plasma prior to and weekly during 6 weeks of RT. The incidence of symptoms of early post-irradiation lung injury, i.e. symptomatic radiation pneumonitis, was correlated with TGF-beta parameters. RESULTS: Forty-six patients were included in the study. Eleven patients (24%) developed symptomatic radiation pneumonitis. Absolute TGF-beta plasma levels did not differ between the groups of patients without or with pneumonitis. However, patients who developed pneumonitis tended to show increases in TGF-beta levels in the middle of the RT course relative to their pre-treatment levels while TGF-beta plasma levels of patients who did not develop pneumonitis tended to decrease over the RT treatment. The difference in the relative TGF-beta dynamics between the groups reached marginal significance in the third week of the treatment (P = 0.055) but weakened towards the end of the RT course. The utility of TGF-beta testing was evaluated at each RT week based on the test's ability to yield more accurate estimate of complication probability in an individual patient compared to empirically expected probability in similar group of patients. The ratio of TGF-beta level at week 3/week 0 being <1 showed an ability to improve the prediction of freedom from pneumonitis, yet with a large degree of uncertainty (wide confidence intervals). The accuracy of prediction deteriorated at later time points (weeks 4, 5 and 6) rendering the end-RT ratios without predictive power. CONCLUSIONS: We observed a trend of plasma TGF-beta concentration to decrease below the pre treatment value during the RT treatment in patients who did not develop pulmonary complications after the RT treatment. However, this trend was not consistent enough to warrant safe decision-making in clinical setting. PMID- 15110453 TI - Dosimetric effect of respiratory motion in external beam radiotherapy of the lung. AB - BACKGROUND AND PURPOSE: To study the effect of breathing motion on gross tumor volume (GTV) coverage for lung tumors using dose-volume histograms and relevant dosimetric indices. PATIENTS AND METHODS: Treatment plans were chosen for 12 patients treated at our institution for lung carcinoma. GTV volumes of these patients ranged from 1.2 to 97.3 cm(3). A margin of 1-2 cm was used to generate the planning target volume (PTV). Additional margins of 0.6-1.0 cm were added to the PTV when designing treatment portals. For the purposes of TCP calculation, the prescription dose was assumed to be 70 Gy to remove the effects of prescription differences. Setup error was incorporated into the evaluation of treatment plans with a systematic component of sigma(RL) = 0.2 cm, sigma(AP) = 0.2 cm, and sigma(SI) = 0.3 cm and a random component of sigma(RL) = 0.3 cm, sigma(AP) = 0.3 cm, and sigma(SI) = 0.3 cm. Breathing motion was incorporated into these plans based on an independent analysis of fluoroscopic movies of the diaphragm for 7 patients. The systematic component of breathing motion (sigma(RL) = 0.3 cm, sigma(AP) = 0.2 cm, and sigma(SI) = 0.6 cm) was incorporated into the treatment plans on a slice by slice basis. The intrafractional component of breathing motion (sigma(RL) = 0.3 cm, sigma(AP) = 0.2 cm, and sigma(SI) = 0.6 cm) was incorporated by averaging the dose calculation over all displacements of the breathing cycle. Each patient was simulated 500 times to discern the range of possible outcomes. The simulations were repeated for a worst case scenario which used only breathing data with a large diaphragmatic excursion, both with and without intrafractional breathing motion. RESULTS: Dose to 95% of the GTV (D95), volume of the GTV receiving 95% of the prescription dose (V95) and TCP changed an average of -1.4+/-4.2, -1.0+/-3.3, and -1.4+/-3.8%, respectively, with the incorporation of normal breathing effects. In the worst case scenario (heavy breathers), D95 and V95 changed an average of -9.8+/-10.1 and -8.3+/-11.3%, respectively, and TCP changed by -8.1+/-9.1%. GTVs with volumes greater than 60 cm(3) showed stronger sensitivity to breathing especially if the shape was non ellipsoidal. In the normal breathing case, the probability of a decrease in D95, V95, or TCP of a magnitude greater than 10% is less than 4%, and in the worse case scenario this probability is approximately 30-40% with intrafractional breathing motion included, and less than 10% with intrafractional breathing motion not included. CONCLUSIONS: With the PTV margins routinely used at our center, the effects of normal breathing on coverage are small on the average, with a less than 4% chance of a 10% or greater decrease in D95, V95, or TCP. However, in patients with large respiration-induced motion, the effect can be significant and efforts to identify such patients are important. PMID- 15110454 TI - Target dose conformity in 3-dimensional conformal radiotherapy and intensity modulated radiotherapy. AB - BACKGROUND AND PURPOSE: Dose conformity to the planning target volume is an important criterion in radiotherapy treatment planning, for which the conformity index is a useful assessment tool. The purpose of this study is to compare the differences in CI for the treatment planning of four cancers including the nasopharynx, oesophagus, lung and prostate. PATIENTS AND METHODS: Seventy patients with cancers of nasopharynx (30), oesophagus (15), lung (15) and prostate (10) were recruited. Each of these patients was planned with three sets of treatment plans using the FOCUS treatment planning system: the forward and inverse 3DCRT plans and the IMRT plan. The CI was generated for each treatment plan. The mean CI from each cancer patient group was calculated and compared with the other three cancer groups. The mean value of CI was also compared among the three planning methods. RESULTS: The oesophageal and lung cancers demonstrated relatively higher overall mean CI values (0.64 and 0.62, respectively), whereas that of the nasopharynx and prostate were lower (0.54 and 0.50, respectively). With regards to the planning method groups, the IMRT plans produced the highest overall mean CI (0.62), while those for the forward and inverse 3DCRT were similar (0.57 and 0.55, respectively). CONCLUSION: For the four selected cancers, oesophageal and lung cancers were easier to conform than the nasopharyngeal and prostate cancers. The IMRT plans were more effective in achieving better dose conformity than that of the 3DCRT. PMID- 15110455 TI - Cell biological basis for combination radiotherapy using heavy-ion beams and high energy X-rays. AB - We investigated the biological effect of combining carbon-beam and X-ray in vitro. The results showed that when we employed Gray equivalent as the indication of therapeutic dose, the effects could be explained with simple additive way in the treatment plan. This fact provides important information about the combined therapy of carbon-beam and X-ray. PMID- 15110456 TI - Enhancement of radiation effects by combined docetaxel and flavopiridol treatment in lung cancer cells. AB - BACKGROUND AND PURPOSE: To evaluate the potential role and mechanism of docetaxel plus flavopiridol in modulating radiosensitivity in vitro and in vivo. PATIENTS AND METHODS: In vitro. H460 human lung carcinoma cells were treated with docetaxel (10 nM for 1 h, at t = 0 h) --> radiation (0-5 Gy, at t = 6 h) --> flavopiridol (120 nM for 24 h, at t = 8 h). Colony forming ability was measured to assess the modulation of sensitivity. Cell cycle redistribution was measured by flow cytometric analysis using propidium iodide. Percent apoptosis was also measured by flow cytometric analysis using 7-amino-actinomycin D staining. In vivo. H460 cell xenografts were used in nude mice. Tumors were grown subcutaneously on the flank, then treated with docetaxel (2.5 mg/kg, at t = 0 h) -> radiation (2 Gy, at t = 6 h) --> flavopiridol (1.25 mg/kg, at t = 8 h) for 5 consecutive days. Tumor growth delay was then measured and compared with the control group. RESULTS: Docetaxel plus flavopiridol enhanced the effect of radiation. The maximum radiopotentiation and apoptosis were observed when the cells were treated with the sequence of docetaxel-->radiation-->flavopiridol both in vitro and in vivo. Flavopiridol and docetaxel induced G1 and G2/M arrest, respectively. CONCLUSIONS: This study shows that docetaxel plus flavopiridol enhances the effects of radiation in vitro and in vivo. Our data suggest that the mechanism of radiopotentiation by combining flavopiridol and docetaxel involves an enhancement of apoptosis and changes of cell cycle by docetaxel and flavopiridol. PMID- 15110458 TI - A simple theoretical verification of monitor unit calculation for intensity modulated beams using dynamic mini-multileaf collimation. AB - A spreadsheet based program is presented to perform an independent Monitor Unit (MU) calculation verification for the Quality Assurance (QA) of Intensity Modulated Radiation Therapy (IMRT) using Dynamic MultiLeaf Collimation (DMLC). The computed dose value is compared to the planned dose by calculating the percent dose difference per Intensity Modulated Beam (IMB) and absolute dose difference per IMB. The proposed acceptability levels are +/-5.0% or +/-2.0 cGy for the percent dose difference per IMB and the absolute dose difference per IMB, respectively. For percent dose difference per treatment, an acceptability level of +/-2.0% is proposed. The presented program is considered adequate for checking the treatment plans calculated for IMRT treatments using DMLC as a part of the QA procedure. PMID- 15110457 TI - The use of an aSi-based EPID for routine absolute dosimetric pre-treatment verification of dynamic IMRT fields. AB - BACKGROUND AND PURPOSE: In parallel with the increased use of intensity modulated radiation treatment (IMRT) fields in radiation therapy, flat panel amorphous silicon (aSi) detectors are becoming the standard for online portal imaging at the linear accelerator. In order to minimise the workload related to the quality assurance of the IMRT fields, we have explored the possibility of using a commercially available aSi portal imager for absolute dosimetric verification of the delivery of dynamic IMRT fields. PATIENTS AND METHODS: We investigated the basic dosimetric characteristics of an aSi portal imager (aS500, Varian Medical Systems), using an acquisition mode especially developed for portal dose (PD) integration during delivery of a-static or dynamic-radiation field. Secondly, the dose calculation algorithm of a commercially available treatment planning system (Cadplan, Varian Medical Systems) was modified to allow prediction of the PD image, i.e. to compare the intended fluence distribution with the fluence distribution as actually delivered by the dynamic multileaf collimator. Absolute rather than relative dose prediction was applied. The PD image prediction was compared to the corresponding acquisition for several clinical IMRT fields by means of the gamma evaluation method. RESULTS AND CONCLUSIONS: The acquisition mode is accurate in integrating all PD over a wide range of monitor units, provided detector saturation is avoided. Although the dose deposition behaviour in the portal image detector is not equivalent to the dose to water measurements, it is reproducible and self-consistent, lending itself to quality assurance measurements. Gamma evaluations of the predicted versus measured PD distribution were within the pre-defined acceptance criteria for all clinical IMRT fields, i.e. allowing a dose difference of 3% of the local field dose in combination with a distance to agreement of 3 mm. PMID- 15110459 TI - Multiple products of the Leishmania chagasi major surface protease (MSP or GP63) gene family. AB - The major surface protease (MSP or GP63) of the Leishmania spp. protozoa facilitates parasite evasion of complement-mediated killing, phagocytosis by macrophages, and intracellular survival in macrophage phagolysosomes. Immunoblots of several Leishmania species have shown there are distinct MSP isoforms, but the biochemical bases for these differences are unknown. Northern blots show that transcripts of the three tandem gene classes encoding Leishmania chagasi MSP (MSPS, MSPL, MSPC) are differentially expressed during parasite growth in vitro. Cell-associated MSPs increase in abundance during growth, correlating directly with parasite virulence. We examined whether distinct products of these >18 MSP genes are either differentially expressed or differentially processed during parasite growth. Two-dimensional gel electrophoresis and immunoblots delineated more than 10 MSP isoforms in stationary phase L. chagasi, distributed between pIs of 5.2-6.1 and masses of 58-63 kDa. Post-translational modifications including N glycosylation, GPI anchor addition and phosphorylation did not account for all differences among the isoforms. MALDI-TOF mass spectrometry demonstrated that at least some L. chagasi MSPs were the products of different MSP genes. One isoform was not available for surface biotinylation, suggesting it could be located internally. Parasites in logarithmic growth expressed only four MSP isoforms, and an attenuated strain of L. chagasi (L5) did not express one of the MSP classes (MSPS). These data demonstrate that the products of individual MSP genes are differentially expressed during Leishmania development. We hypothesize they may play different roles during parasite migration through its two hosts. PMID- 15110460 TI - A unique Rab GTPase, EhRabA, of Entamoeba histolytica, localizes to the leading edge of motile cells. AB - Entamoeba histolytica, an enteric protozoan parasite, infects 10% of the world's population leading to 50 million cases of invasive amoebiasis annually. Parasite vesicle trafficking and motility, which relies on vesicle trafficking to deliver membrane and membrane components to the leading edge, are important for virulence however little is known about the molecular mechanisms regulating these functions. Since Rab GTPases are known modulators of vesicle trafficking we have characterized a Rab GTPase of Entamoeba, EhRabA. Sequence analysis revealed that EhRabA shared limited homology with any known Rab suggesting that it is a novel member of this protein family. Immunofluorescence microscopy using EhRabA specific antibodies demonstrated that EhRabA did not colocalize with markers for the Golgi apparatus, endoplasmic reticulum, pinosomes, or phagosomes. These data suggest that this Rab may not play a role in vesicle trafficking between these organelles. In quiescent Entamoeba cells, EhRabA localized to vesicles throughout the cytoplasm consistent with a role in vesicle trafficking, however, in motile cells this protein localized to small vesicles in the leading edge. In addition, when E. histolytica trophozoites were exposed to an N-formyl peptide (N formylmethionylleucylphenylalanine) cell polarization, the formation of membrane extensions, and the translocation of EhRabA to these membrane extensions was observed. Taken together, these results suggest that EhRabA may function in the formation of membrane extensions perhaps by regulating the delivery of membrane and/or cell surface molecules to the plasma membrane. PMID- 15110461 TI - Neutral lipid synthesis and storage in the intraerythrocytic stages of Plasmodium falciparum. AB - In eukaryotic cells the neutral lipids, steryl esters and triacylglycerol, are synthesized by membrane-bound O-acyltransferases and stored in cytosolic lipid bodies. We show here that the intraerythrocytic stages of Plasmodium falciparum produce triacylglycerol using oleate and diacylglycerol as substrates. Parasite membrane preparations reveal a synthesis rate of 4.5 +/- 0.8 pmol x min(-1)mg(-1) of protein with maximal production occurring in the mid- and late-trophozoite stages in both, membrane preparations and live parasites. In contrast to other eukaryotic cells, no discernable amounts of steryl esters are produced, and the parasite is insensitive to cholesterol esterification inhibitors. Synthesized neutral lipids are stored as lipid bodies in the parasite cytosol in a stage specific manner. Their biogenesis is not modified upon incubation with excess fatty acids or lipoproteins or after lipoprotein depletion of the culture medium. We investigated on the enzymes involved in neutral lipid synthesis and found that only one gene with significant homology to known members of the membrane-bound O acyltransferase family is present in the P. falciparum genome. It encodes a microsomal transmembrane protein with a predicted size of 78.1 kDa, which we named PfDGAT because of its close identity with various known acyl CoA:diacylglycerol acyltransferases. PfDGAT is expressed in a stage specific manner as documented by Western blotting and immunoprecipitation assays using antibodies against Toxoplasma DGAT, suggesting that PfDGAT is the most likely candidate for plasmodial triacylglycerol synthesis. PMID- 15110462 TI - Purification and properties of poly(ADP-ribose)polymerase from Crithidia fasciculata. Automodification and poly(ADP-ribosyl)ation of DNA topoisomerase I. AB - Poly(ADP-ribose)polymerase has been purified more than 160000-fold from Crithidia fasciculata. This is the first PARP isolated to apparent homogeneity from trypanosomatids. The purified enzyme absolutely required DNA for catalytic activity and histones enhanced it 2.5-fold, when the DNA:histone ratio was 1:1.3. The enzyme required no magnesium or any other metal ion cofactor. The apparent molecular mass of 111 kDa, determined by gel filtration would correspond to a dimer of two identical 55-kDa subunits. Activity was inhibited by nicotinamide, 3 aminobenzamide, theophylline, thymidine, xanthine and hypoxanthine but not by adenosine. The enzyme was localized to the cell nucleus. Our findings suggest that covalent poly(ADP-ribosyl)ation of PARP itself or DNA topoisomerase I resulted in the inhibition of their activities and provide an initial biochemical characterization of this covalent post-translational modification in trypanosomatids. PMID- 15110463 TI - The I-complex in Leishmania tarentolae is an uniquely-structured F(1)-ATPase. PMID- 15110464 TI - The mitochondrial ribosome of the protozoan Acanthamoeba castellanii is the target for macrolide antibiotics. PMID- 15110465 TI - Responses of benthic fish exposed to contaminants in outdoor microcosms- examining the ecological relevance of previous laboratory toxicity tests. AB - Previous laboratory studies indicate that coal combustion wastes (a mixture composed of fly ash and other lower volume wastes such as bottom ash; hereafter collectively referred to as ash) adversely affect the health of benthic fish (Erimyzon sucetta; lake chubsucker), but fish in these studies were provided with ample uncontaminated food resources. Because aquatic disposal of ash can also adversely affect food resources for benthic fish, we hypothesized that changes in resources might exacerbate the effects of ash on fish observed in laboratory studies. We exposed juvenile E. sucetta in outdoor microcosms to water, sediment, and benthic resources from an ash-contaminated site or a reference site for 45 days and compared our findings to previous laboratory studies. Benthic invertebrate biomass was nearly three times greater in controls compared to ash microcosms. Total organic content of control sediment (41%) was also greater than in ash sediments (17%), suggesting that additional benthic resources may have also been limited in ash microcosms. Benthic invertebrates isolated from the ash microcosms had trace element concentrations (As, Cd, Co, Cr, Cs, Se, Sr, and V) up to 18 times higher than in weathered ash used in laboratory studies. The concentrations of trace elements accumulated by fish reflected the high dietary concentrations encountered in the ash microcosms and were associated with reduced growth (final mass = 0.07 g) and survival (25%) compared to controls (0.37 g and 67%, respectively). Accumulation of trace elements, as well as reductions in growth and survival, were more pronounced than in previous laboratory studies, suggesting that resource conditions may be important in mediating ash toxicity. Taken together, our studies suggest that ash discharge into aquatic systems is a more serious threat to the health of benthic fish than previously predicted based upon laboratory toxicity tests. PMID- 15110467 TI - On the relevance of genotoxicity for fish populations II: genotoxic effects in zebrafish (Danio rerio) exposed to 4-nitroquinoline-1-oxide in a complete life cycle test. AB - In order to characterize the impact of genotoxic potentials on populations of aquatic organisms in surface waters, zebrafish (Danio rerio) were exposed to the model genotoxicant 4-nitroquinoline-1-oxide (NQO) in a complete life-cycle test. Fish exposed to mean NQO concentrations of 0, 0.1, 0.3, 1.1, and 2.9 microg/l were examined by several genotoxicity assays with different endpoints. Assays included the unscheduled DNA synthesis (UDS) test, the comet assay, the alkaline filter elution, and the micronucleus test. The genotoxicity assays revealed an increasing genotoxicity, ranging from induction of DNA repair (even at the lowest concentration tested) to primary and secondary DNA alterations at higher concentrations of 1.1 and 2.9 microg/l NQO. Whether the lowered reproductivity observed in the life-cycle test is caused by genotoxic pathways of NQO, remains unclear. However, the results indicate a contradiction to an earlier assumption that genotoxicants as found in the environment are likely to not impact natural populations. PMID- 15110466 TI - On the relevance of genotoxicity for fish populations I: effects of a model genotoxicant on zebrafish (Danio rerio) in a complete life-cycle test. AB - Genotoxicity may be detected in surface waters by means of various genotoxicity assays. In order to enable an ecotoxicological assessment of the consequences of such genotoxic potential for fish populations, a complete life-cycle test with zebrafish (Danio rerio) and the model genotoxicant 4-nitroquinoline-1-oxide (NQO) was conducted. Zebrafish (f1) were continuously exposed to NQO (i.e. 0.1, 0.3, 1.1, 2.9, and 14.6 microg/l, respectively) from fertilised eggs until sexual maturity. In addition to reproduction studies in the f1-generation, f2-fish were exposed to NQO during the first 6 weeks of development. Up to 2.9 microg/l NQO, fish did not display differences in survival and growth (P < 0.05). A NQO concentration of 14.6 microg/l, however, was lethal. Female fish exposed to all NQO concentrations up to 2.9 microg/l displayed a significant reduction in egg production (P < 0.05). A mathematical simulation revealed that exposure to weak concentrations of NQO is leading to an elevated extinction risk. PMID- 15110468 TI - Influences of different selenium species on the uptake and assimilation of Hg(II) and methylmercury by diatoms and green mussels. AB - We examined the influences of different concentrations and species of Se (selenite, selenate, and seleno-l-methionine) in the ambient environment on the accumulation of inorganic Hg(II) and methylmercury (MeHg) by the diatom Thalassiosira pseudonana and the green mussel Perna viridis. At the experimental concentrations tested (< 500 microgl(-1)), selenite and selenate did not significantly affect the uptake of either mercury species by the diatoms and the green mussels. The assimilation efficiency of Hg(II) and MeHg by the mussels from ingested diatoms was also independent of the inorganic Se loadings in the food particles. In contrast, selenomethionine significantly inhibited the uptake of MeHg and enhanced the uptake of Hg(II) by the diatoms and the mussels, but it did not affect the assimilation from the ingested diatoms. The influence of tissue body burden of Se in the green mussels following pre-exposure to selenite and selenomethionine for different periods (1-5 weeks) on the accumulation of Hg(II) and MeHg was further investigated. Our results showed that tissue Se concentrations did not significantly affect the dietary assimilation of mercury, but the influences on the aqueous uptake were variable. Our study thus, strongly highlights the specificity of the Se-Hg interaction in marine mussels for different Se and Hg species. Both dissolved and dietary uptake appeared to be equally important in the accumulation of Hg(II) and MeHg in the green mussels. PMID- 15110469 TI - Changes in microcystin production by Microcystis aeruginosa exposed to phytoplanktivorous and omnivorous fish. AB - With direct exposure to phytoplanktivorous fish (Hypophthalmichthys molitrix), increased mass-specific microcystin production occurred in three monoclonal Microcystis aeruginosa strains (NIES 44, 88 and 99). Total mass-specific microcystin content of NIES 44 exposed to H. molitrix was over 50 times higher than controls (a mean value of 16.2 microgg(-1)-dry cell in controls versus 878.6 microgg(-1)-dry cell in treatments). Up to nine times higher microcystin levels were detected in NIES 88 exposed to H. molitrix compared to controls (a mean value of 553 in controls versus 5145 microgg(-1)-dry cell in treatments). The microcystin levels of all strains were significantly different between controls and H. molitrix treatments (P < 0.01 for NIES 44 and 88; P < 0.05 for NIES 99). The microcystin response to the omnivorous Carassius gibelio langsdorfi was weaker than that of H. molitrix, though the levels in all strains exposed to the fish were higher than in controls and a significant difference in microcystin production between controls and omnivorous fish treatments occurred for NIES 44 (a mean value of 6.9 in controls versus 41.5 microgg(-1)-dry cell in treatments; P < 0.01) and NIES 88 (a mean value of 359.8 versus 480.4 microgg(-1)-dry cell; P < 0.05). Microcystis cells were observed in the both fish faeces and gut contents, and microcystin was also detected in the body tissues (from 0.6 to 2.5 microgg(-1)-dry weight) and faeces of both fish species on the final day of experiment, although 98% of fish in three strains of Microcystis cultures had lost weight (mean +/- S.E. fish growth rate with M. aeruginosa; -0.90 +/- 0.06% per day, n = 96). This study showed that several M. aeruginosa strains increased toxin production when exposed to fish, especially phytoplanktivorous species, even though fish appeared not to feed vigorously on toxic Microcystis, and supports the hypothesis that this response is a fish-induced defence mediated by physical contact associated with feeding or by chemical cues (e.g. kairomones). PMID- 15110470 TI - Long-term acclimation of Pseudokirchneriella subcapitata (Korshikov) Hindak to different copper concentrations: changes in tolerance and physiology. AB - The effect of long-term copper acclimation of the freshwater green algae Pseudokirchneriella subcapitata to copper was investigated using different physiological and toxicological endpoints. The algae were exposed to seven-five of which are ecologically relevant for European surface waters-copper concentration ranging from 0.5 to 100 microgCul(-1) during a 3-month period. A standard medium was used as culture and test medium with an addition of 2 mg DOCl(-1) (replacing EDTA). At certain intervals, experiments were performed to assess algal biomass, growth rate, chlorophyll and carotenoid content, pigment diversity, autotrophic index, intracellular and adsorbed copper, and the sensitivity of the algae to copper. Chronic copper tolerance (mean +/- standard deviation) increased significantly from 88 +/- 15 to 124 +/- 25 microg Cul(-1) for P. subcapitata acclimated to 0.5 and 100 microg Cul(-1), respectively. Based on the algal biomass, the growth rate, the pigment diversity and the autotrophic index, an optimal concentration range was observed between 1 and 35 microg Cul( 1). Significant decreases in algal biomass, pigment diversity and autotrophic index were observed in algal cultures acclimated to 0.5 microg Cul(-1) and 100 microg Cul(-1). Chlorophyll a content (mean +/- standard deviation) increased from 8.4 +/- 3.1 to 28.6 +/- 7.5 x 10(-14) g per cell and carotenoid content (mean +/- standard deviation) increased from 3.7 +/- 0.8 to 7.1 +/- 1.2 x 10(-14) g per cell for algae exposed to 1 and 100 microg Cul(-1), respectively. Intracellular copper increased from 0.099 to 20.6 x 10(-15) g Cu per cell and adsorbed copper increased from 0.026 to 1.8 x 10(-15) g Cu per cell for algae acclimated for 12 weeks to 0.5 and 100 microg Cul(-1), respectively. This research demonstrates that the use of standard culture media, some of which may be deficient in copper, can result in sub-optimal performance of the organisms, which in turn may affect toxicity test results. Additionally, this work also established an optimal concentration range for copper for this algal species. This phenomenon should be taken in consideration when performing environmental risk assessments of essential elements. PMID- 15110471 TI - Pentachlorophenol (PCP) bioaccumulation and effect on heat production on salmon eggs at different stages of development. AB - In this study, pentachlorophenol (PCP) bioaccumulation and its effect on heat dissipation was studied in eggs of the lake salmon (Salmo salar m. sebago). In bioaccumulation studies, the eggs were exposed to low concentrations (0.051-0.056 micromol/l, 13.583-14.915) of waterborne [14C]-labeled PCP at two developmental stages: (1) 3 weeks after fertilization, and (2) just before hatching. The effect of PCP on egg heat dissipation was measured by a microcalorimeter after exposing the eggs to gradual concentrations (0-0.992 micromol/l) of PCP for 48 h. After both the bioaccumulation and heat dissipation experiments, the eggs were dissected and the concentrations of PCP in tissue were determined separately for eggshell, yolk and embryo. The bioaccumulation studies showed that PCP accumulates more in the eggs at the late developmental stage. Bioconcentration factors (BCF) for different tissues were 3-42 times higher for the eggs at the late developmental stage compared with the eggs that were incubated only for 3 weeks. In early developmental stage, the eggshell adsorbs a large portion of the chemical. In late developmental stage, the actual embryo accumulated both proportionately and totally more than other dissected tissues in the beginning of the exposure, but eventually the yolk accumulated highest total amount of the chemical. A probable reason for the higher PCP body burden in the late developmental stage is that the respiration rate and metabolic activity of the embryo increases as it grows. The salmon eggs responded to an exposure to PCP with an elevated rate of heat dissipation. The threshold concentration above which the embryo heat dissipation was amplified was 29.64 micromol/kg embryo wet weight (ww) or 0.28 micromol/l. The highest embryo heat production was measured at the exposure concentration of 0.992 micromol/l. At higher exposure concentrations the heat dissipation decreased. The basic findings of the study are that PCP accumulates in growing embryonic tissue and is able to change the physiology of developing embryo. PMID- 15110473 TI - Toughening of dental glass ionomer cements with reactive glass fibres. AB - A fibre reinforced glass ionomer cement (FRGIC) for dental applications was loaded with 20 vol% short fibres (430 microm) with a glass composition in the system SiO(2)-Al(2)O(3)-CaF(2)-Na(3)AlF(6). The fracture toughness and the total energy release rate were examined. A 20% anisotropic fibre alignment was observed, perpendicular to the loading direction. An increase of fracture toughness of 140% and of total energy release rate of 440% was achieved compared to the unreinforced glass ionomer cement. Matrix-fibre interface reaction is supposed to exert the major influence on mechanical behaviour of FRGIC by controlling fibre pull-out and thus the total energy release rate. PMID- 15110472 TI - Effect of three xenobiotic compounds on Glutathione S-Transferase in the clam Ruditapes decussatus. AB - The effects of 4,4'DDE, methoxychlor and imidazole were studied on glutathione S transferase activities in the gills and hepatopancreas of the clam Ruditapes decussatus. The contamination doses were 0.14 microM for 4,4'DDE, 0.14 microM for methoxychlor and 25 microM for imidazole. GST activities were spectrophotometrically measured. SDS-PAGE and isoelectric focusing (IEF) were used to separate the different GST isoforms in control and treated animals, followed by Western blotting performed with anti-alpha, anti-mu and anti-pi GST anti-sera. In the hepatopancreas, GST-CDNB activities were always two to five fold lower than in the gills where the activities were significantly increased after exposure to 4,4'DDE (ca. 1.6-fold) and to methoxychlor (ca. 1.3-fold) compared to the controls (ca. 3 micromolmin(-1)mg(-1)protein) whereas they remained unchanged after treatment with imidazole. When using glutathione S transferase anti-alpha, anti-mu and anti-pi anti-sera, a single 26 kDa polypeptide was observed in the hepatopancreas and in the gills in all the tested conditions. Five GST subunits were observed after IEF showing greater immuno reactivity with the anti-pi mammalian class antiserum than with the anti-alpha or anti-mu mammalian anti-sera. One isoform of pI 5.77 was particularly induced by 4,4'DDE and methoxychlor; it was recognized by the three anti-sera tested and seemed to be more efficient in the gills than in the hepatopancreas. This isoform may play a role in organochlorine detoxication. PMID- 15110474 TI - Novel porous aortic elastin and collagen scaffolds for tissue engineering. AB - Decellularized vascular matrices are used as scaffolds in cardiovascular tissue engineering because they retain their natural biological composition and three dimensional (3-D) architecture suitable for cell adhesion and proliferation. However, cell infiltration and subsequent repopulation of these scaffolds was shown to be unsatisfactory due to their dense collagen and elastic fiber networks. In an attempt to create more porous structures for cell repopulation, we selectively removed matrix components from decellularized porcine aorta to obtain two types of scaffolds, namely elastin and collagen scaffolds. Histology and scanning electron microscopy examination of the two scaffolds revealed a well oriented porous decellularized structure that maintained natural architecture of the aorta. Quantitative DNA analysis confirmed that both scaffolds were completely decellularized. Stress-strain analysis demonstrated adequate mechanical properties for both elastin and collagen scaffolds. In vitro enzyme digestion of the scaffolds suggested that they were highly biodegradable. Furthermore, the biodegradability of collagen scaffolds could be controlled by crosslinking with carbodiimides. Cell culture studies showed that fibroblasts adhered to and proliferated on the scaffold surfaces with excellent cell viability. Fibroblasts infiltrated about 120 microm into elastin scaffolds and about 40 microm into collagen scaffolds after 4 weeks of rotary cell culture. These results indicated that our novel aortic elastin and collagen matrices have the potential to serve as scaffolds for cardiovascular tissue engineering. PMID- 15110475 TI - Synthesis and cell affinity of functionalized poly(L-lactide-co-beta-malic acid) with high molecular weight. AB - A novel functionalized biodegradable poly(L-lactide-co-beta-benzyl malolactonate) (p-PLMA) with high molecular weight was synthesized through ring-opening copolymerization. Three p-PLMA copolymers with different beta-benzyl malolactonate content were synthesized. The molecular weight (M(w)) and tensile strength of the copolymer with 4 mol% beta-benzyl malolactonate content were 179,800 and 19.0MPa respectively, the molecular weight (M(w)) and tensile strength of p-PLMA decreased with beta-benzyl malolactonate content increasing. The hydrophilicity of the de-protected product: poly(L-lactide-co-beta-malic acid) (d-PLMA) increased with malic acid content increasing. The results of 3T3 mice fibroblasts cultivated on d-PLMA films showed that the cell adhesion on d PLMA was better than that of PLLA and the cell attached efficiency of d-PLMA with 8 mol% malic acid content was the highest. The cells grew well both on the surface and inside of d-PLMA scaffolds. The cell affinity of d-PLMA was better than that of PLLA. PMID- 15110476 TI - Poly(2-hydroxyethyl methacrylate)-based slabs as a mouse embryonic stem cell support. AB - Poly(2-hydroxyethyl methacrylate) (PHEMA) crosslinked with ethylene dimethacrylate (EDMA) or N,O-dimethacryloylhydroxylamine (DMHA) was obtained in the form of slabs by bulk radical polymerization. Two porosity-inducing methods were investigated, phase separation using a low-molecular-weight porogen and a salt-leaching technique using NaCl and saccharose. Compared with the phase separation, the salt-leaching created open porous structures with voids of the size and shape of crystallites. To address its potentials in the context of stem cell therapies, undifferentiated mouse embryonic stem cells D3 (ES D3 cells) were seeded on the slabs and analyzed for the ability to grow on different types of non-degradable and/or degradable porous PHEMA hydrogels. The cells were able to proliferate only on PHEMA crosslinked with EDMA or 2 wt% DMHA. In order to assess the effect of gelatin, which is routinely used for ES cell cultures, PHEMA slabs were soaked in gelatin solutions and compared the number of cells on gelatin treated and untreated slabs 4 days after cell seeding. Surprisingly, the number of cells was only slightly higher on gelatin-treated slabs. PMID- 15110477 TI - Synthesis, characterization and in vitro degradation of a biodegradable elastomer. AB - An elastomer was prepared from biodegradable components as a potential biomaterial for drug delivery and tissue engineering applications. The elastomer was synthesized in two steps. First, a star copolymer (SCP) was manufactured via ring opening polymerization of -caprolactone (epsilon-CL) with D,L-lactide using glycerol as initiator and stannous 2-ethylhexanoate as catalyst. This living SCP was further reacted with different ratios of a crosslinking monomer, 2,2 bis(epsilon-CL-4-yl)-propane in the presence of epsilon-CL as a solvent and co monomer. The elastomers had very low glass transitions (-32 degrees C), sol contents ranging from 17% to 37%, and were soft and weak with physical properties similar to those of natural elastomers such as elastin. The physical properties decreased in a logarithmic fashion with time when degraded in phosphate buffered saline, indicative of first-order degradation kinetics. The elastomers degraded relatively slowly, with degradation being incomplete after 12 weeks. PMID- 15110479 TI - Temperature and pH-responsive polymeric composite membranes for controlled delivery of proteins and peptides. AB - This work was focused on the investigation of temperature and pH-responsive polymeric composite membranes and their permeability to proteins and peptides in response to environmental stimuli. The composite membranes were prepared from nanoparticles of poly(N-isopropylacrylamide-co-methacrylic acid) of various NIPAAm:MAA ratios dispersed in a matrix of a hydrophobic polymer. N-Benzoyl-L tyrosine ethyl ester HCl, momany peptide, Leuprolide, vitamin B(12), insulin, and lysozyme were used as model solutes. The morphology of the membranes was examined with SEM and permeation of the solutes was measured using side-by-side diffusion cells at varied temperatures and pH. Permeability of the solutes across the membranes increased with increasing temperature or particle concentration, while decreased with increasing pH and molecular size of the solutes. Membranes containing nanoparticles of more NIPAAm units exhibited higher thermal sensitivity, and those with higher MAA content showed more pH responsiveness, which was in line with the temperature and pH-responsive volume change of the nanoparticles. The change in permeability was quickly detected following the application of the stimuli. These results and partition study using vitamin B(12) supported the proposed gel-pore mechanism of solute permeation through these composite membranes. PMID- 15110478 TI - Oligo-3-hydroxybutyrates as potential carriers for drug delivery. AB - In the present paper we describe the synthesis and toxicity studies of well defined tailor made oligo-[R,S]-3-hydroxybutyrates (OHBs). The results indicate potential applicability of these nano-polymers as drug delivery carriers. Several OHBs of number average molecular weight (M(n)) ranging from 800 to 2400 have been synthesized and tested on transformed hamster V79 fibroblasts and murine melanoma B16(F10) cells using the 3-[4,5-dimethylthiazol-2-yl]2,5-diphenyltetrazolium bromide (MTT) based drug resistance and clonogenic survival assays. We show that 96-h incubation of cells with 1-9 microg/ml of OHBs did not affect cell viability. Incubation of OHBs with rat hepatoma FTO-2B cells stably transfected with chloramphenicol acetyltransferase (CAT) gene ligated to heat-inducible hsp70i gene promoter demonstrated that OHBs did not induce cellular stress response. Finally, we demonstrate that doxorubicin conjugated with OHB is effectively taken up by murine melanoma B16(F10) cells in vitro and localizes in the cytoplasm. These data show for the first time that tailor-made biodegradable and biocompatible oligomers of 3-hydroxybutyric acid can be taken into consideration as effective, non-toxic vectors for delivery of drugs in a conjugated form. PMID- 15110480 TI - The effect of the degree of chitosan deacetylation on the efficiency of gene transfection. AB - Chitosans with various degrees of deacetylation were synthesized by acetylation with acetic anhydride. These chitosans were evaluated for efficacy of nanoparticle formation, DNA binding efficiency, morphology, and in vitro and in vivo gene transfection efficiency. DNA binding efficacy was reduced as degree of deacetylation was decreased, therefore requiring an increased +/-ratio to effect complete DNA complexation. For chitosan with a molecular weight of 390 kDa, the +/-ratio to achieve complete DNA complexation for degrees of deacetylation of 90%, 70% and 62% was 3.3:1, 5.0:1, and 9.0:1, respectively. The size and morphology of these nanoparticle formulations were not significantly different. The decreased degree of deacetylation results in a decrease in overall luciferase expression levels in HEK293, HeLa, and SW756 cells due to particle destabilization in the presence of serum proteins. However, intramuscular luciferase expression levels increased with decreasing deacetylation over the time points tested. Degree of chitosan deacetylation is an important factor in chitosan-DNA nanoparticle formulation as it affects DNA binding, release and gene transfection efficiency in vitro and in vivo. PMID- 15110481 TI - Calcium phosphate sol-gel-derived thin films on porous-surfaced implants for enhanced osteoconductivity. Part I: Synthesis and characterization. AB - Thin sol-gel-formed calcium phosphate (Ca-P) films were formed on sintered porous surfaced implants as an approach to increasing the rate of bone ingrowth. The films were prepared using either an inorganic precursor solution (with calcium nitrate tetrahydrate and ammonium dihydrogen phosphate) or an organic precursor solution (with calcium nitrate tetrahydrate and triethyl phosphite). We report on the formation and characteristics of the films so formed. Film characteristics were assessed by thin film X-ray diffraction, diffuse-reflectance infrared Fourier transform spectroscopy, X-ray photoelectron spectroscopy, and scanning electron microscopy. In addition, thin sections were prepared either across or parallel to the Ca-P/Ti6Al4V interface and examined by transmission electron microscopy. Both approaches resulted in the formation of nanocrystalline carbonated hydroxyapatite films but with different Ca/P ratios and structures, the Inorganic Route-formed film having a lower Ca/P ratio (1.46 cf 2.10 for the Organic Route-formed film) and having a more irregular topography. An interfacial reaction product (CaTi(2)O(5)) was identified by selected area electron diffraction with the Inorganic Route-formed film only. PMID- 15110482 TI - Calcium phosphate sol-gel-derived thin films on porous-surfaced implants for enhanced osteoconductivity. Part II: Short-term in vivo studies. AB - Osseointegration rates of porous-surfaced Ti6Al4V implants with control (unmodified sintered coatings) were compared to porous-surfaced implants modified through the addition of either an Inorganic or Organic Route-formed-Ca-P film. Implants were placed in distal femoral rabbit condyle sites and, following a 9 day healing period, implant fixation strength was evaluated using a pull-out test. Three groups of ten rabbits each were evaluated. Inorganic Route Ca-P coated implants were compared with control implants in Group I. Organic Route Ca P-coated implants with control implants in Group II, and Inorganic- with Organic Route-Ca-P-coated implants in Group III. Maximum pull-out force and interface stiffness were compared while selected extracted implants were examined by SEM to characterise failure surfaces. Both types of Ca-P coatings significantly enhanced the early rate of bone ingrowth and fixation as evidenced by higher pull-out force and interface stiffness compared with controls. However, there was no significant difference between Ca-P-coated implants prepared using the two different methods. The enhanced osteoconductivity observed with the Organic Route formed films despite the absence of any obvious new surface topographic features introduced with the films suggests that the increased rate of bone ingrowth was due primarily to altered surface chemistry rather than changes in topography, at least for these sintered porous-surfaced implants. PMID- 15110483 TI - The effect of pH on the structural evolution of accelerated biomimetic apatite. AB - The classic biomimetic apatite coating process can be accelerated by first immersing substrates into concentrated simulated body fluid, 5x SBF (SBF1), at 37 degrees C, to form an initial coating of precursor apatite spheres, and subsequently transferring to a second 5x SBF (SBF2) solution which is devoid of crystal growth inhibitors to promote phase transformation of SBF1-derived precursor apatite spheres into final crystalline apatite plates. Since SBF1 governs the formation kinetics and composition of the initial precursor spheres, we hypothesized that the pH of the SBF1 solution will also influence the final structure of the SBF2-derived crystalline apatite. To test this hypothesis, polystyrene substrates were immersed into SBF1 with different pH (5.8 or 6.5), and then immersed into the identical SBF2 (pH=6.0). The resultant apatites exhibited similar 2 theta XRD peaks; FTIR spectra in terms of hydroxyl, phosphate and carbonate groups; and Ca/P atomic ratio (1.42 for SBF1(5.8) apatite; 1.48 for SBF1(6.5) apatite). SEM, TEM and electron diffraction show that while SBF1(6.5) (pH 6.5) precursor spheres transform into larger, single crystals plates, SBF1(5.8) (pH 5.8) precursor spheres developed minute, polycrystalline plate-like structures over predominantly spherical precursor substrate. PMID- 15110484 TI - Depressing research. PMID- 15110485 TI - The growth hormone and insulin-like growth factor I axis, and cancer. PMID- 15110486 TI - Functional genomics and proteomics for infectious diseases in the post-genomics era. PMID- 15110487 TI - Spatial confinement of isoforms of cardiac nitric-oxide synthase: unravelling the complexities of nitric oxide's cardiobiology. PMID- 15110488 TI - Communicable disease surveillance and management in a globalised world. PMID- 15110489 TI - Pilates to pit. PMID- 15110490 TI - Selective serotonin reuptake inhibitors in childhood depression: systematic review of published versus unpublished data. AB - BACKGROUND: Questions concerning the safety of selective serotonin reuptake inhibitors (SSRIs) in the treatment of depression in children led us to compare and contrast published and unpublished data on the risks and benefits of these drugs. METHODS: We did a meta-analysis of data from randomised controlled trials that evaluated an SSRI versus placebo in participants aged 5-18 years and that were published in a peer-reviewed journal or were unpublished and included in a review by the Committee on Safety of Medicines. The following outcomes were included: remission, response to treatment, depressive symptom scores, serious adverse events, suicide-related behaviours, and discontinuation of treatment because of adverse events. FINDINGS: Data for two published trials suggest that fluoxetine has a favourable risk-benefit profile, and unpublished data lend support to this finding. Published results from one trial of paroxetine and two trials of sertraline suggest equivocal or weak positive risk-benefit profiles. However, in both cases, addition of unpublished data indicates that risks outweigh benefits. Data from unpublished trials of citalopram and venlafaxine show unfavourable risk-benefit profiles. INTERPRETATION: Published data suggest a favourable risk-benefit profile for some SSRIs; however, addition of unpublished data indicates that risks could outweigh benefits of these drugs (except fluoxetine) to treat depression in children and young people. Clinical guideline development and clinical decisions about treatment are largely dependent on an evidence base published in peer-reviewed journals. Non-publication of trials, for whatever reason, or the omission of important data from published trials, can lead to erroneous recommendations for treatment. Greater openness and transparency with respect to all intervention studies is needed. PMID- 15110491 TI - Insulin-like growth factor (IGF)-I, IGF binding protein-3, and cancer risk: systematic review and meta-regression analysis. AB - BACKGROUND: Insulin-like growth factor (IGF)-I and its main binding protein, IGFBP-3, modulate cell growth and survival, and are thought to be important in tumour development. Circulating concentrations of IGF-I might be associated with an increased risk of cancer, whereas IGFBP-3 concentrations could be associated with a decreased cancer risk. METHODS: We did a systematic review and meta regression analysis of case-control studies, including studies nested in cohorts, of the association between concentrations of IGF-I and IGFBP-3 and prostate, colorectal, premenopausal and postmenopausal breast, and lung cancer. Study specific dose-response slopes were obtained by relating the natural log of odds ratios for different exposure levels to blood concentrations normalised to a percentile scale. FINDINGS: We identified 21 eligible studies (26 datasets), which included 3609 cases and 7137 controls. High concentrations of IGF-I were associated with an increased risk of prostate cancer (odds ratio comparing 75th with 25th percentile 1.49, 95% CI 1.14-1.95) and premenopausal breast cancer (1.65, 1.26-2.08) and high concentrations of IGFBP-3 were associated with increased risk of premenopausal breast cancer (1.51, 1.01-2.27). Associations were larger in assessments of plasma samples than in serum samples, and in standard case-control studies compared with nested studies. INTERPRETATION: Circulating concentrations of IGF-I and IGFBP-3 are associated with an increased risk of common cancers, but associations are modest and vary between sites. Although laboratory methods need to be standardised, these epidemiological observations could have major implications for assessment of risk and prevention of cancer. PMID- 15110492 TI - Seamless management of biliary atresia in England and Wales (1999-2002). AB - BACKGROUND: Before 1999, infants born in the UK with suspected biliary atresia were investigated in regional centres, and, if confirmed, a Kasai operation was done there. Since 1999, all infants with suspected biliary atresia in England and Wales, UK, have been referred to one of three designated centres where both the Kasai operation and liver transplantation (if necessary) could be done. METHODS: We assessed clearance of jaundice (bilirubin <20 micromol/L) as an early outcome in all cases of biliary atresia referred from one of the three centres. We then estimated survival using the Kaplan-Meier method with endpoints of liver transplantation or death. FINDINGS: 148 infants with biliary atresia were treated between January, 1999, and June, 2002. A primary portoenterostomy was done in 142 (96%) infants and a primary liver transplant in five (3%). One child died before any intervention. Early clearance of jaundice after portoenterostomy was achieved in 81 of 142 (57%) infants. Liver transplantation was done in 52 (37%) of those undergoing portoenterostomy. 13 (9%) infants died. Of the 135 children who survived, 84 (62%) still have their native liver and 51 (38%) had transplantation. The median follow-up of survivors was 2.13 (range 0.5-4.1) years. The overall 4-year estimated actuarial survival was 89% (95% CI 82-94). The 4-year estimated actuarial survival with native liver was 51% (42-59%). INTERPRETATION: Our early results suggest that surgical outcome can be improved by centralisation of care to supra-regional centres. PMID- 15110493 TI - A novel and accurate diagnostic test for human African trypanosomiasis. AB - INTRODUCTION: Human African trypanosomiasis (sleeping sickness) affects up to half a million people every year in sub-Saharan Africa. Because current diagnostic tests for the disease have low accuracy, we sought to develop a novel test that can diagnose human African trypanosomiasis with high sensitivity and specificity. METHODS: We applied serum samples from 85 patients with African trypanosomiasis and 146 control patients who had other parasitic and non parasitic infections to a weak cation exchange chip, and analysed with surface enhanced laser desorption-ionisation time-of-flight mass spectrometry. Mass spectra were then assessed with three powerful data-mining tools: a tree classifier, a neural network, and a genetic algorithm. FINDINGS: Spectra (2-100 kDa) were grouped into training (n=122) and testing (n=109) sets. The training set enabled data-mining software to identify distinct serum proteomic signatures characteristic of human African trypanosomiasis among 206 protein clusters. Sensitivity and specificity, determined with the testing set, were 100% and 98.6%, respectively, when the majority opinion of the three algorithms was considered. This novel approach is much more accurate than any other diagnostic test. INTERPRETATION: Our report of the accurate diagnosis of an infection by use of proteomic signature analysis could form the basis for diagnostic tests for the disease, monitoring of response to treatment, and for improving the accuracy of patient recruitment in large-scale epidemiological studies. PMID- 15110494 TI - A paralysed thumb. PMID- 15110495 TI - Increased neuronal nitric oxide synthase-derived NO production in the failing human heart. AB - Experimental data suggest that nitric oxide (NO) generated from neuronal NO synthase (nNOS) modulates the myocardial inotropic state. To assess the contribution of NO, derived from endothelial and neuronal isoforms, to the pathophysiology of congestive heart failure in human beings, we compared expression, localisation, and specific activity of NOS isoforms in myocardium from patients with dilated cardiomyopathy with those in controls who had died from head trauma or intracranial bleeds. Diseased hearts had a significant increase in nNOS mRNA and protein expression, and activity associated with the translocation of nNOS to the sarcolemma through interactions with caveolin 3. Enhanced nNOS activity counteracted a decrease in eNOS expression and activity. Our results provide evidence of increased nNOS-derived NO in the failing human heart. Such altered regulation may be important in the pathophysiology of cardiac dysfunction in human congestive heart failure. PMID- 15110496 TI - Errors based on units of measure. PMID- 15110498 TI - Osteogenesis imperfecta. AB - Osteogenesis imperfecta is a genetic disorder of increased bone fragility, low bone mass, and other connective-tissue manifestations. The most frequently used classification outlines four clinical types, which we have expanded to seven distinct types. In most patients the disorder is caused by mutations in one of the two genes encoding collagen type 1, but in some individuals no such mutations are detectable. The most important therapeutic advance is the introduction of bisphosphonate treatment for moderate to severe forms of osteogenesis imperfecta. However, at present, the best treatment regimen and the long-term outcomes of bisphosphonate therapy are unknown. Although this treatment does not constitute a cure, it is an adjunct to physiotherapy, rehabilitation, and orthopaedic care. Gene-based therapy presently remains in the early stages of preclinical research. PMID- 15110499 TI - Malaria intermittent preventive treatment in infants, chemoprophylaxis, and childhood vaccinations. AB - CONTEXT: Malaria accounts for 1-3 million deaths yearly worldwide, mostly in children under 5 years of age in sub-Saharan Africa. Laboratory and clinical studies show an association between acute malaria and a decreased response to diphtheria and tetanus toxoids and to meningococcal, salmonella, and Haemophilus influenzae type b vaccinations. Malaria treatment, chemoprophylaxis, or other forms of parasite suppression might improve the immune response to childhood vaccinations. However, the antimalarial 4-aminoquinolones are immunodepressive, such that antimalarial drugs might depress the vaccine response. STARTING POINT: Last year, Julius Massaga and colleagues reported a randomised double-blinded placebo-controlled study in 291 infants aged 12-16 weeks in Tanzania (Lancet 2003; 361: 1853-60). At enrollment, children received their third dose of combined diphtheria-tetanus-pertussis and poliomyelitis expanded vaccines with the first of three daily doses of amodiaquine intermittent preventive treatment (IPTi) or placebo. After 60 days, children receiving amodiaquine had significantly fewer malaria fevers than controls. WHERE NEXT: The increasing concordance of malaria control and vaccination, movement toward co-administration of IPTi with immunisation, and the increase in travellers to malarious areas who receive concurrent vaccinations and chemoprophylaxis warrant further study. PMID- 15110500 TI - Health-care system frailties and public health control of communicable disease on the European Union's new eastern border. AB - In May, 2004, the border of the European Union (EU) will shift eastward such that the new frontier will be made up by Ukraine, Belarus, and a considerably longer Russian border. Here, we discuss three issues: first, the factors that have contributed to the growth of communicable disease in Russia, Ukraine, and Belarus; second, how public health systems have responded to these challenges; and third, the implications for the EU as a whole. Since the break-up of the Soviet Union, Russia, Ukraine, and Belarus have witnessed substantial political, social, and economic changes. These events have been reflected in changes in the epidemiology of communicable diseases, including tuberculosis and HIV (ie, HIV 1). Moreover, public health systems, rooted in Soviet traditions, are struggling to respond effectively to the challenges of resurgent infectious diseases, such as tuberculosis, and newly emergent challenges such as HIV. The changing patterns of communicable diseases east of the EU's new border has implications for how the EU aids the strengthening of public health systems east of the new frontier. Transborder spread of communicable diseases also challenges communicable disease control systems within the EU. Concerted action is needed by member states and the EU, building on models of cooperation between institutions that have been successful in areas beyond health, if public health systems are to meet the emerging challenges to communicable disease control. PMID- 15110501 TI - Scar of Japan's leprosy isolation policy in Korea. PMID- 15110502 TI - The Global Fund and effective malaria treatment. PMID- 15110503 TI - Science and ideology. PMID- 15110504 TI - Diverticular disease of the colon. PMID- 15110505 TI - Diverticular disease of the colon. PMID- 15110506 TI - Migraine. PMID- 15110507 TI - Migraine. PMID- 15110510 TI - HIV-1 and hepatitis B virus: time for a re-think? PMID- 15110509 TI - Hospital stay in patients with non-ST-elevation acute coronary syndromes. PMID- 15110511 TI - Is inseki-jisatsu, responsibility-driven suicide, culture-bound? PMID- 15110512 TI - Where has vancomycin-heterogeneously resistant Staphylococcus aureus gone? PMID- 15110513 TI - Laicite in hospitals. PMID- 15110514 TI - High mortality in displaced populations of northern Uganda. PMID- 15110515 TI - Photographic faux pas. PMID- 15110517 TI - Medicine as a black box. PMID- 15110516 TI - Perri Klass. PMID- 15110519 TI - Propagation, infection, and neutralization of authentic HPV16 virus. AB - Despite the prevalence of HPV16 in invasive cervical cancers, an in vitro system capable of producing infectious HPV16 is lacking. The organotypic (raft) culture system has allowed for the study of the entire differentiation-dependent life cycle of human papillomaviruses (HPVs). However, the use of this system with the prototype HPV16-containing cell line, W12, has failed to yield infectious virus. Our laboratory has introduced clinically derived HPV16(114/B) genomic DNA into primary keratinocytes, where it subsequently recircularized and maintained episomally at 50-100 copies per cell. Virion morphogenesis occurred after epithelial stratification and differentiation in raft culture. HPV16 virions were isolated that were able to infect keratinocytes in vitro. Infection was neutralized by monoclonal antibodies raised against HPV16 but not by monoclonal antibodies known to neutralize other HPV types. PMID- 15110520 TI - Morphological changes contribute to apoptotic cell death and are affected by caspase-3 and caspase-6 inhibitors during red sea bream iridovirus permissive replication. AB - Red sea bream iridovirus (RSIV) of the Iridoviridae family is a causative agent of lethal infections in many cultured marine fish species in southwestern Japan. RSIV-induced apoptosis was divided as follows: (1). cell shrinkage and rounding at the early apoptotic stage, (2). cell enlargement at the middle apoptotic stage, (3). formation of apoptotic body-like vesicles at the late apoptotic stage and phagocytosis by neighboring cells, and (4). loss of membrane integrity in apoptotic body-like vesicles without phagocytosis by neighboring cells. By affinity labeling, RSIV-induced apoptosis included caspase-dependent apoptosis. RSIV infection caused cell rounding but not cell enlargement or formation of apoptotic body-like vesicles and further restricted part of the structural protein synthesis in the presence of caspase-3 and -6 inhibitors. These findings showed the involvement of caspase-3 and -6 in the morphological changes at the middle and late apoptotic stages and viral protein synthesis in the late stage of RSIV infection. PMID- 15110521 TI - Does Toll-like receptor 3 play a biological role in virus infections? AB - The Toll-like receptor (TLR) family functions to recognize conserved microbial and viral structures with the purpose of activating signal pathways to instigate immune responses against infections by these organisms. For example, in vitro studies reveal that the TLR3 ligand is a double-stranded RNA (dsRNA), a product of viral infections. From this observation, it has been proposed that TLR3 is likely an important first signal for virus infections. We approached this issue by investigating the role of TLR3 in four different infectious viral models (lymphocytic choriomeningitis virus (LCMV), vesicular stomatitis virus (VSV), murine cytomegalovirus (MCMV), and reovirus) and in TLR3 genetically deficient (( /-)) mice. Our results indicate that TLR3 is not universally required for the generation of effective antiviral responses because the absence of TLR3 does not alter either viral pathogenesis or impair host's generation of adaptive antiviral responses to these viruses. PMID- 15110522 TI - Genomic organization and molecular analysis of the inducible prophage EJ-1, a mosaic myovirus from an atypical pneumococcus. AB - We report the complete genomic sequence of EJ-1, an inducible prophage isolated from an atypical Streptococcus pneumoniae strain that belongs to the Myoviridae morphology family. The phage and bacterial recombinational sites (attachment sites) have been also determined. The genome of the EJ-1 prophage (42935 bp) is organized in 73 open reading frames (ORFs) and in at least five major clusters. Bioinformatic and N-terminal amino acid sequence analyses enabled the assignment of possible functions to 52 ORFs. The predicted proteins coded for the EJ-1 genome revealed similarities in the lysogeny, DNA replication, regulation, packaging, and head morphogenesis protein clusters with those from several siphoviruses infecting lactic acid bacteria. However, the proteins encoded by genes orf53 to orf64, corresponding to putative tail proteins of the virion, were very similar to those of the defective Bacillus subtilis myovirus PBSX with the notable exception of the gene product of orf56 (the tape measure tail protein) that was similar to proteins from phages infecting Gram-negative bacteria. The first description of the genome of a myovirus infecting a low G + C content Gram positive bacterium, a member of a group embracing important human pathogens and industrial relevant species, will contribute to expand our current knowledge on phage biology and evolution. PMID- 15110523 TI - The portal protein plays essential roles at different steps of the SPP1 DNA packaging process. AB - A large number of viruses use a specialized portal for entry of DNA to the viral capsid and for its polarized exit at the beginning of infection. These families of viruses assemble an icosahedral procapsid containing a portal protein oligomer in one of its 12 vertices. The viral ATPase (terminase) interacts with the portal vertex to form a powerful molecular motor that translocates DNA to the procapsid interior against a steep concentration gradient. The portal protein is an essential component of this DNA packaging machine. Characterization of single amino acid substitutions in the portal protein gp6 of bacteriophage SPP1 that block DNA packaging identified sequential steps in the packaging mechanism that require its action. Gp6 is essential at early steps of DNA packaging and for DNA translocation to the capsid interior, it affects the efficiency of DNA packaging, it is a central component of the headful sensor that determines the size of the packaged DNA molecule, and is essential for closure of the portal pore by the head completion proteins to prevent exit of the DNA encapsidated. Functional regions of gp6 necessary at each step are identified within its primary structure. The similarity between the architecture of portal oligomers and between the DNA packaging strategies of viruses using portals strongly suggests that the portal protein plays the same roles in a large number of viruses. PMID- 15110524 TI - Immunogenicity and T cell recognition in swine of foot-and-mouth disease virus polymerase 3D. AB - Immunization of domestic pigs with a vaccinia virus (VV) recombinant expressing foot-and-mouth disease virus (FMDV) 3D protein conferred partial protection against challenge with infectious virus. The severity reduction of the clinical symptoms developed by the challenged animals occurred in the absence of significant levels of anti-3D circulating antibodies. This observation suggested that the partial protection observed was mediated by the induction of a 3D specific cellular immune response. To gain information on the T cell recognition of FMDV 3D protein, we conducted in vitro proliferative assays using lymphocytes from outbred pigs experimentally infected with FMDV and 90 overlapping peptides spanning the complete 3D sequence. The use of pools of two to three peptides allowed the identification of T cell epitopes that were efficiently recognized by lymphocytes from at least four of the five animals analyzed. This recognition was heterotypic because anti-peptide responses increased upon reinfection of animals with a FMDV isolate from a different serotype. The results obtained with individual peptides confirmed the antigenicity observed with peptide pools. Detection of cytokine mRNAs by RT-PCR in lymphocytes stimulated in vitro by individual 3D peptides revealed that IFN-gamma mRNA was the most consistently induced, suggesting that the activated T cells belong to the Th 1 subset. These results indicate that 3D protein contains epitopes that can be efficiently recognized by porcine T lymphocytes from different infected animals, both upon primary and secondary (heterotypic) FMDV infection. These epitopes can extend the repertoire of viral T cell epitopes to be included in subunit and synthetic FMD vaccines. PMID- 15110525 TI - Reduced incorporation of the influenza B virus BM2 protein in virus particles decreases infectivity. AB - BM2 is the fourth integral membrane protein encoded by the influenza B virus genome. It is synthesized late in infection and transported to the plasma membrane from where it is subsequently incorporated into progeny virus particles. It has recently been reported that BM2 has ion channel activity and may be the functional homologue of the influenza A virus M2 protein acting as an ion channel involved in viral entry. Using a reverse genetic approach it was not possible to recover virus which lacked BM2. A recombinant influenza B virus was generated in which the BM2 AUG initiation codon was mutated to GUG. This decreased the efficiency of translation of BM2 protein such that progeny virions contained only 1/8 the amount of BM2 seen in wild-type virus. The reduction in BM2 incorporation resulted in a reduction in infectivity although there was no concomitant decrease in the numbers of virions released from the infected cells. These data imply that the incorporation of sufficient BM2 protein into influenza B virions is required for infectivity of the virus particles. PMID- 15110526 TI - Comparative usage of herpesvirus entry mediator A and nectin-1 by laboratory strains and clinical isolates of herpes simplex virus. AB - The herpesvirus entry mediator A (HVEM/HveA) and nectin-1 (HveC/CD111) are two major receptors for herpes simplex virus (HSV). Although structurally unrelated, both receptors can independently mediate entry of wild-type (wt) HSV-1 and HSV-2 by interacting with the viral envelope glycoprotein D (gD). Laboratory strains with defined mutations in gD (e.g. rid1) do not use HVEM but use nectin-2 (HveB/CD112) for entry. The relative usage of HVEM and nectin-1 during HSV infection in vivo is not known. In the absence of a defined in vivo model, we used in vitro approaches to address this question. First, we screened HSV clinical isolates from various origins for receptor tropism and found that all used both HVEM and nectin-1. Second, we determined the numbers of surface receptors on various susceptible and resistant cell lines as well as on primary fibroblasts derived from an individual with cleft lip/palate ectodermal dysplasia (CLPED1). Although CLPED1 cells can only express a defective form of nectin-1, they allowed entry of wild type and mutant HSV strains by usage of either HVEM or nectin-2. Finally, we compared the ability of HVEM and nectin-1 to mediate entry when expressed at varying cell surface densities. Both receptors showed a direct relationship between the number of receptors and HSV susceptibility. Direct comparison of receptors suggests that nectin-1 is more efficient at promoting entry than HVEM. Overall, our data suggest that both receptors play a role during HSV infection in vivo and that both are highly efficient even at low levels of expression. PMID- 15110527 TI - Influenza-induced tachypnea is prevented in immune cotton rats, but cannot be treated with an anti-inflammatory steroid or a neuraminidase inhibitor. AB - Influenza viruses are one of the leading causes of morbidity and mortality during winter months. Increased respiratory rate (tachypnea) is a sign of increasing lower respiratory disease during influenza infection and is frequently observed in hospitalized patients. We investigated this clinical sign in influenza virus infected cotton rats (Sigmodon hispidus) and the efficacy of antiviral and anti inflammatory therapy in reducing symptomatic disease. Cotton rats infected intranasally with A/Wuhan/359/95 (H3N2) had increased respiratory rates from 1 to 4 days postinfection that correlated with the dose of virus used to inoculate the animal but not the amount of virus recovered from the lung. In addition, evaluation of sequential lung tissue pathology revealed that extensive epithelial cell destruction of small airways correlated with tachypnea. Increased respiratory rate was not observed in immune animals, supporting results that demonstrated a requirement for exposure to, and infection by, large amounts of live virus for induction of tachypnea. A variety of therapeutic approaches proved ineffective in reducing tachypnea, including anti-inflammatory therapy with systemic triamcinolone acetonide, bronchodilatory therapy with levalbuterol, or antiviral therapy with zanamivir. These results, together with the pathologic observations, suggest that early disruption of the lower respiratory tract epithelium is a major component of the pathophysiology of influenza infection. Therapeutic approaches need to be tailored to clear airway obstruction and restore an intact epithelium. PMID- 15110528 TI - Specific antibodies modulate the interactions of adenovirus type 5 with dendritic cells. AB - Adenovirus type 5 (Ad5) is able to induce an efficient CD8+ T lymphocyte (CTL) response against a transgene product, a property thought to be linked to its ability to transduce dendritic cells (DCs). Little, however, is known about the capacity of Ad5 to interact with DCs in the presence of specific antibodies, although most people test positive for antibodies directed against Ad5. In the present study, we found that in the presence of Ad5 antibodies, a large fraction of Ad5 binds very efficiently to DCs, and that this binding is FcgammaRII/FcgammaRIII dependent. Nevertheless, in the presence of high levels of antibodies against the whole virion, Ad5 entry was inhibited. Increased binding led to increased entry in DCs in the presence of fiber-specific antibodies or in the presence of low amounts of a whole antiserum raised against whole virions, showing that the relative concentration of antibodies directed against fiber and penton base plays a major role in entry efficacy. Nevertheless, mice previously immunized with virions or purified fiber developed a lower transgene-specific CD8+ T cell response than naive mice, although their serum appeared to increase virus entry into DCs in vitro. PMID- 15110529 TI - A new model of Hantaan virus persistence in mice: the balance between HTNV infection and CD8(+) T-cell responses. AB - We established a viral persistence model that involves the adoptive transfer of spleen cells from immunocompetent mice (H-2(d)) into Hantaan virus (HTNV) infected severe combined immunodeficient (SCID, H-2(d)) mice. The infection is maintained despite the presence of neutralizing antibodies, without apparent signs of disease, and there is a correlation between HTNV persistence and the lack of HTNV-specific CD8(+) T cells. In addition, disseminated HTNV infection before the initiation of immune responses appears to be important for virus persistence. The suppression of HTNV-specific CD8(+) T cells in the present model appears to occur at the periphery. The present study also demonstrates that CD8(+) T cells contribute to the clearance of HTNV. Thus, it seems that HTNV specific CD8(+) T cells play a key role in HTNV persistence in mice. This model of viral persistence is useful for studies of immune responses and immunocytotherapy against viral infection. PMID- 15110530 TI - Phospholipid molecular species profiles of tectiviruses infecting Gram-negative and Gram-positive hosts. AB - The phospholipid (PL) molecular species compositions of bacteriophages PRD1 and Bam35 as well as their respective hosts were determined quantitatively using liquid chromatography/electrospray ionization mass-spectrometry (LC-ESI-MS) and backed up by gas-chromatographic/mass-spectrometric (GC-MS) analysis of the total fatty acids (FAs). The results showed that both viruses contain significantly more phosphatidylglycerol (PG) and less phosphatidylethanolamine (PE) than the host membranes. Only modest differences in the molecular species composition of the viruses and their respective hosts were observed, indicating that the virus assembly process is relatively nonselective in respect of the fatty acid (FA) proportion of phospholipids (PL). These data indicate that the PL composition of these two viruses is largely, albeit not exclusively, determined by the availability of phospholipids in the host membrane. PMID- 15110531 TI - Improved protection conferred by vaccination with a recombinant vaccinia virus that incorporates a foreign antigen into the extracellular enveloped virion. AB - Recombinant poxviruses have shown promise as vaccine vectors. We hypothesized that improved cellular immune responses could be developed to a foreign antigen by incorporating it as part of the extracellular enveloped virion (EEV). We therefore constructed a recombinant vaccinia virus that replaced the cytoplasmic domain of the B5R protein with a test antigen, HIV-1 Gag. Mice immunized with the virus expressing Gag fused to B5R had significantly better primary CD4 T-cell responses than recombinant virus expressing HIV-Gag from the TK-locus. The CD8 T cell responses were less different between the two groups. Importantly, although we saw differences in the immune response to the test antigen, the vaccinia virus specific immune responses were similar with both constructs. When groups of vaccinated mice were challenged 30 days later with a recombinant Listeria monocytogenes that expresses HIV-Gag, mice inoculated with the virus that expresses the B5R-Gag fusion protein had lower colony counts of Listeria in the liver and spleen than mice vaccinated with the standard recombinant. Thus, vaccinia virus expressing foreign antigen incorporated into EEV may be a better vaccine strategy than standard recombinant vaccinia virus. PMID- 15110532 TI - Adenovirus serotype 3 utilizes CD80 (B7.1) and CD86 (B7.2) as cellular attachment receptors. AB - Most viruses exploit a variety of host cellular proteins as primary cellular attachment receptors in the context of successful execution of infection. Furthermore, many viral agents have evolved precise mechanisms to subvert host immune recognition to achieve persistence. Herein we present data indicating that adenovirus (Ad) serotype 3 utilizes CD80 (B7.1) and CD86 (B7.2) as cellular attachment receptors. CD80 and CD86 are co-stimulatory molecules that are present on mature dendritic cells and B lymphocytes and are involved in stimulating T lymphocyte activation. To our knowledge, this is one of the first demonstrations of a virus utilizing immunologic accessory molecules as a primary means of cellular entry. This finding suggests a mechanism whereby viral exploitation of these proteins as receptors may achieve both goals of cellular entry and evading the immune system. PMID- 15110533 TI - Dissection of seroreactivity against the tryptophan-rich motif of the feline immunodeficiency virus transmembrane glycoprotein. AB - Immunogenicity of the tryptophan-rich motif (TrpM) in the membrane-proximal ectodomain of the transmembrane (TM) glycoprotein of feline immunodeficiency virus (FIV) was investigated. Peptide 59, a peptide containing the TrpM of the TM of FIV, was covalently coupled to Qbeta phage virus-like particles (Qbeta-59) in the attempt to induce potent anti-TrpM B cell responses in cats. All Qbeta-59 immunized cats, but not cats that received a mixture of uncoupled Qbeta and peptide 59, developed antibodies that reacted with a same epitope in extensive binding and binding competition assays. The epitope recognized was composed of three amino acids, two of which are adjacent. However, Qbeta-59-immune sera failed to recognize whole FIV in all binding and neutralization assays performed. Furthermore, no reactivity against the TrpM was detected by screening sera from FIV-infected cats that had reacted with TM peptides, confirming that this epitope does not seem to be serologically functional in the FIV virion. The data suggest that TrpM may not be a suitable target for antiviral vaccine design. PMID- 15110535 TI - In-line ultrasonic monitoring of semi-solid magnesium die casting process. AB - Ultrasonic velocity and attenuation measurements in AZ91D magnesium (Mg) alloy with dendritic, rosette and globular microstructures were performed at elevated temperatures using a non-contact laser-ultrasonic technique. It was found that the ultrasonic velocity in the globular microstructure and the ultrasonic attenuation in the dendritic microstructure are the highest among the three microstructures. An ultrasonic clad steel buffer rod sensor embedded in the die has been used to monitor the semi-solid die casting process in-line for the AZ91D Mg alloy. This probe monitored the completion of the die filling, the release of the pressure, the opening of the die, part detachment, solidification of the part, the averaged temperature of the die and the part. PMID- 15110534 TI - Rotavirus RRV associates with lipid membrane microdomains during cell entry. AB - Rotavirus cell entry is a multistep process, not completely understood, which requires at least four interactions between the virus and cell surface molecules. In this work, we investigated the role of the sphingolipid- and cholesterol enriched lipid microdomains (rafts) in the entry of rotavirus strain RRV to MA104 cells. We found that ganglioside GM1, integrin subunits alpha2 and beta3, and the heat shock cognate protein 70 (hsc70), all of which have been implicated as rotavirus receptors, are associated with TX-100 and Lubrol WX detergent-resistant membranes (DRMs). Integrin subunits alpha2 and beta3 were found to be particularly enriched in DRMs resistant to lysis by Lubrol WX. When purified RRV particles were incubated with cells at 4 degrees C, about 10% of the total infectious virus was found associated with DRMs, and the DRM-associated virus increased to 37% in Lubrol-resistant membrane domains after 60-min incubation at 37 degrees C. The virus was excluded from DRMs if the cells were treated with methyl-beta-cyclodextrin (MbetaCD). Immunoblot analysis of the viral proteins showed that the virus surface proteins became enriched in DRMs upon incubation at 37 degrees C, being almost exclusively localized in Lubrol-resistant DRMs after 60 min. These data suggest that detergent-resistant membrane domains play an important role in the cell entry of rotaviruses, which could provide a platform to facilitate the efficient interaction of the rotavirus receptors with the virus particle. PMID- 15110536 TI - The use of ultrasonic guided waves and wavelets analysis in pipe inspection. AB - One of the main applications of guided waves is for pipe inspection, since its one-dimensional geometry allows inspecting long distances in a short time. As with most configurations (frequency, thickness) ultrasonic waves with many modes of propagation are generated, recent research focuses mostly on the generation and reception of specific modes, mainly by means of sophisticated arrays of transducers. In this paper this problem is addressed with a different approach, by processing ultrasonic signals with a low signal/noise ratio acquired with a single transducer in an pulse-echo configuration. In order to improve the evaluation of results, frequency bandpass filters and wavelet analysis were tested. Results showed that even when very noisy signals are utilized, signal processing improve the signal/noise (S/N) ratio up to 12 dB approximately and enhance the analysis of the results, thus demonstrating its usefulness. PMID- 15110537 TI - A new technique for the identification of ultrasonic flaw signals using deconvolution. AB - The identification of ultrasonic flaw signals is a difficult task in the angle beam ultrasonic testing of welded joints due to the presence of non-relevant signals from the geometric reflectors such as weld-roots and counter-bores. This paper describes a new approach to identify ultrasonic flaw signals in such a problematic situation. A similarity function is defined as the deconvolution of a target signal by a reference signal. The similarity functions for the same type of flaws/references are symmetric bandlimited impulse-like patterns with larger amplitudes while those for different types of flaws/references are asymmetrical broad patterns with relatively smaller amplitudes. Therefore, ultrasonic signals could be identified by the pattern of the similarity function. In the initial experiments, the proposed technique showed great potential for identifying ultrasonic flaw signals in the inspection of weld joints. PMID- 15110538 TI - Flow acoustics modelling and implications for ultrasonic flow measurement based on the transit-time method. AB - A comparison between three mathematical models frequently used in flow acoustics is presented and discussed with respect to ultrasonic flow-meter performance based on the transit-time method. The flow-meter spoolpiece geometry is assumed to be a cylindrical pipe. Semi-analytical calculations employing the Frobenius power series expansion method are shown for the cases of a constant-, linear-, parabolic-, and cubic-flow profiles although the Frobenius method presented can be applied to any smooth flow profile. It is shown that the so-called deviation of measurement, often used as a measure of the flow-meter accuracy, is strongly dependent on the acoustic mode excited and the flow profile. Furthermore, differences with respect to deviation of measurement results exist among the three mathematical models analyzed. PMID- 15110539 TI - Non-destructive testing of tubes using a time reverse numerical simulation (TRNS) method. AB - A method for the detection of defects in cylindrical structures and the determination of their positions and orientations is presented in this paper. The scattered field, which is generated by the interaction of excited guided waves with a defect, is evaluated with an approach named time reverse numerical simulation method (TRNS). Since the excited waves and the scattered field propagate along the sample, the time-consuming scanning of the whole tube can be eliminated. The scattered displacement field is measured in three dimensions over time with a laser vibrometer at different locations distributed equally around the circumference at a fixed axial coordinate far away from the defect. Instead of analyzing the complicated time signals directly, they are played back in time. If the recorded displacement histories of the scattered field are reversed in time and played back in an identical structure, the waves travel back the same path and interfere to a maximum at their origin. The result is an amplitude increase at the position of the defect where the scattered field was generated. Instead of playing back the recorded time signals in an experiment, this step is replaced by a numerical simulation. Only this enables the visualization and detection of the amplitude increase. As long as the simulation is of high accuracy, the position of the maximum interference corresponds exactly to the location of the defect in the experiment, although no defect is implemented in the simulation. PMID- 15110541 TI - Medicolegal aspects of donor safety evaluation. AB - Current Italian statutes concern the safety of organ donors. The law of April 1, 1999 (no. 91), established the fundamental principles for a new structure for organ transplants in Italy and, therefore, for the quality of transplanted organs. The Ministry of Health decree of February 8, 2002, defined organ conditions that preclude the utilization of an organ and identified conditions for suitability of non-optimal organs for some types of transplants or for some recipients. Guidelines from the National Transplant Center establish risk levels, criteria for absolute exclusion, and standard procedures for the evaluation of donor risk and organ suitability for transplant. This article also examines the levels of responsibility of various professionals involved in the harvesting transplant process. PMID- 15110542 TI - Summary: the quality improvement program in organ donation of the Tuscany region. AB - Since October 2002, after a pilot period, a quality improvement program in organ donation has been underway in Tuscany. This program is based on a database elaborated by the Transplant Coordination Office of the Pisa University, according to the Spanish program of the National Transplant Organization. All encephalic deaths occurring in intensive care units beds during mechanical ventilation are registered in the database. Encephalic deaths were evaluated by local transplant coordinators and an esthesiologists after review of the clinical records. The data are sent every month to the Central Unit located in the Santa Chiara Hospital of Pisa. Every 3 and 12 months, we calculate the indices to evaluate organ donation activity in every hospital. The preliminary results show that: (1) the program is a useful tool to evaluate the organ donation process; (2) the experience is limited, but has shown the potential of the program to increase organ donation activity in Tuscany. PMID- 15110543 TI - Donor and organ suitability: risk due to donor social behavior. AB - For many patients experiencing organ failure, transplantation may be the only opportunity to improve their quality of life and to extend life expectancy. Organ shortage is an important problem for Italian and international procurement organizations. A review of donor and organ suitability is necessary to expand the donor pool. The suitability of donors at risk for HIV infection, as well as other marginal individuals, must be carefully evaluated before rejecting them. For donors with a social history considered to be at risk for HIV infection, laboratory testing is necessary to establish the actual HIV infection risk. It is important to consider every organ available for transplantation, even those considered to be marginal or coming from marginal donors. The only condition that we must always consider is that the organs be of acceptable quality and, above all, not expose patients to unacceptable risks. PMID- 15110544 TI - Opinions on renal transplantation and organ donation in high school students in two large northern (Torino) and southern (Napoli) Italian cities. AB - Organ shortage for transplantation has focused attention on educational interventions. Italy is a nonhomogenous country whose cultural and economic differences are reflected in the health-care system: dialysis is mainly public in the north versus private in the south; and transplantation rates display a wide range from 3.4 to 37.8 per million people in 2002. The aim of the present study was to analyze the opinions of population of high school students (last two years) in two large cities: northern (Torino) and southern (Napoli) Italy, as a knowledge base for a randomized controlled trial on the efficacy of educational interventions on renal replacement therapy and organ donation, targeted to high school students. METHODS: This preliminary study included eight public high schools that completed a first and anonymous semistructured questionnaire. Five hundred and eighty nine questionnaires were retrieved in Torino and 539 in Napoli. In both cities most students answered that they would give a kidney to a brother, sister, or partner needing dialysis (Torino: yes 80.6%; no 2.2%, uncertain-blank 17.2%; Napoli: yes 86.1%, no 1.1%; uncertain-blank 12.8%). Only 36.3% of the students in Torino and 37.7% in Napoli answered that they would consent to organ donation, if they had to choose for a strict relative with brain death. Opposition was 28% in Torino and 23.7% in Napoli; 35.7% in Torino and 38.6% in Napoli were blank-uncertain. These data underline the need for detailed information on the opinions of the overall population as basis for tailored educational campaigns. PMID- 15110545 TI - Efficacy of an educational program on dialysis, renal transplantation, and organ donation on the opinions of high school students: a randomized controlled trial. AB - AIM: Organ shortage is a rate-limiting factor for transplantation. The aim of this study was to evaluate the impact of an educational program targeted to high school students on opinions concerning organ donation. METHODS: Sixteen public high schools in Torino, Italy, were randomized (2001 to 2002) as interventions (n = 8) or controls (n = 8). Intervention was composed of first questionnaire, first lesson (one to two classes; 2 hours, by a trained nephrology fellow); second lesson (all classes together; coordinated by a nephrologist, with patients and trainees); second questionnaire. Control included questionnaires. Statistical analysis compared the opinions in the questionnaires after stratification for age, sex, and type of school. RESULTS: Fourteen schools completed the program (seven interventions: 937 first and 808 second questionnaires; controls: 739 and 659). Television (82.5%) and newspapers (43.2%) were the main sources of information; knowledge on renal transplantation (grafts feasible per patient, average duration) was low; only 12.2% of the students gave correct answers. The opinions on living donation were highly positive (76.8%) with no difference in control, intervention schools, first and second questionnaires, according to sex, age, or type of school. The opinions on cadaveric transplantation were affected by the educational intervention with a drop in negative answers (from 33.7% to 16%), with an increase in positive (from 31.5% to 42.9%) and in uncertain ones (from 34.8% to 41.1%) among the intervention schools; 98% of the students appreciated the program. CONCLUSION: The positive effect on student opinions suggests the need to develop educational approaches as a part of our routine clinical work. PMID- 15110546 TI - Preliminary results of an expert-opinion elicitation process to prioritize an informative system funded by Italian Ministry of Health for cadaveric donor management, organ allocation, and transplantation activity. AB - Expert-opinion elicitation (EOE) is a heuristic process for gathering evidence and data or answering questions on issues/problems of concern. The Delphi method (DM) is the most frequent technique used to obtain structured elicitation of expert opinions. It has been increasingly applied in medicine to produce guidelines and to evaluate the appropriateness of diagnostic procedures. In this study, a DM has been used as structured EOE process to assess the appropriateness and clinical priority of a data set-based informative system in the context of the Liguria-Trento Transplant Network (LTTN) Project, funded by Italian Ministry of Health. The original data set was obtained by using an interdisciplinary pool of regional experts (n = 60). This data set held 1506 items stratified in 21 categories at various surgical phases (preoperative, intraoperative, and postoperative) and transplantation types (liver, kidney, and kidney/pancreas) in adult and pediatric recipients. Some categories included cadaveric donor management, organ allocation, and acute liver failure. In the second DM round, the data set was subjected to a panel of extraregional, independent experts (n = 9) to assess scores ranging from 1 to 9 on each item, based on increasing appropriateness/priority, according to RAND/UCLA Appropriateness Method. The overall agreement between experts was 95.88%, whereas disagreement and uncertainty were 0.13% and 3.98%, respectively. A major uncertainty occurred for the data set concerning the multiorgan cadaveric donor, for liver transplantation, and for kidney transplantation in adult recipients. The use of a structured EOE process may represent an effective strategy to define the appropriateness and prioritization criteria of a large data set in the field of solid organ transplantation. PMID- 15110547 TI - Regional procurement team for abdominal organs. AB - BACKGROUND: The graft shortages make multiorgan procurements mandatory. We describe the results of a regional procurement team policy that has been employed over a 5-year period. METHODS: Three hundred forty-three multiorgan procurements were performed by a regional team using an en bloc harvesting method. RESULTS: Among 1374 grafts procured, none was discarded because of iatrogenic injuries. In three instances the liver, the pancreas, and the small bowel were procured simultaneously and transplanted to different recipients. In 42 instances the liver was not allocated to our center. Forty liver teams (95%) from 11 institutions agreed to allow the regional procurement team to run the donor procedure. CONCLUSIONS: Our experience confirms that a regional team can successfully manage most multiorgan procurements including complex donor procedures, such as simultaneous procurement of liver, pancreas, and intestine from the same donor for transplantation to different recipients. PMID- 15110548 TI - Outcome of isolated small bowel and pancreas transplants retrieved from multiorgan donor: the in vivo technique. AB - Even when considering the possibility of organ rejection and the complications of immunosuppression, the risks associated with total parenteral nutrition therapy are life-threatening. Therefore, for patients with end-stage bowel disease small bowel transplantation (SBTx) is the only therapeutic option. The preferred method to procure these organs is debated, especially when, graft retrieval is associated with concurrent abdominal organ procurement of the pancreas, which shares part of the vascular inflow and outflow with the small bowel. While many surgeons procure the graft using the en bloc method, dissecting tissue at the back table, our preference is to use an in vivo technique, which results in shorter cold ischemia times and less bleeding during reperfusion of the pancreas/small bowel as well as decreased ascites production during the postoperative period and less edema and capsular bleeding of the pancreatic grafts. This article presents an analysis of 19 multiorgan cadaveric procurements using the in vivo technique with a focus on the quality of pancreas/small bowel postreperfusion properties during the first 5 to 6 postoperative months. PMID- 15110549 TI - Organizzazione Centro-Sud Trapianti: outcome analysis 1999 to 2002. AB - BACKGROUND: The Organizzazione Centro-Sud Trapianti (OCST), which was created in 1998, is organized into eight regional areas, each referring to a local Regional Transplant Coordinating Center. Organs are primarily allocated to meet the demands of transplant centers in each regional area. Urgencies, pediatric grafts, and paybacks are managed by an Interregional Transplant Coordinating Center. The aim of the current work is to report on the impact of introduction of OCST on organ donation and transplant activity over the period from 1999 to 2002. MATERIALS AND METHODS: A retrospective analysis of donor and transplant data charts over the period from 1999 to 2002 focused on outcome analysis based on donor epidemiological data, cause of death, reasons for discards and grafts performed at OCST local transplant centers. RESULTS: From 1999 to 2002, we observed a remarkable increase in organ donation from 8.8 to 22.5 donors per million people. Donor epidemiology showed an increase in median age and stroke incidence rates and a decrease in trauma cases. The nonharvested donor rate rose steadily over the study period, plateauing at 58%, which was compensated for by a threefold increase in donation. Family oppositions ranged as high as 35.5% on average, despite public efforts to support donation. Transplant activity rose by 76%. CONCLUSIONS: The institution of OCST and the efforts from central and regional authorities have yielded a significant increase in organ donation and transplant activity rates over the period from 1999 to 2002. Major areas of concern are the high opposition rate and the decreasing quality of harvested grafts. Long-term analysis is underway to assess the impact of OCST on the quality of transplants performed in the catchment area. PMID- 15110550 TI - First results of Liguria-Trento transplant network project: a model for a macroregional network and real-time registry in Italy. AB - In Italy, health-care telematic is funded and supported at the level of national government or regional institutions. In 1999, the Italian Ministry of Health started to fund the Liguria-Trento Transplant Network (LTTN) project, a health research project with the aim to build an informative system for donor management and transplantation activity in a macroregional area. At the time of LTTN project proposal, no published Transplant Network Informative System fulfilled Italian rules on telematic management of electronic documentation concerning transplantation activity. Partnership of LTTN project were two Regional Transplant Coordinating Centres, Nord Italia Transplant Interregional Coordinating Centre and the Italian Institute of Health/National Transplant Coordinating Centre. Project Total Quality Management methods were adopted. Technological and case analysis followed ANSI-HL7, CEN-TC251, and Object-Oriented Software Engineering standards. A low-tech prototype powered by a web access relational database is running on a transplant network including web-based clients located in 17 intensive care units, in Nord Italia Transplant Interregional Coordinating Centre, and at the Italian Institute of Health/National Transplant Coordinating Centre. LTTN registry includes pretransplant, surgical, and posttransplant phases regarding liver, kidney, pancreas, and kidney-pancreas transplantation in adult and pediatric recipients. Clinical specifications were prioritized in agreement with the RAND/UCLA appropriateness method. Further implementation will include formal rules for data access and output release, fault tolerance, and a continuous registry evolution plan. PMID- 15110551 TI - How many organs should one patient receive? The ethics of transplantation in the medical school. AB - BACKGROUND: Interest in the humanities in the medical school is growing; while several medical schools, mainly of Anglo-Saxon background, have developed dedicated courses, the experience in Italy is limited. METHODS: Since the academic year 2000 to 2001, a discussion of ethical problems was implemented in the nephrology course (fourth year of the Medical School of Torino, Italy; overall 6 years). In 2002 to 2003, a case entitled "Retransplantation of Multiple Organs (Prog Transplant 2002)" was discussed in 2 hours of small-group tutorial teaching: a boy received a renal graft at age 5, failed at age 7 due to recurrent glomerulonephritis, required a heart-kidney graft at age 11, and a second heart kidney graft at 17. Student opinions were gathered by anonymous semistructured questionnaires at the beginning of the lessons as a basis for discussion. RESULTS: Following the lessons all students returned the questionnaires (n = 104). In the absence of competition for allocation, retransplantation was approved by 76.2%, unacceptable for 1% (22.9% uncertain-blank). With a waiting list of 10 patients, the opinions changed: 32.4% approved transplantation, 6.7% didn't approve it, 60.9% were uncertain. A theoretical categorization into deontological or utilitaristic approaches favored the first (41.9% vs 26.7%), with a high prevalence of blank-uncertain (31.5%); 21.9% of the students would change their opinion was that study head of the Transplant Department. CONCLUSION: Ethical aspects of the medical profession have been discussed with interest by medical school students; the high prevalence of uncertain answers and requests to develop specific tools underline the importance of this educational approach. PMID- 15110552 TI - Kidney vending: opinions of the medical school students on this controversial issue. AB - BACKGROUND: In this era of globalization, in which different cultural and economic barriers are progressively abated, in the context of the development of rapid information networks such as the Internet, physicians are increasingly challenged by clinical and ethical questions. Kidney vending, banned in some countries, legal or tolerated in others, may be the prototype of the ethical aspects of health-care globalization. METHODS: To test the interest and the opinions of medical school students, a simulated case was proposed to students attending a seminar within the nephrology course fourth year of the Medical School of Torino, san Luigi): an Italian patient comes to the nephrologist's office asking for advice on the possibility to legally buy a kidney in a foreign country. The 43 students attending the lesson answered a semistructured questionnaire (15 boys, 28 girls, of median age 23 years). Attendance was within the usual standards (50 students inscribed per year). From the clinical point of view, 11.6% were favorable to kidney vending, 51.2% were contrary, 37.2% were uncertain. From the ethical point of view, no student was pro, 81.4% were contrary, and 18.6% were uncertain. The open comments underline the importance of patient self-determination and of informed consent. Similar opinions were recorded in a nonstructured question: "What should physician's attitude be, in the face of a choice he/she doesn't share?" CONCLUSION: Students' uncertainties and doubts underline the need to discuss ethical scenarios in the clinical teachings of the medical school. PMID- 15110553 TI - To give or to receive? Opinions of teenagers on kidney donation. AB - BACKGROUND: The attitude toward living donation varies widely in the world, for economic and cultural reasons. In Italy, as in other Mediterranean settings, the role of living kidney donation is minor. AIM: To analyze the reasons for this attitude, we gathered data in a general population sample of high school students in a large northern Italian industrial city (Torino, about 900,000 inhabitants). METHODS: Semistructured questionnaires (n = 1676), gathered in 2001 to 2002 in 14 high schools, in the context of an educational program on dialysis, renal transplantation, and organ donation, were analyzed presumably reflecting opinions gathered before the educational intervention. RESULTS: Most students, in the case of a close relative or partner needing dialysis, answer that they would donate a kidney (yes: 78.2%, no: 2.9%, uncertain-blank: 18.9%); receiving a living donor kidney is felt as disturbing: only 57.5% of the students would accept it (no: 5.9%, uncertain-blank: 36.6%), mainly because of fear of long-term problems for the donor. Donation from an older to a younger person is seen more positively than vice versa. CONCLUSION: In our settings, the attitude of the teenagers on living donation is positive; however, while "giving" is positively seen, the presence of unresolved fears is witnessed by the lower acceptance of the idea of "taking." These data suggest to focus on the risks of kidney donation in educational campaigns and in patient-physician information. The positive attitude shared by the teenagers supports the working hypothesis that lack of information is one of the determinants of the low living donor transplantation rate in our area. PMID- 15110554 TI - Preliminary report on impact of pretransplant dialysis on early graft function: peritoneal versus hemodialysis. AB - Delayed graft function and acute renal failure after kidney transplant negatively influence graft outcome. It has been reported that pretransplantation peritoneal dialysis (PD) instead of hemodialysis (HD) correlated with better short-term graft outcome in adult kidney recipients. In this study the impact of PD versus HD was evaluated among pediatric kidney recipients. This study suggested that different forms of dialysis pretransplantation did not affect early graft function among pediatric kidney recipients. PMID- 15110555 TI - Preparation of candidates for renal transplantation: cost analysis. AB - While the costs of renal transplantation are lower than those of dialysis, little is known about the costs of managing the waiting list. We performed a cost analysis of admission and clinical management of a waiting list for renal and pancreas-kidney transplantation. Admission to the waiting list included (1) renal graft from cadaver: minimum cost Euros () 1784.56 for men < 55 years, maximum 2127.85 for women >/= 55 years; (2) pancreas-kidney transplantation: minimum 2475.50 for men, maximum 2540.10 for women >/= 35 years. Check of suitability state on waiting list after 2 and 5 years: minimum 1400.15 for men >/= 55 years (check every 2 years), maximum 1467.07 for women >/=40, <55 years (every 5 years). The differences are related to the imaging techniques: from 43.90 (Doppler ultrasonography) to 283.28 (coronary angiography). Maintenance of the waiting list: minimum cost 1885.21 in the first year and 3187.02 in the (fifth year) for men < 55 years; maximum 2228.50 (first year) and 5116.70 (fifth year) for women >/= 55 years. These results show different costs for recipients on the basis of sex and age ranges, due to the different requirements for imaging tests such as cardiac scintiscan at age >/= 55 years) and economic charges that increase with age. Reduced waiting times allow lowered total costs. This evaluation allowed us to calculate for our region (Piemonte, Northern Italy), the management costs of the patients presently on our waiting list (369 patients at December 31, 2002) from preparation to transplantation as 959,179.18. PMID- 15110556 TI - Echocardiographic evaluation in type 1 diabetic patients on waiting list for isolated pancreas or kidney-pancreas transplantation. AB - Type 1 diabetic patients may display abnormalities of left ventricular geometry and systolic and diastolic function. Patients on the waiting list for solitary pancreas or kidney-pancreas transplantation were evaluated by Doppler echocardiography to assess left ventricular geometry and systolic and diastolic function, and correlate these parameters with clinical characteristics. We evaluated 78 patients including 45 men with an overall mean age of 39.5 +/- 7.2 years and a disease duration of 24 +/- 9.8 years. Among these 78 patients, 13 showed isolated retinopathy, 9 isolated arterial hypertension, 45 concomitant retinopathy and hypertension and overt nephropathy, while 11 were free of complications. The results of our study showed an increased left ventricular mass and abnormal diastolic function among patients with simultaneous target organ complications and with hypertension, as has been reported in many previous studies. In contrast study patients with no complications showed normal left ventricular structure and function. This finding conflicts with data from several reports in the medical literature in which diastolic impairment was present in type 1 diabetic patients at an early stage of disease and with no evident microvascular and macrovascular complications. PMID- 15110557 TI - Laparoscopic living donor nephrectomy in Italy: a national profile. AB - BACKGROUND: There are no data concerning the national experience with laparoscopic live donor nephrectomy (LLDN) in Italy. A survey was therefore conducted in May 2003 to establish current practice patterns and to describe the outcome of this procedure. METHODS: A self-administered questionnaire was mailed to the 37 Italian kidney transplant centers. Items covered each center's attitude toward LLDN, number of cases performed, and the outcome of donors and recipients. RESULTS: The return rate was 100%. The surveyed centers performed 4818 kidney transplants between January 2000 and May 2003, including 401 (8.3%) from living donors of whom 113 (28.2%) used grafts retrieved by laparoscopy. Despite an absolute increase in the number of living donors, the occurrence of a similar trend in cadaveric donation did not significantly change the overall living donor rate. Sixty-eight percent of LLDNs were done at only two centers. There was no mortality or graft loss and only a minor morbidity related to LLDN. Italian transplant surgeons showed a positive attitude toward LLDN; only a few of those not performing it had no plans to begin an LLDN program. CONCLUSIONS: Three years after the first national case, LLDN had not yet change the living donor rate, although an increasing number of donor nephrectomies were now performed by laparoscopy. Overall the results with the new technique are encouraging, although the pattern of diffusion of LLDN between different areas is heterogeneous and will demand continuous efforts on training programs in laparoscopic techniques for transplant surgeons. PMID- 15110558 TI - Perioperative anesthetic management for laparoscopic kidney donation. AB - OBJECTIVES: To report a single-center experience about the perioperative and anesthetic management of laparoscopic living kidney donation. PATIENTS AND INTERVENTIONS: Subjects undergoing laparoscopic (n = 39) (performed between April 2000 and August 2002) and traditional "open" kidney donation (n = 27) received a standard balanced anesthetic technique. However to counterbalance the reported abdominal insufflation-related kidney dysfunction, laparoscopic donors were administered an extra intravascular volume loading with colloid and crystalloid starting on the night before surgery. RESULTS: Laparoscopic donors underwent longer procedures with lower estimated blood losses (P =.0001), were intraoperatively administered higher amounts of intravenous fluids (P <.01), showed less postoperative analgesic requirement (P <.0001), shorter intensive care unit and overall hospitalization (P <.001), quicker resumption of solid oral intake (P <.01), and full return to work (P <.001) with no difference in the postoperative complication rate. Diuresis resumed intraoperatively in all recipients and early graft function did not differ between the two groups, although the serum creatinine declined earlier, but not significantly, in those receiving kidneys procured by the traditional method. No difference was seen in graft rejection rates. DISCUSSION AND CONCLUSIONS: Laparoscopic kidney donation does not require a particularly complex or expensive anesthetic management or approach; as it has been suggested that intra-abdominal hypertension coming from CO(2) insufflation inside the donor's peritoneal cavity may threaten graft function, during laparoscopic kidney donation it is advisable to adopt a strategy for "renal protection." Thus, when a laparoscopic kidney donation is performed at our center, a multidisciplinary approach is commonly adopted based on three key points: perioperative positive volemic balance in donors; intraoperative urinary output of at least 100 mL/h; inflation with an abdominal pressure not exceeding 12 mm Hg. PMID- 15110559 TI - Survival in kidney transplantation from living donors: a single-center experience. AB - BACKGROUND: Transplantation from living donors, in Italy, is still not accepted, in particular those from unrelated donors. The aim of this paper was to present the experience of one transplant center. MATERIALS AND METHODS: Since 1982, 608 transplants were performed from living donors using cyclosporine as the main component of immunosuppressive therapy. Among those, 402 transplants were from related living donors (338 one haplotype pairs and 25 zero haplotypes pairs) and 206 from unrelated living donors (171 spouses and 35 emotionally related subjects). RESULTS: Graft survival at 1, 5, and 10 years showed no statistically meaningful difference between the two groups. A group of 19 transplants performed in predialytic phase patients was compared with a contemporaneous group of 167 transplants performed in patients who were already receiving dialysis. These two groups did not show any statistically meaningful difference in graft survival at 1, 5, or 10 years. DISCUSSION AND CONCLUSIONS: We think that transplants from living donors, whether related or unrelated, must always be proposed as a therapeutic option for end-stage renal disease patients, since they show an higher graft survival than that from cadaveric donors, independent of the compatibility between donor and recipient and independent of the degree of relationship of the pair. Transplantation from living donors definitely is a complementary, not substitutive, program to that from cadaveric donors, which should always be encouraged with awareness campaigns among the population and targeted programs for healthy personnel. PMID- 15110560 TI - Living donor transplant: wider selection criteria. AB - The availability of cadaveric donor organs is insufficient for actual needs. The organ demand increases by 20% per year. Living donor transplant (LDT) may be a valid therapeutical alternative provided one uses proper criteria. LDT provides many advantages, like improved patient and organ survival, short waiting time, and the possibility to carefully plan the procedure. Potential risks include perioperative mortality and renal dysfunction in the kidney donor. At present, kidney LDTs in Italy represent 8% of the total, with an organ survival rate of 97% after 1 year (vs 93% for cadaveric transplants) and donors mortality rate of almost null. Most LDTs are performed from kinsmen. Presently, law no. 458, 26 June 1967, is in force in Italy for kidney LDT and law no. 453, 16 December 1999, for liver LDT. The foundations of LDT are, of course, the recipient's condition, the donor's motivation, and the altruism of the donation. It is desirable that in the future an increasing number of LDT be performed, supported by a careful, widespread health education regarding organ donation from living subjects and by the possibility to obtain insurance for the donor, which has been considered but never provided by actual laws. PMID- 15110561 TI - Living donor kidney transplant in Italy: is the underutilization justified? AB - Living donor transplants (LDtx) represent an underutilized resource in Italy. It is, however, a therapeutic option that deserves greater consideration not only due to the increasing gap between the number of uremic patients on waiting lists (6956) and the number (1464) of cadaveric transplants (CADtx), as evidenced in 2002, but also due to the advantages of LDtx over CADtx. The superiority of LDtx include better graft survival, independent of the donor/recipient relationship, less need for dialytic treatment with preemptive transplants and reduced immunogenicity of the graft due to the brain death-related "cytokine storm." Moreover, some emerging procedures namely laparoscopic nephrectomy instead of open surgery and spiral CT instead of renal angiography namely, reduce the physical and socioeconomic burden of the donor. In the light of these considerations, LDtx should be reconsidered in the Italian scenario of kidney transplantation. PMID- 15110562 TI - Living kidney transplantation: a starting experience. AB - INTRODUCTION: We present our initial experience with living kidney transplantation. PATIENTS AND METHODS: From January 2001 to December 2002, we performed 27 living kidney transplants using immunosuppression with induction basiliximab, cyclosporine (n = 10 patients), or tacrolimus (n = 17), mycophenolate mofetil, and steroids. RESULTS: Nineteen (70.3%) donors were women and 8 (29.7%) were men of mean age 50.6 years. Four donors were over 65 years of age at the time of living donation. Donor morbidity was 5.5%: namely, one wound infection and one asymptomatic acute pancreatitis. There were no differences between the preoperative and the postoperative mean serum creatinines and systolic blood pressure values. All living donors are in good health with a mean serum creatinine of 0.80 mg/dL at a mean follow-up of 15.2 months. Nineteen (70.3%) recipients were men and 8 (29.7%) were women of mean age 36 years. Acute rejection occurred in 6 (22.2%) recipients. It was more common among spousal donors and among cyclosporine-treated recipients. Patient and graft survivals at a mean follow-up of 15.2 months was 100%. CONCLUSIONS: Our early results showed that accurate selection and preoperative management of potential living donors lead to excellent results in kidney transplantation. The health of the living donors was not impaired by the donation. The rate of early postoperative complications was low. Living donor kidney transplantation, in our geographical area with a low-rate of cadaveric donor transplants, is an alternative to expand the donor pool, which offers better results in term of patient and graft survival. PMID- 15110563 TI - Rare phenotype and transplantability in cadaveric kidney transplant. AB - The waiting list for kidney transplants in the Apulia region contains recipients (about 30%) who were never selected for transplant due to the rare HLA antigen phenotypes and homozygosis. Therefore, an algorithm was selected to equilibrate the chance for patients to be selected despite a rare phenotype. We calculated for each patient, the sum (%) of the A, B, DR antigen frequency (total phenotypic frequency; TPF). All the potential recipients were grouped into five classes in increasing order of TPF. The number transplanted depended on the phenotype frequency. The selection index was the quotient of the number selected and the total number of patients. The selection index was 0.28 to 0.43 to 0.79 to 1.34 to 2.38 from class 1 to 5. To equilibrate the transplantability of rare phenotype recipients on the waiting list, a bonus was introduced for the most disadvantageous frequency class. Adding the bonus modified the selection index as follows: 1.0 to 1.25 to 1.5 to 1.34 to 2.38, which appears more equilibrated except for the class 5. In conclusion, if a bonus is applied for rare phenotypes, the chance to be transplanted becomes similar between patients with other parameters the same. PMID- 15110564 TI - Kidney transplantation from donors aged more than 65 years. AB - BACKGROUND: There are no agreed criteria to predict the outcome of elderly donor kidneys or to decide between single (SKG) or dual (DKG) kidney graft transplantation. METHODS: Between January 1999 and January 2003, 46 SKG and 14 DKG were performed from elderly donors (mean donor age 71.6 years; range: 66 to 87). Kidney biopsies were scored according to Karpinski. A calculated admission creatinine clearance <50 mL/min and/or a biopsy score of 5 or 6 were used to select kidneys for DKG. Grafts with better function or lower biopsy scores were employed for SKG. RESULTS: Mean cold ischemia time (CIT) was 16.8 hours (range 8.1 to 28.6) in SKG, and 16.3 hours (range 4.6 to 24.3) for the first kidney and 17.4 hours (range 5.1 to 25.9) for the second graft in DKG. Delayed graft function (DGF) occurred in 34.1% SKG and in 28.5% DKG. Acute rejection rates were 9.1% for SKG and 0% for DKG. Three-year actuarial patient survival rates were 97.7% for SKG and 92.9% for DKG; for kidneys, 95.4% and 92.9%. One-year mean serum creatinine levels were 1.8 mg/dL (range 1.1 to 4.0) for SKG and 1.2 mg/dL (range 1.0 to 1.8) for DKG (P =.01). CIT longer than 16 hours was related to increased rates of DGF for both SKG (45.4% vs 22.7%) or DKG (42.9% vs 14.3%) and reduced 3-year graft survival rates (SKG: 90.9% vs 100%; DKG: 85.7% vs 100%). CONCLUSIONS: With stringent selection criteria and short CIT (<16 hours), elderly donor kidneys may show good results, thus meaningfully expanding the donor pool. PMID- 15110565 TI - Utilization of suboptimal kidney donors. AB - In recent years over 30% of cadaver donors utilized in the Emilia Romagna (ER) region are over 60 years of age often with some additional risk factors. The organs of these Suboptimal Kidney Donors (SOKD) had been seldomly accepted from 1996 to 1999 (Group 1). From 2000, marking the beginning of use of double transplantation for marginal kidneys, a routine biopsy was performed for SOKD using the Score System proposed by Remuzzi. (Group 2) The 40% of marginal kidneys harvested were utilize in 76% of cases. The few biopsies available for Group 1 patients testify that good kidneys had been discarded, but even in Group 2, when we followed the biopsy Score, strictly several mistakes were suspected: kidneys discarded for an unbelievable difference of 2 points between two kidneys of the same size, cases with an high score based on only a few glomeruli, double kidney transplants that become single and still had fair functions. In the future the utilization of SOKD should probably be improved: (1) with scores only based on a biopsy specimen with more than 25 glomeruli, (2) a score system for incorporating clinical data, and (3) continuous perfusion preservation of kidneys whose characteristics provide some more information that may mitigate some of the damage of marginal donor kidneys. PMID- 15110566 TI - Suboptimal kidney: the experience of a single transplant unit. AB - Not all kidneys are suitable for transplantation. In 2001 in Italy, only 1530 of 1748 organs were transplanted. This difference (-12.5%) not only represents organs harvested from marginal donors considered not suitable, but also kidneys that, although collected from standard donors, had a vascular, parenchymal, or urologic anomaly that made them unsuitable for transplantation. In our center, we established a procedure that defined the characteristics of suboptimal kidneys and allowed us to specifically select the appropriate recipient. We considered as suboptimal all organs with complex arterial anomalies (more than 2 arteries although on a single patch or separated such as to need a double anastomosis or a bench reconstruction); organs with noticeable parenchymal damage (macroscopic sclerosis areas or sutured polar branches accidentally damaged during removal), and organs with complex anomalies of the excretory tract (complete double district). The organs were not considered as suboptimal if they had venous anomalies revised on the bench or if they had a double artery with a single patch <2.5 cm. Such organs were transplanted to recipients who were between 55 and 60 years of age with a body weight of at least 20% less than the donor, and female. PMID- 15110567 TI - Kidney transplants from infected donors: our experience. AB - Organ procurement from infected donors may transmit a disease to the recipient that could cause a graft loss and/or recipient morbidity. Retrospectively, all kidney transplants from infected donors at our center in the last 4 years were reviewed. A donor was considered infected in the presence of at least one positive culture before procurement. From January 1999 to 2003, 23 of 160 donors (14.5%) were infected: in 10 donors a positive blood culture; in 3, a urine culture; and in 13, a bronchial culture. In a further 12 (7%) donors, only the preservation solution was contaminated. Organisms isolated were: Staphylococcus coagulase.neg. (n = 7); Staphylococcus epidermidis (n = 3); Staphylococcus aureus (n = 6); Klebsiella pneumoniae (n = 3); Pseudomonas aeruginosa (n = 4); Acinetobacter (n = 1); Candida albicans (n = 13); Aspergillus (n = 1); and Escherichia coli (n = 1). All except 2 kidneys were transplanted with positivity in all cultures. All recipients received general, nonspecific, antibacterial and antifungal prophylaxis until the antibiotic and antifungal spectrum was ready. Patient and graft survival rates at 6 months were 94% and 93%, respectively. Two deaths occurred due to bacterial arteritis (P aeruginosa), and 2 acute graft losses due to fungal arteritis. Kidneys from infected donors seem suitable for transplants. Only grafts infected by vasculotropic agents (S aureus, P aeruginosa, and C albicans) should be discarded. PMID- 15110568 TI - Medical and surgical complications after kidney transplantation from "suboptimal donors": one centre's experience. AB - To overcome the organ shortage, the pool of donors can be expanded to include aged donors (>55 years old) or patients with diabetes and long-standing hypertension, the so-called "suboptimal donors." Our experience on medical and surgical complications in kidney recipients from such donors and their impact on the graft and patient survival rates is reported. From January 1998 to April 2003, 276 kidney transplantation were performed: 107 from suboptimal donors (group A) and 169 from optimal ones (group B). After a mean follow-up of 26.8 months (range, 1-63 months), the 1-year graft survival rate was 89.3% and 97% for groups A and B, respectively. Medical complications were observed in 18.8% of group A and 6% of group B and surgical complications in 34.5% and 20%, respectively. In conclusion, even if the complication rate is higher among the suboptimal donor group, the patient and graft survival rates appear to be only slightly affected, therefore, validating the use of marginal donors. PMID- 15110569 TI - Sirolimus in kidney transplantation from marginal donors. AB - Nephrotoxicity caused by calcineurin inhibitors can lead to either delayed graft function or long-term decline of renal function after kidney transplantation. Therefore, recipients of renal transplants from marginal donors require non nephrotoxic immunosuppression. Eighteen patients received kidney transplants from marginal donors, with a calcineurin inhibitor-free immunosuppressive regimen, based on basiliximab, mycophenolate mofetil, steroids, and sirolimus. Renal graft biopsy was performed in all cases before surgery. Mean follow-up was 11.8 months. We report immediate renal function in 9 patients, delayed graft function in 5 and acute tubular necrosis in 4 patients. One patient was successfully treated for biopsy-proven acute rejection. Hypercholesterolemia and hypertriglyceridemia were the most common adverse effects (n = 13) associated with arthralgia (n = 2) and thrombocytopenia (n = 2). Five patients underwent a switch to tacrolimus, due to sirolimus-induced side effects. Immunosuppression without the use of calcineurin inhibitors is a safe and effective regimen in kidney transplantation, although sirolimus-related side effects still represent a morbidity factor in these patients. PMID- 15110570 TI - Kidney transplantation from marginal donors. AB - AIM: The increasing demand for transplantation and the shortage of available organs limit the success of organ transplant programs. The use of marginal donors to expand the donor pool is receiving increased attention. We reviewed a 28-month experience of kidney transplants from marginal donors to assess the impact on patient and graft survival. PATIENTS AND METHODS: From January 2001 to May 2003, 78 kidney transplants were performed, including 50 grafts from cadaver donors and 28 from living donors with 3 patients receiving a double kidney transplant. The patients were divided into 4 groups: 31 patients received a kidney from an ideal cadaver donor (group 1a); 19 patients received a graft from a marginal cadaver donor (group 1b); 19 patients received an ideal living related kidney (group 2a); and 9 patients received a marginal living kidney graft (group 2b). RESULTS: Twenty-eight grafts from marginal donors were transplanted with an average follow up of 16 months (range, 1-28 months). The graft survival rates for groups 1a, 1b, 2a, and 2b were 93%, 79%, 100%, and 100% and patient survival rates were 96%, 89%, 100%, and 100%, respectively. CONCLUSION: Despite the observation that use of marginal donors has been associated with a worse outcome compared with ideal donors, we of such grafts resulted in improved quality of life and survival expectancy compared with maintenance dialysis. The marginal kidney donors represent a feasible way to improve the donor pool. PMID- 15110571 TI - Jehovah's witnesses, blood transfusions and transplantations. AB - The firm refusal of blood transfusion treatment by Jehovah's Witnesses has always caused some discomfort to physicians. Two fundamental principles contained in the constitution of our own and other democratic countries specifically dictate that human rights are to be respected. No one may be forced to undergo any particular health treatment, save under the provisions of the law. These principles have led doctors and lawyers to address this difficult patient-doctor issue decisively. On the one hand, a vast jurisprudence has been accumulated, which, although not resolving all the questions regarding the issue, has provided valuable guidelines, elaborated to support physicians in their treatment of patients who refuse homologous blood. On the other hand, wide-ranging research has been undertaken to find therapeutic and surgical methods that make it possible to treat patients without recourse to blood transfusions. Paradoxically, the dilemma presented to doctors by Jehovah's Witnesses has led to new knowledge regarding "the good use of blood," the advantages of which have been experienced also by non-Witnesses. In this article, we briefly consider the roots of Jehovah's Witness beliefs and the principles that lie behind their refusal of blood. Furthermore, we consider which treatments may be accepted, and which are invariably refused. Last, we examine the progress of current scientific research into alternative methodologies and the position of Jehovah's Witnesses when a transfusion is not an option, but a necessity. PMID- 15110572 TI - Erectile dysfunction after kidney transplantation: our 22 years of experience. AB - AIM: To evaluate the results of treatment of erectile dysfunction (ED) in kidney transplant recipients before and after the advent of sildenafil. MATERIALS AND METHODS: From 1981 through 2002, 971 male patients of mean age 53.4 years received a renal graft. Erectile dysfunction (ED) was investigated in all patients at the first urologic visit posttransplantation. Psycho-sexual support was offered to all patients. Before sildenafil use (1998), our diagnostic approach was complex. From 1998 we tested: serum levels of testosterone, prolactin, and glucose with penile duplex ultrasonography and NPT reserved for selected cases. RESULTS: From 1981 through 1998, 365 male kidney transplant recipients (45%) reported ED. Only 169 patients chose to be treated: 27 responded to psycho-sexual therapy; 3 received testosterone with benefit; 133 had a good results from intracavernosal injection of vasoactive drugs; and 6 received a penile prosthesis. Since 1998, 126 patients reported ED (78.3%). Only 78 chose treatment: 24 patients had a satisfactory response to sildenafil (65% with 50 mg and 35% with 100 mg). PGE1 alone or in combination with papaverine and phentolamine produced a good response in 37 patients; 17 patients did not respond to pharmacotherapy; and 5 received a tricomponent penile prosthesis without complications. The side effects of sildenafil and PGE1 therapy were similar to those reported in the literature. CONCLUSIONS: ED is an important problem in male renal transplant recipients. Cultural resistance to treatment is common. However, treatment with sildenafil citrate and intracavernosal self-injection of PGE1 are well accepted, and prosthetic devices may help in resistant cases. PMID- 15110573 TI - Rescue of kidney and pancreas grafts with complex vascular lesions. AB - BACKGROUND: The organ shortage mandates that grafts with complex vascular lesions be considered for graft rescue. METHODS: Surgical graft rescue was attempted in 8 patients bearing 8 kidneys and 2 pancreata that showed complex vascular lesions deemed not suitable for interventional radiology procedures. RESULTS: All procedures but 1 were performed under elective conditions. Seven grafts were repaired in situ, while cooling the organ through retrograde venous perfusion, and 3 kidneys were explanted, repaired extracorporeally, and retransplanted. All vascular reconstructions remain patent after a mean follow-up period of 3.3 years (+/-2.1 years). CONCLUSIONS: Careful patient selection, multidisciplinary evaluation, and personalized surgical technique may allow the rescue of kidney and pancreas grafts with complex vascular lesions that, otherwise, would be lost. PMID- 15110574 TI - Renal vein extension in right kidney transplantation. AB - A short right renal vein may be associated with technical problems in renal transplantation. For this reason, a vena caval extension may be useful to improve exposure of the anastomosis and graft placement. This report evaluates the safety and the effectiveness of renal vein extension, which was routinely performed in right renal transplantation. From April 1986 to December 2002, we performed 371 right kidney transplantations with 252 using the standard technique (group A) and 119 using the renal vein extension (group B). No statistical differences were found between the 2 groups in terms of renal vein thrombosis incidence, delayed graft function, morbidity, and graft loss. Indeed, mean warm ischemia time was reduced in the venoplasty group. In conclusion, renal vein extension is an easy, safe technique that reduces warm ischemia time. We suggest more extensive use of this procedure in right kidney transplantation. PMID- 15110575 TI - Right hemiliver transplants from cadavers or living donors: a comparative analysis. AB - INTRODUCTION: The aim of this article was to compare the results of right hemiliver transplants from living versus cadaver donors in a single institution. METHODS: Between March 1999 and May 2003, we performed 10 right hemiliver transplants from living donors (LD) and 8 right hemiliver transplants from cadavers (CD). The procedure consisted of grafting liver segments 5, 6, 7, and 8. The procedure was performed with a fully perfused liver also in the CD group (in situ split). RESULTS: With follow-up between 7 days and 26 months in the LD group, 2 patients died with functioning grafts: 1 patient died because of massive pulmonary bleeding due to Rendu-Osler Syndrome; the other one died as a consequence of systemic aspergillosis. One patient underwent retransplantation due to arterial thrombosis. In the CD group with a follow-up between 31 days and 48 months, 3 patients died due to sepsis, including 2 who were status 2A. There were 4 early complications among the LD group and 5 in the CD group. The patient and graft survival rates were 80% and 70%, respectively, in the LD group; and both about 62% in the CD group. CONCLUSION: Right hemiliver grafts are at high risk due to technical and septic complications. A higher morbidity is observed in the CD group, where the vascular and biliary tree anatomy cannot be investigated with accuracy. We must avoid transplanting status 2A recipients with this kind of graft. PMID- 15110576 TI - Right hemiliver transplants from living donors: report of 10 cases. AB - INTRODUCTION: A right lobe living related liver transplantation (LRLT) was performed for the first time in Italy on March 16, 2001 at our institution. METHODS: All donors underwent celiac and mesenteric axis angiography. Computed tomography scan to determinate the liver size and anatomical vascular variation, cholangio-magnetic resonance imaging, intraoperative cholangiography, and ultrasonography. All recipients were status 2B on the waiting list for cadaveric liver transplants. The surgical procedures were carried out by grafting segments 5, 6, 7, and 8 of the donor liver. RESULTS: Of the donors, all are alive; 4 had uneventful postoperative courses, 3 had moderate right pleural effusions; 3 had bilious drainage that resolved spontaneously: and 1 had a biliary leak and a pulmonary embolism. Of the recipients, 8 are alive with well-functioning grafts. One recipient has undergone retransplantation due to an arterial thrombosis and another recipient developed a stricture of the biliary anastomosis. Two recipients died: one because of pulmonary hemorrhage in Rendu-Osler syndrome, the other as a consequence of overwhelming systemic aspergillosis. CONCLUSIONS: Our experience suggests that few anatomical vascular and biliary variations are considered contraindications for right lobe LRLT. This challenging surgical procedure seems effective for well-selected recipients of United Network for Organ Sharing II B status. Appropriate recipient selection is crucial as we face a living donor. PMID- 15110577 TI - Split liver transplantation in Italy. AB - The role of split liver transplantation has been well established. The limitation to this technique is the number of potential recipients for a left lateral segment graft. The optimal use of the donor pool is to split the liver to provide 2 grafts suitable for adults obtaining right or left lobe. We explored the potential increase in the number of liver grafts gained from systematically using the technique of splitting on national basis. The crucial factor appeared to be creation of guidelines for the use of optimal livers to optimize organ allocation while minimizing pretransplantation mortality and maximizing post-orthotopic liver transplantation outcome. PMID- 15110578 TI - Donor pool expansion in liver transplantation. AB - INTRODUCTION: The shortage of donors has made it necessary to consider older subjects, those with mild or moderate steatosis, and those who are HBcAb- or hepatitis C virus (HCV)-positive as marginal donors. MATERIALS AND METHODS: From April 1986 to January 2002, 690 orthotopic liver transplantations (OLTs) were performed in 603 patients. In this series we used 68 donors older than 70 years, 51 with steatosis (38 mild, 12 moderate, and 1 severe), 44 were HBcAb-positive and 6 were HCV-positive. RESULTS: Of 68 grafts from donors older than 70 years, 65 were used as a first OLT. These grafts showed 3 PNF, 11 arterial complications, 12 re-OLTs, and 14 deaths with graft survival of 72.3% and 61.34% at 1 and 3 years, respectively. All patients who received the other 3 grafts, which were used for re-OLT, died between postoperative day 21 and 720. Among the 51 grafts with steatosis, we observed 2 PNF of those within the mild steatosis group and graft survival rates of 76.8% and 70.9% at 1 and 3 years, respectively. Forty-four grafts from HBcAb-positive subjects were used in 18 HBsAg-negative and 26 HBsAg-positive recipients. Among the untreated patient group, 1 patient demonstrated hepatitis B virus (HBV) reinfection and 1 patient had de-novo HBV. No reinfection or de novo infections were observed in the 13 patients treated with immunoglobulin or in the 19 patients treated with lamivudine plus immunoglobulin, or in the only patient treated with lamivudine. Graft survival rates were 64.1% and 54.7% at 1 and 3 years, respectively. Among who received 6 patients transplants from HCV-positive donors, we observed 1 recurrence of chronic hepatitis, 1 re-OLT for hepatic vein stenosis, and 1 PNF. CONCLUSION: Old donors, those with moderate steatosis, or those who are HBcAb- and HCV-positive can be safely used in selected recipients to reduce waiting list mortality. PMID- 15110579 TI - Comparison of Celsior and University of Wisconsin solutions in cold preservation of liver from octogenarian donors. AB - BACKGROUND: Celsior (CS) has recently been proposed as a cold storage solution for thoracic and abdominal organs. We compared University of Wisconsin (UW) and CS solutions for the preservation of livers from old donors, with regard to initial function as well as short- and long-term graft and patient survival. METHODS: A multicenter retrospective study from 1998 to 2002 includes 30 livers from octogenarian donors preserved in CS (n = 15) or UW (n = 15) solution prior to transplantation. Donor and recipient clinical and laboratory parameters as well as liver biopsy results were evaluated in all cases. RESULTS: The distribution of the main donor variables as well as recipient characteristics were comparable between groups. Mean cold ischemia time was 421 minutes in the CS group and 474 minutes in the UW group. Mild steatosis was present in 8 cases in the CS group and 7 cases in the UW group. No primary graft dysfunction or arterial or biliary complications were noted. There was 1 acute rejection episode in the CS group and 4 in the UW group. Late postoperative deaths were observed only in the UW group (ie, 7 of 15). Actuarial graft survival was 100% in the CS group vs 86.7% in the UW group (P = NS) at 3 months, and 100% in the CS group vs 52.5% in the UW group (P =.007) at 12 months. Patient survival was 100% in the CS group vs 93.3% in the UW group (P = NS) at 3 months, and 100% in the CS group vs 59.3% in the UW group (P =.01) at 12 months. CONCLUSIONS: Both CS and UW solutions effectively protect livers obtained from donors >80 years of age during the early postoperative course but the CS group had better long-term results. PMID- 15110580 TI - Marginal donors in liver transplantation. AB - Marginal liver donor criteria included the following: obesity (weight >100 Kg or BMI >27), age >50 years; macrovesicular steatosis >50%; intensive care unit stay >4 days; prolonged hypotensive episodes of >1 hour, and <60 mm Hg with high inotropic drug use (dopamine, [DPM] > 14 microg/kg per minute); cold ischemia time >14 hours, peak serum sodium >155 mEq/L; sepsis, viral infections, and alcoholism; high levels of bilirubin, ALT, and AST, or extrahepatic neoplasia. Between August 1992 and May 2003, we performed 251 liver transplants in 241 patients of whom 155 are presently alive. We used 124 (49.4%) standard donors and 127 (50.6%) marginal donors. Among the group that received a standard donor, 81 (65.3%) are still alive. Among recipients of organs from marginal donors. 81 (63.8%) are still alive. We also assessed the quality of donors according to the severity of recipient disease. For standard donors these outcomes were 61.5% for UNOS 1, 37.5% for UNOS 2A, 73.2% for UNOS 2B, and 80% for UNOS 3 for marginal donors they were 46.1% for UNOS 1, 53.6% for UNOS 2A, 70.7% for UNOS 2B, and 63.6% for UNOS 3. Among the patients who received a liver from a donor >60 years old, there were no survivors in UNOS 1 and 2A, but there were good results in groups 2B and 3. These results suggest there is no difference between marginal and standard donors, even in sick patients, with the exception of donor age. PMID- 15110581 TI - Liver transplantation from old donors into HCV and non-HCV recipients. AB - BACKGROUND: Chronic liver failure due to HCV-related cirrhosis is the leading indication for liver transplantation in Western countries. Inferior long-term results have been reported for liver transplantation in HCV patients, especially when marginal donor livers are utilized. The aim of this study was to retrospectively analyze the outcome of liver transplantation from elderly donors in HCV versus non-HCV recipients. METHODS: One hundred seventy-nine patients receiving 204 liver transplantations were divided into four groups according to HCV positivity and donor age (> or <65 years). Long-term survivals were calculated by the Kaplan-Meier method. RESULTS: Grafts from donors of >65 years into HCV-positive patients displayed lower patient and graft survival rates than HCV-negative cases, although macrosteatosis was more frequent (55% vs 9%, P =.02) among organs used for non-HCV cases. Moreover, HCV-positive recipients transplanted with a donor aged >65 years had significantly lower patient and graft survival (40% vs 78% [P =.01] and 40% vs 68% [P =.06], respectively) than patients receiving a liver from a younger donor. CONCLUSIONS: Our retrospective analysis, although hampered by a small number of patients transplanted with an old liver, suggest that the results of liver transplantation with a donor graft >65 years of age into an HCV-positive recipient shows a worse outcome than those from younger donors. Older livers should be reserved for non-HCV cases. PMID- 15110582 TI - Prediction of 6-month survival after liver transplantation using Cox regression. AB - BACKGROUND: The outcome of liver transplantation (LTx) has been correlated with several donor and recipient factors. METHODS: A database of 191 consecutive LTx cases was analyzed using Kaplan-Meier and Cox regression statistics based on 80 variables. To avoid additional effects of late events on patient survival, the chosen endpoint was 6 months. Data were evaluated using SPSS statistical software. RESULTS: Kaplan-Meier analysis revealed a difference in 1- to 6-month graft survival between patients transplanted with organs from donors older versus younger than 60 years (Breslow, P <.01). Differences in 1- to 6-month graft survivals were observed between patients listed as UNOS status 3, 2B, 2A, and 1: the outcomes for UNOS status 2B versus UNOS status 2A and UNOS status 2B versus status 1 were significant (P <.05). Differences in 1- to 6-month graft survival rates were found between patients with versus without sepsis (P <.05), and with versus without rejection episodes (P <.01). Cox regression analysis revealed only three of the variables to be independent prognostic predictors of graft failure: donor age; postoperative septic status; and rejection. The best mathematical multivariate Cox regression model linked donor age + donor Na + rejection + sepsis to 1- to 6-month graft survival (chi-square = 29.06, P <.001). CONCLUSION: Factors predictive of 1- to 6-month graft survival after liver transplantation include donor age; UNOS status; sepsis; and rejection. PMID- 15110583 TI - Potential predictive value of the MELD score for short-term mortality after liver transplantation. AB - In the last years, a model for end-stage liver disease (MELD) was suggested as a disease severity score for patients with end-stage liver disease awaiting liver transplantation. In the early 2002, United Network for Organ Sharing (UNOS) has proposed to replace the current status 2A, 2B, and 3 by a modified version of the original MELD score based upon patient risk for 3-month mortality on the waiting list. In this study UNOS status and MELD score were evaluated retrospectively for postoperative 3-month mortality in patients who underwent liver transplantation from 2000 to 2001. Liver recipients were stratified for UNOS status 2A, 2B, and 3, and the corresponding MELD score was calculated for each patient. A receiver operating characteristic (ROC) analysis was performed for both conventional UNOS status and MELD score by fitting patient deaths within 3 months after liver transplantation. The MELD score revealed a better prediction rate for 3-month mortality after the first LT than conventional UNOS status, although no statistical significance was evident by ROC curve comparison. This preliminary study seems to suggest a potentially better predictive rate for the MELD score than conventional UNOS status concerning short-term mortality after liver transplantation. PMID- 15110584 TI - Low-dose intramuscular hepatitis B immune globulin and lamivudine for long-term prophylaxis of hepatitis B recurrence after liver transplantation. AB - The combination of lamivudine and hepatitis B immunoglobulins (HBIg) to prevent recurrence of HBV hepatitis has significantly improved the survival of patients transplanted for HBV-related end-stage liver disease. Generally, HBIg are administered intravenously. We evaluated the efficacy, tolerability, and cost savings of long-term intramuscular HBIg and lamivudine in 28 patients (23 men and 5 women), who received liver transplants for acute or chronic HBV-related liver disease. Twelve patients started lamivudine before and 16 at the time of liver transplantation. HBIg were administered intravenously during the first week (50 to 70,000 IU) and intramuscularly thereafter (1200 IU every 3 to 6 weeks) to maintain an HbsAb titer >100 IU/L. Mean follow-up was 20 +/- 13 months. Only one patient experienced HBV recurrence (9 months after transplantation). This patient had failed to follow the scheduled prophylaxis. Cumulative survival at 3 years was 83%. Intramuscular HBIg were well tolerated in all cases. Cost analysis comparing intramuscular vs intravenous HBIg administration showed that 39,490 Euros were saved per patient per year. These preliminary results show that low dose intramuscular HBIg and lamivudine are efficacious and cost-effective for long-term prophylaxis of hepatitis B recurrence after liver transplantation. PMID- 15110585 TI - Selection and preparation of candidates for combined liver-kidney transplantation: experience at a single center-two case reports. AB - BACKGROUND: The published experiences of combined liver-kidney transplantation (LKT) are favorable, but there is still no uniformity concerning the impact on hepatorenal syndrome, or in cases of symptomatic hepatorenal polycystic disease. Herein we describe our experience with two LKTs, with particular reference to the selection and preparation of the candidates, and the surgical approach. METHODS: Between 1996 and June 2003, we performed 430 liver transplants in 398 recipients, including two LKTs: one in a patient with hepatorenal polycystic disease (case 1) and the other in a patient with HBV(+) cirrhosis undergoing dialysis after a previous isolated kidney transplant (case 2). RESULTS: In case 1, LKT and right nephrectomy were performed 2 months after a left lumbar nephrectomy. In case 2, LKT was performed 10 months after an isolated kidney transplant, without removing the first graft, which recovered function after 3 months. Both patients are now in good health with functioning grafts. CONCLUSIONS: LKT requires careful selection and preparation of candidates to optimize the probability of success. In well-compensated dialyzed patients with cirrhosis due to viral hepatitis, we believe that a combined approach is indicated after antiviral therapy. In cases of hepatorenal cystic disease, a two-stage surgical approach makes it possible to eliminate the risk of infection and intracyst hemorrhage in nonfunctioning polycystic kidneys. PMID- 15110586 TI - Combined liver-kidney transplantation: experience at Bologna's transplant center. AB - BACKGROUND: The results obtained for combined liver-kidney transplantation (CLKT) are reported. METHODS: From 1997 to 2003 six procedures were performed in patients with polycystic disease (n = 2), chronic glomerulonephritis associated with HCV cirrhosis (n = 2), chronic interstitial nephritis associated with cirrhosis HCV + alcohol (n = 1), and subacute hepatitis caused by drugs (n = 1). The average age of patients was 55 years (5 M and 1 F). The liver transplants were performed using the piggyback technique except in the two polycystic patients, for whom a conventional technique was used. The immunosuppressive therapy was based on cyclosporine in five patients and tacrolimus in one patient. RESULTS: The average blood transfusion was 3670 mL, with larger consumption in the two polycystic cases. Graft function immediately after operation was satisfactory. There were two cases of liver and kidney rejection. In one case, a cardiac arrest occurred during the operation with neurologic consequences. The postoperative complications were cyclosporine neurotoxicity (n = 1), pleuric empyema treated with surgical drainage (n = 1), and CMV infection (n = 1). Four years after CLKT, one patient underwent nephrectomy for a de novo tumor. Of the six patients, four are in good general condition whereas the two polycystic disease patients died within 1 month from transplantation due to ARDS and MOF, respectively. CONCLUSIONS: Based on the worst results of CLKT occurred among patients with polycystic disease, a more rigorous selection is necessary. PMID- 15110587 TI - Sequential liver-kidney transplantation. AB - The indications for sequential liver and kidney transplantation have not been well defined. Two categories of patients may benefit from this procedure: patients with primary renal disease associated with hepatic disorders (glomerulonephritis, tubulointerstitial nephritis, metabolic diseases, and structural diseases) and patients who develop renal failure after liver transplantation. Chronic renal failure is a frequent long-term complication after liver transplantation. End-stage renal disease develops in 2% to 10% of cases by 10 years after transplantation. Kidney transplantation appears to be a better option than dialysis for the treatment of end-stage renal disease after liver transplantation. In contrast, survival rates, after kidney transplantation are significantly lower among liver transplant patients than primary-only kidney transplant recipients. Considering the donor shortage, kidney transplantation should be cautiously considered in liver transplantation patients. New immunosuppressive drugs and protocols are needed to reduce chronic renal failure after liver transplantation. PMID- 15110588 TI - Hepatic resection after liver transplantation as a graft-saving procedure: indication criteria, timing and outcome. AB - BACKGROUND: Hepatic resection is uncommon after liver transplantation (LT), but can be a graft-saving procedure in selected cases. Herein we describe the criteria, outcome, and timing of this procedure in our series. METHODS: Between January 1996 and December 2002, 397 LTs were performed in 367 recipients, of whom 12 patients (3.2%) subsequently underwent liver graft resections because of ischemic-type biliary lesions (ITBLs) (n = 5, 41.6%), segmental hepatic artery thrombosis (S-HAT)(n = 3, 25%), recurrent hepatocellular carcinoma (HCC) (n = 2, 16.6%), liver abscess (n = 1, 8.3%), or liver trauma (n = 1, 8.3%). The patients were divided into group 1 (n = 3 all with S-HAT) who underwent early resections (within 3 months of LT), and group 2 (n = 9) who underwent late resections (after 3 months). The outcomes and postoperative mortality ratio (within 30 days) were compared. RESULTS: The resections consisted of four left lobectomies, three right hepatectomies, two extended right hepatectomies, one segmentectomy, one anterior trisegmentectomy, and one right lateral sectoriectomy. The perioperative mortality rate was 66.6% in group 1 (one case of myocardial infarction and one of sepsis), and 22% in group 2 (one case of sepsis and one of hepatic failure). CONCLUSIONS: Late resections in stable patients with damage confined to the graft yield good prognosis. Even major resections are feasible graft-saving procedures. In contrast, early hepatic resections in S-HAT are associated with a worse outcome. Retransplantation should be considered the first-choice option. Sepsis significantly affects the postsurgical course. PMID- 15110589 TI - Intraabdominal pressure in liver transplant recipients: incidence and clinical significance. AB - BACKGROUND: The incidence and clinical relevance of increased intraabdominal pressure after orthotopic liver transplantation (OLT) has not yet been evaluated despite the finding that occurrence of this condition in postsurgical critically ill patients may impair various organ functions. The aim of this study was to assess whether the occurrence of abdominal hypertension among a population of OLT recipients was an important cofactor producing early postoperative complications. METHOD: This prospective clinical study measured abdominal pressure every 6 hours during the intensive care unit (ICU) stay using the urinary bladder method. A value of >/=25 mm Hg was considered high. Hemodynamic status was simultaneously evaluated and renal function assessed based on the hourly urinary output, and by calculating serum creatinine on postoperative days 2 and 4. Renal failure was defined as a serum creatinine level of >1.5 mg/dL, or an increase in peak of >1 mg/L within 72 hours of surgery. The filtration gradient and patient outcomes were also considered. RESULTS: Intraabdominal hypertension was observed in 32% of cases. The subjects displaying high IAP showed significantly lower artery pressure values (P <.01), but did not differ in terms of central venous pressure or cardiac output. High intraabdominal pressure was more frequently associated with renal failure (P <.01), a lower filtration gradient (P <.001), delayed postsurgical weaning from the ventilation (P <.001), and increased ICU mortality (P <.05). A receiver operator characteristic curve analysis showed that the critical IAP values, namely those with the best sensitivity/specificity, were 23 mm Hg for postoperative ventilatory delayed weaning (P <.05), 24 mm Hg for renal dysfunction (P <.05), and 25 mm Hg for death (P <.01). CONCLUSIONS: Abdominal hypertension occurs frequently after OLT and may be associated with a complicated postoperative course. PMID- 15110590 TI - Interventional ultrasound: experience in 426 orthotopic liver transplantations. AB - BACKGROUND: The purpose of this study was to analyze the role of interventional ultrasound in the treatment of nonvascular complications in liver transplant recipients. METHODS: Between August 1996 and May 2003, we performed 426 OLTs in 394 patients, 287 men (73%) and 107 women (27%), mean age of 50 +/- 9.5 years (range 17 to 68.2). A total of 2556 diagnostic ultrasound examinations were performed, resulting in a mean of 5.9 per patient (range 2 to 21). The interventional maneuvers included: echo-guided biopsies; drainage of abdominal or thoracic effusions; drainage of abdominal, intrahepatic, or splenic collections; positioning of biliary drains; and use of the "rendezvous" technique. RESULTS: Six hundred seventy-seven echo-guided interventional maneuvers were performed in 394 OTL patients, comprising 417 (61.6%) biopsies and 260 (38.4%) therapeutic maneuvers. Eighty-one ascite drains were positioned (31.1%); in 73 cases, pleural effusions were drained (28.1%). Sixty-seven abdominal or intrahepatic collections were drained (25.8%), of which 36 (53.7%) were due to bilomas or biliary peritonitis, 15 (22.4%) hematomas, 4 (5.9%) hepatic abscesses, 11 (16.4%) infected abdominal collections, and 1 (1.5%) splenic abscess. Thirty-nine cases (15%) of biliary drainage were performed. In 33 cases (7.9%), the parenchymal biopsies were not diagnostic because of an inadequate specimen. The treatment success rate was 96.1%. No complications related to the therapeutic maneuvers were recorded, but there were 5 biopsy-related complications (1.2%). CONCLUSIONS: Echo-guided interventional maneuvers are safe, produce a high success rate, and represent an important option in the management of OLT patients. PMID- 15110591 TI - Role of interventional radiology in the management of vascular complications after liver transplantation. AB - The purpose of this study is to review the role of the percutaneous interventional procedures in the treatment of vascular complications after orthotopic liver transplantations (OLT). Vascular complications, such as arterial stenosis and venous thrombosis, which occur in approximately 1% to 10% of liver transplant patients, are associated with a higher risk of graft dysfunction. Percutaneous interventional procedures, including angioplasty, local thrombolysis, and embolization, are useful to manage these complications. A reduced blood loss and a low incidence of procedural complications allow for rapid recovery. Hepatic arterial and portal vein anastomotic stenosis can be treated effectively by means of balloon dilation; stenting has also been proposed, particularly for venous complications. Infusional local thrombolysis may be useful in venous thrombosis. Arteriovenous fistulas, occurring at the level of the anastomosis or after liver biopsy, require intraarterial embolization using microcoils or gelfoam. Timing of the intervention for the treatment of ischemic complications is of outmost importance to guarantee liver functional recovery and avoid irreversible parenchymal injuries. Other interventional procedures may be extremely useful to manage portal hypertension after OLT; for example, by creation of transjugular portosystemic shunts, or, in the case of associated hypersplenism, transarterial embolization of the splenic artery. Finally, in patients with recurrent hepatitis, the transjugular approach has been shown to be safe and effective for liver biopsy, whereas transarterial chemoembolization may be extremely useful to treat recurrent hepatocarcinoma. PMID- 15110592 TI - Role of intensive therapy in liver transplant recipients: experience in blood purification and biosynthetic techniques. AB - The severity of the clinical situation for patients awaiting liver transplant depends on the conditions induced by the liver disease, which trigger a series of extrahepatic system alterations requiring prompt correction to improve the patient's preoperative condition. In the pre-terminal stage of liver cirrhosis, the patient's general condition is partly due to toxemia caused by loss of blood purification functions and also to liver synthetic deficits. In this study we describe our experience with extracorporeal support techniques, both from the blood purification standpoint and from biosynthesis. This experience represents an historic overview of the techniques involved in for construction of an "artificial liver." The aim of this work was to provide a summary of our clinical findings and current methods in extracorporeal liver support with the goal of improving the clinical status potential transplant patients. PMID- 15110593 TI - Successful minimally invasive management of late portal vein thrombosis after splenectomy due to splenic artery steal syndrome following liver transplantation: a case report. AB - Portal vein thrombosis (PVT) after liver transplantation (OLT), which occurs in 1% to 2.7% of cases, can compromise patient and graft survival. Percutaneous transhepatic portal vein angioplasty offers an option to treat PVT, diminishing surgically related morbidity and the need for retransplantation. We describe a case of late PVT after OLT, which was successfully treated by a minimally invasive percutaneous transhepatic approach using both mechanical fragmentation and pharmacologic lysis of the thrombus followed by anticoagulation. The patient has had a good clinical course with normal graft function and patent portal blood flow at 6-month follow-up. This case report confirms the possibility of successful recanalization of the portal vein in a patient with late PVT after liver transplantation. Sustained anticoagulation/antiaggregation therapy for at least 6 months after the procedure is advisable. PMID- 15110595 TI - Pancreas transplantation from marginal donors. AB - BACKGROUND: Marginal donor organs are a supplementary source of grafts that has not been fully exploited for pancreas transplantation (PTx). METHODS: A total of 100 PTx were performed with grafts procured from either 48 nonmarginal donors (NMD) or 52 marginal donors (MD), namely age greater than 45 years and/or severe hemodynamic instability at the time of procurement. PTx outcome was evaluated as the incidence of delayed endocrine pancreas function (DEPF), the complication rate, and the patient and graft survivals. RESULTS: The DEPF rate was 6.2% for NMD as compared to 0 for MD (P >.05). Relaparotomy rate was 12.5% for NMD and 9.6% for MD (P >.05). Actuarial 1-year graft survival was 91.7% and 94.2% for NMD and MD, respectively (P >.05). Equivalent figures for patients were 97.9% and 98.1%, respectively (P >.05). CONCLUSIONS: Pancreas from MD may be safely employed and significantly expand the donor pool for PTx. PMID- 15110594 TI - Pancreas preservation with University of Wisconsin and Celsior solutions. AB - BACKGROUND: Although the use of Celsior has been recently described for heart, lung, liver, and kidney transplantation, no data are available on its use for clinical pancreas preservation. METHODS: We herein describe the results of 112 pancreas transplants preserved with either University of Wisconsin (UW; (n = 56) or Celsior (n = 56) solution at two Italian transplant centers. The groups were comparable with regard to all donor and recipient characteristics. RESULTS: Mean cold and warm ischemia times were 10.1 +/- 2.2 hours and 37.2 +/- 8.2 minutes for UW compared to 10.8 +/- 2.4 hours and 38.3 +/- 6.7 minutes for Celsior (P = NS). Delayed endocrine pancreas function was recorded in two UW-preserved grafts (3.6%). Actuarial 1-year patient survival was 94.6% for UW as compared with 100% for Celsior (P = NS). Equivalent graft survival figures were 91.0% for UW as compared with 96.4% for Celsior (P = NS). CONCLUSIONS: Within the range of cold ischemia times reported in this study, UW and Celsior solutions have similar safety profiles for pancreas transplantation. PMID- 15110596 TI - Pancreas transplant alone. AB - Pancreas transplant alone (PTA) represents a growing proportion of overall pancreas transplantations, with 1-year patient and graft survivals of almost 100% and higher than 80%, respectively. PTA can restore normoglycemia without exogenous insulin administration and eliminate acute diabetic complications. In our series of 28 PTA, performed with portal-enteric drainage, 2-year patient and pancreas survivals were 100% and 87%, respectively. In patients with successful transplantation, rapid normalization of blood glucose level and HbA1c concentration was observed, due to restored endogenous insulin secretion. Several classical cardiovascular risk factors were measured before and after transplant, with significant improvements shortly after transplantation. Diabetic retinopathy improved in 58.8% of examined eyes, stabilized in 35.3%, and worsened in 5.9%. In conclusion, PTA represents a clinically relevant option for patients with type 1 diabetes without advanced renal disease. It restores normoglycemia in the vast majority of patients and seems to have a positive impact on late diabetic complications. PMID- 15110597 TI - Retroperitoneal pancreas transplantation with portal-enteric drainage. AB - BACKGROUND: Portal-enteric drainage (PED) is the latest refinement in the surgical technique for pancreas transplantation (PTx). We herein describe the results of a modified technique for PED that places the pancreas in a totally retroperitoneal position. METHODS: Between April 2001 and June 2003, 79 PTx were performed using a retroperitoneal PED technique. RESULTS: No graft was lost due to surgical complications and the relaparotomy rate was 11.4%. Mean hospital stay averaged 25.9 days (+/-14.4 days) with a 30-day readmission rate of 12.7%. One graft was lost due to delayed (6 months) arterial thrombosis and three to acute rejection. The overall 1-year patient and graft survivals were 98.7% and 93.7%, respectively. CONCLUSIONS: Our data confirm that PED of pancreas grafts is associated with low morbidity and mortality rates. Whether retroperitoneal graft placement has actual advantages over the "classical" intraperitoneal position remains to be ascertained. PMID- 15110598 TI - Small-bowel obstruction due to Bezoar following pancreas transplantation with portal-enteric drainage: a case report. AB - BACKGROUND: Despite recent improvements, surgical complications continue to occur frequently after pancreas transplantation, remaining the leading cause of early graft loss. Small-bowel obstruction, however, is exceedingly rare; it has not been associated with an enhanced risk of graft loss. METHODS: Intestinal obstruction occurred 7 days after pancreas transplantation due to bezoar blockage at the level of the jejunojejunostomy of the Roux-en-Y loop, which had been constructed to drain the exocrine secretions of the pancreas graft. RESULTS: CT scan promptly identified the foreign body and greatly facilitated graft rescue before duodenal rupture or the development of graft pancreatitis. Nineteen months after repeat laparotomy the patient is alive with good pancreatic endocrine function. CONCLUSIONS: In cases of pancreas transplantation with enteric drainage, obstruction of the Roux-en-Y loop may create a totally sealed system that may lead to severe duodenal dilation and eventually to duodenal rupture or graft pancreatitis. PMID- 15110599 TI - Simultaneous cadaver pancreas-living donor kidney transplantation. AB - BACKGROUND: The expansion of the donor pool achieved with living kidney donation (LKD) is particularly beneficial for diabetic patients, who have a worse prognosis during dialysis when compared to other kidney recipients. Simultaneous cadaver pancreas-living kidney transplantation (SPLKTx) merges the advantages of LKD with those of cadaver donation, and may be an attractive alternative to simultaneous pancreas kidney transplantation (SPKTx). METHODS: The outcomes of 18 SPLKTx were compared with those of 33 SPKTx. RESULTS: LKD expanded the donor pool from 33 to 51 (P =.004). Median wait time was shorter for SPLKTx (14 days) than for SPKTx (95 days) (P =.006). The risk for surgical complications was not increased by SPLKTx, as witnessed by relaparotomy rates (SPLKTx: 2/18, 11.1%; SPKTx: 2/33, 6.1%; P >.05). Hospital stay averaged 26.1 +/- 11.2 days for SPLKTx and 27.1 +/- 16.3 for SPKTx (P >.05) with equivalent 30-day readmission rates (SPLKTx: 5.5%; SPKTx: 6.1%); (P >.05). One acute kidney rejection occurred in SPLKTx (5.5%) as compared with four in SPKTx (12.1%); (P >.05). Equivalent rates for the pancreas were 5.5% (1/18) for SPLKTx and 3.0% (1/33) for SPKTx (P >.05). Two-year recipient survival rates were 100% for SPLKTx as compared with 96.9% for SPKTx. Equivalent figures for kidney and pancreas were 80.0% and 84.0% for SPLKTx and 96.9% and 96.9% for SPKTx. CONCLUSIONS: SPLKTx is a valuable alternative to SPKTx. Further development of SPLKTX relies on increased rates of living kidney donation. PMID- 15110600 TI - Preemptive pancreas-kidney transplantation: multidisciplinary follow-up starts too late. AB - BACKGROUND: Preemptive pancreas-kidney transplantation is increasingly considered at early stages of nephropathy in type 1 diabetics. A multidisciplinary approach is required, but referral to the nephrologist is often delayed. OBJECTIVE: To analyze the referral pattern of type 1 diabetics to a dedicated nephrology unit and to test the prevalence of indications for pancreas-kidney transplantation in this population, according to early preemptive criteria (creatinine >/= 2 mg/dL and/or nephrotic syndrome). PATIENTS AND METHODS: The setting of study was the first Italian Nephrology Outpatient Unit dedicated to diabetics during 1991 to 2002. The main biochemical and clinical parameters were analyzed at referral. RESULTS: Ninety type 1 diabetics underwent at least one nephrological visit during the period; 85 had data at referral. The referral pattern was stable: 1991 to June 1996 [22 men, 24 women of median age 36 (18 to 65) years; diabetological follow-up 18.0 (3 to 37) years] and July 1996 to March 2002 [26 men, 18 women median age 40 (18 to 65); diabetological follow-up 21.5 (11 to 36) years]. The main biochemical data at referral were superimposable: serum creatinine: 1.2 (0.6 to 3.2) versus 1.3 (0.6 to 7) mg/dL; proteinuria: 0.9 (0 to 11) versus 1.01 (0.05 to 12.3) g/24 hours. Diabetes follow-up was greater in July 1996 to March 2002 [18 (3 to 37) versus 21.5 (11 to 36) years] suggesting an effect of improvements in diabetic care. At referral 76.6% were macroproteinuric 85.6% had signs of end organ damage other than nephropathy; and 30.6% had indications for pancreas kidney grafting (creatinine >/= 2 mg/dL: n = 6 cases; nephrotic syndrome: n = 10; or both n = 10). CONCLUSIONS: One new frontier of transplantation is the need for early multidisciplinary evaluation of type 1 diabetic patients. PMID- 15110601 TI - Cardiac evaluation for simultaneous pancreas-kidney transplantation and incidence of cardiac perioperative complications: preliminary study. AB - Type I diabetes mellitus (IDDM) is associated with an increased cardiovascular risk, and eligibility protocols for simultaneous pancreas-kidney transplantation (SPKT) are consequently accurate for preoperative cardiovascular assessment. According to our algorithm, coronary angiography in SPKT candidates is indicated for patients not only experiencing previous cardiac events or symptoms, but also those with long-standing diabetes (more than 25 years) and/or age over 45 years. Furthermore, a basal transthoracic echocardiographic exam (TTE) is performed to assess cardiac volumes, left ventricular mass, systolic function, and kinesis. The aims of this study were to evaluate perioperative cardiac morbidity and mortality in 18 SPKT-eligible patients, divided into two groups on the basis of the presence/absence of angiographically evident coronary artery disease (CAD), as well as to assess the impact of left ventricular hypertrophy (LVH) on cardiac complications. Cardiac intraoperative morbidity and mortality and postoperative mortality and major morbidity were absent; minor cardiac morbidity consisted only of silent ischemic ECG alterations, without significant differences between groups, although the incidence seemed to be higher in the CAD-positive population. LVH detected preoperatively by TTE exam also failed to correlate with the incidence of such complications. Selection of SPKT candidates by coronary angiography may have positive effects on perioperative cardiac morbidity and mortality. A larger sample size is needed to give the study statistical power. Medium- and long-term follow-up studies are warranted to evaluate the effects of preoperative selection on survival rates. PMID- 15110602 TI - Simultaneous pancreas-kidney transplantation: short- and long-term results. AB - Simultaneous kidney and pancreas transplantation (SKPT) is the treatment of choice for a majority of type I diabetic patients with end-stage renal disease. With continual refinements in surgical technique and an evolving immunosuppressive arsenal, graft and patient survival have continually improved. The purpose of this study was to evaluate the short- and long-term results of SKPTs performed in 174 recipients from June 1985 to March 2003 including 37 segmental grafts with duct occlusion, 73 whole pancreas transplants with bladder diversion, and 64 whole pancreas grafts with enteric diversion. The series includes 160 cases with systemic drainage and 14 with portal drainage. In the segmental pancreas group, patient survival was 85%, 76%, and 53% with pancreas survival of 67%, 36%, and 15%, and kidney survival of 82%, 63%, and 15%, respectively, at 1, 5, and 10 years. Among the bladder diversion group, patient survival was 94%, 83%, and 73% pancreas survival 72%, 67%, and 65%, and kidney survival 89%, 78%, and 58%, respectively, 1, 5, and 10 years. Among the enter diversion group patient survival was 90% and 90% at 12 and 108 months, pancreas survival 80% and 65%, and kidney survival 85% and 85%, respectively. There were significant differences between curves of survival distribution according to the surgical technique applied for patients (P =.04), pancreas (P =.007), and kidney (P =.005). Based on the results from our study, the short- and long-term prognosis after SKPT is satisfactory, especially compared to the outcomes of long term dialysis among patients with end-stage renal disease caused by type I diabetes. PMID- 15110603 TI - Complete reversal of the nephrotic syndrome after preemptive pancreas-kidney transplantation: a case report. AB - BACKGROUND: Nephrotic syndrome due to diabetic nephropathy is presently considered an indication for pancreas-kidney transplantation even in the absence of severe renal failure. Reversal of the nephrotic syndrome has been reported, but the mechanisms of this effect are unclear. AIM: To describe the renal morphofunctional pattern and the pattern of proteinuria before and after preemptive pancreas-kidney transplantation. METHODS: Methods included quantitative and qualitative assessment of proteinuria as well as renal ultrasound and scintiscan. CASE REPORT: A 42-year-old woman with type 1 diabetes since age 24 had widespread end-organ damage. Renal biopsy (2001) showed a mainly nodular pattern of diabetic nephropathy. Following referral (1999), her serum creatinine ranged from 1.6 to 2.2 mg/dL, with nephrotic range proteinuria (glomerular nonselective, tubular complete). Renal scintiscan revealed bilateral, symmetric, well-perfused kidneys. The functional data before pancreas-kidney graft (February 2003) were: serum creatinine 1.6 mg/dL, creatinine clearance 58 mL/min, serum albumin 2.6 g/dL, proteinuria 9.1 g/d. At hospital discharge (March 2003), the creatinine was 1.2 mg/dL, the creatinine clearance 97 mL/min, the proteinuria 0.676 g/d. Two months later, the creatinine was 1.2 mg/dL and proteinuria 0.421 g/d. A renal scintiscan demonstrated the functional prevalence of the grafted kidney (77% of total function), with vital, almost completely excluded native kidneys (functional contribution, 11.5% each). Proteinuria, ranging from 0.3 to 0.6 g/d, showed a physiological pattern. CONCLUSIONS: Functional exclusion of the native kidneys by renal scintiscan gives morphological support to reversal of the nephrotic syndrome. PMID- 15110604 TI - Solitary pancreas transplantation: preliminary findings about early reduction of proteinuria in incipient or evident diabetic type I nephropathy. AB - OBJECTIVE: Our work was aimed to evaluate the precocious reduction of proteinuria in patients suffering from diabetes mellitus type 1 with incipient and evident nephropathy after isolated pancreas transplantation (PTA). MATERIALS AND METHODS: From December 2000 to March 2003, we followed 24 PTA grafts in 24 patients with diabetes mellitus type 1 (mean age 37.8 years; mean duration of diabetes 24.8 years). The pancreas was transplanted with portal-enteric drainage in 23 patients and systemic-enteric in 1 patient. The immunosuppressive therapy used basilixmab induction and tacrolimus, mycophenolate mophetil (MMF), and low dose steroid maintenance therapy. The renal function, proteinuria, and the glucose metabolic parameters were evaluated before and during the following months after transplant. RESULTS: All patients are alive and twenty-one have a well functioning pancreas with three grafts lost. All patients had persistence of normal renal function. Before transplantation 12 patients displayed proteinuria that was clearly reduced in 11 and gone in three patients, all of whom were insulin-independent. CONCLUSIONS: TPA seems to reduce, and in some cases to regress, the proteinuria associated with early diabetic nephropathy. PMID- 15110605 TI - Multirow CT in the follow-up of pancreas transplantation. AB - PURPOSE: Isolated pancreas or combined kidney-pancreas transplantation represents the only therapeutic weapon for complete resolution of type I diabetes mellitus. The purpose of the present study was to investigate the role of multirow CT in the follow-up of these patients. MATERIALS AND METHODS: Thirty patients who underwent isolated (n = 13) or combined kidney-pancreas (n = 17) transplantation, using systemic-bladder (n = 7) or enteric-portal (n = 23) drainage, were evaluated with multirow CT (Light Speed Plus, GE Medical System). The CT study included unenhanced, arterial, and portal phases of the isolated pancreas, and a urographic phase for combined kidney-pancreas transplants. The acquisition was done with 1.25-mm collimation, 0.6-mm reconstruction interval, and a pitch of 6. A CT scan of the thorax was included if patients were suspected to have pulmonary complications. RESULTS: In all cases it was possible to recognize the surgical technique performed for endocrine and exocrine pancreatic drainage, to evaluate the transplanted pancreas, and to identify possible complications. For example, CT identified thrombosis or stenosis of the arterial graft, partial thrombosis of the venous graft, ectasia of the common iliac artery and arterial graft, dehiscence of the duodenal-bladder anastomosis, infected abdominal collections, thrombosis of the internal iliac vein, and pulmonary infections. CONCLUSIONS: Because multirow CT detects complications it is useful to follow isolated pancreas or combined kidney-pancreas transplants. PMID- 15110606 TI - Kidney and pancreas transplants in Jehovah's witnesses: ethical and practical implications. AB - Jehovah's Witnesses refuse blood transfusions but accept solid organ transplants. Six Jehovah's Witnesses received a kidney and/or a pancreas transplant in our center. After a mean follow-up of 31.4 months (range: 18 to 39) all the recipients are alive and well with functioning grafts. However, 1 month after grafting, one recipient required blood transfusions. Hemorrhage was ruled out and the anemia was attributed to drug-related toxicity. Thus, Jehovah's Witnesses can receive a kidney and/or a pancreas transplant without blood transfusions at the time of surgery. However, lifesaving transfusions may be needed later on, which raises additional and unique medical and ethical issues. PMID- 15110607 TI - Islet transplantation in type 1 diabetic patients. AB - Islet transplantation has been shown to improve overall glucose homeostasis and retard the progression of complications in type I diabetic patients. Also the percentage of recipients achieving complete insulin independence has progressively increased over recent years. An unsolved problem is whether the short-term graft function is secondary to progressive islet exhaustion or to recurrent autoimmunity despite the immunosuppressive therapy. The indications for this procedure remain limited to selected type I diabetic patients. The risks of the immunosuppressive therapy are only proposed to type I diabetic recipients with uncontrolled disease, despite all efforts of the diabetologist and the patient (brittle diabetes), or with a poor quality of life due to unawareness hypoglycemia or severe chronic and progressive complications. PMID- 15110608 TI - An alternative and simple method to consistently prepare viable isolated human islets for clinical transplantation. AB - We describe a method to consistently prepare human islets for transplantation. By combining a simple collagenase digestion method and a density gradient purification system, we were able to obtain successful isolations (>/=200,000 islet equivalents, >/=50% purity) in 69% of processed glands. No reagent of animal source was used. Isolated islets were morphologically well maintained and functionally competent, with sterility confirmed in 97% of cases. Two patients were transplanted with islets prepared by this method; graft function was demonstrated for a few months. Improved simplicity and consistency, together with adequate quality of the preparations, are the main features of this isolation method. PMID- 15110609 TI - In vitro modulation of monocyte chemoattractant protein-1 release in human pancreatic islets. AB - Islet transplantation is a new approach to treat type 1 diabetic patients. Despite its great potential and progressively increasing success rate, islet engraftment still represents an unsolved problem. Only part of the transplanted beta-cell mass survives after infusion due to hypoxia and inflammatory reactions, principally mediated by macrophages. We have demonstrated that human islets release monocyte chemoattractant protein-1 (MCP-1), one of the most powerful macrophage chemokines, which may impair the fate of a transplant. In this study we have attempted to modulate in vitro MCP-1 release by human islets. Human islets isolated using the automated method were cultured in CMRL or M199 standard culture media alone or supplemented with (1) two intracellular kinase inhibitors (10 micromol/L RO8220, a protein kinase C inhibitor, and rcAMP 20 micromol/L, a protein kinase A inhibitor) or (2) two antioxidant and cell-protective agents (vitamin E, vitamin B); or (3) immunosuppressive drugs (0.001 to 10 ng/mL cyclosporine, 0.1 to 100 ng/mL rapamycin, 0.1 to 10 ng/mL tacrolimus, 0.001 to 10 ng/mL mycophenolate acid). We observed that the only culture condition that significantly decreased MCP-1 in human islets were CMRL (31 +/- 12 in CMRL vs 539 +/- 184 pg/mL, in M199, P <.05) or cyclosporine (514 +/- 83 pg/mL in control islet vs 307 +/- 13, 231 +/- 44, 192 +/- 4, 242 +/- 113, 169 +/- 15 pg/mL in islet plus cyclosporine ranging from 0.001 to 10 ng/mL, respectively, P >.05). The capacity of in vitro factors to decrease human islet MCP-1 release suggests strategies to increase the success of islet transplantation. PMID- 15110610 TI - Neonatal pig pancreatic duct-derived insulin-producing cells: preliminary in vitro studies. AB - Neonatal pig pancreata could represent an ideal tissue resource for donor islets for transplantation trials. Because functional islet beta-cells could derive from precursors situated in the ductal system, and neonatal animals are better suitable than adults for recovering such elements, we have examined whether isolated neonatal pancreatic ducts (NPD) could form insulin-producing cells. NPD, retrieved from the pancreas by collagenase digestion, were cultured for 2 weeks. A compact tissue monolayer detached by trypsin was re-incubated to form upon culture. The primary tissue monolayer was plated, yielding secondary monolayers that were supplemented in culture with the following factors: insulin transferrin selenium, niacinamide, keratinocyte growth factor, and high glucose, which promoted formation of islet cell-like clusters during 30 days of culture. Upon reaching 50 to 100 microm in diameter, the cell clusters were subjected to morphologic examination (assessment of viability by staining with ethidium bromide+fluorescein diacetate [EB+FD]; staining for insulin with diphenylthiocarbazone [DTZ]); DNA assay; insulin radioimmunoassay both in the basal state and after in vitro static incubation with high glucose; immunolabeling with anti-insulin fluorescent antibodies. Of the cell clusters, 80% were composed of viable cells that faintly showed DTZ staining. Basal insulin was 16.7 microU/mL, but no insulin response was elicited by stimulation with high glucose. Acid-ethanol extraction showed high insulin levels in the clusters. Finally, immunofluorescence for insulin was positive, indicating the presence of beta-cell-like committed elements. In conclusion, NPD may differentiate into insulin-producing cells, which are at a very early stage when the glucose-sensing apparatus is still immature. PMID- 15110611 TI - Identification of in vitro parameters predictive of graft function: a study in an animal model of islet transplantation. AB - The quality of human islets is one of the factors decisive for the success of human islet transplantation. Several parameters have been proposed to characterize islet quality, but none of them has been able to predict the fate of a transplant. The aim of our study was to correlate a panel of in vitro parameters for islet viability with their in vivo function after transplantation in nude mice. Islets were obtained after enzymatic digestion of a human pancreas; they were purified from exocrine tissue using a continuous-density gradient. Two aliquots of islets (1000 and 2000 islets) were transplanted under the kidney capsule of diabetic nude mice. The animals were followed for 1 month with repeated measurements of blood glucose and body weight. One month after transplantation, mice were killed and their graft harvested for histologic analysis. In parallel we studied in vitro islet viability with propidium iodide and fura-2, their insulin content, their purity, and their insulin response to glucose upon static incubation. Ten islet preparations were transplanted: 3 out of 10 preparations did not restore normoglycemia; 4 out of 10 normalized glycemia only in mice receiving 2000 islets, and 3 out of 10 fully restore normoglycemia in all mice. The purity of preparations (R(2) = 0.63 and 0.85, respectively, with 1000 and 2000 islets) and the insulin content (R(2) = 0.75 with 2000 IE) correlated with transplant success. These data show that purity of islet preparations and their insulin content should be useful parameters for the selection of islet preparations for transplant purposes. PMID- 15110612 TI - Donor organ preservation in high-risk cardiac transplantation. AB - AIM: To evaluate safety and efficacy of blood cardioplegia in a retrospective selected (but not randomized) donor/recipient population as standard organ preservation technique in high-risk heart transplants (HTX). MATERIALS AND METHODS: The rationale of different strategies was based on both donor and recipient evaluations. Unstable donors with a long history of well-known risk factors and/or long-distance retrieval were given blood cardioplegia, particularly for HTX candidates in poor preoperative clinical condition. Organ protection was performed by administration of St Thomas II crystalloid cardioplegia in 74 patients (group 1) while 58 others (group 2) received blood cardioplegia. RESULTS: Groups I versus II shows comparable results for immediate postoperative mortality rates (4% vs 7%, P =.4), high doses of inotropic drug support (48% vs 20%, P =.08), and the need for postoperative mechanical assistance devices (9% vs 4.5%, P =.4). In contrast statistically significant differences were observed for occurrence of acute right ventricular failure (50% vs 5%; P =.004), atrioventricular conduction disturbances (63% vs 10%, P =.003), spontaneous sinus rhythm recovery (18% vs 64% P =.0038) and reperfusion interval (RI) (time between removal of aortic cross-clamp and discontinuation of extracorporeal circulation (ECC)) exceeding 30 minutes (70% vs 21%, P =.0004). Higher peak creatine kinase MB mean value (176 +/- 23 vs 90 +/- 19, P =.06) indicated more severe ischemic damage among G1 patients. CONCLUSION: This study suggests that high-risk heart transplant candidates benefit from blood cardioplegia, due to the reduced incidence of both right ventricular failure and severe cardiac arrhythmia. Potential limitations to this novel technique may be linked to the higher expenses due to the need for a perfusion technician. Improved myocardial protection can be seen even in a longitudinal study on chronic rejection: this form of allograft protection may preserve the matrix and the endothelium. PMID- 15110613 TI - Mechanical circulatory support in severe heart failure: single-center experience. AB - Ventricular assist devices (VADs) have become important therapeutic tools to treat patients with end-stage cardiac failure. VADs are an essential component of transplantation programs as they successfully bridge individuals who would otherwise die. Recently left ventricular VAD (LVAD) therapy has been proposed as alternative to heart transplantation (HTx) for patients who are not transplant candidates. Other indications have now expanded into areas such as postcardiotomy failure, acute myocarditis, and acute massive myocardial infarction. From 1988 to May 2003, 80 patients received left or biventricular mechanical circulatory support including 78 as a bridge to and two as an alternative to HT. All patients survived the operation. Mean duration of VAD support was 77 +/- 150 days. Fifty one points (63.8%) underwent heart transplantation; 3 (3.8%) recovered and were weaned from VADs. Major bleeding episodes occurred in 11 patients (13.8%) and major neurologic events occurred in 8 (10%). Sixteen patients (20%) were discharged home while waiting for HTx. Twenty-two patients (27.5%) died on VAD. In conclusion, VAD therapy proved effective in bridging patients with end-stage heart failure to HTx. While on LVAD support patients who were assisted with implantable wearable devices could be discharged at home, improving their quality of life. PMID- 15110614 TI - Impella recover 100 microaxial left ventricular assist device: the Niguarda experience. AB - BACKGROUND: The Impella Recover 100 (IR100) is an intravascular microaxial blood pump used to support blood circulation for a maximum of 7 days in cases of reduced left ventricular function, for example in postcardiotomy low output syndrome or in cardiogenic shock after acute myocardial infarction. MATERIALS AND METHODS: We supported five patients with the IR100. The mean age, cardiac index (CI), and ejection fraction (EF) of our population were 42 years, 1.83 L/min/m(2), and 20%, respectively. Two patients (group A) with ischemic dilated cardiomyopathy were bridged to heart transplant. Two patients (group B) with fulminan myocarditis and septic shock were bridged to recovery. One patient, with severe valvular cardiomyopathy who underwent aortic valve replacement and mitral valve annuloplasty, was supported to weaning from ECC. RESULTS: Mean support time was 9.8 +/- 2.3 days. Only one acute myocarditis patient died from a severe vasoplegic syndrome despite maximal inotropic and vasoactive support. Both group A patients were successfully transplanted. Among group B, the second patient resolved the septic status and was slowly weaned from the device and discharged home with moderate improvement of LV function (EF = 40%). Patient C was weaned from the IR100 and electively placed on the heart transplant recipient list. CONCLUSIONS: IR100 is a device that in our experience can be utilized for various indications for short-term support. In compromised patients where a traditional LVAD is contraindicated, the IR100 showed good results, for it is minimally invasive and does not need ECC or systemic anticoagulation. PMID- 15110615 TI - Evolving practice patterns in heart transplantation: a single-center experience over 15 years. AB - This analysis is a retrospective characterization of evolving patterns in donor and recipient risk factors for early and late outcomes (survival and freedom from rejection) along with determinants of hospital and 1-year mortality after heart transplantation over a 15-year experience in a single center. Profiles and outcomes were evaluated for procedures performed between 1988 and 1995 (group A, n = 105) versus 1996 and 2003 (group B, n = 218). The following parameters were considered: pretransplant diagnosis, recipient age UNOS status, donor age, total postretrieval ischemic time, donor/recipient size match, and degree of myocardial necrosis at biopsy. Recipients in group B were significantly more compromised as demonstrated by UNOS status (11.4% vs 19.3%; P =.05) and pretransplant pulmonary vascular resistance (2.3 +/- 1.5 vs 3.1 +/- 1.5; P =.04). Marginal donors were more frequently used for group B procedures (21.9% vs 47.7%; P <.0001). Outcomes were significantly more favorable among group B patients in terms of hospital mortality (18.1% vs 10.6%; P =.046), and 1- and 5-year actuarial survival (72.4% vs 83.4%, 60% vs 73.3%, respectively; P =.006). Analysis of the causes of death disclosed a significant reduction in fatal events due to graft failure and acute rejection in group B. No difference emerged with regard to actual freedom from acute rejection. Determinants of hospital mortality were pretransplant diagnosis, UNOS status, donor age, and cardioplegic solution. Transplant era, recipient age, infectious episodes, and ischemic necrosis at biopsy were risk factors for 1-year mortality. We conclude that despite extensive usage of marginal donors and selection of worse candidates, significantly better outcomes were achieved due to improvements in global management strategies. PMID- 15110616 TI - Midterm results of a prospective randomized comparison of two different rabbit antithymocyte globulin induction therapies after heart transplantation. AB - This prospective randomized study compared the effects in heart transplant recipients of thymoglobulin and ATG, two rabbit polyclonal antithymocyte antibodies available for induction therapy. Among 40 patients (29 men and 11 women, mean age: 40.7 +/- 14 years) undergoing orthotopic heart transplantation, 20 were randomly allocated to receive induction with thymoglobulin (group A) and 20 to ATG-fresenius (group B). Comparisons between the two groups included early posttransplant (6 months) incidence of acute rejection episodes (grade >/= 1B), bouts of steroid-resistant rejection, time to first rejection, survival, graft atherosclerosis, infections, and malignancies. The study groups displayed similar preoperative and demographic variables. No significant difference was found with regard to actuarial survival (P =.98), freedom from rejection (P =.68), number of early rejections > 1B (P =.67), mean time to first early cardiac rejection (P =.13), number of steroid-resistant rejections (P =.69). Cytomegalovirus reactivations were more frequent among group A (65%) than group B (30%; P =.028). New infections due to cytomegalovirus occurred only in group A (four patients; 20%; P =.05). No cases of malignancies were observed at a mean follow-up of 32.8 +/- 8.9 months. Although thymoglobulin and ATG showed equivalent efficacy for rejection prevention, they have different immunological properties. In particular, thymoglobulin seems to be associated with a significantly higher incidence of cytomegalovirus disease/reactivation. PMID- 15110617 TI - Heterotopic heart transplantation: a single-centre experience. AB - INTRODUCTION: Orthotopic heart transplantation (OHTx) represents the therapy of choice for end-stage heart disease not treatable with medical or conservative surgical approach. Heterotopic heart transplantation (HHTx) is a surgical procedure in which the graft is connected to the native heart in a parallel fashion and it was especially employed in precyclosporine era. The aim of this paper is to present our experience with HHTx. METHODS: From November 1985 till May 2003, 713 heart transplanted patients included 12 (1.7%) received HHTx. Eleven were male, mean age was 50.7 +/- 5.8 years. Five patients suffered from dilated cardiomyopathy and seven from ischemic cardiomyopathy. Indication for HHTx was: a body size mismatch in 11 cases and availability of a marginal organ in one case. RESULTS: Mean ischemic time was 149 +/- 48 minutes and mean cross clamp time was 82.3 +/- 19.1 minutes. In four cases left ventricle aneurysm resection was associated with HHTx. Hospital mortality was 8.3% (one patient due to multiorgan failure). The actuarial survival rates were 92% and 64% at 1 and 5 years, respectively. The causes of death were: liver cancer, liver cirrosis, aortic dissection, cerebrovascular accident, and chronic rejection. CONCLUSIONS: In our experience, HHTx survival rate is comparable to OHTx. Because of the scarcity of donors, use of an undersized or marginal graft is a valid option to increase the number of transplanted patients. The major disadvantages of HHTx are the need for anticoagulant therapy, the more difficult hemodynamic and immunologic follow-up, and the presence of the diseased native heart. PMID- 15110618 TI - Risk factors for smoking abuse after heart transplantation. AB - Patients (n = 103) were studied before heart transplantation with regard to smoking habits by means of a clinical interview, and 81 were submitted to Minnesota Multiphasic Personality Inventory (MMPI). After a mean time of 50.8 +/- 24.2 months from transplant, they were once again interviewed to ascertain their smoking habits after intervention. Nonsmokers (35 of 103) were still nonabusers. Of the remaining 68 patients who ceased smoking before heart transplant, 12 (17.6%) had returned to tobacco abuse. Dividing these 68 patients into two groups based upon the length of smoking cessation before heart transplant (less than 1 year: short term [ST] more than 1 year: long term [LT]), we noticed that the ST group showed a much greater rate of reabuse (8 of 20, 40%) than the LT group (4 of 48, 8.3%, P =.006). Analyzing six scales of MMPI, we found a statistically different score for self-control ability (scale K) in ST and LT smokers compared to nonsmokers (45.5 and 45.5 vs 51.2, P =.026), and for difficult adaptation (scale Ma) in ST compared both to LT smokers and nonsmokers (ST 57, LT 50.5, NS 47.6; P =.042 LT vs ST, P =.0005 ST vs NS). We concluded that patients who have recently decided to stop smoking and show after MMPI compilation a score of >50 for K and <50 for Ma scale have a higher risk of reabuse and need a greater effort by the transplant team to reinforce their will to stop smoking. PMID- 15110619 TI - Heart transplantation in patients with amyloidosis: single-center experience. AB - INTRODUCTION: Amyloidosis is a systemic disease. Heart transplantation in this subset of patients is contraindicated by the majority of authors. In our center, patients with heart failure due to amyloidosis have been evaluated for cardiac transplantation since 1991. The aim of this study was to analyze the outcome of these patients waiting for transplant and the effectiveness of this therapy. MATERIALS AND METHODS: Since 1991, eight patients affected by amyloidosis have been evaluated and enrolled on the waiting list for transplant: five affected by AL lambda type; two by APO A1; and one by TTR. Four were transplanted, three died waiting for a donor (two from cardiac failure, one from sudden death), and one has been recently transplanted after 17 months on waiting list. RESULTS: Since 1985, 713 patients underwent heart transplantation in our center, five of whom were affected by amyloidosis (0.7%). Two are still alive (60 and 41 months) without evidence of cardiac amyloidotic infiltration. One patient recently underwent a combined heart-liver transplantation. Two patients died after the intervention: one sudden death after 23 months with amyloidotic infiltration of transplanted heart, and one multiple organ failure (MOF) due to progression of the systemic disease. CONCLUSIONS: Despite the small size of the group preventing us from drawing definitive conclusion, heart transplantation may prevent therapy to arrest organ damage in patients with isolated cardiac involvement. Cardiac events are the main cause of death. Patients must be followed-up for evolution of systemic disease. The midterm survival is encouraging. PMID- 15110620 TI - Combined heart and liver transplantation in four adults with familial amyloidosis: experience of a single center. AB - There are few reports of combined heart and liver transplantation (CHLT) for familial amyloidotic polyneuropathy (FAP). The technique for the operation remains to be defined. Four CHLTs were performed for amyloidogenic transthyretin related (variant Glu89Gln-ATTR Glu89Gln) cardiomyopathy in our center. Patients 1 and 4 had no serious involvement of other organs, whereas patients 2 and 3 had evident peripheral neuropathy and gastrointestinal motility alterations. Patient 3 also had high-grade orthostatic hypotension. All four patients underwent cardiac and sequential hepatic transplantation with organs procured from the same donor. Venovenous bypass was used in patients 1 and 4 who experienced uncomplicated procedures. The amyloidotic liver of patient 4 was successfully utilized for a domino procedure to treat a patient with hepatocellular carcinoma on cirrhosis. The cardiac performance of patients 1 and 4 remains normal; there has been no progression of amyloidosis at 42 and 1 months after transplantation. Patient 2 had no intraoperative complications but experienced postoperative bleeding, renal failure, sepsis, and heart failure, and finally died of multiorgan failure 2 months after transplant. In patient 3, right hemicolectomy was required intraoperatively due to intestinal ischemia, without significant hemodynamic instability, while extracardiac symptoms of amyloidosis gradually worsened postoperatively. In conclusion, CHLT for ATTR Glu89Gln may be performed even in patients with advanced disease. However, the most compromised patients are more likely to display intraoperative risks, postoperative complications, and worsening of extracardiac, extrahepatic symptoms. PMID- 15110621 TI - Lung transplantation for cystic fibrosis. AB - BACKGROUND: Lung transplantation is a robust therapeutic option to treat patients with cystic fibrosis. PATIENTS AND METHODS: Since 1996, 109 patients with cystic fibrosis were accepted onto our waiting list with 58 bilateral sequential lung transplants performed in 56 patients and two patients retransplanted for obliterative bronchiolitis syndrome. RESULTS: Preoperative mean FEV(1) was 0.64 L/s, mean PaO(2) with supplemental oxygen was 56 mm Hg, and the mean 6-minute walking test was 320 m. Transplantation was performed through a "clam shell incision" in the first 29 patients and via bilateral anterolateral thoracotomies without sternal division in the remaining patients. Cardiopulmonary bypass was required in 14 patients. In 21 patients the donor lungs had to be trimmed by wedge resections with mechanical staplers and bovine pericardium buttressing to fit the recipient chest size. Eleven patients were extubated in the operating room immediately after the procedure. Hospital mortality of 13.8% was related to infection (n = 5), primary graft failure (n = 2), and myocardial infarction (n = 1). Acute rejection episodes occurred 1.6 times per patient/year; lower respiratory tract infections occurred 1.4 times per patient in the first year after transplantation. The mean FEV(1) increased to 82% at 1 year after operation. The 5-year survival rate was 61%. A cyclosporine-based immunosuppressive regimen was initially employed in all patients; 24 were subsequently switched to tacrolimus because of central nervous system toxicity, cyclosporine-related myopathy, or renal failure, obliterative bronchiolitis syndrome, gingival hyperplasia, or hypertrichosis. Ten patients were subsequently switched to sirolimus. Freedom from bronchiolitis obliterans at 5 years was 60%. CONCLUSIONS: Our results confirm that bilateral sequential lung transplantation is a robust therapeutic option for patients with cystic fibrosis. PMID- 15110622 TI - Efficacy and limitations of preemptive therapy against cytomegalovirus infections in heart transplant patients. AB - OBJECTIVES: Cytomegalovirus (CMV) disease often represents a serious complication that promotes opportunistic infections in heart transplant recipients. In this study we evaluated the impact of preemptive gancylovir therapy, guided by pp65 antigenemia on the morbidity associated with viral reactivation. PATIENTS AND METHODS: We have performed a CMV infection surveillance program since March 1999, with antigenemia pp65 determinations weekly for the first 2 months biweekly in the third months, and monthly to the sixth month. Patients with pp65 antigenemia value >/= 10 positive cells per 2 x 10(5) polymorphonuclear cells (PMN) were treated with intravenous gancyclovir followed by 1 month of oral gancyclovir. RESULTS: Among the 107 patients who underwent the virological monitoring, 80 were pp65 antigenemia-positive with preemptive therapy administered in 48 cases. Five patients displayed symptomatic CMV disease (4.7% vs 18% rate in the period of 1988 to 1998 before the introduction of virologic monitoring; P <.01). We observed only one case of gancyclovir-resistant pneumonia which was successfully treated with foscarnet. CMV recurrence in 10 patients required a second cycle of gancyclovir treatment. Our experience included 13 opportunistic infections (12.7%) with 11 antigenemia-positive. CONCLUSIONS: Preemptive therapy drastically reduces the incidence of CMV disease and the associated morbidity. Compared to universal prophylaxis, this approach may avoid unnecessary pharmacologic treatment in more than 50% of transplant recipients. Indeed, preemptive therapy does not fully prevent CMV disease, because it may manifest at the first antigenemia determination, and furthermore may select gancyclovir-resistant strains. PMID- 15110623 TI - "Twinning procedure" in lung transplantation: influence of graft ischemia on survival and incidence of complications. AB - The limited number of suitable lung donors is the major obstacle to clinical application of lung transplantation. The "twinning procedure" may represent one strategy to optimize the use of the small pool of available grafts. From November 1991 to May 2003, 99 single lung transplants (SLTx) were performed including 46 (46%) cases of the "twinning procedure." We divided the study population into two groups: group A (recipients of the "first" lung) and group B (recipients of the "second" lung). The ischemia time was significantly different (A: 216 +/- 48 minutes, B: 310 +/- 89 minutes, P <.001). Differences were not observed in the incidence of graft failure (A: 2, B: 0, P = NS), in the length of mechanical ventilation (A: 12.8 +/- 29.4 days, B: 7.8 +/- 15.2 days, P = NS), or ICU stay (A: 18.8 +/- 50.6 days, B: 15.2 +/- 17.1 days, P = NS), or of hospitalization (A: 37.8 +/- 56.8 days, B: 31.4 +/- 31.7 days, P = NS). Three bronchial anastomotic complications occurred in each group. The incidence of infections (A: 0.015 events/patient/month, B: 0.011 events/patient/month, P = NS) and of treated acute rejections (A: 0.011 events/patient/month, B: 0.011 events/patient/month, P = NS) was similar in the two groups. One-year survival rates were 86% +/- 7% and 72% +/ 10% in group A and B patients, respectively (P = NS). In our experience the different ischemia times related to the twinning procedure did not increase the mortality or morbidity in the early and midterm period. PMID- 15110624 TI - Italian guidelines for intestinal transplantation: potential candidates among the adult patients managed by a medical referral center for chronic intestinal failure. AB - In 2002, the Italian guidelines for eligibility of patients for intestinal transplantation (ITx) were defined as: life-threatening complications of home parenteral nutrition (HPN), lack of venous access for HPN, locally invasive tumors of the abdomen, Chronic intestinal failure (CIF) with a high risk of mortality, primary disease-related poor quality of life (QoL) despite optimal HPN. Our aim was to identify potential candidates for ITx according to these national guidelines among patients managed by a medical referral center for CIF. Records of patients who received HPN were reviewed. CIF was considered reversible or irreversible (energy by HPN <50% or >50% basal energy expenditure). Patients with irreversible CIF were considered eligible for ITx in the absence of a contraindication, as are used for solid organs Tx. From 1986 to 2003 among 64 patients who met the entry criteria 23 showed reversible and 41 irreversible, CIF. Twenty-one patients with irreversible CIF had an indication for ITx, but eight had also contraindications; thus 13 were eligible, including intestinal pseudo-obstruction (n = 6), mesenteric ischemia (n = 3), Crohn's (n = 2), radiation enteritis (n = 1), and desmoid (n = 1). Indications for ITx included HPN liver failure (n = 2), lack of venous access (n = 2), CIF with high risk of mortality (n = 3), very poor QoL (n = 6 including 5 with pseudo-obstruction). According to the Italian guidelines for ITx, 31% of patients with irreversible CIF managed by a medical referral center were eligible for ITx. Primary disease related poor QoL was the indication in half of them. Studies on the QoL after ITx are required to allow patients to make an educated decision. PMID- 15110625 TI - Corneal explantation activity in 2002: an analysis of the modified operational schedule. AB - After data have been gathered about corneal explants performed within the AOP health-care web throughout 2001, the resulting findings were used to update the selection system for donation fitness and operational procedures. The rejection of anti-HBc-positive grafts and tissues coming from subjects more than 79 years old resulted in decreased donations (256 donations, that is 492 corneal explants in 2001 vs 140, that is 273 in 2002), although the number of deaths was unchanged (1298 in 2001 vs 1294 in 2002). Corneas fit for transplantation did not change in number-126 (25.6% of the total available) in 2001 and 113 (41.4%) in 2002-while the instances of rejected corneas occurred 56.3% less frequently, allowing a savings of great deal of human and money resources. After activity schedules were modified, the results analysis confirmed the expected improvement in 2001. PMID- 15110626 TI - Human hand transplantation: what have we learned? AB - Hand transplantation may become an important procedure for upper limb functional restoration. To date, 18 patients have been undergone 24 hand operations in the world. Initial results are extremely promising; the functional results are apparently superior to those obtained with prostheses. We report on the combined French and Italian experience of six patients (eight hands), which is based on a jointly devised protocol and represents the largest available clinical series. Six male patients aged 43, 33, 35, 32, 33, and 22 years received either a single right hand-dominant transplantation (four cases) or a simultaneous double hand transplantation (two cases). The time since the amputation ranged from 3 to 22 years. The level of transplantation was at the wrist in five cases (six hands) and at the distal forearm in two cases (two hands). Cold ischemia averaged 11.5 hours. Three patients simultaneously received additional full-thickness skin taken from the donor and transplanted onto their left hip area. This skin served as a source for biopsies and as an additional area to monitor rejection (distant sentinel skin graft). The immunosuppressive protocol included polyclonal antibodies (three patients) or monoclonal anti-CD 25 antibody (three patients), tacrolimus, mycophenolate mofetil, and prednisolone. No surgical complications occurred. Skin rejection occurred at least once in all patients at a mean of 40 days postoperatively. Three patients recovered protective and some discriminative sensation in their palm and fingers. Two patients are recovering sensation, but are still in the early phases of the regenerative process, due to the short time since the transplantation. One patient was not compliant with the immunosuppressive therapy, and underwent uncontrolled rejection and reamputation. PMID- 15110627 TI - Experimental limb transplantation, part I: identification of an effective tapered triple combination immunosuppressive regime. AB - Limb transplantation was performed across the Brown Norway to Fischer 344 histocompatibility barrier in rats to evaluate the effects of triple combination immunosuppressive therapeutic regimens. Sixty rats were divided into five groups: group I (F344 to F344) isograft controls group II (BN to F344) allograft controls received no immunosuppressive treatment. Groups III and V (BN to F344) received various exposures to tacrolimus (TRL), mycophenolate mofetil (MMF) and prednisolone (Pred) for two weeks: namely, group III: TRL 0.5 mg/kg/d; MMF 10 mg/kg/d; Pred 0.5 mg/kg/d; group IV: TRL 2 mg/kg/d, MMF 15 mg/kg/d, Pred 0.5 mg/kg/d; and group V: TRL 3 mg/kg/d; MMF 20 mg/kg/d; Pred 0.5 mg/kg/d. After 2 weeks, group III and V animals underwent a simultaneous 20% taper of Pred and MMF each further week such that by week 7 the animals were only on TRL. At this time TRL was tapered at the same rate (20% every week) to a maintenance dose of 0.6 mg/kg/d. Evidence of rejection was sought by daily visual observation for swelling, redness, erythema, edema, or skin necrosis. Salvage treatment was used only if rejection occurred after the first 7 weeks, namely, reversing to 100% of the initial TRL dose in that group for 2 weeks with a subsequent taper. Skin and muscle biopsies were obtained from grafted limbs on day 3, 13, 24, 35, and at the endpoint (9 months or uncontrollable rejection). There was no rejection in group I, while all animals showed acute rejection as expected in group II. All group III rats displayed a similar though delayed acute rejection, showing that the regimen was not therapeutic. Rats in group IV displayed the best results, namely, 10 of 12 (83%) with no rejection or side effects at 9 months. Rats in group V displayed numerous, unacceptable side effects due to overtreatment with a 1-month mortality rate of 50%. This study shows that low-dose TRL in combination with MMF and Pred may achieve excellent long-term results of composite tissue transplants. TRL can be used alone as maintenance therapy following an initial loading dose and a tapering period. Rejection is easily reversed by only temporarily increasing the TRL dose. PMID- 15110628 TI - Experimental limb transplantation, part II: excellent return of function and indefinite survival after withdrawal of immunosuppression. AB - In this study the three components of an immunosuppressive combination therapy were gradually withdrawn in a rat limb transplantation model to evaluate the effects on long-term survival of the grafted limbs, rejection rate, and functional recovery. The procedure was performed in 16 rats across a strong Brown Norway to Fischer 344 histocompatibility barrier. Eight animals served as a control group that was not given any antirejection therapy and rejected their limb within a few days. The remaining eight animals were administered a 2-week course of immunosuppressive therapy including tacrolimus (TRL; 2 mg/kg/d), mycophenolate mofetil (MMF; 15 mg/kg/d), and prednisolone (Pred; 0.5 mg/kg/d). At 2 weeks, Pred and MMF were simultaneously tapered by 20% of the dosage every week; by week 7 the animals were on TRL only. TRL was then tapered at the same rate (20% every week) to a maintenance dose of 0.6 mg/kg/d at week 12. After 6 months the immunosuppression was stopped. Four of 8 animals did not reject throughout the study up, to the 1-year endpoint. At this stage they show excellent functional outcomes, evaluated by clinical tests and walking tract analysis. The remaining four rats developed a rejection at an average of 267 days postoperatively (range 224 to 302 days), corresponding to an average of 87 days (range 44 to 122 days) without any immunosuppression. They were sacrificed as soon as rejection was confirmed for histological examination of the various tissues. This study showed that a triple combination therapy provides excellent long-term functional outcomes of the transplanted limbs, with no rejection episodes, no side effects, or complications, even 6 months after withdrawal of all immunosuppressive components, suggesting the possible emergence of tolerance. PMID- 15110629 TI - The risk of contracting an infectious disease from blood transfusion. AB - In the general population, the likelihood of an individual receiving a transfusion has been calculated to be about 0.89% per year, increasing dramatically with age. Massive intraoperative hemorrhage from trauma, cardiopulmonary bypass, and orthotopic liver transplantation need substantial replacement therapy. In renal transplantation, blood transfusion is a debated induction tool for specific allograft tolerance, since it causes a nonspecific down-regulation of immune function. In transplantations, in humoral immune deficiencies, in hematological disorders, and in HIV infection, the intravenous immunoglobulin prophylaxis may alter the monocyte/macrophage system host immunity and immune surveillance against infection, tissue or cell damage, and malignancy. Some persons, like Jehovah's Witnesses, object to transfusion of blood products, posing ethical and practical issues concerning treatment of blood disorders, transplantation, and trauma. In this review we examined the actual risk of contracting an infectious disease from an allogeneic blood transfusion to contribute to an uneasy decision-making process. We have found that the procedure is presently considerably safe. PMID- 15110630 TI - Cyclosporine monitoring in stable, long-term, pediatric kidney transplant recipients: the value of C2 determination. AB - Although a generalized consensus has been reached for therapeutic drug monitoring of cyclosporine microemulsion in adult transplant patients, clear guidelines are recently not available for the pediatric population. In this retrospective analysis of pharmacokinetic data obtained from stable, long-term, pediatric kidney transplant recipients, we sought to define a possible approach to manage cyclosporine therapy in a pediatric setting. The 2-hour postdose cyclosporine blood concentration, C(2), rather than trough levels, was the best single time point predictor of the area under the concentration curve. We concluded that therapeutic drug monitoring of cyclosporine-based immunosuppressive regimens should be tailored based on C(2) determinations for pediatric kidney transplant recipients. PMID- 15110631 TI - Prospects for personalized immunosuppression: pharmacologic tools--a review. AB - In the last few years, novel immunosuppressive agents and new formulations, including sirolimus, mycophenolic acid (the active metabolite of mycophenolate mofetil), tacrolimus, and microemulsion cyclosporine, have significantly improved the clinical outcome of transplant recipients. However, the majority of immunosuppressive agents need a constant monitoring of drug levels to reduce the risk of graft rejection as well as drug-induced toxicities. Many factors may affect the pharmacokinetic characteristics of immunosuppressive agents, potentially reducing treatment effectiveness. Absorption and metabolism of immunosuppressive drugs are influenced by patient genotype and comedications, while comorbidities (ie, diabetes and cystic fibrosis) are responsible for altered pharmacokinetics. Dose individualization in transplant recipients is performed according to their health status, graft function, and drug therapeutic range. With respect to the last issue, therapeutic drug monitoring (TDM) plays a crucial role in achieving optimal immunosuppression, improving the efficacy of drugs, and lowering toxic effects. Pharmacokinetic analysis allowed the identification of specific parameters, such as plasma or blood levels, immediately before dosing (C(min) or trough levels) or 2 hours after administration (C(2)), which are significantly related to tissue exposure to the drug. More recently, studies have investigated treatment individualization by evaluating drug pharmacogenetics based on the expression level or mutations of their molecular targets, including calcineurin for cyclosporine and tacrolimus, and inosine monophosphate dehydrogenase for mycophenolic acid. Although no conclusive data may be drawn from these preliminary trials, further studies are underway to address the role of pharmacogenetics in clinical decision making. PMID- 15110632 TI - Posttransplant diabetes mellitus after kidney transplantation with different immunosuppressive agents. AB - Posttransplant diabetes mellitus (PTDM) is a disturbing side effect of immunosuppression. The aim of this study was to evaluate the effect of different immunosuppressive agents on the development of PTDM in renal transplant recipients (KTx). The incidence of PTDM was evaluated in 538 consecutive KTx. Baseline immunosuppression was azathioprine (AZA), cyclosporine (CSA), or tacrolimus (TAC), or sirolimus in combination with calcineurin inhibitors (SIR). All patients received steroids for both induction and maintenance therapy during the first 6 months posttransplantation. Mean follow-up after KTx was 73 +/- 53.5 months (range 6 months to 16 years). PTDM was defined as two consecutive blood glucose determinations above 126 mg/dL. Thirty-six of 538 (6.7%) recipients experienced PTDM, 31 of whom required insulin treatment and five oral antidiabetic medications. PTDM occurred at 25.3 +/- 38 months posttransplantation in 4.8% of KTx treated with AZA, 4.8% of CSA, 6.5% of KTx treated with TAC and 12.5% of KTx treated with SIR. The time of onset of PTDM was significantly shorter (P =.003) among TAC (2.1 +/- 1.7 months posttransplantation) versus CSA (27.8 +/- 34 months). PTDM disappeared in 6 of 36 patients. We conclude that with current levels of immunosuppression, there is no difference in the incidence of PTDM between TAC- and CSA-treated KTx. PMID- 15110633 TI - Renal allograft protection with early angiotensin-converting enzyme inhibitors administration. AB - Twenty renal transplant recipients (RTx) with a normal ultrasound pattern of renal artery who began angiotensin-converting enzyme inhibitor (ACEI) therapy within 14 months after surgery (ACEI(+)) were studied retrospectively to evaluate endogenous creatinine clearance/1.73 m(2) body surface area (CrCl), proteinuria (UP), UP/CrCl (FUP), mean arterial pressure (MBP), total cholesterol, LDL, HDL, and triglycerides. Before (T(0)) and every month for 2 years after initiation of ACEI. Twenty-four RTx who never received ACEI (ACEI(-)) were studied in the same fashion. No differences in the parameters were noted at T(0); all RTx had CrCl >60 mL/min, Up less than 0.5 g/d, and stable renal function for 3 months before the study. In the ACEI cohort CrCl was reduced after 2 years compared with T(0) (65.6 +/- 2.8 vs 76 +/- 3.2 mL/min, P <.004), UP and FUP were both increased (660 +/- 60 vs 130 +/- 20 mg/d, 8.9 +/- 1.3 vs 2.8 +/- 0.6 mg/mL x 10(3); P <.001 and.002, respectively). UP >0.5 g/d was present in three cases. After 2 years the ACEI(+) group showed a decrease in CrCl (68.2 +/- 3.1 vs 73 +/- 2.2 mL/min) and the increase in UP (181 +/- 21 vs 139 +/- 18 mg/d) and in FUP (3.1 +/- 0.7 vs 2.6 +/- 0.9 mg/mL x 10(3)), which were not significantly different from the values at T(0). No cases showed UP >0.5 g/d. Moreover UP (P <.04), FUP (P <.03) and the percent reduction of CrCl (11.2 +/- 2.5% vs 4.6 +/- 1.8%, P <.05) were greater among ACEI(-) than ACEI(+) patients at 2 years. ACEI(-) patients showed correlation between the percent reduction of CrCl and UP (r =.51, P <.04). The values of MBP and lipids did not reveal any significant difference between the two groups. In conclusion, this study suggests that ACEI have a renoprotective effect, when used early, and may also prevent chronic allograft nephropathy. PMID- 15110634 TI - Effect of calcineurin inhibitors on extracellular matrix turnover in isolated human glomeruli. AB - INTRODUCTION: Although chronic cyclosporine toxicity is mainly characterized by tubular atrophy and interstitial fibrosis, glomerular injury with expansion of mesangial matrix and sclerosis is not uncommon. Tacrolimus is a newer calcineurin inhibitor that has been used in renal transplant recipients as primary or rescue therapy. Clinical trials suggest an improved long-term graft survival among patients treated with tacrolimus. Recently we have shown that tacrolimus and cyclosporine have similar effects on extracellular matrix turnover in cultured cells. The present study was performed to investigate the effects of the calcineurin inhibitors on whole glomeruli extracellular matrix turnover. METHODS: Human glomeruli isolated from kidney biopsies just before transplantation were incubated with culture media containing either cyclosporine (200 ng/mL) or tacrolimus (10 ng/mL) for 24 hours. Glomeruli incubated only with culture medium were used as control. RESULTS: The expressions of (alpha2)IV collagen, metalloprotease 9 (MMP9), tissue inhibitors of metalloproteases 2 (TIMP-2), and TGFbeta were evaluated by in situ reverse transcription and polymerase chain reactions (RT-PCR). beta-actin was used as a control gene. Cyclosporine (but not tacrolimus) increased the expression of (alpha2)IV collagen and TIMP2 in isolated glomeruli. TGF-beta was markedly increased by cyclosporine. MMP9 expression was not affected by the calcineurin inhibitors. By light microscopy kidney biopsies did not show pathologic changes. CONCLUSION: Cyclosporine treatment modulates extracellular matrix turnover in isolated human glomeruli, inducing an imbalance between synthesis and degradation. This effect, not observed in tacrolimus treated human glomeruli, may induce the extracellular matrix deposition and sclerosis characteristic of chronic cyclosporine toxicity. PMID- 15110635 TI - Plasma levels of transforming growth factor-beta1 in renal transplant recipients receiving different immunosuppressive regimens. AB - Experimental and clinical evidence support the role of transforming growth factor beta-1 (TGF-beta(1)), a cytokine with complex immune and nonimmune effects, on the development of chronic renal allograft nephropathy (CAN). We investigated the effects of different immunosuppressive regimens on circulating TGF-beta(1) plasma levels in stable kidney transplant (KTx) recipients. Two hundred ninety-nine TGF beta(1) plasma levels were measured in 125 kidney transplant (KTX) recipients exhibiting stable renal function, immunosuppressed with cyclosporine (CsA), tacrolimus (TAC), or sirolimus (SIR), and in 18 normal healthy volunteers (C). Activated immunoreactive TGF-beta(1) was detected in platelet-depleted plasma by an enzyme-linked immunoadsorbent assay. Multivariate analyses correlated immunosuppressive regimens with TGF-beta(1) levels. KTX recipients displayed significantly higher TGF-beta(1) levels compared to C (P =.0005). Patients receiving CsA had significantly higher TGF-beta(1) plasma levels compared to those receiving TAC or SIR (P =.0384). Multivariate analyses showed no correlation between TGF-beta(1) levels and immunosuppressive drug trough blood levels or doses, but only correlations with the main immunosuppressive drug. These data show that: (1) TGF-beta(1) production is activated in kidney transplant recipients; (2) CsA patients display significantly higher plasma TGF beta(1) levels. Follow-up studies seek to assess the possible relationship with clinical events. PMID- 15110636 TI - Platelet-Independent defect in hemostasis associated with sirolimus use. AB - Sirolimus is currently used to prevent rejection of solid organ transplant, and sirolimus-eluting stents have shown promise for the prevention of coronary artery restenosis. Thrombocytopenia is a well-known adverse effect of sirolimus limiting its use. Herein we report on a patient in whom sirolimus caused a platelet independent hemostasis defect. The patient was a 52-year-old woman who underwent renal transplant with consequent normal kidney function. The immunosuppressive regimen included basiliximab, steroids, and cyclosporine induction later shifted to sirolimus and mycophenolate due to biopsy findings of tubular necrosis on day 6 posttransplantation. At discharge the serum creatinine was 0.7 mg/dL. Four months after transplantation the patient was admitted to our hospital because of fever (37.5 degrees C to 38 degrees C), anorexia, and asthenia. Blood analysis showed: creatinine 1.7 mg/dL, Hb 9.6 g/dL, WBC 6 x 10(3)/microL, PLT 123 x 10(3)/microL, liver function tests normal, LDH 720 mU/mL, fibrinogen 628 mg/dL, d dimer 0.42 ng/mL, FDP > 40 ng/mL, INR 1.10, PT 87%, aPTT 40 seconds. Cultures and tests for infection were negative. Serum sirolimus level was 25.9 ng/mL. The following day the serum creatinine rose to 2.3 mg/dL and diuresis fell to 20 mL/h. Multiple bleeding times (Ivy test) performed before the renal biopsy were repeatedly over 30 minutes (normal 3 to 5 minutes), despite normal platelet count and platelet function studies. There was no spontaneous aggregation and in vitro aggregation was normal (collagen, ADP, adrenalin, and ristocetin induced). Coagulation studies showed a defect in fibrin formation and a reduction of fibrinolysis. Suspension of sirolimus treatment was followed by remission of fever, improvement of renal function (serum creatinine 1.2 mg/dL), and normalization of bleeding time. PMID- 15110637 TI - Hematologic toxicity of immunosuppressive treatment. AB - The administration of immunosuppressive agents may be associated with the occurrence of hematologic toxicity, such as anemia, due to bone marrow suppression or hemolysis, leukopenia, and thrombocytopenia. The administration of azathioprine and mycophenolate mofetil is more frequently associated with bone marrow suppression, while hemolytic-uremic syndrome may occur after administration of cyclosporine, tacrolimus, or muromonab (OKT3) and may be associated with the loss of the allograft. Moreover, microangiopathic hemolytic anemia and thrombocytopenia are rare, but potentially severe, complications of immunosuppressive treatment with tacrolimus and cyclosporine; they are characterized by intravascular hemolysis due to mechanical destruction of red cells as a result of pathological changes in small blood vessels. Viral infections (cytomegalovirus), administration of antiviral agents (gancyclovir), inhibitors of angiotensin-converting enzyme and angiotensin II receptor antagonists, antibacterial agents (sulfamethoxazole and trimethoprim), and allopurinol may aggravate bone marrow suppression, particularly when administered with agents that interfere with purine biosynthesis, including azathioprine and mycophenolate mofetil. PMID- 15110638 TI - Teratogenesis and immunosuppressive treatment. AB - Despite the potential risks to the mother and fetus caused by immunosuppressive drugs, uneventful pregnancies are now frequent among transplant recipients. Although there is no apparent increase in the type or incidence of malformations in the newborns or evidence of graft dysfunction, pregnancy-related complications, including premature termination and low birth weight, may be more frequent. To prevent graft rejection due to the increased immunologic reactivity of the transplant recipient during pregnancy, it is reasonable to wait 2 years after transplantation before conception, to have stable graft function and to be on low drug doses for maintenance immunosuppression. Among the immunosuppressive agents, corticosteroids may induce a number of treatment-related complications, including diabetes and osteoporosis; however, the incidence of fetal malformations during corticosteroid treatment is about 3.5%, a value close to that of the general population. Among immunosuppressive antibodies, no evidence of developmental toxicity has been demonstrated with basiliximab. On the contrary, some concerns have been raised about azathioprine, since its use has been associated with fetal abnormalities in animals; however, clinical data so far have indicated only a small teratogenic risk. Therefore, immunosuppressive therapy with selected drugs and antibodies does not apparently increase the risk of birth defects and may be continued in pregnancy. Finally, although breast feeding is not recommended, because of drug transfer into maternal milk, the available clinical data do not support this limitation because of the low amount of drug absorbed by the infant and the absence of clinical toxicity in published case reports. PMID- 15110639 TI - Tacrolimus-associated myositis: a case report in a renal transplant patient. AB - A 55-year-old Caucasian man who had received a second kidney graft in July 1993, was switched from cyclosporine to tacrolimus in June 2000 due to deterioration of renal function. Thereafter, he began to complain of muscle cramps in both quadriceps with an increased CPK and EMG findings of polyneuropathy. A muscle biopsy demonstrated acute myositis. Prednisone was administered with amelioration of the patient's symptoms, but with persistently increased CPK and myoglobin levels. In February 2001, mycophenolate mofetil was introduced and tacrolimus tapered to 3 mg daily to seek a toxic role of this immunosuppressant, since there was no other cause of myositis. A sudden decrease in CPK was observed, but the complete normalization took place only after its withdrawal in September 2002. This case represents a tacrolimus-associated myositis. PMID- 15110640 TI - Severe rhabdomyolysis and acute renal failure in a kidney transplant patient treated with tacrolimus and chimaeric CD25 monoclonal antibody. AB - Recently observations of rhabdomyolysis in patients treated with tacrolimus have been reported. The authors present a kidney transplant patient who had an epileptic seizures, severe rhabdomyolysis, and acute renal failure. The patient was initially immunosuppressed with tacrolimus and chimeric CD25 monoclonal antibody. After intensive therapy with plasmapheresis, CVVH, and dialysis, the patient completely recovered at 11/2 year his serum creatinine is 1.2 mg/dL. PMID- 15110641 TI - Polyoma virus BK and renal dysfunction in a transplanted population. AB - INTRODUCTION: Reactivation of polyoma virus BK (BKV) is increasingly recognized as a cause of severe renal-allograft dysfunction. The aim of the present study was to evaluate prevalence of BKV infection and activity in a population of kidney (KT) and liver (LT) transplant patients and search for a possible correlation with renal dysfunction. METHODS: We studied 118 patients for BKV viruria and, when present, for BKV viremia. We also assessed HCV status. RESULTS: Among 16 patients with BKV viruria (5 LT and 11 KT), eight showed BKV viremia (one LT and seven KT). Among BKV viruria-positive patients, three LT recipients were HCV-positive. All LT BKV viruria-positive patients showed normal renal function with a mean serum creatinine (sCr) blood level of 0.9 mg% and a mean blood urea nitrogen (BUN) value of about 36 mg%. The mean transplant age was 2.5 years. In contrast, KT BKV viruria-positive patients showed impaired renal function which was slightly worse in patients who also displayed BKV viremia, namely, a mean sCr blood level 1.7 mg% and a mean BUN value about 80 mg%. The mean transplant age was 7 years. CONCLUSION: Based on these findings, it seems that BKV viruria in renal allograft recipients may be associated with viremia and related to nephropathy that may lead to allograft rejection. The study will be completed with a 2-year follow-up of positive patients to assess the possible relationship between BKV active infection and eventual decrease of renal function and loss of transplanted organ. PMID- 15110642 TI - Plasma exchange for polyradiculoneuropathy following kidney transplantation: a case report. AB - We describe a case of polyradiculoneuropathy (PRN) following living donor kidney transplantation, without clinical evidence of preexisting infection. In this study plasma exchange treatment resulted 6 days later in improvement in extremity weakness and paresthesias in the upper and lower extremities. Total neurological recovery was obtained 3 months after the onset of symptoms. PMID- 15110643 TI - Early de novo malignancies after kidney transplantation. AB - INTRODUCTION: Immunosuppressed renal transplant patients display a higher incidence of carcinoma than the general population. The chronic use of immunosuppressive therapy to prevent acute rejection increases the long-term risk of cancer. We reviewed our experience to identify factors affecting the development of de novo neoplasms. PATIENTS AND METHODS: Between January 2000 and May 2003, 135 renal and three combined kidney-pancreas transplantations were performed. RESULTS: Sixteen (11.6%) cancers were diagnosed in nine renal transplant recipients (6.5%). Tumors presented at a mean time of 14 months. Three patients displayed in malignancies; three, Kaposi's sarcoma; one, papillary microcarcinoma of the thyroid; one, bladder carcinoma; and one, breast carcinoma. CONCLUSION: Although de novo malignancies occur more frequently many years after kidney transplantation, our experience demonstrates that they can occur early during the posttransplant follow-up. Skin malignancies showed the best prognosis, probably because of early detection and treatment. Patients with Kaposi's sarcoma benefit from reduction or cessation of immunosuppression, but this entails a higher risk of graft loss. Solid organ de novo malignancies are often more aggressive than those in normal population; the life expectancy of these recipients is low. PMID- 15110644 TI - Importance of biopsy evaluation and the role of the pathologist in solid organ transplant programs. PMID- 15110645 TI - Molecular techniques in transplantation. AB - Molecular tools may prove useful for diagnosis, monitoring, and definition of basic mechanisms in transplantation. These techniques must be quantitative, reproducible and rapid, displaying known sensitivity and specificity. Powerful new techniques with "real-time" application, as briefly reviewed herein, are moving closer to clinical utility. PMID- 15110646 TI - Correlation of morphological findings with functional reserve in the aging donor: role of the poly (ADP-ribose) polymerase. AB - INTRODUCTION: The enzyme poly (ADP-ribose) polymerase (PARP-1) participates in the first events of DNA repair in higher organisms. Under conditions of tissue ischemia, this action can lead to significant decreases in NAD(+), massive adenosine triphosphate (ATP) depletion, and cell death. In renal grafts with pretransplantation cold ischemia and subsequent ischemia-reperfusion injury, overactivation of PARP-1 may lead to a higher index of acute tubular necrosis, a delay in total recovery of the function of the transplanted organ, and an early progression to chronic graft nephropathy. The present study examined whether increased tubular expression of PARP-1 in kidneys from aged donors contributed to recipient renal function. MATERIAL AND METHOD: We studied the nuclear expression of PARP-1 using immunohistochemistry with monoclonal antibody PAR01 in 75 kidney biopsy specimens from 40 aged donors. RESULTS: Immunohistochemical expression of PARP-1 showed a statistically significant relationship with donor age (r =.408, P =.006, Spearman test), with time required to achieve effective diuresis (r =.386, P =.01, Spearman test) and with creatinine levels in the first 3 months. We also highlighted a greater intensity of PARP-1 expression in suboptimal donor kidneys that failed to reduce the serum creatinine levels to <1.7 mg/dL (creatinine <1.7 PARP: 1.29 +/- 1.49 vs creatinine >1.7 PARP: 2.29 +/- 1.33, P =.047, Mann-Whitney U test). CONCLUSION: We conclude that the determination of PARP-1 in biopsy specimens from aged donors may be a useful predictive factor for renal graft function. PMID- 15110647 TI - Factors that influence the development of an organ donation program. AB - The shortage of cadaveric donors is a universal problem that imposes a severe limit on the number of patients who can benefit from transplantation. At the same time, there is an ever-increasing demand for cadaveric solid organs all over the world. The organ shortage is not due to a lack of potential donors, but rather to a failure to turn many potential into actual donors. Spain is the only example in the world of continuous improvement in cadaveric organ donation registered in a large country during more than 10 years. This success is due to a proactive donor detection program performed by well-trained transplant coordinators, introduction of systematic death audits in hospitals, and the combination of a positive social atmosphere, an adequate management of mass media relations, and on adequate economic reimbursement for the hospitals. This model can be partial or totally adapted to other countries or regions, if basic conditions are guaranteed. A careful study of local characteristics, which influence organ donation in a direct or indirect way, should be performed before planning specific actions to improve organ donor rates. The principle factors that influence the development of this type of program are described in this article. PMID- 15110648 TI - Biopsy of the marginal kidney donor: correlation of histology with outcome. PMID- 15110649 TI - Influence of the management of asystolic donors on kidney transplantation outcome. AB - Two methods of donor management were analysed, namely, with and without in situ cooling perfusion of the kidney in an attempt to determine the optimal management and preservation methods for asystolic kidney donors. The group of recipients of in situ cooling perfusion kidneys showed more days of oliguria (P <.05), needed more dialysis sessions (P <.05), and showed no transplant function during the first week after surgery. This group also had a greater probability of acute rejection (P =.071) and a higher rate of nonfunctioning grafts (P =.09). We conclude that in situ cooling perfusion of asystolic kidney donors impairs graft function. PMID- 15110651 TI - The Banff schema and differential diagnosis of allograft dysfunction. AB - The pathological findings on renal allograft biopsy specimens are often complex. By carefully defining the morphological features of acute allograft rejection, the Banff schema provides an important tool for the differential diagnosis of acute allograft dysfunction. This article reviews the criteria for diagnosis of rejection, and its differentiation from other inflammatory infiltrates in renal allograft biopsy specimens. PMID- 15110650 TI - Liver transplantation from maastricht category 2 non-heart-beating donors: a source to increase the donor pool? AB - INTRODUCTION: The demand for liver transplantation has increasingly exceeded the supply of cadaver donor organs. Non-heart-beating donors (NHBDs) may be an alternative to increase the cadaver donor pool. The outcome of 20 liver transplants from Maastricht category 2 NHBD was compared with that of 40 liver transplants from heart-beating donors (HBDs). After unsuccessful cardiopulmonary resuscitation (CPR), cardiopulmonary support with simultaneous application of chest and abdominal compression (CPS; n = 6) or cardiopulmonary bypass (CPB; n = 14) was used to maintain the donors. RESULTS: At a minimum follow-up of 2 years, actuarial patient and graft survival rates with livers from Maastricht category 2 NHBD were 80% and 55%, respectively. Transplantation of organs from these donors was associated with a significantly higher incidence of primary nonfunction, biliary complications, and more severe initial liver dysfunction compared with organs from HBDs. The graft survival rates was 83% for livers from NHBDs preserved with CPS and 42% in those maintained with CPB. PMID- 15110652 TI - Borderline changes in the Banff schema: rejection or no rejection? AB - The relationship between acute renal allograft rejection and histopathologic biopsy alterations recognized by the Banff Schema as "borderline changes" is not clear. Some evidence supports the contention that about one third of patients with borderline infiltrates and clinical evidence of graft dysfunction do indeed have acute rejection, which, if left untreated, progresses to a histologically more advanced stage of rejection. Several investigators recognize that not all patients with mild tubulitis respond clinically to antirejection therapy; a significant number of these biopsy specimens display additional histological alterations. The most common concurrent lesions are chronic allograft nephropathy, arteriolar lesions consistent with calcineurin inhibitor toxicity, acute tubular necrosis, and obstructive nephropathy. Management of patients with borderline changes must tightly correlate the pathologic features and the clinical information. PMID- 15110653 TI - Improved outcome of polyoma virus allograft nephropathy with early biopsy. AB - Polyoma virus allograft nephropathy often results in accelerated graft loss despite reduction of immunosuppression and/or treatment with antiviral agents. Irreversible renal fibrosis due to late diagnosis is likely to be one of the important causes of treatment failure. Early biopsy in 14 patients resulted in stable graft function after a mean follow-up of 22 months. PMID- 15110654 TI - Hepatitis C virus infection and renal disease after renal transplantation. AB - Hepatitis C virus (HCV) infection is the main cause of chronic liver disease after renal transplantation (RT). It is considered in some series to be a risk factor for graft loss and patient death. Also, HCV has been implicated in the pathogenesis of glomerular diseases in native and transplanted kidneys. The presence of membranoproliferative (MP) or membranous (M) glomerulonephritis (GN) in HCV-positive patients has been well documented after RT, but there is no clear data concerning the real prevalence of HCV-induced glomerulonephritis. MPGN with or without cryoglobulinemia and MGN have been described in HCV RNA-positive patients in general without severe liver disease. Also, there is a possible association between HCV infection and acute/chronic transplant glomerulopathy. Renal thrombotic microangiopathy has been described in HCV-positive patients with positive anti-cardolipin antibodies. The pathogenesis of MPGN and MGN in HCV patients after RT seems to be similar to that which occurs in native kidneys: the deposition of immune complexes containing HCV proteins in the glomeruli. Renal biopsy, using light microscopy, immunofluorescence techniques, and electron microscopy, is useful to achieve a correct diagnosis. Unfortunately, interferon is not recommended due to the significant risk of rejection. The possibility of pegylated interferon needs to be tested. Ribavirin can improve proteinuria but HCV RNA remains positive. Finally, recent data suggest that the use of interferon in HCV patients on dialysis can negate HCV RNA and prevent associated glomerulonephritis after RT. PMID- 15110655 TI - Early diagnosis of chronic allograft nephropathy by means of protocol biopsies. AB - Chronic allograft nephropathy (CAN) is the first cause of graft failure. Since graft survival has improved and the incidence of rejection decreased, these outcomes cannot be employed as primary efficacy variables in clinical trials due to the need for a large sample size. The presence of CAN in protocol biopsies is an independent predictor of graft survival. Thus, it has been proposed that chronic lesions in protocol biopsies be considered a primary efficacy variable. Power calculations have confirmed this hypothesis, especially if CAN is evaluated using a morphometric technique. Moreover, it has been demonstrated that in vivo glomerular number (Ng) can be estimated by combining a protocol biopsy with magnetic resonance imaging. Ng correlates with graft function in stable grafts. Taken together, these data suggest that protocol biopsies constitute a fundamental tool to improve the design of clinical trials and to define parameters that are crucial to the understanding of mechanisms leading to CAN. PMID- 15110656 TI - Chronic allograft nephropathy: causes of death and mortality risk factors-a review of the last decade in Spain. AB - BACKGROUND: Improved immunosuppressive regimens and management strategies in renal transplantation (RT) have increased patient and graft survival during the last years. The aim of our study was to analyze the causes, risk factors, and evolution of mortality after renal transplantation. METHODS: We studied 3365 renal transplant recipients in adults (>18 years) who survived at least 1 year after transplantation in Spain during 1990, 1994, and 1998. The mortality rates and risk factors were analyzed employing single and multivariate Cox regression. RESULTS: The follow-up was shortest (maximum 2.5 years) for recipients transplanted in 1998. When we consider an identical follow-up period (2.5 years) for all patients, we did not observe a statistical difference in patient survival and causes of death in the three analyzed periods. Mortality was higher for men and for patients over 60 years. Cardiovascular diseases (CVD) and neoplasia were the most frequent causes of death. Graft dysfunction, as determined by creatinine level or proteinuria range in the first months, were significant factors associated with a higher risk for cardiovascular and infectious deaths. CONCLUSIONS: During the last decade in Spain, patient survival after RT (2.5 years follow-up) has remained stable. Recipient age (>60 years), male gender, and graft dysfunction in the first year were associated with a higher risk of death especially due to CVD. PMID- 15110657 TI - Antibody-mediated rejection in the kidney. AB - Antibody-mediated rejection is increasingly recognized as a cause of acute and chronic rejection in kidney as well as other organ allografts. New diagnostic criteria have been drafted to diagnose this entity and enable timely therapy. PMID- 15110658 TI - Histological features with clinical impact in chronic allograft nephropathy: review of 66 cases. AB - The histological features of chronic allograft nephropathy (CAN) are variable, since it is related to multiple donor and recipient factors. The main histological parameters in CAN are interstitial fibrosis, tubular atrophy, chronic vasculopathy and glomerulosclerosis. There have been many attempts to relate chronic deterioration of renal function with histologic features. MATERIALS AND METHODS: We reviewed 66 kidney transplant patients (43 men/23 women) with renal failure 6 months after transplant. The clinical data included donor and recipient age, cold ischemia time, delayed graft function (DGF), creatinine clearance, proteinuria, HLA compatibility, CMV infection, cholesterol levels, diastolic and systolic blood pressure. Banff criteria were used to grade histological parameters. The relation between clinical and histological data were analyzed using chi square, Student t, Mann Whitney and Kruskal-Wallis tests as appropriate. The cumulative graft and patient survival rates were calculated by the Kaplan-Meier method. RESULTS: The survival rate of patients with creatinine clearances >3 mg/dL at the time of the biopsy was worse than that of patients with creatinine <3 mg/dL (P =.001; log rank 20.1). We found an association between the grade of arteriosclerosis and the diastolic blood pressure (P =.017). The creatinine level was greater among patients with tabulitis than those without tubulitis (P =.06). In addition to our results we review the literature especially related to the histological feature of CAN in an attempt to detect histological findings predictive of the long term outcome of kidney allografts. PMID- 15110659 TI - Retransplantation for hepatitis C-related cirrhosis under long-term pegylated interferon therapy. AB - INTRODUCTION: Because of an increased organ shortage, one of the most controversial questions is whether hepatic retransplantation should be offered to transplant recipients with hepatitis C virus (HCV)-related graft failure because of their worse survival and the inevitable denial of other patients to access to primary transplantation. The objective of the present study was to review our experience with HCV-infected transplant recipients undergoing re-orthotopic liver transplantation (OLT) for HCV graft cirrhosis and receiving pegylated interferon and ribavirin on a prophylactic basis. RESULTS: With a median follow-up of 26 months, all 5 patients are alive with stable graft function. Four patients are still receiving pegylated interferon at a mean duration of 20 months (range, 15 32 months). Although none of the patients has cleared HCV RNA by polymerase chain reaction the mean serum levels have decreased significantly when compared with pre-retransplantation amounts. One year after re-OLT, both grade and fibrosis stage had significantly decreased; the rate of post-retransplantation fibrosis progression was significantly lower than that pre-retransplantation (3.4 +/- 0.2 vs 0.6 +/- 0.3; P <.05). PMID- 15110660 TI - Severe cardiac allograft dysfunction without endomyocardial biopsy signs of cellular rejection: incidence and management. AB - Acute dysfunction of cardiac allograft without evidence of cellular rejection is a potentially fatal complication of heart transplantation that suggests a humoral origin. In clinical practice, humoral rejection (HR) is suspected when there is evidence of severe allograft dysfunction but endomyocardial biopsy (EMB) shows no evidence of cellular rejection. Between April 1991 and August 2003, 12 patients (2.74%) among 438 heart transplants displayed this condition. Time post-heart transplant (HT) was 21.3 +/- 24.7 months (range 2 to 72 months). Immunofluorescence studies using classic markers were negative. All patients were treated with methylprednisolone "bolus" and plasmapheresis until clinical recovery, after which their immunosuppressive regimens were modified. Eleven of the 12 patients recovered satisfactory allograft function. In this series the incidence of suspected HR was low. Unlike other studies, we observed HR not only soon but also even years after HT. Plasmapheresis seems to be an effective treatment. PMID- 15110661 TI - Histological findings in "incidental" intraoperative pancreas allograft biopsies. AB - Thirty intraoperative needle core biopsies of well-functioning pancreas allografts were performed from 2 days to 7 years posttransplantation (mean 15.4 months). Most samples (83.3%) lacked significant inflammation or fibrosis. The five patients who showed features of ongoing low-grade acute rejection experienced premature graft losses due to chronic rejection. There were no complications related to the intraoperative biopsy itself. PMID- 15110662 TI - Results of pancreas transplantation in the Juan Canalejo Medical Center. PMID- 15110663 TI - Neoplasia in solid organ transplant recipients: single-center experience. AB - The incidence of neoplasms among transplanted patients is increasing, being estimated at between 4% and 18% (mean = 6%). In this article we review the neoplasms in 2514 patients who underwent transplantation in our hospital between 1981 and 2002 including 1579 kidneys, 418 hearts, 430 livers, 70 lungs, and 17 pancreas. We observed 170 tumors in 117 patients. The most frequent neoplasm was skin and lip carcinoma (30 patients) followed by PTLD (18 patients). Our results concur with the literature with one exception, a low incidence of PTLD in heart transplants, which may only be explained due to the use of preventive therapy with antiretroviral drugs. PMID- 15110664 TI - Autopsy-determined causes of death in solid organ transplant recipients. AB - The aim of this study was to review the main causes of death as determined by autopsy of deceased solid organ transplant recipients. We reviewed 156 autopsies including 76 heart, 32 liver, 29 kidney, and 19 lung transplant recipients. The mean survival period varied depending on the transplanted organ: namely heart, 497 days; liver, 189 days; kidney, 1124 days; and lung, 252 days. Infections were the most common cause of death in all groups, varying from 21% in heart to 63% in lung recipients. Acute rejection, chronic rejection, and malignancies only appeared as the cause of death in heart recipients (14.5%, 9.2%, and 4%, respectively). Primary graft failure was present in heart (15.7%), kidney (3.4%), and lung (5.3%) recipients. The highest rate of surgical complications as a cause of death was observed in heart transplant recipients. In all groups there was a significant percentage (about 30%) of other pathologies that were responsible for death, such as pulmonary embolism, central nervous system pathology, acute pancreatitis, digestive hemorrhage, and acute myocardial infarction. Our results emphasize that infections are the main cause of death within the first year posttransplant, independent of the organ transplanted. PMID- 15110665 TI - Infections following laser in situ keratomileusis: an integration of the published literature. AB - Infections occurring after laser in situ keratomileusis (LASIK) surgery are uncommon, but the number of reports have steadily increased in recent years. This systematic, comprehensive review and analysis of the published literature has been performed in order to develop an integrative perspective on these infections. We have stratified the data by potential associations, microbiology, treatment, and the degree of visual loss, using Fisher's exact tests and Student's t-tests for analysis. In this review, we found that Gram-positive bacteria and mycobacterium were the most common causative organisms. Type of postoperative antibiotic and steroid use was not associated with particular infecting organisms or severity of visual loss. Gram-positive infections were more likely to present less than 7 days after LASIK, and they were associated with pain, discharge, epithelial defects, and anterior chamber reactions. Fungal infections were associated with redness and tearing on presentation. Mycobacterial infections were more likely to present 10 or more days after LASIK surgery. Moderate or severe visual reductions in visual acuity occurred in 49.4% of eyes. Severe reductions in visual acuity were significantly more associated with fungal infections. Flap lift and repositioning preformed within 3 days of symptom onset may be associated with better visual outcome. PMID- 15110666 TI - Orbital cysts of childhood--classification, clinical features, and management. AB - There is little in the literature about the clinical spectrum of orbital cysts of childhood and no comprehensive classification has been proposed. The authors propose a classification of orbital cysts of childhood and review their clinical features, pathology, and management. The major categories in the classification include cysts of surface epithelium, teratomatous cysts, neural cysts, secondary cysts, inflammatory cysts, and noncystic lesions with cystic component. Cysts of the surface epithelium are further divided into simple epithelial cyst (epidermal, conjunctival, respiratory, and apocrine gland), and dermoid cyst (epidermal and conjunctival). Epidermal dermoid cyst (dermoid) is by far the most common orbital cystic lesion in children, accounting for over 40% of all orbital lesions of childhood and for 89% of all orbital cystic lesions of childhood that come to biopsy or surgical removal. Neural cysts include those associated with ocular maldevelopment (congenital cystic eye and colobomatous cyst) and those associated with brain and meningeal tissue (cephalocele and optic nerve meningocele). The most important secondary cyst is mucocele that can occur in children with cystic fibrosis. Inflammatory cysts are generally due to parasitic infestations and are more common in tropical areas of the world. Noncystic lesions that can have a cystic component include adenoid cystic carcinoma, rhabdomyosarcoma, lymphangioma, and others. Each type of cyst has rather characteristic, but not pathognomonic, clinical features. Computed tomography and magnetic resonance imaging can help differentiate a cystic lesion from a solid tumor, suggest the type of cyst, and help in planning management. The pathology varies with the cells that line the cyst and with the inflammatory agent. Management varies from local excision to observation, depending on the location and type of cyst. Orbital cysts of childhood can be classified into categories, based mainly on their histopathology. The clinical, radiologic, and histopathologic features can be correlated with the classification in order to better evaluate a child with a cystic lesion in the orbit. PMID- 15110667 TI - Molecular genetic basis of inherited cataract and associated phenotypes. AB - Congenital cataract is a leading cause of visual disability in children. Inherited isolated (non-syndromic) cataract represents a significant proportion of cases and recently many causative genetic mutations have been identified. Inherited cataract is known to be clinically and genetically heterogeneous. Eleven clear-cut cataract phenotypes have been described. Cataract may be inherited as autosomal dominant, autosomal recessive, or X-linked recessive traits, and 12 loci and 15 specific genes associated with inherited isolated cataract have been identified to date; it is likely that more genes remain to be discovered. The identification of remaining genes will not only improve our understanding of the mechanism of cataract formation but will shed new light on the developmental biology and biochemistry of the lens. Furthermore, it is possible that some of these genes will be implicated in the more common age related cataract, which also has a genetic component to its etiology. PMID- 15110668 TI - Transpupillary thermotherapy in the management of circumscribed choroidal hemangioma. AB - Laser photocoagulation and various radiation therapy methods have been used in the treatment of circumscribed choroidal hemangioma that produce visual loss. Recently, transpupillary thermotherapy has been employed in the management of choroidal hemangioma. Thirty-six cases of choroidal hemangioma treated with transpupillary thermotherapy are reviewed. Twenty-eight cases were reported in the literature and 10 cases were managed by the author. Transpupillary thermotherapy has been used as a primary treatment (36 cases) or secondary treatment (2 cases) for choroidal hemangioma. The goal of transpupillary thermotherapy is to achieve resolution of exudative detachment of the fovea and improvement of vision. In all eyes the foveal detachment subsided. In 26 eyes having pretreatment visual acuity >or=0.05 (20/400), the visual acuity increased in 20 (77%) eyes and remained unchanged in 6 (23%) eyes. After transpupillary thermotherapy, 16 (42%) cases showed complete regression, 20 (53%) demonstrated partial regression, and in 2 (5%) there was no change in tumor thickness at follow-ups ranging from 2 to 44 months. Complications of transpupillary thermotherapy of choroidal hemangioma included cystoid macular edema (3 eyes), preretinal fibrosis (2 eyes), focal iris atrophy (3 eyes), and retinal vascular occlusion (1 eye). Although the follow-up in many studies is limited, transpupillary thermotherapy has not been associated with any significant complications. PMID- 15110669 TI - Primary bone tumors of the orbit. AB - Primary tumors of orbital bone constitute 0.6% to 2% of all orbital tumors. Our experience over a 24-year period in the Orbital Clinic at the University of British Columbia yielded 62 (1.9%) cases from a total of 3,340 orbital tumors. Although a heterogeneous group, primary orbital bone tumors may be classified on a clinicopathologic basis into benign fibro-osseous or cartilaginous, reactive, neoplastic and vascular disorders. Presentation is usually a gradual mass effect, with infiltration and acute hemorrhage being features of malignant and reactive lesions respectively. The two most commonly encountered entities were fibrous dysplasia and osteoma, accounting for 22 cases. Although both these conditions rarely present a diagnostic challenge, nonspecific histologic and radiologic appearances can result in poor characterization of several of the rarer lesions. Hence, close cooperation between clinician, radiologist and pathologist is essential for accurate diagnosis. PMID- 15110670 TI - Rapid, painless unilateral vision loss in a 37-year-old healthy woman. AB - A 37-year-old woman experienced painless, progressive vision loss to no light perception in the left eye over the course of 3 days. The right eye was unaffected. On examination, the only other abnormal finding was a +4 left afferent pupillary defect. She was initially diagnosed with retrobulbar optic neuritis and admitted for treatment with intravenous methylprednisolone. Neuro imaging revealed a large right anterior cerebral artery aneurysm that crossed the midline to compress the left optic nerve. The aneurysm was treated with coil embolization, which was technically successful but which did not lead to significant improvement in vision. PMID- 15110671 TI - Transsphenoidal diplopia. AB - A 46-year-old man developed a pupil-involving incomplete third cranial nerve palsy after undergoing transsphenoidal resection of a pituitary macroadenoma. A CT scan 1 week later revealed postoperative changes, with no new mass lesion, hemorrhage, or orbital fracture. The third cranial nerve palsy spontaneously improved over the ensuing week. Postoperative ocular motor palsy is an uncommon complication of transsphenoidal pituitary surgery. Neuro-imaging should be performed to exclude the presence of a new compressive lesion. PMID- 15110672 TI - Quality care and practice variation: the roles of practice guidelines and public profiles. AB - Goals of the quality-of-care initiative are to improve the structure, process, and outcome of health care. The effectiveness of methods to improve quality have been largely unverified. Most methods are costly to implement and time-consuming to perform; some threaten professional autonomy. The characteristic feature of modern medicine that fuels the debate over quality is the variation in the delivery of health care. This review examines the "variation phenomenon" in medicine and the roles that practice guidelines and physician profiling have in improving health care, in general, and for adult cataract, in particular. PMID- 15110673 TI - Otto Himmler: first manufacturer of lathe cut corneo-scleral lenses. PMID- 15110675 TI - The treatment of pterygium. PMID- 15110677 TI - Use of Nd:YAG laser capsulotomy. PMID- 15110679 TI - New advances in the production of edible plant vaccines: chloroplast expression of a tetanus vaccine antigen, TetC. AB - Vaccines are a proven method of controlling disease. However there are issues with the delivery and administration of vaccines. A particular problem is that the majority of vaccines currently used are injected, which can be unsafe if needles are reused in areas where blood-borne diseases are prevalent. Vaccines targeting the mucosal immune system avoid many of the problems associated with injections. One potential form of mucosal vaccine is based on the expression of vaccine antigens in plants. Current research in this area has focused on the expression of immunogens from the plant's nuclear genome but low expression levels generally achieved using this system have limited progress. In recent work we have used the model antigen, TetC, which confers resistance to Tetanus infection, to demonstrate the feasibility of expressing vaccine antigens at high levels in the plant chloroplast. PMID- 15110680 TI - Dietary phytoestrogens and health. AB - The interest in the potential health effects of dietary phytoestrogens has increased with the findings that hormone replacement therapy is not as safe or effective as previously thought. This review summarizes the dietary sources of the phytoestrogens; isoflavonoids, stilbenes, coumestans and lignans. It also examines 105 clinical studies related to effects of phytoestrogens on bone density, cardiovascular health, cancer prevention, cognitive ability and menopausal symptoms. PMID- 15110681 TI - Natural antimycobacterial metabolites: current status. AB - Over the years the introduction of very effective drugs has revolutionized the treatment of tuberculosis. In recent years, however, emerging multiple drug resistance has become a major threat and thus calls for an urgent search for new and effective treatments for this deadly disease. This review is complementary to earlier reviews and covers more recent reports of naturally occurring compounds, and in some cases synthetic analogs, largely from plants, fungi and marine organisms that demonstrate significant activity in the in vitro bioassays against Mycobacterium tuberculosis, and other mycobacterial species. Included also are traditional medicinal uses of specific plants when utilized to treat tuberculosis and other pulmonary diseases. PMID- 15110682 TI - Austrocolorins A1 and B1: atropisomeric 10,10'-linked dihydroanthracenones from an Australian Dermocybe sp. AB - The atropisomeric austrocolorins A(1) (7) and B(1) (8), new members of the rare tricolorin class of 10,10'-coupled dihydroanthracenones, are isolated from an indigenous Australian toadstool belonging to the subgenus Dermocybe of Cortinarius, and their structure and absolute central and axial configuration is deduced from the spectroscopic data, and confirmed by chemical degradation and chiral HPLC analysis. PMID- 15110683 TI - 1,2-Dehydroreticuline synthase, the branch point enzyme opening the morphinan biosynthetic pathway. AB - A synthase which oxidizes (S)-reticuline to 1,2-dehydroreticuline has been found to occur in seedlings of opium poppy (Papaver somniferum L.). Due to its instability, this enzyme could only be partly purified (ca. 5-fold enrichment). Partial characterization at this stage of purification showed that it does not need a redox cofactor and accepts both (S)-reticuline and (S)-norreticuline as substrates. [1-(2)H, (13)C]-(R,S)-reticuline was enzymatically converted into [1 (13)C]-dehydroreticuline, which has been identified by mass spectrometry. Release of the hydrogen atom in position C-1 of the isoquinoline alkaloid during the oxidative conversion, was exploited as a sensitive assay system for this enzyme. The enzyme has a pH optimum of 8.75, a temperature optimum of 37 degrees C and the apparent K(M) value for the substrate reticuline was shown to be 117 microM. Moreover it could be demonstrated by sucrose density gradient centrifugation that the enzyme is located in vesicles of varying size. In combination with the previously discovered strictly stereoselective and NADPH dependent 1,2 dehydroreticuline reductase the detection of this enzyme, the 1,2 dehydroreticuline synthase, provides the necessary inversion of configuration and completes the pathway from two molecules of L-tyrosine via (S)-norcoclaurine to (R)-reticuline in opium poppy involving a total number of 11 enzymes. PMID- 15110684 TI - Transcriptional activation of Igl, the gene for indole formation in Zea mays: a structure-activity study with elicitor-active N-acyl glutamines from insects. AB - The indole-3-glycerol phosphate lyase Igl is the structural gene of volatile indole biosynthesis in the tritrophic interaction in maize. The gene is activated on transcriptional level with the same kinetics and to the same level by the fatty acid-amino acid conjugates (FAC's) volicitin (17S)-(N-(17 hydroxylinolenoyl)-L-glutamine) and N-linolenoyl-L-glutamine. Both conjugates are present in the regurgitates of herbivorous caterpillars. Modifications of the fatty acid moiety of the FACs greatly reduces the elicitation of Igl and only the L-stereo-isomer of the FACs shows biological activity in the system. Volicitin treatment leads to a fast increase of AOS and AOC transcription levels and methyl jasmonate application induces Igl transcription. Hence, the induction of jasmonate biosynthesis appears to be an integral part of the elicitor mediated increase of Igl gene transcription. PMID- 15110685 TI - Biosynthetic experiments with tall plants under field conditions. 18O2 incorporation into humulone from Humulus lupulus. AB - Five segments of a large hop plant (Humulus lupulus var. Hallertauer Magnum) carrying several cones were enclosed in sealed glass vessels that were gassed with (18)O(2). After 14 days, the segments were harvested and humulone and cohumulone were analysed by NMR spectroscopy and mass spectroscopy. The oxygen atoms in position 6 of humulone and cohumulone showed 9% (18)O enrichment, respectively. It follows that the C-6 hydroxy groups were introduced by oxygenase catalysis. PMID- 15110686 TI - Hygrophorones A-G: fungicidal cyclopentenones from Hygrophorus species (Basidiomycetes). AB - Twenty new 5-(hydroxyalkyl)-2-cyclopentenone derivatives (hygrophorones) could be isolated from Hygrophorus latitabundus, H. olivaceoalbus, H. persoonii, and H. pustulatus. Their fungicidal activity was exemplarily tested. The hygrophorones have structural similarities to the antibiotic pentenomycin. Chemically, hygrophorones are 2-cyclopentenones with hydroxy or acetoxy substituents at C-4 and/or C-5. An odd-numbered 1' oxidized alkyl chain (C(11), C(13), C(15), or C(17)) is attached at C-5. In addition, from H. persoonii the new gamma butyrolactone derivative [5-(E)-2-hydroxytetradexylidene-5H-furan-2-one] could be isolated. Some hygrophorones are responsible for the color reaction of the stipes of these fungi upon treatment with potassium hydroxide solution. Structural elucidations are based on 1D ((1)H, (13)C) and 2D (COSY, NOESY, HSQC, HMBC) NMR spectroscopic analyses as well as HR-FT-ICR-MS investigations. PMID- 15110687 TI - Glucosinolate biosynthesis: demonstration and characterization of the condensing enzyme of the chain elongation cycle in Eruca sativa. AB - Glucosinolates are a group of sulfur-rich thioglucoside natural products common in the Brassicaceae and related plant families. The first phase in the formation of many glucosinolates involves the chain extension of the amino acid methionine. Additional methylene groups are inserted into the side chain of methionine by a three-step elongation cycle involving 2-oxo acid intermediates. This investigation demonstrated the first step of this chain elongation cycle in a partially-purified preparation from arugula (Eruca sativa). The 2-oxo acid derived from methionine, 4-methylthio-2-oxobutanoic acid, was shown to condense with acetyl-CoA to form 2-(2'-methylthioethyl)malate. The catalyst, designated as a 2-(omega-methylthioalkyl)malate synthase, belongs to a family of enzymes that mediate the condensation of acyl-CoAs with 2-oxo acids, including citrate synthase of the citric acid cycle, and 2-isopropylmalate synthase of leucine biosynthesis. The 2-(omega-methylthioalkyl)malate synthase studied here shares properties with other enzymes of this class, but appears chromatographically distinct and is found only in extracts of plant species producing glucosinolates from chain-elongated methionine derivatives. Although the principal glucosinolates of arugula are formed from methionine that has undergone two rounds of chain elongation to form dihomomethionine, studies with substrates and substrate analogs of different chain lengths showed that the isolated enzyme is responsible only for the condensation step of the first round of elongation. PMID- 15110688 TI - Flavonoid methylation: a novel 4'-O-methyltransferase from Catharanthus roseus, and evidence that partially methylated flavanones are substrates of four different flavonoid dioxygenases. AB - Catharanthus roseus (Madagascar periwinkle) flavonoids have a simple methylation pattern. Characteristic are B-ring 5' and 3' methylations and a methylation in the position 7 of the A-ring. The first two can be explained by a previously identified unusual O-methyltransferase (CrOMT2) that performs two sequential methylations. We used a homology based RT-PCR strategy to search for cDNAs encoding the enzyme for the A-ring 7 position. Full-length cDNAs for three proteins were characterized (CrOMT5, CrOMT6, CrOMT7). The deduced polypeptides shared 59-66% identity among each other, with CrOMT2, and with CrOMT4 (a previously characterized protein of unknown function). The five proteins formed a cluster separate from all other OMTs in a relationship tree. Analysis of the genes showed that all C. roseus OMTs had a single intron in a conserved position, and a survey of OMT genes in other plants revealed that this intron was highly conserved in evolution. The three cDNAs were cloned for expression of His-tagged recombinant proteins. CrOMT5 was insoluble, but CrOMT6 and CrOMT7 could be purified by affinity chromatography. CrOMT7 was inactive with all compounds tested. The only substrates found for CrOMT6 were 3'-O-methyl-eriodictyol (homoeriodictyol) and the corresponding flavones and flavonols. The mass spectrometric analysis showed that the enzyme was not the expected 7OMT, but a B ring 4'OMT. OMTs with this specificity had not been described before, and 3',4' dimethylated flavonoids had not been found so far in C. roseus, but they are well known from other plants. The identification of this enzyme activity raised the question whether methylation could be a part of the mechanisms channeling flavonoid biosynthesis. We investigated four purified recombinant 2-oxoglutarate dependent flavonoid dioxygenases: flavanone 3beta-hydroxylase, flavone synthase, flavonol synthase, and anthocyanidin synthase. 3'-O-Methyl-eriodictyol was a substrate for all four enzymes. The activities were only slightly lower than with the standard substrate naringenin, and in some cases much higher than with eriodictyol. Methylation in the A-ring, however, strongly reduced or abolished the activities with all four enzymes. The results suggested that B-ring 3' methylation is no hindrance for flavonoid dioxygenases. These results characterized a new type of flavonoid O-methyltransferase, and also provided new insights into the catalytic capacities of key dioxygenases in flavonoid biosynthesis. PMID- 15110689 TI - Specific accumulation and revised structures of acridone alkaloid glucosides in the tips of transformed roots of Ruta graveolens. AB - The root tips of Ruta graveolens (common rue) show strong autofluorescence of acridone alkaloids, which are characteristic secondary metabolites of this plant. To study the specific distribution and accumulation of acridone alkaloids in various root segments of Ruta graveolens, root material was harvested from genetically transformed root cultures and extracts were investigated by chromatographic techniques and HPLC-(1)H NMR spectroscopy. The cells of the elongation and differentiation zones contained acridone glucosides and large amounts of acridone alkaloids, mainly rutacridone. Gravacridondiol glucoside was identified as the dominant secondary compound of the root tips and its structure revised by means of spectroscopic methods. In addition, minor acridones, including the structurally revised gravacridontriol glucoside and unknown natural products, were found in the root tip. PMID- 15110690 TI - Biosynthesis of unusual monocyclic alkenes by the diatom Rhizosolenia setigera (Brightwell). AB - Novel, polyunsaturated monocyclic sester- and triterpenes isolated from the diatom Rhizosolenia setigera (Brightwell), are biosynthesised mainly via the mevalonate pathway. The experiments involved incubation of the alga with [1 (13)C]acetate, isolation of the alkenes by extraction and silver ion HPLC, followed by determination of the labelling pattern of one of the monocyclic triterpenes by (13)C-NMR spectroscopy. In addition, the extent of (13)C incorporation was also measured by mass spectrometry which revealed that the involvement of the mevalonate route in the biosynthesis of these cyclic compounds was less than for the co-occurring acyclic highly branched isoprenoid alkenes. PMID- 15110691 TI - Control of dehydrodiferulate cross-linking in pectins from sugar-beet tissues. AB - Pectins were extracted from roots, petioles and leaves of sugar beet, and cross linked using hydrogen peroxide and peroxidase. The effects on dehydrodiferulate formation were monitored by HPLC and TLC. Dehydrodimers were formed in different proportions to those found in vivo. There was a net loss of around 50% of the phenolic groups (monomers plus dimers) during dimerisation. Gel filtration showed that root and petiole pectin, but not leaf pectin, increased in molecular weight during cross-linking. The effects of varying the cross-linking conditions were investigated, and it was found that hydrogen peroxide concentration was the most important factor in controlling both the type and amount of dehydrodiferulate formed. PMID- 15110692 TI - Venusol from Gunnera perpensa: structural and activity studies. AB - From the aqueous extract of the dry rhizomes of Gunnera perpensa the minor components pyrogallol, succinic acid, lactic acid, and the trimethyl ether of ellagic acid glucoside were isolated. The major constituent was identified as Z venusol, a phenylpropanoid glucoside. Its structure was verified by X-ray diffraction. Tests on isolated uterine smooth muscle from rats showed that the whole extract stimulated a direct contractile response and induced a state of continuous contractility of the uterus once all additives had been removed from the organ bath. By contrast, venusol did not trigger the direct contractile response but induced the state of continuous contractility once the organ bath was flushed. PMID- 15110693 TI - Two compounds from allelopathic rice accession and their inhibitory activity on weeds and fungal pathogens. AB - A flavone (5,7,4'-trihydroxy-3',5'-dimethoxyflavone), a cyclohexenone (3 isopropyl-5-acetoxycyclohexene-2-one-1) and a liquid mixture of low polarity, containing long-chain and cyclic hydrocarbons, were isolated from leaves of allelopathic rice accession PI 312777 using column chromatography. Their structures and constituents were identified by means of HR-MS, NMR and GC/MS analyses, respectively. Bioassays showed that both the flavone and cyclohexenone significantly inhibited the growth of weeds Echinochloa crus-galli, Cyperus difformis and Cyperus iris, and the spore germination of fungal pathogens Pyricularia oryzae and Rhizoctonia solani at all tested concentrations. Moreover, the combination of the inactive mixture of low polarity and the active flavone or cyclohexenone significantly enhanced the inhibitory activities on weed growth. In addition, the two compounds and the mixture of low polarity from the leaves of PI312777 did not inhibit the rice growth at the same concentrations. It was also established that both compounds could be released into the soil, and was especially induced by E. crus-galli. The results suggest that 5,7,4'-trihydroxy 3',5'-dimethoxyflavone and 3-isopropyl- 5-acetoxycyclohexene-2-one-1 may act as allelochemicals participating in the defense of rice against weeds and pathogens. PMID- 15110694 TI - Lipoxygenase inhibiting and antioxidant oligostilbene and monoterpene galactoside from Paeonia emodi. AB - Paeoninol and paeonin C, oligostilbene and monoterpene galactoside, have been isolated from the methanolic extract of the fruits of Paeonia emodi. Their structures have been assigned on the basis of spectral analysis including 1D and 2D NMR techniques. In addition, 4-hydroxybenzoic acid 3, gallic acid 4 and methyl gallate 5 have also been reported for the first time from this species. Compounds 1 and 2 have displayed potent inhibitory potential against enzyme lipoxygenase in a concentration-dependent fashion with the IC(50) values 0.77 and 99.5 microM, along with ABTS(.+) radical quenching activity with IC(50) values of 147.5 and 498.2 microM, respectively. PMID- 15110695 TI - Diarylheptanoids from the rhizomes of Zingiber officinale. AB - Seven new diarylheptanoids, i.e., (3S,5S)-3,5-diacetoxy-1,7-bis(4-hydroxy-3 methoxyphenyl)heptane, (3R,5S)-3-acetoxy-5-hydroxy-1,7-bis(4-hydroxy-3 methoxyphenyl)heptane, (3R,5S)-3,5-dihydroxy-1-(4-hydroxy-3,5-dimethoxyphenyl)-7 (4-hydroxy-3-methoxyphenyl)heptane, (5S)-5-acetoxy-1,7-bis(4-hydroxy-3 methoxyphenyl)heptan-3-one, 5-hydroxy-1-(3,4-dihydroxy-5-methoxyphenyl)-7-(4 hydroxy-3-methoxyphenyl)heptan-3-one, 5-hydroxy-1-(4-hydroxy-3-methoxyphenyl)-7 (3,4-dihydroxy-5-methoxy-phenyl)heptan-3-one and 1,5-epoxy-3-hydroxy-1-(4-hydroxy 3,5-dimethoxyphenyl)-7-(4-hydroxy-3-methoxyphenyl)heptane were isolated from the rhizomes of Chinese ginger (Zingiber officinale Roscoe), along with 25 known compounds, i.e., 8 diarylheptanoids, 14 gingerol analogs, a diterpene and 2 steroids. Their structures were elucidated by spectroscopic and chemical methods. PMID- 15110696 TI - LC/MS/NMR analysis of isomeric divanilloylquinic acids from the root bark of Fagara zanthoxyloides Lam. AB - Gradient HPLC coupled to DAD/UV, MS/MS and NMR has been applied to the rapid structure determination of three new isomeric divanilloylquinic acids from Fagara zanthoxyloides collected in Burkina Faso: 3,4-O-divanilloylquinic acid, 3,5-O divanilloylquinic acid and 4,5-O-divanilloylquinic acid. Furthermore these new compounds named burkinabins A-C could play a useful role in sickle cell disease, as the active agents of Fagara zanthoxyloides are said to be unidentified aromatic compounds with carboxylic acid grouping (Adesanya, S.A., Sofowora, A., 1983. Biological standardisation of Zanthoxylum roots for antisickling activity. Planta Med. 48, 27-33). PMID- 15110697 TI - Sablacaurin A and B, two 19-nor-3,4-seco-lanostane-type triterpenoids from Sabal causiarum and Sabal blackburniana, respectively. AB - In search for bioactive compounds from Sabal species, sablacaurin A [25-ethyl,23 methyl-19-nor-24-methylene-3,4-seco-4(28)-lanosten-10,3-olide] and sablacaurin B [24-ethyl,24-methyl-19-nor-3,4-seco-4(28),25(26)-lanostadiene-10,3-olide], the first 19-nor lanostane derivatives of the 3,4-seco type with a spiro element, have been isolated from the leaves of Sabal causiarum and Sabal blackburniana respectively, together with the known squalene (S. blackburniana) and ss sitosterol (S. causiarum). From leaves of Sabal peregrina, the known triterpenes 3-oxo-24-methylenecycloartane and 24-methylcycloart-25(26)-en-3-one were isolated. The structures of these compounds were established from spectroscopic studies. PMID- 15110698 TI - Cytotoxic lupane-type triterpenoids from Acacia mellifera. AB - One new and eight previously described lupane-type metabolites were isolated for the first time from Acacia mellifera (Leguminosae). Based on spectral analyses, the structure of the new compound was elucidated as 28-hydroxy-3-oxo-lup-20-(29) en-30-al (1), while the known compounds were identified as 3-oxo-lup-20-(29)-en 30-al (2), 3-hydroxy-lup-20-(29)-en-30-al (3), 28-hydroxy-lup-20-(29)-en-3-one (4), lupenone (5), lupeol (6), betulin (7), betulinic acid (8), and betulonic acid (9). Metabolites 2, 3, and 4 are reported for the first time in the Leguminosae family. The cytotoxicity of the isolated metabolites was evaluated on the NSCLC-N6 cell line, derived from a human non-small-cell bronchopulmonary carcinoma. Compounds 1 and 3 exhibited significant levels of activity. PMID- 15110700 TI - ent-Kaurene diterpenoids from Isodon oresbius. AB - Three new ent-kaurene diterpenoids, oreskaurins A-C (1-3), together with ten known ent-kaurene diterpenoids, enmenin monoacetate (4), effusanin E (5), adenolin B (6), maoecrystal G (7), enmelol (8), trichokaurin (9), sodoponin (10), trichorabdal A (11), nodosin (12), enmein (13), and a flavonoid, vitexin (14), were isolated from Isodon oresbius. Their structures were determined by spectroscopic means. Compound 12 showed inhibitory activity toward K562 cells with IC(50)=1.43 microg/ml. PMID- 15110699 TI - Diterpenes from the leaves of Croton zambesicus. AB - Two new trachylobane- and one isopimarane-type diterpenoids: ent-18-hydroxy trachyloban-3-one; ent-trachyloban-3-one; isopimara-7,15-dien-3beta-ol, were isolated from the leaves of Croton zambesicus, together with trans-phytol, beta sitosterol, alpha-amyrin and stigmasterol. The structures were determined by extensive NMR techniques and X-ray analysis. The cytotoxicity of these compounds has been evaluated on cancer and non-cancer cell-lines. PMID- 15110701 TI - Thelephantins I-N; p-terphenyl derivatives from the inedible mushroom Hydnellum caeruleum. AB - Phytochemical investigation of the methanolic extract of the fruiting bodies of the inedible mushroom Hydnellum caeruleum resulted in the isolation of six p terphenyl derivatives named thelephantins I-N (1-6), together with a known compound, dihydroaurantiacin dibenzoate (7). These structures were determined by high-resolution MS, 2D NMR, IR and UV spectroscopic analysis, and by the chemical reactions. PMID- 15110702 TI - A benzil and isoflavone derivatives from Derris scandens Benth. AB - A benzil derivative: scandione, 2',2"-dihydroxy-4'-methoxy-4",5" methylenedioxybenzil and two isoflavones: scandenal, 3'-formyl-4',5-dihydroxy 2",2"-dimethylchromeno-[6,7:5",6"]isoflavone and scanderone, 4',5-dihydroxy-3' prenyl-2",2"-dimethylchromeno-[7,8:6",5"]isoflavone together with fifteen known compounds were isolated from the stem of D. scandens. Their structures were determined by spectroscopic methods. Radical scavenging, antibacterial and hypertensive activities of some of the compounds were investigated. PMID- 15110703 TI - Differential segregation and modification of mRNA during spermiogenesis in Marsilea vestita. AB - We are interested in the mechanisms that underlie cell fate determination in the endosporic male gametophytes of the fern, Marsilea vestita. Synchronous development is initiated by placing dry spores into water and involves the translation of stored mRNAs, with little transcription. Nine division cycles produce 32 spermatids surrounded by 7 sterile cells, and then each spermatid differentiates into a multiciliate gamete. Here, we focus on changes in the distribution of particular proteins, mRNAs, and patterns of polyadenylation as essential prerequisites for cell fate determination and gametogenesis. Earlier, we showed that alpha- and beta-tubulin proteins become concentrated in spermatogenous initials, and that centrin mRNA is translated only in spermatogenous initials. In situ hybridizations reveal that centrin, cyclin B, and beta-tubulin mRNAs are present in both sterile and spermatogenous cells, but that transcripts encoding RNA helicase and PRP-19 (a spliceosome component) become localized in spermatogenous cells. The targeted destruction of these two transcripts by RNAi treatments does not affect the numbers of division cycles, but the gametophytes exhibit anomalous patterns of cytokinesis, and a subsequent failure of spermatid differentiation. Thus, cell fate determination in the gametophyte involves localized translation, and the localization of mRNAs for proteins involved in transcript processing. We found differences in polyadenylation levels in sterile and spermatogenous cells that match the distribution of cytoplasmic poly(A) polymerase (PAP), which, in immunolocalizations, is abundant in spermatogenous cells, but undetectable in sterile cells. The activation of translation in spermatogenous initials, but not in sterile cells, may be under the control of mRNA processing enzymes, which become localized either as proteins or mRNAs in the spermatogenous subdomains before any divisions occur. PMID- 15110704 TI - Developmental and evolutionary aspects of the basic helix-loop-helix transcription factors Atonal-like 1 and Achaete-scute homolog 2 in the jellyfish. AB - The close functional link of nerve and muscle cells in neuromuscular units has led to the hypothesis of a common evolutionary origin of both cell types. Jellyfish are well suited to evaluate this theory since they represent the most basal extant organisms featuring both striated muscle and a nervous system. Here we describe the structure and expression of two novel genes for basic helix-loop helix (bHLH) transcription factors, the Achaete-scute B family member Ash2 and the Atonal-like gene Atl1, in the hydrozoan jellyfish Podocoryne carnea. Ash2 is expressed exclusively in larval and adult endoderm cells and may be involved in differentiation of secretory cells. Atl1 expression is more widespread and includes the developing striated muscle as well as mechanosensory and nerve cell precursors in the medusa tentacles. Moreover, Atl1 expression is upregulated in proliferating nerve cell precursors arising from adult striated muscle cells by transdifferentiation in vitro. Likewise, the neuronal marker gene NP coding for the RFamide neuropeptide is expressed not only in mature nerve cells but also transiently in the developing muscle. The molecular evidence is concurrent to the hypothesis that muscle and nerve cells are closely linked in evolution and derive from a common myoepithelial precursor. PMID- 15110705 TI - Djeyes absent (Djeya) controls prototypic planarian eye regeneration by cooperating with the transcription factor Djsix-1. AB - A conserved network of nuclear proteins is crucial to eye formation in both vertebrates and invertebrates. The finding that freshwater planarians can regenerate eyes without the contribution of Pax6 suggests that alternative combinations of regulatory elements may control the morphogenesis of the prototypic planarian eye. To further dissect the molecular events controlling eye regeneration in planarians, we investigated the role of eyes absent (Djeya) and six-1 (Djsix-1) genes in Dugesia japonica. These genes are expressed in both regenerating eyes and in differentiated photoreceptors of intact adults. Through RNAi studies, we show that Djsix-1 and Djeya are both critical for the regeneration of normal eyes in planarians and genetically cooperate in vivo to establish correct eye cell differentiation. We further demonstrate that the genetic interaction is mediated by physical interaction between the evolutionarily conserved domains of these two proteins. These data indicate that planarians use cooperatively Djsix-1 and Djeya for the proper specification of photoreceptors, implicating that the mechanism involving their evolutionarily conserved domains can be very ancient. Finally, both Djsix-1 and Djeya double stranded RNA are substantially more effective at producing no-eye phenotypes in the second round of regeneration. This is probably due to the significant plasticity of the planarian model system, based on the presence of a stable population of totipotent stem cells, which ensure the rapid cell turnover of all differentiated cell types. PMID- 15110706 TI - Differences between human and mouse embryonic stem cells. AB - We compared gene expression profiles of mouse and human ES cells by immunocytochemistry, RT-PCR, and membrane-based focused cDNA array analysis. Several markers that in concert could distinguish undifferentiated ES cells from their differentiated progeny were identified. These included known markers such as SSEA antigens, OCT3/4, SOX-2, REX-1 and TERT, as well as additional markers such as UTF-1, TRF1, TRF2, connexin43, and connexin45, FGFR-4, ABCG-2, and Glut 1. A set of negative markers that confirm the absence of differentiation was also developed. These include genes characteristic of trophoectoderm, markers of germ layers, and of more specialized progenitor cells. While the expression of many of the markers was similar in mouse and human cells, significant differences were found in the expression of vimentin, beta-III tubulin, alpha-fetoprotein, eomesodermin, HEB, ARNT, and FoxD3 as well as in the expression of the LIF receptor complex LIFR/IL6ST (gp130). Profound differences in cell cycle regulation, control of apoptosis, and cytokine expression were uncovered using focused microarrays. The profile of gene expression observed in H1 cells was similar to that of two other human ES cell lines tested (line I-6 and clonal line H9.2) and to feeder-free subclones of H1, H7, and H9, indicating that the observed differences between human and mouse ES cells were species-specific rather than arising from differences in culture conditions. PMID- 15110707 TI - Inactivation of dispatched 1 by the chameleon mutation disrupts Hedgehog signalling in the zebrafish embryo. AB - Searches of zebrafish EST and whole genome shotgun sequence databases for sequences encoding the sterol-sensing domain (SSD) protein motif identified two sets of DNA sequences with significant homology to the Drosophila dispatched gene required for release of secreted Hedgehog protein. Using morpholino antisense oligonucleotides, we found that inhibition of one of these genes, designated Disp1, results in a phenotype similar to that of the "you-type" mutants, previously implicated in signalling by Hedgehog proteins in the zebrafish embryo. Injection of disp1 mRNA into embryos homozygous for one such mutation, chameleon (con) results in rescue of the mutant phenotype. Radiation hybrid mapping localised disp1 to the same region of LG20 to which the con mutation was mapped by meiotic recombination analysis. Sequence analysis of disp1 cDNA derived from homozygous con mutant embryos revealed that both mutant alleles are associated with premature termination codons in the disp1 coding sequence. By analysing the expression of markers of specific cell types in the neural tube, pancreas and myotome of con mutant and Disp1 morphant embryos, we conclude that Disp1 activity is essential for the secretion of lipid-modified Hh proteins from midline structures. PMID- 15110708 TI - Defining the spermatogonial stem cell. AB - Through the use of donor cells from transgenic rats expressing GFP exclusively in the germline, we have defined culture conditions where male germ cells lose (on STO cells) or maintain (on MSC-1 cells) stem cell activity. A cadre of germ cell transcripts strikingly decrease in relative abundance as a function of testis age or culture time on STO cells, but only a subset of these transcripts (approximately 248) remain elevated when cultured on MSC-1 cells. If specific gene expression regulates stem cell activity, some or all of these transcripts are candidates as such regulators. We establish a spermatogonial stem cell index (SSCI) that reliably predicts relative stem cell activity in rat or mouse testis cell cultures, and through the use of an antibody to a robust signal (Egr3) within the index find intense signals in single or paired cells. As germ cells form longer interconnected chains (incomplete cytokinesis), the Egr3 signal disappears coincident with a loss of stem cell activity. Thus, molecular markers specific for spermatogonial stem cells establish a reliable and rapid means by which to define these cells in culture and alleviate the need for laborious testicular transfers in initial cell culture studies. PMID- 15110709 TI - The repressor function of snail is required for Drosophila gastrulation and is not replaceable by Escargot or Worniu. AB - Mesoderm formation in the Drosophila embryo depends on the maternal Toll signaling pathway. The Toll pathway establishes the Dorsal nuclear gradient, which regulates many zygotic genes to establish the mesodermal fate and promote the invagination of ventral cells. An important target gene of Dorsal is snail, which is required for proper mesoderm invagination. The Snail protein contains five zinc fingers and is a transcriptional repressor. However, it is not clear whether repressing target genes is a requirement for Snail to control ventral invagination. To examine such requirement, we conducted a series of genetic rescue experiments in snail mutant embryos. Snail, Worniu, and Escargot are closely related zinc-finger proteins and have equal functions during neuroblast development. However, among these three proteins, only Snail can rescue the mesoderm invagination phenotype. Moreover, the ability of various Snail mutant constructs to repress gene expression correlates with their ability to control invagination. This unique property of Snail in mesoderm formation can be attributed mostly to the CtBP co-repressor interaction motifs in the N-terminus, not to the C-terminal DNA-binding zinc fingers. Ectopic expression of Snail outside the ventral domain is not sufficient to induce cell movement even though repression of target genes still occurs. Together, the results show that the repressor function of Snail is essential for gastrulation. The repression of target genes by Snail may permit other factors in the ventral cells to positively promote mesoderm invagination. PMID- 15110710 TI - Formation and function of the polar body contractile ring in Spisula. AB - Initial studies suggested that spatial organization of the putative polar body contractile ring was determined by the peripheral aster in Spisula [Biol. Bull. 205 (2003) 192]. Here we report detailed supporting observations, including testing of aster and ring function with inhibitors. The metaphase peripheral aster was confirmed to spread cortically in an umbrella-like pattern, with microtubule-poor center. The aster disassembled during anaphase, leaving the spindle docked at the F-actin-poor center of a newly generated cortical F-actin ring that closely approximated the aster in location, measured diameter range, and pattern. Cytochalasin D and latrunculin-B permitted all events except ring and polar body formation. Nocodazole disassembly or taxol stabilization of the peripheral aster produced poorly defined rings or bulging anaphase asters within the ring center, respectively, inhibiting polar body formation. Polar body extrusion occurred at the ring center, the diameter of which diminished. Ring contractility-previously assumed-was verified using blebbistatin, a myosin-II ATPase inhibitor that permitted ring assembly but blocked polar body extrusion. The data support the hypothesis that peripheral aster spreading, perhaps dynein driven, is causally related to polar body contractile ring formation, with anaphase entry and aster disassembly also required for polar body biogenesis. Previously reported astral spreading during embryonic micromere formation suggests that related mechanisms are involved in asymmetric somatic cytokinesis. PMID- 15110711 TI - Retinoic acid and the control of dorsoventral patterning in the avian spinal cord. AB - The development of neural subtypes in the dorsoventral (DV) axis of the vertebrate central nervous system (CNS) involves the integration of signalling pathways coupled with the combinatorial expression of homeodomain transcription factors. Previous studies have implicated a role for retinoic acid in the specification of a subtype of motor neurons (MN) and in the patterning of a group of interneurons within the ventral spinal cord. In this study, we use the vitamin A-deficient (VAD) quail model to further investigate the role of retinoids in the patterning of the neural tube. Using genetic markers specific to neuronal cell populations, we demonstrate that in the absence of retinoic acid, there is a disruption to the molecular mechanisms associated with the dorsoventral patterning of the spinal cord. In particular, we observe an uneven dorsal expansion of ventral-specific genes, accompanied by a reduction in the domain of roof plate and dorsal patterning genes, both of which are rescued upon addition of retinoids during development. In addition, there is a loss of V1 interneuron specific gene expression and a decrease in the ventricular zone expression of motor neuron patterning genes. Interestingly, these effects are localised to the rostral half of the spinal cord, indicating that RA is integrated in both anteroposterior (AP) and dorsoventral patterning processes. Using differential display techniques, we have isolated 27 retinoic acid-regulated genes within the spinal cord that together reveal several interesting potential biological functions for retinoids within the avian neural tube. In summary, we propose that retinoids have an essential role in the patterning of the dorsoventral axis of the spinal cord, and are also required for the correct integration of anteroposterior patterning signals with dorsoventral determinants in the rostral spinal cord. PMID- 15110712 TI - Neurogenin3 delineates the earliest stages of spermatogenesis in the mouse testis. AB - In mammalian testis, a typical stem cell system ensures continuous spermatozoa production. Lines of experiments have demonstrated that stem cell activity resides in the most primitive small subset of germ cells, that is, A(s) (A(single)), A(pr) (A(paired)), and A(al) (A(aligned)) spermatogonia, also collectively called undifferentiated spermatogonia. However, their cellular or molecular nature is largely to be elucidated because a gene that is specifically expressed in these cells has not yet been identified, which makes it difficult to study them. In this study, we demonstrate that a class B basic helix-loop-helix (bHLH) transcription factor neurogenin3 (ngn3) is expressed specifically in A(s), A(pr), and A(al) spermatogonia because ngn3 is expressed in c-Kit negative spermatogonia throughout the seminiferous cycle, and transgenic labeling with GFP revealed connection of 1, 2, 4, 8, 16, or 32 ngn3-positive cells via intercellular bridges. ngn3 is first expressed at the prepubertal stage in c-Kit negative prespermatogonia. Lineage tracing, using the Cre-loxP system, demonstrates that ngn3-positive germ cells give rise to eventually all the spermatogenesis in mature testis. To our knowledge, ngn3 is the first reported gene that delineates these earliest stages of spermatogenesis. Considering its molecular nature, ngn3 could be involved in their differentiation control. Moreover, visualization with GFP and targeting expression of exogenous genes are valuable tools to investigate the mammalian spermatogenic stem cell system. PMID- 15110713 TI - A new culturing strategy optimises Drosophila primary cell cultures for structural and functional analyses. AB - Neurons in primary cell cultures provide important experimental possibilities complementing or substituting those in the nervous system. However, Drosophila primary cell cultures have unfortunate limitations: they lack either a range of naturally occurring cell types, or of mature physiological properties. Here, we demonstrate a strategy which supports both aspects integrated in one culture: Initial culturing in conventional serum-supplemented Schneider's medium (SM(20K)) guarantees acquisition of all properties known from 30 years of work on cell type specific differentiation in this medium. Through subsequent shift to newly developed active Schneider's medium (SM(active)), neurons adopt additional mature properties like the ability to carry out plastic morphological changes, neurotransmitter expression and electrical activity. We introduce long-term FM dye measurements as a tool for Drosophila primary cell cultures demonstrating the presence of increased, action potential-dependent synaptic activity in SM(active). This is confirmed by patch-clamp recordings, which in addition show that SM(active)-cultured neurons display different spiking patterns. Furthermore, we demonstrate that transmission can be evoked in SM(active) cultures, revealing the existence of synaptic plasticity. Thus, these culture conditions support developmental, structural and physiological properties known or expected from the nervous system, enhancing possibilities for future experiments complementing or substituting those in nervous systems of Drosophila. PMID- 15110714 TI - Conditional inactivation of Pax6 in the pancreas causes early onset of diabetes. AB - Pax6 transcription factor is required for islet cell number, morphology, and hormone gene expression. The perinatal lethality of Pax6 null mutants has restricted investigation of the role of Pax6 in normal endocrine cell function. Therefore, we devised the conditional inactivation of Pax6 using the Pdx1 and Pax6 regulatory domains to activate Cre in cells of either the entire pancreatic bud or only in endocrine cell lineages, respectively. Mutant pups died few days after birth, suffering from an overt diabetic phenotype that includes hyperglycemia, hypoinsulinemia, weight loss, and ketosis, indicating an essential role for Pax6 in beta cell function. Glucose-transporter type-2 expression was downregulated, but expression of several transcription factors essential for endocrine development was maintained. Our findings support a role for Pax6 activity in maintaining normal beta cell function after birth, but not for beta cell neogenesis during late embryonic development and early postnatal stages. PMID- 15110715 TI - A proliferative role for Wnt-3a in chick somites. AB - The proper patterning of somites to give rise to sclerotome, dermomyotome, and myotome involves the coordination of many different cellular processes, including lineage specification, cell proliferation, cell death, and differentiation, by intercellular signals. One such family of secreted signaling proteins known to influence somite patterning is the Wnt family. Although the participation of Wnt 3a in the patterning of dorsal structures in the somite is well established, no clear consensus has emerged about the cellular processes that are governed by Wnt 3a in the somite. The recent demonstration that Wnt-3a has a proliferative role in the neural tube [Development 129 (2002) 2087] suggested that Wnt-3a might also act to regulate proliferation in somites. To test this hypothesis, we first analyzed the effects of Wnt-3a on segmental plate and somite explants (from Hamburger and Hamilton stage 10 chick embryos) grown in culture. These studies indicate that Wnt-3a is capable of maintaining and/or inducing expression of both Pax-3 and Pax-7, transcription factors that have been implicated in proliferation. To directly test for a role in proliferation, explants were immunostained with antibodies against phospho-histone H3. Explants treated with Wnt-3a show an increase in the percentage of cells expressing phospho-histone H3 as compared to controls. To test the proliferative effect of Wnt-3a in vivo, we ectopically expressed Wnt-3a in chick neural tubes via electroporation. Consistent with previous studies, ectopic expression of Wnt-3a in vivo results in a mediolateral expansion of the dermomyotome and myotome. We now show that proliferation of dorsal/dermomyotomal cells is significantly enhanced by ectopic Wnt-3a. Collectively, our explant and in vivo studies indicate that an increase in proliferation plays an important role in the expansion of the dermomyotome and myotome in Wnt-3a-treated embryos. Furthermore, our results demonstrate that small changes in proliferation can dramatically influence patterning and morphogenesis. PMID- 15110716 TI - Bone morphogenetic protein-2 can mediate myocardial regulation of atrioventricular cushion mesenchymal cell formation in mice. AB - Transformation of endocardial endothelial cells into invasive mesenchyme is a critical antecedent of cardiac cushion tissue formation. The message for bone morphogenetic protein (BMP)-2 is known to be expressed in myocardial cells in a manner consistent with the segmental pattern of cushion formation [Development 109(1990) 833]. In the present work, we localized BMP-2 protein in atrioventricular (AV) myocardium in mice at embryonic day (ED) 8.5 (12 somite stage) before the onset of AV mesenchymal cell formation at ED 9.5. BMP-2 protein expression was absent from ventricular myocardium throughout the stages examined. After cellularization of the AV cushion at ED 10.5, myocardial BMP-2 protein expression was diminished in AV myocardium, whereas cushion mesenchymal cells started expressing BMP protein. Expression of BMP-2 in cushion mesenchyme persisted during later stages of development, ED 13.5-16, during valuvulogenesis. Intense expression of BMP-2 persisted in the valve tissue in adult mice. Based on the expression pattern, we performed a series of experiments to test the hypothesis that BMP-2 mediates myocardial regulation of cardiac cushion tissue formation in mice. When BMP-2 protein was added to the 16-18 somite stage (ED 9.25) AV endocardial endothelium in culture, cushion mesenchymal cells were formed in the absence of AV myocardium, which invaded into collagen gels and expressed the mesenchymal marker, smooth muscle (SM) alpha-actin; whereas the endothelial marker, PECAM-1, was lost from the invaded cells. In contrast, when noggin, a specific antagonist to BMPs, was applied together with BMP-2 to the culture medium, AV endothelial cells remained as an epithelial monolayer with little expression of SM alpha-actin, and expression of PECAM-1 was retained in the endocardial cells. When noggin was added to AV endothelial cells cocultured with associated myocardium, it blocked endothelial transformation to mesenchyme. AV endothelium treated with BMP-2 expressed elevated levels of TGFbeta-2 in the absence of myocardium, as observed in the endothelium cocultured with myocardium. BMP-2-supported elevation of TGFbeta-2 expression in endocardial cells was abolished by noggin treatment. These data indicated that BMP signaling is required in and BMP-2 is sufficient for myocardial segmental regulation of AV endocardial cushion mesenchymal cell formation in mice. PMID- 15110717 TI - Spatial pattern of nonmuscle myosin-II distribution during the development of the Drosophila compound eye and implications for retinal morphogenesis. AB - Nonmuscle myosin-II is a motor protein that drives cell movement and changes in cell shape during tissue and organ development. This study has determined the dynamic changes in myosin-II distribution during Drosophila compound eye morphogenesis. In photoreceptor neurons, myosin-II is undetectable at the apical domain throughout the first half of pupal life, at which time this membrane domain is involuted into the epithelium and progresses toward the retinal floor. Myosin-II is deployed at the apical surface at about 60% of pupal development, once the developing rhabdomeres reach the retinal floor. Subsequently, myosin-II becomes restricted to two stripes at the sides of the developing rhabdomere, adopting its final position within the visual cells R1-6; here, myosin-II is associated with a set of actin filaments that extend alongside the rhabdomeres. At the midpupal stage, myosin-II is also incorporated into stress-fiber-like arrays within the basal endfeet of the pigment cells that then change their shape. This spatiotemporal pattern of myosin-II localization and the morphological defects observed in the eyes of a myosin-II mutant suggest that the myosin-II/F-actin system is involved in the alignment of the rhabdomeres within the retina and in the flattening of the retinal floor. The observation that the myosin-II/F-actin arrays are incomplete or disorganized in R7/R8 and in rhodopsin 1-null R1-6 suggests further that the establishment and stability of this cytoskeletal system depend on rhodopsin-1 expression. PMID- 15110718 TI - An otx cis-regulatory module: a key node in the sea urchin endomesoderm gene regulatory network. AB - An essential node in the gene regulatory network (GRN) for endomesoderm specification in the sea urchin embryo lies within the regulatory system of the otx gene. According to the predictions of the GRN, based on perturbation analysis and expression data, the beta1/2 transcription unit of this gene is engaged during specification in interactions with two other regulatory genes, krox and gatae. It is predicted to be driven into activity by the krox gene, to form a positively reinforcing functional loop with the gatae gene, and to respond to its own output as well. Here we isolate the relevant otx cis-regulatory element, and examine the specific input predictions of the GRN at the level of its genomic DNA sequence. This beta1/2-otx regulatory module performs the necessary functions, as shown by use of expression constructs. It requires gatae, otx, and krox inputs, as predicted, and it operates as an "AND" logic processor in that removal of any one of these inputs essentially destroys activity. The necessary target sites were identified in the module sequence, and mutation of these sites was demonstrated to produce the same respective effects on construct expression as does blocking its regulatory inputs by treatment with morpholino antisense oligonucleotides. For spatial expression in the endoderm, one particular pair of Gata sites is essential and these function synergistically with an adjacent Otx site. We thus demonstrate directly the structure/function relationships of the genomic regulatory code, at this key node of the endomesoderm GRN. PMID- 15110719 TI - Analysis of Spemann organizer formation in Xenopus embryos by cDNA macroarrays. AB - The understanding of vertebrate development has greatly benefited from the study of gastrulation in the Xenopus embryo. Over the years, the molecular dissection of the Spemann organizer has proven to be a very fruitful source for gene discovery. Here, we report a comprehensive screen of gene expression in the Xenopus gastrula using cDNA macroarrays. Nylon filters containing more than 72000 cDNAs from a gastrula stage library were hybridized with differential probes from embryos in which organizer induction had been inhibited by reducing Nodal-related or maternal beta-Catenin signaling. Combining the changes in gene expression levels caused by these two major signaling pathways in a single graph identified both known and novel dorsoventral regulated genes. The most highly enriched organizer-specific genes were the secreted molecules chordin and Xnr-3, followed by the transmembrane protein paraxial protocadherin (PAPC). Ventral-specific abundant cDNAs included S10-40-H5, members of the Hyaluronan synthase family, Xvent-2 and XFD2/FoxI1. A differential probe of dorsal and ventral lips identified many more organizer-specific cDNAs than the screens inhibiting Nodal related and beta-Catenin signaling, suggesting that additional, as yet uncharacterized signaling pathways, contribute to organizer formation. Finally, extension of this approach to the blastula preorganizer signaling center identified the transcription factor pintallavis/FoxA2 as a new preorganizer component. PMID- 15110720 TI - Involvement of Pax6 and Otx2 in the forebrain-specific regulation of the vertebrate homeobox gene ANF/Hesx1. AB - During early vertebrate development, ANF homeobox genes are expressed in the prospective forebrain. Their regulation is essential for correct morphogenesis and function of the prosencephalon. We identified a 1-kb fragment upstream of the chicken GANF gene sufficient to drive lacZ expression in the endogenous expression domain. Concordant with the high conservation of this sequence in five investigated species, this element is also active in the corresponding expression domain of the zebrafish orthologue. In vivo analysis of two in vitro-identified Otx2 binding sites in this conserved sequence revealed their necessity for activation of the chicken ANF promoter. In addition, we identified a Pax6-binding site close to the transcriptional start site that is occupied in vivo by Pax6 protein. Pax6 and GANF exhibit mutually exclusive expression domains in the anterior embryonic region. Overexpression of Pax6 in chick embryos inhibited the endogenous GANF expression, and in Pax6(-/-) mice the expression domain of the murine ANF orthologue Hesx1 was expanded and sustained, indicating inhibitory effects of Pax6 on GANF. However, a mutation of the Pax6 site did not abolish reporter activity from an electroporated vector. We conclude that Otx2 and Pax6 are key molecules involved in conserved mechanisms of ANF gene regulation. PMID- 15110721 TI - Sox1 acts through multiple independent pathways to promote neurogenesis. AB - Although Sox1, Sox2, and Sox3 are all part of the Sox-B1 group of transcriptional regulators, only Sox1 appears to play a direct role in neural cell fate determination and differentiation. We find that overexpression of Sox1 but not Sox2 or Sox3 in cultured neural progenitor cells is sufficient to induce neuronal lineage commitment. Sox1 binds directly to the Hes1 promoter and suppresses Hes1 transcription, thus attenuating Notch signaling. Sox1 also binds to beta-catenin and suppresses beta-catenin-mediated TCF/LEF signaling, thus potentially attenuating the wnt signaling pathway. The C-terminus of Sox1 is required for both of these interactions. Sox1 also promotes exit of cells from cell cycle and up-regulates transcription of the proneural bHLH transcription factor neurogenin 1 (ngn1). These observations suggest that Sox1 works through multiple independent pathways to promote neuronal cell fate determination and differentiation. PMID- 15110722 TI - An enhancer directs differential expression of the linked Mrf4 and Myf5 myogenic regulatory genes in the mouse. AB - The myogenic regulatory factors, Mrf4 and Myf5, play a key role in skeletal muscle formation. An enhancer trap approach, devised to isolate positive-acting elements from a 200-kb YAC covering the mouse Mrf4-Myf5 locus in a C2 myoblast assay, yielded an enhancer, A17, which mapped at -8 kb 5' of Mrf4 and -17 kb 5' of Myf5. An E-box bound by complexes containing the USF transcription factor is critical for enhancer activity. In transgenic mice, A17 gave two distinct and mutually exclusive expression profiles before birth, which correspond to two phases of Mrf4 transcription. Linked to the Tk or Mrf4 minimal promoters, the nlacZ reporter was expressed either in embryonic myotomes, or later in fetal muscle, with the majority of Mrf4 lines showing embryonic expression. When linked to the Myf5 minimal promoter, only fetal muscle expression was detected. These observations identify A17 as a sequence that targets sites of myogenesis in vivo and raise questions about the mutually exclusive modes of expression and possible promoter/enhancer interactions at the Mrf4-Myf5 locus. PMID- 15110723 TI - Drawing lines in the sand: even skipped et al. and parasegment boundaries. AB - The pair-rule segmentation gene even skipped (eve) is required to activate engrailed stripes and to organize odd-numbered parasegments (PSs). The protein product Eve has been shown to be an active repressor of transcription, and recent models for Eve function suggest that activation of engrailed is indirect, but these models have not been fully tested. Here we identify the forkhead domain transcription factor Sloppy-paired as the key intermediate in the initial activation of engrailed by Eve in odd-numbered parasegments. We also analyze the roles of the transcription factors Runt and Odd-skipped in this process. Detailed analysis of engrailed and pair-rule gene expression in various mutant combinations shows how eve activates engrailed by repressing these engrailed repressors, and further indicates that mutual repression among pair-rule genes plays an important role in establishing parasegment boundaries. We present a new model of pair-rule gene function that explains the response of these boundaries to the relative levels of Eve and Fushi Tarazu. PMID- 15110724 TI - The semantics and acquisition of number words: integrating linguistic and developmental perspectives. AB - This article brings together two independent lines of research on numerally quantified expressions, e.g. two girls. One stems from work in linguistic theory and asks what truth conditional contributions such expressions make to the utterances in which they are used--in other words, what do numerals mean? The other comes from the study of language development and asks when and how children learn the meaning of such expressions. My goal is to show that when integrated, these two perspectives can both constrain and enrich each other in ways hitherto not considered. Specifically, work in linguistic theory suggests that in addition to their 'exactly n' interpretation, numerally quantified NPs such as two hoops can also receive an 'at least n' and an 'at most n' interpretation, e.g. you need to put two hoops on the pole to win (i.e. at least two hoops) and you can miss two shots and still win (i.e. at most two shots). I demonstrate here through the results of three sets of experiments that by the age of 5 children have implicit knowledge of the fact that expressions like two N can be interpreted as 'at least two N' and 'at most two N' while they do not yet know the meaning of corresponding expressions such as at least/most two N which convey these senses explicitly. I show that these results have important implications for theories of the semantics of numerals and that they raise new questions for developmental accounts of the number vocabulary. PMID- 15110725 TI - Baby hands that move to the rhythm of language: hearing babies acquiring sign languages babble silently on the hands. AB - The "ba, ba, ba" sound universal to babies' babbling around 7 months captures scientific attention because it provides insights into the mechanisms underlying language acquisition and vestiges of its evolutionary origins. Yet the prevailing mystery is what is the biological basis of babbling, with one hypothesis being that it is a non-linguistic motoric activity driven largely by the baby's emerging control over the mouth and jaw, and another being that it is a linguistic activity reflecting the babies' early sensitivity to specific phonetic syllabic patterns. Two groups of hearing babies were studied over time (ages 6, 10, and 12 months), equal in all developmental respects except for the modality of language input (mouth versus hand): three hearing babies acquiring spoken language (group 1: "speech-exposed") and a rare group of three hearing babies acquiring sign language only, not speech (group 2: "sign-exposed"). Despite this latter group's exposure to sign, the motoric hypothesis would predict similar hand activity to that seen in speech-exposed hearing babies because language acquisition in sign-exposed babies does not involve the mouth. Using innovative quantitative Optotrak 3-D motion-tracking technology, applied here for the first time to study infant language acquisition, we obtained physical measurements similar to a speech spectrogram, but for the hands. Here we discovered that the specific rhythmic frequencies of the hands of the sign-exposed hearing babies differed depending on whether they were producing linguistic activity, which they produced at a low frequency of approximately 1 Hz, versus non-linguistic activity, which they produced at a higher frequency of approximately 2.5 Hz - the identical class of hand activity that the speech-exposed hearing babies produced nearly exclusively. Surprisingly, without benefit of the mouth, hearing sign exposed babies alone babbled systematically on their hands. We conclude that babbling is fundamentally a linguistic activity and explain why the differentiation between linguistic and non-linguistic hand activity in a single manual modality (one distinct from the human mouth) could only have resulted if all babies are born with a sensitivity to specific rhythmic patterns at the heart of human language and the capacity to use them. PMID- 15110726 TI - A holistic account of the own-race effect in face recognition: evidence from a cross-cultural study. AB - A robust finding in the cross-cultural research is that people's memories for faces of their own race are superior to their memories for other-race faces. However, the mechanisms underlying the own-race effect have not been well defined. In this study, a holistic explanation was examined in which Caucasian and Asian participants were asked to recognize features of Caucasian and Asian faces presented in isolation and in the whole face. The main finding was that Caucasian participants recognized own-race faces more holistically than Asian faces whereas Asian participants demonstrated holistic recognition for both own race and other-race faces. The differences in holistic recognition between Caucasian and Asian participants mirrored differences in their relative experience with own-race and other-race faces. These results suggest that the own race effect may arise from the holistic recognition of faces from a highly familiar racial group. PMID- 15110727 TI - Age-of-acquisition effects in visual word recognition: evidence from expert vocabularies. AB - Three experiments assessed the contributions of age-of-acquisition (AoA) and frequency to visual word recognition. Three databases were created from electronic journals in chemistry, psychology and geology in order to identify technical words that are extremely frequent in each discipline but acquired late in life. In Experiment 1, psychologists and chemists showed an advantage in lexical decision for late-acquired/high-frequency words (e.g. a psychologist responding to cognition) over late-acquired/low-frequency words (e.g. a chemist responding to cognition), revealing a frequency effect when words are perfectly matched. However, contrary to theories that exclude AoA as a factor, performance was similar for the late-acquired/high-frequency and early-acquired/low-frequency words (e.g. dragon) even though their cumulative frequencies differed by more than an order of magnitude. This last finding was replicated with geologists using geology words matched with early-acquired words in terms of concreteness (Experiment 2). Most interestingly, Experiment 3 yielded the same pattern of results in naming while controlling for imageability, a finding that is particularly problematic for parallel distributed processing models of reading. PMID- 15110728 TI - Bodies capture attention when nothing is expected. AB - Functional neuroimaging research has shown that certain classes of visual stimulus selectively activate focal regions of visual cortex. Specifically, cortical areas that generally and selectively respond to faces (Kanwisher, N., McDermott, J., & Chun, M. M. (1997). The fusiform face area: a module in human extrastriate cortex specialized for face perception. Journal of Neuroscience, 17(11), 4302-4311; Puce, A., Allison, T., Asgari, M., Gore, J. C., & McCarthy, G. (1996). Differential sensitivity of human visual cortex to faces, letterstrings, and textures: a functional magnetic resonance imaging study. Journal of Neuroscience, 16(16), 5205-5215.) and to the human body (Downing, P. E., Jiang, Y., Shuman, M., & Kanwisher, N. (2001). A cortical area selective for visual processing of the human body. Science, 293(5539), 2470-2473.) have recently been described using fMRI. A parallel body of research has focused on the ability of faces to "capture" the focus of attention, compared to other kinds of objects (Lavie, N., Ro, T., & Russell, C. (2003). The role of perceptual load in processing distractor faces. Psychological Science, 14(5), 510-515; Ro, T., Russell, C., & Lavie, N. (2001). Changing faces: a detection advantage in the flicker paradigm. Psychological Science, 12(1), 94-99; Vuilleumier, P. (2000). Faces call for attention: evidence from patients with visual extinction. Neuropsychologia, 38(5), 693-700.). The present study uses Mack and Rock's "inattentional blindness" paradigm to investigate whether unexpected, task irrelevant human body stimuli capture awareness when attention is occupied by a primary task (Mack, A., & Rock, I. (1998). Inattentional blindness. London: MIT Press). Silhouettes and stick figures of human bodies, and silhouettes of hands, were compared to control stimuli including object silhouettes, object stick figures, and scrambled silhouettes of bodies, body parts, and objects. Participants were significantly better able to detect a human figure relative to the control stimuli. These results suggest that the human body, like the face, may be prioritized for attentional selection. More generally, they are consistent with the proposal that the visual system assigns attentional priority to types of stimuli that are also represented in strongly selective cortical regions. PMID- 15110729 TI - Amodal processing of visual speech as revealed by priming. AB - This study investigated the linguistic processing of visual speech (video of a talker's utterance without audio) by determining if such has the capacity to prime subsequently presented word and nonword targets. The priming procedure is well suited for the investigation of whether speech perception is amodal since visual speech primes can be used with targets presented in different modalities. To this end, a series of priming experiments were conducted using several tasks. It was found that visually spoken words (for which overt identification was poor) acted as reliable primes for repeated target words in the naming, written and auditory lexical decision tasks. These visual speech primes did not produce associative or reliable form priming. The lack of form priming suggests that the repetition priming effect was constrained by lexical level processes. That priming found in all tasks is consistent with the view that similar processes operate in both visual and auditory speech processing. PMID- 15110730 TI - Long-term implications of early onset in bipolar disorder: data from the first 1000 participants in the systematic treatment enhancement program for bipolar disorder (STEP-BD). AB - BACKGROUND: Early onset of mood symptoms in bipolar disorder has been associated with poor outcome in many studies; however, the factors that might contribute to poor outcome have not been adequately investigated. METHODS: The first consecutive 1000 adult bipolar patients enrolled in the National Institute of Mental Health's Systematic Treatment Enhancement Program for Bipolar Disorder were assessed at study entry to determine details of their age of onset of mood symptoms. Clinical course, comorbidity, and functional status and quality of life were compared for groups with very early (age < 13 years), early (age 13-18 years), and adult (age > 18 years) onset of mood symptoms. RESULTS: Of 983 subjects in whom age of onset could be determined, 272 (27.7%) experienced very early onset, and 370 (37.6%) experienced early onset. Earlier onset was associated with greater rates of comorbid anxiety disorders and substance abuse, more recurrences, shorter periods of euthymia, greater likelihood of suicide attempts and violence, and greater likelihood of being in a mood episode at study entry. CONCLUSIONS: Very early or early onset of bipolar disorder might herald a more severe disease course in terms of chronicity and comorbidity. Whether early intervention might modify this risk merits further investigation. PMID- 15110731 TI - Acute left prefrontal transcranial magnetic stimulation in depressed patients is associated with immediately increased activity in prefrontal cortical as well as subcortical regions. AB - BACKGROUND: Focal prefrontal cortex repetitive transcranial magnetic stimulation (rTMS) was originally investigated as a potential antidepressant under the assumption that in depressed patients, prefrontal cortex stimulation would produce changes in connected limbic regions involved in mood regulation. METHODS: Fourteen adult patients with depression were scanned in a 1.5-T scanner using interleaved rTMS (1 Hz) applied on the left prefrontal cortex over 7.35 min. Images were analyzed with Statistical Parametric Mapping 2b and principal component analysis. RESULTS: Over the left prefrontal cortex, 1-Hz TMS was associated with increased activity at the site of stimulation as well as in connected limbic regions: bilateral middle prefrontal cortex, right orbital frontal cortex, left hippocampus, mediodorsal nucleus of the thalamus, bilateral putamen, pulvinar, and insula (t = 3.85, p <.001). Significant deactivation was found in the right ventromedial frontal cortex. CONCLUSIONS: In depressed patients, 1-Hz TMS at 100% motor threshold over the left prefrontal cortex induces activation underneath the coil, activates frontal-subcortical neuronal circuits, and decreases activity in the right ventromedial cortex. Further work is needed to understand whether these immediate changes vary as a function of TMS use parameters (intensity, frequency, location) and whether they relate to neurobiologic effects and antidepressant mechanisms of TMS. PMID- 15110732 TI - Major depression is associated with lower omega-3 fatty acid levels in patients with recent acute coronary syndromes. AB - BACKGROUND: Polyunsaturated fatty acids (PUFAs) are intrinsic cell membrane components and closely involved in neurotransmission and receptor function. Lower omega-3 levels are associated with increased risk of coronary artery disease (CAD), increases in cardiac events in CAD patients, and depression. We sought to examine relationships between depression and serum levels of omega-3 and omega-6 PUFAs in patients recovering from acute coronary syndromes (ACS). METHODS: We carried out a case-control study of serum PUFA levels and current major depression in 54 age- and sex-matched pairs approximately 2 months following ACS. RESULTS: Depressed patients had significantly lower concentrations of total omega 3 and docosahexaenoic acid (DHA), and higher ratios of arachidonic acid (AA) to DHA, AA to eicosapentaenoic acid (EPA), and n-3 to n-6 than controls. There were no baseline differences in any potential risk or protective factors for depression. CONCLUSIONS: Results are consistent with previous reports in depressed patients without CAD, and with literature concerning omega-3 levels and risk of CAD events. Dietary, genetic, and hormonal factors may all play a role in both depression and CAD. Both prospective studies and randomized trials are needed to help clarify the interrelationships. PMID- 15110733 TI - Human amygdala responses during presentation of happy and neutral faces: correlations with state anxiety. AB - BACKGROUND: Previous functional imaging studies demonstrating amygdala response to happy facial expressions have all included the presentation of negatively valenced primary comparison expressions within the experimental context. This study assessed amygdala response to happy and neutral facial expressions in an experimental paradigm devoid of primary negatively valenced comparison expressions. METHODS: Sixteen human subjects (eight female) viewed 16-sec blocks of alternating happy and neutral faces interleaved with a baseline fixation condition during two functional magnetic resonance imaging scans. RESULTS: Within the ventral amygdala, a negative correlation between happy versus neutral signal changes and state anxiety was observed. The majority of the variability associated with this effect was explained by a positive relationship between state anxiety and signal change to neutral faces. CONCLUSIONS: Interpretation of amygdala responses to facial expressions of emotion will be influenced by considering the contribution of each constituent condition within a greater subtractive finding, as well as 1) their spatial location within the amygdaloid complex; and 2) the experimental context in which they were observed. Here, an observed relationship between state anxiety and ventral amygdala response to happy versus neutral faces was explained by response to neutral faces. PMID- 15110734 TI - [123I]AM281 single-photon emission computed tomography imaging of central cannabinoid CB1 receptors before and after Delta9-tetrahydrocannabinol therapy and whole-body scanning for assessment of radiation dose in tourette patients. AB - BACKGROUND: The cannabinoid CB1 receptor agonist Delta9-THC has been suggested for treatment of Tourette syndrome (TS). Based on animal studies, the CB1 antagonist [123I]AM281 (N-(Morpholin-4-yl)-1-(2,4-dichlorophenyl)-5-(4 [123I]iodophenyl)-4-methyl-1H-pyrazole-3-carboxamide) has been proposed for single photon emission computed tomography (SPECT) in humans. Our aims were to 1) evaluate specific binding of [123I]AM281 to CB1 receptors in TS patients and 2) assess radiation exposure associated with the use of AM281 labeled with 123I for SPECT and 124I for positron emission tomography. METHODS: We employed [123I]AM281 in six TS patients before and after Delta9-THC treatment. Dynamic SPECT, plasma measurements (including metabolite analysis with thin layer chromatography), and whole-body imaging were performed. Regions of interest derived from magnetic resonance images were used to extract from SPECT uptake in an area with high CB1 density (lentiform nuclei) and reference regions. Specific over nonspecific partition coefficients V3" were calculated. Whole-body images were carried out for dosimetric analysis. Data obtained with [123I]AM281 were used to predict doses from [124I]AM281. RESULTS: Mean V3" ranged from .19 to .31 and did not change significantly after Delta9-THC treatment. Nevertheless, in the only patient with a marked clinical response, V3" clearly declined. Thin layer chromatography revealed biexponential kinetics of tracer metabolism; about 60% remained nonmetabolized after 3 hours. Effective doses of .011 mSv/MBq for [123I]AM281 and .34 for [124I]AM281 were computed. CONCLUSIONS: This study suggests that specific binding of [123I]AM281 to CB1 receptors can be detected in patients using SPECT. Radiation exposure with [123I]AM281 is low; that with [124I]AM281 is higher but acceptable for single investigations. PMID- 15110735 TI - Cognitive-pharmacologic functional magnetic resonance imaging in tourette syndrome: a pilot study. AB - BACKGROUND: Dopamine agonists and antagonists can reduce abnormal movements and vocalizations (tics) in Tourette syndrome (TS); however, dopamine-responsive abnormal function in specific brain regions has not been directly demonstrated in TS. We sought to identify dopamine-modulated brain regions that function abnormally in TS by combining functional magnetic resonance imaging (fMRI), a working memory (WM) task, and infusion of the dopamine prodrug levodopa (while blocking dopamine production outside the brain). METHODS: We obtained complete fMRI data in 8 neuroleptic-naive adults with a chronic tic disorder and in 10 well-matched tic-free control subjects. RESULTS: Different task-sensitive brain regions responded differently to the WM task depending on levodopa status and diagnostic group (analysis of variance [ANOVA], p <.001). Four regions showed interactions with diagnosis (ANOVA, p <.001). In TS subjects, the task induced excessive brain activity in parietal cortex, medial frontal gyrus, and thalamus. Levodopa normalized the excess activity. In left parietal cortex, the degree of normalization was greater in patients with higher levodopa plasma concentrations (n = 6; Spearman's r = -.84, p =.04) and a greater degree of diagnostic confidence of TS (r = -.71, p =.05). CONCLUSIONS: These results are consistent with a dopamine-influenced functional abnormality of brain response in TS and suggest testable hypotheses about the mechanism by which dopamine antagonists and agonists alleviate tics. PMID- 15110736 TI - Development of left occipitotemporal systems for skilled reading in children after a phonologically- based intervention. AB - BACKGROUND: A range of neurobiological investigations shows a failure of left hemisphere posterior brain systems to function properly during reading in children and adults with reading disabilities. Such evidence of a disruption in the normal reading pathways provides a neurobiological target for reading interventions. In this study, we hypothesized that the provision of an evidence based, phonologically mediated reading intervention would improve reading fluency and the development of the fast-paced occipitotemporal systems serving skilled reading. METHODS: Functional magnetic resonance imaging was used to study the effects of a phonologically based reading intervention on brain organization and reading fluency in 77 children aged 6.1-9.4 years (49 with reading disability and 28 control subjects). Children comprised three experimental groups: experimental intervention (n = 37), community intervention (n = 12), and community control subjects (n = 28). RESULTS: Immediately after the year-long intervention, children taught with the experimental intervention had made significant gains in reading fluency and demonstrated increased activation in left hemisphere regions, including the inferior frontal gyrus and the middle temporal gyrus; 1 year after the experimental intervention had ended these children were activating bilateral inferior frontal gyri and left superior temporal and occipitotemporal regions. CONCLUSIONS: These data indicate that the nature of the remedial educational intervention is critical to successful outcomes in children with reading disabilities and that the use of an evidence-based phonologic reading intervention facilitates the development of those fast-paced neural systems that underlie skilled reading. PMID- 15110737 TI - Effects of sustained gamma-hydroxybutyrate treatments on spontaneous and evoked firing activity of locus coeruleus norepinephrine neurons. AB - BACKGROUND: Gamma-hydroxybutyrate is currently used to promote nighttime sleep in the treatment of narcolepsy; however, it is also a drug of abuse ("Liquid Ecstasy") associated with a withdrawal syndrome with anxiety features. Of interest, the activity of locus coeruleus neurons is a reflective index of these above mentioned behavioral states. METHODS: Using in vivo extracellular unitary recordings, sustained administration of gamma-hydroxybutyrate (40 mg/kg/day via minipump implanted subcutaneously) on the spontaneous and sensory-evoked burst firing of locus coeruleus norepinephrine neurons was assessed in rats. RESULTS: A 2-day and 10-day gamma-hydroxybutyrate administration decreased the spontaneous firing activity of locus coeruleus neurons by 52% and 54%, respectively, when compared with controls. A similar degree of attenuation on evoked burst firing of norepinephrine neurons also occurred in these rats (2-day gamma-hydroxybutyrate: 47% and 10-day gamma-hydroxybutyrate: 58%), when compared with controls. In contrast, rats treated with gamma-hydroxybutyrate for 10 days followed by removal of the minipump for 36 hours resulted in a 33% augmentation in spontaneous locus coeruleus activity as compared with controls. Furthermore, a robust 79% increase in burst firing in response to paw-pinch was exhibited in theses rats. CONCLUSIONS: Chronic gamma-hydroxybutyrate treatment inhibits the spontaneous and sensory-evoked burst firing of locus coeruleus norepinephrine neurons, whereas these indices are enhanced during drug withdrawal. The alteration in norepinephrine activity during chronic gamma-hydroxybutyrate administration may contribute to the ability of this agent to induce sleep and regulate narcoleptic episodes. Enhanced norepinephrine activity during withdrawal may be related to symptoms of anxiety on rapid termination of this drug in abusers. PMID- 15110738 TI - Basal and dexamethasone suppressed salivary cortisol concentrations in a community sample of patients with posttraumatic stress disorder. AB - BACKGROUND: Posttraumatic stress disorder (PTSD) has been associated with lower concentrations of cortisol and enhanced suppression of cortisol by dexamethasone, although discrepancies exist among reports. The objective of the study was to determine the pattern of cortisol responses in patients seeking treatment for PTSD resulting from a variety of traumatic experiences and to test whether cortisol responses are significantly related to childhood trauma, severity of symptoms, or length of time since trauma. METHODS: Salivary cortisol was measured at 8 AM, 4 PM, and 10 PM on 2 consecutive days before and after a 10 PM dose of .5 mg dexamethasone in 17 psychotropic medication and substance-free subjects with PTSD and 17 matched control subjects. RESULTS: Repeated-measures analysis of variance (ANOVA) of the baseline salivary cortisol concentrations demonstrated a significant effect for group with higher concentrations in the PTSD group but no significant differences in responses to dexamethasone. The presence of childhood abuse did not significantly affect salivary cortisol concentrations, and there was no correlation between predexamethasone cortisol and either the severity of PTSD symptoms or the time since the index trauma. CONCLUSIONS: Neither low basal concentrations nor enhanced suppression of cortisol are consistent markers of a PTSD diagnosis. PMID- 15110739 TI - A magnetic resonance imaging study of cortical thickness in animal phobia. AB - BACKGROUND: Despite the high prevalence of specific phobia (SP), its neural substrates remain undetermined. Although an initial series of functional neuroimaging studies have implicated paralimbic and sensory cortical regions in the pathophysiology of SP, to date contemporary morphometric neuroimaging methods have not been applied to test specific hypotheses regarding structural abnormalities. METHODS: Morphometric magnetic resonance imaging (MRI) methods were used to measure regional cortical thickness in 10 subjects with SP (animal type) and 20 healthy comparison (HC) subjects. RESULTS: Consistent with a priori hypotheses, between-group differences in cortical thickness were found within paralimbic and sensory cortical regions. Specifically, in comparison with the HC group, the SP group exhibited increased cortical thickness in bilateral insular, bilateral pregenual anterior cingulate, and bilateral posterior cingulate cortex as well as left visual cortical regions. CONCLUSIONS: Taken together, these structural findings parallel results from initial functional imaging studies that implicate paralimbic and sensory cortical regions in the mediating anatomy of SP symptoms. Further research will be necessary to replicate these findings and to determine their specificity as well as their pathophysiologic significance. PMID- 15110740 TI - Heart rate variability during sleep and the early development of posttraumatic stress disorder. AB - BACKGROUND: Noradrenergic function has been linked to posttraumatic stress disorder (PTSD) and might have a role in mediating sleep disturbances of the disorder. Our objective was to relate a peripheral manifestation of noradrenergic function, sympathetic nervous system activity as indexed by heart rate variability during sleep, to the development of PTSD in subjects with recent traumatic injuries. METHODS: Subjects who had recall of life-threatening experiences were recruited from one of two regional trauma centers. Select subjects received a polysomnographic recording within 1 month of the trauma. Digitized electrocardiogram recordings were extracted from early and late rapid eye-movement (REM) and preceding non-REM sleep periods. Autoregression was applied to R-R interval time series to calculate the ratios of low-frequency to high-frequency spectral densities (LF/HF ratios), which index sympathetic activation. Posttraumatic stress disorder status was determined at 2 months. RESULTS: There was a significant state x group interaction: LF/HF ratios were higher during the REM sleep of the nine subjects who were positive for PTSD symptoms, compared with the 10 subjects who were PTSD negative. CONCLUSIONS: Our findings are consistent with the possibility that increased noradrenergic activity during REM sleep contributes to the development of PTSD. PMID- 15110741 TI - Response to tryptophan depletion in major depression treated with either cognitive therapy or selective serotonin reuptake inhibitor antidepressants. AB - BACKGROUND: Mood sensitivity to rapid tryptophan depletion (RTD) has been demonstrated in patients treated with antidepressants that act preferentially on the serotonergic system. Depressed patients treated with bright-light therapy also show sensitivity to RTD, but those treated with electroconvulsive therapy or total sleep deprivation do not. Patients treated with an empirically supported psychotherapy have not been investigated for sensitivity to tryptophan depletion. This study compares the effects of RTD in patients treated with either selective serotonin re-uptake inhibitors (SSRIs) or cognitive therapy (CT). METHODS: Twenty patients treated with either SSRIs or CT underwent both rapid tryptophan depletion and sham-depletion using a blinded crossover design. Depressive symptoms were assessed using a modified Hamilton Depression Rating Scale (HDRS) and the Beck Depression Inventory (BDI). The differential change in depression scores across procedures between the groups was compared, and effect sizes were calculated. RESULTS: The differential worsening of mood for the SSRI group compared with the CT group was significant on the BDI. The effect size of the differential change was 1.6 for the BDI and.8 for the HDRS. Furthermore, the SSRI group experienced significant mood worsening during depletion compared with sham on both the HDRS and the BDI, whereas the CT group did not. CONCLUSIONS: The CT group was resistant to the effects of tryptophan depletion, but the SSRI group was not. PMID- 15110742 TI - Comment on "The identification of OCD-related subgroups based on comorbidity". PMID- 15110744 TI - Potent isothiocyanate inhibitors of carbonic anhydrase: synthesis and evaluation. AB - The reversible hydration of carbon dioxide by carbonic anhydrase (CA) regulates pH and carbon dioxide concentrations in diverse biological systems. Potent irreversible inhibition of CA would facilitate study of the dynamics of CA turnover as well as therapeutic effects due to long-term inhibition of the enzyme. We have synthesized isothiocyanate-containing sulfonamide inhibitors of CA from the corresponding aminosulfonamides. Significant increases in apparent binding of some of the isothiocyanate inhibitors over the amine analogues were consistent with covalent inhibition of the enzyme. PMID- 15110745 TI - Novel P143L polymorphism of the LCAT gene is associated with dyslipidemia in Chinese patients who have coronary atherosclerotic heart disease. AB - Coronary atherosclerotic heart disease (CAD) is a multifactorial disorder resulting from numerous gene-gene and gene-environment interactions. Lecithin:cholesterol acyltransferase (LCAT), a key enzyme in reverse cholesterol transport and the metabolism of high-density lipoprotein (HDL), is thought to be a candidate gene related to dyslipidemia and CAD. Variations in the LCAT gene were investigated in 190 CAD patients and 209 age- and gender-matched controls by denaturing high-performance liquid chromatography, and confirmed by sequencing and RFLP assay. In CAD patients, a novel single-nucleotide polymorphism (P143L) in exon 4 of the LCAT gene was discovered in nine males and two females (frequency of 5.79%), which was found in none of 209 controls. The genotype and allele distribution of P143L is significantly (P<0.04 ) higher in the low HDL-C subgroup than in the normal HDL-C subgroup in both male patients and all CAD patients. P143L was also found to be significantly (P<0.01) associated with the low HDL-C phenotype in both male patients and all CAD patients, with odds-ratios of 7.003 (95% CI 2.243-21.859) and 5.754 (95% CI 1.893-13.785), respectively. Thus, the P143L polymorphism may play a role in causing decreased HDL-C levels, leading to increased risk of dyslipidemia and CAD in Chinese. PMID- 15110746 TI - Groups on the side chain of T252 in Escherichia coli leucyl-tRNA synthetase are important for discrimination of amino acids and cell viability. AB - Leucyl-tRNA synthetase (LeuRS) catalyzes the leucylation of tRNA(Leu). To maintain the fidelity of protein biosynthesis, LeuRS also catalyzes the editing reaction. In the present work, highly conserved T252 in the T-rich region within CP1 domain of Escherichia coli LeuRS was mutated to G, D, or E. Steady-state kinetic of aminoacylation, and combined editing assays indicated that not only the size of the amino acid but also the absence of hydrogen bonds between T252 and adjacent molecules may affect the editing. It is further confirmed by in vivo experiments using the temperature-sensitive strain KL231 (DeltaleuS), which revealed the arrested growth of bacterial cells bearing mutants with highly impaired editing activity in the presence of leucine analog. PMID- 15110747 TI - Significance of the epsilon subunit in the thiol modulation of chloroplast ATP synthase. AB - To understand the regulatory function of the gamma and epsilon subunits of chloroplast ATP synthase in the membrane integrated complex, we constructed a chimeric FoF1 complex of thermophilic bacteria. When a part of the chloroplast F1 gamma subunit was introduced into the bacterial FoF1 complex, the inverted membrane vesicles with this chimeric FoF1 did not exhibit the redox sensitive ATP hydrolysis activity, which is a common property of the chloroplast ATP synthase. However, when the whole part or the C-terminal alpha-helices region of the epsilon subunit was substituted with the corresponding region from CF1-epsilon together with the mutation of gamma, the redox regulation property emerged. In contrast, ATP synthesis activity did not become redox sensitive even if both the regulatory region of CF1-gamma and the entire epsilon subunit from CF1 were introduced. These results provide important features for the regulation of FoF1 by these subunits: (1) the interaction between gamma and epsilon is important for the redox regulation of FoF1 complex by the gamma subunit, and (2) a certain structural matching between these regulatory subunits and the catalytic core of the enzyme must be required to confer the complete redox regulation mechanism to the bacterial FoF1. In addition, a structural requirement for the redox regulation of ATP hydrolysis activity might be different from that for the ATP synthesis activity. PMID- 15110748 TI - In vivo gene transfer into the adult honeybee brain by using electroporation. AB - The honeybee, Apis mellifera L., is a social insect and they show wide variety of exquisite social behaviors to maintain colony activity. To enable the elucidation of those social behaviors at a molecular level and gene function in the nervous system, we developed an in vivo method to perform gene transfer in the adult brain of living honeybee by electroporation. When green fluorescent protein expressing plasmid was transferred to the brain with this system, green fluorescence was observed near the anode location. The expression of transfected genes was confirmed at both transcriptional and translational levels by reverse transcription-polymerase chain reaction and immunoblot analyses. This system will facilitate the analysis of gene function and the regulatory mechanisms of gene networks in the nervous system and provide clues to clarify the relation between those genes and the complex behaviors of the honeybee. PMID- 15110749 TI - Macrophage phagocytosis of apoptotic neutrophils is compromised by matrix proteins modified by cigarette smoke and lipid peroxidation products. AB - Clearance of apoptotic cells by phagocytosis plays an important role in the resolution of an inflammatory response. Macrophages interacting with extracellular matrix (ECM) proteins upregulate their phagocytic capacity. Cigarette smoke contains highly reactive carbonyls that modify proteins which directly/indirectly affects cellular function. We observed, in vitro, that human macrophages interacting with carbonyl or cigarette smoke modified ECM proteins dramatically down regulated their ability to phagocytose apoptotic neutrophils. We also show that this interaction with carbonyl-adduct modified ECM proteins led to increased macrophage adhesion in vitro. We hypothesise that changes in the ECM environment as a result of cigarette smoking affect the ability of macrophages to remove apoptotic cells. Moreover, we postulate that this decreased phagocytic activity was as a result of sequestration of receptors involved in the uptake of apoptotic cells towards that of recognition of carbonyl adducts on the modified ECM proteins leading to increased macrophage adhesion. PMID- 15110750 TI - Characterization of a novel human gene containing ANK repeats and ARM domains. AB - We report here on characterization of a novel human gene on chromosome 2q32, containing several ankyrin repeats and ARM domains. The gene FLJ25415, including at least 31 exons and spanning about 90kb of a gene-rich genomic region, is present in the human genome as a single copy. It seems highly conserved along the mammalian phylogeny. By analysing FLJ25415 gene expression in different human tissues, we demonstrated a ubiquitously expression pattern with extensive alternative splicing. In silico prediction of promoter regions revealed the presence of a TATA box and some hypothetical transcription factor binding sites, such as TMF, C/EBPalpha, LE-1, and NF-ATp. PMID- 15110751 TI - Protein disulfide isomerase suppresses the transcriptional activity of NF-kappaB. AB - We report here that the transcriptional activity of NF-kappaB is negatively regulated by protein disulfide isomerase (PDI). Over-expression of PDI in RAW 264.7 cells strongly suppressed the LPS-induced production of inflammatory cytokines as well as NF-kappaB-dependent luciferase activity. This negative regulation of NF-kappaB was reversed by bacitracin, a PDI inhibitor. Interestingly, NF-kappaB/DNA complex formation and phosphorylation of NF-kappaB subunits was intact in PDI-expressing cells following stimulation with LPS. In addition, PDI and another redox regulator, thioredoxin (TRX), had opposite effects on NF-kappaB-dependent gene expression: activation of the NF-kappaB pathway by TRX was suppressed by expression of PDI in a dose-dependent manner. Finally, PDI expression was induced by the anti-inflammatory cytokine IL-10, and IL-10-mediated inhibition of LPS-induced IL-6 expression was reduced by bacitracin. These findings clearly demonstrate that PDI is a negative regulator of NF-kappaB, and may act downstream of IL-10 in this signaling pathway. PMID- 15110752 TI - High molecular mass kininogen inhibits metalloproteinases of Bothrops jararaca snake venom. AB - High molecular mass kininogen (HK) purified from Bothrops jararaca (Bj) plasma was tested on activities of the Bj venom in vivo and in vitro. Results showed that, when incubated with BjHK, the Bj venom presented inhibition on hemorrhagic, edema forming, myotoxic, and coagulant activities. It is well known that metalloproteinases are directly or indirectly involved in these activities. Similarly, human HK inhibits the hemorrhagic effect of the Bj venom as well as hemorrhagic and enzymatic effects of jararhagin, a hemorrhagic metalloproteinase isolated from Bj venom. Complex between HK and jararhagin was not detected by gel filtration. Nevertheless, the inhibitory effect of the hemorrhagic activity of the venom was only partial when HK was pre-incubated with 0.4mM ZnCl(2) or with 0.45mM CaCl(2). These data suggest that the inhibitory effect depends, at least partially, on the competition for ions between kininogen and metalloproteinases of the venom. PMID- 15110753 TI - Induction of immune activation by a novel immunomodulatory oligonucleotide without thymocyte apoptosis. AB - Bacterial DNA and synthetic oligodeoxynucleotides (ODN) containing unmethylated CpG motifs (CpG DNA) can potently stimulate innate immunity. While the actions of CpG DNA resemble those of LPS, these molecules stimulate distinct Toll-like receptors as well as cell types. In a previous study, we showed that a CpG ODN could induce cytokine production but, unlike LPS, did not induce thymocyte apoptosis. In this study, we have further investigated these differences using as a model a second-generation immunostimulatory oligonucleotide called HYB2048. Following administration to normal BALB/c mice, HYB2048-induced IL-12 but not IL 6 production. Under conditions in which LPS induced thymocyte apoptosis, HYB2048 did not cause significant cell death and, furthermore, did not block apoptosis induced by LPS. The levels of corticosterone induced by HYB2048 were also significantly lower than those induced by LPS. This pattern of activation could distinguish CpG DNA from LPS in its effects on the immune system. PMID- 15110754 TI - Effects of oligomycins on adenosine triphosphatase activity of mitochondria isolated from the yeasts Saccharomyces cerevisiae and Schwanniomyces castellii. AB - Functional mitochondria with respiratory control were isolated from the yeasts Saccharomyces cerevisiae and Schwanniomyces castellii. The presence of site I in Schw. castellii was indicated by higher ADP/O ratio than in S. cerevisiae where this site is absent. The ATPase Vmax was higher in S. cerevisiae than in Schw. castellii mitochondria. The latter was increased by the DR12 nuclear mutation. Nevertheless, the stimulation by heat and the inhibition profile of oligomycins on mitochondrial F1-F0 ATPase activities were similar in all three tested strains. In S. cerevisiae and Schw. castelli wild type or mutant mitochondria, the well-known inhibition of F1-F0 ATPase activity by low concentrations of oligomycins is abolished at high inhibitor concentrations near 60microg/ml suggesting uncoupling of F1 activity. At still higher oligomycin concentration the ATPase activity of both species and mutant is again strongly inhibited, suggesting an inhibitory effect on yeast F1 activity not detected so far. PMID- 15110755 TI - Cu+ distribution in metallothionein fragments. AB - The differential distribution of Cu+ between separate alpha and beta domains of metallothionein (the isolated peptide fragments) and the rate of transfer of Cu+ between the two domains using copper-thiolate specific emission spectroscopy are reported. Kinetic data show the rate of transfer of Cu+ from the Cu6alpha to the Cd3beta domain is 2 x 10(-1) s(-1) while the transfer from Cu6beta to the Cd4alpha domain is much slower at 8 x 10(-3) s(-1), indicating the greater binding affinity of Cu+ for the MT beta domain. We report that the emission intensity of Cu6beta is 0.45 the emission intensity of Cu6alpha-MT. Lambda(max) is shown to be a probe of the environment of the Cu+. A series of copper containing domain intermediates to the formation of the filled Cu6S9-beta and Cu6S11-alpha-clusters are identified. A mechanism is proposed for the formation of Cu12(betaalpha)-MT that involves metal exchange reactions of Cu+ ions from the alpha to the beta domain with initial formation of a Cu4beta-cluster. PMID- 15110756 TI - Involvement of colicin in the limited protection of the colicin producing cells against bacteriophage. AB - The restriction/modification system is considered to be the most common machinery of microorganisms for protection against bacteriophage infection. However, we found that mitomycin C induced Escherichia coli containing ColE7-K317 can confer limited protection against bacteriophage M13K07 and lambda infection. Our study showed that degree of protection is correlated with the expression level of the ColE7 operon, indicating that colicin E7 alone or the colicin E7-immunity protein complex is directly involved in this protection mechanism. It was also noted that the degree of protection is greater against the single-strand DNA bacteriophage M13K07 than the double-strand bacteriophage(lambda). Coincidently, the K(A) value of ColE7-Im either interacting with single-strand DNA (2.94x10(5)M(-1)) or double strand DNA (1.75x10(5)M(-1)) reveals that the binding affinity of ColE7-Im with ssDNA is 1.68-fold stronger than that of the protein complex interacting with dsDNA. Interaction between colicin and the DNA may play a central role in this limited protection of the colicin-producing cell against bacteriophages. Based on these observations, we suggest that the colicin exporting pathway may interact to some extent with the bacteriophage infection pathway leading to a limited selective advantage for and limited protection of colicin-producing cells against different bacteriophages. PMID- 15110757 TI - Heme oxygenase-2 gene deletion increases astrocyte vulnerability to hemin. AB - In a prior study, we observed that heme oxygenase-2 gene deletion protected murine cortical neurons from heme-mediated injury. In the course of these studies, constitutive HO-2 expression was observed in astrocyte cultures. The present study tested the hypothesis that astrocytes lacking the HO-2 gene would be less vulnerable to heme. Contrary to this hypothesis, gene deletion resulted in a 50-75% increase in cell death after 6h exposure to 30 or 60microM hemin, as measured by LDH release. A similar effect was observed when cell viability was assessed with the MTT assay. HO-2 gene deletion did not alter cellular expression of HO-1. The increased sensitivity of knockout astrocytes to hemin was reversed by increasing HO-1 expression by adenoviral gene transfer. These results suggest that heme oxygenase protects astrocytes from heme-mediated oxidative injury and highlight the disparate effect of HO in neurons and astrocytes. PMID- 15110758 TI - FAST is a BCL-X(L)-associated mitochondrial protein. AB - The TIA-1-interacting protein Fas-activated serine/threonine phosphoprotein (FAST) is a component of a signaling cascade that is initiated by ligation of the Fas receptor. Immunofluorescence microscopy using affinity-purified antibodies raised against recombinant FAST reveals that the endogenous protein associates with mitochondria. Subcellular fractionation confirms that FAST is a component of mitochondria. FAST is tethered to mitochondria by a lysine/arginine-rich domain at its carboxyl terminus that is structurally similar to the mitochondrial tethering motifs of monoamine oxidase B and cytochrome b5. At the mitochondrial membrane, FAST interacts with BCL-X(L). The BCL-X(L) binding domain maps to a BCL 2-homology-3 (BH3)-related domain that is distinct from the mitochondrial tethering domain (MTD). Although interactions between FAST and BCL-X(L) require both the BH3-related domain and the MTD, the requirement for mitochondrial tethering can be conferred by a heterologous MTD. Our results suggest that FAST BCL-X(L) interactions are likely to regulate mitochondrial metabolism during Fas induced apoptosis. PMID- 15110759 TI - 17Beta-estradiol induces nuclear translocation of CrkL at the window of embryo implantation. AB - Crk family adaptors are widely expressed and mediate the timely formation of signal transduction protein complexes upon a variety of extracellular stimuli, including various growth and differentiation factors. The window of implantation is the favorable time period when the uterus develops a receptive approach to the invading embryo. Various signaling cascades are likely to become active at the window of implantation both in the uterus and the embryo. This helps create maternal embryo dialogue leading to successful embryo implantation. In this study we report for the first time the presence and nuclear translocation of the adaptor molecule CrkL both in the uterine and embryonic partners at the window of implantation. We also report that estrogen, which initiates and guides crucial changes in the uterus and the embryo at the window of receptivity, causes a massive surge in the expression and subsequent nuclear translocation of CrkL. We have also identified the existence of one LXXLL motif in the CrkL amino acid sequence and a single LXD is sufficient for activation by the estrogen receptor. This is suggestive that CrkL can bind to estrogen receptors and act as a coactivator. PMID- 15110761 TI - Isolation and enrichment of skeletal muscle progenitor cells from mouse bone marrow. AB - There is great interest in the therapeutic potential of non-hematopoietic stem cells obtained from bone marrow called mesenchymal stem cells (MSCs). Rare myogenic progenitor cells in MSC cultures have been shown to convert into skeletal muscle cells in vitro and also in vivo after transplantation of bone marrow into mice. To be clinically useful, however, isolation and expansion of myogenic progenitor cells is important to improve the efficacy of cell transplantation in generating normal skeletal muscle cells. We introduced into MSCs obtained from mouse bone marrow, a plasmid vector in which an antibiotic (Zeocin) resistance gene is driven by MyoD and Myf5 enhancer elements, which are selectively active in skeletal muscle progenitor cells. Myogenic precursor cells were then isolated by antibiotic selection, expanded in culture, and shown to differentiate appropriately into multinucleate myotubes in vitro. Our results show that using a genetic selection strategy, an enriched population of myogenic progenitor cells, which will be useful for cell transplantation therapies, can be isolated from MSCs. PMID- 15110760 TI - BDNF is induced by wild-type alpha-synuclein but not by the two mutants, A30P or A53T, in glioma cell line. AB - Parkinson's disease (PD) is one of the most prevalent neurodegenerative diseases but its etiology is unclear. Alpha-synuclein (alpha-SN) is a major component of Lewy bodies and Lewy neurites, and its missense mutations, A30P and A53T, cause familial PD. In PD, alpha-SN-positive glial inclusions are distributed mainly in the dorso-medial region of the substantia nigra, which contains most of the surviving dopaminergic neurons, suggesting that alpha-SN expression might have a neuroprotective function in glial cells. To investigate this hypothesis, we established alpha-SN transfected C6 glioma cell line clones and evaluated the expression of neurotrophins using semi-quantitative reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay. Brain-derived neurotrophic factor (BDNF) was induced by overexpression of wild-type alpha-SN but not by that of A30P and A53T. These data suggest that the pathogenic alpha-SN mutations, A30P or A53T, are linked to the loss of BDNF production in glial cells. PMID- 15110762 TI - Passage of murine scrapie prion protein across the mouse vascular blood-brain barrier. AB - Prions are the infectious agents associated with transmissible spongiform encephalopathies and are composed mainly of a misfolded form of the endogenous prion protein. Prion protein must enter the brain to produce disease. Previous work has emphasized various mechanisms which partially bypass the blood-brain barrier (BBB). Here, we used the brain perfusion method to directly assess the ability of mouse scrapie protein (PrP(SC)) to cross the mouse BBB independent of the influences of neural pathways or circulating immune cells. We found that PrP(SC) oligomers rapidly crossed the BBB without disrupting it with a unidirectional influx rate of about 4.4microl/g-min. HPLC and capillary depletion confirmed that PrP(SC) crossed the entire width of the capillary wall to enter brain parenchyma. PrP(SC) also entered the cerebrospinal fluid (CSF) compartment. These results show that a prion protein can cross the intact BBB to enter both the parenchymal and CSF compartments of the brain. PMID- 15110763 TI - The derivation of a potential transition state for the reduction reaction catalysed by 17beta-hydroxysteroid dehydrogenase--an approximate representation of its active site for use in drug design and discovery. AB - In an effort to aid the drug discovery process, the approximate representation of the active site of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) has been derived from the consideration of the proposed mechanism for the reduction reaction. Using the transition-state (TS), the mode of action of a number of inhibitors has been rationalised. The results of the study suggest that the area of the active site corresponding to the C(17) area of the steroidal backbone is relatively constrained, as such, groups which possess large groups that are flexible in nature may possess a decreased inhibitory activity. The model is therefore a good start point for the development of novel inhibitors of 17beta HSD and is a rapid technique for drug design and development. PMID- 15110764 TI - H2O2 regulates recombinant Ca2+ channel alpha1C subunits but does not mediate their sensitivity to acute hypoxia. AB - Acute hypoxic inhibition of the pore-forming alpha(1C) subunit of the L-type Ca(2+) channel mediates hypoxic arterial vasodilatation, a physiological response which matches tissue O(2) demand and supply in the systemic vasculature. In numerous O(2)-sensing cell types, reactive O(2) species (ROS) have been proposed as mediators linking lowered O(2) levels with the appropriate cellular response. In this study, we examined the roles of H(2)O(2) and NADPH oxidase as mediators of hypoxic inhibition of recombinant alpha(1C) subunits. Human cardiac L-type Ca(2+) channel alpha(1C) subunits were stably expressed in HEK 293 cells. Ca(2+) currents were recorded using the whole-cell configuration of the patch-clamp technique. Bath application of 100microM H(2)O(2) significantly enhanced depolarisation-evoked Ca(2+) currents in a voltage-dependent manner, while dialysis with 1000Uml(-1) catalase reduced these currents. In the presence of catalase, hypoxic inhibition of Ca(2+) currents was not significantly different compared to non-dialysed controls. The NADPH oxidase inhibitors diphenylene iodonium (10microM) and phenylarsine oxide (5microM) were without effect on either basal Ca(2+) currents or responses to hypoxia. Thus, endogenous production of H(2)O(2) regulates the alpha(1C) subunit. However, neither suppression of H(2)O(2) levels nor inhibition of NADPH oxidase is involved in O(2)-dependent regulation of the Ca(2+) channel. PMID- 15110765 TI - Prediction of the disulfide-bonding state of cysteines in proteins based on dipeptide composition. AB - In this paper, a novel approach has been introduced to predict the disulfide bonding state of cysteines in proteins by means of a linear discriminator based on their dipeptide composition. The prediction is performed with a newly enlarged dataset with 8114 cysteine-containing segments extracted from 1856 non-homologous proteins of well-resolved three-dimensional structures. The oxidation of cysteines exhibits obvious cooperativity: almost all cysteines in disulfide-bond containing proteins are in the oxidized form. This cooperativity can be well described by protein's dipeptide composition, based on which the prediction accuracy of the oxidation form of cysteines scores as high as 89.1% and 85.2%, when measured on cysteine and protein basis using the rigorous jack-knife procedure, respectively. The result demonstrates the applicability of this new relatively simple method and provides superior prediction performance compared with existing methods for the prediction of the oxidation states of cysteines in proteins. PMID- 15110766 TI - Spatio-temporal organization of Vam6P and SNAP on mouse spermatozoa and their involvement in sperm-zona pellucida interactions. AB - Acrosomal assembly during spermatogenesis and acrosome reaction during sperm oocyte interaction are unique events of vesicle synthesis, transport, and fusion leading to fertilization. SNARE complex formation is essential for membrane fusion, and vesicle-associated (v-) SNARE intertwines with target membrane (t-) SNARE to form a coiled coil that bridges two membranes and facilitates fusion. We detected messages of Vam6P and SNAP in mammalian testis and epididymis. Vam6P and SNAP were detected in a temporally organized fashion on the spermatozoa from testis and epididymis, which showed accumulation on the principal acrosomal domains during capacitation. Vam6P and SNAP were shed off from the principal acrosomal domain after acrosome reaction, but the equatorial and the post acrosomal domains retained these proteins. Antibodies to VAMP and SNAP inhibited sperm-zona pellucida interaction, suggesting their possible involvement in sperm membrane vesiculation. PMID- 15110767 TI - The gut-brain peptide neuromedin U is involved in the mammalian circadian oscillator system. AB - Immunohistochemical analysis revealed the presence of a gut-brain peptide, neuromedin U (NMU), in the suprachiasmatic nucleus (SCN), which is the site of the master circadian oscillator. The expression of NMU mRNA exhibited a circadian rhythm, with the peak expression in the SCN occurring at CT4-8h. The two NMU binding receptors (NMU-R1 and NMU-R2) were also expressed in the SCN, but their phase angles were different. Intracerebroventricular injection (ICV) of NMU induced the expression of Fos protein in the SCN cells and caused a phase dependent phase shift of the circadian locomotor activity rhythm. The magnitude of the phase shift was dose dependent. This NMU-induced phase shift was of the nonphotic type. Quantitative reverse transcriptase-polymerase chain reaction (RT PCR) analysis revealed increases in the expression in the SCN of immediate early genes, such as c-fos, NGFI-A, NGFI-B, and JunB. Furthermore, ICV injection of NMU increased the expression of Per1, but not Per2, in the SCN. These results indicate that NMU may play some important role in the circadian oscillator by exerting an autocrine or paracrine action in the SCN. PMID- 15110768 TI - Effects of basic fibroblast growth factor on osteoclasts and osteoclast-like cells. AB - Mouse marrow, which contains osteoblast and osteoclast precursors, was grown in the presence of calcitriol and/or basic fibroblast growth factor (FGF-2). RAW 264.7 cells were differentiated into osteoclast-like cells in the presence of receptor activator of NF-kappaB-Ligand (RANK-L) and/or FGF-2. FGF-2 alone supported osteoclastogenesis in mouse marrow cultures, but not by RAW 264.7 cells alone. Although FGF-2 supported low levels of osteoclastogenesis in mouse marrow cultures, it strongly inhibited the high levels of osteoclastogenesis triggered by calcitriol. Adding excess recombinant-RANK-L to the cultures did not relieve this inhibition. After mouse marrow osteoclasts were differentiated, FGF-2 dose dependently inhibited bone resorptive activity. FGF-2 increased the tendency of RAW 264.7 osteoclast-like cells to fuse into very large giant cells and induced reorganizations of the actin cytoskeleton in mature, RANK-L-induced RAW 264.7 osteoclast-like cells. These results suggest that FGF-2 has both direct and indirect effects on osteoclast formation and bone resorption. PMID- 15110769 TI - Targets for TNF-alpha-induced lipolysis in human adipocytes. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha)-induced lipolysis may be important for insulin resistance in both obesity and cachexia. In rodent cells TNF-alpha enhances lipolysis through down-regulation of the expression of the membrane proteins Galpha(i) and the lipid droplet-associated protein perilipin (PLIN). In human (but not murine) adipocytes TNF-alpha stimulates lipolysis through the mitogen activated protein kinases (MAPKs) p44/42 and JNK although it is unclear whether this is mediated via PLIN and/or Galpha(i). METHODS: Galpha(i) and PLIN as down-stream effectors of MAPKs were assessed in human adipocytes stimulated with TNF-alpha in the absence or presence of specific MAPK inhibitors. RESULTS: A 48-h incubation with TNF-alpha resulted in a pronounced increase in lipolysis, which was paralleled by a decrease in the mRNA and protein expression of PLIN. Both these effects were inhibited in a concentration-dependent manner in the presence of MAPK inhibitors specific for p44/42 (PD98059) and JNK (SP600125). However, TNF-alpha did not affect Galpha(i) mRNA or protein expression. Furthermore, experiments with pertussis toxin demonstrated that inhibition of Galpha(i) signaling did not affect TNF-alpha-mediated lipolysis. CONCLUSIONS: Our results suggest that TNF-alpha-mediated lipolysis is dependent on down-regulation of PLIN expression via p44/42 and JNK. This could be an important mechanism for the development of insulin resistance in both obesity and cachexia. However, in contrast to findings in rodent cells, Galpha(i) does not appear to be essential for TNF-alpha-induced lipolysis in human adipocytes. PMID- 15110770 TI - Comparative gene expression in the symbiotic and aposymbiotic Aiptasia pulchella by expressed sequence tag analysis. AB - Intracellular symbiotic relationships are prevalent between cnidarians, such as corals and sea anemones, and the photosynthetic dinoflagellate symbionts. However, there is little understanding about how the genes express when the symbiotic relationship is set up. To characterize genes involved in this association, the endosymbiosis between sea anemone, Aiptasia pulchella, and dinoflagellate zooxanthellae, Symbiodinium spp., was employed as a model. Two complementary DNA (cDNA) libraries were constructed from RNA isolated from symbiotic and aposymbiotic A. pulchella. Using single-pass sequencing of cDNA clones, a total of 870 expressed sequence tags (ESTs) clones were generated from the two libraries: 474 from symbiotic animal and 396 from aposymbiotic animal. The initial ESTs consisted of 143 clusters and 231 singletons. A BLASTX search revealed that 147 unique genes had similarities with protein sequences available from databases; 120 of these clones were categorized according to their putative function. However, many ESTs could not assign functionally. The putative roles of some of the identified genes relative to endosymbiosis were discussed. This is the first report of the use of EST analysis to examine the gene expression in symbiotic and aposymbiotic states of the cnidarians. The systematic analysis of EST from this study provides a useful database for future investigations of the molecular mechanisms involved in algal-cnidarian symbiosis. PMID- 15110771 TI - A genetic variant of ACE increases cell survival: a new paradigm for biology and disease. AB - The human angiotensin converting enzyme (ACE) polymorphism is caused by an Alu element insertion resulting in three genotypes (Alu+/+, Alu+/-, Alu-/-, or ACE II, ACE-ID, and ACE-DD, respectively), with ACE-II displaying lower ACE activity. The polymorphism is associated with athletic performance, aging, and disease. Population studies, however, were confounding because variants of the polymorphism appeared to fortuitously correlate with health and various pathological states. To clarify the functional role of the polymorphism, we studied its direct effect on cell survival. ACE-II (Alu+/+) human endothelial cells (EC) had lower angiotensin-II levels and 20-fold increased viability after slow starvation as compared to ACE-DD cells (Alu-/-). By RT-PCR, only ACE-II cells expressed the pluripotent/stem cell-maintenance factors nanog, numb, and klotho. ACE inhibition by captopril in ACE-DD cells mimicked the ACE-II genotype. These results provide the first evidence of a functional role for a naturally occurring polymorphism, having broad implications for human biology, longevity, and disease. PMID- 15110772 TI - Exogenous nitric oxide activates the endothelial glucocorticoid receptor. AB - This study investigated the effect of exogenous nitric oxide (NO) on endothelial glucocorticoid receptor (GR) function. The NO donor diethylenetriamine NONOate (DETA, 50-500microM) caused concentration dependent nuclear localization of transfected chimeric green fluorescent protein GFP-GR and elevated expression of secreted alkaline phosphatase (SEAP) from a glucocorticoid response element (GRE) promoter construct in bovine aortic endothelial cells. Other weaker NO donors (S nitroso-N-acetylpenicillamine and spermine NONOate) failed to induce GFP-GR nuclear localization, but all the NO donors activated GRE-SEAP expression, a response unaffected by the antioxidant N-acetyl-L-cysteine. Overall, exogenous NO from high concentration donors can directly activate GR, suggesting a potential feedback mechanism for NO to regulate endothelial inducible nitric oxide synthase (iNOS) expression. PMID- 15110773 TI - In vivo interaction between the human dehydrodolichyl diphosphate synthase and the Niemann-Pick C2 protein revealed by a yeast two-hybrid system. AB - Dehydrodolichyl diphosphate (DedolPP) synthase catalyzes the sequential condensation of isopentenyl diphosphate with farnesyl diphosphate to synthesize DedolPP, a biosynthetic precursor for dolichol which plays an important role as a sugar-carrier lipid in the biosynthesis of glycoprotein in eukaryotic cells. During certain pathological processes like Alzheimer's disease or some neurological disorders, dolichol has been shown to accumulate in human brain. In order to understand the regulatory mechanism of dolichol in eukaryotes, we performed a yeast two-hybrid screen using full length human DedolPP synthase gene [Endo et al. BBA 1625 (2003) 291] as a bait to find some proteins specifically interacting with the enzyme. We identified Niemann-Pick Type C2 protein (NPC2) to show a specific interaction with human DedolPP synthase. This interaction was further confirmed by in vitro co-immunoprecipitation experiment, indicating the possible physiological interaction between NPC2 and DedolPP synthase proteins in human. PMID- 15110774 TI - Tyrosine phosphorylated Disabled 1 recruits Crk family adapter proteins. AB - Disabled 1 (Dab1) functions as a critical adapter protein in the Reelin signaling pathway to direct proper positioning of neurons during brain development. Reelin stimulates phosphorylation of Dab1 on tyrosines 198 and 220, and phosphorylated Dab1 is likely to interact with downstream signaling proteins that contain Src homology 2 (SH2) domains. To search for such proteins, we used a Sepharose conjugated peptide containing phosphotyrosine 220 (PTyr-220) of Dab1, as an affinity matrix to capture binding proteins from mouse brain extracts. Mass spectrometric analysis of bound proteins revealed that Crk family adapter proteins selectively associated with this phosphorylation site. We further show that Crk-I and Crk-II, but not CrkL, stimulate phosphorylation of Dab1 on tyrosine 220 in a Src-dependent manner. Our results suggest that Crk family adapter proteins may play an important role in the Reelin signaling pathway during brain development. PMID- 15110775 TI - Sequential gene promoter interactions by C/EBPbeta, C/EBPalpha, and PPARgamma during adipogenesis. AB - Treatment of 3T3-L1 preadipocytes with differentiation inducers triggers a cascade in which C/EBPbeta is rapidly expressed, followed by C/EBPalpha and PPARgamma. C/EBPalpha and PPARgamma then activate the expression of adipocyte genes that produce the differentiated phenotype. Circumstantial evidence indicates that C/EBPbeta activates transcription of the C/EBPalpha and PPARgamma genes, both of which possess C/EBP regulatory elements in their proximal promoters. Although C/EBPbeta is expressed immediately upon induction of differentiation, acquisition of DNA binding activity is delayed for approximately 14h. Chromatin immunoprecipitation (ChIP) analysis conducted 24h after induction revealed that C/EBPbeta binds to C/EBP regulatory elements in the proximal promoters of the C/EBPalpha and PPARgamma genes. ChIP analysis showed that after an additional delay C/EBPalpha binds to its own promoter and to the promoters of the PPARgamma and 422/aP2 genes. These findings support the view that once expressed, C/EBPalpha is responsible for maintaining the expression of PPARgamma, and C/EBPalpha, as well as adipocyte proteins (e.g., 422/aP2) in the terminally differentiated state. Together these findings provide compelling evidence that C/EBPbeta, C/EBPalpha, and PPARgamma participate in a cascade during adipogenesis. PMID- 15110776 TI - Ouabain treatment is associated with upregulation of phosphatase inhibitor-1 and Na+/Ca(2+)-exchanger and beta-adrenergic sensitization in rat hearts. AB - Cardiac glycosides are widely used in the treatment of congestive heart failure. While the mechanism of the positive inotropic effect after acute application of cardiac glycosides is explained by blockade of the Na+/K+-pump, little is known about consequences of a prolonged therapy. Here male Wistar rats were treated for 4 days with continuous infusions of ouabain (6.5 mg/kg/day) or 0.9% NaCl (control) via osmotic minipumps. Electrically driven (1 Hz, 35 degrees C) papillary muscles from ouabain-treated rats exhibited shorter relaxation time ( 15%) and a twofold increase in the sensitivity for the positive inotropic effect of isoprenaline. The density and affinity of beta1- and beta2-adrenoceptors as well as mRNA and protein levels of stimulatory (G(s)alpha) and inhibitory (G(i)alpha-2, G(i)alpha-3) G-proteins were unaffected by ouabain. Similarly, SR Ca2+-ATPase 2A, phospholamban, ryanodine-receptor expression as well as the oxalate-stimulated 45Ca-uptake of membrane vesicles remained unchanged. However, mRNA abundance of the protein phosphatase inhibitor-1 (I-1) and the Na+/Ca2+ exchanger (NCX) were increased by 52% and 26%, respectively. I-1 plays an amplifier role in cardiac signaling. Downregulation of I-1 in human heart failure is associated with desensitization of the beta-adrenergic signaling pathway. The present data suggest that the ouabain-induced increase in I-1 expression might be at least partly responsible for the increased isoprenaline sensitivity and increased expression of NCX for the accelerated relaxation after chronic ouabain in this model. PMID- 15110777 TI - Characterization of recombinant soluble carcinoembryonic antigen cell adhesion molecule 1. AB - Carcinoembryonic antigen cell adhesion molecule 1 (CEACAM1) is a type 1 transmembrane, homotypic cell adhesion protein expressed on epithelial and hematopoietic cells. CEACAM1 has four major isoforms with three or four immunoglobulin (Ig)-like ectodomains and either long or short cytoplasmic domains. In a 3D model of breast epithelial cell morphogenesis, CEACAM1 plays an essential role in lumen formation [J. Cell Sci. 112 (1999) 4193]. Two soluble ectodomain isoforms of CEACAM1 expressed in myeloma cells were immunologically active and highly glycosylated. The molecular weights of the 3-ecto- and 4 ectodomain isoforms were 90 and 110kDa, respectively, and monomers by sedimentation equilibrium centrifugation. Both isoforms were prolate ellipsoids with axial ratios of 6 for the 3-ecto- and 8 for 4-ectodomain isoforms, respectively, by size exclusion chromatography and analytical ultracentrifugation. Both isoforms caused a significant reduction in lumen formation when tested in the 3D model culture system. PMID- 15110778 TI - Gene expression changes in rat white adipose tissue after a high-fat diet determined by differential display. AB - The differences in gene expression pattern of visceral white adipose tissue between control and high-fat-fed rats were compared using the mRNA differential display methodology. The results, confirmed by Northern blot, showed eight genes upregulated: adiponectin, fibrillin-1, transferrin, Y-box binding protein-1, IgE receptor beta chain (FcRIbeta), alpha-1 haemoglobin, and ribosomal proteins S10 and L7 and four genes downregulated: caveolin-2, lactate dehydrogenase-A, mitochondrial 16S rRNA, and mitochondrial cytochrome oxidase subunit I/serine tRNA. Two of these genes have been already related to obesity (adiponectin and caveolin-2) while the others are known to participate in metabolic, signalling or transcription regulation pathways that can be relevant in energy (lipid and/or carbohydrate) metabolism. PMID- 15110779 TI - Shared pathways of osteoblast mitogenesis induced by amylin, adrenomedullin, and IGF-1. AB - Amylin and adrenomedullin, members of the calcitonin peptide family, are anabolic to bone. Here, we report overlapping molecular mechanisms by which amylin, adrenomedullin, and IGF-1 induce osteoblast proliferation. Co-treatment of osteoblastic cells with amylin or adrenomedullin and IGF-1 failed to induce an additive mitogenic effect. In osteoblastic cells, neutralization of the IGF-1 receptor blocked the proliferative effects of amylin and adrenomedullin, while neutralization of IGF-1 did not. Neither amylin- nor adrenomedullin-induced mitogenic signaling or cell proliferation in IGF-1 receptor-null fibroblasts. In addition, amylin and adrenomedullin receptor blockers inhibited the proliferative effects of IGF-1 in osteoblastic cells. These findings demonstrate overlap in the molecular mechanisms by which amylin, adrenomedullin, and IGF-1 induce mitogenesis in osteoblasts, and an important role for the IGF-1 receptor in the mitogenic actions of amylin and adrenomedullin. Our findings are potentially important in refining these peptides for the therapy of osteoporosis. PMID- 15110781 TI - Protein profile of aging and its retardation by caloric restriction in neural retina. AB - Aging is a slow, gradual deterioration process of an organism. The only experimental intervention, which can reliably retard aging and age-related degenerative diseases, is dietary caloric restriction (CR). To gain insight into the mechanism of CR intervention, we have investigated the protein profile of aging and its retardation by CR in the neural retina of Brown Norway (BN) rats using the comprehensive proteomic approach. We found that the intensities of 18 proteins decreased significantly with age. CR intervention can completely prevent seven of them, and partially protect eight of them, from such age-related declines. The major protein targets protected by CR intervention appear to be glycolytic enzymes and molecular chaperones. These data are the first to suggest that CR may retard the age-related degeneration of retina by maintaining sufficient glucose metabolism, by ensuring proper protein folding, and/or by preventing protein denaturation in the neural retina. PMID- 15110780 TI - Salivary phospholipid secretion in response to beta-adrenergic stimulation is mediated by Src kinase-dependent epidermal growth factor receptor transactivation. AB - Communication between receptor tyrosine kinase and G protein-coupled receptor (GPCR)-mediated signaling is recognized as a common integrator linking diverse aspects of intracellular signaling systems. Here, we report that G protein coupled beta-adrenergic receptor activation leading to stimulation of salivary phospholipid release occurs with the involvement of epidermal growth factor receptor (EGFR). Using sublingual gland acinar cells, we show that prosecretory effect of isoproterenol on phospholipid release was subjected to suppression by EGFR kinase inhibitor, PD153035, and wortmannin, an inhibitor of PI3K, but not by PD98059, an inhibitor of extracellular signal regulated kinase (ERK). Furthermore, wortmannin, but not the ERK inhibitor, caused the reduction in the acinar cell secretory responses to beta-adrenergic agonist-generated cAMP as well as adenyl cyclase activator, forskolin. The acinar cell phospholipid secretory responses to isoproterenol, moreover, were inhibited by PP2, a selective inhibitor of tyrosine kinase Src responsible for ligand-independent EGFR phosphorylation. Taken together, our data are the first to demonstrate the requirement for Src kinase-dependent EGFR transactivation in regulation of salivary phospholipid secretion in response to beta-adrenergic GPCR activation. PMID- 15110782 TI - Glucocorticoid enhances the expression of dickkopf-1 in human osteoblasts: novel mechanism of glucocorticoid-induced osteoporosis. AB - To clarify the underlying mechanism of glucocorticoid-induced osteoporosis, we investigated the effect of glucocorticoid on the expression of dickkopf-1 (Dkk 1), an antagonist of Wnt signaling, in primary cultured human osteoblasts. Dexamethasone markedly induced the expression of mRNA for Dkk-1 in a dose- and time-dependent manner. The expression of Kremen1, a receptor for Dkk, did not change by the treatment with dexamethasone, while that of low-density lipoprotein receptor-related protein 5 (LRP5), a Wnt coreceptor, slightly decreased by the treatment with dexamethasone. Dexamethasone increased the transcriptional activity of the Dkk-1 gene promoter in human osteoblasts. Serial deletion and mutation analyses of the Dkk-1 promoter showed that one putative glucocorticoid responsive element-like sequence located from -788 to -774bp is essential for the enhancement of the Dkk-1 promoter activity by dexamethasone in human osteoblasts. Since the Wnt signal is now recognized as a crucial regulator for bone formation, the Dkk-1 enhanced by glucocorticoid may inhibit the Wnt signal in osteoblasts, which may be involved in the pathogenesis of glucocorticoid-induced osteoporosis. PMID- 15110783 TI - Atorvastatin reduces CD68, FABP4, and HBP expression in oxLDL-treated human macrophages. AB - With the aim of identifying new target genes that could contribute to limit foam cell formation, we analyzed changes in the pattern of gene expression in human THP-1 macrophages treated with atorvastatin and oxidized-LDL (oxLDL). To this end, we used a human cDNA array containing 588 cardiovascular-related cDNAs. Exposure to oxLDL resulted in differential expression of 26 genes, while coincubation with atorvastatin modified the expression of 29 genes, compared to treatment with oxLDL alone. Changes in the expression of candidate genes, potentially connected to the atherosclerotic process, were confirmed by quantitative RT-PCR and Western blot. We show that atorvastatin prevents the increase in the expression of scavenger receptor CD68 and that of fatty acid binding protein 4 caused by oxLDL. In addition, atorvastatin reduces the expression of HDL-binding protein, apolipoprotein E, and matrix metalloproteinase 9. These findings are relevant to understand the direct antiatherogenic effects of statins on macrophages. PMID- 15110784 TI - Anti-HCV activities of selective polyunsaturated fatty acids. AB - HCV infection can lead to chronic infectious hepatitis disease with serious sequelae. Interferon-alpha, or its PEGylated form, plus ribavirin is the only treatment option to combat HCV. Alternative and more effective therapy is needed due to the severe side effects and unsatisfactory curing rate of the current therapy. In this study, we found that several polyunsaturated fatty acids (PUFAs) including arachidonic acid (AA), docosahexaenoic acid (DHA), and eicosapentaenoic acid (EPA) are able to exert anti-HCV activities using an HCV subgenomic RNA replicon system. The EC(50) (50% effective concentration to inhibit HCV replication) of AA was 4microM that falls in the range of physiologically relevant concentration. At 100microM, alpha-linolenic acid, gamma-linolenic, and linoleic acid only reduced HCV RNA levels slightly and saturated fatty acids including oleic acid, myristic acid, palmitic acid, and steric acid had no inhibitory activities toward HCV replication. When AA was combined with IFN alpha, strong synergistic anti-HCV effect was observed as revealed by an isobologram analysis. It will be important to determine whether PUFAs can provide synergistic antiviral effects when given as food supplements during IFN-based anti-HCV therapy. Further elucidation of the exact anti-HCV mechanism caused by AA, DHA, and EPA may lead to the development of agents with potent activity against HCV or related viruses. PMID- 15110785 TI - Mineralocorticoid receptor antagonists do not block rapid ERK activation by aldosterone. AB - Aldosterone can elicit rapid nongenomic effects both in vivo and in vitro, often mediated by signal transduction cascades. However, it is not understood how these rapid effects are initiated. In this study we show that aldosterone leads to rapid activation of mitogen activated protein kinases ERK1/2 in the cortical collecting duct cell line M-1. Inhibitors of transcription and translation could not block this activation, which suggests an extranuclear (nongenomic) mechanism. Although it is known that M-1 cells do not contain a transcriptionally functional MR, it is not known whether a closely related protein still could mediate the effects, or an unrelated nonclassic receptor. To test this hypothesis, the effects of four classical mineralocorticoid receptor antagonists were studied. None of the compounds could block the response to aldosterone. Altogether, the data suggest that rapid aldosterone effects in M-1 cells are initiated by a receptor different from the classical mineralocorticoid receptor. PMID- 15110786 TI - Proteomic analysis of the TGF-beta signaling pathway in pancreatic carcinoma cells using stable RNA interference to silence Smad4 expression. AB - Smad4 is a tumor-suppressor gene that is lost or mutated in 50% of pancreatic carcinomas. Smad4 is also an intracellular transmitter of transforming growth factor-beta (TGF-beta) signals. Although its tumor-suppressor function is presumed to reside in its capacity to mediate TGF-beta-induced growth inhibition, there seems to be a Smad4-independent TGF-beta signaling pathway. Here, we succeeded in establishing Smad4 knockdown (S4KD) pancreatic cancer cell lines using stable RNA interference. Smad4 protein expression and TGF-beta-Smad4 signaling were impaired in S4KD cells, and we compared the proteomic changes with TGF-beta stimulation using two-dimensional gel electrophoresis (2-DE) and mass spectrometry. We identified five proteins that were up-regulated and seven proteins that were down-regulated; 10 of them were novel targets for TGF-beta. These proteins function in processes such as cytoskeletal regulation, cell cycle, and oxidative stress. Introducing siRNA-mediated gene silencing into proteomics revealed a novel TGF-beta signal pathway that did not involve Smad4. PMID- 15110787 TI - Mismatch repair-mediated G2/M arrest by 6-thioguanine involves the ATR-Chk1 pathway. AB - DNA mismatch repair (MMR) deficiency in human cancers is associated with resistance to a spectrum of clinically active chemotherapy drugs, including 6 thioguanine (6-TG). We and others have shown that 6-TG-induced DNA mismatches result in a prolonged G2/M cell cycle arrest followed by apoptosis in MMR(+) human cancer cells, although the signaling pathways are not clearly understood. In this study, we found that prolonged (up to 4 days) treatment with 6-TG (3microM) resulted in a progressive phosphorylation of Chk1 and Chk2 in MMR(+) HeLa cells, correlating temporally with a drug-induced G2/M arrest. Transfection of HeLa cells with small interfering RNA (siRNA) against the ataxia telangiectasia-related (ATR) kinase or against the Chk1 kinase destroyed the G2/M checkpoint and enhanced the apoptosis following 6-TG treatment. On the other hand, the induction of a G2/M population by 6-TG was similar in ATM(-/-) and ATM(+) human fibroblasts, suggesting that the ATM-Chk2 pathway does not play a major role in this 6-TG response. Our results indicate that 6-TG DNA mismatches activate the ATR-Chk1 pathway in the MMR(+) cells, resulting in a G2/M checkpoint response PMID- 15110788 TI - The gene-silencing efficiency of siRNA is strongly dependent on the local structure of mRNA at the targeted region. AB - The gene-silencing effect of short interfering RNA (siRNA) is known to vary strongly with the targeted position of the mRNA. A number of hypotheses have been suggested to explain this phenomenon. We would like to test if this positional effect is mainly due to the secondary structure of the mRNA at the target site. We proposed that this structural factor can be characterized by a single parameter called "the hydrogen bond (H-b) index," which represents the average number of hydrogen bonds formed between nucleotides in the target region and the rest of the mRNA. This index can be determined using a computational approach. We tested the correlation between the H-b index and the gene-silencing effects on three genes (Bcl-2, hTF, and cyclin B1) using a variety of siRNAs. We found that the gene-silencing effect is inversely dependent on the H-b index, indicating that the local mRNA structure at the targeted site is the main cause of the positional effect. Based on this finding, we suggest that the H-b index can be a useful guideline for future siRNA design. PMID- 15110789 TI - Modulation of cell proliferation and gene expression by alpha-tocopheryl phosphates: relevance to atherosclerosis and inflammation. AB - The effect of a mixture of alpha-tocopheryl phosphate and di-alpha-tocopheryl phosphate (TPm) was studied in vitro on two cell lines, RASMC (from rat aortic smooth muscle) and human THP-1 monocytic leukaemia cells. Inhibition of cell proliferation by TPm was shown in both lines and occurred with TPm at concentrations lower than those at which alpha-tocopherol was equally inhibitory. TPm led in non-stimulated THP-1 cells to inhibition of CD36 mRNA and protein expression, to inhibition of oxidized low density lipoprotein surface binding and oxLDL uptake. In non-stimulated THP-1 cells, alpha-tocopherol had only very weak effects on these events. Contrary to alpha-tocopherol, TPm was cytotoxic to THP-1 cells at high concentrations. Thus, TPm is able to inhibit the major aggravating elements involved in the progression of atherosclerosis. The higher potency of TPm may be due to a better uptake of the molecule and to its intracellular hydrolysis, providing more alpha-tocopherol to sensitive sites. Alternatively, a direct effect of the phosphate ester on specific cell targets may be considered. PMID- 15110790 TI - Ectopic expression of the maize chromosomal HMGB1 protein causes defects in root development of tobacco seedlings. AB - Chromatin-associated high mobility group (HMG) proteins of the HMGB family are versatile architectural factors assisting various DNA-dependent processes such as transcription and recombination. Here, transgenic tobacco lines were generated that ectopically express the maize HMGB1 protein, as detected by immunoblot analyses. The shoot morphology of HMGB1 expressing plants does not differ from that of control plants. By contrast, tobacco seedlings expressing HMGB1 are impaired in the growth of the primary root, relative to control plants. The reduced primary root length is correlated with the accumulation of small cells in the cell division zone (but not in the elongation zone) of the roots of transgenic plants. This "short-root" phenotype is specific for HMGB1, as is not observed with HMGB4 expressing plants, and it is transient in that it is restricted to young seedlings (40 tryptamine-related compounds. No tryptamine analog was found to bind with substantially higher affinity than 5-HT. The results indicate that hydrogen bonding plays a key role in the 5-HT1E/receptor interaction. This finding was supported using quantitative structure-activity analysis (QSAR) techniques such as comparative molecular field analysis (CoMFA) and the program QsarIS. PMID- 15110838 TI - ERbeta ligands. Part 2: Synthesis and structure-activity relationships of a series of 4-hydroxy-biphenyl-carbaldehyde oxime derivatives. AB - A series of biphenyl carbaldehyde oximes (6) was prepared and shown to have significant selectivity for the estrogen receptor-beta (ERbeta). The exploitation of the oxime moiety as a hydrogen bond donating group, which mimicked the C-ring of genistein makes these compounds unique. Molecular modeling studies showed the oxime moiety hydrogen bonding to the histidine residue, which was supported by the structure-activity relationships. The most potent compounds in this study had IC50 values in a radioligand binding assay of between 8-35 nM. Among the most selective compounds were 6i and 6s (49- and 31-fold ERbeta selective, respectively). PMID- 15110839 TI - S1 subsite in snake venom thrombin-like enzymes: can S1 subsite lipophilicity be used to sort binding affinities of trypsin-like enzymes to small-molecule inhibitors? AB - Thrombin-like enzymes isolated from snake venoms comprise a group of serine proteinases responsible for many important coagulation disorders in the envenomed victims. Besides, these proteinases have great biotechnological interest as antithrombotic agents and as diagnostic tools. However, in spite of the recent overflow of snake venom thrombin-like enzymes (SVTLEs) on protein sequence databases, there is a lack of three-dimensional (3D) structural information on this family. Without such 3D structures available many aspects of the biological function and biochemical properties of these enzymes still remain obscure. Therefore, we have gone through a series of computational techniques, which enabled us to identify the set of residues involved in molecular recognition of inhibitors bound to the S1 subsite of snake venom thrombin-like enzymes (SVTLEs) and ultimately conclude that nonpolar (van der Waals) intermolecular interactions and ligand's hydrophobicity are the most important factors affecting binding affinities to the S1 subsite of a SVTLE isolated from the venom of Lachesis muta muta (Lmm-TLE). Consequently, we have proposed that S1 subsite lipophilicity may be used to sort binding affinities of trypsin-like enzymes to small molecules by showing that the inhibitory potency of several S1-directed compounds follows subsite lipophilicity among Lmm-TLE and other three homologous proteases. Noteworthy, in the course of our analyses we determined that thrombin's S1 subsite should, in fact, be considered less lipophilic than that of trypsin if we account for the presence of the sodium-controlled water channel communicating with the S1 subsite in the coagulant enzyme. PMID- 15110840 TI - Comparison of the dark and light-induced toxicity of thio and seleno analogues of the thiopyrylium dye AA1. AB - 2,6-Bis(4-anilino)-4-(4-N,N-dimethylanilino)thiopyrylium chloride (AA1) and selenopyrylium chloride (AA1-Se) and 2,6-bis(4-anilino)-4-(4-N morpholinophenyl)thiopyrylium chloride (1) and -selenopyrylium chloride (2) were prepared via the addition of 4-N,N-dimethylanilino magnesium bromide and 4-N morpholinophenyl magnesium bromide to chalcogenopyranones 3 followed by treatment with HCl gas then water. Cellular uptake of these dyes varied from 12+/ 3fmol/cell for AA1 to 150+/-40 fmol/cell for AA1-Se. upon exposure to 5 x 10(-5) M solutions of the dyes for 3 h. Exposure of cell cultures to 1.8 J/cm2) of 360 750-nm light following incubation with 1 x 10(-6) M of either AA1, 1, or 2 for 24h resulted in no significant additional phototoxicity while AA1-Se showed a significant (p < 0.05) reduction in cell viability from 81% to 46%. Thiopyrylium dyes AA1 and 1 showed significant dark toxicity relative to selenopyrylium dyes AA1-Se and 2, respectively. AA1 was the only one of the four dyes to show inhibition of whole-cell mitochondrial cytochrome c oxidase activity in the dark. Irradiation of whole cells or mitochondrial suspensions treated with AA1, AA1-Se, or 2 gave inhibition of mitochondrial cytochrome c oxidase activity. Studies of JC-1-efflux indicated that all four cationic dyes accelerated the loss of JC-1 from the mitochondria, which suggests that all four dyes target the mitochondria. PMID- 15110841 TI - Taxol derivatives are selective inhibitors of DNA polymerase alpha. AB - During screening for mammalian DNA polymerase inhibitors, we found and succeeded in isolating a potent inhibitor from a higher plant, Taxus cuspidate. The compound was unexpectedly determined to be taxinine, an intermediate of paclitaxel (taxol) metabolism. Taxinine was found to selectively inhibit DNA polymerase alpha (pol.alpha) and beta (pol.beta). We therefore, tested taxol and other derivatives and found that taxol itself had no such inhibitory effect, and only taxinine could inhibit both pol.alpha and beta. The other compounds used, one derivative, cephalomannine, and five intermediates synthesized chemically inhibited only the pol.alpha activity in vitro. None of the compounds, including taxinine, influenced the activities of the other DNA polymerases, which are reportedly targeted by many pol.beta inhibitors. With both pol.alpha and beta, all of the compounds tested noncompetitively inhibited with respect to both the DNA template-primer and the dNTP substrate. PMID- 15110842 TI - Cinnamic amides of (S)-2-(aminomethyl)pyrrolidines are potent H3 antagonists. AB - New imidazole-free H3 antagonists have been found in a series of cinnamic amides of (S)-(aminomethyl)pyrrolidines. The influence of the substituent on the aromatic moiety on the potency and the inhibition of three cytochrome P450 subtypes are also described. PMID- 15110843 TI - Synthesis and activity of 1H-benzimidazole and 1H-benzotriazole derivatives as inhibitors of Acanthamoeba castellanii. AB - Chloro-, bromo- and methyl- analogues of 1H-benzimidazole and 1H-benzotriazole and their N-alkyl derivatives have been synthesized and tested in vitro against the protozoa Acanthamoeba castellanii. The results indicate that 5,6-dimethyl-1H benzotriazole (11) and 5,6-dibromo-1H-benzotriazole (14) have higher efficacy than the antiprotozoal agent chlorohexidine. PMID- 15110844 TI - Clozapine derived 2,3-dihydro-1H-1,4- and 1,5-benzodiazepines with D4 receptor selectivity: synthesis and biological testing. AB - Novel 4-arylpiperazin-1-yl-substituted 2,3-dihydro-1H-1,4- and 1H-1,5 benzodiazepines and their aza-analogues were synthesized as debenzoclozapine derivatives for evaluation as potential D4-ligands. While Ki values of some of the title compounds came within the range of clozapine, they showed an impressively greater selectivity over other dopamine receptor subtypes, especially D2. For the most promising compounds, intrinsic activity and binding properties to serotonin 5-HT1A and 5-HT2 were also determined. PMID- 15110845 TI - 3D-QSAR analysis of conformationally constrained diacylglycerol (DAG) analogues as potent protein kinase C (PK-C) ligands. AB - A study of the quantitative structure activity relationships (QSARs) was performed based on the binding affinity (pKi) values of 32 protein kinase C (PK C) ligands. The QSAR study was carried out by using both three-dimensional descriptors (the steric and electrostatic CoMFA fields) and the physicochemical properties (logP values). The CoMFA analysis provided a reasonable QSAR model, with a cross-validated q2 value of 0.671 and a conventional r2 value of 0.956, which was confirmed by the satisfactory prediction of the experimental binding affinity (pKi) values for a series of 3-alkylidene-5,5-disubstituted tetrahydro-2 furanones included in the test set. The resultant QSAR model will be useful for designing highly potent and selective PK-C ligands. PMID- 15110846 TI - Fluorinated phenylcyclopropylamines. Part 3: Inhibition of monoamine oxidase A and B. AB - Fluorinated phenylcyclopropylamines and alkylamines were examined as inhibitors of recombinant human liver monoamine oxidase A (MAO A) and B (MAO B). For a series of trans- and cis-2-fluoro-2-phenylcyclopropylamine analogues, the presence of fluorine attached to a cyclopropane ring was found to result in an increase in inhibitory activity towards both MAO A and B. In addition, p substitution of electron-withdrawing groups such as Cl and F in the aromatic ring of the trans-isomers increased the inhibition of both enzymes. (1S,2S)-2-Fluoro-2 phenylcyclopropylamine was a more potent inhibitor of both MAO A and B than was the (1R,2R)-enantiomer, indicating that the presence of fluorine has no influence on the enantioselectivity of MAO inhibition, since a similar effect of stereochemistry has been reported for tranylcypromine. Interestingly, fluorination at the 2-position of 1-phenycyclopropylamine, which is known as a selective inhibitor of MAO B relative to MAO A, reversed the selectivity and resulted in a potent inhibitor selective for MAO A. All inhibitors showed time- and concentration-dependent inhibition for both enzymes, with the exception of trans-2-fluoro-2-phenylcyclopropyl ethylamine, which acts as a competitive and reversible MAO A selective inhibitor. PMID- 15110847 TI - Naphtho[2,1-b][1,5] and [1,2-f][1,4]oxazocines as selective NK1 antagonists. AB - Previously we reported on the synthesis and properties of a series of highly potent piperidinyl 2-subsituted-3-cyano-1-naphthamide NK1 antagonists that includes 3 and 4. Here we report our efforts to alleviate a troublesome atropisomeric property of those derivatives by introduction of a tethering bridge that, in addition, could be used to lock the resulting cyclic derivatives in a purported NK1 pharmacophore conformation. Using 3 as a starting point, the naphtho[2,1-b][1,5]oxazocine, 17, was found to contain the optimal ring tether size (8) for retaining NK1 activity, was more NK1 versus NK2 selective, and reduced the number of atropisomers from four to two. Cyclic derivatives 29 and 32, which exist as essentially single atropisomers in the purported pharmacophore conformation, were prepared in the closely related naphtho[1,2-f][1,4]oxazocine series as part of an effort to use mono methyl substitution of the tethering bridge as a conformation stabilizing factor. Both 29 and 32 were found to be less active as NK1 antagonists than the non-methylated parent 28 possibly due to methyl group destabilization of receptor interaction. We discuss the above findings in the context of a previously proposed NK1 pharmacophore model and present a further refinement of that model. PMID- 15110848 TI - A predictive pharmacophore model of human melanocortin-4 receptor as derived from the solution structures of cyclic peptides. AB - Using nuclear magnetic resonance (NMR) spectroscopy, we have determined the solution structures for a series of potent agonists for the human melanocortin-4 receptor (hMC4R), based on the cyclic peptide MT-II [Ac-Nle-cyclo-(Asp-Lys) (Asp His-(D)Phe-Arg-Trp-Lys)-NH2]. Members of this series were designed to improve selectivity for MC4R versus the other melanocortin receptors, and to reduce the flexibility of the side chains. The most selective and rigid analog [penta cyclo(D-K)-Asp-Apc-(D)Phe-Arg-(2S,3S)-beta-methylTrp-Lys-NH2] was found to be a full agonist of hMC4R with an EC50 of 11nM against hMC4R, and to exhibit 65-fold selectivity against hMC1R. This compound represents the most constrained hMC4R peptide agonist described to date. A beta-turn structure was conserved among all of the cyclic peptides studied. The rigidity of the analogs allowed an exceptionally well-defined pharmacophore model to be derived. This model was used to perform a virtual screen using a library of 1000 drug-like compounds, to which a small set of known potent ligands had been intentionally added. The utility of the model was validated by its ability to identify the known ligands from among this large library. PMID- 15110849 TI - Differential effects of synthesized 2'-oxygenated chalcone derivatives: modulation of human cell cycle phase distribution. AB - Ten structurally related 2'-oxygenated chalcone derivatives, bearing either hydroxy and/or methoxy substituents on the A and B rings, were synthesized through Claisen-Schmidt condensation. The synthesis procedure was relatively easy and had an acceptable yield. The in vitro cytotoxicities of these compounds against the human tumor cells such as Jurkat, U937 cells, and normal cells PHA stimulated PBMCs were investigated. Among those, compounds 1 (IC50 = 2.5 microM), 2 (1.7 microM), and 8 (3.2 microM) showed potent inhibitory activity toward Jurkat cell line. In parallel, compounds 1 (6.7 microM), 2 (1.5 microM), and 10 (5.3 microM) showed the highest activity against U937 cell line. However, the chalcones also inhibit the PHA stimulated PBMCs cells, but the IC50 values were relatively high when compared to the tumor cell line values. Studies were also on the effect of synthesized chalcones on the cell cycle phase distribution. In Jurkat cell line, compounds 7 and 9 showed the highest activity and the most striking effect in reduction of the percentage of cells in the S phase, which was associated with an increase of cells in G2/M phase. In U937 cell line, compound 3 increased the proportion of cells in the G0/G1 phase and reduced the proportion in S phase. In contrast, compounds 1, 9, and 10 showed a decrease effect on the percentage of cells in S phase and an increase effect on the percentage of cells in the G2/M phase of the cell cycle. Whereas in the case of PHA stimulated PBMCs, compounds 1, 4, 8, and 10 increased the percentage of cells in G2/M phase, which was associated with a decrease effect in the S phase of the cell cycle. PMID- 15110850 TI - An investigation of the N-demethylation of 3-deoxymorphine and the affinity of the alkylation products to mu, delta, and kappa receptors. AB - The N-demethylation of 3-deoxymorphine (1) was investigated using methyl chloroformate and hydrazine. 3-Deoxynormorphine (2) was obtained in 70% yield, and 3-deoxydihydronormorphine (3) was also obtained as a side product. The mu, delta, and kappa receptor binding affinity of a series of N-substituted 3 deoxynormorphines 6 and 7 and N-substituted 3-deoxydihydronormorphines 8-11 was also determined. PMID- 15110851 TI - Synthesis and structure activity relationship of guanidines as NPY Y5 antagonists. AB - A series of bis-aryl substituted guanidines have been discovered as potent NPY Y5 antagonists. The SAR and in vitro metabolic stability of these compounds are discussed. PMID- 15110852 TI - 3D-QSAR studies of pyruvate dehydrogenase kinase inhibitors based on a divide and conquer strategy. AB - PDHK is a highly specific enzyme, which inhibits PDC thereby reducing the conversion of pyruvate to AcetylCoA leading to increased glucose and lactate level contributing to various pathological disease states. 3D-QSAR CoMFA studies were performed on diverse PDHK inhibitors based on maximum common substructural alignments of different classes of molecules with the selected reference molecule using a divide and conquer strategy. Statistically robust CoMFA model was obtained with a cross-validated correlation coefficient of 0.561 and conventional correlation coefficient of 0.990. Predictive correlation coefficient r2(pred) was found to be 0.875. PMID- 15110853 TI - Carbonic anhydrase inhibitors: aromatic and heterocyclic sulfonamides incorporating adamantyl moieties with strong anticonvulsant activity. AB - A series of aromatic/heterocyclic sulfonamides incorporating adamantyl moieties were prepared by reaction of aromatic/heterocyclic aminosulfonamides with the acyl chlorides derived from adamantyl-1-carboxylic acid and 1-adamantyl-acetic acid. Related derivatives were obtained from the above-mentioned aminosulfonamides with adamantyl isocyanate and adamantyl isothiocyanate, respectively. Some of these derivatives showed good inhibitory potency against two human CA isozymes involved in important physiological processes, CA I, and CA II, of the same order of magnitude as the clinically used drugs acetazolamide and methazolamide. The lipophilicity of the best CA inhibitors was determined and expressed as their experimental log k' IAM and theoretical ClogP value. Their lipophilicity was propitious with the crossing of the blood-brain barrier (log k' > IAM > 1.35). The anticonvulsant activity of some of the best CA inhibitors reported here has been evaluated in a MES test in mice. After intraperitoneal injection (30 mg kg(-1)), compounds A8 and A9 exhibited a high protection against electrically induced convulsions (> 90%). Their ED50 was 3.5 and 2.6 mg kg(-1), respectively. PMID- 15110854 TI - Structural variations of 1-(4-(phenoxymethyl)benzyl)piperidines as nonimidazole histamine H3 receptor antagonists. AB - Recent bioisoteric replacements in histamine H3 receptor ligands with an exchange of the imidazole moiety by a piperidino group as well as of the trimethylene chain in 4-((3-phenoxy)propyl)-lH-imidazole derivatives (proxifan class) by an alpha,alpha'-xylendiyl linker represents the starting point in the development of 1-(4-(phenoxymethyl)benzyl)piperidines as a new class of nonimidazole histamine H3 receptor antagonists. According to different strategies in optimization of imidazole-containing antagonists the central benzyl phenyl ether moiety was replaced by numerous other polar functionalities. Additionally, the ortho- and meta-analogues of the lead were synthesized to determine the influence of the position of the piperidinomethyl substituent. The new compounds were tested in an in vitro binding assay for their affinities for cloned human H3 receptors stably expressed in CHO-K1 cells and for their oral in vivo potencies brain in a functional screening assay in the brain of mice. Additionally, activities of selected compounds were determined in the guinea-pig ileum functional test model. In contrast to the analogues ortho-substituted compounds all other compounds maintained respectable affinities for the human H3 receptor (-log Ki values 6.3 7.5). Despite the results from other classes of compounds the 4-methyl substituted derivatives generally displayed higher affinities than the corresponding 4-chloro substituted compounds. In vivo only the inverse phenyl benzyl ether (3) showed worthwhile antagonist potencies. PMID- 15110855 TI - Synthesis and pharmacological properties of benzamide derivatives as selective serotonin 4 receptor agonists. AB - A series of 4-amino-5-chloro-2-methoxy-N-(piperidin-4-ylmethyl)benzamides with a polar substituent group at the 1-position of the piperidine ring was synthesized and evaluated for its effect on gastrointestinal motility. The benzoyl, phenylsulfonyl, and benzylsulfonyl derivatives accelerated gastric emptying and increased the frequency of defecation. One of them, 4-amino-N-[1-[3 (benzylsulfonyl)propyl]piperidin-4-ylmethyl]-5-chloro-2-methoxybenzamide (13a, Y 36912), was a selective 5-HT4 receptor agonist offering potential as a novel prokinetic with reduced side effects derived from 5-HT3- and dopamine D2 receptor binding affinity. In the oral route of administration, this compound enhanced gastric emptying and defecation in mice, and has a possibility as a prokinetic agent, which is effective on both the upper and the lower gastrointestinal tract. PMID- 15110856 TI - An o-nitrobenzyl scaffold for peptide ligation: synthesis and applications. AB - Chemical ligation approaches facilitate the chemoselective assembly of unprotected peptides in aqueous solution. Here, two photolabile auxiliaries are described that enlarge the applicability of native chemical ligation to non cysteine targets. The auxiliaries, designed to allow reaction with thioester peptides, generate an amide bond between the two initial fragments. The o nitrobenzyl tertiary benzylamide that is formed at the ligation junction can be transformed into a native amide group under mild photolysis conditions. The veratryl auxiliary was found to be excessively labile during peptide purification and ligation. However, the auxiliary based on the o-nitrobenzyl group shows all the necessary properties for a general application in routine peptide and protein synthesis. In addition, the auxiliary linked to the N-terminus can be efficiently photolyzed, suggesting a new approach for the generation of photocaged amines. Synthesis, solid phase introduction onto peptide chains, ligation properties and photolysis in water are described, and a careful study of compatibility of the method with potentially fragile peptide side chains is reported. PMID- 15110857 TI - Synthesis, in vitro and in vivo activity of benzophenone-based inhibitors of steroid sulfatase. AB - Steroid sulfatase (STS) is an important new therapeutic target in oncology. Attempts to design nonsteroidal STS inhibitors, because of the oestrogenicity of the original lead oestrone 3-O-sulfamate in rodents, have led to the discovery of benzophenone-4,4'-O,O-bis-sulfamate (BENZOMATE, 3). The nonfused bicyclic BENZOMATE is a highly potent STS inhibitor in vitro, inhibiting STS activity in intact MCF-7 breast cancer cells by > 70% at 0.1 microM and in placental microsomes by > 98% at 10 microM. When MCF-7 cells were pre-treated with 3 at 1 microM and then washed to remove unbound inhibitor, the initial 94% inhibition was reduced to 89% suggesting that 3, like other sulfamate-based STS inhibitors, inhibits the enzyme irreversibly. This agent also inhibits rat liver STS activity by 84% and 93% respectively 24 h after a single dose of 1 or 10 mg/kg, demonstrating that BENZOMATE possesses similar in vivo potency to the established potent nonsteroidal inhibitor 667COUMATE. Several modifications were made to BENZOMATE structurally and effects on in vitro activity were examined. These structure-activity relationship studies show that its carbonyl and bis-sulfamate groups are pivotal for activity, although conformational flexibility is not required. Two rigid anthraquinone-based sulfamate derivatives however showed inhibitory activity significantly better than BENZOMATE in the MCF-7 cell assay. BENZOMATE and related analogues therefore represent an important class of non steroidal STS inhibitor and lead compounds for future drug design. PMID- 15110858 TI - Binding of an influenza A virus to a neomembrane measured by surface plasmon resonance. AB - Neomembranes composed of either bovine brain lipid that contains sialoglycolipids or egg yolk lecithin that does not, were formed on an HPA sensor chip and used to study the binding of influenza A virus in real time by surface plasmon resonance. Virus bound only to the bovine brain lipid membrane. This was confirmed by an 84% reduction in virus binding after treatment of the neomembrane with neuraminidase. Binding was temperature dependent, being highest at 30-35 degrees C and lower at 10 degrees C. Surprisingly, the rate of complex formation was enhanced, rather than inhibited, by the presence of 1.34-25.2 x 10(6) molecules of free NANA per virus binding site and the rate of dissociation was lower suggesting that the complex was more stable. The free energy of association to form the transition complex was increased by 3 kJ mol(-1) and there was an almost 10-fold increase in the enthalpy of complex formation in the presence of free NANA. These results show the value of surface plasmon resonance for measuring complex molecular interactions in real time, and provide a model that can be used to study the effectiveness of inhibitors of attachment of influenza virus to its receptor molecules. PMID- 15110859 TI - Purification and partial amino acid sequences of the enzyme vinorine synthase involved in a crucial step of ajmaline biosynthesis. AB - The acetyl-CoA-dependent enzyme vinorine synthase was isolated from hybrid cell suspension cultures of Rauvolfia serpentina and Rhazya stricta. The sarpagan-type alkaloid gardneral was used as a substrate of the enzyme leading to the ajmalan type 10-methoxyvinorine. An HPLC-based assay was developed to monitor vinorine synthase activity, which allowed establishing a five step purification procedure combining anion exchange, hydrophobic interaction, hydroxyapatite and gel filtration. Purification resulted in a yield of 0.2% and an approximately 991 fold enrichment of the acetyltransfer activity. SDS-PAGE analysis showed a Mr for the enzyme of approximately 50 kDa. The four peptide fragments generated by proteolysis of the pure enzyme with endoproteinase LysC and the N-terminal part of the enzyme were sequenced. The enzyme preparation (> 875-fold enrichment) delivering the N-terminal sequence was isolated from R. serpentina cell suspensions. Sequence alignment of the five peptides showed highest homologies in a range of 30-71% to acetyltransferases from other higher plants involved in natural plant product biosynthesis. Based on the partial sequences vinorine synthase is probably a novel member of the BAHD enzyme super family. PMID- 15110860 TI - Acetyltransfer in natural product biosynthesis--functional cloning and molecular analysis of vinorine synthase. AB - Vinorine synthase (EC 2.3.1.160) catalyses the acetyl-CoA- or CoA-dependent reversible formation of the alkaloids vinorine (or 11-methoxy-vinorine) and 16 epi-vellosimine (or gardneral). The forward reaction leads to vinorine, which is a direct biosynthetic precursor along the complex pathway to the monoterpenoid indole alkaloid ajmaline, an antiarrhythmic drug from the Indian medicinal plant Rauvolfia serpentina. Based on partial peptide sequences a cDNA clone was isolated and functionally expressed in Escherichia coli. The Km values of the native enzyme for gardneral and acetyl-CoA were determined to be 7.5 and 57 microM. The amino acid sequence of vinorine synthase has highest level of identity (28-31%) to that of Papaver salutaridinol acetyltransferase, Fragaria alcohol acyltransferase, and Catharanthus deacetylvindoline acetyltransferase involved in morphine, flavor, and vindoline biosynthesis, respectively. Vinorine synthase is a novel member of the BAHD superfamily of acyltransferases. Site directed mutagenesis of 13 amino acid residues provided clear evidence that both, His160 and Asp164 of the consensus sequence HxxxD belong to the catalytic center. The mutations also showed that an amino acid triad is not characteristic of vinorine synthase. The experiments demonstrated the importance of the conserved motif SxL/I/VD near the N-terminus and the consensus sequence DFGWG near the C terminal. PMID- 15110861 TI - 3D-QSAR CoMFA/CoMSIA studies on Urokinase plasminogen activator (uPA) inhibitors: a strategic design in novel anticancer agents. AB - Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) was performed on a series of indole/benzoimidazole-5 carboxamidines as urokinase plasminogen activator (uPA) inhibitors. The ligand molecular superimposition on template structure was performed by atom/shape-based RMS fit, multifit, and RMSD fit methods. The removal of two outliers from the initial training set of 30 molecules improved the predictivity of the models. The statistically significant model was established from 28 molecules, which were validated by evaluation of test set of nine compounds. The atom-based RMS alignment yielded best predictive CoMFA model (r2(cv) = 0.611, r2(cnv) = 0.778, F value = 43.825, r2(bs) = 0.842, r2(pred) = 0.616 with two components) while the CoMSIA model yielded (r2(cv) = 0.499, r2(cnv) = 0.976, F value=96.36, r2(bs) = 0.993, r2(pred) = 0.694 with eight components). The contour maps obtained from 3D QSAR studies were appraised for the activity trends of the molecules analyzed. The results indicate that the steric, electrostatic, and hydrogen bond donor/acceptor substituents play significant role in uPA activity and selectivity of these compounds. The data generated from the present study can be used as putative pharmacophore in the design of novel, potent, and selective urokinase plasminogen activator inhibitors as cancer therapeutics. PMID- 15110862 TI - Honey bee (Apis mellifera) transferrin-gene structure and the role of ecdysteroids in the developmental regulation of its expression. AB - Social life is prone to invasion by microorganisms, and binding of ferric ions by transferrin is an efficient strategy to restrict their access to iron. In this study, we isolated cDNA and genomic clones encoding an Apis mellifera transferrin (AmTRF) gene. It has an open reading frame (ORF) of 2136 bp spread over nine exons. The deduced protein sequence comprises 686 amino acid residues plus a 26 residues signal sequence, giving a predicted molecular mass of 76 kDa. Comparison of the deduced AmTRF amino acid sequence with known insect transferrins revealed significant similarity extending over the entire sequence. It clusters with monoferric transferrins, with which it shares putative iron-binding residues in the N-terminal lobe. In a functional analysis of AmTRF expression in honey bee development, we monitored its expression profile in the larval and pupal stages. The negative regulation of AmTRF by ecdysteroids deduced from the developmental expression profile was confirmed by experimental treatment of spinning-stage honey bee larvae with 20-hydroxyecdysone, and of fourth instar-larvae with juvenile hormone. A juvenile hormone application to spinning-stage larvae, in contrast, had only a minor effect on AmTRF transcript levels. This is the first study implicating ecdysteroids in the developmental regulation of transferrin expression in an insect species. PMID- 15110863 TI - Proteomic analysis of brown planthopper: application to the study of carbamate toxicity. AB - Toxicity to o-sec-butylphenyl methylcarbamate compound (BPMC) was analyzed in the rice brown planthopper, Nilaparvata lugens, using a differential proteomics approach of identifying proteins on two dimensional-polyacrylamide gel electrophoresis (2D-PAGE). Proteome analysis from BPMC-treated brown planthopper resulted in the modulation of 22 proteins at the expression level as compared to control samples on coomassie brilliant blue (CBB) stained gels. Out of total 22 proteins, 10 proteins showed elevated expression, eight proteins showed decreased expression and four proteins showed specific expression after insecticide treatment. The N-terminal sequences of seven out of 22 proteins were determined by a gas-phase protein sequencer. The internal amino acid sequences of the 15 proteins were determined by the sequence analyses of peptides obtained by Cleveland peptide mapping method and were compared with those of the known proteins available in public databases and the EST database of the brown planthopper in our laboratory to understand the nature of the proteins. Sequence analyses revealed that the expression of putative serine/threonine protein kinase, paramyosin, HSP 90, beta-tubulin, calreticulin, ATP synthase, actin and tropomyosin was elevated, and that of beta-mitochondrial processing peptidase, dihydrolipoamide dehydrogenase, enolase and acyl-coA dehydrogenase was reduced due to the exposure of BPMC. The differential expression of these proteins reflects the overall change in cellular structure and metabolism after insecticide treatment. PMID- 15110864 TI - Molecular comparisons of the Culex pipiens (L.) complex esterase gene amplicons. AB - The amplification of carboxylesterase genes is a mechanism of organophosphate resistance in Culex mosquitoes. Amplified carboxylesterase genes from an insecticide resistant Culex pipiens strain collected in Cyprus were analysed and compared to other Culex amplified carboxylesterase alleles. A 12 kb section of genomic DNA containing two gene loci coding for carboxylesterase alleles A5 and B5 was cloned and sequenced. A comparison between this amplicon and one from a strain with co-amplified carboxylesterase alleles A2 and B2 revealed a number of differences. The intergenic spacer was 3.7 kb in length in the A5-B5 amplicon (2.7 kb in A2-B2) and contained putative Juan and transposable elements upstream of B5. A fragment of a gene with high homology to aldehyde oxidase was also present immediately downstream of A5. The comparison revealed no differences that would explain the successful spread of the A2-B2 amplicon worldwide whilst the A5 B5 amplicon is restricted to the Mediterranean. PMID- 15110865 TI - Proteolytic activity of Boophilus microplus Yolk pro-Cathepsin D (BYC) is coincident with cortical acidification during embryogenesis. AB - In a previous report (Parasitology 116 (1998) 525) we isolated and characterized Boophilus Yolk pro-Cathepsin (BYC), an aspartic proteinase precursor from the eggs of the hard tick. The present study was designed to characterize the function of BYC in the consumption of vitellin (VT), the major yolk protein, during embryogenesis. Both purified BYC and total egg homogenate proteolytic activity showed a similar pH dependence profile with an acidic optimum. Purified BYC presented higher activity against VT as a substrate when compared to other proteins. The VT degradation pattern observed in vitro also showed a similar profile to that observed in vivo. Co-localization of BYC and acidic cortical yolk granules was performed by immunocytochemistry and confocal microscopy. Proton pumping activity of yolk granules in vitro was higher in eggs collected 4 day after oviposition than in newly laid eggs. Taken together, our data suggest that BYC plays a major role in the degradation of VT and that its activity is controlled by acidification of yolk platelets localized at the cortical cytoplasm of the developing Boophilus microplus egg. PMID- 15110866 TI - Functional and comparative analysis of two distinct ecdysteroid-responsive gene expression constructs in Drosophila S2 cells. AB - Inducible expression systems have proven to be of major interest when analysing the function of specific genes or when expressing cytotoxic proteins. In an effort to develop inducible switches allowing for flexible fine-tuning of gene expression levels in insect cells, we have compared the induction capacities of two Drosophila minimal promoters when linked to four consecutive ecdysone response elements. These minimal promoters, either containing a TATA-box or a downstream promoter element, drove the expression of a luciferase reporter gene. Potent induction capacities were observed with the insect moulting hormone, 20 hydroxyecdysone, and with ponasterone A, a plant ecdysteroid. The developed inducible switches further expand the repertoire of molecular tools for functional expression of proteins of interest in insect cells. In addition, the combination of an ecdysone switch with promoters that possess different structural elements can provide novel insights into ecdysteroid-induced transcription in an insect cell line. PMID- 15110867 TI - Regional differences in degree of resilin cross-linking in the desert locust, Schistocerca gregaria. AB - Various cuticular regions from the desert locust, Schistocerca gregaria, were quantitatively analyzed for two cross-linking amino acids, dityrosine and trityrosine, characteristic constituents of the rubberlike cuticular protein, resilin. These amino acids were found in all regions of cuticle investigated, but in widely varying amounts. In fully mature adult locusts the largest amounts of di- and trityrosine were obtained from the prealar arms and wing-hinges, structures possessing long-range elasticity and being involved in energy storage in the flight system. In structures where deformations tend to occur more slowly, such as the clypeo-labral springs and tracheae, di- and trityrosine are less abundant. In sclerotized cuticle from femur and tibia, as well as in cornea and in the highly stretchable intersegmental membranes of mature females, they are only found in trace amounts and are probably unrelated to elasticity. The trityrosine-to-dityrosine ratio in the various cuticular regions vary from nearly equal amounts of the two amino acids to about ten times more dityrosine than trityrosine, indicating that the regions differ in degree of cross-linking; the tracheal wall is the material with the highest trityrosine-to-dityrosine ratio. In some cuticular regions the ratio increases during maturation from newly moulted (teneral) adults to reproductively active locusts; the most pronounced increase was observed for the wing-hinges, and only a small increase was observed for the abdominal tergal plates. In most cuticular regions in fifth instar locust nymphs the contents of di- and trityrosine corresponded to the contents measured for the adult cuticular regions, but only trace amounts of the two amino acids were obtained from the region of the nymphal wing base which corresponds to the wing-hinge containing cuticular region in adult locusts. PMID- 15110870 TI - Identification of an aspartylglucosaminidase-like protein in the venom of the parasitic wasp Asobara tabida (Hymenoptera: Braconidae). AB - This study was designed to identify one of the main components of venomous secretions of the endoparasitic wasp Asobara tabida. By using electrophoretic methods, partial amino acid sequencing and immunostaining, we demonstrated the presence of an aspartylglucosaminidase (AGA)-like protein in the venom of this insect. The enzyme had a polymeric conformation and was formed of 30 and 18 kDa subunits. The relative positions of several amino acids involved in substrate binding and catalytic activity of known AGA-proteins, which are usually lysosomal enzymes, were conserved in the NH(2)-terminal ends of these subunits. Antibodies raised against human AGA recognized the two subunits of the protein and a 44 kDa protein, suggesting the presence of a precursor molecule of the enzyme in the venom. However, no reliable measurement of the AGA activity could be performed on the venom extracts, which could be explained by the fact the enzyme would be stored in the reservoir of the venom apparatus under an inactive form. These results constitute the first description of an AGA-like protein in an insect venom and are discussed with respect to the knowledge acquired on lysosomal and venom enzymes. PMID- 15110869 TI - Negative regulation of prophenoloxidase (proPO) activation by a clip-domain serine proteinase homolog (SPH) from endoparasitoid venom. AB - Most parasitic wasps inject maternal factors into the host hemocoel to suppress the host immune system and ensure successful development of their progeny. Melanization is one of the insect defence mechanisms against intruding pathogens or parasites. We previously isolated from the venom of Cotesia rubecula a 50 kDa protein that blocked melanization in the hemolymph of its host, Pieris rapae [Insect Biochem. Mol. Biol. 33 (2003) 1017]. This protein, designated Vn50, is a serine proteinase homolog (SPH) containing an amino-terminal clip domain. In this work, we demonstrated that recombinant Vn50 bound P. rapae hemolymph components that were recognized by antisera to Tenebrio molitor prophenoloxidase (proPO) and Manduca sexta proPO-activating proteinase (PAP). Vn50 is stable in the host hemolymph-it remained intact for at least 72 h after parasitization. Using M. sexta as a model system, we found that Vn50 efficiently down-regulated proPO activation mediated by M. sexta PAP-1, SPH-1, and SPH-2. Vn50 did not inhibit active phenoloxidase (PO) or PAP-1, but it significantly reduced the proteolysis of proPO. If recombinant Vn50 binds P. rapae proPO and PAP (as suggested by the antibody reactions), it is likely that the molecular interactions among M. sexta proPO, PAP-1, and SPHs were impaired by this venom protein. A similar strategy might be employed by C. rubecula to negatively impact the proPO activation reaction in its natural host. PMID- 15110871 TI - Cloning and expression of a putative transferrin cDNA of the spruce budworm, Choristoneura fumiferana. AB - A spruce budworm (Choristoneura fumiferana) transferrin cDNA (CfTf) was isolated and cloned from a cDNA library that was constructed using mRNA from fifth to sixth instar larvae. CfTf cDNA encoded a predicted protein of 681 amino acids with a molecular mass of approximately 76 kDa. CfTf shared 72% and 74% identities at the amino acid level with transferrins of Manduca sexta and Bombyx mori, respectively. Like other transferrins, CfTf retains most of the N-terminal, iron binding amino acid residues. Northern blot analyses indicated that CfTf mRNA was present at high levels after ecdysis, but that the expression level was low prior to ecdysis at the fourth-sixth instar stages. The highest level of CfTf expression was detected in the fat body. Relatively low levels of expression were detected in the epidermis and no expression was found in the midgut. Expression of CfTf mRNA could be induced by bacteria but not fungi. Expression of CfTf mRNA was suppressed by iron load. PMID- 15110872 TI - Is breast-conserving therapy in the genetically predisposed breast cancer patient a reasonable and appropriate option? PMID- 15110868 TI - The effects of topical application of various fatty acids on pheromone and glandular lipid biosynthesis in the moth Heliothis virescens. AB - Binary mixtures of deuterium-labeled palmitic acid and an excess of different fatty acids were applied to the sex pheromone gland of female Heliothis virescens and the effects on the terminal steps of pheromone biosynthesis, including incorporation of fatty acids into the glandular lipids, observed. Relative to labeled palmitic acid applied alone, application of all the binary mixtures resulted in decreased levels of the labeled pheromone component, (Z)-11 hexadecenyl acetate (Z11-16:OAc), but there was generally no decrease in the amounts of labeled pheromone precursor, (Z)-11-hexadecenoate, nor labeled palmitate in the glandular lipids. These data suggest that the excess of fatty acid in the gland inhibits Delta11-desaturation. However, in the case of excess myristoleic acid, the amount of labeled (Z)-11-hexadecenoate increased significantly, suggesting that this acid inhibited fatty acid reduction. Dose response tests with certain of the fatty acids were consistent with the above interpretations and further indicated that the gland had a high capacity for rapidly activating and incorporating excess fatty acids into the glandular lipids. Finally, application of the various fatty acids resulted in increased levels of these acids in the gland and, in the cases of myristoleic, palmitoleic and myristic acids, it also resulted in increased levels of the corresponding aldehydes, which had previously been detected in the gland of female H. virescens. This suggests that the fatty acid reductase in H. virescens is not highly specific for the major component, and that the final ratio of pheromone components is determined in part by the availability of their corresponding fatty acids in the gland. PMID- 15110873 TI - Best practice in oral care for children and young people being treated for cancer: can we achieve consensus? PMID- 15110874 TI - Breast cancer advocacy across Europe through the work and development of EUROPA DONNA, the European Breast Cancer Coalition. PMID- 15110875 TI - Childhood adrenocortical tumours. AB - Childhood adrenocortical tumours (ACT) constitute only about 0.2% of all paediatric malignancies. However, the incidence of ACT varies across geographic regions and is remarkably high in southern Brazil. At presentation, most children show signs and symptoms of virilisation, which may be accompanied by manifestations of the hypersecretion of other adrenal cortical hormones. Fewer than 10% of patients with ACT show no endocrine syndrome at presentation; these are often older children and adolescents. ACT is commonly associated with constitutional genetic abnormalities, particularly mutations of the P53 gene. Histological features are used to classify the tumours as adenomas or carcinomas; however, the distinction between these two subtypes is often difficult. The extent of disease is best evaluated by computed tomography or magnetic resonance imaging; the role of positron-emission tomographic scans has not been defined. Cure of ACT requires complete tumour resection. The role of chemotherapy or radiotherapy has not been established, although definitive responses to several anticancer drugs have been documented. Among patients who undergo complete tumour resection, favourable prognostic factors include age <4 years, smaller tumour size, signs of virilisation alone at presentation, and adenomatous tumour histology. Some children with ACT show abnormalities of growth and development at the time of presentation, but these usually resolve after surgery. PMID- 15110876 TI - A systematic review of the relation between interleukin-2 schedule and outcome in patients with metastatic renal cell cancer. AB - In Europe, interleukin 2 (IL-2) is one of the two treatment modalities officially approved for patients with metastatic renal cell cancer. Traditionally, IL-2 has been administered by three different routes: intermittent bolus injection (BIV), continuous intravenous infusion (CIV) and subcutaneous injection (SC). There have been few randomized trials designed to compare these routes of administration. This paper describes a systematic review of the literature in which an attempt has been made to determine which schedule of administration is superior. Heterogeneity of the data makes firm conclusions difficult. It appears that the number of complete remissions (CR) is similar between BIV and SC routes and that these are higher than for CIV schedules. The durability of the CRs induced by BIV appeared superior to those induced by SC IL-2 and definitely higher than with CIV protocols. This analysis highlights some of the difficulties of using evidence based medicine to determine standard of care when the clinical-trial data are heterogeneous. These data emphasize the importance of randomized clinical trials in determining what should be regarded as optimum therapy. PMID- 15110877 TI - EUROPA DONNA: has strength in its heterogeneity. AB - EUROPA DONNA, a non-profit-making organisation, is a pan-European movement against breast cancer; it is a coalition that is active in 29 countries, each organised into national independent Fora. In each Forum, a postal survey was conducted between 2001 and 2002. A standardised questionnaire was used to collect information about Forum characteristics and activities, and perceptions of the adequacy of the local health service in relation to the prevention, diagnosis and treatment of breast cancer. Twenty-seven Fora (93%) participated in the survey. The results show the heterogeneity between the Fora in the services offered and activities in the different countries. The Fora also perceived a range of deficiencies in the prevention, diagnosis and treatment of breast cancer within their national health systems. These results highlight the importance of a coalition to establish a consensus among the Fora, and the need for coordinated initiatives in different European countries. PMID- 15110878 TI - Ipsilateral breast tumour recurrence in hereditary breast cancer following breast conserving therapy. AB - The overall rate of an ipsilateral breast tumour recurrence (IBTR) after breast conserving therapy (BCT) ranges from 1% to 2% per year. Risk factors include young age but data on the impact of BRCA1/2 mutations or a definite positive family history for breast cancer are scarce. We investigated IBTR after BCT in patients with hereditary breast cancer (HBC). Through our family cancer clinic we identified 87 HBC patients, including 26 BRCA1/2 carriers, who underwent BCT between 1980 and 1995 (cases). They were compared to 174 patients with sporadic breast cancer (controls) also treated with BCT, matched for age and year of diagnosis. Median follow up was 6.1 years for the cases and 6.0 years for controls. Patient and tumour characteristics were similar in both groups. An IBTR was observed in 19 (21.8%) hereditary and 21 (12.1%) sporadic patients. In the hereditary patients more recurrences occurred elsewhere in the breast (21% versus 9.5%), suggestive of new primaries. Overall, the actuarial IBTR rate was similar at 2 years, but higher in hereditary as compared to sporadic patients at 5 years (14% versus 7%) and at 10 years (30% versus 16%) (P=0.05). Post-relapse and overall survival was not different between hereditary and sporadic cases. Hereditary breast cancer was therefore associated with a higher frequency of early (2-5 years) and late (>5 years) local recurrences following BCT. These data suggest an indication for long-term follow up in HBC and should be taken into account when additional 'risk-reducing' surgery after primary BCT is eventually considered. PMID- 15110879 TI - A cost-utility analysis comparing intensive chemotherapy alone to intensive chemotherapy followed by myeloablative chemotherapy with autologous stem-cell rescue in newly diagnosed patients with stage II/III multiple myeloma; a prospective randomised phase III study. AB - A prospective randomised phase III study in patients < or =65 years old with previously untreated multiple myeloma (MM), intensive chemotherapy followed by myeloablative chemotherapy and autologous stem-cell rescue was compared with intensive chemotherapy alone. This economic evaluation was based on detailed data from patient charts and hospital information systems. In the intention-to-treat analysis, mean total treatment and follow-up costs of the myeloablative treatment arm were 81,643 euros compared to 68,802 euros for the chemotherapy arm (P=0.09). Costs per quality-adjusted life year were 51,357 euros versus 37,328 euros. In the clinical study, no significant differences were found in overall survival after a median follow-up of 33 months from randomisation. Intensive chemotherapy is regarded as standard therapy for younger patients with previously untreated MM. Cost-effectiveness of myeloma therapy after 3 years of follow up seems not to be favoured by myeloablative treatment with autologous stem-cell rescue. PMID- 15110881 TI - S100A9 expression in invasive ductal carcinoma of the breast: S100A9 expression in adenocarcinoma is closely associated with poor tumour differentiation. AB - S100A9 is associated with myelomonocytic cell differentiation and is also expressed in some epithelia. However, there have been few studies on S100A9 in adenocarcinoma (AC) because the expression in normal epithelia is limited to squamous epithelia. Our previous studies on pulmonary AC and liver carcinomas suggested that S100A9 expression in carcinomas of glandular cell origin is related to poor tumour differentiation. In this study, we examined S100A9 expression in invasive breast carcinoma and evaluated the relation of the expression to the tumour differentiation in 70 cases of invasive ductal carcinoma (IDC) of the breast. S100A9 gene and protein expression was detected in MCF-7 breast carcinoma cells. The rate of S100A9 immunopositivity in IDC showed a higher correlation with poor tumour differentiation, especially in nuclear pleomorphism (P=0.0002) and mitotic activity (P=0.0001). Furthermore, transcriptional expression of S100A9 in sections of IDC could be detected in cases with a high S100A9 immunopositivity. No significant differences in the number of myelomonocytic cells expressing S100A9 were found among cases. There was no correlation between S100A9 immunopositivity and lymph node metastasis (P=0.32). S100A9 immunopositivity in non-invasive ductal carcinoma was also associated with poor tumour differentiation. No immunopositive reaction was observed in invasive lobular carcinomas with a classic cytological appearance and non-neoplastic duct cells. We conclude that S100A9 in glandular epithelial cells is newly expressed under cancerous conditions and is over-expressed in poorly differentiated AC. PMID- 15110880 TI - Factors affecting pharmacokinetic variability following doxorubicin and docetaxel based therapy. AB - Current dosing strategies for anticancer drugs result in wide interindividual pharmacokinetic variability. Here, we explored the influence of age, body size, concomitant drugs, dose, infusion duration, and sex on the clearance for doxorubicin and docetaxel in 243 individual patients. Patients received doxorubicin (n=110) or docetaxel (n=152) as monotherapy or in combination chemotherapy regimens. The mean (+/-S.D.) clearance was 63.6+/-22.7 L/h for doxorubicin and 42.8+/-14.9 L/h for docetaxel. Normalisation for body surface area (BSA) reduced the interindividual variability by only <1.7%. Doxorubicin clearance was significantly reduced when administered at doses >50 mg/m(2) or in combination with cyclophosphamide. Upper extremes of body size were associated with increased clearance for both drugs, whereas no consistent effect of age on clearance was discerned. Overall, these findings suggest that incorporation of variables in addition to BSA should be considered in routine dosing strategies for doxorubicin and docetaxel. PMID- 15110883 TI - MAX2--a convenient index to estimate the average per patient risk for chemotherapy toxicity; validation in ECOG trials. AB - Cancer patients, especially the elderly, present with a highly variable susceptibility to toxicity from chemotherapy. To estimate correctly a patient's risk for toxicity, both the average toxicity of a chemotherapy regimen and patient-related variables need to be assessed. However, treatment toxicities are typically reported item by item, not summarised per patient. We tested an index derived from a pilot study, the MAX2, on the ECOG database. Studies including 20 or more patients aged 70 years and older per arm were selected. Four studies were identified, representing 2526 patients, 410 (16%) being elderly. The association of the MAX2 index with the per patient incidence of grade 4 haematological and/or grade 3 or 4 non-haematological toxicity was highly significant, both for the overall group and for the elderly subgroup. The MAX2 index is a convenient and reproducible way of comparing the average per patient risk for toxicity from chemotherapy across several regimens. PMID- 15110884 TI - Factors influencing haematological recovery following high-dose chemotherapy and peripheral stem-cell transplantation for haematological malignancies; 1-year analysis. AB - Peripheral blood counts and factors influencing haematological recovery in 98 patients with a relapse-free survival of > or =1 year treated with high-dose chemotherapy (HDC) and peripheral stem-cell transplantation (PSCT) for haematological malignancies were analysed. One year after PSCT full haematological recovery was demonstrated for haemoglobin (Hb) in 47% of patients, for the white blood cell count (WBC) in 94%, and for platelets in 64%; 39% had a trilineage recovery. In the multivariate analysis, recovery was influenced by age (P=0.002), number of reinfused CD34+ cells (P=0.016), Hb at start of HDC (P=0.001), and platelets at start of HDC (P=0.008). One year following PSCT, 61% of patients still have subnormal values in one or more haematopoietic cell lineage, suggesting a limited bone-marrow reserve. Long-term recovery is highly dependent on age, blood counts at start of HDC and number of reinfused CD34+ cells without a threshold, all reflecting the residual function of bone marrow before HDC. Reinfusing more CD34+ cells can accelerate long-term haematological recovery. PMID- 15110882 TI - Efficacy of droperidol in the prevention of cisplatin-induced delayed emesis: a double-blind, randomised parallel study. AB - We conducted a prospective, randomized, double-blind, parallel study comparing the antiemetic activity and tolerability of treatment with droperidol (2.5 mg d.i.v. twice daily for 5 days) and placebo, both combined with granisetron (3 mg d.i.v. on the first day) and dexamethasone (16 mg d.i.v. on the first day, 8 mg d.i.v. on days 2, 3, and 4 mg d.i.v. on days 4, 5). A total of 180 lung cancer patients receiving high-dose cisplatin (80 mg/m(2))-containing chemotherapy were enrolled in the study, and 171 of them were capable of being evaluated. The clinical characteristics of the patients in the two treatment arms were well balanced. Complete protection from nausea and vomiting was recorded in the acute phase in 97% of patients who treated with droperidol versus 98% of patients who given the placebo (P=0.920), and in 42% versus 38% in the delayed phase (P=0.615). The multiple logistic regression analysis showed that a history of motion sickness was a significant risk factor for cisplatin-induced delayed emesis (odds ratio [OR]=5.98; 95% CI=2.15 to 16.7, P=0.0006). Droperidol containing treatment was well tolerated by most patients, however, the incidence of sleepiness in the droperidol group was higher than in the placebo group (69% versus 30%, P<0.0001). In conclusion, our data did not support the hypothesis that addition of droperidol to granisetron and dexamethasone reduces the delayed emesis induced by high-dose cisplatin. PMID- 15110885 TI - Prevention of oral mucositis in paediatric patients treated with chemotherapy; a randomised crossover trial comparing two protocols of oral care. AB - This study compared the efficacy of two protocols for oral care using either chlorhexidine or benzydamine as oral rinses to alleviate mucositis in children undergoing chemotherapy. Eligible participants were randomised to receive either protocol for 3 weeks in a two-period crossover design. The occurrence of ulcerative lesions and severity of mucositis were measured at baseline and twice weekly, using the modified Oral Assessment Guide (OAG). Data were continuously analysed by plotting them directly on predefined sequential charts. According to this sequential analysis, the study could be terminated at the 34th within subject comparison, with a statistically significant reduction in ulcerative lesions (P<0.05) and severity of mucositis (P<0.05) in children on the chlorhexidine protocol. These findings suggest that chlorhexidine together with oral care might be helpful in alleviating mucositis when given prophylactically to children on chemotherapy, but the therapeutic benefit needs to be confirmed in a larger trial. PMID- 15110886 TI - A survey of current practice with regard to oral care for children being treated for cancer. AB - The aim of the study was to establish current UK oral care practice for children with cancer. A telephone survey of all 22 United Kingdom Children's Cancer Study Group (UKCCSG) centres was undertaken. Nineteen (86%) of the centres reported using guidelines/protocols for mouth care. The use of routine preventive oral care therapies showed the greatest variation between centres. Four centres (18%) did not use any prophylactic oral care therapy other than basic oral hygiene, whereas seven (32%) routinely used a combination of three or more agents. Chlorhexidine was the most frequently administered prophylactic therapy (17/22 centres, 77%), followed by nystatin (11/22 centres, 50%). There was little variation in advice given to parents/patients on basic oral hygiene. Regarding dental check-ups, 9/22 centres (41%) recommended children to attend a hospital linked dental clinic. Only at 8/22 centres (36%) did children undergo a dental check-up before commencing cancer treatment. The survey identified significant variation in preventive oral care therapies and dental check-ups at the UKCCSG centres. Attention needs to be given to establishing evidence based, effective strategies. PMID- 15110888 TI - Can the survival difference between breast cancer patients in Denmark and Sweden 1989 and 1994 be explained by patho-anatomical variables?--a population-based study. AB - Analyses of data from cancer registries have shown a 10% unit difference in 5 year relative survival between Danish and Swedish patients with breast cancer. This study investigates the effect of age and patho-anatomic variables on this survival difference. Hospital records were collected for women over 40 years of age diagnosed in 1989 or 1994 in east Denmark and south Sweden; patho-anatomical variables and survival were compared between 2289 Danish and 1715 Swedish women. Tumours were smaller, node-negative axillae more frequent and well-differentiated tumours almost 10% more frequent in Sweden. A superior 5-year relative survival in Sweden was found in the 50- to 79-year age group. The adjusted hazard rate ratio between countries was 1.7 in 1989 and 1.3 in 1994. Conditional survival after surviving the first 5 years was similar for the two countries. Adjusting for patho-anatomical variables reduced but did not eliminate the higher risk of death among the Danish patients. Higher population death rates could explain some but not all of the residual elevated risk for Danish women. PMID- 15110889 TI - Five-year change in statistical designs of phase II trials published in leading cancer journals. AB - This study compares the evolution in statistical design reporting for phase II cancer clinical trials published in the six following leading journals: American Journal of Clinical Oncology, Annals of Oncology, British Journal of Cancer, Cancer, European Journal of Cancer and Journal of Clinical Oncology. Only articles where tumour response was considered as the primary endpoint were selected. A total of 393 phase II trials published in 1995 (n=185) and 2000 (n=208) were reviewed. Neither sample size nor design parameters were specified in 157 (85%) and 113 (46%) papers in 1995 and 2000, respectively. 28 (15%) and 95 (46%) papers included at least some information on the statistical designs used: Gehan (4.3% and 3.3%), Fleming (2.2% and 4.3%), and Simon (2.7% and 11.0%). Ad hoc, non-referenced methods were used in 5.9% and 27.3% articles in 1995 and 2000, respectively. Although there is an increase in the mention of at least some statistical design parameters in phase II cancer clinical trials over a 5-year period in these selected cancer journals, the use of referenced methods is still short or often inadequate. PMID- 15110887 TI - Detection of cervical precancer and cancer in a hospital population; benefits of testing for human papillomavirus. AB - The aim was to determine the relevance of human papillomavirus (HPV) testing in identifying high-grade cervical intraepithelial neoplasia or worse (CIN2/3+) in a hospital population (n=3574) characterised by a high rate of cytological abnormalities and high-risk HPV infections. According to the results of the initial Papanicolaou and HPV test, women were directly referred for colposcopy/biopsy or recalled for a control visit. Sensitivity and specificity were corrected for verification bias. HPV-testing sensitivity was 94.3%, higher than that of cytological testing at any cut-off point (65.1%-86.8%), while specificity was greater for cytology than for HPV testing (99.3% or 91.8% versus 83.4%). The combination of both tests allowed 100% sensitivity and negative predictive value. We conclude that HPV testing is a relevant tool for the detection of cervical disease. The best way of combining cytology and HPV detection in screening programmes should be evaluated in large-scale studies. PMID- 15110890 TI - Reduced breast cancer risk with increasing serum folate in a case-control study of the C677T genotype of the methylenetetrahydrofolate reductase gene. AB - Breast cancer risk may be associated with folate status or the C677T genotype of the methylenetetrahydrofolate reductase (MTHFR) gene. We compared serum folate concentrations and C677T genotype in 141 breast cancer patients and 109 age matched controls. Serum folate was significantly lower in cases compared to controls (geometric means, 5.7 versus 6.6 microg/l; P=0.005). Breast cancer risk was not associated with C677T genotype. After adjusting for age of menarche, parity, alcohol intake and total fat intake we observed reductions in odds ratios for breast cancer risk comparing the highest with the lowest quartiles of serum folate concentrations of 0.23 (95% confidence interval (CI) 0.09, 0.54) for the entire group, 0.27 (CI 0.09, 0.80) for the wild-type and 0.08 (CI 0.01, 0.52) for the heterozygous C677T genotype. We conclude that for the whole group, and the wild-type and heterozygous C677T genotypes, increased serum concentrations of folate were associated with reduced risks of breast cancer. PMID- 15110891 TI - Molecular analysis of the putative tumour-suppressor gene EXTL1 in neuroblastoma patients and cell lines. AB - Although neuroblastoma is the most common extracranial solid tumour of childhood, little is known about its aetiology. Together with MYCN amplification and chromosome 17q gain, chromosome 1p deletion is one of the most frequently occurring genetic abnormalities in neuroblastoma. Based upon mapping of deletion breakpoints, putative tumour suppressor gene loci have been assigned to the distal part of the short arm of chromosome 1. Recently, the EXTL1 gene was suggested as a candidate neuroblastoma-suppressor gene and to evaluate this hypothesis, we performed 1p deletion analysis and mutation screening of the EXTL1 coding region on DNA from 22 primary neuroblastomas and 21 neuroblastoma cell lines. Deletions of the chromosome region 1p36.1, including the EXTL1 gene, were detected in several neuroblastoma cell lines and primary tumours. EXTL1 mutation screening resulted in the detection of one unclassified variant (Ser28Cys) but could not provide additional evidence of EXTL1 being involved in the aetiology of neuroblastoma. PMID- 15110892 TI - Dynamic contrast-enhanced magnetic resonance imaging at 1.5 Tesla with gadopentetate dimeglumine to assess the angiostatic effects of anginex in mice. AB - The purpose of this study was to evaluate the effects of anginex on tumour angiogenesis assessed by dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) on a clinical 1.5 Tesla MR system and with the clinically available contrast agent gadopentetate dimeglumine. C57BL/6 mice carrying B16F10 melanomas were treated with anginex, TNP-470 or saline. Tumour growth curves and microvessel density (MVD) were recorded to establish the effects of treatment. DCE-MRI was performed on day 16 after tumour inoculation, and the endothelial transfer coefficients of the microvessel permeability surface-area product (K(PS)) were calculated using a two-compartment model. Both anginex and TNP-470 resulted in smaller tumour volumes (P<0.0001) and lower MVD (P <0.05) compared to saline. Treatment with anginex resulted in a 64% reduction (P<0.01) of tumour K(PS) and TNP-470 resulted in a 44% reduction (P=0.17), compared to saline. DCE MRI with a clinically available, small-molecular contrast agent can therefore be used to evaluate the angiostatic effects of anginex and TNP-470 on tumour angiogenesis. PMID- 15110893 TI - Improved penetration of docetaxel into the brain by co-administration of inhibitors of P-glycoprotein. AB - P-glycoprotein (Pgp) in the blood-brain barrier limits the brain's uptake of many anticancer drugs. We have investigated whether the Pgp inhibitors cyclosporin A, valspodar (PSC833) and elacridar (GF120918) increase the accumulation of docetaxel in the brain. Pgp knockout mice served as a reference model for the complete absence or complete inhibition of Pgp. Plasma and tissues were analysed by high-performance liquid chromatography. Cyclosporin A, valspodar and elacridar significantly increased the brain concentrations of docetaxel in wild-type mice to 38%, 56% and 59%, respectively, of those achieved in Pgp knockout mice. Valspodar and cyclosporin A also increased the docetaxel concentration in plasma and other tissues by 2- and 3-fold, whereas elacridar did not change the clearance. All three inhibitors therefore inhibit Pgp in the blood-brain barrier. Elacridar increases the accumulation of docetaxel in the brain without significant effects on systemic exposure. Further clinical tests with this latter combination are warranted. PMID- 15110894 TI - Combination of a CpG-oligodeoxynucleotide and a topoisomerase I inhibitor in the therapy of human tumour xenografts. AB - The study was conducted to investigate the effects of a novel therapeutic approach, i.e. the combination of chemotherapy and immunotherapy, against a human prostate carcinoma xenograft. A topoisomerase I inhibitor, topotecan, and CpG containing oligodeoxynucleotides (CpG-ODN) were combined. Athymic mice bearing the PC-3 human prostate carcinoma were treated with the maximum tolerated dose (MTD) of topotecan (3 weekly treatments) and with repeated treatments of CpG-ODN (40 and 20 microg/mouse); tumour growth and lethal toxicity were monitored. Topotecan effect on CpG-ODN-induced production of interleukin (IL) 12, interferon (IFN)-gamma and tumour necrosis factor-alpha was also assessed. Since topotecan pretreatment differentially influenced CpG-ODN-induced production of IL-12 and IFN-gamma, the antitumour effects of the two therapies were investigated in a sequential (full topotecan regimen followed by CpG-ODN) or in an alternating sequence (starting with CpG-ODN). Topotecan inhibited PC-3 tumour growth, inducing 95% tumour volume inhibition. All combined treatments resulted in a significant delay in tumour growth, compared to the effects in topotecan-treated mice (P<0.01, by analysis of tumour growth curves). The combination regimens were well tolerated, except for the alternating sequence of 40 microg CpG-ODN and topotecan, which resulted in three out of eight toxic deaths. This alternating sequence was highly toxic even when another cytotoxic drug (doxorubicin) was used in healthy mice. In conclusion, the combination of topotecan and CpG-ODN increased antitumour effects over chemotherapy alone in the growth of a human prostate carcinoma xenograft. Administration sequence was critical to the combination toxicity: the complete regimen of the cytotoxic drug followed by repeated administrations of the immunomodulator seemed the most promising for further investigations. PMID- 15110895 TI - Alpha-adducin G460W polymorphism, urinary sodium excretion, and blood pressure in community-based samples. AB - BACKGROUND: There is limited evidence on the gene-environmental interaction among alpha-adducin G460W gene polymorphism, sodium intake, and blood pressure (BP) levels in a general population. One hypothesis is that the association between G460W polymorphism and BP is more evident among persons with higher sodium intake than those with lower sodium intake. METHODS: We conducted a population-based cross-sectional study of 2823 men and women aged 30 to 74 years in a Japanese rural community to examine the association of the alpha-adducin G460W polymorphism with BP levels stratified by salt intake, as estimated by 24-h urine collection and dietary questionnaire. RESULTS: There was no difference in systolic or diastolic BP levels among the GG, GW, and WW groups for women, but for men, mean systolic BP tended to be higher in the WW group than in the GG group. When we stratified men according to sodium excretion/intake, mean systolic BP was significantly higher in the WW group than in the GG group among men with higher urinary sodium excretion (138.8 v 133.6 mm Hg, P =.02) and tended to be higher among men with higher previous sodium intake. No genetic association was found among women or among men with lower urinary sodium excretion or lower sodium intake. CONCLUSIONS: The alpha-adducin WW genotype was associated with higher systolic BP among men with a higher sodium intake. PMID- 15110896 TI - No association of the Asp298 variant of the endothelial nitric oxide synthase gene with preeclampsia. AB - BACKGROUND: Preeclampsia, the most common serious complication of pregnancy, is characterized by vasoconstriction, dysfunction of the vascular endothelium, and hypertension. Unidentified genetic factors and impaired nitric oxide (NO) mediated vasodilation are thought to contribute to the development of the syndrome. Polymorphisms of the endothelial nitric oxide synthase (eNOS) gene affect NO production and have been associated with hypertension and preeclampsia in a Japanese population. METHODS: We compared the frequency of the Glu298Asp eNOS polymorphism in 397 Hispanic and white normotensive pregnant control subjects with the gene frequencies in 64 women with preeclampsia (systolic blood pressure >140 mm Hg or diastolic blood pressure >90 mm Hg, on at least two occasions 6 hours apart, and proteinuria >0.3 g/L or a dipstick proteinuria reading of 2+). RESULTS: Preeclampsia was not associated with the presence of Asp at position 298 of eNOS. CONCLUSIONS: In contrast to the findings in Japanese women, preeclampsia was not associated with the Asp variant of eNOS in an American population. PMID- 15110897 TI - Nitric oxide synthase inhibition accelerates the pressor response to low-dose angiotensin II, exacerbates target organ damage, and induces renin escape. AB - Because nitric oxide may attenuate both the pressor and cytotoxic effects of angiotensin II (Ang II), we investigated whether nitric oxide synthase (NOS) inhibition might accelerate the slow pressor effect of Ang II, and augment target organ damage. Using conscious, chronically catheterized rats, we previously observed that low-dose Ang II (10 ng/kg/min) rapidly increased mean arterial pressure (MAP) by approximately 25 mm Hg. The MAP then remained at this level for 2 to 4 days, and then increased again during the next 5 days by a further 25 mm Hg to a second plateau. In the present study, 7 days of N(omega)-nitro-l-arginine methyl ester (L-NAME; 10 microg/kg/min) alone increased MAP by 16 mm Hg. When Ang II was added to L-NAME, MAP increased as much as with Ang II alone, but then continued to increase until day 4, reaching a plateau as high as that reached only on day 9 of Ang II alone. In approximately half the rats infused with L-NAME + Ang II, plasma renin escaped from Ang II-induced suppression after day 4 of Ang II, and continued to increase for the duration of the study. On the first day that Ang II was added to L-NAME, urinary protein excretion and plasma cardiac troponin T increased, indicating early target organ damage. By the end of the study, all rats treated with L-NAME + Ang II developed tubulointerstitial and glomerular injuries, fibrosis of the renal and cardiac arteries, and cardiac interstitial fibrosis. Target organ damage was greater in rats that developed renin escape than in those in which plasma renin remained suppressed, but was minimal in rats infused with Ang II or L-NAME alone. Taken together, these findings suggest that endogenous NO normally attenuates the pressor response to low-dose Ang II for several days, and protects from Ang II-induced target organ damage. Under conditions of reduced NO bioavailability, which may result from endothelial insufficiency, relatively small changes in circulating Ang II levels may damage target organs. Moreover, renal damage leading to renin escape may initiate a vicious cycle of elevated Ang II production, leading to higher blood pressure and greater target organ damage. PMID- 15110898 TI - Susceptibility of the influence of weight on blood pressure in men versus women: lessons from a large-scale study of young adults. AB - BACKGROUND: Weight and blood pressure (BP) are closely related. The aim of this study was to quantify this relationship and compare it to other factors in a population of relatively young adults, with particular focus on the possible role of gender. METHODS: We conducted a cross-sectional analysis of the Young Adult Periodic Examinations in Israel (YAPEIS) database of healthy people aged 25 to 45 years undergoing routine periodic examinations. Between 1991 and 1999, 38,558 subjects (88.1% men, mean age 36 +/- 8 years) were examined. The correlation between BP and weight was evaluated with adjustments for age, sex, physical activity, cigarette smoking, and fasting blood glucose levels. RESULTS: Blood pressure correlated positively with body mass index (BMI), spanning the spectrum of BMI values. Weight accounted for 8% to 10% of BP variance. The odds ratio for hypertension increased by 16% for each additional unit of BMI, compared to 6% for each year increase in age. The relative propensity of men toward hypertension, typical of this age group, was less pronounced at higher BMI values (male:female ratio = 2.2 at BMI <25 kg/m(2), and 1.28 at BMI > or =35 kg/m(2)). CONCLUSIONS: The association between BP and body weight is at least as strong as that between BP and age and is especially prominent in women. PMID- 15110899 TI - Relationship between changes in neck circumference and changes in blood pressure. AB - OBJECTIVE: Neck circumference (NC), as an upper body obesity index, is a simple screening measure for identifying overweight and obese patients. This study examines a relationship between changes in systolic blood pressure (BP), and diastolic BP and changes in NC, and other components of the metabolic syndrome. METHODS: In a longitudinal cohort study the study group was comprised of 364 subjects (155 men and 209 women) with no known major medical conditions who were not receiving any medication therapy. Main indicators studied included systolic BP, diastolic BP, NC, waist circumference (WC), waist-to-hip ratio (WHR), lipoprotein, glucose, and uric acid levels. RESULTS: Pearson's correlation coefficients indicated a significant association between changes in systolic BP and changes in NC (men, r = 0.54; women, r = 0.56; each, P <.0001), WC men, r = 0.51; women, r = 0.56; each, P <.0001), triglycerides (men, r = 0.41; women, r = 0.44; each, P <.0001), and glucose (men, r = 0.43, P <.001; women, r = 0.48, P <.0001); between changes in diastolic BP and changes in NC (men, r = 0.56; women, r = 0.68; each, P <.0001), WC (men, r = 0.62; women, r = 0.50; each, P <.0001), triglycerides (men, women; each, r = 0.39, P <.0001), and glucose (men, r = 0.45; women, r = 0.49; each, P <.0001). CONCLUSIONS: Changes in systolic BP and diastolic BP correlated positively with changes in NC and other components of the metabolic syndrome. PMID- 15110900 TI - Ramipril in the treatment of hypertension and proteinuria in children with chronic kidney diseases. AB - BACKGROUND: Angiotensin-converting enzyme inhibitors are the drugs of choice in renal hypertension. The efficacy and safety of ramipril in adults has been proved; however, data on effectiveness of ramipril in children are few. The aim of the present study was to investigate the effect of ramipril on blood pressure (BP) and proteinuria in children with chronic kidney diseases. METHODS: A total of 31 children (median age 11.3 years, range 1.9-19.8 years) with various chronic nephropathies and hypertension or proteinuria were prospectively treated with ramipril for 6 months. Blood pressure was evaluated using ambulatory BP monitoring and hypertension was defined as mean BP equal to or greater than the 95th percentile for healthy children. Proteinuria was defined as protein excretion > or =100 mg/m(2)/24 h. The starting dose of ramipril was 1.5 mg/m(2)/24 h once daily. In 27 children it was given as monotherapy. RESULTS: The median decrease in ambulatory BP was 11 mm Hg for daytime systolic, 10 mm Hg for daytime and nighttime diastolic, and 8 mm Hg for nighttime systolic BP. Hypertension normalized in 55% of the children. Proteinuria decreased in 84% of the children with pathologic proteinuria; the median decrease was 51%. A positive correlation was found between initial proteinuria and change of proteinuria (r = 0.95, P <.001). Glomerular filtration rate and serum potassium level did not change significantly. One child developed a cough that was believed to be related to ramipril. CONCLUSIONS: Ramipril is an effective and safe drug in children with chronic kidney diseases associated with hypertension, proteinuria, or both. PMID- 15110901 TI - A longitudinal analysis of antihypertensive drug interactions in a Medicaid population. AB - BACKGROUND: Drug interactions are a frequent cause of adverse drug events. We evaluated whether the frequency of previously reported antihypertensive drug-drug interactions could be reduced by pharmaceutical case management. METHODS: Patients >30 years of age with hypertension who were enrolled in the Iowa Pharmaceutical Case Management (PCM) program were evaluated. All prescription claims for patients were obtained on their date of eligibility and again 9 months later. A drug interaction database was developed to examine potential drug interactions in each patient's regimen. RESULTS: Antihypertensive drugs were taken by 1377 patients at baseline and at 9-month follow-up. Highly significant antihypertensive drug interactions were observed at baseline in 35% of patients (0.47 per patient), and interaction prevalence did not change over time. Decreases in the number of drug interactions tended to occur more commonly among patients of pharmacies that provided the highest intensity of service (11.5% in high-intensity pharmacies v 9% in low- or zero-intensity pharmacies, but this did not achieve statistical significance). Nearly 75% of patients had an interaction of any significance level, and the total number of interactions increased over time (P =.0067). CONCLUSIONS: This Medicaid population with hypertension had a very high prevalence of potential drug interactions. The prevalence of interactions did not change, but the mean number of all interactions actually increased over time. There was some suggestion that higher-intensity pharmacies might be more successful in minimizing the risk of clinically significant drug interactions when compared with lower-intensity pharmacies. PMID- 15110902 TI - Effect of slow-release indapamide and perindopril compared with amlodipine on 24 hour blood pressure and left ventricular mass in hypertensive patients of African ancestry. AB - BACKGROUND: In the treatment of hypertension in subjects of African origins, although hydrochlorothiazide (HCTZ) is not as effective as calcium channel blockers, indapamide is superior to HCTZ. In the present study we therefore compared the effects of slow release (SR) indapamide with the calcium channel blocker amlodipine, when used as initial therapy, on blood pressure (BP) and left ventricular mass (LVM) during 6 months of treatment in this group. METHODS: Patients with a mean daytime ambulatory diastolic BP > or =90 mm Hg and < or =110 mm Hg (n = 125, aged 53 +/- 11 years, 68% women) were randomized to receive open label 1.5 mg of indapamide SR or 5 mg of amlodipine. If daytime ambulatory diastolic BP at 1 month was >/=90 mm Hg, 4 mg of perindopril was added to indapamide SR or the dose of amlodipine was increased to 10 mg. RESULTS: After 1 month of therapy, there was an equivalent decline in systolic and diastolic BP in both groups (P <.0001). In the indapamide-treated group (n = 62) the daytime BP decreased from 153 +/- 12/101 +/- 6 mm Hg to 138 +/- 15/92 +/- 10 mm Hg and for amlodipine (n = 58), it decreased from 152 +/- 13/99 +/- 5 mm Hg to 138 +/- 12/91 +/- 8 mm Hg. At 6 months daytime ambulatory BP decreased to 130 +/- 15/86 +/- 8 mm Hg and to 129 +/- 11/85 +/- 5 mm Hg for the indapamide SR (n = 42) and amlodipine (n = 44) treatment groups, respectively. Both groups showed equivalent regression of LVM index and relative wall thickness. CONCLUSIONS: These data suggest that in hypertensive patients of African ancestry initiating therapy with 1.5 mg of indapamide SR and then adding 4 mg of perindopril is equally as effective as amlodipine therapy at reducing BP, and modifying target organ damage. PMID- 15110903 TI - Myocardial blood flow and efficiency in concentric and eccentric left ventricular hypertrophy. AB - BACKGROUND: It is not clearly understood why concentric left ventricular hypertrophy (increased left ventricular mass and relative wall thickness) is associated with higher cardiovascular risk than eccentric hypertrophy (increased left ventricular mass but normal relative wall thickness). Possible reasons include lower myocardial efficiency or perfusion reserve in concentric than in eccentric hypertrophy. We compared myocardial perfusion reserve and efficiency in normotensive controls and in hypertensive patients with concentric and with eccentric hypertrophy. METHODS: Study subjects comprised 16 patients with hypertension-induced left ventricular hypertrophy and 10 normotensive controls. We measured myocardial perfusion reserve and oxygen consumption by positron emission tomography. We calculated myocardial efficiency by dividing left ventricular minute work by myocardial oxygen consumption. RESULTS: There was no significant difference in myocardial perfusion reserve between patients with concentric (n = 9) as compared to eccentric (n = 7) hypertrophy. However, myocardial perfusion reserve in both patient groups were lower than in controls. Although myocardial efficiency in patients with eccentric hypertrophy and in controls were not different, both values were higher than measurements in patients with concentric hypertrophy (18% +/- 6% v 16% +/- 3% v 13% +/- 4%, eccentric hypertrophy versus controls versus concentric hypertrophy, respectively, P =.04 for both eccentric versus concentric hypertrophy and for controls versus concentric hypertrophy). CONCLUSIONS: Myocardial efficiency but not perfusion reserve is lower in hearts with concentric compared with eccentric left ventricular hypertrophy. This might be an explanation for the higher cardiovascular morbidity and mortality associated with concentric left ventricular hypertrophy. PMID- 15110904 TI - Sleep pulse pressure and awake mean pressure as independent predictors for stroke in older hypertensive patients. AB - BACKGROUND: It remains uncertain which is the stronger predictor for stroke in older hypertensives, ambulatory pulse pressure (PP) or mean blood pressure (MBP). METHODS: We studied the prognosis for stroke in 811 older hypertensives in whom ambulatory BP monitoring was performed. We also assessed silent cerebral infarct (SCI) by brain magnetic resonance imaging. RESULTS: Silent cerebral infarcts were found in 50% of 515 subjects (64% of the total population) in whom we assessed SCI using brain magnetic resonance imaging. During a mean of a 42-month follow-up period, stroke events occurred in 59 subjects. After adjustment for covariates, for each 10 mm Hg increase in sleep PP, there was an independent 43% (95% confidence interval [CI]:16%-75%, P =.001) increase in the stroke risk, and sleep MBP was not a significant factor after controlling for sleep PP. On the other hand, for each 10 mm Hg increase in awake MBP, there was an independent 48% (95% CI: 21%-81%, P =.0002) increase in the stroke risk, and awake PP was not a significant factor after controlling for awake MBP. After adjusting for SCI (a strong predictor, P <.0001) at baseline, the effects of awake MBP (38% risk increase for each 10 mm Hg, P =.007) and sleep PP (32% risk increase for each 10 mm Hg, P =.016) remained significant. CONCLUSIONS: In older hypertensives, the impacts of PP and MBP on stroke risk are different during sleep and awake periods. Sleep PP and awake MBP are both predictors of stroke events independently of SCI. PMID- 15110905 TI - Three novel missense mutations of WNK4, a kinase mutated in inherited hypertension, in Japanese hypertensives: implication of clinical phenotypes. AB - BACKGROUND: Mutations in serine-threonine kinase WNK4 with no lysine (K) at a key catalytic residue cause familial hypertension known as pseudohypoaldosteronism type II (PHAII). The objective of this study was to test whether more subtle changes of WNK4 could be implicated in hypertension or renal failure. METHODS: We screened 956 Japanese patients with hypertension or renal failure for mutations in exons 7 and 17 in the WNK4 gene where the mutations were identified in patients with PHAII. RESULTS: We identified three novel missense mutations, Met546Val (n = 2) and Pro556Thr (n = 2) in exon 7, and Pro1173Thr (n = 1) in exon 17, in a heterozygous state in addition to four single nucleotide polymorphisms including one synonymous mutation (Ala547Ala). Results of genotyping Met546Val and Pro556Thr mutations indicated that these mutations were not present in a Japanese general population (n = 1875). CONCLUSIONS: The present study indicated that a systematic screening of WNK4 in a large set of patients with hypertension or renal failure detected some rare genetic variants. Although substantial contribution of three novel missense mutations in exons 7 and 17 of WNK4 to the genetics of hypertension or renal failure is still unclear, these mutations in the WNK4 gene identified in Japanese hypertensives but not in a general population may contribute to hypertension and progression of hypertensive complications to some extent. PMID- 15110906 TI - Overexpression of Cu/Zn-superoxide dismutase and/or catalase in mice inhibits aorta smooth muscle cell proliferation. AB - BACKGROUND: Increasing evidence demonstrates that reactive oxygen species, for example, superoxide (O(2)(-.)) and hydrogen peroxide (H(2)O(2)), promote vascular smooth muscle cell (VSMC) proliferation, and that superoxide dismutase (SOD) and catalase work in concert to scavenge O(2)(-.) and H(2)O(2). This report examined the effect of overexpressing Cu/Zn-SOD or catalase on epidermal growth factor (EGF)-induced proliferation and mitogen-activated protein kinase (MAPK) phosphorylation in VSMCs. METHODS: The VSMCs were obtained from the aorta of wild type mice and transgenic mice overexpressing Cu/Zn-SOD and catalase in combination or overexpressing Cu/Zn-SOD or catalase alone. The VSMC proliferation was measured by cell counting and bromodeoxyuridine incorporation assay. The MAPK phosphorylation was determined with Western blotting. RESULTS: Treatment of wild type VSMCs with EGF significantly increased proliferation and phosphorylation of extracellular signal-regulated kinases (ERK1/2) and p38 MAPK. Overexpression of Cu/Zn-SOD or catalase attenuated EGF-induced phosphorylation of ERK1/2 and p38 MAPK and suppressed EGF-induced proliferation in VSMCs. For example, the EGF induced phosphorylation of ERK1/2 and p38 MAPK and EGF-induced proliferation in VSMCs overexpressing Cu/Zn-SOD or catalase were significantly less than in wild type VSMCs. Moreover, VSMCs overexpressing Cu/Zn-SOD and catalase in combination showed significantly less proliferation and less phosphorylation of the MAPKs than those overexpressing Cu/Zn-SOD or catalase alone. CONCLUSIONS: Overexpression of Cu/Zn-SOD and catalase in combination is more efficient in inhibiting VSMC proliferation and MAPK phosphorylation than overexpression of Cu/Zn-SOD or catalase alone. PMID- 15110907 TI - Adrenocorticotropic hormone, blood pressure, and serum erythropoietin concentrations in the rat. AB - BACKGROUND: This study was designed to investigate the effects of adrenocorticotropic hormone (ACTH) on systolic blood pressure (BP) and serum erythropoietin (EPO) concentrations in two strains of rats. We hypothesized that ACTH-induced hypertension in the rat is characterized by increased EPO production. METHODS: Male Sprague-Dawley (SD) and Wistar out-bred (Wistar) rats were treated with saline or ACTH (0.2 mg/kg/d). Systolic BP was measured using the tail-cuff method. Serum EPO concentrations were assayed using an ELISA kit for human EPO, which cross-reacts with but underestimates rat EPO. Thymus weight was used as a marker of glucocorticoid activity. RESULTS: In SD rats, ACTH increased systolic BP (from 109 +/- 4 to 142 +/- 5 mm Hg, P <.0005), significantly greater than in saline-treated rats (P <.01). Systolic BP in ACTH treated Wistar increased from 120 +/- 3 to 133 +/- 4 mm Hg (P <.05), but was not significantly different from saline-treated Wistar rats. The ACTH-induced increase in systolic BP was greater in SD than in Wistar rats (P <.05). Serum EPO levels were 5.6 +/- 0.4 in SD and 5.9 +/- 0.3 IU/L in Wistar rats, and decreased to undetectable levels with ACTH treatment in 10 of 10 SD and 7 of 10 Wistar rats. The ACTH treatment increased hemoglobin and hematocrit, and decreased thymus weight in both strains. CONCLUSIONS: 1) ACTH decreased serum EPO concentrations in both strains; 2) EPO is inversely related to ACTH-induced hypertension in the rat; and 3) Wistar rats are relatively resistant to the BP raising effects of ACTH treatment but not to the ACTH-induced decrease in thymus weight. These data suggest that EPO is not causal in ACTH-induced hypertension. PMID- 15110910 TI - Psychological distance to reward: equating the number of stimulus and response segments. AB - Psychological distance to reward, or the segmentation effect, refers to the preference for a terminal link of a concurrent-chains schedule consisting of a simple reinforcement schedule (e.g. fixed interval [FI] 30s) relative to its chained-schedule counterpart (e.g. chained FI 15s FI 15s). This experiment was conducted to examine whether the segmentation effect is due to the number of terminal-link stimulus and response segments per se. Three pigeons pecked under a concurrent-chains schedule in which identical variable-interval (VI) schedules operated in the initial links. In each session, half the terminal-link entries followed one initial-link key and the other half followed the other initial-link key. The initial-link keys correlated with the different terminal links were manipulated across conditions. In the first three conditions, each terminal link contained a chained fixed-time (FT) FT schedule, and in the final three conditions, each terminal link contained a chained FI FI schedule. In each condition, in one terminal link (alternating), the order of two key colors correlated with the different schedule segments alternated across terminal-link entries, whereas in the other terminal link (constant), the order of two other key colors was identical for each entry. With the chained FT FT schedule terminal links, there was indifference between the alternating and constant terminal links within and across pigeons, as indexed by initial-link choice proportions. In addition, terminal-link response rates were relatively low. With the chained FI FI schedule terminal links, for each pigeon, there was relatively more preference for the alternating terminal link and terminal-link response rates increased relative to conditions with the chained FT FT schedule terminal links. These data suggest that the segmentation effect is not due simply to the number of terminal link stimulus or response segments per se, but rather to a required period of responding during a stimulus segment that never is paired with reinforcement. PMID- 15110909 TI - Inter-relationships of microalbuminuria with the other surrogates of the atherosclerotic cardiovascular disease in hypertensive subjects. AB - Clinical and epidemiologic data have demonstrated that microalbuminuria (MA) may legitimately be an integrated marker of cardiovascular risk in nondiabetic hypertensive patients. The relation of augmented urinary albumin excretion rate (UAER) with the other surrogates of the atherosclerotic process, such as carotid artery intima-media thickness, large artery elasticity, and left ventricular hypertrophy gives further support to this view. Available evidence so far indicate that MA possibly reflects a state of increased renal endothelial permeability and may be an easily measured marker of a rather diffuse endothelial dysfunction, low-grade inflammation, and vascular disease burden. Whether management of hypertensive populations may be improved by UAER monitoring and whether the reduction of UAER can decrease cardiovascular morbidity and mortality remains to be determined in the future. PMID- 15110908 TI - Role of putative membrane receptors in the effects of estradiol on human vascular cell growth. AB - The present study was designed to determine whether some of the effects of estrogen on human vascular cell growth are exerted through membrane-binding sites, using native as well as novel protein-bound, membrane non-permeant estrogenic complexes. We measured changes in DNA synthesis and creatine kinase specific activity (CK), after treatment with estradiol-17beta (E(2)), estradiol 17beta-6-(O)-carboxymethyl oxime conjugated to bovine serum albumin (BSA) (E(2) BSA), 6-carboxymethyl genistein (CG) or 6- carboxymethyl genistein bound to the high molecular protein keyhole limpet hemocyanin (CG-KLH), and 7-(O) carboxymethyl daidzein (CD) or 7-(O)-carboxymethyl daidzein linked to keyhole limpet hemocyanin (CD-KLH). High concentrations of either E(2) or E(2)-BSA inhibited DNA synthesis in vascular smooth muscle cells (VSMC) (-39% +/- 28% v 32% +/- 15%). Estradiol as well as CG and CD increased DNA synthesis dose dependently in endothelial ECV-304 cells. The CG and CD, as well as CG-KLH and CD KLH, stimulated DNA synthesis dose dependently in VSMC (66% +/- 2%, 100% +/- 12%, 66% +/- 6%, and 41% +/- 8% at 300 nmol/L, respectively). In contrast all forms of protein-bound hormones were unable to affect DNA synthesis in ECV-304 cells or CK in either cell type. In VSMC, both free and bound hormones increased mitogen activated protein-kinase (MAPK)-kinase activity, which was blocked by UO126, an inhibitor of MAPK-kinase. Furthermore, the effects of E(2), E(2)-BSA, or CG-KLH on DNA synthesis were inhibited by UO126. Using the E(2)-BSA linked to the fluorescent dye Cy3.5, we directly demonstrated the presence of membrane-binding sites for E(2) in VSMC and ECV 304 cells. Hence, the effects of E(2) on DNA synthesis in human VSMC, but not in endothelial cells, are apparently exerted by membrane-binding sites for E(2) and do not require intracellular entry of E(2) through the classic nuclear receptor route. PMID- 15110911 TI - Varying reinforcer duration produces behavioral interactions during multiple schedules. AB - The experiments tested the idea that changes in habituation to the reinforcer contribute to behavioral interactions during multiple schedules. This idea predicts that changing an aspect of the reinforcer should disrupt habituation and produce an interaction. Pigeons and rats responded on multiple variable interval variable interval schedules. Introducing variability into the duration of reinforcers in one component increased response rates in both components when the schedules provided high, but not low, rates of reinforcement. The increases in constant-component response rates grew larger as the session progressed. Within session decreases in responding were smaller when the other component provided variable-, rather than fixed-, duration reinforcers. These results are consistent with the idea that changes in habituation to the reinforcer contribute to behavioral interactions. They help to explain why interactions do not occur for some subjects under conditions that produce them for others. Finally, the results question the assumption that induction and behavioral contrast are always produced by different theoretical mechanisms. PMID- 15110912 TI - Conditioned taste aversion induced by wheel running: further evidence on wheel running duration. AB - Rats given access to a running wheel after drinking a flavored solution subsequently drink less of that liquid. It has been suggested that suppression of intake is the result of conditioned taste aversion (CTA). This study explored whether the magnitude of CTA is related to time in the wheel (i.e., amount of wheel running). During 4 days of conditioning, rats drank an orange liquid for 60 min. Immediately after drinking, experimental rats were transferred to running wheels for either 20 or 60 min. Control animals remained in their home cages. Following the conditioning phase, all rats received a preference test composed of the paired flavored liquid (i.e., orange solution) and water. Rats in both experimental groups (20 and 60 min) decreased their consumption of the orange flavored liquid, but no difference in CTA was found between these groups. Wheel running, whether for 20 or 60 min, suppresses the consumption of a liquid consumed immediately before wheel access. These findings are discussed in terms of discrepancies between CTA induced by wheel running and CTA induced by emetic agents. PMID- 15110913 TI - Presence of the father and parental experience have differentiated effects on pup development in Mongolian gerbils (Meriones unguiculatus). AB - Conflicting data exist on the importance of the father and parental experience during development in rodents. The purpose of this study was to investigate the effects of these two variables on development in Mongolian gerbils. Forty pairs of males and females with a litter size of between 4 and 7 pups were used as subjects. Twenty couples had no experience in raising young. After the birth of their pups, four experimental groups were formed: (I) inexperienced mother and father; (II) inexperienced mother; (III) experienced mother and father and (IV) experienced mother. When the pups reached 10 days of age, pup and parental behavior was recorded in experimental sessions of 15 min on 11 consecutive days. Through the statistical analysis it was found that the presence of the father significantly increased the physical contact between pups and parents and that pups opened their eyes earlier in comparison to the groups without the father. On the other hand, parental experience had a significant influence on the behavior of the pups (locomotion inside and outside the nest, and self-grooming). The results of this study suggest that parental experience and the presence of the father have differentiated effects on development in Mongolian gerbils. PMID- 15110914 TI - Changing tactics: dominance, territoriality, and the responses of "primary" males to competition from conditional breeders in the variegated pupfish (Cyprinodon variegatus). AB - Using variegated pupfish, we examined the flexibility in "primary" male tactics when dealing with the aggressive costs of competition. Analogous to conditional mating strategies, we expected primary males to exhibit one of two interrelated tactics (i.e. dominance or territoriality) in response to different numbers of competitors. In the field, competitors influenced aggression. Primary males defended territories; residents facing more intruders engaged in more chases and obtained fewer spawns per female. In the laboratory, primary males showed dominance at low density, controlling most of the aquarium. With increasing competitor numbers, primary males reduced the area controlled and defended territories. Territories occurred with intermediate to high competitor numbers and only under male-biased sex ratios. During these interactions, aggression was highest and competitors were too many for dominant males to suppress the assertions made by each subordinate to increase its rank. Relinquished control of the entire domain enabled a previously subordinate male to establish a territory in the undefended portion of the aquarium. Reduction in defended area related to a reduction in territorial males' spawning success relative to dominants. These results suggested that primary males, like conditional breeders, would adopt the tactic that enabled them to spawn despite the constraints of competition. PMID- 15110915 TI - State-dependent pairing behaviour in male Gammarus pulex (L.) (Crustacea, Amphipoda): effects of time left to moult and prior pairing status. AB - Because mating can be costly in terms of time and energy, an individual's propensity to engage in courtship and mating activities might be modulated by its physiological state. However, so far, state-dependent mate choice has received little attention The present study examined the effect of both prior pairing status and time left to the moult on the ability of male Gammarus pulex (Crustacea, Amphipoda) to enter in precopula with receptive females. In the lab, males that were freshly collected in precopula pairs in the field had a higher probability of re-pairing and were quicker to enter in precopula with receptive females compared to males of similar size that were freshly collected unpaired. In addition, unpaired males found in the field were closer to their moult than paired males. Considered together, our results strongly suggest that time left to the moult and prior mating status directly influence male propensity to pair in G. pulex. PMID- 15110916 TI - Resistance to change in goldfish. AB - Resistance to change has been studied in several species such as humans, rats, and pigeons. We conducted two experiments using goldfish as subjects to examine the generality of the findings on resistance to change in a phylogenetically more primitive species. In Experiment 1, five goldfish (Carassius auratus) were trained on two-component multiple schedules with different variable-interval schedules in effect. When responding was disrupted by presenting free food during intercomponent intervals or by extinction, resistance to change was greater in the component with the higher reinforcement rates. In Experiment 2, identical variable-interval schedules were presented in two multiple-schedule components, but in one of the components response-independent food was delivered concurrently according to variable-time schedules. Baseline response rates were the same for both components, which is inconsistent with previous findings with other species that the addition of response-independent food decreases response rates. However, response rates in the component with added response-independent food showed the greater resistance to change, which is similar to findings in other species. The convergence of these results across various species confirms the generality of the findings on resistance to change. PMID- 15110917 TI - Safety signals from avoidance learning but not from yoked classical conditioning training pass both summation and retardation tests for inhibition. AB - In one experiment half of the animals were trained to avoid a signaled footshock by jumping (30 or 160 trials), whereas the rest of the animals received the same events as yoked. For all of them the termination of the warning signal and of the shock was followed by a safety signal. Several tests were conducted to assess the ability of the stimuli to suppress licking by measuring the latency in completing 25 consecutive licks in the presence of the stimuli. Fear of the warning signal and inhibitory properties of the safety signal (summation and retardation tests) were measured. The results showed that there were no differences in fear to the warning signal, and that the safety signal behaves as a conditioned inhibitor only for animals trained with a long avoidance procedure, but not in the yoked (classical conditioning) procedure. These results highlight the role played by the avoidance response and its consequences in avoidance learning. PMID- 15110918 TI - Dogs respond appropriately to cues of humans' attentional focus. AB - Dogs' ability to recognise cues of human visual attention was studied in different experiments. Study 1 was designed to test the dogs' responsiveness to their owner's tape-recorded verbal commands (Down!) while the Instructor (who was the owner of the dog) was facing either the dog or a human partner or none of them, or was visually separated from the dog. Results show that dogs were more ready to follow the command if the Instructor attended them during instruction compared to situations when the Instructor faced the human partner or was out of sight of the dog. Importantly, however, dogs showed intermediate performance when the Instructor was orienting into 'empty space' during the re-played verbal commands. This suggests that dogs are able to differentiate the focus of human attention. In Study 2 the same dogs were offered the possibility to beg for food from two unfamiliar humans whose visual attention (i.e. facing the dog or turning away) was systematically varied. The dogs' preference for choosing the attentive person shows that dogs are capable of using visual cues of attention to evaluate the human actors' responsiveness to solicit food-sharing. The dogs' ability to understand the communicatory nature of the situations is discussed in terms of their social cognitive skills and unique evolutionary history. PMID- 15110919 TI - Increased responsiveness of the hypothalamus-pituitary-adrenal (HPA) axis to stress in newborns with atopic disposition. AB - In previous studies, atopic patients showed attenuated cortisol responses to psychosocial stress which is suggestive of a hyporeactive hypothalamus-pituitary adrenal (HPA) axis in this patient group. Regarding the anti-inflammatory role of glucocorticoids, reduced responsiveness of the HPA axis under stress may be one potential explanation of stress-induced exacerbation of atopic symptoms. The present study evaluated whether hyporeactivity of the HPA axis is a feature related to the disposition of atopy rather than a consequence of an ongoing chronic allergic inflammatory process. Newborns with an atopic disposition (parental atopy; n=31) and without atopic disposition (no parental atopy; n=20) were recruited. To further assess atopic disposition, total IgE levels were determined in the cord blood of the neonates. Three days after birth, a blood sample was obtained by a heel prick which is part of a standard pediatric examination. Blood sampling by heel prick is well known to be a significant stressor resulting in activation of the HPA axis in newborns. Analysis of salivary cortisol indicated a significant increase of cortisol levels in the newborns after the stressor with a trend towards an elevated cortisol response in babies with a family history of atopy or with elevated levels of cord IgE (> or = 0.5 kU/l). Neonates with a positive parental atopic heritage and elevated cord IgE were found to show significantly elevated cortisol responses to the heel prick stress when compared to newborns without a parental atopic history and normal cord IgE values. Moreover, cord IgE levels were significantly correlated with basal cortisol levels and the cortisol response to the stressor. These findings suggest that atopic disposition in neonates is associated with altered responsiveness of the HPA axis to stress which may increase the vulnerability to develop manifestation of atopy in later life. PMID- 15110920 TI - Mineralocorticoid and glucocorticoid receptors and their differential effects on memory performance in people with Addison's disease. AB - The effects of corticosteroids on memory performance have been the subject of some controversy. Whilst many studies have shown that high levels of corticosteroids can impair memory performance, others have shown they can facilitate it. One explanation for these discrepant effects arises from the differential activation of the two types of corticosteroid receptor--the mineralocorticoid receptor (MRs) and the glucocorticoid receptor (GRs), and the function each subserves during memory formation. Studies in rats and chickens suggest that activation of the MRs is essential during sensory storage (i.e. encoding), whereas normal levels of activation of the GRs (in addition to the already activated MRs) is essential during memory consolidation and retrieval. By using a repeated measures design with nine people with Addison's disease (mean age, 37.9 years), the effects following activation of the MRs only, GRs only, and a combination of MRs/GRs, on working memory and the episodic and semantic components of declarative memory were investigated. MRs and GRs were activated using either MR specific (9 alpha fluorohydrocortisone) or GR specific (dexamethasone) exogenous steroids, respectively. The results showed that participants performed better in the Digits Backward task when both receptors were activated compared to when GRs only were activated (P<0.01). They also performed better in recall in the Hopkins Verbal Learning Test when both receptors were activated compared to when MRs only were activated (P<0.05) and GRs only were activated (P<0.01). Whilst significant effects were not consistent across memory tasks, these results lend further support to the view that balanced activation of MRs and GRs is necessary for optimal memory function in humans. PMID- 15110921 TI - Levels of DHEA and DHEAS and responses to CRH stimulation and hydrocortisone treatment in chronic fatigue syndrome. AB - BACKGROUND: An association between chronic fatigue syndrome (CFS) and abnormalities of the hypothalamo-pituitary-adrenal axis has been described, and other adrenal steroid abnormalities have been suggested. Dehydroepiandrostenedione (DHEA) and its sulphate (DHEA-S), apart from being a precursor of sex steroids, have other functions associated with memory, depression and sleep. It has been suggested that CFS may be associated with a state of relative DHEA(-S) deficiency. Therefore we investigated basal levels of DHEA(-S), the cortisol/DHEA molar ratio and the responsiveness of DHEA to stimulation by corticotrophin-releasing hormone (CRH). Recent studies have also suggested that low dose hydrocortisone may be effective at reducing fatigue in CFS. We therefore also assessed these parameters prior to and following treatment with low dose oral hydrocortisone. METHODS: Basal levels of serum DHEA, DHEAS and cortisol were measured in 16 patients with CFS without depression and in 16 controls matched for age, gender, weight, body mass index and menstrual history. CRH tests (1 g/kg i.v.) were carried out on all subjects and DHEA measured at 0, +30 and +90 min. In the patient group, CRH tests were repeated on two further occasions following treatment with hydrocortisone (5 or 10 mg, p.o.) or placebo for 1 month each in a double-blind cross over study protocol. RESULTS: Basal levels of DHEA were higher in the patient, compared to the control, group (14.1+/ 2.2 vs. 9.0+/-0.90 ng/ml, P=0.04), while levels of DHEAS in patients (288.7+/ 35.4 microg/dl) were not different from controls (293.7+/-53.8, P=NS). Higher DHEA levels were correlated with higher disability scores. Basal cortisol levels were higher in patients, and consequently the cortisol/DHEA molar ratio did not differ between patients and controls. Levels of DHEA (8.9+/-0.97 ng/ml, P=0.015) and DHEAS (233.4+/-41.6 microg/dl, P=0.03) were lower in patients following treatment with hydrocortisone. There was a rise in DHEA responsiveness to CRH in the patients after treatment but this did not attain significance (AUCc: 2.5+/ 1.7 ng/ml h pre-treatment vs. 6.4+/-1.2 ng/ml h post-hydrocortisone, P=0.053). However, those patients who responded fully to hydrocortisone in terms of reduced fatigue scores did show a significantly increased DHEA responsiveness to CRH (AUCc: -1.4+/-2.5 ng/ml h at baseline, 5.0+/-1.2 ng/ml h after active treatment, P=0.029). CONCLUSIONS: DHEA levels are raised in CFS and correlate with the degree of self-reported disability. Hydrocortisone therapy leads to a reduction in these levels towards normal, and an increased DHEA response to CRH, most marked in those who show a clinical response to this therapy. PMID- 15110922 TI - Heart rate variability in premenstrual dysphoric disorder. AB - Measuring heart rate variability (HRV) is a way to assess the autonomic regulation of the heart. Decreased HRV, indicating reduced parasympathetic tone, has previously been found in depression and anxiety disorders. The objective of this study was to assess HRV in women with premenstrual dysphoric disorder (PMDD). To this end, time domain variables and frequency domain variables were assessed in 28 women with PMDD and in 11 symptom-free controls during both the symptomatic luteal phase and the non-symptomatic follicular phase of the menstrual cycle. Two variables reflecting vagal activity in the time domain, the root mean square of differences of successive normal RR intervals (rMSSD) and standard deviation of normal RR intervals (SDNN) were lower in PMDD patients, but this difference was statistically significant in the follicular phase only. The most important vagal measure in the frequency domain, supine high frequency (HF), also appeared lower in PMDD subjects during the follicular phase. It is suggested that PMDD may be associated with reduced vagal tone compared to controls and that this difference is most apparent in the non-symptomatic follicular phase of the menstrual cycle. PMID- 15110923 TI - Effects of raloxifene and estradiol on hippocampal acetylcholine release and spatial learning in the rat. AB - The effects of raloxifene on acquisition of a delayed matching to position (DMP) T-maze task and on hippocampal acetylcholine release were evaluated and compared with estradiol, to determine whether raloxifene has estrogenic effects on cognitive performance and hippocampal cholinergic activity. Ovariectomized rats received continuous treatment with raloxifene (one of two doses), estradiol, or vehicle for 30 days, followed by behavioral training, and then in vivo microdialysis assessment of basal and potassium-stimulated acetylcholine release. The data show that estradiol significantly enhanced DMP acquisition, whereas raloxifene did not. In contrast, both estradiol and the higher dose of raloxifene significantly increased potassium-stimulated acetylcholine release in the hippocampus. These data suggest that, despite increasing evidence for estrogenic effects of raloxifene in brain, raloxifene does not mimic the effects of estrogen on cognitive performance as assessed by acquisition of a simple spatial memory task in ovariectomized rats. PMID- 15110924 TI - Nicotine inhibition of pulsatile GnRH secretion is mediated by GABAA receptor system in the cultured rat embryonic olfactory placode. AB - In past work, we suggested that nicotine inhibition of in vivo pulsatile LH release is not mediated by opiate receptors known to be involved in the inhibition of LH release. In the present study, we examined whether nicotine inhibits the pulsatile gonadotropin-releasing hormone (GnRH) release, and whether this inhibition of GnRH release by nicotine is mediated by the GABA receptor system, by checking in vitro pulsatile GnRH release from cultured GnRH neurons obtained from olfactory placodes of rat embryos at E13.5. The mean interpulse interval of pulsatile GnRH release into the medium was 34.2+/-2.0 min in the control period and increased to 95.3+/-19.0 min (n=6) in the period of nicotine treatment at a concentration of 500 nM, showing an inhibitory effect of nicotine on pulsatile GnRH release. The GABA(A) receptor antagonist bicuculline used alone at a concentration of 20 microM caused no significant changes in the pulsatile GnRH release, but when used in combination with 500 nM of nicotine, bicuculline blocked the nicotine inhibition of GnRH release. In a separate experiment, nicotine treatment at a concentration of 500 nM significantly increased GABA release. These results suggest that, in the cultured embryonic olfactory placode, nicotine stimulates GABA release, which then inhibits GnRH release through GABA(A) receptor system. PMID- 15110925 TI - Influence of menstrual cycle on platelet serotonin uptake site and serotonin2A receptor binding. AB - Depression and anxiety are common health problems affecting women, particularly during the reproductive years. Major depression is two to three times as common in women than in men. Neuroendocrine factors are likely to contribute to this overall increased risk for developing mood disorders in women, and the neuroendocrine influence is most obviously seen in women with premenstrual dysphoric disorder (PMDD) as these women experience depressed mood and anxiety premenstrually only during ovulatory cycles. Moreover, dysfunction of serotonergic transmission has been regarded as an important mechanism in several psychiatric disorders and ovarian steroids have been shown to profoundly influence the activity of the serotonergic system. Given these facts, the purpose of this study was to examine whether binding of [3H]paroxetine to the platelet serotonin transporter or binding of [3H]lysergic acid diethylamide ([3H]LSD) to the platelet 5-HT2A receptor are influenced by the cyclical changes in circulating estradiol and progesterone that occur during the menstrual cycle. We examined 28 healthy women, without oral contraceptives and with regular menstrual cycles. In the late follicular phase, Bmax for [3H]paroxetine binding was significantly higher than in the ovulatory (p<0.01), early luteal phase (p<0.05) and mid-luteal phase (p<0.01). Bmax for [3H]LSD binding was significantly higher in the early follicular phase and the early luteal phase compared to the mid luteal phase (p<0.001 and p<0.05, respectively). In the early follicular phase and the ovulatory phase, significant correlations between estradiol serum concentrations and Kd for [3H]paroxetine were obtained (p<0.001, respectively). In the luteal phase, significant inverse correlations between progesterone as well as estradiol serum concentrations and Kd for [3H]LSD binding were found (p<0.05, respectively). PMID- 15110926 TI - Altered sensitivity to alcohol in the late luteal phase among patients with premenstrual dysphoric disorder. AB - BACKGROUND: Affective disorders, and possibly also premenstrual dysphoric disorder (PMDD) are risk factors for alcohol abuse in women. Although the majority of prior studies have indicated that alcohol sensitivity does not differ between menstrual cycle phases, patients with PMDD have thus far not been studied. METHODS: We have evaluated the functional sensitivity to a low dose of alcohol in 12 women with and 12 women without PMDD in the mid-follicular and late luteal phases of the menstrual cycle, by comparing the effects of an intravenous alcohol infusion on a number of saccadic eye movement measures, including saccadic eye velocity (SEV), saccade deceleration, and self-rated levels of intoxication. RESULTS: PMDD patients displayed blunted SEV (p<0.01) and saccade deceleration responses (p<0.01) to alcohol infusion in the late luteal phase compared to the mid-follicular phase. Control subjects, on the other hand, did not change their SEV or saccade deceleration responses to alcohol between cycle phases. CONCLUSION: These findings are compatible with altered saccadic eye movement sensitivity in response to alcohol among PMDD patients, particularly in the late luteal phase of the menstrual cycle. PMID- 15110927 TI - Chronic administration of tianeptine balances lipopolysaccharide-induced expression of cytokines in the spleen and hypothalamus of rats. AB - The antidepressant tianeptine has been shown to protect the hippocampus against the deleterious consequences of stress and to attenuate the behavioral and neuroendocrine effects of the cytokine inducer lipopolysaccharide (LPS). Since sickness symptoms are linked to peripheral and brain production of cytokines and pro-inflammatory cytokines can promote neurotoxicity, the present study was undertaken to test the possibility that tianeptine attenuates production of pro inflammatory cytokines. This hypothesis has been tested by studying the effects of a chronic intraperitoneal (i.p.) administration of tianeptine (10 mg/kg twice a day for 21 days) to rats on the induction by LPS (250 microg/kg, i.p.) of the production of pro- and anti-inflammatory cytokines, at the periphery (spleen, pituitary) and in the brain (hypothalamus, hippocampus). The expression of mRNAs coding for IL-1 beta, TNF-alpha, IL-6 or IL-10 (RT-PCR) and plasma levels of IL-1 beta, TNF-alpha and IL-10 (ELISA) were measured at various time intervals following LPS. Chronic tianeptine treatment attenuated LPS-induced expression of TNF-alpha in the spleen as well as plasma levels of this cytokine and altered the central balance between pro- and anti-inflammatory cytokines (IL-1 beta/IL-10). These results open new vistas in the pharmacological activity of tianeptine and provide further insights on the possible mechanisms of action involved in its neuroprotective properties. PMID- 15110928 TI - High serum leptin levels subsequent to weight gain predict renewed weight loss in patients with anorexia nervosa. AB - It has repeatedly been shown that high serum leptin levels at target weight ensue from therapeutically induced weight gain in patients with anorexia nervosa (AN). It was hypothesized that elevated leptin levels may be an important factor underlying the difficulties of maintaining the target-weight in AN patients after re-feeding. The aim of this study was to examine if serum leptin levels at discharge from inpatient treatment predict renewed weight loss within 2 months after discharge and upon a 1 yr follow-up. Univariate variance analysis (ANOVA) revealed that 60% (cor. R2=0.60, P=0.002) of the variance in the BMI standard deviation score (BMI-SDS) 2 months after discharge was explained by the model consisting of the independent variables lg10 leptin levels at discharge (P=0.019) and at admission (P=0.069) and BMI-SDS at admission (P=0.002) and delta BMI between admission and discharge (P=0.047). Similarly, 60% (cor. R2=0.60, P=0.005) of the variance in BMI-SDS 1 yr after discharge was explained by lg10 leptin levels at discharge (P=0.046) and at admission (P=0.052) and BMI-SDS at admission (P=0.008) and 2 months after discharge (P=0.007) and delta BMI between admission and discharge (P=0.933). Patients with a poor outcome after 1 yr (n=9, ANCOVA, group: descriptive P=0.041), but not recovered patients (n=9, P=0.649), had lg10 leptin levels at discharge higher than those of controls when adjusted for BMI and % body fat at discharge. In conclusion, high serum leptin levels at discharge from inpatient treatment may indicate a risk for renewed weight loss and an unfavorable 1 yr outcome in AN. PMID- 15110929 TI - Perceived stress and cortisol levels predict speed of wound healing in healthy male adults. AB - The main purpose of the present study was to investigate the association between perceived stress and impaired cutaneous wound healing in humans using a novel wound assessment technique, and taking into account putative mediating factors such as cortisol levels, health behaviours, and personality factors. The study made use of a prospective, within-subjects design in which 24 male non-smokers participated. Every subject received a standard 4mm-punch biopsy, and the healing progress was monitored via high-resolution ultrasound scanning. Participants completed questionnaires on perceived stress, health behaviours, and personality factors, and sampled saliva for cortisol assessment after awakening at 2 weeks prior, directly after, and 2 weeks after the biopsy. The overall results showed a significant negative correlation between speed of wound healing, and both Perceived Stress scale (PSS) scores (r=-.59; p<.01), and General Health Questionnaire (GHQ) scores (r=-.59; p<.01) at the time of the biopsy. The area under the morning cortisol response curve was negatively correlated with speed of wound healing (r=-.55; p<.05), indicating a clear elevation in the morning cortisol slope of those whose wounds were slowest to heal. A median split of the complete sample yielded that the 'slow healing' group showed higher stress levels (PSS t=3.93, p<.01, GHQ t=2.50, p<.05), lower trait optimism (t=3.25, p<.05), and higher cortisol levels to awakening (F=5.60, p<.05) compared with the 'fast healing' group. None of the health behaviours investigated (i.e. alcohol consumption, exercise, healthy eating, and sleep) were correlated with healing speed at any time point. Our data hint at a considerable influence of stress on wound healing, and suggests that elevated cortisol levels, rather than altered health behaviours, play a role in this effect. PMID- 15110930 TI - Androgenic influences on neural asymmetry: Handedness and language lateralization in individuals with congenital adrenal hyperplasia. AB - This study tested the hypothesis that prenatal androgen levels influence hand preferences and language lateralization, two manifestations of neural asymmetry. Participants were individuals with congenital adrenal hyperplasia (CAH, a genetic disorder that results in excess adrenal androgen production beginning prenatally) (40 females; 29 males) and their unaffected relatives (29 females; 30 males) who ranged in age from 12-45 years. The Edinburgh-Crovitz Inventory and the performance of five simple tasks (the Handedness Activities Test) were the measures of hand preferences, and a dichotic listening task composed of consonant vowel nonsense syllables was the measure of language lateralization. No sex differences were observed among relative controls in hand preferences or language lateralization. Male participants with CAH were less consistently right-handed for writing than unaffected male relatives, when those who had been forced to switch writing hands from left to right were considered with left-handers as being not consistently right-handed. There were no other significant differences between individuals with CAH and unaffected relatives. These results do not support the hypothesis that prenatal androgens influence language lateralization, nor do they support the Geschwind-Behan-Galaburda model that posits a key role for testosterone in the development of cognitive problems in males, secondary to changes in hemispheric development and cognitive lateralization. Hormonal influences on handedness, although not always consistent, may be more likely. However, given that sex differences in both language lateralization and handedness are small, it is possible that limited sample size precludes the detection of consistent group differences. PMID- 15110931 TI - Changes in drug 13C NMR chemical shifts as a tool for monitoring interactions with DNA. AB - The antibiotic drug, netropsin, was complexed with the DNA oligonucleotide duplex [d(GGTATACC)]2 to monitor drug 13C NMR chemical shifts changes. The binding mode of netropsin to the minor groove of DNA is well-known, and served as a good model for evaluating the relative sensitivity of 13C chemical shifts to hydrogen bonding. Large downfield shifts were observed for four resonances of carbons that neighbor sites which are known to form hydrogen bond interactions with the DNA minor groove. Many of the remaining resonances of netropsin exhibit shielding or relatively smaller deshielding changes. Based on the model system presented here, large deshielding NMR shift changes of a ligand upon macromolecule binding can likely be attributed to hydrogen bond formation at nearby sites. PMID- 15110932 TI - Enhancing the staggered fluctuations of an actin filament sliding on Chara myosin. AB - We examined both longitudinal and transversal fluctuations of displacements of an actin filament sliding upon Chara myosin molecules. Although the magnitude of transversal fluctuations remained rather independent of ATP concentration, the longitudinal ones were found to increase their magnitude as the concentration increased. In addition, the longitudinal fluctuations gradually increased as the sliding velocity of the filament increased. PMID- 15110933 TI - Characterization of the photoproducts of protoporphyrin IX bound to human serum albumin and immunoglobulin G. AB - Clinically useful photosensitisers (PSs) are likely bound to subcellular and molecular targets during phototherapy. Binding to a macromolecule has the potential to change the photophysical and photochemical characteristics of the PSs that are crucial for their phototoxicity and cell-killing activity. We investigated the effects of binding of a specific PS (protoporphyrin IX or PPIX) to two proteins, human serum albumin (HSA) and a commercially available immunoglobulin (IgG). These two proteins provide two different environments for PPIX. The albumin binds PPIX in hydrophobic binding sites located in subdomain IIA and IIIA, conversely IgG leaves PPIX exposed to the solvent. We show that photophysical parameters such as emission maxima and fluorescence lifetime depend on the binding site. Our results indicate that the different binding site yields very different rates of formation of photoproducts (more than three times higher for PPIX bound to HSA than to IgG) and that different mechanisms of formation may be occurring. Our characterization shows the relevance of protein binding for the photochemistry and ultimately the phototoxicity of PSs. PMID- 15110934 TI - Influence of local anesthetics on the phosphatidylcholine model membrane: small angle synchrotron X-ray diffraction and neutron scattering study. AB - The phase preferences of egg yolk phosphatidylcholine (EYPC) have been examined in the presence of tertiary amine anesthetics [2-(propyloxy)phenyl]-2-(1 piperidinyl)ethyl ester of carbamic acid (C3A) and [2-(heptyloxy)phenyl]-2-(1 piperidinyl)ethyl ester of carbamic acid (C7A, heptacaine). Using the synchrotron small-angle X-ray diffraction (SAXD), it is shown that the C3A anesthetic induces the cubic and hexagonal (H(I)) phases at 2 > or = C3A:EYPC > 0.5 and H2O:EYPC < or = 40 molar ratios. In contrast, longer alkyloxy chain homolog C7A has no effect on the bilayer arrangement of EYPC at C7A:EYPC < = 1 molar ratios as observed by SAXD in C7A + EYPC mixtures hydrated at H2O:EYPC < = 40 molar ratios, as well as in sonicated C7A + EYPC mixtures hydrated in excess water as proved by the small-angle neutron scattering (SANS). The bilayer thickness d(L) decreases and the bilayer C7A surface area SC7A increases with the increase of C7A:EYPC molar ratio. It is suggested that the ability of tertiary amine local anesthetics to influence the dL and SC7A values and EYPC polymorphism is caused by their effective molecular shape and by charge. The possibility that anesthetic molecules may exert some of their biological effects by virtue of these properties is discussed. PMID- 15110936 TI - Melittin-induced changes in thylakoid membranes: particle electrophoresis and light scattering study. AB - Thylakoids were used as a model system to evaluate the effect of bee venom peptide melittin (Mt) on membrane surface charge. At neutral pH, thylakoid membrane surfaces carry excess negative electrical charge. Mt strongly altered the electrophoretic mobility (EPM) of 'low-salt' thylakoids and did not significantly change the EPM of 'high-salt' thylakoids. Mt increased the primary ionic-exchange processes across the 'low-salt' thylakoid membranes, while it did not affect those of 'high-salt' thylakoids. Mt decreased the proton gradient generation on the membranes at both ionic strengths, but it affected more strongly the 'high-salt' than that of 'low-salt' thylakoids. The primary photochemical activity of photosystem II, estimated by the ratio Fv/Fm, was not influenced by the low Mt concentrations. It decreased only when chloroplasts had been incubated with higher Mt concentrations and this effect was better expressed in 'low-salt' than in 'high-salt' thylakoid membranes. PMID- 15110935 TI - Study in mono and bilayers of the interaction of hepatitis G virus (GBV-C/HGV) synthetic antigen E2(99-118) with cell membrane phospholipids. AB - The interaction of the hepatitis G synthetic peptide E2(99-118) with cell membrane phospholipids of different characteristics such as dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) was studied by Langmuir isotherms. Epifluorescence microscopy and Atomic force microscopy (AFM) was also used to study interactions with DPPC. Compression isotherms of DPPC/E2(99-118) and DPPG/E2(99-118) mixed monolayers showed negative deviation from ideallity consistent with the existence of attractive interactions. The incorporation of the peptide in DPPC monolayer was also confirmed in epifluorescence microscopy and AFM studies. The peptide retarded the formation of DPPC domains and did not let the phospholipid get organized. No important differences in the interactions with DPPC (neutral) or DPPG (anionic) were found, thus suggesting that electrostatics forces do not have a predominant influence in these interactions. PMID- 15110937 TI - Presence of anionic phospholipids rules the membrane localization of phenothiazine type multidrug resistance modulator. AB - Substances able to modulate multidrug resistance (MDR), including antipsychotic phenothiazine derivatives, are mainly cationic amphiphiles. The molecular mechanism of their action can involve interactions with transporter proteins as well as with membrane lipids. The interactions between anionic phospholipids and MDR modulators can be crucial for their action. In present work we study interactions of 2-trifluoromethyl-10-(4-[methanesulfonylamid]buthyl) phenothiazine (FPhMS) with neutral (PC) and anionic lipids (PG and PS). Using microcalorimetry, steady-state and time-resolved fluorescence spectroscopy we show that FPhMS interacts with all lipids studied and drug location in membrane depends on lipid type. The electrostatic attraction between drug and lipid headgroups presumably keeps phenothiazine derivative molecules closer to surface of negatively charged membranes with respect to neutral ones. FPhMS effects on bilayer properties are not proportional to phosphatidylserine content in lipid mixtures. Behavior of equimolar PC:PS mixtures is similar to pure PS bilayers, while 2:1 or 1:2 (mole:mole) PC:PS mixtures resemble pure PC ones. PMID- 15110939 TI - Interaction of 2,2,2-trifluoroethanol with proteins: calorimetric, densimetric and surface tension approach. AB - The thermal denaturation of hen egg-white lysozyme was studied in the presence of 2,2,2-trifluoroethanol (TFE) at various pH values using micro differential scanning calorimetry. Quantitative thermodynamic parameters accompanying the thermal transitions were evaluated. It is observed that thermal unfolding of lysozyme in the presence of TFE upto a concentration of 4.0 mol dm(-3) follows a two-state denaturation mechanism as indicated by the equality of van't Hoff and calorimetric enthalpies. The finer details of interaction were studied by measuring the partial molar volume of some constituent amino acids and glycine peptides from water to aqueous TFE at 298.15 K. The physico-chemical properties of aqueous TFE: apparent molar heat capacities, apparent molar volumes and surface tension were measured to understand the intrinsic properties of the cosolvent as well. From the correlation among the thermal unfolding data on lysozyme in aqueous TFE, calculated preferential interaction parameters, physico chemical properties of aqueous TFE and partial molar volumes of transfer, it is concluded that both solvent mediated effect and direct interaction constitute the mechanism of TFE-protein interactions. PMID- 15110938 TI - Model evaluation for glycolytic oscillations in yeast biotransformations of xenobiotics. AB - Anaerobic glycolysis in yeast perturbed by the reduction of xenobiotic ketones is studied numerically in two models which possess the same topology but different levels of complexity. By comparing both models' predictions for concentrations and fluxes as well as steady or oscillatory temporal behavior we answer the question what phenomena require what kind of minimum model abstraction. While mean concentrations and fluxes are predicted in agreement by both models we observe different domains of oscillatory behavior in parameter space. Generic properties of the glycolytic response to ketones are discussed. PMID- 15110940 TI - The hemoglobin cyanomet ligation analogue and carbon monoxide induce similar allosteric mechanisms. AB - Current thermodynamic models of protein cooperativity predicting sigmoidal ligand equilibrium curves differ in the assumptions regarding the structural/functional properties of the intermediate ligation states. Quantitative information on the intermediates cannot be extracted from the equilibrium curves, but must be obtained from direct studies of the intermediates. Since the intermediates are intrinsically unstable species, ligation analogues with reduced mobility are indispensable tools for cooperativity studies provided that the tertiary/quaternary changes triggered by the ligation analogue are similar to those observed using the physiological ligands. We demonstrate that the valency exchange reactions occurring in mixtures of deoxy and cyanomethemoglobin yield non-random distributions of deoxy/cyanomet intermediates that resemble those observed in the equilibrium with carbon monoxide. Previous and new data using the analogue, in agreement with the studies of the CO intermediates, indicate that the mechanism of hemoglobin cooperativity is neither purely concerted nor sequential nor combinatorial, but contains some elements of each of these models. PMID- 15110942 TI - Isothermal acid-titration calorimetry for evaluating the pH dependence of protein stability. AB - A new method, which can be called as isothermal acid-titration calorimetry (IATC), was proposed for evaluating the enthalpy of protein molecules as a function of pH using isothermal titration calorimetry (ITC). This measurement was used to analyze the acid-denaturation of bovine ribonuclease A. The enthalpy change by acid-denaturation of this protein was estimated as 310 kJ/mol at pH 2.8 and 40 degrees C. This value agreed well with the enthalpy change obtained by differential scanning calorimetry. The midpoint pH and proton binding-number difference observed by IATC agreed well with those of the acid transition of the three-dimensional structure monitored by circular dichroism spectrometry. The van't Hoff enthalpy of the transition was derived from the temperature dependence of the midpoint pH and the proton binding-number difference. It agreed well with the calorimetric enthalpy change directly observed by IATC, strongly indicating that there was no stable intermediate state during the acid transition of this protein. PMID- 15110941 TI - Molecular and functional characterisation of the Serratia marcescens outer membrane protein Omp1. AB - Serratia marcescens outer membrane contains three different general diffusion porins: Omp1, Omp2 and Omp3. Omp1 was cloned and sequenced and it shows a great homology to the family of outer membrane porins that comprises the general porins of enteric bacteria. The gene for Omp1 was transferred into an expression plasmid and was expressed in Escherichia coli UH302 (E. coli UH302 pOM100), a porin deficient strain. Its expression confers a higher susceptibility towards different antibiotics to this strain. Omp1 was purified to homogeneity from outer membrane of E. coli UH302 pOM100. Reconstitution of the purified protein into black lipid bilayers demonstrated that it is a channel-forming component with a single-channel conductance of approximately 2 nS in 1 M KCl similar to that of other porins from enteric bacteria. Omp1 is slightly cation-selective. Its homology to already crystallised members of the family of enteric porins whose three-dimensional-structures are known and allowed the design of a topology model for Omp1. The charge distribution within a porin monomer is similar as in other general diffusion pores. The positively charged amino acids localised at the beta strands opposite the external loop L3, which restrict the pore diameter in the porin monomer. PMID- 15110945 TI - A study on the interaction between 1-decyloxymethyl-3-carbamoylpyridinium salts and model membranes--the effect of counterions. AB - The interaction between 1-decyloxymethyl-3-carbamoylpyridinium salts (PS-X) and two types of vesicles (multilamellar vesicle and sonicated vesicle) was investigated. Vesicles were formed from two classes of phospholipids: 1,2 dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) and 1,2-dipalmitoyl-sn glycero-3-phosphatidylethanolamine (DPPE). The PS-X salts used had nitrate, perchlorate, tetrafluoroborate and halides as counterions. Measurements were carried out using differential scanning calorimetry and 1H NMR. All studied compounds decreased the main phase transition temperatures of both DPPC and DPPE bilayers. All of them also decreased the transition enthalpy of DPPC bilayers, however they had a dual effect on the transition enthalpy of DPPE. Namely, at low concentrations the PS-X salts studied significantly increased the main transition enthalpy of DPPE (perchlorate and tetrafluoroborate the least among them) and decreased it at higher concentrations. We have suggested that surfactant rich and pure domains form on the DPPE bilayer in the presence of PS-ClO4, PS-BF4 and PS NO3, whereas they form on DPPC bilayer only in the presence of PS-ClO4. Results are discussed in terms of counterion molecular geometry and the ability of amide group to form hydrogen bonds with lipids. PMID- 15110943 TI - The non-polar solvent potential of mean force for the dimerization of alanine dipeptide: the role of solute-solvent van der Waals interactions. AB - The non-polar component of the potential of mean force of dimerization of alanine dipeptide has been calculated in explicit solvent by free energy perturbation. We observe that the calculated PMF is inconsistent with a non-polar hydration free energy model based solely on the solute surface area. The non-linear behavior of the solute-solvent van der Waals energy is primarily responsible for the non linear dependence of the potential of mean force with respect to the surface area. The calculated potential of mean force is reproduced by an implicit solvent model based on a solvent continuum model for the solute-solvent van der Waals interaction energy and the surface area for the work of forming the solute cavity. PMID- 15110944 TI - Photophysics of norharmane in micellar environments: a fluorometric study. AB - Steady state photophysics of norharmane (NHM) has been studied in different aqueous micellar environments. In aqueous solution at pH 7, excitation of the neutral species promotes a rapid transfer of proton giving rise to the corresponding cationic emission. Aqueous micelles differing in their surface charge characteristics interact with the fluorophore differently. The dependence of the fluorescence of the probe molecule on different micelles has been exploited to determine the critical micellar concentrations (CMCs) of the surfactants. The binding constant (K) and free energy change (deltaG) for the interaction of norharmane with the micelles have been evaluated from the fluorescence data. The probable location of the probe in the micelles has also been suggested. Polarity of the microenvironment around the probe has been determined for CTAB and TX-100 micellar systems from a comparison of the variation of fluorescence properties of the two prototropic species in water dioxane mixture with varying composition. PMID- 15110947 TI - A quantum mechanical study on phosphotyrosyl peptide binding to the SH2 domain of p56lck tyrosine kinase with insights into the biochemistry of intracellular signal transduction events. AB - A study on the interaction between a phosphotyrosyl peptide with the SH2 domain of Lck kinase has been undertaken with the aid of semiempirical linear-scaling quantum mechanical methods. The structure of this complex has been solved at atomic resolution and, hence, it represents the ideal candidate for studying the charge deformation effects induced by the phosphopeptide on the binding site. Substantial changes in the charge of amino acid residues located in the binding pocket of the protein are observed upon ligand binding. More specifically, our quantum chemical calculations indicate that H-bonds involving charged side-chains are subject to consistent charge deformation effects whereas those forming salt bridges are unaffected by ligand binding. Furthermore, ligand binding has the effect of changing both the magnitude and direction of the protein's macrodipole, which rotates approximately 150 degrees with respect that of the unliganded protein. This suggests that a change in the polarization state of the protein might acts as a switch during the transmission of intracellular signals. The binding energy calculated with the aid of the COSMO solvation model corresponds to about -200 kcal/mol, most of which is attributed to the interaction of the phosphotyrosine head with the amino acid chains located in the binding site of the SH2 domain. PMID- 15110946 TI - Spectroscopic properties, catalytic activities and mechanism studies of [(TpPh)Co(X)(CH3OH)m] . nCH3OH: bicarbonate dehydration in the presence of inhibitors. AB - Two inhibitor-containing 'half-sandwich' cobalt(II) complexes [(TpPh)Co(X)(CH3OH)m] x nCH3OH ((TpPh) = hydrotris (3-phenylpyrazolyl)borate; 1: X- = N3-, m = 1, n = 2; 2: X- = NCS-, m = 0, n = 0) have been synthesized and used as the catalysts in the bicarbonate dehydration reaction. The structures of 1 and 2 were determined by X-ray diffraction analysis, which shows that N3- and NCS- coordinate to the Co(II) ions of 1 and 2, respectively, with the Co-N bond lengths of 1.992(6) A and 1.901(3) A. The coordination geometries of the Co(II) complexes in solution are five-coordinated trigonal bipyramid as revealed by the spectroscopic measurements. The dehydration kinetic measurements of HCO3- are performed by the stopped-flow techniques at pH < 7.9. The apparent dehydration rate constant k(obs) varies linearly with Co(II) complex and H+ concentrations, respectively, and the catalytic activity of 2 is lower than that of 1. The aqua Co(II) complex must be the reactive catalytic species in the catalyzed dehydration reaction and the rate-determining step is the substitution of the labile water molecule by HCO3-. The k(obs) values increase with increasing reaction temperature, and the large negative entropy of activation also indicates the associative activation mode. The inhibition ability of NCS- is stronger than that of N3-, which can be rationalized by the decreases in the Co-N(N3-/NCS-) bond lengths and effective atomic charges of the Co(II) ions based on the X-ray crystallographic data and theoretical calculations in this work. PMID- 15110948 TI - Reduction of nitrogenase Fe protein from Azotobacter vinelandii by dithionite: quantitative and qualitative effects of nucleotides, temperature, pH and reaction buffer. AB - Oxidized Fe protein from Azotobacter vinelandii (Av2(0)) was reduced by dithionite (DT) in the absence and presence of nucleotides, over the temperature range 10-40 degrees C, over the pH range 7-8, and in various buffers--inorganic phosphate, TES, HEPES, and Tris. The reduction of each species of Fe protein- Av2(0), Av2(0)(MgATP)2, and Av2(0)(MgADP)2--was resolved into at least three exponential phases, with relative amplitudes of each phase varying over the range of experimental conditions, suggesting a dynamic population shift of kinetically distinct species. The rapid phase of Av2(0) reduction predominated at low temperature and pH, and in Tris buffer; rapid Av2(0)(MgATP)2 reduction was favored at high temperature and pH, and in phosphate buffer; and Av2(0)(MgADP)2 reduction was favored under more physiologically relevant conditions of 20 degrees C, pH 7.5, and in phosphate buffer. The rates of reduction of Fe protein species did not change with buffer, but temperature and pH do have an effect on the rates. With the appropriate constants, an empirically derived equation estimates the rate of Fe protein reduction at any temperature and pH within the limits 10-40 degrees C and pH 7-8, for a given species of Fe protein, and a given phase of the reaction. At 23.0 degrees C and pH 7.4, the rate of the dominant phase of Av2(0) reduction is 1.9 x 10(8) M(-1) s(-1). Under the same conditions, the rates of the two dominant phases of Av2(0)(MgATP)2 reduction are 1.2 x 10(6) and 1.5 x10 (5) M(-1) s(-1); and the rate of the dominant phase of Av2(0)(MgADP)2 reduction is 3.5 x 10(6) in M(-1) s(-1). Thermodynamic activation parameters for each phase of reduction were calculated. No breaks in the Arrhenius plots for any Fe protein species were observed. PMID- 15110950 TI - [Update on sarcoidosis]. PMID- 15110949 TI - Interpretation of concentration-dependence in aggregation kinetics. AB - Aggregation processes are analyzed by two kinetic models, the random polymerization model and the nucleation-dependent polymerization model. A kinetic equation for the random polymerization model can be derived analytically, revealing the relation between the initial monomer concentration ([M]0), the rate constant (k(a)), time (t), the yield of detectable aggregate ([F]), and the critical aggregation number (m). However, time-course curves for the nucleation dependent polymerization model can be obtained by numerical calculation. It is found that lag time (t(d)) and half-time (t1/2) are proportional to [M](-1) in the random polymerization model, while t(d) and t1/2 are proportional to [M1](-s) (1 < s < n; n is nucleus size) at the lower concentration and are less dependent on [M1] at the higher concentration in the nucleation-dependent polymerization model. PMID- 15110951 TI - [Pulmonary arterial hypertension in systemic sclerosis: definition of a screening algorithm for early detection (the ItinerAIR-Sclerodermie Study)]. AB - PURPOSE: Pulmonary arterial hypertension (PAH) is a severe complication of scleroderma. Its prevalence varies from 5% to 35% in the literature. A systematic yearly screening is recommended for early detection and management of PAH, but no precise algorithm is yet available. METHODS: From literature analysis as well as evaluation of medical needs and practices, a multidisciplinary board of experts proposed an algorithm for the screening of PAH in scleroderma. RESULTS: This algorithm is based on a precise Doppler echocardiography methodology for the purpose of screening scleroderma patients for PAH. Patients are considered as being at high or low risk of PAH depending on the maximal tricuspid regurgitation velocity. High-risk patients undergo right heart catheterization for confirmation of the diagnosis of PAH. A French multicenter transversal observational study ("ItinerAIR Sclerodermie") will be conducted in 21 hospital centers in France and involved 100 investigators organized as multidisciplinary networks. FUTURE PROSPECTS: Final results will provide confirmation that the screening algorithm is applicable in a real world setting, as well as a better knowledge of the prevalence of PAH in the various sub-groups of scleroderma patients, of the risk profile for PAH and of the value of DLCO as a predictive factor for PAH, and will support elaboration of precise screening guidelines. PMID- 15110953 TI - [Diagnosis of cardiac sarcoidosis and follow-up of 24 consecutive patients]. AB - PURPOSE: Cardiac sarcoidosis is responsible for 50% of deaths which mainly occur by ventricular arrhythmia or conduction disorders. The aim of this study is to determine the value of cardiac explorations for an early diagnosis of these localizations, which are often underestimated and can cause sudden death. PATIENTS AND METHODS: We prospectively studied 24 consecutive patients, aged 33 +/-10 years, presenting with a sarcoidosis. Nine (38%) were asymptomatic and had no treatment. Fifteen (62%) were symptomatic: two (8%) had only pulmonary lesions and 13 (54%) had a polyvisceral disease. Seven (30%) were treated. Thirteen (54%) had an elevation of the disease activity markers. The patients had a 12-lead ECG, an echocardiography (TTE), a Holter ECG and a Magnetic Resonance Imaging (MRI) at inclusion. RESULTS: Realization rate was: 100% ECG (24), 83% TTE (20), 75% Holter ECG (18) and 62% MRI (15). Only two patients (8%) had a cardiac involvement. The first one had a polyvisceral sarcoidosis presenting with a hypokinetic cardiomyopathy and a complete AV block and the second one presented with a complete AV block which revealed sarcoidosis. Both patients had a MRI septal hypersignal and disease activity markers. They were treated with cardiac stimulation and corticotherapy: the first patient died suddenly, the second one remains asymptomatic after a 14 months follow-up. The 22 patients (92%) with normal explorations did not present any cardiac involvement during the follow-up (3.7 +/-1.6 years). CONCLUSIONS: This study confirms the rarity of cardiac involvement in sarcoidosis. An exhaustive cardiac check-up does not seem very productive even for patients presenting with polyvisceral disease or an elevation of disease markers. A systematic 12-lead ECG seems to be the most useful and simple tool for the early diagnosis of cardiac sarcoidosis. The other explorations will be realized according to clinical data. The absence of abnormal findings seems to have a good negative predictive value allowing to rule out a cardiac problem. PMID- 15110952 TI - [Splenomegaly in sarcoidosis: clinical features and outcome. Analysis of 17 cases]. AB - PURPOSE: To describe the clinical features, biological datas and outcome of patients with systemic sarcoidosis and splenomegaly. METHODS: A retrospective analysis of 17 patients presenting splenomegaly and sarcoidosis with histological proof. RESULTS: Splenomegaly was clinically perceptible in 13 patients, with a spleen size that extended 4 cm or more below the costal margin in 11 patients. It was painful in five cases. The more frequent clinical features are constitutional symptom (fever in 9 cases) and hepatomegaly (N =7). Chest X-ray showed bilateral hilar lymphadenopathy in nine patients and no abnormality in five cases. Serum angiotensin converting enzyme levels were elevated in 81% of cases. Thrombopenia (N =5) and hypersplenism (N =5) were also observed. Corticosteroid were given to 88% with a good clinical and biological response including a decrease in the spleen volume. Corticotherapy and splenectomy (performed in two patients to rule out lymphoma) didn't change outcome of disease. Sarcoidosis is often chronical (82%) and extensive. CONCLUSION: Splenomegaly may be present in sarcoidosis. Management is not standardized. Corticosteroid is indicated for symptomatic or massive splenomegaly. Splenomegaly is frequently in chronic and extensive sarcoidosis. PMID- 15110956 TI - [Sentinel lymph node detection and clinical applications]. AB - INTRODUCTION: The sentinel lymph node (SLN) procedure consists of finding the first lymph node encountered by lymphatic vessels draining a tumor. This technique identifies the SLN histological status, which is representative of all the other draining area lymph nodes' status. EXEGESIS: SLN identification requires the injection of a lymphatic tracer, which could be either a blue dye or radiolabeled particles, or both. Performing a lymphoscintigraphy, which identifies and quantitates SLNs is a possibility provided by the use of radiolabeled particles before a gamma probe guided search is performed intraoperatively. Finally, a specific histopathological methodology involving serial sections of the entire SLN and immunohistochemistry is also required. CONCLUSION: This technique is applicable to melanoma in which the SLN status might be a stronger prognostic factor than Breslow's thickness; to breast cancer in order to avoid full axillary lymph node dissection in patients with small tumors; and potentially to other carcinomas. PMID- 15110955 TI - [Current treatments of xerophthalmia in Sjogren's syndrome]. AB - PURPOSE: To describe the large variety of treatments currently used in Sjogren's syndrome for one of its major manifestations, keratoconjunctivitis sicca or xerophthalmia. CURRENT KNOWLEDGE AND KEY POINTS: Sjogren's syndrome causes a diffuse immunoinflammatory disturbance of main lacrimal glands and the whole ocular surface. Dry eye syndrome is responsible for chronic and deep impairment of quality of life. Many different tear substitutes have been widely developed that are poorly efficient for relieving patients from their complaints. Tear substitutes of various viscosity from standard artificial tears to synthetic gels may be used. Hyaluronic acid is currently the most promising tear substitute, but all eye drops and gels are only efficient in mild to moderate dry eyes and keratoconjunctivitis sicca mostly resists to lubricants. Moreover, the latter may increase patients' complaints when they are associated to preservatives, antiseptic drugs that have widely demonstrated their toxic or irritating potential. Preservatives are, therefore, to be avoided whenever possible in keratoconjunctivitis sicca, by using monodose disposable packaging or specific bottle filtering or eliminating the preservative. Stimulation of lacrimal and salivary secretions with systemic pilocarpine, or obturation of lacrimal puncta in order to limit the drainage of tears in lachrymal ducts may be useful in most severe forms of Sjogren's syndrome. However, the development of topical cyclosporine and other immunomodulating agents is the most relevant progress in the treatment of keratoconjunctivitis sicca in Sjogren's syndrome. PERSPECTIVES: The future for treating Sjogren's syndrome is most likely to pass through the use of new drugs capable of treating the disease or at least its mechanisms, and not only to try to relieve symptoms with poorly efficient tear substitutes. PMID- 15110954 TI - [Diagnosis of dementia]. AB - PURPOSE: The high prevalence of dementia, particularly of Alzheimer's disease, the increase of their incidence with age, and the population aging make this group of diseases a major problem for public health. Nevertheless, diagnosis is difficult because it depends on evolution of disturbances that patients often cannot precisely relate, on complex neuropsychological explorations, and on pathological examination difficult to obtain. Today in France, geriatricians are fully implicated in the diagnosis of dementia and all the physicians who give care to elderly, should lead easily a diagnosis of dementia. CURRENT KNOWLEDGE AND KEY POINTS: In February 2000 ANAES (French governmental agency for accreditation and evaluation of health system) published recommendations called "practical recommendations for the diagnosis of Alzheimer's disease". These recommendations allow physicians to standardize their practices and consist of a rigorous clinical history and examination, a neuropsychological analysis, standard investigations and application of diagnostic criteria already widely diffused and used. FUTURE PROSPECTS AND PROJECTS: Deepening of knowledge, in particular in the domains of neuropsychology and functional cerebral imagery, should allow physicians to diagnose early dementia. These early diagnosis should allow to initiate a multidisciplinary, preventive and effective care for patients. Specific drugs, that will be available, will be intended mostly for patients with early diagnosis, ideally at a pre-dementia state. PMID- 15110957 TI - [Amiodarone liver toxicity about two cases and review of literature]. AB - INTRODUCTION: Amiodarone can induced hepatic dysfunction. OBSERVATIONS: We describe two patients who developed hepatotoxicity presenting suggestive histological features. The outcome was fatal in one case but lesions were reversible in the second case upon treatment interruption. CONCLUSION: These observations show the necessity of hepatic monitoring of patients treated with amiodarone in order to detect potentially severe hepatotoxicity. PMID- 15110958 TI - [Light chain multiple myeloma revealed by an amyloid arthropathy. A report of two cases]. AB - INTRODUCTION: Amyloid arthropathy occurs in 5-15% of patients with multiple myeloma. It is rarely inaugural. Some cases are reported in which the articular manifestations are present many months before the diagnosis of myeloma. We report two unusual cases where the amyloid arthropathy reveals the multiple myeloma. EXEGESIS: Case 1: A 54-year-old man take medical advice for a polyarthritis evolving for 6 months. He has no evidence of multiple myeloma except a hypercalcemia and a hypoalbuminemia. The diagnosis of multiple myeloma is based on an immunobinding of plasmatic proteins. It is a light chain multiple myeloma, lambda type. Amylosis is revealed by biopsy specimens from the synovial tissue of the knee. The patient died rapidly because of an intestinal hemorrhage. Case 2: A 78-year-old woman complains from a polyarthritis evolving for 4 months. She has besides an elevated erythrocyte sedimentation rate, an anemia and a renal insufficiency, a profound hypogammaglobulinemia. There is no radiological abnormalities. The diagnosis of multiple myeloma is based on serum and urine immunoelectrophoresis. It is a light chain multiple myeloma, lambda type. The patient is at her sixth monthly course of cyclophosphamid and prednisone. As far as amyloid arthropathy is concerned, it is relatively improved by corticoid intra articular injections. CONCLUSION: These are two case reports of a light chain multiple myeloma of lambda type, associated to an amyloid arthropathy in its polyarticular presentation, mimicking a rheumatoid arthritis. The revelatory nature of the amyloid arthropathy is interesting to report. PMID- 15110959 TI - [Dyskeratosis congenita and diffuse lung fibrosis]. PMID- 15110960 TI - [Aortic ulcer: an unusual aortitis]. PMID- 15110961 TI - [Amyloidosis complicating Takayasu's arteritis]. PMID- 15110962 TI - [About the editorial "Geriatric oncology: from conception to clinical practice"]. PMID- 15110964 TI - [Remittent seronegative symmetrical synovitis with pitting oedema revealing systemic lupus erythematous in elderly]. PMID- 15110963 TI - [Minocycline-induced lymphomatoid papulosis]. PMID- 15110967 TI - 1,4,8-trimethylfuro[2,3-H]quinolin-2(1H)-one, a new furocoumarin bioisoster. AB - 1,4,8-Trimethylfuro[2,3-h]quinolin-2(1H)-one (compound 5a) is the most interesting derivative among some new furoquinolinones prepared with the aim of moderating the strong toxic effects of 1,4,6,8-tetramethyl derivative (FQ), a powerful potential drug for photomedicine. Compound 5a showed a photobiological activity lower than FQ, but considerable higher than 8-MOP, the furocoumarin used in clinical photomedicine; contrary to classic furocoumarins, 5a induced a strong inhibition of protein synthesis in mammalian cells. Genotoxicity and skin erythema induction, the main side effects of both FQ and 8-MOP photosensitization, are virtually absent with 5a. This behavior seems to be connected to its particular reaction mechanism: differently from furocoumarin derivatives, 5a induced low levels of DNA-protein and no inter-strands cross links, but formed covalent RNA-protein linkages, lesions not observed with known furocoumarins. Moreover, compound 5a generated reactive oxygen species to a considerable extent. For these features, compound 5a appears to be a new photosensitizing agent whose special activity deserves to be deeply investigated. PMID- 15110965 TI - Synthesis, immunomodulating activity and (1)H NMR studies of 7-oxo-9,11-ethano-13 azaprostanoids. AB - Novel 9,11-ethano analogues of prostaglandin endoperoxides with a nitrogen in position 13 were synthesized. (1)H NMR spectra of the obtained compounds were studied. All prostanoids administered perorally at doses of 2.5-10.0 microg x kg( 1) had specific dose-dependent effects on the B-cellular immunity estimated under in vivo conditions on the model of the B-cellular immune response. In terms of the direction of their activities, eight of the studied compounds were found to be immunostimulators, whereas other three compounds displayed immunosuppressing effect. Two of the compounds increased the amount of antibody-forming cells (AFC) per 10(6) spleen cells by 1.9 times in comparison with the respective parameter of control group. PMID- 15110968 TI - Design, synthesis and biological evaluation of new potent 5-nitrofuryl derivatives as anti-Trypanosoma cruzi agents. Studies of trypanothione binding site of trypanothione reductase as target for rational design. AB - Design, using force-field calculations on the catalytic site of trypanothione reductase from Trypanosoma cruzi, has led to the development of new 5-nitrofuryl derivatives as potential anti-trypanosomal agents. The synthesized compounds were tested in vitro against T. cruzi and more than 75% of the prepared derivatives showed higher activity than nifurtimox. Compounds 5 and 11, hexyl 4-(5 nitrofurfurylidene)carbazate and N-hexyl 3-(5-nitrofuryl)propenamide, showed the highest in vitro trypanocidal effect reported to date for members of the nitrofuran family. Partition coefficients and energies for the single-electron reduction of compounds were theoretically determined. These properties could be not the major cause of the activities' differences. The physicochemical environment around E19, W22, C53 and Y111 residues within the trypanothione binding site of trypanothione reductase resulted a valuable target for the rational design of anti-trypanosomal drugs. PMID- 15110970 TI - Synthesis and antiinflammatory activity of heterocyclic indole derivatives. AB - Chalcones of indole 1-5 and their corresponding products; pyrazolines 6-10 and azo compounds 11-15 were synthesised and evaluated for their antiinflammatory activity against carrageenan induced oedema in albino rats at a dose of 50 mg x kg(-1) oral. The structure of compounds was confirmed by IR, (1)H-NMR and mass spectral data. All the compounds of this series showed promising antiinflammatory activity. The most active compound of this series is 3-[1-acetyl-5-(p hydroxyphenyl)-2-pyrazolin-3-yl]indole (7) was found to be most potent, which has shown higher percent of inhibition of oedema, lower ulcerogenic liability and acute toxicity than the standard drug phenylbutazone. PMID- 15110966 TI - Synthesis and in vitro inhibitory activity on human platelet aggregation of novel properly substituted 4-(1-piperazinyl)coumarins. AB - Pursuing our chemical and biological studies in this field, we described the multistep preparation of the new 5-, 6-, or 7-alkoxy and 7-alkoxy-8-methyl substituted 4-(1-piperazinyl)coumarins 5d-v, as well as the in vitro evaluation of their inhibitory activity on human platelet aggregation induced in platelet rich plasma by ADP, collagen or the Ca(2+) ionophore A23187. Compounds 5h-j,p,r-u showed notably high activity towards all the platelet aggregation inducers used, and the most active one, 8-methyl-4-(1-piperazinyl)-7-(3-pyridylmethoxy)coumarin (5t), proved to be a potent in vitro antiplatelet agent. PMID- 15110969 TI - Synthesis and biological activity of novel substituted pyridines and purines containing 2,4-thiazolidinedione. AB - A series of substituted pyridines and purines containing 2,4-thiazolidinedione were designed and synthesized from their corresponding pyridines and purines. These synthesized compounds (entry no. 6a-d, 12a-e, 18a-d, 23a-c) were evaluated for their effect on triglyceride accumulation in 3T3-L1 cells in vitro and their hypoglycemic and hypolipidemic activity in the genetically diabetic KKA(y) mice in vivo. On the basis of their biological activities, 5-(4-[2-[N-methyl-(5-phenyl pyridin-2-yl)amino]ethoxy]benzyl)thiazolidine-2,4-dione (6d) was selected as a candidate for further pharmacological studies. PMID- 15110971 TI - Synthesis and biological evaluation of oxindoles and benzimidazolinones derivatives. AB - The synthesis of new oxindoles and benzimidazolinones derivatives bearing a sugar residue on the aromatic nitrogen is described. The presence of the glycoside moiety should enhance the solubility of these heterocyclic compounds and/or improve the interaction with the active site of the biological targets. The inhibitory activities of these new compounds toward five kinases were examined: KDR (VEGFR-2), FGFR-1, PDGFR-beta, EGFR and Tie 2. Furthermore, the antibacterial activities of the prepared compounds were tested against two Gram-positive bacteria Bacillus cereus and Streptomyces chartreusis, a Gram-negative bacterium Escherichia coli and a yeast Candida albicans. PMID- 15110974 TI - Matrix metalloproteinase inhibition in corneal ulceration. AB - The primary objective of current treatment strategies for infectious keratitis is to sterilize the ulcer as rapidly as possible with topically administered antibiotics. Ulcerative processes can proceed in some cases, despite the absence of microbes, as a result of remaining corneal and tear film MMPs. Combining antibiotic therapy with MMP inhibitors can speed corneal healing, because MMPs play an important role in corneal ulceration and stromal liquefaction. MMPs from the rabbit, horse, and human being are inhibited by metal-binding agents EDTA, NAC, and doxycycline as well as by the serum antiprotease alpha2-macroglobulin. It is not yet certain which proteinase inhibitor has the most favorable therapeutic index for clinical use, although we prefer serum because of its effects on multiple types of proteinases. The MMP inhibitors do have significant therapeutic promise in the treatment of corneal ulceration. PMID- 15110975 TI - Topical therapeutic agents that modulate corneal wound healing. AB - In summary, corneal wound healing is a complex phenomenon that involves interplay between the cellular elements of the cornea, numerous soluble factors, and the constituents of the ECM. Unfortunately, many studies that demonstrate marked alteration on cell behavior in vitro and even in in vivo experiments are often not helpful in the diseased patient, as shown by the results of careful clinical trials. Future work that addresses the complex milieu of the corneal wound healing environment by addressing the interaction of many of these factors will be more likely to be successful than seeking a single agent that will enhance wound healing in all situations. Modulation of wound healing processes by the application of topical therapeutic agents is, however, an expanding field of study sure to produce clinically significant improvements in the management of veterinary patients with corneal defects. The judicious use of topical cytoactive compounds has a place in the clinician's armamentarium integrated into a therapeutic plan that decreases the mechanical stresses imposed on the wound bed as well as removal of any underlying inciting cause. PMID- 15110976 TI - Antiviral therapy for ocular viral disease. AB - Ophthalmic manifestations of viral disease are commonly encountered in veterinary practice. Although the number of antiviral agents is rapidly increasing, the efficacy of many of these drugs against animal viral pathogens may not be known. Furthermore, some of these newer medications may, in fact. be toxic to animal patients. Continued research on the efficacy of these medications in treating viral diseases of veterinary importance is warranted.A thorough review of the ever-growing body of literature is imperative before instituting any new or unknown antiviral therapy. At this time, feline HSV-1 infections remain the most well understood of the ocular viral infections that veterinary practitioners may be called on to treat. Ironically, the number of antiviral agents proven to be effective in naturally occurring clinical cases of FHV-I remains quite limited. With new medications being investigated, this paucity of information on efficacious antiviral therapeutic agents will hopefully improve. PMID- 15110973 TI - Influence of trans-1,2-diaminocyclohexane structure and mixed carboxylic/phosphonic group combinations on samarium-153 chelation capacity and stability. AB - A simple procedure was developed to compare chelating agents for (153)Sm complexes as a preliminary step to synthesise bifunctional analogues. Several variables affecting the efficiency of complex stability were investigated, such as the pre-organisation concept, cavity size, and the nature of coordination sites. Four semi-rigid agents incorporating carboxylic and/or phosphonic groupings fixed at trans-1,2-diaminocyclohexane were evaluated for their (153)Sm chelation properties, and competition studies were performed. Data on the stability of the best chelating agent compound 3: trans-cyclohexane-1,2 bis(aminomethylphosphonic)-N,N'-bis(ethyl-2-iminodiacetic acid) in human serum are presented. PMID- 15110972 TI - synthesis, characterization and in vitro antiamoebic activity of 5-nitrothiophene 2-carboxaldehyde thiosemicarbazones and their Palladium (II) and Ruthenium (II) complexes. AB - Synthesis of new Palladium(II) and Ruthenium(II) complexes of the type, [Pd(L)Cl(2)] and [Ru(eta(4)-C(8)H(12))(L)Cl(2)] [where, L = thiosemicarbazones derived from 5-nitrothiophene-2-carboxaldehyde and cycloalkylaminothiocarbonyl hydrazines] have been isolated by the reaction of [Pd(DMSO)(2)Cl(2)] and [Ru(eta(4)-C(8)H(12))(CH(3)CN)(2)Cl(2)] with 5-nitrothiophene-2-carboxaldehyde thiosemicarbazones. The spectral data revealed that the thiosemicarbazones act as bidentate ligands, making use of thionic sulphur and the azomethine nitrogen atom for coordination to the central metal ion. Microdilution method was used for the assessment of antiamoebic activity of all the compounds against HK-9 strain of Entamoeba histolytica. Among all the thiosemicarbazones, 5-NT-4-BPTSCN (3) showed significant antiamoebic activity (IC(50) - 2.56 microM). Enhancement of antiamoebic activity resulted by introducing palladium and ruthenium metals in the thiosemicarbazone moiety. All the Pd(II) and Ru(II) complexes of 5 nitrothiophene-2-carboxaldehyde thiosemicarbazones were found more active then their respective ligands. The complexes 1a-4a, 1b and 3b showed antiamoebic activity. PMID- 15110977 TI - Antibacterial agents for ocular therapeutics. AB - Ocular bacterial infections of animals take a variety of forms. The eyelid, external ocular surface, anterior segment, posterior segment,and orbit are all frequent targets. Effective management of ocular bacterial infections requires specific identification of the offending organism(s) combined with knowledge of the mechanisms of action and the therapeutic characteristics of available antibiotics. PMID- 15110978 TI - Antifungals and their use in veterinary ophthalmology. AB - Many variables affect the outcome of keratomycosis and systemic fungal infections in animals. These include pathogenicity of the fungal organism(toxins, trophisms, and evasion of host response); previous treatment with topical or systemic corticosteroids, which can have a dramatic negative impact on host defense mechanisms: concurrent systemic illness or immunocompromise: severity/extent of infection; and degree of pain (ie,increased reflex tearing dilutes topical medication) [14]. Experimental work suggests that antibiotics may occasionally exacerbate fungal infections [142],and some researchers advocate that concurrent antibiotic therapy is contraindicated in horses with yeast infections and septate fungal infections unless bacterial infection is also suspected [14]. Nevertheless, given that normal conjunctival flora often include bacteria and fungi and because care of keratomycoses often includes mixed bacterial and fungal infections, the possible dynamics (natural influences and local competition) between ocular surface microorganisms merit further investigation. There are many unanswered questions regarding the accuracy of in vitro susceptibilities and corneal concentration capabilities for antifungal topical medications [14]. Inherent host resistance or other immune interactions between the patient and fungus are perhaps the most important determinants of the outcome but are currently difficult to measure or assess except by subjective clinical observation [14]. PMID- 15110979 TI - The use of corticosteroids to treat ocular inflammation. AB - Corticosteroids are invaluable therapeutic agents for treatment of ocular inflammation in small animal patients. The use of potent anti-inflammatory agents carries with it the risk of some side effects, however. Although some of these may be lessened by topical or subconjunctival administration, these routes are associated with specific ocular side effects about which the practitioner must be aware. With judicious use, corticosteroids remain a mainstay for the prevention and treatment of many painful and potentially blinding ocular diseases. PMID- 15110980 TI - Nonsteroidal anti-inflammatory drugs in veterinary ophthalmology. AB - Uveitis is a common sequela to many ocular diseases. Primary treatment goals for uveitis should be to halt inflammation, prevent or control complications caused by inflammation, relieve pain, and preserve vision. Systemic and topical NSAIDs are essential components of the pharmaceutic armamentarium currently employed in the management of ocular inflammation by general practitioners and veterinary ophthalmologists worldwide. NSAIDs effectively prevent intraoperative miosis; control postoperative pain and inflammation after intraocular procedures, thus optimizing surgical outcome; control symptoms of allergic conjunctivitis;alleviate pain from various causes of uveitis; and circumvent some of the unwanted side effects that occur with corticosteroid treatment. Systemic NSAID therapy is necessary to treat posterior uveitis, because therapeutic concentrations cannot be attained in the retina and choroid with topical administration alone, and is warranted when diseases, such as diabetes mellitus or systemic infection, preclude the use of systemic corticosteroids. Risk factors have been identified with systemic and topical administration of NSAIDs. In general, ophthalmic NSAIDs may be used safely with other ophthalmic pharmaceutics; however, concurrent use of drugs known to affect the corneal epithelium adversely, such as gentamicin, may lead to increased corneal penetration of the NSAID. The concurrent use of NSAIDs with topical corticosteroids in the face of significant preexisting corneal inflammation has been identified as a risk factor in precipitating corneal erosions and melts in people and should be undertaken with caution[8]. Clinicians should remain vigilant in their screening of ophthalmic and systemic complications secondary to drug therapy and educate owners accordingly. If a sudden increase in patient ocular pain (as manifested by an increase in blepharospasm, photophobia, ocular discharge, or rubbing)is noted, owners should be instructed to contact their veterinarian promptly. PMID- 15110981 TI - Immunomodulating agents. AB - Inflammation involves a universally recognized, although incompletely understood, cascade of molecular events orchestrated by lymphokines and other innate biochemicals of immunity. Repeated or extended contact with immunogenic agents results in adaptive immunity involving antigen-induced events that stimulate down stream immune cells and result in expansion of the inflammatory cascade. When immunogenic stimulation persists or autoregulatory immune mechanisms go awry, however, adaptive immunologic events can result in immune-mediated processes detrimental to systemic or organ-specific homeostasis. Because of the complexities of immunologic events, the potential side effects of long-term corticosteroid therapy, and the focused spectrum of most conventional nonsteroidal anti-inflammatory agents (centered on arachidonic acid-related mechanisms), a variety of other chemotherapeutic immunosuppressive agents have assumed an increasingly prominent therapeutic role in veterinary ophthalmology in the management of chronic ocular inflammatory diseases. In addition, nonimmunosuppressive immunomodulating agents (ie, immuno-stimulants or immunorestoratives) may be used as adjunctive therapies in the management of ocular or visual system diseases. PMID- 15110983 TI - Ocular hypotensive drugs. AB - The glaucomas are vision-threatening diseases that commonly result in blindness. In addition to knowledge of the at-risk breeds and predisposing factors for glaucoma as well as the clinical signs of the disease, regular IOP screening with tonometry is required to confirm glaucoma and to monitor the status of the eye. Early intervention with a combination of medications that are capable, in a given species, of reducing aqueous production and increasing aqueous outflow as well as potentially reducing progressive neurosensory degeneration is key to the medical management of this disease. PMID- 15110982 TI - Lacrimostimulants and lacrimomimetics. AB - A thorough understanding of tear film physiology and the clinical manifestations of tear film abnormalities enables the veterinarian to diagnose and treat quantitative (decreased aqueous layer) and qualitative (decreased mucin or lipid layers) tear film abnormalities accurately and to monitor the responses to lacrimostimulatory and lacrimomimetic therapy. This article reviews the embryology,anatomy, and physiology of the lacrimal glands; glands of the nictitating membrane; goblet cells; and tarsal glands as well as the pathophysiology of tear film deficiencies. We also review lacrimo-stimulants, including cyclosporine, tacrolimus, sirolimus, pilocar-pine, and lacrimomimetics (tear film replacements). PMID- 15110985 TI - Agents for intraocular surgery. AB - No single viscoelastic material has all the properties required for every surgical procedure. Ophthalmic surgeons should be familiar with the advantages and disadvantages of several viscoelastics and realize the limitations encountered if the surgeon chooses to rely on a single viscoelastic. It is possible in the future that newer viscoelastics may become available offering aspects of cohesive and dispersive properties that make them easy to remove and not associated with postoperative hypertension. Investigation into other types of viscoelastic materials, such as poly-acrylamide (Orcolon) [33,51,52], human collagen [33,53], polytriethylene-glycol monomethacrylate, and polyglycerol monomethacrylate [48], may result in the availability of synthetic viscoelastic materials. Rethinking how we use viscoelastics may also be indicated. The approach of a single or repeat instillation of a viscoelastic only to have it aspirated during the procedure could be modified by using a continuous infusion ofa dilute viscoelastic [54]. One study diluted hyaluronic acid to a concentration of 0.06% to 0.12% and maintained continuous infusion of the viscofluid through the PE irrigation line [54]. It has been suggested that this may reduce turbulence and improve tissue protection [54]. PMID- 15110984 TI - Nonhypotensive autonomic agents in veterinary ophthalmology. AB - The parasympathetic and sympathetic divisions of the autonomic nervous system are involved in homeostatic control of a wide variety of ocular functions, including accommodation, pupillomotor control, lacrimation, eyelid position, and aqueous humor production. Familiarity with the functional anatomy of the autonomic nervous system is paramount to the understanding and application of the large number of autonomic drugs used in veterinary ophthalmology. The cholinergic and adrenergic agents discussed in this article are commonly employed to facilitate routine ophthalmic examination, in the diagnosis of autonomic dysfunction, and in the treatment of a variety of ocular diseases. PMID- 15110986 TI - Compounding in veterinary ophthalmology. AB - Compounded medications provide treatment options that can improve the clinical outcome for patients, improve compliance, and increase the potential for cure. This article provides an overview of the types and uses of compounded ophthalmic products in veterinary practice as well as criteria for selecting a pharmacist skilled in sterile compounding. PMID- 15110987 TI - Ocular therapeutics. PMID- 15110988 TI - Shuttling components of nuclear import machinery involved in nuclear translocation of steroid receptors exit nucleus via exportin-1/CRM-1* independent pathway. AB - The nucleocytoplasmic transport processes are mediated by soluble transport factors constantly navigating between nuclear and cytoplasmic compartments. Our understanding about nuclear export of general 'nuclear import factors' that deliver the cargo to the nucleus is still fragmentary. Utilizing green fluorescent protein tagged glucocorticoid receptor (GR) and relA as our working model and with judicious use of LMB, we show in living cells that all the soluble components of the nuclear import machinery exit nucleus via exportin1/CRM1 independent pathway(s). PMID- 15110989 TI - Effects of Li+ transport and intracellular binding on Li+/Mg2+ competition in bovine chromaffin cells. AB - Li(+) transport, intracellular immobilisation and Li(+)/Mg(2+) competition were studied in Li(+)-loaded bovine chromaffin cells. Li(+) influx rate constants, k(i), obtained by atomic absorption (AA) spectrophotometry, in control (without and with ouabain) and depolarising (without and with nitrendipine) conditions, showed that L-type voltage-sensitive Ca(2+) channels have an important role in Li(+) uptake under depolarising conditions. The Li(+) influx apparent rate constant, k(iapp), determined under control conditions by (7)Li NMR spectroscopy with the cells immobilised and perfused, was much lower than the AA-determined value for the cells in suspension. Loading of cell suspensions with 15 mmol l(-1) LiCl led, within 90 min, to a AA-measured total intracellular Li(+) concentration, [Li(+)](iT)=11.39+/-0.56 mmol (l cells)(-1), very close to the steady state value. The intracellular Li(+) T(1)/T(2) ratio of (7)Li NMR relaxation times of the Li(+)-loaded cells reflected a high degree of Li(+) immobilisation in bovine chromaffin cells, similar to neuroblastoma, but larger than for lymphoblastoma and erythrocyte cells. A 52% increase in the intracellular free Mg(2+) concentration, Delta[Mg(2+)](f)=0.27+/-0.05 mmol (l cells)(-1) was measured for chromaffin cells loaded with the Mg(2+)-specific fluorescent probe furaptra, after 90-min loading with 15 mmol l(-1) LiCl, using fluorescence spectroscopy, indicating significant displacement of Mg(2+) by Li(+) from its intracellular binding sites. Comparison with other cell types showed that the extent of intracellular Li(+)/Mg(2+) competition at the same Li(+) loading level depends on intracellular Li(+) transport and immobilisation in a cell-specific manner, being maximal for neuroblastoma cells. PMID- 15110990 TI - Effects of autolysis on properties of mu- and m-calpain. AB - Although the biochemical changes that occur during autolysis of mu- and m-calpain are well characterized, there have been few studies on properties of the autolyzed calpain molecules themselves. The present study shows that both autolyzed mu- and m-calpain lose 50-55% of their proteolytic activity within 5 min during incubation at pH 7.5 in 300 mM or higher salt and at a slower rate in 100 mM salt. This loss of activity is not reversed by dialysis for 18 h against a low-ionic-strength buffer at pH 7.5. Proteolytic activity of the unautolyzed calpains is not affected by incubation for 45 min at ionic strengths up to 1000 mM. Size-exclusion chromatography shows that ionic strengths of 100 mM or above cause dissociation of the two subunits of autolyzed calpains and that the dissociated large subunits (76- or 78-kDa) aggregate to form dimers and trimers, which are proteolytically inactive. Hence, instability of autolyzed calpains is due to aggregation of dissociated heavy chains. Autolysis removes the N-terminal 19 (m-calpain) or 27 (mu-calpain) amino acids from the large subunit and approximately 90 amino acids from the N-terminus of the small subunit. These regions form contacts between the two subunits in unautolyzed calpains, and their removal leaves only contacts between domain IV in the large subunit and domain VI in the small subunit. Although many of these contacts are hydrophobic in nature, ionic-strength-induced dissociation of the two subunits in the autolyzed calpains indicates that salt bridges have an important, possibly indirect, role in the domain IV/domain VI interaction. PMID- 15110991 TI - The role of p27(Kip1) in maintaining the levels of D-type cyclins in vivo. AB - This in vivo study employs p27-deficient mice to investigate the significance of p27 for the metabolism of D-type cyclins in differentiated cells. The absence of p27 results in decreased levels of cyclins D2 and/or D3 in some organs. As demonstrated on Leydig cells of testis, such dependency is only restricted to certain cell types including terminally differentiated ones, and the absence of p27 in these cells can interfere with their differentiation. The decrease of cyclin D caused by the absence of p27 equals the amount of cyclin D physically associated with p27 in non-mutant animals. The data indicate that it is the proportion of p27-associated cyclin D that determines the response to p27 deficiency. Cells in which the level of D-type cyclin is dependent on p27 do not up-regulate the activity of their CDK2 and CDK4 upon loss of p27, and these cells have a negligible amount of p27 bound to CDK2 and/or cyclin A/E under normal conditions. Together, the findings suggest the existence of a dual role for p27, one being a classical regulation of cell cycle via inhibition of cyclin-dependent kinases (CDK), and the other being participation in the establishment and/or maintenance of differentiated status that is realized in conjunction with D-type cyclins. PMID- 15110992 TI - Tumour metabolites regulate tissue kallikrein in human umbilical vein endothelial cells. AB - Angiogenesis, the sprouting of new blood vessels, is tightly mediated via a myriad of endogenous factors. A pro-angiogenic alteration facilitates the formation of neovascular tumour networks, thereby providing mechanisms for uncontrolled growth. The kallikrein-kinin system is postulated to be pro angiogenic since its components have been detected in both endothelial cells and tumour tissue. No studies have, however, focussed on the role of tissue kallikrein (TK) in human angiogenic endothelial cell-tumour interactions. This study has optimised a challenge model whereby endothelial cells are presented with neuroblastoma metabolites, and vice versa. Image analysis of immunoreactive TK revealed a dose-dependant, significant reduction of TK localisation within endothelial cells, while gene expression remained unchanged, the latter determined by in situ RT-PCR. Neuroblastoma cells, when challenged with endothelial cell metabolites, displayed no change in TK synthesis or localisation. Alterations in TK synthesis and/or storage by angiogenic endothelial cells may be mediated by tumour-released signals and possibly indicate a shift from a proteolytic to a mitogenic function of TK. The challenge model provides a relatively simple experimental system to study angiogenic factors in tumour-endothelial cell interaction, and is the first to localise both TK and its mRNA within angiogenic endothelial and tumour cells. PMID- 15110994 TI - Brain-specific RGS9-2 is localized to the nucleus via its unique proline-rich domain. AB - Brain-specific regulator of G protein signaling 9 (RGS9-2) is a member of a family of proteins that can function as GTPase-activating proteins for heterotrimeric G proteins. In the present study, we examined the intracellular distribution of RGS9-2 in native brain tissue and transfected cells. Immunocytochemical and immunoblot experiments revealed an unexpectedly high proportion of RGS9-2 within the nuclei of forebrain neurons. A similar intracellular distribution was seen in transfected COS-7 cells. The RGS9 binding partner G(beta5) further enhanced the nuclear localization of RGS9-2, but did not affect the strongly cytoplasmic localization of RGS9-1, the retinal form of RGS9. Deletion construct analysis revealed that the unique polyproline-rich C-terminus of brain-specific RGS9-2 contains sequences necessary and sufficient to target RGS9 to the nucleus of COS-7 cells, as well as cultured striatal neurons. Furthermore, RGS9-2 transfection increased the transcriptional activity of a neuronal gene construct normally expressed in RGS9-positive neurons, suggesting that nuclear RGS9 directly or indirectly regulates transcription in vivo. The nuclear localization of RGS9-2 suggests a heretofore-unanticipated role for this brain-specific protein in transducing signals to the nuclei of forebrain neurons. PMID- 15110996 TI - Alcohols increase calmodulin affinity for Ca2+ and decrease target affinity for calmodulin. AB - It has been proposed that alcohols and anesthetics selectively inhibit proteins containing easily disrupted motifs, e.g., alpha-helices. In this study, the calcineurin/calmodulin/Ca(2+) enzyme system was used to examine the effects of alcohols on calmodulin, a protein with a predominantly alpha-helical structure. Calcineurin phosphatase activity and Ca(2+) binding were monitored as indicators of calmodulin function. Alcohols inhibited enzyme activity in a concentration dependent manner, with two-, four- and five-carbon n-alcohols exhibiting similar leftward shifts in the inhibition curves for calmodulin-dependent and independent activities; the former was slightly more sensitive than the latter. Ca(2+) binding was measured by flow dialysis as a direct measure of calmodulin function, whereas, with the addition of a binding domain peptide, measured calmodulin-target interactions. Ethanol increased the affinity of calmodulin for Ca(2+) in the presence and absence of the peptide, indicating that ethanol stabilizes the Ca(2+) bound form of calmodulin. An increase in Ca(2+) affinity was detected in a calmodulin binding assay, but the affinity of calmodulin for calcineurin decreased at saturating Ca(2+). These data demonstrate that although specific regions within proteins may be more sensitive to alcohols and anesthetics, the presence of alpha-helices is unlikely to be a reliable indicator of alcohol or anesthetic potency. PMID- 15110995 TI - Induction of ferritin expression by oxalomalate. AB - Ferritin is a ubiquitous protein required for intracellular iron storage; its biosynthesis is mainly regulated by iron-regulatory proteins (IRP1 and IRP2) at post-transcriptional level. This regulation prevents iron excess from promoting the formation of reactive oxygen species (ROS). IRP1 is regulated by such factors as intracellular iron levels, the oxidants H(2)O(2) and NO. We recently demonstrated that oxalomalate (OMA, alpha-hydroxy-beta-oxalosuccinic acid), a competitive inhibitor of aconitase, which is an enzyme of the citric acid cycle, remarkably decreases the binding activity of IRP1. The aim of the present study was to investigate whether this molecule could affect the expression of ferritin. The RNA-binding activity of IRP1, evaluated by gel retardation assay, decreased after treatment of several cell lines with 5 mM OMA, with a maximal decrease of about 3-fold after 6 h. This effect remained almost constant up to 48 h after which it returned to basal levels. Intracellular ferritin levels, determined by Western blot analysis, increased in correlation with the OMA-induced decrease of IRP1 binding activity. Furthermore, treatment of cells with OMA caused a rise in ferritin mRNA levels. Interestingly, in cells exposed to iron challenge, OMA induced overexpression of ferritin prevented formation of ROS and cellular lipid peroxidation. These data show that an inhibitor of aconitase, OMA, besides being involved in energetic metabolism, is able to control ferritin expression, probably through molecular mechanisms of either post-transcriptional regulation or transcriptional modulation, with advantageous consequences for the cell. PMID- 15110993 TI - Alpha-chemokine-mediated signal transduction in human Kaposi's sarcoma spindle cells. AB - The role of chemokines and their receptors in HIV biology and Kaposi's sarcoma (KS) pathogenesis has recently gained considerable attention. It has been shown that KS-associated human herpes virus type 8 (KSHV/HHV-8) encodes functional homologues of certain chemokines and chemokine receptors. This suggests that chemokines may contribute to the growth and spread of KS seen in AIDS. We found the expression of CXCR4 in primary KS tissue by using in situ hybridization (ISH). Recently, alpha-chemokine receptors CXCR1 and CXCR2 have also been shown to be expressed by KS tissues. We further characterized the expression of these chemokines as well as the signaling events induced upon binding to their respective cognate ligands in the KS 38 spindle cell line. These cells express authentic characteristics of primary KS spindle cells and provide a useful in vitro model for these studies. We observed using RT-PCR that KS 38 cells express mRNA for the alpha-chemokine receptors CXCR1, CXCR2, and CXCR4. We also confirmed the cell surface protein expression by FACS analysis. Characterization of signaling pathways revealed that the alpha-chemokines, IL-8 and stromal cell derived factor 1alpha (SDF1alpha/CXCL12), activated members of the mitogen activated protein (MAP) kinase family, including Erk kinase, c-Jun amino terminal kinase (JNK)/stress-activated protein kinase (SAPK) and the p38 MAP kinase. Furthermore, using DNA protein-binding experiments, we have shown that IL-8 increased AP-1 and NF Kappa B activity in these cells. IL-8 also enhanced the chemotaxis of KS cells. These results reveal that chemokine-induced signaling pathways may mediate cell growth, transcriptional activation and cell migration in KS. PMID- 15110997 TI - Expression of functional beta2-adrenergic receptors in the lung epithelial cell lines 16HBE14o(-), Calu-3 and A549. AB - Adrenergic drugs acting through the beta(2)-adrenoceptor (beta(2)-AR) adenylate cyclase (AC) signal transduction system elicit a variety of responses within the mammalian airway epithelium; however, its composition of multiple phenotypically differentiated cell types complicates the understanding of the regulation cascades within this tissue. The present study evaluates beta(2)-AR mRNA level, number, subtype and the cyclic adenosine-3',5'-monophosphate (cyclic AMP) response to isoproterenol (iso) in the human airway epithelial cell lines 16HBE14o(-), Calu-3 and A549, using reverse transcriptase polymerase chain reaction (RT-PCR), radioligand binding studies, [(3)H]-radioimmunoassay and immunocytochemical staining. After 4-5 days in culture, all three cell types produced beta(2)-AR mRNA and protein at a magnitude of gene expression levels Calu-3>or=16HBE14o(-)>A549, whereas control cells Cos-1 and Caco-2 were negative. The beta(2)-AR adenylate cyclase system was highly expressed and functional in the human airway epithelial cells Calu-3 and 16HBE14o(-). The mean beta(2)-AR density (B(max)), equilibrium dissociation constant (K(D)), and the percentage of beta-AR subtypes assessed by radioligand binding were approximately 9908+/-1127 and 6423+/-895 binding sites/cell, 32+/-2.7 pM and 25+/-1.1 pM, and approximately 100% in Calu-3 and 16HBE14o(-)cells, respectively. However, in the alveolar cell type A549 the cell surface beta(2)-AR was virtually undetectable by (-)-[(125)I] iodocyanopindolol (ICYP) binding. Stimulation of cultured cells with (-) isoproterenol enhanced the basal cyclic AMP accumulation only in Calu-3 and 16HBE14o(-) cells, which was blocked by the beta(2)-selective antagonist ICI 118,551, but not by the beta(1)-selective antagonist CGP 20712A, confirming functional coupling of the beta(2)-AR to adenylate cyclase in these cells. Immunocytochemical staining localised the receptor on the cell membrane and the cytoplasm in Calu-3 and 16HBE14o(-) cells, while it was confined to the cytoplasm only in A549 cells. In conclusion, the beta(2)-AR expression and its functional coupling to adenylyl cyclase was very high in the human airway epithelial cells Calu-3 and 16HBE14o(-), but not in A549, suggesting that the cell lines Calu-3 and 16HBE14o(-) present suitable models to study function and regulation of the beta-adrenoceptor signalling in the respiratory system. PMID- 15110998 TI - Amphibian transition to the oxidant terrestrial environment affects the expression of glutathione S-transferases isoenzymatic pattern. AB - It has been postulated that glutathione S-transferases (GST; EC 2.5.1.18) may play a role in protecting against oxidative stress. In previous studies, we have purified and characterised from Bufo bufo embryos a GST isoenzyme (BbGSTP1-1), which falls at very low level in the adult liver, where a novel isoform (BbGSTP2 2), starts to be highly expressed. During transition to adult life, B. bufo leaves the aquatic environment to live predominantly in the terrestrial environment, characterised by higher oxygen concentration. It has been found that BbGSTP2-2 is more efficient in scavenging from organic hydroperoxides. Therefore, the appearance of BbGSTP2-2 may respond to the necessity of providing the adult toad with a more suitable protection against oxygen toxic by-products. In this work, we performed experiments aimed at verifying if oxidative stress (hyperoxic and H(2)O(2) treatments) could act as a modulator of BbGSTP2-2 expression. Results show that: (a) BbGSTP2 mRNA starts to be expressed in the late embryonic period, while protein appears during metamorphosis; (b) oxidative stress induces anticipation of BbGSTP2 gene expression at both transcriptional and translational levels. These findings seem to indicate that the appearance of BbGSTP2-2 is aimed at endowing the adult toad with more efficient antioxidant defence in the terrestrial atmosphere. PMID- 15110999 TI - Modal gating of NMDA receptors. AB - The decay time course of excitatory postsynaptic currents generated by slow glutamatergic synapses is determined by the single-channel kinetics of postsynaptic NMDA receptor channels. In a recent study, examination of these single-channel kinetics has revealed that NMDA receptors can enter into modal gating. Linear free-energy analysis suggests that the various modes arise from a common reaction mechanism, providing further insight into the gating underlying decay of the slow synaptic response. PMID- 15111000 TI - RNA gains a new function: a mediator of neurodegeneration. AB - Over the past couple of years, it has become evident that some males carrying the fragile X mental retardation (FMR1) premutation, typically viewed to be normal, in fact manifest a neurodegenerative disease with ataxia and tremor. FMR1 premutation alleles uniquely produce FMR1 transcripts with an elongated CGG repeat, leading to the hypothesis that premutant transcripts cause the neurodegenerative disease in carriers. Recently Jin et al. demonstrated, in Drosophila, that FMR1 premutation RNA causes neurodegeneration. These data show RNA can induce neurodegeneration and provide strong evidence that FMR1 RNA mediates the neurodegeneration in human premutation carriers. PMID- 15111001 TI - Gene therapy for ALS delivers. AB - Amyotrophic lateral sclerosis (ALS) is a fatal, progressive neurodegenerative disease that kills motor neurons. Despite a long disappointing history of human trials with neurotrophins, including insulin-like growth factor 1 (IGF-1), Kaspar and colleagues have successfully slowed disease in transgenic ALS mice by forcing motor neurons to produce IGF-1 following retrograde delivery of recombinant adeno associated virus (AAV) injected into muscle. With the clinical safety of both IGF 1 and AAV already established, this provides real hope for an effective treatment of ALS. PMID- 15111002 TI - Star-cross'd neurons: astroglial effects on neural repair in the adult mammalian CNS. AB - Astroglia have long been thought to play merely a supporting role in the life of the neuron. However, these star-shaped cells have recently been the focus of intense study that has begun to emphasize remarkable and novel roles for these amazing cells. While astroglia play positive roles in the life of the neuron, they can simultaneously exert negative influences. Kinouchi et al. convincingly demonstrate and characterize an inhibitory role played by astroglia after neuronal transplantation. These findings remind us that astroglia exert positive and negative influences on neuronal survival, migration, neurite outgrowth and functional integration. Here, we review the complementary and often contradictory roles of astroglia during neuronal integration. PMID- 15111004 TI - Response to Quinlan and Nilsson: Astroglia sitting at the controls? PMID- 15111005 TI - Do acetylcholinesterase inhibitors boost synaptic scaling in Alzheimer's disease? AB - Studies on neural networks indicate that the pattern of amnesia that occurs in Alzheimer's disease (AD) can be reproduced by synaptic loss, but only when this loss is accompanied by synaptic scaling, a homeostatic mechanism that maintains the level of excitatory input on postsynaptic neurons. It is suggested that increased cholinergic activity during the early stages of AD contributes to synaptic scaling and that acetylcholinesterase inhibitors improve cognition in AD patients by boosting this mechanism. PMID- 15111003 TI - Reloading the retina by modifying the glial matrix. AB - Remarkably, removing the major structural elements from astrocytes and Muller glia in the mouse retina makes successful integration and differentiation of stem cells into the retina more likely, as revealed by Kinouchi et al. These ground breaking results throw up several new conundrums in understanding the process of stem cell recruitment, differentiation and incorporation into the retina, but they also suggest novel ways to treat retinal degenerative diseases. PMID- 15111007 TI - It's green outside: tracking cell surface proteins with pH-sensitive GFP. AB - Green fluorescent protein (GFP) and mutated GFP variants have proved to be immensely powerful tools that have had a profound impact on research in biological sciences. This review considers the development, use and future implications of pH-dependent GFP variants (e.g. pHluorins). These proteins hold considerable promise for the relatively non-invasive monitoring of events such as exocytosis, endocytosis and protein surface expression in living neurons with high spatial and temporal resolution. PMID- 15111008 TI - Tonically active GABA A receptors: modulating gain and maintaining the tone. AB - GABAA receptors not only respond to the local release of GABA from presynaptic terminals, but can also mediate a persistent 'tonic current'. This reflects the activation of high-affinity GABAA receptors by ambient GABA concentrations. Tonic GABAA-receptor-mediated signalling occurs in different brain regions, shows cell type-specific differences in magnitude and pharmacology, and changes during brain development. Some clues to the adaptive significance of this phenomenon are beginning to emerge: in cerebellar granule cells, it alters the gain of transmission of rate-coded sensory information; in the hippocampus, it acts in a cell-type-specific manner to regulate the excitability of the network. Because tonic conductances can be modulated by changes in GABA release and uptake, and by modulators of high-affinity GABAA receptors including neurosteroids, this phenomenon provides a potentially important new window onto neuronal information processing and pathological states such as epilepsy. PMID- 15111009 TI - DAncing past the DAT at a DA synapse. AB - Spillover of dopamine (DA) from a release site into the extrasynaptic space is widely acknowledged. Indeed, spillover is necessary for signalling by DA because its receptors are predominantly extrasynaptic. Dopamine transporters (DATs) are often considered to participate in this process by 'gating' spillover. This article reviews the competition between DATs and diffusion in sculpting extracellular DA transients after quantal release, using a model based on data from the literature. Its conclusions challenge the view that DATs limit synaptic DA concentration and gate initial spillover from a release site; this is the work of diffusion. Rather, the greatest influence of DATs, or of their inhibition, is on the sphere of influence and lifetime of DA beyond a release site and, thus, on net extracellular concentration. PMID- 15111006 TI - Memory traces of trace memories: neurogenesis, synaptogenesis and awareness. AB - To associate events that are disparate in time, the brain must record, retain and perhaps even reflect on the individual events themselves. Aspects of such learning can be probed with trace conditioning, during which an animal learns to associate events that are temporally distant from one another. For decades, we have known that the formation of so-called trace memories (in which one stimulus is associated with a second stimulus that is discontinuous and later in time) depends on the hippocampal formation. Recent findings indicate that the hippocampus is crucial for the initial acquisition of trace memories but not for their expression or long-term storage. More recent findings implicate neurogenesis, synaptogenesis and awareness in the formation of trace memories. PMID- 15111010 TI - Neural mechanisms for prediction: do insects have forward models? AB - 'Forward models' are increasingly recognized as a crucial explanatory concept in vertebrate motor control. The essential idea is that an important function implemented by nervous systems is prediction of the sensory consequences of action. This is often associated with higher cognitive capabilities; however, many of the purposes forward models are thought to have analogues in insect behaviour, and the concept is closely connected to those of 'efference copy' and 'corollary discharge'. This article considers recent evidence from invertebrates that demonstrates the predictive modulation of sensory processes by motor output, and discusses to what extent this supports the conclusion that insect nervous systems also implement forward models. Several promising directions for further research are outlined. PMID- 15111011 TI - Targeting eNOS for stroke protection. AB - Nitric oxide (NO) generated by endothelial NO synthase (eNOS) plays a crucial role in vascular function and homeostasis. NO possesses vasodilatory, anti inflammatory, antithrombotic and antiproliferative properties. Augmentation of NO production increases cerebral blood flow, which can lead to neuroprotection during brain ischaemia. Several modalities that upregulate eNOS expression and/or activity have recently been identified, including HMG-CoA reductase inhibitors (statins), steroid hormones, nutrients and physical activity. They all increase NO bioavailability, leading to enhanced cerebral blood flow and protection from ischaemic stroke. Thus, therapeutic modalities that target eNOS not only serve as preventive measures to reduce stroke incidence but also could represent novel treatment strategies for reducing brain injury during cerebral ischaemia. PMID- 15111012 TI - Both mGluR1 and mGluR5 mediate Ca2+ release and inward currents in hippocampal CA1 pyramidal neurons. AB - Using combined whole-cell voltage-clamp recording and Ca2+ imaging we have investigated further the characteristics and pharmacology of group I metabotropic l-glutamate receptor (mGluR)-mediated responses in CA1 pyramidal neurons of the rat hippocampus. The selective group I mGluR agonist, (RS)-3,5 dihydroxyphenylglycine (DHPG), evoked a transient increase in intracellular Ca2+ levels ([Ca2+]i), within neuronal somas and apical dendrites, together with a relatively long lasting inward current (I(DHPG)). Both types of response were enhanced by depolarisation (-30 mV), and this condition was used for their characterisation. The DHPG-induced [Ca2+]i rise was much more sensitive to manipulations of Ca2+ homeostasis, such as using the Ca2+ store depleting agent, cyclopiazonic acid (50-100 microM), the fast Ca2+ buffer, BAPTA (intracellular; 20-40 mM) and Ca(2+)-free/EGTA (1 mM) bath solution, than I(DHPG), suggesting that these responses are, in the main part, mediated by distinct processes. The selective mGluR1 and mGluR5 antagonists, (S)-(+)-alpha-amino-a methylbenzeneacetic acid (LY367385; 100 microM) and 2-methyl-6-(phenylethynyl) pyridine (MPEP;10 microM), respectively, markedly inhibited both I(DHPG) and the DHPG-evoked increase in [Ca2+]i. Moreover, these antagonists inhibited the Ca2+ response by more than 50% suggesting a synergistic interaction between mGluR1 and mGluR5. This study demonstrates that in CA1 pyramidal neurons group I mGluR mediated inward currents and Ca2+ release from intracellular stores are enhanced under depolarising conditions and that mGluR1 and mGluR5 both contribute to these phenomena. PMID- 15111014 TI - GABAA and GABAB receptors in the nucleus accumbens shell differentially modulate dopamine and acetylcholine receptor-mediated turning behaviour. AB - The ability of GABAA and GABAB receptors in the shell of the nucleus accumbens to modulate distinct types of turning behaviour was investigated in freely moving rats, using the unilateral injection technique. The GABAA receptor agonist muscimol and the GABAA receptor antagonist bicuculline did not produce turning behaviour; the same holds for the GABAB agonist baclofen and the GABAB antagonist 2-hydroxysaclofen. A mixture of the dopamine D1 receptor agonist SKF 38393 and the dopamine D(2/3) receptor agonist quinpirole has been found to elicit contraversive pivoting, when injected into the shell. This pivoting was dose dependently inhibited by muscimol, and the inhibitory effect of muscimol was antagonised by bicuculline. Pivoting was also dose-dependently inhibited by baclofen; however, 2-hydroxysaclofen did not antagonise the inhibitory effect. The acetylcholine receptor agonist carbachol has been found to elicit contraversive circling, when injected into the shell. This carbachol-induced circling was inhibited by baclofen, and 2-hydroxysaclofen antagonised the inhibitory effect. Carbachol-induced circling was also partially inhibited by muscimol; however, the inhibitory effect of muscimol was not antagonised by bicuculline. It is concluded that mesolimbic GABAA receptors exert an inhibitory control on dopamine-dependent pivoting that can be elicited from the shell of the nucleus accumbens, and that GABAB receptors exert an inhibitory control on acetylcholine-dependent circling that can be elicited from the shell of the nucleus accumbens. This data extends the earlier reported findings that the neurochemical substrate in the shell of the nucleus accumbens that mediates dopamine-dependent pivoting is fundamentally different from the shell substrate that mediates acetylcholine-dependent circling. PMID- 15111013 TI - Enhanced dizocilpine efficacy in heterozygous reeler mice relates to GABA turnover downregulation. AB - Reelin synthesized by cortical GABAergic interneurons throughout the telencephalon is secreted into the extracellular matrix (ECM) and binds with nM affinity to integrin receptors located at dendritic spine postsynaptic densities and positively modulates Arc and other dendritic resident mRNAs translation, thereby facilitating the onset of synaptic plasticity and LTP consolidation. Accordingly, the reelin haploinsufficient heterozygous reeler mice (HRM) express a marked decrease of cortical thickness, of cortical and hippocampal dendritic spine density, and of cortical GAD67 expression. Behaviorally, HRM manifest a sensorimotor deficit, an exaggerated response to fear, and a deficit in olfactory discrimination learning. HRM and wild-type mice (WTM) were trained to retrieve to criterion palatable chocolate-flavored food pellets in an eight-arm radial maze. In 9-14 days of training HRM and WTM learned the task equally well committing only a few errors. However, HRM, when compared with WTM, show a greater cognitive impairment following the administration of dizocilpine. Also, HRM are more susceptible to the increased locomotion and stereotypic behavior elicited by dizolcipine. The enhanced dizocilpine susceptibility of HRM is not due to differences in pharmacokinetics because the levels of dizocilpine in cortices of HRM and WTM were virtually equal. We also failed to detect differences between HRM and WTM in glutamate brain content and in the rate of 13C-glucose incorporation into the glutamate brain pools. In contrast we found that the conversion index of glutamate into GABA (an indirect measurement of GABA turnover rate) is decreased in cortex, hippocampus and striatum of HRM when compared to WTM. Thus, HRM recapitulate several neurochemical and behavioral endophenotypes reminiscent of schizophrenia and these mice can be proposed as a relevant animal model for the study of pharmacological treatments aimed at alleviating the sensory-motor and cognitive dysregulation associated with schizophrenia. PMID- 15111015 TI - Role of mu- and delta-opioid receptors in the nucleus accumbens in turning behaviour of rats. AB - The role of mu-, delta1- and delta2-opioid receptors in the nucleus accumbens in pivoting was investigated in freely moving rats. Unilateral injections of the mu opioid receptor agonist, [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO, 1 and 2 microg) and the delta2-opioid receptor agonist, deltorphin II (1 and 2 microg), but not the delta1-opioid receptor agonist, [D-Pen(2,5)]-enkephalin (DPDPE, 1-4 microg), into the shell or the core of the nucleus accumbens significantly induced contraversive pivoting. The pivoting induced by DAMGO (2 microg) and deltorphin II (2 microg) was inhibited significantly by the mu-opioid receptor antagonist, D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Phe-Thr-NH2 (CTOP, 0.1 and 1 microg), and the delta2-opioid receptor antagonist, naltriben (NTB, 0.1 and 1 mg/kg, i.p.), respectively. The DAMGO (2 microg)- or deltorphin II (2 microg)-induced pivoting was also inhibited significantly by co-administration of the dopamine D1/D2 receptor antagonist, cis(Z)-flupentixol (1 and 10 microg). The pivoting induced by unilateral injections of a mixture of dopamine D1 (SKF 38393, 5 microg) and D2 (quinpirole, 10 microg) receptor agonists into the shell was significantly inhibited by cis(Z)-flupentixol (1 and 10 microg) or NTB (1 and 3 mg/kg, i.p.), but not CTOP (1 microg) or delta1-opioid receptor antagonist, (E)-7 benzylidenenaltrexone (1 mg/kg, i.p.). The contraversive pivoting elicited by the cholinergic agonist, carbachol (5 microg), into the core was inhibited by co administration of the muscarinic M1 antagonist, pirenzepine (1 microg), but not cis(Z)-flupentixol (1 microg). The results suggest that unilateral activation of mu- or delta2-opioid, but not delta1-opioid, receptors in the core and/or shell of the nucleus accumbens elicits contraversive pivoting that requires intact dopamine D1/D2 receptors in the shell, but not intact muscarinic M1 mechanism in the core. The study also shows that delta2-opioid, but not mu- and delta1-opioid, receptors in the core and/or shell modulate the shell-specific, dopamine D1/D2 receptor mechanisms involved in the production of pivoting. PMID- 15111016 TI - Topiramate selectively protects against seizures induced by ATPA, a GluR5 kainate receptor agonist. AB - Although the mechanism of action of topiramate is not fully understood, its anticonvulsant properties may result, at least in part, from an interaction with AMPA/kainate receptors. We have recently shown that topiramate selectively inhibits postsynaptic responses mediated by GluR5 kainate receptors. To determine if this action of topiramate is relevant to the anticonvulsant effects of the drug in vivo, we determined the protective activity of topiramate against seizures induced by intravenous infusion of various ionotropic glutamate receptor agonists in mice. Topiramate (25-100 mg/kg, i.p.) produced a dose-dependent elevation in the threshold for clonic seizures induced by infusion of ATPA, a selective agonist of GluR5 kainate receptors. Topiramate was less effective in protecting against clonic seizures induced by kainate, a mixed agonist of AMPA and kainate receptors. Topiramate did not affect clonic seizures induced by AMPA or NMDA. In contrast, the thresholds for tonic seizures induced by higher doses of these various glutamate receptor agonists were all elevated by topiramate. Unlike topiramate, carbamazepine elevated the threshold for AMPA- but not ATPA induced clonic seizures. Our results are consistent with the possibility that the effects of topiramate on clonic seizure activity are due to functional blockade of GluR5 kainate receptors. Protection from tonic seizures may be mediated by other actions of the drug. Together with our in vitro cellular electrophysiological results, the present observations strongly support a unique mechanism of action of topiramate, which involves GluR5 kainate receptors. PMID- 15111017 TI - IDRA-21, a positive AMPA receptor modulator, inhibits synaptic and extrasynaptic NMDA receptor mediated events in cultured cerebellar granule cells. AB - IDRA-21 (7-chloro-3-methyl-3,4-dihydro-2H-1,2,4-benzothiadiazine 1,1-dioxide) reduces alpha-amino-3-hydroxy-5-methylisoxazolepropionic acid (AMPA) receptors desensitisation in vitro and restores learning and cognitive impairment in vivo. In this study, we show that in cerebellar granule cells (CGCs) in culture IDRA-21 reduces N-methyl-d-aspartate receptor (NMDAR) whole-cell currents. The effect is neither competitive nor voltage-dependent. The reduction of NMDA currents is stronger at low glycine concentrations suggesting an interaction with this site. IDRA-21 shortens miniature excitatory postsynaptic currents mediated by NMDARs (NMDA-mEPSCs) in CGCs grown in low potassium with no effect on peak amplitudes. By using fast glutamate application onto CGCs nucleated patches, we found that IDRA-21 decreases both decay time constant and amplitude of the current. Experiments performed on recombinant NMDAR expressed in HEK 293 cells showed that IDRA-21 was more effective on NR1a-NR2B than NR1a-NR2A receptors highlighting a subunit selectivity of the drug. Our findings make light on a novel target for IDRA-21: NMDA receptors function is negatively modulated and the different action at the level of extrasynaptic and synaptic receptors could be ascribed to a partial selectivity for NR2B subunits. PMID- 15111018 TI - Cloning and characterization of a novel G-protein-coupled receptor with homology to galanin receptors. AB - We report the identification, cloning, and localization of human and mouse orthologues of a new G-protein-coupled receptor with homology to galanin receptors, which we termed galanin-receptor like (GalRL). The genes of GalRL were localized to chromosome 5q32 in human and to 18B3 in mouse. Northern blot analysis revealed expression of GalRL in the central nervous system of human and mouse and in 7- and 15-day-old mouse embryos. Minor levels were found in some peripheral organs and tissues, such as testis, liver, kidney and stomach. In situ hybridization experiments demonstrated predominant expression of GalRL in the central nervous system, with a distinct localization in the habenular complex. In the peripheral nervous system single neurons of sensory ganglia were labeled. During embryonal development the expression was more widespread in the nervous system, where in addition to the dorsal thalamus, hybridization signals were detected in other areas of the brain including the striatum, the locus coeruleus, and several hindbrain nuclei. A weak activation of GalRL by galanin suggests that the endogenous ligand shares structural features with galanin. PMID- 15111019 TI - SSR181507, a putative atypical antipsychotic with dopamine D2 antagonist and 5 HT1A agonist activities: improvement of social interaction deficits induced by phencyclidine in rats. AB - Social behaviour is frequently impaired in schizophrenic patients, and current antipsychotics appear poorly effective in alleviating this deficit. SSR181507 is a selective dopamine D2 receptor antagonist and 5-HT1A receptor agonist [Neuropsychopharmacology 28 (2003) 2064] with an atypical antipsychotic profile and additional antidepressant/anxiolytic activities [Neuropsychopharmacology 28 (2003) 1889]. Here, we sought to assess the efficacy of SSR181507, and of reference antipsychotics and antidepressant/anxiolytics, to counteract phencyclidine (PCP)-induced social interaction deficit in rats. Pairs of unfamiliar rats were placed for 10 min each day into a dimly lit arena, during four consecutive days. On the test day (5th day), each pair was placed into the arena 30 min after i.p. treatment with PCP (or vehicle) and a challenge compound or vehicle (same for both rats, i.p. or s.c.). The time spent in social interaction was scored during 10 min. PCP (1 mg/kg) decreased social interaction time by about 35%. This effect was fully antagonized by pre-treatment with SSR181507 (1 mg/kg). In contrast, neither haloperidol (0.05 and 0.1 mg/kg) nor clozapine (0.3 and 1 mg/kg) antagonized this PCP-induced deficit. The selective 5 HT1A receptor agonist 8-OH-DPAT (0.025 and 0.05 mg/kg s.c.), but not the anxiolytic diazepam (0.75 and 1.5 mg/kg), also improved social interaction impairment in PCP-treated rats: this would indicate that the 5-HT1A receptor agonist properties of SSR181507 are responsible for the reversal of PCP-induced social deficit. These data suggest that, in addition to its atypical antipsychotic profile and antidepressant/anxiolytic activities, SSR181507 has a potential therapeutic activity in another key feature of schizophrenia poorly controlled by current antipsychotics, namely deterioration in social functioning. PMID- 15111020 TI - Prevention of MPTP (N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) dopaminergic neurotoxicity in mice by chronic lithium: involvements of Bcl-2 and Bax. AB - Lithium has been reported to exert neuroprotective activity in several neuronal cell cultures and in vivo models against glutamate toxicity. Since this action was reported to be associated with alterations in the antiapoptotic Bcl-2 family proteins, the effect of chronic lithium diet on the ability of the parkinsonism neurotoxin, N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to deplete striatal dopamine in mice was determined. Mice were fed for with a diet containing 1.1, 2.2, 3.3, and 4.4 g/kg lithium chloride (LiCl) for 4 weeks, during which time serum levels of lithium were monitored. The 3.3 g/kg lithium diet gave serum level value very similar to what is observed in lithium therapy in man and the 4.4 g/kg well above this. At the end of this period the mice received 24 mg/kg MPTP i.p. once daily for 3 days. A direct relation was established with the increase in serum lithium and its ability to prevent MPTP induced depletion of striatal dopamine (DA) and its metabolites DPOAC and HVA. With the diet containing the highest lithium concentration there was an almost complete prevention of striatal dopamine depletion and the reduction in tyrosine hydroxylase activity and protein and it prevented the increase in dopamine turnover (DOPAC + HVA/DA) normally observed in MPTP treatment. Lithium did not interfere with the metabolism of MPTP, or with its brain uptake, since, the level of its monoamine oxidase (MAO) B derived metabolite, MPP+, in the striata of lithium and non-lithium treated mice were almost identical. Striatal Bcl-2 was significantly decreased, while Bax was increased in MPTP treated mice. Lithium treatment not only increased striatal Bcl-2 in control mice, but also prevented its reduction as induced by MPTP, and an opposing effect was seen with Bax. The neuroprotective action of lithium in this model of Parkinson's disease has been attributed to its antiapoptotic activity which among other factors includes induction of Bcl-2 and reduction of Bax. PMID- 15111021 TI - Subsets of acetylcholine-stimulated 86Rb+ efflux and [125I]-epibatidine binding sites in C57BL/6 mouse brain are differentially affected by chronic nicotine treatment. AB - Nicotinic cholinergic receptor (nAChR) sites that bind nicotine with high affinity (likely alpha4beta2-nAChR) increase following chronic nicotine treatment. Effects of chronic treatment on other nAChR binding sites and functional responses of nAChRs are less well studied. Therefore, C57BL/6 mice were intravenously infused for 10 days with saline or nicotine (five doses, 0.25 4.0 mg/kg/h) and nAChR function and three different nicotinic binding sites in 12 brain regions were assessed. Plasma nicotine and cotinine increased linearly with dose. 86Rb+ efflux with higher sensitivity to acetylcholine tended to decrease with increasing dose, whereas efflux with lower sensitivity to acetylcholine tended to increase. As anticipated, likely alpha4beta2-nAChR [125I]-epibatidine binding sites increased with treatment (estimated dosage for one-half maximal increase was 0.44 mg/kg/h, plasma nicotine approximately 20 ng/ml). 86Rb+ efflux with higher sensitivity to acetylcholine and cytisine-sensitive [125I] epibatidine binding are predominantly alpha4beta2-nAChR. A high correlation between these parameters was observed across brain regions and slopes of these regression lines decreased with treatment dose, suggesting a decrease in function per unit receptor. Likely alpha3beta4-nAChR binding sites were unaffected even at the highest dose (4.0 mg/kg/h, approximately 210 ng/ml). A third set of diverse nAChR binding sites increased in some brain regions, but only after high-dose treatment. PMID- 15111022 TI - Melatonin MT(1/2) receptor stimulation reduces cortical overflow of cholecystokinin-like material in a model of anticipation of social defeat in the rat. AB - The involvement of cholecystokinin (CCK) in the potential anxiolytic-like effects of melatonin and of the antitumor MT(1/2) receptor agonist, S23478, was assessed by measuring the cortical outflow of CCK-like material (CCKLM) in a rat model of anticipation of social defeat. After repeated social defeats by a male Tryon Maze Dull (TMD) rat, Sprague-Dawley (SD) rats were implanted for microdialysis in the frontal cortex and placed in the same environment as for the defeated sessions, but no confrontation with the TMD rat was allowed. Anticipation of social defeat induced anxiety-like behaviors (immobility, ultrasonic vocalization, defensive postures) associated with a significant increase (approximately +90%) in cortical CCKLM outflow in SD rats. Acute pretreatment with melatonin (5 or 40 mg/kg i.p.) or S23478, at 5 mg/kg i.p., had no or only minor effects on anxiety-like behaviors and did not affect CCKLM overflow. In contrast, at 40 mg/kg i.p., S23478 significantly reduced the duration of immobility and vocalization as well as the cortical CCKLM overflow (-30%) in defeated SD rats, and both effects were prevented by the MT(1/2) receptor antagonist S22153 (40 mg/kg i.p.). These data indicated that MT(1/2) receptor stimulation can exert anxiolytic-like effects associated with inhibition of cortical CCKergic neurotransmission in rats anticipating social defeat. PMID- 15111023 TI - Pharmacological characterisation of the orexin receptor subtype mediating postsynaptic excitation in the rat dorsal raphe nucleus. AB - Electrophysiological recordings from dorsal raphe nucleus (DRN) neurones in rat brain slices have revealed that the orexins can cause direct and reversible depolarisation of the postsynaptic membrane. Whilst it is known that the membrane depolarisation produced by orexin-A can dramatically increase the firing rate of DRN neurones, quantitative pharmacological analysis that determines the receptor subtype mediating the orexinergic response has not yet been performed. Here, we demonstrate that the rank order of potencies of orexin receptor agonists to excite serotonergic DRN neurones is orexin-A = orexin-B > SB-668875-DM. In contrast, the rank order of potency of these agonists to excite noradrenergic locus coreleus (LC) neurones is orexin-A > orexin-B > SB-668875-DM. We show further that the orexin receptor antagonist, SB-334867-A, inhibits the effects of orexin-A in the LC and DRN with pKB values of 6.93 and 5.84, respectively, values similar to those calculated for human OX1 (7.27) and OX2 (5.60) receptors expressed in CHO cells. These data suggest a differential role for OX1 and OX2 receptors in stimulating distinct populations of monoaminergic neurones in the rat CNS with OX2 receptors exhibiting a more pronounced functional significance in serotonergic neurones and OX1 in noradrenergic neurones. PMID- 15111024 TI - Evaluation of the effect of chronic antidepressant treatment on neurokinin-1 receptor expression in the rat brain. AB - Clinically effective antidepressants are thought to exert their therapeutic effects by facilitating central monoamine neurotransmission. However, recent data showing that neurokinin-1 receptor (NK1R) antagonists have antidepressant properties in both animal and clinical studies raise the possibility that classical antidepressants may also influence NK1R expression in the brain. To test this hypothesis, rats were treated with desipramine, paroxetine, venlafaxine, tranylcypromine or vehicle for 14-42 days. NK1R binding sites and mRNA were determined in a wide variety of brain areas using in situ hybridization and quantitative receptor autoradiography. In all areas examined, the abundance of NK1R binding sites was unchanged after 14 days of treatment. None of the treatments altered the number of NK1R binding sites following 42 days treatment with the exception that an increase was found in the locus coeruleus with tranylcypromine. Taken together, we report that repeated treatment with antidepressants of different classes does not cause significant changes in NK1R expression. PMID- 15111025 TI - Differential contributions of NOS isoforms in the rostral ventrolateral medulla to cardiovascular responses associated with mevinphos intoxication in the rat. AB - The organophosphate poison mevinphos (Mev) elicits cardiovascular responses via nitric oxide (NO) produced on activation of M2 muscarinic receptors (M2R) in the rostral ventrolateral medulla (RVLM), where sympathetic vasomotor tone originates. This study further evaluated the contribution of nitric oxide synthase (NOS) isoforms at the RVLM to this process, using adult Sprague-Dawley rats. Bilateral co-microinjection into the RVLM of the selective NOS I inhibitor (250 pmol), 7-nitroindazole or N(omega)-propyl-L-arginine antagonized the initial sympathoexcitatory cardiovascular responses to Mev (10 nmol). Co-administration of a selective NOS II inhibitor, N6-(1-iminoethyl)-L-lysine (250 or 500 pmol) further enhanced these cardiovascular responses and reversed the secondary sympathoinhibitory actions of Mev. A potent NOS III inhibitor, N5-(1-iminoethyl) L-ornithine (46 or 92 nmol) was ineffective. We also found that M2R co-localized only with NOS I- or NOS II-immunoreactive RVLM neurons. Furthermore, only NOS I or II in the ventrolateral medulla exhibited an elevation in mRNA or protein levels during the sympathoexcitatory phase, with further up-regulated synthesis of NOS II during the sympathoinhibitory phase of Mev intoxication. We conclude that whereas NOS III is not engaged, NO produced by NOS I and II in the RVLM plays, respectively, a sympathoexcitatory and sympathoinhibitory role in the cardiovascular responses during Mev intoxication. PMID- 15111026 TI - Involvement of arachidonic acid cascade in working memory impairment induced by interleukin-1 beta. AB - The aim of the present study is to clarify the possible relevance of the arachidonic acid cascade to working memory in rats, by using a three-panel runway apparatus. Interleukin-1 beta, injected bilaterally into the dorsal hippocampus at a dose of 100 ng/side, significantly impaired working memory, and this impairment was attenuated by pretreatment with 10 mg/kg (s.c.) of diclofenac, a cyclooxygenase inhibitor. Working memory was also impaired in rats administered a bilateral intrahippocampal injection of prostaglandin E2, in a dose-dependent manner at 0.01-1 microg/side. Furthermore, an injection of 100 ng/side of interleukin-1 beta significantly increased production of prostaglandin E2 (580 +/ 32 pg to 1142 +/- 101 pg/100 mg wet tissue) in the hippocampus. Taken together, these findings suggest that the activation of the arachidonic acid cascade was causative of the working memory impairment induced by interleukin-1 beta. PMID- 15111027 TI - Effects of isoflurane and ketamine on ATP-sensitive K channels in rat substantia nigra. AB - Whole cell recordings were made using midbrain slices to examine the effects of two different anaesthetics on ATP-sensitive K (K(ATP)) channels in principle neurons of rat substantia nigra pars compacta. When neurons were dialyzed with an ATP-free pipette solution during perfusion with a glucose-free external solution, a hyperpolarization and an outward current developed slowly in a tolbutamide inhibitable manner. The volatile anaesthetic 3% isoflurane slightly depolarised the neurons in the presence of ATP in the pipette solution and glucose in the external solution, but it did not affect the hyperpolarization or outward current in response to omission of ATP and glucose. Ketamine, an intravenous anaesthetic, did not change the membrane potential when ATP and glucose were included; however, it reversibly inhibited the hyperpolarization and outward current induced by intracellular ATP depletion in a dose-dependent manner. These effects of ketamine were not mimicked by AP-5, an NMDA receptor antagonist, or indatraline, an inhibitor of catecholamine uptake. These findings suggest that these anaesthetics have no stimulatory action on K(ATP) channels in these neurons when intracellular ATP is preserved and that ketamine but not isoflurane inhibits K(ATP) channels when the channels were activated by low intracellular ATP. PMID- 15111028 TI - Strategies of genetic biodiversity conservation in the marine environment. PMID- 15111029 TI - The application of histo-cytopathological biomarkers in marine pollution monitoring: a review. AB - During the past two decades, a variety of histopathological alterations in fish and bivalves have been developed and used as biomarkers in pollution monitoring. Some of these have been successfully adopted in major national monitoring programmes, while others, although show promise, are still in the experimental stage. This paper critically reviews the scientific basis, cause and effect relationship, reliability, advantages and limitations of 14 histo cytopathological biomarkers. The usefulness and practical application of each biomarker have been evaluated against a number of objective criteria including: ecological relevance, sensitivity, specificity, dose-response relationship, confounding factors, technical difficulties and cost-effectiveness. PMID- 15111030 TI - Vitellogenin induction as a biomarker of exposure to estrogenic compounds in aquatic environments. PMID- 15111031 TI - Seasonal and annual loads of hydrophobic organic contaminants from the Susquehanna River basin to the Chesapeake Bay. AB - Water from the Susquehanna River was collected and analyzed for polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyl (PCB) congeners to estimate seasonal and annual riverine loads to the Chesapeake Bay. Temporal variations in the chemical loads resulted from the large changes in the water flow rates and in the particle-associated contaminant concentrations. Concentrations of PCBs and PAHs in river particles (ng/g) were twice as great as those in the northern Chesapeake Bay, indicating that the Susquehanna River is an important source of these contaminants to the bay. The river carries a majority of its hydrophobic organic contaminants (HOCs) in the particulate phase. During periods of high flow, large amounts of suspended particles in the river result in elevated HOC levels and increased loadings of these contaminants to the bay. From 1997 to 1998, 60% of the total annual HOC loading occurred in the early spring coincident with high river flows. The total PCB and PAH annual loadings from the Susquehanna River to the Chesapeake Bay were 76 and 3160 kg/year, respectively and 75% of the loaded organic contaminants were in the particulate phase. Principal component analysis of PAH and PCB congener patterns in the particles reveals that the river suspended particles were dominated by autochthonous production in the summer and by resuspended sediment and watershed erosion during the winter and early spring. PMID- 15111032 TI - Water quality and health status of the Senegal River estuary. AB - The Senegal River estuary was sampled in May 2002 to get the first data on both the trophic and sanitary status of the water of the main river of the northwest African coast. Several physical, chemical and microbiological variables were measured twice along a transect. Inorganic nutrient concentrations were low while phytoplanktonic abundances (0.58-1.8 x 10(5) cells ml(-1)), bacterial abundances (0.27-8.1 x 10(7) cells ml(-1)), activity (22-474 pmol l(-1) h(-1)), were among the highest recorded in such ecosystems. Microbiological variables revealed a eutrophicated status for this estuary. Largest abundances of fecal contamination bacterial indicators were only detected in localized areas (Saint-Louis city and surrounding areas). The apparent good survival of fecal indicator bacteria in the estuarine waters despite a long residence time (4-5 days) has been evaluated by complementary survival experiments. Exposed to a salinity gradient, a local Escherichia coli strain showed a significantly better survival than those of an E. coli reference strain. PMID- 15111033 TI - Synergistic effect of sophorolipid and loess combination in harmful algal blooms mitigation. AB - The inhibition effect of sophorolipid and removal efficiency of loess on Cochlodinium polykrikoides and Alexandrium tamarense was investigated separately in the laboratory. Based on this, the combination of sophorolipid and loess for harmful algal bloom mitigation was proposed. Algal sedimentation tests in the laboratory and in the field revealed that the combination of sophorolipid and loess showed synergistic effects both on the removal efficiencies and on the mitigation cost. The concentration of 1 g/l loess and 5 mg/l sophorolipid was determined as the optimum ratio for C. polykrikoides mitigation. In the field test, the effective concentration of loess and sophorolipid in the combination group was reduced to 10% and 25%, respectively, compared to the non-combination group, and the cost decreased more than 60%. The combination of loess and sophorolipid was considered as a promising novel method in harmful algal bloom mitigation. PMID- 15111034 TI - Combining the use of gradients and reference areas to study bioaccumulation in wild oysters in the Hunter River estuary, New South Wales, Australia. AB - Two approaches for measuring the effects of human activities on aquatic biota are gradient studies and comparison of impacted areas to external references. Wild oysters were sampled at 12 sites adjacent to, upstream and downstream of a steelworks in the Hunter River estuary and in two reference estuaries. Regression analyses for the Hunter indicated reduced concentrations with distance from the point source for some metals and PAHs. Data compared from the reference estuaries to the two sites nearest and the two furthest from the point source in the Hunter indicated elevated concentrations of contaminants both near the point source and on an estuary-wide basis. The gradient approach was useful in identifying the industrial effluent as a point source for bioavailable chemicals. Combining this with the use of reference estuaries provided a broad geographic context in which to interpret results from the Hunter and identified estuary-wide effects. PMID- 15111036 TI - Indirect estimation of degradation time for zinc pyrithione and copper pyrithione in seawater. AB - The degradability of two antifouling biocides: zinc pyrithione (ZPT) and copper pyrithione (CPT) in seawater was examined. Reduction in toxicity due to degradation was monitored over two days using a bioassay with natural assemblages of coastal marine bacteria from Roskilde Fjord, Denmark. To investigate photo degradation of the compounds, bacteria were exposed to sterile ZPT- and CPT dilution that had either been exposed to sunlight or darkness. Bio-degradation was examined by diluting ZPT and CPT in sterile seawater or natural seawater. Photo-degradation half-life for ZPT was estimated to be 8.3+/-0.9 min and for CPT to 7.1+/-0.2 min. Total and microbial degradation in combination with photo degradation did not further shorten the degradation time, suggesting no bio degradation. Bio-degradation without the influence of sunlight was also negligible over the time-period investigated. ZPT and CPT are therefore suggested to persist in the marine environment where the influence of the light is limited. PMID- 15111035 TI - Distribution of contaminants in biota and sediments in the Musquash Estuary, Atlantic Canada, marine protected area site initiative and contaminant exclusion zone. AB - The Musquash Estuary, one of the last ecologically intact estuaries in New Brunswick, has been designated an area of interest for a marine protected area (MPA) under the Oceans Act. The area has been assessed for contaminant background levels as required for establishing MPA environmental quality. American lobster (Homarus americanus), blue mussel (Mytilus edulis) and sediments were collected for assessing contaminant levels and distribution in the harbour. Levels of contaminants from the indicator species and the abiotic component have shown: (1) two extremes of high and low Cu and Ag in lobster from the area; and (2) lower metal levels in inner Musquash Harbour sediments and mussels than in those from the harbour mouth. These suggest that deposition of contaminants into the Musquash MPA site was due to transport of contaminants by coastal currents from upstream coastal industrial activities. This reverse trend with higher contaminant levels in the biotic and abiotic components in the outer harbour than in the inner harbour differs from a contaminated harbour and suggests that a contaminant exclusion zone should be considered for controlling contamination from nearby coastal and estuarine industrial sites to protect the sensitive habitats within the marine protected area. PMID- 15111037 TI - Zinc concentrations in marine macroalgae and a lichen from western Ireland in relation to phylogenetic grouping, habitat and morphology. AB - Zinc concentrations in 19 species of marine macroalgae and a lichen from western Ireland (Spiddal, Co. Galway) were analysed by atomic absorption spectrophotometry. Algae were collected from a single site but occupied different shore levels and belonged to distinct phylogenetic groupings and to different (previously recognised) morphological groups. Concentrations ranged from 15-115 microg g(-1) dry weight. The greatest variation in concentration occurred amongst the red algal species, containing both maximum and minimum concentrations. Zn concentrations in brown and green algae were generally lower than those in red algae. When grouped according to thallus morphology, thin, branched sheets (which consisted mainly of red algae) contained the highest Zn concentrations. In filamentous algae, Zn levels were higher than in thick-leathery or coarsely branched algae. However, all green algal species examined had similar Zn concentrations, despite their different morphologies. In brown algae, the highest Zn levels were detected in mid-shore fucoids (Fucus vesiculosus and Fucus serratus), while thicker, subtidal and low-shore brown algae (Laminaria digitata, Halydris siliquosa) exhibited lower Zn concentrations. The lowest Zn concentrations were detected in high-intertidal species (Fucus spiralis, Pelvetia canaliculata), the only marine lichen examined (Ramalina siliquosa) and a red crustose alga (Corallina officinalis). In all morphological groups, red algal representatives contained relatively higher levels of Zn, the exception being Corallina officinalis. Zn levels in 4 species from a second, estuarine site in Galway Bay exhibited the same relative differences amongst species, but were all consistently higher than in algae from Spiddal. It is concluded that Zn accumulation in macroalgae is closely related to ecological growth strategies, following a functional-form model. However, the phylogenetic origin of species which determines carbohydrate and phenol composition, and responses to ambient environmental conditions ultimately determine, the availability of binding sites for polyvalent cations. PMID- 15111038 TI - Natural and anthropogenic hydrocarbons in sediments from the Chubut River (Patagonia, Argentina). AB - In March of 2001 a study was carried out to evaluate hydrocarbon levels in the lower course of the Chubut River. The study included 12 sample stations along the river from San Cristobal Bridge to the confluence with the sea, in a 25 km straight-line extension in a urbanized area. In the first 11 stations, resolved aliphatic (RAli) hydrocarbons presented low values, between 0.07 and 0.96 microg/g dry weight (dw); the unresolved complex mixture (UCM) between 0.42 and 2.72 microg/g dw, and the total aliphatic (TAli) hydrocarbons between 0.55 and 3.07 microg/g dw. In the last station, at the mouth of Chubut river, these values increased to 460, 284, and 741 microg/g dw for RAli, UCM and TAli, respectively. The n-alkanes distribution indices and the compositional parameters suggested a predominantly biogenic origin in eleven stations, and a predominantly anthropogenic origin in the last station, with the highest hydrocarbon values. It is possible to conclude that the stations with low hydrocarbon values and biogenic origin predominance would constitute the baseline of aliphatic hydrocarbons for river sediments at this zone. The station with the highest hydrocarbon concentration and predominantly anthropic origin was related to the presence of Rawson city's port, where its activities (harbor and fishing vessels) generate hydrocarbon wastes unrelated to the river base profile in the study zone. Offshore, but within the river influence, there is an important fishing area of Argentine Red and Patagonian shrimps (Pleoticus muelleri and Artemesia longinaris, respectively). PMID- 15111039 TI - Meiofaunal colonisation on artificial substrates: a tool for biomonitoring the environmental quality on coastal marine systems. AB - The use of artificial substrates for monitoring environmental quality and contaminant impact has been repeatedly suggested, but little information is yet available on the applicability and the sensitivity of this approach. One of the crucial points to be addressed for using artificial substrates, as tools mirroring actual benthic community state, is assessing whether, and how, they reflect in situ conditions characterising natural sediments. In this study we investigated short-term dynamics of meiofaunal colonisation on artificial substrates (bottle brushes) deployed in impacted and "pristine" (control) sites. Meiofaunal colonisation reached a threshold level after 4-6 days in both impacted and control sites but with significantly higher densities in artificial substrates deployed in the control site. Meiofaunal parameters in artificial substrates reflected those of meiofaunal assemblages inhabiting natural sediments where artificial substrates were deployed. Colonised artificial substrates were then transplanted from impacted to control sites. This caused a significant increase of meiofaunal abundance, which after 5 days reached values indistinguishable from the non-impacted control. Given the rapid colonisation time, meiofaunal sensitivity to changing environmental conditions, it can be concluded that artificial substrates can represent an useful tool to be further developed for routine and low-cost monitoring studies aiming at integrating biological indicators of environmental quality. PMID- 15111040 TI - Accumulation and distribution of petroleum hydrocarbons found in mussels (Mytilus galloprovincialis) in the canals of Venice, Italy. AB - Petroleum contamination was assessed in indigenous and transplanted mussels for three types of environments within the Venice Lagoon and its associated interior canals. Indigenous and transplanted mussels were least impacted by petroleum contamination at open-water stations, but more affected in partially-enclosed areas indicating that physical processes of tidal or wind activities can remove organic contaminants such as petroleum from these areas more effectively. Limited tidal flushing of the interior canals resulted in contaminated locations unable to support indigenous mussels. Clean mussels transplanted to these highly impacted sites accumulated the highest levels of petroleum hydrocarbons. The distribution of polycyclic aromatic hydrocarbons (PAHs) indicated mainly combustion-related activities from sources such as the nearby industrial zones and boat traffic, with some evidence of fresher petroleum inputs. Total hydrocarbons found in all mussels could be related to concentrations found in the surrounding sediments as reported in an earlier study. PMID- 15111041 TI - Screening of surfactants for harmful algal blooms mitigation. AB - Screening experiments were conducted in order to find promising synthetic surfactants for harmful algal blooms (HABs) mitigation. The chemically synthesized surfactant cocamidopropyl betaine (CAPB) showed characteristics of relatively high inhibition efficiency, high biodegradability and low cost. The motility inhibition ratios of 10 mg/L CAPB on Cochlodinium polykrikoides and Alexandrium tamarense were about 60% after 5 min. The biodegradation test indicated that the half-life of CAPB in seawater was shorter than one day and 90% was biodegraded after five days under the initial concentration of 100 mg/L at 25 degrees C. Further cell lysis experiments revealed the selective lysis effect of CAPB on different HAB organisms. More than 90% of C. polykrikoides lysed at the concentration of 10 mg/L CAPB after 24 h and at 15 mg/L CAPB after 4 h, whereas the lysis effect of CAPB on A. tamarense was slight, no more than 10% after 2 h interaction with 50 mg/L CAPB. This research provided preliminary data for CAPB as a candidate in harmful algal blooms mitigation and pointed out unresolved problems for its practical application in the meantime. PMID- 15111042 TI - Ecological risk assessment using trace elements from surface sediments of Izmit Bay (Northeastern Marmara Sea) Turkey. AB - The aim of this study was to study ecological risk assessment of trace elements (As, Cd, Cr, Cu, Ni, Pb and Zn) in Izmit Bay surface sediments in the <63 microm fraction. Sediment samples were collected from 8 sites along the north coastline of the bay. Samples were subjected to a total digestion technique and analyzed for major (organic carbon) and minor (As, Cd, Cr, Cu, Ni, Pb and Zn) elements. Sediments heavily contaminated are evaluated by the Sediment Quality Guidelines (SQG) of US EPA. The degree of contamination (Cd) is estimated as very high at each site. Two sets of SQGs, effect range-low/effect range median values and TEL/probable effect level (PEL) values were used in our study. Sediments from the each site are judged toxic when two or more of the PEL values exceed EPA guidelines. Our results are in agreement with previously reported sediment toxicity test results. PMID- 15111043 TI - Deterioration Index (DI): a suggested criterion for assessing the health of coral communities. AB - The extensive deterioration of coral reefs worldwide highlights the importance of creating efficient monitoring methods to best assess their state of health. At present, several suggested parameters serve such indicators. None of these, however, is well accepted as reliably representing reef community health. In the present study we examine a new approach based on the ratio between mortality and recruitment rates of branching corals, which we term 'Deterioration Index' (DI). It aims at providing a quantitative indication of the state of health of reef building coral communities. The method was developed and tested on 16 coral communities on artificially laid rocks along the coast of Eilat, Red Sea (Gulf of Aqaba). In contrast to frequently used indices (i.e. mortality rate, abundance and species richness), which did not demonstrate a consistent result in comparing disturbed vs. undisturbed coral communities, the DI revealed significant differences between these communities. Our results suggest that the use of the DI may enable the detection of disturbed coral communities in one instance monitoring, where the other parameters had failed. The DI, therefore, may provide a comparable quantitative assessment of the deterioration process and its intensity in a coral community. We propose the DI approach as an efficient and applicable tool for coral reef monitoring. PMID- 15111044 TI - Evaluation of sampling devices for the determination of polycyclic aromatic hydrocarbons in surface microlayer coastal waters. AB - The sea surface microlayer (SML) may play an important role on the transport and fate of persistent organic pollutants in the marine environment. In order to evaluate the appropriateness of a number of sampling devices for the analysis of 14 parent polycyclic aromatic hydrocarbons (3-5 aromatic rings), marine SML waters were sampled using a glass plate, a rotating drum/roller, a metal screen and a surface slick sampler. The underlying waters were also sampled for the determination of the corresponding enrichment factors (EF = [C](microlayer)/[C](underlying water)). The EFs were phase dependent, ranging from 1 to 3 for the dissolved phase and between 4 and 7 for the particulate phase. In order to better assess the performance of the different sampling methods, in terms of phase partitioning, the truly dissolved and colloidal phases were also estimated. Generally, no significant differences were found for the enrichment factors provided by the different methods, due to the observed large variability in concentrations that can be attributed to small-scale coastal processes. However, the metal screen is recommended as the most efficient sampling method for the study of PAHs taking into account the amount of water collected versus time. PMID- 15111045 TI - Chemical dispersant effectiveness testing: influence of droplet coalescence. AB - Thermodynamic and kinetic investigations were performed to determine the influence of coalescence of chemically dispersed crude oil droplets in saline waters. For the range of pH (4-10) and salinity (10 per thousand, 30 per thousand, 50 per thousand ) values studied, zeta-potential values ranged from -3 to -10 mV. As the interaction potential values calculated using Derjaguin-Landau Verway-Overbeek (DLVO) theory were negative, the electrostatic barrier did not produce significant resistance to droplet coalescence. Coalescence kinetics of premixed crude oil and chemical dispersant were determined within a range of mean shear rates (Gm = 5, 10, 15, 20 s(-1)) and salinity (10 per thousand, 30 per thousand ) values. Coalescence reaction rates were modeled using Smoluchowski reaction kinetics. Measured collision efficiency values (alpha = 0.25) suggest insignificant resistance to coalescence in shear systems. Experimentally determined dispersant efficiencies (alpha = 0.35) were 10-50% lower than that predicted using a non-interacting droplet model (alpha = 0.0). Unlike other protocols in which the crude oil and dispersant are not premixed, salinity effects were not significant in this protocol. This approach allowed the effects of dispersant-oil contact efficiency eta(contact) to be separated from those of water column transport efficiency (eta(transport)) and coalescence efficiency (eta(coalescence)). PMID- 15111047 TI - Tolerance and accumulation of hexavalent chromium by two seaweed associated aspergilli. AB - Marine seaweed (Eucheuma sp.) associated strains of Aspergillus flavus and Aspergillus niger were tested for their Cr(VI) tolerance. Both the isolates showed luxuriant growth in different concentrations of Cr(VI), i.e., 25, 50 and 100 ppm. There was no marked variation in the dry weight of control and test isolates, which indicated that both the isolates can tolerate a wide range of hexavalent chromium and their application for bioremediation purpose can be envisaged as XRF data revealed both the isolates accumulated more than 25% of the chromium supplied. A. flavus invariably exhibited higher accumulation potential. PMID- 15111046 TI - Desertification of Mediterranean rocky reefs caused by date-mussel, Lithophaga lithophaga (Mollusca: Bivalvia), fishery: effects on adult and juvenile abundance of a temperate fish. AB - Macroalgal beds in temperate rocky habitats provide shelter and food for many adult reef fishes and their juvenile stages. In the Mediterranean Sea, the fishery of the endolithic date-mussel Lithophaga lithophaga (which involves dismantling of rocky substrates inhabited by these bivalves) may cause formation of barrens in shallow rocky reefs. Preliminary data collected in SW Apulia (SE Italy) show that rocky reefs impacted by this destructive fishery display different distribution patterns of adult Coris julis (a common labrid fish in the Mediterranean basin), and lower abundance of juveniles. The ecological implications of date-mussel fishery for dynamics of fish populations and rocky reef ecosystem functioning (e.g., nursery role) are discussed. PMID- 15111048 TI - Trace elements and halogenated organic compounds in Canadian Arctic seabirds. PMID- 15111049 TI - Baseline study of perfluorochemicals in harbour porpoises (Phocoena phocoena) from Northern Europe. PMID- 15111050 TI - Impact of the deposition of jarosite on deepwater meiobenthos in southeastern Australia. PMID- 15111051 TI - Polychlorinated dibenzo-p-dioxins, dibenzofurans and dioxin-like biphenyls in biota from Sri Lankan coast. PMID- 15111052 TI - The N-terminal propeptide of lung surfactant protein C is necessary for biosynthesis and prevents unfolding of a metastable alpha-helix. AB - The lung surfactant-associated protein C (SP-C) consists mainly of a polyvaline alpha-helix, which is stable in a lipid membrane. However, in agreement with the predicted beta-strand conformation of a polyvaline segment, helical SP-C unfolds and transforms into beta-sheet aggregates and amyloid fibrils within a few days in aqueous organic solvents. SP-C fibril formation and aggregation have been associated with lung disease. Here, we show that in a recently isolated biosynthetic precursor of SP-C (SP-Ci), a 12 residue N-terminal propeptide locks the metastable polyvaline part in a helical conformation. The SP-Ci helix does not aggregate or unfold during several weeks of incubation, as judged by hydrogen/deuterium exchange and mass spectrometry. Hydrogen/deuterium exchange experiments further indicate that the propeptide reduces exchange in parts corresponding to mature SP-C. Finally, in an acidic environment, SP-Ci unfolds and aggregates into amyloid fibrils like SP-C. These data suggest a direct role of the N-terminal propeptide in SP-C biosynthesis. PMID- 15111053 TI - ATP-dependent transcriptional activation by bacterial PspF AAA+protein. AB - Transcription activation by bacterial sigma(54)-dependent enhancer-binding proteins (EBPs) requires their tri-nucleotide hydrolysis to restructure the sigma(54) RNA polymerase (RNAP). EBPs share sequence similarity with guanine nucleotide binding-proteins and ATPases associated with various cellular activities (AAA) proteins, especially in the mononucleotide binding P-loop fold. Using the phage shock protein F (PspF) EBP, we identify P-loop residues responsible for nucleotide binding and hydrolysis, consistent with their roles in other P-loop NTPases. We show the refined low-resolution structure of an EBP, PspF, revealing a hexameric ring organisation characteristic of AAA proteins. Functioning of EBPs involves ATP binding, higher oligomer formation and ATP hydrolysis coupled to the restructuring of the RNAP. This is thought to be a highly coordinated multi-step process, but the nucleotide-driven mechanism of oligomerisation and ATP hydrolysis is little understood. Our kinetic and structural data strongly suggest that three PspF dimers assemble to form a hexamer upon nucleotide binding. During the ATP hydrolysis cycle, both ATP and ADP are bound to oligomeric PspF, in line with a sequential hydrolysis cycle. We identify a putative R-finger, and show its involvement in ATP hydrolysis. Substitution of this arginine residue results in nucleotide-independent formation of hexameric rings, structurally linking the putative R-finger and, by inference, a specific nucleotide interaction to the control of PspF oligomerisation. PMID- 15111054 TI - Distinct DNA elements contribute to Rap1p affinity for its binding sites. AB - The essential Saccharomyces cerevisiae regulatory protein Rap1 contains two tandem Myb-like DNA binding sub-domains that interact with two defined DNA "hemisites", separated by a trinucleotide linker sequence. We have mapped the thermodynamically defined DNA-binding site of Rap1 by a primer extension method coupled with electrophoretic separation of bound and unbound DNAs. Relative to published consensus sequences, we detect binding interactions that extend 3 bp beyond the 5'-end of the putative DNA-binding site. This new site of interaction is located where the DNA minor groove faces the protein, and may account for the major DNA bending induced by Rap1p that previous studies have mapped to a site immediately upstream of the consensus binding site. In addition, we show that a minimal DNA-binding site made of one single consensus hemisite, preceded or followed by a spacer trinucleotide that interacts with the unstructured protein linker between the two Rap1p DNA binding domains, is able to bind the protein, although at lower affinity. These findings may explain the observed in vivo binding properties of Rap1p at many promoters that lack canonical binding sites. PMID- 15111055 TI - Analysis of human tyrosyl-DNA phosphodiesterase I catalytic residues. AB - Tyrosyl-DNA phosphodiesterase I (Tdp1) is involved in the repair of DNA lesions created by topoisomerase I in vivo. Tdp1 is a member of the phospholipase D (PLD) superfamily of enzymes and hydrolyzes 3'-phosphotyrosyl bonds to generate 3' phosphate DNA and free tyrosine in vitro. Here, we use synthetic 3'-(4 nitro)phenyl, 3'-(4-methyl)phenyl, and 3'-tyrosine phosphate oligonucleotides to study human Tdp1. Kinetic analysis of human Tdp1 (hTdp1) shows that the enzyme has nanomolar affinity for all three substrates and the overall in vitro reaction is diffusion-limited. Analysis of active-site mutants using these modified substrates demonstrates that hTdp1 uses an acid/base catalytic mechanism. The results show that histidine 493 serves as the general acid during the initial transesterification, in agreement with hypotheses based on previous crystal structure models. The results also argue that lysine 495 and asparagine 516 participate in the general acid reaction, and the analysis of crystal structures suggests that these residues may function in a proton relay. Together with previous crystal structure data, the new functional data provide a mechanistic understanding of the conserved histidine, lysine and asparagine residues found among all PLD family members. PMID- 15111056 TI - Selection and characterization of small random transmembrane proteins that bind and activate the platelet-derived growth factor beta receptor. AB - Growth factor receptors are typically activated by the binding of soluble ligands to the extracellular domain of the receptor, but certain viral transmembrane proteins can induce growth factor receptor activation by binding to the receptor transmembrane domain. For example, homodimers of the transmembrane 44-amino acid bovine papillomavirus E5 protein bind the transmembrane region of the PDGF beta receptor tyrosine kinase, causing receptor dimerization, phosphorylation, and cell transformation. To determine whether it is possible to select novel biologically active transmembrane proteins that can activate growth factor receptors, we constructed and identified small proteins with random hydrophobic transmembrane domains that can bind and activate the PDGF beta receptor. Remarkably, cell transformation was induced by approximately 10% of the clones in a library in which 15 transmembrane amino acid residues of the E5 protein were replaced with random hydrophobic sequences. The transformation-competent transmembrane proteins formed dimers and stably bound and activated the PDGF beta receptor. Genetic studies demonstrated that the biological activity of the transformation-competent proteins depended on specific interactions with the transmembrane domain of the PDGF beta receptor. A consensus sequence distinct from the wild-type E5 sequence was identified that restored transforming activity to a non-transforming poly-leucine transmembrane sequence, indicating that divergent transmembrane sequence motifs can activate the PDGF beta receptor. Molecular modeling suggested that diverse transforming sequences shared similar protein structure, including the same homodimer interface as the wild-type E5 protein. These experiments have identified novel proteins with transmembrane sequences distinct from the E5 protein that can activate the PDGF beta receptor and transform cells. More generally, this approach may allow the creation and identification of small proteins that modulate the activity of a variety of cellular transmembrane proteins. PMID- 15111057 TI - Solution structure determination of monomeric human IgA2 by X-ray and neutron scattering, analytical ultracentrifugation and constrained modelling: a comparison with monomeric human IgA1. AB - Immunoglobulin A (IgA), the most abundant human immunoglobulin, mediates immune protection at mucosal surfaces as well as in plasma. It exists as two subclasses IgA1 and IgA2, and IgA2 is found in at least two allotypic forms, IgA2m(1) or IgA2m(2). Compared to IgA1, IgA2 has a much shorter hinge region, which joins the two Fab and one Fc fragments. In order to assess its solution structure, monomeric recombinant IgA2m(1) was studied by X-ray and neutron scattering. Its Guinier X-ray radius of gyration R(G) is 5.18 nm and its neutron R(G) is 5.03 nm, both of which are significantly smaller than those for monomeric IgA1 at 6.1-6.2 nm. The distance distribution function P(r)for IgA2m(1) showed a broad peak with a subpeak and gave a maximum dimension of 17 nm, in contrast to the P(r) curve for IgA1, which showed two distinct peaks and a maximum dimension of 21 nm. The sedimentation coefficients of IgA1 and IgA2m(1) were 6.2S and 6.4S, respectively. These data show that the solution structure of IgA2m(1) is significantly more compact than IgA1. The complete monomeric IgA2m(1) structure was modelled using molecular dynamics to generate random IgA2 hinge structures, to which homology models for the Fab and Fc fragments were connected to generate 10,000 full models. A total of 104 compact best-fit IgA2m(1) models gave good curve fits. These best-fit models were modified by linking the two Fab light chains with a disulphide bridge that is found in IgA2m(1), and subjecting these to energy refinement to optimise this linkage. The averaged solution structure of the arrangement of the Fab and Fc fragments in IgA2m(1) was found to be predominantly T-shaped and flexible, but also included Y-shaped structures. The IgA2 models show full steric access to the two FcalphaRI-binding sites at the Calpha2-Calpha3 interdomain region in the Fc fragment. Since previous scattering modelling had shown that IgA1 also possessed a flexible T-shaped solution structure, such a T shape may be common to both IgA1 and IgA2. The final models suggest that the combination of the more compact IgA2m(1) and the more extended IgA1 structures will enable human IgA to access a broader range of antigens than either acting alone. The hinges of both IgA subclasses appear to show reduced flexibility when compared to their equivalents in IgG, and this may be important for maintaining an extended IgA structure. PMID- 15111058 TI - Monitoring the process of HypF fibrillization and liposome permeabilization by protofibrils. AB - Much information has appeared in the last few years on the low resolution structure of amyloid fibrils and on their non-fibrillar precursors formed by a number of proteins and peptides associated with amyloid diseases. The fine structure and the dynamics of the process leading misfolded molecules to aggregate into amyloid assemblies are far from being fully understood. Evidence has been provided in the last five years that protein aggregation and aggregate toxicity are rather generic processes, possibly affecting all polypeptide chains under suitable experimental conditions. This evidence extends the number of model proteins one can investigate to assess the molecular bases and general features of protein aggregation and aggregate toxicity. We have used tapping mode atomic force microscopy to investigate the morphological features of the pre-fibrillar aggregates and of the mature fibrils produced by the aggregation of the hydrogenase maturation factor HypF N-terminal domain (HypF-N), a protein not associated to any amyloid disease. We have also studied the aggregate-induced permeabilization of liposomes by fluorescence techniques. Our results show that HypF-N aggregation follows a hierarchical path whereby initial globules assemble into crescents; these generate large rings, which evolve into ribbons, further organizing into differently supercoiled fibrils. The early pre-fibrillar aggregates were shown to be able to permeabilize synthetic phospholipid membranes, thus showing that this disease-unrelated protein displays the same amyloidogenic behaviour found for the aggregates of most pathological proteins and peptides. These data complement previously reported findings, and support the idea that protein aggregation, aggregate structure and toxicity are generic properties of polypeptide chains. PMID- 15111059 TI - Crystallographic structure and biochemical analysis of the Thermus thermophilus osmotically inducible protein C. AB - The X-ray crystallographic structure of osmotically inducible Protein C from the thermophilic bacterium, Thermus thermophilus HB8, was solved to 1.6A using the multiple wavelength anomalous dispersion method and a selenomethionine incorporated protein (Se-MAD). The crystal space group was P1 with cell dimensions of a=37.58 A, b=40.95 A, c=48.14 A, alpha=76.9 degrees, beta=74.0 degrees and gamma=64.1 degrees. The two tightly interacting monomers in the asymmetric unit are related by a non-crystallographic 2-fold. The dimer structure is defined primarily by two very long anti-parallel, over-lapping alpha-helices at the core, with a further six-stranded anti-parallel beta-sheet on the outside of the structure. With respect to the beta-sheets, both A and B monomers contribute three strands each resulting in an intertwining of the structure. The active site consists of two cysteine residues from one monomer and an arginine and glutamic acid from the other. Enzymatic assays have revealed that T.thermophilus OsmC has a hydroperoxide peroxidase activity. PMID- 15111060 TI - A kinetic analysis of the nucleotide-induced allosteric transitions in a single ring mutant of GroEL. AB - The function of GroE requires a complex system of allosteric communication driven by protein-nucleotide interactions. These rearrangements couple the binding and hydrolysis of ATP to an overall reaction cycle in which substrate proteins are bound, encapsulated and released. Positive cooperativity with respect to ATP binding occurs within one heptameric ring of GroEL, while negative cooperativity between the two rings generates an inherent asymmetry between the two rings. A previously engineered mutant of GroEL in which the ring-ring contacts are broken gives rise to a single-ring version of the wild-type chaperonin (SR1). We have studied the kinetics of the nucleotide-induced conformational changes in a single tryptophan variant of SR1 (Y485W-SR1) and compared the resulting data with those we reported previously for the double-ring, single-tryptophan variant of wild type GroEL (Y485W-GroEL). Remarkably, the parting of the rings does not have a major effect on the conformational changes occurring within the heptameric ring and a kinetic model is presented to describe the sequence of structural rearrangements that occur upon ATP binding to the SR1 molecule. The observation that both the ATP-induced and ADP-induced conformational rearrangements occur more rapidly in the SR1 than they do in wild-type GroEL, indicates that intra ring conformational changes in the double-ring structure must overcome conformational constraints provided by the presence of the second ring. Lastly, the data presented here imply a role for inter-ring allostery in controlling the dissociation-association behaviour of the GroES co-protein in the overall reaction cycle. PMID- 15111061 TI - Kinetic analysis of ATP-dependent inter-ring communication in GroEL. AB - Different concentrations of ATP were mixed rapidly with single-ring or double ring forms of GroEL containing the Phe44-->Trp mutation and the time-resolved changes in fluorescence emission, upon excitation at 295 nm, were followed. Two kinetic phases that were previously found for double-ring GroEL are also observed in the case of the single-ring version: (i) a fast phase with a relatively large amplitude that is associated with the T-->R allosteric transition; (ii) and a slow phase with a smaller amplitude that is associated with ATP hydrolysis. In the case of weak intra-ring positive cooperativity, the rate constant corresponding to the T-->R allosteric switch of single-ring GroEL displays a bi sigmoidal dependence on ATP concentration that may reflect parallel pathways of the allosteric transition. The slow phase is absent when double-ring or single ring forms of GroEL are mixed with ADP or ATP without K(+), and it has a rate constant that is independent of ATP concentration. A third fast phase that is still unassigned is observed for both single-ring and double-ring GroEL when a large amount of data is collected. Finally, a fourth phase is observed in the case of double-ring GroEL that is found to be absent in the case of single-ring GroEL. This phase is here assigned to inter-ring communication by (i) determining its dependence on ATP concentration and (ii) based on its absence from single ring GroEL and the Arg13-->Gly, Ala126-->Val GroEL mutant, which is defective in inter-ring negative cooperativity. The value of the rate constant corresponding to this phase is found to increase with increasing intra-ring positive cooperativity, with respect to ATP. This is the first report of the rate of ATP mediated inter-ring communication in GroEL, in the presence of ATP alone, which is crucial for the cycling of this molecular machine between different functional states. PMID- 15111062 TI - Thermostability gradient in the collagen triple helix reveals its multi-domain structure. AB - A triple-helical conformation and stability at physiological temperature are critical for the mechanical and biological functions of the fibril-forming collagens. Here, we characterized the role of consecutive domains of collagen II in stabilizing the triple helix. Analysis of melting temperatures of genetically engineered collagen-like proteins consisting of tandem repeats of the D1, D2, D3 or D4 collagen II periods revealed the presence of a gradient of thermostability along the collagen molecule with thermolabile N-terminal domains and thermostable C-terminal domains. These results imply a multi-domain character of the collagen triple helix. Assays of thermostabilities of the Arg75Cys and Arg789Cys collagen II mutants suggest that, in contrast to the thermostable domains, the thermolabile domains are able to accommodate amino acid substitutions without altering the thermostability of the entire collagen molecule. PMID- 15111063 TI - The 0.83 A resolution crystal structure of alpha-lytic protease reveals the detailed structure of the active site and identifies a source of conformational strain. AB - The crystal structure of the extracellular bacterial serine protease alpha-lytic protease (alphaLP) has been solved at 0.83 A resolution at pH 8. This ultra-high resolution structure allows accurate analysis of structural elements not possible with previous structures. Hydrogen atoms are visible, and confirm active-site hydrogen-bonding interactions expected for the apo enzyme. In particular, His57 N(delta1) participates in a normal hydrogen bond with Asp102 in the catalytic triad, with a hydrogen atom visible 0.83(+/-0.06)A from the His N(delta1). The catalytic Ser195 occupies two conformations, one corresponding to a population of His57 that is doubly protonated, the other to the singly protonated His57. Based on the occupancy of these conformations, the pKa of His57 is calculated to be approximately 8.8 when a sulfate ion occupies the active site. This 0.83 A structure has allowed critical analysis of geometric distortions within the structure. Interestingly, Phe228 is significantly distorted from planarity. The distortion of Phe228, buried in the core of the C-terminal domain, occurs at an estimated energetic cost of 4.1 kcal/mol. The conformational space for Phe228 is severely limited by the presence of Trp199, which prevents Phe228 from adopting the rotamer observed in many other chymotrypsin family members. In alphaLP, the only allowed rotamer leads to the deformation of Phe228 due to steric interactions with Thr181. We hypothesize that tight packing of co-evolved residues in this region, and the subsequent deformation of Phe228, contributes to the high cooperativity and large energetic barriers for folding and unfolding of alphaLP. The kinetic stability imparted by the large, cooperative unfolding barrier plays a critical role in extending the lifetime of the protease in its harsh environment. PMID- 15111064 TI - Preformed structural elements feature in partner recognition by intrinsically unstructured proteins. AB - Intrinsically unstructured proteins (IUPs) are devoid of extensive structural order but often display signs of local and limited residual structure. To explain their effective functioning, we reasoned that such residual structure can be crucial in their interactions with their structured partner(s) in a way that preformed structural elements presage their final conformational state. To check this assumption, a database of 24 IUPs with known 3D structures in the bound state has been assembled and the distribution of secondary structure elements and backbone torsion angles have been analysed. The high proportion of residues in coil conformation and with phi, psi angles in the disallowed regions of the Ramachandran map compared to the reference set of globular proteins shows that IUPs are not fully ordered even in their bound form. To probe the effect of partner proteins on IUP folding, inherent conformational preferences of IUP sequences have been assessed by secondary structure predictions using the GOR, ALB and PROF algorithms. The accuracy of predicting secondary structure elements of IUPs is similar to that of their partner proteins and is significantly higher than the corresponding values for random sequences. We propose that strong conformational preferences mark regions in IUPs (mostly helices), which correspond to their final structural state, while regions with weak conformational preferences represent flexible linkers between them. In our interpretation, preformed elements could serve as initial contact points, the binding of which facilitates the reeling of the flexible regions onto the template. This finding implies that IUPs draw a functional advantage from preformed structural elements, as they enable their facile, kinetically and energetically less demanding, interaction with their physiological partner. PMID- 15111065 TI - A combined transmembrane topology and signal peptide prediction method. AB - An inherent problem in transmembrane protein topology prediction and signal peptide prediction is the high similarity between the hydrophobic regions of a transmembrane helix and that of a signal peptide, leading to cross-reaction between the two types of predictions. To improve predictions further, it is therefore important to make a predictor that aims to discriminate between the two classes. In addition, topology information can be gained when successfully predicting a signal peptide leading a transmembrane protein since it dictates that the N terminus of the mature protein must be on the non-cytoplasmic side of the membrane. Here, we present Phobius, a combined transmembrane protein topology and signal peptide predictor. The predictor is based on a hidden Markov model (HMM) that models the different sequence regions of a signal peptide and the different regions of a transmembrane protein in a series of interconnected states. Training was done on a newly assembled and curated dataset. Compared to TMHMM and SignalP, errors coming from cross-prediction between transmembrane segments and signal peptides were reduced substantially by Phobius. False classifications of signal peptides were reduced from 26.1% to 3.9% and false classifications of transmembrane helices were reduced from 19.0% to 7.7%. Phobius was applied to the proteomes of Homo sapiens and Escherichia coli. Here we also noted a drastic reduction of false classifications compared to TMHMM/SignalP, suggesting that Phobius is well suited for whole-genome annotation of signal peptides and transmembrane regions. The method is available at as well as at PMID- 15111066 TI - Biomechanics of microscopic appendages: functional shifts caused by changes in speed. AB - Many diverse animals use arrays of hair-like structures to perform important jobs such as feeding, gas exchange, smelling, and swimming. Since these functions involve hair interactions with the surrounding water or air, analysis of the fluid dynamics of diverse hair-bearing appendages reveals how the morphology of an array of hairs affects it performance. Mathematical and physical models of flow between cylinders have shown that arrays of large, rapidly moving cylinders are leaky sieves, whereas little fluid moves through a row of small, slow rods. The purpose of the present study was to test this prediction for realistic appendage morphologies and to elucidate whether the design of a hairy leg can affect the range of speeds in which this transition in function occurs. We studied flow through hairy food-capturing appendages (second maxillae) of calanoid copepods, abundant planktonic crustaceans whose feeding on unicellular algae forms an important link in many marine food webs. Using dynamically scaled physical models, we found that hairy appendages undergo a transition between paddle- and sieve-like function at a critical range of sizes and speeds. The coarser the mesh of hairs on second maxillae, the smaller the size and speed at which this functional shift occurs. Thus, a simple increase in size (ontogenetic or evolutionary) or speed can generate a novel function (a paddle can become a filter), but the morphology of a hairy appendage determines the size and speed range at which leakiness to fluid flow can be affected by behavior or growth. PMID- 15111067 TI - Pattern of anterior cruciate ligament force in normal walking. AB - The goal of this study was to calculate and explain the pattern of anterior cruciate ligament (ACL) loading during normal level walking. Knee-ligament forces were obtained by a two-step procedure. First, a three-dimensional (3D) model of the whole body was used together with dynamic optimization theory to calculate body-segmental motions, ground reaction forces, and leg-muscle forces for one cycle of gait. Joint angles, ground reaction forces, and muscle forces obtained from the gait simulation were then input into a musculoskeletal model of the lower limb that incorporated a 3D model of the knee. The relative positions of the femur, tibia, and patella and the forces induced in the knee ligaments were found by solving a static equilibrium problem at each instant during the simulated gait cycle. The model simulation predicted that the ACL bears load throughout stance. Peak force in the ACL (303 N) occurred at the beginning of single-leg stance (i.e., contralateral toe off). The pattern of ACL force was explained by the shear forces acting at the knee. The balance of muscle forces, ground reaction forces, and joint contact forces applied to the leg determined the magnitude and direction of the total shear force acting at the knee. The ACL was loaded whenever the total shear force pointed anteriorly. In early stance, the anterior shear force from the patellar tendon dominated the total shear force applied to the leg, and so maximum force was transmitted to the ACL at this time. ACL force was small in late stance because the anterior shear forces supplied by the patellar tendon, gastrocnemius, and tibiofemoral contact were nearly balanced by the posterior component of the ground reaction. PMID- 15111068 TI - Residual stress and strain in the lamellar unit of the porcine aorta: experiment and analysis. AB - The opened-up configuration of the artery wall has long been assumed to be stress free. This is questionable in a microscopic level. The aortic media is made of concentric layers whose unit is called a lamellar unit, a pair of elastic lamina (EL) and a smooth muscle-rich layer (SML). Recently, we found that the EL was about 2.5 times stiffer than the SML. If the circumferential stress in the in vivo condition is the same between the two layers, residual stress of each layer should be different because the stress-strain relationships differ. Such residual stress is not released fully by radial cutting, but is released in the area close to the cut surface, causing hills and valleys on the surface due to residual stress. To check this hypothesis, we have developed a scanning micro indentation tester, a scaled-up version of the atomic force microscope, and measured the topography and the stiffness distribution of the cut surface. The surface of the section of porcine thoracic aortas shows hill and valley pattern corresponding with their histology. The hills were more than three times stiffer than the valleys, indicating that the hills are the ELs and the valleys the SMLs, and the ELs are compressed and the SMLs stretched in the lamellar unit. A finite element analysis showed that the residual stress in the EL and the SML is much higher than those estimated in the unloaded ring-like segments. Fairly large stress may still reside in the opened-up aortic wall. PMID- 15111069 TI - Muscle mechanical work requirements during normal walking: the energetic cost of raising the body's center-of-mass is significant. AB - Inverted pendulum models of walking predict that little muscle work is required for the exchange of body potential and kinetic energy in single-limb support. External power during walking (product of the measured ground reaction force and body center-of-mass (COM) velocity) is often analyzed to deduce net work output or mechanical energetic cost by muscles. Based on external power analyses and inverted pendulum theory, it has been suggested that a primary mechanical energetic cost may be associated with the mechanical work required to redirect the COM motion at the step-to-step transition. However, these models do not capture the multi-muscle, multi-segmental properties of walking, co-excitation of muscles to coordinate segmental energetic flow, and simultaneous production of positive and negative muscle work. In this study, a muscle-actuated forward dynamic simulation of walking was used to assess whether: (1). potential and kinetic energy of the body are exchanged with little muscle work; (2). external mechanical power can estimate the mechanical energetic cost for muscles; and (3.) the net work output and the mechanical energetic cost for muscles occurs mostly in double support. We found that the net work output by muscles cannot be estimated from external power and was the highest when the COM moved upward in early single-limb support even though kinetic and potential energy were exchanged, and muscle mechanical (and most likely metabolic) energetic cost is dominated not only by the need to redirect the COM in double support but also by the need to raise the COM in single support. PMID- 15111070 TI - Mechanical and metabolic requirements for active lateral stabilization in human walking. AB - Walking appears to be passively unstable in the lateral direction, requiring active feedback control for stability. The central nervous system may control stability by adjusting medio-lateral foot placement, but potentially with a metabolic cost. This cost increases with narrow steps and may affect the preferred step width. We hypothesized that external stabilization of the body would reduce the active control needed, thereby decreasing metabolic cost and preferred step width. To test these hypotheses, we provided external lateral stabilization, using springs pulling bilaterally from the waist, to human subjects walking on a force treadmill at 1.25 m/s. Ten subjects walked, with and without stabilization, at a prescribed step width of zero and also at their preferred step width. We measured metabolic cost using indirect calorimetry, and step width from force treadmill data. We found that at the prescribed zero step width, external stabilization resulted in a 33% decrease in step width variability (root-mean-square) and a 9.2% decrease in metabolic cost. In the preferred step width conditions, external stabilization caused subjects to prefer a 47% narrower step width, with a 32% decrease in step width variability and a 5.7% decrease in metabolic cost. These results suggest that (a). human walking requires active lateral stabilization, (b). body lateral motion is partially stabilized via medio-lateral foot placement, (c). active stabilization exacts a modest metabolic cost, and (d). humans avoid narrow step widths because they are less stable. PMID- 15111071 TI - Age-related reduction in sagittal plane center of mass motion during obstacle crossing. AB - Accidental falls are a leading cause of injury and death in the growing elderly population. Traumatic falls are frequent, costly, and debilitating. Control of balance during locomotion is critical for safe ambulation, but relatively little is known about the natural effect of aging on dynamic balance control. Samples of healthy young (n = 13) and elderly (n = 13) subjects were compared in the interactive measures of center of mass (COM) and center of pressure (COP) during level walking and obstacle crossing conditions. Obstacle heights were normalized to individual body height (2.5%, 5%, 10%, and 15%). Temporal-distance (T-D) variables of gait were also compared. Statistical analyses were conducted using a two-way ANOVA for subject group and obstacle height. T-D parameters were not significantly different between groups; nor were frontal plane COM and COP parameters. Significant age differences did exist for antero-posterior (A/P) motion of the COM (decreased motion in the elderly), and its relationship with the COP (reduced separation between the two variables in the elderly). Anterior COM velocities were also significantly lower in the elderly group. The results confirm the ability of healthy elderly adults to maintain dynamic balance control in the frontal plane during locomotion. Reduced A/P distances between the COM and COP indicate a conservative reduction of the mechanical load on joints of the supporting limb. This conservative strategy may be related to a reduction in muscle strength as it occurs in the natural aging process. PMID- 15111072 TI - Frequency-dependent changes in neuromuscular responses to cyclic lumbar flexion. AB - Repetitive lifting in the workplace has been identified to be a cause of low back disorders. Epidemiologic data further supports an hypothesis that higher repetition rate (i.e. frequency) is an added risk factor. The objective of this study was to provide experimental data testing the above hypothesis. An in vivo feline model was subjected to 20-min of cyclic lumbar loading at frequencies of 0.1 Hz and 0.5 Hz while monitoring the EMG from the L-3/4-L-5/6 multifidus muscles and the creep at the L-4/5 level. Seven hours of rest were allowed after the cyclic flexion/extension was terminated. During this rest period, a single test cycle was performed every hour to assess recovery of EMG and lumbar creep. The results demonstrate that cyclic lumbar flexion elicits a transient neuromuscular disorder consisting of EMG spasms during the cyclic loading and initial and delayed muscular hyperexcitabilities during the rest period. Cyclic loading at 0.5 Hz resulted in significant (p<0.05) increase in the hyperexcitability magnitude and duration during the recovery period. It was concluded that repetitive lumbar loading at fast rates is indeed a risk factor as it induces larger creep in the lumbar viscoelastic tissues which in turn intensify the resulting neuromuscular disorder. PMID- 15111073 TI - Frequency content asymmetry of the isokinetic curve between ACL deficient and healthy knee. AB - The torque-time curve patterns of concentric isokinetic knee extension in anterior cruciate ligament (ACL) deficient patients usually present mid-range irregularities associated with the level of anterior tibial translation. The purpose of this study was to compare the smoothness in isokinetic torque production between the ACL deficient and the healthy knee. Thirty ACL deficient soccer players performed bilaterally five trials of maximum concentric knee extension-flexion at 60 degrees /s on a Biodex dynamometer. The three middle trials (a total of six curves) were retained and submitted to further data processing. Maximum frequency values contained within the 90%, 95% and 99% level of the signal power were calculated for each extension and flexion curve. The frequency content of the ACL deficient side proved to be statistically higher compared to the intact side at all levels of the power spectrum. The percentage differences in the frequency content were 18.8%, 10.6% and 40.0% for knee extension, and 49.5%, 24.5% and 16.3% for knee flexion, for the respective power levels. This indicated higher oscillations and, therefore, more unstable mechanical output of the injured knee. An overall biological interpretation of the present results is based on the notion that disturbed motion is generally connected to poor level of joint functionality. PMID- 15111074 TI - Biomechanics of tendon injury and repair. AB - Many clinical and experimental studies have investigated how tendons repair in response to an injury. This body of work has led to a greater understanding of tendon healing mechanisms and subsequently to an improvement in their treatment. In this review paper, characterization of normal and healing tendons is first covered. In addition, the debate between intrinsic and extrinsic healing is examined, and the cellular and extracellular matrix response following a tendon injury is detailed. Next, clinical and experimental injury and repair methods utilizing animal models are discussed. Animal models have been utilized to study the effect of various activity levels, motions, injury methods, and injury locations on tendon injury and repair. Finally, current and future treatment modalities for improving tendon healing, such as tissue engineering, cell therapy, and gene therapy, are reviewed. PMID- 15111075 TI - Differences between collagen morphologies, properties and distribution in diabetic and normal biobreeding and Sprague-Dawley rat sciatic nerves. AB - Both structural and functional differences between normal and diabetic nerve have been observed, in human patients and animal models. We hypothesize that these structural differences are quantifiable, morphologically and mechanically, with the ultimate aim of understanding the contribution of these differences to permanent nerve damage. The outer collagenous epineurial and perineurial tissues of mammalian peripheral nerves mechanically and chemically shield the conducting axons. We have quantified differences in these collagens, using whole-nerve uniaxial testing, and immunohistochemistry of collagens type I, III, and IV in diabetic and normal nerves. We present results of two studies, on normal and diabetic BioBreeding (BB), and normal, diabetic and weight-controlled Sprague Dawley (SD) rats, respectively. Overall, we measured slightly higher uniaxial moduli (e.g. 5.9 MPa vs. 3.5 MPa, BB; 10.7 MPa vs. 10.0 MPa, SD at 40% strain) in whole nerves as well as higher peak stresses (e.g. 0.99 MPa vs. 0.74 MPa, BB; 2.16 MPa vs. 1.94 MPa, SD at 40% strain) in the diabetics of both animal models. We measured increased concentrations of types III and IV collagens in the diabetics of both models and mixed upregulation results were observed in type I protein levels. We detected small differences in mechanical properties at the tissue scale, though we found significant structural and morphometric differences at the fibril scale. These findings suggest that whole-tissue mechanical testing is not a sufficient assay for collagen glycation, and that fibrillar and molecular scale assays are needed to detect the earliest stages of diabetic protein glycation. PMID- 15111076 TI - The effects of walking speed on obstacle crossing in healthy young and healthy older adults. AB - The effects of walking speed and age on the peak external moments generated about the joints of the trailing limb during stance just prior to stepping over an obstacle and on the kinematics of the trailing limb when crossing the obstacle were investigated in 10 healthy young adults (YA) and 10 healthy older adults (OA). The peak hip and knee adduction moments in OA were 21-43% greater than those in YA (p80 FMRFamide-related peptides have been identified in nematodes, only four FMRFamide-related peptides have been characterised from flatworms. The Ascaris suum ovijector/body wall bioassay and the Procerodes littoralis muscle fibre bioassay have proved both reliable and sensitive systems for assessing the functional activities of FMRFamide-related peptides in vitro, and data describing the effects of native FMRFamide-related peptides in these systems are rapidly accumulating. This is the first study to determine the cross-phyla activities of non-native FMRFamide-related peptides in both nematode and flatworm species. In the present study, the effects of 10 arthropod FMRFamide-related peptides (leucomyosuppressin [pQDVDHVFLRFamide], schistoFLRFamide [PDVDHVFLRFamide] and truncated analogues [HVFLRFamide and VFLRFamide], lobster peptide I [TNRNFLRFamide], lobster peptide II [SDRNFLRFamide], manducaFLRFamide II [GNSFLRFamide], manducaFLRFamide III [DPSFLRFamide], calliFMRFamide 4 [KPNQDFMRFamide] and perisulfakinin [EQFDDY(SO(3)H)GHMRFamide]), representing the five subfamilies, were examined on the body wall and ovijector of the parasitic porcine nematode, A. suum and dispersed muscle fibres from the free-living turbellarian, P. littoralis. The muscle activity of the ovijector was found to be modulated significantly by each of the arthropod FMRFamide-related peptides tested; the effects were concentration-dependent, reversible and repeatable. All but one (perisulfakinin) of the 10 arthropod FMRFamide-related peptides examined modulated significantly the activity of A. suum body wall muscle. In addition, all of the arthropod FMRFamide-related peptides examined induced potent concentration-dependent contractions of P. littoralis muscle fibres. These results reveal similarities in the ligand requirement(s) between FMRFamide-related peptide receptors within the Phyla Arthropoda, Nematoda and Platyhelminthes, and indicate significant receptor promiscuity, which highlights the potential of FMRFamide-related peptide receptors as legitimate targets for novel endectocidal agents. PMID- 15111098 TI - On the mechanism of SPP-catalysed intramembrane proteolysis; conformational control of peptide bond hydrolysis in the plane of the membrane. AB - Intramembrane-cleaving proteases are members of a novel type of enzyme that hydrolyse substrate proteins within transmembrane regions. The presently known proteases that catalyse such cleavage reactions are membrane proteins of high hydrophobicity and multiple predicted transmembrane regions. A key feature is the positioning of active site residues in hydrophobic segments implying that the catalytic centre is assembled within the plane of the membrane. Nevertheless, all these proteases appear to utilise catalytic mechanisms similar to classic proteases that expose their active site domains in aqueous compartments. In the present review, we will address the mechanism of intramembrane proteolysis on the example of the signal peptide peptidase, and discuss how enzyme-catalysed hydrolysis of peptide bonds within the plane of a cellular membrane might occur. PMID- 15111099 TI - The archaeal sensory rhodopsin II/transducer complex: a model for transmembrane signal transfer. AB - Archaebacterial photoreceptors mediate phototaxis by regulating cell motility through two-component signalling cascades. Homologs of this sensory pathway occur in all three kingdoms of life, most notably in enteric bacteria in which the chemotaxis has been extensively studied. Recent structural and functional studies on the sensory rhodopsin II/transducer complex mediating the photophobic response of Natronomonas pharaonis have yielded new insights into the mechanisms of signal transfer across the membrane. Electron paramagnetic resonance data and the atomic resolution structure of the receptor molecule in complex with the transmembrane segment of its cognate transducer provided a model for signal transfer from the receptor to the cytoplasmic side of the transducer. This mechanism might also be relevant for eubacterial chemoreceptor signalling. PMID- 15111100 TI - Crystal structure of human monoamine oxidase B, a drug target enzyme monotopically inserted into the mitochondrial outer membrane. AB - Monoamine oxidase B (MAO B) is an outer mitochondrial membrane protein that oxidizes arylalkylamine neurotransmitters and has been a valuable drug target for many neurological disorders. The 1.7 angstrom resolution structure of human MAO B shows the enzyme is dimeric with a C-terminal transmembrane helix protruding from each monomer and anchoring the protein to the membrane. This helix departs perpendicularly from the base of the structure in a different way with respect to other monotopic membrane proteins. Several apolar loops exposed on the protein surface are located in proximity of the C-terminal helix, providing additional membrane-binding interactions. One of these loops (residues 99-112) also functions in opening and closing the MAO B active site cavity, which suggests that the membrane may have a role in controlling substrate binding. PMID- 15111101 TI - Structural basis for ion conduction and gating in ClC chloride channels. AB - Members of the ClC family of voltage-gated chloride channels are found from bacteria to mammals with a considerable degree of conservation in the membrane inserted, pore-forming region. The crystal structures of the ClC channels of Escherichia coli and Salmonella typhimurium provide a structural framework for the entire family. The ClC channels are homodimeric proteins with an overall rhombus-like shape. Each ClC dimer has two pores each contained within a single subunit. The ClC subunit consists of two roughly repeated halves that span the membrane with opposite orientations. This antiparallel architecture defines a chloride selectivity filter within the 15-A neck of a hourglass-shaped pore. Three Cl(-) binding sites within the selectivity filter stabilize ions by interactions with alpha-helix dipoles and by chemical interactions with nitrogen atoms and hydroxyl groups of residues in the protein. The Cl(-) binding site nearest the extracellular solution can be occupied either by a Cl(-) ion or by a glutamate carboxyl group. Mutations of this glutamate residue in Torpedo ray ClC channels alter gating in electrophysiological assays. These findings reveal a form of gating in which the glutamate carboxyl group closes the pore by mimicking a Cl(-) ion. PMID- 15111102 TI - New insights into the structure and oligomeric state of the bacterial multidrug transporter EmrE: an unusual asymmetric homo-dimer. AB - EmrE is a small multidrug transporter that contains 110 amino acid residues that form four transmembrane alpha-helices. The three-dimensional structure of EmrE has been determined from two-dimensional crystals by electron cryo-microscopy. EmrE is an asymmetric homo-dimer with one substrate molecule bound in a chamber accessible laterally from one leaflet of the lipid bilayer. Evidence from substrate binding analyses and analytical ultracentrifugation of detergent solubilised EmrE shows that the minimum functional unit for substrate binding is a dimer. However, it is possible that EmrE exists as a tetramer in vivo and plausible models are suggested based upon analyses of two-dimensional crystals. PMID- 15111103 TI - The role and structure of mitochondrial carriers. AB - Members of the mitochondrial carrier family transport compounds over the inner mitochondrial membrane to link the biochemical pathways in the cytosol with those in the mitochondrial matrix. X-ray crystallography has recently provided us with the first atomic model of the bovine ADP/ATP carrier, which is a member of this family. The structure explains the typical three-fold sequence repeats and signature motif of mitochondrial carriers. However, the carrier was crystallised as a monomer in detergent, which is inconsistent with the consensus that mitochondrial carriers exist as homo-dimers. The projection structure of the yeast ADP/ATP carrier by electron crystallography shows that carriers could form homo-dimers in the membrane. PMID- 15111104 TI - L-type voltage-gated calcium channels: understanding function through structure. AB - L-type voltage-gated calcium channels (VGCCs) are multisubunit membrane proteins that regulate calcium influx into excitable cells. Within the last two years there have been four separate reports describing the structure of the skeletal muscle VGCC determined by electron microscopy and single particle analysis methods. There are some discrepancies between the structures, as well as reports for both monomeric and dimeric forms of the channel. This article considers each of the VGCC structures in terms of similarities and differences with an emphasis upon translation of data into a biological context. PMID- 15111105 TI - Structure of cation channels, revealed by single particle electron microscopy. AB - A large barrier in the way to obtaining high-resolution structures of eukaryotic ion channels remains the expression and purification of sufficient amounts of channel protein to carry out crystallization trials. In the absence of crystals, the main available source of structural information has been electron microscopy (EM), which is well suited to the visualization of isolated macromolecular complexes and their conformational changes. The recently published EM structures outline native conformations of eukaryotic cation channels that until now have eluded crystallization. According to these results, homo-tetrameric K(+) channels have a distinct two-layer architecture with connectors conjoining the two layers, while the pseudo-tetrameric Ca(2+) or Na(+) channels are more globular and have flexible surface loops, which makes the identification of subunits complicated. Subunits can be identified using atomic structure docking into the EM maps, labeling, or deletion studies. PMID- 15111106 TI - Critical assessment of a proposed model of Shaker. AB - Detailed three-dimensional structures at atomic resolution are essential to understand how voltage-activated K(+) channels function. The X-ray crystallographic structure of the KvAP channel has offered the first view at atomic resolution of the molecular architecture of a voltage-activated K(+) channel. In the crystal, the voltage sensors are bound by monoclonal Fab fragments, which apparently induce a non-native conformation of the tetrameric channel. Thus, despite this significant advance our knowledge of the native conformation of a Kv channel in a membrane remains incomplete. Numerous results from different experimental approaches provide very specific constraints on the structure of K(+) channels in functional conformations. These results can be used to go further in trying to picture the native conformation of voltage-gated K(+) channels. However, the direct translation of all the available information into three-dimensional models is not straightforward and many questions about the structure of voltage-activated K(+) channels are still unanswered. Our aim in this review is to summarize the most important pieces of information currently available and to provide a critical assessment of the model of Shaker recently proposed by Laine et al. PMID- 15111107 TI - ABC transporter architecture and mechanism: implications from the crystal structures of BtuCD and BtuF. AB - ABC transporters are ubiquitous membrane proteins that facilitate unidirectional substrate translocation across the lipid bilayer. Over the past five years, new crystal structures have advanced our understanding of how ABC transporters couple adenosine triphosphate (ATP) hydrolysis to substrate transport. In the following, we will briefly review the results of these structural investigations and outline their mechanistic implications. PMID- 15111108 TI - Heavier-than-air flying machines are impossible. AB - Many G protein-coupled receptor (GPCR) models have been built over the years. The release of the structure of bovine rhodopsin in August 2000 enabled us to analyze models built before that period to learn more about the models we build today. We conclude that the GPCR modelling field is riddled with 'common knowledge' similar to Lord Kelvin's remark in 1895 that "heavier-than-air flying machines are impossible", and we summarize what we think are the (im)possibilities of modelling GPCRs using the coordinates of bovine rhodopsin as a template. Associated WWW pages: www.gpcr.org/articles/2003_mod PMID- 15111109 TI - Evolution of photosystem I - from symmetry through pseudo-symmetry to asymmetry. AB - The evolution of photosystem (PS) I was probably initiated by the formation of a homodimeric reaction center similar to the one currently present in green bacteria. Gene duplication has generated a heterodimeric reaction center that subsequently evolved to the PSI present in cyanobacteria, algae and plant chloroplasts. During the evolution of PSI several attempts to maximize the efficiency of light harvesting took place in the various organisms. In the Chlorobiaceae, chlorosomes and FMO were added to the homodimeric reaction center. In cyanobacteria phycobilisomes and CP43' evolved to cope with the light limitations and stress conditions. The plant PSI utilizes a modular arrangement of membrane light-harvesting proteins (LHCI). We obtained structural information from the two ends of the evolutionary spectrum. Novel features in the structure of Chlorobium tepidum FMO are reported in this communication. Our structure of plant PSI reveals that the addition of subunit G provided the template for LHCI binding, and the addition of subunit H prevented the possibility of trimer formation and provided a binding site for LHCII and the onset of energy spillover from PSII to PSI. PMID- 15111111 TI - Automated large-scale purification of a G protein-coupled receptor for neurotensin. AB - Structure determination of integral membrane proteins requires milligram amounts of purified, functional protein on a regular basis. Here, we describe a protocol for the purification of a G protein-coupled neurotensin receptor fusion protein at the 3-mg or 10-mg level using immobilized metal affinity chromatography and a neurotensin column in a fully automated mode. Fermentation at a 200-l scale of Escherichia coli expressing functional receptors provides the material needed to feed into the purification routine. Constructs with tobacco etch virus protease recognition sites at either end of the receptor allow the isolation of neurotensin receptor devoid of its fusion partners. The presented expression and purification procedures are simple and robust, and provide the basis for crystallization experiments of receptors on a routine basis. PMID- 15111110 TI - The G protein-coupled receptor rhodopsin in the native membrane. AB - The higher-order structure of G protein-coupled receptors (GPCRs) in membranes may involve dimerization and formation of even larger oligomeric complexes. Here, we have investigated the organization of the prototypical GPCR rhodopsin in its native membrane by electron and atomic force microscopy (AFM). Disc membranes from mice were isolated and observed by AFM at room temperature. In all experimental conditions, rhodopsin forms structural dimers organized in paracrystalline arrays. A semi-empirical molecular model for the rhodopsin paracrystal is presented validating our previously reported results. Finally, we compare our model with other currently available models describing the supramolecular structure of GPCRs in the membrane. PMID- 15111112 TI - The plug domain of a neisserial TonB-dependent transporter retains structural integrity in the absence of its transmembrane beta-barrel. AB - Transferrin binding protein A (TbpA) is a TonB-dependent outer membrane protein expressed by pathogenic bacteria for iron acquisition from human transferrin. The N-terminal 160 residues (plug domain) of TbpA were overexpressed in both the periplasm and cytoplasm of Escherichia coli. We found this domain to be soluble and monodisperse in solution, exhibiting secondary structure elements found in plug domains of structurally characterized TonB-dependent transporters. Although the TbpA plug domain is apparently correctly folded, we were not able to observe an interaction with human transferrin by isothermal titration calorimetry or nitrocellulose binding assays. These experiments suggest that the plug domain may fold independently of the beta-barrel, but extracellular loops of the beta-barrel are required for ligand binding. PMID- 15111113 TI - Crystal structures of bR(D85S) favor a model of bacteriorhodopsin as a hydroxyl ion pump. AB - Structural features on the extracellular side of the D85S mutant of bacteriorhodopsin (bR) suggest that wild-type bR could be a hydroxyl-ion pump. A position between the protonated Schiff base and residue 85 serves as an anion binding site in the mutant protein, and hydroxyl ions should have access to this site during the O-intermediate of the wild-type bR photocycle. The guanidinium group of R82 is proposed (1) to serve as a shuttle that eliminates the Born energy penalty for entry of an anion into this binding pocket, and conversely, (2) to block the exit of a proton or a related proton carrier. PMID- 15111114 TI - The arrestin-bound conformation and dynamics of the phosphorylated carboxy terminal region of rhodopsin. AB - Visual arrestin binds to the phosphorylated carboxy-terminal region of rhodopsin to block interactions with transducin and terminate signaling in the rod photoreceptor cells. A synthetic seven-phospho-peptide from the C-terminal region of rhodopsin, Rh(330-348), has been shown to bind arrestin and mimic inhibition of signal transduction. In this study, we examine conformational changes in this synthetic peptide upon binding to arrestin by high-resolution proton nuclear magnetic resonance (NMR). We show that the peptide is completely disordered in solution, but becomes structured upon binding to arrestin. A control, unphosphorylated peptide that fails to bind to arrestin remains highly disordered. Specific NMR distance constraints are used to model the arrestin bound conformation. The models suggest that the phosphorylated carboxy-terminal region of rhodopsin, Rh(330-348), undergoes significant conformational changes and becomes structured upon binding to arrestin. PMID- 15111115 TI - Hemifluorinated surfactants: a non-dissociating environment for handling membrane proteins in aqueous solutions? AB - The instability of membrane proteins in detergent solution can generally be traced to the dissociating character of detergents and often correlates with delipidation. We examine here the possibility of substituting detergents, after membrane proteins have been solubilized, with non-detergent surfactants whose hydrophobic moiety contains a perfluorinated region that makes it lipophobic. In order to improve its affinity for the protein surface, the fluorinated chain is terminated by an ethyl group. Test proteins included bacteriorhodopsin, the cytochrome b(6)f complex, and the transmembrane region of the bacterial outer membrane protein OmpA. All three proteins were purified using classical detergents and transferred into solutions of C(2)H(5)C(6)F(12)C(2)H(4)-S-poly Tris-(hydroxymethyl)aminomethane (HF-TAC). Transfer to HF-TAC maintained the native state of the proteins and prevented their precipitation. Provided the concentration of HF-TAC was high enough, HF-TAC/membrane protein complexes ran as single bands upon centrifugation in sucrose gradients. Bacteriorhodopsin and the cytochrome b(6)f complex, both of which are detergent-sensitive, exhibited increased biochemical stability upon extended storage in the presence of a high concentration of HF-TAC as compared to detergent micelles. The stabilization of cytochrome b(6)f is at least partly due to a better retention of protein-bound lipids. PMID- 15111116 TI - Towards structure determination of neurotoxin II bound to nicotinic acetylcholine receptor: a solid-state NMR approach. AB - Solid-state magic-angle spinning nuclear magnetic resonance (NMR) has sufficient resolving power for full assignment of resonances and structure determination of immobilised biological samples as was recently shown for a small microcrystalline protein. In this work, we show that highly resolved spectra may be obtained from a system composed of a receptor-toxin complex. The NMR sample used for our studies consists of a membrane preparation of the nicotinic acetylcholine receptor from the electric organ of Torpedo californica which was incubated with uniformly 13C-,15N-labelled neurotoxin II. Despite the large size of the ligand receptor complex ( > 290 kDa) and the high lipid content of the sample, we were able to detect and identify residues from the ligand. The comparison with solution NMR data of the free toxin indicates that its overall structure is very similar when bound to the receptor, but significant changes were observed for one isoleucine. PMID- 15111117 TI - Computer simulation of the KvAP voltage-gated potassium channel: steered molecular dynamics of the voltage sensor. AB - The recent crystal structures of the voltage-gated potassium channel KvAP and its isolated voltage-sensing 'paddle' (composed of segments S1-S4) challenge existing models of voltage gating and raise a number of questions about the structure of the physiologically relevant state. We investigate a possible gating mechanism based on the crystal structures in a 10 ns steered molecular dynamics simulation of KvAP in a membrane-mimetic octane layer. The structure of the full KvAP protein has been modified by restraining the S2-S4 domain to the conformation of the isolated high-resolution paddle structure. After an initial relaxation, the paddle tips are pulled through the membrane from the intracellular to the extracellular side, corresponding to a putative change from closed to open. We describe the effect of this large-scale motion on the central pore domain, which remains largely unchanged, on the protein hydrogen-bonding network and on solvent. We analyze the motion of the S3b-S4 portion of the protein and propose a possible coupling mechanism between the paddle motion and the opening of the channel. Interactions between the arginine residues in S4, solvent and chloride ions are likely to play a role in the gating charge. PMID- 15111118 TI - Crystallographic analysis of AcrB. AB - A His-tagged derivative of the multidrug efflux pump AcrB could be crystallized in three different space groups (R3, R32 and P321). Experimental MAD-phasing maps from R32 AcrB(His) crystals were obtained to a resolution of 3.5 A. Datasets of native and substrate soaked AcrB(His) crystals were collected at the Swiss Light Source X06SA beamline up to a resolution of 2.7 A and refinement of these data provided good quality electron density maps, which allowed us to complement the published AcrB structure (PDB code 1iwg). Introduction of amino acids 860-865 and 868 lacking in the 1iwg structure and deletion of a highly disordered region (amino acids 669-678) improved R(free) and average B factors in the 2.7 A model. We could not identify significant densities indicating specific antibiotic binding sites in the AcrB R32 space group datasets under the soaking conditions tested. PMID- 15111119 TI - An antibody library for stabilizing and crystallizing membrane proteins - selecting binders to the citrate carrier CitS. AB - Co-crystallization of membrane proteins with antibody fragments may emerge as a general tool to facilitate crystal growth and improve crystal quality. The bound antibody fragment enlarges the hydrophilic part of the mostly hydrophobic membrane protein, thereby increasing the interaction area for possible protein protein contacts in the crystal. Additionally, it may restrain flexible parts or lock the membrane protein in a defined conformational state. For successful co crystallization trials, the antibody fragments must be stable in detergents during the extended period of crystal growth and must be easily produced in amounts necessary for crystallography. Therefore, we constructed a library of antibody Fab fragments from a framework subset of the HuCAL GOLD library (Morphosys, Munich, Germany). By combining the most stable and well expressed frameworks, V(H)3 and V(kappa)3, with the further stabilizing constant domains, a Fab library with the desired properties was obtained in a standard phage display format. As a proof of principle, we selected binders with phage display against the detergent-solubilized citrate transporter CitS of Klebsiella pneumoniae. We describe efficient methods for the immobilization of the membrane protein during selection, for ELISA screening, and for BIAcore evaluation. We demonstrate that the selected Fab fragments form stable complexes with native CitS and recognize conformational epitopes with affinities in the low nanomolar range. PMID- 15111120 TI - Cytochrome b5 oxidoreductase: expression and characterization of the original familial ideopathic methemoglobinemia mutations E255- and G291D. AB - NADH:cytochrome b5 oxidoreductase catalyzes the transfer of reducing equivalents from the physiological electron donor, NADH, to two molecules of cytochrome b5. Utilizing a heterologous expression system for the soluble, catalytic domain of the rat microsomal enzyme, we have produced two mutants, corresponding to E255- and G291D. These mutants correspond to the two specific mutations that were identified over a half century later following diagnosis of the original cases of type I recessive congenital methemoglobinemia (RCM). We have purified both the E255- and G291D variants to homogeneity to determine the molecular basis for type I RCM in these individuals. Both the E255- and G291D variants retained a full complement of FAD and exhibited absorption and CD spectroscopic properties comparable to those of the wild-type protein. Oxidation-reduction potentiometric titrations yielded standard midpoint potentials (E0') for the FAD/FADH2 couple of -271 and -273 mV for the E255- and G291D variants, respectively, which were comparable to the value of -268 mV obtained for the wild-type protein and confirmed that the redox potential of the flavin was unaffected by either mutation. Thermal and proteolytic stability studies revealed that while the G291D variant exhibited stability comparable to that of wild-type, the E255- variant was markedly less stable, indicative of an altered conformation. Initial-rate kinetic studies revealed that both mutants had decreased catalytic activity (kcat), with the E255- and G291D variants retaining approximately 38 and 58% of wild-type activity, respectively. However, the affinity for NADH (KmNADH) was decreased approximately 100-fold for E255- compared to only approximately 1.3 fold for G291D, results supported by the spectroscopic binding constant (Ks) obtained for G291D. These results indicate that the properties of both the E255- and G291D cytochrome b5 oxidoreductase mutants are similar to those of other variants that have been identified as resulting in the type I form of RCM. PMID- 15111121 TI - Rat cytochrome P450C24 (CYP24A1) and the role of F249 in substrate binding and catalytic activity. AB - A high level of functional recombinant rat cytochrome P450C24 enzyme (CYP24A1) was obtained (40-50mg/L) using an Escherichia coli expression system. Purified enzyme was stable with retention of spectral and catalytic activity. The rate of 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] side-chain oxidation and cleavage to the end-product calcitroic acid was directly related to the rate of electron transfer from the ferredoxin redox partner. It was determined from substrate induced spectral shifts that the 1 alpha- and 25-hydroxyl groups on vitamin D(3) metabolites and analogs were the major determinants for high-affinity binding to CYP24A1. Lowest K(d) values were obtained for 1 alpha-vitamin D(3) (0.06 microM) and 1,25-dihydroxyvitamin D(3) (0.05 microM) whereas unmodified parental vitamin D(3) and the non-secosteroid 25-hydroxycholesterol had lower affinities with K(d) values of 1.3 and 1.9 microM, respectively. The lowest binding affinity for natural vitamin D metabolites was observed for 24,25-dihydroxyvitamin D(3) [24,25(OH)(2)D(3)] (0.43 microM). Kinetic analyses of the two natural substrates 25-hydroxyvitamin D(3) [25(OH)D(3)] and 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] revealed similar K(m) values (0.35 and 0.38 microM, respectively), however, the turnover number was higher for 25(OH)D(3) compared to 1,25(OH)(2)D(3) (4.2 and 1 min(-1), respectively). Mutagenesis of F249 within the F-helix of CYP24A1 altered substrate binding and metabolism. Most notable, the hydrophobic to polar mutant F249T had a strong impact on lowering substrate binding affinity and catalysis of the final C(23) oxidation sequence from 24,25,26,27-tetranor-1,23-dihydroxyvitamin D(3) to calcitroic acid. Two other hydrophobic 249 mutants (F249A and F249Y) also lowered substrate binding and expressed metabolic abnormalities that included the C(23)-oxidation defect observed with mutant F249T plus a similar defect involving an earlier pathway action for the C(24) oxidation of 1,24,25-trihydroxyvitamin D(3). Therefore, Phe 249 within the F-helix was demonstrated to have an important role in properly binding and aligning substrate in the CYP24A1 active site for C(23) and C(24) oxidation reactions. PMID- 15111122 TI - Human dendritic cell lysosome-associated membrane protein expressed in lung type II pneumocytes. AB - Human dendritic cell LAMP (hDC-LAMP) is a unique member of the lysosome associated membrane protein (LAMP) family with a tissue distribution initially described as restricted to major histocompatibility class II (MHC II) compartments of activated DC before the translocation of MHC II to the cell surface [Immunity 9 (1998) 325]. In this report, we show that hDC-LAMP is also expressed by lung type II pneumocytes, another cell type with constitutive expression of MHC II. A recombinant hDC-LAMP protein and a monospecific anti-hDC LAMP polyclonal antibody were prepared. The antibody reacted specifically with hDC-LAMP sequences of hDC-LAMP protein expressed in transfected cells and with a 54 kDa protein of normal human lung tissue with properties corresponding to those of transgene expressed hDC-LAMP. Immunohistochemical analysis of hDC-LAMP in human lung showed its presence in alveolar type II epithelial cells (type II pneumocytes) as well as in cells in the interfollicular area of bronchus associated lymph nodes, where interdigitating DCs are concentrated, and with lesser staining of alveolar macrophages. The native protein contained approximately 16% carbohydrates, most of which are sialyl N-linked oligosaccharides, with an acidic isoelectric point (pI 4.8). The restricted localization of this protein to lung type II pneumocytes and DCs is in contrast to hLAMP-1, which was present in many cell types of the lung and lymph node. Type II pneumocytes are known to express MHC II and the abundant expression of hDC LAMP in these cells as well as in DCs suggests its possible relationship to specific MHC II related function(s) of DC and type II pneumocytes. PMID- 15111123 TI - Purification procedures determine the proteasome activation properties of REG gamma (PA28 gamma). AB - The proteasome activation properties of recombinant REG gamma molecules depend on purification procedures. Prior to ammonium sulfate precipitation recombinant REG gamma activates the trypsin-like catalytic subunit of the proteasome; afterwards it activates all three catalytic subunits. The expanded activation specificity is accompanied by reduced stability of the REG gamma heptamer providing support for the idea that a "tight" REG gamma heptamer suppresses the proteasome's chymotrypsin-like and postglutamyl-preferring active sites. In an attempt to determine whether REG gamma synthesized in mammalian cells also exhibits restricted activation properties, extracts were prepared from several mammalian organs and cell lines. Surprisingly, endogenous REG gamma was found to be largely monomeric. In an alternate approach, COS7 cells were cotransfected with plasmids expressing FLAG-REG gamma and REG gamma. The expressed FLAG-REG gamma molecules were shown to form oligomers with untagged REG gamma subunits, and the mixed oligomers preferentially activated the proteasome's trypsin-like subunit. Thus, REG gamma molecules synthesized in mammalian cells also exhibit restricted activation properties. PMID- 15111124 TI - The interpretation of multiple-step transient-state kinetic isotope effects. AB - In contrast to steady-state kinetic isotope effects (KIEs), transient-state tKIEs are both time and signal dependent and therefore require a very different form of theory for their interpretation. We have previously provided such a theory for the case of single-step isotopic substitutions. No such properly derived theory applicable to the analysis of multiple-step isotopic substitutions required by transient-state solvent isotope effect studies has been available up to this time. Here, we set forth a more general form of that theory which is applicable to multiple-step substituted cases. We prove three theorems: 1. the observed transient-state KIE for any given reactive component in the reaction sequence evaluated at zero time (tKIE(0)) is in fact the arithmetic product of the intrinsic KIEs of all the steps that precede the formation of that component. 2. The observed tKIE(0) is completely independent of the intrinsic KIEs of any reverse step in the reaction. 3. The intrinsic KIE of any step may be obtained by dividing the value of the tKIE(0) for that step by the value of the tKIE(0) of the immediately preceding step in the reaction sequence. PMID- 15111125 TI - Secondary structure, phospholipid membrane interactions, and fusion activity of two glutamate-rich analogs of influenza hemagglutinin fusion peptide. AB - Two synthetic mutants of influenza HA2 fusion peptide (residues 1-25), containing Glu on the polar (residues 4,8-E5(4,8)) or the hydrophobic (residues 3,7-E5(3,7)) face of the amphipathic helix, were synthesized and labeled with NBD at the N terminus. Introduction of Glu residues into the fusion peptide leads to increased sensitivity of various biochemical properties to pH compared to the wild type. The E5 peptides showed a decrease of alpha-helix content and increase of beta sheet structure. Lipid binding was diminished, but not abolished even at high pH. The E5 analogs penetrate the lipid bilayer less deeply than the wild type, especially at high pH. The N-terminal half of the peptide showed significant variation of the depth of the penetration into the lipid bilayer. Both E5 peptides were fusion active. The properties of E5(3,7) were more affected by the Glu substitution and showed greater variation with pH than E5(4,8). PMID- 15111126 TI - Flavoenzyme-catalyzed redox cycling of hydroxylamino- and amino metabolites of 2,4,6-trinitrotoluene: implications for their cytotoxicity. AB - The toxicity of 2,4,6-trinitrotoluene (TNT), a widespread environmental contaminant, is exerted through its enzymatic redox cycling and/or covalent binding of its reduction products to proteins and DNA. In this study, we examined the possibility of another cytotoxicity mechanism of the amino- and hydroxylamino metabolites of TNT, their flavoenzyme-catalyzed redox cycling. The above compounds acted as redox-cycling substrates for single-electron transferring NADPH:cytochrome P-450 reductase (P-450R) and ferredoxin:NADP(+) reductase (FNR), as well as substrates for the two-electron transferring flavoenzymes rat liver NAD(P)H:quinone oxidoreductase (NQO1) and Enterobacter cloacae NAD(P)H:nitroreductase (NR). Their reactivity in P-450R-, FNR-, and NR-catalyzed reactions increased with an increase in their single-electron reduction potential (E(1)(7)) or the decrease in the enthalpy of free radical formation. The cytotoxicity of the amino- and hydroxylamino metabolites of TNT towards bovine leukemia virus-transformed lamb kidney fibroblasts (line FLK) was partly prevented by the antioxidant N,N'-diphenyl-p-phenylene diamine and desferrioxamine, and potentiated by 1,3-bis-(2-chloroethyl)-1-nitrosourea, thus pointing to the involvement of oxidative stress. In general, their cytotoxicity increased with an increase in their electron accepting properties, or their reactivity towards the single-electron transferring FNR and P-450R. Thus, our data imply that the flavoenzyme-catalyzed redox cycling of amino and hydroxylamino metabolites of TNT may be an important factor in their cytotoxicity. PMID- 15111127 TI - Permeability characteristics and membrane affinity of flavonoids and alkyl gallates in Caco-2 cells and in phospholipid vesicles. AB - Biomembrane interactions of flavonoids and alkyl gallates were investigated using transport studies on Caco-2 cells and membrane affinity experiments in phospholipid vesicles. Flavone was rapidly absorbed across the cell monolayer (P(app),380 x 10(-6) cm/s), whereas efficient uptake but no apical to basolateral transport was observed with the flavonoids with higher degree of hydroxylation (e.g., quercetin and luteolin). The transport of alkyl gallates was governed by the length of the alkyl chain, i.e., methyl and propyl gallate were absorbed while octyl gallate showed cellular uptake but no transport. Flavonoids with several hydroxyl groups exhibited highest affinity for vesicle membranes, partition coefficients being 7.1 and 7.5 microM for luteolin and quercetin, respectively. In conclusion, the degree of hydroxylation, molecular configuration, and length of the side chain of flavonoids and alkyl gallates seem to have a highly important impact on their membrane affinity as well as on their permeability characteristics in Caco-2 cells. PMID- 15111128 TI - Chemical modification of muscle protein in diabetes. AB - Levels of glycation (fructose-lysine, FL) and advanced glycoxidation and lipoxidation end-products (AGE/ALEs) were measured in total skeletal (gastrocnemius) muscle and myofibril protein and compared to levels of the same compounds in insoluble skin collagen of control and diabetic rats. Levels of FL in total muscle and myofibril protein were 3-5% the level of FL in skin collagen. The AGE/ALEs, N(epsilon)-(carboxymethyl)lysine (CML) and N(epsilon) (carboxyethyl)lysine, were also significantly lower in total muscle and myofibril protein, approximately 25% of levels in skin collagen. The newly described sulfhydryl AGE/ALE, S-(carboxymethyl)cysteine (CMC), was also measured in muscle; levels of CMC were comparable to those of CML and increased similarly in response to diabetes. Although FL and AGE/ALEs increased in muscle protein in diabetes, the relative increase was less than that seen in skin collagen. These data indicate that muscle protein is partially protected against the increase in both glycation and AGE/ALE formation in diabetes. PMID- 15111129 TI - Modulation of gating currents of the Ca(v)3.1 calcium channel by alpha 2 delta 2 and gamma 5 subunits. AB - Modulatory effects of auxiliary alpha(2)delta(2) and gamma(5) subunits on intramembrane charge movement originating from the expressed Ca(v)3.1 calcium channel were investigated. Inward current was blocked by 1mM La(3+). Voltage dependences of Q(on) and Q(off), kinetics of ON- and OFF-charge movement, and I(max)/Q(max) ratio were measured in the absence and the presence of an auxiliary subunit. The alpha(2)delta(2) subunit accelerated significantly both ON- and OFF charge movement. I(max)/Q(max) ratio and Q(on)-V, Q(off)-V relations were not affected. Coexpression of the alpha(2)delta(2) subunit may accelerate channel transitions between individual closed states, but not the transition from the last closed channel state into an open state. Coexpression of the gamma(5) subunit accelerated the decay of the ON-charge transient and enhanced I(max)/Q(max) ratio. These effects suggest improvement of the coupling between the charge movement and the channel opening due to facilitation of transitions between individual closed states and the transition between the last closed state and an open state. PMID- 15111130 TI - Protein kinase B inhibits apoptosis induced by actinomycin D in ECV304 cells through phosphorylation of caspase 8. AB - Actinomycin D (act-D) anchors itself into DNA-base pairs by intercalation and thereby inhibits mRNA synthesis. It has been well established that act-D elicits apoptosis in various cell types involving endothelial cells. However, the regulatory mechanisms of actinomycin D-induced apoptotic cell death still remain unclear. Here, we investigated apoptotic cell death and its underlying regulatory mechanisms elicited by actinomycin D in ECV304. Act-D induced typical apoptotic features including chromatin condensation and translocation of phosphatidylserine. Since the phosphoinositide 3-OH kinase (PI3K)/protein kinase B (PKB) signaling pathway has been shown to prevent apoptosis in various cell types, it was of interest to determine if this pathway could protect against apoptosis induced by act-D. Inhibition of PI3K/PKB significantly increased act-D induced apoptosis. Moreover, act-D-induced cell death was physiologically linked to PKB-mediated cell survival through caspase-8. These results suggest that cross talk between the PKB and caspase-8 pathways may regulate the balance between cell survival and cell death in ECV304. PMID- 15111131 TI - 5'-Heterogeneity of mouse Dda3 transcripts is attributed to differential initiation of transcription and alternative splicing. AB - We have previously shown that mouse Dda3 gene is a p53 and p73 transcriptional target whose expression suppresses tumor cell growth. Here, we report the identification of multiple variants of Dda3 transcripts with diverse 5' sequences through 5'] rapid amplification of cDNA ends (5'-RACE) and RT-PCR. Analysis by primer extension and RNase protection revealed that the 5'-heterogeneity was generated by transcription initiation at multiple sites in exon 1 and intron 1 and by alternative splicing. These transcripts, both coding and non-coding, exhibited distinct expression patterns in various adult tissues and were developmentally regulated. Furthermore, they were induced in a p53-dependent manner by various stress signals. These data demonstrated that differential initiation of transcription and alternative splicing both participate in the regulation of Dda3 gene expression. PMID- 15111132 TI - Genome analyses of Streptomyces peucetius ATCC 27952 for the identification and comparison of cytochrome P450 complement with other Streptomyces. AB - We have determined the genome sequence of 8.7 Mb chromosome of Streptomyces peucetius ATCC 27952, which produces clinically important anthracycline chemotherapeutic agents of the polyketide class of antibiotics, daunorubicin and doxorubicin. The cytochrome P450 (CYP) superfamily is represented by 19 sequences in the S. peucetius. Among those, 15 code for functional genes, whereas the remaining four are pseudo genes. CYPs from S. peucetius are phylogenetically close to those of Streptomyces amermitilis. Four CYPs are associated with modular PKS of avermectin and two with doxorubicin biosynthetic gene cluster. CYP252A1 is the new family found in S. peucetius, which shares 38% identity to CYP51 from Streptomyces coelicolor A3 (2). Nine CYPs from S. peucetius are found in the cluster containing various regulatory genes including rar operon, conserved in S. coelicolor A3 (2) and Streptomyces griseus. Although two ferredoxins and four ferredoxin reductases have been identified so far, only one ferredoxin reductase was found in the cluster of CYP147F1 in S. peucetius. To date, 174 CYPs have been described from 45 Streptomyces species in all searchable databases. However, only 18 CYPs are clustered with ferredoxin. The comparative study of cytochrome P450s, ferredoxins, and ferredoxin reductases should be useful for the future development and manipulation of antibiotic biosynthetic pathways. PMID- 15111133 TI - Glucose starvation results in UDP-glucose deficiency and inactivation of glycogen synthase. AB - The effects of glucose starvation on glycogen synthase (GS) activity and protein expression were investigated. Fibroblasts were cultured in medium supplemented with either glucose or pyruvate. Pyruvate-cultured cells exhibited UDP-glucose contents that amounted to approximately 10% of those in cells cultured with glucose. GS activity, protein and mRNA amounts in pyruvate-cultured cells were decreased to approximately 35, 60, and 60%, respectively, of values in glucose cultured cells. Incubation of extracts from glucose-cultured cells with radioactive UDP-glucose resulted in substantial binding of ligand to immunoprecipitated GS. However, binding in immunoprecipitates from pyruvate cultured cells was decreased to approximately 25% of values in glucose-cultured cells. These data indicate that glucose starvation and the subsequent depletion of UDP-glucose result in: (1) inactivation of GS, owing to a decrease in its ability to bind UDP-glucose, and (2) decreased amount of GS protein, owing to a decrease in the levels of GS mRNA. PMID- 15111134 TI - Predictors of health-related quality of life after coronary artery bypass surgery. AB - BACKGROUND: Little is known about the determinants of health-related quality of life after coronary artery bypass surgery. We determined the predictors of overall physical and mental health status 6 months after the operation. METHODS: We evaluated 1,973 patients enrolled in a multicenter Veterans Affairs prospective cohort study who completed preoperative and 6-month postoperative Short Form-36 (SF-36) health status surveys. Multiple linear regression was used to identify the significant independent predictors of 6-month physical and mental component summary scores from the SF-36. RESULTS: In multivariable analyses adjusting for baseline health status, significant predictors of postoperative physical health status were a history of neurologic disease, peripheral vascular disease, chronic obstructive pulmonary disease, hypertension, current smoking, forced expiratory volume, left ventricular ejection fraction, and serum creatinine. Significant predictors of postoperative mental health status were a history of psychiatric disease, chronic obstructive pulmonary disease, current smoking, age, and New York Heart Association functional class. CONCLUSIONS: These predictors of health-related quality of life after coronary artery bypass surgery may be useful for preoperative risk assessment and counseling of patients with regard to anticipated health status outcomes. Factors such as current smoking and psychiatric disease may be targets for interventions to improve health-related quality of life outcomes. PMID- 15111135 TI - Does preoperative atrial fibrillation reduce survival after coronary artery bypass grafting? AB - BACKGROUND: Preoperative atrial fibrillation has been identified as a risk factor for reduced long-term survival after coronary artery bypass grafting. This study sought to determine whether atrial fibrillation is merely a marker for high-risk patients or an independent risk factor for time-related mortality. METHODS: From 1972 to 2000, 46,984 patients underwent primary isolated coronary artery bypass grafting; 451 (0.96% prevalence) had electrocardiogram-documented preoperative atrial fibrillation (n = 411) or flutter (n = 40). Characteristics of patients with and without atrial fibrillation were contrasted by multivariable logistic regression to form a propensity score. With this, comparable groups with and without atrial fibrillation were formed by pairwise propensity-matching to assess survival. RESULTS: Patients with preoperative atrial fibrillation were older (67 +/- 9.0 versus 59 +/- 9.8 years, p < 0.0001), had more left ventricular dysfunction (66% versus 52%, p < 0.0001) and hypertension (73% versus 59%, p < 0.0001), but less severe angina (39% moderate or severe versus 49%, p < 0.0001). Many of these factors are themselves predictors of increased time-related mortality. In propensity-matched patients, survival at 30 days and at 5 and 10 years for patients with versus without atrial fibrillation was 97% versus 99%, 68% versus 85%, and 42% versus 66%, respectively, a survival difference at 10 years of 24%. Median survival in patients with atrial fibrillation was 8.7 years versus 14 years for those without it. CONCLUSIONS: Atrial fibrillation in patients undergoing coronary artery bypass grafting is a marker for high-risk patients; in addition, atrial fibrillation itself substantially reduces long-term survival. Thus, if patients in atrial fibrillation require surgical revascularization, it is appropriate to consider performing a concomitant surgical ablation procedure. PMID- 15111136 TI - Surgical treatment of right coronary arteries with anomalous origin and slit ostium. AB - BACKGROUND: Right coronary arteries arising in the left sinus or ectopically in the anterior coronary sinus with slit ostium course inside the aorta. They are subject to variable systolic compression and can cause myocardial ischemia with its sequelae or death. METHODS: From May 1991 to March 2003, we treated 16 patients with anomalous origin of the right coronary artery from the left sinus and 4 whose right coronary artery arose ectopically in the anterior sinus. All patients had a slit ostium and underwent transaortic unroofing of the trunk to modify the proximal portion of the anomalous artery. RESULTS: All patients survived operation, although 1 patient died of unrelated causes. Nineteen patients were followed for a period from 0.2 to 11.8 years (median age, 53 years). One experienced angina 1 year after surgery and underwent percutaneous transluminal coronary angioplasty of a left internal thoracic to left anterior descending coronary artery anastomosis. All patients are New York Heart Association class I, without angina; none has sustained a myocardial infarction or required reoperation. CONCLUSIONS: Right coronary arteries that arise in anomalous fashion with a slit ostium can cause myocardial ischemia or death. Transaortic modification of the anomalous trunk addresses the anatomic and pathophysiologic features of the malformation that cause myocardial ischemia. Excellent results can be achieved with this surgical approach. PMID- 15111137 TI - Rationale for off-pump coronary revascularization to small branches--angiographic study of 1,283 anastomoses in 408 patients. AB - BACKGROUND: Off-pump coronary artery bypass grafting (OPCAB) has gained wide acceptance in tandem with the development of the stabilizer and associated operative techniques. However bypass grafting to the small branches of a beating heart is technically demanding and remains controversial. In the present study we evaluated the graft patency and quality of anastomoses to small coronary arteries by early postoperative angiography. METHODS: Between March 2000 and December 2002 a total of 1,328 anastomosed sites to coronary branches were studied angiographically in 404 patients representing 88.6% of all cases who underwent OPCAB in this period. The coronary artery branches were categorized as large (>1.5 mm, group L: 1,028 anastomoses sites) or small (< 1.5 mm, group S: 300 sites) by intraoperative measurement. As in situ grafts the internal thoracic artery (ITA) and the gastroepiploic artery (GEA) were used at 504 and 28 distal anastomosis sites respectively. The radial artery (RA) was used as a composite graft for 739 distal anastomosis sites. Sequential bypass grafting was performed at 388 anastomosis sites in side-to-side fashion. Arterial grafts were used in 96.1% of total bypass grafting. RESULTS: The percentage of male gender was 78.3% in group S and 87.2% in group L (p = 0.025). The ITA was used in 43.7% of group L and 18.3% of group S (p < 0.0001). The RA was used in 49.4% of group L and 77% of group S (p < 0.0001). The overall patency and stenosis free rates (FitzGibbon Type A) were 97.2% and 96.2%. Graft patency and stenosis free rates in group S (96.7% and 93.3%) were as good as those in group L (97.5% and 97.1%). In group S, the patency and stenosis free rates of SV grafts were 71.4% and 57.1%. On the other hand, those of ITA grafts were 100% and 98.3% (p = 0.53 vs. saphenous vein graft [SVG]) and RA grafts were 95.8% and 92.1% (p = 0.61 vs. SVG) respectively. In group S, the graft patency and stenosis free rates of bypass to the obtuse marginal (OM) (93.7% and 87.5%) were slightly lower than those to other implantation sites left anterior descending (LAD: 100% and 97.3%; PL: 96.5% and 92.3%; DI: 98.0% and 96%; PDA: 97.0% and 97.0%; right coronary artery [RCA]: 100% and 100%) although there was no statistical significance. The graft patency and stenosis free rates were slightly better with side-to-side anastomosis than with end-to-side anastomosis (side-to-side: 98.1% and 95.8% vs. end-to-side 96.3% and 86.3%) in group S. CONCLUSIONS: OPCAB to small coronary artery branches with arterial grafts provided satisfactory graft patency and stenosis free rates. PMID- 15111138 TI - Exercise therapy after coronary artery bypass graft surgery: a randomized comparison of a high and low frequency exercise therapy program. AB - BACKGROUND: Postoperative exercise therapy aims at recovering, as soon as possible, independence in the basic physical activities; but the type, intensity, and therefore the costs of the programs, vary widely. The aim of this study was to compare the effectiveness of a low frequency (once daily, not in the weekend) program with a high frequency (twice daily, including the weekend) one and to assess whether the latter would yield sufficient benefit for the patient to justify higher costs in material and personnel (physiotherapists) after uncomplicated coronary artery bypass graft (CABG) surgery. METHODS: Two-hundred and forty-six patients were randomly allocated to either a low or high frequency exercise program. Endpoints were the functional level as measured by the achievement of five activity milestones, the patient's independence (functional independence measures [FIM]) as assessed by a structured interview, the amount of daily physical activity (activity monitor), and patient satisfaction (questionnaire). Except for patient satisfaction, all measurements were done in the first week after surgery. RESULTS: Patients with the high frequency exercise program achieved functional milestones faster than patients with the low frequency exercise program (p = 0.007). The frequency of the exercise program had no influence on functional independence as measured with the FIM or quantity of physical activity. The satisfaction degree was greater in the high frequency group (p = 0.032), although the low frequency group was not dissatisfied. CONCLUSIONS: A high frequency exercise program leads to earlier performance of functional milestones and yields more satisfaction after uncomplicated CABG surgery and this should lead to an earlier discharge. On the other hand, if the shortage of physiotherapists remains unchanged or even increases, the low frequency program also yields excellent functional results, albeit at the cost of a somewhat longer hospital stay: but it would allow a sensible redistribution of the physiotherapists activity towards complicated and, therefore, more demanding patients. PMID- 15111139 TI - First-year outcomes of beating heart coronary artery bypass grafting using proximal mechanical connectors. AB - BACKGROUND: To determine the extended results of mechanical connectors we compared the 1-year outcomes of patients having beating heart coronary artery bypass surgery with at least one sutured or mechanically connected proximal vein graft anastomosis. METHODS: From May 2001 to December 2001, 166 patients were identified as having undergone off-pump bypass grafting utilizing at least one St. Jude symmetry aortic connector (St Jude Medical Anastomotic Technology Group, St. Paul, MN). Follow-up for major adverse cardiac events (MACEs), which is defined as cardiac mortality, myocardial infarction, or revascularization of a previous target vessel, was obtained on 162 patients (97.6%). A control group of 159 patients was identified from a cohort of patients having beating heart surgery with one or more sutured proximal vein graft anastomosis in the preceding year. The MACE follow-ups were obtained in 136 patients (85.6%) by direct telephone contact. RESULTS: Patients with connectors showed an accelerated number of MACEs beginning approximately 180 days from the time of surgery and stabilizing at approximately 300 days. Logistic regression analysis identified the presence of diabetes as a significant preoperative risk factor predisposing patients to earlier onset of MACEs (p = 0.03) with an odds ratio of 2.9 (95% confidence interval, 1.1 to 7.6). Insulin dependent diabetics showed no differences between connector and control patients in the frequency or timing of MACEs. Connector patients using oral hypoglycemic agents demonstrated a significant deviation (p = 0.01) from a similar control population in the prevalence and timing of MACEs. CONCLUSIONS: Connector patients showed an increased incidence of early MACEs. These events were characterized by an increased requirement for early target vessel revascularization and were predominantly in noninsulin-dependent diabetics. PMID- 15111140 TI - Skeletonized radial artery graft with the St. Jude Medical Symmetry Bypass System (aortic connector system). AB - BACKGROUND: We report our initial experience with an automatic anastomotic device using skeletonized radial artery in patients requiring off-pump coronary artery bypass grafting (CABG). METHODS: St. Jude Medical, Inc, Symmetry Bypass System (aortic connector system [ACS]) (St Jude Medical, St. Paul, MN) was used in ten patients. Ten consecutive patients who underwent off-pump CABG and who received at least one radial artery graft proximal anastomosis using the ACS were evaluated. The radial artery (RA) was harvested in a skeletonized fashion and applied to the ACS in the same manner as applying saphenous vein graft. The creation of the anastomosis lasted no longer than a few seconds. RESULTS: Our attempt to use the ACS for proximal anastomosis of the RA was successful in all ten patients. Mean operating time was 3.2 +/- 0.6 minutes and an average of 3.0 +/- 0.9 bypass grafts (range, 2 to 5 grafts) were performed. There was no postoperative fatal complication. Postoperative angiographic control showed that all grafts were widely patent including grafts other than the RA. During the mean postoperative follow-up of 10.3 +/- 2.9 months, there was no cardiac-related event in any patient. CONCLUSIONS: The St. Jude Medical Symmetry aortic connector system allows the construction of uniform and widely patent anastomoses in RA graft and does not require aortic side biting. Skeletonization of the RA is a safe and effective technique for applying ACS in off-pump CABG using multiple arterial grafts. PMID- 15111142 TI - Safety and efficacy of perioperative cell salvage and autotransfusion after coronary artery bypass grafting: a randomized trial. AB - BACKGROUND: The aim of this study was to ascertain whether cell salvage and autotransfusion after first time elective coronary artery bypass grafting is associated with a significant reduction in the use of homologous blood, a clinically significant derangement of postoperative clotting profiles, or an increased risk of postoperative bleeding. METHODS: Patients were randomized to autotransfusion (n = 98) receiving autotransfused washed blood from intraoperative cell salvage and postoperative mediastinal fluid cell salvage after coronary artery bypass surgery or control (n = 102) receiving stored homologous blood only after coronary artery bypass surgery. RESULTS: There was no statistical difference between the groups in terms of demographics, comorbidity, risk stratification, or operative details. Mean volume of blood autotransfused was 367 +/- 113 mL. Patients in the autotransfusion group were significantly less likely to receive a homologous blood transfusion compared with controls (odds ratio 0.40, 95% confidence interval [CI] 0.22-0.71) and received significantly fewer units of blood per patient compared with controls (0.43 +/- 1.5 vs 0.90 +/- 2.0 U, p = 0.02). There was no difference between the groups in terms of postoperative blood loss, fluid requirements, blood product requirements, or in the incidence of adverse clinical events (p = NS chi(2)). Autotransfusion did not produce any significant derangement of thromboelastograph values or laboratory measures of clotting pathway function (prothrombin time, activated partial thromboplastin time, fibrinogen, and fibrinogen D-dimer levels) when compared with the effect of homologous blood transfusion (p = NS, repeated measures analysis of variance [MANOVA]). CONCLUSIONS: Autotransfusion is a safe and effective method of reducing the use of homologous bank blood after routine first time coronary artery bypass grafting. PMID- 15111143 TI - Off-pump coronary artery bypass operation does not increase procoagulant and fibrinolytic activity: preliminary results. AB - BACKGROUND: This study analyzes the effects on coagulation and fibrinolysis comparing off-pump coronary artery bypass (OPCAB) and on-pump CABG operations. METHODS: In a prospective, nonrandomized, comparative evaluation, patients scheduled for elective myocardial revascularization were studied. Due to possible confounding factors patients with postoperative retransfusion of mediastinal shed blood were excluded. Nine patients underwent OPCAB operation and 16 underwent on pump CABG. Activated clotting time (ACT) was adjusted to 250 seconds in OPCAB (81 +/- 18 [mean +/- SD] IU/kg heparin) and to more than 480 seconds in on-pump CABG (400 IU/kg heparin, additional 10,000 IU in pump prime). Perioperatively blood samples were collected and hematologic and hemostatic variables including fibrinopeptide A (FPA), fibrin monomer (FM), thrombin-antithrombin complex (TAT), and D-dimer were analyzed. RESULTS: Both groups showed comparable demographic variables. Number of grafts per patient was slightly higher in the on-pump group (3.6 +/- 0.6 versus 3.0 +/- 1.1, p = 0.23). The FPA levels did not differ significantly between the groups. The FM, TAT, and D-dimer values were significantly higher in on-pump CABG (p < 0.0001, p < 0.01, and p < 0.0001, respectively), reflecting increased coagulant and fibrinolytic activity. This was also the case when values were corrected for hemodilution. CONCLUSIONS: Despite lower systemic anticoagulation activation of coagulation and fibrinolysis is reduced in OPCAB compared with on-pump CABG. Reduced thrombin generation and reduced fibrinolytic activity in OPCAB indicates better preservation of hemostasis. We suggest the term "preserved hemostasis" instead of "hypercoagulant activity" with respect to OPCAB. PMID- 15111144 TI - Do off-pump techniques reduce the incidence of postoperative atrial fibrillation in elderly patients undergoing coronary artery bypass grafting? AB - BACKGROUND: Atrial fibrillation is the most common postoperative complication in patients undergoing coronary artery bypass grafting (CABG) with advancing age having been shown to have a significant association with its incidence. This study aims to assess whether off-pump coronary artery bypass (OPCAB) reduces the incidence of atrial fibrillation in elderly patients. METHODS: A meta-analysis of all observational studies reporting a comparison between the two techniques in elderly patients (> 70 years) between 1999-2003 was performed. The primary outcome of interest was the incidence of postoperative atrial fibrillation. The quality of each study was evaluated by examining three items: patient selection, matching of the off-pump and cardiopulmonary bypass patient groups, and assessment of outcome. Meta-regression analysis was undertaken to see the effects of study size and quality on the calculated odds ratio. RESULTS: Eight studies fulfilled our inclusion criteria, all of which were nonrandomized. In total the studies identified 3017 subjects, of which 764 had off-pump surgery (25%) and 2253 underwent cardiopulmonary bypass (75%). Meta-analysis showed that after off pump surgery there was a significantly lower incidence of postoperative atrial fibrillation in these patients (odds ratio 0.70, 95% confidence interval [CI] 0.56-0.89). Meta-regression analysis including study characteristics did not show any associations affecting the calculated odds ratio of atrial fibrillation. CONCLUSIONS: Our study demonstrates a reduced incidence of postoperative atrial fibrillation in an elderly population with off-pump as compared with cardiopulmonary bypass techniques. We appreciate, however, that our statistical analysis uses nonrandomized published data and that the results must be treated with caution. If this finding is confirmed by a large-scale randomized trial, it has significant implications on the operative strategy employed for this patient group. PMID- 15111145 TI - Diseased vein grafts express elevated inflammatory cytokine levels compared with atherosclerotic coronary arteries. AB - BACKGROUND: The pathologic modifications characterizing vein graft disease resemble those observed in native arteriosclerosis, but in accelerated form. Although both disorders are considered to be inflammatory diseases, it remains to be determined whether diseased vein grafts and atherosclerotic coronary arteries differentially express inflammatory mediators. Therefore, we examined whether differences in the expression of proinflammatory cytokines by these two distinct vascular pathologies favor the accelerated inflammation within diseased vein grafts. METHODS: The messengerRNA expression of various cytokines (interleukin-1 beta [IL-1 beta], IL-6, IL-8, tumor necrosis factor-alpha [TNF-alpha], interferon gamma [IFN-gamma]) was quantified using real-time reverse transcriptase polymerase chain reaction (RT-PCR) in tissue samples of native saphenous veins (NSV, n = 5), diseased coronary arteries (CAD, n = 25), and diseased vein grafts (VG, n = 13). RESULTS: Native saphenous veins did not contain any detectable transcripts except for IFN-gamma. As expected, CAD was characterized by the expression of IL-1 beta, IL-6, IL-8, IFN-gamma, and TNF-alpha mRNA. Interestingly VG also expressed these mediators, but at markedly higher levels. Quantification by RT-PCR revealed that, compared with specimens from the CAD group, VG specimens contained 5.8 +/- 1.2 times, 286 +/- 22 times, and 29 +/- 7.3 times as many transcripts for the cytokines IL-1 beta, IL-6 and TNF-alpha, respectively, as well as 25 +/- 8.3 times more transcripts for the chemokine IL-8. In contrast, the expression of IFN-gamma transcripts did not differ among the groups. CONCLUSIONS: The elevated expression of proinflammatory cytokine transcripts supports the hypothesis that diseased vein grafts, compared with atherosclerotic coronary arteries, are characterized by enhanced inflammatory activity that might accelerate atherosclerotic modifications. This may implicate new therapeutic strategies for the prevention of vein graft disease. PMID- 15111146 TI - Local application of rapamycin inhibits neointimal hyperplasia in experimental vein grafts. AB - BACKGROUND: Rapamycin is an immunosuppressive agent which also exhibits marked antiproliferative properties. Rapamycin coated stents have been demonstrated to suppress restenosis in experimental and clinical studies of percutaneous coronary catheter intervention. We investigated whether rapamycin can reduce neointima formation in a mouse model of vein graft disease. METHODS: C57BL6J mice underwent interposition of the inferior vena cava from isogenic donor mice into the common carotid artery using a previously described cuff technique. In the treatment group, 100 microg or 200 microg of rapamycin was applied locally in pluronic gel. The control group did not receive local treatment. Grafts were harvested at 1, 2, 4, and 6 weeks and underwent morphometric analysis as well as immunohistochemical analysis. RESULTS: In grafted veins without treatment (controls), median intimal thickness was 9.6 (6.4 to 29)microm, 11.9 (7.9 to 39.9)microm, 46.6 (12.4 to 57.7)microm, and 57.5 (32.5 to 71.1)microm after 1, 2, 4, and 6 weeks, respectively. Treatment with 100 microg or 200 microg rapamycin showed a dose dependent reduction of intimal thickness. In the 200 microg rapamycin treatment group the intimal thickness was 4.3 (3.4 to 5.6)microm, 3.8 (3.2 to 6.3)microm, 17.1 (4.8 to 63)microm, and 33.9 (11.3 to 80.3)microm after 1, 2, 4, and 6 weeks, respectively. This difference of intimal thickness of 200 microg treated animals compared with controls was statistically significant at 1 and 2 weeks. Immunohistochemically the reduction of intimal thickness was associated with a decreased amount of infiltration of CD-8 positive cells and a decreased amount of metallothionein positive cells in the rapamycin treated grafts. CONCLUSIONS: We conclude that perivascular application of rapamycin inhibits neointimal hyperplasia of vein grafts in a mouse model. These results suggest that rapamycin may have a therapeutic potential for the treatment of vein graft disease. PMID- 15111147 TI - Endoscopic localization and assessment of coronary arteries by 13 MHz epicardial ultrasound. AB - BACKGROUND: In totally endoscopic coronary artery bypass grafting the target coronary artery is difficult to locate and assess. We explored the capacity of a high-frequency epicardial ultrasound mini-transducer (Aloka, Tokyo, Japan) to endoscopically locate and assess the left anterior descending (LAD), third obtuse marginal (OM3), and right posterior descending (RDP) coronary arteries. METHODS: In eight pigs, the LAD, OM3, and RDP were endoscopically exposed. The mini transducer was manipulated by the "da Vinci" telemanipulation system (Intuitive Surgical, Inc, Mountain View, CA) over the unstabilized and stabilized epicardium to identify the target artery, obtain a scout scan, and both transverse and longitudinal images. RESULTS: In both unstabilized and stabilized conditions, the LAD and RDP were identified within a median of 29 seconds. In stabilized conditions, assessment was complete in 112 seconds (92 to 205) (median with range) for the LAD and 140 seconds (54 to 197) for the RDP. Stabilization of the OM3 was required for identification (16 [5 to 60]) and assessment (111 [82 to 225]). Overall identification was correct in 23 of 24 arteries. The OM branches and RDP became fully exposed endoscopically with stroke volume (SV) and mean arterial pressure (MAP) remaining at 67% +/- 11% (mean +/- standard error of the mean) and 70% +/- 5% of baseline values, respectively. Scanning itself did not augment the decrease in SV and MAP significantly. CONCLUSIONS: After proper endoscopic exposure and stabilization, robot-assisted epicardial ultrasound scanning enabled endoscopic identification and assessment of major coronary arteries within a median of 169 seconds per artery. Exposure, stabilization, and scanning were accompanied by an acceptable drop in stroke volume and mean arterial pressure. PMID- 15111148 TI - Effect of prior malignancy on survival after cardiac surgery. AB - BACKGROUND: The number of patients with a previously treated tumor, needing cardiac surgery is increasing. Whether this operation in these patients is justified is determined by the long-term outcome. METHODS: Of 8620 patients referred for cardiac surgery, 205 had a documented malignant tumor. The time interval between the occurrence of the tumor and the cardiac surgery was recorded. These patients were matched with 205 patients without a tumor according to age, gender, comorbidity and type of cardiac surgery. The patients were followed retrospectively. A chi(2) Kaplan Meier and Cox' regression analysis were performed. RESULTS: During follow-up, 95.8% of the patients were traced (2794 patient years). Univariate analysis showed that 5- and 10-year survival was better in patients without a malignant tumor in the history (0.91 +/- 0.02 versus 0.72 +/- 0.03 and 0.73 +/- 0.04 versus 0.40 +/- 0.05; p < 0.0001). For shorter time intervals, mortality for all causes and mortality due to the tumor increase significantly (p < 0.0001). Multivariate analysis identified 4 independent variables: a malignant tumor in the history (p < 0.001), chronic obstructive pulmonary disease (p = 0.003), age (p = 0.001), and impaired left ventricular function (p = 0.035) CONCLUSIONS: A malignant tumor in the history is the most prognostic factor after cardiac surgery, but the operation is still rewarding. Fatal progression of the tumor is seen if the time interval between the occurrence of the malignant tumor and cardiac surgery is short. Other unfavorable factors are decreased left ventricular function, chronic obstructive pulmonary disease and high age. PMID- 15111150 TI - Edge-to-edge (Alfieri) mitral repair: results in diverse clinical settings. AB - BACKGROUND: Complex mitral regurgitation (MR) jets can make repair challenging; edge-to-edge (Alfieri) repair augments the repertoire of repair techniques. Objectives of this study were to demonstrate causes of MR amenable to edge-to edge repair and to determine safety, obstructive potential, and durability of edge-to-edge repair. METHODS: From January 1997 to October 2001, 224 patients underwent Alfieri repair. Indications included ischemic cardiomyopathy (n = 143, 64%), myxomatous disease (n = 31, 14%), dilated cardiomyopathy (n = 27, 12%), and hypertrophic obstructive cardiomyopathy (n = 14, 6%). Concomitant ring annuloplasty was performed in 188 patients (84%). Two additional patients had takedown of an Alfieri repair in the operating room for obstruction. Preoperative MR was 4+ in 109 patients (50%) and 3+ in 65 (30%). Postoperative and follow-up mitral gradient and return of MR were assessed using 396 transthoracic echocardiograms and longitudinal analyses. RESULTS: Hospital mortality was 2% (5 of 224). Mitral valve mean gradient was low (3.7 mm Hg) and nonprogressive (p = 0.7), although peak gradient rose slightly, from mean 8.4 to 10.0 mm Hg (p = 0.01). During the first 3 postoperative months, absence of MR declined to 40%, and prevalence of 3+ MR increased to 14%, then rose slowly thereafter. Fourteen patients--12 within 2 years--underwent mitral valve reoperation, none for stenosis; 7 patients--6 within 2 years--underwent heart transplantation. CONCLUSIONS: Alfieri mitral repair can be used in a variety of settings with a low risk of creating mitral stenosis. However, in ischemic MR, steadily increasing prevalence of moderately severe and severe regurgitation after edge-to edge repair suggests other techniques are needed. PMID- 15111151 TI - Biological or mechanical prostheses in tricuspid position? A meta-analysis of intra-institutional results. AB - BACKGROUND: Tricuspid valve replacement (TVR) is an uncommon procedure. The use of biological vs mechanical prostheses in TVR has pros and cons. Therefore, we debate the choice between the different types of valves by means of a meta analysis of studies of the last decade. METHODS: The heading "tricuspid valve replacement and (bio* or mec*)" was used to retrieve studies from Medline, Current Contents, and Embase. Eight out of 11 studies met the preset strict criteria: intra-institutional comparison of results of biological or mechanical TVR. Survival of hospital-discharged patients was recalculated to reduce the effect of unbalanced perioperative risk factors on overall survival. Hazard ratio was obtained from actuarial survival graphics comparison and at-risk groups, according to the method described by Parmar. If missing, the number of patients at risk was approximated assuming constant and noninformative censoring. Hazard pooling was done according to study heterogeneity. Bioprostheses were assumed as the gold standard and mechanical prostheses assumed as the challenging device. Therefore, a hazard more than 1 pointed to a higher risk of mechanical prostheses. Our 1998 study was updated for this analysis. RESULTS: In this study, 1,160 prostheses and 6,046 follow-up years were analyzed. The pooled survival hazard ratio of mechanical prostheses versus bioprostheses was 1.07 (0.84 to 1.35, p = 0.60). The pooled freedom from reoperation hazard ratio was 1.24 (0.67 to 2.31, p = 0.67). Pooled survival differences were trivial, favoring mechanical prostheses at 1 (-0.04%) and 15 years (-1.1%) and favoring bioprostheses (+1.8%) at 10 years. CONCLUSIONS: There is not a gold standard in tricuspid prostheses replacement. Prosthetic choice is left to the surgeon's clinical judgment, taking into consideration each patient's characteristics and needs. PMID- 15111152 TI - High-intensity transient signals due to prosthetic valve obstruction: diagnostic and therapeutic implications. AB - BACKGROUND: High-intensity transient signals (HITS) can be detected by transcranial Doppler ultrasonography (TCD) in patients carrying a mechanical prosthetic valve. The HITS counts and a frequency analysis were evaluated in patients with prosthetic valve obstruction in the aortic position. METHODS: Simultaneous echocardiographic, cineradiographic, and TCD evaluations for a St. Jude Medical valve were performed in 108 patients. All patients were asymptomatic and had no significant stenosis of the carotid artery. The HITS were identified according to criteria established by consensus of the International Cerebral Hemodynamics Symposium. RESULTS: The HITS counts in 69 patients with normal prosthetic valve function were 2.2 +/- 4.4, and the counts in 39 patients with prosthetic valve obstruction (group D) were 8.3 +/- 10.8. This difference was significant (p = 0.0002). In 2 patients of group D who had a greater rate of less than 400 Hz HITS, which were produced by solid microemboli, thrombolysis resulted in a mitigation of restricted leaflet movement equal to or greater than 10(o). The total number of HITS decreased and the rate of less than 400 Hz HITS also markedly decreased after thrombolysis in these 2 patients. On the other hand, 4 patients who obtained no improvement of leaflet movement by thrombolysis had lower rates of less than 400 Hz HITS than did the 2 patients. CONCLUSIONS: These results suggest that measurement of HITS counts is useful for detection of prosthetic valve obstruction, and that a frequency analysis of HITS may be valuable to clarify the cause of the obstructed prosthetic valves. PMID- 15111153 TI - Acute type B aortic dissection in elderly patients: clinical features, outcomes, and simple risk stratification rule. AB - BACKGROUND: The clinical features and outcomes of elderly patients with acute type B aortic dissection (ABAD) are less well known. Accordingly, we sought to evaluate the clinical features and outcomes and derive a simple risk stratification rule for elderly with ABAD. METHODS: We categorized 383 patients with ABAD enrolled in the International Registry of Acute Aortic Dissection into two strata (aged less than 70 years and aged 70 years or more) and compared their clinical features and in-hospital outcomes. Further, we developed a clinical decision rule to risk-stratify elderly with ABAD. RESULTS: Forty-two percent (161 of 383) of patients with ABAD were aged 70 years or more. Hypertension, diabetes, history of prior aortic aneurysm, and arteriosclerosis were more common in the elderly patients, whereas Marfan syndrome and cocaine abuse were less common. The in-hospital complication of hypotension/shock was more common among elderly, and malperfusion of a visceral organ less frequent among elderly patients. In hospital mortality was higher in the elderly cohort compared with the younger patients (16% versus 10%, p = 0.07). A classification tree identified that elderly patients with hypotension/shock had the highest risk of death (56%). In absence of this, any branch vessel involvement was associated with the next highest mortality rate (28.6%) followed by presence of periaortic hematoma (10.5%). In contrast, elderly patients without any of these three risk factors had an extremely low mortality rate (1.3%). CONCLUSIONS: Our study highlights important differences between older and younger patients with ABAD in their clinical characteristics, management, and outcomes. We also propose a simple decision rule that allows stepwise risk-stratification in elderly patients with ABAD. PMID- 15111155 TI - Neurocognitive outcome after retrograde cerebral perfusion. AB - BACKGROUND: Neurocognitive outcome in patients undergoing thoracic aortic surgery using retrograde cerebral perfusion (RCP) remains uncertain. METHODS: Forty-two patients undergoing thoracic aortic surgery using RCP were enrolled in the study. The patients' neurocognitive state was assessed by the revised Wechsler Adult Intelligence Scale (WAIS-R) a few days before operation, at 2 to 3 weeks, and at 4 to 6 months after operation. RESULTS: There were no stroke, seizure, and hospital death. Significant performance deterioration was observed in digit span, arithmetic, and picture completion of the WAIS-R subtests. Bivariate comparison showed that older age (late vocabulary, late similarities, and late object assembly), longer RCP time (early picture arrangement, and early block design), later awake time (early and late picture arrangement, and early block design), longer respirator use (early and late digit span, late picture arrangement), longer ICU time (late picture completion, early and late picture arrangement, and early block design), and longer hospital stay (early picture arrangement) were significantly associated with the decline in neurocognitive performance. Stepwise logistic regression analysis disclosed that older age (late similarities and late object assembly), later awake time (late picture arrangement), and longer respirator use (early and late digit span, and late picture arrangement) were most predictive for the decline in neurocognitive performance. CONCLUSIONS: On average, digit span, arithmetic, and picture completion tests were most sensitive in detecting the decline in neurocognitive performance. The relationship between the duration of RCP and neurocognitive test results was not significant. PMID- 15111157 TI - Evaluation of native valve-sparing aortic root reconstruction with direct imaging -reimplantation or remodeling? AB - BACKGROUND: Aortic root reimplantation and remodeling have been used to preserve the native aortic valve. However, direct observation of valve motions with these techniques has not been performed. METHODS: Mongrel dogs were studied. The beating heart model was created using modified Tyrode's solution. Normal aortic valves and aortic valves preserved with the remodeling or reimplantation procedure were observed with an endoscope, and behavior was recorded on a high speed video (200 frames/s). The aortic valve orifice area was measured at 11 data points per beat. A predictable maximum valve orifice area was defined as an area encircled by the three commissures. A ratio of each aortic valve orifice area to the predictable maximum valve orifice area was calculated. The control group, the reimplantation group, and the remodeling group were compared. RESULTS: The preserved aortic valve with reimplantation showed bending and asymmetric motion. The ratio of aortic valve orifice area and predictable maximum valve orifice area in the reimplantation group was significantly smaller compared with the control and remodeling groups. CONCLUSIONS: The opening and closing behavior of the aortic valve preserved with the reimplantation procedure was impaired. It was speculated that the remodeling procedure may preserve more physiologic root function compared with the reimplantation procedure. PMID- 15111158 TI - Initial experience with miniature axial flow ventricular assist devices for postcardiotomy heart failure. AB - BACKGROUND: The recently introduced Impella Recover (Impella CardioSystems AG, Aachen, Germany) microaxial flow left and right ventricular assist devices (LVAD/RVAD) were evaluated as they provide circulatory support in the setting of postcardiotomy heart failure refractory to high-dose inotropic and intra-aortic balloon pump (IABP) support. METHODS: Between May 2002 and November 2002, the Recover LVAD was implanted in six patients (64 +/- 11 years) with acute left heart failure following coronary artery bypass procedures. Preoperative left ventricular (LV) ejection fraction was compromised (28% +/- 12%, 12% to 45%). Three patients presented with unstable circulation or cardiogenic shock following acute myocardial infarction, with a predicted mortality rate of 44% +/- 11% (EuroSCORE). Intraoperatively, severe heart failure was present with a more than 70% mortality rate predicted by the IABP score. The Recover RVAD and LVAD were combined to provide biventricular assist device (BVAD) support in one case of post-transplant graft failure. RESULTS: The Recover LVAD delivered blood flows of up to 5 L/min. A moderate degree of hemolysis and a reduction in platelet count were noted. Four patients were weaned from the LVAD after 169 +/- 34 hours, two of whom remain long-term survivors. Full recovery of graft function allowed weaning of the patient from BVAD support after six days. CONCLUSIONS: The initial experience with the Impella Recover VADs proved the new systems to be advantageous regarding the ease of implantation and device removal, low anticoagulation requirements, and advanced weaning features. In cases of severe heart failure, survival was improved by using LVADs when compared to that predicted by solely continuing IABP and drug support. PMID- 15111159 TI - Prolonged 24-hour subzero preservation of heterotopically transplanted rat hearts using antifreeze proteins derived from arctic fish. AB - BACKGROUND: Arctic fish survive subzero temperatures by producing a family of antifreeze proteins (AFPs) that noncolligatively lower the freezing temperature of their body fluids. We report 24-hour storage of mammalian hearts for transplantation at subzero temperatures using AFPs derived from arctic fish. METHODS: Forty-two heterotopic transplantations were performed in isoimmune Sprague-Dawley rats. Harvested hearts were retrogradely infused with cold 4 degrees C University of Wisconsin (UW) solution and were preserved in a specialized cooling bath at two target temperatures, 4 degrees C and -1.3 degrees C for 12,18, and 24 hours (6 experiments/group). Preservation solutions were UW alone for the 4 degrees C group, and UW with 15 mg/mL AFP III for the -1.3 degrees C group. After hypothermic storage the hearts were heterotopically transplanted into isoimmune rats. Viability was assessed and graded on a scale of 0 to 6 (0 = no contractions to 6 = excellent contractions). Transplanted hearts were then fixed in vivo and were subject to electron microscopy and histopathologic examination. RESULTS: None of the hearts preserved at -1.3 degrees C in UW/AFP III solution froze. All control hearts preserved at -1.3 degrees C without AFP protection froze and died at reperfusion. Viability of hearts preserved at -1.3 degrees C in UW/AFP III solution was significantly better after 18 hours of preservation, 30 and 60 minutes after reperfusion (median, 5 versus 3 and 6 versus 3, respectively; p < 0.05) and after 24 hours of preservation 30 and 60 minutes after reperfusion (median, 4.5 versus 1.5 and 5 versus 2, respectively; p < 0.05). Histologic and electron microscopy studies demonstrated better myocyte structure and mitochondrial integrity preservation with UW/AFP III solution. CONCLUSIONS: Antifreeze proteins prevent freezing in subzero cryopreservation of mammalian hearts for transplantation. Subzero preservation prolongs ischemic times and improves posttransplant viability. PMID- 15111160 TI - Hemodilution elevates cerebral blood flow and oxygen metabolism during cardiopulmonary bypass in piglets. AB - BACKGROUND: Hemodilution continues to be widely used during cardiopulmonary bypass (CPB) for both adults and children. Previous studies with nonbypass models have suggested that an increase in cerebral blood flow (CBF) compensates for the reduced oxygen-carrying capacity; however, this increased CBF is achieved by an increase in cardiac output. We hypothesized that even with the fixed-flow perfusion of CPB, CBF would be increased during hemodilution. METHODS: Two experiments were conducted and analyzed separately. In each experiment, 10 piglets were randomized to two different groups, one with a total blood prime yielding a high hematocrit (25% or 30%), and the other with a crystalloid prime resulting in a low hematocrit (10% or 15%). Animals were cooled with pH-stat strategy at full flow (100 or 150 mL.kg(-1).min(-1)) to a nasopharyngeal temperature of 15 degrees C, a period of low flow (50 mL.kg(-1).min(-1)) preceding deep hypothermic circulatory arrest (45 or 60 minutes), and a period of rewarming at full flow. Cerebral blood flow was measured at the beginning of CPB, at the end of cooling, at the end of low flow, 5 minutes after the start of rewarming, and at the end of rewarming by injection of radioactive microspheres. RESULTS: Mean arterial pressure was significantly greater with higher hematocrit at each time point (p< 0.05). Cerebral blood flow and the cerebral metabolic rate of oxygen decreased during cooling and further during low flow bypass but were significantly greater with lower hematocrit during mild hypothermia and at the end of rewarming (p< 0.05). CONCLUSIONS: Hemodilution is associated with decreased perfusion pressure, increased CBF and increased the cerebral metabolic rate of oxygen during hypothermic CPB. PMID- 15111162 TI - Use of a pH-stat strategy during retrograde cerebral perfusion improves cerebral perfusion and tissue oxygenation. AB - BACKGROUND: Although it is well documented that the use of a pH-stat strategy during hypothermic cardiopulmonary bypass improves cerebral blood flow, an alpha stat strategy has been almost exclusively used during retrograde cerebral perfusion. We investigated the effects of pH-stat and alpha-stat management on brain tissue blood flow and oxygenation during retrograde cerebral perfusion in a porcine model to determine if the use of a pH-stat strategy during retrograde cerebral perfusion improves brain tissue perfusion. METHODS: Fourteen pigs were managed by an alpha-stat strategy (alpha-stat group, n = 7) or by a pH-stat strategy (pH-stat group, n = 7) during 120 minutes of hypothermic retrograde cerebral perfusion. Retrograde cerebral perfusion was established through the superior vena cava. Brain tissue blood flow and oxygenation were measured continuously with a laser flowmeter and near infrared spectroscopy, respectively. Brain tissue water content was determined at the end of the experiments. RESULTS: During cooling, brain tissue blood flow was significantly higher with use of the pH-stat strategy than with the alpha-stat strategy (86% +/- 10% versus 40% +/- 3% of baseline). During retrograde cerebral perfusion, brain tissue blood flow was also significantly higher (about three times higher) in the pH-stat group than in the alpha-stat group (15% +/- 4% versus 5% +/- 1% of baseline at 60 minutes of retrograde cerebral perfusion). Tissue oxygen saturation appeared to be higher during retrograde cerebral perfusion in the pH-stat group than in the alpha-stat group. Brain tissue blood flow during rewarming remained significantly higher with the use of pH-stat than with the use of alpha-stat. Brain tissue water contents were similar in both groups. CONCLUSIONS: In our pig model, the use of a pH-stat strategy during retrograde cerebral perfusion significantly improves brain tissue perfusion. Therefore, to improve retrograde cerebral blood flow during retrograde cerebral perfusion, it may be preferable to use a pH-stat strategy, rather than an alpha-stat strategy. PMID- 15111164 TI - Continuous measurement of oxygen consumption during cardiopulmonary bypass: description of the method and in vivo observations. AB - BACKGROUND: Systemic oxygen consumption is not routinely measured during cardiopulmonary bypass, despite its potential benefits. We aimed to develop a noninvasive method to continuously measure oxygen consumption using respiratory mass spectrometry during hypothermic cardiopulmonary bypass in pigs. METHODS: Nine pigs weighing 18.5 (1.6) kg underwent hypothermic (32 degrees C) cardiopulmonary bypass for 180 minutes with 120 minutes of aortic cross clamping. An AMIS 2000 mass spectrometer (Innovision A/S, Odense, Denmark) was adapted for the on-line measurement of oxygen consumption by sampling the inlet and outlet gases of the membrane oxygenator together with measurement of the "expired" gas volume. RESULTS: Active cooling for 60 minutes reduced the venous blood temperature by 2.9 (0.8) degrees C and VO(2) by 0.70 (0.33) mL/kg/min. The 40 minute active rewarming restored the venous blood temperature by 4.4 (0.4) degrees C and oxygen consumption increased by 1.36 (0.33) mL/kg/min. There was wide interanimal variability, however, particularly at higher venous blood temperatures. Immediately after the release of aortic cross clamp, there was a noticeably acute increase in oxygen consumption in all the pigs (0.64 [0.21] mL/kg/min). CONCLUSIONS: A simple and safe adaptation of mass spectrometry allows continuous measurement of oxygen consumption during hypothermic cardiopulmonary bypass. The wide interindividual variations observed in this pilot study underscore the need to more accurately describe changes in oxygen consumption and how they are affected by temperature, oxygen delivery, and other interventions during cardiopulmonary bypass. As such, the technique may have an important role in clinical research and management of oxygen transport in patients undergoing cardiac surgery. PMID- 15111165 TI - A new poly-2-methoxyethylacrylate-coated cardiopulmonary bypass circuit possesses superior platelet preservation and inflammatory suppression efficacy. AB - BACKGROUND: Poly-2-methoxyethylacrylate (PMEA) is a new coating material, and several studies have revealed that PMEA-coated cardiopulmonary bypass (CPB) circuits have good biocompatibility. This study sought to compare this biocompatibility with those of heparin-coated and noncoated circuits. METHODS: Forty-five patients undergoing coronary artery bypass grafting were randomly assigned to PMEA-coated (group P, n = 15), heparin-coated (group H, n = 15), or noncoated (group N, n = 15) circuit groups. Clinical data and the following markers were analyzed: (1) platelet preservation by number of platelets; (2) complement (C) activation by C3a and C4a levels; (3) inflammatory response by interleukin-6 (IL-6) and interleukin-8 (IL-8) levels. RESULTS: Platelet numbers were significantly preserved in group P compared with groups N and H. Postoperative blood loss did not differ among the groups. During CPB, C3a values were significantly lower in group H (536 +/- 145 ng/mL) than in group P (1,458 +/ 433 ng/mL, p < 0.01) and group N (1,815 +/- 845 ng/mL, p < 0.01). The C4a values did not differ 60 minutes after CPB initiation among the groups. The IL-6 and IL 8 levels were significantly lower in group P and group H than in group N. CONCLUSIONS: The PMEA coating was superior to heparin coating and noncoating in preserving platelets, and was equivalent to heparin coating in terms of the perioperative clinical course and inhibition of inflammatory cytokines, but slightly inferior in reducing complement activation. PMID- 15111166 TI - Myocyte contractility with caspase inhibition and simulated hyperkalemic cardioplegic arrest. AB - BACKGROUND: Exposure of left ventricular (LV) myocytes to simulated hyperkalemic cardioplegic arrest (HCA) has been demonstrated to perturb ionic homeostasis and adversely affect myocyte contractility on rewarming. Altered ionic homeostasis can cause cytosolic activation of the caspases. While caspases participate in apoptosis, these proteases can degrade myocyte contractile proteins, and thereby alter myocyte contractility. Accordingly, this study tested the hypothesis that caspase inhibition during HCA would attenuate the degree of myocyte contractile dysfunction upon rewarming, independent of a loss in myocyte viability. METHODS: Porcine (n = 8) LV myocytes were isolated and assigned to the following treatment groups: normothermic control: incubation in cell culture media for 2 hours at 37 degrees C; HCA only: incubation for 2 hours in hypothermic HCA solution (4 degrees C, 24 mEq K(+)); or incubation in hypothermic HCA solution supplemented with 10 microM of the caspase inhibitor, z-VAD (z-Val-Ala-Asp-fluoromethyl ketone, HCA+zVAD). Myocyte viability, assayed as a function of mitochondrial function, was determined to be similar in the normothermic and both HCA groups. RESULTS: The HCA caused a significant reduction in myocyte shortening velocity compared with normothermic control values (41 +/- 6 versus 86 +/- 8 microm/s, p < 0.05). The HCA+zVAD group had significantly improved myocyte shortening velocity compared with the HCA only group (63 +/- 7 microm/s, p < 0.05). CONCLUSIONS: Independent of changes in viability, caspase inhibition attenuated myocyte contractile dysfunction after HCA and rewarming. Thus, caspase activation during HCA contributes, at least in part, to impaired myocyte contractility with rewarming. Supplementation of HCA with caspase inhibitors may provide a means to preserve myocyte contractile function after cardioplegic arrest. PMID- 15111168 TI - Selective timing for the arterial switch operation. AB - BACKGROUND: To determine outcomes for the arterial switch operation individualized according to the underlying anatomy and clinical status. METHODS: A retrospective review of a consecutive series of infants less than 90 days of age who underwent the arterial switch operation at a single institution. RESULTS: From July 1993-April 2001, 117 infants underwent an arterial switch operation before 90 days of age. Seventy-five patients (64%) had transposition of the great arteries with intact ventricular septum with the aim of operation before 14 days of age; however, 8 of these patients had delayed presentation (range 15-46 days). Thirty-five patients (30%) had transposition with a ventricular septal defect (30 patients) or double outlet right ventricle (5 patients) and normal arch anatomy and were repaired within the first 90 days of life depending on the severity of heart failure at a median of 12 days of age (range 3-83 days). Seven patients (6%) had associated aortic coarctation (5 patients) or interrupted aortic arch (2 patients). One patient died during hospitalization (0.85% hospital mortality) and one patient died from noncardiac causes during a median follow-up of 35 months (1.7% total mortality). Four patients required intervention during follow-up (3.4%) for new aortic coarctation (2 patients), supravalvar pulmonic stenosis (1 patient), or right hemi-diaphragm paralysis (1 patient). CONCLUSIONS: Individualized timing for the arterial switch operation within the first ninety days of life produces excellent survival rates for all types of transposition physiology with the expectation of a satisfactory course during follow-up. PMID- 15111170 TI - Preoperative brain injury in newborns with transposition of the great arteries. AB - BACKGROUND: The objective was to determine the timing and mechanism of brain injury using preoperative and postoperative magnetic resonance imaging (MRI) and three-dimensional MR spectroscopic imaging (MRSI) in newborns with transposition of the great arteries (TGA) repaired with full-flow cardiopulmonary bypass. METHODS: Ten term newborns with TGA undergoing an arterial switch operation were studied with MRI, MRSI, and neurologic examination preoperatively and postoperatively at a median of 5 days (2 to 9 days) and 19 days (14 to 26 days) of age, respectively. Five term historical controls were studied at a median of 4 days (3 to 9 days). Lactate/choline (marker of cerebral oxidative metabolism) and N-acetylaspartate (NAA)/choline (marker of cerebral metabolism and density) were measured bilaterally from the basal ganglia, thalamus, and corticospinal tracts. RESULTS: Four TGA newborns had brain injury on the preoperative MRI. The only new lesion detected on the postoperative study was a focal white matter lesion in one newborn with a normal preoperative MRI. The MRSI of age-adjusted lactate/choline was quantitatively higher in newborns with TGA compared with those without heart disease (p < 0.0001), even in newborns without MRI evidence of preoperative brain injury. Lactate/choline decreased after surgery but remained elevated compared with controls. In newborns with TGA, those with preoperative brain injury on MRI had lower NAA/choline globally (p = 0.04) than those with normal preoperative MRI. Five newborns had a decline in NAA/choline from the preoperative to postoperative studies. CONCLUSIONS: Abnormal brain metabolism and injury was observed preoperatively in newborns with TGA. Brain injury is not solely related to the operative course. PMID- 15111171 TI - Contegra bovine jugular vein right ventricle to pulmonary artery conduit in Ross procedure. AB - BACKGROUND: In the Ross procedure a valved conduit, most commonly pulmonary or aortic homograft, is used in place of autotransplanted pulmonary valve. Increasing demand and diminishing supply of homografts has resulted in a search for alternatives. A biological conduit from the valved segment of bovine jugular vein (Contegra) has been used successfully as an alternative. METHODS: Early clinical and echocardiographic results were analyzed retrospectively for 20 patients (median age 14.4 years) who underwent a Ross procedure with Contegra as right ventricle to pulmonary artery conduit between November and June 2003 (during the last 31 months). RESULTS: There was no operative mortality and late mortality or morbidity during the mean follow-up of 13.8 +/- 9.1 months (range 1 to 31 months). No patient required reoperation. The median gradient at discharge was 16 +/- 4.5 mm Hg, which remained unchanged at last follow-up. No deterioration in conduit or conduit valve function was noted. CONCLUSIONS: This new bovine jugular vein conduit can be a viable alternative to a homograft in the Ross procedure. The early clinical and hemodynamic results are encouraging. Ease of availability and favorable handling and technical characteristics make it more attractive than a homograft. Xenograft origin of this conduit necessitates close follow-up for assessment of durability and longer-term results. PMID- 15111172 TI - Medtronic Freestyle valve for right ventricular reconstruction in pediatric Ross operations. AB - BACKGROUND: The use of homograft conduits to reconstruct right ventricle (RV) to pulmonary artery (PA) connections is an essential component of the Ross operation. Homograft availability and cost may be problematic when considering the Ross operation. We elected in January 1998 to utilize commercially available xenografts as an alternative to homografts for RV/PA reconstruction in the pediatric Ross operation. Our early results using the Medtronic Freestyle valve (Medtronic, Minneapolis, MN) for RV/PA reconstruction are presented. METHODS: We reviewed our database for all Ross operations performed on children since January 1998. A total of 16 patients were identified. Eleven children received a Medtronic Freestyle valve, 2 children received a homograft, and 3 children received another type of xenograft. Echocardiographic evaluation of all children who received the Medtronic Freestyle valve was performed at hospital discharge and at two subsequent outpatient evaluations. RESULTS: The median peak instantaneous pressure gradient across the xenograft was 16 +/- 9 mm Hg (immediately after surgery before hospital discharge); 22 +/- 20 mm Hg at 23 +/- 11 months (first postdischarge follow-up); and 27 +/- 20 mm Hg at 35 +/- 9 months (second postdischarge follow-up). Linear regression analysis revealed an increasing pressure gradient with time (R(2)-adjusted = 0.44, p < 0.0001). At the same three observation points, the xenograft annulus diameter decreased: 25 +/- 1.2 mm; 19 +/- 4.3 mm; and 20 +/- 1.8 mm. Linear regression analysis revealed a decreasing annulus diameter with time (R(2)-adjusted = 0.41, p < 0.0001). CONCLUSIONS: The Medtronic Freestyle valve provides a possible alternative to homografts for the reconstruction of the RV/PA connection in the pediatric Ross operation. Long-term follow-up is necessary to evaluate this xenograft as an alternative to the homograft. PMID- 15111173 TI - Repair of complete atrioventricular septal defects in patients weighing less than 5 kg. AB - BACKGROUND: The aim of this study was to evaluate the impact of weight less than 5 kg at operation on mortality and morbidity in patients with atrioventricular septal defect (AVSDc) undergoing total correction. METHODS: Between January 1990 and December 2002, 190 consecutive patients with AVSDc underwent total biventricular correction. They were divided into two groups: group I (n = 64 patients weighing < 5 kg) and group II (n = 126 patients weighing > 5 kg). Associated major cardiac malformations were found in 49 (25.8%) patients. Associated left atrioventricular valve (LAVV) malformations were found in 35 (18.4%) patients. The mean follow-up time was 4.1 +/- 2.9 years (range 2 months 10.7 years). RESULTS: The in-hospital mortality in group I was 7.8% (5 patients) versus 8.7% (11 patients) in group II (p = 0.95). Major associated cardiac malformations (p < 0.001) and pulmonary hypertension (p = 0.006) were found to be strong predictors for poor postoperative survival. At discharge the mean LAVVR grade in group I was 1.45 +/- 1.2 versus 1.2 +/- 1 in group II (p = 0.13). The actuarial overall survival rates at 1, 3, 5, and 7 years were 96.5%, 92.5%, 91.5%, and 89% respectively and the actuarial overall reoperation free survival rates at 1, 3, 5, and 7 years were 95%, 87%, 84%, and 73%. Twenty-three patients underwent reoperation due to severe left atrioventricular valve regurgitation (LAVVR). Strong predictors for overall reoperation free survival were the operation year before 1995 (p < 0.001), postoperative LAVVR greater than or equal to 2 (p = 0.006), major associated cardiac malformations (p = 0.00034), associated LAVV malformations (p = 0.0044), and non or partial LAVV cleft closure (p = 0.012). The actuarial survival rates between patients weighing less than 5 kg versus patients weighing more than 5 kg were similar (p = 0.51); instead the overall reoperation free survival was significantly lower in patients weighing less than 5 kg (p = 0.022) according to the log-rank test. Weight less than 5 kg (p = 0.023, beta = -0.6) was one of the predictors for reoperation due to severe LAVVR in this series. CONCLUSIONS: We may conclude that in the current era repair of AVSDc can be carried out successfully in patients less than 5 kg, however, weight less than 5 kg at initial complete repair seems to be a predictor for late reoperation due to LAVVR. Suture separation at the cleft site or between the leaflets of the newly created mitral valve and the patch remain the main causes of postoperative LAVVR in patients weighing less than 5 kg. PMID- 15111174 TI - Impact of right ventricle to pulmonary artery conduit on outcome of the modified Norwood procedure. AB - BACKGROUND: To determine and compare outcome of the modified Norwood procedure using either a systemic to pulmonary artery (SPA) shunt or right ventricle to pulmonary artery (RV-PA) conduit in a consecutive series of neonates at a single institution. METHODS: The medical records were retrospectively examined for preoperative demographic and echocardiographic data, operative variables, and postoperative clinical and hemodynamic data. From November 2001 to March 2003, 21 neonates had a modified Norwood procedure (SPA shunt, n = 8; RV-PA conduit, n = 13) at a median age of 5 days (range 1 to 18 days) and a median weight of 2.9 kg (range 1.7 to 4.1 kg). Of the 21 infants, 12 were considered high risk due to presence of low birth weight (n = 4), extracardiac or genetic anomalies (n = 5) or obstruction to pulmonary venous return (n = 5). Nine "high risk" infants were in the RV-PA conduit group. RESULTS: Overall Norwood operation survival was 90% (19/21) and did not differ between groups. There were 2/19 interstage deaths and Kaplan-Meier survival at 1 year is 79%. Neonates in the RV-PA conduit group had significantly higher diastolic blood pressures at 1, 6, and 24 hours postoperatively (p < 0.05). Neonates in the SPA shunt group had significantly higher heart rates at 1 hour postoperatively (p < 0.05) than those in the RV-PA group. There was a trend to higher number of ventilatory interventions to balance Qp:Qs in the SPA shunt group (p = 0.06). CONCLUSIONS: In a relatively high-risk group, neonates having an RV-PA conduit as part of the Norwood procedure have favorable postoperative hemodynamics and a good likelihood of stage I survival. PMID- 15111175 TI - Mycophenolic mofetil reduces the HLA antibody response of children to valved allograft implantation. AB - BACKGROUND: Valved allografts induce a brisk, broadly reactive human leukocyte antigen (HLA) antibody response in children after implantation. Mycophenolic mofetil (MMF) is a powerful immunosuppressant that inhibits the proliferation of both T cells and B cells and has been reported to possibly reduce HLA panel reactive antibody (PRA) in sensitized transplant recipients. METHODS: The purpose of this study was to determine whether MMF can blunt the HLA antibody response to valved allografts in children. Eight patients completed (of 28 approached) a pilot study to determine the effects of 3 months of twice daily MMF (600 mg/m(2)/dose) on the HLA antibody response measured before surgery, at 1 month, and at 3 months after implantation. Patients were 7.5 +/- 4 yrs old (mean +/- standard deviation [SD]), with 5 patients undergoing repair of tetralogy of Fallot, 2 Ross procedures, and 1 aortic valve replacement. RESULTS: In contrast to historical controls with a virtual 100% HLA class I PRA response to valved allograft implantation, MMF markedly decreased the HLA class I antibody response at 1 and 3 months postimplantation. In 6 cases where the HLA type of the donor was defined, PRA specificity correlated with incompatible antigens on the allograft. One patient withdrew after 2 weeks due to a sinus infection that was successfully treated with oral antibiotics, and 3 patients had a transient adverse effect of postoperative vomiting. CONCLUSIONS: This study demonstrates the ability to pharmacologically abrogate the HLA class I antibody response to valved allograft implantation in children using MMF. PMID- 15111177 TI - Long-term results of operation for 420 patients with early squamous cell esophageal carcinoma discovered by screening. AB - BACKGROUND: Cancer of the esophagus is one of the most commonly seen malignancies in China. From 1959 to 1981, mass screening of esophageal cancer disclosed that the age-adjusted incidence in the 40- to 69-year-old population in Lin County, Henan Province, was 470/10(5) In its northern part, an even higher incidence of 760/10(5)was found. As they were discovered by mass screening, most of them were found to have early lesions. Surgical treatment was done in attempt to find out the feasibility of managing esophageal carcinoma by early diagnosis and early treatment. This paper is the result of the long-term follow-up. METHODS: Since 1972, a total of 17 extensive mass screening has been conducted among more than 160,000 participants in the rural areas in Henan, Hebei, and northern Jiangsu provinces, sorting out more than 30,000 high-risk individuals. Among these individuals, 24,600 were examined by endoscopy, discovering 2,094 patients with carcinomas in the esophagus or gastric cardia; 757 of these 2,094 patients were found to have superficial esophageal cancer; 420 patients accepted surgical treatment. Esophagectomy with gastric replacement was performed through left thoracotomy in all patients. Cervical anastomosis 94 (22.4%), intrathoracic supraaortic anastomosis 307 (73.1%), and infra-aortic anastomosis 19 (4.5%) were done. Double thoracoabdominal lymphatic dissection was performed. RESULTS: The resection rate was 100%. One-month operative mortality occurred in 5 (1.2%). Postoperative complications developed and were satisfactorily treated in 28 patients (6.7%). Pathology of the cancer specimens showed that there were carcinoma in situ in 76 (all without lymphatic metastasis), intramucosal (TI) carcinoma in 126 (2 [1.6%] with lymphatic metastasis), and submucous infiltrating (TI) carcinoma in 218 (34 [15.6%] with lymphatic metastasis). All these 420 patients have been followed up to 2001 with a follow-up rate of 94.1%. Those who were lost to follow-up were taken as censored cases. The survival rates were calculated by the life-table method. The 5-, 10-, 15-, 20-, and 25-year survival rates were 86.14%, 75.03%, 64.48%, 56.17%, and 49.93%, respectively. CONCLUSIONS: Esophageal balloon cytology, endoscopy, mucosa 1.2% iodine stain, and multipoint biopsy may be the best approach for early diagnosis of esophageal carcinoma. Surgical resection of superficial esophageal cancer provides excellent long-term survival with acceptable quality of life. It was discovered that carcinoma in situ and intramucosal carcinoma gave far better results than the submucosal infiltrative carcinoma, as the latter tends to have a higher frequency of lymphatic metastasis. PMID- 15111178 TI - Experience with programmed steroid treatment with thymectomy in nonthymomatous myasthenia gravis. AB - BACKGROUND: The benefit of thymectomy in myasthenia gravis management is recognized but the perioperative course can fluctuate. The goal of this study was to assess the feasibility and clinical benefit of dose-escalated steroid therapy with thymectomy for nonthymomatous myasthenia gravis. METHODS: We reviewed the records of 69 myasthenia gravis patients who were followed up after undergoing transsternal thymectomy with extended anterior mediastinal dissection in our hospital between 1976-2000. Forty-eight patients in the programmed treatment group who had dose-escalated and de-escalated steroid therapy during the perioperative period comprised 17 patients with ocular myasthenia gravis and 31 patients with generalized myasthenia gravis. Clinical benefits and clinical remission, which was diagnosed when the patients were symptom-free without medications for at least 1 year, were compared with those of 21 patients in the occasional treatment group who received medications occasionally over the perioperative period. RESULTS: Postoperative respiratory failure and myasthenic crisis did not occur in the programmed treatment group but did occur in 6 patients in the occasional treatment group. Remission rates in the programmed treatment group (mean follow-up, 6.4 years) were 30% at 3 years, 38% at 5 years, and 46% at 10 years; rates in the occasional treatment group (mean follow-up, 9.6 years) were 25% at 3 years, 25% at 5 years, and 45% at 10 years. CONCLUSIONS: Programmed steroid therapy in patients with nonthymomatous myasthenia gravis is feasible and it provides clinical benefit when fluctuating symptoms occur during the perioperative period. PMID- 15111179 TI - Upregulated hypoxia-inducible factor-1 DNA binding activity to the vascular endothelial growth factor-A promoter mediates increased vascular permeability in donor lung grafts. AB - BACKGROUND: Transplantation-induced hypoxia results in enhanced vascular permeability and tissue vascular endothelial growth factor (VEGF) and endothelin 1 (ET-1) overexpression in donor lung grafts. Promoter studies have uncovered a hypoxia-inducible factor (HIF)-1 binding site (HBS) in 5'-flanking region of VEGF gene that regulates the hypoxia-induced expression of VEGF; and ET-1 potently stimulates VEGF-A production. We hypothesized that HIF-1 regulates VEGF-mediated vascular permeability in lung grafts. METHODS: We studied the mRNA and protein expression of HIF-1 and its protein-binding capacity to the HBS of the VEGF gene in biopsies of preserved donor and control lungs, using real-time reverse transcription-polymerase chain reaction, Western blotting, and electrophoretic mobility shift assay. Wet-to-dry lung weight ratio was measured in donor and control lungs. RESULTS: While HIF-1 alpha mRNA expression was unchanged, HIF-1 beta was downregulated (p < 0.05) in donor versus control lungs. Protein expression of both, HIF-1 alpha and -beta was significantly upregulated in donor lung grafts. HIF-1 binding to the HBS of the VEGF promoter as well as tissue fluid content were increased in donor lung biopsies versus controls (p < 0.05). CONCLUSIONS: These data indicate that upregulated HIF-1 DNA binding activity to the HBS of VEGF-A most likely contributes to elevated VEGF levels in preserved lung grafts. Unchanged HIF-1 alpha mRNA expression did not affect HIF-1 alpha protein levels. Endothelin-1 increases HIF-1 alpha accumulation and activates HIF 1 transcription complex in vitro. Therefore, ET-1-mediated increased HIF-1 alpha protein stability most likely leads to transcriptional activation of VEGF during lung graft preservation. Targeting HIF might be of benefit to counteract edema formation in preserved lung grafts. PMID- 15111180 TI - A novel technique for localization and excisional biopsy of small or ill-defined pulmonary lesions. AB - BACKGROUND: The purpose of this study was to develop and evaluate radiotracer guided localization of small or ill-defined pulmonary nodules for thoracoscopic excisional biopsy. METHODS: This study consisted of two parts: a laboratory study in rats to determine the most suitable radiotracer, and a pilot study in humans to determine the feasibility of radiotracer lung nodule localization. The right lung of 12 rats was injected with a technetium 99m (Tc 99m) based radiotracer solution: 4 each with macroaggregated albumin (MAA), unfiltered sulfur colloid (SC), and pertechnetate (TcO(4)). Serial imaging was performed using a small animal gamma camera for 4 hours following injection. In 13 patients, computed tomographic (CT) guided injection of Tc 99m MAA solution was made into or adjacent to a pulmonary nodule suspicious for primary lung cancer. Gamma probe localization of the nodule was performed during subsequent thoracoscopic surgery. RESULTS: In the animal model, MAA provided more precise localization than SC or TcO(4) and was selected for the human study. In the human series, all 13 patients had successful gamma probe localization of their lesion. There were no radiologic or surgical complications. Seven of 13 nodules were malignant, and five of these nodules were stage IA primary lung carcinomas. CONCLUSIONS: Computed tomographic guided radiotracer localization of small or ill-defined pulmonary nodules using Tc 99 m MAA before thoracoscopic excisional biopsy is feasible and may make excisional biopsy the preferred management strategy for the management of small pulmonary nodules in patients at high risk for malignancy. PMID- 15111182 TI - Endoscopic ultrasound in lung cancer patients with a normal mediastinum on computed tomography. AB - BACKGROUND: Computed tomography (CT) is the most common method of staging lung cancer. We have previously shown endoscopic ultrasound guided fine-needle aspiration (EUS-FNA) to be highly accurate in staging patients with nonsmall cell lung cancer (NSCLC) who have enlarged mediastinal lymph nodes on CT scan. In this study we report the accuracy and yield of EUS-FNA in staging patients without enlarged mediastinal lymph nodes by CT. METHODS: Patients with NSCLC and CT scan showing no enlarged mediastinal lymph nodes (> 1 cm for all nodes except > 1.2 cm for subcarinal) in the mediastinum underwent EUS. Fine needle aspiration was performed on at least one lymph node, if present, in the upper mediastinum, aortopulmonary window, subcarinal, and periesophagus regions. Each specimen was evaluated with on-site cytopathology and confirmed with complete cytopathologic examination. RESULTS: Sixty-nine patients without enlarged mediastinal lymph nodes were evaluated. Endoscopic ultrasound detected malignant mediastinal lymph nodes in 14 of 69 patients as well as other advanced (American Joint Committee on Cancer [AJCC] stage III/IV) in 3 others (1 left adrenal, and 2 with mediastinal invasion of tumor) for a total of 17 of 69 (25%, 95% confidence interval: 16% to 34%) patients. Eleven additional patients were found to have advanced disease by bronchoscopy (2), mediastinoscopy (2), and thoracotomy with mediastinal lymph node dissection (7). The sensitivity of EUS for advanced mediastinal disease was 61% (49% to 75%), and the specificity was 98% (95% to 100%). CONCLUSIONS: Endoscopic ultrasound guided fine needle aspiration can detect advanced mediastinal disease and avoid unnecessary surgical exploration in almost one of four patients who have no evidence of mediastinal disease on CT scan. In addition to previously reported results in patients with enlarged lymph nodes on CT, these data suggest that all potentially operable patients with nonmetastatic NSCLC may benefit from EUS staging. PMID- 15111183 TI - Visceral pleural involvement in nonsmall cell lung cancer: prognostic significance. AB - BACKGROUND: A tumor of any size that invades the visceral pleura is classified in the T2 category; however, the definition of the visceral pleural involvement has remained somewhat ambiguous. It is unclear whether the T2 category includes the p2 status alone or incorporates the extent of the p1 status. METHODS: We retrospectively analyzed the survival of 474 patients with T1 and T2 nonsmall cell lung cancer to evaluate the influence of the degree of visceral pleural involvement (p0, p1, and p2) on the prognosis and to clarify the definition of the visceral pleural involvement. RESULTS: The 5-year survival rates according to the degree of visceral pleural involvement were 68.0% in p0 (n = 345), 43.9% in p1 (n = 110), and 54.9% in p2 (n = 19; p0 versus p1, p = 0.0004; p0 versus p2, p = 0.013; and p1 versus p2, p = 0.61). The degree of visceral pleural involvement (p0 versus p1/p2) was a significant independent prognostic factor from tumor size and lymph node involvement, by multivariate analysis (relative risk = 1.47, p = 0.033). The prognosis of pN0 patients with p1 and tumor size 3 cm or less was significantly poorer than that of those with p0 and tumor size 3 cm or less (p = 0.0004), and the prognosis of patients with p1 and tumor size more than 3 cm was significantly poorer than that of those with p0 and tumor size more than 3 cm (p = 0.024). CONCLUSIONS: The degree of visceral pleural involvement (p0 versus p1/p2) is an important component of the lung cancer staging system. Tumors with p1 and p2 status should be regarded as representing visceral pleural involvement and T2 disease. PMID- 15111185 TI - Accuracy of virtual bronchoscopy to detect endobronchial lesions. AB - BACKGROUND: Virtual bronchoscopy (VB) could obviate flexible bronchoscopy (FB) if no endobronchial lesion is detected in patients presenting with a suspicion of malignancy. Our objectives were to evaluate the accuracy (in terms of sensitivity and specificity) of VB in detecting endobronchial lesions, and to determine the anatomical limit of detection of endobronchial lesions by VB. METHODS: This study involved, in a blind comparison of VB and FB, consecutive patients presenting with symptoms or plain chest radiography abnormalities raising the suspicion of pulmonary neoplasm. After the standard chest computed tomography (CT), additional helical CT data were acquired from the aortic arch to the origin of the segmental bronchi of the inferior lobes in one 20-second breath hold using an helicoidal CT scan (3.0-mm collimation with a pitch of 1.5 and 1.5-mm reconstruction intervals). RESULTS: One hundred ninety patients were enrolled; 136 patients (including 63 with an endobronchial lesion at FB) contributed to the primary analysis. The sensitivity and specificity of VB to detect endobronchial lesions were 68% (95% confidence interval [CI]: 55% to 79%) and 90% (95% CI: 81% to 96%), respectively. Overall, the agreement between VB and FB regarding the location on endobronchial lesions was substantial (weighted kappa: 0.66). However, VB detected only 26 of the 34 lobar lesions (sensitivity: 76%; CI: 59% to 89%) and 11 of the 23 segmental lesions (sensitivity: 48%; CI: 27% to 69%). CONCLUSIONS: Beyond the mainstem bronchi, VB is not accurate enough to detect endobronchial lesions and to obviate FB in patients presenting with a suspicion of malignancy. PMID- 15111186 TI - Bronchial carcinoid tumors: nodal status and long-term survival after resection. AB - BACKGROUND: Bronchial carcinoid tumors show a favorable outcome. We sought to determinate the variables influencing the long-term survival of patients treated for bronchial carcinoid tumors. METHODS: We conducted a retrospective single institutional review of 163 patients surgically treated from January 1990 to April 2002. According to 1999 World Health Organization criteria, cases were segregated into typical (<2 mitoses per 2 mm, no necrosis) and atypical carcinoids (2 to 10 mitoses per 2 mm or necrosis). RESULTS: There were 86 men and 77 women with a mean age of 49.5 +/- 11 years. Symptoms were present in 89 patients (54.6%). Operations included 145 formal lung resections (89%), 9 wedge resections (5.52%), 8 sleeve lobectomies (4.9%), and 1 segmental resection (0.61%) plus radical mediastinal lymphadenectomy in all cases. No operative mortality was reported. Histologic examination showed 121 (74.2%) typical carcinoids (107 N0 and 14 N1), and 42 (25.8%) atypical carcinoids (15 N0, 18 N1, 9 N2). All patients were included in a follow-up (median, 54 months; mean, 58 months; range, 4 to 150 months), which included total body computed tomographic scan and bronchoscopy every year. Overall 5-year survival was 90.3% with a mean survival time of 139 months (95% confidence interval, 133 to 145). In N0 patients with either typical or atypical carcinoid tumors, no disease-related mortality was reported (100% 5-year survival). In N1 patients, 5-year survival was 90.0% for those with typical carcinoids, and 78.8% for those with atypical carcinoids (p = 0.394). In atypical carcinoids with N2 disease, 5-year survival was 22.2%. CONCLUSIONS: Prognosis in bronchial carcinoid tumors is more related to nodal status than to histologic subtype. In N0 and N1 patients no statistical significant difference has been found between typical and atypical subtype. However, N2 bronchial carcinoid tumors show a dismal prognosis. PMID- 15111188 TI - Prospective randomized study evaluating a biodegradable polymeric sealant for sealing intraoperative air leaks that occur during pulmonary resection. AB - BACKGROUND: To evaluate the safety and effectiveness of a new biodegradable polymeric sealant to close intraoperative air leaks after pulmonary resection. METHODS: In a multicenter prospective randomized trial, 161 patients with a median age of 67 years old (range 18-85 years old), were randomized in a 2:1 ratio to receive sealant or control for at least one significant air leak (> or = 2.0 mm in size) after pulmonary resection. In the sealant group, all significant air leaks underwent attempted repair by standard methods (sutures, staples, or cautery) prior to the application of sealant. The control group underwent only standard methods. Blood was analyzed for immunologic response. Patients were followed up 1 month after surgery. RESULTS: Intraoperative air leaks were sealed in 77% of the sealant group compared with 16% in the control group (p < 0.001). The sealant group had significantly fewer patients with postoperative air leaks compared with the control group (65% vs 86%, p = 0.005). Median length of hospitalization was 6 days (range, 3-23 days) for the sealant group compared with 7 days (range 4-38 days) for controls (p = 0.028). There was no difference in mortality, morbidity, duration of chest tubes, or immune responses between the two groups. CONCLUSIONS: This study demonstrates the effectiveness of a biodegradable polymer when used as an adjunct to standard closure methods for sealing significant intraoperative air leaks that develop from pulmonary surgery. Use of the sealant led to a reduction in postoperative air leaks, which may have decreased the length of hospitalization. PMID- 15111187 TI - Prognostic significance of lymph node metastasis found during pulmonary metastasectomy for extrapulmonary carcinoma. AB - BACKGROUND: The prognostic significance of lymph node metastasis in cancer patients is well documented. Pulmonary metastasectomy in selected patients is associated with improved survival. Little is known about the prognostic significance of lymph node metastases found during pulmonary metastasectomy for extrapulmonary carcinoma metastatic to the lung. METHODS: The records of all patients who underwent pulmonary metastasectomy and complete mediastinal lymph node dissection for extrapulmonary carcinomas at our institution from November 1985 through July 1999 were reviewed. RESULTS: Eight hundred eighty-three patients underwent pulmonary metastasectomy. Of these, 70 patients (7.9%) (44 men, 26 women) had concomitant complete lymphadenectomy. Median age was 64 years (range, 33 to 83 years). Median time interval between primary tumor resection and metastasectomy was 34 months (range, 0 to 188 months). Wedge excision was performed in 46 patients, lobectomy in 16, both in 7, and pneumonectomy in 1. Lymph node metastases were found in 20 patients (28.6%) and were classified as intrapulmonary or hilar (N1) in 9, mediastinal (N2) in 8, and both in 3. There were no operative deaths. Median follow-up was 6.6 years (range, 1.1 to 14.6 years). Three-year survival for patients with negative lymph nodes was 69% as compared with only 38% for those with positive lymph nodes (p < 0.001). CONCLUSIONS: The presence of lymph node metastases at the time of pulmonary metastasectomy for extrapulmonary carcinoma has an adverse effect on prognosis. Complete mediastinal lymph node dissection should be considered at the time of pulmonary metastasectomy for carcinoma to improve staging and guide treatment. PMID- 15111189 TI - Lung wedge resection improves outcome in stage I primary spontaneous pneumothorax. AB - BACKGROUND: To evaluate the role of apical lung wedge resection in patients with recurrent primary spontaneous pneumothorax with no endoscopic abnormalities at surgery as compared with simple apical pleurectomy. METHODS: We performed a retrospective analysis on 126 consecutive video-assisted thoracoscopic surgery (VATS) procedures in 113 patients treated for stage I recurrent PSP between January 1994 and December 2001. Two surgical strategies were applied: simple apical pleurectomy (57 procedures, 45.2%: group A) and apical pleurectomy together with an apical lung wedge resection (69 procedures, 54.8%: group B). RESULTS: Mean duration of chest tubes was 1.4 days (range, 1 to 7), mean hospital stay was 2.4 days. Three patients (2.4%) required redo VATS, 2 in group A (3.5%) for persistent air leak and 1 (1.4%) in group B for apical hematothorax. Mean follow-up was 38.7 months. Overall recurrence rate was 3.2%. Four patients in group A (7%) experienced recurrent ipsilateral pneumothoraces 4 to 73 weeks (mean, 30.2) after surgery. No recurrences were observed in group B (p = 0.009). CONCLUSIONS: In this selected group of patients without endoscopical abnormalities, VATS offers low recurrence rates. However, these data suggest that apical pleurectomy should be accompanied by apical lung wedge resection even for this favorable category of patients. PMID- 15111190 TI - Heart transplantation in a patient with isolated noncompaction of the left ventricular myocardium. AB - We describe a patient with isolated noncompaction of the left ventricle who presented with worsening congestive heart failure and was successfully treated with heart transplantation. The prognosis for these patients is poor because of accelerated event rates of fatal arrhythmias, thromboemboli, and profound left ventricular decompensation. Only 7 patients with isolated noncompaction of the left ventricle have been reported to have undergone heart transplantation. Herein we describe a patient with isolated noncompaction of the left ventricle who underwent successful heart transplantation. PMID- 15111191 TI - Monitoring weaning from BIVAD Thoratec with peak oxygen consumption. AB - A biventricular assistance device has been implanted in a young woman for a peripartum cardiac failure. An intended weaning consisted of gradual reloading and exercise training monitored with peak oxygen consumption (VO(2)) and radionuclide-left ventricle ejection fraction. Progressive increase in peak VO(2) during partial assistance occurred more than 2 months, from 10.3 to 19 mL.kg( 1).min(-1). Successful explantation was realized when peak VO(2) exceeded 15 mL.kg(-1).min(-1) and radionuclide-left ventricle ejection fraction was more than 40% during off-pump testing. PMID- 15111192 TI - Mitral valve repair in a 55-year-old man with left lung agenesis. AB - We report on a 55-year-old man who was diagnosed with agenesis of the left lung in childhood. He was essentially asymptomatic until he was 53 years of age, when he became symptomatic with exertional dyspnea due to severe mitral regurgitation. We performed mitral valve repair using a median sternotomy incision approach. PMID- 15111193 TI - Transesophageal echocardiography-guided chordal replacement for tricuspid regurgitation. AB - A 29-year-old man was referred to our institution for severe tricuspid regurgitation. Preoperative transesophageal echocardiography revealed dilation of the tricuspid annulus with prolapse of the anterior leaflet and progressive increase in right ventricular diameter. The anterior papillary muscle was elongated. Annuloplasty was completed by polytetrafluoroethylene artificial chordae implanted on the free margin of the anterior leaflet, and thereafter, tied outside the right ventricle, under echocardiography control to achieve appropriate length. The patient recovered well, and 6 months after surgery, repeat echocardiography revealed only mild tricuspid regurgitation with recovery of normal right ventricular dimensions. PMID- 15111194 TI - Delayed rupture of a postinfarction left ventricular true aneurysm. AB - Delayed rupture of a true left ventricular aneurysm is a rare clinical condition. We report a case of a 60-year-old woman who underwent emergency surgical repair of a ruptured true aneurysm of the left ventricular inferior wall 3 months after the myocardial infarction. The repair consisted of endoaneurysmorraphy patch technique. The patient made a satisfactory recovery. PMID- 15111195 TI - A man with saphenous vein graft aneurysms after bypass surgery. AB - Saphenous vein graft (SVG) aneurysms are a rare complication of coronary artery bypass graft surgery. Patients in whom these aneurysms form a fistula with either a cardiac chamber or mediastinal vessel are even more uncommon and present a difficult diagnostic and therapeutic challenge. We present a patient with SVG aneurysms and a fistula to the left atrium. PMID- 15111196 TI - Fatal complications after use of the Symmetry Aortic Connector in coronary artery bypass surgery. AB - During the last 2 years, 103 aortic saphenous vein graft anastomoses were performed in 68 patients undergoing off-pump coronary artery bypass by using the Symmetry Bypass System Aortic Connector. Of these patients, 2 died during the early postoperative period. In the first patient, after an episode of ventricular fibrillation and closed-chest cardiac massage, the sternum was opened and hemopericardium secondary to leakage of the proximal anastomotic device was found. The second patient died of ascending aortic dissection, the tear of which was likely to have originated from the proximal anastomotic site. PMID- 15111197 TI - Coronary artery bypass grafting in an immune thrombocytopenic purpura patient using off-pump techniques. AB - We performed an off-pump coronary artery bypass grafting (OPCABG) procedure on a 60-year-old woman with idiopathic thrombocytopenic purpura (ITP) whose platelet count was 42 x 10(3) per microliter on admission. She was treated with immunoglobulin G (IgG) (0.5 g.kg(-1).d(-1)) for 4 days, resulting in a platelet count rise to 187 x 10(3) per microliter. She subsequently underwent an uneventful OPCABG procedure without requiring any blood transfusions. The combination of OPCABG and preoperative IgG therapy appears to be an ideal strategy for ITP patients requiring coronary revascularization. PMID- 15111198 TI - Surgical treatment of five patients with aortobronchial fistula in the aortic arch. AB - Aortobronchial fistula (ABF) is a rare condition that is almost always fatal in the absence of prompt and proper treatment. However, treatment remains challenging, particularly in the aortic arch. We present six operations for 5 such patients, in which no in-hospital deaths occurred. One patient with mycotic aneurysm died suddenly 10 months postoperatively. Another patient required reoperation 5-months after operation due to additional ABF. No pseudoaneurysms or graft-related complications were observed in the remaining patients. In patients with ABF, performance of operations as soon as possible after onset and minimal dissection of adherent lung tissue appear to improve outcomes. PMID- 15111199 TI - Acute respiratory insufficiency and giant coronary artery aneurysm with fistula. AB - Several causes of acute respiratory insufficiency have been reported in the literature. We describe a case in which it was caused by a rare combination of a giant right coronary artery aneurysm with fistula that occurred in a 56-year-old woman with concomitant congenital hypothyroidism. Diagnostic tools, differential diagnosis, surgical techniques, and follow-up at 1 year are discussed. PMID- 15111200 TI - Resolution of pulmonary arteriovenous fistula by redirection of hepatic venous blood. AB - We report an 8-year-old girl whose unilateral pulmonary arteriovenous fistula (PAVF) disappeared after we redirected hepatic venous blood to the affected lung. This case provides strong evidence that formation of a PAVF depends only on the presence or absence of hepatic venous blood in the pulmonary circulation. PMID- 15111201 TI - Simultaneous repair of pectus excavatum and congenital heart defect in adults by using the convex bar. AB - Simultaneous repair of pectus excavatum and cardiac lesions remains technically difficult. In adults, most repairs of pectus deformity and heart lesions have been performed through long incisions, sternal splits, excision of deformed cartilages, and sternal turnover, which can result in poor cosmetic appearance because of sternal devascularization. We performed concomitant repair of pectus excavatum and an atrial septal defect through a short midline incision in an adult. The sternum was fixed by using absorbable plates and screws and was supported by a convex steel bar. The cosmetic appearance remained excellent after the operation. The technique and a review of the literature are included. PMID- 15111202 TI - Idiopathic tracheal and bronchial stenosis in two pediatric patients. AB - Idiopathic tracheal stenosis, a rare benign chronic inflammatory and fibrotic entity, has previously been reported in the literature in adults, but never in children. Here we report two pediatric cases, one with the rarer condition of idiopathic bronchial stenosis, and we discuss the management and histopathologic findings. PMID- 15111203 TI - A case of primary diffuse tracheobronchial amyloidosis. AB - Tracheobronchial amyloidosis (TBA) is an uncommon localized form of amyloidosis with fewer than 150 reported cases in the literature. We report a case of primary diffuse tracheobronchial amyloidosis who presented with cough, wheezing, recurrent and progressive dyspnea as well as hemoptysis. Though there is no universally accepted treatment for the localized form, this patient was successfully treated with three sessions of bronchoscopic resection and adjuvant steroids. This report will review the various types of amyloidosis of the respiratory tract and their clinical features, in addition to discussing the different available treatment modalities for TBA. PMID- 15111204 TI - Successful thoracoscopic debridement of descending necrotizing mediastinitis. AB - Descending necrotizing mediastinitis results from odontogenic, deep neck infection that spreads along fascial planes into the mediastinum. Although the optimal surgical approach remains controversial, nearly half of the cases require mediastinal debridement by thoracotomy. We report a case of successful thoracoscopic debridement for descending necrotizing mediastinitis due to odontogenic infection that failed to be drained by transcervical approach. Because of less invasiveness as compared with standard thoracotomy, the thoracoscopic approach should be used as early as possible in case of unsuccessful transcervical approach. PMID- 15111205 TI - Surgical resection of recurrent bilateral mediastinal liposarcoma through the clamshell approach. AB - Primary mediastinal liposarcoma is an unusual variant of mediastinal neoplasms. We describe a long-term survivor who underwent repeated operations. After resections through a posterolateral thoracotomy and median sternotomy, a third operation was performed for recurrent bilateral huge tumors through a clamshell incision, and both tumors were removed en bloc. Results of pathologic examination showed that both tumors were well-differentiated liposarcoma. The patient is currently well 16 years after the first operation. Aggressive surgical intervention whenever possible appears to improve the quality of life and prolong the survival of patients with mediastinal liposarcoma. PMID- 15111206 TI - Plasmapheresis as effective treatment for thyrotoxic storm after sleeve pneumonectomy. AB - Thyrotoxicosis is a life-threatening disorder when encountered after a major surgical procedure. Plasma exchange is an occasionally reported alternative treatment to thyroidectomy, iodine ablative therapy, or the administration of thyreostatic drugs. We used plasmapheresis as a lifesaving treatment in a patient with thyrotoxicosis, as encountered after a left-sleeve pneumonectomy, in whom many classic therapies either failed or were not retained. PMID- 15111208 TI - Klippel-Trenauney-Weber syndrome associated with hemoptysis. AB - We report a case of Klippel-Trenaunay-Weber syndrome presenting with hemoptysis. PMID- 15111207 TI - Primary lung tumor mimicking acute type A aortic dissection. AB - A 64-year-old man presented with clinical features and echocardiographic diagnosis of an acute type A dissection. He underwent median sternotomy for definitive surgical treatment. On external examination of the aorta, other intrapericardial structures, and the right lung, it was evident that the patient had an advanced lung tumor. This was confirmed by frozen-section and histopathologic examinations. Epiaortic scanning showed beyond doubt the presence of a mobile intraaortic mass that had misled us in making the preoperative diagnosis of an acute type A dissection. PMID- 15111209 TI - Pulmonary inverted Schneiderian papilloma. AB - Pulmonary inverted Schneiderian papilloma (ISP) is a very rare entity. We herein describe a case that, for its clinical and pathologic features, proved to be challenging. PMID- 15111210 TI - Smoking-related greater vessel stenoses and bilateral axillary artery revascularization. PMID- 15111211 TI - An unusual ectopic calcification in the aorta. PMID- 15111212 TI - Creation of a diaphragm patch to facilitate placement of the AbioCor implantable replacement heart. AB - One of the concerns when placing a total artificial heart is whether the device will fit in the thoracic cavity without impinging on vital structures. We report the creation of a patch in a recipient of the AbioCor Implantable Replacement Heart that allowed for an appropriate fit of the device without adversely affecting pulmonary and hemodynamic functions. PMID- 15111213 TI - Intraabdominal placement of a Novacor ventricular assist system by using an expanded polytetrafluoroethylene pouch. AB - The location of the pump pocket for the left ventricular assist system is of great importance because of its direct relation to subsequent complications. The Novacor device has usually been implanted at the space behind the rectus muscle, which leads to occasional bleeding from raw muscle surfaces and to subsequent pocket infection. Although intraabdominal placement causes less bleeding and is more refractory to later pocket infection, direct contact with the visceral organs sometimes causes serious complications. We constructed a pouch from an expanded polytetrafluoroethylene sheet, and this enabled intraabdominal placement of the Novacor device. PMID- 15111214 TI - Coronary ostial enlargement to prevent stenosis after prosthetic aortic valve replacement. AB - Iatrogenic left and/or right coronary artery ostial stenosis after aortic valve replacement is a rare but life-threatening complication. Although usually related to trauma to the ostium/a during cannulation for administration of cardioplegia, it may be rarely due to direct obstruction by the prosthetic annular ring or stent. We report herein an alternative technique to manage this complication when due to the latter event, successfully utilized at our institution over the last eleven years. PMID- 15111215 TI - Overlapping annuloplasty of the mitral valve in children. AB - Harmonious reduction of the posterior annulus of the mitral valve can be a useful adjunct to obtain complete valve competence in case of annular dilatation. We present a technique with the use of two resorbable sutures that overlap over the middle third of the posterior annulus that was used in 10 children with good short-term results. Resorption of the sutures should permit subsequent normal growth of the mitral valve. If the primary cause of valvular regurgitation was corrected, it can be expected that the repair will remain stable after resorption of the sutures. PMID- 15111216 TI - Thymic tumors. AB - Thymic tumors include thymic carcinoma, which exhibit aggressive behavior, and thymomas, which manifest a more indolent course. Complete resection is the mainstay of treatment, and there appears to be little benefit to partial resection. Postoperative radiotherapy may be useful in incompletely resected patients. Preoperative chemotherapy appears to increase the rate of complete resection and survival of patients with a stage III or IVa thymoma and should strongly be considered in such cases. PMID- 15111217 TI - The first successful repair of postinfarction ventricular septal defect: influence of good fortune, team work, and an inquisitive mind. AB - Research in our institution identified the first case in which cardiopulmonary bypass was utilized. By happenstance, it turned out to be the presumed first repair of postinfarction ventricular septal defect. The circumstances of this interesting case are presented and discussed in the context of our surgical heritage. PMID- 15111218 TI - Mentoring clinical trials in thoracic and cardiovascular surgery: a new role for the Society of Thoracic Surgeons. PMID- 15111219 TI - Increased border-zone stress in bulging ventricular aneurysm. PMID- 15111221 TI - Is routine preoperative mediastinoscopy indicated in clinical stage I non-small cell lung cancer? PMID- 15111222 TI - Video-assisted extended thymectomy in patients with thymoma by lifting the sternum: is it safe? PMID- 15111224 TI - Do we need more trials of postoperative radiotherapy after esophagectomy? PMID- 15111225 TI - Efficacy of the "box" lesion pattern in the treatment of atrial fibrillation in the presence of mitral valve disease. PMID- 15111227 TI - Effects of time-order, interstimulus interval, and feedback in duration discrimination of noise bursts in the 50- and 1000-ms ranges. AB - Participants compared durations of paired white-noise bursts, with interstimulus interval (ISI) 100, 300, 900, or 2700 ms, presented in the order standard (St) comparison (Co) or Co-St. St was 50 or 1000 ms, and 75% difference thresholds for "longer" and "shorter" judgments were estimated. In Experiment 1 feedback was given; in Experiment 2, with ISIs 900 and 2700 ms, there was no feedback. For St=1000 ms, the just noticeable difference (JND) in noise duration was generally smaller with the order St-Co than with Co-St; for St=50 ms, the JND relation was the opposite. JNDs increased with shorter ISIs. Time-order errors were positive for St=50 ms and negative for St=1000 ms, and approached zero for longer ISIs. Using Hellstrom's sensation-weighting (SW) model, the ratio of the stimulus weights for the first and second burst was estimated; this ratio was generally >1 for St=50 ms and <1 for St=1000 ms. JNDs were smaller with feedback than without; the greatest reduction was found for St=1000 ms and an ISI of 2700 ms with the order Co-St, possibly because feedback increased participants' attention to the first stimulus. These results demonstrate the sensitivity of discrimination measures for long as well as brief durations to the factors of ISI, presentation order, and feedback. They also suggest different modes of stimulus processing for short and long durations. PMID- 15111228 TI - The interaction between stop signal inhibition and distractor interference in the flanker and Stroop task. AB - In the present study, two experiments were conducted to investigate the interaction between the behavioral inhibition, measured by the stop signal task, and distractor interference, measured by the flanker task and the Stroop task. In the first experiment, the stop signal task was combined with a flanker task. Analysis revealed that participants responded faster when the distractors were congruent to the target. Also, the data suggest that it is more difficult to suppress a reaction when the distractors were incongruent. Whether the incongruent distractor was part of the response set (i.e. the distractor could also be a target) or not, had no influence on stopping reactions. In the second experiment, the stop signal task was combined with a manual version of the Stroop task and the degree of compatibility was varied. Even though in the second experiment of the present study interference control is differently operationalized, similar results as in the first experiment were found, indicating that inhibition of motor responses is influenced by the presentation of distracting information that is not part of the response set. PMID- 15111229 TI - Automatic non-associative semantic priming: episodic affective priming of naming responses. AB - Affective priming for associatively unrelated prime-target pairs was investigated using (a) the naming task, (b) a short stimulus onset asynchrony (250 ms), and (c) primes that had acquired their affective connotation during a differential evaluative conditioning procedure. Despite the fact that the primes and the targets were related on the affective dimension only, significant priming emerged. This finding indicates that mere affective overlap is sufficient to produce automatic priming. As such, our results are in line with theoretical accounts of automatic priming that are based on semantic relatedness. PMID- 15111230 TI - Intelligence and individual differences in becoming neurally efficient. AB - Physiological approaches to human psychometric intelligence have shown a higher neural efficiency (i.e. less cortical activation) during cognitive performance in brighter subjects. The main aim of this study was to explore the relationship between intelligence and cortical activation patterns in the framework of the learning test concept. In 27 participants we assessed the topography and extent of cortical activation by means of event-related desynchronization (ERD) during reasoning tests in a pre-test--training--post-test design and related it to psychometric intelligence (measured by the German Leistungs-Pruf-System, LPS). Significant associations between intelligence and cortical activation patterns were exclusively found at anterior (frontal) recording sites, which corroborates the central role of the frontal lobe for higher-order cognitive functions. The hypothesized negative intelligence-activation correlation was observed only after the training, i.e. in the post-test, but not in the pre-test. More important, the decrease in cortical investment from pre-test to post-test correlated negatively with intelligence, indicating that the higher the subjects' general mental ability the larger the decrease in the amount of cortical activation. These findings suggest intelligence-related individual differences in becoming neurally efficient. PMID- 15111231 TI - The influence of visual symmetry on the encoding of objects. AB - Two experiments examined how symmetry affects the visual encoding of simple flat objects (lamellae). Experiment 1 showed that subjects encode a lamella's shape as both a facsimile and its enantiomorph. In Experiment 2 the errors made when responding to symmetrical and asymmetrical stimuli showed that there were three factors affecting the reproduction of a symmetrical stimulus. These are, in order of decreasing influence, (i) the orientation of the axis of symmetry, (ii) the presence of overall symmetry (irrespective of orientation), and (iii) identity of the orientation of the elements of the response in relation to those of the stimulus. They also show that symmetry is likely to be reproduced even where the reproduction is not of the same shape as the original stimulus. The relevance of these results to representation of objects is discussed. PMID- 15111232 TI - Play it again: did this melody occur more frequently or was it heard more recently? The role of stimulus familiarity in episodic recognition of music. AB - Episodic recognition of novel and familiar melodies was examined by asking participants to make judgments about the recency and frequency of presentation of melodies over the course of two days of testing. For novel melodies, recency judgments were poor and participants often confused the number of presentations of a melody with its day of presentation; melodies heard frequently were judged as have been heard more recently than they actually were. For familiar melodies, recency judgments were much more accurate and the number of presentations of a melody helped rather than hindered performance. Frequency judgments were generally more accurate than recency judgments and did not demonstrate the same interaction with musical familiarity. Overall, these findings suggest that (1) episodic recognition of novel melodies is based more on a generalized "feeling of familiarity" than on a specific episodic memory, (2) frequency information contributes more strongly to this generalized memory than recency information, and (3) the formation of an episodic memory for a melody depends either on the overall familiarity of the stimulus or the availability of a verbal label. PMID- 15111233 TI - Sertoli cell grafts for Huntington's disease. An opinion. AB - The role of inflammation in CNS diseases is controversial, but growing evidence suggests that anti-inflammatory agents can minimize and/or prevent neural degeneration and its associated behavioral consequences. Sertoli cells can be grafted into the CNS to locally deliver molecules with known trophic and anti inflammatory effects on the surrounding tissue. When Sertoli cells are grafted into the 3-nitropropionic acid (3-NP) model of Huntington's disease the protective effects are quite similar to those obtained using systemic treatments with NSAIDS (Salzberg-Benhouse et al., J. Pharmacol. Exp. Ther. 306:218-228, 2003). While these data alone do not provide unequivocal support for the notion that Sertoli cell grafts exert their beneficial effects via modulating local inflammation, they do provide an interesting convergence between data sets. The benefits of Sertoli cell grafts should be more thoroughly examined in animal models of inflammation. PMID- 15111234 TI - Effects of the DT-diaphorase inhibitor dicumarol on striatal monoamine levels in L-DOPA and L-deprenyl pre-treated rats. AB - It has been proposed that DT-diaphorase plays a strategic role as a neuroprotective enzyme for monoamine neurons, perhaps together with monoamine oxidase (MAO). Thus, we investigated the long-term effects produced by DT diaphorase inhibition with dicumarol injected unilaterally into the medial forebrain bundle (MFB) on monoamine and metabolite levels, alone, or following dopamine loading with 3,4-dihydroxyphenyl-L-alanine (L-DOPA) or MAO inhibition with L-deprenyl. Monoamine levels were assayed in aliquots from tissue samples from right and left striatum, including both dorsal and ventral regions. Dicumarol alone produced increases in 5-hydroxytryptamine (5-HT) and 5 hydroxyindoleacetic acid (5-HIAA), but not in dopamine and metabolite levels when assayed two weeks later. However, following preloading with L-DOPA (3 x 25 mg/kg s.c. 7, 4 and 1 h before surgery), a long-lasting bilateral increase in dopamine and metabolite levels was observed after dicumarol. No effect was observed on dopamine, 5-HT and metabolite levels after L-deprenyl (3 x 10 mg/kg, s.c.) alone, but the levels were unilaterally increased when L-deprenyl was followed by dicumarol. The same result was produced when both L-deprenyl and dicumarol were injected simultaneously into the same brain region. In conclusion, the present study shows that intracerebral inhibition of DT-diaphorase produces long-term changes in 5-HT, but also in dopamine metabolism when DT-diaphorase inhibition is combined with MAO inhibition by systemic or intracerebral treatment with L deprenyl. It is suggested that both MAO and DT-diaphorase have to be inhibited for inducing long-term changes in monoamine metabolism. Thus, DT-diaphorase is an enzyme to be taken into account when L-DOPA is used to treat Parkinson's disease, or when an MAO-inhibitor is used to treat depression. PMID- 15111236 TI - Ethanol effects on dentate granule cell LTP. AB - In previous studies we identified a lateral hypothalamic area (LHA) sensitive to ethanol, < 5.0 mM, when the perifornical region of the area is perfused with different concentrations of ethanol. Some of these perifornical neurons contain angiotensin (Ang) and project directly to the dentate gyrus where angiotensin is released and inhibits LTP in medial perforant path-dentate granule cell synapses. The AT1 subtype receptor is involved because pretreatment with losartan, an AT1 antagonist, prevents Ang II, diazepam, and ethanol impairment of LTP as well as their effects on behavior. There is a possibility that these effects were not specific to the LHA; but might be attributable to direct effects of ethanol on postsynaptic granule cells due to diffusion of the ethanol in the extracellular space or by the circulatory system. The purpose of the present study was to determine a dose effect of ethanol on LTP in these same synapses when the dentate gyrus was perfused with several different concentrations of ethanol under the same conditions in urethane anesthetized rats. Ethanol was administered directly into the dentate gyrus by means of a fine stainless steel cannula attached approximately 1.0 mm from the tip of the glass capillary recording electrode. Results show that the threshold for ethanol in the dentate is higher by a factor of ten, > 30 mM and < 50 mM; and that at higher doses ethanol can have a direct effect on the LHA; and possibly toxic due to increasing ethanol in the blood circulatory system. PMID- 15111235 TI - Terminally differentiated SH-SY5Y cells provide a model system for studying neuroprotective effects of dopamine agonists. AB - We characterized undifferentiated (UN) and three differentiation conditions of the SH-SY5Y neuroblastoma cell line for phenotypic markers of dopaminergic cells, sensitivity to the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridinium ion (MPP+), the requirement to utilize the dopamine (DA) transporter (DAT) for MPP+ toxicity, and the neuroprotective effects of pramipexole. Cells were differentiated with retinoic acid (RA), 12-O-tetradecanoyl-phorbol-13-acetate (TPA), and RA followed by TPA (RA/TPA). RA/TPA treated cells exhibited the highest levels of tyrosine hydroxylase and DAT but lower levels of vesicular monoamine transporter. The kinetics of [3H]DA uptake and [3H]MPP+ uptake to DAT in RA/TPA differentiated cells were similar to that of rat and mouse caudate putamen synaptosomes. RA/TPA differentiated cells evidenced high sensitivity to the neurotoxic effects of MPP+ (0.03 to 3.0 mM), and the neurotoxic effects of MPP+ were blocked with the DAT inhibitor 1-(2-[bis(4-fluorophenyl)methoxy]ethyl) 4-(3-phenylpropyl)piperazine (GBR 12909). DA-induced cell death was not more sensitive in RA vs RA/TPA differentiated cells and was not inhibited by transporter inhibitors. RA/TPA differentiated cells exhibited 3-fold and 6-fold higher levels, respectively, of DA D2 and D3 receptors than UN or RA differentiated cells. Pretreatment with pramipexole was protective against MPP+ in the RA/TPA differentiated cells but not in undifferentiated or RA differentiated cells. The neuroprotective effect of pramipexole was concentration dependent and dopamine D2/D3 receptor dependent. In contrast, protection by pramipexole against DA was not DA receptor dependent. Further characterization of the neuroprotective effects of DA agonists in this model system can provide unique information about DA receptor dependent and independent mechanisms of neuroprotection. PMID- 15111237 TI - Human immunodeficiency virus type 1 protein gp120 causes neuronal cell death in the rat brain by activating caspases. AB - Human immunodeficiency virus type 1 (HIV-1) infection of the central nervous system is associated with microglia activation and neuronal apoptosis, alterations that are also caused by the HIV-1 envelope glycoprotein 120 (gp120) alone. This study was undertaken to examine the onset of gp120 neurotoxicity, the type of cell death and which cells of the adult rat brain are more sensitive to the toxic action of gp120. Gp120 or vehicle were injected chronically (daily for 3 or 7 days) into the lateral ventricle. Magnetic resonance imaging revealed hypertensive areas in the cortical and hippocampal gray matter in gp120-treated rats 7-10 days after the first injection, suggesting vasogenic edema. This phenomenon was accompanied by an enlargement of the lateral and third ventricles. Immunohistochemical analyses were then carried out to examine the toxic effect of gp120 at a cellular level. Several markers of apoptosis, including activated caspase-3 were observed at both 3 and 7 days throughout brains of gp120-treated rats, especially in the cerebral cortex. In this area, most of the apoptotic cells exhibited a pyramidal shape and were Nissl positive, indicative of neurons. Few non-neuronal cells exhibited signs of apoptosis. The results of the present study support the notion that gp120 is neurotoxic in vivo and provide evidence that gp120 activates a caspase-dependent apoptotic pathway. PMID- 15111238 TI - Chronic memantine does not block 3-nitropropionic acid-delayed ischaemic tolerance in rat hippocampal slices ex vivo. AB - The moderate affinity uncompetitive NMDA receptor antagonist memantine, at concentrations found to be neuroprotective in animal models of chronic excitotoxicity, did not reduce ischaemic tolerance induced chemically with 3 nitropropionic acid (3-NP), but actually tended to enhance this effect ex vivo. Injection of 3-NP (20 mg/kg i.p.)--24 h prior to the in vitro experiment- significantly protected against hypoxia/hypoglycaemia-induced suppression of extracellular field excitatory postsynaptic potentials (fEPSPs) in rat hippocampal slices (62.2% vs. control of 16.8%), whereas 3 days pre-treatment with memantine (20 mg/kg/day--Alzet minipumps) tended to enhance recovery further following 3-NP preconditioning (89.7%). This low dose of memantine had no effect on fEPSPs in the absence of preconditioning. As expected, 3 days pre-treatment with a high dose of (+)MK-801 (dizocilpine; 2 mg/kg/day--Alzet minipumps) tended to reduce ischaemic tolerance following 3-NP preconditioning (45.3%). We conclude that although NMDA receptors do seem to be involved in chemically-induced ischaemic tolerance, semi-chronic pre-treatment with therapeutically-relevant doses of memantine does not block ischaemic tolerance. PMID- 15111239 TI - Trimethyltin-induced neurogenesis in the murine hippocampus. AB - Neurogenesis continues to occur in the mature rodent brain with one of the most prominent sources for new neurons being the subgranular layer (SGL) of the dentate gyrus (DG) in the hippocampus. A number of factors can stimulate this process including synaptic activity and injury. To determine if this process would occur upon a direct injury to the dentate region, we exposed young, 21 day old male CD-1 mice to the hippocampal toxicant, trimethyltin (TMT). An acute i.p. injection of TMT (2 mg/kg) produced extensive damage and loss of dentate granule neurons within 72 h. This active period of degeneration was accompanied by an increase in the generation of progenitor cells within the SGL as identified by BrdU uptake and Ki-67 immunostaining. As additional markers for neurogenesis, both nestin and doublecortin showed increased staining patterns within the blades of the dentate. In these young weanling mice, the level of proliferation was sufficient to significantly repopulate the dentate region by 4 weeks post-TMT, suggesting a high level of regenerative potential. Our data indicate a significant level of neurogenesis occurring during the active process of degeneration and in an environment of microglia activation. The TMT-induced injury offers a model system for further examination of the process of neurogenesis, neural adaptation, and the influence of inflammatory factors and glia interactions. PMID- 15111241 TI - Catecholamine neuron groups in rat brain slices differ in their susceptibility to excitatory amino acid induced dendritic degeneration. AB - We investigated whether specific types of catecholamine neurons were differentially vulnerable to damage induced by excitatory amino acids (EAAs) in vitro in a rat brain slice preparation. Brain slices, 300 micro m thick, were cut horizontally, exposed to either N-methyl-D-aspartate (NMDA) or kainic acid (KA) for 2h, fixed and then cut into thin (30 micro m) sections in the same (horizontal) plane as the slice. The sections were immunolabelled for tyrosine hydroxylase to identify different groups of catecholamine neurons (substantia nigra (SN), paranigral (PN), interfascicular (IF) and hypothalamic A11, A13 and A14) which exhibited prominent dendritic projections in the horizontal plane. Loss of dendrites was used as a sensitive index of damage that precedes the loss of the cell body. Catecholamine neurons differed strikingly in their vulnerability of EAA-induced dendrite degeneration. The most vulnerable were those in the dorsal tier of the SN, whereas the most resistant were those in the hypothalamic A11 group. For example, in the dorsal tier of SN, NMDA (50 micro M) reduced the proportion of neurons with dendrites from 64% (+/- 8% SEM) in controls to 13% (+/- 7%) whereas the majority of A11 neurons (69 +/- 10%) retained their dendrites compared to controls (89% +/- 8%). The other groups of catecholamine neurons exhibited intermediate vulnerability. An essentially similar pattern of differential vulnerability was observed with KA. An understanding of the cellular mechanisms that underlie the particular vulnerability of SN neurons in the slice will aid the discovery of pharmacological therapies to prevent or slow the pathological process in neurodegenerative diseases which involve these neurons. PMID- 15111240 TI - Inhibition of DT-diaphorase potentiates the in vivo neurotoxic effect of intranigral injection of salsolinol in rats. AB - The present study shows that intranigral injection of dicoumarol, a DT-diaphorase inhibitor, potentiates the neurotoxic effect of salsolinol (salsolinol 1.25 nmoles plus dicoumarol 2 nmoles; in 2 microl). Rats treated with dicoumarol plus salsolinol presented a characteristic contralateral rotational behaviour when they were stimulated with apomorphine (0.5 mg/kg, s.c.), similar to rats injected unilaterally with 6-hydroxydopamine (6-OHDA). These rats also exhibited impairment of motor and cognitive behaviours. The results support the hypothesis that DT-diaphorase plays a protective role in the nigrostriatal dopaminergic systems. PMID- 15111242 TI - Harmaline induces different motor effects on facial vs. skeletal-motor systems in alert cats. AB - Harmaline's effects on reflex and classically conditioned eyelid responses and on tremor picked up by a coil attached to the back were measured in alert cats. Harmaline at a dose of 10 mg/kg produced skeletal muscle tremogenic effects that lasted 4h. Back movements presented a tremor-like displacement with a frequency peak at 10 Hz, but lid responses oscillated as in controls, at 20 Hz during both reflex and conditioned eyelid movements, with no increase in oscillation amplitude or frequency. The learning curves of harmaline-injected animals remained as in controls, but eyelid conditioned responses showed longer latencies, and smaller amplitude and peak velocity. Reflex and already-learned eyelid responses were not modified by harmaline. These results imply that neuronal control systems for skeletal-motor and facial responses are differentially affected by harmaline. PMID- 15111243 TI - Acute exposure to organochlorine pesticides does not affect striatal dopamine in mice. AB - The purpose of this study was to evaluate the possible association between the risk of developing Parkinson's disease (PD) and exposure to organochlorine pesticides in the mouse model. Animals were treated with a single subcutaneous injection of either dieldrin (40 and 80 mg/kg) or 2,4-dichlorophenoxyacetic acid (100 and 200 mg/kg, 2,4-D) and levels of dopamine (DA) and DA metabolites were measured in the striatum at the 7-day time point. Dieldrin exposure did not affect the striatal concentrations of DA, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA). Administration of 2,4-D did not produce any changes with the exception of a slight (15%), but statistically significant decrease in DOPAC using the higher dose of the pesticide. No neurochemical signs of dopaminergic injury were found following the combined treatment with either dieldrin or 2,4-D plus diethyldithiocarbamate (DDC), a compound known to potentiate the effects of the dopaminergic toxicant 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP). Furthermore, neither dieldrin nor 2,4-D caused additional damage in animals previously lesioned with MPTP. Data failed to support the hypothesis that acute exposure to organochlorine compounds or synergistic interactions involving these pesticides may cause significant damage to dopaminergic terminals and therefore contribute to nigrostriatal degeneration in PD. PMID- 15111245 TI - Behavioral effects induced by acute exposure to benzo(a)pyrene in F-344 rats. AB - Polycyclic aromatic hydrocarbons (PAHs) are highly persistent environmental pollutants which pose potential adverse effects on human health. Benzo(a)-pyrene (B(a)P) is the prototypical representative of these widely dispersed lipophylic contaminants. (B(a)P) exposure in experimental animals results in an array of tissue- and organ-specific responses including carcinogenicity, teratogenicity, reproductive and immunotoxicity. However, no previous studies have examined the potential neurobehavioral toxicity of B(a)P in vivo. The present study was conducted to investigate the behavioral effects induced by single oral doses of (B(a)P) in 8-week-old male and female F-344 rats. Rats were exposed to 0, 12.5, 25, 50, 100 and 200 mg/kg of B(a)P by oral gavage. Motor activity measurements and the functional observational battery (FOB) were used to assess behavioral changes induced by B(a)P at 2, 4, 6, 8, 10, 12, 24, 48, 72 and 96 h post treatment. Statistical analyses revealed significant (p <0.001) dose, sex and time interactions. (B(a)P) doses ranging from 25 to 200 mg/kg produced a significant suppression (up to 60%) in four motor activity parameters: horizontal activity, total distance, stereotype and vertical activity in both sexes within 2 and 4h of dosing. B(a)P treated male and female animals also showed significant (p <0.001) changes in neuromuscular, autonomic, sensorimotor and physiological functions within 2 and 4h post B(a)P administration except in the 12.5 mg/kg treatment group. The 12.5 mg/kg dose did not produce significant (p > 0.05) behavioral toxicity in either males or females. All treated animals (25-200 mg/kg) recovered from the toxic effects of B(a)P by 72 h. Significant (p < 0.05) gender differences were noted in FOB test measures with males displaying greater sensitivity to B(a)P. These data suggest that motor activity and FOB measurements can be used as indices to detect B(a)P neurotoxicity. PMID- 15111244 TI - Adenosine A(2a) receptor antagonists: potential therapeutic and neuroprotective effects in Parkinson's disease. AB - The most effective treatment of Parkinson's disease (PD) is, at present, the dopamine precursor L-3,4-dihydroxyphenylalanine (L-DOPA), however a number of disadvantages such as a loss of drug efficacy and severe side-effects (psychoses, dyskinesias and on-off phenomena) limit long-term effective utilisation of this drug. Recent experimental studies in which selective antagonists of adenosine A(2A) receptors were used, have shown an improvement in motor disabilities in animal models of PD. The A(2A) antagonist [7-(2-phenylethyl)-5-amino-2-(2-furyl) pyrazolo-(4,3-e)-1,2,4-triazolo(1,5-c) pyrimidine] (SCH 58261) potentiated the contralateral turning behavior induced by a threshold dose of L-DOPA or direct dopamine receptor agonists in unilaterally 6-hydroxydopamine (6-OHDA) lesioned rats, an effect accompanied by an increase in Fos-like-immunoreactivity in neurons of the lesioned striatum. Likewise, other A(2A) receptor antagonists such as (3,7-dimethyl-1-propargylxanthine) (DMPX), [E-8-(3,4-dimethoxystyryl)-1,3 dipropyl-7-methylxanthine] (KF 17837) and [E-1,3-diethyl-8(3,4-dimethoxystyryl-7 methyl-3,7-dihydro-1H-purine-2,6-dione] (KW 6002) antagonized catalepsy induced by haloperidol or reserpine in the rat, whereas in non-human primate models of PD, KW 6002 reduced the rigidity and improved the disability score of MPTP treated marmosets and cynomolgus monkeys. Moreover, in contrast to L-DOPA, selective A(2A) receptor antagonists administered chronically did not produce dyskinesias and did not evoke tolerance in 6-OHDA and MPTP models of PD. An additional therapeutic potential of adenosine A(2A) antagonists emerged from studies showing neuroprotective properties of these compounds in animal models of cerebral ischemia and excitotoxicity, as well as in the MPTP model of PD. Adenosine A(2A) receptor antagonists by reversing motor impairments in animal models of PD and by contrasting cell degeneration are some of the most promising compounds for the treatment of PD. PMID- 15111247 TI - Lipid peroxidation and aluminium effects on the cholinergic system in nerve terminals. AB - In the present study, we analyzed how aluminium and oxidative stress induced by ascorbate/Fe(2+) affect the mechanisms related with the cholinergic system in a crude synaptosomal fraction isolated from rat brain. [(3)H]Choline uptake, [(3)H]acetylcholine release, membrane potential and Na(+)/K(+)-ATPase activity were determined in the presence or in the absence of aluminium in control conditions and in the presence of ascorbate (0.8 mM)/Fe(2+) (2.5 micro M). The extent of lipid peroxidation was measured by quantifying thiobarbituric acid reactive substances (TBARS). Under oxidizing conditions aluminium increased the formation of TBARS by about 30%, but was without effect when the synaptosomal preparation was incubated in the absence of oxidants. Additionally, aluminium potentiated the inhibition of the high-affinity [(3)H]choline uptake observed following lipid peroxidation and had the same effect on the Na(+)/K(+)-ATPase activity. [(3)H]Acetylcholine release induced by 4-aminopyridine, and membrane potential were not significantly affected under oxidizing conditions, either in the absence or in the presence of aluminium. We can conclude that aluminium, by potentiating lipid peroxidation, affects the uptake of choline in nerve endings. This effect, occurring during brain oxidative injury, might contribute to the cholinergic dysfunction and neuronal cell degeneration known to occur in Alzheimer's disease. PMID- 15111246 TI - Induction of 78 kD glucose-regulated protein (GRP78) expression and redox regulated transcription factor activity by lead and mercury in C6 rat glioma cells. AB - Lead (Pb) and mercury (Hg) are widespread environmental contaminants that induce prominent neural toxicity. Although the brain is not the major Pb and Hg depot in the body, these metals preferentially accumulate in astroglia to exert toxic effects. In this study, we examined the effects of Pb acetate and HgCl(2) on the expression of GRP78, a molecular chaperone in the endoplasmic reticulum (ER) that may provide cytoprotection in response to cellular stresses in the C6 rat glioma cell line. We also evaluated the DNA binding activities of several redox regulated transcription factors in metal-treated cells. Our results showed that mRNA levels of GRP78 were up-regulated by Pb and Hg at 0.1 and 1 micro M, but down-regulated at higher concentrations (10 micro M). GRP78 protein levels increased in a concentration- and time-dependent manner in Pb and/or Hg-treated cells. Pb increased protein binding to the GST- Upsilon a antioxidant/electrophile response element (ARE/EpRE) and to the NF- kappaB consensus binding sequence of the cytomegalovirus 2 (CMB2) promoter, but decreased protein binding to the Ha-ras ARE/EpRE or to the c-fos 12-O tetradecanoyl-phorbol-13-acetate (TPA) response element (TRE). In contrast, Hg activated DNA binding by all redox-regulated transcription factors. These studies shed some light on the molecular mechanisms of Pb and Hg toxicity in C6 rat glioma cells and suggest that GRP78 and oxidative stress may participate in the neurotoxic response to these metals. PMID- 15111249 TI - Administration of FGF-1 through transfected cells alleviates MPTP toxicity in mice. AB - The use of genetically modified cells to deliver growth factors has been proposed as a possible treatment for neurodegeneration, including Parkinson's disease. Here we demonstrate that the implantation of fibroblasts genetically modified to secrete fibroblast growth factor-1 (FGF) increased striatal dopamine concentrations in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mouse model of Parkinson's disease. PMID- 15111248 TI - Effects of sibutramine on the central dopaminergic system in rodents. AB - The effects of sibutramine on central dopaminergic system in rats and mice were examined by neurochemical and behavioral pharmacological methods. Dopamine reuptake inhibition by sibutramine in brain synaptosomes was only 4-5 times stronger than those of amitriptyline and dosulepin, which do not exhibit dopamine uptake inhibition in vivo. Single treatment with sibutramine did not alter the brain content of dopamine and DOPAC. However, similar to methamphetamine and pargyline, sibutramine antagonized methyl-4-pheny-1,2,3,6-tetrahydro-pyridine (MPTP) induced dopamine depletion in mouse brain. In forced swimming tests of reserpinized mice, sibutramine shortened the immobilized time, similar to dopaminergic drugs including nomifensine, bupropion (dopamine-reuptake inhibitor), methamphetamine, SKF 38393 (dopamine D1 agonist), quinpirole (dopamine D2 agonist) and apomorphine (dopamine D1/D2 agonist). In addition, sibutramine caused rotational behavior toward the lesioned side in rats with unilateral lesions of the substantia nigra induced by 6-hydroxydopamine. These results suggest that sibutramine exhibits neurochemical and behavioral dopaminomimetic activity in vivo, which is mediated by dopamine reuptake inhibition by the active metabolites of sibutramine. PMID- 15111250 TI - MPTP-induced apoptosis in the retina of goldfish. AB - Neuronal degeneration observed in the goldfish retina after MPTP administration, displays features of apoptosis, a physiological mechanism of cell death that occurs during development. The ultrastructural features of degenerating retinal neurons, that are seen in the inner nuclear layer two days after intravitreal MPTP administration, are consistent with classic changes observed in the programmed cell death. The DNA strand breaks that characterize apoptotic death are in situ detected by terminal deoxynucleotidyl transferase-mediated dUTP biotin nick-end labeling. PMID- 15111252 TI - Methamphetamine causes depletion of glutathione and an increase in oxidized glutathione in the rat striatum and prefrontal cortex. AB - The administration of methamphetamine to experimental animals results in damage to dopaminergic neurons. The hypothesis that methamphetamine-induced neurotoxicity is mediated by reactive oxygen species was evaluated. It was found that acute administration of methamphetamine (5 and 15 mg kg(-1)) resulted in production of oxidative stress as demonstrated by decreased glutathione and increased oxidized glutathione levels in the rat striatum and prefrontal cortex. These changes in glutathione and oxidized glutathione levels were dose-dependent in striatum but not in prefrontal cortex. In conclusion, the results of present study provide further evidence in support of the notion that oxidative stress may play an important role in the methamphetamine-induced neurotoxicity. PMID- 15111251 TI - Characterization of intracellular aggregates using fluorescently-tagged polyglutamine-expanded androgen receptor. AB - Spinal bulbar muscular atrophy (SBMA) is a classic CAG-repeat neurodegenerative disease. It is caused by expansion of a polyglutamine (polyGln) tract in the androgen receptor (AR). Recent evidence has indicated a potential role for nuclear and cytoplasmic inclusions in the pathogenesis of these diseases. We have used blue and green fluorescently-tagged AR to show that both wild-type (WT) and poly-Gln-expanded full-length AR can form aggregates and that aggregation is not related to cytotoxicity. Twenty to thirty-five percent of all cell types transfected into COS cells showed aggregation containing both amino- and carboxy terminal fluorescent tags. The aggregates reacted with (F39.4.1), an anti-AR antibody and with IC2, an expanded polyGln tract antibody. Western analysis of protein extracts revealed little evidence of proteolysis although some cleavage of the fusion proteins was seen. The general caspase inhibitor, Z-DEVD-FMK, did not affect aggregation in either wild type or polyGln-expanded GFP-AR transfected cells. Surprisingly, addition of Mibolerone a synthetic androgen significantly decreased inclusion formation in both WT and polyGln-expanded AR-transfected cells. Overall, we show that both WT and polyGln expanded full-length AR are found in aggregates and that proteolysis is not a requirement for aggregation. Our results also suggest that toxicity is not related to intracellular aggregation of polyGln expanded AR. PMID- 15111253 TI - Expressions of caspase-3, Tunel, and Hsp72 immunoreactivities in cultured spinal cord neurons of rat after exposure to glutamate, nitric oxide, or peroxynitrite. AB - Although excitotoxic and oxidative stress play important roles in spinal neuron death, the exact mechanisms are not fully understood. We examined cell damage of primary culture of 11 day-old rat spinal cord by addition of glutamate, nitric oxice (NO) or peroxynitrite (PN) with detection of caspase-3, terminal deoxynucleotidyl transferase-mediated dUTP-biotin in situ nick end labeling (TUNEL) or 72 kDa heat shock protein (HSP72). With addition of glutamate, NOC18 (a slow NO releaser) or PN, immunoreactivity for caspase-3 became stronger in the cytoplasm of large motor neurons in the ventral horn at 6 to 24 hr. TUNEL positive nuclei were found in spinal large motor neurons from 24 h and the positive cell proportion greatly increased at 48 h in contrast to the vehicle. On the other hand, the immunoreactivity of HSP72 in the ventral horn was already positive at 0 h, and gradually decreased in the course of time with glutamate, NOC18 or PN than vehicle treatment. In the dorsal horn, the proportion of caspase 3 positive small neurons greatly increased at 6 to 48 h after addition of glutamate. The present results suggest that both excitotoxic and oxidative stress play important roles in the apoptotic pathway in cultured spinal neurons. PMID- 15111254 TI - Tamoxifen eliminates estrogen's neuroprotective effect upon MPTP-induced neurotoxicity of the nigrostriatal dopaminergic system. AB - The capacity for 17-alpha and 17-Beta estradiol to modulate MPTP-induced neurotoxicity of the nigrostriatal dopaminergic system and potential antagonism of this modulation by the anti-estrogen, tamoxifen, were evaluated. Treatment of retired breeder ovariectomized C57/Bl mice with 17-Beta estradiol diminished the amount of striatal dopamine reduction resulting from MPTP treatment with striatal dopamine concentrations of these 17-Beta estradiol treated mice failing to differ significantly from vehicle treated controls. A combined administration of 17-Beta estradiol with tamoxifen abolished this neuroprotective effect of estrogen as striatal dopamine concentrations of this group were significantly lower than vehicle treated controls. Results to 17-alpha estradiol were less effective since striatal dopamine concentrations of these mice following MPTP treatment were significantly decreased as compared with vehicle controls. In contrast to the nigrostriatal dopaminergic system, no statistically significant effects of these treatments were observed upon olfactory bulb dopamine concentrations. Taken together, these results show that 17-Beta, but not an equivalent concentration of 17-alpha estradiol was effective in decreasing striatal dopamine neurotoxicity to MPTP. This effect of 17-Beta estradiol was abolished by tamoxifen. These data have important implications regarding the mechanisms of estrogen-tamoxifen interactions within the nigrostriatal dopaminergic system as well as for clinical applications of tamoxifen within pre-menopausal women. PMID- 15111255 TI - Assessment of neurotoxicity following repeated cremophor/ethanol injections in rats. AB - As the mixture cremophor/ethanol is known to have side-effects affecting the peripheral nervous system, we have assessed its behavioural and morphological neurotoxicity after repeated intraperitoneal injections in male Sprague Dawley rats. Clinical status of the animals was good throughout the experiment and no motor deficits were observed. Nevertheless, sensory testing demonstrated an hyperalgesia and an allodynia to mechanical stimuli, associated to peripheral axon degeneration. PMID- 15111257 TI - Neurotoxicity and substance abuse: further fuel for regulatory dilemma. PMID- 15111256 TI - Neurodegeneration and glia response in rat hippocampus following nitro-L-arginine methyl ester (L-NAME). AB - Hippocampal neurodegeneration and glia response was examined following administration of the nitric oxide synthase inhibitor, Nomega-nitro-L-arginine methyl ester (L-NAME). Male Long-Evans rats received L-NAME (50 mg/kg, ip) either once or twice a day for 4 days. Both dosing schedules decreased NOS-activity by approximately 90%. At 10 and 30 days following cessation of L-NAME (2x/day), moderate neuronal death was evident in CA1-2 pyramidal cells and dentate granule cells. Neurodegeneration was accompanied by increased astrocyte glial fibrillary acidic protein (GFAP) immunoreactivity yet, minimal astrocyte hypertrophy. Microglia response was limited to an increase in ramified microglia at 10 days, returning to normal by 30 days. As early as 4 days post-dosing (2x/day), GFAP mRNA levels were significantly elevated as were mRNA levels for tumor necrosis factor-alpha (TNFalpha), interleukin-1alpha (IL-1alpha), and interleukin 6 (IL 6). No alterations were seen with L-NAME dosing limited to once a day. The co administration of a hippocampal neurotoxicant, trimethyltin (TMT), with the last dose of L-NAME (2x/day), produced an additive response pattern of neuronal degeneration including both CA1-2 and CA3-4 pyramidal neurons accompanied by TMT induced astrocyte hypertrophy and prominent microglia reactivity. This was preceded by elevations in mRNA levels for GFAP, TNFalpha, IL-1alpha, and IL-6 similar to those seen with each substance alone. These data suggest that high levels of L-NAME can produce a pro-inflammatory environment in the brain and that neurodegeneration and neuroglia responses in the hippocampus can be induced by an alteration in the balance and regulation of local nitric oxide levels. PMID- 15111258 TI - Cognitive effects of neurotoxic lesions of the nucleus basalis magnocellularis in rats: differential roles for corticopetal versus amygdalopetal projections. AB - The cholinergic hypothesis states that cholinergic neurons of the basal forebrain nucleus basalis magnocellularis (nbm) that project to cortical and amygdalar targets play an important role in memory. Biochemical studies have shown that these target areas are differentially sensitive to different excitotoxins (e.g., ibotenate vs. quisqualate). This observation might explain the finding from many behavioural studies of memory that different excitotoxins affect memory differentially even though they produce about the same level of depletion of cholinergic markers in the cortex and similar cortical electrophysiological effects. Thus, the magnitude of mnemonic impairment might be related to the extent of damage to cholinergic projections to the amygdala more than to the extent of damage to corticopetal cholinergic projections. This explanation might similarly apply to the observation that the immunotoxin 192 IgG-saporin produces mild effects on memory when injected into the nbm. This is because it damages cholinergic neurons projecting to the cortex but not those projecting to the amygdala. Studies comparing the effects on memory of ibotenic acid vs. quisqualic acid lesions of the nbm are reviewed as are studies of the mnemonic effects of 192 IgG-saporin. Results support the cholinergic hypothesis and suggest that amygdalopetal cholinergic neurons of the nbm play an important role in the control of memory. PMID- 15111259 TI - Role of the endogenous cannabinoid system as a modulator of dopamine transmission: implications for Parkinson's disease and schizophrenia. AB - The endogenous cannabinoid system is a new signaling system composed by the central (CB1) and the peripheral (CB2) receptors, and several lipid transmitters including anandamide and 2-arachidonylglycerol. This system is the target of natural cannabinoids, the psychoactive constituents of Cannabis sativa preparations (marijuana, hashish). Acute and chronic cannabis exposure has been associated with subjective feelings of pleasure and relaxation, but also to the onset of psychiatric syndromes, a decrease of the efficacy of neuroleptics and alterations in the extrapyramidal system regulation of motor activity. These actions point to a tight association of the cannabinoid system with the brain dopaminergic circuits involved in addiction, the clinical manifestation of positive symptoms of schizophrenia and Parkinson's disease. The present work discusses anatomical, biochemical and pharmacological evidences supporting a role for the endogenous cannabinoid system in the modulation of dopaminergic transmission. Cannabinoid CB1 receptors are present in dopamine projecting brain areas. In primates and certain rat strains it is also located in dopamine cells of the A8, A9 and A10 mesencephalic cell groups, as well as in hypothalamic dopaminergic neurons controlling prolactin secretion. CB1 receptors co-localize with dopamine D1/D2 receptors in dopamine projecting fields. Manipulation of dopaminergic transmission is able to alter the synthesis and release of anandamide as well as the expression of CB1 receptors. Additionally, CB1 receptors can switch its transduction mechanism to oppose to the ongoing dopamine signaling. Acute blockade of CB1 receptor potentiates the facilitatory role of dopamine D2 receptor agonists on movement. CB1 stimulation results in sensitization to the motor effects of indirect dopaminergic agonists. The dynamics of these changes indicate that the cannabinoid system is an activity dependent modulator of dopaminergic transmission, an hypothesis relevant for the design of new therapeutic strategies for dopamine-related diseases such as the psychosis and Parkinson's disease. PMID- 15111260 TI - The developing cholinergic system as target for environmental toxicants, nicotine and polychlorinated biphenyls (PCBs): implications for neurotoxicological processes in mice. AB - During neonatal life, offspring can be affected by toxic agents either by transfer via mother's milk or by direct exposure. In many mammalian species the perinatal period is characterized by a rapid development of the brain - "the brain growth spurt" (BGS). This period in the development of the mammalian brain is associated with numerous biochemical changes that transform the feto-neonatal brain into that of the mature adult. In rodents, the cholinergic transmitter system undergoes a rapid development during the neonatal period, a time when spontaneous motor behaviour also reaches peak activity. We have observed that low dose exposure to environmental toxicants such as nicotine, polychlorinated biphenyls (PCBs) and polybrominated diphenylethers (PBDE, flame retardants) during the "BGS" can lead to irreversible changes in adult brain function in the mouse. The induction of persistent effects on behaviour and cholinergic nicotinic receptors in the adult animal appears to be limited to a short period during neonatal development. Furthermore, the neurotoxic effects were shown to develop over time, indicating a time-response/time-dependent effect. This indicates that environmental toxicants, such as nicotine, PCBs and probably PBDEs, might be involved in the slow, implacable induction of neurodegenerative disorders and/or interfere with normal aging processes. PMID- 15111261 TI - The effect of prenatal cocaine exposure on the stress response of adult mice. AB - The neurotoxic consequences of intrauterine exposure to drugs of abuse, including cocaine, may include compromised fetal brain development with associated lasting behavioral alterations. Some infants exposed to cocaine in utero demonstrate impairments in reactivity and altered behavioral responses to stressful conditions. Alterations in arousal regulation can impact on socialization, adaptation, and educability. Moreover, such alterations may render cocaine exposed children more vulnerable to the adverse developmental impact of stressful situations, with implications for subsequent behavior and psychopathology. Animal models facilitate the independent analysis and identification of genetic, intrauterine, and postnatal environmental factors in contributing to cocaine induced alterations in behavioral and neurochemical responses to stressors. Utilizing a prenatal mouse model of gestational cocaine exposure we have identified a behavioral alteration evident as decreased duration of footshock induced immobility termed "freezing" in cocaine-exposed adults as compared with controls. However, this attenuated behavioral response was not accompanied by demonstrable alterations in corticosterone response, nor was the corticosterone response altered in cocaine-exposed adults following a more protracted restraint induced stress. The dissociation of these behavioral and neurochemical indices of altered response to stressors may provide insights regarding brain mechanisms underlying alterations in behavioral reactivity to stressful conditions following in utero cocaine exposure. In addition, this preclinical study may have implications for improved diagnostics and therapeutics for infants and children exposed to cocaine in the womb. PMID- 15111262 TI - Long term neurodevelopmental and behavioral effects of perinatal life events in rats. AB - Modern neurosciences are now able to open new avenues concerning an experimental approach to clinical neurosciences and psychiatry. Detection and prediction of potential vulnerabilities such as behavioral disturbances and neurodegenerative diseases, are urgent tasks leading to prevention that must be encouraged in parallel to the enormous efforts displayed for treatments. Besides possible genetic origins of diseases, environmental factors are now coming under scrutiny, and especially deleterious and challenging life events and stress occurring during prenatal and postnatal critical periods may orient brain functions towards deleterious developments. The hypothesis that will be examined is that early events might be at the origin of pathological transformations and symptoms after long periods of apparent normal abilities and behavioral homeostasis. We used models of prenatal stress and postnatal manipulations such as cross-fostering. It will be demonstrated that such events induce long-term changes, cognitive and emotional modifications appearing first, when offspring are adults, followed by cognitive defects later in life. Increased sensitivity of the hypothalamic pituitary-adrenal axis (HPA), the endocrine system controlling the secretion of stress hormones (corticoids), appears to be a major element of pathogenesis. HPA axis dysfunction appears very early after birth (3 days) and lasts for months. Cumulative exposure to high levels of hormones seems to be detrimental for some brain regions, especially the hippocampus and major neurotransmitter systems such as dopamine neurons. We evidenced that neuronal modifications in hippocampal region are correlated with behavioral and cognitive defects, relating environment, stress in early life, hormonal changes, long-term neuropathological processes and impaired cognition in aging. Moreover appears in offspring, when adults, a proneness to engage in drug dependence. These data emphasize the need to consider early environmental life events as etiological factors for delayed neuropsychiatric disturbances, neurodegenerative defects included. Moreover, they strengthen the interest for a longitudinal approach to promote experimental psychopathology. PMID- 15111263 TI - Experimental studies on 3,4-methylenedioxymethamphetamine (MDA, "ecstasy") and its potential to damage brain serotonin neurons. AB - A number of drugs that fall into the broad category of "ring-substituted amphetamines" have been found to be neurotoxic toward brain monoamine neurons in animals. Several of these drugs, including 3,4-methylenedioxymethamphetamine (MDMA, "Ecstasy") and methamphetamine ("speed") and fenfluramine ("Pondimin") have been used or abused by humans. A growing body of evidence indicates that humans, like animals, are susceptible to substituted amphetamine-induced neurotoxic injury, and that consequences of this injury can be subtle. This article will review the effects of ring-substituted amphetamine analogs on brain monoamine neurons, using MDMA as the prototype. Studies documenting MDMA neurotoxic potential toward brain serotonin (5-HT) neurons in animals are summarized first. Human MDMA studies are then discussed, beginning with a consideration of methodological challenges in evaluating the status of 5-HT neurons in the living human brain. Recent findings indicating possible functional alterations in brain serotonergic systems in humans with a history of extensive MDMA exposure are then presented, including some new findings on sleep and personality in abstinent MDMA users. PMID- 15111264 TI - Neurotoxicity of methamphetamine and methylenedioxymethamphetamine. PMID- 15111266 TI - Drug addiction: functional neurotoxicity of the brain reward systems. AB - Drug addiction is a chronic relapsing brain disorder characterized by a compulsion to take a drug with loss of control over drug intake. The hypothesis under discussion here is that chronic drug use produces long-lasting dysfunctions in neurons associated with the brain reward circuitry, and this "functional neurotoxicity" of drugs of abuse leads to vulnerability to relapse and continued drug dependence. Several sources of reinforcement are associated with various components of the drug addiction cycle and much progress has been made in identifying the midbrain-basal forebrain neural elements involved in the positive reinforcing effects of drugs of abuse and more recently in the neural elements involved in the negative reinforcement associated with drug addiction. Key elements for the acute reinforcing effects of drugs of abuse include a macrostructure in the basal forebrain called the extended amygdala that contains parts of the nucleus accumbens and amygdala and involves key neurotransmitters such as dopamine, opioid peptides, serotonin, GABA, and glutamate. Withdrawal from drugs of abuse is associated with subjective symptoms of negative affect and dysregulation of brain reward systems involving some of the same neurochemical systems implicated in the acute reinforcing effects of drugs of abuse. In addition, the functional toxicity of acute withdrawal is accompanied by recruitment of the brain stress neurotransmitter system corticotrophin-releasing factor. During more prolonged abstinence, post-acute withdrawal, evidence is accumulating of continued dysregulation of the neural systems associated with drug reinforcement and stress, regulation that may represent more subtle but persistent functional neurotoxic effects of chronic drug use and could be responsible for long-lasting vulnerability to relapse. Such functional neurotoxicity could be hypothesized to lead to a change in set point for drug reward that may represent an allostatic state contributing to vulnerability to relapse and re-entry into the addiction cycle. Elucidation of the specific neuropharmacological changes contributing to this prolonged functional neurotoxicity will be the challenge of future research on the neurobiology of drug addiction. PMID- 15111267 TI - Using claims data to examine patients using practice-based Internet communication: is there a clinical digital divide? AB - BACKGROUND: Practice-based Internet communication allows patients to obtain health information, ask questions, and submit requests through a personalized Web site. While such online tools also bring great promise for educating patients with the goal of fostering behavior change, it is important to examine how individuals currently using such services differ from those who do not. OBJECTIVE: The study used administrative information to characterize a population of patients communicating with a medical practice through the Internet during the end of 1999 and through 2000. METHODS: Patient claims data generated during clinical encounters from January 1999 through May 2000 were examined to measure the relationship between patient demographics, frequency of visits, specific acute diagnoses, and specific chronic diagnoses and the use of online communication with the practice. RESULTS: Ten percent of patients, and 13.2% of patients 18 years or older, used the practice Web site. There were differences in use of the practice Web site by age and insurance status, but not by gender. Use of the practice Web site was similar or higher among patients having a diagnosis for a variety of acute and chronic conditions compared to those not having such a diagnosis. Patients with more clinic visits were more likely to use the Web-based service. CONCLUSIONS: Patients using practice-based Internet communication and having significant health risks can be identified through the use of administrative data, presenting an opportunity to test online educational efforts to improve health. PMID- 15111265 TI - Effects of prenatal exposure to cocaine on the developing brain: anatomical, chemical, physiological and behavioral consequences. AB - Earlier studies of human infants and studies employing animal models had indicated that prenatal exposure to cocaine produced developmental changes in the behavior of the offspring. The present paper reports on the results obtained in a rabbit model of in utero exposure to cocaine using intravenous injections (4 mg/kg, twice daily) that mimic the pharmacokinetics of crack cocaine in humans. At this dose, cocaine had no effect on the body weight gain of dams, time to delivery, litter size and body weight or other physical characteristics of the offspring. In spite of an otherwise normal appearance, cocaine-exposed neonates displayed a permanent impairment in signal transduction via the D1 dopamine receptor in caudate nucleus, frontal cortex and cingulate cortex due to an uncoupling of the receptor from its associated Gs protein. This uncoupling in the caudate nucleus was shown to have behavioral consequences in that young or adult rabbits, exposed to cocaine in utero, failed to demonstrate amphetamine-elicited motor responses normally seen after activation of D1 receptors in the caudate. The cocaine progeny also demonstrated permanent morphological abnormalities in the anterior cingulate cortex due to uncoupling of the D1 receptor and the consequent inability of dopamine to regulate neurite outgrowth during neuronal development. Consistent with the known functions of the anterior cingulate cortex, adult cocaine progeny demonstrated deficits in attentional processes. This was reflected by impairment in discrimination learning during classical conditioning that was due to an inability to ignore salient stimuli even when these were not relevant to the task. The impairment in discrimination learning also occurred in an instrumental avoidance task and could be shown to be due to an impairment of cingulothalamic learning-related neuronal coding. It was proposed that the selective loss of D1-related neurotransmission in the anterior cingulate cortex prevented an appropriate activation of GABA neurons and thus a loss of inhibitory regulation that is necessary for processes involved in associative attention. Taken together, these findings suggest that the uncoupling of the D1 receptor from its G protein may be the fundamental source of the anatomic, cognitive and motor disturbances seen in rabbits exposed to cocaine in utero. Moreover, the long-term cognitive and motor deficits observed in the rabbit model are in agreement with the recent reports indicating that persistent attentional and other behavioral deficits may be evident in cocaine-exposed children as they grow older and are challenged to master more complex cognitive tasks. PMID- 15111268 TI - Online consumer surveys as a methodology for assessing the quality of the United States health care system. AB - BACKGROUND: Interest in monitoring the quality of health care in the United States has increased in recent years. However, the policy objectives associated with collecting this information are constrained by the limited availability of timely and relevant data at a reasonable cost. Online data-collection technologies hold the promise of gathering data directly and inexpensively from large, representative samples of patients and consumers. These new information technologies also permit efficient, real-time assessment in such areas as health status, access to care, and other aspects of the care experience that impact health outcomes. OBJECTIVE: This study investigates the feasibility, validity, and generalizability of consumer online surveys to measure key aspects of health care quality in the United States. METHODS: Surveys about the health and health care experiences of a general adult population and of adults with diabetes were administered online and by telephone. The online survey drew from a sample frame of nearly 1 million consumers and used a single e-mail notification. The random digit-dial methodology included 6 follow-up calls. Results from the online sample were compared to the telephone sample and to national benchmark data. RESULTS: Survey responses about quality of care collected using online and telephone methods were commensurate once they were weighted to represent the demographic distribution of the 2000 United States Census. Expected variations in health and health care quality across demographic and socioeconomic groups were largely observed, as were hypothesized associations among quality indicators and other variables. Fewer individuals were required to be contacted to achieve target sample sizes using online versus telephone methods. Neither method yielded representative cohorts of nonwhite individuals. CONCLUSIONS: Conclusions about the level and variations in health care quality in the United States are similar using data collected in this study compared to data collected using other telephone-based survey methods. As is typical for national telephone surveys conducted by the National Center for Health Statistics, stratified sampling and weighting of survey responses is necessary for results to be generalizable. Online methods are more appropriate for understanding health care quality than for conducting epidemiologic assessments of health in the United States. PMID- 15111270 TI - DietPal: a Web-based dietary menu-generating and management system. AB - BACKGROUND: Attempts in current health care practice to make health care more accessible, effective, and efficient through the use of information technology could include implementation of computer-based dietary menu generation. While several of such systems already exist, their focus is mainly to assist healthy individuals calculate their calorie intake and to help monitor the selection of menus based upon a prespecified calorie value. Although these prove to be helpful in some ways, they are not suitable for monitoring, planning, and managing patients' dietary needs and requirements. This paper presents a Web-based application that simulates the process of menu suggestions according to a standard practice employed by dietitians. OBJECTIVE: To model the workflow of dietitians and to develop, based on this workflow, a Web-based system for dietary menu generation and management. The system is aimed to be used by dietitians or by medical professionals of health centers in rural areas where there are no designated qualified dietitians. METHODS: First, a user-needs study was conducted among dietitians in Malaysia. The first survey of 93 dietitians (with 52 responding) was an assessment of information needed for dietary management and evaluation of compliance towards a dietary regime. The second study consisted of ethnographic observation and semi-structured interviews with 14 dietitians in order to identify the workflow of a menu-suggestion process. We subsequently designed and developed a Web-based dietary menu generation and management system called DietPal. DietPal has the capability of automatically calculating the nutrient and calorie intake of each patient based on the dietary recall as well as generating suitable diet and menu plans according to the calorie and nutrient requirement of the patient, calculated from anthropometric measurements. The system also allows reusing stored or predefined menus for other patients with similar health and nutrient requirements. RESULTS: We modeled the workflow of menu-suggestion activity currently adhered to by dietitians in Malaysia. Based on this workflow, a Web-based system was developed. Initial post evaluation among 10 dietitians indicates that they are comfortable with the organization of the modules and information. CONCLUSIONS: The system has the potential of enhancing the quality of services with the provision of standard and healthy menu plans and at the same time increasing outreach, particularly to rural areas. With its potential capability of optimizing the time spent by dietitians to plan suitable menus, more quality time could be spent delivering nutrition education to the patients. PMID- 15111269 TI - A multimedia interactive education system for prostate cancer patients: development and preliminary evaluation. AB - BACKGROUND: A cancer diagnosis is highly distressing. Yet, to make informed treatment choices patients have to learn complicated disease and treatment information that is often fraught with medical and statistical terminology. Thus, patients need accurate and easy-to-understand information. OBJECTIVE: To introduce the development and preliminary evaluation through focus groups of a novel highly-interactive multimedia-education software program for patients diagnosed with localized prostate cancer. METHODS: The prostate interactive education system uses the metaphor of rooms in a virtual health center (ie, reception area, a library, physician offices, group meeting room) to organize information. Text information contained in the library is tailored to a person's information-seeking preference (ie, high versus low information seeker). We conducted a preliminary evaluation through 5 separate focus groups with prostate cancer survivors (N = 18) and their spouses (N = 15). RESULTS: Focus group results point to the timeliness and high acceptability of the software among the target audience. Results also underscore the importance of a guide or tutor who assists in navigating the program and who responds to queries to facilitate information retrieval. CONCLUSIONS: Focus groups have established the validity of our approach and point to new directions to further enhance the user interface. PMID- 15111271 TI - Sex differences in youth-reported depressive symptomatology and unwanted internet sexual solicitation. AB - BACKGROUND: As the number of youths using the Internet regularly increase, so too does the number of youths potentially vulnerable to negative experiences online. Clinicians, policy makers, and parents need to better understand the Internet and factors related to positive and negative experiences online. OBJECTIVE: Primarily to investigate the association between youth-reported depressive symptomatology and unwanted Internet sexual solicitation and secondarily to identify sex differences in related characteristics of affected youth. METHODS: Data from the Youth Internet Safety Survey were analyzed to investigate the association between reported depressive symptomatology and unwanted Internet sexual solicitation. The Youth Internet Safety Survey was a nationally-representative, cross-sectional telephone survey. Youth participants (N = 1501) were English speakers between the ages of 10 and 17 years who had accessed the Internet at least 6 times in the previous 6 months and had resided in the household for at least 2 weeks in the previous year. Eighty-two percent of contacted households agreed to participate. Each participant was asked to indicate whether any of the 9 symptoms of major depression defined by the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) had been present within the previous month. Logistic regression was used to estimate the odds of reporting an unwanted sexual solicitation online for youths with mild or major depressive symptomatology versus no symptomatology. A parsimonious, multivariate model of significant youth characteristics was identified separately for males and females. RESULTS: Youths who report major depressive-like symptoms were 3.5 times more likely (odds ratio, 3.54; 95% confidence interval, 2.19-5.71) to also report an unwanted sexual solicitation online compared to youths with mild/no symptomatology. After adjusting for significant Internet and psychosocial characteristics, male Internet users who report major depressive-like symptomatology were 2.5 times more likely to also indicate an unwanted Internet solicitation (adjusted odds ratio, 2.72; 95% CI, 1.15-6.40); significant differences were not observed among otherwise-similar females. Further, among youths reporting an Internet solicitation (N = 283), youths with major depressive-like symptomatology were twice as likely to report feeling emotionally distressed by the incident compared to youths with mild/no symptomatology (odds ratio, 2.27; 95% CI, 1.03-5.02). CONCLUSIONS: While the majority of youths report positive experiences online, some youths may be more vulnerable to negative experiences. Cross-sectional results indicate that the report of depressive symptomatology is significantly related to the concurrent report of an unwanted Internet sexual solicitation, especially for young males. Future research should focus on parsing out the temporality of events and identifying additional populations of vulnerable youths online. PMID- 15111272 TI - Can clinical trials requiring frequent participant contact be conducted over the Internet? Results from an online randomized controlled trial evaluating a topical ointment for herpes labialis. AB - BACKGROUND: The Internet has tremendous appeal for conducting randomized clinical trials and may be especially applicable to trials requiring frequent participant contact. Trials of cold sore remedies, for example, often require daily clinic visits during outbreaks, imposing substantial burden on participants. An Internet based randomized clinical trial design may reduce this burden, permitting frequent symptom reports with considerably less effort. OBJECTIVE: To evaluate the feasibility of a Web-based randomized clinical trial requiring frequent participant interaction, using a 6-month, double-blind, randomized, placebo controlled pilot trial of a topical ointment containing dioctyl sodium sulfosuccinate (DSS) (Zilex; Meditech Pharmaceuticals, Inc, Scottsdale, Arizona, USA) intended for treatment of recurrent herpes labialis. A secondary objective was to obtain preliminary data on effectiveness outcomes, to assist in planning a fully-powered trial of DSS. METHODS: Adults with physician-confirmed herpes labialis were recruited to apply to the trial. Eligible applicants were randomized to DSS or placebo, mailed to them upon enrolment with instructions to apply topically every 2 hours for the duration of every cold sore outbreak. Participants were instructed to complete online questionnaires at 2-week intervals and, at the initiation of a cold sore, daily "outbreak questionnaires" until outbreak termination. Feasibility outcome measures included trial participant characteristics, frequency of cold sores, participant retention and adherence (to study medication), and data completeness. Treatment effectiveness outcome measures included outbreak duration, days to crust formation, and pain. RESULTS: Of the 292 individuals applying, 182 screened eligible; 32 participants with confirmed herpes labialis enrolled in the trial. 16 were randomized into the verum group and 16 into the placebo group. 29 (91%) participants completed the trial. During the trial, 34 outbreaks were reported among 23 (72%) participants, resulting in a cold sore incidence rate of 19.8 per 100 person-months of observation. Online data were available for 32 outbreaks; the absence of a resolution date made it impossible to accurately calculate the duration of 12 (38%) outbreaks. Although the DSS treatment group had a shorter mean outbreak duration (6.6 vs 7.7 days, P =.2) and fewer mean days to crust formation (3.5 vs 4.9, P =.1), these differences did not reach statistical significance. The DSS group has statistically significant lower mean pain scores (3.1 vs 7.6, P =.04), but participants in this group also consumed more acetaminophen tablets than the placebo group (1.1 versus 0.5, P=.55). Adherence to medication was similar in both groups: 7 (50%) of the verum group reported using the cream as directed compared to 6 (46.2%) in the placebo group; (P =.8). CONCLUSIONS: We efficiently recruited participants and achieved high overall retention rates. However, participant adherence to the daily outbreak visit schedules was low and only 7 (50%) participants used the cream as directed. These limitations could be addressed in future Internet-based studies by using Personal Digital Assistants (PDAs), using reminder devices, and providing incentives. By enhancing participant adherence, clinical trials requiring frequent participant contact may be feasible over the Internet. PMID- 15111273 TI - Online pediatric information seeking among mothers of young children: results from a qualitative study using focus groups. AB - BACKGROUND: Pre-natal and post-natal periods are times when many women actively seek health information from multiple sources, including the Internet. However, little is known about how pregnant women and mothers of young children seek and process online pediatric health information. OBJECTIVE: To explore why and where mothers of young children look for online health information and how they determine if the information they receive is trustworthy. METHODS: Focus groups were conducted in a Southeastern US city to provide an in-depth exploration of web-related behaviors and beliefs among mothers who work inside and outside of the home. Data from the focus groups were coded using deductive and inductive coding schemes and content was analyzed for the existence of themes. RESULTS: Twenty mothers of young children participated in four focus groups. Most participants sought information on the Internet during pregnancy and nearly all sought online information after their child was born, primarily to diagnose or treat pediatric conditions and to seek advice on parenting and development. Participants mainly used commercial information websites for health information and many expressed disdain for commercial product websites. Many also expressed concerns about the reliability of health information on the web and described strategies for determining how much they trust each website. CONCLUSIONS: Women appear to be high information seekers during pregnancy and the first few years following delivery, and this period represents an important window of time for providing online health information. Participants suggested that online information sources and motives for providing online information should be clear in order to increase perceptions of trust. Participants expressed preference for online clinical health information that is presented by clinical professionals, and online parenting advice that is presented from other parents. PMID- 15111274 TI - The Internet as a vehicle to communicate health information during a public health emergency: a survey analysis involving the anthrax scare of 2001. AB - BACKGROUND: The recent public health risks arising from bioterrorist threats and outbreaks of infectious diseases like SARS (Severe Acute Respiratory Syndrome) highlight the challenges of effectively communicating accurate health information to an alarmed public. OBJECTIVE: To evaluate use of the Internet in accessing information related to the anthrax scare in the United States in late 2001, and to strategize about the most effective use of this technology as a communication vehicle during times of public health crises. METHODS: A paper-based survey to assess how individuals obtained health information relating to bioterrorism and anthrax during late 2001. We surveyed 500 randomly selected patients from two ambulatory primary care clinics affiliated with the Brigham and Women's Hospital in Boston, Massachusetts. RESULTS: The response rate was 42%. While traditional media provided the primary source of information on anthrax and bioterrorism, 21% (95% CI, 15%-27%) of respondents reported searching the Internet for this information during late 2001. Respondents reported trusting information from physicians the most, and information from health websites slightly more than information from any traditional media source. Over half of those searching the Internet reported changing their behavior as a result of information found online. CONCLUSIONS: Many people already look to the Internet for information during a public health crisis, and information found online can positively influence behavioral responses to such crises. However, the potential of the Internet to convey accurate health information and advice has not yet been realized. In order to enhance the effectiveness of public-health communication, physician practices could use this technology to pro-actively e-mail their patients validated information. Still, unless Internet access becomes more broadly available, its benefits will not accrue to disadvantaged populations. PMID- 15111276 TI - Experience and attitudes towards information technology among first-year medical students in Denmark: longitudinal questionnaire survey. AB - BACKGROUND: As more and more information technology (IT) resources become available both for support of campus- based medical education and for Web-based learning, it becomes increasingly interesting to map the information technology resources available to medical students and the attitudes students have towards their use. OBJECTIVE: To determine how extensively and effectively information handling skills are being taught in the medical curriculum, the study investigated Internet and computer availability and usage, and attitudes towards information technology among first-year medical students in Aarhus, Denmark, during a five-year period. METHODS: In the period from 1998 to 2002, students beginning the first semester of medical school were given courses on effective use of IT in their studies. As a part of the tutorials, the students were asked to complete a web-based questionnaire which included questions related to IT readiness and attitudes towards using IT in studies. RESULTS: A total of 1159 students (78%) responded. Overall, 71.7% of the respondents indicating they had access to a computer at home, a number that did not change significantly during the study period. Over time, the power of students' computers and the use of e mail and Internet did increase significantly. By fall 2002, approximately 90% of students used e-mail regularly, 80 % used the Internet regularly, and 60 % had access to the Internet from home. Significantly more males than females had access to a computer at home, and males had a more positive attitude towards the use of computers in their medical studies. A fairly constant number of students (3-7 %) stated that they would prefer not to have to use computers in their studies. CONCLUSIONS: Taken together with our experience from classroom teaching, these results indicate optional teaching of basic information technology still needs to be integrated into medical studies, and that this need does not seem likely to disappear in the near future. PMID- 15111275 TI - Swiss community pharmacies' on the Web and pharmacists' experiences with E commerce: longitudinal study and Internet-based questionnaire survey. AB - BACKGROUND: There are multiple ways in which community pharmacies can present themselves on the Internet, e.g., as a platform for drug information or as an advertising platform for their services. OBJECTIVE: To estimate the number of Swiss community pharmacies on the Internet over the period of 32 months (2000 2003), to describe their current e-commerce services, and to explore the experiences and plans these pharmacies have with regard to their Internet presence. METHODS: A longitudinal study was performed to determine the number of Swiss German pharmacies on the Internet by conducting Internet searches in 2000, 2001, and 2003. In April 2002, a cross-sectional Internet-based survey was administered to explore the pharmacies' experiences and plans regarding their Web sites. RESULTS: As of April 2003, 373 (44%) of 852 community pharmacies from the German speaking part of Switzerland were on the Internet. One hundred eighty four listed an e-mail address and were asked to complete a questionnaire. Of the 107 pharmacies answering the survey questions (58% response rate): 46% had been on the Internet for 1 to 2 years; 33% of the Web sites are part of a pharmacy group's Web portal; 31% of the pharmacies plan to expand their Internet appearance in the future; 74% provide e-commerce services, with 81% of those pharmacies filling five or less orders per month; and 12% plan on expanding their e-commerce services in the future. CONCLUSIONS: The number of community pharmacies offering Internet services steadily increased over 32 months. Given the importance of the Internet as a tool for information, communication, and advertising for pharmacy products and services, it can be expected that the increase will continue. Pharmacy-group portals are important promoters of pharmacies on the Internet. For many community pharmacies, Internet portals that provide an Internet presence for the pharmacies and provide regularly-updated content (e.g., health news, tips, drug information) seem to be the most effective solutions. Even though 40% of the pharmacies already offer e-commerce services, these services are still of minor importance. For many pharmacists, the current legal regulations seem to be unclear. Most pharmacies want to maintain their Internet services. PMID- 15111278 TI - Cerebral specialization and the control of oral and limb movements for individuals with Down's syndrome. AB - In this paper we review existing research on cerebral specialization in Down's syndrome (DS) individuals, and we present a preliminary model. The model proposes that the cerebral organization of DS persons is such that there is a dissociation of brain areas responsible for speech perception and the production of complex movement including speech. Research is presented that confirms some initial predictions of the model. PMID- 15111277 TI - Pharmacist computer skills and needs assessment survey. AB - BACKGROUND: To use technology effectively for the advancement of patient care, pharmacists must possess a variety of computer skills. We recently introduced a novel applied informatics program in this Canadian hospital clinical service unit to enhance the informatics skills of our members. OBJECTIVE: This study was conducted to gain a better understanding of the baseline computer skills and needs of our hospital pharmacists immediately prior to the implementation of an applied informatics program. METHODS: In May 2001, an 84-question written survey was distributed by mail to 106 practicing hospital pharmacists in our multi-site, 1500-bed, acute-adult-tertiary care Canadian teaching hospital in Vancouver, British Columbia. RESULTS: Fifty-eight surveys (55% of total) were returned within the two-week study period. The survey responses reflected the opinions of licensed BSc and PharmD hospital pharmacists with a broad range of pharmacy practice experience. Most respondents had home access to personal computers, and regularly used computers in the work environment for drug distribution, information management, and communication purposes. Few respondents reported experience with handheld computers. Software use experience varied according to application. Although patient-care information software and e-mail were commonly used, experience with spreadsheet, statistical, and presentation software was negligible. The respondents were familiar with Internet search engines, and these were reported to be the most common method of seeking clinical information online. Although many respondents rated themselves as being generally computer literate and not particularly anxious about using computers, the majority believed they required more training to reach their desired level of computer literacy. Lack of familiarity with computer-related terms was prevalent. Self reported basic computer skill was typically at a moderate level, and varied depending on the task. Specifically, respondents rated their ability to manipulate files, use software help features, and install software as low, but rated their ability to access and navigate the Internet as high. Respondents were generally aware of what online resources were available to them and Clinical Pharmacology was the most commonly employed reference. In terms of anticipated needs, most pharmacists believed they needed to upgrade their computer skills. Medical database and Internet searching skills were identified as those in greatest need of improvement. CONCLUSIONS: Most pharmacists believed they needed to upgrade their computer skills. Medical database and Internet searching skills were identified as those in greatest need of improvement for the purposes of improving practice effectiveness. PMID- 15111279 TI - Acquisition of route and survey knowledge in the absence of vision. AB - The ability of sighted, blindfolded individuals to navigate while walking was assessed in two types of tasks, one requiring knowledge of a route that previously had been navigated and another requiring more complex spatial inference or computation. A computerized measurement system monitored spatial position. The route tasks included maintenance of a heading, distance and turn reproduction and estimation, and turn production. The inferential task required completion of a multisegment pathway by returning directly to the origin. pathways were replicated at two different scales. Measures for the route knowledge tasks indicated a substantial ability to navigate in the absence of visual cues. Route reproduction performance was particularly accurate despite intrinsic veering tendencies. A substantial increase in error was observed in the pattern-completion task. Errors in pathway completion increased with pathway complexity and were quite similar in the two scales. Correlational data suggested that performance on different route-knowledge tasks reflected differing underlying representations. The completion task led to a high correlation between absolute turn and distance error but had minimal correlations with the route tasks. The data suggest that a survey representation with some degree of scale independence was constructed for use in the pathway completion task. PMID- 15111280 TI - Time-dependent effects of kinesthetic input. AB - Sensory input can be used by the nervous system to control the spatial parameters of motor responses (e.g., distance, velocity, and direction) by initializing these parameters before movement onset and then by adjusting these parameters during movement. Sensory input can also be used to trigger movements. In the experiments reported in this paper, we compared the effects of kinesthetic input on a triggered motor response when the kinesthetic input was generated at different times relative to the onset of the motor response. Human subjects responded to a visual stimulus by intentionally increasing elbow torque to a target level. Kinesthetic input was generated by unexpectedly rotating each subject's elbow 100 ms before the onset of the intentional torque response (early) or coincident with the onset of the intentional torque response (late). The effect of early kinesthetic input on the intentional torque response markedly differed from the effect of late kinesthetic input. The effect of early kinesthetic input was relatively independent of the direction of elbow rotation, had a different dependence on the amplitude of rotation, and required a shorter duration of rotation compared to the effect of late kinesthetic input. These differences in the effects of early and late kinesthetic input might be related to the initialization, triggering, and adjustment of motor responses. PMID- 15111281 TI - Mechanisms underlying accuracy in fast goal-directed arm movements in man. AB - This study investigated how accuracy is attained in fast goal-directed arm movements. Subjects were instructed to make arm extension movements over three different distances in random order, with and without visual feedback. Target width was varied proportionally with distance. Movement time was kept as short as possible, but there were well-defined limits with respect to accuracy. There appeared to be a large relative variability (variation coefficient [VC]) in the initial acceleration. The VC in the distance the hand moved during the acceleration phase was much smaller. This reduction was accompanied by a strong negative correlation between the initial acceleration and the duration of the acceleration phase. Further, the VC in the total distance moved was less than the VC in the distance moved during acceleration. This result indicates asymmetry between the acceleration and the deceleration phase. This is confirmed by the negative correlation between the distance the hand moved during acceleration and the distance it moved during deceleration. Withdrawal of visual feedback had a significant effect on movement accuracy. No differences were found in the parameters of the acceleration phase in the two feedback conditions, however. our results point to the existence of a powerful variability compensating mechanism within the acceleration phase. This mechanism seems to be independent of visual feedback; this suggests that efferent information (efference copies) and/or proprioceptive information is/are responsible for the timing of agonist and antagonist activation. The asymmetry between the acceleration and deceleration phase contributes to a reduction in the relative variability in the total distance moved. The fact that the withdrawal of visual feedback affected movement variability only during the deceleration phase indicates that visual information is used in the adjustment of antagonist activity. PMID- 15111282 TI - Representational guidance of action production in observational learning: a causal analysis. AB - This experiment tested the hypothesis that the number of model presentations and verbal coding of modeled actions affect reproduction accuracy through their effect on cognitive representation. Subjects viewed a complex action pattern either two or eight times with or without verbal coding to highlight the dynamic structure of the component actions and their temporal sequencing. They then received, in order, a recognition test and a pictorial-arrangement test to assess the accuracy of their cognitive representations of the modeled actions. Subsequently, all subjects were tested for their ability to reproduce the action pattern from memory. Results showed that increased exposure to modeled actions enhanced the accuracy of both the cognitive representation and the behavioral reproduction. Verbal coding also increased cognitive and reproduction accuracy, but only when combined with multiple opportunities to observe the modeled actions. A causal analysis confirmed that the effects of multiple exposures and verbal coding were entirely mediated by changes produced in the accuracy of cognitive representation. PMID- 15111283 TI - Intentional switching between patterns of bimanual coordination depends on the intrinsic dynamics of the patterns. AB - Two predictions arising from previous theoretical and empirical work which demonstrated that spontaneous changes of bimanual coordination patterns result from a loss of pattern stability (i.e., a nonequilibrium phase transition) were tested: (a) that the time it takes to intentionally switch from one pattern to another depends on the differential stability of the patterns themselves; and (b) that an intention, defined as an intended behavioral pattern, can change the dynamical characteristics, e.g., the overall stability of the behavioral patterns. Subjects moved both index fingers rhythmically at one of six movement frequencies while performing either an in-phase or antiphase pattern of finger coordination. On cue from an auditory signal, subjects switched from the ongoing pattern to the other pattern. The relative phase of movement between the two fingers was used to characterize the ongoing coordinative pattern. The time taken to switch between patterns, or switching time, and relative phase fluctuations were used to evaluate the modified pattern dynamics resulting from a subject's intention to change patterns. Switching from the in-phase to the antiphase pattern was significantly slower than switching in the opposite direction for all subjects. Both the mean and distribution of switching times in each direction were found to be in agreement with model predictions. movement frequency had little effect on switching time, a finding that is also consistent with the model. Relative phase fluctuations were significantly larger when moving in the antiphase pattern at the highest movement frequencies studied. The results show that, although intentional influences act to modify a coordinative pattern's intrinsic dynamics, the influence of these dynamics on the resulting behavior is always present and is particularly strong at high movement frequencies. PMID- 15111284 TI - Is it possible to prepare the second component of a movement before the first one? AB - Experiment 1 utilized a choice reaction time paradigm to examine whether advance information about the second component of a movement has similar effects upon movement initiation and execution as advance information about the first component. Four stimuli were assigned to four goal keys. Subjects responded with the index finger of their preferred hand. They had to press on of two intermediate keys before pressing the assigned stimulus goal key. Advance information signaled one pair of goal keys in such a way that either the first or the second movement component was unequivocally specified before the response signal appeared. Shorter reaction times resulted when the first component was precued. Further control conditions showed that advance information about the second movement component could not be utilized for movement preparation. Experiment 2 ruled out a perceptual interpretation of this effect. Experiment 3 showed that preparation time for two-component movements are longer than for one component movements. The study permits the conclusions that speeded two-component movements are controlled by motor programs and that advance information about the first movement component is required before the second movement component can be programmed. PMID- 15111285 TI - The role of attention in one-handed catching. AB - The present study investigated the contribution of attention to one-handed catching success. A group of skilled (n = 8) and less skilled (n = 9) male subjects were compared in their ability to process secondary task information while executing a primary one-handed catching task. On 40% of the trials, a secondary visual stimulus (SVS) was presented in the peripheral visual field at predetermined times during the flight of the ball. On these trials, the subject was required to complete the one-handed catch and immediately throw the ball at a stationary target. Less skilled subjects made significantly more catching errors under both normal viewing and dual-task processing conditions. The differences were due to errors of positioning rather than grasping. Positioning of the hand appears to require visual attention regardless of skill level, as both skill groups experienced increased difficulty processing secondary task information as the ball approached the catching hand. PMID- 15111287 TI - Spatial, temporal, and electromyographical variability in human motor control. A symposium held at the University of Michigan, Ann Arbor, February 17-18, 1989. PMID- 15111286 TI - A noted on the relationship between task requirements and the contextual interference effect. AB - Typically, tasks used in past contextual interference experiments had movement, spatial pattern, or timing requirements. The possibility exists that the blocked/random manipulation of only one of these task characteristics contributes to the contextual interference effect. The purpose of the experiment reported here was to test the impact of separate movement and timing tasks on the superior learning of random trained groups. The task for all subjects in the movement condition was to release a start button and knock over a wooden barrier. There were three movement goals to be learned. Half of the subjects in this condition practiced the three movements in a blocked schedule and half practiced them in a random schedule. The subjects in the no-movement condition estimated the same three times by holding down the start button for the appropriate duration. Similarly, these subjects were divided into random and blocked practice groups. All subjects then performed a retention test. Results showed that for the movement condition, the blocked group performed with less error than the random group during acquisition. In retention, however, the random group performed with less error than the blocked group. conversely, for the no-movement condition, there were no differences between the two practice schedule groups during acquisition or during retention for any of the dependent measures. These results indicated that experimental tasks must have some type of movement requirement in order to facilitate learning through the use of random practice schedules. PMID- 15111288 TI - Preload and isometric force variability. AB - Two experiments are reported, which examined the relative contributions of preload (resting force level), change of force, and the time taken to achieve the force in determining isometric force variability. The findings showed that change of force is the strongest determiner of peak force variability but that preload and time to peak force have smaller though systematic effects. A formula that predicts peak force variability is proposed, with preload as an additive effect to the ratio between the change of force level and the square root of time to peak force. These findings confirm that these three impulse variables are significant in predicting force variability and that the impact of rate of force on peak force variability is nonlinear. PMID- 15111289 TI - On the role of knowledge of results in motor learning: exploring the guidance hypothesis. AB - Recent work on the role of knowledge of results (KR) in motor learning has challenged some traditional assumptions. In particular, the guidance hypothesis suggests that there is a detriment to learning when KR guides the learner toward correct performance. Three experiments that explored this hypothesis are reported here. These experiments contrasted the effects of relative frequency of guidance versus the relative frequency of KR. According to the guidance hypothesis, it was predicted that parallel effects of the relative frequency manipulation on motor learning and performance would result under guided and KR conditions. The movement task was a reciprocal timing task that was either paced by a metronome (Experiment 1) or augmented by auditory KR (Experiments 2 and 3). The results of Experiments 1 and 3 revealed a number of parallel effects, thus providing support for the guidance hypothesis. The contrast of Experiments 1 and 2 resulted in a number of dissociable effects, however. These findings are discussed in relation to the potential guiding properties of KR and their impact on motor performance and learning. PMID- 15111290 TI - Effects on prism adaptation of duration and timing of visual feedback during pointing. AB - In two experiments, we investigated the effects of duration of visual feedback of the pointing limb and the time (early to late) in the movement when the limb first becomes visible (timing of visual feedback). Timing, rather than duration of visual feedback, proved to have the greater effect on the relative magnitude of visual and proprioceptive adaptation. Visual adaptation increased smoothly with feedback delay, but corresponding decreases in proprioceptive adaptation underwent an additional sharp change when feedback was delayed until about three fourths of the way to the terminal limb position. These results are consistent with the idea that visual and proprioceptive adaptation are mediated by exclusive processes. Change in the limb position sense (i.e., proprioceptive adaptation) may be produced by visual guidance of the pointing limb, and view of the limb early in the pointing movement seems to be critical for such visual guidance. The limb may be ballistically released as it nears the terminal position, and, thereafter, any opportunity for visual guidance (i.e., view of the limb) is not effective. On the other hand, change in the eye position sense (i.e., visual adaptation) may be mediated by proprioceptive guidance of the eye; the eyes may track the imaged position of the nonvisible limb. Such proprioceptive guidance seems to be solely a function of the distance moved before the limb becomes visible. PMID- 15111291 TI - Tracking with and without target in 6- to 15-year-old boys. AB - It is well documented that, in adults, manual tracking of a visual periodic target is very accurate in a wide variety of experimental conditions. With children between 5 and 9 years of age, however, the response lags significantly behind the stimulus. The first study presented here attempts to described the acquisition of this skill by children between 9 and 15 years of age. Within this age span, adult proficiency in pursuing a rapid target was approached through an improvement in response synchronization. Yet adults were distinguishable from the oldest children by a fundamentally different mode of movement execution: The former maintained a smooth modulation of the proper motor pattern, whereas the latter relied mainly on corrections through visual feedback. The second experiment showed that these different perceptuomotor strategies may be related to the availability of a more accurate and stable motor pattern with age. Young children had difficulties reproducing the stimulus after it was withdrawn, with performance deteriorating as the trial progressed. The initial mismatch and the following drift tended to decrease with age, even though the oldest children's stationary performance was still not as consistent with the target motion as the adult response. PMID- 15111292 TI - Anticipatory postural control in children. AB - Postural responses, triggered by sensory feedback, are present very early in a child's development. The purpose of the present study was to investigate the ability of children to anticipate postural disturbances caused by self-initiated movements and their ability to coordinate anticipatory postural adjustments with movement execution. Children (N = 32) aged 4 to 14 years were asked to stand quietly on a stable force plate and to raise their right arm forward (or backward) to the horizontal position after a visual stimulus. Changes in the center of pressure beneath the feet were recorded before and during the arm raise. The anticipatory (feedforward) postural patterns seen before the arm movement, and noted in a previous study of adults, were present in the youngest of the children (4 years, 2 months). Longer reaction times and inconsistent postural responses (in the anteroposterior direction) suggest that children are less capable than adults of coordinating the anticipated postural adjustment with the forthcoming limb movement, however. In the lateral plane, anticipatory postural responses were initiated more consistently. PMID- 15111293 TI - Practice and assimilation effects in a multilimb aiming task. AB - When two limbs are required to move different distances simultaneously, assimilation effects are shown: The shorter distance limb tends to overshoot the target, whereas the longer distance limb undershoots. The effect of practice on assimilation effects was studied in two experiments, using a simultaneous four limb aiming task. When subjects were required to move their left limbs a shorter distance than the right (5 cm vs. 9 cm), the right limbs moved a lesser distance and had greater variable and overall errors relative to a group required to move all limbs the same distance (9 cm). Practice reduced assimilation effects in the lower limbs, but spatial assimilations were present throughout 125 acquisition trials with KR and 50 no-KR transfer trials, spanning 24 hours. When the upper limbs were required to move a greater distance than the lower limbs (15 cm vs. 9 cm), the lower limbs showed longer distances and increased overall errors early in practice compared to the lower limbs of a group required to move all limbs 9 cm. With practice, the between-group differences decreased, with no assimilation effects shown on the transfer trials. The results suggest that neural crosstalk is greater between left and right sides than between upper and lower limbs. Results are discussed in light of the functional cerebral space model of simultaneous actions. PMID- 15111294 TI - The effect of motor preparation on changes in h reflex amplitude during the response latency of a warned reaction time task. AB - Monosynaptic Hoffman reflexes (H reflexes) were recorded from the soleus muscle during the response latency of a warned reaction time (RT) task that required plantarflexion of the foot. The task was done under four conditions of predictability of the response signal (RS), created by the factorial combination of foreperiod duration (1 and 4 s) and variability (fixed and variable). RT varied systematically with RS predictability and was facilitated in conditions that favored prediction of the RS. The response latency was divided into two successive phases by the onset of reflex augmentation: a premotor phase of constant reflex amplitude and a succeeding motor phase marked by progressively increasing reflex amplitude. Reflex augmentation during the motor phase was coupled more closely to the imminent movement than to the preceding signal to respond. The duration of the premotor phase was unaffected by RS predictability, but the duration of the motor phase (like RT) was shorter when the RS was more predictable. The maximum H reflex amplitude reached during the motor phase was greater when the RS was more predictable. The tonic level of H reflex amplitude during the premotor phase was greater in conditions that made prediction of the RS difficult. A second experiment showed that this difference was present throughout the foreperiod. These results suggest that conditions that favor prediction of the RS enhance motor preparation. changes in motor preparation (which affect RT) affect the processes underlying reflex amplitudes in the premotor phase and throughout the preceding foreperiod, in conditions that make prediction of the RS difficult, appear to reflect heightened general arousal. PMID- 15111295 TI - Assessing and reporting the accuracy of position measurements made with optical tracking systems. AB - The use of optical tracking systems to record human movement is now widespread. Although such systems are convenient and potentially very accurate, they must be used carefully to ensure good data. This paper describes the procedures of calibration and reconstruction of position data in cartesian coordinates and suggests steps to maximize their accuracy. Procedures are proposed for characterizing the accuracy of measurement throughout the experimental workspace, and open discussion of the issue by the research community is invited. PMID- 15111297 TI - Inflammation and remyelination in the central nervous system: a tale of two systems. PMID- 15111296 TI - Ovarian tumorigenesis: a proposed model based on morphological and molecular genetic analysis. AB - The pathogenesis of ovarian carcinoma, the most lethal gynecological malignancy, is unknown because of the lack of a tumor progression model. Based on a review of recent clinicopathological and molecular studies, we propose a model for their development. In this model, surface epithelial tumors are divided into two broad categories designated type I and type II tumors that correspond to two main pathways of tumorigenesis. Type I tumors tend to be low-grade neoplasms that arise in a stepwise manner from borderline tumors whereas type II tumors are high grade neoplasms for which morphologically recognizable precursor lesions have not been identified, so-called de novo development. As serous tumors are the most common surface epithelial tumors, low-grade serous carcinoma is the prototypic type I tumor and high-grade serous carcinoma is the prototypic type II tumor. In addition to low-grade serous carcinomas, type I tumors are composed of mucinous carcinomas, endometrioid carcinomas, malignant Brenner tumors, and clear cell carcinomas. Type I tumors are associated with distinct molecular changes that are rarely found in type II tumors, such as BRAF and KRAS mutations for serous tumors, KRAS mutations for mucinous tumors, and beta-catenin and PTEN mutations and microsatellite instability for endometrioid tumors. Type II tumors include high-grade serous carcinoma, malignant mixed mesodermal tumors (carcinosarcoma), and undifferentiated carcinoma. There are very limited data on the molecular alterations associated with type II tumors except frequent p53 mutations in high grade serous carcinomas and malignant mixed mesodermal tumors (carcinosarcomas). This model of carcinogenesis reconciles the relationship of borderline tumors to invasive carcinoma and provides a morphological and molecular framework for studies aimed at elucidating the pathogenesis of ovarian cancer. PMID- 15111298 TI - Telomere lengths of translocation-associated and nontranslocation-associated sarcomas differ dramatically. AB - Sarcomas can be divided into those with specific translocations displaying monotonous cytomorphology, and those with complex karyotypes and marked cellular pleomorphism. Telomeres contain terminal DNA sequence repeats that maintain chromosomal stability. Telomeres shorten with cell division and may become dysfunctional leading to chromosomal instability. Using a fluorescence in situ hybridization/immunofluorescence method to assess telomere lengths in archival tissues we analyzed these two types of sarcomas using paraffin-embedded primary tumor specimens. Tissues from nine sarcomas with characteristic translocations (two synovial sarcomas, two alveolar rhabdomyosarcomas, two desmoplastic round cell tumors, and one each of infantile fibrosarcoma, myxoid liposarcoma, cellular congenital mesoblastic nephroma) and nine without (four malignant fibrous histiocytomas, two leiomyosarcomas, one pleomorphic rhabdomyosarcoma, one dedifferentiated chondrosarcoma, and one malignant peripheral nerve sheath tumor) were analyzed. In all (nine of nine) cases with specific translocations, which generally have few karyotypic abnormalities, telomere lengths were similar to or reduced compared to surrounding nonneoplastic tissues. In contrast, telomeres in cases lacking specific translocations, which generally contain complex karyotypes, were often found to be dramatically lengthened and heterogeneous. In addition to markedly elongated telomeres, seven of nine (78%) complex cases exhibited large brightly stained regions corresponding to a specific type of promyelocytic leukemia nuclear body found in immortalized cells that maintain telomeres in a telomerase-independent manner [alternative lengthening of telomeres (ALT) pathway]. This phenotype is unlike that of epithelial neoplasms that typically display complex karyotypes with abnormally short telomeres maintained by the enzyme telomerase. The discovery of heterogeneous telomere lengths and evidence of the ALT pathway in the majority of sarcomas with complex karyotypes supports the existence of a telomere maintenance pathway incapable of full karyotypic stabilization in pleomorphic sarcomas. These findings provide additional molecular-genetic evidence supporting the dichotomous grouping of sarcomas into those with characteristic signature translocations without extensive additional karyotypic abnormalities, and those without such signature translocations that typically display very complex karyotypes, and point to telomere dysfunction as a plausible contributor to the chromosomal aberrations found in complex sarcomas. PMID- 15111299 TI - The vascular endothelial growth factor (VEGF) receptor Flt-1 (VEGFR-1) modulates Flk-1 (VEGFR-2) signaling during blood vessel formation. AB - Mice lacking the vascular endothelial growth factor (VEGF) receptor flt-1 (VEGFR 1) die from vascular overgrowth, caused primarily by aberrant endothelial cell division (Kearney JB, Ambler CA, Monaco KA, Johnson N, Rapoport RG, Bautch VL: Vascular endothelial growth factor receptor Flt-1 negatively regulates developmental blood vessel formation by modulating endothelial cell division. Blood 2002, 99:2397-2407). Because a second high-affinity VEGF receptor, flk-1, produces a positive endothelial proliferation signal, it was logical to ask whether flt-1 affects developmental blood vessel formation by modulating signaling through flk-1. Differentiated embryonic stem cell cultures lacking flt 1 (flt-1-/-) had increased flk-1 tyrosine phosphorylation, indicating that flk-1 signaling is up-regulated in the mutant background. The selective flk-1 inhibitor SU5416 partially rescued the flt-1-/- mutant phenotype, and this rescue was accompanied by a decrease in the relative amount of flk-1 tyrosine phosphorylation. Thus reduced flk-1 signal transduction can partially compensate for the lack of flt-1. The flt-1-/- mutant phenotype was also partially rescued by Flt-1/Fc, a truncated flt-1 that binds and sequesters the VEGF ligand. Taken together, these data show that down-regulation of flk-1 signaling by two different strategies partially rescues the developmental vascular overgrowth seen in the absence of flt-1, and they support a model whereby flt-1 modulates the flk 1 signal at an early point in the pathway. PMID- 15111300 TI - Characterization of osteoclasts from patients harboring a G215R mutation in ClC-7 causing autosomal dominant osteopetrosis type II. AB - Autosomal dominant osteopetrosis II (ADOII) is a relatively benign disorder caused by a missense mutation in the ClCN7 gene. In this study, we characterize the osteoclasts from patients with ADOII, caused by a G215R mutation, and investigate the effect on osteoclast function in vitro. Osteoclasts from ADOII patients and healthy age- and sex-matched controls, were used to evaluate osteoclastogenesis, cell fusion, acidification, and resorptive activity. ADOII osteoclasts in vivo have increased number and size. However, in vitro we observed no significant changes in the osteoclast formation rate, the morphology, and the expression of markers, such as cathepsin K and tartrate-resistant acid phosphatase. When mature ADOII osteoclasts were investigated on mineralized bone, they degraded the bone material, however only to 10 to 20% of the level in controls. We show by acridine orange, that the reduced chloride transport leads to reduced acidification. We show that the residual activity is sensitive to inhibitors of cathepsins and chloride channels, confirming that resorption is reduced but present. In conclusion, this is the first functional in vitro study of human ADOII osteoclasts. We show normal osteoclastogenesis in ADOII osteoclasts. However, the residual activity of the ClC-7 channel in ADOII osteoclasts does not allow sufficient acidification and thereby resorption. PMID- 15111302 TI - Neuronal apoptosis is mediated by CXCL10 overexpression in simian human immunodeficiency virus encephalitis. AB - Inflammatory mediators play a crucial role in the pathophysiology of several neurodegenerative diseases including acquired immune deficiency syndrome dementia complex. In the present study we identified a link between CXCL10 overexpression in the brain and human immunodeficiency virus dementia and demonstrated the presence of the chemokine CXCL10 and its receptor, CXCR3, in the neurons in the brains of macaques with simian human immunodeficiency virus encephalitis. Using human fetal brain cultures, we showed that treatment of these cells with either SHIV89.6P or viral gp120 resulted in induction of CXCL10 in neurons. Cultured neurons treated with the chemokine developed increased membrane permeability followed by apoptosis via activation of caspase-3. We confirmed the relevance of these findings in sections of human and macaque brains with encephalopathy demonstrating that neurons expressing CXCL10 also expressed caspase-3. PMID- 15111301 TI - Abrogation of nuclear factor-kappaB activation is involved in zinc inhibition of lipopolysaccharide-induced tumor necrosis factor-alpha production and liver injury. AB - Endotoxin (lipopolysaccharide, LPS)-induced tumor necrosis factor-alpha (TNF alpha) release from Kupffer cells is critically involved in the pathogenesis of alcohol-induced liver injury. We recently reported that inhibition of alcohol induced plasma endotoxin elevation contributes to the protective action of zinc against alcoholic hepatotoxicity. The present study was undertaken to determine whether zinc interferes with the endotoxin-TNF-alpha signaling pathway, and possible mechanism(s) by which zinc modulates the endotoxin-TNF-alpha signaling. Administration of LPS to metallothionein (MT)-knockout (MT-KO) mice and 129/Sv wild-type (WT) controls at 4 mg/kg induced hepatic TNF-alpha elevation at 1.5 hours, followed by liver injury at 3 hours. Zinc pretreatment (two doses at 5 mg/kg) attenuated TNF-alpha production and liver injury in both MT-KO and WT mice, indicating a MT-independent action of zinc. Immunohistochemical detection of the phosphorylation of I-kappaB and nuclear factor (NF)-kappaB in the liver of MT-KO mice demonstrated that zinc pretreatment abrogated LPS-induced NF-kappaB activation in the Kupffer cells. Fluorescent microscopy of superoxide by dihydroethidine and of zinc ions by Zinquin in the liver of MT-KO mice showed that zinc pretreatment increased the intracellular labile zinc ions and inhibited LPS-induced superoxide generation. These results demonstrate that zinc inhibits LPS-induced hepatic TNF-alpha production through abrogation of oxidative stress sensitive NF-kappaB pathway, and the action of zinc is independent of MT. Thus, zinc may be beneficial in the treatment of LPS-induced liver injuries, such as sepsis and alcoholism. PMID- 15111303 TI - Cyclophilin A as a novel biphasic mediator of endothelial activation and dysfunction. AB - Inflammation-mediated endothelial cell (EC) dysfunction likely contributes to the pathogenesis of several vascular diseases including atherosclerosis. We found that stimulation of human umbilical vein ECs with lipopolysaccharide induced secretion of cyclophilin (CyPA) an intracellular protein belonging to the immunophilin family. We then found that when added exogenously CyPA has direct effects on ECs in vitro. At low concentrations (10 to 100 ng/ml) CyPA increased EC proliferation, migration, invasive capacity, and tubulogenesis. Gelatin zymography indicated increased secretion of active matrix metalloproteinase-2, a mediator of cell migration and angiogenesis. At high concentrations (eg, 2 microg/ml) CyPA had opposite effects, decreasing EC migration and viability, possibly in relation to induction of Toll-like receptor-4 expression, detected by immunocytochemistry and flow cytometry. In vivo CyPA expression was not detectable in the luminal ECs of normal mouse carotid arteries but was rapidly induced after systemic lipopolysaccharide injection. In an experimental mouse model of atherosclerosis, CyPA expression was detected in the ECs of neocapillaries of carotid artery lesions, supporting its association with pathological angiogenesis suggested by our in vitro results. In conclusion, we found that CyPA has a biphasic activity on ECs in vitro and is up-regulated in vivo in ECs under pathological states. Our results suggest that CyPA is a novel paracrine and autocrine modulator of EC functions in immune-mediated vascular disease. PMID- 15111304 TI - Role of discoidin domain receptors 1 and 2 in human smooth muscle cell-mediated collagen remodeling: potential implications in atherosclerosis and lymphangioleiomyomatosis. AB - Obstructive diseases of blood vessels and the lung are characterized by degradation and synthesis of new extracellular matrix (ECM) components. Regulated remodeling of the ECM in diseases such as atherosclerosis and lymphangioleiomyomatosis (LAM), both characterized by excessive accumulation of smooth muscle cells (SMCs), is thought to be controlled in part by cell surface receptors for specific ECM components. Discoidin domain receptors (DDR) 1 and 2 represent a family of tyrosine kinase collagen receptors that are activated by fibrillar collagens. To test the hypothesis that DDR may be involved in ECM remodeling by SMCs in vivo, we analyzed DDR expression by reverse transcriptase polymerase chain reaction and immunohistochemistry and demonstrate that both DDR1 and DDR2 are up-regulated in nodules of LAM as compared to normal controls, and are expressed in lesions of atherosclerosis. In vitro, retroviral overexpression of DDR1 or DDR2 in human SMCs cultured on polymerized collagen gels leads to a reduction of collagen expression and induces matrix metalloproteinase (MMP) 1 at both mRNA and protein levels, but only DDR2 enhances MMP2 activation. Moreover, DDR2 overexpression increases SMC-mediated collagen and elastin degradation in vitro. Using laser microdissection, we extend our studies to the analysis of SMCs from LAM nodules where we observe higher MMP1 expression and MMP2 activation. Taken together, these data provide evidence for the potential roles of DDR1 and DDR2 in the regulation of collagen turnover mediated by SMCs in obstructive diseases of blood vessels and the lung. PMID- 15111305 TI - Dynamic populations of dendritic cell-specific ICAM-3 grabbing nonintegrin positive immature dendritic cells and liver/lymph node-specific ICAM-3 grabbing nonintegrin-positive endothelial cells in the outer zones of the paracortex of human lymph nodes. AB - In the paracortex of lymph nodes, cellular immune responses are generated against antigens captured in peripheral tissues by dendritic cells (DCs). DC-SIGN (dendritic cell-specific ICAM-3 grabbing nonintegrin), a C-type lectin exclusively expressed by DCs, functions as an antigen receptor as well as an adhesion receptor. A functional homologue of DC-SIGN, L-SIGN (liver/lymph node SIGN, also called DC-SIGN-related), is expressed by liver sinus endothelial cells. In lymph nodes, both DC-SIGN and L-SIGN are expressed. In this study, we analyzed the distribution of these two SIGN molecules in detail in both normal and immunoreactive lymph nodes. DC-SIGN is expressed by mature DCs in paracortical areas and in addition by DCs with an immature phenotype in the outer zones of the paracortex. L-SIGN expression was also detected in the outer zones on sinus endothelial cells characterized by their expression of the lymphatic endothelial markers LYVE-1 and CLEVER-1. During both cellular and humoral immune responses changes in the amount of DC-SIGN+ immature and mature DCs and L-SIGN+ endothelial cells were observed, indicating that the influx or proliferation of these cells is dynamically regulated. PMID- 15111307 TI - Interaction of Nkx3.1 and p27kip1 in prostate tumor initiation. AB - The homeodomain transcription factor Nkx3.1 and the cyclin-dependent kinase inhibitor p27kip1 have both been implicated in prostate tumor suppression. In addition, both of these molecules demonstrate haploinsufficiency for tumor suppression, in which loss of a single allele is sufficient to lead to the development of preneoplastic or neoplastic lesions. We have generated mice carrying compound mutant alleles of Nkx3.1 and p27 to explore the roles of these factors in prostate tumorigenesis. Our results indicate that Nkx3.1 and p27kip1 cooperate to suppress the proliferation of prostatic epithelial cells and the formation of preneoplastic lesions resembling prostatic intraepithelial neoplasia. Cooperativity was most evident with complete loss of at least one of the two genes because compound heterozygous mice exhibited a prostatic phenotype that was no more severe than that of single heterozygous mutants. Thus Nkx3.1 and p27kip1 regulate prostatic epithelial cell proliferation and tumor initiation by affecting both haploinsufficient and nonhaploinsufficient pathways. PMID- 15111306 TI - Tissue distribution, biochemical properties, and transmission of mouse type A AApoAII amyloid fibrils. AB - In mouse strains with the amyloidogenic apolipoprotein A-II (ApoA-II) gene (Apoa2c), the type C ApoA-II protein (APOAIIC) associates to form amyloid fibrils AApoAII(C) that lead to development of early onset and systemic amyloidosis with characteristic heavy amyloid deposits in the liver and spleen. We found age associated heavy deposition of amyloid fibrils [AApoAII(A)] composed of type A ApoA-II protein (APOAIIA) in BDF1 and C57BL/6 mice reared at one of our institutes. AApoAII(A) fibrils were deposited in the intestine, lungs, tongue, and stomach but not in the liver or spleen. AApoAII(A) fibrils were isolated, and morphological, biochemical, and structural characteristics distinct from those seen in AApoAII(C) and mouse AA amyloid fibrils were found. Transmission electron and atomic force microscopy showed that the majority of isolated AApoAII(A) amyloid fibrils featured fine, protofibril-like shapes. AApoAII(A) fibrils have a much weaker affinity for thioflavine T than for AApoAII(C), whereas APOAIIA protein contains less of the beta-pleated sheet structure than does APOAIIC. The injection of AApoAII(A) fibrils induced amyloid deposition in C57BL/6 and DBA2 mice (Apoa2a) as well as in R1.P1-Apoa2c mice (Apoa2c), but AApoAII(A) induced more severe amyloidosis in Apoa2a strains than in the Apoa2c strain. It was found that AApoAII(A) fibrils isolated from mice with mildly amyloidogenic APOAIIA protein have distinct characteristics. Induction of amyloidosis by heterologous amyloid fibrils clearly showed interactions between amyloid protein monomers and fibrils having different primary structures. PMID- 15111309 TI - Clostridium perfringens enterotoxin elicits rapid and specific cytolysis of breast carcinoma cells mediated through tight junction proteins claudin 3 and 4. AB - Clostridium perfringens enterotoxin (CPE) induces cytolysis very rapidly through binding to its receptors, the tight junction proteins CLDN 3 and 4. In this study, we investigated CLDN 3 and 4 expression in breast cancer and tested the potential of CPE-mediated therapy. CLDN 3 and 4 proteins were detected in all primary breast carcinomas tested (n = 21) and, compared to normal mammary epithelium, were overexpressed in approximately 62% and 26%, respectively. Treatment of breast cancer cell lines in culture with CPE resulted in rapid and dose-dependent cytolysis exclusively in cells that expressed CLDN 3 and 4. Intratumoral CPE treatment of xenografts of T47D breast cancer cells in immunodeficient mice resulted in a significant reduction in tumor volume (P = 0.007), with accompanying necrosis. Necrotic reactions were also seen in three freshly resected primary breast carcinoma samples treated with CPE for 12 hours, while isolated primary breast carcinoma cells underwent rapid and complete cytolysis within 1 hour. Thus, expression of CLDN 3 and 4 sensitizes primary breast carcinomas to CPE-mediated cytolysis and emphasizes the potential of CPE in breast cancer therapy. PMID- 15111308 TI - Neural stem/progenitor cells express costimulatory molecules that are differentially regulated by inflammatory and apoptotic stimuli. AB - Increased expression of the costimulatory molecule CD80 (B7-1) was noted in the subventricular zone of the brain during the course of experimental autoimmune encephalomyelitis (EAE). This area of the brain is a neural stem cell (NSC) niche in the adult. We show that isolated NSCs from adult brain express CD80 and CD86 (B7-2) and this expression is increased after exposure to IFN-gamma or TNF-alpha, the prototypical Th1 cytokines expressed during EAE. CD80 and CD86 expressed by NSCs are functional and can costimulate allogeneic cells in a mixed lymphocyte reaction. Furthermore, cross-linking of CD80 on the surface of NSCs results in apoptosis of NSCs. In vitro, we show that T cells can interact with NSCs and form conjugates with redistribution of CD3 on the surface of T cells to the area of contact. These data raise the possibility that during CNS inflammatory diseases such as EAE, NSCs may express immune molecules and interact with the inflammatory environment potentially resulting in injury to the NSCs, which may have implications for repair mechanisms in the central nervous system. PMID- 15111310 TI - Detection of a sulfotransferase (HEC-GlcNAc6ST) in high endothelial venules of lymph nodes and in high endothelial venule-like vessels within ectopic lymphoid aggregates: relationship to the MECA-79 epitope. AB - The interaction of L-selectin on lymphocytes with sulfated ligands on high endothelial venules (HEVs) of lymph nodes results in lymphocyte rolling and is essential for lymphocyte homing. The MECA-79 monoclonal antibody reports HEV expressed ligands for L-selectin by recognizing a critical sulfation-dependent determinant on these ligands. HEC-GlcNAc6ST, a HEV-localized sulfotransferase, is essential for the elaboration of functional ligands within lymph nodes, as well as the generation of the MECA-79 epitope. Here, we use an antibody against murine HEC-GlcNAc6ST to study its expression in relationship to the MECA-79 epitope. In lymph nodes, the enzyme is expressed in the Golgi apparatus of high endothelial cells, in close correspondence with luminal staining by MECA-79. In lymph node HEVs of HEC-GlcNAc6ST-null mice, luminal staining by MECA-79 is almost abolished, whereas abluminal staining persists although reduced in intensity. HEV-like vessels in several examples of inflammation-associated lymphoid neogenesis, including nonobese diabetic mice, also exhibit concomitant expression of the sulfotransferase and luminal MECA-79 reactivity. The correlation extends to ectopic lymphoid aggregates within the pancreas of RIP-BLC mice, in which CXCL13 is expressed in islets. Analysis of the progeny of RIP-BLC by HEC-GlcNAc6ST-null mice establishes that the enzyme is responsible for the MECA-79 defined luminal ligands. PMID- 15111311 TI - Activation of the GLI oncogene through fusion with the beta-actin gene (ACTB) in a group of distinctive pericytic neoplasms: pericytoma with t(7;12). AB - Activation of the GLI oncogene is an important step in the sonic hedgehog signaling pathway, and leads to, eg, tissue-specific cell proliferation during embryogenesis. GLI activity in adult tissues is restricted, but has been identified in various neoplasms, as a result of mutations in the PTCH (patched) or SMOH (smoothened) genes, encoding components of the sonic hedgehog pathway, or by amplification of GLI. Herein, we present a new mechanism of GLI activation through fusion with the beta-actin gene (ACTB) in five histologically distinctive soft tissue tumors showing a t(7;12)(p21-22;q13-15) and a pericytic phenotype. Each was composed of a perivascular proliferation of monomorphic short spindle cells that stained positively for smooth muscle actin and laminin and that showed pericytic features by electron microscopy. To date, with a median follow-up of 24 months, none has behaved in an aggressive manner. Molecular genetic analysis showed that the translocation in all cases resulted in a fusion transcript including the 5'-part of ACTB and the 3'-part of GLI. The DNA-binding zinc finger domains of GLI were retained in the fusion transcripts and it is likely that the replacement of the promoter region of GLI with that of the ubiquitously expressed ACTB gene leads to deregulation of GLI expression and its downstream target genes. PMID- 15111312 TI - 12/15-lipoxygenase is increased in Alzheimer's disease: possible involvement in brain oxidative stress. AB - Alzheimer's disease (AD) is a chronic neurodegenerative disorder that impairs cognition and behavior. Although the initiating molecular events are not known, increasing evidence suggests that oxidative stress could play a functional role in its pathogenesis. Lipoxygenase (LOX) enzymes by oxidizing polyunsaturated fatty acids synthesize hydroperoxyacids, which are potent pro-oxidant mediators. Because circumstantial evidence suggests that 12/15-LOX is a major source of oxidative stress, we investigated the protein levels and activity of this enzyme in different brain regions of histopathologically confirmed AD and control cases. Using quantitative Western blot analysis we demonstrated that in affected frontal and temporal regions of AD brains the amount of 12/15-LOX was higher compared with controls, whereas no difference between the two groups was detected in the cerebellum. This observation was confirmed by immunohistochemical studies. Levels of 12/15-hydroxyeicosatetraenoic acids, metabolic products of 12/15-LOX, were also markedly elevated in AD brains compared to controls. This increase directly correlated with brain lipid peroxidation, and inversely with vitamin E levels. Finally, genetic deletion of this enzyme in vitro resulted in a reduction of the cellular oxidative stress response after incubation with H2O2 or amyloid beta. These data show that the 12/15-LOX metabolic pathway is increased and correlates with an oxidative imbalance in the AD brain, implying that this enzyme might contribute to the pathogenesis of this neurodegenerative disorder. PMID- 15111313 TI - Tumor necrosis factor-alpha induces fractalkine expression preferentially in arterial endothelial cells and mithramycin A suppresses TNF-alpha-induced fractalkine expression. AB - Fractalkine is an unusual tumor necrosis factor (TNF)-alpha-induced chemokine. The molecule is tethered to cells that express it and produces strong and direct adhesion to leukocytes expressing fractalkine receptor. However, the potential mechanism and significance of TNF-alpha-induced fractalkine expression in vascular endothelial cells are poorly understood. Here we show that in primary cultured endothelial cells TNF-alpha-induced fractalkine mRNA expression is mediated mainly through phosphatidylinositol 3'-kinase activation and nuclear factor (NF)-kappaB mediated transcriptional activation, along with GC-rich DNA binding protein-mediated transcription. Interestingly, GC-rich DNA-binding protein inhibitors, mithramycin A and chromomycin A3, strongly suppressed TNF alpha-induced fractalkine mRNA expression, possibly through inhibition of transcriptional activities by NF-kappaB and Sp1. In fact, direct inhibition of NF kappaB and Sp1 bindings by decoy oligonucleotides suppressed TNF-alpha-induced fractalkine expression. Histologically, TNF-alpha-induced fractalkine expression was observed markedly in arterial and capillary endothelial cells, endocardium, and endothelium of intestinal villi, and slightly in venous endothelial cells, but not at all in lymphatic endothelial cells of intestine. Mithramycin A markedly suppressed TNF-alpha-induced fractalkine expression in vivo. These results indicate that TNF-alpha-stimulated fractalkine expression could act as part of arterial endothelial adhesion to leukocytes and monocytes during inflammation and atherosclerosis. NF-kappaB and Sp1 inhibitors such as mithramycin A may provide a pharmacological approach to suppressing these processes. PMID- 15111314 TI - Oligodendrocytes and progenitors become progressively depleted within chronically demyelinated lesions. AB - To understand mechanisms that may underlie the progression of a demyelinated lesion to a chronic state, we have used the cuprizone model of chronic demyelination. In this study, we investigated the fate of oligodendrocytes during the progression of a demyelinating lesion to a chronic state and determined whether transplanted adult oligodendrocyte progenitors could remyelinate the chronically demyelinated axons. Although there is rapid regeneration of the oligodendrocyte population following an acute lesion, most of these newly regenerated cells undergo apoptosis if mice remain on a cuprizone diet. Furthermore, the oligodendrocyte progenitors also become progressively depleted within the lesion, which appears to contribute to the chronic demyelination. Interestingly, even if the mice are returned to a normal diet following 12 weeks of exposure to cuprizone, remyelination and oligodendrocyte regeneration does not occur. However, if adult O4+ progenitors are transplanted into the chronically demyelinated lesion of mice treated with cuprizone for 12 weeks, mature oligodendrocyte regeneration and remyelination occurs after the mice are returned to a normal diet. Thus, the formation of chronically demyelinated lesions induced by cuprizone appears to be the result of oligodendrocyte depletion within the lesion and not due to the inability of the chronically demyelinated axons to be remyelinated. PMID- 15111315 TI - N-acetyl-cysteine promotes angiostatin production and vascular collapse in an orthotopic model of breast cancer. AB - The antioxidant N-acetyl-cysteine (NAC) has been shown to be chemopreventive in clinical studies, and in recent studies, has shown promise in preventing tumor progression. Although the effects of NAC on tumorigenesis have been associated with decreased angiogenesis, the mechanism of the anti-angiogenic activity has not been determined. In the following study, we describe a novel mechanism whereby NAC therapy blocks MDA-MB-435 breast carcinoma cell proliferation and metastasis in an in vivo tumorigenic model. Athymic nude mice bearing MDA-MB-435 xenografts were treated with systemic NAC daily for 8 weeks. NAC treatment resulted in endothelial cell apoptosis and reduction of microvascular density within the core of the tumor leading to significant tumor cell apoptosis/necrosis. Angiostatin accumulated in tumors from NAC-treated but not control animals. Additional studies using a vascular endothelial growth factor dependent chicken chorioallantoic membrane angiogenic assay recapitulated NAC induced endothelial apoptosis and coordinate production of angiostatin, a potent endothelial apoptotic factor. In vitro studies showed angiostatin was formed in endothelial cultures in a vascular endothelial growth factor- and NAC-dependent manner, a process that requires endothelial cell surface plasminogen activation. These results suggest that systemic NAC therapy promotes anti-angiogenesis through angiostatin production, resulting in endothelial apoptosis and vascular collapse in the tumor. PMID- 15111316 TI - Simvastatin inhibits leukocyte accumulation and vascular permeability in the retinas of rats with streptozotocin-induced diabetes. AB - Leukocytes play important roles in the pathogenesis of diabetic retinopathy. Recently, 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors have been reported to exert various effects in addition to their lipid-lowering ability. We investigated the effects of simvastatin, a 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor, on leukocyte-induced diabetic changes in retinas. Diabetes was induced in Long-Evans rats with streptozotocin, and simvastatin administration was begun immediately after the induction of diabetes. Two weeks of treatment with simvastatin suppressed significantly the number of leukocytes adhering to retinal vessel endothelium and the number of leukocytes accumulated in the retinal tissue by 72.9% and 41.0%, respectively (P < 0.01). The expression of intercellular adhesion molecule-1 (ICAM-1) and the CD18 (the common beta-chain of ICAM-1 ligands) were both suppressed with simvastatin. The amount of vascular endothelial growth factor in the retina was attenuated in the simvastatin-treated group. To evaluate the effects of simvastatin on leukocyte-induced endothelial cell damage, vascular permeability in the retina was measured with fluorescein labeled dextran. Treatment with simvastatin markedly reduced retinal permeability (P = 0.014). This suggests that simvastatin attenuates leukocyte-endothelial cell interactions and subsequent blood-retinal barrier breakdown via suppression of vascular endothelial growth factor-induced ICAM-1 expression in the diabetic retina. Simvastatin may thus be useful in the prevention of diabetic retinopathy. PMID- 15111317 TI - Pancreatic trypsin increases matrix metalloproteinase-9 accumulation and activation during acute intestinal ischemia-reperfusion in the rat. AB - Ischemia-reperfusion of the intestine produces a set of inflammatory mediators, the origin of which has recently been shown to involve pancreatic digestive enzymes. Matrix metalloproteinase-9 (MMP-9) participates in a variety of inflammatory processes including myocardial, hepatic, and pancreatic ischemia reperfusion. In the present study, we explore the role of neutrophil-derived MMP 9 in acute intestinal ischemia-reperfusion and its interaction with pancreatic trypsin. Male Sprague-Dawley rats were subjected to 45 minutes of superior mesenteric arterial occlusion followed by 90 minutes of reperfusion. In situ zymography of the proximal jejunum reveals increased gelatinase activity in the intestinal wall after ischemia-reperfusion. Gel electrophoresis zymography and immunofluorescence co-localization suggests that this gelatinase activity is derived from MMP-9 released from infiltrating neutrophils. The role of intraluminal trypsin in this process was investigated using an in vivo isolated jejunal loop model of intestinal ischemia-reperfusion. Trypsin increased the inflammatory response after reperfusion, with an augmented neutrophil infiltration of the intestinal wall. Furthermore, trypsin stimulated a rapid conversion of neutrophil-released proMMP-9 into the lower molecular weight enzymatically active MMP-9. This process represents a powerful in vivo pathophysiological mechanism for trypsin-induced MMP-9 activation and is likely to play a central role in the development of acute intestinal inflammation and shock. PMID- 15111318 TI - Anti-human Olig2 antibody as a useful immunohistochemical marker of normal oligodendrocytes and gliomas. AB - Olig2 is a recently identified transcription factor involved in the phenotype definition of cells in the oligodendroglial lineage. The expression of Olig2 transcript has been demonstrated in human oligodendroglial tumors, although the protein expression has not been studied extensively. We developed a polyclonal antibody to human Olig2 and analyzed it immunohistochemically. The antibody depicted a single distinct band of predicted molecular weight by Western blotting, and did not cross-react with human Olig1. In normal human brain tissue, the nuclei of oligodendrocytes of interfascicular, perivascular, and perineuronal disposition were clearly labeled by the antibody. Similarly, the nuclei of oligodendroglial tumors were labeled. There was no apparent correlation between the staining intensity and histological grade. Astrocytic components within the tumors were generally less or not stained. Astrocytic tumors were also positive with the Olig2 antiserum to a lesser extent, and the difference between oligodendroglial and astrocytic tumors was demonstrated by a statistical analysis. Olig2 and glial fibrillary acidic protein were expressed in a mutually exclusive manner, and Olig2 expression was cell-cycle related. Neither central neurocytoma nor schwannoma cases were stained. Our antibody was demonstrated to be useful in recognizing normal oligodendrocytes on paraffin sections, and applicable in diagnosis of some brain tumors. PMID- 15111319 TI - Prevalent overexpression of prolyl isomerase Pin1 in human cancers. AB - Phosphorylation of proteins on serine or threonine residues preceding proline (pSer/Thr-Pro) is a major regulatory mechanism in cell proliferation and transformation. Interestingly, the pSer/Thr-Pro motifs in proteins exist in two distinct cis and trans conformations, whose conversion rate is normally reduced on phosphorylation, but is catalyzed specifically by the prolyl isomerase Pin1. Pin1 can catalytically induce conformational changes in proteins after phosphorylation, thereby having profound effects on catalytic activity, dephosphorylation, protein-protein interactions, subcellular location, and/or turnover of certain phosphorylated proteins. Recently, it has been shown that Pin1 is overexpressed in human breast cancer cell lines and cancer tissues and plays a critical role in the transformation of mammary epithelial cells by activating multiple oncogenic pathways. Furthermore, Pin1 expression is an excellent independent prognostic marker in prostate cancer. However, little is known about Pin1 expression in other human normal and cancerous tissues. In the present study, we quantified Pin1 expression in 2041 human tumor samples and 609 normal tissue samples as well as normal and transformed human cell lines. We found that Pin1 was usually expressed at very low levels in most normal tissues and its expression was normally associated with cell proliferation, with high Pin1 levels being found only in a few cell types. However, Pin1 was strikingly overexpressed in many different human cancers. Most tumors (38 of 60 tumor types) have Pin1 overexpression in more than 10% of the cases, as compared with the corresponding normal controls, which included prostate, lung, ovary, cervical, brain tumors, and melanoma. Consistent with these findings, Pin1 expression in human cancer cell lines was also higher than that in the normal cell lines examined. These results indicate that Pin1 overexpression is a prevalent and specific event in human cancers. Given previous findings that Pin1 expression is an excellent prognostic marker in prostate cancer and that inhibition of Pin1 can suppress transformed phenotypes and inhibit tumor cell growth, these findings may have important implications for the pathogenesis, diagnosis, and treatment of human cancers. PMID- 15111320 TI - Beta-catenin simultaneously induces activation of the p53-p21WAF1 pathway and overexpression of cyclin D1 during squamous differentiation of endometrial carcinoma cells. AB - The functional consequences of up-regulation of beta-catenin as a transcription factor are complex in different tumors. To clarify roles during squamous differentiation (SqD) of endometrial carcinoma (Em Ca) cells, we investigated expression of beta-catenin, as well as cyclin D1, p53, p21WAF1, and PML (promyelocytic leukemia) in 80 cases of Em Ca with SqD areas, in comparison with cell proliferation determined with reference to Ki-67 antigen positivity. The impact of beta-catenin-T-cell factor (TCF)-mediated transcription was also examined using Em Ca cells. In clinical cases, nuclear beta-catenin accumulation was more frequent in SqD areas, being positively linked with expression of cyclin D1, p53, and p21WAF1, and inversely with Ki-67 and PML immunoreactivity. Significant correlations of nuclear beta-catenin, cyclin D1, p53, and p21WAF1 were noted between SqD and the surrounding carcinoma lesions. The Ishikawa cell line, with stable or tetracycline-regulated expression of mutant beta-catenin, showed an increase in expression levels of cyclin D1, p14ARF, p53, and p21WAF1 but not PML, and activation of beta-catenin-TCF4-mediated transcription determined with TOP/FOP constructs. The cell morphology was senescence-like rather than squamoid in appearance. Moreover, overexpressed beta-catenin could activate transcription from p14ARF and cyclin D1 promoters, in a TCF4-dependent manner. These findings indicate that in Em Cas, nuclear beta-catenin can simultaneously induce activation of the p53-p21WAF1 pathway and overexpression of cyclin D1, leading to suppression of cell proliferation or induction of cell senescence. However, overexpression of beta-catenin alone is not sufficient for development of a squamoid phenotype in Em Ca cells, suggesting that nuclear accumulation is an initial signal for trans-differentiation. PMID- 15111321 TI - The effect of dexamethasone on the generation of plasma DNA from dead and dying cells. AB - To determine the effects of glucocorticoids on the clearance of apoptotic and necrotic cells, the influence of dexamethasone on plasma levels of DNA was assessed in BALB/c mice receiving Jurkat cells treated with etoposide or ethanol. In untreated mice, administration of 10(8) apoptotic or necrotic Jurkat cells led to the appearance of DNA in the plasma. In mice treated 24 hours previously with dexamethasone, levels of DNA were reduced in a dose-dependent manner, with mice receiving 1 and 2.5 mg showing no appreciable plasma DNA levels. Similar results were obtained with assay of lactate dehydrogenase in mice receiving apoptotic cells. The effects of dexamethasone on anti-Fas treatment were also characterized. While treatment with a monoclonal anti-Fas reagent caused a significant plasma DNA response in untreated mice, mice pretreated with dexamethasone showed much lower levels. Blood levels of caspase 3 and TUNEL staining of liver were also reduced in dexamethasone-treated mice compared to controls receiving anti-Fas antibody. These results indicate that glucocorticoids can affect the clearance of apoptotic and necrotic cells as well as the induction of apoptosis in at least some tissues. These activities may be relevant to the efficacy of glucocorticoids in the treatment of inflammatory disease. PMID- 15111322 TI - Urease expression by Cryptococcus neoformans promotes microvascular sequestration, thereby enhancing central nervous system invasion. AB - Our objective was to determine the role of the cryptococcal virulence factor urease in pulmonary-to-central nervous system, dissemination, invasion, and growth. C. neoformans H99, the urease knockout strain (ure1) derived from H99, and the urease restored strain ure1+URE1-1 were used for the studies. The absence of cryptococcal urease (ure1infection) resulted in significant protection from the high mortality observed in H99-infected mice. All H99-infected mice had extremely high cryptococcal loads in their brains at the time of death, whereas only two of six animals that died of ure1 infection had detectable C. neoformans in the brain. Histological analysis of the blood-to-brain invasion by C. neoformans H99 demonstrated wedging of the yeasts in small capillaries, altered structure of microvessel walls, formation of mucoid cysts initiated in the proximity of damaged microcapillaries, and the absence of an inflammatory response. Direct inoculation of H99, ure1, and ure1+URE1-1 into the brain demonstrated that urease was not required to grow in the brain. However, the dissemination patterns in the brain, spleen, and other organs after intravenous inoculation indicated that cryptococcal urease contributes to the central nervous system invasion by enhancing yeast sequestration within microcapillary beds (such as within the brain) during hematogenous spread, thereby facilitating blood-to brain invasion by C. neoformans. PMID- 15111323 TI - Cloning of cDNA encoding a regeneration-associated muscle protease whose expression is attenuated in cell lines derived from Duchenne muscular dystrophy patients. AB - In the dystrophin-mutant mdx mouse, an animal model for Duchenne muscular dystrophy (DMD), damaged skeletal muscles are efficiently regenerated and thus the animals thrive. The phenotypic differences between DMD patients and the mdx mice suggest the existence of factors that modulate the muscle wasting in the mdx mice. To identify these factors, we searched for mRNAs affected by the mdx mutation by using cDNA microarrays with newly established skeletal muscle cell lines from mdx and normal mice. We found that in the mdx muscle cell line, 12 genes, including L-arginine:glycine amidinotransferase and thymosin beta4, are up regulated, whereas 7 genes, including selenoprotein P and a novel regeneration associated muscle protease (RAMP), are down-regulated. Northern blot analysis and in situ hybridization revealed that RAMP mRNA is predominantly expressed in normal skeletal muscle and brain, and its production is enhanced in the regenerating area of injured skeletal muscle in mice. RAMP expression was much lower in individual muscle cell lines derived from biopsies of six DMD patients compared to a normal muscle cell line. These results suggest that RAMP may play a role in the regeneration of skeletal muscle and that its down-regulation could be involved in the progression of DMD in humans. PMID- 15111324 TI - Mechanisms of cell-cycle arrest in Spitz nevi with constitutive activation of the MAP-kinase pathway. AB - Spitz nevi are benign melanocytic nevi that overlap histopathologically with melanoma. We previously found copy number increases of chromosome 11p frequently paralleled by mutations in the HRAS oncogene mapping to this region. In this study, we explored mechanisms that inhibit proliferation in the presence of HRAS activation. We analyzed MAP-kinase activation using immunohistochemistry for phospho-ERK, cyclin D1, and microphthalmia transcription factor expression in 17 Spitz nevi with and 18 Spitz nevi without 11p copy number increase. We found relatively high levels of phospho-ERK and cyclin D1 expression suggesting MAP kinase pathway activation in both groups of Spitz nevi. However, Spitz nevi with 11p copy number increases showed significantly higher levels of cyclin D1 expression and lower levels of microphthalmia transcription factor expression suggesting stronger MAP-kinase pathway activation in this group. Contrasting this apparent activation, the proliferation rate as assessed by Mib1 expression was low in both groups. An analysis of cell-cycle inhibitory proteins including p16, p21, and p27 showed that the majority of Spitz nevus cells expressed high levels of p16, with cells of the cases that had increased copy number of 11p expressing significantly higher levels than those of Spitz nevi with normal copy number of 11p. We propose that in benign nevi with constitutive activation of the MAP kinase pathway, p16 functions as an essential mediator of oncogene-induced senescence preventing progression to melanoma. PMID- 15111325 TI - Activation of peroxisome proliferator-activated receptor-gamma reverses squamous metaplasia and induces transitional differentiation in normal human urothelial cells. AB - We observed that in urothelium, both cornifying and noncornifying forms of squamous metaplasia are accompanied by changes in the localization of the nuclear hormone receptors, peroxisome proliferator activated receptor gamma (PPAR-gamma) and retinoid X receptor (RXR-alpha). To obtain objective evidence for a role for PPAR-gamma-mediated signaling in urothelial differentiation, we examined expression of the cytokeratin isotypes CK13, CK20, and CK14 as indicators of transitional, terminal transitional, and squamous differentiation, respectively, in cultures of normal human urothelial cells. In control culture conditions, normal human urothelial cells showed evidence of squamous differentiation (CK14+, CK13-, CK20-). Treatment with the high-affinity PPAR-gamma agonist, troglitazone (TZ), resulted in gain of CK13 and loss of CK14 protein expression. The effect of TZ was significantly augmented when the autocrine-stimulated epidermal growth factor receptor pathway was inhibited and this resulted in induction of CK20 expression. The RXR-specific inhibitors PA452, HX531, and HX603 inhibited the TZ induced CK13 expression, supporting a role for RXR in the induction of CK13 expression. Thus, signaling through PPAR-gamma can mediate transitional differentiation of urothelial cells and this is modulated by growth regulatory programs. PMID- 15111326 TI - Expression of myopodin induces suppression of tumor growth and metastasis. AB - Myopodin was previously reported as a gene that was frequently deleted in prostate cancer. This gene shares significant homology with a cell shape regulating gene, synaptopodin. Myopodin was shown to bind actin and to induce actin bundling when cells were stimulated. To clarify the functional role of myopodin in prostate cancer, several assays were performed to evaluate the tumor suppression activity of myopodin. Our results indicate that myopodin inhibits tumor growth and invasion both in vitro and in vivo. The activity of tumor suppression of myopodin is located at the C-terminus region. To further evaluate the role of myopodin in suppressing the invasiveness of prostate cancer, an expression analysis of myopodin protein was performed in prostate tissues. The results indicate that down-regulation of myopodin expression occurs mostly in invasive stages of prostate cancer, implying a potential invasion suppression role for myopodin in prostate cancer. In addition, hemizygous deletion and down regulation of myopodin expression occur in three aggressive prostate cancer cell lines. All these results support the hypothesis that myopodin functions as a tumor suppressor gene to limit the growth and to inhibit the metastasis of cancer cells. PMID- 15111327 TI - Proteomic analysis of exosomes secreted by human mesothelioma cells. AB - Exosomes are small membrane vesicles secreted into the extracellular compartment by exocytosis. Tumor exosomes may be involved in the sampling of antigens to antigen presenting cells or as decoys allowing the tumor to escape immune directed destruction. The proteins present in exosomes secreted by tumor cells have been poorly defined. This study describes the protein composition of mesothelioma cell-derived exosomes in more detail. After electrophoresis of exosome preparations, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) was used to characterize the protein spots. MHC class I was found to be present together with the heat shock proteins HSC70 and HSP90. In addition, we found annexins and PV-1, proteins involved in membrane transport and function. Cytoskeleton proteins and their associated proteins ezrin, moesin, actinin-4, desmoplakin, and fascin were also detected. Besides the molecular motor kinesin like protein, many enzymes were detected revealing the cytoplasmic orientation of exosomes. Most interesting was the detection of developmental endothelial locus-1 (DEL-1), which can act as a strong angiogenic factor and can increase the vascular development in the neighborhood of the tumor. In conclusion, mesothelioma cells release exosomes that express a discrete set of proteins involved in antigen presentation, signal transduction, migration, and adhesion. Exosomes may play an important role in the interaction between tumor cells and their environment. PMID- 15111328 TI - Persistent proteinuria up-regulates angiotensin II type 2 receptor and induces apoptosis in proximal tubular cells. AB - Apoptosis is implicated in the progressive cell loss and fibrosis both at glomerular and tubulointerstitial level. In this study, we examined the potential mechanisms by which persistent proteinuria (protein-overload model) could induce apoptosis. After uninephrectomy (UNX), Wistar rats received daily injections of 0.5 g of bovine serum albumin (BSA)/100 g body weight or saline. Both at day 8 and day 28, rats receiving BSA had proteinuria and renal lesions characterized by tubular atrophy and/or dilation and mononuclear cell infiltration. In relation to control-UNX rats, renal cortex of nephritic rats showed an increment in AT2 mRNA (reverse transcriptase-polymerase chain reaction) and protein (Western blot) expression. In both groups, AT2 receptor immunostaining was mainly localized in proximal tubular cells. Rats with persistent proteinuria showed a significantly increased number of terminal dUTP nick-end labeling positive apoptotic cells compared with UNX-controls, both in glomeruli and tubulointerstitium. Double staining for apoptosis and AT2 receptor showed that most terminal dUTP nick-end labeling positive cells were found in tubules expressing AT2 receptor. Using an antibody that recognizes the active form caspase-3, we observed an increment in caspase-3 activation in rats receiving BSA with respect to those receiving saline. Rats with persistent proteinuria showed a diminution in the phosphorylation of Bcl-2 with respect to UNX-controls both at day 8 and day 28. By contrast, no changes were observed either in the Bax or in the Bcl-2 protein levels. The administration of BSA to UNX rats induced a diminution in the phosphorylation of ERK with respect to UNX-control at all times studied. The changes observed in ERK activities took place without alterations of ERK1/2 protein levels. In summary, our data suggest that persistent proteinuria causes apoptosis in tubular cells through the activation of AT2 receptor, which can, in turn, inhibit MAP kinase (ERK1/2) activation and Bcl-2 phosphorylation. PMID- 15111330 TI - Genomic profiling of peripheral T-cell lymphoma, unspecified, and anaplastic large T-cell lymphoma delineates novel recurrent chromosomal alterations. AB - To characterize genetic alterations in peripheral T-cell lymphoma, not otherwise specified (PTCL NOS), and anaplastic large T-cell lymphoma (ALCL), 42 PTCL NOS and 37 ALCL [17 anaplastic large cell kinase (ALK)-negative ALCL, 9 ALK-positive ALCL, 11 cutaneous ALCL] were analyzed by comparative genomic hybridization. Among 36 de novo PTCL NOS, recurrent chromosomal losses were found on chromosomes 13q (minimally overlapping region 13q21, 36% of cases), 6q and 9p (6q21 and 9p21 pter, in 31% of cases each), 10q and 12q (10q23-24 and 12q21-q22, in 28% of cases each), and 5q (5q21, 25% of cases). Recurrent gains were found on chromosome 7q22 qter (31% of cases). In 11 PTCL NOS, high-level amplifications were observed, among them 3 cases with amplification of 12p13 that was restricted to cytotoxic PTCL NOS. Whereas cutaneous ALCL and ALK-positive ALCL showed few recurrent chromosomal imbalances, ALK-negative ALCL displayed recurrent chromosomal gains of 1q (1q41-qter, 46%), and losses of 6q (6q21, 31%) and 13q (13q21-q22, 23%). Losses of chromosomes 5q, 10q, and 12q characterized a group of noncytotoxic nodal CD5+ peripheral T-cell lymphomas. The genetics of PTCL NOS and ALK-negative ALCL differ from other T-NHLs characterized genetically so far, among them enteropathy-type T-cell lymphoma, T-cell prolymphocytic leukemia, and adult T cell lymphoma/leukemia. PMID- 15111329 TI - Transcription factor Ets-1 mediates ischemia- and vascular endothelial growth factor-dependent retinal neovascularization. AB - Transcription factor Ets-1 has been reported to regulate angiogenesis in vascular endothelial cells. Here, we investigated a mechanism that may regulate the expression of Ets-1 in vascular endothelial growth factor (VEGF)- and hypoxia induced retinal neovascularization and that may have potential to inhibit ocular neovascular diseases. VEGF and hypoxia increased Ets-1 expression in cultured bovine retinal endothelial cells. The VEGF-induced mRNA increase of Ets-1 was suppressed by a tyrosine kinase inhibitor (genistein), by inhibitors of MEK (mitogen-activated protein and extracellular signal-regulated kinase kinase) (PD98059 and UO126), and by inhibitors of protein kinase C (GF109203X, staurosporine, and Go6976). Dominant-negative Ets-1 inhibited VEGF-induced cell proliferation, tube formation, and the expression of neuropilin-1 and angiopoietin-2. In a mouse model of proliferative retinopathy, Ets-1 mRNA was up regulated. Intravitreal injection of dominant-negative Ets-1 suppressed retinal angiogenesis in a mouse model of proliferative retinopathy. In conclusion, VEGF induces Ets-1 expression in bovine retinal endothelial cells and its expression is protein kinase C/ERK pathway-dependent. Ets-1 up-regulation is involved in the development of retinal neovascularization, and inhibition of Ets-1 may be beneficial in the treatment of ischemic ocular diseases. PMID- 15111331 TI - Electrophysiological properties of the airway: epithelium in the murine, ovalbumin model of allergic airway disease. AB - The electrophysiological properties of cultured tracheal cells (CTCs) were examined in a murine (C57BL/6J), ovalbumin (OVA)-induced model of allergic airway disease (AAD) at early (3-day OVA-aerosol) and peak (10-day OVA-aerosol) periods of inflammation. Transepithelial potential difference, short-circuit current (Isc), and resistance (RT) were lower in CTCs from 10-day OVA-aerosol animals compared to CTCs from naive mice. In cells cultured for 5 weeks, RT was greater in naive CTCs than in 10-day OVA-aerosol CTCs at all times (P < 0.01). The Isc response to mucosal amiloride (10(-4) mol/L) was increased in CTCs from 10-day OVA-aerosol mice compared to naive mice (6.0 +/- 0.37 microA/cm2 versus 1.8 +/- 0.56 microA/cm2; P < 0.001) with intermediate values for CTCs from 3-day OVA aerosol mice. The cAMP-induced increase in Isc was blunted in 10-day OVA-aerosol animals compared to CTCs from naive mice (9 +/- 12% versus 39 +/- 7%; P < 0.01) with intermediate values for CTCs from 3-day OVA-aerosol mice. There was no difference in mannitol flux in naive compared to 10-day OVA-aerosol CTCs. Similar results were found using intact tracheas mounted in a perfusion chamber. These data demonstrate changes in airway epithelial cell function in the OVA-induced model of AAD that may contribute to the pathogenesis of airway inflammation. PMID- 15111332 TI - The powerful neuroprotective action of C1-inhibitor on brain ischemia-reperfusion injury does not require C1q. AB - C1-inhibitor (C1-INH) is a major regulator of the complement classical pathway. Besides this action, it may also inhibit other related inflammatory systems. We have studied the effect of C1-INH in C57BL/6 mice with focal transient brain ischemia induced by 30 minutes of occlusion of the middle cerebral artery. C1-INH induced a dose-dependent reduction of ischemic volume that, with the dose of 15 U/mouse, reached 10.8% of the volume of saline-treated mice. Four days after ischemia the treated mice had significantly lower general and focal neurological deficit scores. Fluoro-Jade staining, a marker for neuronal degeneration, showed that C1-INH-treated mice had a lower number of degenerating cells. Leukocyte infiltration, as assessed by CD45 immunostaining, was also markedly decreased. We then investigated the response to ischemia in C1q(-/-) mice. There was a slight, nonsignificant decrease in infarct volume in C1q(-/-) mice (reduction to 72.3%) compared to wild types. Administration of C1-INH to these mice was still able to reduce the ischemic volume to 31.4%. The study shows that C1-INH has a strong neuroprotective effect on brain ischemia/reperfusion injury and that its action is independent from C1q-mediated activation of classical pathway. PMID- 15111333 TI - Genetic deletion of sonic hedgehog causes hemiagenesis and ectopic development of the thyroid in mouse. AB - Thyroid dysgenesis encountered in 85% of patients with congenital hypothyroidism is a morphologically heterogeneous condition with primarily unknown pathogenesis. Here we identify sonic hedgehog (Shh) as a novel regulator of thyroid development. In Shh knockout mice the thyroid primordium is correctly specified in the pharyngeal endoderm, but budding and dislocation are slightly delayed. In late development the thyroid fails to form a bilobed gland. Instead a single thyroid mass is found unilaterally and mostly to the left of the midline. Thyroid specific transcription factors (TTF-1 and TTF-2) and thyroglobulin are expressed indicating terminal differentiation. Strikingly, TTF-1- and TTF-2-positive cells aberrantly develop in the presumptive trachea of Shh-/- embryos. The ectopic tissue buds ventrolaterally into the adjacent mesenchyme, and less extensively into the tracheal lumen, forming follicle-like structures that accumulate thyroglobulin. Shh mRNA is not expressed in the thyroid precursor cells at any developmental stage. The results indicate that Shh signaling indirectly governs the symmetric bilobation of the thyroid during late organogenesis. Shh also seems to repress inappropriate thyroid differentiation in nonthyroid embryonic tissues. This study provides clues to the molecular mechanisms that might be dysregulated in thyroid hemiagenesis and development of ectopic thyroid tissue outside the thyroglossal duct. PMID- 15111334 TI - Cultural competence: why? PMID- 15111335 TI - Bisphosphonates and reduction of skeletal events in patients with bone metastatic breast cancer. PMID- 15111336 TI - Challenging paradigms in lymphoma treatment. PMID- 15111337 TI - Fatigue in cancer patients receiving chemotherapy: an analysis of published studies. AB - Fatigue is a subjective experience that affects everybody. In healthy individuals, it can be considered a physiological response to physical or psychological stress. In people with specific diseases, however, fatigue often represents one of the most significant problems. Fatigue can be caused by many factors, both intrinsic to the patient and extrinsic, such as therapeutic interventions. This review, based on published studies, has been conducted with the aim of presenting a critical discussion of the available information on the characteristics, causes and potential treatments of fatigue in cancer patients receiving chemotherapy. The incidence of fatigue in these patients, the methods for measuring and evaluating fatigue, and possible therapeutic options are discussed. An appraisal of the toxicity of various chemotherapeutic agents is also presented. Although fatigue is now an ever more considered aspect of the toxicity of chemotherapy, it remains difficult to establish what standard should be used to make a quali-quantitative evaluation of this symptom. Furthermore, in the absence of a clear demonstration of the efficacy of some therapies, the management of cancer-related fatigue remains poorly defined (except for the treatment of anemia-related fatigue). New randomized clinical trials are necessary to indicate the best strategies for tackling this important problem. PMID- 15111338 TI - Improving information to Italian cancer patients: results of a randomized study. AB - BACKGROUND: It has been widely shown that the provision of adequate levels of information to patients does have a positive effect on quality of life by reducing anxiety and depression levels. The aim of this study was to show how Italian cancer patients rate the information they are given and whether the use of booklets and videotapes can improve their quality of life. PATIENTS AND METHODS: Cancer patients aged between 18 and 80 years who were about to receive their first chemotherapy course were randomized to fill in questionnaires on perceived quality of information, level of psychological distress, perceived severity and curability of the disease, and quality of life. The results were evaluated by means of statistical analyses. RESULTS: Out of 328 consecutive patients enrolled in 21 cancer centers, 86-93% considered the booklets either "very useful" or "useful". The videotape was regarded as "quite" or "much" more complete than the booklets (87%). According to 81%/87% of patients, the information that had been given had improved their knowledge of the disease/chemotherapy either "a lot" or "enough". CONCLUSIONS: The information patients receive from the oncologist was rated the highest, as long as they were devoted enough time. Booklets and videotapes can partially overcome the lack of oral information given by medical doctors. A better informed patient does help the oncologist save time. PMID- 15111339 TI - An oncologist-based model of cancer genetic counselling for hereditary breast and ovarian cancer. AB - BACKGROUND: We describe a multistep model of cancer genetic counselling designed to promote awareness, and disease surveillance and preventive measures for hereditary and familial breast and ovarian cancer. PATIENTS AND METHODS: Step T0 of the model entails information giving; this is followed by pedigree analysis and risk estimation (T1), risk communication and genetic testing (T2), and genetic test result communication (T3). User consent was required to proceed from one step to the next. Surveillance and preventive measures are proposed to at risk users. Of the 311 subjects who requested cancer genetic counselling, consent data to each counselling step were available for 295: 93 were disease-free, 187 had breast cancer, 12 had ovarian cancer and three had breast plus ovarian cancer. RESULTS: Consent was high at T0 (98.39%), T1 (96.40%) and T2 (99.65%). Consent decreased at the crucial points of counselling: T2 (87.71%) and T3 [genetic test result communication (85.08%), and extension of counselling to and testing of relatives (65.36%)]. CONCLUSIONS: The model fosters the user's knowledge about cancer and favours identification of at-risk subjects. Furthermore, by promoting awareness about genetic testing and surveillance measures, the algorithm enables users to make a fully informed choice of action in case of predisposing or familial cancer risk. PMID- 15111340 TI - Assessing websites on complementary and alternative medicine for cancer. AB - BACKGROUND: Many cancer patients search the World Wide Web for information on complementary and alternative medicine. The aim of this study was to evaluate the quality of such information and identify the treatments most frequently discussed. Materials and methods We used a pre-tested search strategy to identify the websites that are most likely to be used by cancer patients and assessed them according to pre-defined criteria. RESULTS: Thirty-two websites were included in this analysis. The quality of these sites was scored on a point system and varied between 8 and 14 points with a maximum of 14. Most sites issued recommendations for a plethora of treatments, which are typically not supported by sound scientific evidence. Three sites had the potential for harming patients through the advice issued. The most frequently discussed therapies were herbal medicines, diets and mind-body therapies. CONCLUSION: The most popular websites on complementary and alternative medicine for cancer offer information of extremely variable quality. Many endorse unproven therapies and some are outright dangerous. PMID- 15111341 TI - Oral ibandronate for the treatment of metastatic bone disease in breast cancer: efficacy and safety results from a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: We report the first results of a randomized trial assessing a new oral aminobisphosphonate, ibandronate, in patients with bone metastases from breast cancer. PATIENTS AND METHODS: Patients (n = 435) received placebo, or oral ibandronate 20 mg or 50 mg once-daily for 96 weeks. The primary efficacy measure was the number of 12-week periods with new bone complications [skeletal morbidity period rate (SMPR)]. Multivariate Poisson regression analysis assessed the relative risk reduction of skeletal-related events. Secondary efficacy analyses included bone pain and analgesic use. Adverse events were monitored. RESULTS: SMPR was significantly reduced with oral ibandronate [placebo 1.2, 20 mg group 0.97 (P = 0.024), 50 mg group 0.98 (P = 0.037)]. Ibandronate 50 mg significantly reduced the need for radiotherapy (P = 0.005 versus placebo). The relative risk of skeletal events was reduced by 38% (20 mg dose) and 39% (50 mg dose) versus placebo (P = 0.009 and P = 0.005). The tolerability profile of ibandronate was similar to placebo. CONCLUSIONS: Oral ibandronate is an effective and well tolerated treatment for metastatic bone disease. The 50 mg dose is being further evaluated in clinical trials, and this dose was recently approved in the European Union for the prevention of skeletal events in patients with breast cancer and bone metastases. PMID- 15111342 TI - A novel continuous infusional 5-fluorouracil-based chemotherapy regimen compared with conventional chemotherapy in the neo-adjuvant treatment of early breast cancer: 5 year results of the TOPIC trial. AB - BACKGROUND: To compare the efficacy of continuous infusional 5-fluorouracil (5 FU)-based chemotherapy against conventional bolus chemotherapy in the preoperative treatment of patients with large operable early breast cancer. PATIENTS AND METHODS: Four hundred and twenty-six women with histologically proven 3 cm invasive early breast cancer were randomised to receive pre operative infusional 5-FU 200 mg/m(2) by daily 24 h continuous infusion via a Hickman line for 18 weeks with epirubicin 60 mg/m(2) intravenous (i.v.) bolus on day 1 and cisplatin 60 mg/m(2) i.v. bolus on day 1, both repeating 3-weekly (infusional ECisF), or conventional bolus doxorubicin 60 mg/m(2) i.v. on day 1 and cyclophosphamide 600 mg/m(2) i.v. on day 1, both repeating 3-weekly (AC), both schedules for six courses. Patients subsequently had local therapy (surgery or radiotherapy or both) and tamoxifen 20 mg orally daily as appropriate. RESULTS: The 5 year results for AC and infusional ECisF, respectively, were as follows: overall response, 75% and 77%; complete clinical remission, 31% and 34%; pathological complete remission (pathCR), 16% for both; and pathCR with residual ductal carcinoma in situ (DCIS), 25% and 24%. Mastectomy rates were 37% and 34%, respectively. Five-year overall survival was 74% for AC and 82% for infusional ECisF (hazard ratio 0.76, 95% confidence interval 0.51-1.13; P = 0.18). Both treatments were well tolerated. Grade III/IV lethargy, vomiting, alopecia and plantar-palmar erythema were significantly greater for infusional ECisF; grade III/IV leucopenia was significantly greater for AC. CONCLUSIONS: Preoperative continuous infusional 5-FU-based chemotherapy is no more active than conventional AC for early breast cancer; with a median 5 year follow-up, the infusion-based schedule shows a non-significant trend towards improved survival. PMID- 15111343 TI - 5-Fluorouracil as protracted continuous intravenous infusion can be added to full dose docetaxel (Taxotere)-cisplatin in advanced gastric carcinoma: a phase I-II trial. AB - BACKGROUND: A phase I-II multicenter trial was conducted to define the maximum tolerated dose (MTD) according to tolerance and toxicity (primary objective), as well as to describe the clinical activity, in terms of response and survival (secondary objectives), of a combination of 5-fluorouracil (5-FU) in protracted continuous intravenous infusion (p.i.v.) with docetaxel and cisplatin for patients with advanced gastric cancer. PATIENTS AND METHODS: Patients with measurable unresectable and/or metastatic gastric carcinoma, World Health Organization performance status < or =1, normal hematological and renal functions, adequate hepatic function and not pretreated for advanced disease by chemotherapy, received up to eight cycles of a combination of docetaxel on day 1, cisplatin on day 1 and 5-FU p.i.v. on days 1-14 (TCF) every 3 weeks, which was escalated up to the MTD, defined by the occurrence of dose-limiting toxicity in two patients in one dose level. RESULTS: Fifty-two patients were accrued and treated (43 in the phase I part of the trial and nine additional at the recommended dose level). A median of five cycles/patient was given. The recommended dose of TCF was docetaxel 85 mg/m(2) on day 1, cisplatin 75 mg/m(2) on day 1 and 5-FU p.i.v. 300 mg/m(2)/day on days 1-14. Grade > or =3 toxicities were neutropenia 79%, alopecia 46%, fatigue 23%, mucositis 10%, diarrhea 19%, nausea/vomiting 13%, neurological 4% and palmar-plantar 2%. Ten non-fatal febrile neutropenia episodes were recorded in eight patients. There were no treatment related deaths. Among 41 patients with measurable disease (79%), we observed one complete and 20 partial responses for an overall intent-to-treat response rate of 51% (95% confidence interval 35-67%). Five patients (20%) had stable disease for > or =12 weeks (four cycles). The median overall survival was 9.3 months. CONCLUSIONS: 5-FU p.i.v. at 300 mg/m(2)/day for 2 weeks out of three could be safely added to the docetaxel-cisplatin (TC) combination, but the dose of docetaxel had to be reduced to 75 mg/m(2) in a subsequent phase II trial. This drug regimen seems to be very active in advanced gastric cancer. Comparison with both TC and ECF in a randomized SAKK trial is ongoing. PMID- 15111344 TI - Combination of folinic acid, 5-fluorouracil bolus and infusion, and cisplatin (LV5FU2-P regimen) in patients with advanced gastric or gastroesophageal junction carcinoma. AB - BACKGROUND: Combination chemotherapy with continuous 5-fluorouracil (5-FU) and cisplatin in a monthly regimen is one of the standard treatments for advanced gastric carcinoma. This study evaluated the new LV5FU2-P regimen, designed to improve efficacy and tolerance of the 5-FU plus cisplatin combination. PATIENTS AND METHODS: Forty-three patients with advanced or metastatic gastroesophageal junction or gastric carcinoma were prospectively included in the study. They were treated every 14 days with cisplatin 50 mg/m(2) on day 2 plus folinic acid 200 mg/m(2)/day as a 2-h intravenous (i.v.) infusion on days 1 and 2, plus bolus 5-FU 400 mg/m(2)/day on days 1 and 2, plus continuous 5-FU 600 mg/m(2)/day as a 22-h i.v. infusion on days 1 and 2. Ten patients received a simplified regimen (folinic acid 40 mg/m(2) day 1 + bolus 5-FU 400 mg/m(2) day 1 + continuous 5-FU 2400 mg/m(2) on days 1 and 2 with cisplatin 50 mg/m(2) on day 2). RESULTS: All the patients were assessable for response and 42 for toxicity. One patient achieved a complete response and 15 a partial response, for an overall response rate of 37.2% [95% confidence interval (CI) 22.1% to 52.3%]. The median progression-free survival was 7.2 months (95% CI 5.4-10.9) and the overall survival was 13.3 months (95% CI 10.1-16.4). There were no treatment-related deaths. Hematological and gastrointestinal toxicities were the most common severe toxicities. CONCLUSIONS: LV5FU2-P is an active and well tolerated regimen in the treatment of advanced gastroesophageal junction or gastric carcinomas. It warrants evaluation comparatively with other active regimens. PMID- 15111345 TI - Gemcitabine concurrent with continuous infusional 5-fluorouracil in advanced biliary cancers: a review of the Princess Margaret Hospital experience. AB - BACKGROUND: Unresectable biliary tract cancer has a very poor prognosis. A combination of weekly gemcitabine plus continuous infusional 5-fluorouracil (5 FU) (GEM/CVI 5-FU) was evaluated as therapy for this cancer. PATIENTS AND METHODS: The charts of 27 patients with advanced biliary tract adenocarcinoma treated with GEM/CVI 5-FU at the Princess Margaret Hospital were evaluated for response, survival and toxicity. The treatment consisted of a 30-min infusion of gemcitabine at 900 mg/m(2) on days 1, 8 and 15 of a 28-day cycle plus 5-FU given via a peripherally inserted central line at 200 mg/m(2)/day continuously for 21 days, every 28 days. RESULTS: Objective responses were observed in nine patients (33%; 95% confidence interval 17% to 54%). An additional eight patients (30%) achieved stable disease for a median of 4 months (range 2.3-11). Median time to progression and overall survival were 3.7 and 5.3 months, respectively. Direct chemotherapy-related toxicity was mild, with only 11% grade > or =3 myelosuppression. Central venous catheter complications were common (26%). There were no treatment-related deaths. CONCLUSIONS: This study shows that GEM/CVI 5-FU is active and well tolerated in advanced and metastatic biliary tract cancers. PMID- 15111346 TI - Induction of complete tumor necrosis may reduce intrahepatic metastasis and prolong survival in patients with hepatocellular carcinoma undergoing locoregional therapy: a prospective study. AB - BACKGROUND: Transarterial chemoembolization (TACE) and percutaneous acetic acid injection (PAI) are effective locoregional therapies for hepatocellular carcinoma (HCC). This study aimed to investigate whether HCC patients who had initial complete response to these treatments had a subsequent lower risk of intrahepatic metastasis. PATIENTS AND METHODS: A total of 152 patients who underwent locoregional therapy (94 received PAI and 58 received both TACE and PAI) for HCC (tumor size < or =5 cm) were prospectively evaluated. RESULTS: In all, 60 (39%) patients had a complete tumor necrosis after treatment. The cumulative incidence of the development of intrahepatic metastasis was lower for patients with complete remission (P = 0.005) and for patients with smaller (< or =3 cm) tumor size (P = 0.083). Cox multivariate survival analysis showed that absence of complete remission [relative risk (RR) 2.7; 95% confidence interval (CI) 1.4-5.3; P = 0.003] was the only independent factor that predicted the occurrence of intrahepatic metastasis. Patients with complete remission had a significantly better long-term survival than those without (P = 0.002), and the occurrence of intrahepatic metastasis over time independently predicted a decreased survival (RR 3.2; 95% CI 2.0-6.1; P = 0.019). CONCLUSIONS: Induction of complete tumor necrosis in HCC patients undergoing locoregional therapy may decrease the risk of intrahepatic metastasis and improve survival. PMID- 15111347 TI - Efficacy of intraperitoneal chemohyperthermia with oxaliplatin in colorectal peritoneal carcinomatosis. Preliminary results in 24 patients. AB - BACKGROUND: The complete resection of macroscopic colorectal peritoneal carcinomatosis (PC), followed by intraoperative intraperitoneal chemohyperthermia (IPCH) to treat residual microscopic disease, leads to cure in some patients. We report preliminary results on survival in a phase II study using oxaliplatin (LOHP). PATIENTS AND METHODS: Twenty-four patients with macroscopic colorectal PC underwent complete resection of the PC followed by IPCH with LOHP performed in an open abdominal cavity. The dose of LOHP was 460 mg/m(2) in 2 l/m(2), during 30 min at 43 degrees C, at a flow rate of 2 l/min. During the hour preceding IPCH, they received an intravenous administration of 5-fluorouracil (400 mg/m(2)) and leucovorin (20 mg/m(2)). RESULTS: Mean peritoneal tumoral extension (Sugarbaker's Index) was 16.9 +/- 9.5, median operative duration was 490 min and median blood loss was 965 ml. There were two postoperative deaths (8%) by intracerebral hemorrhage, and morbidity rate was 41.6%. Minimal follow-up was 18 months and median follow-up was 27.4 months (range 18.3-49.6). At 1, 2 and 3 years, overall survival rates were 83%, 74% and 65%, and disease-free survival rates were 70%, 50% and 50%, respectively. Only 32% of the 22 postoperative living patients presented a peritoneal recurrence. A peritoneal index >24 influenced survival, with a 17% recurrence rate at 2 years versus 63% when it was <24 (P = 0.005). CONCLUSION: This new modality of treatment, when feasible, gives encouraging preliminary results, with a promising 3-year survival rate of 65%. PMID- 15111348 TI - Gefitinib as a last treatment option for non-small-cell lung cancer: durable disease control in a subset of patients. AB - BACKGROUND: We describe 16 months' single-institution experience with gefitinib ("Iressa", ZD1839) used as "ultimum refugium" for pretreated non-small-cell lung cancer (NSCLC) patients. PATIENTS AND METHODS: Toxicity, response and survival data of NSCLC patients participating in a compassionate-use program with gefitinib were reviewed. Documented disease progression and confirmation of the absence of other treatment options were requested. Oral gefitinib at a dose of 250 mg/day was given until disease progression, unacceptable toxicity or death. Cox's proportional hazards model was used to analyze relationships between factors and probability of survival. RESULTS: Rapid disease precluded treatment in eight cases. Of 92 evaluable patients, one-third had a baseline performance status (PS) of > or =2. The main side-effects of gefitinib were grade 1-2 diarrhea and skin rash. A disease control rate of 46% (objective response rate 8.7%) and 1-year survival of 29% were documented. Histology (adenocarcinoma) and a "never-smoking" history were predictive of response. Number of previous chemotherapy regimens, gender, time since diagnosis and time since last chemotherapy lacked such an association. Radiotherapy during gefitinib treatment was well tolerated and was associated with prolonged survival in a patient with multiple brain metastases. Multivariate analyses revealed a significant impact of PS on survival. A "never-smoking" history, adenocarcinoma/bronchoalveolar-cell carcinoma and female gender showed a trend towards better survival outcomes. CONCLUSION: Gefitinib's single-agent activity in a group consisting of pretreated NSCLC patients is confirmed. Side-effects of gefitinib were mild. Prolonged survival was associated with good PS and less significantly with a never-smoking history, female gender and histology. Additional studies on mechanisms of tumor control and selection of target populations for this remarkable new drug are warranted. PMID- 15111349 TI - Joinpoint regression analysis of lung cancer mortality, Andalusia 1975-2000. AB - BACKGROUND: Previous analyses of lung cancer mortality were based on models of death rates within one time period, assuming that rates increase or decrease with time at a constant rate. The aim of this work is to analyse recent changes in lung cancer mortality trends in Andalusia (Spain) during the period of 1975-2000 using joinpoint regression models. PATIENTS AND METHODS: Mortality data were obtained from the Death Registry of Andalusia. For each gender, age group specific and standardised (overall and truncated) rates were calculated by the direct method (using the world standard population). The joinpoint analysis was used to identify the best-fitting points where a statistically significant change in the trend occurred. RESULTS: Lung cancer is the leading cause of cancer mortality in men, with an increasing trend up to 1994. After that year, rates began to decrease significantly (-1.8% yearly from 1994 to 2000). Standardised rates in women exhibited a downward trend until the early 1990s, after which they levelled off (overall standardised rates) or increased significantly (truncated rates 35-64 years). CONCLUSIONS: An increase in lung cancer mortality has been observed in young women. There seems to be a relationship with the prevalence in smoking in men and women. PMID- 15111350 TI - Expression of Cox-2 protein in radioresistant laryngeal cancer. AB - BACKGROUND: Radiotherapy is the principal modality used to treat early stage laryngeal cancer. Unfortunately treatment failures occur in 10-25% of patients. Subsequent salvage surgery is technically more difficult, with increased complication and failure rates. The ability to predict or prevent radioresistance would improve the poor survival associated with this disease. Cox-2 is an inducible enzyme involved with prostaglandin synthesis. We investigated a potential role for Cox-2 in predicting radioresistance in laryngeal cancer. PATIENTS AND METHODS: Using immunohistochemical techniques we examined the expression of Cox-2 protein in 122 pre-treatment laryngeal biopsies. All tumours were treated with single modality radiotherapy (curative intent). The group comprised of 61 radioresistant and 61 radiosensitive tumours matched for T stage, laryngeal subsite, gender and smoking history. RESULTS: Cox-2 expression was detected in 41 of 61 (67%) biopsy samples from patients with radioresistant tumours and 25 of 61 (41%) radiosensitive tumours. Overexpression was significantly associated with radioresistant tumours (P = 0.004). Cox-2 has a 67% accuracy in predicting radiotherapy failure. CONCLUSION: Cox-2 may have prognostic value in predicting response to radiotherapy. Cox-2 inhibitors such as NS-398 have been shown to enhance the effects of radiotherapy. We suggest that their use may be beneficial in patients who are destined to fail radiotherapy. PMID- 15111351 TI - Effect of cervical spinal cord stimulation on regional blood flow and oxygenation in advanced head and neck tumours. AB - BACKGROUND: Tumour ischaemia leads to decreased delivery of oxygen, chemotherapy and radiosensitisers. Hypoxia in head and neck (H&N) tumours is an important adverse prognostic factor. Spinal cord stimulation (SCS) is a well-established neurosurgical technique in the treatment of several ischaemic syndromes. This prospective study evaluated the effect of cervical-SCS on common carotid artery (CCA) blood flow and tumour oxygenation in patients with advanced H&N cancer. PATIENTS AND METHODS: Sixteen patients with advanced H&N tumours were enrolled. Cervical-SCS devices were inserted subcutaneously prior to commencement of scheduled chemoradiotherapy. Pre- and post-SCS measurements were as follows: (i) tumour oxygenation (mmHg) using polarographic probes; (ii) blood flow quantification (ml/min) and diastolic and systolic velocimetry (cm/s) in the CCA using colour Doppler. RESULTS: After SCS, median tumour oxygenation increased in two-thirds of patients (34%; P = 0.023), all patients had improved CCA blood flow (50%; P <0.001) and almost all patients showed an increased CCA diastolic velocity (26%; P = 0.003) and systolic velocity (20%; P = 0.011). CONCLUSIONS: Cervical-SCS increased tumour oxygenation and CCA blood flow, and could enhance the loco-regional delivery of oxygen, radiosensitising and chemotherapeutic drugs. Cervical-SCS as adjuvant in chemoradiotherapy of these tumours warrants further investigation. PMID- 15111352 TI - Survival in nephroblastoma treated according to the trial and study SIOP-9/GPOH with respect to relapse and morbidity. AB - BACKGROUND: Recent Wilms' tumor (WT) trials and studies have tried to determine the minimal therapy needed for cure. The goal was survival without morbidity. PATIENTS AND METHODS: From January 1989 to March 1994 the German Society of Pediatric Oncology and Hematology registered 440 patients (median age 2.9 years; 231 male, 209 female) with WTs (preoperative chemotherapy 362) for therapy according to the International Society of Pediatric Oncology Trial and Study 9. Therapy for relapse depended on site of relapse and therapy already received. Follow-up included inquiries for morbidity. Prognostic factors for relapse and death were evaluated. RESULTS: Five-year survival of WTs was 89.5%; 98.2% (385 of 392) of survivors had a follow-up of 5 years (range 0.8-12.6; median 8). In non anaplastic WTs, young age (<2 years) was of significance (P = 0.026) for a better survival. Non-anaplastic WTs (407 patients) had a 5-year survival of 92.3%, versus 48.5% in anaplastic WTs (33 patients), and a 5-year relapse-free survival of 87.6% versus 42.4%. Survival after relapse was significantly worse for anaplastic than for non-anaplastic WTs (residual 3-year survival 11.8% versus 54.3%; P <0.0001). In preoperatively treated WTs, anaplasia was a strong prognostic factor for death [relative risk (RR) 4.7], followed by poor response to preoperative therapy (RR 3.6), stage IV (RR 3.2) and abdominal stage III (RR 2.2). Low abdominal stages (12. Later in the winter, however, animals exposed to both photoperiodic treatments showed strong rhythms of foraging activity irrespective of the imposed photoperiod. It is suggested that the autumn cessation may maximize the fitness of N. virens, a spring-breeding semelparous organism, by reducing risk during gamete maturation, while spontaneous resurgence of activity after the winter solstice permits animals that are not physiologically competent to spawn to accrue further metabolic reserves. This response is believed to be initiated by a seasonal (possibly circannual) endogenous oscillator or interval timer. PMID- 15111366 TI - Adaptable defense: a nudibranch mucus inhibits nematocyst discharge and changes with prey type. AB - Nudibranchs that feed on cnidarians must defend themselves from the prey's nematocysts or risk their own injury or death. While a nudibranch's mucus has been thought to protect the animal from nematocyst discharge, an inhibition of discharge by nudibranch mucus has never been shown. The current study investigated whether mucus from the aeolid nudibranch Aeolidia papillosa would inhibit nematocyst discharge from four species of sea anemone prey. Sea anemone tentacles were contacted with mucus-coated gelatin probes, and nematocyst discharge was quantified and compared with control probes of gelatin only. Mucus from A. papillosa inhibited the discharge of nematocysts from sea anemone tentacles. This inhibition was specifically limited to the anemone species on which the nudibranch had been feeding. When the prey species was changed, the mucus changed within 2 weeks to inhibit the nematocyst discharge of the new prey species. The nudibranchs apparently produce the inhibitory mucus rather than simply becoming coated in anemone mucus during feeding. Because of the intimate association between most aeolid nudibranchs and their prey, an adaptable mucus protection could have a significant impact on the behavior, distribution, and life history of the nudibranchs. PMID- 15111367 TI - Humor in the physician-patient encounter. PMID- 15111368 TI - Death of an Arabian Jew. AB - A 69-year-old Judean man presents with chronic low-grade fever, pedal edema, and abdominal pain. His condition deteriorates over several weeks with the appearance of shortness and foulness of breath, pruritus, convulsions of every limb, and gangrene of the genitalia. Just before he dies, he orders dozens of the leading men of his kingdom imprisoned and instructs his sister to kill them all after he is gone. Who is he and what is the likely cause of his death? PMID- 15111370 TI - Use of aspirin and ibuprofen compared with aspirin alone and the risk of myocardial infarction. AB - BACKGROUND: Laboratory investigations suggest that the simultaneous use of aspirin and ibuprofen may attenuate the antiplatelet effect of aspirin, making it less useful for cardioprotection. To determine if there is clinical evidence of this potentially harmful interaction, we conducted a retrospective matched case control study. METHODS: All patients issued outpatient prescriptions for aspirin or ibuprofen from January 1, 1990, to December 31, 2000, at the Durham Veterans Affairs Medical Center pharmacy were included in the study. Patients who used aspirin and ibuprofen concurrently were matched against those who used aspirin only by race, sex, age within 10 years, and cholesterol levels (either low density lipoprotein or total cholesterol) to within 30 mg/dL (0.78 mmol/L). The rate ratio of experiencing a myocardial infarction per patient-month of drug exposure was then determined. RESULTS: Some 3859 patients received both aspirin and ibuprofen, for a total of 52 139 patient-months of medication use. This group experienced 138 infarctions. The 10 239 patients receiving aspirin only, for a total of 156 417 patient-months of use, experienced 684 infarctions. The rate ratio of having an infarction was 0.61 (95% confidence interval, 0.50-0.73) (P <.001), favoring the group that took aspirin and ibuprofen simultaneously. An analysis of diabetic patients found a rate ratio of 0.48 (95% confidence interval, 0.34-0.66) (P <.001). An examination of patients who spent time in both groups at different times resulted in a rate ratio of infarction during combined use of 0.70 (95% confidence interval, 0.59-0.83) (P <.001). CONCLUSION: There does not seem to be an increased risk of myocardial infarction among patients simultaneously consuming aspirin and ibuprofen compared with aspirin alone. PMID- 15111369 TI - Catheter-associated infections: pathogenesis affects prevention. AB - Intravascular catheters and urinary catheters are the 2 most commonly inserted medical devices in the United States, and they are likewise the two most common causes of nosocomially acquired bloodstream infection. Biofilm formation on the surfaces of indwelling catheters is central to the pathogenesis of infection of both types of catheters. The cornerstone to any preventive strategy of intravascular catheter infections is strict attention to infection control practices. Antimicrobial-impregnated intravascular catheters are a useful adjunction to infection control measures. Prevention of urinary catheter associated infection is hindered by the numbers and types of organisms present in the periurethral area as well as by the typically longer duration of catheter placement. Antimicrobial agents in general have not been effective in preventing catheter-associated urinary tract infection in persons with long-term, indwelling urethral catheters. Preventive strategies that avoid the use of antimicrobial agents may be necessary in this population. PMID- 15111371 TI - Prayer for health concerns: results of a national survey on prevalence and patterns of use. AB - BACKGROUND: Prayer is a common practice in the United States, yet little is known about the prevalence and patterns of use of prayer for health concerns. OBJECTIVE: To determine the prevalence and patterns of use of prayer for health concerns. METHODS: We conducted a national survey in 1998 (N = 2055, 60% weighted response rate) on use of prayer. Data were also collected on sociodemographics, use of conventional medicine, and use of complementary and alternative medical therapies. Factors associated with the use of prayer were analyzed using multivariable logistic regression. RESULTS: We found that 35% of respondents used prayer for health concerns; 75% of these prayed for wellness, and 22% prayed for specific medical conditions. Of those praying for specific medical conditions, 69% found prayer very helpful. Factors independently associated with increased use of prayer (P<.05) included age older than 33 years (age 34-53 years: odds ratio [OR], 1.6 [95% confidence interval (CI), 1.3-2.1]; age > or =54 years: OR, 1.5 [95% CI, 1.1-2.0]); female sex (OR, 1.4 [95% CI, 1.1-1.7]); education beyond high school (OR, 1.5 [95% CI, 1.2-1.8]); and having depression, chronic headaches, back and/or neck pain, digestive problems, or allergies. Only 11% of respondents using prayer discussed it with their physicians. CONCLUSIONS: An estimated one third of adults used prayer for health concerns in 1998. Most respondents did not discuss prayer with their physicians. Prayer was used frequently for common medical conditions, and users reported high levels of perceived helpfulness. PMID- 15111372 TI - Seasonal variation in serum cholesterol levels: treatment implications and possible mechanisms. AB - BACKGROUND: A variety of studies have noted seasonal variation in blood lipid levels. Although the mechanism for this phenomenon is not clear, such variation could result in larger numbers of people being diagnosed as having hypercholesterolemia during the winter. METHODS: We conducted a longitudinal study of seasonal variation in lipid levels in 517 healthy volunteers from a health maintenance organization serving central Massachusetts. Data collected during a 12-month period for each individual included baseline demographics and quarterly anthropometric, blood lipid, dietary, physical activity, light exposure, and behavioral information. Data were analyzed using sinusoidal regression modeling techniques. RESULTS: The average total cholesterol level was 222 mg/dL (5.75 mmol/L) in men and 213 mg/dL (5.52 mmol/L) in women. Amplitude of seasonal variation was 3.9 mg/dL (0.10 mmol/L) in men, with a peak in December, and 5.4 mg/dL (0.14 mmol/L) in women, with a peak in January. Seasonal amplitude was greater in hypercholesterolemic participants. Seasonal changes in plasma volume explained a substantial proportion of the observed variation. Overall, 22% more participants had total cholesterol levels of 240 mg/dL or greater (> or =6.22 mmol/L) in the winter than in the summer. CONCLUSIONS: This study confirms seasonal variation in blood lipid levels and suggests greater amplitude in seasonal variability in women and hypercholesterolemic individuals, with changes in plasma volume accounting for much of the variation. A relative plasma hypervolemia during the summer seems to be linked to increases in temperature and/or physical activity. These findings have implications for lipid screening guidelines. Further research is needed to better understand the effects of a relative winter hemoconcentration. PMID- 15111373 TI - A comparison of the effects of raloxifene and conjugated equine estrogen on bone and lipids in healthy postmenopausal women. AB - BACKGROUND: Although many studies have assessed the effects of estrogen and raloxifene hydrochloride on bone mineral density and serum lipid concentrations, there are few direct comparative data. METHODS: Randomized placebo-controlled trial for 3 years, intention-to-treat analysis. Six hundred nineteen postmenopausal women with prior hysterectomy (mean age, 53.0 years) were studied in 38 centers in Europe, North America, Australasia, and South Africa. They were randomized to 60 mg/d or 150 mg/d of raloxifene, 0.625 mg/d of conjugated equine estrogen (CEE), or placebo. Bone density of the lumbar spine and proximal femur, biochemical markers of bone turnover, and fasting serum lipid concentrations were assessed for 3 years. RESULTS: Compared with baseline, bone density in the lumbar spine progressively declined by 2.0% in the placebo group (P <.05), was stable in the 2 raloxifene groups, and increased 4.6% in the subjects receiving CEE (P <.001). Effects in both raloxifene groups were different from those observed in the CEE and placebo groups (P <.001). Bone density in the total hip showed similar results. Conjugated equine estrogen produced significantly greater depression of serum osteocalcin, bone-specific alkaline phosphatase, and urine C telopeptide, compared with raloxifene. Each of the active treatments caused comparable depression of low-density lipoprotein cholesterol below placebo levels (P <.001 at most time points). Raloxifene did not affect high-density lipoprotein cholesterol, whereas CEE increased it by 13.4% compared with placebo at 3 years (P <.001). Triglyceride concentrations increased 24.6% in the CEE group at 3 years (P <.003), a significantly greater change than in the raloxifene groups, which were 4.9% and 8.0% above baseline (P < or =.002) but not different from placebo. Urinary incontinence was reported in 11 women receiving CEE, but in only 1 or 2 in each of the other groups (P < or =.01 compared with the other groups). Hernias occurred less frequently in those receiving 150 mg/d of raloxifene or CEE (P =.03 vs placebo). CONCLUSIONS: Raloxifene and CEE have beneficial effects on bone density and bone turnover, although effects of CEE are more marked. Raloxifene and CEE produce different patterns of lipid responses and have distinct adverse effect profiles. PMID- 15111374 TI - The effect of warfarin and intensity of anticoagulation on outcome of intracerebral hemorrhage. AB - BACKGROUND: Warfarin sodium is highly effective for prevention of embolic stroke, particularly in nonvalvular atrial fibrillation, but its expected benefit can be offset by risk of intracerebral hemorrhage (ICH). We studied the determinants of ICH outcome to quantify the independent effect of warfarin. METHODS: Consecutive patients with supratentorial ICH treated in a tertiary care hospital with a neurointensive care unit were prospectively identified during a 7-year period, and data on hemorrhage location, clinical characteristics, and warfarin use were collected. Independent predictors of 3-month mortality were determined using multiple logistic regression analysis. RESULTS: Of 435 consecutive patients aged 55 years or older, 102 (23.4%) were taking warfarin at the time of ICH. Three month mortality was 25.8% for those not taking warfarin and 52.0% for those taking warfarin. Independent predictors of death were warfarin use (odds ratio [OR], 2.2; 95% confidence interval [CI], 1.3-3.8), age 70 years or older (OR, 2.4; 95% CI, 1.4-4.0), and presence of diabetes mellitus (OR, 1.8; 95% CI, 1.0 3.3). Although 68.0% of all warfarin-related hemorrhages occurred at an international normalized ratio (INR) of 3.0 or less, increasing degrees of anticoagulation were strongly associated with increasing risk of death compared with no warfarin use. CONCLUSIONS: Patients taking warfarin had a doubling in the rate of intracerebral hemorrhage mortality in a dose-dependent manner. The data suggest that careful control of the INR, already known to limit the risk of warfarin-related ICH, may also limit its severity. PMID- 15111375 TI - Dietary factors and the risk of incident kidney stones in younger women: Nurses' Health Study II. AB - BACKGROUND: In older women and men, greater intakes of dietary calcium, potassium, and total fluid reduce the risk of kidney stone formation, while supplemental calcium, sodium, animal protein, and sucrose may increase the risk. Recently, phytate has been suggested to play a role in stone formation. To our knowledge, no prospective information on the role of dietary factors and risk of kidney stone formation is available in younger women. METHODS: We prospectively examined, during an 8-year period, the association between dietary factors and the risk of incident symptomatic kidney stones among 96 245 female participants in the Nurses' Health Study II; the participants were aged 27 to 44 years and had no history of kidney stones. Self-administered food frequency questionnaires were used to assess diet in 1991 and 1995. The main outcome measure was an incident symptomatic kidney stone. Cox proportional hazards regression models were used to adjust simultaneously for various risk factors. RESULTS: We documented 1223 incident symptomatic kidney stones during 685 973 person-years of follow-up. After adjusting for relevant risk factors, a higher dietary calcium intake was associated with a reduced risk of kidney stones (P =.007 for trend). The multivariate relative risk among women in the highest quintile of intake of dietary calcium compared with women in the lowest quintile was 0.73 (95% confidence interval, 0.59-0.90). Supplemental calcium intake was not associated with risk of stone formation. Phytate intake was associated with a reduced risk of stone formation. Compared with women in the lowest quintile of phytate intake, the relative risk for those in the highest quintile was 0.63 (95% confidence interval, 0.51-0.78). Other dietary factors showed the following relative risks (95% confidence intervals) among women in the highest quintile of intake compared with those in the lowest quintile: animal protein, 0.84 (0.68-1.04); fluid, 0.68 (0.56-0.83); and sucrose, 1.31 (1.07-1.60). The intakes of sodium, potassium, and magnesium were not independently associated with risk after adjusting for other dietary factors. CONCLUSIONS: A higher intake of dietary calcium decreases the risk of kidney stone formation in younger women, but supplemental calcium is not associated with risk. This study also suggests that some dietary risk factors may differ by age and sex. Finally, dietary phytate may be a new, important, and safe addition to our options for stone prevention. PMID- 15111376 TI - Physical activity, body mass index, and risk of type 2 diabetes in patients with normal or impaired glucose regulation. AB - BACKGROUND: Sedentary lifestyle, obesity, and impaired glucose regulation are associated with the risk of type 2 diabetes. However, the joint associations of these risk factors are not known. METHODS: We prospectively followed up 2017 Finnish men and 2352 Finnish women aged between 45 and 64 years without a history of known or newly diagnosed diabetes at baseline. Single and joint associations of physical activity, body mass index (BMI), and blood glucose levels with risk of type 2 diabetes were examined using Cox proportional hazards models. RESULTS: During a mean follow-up of 9.4 years, there were 120 incident cases of type 2 diabetes. After adjustment for confounding factors (age, study year, sex, systolic blood pressure, smoking, and education), physical activity was found to be inversely associated with the risk of type 2 diabetes. This association was persistent in subjects with (1) both obesity and impaired glucose regulation, (2) either obesity or impaired glucose regulation, and (3) a normal BMI and glucose regulation. Similarly, the multivariate-adjusted positive association between BMI and risk of type 2 diabetes was consistently observed. Obesity in subjects who reported being inactive and had normal glucose levels was associated with an increased risk of diabetes compared with a normal BMI in subjects who reported being active and had impaired glucose regulation. CONCLUSIONS: Increasing physical activity can reduce the risk of type 2 diabetes. The protective effect of physical activity was observed in subjects with an excessive BMI and elevated glucose levels. Physical activity and weight control are critical factors in diabetes prevention in subjects with both normal and impaired blood glucose regulation. PMID- 15111377 TI - Effects of testosterone administration in human immunodeficiency virus-infected women with low weight: a randomized placebo-controlled study. AB - BACKGROUND: The prevalence of human immunodeficiency virus (HIV) disease is increasing among women, many of whom remain symptomatic with low weight and poor functional status. Although androgen levels may often be reduced in such patients, the safety, tolerability, and efficacy of testosterone administration in this population remains unknown. METHODS: A total of 57 HIV-infected women with free testosterone levels less than the median of the reference range and weight less than 90% of ideal body weight or weight loss greater than 10% were randomly assigned to receive transdermal testosterone (4 mg/patch) twice weekly or placebo for 6 months. Muscle mass was assessed by urinary creatinine excretion. Muscle function was assessed by the Tufts Quantitative Muscle Function Test. Treatment effect at 6 months was determined by analysis of covariance. Results are mean +/- SEM unless otherwise specified. RESULTS: At baseline, subjects were low weight (body mass index [calculated as weight in kilograms divided by the square of height in meters] 20.6 +/- 0.4), with significant weight loss from pre-illness maximum weight (18.7% +/- 1.2%), and demonstrated reduced muscle function (upper and lower extremity muscle strength, 83% and 67%, respectively, of predicted range). Testosterone treatment resulted in significant increases in testosterone levels vs placebo (total testosterone: 37 +/- 5 vs -2 +/- 2 ng/dL [1.3 +/- 0.2 vs -0.1 +/- 0.1 nmol/L] [P<.001]; free testosterone: 3.7 +/- 0.5 vs -0.4 +/- 0.3 pg/mL [12.8 +/- 1.7 vs -1.4 vs 1.0 pmol/L] [P<.001]) and was well tolerated, without adverse effects on immune function, lipid and glucose levels, liver function, or body composition or the adverse effect of hirsutism. Muscle mass tended to increase (1.4 +/- 0.6 vs 0.3 +/- 0.8 kg; P =.08), and shoulder flexion (0.4 +/- 0.3 vs -0.5 +/- 0.3 kg; P =.02), elbow flexion (0.3 +/- 0.4 vs -0.7 +/- 0.4 kg; P =.04), knee extension (0.2 +/- 1.0 vs -1.7 +/- 1.3 kg; P =.02), and knee flexion (0.7 +/- 0.5 vs 0.3 +/- 0.7 kg; P =.04) increased in the testosterone-treated compared with the placebo-treated subjects. CONCLUSIONS: Testosterone administration is well-tolerated and increases muscle strength in low-weight HIV-infected women. Testosterone administration may be a useful adjunctive therapy to maintain muscle function in symptomatic HIV-infected women. PMID- 15111378 TI - Renal tubular acidosis, Sjogren syndrome, and bone disease. AB - BACKGROUND: There has been disagreement about whether osteomalacia (adult rickets) occurs in adults with type 1 (distal) renal tubular acidosis (RTA1). Therefore, after finding scapular pseudofractures in a patient with RTA1 and Sjogren syndrome, we decided to survey other patients with RTA to learn whether osteomalacia occurred in others and, if it did, whether it was necessarily associated with the presence of Sjogren syndrome. METHODS: We examined the hospital records and laboratory findings of 250 patients with codes for RTA, 124 with codes for osteomalacia, and 20 with codes for Sjogren syndrome who were seen at a university-affiliated acute care municipal hospital since 1990. Further detailed survey was then limited to patients older than 15 years and excluded those with potentially confounding causes of bone disease such as chronic renal insufficiency or sickle cell disease. Seven adults with RTA1 were thereby identified. RESULTS: Two adults with RTA1 had radiological and biochemical findings compatible with osteomalacia, and 1 had findings compatible with Sjogren syndrome. A third patient without Sjogren syndrome had biochemical findings suggestive of osteomalacia. CONCLUSIONS: Osteomalacia seems to occur in some adult patients with RTA1, and not only in association with Sjogren syndrome. We found no biochemical evidence of osteomalacia in the patients with Sjogren syndrome who did not have RTA. PMID- 15111379 TI - Angioedema associated with angiotensin-converting enzyme inhibitor use: outcome after switching to a different treatment. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) inhibitors are associated with angioedema episodes that are potentially life-threatening. Few data are available on the outcome of patients reporting this adverse effect when they are switched to another drug. Scattered reports of angioedema associated with angiotensin II receptor blocker (ARB) use question the safety of using these drugs in patients with ACE inhibitor-related angioedema. We describe 64 consecutive patients with ACE inhibitor-related angioedema, the outcome after discontinuing this treatment, and the safety of using ARBs. METHODS: Retrospective analysis of 64 consecutive patients (January 1993 to June 2002) presenting with angioedema onset while receiving treatment with an ACE inhibitor. RESULTS: Patients were recommended to stop ACE inhibitor use, substituting it upon advice of the physician. Fifty-four patients were available for follow-up (median follow-up, 11 months; range, 1-80 months): 26 had switched to an ARB, 14 to a calcium antagonist, and 14 to other antihypertensive drugs. Angioedema disappeared or drastically reduced upon withdrawal of the ACE inhibitor in 46 patients (85%). For the remaining 8 patients, angioedema was due to a cause other than ACE inhibitor use in 2; angioedema persisted independent of the treatment and without apparent cause (idiopathic angioedema) in 4; angioedema persisted after switching to an ARB and disappeared upon its withdrawal in 2. CONCLUSIONS: Stopping ACE inhibitor use without further assessments is a successful measure in the large majority of patients developing angioedema while taking this drug. Only a small percentage of patients with ACE inhibitor-related angioedema continue with this symptom when switched to an ARB. PMID- 15111380 TI - The "improbability" of complementary and alternative medicine. PMID- 15111381 TI - Religious conviction and decisions near the end of life. PMID- 15111382 TI - The patient's right to faith. PMID- 15111383 TI - Muslim and Jewish perspectives on inappropriate treatment at the end of life. PMID- 15111384 TI - Melting under stress. PMID- 15111385 TI - Pulling-speed-dependent force-extension profiles for semiflexible chains. AB - We present theory and simulations to describe nonequilibrium stretching of semiflexible chains that serve as models of DNA molecules. Using a self consistent dynamical variational approach, we calculate the force-extension curves for worm-like chains as a function of the pulling speed, v(0). Due to nonequilibrium effects the stretching force, which increases with v(0), shows nonmonotonic variations as the persistence length increases. To complement the theoretical calculations we also present Langevin simulation results for extensible worm-like chain models for the dynamics of stretching. The theoretical force-extension predictions compare well with the simulation results. The simulations show that, at high enough pulling speeds, the propagation of tension along the chain conformations transverse to the applied force occurs by the Brochard-Wyart's stem-flower mechanism. The predicted nonequilibrium effects can only be observed in double-stranded DNA at large ( approximately 100 microm/s) pulling speeds. PMID- 15111386 TI - Dynamic receptor team formation can explain the high signal transduction gain in Escherichia coli. AB - Evolution has provided many organisms with sophisticated sensory systems that enable them to respond to signals in their environment. The response frequently involves alteration in the pattern of movement, either by directed movement, a process called taxis, or by altering the speed or frequency of turning, which is called kinesis. Chemokinesis has been most thoroughly studied in the peritrichous bacterium Escherichia coli, which has four helical flagella distributed over the cell surface, and swims by rotating them. When rotated counterclockwise the flagella coalesce into a propulsive bundle, producing a relatively straight "run," and when rotated clockwise they fly apart, resulting in a "tumble" which reorients the cell with little translocation. A stochastic process generates the runs and tumbles, and in a chemoeffector gradient, runs that carry the cell in a favorable direction are extended. The cell senses spatial gradients as temporal changes in receptor occupancy and changes the probability of counterclockwise rotation (the bias) on a fast timescale, but adaptation returns the bias to baseline on a slow timescale, enabling the cell to detect and respond to further concentration changes. The overall structure of the signal transduction pathways is well characterized in E. coli, but important details are still not understood. Only recently has a source of gain in the signal transduction network been identified experimentally, and here we present a mathematical model based on dynamic assembly of receptor teams that can explain this observation. PMID- 15111387 TI - Release currents of IP(3) receptor channel clusters and concentration profiles. AB - We simulate currents and concentration profiles generated by Ca(2+) release from the endoplasmic reticulum (ER) to the cytosol through IP(3) receptor channel clusters. Clusters are described as conducting pores in the lumenal membrane with a diameter from 6 nm to 36 nm. The endoplasmic reticulum is modeled as a disc with a radius of 1-12 microm and an inner height of 28 nm. We adapt the dependence of the currents on the trans Ca(2+) concentration (intralumenal) measured in lipid bilayer experiments to the cellular geometry. Simulated currents are compared with signal mass measurements in Xenopus oocytes. We find that release currents depend linearly on the concentration of free Ca(2+) in the lumen. The release current is approximately proportional to the square root of the number of open channels in a cluster. Cytosolic concentrations at the location of the cluster range from 25 microM to 170 microM. Concentration increase due to puffs in a distance of a few micrometers from the puff site is found to be in the nanomolar range. Release currents decay biexponentially with timescales of <1 s and a few seconds. Concentration profiles decay with timescales of 0.125-0.250 s upon termination of release. PMID- 15111388 TI - The facilitated probability of quantal secretion within an array of calcium channels of an active zone at the amphibian neuromuscular junction. AB - A Monte Carlo analysis has been made of the phenomenon of facilitation, whereby a conditioning impulse leaves nerve terminals in a state of heightened release of quanta by a subsequent test impulse, this state persisting for periods of hundreds of milliseconds. It is shown that a quantitative account of facilitation at the amphibian neuromuscular junction can be given if the exocytosis is triggered by the combined action of a low-affinity calcium-binding molecule at the site of exocytosis and a high-affinity calcium-binding molecule some distance away. The kinetic properties and spatial distribution of these molecules at the amphibian neuromuscular junction are arrived at by considering the appropriate values that the relevant parameters must take to successfully account for the experimentally observed amplitude and time course of decline of F1 and F2 facilitation after a conditioning impulse, as well as the growth of facilitation during short trains of impulses. This model of facilitation correctly predicts the effects on facilitation of exogenous buffers such as BAPTA during short trains of impulses. In addition, it accounts for the relative invariance of the kinetics of quantal release due to test-conditioning sequences of impulses as well as due to change in the extent of calcium influx during an impulse. PMID- 15111389 TI - Facilitation through buffer saturation: constraints on endogenous buffering properties. AB - Synaptic facilitation (SF) is a ubiquitous form of short-term plasticity, regulating synaptic dynamics on fast timescales. Although SF is known to depend on the presynaptic accumulation of Ca(2+), its precise mechanism is still under debate. Recently it has been shown that at certain central synapses SF results at least in part from the progressive saturation of an endogenous Ca(2+) buffer (Blatow et al., 2003), as proposed by Klingauf and Neher (1997). Using computer simulations, we study the magnitude of SF that can be achieved by a buffer saturation mechanism (BSM), and explore its dependence on the endogenous buffering properties. We find that a high SF magnitude can be obtained either by a global saturation of a highly mobile buffer in the entire presynaptic terminal, or a local saturation of a completely immobilized buffer. A characteristic feature of BSM in both cases is that SF magnitude depends nonmonotonically on the buffer concentration. In agreement with results of Blatow et al. (2003), we find that SF grows with increasing distance from the Ca(2+) channel cluster, and increases with increasing external Ca(2+), [Ca(2+)](ext), for small levels of [Ca(2+)](ext). We compare our modeling results with the experimental properties of SF at the crayfish neuromuscular junction, and find that the saturation of an endogenous mobile buffer can explain the observed SF magnitude and its supralinear accumulation time course. However, we show that the BSM predicts slowing of the SF decay rate in the presence of exogenous Ca(2+) buffers, contrary to experimental observations at the crayfish neuromuscular junction. Further modeling and data are required to resolve this aspect of the BSM. PMID- 15111390 TI - Size effects on diffusion processes within agarose gels. AB - To investigate diffusion processes in agarose gel, nanoparticles with sizes in the range between 1 and 140 nm have been tested by means of fluorescence correlation spectroscopy. Understanding the diffusion properties in agarose gels is interesting, because such gels are good models for microbial biofilms and cells cytoplasm. The fluorescence correlation spectroscopy technique is very useful for such investigations due to its high sensitivity and selectivity, its excellent spatial resolution compared to the pore size of the gel, and its ability to probe a wide range of sizes of diffusing nanoparticles. The largest hydrodynamic radius (R(c)) of trapped particles that displayed local mobility was estimated to be 70 nm for a 1.5% agarose gel. The results showed that diffusion of particles in agarose gel is anomalous, with a diverging fractal dimension of diffusion when the large particles become entrapped in the pores of the gel. The latter situation occurs when the reduced size (R(A)/R(c)) of the diffusing particle, A, is >0.4. Variations of the fractal exponent of diffusion (d(w)) with the reduced particle size were in agreement with three-dimensional Monte Carlo simulations in porous media. Nonetheless, a systematic offset of d(w) was observed in real systems and was attributed to weak nonelastic interactions between the diffusing particles and polymer fibers, which was not considered in the Monte Carlo simulations. PMID- 15111391 TI - A mechanistic model of the actin cycle. AB - We have derived a broad, deterministic model of the steady-state actin cycle that includes its major regulatory mechanisms. Ours is the first model to solve the complete nucleotide profile within filaments, a feature that determines the dynamics and geometry of actin networks at the leading edges of motile cells, and one that has challenged investigators developing models to interpret steady-state experiments. We arrived at the nucleotide profile through analytic and numerical approaches that completely agree. Our model reproduces behaviors seen in numerous experiments with purified proteins, but allows a detailed inspection of the concentrations and fluxes that might exist in these experiments. These inspections provide new insight into the mechanisms that determine the rate of actin filament treadmilling. Specifically, we find that mechanisms for enhancing Pi release from the ADP.Pi intermediate on filaments, for increasing the off rate of ADP-bound subunits at pointed ends, and the multiple, simultaneous functions of profilin, make unique and essential contributions to increased treadmilling. In combination, these mechanisms have a theoretical capacity to increase treadmilling to levels limited only by the amount of available actin. This limitation arises because as the cycle becomes more dynamic, it tends toward the unpolymerized state. PMID- 15111392 TI - Elastic instability in growing yeast colonies. AB - The differential adhesion between cells is believed to be the major driving force behind the formation of tissues. The idea is that an aggregate of cells minimizes the overall adhesive energy between cell surfaces. We demonstrate in a model experimental system that there exist conditions where a slowly growing tissue does not minimize this adhesive energy. A mathematical model demonstrates that the instability of a spherical shape is caused by the competition between elastic and surface energies. PMID- 15111393 TI - The sonic hedgehog signaling system as a bistable genetic switch. AB - Sonic hedgehog (Shh) controls critical cellular decisions between distinct fates in many systems, particularly in stem cells. The Shh network functions as a genetic switch, and we have theoretically and computationally analyzed how its structure can endow it with the ability to switch fate choices at a threshold Shh concentration. The network is composed of a positive transcriptional feedback loop embedded within a negative signaling feedback loop. Specifically, positive feedback by the transcription factor Gli, which upregulates its own expression, leads to a switch that can adopt two distinct states as a function of Shh. However, Gli also upregulates the signaling repressor Patched, negative feedback that reins in the strong Gli autoregulatory loop. Mutations that have been associated with cancer are predicted to yield an irreversible switch to a high Gli state. Finally, stochastic simulation reveals the negative Patched feedback loop serves a critical function of dampening Gli fluctuations to reduce spontaneous state switching and preserve the network's robust, switch-like behavior. Tightly linked positive and negative feedback loops are present in many signaling systems, and the Shh system is therefore likely representative of a large set of gene regulation networks that control stem cell fate throughout development and into adulthood. PMID- 15111394 TI - Stability and the evolvability of function in a model protein. AB - Functional proteins must fold with some minimal stability to a structure that can perform a biochemical task. Here we use a simple model to investigate the relationship between the stability requirement and the capacity of a protein to evolve the function of binding to a ligand. Although our model contains no built in tradeoff between stability and function, proteins evolved function more efficiently when the stability requirement was relaxed. Proteins with both high stability and high function evolved more efficiently when the stability requirement was gradually increased than when there was constant selection for high stability. These results show that in our model, the evolution of function is enhanced by allowing proteins to explore sequences corresponding to marginally stable structures, and that it is easier to improve stability while maintaining high function than to improve function while maintaining high stability. Our model also demonstrates that even in the absence of a fundamental biophysical tradeoff between stability and function, the speed with which function can evolve is limited by the stability requirement imposed on the protein. PMID- 15111395 TI - Atomic mean-square displacements in proteins by molecular dynamics: a case for analysis of variance. AB - Information on protein internal motions is usually obtained through the analysis of atomic mean-square displacements, which are a measure of variability of the atomic positions distribution functions. We report a statistical approach to analyze molecular dynamics data on these displacements that is based on probability distribution functions. Using a technique inspired by the analysis of variance, we compute unbiased, reliable mean-square displacements of the atoms and analyze them statistically. We applied this procedure to characterize protein thermostability by comparing the results for a thermophilic enzyme and a mesophilic homolog. In agreement with previous experimental observations, our analysis suggests that the proteins surface regions can play a role in the different thermal behavior. PMID- 15111396 TI - Modeling electron transfer thermodynamics in protein complexes: interaction between two cytochromes c(3). AB - Redox protein complexes between type I and type II tetraheme cytochromes c(3) from Desulfovibrio vulgaris Hildenborough are here analyzed using theoretical methodologies. Various complexes were generated using rigid-body docking techniques, and the two lowest energy complexes (1 and 2) were relaxed using molecular dynamics simulations with explicit solvent and subjected to further characterization. Complex 1 corresponds to an interaction between hemes I from both cytochromes c(3). Complex 2 corresponds to an interaction between the heme IV from type I and the heme I from type II cytochrome c(3). Binding free energy calculations using molecular mechanics, Poisson-Boltzmann, and surface accessibility methods show that complex 2 is more stable than complex 1. Thermodynamic calculations on complex 2 show that complex formation induces changes in the reduction potential of both cytochromes c(3), but the changes are larger in the type I cytochrome c(3) (the largest one occurring on heme IV, of approximately 80 mV). These changes are sufficient to invert the global titration curves of both cytochromes, generating directionally in electron transfer from type I to type II cytochrome c(3), a phenomenon of obvious thermodynamic origin and consequences, but also with kinetic implications. The existence of processes like this occurring at complex formation may constitute a natural design of efficient redox chains. PMID- 15111397 TI - Simulation of Drosophila circadian oscillations, mutations, and light responses by a model with VRI, PDP-1, and CLK. AB - A model of Drosophila circadian rhythm generation was developed to represent feedback loops based on transcriptional regulation of per, Clk (dclock), Pdp-1, and vri (vrille). The model postulates that histone acetylation kinetics make transcriptional activation a nonlinear function of [CLK]. Such a nonlinearity is essential to simulate robust circadian oscillations of transcription in our model and in previous models. Simulations suggest that two positive feedback loops involving Clk are not essential for oscillations, because oscillations of [PER] were preserved when Clk, vri, or Pdp-1 expression was fixed. However, eliminating positive feedback by fixing vri expression altered the oscillation period. Eliminating the negative feedback loop in which PER represses per expression abolished oscillations. Simulations of per or Clk null mutations, of per overexpression, and of vri, Clk, or Pdp-1 heterozygous null mutations altered model behavior in ways similar to experimental data. The model simulated a photic phase-response curve resembling experimental curves, and oscillations entrained to simulated light-dark cycles. Temperature compensation of oscillation period could be simulated if temperature elevation slowed PER nuclear entry or PER phosphorylation. The model makes experimental predictions, some of which could be tested in transgenic Drosophila. PMID- 15111398 TI - Radial arrangement of chromosome territories in human cell nuclei: a computer model approach based on gene density indicates a probabilistic global positioning code. AB - Numerous investigations in the last years focused on chromosome arrangements in interphase nuclei. Recent experiments concerning the radial positioning of chromosomes in the nuclear volume of human and primate lymphocyte cells suggest a relationship between the gene density of a chromosome territory (CT) and its distance to the nuclear center. To relate chromosome positioning and gene density in a quantitative way, computer simulations of whole human cell nuclear genomes of normal karyotype were performed on the basis of the spherical 1 Mbp chromatin domain model and the latest data about sequence length and gene density of chromosomes. Three different basic assumptions about the initial distribution of chromosomes were used: a statistical, a deterministic, and a probabilistic initial distribution. After a simulated decondensation in early G1, a comparison of the radial distributions of simulated and experimentally obtained data for CTs Nos. 12, 18, 19, and 20 was made. It was shown that the experimentally observed distributions can be fitted better assuming an initial probabilistic distribution. This supports the concept of a probabilistic global gene positioning code depending on CT sequence length and gene density. PMID- 15111399 TI - Model of creation and evolution of stable electropores for DNA delivery. AB - Electroporation, in which electric pulses create transient pores in the cell membrane, is becoming an important technique for gene therapy. To enable entry of supercoiled DNA into cells, the pores should have sufficiently large radii (>10 nm), remain open long enough for the DNA chain to enter the cell (milliseconds), and should not cause membrane rupture. This study presents a model that can predict such macropores. The distinctive features of this model are the coupling of individual pores through membrane tension and the electrical force on the pores, which is applicable to pores of any size. The model is used to explore the process of pore creation and evolution and to determine the number and size of pores as a function of the pulse magnitude and duration. Next, our electroporation model is combined with a heuristic model of DNA uptake and used to predict the dependence of DNA uptake on pulsing parameters. Finally, the model is used to examine the mechanism of a two-pulse protocol, which was proposed specifically for gene delivery. The comparison between experimental results and the model suggests that this model is well-suited for the investigation of electroporation-mediated DNA delivery. PMID- 15111400 TI - Water alignment, dipolar interactions, and multiple proton occupancy during water wire proton transport. AB - A discrete multistate kinetic model for water-wire proton transport is constructed and analyzed using Monte Carlo simulations. In the model, each water molecule can be in one of three states: oxygen lone-pairs pointing leftward, pointing rightward, or protonated (H(3)O(+)). Specific rules for transitions among these states are defined as protons hop across successive water oxygens. Our model also includes water-channel interactions that preferentially align the water dipoles, nearest-neighbor dipolar coupling interactions, and Coulombic repulsion. Extensive Monte Carlo simulations were performed and the observed qualitative physical behaviors discussed. We find the parameters that allow the model to exhibit superlinear and sublinear current-voltage relationships, and show why alignment fields, whether generated by interactions with the pore interior or by membrane potentials, always decrease the proton current. The simulations also reveal a "lubrication" mechanism that suppresses water dipole interactions when the channel is multiply occupied by protons. This effect can account for an observed sublinear-to-superlinear transition in the current voltage relationship. PMID- 15111401 TI - Structural restraints and heterogeneous orientation of the gramicidin A channel closed state in lipid bilayers. AB - Although there have been several decades of literature illustrating the opening and closing of the monovalent cation selective gramicidin A channel through single channel conductance, the closed conformation has remained poorly characterized. In sharp contrast, the open-state dimer is one of the highest resolution structures yet characterized in a lipid environment. To shift the open/closed equilibrium dramatically toward the closed state, a lower peptide/lipid molar ratio and, most importantly, long-chain lipids have been used. For the first time, structural evidence for a monomeric state has been observed for the native gramicidin A peptide. Solid-state NMR spectroscopy of single-site (15)N-labeled gramicidin in uniformly aligned bilayers in the L(alpha) phase have been observed. The results suggest a kinked structure with considerable orientational heterogeneity. The C-terminal domain is well structured, has a well-defined orientation in the bilayer, and appears to be in the bilayer interfacial region. On the other hand, the N-terminal domain, although appearing to be well structured and in the hydrophobic core of the bilayer, has a broad range of orientations relative to the bilayer normal. The structure is not just half of the open-state dimer, and neither is the structure restricted to the surface of the bilayer. Consequently, the monomeric or closed state appears to be a hybrid of these two models from the literature. PMID- 15111402 TI - Gating of the large mechanosensitive channel in situ: estimation of the spatial scale of the transition from channel population responses. AB - Physical expansion associated with the opening of a tension-sensitive channel has the same meaning as gating charge for a voltage-gated channel. Despite increasing evidence for the open-state conformation of MscL, the energetic description of its complex gating remains incomplete. The previously estimated in-plane expansion of MscL is considerably smaller than predicted by molecular models. To resolve this discrepancy, we conducted a systematic study of currents and dose response curves for wild-type MscL in Escherichia coli giant spheroplasts. Using the all-point histogram method and calibrating tension against the threshold for the small mechanosensitive channel (MscS) in each patch, we found that the distribution of channels among the subconducting states is significantly less dependent on tension than the distribution between the closed and conducting states. At -20 mV, all substates together occupy approximately 30% of the open time and reduce the mean integral current by approximately 6%, essentially independent of tension or P(o). This is consistent with the gating scheme in which the major rate-limiting step is the transition between the closed state and a low-conducting substate, and validates both the use of the integral current as a measure of P(o), and treatment of dose-response curves in the two-state approximation. The apparent energy and area differences between the states deltaE and deltaA, extracted from 29 independent dose-response curves, varied in a linearly correlated manner whereas the midpoint tension stayed at approximately 10.4 mN/m. Statistical modeling suggests slight variability of gating parameters among channels in each patch, causing a strong reduction and correlated spread of apparent deltaE and deltaA. The slope of initial parts of activation curves, with a few channels being active, gave estimates of deltaE = 51 +/- 13 kT and deltaA = 20.4 +/- 4.8 nm(2), the latter being consistent with structural models of MscL, which predict deltaA = 23 nm(2). PMID- 15111403 TI - Cysteine scanning of MscL transmembrane domains reveals residues critical for mechanosensitive channel gating. AB - The mechanosensitive channel of large conductance (MscL), a bacterial channel, is perhaps the best characterized mechanosensitive protein. A structure of the Mycobacterium tuberculosis ortholog has been solved by x-ray crystallography, but details of how the channel gates remain obscure. Here, cysteine scanning was used to identify residues within the transmembrane domains of Escherichia coli MscL that are crucial for normal function. Utilizing genetic screens, we identified several mutations that induced gain-of-function or loss-of-function phenotypes in vivo. Mutants that exhibited the most severe phenotypes were further characterized using electrophysiological techniques and chemical modifications of the substituted cysteines. Our results verify the importance of residues in the putative primary gate in the first transmembrane domain, corroborate other residues previously noted as critical for normal function, and identify new ones. In addition, evaluation of disulfide bridging in native membranes suggests alterations of existing structural models for the "fully closed" state of the channel. PMID- 15111404 TI - Functional effects of auxiliary beta4-subunit on rat large-conductance Ca(2+) activated K(+) channel. AB - Large-conductance calcium-activated potassium (BK(Ca)) channels are composed of the pore-forming alpha-subunit and the auxiliary beta-subunits. The beta4-subunit is dominantly expressed in the mammalian central nervous system. To understand the physiological roles of the beta4-subunit on the BK(Ca) channel alpha-subunit (Slo), we isolated a full-length complementary DNA of rat beta4-subunit (rbeta4), expressed heterolgously in Xenopus oocytes, and investigated the detailed functional effects using electrophysiological means. When expressed together with rat Slo (rSlo), rbeta4 profoundly altered the gating characteristics of the Slo channel. At a given concentration of intracellular Ca(2+), rSlo/rbeta4 channels were more sensitive to transmembrane voltage changes. The activation and deactivation rates of macroscopic currents were decreased in a Ca(2+)-dependent manner. The channel activation by Ca(2+) became more cooperative by the coexpression of rbeta4. Single-channel recordings showed that the increased Hill coefficient for Ca(2+) was due to the changes in the open probability of the rSlo/rbeta4 channel. Single BK(Ca) channels composed of rSlo and rbeta4 also exhibited slower kinetics for steady-state gating compared with rSlo channels. Dwell times of both open and closed events were significantly increased. Because BK(Ca) channels are known to modulate neuroexcitability and the expression of the beta4-subunit is highly concentrated in certain subregions of brain, the electrophysiological properties of individual neurons should be affected profoundly by the expression of this second subunit. PMID- 15111405 TI - Water dynamics and dewetting transitions in the small mechanosensitive channel MscS. AB - The dynamics of confined water in capillaries and nanotubes suggests that gating of ion channels may involve not only changes of the pore geometry, but also transitions between water-filled and empty states in certain locations. The recently solved heptameric structure of the small mechanosensitive channel of Escherichia coli, MscS, has revealed a relatively wide (7-15 A) yet highly hydrophobic transmembrane pore. Continuum estimations based on the properties of pore surface suggest low conductance and a thermodynamic possibility of dewetting. To test the predictions we performed molecular dynamics simulations of MscS filled with flexible TIP3P water. Irrespective to the initial conditions, several independent 6-ns simulations converged to the same stable state with the pore water-filled in the wider part, but predominantly empty in the narrow hydrophobic part, displaying intermittent vapor-liquid transitions. The polar gain-of-function substitution L109S in the constriction resulted in a stable hydration of the entire pore. Steered passages of Cl(-) ions through the narrow part of the pore consistently produced partial ion dehydration and required a force of 200-400 pN to overcome an estimated barrier of 10-20 kcal/mole, implying negligibly low conductance. We conclude that the crystal structure of MscS does not represent an open state. We infer that MscS gate, which is similar to that of the nicotinic ACh receptor, involves a vapor-lock mechanism where limited changes of geometry or surface polarity can locally switch the regime between water filled (conducting) and empty (nonconducting) states. PMID- 15111406 TI - Short class I major histocompatibility complex cytoplasmic tails differing in charge detect arbiters of lateral diffusion in the plasma membrane. AB - Directed and Brownian movement of class I major histocompatibility complex (MHC) molecules on cell membranes is implicated in antigen presentation. Previous studies indicated that the class I MHC cytoplasmic tail imposes constraints on the molecule's diffusion. Here we used single particle tracking to study the mobility of the wild-type mouse H-2L(d) class I MHC molecule and of seven cytoplasmic tail variants. Six of the variants have cytoplasmic tails of four or seven residues (differing in net charge), and one is tailless, yet all are susceptible to confinement in membrane domains. However, truncation of the cytoplasmic tail to 0-4 residues decreases the proportion of particles exhibiting confined diffusion and increases the proportion exhibiting simple diffusion. Particularly for the truncated mutants (tail length of 0-7 residues), many of the particles have complex trajectories and do not move at a constant speed or in the same mode of diffusion throughout the observation period. Several particles of the tailless H-2L(d) mutant display a type of directed diffusion that is rarely observed for other H-2L(d) mutants. Taken together, these data show that even short cytoplasmic tails can influence markedly class I MHC mobility and that cytoplasmic tail length and sequence affect the molecule's diffusion in the membrane. PMID- 15111407 TI - Liquid domains in vesicles investigated by NMR and fluorescence microscopy. AB - We use (2)H-NMR, (1)H-MAS NMR, and fluorescence microscopy to detect immiscibility in three particular phospholipid ratios mixed with 30% cholesterol: 2:1 DOPC/DPPC, 1:1 DOPC/DPPC, and 1:2 DOPC/DPPC. Large-scale (>>160 nm) phase separation into liquid-ordered (L(o)) and liquid-crystalline (L(alpha)) phases is observed by both NMR and fluorescence microscopy. By fitting superimposed (2)H NMR spectra, we quantitatively determine that the L(o) phase is strongly enriched in DPPC and moderately enriched in cholesterol. Tie-lines estimated at different temperatures and membrane compositions are based on both (2)H-NMR observations and a previously published ternary phase diagram. (2)H- and (1)H-MAS NMR techniques probe significantly smaller length scales than microscopy experiments (submicron versus micron-scalp), and complex behavior is observed near the miscibility transition. Fluorescence microscopy of giant unilamellar vesicles shows micrometer-scale domains below the miscibility transition. In contrast, NMR of multilamellar vesicles gives evidence for smaller ( approximately 80 nm) domains just below the miscibility transition, whereas large-scale demixing occurs at a lower temperature, T(low). A transition at T(low) is also evident in fluorescence microscopy measurements of the surface area fraction of ordered phase in giant unilamellar vesicles. Our results reemphasize the complex phase behavior of cholesterol-containing membranes and provide a framework for interpreting (2)H-NMR experiments in similar membranes. PMID- 15111408 TI - Tension in tubulovesicular networks of Golgi and endoplasmic reticulum membranes. AB - The endoplasmic reticulum (ER) and Golgi have robust bidirectional traffic between them and yet form distinct membrane compartments. Membrane tubules are pulled from large aggregates of ER or Golgi by microtubule motors to form ER tubulovesicular networks or Golgi tubules both in vivo and in vitro. The physical properties of membranes are critical for membrane traffic and organelle morphology. For example, tension applied to membranes can create tethers, drive membrane flow, and set the diameter of the tubules. Here, we formed ER and Golgi membrane networks in vitro and used optical tweezers to measure directly, for the first time, the membrane tensions of these organelles to clarify the possible role of tension in membrane flow. We report that higher forces are needed to form tethers from ER (18.6 +/- 2.8 pN) than from Golgi (11.4 +/- 1.4 pN) membrane tubules in vitro. Since ER tubules are smaller in diameter than Golgi tubules, it follows that Golgi networks have a lower tension than ER. The lower tension of the ER could be an explanation of how Golgi tubules can be rapidly drawn into the ER by tension-driven flow after fusion, as is observed in vivo. PMID- 15111409 TI - Bimodal distribution and fluorescence response of environment-sensitive probes in lipid bilayers. AB - A remarkable heterogeneity is often observed in the spectroscopic properties of environment-sensitive fluorescence probes in phospholipid bilayers. To explain its origin, we provided a detailed investigation of the fluorescence excitation and emission spectra of 4'-dimethylamino-3-hydroxyflavone (probe F) in bilayer vesicles with the variations of fatty acid composition, polar heads, temperature, and cholesterol content. Probe F, due to excited-state intramolecular proton transfer, exhibits two bands in emission that are differently sensitive to intermolecular interactions-thereby allowing us to distinguish universal (dipole dipole) and specific (H-bonding) interactions within the bilayer. Spectroscopic, quenching, and anisotropy data suggest the presence of two forms of probe F at different locations in the bilayer: an H-bond free form located below sn(1) carbonyls and an H-bonded form located at the polar membrane interface. We provide a quantitative analysis of the distribution of the probe between these two locations as well as the polarity of these locations, and show that both the distribution and the polarity contribute to the probe response. Moreover, analysis of literature data on other environment-sensitive probes (Prodan, Laurdan, Nile Red, NBD lipids, etc.) in lipid bilayers allows us to suggest that the bimodal distribution in the lipid bilayer is probably a general feature of low-polar molecules with polar groups capable of H-bonding interactions. PMID- 15111410 TI - Nonequilibrium behavior in supported lipid membranes containing cholesterol. AB - We investigate lateral organization of lipid domains in vesicles versus supported membranes and monolayers. The lipid mixtures used are predominantly DOPC/DPPC/Chol and DOPC/BSM/Chol, which have been previously shown to produce coexisting liquid phases in vesicles and monolayers. In a monolayer at an air water interface, these lipids have miscibility transition pressures of approximately 12-15 mN/m, which can rise to 32 mN/m if the monolayer is exposed to air. Lipid monolayers can be transferred by Langmuir-Schafer deposition onto either silanized glass or existing Langmuir-Blodgett supported monolayers. Micron scale domains are present in the transferred lipids only if they were present in the original monolayer before deposition. This result is valid for transfers at 32 mN/m and also at lower pressures. Domains transferred to glass supports differ from liquid domains in vesicles because they are static, do not align in registration across leaflets, and do not reappear after temperature is cycled. Similar static domains are found for vesicles ruptured onto glass surfaces. Although supported membranes on glass capture some aspects of vesicles in equilibrium (e.g., gel-liquid transition temperatures and diffusion rates of individual lipids), the collective behavior of lipids in large liquid domains is poorly reproduced. PMID- 15111411 TI - Energetics of vesicle fusion intermediates: comparison of calculations with observed effects of osmotic and curvature stresses. AB - We reported previously the effects of both osmotic and curvature stress on fusion between poly(ethylene glycol)-aggregated vesicles. In this article, we analyze the energetics of fusion of vesicles of different curvature, paying particular attention to the effects of osmotic stress on small, highly curved vesicles of 26 nm diameter, composed of lipids with negative intrinsic curvature. Our calculations show that high positive curvature of the outer monolayer "charges" these vesicles with excess bending energy, which then releases during stalk expansion (increase of the stalk radius, r(s)) and thus "drives" fusion. Calculations based on the known mechanical properties of lipid assemblies suggest that the free energy of "void" formation as well as membrane-bending free energy dominate the evolution of a stalk to an extended transmembrane contact. The free energy profile of stalk expansion (free energy versus r(s)) clearly shows the presence of two metastable intermediates (intermediate 1 at r(s) approximately 0 1.0 nm and intermediate 2 at r(s) approximately 2.5 - 3.0 nm). Applying osmotic gradients of +/-5 atm, when assuming a fixed trans-bilayer lipid mass distribution, did not significantly change the free-energy profile. However, inclusion in the model of an additional degree of freedom, the ability of lipids to move into and out of the "void", made the free-energy profile strongly dependent on the osmotic gradient. Vesicle expansion increased the energy barrier between intermediates by approximately 4 kT and the absolute value of the barrier by approximately 7 kT, whereas compression decreased it by nearly the same extent. Since these calculations, which are based on the stalk hypothesis, correctly predict the effects of both membrane curvature and osmotic stress, they support the stalk hypothesis for the mechanism of membrane fusion and suggest that both forms of stress alter the final stages, rather than the initial step, of the fusion process, as previously suggested. PMID- 15111412 TI - Role of cholesterol in the formation and nature of lipid rafts in planar and spherical model membranes. AB - Sterols play a crucial regulatory and structural role in the lateral organization of eukaryotic cell membranes. Cholesterol has been connected to the possible formation of ordered lipid domains (rafts) in mammalian cell membranes. Lipid rafts are composed of lipids in the liquid-ordered (l(o)) phase and are surrounded with lipids in the liquid-disordered (l(d)) phase. Cholesterol and sphingomyelin are thought to be the principal components of lipid rafts in cell and model membranes. We have used fluorescence microscopy and fluorescence recovery after photobleaching in planar supported lipid bilayers composed of porcine brain phosphatidylcholine (bPC), porcine brain sphingomyelin (bSM), and cholesterol to map the composition-dependence of l(d)/l(o) phase coexistence. Cholesterol decreases the fluidity of bPC bilayers, but disrupts the highly ordered gel phase of bSM, leading to a more fluid membrane. When mixed with bPC/bSM (1:1) or bPC/bSM (2:1), cholesterol induces the formation of l(o) phase domains. The fraction of the membrane in the l(o) phase was found to be directly proportional to the cholesterol concentration in both phospholipid mixtures, which implies that a significant fraction of bPC cosegregates into l(o) phase domains. Images reveal a percolation threshold, i.e., the point where rafts become connected and fluid domains disconnected, when 45-50% of the total membrane is converted to the l(o) phase. This happens between 20 and 25 mol % cholesterol in 1:1 bPC/bSM bilayers and between 25 and 30 mol % cholesterol in 2:1 bPC/bSM bilayers at room temperature, and at approximately 35 mol % cholesterol in 1:1 bPC/bSM bilayers at 37 degrees C. Area fractions of l(o) phase lipids obtained in multilamellar liposomes by a fluorescence resonance energy transfer method confirm and support the results obtained in planar lipid bilayers. PMID- 15111413 TI - Domain formation in phosphatidylinositol monophosphate/phosphatidylcholine mixed vesicles. AB - Phosphoinositides have been shown to control membrane trafficking events by targeting proteins to specific cellular sites, which requires a tight regulation of phosphoinositide generation and turnover as well as a high degree of compartmentalization. To shed light on the processes that lead to the formation of phosphoinositide-enriched microdomains, phosphatidylcholine/phosphatidylinositol monophosphate (phosphatidylinositol-3 phosphate (PI-3P), -4-phosphate (PI-4P), or -5-phosphate (PI-5P)) mixed vesicles were investigated by calorimetric (DSC) Fourier transform infrared spectroscopic (FTIR), and fluorescence resonance energy transfer (FRET) measurements. The experiments furnished results consistent with a pH-dependent formation of phosphatidylinositol monophosphate-enriched microdomains. The domain formation was most pronounced between pH approximately 7 and approximately 9.5, whereas slightly acidic pH values (pH 4) resulted in the disintegration of the domains. This pH-dependent phosphatidylcholine/phosphatidylinositol monophosphate demixing was observed for the gel phase (FTIR experiments) as well as for the fluid lipid phase (FRET measurements). The observed microdomains are presumably stabilized by hydroxyl/hydroxyl as well as hydroxyl/phosphomonoester and phosphodiester interactions. While the pH dependence of the mutual phosphatidylinositol monophosphate interaction was largely the same for all investigated phosphatidylinositol monophosphates, it turned out that the relative stability of phosphatidylinositol monophosphate-enriched microdomains (pH 7-9.5) was governed by the position of the phosphomonoester group at the inositol ring (PI-4P > PI-5P > PI-3P). Demixing was also observed for phosphatidylcholine/phosphatidylinositol mixed vesicles; however, in this case the microdomain formation was only slightly affected by pH changes. PMID- 15111414 TI - Structural and functional roles of desmin in mouse skeletal muscle during passive deformation. AB - Mechanical interactions between desmin and Z-disks, costameres, and nuclei were measured during passive deformation of single muscle cells. Image processing and continuum kinematics were used to quantify the structural connectivity among these structures. Analysis of both wild-type and desmin-null fibers revealed that the costamere protein talin colocalized with the Z-disk protein alpha-actinin, even at very high strains and stresses. These data indicate that desmin is not essential for mechanical coupling of the costamere complex and the sarcomere lattice. Within the sarcomere lattice, significant differences in myofibrillar connectivity were revealed between passively deformed wild-type and desmin-null fibers. Connectivity in wild-type fibers was significantly greater compared to desmin-null fibers, demonstrating a significant functional connection between myofibrils that requires desmin. Passive mechanical analysis revealed that desmin may be partially responsible for regulating fiber volume, and consequently, fiber mechanical properties. Kinematic analysis of alpha-actinin strain fields revealed that knockout fibers transmitted less shear strain compared to wild-type fibers and experienced a slight increase in fiber volume. Finally, linkage of desmin intermediate filaments to muscle nuclei was strongly suggested based on extensive loss of nuclei positioning in the absence of desmin during passive fiber loading. PMID- 15111415 TI - Cross-bridge number, position, and angle in target zones of cryofixed isometrically active insect flight muscle. AB - Electron micrographic tomograms of isometrically active insect flight muscle, freeze substituted after rapid freezing, show binding of single myosin heads at varying angles that is largely restricted to actin target zones every 38.7 nm. To quantify the parameters that govern this pattern, we measured the number and position of attached myosin heads by tracing cross-bridges through the three dimensional tomogram from their origins on 14.5-nm-spaced shelves along the thick filament to their thin filament attachments in the target zones. The relationship between the probability of cross-bridge formation and axial offset between the shelf and target zone center was well fitted by a Gaussian distribution. One head of each myosin whose origin is close to an actin target zone forms a cross-bridge most of the time. The probability of cross-bridge formation remains high for myosin heads originating within 8 nm axially of the target zone center and is low outside 12 nm. We infer that most target zone cross-bridges are nearly perpendicular to the filaments (60% within 11 degrees ). The results suggest that in isometric contraction, most cross-bridges maintain tension near the beginning of their working stroke at angles near perpendicular to the filament axis. Moreover, in the absence of filament sliding, cross-bridges cannot change tilt angle while attached nor reach other target zones while detached, so may cycle repeatedly on and off the same actin target monomer. PMID- 15111416 TI - Fluorescence depolarization of actin filaments in reconstructed myofibers: the effect of S1 or pPDM-S1 on movements of distinct areas of actin. AB - Fluorescence polarization measurements were used to study changes in the orientation and order of different sites on actin monomers within muscle thin filaments during weak or strong binding states with myosin subfragment-1. Ghost muscle fibers were supplemented with actin monomers specifically labeled with different fluorescent probes at Cys-10, Gln-41, Lys-61, Lys-373, Cys-374, and the nucleotide binding site. We also used fluorescent phalloidin as a probe near the filament axis. Changes in the orientation of the fluorophores depend not only on the state of acto-myosin binding but also on the location of the fluorescent probes. We observed changes in polarization (i.e., orientation) for those fluorophores attached at the sites directly involved in myosin binding (and located at high radii from the filament axis) that were contrary to the fluorophores located at the sites close to the axis of thin filament. These altered probe orientations suggest that myosin binding alters the conformation of F-actin. Strong binding by myosin heads produces changes in probe orientation that are opposite to those observed during weak binding. PMID- 15111417 TI - Myosin regulatory domain orientation in skeletal muscle fibers: application of novel electron paramagnetic resonance spectral decomposition and molecular modeling methods. AB - Reorientation of the regulatory domain of the myosin head is a feature of all current models of force generation in muscle. We have determined the orientation of the myosin regulatory light chain (RLC) using a spin-label bound rigidly and stereospecifically to the single Cys-154 of a mutant skeletal isoform. Labeled RLC was reconstituted into skeletal muscle fibers using a modified method that results in near-stoichiometric levels of RLC and fully functional muscle. Complex electron paramagnetic resonance spectra obtained in rigor necessitated the development of a novel decomposition technique. The strength of this method is that no specific model for a complex orientational distribution was presumed. The global analysis of a series of spectra, from fibers tilted with respect to the magnetic field, revealed two populations: one well-ordered (+/-15 degrees ) with the spin-label z axis parallel to actin, and a second population with a large distribution (+/-60 degrees ). A lack of order in relaxed or nonoverlap fibers demonstrated that regulatory domain ordering was defined by interaction with actin rather than the thick filament surface. No order was observed in the regulatory domain during isometric contraction, consistent with the substantial reorientation that occurs during force generation. For the first time, spin-label orientation has been interpreted in terms of the orientation of a labeled domain. A Monte Carlo conformational search technique was used to determine the orientation of the spin-label with respect to the protein. This in turn allows determination of the absolute orientation of the regulatory domain with respect to the actin axis. The comparison with the electron microscopy reconstructions verified the accuracy of the method; the electron paramagnetic resonance determined that axial orientation was within 10 degrees of the electron microscopy model. PMID- 15111419 TI - Tracking operator state fluctuations in gene expression in single cells. AB - We report the results of operator state fluctuations in gene expression for the entire bacterial growth cycle, using single-cell analysis and synthetic unregulated and negative-feedback transcription regulatory gene circuits. In the unregulated circuit, during the cell cycle, we observe a crossover from log normal-to-normal distribution of expressed proteins and an unusual linear dependence of their standard deviation on the mean gene expression levels. With negative-feedback circuits we find the existence of bimodality as the cell cycle progresses. We suggest that such long-tail and bimodal distributions may be used as selection mechanisms in developmental switches and for assigning cell identity. PMID- 15111418 TI - Slow calcium signals after tetanic electrical stimulation in skeletal myotubes. AB - The fluorescent calcium signal from rat myotubes in culture was monitored after field-stimulation with tetanic protocols. After the calcium signal sensitive to ryanodine and associated to the excitation-contraction coupling, a second long lasting calcium signal refractory to ryanodine was consistently found. The onset kinetics of this slow signal were slightly modified in nominally calcium-free medium, as were both the frequency and number of pulses during tetanus. No signal was detected in the presence of tetrodotoxin. The participation of the dihydropyridine receptor (DHPR) as the voltage sensor for this signal was assessed by treatment with agonist and antagonist dihydropyridines (Bay K 8644 and nifedipine), showing an enhanced and inhibitory response, respectively. In the dysgenic GLT cell line, which lacks the alpha1(S) subunit of the DHPR, the signal was absent. Transfection of these cells with the alpha1(S) subunit restored the slow signal. In myotubes, the inositol 1,4,5-trisphosphate (IP(3)) mass increase induced by a tetanus protocol preceded in time the slow calcium signal. Both an IP(3) receptor blocker and a phospholipase C inhibitor (xestospongin C and U73122, respectively) dramatically inhibit this signal. Long lasting, IP(3)-generated slow calcium signals appear to be a physiological response to activity-related fluctuations in membrane potential sensed by the DHPR. PMID- 15111420 TI - Low-resolution reconstruction of a synthetic DNA holliday junction. AB - We have studied the low-resolution solution conformation of a Holliday (or four way) DNA junction by using small-angle x-ray scattering, sedimentation velocity, and computational modeling techniques. The scattering data were analyzed in two independent ways: firstly, by rigid-body modeling of the scattering data using previously suggested models for the Holliday junction (HJ), and secondly, by ab initio reconstruction methods. The models found by both methods agree with experimentally determined sedimentation coefficients and are compatible with the results of previous studies using different techniques, but provide a more direct and accurate determination of the solution conformation of the HJ. Our results show that addition of Mg(2+) alters the conformation of the HJ from an extended to a stacked arrangement. PMID- 15111421 TI - Specific versus nonspecific binding of cationic PNAs to duplex DNA. AB - Although peptide nucleic acids (PNAs) are neutral by themselves, they are usually appended with positively charged lysine residues to increase their solubility and binding affinity for nucleic acid targets. Thus obtained cationic PNAs very effectively interact with the designated duplex DNA targets in a sequence specific manner forming strand-invasion complexes. We report on the study of the nonspecific effects in the kinetics of formation of sequence-specific PNA-DNA complexes. We find that in a typical range of salt concentrations used when working with strand-invading PNAs (10-20 mM NaCl) the PNA binding rates essentially do not depend on the presence of nontarget DNA in the reaction mixture. However, at lower salt concentrations (<10 mM NaCl), the rates of PNA binding to DNA targets are significantly slowed down by the excess of unrelated DNA. This effect of nontarget DNA arises from depleting the concentration of free PNA capable of interacting with DNA target due to adhesion of positively charged PNA molecules on the negatively charged DNA duplex. As expected, the nonspecific electrostatic effects are more pronounced for more charged PNAs. We propose a simple model quantitatively describing all major features of the observed phenomenon. This understanding is important for design of and manipulation with the DNA-binding polycationic ligands in general and PNA-based drugs in particular. PMID- 15111422 TI - Monte Carlo simulations of locally melted supercoiled DNAs in 20 mM ionic strength. AB - Mesoscopic models of unmelted and locally melted supercoiled DNAs in 20 mM ionic strength are simulated over a range of linking difference from deltal = 0 to -26 turns, or superhelix density from sigma = 0 to -0.062. A domain containing m = 0, 28, or 56 melted basepairs (out of 4349 total) is modeled simply by a region of suitable length with substantially reduced torsion and bending elastic constants. Average structural properties are calculated from the saved configurations, and a reversible work protocol is used to calculate the supercoiling free energy, The cross-writhe between duplex and melted regions (defined herein) is found to be negligibly small. The total writhe, radius of gyration, and ordered elements of the diagonalized inertial tensor are found to be nearly universal functions of the residual linking difference (deltal(r)) associated with the duplex region, independent of m. However, deformability of the tertiary structure, as manifested by the variance of those same properties, is not a universal function of deltal(r)), but depends upon m.delta (SC) varies with deltal(r)) more strongly than deltal(r)) (2)due to the low ionic strength. The twist energy parameter, E (T) obtained from the simulated delta G(SC), deltal(r)), and net twisting strain of the melted region T (D), is found to be independent of m, hence also of the torsion and bending elastic constants of the melted region. However, E(T) increases linearly with -deltalr), which leads to 1). a small overestimation of E (T) for any given deltal(r)) when E(T) is determined from the observed deltal and deltal (r) by the protocol of Bauer and Benham; and 2). a significant enhancement of the apparent slope, -dE(T)/d(T), obtained via the protocol of Bauer and Benham, relative to the actual slope at fixed delta l(r). After taking these two effects into account, the theoretical and experimental values E(T) and dE(T)/d(T) values agree rather well. For the larger deltal the melted regions are found preferentially in the linker domains between interwound arms, rather than in the apical regions at the ends of interwound arms. PMID- 15111423 TI - Insight into the structural role of carotenoids in the photosystem I: a quantum chemical analysis. AB - The structural stabilization role of carotenoids in the formation of photosynthetic pigment-protein complexes is investigated theoretically. The pi-pi stacking and CH-pi interactions between beta-carotenes and their surrounding chlorophylls (and/or aromatic residues) in Photosystem I (PS1) from the cyanobacterium Synechococcus elongatus were studied by means of the supermolecular approach at the level of the second-order Moller-Plesset perturbation method. PS1 features a core integral antenna system consisting of 22 beta-carotenes intertwined with 90 chlorophyll molecules. The binding environments of all 22 beta-carotenes were systematically analyzed. For 21 out of the 22 cases, one or more chlorophyll molecules exist within van der Waals' contacts of the beta-carotene molecule. The calculated strengths of pi-pi stacking interactions between the conjugated core of beta-carotene and the aromatic tetrapyrrole rings of chlorophyll are substantial, ranging from -3.54 kcal/mol for the perpendicular-positioned BCR4004...CHL1217 pair to -16.01 kcal/mol for the parallel-oriented BCR4007...CHL1122 pair. A strong dependence of the pi-pi stacking interaction energies on the intermolecular configurations of the two interacting pi-planes is observed. The parallel-oriented beta-carotene and chlorophyll pair is energetically much more stable than the perpendicular positioned pair. The larger the extent of pi-pi overlapping, the stronger the interaction strength. In many cases, the beta-ring ends of beta-carotene molecules are found to interact with the tetrapyrrole rings of chlorophyll via CH pi interactions. For the latter interactions, the calculated interaction strengths vary from -7.03 to -11.03 kcal/mol, depending on the intermolecular configuration. This work leads to the conclusion that pi-pi stacking and CH-pi interactions between beta-carotene and their surrounding chlorophylls and aromatic residues play an essential role in binding beta-carotenes in PS1 from S. elongatus. Consequently, the molecular basis of the structural stabilization function of carotenoids in formation of the photosynthetic pigment-protein complexes is established. PMID- 15111424 TI - Role of Arg-72 of pharaonis Phoborhodopsin (sensory rhodopsin II) on its photochemistry. AB - Pharaonis phoborhodopsin (ppR, or pharaonis sensory rhodopsin II, NpsRII) is a sensor for the negative phototaxis of Natronomonas (Natronobacterium) pharaonis. Arginine 72 of ppR corresponds to Arg-82 of bacteriorhodopsin, which is a highly conserved residue among microbial rhodopsins. Using various Arg-72 ppR mutants, we obtained the following results: 1). Arg-72(ppR) together possibly with Asp-193 influenced the pK(a) of the counterion of the protonated Schiff base. 2). The M rise became approximately four times faster than the wild-type. 3). Illumination causes proton uptake and release, and the pH profiles of the sequence of these two proton movements were different between R72A mutant and the wild-type; it is inferred that Arg-72 connects the proton transfer events occurring at both the Schiff base and an extracellular proton-releasing residue (Asp-193). 4). The M decays of Arg-72 mutants were faster ( approximately 8-27 folds at pH 8 depending on mutants) than the wild-type, implying that the guanidinium prevents the proton transfer from the extracellular space to the deprotonated Schiff base. 5), The proton-pumping activities were decreased for mutants having increased M-decay rates, but the extent of the decrease was smaller than expected. The role of Arg 72 of ppR on the photochemistry was discussed. PMID- 15111425 TI - Electrochromic shift of chlorophyll absorption in photosystem I from Synechocystis sp. PCC 6803: a probe of optical and dielectric properties around the secondary electron acceptor. AB - Nanosecond absorption dynamics at approximately 685 nm after excitation of photosystem I (PS I) from Synechocystis sp. PCC 6803 is consistent with electrochromic shift of absorption bands of the Chl a pigments in the vicinity of the secondary electron acceptor A(1). Based on experimental optical data and structure-based simulations, the effective local dielectric constant has been estimated to be between 3 and 20, which suggests that electron transfer in PS I is accompanied by considerable protein relaxation. Similar effective dielectric constant values have been previously observed for the bacterial photosynthetic reaction center and indicate that protein reorganization leading to effective charge screening may be a necessary structural property of proteins that facilitate the charge transfer function. The data presented here also argue against attributing redmost absorption in PS I to closely spaced antenna chlorophylls (Chls) A38 and A39, and suggest that optical transitions of these Chls, along with that of connecting chlorophyll (A40) lie in the range 680-695 nm. PMID- 15111426 TI - Conformation and dynamics of the [3-(13)C]Ala, [1-(13)C]Val-labeled truncated pharaonis transducer, pHtrII(1-159), as revealed by site-directed (13)C solid state NMR: changes due to association with phoborhodopsin (sensory rhodopsin II). AB - We have recorded (13)C NMR spectra of the [3-(13)C]Ala, [1-(13)C]Val-labeled pharaonis transducer pHtrII(1-159) in the presence and absence of phoborhodopsin (ppR or sensory rhodopsin II) in egg phosphatidylcholine or dimyristoylphosphatidylcholine bilayers by means of site-directed (amino acid specific) solid-state NMR. Two kinds of (13)C NMR signals of [3-(13)C]Ala-pHtrII complexed with ppR were clearly seen with dipolar decoupled magic angle spinning (DD-MAS) NMR. One of these resonances was at the peak position of the low-field alpha-helical peaks (alpha(II)-helix) and is identified with cytoplasmic alpha helices protruding from the bilayers; the other was the high-field alpha-helical peak (alpha(I)-helix) and is identified with the transmembrane alpha-helices. The first peaks, however, were almost completely suppressed by cross-polarization magic angle spinning (CP-MAS) regardless of the presence or absence of ppR or by DD-MAS NMR in the absence of ppR. This is caused by an increased fluctuation frequency of the cytoplasmic alpha-helix from 10(5) Hz in the uncomplexed states to >10(6) Hz in the complexed states, leading to the appearance of peaks that were suppressed because of the interference of the fluctuation frequency with the frequency of proton decoupling (10(5) Hz), as viewed from the (13)C NMR spectra of [3-(13)C]Ala-labeled pHtrII. Consistent with this view, the (13)C DD-MAS NMR signals of the cytoplasmic alpha-helices of the complexed [3-(13)C]Ala-pHtrII in the dimyristoylphosphatidylcholine (DMPC) bilayer were partially suppressed at 0 degrees C due to a decreased fluctuation frequency at the low temperature. In contrast, examination of the (13)C CP-MAS spectra of [1-(13)C]Val-labeled complexed pHtrII showed that the (13)C NMR signals of the transmembrane alpha helix were substantially suppressed. These spectral changes are again interpreted in terms of the increased fluctuation frequency of the transmembrane alpha helices from 10(3) Hz of the uncomplexed states to 10(4) Hz of the complexed states. These findings substantiate the view that the transducers alone are in an aggregated or clustered state but the ppR-pHtrII complex is not aggregated. We show that (13)C NMR is a very useful tool for achieving a better understanding of membrane proteins which will serve to clarify the molecular mechanism of signal transduction in this system. PMID- 15111427 TI - The role of cholesterol in the activity of pneumolysin, a bacterial protein toxin. AB - The mechanism via which pneumolysin (PLY), a toxin and major virulence factor of the bacterium Streptococcus pneumoniae, binds to its putative receptor, cholesterol, is still poorly understood. We present results from a series of biophysical studies that shed light on the interaction of PLY with cholesterol in solution and in lipid bilayers. PLY lyses cells whose walls contain cholesterol. Using standard hemolytic assays we have demonstrated that the hemolytic activity of PLY is inhibited by cholesterol, partially by ergosterol but not by lanosterol and that the functional stoichiometry of the cholesterol-PLY complex is 1:1. Tryptophan (Trp) fluorescence data recorded during PLY-cholesterol titration studies confirm this ratio, reveal a significant blue shift in the Trp fluorescence peak with increasing cholesterol concentrations indicative of increasing nonpolarity in the Trp environment, consistent with cholesterol binding by the tryptophans, and provide a measure of the affinity of cholesterol binding: K(d) = 400 +/- 100 nM. Finally, we have performed specular neutron reflectivity studies to observe the effect of PLY upon lipid bilayer structure. PMID- 15111428 TI - The influence of solvent composition on global dynamics of human butyrylcholinesterase powders: a neutron-scattering study. AB - A major result of incoherent elastic neutron-scattering experiments on protein powders is the strong dependence of the intramolecular dynamics on the sample environment. We performed a series of incoherent elastic neutron-scattering experiments on lyophilized human butyrylcholinesterase (HuBChE) powders under different conditions (solvent composition and hydration degree) in the temperature range from 20 to 285 K to elucidate the effect of the environment on the enzyme atomic mean-square displacements. Comparing D(2)O- with H(2)O-hydrated samples, we were able to investigate protein as well as hydration water molecular dynamics. HuBChE lyophilized from three distinct buffers showed completely different atomic mean-square displacements at temperatures above approximately 200 K: a salt-free sample and a sample containing Tris-HCl showed identical small amplitude motions. A third sample, containing sodium phosphate, displayed highly reduced mean-square displacements at ambient temperature with respect to the other two samples. Below 200 K, all samples displayed similar mean-square displacements. We draw the conclusion that the reduction of intramolecular protein mean-square displacements on an Angstrom-nanosecond scale by the solvent depends not only on the presence of salt ions but also on their type. PMID- 15111429 TI - Structure and solvation of melittin in 1,1,1,3,3,3-hexafluoro-2-propanol/water. AB - Fluorinated alcohols can induce peptides and proteins to take up helical conformations. Nuclear Overhauser effect (NOE) spectroscopy experiments and analysis of C(alpha)H proton chemical shifts show that the conformation of melittin in 35% hexafluoro-2-propanol/water is alpha-helical from residues Ile-2 to Val-8 and from Leu-13 to Gln-25. As has been found in other solvent systems, the two helical regions are not colinear; the interhelix angle (73 +/- 15 degrees ) in 35% 1,1,1,3,3,3-hexafluoro-2-propanol/water is smaller than the angle found in other fluoroalcohol-water mixtures or in the crystal. Intermolecular (1)H(19)F and (1)H(1)H nuclear Overhauser effects were used to explore interaction of solvent components with melittin dissolved in this solvent mixture. The NOEs observed indicate that fluoroalcohol and water molecules are both tightly bound to the peptide in the vicinity of the interhelix bend. For the remainder of the molecule, solute-solvent NOEs are consistent with preferential solvation of the peptide by the fluoroalcohol component of the solvent mixture. PMID- 15111430 TI - Temperature derivative fluorescence spectroscopy as a tool to study dynamical changes in protein crystals. AB - Motions through the energy landscape of proteins lead to biological function. At temperatures below a dynamical transition (150-250 K), some of these motions are arrested and the activity of some proteins ceases. Here, we introduce the technique of temperature-derivative fluorescence microspectrophotometry to investigate the dynamical behavior of single protein crystals. The observation of glass transitions in thin films of water/glycerol mixtures allowed us to demonstrate the potential of the technique. Then, protein crystals were investigated, after soaking the samples in a small amount of fluorescein. If the fluorophore resides within the crystal channels, temperature-dependent changes in solvent dynamics can be monitored. Alternatively, if the fluorophore binds to the protein, local dynamical transitions within the biomolecule can be probed directly. A clear dynamical transition was observed at 175 K in the active site of crystalline human butyrylcholinesterase. The results suggest that the dynamics of crystalline proteins is strongly dependent on solvent composition and confinement in the crystal channels. Beyond applications in the field of kinetic crystallography, the highly sensitive temperature-derivative fluorescence microspectrophotometry technique opens the way to many studies on the dynamics of biological nanosamples. PMID- 15111431 TI - Force spectroscopy of collagen fibers to investigate their mechanical properties and structural organization. AB - Tendons are composed of collagen and other molecules in a highly organized hierarchical assembly, leading to extraordinary mechanical properties. To probe the cross-links on the lower level of organization, we used a cantilever to pull substructures out of the assembly. Advanced force probe technology, using small cantilevers (length <20 microm), improved the force resolution into the sub-10 pN range. In the force versus extension curves, we found an exponential increase in force and two different periodic rupture events, one with strong bonds (jumps in force of several hundred pN) with a periodicity of 78 nm and one with weak bonds (jumps in force of <7 pN) with a periodicity of 22 nm. We demonstrate a good correlation between the measured mechanical behavior of collagen fibers and their appearance in the micrographs taken with the atomic force microscope. PMID- 15111432 TI - Mechanism of accelerated assembly of beta-amyloid filaments into fibrils by KLVFFK(6). AB - Extracellular senile plaques are a central pathological feature of Alzheimer's disease. At the core of these plaques are fibrillar deposits of beta-amyloid peptide (Abeta). In vitro, Abeta spontaneously assembles into amyloid fibrils of cross-beta sheet structure. Although it was once believed that the fibrils themselves were toxic, more recent data supports the hypothesis that aggregation intermediates, rather than fully formed fibrils, are the most damaging to neuronal tissue. In previously published work, we identified several small peptides that interact with Abeta and increase its aggregation rate while decreasing its toxicity. In this work, we examined in detail the interaction between Abeta and one of these peptides. Using a mathematical model of Abeta aggregation kinetics, we show that the dominant effect of the peptide is to accelerate lateral association of Abeta filaments into fibrils. PMID- 15111433 TI - Theoretical analysis of twist/bend ratio and mechanical moduli of bacterial flagellar hook and filament. AB - Certain motile bacteria employ rotating flagella for propulsion. The relative flexibility of two key components of the flagellum, filament and hook, is partially responsible for the mechanistic workings of this motor. A new computational method, the quantized elastic deformational model, was employed in this article to calculate the dimensionless twist/bend ratio (EI/GJ) of the filament and hook, providing a quantitative means to compare their relative stiffness. Both ratios were much <1.0, an average of 0.0440 for the filament and 0.0512 for the hook, indicating that within each structure bending is favored over twisting. These two ratios, along with previous experimental measurements, allowed us to propose a theoretical Young's modulus (E) between 10(6) and 10(7) dyn/cm(2) for the hook. This value is orders of magnitude smaller than experimentally determined Young's moduli of the filament, hence in agreement with empirical evidence linking compliance in the flagellum mainly to the hook. PMID- 15111434 TI - Substrate-dependent morphology of supramolecular assemblies: fibrillin and type VI collagen microfibrils. AB - Substrate hydrophobicity/hydrophilicity has previously been shown to affect the morphology and biological function of isolated proteins. We have employed atomic force microscopy to investigate substrate dependent morphologies of two biochemically distinct native supramolecular assemblies: fibrillin and type-VI collagen microfibrils. These morphologically heterogeneous microfibrillar systems are found in many vertebrate tissues where they perform structural and cell signaling roles. Fibrillin microfibrils adsorbed to a hydrophilic mica substrate adopted a diffuse morphology. Fibrillin microfibrils adsorbed to mica coated with poly-L-lysine or to borosilicate glass substrates had a more compact morphology and a directional asymmetry to the bead, which was not present on mica alone. Intermediate morphologies were observed along a substrate gradient. The classical double-beaded appearance of type-VI collagen microfibrils was evident on mica coated with poly-L-lysine and on glass. On hydrophilic mica, morphology was severely disrupted and there was a major conformational reorganization along the whole collagen microfibril repeat. These observations of substrate dependent conformation have important implications for the interpretation of data from in vitro protein interaction assays and cellular signaling studies. Furthermore, conformational changes may be induced by local charge environments in vivo, revealing or hiding binding sites. PMID- 15111435 TI - Multiphoton-excited serotonin photochemistry. AB - We report photochemical and photophysical studies of a multiphoton-excited reaction of serotonin that previously has been shown to generate a photoproduct capable of emitting broadly in the visible spectral region. The current studies demonstrate that absorption of near-infrared light by an intermediate state prepared via three-photon absorption enhances the photoproduct formation yield, with the largest action cross sections ( approximately 10(-19) cm(2)) observed at the short-wavelength limit of the titanium:sapphire excitation source. The intermediate state is shown to persist for at least tens of nanoseconds and likely to be different from a previously reported oxygen-sensitive intermediate. In addition, the two-photon fluorescence action spectrum for the fluorescent photoproduct was determined and found to have a maximum at approximately 780 nm (3.2 eV). A general mechanism for this photochemical process is proposed. PMID- 15111436 TI - Noninvasive auto-photoreduction used as a tool for studying structural changes in heme-copper oxidases by FTIR spectroscopy. AB - We demonstrate an efficient Fourier transform infrared (FTIR) spectroscopic method, termed "auto-photoreduction," that uses anaerobic photo-induced internal electron transfer to monitor reaction-initiated changes of heme-copper oxidases. It can be applied without the use of either expensive electrochemical equipment, or caged compounds, which cause significant background signals. At high irradiation power, carbon monoxide is released from high-spin heme a of cytochrome c oxidase and heme o from cytochrome bo(3). Photochemistry is initiated at wavelengths <355 nm, and the photochemical action spectrum has a maximum of 290 nm for cytochrome bo(3), which is consistent with the possible intermediate involvement of tyrosinate or an activated state of tyrosine. We propose that the final electron donors are proton channel water molecules. In the pH range of 4-9, the noninvasive auto-photoreduction method yields highly reproducible FTIR redox difference spectra within a broad range, resolving a number of vibrational changes outside the amide I region (1600-1640 cm(-1)). Furthermore, it provides details of redox-induced changes in the spectral region between 1600 and 1100 cm(-1). The auto-photoreduction method should be universally applicable to heme proteins. PMID- 15111437 TI - Mean-square displacement relationship in bioprotectant systems by elastic neutron scattering. AB - Neutron intensity elastic scans on trehalose, maltose, and sucrose/H(2)O mixtures as a function of concentration, temperature, and exchanged wave vector are presented. The experimental findings show a crossover in molecular fluctuations between harmonic and anharmonic dynamical regimes. A new operative definition for the degree of fragility of glass-forming systems is furnished by using explicitly the connection between viscosity and mean-square displacement. The procedure is tested for the investigated mixtures and for a set of glass-forming systems. In this frame, the stronger character of trehalose/H(2)O mixture indicates a better attitude in respect to maltose and sucrose/H(2)O mixtures to encapsulate biostructures in a more rigid matrix. PMID- 15111438 TI - Imaging the activity and localization of single voltage-gated Ca(2+) channels by total internal reflection fluorescence microscopy. AB - The patch-clamp technique has enabled functional studies of single ion channels, but suffers limitations including lack of spatial information and inability to independently monitor currents from more than one channel. Here, we describe the use of total internal reflection fluorescence microscopy as an alternative, noninvasive approach to optically monitor the activity and localization of multiple Ca(2+)-permeable channels in the plasma membrane. Images of near membrane Ca(2+) signals were obtained from >100 N-type channels expressed within restricted areas (80 x 80 micro m) of Xenopus oocytes, thereby permitting simultaneous resolution of their gating kinetics, voltage dependence, and localization. Moreover, this technique provided information inaccessible by electrophysiological means, demonstrating that N-type channels are immobile in the membrane, show a patchy distribution, and display diverse gating kinetics even among closely adjacent channels. Total internal reflection fluorescence microscopy holds great promise for single-channel recording of diverse voltage- and ligand-gated Ca(2+)-permeable channels in the membrane of neurons and other isolated or cultured cells, and has potential for high-throughput functional analysis of single channels. PMID- 15111439 TI - Tissue electroporation: quantification and analysis of heterogeneous transport in multicellular environments. AB - Although electroporation is gaining increased attention as a technology to enhance clinical chemotherapy and gene therapy of tissues, direct measurements of electroporation-mediated transport in multicellular environments are lacking. In this study, we used multicellular tumor spheroids of DU145 prostate cancer cells as a model tissue to measure the levels and distribution of molecular uptake in a multicellular environment as a function of electrical and other parameters. These measurements, and subsequent analysis, were used to test the hypothesis that cells in a multicellular environment respond to electroporation in a heterogeneous manner that differs from isolated cells in suspension due to differences in cell state, local solute concentration, and local electric field. In support of the hypothesis, molecular uptake was consistently lower for cells within spheroids than cells in dilute suspension and was spatially heterogeneous, with progressively less uptake observed for cells located deeper within spheroid interiors. Reduced uptake and heterogeneity can be explained quantitatively by accounting for the effects of cell size on transmembrane voltage and cell volume, limited extracellular solute reservoir, heterogeneous field strength due to influence of neighboring cells, and diffusional lag times. PMID- 15111440 TI - Dynamic elastic modulus of porcine articular cartilage determined at two different levels of tissue organization by indentation-type atomic force microscopy. AB - Cartilage stiffness was measured ex vivo at the micrometer and nanometer scales to explore structure-mechanical property relationships at smaller scales than has been done previously. A method was developed to measure the dynamic elastic modulus, |E(*)|, in compression by indentation-type atomic force microscopy (IT AFM). Spherical indenter tips (radius = approximately 2.5 microm) and sharp pyramidal tips (radius = approximately 20 nm) were employed to probe micrometer scale and nanometer-scale response, respectively. |E(*)| values were obtained at 3 Hz from 1024 unloading response curves recorded at a given location on subsurface cartilage from porcine femoral condyles. With the microsphere tips, the average modulus was approximately 2.6 MPa, in agreement with available millimeter-scale data, whereas with the sharp pyramidal tips, it was typically 100-fold lower. In contrast to cartilage, measurements made on agarose gels, a much more molecularly amorphous biomaterial, resulted in the same average modulus for both indentation tips. From results of AFM imaging of cartilage, the micrometer-scale spherical tips resolved no fine structure except some chondrocytes, whereas the nanometer-scale pyramidal tips resolved individual collagen fibers and their 67-nm axial repeat distance. These results suggest that the spherical AFM tip is large enough to measure the aggregate dynamic elastic modulus of cartilage, whereas the sharp AFM tip depicts the elastic properties of its fine structure. Additional measurements of cartilage stiffness following enzyme action revealed that elastase digestion of the collagen moiety lowered the modulus at the micrometer scale. In contrast, digestion of the proteoglycans moiety by cathepsin D had little effect on |E(*)| at the micrometer scale, but yielded a clear stiffening at the nanometer scale. Thus, cartilage compressive stiffness is different at the nanometer scale compared to the overall structural stiffness measured at the micrometer and larger scales because of the fine nanometer-scale structure, and enzyme-induced structural changes can affect this scale-dependent stiffness differently. PMID- 15111441 TI - The effect of alpha-actinin on the length distribution of F-actin. AB - Actin filament length distribution in cells is often regulated to fit specific tasks. In comparison to the well-studied regulation of the average filament length (e.g., using capping proteins), controlling the width of the distribution is less well understood. We utilize two complementary methods to measure the effect of alpha-actinin on the width of the distribution of lengths of F-actin in vitro. Analyzing transmission electron micrographs shows that crosslinking by alpha-actinin reduces the width of the length distribution of F-actin, decreasing the coefficient of variation by two- to threefold. Analysis of fluorescence data from depolymerization assays confirms this observation. We suggest a mechanistic molecular model in which a local (weak) stabilization of crosslinked monomers in the filament is the physical origin of the decrease in the variance of lengths. Although alpha-actinin is known to bind reversibly to F-actin, our model shows that even weak binding can produce this effect, and that in fact it persists throughout a wide range of binding strengths. PMID- 15111442 TI - Specific recognition of macroscopic objects by the cell surface: evidence for a receptor density threshold revealed by micrometric particle binding characteristics. AB - The establishment of specific molecular bonds between a cell and a facing surface is involved in many physiological and technological situations. Using micrometric magnetic particles, we have explored the formation of specific molecular bonds between the cell and surfaces bearing complementary ligands under passive conditions. Streptavidin-coated particles were targeted to the cell surface of a B-cell line through a specific biotinylated antibody against the CD19 receptor. Flow cytometry, optical microscopy, and micropipette experimental techniques have been used. Main findings have been that cell surface receptor density acted like a switch for particle capture with a threshold value found here equal to 1.6 x 10(3) receptor/ microm(2). This led to exclusion from binding of the cells of lowest receptor density. The density threshold was modulated by the length of the binding link and the physics of the cell/particle collision. We suggest that the shear stress is one of the main determinants of the characteristics of binding. We also show that several thousand receptors were involved in the cell particle contact at the end of the binding process, although only eight bonds are required for the initial capture of a particle. A passive binding inhibition process due to link concentration by the initial contact was proposed to account for the small number of particles per cell. PMID- 15111443 TI - Shape memory of human red blood cells. AB - The human red cell can be deformed by external forces but returns to the biconcave resting shape after removal of the forces. If after such shape excursions the rim is always formed by the same part of the membrane, the cell is said to have a memory of its biconcave shape. If the rim can form anywhere on the membrane, the cell would have no shape memory. The shape memory was probed by an experiment called go-and-stop. Locations on the membrane were marked by spontaneously adhering latex spheres. Shape excursions were induced by shear flow. In virtually all red cells, a shape memory was found. After stop of flow and during the return of the latex spheres to the original location, the red cell shape was biconcave. The return occurred by a tank-tread motion of the membrane. The memory could not be eliminated by deforming the red cells in shear flow up to 4 h at room temperature as well as at 37 degrees C. It is suggested that 1). the characteristic time of stress relaxation is >80 min and 2). red cells in vivo also have a shape memory. PMID- 15111444 TI - Cytomechanical properties of papaver pollen tubes are altered after self incompatibility challenge. AB - Self-incompatibility (SI) in Papaver rhoeas triggers a ligand-mediated signal transduction cascade, resulting in the inhibition of incompatible pollen tube growth. Using a cytomechanical approach we have demonstrated that dramatic changes to the mechanical properties of incompatible pollen tubes are stimulated by SI induction. Microindentation revealed that SI resulted in a reduction of cellular stiffness and an increase in cytoplasmic viscosity. Whereas the former cellular response is likely to be the result of a drop in cellular turgor, we hypothesize that the latter is caused by as yet unidentified cross-linking events. F-actin rearrangements, a characteristic phenomenon for SI challenge in Papaver, displayed a spatiotemporal gradient along the pollen tube; this suggests that signal propagation occurs in a basipetal direction. However, unexpectedly, local application of SI inducing S-protein did not reveal any evidence for localized signal perception in the apical or subapical regions of the pollen tube. To our knowledge this represents the first mechanospatial approach to study signal propagation and cellular responses in a well-characterized plant cell system. Our data provide the first evidence for mechanical changes induced in the cytoplasm of a plant cell stimulated by a defined ligand. PMID- 15111445 TI - On the analysis of elastic deformations in hexagonal phases. PMID- 15111447 TI - Normal pressure hydrocephalus (NPH): ischaemia, CSF stagnation or both. PMID- 15111449 TI - Medical research, the media and open access. PMID- 15111452 TI - Drug therapy for Alzheimer's disease. PMID- 15111454 TI - Drug therapy for Alzheimer's disease. PMID- 15111456 TI - Action on Canada's generic drug law. PMID- 15111457 TI - Beyond weapons of mass salvation. PMID- 15111458 TI - Reporting process of randomized controlled trials. PMID- 15111459 TI - New neurosurgeons left jobless: where's the plan? PMID- 15111460 TI - Public health on the installment plan. PMID- 15111461 TI - Haiti emerging from chaos to face health care crisis. PMID- 15111465 TI - A force to contend with: The gender gap closes in Canadian medical schools. PMID- 15111467 TI - Postmenopausal hormone replacement therapy for primary prevention of cardiovascular and cerebrovascular disease. Recommendation statement from the Canadian Task Force on Preventive Health Care. PMID- 15111468 TI - Metabolic syndrome: Waist not want not. PMID- 15111469 TI - Is computed tomographic colonography effective for colorectal cancer screening? PMID- 15111470 TI - A 52-year-old man with cognitive decline, seizure and stroke. PMID- 15111472 TI - Olanzapine (Zyprexa): increased incidence of cerebrovascular events in dementia trials. PMID- 15111473 TI - Do the print media "hype" genetic research? A comparison of newspaper stories and peer-reviewed research papers. AB - BACKGROUND: The public gets most of its information about genetic research from the media. It has been suggested that media representations may involve exaggeration, called "genohype." To examine the accuracy and nature of media coverage of genetic research, we reviewed the reporting of single-gene discoveries and associated technologies in major daily newspapers in Canada, the United States, Great Britain and Australia. METHODS: We used neutral search terms to identify articles about gene discoveries and associated technologies hosted on the Dow Jones Interactive and Canadian NewsDisk databases from January 1995 to June 2001. We compared the contents, claims and conclusions of the scientific journal article with those of the associated newspaper article. Coders subjectively assigned the newspaper articles to 1 of 3 categories: moderately to highly exaggerated claims, slightly exaggerated claims or no exaggerated claims. We used classification tree software to identify the variables that contributed to the assignment of each newspaper article to 1 of the 3 categories: attention structure (positioning in the newspaper and length of the article), authorship, research topic, source of information other than the scientific paper, type and likelihood of risks and benefits, discussion of controversy, valuation tone (positive or negative), framing (e.g., description of research, celebration of progress, report of economic prospects or ethical perspective), technical accuracy (either omissions or errors that changed the description of the methods or interpretation of the results) and use of metaphors. RESULTS: We examined 627 newspaper articles reporting on 111 papers published in 24 scientific and medical journals. Only 11% of the newspaper articles were categorized as having moderately to highly exaggerated claims; the majority were categorized as having no claims (63%) or slightly exaggerated claims (26%). The classification analysis ranked the reporting of risks as the most important variable in determining the categorization of newspaper articles. Only 15% of the newspaper articles and 5% of the scientific journal articles discussed costs or risks, whereas 97% of the newspaper articles and 98% of the scientific journal articles discussed the likelihood of benefits of the research. INTERPRETATION: Our data suggest that the majority of newspaper articles accurately convey the results of and reflect the claims made in scientific journal articles. Our study also highlights an overemphasis on benefits and under-representation of risks in both scientific and newspaper articles. The cause and nature of this trend is uncertain. PMID- 15111474 TI - Gaps in the care of patients admitted to hospital with an exacerbation of chronic obstructive pulmonary disease. AB - BACKGROUND: Patients admitted to hospital because of an exacerbation of chronic obstructive pulmonary disease (COPD) are at high risk of adverse events. We evaluated the association between gaps in care and adverse events during the hospital stay and after discharge. METHODS: We retrospectively reviewed the charts of 105 consecutive patients discharged from hospital between Jan. 1 and Dec. 31, 2001, with a diagnosis of COPD exacerbation. On the basis of published guidelines, prior studies and discussions with colleagues, we defined a care gap as having occurred if any of 9 important inpatient and 7 discharge-related processes of care did not take place correctly. Inpatient adverse events included worsening of condition after admission, transfer to a higher level of care, cardiac arrest and death. Discharge-related adverse events were defined as including readmission to the hospital, revisit to the emergency department or death within 30 days after discharge. RESULTS: Of the 105 patients studied, 88 (84%) had at least 1 inpatient gap in care and 16 (15%) an inpatient adverse event; 2 of the 16 died. Patients who had an inpatient adverse event had more gaps in their care (2.0 v. 1.3 gaps, p = 0.004) and longer stays (16.4 v. 8.6 days, p = 0.007). There were 6 adverse events (frequency 38%) among the 16 patients with 3 or more gaps in their care, 6 adverse events (28%) among the 21 patients with 2 gaps, 1 adverse event (2%) among the 51 patients with 1 gap and 3 adverse events (18%) among the 17 patients with no gaps in their care (p = 0.001 for trend). Of the 103 patients discharged alive, 102 (99%) had at least 1 gap in discharge-related care, but we found no association between these gaps and adverse events within 30 days after discharge. INTERPRETATION: Gaps in the inpatient care of patients with COPD exacerbation were common and were associated with inpatient adverse events. Gaps in discharge-related care were also common but were not associated with postdischarge adverse events. PMID- 15111475 TI - Science reporting to the public: does the message get twisted? PMID- 15111476 TI - Considerations and costs of disclosing study findings to research participants. PMID- 15111477 TI - Physicians, genetics and life insurance. PMID- 15111478 TI - Building a national public health system. PMID- 15111479 TI - Erectile dysfunction: management update. AB - Dramatic advances in the management of erectile dysfunction have occurred over the past decade. Oral therapy with vasoactive agents has emerged as first-line treatment and has transformed both the manner in which the public views erectile dysfunction and the way health care providers deliver care. Whereas an extensive investigation was previously common in the management of erectile dysfunction, recent treatment guidelines promote a more minimalist, goal-oriented approach. In this article, we review the physiology of erection, and the pathophysiology, diagnosis and clinical management of erectile dysfunction. We also present the existing evidence for the efficacy of 3 phosphodiesterase inhibitors, the most widely used class of agents for erectile dysfunction. PMID- 15111484 TI - Genetic studies of the etiology of type 2 diabetes in Pima Indians: hunting for pieces to a complicated puzzle. PMID- 15111485 TI - One week's treatment with the long-acting glucagon-like peptide 1 derivative liraglutide (NN2211) markedly improves 24-h glycemia and alpha- and beta-cell function and reduces endogenous glucose release in patients with type 2 diabetes. AB - Glucagon-like peptide 1 (GLP-1) is potentially a very attractive agent for treating type 2 diabetes. We explored the effect of short-term (1 week) treatment with a GLP-1 derivative, liraglutide (NN2211), on 24-h dynamics in glycemia and circulating free fatty acids, islet cell hormone profiles, and gastric emptying during meals using acetaminophen. Furthermore, fasting endogenous glucose release and gluconeogenesis (3-(3)H-glucose infusion and (2)H(2)O ingestion, respectively) were determined, and aspects of pancreatic islet cell function were elucidated on the subsequent day using homeostasis model assessment and first- and second-phase insulin response during a hyperglycemic clamp (plasma glucose approximately 16 mmol/l), and, finally, on top of hyperglycemia, an arginine stimulation test was performed. For accomplishing this, 13 patients with type 2 diabetes were examined in a double-blind, placebo-controlled crossover design. Liraglutide (6 micro g/kg) was administered subcutaneously once daily. Liraglutide significantly reduced the 24-h area under the curve for glucose (P = 0.01) and glucagon (P = 0.04), whereas the area under the curve for circulating free fatty acids was unaltered. Twenty-four-hour insulin secretion rates as assessed by deconvolution of serum C-peptide concentrations were unchanged, indicating a relative increase. Gastric emptying was not influenced at the dose of liraglutide used. Fasting endogenous glucose release was decreased (P = 0.04) as a result of a reduced glycogenolysis (P = 0.01), whereas gluconeogenesis was unaltered. First-phase insulin response and the insulin response to an arginine stimulation test with the presence of hyperglycemia were markedly increased (P < 0.001), whereas the proinsulin/insulin ratio fell (P = 0.001). The disposition index (peak insulin concentration after intravenous bolus of glucose multiplied by insulin sensitivity as assessed by homeostasis model assessment) almost doubled during liraglutide treatment (P < 0.01). Both during hyperglycemia per se and after arginine exposure, the glucagon responses were reduced during liraglutide administration (P < 0.01 and P = 0.01). Thus, 1 week's treatment with a single daily dose of the GLP-1 derivative liraglutide, operating through several different mechanisms including an ameliorated pancreatic islet cell function in individuals with type 2 diabetes, improves glycemic control throughout 24 h of daily living, i.e., prandial and nocturnal periods. This study further emphasizes GLP-1 and its derivatives as a promising novel concept for treatment of type 2 diabetes. PMID- 15111486 TI - Relationship to insulin resistance of the adult treatment panel III diagnostic criteria for identification of the metabolic syndrome. AB - The Adult Treatment Panel III (ATP III) has published criteria for diagnosing the metabolic syndrome, a cluster of closely related abnormalities related to insulin resistance that increase cardiovascular disease risk. The present analysis was performed to evaluate the ability of these criteria to identify insulin-resistant individuals. The population consisted of 443 healthy volunteers, with measurements of BMI, blood pressure, fasting plasma glucose, triglycerides, HDL cholesterol concentrations, and steady-state plasma glucose (SSPG) concentration. Insulin resistance was defined as being in the top tertile of SSPG concentrations. Of the population, 20% satisfied ATP III criteria for the metabolic syndrome. Although insulin resistance and the presence of the metabolic syndrome were significantly associated (P < 0.001), the sensitivity and positive predictive value equaled 46% (69 of 149) and 76% (69 of 91), respectively. Being overweight, with high triglycerides, low HDL cholesterol, or elevated blood pressure, most often resulted in a diagnosis of the metabolic syndrome. Thus, the ATP III criteria do not provide a sensitive approach to identifying insulin resistant individuals. The individual components vary both in terms of their utility in making a diagnosis of the metabolic syndrome and their relationship to insulin resistance, with the obesity and lipid criteria being most useful. PMID- 15111487 TI - Evaluation of insulin sensitivity and beta-cell function indexes obtained from minimal model analysis of a meal tolerance test. AB - Modeling analysis of glucose, insulin, and C-peptide following a meal has been proposed as a means to estimate insulin sensitivity (S(i)) and beta-cell function from a single test. We compared the model-derived meal indexes with analogous indexes obtained from an intravenous glucose tolerance test (IVGTT) and hyperglycemic clamp (HGC) in 17 nondiabetic subjects (14 men, 3 women, aged 50 +/ 2 years [mean +/- SE], BMI 25.0 +/- 0.7 kg/m(2)). S(i) estimated from the meal was correlated with S(i) estimated from the IVGTT and the HGC (r = 0.59 and 0.76, respectively; P < 0.01 for both) but was approximately 2.3 and 1.4 times higher (P < 0.05 for both). The meal-derived estimate of the beta-cell's response to a steady-state change in glucose (static secretion index) was correlated with the HGC second-phase insulin response (r = 0.69; P = 0.002), but the estimated rate of-change component (dynamic secretion index) was not correlated with first-phase insulin release from either the HGC or IVGTT. Indexes of beta-cell function obtained from the meal were significantly higher than those obtained from the HGC. In conclusion, insulin sensitivity and beta-cell indexes derived from a meal are not analogous to those from the clamp or IVGTT. Further work is needed before these indexes can be routinely used in clinical and epidemiological studies. PMID- 15111488 TI - Exercise and myocyte enhancer factor 2 regulation in human skeletal muscle. AB - Overexpression of GLUT4 in skeletal muscle enhances whole-body insulin action. Exercise increases GLUT4 gene and protein expression, and a binding site for the myocyte enhancer factor 2 (MEF-2) is required on the GLUT4 promoter for this response. However, the molecular mechanisms involved remain elusive. In various cell systems, MEF-2 regulation is a balance between transcriptional repression by histone deacetylases (HDACs) and transcriptional activation by the nuclear factor of activated T-cells (NFAT), peroxisome proliferator-activated receptor-gamma coactivator 1 (PGC-1), and the p38 mitogen-activated protein kinase. The purpose of this study was to determine if these same mechanisms regulate MEF-2 in contracting human skeletal muscle. Seven subjects performed 60 min of cycling at approximately 70% of VO2(peak). After exercise, HDAC5 was dissociated from MEF-2 and exported from the nucleus, whereas nuclear PGC-1 was associated with MEF-2. Exercise increased total and nuclear p38 phosphorylation and association with MEF 2, without changes in total or nuclear p38 protein abundance. This result was associated with p38 sequence-specific phosphorylation of MEF-2 and an increase in GLUT4 mRNA. Finally, we found no role for NFAT in MEF-2 regulation. From these data, it appears that HDAC5, PGC-1, and p38 regulate MEF-2 and could be potential targets for modulating GLUT4 expression. PMID- 15111489 TI - Relationship between insulin sensitivity and sphingomyelin signaling pathway in human skeletal muscle. AB - In vitro studies revealed that insulin resistance might be associated with the intracellular formation of ceramide, the second messenger in the sphingomyelin signaling pathway. The aim of the present study was to examine the content and composition of fatty acids in ceramide and sphingomyelin in human muscle and to evaluate their relationships with insulin sensitivity. The study was conducted on 27 male subjects with normal glucose tolerance. Euglycemic-hyperinsulinemic clamps and biopsies of vastus lateralis muscle were performed. In 10 subjects, additional biopsies were taken after a 4-h clamp and after a clamp with concurrent Intralipid/heparin infusion. We identified 13 ceramides and sphingomyelins according to fatty acid residues. Insulin sensitivity was related to total ceramide content (r = -0.49, P = 0.01) and to ceramide consisting of palmitic (r = -0.48, P = 0.011), palmitoleic (r = -0.45, P = 0.019), mirystic (r = -0.42, P = 0.028), and nervonic acid (r = -0.39, P = 0.047). Hyperinsulinemia did not affect estimated muscle parameters. Intralipid/heparin infusion resulted in a 24.73% decrease in insulin sensitivity (P = 0.007) and a 47.81% increase in ceramide content (P = 0.005). These changes were significantly related to each other (r = -0.64, P = 0.046). A relationship with the decrease in insulin sensitivity was also observed for ceramides consisting of palmitic (r = -0.68, P = 0.03) and linoleic (r = -0.66, P = 0.038) acid. Our data indicate that the sphingomyelin signaling pathway in muscle might be an important factor determining the development of insulin resistance in humans. PMID- 15111490 TI - Bifunctional properties of peroxisome proliferator-activated receptor gamma1 in KDR gene regulation mediated via interaction with both Sp1 and Sp3. AB - Vascular endothelial growth factor receptor 2 (KDR) plays a critical role in mediating a variety of vasculogenic and angiogenic processes, including diabetic retinopathy. We previously demonstrated that the promoter activity of the KDR gene in retinal capillary endothelial cells (RCECs) was regulated in part by the relative concentration of positive/negative transcription factors Sp1/Sp3. We also reported that the peroxisome proliferator-activated receptor (PPAR)gamma ligand could inhibit intraocular angiogenesis. In the present study, the role of PPARgamma1 in KDR gene regulation in RCECs was examined. PPARgamma1 protein physically interacted with both Sp1 and Sp3. Transactivation and electrophoretic mobility shift assays clearly demonstrated novel findings that PPARgamma1 increased KDR promoter activity by enhancing the interaction between Sp1, but not Sp3, and KDR promoter region without its ligand in RCECs. The ligand-binding site but not the DNA binding site of PPARgamma1 enhanced the interaction between Sp1 and KDR promoter region. Conversely, PPARgamma1 ligand 15-deoxy Delta (12,14) prostaglandin J2 dose-dependently suppressed the binding of KDR promoter region with both Sp1 and Sp3, resulting an inhibition of KDR gene expression. In conclusion, PPARgamma1 has bifunctional properties in the regulation of KDR gene expression mediated via interaction with both Sp1 and Sp3. PMID- 15111491 TI - Third ventricular alloxan reversibly impairs glucose counterregulatory responses. AB - Glucokinase (GK) is hypothesized to be the critical glucosensor of pancreatic beta-cells and hypothalamic glucosensing neurons. To understand the role of GK in glucoprivic counterregulatory responses, we injected alloxan, a GK inhibitor and toxin, into the third ventricle (3v) to target nearby GK-expressing neurons. Four and 6 days after 3v, but not 4v, alloxan injection, alloxan-treated rats ate only 30% and their blood glucose area under the curve was only 28% of saline controls' after systemic 2-deoxy-D-glucose. In addition, their hyperglycemic response to hindbrain glucoprivation induced with 5-thio-glucose was impaired, whereas fasting blood glucose levels and food intake after an overnight fast were elevated. These impaired responses were associated with the destruction of 3v tanycytes, reduced glial fibrillary acidic protein-immunoreactivity surrounding the 3v, neuronal swelling, and decreased arcuate nucleus neuropeptide Y (NPY) mRNA. Nevertheless, hypothalamic GK mRNA was significantly elevated. Two weeks after alloxan injection, 3v tanycyte destruction was reversed along with restoration of feeding and hyperglycemic responses to both systemic and hindbrain glucoprivation. At this time there were significant decreases in GK, NPY, and proopiomelanocortin mRNA. Thus, neural substrates near and around the 3v affected by alloxan may be critically involved in the expression of these glucoprivic responses. PMID- 15111492 TI - Effects of diabetes and insulin on the expression of galanin-like peptide in the hypothalamus of the rat. AB - Galanin-like peptide (GALP) is produced in a small population of neurons in the arcuate nucleus of the hypothalamus, and leptin stimulates the hypothalamic expression of GALP mRNA. Because insulin and leptin share common signaling pathways in the brain, we reasoned that GALP neurons might also be responsive to changes in circulating concentrations of insulin. To test this hypothesis, we first studied the effect of insulin deficiency on the expression of GALP by comparing levels of GALP mRNA between normal and diabetic animals. Streptozotocin induced diabetes was associated with a significant reduction in the expression of GALP mRNA, which was reversed by treatment with either insulin or leptin. Second, we examined the effect of insulin administered directly into the brain on the expression of GALP mRNA in fasted rats. Hypothalamic levels of GALP mRNA were lower in animals after a 48-h fast, and central treatment with insulin reversed this effect. These results suggest that GALP neurons are direct targets for regulation by insulin and implicate these cells for a role in the metabolic and behavioral sequelae of type 1 diabetes. PMID- 15111493 TI - Adipose tissue expression of the lipid droplet-associating proteins S3-12 and perilipin is controlled by peroxisome proliferator-activated receptor-gamma. AB - In a systematic search for peroxisome proliferator-activated receptor-gamma (PPAR gamma) target genes, we identified S3-12 and perilipin as novel direct PPAR-gamma target genes. Together with adipophilin and tail-interacting protein of 47 kDa, these genes are lipid droplet-associating proteins with distinct expression pattern but overlapping expression in adipose tissue. The expression of S3-12 and perilipin is tightly correlated to the expression and activation of PPAR-gamma in adipocytes, and promoter characterization revealed that the S3-12 and the perilipin promoters contain three and one evolutionarily conserved PPAR response elements, respectively. We furthermore demonstrate that the expression of S3-12 and perilipin is reduced in obese compared with lean Zucker rats, whereas the expression of adipophilin is increased. Others have shown that perilipin is an essential factor in the hormonal regulation of lipolysis of stored triglycerides within adipose tissue. The direct regulation of perilipin and S3-12 by PPAR-gamma therefore is likely to be an important mediator of the in vivo effects of prolonged treatment with PPAR-gamma activators: insulin sensitization, fatty acid trapping in adipose tissue, reduced basal adipose lipolysis, and weight gain. PMID- 15111494 TI - Triglycerides induce leptin resistance at the blood-brain barrier. AB - Obesity is associated with leptin resistance as evidenced by hyperleptinemia. Resistance arises from impaired leptin transport across the blood-brain barrier (BBB), defects in leptin receptor signaling, and blockades in downstream neuronal circuitries. The mediator of this resistance is unknown. Here, we show that milk, for which fats are 98% triglycerides, immediately inhibited leptin transport as assessed with in vivo, in vitro, and in situ models of the BBB. Fat-free milk and intralipid, a source of vegetable triglycerides, were without effect. Both starvation and diet-induced obesity elevated triglycerides and decreased the transport of leptin across the BBB, whereas short-term fasting decreased triglycerides and increased transport. Three of four triglycerides tested intravenously inhibited transport of leptin across the BBB, but their free fatty acid constituents were without effect. Treatment with gemfibrozil, a drug that specifically reduces triglyceride levels, reversed both hypertriglyceridemia and impaired leptin transport. We conclude that triglycerides are an important cause of leptin resistance as mediated by impaired transport across the BBB and suggest that triglyceride-mediated leptin resistance may have evolved as an anti anorectic mechanism during starvation. Decreasing triglycerides may potentiate the anorectic effect of leptin by enhancing leptin transport across the BBB. PMID- 15111495 TI - Role of caveolin-1 in the modulation of lipolysis and lipid droplet formation. AB - Recently, it was shown that caveolin-1 can be redirected from the cell surface to intracellular lipid droplets in a variety of cell types. Here, we directly address the role of caveolin-1 in lipid droplet formation and breakdown, showing that caveolin-1 null mice exhibit markedly attenuated lipolytic activity. Mechanistically, although the activity of protein kinase A (PKA) was greatly increased in caveolin-1 null adipocytes, the phosphorylation of perilipin was dramatically reduced, indicating that caveolin-1 may facilitate the PKA-mediated phosphorylation of perilipin. In support of this hypothesis, coimmunoprecipitation experiments revealed that treatment with a beta(3) adrenergic receptor agonist resulted in ligand-induced complex formation between perilipin, caveolin-1, and the catalytic subunit of PKA in wild-type but not in caveolin-1 null fat pads. We also show that caveolin-1 expression is important for efficient lipid droplet formation because caveolin-1 null embryonic fibroblasts stably transfected with perilipin accumulated approximately 4.5-fold less lipid than perilipin-transfected wild-type cells. Finally, high-pressure freeze-substitution electron microscopy of adipose tissue revealed dramatic perturbations in the architecture of the "lipid droplet cortex" (the interface between the lipid droplet surface and the cytoplasm) in caveolin-1 null perigonadal adipocytes. Taken together, our data provide the first molecular genetic evidence that caveolin-1 plays a critical functional and structural role in the modulation of both lipid droplet biogenesis and metabolism in vivo. PMID- 15111496 TI - Effects of dietary monosaccharides on sympathetic nervous system activity in adipose tissues of male rats. AB - Dietary carbohydrate activates the sympathetic nervous system (SNS). As the mechanisms underlying this response are not fully characterized, studies were undertaken to compare SNS responses to ingestion of glucose, fructose, and galactose. SNS activity was examined using techniques of [(3)H]norepinephrine ([(3)H]NE) turnover in brown and white fat. In addition, gene expression for several sympathetically related proteins was also analyzed in these tissues. [(3)H]NE turnover in interscapular brown adipose tissue (IBAT) and retroperitoneal fat increased in response to glucose and fructose in the diet, whereas [(3)H]NE turnover in epididymal fat did not respond to either monosaccharide. Galactose feeding, by contrast, decreased [(3)H]NE turnover in IBAT, but increased it in epididymal, though not retroperitoneal, fat. Expression of GLUT4 was more abundant in IBAT and retroperitoneal fat from glucose- and fructose-fed animals than from diet- or galactose-fed rats. Chemical sympathectomy abolished the GLUT4 response in retroperitoneal fat, but was without effect on GLUT4 in epididymal fat. These studies are consistent with activation of a neural pathway by oral glucose or fructose, leading to SNS activation in IBAT and retroperitoneal fat and enhanced GLUT4 expression. PMID- 15111497 TI - High serum resistin is associated with an increase in adiposity but not a worsening of insulin resistance in Pima Indians. AB - Resistin is an adipokine with putative prodiabetogenic properties. Like other hormones secreted by adipose tissue, resistin is being investigated as a possible etiologic link between excessive adiposity and insulin resistance. Although there is growing evidence that circulating levels of this adipokine are proportional to the degree of adiposity, an effect on insulin resistance in humans remains unproven. To evaluate the relations among resistin, obesity, and insulin resistance, we measured fasting serum resistin levels in 113 nondiabetic (75-g oral glucose tolerance test) Pima Indians (ages 29 +/- 7 years, body fat 31 +/- 8%, resistin 3.7 +/- 1.1 ng/ml [means +/- SD]), who were characterized for body composition (assessed by hydrodensitometry or dual-energy X-ray absorptiometry), whole-body insulin sensitivity (M; assessed by hyperinsulinemic clamp), basal hepatic glucose output (BHGO) and hepatic glucose output during low-dosage insulin infusion of a hyperinsulinemic clamp (HGO; a measure of hepatic insulin resistance), and acute insulin secretory response (AIR; assessed by 25-g intravenous glucose tolerance test). Follow-up measurements of M, BHGO, HGO, and AIR were available for 34 subjects who had normal glucose tolerance at baseline and remained nondiabetic at follow-up. The average time to follow-up was 4.5 +/- 2.7 years. In cross-sectional analyses, serum resistin levels were positively associated with percent body fat (r = 0.37, P = 0.0001) and 2-h glucose (r = 0.19, P = 0.04), respectively. Serum resistin levels were not associated with fasting glucose and insulin levels, M, BHGO, HGO, or AIR (r = 0.17, 0.12, -0.13, 0.06, -0.03, and -0.04, respectively; all P > 0.05). After adjusting for percent body fat, there was no association between serum resistin levels and 2-h glucose (r = 0.06, P = 0.6). In prospective analyses, high serum resistin levels at baseline were not associated with a decline in M (r = -0.1, P > 0.5). Resistin levels were, however, associated with increases in percent body fat, fasting plasma insulin, and HGO (r = 0.34, 0.36, and 0.37; all P < 0.05) after adjusting for sex, age, and time to follow-up. After additional adjustment for the change in percent body fat, there was no association between baseline serum resistin levels and changes in plasma insulin or HGO (r = 0.26 and 0.23; both P > 0.1). We conclude that in Pima Indians, like other human populations, circulating resistin levels are proportional to the degree of adiposity, but not the degree of insulin resistance. We unexpectedly found that high serum resistin levels do predict future increases in percent body fat. Our data suggest that resistin promotes obesity but not obesity-associated insulin resistance in humans. PMID- 15111498 TI - From blood monocytes to adipose tissue-resident macrophages: induction of diapedesis by human mature adipocytes. AB - Obesity has been suggested to be a low-grade systemic inflammatory state, therefore we studied the interaction between human adipocytes and monocytes via adipose tissue (AT)-derived capillary endothelium. Cells composing the stroma vascular fraction (SVF) of human ATs were characterized by fluorescence-activated cell sorter (FACS) analysis and two cell subsets (resident macrophages and endothelial cells [ECs]) were isolated using antibody-coupled microbeads. Media conditioned by mature adipocytes maintained in fibrin gels were applied to AT derived ECs. Thereafter, the expression of endothelial adhesion molecules was analyzed as well as the adhesion and transmigration of human monocytes. FACS analysis showed that 11% of the SVF is composed of CD14(+)/CD31(+) cells, characterized as resident macrophages. A positive correlation was found between the BMI and the percentage of resident macrophages, suggesting that fat tissue growth is associated with a recruitment of blood monocytes. Incubation of AT derived ECs with adipocyte-conditioned medium resulted in the upregulation of EC adhesion molecules and the increased chemotaxis of blood monocytes, an effect mimicked by recombinant human leptin. These results indicate that adipokines, such as leptin, activate ECs, leading to an enhanced diapedesis of blood monocytes, and suggesting that fat mass growth might be linked to inflammatory processes. PMID- 15111499 TI - Islet cell autoimmunity in a triethnic adult population of the Third National Health and Nutrition Examination Survey. AB - Markers of humoral islet cell autoimmunity, such as autoantibodies (AAs) against the 65-kDa isoform of GAD (GAD65), serve as determinants of risk for autoimmune diabetes. Despite the high prevalence of diabetes in U.S. racial and ethnic minority adult populations, little is known concerning the prevalence of GAD65 AA in these groups. We estimated the prevalence of GAD65 AA in 1,064 diabetic and 1,036 nondiabetic participants who were 40-90 years of age from the Third National Health and Nutrition Examination Survey (NHANES III), which provides a representative ethnic sample of the U.S. diabetic population. The prevalence of GAD65 AA was higher in diabetic participants compared with nondiabetic participants in non-Hispanic whites (n = 920; 6.3% vs. 2.0%; P = 0.001) and non Hispanic blacks (n = 534; 3.7% vs. 1.3%; P = 0.08) but not in Mexican Americans (n = 646; 1.2% vs. 2.6%; P = 0.18). Among diabetic non-Hispanic whites and non Hispanic blacks, being GAD65 AA positive was associated with lower BMI and C peptide (P < 0.05). These results may reflect the outcome of an autoimmune process leading to beta-cell destruction/dysfunction in non-Hispanic white and non-Hispanic black adult diabetic patients as it occurs in a similar manner in type 1 diabetes. Among diabetic Mexican Americans, the lower prevalence of GAD65 AA suggests a lower frequency of autoimmune-related diabetes. PMID- 15111500 TI - Interleukin-4 but not interleukin-10 protects against spontaneous and recurrent type 1 diabetes by activated CD1d-restricted invariant natural killer T-cells. AB - In nonobese diabetic (NOD) mice, a deficiency in the number and function of invariant natural killer T-cells (iNKT cells) contributes to the onset of type 1 diabetes. The activation of CD1d-restricted iNKT cells by alpha galactosylceramide (alpha-GalCer) corrects these deficiencies and protects against spontaneous and recurrent type 1 diabetes. Although interleukin (IL)-4 and IL-10 have been implicated in alpha-GalCer-induced protection from type 1 diabetes, a precise role for these cytokines in iNKT cell regulation of susceptibility to type 1 diabetes has not been identified. Here we use NOD.IL-4( /-) and NOD.IL-10(-/-) knockout mice to further evaluate the roles of IL-4 and IL 10 in alpha-GalCer-induced protection from type 1 diabetes. We found that IL-4 but not IL-10 expression mediates protection against spontaneous type 1 diabetes, recurrent type 1 diabetes, and prolonged syngeneic islet graft function. Increased transforming growth factor-beta gene expression in pancreatic lymph nodes may be involved in alpha-GalCer-mediated protection in NOD.IL-10(-/-) knockout mice. Unlike the requirement of IL-7 and IL-15 to maintain iNKT cell homeostasis, IL-4 and IL-10 are not required for alpha-GalCer-induced iNKT cell expansion and/or survival. Our data identify an important role for IL-4 in the protection against type 1 diabetes by activated iNKT cells, and these findings have important implications for cytokine-based therapy of type 1 diabetes and islet transplantation. PMID- 15111501 TI - Prevalence of hepatic steatosis after islet transplantation and its relation to graft function. AB - Islet allotransplantation can provide insulin independence in selected individuals with type 1 diabetes. The long-term effects of these transplants on the liver are unknown. Recently, two cases of periportal steatosis after islet transplantation have been described. In this study, we performed ultrasound and magnetic resonance imaging (MRI) in 30 C-peptide-positive islet transplant recipients to detect steatosis and to explore the association of the radiological findings with clinical and metabolic factors. Steatosis was observed on MRI in six (20%) subjects. Histological findings of hepatic steatosis concurred with the imaging findings. Steatosis completely resolved in one subject whose graft failed. More subjects with steatosis required supplementary exogenous insulin than not (67 vs. 21%; P < 0.05). The clinical features of subjects with and without steatosis were otherwise similar, although C-peptide levels were higher in insulin-independent subjects with steatosis (0.98 +/- 0.12 vs. 0.70 +/- 0.18 nmol/l; P = 0.05), despite similar blood glucose levels. Serum triglycerides and the use of exogenous insulin were associated with increased odds of steatosis in a logistic regression model (chi(2) [degrees freedom] = 13.6 [2]); P = 0.001). MRI-detected steatosis is a common finding; the steatosis appears to be due to a paracrine action of insulin secreted from intrahepatic islets. Hepatic steatosis may be associated with insulin resistance or graft dysfunction. PMID- 15111502 TI - Intraislet endothelial cells contribute to revascularization of transplanted pancreatic islets. AB - Pancreatic islet transplantation is an emerging therapy for type 1 diabetes. To survive and function, transplanted islets must revascularize because islet isolation severs arterial and venous connections; the current paradigm is that islet revascularization originates from the transplant recipient. Because isolated islets retain intraislet endothelial cells, we determined whether these endothelial cells contribute to the revascularization using a murine model with tagged endothelial cells (lacZ knock-in to Flk-1/VEGFR2 gene) and using transplanted human islets. At 3-5 weeks after transplantation beneath the renal capsule, we found that islets were revascularized and that the transplant recipient vasculature indeed contributed to the revascularization process. Using the lacZ-tagged endothelial cell model, we found that intraislet endothelial cells not only survived after transplantation but became a functional part of revascularized islet graft. A similar contribution of intraislet endothelial cells was also seen with human islets transplanted into an immunodeficient mouse model. In the murine model, individual blood vessels within the islet graft consisted of donor or recipient endothelial cells or were a chimera of donor and recipient endothelial cells, indicating that both sources of endothelial cells contribute to the new vasculature. These observations suggest that interventions to activate, amplify, or sustain intraislet endothelial cells before and after transplantation may facilitate islet revascularization, enhance islet survival, and improve islet transplantation. PMID- 15111503 TI - Double incretin receptor knockout (DIRKO) mice reveal an essential role for the enteroinsular axis in transducing the glucoregulatory actions of DPP-IV inhibitors. AB - Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide 1 (GLP-1) are gut-derived incretins that potentiate glucose clearance following nutrient ingestion. Elimination of incretin receptor action in GIPR(-/-) or GLP 1R(-/-) mice produces only modest impairment in glucose homeostasis, perhaps due to compensatory upregulation of the remaining incretin. We have now studied glucose homeostasis in double incretin receptor knockout (DIRKO) mice. DIRKO mice exhibit normal body weight and fail to exhibit an improved glycemic response after exogenous administration of GIP or the GLP-1R agonist exendin-4. Plasma glucagon and the hypoglycemic response to exogenous insulin were normal in DIRKO mice. Glycemic excursion was abnormally increased and levels of glucose stimulated insulin secretion were decreased following oral but not intraperitoneal glucose challenge in DIRKO compared with GIPR(-/-) or GLP-1R(-/-) mice. Similarly, glucose-stimulated insulin secretion and the response to forskolin were well preserved in perifused DIRKO islets. Although the dipeptidyl peptidase-IV (DPP-IV) inhibitors valine pyrrolidide (Val-Pyr) and SYR106124 lowered glucose and increased plasma insulin in wild-type and single incretin receptor knockout mice, the glucose-lowering actions of DPP-IV inhibitors were eliminated in DIRKO mice. These findings demonstrate that glucose-stimulated insulin secretion is maintained despite complete absence of both incretin receptors, and they delineate a critical role for incretin receptors as essential downstream targets for the acute glucoregulatory actions of DPP-IV inhibitors. PMID- 15111504 TI - Catalase protects cardiomyocyte function in models of type 1 and type 2 diabetes. AB - Many diabetic patients suffer from a cardiomyopathy that cannot be explained by poor coronary perfusion. Reactive oxygen species (ROS) have been proposed to contribute to this cardiomyopathy. Consistent with this we found evidence for induction of the antioxidant genes for catalase in diabetic OVE26 hearts. To determine whether increased antioxidant protection could reduce diabetic cardiomyopathy, we assessed cardiac morphology and contractility, Ca(2+) handling, malondialdehyde (MDA)-modified proteins, and ROS levels in individual cardiomyocytes isolated from control hearts, OVE26 diabetic hearts, and diabetic hearts overexpressing the antioxidant protein catalase. Diabetic hearts showed damaged mitochondria and myofibrils, reduced myocyte contractility, slowed intracellular Ca(2+) decay, and increased MDA-modified proteins compared with control myocytes. Overexpressing catalase preserved normal cardiac morphology, prevented the contractile defects, and reduced MDA protein modification but did not reverse the slowed Ca(2+) decay induced by diabetes. Additionally, high glucose promoted significantly increased generation of ROS in diabetic cardiomyocytes. Chronic overexpression of catalase or acute in vitro treatment with rotenone, an inhibitor of mitochondrial complex I, or thenoyltrifluoroacetone, an inhibitor of mitochondrial complex II, eliminated excess ROS production in diabetic cardiomyocytes. The structural damage to diabetic mitochondria and the efficacy of mitochondrial inhibitors in reducing ROS suggest that mitochondria are a source of oxidative damage in diabetic cardiomyocytes. We also found that catalase overexpression protected cardiomyocyte contractility in the agouti model of type 2 diabetes. These data show that both type 1 and type 2 diabetes induce damage at the level of individual myocytes, and that this damage occurs through mechanisms utilizing ROS. PMID- 15111505 TI - Insulin generates free radicals by an NAD(P)H, phosphatidylinositol 3'-kinase dependent mechanism in human skin fibroblasts ex vivo. AB - Oxidative stress may be involved in the development of vascular complications associated with diabetes; however, the molecular mechanism responsible for increased production of free radicals in diabetes remains uncertain. Therefore, we examined whether acute hyperinsulinemia increases the production of free radicals and whether this condition affects proliferative extracellular signal regulated kinase (ERK-1 and -2) signaling in human fibroblasts in vitro. Insulin treatment significantly increased intracellular superoxide anion (O(2)(-)) production, an effect completely abolished by Tiron, a cell-permeable superoxide dismutase (SOD) mimetic and by polyethylene glycol (PEG)-SOD, but not by PEG catalase. Furthermore, insulin-induced O(2)(-) production was attenuated by the NAD(P)H inhibitor apocynin, but not by rotenone or oxypurinol. Inhibition of the phosphatidylinositol 3'-kinase (PI 3'-kinase) pathway with LY294002 blocked insulin-stimulated O(2)(-) production, suggesting a direct involvement of PI 3' kinase in the activation of NAD(P)H oxidase. The insulin-induced free radical production led to membranous translocation of p47phox and markedly enhanced ERK-1 and -2 activation in human fibroblasts. In conclusion, these findings provided direct evidence that elevated insulin levels generate O(2)(-) by an NAD(P)H dependent mechanism that involves the activation of PI 3'-kinase and stimulates ERK-1- and ERK-2-dependent pathways. This effect of insulin may contribute to the pathogenesis and progression of cardiovascular disease in the insulin resistance syndrome. PMID- 15111506 TI - Cerebrovascular dysfunction in Zucker obese rats is mediated by oxidative stress and protein kinase C. AB - Insulin resistance (IR) impairs vascular function in the peripheral and coronary circulations, but its effects on cerebral arteries are virtually unexplored. We examined the vascular responses of the basilar artery (BA) and its side branches through a cranial window in Zucker lean (ZL) and IR Zucker obese (ZO) rats. Nitric oxide (NO) and K+ channel-mediated dilator responses, elicited by acetylcholine, iloprost, cromakalim, and elevated [K+], were greatly diminished in the ZO rats compared with ZL rats. In contrast, sodium nitroprusside induced similar relaxations in the two experimental groups. Expressions of the K+ channel pore-forming subunits were not affected by IR, while endothelial NO synthase was upregulated in the ZO arteries compared with ZL arteries. Protein kinase C (PKC) activity and production of superoxide anion were increased in the cerebral arteries of ZO rats, and pretreatment with superoxide dismutase restored all examined dilator responses. In contrast, application of PKC inhibitors improved only receptor-linked NO-mediated relaxation, but not K+ channel-dependent responses. Thus, IR induces in ZO rats cerebrovascular dysfunction, which is mediated by oxidative stress and partly by PKC activation. The revealed impairment of NO and K+ channel-dependent dilator responses may be responsible for the increased risk of cerebrovascular events and neurodegenerative disorders in IR. PMID- 15111507 TI - Haplotype structure and genotype-phenotype correlations of the sulfonylurea receptor and the islet ATP-sensitive potassium channel gene region. AB - The genes for the sulfonylurea receptor (SUR1; encoded by ABCC8) and its associated islet ATP-sensitive potassium channel (Kir6.2; encoded by KCNJ11) are adjacent to one another on human chromosome 11. Multiple studies have reported association of the E23K variant of Kir6.2 with risk of type 2 diabetes. Whether and how E23K itself-or other variant(s) in either of these two closely linked genes-influences type 2 diabetes remains to be fully determined. To better understand genotype-phenotype correlation at this important candidate gene locus, we 1) characterized haplotype structures across the gene region by typing 77 working, high-frequency markers spanning 207 kb and both genes; 2) performed association studies of E23K and nearby markers in >3,400 patients (type 2 diabetes and control) not previously reported in the literature; and 3) analyzed the resulting data for measures of insulin secretion. These data independently replicate the association of E23K with type 2 diabetes with an odds ratio (OR) in the new data of 1.17 (P = 0.003) as compared with an OR of 1.14 provided by meta analysis of previously published, nonoverlapping data (P = 0.0002). We find that the E23K variant in Kir6.2 demonstrates very strong allelic association with a coding variant (A1369S) in the neighboring SUR1 gene (r(2) > 0.9) across a range of population samples, making it difficult to distinguish which gene and polymorphism in this region are most likely responsible for the reported association. We show that E23K is also associated with decreased insulin secretion in glucose-tolerant control subjects, supporting a mechanism whereby beta-cell dysfunction contributes to the common form of type 2 diabetes. Like peroxisome proliferator-activated receptor gamma, the SUR1/Kir6.2 gene region both contributes to the inherited risk of type 2 diabetes and encodes proteins that are targets for hypoglycemic medications, providing an intriguing link between the underlying mechanism of disease and validated targets for pharmacological treatment. PMID- 15111508 TI - Genome-wide scans reveal quantitative trait Loci on 8p and 13q related to insulin action and glucose metabolism: the San Antonio Family Heart Study. AB - Type 2 diabetes is a complex disease that arises from physiological disruptions of the body's sensitivity to insulin and ability to metabolize glucose. Multipoint linkage analyses for insulin sensitivity phenotypes were conducted in 1,280 Mexican Americans from 41 families who participated in the San Antonio Family Heart Study. A significant linkage signal (logarithm of odds [LOD] = 2.98) affecting corrected insulin response to glucose was detected on chromosome 13q between D13787 and D13S252, in the region where the MODY-4 gene has previously been mapped. Another signal on chromosome 13 was observed at D13S285 (LOD = 1.86), where the insulin receptor substrate 2 gene resides. Significant linkage (LOD = 3.09) for insulin response to glucose was found on chromosome 8 between D8S1130 and D8S1106, near the lipoprotein lipase and macrophage scavenger receptor genes. Multipoint analysis of abdominal skinfold with an LOD of 2.68 showed signals in the same region. There was also suggestive evidence for linkage of quantitative insulin sensitivity check index and fasting glucose to a previously reported location at D9S301 (LOD = 2.19). These results indicate that chromosomal locations on 8p and 13q might harbor genes that affect a variety of insulin- and glucose-related phenotypes that contribute to the observed variations in these important risk factors for diabetes in Mexican Americans. PMID- 15111509 TI - Identification of a locus for maturity-onset diabetes of the young on chromosome 8p23. AB - Maturity-onset diabetes of the young (MODY) is a subtype of diabetes defined by an autosomal dominant inheritance and a young onset. Six MODY genes have been discovered to date. To identify additional MODY loci, we conducted a genome scan in 21 extended U.S. families (15 white and 6 from minorities, for a total of 237 individuals) in which MODY was not caused by known MODY genes. Seven chromosomal regions (1q42, 2q24, 2q37, 4p13, 8p23, 11p15, and 19q12) had a parametric heterogeneity logarithm of odds (HLOD) > or =1.00 or a nonparametric logarithm of odds (LOD) > or =0.59 (P < or = 0.05) in the initial screen. After typing additional markers at these loci to reduce the spacing to 2-3 cM, significant linkage was detected on 8p23 (HLOD = 3.37 at D8S1130 and nonparametric LOD = 3.66; P = 2 x 10(-5) at D8S265), where a 4.7-Mb inversion polymorphism is located. Thirty percent of the families (6 of 21) were linked with this region. Another linkage peak on chromosome 2q37 with an HLOD of 1.96 at D2S345/D2S2968 accounted for diabetes in an additional 25% of families (5 of 21). All 6 minority families were among the 11 families linked to these loci. None of the other loci followed up had an HLOD exceeding 1.50. In summary, we have identified a MODY locus on 8p23 that accounts for diabetes in a substantial proportion of MODY cases unlinked to known MODY genes. Another novel MODY locus may be present on 2q37. Cloning these new MODY genes may offer insights to disease pathways that are relevant to the cause of common type 2 diabetes. PMID- 15111510 TI - Complex haplotypes of the PGC-1alpha gene are associated with carbohydrate metabolism and type 2 diabetes. AB - Peroxisome proliferator-activated receptor coactivator-1alpha (PGC-1alpha) is a transcriptional coactivator implicated in transcriptional programs of hepatic gluconeogenesis, oxidative phosphorylation, and insulin release by beta-cells. To study associations of the PGC-1alpha gene locus with carbohydrate metabolism and type 2 diabetes in humans, we identified several polymorphisms in the promoter region that were located in a haplotype block distinct from a second haplotype block containing part of intron 2 and extending beyond exon 13. Each block contained five common haplotypes. Oral glucose tolerance testing revealed associations of promoter haplotype combinations with 30- and 60-min postload plasma glucose levels, whereas haplotypes in both blocks were associated with indexes of beta-cell function. The associations of promoter haplotypes are supported by functional studies showing that some polymorphisms are located in transcription factor binding sites and affect transactivation in an allele specific manner. By comparing patients with type 2 diabetes and control subjects, we observed borderline significant differences of four-loci haplotype distributions in the downstream haplotype block. Moreover, the haplotype that was associated with the strongest insulin response to glucose conferred the lowest risk of type 2 diabetes (P < 0.01). Thus, the PGC-1alpha gene locus influences carbohydrate metabolism and contributes to type 2 diabetes in the population studied. PMID- 15111511 TI - Investigation of the human ANP gene in type 1 diabetic nephropathy: case-control and follow-up studies. AB - The atrial natriuretic peptide gene (PND) is a candidate gene for diabetic nephropathy. We systematically analyzed five nonsynonymous PND polymorphisms and tested the association of genotype and haplotype distributions with diabetic nephropathy in a cross-sectional study and a 6-year follow-up study (489 and 301 type 1 diabetic patients, respectively). For this purpose, a new maximum likelihood method dealing with haplotype-based association analysis for survival data was developed. None of the genotypes or haplotypes were associated with the disease in the case-control study. In the follow-up study, C708T and T2238C showed a weak association with disease progression, but T2238C was strongly associated with progression in poorly controlled subjects (mean HbA(1c) during follow-up was more than the median value [8.5%]; log-rank [TC or CC versus TT], P = 0.007; adjusted hazard ratio, TC or CC versus TT, 2.28, 95% CI 1.10-4.74; P = 0.027). The raw effect of the 2238C allele (hazard risk ratio 1.93, 95% CI 1.15 3.24; P = 0.012) was further confirmed by the haplotype analysis, suggesting that the 2238C allele of PND may affect the course of nephropathy in inadequately controlled type 1 diabetic patients. PMID- 15111512 TI - Genome-wide linkage to chromosome 6 for waist circumference in the Framingham Heart Study. AB - While several loci for BMI have been identified, it is not known whether genes underlie the process of regional fat deposition. We sought to test whether waist circumference, a measure of central adiposity, contains a genetic component. Variance components linkage analysis was performed on 330 families from the Framingham Heart Study original and offspring cohorts, using a 10-cM genome-wide linkage analysis. Overall, 2,086 subjects (51% women), mean age 48 years, were available for analysis. The overall heritability of waist circumference was 0.41. The maximum logarithm of odds (LOD) score in the full dataset was 3.3 on chromosome 6 at marker D6S1009; when subjects were limited to those aged <60 years, the peak LOD score was 3.7 at the same location. Substantial evidence exists for linkage to waist circumference, a measure of central adiposity. Potential candidate genes include ESR1, OPRM1, and NMBR. Further research is necessary to understand the genes involved in central adiposity. PMID- 15111513 TI - Status of research funding by the American Diabetes Association--5th and final year. PMID- 15111514 TI - A direct efficacy and safety comparison of insulin aspart, human soluble insulin, and human premix insulin (70/30) in patients with type 2 diabetes. AB - OBJECTIVE: Because there are limited data on the comparison of insulin aspart and mixed insulin in type 2 diabetes, this trial was performed to compare the efficacy and safety of preprandial insulin aspart with human soluble insulin (HI) and human premix (70% NPH/30% regular) insulin (MIX). RESEARCH DESIGN AND METHODS: A total of 231 type 2 diabetic patients were randomized to insulin aspart (n = 75), HI (n = 80), or MIX (n = 76) for 3 months. Insulin aspart and HI were administered with or without bedtime NPH insulin. A total of 204 patients completed the trial according to protocol. HbA1c, 7-point blood glucose, insulin dosage, and hypoglycemic episodes were recorded. The primary end point was "change of HbA1c" from baseline to last visit. Analysis for equivalence was performed by t tests with three subtests. RESULTS: HbA1c decreased 0.91 +/- 1.00 for insulin aspart, 0.73 +/- 0.87 for HI, and 0.65 +/- 1.10 for MIX with the following confidence intervals: insulin aspart HI (-0.21 to 0.57, P = 0.025), insulin aspart MIX (-0.17 to 0.69, P = 0.092), and HI-MIX (-0.33 to 0.48, P = 0.006). Postprandial blood glucose decreased in the insulin aspart group: 0.44 mmol/l to >1.67 mmol/l compared with HI and 1.1 mmol/l to >1.67 mmol/l compared with MIX. Preprandial insulin doses were similar in the insulin aspart and HI groups (10-14.5 U). Hypoglycemic events per month were 0.56 HI, 0.40 insulin aspart, and 0.19 MIX. CONCLUSIONS: Statistically, insulin aspart was not equivalent to another treatment in terms of HbA1c reduction. Insulin aspart treatment resulted in improved HbA1c and postprandial blood glucose. The application of insulin aspart was safe and well tolerated. PMID- 15111515 TI - Short-term intensive insulin therapy in newly diagnosed type 2 diabetes. AB - OBJECTIVE: Type 2 diabetes is associated with defects in insulin secretion and insulin action. Hyperglycemia may aggravate these defects, a feature known as glucose toxicity. Previous studies have shown that acute correction of hyperglycemia in subjects with long-standing type 2 diabetes gives only short term improvement in glycemic control after discontinuation of insulin. The current study attempts to identify any characteristics of patients with newly diagnosed type 2 diabetes (fasting glucose >11.0 mmol/l) who would have a long term benefit, in terms of glycemic control, from a brief course of insulin therapy. RESEARCH DESIGN AND METHODS: A total of 16 subjects (52 +/- 2 years old [range 36-64], BMI 30.8 +/- 1.9 kg/m2) with newly diagnosed type 2 diabetes had a 2-3 week course of intensive insulin therapy that was then discontinued. RESULTS: Fasting glucose fell from 13.3 +/- 0.7 to 7.0 +/- 0.4 mmol/l, and this improvement was maintained at the 1-year follow-up (6.7 +/- 0.3 mmol/l). The insulin area under the curve for the posttreatment oral glucose tolerance test also improved (8,251 +/- 1,880 before therapy, 18,404 +/- 4,040 directly after insulin therapy, and 42,368 +/- 8,517 pmol.min at the 1-year follow-up). At 1 year, seven of the subjects maintained good glycemic control on diet therapy alone, eight required oral hypoglycemic agent (OHA) therapy, and one required insulin therapy. The distinguishing features of those who did not require OHA or insulin therapy were that they required less insulin during the active insulin therapy phase (0.37 +/- 0.05 vs. 0.73 +/- 0.07 units.kg(-1).day(-1)) and were able to attain a lower fasting serum glucose at the end of the period of insulin therapy (5.9 +/- 0.3 vs. 7.7 +/- 0.4 mmol/l). CONCLUSIONS: These results demonstrate that in newly diagnosed type 2 diabetes with elevated fasting glucose levels, a 2- to 3-week course of intensive insulin therapy can successfully lay a foundation for prolonged good glycemic control. The ease with which normoglycemia is achieved on insulin may predict those patients who can later succeed in controlling glucose levels with attention to diet alone. PMID- 15111516 TI - Gestational diabetes: Is there a relationship between leg length and glucose tolerance? AB - OBJECTIVE: To assess the relationship between leg length and glucose tolerance in pregnancy. RESEARCH DESIGN AND METHODS: The leg length and leg-to-height percentage were prospectively determined on 161 glucose-tolerant women during pregnancy and 61 women with gestational diabetes mellitus (GDM). RESULTS: Women with GDM were a mean of 2.8 cm shorter than women who were glucose tolerant, due entirely to their leg lengths being a mean of 3.2 cm shorter. With respect to the 2-h result on the glucose tolerance test (GTT), there were negative correlations for height (r = -0.161, P = 0.017), leg length (r = -0.266, P < 0.0005), and the leg-to-height percentage (r = -0.294, P < 0.0005). The correlation between the leg-to-height percentage and the 2-h result on the GTT remained significant after adjustment for age (r = -0.252, P < 0.0005) and for age and BMI (r = -0.224, P = 0.001). CONCLUSIONS: Women with GDM are shorter than glucose-tolerant women and have a lower leg-to-height percentage. Consideration of short stature as a risk factor for GDM is not valid without taking into account the leg-to-height percentage. PMID- 15111517 TI - Testosterone and sex hormone-binding globulin predict the metabolic syndrome and diabetes in middle-aged men. AB - OBJECTIVE: In men, hypoandrogenism is associated with features of the metabolic syndrome, but the role of sex hormones in the pathogenesis of the metabolic syndrome and diabetes is not well understood. We assessed the association of low levels of testosterone and sex hormone-binding globulin (SHBG) with the development of the metabolic syndrome and diabetes in men. RESEARCH DESIGN AND METHODS: Concentrations of SHBG and total and calculated free testosterone and factors related to insulin resistance were determined at baseline in 702 middle aged Finnish men participating in a population-based cohort study. These men had neither diabetes nor the metabolic syndrome. RESULTS: After 11 years of follow up, 147 men had developed the metabolic syndrome (National Cholesterol Education Program criteria) and 57 men diabetes. Men with total testosterone, calculated free testosterone, and SHBG levels in the lower fourth had a severalfold increased risk of developing the metabolic syndrome (odds ratio [OR] 2.3, 95% CI 1.5-3.4; 1.7, 1.2-2.5; and 2.8, 1.9-4.1, respectively) and diabetes (2.3, 1.3 4.1; 1.7, 0.9-3.0; and 4.3, 2.4-7.7, respectively) after adjustment for age. Adjustment for potential confounders such as cardiovascular disease, smoking, alcohol intake, and socioeconomic status did not alter the associations. Factors related to insulin resistance attenuated the associations, but they remained significant, except for free testosterone. CONCLUSIONS: Low total testosterone and SHBG levels independently predict development of the metabolic syndrome and diabetes in middle-aged men. Thus, hypoandrogenism is an early marker for disturbances in insulin and glucose metabolism that may progress to the metabolic syndrome or frank diabetes and may contribute to their pathogenesis. PMID- 15111518 TI - Reduction in diabetes-related lower-extremity amputations in The Netherlands: 1991-2000. AB - OBJECTIVE: Lower-extremity amputation is a common complication among patients with diabetes throughout the world. However, few data exist on the actual impact of the recent moves to improve the management of diabetic foot ulcers to reduce the incidence of lower-extremity amputations. The aim was to determine the incidence of lower-extremity amputations among diabetic patients from 1991 to 2000 in The Netherlands. RESEARCH DESIGN AND METHODS: A secondary database containing information regarding all hospital admissions in which a lower extremity amputation occurred for the years 1991-2000 was obtained from the Dutch National Medical Register. Because a patient-unique identifier was included, multiple amputations and hospitalizations for a single individual could be identified. Furthermore, age- and sex-specific diabetes prevalence rates were calculated using a 3-year average for every year, calculating the total diabetic population in the Netherlands at risk for every year. RESULTS: In 1991, a total of 1,687 patients with diabetes had been admitted 1,865 times for 2,409 amputations. In 2000, a total of 1,673 patients with diabetes were admitted 1,932 times for 2,448 amputations. The overall incidence rates of the number of patients who underwent lower-extremity amputation decreased over the years from 55.0 to 36.3 per 10,000 patients with diabetes (P < 0.05). Both in men (71.8 vs. 46.1, P < 0.05) and women (45.0 vs. 28.0, P < 0.05) with diabetes, a significant decrease could be observed. Mean duration of hospitalization decreased from 45.0 days (SD 44.4) in 1991 to 36.2 days (SD 38.4) in 2000; decreases were observed for both men and women. CONCLUSIONS: Over the years observed in this study, the incidence rates of diabetes-related lower-extremity amputation in The Netherlands was found to decrease in both men (36%) and women (38%) with diabetes. Furthermore, the duration of hospitalization decreased over time. PMID- 15111519 TI - Global prevalence of diabetes: estimates for the year 2000 and projections for 2030. AB - OBJECTIVE: The goal of this study was to estimate the prevalence of diabetes and the number of people of all ages with diabetes for years 2000 and 2030. RESEARCH DESIGN AND METHODS: Data on diabetes prevalence by age and sex from a limited number of countries were extrapolated to all 191 World Health Organization member states and applied to United Nations' population estimates for 2000 and 2030. Urban and rural populations were considered separately for developing countries. RESULTS: The prevalence of diabetes for all age-groups worldwide was estimated to be 2.8% in 2000 and 4.4% in 2030. The total number of people with diabetes is projected to rise from 171 million in 2000 to 366 million in 2030. The prevalence of diabetes is higher in men than women, but there are more women with diabetes than men. The urban population in developing countries is projected to double between 2000 and 2030. The most important demographic change to diabetes prevalence across the world appears to be the increase in the proportion of people >65 years of age. CONCLUSIONS: These findings indicate that the "diabetes epidemic" will continue even if levels of obesity remain constant. Given the increasing prevalence of obesity, it is likely that these figures provide an underestimate of future diabetes prevalence. PMID- 15111520 TI - Diabetes and impaired fasting glycemia in the tribes of Khagrachari hill tracts of Bangladesh. AB - OBJECTIVE: To determine the prevalence of type 2 diabetes and impaired fasting glycemia (IFG) in a tribal population of Bangladesh. RESEARCH DESIGN AND METHODS: A cluster sampling of 1,287 tribal subjects of age > or =20 years was investigated. They live in a hilly area of Khagrachari in the far northeast of Bangladesh. Fasting plasma glucose, blood pressure, height, weight, waist girth, and hip girth were measured. Lipid fractions were also estimated. We used the 1997 American Diabetes Association diagnostic criteria. RESULTS: The crude prevalence of type 2 diabetes was 6.6% and IFG was 8.5%. The age-standardized (20 70 years) prevalence of type 2 diabetes (95% CI) was 6.4% (4.96-7.87) and of IFG was 8.4% (6.48-10.37). Both tribesmen and women had equal risk for diabetes and IFG. Compared with the lower-income group, the participants with higher income had a significantly higher prevalence of type 2 diabetes (18.8 vs. 3.1%, P < 0.001) and IFG (17.2 vs. 4.3%, P < 0.001). Using logistic regression, we found that increased age, high-income group, and increased central obesity were the important risk factors of diabetes. CONCLUSIONS: The prevalence of diabetes in the tribal population was higher than that of the nontribal population of Bangladesh. Older age, higher central obesity, and higher income were proven significant risk factors of diabetes. High prevalence of diabetes among these tribes indicates that the prevalence of diabetes and its complications will continue to increase. Evidently, health professionals and planners should initiate diabetes care in these tribal communities. PMID- 15111521 TI - Are low-income elderly patients at risk for poor diabetes care? AB - OBJECTIVE: Diabetes is common among low-income elderly, dual-eligible (DE) Medicare/Medicaid patients resulting in significant morbidity, mortality, and cost. However, the quality of diabetes care delivered to these patients has not been evaluated. The aims of this study were to describe the quality of diabetes care provided to DE patients and compare it with non-DE patients. RESEARCH DESIGN AND METHODS: This was a cross-sectional analysis of administrative claims from 1 January 1997 through 31 December 1998. A total of 9,453 patients aged 65-75 years with diabetes participated in the study. These were Colorado Medicare fee-for service (FFS) outpatients. The main outcome measures consisted of a proportion of patients receiving an annual hemoglobin A1c test, biennial eye examination, biennial lipid test, and all three of these care processes. RESULTS: The mean patient age was 71 +/- 2.8 years. Over 22% of patients were identified as dual eligible, and they were significantly more likely to be younger, female, and of minority race/ethnicity; reside in a rural location; and have comorbid conditions compared with the non-DE population. DE patients had more visits to primary care physicians, emergency departments, and hospitalizations but were less likely to visit endocrinologists. DE patients were significantly less likely to receive an annual A1c test (73 vs. 81%; P < 0.0001), biennial ophthalmologic examination (63 vs. 75%; P < 0.0001), and biennial lipid testing (43 vs. 57%; P < 0.0001). The adjusted odds ratio of urban DE patients receiving all three care measures was 0.60 (95% CI 0.52-0.69) compared with urban non-DE patients. Minority race/ethnicity and emergency department use were significantly associated with not receiving diabetes care, whereas endocrinology visits were associated with an increased odds of receiving diabetes care. CONCLUSIONS: DE Medicare/Medicaid status was independently associated with not receiving diabetes care, especially among those in urban areas. PMID- 15111522 TI - Diabetes, depression, and quality of life: a population study. AB - OBJECTIVE: The aim of the study was to assess the prevalence of diabetes and depression and their associations with quality of life using a representative population sample. RESEARCH DESIGN AND METHODS: The study consisted of a representative population sample of individuals aged > or = 15 years living in South Australia comprising 3,010 personal interviews conducted by trained health interviewers. The prevalence of depression in those suffering doctor-diagnosed diabetes and comparative effects of diabetic status and depression on quality-of life dimensions were measured. RESULTS: The prevalence of depression in the diabetic population was 24% compared with 17% in the nondiabetic population. Those with diabetes and depression experienced an impact with a large effect size on every dimension of the Short Form Health-Related Quality-of-Life Questionnaire (SF-36) as compared with those who suffered diabetes and who were not depressed. A supplementary analysis comparing both depressed diabetic and depressed nondiabetic groups showed there were statistically significant differences in the quality-of-life effects between the two depressed populations in the physical and mental component summaries of the SF-36. CONCLUSIONS: Depression for those with diabetes is an important comorbidity that requires careful management because of its severe impact on quality of life. PMID- 15111523 TI - Treatment of diabetic gastroparesis by high-frequency gastric electrical stimulation. AB - OBJECTIVE: To investigate the long-term efficacy of high-frequency gastric electrical stimulation (GES) for treating diabetic gastroparesis. RESEARCH DESIGN AND METHODS: This is a retrospective review of 48 adult diabetic patients with refractory gastroparesis who had a GES system implanted surgically and had follow up evaluations at 6 and 12 months. The outcome measures were total symptom score (TSS), derived from six upper gastrointestinal (GI) symptom subscores; health related quality of life (HQOL), including physical composite score (PCS) and mental composite score (MCS) assessed by SF-36 questionnaire, radionuclide gastric emptying test, nutritional status, HbA1c, and adverse events. RESULTS: In comparison with baseline, TSS, all six upper GI symptom subscores, PCS, and MCS were significantly improved at 6 months, with the improvement sustained at 12 months. Of 13 patients receiving nutritional support at baseline by tube feeding, only 5 required supplemental enteral feeding at 12 months, and none of the 9 on total parenteral nutrition continued this support. The mean number of hospitalization days during the year after GES was significantly reduced by 52 days compared with the prior year. HbA1c levels were significantly reduced at 12 months. Gastric emptying was only minimally and not significantly faster. Because of infections at the pulse generator pocket site, four patients had their GES systems removed 3-17 months postsurgery. CONCLUSIONS: In diabetic patients with refractory gastroparesis, high-frequency GES by a permanently implanted system significantly improved upper GI symptoms, HQOL, nutritional status, glucose control, and hospitalizations with an acceptably low complication rate. PMID- 15111524 TI - Orlistat augments postprandial increases in glucagon-like peptide 1 in obese type 2 diabetic patients. AB - OBJECTIVE: Orlistat leads to improved glycemic control in obese type 2 diabetic patients, which is attributed to decreased insulin resistance associated with weight loss. Glucagon-like peptide 1 (GLP-1) and glucose-dependent insulinotropic peptide (GIP) are gut hormones that are secreted in response to food intake, and they both stimulate insulin secretion. Orlistat decreases fat absorption and increases intestinal fat content, which may lead to increased secretion of these peptides. In this pilot study, we tested the hypothesis that increased levels of these intestinal hormones may be involved in the improvement of postprandial hyperglycemia observed previously with orlistat in type 2 diabetic patients. RESEARCH DESIGN AND METHODS: A total of 29 type 2 diabetic patients, who were not taking insulin or alpha-glucosidase inhibitors, were enrolled in the study. On a crossover and single-blind design, after an overnight fasting, the patients received 120-mg orlistat or placebo capsules, followed by a standard 600-kcal mixed meal that contained 38% fat. At baseline and 60 min after the meal, blood samples were obtained for the measurement of GLP-1, GIP, insulin, C-peptide, triglycerides, free fatty acids, and glucose. RESULTS: All measured parameters increased significantly in response to the mixed meal compared with baseline, both with orlistat or placebo. When compared with the placebo, the orlistat administration resulted in a significantly enhanced postprandial increase in GLP 1 and C-peptide levels and attenuated the postprandial rise in glucose and triglycerides. CONCLUSIONS: The results of this study suggest that apart from decreasing insulin resistance as a result of weight loss, orlistat may increase postprandial GLP-1 levels, thereby enhancing the insulin secretory response to the meal and blunting the postprandial rise in glucose in type 2 diabetic patients. Increased GLP-1 levels, which lead to decreased food intake, may also contribute to the weight loss that is associated with the use of this drug. PMID- 15111525 TI - Insulin detemir offers improved glycemic control compared with NPH insulin in people with type 1 diabetes: a randomized clinical trial. AB - OBJECTIVE: Insulin detemir is a soluble long-acting basal insulin analog designed to overcome the limitations of conventional basal insulin formulations. Accordingly, insulin detemir has been compared with NPH insulin with respect to glycemic control (HbA1c, prebreakfast glucose levels and variability, and hypoglycemia) and timing of administration. RESEARCH DESIGN AND METHODS: People with type 1 diabetes (n = 408) were randomized in an open-label, parallel-group trial of 16-week treatment duration using either insulin detemir or NPH insulin. Insulin detemir was administered twice daily using two different regimens, either before breakfast and at bedtime (IDet(morn+bed)) or at a 12-h interval (IDet(12h)). NPH insulin was administered before breakfast and at bedtime. Mealtime insulin was given as the rapid-acting insulin analog insulin aspart. RESULTS: With both insulin detemir groups, clinic fasting plasma glucose was lower than with NPH insulin (IDet(12h) vs. NPH, -1.5 mmol/l [95% CI -2.51 to 0.48], P = 0.004; IDet(morn+bed) vs. NPH, -2.3 mmol/l (-3.32 to -1.29), P < 0.001), as was self-measured prebreakfast plasma glucose (P = 0.006 and P = 0.004, respectively). The risk of minor hypoglycemia was lower in both insulin detemir groups (25%, P = 0.046; 32%, P = 0.002; respectively) compared with NPH insulin in the last 12 weeks of treatment, this being mainly attributable to a 53% reduction in nocturnal hypoglycemia in the IDet(morn+bed) group (P < 0.001). Although HbA1c for each insulin detemir group was not different from the NPH group, HbA1c for the pooled insulin detemir groups was significantly lower than for the NPH group (mean difference -0.18% [-0.34 to -0.02], P = 0.027). Within person between-day variation in self-measured prebreakfast plasma glucose was lower for both detemir groups (both P < 0.001). The NPH group gained weight during the study, but there was no change in weight in either of the insulin detemir groups (IDet(12h) vs. NPH, -0.8 kg [-1.44 to -0.24], P = 0.006; IDet(morn+bed) vs. NPH, -0.6 kg [-1.23 to -0.03], P = 0.040). CONCLUSIONS: Overall glycemic control with insulin detemir was improved compared with NPH insulin. The data provide a basis for tailoring the timing of administration of insulin detemir to the individual person's needs. PMID- 15111526 TI - Telecare for patients with type 1 diabetes and inadequate glycemic control: a randomized controlled trial and meta-analysis. AB - OBJECTIVE: To determine the efficacy of telecare (modem transmission of glucometer data and clinician feedback) to support intensive insulin therapy in patients with type 1 diabetes and inadequate glycemic control. RESEARCH DESIGN AND METHODS: Thirty-one patients with type 1 diabetes on intensive insulin therapy and with HbA1c >7.8% were randomized to telecare (glucometer transmission with feedback) or control (glucometer transmission without feedback) for 6 months. The primary end point was 6-month HbA1c. To place our findings in context, we pooled HbA1c change from baseline reported in randomized trials of telecare identified in a systematic review of the literature. RESULTS: Compared with the control group, telecare patients had a significantly lower 6-month HbA1c (8.2 vs. 7.8%, P = 0.03, after accounting for HbA1c at baseline) and a nonsignificant fourfold greater chance of achieving 6-month HbA1c < or =7% (29 vs. 7%; risk difference 21.9%, 95% CI -4.7 to 50.5). Nurses spent 50 more min/patient giving feedback on the phone with telecare patients than with control patients. Meta-analysis of seven randomized trials of adult patients with type 1 diabetes found a 0.4% difference (95% CI 0-0.8) in HbA1c mean change from baseline between the telecare and control groups. CONCLUSIONS: Telecare is associated with small effects on glycemic control in patients with type 1 diabetes on intensive insulin therapy but with inadequate glycemic control. PMID- 15111527 TI - Cost-effectiveness analysis of telemedicine to evaluate diabetic retinopathy in a prison population. AB - OBJECTIVE: A cost-effectiveness analysis was conducted to investigate the clinical and economic impact of teleophthalmology in evaluating diabetic retinopathy in prison inmates with type 2 diabetes. RESEARCH DESIGN AND METHODS: Based on a hypothetical teleophthalmology system to evaluate diabetic retinopathy patients with type 2 diabetes in a prison care setting, a Markov decision model was developed with probability and cost data derived primarily from published epidemiological and outcome studies. A 40-year-old African-American man with type 2 diabetes was used as a reference case subject. The number of quality-adjusted life-years (QALYs) gained was used as the clinical outcome, and the cost in U.S. dollars from the year 2003 was used as the economic outcome. Teleophthalmology and nonteleophthalmology strategies were compared using an expected QALYs calculation and two types of sensitivity analyses: probabilistic and traditional n-way sensitivity analyses. RESULTS: The teleophthalmology strategy dominates in the cost-effectiveness analysis for the reference case subject: 16,514/18.73 dollars QALYs for teleophthalmology and 17,590/18.58 dollars QALYs for nonteleophthalmology. Ninety percent of the Monte Carlo simulations showed cost effectiveness (annual cost/QALYs < or = 50,000 dollars) in the teleophthalmology strategy based on an assumed inmate population. Teleophthalmology is the better strategy if the number of diabetic inmates in the prison community is >500. CONCLUSIONS: Our cost-effectiveness analysis demonstrates that teleophthalmology holds great promise to reduce the cost of inmate care and reduce blindness caused by diabetic retinopathy in type 2 diabetic patients. PMID- 15111528 TI - Contrasting diabetes phenotypes associated with hepatocyte nuclear factor-1alpha and -1beta mutations. AB - OBJECTIVE: Mutations in the highly homologous transcription factors hepatocyte nuclear factor (HNF)-1alpha and -1beta cause maturity-onset diabetes of the young types 3 and 5, respectively. Diabetes due to HNF-1alpha mutations is well characterized. However, physiological assessment of the HNF-1beta phenotype is limited. We aimed to test the hypothesis that the diabetes phenotype due to HNF 1beta mutations is similar to that in HNF-1alpha. RESEARCH DESIGN AND METHODS: Fasting biochemistry and a tolbutamide-modified intravenous glucose tolerance test (IVGTT) were compared in matched HNF-1beta, HNF-1alpha, type 2 diabetic, and control subjects. Homeostasis model assessment indexes were determined from fasting insulin and glucose. The peak measures for the insulin increment after tolbutamide and for the insulin increment after glucose were determined from the IVGTT. RESULTS: The HNF-1beta patients showed a 2.4-fold reduction in insulin sensitivity compared with the HNF-1alpha patients (P = 0.001) with fasting insulin concentrations 2.7-fold higher (P = 0.004). HNF-1beta patients had lower HDL cholesterol (1.17 vs. 1.46 mmol/l; P = 0.009) and higher triglyceride (2.2 vs. 1.35 mmol/l; P = 0.015) levels than HNF-1alpha patients. The HNF-1beta patients had similar beta-cell responses to tolbutamide and glucose as the type 2 diabetic patients, but in the HNF-1alpha patients, the tolbutamide response was considerably increased relative to the response to glucose (P = 0.002). CONCLUSIONS: HNF-1beta patients have a different diabetes phenotype than HNF 1alpha patients. Those with HNF-1beta mutations have hyperinsulinemia and associated dyslipidemia consistent with insulin resistance and may have a different beta-cell defect. This suggests that despite considerable homology and a shared binding site, HNF-1alpha and HNF-1beta have a different role in maintaining normal glucose homeostasis. This result suggests a new etiological pathway for insulin resistance involving HNF-1beta. PMID- 15111529 TI - Adult-onset atypical (type 1) diabetes: additional insights and differences with type 1A diabetes in a European Mediterranean population. AB - OBJECTIVE: In 1997, the American Diabetes Association proposed two subcategories for type 1 diabetes: type 1A or immunomediated diabetes and type 1B or idiopathic diabetes characterized by negative beta-cell autoimmunity markers, lack of association with HLA, and fluctuating insulinopenia. The aim of this study was to examine clinical characteristics, beta-cell function, HLA typing, and mutations in maturity-onset diabetes of the young (MODY) genes in patients with atypical type 1 diabetes (type 1 diabetes diagnosed at onset, without pancreatic autoantibodies and fluctuating insulinopenia). RESEARCH DESIGN AND METHODS: Eight patients with atypical type 1 diabetes (all men, 30.7 +/- 7.6 years) and 16 newly diagnosed age- and sex-matched patients with type 1A diabetes were studied retrospectively. Islet cell, GAD, tyrosine phosphatase and insulin antibodies, and basal and stimulated plasma C-peptide were measured at onset and after 1 year. HLA-DRB1-DQA1-DQB1 typing and screening for mutations in the HNF-1alpha and HNF-4alpha genes were performed from genomic DNA. RESULTS: Atypical patients displayed significantly higher BMI and better beta-cell function at onset and after 12 months. Three patients carried protective or neutral type 1 diabetes haplotypes, five patients displayed heterozygosity for susceptible and protective haplotypes, and seven patients showed Asp(beta57). We found a nondescribed variant Pro436Ser in exon 10 of the HNF-4alpha gene in one atypical patient without susceptible haplotypes. CONCLUSIONS: In our population, there are atypical forms of young adult-onset ketosis-prone diabetes initially diagnosed as type 1 diabetes, differing from type 1 diabetes in the absence of beta-cell autoimmunity, persistent beta-cell function capacity, fluctuating insulin requirements and ketosis-prone episodes, as well as clinical features of type 2 diabetes. Only one subgroup could be strictly classified as having type 1B diabetes. Additional information is still needed to improve our understanding of the mechanisms that finally lead to the disease. PMID- 15111530 TI - Reduced coronary artery and abdominal aortic calcification in Hispanics with type 2 diabetes. AB - OBJECTIVE: To compare lifestyle factors, cardiovascular risk factors, and coronary artery calcium (CAC) and abdominal aortic calcium (AAC) levels in Hispanic and non-Hispanic white (NHW) individuals with type 2 diabetes. RESEARCH DESIGN AND METHODS: We recently demonstrated in a small group of nonreferred, healthy, nondiabetic subjects that CAC was reduced in Hispanics compared with NHWs, despite a worse cardiovascular risk factor profile. In this study, we evaluated whether this ethnic disparity in vascular calcification was present in individuals with type 2 diabetes and in several different arterial beds. Hispanic and NHW subjects (n = 245) with type 2 diabetes were evaluated for cardiovascular risk factors using questionnaires and assays of plasma biomarkers. CAC and AAC were measured by electron-beam computer-assisted tomography. RESULTS: Although Hispanics were slightly younger than NHWs, other standard risk factors and novel cardiovascular risk factors, including plasminogen activator-1 and fibrinogen levels, were similar between the groups. Despite the similar risk factor profile, the prevalence of cardiovascular disease (CVD) and mean and median levels of CAC and AAC were lower in Hispanics. Furthermore, the distribution of these calcium scores differed from that of NHWs (P < 0.05), with significantly fewer Hispanic subjects having high CAC or AAC scores. These differences were not explained by differences in CVD prevalence or any measured lifestyle or risk factor. CONCLUSIONS: Hispanics with type 2 diabetes have reduced CAC and AAC levels compared with NHW subjects, suggesting a reduction in the overall burden of vascular calcification and atherosclerosis. These data are consistent with the notion that Hispanics are protected against the development of CVD. PMID- 15111531 TI - Increasing severity of cardiovascular risk factors with increasing middle cerebral artery stenotic involvement in type 2 diabetic Chinese patients with asymptomatic cerebrovascular disease. AB - OBJECTIVE: To identify determinants associated with increasing severity of middle cerebral artery (MCA) stenosis in asymptomatic Chinese type 2 diabetic patients with and without MCA stenosis determined using transcranial Doppler. Conventional risk factors contribute to the pathogenesis of ischemic stroke, and differences in the pattern of these may explain the heterogeneity of disease presentation in different populations. In Chinese patients, MCA stenosis is the most commonly identified intracranial vascular lesion. RESEARCH DESIGN AND METHODS: Anthropometric and fasting biochemical parameters were compared between type 2 diabetic patients with MCA stenosis in one (n = 185) or both (n = 200) vessels and 1,492 type 2 diabetic patients without evidence of stenosis. RESULTS: Increasing MCA stenotic vascular involvement was associated with significantly increasing age, duration of diabetes, systolic blood pressure, and LDL cholesterol, but with lower glucose levels. There was also an increased prevalence of hypertension, dyslipidemia, and use of blood pressure-and glucose lowering agents in the patients with MCA stenosis. Concomitant significant increases in the prevalence of peripheral vascular disease and retinopathy were also observed in the patients with MCA stenosis. CONCLUSIONS: Transcranial Doppler examination identified stenosis in one or both MCAs in over one-fifth of the Chinese type 2 diabetic subjects without symptoms of cerebrovascular disease. A number of conventional cardiovascular risk factors were closely associated with MCA stenosis. This technique may allow the identification of a particularly high risk group, and further studies are required to determine whether asymptomatic MCA stenosis is predictive of primary cerebrovascular events and whether intensive treatment of risk factors would reduce the risk. PMID- 15111532 TI - Influence of caffeine on heart rate variability in patients with long-standing type 1 diabetes. AB - OBJECTIVE: The effect of caffeine on cardiovascular health remains controversial. Patients with long-standing type 1 diabetes are at risk of autonomic failure and sudden cardiac death. We investigated the effects of caffeine on autonomic dysfunction (as assessed by heart rate variability [HRV]) in this high-risk group and in a control population. RESEARCH DESIGN AND METHODS: Using a randomized blinded, placebo-controlled, crossover design trial, we examined 2 weeks of caffeine consumption (250 mg twice daily) on HRV in 20 type 1 diabetic patients and 10 matched healthy volunteers. RESULTS: Baseline HRV was blunted in the diabetic patients (P < 0.0005 vs. control subjects) and markedly increased by caffeine in both groups (+103% in the group with diabetes [P = 0.009] and +38% in control subjects [P = 0.002]). The caffeine-associated increase in HRV was not statistically different between the control and the type 1 diabetes groups (P = 0.16). CONCLUSIONS: Modest amounts of caffeine improved autonomic function in diabetic patients and healthy volunteers. For individuals with abnormal HRV, regular caffeine use may have the potential to reduce the risk of cardiovascular events. PMID- 15111533 TI - An explanation for the increase in heart disease mortality rates in diabetic Pima Indians: effect of renal replacement therapy. AB - OBJECTIVE: Diabetic nephropathy (DN) became the leading cause of death in diabetic Pima Indians in the 1970s, but was superseded by ischemic heart disease (IHD) in the 1980s. This study tests the hypothesis that the rise in the IHD death rate between 1965 and 1998 is attributable to access to renal replacement therapy (RRT). RESEARCH DESIGN AND METHODS: Underlying causes of death were determined among 2,095 diabetic Pima Indians > or = 35 years old during four 8.5 year time intervals. To assess the effect of access to RRT on IHD death rates, trends were reexamined after subjects receiving RRT were classified as if they had died of DN. RESULTS: During a median follow-up of 11.1 years (range 0.01-34), 818 subjects died. The age- and sex-adjusted DN death rate decreased over the 34 year study (P = 0.05), whereas the IHD death rate increased from 3.3 deaths/1,000 person-years (95% CI 1.4-5.2) to 6.3 deaths/1,000 person-years (95% CI 4.5-8.0; P = 0.03). After 151 subjects on RRT were reclassified as if they had died of DN, the death rate for DN increased from 4.8 deaths/1,000 person-years (95% CI 2.6-7) to 11.3 deaths/1,000 person-years (95% CI 9-13.6; P = 0.0007), whereas the increase in the IHD death rate disappeared (P = 0.57). CONCLUSIONS: The incidence rate of renal failure attributable to diabetes has increased rapidly over the past 34 years in Pima Indians. IHD has emerged as the leading cause of death due largely to the availability of RRT and to changes in the pattern of death among those with DN. PMID- 15111534 TI - Effect of diabetes on long-term mortality following contemporary percutaneous coronary intervention: analysis of 4,284 cases. AB - OBJECTIVE: Diabetic patients are known to have reduced long-term survival following percutaneous transluminal coronary angioplasty compared with nondiabetic patients. However, it is unknown whether this survival disadvantage has persisted in the era of contemporary percutaneous coronary intervention (PCI) techniques, which include the widespread use of stents and the availability of platelet glycoprotein (GP) IIb/IIIa inhibitors. RESEARCH DESIGN AND METHODS: Three hospitals in New York City contributed prospectively defined data on 4,284 patients undergoing PCI. The primary end point was all-cause mortality following hospital discharge for PCI. RESULTS: Hypertension, renal insufficiency, and renal failure requiring dialysis were all more common in diabetic patients, whereas active smoking was less frequent. Congestive heart failure on admission was more common in diabetic than nondiabetic patients (7.7 vs. 4.0%, P < 0.001). Stents were placed in 78% of nondiabetic patients and 75% of diabetic patients (P = 0.045). Platelet GP IIb/IIIa antagonists were administered to 23% of nondiabetic and 24% of diabetic patients (P = NS). At a mean follow-up of 3 years, mortality was 8% among nondiabetic patients and 13% for diabetic patients (P < 0.001). After adjustment for differences in baseline characteristics between nondiabetic and diabetic patients, diabetes remained a significant independent hazard for late mortality (hazard ratio 1.462, 95% CI 1.169-1.828; P = 0.001). CONCLUSIONS: Following contemporary PCI, diabetic patients continue to have worse survival than nondiabetic patients. PMID- 15111535 TI - Risk of community-acquired pneumococcal bacteremia in patients with diabetes: a population-based case-control study. AB - OBJECTIVE: We conducted this population-based case-control study to examine whether diabetes is associated with an increased risk of community-acquired pneumococcal bacteremia. RESEARCH DESIGN AND METHODS: We included 598 cases in the North Jutland County Bacteremia Registry, Denmark, with residence in the county and a first hospitalization for community-acquired pneumococcal bacteremia from 1992 through 2001. Ten sex- and age-matched population control subjects per case were selected, using a unique personal identifier. Diabetes was determined by record linkage with the County Prescription Database (for prescriptions for antidiabetic drugs) and the Hospital Discharge Registry (for previous hospitalizations with diabetes or diabetic complications). We performed conditional logistic regression to estimate odds ratios (ORs) for pneumococcal bacteremia among diabetic and nondiabetic persons, with adjustment for a range of comorbid diseases considered to be risk factors for pneumococcal infection. RESULTS: The crude OR for pneumococcal bacteremia in persons with diabetes was 1.9 (95% CI 1.4-2.6). After adjustment for comorbidity, the OR decreased to 1.5 (95% CI 1.1-2.0). The impact of diabetes on the risk for pneumococcal bacteremia was most pronounced in adults aged 40 years and younger (adjusted OR 4.2, 95% CI 1.1-16.7) and in persons without any other coexisting morbidity (adjusted OR 2.3, 95% CI 1.3-3.9). Under the assumptions that the association was causal and that there is a 5% overall prevalence of diabetes in our study population, 24 of 1,000 admissions with incident pneumococcal bacteremia may be attributed to diabetes. CONCLUSIONS: Diabetes seems to be a risk factor for community-acquired pneumococcal bacteremia. PMID- 15111536 TI - Insulin sensitivity in the offspring of women with type 1 and type 2 diabetes. AB - OBJECTIVE: To determine if insulin sensitivity is altered in prepubertal offspring exposed to a diabetic intrauterine environment. RESEARCH DESIGN AND METHODS: Fifteen control children, 17 offspring of type 1 diabetic women, and 10 offspring of type 2 diabetic women, aged between 5 and 10 years, underwent a frequently sampled intravenous glucose tolerance test (FSIGTT). Weight and height were measured, and body composition was calculated using bioelectrical impedance. Bergman's minimal model was applied to the glucose and insulin measurements to obtain values for insulin sensitivity (Si), acute insulin response (AIR), and glucose effectiveness (Sg). RESULTS: Si was lowest in the offspring of type 2 diabetic mothers, and AIR was highest in this group, although neither of these changes reached significance (Si, P = 0.2, and AIR, P = 0.3). Offspring of type 2 diabetic mothers had higher BMI SD scores (P = 0.004) and percentage fat mass (P = 0.002) than the children in the other two groups. The BMI SD score and percentage fat mass in the subjects, as well as maternal insulin dose, were negatively correlated with offspring insulin sensitivity. CONCLUSIONS: Intrauterine exposure to hyperglycemia by itself was not associated with alterations in glucose regulation in prepubertal offspring. Children of mothers with type 2 diabetes, however, were overweight, and they had a tendency for a reduced Si. The combined effect of genetic and postnatal environmental factors, rather than prenatal exposure to hyperglycemia, may place this group at risk for developing impaired glucose tolerance in later life. PMID- 15111537 TI - Natural progression of diabetic peripheral neuropathy in the Zenarestat study population. AB - OBJECTIVE: The aim of this study was to report the baseline and natural progression of diabetic peripheral neuropathy over 12 months in a large mild-to moderate neuropathy population. RESEARCH DESIGN AND METHODS: Patients from a multicentered trial of zenarestat, an aldose reductase inhibitor, had serial measures of neurologic function, including nerve conduction studies (NCSs), quantitative sensory testing (QST), and clinical neuropathy rating scores at baseline and at 12 months. Baseline population descriptors and changes in neurologic function in placebo-treated patients were analyzed. RESULTS: Sural sensory velocity (P = 0.0008 [95% CI -1.04 to -0.27]), median sensory amplitude (P = 0.0021 [-1.3 to -0.29]), median distal motor latency (P = 0.002 [0.09 0.28]), cool thermal QST (P = 0.0005 [0.27-0.94]), and Michigan Neuropathy Screening Instrument results (P = 0.0087 [0.04-0.30]) declined significantly from baseline in the placebo population. NCS changes from baseline were independent of baseline HbA1c stratification. CONCLUSIONS: The neurologic decline over 12 months is evident when measured by NCS and cool thermal QST. Other measures (vibration QST, neuropathy rating scores, monofilament examination) are insensitive to changes over 12 months in a mild-to-moderate affected population of this size. PMID- 15111538 TI - Sural nerve sorbitol in patients with diabetic sensorimotor polyneuropathy. AB - OBJECTIVE: Nerve sorbitol levels have been measured in sural nerve biopsy samples from patients with diabetic sensorimotor polyneuropathy in several studies using different methods and measurement units. In this study, we compared the results of sorbitol assays to determine the required sensitivity of analytical methods for nerve sorbitol measurements. RESEARCH DESIGN AND METHODS: We performed a literature search using PaperChase for reports of nerve sorbitol in diabetic patients and selected those with nerve conduction studies to delineate the severity of nerve damage. RESULTS: In patients who had undergone a nerve conduction study, the standardized nerve sorbitol levels were 0.034-0.300 nmol/mg wet nerve. CONCLUSIONS: Our results showed the level of sensitivity required in laboratory methodology to perform this assay in the target population and aid in the planning of clinical research trials of aldose reductase inhibitor agents. PMID- 15111539 TI - Connective tissue growth factor is increased in plasma of type 1 diabetic patients with nephropathy. AB - OBJECTIVE: Connective tissue growth factor (CTGF) is strongly upregulated in fibrotic disorders and has been hypothesized to play a role in the development and progression of diabetes complications. The aim of the present study was to investigate the possible association of plasma CTGF levels in type 1 diabetic patients with markers relevant to development of diabetes complications. RESEARCH DESIGN AND METHODS: Plasma CTGF levels (full-length and NH2-terminal fragments) were determined in 62 well-characterized patients with type 1 diabetes and in 21 healthy control subjects. Correlations of these plasma CTGF levels with markers of glycemic control, platelet activation, endothelial activation, nephropathy, and retinopathy were investigated. RESULTS: -Elevated plasma NH2-terminal fragment of CTGF (CTGF-N) levels were detected in a subpopulation of type 1 diabetic patients and were associated with diabetic nephropathy. Stepwise regression analysis revealed contribution of albuminuria, creatinine clearance, and duration of diabetes as predictors of plasma CTGF-N level. Elevation of plasma CTGF-N levels in patients with retinopathy was probably due to renal comorbidity. CONCLUSIONS: Plasma CTGF-N levels are elevated in type 1 diabetic patients with nephropathy and appear to be correlated with proteinuria and creatinine clearance. Further studies will be needed to determine the relevance of plasma CTGF as a clinical marker and/or pathogenic factor in diabetic nephropathy. PMID- 15111540 TI - High prevalence of glucose abnormalities in patients with hepatitis C virus infection: a multivariate analysis considering the liver injury. AB - OBJECTIVE: The aim of this study was to compare the prevalence of both impaired fasting glucose (IFG) and diabetes between hepatitis C virus (HCV)-infected patients and patients with other liver diseases but anti-HCV-, taking into account the degree of liver damage. RESEARCH DESIGN AND METHODS: A total of 642 consecutive patients attending the outpatient liver unit of a university hospital (498 anti-HCV+ and 144 anti-HCV-) were prospectively recruited. Patients were classified as having chronic hepatitis (n = 472) or cirrhosis (n = 170) by means of the result of either a liver biopsy or by typical clinical features. A logistic regression model was used to determine independent associations of covariates (age, sex, BMI, HCV antibody status, and triglycerides) with the presence of glucose abnormalities. RESULTS: A threefold increase in the prevalence of glucose abnormalities was observed in HCV+ patients with chronic hepatitis in comparison with HCV- subjects (32 vs. 12%; P = 0.0003). In contrast, among patients with cirrhosis, although both diabetes and IFG were more prevalent in anti-HCV+ patients (40%) than in anti-HCV- patients (36%), the differences were not statistically significant. Finally, the logistic regression analysis showed that HCV infection was independently related to glucose abnormalities in those patients with chronic hepatitis (odds ratio 4.26 [95% CI 2.03-8.93]). In contrast, HCV was not an independent predictor of glucose abnormalities in cirrhotic patients. CONCLUSIONS: The high prevalence of IFG and diabetes found in HCV-infected patients observed in our study suggests that screening for glucose abnormalities should be indicated in these patients. In addition, we provide evidence that the genuine connection between HCV infection and diabetes is initiated at early stages of hepatic disease. PMID- 15111541 TI - Parallel increase in urinary excretion rates of immunoglobulin G, ceruloplasmin, transferrin, and orosomucoid in normoalbuminuric type 2 diabetic patients. AB - OBJECTIVE: Increased urinary excretions of several plasma proteins with different molecular radii <55 A and different isoelectric points (pI), such as IgG, ceruloplasmin, transferrin, and orosomucoid, have been independently reported to precede the development of microalbuminuria in diabetic patients. We examined whether increases in urinary excretions of these proteins would be in parallel in the same patient. RESEARCH DESIGN AND METHODS: Urinary excretion rates of proteins mentioned above in timed overnight urine samples were evaluated in 61 normoalbuminuric type 2 diabetic patients (group D) aged 40-60 years and in 17 age-matched control subjects (group C). RESULTS: The excretion rates of these proteins were significantly higher in group D than in group C. These exhibited a strong linear correlation with each other and had a weak correlation with the excretion rate of N-acethylglucosaminidase. The excretion rate of alpha2 macroglobulin with large molecular radii of 88 A was not different between groups C and D, nor did they have any correlations with the excretion rates of the other proteins. Creatinine clearance and blood pressure levels in group D were significantly higher than those in group C. CONCLUSIONS: In normoalbuminuric diabetic patients, excretion rates of plasma proteins with molecular radii <55 A increased in parallel with each other. In view of our previous finding that urinary excretions of these plasma proteins selectively increased in parallel with enhanced glomerular filtration rate after acute protein loading, the present finding may be explained by renal hemodynamic changes, such as increased intraglomerular hydraulic pressure. PMID- 15111542 TI - Can we apply the National Cholesterol Education Program Adult Treatment Panel definition of the metabolic syndrome to Asians? AB - OBJECTIVE: Limited information is available about the metabolic syndrome in Asians. Furthermore, the definition of central obesity using waist circumference may not be appropriate for Asians. The objectives of this study were to determine the optimal waist circumference for diagnosing central obesity in Asians and to estimate the prevalence of the metabolic syndrome in an Asian population. RESEARCH DESIGN AND METHODS: We used data from the 1998 Singapore National Health Survey, a cross-sectional survey involving 4,723 men and women of Chinese, Malay, and Asian-Indian ethnicity aged 18-69 years. Receiver operating characteristic analysis suggested that waist circumference >80 cm in women and >90 cm in men was a more appropriate definition of central obesity in this population. The prevalence of the metabolic syndrome was then determined using the National Cholesterol Education Program Adult Treatment Panel III (NCEP ATP III) criteria with and without the modified waist circumference criteria. RESULTS: In Asians, decreasing waist circumference increased the crude prevalence of the metabolic syndrome from 12.2 to 17.9%. Using the modified Asian criteria, the prevalence of the metabolic syndrome increased from 2.9% in those aged 18-30 years to 31.0% in those aged 60-69 years. It was more common in men (prevalence 20.9% in men versus 15.5% in women; P < 0.001) and Asian Indians (prevalence 28.8% in Asian-Indians, 24.2% in Malays, and 14.8% in Chinese; P < 0.001). CONCLUSIONS: NCEP ATP III criteria, applied to an Asian population, will underestimate the population at risk. With a lower waist circumference cutoff, the prevalence of the metabolic syndrome is comparable to that in Western populations. Ethnic differences are likely to exist between populations across Asia. PMID- 15111543 TI - Does the association of habitual physical activity with the metabolic syndrome differ by level of cardiorespiratory fitness? AB - OBJECTIVE: Cardiovascular fitness (VO(2max)) and physical activity are both related to risk of metabolic disease. It is unclear, however, whether the metabolic effects of sedentary living are the same in fit and unfit individuals. The purpose of this study was, therefore, to describe the association between physical activity and the metabolic syndrome and to test whether fitness level modifies this relationship. RESEARCH DESIGN AND METHODS: Physical activity was measured objectively using individually calibrated heart rate against energy expenditure. VO(2max) was predicted from a submaximal exercise stress test. Fat mass and fat-free mass (FFM) were calculated using impedance biometry. A metabolic syndrome score was computed by summing the standardized values for obesity, hypertension, hyperglycemia, insulin resistance, hypertriglyceridemia, and the inverse level of HDL cholesterol and was expressed as a continuously distributed outcome. To correct for exposure measurement error, a random subsample (22% of cohort) re-attended for three repeat measurements in the year following the first assessment. RESULTS: The relationship of VO(2max) (ml O2.kg(FFM)(-1).min(-1)) and the metabolic syndrome score was of borderline significance after adjusting for age, sex, physical activity, and measurement error (beta = -0.58, P = 0.06). The magnitude of the association between physical activity (kJ.d(-1).kg(FFM)(-1)) and the metabolic syndrome was more than three times greater than for VO(2max) (standardized beta = -1.83, P = 0.0042). VO(2max), however, modified the relationship between physical activity energy expenditure and metabolic syndrome (P = 0.036). CONCLUSIONS: This study demonstrates a strong inverse association between physical activity and metabolic syndrome, an association that is much steeper in unfit individuals. Thus, prevention of metabolic disease may be most effective in the subset of unfit inactive people. PMID- 15111544 TI - Increasing incidence of diabetes after gestational diabetes: a long-term follow up in a Danish population. AB - OBJECTIVE: To study the incidence of diabetes among women with previous diet treated gestational diabetes mellitus (GDM) in the light of the general increasing incidence of overweight and diabetes and to identify risk factors for the development of diabetes. RESEARCH DESIGN AND METHODS: Women with diet-treated GDM during 1978-1985 (old cohort, n = 241, also followed up around 1990) or 1987 1996 (new cohort, n = 512) were examined in 2000-2002. Women were classified by a 2-h, 75-g oral glucose tolerance test according to the World Health Organization criteria or an intravenous glucagon test supplemented by measurement of GAD antibodies. Historical data from index-pregnancy and anthropometrical measurements were collected. RESULTS: A total of 481 (63.9%) women were examined (median 9.8 years [interquartile range 6.4-17.2]) after index pregnancy. Diabetes and impaired glucose tolerance (IGT)/impaired fasting glucose were present in 40.0 and 27.0% of women, respectively. In the new cohort, 40.9% had diabetes compared with 18.3% in the old cohort at the 1990 follow-up (P < 0.0005). Prepregnancy BMI was significantly higher in the new compared with the old cohort (26.0 [22.5-30.8] vs. 22.9 kg/m2 [20.2-28.0], P < 0.0005). Among others, new cohort membership, prepregnancy overweight (BMI > or = 25 kg/m2), and IGT postpartum were identified as independent predictors of diabetes by multiple logistic regression analyses. CONCLUSIONS: The incidence of diabetes among Danish women with previous diet-treated GDM was very high and had more than doubled over a 10-year period. This seems to be due to a substantial increase in BMI in women with GDM. PMID- 15111545 TI - HbA1c levels are significantly lower in early and late pregnancy. PMID- 15111546 TI - The case for intravenous arginine stimulation in lieu of mixed-meal tolerance tests as outcome measure for intervention studies in recent-onset type 1 diabetes. PMID- 15111547 TI - Heterophile antibodies masquerade as interferon-alpha in subjects with new-onset type 1 diabetes. PMID- 15111548 TI - Complete long-term recovery of beta-cell function in autoimmune type 1 diabetes after insulin treatment. PMID- 15111549 TI - Fasting and insulin glargine in individuals with type 1 diabetes. PMID- 15111550 TI - Body metabolism provides a foundation for noninvasive blood glucose monitoring. PMID- 15111551 TI - The fattening burden of type 2 diabetes on Mexicans: projections from early growth to adulthood. PMID- 15111552 TI - Ionized magnesium in Danish children with type 1 diabetes. PMID- 15111553 TI - A systematic review of adherence with medications for diabetes. AB - OBJECTIVE: The purpose of this study was to determine the extent to which patients omit doses of medications prescribed for diabetes. RESEARCH DESIGN AND METHODS: A literature search (1966-2003) was performed to identify reports with quantitative data on adherence with oral hypoglycemic agents (OHAs) and insulin and correlations between adherence rates and glycemic control. Adequate documentation of adherence was found in 15 retrospective studies of OHA prescription refill rates, 5 prospective electronic monitoring OHA studies, and 3 retrospective insulin studies. RESULTS: Retrospective analyses showed that adherence to OHA therapy ranged from 36 to 93% in patients remaining on treatment for 6-24 months. Prospective electronic monitoring studies documented that patients took 67-85% of OHA doses as prescribed. Electronic monitoring identified poor compliers for interventions that improved adherence (61-79%; P < 0.05). Young patients filled prescriptions for one-third of prescribed insulin doses. Insulin adherence among patients with type 2 diabetes was 62-64%. CONCLUSIONS: This review confirms that many patients for whom diabetes medication was prescribed were poor compliers with treatment, including both OHAs and insulin. However, electronic monitoring systems were useful in improving adherence for individual patients. Similar electronic monitoring systems for insulin administration could help healthcare providers determine patients needing additional support. PMID- 15111554 TI - Insulin storage in Europe: a comment to Grajower et al., Eli Lilly, and Novo Nordisk. PMID- 15111555 TI - Glycemic treatment: Control of glycemia. PMID- 15111556 TI - Difference in presentation of charcot osteoarthropathy in type 1 compared with type 2 diabetes. PMID- 15111557 TI - Status of research funding by the American Diabetes Association--5th and final year. PMID- 15111558 TI - Soluble CD40L in young type 1 diabetic individuals without clinical microvascular and macrovascular complications. PMID- 15111559 TI - Strong inverse correlation between serum adiponectin level and heart rate corrected QT interval in an apparently healthy population: a suggestion for a direct antiatherogenic effect of adiponectin. PMID- 15111560 TI - Seasonal changes in body composition and blood HbA1c levels without weight change in male patients with type 2 diabetes treated with insulin. PMID- 15111561 TI - Reversibile quadriplegia and nonketotic hyperosmolar coma: is it an exceptional association or an overlooked complication? PMID- 15111562 TI - Relationship between moderate alcohol consumption and adiponectin and insulin sensitivity in a large heterogeneous population. PMID- 15111563 TI - How long should insulin be used once a vial is started? PMID- 15111565 TI - The metabolic syndrome: the emperor needs some consistent clothes. PMID- 15111566 TI - Aspirin resistance in diabetic patients. PMID- 15111568 TI - Is aspirin resistance a real problem in people with type 2 diabetes? PMID- 15111569 TI - Autoimmune hypoglycemia in a type 2 diabetic patient with anti-insulin and insulin receptor antibodies. PMID- 15111572 TI - Diagnosing insulin resistance by simple quantitative methods in subjects with normal glucose metabolism. PMID- 15111573 TI - Benefits and risks of solitary islet transplantation for type 1 diabetes using steroid-sparing immunosuppression. PMID- 15111576 TI - Type 2 diabetes prevalence in Asian subjects. PMID- 15111578 TI - Modulation of Sub-RPE deposits in vitro: a potential model for age-related macular degeneration. AB - PURPOSE: Sub-RPE deposits form in a variety of conditions most notably in age related macular degeneration. The purpose of this study was to generate sub-RPE deposits in vitro and to test the hypotheses that high protein concentrations or retinal homogenate increase deposit formation and that a challenge with tumor necrosis factor (TNF)-alpha or metalloproteinase (MMP)-2 decreases such deposits. METHODS: ARPE-19 cells were grown on plastic and on collagen type I-coated membrane inserts in media containing various concentrations of fetal calf serum (FCS), bovine serum albumin, or porcine retinal homogenate. In addition, cells grown on membrane inserts were treated with TNF-alpha or MMP-2. Sub-RPE deposits were assessed by electron microscopy and classified into fibrillar, condensed, banded, and membranous subtypes. The area of the micrograph occupied by each type was estimated with a point-counting technique. MMP-2 activity was assessed in tissue culture supernatants by zymography. RESULTS: With increasing time in culture, total deposit formation did not change, but the amount of condensed material deposited by ARPE-19 cells increased while the fibrillar component decreased. Albumin challenge resulted in an increased amount of deposit, predominantly of the membranous type. Challenge with retinal homogenate led to a greater net deposit formation with significant increases in the condensed and banded forms. Cells treated with TNF-alpha or MMP-2 showed a dramatic reduction in all types of sub-RPE deposit. Zymography demonstrated that unchallenged cells produced predominantly MMP-2. Retinal homogenate challenge reduced the total amount of active MMP-2 produced, and TNF-alpha stimulated MMP-9 production. CONCLUSIONS: Sub-RPE deposits formed in vitro share ultrastructural features with those seen in vivo. Deposit formation can be modulated by challenge with retinal homogenate, TNF-alpha, or MMP-2. Significantly, the results provide proof of the principle that sub-RPE deposits can be formed and modified in vitro. PMID- 15111579 TI - Inhibition of ocular angiogenesis by an adenovirus carrying the human von Hippel Lindau tumor-suppressor gene in vivo. AB - PURPOSE: The purpose of this study was to investigate the effect of the von Hippel-Lindau (VHL) protein on VEGF gene expression in vitro and to determine whether adenovirus-mediated VHL intraocular gene transfer inhibits the development of angiogenesis in a monkey model of multiple branch retinal vein occlusion (BRVO). METHODS: A recombinant adenovirus vector adVHL was constructed to deliver the human VHL gene. Total RNA prepared from various kinds of cells transduced with adLacZ (control) or adVHL under normoxic or hypoxic conditions was subjected to Northern blot analyses. Either adLacZ or adVHL was delivered by preretinal injection in monkeys. The effects of adLacZ or adVHL on ocular neovascularization in laser-induced multiple BRVO was evaluated in color photographs and with fluorescein angiography (FA). RESULTS: VHL expression in adVHL-transduced cells was confirmed at the transcript and protein levels. VHL overexpression significantly decreased the levels of VEGF transcripts in human aortic endothelial cells (HAECs); retinal pigment epithelium (RPE) cells; and RCC 786-O cells, renal carcinoma cells lacking VHL expression under normoxia. In contrast, VHL had no effect on the hypoxia-mediated increase in VEGF expression in these cells, although basal levels of VEGF expression were substantially reduced. Color photographs and FA revealed that retinal neovascularization and iris rubeosis accompanied by multiple BRVO in a monkey model were obviously suppressed by VHL overexpression. Northern blot analysis and immunostaining for VHL and VEGF indicated that VHL transfer obviously suppressed VEGF gene expression in VHL-transduced tissues such as retina or RPE. CONCLUSIONS: The results showed that adenovirus expressing VHL led to a significant reduction in VEGF expression in vitro under normoxic or hypoxic conditions. adVHL effectively inhibited angiogenesis in retina and iris in laser-induced multiple BRVO in monkey eyes. These data suggest that gene therapy based on VHL gene delivery has potential in the treatment of human ocular neovascularization. PMID- 15111580 TI - Prosaposin gene expression in normal and dystrophic RCS rat retina. AB - PURPOSE: To identify proteins secreted by the retinal pigment epithelium (RPE) and to analyze their cellular distribution in normal and pathologic rat retinas at various stages of eye development. METHODS: A cDNA library was constructed with RNA isolated from porcine RPE sheets and screened by using the yeast signal sequence trap system. In situ hybridization, immunohistochemistry, and semiquantitative RT-PCR analysis were performed on rat retinas. RESULTS: The cDNA encoding prosaposin was isolated. This is the first time this gene has been shown to be expressed in the retina. Prosaposin mRNA was detected in the rat RPE cell monolayer and in ganglion cells 14, 21, and 45 days after birth. The amount of prosaposin mRNA increased between days 14 and 45 after birth in normal retinas (rdy+), but not in the pathologic retinas (rdy-) of RCS rats. CONCLUSIONS: Several techniques were used to determine the localization of prosaposin in rat retinas. The increase in the amount of prosaposin mRNA in normal retinas coincided with the maturation of photoreceptor cells and the beginning of the phagocytosis process. In addition, the RCS rdy- RPE cells, characterized by the abrogation of the ingestion phase of the photoreceptor outer segments, are deficient in prosaposin expression. PMID- 15111581 TI - Association of apolipoprotein E alleles with susceptibility to age-related macular degeneration in a large cohort from a single center. AB - PURPOSE: To examine the effect of apolipoprotein E (APOE) alleles on age-related macular degeneration (AMD) risk and on age at diagnosis of AMD in a large patient cohort recruited from a single center. METHODS: The frequency of APOE alleles was analyzed in 632 unrelated AMD patients and 206 unrelated controls, all of whom were of white ancestry. The presence or absence of disease symptoms in all patients and controls was based on clinical examination and/or ophthalmic records. The association with APOE was explored in the context of AMD subtypes, family history status, possible interaction with smoking, and distribution of age at diagnosis of AMD. RESULTS: The frequency of the epsilon4 allele was significantly reduced in patients compared with controls (0.10 vs. 0.14, P < or = 0.02). Gender- and age-adjusted odds ratios indicated that epsilon4-carriers have significantly lower risk of developing AMD compared to epsilon3epsilon3 subjects (OR = 0.55, 95% CI: 0.37-0.82, P = 0.004). In the cohort, AMD patients with a positive family history exhibited a significant 3.5 years earlier age at diagnosis (P = 0.001); however, APOE alleles did not appear to modulate the age at diagnosis of AMD. CONCLUSIONS: The association between the APOE-epsilon4 allele and a reduced risk of AMD was established in a large cohort with sufficient statistical power. How distinct APOE alleles affect AMD susceptibility warrants further investigation. PMID- 15111582 TI - The epsilon2 and epsilon4 alleles of the apolipoprotein gene are associated with age-related macular degeneration. AB - PURPOSE: To date, of all the genes studied in relation to age-related macular degeneration (AMD), the alleles of the apolipoprotein (apoE) gene have been the most consistently associated with disease. However, not all apoE studies have found an association, and among these the associations differ. The current study was conducted to investigate further the association of this gene in AMD. METHODS: Three hundred twenty-two unrelated individuals with diagnosed AMD and 123 unrelated but ethnically matched control subjects were analyzed. All subjects completed a standard questionnaire and were given a fundus examination. A blood sample was collected for DNA extraction. The common allelic variants of apoE were screened through the use of polymerase chain reaction (PCR) and restriction enzyme digestion followed by statistical analysis. RESULTS: Individuals with the epsilon3 epsilon4 genotype of apoE had an approximate halving of disease risk for late (end-stage) AMD (odds ratio [OR] 0.58, 95% confidence interval [CI] 0.34 0.98) relative to the epsilon3 epsilon3 genotype at age of ascertainment. Stratification of late AMD into atrophic and neovascular disease revealed that the greatest protective effect for the epsilon3 epsilon4 genotype was in individuals with atrophic disease (OR 0.35, 95% CI 0.13-0.92). Men with the epsilon3 epsilon4 genotype also showed almost a threefold reduction in risk of disease in late AMD (OR 0.36, 95% CI 0.16-0.82). However, individuals with late AMD and the epsilon2 epsilon3 genotype had a significantly earlier mean age of diagnosis of disease (3.4 years, P = 0.015) compared with those with the epsilon3 epsilon3 genotype, and this was most evident in women (3.9 years, P = 0.011) and in individuals with neovascular disease (4.7 years, P = 0.003). CONCLUSIONS: The alleles of apoE appear to have a role in the etiology of AMD, with the epsilon4 allele being protective, or at the very least, delaying the age of diagnosis of disease, whereas the epsilon2 allele appears to have a modifier effect by bringing forward the mean age of disease diagnosis. PMID- 15111583 TI - Incidence and cumulative risk of childhood cataract in a cohort of 2.6 million Danish children. AB - PURPOSE: To determine the incidence and cumulative risk of childhood cataract in Denmark during 1980 to 2000. METHODS: A cohort of 2,616,439 Danish children born between 1962 and 2000 was followed from 1980 or from the day of birth, whichever occurred later, until their 18th birthday, death, emigration, or diagnosis of cataract, whichever occurred first. Cases were ascertained from the Danish National Register of Patients (NRP) and validated by reviewing the medical records. They were divided into four groups: congenital/infantile (CI) cataract, traumatic cataract, complicated cataract, and "other" types of cataract. RESULTS: After diagnostic validation, 1311 children with cataract (59% with CI cataract) were included in the study. During 1995 to 2000 the overall cumulative risk of childhood cataract was 108.4 per 100,000 children. There was no significant difference in incidence between girls and boys or over time (1980 to 2000) for CI, complicated, and "other" types of cataract. In contrast, the incidence of traumatic cataract was significantly higher among boys. It remained increased during the entire study period despite a 23% decrease per 5 years among boys. Sixty-six percent of the children diagnosed with CI cataract below 2 years of age underwent surgery within 1 year. CONCLUSIONS: The stable incidence during a 20 year period of CI cataract and complicated cataract indicates that risk factors for these conditions have remained unchanged, whereas the marked drop of traumatic cataract among boys most likely reflects changed behavior and an increased focus on preventive measures. PMID- 15111584 TI - Significant nonsurgical risk factors for endophthalmitis after cataract surgery: EPSWA fourth report. AB - PURPOSE: Endophthalmitis remains a devastating complication of cataract surgery, despite improved methods of prophylaxis and surgical technique. The current study was conducted to identify sociodemographic, environmental, and clinical risk factors for the development of postoperative endophthalmitis, using population based administrative data from Western Australia. METHODS: The Western Australian Data Linkage System identified all patients who underwent cataract surgery, along with those in whom postoperative endophthalmitis subsequently developed, from 1980 to 2000 inclusive. Cases of endophthalmitis were cross-referenced with other sources and validated by medical record review. After selection and preliminary analysis of potential risk factors, multivariate logistic regression modeling was used to estimate odds ratios for the selected variables. RESULTS: Over the 21 years, 210 cases of endophthalmitis occurred after 117,083 cataract procedures, yielding a cumulative incidence rate of 1.79 per 1000 procedures. The incidence of endophthalmitis decreased for extracapsular extraction over the whole period, but not for phacoemulsification over the recent 12 years. There was no risk adjusted difference in the incidence rate of endophthalmitis for the various cataract surgery procedure types. However, a significantly higher risk was found in patients aged over 80 years, in having surgery in private hospitals, and to a lesser degree in having same-day surgery and surgery in winter. Cataract surgery with lacrimal or eyelid procedures dramatically increased the risk of endophthalmitis. CONCLUSIONS: It may be possible to reduce the incidence rate of postoperative endophthalmitis by almost 80% with a systematic approach to the management of elderly patients, hospital stay, and clinical protocols. PMID- 15111585 TI - Urokinase-type plasminogen activator to prevent haze after photorefractive keratectomy, and pregnancy as a risk factor for haze in rabbits. AB - PURPOSE: To observe the effect of urokinase-type plasminogen activator (uPA) on the development of subepithelial haze after photorefractive keratectomy (PRK) and to assess pregnancy as a risk factor for haze. METHODS: In 30 rabbits, both eyes of 27 were subjected to PRK and both eyes of 3 served as the nonsurgical control. In the first part of the experiment, for 7 days after PRK, three rabbits (one was pregnant) received aprotinin in one eye and no aprotinin in the contralateral eye. uPA activity was measured by a spectrophotometric method from tear samples in these eyes. In the second part, for 5 days after PRK, 24 rabbits (8 were pregnant) were treated with uPA in one eye and no uPA in the contralateral eye. Haze grading was performed according to the system of Hanna. RESULTS: In the first experiment, the aprotinin-treated eyes and the aprotinin-untreated eye of the pregnant rabbit showed development of haze. In the second, there were clear corneas in 24 of 24 uPA-treated eyes and in 15 of 24 uPA-untreated eyes. Post-PRK haze formed in 9 of 24 uPA-untreated eyes (7 of the 9 haze observations in pregnant rabbits). Within the uPA-untreated group, haze formed in corneas of 7 of 8 pregnant versus 2 of 16 nonpregnant rabbits. There was a strong association between the uPA treatment and clear corneas (P = 0.003) and between pregnancy and haze (P = 0.002). CONCLUSIONS: The present results suggest that pregnancy is a risk factor for post-PRK haze in untreated rabbit eyes and that uPA is effective in preventing the occurrence of haze. PMID- 15111586 TI - Corneal healing after uncomplicated LASIK and its relationship to refractive changes: a six-month prospective confocal study. AB - PURPOSE: To investigate corneal healing and the factor(s) possibly responsible for refractive changes after laser in situ keratomileusis (LASIK). METHODS: Twenty eyes of 10 patients who underwent LASIK for myopia were examined clinically and by real-time confocal microscopy for 6 months. Epithelial and posterior stromal thicknesses and the thickness of the keratocyte activation zone were measured, and refractive changes were compared with these values. Keratocyte morphology, flap thickness, and subbasal nerve fiber bundle morphology after LASIK were also investigated. RESULTS: No significant change was detected over time in epithelial thickness after LASIK treatment; however, the posterior stromal thickness was found to be significantly higher 1 month after surgery. A slight but statistically significant negative correlation was detected between the thickness of the keratocyte activation zone and the spheroequivalent refraction after LASIK. The subbasal nerve fiber bundle's morphology returned to its preoperative appearance 6 months after LASIK, but in the flap stroma the nerve fiber bundle morphology remained abnormal at 6 months after LASIK surgery. CONCLUSIONS: A weak but significant negative correlation between the thickness of the keratocyte activation zone and spheroequivalent refraction was found after LASIK. The different refractive properties of activated keratocytes may be responsible for the myopic shift after LASIK. Further studies are needed to clarify this hypothesis. PMID- 15111587 TI - Effect of overexpressing the transcription factor E2F2 on cell cycle progression in rabbit corneal endothelial cells. AB - PURPOSE: To test the hypothesis that overexpression of the transcription factor E2F2 promotes cell cycle progression in nonproliferating corneal endothelial cells. METHODS: Ex vivo rabbit corneas were transfected with a lipid transfection reagent and either a control plasmid containing full-length cDNA for enhanced green fluorescent protein (pIRES2-EGFP) or a plasmid containing full-length cDNA for both E2F2 and EGFP (pIRES2-E2F2/EGFP). Transfection control experiments consisted of corneas incubated in buffer without transfection reagent or plasmid or incubated in reagent without plasmid. After transfection, corneas were incubated for various periods in 0.1% FBS (a concentration that maintains cell health, but does not promote proliferation). Immunocytochemical (ICC) localization tested for overexpression of E2F2 in transfected corneal endothelial cells and permitted calculation of transfection efficiency. Endothelial cell viability was tested in transfected ex vivo corneas and confluent cultures by using a cell-viability assay. Apoptosis was detected in confluent cultures by TUNEL assay. RT-PCR tested for mRNA expression of Ki67 (a marker of actively cycling cells) and cyclin B1 (a marker for the G2-phase of the cell cycle). Semiquantitative densitometric analysis compared the relative amounts of PCR reaction products. RESULTS: ICC demonstrated the colocalization of E2F2 and EGFP in corneal endothelium with a transfection efficiency of 10% to 12%, using the pIRES2-based plasmid and transfection reagent. The cell-viability assay revealed very few dead cells in ex vivo corneal endothelium that overexpressed E2F2. Cell viability and TUNEL assays of confluent cultures revealed that approximately 27% of cells died in all cultures incubated with transfection reagent, but death appeared not to be due to apoptosis. No additional cell death was noted by either assay in cells that overexpressed E2F2. RT-PCR of endothelial samples obtained 48 hours after transfection showed the presence of higher levels of reaction product for Ki67 (a 5.1-fold increase) and cyclin B1 (a 2.3-fold increase) in cells that overexpressed E2F2 than in control samples. CONCLUSIONS: Overexpression of the transcription factor E2F2 in nonproliferating rabbit corneal endothelial cells induces cell cycle progression without inducing significant apoptosis. PMID- 15111588 TI - Corneal asphericity change after excimer laser hyperopic surgery: theoretical effects on corneal profiles and corresponding Zernike expansions. AB - PURPOSE: To determine the theoretical relationships between the changes in corneal paraxial power, asphericity, and the corresponding Zernike polynomial expansion after conventional and customized excimer laser correction of hyperopia. METHODS: The initial corneal profile was modeled as a conic section of apical radius of curvature R1 and asphericity Q1. The theoretical value of the postoperative apical radius of curvature R2 was computed by using a paraxial formula from the value of R1 and hyperopic defocus D. The postoperative asphericity Q2 of the corneal surface was computed within the optical zone of diameter S after the delivery of a Munnerlyn-based profile of ablation for hyperopia using conic section-fitting and minimization of the squared residuals. These calculations were repeated for different values of defocus, initial apical radius of curvature, and asphericity. Taylor series expansions were also used to provide an approximation aimed at predicting change in asphericity. The coefficients of a Zernike polynomial expansion of the rotationally symmetrical corneal profile (defocus C2(0), spherical aberration C4(0), secondary spherical aberration C6(0)) were also computed, by using scalar products applied to the considered corneal profile modeled as a conic section and were expressed as a function of both its apical radius and asphericity. This allowed approximation of the variations of the Zernike polynomial expansion of the corneal profiles by subtracting the postoperative coefficient weighting a particular aberration from that of the preoperative one in different theoretical situations, after both conventional and customized hyperopia treatments aimed at controlling the postoperative corneal asphericity and delivered over a normalized pupil diameter. RESULTS: Conical least-squares fitting was unambiguous, allowing approximation of the postoperative corneal profile as a conic section of apical radius R2. After a Munnerlyn-based hyperopia treatment, the sign of the asphericity of this profile remains theoretically unchanged, but its value decreased for initially oblate and increased for initially prolate corneas, respectively. A similar trend was noted with the approximation obtained by the Taylor series expansion. The alteration of the apical radius and/or of the asphericity of the corneal surface resulted in variations of both the corneal profile Zernike coefficients C2(0) and C4(0). The former was essentially dependent on the variation of the apical radius and the latter essentially on the variation of both apical radius and asphericity. CONCLUSIONS: Conventional and customized profiles of ablation for hyperopia alter the postoperative corneal asphericity and the Zernike coefficients of the corneal profile. The results of this study may be useful in the interpretation of the postoperative variations of the corneal profile and their impact on corneal wavefront expansion variations after both conventional and customized profiles of ablation. PMID- 15111589 TI - Conjunctival proinflammatory and proapoptotic effects of latanoprost and preserved and unpreserved timolol: an ex vivo and in vitro study. AB - PURPOSE: To compare the toxicity of latanoprost and preserved and unpreserved timolol on conjunctival cells. Expression of inflammatory markers and MUC5AC related mucin production were evaluated by impression cytology in a case-control ex vivo study. The proapoptotic effect of the same drugs was also evaluated in vitro in a conjunctival cell line and compared with that of benzalkonium chloride (BAC). METHODS: Impression cytology (IC) specimens were obtained from a series of normal subjects and from patients with glaucoma treated for at least 1 year with latanoprost eye drops or preserved or unpreserved timolol. All groups were comparable in age and duration of treatment. Expression of HLA-DR, intercellular adhesion molecule (ICAM)-1, and mucin was evaluated in a masked manner by flow cytometry. For the in vitro study, a human conjunctiva-derived cell line was treated with 0.02% BAC-containing latanoprost or timolol, unpreserved timolol, or 0.02% BAC alone for 15 minutes, followed or not by 4 or 24 hours of cell recovery in normal medium. Cell viability and chromatin condensation were evaluated using microplate cold light cytofluorometry with the neutral red and the Hoechst 33342 tests, respectively. The Hoechst-neutral red ratio was defined for the apoptosis assay, and cytoskeleton changes were assessed by confocal microscopy. RESULTS: No difference was found between normal eyes and those receiving unpreserved timolol. Preserved latanoprost and timolol significantly increased the inflammatory marker expression and decreased MUC5AC expression, but to a significantly higher extent in the preserved timolol group compared with latanoprost. In vitro, 0.02% BAC containing timolol and latanoprost triggered conjunctival cell apoptosis-however, to a significantly lesser extent than did 0.02% BAC alone. Unpreserved timolol did not cause any cell toxicity. CONCLUSIONS: These ex vivo and in vitro studies demonstrate that BAC-containing latanoprost and timolol exhibit higher proinflammatory and proapoptotic effects on conjunctival cells than does unpreserved timolol. Latanoprost caused less toxicity, however, than preserved timolol, and both drugs were less toxic than BAC alone. These results suggest a potential protective effect of the prostaglandin analogue and to a lesser extent of timolol against the toxicity of BAC in conjunctival cells. PMID- 15111590 TI - A new noninvasive tear stability analysis system for the assessment of dry eyes. AB - PURPOSE: This was a prospective case-control study conducted to evaluate the effectiveness of the Tear Stability Analysis System (TSAS) for the assessment of patients with dry eye. METHODS: The TSAS can take 10 consecutive corneal topograms at one per second for 10 seconds. Examinations using the TSAS were conducted in 26 eyes of 26 healthy control subjects and in 27 eyes of 27 patients with dry eye. Examinations were also conducted in 14 eyes of 14 patients before and after the insertion of punctum plugs. Surface regularity and asymmetry indices (SRI, SAI), as well as new tear stability regularity and asymmetry indices (TSRI, TSAI), derived from SRI and SAI, were analyzed. RESULTS: The mean SRI and SAI in dry eyes were significantly greater than in control eyes (P < 0.05). The time-wise change of SRI and SAI was significantly different between dry eyes and control eyes (P < 0.05). TSRI and TSAI in dry eyes were also significantly greater than in control eyes. Punctum plug insertion was associated with a significant decrease in SRI and SAI (P < 0.05). CONCLUSIONS: TSAS was effective in objectively assessing the tear stability in patients with dry eye. This system may be useful in noninvasive diagnosis of dry eye and evaluation of treatment effects. PMID- 15111591 TI - Prophylactic effect of IL-10 gene transfer on induced autoimmune dacryoadenitis. AB - PURPOSE: To evaluate the effect of viral IL-10 on the lacrimal gland immunopathologic response in the ocular surface disease, induced autoimmune dacryoadenitis. METHODS: Disease was induced in rabbits by injecting inferior lacrimal glands with peripheral blood lymphocytes activated by 5 days of coculture with autologous acinar cells in a mixed-cell reaction. In the treated group, an adenoviral vector carrying the vIL-10 gene was concurrently injected with activated lymphocytes. Tears were collected periodically for quantitation of IL-10 by ELISA. Two weeks after disease induction, tear production, tear film breakup time, and rose bengal staining score were determined. Sectioned glands were immunostained for expression of CD4, CD8, rabbit thymic lymphocyte antigen (RTLA), CD18 and major histocompatibility complex class II. RESULTS: The titer of vIL-10 in tears was at its maximum on day 3, started to decline by day 7, and was undetectable by day 14. In the diseased group, the tear production rate and tear film breakup time were significantly decreased, and rose bengal staining was significantly increased. Diseased glands had immune cell infiltrates containing CD4+, RTLA+, and CD18+ cells, and major histocompatibility complex class II expression was increased. These changes were significantly ameliorated by expression of vIL-10. CONCLUSIONS: In vivo transduction of the lacrimal gland with AdvIL-10 resulted in the transient appearance of vIL-10 in tears. The presence of vIL-10 partially suppressed the appearance of Sjogren-syndrome-like features of reduced tear production, accelerated tear breakup, ocular surface disease, and immunopathologic response. Anti-inflammatory cytokine gene expression may offer a therapeutic modality for the treatment of autoimmune dacryoadenitis, once suitable vectors become available. PMID- 15111592 TI - Two mutations in the TGFBI (BIGH3) gene associated with lattice corneal dystrophy in an extensively studied family. AB - PURPOSE: To determine the genetic basis for lattice corneal dystrophy (LCD) in an extensively studied family. METHODS: Ten affected family members were examined clinically, and three individuals were studied with in vivo confocal microscopy and optical coherence tomography (OCT). Corneal tissues from eight affected family members were examined histopathologically. The status of the transforming growth factor beta-induced gene (TGFBI) gene was determined in each consenting family member (six affected, seven nonaffected) by amplifying, sequencing, and analyzing exons 4 and 12 of TGFBI for mutations. All exons from the entire coding region of TGFBI of one affected person were analyzed for mutations. RESULTS: Slit lamp biomicroscopy disclosed the clinical features of LCD in both eyes of affected individuals. In vivo confocal microscopy confirmed the presence of deposits as bright lesions within the corneal stroma. OCT revealed increased reflectivity within the corneal stroma. The corneal stroma in persons undergoing penetrating keratoplasty contained amyloid. Affected members of the family were found to have two heterozygous single-nucleotide mutations in exon 12 of the TGFBI gene (C1637A and C1652A) leading to predicted amino acid substitutions in the encoded TGFbeta-induced protein (A546D and P551Q). Mutations were not detected in exon 4. In addition, an inconsequential single-nucleotide polymorphism T1620C (F540F) was found in some affected and nonaffected family members. CONCLUSIONS: Two mutations in the TGFBI gene (A546D and P551Q) cosegregated with LCD in an extensively studied family that lacked the R124C mutation that frequently accompanies this form of corneal amyloidosis. PMID- 15111593 TI - Expression analysis of the matrix GLA protein and VE-cadherin gene promoters in the outflow pathway. AB - PURPOSE: To test the ability of promoter fragments from the matrix Gla protein (MGP) and vascular endothelial-cadherin (VE-cad) genes to target gene expression in a specific manner in the cells of the outflow pathway, by using adenoviral mediated gene transfer in organ culture. METHODS: Perfused anterior segments of human eyes were infected with replication-deficient recombinant adenoviruses expressing the beta-galactosidase reporter gene driven by the cytomegalovirus (CMV; control, n = 6), MGP (n = 6), or VE-cad (n = 12) promoters. Forty-eight hours after infection, the anterior segments were fixed and stained for beta galactosidase activity. The distribution of beta-galactosidase expression was analyzed in paraffin-embedded sections. RESULTS: The MGP promoter fragment resulted in beta-galactosidase expression by the cells of the conventional outflow pathway and did not show any activity in the corneal endothelium or other cells posterior to the scleral spur. Adenovirus containing the VE-cad promoter fragment showed functionality of the promoter in vascular endothelial cells, but failed to produce any detectable expression in the cells of the outflow pathway. CONCLUSIONS: Directed expression by the MGP gene promoter specifically to the trabecular meshwork (TM) provides a new tool for specific gene transfer to the outflow pathway. Results with the VE-cad promoter fragment indicate possible differences in the regulation of this gene between vascular and Schlemm's canal endothelial cells. Taken together, these data demonstrate the feasibility of targeted gene expression to the outflow pathway cells using tissue specific promoters. PMID- 15111594 TI - Optimizing and validating an approach for identifying glaucomatous change in optic nerve topography. AB - PURPOSE: To determine and validate optimal parameters for analysis in a previously described approach for identifying glaucomatous optic nerve progression by scanning laser tomography. METHODS: Thirty-degree sectors of rim area, as defined by an experimental reference plane, were analyzed for change with respect to different statistical limits of variability (80%, 90%, 95%, 98%, 99%, and 99.9%) in the longitudinal image series of 62 eyes from 30 ocular hypertension converters and 32 normal control subjects. A criterion requiring that change is repeatable in two of three consecutive tests (the 2-of-3 criterion) was compared with a single-test strategy not requiring confirmation, and four other plausible criteria. The influence of these various parameters on sensitivity and the false-positive rate was evaluated. The same series were also assessed for change by the known method of computer-generated probability maps. RESULTS: More sectors were identified as progressing in converter eyes than in control eyes at every limit of variability. With stricter limits of variability and a requirement of confirmation, fewer sectors were identified as changing, especially in control eyes. The 2-of-3 criterion had the most favorably balanced sensitivity and false-positive rates: these were, for the 90% limit of variability, 90.0% and 6.2%, respectively, and for the 95% limit, 83.3% and 3.1%, respectively. Confirmed rim loss in converter eyes was most frequent in the disc poles and corresponded with the field hemisphere of conversion in 80%. Probability maps detected significant and repeatable change in 26 (86.7%) of 30 converter eyes and 14 (43.8%) of 32 of control eyes. CONCLUSIONS: This study was conducted to optimize and validate an approach for identifying progression. The method distinguished eyes with glaucomatous change from unchanging control eyes. PMID- 15111595 TI - Quantifying retinal nerve fiber layer thickness histologically: a novel approach to sectioning of the retina. AB - PURPOSE: To present a technique of ocular sectioning that enables continuous histologic measurements of peripapillary retinal nerve fiber layer (RNFL) thickness in a concentric ring around the optic disc, corresponding to similar regions measured by in vivo imaging techniques. METHODS: Two pig eyes and two normal human eyes were processed using the "umbrella" technique, in which peripapillary concentric ring sections were obtained, at increasing diameters, all centered on the optic disc. Each histologic ring section contains a continuous circumferential 360 degrees retinal slice, oriented approximately perpendicular to the retinal surface. Every histologic slice contains each axon of the retina, sectioned perpendicular to each axon's long axis and at an equal set distance from the disc margin. RESULTS: Ring sections from pig and human eyes are presented and correlated to known RNFL anatomy. For the two human eyes, peripapillary RNFL thickness was quantified and plotted, resulting in the expected double-hump pattern. CONCLUSIONS: The umbrella technique provides, on a single histologic section, all necessary information for quantifying the entire RNFL layer of that eye, in a standardized fashion. This technique can simplify the process of analyzing the RNFL thickness histologically, assist in obtaining a normative database of RNFL thickness in humans, and be implemented as a histologic end point in animal studies evaluating new treatment modalities for glaucoma. PMID- 15111597 TI - Laboratory investigation of Acanthamoeba lugdunensis from patients with keratitis. AB - PURPOSE: Species of four Acanthamoeba isolates (KA/E2, KA/E12, KA/E15, and KA/E16) from the cornea of patients with keratitis were identified and their molecular characteristics compared with those of other strains. METHODS: Morphologic features of amebic cysts were evaluated with a microscope with differential interference contrast (DIC) optics. Restriction fragment length polymorphisms (RFLP) of mitochondrial DNA (mtDNA), riboprinting of small subunit ribosomal RNA gene (18S rDNA), and DNA sequences of 18S rDNA were analyzed. mtDNA and PCR-amplified 18S rDNA of the ocular isolates were digested with restriction enzymes, and the restriction patterns were compared with those of reference strains purchased from American Type Culture Collection (ATCC, Manassas, VA). PCR products of 18S rDNA were cloned and subjected to sequencing. The complete sequence of approximately 2300 bp obtained from the isolates and reference strains were compared with each other and those registered in GenBank. RESULTS: Three ocular isolates (KA/E2, KA/E12, and KA/E16) of Acanthamoeba revealed the identical mtDNA RFLPs and riboprint patterns with Acanthamoeba L3a, the type strain of A. lugdunensis. The other isolate (KA/E15) had riboprint patterns very similar to A. lugdunensis L3a but quite different mtDNA RFLP patterns from those of all the other strains. A dendrogram based on the riboprint data showed that three ocular isolates were identified as A. lugdunensis and the other isolate was very closely related to this species. Identification of the isolates as A. lugdunensis was confirmed by 18S rDNA sequence analysis. The sequence differences of the four isolates from A. lugdunensis L3a was 0.1% to 0.4% (3 to 8/2284 bp) and 1.2% to 1.5% from A. castellanii Castellani. CONCLUSIONS: This is the first report of Acanthamoeba keratitis in Korea caused by A. lugdunensis, which was originally isolated from a freshwater pool in France. Riboprinting can be used as a simple and rapid tool for putative identification of unknown Acanthamoeba ocular isolates. PMID- 15111596 TI - Increased expression of iron-regulating genes in monkey and human glaucoma. AB - PURPOSE: To understand the mechanisms mediating retinal ganglion cell loss in glaucoma, the gene expression patterns were compared for transferrin, ceruloplasmin, and ferritin between normal and glaucomatous retina in monkey and human eyes. METHODS: Laser photocoagulation was used to produce unilateral experimental glaucoma in monkeys. Gene expression was assessed by in situ hybridization and quantitative reverse transcription polymerase chain reaction (PCR). Immunohistochemistry was used to examine the retinal expression of iron related proteins in the retina in experimental monkey glaucoma and human glaucoma. RESULTS: Comparison of glaucomatous with control monkey retinas demonstrated increased mRNA expression of transferrin, ceruloplasmin, and ferritin heavy and light chains. In situ hybridization localized retinal gene expression of transferrin mainly to the inner nuclear layer and ferritin to both the inner and outer nuclear layers. Immunohistochemical examination of monkey and human glaucoma for these iron-related proteins demonstrated increases at the protein level. CONCLUSIONS: Increased mRNA and protein levels of the iron regulating proteins transferrin, ceruloplasmin, and ferritin are present in glaucoma. Together, these results suggest the involvement of iron and copper metabolism and associated antioxidant systems in the pathogenesis of glaucoma. PMID- 15111598 TI - Regional differences in tyrosine kinase receptor signaling components determine differential growth patterns in the human lens. AB - PURPOSE: The lens epithelium can be separated into two regions, the nondividing central zone and the equator, the site of all division in the normal lens. In the present study, the distribution of epithelial growth factor (EGF)/epithelial growth factor receptor (EGFR) signaling components was investigated and related to mitotic distribution in the lens. METHODS: Anterior and equatorial regions of the native epithelium were prepared separately from donor lenses. In vitro capsular bags were prepared from donor eyes and cultured. Receptor distribution was determined by immunocytochemistry and RT-PCR. Western blot analysis of phospholipase C (PLC)-gamma and extracellular signal-regulated kinase (ERK; total and active) was performed on cell lysates. Function was determined by calcium imaging of Fura-2-AM-loaded cells and also, in the case of capsular bags, by cell growth. RESULTS: Immunocytochemistry and RT-PCR showed an even distribution of EGFR across the native epithelium. Whole lenses, however, exhibited only a calcium response to EGF (10 ng/mL) at the equatorial region. Western blot analysis demonstrated significantly greater expression of PLCgamma and ERK (total and active) in the equator than in the central region. Addition of EGF increased growth rates of cells in capsular bags and an EGFR inhibitor decreased rates. EGF also induced a calcium response in posterior capsule cells in the capsular bags. CONCLUSIONS: EGFR is evenly distributed across the entire epithelium, whereas related calcium signaling and expression of PLCgamma and ERK have a marked bias to the equator. Therefore, levels of downstream enzyme components rather than changes in receptor expression dictate EGFR signaling output in the normal lens. In the wounded lens (capsular bag) EGFR signaling persists in cells growing on the posterior capsule. PMID- 15111599 TI - CRYBA3/A1 gene mutation associated with suture-sparing autosomal dominant congenital nuclear cataract: a novel phenotype. AB - PURPOSE: To identify the genetic defect leading to the congenital nuclear cataract affecting a large five-generation Swiss family. METHODS: Family history and clinical data were recorded. The phenotype was documented by both slit lamp and Scheimpflug photography. One cortical lens was evaluated by electron microscopy after cataract extraction. Lenticular phenotyping and genotyping were performed independently with short tandem repeat polymorphism. Linkage analysis was performed, and candidate genes were PCR amplified and screened for mutations on both strands using direct sequencing. RESULTS: Affected individuals had a congenital nuclear lactescent cataract in both eyes. Linkage was observed on chromosome 17 for DNA marker D17S1857 (lod score: 3.44 at theta = 0). Direct sequencing of CRYBA3/A1, which maps to the vicinity, revealed an in-frame 3-bp deletion in exon 4 (279delGAG). This mutation involved a deletion of glycine-91, cosegregated in all affected individuals, and was not observed in unaffected individuals or in 250 normal control subjects from the same ethnic background. Electron microscopy showed that cortical lens fiber morphology was normal. CONCLUSIONS: The DeltaG91 mutation in CRYBA3/A1 is associated with an autosomal dominant congenital nuclear lactescent cataract. A splice mutation (IVS3+1G/A) in this gene has been reported in a zonular cataract with sutural opacities. These results indicate phenotypic heterogeneity related to mutations in this gene. PMID- 15111600 TI - Retinal colocalization and in vitro interaction of the glutamate transporter EAAT3 and the serum- and glucocorticoid-inducible kinase SGK1 [correction]. AB - PURPOSE: The serum- and glucocorticoid-inducible kinase SGK1 regulates several epithelial channels and transporters, the related protein kinase B (PKB) regulates glucose transport. SGK1 is expressed in the brain and could thus regulate glial and/or neuronal transport processes. The present study explores whether SGK1 is expressed in the retina and whether it regulates EAAT3, a Na(+) coupled glutamate transporter. EAAT3 is expressed in retinal ganglion cells and accomplishes the clearance of glutamate from synaptic clefts. METHODS: Immunohistochemistry was performed to test for retinal SGK1 expression. For functional analysis, cRNA encoding EAAT3 was injected into Xenopus oocytes with or without additional injection of wild-type SGK1, constitutively active (S422D)SGK1, inactive (K127N)SGK1, and/or constitutively active (T308D,S473D)PKB. Glutamate induced current (I(GLU)) was taken as a measure for transport. RESULTS: SGK1 is indeed expressed in several retinal cells including retinal ganglion cells where it is colocalized with EAAT3. In EAAT3-expressing Xenopus oocytes, glutamate-induced current was stimulated by coexpression of wild-type SGK1, constitutively active (S422D)SGK1, and constitutively active (T308D,S473D)PKB, but not by inactive (K127N)SGK1. CONCLUSIONS: SGK1 and EAAT3 are coexpressed in retinal neurons, and SGK1 serves to stimulate EAAT3. This function is shared by protein kinase B (PKB). The experiments reveal a novel mechanism regulating EAAT3, which may be essential for the function of the retinal ganglion cells. PMID- 15111601 TI - Comparison of objective retinal thickness analysis and subjective stereo fundus photography in diabetic macular edema. AB - PURPOSE: To compare assessment of retinal thickening by stereo fundus photographs and by Retinal Thickness Analysis (RTA). METHODS: Forty-degree stereo fundus photographs of the macular field, and RTA scans in 99 eyes of 53 diabetic patients were compared in both location and area of retinal thickening. The deviation in retinal thickness on RTA from the mean retinal thickness +/-2 SD in healthy controls was mapped in a spreadsheet and printed, assuming the same size as the grid with the subjectively assessed retinal thickening. Location was evaluated as retinal thickening being present or absent in nine subfields. Area was one of four categories: No retinal thickening, <1 disc area (DA), <2 DA, or <3 DA. STATISTICS: kappa-statistic. RESULTS: Exact agreement on location was found in 632 of 891 observations (70.9%). kappa = 0.17 (95% CI: 0.08-0.25). Pooling observations for subfields 2 to 5 (inner circle), and 6 to 9 (outer circle) showed exact agreement = 75.8% and kappa = 0.10 (95% CI: -0.06-0.25) and exact agreement = 65.2% and kappa = 0.24 (95% CI: 0.13-0.34), respectively. Twelve eyes, in which both methods assessed the same amount of retinal thickening, showed no agreement on location, and were compared only on location of retinal thickening. Exact agreement on area was found in 44 of 87 (50.6%) eyes. Weighted Kappa = 0.34 (95% CI: 0.23-0.45). CONCLUSIONS: The degree of agreement between subjectively (with stereo fundus photographs) and objectively (with RTA) assessed location and area of retinal thickening was poor, and fair, respectively. PMID- 15111602 TI - MERTK arginine-844-cysteine in a patient with severe rod-cone dystrophy: loss of mutant protein function in transfected cells. AB - PURPOSE: Mutations in the MERTK gene are responsible for retinal degeneration in the Royal College of Surgeons (RCS) rat and are a cause of human autosomal recessive retinitis pigmentosa (RP). This study reports the identification and functional analysis of novel MERTK mutations to provide information regarding whether they are causative of severe rod-cone degeneration in a young patient. METHODS: MERTK missense variants identified by single-strand conformational polymorphism (SSCP) and sequence analysis were introduced into expression constructs and used to transfect HEK293T cells. Recombinant protein expression was assayed with anti-MERTK and anti-phosphotyrosine antibodies. Protein turnover was assayed in pulse-chase studies of 35S-methionine incorporation. Transcript levels were determined by quantitative RT-PCR. RESULTS: Three MERTK sequence variants were identified in a patient with rod-cone dystrophy: R722X in exon 16 and R865W in exon 19 on the paternal allele and R844C in exon 19 on the maternal allele. The R844C sequence change affects an evolutionarily conserved amino acid residue and was not detected in unaffected individuals. In transfected HEK293Tcells, wild-type (wt) and W865 MERTK were expressed at equivalent levels and present in the plasma membrane, stimulated tyrosine phosphorylation, and induced significant rounding of the cell bodies. In contrast, C844 MERTK was expressed at low levels and did not stimulate tyrosine phosphorylation. In addition, the relative stability of C844 MERTK was significantly less than wt in assays of protein turnover. At age 13, the patient had 20/60 and 20/200 acuities, tunnel vision of 5 degrees centrally, and a far temporal peripheral crescent bilaterally, and ERGs were nondetectable. The fundi showed bull's-eye macular atrophy and widespread RPE thinning. CONCLUSIONS: The present study reports the identification of R844C, the first putative pathogenic MERTK missense mutation that results in severe retinal degeneration with childhood onset when in compound heterozygous form with a R722X allele. The loss of function of C844 MERTK is probably due to decreased protein stability. PMID- 15111603 TI - Hyperspectral imaging for measurement of oxygen saturation in the optic nerve head. AB - PURPOSE: To evaluate a hyperspectral imaging technique for monitoring relative spatial changes in retinal oxygen saturation. METHODS: The optic nerve head (ONH) and overlying vessels in cynomolgus monkey eyes were imaged with a fundus camera attached to a hyperspectral imaging system. Images were acquired with inspiration of room air and pure oxygen and at controlled intraocular pressures (IOP) of 15 mm Hg (normal) and 60 mm Hg (sustained for up to 5 minutes). Changes in relative blood oxygen saturation in the vessels and ONH were assessed from reflectance spectra. Saturation maps were derived from contributions of oxygenated and deoxygenated hemoglobin spectral signatures extracted from hyperspectral images. The results obtained with hyperspectral imaging were compared with known experimental outcomes. RESULTS: Pure oxygen markedly increased oxygen saturation in veins. Increases in arteries and the ONH were smaller. The results obtained with hyperspectral image analysis agreed with known changes in oxygen saturation from breathing experiments. Raising IOP reduced saturation in all structures and resulted in profound desaturation of arteries. During sustained high IOP, a rebound in saturation was observed in the ONH. Spatial maps clearly showed the saturation changes in arteries, veins, and surrounding tissues. CONCLUSIONS: Hyperspectral imaging can be adapted to measure and map relative oxygen saturation in retinal structures and the ONH in nonhuman primate eyes. PMID- 15111604 TI - Apoptosis in proliferative vitreoretinopathy. AB - PURPOSE: To study the involvement of apoptosis using different apoptosis markers in PVR pathogenesis. METHODS: The presence of mRNA coding for Fas, Fas ligand (FasL), and TNF-related apoptosis inducing ligand (TRAIL) was investigated in vitreous samples from 46 consecutive patients-25 with PVR, 11 with retinal detachment (RD) not complicated by PVR, and 10 with macular hole (MH)-using RT PCR. From previously examined vitreous samples, 21 PVR, 9 RD, and 10 MH were examined for their levels of TGF-beta2 protein with sandwich ELISA kits. Five epiretinal membranes excised from five patients with PVR were also examined for apoptotic cell death using the terminal deoxytransferase (TdT) mediated dUTP biotin nick end labeling (TUNEL) technique. RESULTS: FAS mRNA was detected in 72% of patients with PVR, 55% of patients with RD and 20% of patients with MH. TRAIL mRNA was detected in 67% of patients with PVR, 89% of patients with RD, and 20% of patients with MH. FasL mRNA was detected in 20% of patients with PVR, 9% of patients with RD, and 10% of patients with MH. The median levels of Fas and TRAIL mRNA were significantly higher (P < 0.05) in patients with PVR than in those with MH hole but between patients with PVR and those with RD the difference was not significant (P > 0.05). A significant difference was detected between RD and MH for TRAIL mRNA levels (P = 0.008). For FasL, no significant difference between groups was found. TGF-beta2 was detected in all investigated vitreous samples. A significant difference was found between the PVR and MH groups (P = 0.001) and between the RD and MH groups (P = 0.004), but not between the PVR and RD groups (P < 0.05). The level of TGF-beta2 was significantly correlated to the level of TRAIL mRNA (r = 0.86), but no correlation was found between TGF-beta2 and Fas mRNA levels (r = 0.21). Four of five examined PVR epiretinal membranes showed positive staining for apoptotic cells using the TUNEL technique. CONCLUSIONS: Apoptosis is one of the mechanisms that is involved in PVR pathogenesis. Different apoptosis markers suggest different pathways occur in PVR, including Fas/FasL, TRAIL, and TGF-beta2 mediated processes. PMID- 15111605 TI - Novel complex GUCY2D mutation in Japanese family with cone-rod dystrophy. AB - PURPOSE: All mutations in the retinal guanylate cyclase gene (GUCY2D) that causes autosomal dominant cone-rod dystrophy (CORD) are associated with an amino acid substitution in codon 838. A novel heterozygous complex missense mutation of I915T and G917R in the GUCY2D gene was found in a Japanese family with autosomal dominant CORD. The clinical features associated with this mutation were described. METHODS: Blood samples were collected from 27 patients with cone-rod or cone dystrophies and from 11 patients with macular dystrophy. Genomic DNA was extracted from peripheral leukocytes. All 18 coding exons of the GUCY2D gene were directly sequenced. The PCR product carrying a novel mutation was subcloned, and each allele was sequenced. A complete ophthalmologic examination was performed in members of the family with the novel mutation. RESULTS: A novel heterozygous complex missense mutation of T2817C and G2822C that would predict I915T and G917R amino acid substitutions, respectively, was found in an autosomal dominant CORD family. The two nucleotide changes were located on the same allele, and segregated with the disease. Two other known missense mutations of R838H and R838C were found in two other CORD families. The clinical phenotype associated with the novel mutation was similar to that with the Arg838 mutations. CONCLUSIONS: A heterozygous complex mutation of I915T and G917R in the GUCY2D gene caused autosomal dominant CORD, indicating that a heterozygous mutation that does not include a codon 838 substitution can lead to this ocular phenotype. PMID- 15111606 TI - Age, blood pressure, and vessel diameter as factors influencing the arterial retinal flicker response. AB - PURPOSE: The present study investigated whether age, blood pressure (BP), and baseline vessel diameter influence the retinal arterial response to flicker light. METHOD: Thirty healthy subjects (mean age, 46.3; range, 22-73 years) and 15 patients with untreated essential arterial hypertension (mean, 50.9; range, 26 69 years) were examined. The diameter of the retinal arterioles was measured by a Retinal Vessel Analyzer (RVA; Imedos, Weimar, Germany). Each examination consisted of a 100-second baseline measurement and five 20-second periods of flicker stimulation, followed by an 80-second observation period. The five stimulation periods were then averaged. The rectangular luminance flicker operated at 12.5 Hz at a wavelength of 530 to 600 nm. The baseline-corrected flicker response (bFR) was defined as the difference between the peak dilatation and subsequent constriction after flicker stimulation minus the fluctuation of the baseline. The BP was measured at 1-minute intervals during the examination. RESULTS: In 26 subjects with normal BP, flicker light induced a bFR of +6.4% +/- 2.7%. The bFR decreased nonsignificantly in healthy subjects with increasing age (y = 8.48-0.048x; r = 0.26). The baseline diameter did not influence the amplitude of the flicker response over a range of 70 to 140 measuring units. The hypertensive patients reacted with a bFR of +2.2% +/- 2.5% (P < 0.001). Four hitherto healthy subjects with elevated BP during the examination were excluded from analysis. CONCLUSIONS: A significant correlation of age and bFR was not found in the small sample examined. Untreated arterial hypertension appeared to be associated with a reduced flicker response. The value of such functional vessel properties in the screening of vasosclerosis and in diagnostics in arterial hypertension should be examined in further studies. PMID- 15111607 TI - The influence of path length and matrix components on ageing characteristics of transport between the choroid and the outer retina. AB - PURPOSE: To determine the relative influence of path length and matrix components on the movement of small solutes and water between the choroid and the outer retina. METHODS: Human and bovine Bruch's membrane-choroid (BC) tissue samples were mounted in modified Ussing chambers, and the diffusion of taurine and hydraulic conductivity (Lp) was determined. In humans, diffusion of taurine was determined as a function of age of the donor. The relative contribution of Bruch's membrane in the BC complex to transport processes was measured after its removal by laser ablation. Similarly, the effect of choroidal path length was determined. In humans, tracking the trend of age-related thinning provided samples of various path lengths. In young bovine animals (< or =2 years old), choroidal thickness was adjusted by laser ablation. RESULTS: Diffusion of taurine across human BC decreased linearly from 162.7 to 105.9 nanomoles/h per 3 mm between 10 and 90 years of age (P < 0.05). Ablation of Bruch's membrane increased diffusion of taurine from 129 to 287.9 nanomoles/h per 4 mm in human (donor age 55, 74, and 82 years; P < 0.005) but caused no statistically significant change in bovine BC. Diffusion of taurine across bovine BC was greater in samples with partially ablated choroid (218 nanomoles/h per 4 mm) than in normal control samples (128.75 nanomoles/h per 4 mm). Lp was not measurable in bovine samples after complete ablation of Bruch's membrane, but did not change significantly as the choroid thinned. CONCLUSIONS: The data suggest that both path length and matrix components contribute to the decline of diffusion of small solutes across BC with age. The importance of matrix components was also demonstrated in restricting the movement of water while choroidal thickness played little if any role. PMID- 15111608 TI - Effect of somatostatin on nitric oxide production in human retinal pigment epithelium cell cultures. AB - PURPOSE: To investigate the presence of somatostatin and its receptors (sst(1-5) receptors) and their possible involvement in the regulation of nitric oxide (NO) production in human RPE cell cultures. METHODS: Human RPE cells (D407) were used for all studies performed. Somatostatin levels were detected by radioimmunoassay, and RT-PCR and immunocytochemistry studies were performed to identify the somatostatin receptors (sst1-sst5). Radioligand binding assays were also performed examining the ability of certain somatostatin ligands (sst1, sst2, sst5) to compete for [125I]Tyr11 somatostatin binding. The presence of NO synthase in the cultures was assayed with NADPH-diaphorase cytochemistry, and RT PCR, and NO levels were assessed by examining the production of its stable metabolites NO2- and NO3- (NOx-). RESULTS: SRIF was detected in a concentration of 0.56 +/- 0.13 picomoles/mg protein. sst1, sst2, and sst5 mRNAs were detected, yet only sst2B and sst5 immunoreactivity was observed in human RPE cell cultures. sst1- and sst5- but not sst2-selective ligands displaced the specific [125I]Tyr11 somatostatin binding to RPE cell membranes. NADPH-diaphorase stain and iNOS mRNA were detected. SRIF and the sst2-selective analogue MK678 increased the levels of NOx- in a concentration-dependent manner. This increase was blocked by the sst2 antagonist CYN-154806 (Ac-4NO2-Phe-c(dCys-Tyr-dTrp-Lys-Thr-Cys)-dTyr-NH2). CONCLUSIONS: These results demonstrate the presence of somatostatin, and its receptors sst1, sst2B, and sst5 in human RPE cells and suggest an autocrine or paracrine role for somatostatin. Somatostatin's ability to regulate NO production, by activating sst2 receptors, provides a functional role of somatostatin in the RPE. PMID- 15111609 TI - Adrenomedullin affects two signal transduction pathways and the migration in retinal pigment epithelial cells. AB - PURPOSE: Adrenomedullin is a multifunctional regulatory peptide known to inhibit the migration of smooth muscle cells. In vitro studies were performed to identify whether adrenomedullin (ADM) also inhibits the migration of RPE cells. The aberrant behavior of these cells is an early event in proliferative vitreoretinopathy, and these studies were designed to determine how ADM acts on RPE cells at the second-messenger level. METHODS: Migration of cultured human RPE cells was determined by the Boyden chamber method, using 10% fetal calf serum (FCS) as a chemotactic factor. The attachment assay was performed on fibronectin, laminin, or poly-D-lysine-coated 96-well plates. RPE cells were incubated in PBS buffer with or without ADM for 15 minutes. Intracellular cAMP and cGMP changes were then measured by enzyme immunoassay (EIA). To determine the cytoplasmic free Ca2+ concentration ([Ca2+]i) response to ADM, fluo-3 AM-loaded RPE cells were imaged with a laser scanning confocal microscope, after stimulation with ADM (10( 12)-10(-7) M). RESULTS: ADM exhibited a concentration-dependent inhibition of FCS stimulated RPE cell migration. The maximum inhibitory effect of ADM, observed at 10(-7) M, on basal and FCS-induced RPE cell migration was approximately 53.8% and 43.8% of the control, respectively. Exogenously added ADM (10(-9)-10(-7) M) had no significant effect on RPE cell attachment on all tested substrates. ADM increased intracellular cAMP and decreased intracellular cGMP levels dose dependently (10(-10)-10(-7) M) in RPE. The maximum effect was observed at 10(-7) M. ADM also induced a [Ca2+]i decrease in a dose-dependent manner (10(-12)-10(-7) M). The maximum effect was observed at 0.1 micro M, at which point the level declined to 42.9% of the control. CONCLUSIONS: ADM inhibits the migration of RPE cells in vitro by a mechanism that involves the reciprocal upregulation of cAMP and downregulation of cGMP, in association with reductions in [Ca2+]i. ADM mediated fluctuations in [Ca2+]i, which are well known to be involved in cell migration, appear to be regulated in part by mechanisms involving cAMP synthase. Thus, it appears that ADM acts as a constitutive regulatory system to control aberrant RPE cell behavior and specific migration in response to inflammatory mediators. PMID- 15111610 TI - Effect of erythropoietin axotomy-induced apoptosis in rat retinal ganglion cells. AB - PURPOSE: Erythropoietin (EPO) modulates erythropoiesis by inhibiting apoptosis in erythrocyte progenitors. Recently, EPO has been shown to be protective in experimental models of mechanical trauma, neuroinflammation, cerebral and retinal ischemia, and even in a human stroke trial. However, little is known about EPO signal transduction in vivo and the usefulness of EPO in the prevention of the chronic, purely apoptotic neuronal cell death that contributes to vision loss in glaucoma and the progression of neurodegenerative diseases. METHODS: EPO's effects and signaling in the retinal ganglion cell axotomy paradigm were studied by Western blot analysis and immunohistochemistry, receptor expression was characterized in the retina before and after lesion. EPO was injected into the vitreous body to investigate neuroprotection of axotomized rat RGCs. Moreover, EPO's effects were studied in cultures of immunopurified retinal ganglion cells. Signal-transduction pathways transmitting neuroprotective EPO effects in vivo were characterized by the use of specific kinase inhibitors, immunohistochemistry, and Western blot analysis. RESULTS: EPO receptors (EPORs) were expressed on RGC somata and dendrites in vivo. EPOR expression did not significantly change after axotomy. Application of EPO prevented death of neurotrophic-factor-deprived immunopurified rat RGCs in vitro, rescued axotomized RGCs in vivo, and prevented caspase-3 activation. EPO-induced Akt phosphorylation and survival-promoting EPO effects were completely abolished by inhibition of PI 3-kinase. EPO neuroprotection followed a bell-shaped dose-response curve in vitro and in vivo, whereas toxic EPO effects were never observed, even at high concentrations. CONCLUSIONS: These data support a potential role for EPO as a therapeutic molecule against predominantly apoptotic neuronal cell death in the context of glaucoma or neurodegenerative diseases and delineate the PI-3-K/Akt pathway as the predominant mediator of EPO neuroprotection in this in vivo paradigm of neuronal cell death. PMID- 15111611 TI - Protection by pyruvate of rat retinal cells against zinc toxicity in vitro, and pressure-induced ischemia in vivo. AB - PURPOSE: To examine whether zinc accumulation occurs during retinal neuronal death after pressure-induced ischemia in rats and whether pyruvate protects against such death. METHODS: To induce transient retinal ischemia, intraocular pressure was increased above systolic pressure for 65 minutes. Pyruvate was administered through the tail vein for 12 hours after ischemia to determine its effect on degeneration of retinal neurons. Retinas were removed and sectioned, and zinc accumulation was visualized with N-(6-methoxy-8-quinolyul)-p carboxybenzoyl-sylphonamide (TFL-Zn) fluorescence microscopy, and neuronal death was determined with acid fuchsin staining. For in vitro studies, retinal cell cultures were prepared from newborn rat pups and used for experiments at days in vitro (DIV) 7 to 10. RESULTS: After retinal ischemia, staining revealed that most zinc-accumulating neurons were injured neurons, suggesting that endogenous zinc may contribute to ischemic neuronal death in the retina. In vitro studies showed that 15 minutes of exposure to 300 to 500 microM zinc resulted in the death of a substantial number of retinal cells in culture, and that this death was preceded by poly(ADP-ribose) polymerase (PARP)-mediated depletion of nicotinamide-adenine dinucleotide (NAD+) and adenosine triphosphate (ATP). Pyruvate, but not lactate, protected against this zinc-induced cell death in vitro. Consistent with this finding, in vivo studies showed that compared with control rats, pyruvate-treated rats had a substantial reduction in the number of cells showing signs of cell death. CONCLUSIONS: The present results suggest endogenous zinc contributes to retinal cell death after ischemia. Pyruvate potently protected against zinc toxicity in cultured rat retinal cells and reduced ischemia-induced cell death in rat retinas. PMID- 15111612 TI - Acetylcholine protection of adult pig retinal ganglion cells from glutamate induced excitotoxicity. AB - PURPOSE: To determine which glutamate receptor (GluR) subtypes are responsible for glutamate-induced excitotoxicity in cultured adult pig retinal ganglion cells (RGCs) and to characterize the neuroprotective effect of acetylcholine (ACh) on pig RGCs. METHODS: Adult pig RGCs were isolated from other retinal tissue by a modified panning technique using Thy 1.1 antibody. Isolated RGCs were cultured in control media and media containing: glutamate, NMDA, or KA; glutamate and CNQX, MK-801, or AP-7; ACh, nicotine or muscarine; ACh and alpha-bungarotoxin (Bgt) or methyllycaconitine (MLA); and glutamate and choline or glutamate, choline, and MLA. To determine cell viability, cells were loaded with calcein and counted. RESULTS: Ninety-eight percent of isolated cells were immunolabeled with Thy 1.1 antibody. Chronic exposure to 500 microM glutamate decreased the number of surviving large and small RGCs, compared to control conditions. This glutamate induced excitotoxicity was mediated through both NMDA and non-NMDA GluRs. In neuroprotective studies, ACh, nicotine, and choline significantly reduced glutamate-induced excitotoxicity in adult pig RGCs through alpha-Bgt-sensitive nicotinic ACh receptors (nAChRs). DISCUSSION: This was the first report of a modified panning technique to isolate adult pig RGCs. Cell viability was relatively high using this method, and both large and small RGCs grew extensive neurites in culture. The finding that both NMDA and non-NMDA GluRs were involved in glutamate-induced excitotoxicity suggests that isolated pig RGCs provide a good model for glaucoma. In addition, activation of AChRs may be useful in protecting RGC from excitotoxic insults occurring in neurodegenerative diseases such as glaucoma. PMID- 15111613 TI - Localization of collagen XVIII and the endostatin portion of collagen XVIII in aged human control eyes and eyes with age-related macular degeneration. AB - PURPOSE: Endostatin, a C-terminal fragment of collagen XVIII (coll XVIII) formed by proteolysis, specifically inhibits endothelial cell migration and proliferation in vitro and potently inhibits angiogenesis and tumor growth in vivo. The purpose of this study was to examine the immunolocalization of endostatin and coll XVIII in the retina and choroid of human donor tissue sections from aged control donor eyes and to determine whether the localization or relative levels are changed in age-related macular degeneration (AMD). METHODS: Ocular tissues were obtained from six aged control donors (age range, 75 86 years; mean age, 80.5 years) without evidence or history of chorioretinal disease and from nine donors with AMD (age range, 74-105 years; mean age, 88.6 years). Tissues were cryopreserved, and streptavidin alkaline phosphatase immunohistochemistry was performed with goat anti-human and mouse anti-human endostatin antibodies and rabbit anti-mouse coll XVIII. Blood vessels were identified with mouse anti-human CD-34 antibody in adjacent sections. Pigment in RPE and choroidal melanocytes was bleached. Three independent observers scored the immunohistochemical reaction product. RESULTS: In aged control eyes, coll XVIII and endostatin (the endostatin portion of coll XVIII) immunoreactivity was observed in large retinal blood vessels and in capillaries in some individuals, but the internal limiting membrane (ILM) had the most intense retinal immunostaining. There was no significant difference in immunoreactivity to both antibodies in retinal blood vessels in aged control eyes. In the choroid, endostatin and coll XVIII were localized to blood vessels, Bruch's membrane, and RPE basal lamina. AMD retina and choroid had a similar pattern and intensity of coll XVIII immunostaining, as observed in control eyes but reaction product was more diffuse in the choroid. Endostatin immunoreactivity was significantly higher in ILM (P = 0.037) in AMD retina and significantly lower in the choriocapillaris, Bruch's membrane, and RPE basal lamina of AMD choroids (P < 0.05) and completely negative in some areas of AMD choroids. CONCLUSIONS: These data suggest that reduced levels of the endostatin portion of coll XVIII in Bruch's membrane, RPE basal lamina, intercapillary septa, and choriocapillaris in eyes with AMD may be permissive for choroidal neovascularization. PMID- 15111614 TI - Nuclear translocation of glyceraldehyde-3-phosphate dehydrogenase: a role in high glucose-induced apoptosis in retinal Muller cells. AB - PURPOSE: A recent study demonstrated that retinal Muller cells undergo hyperglycemia-induced apoptosis in vitro. Translocation of glyceraldehyde-3 phosphate dehydrogenase (GAPDH) from the cytosol to the nucleus is a critical step in the induction of apoptosis in neuronal cells. R-(-)-deprenyl prevents nuclear translocation of GAPDH and subsequent apoptosis in neuronal cells. In this study, the role of nuclear translocation of GAPDH in hyperglycemia-induced apoptosis in retinal Muller cells and the ability of R-(-)-deprenyl to inhibit the translocation of GAPDH and apoptosis were investigated. METHODS: Transformed rat Muller cells (rMC-1) and isolated human Muller cells were cultured in normal glucose, high glucose, and high glucose plus R-(-)-deprenyl for up to 5 days. Subcellular distribution of GAPDH was determined in vitro and in vivo by immunocytochemistry. Apoptosis in tissue cultures was determined by annexin-V staining and caspase-3 activity. RESULTS: Hyperglycemia significantly increased the amount of GAPDH protein in the nucleus above normal within the first 48 hours in rMC-1 and human Muller cells. The addition of R-(-)-deprenyl to these cells incubated in high glucose reduced the amount of GAPDH protein in the nucleus and decreased hyperglycemia-induced apoptosis in both cell types. In vivo studies confirmed the accumulation of GAPDH in nuclei of Muller cells in diabetes. CONCLUSIONS: The nuclear translocation of GAPDH in rMC-1 and human Muller cells is closely associated with the induction of apoptosis. R-(-)-deprenyl inhibits nuclear accumulation of GAPDH and subsequent apoptosis in these cells. Therefore, R-(-)-deprenyl offers a strategy to explore the role of GAPDH translocation into the nucleus in the development of diabetic retinopathy. PMID- 15111615 TI - Topographic and age-dependent expression of heme oxygenase-1 and catalase in the human retinal pigment epithelium. AB - PURPOSE: To investigate the hypothesis that there are topographic and age-related changes in the expression of heme oxygenase (HO)-1 and catalase in the RPE. METHODS: Cryosections of the macula and periphery of human eyes (n = 18; aged 27 87 years) were subjected to a high-sensitivity digoxigenin (DIG)-labeled cRNA in situ hybridization protocol to determine the expression of HO-1 and catalase. The immunoreactivity of HO-1 and catalase were also investigated in the same sample set. Specimens were examined by light microscopy, and images were captured with a digital camera. The total number of RPE cells and HO-1- and catalase-labeled RPE cells was counted in each section, and the ratio of labeled RPE cells to total RPE cells was calculated in both the macular and the peripheral regions of each donor eye. RESULTS: There was a mosaic pattern of mRNA and protein expression of HO-1 and catalase in macular and peripheral RPE. Topographical differences in the expression of HO-1 at the mRNA level and catalase at both the mRNA and protein levels was also observed. The topographical differences between the expression of HO-1 in the macula and periphery protein were not statistically significant but showed similar trends. For HO-1, the only significant age-related decline in expression was observed in the macula and periphery. Expression of HO-1 at the protein level and that of catalase at both the mRNA and protein levels showed no significant decline with age. CONCLUSIONS: There is a possible age-related decline in HO-1 expression, whereas catalase expression remains unchanged with aging. Both exhibit mosaic patterns in the RPE monolayer. PMID- 15111616 TI - Spatial vision deficits in infants and children with Down syndrome. AB - PURPOSE: Infants and children with Down syndrome show reduced visual acuity and contrast sensitivity when tested with conventional behavioral techniques. These results may reflect sensory deficits of optical or neural origin or a loss of performance in mechanisms responsible for generating the behavioral response. The purpose of this study was to compare objective acuity and contrast sensitivity measurements recorded with visual-evoked potentials (VEPs), with behavioral clinical test results in a group of children with Down syndrome and a group of control subjects. The goal was to determine whether children with Down syndrome still have a sensory deficit when tested using a procedure that is less cognitively demanding than conventional tests. METHODS: The subject group comprised 58 children with Down syndrome and 44 control subjects, aged 3 months to 14.15 years. Visual acuity and contrast sensitivity were measured with steady state, swept VEPs and behavioral techniques. VEP acuity was obtained from 36 children with Down syndrome and 40 control subjects, and behavioral acuity from 54 children with Down syndrome and 35 control subjects. VEP contrast sensitivity was measured in 24 children with Down syndrome and 34 control subjects, and behavioral contrast sensitivity in 42 children with Down syndrome and 25 control subjects. Group differences in visual acuity and contrast sensitivity were analyzed with an analysis of covariance (ANCOVA), with age as a covariate. RESULTS: Visual acuity thresholds were significantly lower in the group with Down syndrome than in the control group. This was true for both VEP (P < 0.01) and behavioral measures (P < 0.01). The Down syndrome group also had reduced contrast sensitivity when compared with the control subjects, for VEP contrast sensitivity (P < 0.01) and behavioral contrast sensitivity (P < 0.01). The group differences remained when children with ophthalmic anomalies were excluded from the analysis. CONCLUSIONS: The reduced visual acuity and contrast sensitivity in the Down syndrome group support the idea of an underlying sensory deficit in the visual system in Down syndrome. PMID- 15111617 TI - Magnitude of response with myeloma frontline therapy does not predict outcome: importance of time to progression in southwest oncology group chemotherapy trials. AB - PURPOSE: Four Southwest Oncology Group (SWOG) standard-dose chemotherapy protocols for multiple myeloma (MM) initiated between 1982 and 1992 were evaluated. The purpose was to clarify the predictive value of specific levels of myeloma-associated monoclonal protein reduction and time to first progression using mature data sets. PATIENTS AND METHODS: Study data on 1,555 eligible previously untreated patients with MM enrolled onto SWOG phase III trials 8229, 8624, 9028, and 9210 were used in these analyses. Six-month and 12-month landmark analyses were performed to evaluate the outcome for patients in each response category. RESULTS: The overall and event-free survivals for the four protocols combined were 33 months and 18 months, respectively. Using 6- and 12-month landmarks, the median survivals of 30 to 35 months were not different for responders (> or = 50% and > or = 75% regression) versus nonresponders in patients without disease progression before the landmarks. Conversely, at the 6- and 12-month landmarks, the median survivals for patients who had experienced disease progression were 13 and 15 months, respectively, versus a 34-month median for patients who did not experience progression. Using the Cox survival model, with response and progression considered as time-dependent covariates, survival duration was influenced more by the occurrence of progression than by the occurrence of response. CONCLUSION: The magnitude of response, as a single variable, does not predict survival duration. Patients with response and stable disease have equivalent outcome. Only patients with progressive disease have a poorer outcome. The best indicator of survival is time to first progression. PMID- 15111618 TI - High-dose chemotherapy with autologous hematopoietic stem-cell support compared with standard-dose chemotherapy in breast cancer patients with 10 or more positive lymph nodes: first results of a randomized trial. AB - PURPOSE: Investigation of high-dose chemotherapy (HD-CT) followed by autologous hematopoietic stem-cell support compared with standard-dose chemotherapy (SD-CT) as adjuvant treatment in patients with primary breast cancer and 10 or more positive axillary lymph nodes. PATIENTS AND METHODS: Between November 1993 and September 2000, 307 patients were randomized to receive (following four cycles of epirubicin 90 mg/m(2) and cyclophosphamide 600 mg/m(2), intravenously every 21 days) either HD-CT of cyclophosphamide 1500 mg/m(2), thiotepa 150 mg/m(2), and mitoxantrone 10 mg/m(2), intravenously for 4 consecutive days followed by stem cell support; or SD-CT in three cycles of cyclophosphamide 500 mg/m(2), methotrexate 40 mg/m(2), and fluorouracil 600 mg/m(2) intravenously on days 1 and 8, every 28 days. The primary end point was event-free survival. RESULTS: After a median follow-up of 3.8 years, 144 events with respect to event-free survival have been observed (HD-CT: 63 events; SD-CT: 81 events). The first event of failure (HD-CT v SD-CT) was an isolated locoregional recurrence (nine v 11), a distant failure (52 v 68), and death without recurrence (two v two). The estimated relative risk of HD-CT versus SD-CT was 0.75 (95% CI, 0.54 to 1.06; P =.095). Overall survival showed no difference (HD-CT: 40 deaths; SD-CT: 49 deaths). CONCLUSION: There was a trend in favor of HD-CT with respect to event free survival, but without statistical significance. Further follow-up and a meta analysis of all randomized studies will reveal the effect of HD-CT as compared with SD-CT as adjuvant treatment in high-risk primary breast cancer. PMID- 15111619 TI - Are chemotherapy response rates related to treatment-induced survival prolongations in patients with advanced cancer? AB - PURPOSE: Patients with incurable cancer are faced with difficult decisions regarding whether to take chemotherapy in an attempt to preserve the quality and/or prolong the quantity of their lives. The average prolongation in survival with chemotherapy compared with best supportive care has not been well described. METHODS: We performed a literature search using PUBMED combined with expert inquiry to identify trials comparing cytotoxic chemotherapy with best supportive care. Twenty-five randomized, controlled clinical trials comparing cytotoxic chemotherapy with best supportive care were identified. Sixteen trials (64%) were in patients with non-small-cell lung cancer (NSCLC). Data were extracted and analyzed. RESULTS: Sufficient data for statistical modeling were available for NSCLC trials. The mean sample size of the NSCLC trials was 175 patients. Response rates in the treatment arms for NSCLC ranged from 7% to 42%. A relationship between response rate and survival was observed for NSCLC. The estimated relationship for NSCLC suggested that each 3.3% increase in response rate correlated, on average, with a 1-week increase in median survival, and each 2% increase in response rate correlated, on average, with a 1% increase in 1-year survival. The mean increase in 1-year survival for trials of agents with at least a 20% response rate in NSCLC was 16%. Formulas are provided to help estimate how a given response rate may effect median and 1-year survival relative to best supportive care alone for NSCLC. CONCLUSION: We found a relationship between response rate and both median and 1-year survival in NSCLC. This information may help oncologists estimate how an NSCLC chemotherapy regimen with a given response rate can, on average, impact survival relative to supportive care alone. PMID- 15111620 TI - What is the role of high-dose chemotherapy in the era of targeted therapies? PMID- 15111621 TI - Dioxin-induced immortalization of normal human keratinocytes and silencing of p53 and p16INK4a. AB - Dioxin, a potent tumor promoter, activates the aryl hydrocarbon receptor (AHR), a ligand-activated transcription factor belonging to the basic helix-loop-helix-PAS family, to enhance tumorigenesis via unknown mechanisms. We undertook this study to determine the mechanisms underlying the impact of dioxin on cell fate, in particular senescence that occurs in normal human cells and is considered to play important tumor suppressive function. We have previously shown that in primary human keratinocytes, dioxin attenuates senescence while retaining the proliferative capacity and represses expression of the tumor suppressors, p16(INK4a) and p53. Here, we show that repression of p16(INK4a) and p53 transcriptional activity by dioxin absolutely requires the AHR and is accompanied by promoter methylation. Furthermore, dioxin alone is sufficient to immortalize normal human keratinocytes. Our data introduce a previously unrecognized regulatory pathway, that of the AHR, that impacts senescence. More importantly, this is the first report of a tumor promoter capable of inhibiting senescence in a receptor mediated manner and introduces a novel mechanism by which this carcinogen may contribute to human malignancies. PMID- 15111622 TI - Cbf1p is required for chromatin remodeling at promoter-proximal CACGTG motifs in yeast. AB - Cbf1p is a basic-helix-loop-helix-zipper protein of Saccharomyces cerevisiae required for the function of centromeres and MET gene promoters, where it binds DNA via the consensus core motif CACRTG (R = A or G). At MET genes Cbf1p appears to function in both activator recruitment and chromatin-remodeling. Cbf1p has been implicated in the regulation of other genes, and CACRTG motifs are common in potential gene regulatory DNA. A recent genome-wide location analysis showed that the majority of intergenic CACGTG palindromes are bound by Cbf1p. Here we tested whether all potential Cbf1p binding motifs in the yeast genome are likely to be bound by Cbf1p using chromatin immunoprecipitation. We also tested which of the motifs are actually functional by assaying for Cbf1p-dependent chromatin remodeling. We show that Cbf1p binding and activity is restricted to palindromic CACGTG motifs in promoter-proximal regions. Cbf1p does not function through CACGTG motifs that occur in promoter-distal locations within coding regions nor where CACATG motifs occur alone except at centromeres. Cbf1p can be made to function at promoter-distal CACGTG motifs by overexpression, suggesting that the concentration of Cbf1p is normally limiting for binding and is biased to gene regulatory DNA by interactions with other factors. We conclude that Cbf1p is required for normal nucleosome positioning wherever the CACGTG motif occurs in gene regulatory DNA. Cbf1p has been shown to interact with the chromatin remodeling ATPase Isw1p. Here we show that recruitment of Isw1p by Cbf1p is likely to be general but that Isw1p is only partially required for Cbf1p dependent chromatin structures. PMID- 15111624 TI - Challenges and opportunities. PMID- 15111625 TI - Henry Klein--a forgotten icon? PMID- 15111623 TI - Structural model of MD-2 and functional role of its basic amino acid clusters involved in cellular lipopolysaccharide recognition. AB - The receptor complex resulting from association of MD-2 and the ectodomain of Toll-like receptor 4 (TLR4) mediates lipopolysaccharide (LPS) signal transduction across the cell membrane. We prepared a tertiary structure model of MD-2, based on the known structures of homologous lipid-binding proteins. Analysis of circular dichroic spectra of purified bacterially expressed MD-2 indicates high content of beta-type secondary structure, in agreement with the structural model. Bacterially expressed MD-2 was able to confer LPS responsiveness to cells expressing TLR4 despite lacking glycosylation. We identified several clusters of basic residues on the surface of MD-2. Mutation of each of two clusters encompassing the residues Lys(89)-Arg(90)-Lys(91) and Lys(125)-Lys(125) significantly decreased the signal transduction of the respective MD-2 mutants either upon co-expression with TLR4 or upon addition as soluble protein into the supernatant of cells overexpressing TLR4. These basic clusters lie at the edge of the beta-sheet sandwich, which in cholesterol-binding protein connected to Niemann-Pick disease C2 (NPC2), dust mite allergen Der p2, and ganglioside GM2 activator protein form a hydrophobic pocket. In contrast, mutation of another basic cluster composed of Arg(69)-Lys(72), which according to the model lies further apart from the hydrophobic pocket only weakly decreased MD-2 activity. Furthermore, addition of the peptide, comprising the surface loop between Cys(95) and Cys(105), predicted by model, particularly in oxidized form, decreased LPS induced production of tumor necrosis factor alpha and interleukin-8 upon application to monocytic cells and fibroblasts, respectively, supporting its involvement in LPS signaling. Our structural model of MD-2 is corroborated by biochemical analysis and contributes to the unraveling of molecular interactions in LPS recognition. PMID- 15111626 TI - Novel mutations in the cathepsin C gene in patients with pre-pubertal aggressive periodontitis and Papillon-Lefevre syndrome. AB - Aggressive periodontitis (AP) in pre-pubertal children is often associated with genetic disorders like Papillon-Lefevre syndrome (PLS). PLS is caused by mutations in the cathepsin C (CTSC) gene. We report a novel CTSC mutation (c.566 572del) in an otherwise healthy AP child and two novel compound heterozygous mutations (c.947T>G, c.1268G>C) in a PLS patient. We conclude that at least a subset of pre-pubertal AP is due to CTSC mutations and therefore may be an allelic variant of PLS. PMID- 15111627 TI - Increase in HIV receptors/co-receptors/alpha-defensins in inflamed human gingiva. AB - Transmission of HIV-1 through the oral cavity is considered to be a rare event. To identify factors in resistance/susceptibility to oral HIV-1 infection, we analyzed expression in human gingiva of HIV-1 receptors Langerin, DC-SIGN, MR, and GalCer, HIV-1 co-receptors CCCR5, CXCR4, and anti-microbial protein alpha defensin-1. Our results show that healthy gingiva is infiltrated with cells expressing all HIV-1 receptors tested; however, there are very few CCR5(+) cells and a complete absence of CXCR4(+) cells in the lamina propria. In chronic periodontitis (CP), DC-SIGN, MR, CXCR4, and CCR5 increase, but this was accompanied by a ten-fold increase in alpha-defensin-1 mRNA. The CCR5(+) cells were revealed to be T-cells, macrophages, and dermal dendritic cells. Moreover, epithelial expression of GalCer and CXCR4 together was not apical and showed no trend with underlying inflammation. Thus, low expression of HIV-1 co-receptors in health and high expression of alpha-defensin during CP may comprise endogenous factors that provide protection from oral HIV-1 infection. PMID- 15111628 TI - Amelogenin p.M1T and p.W4S mutations underlying hypoplastic X-linked amelogenesis imperfecta. AB - Mutations in the human amelogenin gene (AMELX, Xp22.3) cause a phenotypically diverse set of inherited enamel malformations. We hypothesize that the effects of specific mutations on amelogenin protein structure and expression will correlate with the enamel phenotype, clarify amelogenin structure/function relationships, and improve the clinical diagnosis of X-linked amelogenesis imperfecta (AI). We have identified two kindreds with X-linked AI and characterized the AMELX mutations underlying their AI phenotypes. The two missense mutations are both in exon 2 and affect the translation initiation codon and/or the secretion of amelogenin (p.M1T and p.W4S), resulting in hypoplastic enamel. Primary anterior teeth from affected females with the p.M1T mutation were characterized by light and scanning electron microscopy. The thin enamel had defective prism organization, and the surface was rough and pitted. Dentin was normal. The severity of the enamel phenotype correlated with the predicted effects of the mutations on amelogenin expression and secretion. PMID- 15111629 TI - Mast cells in human periodontal disease. AB - Recently, mast cells have been shown to produce cytokines which can direct the development of T-cell subsets. The aim of the present study was to determine the relationship between mast cells and the Th1/Th2 response in human periodontal disease. Tryptase+ mast cell numbers were decreased in chronic periodontitis tissues compared with healthy/gingivitis lesions. Lower numbers of c-kit+ cells, which remained constant regardless of clinical status, indicate that there may be no increased migration of mast cells into periodontal disease lesions. While there were no differences in IgG2+ or IgG4+ cell numbers in healthy/gingivitis samples, there was an increase in IgG4+ cells compared with IgG2+ cells in periodontitis lesions, numbers increasing with disease severity. This suggests a predominance of Th2 cells in periodontitis, although mast cells may not be the source of Th2-inducing cytokines. PMID- 15111630 TI - Relationship between oral sensitivity and masticatory performance. AB - The size of a bolus determines how it will be manipulated in the mouth and swallowed. We hypothesized that mucosal sensitivity would be important for masticatory function. The accuracy of solid object size perception, spatial acuity, and food particle size reduction during mastication were measured in 22 healthy adults with/without topical anesthesia of their oral mucosa. Topical anesthesia had no effect on the perception of sphere sizes, but significantly reduced spatial sensitivity. Without anesthesia, there was a correlation between an individual's ability to perceive the sizes of steel spheres (diameter, 4-9 mm) and the sizes of food particles chewed for 15 cycles and at swallowing. There was no correlation between spatial sensitivity and food particle size. We suggest that the stimuli used to test two-point discrimination stimulates only superficial receptors, which involve light touch and are easily anesthetized, while the spheres might excite more deeply-set receptors. The latter appear to be more important for masticatory performance and swallowing. PMID- 15111631 TI - Oral pilocarpine for treatment of opioid-induced oral dryness in healthy adults. AB - Pilocarpine induces a profuse flow of saliva when administered orally, but effects on drug-induced oral dryness have not been examined. The aim of this trial was to investigate if pilocarpine increases production of saliva in individuals suffering from dry mouth due to treatment with opioids. Sixty-five individuals were enrolled in a randomized, double-blind, placebo-controlled trial. The subjects received tramadol (50 mg t.d.s.) to induce oral dryness, and were thereafter assigned to one of three groups. Secretion rate of saliva was measured before and after tramadol, and after the oral administration of pilocarpine (5 mg), placebo, or no treatment. Baseline characteristics did not differ among the groups (mean +/- SEM: 0.37 +/- 0.06 mL/min), and tramadol lowered the secretion at the same level in all groups (0.15 +/- 0.02 mL/min). Pilocarpine increased the flow above that observed with placebo (0.66 +/- 0.19 vs. 0.15 +/- 0.02 mL/min). Thus, pilocarpine re-establishes the flow of saliva in the state of tramadol-induced oral dryness. PMID- 15111632 TI - Quantitative polygraphic controlled study on efficacy and safety of oral splint devices in tooth-grinding subjects. AB - The efficacy of occlusal splints in diminishing muscle activity and tooth grinding damage remains controversial. The objective of this study was to compare the efficacy and safety of an occlusal splint (OS) vs. a palatal control device (PCD). Nine subjects with sleep bruxism (SB) participated in this randomized study. Sleep laboratory recordings were made on the second night to establish baseline data. Patients then wore each of the splints in the sleep laboratory for recording nights three and four, two weeks apart, according to a crossover design. A statistically significant reduction in the number of SB episodes per hour (decrease of 41%, p = 0.05) and SB bursts per hour (decrease of 40%, p < 0.05) was observed with the two devices. Both oral devices also showed 50% fewer episodes with grinding noise (p = 0.06). No difference was observed between the devices. Moreover, no changes in respiratory variables were observed. Both devices reduced muscle activity associated with SB. PMID- 15111633 TI - The frictional coefficient of the temporomandibular joint and its dependency on the magnitude and duration of joint loading. AB - In synovial joints, friction between articular surfaces leads to shear stress within the cartilaginous tissue, which might result in tissue rupture and failure. Joint friction depends on synovial lubrication of the articular surfaces, which can be altered due to compressive loading. Therefore, we hypothesized that the frictional coefficient of the temporomandibular joint (TMJ) is affected by the magnitude and duration of loading. We tested this by measuring the frictional coefficient in 20 intact porcine TMJs using a pendulum-type friction tester. The mean frictional coefficient was 0.0145 (SD 0.0027) after a constant loading of 50 N during 5 sec. The frictional coefficient increased with the length of the preceding loading duration and exceeded 0.0220 (SD 0.0014) after 1 hr. Application of larger loading (80 N) resulted in significantly larger frictional coefficients. In conclusion, the frictional coefficient in the TMJ was proportional to the magnitude and duration of joint loading. PMID- 15111634 TI - Root-end filling materials alter fibroblast differentiation. AB - Root-end filling materials are commonly used following endodontic surgical procedures; however, their effect on adjacent soft tissues is poorly understood. We predict that, due to the differences in their chemical composition, these materials will have profoundly different effects on the survival and differentiation of fibroblasts. Many of the root-end filling materials examined were initially cytotoxic to both PDL and gingival fibroblasts in co-culture experiments; however, this was reduced after the materials were washed in either mineral trioxide aggregate (MTA) or hybrid ionomere composite resin (HICR) for 2 wks. Additionally, PDL fibroblasts displayed enhanced proliferation on MTA and survival on amalgam when compared with gingival fibroblasts. MTA preferentially induced alkaline phosphatase expression and activity in both PDL and gingival fibroblasts. In contrast, HICR inhibited alkaline phosphatase expression and activity. In addition, MTA and HICR repressed pleiotrophin in PDL fibroblasts, while HICR repressed periostin in both fibroblasts. Thus, root-end filling materials differentially affect periodontal fibroblast differentiation. ABBREVIATIONS: mineral trioxide aggregate (MTA), zinc-oxide eugenol cement (ZOEC), hybrid ionomer composite resin (HICR), reverse-transcriptase polymerase chain-reaction (RT-PCR). PMID- 15111636 TI - The influence of specimen attachment and dimension on microtensile strength. AB - The higher microtensile bond strength values found for specimens with a smaller cross-sectional area are often explained by the lower occurrence of internal defects and surface flaws. We hypothesized that this aberrant behavior is mainly caused by the lateral way of attachment of the specimens to the testing device, which makes the strength dependent on the thickness. This study showed that composite bars of 1x1x10, 1x2x10, and 1x3x10mm attached at their 1-mm-wide side (situation A) fractured at loads of the same magnitude, as a result of which the microtensile strength ( micro TS), calculated as F/A (force at fracture/cross sectional area), significantly increased for specimens with decreasing thickness. Attachment at the 1-, 2-, or 3-mm-wide side (situation B) resulted in equal micro TS values (P > 0.05). Finite element analysis showed different stress patterns for situation A, but comparable patterns for situation B. Both situations showed the same maximum stress at fracture. PMID- 15111635 TI - Collagen degradation in endodontically treated teeth after clinical function. AB - Endodontically treated teeth restored with posts are susceptible to coronal leakage after long-term function. We hypothesize that demineralized collagen matrices (DCMs) created in dentin by acidic zinc phosphate cement within the dowel spaces degrade with time. Forty-two post-restored teeth were extracted after three periods of clinical service and were examined, by means of scanning and transmission electron microscopy, for the status of the DCMs. SEM revealed a progressive degradation of the DCMs, becoming less dense after 3 to 5 years, losing structural integrity after 6 to 9 years, and partially disappearing after 10 to 12 years. TEM revealed evidence of collagenolytic activity within the DCMs, with loss of cross-banding and unraveling into microfibrils, and gelatinolytic activity that resulted in disintegration of the microfibrils. Bacterial colonization and the release of bacterial enzymes and of host-derived matrix metalloproteinases may contribute to the degradation of collagen fibrils in root dentin after clinical function. PMID- 15111637 TI - Nanocrystalline tetracalcium phosphate cement. AB - Calcium hydroxide cements can lack long-term stability and achieve sustained release by matrix-controlled diffusion of hydroxyl ions. Tetracalcium phosphate (TTCP) hydrolyzes slowly to form calcium hydroxide and a thin insoluble apatite layer that prevents further reaction. In this study, mechanical amorphization was used to create a setting calcium-hydroxide-releasing cement from TTCP. The effect of high-energy ball milling of TTCP on the mechanical properties of the cement was investigated. X-ray diffraction data were used to determine the phase composition of the set cements. An accelerated in vitro test compared pH of water after prolonged boiling of nanocrystalline TTCP cements and a calcium salicylate material. As milling time increased, cement compressive strength and degree of conversion increased. Hydroxyl ion release from the cement was comparable with that from a calcium salicylate material. This new cement system offers the antimicrobial potential of calcium salicylate materials combined with the long term stability of insoluble apatite cements. PMID- 15111638 TI - Induction of tolerance by Porphyromonas gingivalis on APCS: a mechanism implicated in periodontal infection. AB - The periodontal pathogen Porphyromonas gingivalis (Pg) is a potent inducer of the production of pro-inflammatory cytokines by neutrophils, monocytes, and macrophages, and can desensitize immune cells in vitro and in vivo. We analyzed the ability of Pg lipopolysaccharide (LPS) to induce endotoxin tolerance. Treatment of dendritic cells (DC), the human macrophage cell line THP-1, and monocytes (antigen-presenting cells, APC) with Pg.LPS inhibited APC maturation assessed by CD80 and CD86 expression, and inhibited chemokine (CCL3 and CCL5) production. Pre-treatment with glucocorticoids (GC) and interleukin-10 (IL-10) abolished the effect of Pg.LPS on CD80, CD83, and CD86, and on CCL3 and CCL5 production. We also showed that Pg.LPS enhanced the tolerogenic properties of APCs and up-regulated ILT-3 and B7-H1 expression. PMID- 15111639 TI - Indian hedgehog: a mechanotransduction mediator in condylar cartilage. AB - Indian hedgehog (Ihh) is a critical mediator transducing mechanical signals to stimulate chondrocyte proliferation. To clarify the cellular signal transduction pathway that senses and converts mechanical signals into tissue growth in mandibular condyle, we evaluated Ihh expression and its relation to the kinetics of replicating mesenchymal cells in condylar cartilage during natural growth and mandibular advancement. Thirty-five-day-old Sprague-Dawley rats were fitted with functional appliances. Experimental animals with matched controls were doubly labeled with iododeoxyuridine and bromodeoxyuridine so that we could evaluate the cycles of the proliferative mesenchymal cells. Mandibular advancement triggered Ihh expression in condylar cartilage. A higher level of Ihh expression coincided with the increase of the replicating mesenchymal cells' population and the shortening of the turnover time. These findings suggested that Ihh acts as a mediator of mechanotransduction that converts mechanical signals resulting from anterior mandibular displacement to stimulate cellular proliferation in condylar cartilage. PMID- 15111640 TI - Inorganic lead exposure in the rat activates striatal cFOS expression at lower blood levels and inhibits amphetamine-induced cFOS expression at higher blood levels. AB - The impact of inorganic lead exposure on dopamine (DA) neurotransmission in the basal ganglia was examined. Amphetamine (AMPH)-induced cFOS immunoreactivity (cFOS-IR) in the striatum was determined after a 3-week exposure to lead acetate (0, 50, or 250 ppm). On the 21st day of lead exposure, rats were challenged with AMPH (4 mg/kg i.p.) or saline vehicle (Veh) and were assayed for presence of cFOS IR. In the untreated control (Con) group, AMPH challenge (Con/AMPH) increased cFOS-IR expression by approximately 35-fold over Veh challenge (Con/Veh) (P < 0.01). In the Pb50/Veh group, cFOS-IR expression was approximately 7-fold greater than in the Con/Veh group (P < 0.05). Given that there was negligible cFOS-IR expression in the Con/Veh group, this indicates that the Pb50 exposure induced cFOS expression. The increase in cFOS-IR in the Pb50/AMPH was also significant (P < 0.01), but it was not different from the Con/AMPH (P > 0.20). Neither the Pb250/Veh group nor the Pb250/AMPH group had a significant increase in cFOS-IR relative to Con/Veh (P > 0.20). These results indicate that chronic 50 ppm lead exposure induced a low but statistically significantly level of cFOS gene activation and that it did not affect the AMPH-induced cFOS activation. However, chronic 250 ppm lead exposure inhibited AMPH-induced activation of cFOS in the striatum by about 89%. Therefore, lead is capable of both activating cFOS expression at low levels of exposure (mean blood lead level 21.6 +/- 1.9 microg/dl) and inhibiting AMPH-induced cFOS expression at higher levels of exposure (mean blood lead level 47.4 +/- 2.6 microg/dl). PMID- 15111641 TI - ITH4012 (ethyl 5-amino-6,7,8,9-tetrahydro-2-methyl-4 phenylbenzol[1,8]naphthyridine-3-carboxylate), a novel acetylcholinesterase inhibitor with "calcium promotor" and neuroprotective properties. AB - Ethyl 5-amino-6,7,8,9-tetrahydro-2-methyl-4-phenylbenzol[1,8] naphthyridine-3 carboxylate (ITH4012) is a novel tacrine derivative that can reduce cell death induced by various compounds with different mechanisms of action, such as thapsigargin (reticular stress), H2O2 (free radicals), and veratridine (calcium overload), in bovine chromaffin cell. Cell viability, quantified as lactic dehydrogenase release, was significantly reduced by ITH4012 at concentrations ranging from 0.01 to 3 microM. In the human neuroblastoma cell line SH-SY5Y, ITH4012 also reduced amyloid beta25-35-induced apoptosis, determined by flow cytometry. ITH4012 caused a slight elevation in the cytosolic concentration of Ca2+ in fura 2-loaded bovine chromaffin cells, which could be related to the induction of protein synthesis relevant for cell survival. Blockade of protein synthesis by cycloheximide or blockade of Bcl-2's active site with HA14-1 (ethyl 2-amino-6-bromo-4-(1-cyano-2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate) reversed the cytoprotective action of ITH4012. Furthermore, exposure of bovine chromaffin cells for 24 or 48 h to neuroprotective concentrations of this compound enhanced, nearly 3-fold, the expression of the antiapoptotic protein Bcl 2. In conclusion, ITH4012 is a tacrine derivative that maintains acetylcholinesterase-inhibiting activity (IC50=0.8 microM) but has the additional property of acting as a calcium promotor, a property leading to neuroprotection through the induction of antiapoptotic proteins. PMID- 15111642 TI - BK channel news: full coverage on the calcium bowl. PMID- 15111643 TI - Mapping the BKCa channel's "Ca2+ bowl": side-chains essential for Ca2+ sensing. AB - There is controversy over whether Ca(2+) binds to the BK(Ca) channel's intracellular domain or its integral-membrane domain and over whether or not mutations that reduce the channel's Ca(2+) sensitivity act at the point of Ca(2+) coordination. One region in the intracellular domain that has been implicated in Ca(2+) sensing is the "Ca(2+) bowl". This region contains many acidic residues, and large Ca(2+)-bowl mutations eliminate Ca(2+) sensing through what appears to be one type of high-affinity Ca(2+)-binding site. Here, through site-directed mutagenesis we have mapped the residues in the Ca(2+) bowl that are most important for Ca(2+) sensing. We find acidic residues, D898 and D900, to be essential, and we find them essential as well for Ca(2+) binding to a fusion protein that contains a portion of the BK(Ca) channel's intracellular domain. Thus, much of our data supports the conclusion that Ca(2+) binds to the BK(Ca) channel's intracellular domain, and they define the Ca(2+) bowl's essential Ca(2+)-sensing motif. Overall, however, we have found that the relationship between mutations that disrupt Ca(2+) sensing and those that disrupt Ca(2+) binding is not as strong as we had expected, a result that raises the possibility that, when examined by gel-overlay, the Ca(2+) bowl may be in a nonnative conformation. PMID- 15111644 TI - Calcium regulation of single ryanodine receptor channel gating analyzed using HMM/MCMC statistical methods. AB - Type-II ryanodine receptor channels (RYRs) play a fundamental role in intracellular Ca(2+) dynamics in heart. The processes of activation, inactivation, and regulation of these channels have been the subject of intensive research and the focus of recent debates. Typically, approaches to understand these processes involve statistical analysis of single RYRs, involving signal restoration, model estimation, and selection. These tasks are usually performed by following rather phenomenological criteria that turn models into self fulfilling prophecies. Here, a thorough statistical treatment is applied by modeling single RYRs using aggregated hidden Markov models. Inferences are made using Bayesian statistics and stochastic search methods known as Markov chain Monte Carlo. These methods allow extension of the temporal resolution of the analysis far beyond the limits of previous approaches and provide a direct measure of the uncertainties associated with every estimation step, together with a direct assessment of why and where a particular model fails. Analyses of single RYRs at several Ca(2+) concentrations are made by considering 16 models, some of them previously reported in the literature. Results clearly show that single RYRs have Ca(2+)-dependent gating modes. Moreover, our results demonstrate that single RYRs responding to a sudden change in Ca(2+) display adaptation kinetics. Interestingly, best ranked models predict microscopic reversibility when monovalent cations are used as the main permeating species. Finally, the extended bandwidth revealed the existence of novel fast buzz-mode at low Ca(2+) concentrations. PMID- 15111645 TI - Modulation of the voltage sensor of L-type Ca2+ channels by intracellular Ca2+. AB - Both intracellular calcium and transmembrane voltage cause inactivation, or spontaneous closure, of L-type (CaV1.2) calcium channels. Here we show that long lasting elevations of intracellular calcium to the concentrations that are expected to be near an open channel (>/=100 microM) completely and reversibly blocked calcium current through L-type channels. Although charge movements associated with the opening (ON) motion of the channel's voltage sensor were not altered by high calcium, the closing (OFF) transition was impeded. In two-pulse experiments, the blockade of calcium current and the reduction of gating charge movements available for the second pulse developed in parallel during calcium load. The effect depended steeply on voltage and occurred only after a third of the total gating charge had moved. Based on that, we conclude that the calcium binding site is located either in the channel's central cavity behind the voltage dependent gate, or it is formed de novo during depolarization through voltage dependent rearrangements just preceding the opening of the gate. The reduction of the OFF charge was due to the negative shift in the voltage dependence of charge movement, as previously observed for voltage-dependent inactivation. Elevation of intracellular calcium concentration from approximately 0.1 to 100-300 microM sped up the conversion of the gating charge into the negatively distributed mode 10 100-fold. Since the "IQ-AA" mutant with disabled calcium/calmodulin regulation of inactivation was affected by intracellular calcium similarly to the wild-type, calcium/calmodulin binding to the "IQ" motif apparently is not involved in the observed changes of voltage-dependent gating. Although calcium influx through the wild-type open channels does not cause a detectable negative shift in the voltage dependence of their charge movement, the shift was readily observable in the Delta1733 carboxyl terminus deletion mutant, which produces fewer nonconducting channels. We propose that the opening movement of the voltage sensor exposes a novel calcium binding site that mediates inactivation. PMID- 15111646 TI - G-protein-gated TRP-like cationic channel activated by muscarinic receptors: effect of potential on single-channel gating. AB - There is little information about the mechanisms by which G-protein-coupled receptors gate ion channels although many ionotropic receptors are well studied. We have investigated gating of the muscarinic cationic channel, which mediates the excitatory effect of acetylcholine in smooth muscles, and proposed a scheme consisting of four pairs of closed and open states. Channel kinetics appeared to be the same in cell-attached or outside-out patches whether the channel was activated by carbachol application or by intracellular dialysis with GTPgammaS. Since in the latter case G-proteins are permanently active, it is concluded that the cationic channel is the major determinant of its own gating, similarly to the K(ACh) channel (Ivanova-Nikolova, T.T., and G.E. Breitwieser. 1997. J. Gen. Physiol. 109:245-253). Analysis of adjacent-state dwell times revealed connections between the states that showed features conserved among many other ligand-gated ion channels (e.g., nAChR, BK(Ca) channel). Open probability (P(O)) of the cationic channel was increased by membrane depolarization consistent with the prominent U-shaped I-V relationship of the muscarinic whole-cell current at negative potentials. Membrane potential affected transitions within each closed open state pair but had little effect on transitions between pairs; thus, the latter are likely to be caused by interactions of the channel with its ligands, e.g., Ca(2+) and Galphao-GTP. Channel activity was highly heterogeneous, as was evident from the prominent cycling behavior when P(O) was measured over 5-s intervals. This was related to the variable frequency of openings (as in the K(ACh) channel) and, especially, to the number of long openings between consecutive long shuttings. Analysis of the underlying Markov chain in terms of probabilities allowed us to evaluate the contribution of each open state to the integral current (from shortest to longest open state: 0.1, 3, 24, and 73%) as P(O) increased 525-fold in three stages. PMID- 15111648 TI - Historical perspective. PMID- 15111649 TI - Risk factors for TIA and TIA as a risk factor for stroke. PMID- 15111647 TI - Regulation of sodium channel function by bilayer elasticity: the importance of hydrophobic coupling. Effects of Micelle-forming amphiphiles and cholesterol. AB - Membrane proteins are regulated by the lipid bilayer composition. Specific lipid protein interactions rarely are involved, which suggests that the regulation is due to changes in some general bilayer property (or properties). The hydrophobic coupling between a membrane-spanning protein and the surrounding bilayer means that protein conformational changes may be associated with a reversible, local bilayer deformation. Lipid bilayers are elastic bodies, and the energetic cost of the bilayer deformation contributes to the total energetic cost of the protein conformational change. The energetics and kinetics of the protein conformational changes therefore will be regulated by the bilayer elasticity, which is determined by the lipid composition. This hydrophobic coupling mechanism has been studied extensively in gramicidin channels, where the channel-bilayer hydrophobic interactions link a "conformational" change (the monomer<-->dimer transition) to an elastic bilayer deformation. Gramicidin channels thus are regulated by the lipid bilayer elastic properties (thickness, monolayer equilibrium curvature, and compression and bending moduli). To investigate whether this hydrophobic coupling mechanism could be a general mechanism regulating membrane protein function, we examined whether voltage-dependent skeletal-muscle sodium channels, expressed in HEK293 cells, are regulated by bilayer elasticity, as monitored using gramicidin A (gA) channels. Nonphysiological amphiphiles (beta-octyl-glucoside, Genapol X 100, Triton X-100, and reduced Triton X-100) that make lipid bilayers less "stiff", as measured using gA channels, shift the voltage dependence of sodium channel inactivation toward more hyperpolarized potentials. At low amphiphile concentration, the magnitude of the shift is linearly correlated to the change in gA channel lifetime. Cholesterol-depletion, which also reduces bilayer stiffness, causes a similar shift in sodium channel inactivation. These results provide strong support for the notion that bilayer-protein hydrophobic coupling allows the bilayer elastic properties to regulate membrane protein function. PMID- 15111650 TI - TIA incidence and prevalence: the Stroke Belt perspective. PMID- 15111651 TI - TIAs and the pathology of cerebral ischemia. PMID- 15111652 TI - Reconsideration of TIA diagnostic criteria. PMID- 15111654 TI - A review of published TIA treatment recommendations. PMID- 15111653 TI - Neuroimaging in TIAs. PMID- 15111655 TI - Discussion: Reconsideration of TIA terminology and definitions. PMID- 15111656 TI - Carotid arteriography: still golden after all these years? PMID- 15111657 TI - A clever road from myopathology to genes: the myotilin story. PMID- 15111658 TI - ALS: persistent scientists do not find persisting enteroviruses. PMID- 15111659 TI - Efficacy and tolerability of the new antiepileptic drugs I: treatment of new onset epilepsy: report of the Therapeutics and Technology Assessment Subcommittee and Quality Standards Subcommittee of the American Academy of Neurology and the American Epilepsy Society. AB - OBJECTIVE: To assess the evidence demonstrating efficacy, tolerability, and safety of seven new antiepileptic drugs (AEDs) (gabapentin, lamotrigine, topiramate, tiagabine, oxcarbazepine, levetiracetam, and zonisamide-reviewed in the order in which these agents received approval by the US Food and Drug Administration) in the treatment of children and adults with newly diagnosed partial and generalized epilepsies. METHODS: A 23-member committee, including general neurologists, pediatric neurologists, epileptologists, and doctors in pharmacy, evaluated the available evidence based on a structured literature review including MEDLINE, Current Contents, and Cochrane library for relevant articles from 1987 until September 2002, with selected manual searches up until 2003. RESULTS: There is evidence either from comparative or dose-controlled trials that gabapentin, lamotrigine, topiramate, and oxcarbazepine have efficacy as monotherapy in newly diagnosed adolescents and adults with either partial or mixed seizure disorders. There is also evidence that lamotrigine is effective for newly diagnosed absence seizures in children. Evidence for effectiveness of the new AEDs in newly diagnosed patients with other generalized epilepsy syndromes is lacking. CONCLUSIONS: The results of this evidence-based assessment provide guidelines for the prescription of AEDs for patients with newly diagnosed epilepsy and identify those seizure types and syndromes where more evidence is necessary. PMID- 15111660 TI - Efficacy and tolerability of the new antiepileptic drugs II: treatment of refractory epilepsy: report of the Therapeutics and Technology Assessment Subcommittee and Quality Standards Subcommittee of the American Academy of Neurology and the American Epilepsy Society. AB - OBJECTIVE: To assess the evidence demonstrating efficacy, tolerability, and safety of seven new antiepileptic drugs (AEDs) (gabapentin, lamotrigine, topiramate, tiagabine, oxcarbazepine, levetiracetam, and zonisamide) in the treatment of children and adults with refractory partial and generalized epilepsies. METHODS: A 23-member committee including general neurologists, pediatric neurologists, epileptologists, and doctors in pharmacy evaluated the available evidence based on a structured literature review including MEDLINE, Current Contents, and Cochrane library for relevant articles from 1987 until March 2003. RESULTS: All of the new AEDs were found to be appropriate for adjunctive treatment of refractory partial seizures in adults. Gabapentin can be effective for the treatment of mixed seizure disorders, and gabapentin, lamotrigine, oxcarbazepine, and topiramate for the treatment of refractory partial seizures in children. Limited evidence suggests that lamotrigine and topiramate are also effective for adjunctive treatment of idiopathic generalized epilepsy in adults and children, as well as treatment of the Lennox Gastaut syndrome. CONCLUSIONS: The choice of AED depends upon seizure and/or syndrome type, patient age, concomitant medications, AED tolerability, safety, and efficacy. The results of this evidence-based assessment provide guidelines for the prescription of AEDs for patients with refractory epilepsy and identify those seizure types and syndromes where more evidence is necessary. PMID- 15111662 TI - Gene therapy of the brain: the trans-vascular approach. AB - Many chronic neurologic diseases do not respond to small molecule therapeutics, and have no effective long-term therapy. Gene therapy offers the promise of an effective cure for both genetic and acquired brain disease. However, the limiting problem in brain gene therapy is delivery to brain followed by regulation of the expression of the transgene. Present day gene vectors do not cross the blood brain barrier (BBB). Consequently, brain gene therapy requires craniotomy and the local injection of a viral gene vector. However, there are few brain disorders that can be effectively treated with local injection. Most applications of gene therapy require global expression in the brain of the exogenous gene, and this can only be achieved with a noninvasive delivery through the BBB--the trans vascular route to brain. An additional consideration is the potential toxicity of all viral and nonviral approaches, which may either integrate into the host genome and cause insertional mutagenesis or cause inflammation in the brain. Nonviral, noninvasive gene therapy of the brain is now possible with the development of a new approach to targeting therapeutic genes to the brain following an IV administration. This approach utilizes genetically engineered molecular Trojan horses, which ferry the gene across the BBB and into neurons. Global and reversible expression of therapeutic genes in the human brain without surgery and without viral vectors is now possible. PMID- 15111663 TI - Contrast-enhanced MR angiography for carotid disease: diagnostic and potential clinical impact. AB - OBJECTIVE: To compare contrast-enhanced MR angiography (CEMRA) with intra arterial digital subtraction angiography (DSA) for evaluating carotid stenosis. METHODS: A total of 167 consecutive symptomatic patients, scheduled for DSA following screening duplex ultrasound (DUS), were prospectively recruited to have CEMRA. Three independent readers reported on each examination in a blinded and random manner. Agreement was assessed using the Bland-Altman method. Diagnostic and potential clinical impact of CEMRA was evaluated, singly and in combination with DUS. RESULTS: CEMRA tended to overestimate stenosis by a mean bias ranging from 2.4 to 3.8%. A significant part of the disagreement between CEMRA and DSA was directly caused by interobserver variability. For detection of severe stenosis, CEMRA alone had a sensitivity of 93.0% and specificity of 80.6%, with a diagnostic misclassification rate of 15.0% (n = 30). More importantly, clinical decision-making would, however, have been potentially altered only in 6.0% of cases (n = 12). The combination of concordant DUS and CEMRA reduced diagnostic misclassification rate to 10.1% (n = 19) at the expense of 47 (24.9%) discordant cases needing to proceed to DSA. An intermediate approach of selective DUS review resulted in a marginally worse diagnostic misclassification rate of 11.6% (n = 22) but with only 6.8% of discordant cases (n = 13). CONCLUSIONS: DSA remains the gold standard for carotid imaging. The clinical misclassification rate with CEMRA, however, is acceptably low to support its safe use instead of DSA. The appropriateness of combination strategies depends on institutional choice and cost-effectiveness issues. PMID- 15111664 TI - Infarct patterns in atherosclerotic middle cerebral artery versus internal carotid artery disease. AB - OBJECTIVE: To compare clinical and radiologic characteristics of atherosclerotic middle cerebral artery (MCA) vs internal carotid artery (ICA) disease. METHODS: The authors defined atherosclerotic MCA and ICA disease as >50% symptomatic stenosis or occlusion without significant ICA and MCA stenosis on MR angiography. Patients with potential cardiac sources of embolism were excluded. The authors analyzed clinical, laboratory, and neuroradiologic data of the two groups. RESULTS: Among the 920 consecutive patients with acute ischemic strokes, 112 met the criteria for atherosclerotic MCA and 71 met the criteria for ICA disease. Clinically, the MCA group more frequently presented with lacunar syndrome (p = 0.001), whereas the ICA group more often presented with total anterior circulation infarct and had higher initial NIH Stroke Scale scores than the MCA group (all p < 0.001). Whereas deep perforator and internal border-zone infarcts were associated with MCA disease (p < 0.001 and 0.012), territorial infarcts and superficial perforator infarcts were associated with ICA disease (p < 0.001 and p = 0.009). The topographic patterns with respect to the degree of stenosis were also significantly different between the two groups. CONCLUSIONS: The clinical and radiologic stroke patterns were distinctively different between atherosclerotic MCA and ICA disease, suggesting different underlying pathogeneses. PMID- 15111665 TI - Cerebral lactic acidosis correlates with neurological impairment in MELAS. AB - OBJECTIVE: To evaluate the role of chronic cerebral lactic acidosis in mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS). METHODS: The authors studied 91 individuals from 34 families with MELAS and the A3243G point mutation and 15 individuals from two families with myoclonus epilepsy and ragged red fibers (MERRF) and the A8344G mutation. Subjects were divided into four groups. Paternal relatives were studied as controls (Group 1). The maternally related subjects were divided clinically into three groups: asymptomatic (no clinical evidence of neurologic disease) (Group 2), oligosymptomatic (neurologic symptoms but without the full clinical picture of MELAS or MERRF) (Group 3), and symptomatic (fulfilling MELAS or MERRF criteria) (Group 4). The authors performed a standardized neurologic examination, neuropsychological testing, MRS, and leukocyte DNA analysis in all subjects. RESULTS: The symptomatic and oligosymptomatic MELAS subjects had significantly higher ventricular lactate than the other groups. There was a significant correlation between degree of neuropsychological and neurologic impairment and cerebral lactic acidosis as estimated by ventricular MRS lactate levels. CONCLUSIONS: High levels of ventricular lactate, the brain spectroscopic signature of MELAS, are associated with more severe neurologic impairment. PMID- 15111666 TI - Glial and neuronal proteins in serum predict outcome after severe traumatic brain injury. AB - OBJECTIVE: To study the ability of glial (glial fibrillary acidic protein [GFAP] and S100b) and neuronal (neuron specific enolase [NSE]) protein levels in peripheral blood to predict outcome after severe traumatic brain injury. METHODS: Eighty-five patients with severe traumatic brain injury (admission Glasgow Coma Score [GCS] < or = 8) were included. Blood samples taken at the time of hospital admission were analyzed for S100b, GFAP, and NSE. Data collected included demographic and clinical variables. Outcome was assessed using the Glasgow Outcome Scale (GOS) at 6 months post injury. RESULTS: The median serum levels of S100b, GFAP, and NSE were raised 18.3 fold (S100b), 4.6 fold (GFAP), and twofold (NSE) compared to normal reference values. S100b, GFAP, and NSE serum levels correlated significantly with the injury severity score and CT findings but not with age, sex, or GCS. S100b, GFAP, and NSE levels were significantly higher in patients who died or had a poor outcome 6 months post injury than in those who were alive or had good outcome. S100b level >1.13 microg/L was the strongest predictor of death with 100% discrimination, but GFAP (>1.5 microg/L) and NSE (>21.7 microg/L) levels also strongly predicted death (adjusted odds ratios 5.82 [for GFAP] and 3.91 [for NSE]). S100b, GFAP, and NSE all strongly predicted poor outcome (adjusted odds ratios 5.12 [S100b], 8.82 [GFAP], and 3.95 [NSE]). CONCLUSIONS: These results suggest that determination of serum levels of glial and neuronal proteins may add to the clinical assessment of the primary damage and prediction of outcome after severe traumatic brain injury. PMID- 15111667 TI - Long-term sequelae in children after cerebellar astrocytoma surgery. AB - OBJECTIVE: To study long-term effects on neurologic, neuropsychological, and behavioral functioning in children treated for cerebellar pilocytic astrocytoma (CPA) without additional radio- and chemotherapy. METHODS: The authors assessed speech, language, nonverbal intelligence, attention, memory, executive skills, and visual (-spatial) functions in a consecutive series of 23 children. Neurologic and neuropsychological follow-up ranged from 1 year to 8 years and 10 months after resection. RESULTS: Long-term sequelae in the investigated domains were found in all children. Apraxia, motor neglect, and dysarthric features, as well as language, sustained attention, visual-spatial, executive, memory, and behavioral problems, were observed in various combinations and to different degrees. No clear pattern of neurocognitive disturbances could be discerned in this group. In addition, significant relationships were revealed between severity of preoperative hydrocephalus and visual-spatial skills. The high percentage of children who needed special education reflects the severity of the impairments. CONCLUSION: Despite the current opinion of a good quality of life after CPA treatment, careful long-term neurocognitive follow-up is needed in order to inform parents and teachers about the behavioral and cognitive sequelae and to contribute to timely social and educational intervention. PMID- 15111668 TI - Evaluation of visual recognition memory in MCI patients. AB - BACKGROUND: Neurofibrillary tangles seen early in Alzheimer disease (AD) initially appear in a subregion of the perirhinal cortex. In the monkey, damage to the perirhinal cortex impairs performance on visual recognition memory tasks. The authors evaluated impairment of visual recognition memory as a potential early diagnostic marker of AD. METHODS: The authors developed a visual delayed matching-to-sample task (DMS48) designed to assess visual recognition memory in humans. Twenty-three patients fulfilling the criteria of amnestic mild cognitive impairment (MCI) (mean Mini-Mental State Examination [MMSE]: 26.6, SD = 1.6) were recruited. All underwent a full neuropsychological evaluation, which included the Free and Cued Selective Reminding (FCSR) test. Their performance was compared with that of 10 patients with mild AD, 20 patients with moderate AD, 20 patients with Parkinson disease (PD), and 40 age-matched controls. RESULTS: Control subjects and patients with PD performed close to ceiling. Patients with mild AD had very low scores, while patients with moderate AD answered at random. MCI patients obtained scores that were between those of control subjects and patients with mild AD (78%, SD = 16%). MCI patients who failed on the DMS48 had lower scores on free recall (p < 0.05) and received less benefit from cueing (p < 0.01) on the FCSR than the other MCI, suggesting a profile of genuine memory impairment related to medial temporal lobe lesions. CONCLUSION: The DMS48, a test of visual recognition memory, is impaired early in the course of patients with MCI. Further studies are necessary to determine whether the evaluation of visual recognition memory may contribute to the identification of patients with AD. PMID- 15111669 TI - Role of the healthy hemisphere in recovery after resection of the supplementary motor area. AB - OBJECTIVE: To determine the compensatory mechanisms involved in the recovery of motor function following surgical lesions of the supplementary motor area (SMA) and their relation to the clinical characteristics of recovery. SUBJECTS AND METHODS: Twelve patients were referred for surgery of low-grade gliomas located in the SMA, and compared to eight healthy controls using fMRI before and after surgery during self-paced movements of both hands, successively. Magnitude and volume of activation within regions of interest (primary sensorimotor cortex, premotor cortex, SMA, preSMA, and parietal lobes) were compared and tested for correlation with anatomic characteristics of the tumor and resection, and clinical data. RESULTS: Tumor growth induced preoperative underactivity in the adjacent SMA and overactivity in the opposite SMA. Postoperative recovery was associated with recruitment of a premotor network located in the healthy hemisphere including the SMA and the lateral premotor cortex. Postoperative premotor recruitment in the healthy hemisphere increased with the percentage of resection of preoperative SMA activation. Shortened onset and duration of recovery was associated with increased preoperative changes in activation levels. CONCLUSIONS: These findings suggest a dysfunction of the SMA ipsilateral to the tumor, partially compensated by a recruitment of the contralesional SMA which correlated with shortened postoperative recovery. SMA resection was compensated by the recruitment of a medial and lateral premotor circuitry in the healthy hemisphere. PMID- 15111670 TI - Attentional bias induced by viewing actual or illusory movements. AB - OBJECTIVE: To learn to which portions of a line normal subjects would attend when watching this line actually moving or when perceiving movement even in the absence of actual movement, an illusory movement. METHODS: Twenty normal subjects watched a computer monitor on which either lines or dots moved to the right or left. They also watched stationary lines, which appeared to be moving to the left or right because the background was moving in the opposite direction. While watching these lines or dots, their eye movements were monitored to determine the position of fixation. RESULTS: Overall, subjects fixated on the side of the line that was in the direction of real or illusory movement. In the actual movement condition, leftward motion induced more of an attentional bias than rightward motion. In the illusory movement condition, however, rightward illusory movement condition induced more of an attentional bias than leftward movement. CONCLUSIONS: Objects moving leftward or rightward primarily activate the contralateral hemisphere. This hemispheric activation may induce a contralateral overt orienting response that is reflected by eccentric eye fixation. Treatments of neglect, such as caloric vestibular stimulation, may alter an attentional bias by inducing the illusion of movement. PMID- 15111671 TI - Chronic daily headache with analgesic overuse: epidemiology and impact on quality of life. AB - OBJECTIVE: To analyze the prevalence and demography of chronic daily headache (CDH) with analgesic overuse. METHODS: A population of 9,984 inhabitants aged 14 or older living in Santona, Spain, was studied. The authors personally interviewed 4,855 subjects, using a quota sampling approach. Those with headache for > or = 10 days/month and some analgesic use were asked to fill in a diary over the course of 1 month. Then, subjects were classified into CDH with or without analgesic overuse subtypes. Quality of life (Short Form-36 Health Survey [SF-36]) was also assessed in this second interview. RESULTS: Headache for > or = 10 days/month with analgesic consumption was reported by 332 subjects. Seven had secondary headache. Seventy-four (standardized prevalence 1.41%, 95% CI 1.1 to 1.8) fulfilled criteria for CDH with analgesic overuse. Prevalence in women (2.6%, 2.0 to 3.3) was much higher than in men (0.19%, 0.006 to 0.52). Mean age was 56 years (range 19 to 82 years). As recalled by the subjects, the mean age at onset of CDH was 38 years (range 9 to 82 years), whereas the mean age at onset of CDH with frequent analgesic consumption was 45 years (range 19 to 80 years) and that of primary headache was 22 years (range 5 to 60 years). CDH subjects showed a significant decrease in each SF-36 health-related score as compared with healthy control subjects. Transformed migraine was diagnosed in 49 (prevalence 0.9%), chronic tension-type headache in 20 (0.4%), and new daily persistent headache in 5 (0.1%). Thirty-five percent of patients overused simple analgesics, 22% ergotics, 12.5% opioids, and 2.7% triptans; the remaining 27.8% were overusing different combinations. CONCLUSION: CDH with analgesic overuse is a common disorder in the general population, mainly in women in their fifties, in whom 5% meet its diagnostic criteria. PMID- 15111672 TI - Limbic reductions of 5-HT1A receptor binding in human temporal lobe epilepsy. AB - OBJECTIVE: To test the hypothesis that in mesial temporal lobe epilepsy (MTLE) there is involvement outside of mesial structures and that this involvement affects serotonin systems, thus suggesting a mechanism for affective symptoms in this population. METHODS: Serotonin 5-HT1A receptor binding was studied with PET and [Carbonyl-11C]WAY-100 635 in 14 patients (6 with left-, 8 with right-sided mesial temporal lobe focus) and 14 controls. The 5-HT1A receptor binding potential was calculated for hippocampus, amygdala, orbitofrontal, insular, lateral temporal, and anterior cingulate cortex, in raphe nuclei, and in two regions presumably uninvolved in the epileptogenic process (parietal, and dorsolateral frontal neocortex). RESULTS: The binding potential was reduced in the epileptogenic hippocampus (p = 0.0001) and amygdala (p = 0.0001) in all patients, including the six with normal [18F]FDG PET and MRI. It was also reduced in the anterior cingulate (p = 0.002), insular (p = 0.015), and lateral temporal cortex (p = 0.029) ipsilaterally to the focus, in contralateral hippocampus (p = 0.025), and in the raphe nuclei (p = 0.016). CONCLUSION: Patients with severe MTLE show reduced 5-HT1A receptor binding potential in the EEG-focus, and its limbic connections. [(11)C]WAY-100 635 PET may provide additional information to EEG, [18F]FDG PET, and MRI when evaluating patients with intractable seizures. Reductions in 5-HT1A binding in the insula and cingulate suggest a mechanism by which affective symptoms in MTLE may result. PMID- 15111673 TI - Eclampsia, hippocampal sclerosis, and temporal lobe epilepsy: accident or association? AB - BACKGROUND: Up to 30% of patients with temporal lobe epilepsy (TLE) have no identifiable risk factors. OBJECTIVE: S: To report nine patients with TLE who had a history of eclampsia as the only risk factor for epilepsy and to investigate whether this possible association existed in a larger cohort of women with surgically treated TLE. METHODS: The clinical data, video-EEG, neuroimaging, and neuropathology of 195 consecutive women undergoing anterior temporal lobectomy (ATL) were reviewed. Risk factors for TLE, age at epilepsy onset, and occurrence of pregnancy were identified in each patient. RESULTS: Twenty-six women had no identifiable risk factors or seizures following a pregnancy. Nine of the 26 women had a history of eclampsia. The median age at the time of eclampsia was 16 years, and the latent period between the occurrence of eclampsia and onset of epilepsy ranged from 1 month to 2 years. The clinical, EEG, MRI, and neuropathologic findings were typical of hippocampal sclerosis (HS) and other than age at onset were no different from those of noneclampsia ATL patients. At mean follow-up of 57 months, seven patients were seizure-free and the other two markedly improved. CONCLUSION: Eclampsia may be a risk factor for TLE and HS. PMID- 15111674 TI - Stiff-person syndromes: motor cortex hyperexcitability correlates with anti-GAD autoimmunity. AB - OBJECTIVE: S: To investigate whether motor cortex excitability is enhanced in both stiff-person syndrome (SPS) and its "plus" variant, progressive encephalomyelitis with rigidity (PER), and related to autoimmunity against glutamic acid decarboxylase (GAD). METHODS: The authors compared 21 patients with SPS or PER (7 untreated, 14 treated) with 14 age-matched healthy controls and used transcranial magnetic stimulation (TMS, paired-pulse paradigm) to investigate intracortical inhibition (ICI) and intracortical facilitation (ICF). GAD autoantibody levels in serum and CSF were determined by radioimmunoassay. RESULTS: The authors found significantly enhanced motor cortex excitability in untreated SPS and PER patients. GABAmimetic medication significantly reduced ICF but did not affect ICI. Motor cortex excitability was more enhanced in patients with GAD antibodies than in patients without GAD antibodies and correlated positively with GAD antibody levels in CSF. CONCLUSIONS: The motor cortex is hyperexcitable in SPS and PER patients. However, hyperexcitability is partly masked by GABAmimetic treatment. Correlation of elevated GAD antibody levels with enhanced ICF suggests that motor cortex hyperexcitability in SPS and PER is related to anti-GAD autoimmunity. PMID- 15111675 TI - Mutations in myotilin cause myofibrillar myopathy. AB - BACKGROUND AND OBJECTIVE: The term myofibrillar myopathy (MFM) is a noncommittal term for a pathologic pattern of myofibrillar dissolution associated with accumulation of myofibrillar degradation products and ectopic expression of multiple proteins. Ultrastructural studies implicate the Z-disk as the site of the initial pathologic change, and mutations in two Z-disk-related proteins, desmin and alphaB-crystallin, have been identified in a minority of patients with MFM. The authors' objective was to determine whether mutations in myotilin, a key Z-disk component and the disease protein in limb-girdle muscular dystrophy (LGMD) 1A, are another cause of MFM. METHODS: The authors used histochemical, immunocytochemical, ultrastructural, and mutation analysis. RESULTS: The authors detected four missense mutations in 6 of 57 patients with MFM in the serine-rich exon 2 of MYOT, where the two previously identified LGMD1A mutations are located. Three mutations were novel, and one had been previously identified in LGMD1A. Each patient had evidence for neuropathy, and at least three kinships had associated cardiomyopathy. Distal weakness greater than proximal weakness was present in three patients. Except for minor differences, the morphologic features were similar to those in other patients with MFM. CONCLUSIONS: 1) Mutations in myotilin cause MFM; 2) exon 2 of MYOT is a hotspot for mutations; 3) peripheral neuropathy, cardiomyopathy, and distal weakness greater than proximal weakness are part of the spectrum of myotilinopathy; 4) not all cases of myotilinopathy have a limb-girdle phenotype; and 5) the molecular basis of the majority of MFM cases remains to be discovered. PMID- 15111676 TI - Failure to detect enterovirus in the spinal cord of ALS patients using a sensitive RT-PCR method. AB - OBJECTIVE: To assess the association of enteroviruses (EV) with ALS by applying a sensitive seminested reverse transcription (RT) PCR protocol to the detection of enteroviral RNA in a blinded set of archived tissues from ALS and control cases. METHODS: The specimen set consisted of 24 frozen spinal cord samples from ALS cases, 17 frozen spinal cord samples from negative control (non-ALS) cases, and 5 frozen spinal cord positive control samples. The positive controls were two human spinal cord samples spiked with poliovirus (PV) and three spinal cords from PV infected transgenic mice. A sensitive, EV-specific, seminested RT-PCR assay was used to detect EV genome in RNA extracted from the specimens and controls. RESULTS: The assay detected EV RNA in a 10(-5) dilution of infected mouse tissue. EV RNA was not detected in the ALS specimens or in specimens from control cases, despite the presence of amplifiable RNA as assessed by amplification with control primers, whereas all of the positive control specimens yielded the expected PV amplification product. CONCLUSION: The reported association between EV infection and ALS was not confirmed by testing this set of specimens with these sensitive methods. PMID- 15111677 TI - Symptomatic distal sensory polyneuropathy in HIV after age 50. AB - OBJECTIVE: To determine if aging changes the frequency, severity, or manifestations of symptomatic distal sensory polyneuropathy (SxDSPN) in patients with HIV-1. METHODS: Prospective observations of 70 older (age < or = 50) and 56 younger (age 20 to 40) patients with HIV, and a control group of 48 older non-HIV patients, were conducted utilizing neurologic examination, neuropsychological testing, lumbar puncture, laboratory, and medical history. RESULTS: The frequency of SxDSPN among older HIV patients was 50.4%, compared to 19.6% among younger HIV patients (p < 0.001). SxDSPN among control patients occurred in 4.2%, similar to the general population. Older compared to younger HIV patients demonstrated more severe symptoms (p = 0.02) and greater deficits for vibration (p < 0.01). Increasing numbers of neuropathic comorbidities among older compared to younger HIV patients were associated with increasing severity of deficits to pinprick (p = 0.003). Dementia and SxDSPN coexisted in 36% of the older HIV patients and in none of the younger HIV patients (p = 0.021). Older HIV patients with nadir CD4 < or =200 cells/mL were 4.23 times as likely to have SxDSPN than older patients with nadir CD4 >200 cells/mL (p = 0.007). Vibratory deficits excessive to pinprick deficits predicted SxDSPN among older (OR 2.83) but not younger seropositive patients (p = 0.036). CONCLUSIONS: Age > or = 50 increases the frequency of SxDSPN, and is associated with both vibratory loss as the predominant sensory deficit and increased severity of pinprick loss among symptomatic patients with neuropathic comorbidities. SxDSPN is associated with both dementia and low nadir CD4 in HIV-positive patients aged 50 and greater. PMID- 15111678 TI - Regional metabolism in primary torsion dystonia: effects of penetrance and genotype. AB - BACKGROUND: The authors have previously used [18F]fluorodeoxyglucose (FDG) PET to identify a reproducible pattern of regional glucose metabolism that was expressed in both manifesting and nonmanifesting carriers of the DYT1 primary dystonia mutation. OBJECTIVE: To identify specific regions that discriminated subjects according to clinical penetrance and genotype. METHODS: FDG PET was used to scan 12 nonmanifesting and 11 manifesting DYT1 gene carriers, 6 nonmanifesting DYT6 gene carriers and 7 manifesting DYT6 gene carriers, as well as 11 control subjects. The data from all five groups were analyzed with statistical parametric mapping and analysis of variance with posthoc contrasts. RESULTS: A dissociation of metabolic changes was found related to phenotype and genotype. Manifesting gene carriers of both genotypes exhibited bilateral hypermetabolism in the presupplementary motor area (Brodmann area [BA] 6) and parietal association cortices (BA 40/7) compared with the respective nonmanifesting counterparts. By contrast, genotype-specific increases in metabolism were found in the putamen, anterior cingulate (BA 24/32), and cerebellar hemispheres of DYT1 carriers. Genotype-specific changes in DYT6 involved hypometabolism of the putamen and hypermetabolism in the temporal cortex (BA 21). CONCLUSIONS: Dystonia may be associated with abnormal movement preparation caused by defective sensorimotor integration. Whereas clinical manifestations are related to cortical dysfunction, metabolic abnormalities in subcortical structures may represent trait features that are specific for individual dystonia genotypes. PMID- 15111679 TI - Efficacy of pergolide in treatment of restless legs syndrome: the PEARLS Study. AB - OBJECTIVE: To evaluate the short- and long-term safety and efficacy of pergolide therapy for restless legs syndrome (RLS) in a double-blind, placebo-controlled, randomized trial (Pergolide European Australian RLS [PEARLS] study). METHODS: We randomized 100 patients with idiopathic RLS were randomized to pergolide, 0.25 to 0.75 mg, in the evening or placebo for 6 weeks (phase 1); thereafter, patients with response on the Patient Global Impression (PGI) scale continued on double blind pergolide or placebo, and nonresponders received open-label pergolide up to 1.5 mg/d for 12 months of treatment (phase 2). Sleep efficiency (SE) and periodic limb movements during sleep (PLMS) arousal index were monitored by centrally evaluated polysomnography (PSG). The severity of RLS was assessed using the validated International RLS Scale (IRLS). RESULTS: In phase 1 (change from baseline to week 6), pergolide reduced PLMS arousal index vs placebo (mean +/- SD, -12.6 +/- 10.0 vs -3.6 +/- 15.9; p = 0.004), and SE did not improve (mean +/- SD, +11.3 +/- 11.9% vs +6.1 +/- 18.6%; p = 0.196). Pergolide improved RLS severity score (-12.2 +/- 9.9 vs -1.8 +/- 7.5 placebo; p < 0.001) and was associated with a higher PGI response (68.1% vs 15.1%; p < 0.001) and improvements in periodic limb movements (PLM) index, PGI improvement scale, Clinical Global Impression improvement, and IRLS (all p < 0.001), patient reported SE (p = 0.019), and quality of sleep (p < 0.001). After 12 months (phase 2), double-blind pergolide maintained improvements in PLMS arousal index and PLM index. Placebo patients switched to open-label pergolide in phase 2 exhibited marked improvements in these measures that were maintained at 12 months. Pooled results from the blinded and open-label pergolide groups demonstrated improvements at 12 months in the PLMS arousal index (p = 0.028) and PLM index (p < 0.0001) compared with placebo. Nausea and headache were more frequent with pergolide than with placebo treatment. CONCLUSIONS: Pergolide substantially improves periodic limb movement measures and subjective sleep disturbance associated with restless legs syndrome. Low-dose pergolide was well tolerated and maintained its efficacy in the long term. PMID- 15111680 TI - Hemodynamic changes in the basilar artery following stenting. PMID- 15111681 TI - Percutaneous closure of patent foramen ovale reduces the frequency of migraine attacks. AB - Among 215 patients referred for percutaneous closure of patent foramen ovale (PFO) after presumed paradoxical embolism, we assessed the prevalence of migraine. In the year prior to PFO closure, 48 (22%) patients had migraine, twice the expected prevalence of 10 to 12% in the general European population. In patients with migraine with aura, percutaneous PFO closure reduced the frequency of migraine attacks by 54% (1.2 +/- 0.8 vs 0.6 +/- 0.8 per month; p = 0.001) and in patients with migraine without aura by 62% (1.2 +/- 0.7 vs 0.4 +/- 0.4 per month; p = 0.006). PFO closure did not have an effect on headache frequency in patients with nonmigraine headaches (1.4 +/- 0.9 vs 1.0 +/- 0.9 per month; p = NS). PMID- 15111682 TI - Paraneoplastic "stiff person syndrome" with metastatic adenocarcinoma and anti-Ri antibodies. AB - A 43-year-old woman presented with clinical and electrophysiologic features of stiff person syndrome (SPS), without abdominal or lumbar paraspinal muscle involvement. Investigations revealed metastatic adenocarcinoma of the lung with positive anti-Ri antibodies. Her clinical condition improved with diazepam, baclofen, tizanidine, and palliative chemotherapy. Screening for an underlying malignancy and anti-Ri antibodies should be considered in patients with SPS when clinical presentation is atypical. PMID- 15111683 TI - Ropinirole for restless legs syndrome: a placebo-controlled crossover trial. AB - The authors performed a double-blind, placebo-controlled, crossover study of ropinirole (0.5 to 6.0 mg/day) for restless legs syndrome (RLS). The RLS Rating Scale score improved (p < 0.001) from a mean (SD) of 25 (7) during placebo treatment to 13 (12) during ropinirole treatment. Eight of the 22 patients had complete resolution of symptoms on ropinirole. Adverse events included nausea and dizziness. Ropinirole was effective and well tolerated for treating the symptoms of RLS. PMID- 15111684 TI - Myoclonic status epilepticus following repeated oral ingestion of colloidal silver. AB - The authors report a case of a 71-year-old man who developed myoclonic status epilepticus and coma after daily ingestion of colloidal silver for 4 months resulting in high levels of silver in plasma, erythrocytes, and CSF. Despite plasmapheresis, he remained in a persistent vegetative state until his death 5.5 months later. Silver products can cause irreversible neurologic toxicity associated with poor outcome. PMID- 15111685 TI - Hypnic headache: PSG evidence of both REM- and NREM-related attacks. AB - Hypnic headache (HH) occurs exclusively during sleep. Six attacks were recorded during nocturnal polysomnographic (PSG) monitoring of 10 HH patients. The PSG data obtained indicate that the attacks arose directly from sleep: four from non REM and two from REM sleep. In no patient were the HH attacks found to show any close temporal relationship with sleep-related breathing abnormalities. PMID- 15111686 TI - Nimodipine for treatment of primary thunderclap headache. AB - Eleven patients with primary thunderclap headache (TCH) were treated with oral nimodipine 30 to 60 mg every 4 hours or IV nimodipine 0.5 to 2 mg/h if the oral regimen failed or images showed cerebral vasospasm. With oral nimodipine, headache did not recur in the nine patients without vasospasm. IV nimodipine was given in two patients with vasospasm, including one who developed ischemic stroke. Nimodipine may be effective for TCH. Vasospasm may warrant IV nimodipine. PMID- 15111687 TI - Brainstem involvement in hypertensive encephalopathy: clinical and radiological findings. AB - Predominant brainstem or cerebellar edema is rare in hypertensive encephalopathy and usually affects patients with secondary hypertension. Despite the severity of the radiologic findings, clinical features of brainstem involvement are uncommon. The authors report the clinical and radiologic features of two patients. PMID- 15111688 TI - Catastrophic presentation of mitochondrial disease due to a mutation in the tRNA(His) gene. AB - The authors describe a patient who presented with headache, seizures, and severe cerebral edema in whom they identified a novel mutation in the mitochondrial (mt ) tRNA(His) gene. This G12147A transition is heteroplasmic, predicted to disrupt a highly conserved base pair, and segregates with the cytochrome c oxidase deficiency in single muscle fibers. PMID- 15111689 TI - Apparent weakness of ulnar-innervated muscles in radial palsy. AB - Muscle strength was assessed in 11 patients with radial or posterior interosseus palsy. Apparent weakness was found in the dorsal and palmar interossei and the abductor digiti minimi. These muscles insert on the extensor expansions, and their activation is associated with concomitant contraction of finger flexors and extensors. This apparent weakness may be due to their unopposed traction on the extensor expansion by the paralyzed extensor digitorum. PMID- 15111690 TI - Magnetic resonance imaging of thoracic epidural venous dilation in Hirayama disease. AB - Hirayama disease is a nonprogressive, asymmetric amyotrophy of the hands and forearms, possibly caused by compression of the lower cervical cord on neck flexion. The authors used phase-contrast MR angiography to study a patient with this disorder and observed abnormal spinal epidural venous dilation on neck flexion. In addition to mechanical compression of the lower cervical cord, venous congestion in the spinal canal may have a role in promoting anterior horn damage. PMID- 15111691 TI - Giant axon and neurofilament accumulation in Charcot-Marie-Tooth disease type 2E. AB - The axonal type 2 Charcot-Marie-Tooth disease (CMT2) is phenotypically poorly characterized. Here the authors report a family with a Pro22Ser mutation in the neurofilament-light gene (NF-L; CMT2E) manifesting electrophysiologically as the demyelinating type 1 CMT (CMT1) and pathologically as an axonopathy with giant axons and accumulation of disorganized NF. NF-L should be investigated in CMT2 as well as in CMT1 not associated with the usual genes PMP22, Cx32, and P0. PMID- 15111692 TI - Measurement error of two different techniques for brain atrophy assessment in multiple sclerosis. AB - The measurement errors associated with two techniques for brain atrophy assessment in multiple sclerosis (MS) were estimated. The measurement error of a semiautomated, nonnormalized technique with partial brain coverage was about twofold higher than that of a fully automated, normalized technique with full brain coverage (Structural Image Evaluation of Normalized Atrophy software [SIENA]). As shown by the analysis of data from a clinical trial of glatiramer acetate, the use of SIENA increases the study power to detect a treatment effect on brain volume changes in MS patients. PMID- 15111693 TI - Reversible parkinsonism with normal beta-CIT-SPECT in patients exposed to sodium valproate. AB - After reports of reversible parkinsonism and cognitive impairment with sodium valproate (VPA), the authors examined 50 consecutive patients taking VPA and 20 patients taking carbamazepine. Three patients taking VPA exhibited unequivocal parkinsonism with Unified Parkinson Disease Rating Scale scores >30. VPA was withdrawn from two patients with improvement of symptoms. Reduction in VPA dosage in the third patient produced no improvement. beta-CIT-SPECT scans were normal, suggesting dopaminergic neuronal loss is not the underlying mechanism. PMID- 15111694 TI - Cerebral air embolism during transbronchial biopsy. PMID- 15111695 TI - Closure of a patent foramen ovale is associated with a decrease in prevalence of migraine. PMID- 15111696 TI - Prolactinoma presenting as painful postganglionic Horner syndrome. PMID- 15111697 TI - IV insulin during acute cerebral infarction in diabetic patients. PMID- 15111698 TI - Sudden unilateral deafness due to a right vertebral artery dissection. PMID- 15111699 TI - Insulinoma misdiagnosed as intractable epilepsy. PMID- 15111700 TI - Plasmapheresis improves outcome in postinfectious cerebellitis induced by Epstein Barr virus. PMID- 15111701 TI - Changes in the appearance of venous sinuses after treatment of disordered intracranial pressure. PMID- 15111702 TI - Auditory dysfunction in chronic inflammatory demyelinating polyradiculoneuropathy. PMID- 15111703 TI - Memoir: Education of a gentile on the history of the Holocaust. PMID- 15111704 TI - Cardiac troponin levels following monitored epileptic seizures. PMID- 15111705 TI - Retrospective diagnosis of congenital cytomegalovirus infection and cortical maldevelopment. PMID- 15111706 TI - CSF hypovolemia vs intracranial hypotension in "spontaneous intracranial hypotension syndrome". PMID- 15111707 TI - L-dopa-induced excessive daytime sleepiness in PD: a placebo-controlled case with MSLT assessment. PMID- 15111708 TI - Severe restless legs syndrome presenting as intractable insomnia. PMID- 15111709 TI - Patient page. Does the percutaneous closure of patent foramen ovale help the migraine sufferer? PMID- 15111710 TI - The multiple-stress responsive plastid sigma factor, SIG5, directs activation of the psbD blue light-responsive promoter (BLRP) in Arabidopsis thaliana. AB - Transcription in higher plant plastids is performed by two types of RNA polymerases called NEP and PEP, and expression of photosynthesis genes in chloroplasts is largely dependent on PEP, a eubacteria-type multi-subunit enzyme. The transcription specificity of PEP is modulated by six nuclear-encoded sigma factors (SIG1 to SIG6) in Arabidopsis thaliana. Here, we show that one of the six sigma factors, SIG5, is induced under various stress conditions, such as high light, low temperature, high salt and high osmotic conditions. Interestingly, transcription from the psbD blue light-responsive promoter (psbD-BLRP) was activated by not only light but also various stresses, and the transcription and the transcriptional activation of psbD-BLRP were abolished in a sig5-2 mutant. This suggests that the PEP holoenzyme containing SIG5 transcribes the psbD-BLRP in response to multiple stresses. Since the seed germination under saline conditions and recovery from damage to the PSII induced by high light were delayed in the sig5-2 mutant, we postulate that SIG5 protects plants from stresses by enhancing repair of the PSII reaction center. PMID- 15111711 TI - Amphivasal vascular bundle 1, a gain-of-function mutation of the IFL1/REV gene, is associated with alterations in the polarity of leaves, stems and carpels. AB - In Arabidopsis stems, the vascular bundles in the stele are arranged in a ring like pattern and the vascular tissues in each bundle are organized in a collateral pattern. We have shown previously that the semidominant amphivasal vascular bundle 1 (avb1) mutation transforms the collateral vascular bundles into amphivasal bundles and disrupts the ring-like arrangement of vascular bundles in the stele. In this study, we show that the avb1 mutation occurred in the putative microRNA 165 target sequence in the IFL1/REV gene and caused an amino acid substitution in the putative sterol/lipid-binding START domain. We present direct evidence that the wild-type IFL1/REV mRNA was cleaved within the microRNA 165 target sequence and the avb1 mutation resulted in an inhibition of cleavage and a higher level accumulation of full-length mRNA, suggesting a role of microRNA 165 in the regulation of IFL1/REV gene expression. In addition to an alteration in vascular patterning, the avb1 mutation also caused dramatic changes in fiber cell wall thickening and organ polarity, including aberrant formation and proliferation of cauline leaves and branches, production of trumpet-shaped leaves with reversed adaxial-abaxial identity, ectopic growth of carpel-like structures on the outer surface of carpels, and fasciation of inflorescence. Ectopic overexpression of the avb1 mutant cDNA not only phenocopied most of the avb1 mutant phenotypes but also led to additional novel phenotypes such as formation of leaves with extremely narrow blades and ectopic production of branches in the axil of siliques. Taken together, these results suggest that the avb1 gain-of function mutation of the IFL1/REV gene alters the positional information that determines vascular patterning and organ polarity. PMID- 15111712 TI - Dof domain proteins: plant-specific transcription factors associated with diverse phenomena unique to plants. AB - Dof (DNA-binding with one finger) domain proteins are plant-specific transcription factors with a highly conserved DNA-binding domain, which presumably includes a single C(2)-C(2) zinc finger. During the past decade, numerous Dof domain proteins have been identified in both monocots and dicots including maize, barley, wheat, rice, tobacco, Arabidopsis, pumpkin, potato, and pea. Biochemical, molecular biological and molecular genetic analyses revealed that Dof domain proteins function as a transcriptional activator or a repressor involved in diverse plant-specific biological processes. Although more physiological roles of Dof domain proteins would be elucidated in future because of numerous Dof domain proteins in plants, it is already evident that the Dof domain proteins play critical roles as transcriptional regulators in plant growth and development. Here I summarize our current knowledge about Dof domain proteins. PMID- 15111713 TI - On the 710 nm fluorescence emitted by the diatom Phaeodactylum tricornutum at room temperature. AB - The fluorescence emitted at 710 nm by Phaeodactylum tricornutum (F(710)) was characterized. Development of F(710) was found to be regulated by the quality of light needed for algal growth: weak red light absorbed mainly by Chl a induced its development, and weak blue-green light absorbed mainly by fucoxanthin and Chl c suppressed it. The difference spectra between cells grown under the two light conditions revealed two Chl a forms, absorption peaks of which were located at 692 nm (Chl a(692)) and at 703 nm (Chl a(703)), respectively, in red-light-grown cells. During cell growth under red light, the appearance and intensification of the emission correlated well with development of Chl a(692) and Chl a(703) suggesting that the two forms of Chl a are involved in the energy flow to F(710). A clear induction phenomenon characteristic of the PSII fluorescence was observed not only with the emission at 680 nm but also with F(710), indicating that F(710) is emitted by PSII Chl a. Development of F(710) under red light was sensitive to cycloheximide, indicating that the development of the energy flow to F(710) requires protein synthesis and that the emitter is installed in a protein encoded in the nuclear genome like the light-harvesting complex (LHC). Centrifugal fractionation of pigment-protein complexes revealed F(710) to be located at fractions slightly heavier than the major LHC. Development of F(710) was also found in red-light-grown cells of the diatom Nitzschia closterium. PMID- 15111714 TI - Flowering of Arabidopsis cop1 mutants in darkness. AB - To elucidate the role of the COP1 gene in flowering, we analyzed flowering of cop1 mutant lines in darkness. When grown in the presence of 1% (w/v) sucrose, the cop1-6 mutant flowered in darkness, but cop1-1 and cop1-4 did not. However, cop1-1 and cop1-4 flowered in darkness when grown in the presence of 5% (w/v) sucrose. Therefore, the COP1 gene represses not only photomorphogenesis in seedlings but also flowering in darkness. Comparison of mRNAs levels of floral identity genes in cop1-6 and wild-type plants grown in darkness revealed increased mRNA levels of genes that act downstream of CO and reduced FLC mRNA level in cop1-6. Double mutants of cop1-6 and each of the late-flowering mutations cry2-1, gi-2, co-1, and ld-1 flowered in darkness. All of the double mutants except cry2-1 cop1-6 flowered later than cop1-6, demonstrating that cop1 6 is epistatic to cry2-1 for early flowering. The ld-1 cop1-6 double mutant flowered much earlier than the ld-1 mutant. The delay in flowering in the double mutants was not strongly influenced by the light conditions, whereas that of the gi-2 cop1-6 double mutant was reduced in darkness. PMID- 15111715 TI - Thioredoxin reduction alters the solubility of proteins of wheat starchy endosperm: an early event in cereal germination. AB - A KCl-soluble, albumin/globulin fraction of wheat (Triticum aestivum L.) starchy endosperm was further separated into a methanol-insoluble fraction that contained metabolic proteins and a methanol-soluble fraction that contained "chloroform methanol" or CM-like proteins. Reduction of the disulfide bonds of the CM proteins with thioredoxin or dithiothreitol altered their properties so that, like the metabolic proteins, they were insoluble in methanol. Glutathione had little effect, indicating dithiol specificity. Proteomic analysis of the CM protein fraction revealed the presence of isoforms of low molecular weight disulfide proteins (alpha-amylase, alpha-amylase/trypsin and WCI proteinase inhibitors, lipid transfer proteins, gamma-thionins), stress enzymes (Cu-Zn superoxide dismutase and peroxidase), storage proteins (alpha-, gamma- and omega gliadins, low molecular weight glutenin subunits and globulins of the avenin N9 type), and a component of protein degradation (polyubiquitin). These findings support the view that, in addition to modifying activity and increasing protease sensitivity, reduction by thioredoxin alters protein solubility, thereby promoting processes of the grain starchy endosperm, notably the mobilization of reserves during germination and seedling development. PMID- 15111716 TI - Function analysis of phototropin2 using fern mutants deficient in blue light induced chloroplast avoidance movement. AB - Gametophytes of the fern Adiantum capillus-veneris L. were mutagenized by heavy ion beam irradiation and screened for mutants lacking chloroplast avoidance movement under high intensity blue light. Mutants recovered include several with small deletions in the AcPHOT2 gene. The avoidance movement response in these mutants could be restored by transient expression of non-mutant AcPHOT2 cDNA, indicating that the chloroplast avoidance movement in this fern is mediated by the Acphot2 protein. Further functional analyses of the Acphot2 protein were performed using this transient assay for chloroplast avoidance movement. The results obtained suggest that the LOV2, but not the LOV1, domain of Acphot2 is essential for avoidance movement, and that several residues in the C-terminus of the kinase domain contribute to the avoidance response. The rate of dark reversion of the photo-activated LOV2 domain, which was calculated photometrically, was too fast to account for the lifetime of phot2 signal estimated from physiological responses. However, the rate of dark reversion of the combined domains of LOV1 and LOV2 did correspond to the lifetime of the signal, suggesting that LOV1 might have some function in this response, although it is not essential for playing a role as a photoreceptor. PMID- 15111717 TI - Transgenic Lotus japonicus with an ethylene receptor gene Cm-ERS1/H70A enhances formation of infection threads and nodule primordia. AB - Ethylene inhibits the establishment of symbiosis between rhizobia and legumes. To examine how and when endogenous ethylene inhibits rhizobial infection and nodulation, we produced transgenic Lotus japonicus carrying the mutated melon ethylene receptor gene Cm-ERS1/H70A that confers ethylene insensitivity and fixes the transgene in the T(3) generation. The resultant transgenic plants showed reduced ethylene sensitivity because of 1-aminocyclopropane-1-carboxylate resistance and increased flowering duration, probably due to a dominant negative mechanism. When inoculated with Mesorhizobium loti, transgenic plants showed markedly higher numbers of infection threads and nodule primordia on their roots than did either wild-type or azygous plants during the early stage of cultivation period as well as during later stages, when the number of mature nodules had reached a steady state. In addition, transcripts of NIN, a gene governing infection thread formation, increased in the inoculated transgenic plants as compared with the wild-type plants. The infection responses of transgenic plants were similar to those of wild-type plants treated with ethylene inhibitors. These results imply that the endogenous ethylene in L. japonicus roots inhibits rhizobial infection at the primary nodulation, probably via NIN gene, and suggest that ethylene perception assists negative feedback regulation of secondary nodule initiation. PMID- 15111718 TI - Molecular cloning of two exo-beta-glucanases and their in vivo substrates in the cell walls of lily pollen tubes. AB - Full-length cDNA sequences of two exo-beta-glucanases, LP-ExoI and LP-ExoII, secreted into cell walls of lily (Lilium longiflorum) pollen tube, were determined by RT-PCR. LP-ExoI exhibited over 80% similarity to LP-ExoII at both DNA and amino acid levels. RT-PCR showed that LP-ExoI transcripts were abundant in pollen grains and tubes, but could not be detected in leaf, stem, stigma, style, ovary, petal, filament, young root, young bud, and scale leaf of bulb. However, LP-ExoII transcripts ubiquitously existed in all the tissues tested. To determine the potential substrates of exo-beta-glucanases, cell wall components of lily tissues were analyzed. Linkage analysis revealed that pollen tubes contained high levels of 3-Glc in hemicellulose (44.3%), while pollen grains had no detectable 3-Glc. The hemicellulose fraction of pollen tubes was treated with lichenase and the product was analyzed by HPLC-PAD to determine the origin of 3 Glc. Specific tetra-saccharide was liberated from hemicellulose of pollen tubes, suggesting the presence of 1,3 : 1,4-beta-glucan in lily pollen tube hemicellulose. The structure of this 1,3 : 1,4-beta-glucan may be different from cereal plant 1,3 : 1,4-beta-glucan, since tri-saccharide was not detected in hemicellulose fraction after lichenase treatment. LP-ExoI and LP-ExoII, expressed in pollen grains and tubes, may be involved in the regulation of pollen tube elongation by hydrolyzing callose and 1,3 : 1,4-beta-glucan within pollen tube walls. PMID- 15111719 TI - A deficiency at the gene coding for zeta-carotene desaturase characterizes the sunflower non dormant-1 mutant. AB - The non dormant-1 (nd-1) mutant of sunflower (Helianthus annuus L.) is characterized by an albino and viviparous phenotype. Pigment analysis by spectrophotometer and HPLC demonstrated in nd-1 cotyledons the absence of beta carotene, lutein and violaxanthin. Additionally, we found a strong accumulation of zeta-carotene and, to a lesser extent, of phytofluene and cis-phytoene in nd-1 seedlings grown in very dim light (1 micro mol m(-2) s(-1)). These results suggested that zeta-carotene desaturation was impaired in the mutant plants. To understand the molecular basis of the nd-1 mutation, we cloned and characterized the zeta-carotene desaturase (Zds) gene from sunflower. A reconstructed full length sequence (1,916 bp) of the Zds cDNA was obtained from homozygous Nd-1/Nd-1 wild-type plants. It contains a 1,761-bp CDS, 62 nucleotides of 5'-untranslated region (UTR), and 77 nucleotides of 3'-UTR. The predicted protein (64.9 kDa) consists of 587 amino acid residues with a putative transit sequence for plastid targeting in the N-terminal region and a typical amino oxidase domain that includes the flavin adenosine dinucleotide (FAD) binding motif. The phylogenetic analysis demonstrated that the sunflower Zds was clustered to marigold (Tagetes) Zds gene, for which it showed an overall aminoacidic identity of 96.6% and resulted strictly correlated with other Zds sequences of higher plants. Interestingly, RT-PCR analyses showed that nd-1 plants were unable to accumulate Zds transcripts. Sequence information from the Zds cDNA was used to design specific primers and to isolate the full-length exons/introns region of the gene. The sunflower Zds gene (HaZds) comprises 14 exons and 13 introns scattered in a ca. 5.0-kb region. Also, HaZds showed a high conservation of the distribution and size of the exons with rice Zds gene. Based on genomic Southern analysis, the nd 1 genome disclosed a large deficiency at the Zds locus. PMID- 15111720 TI - Analysis of the transient increase in cytosolic Ca2+ during the action potential of higher plants with high temporal resolution: requirement of Ca2+ transients for induction of jasmonic acid biosynthesis and PINII gene expression. AB - Plants respond to various abiotic stimuli by activation and propagation of fast electrical signals, action potentials. To resolve the temporal increase in cytosolic Ca(2)(+) during the action potentials of higher plants, we regenerated transgenic potato plants that expressed the Ca(2)(+) photoprotein apoaequorin. These genetically engineered potato plants were used for simultaneous measurements of transient changes in the membrane potential and the Ca(2)(+) luminescence triggered by heat-induced action potentials. High temporal resolution for recording of the fast transient electrical and light signals was accomplished by a sampling rate of 1 kHz. Upon elicitation by heat the membrane potential depolarization preceded the rise of cytosolic Ca(2)(+) by 50-100 ms. Several Ca(2)(+) channel blockers were tested to inhibit the rise in cytosolic Ca(2)(+). Treatment of plants with Ruthenium Red blocked the elevation in cytosolic Ca(2)(+) that was associated with heat-stimulated action potentials. Furthermore, action potentials have been demonstrated to stimulate jasmonic acid biosynthesis and PINII gene expression. Therefore, we measured jasmonic acid and PINII gene expression levels subsequent to action potential initiation by a short heating pulse. As expected, jasmonic acid biosynthesis and PINII gene expression were induced by action potentials. Pretreatment of potato plants with Ruthenium Red inhibited induction of jasmonic acid biosynthesis and PINII gene expression that was generally triggered by heat-activated action potentials. PMID- 15111721 TI - Low phosphorus tolerance mechanisms: phosphorus recycling and photosynthate partitioning in the tropical forage grass, Brachiaria hybrid cultivar Mulato compared with rice. AB - The Brachiaria hybrid cv. Mulato is well adapted to low-fertility acid soils deficient in phosphorus (P). To study the grassy forage's mechanisms for tolerating low P supply, we compared it with rice (Oryza sativa L. cv. Kitaake). We tested by using nutrient solution cultures, and quantified the effects of P deficiency on the enzymatic activities of phosphohydrolases and on carbon metabolism in P-deficient leaves. While P deficiency markedly induced activity of phosphohydrolases in both crops, the ratio of inorganic phosphorus to total P in leaves was greater in Brachiaria hybrid. Phosphorus deficiency in leaves also markedly influenced the partitioning of carbon in both crops. In the Brachiaria hybrid, compared with rice, the smaller proportion of (14)C partitioned into sugars and the larger proportion into amino acids and organic acids in leaves coincided with decreased levels of sucrose and starch. Hence, in P-deficient leaves of the Brachiaria hybrid, triose-P was metabolized into amino acids or organic acids. Results thus indicate that the Brachiaria hybrid, compared with rice, tolerates low P supply to leaves by enhancing sugar catabolism and by inducing the activity of several phosphohydrolases. This apparently causes rapid P turnover and enables the Brachiaria hybrid to use P more efficiently. PMID- 15111722 TI - Comparative analysis of expressed sequence tags in resistant and susceptible ecotypes of Arabidopsis thaliana infected with cucumber mosaic virus. AB - Arabidopsis thaliana ecotype Columbia (Col-0) is susceptible to the yellow strain of cucumber mosaic virus [CMV(Y)], whereas ecotype C24 is resistant to CMV(Y). Comprehensive analyses of approximately 9,000 expressed sequence tags in ecotypes Col-0 and C24 infected with CMV(Y) suggested that the gene expression patterns in the two ecotypes differed. At 6, 12, 24 and 48 h after CMV(Y) inoculation, the expression of 6, 30, 85 and 788 genes, respectively, had changed in C24, as opposed to 20, 80, 53 and 150 genes in CMV(Y)-infected Col-0. At 12, 24 and 48 h after CMV(Y) inoculation, the abundance of 3, 10 and 55 mRNAs was altered in both ecotypes. However, at 6 h after CMV(Y) inoculation, no genes were co-induced or co-suppressed in both ecotypes. This differential pattern of gene expression between the two ecotypes at an early stage of CMV(Y) infection indicated that the cellular response for resistance may differ from that resulting in susceptibility at the level detectable by the macroarray. According to the expression pattern at various stages of infection, the expression of many genes could be grouped into clusters using cluster analysis. About 100 genes that encode proteins involved in chloroplast function were categorized into clusters 1 and 4, which had a differentially lower expression in CMV(Y)-inoculated C24. The expression of various genes encoding proteins in the endomembrane system belonged to clusters 2 and 4, which were induced in CMV(Y)-inoculated C24 and Col-0 leaves. Characterization of CMV(Y)-altered gene expression in the two ecotypes will contribute to a better understanding of the molecular basis of compatible and incompatible interactions between virus and host plants. PMID- 15111723 TI - The role of aquaporin RWC3 in drought avoidance in rice. AB - Although the discovery of aquaporins in plants has resulted in a paradigm shift in the understanding of plant water relations, the relationship between aquaporins and drought resistance still remains elusive. From an agronomic viewpoint, upland rice is traditionally considered as showing drought avoidance. In the investigation of different morphological and physiological responses of upland rice (Oryza sativa L. spp indica cv. Zhonghan 3) and lowland rice (O. sativa L. spp japonica cv. Xiushui 63) to water deficit, we observed young leaf rolling and the remarkable decline of cumulative transpiration in the upland rice. The expression of water channel protein RWC3 mRNA was increased in upland rice at the early response (up to 4 h) to the 20% polyethylene glycol (PEG) 6000 treatment, whereas there was no significant expression changes in lowland rice. Protein levels were increased in upland rice and decreased in lowland rice at 10 h after the water deficit. The up-regulation of RWC3 in upland rice fits well with the knowledge that upland rice adopts the mechanism of drought avoidance. The physiological significance of this RWC3 up-regulation was then explored with the over-expression of RWC3 in transgenic lowland rice (O. sativa L. spp japonica cv. Zhonghua 11) controlled by a stress-inducible SWPA2 promoter. Compared to the wild-type plant, the transgenic lowland rice exhibited higher root osmotic hydraulic conductivity (Lp), leaf water potential and relative cumulative transpiration at the end of 10 h PEG treatment. These results indicated that RWC3 probably played a role in drought avoidance in rice. PMID- 15111724 TI - Simple RNAi vectors for stable and transient suppression of gene function in rice. AB - Since the recent sequencing of the rice genome, the functional identification of rice genes has become increasingly important. Various tagged lines have been generated; however, the number of tagged genes available is not sufficient for extensive study of gene function. To help identify the functions of genes in rice, we developed a Gateway vector, pANDA, for RNA interference of rice genes. This vector can be used for Agrobacterium transformation of rice and allows easy and fast construction of efficient RNAi vectors. In the construct, hairpin RNA derived from a given gene is transcribed from a strong maize ubiquitin promoter, and an intron is placed 5' upstream of inverted repeats to enhance RNA expression. Analysis of rice genes using this vector showed that suppression of mRNA expression was observed in more than 90% of transgenic plants examined, and short interfering RNA indicative of RNA silencing was detected in each silenced plant. A similar vector, pANDA-mini, was also developed for direct transfer into leaf cells or protoplasts. This vector can be used for transient suppression of gene function in rice. These vectors should help identify the functions of rice genes whose tagged mutants are not available at present and complement existing methods for functional genomics of rice. PMID- 15111725 TI - Functional analysis of a rice putative voltage-dependent Ca2+ channel, OsTPC1, expressed in yeast cells lacking its homologous gene CCH1. AB - We isolated a cDNA (OsTPC1) from rice that was homologous to AtTPC1, a putative voltage-dependent Ca(2+) channel (VDCC) gene of Arabidopsis thaliana. The hydropathy profile of its deduced amino acid sequence showed significant structural features of the alpha 1-subunit of animal VDCCs. Functional analysis using a heterologous yeast expression system showed that OsTPC1 facilitates Ca(2+) permeation. The K(m) value for Ca(2+) of OsTPC1, 47.5 micro M, was comparable to that of intrinsic CCH1, a candidate VDCC in yeast. Ca(2+) permeation by OsTPC1 was inhibited by verapamil, a VDCC blocker. These findings indicate for the first time that OsTPC1 is a putative VDCC in rice. PMID- 15111726 TI - Developments in basic science research. Highlights of the 11th Conference on Retroviruses and Opportunistic Infections, February 8-11, 2004, San Francisco, California, USA. AB - As with last year's meeting, perhaps the greatest emphasis in the basic science categories was in the area of cellular cofactors that influence HIV-1 replication either positively or negatively. A number of presentations provided detailed insight into the mechanism by which APOBEC 3G, the cellular target of Vif, effects its antiviral activity. The surprising thing is that this antiviral activity is not restricted to primate lentiviruses but is active against retroviruses and even hepatitis viruses. In the area of positive-acting cellular cofactors, the emphasis was on those cellular proteins that facilitate egress of the virus from the infected cell. It is now apparent that viruses such as HIV-1 can bud into cytoplasmic vesicles in order to establish a unique intracellular reservoir. How viruses move between cells was also the focus of several presentations at the meeting, and there were further surprises about the mechanism by which HIV-1 may establish latent infection. PMID- 15111727 TI - Advances in antiretroviral therapy. Highlights of the 11th Conference on Retroviruses and Opportunistic Infections, February 8-11, 2004, San Francisco, California, USA. AB - Antiretroviral therapy was focus of many of the studies reported at the 11th CROI. This year, data on new drugs, refinements in the management of treatment naive and treatment-experienced persons, the impact of drug resistance (particularly following exposure to a single dose of nevirapine), and the growing experience with antiretrovirals in the developing world were the dominant themes. This reviews summarizes new information relevant to clinicians and clinical researchers. PMID- 15111728 TI - Complications of HIV disease and antiretroviral therapy. Highlights of the 11th Conference on Retroviruses and Opportunistic Infections, February 8-11, 2004, San Francisco, California, USA. AB - This year's conference provided newer insights on the complications of antiretroviral therapy, as well as into the complications that arise from HIV infection itself. Many presentations at the conference centered around metabolic complications of therapy, including lipid abnormalities, diabetes, body composition changes, bone disorders, and cardiovascular disease. New data on complications of HIV infection itself were presented, including those on coinfections with hepatitis B, C, and herpes simplex viruses, malaria, and tuberculosis, as well as complications that are important during pregnancy. This article summarizes these presentations. PMID- 15111729 TI - Modeling women's health with nonhuman primates and other animals. PMID- 15111730 TI - Ovarian dysfunction, stress, and disease: a primate continuum. AB - Menopause is recognized as a period of increased risk for coronary heart disease (CHD) and osteoporosis. Vulnerability to these conditions is often attributed to the naturally occurring estrogen deficiency characteristic of this part of the life cycle. Premenopausal reductions in endogenous estrogen occasioned by functional ovarian abnormalities or failure are hypothesized to be similarly pathogenic and to accelerate development of CHD and osteoporosis prematurely, thereby increasing the health burden of older women. These functional abnormalities, which occur along a continuum from mild, luteal phase progesterone deficiency to amenorrhea, are relatively common and are often attributed to psychogenic factors (stress, anxiety, depression, or other emotional disturbance), exercise, or energy imbalance. Although numerous investigators have commented on these functional deficits, the abnormalities can be difficult to diagnose and are generally unappreciated for the contribution they may make to postmenopausal disease. Studies in nonhuman primates confirm that these deficits are easily induced by psychological stress and exercise, and that they accelerate the development of cardiovascular disease and perhaps bone loss in the presence of a typical North American diet. However, functional reproductive deficits are also reversible and are thus potentially amenable to environmental or behavioral intervention. Data from both women and nonhuman primates support the hypothesis that functional reproductive deficits are adaptive when triggered appropriately but are detrimental when activated in an environment (e.g., sedentary lifestyle, high-fat diet) permissive to the development of chronic disease. PMID- 15111732 TI - Animal studies in endometriosis: a review. AB - Endometriosis is a common women's health problem that is characterized by the presence of tissue resembling endometrium outside the uterus. The condition causes painful periods, chronic pelvic pain, and subfertility, which are potentially debilitating; and it affects millions of women worldwide. The diagnosis is made on visual inspection of the pelvis, usually at laparoscopy. The natural history is unknown, and well-controlled experiments are difficult to perform because of the need for repeated surgical procedures to assess endometriotic lesions over time. Thus, despite over 50 years' research, the cause of endometriosis remains unclear, and treatment options are limited. Animal models provide an invaluable tool to study risk factors, prevalence, and the natural history of endometriosis especially in those menstruating nonhuman primates that develop the disease spontaneously. Many of the practical problems associated with studying the disease in humans can therefore be overcome. The pathophysiology of endometriosis can also be investigated and new treatments assessed in both nonprimates and nonhuman primates, with "disease" induced by placing autologous uterine tissue in ectopic sites, or human endometrium in the case of nude mice. However, although nonprimates have obvious advantages as a model, the extent to which the induced lesions are truly representative of the disease itself is debatable. This review explores the value of the experimental models that have been used to date. PMID- 15111731 TI - Nonhuman primates contribute unique understanding to anovulatory infertility in women. AB - Anovulatory infertility affects a large proportion of reproductive-aged women. Major improvements in successful clinical treatment of this prevalent disorder in women's health have been made possible because of biomedical research employing nonhuman primates. Experiments on female rhesus monkeys were the first to demonstrate that the key hypothalamic neurotransmitter, gonadotropin-releasing hormone, involved in stimulating pituitary gonadotropin synthesis, storage, and release was bioactive only when released in approximately hourly bursts. This breakthrough in understanding gonadotropin regulation enabled identification of hypogonadotropic, apparently normogonadotropic, and hypergonadotropic forms of anovulatory infertility, and development of appropriate stimulatory or inhibitory gonadotropin therapies. Treatments to overcome anovulatory infertility represent one of the major advances in clinical reproductive endocrinology during the last 25 yr. The future promise of nonhuman primate models for human ovulatory dysfunction, however, may be based on an increased understanding of molecular and physiological mechanisms responsible for fetal programming of adult metabolic and reproductive defects and for obesity-related, hyperinsulinemic impairment of oocyte development. PMID- 15111733 TI - Hormone replacement therapy and cardiovascular disease: lessons from a monkey model of postmenopausal women. AB - Concerns exist about the cardiovascular effects of hormone replacement therapy (HRT) in postmenopausal women because results from the Women's Health Initiative (WHI) and the Heart and Estrogen/Progestin Replacement Study (HERS) are contradictory. In both of these studies, postmenopausal conjugated equine estrogens + medroxyprogesterone acetate did not reduce risk, and somewhat increased the risk of myocardial infarction in both primary (WHI) and secondary (HERS) prevention. These results appear to contradict numerous observational clinical trials and animal studies, which reported profound beneficial effects of HRT on cardiovascular disease risk. Results of both human and monkey studies indicate that estrogen replacement therapy (ERT)/HRT is effective in inhibiting progression of early stage (fatty streak) atherosclerosis but that ERT/HRT is much less effective in inhibiting progression of more advanced (established plaque) atherosclerosis. Results of these monkey studies are consistent with those of studies in women wherein ERT/HRT was initiated in postmenopausal women with different initial amounts of atherosclerosis. Based on these findings, it is speculated that ERT/HRT may be more cardioprotective in younger postmenopausal women with less coronary artery disease, and less effective in women with established coronary artery disease. Researchers are challenged to define the relative cardiovascular risk/benefit in different populations of postmenopausal women based on differences in age, amounts of pre-existing atherosclerosis, and risk factors. PMID- 15111734 TI - Stroke in the female: role of biological sex and estrogen. AB - Women are protected from stroke relative to men until the years of menopause. Because stroke is the leading cause of serious, long-term disability in the United States, modeling sex-specific mechanisms and outcomes in animals is vital to research. Important research questions are focused on the effects of hormone replacement therapy, age, reproductive status, and identification of sex-specific risk factors. Available research relevant to stroke in the female has almost exclusively utilized rodent models. Gender-linked stroke outcomes are more detectable in experimental studies than in clinical trials and observational studies. Various estrogens have been extensively studied as neuroprotective agents in women, animals, and a variety of in vitro models of neural injury and degeneration. Most data in animal and cell models are based on 17 beta estradiol and suggest that this steroid is neuroprotective in injury from ischemia/reperfusion. However, current evidence for the clinical benefits of hormone replacement therapy is unclear. Future research in this area will need to expand into stroke models utilizing higher order, gyrencephalic animals such as nonhuman primates if we are to improve extrapolation to the human scenario and to direct and enhance the design of ongoing and future clinical studies and trials. PMID- 15111735 TI - Sex hormones, insulin sensitivity, and diabetes mellitus. AB - Sex differences and the role of gonadal hormones in modulating insulin sensitivity and glucose tolerance are of increasing interest and importance because of the increasing prevalence of type 2 diabetes mellitus and the metabolic abnormalities associated with aging. Body composition is closely associated with insulin sensitivity, and increased body fat, particularly in the visceral compartment, is a risk factor for developing type 2 diabetes mellitus. Sex differences in body composition and/or insulin sensitivity are evident in humans throughout the lifespan. Ovarian hormones influence insulin sensitivity across the menstrual cycle, during pregnancy, and in the menopausal transition. Similarly, estrogens and progestins used for contraception and hormone replacement therapy affect glucoregulation. Nonhuman primates and humans have similar life histories and reproductive characteristics. As a result, nonhuman primates provide a valuable model for investigating factors related to insulin sensitivity. Studies of nonhuman primates have contributed significantly to our understanding of sex differences and the influence of sex steroids in this context. This brief review surveys present knowledge of the sex differences in body composition, insulin sensitivity, and risk for development of type 2 diabetes mellitus derived from studies in humans and nonhuman primates. The influences of endogenous and exogenous gonadal steroids are emphasized. PMID- 15111736 TI - Hormonal therapies and osteoporosis. AB - Osteoporosis, now defined as a disease characterized by low bone mass and a microarchitectural deterioration of bone tissue leading to enhanced bone fragility and fracture risk, is a major public health problem. Classic hormonal therapies to prevent and treat osteoporosis associated with menopause have recently been questioned due to the risk/benefit ratio of prolonged treatment. There is a critical need for safe and effective alternative therapeutics for this disease. Nonhuman primates have been used as models to assess bone changes associated with estrogen deficiency because their trabecular and cortical bone remodeling processes, monthly menstrual cycles, and reproductive-hormone patterns are similar to those of humans. The ovariectomized nonhuman primate has become the preferred model in which to study effects on bone remodeling, particularly with regard to bone mass, architecture, and strength, in fulfillment of studies required by international guidelines for the development of antiosteoporotic drugs. The nonhuman primate is amenable to several methodologies that assess bone quantity and quality, including dual energy x-ray absorptiometry (DXA), quantitative computed tomography (QCT), histology, static and dynamic histomorphometry, and biomechanical testing, as well as assays developed for clinical use, which serve as biomarkers of bone metabolic processes. The use of the nonhuman primate model in the assessment of osteoporosis therapeutics, both hormonal (sex steroids and their analogues, parathyroid hormone) and nonhormonal (bisphosphonates), has provided valuable information on the safety and efficacy as well as the mechanisms of bone loss associated with estrogen deficiency that is directly applicable to the human situation. PMID- 15111737 TI - Neoplasms of the reproductive tract: the role of hormone exposure. AB - Cancers of the reproductive system are a major source of morbidity and mortality among women worldwide. Because the uterus, ovaries, and cervix are hormonally responsive tissues, exposure to endogenous or exogenous sex steroids can profoundly affect the carcinogenic process. Animal models developed to date provide valuable but imperfect systems in which to study neoplasms of the reproductive tract. Nonhuman primate models share the unique primate-specific endometrial physiology of humans, but rarely develop neoplasms of the reproductive tract. Therefore a surrogate marker approach is required for the study of hormonally induced cancer risk in primates. Rodents provide practical models in which tumorigenesis can be assayed in a short time and, with appropriate interpretation, can be used for assessment of risk, prevention, and therapeutic strategies. In addition to the spontaneous strain-dependent incidence of female reproductive cancers, the classical chemical and hormonal carcinogenesis models, and the use of xenograft approaches, novel genetically modified animals provide unique insights into relevant molecular mechanisms. Caveats in the use of rodent models include anatomical differences from the human reproductive tract, the greater possibility of different metabolic responses to hormonal agents than humans, strain variations in tumor type and hormonal responsiveness, and unexpected tumor phenotypes in genetically modified animals. Reported nonmammalian models are limited primarily to the study of ovarian carcinogenesis. Recent progress in the understanding of cervical carcinogenesis is encouraging. Unmet needs in this area of research include models of early events in ovarian carcinogenesis and strongly predictive models of endometrial cancer risk. Nonhuman primates remain indispensable for the study of some aspects of reproductive pathophysiology, but the best understanding of carcinogenesis in the reproductive tract requires a broad approach using complementary human, nonhuman primate, and nonprimate studies. PMID- 15111738 TI - Cognition, mood disorders, and sex hormones. AB - Macaques (Macaca spp.) are useful models to evaluate effects of ovarian sex steroids and selective estrogen receptor modulators (SERMs) on mood and cognitive function due to similarities to women in their reproductive and central nervous systems. The results of nonhuman primate studies support the hypothesis that estrogen mediates specific aspects of attention and memory, yet much work is needed to understand which cognitive processes are affected, whether natural versus surgical menopause effects are different, and the interaction of age and ovarian senescence on cognitive function. This knowledge is necessary to determine whether to support the cognitive function of women in the menopausal phase of life and, if so, to determine efficacious therapeutic interventions. Mood disorders are prevalent in women and are associated with reproductive function in women and macaques. Exogenous steroid therapies, including oral contraceptives and postmenopausal hormone replacement therapies, have behavioral effects in women and appear to affect the behavior and underlying neural substrates of monkeys. Additional research is necessary to confirm and extend these observations. Ovarian steroids have multiple effects on serotonin synthesis, reuptake, and degradation, on neural activity that drives serotonin release, and on receptor activation in primates. This system modulates cognitive function and mood and is the target of a broad class of antidepressant therapies. Understanding the effects of ovarian steroids on the neural serotonergic system is necessary to understand depression in women. These studies are best carried out in primate models, which are more similar to humans in neural serotonergic function than other animal models. PMID- 15111739 TI - Usefulness of the monkey model to investigate the role soy in postmenopausal women's health. AB - Some of the important health issues for postmenopausal women include cardiovascular disease, osteoporosis, breast cancer, and relief of menopausal symptoms. Ovariectomized cynomolgus monkeys (Macaca fascicularis) have many strengths as models for research in this area including a close phylogenetic relationship to humans, similarities in lipid/lipoprotein metabolism and coronary artery anatomy, similar skeletal anatomical and morphological characteristics, mammary glands with similar pathophysiological characteristics, and a 28-day menstrual cycle with similar hormonal fluctuations. Monkeys (macaques) also experience declining ovarian function and irregular menstrual cycles (natural menopause) when they approach 24 to 29 yr of age. However, because of their very short life span after natural menopause, ovariectomized macaques are used to model postmenopausal women. The cynomolgus monkey model has been useful in defining the potential cardiovascular benefits of soy foods and soy supplements; however, it remains unclear whether the observations are generalizable to all women or only to those who, like cynomolgus monkeys, convert the soy isoflavone daidzein to the metabolite equol. Particularly important has been the use of the cynomolgus monkey model to understand the effects of soy on breast health. There is evidence from a cynomolgus monkey trial to suggest that soy/soy phytoestrogens have no estrogen agonist effects for breast. Finally, soy/soy phytoestrogens do not appear to be an adequate alternative to postmenopausal hormone therapy. Nevertheless, important attributes of soy have been identified, and it may have potential as a complementary component to hormone therapy. PMID- 15111740 TI - Role of the nonhuman primate for research related to women's health. AB - This overview of the current status of medical problems that affect women is related to current studies on pathophysiology and therapeutic interventions using nonhuman primates to demonstrate the utility of the primate model for the study of disease processes in women. The current medical literature on women's health is compared with the literature on nonhuman primate research. The findings reviewed in the articles of ILAR Journal Volume 45 Issue 2 of 2004 are evaluated in the context of the scope and problems associated with disease entities in women. Nonhuman primate research with known information regarding women's disease is discussed, and the utility of the animal model for the study of human disease is highlighted, based on its significant relevance due to similarities of nonhuman primate and human subjects' physiology, metabolism, and responses to therapeutic interventions. Additional advantages of the animal model include the ability to control the experimental environment and the capacity to perform chronic study procedures. These findings allow us to utilize the nonhuman primate as the most relevant model in the animal world for the study of human disease processes. PMID- 15111741 TI - Systematic review on the incidence and prevalence of severe maternal morbidity. AB - OBJECTIVE: To summarize the prevalence and the incidence of serious morbidity from studies reporting data on severe maternal morbidity and to compare study designs and definitions. METHODS: A literature search was used to identify relevant studies, which report data on prevalence/incidence of severe complications during pregnancy, delivery and postpartum. For assessment of the quality of studies a structured data collection form from World Health Organization for systematic review of maternal mortality and morbidity was used. Incidence/prevalence and case-fatality ratios were extracted. RESULTS: In this review 24 studies were included, most of them--cross-sectional hospital based (16/24). In ten studies data about one severe maternal condition (admissions to intensive care unit, and hysterectomy) was presented, while fourteen studies dealt with multiple causes of severe maternal morbidity (rupture of uterus, hemorrhage, sepsis, and hypertensive disorders of pregnancy). In these studies very different inclusion criteria due to structure of diseases and severity were used. CONCLUSIONS: The prevalence of severe maternal morbidity ranged from 0.07 8.23% and the case-fatality ratio from 0.02-37%. Studies estimating the incidence of severe maternal morbidity have used different definitions and ways of identification. Severe hemorrhage, sepsis and hypertensive disorders of pregnancy are the commonly used "near-miss" conditions. Further work will be able to create clear definition and method of identification. PMID- 15111742 TI - [Evaluation of combined menisci repair results]. AB - Between 2001 and 2003 sixteen patients underwent repair of the menisci with combined "outside-inside", "all-inside" and reabsorbable arrow techniques. This prospective study compared two patient groups: combined fixation group (16 patients) and "outside-inside" fixation group (16 patients). Patients were evaluated using Lysholm and Tegner activity scales. Results of 12 (87.5%) patients in combined fixation group were excellent and good at the time of last follow-up. Results of 10 (62.5%) patients in "outside-inside" fixation group were excellent and good 10 months post operations. There was statistically significant difference between the groups (p<0.05). PMID- 15111743 TI - [Clinical results after articular cartilage injury revision surgeries]. AB - Twenty-four patients underwent revision surgeries with osteochondral autotransplantation after failed primary articular cartilage restoration procedures in the Clinic of Orthopedics and Traumatology of Kaunas University of Medicine Hospital between 2000 and 2003. Primary microfracture procedure was performed for 22 patients and osteochondral autotransplantation for 2 patients. Patients were evaluated using International cartilage repair society (ICRS) and modified Hospital for Special Surgery (HSS) scales as well as with X-rays. Results of 22 (91.6%) revision osteochondral autotransplantations were excellent and good at the time of last follow-up; results of 2 (8.3%) operations were fair. Modified ICRS and HSS evaluation showed statistically significant improvement of clinical results following 13.2 months after revision osteochondral autotransplantation (p<0.005; p<0.001). PMID- 15111744 TI - [First clinical experience with nonreamed intramedullary nailing in Lithuania]. AB - Long bone diaphyseal fractures are very common. Limited choice of implants for treatment of these fractures in Lithuania often leads to unsatisfactory treatment results and complications. Unreamed intramedullary nailing is relatively simple and effective method for treatment of severely injured patients. Using this method early stabilization and rehabilitation, decreased number of complications, and shortened stay in hospital can be ensured. Indications, technique and early results of first fifty intramedullary fixations with unreamed nails performed in the Clinic of Orthopedics and Traumatology, Kaunas University of Medicine are presented. PMID- 15111745 TI - [Early results of total ankle arthroplasty]. AB - Objective of the study was to evaluate total ankle arthroplasty as the treatment of choice for ankle arthritis; to analyze variables' influence upon early results after total ankle arthroplasty; and to determine early and late postoperative complications and the ways to avoid them. MATERIAL AND METHODS: We evaluated clinical results of 18 patients (out of 23 operated patients), for whom total ankle replacement was performed in 1998-2003. Uncemented Scandinavian Total Ankle Replacement (WLink, Germany) endoprosthesis was used in all cases. The patients were examined, questioned and evaluated according to modified rating system recommended by American Orthopedic Foot and Ankle Society. We observed early and late postoperative complications. RESULTS: Excellent and good results were observed in 9 cases (50%); fair results were in 6 cases (33%), poor in 2 cases (12%), and failure in 1 case (5%). Complications occurred in 11 cases (61%): 4 patients had neurological complaints in operated foot, delayed wound healing was observed in 2 cases, 3 patients had plantar flexion contracture, for 1 patient arthrodesis was done because of dislocation of meniscus component. CONCLUSIONS: Total ankle replacement is an alternative treatment to arthrodesis after posttraumatic ankle arthritis. Results after total ankle arthroplasty are worse than after total hip or knee replacement. Operative technique has great influence on total ankle replacement results. Accurately selected indications for surgery decrease complication rate. PMID- 15111746 TI - [Hip revision arthroplasty (long-term results)]. AB - OBJECTIVE: To evaluate the risk factors after total hip replacement arthroplasty for rerevision and to analyze complications after hip revision surgery. MATERIAL AND METHODS: We obtained data from 117 hip revisions and 12 hip rerevision arthroplasties performed in 1992-2001 in the Department of Orthopedics of Klaipeda Hospital. Special forms were filled in for every patient who participated in the study. Name, operation date, type of implants, operative technique, revision diagnosis, intraoperative and postoperative complications were recorded. All patients were checked for death until 2003. RESULTS: Hip revisions were performed for 77 (66%) women and 50 (44%) men in 1992-2001. We revised 22 (19%) cups, 6 (5%) stems, 86 (74%) total hip revisions; femoral head was exchanged for 3 patients. Revision diagnoses were: aseptic loosening in 106 (90%) cases, recurrent dislocations in 7 (6%) cases, and periprosthetic fractures in 4 (4%) cases. Patients' age varied from 26-82 years, average 63.5 years. In revision group only 8% of patients were less than 50 years old, compared to 33% in rerevision group. Morselized allografts and bone impaction technique for reconstruction of bone defects were used in 70 (60%) of cases. We rerevised one cup only for which revision morselized allografts were used. Eight (67%) rerevisions were performed after first 28 (24%) hip revisions. CONCLUSIONS: Patients, who underwent revision surgery being younger than 50 years old, were at higher risk for rerevision surgery. Revision with morselized bone allografts and bone impaction technique decreases number of rerevisions. Learning curve was steep and had great influence to our results. PMID- 15111747 TI - [Features of the spinal cord injury in distractive flexion and compressive extension cervical spine trauma]. AB - OBJECTIVES: 1) to assess neurological status in patients with distractive flexion and compressive extension cervical spine injuries; 2) to determine the relationship between neurological recovery and the patterns of cervical spine injuries. MATERIAL AND METHODS: Prospectively collected data on 78 persons with traumatic distractive flexion and compressive extension cervical spine injuries. These patients were treated in Kaunas University of Medicine Hospital between 1998 and 2000. The study included 18 (23.1%) females and 60 (76.9%) males. The age range was 16-80 years, mean age was 46.6 years. Cervical spine injuries in all patients were visualized at using Computed tomography and radiography. The cervical spine injury patterns were recognized by the Ferguson-Allen's classification. We divided these patients into two groups: the first group included patients who had sustained distractive flexion cervical spine injuries; the second group included patients who had sustained compressive extension cervical spine trauma. We assessed neurological status of these patients after admission to hospital in the average of four hours after an accident. Motor and sensory evaluation was conducted using the guidelines established by the American Spinal Cord Injury Association. Forty-nine patients were investigated in the average of four years after trauma and treatment. We assessed their neurological status and determined the relationship between neurological recovery and the patterns of cervical spine injury. RESULTS: Forty-eight (61.5%) patients had distractive flexion cervical spine injuries (the first group) and 30 (38.5%) had compressive extension trauma (the second group). We compared patients from the first and the second groups and did not find any difference among an accident circumstances (p>0.05), the level of the cervical spine injury (p>0.05) and neurological status at the early period after a trauma (p>0.05). Eight (16.7%) patients died from the first group, 5 (16.7%) from the second group due to severe spinal cord injury (p>0.05). Thirty patients from the first group, and 19 patients from the second group were investigated in the late posttraumatic period (average four years). We performed statistical analysis of these patients and did not find any difference between the accident circumstance (p>0.05), the level of the cervical spine injury (p>0.05), displacement of the vertebra body (p>0.05), neurological status and did not determine the relationship between neurological recovery and the patterns of cervical spine injuries. CONCLUSIONS: Neurological status of the patients with distractive flexion and compressive extension cervical spine injuries did not differ. The relationship between neurological recovery and the patterns of cervical spine injuries did not exist. PMID- 15111748 TI - [Cervical spine trauma and spinal cord injury: the deaths caused by spinal cord swelling]. AB - OBJECTIVE: To investigate clinical status features of the patient's who died from spinal cord and brain swelling. MATERIAL AND METHODS: Retrospectively collected data on 67 persons with severe spinal cord injury. These patients were treated and died in Kaunas University of Medicine Hospital between 1995 and 2003. The death reasons were determined after autopsy. The study included 2 (3%) females and 65 (97%) males. The age range was 16-82 years, mean age was 49.6 years. Cervical spine injuries in all patients were visualized at computed tomography and radiography. The cervical spine injury patterns were determined using Ferguson-Allen's classification. Motor and sensory evaluation after admission to hospital was conducted using the guidelines established by the American Spinal Cord Injury Association. From this group of patients, 48 cases were selected with spinal cord swelling as the reason of death. We divided these patients into two groups; 21 patients with C4-C5 neurological level were in the first group and 27 persons with C6-Th1 neurological level were in the second group. RESULT: . We compared the patients from the first and the second groups and did not find any difference between their age, the patterns of the accidents, the patterns of the cervical spine injuries (p>0.05), clinical status after admission to hospital and six hours before patient's death (p>0.05). Patients, who died from spinal cord swelling, had alterations in cardiopulmonary function before their death. The patients from the first group died on average in 80 hours after the accident; the patients from the second group died on average in 146 hours after the trauma (p<0.05). CONCLUSIONS: Patients, who died from spinal cord swelling, had distinct respiratory failure before their death. Alterations of the clinical status of patients with C4-C5 neurological level were faster. PMID- 15111749 TI - [Bupivacaine for continuous interscalene brachial plexus analgesia after shoulder surgery]. AB - OBJECTIVE: This study evaluates clinical efficacy of continuous interscalene brachial plexus block with bupivacaine 0.15% for postoperative analgesia after shoulder surgery. MATERIAL AND METHODS: Eighty patients, scheduled for elective shoulder surgery, were included in this study. All patients received an interscalene block before surgery. A perineural catheter was then introduced for continuous brachial plexus analgesia. Patients were prospectively randomized in a double-blinded fashion to receive either bupivacaine 0.15% or placebo postoperative infusion through an interscalene catheter. Pain was assessed with a visual analog scale at rest and also in motion. Duration of surgery and surgery type, satisfaction scores, supplemental analgesia, side effects, circulatory and respiratory parameters were recorded. RESULTS: The bupivacaine group showed less pain at rest and in motion than the placebo group (p<0.0001), except 4 and 6 h after the brachial plexus block; a requirement for supplemental analgesia was also lower. Side effects (except weakness in the arm, which was more frequent in bupivacaine group), circulatory and respiratory parameters were comparable in both groups. Satisfaction scores were higher in bupivacaine group. CONCLUSIONS: Continuous interscalene brachial plexus analgesia is a reliable and effective method of providing postoperative pain relief after shoulder surgery and is superior to the systemic analgesia. PMID- 15111750 TI - [Treatment of early and late obstetric brachial plexus palsy]. AB - The aim of this study was to review and to analyze treatment patterns of early and late obstetric brachial plexus palsy. Eighty-one children with early and late obstetric brachial plexus palsy were treated in the Department of Pediatric Orthopedics and in the Postintensive Care Unit within the period 1988-2002. Children were classified into 2 groups according to age: Ist group (67 newborns) was treated conservatively, and IInd group (14 children with late obstetric brachial plexus palsy with deformity) underwent operative treatment. Active hand movements and innervation were evaluated before and after treatment. Thirty newborns had full recovery, 32 newborns had incomplete recovery, and in 5 cases no improvement was seen. Fourteen children with late obstetric brachial plexus palsy underwent the following operations: rotation osteotomy of the humerus was performed in 10 cases, lengthening of biceps and brachialis muscle tendons--in 6 cases, transposition of triceps muscle tendon--in 1 case, transposition of pectoralis major tendon--in 3 cases and flexor carpi transposition--in 1 case. There was an improvement in active hand movements after operative treatment and rehabilitation. According to our experience, in most cases newborns recover spontaneously or after conservative treatment. Secondary reconstructive surgery of late brachial plexus palsy can improve the condition of these patients. PMID- 15111751 TI - [Importance of conservative treatment and physical load restriction to the course of Osgood-Schlatter's disease]. AB - Osgood- Schlatter's disease is one of most common causes of knee pain in young patients. In 2000-2002, 171 young patients were consulted for Osgood-Schlatter's disease in the Departments of Orthopedics and Traumatology in Kaunas University of Medicine. There were 41 (24%) females and 130 (76%) males; the age ranging from 10 to 27 years. Patients were divided into two groups: the first group (92 patients) underwent conservative treatment and physical load restriction; the second group (79 patients) had no physical load restriction or they neglected reduced physical load guidelines. In the first group 52 patients were treated by conservative measures and physical load restriction, while 40 patients had only physical load restriction. Pain disappeared faster (approximately after 13 months) in the first group patients, who were treated conservatively and had reduced physical load, compared to patients, who had only physical load restriction (pain disappeared approximately after 15 months), p<0.05 (p=0.0008). Pain disappeared faster in patients who had only physical load restriction (approximately after 15 months) compared to the second group patients who had no physical load restriction (approximately after 16.5 months), p<0.05 (p=0.006). There was 1 complication (encapsulated ossification producing pain) in the first group, compared with 8 complications in the second one (2 patients with 3 type avulsion fracture, 1 patient with 2 type avulsion fracture, and 5 patients with encapsulated ossification producing pain). Complications were treated intraoperatively. Physical load restriction is important for beneficial course of Osgood-Schlatter's disease and prevention of complications. PMID- 15111752 TI - [Efficiency of new treatment methods in burned patients]. AB - The aim of study was to investigate if new treatment methods reduced mortality and diminished hospital stay time for survivors. MATERIAL AND METHODS: We assessed patients, treated in Kaunas University of Medicine Hospital with 10-80% 2B-3 degrees body surface area burned. Group A, which was assessed prospectively, included 89 patients, treated during 2001-2003. All patients in Group A received sufficient enteral nutrition, in 21 patient early escharectomy and skin-grafting were done. This group was compared with Group B, assessed retrospectively and included 89 patients, treated in 1997-1998. They did not receive enteral nutrition and were treated without early escharectomy and skin grafting. RESULTS: Groups were homogeneous by age, extent of general and deep burn, and burn indexes. For 24% patients in Group A early escharectomy and skin-grafting was done, they were operated on the average at 3.6 day after admission (standard deviation - 1.32). In Group A mortality was 5.6%. Average hospital stay time for survivors was 35.7 days (standard deviation--20.4) or 0.9 day/% body surface area burned, standard deviation--0.6. In Group B no early escharectomies and skin grafting were done. Mortality in Group B was 23.6%, hospital stay time for survivors--40.6 days (standard deviation--23.6), or 1.9 day/% body surface area burned. These indicators were statistically significantly different compared to Group A (p<0.05). The significant influence of new treatment methods in major burns was established. CONCLUSION: New methods, early escharectomy and skin grafting and enteral nutrition, introduced in Kaunas University of Medicine Hospital in the last 5-7 years significantly reduced mortality and hospital stay time for survivors (p<0.05). PMID- 15111753 TI - [Analysis of burn-related deaths in Kaunas University of Medicine Hospital during 1993-2002]. AB - Objective of this study was to investigate mortality of burned patients, treated in Kaunas University of Medicine Hospital during 1993-2002, changes of mortality, causes of death, to assess patients' age, gender, burn agent, and adjacent diseases. MATERIAL AND METHODS: Retrospective analysis of case-records of 283 burned patients deceased during 1993-2002 was done. RESULTS: During 1993-2002, 1876 burned adult patients were hospitalized in Kaunas University of Medicine Hospital. The mortality rate of burned patients was between 9 and 22% (average- 13.3%, standard deviation--3.8). Age of deceased patients was on average 56 years (standard deviation--8); actually 21.6% were older that 80 years. There were 62% men among deceased burned patients. Common body surface area burned was 32% (standard deviation--28.6%), deep burn area was at average 22% (standard deviation--19.8%). Seventy two percent of burns were caused by fire, and 10% of patients were scalded. In 35% case-records adjacent diseases were not mentioned, in 57% atherosclerosis and ischemic heart disease were diagnosed, 5% of patients had respiratory diseases, 7% had central nervous system troubles, mental disorders were diagnosed in 2%. Eight percent were cachectic at admission, 6%- with chronic alcohol dependence. In 70% of patients pneumonia was diagnosed, in 13%--pulmonary edema, and in 39%--sepsis. Deceased patients were treated until death on average 14 days (standard deviation--6); during first two weeks 50% died. CONCLUSIONS: At higher mortality risk are elder burned patient with major burns, especially with serious adjacent diseases. Common death causes in burned patients are pneumonia, pulmonary edema and sepsis. PMID- 15111754 TI - [Primary non-Hodgkin's lymphoma of bone]. AB - Primary lymphoma of bone is a rare condition that has been described as a malignant neoplasm formed of lymphoid and myelopoetic tissues. Morphologic substrate is formed of lymphoid cells of different evolutional stages. Primary lymphoma of bone occurs predominantly in males; a male to female ratio is 1.8:1. It may occur at any age. The sites commonly affected are the long bones. In Lithuania 400 new cases of lymphoma are diagnosed every year. About 75% of them are non-Hodgkin's lymphoma. Many histological types and subtypes of lymphoma exist. This variation leads to difficulties when reviewing the literature and comparing prognoses, treatment and oncologic outcomes. Treatment options for primary bone lymphoma historically have included local therapy with radiation or surgery with or without systemic therapy. The aim of surgery is prophylactic fixation of impending fractures or treatment of pathological fractures. We present a clinical case report of 40-year-old male who developed primary lymphoma of bone with pathological fracture of femur. The systemic chemotherapy was applied during the time of treatment. The choice of surgical treatment was intramedullary nailing without using of methylmethacrylate, because regeneration of bone was visible on roentgenograms. After twelve-month follow-up the patient is full weight bearing without external support. PMID- 15111755 TI - Skin cancer prevention: children's health education on protection from sun exposure and assessment of its efficiency. AB - The aim of the study was to assess schoolchildren's knowledge on sun exposure, the peculiarities of behavior in the sun, and the possibility of altering these indicators via education program. The study of the efficiency of the self designed educational program "Let's know the sun better" included 213 fifth grade pupils (113 boys and 100 girls) from Kaunas city schools; the pupils were differentiated into two groups: the experimental (n=106) and the control (n=107) groups. The method employed was anonymous questionnaire-based inquiry. The data of the inquiry applied before the application of the educational programs showed that schoolchildren's behavior in the sun is careless: 40.4% of the studied schoolchildren experienced severe sunburns; 54.0% of children spend three and more hours on the beaches, most frequently between 11 am and 3 pm. Not all schoolchildren use sun protection measures. Most frequently they only have bathing suits (70.0%), baseball caps (56.8%), and sunglasses (57.7%). Out of the studied schoolchildren 18.8% use sunscreen, but only 7.3% of them know how to use it properly. The second inquiry was performed after the schoolchildren were able to apply the recommendations of the educational program "Let's know the sun better" in practice during the summer. The data of this inquiry showed that the knowledge, attitudes, and the peculiarities of behavior in the sun in the experimental group were better compared to the control group. Significantly more schoolchildren in the experimental group (44.1%), compared to the control group (8.5%), used sunscreens properly (p<0.05) and knew which sunscreen is the most suitable (respectively, 42.3% and 20.6%; p<0.05); in addition to that, the children in the experimental group more frequently wore long-sleeved shirts on the beaches (21.0% and 7.5%, respectively; p<0.05), wide-brimmed sunbonnets (37.1% and 10.4%; p<0.05), and sunglasses (61.9% and 44.3%; p<0.05). The findings of the study proved both the necessity and the efficiency of the prepared educational program. PMID- 15111756 TI - [The guidelines for diagnostics and treatment of cervical cancer]. AB - Cervical cancer is one of the most common cancers in women. The purpose of this article is to analyze the main diagnostic and treatment strategies for all stages and recurrences of cervical cancer. The article reviews the epidemiological situation, clinical features, diagnostic procedures for detection of this tumor and for evaluation of the dissemination of the disease, staging criteria, TNM (Tumor, Nodes, Metastases) and FIGO (Federation Internationale de Gynecologie et d'Obstetrique) classification, as well as treatment and prognosis. Surgical treatment (radical type II or III hysterectomy and lymphonodectomy) for early stage I and IIA cervical cancer is the main treatment method. Delivery of adjuvant postoperative radiation therapy or concomitant chemoradiation depends on the prognostic factors (tumor penetration to cervical tissues, lymphovascular invasion, tumor invasion to paracervical tissues, and surgical margins). For treatment of more advanced stages of cervical cancer (IIB, IIIA, IIIB, IVA) concomitant chemoradiation: external beam radiotherapy with chemotherapy and brachytherapy is used. Description of the treatment guidelines for each stage of cervical cancer is given in this article. These guidelines are useful for good treatment practice. PMID- 15111757 TI - Determinants of participation in treatment decision-making by older breast cancer patients. AB - PURPOSE: To identify the impact of patient age and patient-physician communication on older breast cancer patients' participation in treatment decision-making. METHODS: We conducted a cross-sectional survey of breast cancer patients aged 55 years or older (n = 222) in Los Angeles County. Patients received a breast cancer diagnosis between 1998 and 2000, and were interviewed on average 7.1 months (SD = 2.9) from diagnosis. All patient-physician communication variables were measured by patient self-report. Patient participation in treatment decision-making was defined by (1) questioning the surgeon about treatment, and (2) perception of self as the final decision-maker. RESULTS: In multiple logistic regression analyses, surgeons' specific solicitation of patients' input about treatment preferences had positive relationships with both dimensions of patient participation in decision-making, that is, questioning the surgeon (adjusted odds ratio [OR] = 2.09, 95% confidence interval [CI] = 1.05 4.16) and perceiving oneself to be the final decision-maker (OR = 2.38, CI = 1.08 5.28), controlling for patients' sociodemographic and case-mix characteristics and social support. Greater emotional support from surgeons was negatively associated with patient perception of being the final decision-maker. Physicians' information-giving and patient age were not associated with the participation measures. However, greater patient-perceived self-efficacy in patient-physician interactions was related to participation. CONCLUSION: In breast cancer patients aged 55 years and older, surgeons' solicitation of patients' treatment preferences was a powerful independent predictor of patient participation in treatment decision-making, as was patient's self-efficacy in interacting with physicians. Increasing both physicians' and patients' partnership-building skills might enhance the quality of treatment decision-making and treatment outcomes in this burgeoning patient population. PMID- 15111758 TI - Magnetic resonance imaging in comparison to clinical palpation in assessing the response of breast cancer to epirubicin primary chemotherapy. AB - PURPOSE: To investigate whether magnetic resonance imaging (MRI) is superior to clinical palpation in the assessment of response of breast cancer to primary chemotherapy (PC). PATIENTS AND METHODS: Seventy-three patients with T2-4, N0, M0 breast cancer were treated with 3-4 cycles of single agent epirubicin before definitive surgery. MRI was performed at baseline condition and at the end of chemotherapy. RESULTS: According to the WHO criteria, 20 (27.4%) patients attained a complete response (CR) by clinical palpation and 41 (56.2%) a partial response. The corresponding response rate by MRI was 11 (15.1%) and 34 (46.6%), respectively. Residual tumor assessed by MRI better correlated with pathologic measurements (Spearman r : 0.72) than residual tumor assessed by clinical palpation (Spearman r : 0.58). Post-chemotherapy histology evaluation revealed pathologic CR in three cases, only one of them was considered as complete responder by MRI. Residual disease consisted in in situ carcinoma in four cases, one of them was complete responder at MRI, the remaining three showed residual abnormal contrast enhancement indistinguishable from that of invasive tumors. CONCLUSIONS: As compared to pathology specimens, MRI is able to represent the extent of cancer more accurately than clinical palpation. It constitutes a promising technique in assessing the BC response to PC. The current limit of MRI is the scarce specificity in predicting the nature of residual disease. PMID- 15111759 TI - The full potential of breast cancer screening use to reduce mortality has not yet been realized in the United States. AB - OBJECTIVE: Breast cancer mortality has been declining in European countries and the United States since the early 1990s. Based on breast cancer screening programs in western European countries, the reduction in mortality results from a predictable pattern of increasing early-stage and subsequent declining incidence of late-stage cancers. The purpose of this study was to determine whether changes in the incidence of early-stage and late-stage breast cancers has occurred in the United States to suggest that a reduction in breast cancer mortality is the result of screening. METHOD: The analyses are based on women 50-69 years of age using 1990-1998 Surveillance, Epidemiology, and End Results data. Five indicators that are precursors to reductions in mortality are described: in situ breast cancer, T1 tumors (< 2 cm), stage II-IV tumors, lymph node-positive cancers, and locally advanced breast cancers (LABC). RESULTS: The rate of in situ tumors increased from 37.8 to 67.0 per 100,000 population and that of T1 tumors increased from 143.5 to 163.5 per 100,000 population during 1990-1998. The rates of stage II-IV tumors, lymph node-positive cancers, and LABC remained unchanged at about 120 per 100,000, 76 per 100,000, and 17 per 100,000 population, respectively. CONCLUSIONS: Although there has been an increase in early-stage breast cancers (in situ and T1 tumors), the prerequisite decline in late-stage cancers has not yet occurred in the United States--a pattern that was observed in European studies. Possible explanations include the lack of widespread mammography use during the 1980s and, therefore, insufficient elapsed time since mammography use has become more widespread. PMID- 15111760 TI - The influence of clinical information on the accuracy of diagnostic mammography. AB - OBJECTIVE: This study examined the influence of knowledge of clinical information on the accuracy of mammography in women referred for investigation of breast symptoms. METHODS: Subjects were sampled from all women consecutively attending a symptomatic breast clinic and aged 25-55 years. This included all 240 women shown to have breast cancer and 240 age-matched women shown not to have cancer. Mammography films were prospectively reported by two radiologists independently of each other in a blinded manner and without knowledge of any clinical information. The films were then re-read with information about the type and site of symptoms (and without knowledge of the level of suspicion of cancer on clinical examination). The accuracy of reading with and without information on symptoms was compared using sensitivity and specificity and receiver operating characteristic (ROC) curves. The effect of age on changes in test accuracy was examined. RESULTS: Reporting the mammogram with knowledge of clinical information (compared to without any information) significantly improved sensitivity (75.8 vs. 71.3%, P = 0.003) for one radiologist, with a non-significant reduction in specificity (85.4 vs. 87.1%, P = 0.22). For the other radiologist, it resulted in non-significant improvement in both sensitivity (75.4 vs. 73.8%, P = 0.13) and specificity (89.2 vs. 87.9%, P = 0.25). Age did not have a statistically significant effect on changes in test accuracy in our data. ROC curves for both radiologists showed that reporting mammography with knowledge of clinical information resulted in small (about 2%) but significant improvement in overall test accuracy. CONCLUSION: Our findings support the provision of clinical information relating to patients' presentation to radiologists reporting diagnostic mammography. PMID- 15111761 TI - Decreased secretion of Cathepsin D in breast cancer in vivo by tamoxifen: mediated by the mannose-6-phosphate/IGF-II receptor? AB - The lysosomal protease Cathepsin D (Cath D) is associated with increased invasiveness and metastasis in breast cancer. Both estrogen and tamoxifen have been reported to increase Cath D, which seems to contradict the efficacy of tamoxifen as an adjuvant for estrogen dependent breast cancer. Cath D is bioactive in the extracellular space but very little is known about hormonal regulation of secreted Cath D in vivo. In this study we used microdialysis to sample the extracellular fluid in estrogen receptor positive MCF-7 tumors in nude mice. We show that tamoxifen in combination with estradiol decreased secreted Cath D compared with estradiol treatment only in solid tumors in situ. Cell culture of MCF-7 cells revealed that estradiol and tamoxifen increased intracellular proteolytic activity of Cath D in a similar fashion whereas secretion of Cath D was increased by estradiol and inhibited by tamoxifen. Immunofluorescence showed that estradiol located Cath D to the cell surface, while tamoxifen accumulated Cath D to dense lysosomes in perinuclear regions. Moreover, tamoxifen increased the intracellular transporter of Cath D, the mannose 6-phosphate/IGF-II receptor (M6P/IGF2R). In contrast, estradiol decreased the levels of this receptor. Thus, secretion of Cath D is hormone dependent and may be mediated by altered expression of the M6P/IGF2R. Our results highlight the importance of measurements of proteins in all compartments where they are biological active and show that microdialysis is a viable technique for sampling of Cath D in vivo. PMID- 15111762 TI - Genetic polymorphisms in uridine diphospho-glucuronosyltransferase 1A1 (UGT1A1) and risk of breast cancer. AB - Uridine diphospho-glucuronosyltransferase 1A1 (UGT1A1) is involved in catalyzing estrogen, the hormone that plays a central role in the etiology of breast cancer. A common polymorphism [A(TA)6TAA (allele *1) to A(TA)7TAA change (allele *28)] in the TATA-box of the promoter region of the UGT1A1 gene has been reported to be associated with a reduced transcription of this gene. We investigated the association of this polymorphism with the risk of breast cancer among 1047 breast cancer cases and 1083 community controls in the Shanghai Breast Cancer Study, a population-based case-control study. Approximately same proportion of cases (12.5%) and controls (13.0%) carried the variant allele *28 in the Chinese population (p = 0.32). When stratified by age, carrying the *28 allele was associated with an increased risk of breast cancer among women aged less than 40 years (odds ratio [OR] = 1.7; 95% CI = 1.0-2.7) but not among women 40 years old and over (OR = 0.8; 0.7-1.1). Only a few women were homozygous for the *28 allele, precluding a detailed gene-dose association analysis. Additional analyses showed that, the elevated risk associated with the UGT1A1 *28 allele among young women was primarily seen in women who had a later menarche, short menstrual years, absence of family history of breast cancer, low waist-to-hip ratio, or low body-mass index. These results suggested that the *28 allele in the UGT1A1 gene may be associated with an increased risk for breast cancer among Chinese women under age 40. No significant associations were observed with *28 allele and breast cancer risk by estrogen receptor/progesterone receptor status. PMID- 15111763 TI - Anastrozole ('Arimidex') versus tamoxifen as first-line therapy in postmenopausal women with advanced breast cancer: results of the double-blind cross-over SAKK trial 21/95--a sub-study of the TARGET (Tamoxifen or 'Arimidex' Randomized Group Efficacy and Tolerability) trial. AB - It is desirable to identify the most effective sequence of endocrine therapies for the treatment of postmenopausal women with advanced, hormone-responsive breast cancer. In a retrospective analysis of two large, randomized, comparative Phase III trials in this patient population, the Tamoxifen or 'Arimidex' Randomized Group Efficacy and Tolerability (TARGET) trial and the North American trial, we ascertained that tumors responding to anastrozole as first-line therapy may subsequently respond to tamoxifen as second-line therapy. In a double-blind cross-over trial, the SAKK 21/95 sub-trial (including patients from the Swiss centres in the TARGET trial), we further investigated the clinical impact of anastrozole followed by tamoxifen compared with that of tamoxifen followed by anastrozole. Patients with locally advanced or metastatic breast cancer who had continued on randomized treatment until objective disease progression and were still considered suitable for endocrine therapy, could continue on blinded therapy crossing-over to the alternative treatment. They were assessed for time to progression (TTP) from treatment randomization, TTP after crossing-over treatments, time from randomization to progression after crossing-over treatments and overall survival. Median TTP from randomization for patients receiving first line treatment with anastrozole (n = 31) and tamoxifen (n = 29) was 11.3 and 8.3 months, respectively, p = 0.75. Median TTP from treatment cross-over was 6.7 months for tamoxifen after progression on anastrozole (n = 19) and 5.7 months for anastrozole after progression on tamoxifen (n = 18), while median time from randomization to second progression was 28.2 and 19.5 months, respectively. Overall survival from randomization for the anastrozole-tamoxifen sequence and the tamoxifen-anastrozole sequence was 69.7 versus 59.3 months, respectively, p = 0.10. The relative risk of death was higher for the tamoxifen followed by anastrozole sequence (1.63; 95% confidence interval [CI] 0.89-2.98). Tamoxifen is an effective second-line therapy after anastrozole. Our data, together with the better tolerability profile of anastrozole compared with tamoxifen, support the use of anastrozole as first-line therapy for advanced breast cancer in postmenopausal women with hormone receptor-positive tumors. Treatment with tamoxifen may still be useful upon subsequent progression. PMID- 15111764 TI - Phenotypic similarities in bilateral breast cancer. AB - Bilateral breast cancers that develop at similar times in an individual are likely to have been subjected to similar hormonal, environmental and genetic influences during tumourogenesis compared with metachronous tumours. As such, it is possible that tumour phenotype in synchronous bilateral breast cancer may display similar biological characteristics. The aim of this study was to identify phenotypic similarities between synchronous and metachronous bilateral breast cancers which may suggest a common origin. Thirty-three cases of synchronous and 46 cases of metachronous bilateral breast cancer that displayed similar tumour type were analysed for concordance in relation to various histological and immunohistochemical parameters. A higher level of concordance was demonstrated for synchronous cases with the highest level seen for oestrogen receptor. It is likely that this is related to similar tumourogenic pathways occurring at equivalent exposure times to various environmental and hormonal influences, although, in a proportion of cases, inherited genetic factors may play a role. PMID- 15111765 TI - Use of complementary and alternative medicine by chinese women with breast cancer. AB - The use of complementary and alternative medicine (CAM) has been rapidly increasing among cancer patients. The aim of this study is to evaluate the prevalence and patterns of CAM use, particularly patients' intentions and their perceived effectiveness of using Chinese herbal medicine (CHM), as well as the relations between the herbal medicine use and demographic and clinical factors among Chinese women with breast cancer. We analyzed the data from a population based sample of 1065 breast cancer women in urban Shanghai. Patients' average age at diagnosis was 48.1 years and the median time from the initial diagnosis to the follow-up survey was 4.3 years. Overall, 98% of patients had used at least one form of CAM therapy after diagnosis of breast cancer. The most popular CAM modality was traditional Chinese medicine (86.7%), followed by the use of supplements (84.8%), physical exercises (65.5%), and support group attendance (16.6%). CHM was used by 86.4% of patients, while acupuncture was used only by 4.9% of patients. Treating cancer (81.5%) was the most common intentions of using CHM. Other cited intentions included enhancing the immune system (12%), preventing metastasis of cancer or managing other discomforts (7.9%), and lessening menopausal symptoms (4.7%). The majority of patients reported that they had benefited from the use of CHM. Patients who were younger, married, had higher education or income, received chemotherapy or radiotherapy, or had recurrence/metastasis of cancer tended to use CHM more frequently than other patients. The relations between patient characteristics and use of CHMs varied with users' intentions. Given the high prevalence of CAM use among breast cancer patients, research is urgently needed to systematically evaluate the efficacy and safety of CAM use, particularly use of herbal medicines. PMID- 15111766 TI - A systematic review of intraoperative radiotherapy in early breast cancer. AB - A systematic review was undertaken to assess the safety and efficacy of intraoperative radiotherapy (IORT) in early breast cancer compared with breast conserving surgery with postoperative radiotherapy. Literature databases were searched up to March 2002 inclusive. IORT studies of any design and breast conserving therapy randomised controlled trials with sample sizes greater than 500 patients (with at least one arm of breast conserving therapy (BCT)) and systematic reviews on BCT published since 1992, were included for comparison. Seven unique IORT studies were located; one randomised controlled trial, one study with a concurrent control group and five case series. Minor postoperative complications were reported following IORT. Short-term results were similar for both treatment modalities in terms of local recurrence, disease-free and overall survival. However, the current evidence base is poor, making definitive assessment on IORT very difficult. Further research is required to clarify several issues such as identification of the most appropriate subgroups of patients for IORT, a comparison of the currently available mobile IORT technologies, establishing whether IORT is most appropriate as a boost replacement dose or replacement for all postoperative radiotherapy, the examination of how biological repair processes may differ between the two treatment modalities and determining precisely where local recurrences originate with respect to the original tumour site. PMID- 15111767 TI - Three-dimensional in vitro tissue culture models of breast cancer-- a review. AB - Three-dimensional (3D) in vitro breast tumour models have an invaluable role in tumour biology today providing some very important insights into breast cancer. As well as increasing our understanding of homeostasis, cellular differentiation and tissue organization they provide a well defined environment for cancer research in contrast to the complex host environment of an in vivo model. With the recent availability of relevant stromal elements together with the vast array of extracellular matrix constituents available, in vivo like microenvironments can be recreated. These tissue like structures more realistically model the structural architecture and differentiated function of breast cancer than a cellular monolayer providing in vivo like responses to therapeutic agents. Three dimensional in vitro models allow the study of cell-cell and cell-extracellular matrix interactions, in addition to the influence of the microenvironment on cellular differentiation, proliferation, apoptosis and gene expression. Due to their enormous potential 3D cultures are currently being exploited by many other branches of biomedical science with therapeutically orientated studies becoming the major focus of research. In return great progress in 3D culture techniques have been made, largely due to this greater interaction. At present they are being used in studies ranging from investigating the role of adhesion molecules (e.g., E-cadherin) in invasion/metastasis; VEGF and angiogenesis, to tissue modelling and remodelling. Progress in the development of complex 3D culture systems is more productive than ever, however further research is vital. PMID- 15111768 TI - Naringenin inhibits glucose uptake in MCF-7 breast cancer cells: a mechanism for impaired cellular proliferation. AB - Certain flavonoids inhibit glucose uptake in cultured cells. In this report, we show that the grapefruit flava-none naringenin inhibited insulin-stimulated glucose uptake in proliferating and growth-arrested MCF-7 breast cancer cells. Our findings indicate that naringenin inhibits the activity of phosphoinositide 3 kinase (PI3K), a key regulator of insulin-induced GLUT4 translocation, as shown by impaired phosphorylation of the downstream signaling molecule Akt. Naringenin also inhibited the phosphorylation of p44/p42 mitogen-activated protein kinase (MAPK). Inhibition of the MAPK pathway with PD98059, a MAPK kinase inhibitor, reduced insulin-stimulated glucose uptake by approximately 60%. The MAPK pathway therefore appears to contribute significantly to insulin-stimulated glucose uptake in breast cancer cells. Importantly, decreasing the availability of glucose by lowering the glucose concentration of the culture medium inhibited proliferation, as did treatment with naringenin. Collectively, our findings suggest that naringenin inhibits the proliferation of MCF-7 cells via impaired glucose uptake. Because a physiologically attainable dose of 10 micro M naringenin reduced insulin-stimulated glucose uptake by nearly 25% and also reduced cell proliferation, naringenin may possess therapeutic potential as an anti-proliferative agent. PMID- 15111769 TI - Regulation of the estrogen receptor alpha minimal promoter by Sp1, USF-1 and ERalpha. AB - The exact molecular mechanisms regulating estrogen receptor alpha (ERalpha) expression in breast tumors are unclear, but studies suggest that they are partly at the level of transcription. We have focused on the transcription factors that regulate the ERalpha minimal promoter, which we have previously shown to reside within the first 245 bp of the 5'-flanking region of the gene. Within this region are several elements essential for full ERalpha promoter transcriptional activity, including a GC box and an imperfect E box. In earlier studies we demonstrated an essential function for the Sp1 family of transcription factors in the regulation of ERalpha expression. We have now identified both USF-1 and ERalpha itself as components of a multi-protein complex of transcription factors that interacts at the ERalpha minimal promoter and is essential for its full transcriptional activity. Electrophoretic mobility shift assays demonstrated that Sp1 and USF-1, but not ERalpha, bind directly to the ERalpha minimal promoter. We showed by GST pull-down assays that ERalpha is able to interact in vitro with USF 1, suggesting, in addition to a possible interaction between ERalpha and Sp1, a mechanism whereby ERalpha is able to interact with the protein complex. Combined exogenous expression of the components of the complex in MCF-7 breast cancer cells resulted in a synergistic effect on transactivation of the ERalpha minimal promoter, suggesting that the importance of the protein complex is in the interactions among the components. Based upon these findings, we propose a possible model for transcription from the ERalpha minimal promoter. PMID- 15111770 TI - A family-based genetic association study of variants in estrogen-metabolism genes COMT and CYP1B1 and breast cancer risk. AB - In this paper, we report findings from a family-based association study examining the association between polymorphisms in two key estrogen-metabolism genes CYP1B1 (codon 432 G --> C and codon 453 A --> G variants) and COMT (codon 158 G --> A variant) and female breast cancer. We conducted the study among 280 nuclear families containing one or more daughters with breast cancer with a total of 1124 family members (702 with available constitutional DNA and questionnaire data and 421 without). These nuclear families were selected from breast cancer families participating in the Metropolitan New York Registry (MNYR) - one of the six centers of NCI's Breast Cooperative Family Registry. We used likelihood-based statistical methods to examine the allelic associations. We found none of the variant alleles of the CYP1B1 and COMT genes to be associated with breast cancer in these families. This was consistent with results from matched case-control analyses using all available sib-ships in these families. However, we found that parental carrier status of the CYP1B1 codon 453 variant G allele and the COMT codon 158 variant A allele was associated with breast cancer risk in daughters (independent of the daughters' own genotype). In conclusion, findings from this family-based study indicate that a woman's own CYP1B1 or COMT genotypes are not associated with her breast cancer risk. Although the study found that parental carrier status of certain CYP1B1 or COMT genotypes might be associated with daughter's breast cancer risk, the biological basis as well as independent confirmation of this finding need to be investigated in future larger family based studies before making meaningful inferences. PMID- 15111771 TI - Hormonal factors and breast tumor proliferation: do factors that affect cancer risk also affect tumor growth? AB - BACKGROUND: Tumor cell proliferation is one of the most significant predictors of prognosis for women with breast cancer. Personal characteristics that affect hormonal exposure have been implicated in breast tumor etiology, and it is possible that they may also influence tumor cell proliferation. We examined the association between hormone-related breast cancer risk factors and breast tumor proliferation, as reflected in two proliferation measures, Ki-67 and mitotic count. METHODS: The study population was 484 women 40 years of age and older, who were members of a managed care organization's breast cancer screening program and were diagnosed with invasive breast cancer between 1988 and 1995. The percent of Ki-67 positive tumor cells averaged over four high powered fields (Ki-67) was log transformed and analyzed in a linear regression model. Mitotic count was dichotomized into high versus low ( 0.05). Infants whose mothers had ID or IDA during the study period were more likely to develop ID or IDA independently from iron supplementation. Serum ferritin levels decreased between 4 and 6 months of age in the control and daily groups; the weekly group showed no such decrease. In all groups, the mean levels of serum ferritin were significantly increased from 6 months to 7 months of age during the weaning period. In this study, which had a limited number of cases, weekly or daily iron supplementation was not found to decrease the likelihood of IDA. In conclusion, exclusively breast-fed infants with maternal IDA appeared to be at increased risk of developing IDA. PMID- 15111780 TI - Serum lipid alterations in acute lymphoblastic leukemia of childhood. AB - Epidemiologic studies have indicated a relationship between serum lipids and cancer, and it is possible that lipid abnormalities are involved in the mechanism of oncogenesis. This study was performed to investigate serum lipid alterations in patients with acute lymphoblastic leukemia (ALL) at diagnosis and during remission of the disease. Plasma lipids and lipoproteins were measured at diagnosis, prior to the administration of induction treatment, and every 2 months for the first 12 months of the maintenance phase of chemotherapy in 64 patients with ALL. Nearly all patients demonstrated a predictable pattern of serum lipid alterations that consisted of extremely low levels of high-density lipoprotein cholesterol, elevated triglycerides, and elevated low-density lipoprotein cholesterol. Patients studied again during remission demonstrated a return to normal values, and the difference was statistically significant. The results suggest that at diagnosis of ALL an abnormality in lipid metabolism is present, which is reversed during remission. PMID- 15111781 TI - A strategy to detect chromosomal abnormalities in children with acute lymphoblastic leukemia. AB - Conventional cytogenetics (CC) can be used to identify chromosomal abnormalities that are predictors of treatment outcome in acute lymphoblastic leukemia (ALL). The detection of abnormalities in ALL is difficult because low mitotic index and poor-quality metaphases are obtained. Flow cytometry (FC) and fluorescence in situ hybridization (FISH) can be used to detect aneuploidy in any phase of the cell cycle, increasing the number of analyzable cells. The aim of this study was to develop a strategy combining these methods to improve the frequency of chromosome abnormality detection. One hundred children with newly diagnosed ALL were included. CC and DNA content analysis by FC were performed in all patients. The numerical abnormalities identified by both methods were compared and patients were classified as concordant or discordant. FISH was used to support aneuploidy results in discrepant cases using centromeric probes for the chromosomes most frequently involved in aneuploidy. CC and FC showed high concordance (86%). Fourteen cases were discrepant: nine showed hypodiploidy and low hyperdiploidy by cytogenetics and five showed high hyperdiploidy by FC. FISH confirmed aneuploidy in 12 cases in which it could be performed. High hyperdiploidy was the most common abnormality; the 31 cases showing this aneuploidy were identified by FC. The search for abnormalities must begin by measuring DNA content to detect this aneuploidy, which is useful to evaluate the patient's risk. However, it is important to screen for structural abnormalities by CC or molecular techniques. This strategy may detect chromosomal abnormalities, optimizing resources in laboratories where not all the screening methods are available. PMID- 15111782 TI - Suppression of Hodgkin's disease in a patient with Cushing's syndrome. AB - Hodgkin's disease, one of the more common tumors in pediatric oncology, has been shown to be highly responsive to steroids in conjunction with other chemotherapy agents. The authors describe a patient with Cushing's syndrome that suppressed his Hodgkin's disease. He developed constitutional symptoms, and his Hodgkin's disease progressed after the removal of his pituitary tumor. The patient is disease-free following ABVD/MOPP chemotherapy. A high level of steroids before treatment, albeit endogenous, did not preclude successful treatment of Hodgkin's disease. The effect of Cushing's syndrome on Hodgkin's disease has not been previously reported. PMID- 15111783 TI - Resolution of orbitocerebral aspergillosis during combination treatment with voriconazole and amphotericin plus adjunctive cytokine therapy. AB - Orbitocerebral aspergillosis has a very high fatality rate and cure is unusual. We describe the successful management of a child with cereberal aspergillosis who had a dramatic response to therapy with a combination of liposomal amphotericin and voriconazole with adjunctive cytokine therapy during immunosuppresive chemotherapy for acute lymphoblastic leukaemia. PMID- 15111784 TI - Good outcome for infant of mother treated with chemotherapy for ewing sarcoma at 25 to 30 weeks' gestation. AB - There have been only three previous reports of women with Ewing sarcoma who received chemotherapy while pregnant. A 17-year-old woman with Ewing sarcoma was treated with a combination of doxorubicin (Adriamycin) and ifosfamide during the 25th to 30th week of gestation, and the baby was delivered at the 32nd week. The baby was developing normally at follow-up at 8 months of age. This case report supports the idea that in this situation, the pregnancy can be continued to await fetal growth, and second-trimester chemotherapy may not have a deleterious effect on the fetus. PMID- 15111785 TI - Successful treatment of refractory autoimmune hemolytic anemia with monthly rituximab following nonmyeloablative stem cell transplantation for sickle cell disease. AB - Autoimmune hemolytic anemia (AIHA) can occur following hematopoietic stem cell transplantation (HSCT) and may be associated with other cytopenias. It can also occur in the context of chronic red cell transfusion in patients maintained on hypertransfusion regimens. There are an increasing number of reports on the successful treatment of autoimmune cytopenias with the monoclonal anti-CD20 antibody rituximab, including a few patients in a post-HSCT setting. The authors report the successful treatment with rituximab of refractory AIHA following allogeneic nonmyeloablative bone marrow transplantation in a child with sickle cell disease. PMID- 15111786 TI - Polycythemia vera in a child following treatment for acute lymphoblastic leukemia. AB - Polycythemia vera (PV), a hematologic stem cell disorder characterized by predominant erythroid proliferation, is extremely rare in childhood. Some PV patients develop acute leukemia, especially acute myelogenous leukemia, but cases of PV occurring after treatment of acute leukemia are rare. The authors describe a girl with an atrioventricular canal who was diagnosed with acute lymphoblastic leukemia (ALL) at 23 months of age, was cured with chemotherapy, and developed PV 7 years later. She went on to develop hepatic complications of PV that culminated in death from liver disease at 20 years of age, without recurrence of ALL. PMID- 15111787 TI - Early blastic transformation following complete cytogenetic response in a pediatric chronic myeloid leukemia patient treated with imatinib mesylate. AB - This article reports early blastic transformation of chronic myeloid leukemia (CML) in a child following a complete cytogenetic response induced by imatinib mesylate. A 14-year-old Japanese boy was diagnosed with t(9;22) cryptic CML in the chronic phase and treated with imatinib. His response to treatment was slow, but a major cytogenetic response was obtained at 142 days of therapy. However, he developed lymphoid blastic transformation at 9 months. He attained remission with acute lymphoblastic leukemia-type chemotherapy and then successfully received a non-T-cell-depleted allogeneic stem cell transplantation (allo-SCT) with his mother's two loci-mismatched donor cells. A sudden blastic transformation may occur even with a complete cytogenetic response induced by imatinib. CML patients who respond slowly to imatinib may still be candidates for allo-SCT, even when a major cytogenetic response is obtained. PMID- 15111788 TI - Stroke and elevated blood flow velocity in the anterior cerebral artery in sickle cell disease. AB - Transcranial Doppler ultrasonography provides a noninvasive method of predicting stroke risk in children with sickle cell disease. Elevated cerebral blood flow velocity in the terminal internal carotid or middle cerebral artery is associated with an increased stroke risk, but the clinical significance of elevated velocities in the other large intracranial vessels is unknown. The authors report stroke in two children with sickle cell disease and high blood flow velocity limited to the anterior cerebral artery. This suggests that elevated velocity in this vessel may be associated with an increased risk of stroke. PMID- 15111789 TI - Childhood acute myelomonocytic leukemia (AML-M4) presenting as catastrophic antiphospholipid antibody syndrome. AB - : The authors describe a 15-year-old girl presenting with a cerebral ischemic stroke as the first manifestation of catastrophic antiphospholipid antibody syndrome secondary to acute myeloid leukemia (AML). Despite treatment with anticoagulants, therapeutic plasma exchange, and chemotherapy, the patient developed multiorgan thromboses and failure, eventually culminating in death. This unusual presentation of AML has not been previously described in children. Clinical features of antiphospholipid antibody syndrome and current knowledge regarding its association with malignancies are reviewed. PMID- 15111790 TI - The surgical triumph at time zero and its full implications. PMID- 15111791 TI - The orbit after Converse: seeing what is not there. PMID- 15111792 TI - Clinical classification of positional plagiocephaly. AB - Positional plagiocephaly deformities have increased dramatically in all craniofacial clinics in the United States. There are multiple methods for evaluating the degree of deformity, all of which are expensive, time consuming, and have poor reproducibility. We present a clinical classification of plagiocephaly deformities that we have employed since 1998. The classification allows us to quantitate the degree of deformity in these children at any given time, to reliably determine quantitative changes from evaluation to evaluation. The technique is highly reproducible, cost effective and readily understandable to the family, as well as referring physicians. PMID- 15111793 TI - Pattern of the temporal branch of the facial nerve in the upper orbicularis oculi muscle. AB - The purpose of this study is to clarify a pattern of the temporal branch of the facial nerve in the upper orbicularis oculi muscle (OOM) and an impact in exploiting the frontalis myofascial advancement flap. The authors investigated the pattern of the temporal branch of the facial nerve in the upper OOM in 20 cadavers. The highest and lowest level of the nerve coursing into the OOM were measured at three different sagittal/vertical planes through the lateral canthus, midpalpebral fissure, and medial canthus, respectively. The authors designate a hazard zone that delineates a circle with 1.0-cm diameter and its center located inferiorly and laterally in the direction of -15 degrees 7.5 cm from the lateral canthus. The highest level of the those twigs that entered OOM on the X-axis and Y-axis with the origin of lateral canthus is +2.51 +/- 0.23 cm, +2.70 +/- 0.35 cm, and the lowest is 0 cm, +2.68 +/- 0.32 cm, respectively. The highest level of the those twigs on the Y-axis with the origin of lateral canthus, mid-palpebral fissure, and medial canthus is +3.47 +/- 0.27 cm, +3.49 +/- 0.45 cm, and +2.97 +/ 0.35 cm, and the lowest is +1.62 +/- 0.12 cm, +1.82 +/- 0.17 cm, and +1.63 +/- 0.22 cm, respectively. Those twigs of the temporal branch of the facial nerve coursed horizontally along the fibers of OOM with interconnections but did not cross over the superior orbital rim. The authors describe details of the temporal branch of the facial nerve in the OOM and designate a hazard zone, wherein the temporal branch should be spared. They also assure that injury of the temporal branch of the facial nerve is inevitable in the procedure of the frontalis myofascial advancement flap. PMID- 15111794 TI - Stoppers in RED II distraction device: is it possible to prevent pin migration? AB - Distraction osteogenesis has become popular for the treatment of hypoplastic congenital craniomaxillofacial anomalies. Rigid external distraction (RED II) after Le Fort III osteotomy was shown to be a highly effective treatment for the management of midface hypoplasia. This device is used with a halo vest, which is placed at the cranial equator. Intracranial penetration of the fixation pins of the halo is one of the complications of an external distraction device. To prevent pin penetration in rigid external distraction, the authors designed polylactic acid/polyglycolic acid (PLA/PGA) plates that were circular; the plates were 0.8 cm in diameter and had 1.5-mm holes in the center, through which the tip of the pins would pass. To quantify the applied torque by manual tightening of the screws of the distraction device and to measure intraosseous cone depth created by the penetration of the conical part of the screws with and without the PLA/PGA composite stopper, first an in vitro experiment was undertaken on cadaver. Then these PLA/PGA plaques, or stoppers, were placed over the bone surfaces of the cranium of the patients where the tip of the pins press. PLA/PGA stoppers are malleable and adapt their shape to the interactive forces between bone and the pins. They act as a second barrier, and spread the pressure of the screws to larger surfaces, thus securing better stabilization. The penetration of wider portions of the screw into scalp is reduced, minimizing the scalp damage caused by the screws. Biodegradable and biocompatible PLA/PGA stoppers avoid intracranial migration of the fixation pins, especially in children. PMID- 15111795 TI - Etiology and patterns of pediatric mandibular fractures in Portugal: a retrospective study of 10 years. AB - OBJECTIVE: To determine the pattern of occurrence of mandibular fractures in the pediatric population in Portugal. MATERIALS AND METHODS: This retrospective study reviews the records of patients 18 years of age or younger from the 10-year period 1993 to 2002. Age, gender, anatomic site, cause of the accident, weekly and monthly variation, location and type of fractures, presence and location of associated injuries, treatment methods, and complications were reviewed. RESULTS: During this 10-year period, 521 patients with 681 mandibular fractures were treated. Motor-vehicle accident (MVA) was the most common (53.9% patients) cause of fracture. Almost half of the patients (48.8%) were in the oldest age group (16 to 18 years old). The condyle of the mandible was involved in 31.0% of the fractures. Maxillomandibular (MMF) fixation was used in 534 (78.4%) fractures. Overall mortality in this series was 0.6% (3 patients); mortality was caused by multiple traumas, mainly head trauma. CONCLUSION: There is a need to reinforce legislation aimed to prevent MVA and the total enforcement of existing laws to reduce maxillofacial injuries among children and adolescents. PMID- 15111796 TI - Craniofacial abnormalities in a murine knock-out model of mucopolysaccharidosis I H: a computed tomography and anatomic study. AB - The genetic mucopolysaccharidoses are a group of lysosomal storage diseases in which mucopolysaccharides (glycosaminoglycans) accumulate as the result of a malfunction or lack of a lysosomal degradation enzyme. There are currently seven known forms of mucopolysaccharidoses. Type I results from an enzymatic deficiency of alpha-L-iduronidase. There are three subtypes of mucopolysaccharidoses I that are commonly recognized: Hurler syndrome, Hurler-Scheie syndrome, and Scheie syndrome. Of the three subtypes, Hurler syndrome has the most severe clinical picture. Craniofacial anomalies and cognitive impairment are some of the more pronounced features of Hurler syndrome. Hurler syndrome has been described in cats, dogs, mice, and human beings and is inherited as an autosomal recessive trait. The biochemical nature of the disease is preserved across species lines. Clinically, the disease has similar effects in human beings and animals. It has been difficult to reverse the phenotype of the disease even with replacement of the defective alpha-L-iduronidase enzyme. The purpose of this study is to characterize the cranio-facial differences in the murine knock-out model of Hurler syndrome objectively. Twenty-three measurements were taken from computed tomographic scans in a coronal and sagittal plane on 24 black C57/B6 knock-out Hurler syndrome mice. The seven statistically significant measurements are width of the cervical canal, height of the foramen magnum, width between the external auditory canals, width of the skull base at the mandibular condyles, midocular distance, spread of the mandibular condyles, and width of the zygoma at the maxilla. This information now provides researchers with objective data from living Hurler syndrome-affected mice that will allow them to follow therapies directed at improving craniofacial outcomes for any therapy over time. PMID- 15111797 TI - Reliability and validity of a modified lateral cephalometric analysis for evaluation of craniofacial morphology and growth in patients with clefts. AB - In previous intercenter studies on craniofacial morphology in patients with unilateral cleft lip and palate, probable surgical-induced changes in mandibular morphology and spatial position related to posterior vertical maxillary morphology were identified by our group. These changes could not be detected in other cephalometric cleft studies because posterior vertical maxillary height and vertical mandibular ramus length were not measured simultaneously. This study presents a modified digital lateral cephalometric hard and soft tissue analysis (Onyx Ceph software, version 2.5.6.; Image Instruments GmbH, Chemnitz, Germany) to evaluate craniofacial morphology and growth patterns in patients with clefts. Forty controls without clefts were used to evaluate the accuracy, reliability, and validity of this analysis for future cleft research. Measurement error according to the method of Bland and Altman was less than 1.00 degrees and 1.00 mm, whereas squared correlation coefficients (r) according to the method of Sackett et al showed a high reliability. Method comparison tests according to the method of Bland and Altman clearly showed that the modified digital cephalometric analysis ("test") was valid for future cleft research compared with the "gold standard" (conventional cephalometry). PMID- 15111799 TI - Biomechanical analysis of hydroxyapatite cement cranioplasty. AB - A recent review of the authors' experience with hydroxyapatite (HA) cement cranioplasties revealed a high infection rate. During removal of these implants, all were loose and fractured. Forty percent of these patients had a history of minor trauma at the site of cranioplasty before experiencing infection. Minor trauma may fracture HA cranioplasties and result in infection. The purpose of this study is to determine the force to fracture full- and partial-thickness cranial defects reconstructed with HA cement and to compare peak loads of differing HA cement cranioplasty techniques. Standardized craniotomy defects were created in five fresh cadaver heads. Full-thickness defects were reconstructed with either rigid or flexible titanium mesh and then covered with HA cement. Partial-thickness defects were reconstructed with HA alone. After setting, a uniaxial impact was delivered to each of the defects. Peak loads were recorded, and defects were examined for evidence of fracture.Predictable fractures of the HA cranioplasties occurred at 1200 N in all full-thickness defects reconstructed with mesh and a thin layer of HA. Implant loosening and chipping was similar to what was seen clinically in the authors' patients with infections. Full-thickness defects in which titanium mesh was shaped like a cup and filled with a thick layer of HA resist fracture at 1200 N. Partial-thickness defects reconstructed with HA alone also do not fracture at this peak load. Patient selection, defect characteristics, and reconstructive techniques are factors that need to be considered before using HA cement for cranioplasty purposes. PMID- 15111801 TI - Intermittent parathyroid hormone treatment enhances guided bone regeneration in rat calvarial bone defects. AB - This study investigates the effects of intermittent parathyroid hormone (PTH(1 34)) treatment on bone regeneration and mechanical strength of critically sized rat calvarial bone defects covered with expanded membranes. A full-thickness bone defect (diameter 5 mm) was trephined in the central part of the parietal bones in 20-month-old female Wistar rats. The bone defects were covered with an exocranial and an endocranial expanded polytetrafluoroethylene membrane. The animals were killed 35 days after operation. 60 microg PTH(1-34)/kg was administered daily during the healing period, and control animals with calvarial bone defects were given vehicle. Mechanical testing was performed by a punch out testing procedure by placing a steel punch (diameter 3.5 mm) in the center of the healed defect. After mechanical testing, the newly formed tissue inside the defect was removed and the dry weight and ash weight were measured. PTH(1-34) increased dry weight by 48%, ash weight by 51%, and ash concentration by 26%. PTH(1-34) also augmented the mechanical strength of the new bone formed inside the defect by increasing ultimate stiffness by 87%. No differences in body weight were found between the vehicle-injected and the PTH-treated animals during the experiment. The experiment demonstrates that intermittent PTH(1-34) treatment increases bone deposition and enhances mechanical strength of healing rat calvarial defects covered with expanded polytetrafluoroethylene membranes. PMID- 15111803 TI - Aberrant bony vasculature associated with activating fibroblast growth factor receptor mutations accompanying Crouzon syndrome. AB - Fibroblast growth factor receptor mutations are associated with and, in fact, cause most syndromes presenting with craniosynostosis. This knowledge has resulted in a shift in the paradigm of suture fusion causation; it was thought previously that abnormal tensional forces arising in the cranial base caused fusion of the vault sutures, but it is now understood that aberrant intercellular signaling in the developing skull leads to abnormal suture morphogenesis. Although the mutations associated with these syndromes are known and the phenotypic consequences are well documented, the pathway from mutation to phenotype has yet to be elucidated. Surgical reconstruction is the primary treatment of craniofacial abnormalities associated with craniosynostotic syndromes such as Crouzon syndrome. In many cases, calvarial vault reshaping is dependent on the quality of the autologous bone available; however, the bone of patients with craniosynostosis syndrome is often more brittle, thinner, and less robust than cranial bone from nonaffected donors. The relation between syndromic craniosynostoses and this bone has not been previously described. In this study, the osteon and blood vessel diameters of calvarial bone from patients with Crouzon syndrome and age- and sex-matched normal calvarial bone are measured. Statistical analysis demonstrates a quantitative and significant difference in the blood vessel diameter but not in the osteon diameter. This finding could be a result of abnormal blood vessel development caused by the fibroblast growth factor receptor mutation occurring before and coincident with bone formation and leading to weakened and fragile bone tissue. PMID- 15111806 TI - Basic studies on the bone formation ability by platelet rich plasma in rabbits. AB - PURPOSE: Platelets, which contain many growth factors, such as platelet-derived growth factor (PDGF) and transforming growth factor-beta(TGF-beta), can be obtained in high concentrations by centrifugal separation and are being used as platelet-rich plasma (PRP) in clinical applications. The authors evaluated the bone formation ability by PRP in rabbits. MATERIALS AND METHODS: In experiment 1, the authors made platelet precipitations from the whole blood obtained from rabbits. Each precipitated platelet was resuspended in fibrin glue. The glue was applied to a 1-mm wide slit defect of the same rabbit mandible. After 1 week, the authors observed the histologic appearances. In experiment 2, the fibrin glue containing the precipitated platelet was applied to a bicortical defect of a rabbit cranium with beta-tricalcium phosphate (beta-TCP) granules as artificial bone material. After 1 week, 1 month, and 2 months, the authors observed the radiologic and histologic appearances. RESULTS AND CONCLUSIONS: In both experiments, a prosperous bone formation was observed from the beginning more in the group treated with PRP than in the group untreated. In experiment 2, gradual appearances of the bone formation were observed in the bubble of beta-TCP. In application with artificial bone materials, bone formations by PRP were suitably observed. PMID- 15111807 TI - Transmandibular K-wire in the management of airway obstruction in Pierre Robin sequence. AB - The Pierre Robin sequence was first described by Pierre Robin in 1923 as a triad of micrognathia, U-shaped cleft palate, and glossoptosis. Although the problems associated with Pierre Robin sequence may be numerous, the most acute problem in affected newborns is upper airway obstruction. The causes of upper airway obstruction are heterogeneous, and treatment is controversial and may include a long stay in an intensive care setting. Most patients could be treated conservatively with prone/lateral positioning and close clinical observation. A more aggressive approach is to hold the tongue forward surgically by a lip-tongue adhesion (glossopexy) technique, and if all else fails, it might be necessary to perform a tracheostomy. The purpose of this article is to report an unconventional technique for the management of airway obstruction in Pierre Robin sequence. A transmandibular K-wire was used in two patients with Pierre Robin sequence to prevent airway obstruction. The author was not the surgeon who placed the K-wire, and in one of the patients, the K-wire was retained for 4 years before being removed by the author. Both patients had surgical intervention to manage the airway problem in the form of lip-tongue adhesion in addition to the transmandibular K-wire. In conclusion, the value of using a transmandibular K wire in the two cases presented here could not be determined and was questionable. PMID- 15111808 TI - Isolated supraorbital rim fracture displaced out of scalp: a case report. AB - Isolated fractures of the supraorbital rim are rarely seen. In this report, an isolated supraorbital rim fracture is presented in which the segment of the rim was completely removed from its location onto the frontal bone in a traffic accident. It was thought to be a foreign body and was delivered outside the scalp in the emergency service. It was replaced 24 hours after the trauma, and the patient healed without any problem. PMID- 15111809 TI - The transconjunctival approach for orbital bony surgery: in which cases should it be used? AB - The advantages and disadvantages of the transconjunctival approach were examined to determine its indication for orbital bony surgery. The transconjunctival approach was used in 22 patients. The average follow-up was 13 months. Two patients had an intraoperative lower eyelid laceration because of excessive traction. Lower eyelid retraction occurred in 5 patients after surgery, although only 1 of them required surgical repair. With a transconjunctival approach alone, the exposure of the orbital lateral wall is limited and incorporation of a lateral incision has been found to be necessary. Postoperative eyelid retraction seems to occur even in the transconjunctival approach. The transconjunctival approach is best indicated in cases with an orbital medial wall fracture because it provides much easier access than any cutaneous approach. PMID- 15111810 TI - Sphenoidal cerebrospinal fluid rhinorrhea: an original surgical approach. AB - Management of patients with sphenoidal cerebrospinal fluid rhinorrhea remains controversial. With consideration of the etiology of cerebrospinal fluid leaks, size of the sinus, location of the fistula, and previous surgery, one may choose either an extracranial or transcranial approach. In two patients in whom conventional procedures failed or seemed to be inapplicable, the cerebrospinal fluid leak was successfully treated through an original transfacial approach with palpebral incisions. A transzygomatic osteoplastic flap provided access to close the fistula and to exclude the sinus with a bone graft. PMID- 15111811 TI - Occlusal difficulties after simultaneous mandibular and maxillary distraction in an adult case of hemifacial microsomia. AB - A 23-year-old woman with hemifacial microsomia type IIB was treated by simultaneous mandibular and maxillary distraction. During the course of the distraction, cephalometric analysis showed that the maxilla was not moving downward and that the upper edge of the mandibular condyle was moving upward into the space between the condyle and the glenoid fossa. This phenomenon could lead to early consolidation of the osteotomized maxilla, resulting in malposition and occlusal difficulties. This is thought to be a problem unique to bimaxillary distraction of hemifacial microsomia type IIB with severe hypoplasia of the mandibular ramus and space between the condyle and the glenoid fossa. Surgeons should be alerted to this risk and prepared to address it when performing this procedure. PMID- 15111812 TI - New method of pediatric cranioplasty for skull defect utilizing polylactic acid absorbable plates and carbonated apatite bone cement. AB - Cranial defect repair in the pediatric population requires a variety of special considerations. The pediatric skull has a dynamic nature that prohibits the use of rigid fixation, which is commonly applied in the adult population. A technique using a combination of polylactic acid plates and carbonated apatite bone cement has been devised by our group. Skull defects of varying sizes were repaired in 34 pediatric patients. Patients were examined on postoperative day 3 and at 3 months via three-dimensional computed tomography scans. Patients have been followed up to 60 months after surgery without complications or failures to date. This method benefits the pediatric patients undergoing cranioplasty by minimizing the insertion of long-term foreign bodies and allows the possibility for transformation of this construct into viable tissue. PMID- 15111813 TI - Case report: endoscopic excision of a nasoglabellar dermoid. AB - Endoscopic techniques have greatly reduced the morbidity of excising benign orbitofacial masses. Access, visualization, and dissection of these masses are comparable to open approaches without the use of large or conspicuous incisions. Existing reports focus on treatment of dermoids and other benign masses in the forehead, brow, and temple. We describe the successful endoscopic excision of a nasoglabellar dermoid, which is much less common. PMID- 15111814 TI - Augmentation by autologous adipose tissue in cleft lip and nose. Final esthetic touches in clefts: part I. AB - Although patients with a cleft lip and palate undergo many surgeries and other therapeutic procedures in the course of their treatment, many are still deeply concerned with their handicap and continue searching for perfection in their appearance. Augmentation using the subject's own fat cells involves minor invasion, is readily available, is an unpretentious method regarding time and cost, and has no contraindications. This method can serve to supplement a hypotrophic scarred upper lip and nasal columella, and by improving the volume, it induces a more natural contour, which reduces the stigmatizing deformity as well as the visibility of externally apparent scars. Using this approach, five patients with a complete cleft have been treated. The median follow-up interval is 22 months (through January 2003). The procedure and postoperative course had a pleasing outcome and were without any complications. The disadvantage was the temporary effect of the outcome, which necessitated repeated application every 7 months on average. PMID- 15111816 TI - The use of Novabone and Norian in cranioplasty: a comparative study. AB - Bone replacement products have enhanced the ease of reconstructing bone while improving morbidity related to bone harvest. Although these products are successfully used, studies of bone healing and biomechanical strength are lacking. We aimed to compare how Norian CRS (cranial replacement substance) and Novabone C/M heal in a cranial defect. Adult New Zealand rabbits underwent removal of a critical size cranial defect. The defect was filled with Novabone (n = 8), Novabone plus demineralized bone matrix (n = 8), or Norian (n = 8), or it was left empty (n = 8). Rabbits were euthanized at 8 weeks. Cranial specimens were harvested and soft radiographs, contact microradiographs, and biomechanical testing were done. Soft radiographs revealed opacification like adjacent bone with Novabone, which was augmented when Novabone was combined with demineralized bone matrix. Norian maintained an opaque appearance. The control group did not heal. Contact microradiographs demonstrated bone within the healing defect with Novabone, which was augmented by demineralized bone matrix. Norian was not replaced with bone but served as a scaffold for bone formation. Biomechanical indentation testing demonstrated that the stiffness of Norian was the highest. Novabone plus demineralized bone matrix had a higher stiffness than Novabone alone. All experimental groups had a statistically significant difference compared with Norian. None of the groups achieved the strength of unoperated native bone. Studying two popular products, we found evidence that Novabone was incorporated into cranial bone, regenerating the bone. Novabone healed at a faster rate, creating a stronger product, with demineralized bone matrix. The biomechanical strength of the healed defect was higher in the Norian group, because the bone cement remained solid and was not incorporated, unlike crania reconstructed with Novabone. PMID- 15111817 TI - Anatomical consideration of the congenital nasal pyriform aperture stenosis: localized dysostosis without interorbital hypoplasia. AB - Congenital nasal pyriform aperture stenosis may be isolated or associated with other midline anomalies. The aim of the study was to describe the measurements and features of the interorbital structure and midface in congenital nasal pyriform aperture stenosis. The computed tomography scans of eight patients (two girls and six boys) were retrospectively reviewed. Several distances were obtained at the orbital and midface levels and compared with normative data. The average width of the pyriform aperture was 5.5 mm +/- 1.6, and there was overgrowth of the pyriform aperture rim (nasal process of the maxilla). No skeletal anomalies were present other than the congenital nasal pyriform aperture stenosis. It was associated with a single median maxillary central incisor in two cases. The anterior and lateral interorbital distances were normal (17.4 mm and 65.4 mm, respectively). The midface was not hypoplastic. There was no brain malformation. Congenital nasal pyriform aperture stenosis is a localized dysostosis of the pyriform aperture rim without interorbital or midface hypoplasia. It may be associated with a single median maxillary central incisor but cannot be viewed as a minor form of holoprosencephaly. PMID- 15111818 TI - Glomus tumors: therapeutic role of selective embolization. AB - The primary goals of preoperative embolization of glomus tumors are to reduce the blood loss in the surgical field, minimize the risk of operative complications, and prevent recurrence by contributing to complete resection. Obliteration of a vascular channel may provide additional help in symptomatic relief by decreasing the tumor volume. Selective transarterial embolotherapy outcomes of 17 patients, most of whom had inoperable tumors and were unable to have surgery/radiotherapy because of poor general health, were evaluated to investigate the therapeutic contributions, efficacy, and safety of embolization techniques in the treatment of different glomus tumors and to examine the role of the embolotherapy in the treatment algorithm of such lesions. The pattern of vascular supply of these tumors was also documented. Eleven glomus jugulotympanicum, 4 glomus caroticum, and 2 glomus vagale tumors were embolized for palliative or curative purposes between 1992 and 2000. Coils and combination of coils plus polyvinyl alcohol were used for embolization. Relief of symptoms and patient satisfaction were analyzed within 3 months after the intervention by a questionnaire combined with full ear, nose, and throat and neurological examinations. The results indicate that the use of embolotherapy in the treatment of glomus tumors must be basically preoperative but is not curative. PMID- 15111819 TI - Benign osteoma with Gardner syndrome: review of the literature and report of a case. AB - Gardner syndrome, a variant of familial adenomatous polyposis, is an autosomal dominant disease characterized by gastrointestinal polyps that develop in the colon as well as in the stomach and upper intestine (duodenum), multiple osteomas, and skin and soft tissue tumors. Cutaneous findings include epidermoid cysts, desmoid tumors, and other benign tumors. Polyps have a 100% risk of undergoing malignant transformation; consequently, early identification and therapy of the disease are critical. Osteoma is a benign neoplasm of bone tissue that is characterized by slow continuous growth and is the most common accompanying bone lesion seen in Gardner syndrome. The authors report a case of Gardner syndrome that was operated on because of the mandibular osteoma. PMID- 15111820 TI - Visual, auditory, and tactile temporal processing in children with oral clefts. AB - Neurocognitive disorders may compromise the outcome of surgical cleft lip palate repair and thus need to be identified. Processing of rapidly changing sequential information (temporal processing) is a fundamental neurocognitive capacity that may contribute to various communication functions and has been found impaired in several developmental disorders. The occurrence of temporal processing difficulties in the cleft population is not known, however. We investigated the relation between oral clefting and temporal estimations of simultaneity/nonsimultaneity in visual, auditory, and tactile modalities. Subjects were 10-year-old controls and children with oral clefts of various types [i.e., cleft lip (alveolar), cleft lip and palate, cleft palate, and cleft palate submucous]. The visual and tactile tasks but not so clearly the auditory task differentiated the groups. Further, paralleling previous findings, the patients with cleft lip and palate outperformed the others, whereas the cognitive temporal processing acuity of the cleft palate and cleft palate submucous children was worse. PMID- 15111821 TI - Early failure of absorbable plating in a patient with syndromic brachycephaly. AB - The use of bioabsorbable plating systems for rigid fixation after cranial remodeling surgery has become the standard of care in the treatment of syndromic and nonsyndromic craniosynostoses at pediatric craniofacial centers in North America. The advantages of these absorbable plating systems over metallic plates and screws have been well documented. The absorbable plates have been used with remarkable safety and efficacy for more than 15 years. An unusual case of repeated early failure of an absorbable plating system in a patient with syndromic brachycephaly complicated by the development of hydrocephalus is reported. The reoperative rates for cranial remodeling surgery at our institution and the possible causes for this failure are discussed. PMID- 15111823 TI - Hypophosphaturic mesenchymal tumor of the ethmoid associated with oncogenic osteomalacia. AB - Oncogenic osteomalacia is an uncommon syndrome characterized by bone pain, proximal muscle weakness, hypophosphatemia, hyperphosphaturia, and a low plasma concentration of 1,25-dihydroxy-vitamin D. The disease affects both sexes at around 40 years of age, although it can sometimes affect children and adolescents. Generally, the syndrome is associated with a tumor, usually benign, of mesenchymal origin and is resolved after removal of the tumor; this syndrome can sometimes be associated with malignant tumors. These tumors seem to be histologically heterogeneous and are generally localized in soft tissues and bone. In this article, a case of oncogenic osteomalacia associated with a hypophosphaturic mesenchymal tumor of the ethmoid is reported in a 24-year-old man. After surgical and radical removal of the tumor, the patient noted a decrease in the clinical symptoms and signs. PMID- 15111824 TI - Soft tissue facial anthropometry in Down syndrome subjects. AB - The three-dimensional coordinates of 50 selected soft tissue facial landmarks were digitized on 28 white Italian subjects with Down syndrome (17 male and 11 female subjects aged 12 to 45 years) and 429 healthy controls of comparable ages by an electromechanical instrument. From the landmarks, 16 facial dimensions were calculated. Data were compared with those collected in healthy individuals by computing z-scores. Overall, most variables were smaller in subjects with Down syndrome than in their normal controls selected for sex, age, and ethnicity (negative z-scores), even if not all of them reached statistical significance. Independently of sex, subjects with Down syndrome had faces that were significantly (P <0.05, paired Student t test) narrower (skull base and mandible), less deep (upper, middle, and lower face), and shorter (face and nose height) than the faces of normal subjects. Additionally, ear width and length were significantly reduced on both sides of the face. Only facial height was significantly different between sexes (P = 0.023, unpaired Student t test), with a female z-score that was more than two times the relevant male value. The present investigation represents the first detailed quantitative analysis of the facial soft tissue characteristics of Italian white subjects with Down syndrome. PMID- 15111825 TI - Highlights of the proceedings from the 10th International Congress of the International Society of Craniofacial Surgery. PMID- 15111826 TI - [Laparoscopic adrenalectomy in multiple endocrine tumors, in secreting and non secreting lesions]. AB - AIM: Personal experience in laparoscopic adrenalectomies (LA) for secreting and non-secreting tumors is presented. METHODS: Between March 1995 and December 2001 a total of 111 LA (58 left, 49 right and 4 bilateral) were performed in 60 females and 51 males, mean age 47.5 (range 8-81) years, for: 38 Conn diseases, 24 incidentalomas, 15 pheochromocytomas, 13 Cushing diseases, 4 kysts, 3 angiomyolipomas, 1 adreno-genital syndrome, 1 hydatidosis, 1 hyperplasia, 1 ganglioneuroma, 1 oncocyte adenoma, 1 adrenal fibrous tumor, 4 cortical carcinomas and 4 metastases (from renal carcinoma, breast carcinoma, leiomyosarcoma and rabdoid sarcoma, respectively). In all cases LA was transabdominal with a lateral flank approach. RESULTS: Mean global operative time was 88.6 minutes (range 35-240). Conversion rate was 1.8% (2/111). There was low postoperative pain. Mean hospital stay was 4.3 days (range 2-13). There were 0.9% (1/111) 30-day mortality and 4.5% (5/111) morbidity. During a mean follow-up of 41 months (range 1-81), the 67 secreting patients were disease-free. Concerning 8 malignant cases, mean follow-up was 40.5 months (range 9-72) with 3 cortical carcinomas disease-free and 1 dead for stroke, 2 metastases (1 leiomyosarcoma and 1 breast carcinoma) dead for disease and 2 disease-free. There was no port-site metastases. CONCLUSIONS: LA seems safe and effective when performed in experienced Centers on endocrine surgery and laparoscopy. PMID- 15111827 TI - Surgical treatment of liver metastases from breast cancer. AB - BACKGROUND: The prognosis of patients with hepatic metastases (HM) from breast cancer receiving no treatment is extremely poor. The aim of this study is to report results of liver resections for breast cancer liver metastases. METHODS: Retrospective analysis of 17 women who underwent hepatic metastasectomy with curative intent in the Centre Leon-Berard in Lyon and in the Department of General Surgery of University of Genoa. RESULTS: The mean number of HM was 2.2 (range: 1-9). The following resections were performed: extended hepatectomies (7), hepatectomies (5), lobectomies (2) and segmentectomies (3). The median number of resected segments was 3 (range: 1-5). Perioperative complications occurred in only 1 patient (hepatic venous hypertension). Postoperative complications were seen in 5 patients: pleural effusion (3), pneumonia and fever (1) and biliary fistula (1). Nine patients developed recurrence disease: 6 patients died and 3 underwent a 2nd hepatic resection. One-year survival after hepatic resection was 92%, 2-year survival was 85%, 3-year survival was 52% and 5 year survival was 41% (Kaplan-Meier estimate). CONCLUSIONS: Our data, although limited, suggest that liver resection is a viable treatment option for selected patients that can be performed safely. It should be considered in individual patients if the operative risk is low, if no extrahepatic disease is present and provided a complete resection with clear margins is technically feasible. PMID- 15111828 TI - [Common bile duct stones: the surgical treatment is always valid]. AB - AIM: The treatment of common bile duct stones has changed with the new therapeutic techniques, that have replaced the conventional therapy, represented by surgery. Anyway, they could cause some problems, that must be regarded. Therefore, we wish to confirm the importance of the conventional surgery in the management of patients with common bile duct stones. METHODS: A total of 147 patients were operated for common bile duct stones (73 in emergency and 74 in election). The intraoperative cholangiography was carried out in 141 patients and a choledocoscopy in 130 patients. A drain of Kehr was positioned in 120 patients, a bilio-digestive anastomosis in 26 cases and in 1 case there was a direct suture of the common bile duct without drain. All patients were treated with a short term antibioticotherapy, protracted to 5 days in the emergency cases. In the patients with the drain of Kehr there was a control cholangiography after 7 days from operation and it was removed after 25 days. The analysis of the results was done dividing the patients according to the age: <75 years old and >75 years old. RESULTS: Nobody died during the operation. Complications were 17.4% in the patients >75 years old and 2,6% in the patients <75 years old. There were 2 death, in the postoperative period, for the group >75 years old. Cases operated in emergency were 68.1% of "old" patients and 37.2% of "young" patients. The postoperative period was 14.9+/-9.2 days for the "old" group and 10.9+/-5.2 days for the "young" group. CONCLUSIONS: The treatment of common bile duct stones is still a surgical treatment, particularly for "old" patients. PMID- 15111829 TI - Quality of life in patients with an ileostomy. AB - AIM: Temporary or permanent ileostomy is a common procedure in colorectal surgery. Our aim was to assess the impact of ileostomy on quality of life and the relevant risk factors. METHODS: A 33-question questionnaire was sent, by mail, to 74 patients (over 70, between 70 and 50 and under 50 y old). The questionnaire explored 5 parameters of function: systemic symptoms, bowel symptoms, functional impairment, social impairment and emotional impairment. Further questions investigated the need for a nurse or relative to assist with management of the stoma, the frequency of changing the bag and the appliance, diet and the ability of patients to attend their normal daily activity. RESULTS: Thirty-four patients completed the questionnaire. The final quality of life score was similar for all the 3 age groups but elderly patients needed more assistance in the stoma management. The quality of life scores in males patients were significantly better than in females. No statistically significant difference was observed comparing loop ileostomy versus end ileostomy and Crohn's disease versus ulcerative colitis. CONCLUSIONS: Quality of life is not influenced by age, underlying inflammatory disease or type of ileostomy. Males report less impact on quality of life than women. PMID- 15111830 TI - [Mini-invasive treatment of pectus excavatum in adolescence. Initial experience]. AB - BACKGROUND: Personal preliminary experience with Minimally Invasive Repair of Pectus Excavatum (MIRPE), "Nuss" procedure, using VATS is reported. METHODS: From January 2001 to February 2002, MIRPE has been performed on 5 patients (age range 13-18 y; mean 14.8 y). Under general anesthesia, a curved steel bar is inserted into the retrosternal tunnel between 2 bilateral midaxillary line incisions. The tunnel passes initially under the pectoral muscles and enters the pleural space at level of the mammilary line. Under thoracoscopic vision, the bar is passed through the tunnel with the concavity facing the front and then is turned over thereby correcting deformity. An epidural catheter relieved perioperative pain successfully. RESULTS: In all patients the repair has been good. Mean hospital length of stay has been 6.8 d. Pneumothorax occurred in 1 patient requiring tube thoracostomy. After 45 d 1 patient had a bar displacement requiring a reoperation. All patients have a normal life. CONCLUSIONS: The Minimally Invasive Repair of Pectus Excavatum is an effective procedure even in adolescence. Thoracoscopic vision makes safer the creation of the retrosternal tunnel and the passage of the bar. Short-term results have been good. Further follow-up is necessary to determine long-term results. PMID- 15111831 TI - [Indications, techniques, complications and results of sleeve lobectomy]. AB - AIM: The authors analyse their personal series and compare it with reported data in order to assess the functional results sleeve lobectomy offers with respect to traditional techniques. METHODS: Sixty-six sleeve lobectomies were carried out from 1986 to 2000. In 49 cases the operation was carried out on the basis of criteria of choice and in 17 of functional necessity. Three vascular sleeves and 5 tangential vascular plastic operations were associated. The disparity of lumen was corrected with oblique sections on the intermediate bronchus according to Merendino. In the case of reimplantation of the intermediate on the left main bronchus high frequency jet ventilation was necessary. The anatomoses were covered with broad pedunculated pleural strips. In 3 cases extramucosal myectomy of the medio-thoracic esophagus was carried out associated in 1 case with mucous resection and subsequent direct suture. RESULTS: Morbility was 34.5% (21% atrial fibrillation, 7.5% parenchymal complications, 4.5% anastomotic complications and 1.5% mediastinitis). Mortality at 30 days was 6% (massive hemoptysis in 3 cases and respiratory failure in 1 case). The pTNM of the 57 survivors was IB in 35 cases, IIB in 6 and IIIA in 16. Follow-up showed 5-year survival of 62% for stage IB and 24% for stage IIIA. CONCLUSIONS: On the basis of their experience, with a postoperative mortality and a rate of complications comparable to literature data, the authors consider that sleeve lobectomy, although it presents postoperative problems requiring more prolonged clinical control, is preferable to traditional operations because of the functional advantages it offers patients with lung cancer. PMID- 15111832 TI - [Postoperative pneumonia caused by Candida in 500 lung surgery patients]. AB - BACKGROUND: Postoperative infections continue to be one of the most frequent complications in hospitalized patients. The incidence of fungal infection has been steadily rising. While Candida albicans remains the most common yeast species isolated in hospitalized patients, other Candida species have been increasingly isolated. METHODS: From 1996 to 2001, a prospective study of 500 consecutive lung surgery patients treated by the same surgical team was conducted to monitor the number of postoperative infections and to evaluate the epidemiology of bacterial and fungal infections. RESULTS: At least one postoperative infection developed in 18% of patients. Of the 51 patients who developed postoperative pneumonia, 24 underwent microbiological examination of sputum. In 19 of them, the culture yielded isolates of one or more Candida species. Slightly under half of patients (47%) with cultures positive for mycetes received treatment with fluconazole until the clinical infection resolved. The drug was well tolerated by all treated patients. CONCLUSIONS: The incidence of fungal infection has increased also at our center. In 19 of 24 patients with postoperative pneumonia, sputum cultures yielded Candida species isolates. Many factors may be contributing to the rise in fungal infections after surgery. As pneumonia caused by Candida led to a significantly longer length of hospital stay in our case series, we draw attention to the importance of early diagnosis of postoperative mycotic lung disease in order to institute timely and targeted therapy. PMID- 15111833 TI - [Venous microanastomoses by extraluminal cuff versus standard technique]. AB - AIM: Venous microanastomoses are more difficult to carry out in comparison with the arterial ones, because of the characteristics of the vascular wall. The suture with loose stitches is the usual surgical technique, but it has 2 disadvantages: a long time of execution and the presence of foreign material in the anastomosis. To avoid these complications, we an extraluminal silicone cuff has been used. METHODS: We performed 70 microanastomoses on the internal jugular vein of Sprague-Dawley rats with these 2 techniques, estimating the immediate and late permeability and postoperative complications. Moreover, a histological study of all the anastomoses was carried out on the 15th day and after 1 mo from the intervention. RESULTS: The mean time requested to perform the microanastomosis is significatively shorter when using the extraluminal cuff. The late permeability with standard suture is 97% and 77% with the cuff. The number of complications with standard suture is significatively lower than with cuff, where rates of 20% of dehiscence and 14% of trombosis were observed. The histological study showed a poor alteration of the vascular wall, with important subendothelial hypertrophy on cuff microanastomosis and a great permanent histopatologic alteration if the standard technique had been used. CONCLUSIONS: The suture with loose stiches is a very good technique to use for this type of microanastomosis, even if the extraluminal cuff can be used in microanastomosis of a free graft or when it is necessary to perform more than 1 microanastomosis. PMID- 15111834 TI - [The surgical needs of elderly patients in day-surgery]. AB - AIM: The possibility of carrying out surgery in day-surgery (DS) conditions is gradually becoming reality in most branches of surgery; in recent years, DS has also found a place in general surgery, with unquestionable advantages for the management of patients, particularly the elderly. The purpose of the present study is to investigate general surgery needs in DS conditions in elderly patients in order to analyse the clinical-administrative feasibility of DS procedure and the level of its acceptability in this group of patients. METHODS: A specially drafted questionnaire was submitted to patients admitted over a period of 18 months to the General Surgery Division of the University of L'Aquila. The details investigated in the questionnaire were: age, sex, educational qualification, working activity (independent or employee), presence in the patient's relational entourage of health workers willing to serve the patient at home, the patient's willingness to undergo the surgery for which he was admitted to the DS. The questionnaires of the over-64s were examined and the percentage of patients willing to undertake DS intervention instead of normal hospitalisation was evaluated; this willingness was then compared with the parameters sex, educational qualification, working activity and presence in the patient's relational entourage of health workers willing to assist the patient at home. RESULTS: 317 questionnaires were compiled. In 78 cases (24.6%) the patient was older than 64; the data for these patients were extrapolated. Willingness to undergo surgery in DS conditions was expressed by 23 patients (29%), whereas 55 patients (71%) stated that they were not available. In relation to sex, willingness to use the DS was 29.7% in the 37 male patients and 29.2% in the 41 females. With reference to the qualification, willingness was expressed by 71.4% of patients with a degree, 26.9% of patients with a high school leaving certificate and 15.7% of patients with a lower qualification. DS-willingness was 28% in the 75 pensioners and 66.6% in the 3 patients who worked for themselves. As regards the presence in the relational entourage of the patient of health workers ready to assist at home, DS-willingness was 23.9% in the 71 patients who did not have anyone in their relational entourage and 85.7% in the 7 patients who had such a health worker in their entourage. CONCLUSIONS: Willingness to undergo DS by elderly patients is mainly influenced by three factors: educational qualification, working activity and possibility to have home assistance from someone in the relational entourage; these parameters can be considered veritable selection criteria for DS in the elderly. The quality improvement in assistance levels and the introduction in the near future of telematic communication system could lead to an extension of DS indications to the elderly. PMID- 15111835 TI - [Multiple endocrine neoplasia type 2. Biological and clinical aspects]. AB - CLASSIFICATION: Medullary thyroid carcinoma (MCT), a rare thyroid malignancy originating from the parafollicular C cell, may occur either as a hereditary or a non-hereditary entity. Hereditary MCT can occur either alone, familial MCT (FMCT), or in multiple endocrine neoplasia type 2 (MEN 2), associated with other endocrinopathies such as pheochromocytoma and/or hyperparathyroidism (MEN 2A and 2B). These hereditary disorders are due to germline mutation in the RET proto oncogene. Early diagnosis and treatment significantly improve the outcome of patients with MCT. DIAGNOSIS: In hereditary MTC, the MTC is usually multifocal and bilateral. Serum calcitonin measurement, a marker of disease, is superior to fine needle aspiration cytology in suggesting the diagnosis of MCT. Other investigations including ultrasonography, chest X-ray, computerized tomography and MRI may provide valuable topographic details in the assessment of the location and size of the primary tumor and metastases. The adrenomedullary disease is usually multicentric and bilateral, often detected after the onset of MCT; this disease is sought by measurement of urinary metanephrines and fractionated catecholamines. The tumor should be localised by computed tomography or MRI scans; 131I-MIBG scintigraphy is used to confirm diagnosis. Primary hyperpathyroidism generally have no symptoms, although hypercalciuria and renal calculi may occur; we screen for this disease by measurement of serum calcium, once hypercalcemia is documented, serum intact PTH should be measured to confirm the diagnosis. High-resolution small part sonography is sometimes used to differentiate parathyroid hyperplasia from solitary adenoma. PMID- 15111836 TI - [Systemic Salmonella Arizona infection: description of a rare surgical case]. AB - Salmonella arizona enteritis has been described in patients resident in the southern states of the USA and in Mexico, whereas in Europe it is rarer. The virulence of this bacillus is, however, still little known and we have few descriptions of severe systemic infections, which are all present in patients with immune system impairment. Only two cases have been reported in Italy where the infection has occurred as severe sepsis with the pathogenic agent being isolated in the blood. Here we report what is, on the basis of our knowledge, the third case in Italy of a systemic Salmonella arizona infection. PMID- 15111837 TI - [Antithrombotic prophylaxis in day surgery: proposal of a clinical protocol]. AB - The considerable diffusion of the day surgery has new problems such as the use of antithrombotic prophylaxis, now carried out systematically in major surgery. An the basis of the poor data present in the literature, a protocol of study is proposed to evaluate the real effectiveness of this type of prophylaxis in DS, dividing the patients into 3 groups, according to the patient's risk factors to the intervention and to the disease. PMID- 15111838 TI - [Plastic surgery in Milan: an up-date]. PMID- 15111839 TI - Presidential address: Commitment--looking for someone willing to go to the trouble. PMID- 15111840 TI - Iatrogenic operative injuries of abdominal and pelvic veins: a potentially lethal complication. AB - PURPOSE: Epidemiologic studies of vascular injuries are usually limited to those caused by trauma. The purpose of this study was to review the management and clinical outcome in patients with operative injuries to abdominal and pelvic veins. METHODS: Clinical data and outcome in all patients with iatrogenic venous injuries during abdominal and pelvic operations between 1985 and 2002 were reviewed. RESULTS: Forty patients (21 men, 19 women; mean age, 51 years [range, 27-87 years]) sustained 44 venous injuries. Injuries occurred during general (30%), colorectal (23%), orthopedic (20%), gynecologic (15%), and other (12%) operations. Factors leading to injury included oncologic resection (65%), difficult anatomic exposure (63%), previous operation (48%), recurrent tumor (28%), and radiation therapy (20%). All patients had substantial bleeding (mean, 3985 mL; range, 500-20,000 mL). Injuries were located in the inferior vena cava (n = 6), portal vein (n = 7), renal vein (n = 1), and iliac vein (n = 30). Repair was performed with venorrhaphy (64%), end-to-end anastomosis (14%), interposition graft (20%), and vessel ligation (2%). Seven patients (18%) died of injury related causes, including multisystem organ failure (n = 4), uncontrollable bleeding (n = 2), and pulmonary embolism (n = 1). Thirteen patients (32.5%) had major injury-related complications, including repeat exploration because of bleeding (n = 6), multisystem organ failure (n = 6), and venous thrombosis (n = 4). In two patients (5%) unilateral lower extremity edema developed, with no evidence of thrombosis. There was no late graft or venous thrombosis. Variables associated with increased risk for death were massive bleeding, acidosis, hypotension, and hypothermia (P <.05). CONCLUSION: Operative injuries of abdominal and pelvic veins occur in patients undergoing oncologic resection and those with difficult anatomic exposure, owing to previous operation, recurrent tumor, or radiation therapy. Massive blood loss, acidosis, hypotension, and hypothermia are associated with increased risk for death. Repair of venous injuries offers durable results with low incidence of graft or venous thrombosis. PMID- 15111841 TI - Iliac vein compression in an asymptomatic patient population. AB - OBJECTIVE: May-Thurner syndrome is a well-recognized anatomic variant that is associated with the development of symptomatic acute venous thrombosis of the left iliac vein. However, the natural frequency of compression of the left iliac vein and its clinical significance in asymptomatic disease has not been established. Therefore the purpose of this descriptive anatomic study was to determine the incidence of left common iliac vein compression in an asymptomatic population. METHODS: A retrospective analysis of medical records and helical abdominal computed tomography scans was conducted in 50 consecutive patients evaluated in the emergency department because of abdominal pain. Medical records were reviewed for symptoms and risk factors for deep venous thrombosis, and data were collected and reported according to the Joint Society Reporting Standards for acute lower extremity venous thrombosis. All computed tomography was performed with intravenous contrast medium, and 2-mm to 5-mm axial images were obtained. The minor diameter of the common iliac arteries and veins was measured. The technique of transverse image measurement was validated with multiplanar reconstructions and orthogonal diameter measurements in a subset of subjects. Statistical analysis was performed with the Student t test or Spearman rank correlation. RESULTS: Mean age of subjects without symptoms was 40 years (range, 19-85 years), and 60% (n = 30) were female patients. The mean acute lower extremity venous thrombosis risk factor score was 1.16 +/- 0.23 (range, 0-6; maximum possible score, 28). It was surprising that 24% (n = 12) of patients had greater than 50% compression and 66% (n = 33) had greater than 25% compression. Mean compression of the left common iliac vein was 35.5% (range, -5.6%-74.8%). The structure most often compressing the left common iliac vein against the vertebral body was the right common iliac artery (84%). There was no strong correlation between patient age or common iliac artery size and compression of the left common iliac vein. However, women had greater mean compression of the left common iliac vein (women, 41.2% +/- 3.1%; men, 27.0% +/- 3.0%; P =.003). CONCLUSION: Hemodynamically significant left common iliac vein compression is a frequent anatomic variant in asymptomatic individuals. Therefore compression of the left iliac vein may represent a normal anatomic pattern that has thus far been thought of as a pathologic condition. PMID- 15111842 TI - Adrenal insufficiency in patients with ruptured abdominal aortic aneurysms. AB - PURPOSE: Failure of the adrenocortical system after open repair of ruptured abdominal aortic aneurysm (RAAA) has never been reported, to our knowledge. This study was undertaken to examine the incidence and response to treatment of adrenal insufficiency in the RAAA population. METHODS: A 6-year retrospective analysis was carried out on data for all patients admitted after RAAA repair. A cosyntropin stimulation test (CST) was performed in patients with unexplained postoperative hypotension. Patients with adrenal insufficiency were given stress dose hydrocortisone, followed by slow hydrocortisone taper. RESULTS: Twenty of 26 patients admitted after RAAA repair survived longer than 1 week. Nine of these 20 patients underwent CST because of unexplained hypotension, and six patients were found to have adrenal insufficiency. Compared with the three patients with normal CST and the 11 patients with normotension who did not require testing, patients with adrenal insufficiency had greater preoperative hypotension (83% vs 29%; P =.05), greater operative blood loss (7.0 +/- 1.6 L vs 3.0 +/- 0.9 L; P =.003), longer lower extremity ischemia time (5.0 +/- 2.3 hours vs 1.3 +/- 0.5 hours; P =.025), and lower intraoperative urine output (0.8 +/- 0.4 mL/kg/hr vs 2.1 +/- 0.6 mL/kg/hr; P =.023). No difference in length of stay (40 +/- 18 days vs 35 +/- 26 days), major complications (27% vs 32%), or overall mortality (17% vs 15%) was demonstrated with steroid therapy. Initiation of steroid therapy enabled weaning of vasopressor support within 48 hours in patients with adrenal insufficiency. CONCLUSIONS: Adrenal insufficiency was identified in 67% of patients with RAAA with unexplained postoperative hypotension given a CST. Predictors of adrenal insufficiency after RAAA repair include preoperative hypotension and a complicated operative course. Steroid therapy can limit vasopressor dependence, and is not associated with increased morbidity or mortality. PMID- 15111843 TI - Limb salvage after infrainguinal bypass graft failure. AB - OBJECTIVE: The purpose of this study was to examine the outcome of patients in whom an infrainguinal bypass graft failed. METHODS: This was a retrospective analysis of consecutive patients undergoing infrainguinal bypass grafting in a single institution over 8 years. RESULTS: Six hundred thirty-one infrainguinal bypass grafts were placed in 578 limbs in 503 patients during the study period. The indication for surgery was limb-threatening ischemia in 533 patients (85%); nonautologous conduits were used in 259 patients (41%), and 144 (23%) were repeat operations. After a mean follow-up of 28 +/- 1 months (median, 23 months; range, 0-99 months), 167 grafts (26%) had failed secondarily. The rate of limb salvage in patients with graft failure was poor, only 50% +/- 5% at 2 years after failure. The 2-year limb salvage rate depended on the initial indication for bypass grafting: 100% in patients with claudication (n = 16), 55% +/- 8% in patients with rest pain (n = 49), and 34% +/- 6% in patients with tissue loss (n = 73; P <.001). The prospect for limb salvage also depended on the duration that the graft remained patent. Early graft failure (<30 days; n = 25) carried a poor prognosis, with 2-year limb salvage of only 25% +/- 10%; limb salvage was 53% +/- 5% after intermediate graft failure (<2 years, n = 110) and 79% +/- 10% after late failure (>2 years, n = 15; P =.04). Multivariate analysis revealed shorter patency interval before failure (P =.006), use of warfarin sodium (Coumadin) postoperatively (P =.006), and infrapopliteal distal anastomosis (P =.01) as significant predictors for ultimate limb loss. CONCLUSION: The overall prognosis for limb salvage in patients with failed infrainguinal bypass grafts is poor, particularly in patients with grafts placed because of tissue loss and those with early graft failure. PMID- 15111844 TI - Carotid endarterectomy in SAPPHIRE-eligible high-risk patients: implications for selecting patients for carotid angioplasty and stenting. AB - OBJECTIVES: Carotid angioplasty and stenting (CAS) has been proposed as an alternative to carotid endarterectomy (CEA) in patients excluded from the North American Symptomatic Carotid Endarterectomy Trial and the Asymptomatic Carotid Atherosclerosis Study and in those considered at high risk for CEA. In light of recently released CAS data in patients at high risk, we reviewed our experience with CEA. METHODS: The records for consecutive patients who underwent CEA between 1998 and 2002 were retrospectively reviewed, and risk was stratified according to inclusion and exclusion criteria from a "high-risk" or CAS-CEA trial, The Stenting and Angioplasty with Protection in Patients at High Risk for Endarterectomy (SAPPHIRE) trial. RESULTS: Of 776 CEAs performed, 323 (42%) were considered high risk, on the basis of criteria including positive stress test (n = 109, 14%), age older than 80 years (n = 85, 11%), contralateral carotid occlusion (n = 66, 9%), pulmonary dysfunction (n = 56, 7%), high cervical lesion (n = 36, 5%), and repeat carotid operation (n = 27, 3%). Other high-risk criteria included recent myocardial infarction (MI), cardiac surgery, or class III or IV cardiac status; left ventricular ejection fraction less than 30%; contralateral laryngeal palsy; and previous neck irradiation (each <1.5%). Clinical presentation was similar in the high-risk and low-risk groups: asymptomatic (73% versus 73%), transient ischemic attack (23% vs 22%), and previous stroke (4% vs 5%). The overall postoperative stroke rate was 1.4% (symptomatic, 2.9%; asymptomatic, 0.9%). Comparison of high-risk and low-risk CEAs demonstrated no statistical difference in the stroke rate. Factors associated with significantly increased stroke risk included cervical radiation therapy, class III or IV angina, symptomatic presentation, and age 60 years or younger. Overall mortality was 0.3% (symptomatic, 0.5%; asymptomatic, 0.2%), not significantly different between the high-risk (0.6%) and low-risk groups (0.0%). Non-Q-wave MI was more frequent in the high-risk group (3.1 vs 0.9%; P <.05). A composite cluster of adverse clinical events (death, stroke, MI) was more frequent in the symptomatic high-risk group (9.3% vs 1.6%; P <.005), but not in the asymptomatic cohort. There was a trend for more major cranial nerve injuries in patients with local risk factors, such as high carotid bifurcation, repeat operation, and cervical radiation therapy (4.6% vs 1.7%; P <.13). In 121 patients excluded on the basis of synchronous or immediate subsequent operations, who also would have been excluded from SAPPHIRE, the overall rates for stroke (1.65%; P =.69), death (1.65%; P =.09), and MI (0.83%; P =.71) were not significantly different from those in the study population. CONCLUSIONS: CEA can be performed in patients at high risk, with stroke and death rates well within accepted standards. These data question the use of CAS as an alternative to CEA, even in patients at high risk. PMID- 15111846 TI - Reduction in cardiovascular events after vascular surgery with atorvastatin: a randomized trial. AB - OBJECTIVES: This prospective, randomized, placebo-controlled, double-blind clinical trial was performed to analyze the effect of atorvastatin compared with placebo on the occurrence of a 6-month composite of cardiovascular events after vascular surgery. Cardiovascular complications are the most important cause of perioperative morbidity and mortality among patients undergoing vascular surgery. Statin therapy may reduce perioperative cardiac events through stabilization of coronary plaques. METHODS: One hundred patients were randomly assigned to receive 20 mg atorvastatin or placebo once a day for 45 days, irrespective of their serum cholesterol concentration. Vascular surgery was performed on average 30 days after randomization, and patients were prospectively followed up over 6 months. The cardiovascular events studied were death from cardiac cause, nonfatal myocardial infarction, unstable angina, and stroke. RESULTS: Fifty patients received atorvastatin, and 50 received placebo. During the 6-month follow-up primary end points occurred in 17 patients, 4 in the atorvastatin group and 13 in the placebo group. The incidence of cardiac events was more than three times higher with placebo (26.0%) compared with atorvastatin (8.0%; P =.031). The risk for an event was compared between the groups with the Kaplan-Meier method, as event-free survival after vascular surgery. Patients given atorvastatin exhibited a significant decrease in the rate of cardiac events, compared with the placebo group, within 6 months after vascular surgery (P =.018). CONCLUSION: Short-term treatment with atorvastatin significantly reduces the incidence of major adverse cardiovascular events after vascular surgery. PMID- 15111848 TI - Can low molecular weight heparin replace unfractionated heparin during peripheral arterial reconstruction? An open label prospective randomized controlled trial. AB - OBJECTIVE: This study was undertaken to evaluate the effect of low molecular weight heparin (LMWH) compared with unfractionated heparin (UFH) on the rate of occlusion and bleeding during peripheral vascular surgery. METHODS: The study was an open label, prospective, randomized trial, carried out by 20 Swedish surgical and vascular surgical departments that report to the Swedish Vascular Registry (SWEDVASC). Study subjects included patients undergoing peripheral vascular procedures, except carotid surgery. Of the 849 patients included, 817 were followed up to 30 days. LMWH (40 mg of enoxaparin) or UFH (5000 IU heparin) was given intravenously immediately before clamping. The same formulation in diluted form was used for vascular rinsing. Main outcome measures included patent reconstruction at day 1, perioperative blood loss, and the percentage of patients requiring protamin. Further, 30-day data for mortality, repeat operation, and recurrent occlusion are reported. RESULTS: The mortality rate at 30 days was 2.7%, with no difference between groups. The patency rate at 1 day was 91.2% to 98.4%, depending on diagnosis and type of reconstruction. No difference was recorded between study groups (0.6 < P < 1.0). At 30 days the patency rate was 83.1% to 100% (0.2 < P <.9). Median blood loss was 350 mL (interquartile range [IQR], 200-800 mL) in the LMWH group and 425 mL (IQR, 200-900 mL) in the UFH group (P =.02). Protamin was given to significantly fewer patients in the LMWH group (P =.001). LMWH was comparable to UFH during peripheral vascular reconstruction in terms of 1-day and 30-day graft patency, operative blood loss, and hemorrhagic complications. Protamine was required less often after LMWH. In this randomized trial LMWH was as effective as UFH in preventing thrombosis without excess bleeding or hemorrhagic complications. PMID- 15111849 TI - Randomized trial of vein versus dacron patching during carotid endarterectomy: long-term results. AB - BACKGROUND AND PURPOSE: Overviews of randomized patch trials by the Cochrane Collaboration suggest that a policy of routine patching is preferable to routine primary closure. However, there is no systematic evidence that patch type, whether prosthetic or vein, influences outcome after carotid endarterectomy (CEA). METHODS: Two hundred seventy-three patients were randomized to vein or thin-walled Dacron patch (Hemashield Finesse) closure of the arteriotomy after 276 CEA procedures. Patients were reviewed clinically and with duplex ultrasound scanning at 1, 6, 12, 24, and 36 months or until death. No patients were lost to follow-up. Cumulative statistical analyses are presented for the 264 patients (269 CEAs) who actually received a randomized treatment allocation. RESULTS: Cumulative freedom from death or ipsilateral stroke at 3 years (including operative events) was 93.0% in the Dacron patch group and 95.5% in the vein group P =.42). Cumulative freedom from death or any stroke was 91.5% after Dacron patch closure and 93.9% after vein closure (P =.46). Cumulative freedom from recurrent stenosis greater than 70% or occlusion at 3 years was 92.9% for patients randomized to the Dacron patch group and 98.4% for patients randomized to the vein group (P =.03). At 3 years the incidence of stroke in the carotid territory not operated on was 1.0% in 93 patients with no contralateral internal carotid artery disease at randomization, and increased to 1.3% in 78 patients with 1% to 69% stenosis, and 2.0% in 51 patients with contralateral 70% to 99% stenosis. No late strokes occurred distal to 42 occluded contralateral internal carotid arteries. CONCLUSIONS: Patch type has no influence on early operative risk, no association with enhanced patterns of thrombogenicity in the early postoperative period, and no influence on risk for ipsilateral or any stroke at 3 years. Dacron patches were, however, associated with a significantly higher incidence of recurrent stenosis at 3 years, with most occurring within 6 to 12 months of surgery. However, the higher incidence of recurrent stenosis was not associated with a parallel increase in late stroke, and in this study a program of serial ultrasound surveillance could not have prevented one ipsilateral stroke. PMID- 15111851 TI - A new approach to carotid angioplasty and stenting with transcervical occlusion and protective shunting: Why it may be a better carotid artery intervention. AB - OBJECTIVE: The purpose of this study was to evaluate the effectiveness and demonstrate the advantages of a new technique for carotid angioplasty and stenting (CAS) with proximal cerebral protection through a direct transcervical approach, as compared with a percutaneous transfemoral approach. METHODS: CAS procedures were carried out in 25 consecutive patients, 4 with the femoral approach and 21 through a 2-cm incision at the base of the neck, with the patient under local anesthesia. For transcervical occlusion and protective shunting (TOPS), a short 9F sheath was inserted directly into the common carotid artery and connected to a 6F sheath placed percutaneously in the ipsilateral internal jugular vein. After clamping the common carotid artery proximal to the 9F sheath, internal carotid artery blood flow reversal was confirmed or an occluding external carotid balloon was placed. A filter interposed between the arterial and venous sheaths collected embolic debris from transcarotid manipulations. The arterial puncture was directly repaired with suture. Neurologic status was assessed with the National Institutes of Health stroke scale by an independent neurology consultant before and after the procedure. RESULTS: One of the four percutaneous femoral approaches that failed because of tortuous anatomy was successfully treated with TOPS. Angiographic confirmation demonstrating resolution of asymptomatic (>80%; n = 12) stenosis or symptomatic (>60%; n = 12) stenosis was achieved in all patients with stents. A 0% technical failure rate and 0% combined 30-day stroke or mortality rate were achieved in all CAS attempted with TOPS. There were no hematomas in the cervical group, despite pretreatment with clopidogrel bisulfate and heparin, and one hematoma in the femoral group after failure of a Perclose arterial closure device. In one of the patients in the femoral group bilateral cholesterol emboli to the toes developed. CONCLUSION: TOPS solves problems of access, embolization into the cerebral and peripheral circulation, and specialized cerebral protection devices, and enables secure closure of the access vessel in patients given anticoagulation therapy. TOPS may provide a safer, more effective, economical means for performing CAS. PMID- 15111852 TI - Octogenarians with contralateral carotid artery occlusion: a cohort at higher risk for carotid endarterectomy? AB - PURPOSE: Carotid angioplasty and stenting has been proposed as a treatment option for carotid occlusive disease in patients at high risk, including those 80 years of age or older or with contralateral carotid occlusion. We analyzed 30-day mortality and stroke risk rates of carotid endarterectomy (CEA) in patients aged 80 years or older with concurrent carotid occlusive disease. METHODS: From a retrospective review of 1000 patients undergoing 1150 CEA procedures to treat symptomatic and asymptomatic carotid lesions over 13 years, we identified 54 patients (5.4%) aged 80 years or older with concurrent contralateral carotid occlusion. These patients were compared with 38 patients (3.8%) aged 80 years or older with normal or diseased patent contralateral carotid artery and 81 patients (8.1%) younger than 80 years with contralateral carotid occlusion. All CEA procedures involved either standard CEA with patching or eversion CEA, and were performed by the same surgeon, with the patients under deep general anesthesia and cerebral protection involving continuous perioperative electroencephalographic monitoring for selective shunting. Shunting criteria were based exclusively on electroencephalographic abnormalities consistent with cerebral ischemia. RESULTS: The 30-day mortality and stroke rate in patients aged 80 years or older with concurrent contralateral carotid occlusion was zero. CONCLUSIONS: The concept of high-risk CEA needs to be revisited. Patients with two of the criteria considered high risk in the medical literature, that is, age 80 years or older and contralateral carotid occlusion, can undergo CEA with no greater risks or complications. Until prospective randomized trials designed to evaluate the role of carotid angioplasty and stenting have been completed, CEA should remain the standard treatment in such patients. PMID- 15111853 TI - Allograft replacement for infrarenal aortic graft infection: early and late results in 179 patients. AB - OBJECTIVES: We evaluated early and late results of allograft replacement to treat infrarenal aortic graft infection in a large number of patients and compared the results in patients who received fresh allografts versus patients who received cryopreserved allografts. METHODS: From 1988 to 2002 we operated on 179 consecutive patients (mean age, 64.6 +/- 9.0 years; 88.8% men). One hundred twenty-five patients (69.8%) had primary graft infections, and 54 patients (30.2%) had secondary aortoenteric fistulas (AEFs). Fresh allografts were used in 111 patients (62.0%) until 1996, and cryopreserved allografts were used in 68 patients (38.0%) thereafter. RESULTS: Early postoperative mortality was 20.1% (36 patients), including four (2.2%) allograft-related deaths from rupture of the allograft (recurrent AEF, n = 3), all in patients with fresh allografts. Thirty two deaths were not allograft related. Significant risk factors for early mortality were septic shock (P <.001), presence of AEF (P =.04), emergency operation (P =.003), emergency allograft replacement (P =.0075), surgical complication (P =.003) or medical complication (P <.0001), and need for repeat operation (P =.04). There were five (2.8%) nonlethal allograft complications (rupture, n = 2; thromboses, which were successfully treated at repeat operation, n = 2; and amputation, n = 1), all in patients with fresh allografts. Four patients (2.2%) were lost to follow-up. Mean follow-up was 46.0 +/- 42.1 months (range, 1-148 months). Late mortality was 25.9% (37 patients). There were three (2.1%) allograft-related late deaths from rupture of the allograft, at 9, 10, and 27 months, respectively, all in patients with fresh allografts. Actuarial survival was 73.2% +/- 6.8% at 1 year, 55.0% +/- 8.8% at 5 years, and 49.4% +/- 9.6% at 7 years. Late nonlethal aortic events occurred in 10 patients (7.2%; occlusion, n = 4; dilatation < 4 cm, n = 5; aneurysm, n = 1), at a mean of 28.3 +/- 28.2 months, all but two in patients with fresh allografts. The only significant risk factor for late aortic events was use of an allograft obtained from the descending thoracic aorta (P =.03). Actuarial freedom from late aortic events was 96.6% +/- 3.4% at 1 year, 89.3% +/- 6.6% at 3 years, and 89.3% +/- 6.6% at 5 years. There were 63 late, mostly occlusive, iliofemoral events, which occurred at a mean of 34.9 +/- 33.7 months in 38 patients (26.6%), 28 of whom (73.7%) had received fresh allografts. The only significant risk factor for late iliofemoral events was use of fresh allografts versus cryopreserved allografts (P =.03). Actuarial freedom from late iliofemoral events was 84.6% +/- 7.0% at 1 year, 72.5% +/- 9.0% at 3 years, and 66.4% +/- 10.2% at 5 years. CONCLUSIONS: Early and long-term results of allograft replacement are at least similar to those of other methods to manage infrarenal aortic graft infections. Rare specific complications include early or late allograft rupture and late aortic dilatation. The more frequent late iliofemoral complications may be easily managed through the groin. These complications are significantly reduced by using cryopreserved allografts rather than fresh allografts and by not using allografts obtained from the descending thoracic aorta. PMID- 15111854 TI - National Audit of Thrombolysis for Acute Leg Ischemia (NATALI): clinical factors associated with early outcome. AB - OBJECTIVE: The National Audit of Thrombolysis for Acute Leg Ischemia (NATALI) database is a consecutive series of patients who underwent intra-arterial thrombolysis to treat acute leg ischemia in one of 11 centers in the United Kingdom. The purpose of the study was to analyze the factors associated with outcome after 30 days. METHODS: The data were collected over 10 years on standard pro formas, and registration was completed at the end of 1999. Since then, data from each unit have been verified and missing data included when available. Univariate and multivariate analyses were performed, with the outcomes of amputation-free survival (AFS), amputation with survival, and death. RESULTS: A total of 1133 thrombolytic events were included. Outcome results at 30 days for the entire group were AFS, 852 (75.2%); amputation, 141 (12.4%); and death, 140 (12.4%). Results for the entire group improved from the first half of the database, when AFS ranged from 65% to 75%, to almost 80% for the last few years of the study, although this was not statistically significant. Preintervention factors associated with lower AFS at multivariate analysis included diabetes (P =.002), increasing age (P <.001), short-duration ischemia (P =.027), Fontaine grade (P =.001), and ischemia with neurosensory deficit (P =.004). AFS was improved in patients receiving warfarin sodium at the time of the arterial occlusion (P =.04). Mortality was higher in women (P =.006) and in older patients (P <.001), and in patients with native vessel occlusion (P <.001), emboli (P =.02), or a history of ischemic heart disease (P <.001). Amputation risk was greatest in younger men (P <.001) and in patients with more severe ischemia (P =.02), graft occlusion (P <.001), or native vessel thrombotic occlusion (P =.02). CONCLUSION: Experienced surgeons and radiologists can achieve an AFS of about 80% in selected patients with acute leg ischemia. Information from the NATALI database can be used in selection of an appropriate intervention in the individual patient. PMID- 15111856 TI - Bypass versus angioplasty to treat severe limb ischemia: factors that affect treatment preferences of UK surgeons and interventional radiologists. AB - OBJECTIVE: There is continuing controversy as to whether surgical bypass or angioplasty should be first-line treatment of severe limb ischemia. We undertook this study to examine angiographic and clinical factors that influence the treatment of severe limb ischemia by vascular surgeons and interventional radiologists. METHODS: Twenty consultant vascular surgeons and 17 consultant vascular interventional radiologists evaluated 596 hypothetical clinical or angiographic scenarios, and recorded whether, in their opinion, the most appropriate first-line treatment was surgical bypass, angioplasty, or primary amputation. Stepwise multiple linear regression was used to identify the factors that significantly affected responses from the entire group and from surgeons and radiologists separately. RESULTS: There were significant differences between surgeons and radiologists with regard to how clinical and angiographic variables determined treatment preferences. Increasing disease severity, absence of runoff into the foot, presence of a suitable vein, and tissue loss as opposed to rest pain only (the latter only significant to surgeons) all increased the response score toward surgery. However, surgeons and radiologists weighted each of these factors quite differently. Even in the most complex statistical model, 19% of surgical and 13% of radiologic response variations remained unexplained. CONCLUSIONS: Individual surgeons and radiologists vary considerably in their views of the relative merits of surgery and angioplasty in patients with severe limb ischemia. This broad gray area mandates the need for randomized controlled trial data to inform joint decision-making and to optimize patient outcome. PMID- 15111857 TI - Changes in blood coagulability as it traverses the ischemic limb. AB - OBJECTIVE: We undertook this study to determine whether changes in blood coagulability associated with peripheral arterial occlusive disease are due to contact with the atherosclerotic arterial wall or passage through distal ischemic tissue. METHODS: Thirty patients with peripheral arterial occlusive disease undergoing angiography participated in the study. Ankle-brachial pressure index was recorded before intervention. Blood samples taken from the aorta, common femoral artery, and common femoral vein were analyzed at thromboelastography. Angiograms were scored for stenotic disease by a radiologist blinded to the other results. RESULTS: When femoral artery samples were compared with aortic samples there was a decrease in reaction time (R; P <.05), an increase in maximum amplitude (MA; P <.05), and an increase in coagulation index (CI; P <.002), indicating an increase in coagulability as blood flowed down the iliac segment. These changes also correlated (DeltaR, r = 0.442, P <.05; DeltaMA, r = 0.379, P <.05; DeltaCI, r = 0.429, P <.05) with the severity of disease in the ipsilateral iliac segment. Significant differences in R (P <.05), angle (P <.05), MA (P <.005), and CI (P <.001) between common femoral arterial and venous samples confirmed that venous samples were more coagulable in this group of patients. This difference in Thromboelastography parameters across the arteriovenous segment correlated inversely with the degree of ischemia (represented by ankle brachial pressure index; DeltaCI, r = -0.427, P <.05; DeltaMA, r = -0.370, P <.05) in the puncture side limb. CONCLUSION: Passage of blood down an atherosclerotic artery leads to an increase in coagulability proportional to the degree of stenosis in that vessel. Passage of blood through ischemic tissue may also contribute to increased coagulability in peripheral arterial occlusive disease. PMID- 15111858 TI - Basilic vein transposition fistula: a good option for maintaining hemodialysis access site options? AB - PURPOSE: The primary use of autogenous arteriovenous access for chronic hemodialysis is recommended by the National Kidney Foundation-Dialysis Outcomes Quality Initiative practice guidelines. We review the outcomes of basilic vein transposition (BVT) to assess its value as a primary upper arm arteriovenous access option. METHODS: A retrospective review of 56 patients undergoing BVT was performed. Thirty patients were men; average age was 56 years. Etiology of end stage renal disease, complications, and time to maturation were tabulated. Primary and secondary patency rates were determined by using life table methods. Multivariate regression analysis was performed to assess risk factors for fistula failure. RESULTS: Renal failure was associated with diabetes in 32 (57%) patients, and BVT was the primary access procedure in 22 (39%) patients. Perioperative complications occurred in 5 (9%) patients and included hematoma (n = 3), myocardial infarction (n = 1), and death (n = 1). The average time to maturation was 74 days (range, 12-265 days), and maturation failure occurred in 21 (38%) patients. Logistic regression analysis showed that age older than 60 years was associated with poorer maturation and patency rates. On an intent-to treat basis, 1-year primary and secondary patencies were 35% and 47%, respectively, but only 18% and 28%, respectively, for age >60 years. Forty-two percent of failed BVT were subsequently replaced with a prosthetic graft by using the same upper arm vessels. CONCLUSION: BVT frequently do not mature in patients older than 60 years, which compromises its utility as a primary access. However, fistulas that mature provide acceptable patency rates, and subsequent conversion to a prosthetic access is frequently possible. Selective use of BVT might improve the utilization of available access sites. PMID- 15111859 TI - Surgical disobliteration of postthrombotic deep veins--endophlebectomy--is feasible. AB - OBJECTIVE: Partial obstruction of postthrombotic veins is caused by endovenous scar tissue, which creates synechiae and septae that narrow and sometimes block the lumen. We have performed surgical disobliteration, or endophlebectomy, of chronically obstructed venous segments during various kinds of deep venous reconstructions to increase the flow through previously obstructed segments. In this article we describe the endophlebectomy technique, and report the availability of this procedure as an adjunct to deep venous reconstructions for the treatment of postthrombotic chronic venous insufficiency. Patients and methods Between July 1996 and February 2003, surgical disobstruction of 23 deep venous segments was performed in 13 patients in association with 14 deep venous reconstructions to treat advanced postthrombotic chronic venous insufficiency. Postthrombotic veins were surgically exposed, and a longitudinal venotomy was carried out at a variable length. The synechiae and masses attached to the intimal layer were carefully excised. Mean duplex scanning follow-up was 10.8 +/- 8.2 months (median, 8 months; range, 1-28 months). RESULTS: In 10 patients (77%) the treated segments remained primarily patent at median follow-up of 8 months (range, 1-28 months). Early thrombosis near the endophlebectomy site occurred in 3 patients, at 2, 5, and 12 days, respectively, after surgery. In 2 patients with early thrombosis further interventions were carried out with success. In a third patient with early postoperative thrombosis the final outcome was recanalization and reflux. These results yielded an overall secondary patency rate of 93%. No perioperative pulmonary embolism was observed. CONCLUSION: This series demonstrates that surgical disobliteration of postthrombotic deep veins is technically feasible, and led to patency of the segments for the duration of follow-up for up to 28 months (mean, 10.8 +/- 8.2 months). We used this technique with the objective of disobstructing postthrombotic veins, to increase flow through a previously narrowed lumen. Postoperative thrombosis at the site of endophlebectomy occurred in 23% of patients. Although this early experience is encouraging, further studies and longer follow-up are necessary to assess the durability of the procedure. PMID- 15111861 TI - Correlation of duplex ultrasound scanning-derived valve closure time and clinical classification in patients with small saphenous vein reflux: Is lesser saphenous vein truly lesser? AB - OBJECTIVE: We recently identified small saphenous vein (SSV) reflux as a significant risk factor for ulcer recurrence in patients with severe chronic venous insufficiency (CVI) undergoing perforator vein ligation. In this study we examined the role of SSV reflux in patients across the spectrum of CVI. METHODS: From March 15, 1997, to December 24, 2002, clinical and duplex ultrasound (US) scanning data from all valve closure time studies performed in our vascular laboratory were prospectively recorded. Valve closure time in the deep and superficial leg veins was assessed with the rapid cuff deflation technique; reflux time greater than 0.5 seconds was considered abnormal. SSV reflux was correlated with the CEAP classification system and eventual surgical procedure. Data were analyzed with Pearson chi(2) analysis. RESULTS: We analyzed 722 limbs in 422 patients, 265 (63%) female patients and 157 (37%) male patients, with a mean age of 48 +/- 12.8 years (range, 16-85 years). In the entire cohort the cause was congenital (Ec) in 5 patients, primary (Ep) in 606 patients, and secondary (Es) in 112 patients. SSV reflux was present in 206 limbs (28.5%) evaluated. Among limbs with SSV reflux, Ec = 4 (2%), Ep = 162 (79%), and Es = 40 (19%). SSV reflux did not correlate with gender, side, or age. The prevalence of SSV reflux increases with increasing severity of clinical class: C1-C3, 25.8% versus C4-C6, 36.1% (P =.006). SSV reflux is highly associated with deep venous reflux, 35.2% of femoral vein reflux (P =.015), 35.8% of femoral vein plus popliteal vein reflux (P =.001), and 40.5% of isolated popliteal vein reflux (P <.001). Great saphenous vein (GSV) reflux was identified in 483 (67%) limbs studied with valve closure time, whereas SSV reflux was present in 206 (28%) limbs. In this cohort, 127 GSV or SSV surgical procedures were performed subsequent to valve closure time examination. Among these operations 107 (84%) were GSV procedures, and only 20 (16%) were SSV procedures. CONCLUSION: SSV reflux is most common in patients demonstrating severe sequelae of CVI, such as lipodermatosclerosis or ulceration. The increasing prevalence of SSV reflux in more severe clinical classes and the strong association of SSV reflux and deep venous reflux suggest that SSV may have a significant role in CVI. Our data further show that, in our institution, a GSV with reflux is more than twice as likely to be surgically corrected as an SSV with reflux. It is time for the SSV to assume greater importance in the treatment of lower extremity venous disease. Future improvements in surgical techniques for access and visualization of the SSV may facilitate this method. PMID- 15111862 TI - Silyl-heparin bonding improves the patency and in vivo thromboresistance of carbon-coated polytetrafluoroethylene vascular grafts. AB - OBJECTIVES: Our purpose was to improve the performance of carbon-coated expanded polytetrafluoroethylene vascular grafts by bonding the grafts with silyl-heparin, a biologically active heparin analog, using polyethylene glycol as a cross linking agent. Material and method Silyl-heparin-bonded carbon-coated expanded polytetrafluoroethylene vascular grafts (Bard Peripheral Vascular, Tempe, Ariz), were evaluated for patency and platelet deposition 2 hours, 7 days, and 30 days after graft implantation in a canine bilateral aortoiliac artery model. Platelet deposition was determined by injection of autologous, (111)Indium-radiolabeled platelets, followed by a 2-hour circulation period prior to graft explantation. Histologic studies were performed on a 2-mm longitudinal strip of each graft (7 day and 30-day groups). Heparin activity of the explanted silyl-heparin grafts was determined by using an antithrombin-III based thrombin binding assay. RESULTS: Overall chronic graft patency (7-day and 30-day groups) was 100% for the silyl-heparin bonded (16/16) grafts versus 68.75% for control (11/16) grafts (P =.043). Acute 2-hour graft patency was 100% for the silyl-heparin bonded (6/6) grafts versus 83.3% for control (5/6) grafts. Radiolabeled platelet deposition studies revealed a significantly lower amount of platelets deposited on the silyl heparin grafts as compared with control grafts in the 30-day group (13.8 +/- 7.18 vs 28.4 +/- 9.73, CPM per cm2 per million platelets, mean +/- SD, P =.0451, Wilcoxon rank sum test). In the 2-hour group of dogs, a trend towards a lower deposition of platelets on the silyl-heparin grafts was observed. There was no significant difference in platelet deposition between the two grafts in the 7-day group. Histologic studies revealed a significant reduction in intraluminal graft thrombus present on the silyl-heparin grafts as compared with control grafts in the 30-day group of animals. In contrast, there was no difference in amount of graft thrombus present on both graft types in the 7-day group of dogs. Pre implant heparin activity on the silyl-heparin bonded grafts was 2.0 IU/cm(2) (international units[IU]/cm(2)). Heparin activity remained present on the silyl heparin grafts after explantation at all 3 time points (2 hours: above upper limit of assay, upper limit = 0.57, n = 6; 7 days: 0.106 +/- 0.015, n = 5; 30 days: 0.007 +/- 0.001, n = 5; mean +/- SD, IU/cm(2)). CONCLUSION: Silyl-heparin bonding onto carbon-coated expanded polytetrafluoroethylene vascular grafts resulted in (1) improved graft patency, (2) increased in vivo graft thromboresistance, and (3) a significant reduction in intraluminal graft thrombus. This graft may prove to be useful in the clinical setting. PMID- 15111863 TI - Endograft technology: a delivery vehicle for intravascular gene therapy. AB - PURPOSE: The purpose of this study was to determine whether vascular smooth muscle cells (SMCs) suffused into a bilayered stent graft retain and express a retrovirally transduced gene for 7 months in vivo. METHODS: SMCs harvested from dog jugular vein were retrovirally transduced to introduce genes for tissue plasminogen activator (t-PA) and beta-galactosidase. These cells were then suffused into a novel dual-layered Dacron graft and cultured for 36 to 48 hours. The grafts were mounted on a Palmaz stent and balloon- expanded in the infrarenal aorta of the SMC donor dogs (n = 6). Grafts were recovered at 1, 2, 3, 4, 5, and 7 months. A control endograft suffused with SMCs transduced with only the beta galactosidase gene was placed in the dogs with grafts recovered at 2, 3, and 4 months. t-PA antigen concentration and expression were analyzed with an enzyme linked immunosorbent assay. RESULTS: Retained engineered SMCs (blue nuclei) were identified in the explanted grafts, neointima, and underlying aorta with X-gal staining. The t-PA antigen concentration and t-PA activity from the SMCs recovered from the grafts remained elevated for the duration of the experiment (7 months) at levels significantly higher (3.7 +/- 0.2 ng/mL per 10(5) cells per 24 hours and 1.4 +/- 0.1 IU/mL per 10(5) cells per 24 hours) than in control endografts (0.5 +/- 0.03 ng/mL per 10(5) cells per 24 hours and 0.07 +/- 0.00 IU/mL per 10(5) cells per 24 hours; P <.001). No graft stenosis was observed. CONCLUSION: Retrovirally engineered vascular SMCs survived the implantation trauma, repopulated each graft, migrated into the underlying aorta, and expressed the transduced genes for the 7-month duration of the experiment. This bilayered Dacron endograft model provides a platform to study direct intravascular gene therapy. PMID- 15111864 TI - Recombinant human thrombomodulin inhibits arterial neointimal hyperplasia after balloon injury. AB - OBJECTIVE: Smooth muscle cell proliferation is a major pathophysiologic factor in injury-induced neointimal hyperplasia and recurrent stenosis. We have demonstrated that recombinant human thrombomodulin (rTM) inhibits thrombin induced arterial smooth muscle cell proliferation in vitro. The purpose of this study was to investigate the effect of rTM on neointimal hyperplasia in vivo. METHODS: A rabbit femoral artery balloon injury model was used. Bilateral superficial femoral arteries were deendothelialized with a 2F arterial embolectomy catheter. rTM (145 microg/kg; 2.0 microg/mL in circulation) or Tris hydrochloride vehicle control was administered intravenously during the procedure, then either discontinued (group A) or administered twice daily for an additional 48 hours (group B). Rabbits were euthanized at 4 days and at 1, 2, and 4 weeks, and femoral artery specimens were prepared with in situ perfusion fixation and paraffin embedding. Luminal, intima, media, and whole artery areas were quantitated with digital imaging computerized planimetry. Intima-media and lumen-whole artery ratios were calculated. The injury-induced inflammatory reaction was also evaluated with light microscopy, scanning and transmission electron microscopy, and immunohistochemical and immunohistofluorescence staining. RESULTS: In the buffer control group, neointimal hyperplasia after femoral artery balloon injury was evident at 2 weeks, and was pronounced at 4 weeks (P <.0001). Infusion of rTM significantly inhibited intimal hyperplasia at both 2 and 4 weeks (P <.0001). In group A, rTM reduced the intima-media ratio by 27% and 39% at 2 and 4 weeks, respectively. Extended administration of rTM (group B) resulted in inhibition of hyperplasia by 57% and 30% at 2 and 4 weeks, respectively, but failed to reach significance compared with the shorter exposure. rTM infusion significantly inhibited thrombosis (8.1-fold) compared with the buffer control group (P =.012). rTM had no significant effect on lumen area or lumen-whole artery ratio, but treated arteries demonstrated significantly less compensatory dilatation (P =.045), as measured by whole artery area in response to less intimal hyperplasia. rTM administration inhibited platelet adhesion and inhibition of neutrophil infiltration to a degree that approached statistical significance (P =.0675). CONCLUSIONS: Systemic intravenous administration of rTM significantly decreases neointimal hyperplasia and improves patency in the rabbit femoral artery after balloon injury. In addition to exhibiting antithrombotic and antiproliferative effects, rTM may also invoke an anti-inflammatory mechanism, and may alter vascular remodeling in a multidimensional role to inhibit recurrent stenosis after arterial injury. PMID- 15111865 TI - Differential expression and activity of matrix metalloproteinases during flow modulated vein graft remodeling. AB - OBJECTIVE: While shear stress closely regulates vascular remodeling, the mediators of this process have been only partially elucidated. The current study examined the role of the gelatinases in flow-mediated vein graft intimal hyperplasia. We hypothesized that matrix metalloproteinase (MMP)-2 and MMP-9 expression and protein levels, relative to tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2, are upregulated in a flow-dependent manner during vein graft arterialization. METHODS: Bilateral common carotid interposition vein grafting was performed in rabbits. Reduction in flow was achieved through unilateral ligation of the internal carotid artery and three of four branches of the external carotid artery. At 28 days grafts were harvested and analyzed for intimal area; MMP-2 and MMP-9, and TIMP-1 and TIMP-2 messenger RNA content, via quantitative reverse transcription polymerase chain reaction; and MMP-2 and MMP 9, and TIMP-1 and TIMP-2 protein concentrations, via both bioactivity assay and zymography. RESULTS: Branch ligation resulted in a 10-fold difference in mean flow rate and accelerated development of intimal hyperplasia in a low-flow environment. Exposure of the vein graft to arterial hemodynamics induced a marked rise in MMP-9 mRNA levels, whereas only a modest increase in MMP-2 mRNA was observed. MMP-2 protein was 50 to 100 times more abundant than MMP-9, and was significantly upregulated in grafts that demonstrated enhanced intimal thickening. Immunohistochemistry demonstrated that MMP-2 was located throughout the myointima, whereas MMP-9 was localized almost exclusively to the region of endothelium. No differences in TIMP-1 and TIMP-2 mRNA or protein levels were detected between high-flow and low-flow grafts. CONCLUSION: MMP-2 is the predominate gelatinase that regulates early vein graft remodeling. Despite a marked increase in MMP-9 gene expression, development of intimal hyperplasia after a reduction in wall shear rate correlates with an increase in MMP-2 protein levels. These data suggest differential regulatory mechanisms for proteases within the remodeling vein graft wall. Modulation of extracellular matrix biologic features may offer therapeutic strategies for the prevention of vein graft failure. PMID- 15111866 TI - Interleukin-1beta inhibits PDGF-BB-induced migration by cooperating with PDGF-BB to induce cyclooxygenase-2 expression in baboon aortic smooth muscle cells. AB - OBJECTIVE: Smooth muscle cell (SMC) migration from the media into the intima is pivotal for intimal formation after vascular injury. Platelet-derived growth factor (PDGF)-BB is a potent chemoattractant for SMCs in vitro and in vivo. We investigated whether interleukin (IL)-1beta affects migration in response to PDGF BB. Our data suggest that IL-1beta is inhibitory and that this effect is mediated by cyclooxygenase (COX)-2. We further addressed the role of the mitogen-activated protein kinase p38, which is activated by PDGF-BB and by IL-1beta. METHODS: Baboon aortic SMCs were prepared with the explant method. Migration was measured in a Boyden chamber assay through filters coated with monomeric collagen. COX2 expression and phosphorylation of p38 MAPK were analyzed by Western blotting. RESULTS: PDGF-BB (10 ng/mL) stimulates migration 3.8-fold, and IL-1beta (0.1 ng/mL) reduces this response by 40%. The inhibitory effect of IL-1beta is abolished by the COX inhibitor, indomethacin (10 micromol/L), the specific COX2 inhibitor, NS398 (10 micromol/L), and the p38 MAPK inhibitor SB203580 (3 micromol/L). We found that IL-1beta and PDGF-BB synergize to stimulate COX2 expression. We further demonstrated that p38 MAPK is activated by IL-1beta and PDGF with different kinetics and that p38 MAPK is required for maximal COX2 expression in response to IL-1beta plus PDGF-BB. CONCLUSION: IL-1beta inhibits PDGF-BB-induced migration by cooperating with PDGF-BB to induce COX2 through activation of p38 MAPK. Whether this effect of IL-1beta modulates intimal growth after vascular injury remains to be elucidated. PMID- 15111867 TI - Gamma-irradiation modulates vascular smooth muscle cell and extracellular matrix function: Implications for neointimal development. AB - OBJECTIVE: Migration of vascular smooth muscle cells (SMCs) into the subintimal space, and their proliferation and resultant deposition of extracellular matrix are key processes in the development of intimal hyperplasia, leading to vascular recurrent stenosis. The purpose of this study was to investigate the effects of clinically administered doses of gamma-radiation on SMCs and extracellular matrix proteins in vitro, to better understand how it impinges on cellular and extracellular components of recurrent stenosis. METHODS: The effects of gamma irradiation (10, 20 Gy) on SMC migration into three-dimensional collagen matrix gels was quantitated by calibrated light microscopy, and the release of metalloproteinases into conditioned media was investigated with an enzyme-linked immunosorbent assay and zymography. Collagen production was assayed with [(3)H] proline incorporation, and SMC phenotype changes with confocal microscopy with a fluorescent alpha-actin antibody. The effect of gamma-irradiation on extracellular matrix was investigated by quantitating untreated SMC proliferation ((3)H-thymidine incorporation) on irradiated endothelial cell-derived matrix and by assessing structural collagen matrix changes with sodium dodecylsulfate polyacrylamide gel electrophoresis. All groups were compared with nonirradiated control groups. RESULTS: SMC vertical migration was significantly decreased by gamma-irradiation (48% and 55%, respectively; P <.0001). Irradiation did not generate measurable matrix protein crosslinks, nor did it alter the production of metalloproteinases or collagen synthesis. However, gamma-irradiation decreased the ability of extracellular matrix to induce nonirradiated SMC proliferation (15% reduction; P =.0028). Moreover, gamma-irradiation reversed the secretory phenotype of cultured SMCs to a contractile type. CONCLUSIONS: The gamma irradiation-induced reduction of cellular migration, changes in SMC phenotype, and functional activity of matrix-bound factors, and no measurable effects on the production of extracellular matrix proteins, may in part explain the diverse effects of gamma-irradiation on the restenotic response. PMID- 15111868 TI - A monoclonal antibody against cytokine-induced neutrophil chemoattractant attenuates injury in the small intestine in a model of ruptured abdominal aortic aneurysm. AB - PURPOSE: Ruptured abdominal aortic aneurysm (RAAA) continues to be a major source of aneurysm-related morbidity and mortality. Neutrophils have been implicated in RAAA repair-induced organ injury; however, the agents responsible for neutrophil activation and organ sequestration have not been identified. This study investigated the role of cytokine-induced neutrophil chemoattractant (CINC) in organ injury in an RAAA model. METHODS: Rats were subjected to 1 hour of hemorrhagic shock with resuscitation, followed by 45 minutes of lower torso ischemia and 2 hours of reperfusion, and randomly were selected to receive saline solution or anti-rat CINC monoclonal antibody at the start of hemorrhagic shock. Another group of animals underwent sham operation, and served as a control group. Intestinal and lung permeability, intestinal and lung myeloperoxidase (MPO) activity, intestinal and lung CINC, and tumor necrosis factor-alpha (TNF-alpha) levels, resuscitation fluid requirements, and histologic mucosal injury were evaluated in all groups. RESULTS: The RAAA model resulted in increased lung and intestinal permeability to radiolabeled albumin and lung MPO activity (P <.01), with increases in intestinal TNF-alpha (P <.001) and CINC (P <.01) levels, when compared with sham-operated animals. Treatment with anti-rat CINC monoclonal antibody attenuated the increases in intestinal permeability and histologic mucosal injury (P <.01), gut TNF-alpha level (P <.001), and resuscitation fluid volume required (P <.05), without significantly affecting lung and intestinal MPO activity, lung permeability, and intestinal CINC level (P = NS), compared with animals given saline solution. CONCLUSION: Neutralization of CINC by the anti-rat CINC monoclonal antibody attenuated intestinal injury and induction of intestinal TNF-alpha, but failed to significantly attenuate remote pulmonary injury in this model of RAAA. PMID- 15111869 TI - Aortic endograft thrombosis after colorectal surgery in lithotomy position. AB - Aortic endograft limb occlusion is a serious complication after endovascular abdominal aortic aneurysm repair. We describe a yet unreported cause of endograft limb occlusion, the lithotomy position. Two patients with abdominal aortic aneurysm and colorectal cancer underwent an initial endovascular repair followed by cancer resection in the lithotomy position. Aortic endograft limb occlusion occurred in both patients immediately after the cancer operation. Percutaneous rheolytic thrombectomy was performed successfully in both patients. Pelvic surgery requiring the lithotomy position should be performed with caution in patients with aortic endografts, because it can result in endograft occlusion. PMID- 15111870 TI - Total laparoscopic abdominal aortic aneurysm repair with reimplantation of the inferior mesenteric artery. AB - We performed a total laparoscopic reimplantation of the inferior mesenteric artery (IMA) during laparoscopic infrarenal aortic aneurysm repair. The postoperative course was uneventful, and angiograms showed a patent IMA after reimplantation. To our knowledge, total laparoscopic reimplantation of the IMA in human beings has not previously been described. PMID- 15111871 TI - Transluminal thrombin injection and exclusion of a paramesenteric abdominal aortic aneurysm. AB - Surgical repair of aortic aneurysms involving the visceral arteries carries high morbidity and mortality in poor surgical candidates. With current technology, visceral artery involvement generally precludes endovascular repair of aortic aneurysms. We report on a patient with a large abdominal aortic pseudoaneurysm involving the origin of the superior mesenteric artery. This aneurysm was successfully repaired by transluminal thrombin injection of the sac and exclusion with balloon expandable covered stents placed in the aorta. PMID- 15111872 TI - Coil embolization of multiple hepatic artery aneurysms in a patient with undiagnosed polyarteritis nodosa. AB - Hepatic aneurysms are a rare sequela of vascular abnormalities in the liver, including trauma, infection, necrotizing vasculitis such as polyarteritis nodosa (PAN), and iatrogenic and arterial mediolysis. Presentation with intra-abdominal hemorrhage is associated with a high mortality rate. We describe life-saving transcatheter coil embolization of multiple isolated ruptured hepatic pseudoaneurysms in a patient with no history or clinical findings of PAN. We present angiographic findings and intra-arterial transcatheter embolization techniques in the treatment of ruptured large hepatic artery aneurysms. Endovascular specialists should recognize that PAN could present with classic angiographic findings and, in some cases, as life-threatening ruptured isolated hepatic artery aneurysms as its first presentation. PMID- 15111873 TI - Aneurysm of the femoral artery occupationally exposed to a vibratory tool for more than 10 years. AB - The first case of an aneurysm of the femoral artery occupationally exposed to a vibratory tool is described. A 72-year-old man with a right common femoral artery aneurysm had had an occupational history of putting a "breaker," which breaks concrete into pieces by means of powerful vibration, on the right groin for more than 10 years. The patient underwent aneurysmectomy and graft replacement, with reconstruction of the deep femoral artery. Pathologic examination of the resected aneurysm revealed fibrosis and lipid deposition in the intima, well-maintained elastic fibers without disruption in the media, and thickened adventitia, in place of the typical findings of atherosclerosis. PMID- 15111874 TI - Endovascular repair of a ruptured thoracic aortic aneurysm with the use of aortic extension cuffs. PMID- 15111875 TI - Current status of open versus endovascular stent-graft repair of abdominal aortic aneurysm. PMID- 15111876 TI - Ethics of professional courtesy. PMID- 15111893 TI - Entrapment of the long head of the biceps tendon: the hourglass biceps--a cause of pain and locking of the shoulder. AB - We describe an unrecognized mechanical condition affecting the long head of the biceps (LHB) tendon with entrapment of the tendon within the joint and subsequent pain and locking of the shoulder on elevation of the arm. We identified 21 patients with a hypertrophic intraarticular portion of the LHB tendon during open surgery (14 patients) or arthroscopic surgery (7 patients). All cases but one were associated with a rotator cuff rupture. Patients were treated by biceps tenotomy (2 patients) or tenodesis (19 patients) after removal of the hypertrophic intraarticular portion of the tendon and appropriate treatment of concomitant lesions. Minimum follow-up was 1 year. All patients presented with anterior shoulder pain and loss of active and passive elevation averaging 10 degrees to 20 degrees. A dynamic intraoperative test, involving forward elevation with the elbow extended, demonstrated entrapment of the tendon within the joint in each case. This test creates a characteristic buckling of the tendon and squeezing of it between the humeral head and the glenoid (hourglass test). The mean Constant score improved from 38 to 76 points at the final follow-up (P <.05). Complete and symmetric elevation was restored in all cases after resection of the intraarticular portion of the LHB tendon. The hourglass biceps is caused by a hypertrophic intraarticular portion of the tendon that is unable to slide into the bicipital groove during elevation of the arm; it can be compared with the condition of trigger finger in the hand. A loss of 10 degrees to 20 degrees of passive elevation, bicipital groove tenderness, and radiographic findings of a hypertrophied tendon can aid in the diagnosis. A definitive diagnosis is made at surgery with the hourglass test: incarceration and squeezing of the tendon within the joint during forward elevation of the arm with the elbow extended. The hourglass biceps is responsible for a mechanical block, which is similar to a locked knee with a bucket-handle meniscal tear. Simple tenotomy cannot resolve this mechanical block. Excision of the intraarticular portion of the LHB tendon, during bipolar biceps tenotomy or tenodesis, must be performed. The hourglass biceps is an addition to the familiar pathologies of the LHB (tenosynovitis, prerupture, rupture, and instability) and should be considered in cases of shoulder pain associated with a loss of elevation. PMID- 15111894 TI - A prospective, double-blind comparison of magnetic resonance imaging and arthroscopy in the evaluation of patients presenting with shoulder pain. AB - The purpose of this study was to determine the diagnostic ability of magnetic resonance imaging (MRI) compared with a reference standard, arthroscopy, in patients presenting with shoulder pain consistent with the signs and symptoms of shoulder impingement. Fifty-eight patients in whom conservative management failed underwent standardized MRI with intravenous gadolinium enhancement and arthroscopic evaluation. The prevalence of supraspinatus tendon, long head of the biceps tendon, and acromioclavicular joint pathology in this population was high: 79.3%, 66%, and 56%, respectively. MRI was highly accurate in detecting full thickness supraspinatus tears and acromioclavicular joint pathology. However, it had poor concordance with arthroscopy in diagnosing pathology in the biceps tendon and in classifying the curvature of the acromion. A smaller number of other abnormal structures were identified in this population of patients, including labral abnormalities (superior labral anterior-posterior and Bankart tears), Hill-Sachs lesions, and articular surface damage. Concordance between MRI and arthroscopy would increase with improved radiologist-surgeon communication, a more reliable classification of pathology, and possibly the use of intraarticular contrast. This study demonstrates that patients presenting with the clinical findings of shoulder impingement syndrome represent a heterogeneous population. Overall, MRI is a useful tool in the identification of shoulder pathology; however, the clinical correlation of this information and the assessment of outcomes remains unknown. PMID- 15111895 TI - Cementless surface replacement arthroplasty (Copeland CSRA) for osteoarthritis of the shoulder. AB - Clinical experience with the Copeland cementless surface replacement arthroplasty (CSRA) of the shoulder now spans 17 years. Between 1986 and 1997, 79 CSRAs (42 total shoulder replacements and 37 hemiarthroplasties) were performed for primary osteoarthritis of the shoulder. Total shoulder replacement was done in 12 men and 30 women with a mean age of 71.5 years (range, 50-87 years). Hemiarthroplasty was used in 9 men and 28 women; 5 patients had bilateral hemiarthroplasty. The mean age was 73.4 years (range, 53-88 years). Thirty-nine total shoulder arthroplasties and thirty hemiarthroplasties with a follow-up of more than 2 years were available for review. The mean follow-up was 7.6 years (range, 48 months to 13 years) for total shoulder replacement and 4.4 years (range, 24 months to 6.5 years) for hemiarthroplasty. The Constant scores improved from an age-adjusted Constant score of 33.8% (20.0 points) to 94% (61.9 points) for total shoulder replacement and from an age-adjusted Constant score 40.0% (25.3 points) to 91% (58.1 points) for hemiarthroplasty. Active elevation improved by a mean of 59.9 degrees to a mean of 128 degrees for total shoulder replacement and to a mean of 124 degrees for hemiarthroplasty. Of the patients, 89.9% considered the shoulder to be much better or better as a result of the operation. Radiographically, one humeral implant and three glenoid implants had evidence of loosening. Four revisions were performed in the total shoulder replacement group. No revision surgery was needed in the hemiarthroplasty group. The results of this series are at least comparable to those reported for stemmed prostheses with a comparable length of follow-up. The results of total shoulder replacement and hemiarthroplasty in osteoarthritis of the shoulder seem to be comparable. With use of the CSRA prosthesis, several severe complications mainly concerning the humeral shaft and periprosthetic fractures can be avoided. Should the need for revision surgery or arthrodesis arise, these procedures are easily performed, as bone stock has been maintained and no loss of length has been encountered. It does seem that the humeral component does not need a stem or cement for fixation. PMID- 15111896 TI - Reversed arthroscopic subacromial decompression for massive rotator cuff tears. AB - This prospective study evaluates the results of a procedure for massive rotator cuff tears that we term reversed arthroscopic subacromial decompression (ASD). The procedure includes an arthroscopic debridement of the subacromial space and glenohumeral joint, an arthroscopic tuberoplasty, and depending on the pathologic condition of the long head of the biceps, a biceps tendon tenotomy. Reversed ASD avoids a classic acromioplasty in order to preserve the integrity of the coracoacromial arch. Twenty-three patients with a mean age of 69 years underwent this procedure. After a mean follow-up of 40 months, the age-adjusted Constant score increased significantly, from 65.9% to 90.6% (P <.001), with 14 excellent, 5 good, 2 satisfactory, and 1 poor result. Preexisting osteoarthritic changes increased significantly but had no impact on the final clinical results. The acromiohumeral distance decreased from 5.1 to 4.5 mm (P =.004). There were no complications directly related to the surgical procedure. When compared with classic ASD studies for massive rotator cuff tears, we obtained similar midterm results with regard to pain relief, functional recovery, and patient satisfaction. We, therefore, conclude that reversed ASD with tenotomy of the long head of the biceps tendon offers a less invasive treatment strategy for massive rotator cuff tears while preserving the integrity of the coracoacromial arch. PMID- 15111897 TI - Reasons for failure after surgical repair of anterior shoulder instability. AB - A total of 41 patients presenting with recurrent anterior instability of the shoulder after surgical repair were followed up after a mean period of 49 months (range, 24-81 months). The failed procedures were arthroscopic Bankart repair in 25 cases, open Bankart repair in 6 cases, Eden-Hybinette procedure in 4 cases, rotational osteotomy in 2 cases, capsular T- shift operation in 1 case, Bristow Latarjet in one case, and a J-bone graft procedure in one case. In one case the index procedure was unknown. At revision surgery, the findings were a defect of the anterior bony glenoid rim in 23 patients (56%), a large capsule in 9 (22%), and a laterally torn capsule in 2 (5%). In 7 patients (17%) a typical Bankart lesion with good capsule quality was found. At revision surgery, these lesions were addressed by a bone graft procedure in 21 cases and fixation of the rim fragment with screws in 2 cases. In the 9 patients with a large capsule, a T shift operation was performed in 6 and a Bankart repair with capsulorrhaphy was performed in the remaining 3. In the 7 patients with a typical Bankart lesion, a Bankart repair was performed, and in the 2 patients with a laterally torn capsule, an open suturing technique was used. At follow-up, none of the patients had had further redislocation or subluxation. The Rowe score was excellent in 81% of the cases and good in 19%. In 19 patients (46%) no increase in arthritic change was detected on radiographic assessment at follow-up, whereas 13 (32%) showed an increase of 1 degree and 4 (10%) showed an increase of 2 degrees. The results show that good and very good outcomes can be achieved with surgical repair provided that the basic pathology of the unstable shoulder is taken into account. PMID- 15111898 TI - Ultrasonographic evaluation of the Bankart lesion. AB - The Bankart lesion is an essential finding of traumatic anterior shoulder instability. The purpose of this study was to clarify the reliability of diagnosis by ultrasonography (US) of the Bankart lesion when using an axillary approach. Six cadaveric shoulders were examined by US from the axilla. The shoulders were then dissected along the plane of the ultrasonic beam. The labrum at the 4-o'clock position of all 6 cadavers was present along the ultrasonic plane; there were no obstacles from the skin to the glenoid rim, and all cases had a normal labrum at the 4-o'clock position. US images of the normal labrum showed that the labrum was continuously connected to the glenoid rim with no borderline (low-echo line) between them. Preoperative US from the axilla was performed in 75 patients who underwent arthroscopic surgery. Anteroinferior labra were classified preoperatively from the US images as normal, detached (low-echo line between glenoid rim and labrum), or displaced (labrum displaced from glenoid rim), with a diagnosis of a Bankart lesion being made for the latter two types. US findings were correlated with arthroscopic observations. This revealed that for the detection of Bankart lesions, US had a sensitivity of 88.6%, a specificity of 77.4%, an accuracy of 84.0%, a positive predictive value of 84.8%, and a negative predictive value of 82.8%. These results demonstrate that US with an axillary approach is useful in the diagnosis of Bankart lesions of the shoulder. PMID- 15111899 TI - The use of in-office, orthopaedist-performed ultrasound of the shoulder to evaluate and manage rotator cuff disorders. AB - This study presents the use of in-office ultrasound, performed by an attending orthopaedic surgeon, as a means of evaluating the integrity of the rotator cuff. The results of 282 shoulder sonograms in patients ultimately treated surgically were included. Findings at surgery were recorded and compared with those documented during the ultrasound examination. Ultrasound findings included 118 full-thickness and 143 partial-thickness rotator cuff tears and 6 intact cuffs confirmed at surgery. One patient with a partial supraspinatus tear on ultrasound was intact at surgery, nine with complete supraspinatus tears had partial thickness tears at surgery, one with an intact supraspinatus had a full-thickness tear at surgery, and four with partial-thickness supraspinatus tears had full thickness tears at surgery. The sensitivity, specificity, positive predictive value, and negative predictive value were 94.1%, 96.1%, 96.6%, and 93.2%, respectively, for partial-thickness tears; 95.9%, 94.3%, 92.9%, and 96.8%, respectively, for full-thickness tears; and 99.6%, 85.7%, 99.6%, and 85.7%, respectively, when the rotator cuff was evaluated for damage (either partial- or full-thickness tears). This series documents the ability of an orthopaedic surgeon to image the rotator cuff effectively using portable ultrasound in the clinic setting, allowing for a more efficient implementation of the management plan. PMID- 15111900 TI - Evaluation of the lag sign tests for external rotator function of the shoulder. AB - Rotator cuff lesions pose a serious clinical challenge. The objective of this study was to verify the biomechanical basis for the lag sign clinical tests for rotator cuff dysfunction. The lag sign tests were simulated in vitro by a sensor guided robotic simulator configured to reproduce in vivo testing conditions. The ability of the test to isolate supraspinatus and/or infraspinatus dysfunction was investigated from 20 degrees to 90 degrees of scapular plane arm elevation. The test was 100% sensitive for lack of all infraspinatus-teres minor force at all elevations tested (6/6 specimens at 20 degrees, 30 degrees, and 60 degrees elevation; 5/5 at 90 degrees elevation). The test was less sensitive to complete loss of supraspinatus force (1/6 specimens testing positive at 20 degrees, 0/6 at 30 degrees, 3/6 at 60 degrees, and 3/5 at 90 degrees elevation). The results of this biomechanical study suggest the lag sign tests to be highly sensitive only for infraspinatus-teres minor muscle dysfunction. PMID- 15111901 TI - Shoulder arthroplasty for late sequelae of proximal humeral fractures. AB - Twenty-eight patients with sequelae of proximal humeral fractures were treated with shoulder arthroplasty and were reviewed with a mean follow-up of 47 months. There were 8 malunions, 7 osteonecrosis, and 2 nonunions of the proximal humerus. Degenerative joint disease without any distortion of the tuberosities had developed in 11. We performed 8 total shoulder arthroplasties and 20 hemiarthroplasties. On the basis of the Neer criteria, the results were satisfactory in only 64%. Fifteen patients had superior migration of the implant. One patient had to be reoperated on because of deep infection. The prognosis was influenced positively by the integrity of the rotator cuff at surgery, whereas the need for greater tuberosity osteotomy worsened the final result. The data suggest that malunion of the greater tuberosity can be tolerated if it does not compromise acceptable positioning of the humeral component. However, if there is a malunion of the greater tuberosity with major displacement, an osteotomy must be performed, with unpredictable results. PMID- 15111902 TI - Total elbow arthroplasty with massive composite allografts. AB - Total elbow arthroplasty is challenging when there is extensive bone loss. We studied retrospectively the long-term (mean, 6.5 years) clinical and radiographic results of 10 patients in whom 14 massive allograft-prosthetic composites were used in such clinical situations. The mean arc of active flexion/extension was 92 degrees. The Bryan-Morrey and Hospital for Special Surgery elbow scoring systems were used and revealed a modest improvement in pain and stability but a disappointing return of function, with independent self-care possible in only 3 patients, household and employment activities in 2, and recreational pursuits in 2. Three patients claimed that they were unable to use the elbow. Multiple procedures (mean, 2.2) were sometimes required to optimize the result. The 79% rate of allograft-host union is similar to that of hip and knee arthroplasty. The use of massive allografts is a reasonable alternative in salvage situations involving total elbow arthroplasty with massive bone loss. PMID- 15111903 TI - A cadaveric study on the anatomy of the deltoid insertion and its relationship to the deltopectoral approach to the proximal humerus. AB - Elevation of the deltoid insertion (DI) has been recommended, but little is known about its anatomy or importance for deltoid function. The purpose of this study is to determine the dimensions of the DI with specific reference to the deltopectoral approach. The deltoid was exposed and detached at its origin in 36 cadaveric shoulders. The morphology of the DI was documented, and its relationship with the pectoralis major insertion and the axillary and radial nerves was recorded. The anterior, middle, and posterior deltoid muscle fibers entered into the DI in a V-shaped tendinous confluence with a broad posterior band and a narrow separate anterior band, which accounted for the anterior one fifth of the DI (0.44 cm). The deltoid insertion was separated from the pectoralis major insertion by as little as 2 mm in 31 of 36 specimens. The distance between the axillary nerve and the DI averaged 5.6 cm anteriorly and 4.5 cm posteriorly. The distance between the radial nerve and posterior deltoid insertion averaged 2.4 cm proximally and 1.6 cm distally. Exposure during the deltopectoral approach is most limited by the close proximity of the deltoid and pectoralis major insertions. Our study would suggest that partial anterior DI release (greater than one fifth) could compromise the anterior deltoid. The axillary and radial nerves are not at significant risk when operating in the region of the anterior DI. PMID- 15111904 TI - Fibrous cartilage in the rotator cuff: A pathogenetic mechanism of tendon tear? AB - There is no consensus on the pathogenesis of rotator cuff tears. Fibrous cartilage has been hypothesized to develop in some tendons as a result of shear or compressive forces, resulting in a tissue less capable of resisting normal tensile load and more prone to tearing. To test the hypothesis that metaplastic fibrocartilage in the rotator cuff could be involved in the pathogenic mechanism of its tear, samples from 34 acute and chronic torn rotator cuffs were subjected to histologic and immunohistologic study for the presence and type of cartilage (hyaline, fibrous, or elastic) in the area of the lesion and surrounding tissues. Detection of type I and II collagen, S-100 protein, and chondroitin sulfate allowed areas of fibrous cartilage to be seen in all samples, suggesting that the characteristic of rotator cuff tendons to work both in tension and in compression may stimulate fibrocartilaginous metaplasia and lead to a complete tear. PMID- 15111905 TI - Association of osteopenia of the humeral head with full-thickness rotator cuff tears. AB - Rotator cuff tendon repair may fail for various reasons. Although the role of repair techniques and of the musculotendinous unit has been studied, there is little information on the quality of the bone to which the tendon is to be repaired. Therefore, 14 cadaveric humeral heads, 7 specimens without and 7 with a full-thickness rotator cuff tendon tear, were quantitatively assessed by use of high-resolution micro-computed tomography. Bone density is higher below the articular surface than in the greater tuberosity (40% vs 10%-20%), and tendon tears are associated with a reduction in cancellous bone density of greater than 50%, leading to a virtually hollow greater tuberosity, with intact cortical bone. The results found suggest that in long-standing rotator cuff tears, creating a deep trough should be avoided to achieve reliable tendon-to-bone contact. For optimal suture fixation to bone, sutures or anchors should be positioned subcortically or medially under the articular surface. PMID- 15111906 TI - Effect of subacromial decompression on laxity of the acromioclavicular joint: biomechanical testing in a cadaveric model. AB - Subacromial decompression is a well-accepted treatment for impingement syndrome when nonoperative therapies have failed. However, recent clinical data have raised concern that arthroscopic subacromial decompression may lead to laxity of the acromioclavicular joint and, potentially, predispose patients to late postoperative acromioclavicular joint pain. Our goal was to determine whether subacromial decompression with co-planing of the distal clavicle alters the laxity, or compliance, of the acromioclavicular joint in a cadaveric model. Eighteen cadaveric shoulders were dissected and tested in a specially designed rig, driven by a hydraulic materials testing machine. One hundred-Newton loads were applied to the distal clavicle in the superior, posterior, and anterior directions, while acromioclavicular joint motion was recorded with a 3 dimensional infrared optical measurement system. Acromioplasty was performed with a posterior-referenced cutting block technique and included co-planing of the distal clavicle in all specimens. Joint compliance before and after subacromial decompression was compared with the paired t test. Subacromial decompression increased anteroposterior compliance by 13%, from 8.8 +/- 2.9 mm (mean +/- SD) in the intact joint to 9.9 +/- 3.1 mm (P =.001). Subacromial decompression increased superior compliance by 32%, from 3.1 +/- 1.5 mm in the native specimen to 4.1 +/- 1.8 mm (P =.03). These observations may have implications for the technique of acromioplasty. Although the immediate result of acromioplasty with co-planing appears to be an increase in the compliance of the acromioclavicular joint, the clinical significance of these findings has yet to be determined. PMID- 15111907 TI - Total, trabecular, and cortical bone mineral density in different regions of the glenoid. AB - The purpose of this study was to analyze total, trabecular, and cortical bone mineral density (BMD) of different regions of the glenoid. Soft tissue was dissected from 20 cadaveric shoulders, and the glenoids were isolated. Axial peripheral quantitative computed tomography (pQCT) scans perpendicular to the articular surface were performed at the middle level of each (superior, middle, and inferior) horizontal third of the glenoid, and the total, trabecular, and cortical BMDs of the anterior and posterior half of each region were determined by use of pQCT software. The total BMDs of the posterior and superior glenoid were significantly higher than those of the anterior and inferior glenoid (P <.01). Trabecular BMD of the posterior glenoid was significantly higher than that of the anterior glenoid, and cortical BMD of the superior glenoid was significantly higher than that of the inferior glenoid (P <.01). These significant differences in distribution of the BMD explain previous findings of the mechanical properties of the glenoid. Future glenoid component design should provide maximal surface area for posterior and superior bone ingrowth to the component. Posteriorly and superiorly, the glenoid bone stock also provides stronger support for any kind of fixation on the bony surface. PMID- 15111908 TI - Valgus torque in youth baseball pitchers: A biomechanical study. AB - The purpose of this study was to determine the biomechanical and anthropometric factors contributing to elbow valgus torque during pitching. Video data of 14 youth pitchers throwing fastballs were used to calculate shoulder and elbow kinematics and kinetics. Peak elbow valgus torque averaged 18 Nm and occurred just before maximal shoulder external rotation. The magnitude of valgus torque was most closely correlated with the thrower's weight. When subject weight and height were controlled for, maximum shoulder abduction torque and maximum shoulder internal rotation torque were most strongly associated with elbow valgus torque, accounting for 85% of its variance (P <.001). When only kinematic variables were considered, maximum shoulder external rotation accounted for 33% of the variance in valgus torque. Given that the biomechanical variables correlated with peak valgus torque are not easily modifiable, limiting the number of innings pitched is likely the best way to reduce elbow injury in youth pitchers. PMID- 15111909 TI - Unrecognized fracture of the medial epicondylar apophysis of the humerus. PMID- 15111910 TI - Anterior dislocation of the shoulder with rotator cuff injury and brachial plexus palsy: a case report. PMID- 15111911 TI - Case report: Management of occult tuberculosis infection by 2-stage arthroplasty of the elbow. PMID- 15111912 TI - Locked intramedullary nailing in the treatment of olecranon nonunion: a new method of treatment. PMID- 15111913 TI - Health-related quality of life predicts emergency department utilization for patients with asthma. AB - STUDY OBJECTIVE: The objective of this study is to evaluate health-related quality of life as a risk factor for subsequent emergency department (ED) utilization in a cohort of patients with asthma. METHODS: We conducted a 1-year prospective cohort study of 1,406 adult asthma patients. Baseline physical and mental health status were measured using the Physical Component Summary and Mental Component Summary scores from the Short Form-12 health status survey. Asthma-specific health-related quality of life was measured with the mini-Asthma Quality of Life Questionnaire. Multivariable regression was used to assess the independent association between baseline Physical Component Summary, Mental Component Summary, and Asthma Quality of Life Questionnaire scores and asthma related ED visits during the subsequent year. RESULTS: During the 1-year follow up period, 116 patients made at least 1 asthma-related ED visit. After adjustment for multiple sociodemographic and clinical factors, both the Physical Component Summary score (odds ratio [OR] 1.72; 95% confidence interval [CI] 1.46 to 2.02) and the Asthma Quality of Life Questionnaire score (OR 1.34; 95% CI 1.18 to 1.52) were associated with subsequent asthma-related ED utilization. In contrast, overall mental health status was not associated with subsequent asthma-related ED utilization (OR 1.17; 95% CI 0.96 to 1.44). CONCLUSION: Overall physical health status and asthma-specific quality of life predict subsequent ED utilization. Health-related quality of life may be useful in identifying patients at increased risk for asthma exacerbation requiring emergency care. PMID- 15111914 TI - The impact and enforcement of prudent layperson laws. AB - STUDY OBJECTIVE: Almost every state has enacted a "prudent layperson" standard for determining insurance coverage for emergency department (ED) services. This study evaluates whether these laws are achieving their goals or causing unintended side effects. METHODS: Six states were selected for in-depth case studies to represent a range of market, demographic, and legal conditions. In each state, 11 to 15 interviews were conducted with insurers, regulators, providers, employers, patient advocates, and industry observers, for a total of 87 interviews. In addition, regulators in all 50 states completed a written survey about likely enforcement responses for hypothetical violations of these laws. RESULTS: Basic compliance with prudent layperson laws appears to be widespread. Regulators actively enforce these laws, and most subjects reported no systematic violations. Insurers explained that it is difficult to operationalize a coverage standard that relies on patients' experience of symptoms rather than on providers' assignment of diagnostic and procedure codes. No strong evidence was found that these laws have significantly increased insurance costs, which is due in part to various strategies insurers have adopted to reduce payments to providers for ED services and to greatly increase patients' copayments. Accordingly, few subjects believe these laws have increased inappropriate ED use. CONCLUSION: Prudent layperson laws have helped to catalyze industry-wide changes in how health insurers review ED claims and how they manage ED costs. Whether these changes, on balance, are beneficial to patients and to society requires further study focused on outcomes and system-wide costs. PMID- 15111915 TI - Access to emergency care: restricted by long waiting times and cost and coverage concerns. AB - STUDY OBJECTIVE: We monitor progress toward Healthy People 2010 objectives of reducing health disparities and decreasing delay and difficulty in access to emergency care. METHODS: This was a secondary analysis of 2001 National Health Interview Survey interviews of 33,326 adults to provide population-based estimates of self-reported delay, difficulty, or inability to get care from a hospital emergency department (ED) in the preceding 12 months. RESULTS: About 7.7% of the estimated 36.6 million adults who sought care in a hospital ED in the preceding 12 months reported a delay in receiving care, having difficulty receiving care, or being unable to receive care. Waiting times were the most frequently noted cause of problems. Concerns about service costs and insurance coverage were also commonly cited access barriers. Access problems were more likely to be reported by adults without health insurance, younger adults, adults in fair or poor health, and adults with annual incomes of less than 20,000 dollars. CONCLUSION: Self-reported access to ED care is impeded by prolonged waiting times and by cost and insurance coverage concerns. These access problems are occurring more frequently among groups that face multiple social and economic disadvantages. Hospital operational changes to reduce ED treatment delays and health care financing policies that reduce insurance coverage inequities may both be needed to meet these Healthy People 2010 objectives. PMID- 15111916 TI - Clinical features and management of herb-induced aconitine poisoning. AB - STUDY OBJECTIVE: We define the potential sources, clinical manifestations, and treatment of aconitine poisoning. METHODS: The database of the National Poison Center in Taiwan was retrospectively searched for the diagnosis of aconitine poisoning for 1990 to 1999. The reasons for taking the aconite roots, the clinical features, management, and possible predisposing factors were noted. RESULTS: A total of 17 cases occurred and consisted of 9 men and 8 women aged 30 to 70 years. Thirteen patients ingested aconite roots as treatment for rheumatism and wounds. Two patients volunteered to test the effects of aconite roots in a drug study. Two patients accidentally ingested the aconite roots. After a latent period of 10 to 90 minutes, patients developed a combination of neurologic (n=17), cardiovascular (n=14), gastrointestinal (n=9), and other (n=5) features typical of aconitine poisoning. Four patients developed ventricular tachycardia. All patients received supportive treatment. Patients with ventricular tachycardia were also treated with charcoal hemoperfusion. All patients made a complete recovery. CONCLUSION: Life-threatening ventricular tachycardia can occur after the consumption of aconite roots. The risk is higher with inadequately processed aconite roots, large doses, or tincture preparations. With increasing popularity of herbal medicines, herb-induced aconitine poisoning may also be seen in Western countries. PMID- 15111917 TI - Buprenorphine: a primer for emergency physicians. AB - The recent approval of office-based treatment for opioid addiction and US Food and Drug Administration approval of buprenorphine will expand treatment options for opioid addiction. Buprenorphine is classified as a partial micro opioid agonist and a weak kappa antagonist. It has a high affinity for the micro receptor, with slow dissociation resulting in a long duration of action and an analgesic potency 25 to 40 times more potent than morphine. At higher doses, its agonist effects plateau and it begins to behave more like an antagonist, limiting the maximal analgesic effect and respiratory depression. This "ceiling effect" confers a high safety profile clinically, a low level of physical dependence, and only mild withdrawal symptoms on cessation after prolonged administration. Suboxone contains a mixture of buprenorphine and naloxone. The naloxone is poorly absorbed sublingually and is designed to discourage intravenous use. Subutex, buprenorphine only, will also be available primarily as an initial test dose. Clinicians will be using this drug for detoxification or for maintenance of opioid addiction. Patients with recent illicit opioid use may develop a mild precipitated withdrawal syndrome with the induction of buprenorphine. Acute buprenorphine intoxication may present with some diffuse mild mental status changes, mild to minimal respiratory depression, small but not pinpoint pupils, and relatively normal vital signs. Naloxone may improve respiratory depression but will have limited effect on other symptoms. Patients with significant symptoms related to buprenorphine should be admitted to the hospital for observation because symptoms will persist for 12 to 24 hours. PMID- 15111918 TI - The relationship of intraocular pressure to intracranial pressure. AB - STUDY OBJECTIVE: The early detection of intracranial hypertension can lead to timely medical and neurosurgical intervention, preventing brain herniation and death. In this investigation, we hypothesize that an increase in intracranial pressure can be detected by an increase in intraocular pressure using noninvasive existing technology, the handheld tonometer. METHODS: This was a prospective observational pilot study conducted at a community hospital. Admitted patients with an invasive intracranial pressure monitor were solicited for participation. Patients were excluded if they had known glaucoma or had sufficient ocular or facial trauma that precluded intraocular pressure determination. Simultaneous measurements of intracranial and intraocular pressure were recorded. RESULTS: Twenty-seven patients were enrolled, and 76 individual measurements were performed. All patients with an abnormal intracranial pressure had an abnormal intraocular pressure; similarly, all patients with a normal intracranial pressure had a normal intraocular pressure (sensitivity 1.00, 95% confidence interval 0.86 to 1.0; specificity 1.0, 95% confidence interval 0.93 to 1.0) CONCLUSION: Abnormal intraocular pressure as measured with the handheld tonometer is an excellent indicator of abnormal intracranial pressure in patients with known intracranial pathology. PMID- 15111919 TI - Transient ischemic attack: review for the emergency physician. AB - In the past few years, the approach to patients with transient ischemic attacks has undergone a transformation. To care for these patients, emergency physicians must understand these changes. They must be comfortable with the diagnosis and treatment of transient ischemic attacks in their emergency department. To this end, we ask and answer the following 6 important questions in this up-to-date review of transient ischemic attacks: (1) How is a transient ischemic attack defined? (2) Does this patient have a transient ischemic attack? (3) Once diagnosed, what diagnostic evaluation should be done (and when)? (4) What treatment should be instituted (and when)? (5) What is the correct disposition? and (6) What are the current medical guidelines? PMID- 15111920 TI - Clinical policy: Critical issues in the evaluation and management of adult patients presenting to the emergency department with seizures. AB - This clinical policy focuses on critical issues in the evaluation and management of adult patients with seizures. The medical literature was reviewed for articles that pertained to the critical questions posed. Subcommittee members and expert peer reviewers also supplied articles with direct bearing on this policy. This clinical policy focuses on 6 critical questions: What laboratory tests are indicated in the otherwise healthy adult patient with a new-onset seizure who has returned to a baseline normal neurologic status?Which new-onset seizure patients who have returned to a normal baseline require a head computed tomography (CT) scan in the emergency department (ED)?Which new-onset seizure patients who have returned to normal baseline need to be admitted to the hospital and/or started on an antiepileptic drug?What are effective phenytoin or fosphenytoin dosing strategies for preventing seizure recurrence in patients who present to the ED after having had a seizure with a subtherapeutic serum phenytoin level?What agent(s) should be administered to a patient in status epilepticus who continues to seize after having received benzodiazepine and phenytoin?When should electroencephalographic (EEG) testing be performed in the ED? Recommendations for patient management are provided for each 1 of these topics on the basis of strength of evidence (Level A, B, or C). Level A recommendations represent patient management principles that reflect a high degree of clinical certainty; Level B recommendations represent patient management principles that reflect moderate clinical certainty; and Level C recommendations represent other patient management strategies based on preliminary, inconclusive, or conflicting evidence, or based on consensus of the members of the Clinical Policies Committee. This clinical policy is intended for physicians working in hospital based EDs. PMID- 15111921 TI - Acute endotracheal tube occlusion caused by use of an esophageal detector device: report of a case and a discussion of its utility. AB - An experienced intensivist performed an emergency endotracheal intubation on a 72 year-old woman without difficulty. The vocal cords were visualized during the intubation. An esophageal detector device was used to confirm correct airway placement. After aspiration of the device, it was impossible to easily ventilate the patient. The endotracheal tube was removed and found to be occluded by a mucus plug. It is postulated that aspiration of the syringe on the esophageal detector device suctioned the mucus plug from the bronchial tree into the endotracheal tube and precluded ventilation of the airway. This complication has not been previously reported. The utility and possible hazards of the esophageal detector device are discussed. PMID- 15111922 TI - Delayed obstruction of endotracheal tubes by aspirated foreign bodies: report of two cases. AB - We present 2 cases of endotracheal tube obstruction as a result of previously aspirated foreign bodies that moved from the bronchial tree into the endotracheal tube. The signs of endotracheal tube obstruction were demonstrated in both cases: (1) activation of the high-pressure alarm; (2) difficulty ventilating by using a bag-valve-mask device; (3) rapid decrease of end-tidal CO2; (4) oxygen desaturation that may have lagged behind the decrease of end-tidal CO2; and (5) the inability to advance a catheter down the endotracheal tube. The differential diagnosis of endotracheal tube obstruction is discussed, as are possible interventions to remove the obstruction and secure an open airway. PMID- 15111923 TI - Report of the Task Force on Residency Training Information (2003-2004), American Board of Emergency Medicine. AB - The American Board of Emergency Medicine gathers extensive background information on emergency medicine residents and the programs in which they train. We present the seventh annual report on the status of US emergency medicine residency programs. PMID- 15111924 TI - Career options in aerospace and aviation medicine. PMID- 15111925 TI - HIPAA enhancements to improve emergency department security. AB - The Health Insurance Portability and Accountability Act (HIPAA) seems to be as useful as is capitalism to the medically uninsured (or perhaps, as necessary as another leukotriene inhibitor for asthma). Is the emergency medical community doing enough? Can we improve on HIPAA to increase privacy in the emergency department? HIPAA regulations are reviewed in all their wondrous complexity and simplified so that even your medical director can understand them. PMID- 15111926 TI - Update on emerging infections: News from the Centers for Disease Control and Prevention. Hepatitis a outbreak associated with green onions at a restaurant- Monaca, Pennsylvania, 2003. PMID- 15111928 TI - Cytomegalovirus prophylaxis and treatment after hematopoietic stem cell transplantation in Canada: a description of current practices and comparison with Centers for Disease Control/Infectious Diseases Society of America/American Society for Blood and Marrow Transplantation guideline recommendations. AB - Prevention and management of cytomegalovirus (CMV) disease after hematopoietic stem cell transplantation (HSCT) is critically important, but clinical practices have historically been heterogeneous. The Centers for Disease Control (CDC), as part of a larger clinical practice guideline initiative, has published evidence based recommendations, but their effect on clinical practice has not been assessed. A survey was sent to all Canadian HSCT program directors to describe current practices. Current practices were subsequently compared with CDC guideline recommendations and with a Canadian survey published before the CDC guidelines. When current practices did not conform to guideline recommendations, a literature review was performed to determine whether current practices were supported by evidence published after the CDC guidelines. The survey response rate was 100%. Variability in practices was observed in several aspects of patient care, including (1) indications for CMV-negative blood products, (2) surveillance tests used to detect CMV infection, (3) duration of surveillance testing, (4) duration of maintenance treatment after preemptive therapy was initiated, and (5) treatment of CMV disease. Overall adherence to guideline recommendations was good, especially when they were supported by high-quality data. However, deviations from guideline recommendations were observed: (1) most Canadian allogeneic programs used shorter courses of preemptive therapy; (2) prevention measures aimed at late CMV disease were not systematically applied at most allogeneic programs; and (3) most autologous HSCT programs did not administer preemptive therapy even in high-risk recipients. When deviations occurred, recent evidence supported current practices in some instances (shorter courses of preemptive therapy) but not in others (absence of strategies to prevent late CMV disease). Compared with practices in Canada before publication of the CDC guidelines, a higher proportion of programs used CDC-recommended surveillance tests and treatment for CMV infection. Current practices in Canada remain heterogeneous. Discrepancies between current practices and CDC guideline recommendations occurred in situations either in which practices had changed in response to recently published data or in which evidence supporting a recommendation was poor. These results suggest an urgent need for the development of well-designed clinical trials. Incorporation of recent data into updated guidelines may be appropriate. PMID- 15111929 TI - CD25 expression distinguishes functionally distinct alloreactive CD4 CD134 (OX40) T-cell subsets in acute graft-versus-host disease. AB - CD134 (OX40) is expressed on activated CD4(+) donor T cells in allogeneic stem cell transplant recipients with acute graft-versus-host disease. The data presented here reveal that differential expression of CD25 by CD4(+) CD134(+) T cells allows separation of these activated cells into 2 phenotypically and functionally distinct alloreactive T-cell subsets. These subsets exhibit distinct tissue associations, with CD4(+) CD134(+) CD25(-) T cells preferentially found in lymphoid tissues and CD4(+) CD134(+) CD25(+) T cells located in lymphoid tissues and inflamed extralymphoid tissues. The CD25(-) T-cell subset exhibited potent proliferative responses to both concanavalin A and allogeneic host leukocytes. By contrast, the CD25(+) T-cell subset proliferated minimally in response to either treatment and inhibited alloantigen-induced proliferation of the CD25(-) subset. Proliferative unresponsiveness associated with the CD25(+) T-cell subset did not extend to cytokine secretion. When stimulated with alloantigen, both CD4(+) CD134(+) T-cell subsets responded by secreting interferon-gamma and interleukin (IL)-10, and neither T-cell subset produced detectable levels of IL-2 or IL-4. Three-day treatment of the CD25(+) T-cell subset with IL-2 restored the proliferative responsiveness of these cells to host alloantigens, suggesting that the proliferative unresponsiveness associated with this T-cell subset reflected a requirement for IL-2. The preferential tissue associations and distinct functional properties associated with these separable alloreactive CD4(+) CD134(+) T-cell subsets suggest that they participate differentially in clinical graft-versus-host disease. PMID- 15111930 TI - Outcomes of adults with acute myelogenous leukemia in remission given 550 cGy of single-exposure total body irradiation, cyclophosphamide, and unrelated donor bone marrow transplants. AB - On the basis of observations from dog models and human studies, we hypothesized that a low-dose (550 cGy), single-exposure total body irradiation (TBI)-based regimen would result in improved survival when given to adult patients with acute myelogenous leukemia (AML) who were undergoing unrelated donor bone marrow transplantation in complete remission (CR). The regimen consisted of single exposure (550 cGy) of TBI given at a high dose rate (30 cGy/min) and cyclophosphamide. Graft-versus-host disease prophylaxis consisted of cyclosporine, methotrexate, and corticosteroids. Thirty-two consecutive adult patients (median age, 47 years) with AML in CR (15 in CR 1 and 17 in CR > or =2) were treated. Sixteen patients (50%) were alive and in remission at last follow up (median, 2.2 years; range, 0.6-4.0 years). Kaplan-Meier estimates of overall and leukemia-free survival at 3 years were 55% +/- 14% (mean +/- SE) and 57% +/- 14% in CR 1 patients and were both 39% +/- 12% in CR > or =2 patients. Transplant related mortality was 13% for patients in CR 1 and 41% for those in CR > or =2. Only 1 patient (3%) experienced fatal regimen-related organ toxicity, and only 1 had grade III or IV acute graft-versus-host disease. Graft failure was not observed. Relapse occurred in 22% of patients. This low-dose (550 cGy), single exposure TBI-based regimen resulted in good survival and a low risk of fatal regimen-related organ toxicity in adult patients with AML who underwent unrelated donor bone marrow transplantation in CR. PMID- 15111931 TI - Increasingly frequent diagnosis of acute gastrointestinal graft-versus-host disease after allogeneic hematopoietic cell transplantation. AB - The reported incidence of grades II to IV acute graft-versus-host disease (GVHD) after hematopoietic cell transplantation with HLA-identical sibling donors has increased considerably during the past 15 to 20 years at our center. The purpose of this study was to evaluate the potential reasons for this change. We reviewed organ stages and overall grades of GVHD for 2220 patients who received a first marrow or peripheral blood cell transplant from an HLA-identical sibling or an HLA-allele-matched unrelated donor with the use of a posttransplantation immunosuppressive regimen that included both methotrexate and cyclosporine between 1985 and 2001. The most striking change was an increased incidence of stage 1 gut involvement from 10% to 20% before 1992 to 50% to 60% since 1992, both with related and unrelated donors. This change increased the incidence of grade II GVHD with sibling donors, such that the overall incidence of grade II to IV GVHD is now 60% to 70%. Among patients with chronic myeloid leukemia in chronic phase, the increasingly frequent diagnosis of acute GVHD since 1992 has not been associated with decreased survival. A high diagnostic sensitivity and increased awareness that gut GVHD can occur without skin involvement account for the increased incidence of acute GVHD at our center. PMID- 15111932 TI - Sirolimus and tacrolimus without methotrexate as graft-versus-host disease prophylaxis after matched related donor peripheral blood stem cell transplantation. AB - Methotrexate in combination with a calcineurin inhibitor is a standard graft versus-host disease (GVHD) prophylactic regimen in allogeneic stem cell transplantation. However, methotrexate is associated with delayed engraftment, mucositis, idiopathic pneumonia syndrome, and other transplant-related complications. Sirolimus, a novel immunosuppressant without methotrexate's toxicities, has been used successfully in solid organ transplantation. We hypothesized that replacing methotrexate with sirolimus would preserve effective prophylaxis of GVHD while minimizing transplant-related toxicity after allogeneic peripheral blood stem cell transplantation. We enrolled 30 patients in a phase II study to test the efficacy of tacrolimus in combination with sirolimus in lieu of methotrexate in preventing GVHD after allogeneic peripheral blood stem cell transplantation from HLA-matched related donors. Grade II GVHD occurred in 3 patients (10%), and no patient developed grade III or IV GVHD. Neutrophil and platelet engraftment were prompt, occurring on days 14 and 13, respectively. All patients survived to hospital discharge (median, 18 days), and peritransplantation toxicity was mild. Four patients developed thrombotic microangiopathy, and 3 patients developed hepatic veno-occlusive disease. Chronic GVHD occurred in 11 patients. Relapse-free and overall survival at 100 days were 93% and 97%, respectively, and were 71% and 67% at 1 year. Causes of death included relapse (n = 6), veno-occlusive disease (n = 1), and late pulmonary toxicity (n = 1). Sirolimus in combination with tacrolimus is a promising alternative to methotrexate-based regimens for GVHD prophylaxis after matched related donor peripheral blood stem cell transplantation. Mucositis was modest, engraftment was prompt, and transplant-related toxicity was modest. Methotrexate free, sirolimus-based GVHD prophylactic regimens should be tested in randomized trials against the current standard of care. PMID- 15111933 TI - Chimerism studies in HLA-identical nonmyeloablative hematopoietic stem cell transplantation point to the donor CD8(+) T-cell count on day + 14 as a predictor of acute graft-versus-host disease. AB - Chimerism analysis of hematopoietic cells has emerged as an essential tool in nonmyeloablative hematopoietic stem cell transplantation. We have investigated the development of donor chimerism in granulocytes and CD4(+) and CD8(+) T cells in blood and bone marrow of 24 patients with hematologic malignancies who received HLA-identical sibling peripheral blood stem cell grafts after conditioning with fludarabine and 2 Gy of total body irradiation. The T-cell chimerism of blood and bone marrow was tightly correlated. Complete donor chimerism was reached earlier in the granulocytes than in the T cells. Mixed T cell chimerism was common at the time of onset of acute graft-versus-host disease (aGVHD), and both CD4(+) and CD8(+) donor T-cell chimerism increased with the occurrence of aGVHD grades II to IV (P =.0002 and P =.019, respectively). The rate of disappearance of recipient CD8(+) T cells was faster in patients with aGVHD grades II to IV than in patients without clinically significant aGVHD (P =.016). This observation indicates a role of graft-versus-lymphohematopoietic tissue reactions in creating complete donor T-cell chimerism. A donor CD8(+) T cell count above the median on day +14 increased the risk of subsequent development of aGVHD grades II to IV (P =.003). PMID- 15111934 TI - Prevention of veno-occlusive disease with defibrotide after allogeneic stem cell transplantation. AB - Veno-occlusive disease (VOD) of the liver occurs in 10% to 50% of patients after allogeneic stem cell transplantation, ranging from mild reversible disease to severe disease, with a mortality rate almost always close to 100%. Recently, promising results in the treatment of established VOD with defibrotide were reported. Therefore, defibrotide may be used as a prophylactic regimen for hepatic VOD in stem cell transplantation for hematologic malignancies. Fifty-two successive patients who underwent transplantation between October 1999 and June 2002 received defibrotide prophylaxis intravenously from day -7 to day +20 after transplantation in addition to heparin and were compared with historical controls who underwent transplantation successively between February 1997 and September 1999. In the defibrotide group, the maximum total bilirubin levels and the number of patients with serum levels exceeding 50 micromol/L were significantly lower than in the control group (5 of 52 versus 18 of 52, respectively; P =.004). None of the 52 patients developed VOD (Baltimore criteria), and no side effects occurred. These results were significantly different (P =.001) from controls (10/52 [19%] with VOD, 3 of whom died of severe VOD). In addition, day 100 event free survival was significantly higher in the study group (P =.02), with a trend toward better day 100 overall survival (P =.07). These results suggest that defibrotide given in addition to heparin may be an efficient prophylaxis for VOD. PMID- 15111940 TI - [Current methods of rehabilitation of patients with vestibular disorders]. AB - Efficacy of pharmacological, physical treatments in monotherapy and in combination was compared in rehabilitation of 173 patients with various forms of vestibular disorders. High efficacy of physical methods especially with the use of biological feedback is shown. Factors influencing final results of the above rehabilitation are analysed. Wide use of physical rehabilitation is recommended. PMID- 15111941 TI - [The role of betaserk in combined treatment of neurosensory hypoacusis and vestibulopathy in childhood]. AB - Betaserk treatment was given to 78 children aged 6 to 14 years suffering from acute or progressive neurosensory hypoacusis (n=62) and vestibulopathy (n=16). The best results were achieved in vestibulopathy in older children. Stable improvement of hearing in the range 10--20 dB was achieved in 16 (30.7%) children, stable hearing stabilization was seen in 32 (61.4%) cases. Betaserk was well tolerated, had no side effects. Thus, betaserk is effective in the treatment of children with vestibulopathy, acute and progressive neurosensory hypoacusis. PMID- 15111942 TI - [The role of audiological methods of investigation in diagnosis and treatment of chemodectomas and hemangiomas of the middle ear]. AB - Diagnostic value of some audiological methods of investigation (pure-tone audiometry, tympanometry, noisimetry) has been determined on the basis of examination and treatment of 114 patients with chemodectoma and hemangioma of the middle ear. Tympanometric evidence directly correlated with primary location and dissemination of the tumor. Changes on tympanograms and noisimetries correlate with the results of the tests with pressure of the neck's vessels and elevation of airpressure in the external acoustic meatus. Audiological findings are comparable with those of CT and MRT of the temporal bones and visualization of the tumor in surgery. High significance of the above methods was proved. PMID- 15111943 TI - [Clinical and acoustic characteristics of tracheoesophageal bypass with prosthesis]. AB - The authors analyse different aspects of tracheoesophageal bypass with prosthesis, how to prolong operation of voice prostheses. It is shown that replacement of the prostheses is frequently necessary not because of wear but rather due to fungal or microfloral affection. Radiotherapy had no adverse effects on prosthesis. Complications of tracheoesophageal bypass with prosthesis, acoustic characteristics of the repaired voice are described. PMID- 15111944 TI - [Gas composition of capillary blood of patients with laryngeal cancer in the course of conservative surgical treatment with polymeric endoprosthesis]. AB - Gas composition of capillary blood was studied in 120 patients with laryngeal cancer, primarily of stage IIIa (T3N0M0). Hypoxia was present in all the examinees whether or not laryngostenosis was clinically apparent. The analysis has shown that conservative surgery of laryngeal cancer with application of tubal polymeric endoprostheses corrects ventilatory disorders and improves blood gas composition. The degree of correction of ventilatory disorders depends on the kind of respiratory intraoperative maintenance. Gas composition appeared significantly better in the group of patients exposed to transtracheal high frequency artificial pulmonary ventilation than in the group exposed to volumic artificial pulmonary ventilation. PMID- 15111945 TI - [Application of endostents in tumor and scarry stenoses and main bronchi]. AB - To evaluate advantages and shortcomings of Dumon's, T-shape and polyflex endotracheal stents in patients with tumor and scarry stenoses of the trachea and main bronchi, we analysed the results of stenting trachea and main bronchi in 180 patients for 11 years (1992-2002). We observed the highest rate of granulations and dislocations for polyflex stents in the lowest adhesion of the sputum to their walls. Long-term results of stenting in scarry tracheostenosis demonstrate a moderate efficacy of T-stents and low efficacy of Dumon's stents. Our final conclusion is that a key method of treatment of tracheal scarry stenoses is a circulatory resection of the stenosed zone. Tracheal stenting remains important in urgent necessity of the treatment of decompensated stenoses, in preparation for radical operation and in contraindications to the latter. PMID- 15111946 TI - [Immunity in children with aquired cicatricial stenoses of the larynx and cervical trachea. Communication 3. Condition of phagocyting immune system]. AB - Macrophagal-phagocyting immunity was studied in 59 children with cicatricial acquired stenosis of the larynx and cervical trachea aged 1 year 9 months to 14 years 8 months. It was found that children with acquired cicatricial stenosis had a sharp depression of all functions of the cells of the macrophagal-phagocyting system (chemotaxis, absorptive, digestive, oxidation-reduction) as well as reduction of their number. This is, on the one hand, a defense reaction of the organism to continuous antigenic stimulation; on the other hand, this marks feasibility of progressive immunocomplex and/or autoimmune processes. The most prominent alterations were seen in patients with concurrent affection of the CNS. PMID- 15111947 TI - [Tympanic thermometry in diagnosis of exudative otitis media in children]. AB - Tympanic and axillary temperatures were measured in 90 children (161 ears) with exudative otitis media (EOM) and 20 otologically healthy children (40 ears). It was discovered that in EOM patients the difference between tympanic and axillary temperatures is greater than in healthy children. This increased difference is caused by the state of the tympanic mucosa. The influence of surgical and conservative treatments on tympanic mucosa temperature and, therefore, on the difference between tympanic and axillary temperatures in these patients are outlined. It is recommended to use tympanic thermometry with consideration of case history data and otoscopic picture as a screening method for diagnosis of EOM in mass prevention examinations of children, in family physician practice and in selection of children for more comprehensive check-up in diagnostic centers as well as for control over treatment results in EOM children. PMID- 15111948 TI - [Optimization of therapeutic and diagnostic processes in the system of obligatory medical insurance in an outpatient ENT department]. AB - How to organize effective activity of an outpatient ENT department in the system of obligatory medical insurance (OMI) is considered. To achieve this purpose, an analysis of everyday activity of an outpatient department was made with estimation of reference morbidity, epidemiological data on the residents of the Central Moscow District. Measures for introduction of novel technologies in therapeutic outpatient ENT practice are proposed. They include introduction of new otorhinolaryngological services, intensification of the work of ENT specialists, stimulation of their interest in mastering allied specialties, stimulation of medical assistants interest in mastering new procedures, estimation of ENT services cost and minimizing the difference between the cost of the service and salaries of the medical personnel. PMID- 15111949 TI - [Surgical treatment of patients with odontogenic maxillary sinusitis]. AB - Outcomes of surgical treatment of maxillary sinusitis of odontogenic origin have been studied in 31 patients. A glue composition MK-9M was applied. This procedure can be recommended as a method of choice in management of perforative maxillary sinusitis. PMID- 15111950 TI - [Postobturation pain associated with endodontic treatment]. AB - Postobturation pain of different origin after endodontic treatment was studied in 100 patients after treatment of 151 teeth with filling of 264 channels. 34.4% patients complained of pain. The incidence and intensity of pain were scored. Postobturation pain clearly depended on the number of treatment sessions, number of channels filled in one tooth, and excessive filling "beyond the apex". The method of "crown down" preparation using completely rotating profile instruments and GT rotary files proved to be most effective as regards prevention of postoperative pain. PMID- 15111951 TI - [Problems of examinations and diagnosis of periodontal diseases]. AB - More then 50% of the examined periodontal patients are usually qualified as the ones, suffering from the periodontitis of severe or half-severe form of disease. The analysis of questionnaires and medical histories revealed the inconvenience in the investigation methods and diagnostics of the periodontal patients. There is accentuated the need of curly diagnostics and complex treatment. PMID- 15111952 TI - [A method for treatment of jaw fractures using temporary intraosseous implants]. AB - The authors suggest a new method for treatment of mandibular fractures. This method consists in insertion of implants (19 mm long, 1.8 mm thick) made of medical stainless steel into the bone, repositioning of fragments, and subsequent fixation by intermaxillary traction. The study showed that this method is more physiological, causes less side effects, and requires shorter hospital stay in comparison with the traditional treatment. PMID- 15111953 TI - [Social and organizational problems and approaches to optimization of diagnosis of the buccal mucosa carcinomas]. AB - The incidence of malignant tumors of the buccal mucosa and stages of carcinoma were analyzed in 565 primary patients for a period of 10 years. The study showed social and organization problems in the disease diagnosis: refusal of patients from medical consultation, unjustified omitting of examination of the oral cavity aimed at cancer detection during preventive examinations of the population and treatment of odontogenic diseases, erroneous interpretation of pathological changes in the buccal mucosa, unjustified long observation and treatment by non oncological specialists, incompetence of physicians, etc. Approaches to optimization of diagnosis are outlined: improvement of the efficiency of preventive examinations by non-oncological specialists, specialized upgrading of dentists and increase of their professional responsibility, and early use of morphological methods of examination of pathological processes in the buccal mucosa. PMID- 15111954 TI - [Reconstructive operations in the treatment of carcinoma of the lower lip]. AB - The authors sum up the results of combined and reconstructive surgical treatment of 213 cases with squamous-cell carcinoma of the lower lip, carried out in Tver Oncological Center during the recent decade. Oncological and functional esthetic results of different methods of primary cheiloplasty are analyzed. The findings confirm that clinoid resection of the lip is not permissible as a method of treatment and that indications for resection of the lower lip without primary cheiloplasty should be limited. The authors suggest their own method for surgical repair of the lower lip, which was used in the treatment of 23 patients with carcinomas. Evaluation of the adequacy and efficiency of the method and analysis of the incidence and type of postoperative complications, function of the lower lip after its repair, and esthetic results showed high esthetic and functional efficiency of the new method, due to lesser trauma of the soft tissues, lesser blood loss, and shorter time required for elevation of triangular mucous flaps, adequate blood supply to triangular mucous grafts and lower risk of their aseptic necrosis, anatomic topographic division of vertical fragments of skin and mucous wounds of the lip, and lower risk of secondary wound infection. PMID- 15111955 TI - [Role of ligation of the external carotid artery in the treatment of malignant tumors of the tongue]. AB - This paper presents analysis of the efficiency of ligation of the external carotid arteries in the treatment of malignant tumors of the tongue, based on the data of Department of Head and Neck Diseases of Tver Regional Clinical Oncological Center for two recent decades. A total of 87 case histories of patients in whom 26 ligations of the external carotid artery were performed, are analyzed. This operation is used if the tumor is located in the posterior third and root of the tongue and if it involves the tissues of the bottom of the oral cavity. If the tumor was located in the two anterior thirds of the tongue or a small tumor is located at the root of the tongue, this intervention was not used. It is indicated for decreasing the hemorrhage during surgery for malignant tumors of the tongue and for preventing postoperative hemorrhages, hemorrhages from degrading tumor in inoperable patients, in patients with threatened hemorrhage before radiotherapy, and during bleeding. PMID- 15111956 TI - [Permanent adhesive dentures]. AB - The longitudinal clinical study (up to 15 years of clinical performance) was carried out to evaluate the efficiency of the different resin-bonded prostheses - fixed partial dentures, veneers, splints. PMID- 15111957 TI - [Clinical and anatomic features of the denture bed in a toothless mandible and orthodontic strategy]. AB - Clinical anatomical features and orthodontic strategy were studied in a specially selected group of 72 patients aged 56-86 years with completely lost teeth and unfavorable clinical conditions on the toothless mandible. The authors claim that their strategy and method of treatment of this patient population are effective and recommend it for wide practice. PMID- 15111958 TI - [Clinical picture and orthodontic treatment of adult patients with cleft lip and palate]. AB - There are represented the results of clinical investigation of 15 patients in age from 15-54 years with cleft lip and palate. There is given the detailed clinical description and the peculiarities of prosthetic rehabilitation of those patients. PMID- 15111959 TI - [Acute thrombophlebitis in the territory of saphena magna vein]. AB - Problems of strategy and policy of surgical treatment of acute thrombophlebitis in the territory of saphena magna vein are discussed. Results of urgent radical combined venectomy in 76 patients are presented. A gas technique of subfascial dissection of perforant veins (SEPS) was used in 52 patients for the first time both for diagnosis and adequate treatment of thrombosed perforants. Spread of thrombosis from superficial veins to perforants was revealed in 14.3% patients with C3-C5 class of venous dysfunction (CEAP classification). Isolated and local forms of crus varicothrombophlebitis that must be urgently operated are distinguished. The mean hospital stay was 8.7 bed-days. PMID- 15111960 TI - [Prognosis of complications after reconstructive surgeries on the aorto-iliac segment]. AB - Evaluation of efficacy of neuronet technologies for prognosis of specific complications after reconstructive surgeries on aorto-iliac segment was made. A learning sample consisted of 500 reconstructive surgeries, a test one - 161. High efficacy of analog neuron nets was revealed. Prognosis of early thrombosis, postoperative bleeding and also of lethal outcome demonstrated better results, mistake of prognosis was 3.7, 1.2 and 4.6%, respectively. Prognosis of infectious complications and long-term outcomes demonstrated worse results, mistake of prognosis was 7.0 and 10.5%, respectively. PMID- 15111961 TI - [Purulent-septic complications after reconstructive surgeries in the patients with chronic lower limb ischemia]. AB - Purulent-septic complications after reconstructive surgeries in patients with chronic lower limb ischemia are the great problem of vascular surgery. Analysis of 135 reconstructive surgeries in chronic lower limb ischemia was carried out. Purulent and septic complications were reported in 17 (12.59%) patients. Variants of treatment and outcomes are described. PMID- 15111962 TI - [Correlation of extremity safety and method of reconstructive surgery on femoral popliteal segment]. AB - Long-term results of 660 reconstructive surgeries on the arteries of femoral popliteal segment have been analyzed. In critical limb ischemia surgeries with PTFE grafts are most effective - in 5 years 77.1% of extremities were saved compared with 69.6% after autovenous reconstructions. Use of gelatinized velour dacron (GVD) grafts (when PTFE or autovenous grafts are absent) permits to save 62.4% of extremities in 5 years. Endarterectomies (EAE) and bypass surgeries with fluorolavsan (FL) grafts should not be regarded as surgeries of choice in critical limb ischemia. Safety of limbs after surgeries with PTFE, GVD and autovenous grafts performed in the stage of claudication does not differ significantly. Cumulative safety of extremities after EAE and bypass with FL grafts is worse but high. In the first month after surgery 29.3% of all amputations were performed, in the first six months - 47.8%. PMID- 15111963 TI - [Diagnosis, treatment and prophylaxis of postoperative recurrent nodular goiter]. AB - Results of surgical treatment of 81 patients with recurrent goiter and 79 patients with benign nodular forms of goiter are analyzed. Based on clinical data, results of ultrasonic and histological examinations, it was determined that recurrence of goiter does not depend on volume of surgery. It is seen in patients with subclinical forms of postoperative hypothyreosis and in half of cases with lymphoid infiltration of I - IV degree. Dispensary observation and timely prescription of suppressive therapy, if it is necessary, permit to reduce number of goiter recurrences from 8.2 to 2.5%. PMID- 15111964 TI - [Wound-adapter in the treatment of purulent wounds]. AB - The method of closure of soft tissues purulent wounds with wound-adapter of original construction has been developed and used in clinical practice. This method provides complete reposition and adaptation of wound margins without suture eruption. Good results (primary closure) were achieved in 85.8% cases in the study group, and in 49.7% cases in the control group, satisfactory results (secondary closure partly) - in 14.2 and 50.3% cases, respectively. Hospital stay was reduced from 27.6 +/- 1.5 to 22.4 +/- 0.7 bed-days (p < 0,05). PMID- 15111965 TI - [Analysis of causes of unsatisfactory results of rectocele surgical treatment and ways of their correction]. AB - Causes of recurrences and unsatisfactory results of rectocele surgical treatment (except wound suppuration after surgery) were analyzed. It was concluded that prolapse of rectal mucosa and (or) severe muscular atrophy of the pelvic fundus uncorrected during transvaginal correction of rectocele are the main causes. It is recommended to resect mucosal excess by a traditional or A. Longo's method in the former case and to use synthetic net in the latter. This brings about favorable results in these patients. PMID- 15111966 TI - [Choice of surgical method in rectal cancer]. AB - Based on randomized study of 589 patients radically operated for rectal cancer, rational surgical policy was developed. Efficacy of rectal resection depending on the approach, scope of mesorectumectomy and anal demucosation was studied, comparison of purulent complications number, sphincter function and early recurrence in 315 patients was carried out. It is demonstrated that methods of rectal resections with lowest per cent of purulent complications lead to the worst functional results and increase number of local recurrences. Differential approach to choice of resection or extirpation method is proposed, clear indications are formulated. PMID- 15111967 TI - [Dynamics of reparative processes in intestinal wound during formation of different entero-enteroanastomoses]. AB - Suture insufficiency is the most severe complication in gastrointestinal surgery. Efficacy of various intestinal anastomoses and main processes in zones of intestinal anastomosis in different terms after surgery were studied experimentally. Qualitative and quantitative assessment of morphofunctional changes of tissues in anastomotic help to formulate significant criteria for choice of methods of intestinal anastomosis formation and reduce number of postoperative complications. PMID- 15111968 TI - [Register of Acute Coronary Syndromes in Russia: Management and Inhospital Outcomes in ST Elevation Acute Coronary Syndrome]. AB - Data of about 50 consecutive patients with acute coronary syndromes aged at least 18 years admitted to 59 hospitals in different Russian cities were collected from November 2000 to July 2001. In 1412 patients included into registry presumably ischemic symptoms within previous 24 hours were associated with ST-segment elevation or left bundle branch block on ECG. Demographics, history, characteristics of acute coronary syndrome, management and outcomes during hospitalization as well as diagnoses at presentation and discharge were analyzed. Markers of myocardial necrosis were measured in 61% of patients. Rates of interventions that may improve outcome appeared to be low. In acute phase aspirin was used in 79% (contraindications were reported in 6.2%). Within 12 hours of symptoms onset thrombolysis was performed in 12.9%, coronary angioplasty in 1.2%. In patients hospitalized within 12 hours of symptoms onset these rates were 21.3 and 1.9%, respectively. Beta-blockers were prescribed no more than in 60% of cases (fist dose intravenously in 4.3%). Lipid lowering drugs were recommended at discharge to 12.3% of patients (to 21.1% of those with known hypercholesterolemia). ACE inhibitors during hospitalization were used in 68.1% of patients. After acute phase of the disease coronary angioplasty was performed in 5 patients, CABG in 1. However hospital mortality appeared to be not high (8.5% in general, 10.1% in patients with overt acute myocardial infarction at presentation). Reinfaction rate in this registry was impossible to assess, angina recurrences were registered in every fifth patient. PMID- 15111969 TI - [Results of nonmedical interventions in multicenter randomized open study of efficacy of lifestyle modification and therapy with quinapril in patients with obesity and hypertension]. AB - There is a pathogenetic interrelationship between obesity and hypertension. Moreover it has become evident that the use of most modern and active antihypertensive drugs can not be effective without concomitant treatment of obesity. This study was conducted in 18 cities of Russian Federation in order to demonstrate influences of lifestyle changes on hypertension and levels of cardiovascular risk factors. The objective was to attract attention of physicians and patients to the problem of obesity and hypertension and to their inherent dangers. PMID- 15111970 TI - [Rheological properties of blood and endothelial function in patients with hypertensive disease]. AB - Patients with hypertension are characterized by elevated platelet and erythrocytes aggregation and presence of relationship between rigidity of red blood cells and myocardial mass index. They also have increased number of leukocytes in peripheral blood, enhanced adhesive ability of neutrophils, increased representation of integrin receptors on lymphocytes and monocytes, and expression of Fas-receptors. These features evidence for augmented functional activity of leukocytes in hypertensive disease. Changes of rheological parameters of blood occur before impairment of vasomotor endothelial function. Augmentation of viscous properties of blood and functional activity of leukocytes aggravates hemodynamic disturbances in hypertension and can facilitate myocardial and vascular remodeling. PMID- 15111971 TI - [Complex assessment of vasomotor function of vascular endothelium in patients with hypertension]. AB - Vasomotor function of vascular endothelium was studied in 62 patients with grade 1-2 hypertension with moderate and high added risk. Methods included study of brachial and middle cerebral artery endothelium dependent and independent vasodilation/vasoconstriction, measurement of plasma levels of nitric oxide metabolites (NO(n)-), endothelin-1, and antithrombin, as well as registration of their changes during vasomotor tests with calculation of integral indexes. Most patients with hypertension differed from controls by preponderance of vasoconstrictor over vasodilator reactions both in peripheral and cerebral vascular bed. At the same time patients with hypertension had pronounced dissociation between vasomotor responses of cerebral and peripheral vessels compared with subjects with normal blood pressure (p<0.05). Besides lowered basal level of NO(n)- and high concentration of endothelin-1 patients with hypertension were characterized by hyperreactivity of nitricoxidergic system, augmented lability of endothelin producing system, and impaired athrombogenecity of vascular endothelium. Complex assessment of vasomotor function of vascular endothelium by sequential vasoactive tests characterizes functional and metabolic activity of cerebral and peripheral vessels and can be used for improvement of risk stratification and monitoring of efficacy of treatment of patients with hypertension. PMID- 15111972 TI - [The state of microcirculation during use of long acting metoprolol]. AB - The aim of the study was to assess the effect of prolonged action metoprolol on the microcirculation in patients with essential hypertension (EH). We studied 51 patients with EH aged 53,4+/-4,5 years. After 7-10 days of "wash-out" period and after 8 weeks of prolonged action metoprolol (betaloc ZOK) the state of microcirculation was investigated with the use of laser Doppler flux-metry (LDF). We found significant decrease in basically enlarged basal blood flow and the level of sympathetic activity, increase in the amplitude of cardiodependent waves, normalization of basal blood flow decrement in postural test, increase in capillary blood flow reserve in patients with stage I-II EH. We failed to observe any significant changes in the microcirculatory parameters in patients with stage III EH. Treatment with prolonged action metoprolol led to increase in the number of patients with normocirculatory type of microcirculation primarily due to the decrease in the prevalence of hyperemic type. We conclude that prolonged action metoprolol does not alter microcirculation in patients with EF. Moreover, at the early stages of EH it improves the functional state of peripheral hemodynamics due to suppression of sympathetic activity. PMID- 15111973 TI - [Clinical, angiographical, and procedural causes of acute vessel closure during transluminal coronary intervention]. AB - AIM: To elucidate factors related to acute vessel closure (AVC) after transluminal coronary intervention. METHODS: From population of 10439 patients subjected to transluminal coronary intervention 2 groups were formed: with (n=885) and without (n=885) acute vessel closure (AVC). Twenty five clinical, angiographical and procedural characteristics of patients of these 2 groups were included into mono and multifactorial logistic regression analysis. RESULTS AND CONCLUSION: The following factors were univariate predictors of acute vessel closure: smoking [odds ratio (OR) 1.42], unstable angina (OR=2.130, acute myocardial infarction within previous 24 hours (OR 2.76), cardiogenic shock (OR 4.31), urgent procedure (OR 1.94), eccentric stenosis (OR 1.67), calcified lesion (OR 2.21), preexisting thrombosis (OR 3.79), lacerated complicated stenosis (OR 2.02), tortuous lesion (OR 1.35), low operator experience (OR 3.37), balloon angioplasty as sole procedure (OR 1.66), concomitant rheolytic thrombectomy (OR 1.95), urgent stenting (OR 1.45). Elective stenting significantly lowered risk of acute vessel thrombosis. Multifactorial step-up analysis selected the following independent predictors of AVC: smoking, acute myocardial infarction within previous 24 hours, cardiogenic shock, preexisting thrombosis, lacerated complicated stenosis, and concomitant rheolytic thrombus extraction. Thus only elective stenting significantly reduced risk of AVC. PMID- 15111974 TI - [Effect of trimetazidine on regressione of ischemic dysfunction of myocardium after coronary bypass surgery on beating heart]. AB - AIM: To study effect of trimetazidine on restoration of hibernating myocardium after myocardial revascularization on beating heart. MATERIAL: Patients with ischemic heart disease subjected to direct myocardial revascularization on beating heart: 25 patients received trimetazidine (60 mg/day) in pre and postoperative periods and 30 patients did not. METHODS: Echocardiography, veloergometry, 6 minute walk test, myocardial scintigraphy with Tl-199. RESULTS: Course (35 days) treatment with trimetazidine provided significant decrease of frequency and severity of episodes of angina, reduction of nitrate consumption, enhancement of tolerance to physical exercise, improvement of myocardial perfusion manifested as significant decrease of mean size of transient perfusion defects. CONCLUSION: These results demonstrate efficacy and expediency of trimetazidine use in therapy of reversible myocardial dysfunction in patients with ischemic heart disease subjected to direct myocardial revascularization. PMID- 15111975 TI - [Hemorheological changes during mammary coronary bypass grafting]. AB - Hemorheological studies were performed in 44 patients subjected to mammary coronary bypass surgery during induction of anesthesia, at initiation of cardiopulmonary bypass, at the end of operation, on the 1(st) and 2(nd) or 3(rd) postoperative day. Methods included determination of whole blood and plasma viscosity with estimation of suspension stability and calculation of Caisson viscosity and fluidity limit. Rigidometry was used for assessment times of linear (T(1)) and three dimensional (T(2)) aggregates formation, final aggregates dimensions, total hydrodynamic strength of aggregates (beta), index of strength of especially large aggregates. Indexes of aggregation and deformation of erythrocytes were also calculated. Cardiopulmonary bypass was associated with lengthening of T(1) and shortening of T(2), increase of aggregates dimension and their hydrodynamic strength beta and lowering deformability of erythrocytes. By the end of operation increase of difference between Caisson and asymptotic viscosity was noted. Postoperative period was characterized by improvement of deformability of erythrocytes, prolongation of T(2), increases of aggregates dimension, their hydrodynamic strength beta, and fluidity limit. Thus measures of intensive therapy should be directed at correction of disturbances of rheological properties of blood with consideration of their changes specific for cardiosurgical patients. PMID- 15111976 TI - [Assessment of phenomenology of attacks of atrial fibrillation before and after treatment with clonasepam]. AB - AIM: To assess phenomenology of attacks of atrial fibrillation using psychovegetative index (PVI) and to compare PVI values obtained before and after treatment with clonasepam in patients with paroxysmal atrial fibrillation. MATERIAL: Patients with nonrheumatic atrial fibrillation (n=105). METHODS: PVI was calculated as ratio of difference between sums (numbers) of cardiac and noncardiac symptoms to sum (number) of cardiac symptoms. RESULTS: Psychovegetative disturbances predominated during attacks of atrial fibrillation and because of that PVI was less than zero. Greatest values of PVI were noted during attacks of idiopathic atrial fibrillation, in women, in patients with high initial level of personal anxiety, in patients with frequent and nocturnal attacks. CONCLUSION: Patients with negative or zero PVI require drugs for correction of psychovegetative disturbances. Therefore it is expedient to use PVI in patients with attacks of atrial fibrillation to select candidates for psychotropic therapy and to evaluate its efficacy. PMID- 15111977 TI - [Aspartic Acid and mannitol enhance protective efficiency of asanquineous cardioplegic solution]. AB - The aim of this work was to assess effects of a novel asanquineous cardioplegic solution (CP-5), buffered with trisamine (pH 7.6+/-0.1 at 22 degrees C) and containing 21.5 mM aspartic acid and 20.0 mM mannitol, on postischemic functional and metabolic recovery of isolated rat heart. A modified Ringer solution with 25 mM KCl (pH 7.6+/-0.1 at 22 degrees C) and the St. Thomas' cardioplegic solution (pH 7.8+/-0.1 at 22 degrees C) were used as controls. Osmolarity of all cardioplegic solutions were 340+/-5. After 20-min initial perfusion according to Neely (steady state) the hearts were subjected to 40-min normothermal total ischemia followed by 30-min antegrade reperfusion. Cardioplegic solutions were infused prior to ischemia at rate of the initial coronary flow for 5 min at room temperature. During reperfusion the hearts of CP-5 group completely recovered coronary flow and significantly enhanced restoration of the majority functional indices compared to the hearts in both control groups. This effect was combined with less lactate accumulation and preservation of higher ATP and phosphocreatine (PCr) levels in the heart tissue by the end of ischemia and, probably was induced by inclusion of aspartic acid into composition of CP-5. By the end of reperfusion the hearts treated with CP-5 completely recovered PCr content and restored ATP level up to 65.2+/-4.6% of initial one. A better energy state of reperfused hearts in CP-5 group was accompanied by reduction of myocardial lactate tissue to the preischemic value. Restoration of ATP, PCr and lactate content was significantly poor in both control groups during reperfusion. The least formation of a spin adduct of the short life oxygen radicals was found in the myocardial effluent of the hearts of CP-5 group at the early reperfusion using EPR technique. These data suggest a reduced release of oxygen radical generating systems from postischemic myocardium into perfusate due to antioxidant effect of mannitol. The obtained results substantiate addition of aspartic acid and mannitol to the asanquineous cardioplegic solution, buffered with trisamine, to enhance efficacy of myocardial protection against ischemia and reperfusion injury. PMID- 15111978 TI - [Left Ventricular Hypertrophy in Hypertension. Part III. Reversibility of Left Ventricular Hypertrophy During Treatment With Antihypertensive Drugs]. AB - Results of prospective trials of effects of various antihypertensive drugs on left ventricular hypertrophy in patients with hypertensive disease are reviewed. According to 2 meta-analyses angiotensin converting enzyme inhibitors are most effective inducers of regression of left ventricular hypertrophy. However in comparative randomized trials in patients with hypertension ability of diuretics, lipophilic beta-adrenoblockers, long-acting calcium antagonists, and angiotensin receptor blockers to cause regression of left ventricular hypertrophy was not inferior to that of angiotensin converting enzyme inhibitors. PMID- 15111979 TI - [Arterial hypertension in a framework of metabolic syndrome: special features and principles of drug correction]. AB - About 60% of patients with mild and moderate hypertension have insulin resistance and half of them have clinically manifest metabolic syndrome which comprises abdominal obesity, hyperlipidemia, impaired glucose tolerance, hypertension and insulin resistance. In a framework of metabolic syndrome hypertension is characterized by disturbed circadian profile without nocturnal blood pressure lowering and concentric left ventricular hypertrophy. There exist 2 mechanisms of linkage between hypertension and metabolic syndrome: impaired ion transport and neurohormonal and humoral activation. Antihypertensive drugs for correction of hypertension in metabolic syndrome should be long acting, provide protection of target organs, and induce positive or neutral metabolic effect. Together with normalization of blood pressure these actions can cause lowering of risk of atherosclerosis development. Representatives of the following classes of antihypertensive agents can be used as drugs of choice: angiotensin converting enzyme inhibitors, long-acting calcium antagonists, selective beta1 adrenoblockers, and thiazide diuretics. PMID- 15111980 TI - A prospective randomized comparison of extensive prostate biopsy to standard biopsy with assessment of diagnostic yield, biopsy pain and morbidity. AB - In a prospective randomized study, we compare standard prostate biopsy to extensive biopsy utilizing intravenous conscious sedation (IVCS). Initial biopsy patients (n=197) were randomized to either standard biopsy using intrarectal lidocaine gel (6-12 biopsies, mean 10.1) or extensive biopsy (24 biopsies) using IVCS. Cancer detection and urinary symptoms were no different between groups. However, biopsy pain was rated significantly lower and satisfaction significantly higher in the extensive biopsy group. Temporary urinary retention occurred in 4% of the extensive biopsy group. Extended biopsy with 24 samples does not improve cancer detection compared to standard biopsy when 10 cores are obtained. Extensive biopsy is very well tolerated and associated with less pain and more satisfaction than standard biopsy. PMID- 15111981 TI - A 3-y prospective study of health-related and disease-specific quality of life in patients with nonmetastatic prostate cancer treated with radical prostatectomy or external beam radiotherapy. AB - We assessed the longitudinal alteration of the quality of life (QOL) of patients with localized prostate cancer after radical prostatectomy or hormonoradiotherapy during 3-y follow-up. In addition, we examined the impact on QOL of initiation of second treatment after failure of primary treatment. In all, 135 patients with localized prostate cancer who underwent radical retropubic prostatectomy (RP) (N=84) or external beam radiotherapy with neoadjuvant hormone (XRT) (N=51) at our institute and who had a minimum follow-up of 3 y were included in this study. Data were collected prospectively, at baseline, at 3 months after treatment, at 1 y, and annually thereafter. QOL, generic and disease-targeted was evaluated using the European Organization for Research and Treatment of Cancer Prostate Cancer QOL Questionnaire, the Sapporo Medical University Sexual Function Questionnaire, the International Prostate Symptom Index Quality of Life Score and similar questions regarding bowel function. Repeated-measures ANOVA revealed significantly different patterns of alteration in the domains of QOL, with the exception of several domains, between the RP and XRT groups. Rapid decline of sexual function and increase in sexual bothersomeness were followed by slight amelioration throughout follow-up in the RP group, and did not change thereafter in the XRT group. Overall satisfaction with urinary condition significantly improved after treatment and that with bowel condition was stable during follow up in both of the groups. Failure of primary treatment and initiation of salvage treatment had no impact on QOL. This prospective study revealed longitudinal alteration of QOL status of patients undergoing treatment for localized prostate cancer, but did not yield any conclusions regarding effect of treatment failure and second treatment on QOL due to small sample size. It should be noted that different instruments for assessment of QOL can generate different outcomes. PMID- 15111982 TI - Thimerosal and autism. PMID- 15111983 TI - Resistance to the orexigenic effect of ghrelin in dietary-induced obesity in mice: reversal upon weight loss. AB - BACKGROUND: Ghrelin, an endogenous ligand for growth hormone secretagogue receptor (GHS-R), is known to increase food intake in lean humans and rodents. In addition, ghrelin levels are increased by fasting in lean rodents and are elevated before meals in humans, suggesting an important role for ghrelin in meal initiation. However, in obese human, circulating ghrelin levels were found to be significantly reduced as compared to lean individuals. OBJECTIVES: To evaluate whether circulating ghrelin levels, as well as ghrelin sensitivity, are decreased in obese individuals in order to limit its effect on food intake. DESIGN: : Lean C57BL/6J mice fed a chow, a low- (LFD) or a high-fat diet (HFD) were used to determine ghrelin regulation and secretion as well as ghrelin sensitivity. MEASUREMENTS: Plasma ghrelin levels were measured in low- and high-fat fed mice. Ghrelin-induced food intake was measured in chow, low- and high-fat fed mice. RESULTS: We measured ghrelin levels in lean and diet-induced obese mice, fed on an LFD or an HFD, respectively. We observed that not only ghrelin secretion was reduced in obese mice but its diurnal regulation was also lost. In addition, we failed to observe any change in ghrelin secretion upon fasting and refeeding. Moreover, we observed that the sensitivity to the orexigenic effects of exogenous ghrelin was reduced in obese mice when compared to lean mice fed a chow or a LFD. The insensitivity of obese mice to ghrelin was improved upon weigh loss. CONCLUSION: : Altogether, these results indicate that ghrelin secretion and regulation is impaired in dietary-induced obesity in mice and suggest that ghrelin inhibition could prevent weight regain after weight loss. PMID- 15111984 TI - Response of the alternative complement pathway to an oral fat load in first degree relatives of subjects with type II diabetes. AB - OBJECTIVE: To investigate levels of components of the alternative pathway of complement, and of two activation products, ASP and Bb, in persons ranging in insulin resistance both fasting and following the consumption of a high-fat, low carbohydrate meal. SUBJECTS: Healthy controls (n = 17) and normoglycaemic first degree relatives of patients with type II diabetes (n = 15). MEASUREMENTS: All subjects had normal glucose tolerance. Blood was collected for the measurement of plasma glucose, insulin, triglycerides and free fatty acids. Body composition was assessed with dual energy X-ray absorptiometry (DEXA) and whole-body insulin sensitivity with a euglycaemic, hyperinsulinaemic clamp. Basal and postprandial values over 6 h were determined for plasma C3, B, D, Bb and ASP. Basal levels of C1q, C4 and CRP were also determined. RESULTS: Controls did not differ significantly from the relatives of patients with type II diabetes for any metabolic parameter except in their degree of insulin resistance and central fat (kg). Across all subjects, basal levels of C3, but no other complement protein, were correlated with insulin resistance. Native complement proteins, but not ASP or Bb, were correlated with body mass index and the amount (kg) of central fat. Basal levels of C3 and factor B were significantly higher in the relatives group, whereas factor D and the classical pathway proteins C1q and C4 did not differ between the two groups. Postprandially, levels of factor D were significantly reduced in both groups. ASP levels also fell postprandially, the decline achieving significance in the relatives group. CONCLUSIONS: Elevated levels of C3 and factor B in the diabetic relatives group may have resulted from increased synthesis by adipose tissue. There was no evidence of alternative pathway activation in response to a fat meal in terms of ASP or Bb production, or significant consumption of C3 and factor B. These data do not support an essential requirement of the hypothesis that ASP is produced in response to the intake of fat. PMID- 15111985 TI - Apolipoprotein B signal peptide polymorphism and plasma LDL-cholesterol response to low-calorie diet. AB - OBJECTIVE: To assess the effect of the apolipoprotein B Ins/Del signal peptide. Polymorphism on plasma lipid levels in overweight subjects before and after a low calorie diet. DESIGN: Diet intervention study (25% reduction in energy intake during 2.5 months) in relation to genetic factors. SUBJECTS: A total of 231 unrelated patients (146 women/85 men) recruited on the basis of body mass index (BMI)> or =25 kg/m(2). MEASUREMENTS: BMI, waist to hip ratio, blood lipids and lipoproteins, at entry and after 2.5 months, determination of apo B Ins/Del genotypes. RESULTS: On spontaneous diet, subjects carrying the Del allele had higher LDL-cholesterol (Del/Del: 3.97+/-0.62 mmol/l; Ins/Del: 3.87+/-1.01 mmol/l; Ins/Ins: 3.61+/-0.88 mmol/l) (P=0.038). When submitted to low-calorie diet, subjects with Del/Del genotypes reduced their LDL-cholesterol (-16.8%) more than subjects with Ins/Del or Ins/Ins (-4.7% and +0.9%, respectively) (P=0.001). CONCLUSION: In overweight or obese people, the response of plasma LDL-cholesterol levels to low-calorie diet is modulated by genetic variation at the apo B locus. Overweight subjects with the Del allele of the apo B signal peptide polymorphism are predisposed to high LDL cholesterol levels but their LDL cholesterol responds well to diet. These results demonstrate the importance of the interaction between genes and nutritional environment in the determination of the lipid levels. PMID- 15111986 TI - A Pavlovian approach to the problem of obesity. AB - During the past 15-20 y, the incidence of overweight and obesity in the United States has grown rapidly. The processes that underlie this alarming trend remain largely unspecified. We hypothesize that degradation of the ability to use certain orosensory cues to predict the caloric consequences of intake may contribute to overeating and excessive weight gain. The results of two preliminary studies with rats are consistent with this hypothesis. In one study, the ability of rat pups to regulate their caloric intake after consuming a novel high-calorie, sweet food was disrupted if they had received prior training with sweet tastes that failed to predict the caloric consequences of eating. Another study found that altering the normal predictive relationship between food viscosity and calories led to increased body weight in adult rats. Dietary factors that degrade the relationship between sweet tastes, food viscosity and calories may contribute to overeating and weight gain. PMID- 15111987 TI - The pharmacogenomics of selective serotonin reuptake inhibitors. AB - The introduction of selective serotonin (5-HT) reuptake inhibitors (SSRIs) has significantly improved the pharmacological treatment of a range of psychiatric disorders. Nevertheless, despite the undoubted advantages of antidepressant treatment in terms of improved tolerability to therapy while maintaining a high level of efficacy, not all patients benefit from it; an appreciable proportion do not respond adequately, while others may show adverse reactions. The necessary change of the initial treatment choice often requires extended periods for the remission of symptomatology. Such difficulties could be avoided if it should be possible to determine more quickly the most suitable drug. Several factors have been thought to influence the outcome of antidepressant therapy. Among the factors influencing the interindividual variability in response to treatment with SSRI, differences in genetic features may play a significant role. Several genetic polymorphisms have been associated with therapeutic SSRI response, including genetic variants of the 5-HT transporter, 5-HT-2A-receptor, tryptophan hydroxylase, brain-derived neurotrophic factor, G-protein beta3 subunit, interleukin-1beta and angiotensin-converting enzyme, although with conflicting results; also cytochrome P450 drug-metabolising enzymes may bear a particular importance, although further corroboration of the findings is necessary, and further key participating genes remain to be identified. The hope is that the identification of these genetic components will eventually facilitate the development of a customised SSRI treatment. PMID- 15111988 TI - Variable drug metabolism genes in Arab population. AB - Cataloging interethnic differences in the distribution of genotypes of drug metabolic genes provides valuable information for profiling the pharmacogenetics of a population. We used PCR analysis to catalog the frequencies of alleles and genotypes for CYP1A1, NAT2, GSTs, MTHFR, MTR (MS) and NQO*1 in Arabs. The frequencies of alleles and/or genotypes for CYP1A1*2A, GSTT1 null, GSTT1 and GSTM1 double null, and GSTP1 A1578G in Arabs were significantly higher than those reported in Caucasians. However, the distribution of NAT2 acetylator phenotypes in both populations was similar. In contrast, the frequencies of MTHFR 677T allele and the combined (677+1298) genotypes for low activity were lower than those reported in Caucasians. Other alleles in Arabs, including CYP1A1 T3801C and GSTP1 A1578G were present in frequencies similar to Africans. The overall profile of variations in metabolism genes in Arabs is thus unique. PMID- 15111989 TI - Further evidence for a possible association between serotonin transporter gene and lithium prophylaxis in mood disorders. AB - We previously reported an association between the functional polymorphism in the upstream regulatory region of the serotonin transporter gene (SERTPR) and the prophylactic efficacy of lithium in a sample of 201 Italian subjects affected by Mood disorders. The aim of the present study was to replicate analyses on an independent sample. In total, 83 subjects affected by Bipolar disorder were recruited in the Mood Disorders Clinic of the Eginition Hospital of the Athens University, Medical School Department of Psychiatry. All patients were administered with lithium as prophylactic therapy and they were prospectively observed for at least 3 years. Subjects were typed for their SERTPR variant using polymerase chain reaction techniques. SERTPR variants were associated with lithium outcome among those subjects who had few manic episodes before lithium treatment and, as a trend, among subjects who received a high daily dose of lithium (> or =1200 mg/die). In both cases, subjects with the l/l variant showed a higher probability to develop an illness episode within 3 years of prophylactic treatment with lithium. The present study confirmed our previous observation of a better response of SERTPR*l/s carriers, but could not confirm a poor efficacy in subjects with the SERTPR*s/s genotype. Notwithstanding the conflicting results, SERTPR variants are a possible liability factor for lithium long-term efficacy in mood disorders. Further studies on independent and large samples are required to determine the reliability and direction of the possible association between SERTPR variants and lithium outcome. PMID- 15111990 TI - A case of Arnold-Chiari syndrome with flaccid paralysis and huge syringomyelia. AB - STUDY DESIGN: A case report. SETTING: Department of Orthopaedic Surgery, Shiga University of Medical Science, Japan. PATIENT: A 13-year-old woman presented progressive weakness in the lower extremities, with predominance on the right. Magnetic resonance (MR) imaging revealed a huge syrinx. The patient also showed scoliosis, cleft palate, hearing impairment, excessive sweating, hairiness, dural ectasia, and malformation of the skull. METHOD AND OBJECTIVES: We treated a very rare case of Arnold-Chiari syndrome, which presented with flaccid paralysis. Methods of differential diagnosis and suitable treatment are discussed. RESULTS AND CONCLUSION: Both the syrinx and muscle strength were quickly improved following placement of a syringo-peritoneal (S-P) shunt, after which the patient recovered the ability to walk. However, transient hypesthesia in the right hand occurred after the operation. The syrinx around the conus was thought to play a crucial role in the etiology of the patient case, which showed unique symptoms. PMID- 15111991 TI - Two case study reports of sleep apnoea in patients with paraplegia. AB - STUDY DESIGN: Case studies of sleep apnoea occurring in two patients with paraplegia. OBJECTIVE: To raise awareness of sleep apnoea in paraplegia. SETTING: Belfast, Northern Ireland. CASE REPORT: We report two patients with paraplegia, one who was having apparent episodes of loss of consciousness and the other daytime somnolence, who were found to have sleep apnoea. The first patient had been medically investigated extensively and a diagnosis of epilepsy was being considered. A joint consultation with the respective partners in each case revealed periods of night-time apnoea and led to sleep study investigations. CONCLUSION: Sleep apnoea is a treatable condition that can occur in patients with paraplegia who are not necessarily obese. Once diagnosed, resolution of symptoms can be rapid and can result in improved quality of life for patients. PMID- 15111992 TI - Use of the ICF conceptual framework to interpret hand function outcomes following tendon transfer surgery for tetraplegia. AB - STUDY DESIGN: Clinical commentary OBJECTIVE AND SETTING: This paper is a clinical commentary based on the Round Table discussion on Assessment and Outcomes at the 7th International Conference on Tetraplegia: Surgery and Rehabilitation, Bologna, Italy 6-8 June, 2001. It refers specifically to the 10-year re-review undertaken in 2001 at the Spinal Unit, Burwood Hospital, Christchurch, New Zealand. SUBJECTS: In all, 24 tetraplegic persons at a minimum of 12 years and up to 18 years following bilateral forearm tendon transfer surgery. METHOD: The data were interpreted using the International Classification of Functioning, Disability, and Health (ICF) conceptual framework as the basis of interdisciplinary understanding of the participation dimension. RESULTS: The results of the study outlined confirm that outcome measurement at more than one level of functioning is desirable to determine the functional effects beyond grip strength levels and activities of daily living, to consider the dimension of participation. CONCLUSIONS: Use of the ICF as a theoretical framework for interpretation of the results enhanced the clinical applicability of the outcome measures used in the 10-year re-review undertaken in New Zealand in 2001. PMID- 15111993 TI - A new test to improve the training quality of wheelchair racing athletes. AB - STUDY DESIGN: Validation of a new wheelchair racing test. OBJECTIVE: To assess the reproducibility of test parameters at different, subjectively chosen, intensity levels in wheelchair racing athletes. SETTING: Swiss Paraplegic Centre, Nottwil, Switzerland. SUBJECTS: We tested 11 (eight male, three female) young competitive wheelchair racing athletes. Their age was 20.5+/-6.4 years, height 164.7+/-16.3 cm and weight 54.1+/-9.2 kg. Average weekly training time was 6.8+/ 2.7 h. All had been engaged in regular training for over 3.9+/-2.8 years. METHODS: Within a period of 3+/-1 days, every athlete completed two identical tests in their own racing chair on a training roller. The tests consisted of five 1500 m bouts at routine training intensities of 1 (warm-up) up to 5 (race speed), with a rest of 2 min between each bout. The athletes were blinded to all the collected data during the whole of the test, except for indications of the 500, 1000 and 1500 m markers. We measured the overall time (for 1500 m), average speed, stroke frequency, heart rate, rate of perceived exertion (RPE) and the concentration of lactic acid for all five intensity bouts. In order to get a measure on how reproducible these intensity levels were, we compared the two tests of each athlete with each other, and calculated the root-mean-squared coefficients of variation (CV) for all measured parameters during every bout. RESULTS: CVs of the measured data show that the most reproducible values were found for bout 5 (2.6-7.9%); except for the lactic acid parameter. The heart rate (CV: 3.1-6.4%) and stroke frequency (CV: 6.5-7.9%) parameters reached rather constant values throughout all five bouts. Lactic acid concentrations showed very high CVs (16.8-29.7%). CONCLUSION: We conclude that this test is, with the exception of the lactic acid measurements, well reproducible, and particularly suitable for young wheelchair athletes. We find it to be a helpful tool for improving awareness for the individual training intensities, and for pursuing the development of the training process, as particularly the high-intensity bouts are well reproducible. PMID- 15111994 TI - Magnetic brain stimulation can improve clinical outcome in incomplete spinal cord injured patients. AB - STUDY DESIGN: Preliminary longitudinal clinical trial. OBJECTIVES: To test the efficacy of repetitive transcranial magnetic stimulation (rTMS) in modulating corticospinal inhibition and improving recovery in stable incomplete spinal cord injury (iSCI). SETTING: National Spinal Injuries Centre, Stoke Mandeville Hospital, Bucks, UK and Division of Neuroscience, Imperial College Faculty of Medicine, Charing Cross Hospital, London, UK. METHODS: Four stable iSCI patients were treated with rTMS over the occipital cortex (sham treatment) and then over the motor cortex (real treatment). Patients were assessed using electrophysiological, clinical and functional measures before treatment, during sham treatment, during the therapeutic treatment and during a 3-week follow-up period. RESULTS: Cortical inhibition was reduced during the treatment week. Perceptual threshold to electrical stimulation of the skin, ASIA clinical measures of motor and sensory function and time to complete a peg-board improved and remained improved into the follow-up period. CONCLUSION: In this preliminary trial, rTMS has been shown to alter cortical inhibition in iSCI and improve the clinical and functional outcome. SPONSORSHIP: This work was supported by the International Spinal Research Trust. PMID- 15111995 TI - Pyrexia due to pyogenic sacroiliitis with iliopsoas abscess after spinal cord injury. AB - STUDY DESIGN: Single case report. OBJECTIVES: To present an unusual cause of fever in a patient with spinal cord injury (SCI). SETTING: University Hospital, Belgium. METHODS: A 52-year-old man with a complete T9 paraplegia was admitted to hospital with a 7 day history of fever above 39 degrees C without pain and without gastrointestinal, urinary, or respiratory complaints. The patient had had a flap coverage for a sacral pressure ulcer 6 months prior to admission. RESULTS: Bone scintigraphy demonstrated markedly increased activity in the left sacroiliac joint. Computed tomography (CT) revealed an infection of the left sacroiliac joint with a large abscess involving the iliopsoas muscle. The responsible organism, Pseudomonas aeruginosa, was isolated from abscess liquid obtained by CT guided aspiration. We postulated that P. aeruginosa had colonized the eschar and, due to the proximity, infected the sacroiliac joint and the adjacent iliopsoas muscle. Prompt intravenous antibiotic therapy ensured clinical improvement and radiological regression. CONCLUSION: Pyogenic sacroiliitis is a relatively rare condition that may be difficult to diagnose in patients with normal sensation, and even more so in SCI patients. As far as we know, psoas abscess associated with pyogenic sacroiliitis has never been described in SCI patients. This infectious pathology must be kept in mind in SCI patients with fever of unknown origin and with a history of sacral eschar. PMID- 15111996 TI - Life situation of spinal cord-injured persons in Central Finland. AB - STUDY DESIGN: Individual interview and questionnaire of a regional spinal cord injured (SCI) population. OBJECTIVE: Local health care centres are mainly responsible for the SCI patients health care and rehabilitation after initial hospitalization in Finland. The purpose of the present study was to study aspects of the SCI patients life situation and their opinion of the health care services after 1 year since the injury. SETTING: Regional study in Central Finland. METHODS: Subjects were individually interviewed in their homes using a semi structured questionnaire, the Functional Independence Measure (FIM) and the Beck Depression Inventory (BDI). RESULTS: The female/male ratio was 1:4. The mean age at the time of injury was 34 years in female and 40 in male. Traffic accidents accounted for 46% of the injuries. In all, 48 subjects (63%) used electrical or manual wheelchair or both. Most subjects regarded their living conditions as good having enough personal assistance in their everyday life. Only 10% of the subjects were employed. Half of the subjects had had out-patient physiotherapy, but no other therapies after 1 year since the injury. More than half of the subjects were dissatisfied with the current health care services. Many had experienced complications after the initial hospitalization, especially urinary tract infections, decubitus ulcers and neurogenic-type pain. FIM classified well according to the disability groups. The motor complete tetraplegic group had the lowest mean scores (63) and the recovered group the highest ones (122). There was no association between the FIM and the BDI results. There was a correlation between the BDI and the subject's age at the time of the injury and the year of the injury. The older the subjects were when injured, the higher were the BDI scores, that is, they had more depressive symptoms. Those injured in the 1990s had the highest BDI scores. Nearly one-third of the subjects had mild, moderate or severe depression. CONCLUSIONS: The reported medical complications, depression and dissatisfaction with the health care services support a life-long care for SCI patients in Jyvaskyla Central hospital, not in the local health care centres. The psychological services, for example, prevention and treatment of depression, in particular, require more attention. PMID- 15111997 TI - Idiopathic spontaneous spinal subarachnoid hemorrhage. AB - INTRODUCTION: Spontaneous spinal subarachnoid hemorrhage (SAH) occurs in less than 1% of all cases of SAH, and idiopathic spontaneous spinal SAH is even more rare. METHODS: A 48-year-old man presented with a sudden onset of low back pain without any neurological deficit. There was no history of either trauma or the use of anticoagulants. Magnetic resonance (MR) imaging of the lumbar spine revealed an intradural linear mass from T12 to L3, ventral to the conus medullais and cauda equina. The mass was of iso-signal intensity in T2-weighted images, of high signal intensity in T1-weighted images, and was not enhanced following injection of gadolium. Lumbar tapping revealed bloody cerebrospinal fluid, confirming SAH. At 1 month after the onset of symptom, his pain subsided spontaneously and no vascular abnormality was found by spinal angiography. Follow up MR imaging confirmed the complete resolution of the SAH. CONCLUSION: We report a case of idiopathic spontaneous spinal SAH at the lumbar level in a 48-year-old man. PMID- 15111998 TI - Effectiveness of FK506 on lipid peroxidation in the spinal cord following experimental traumatic injury. AB - STUDY DESIGN: An in vivo study in Wistar albino rats with injured spinal cord. SETTING: Department of Neurosurgery, Biochemistry and Pathology, Gazi University, Ankara, Turkey. OBJECTIVES: The aim of this study was to investigate and compare the effects of FK506 an immunosupressive agent with methylprednisolone (MP) on lipid peroxidation (LP) in injured spinal cord tissue. METHOD: A total of 28 adult healthy Wistar albino rats were subjected to traumatic spinal cord injuries (SCI) by using an aneurysmal clip compression technique, and they were divided into four groups. The G1 group (n=8) received FK506 (1 mg/kg); the G2 group (n=8) received FK506 (1 mg/kg) and MP (30 mg/kg); the G3 group (n=6) received only MP (30 mg/kg); and the G4 group (n=6) received no medication. The injured spinal cord tissue was studied by means of lipid peroxides, malondialdehyde (MDA), with thiobarbituric acid reaction and additionally the FK506 (G1); the MP (G3) groups were studied for histopathologic alterations 72 h after SCI with eight separate animals. RESULTS: Although LP values of G1, G2, G3 showed no statistical difference between intergroup analyses (P=0.547), a histopathological examination revealed that in the group that received MP, the oedema pattern was more significant than the group that received FK506. Another interesting finding was the presence of polymorphonuclear leucocytes in the MP group, whereas no infiltration was found in the FK506 group. CONCLUSION: Analysis of the results indicated that FK506 is a valuable pharmacological agent that could be used to decrease the LP and polymorphonuclear leucocyte infiltration and inflamatory reactions in the injured spinal cord tissue. PMID- 15111999 TI - Pressure sores and blood and serum dysmetabolism in spinal cord injury patients. AB - STUDY DESIGN: Spinal cord injury (SCI) patients with pressure sores were studied before and after surgical intervention for ulcer healing and compared with matched SCI patients without sores and with patients with pressure sores and other diseases. OBJECTIVE: To analyse the relationship between pressure sores and anaemia and serum protein alteration in SCI patients. To study the pathogenesis of these alterations and suggest appropriate therapy. SETTING: Spinal cord unit in Rome, Italy. SUBJECTS: A total of 13 SCI patients with pressure sores, 13 comparable patients without pressure sores and four patients with other diseases and pressure sores. MAIN MEASURES: Haematochemical parameters. RESULTS: Patients with pressure sore showed significant decreased red cells, decreased haemoglobin and haematocrit, increased white cells and ferritin and decreased transferrin and transferrin saturation; total hypoproteinemia and hypoalbuminemia with increased Alfa-1 and gamma globulins increased erythrocyte sedimentation rate and C reactive protein were also present. The alterations returned to normal after surgical intervention for pressure sore healing. CONCLUSIONS: Patients with pressure sores suffer from anaemia and serum protein alteration that fells within the range of metabolic alteration of chronic disorders and neoplastic diseases. The alterations depend on a decreased utilisation of iron stores in the reticuloendothelial system and on inhibition of the hepatic synthesis of albumin. With regard to treatment, iron treatment should be avoided because of the risk of haemochromatosis. PMID- 15112000 TI - Unilateral cervical facet fracture: presentation of two cases and literature review. AB - STUDY DESIGN: Two patients with diagnosis of unilateral cervical facet fracture due to motor vehicle accident (MVA) are presented, and the literature is reviewed. OBJECTIVE: To discuss the diagnostic difficulties and management strategies in two patients with post-traumatic cervical facet fracture. SETTING: Department of Neurosurgery, Zonguldak Karaelmas University, Faculty of Medicine, Turkey. SUBJECT: Nonoperative treatment with immobilization was preferred in two female cases (33-34 years old) with diagnosis of C6-7 facet fracture following MVA. Magnetic resonance imaging (MRI) could be performed in acute period in the first case, but not in the second because of inadequate technical condition. RESULT: The first case with a good compliance to immobilization recovered without any neurological complication. However, the second case mobilized earlier and used a collar irregularly. Instability developed in the second case on the second month and surgical intervention with anterior approach was performed. CONCLUSION: The diagnosis of unilateral facet fractures is often missed and the treatment is still controversial. The compliance of the patient to cervical immobilization in nonoperative treatment plays a very important role in the development of late complications. MRI in the acute period may be useful in determining instability. PMID- 15112001 TI - Feeding the infants of HIV-infected mothers. PMID- 15112002 TI - Gender and the 10/90 gap in health research. PMID- 15112003 TI - The role of journals in enhancing health research in developing countries. PMID- 15112004 TI - Are WHO/UNAIDS/UNICEF-recommended replacement milks for infants of HIV-infected mothers appropriate in the South African context? AB - OBJECTIVE: Little is known about the nutritional adequacy and feasibility of breastmilk replacement options recommended by WHO/UNAIDS/UNICEF. The study aim was to explore suitability of the 2001 feeding recommendations for infants of HIV infected mothers for a rural region in KwaZulu Natal, South Africa specifically with respect to adequacy of micronutrients and essential fatty acids, cost, and preparation times of replacement milks. METHODS: Nutritional adequacy, cost, and preparation time of home-prepared replacement milks containing powdered full cream milk (PM) and fresh full cream milk (FM) and different micronutrient supplements (2 g UNICEF micronutrient sachet, government supplement routinely available in district public health clinics, and best available liquid paediatric supplement found in local pharmacies) were compared. Costs of locally available ingredients for replacement milk were used to calculate monthly costs for infants aged one, three, and six months. Total monthly costs of ingredients of commercial and home-prepared replacement milks were compared with each other and the average monthly income of domestic or shop workers. Time needed to prepare one feed of replacement milk was simulated. FINDINGS: When mixed with water, sugar, and each micronutrient supplement, PM and FM provided <50% of estimated required amounts for vitamins E and C, folic acid, iodine, and selenium and <75% for zinc and pantothenic acid. PM and FM made with UNICEF micronutrient sachets provided 30% adequate intake for niacin. FM prepared with any micronutrient supplement provided no more than 32% vitamin D. All PMs provided more than adequate amounts of vitamin D. Compared with the commercial formula, PM and FM provided 8-60% of vitamins A, E, and C, folic acid, manganese, zinc, and iodine. Preparations of PM and FM provided 11% minimum recommended linoleic acid and 67% minimum recommended alpha-linolenic acid per 450 ml mixture. It took 21-25 minutes to optimally prepare 120 ml of replacement feed from PM or commercial infant formula and 30-35 minutes for the fresh milk preparation. PM or FM cost approximately 20% of monthly income averaged over the first six months of life; commercial formula cost approximately 32%. CONCLUSION: No home-prepared replacement milks in South Africa meet all estimated micronutrient and essential fatty acid requirements of infants aged <6 months. Commercial infant formula is the only replacement milk that meets all nutritional needs. Revisions of WHO/UNAIDS/UNICEF HIV and infant feeding course replacement milk options are needed. If replacement milks are to provide total nutrition, preparations should include vegetable oils, such as soybean oil, as a source of linoleic and alpha-linolenic acids, and additional vitamins and minerals. PMID- 15112005 TI - Criteria for priority-setting in health care in Uganda: exploration of stakeholders' values. AB - OBJECTIVE: To explore stakeholders' acceptance of criteria for setting priorities for the health care system in Uganda. METHODS: A self-administered questionnaire was used. It was distributed to health workers, planners and administrators working in all levels of the Ugandan health care system. It was also distributed to members of the public. Participants were asked how strongly they agreed or disagreed with 18 criteria that could be used to set priorities for allocating health care. A total of 408 people took part. Data were entered and analysed using SPSS statistical software. Predetermined cut-off points were used to rank the criteria into three different categories: high weight (>66% of respondents agreed), average weight (33-66% of respondents agreed) and low weight (<33% of respondents agreed). We also tested for associations between respondents' characteristics and their degree of agreement with the criteria. FINDINGS: High weight criteria included severity of disease, benefit of the intervention, cost of the intervention, cost-effectiveness of the intervention, quality of the data on effectiveness, the patients age, place of residence, lifestyle, importance of providing equity of access to health care and the community's views. The average weight criteria included the patient's social status, mental features, physical capabilities, political views, responsibilities for others and gender. Low-weight criteria included the patient's religion, and power and influence. There were few associations between respondents' characteristics and their preferences. CONCLUSION: There was a high degree of acceptance for commonly used disease related and society-related criteria. There was less agreement about the patient related criteria. We propose that average-weight criteria should be debated in Uganda and other countries facing the challenge of distributing scarce health care resources. PMID- 15112006 TI - Condom use within marriage: a neglected HIV intervention. AB - OBJECTIVE: To assess the contraceptive effectiveness of condoms versus oral contraceptive pills and estimate the reproductive consequences of a major shift from pill to condom use. METHODS: Secondary analysis was performed on nationally representative cross-sectional surveys of women in 16 developing countries. FINDINGS: In the 16 countries, the median per cent of married couples currently using condoms was 2%, compared with 13% for the pill. Condom users reported a higher 12-month failure and higher method-related discontinuation rates than pill users (9% and 44% vs 6% and 30%, respectively). Condom users were more likely to report subsequent abortion following failure (21% vs 14%), and also more likely to switch rapidly to another method (76% vs 58%). The reproductive consequences, in terms of abortion and unwanted births, of a hypothetical reversal of the relative prevalence of condom and pill were estimated to be minor. The main reason for this unexpected result is that the majority of abortions and unwanted births arise from non-use of any contraceptive method. CONCLUSION: A massive shift from the more effective oral contraceptive pills to the less effective condom would not jeopardize policy goals of reducing abortions and unwanted births. However, such a shift would potentially have an added benefit of preventing human immunodeficiency virus (HIV) infections, especially in countries with generalized HIV epidemics. PMID- 15112007 TI - Discontinuation of cost sharing in Uganda. AB - OBJECTIVE: To assess the effects of ending cost sharing on use of outpatient services and how this was perceived by health workers and members of a health unit management committee. METHODS: From 10 districts across Uganda, 78 health facilities were selected. Attendance at these facilities was assessed for eight months before and 12 months after cost sharing ended. The data represented 1 966 522 outpatient visits. Perceptions about the impact of ending cost sharing were obtained from the 73 health workers and 78 members of the health unit management committee who were available. FINDINGS: With the end of cost sharing, the mean monthly number of new visits increased by 17 928 (53.3%), but among children aged <5 years the increase was 3611 (27.3%). Mean monthly reattendances increased by 2838 (81.3%) among children aged <5 years and 1889 (24.3%) among all people. Attendances for immunizations, antenatal clinics, and family planning all increased, despite these services having always been free. Health workers reported a decline in morale, and many health unit management committees no longer met regularly. CONCLUSION: Use of all services increased - even those that had never before been subject to fees. The loss of some autonomy by the health facility and diminished community governance of health facilities may have long term negative effects. PMID- 15112008 TI - Effectiveness of residual spraying of peridomestic ecotopes with deltamethrin and permethrin on Triatoma infestans in rural western Argentina: a district-wide randomized trial. AB - OBJECTIVE: To compare the effectiveness of a single residual spraying of pyrethroids on the occurrence and abundance of Triatoma infestans in peridomestic ecotopes in rural La Rioja. METHODS: A total of 667 (32.8%) peridomestic sites positive for T. infestans in May 1999 were randomly assigned to treatment within each village, sprayed in December 1999, and reinspected in December 2000. Treatments included 2.5% suspension concentrate (SC) deltamethrin in water at 25 mg active ingredient (a.i.)/m(2) applied with: (a) manual compression sprayers (standard treatment) or (b) power sprayers; (c) 1.5% emulsifiable concentrate (EC) deltamethrin at 25 mg a.i./m(2); and (d) 10% EC cis-permethrin at 170 mg a.i./m(2). EC pyrethroids were diluted in soybean oil and applied with power sprayers. All habitations were sprayed with the standard treatment. FINDINGS: The prevalence of T. infestans 1-year post-spraying was significantly lower in sites treated with SC deltamethrin applied with manual (24%) or power sprayers (31%) than in sites treated with EC deltamethrin (40%) or EC permethrin (53%). The relative odds of infestation and catch of T. infestans 1-year post-spraying significantly increased with the use of EC pyrethroids, the abundance of bugs per site before spraying, total surface, and host numbers. All insecticides had poor residual effects on wooden posts. CONCLUSION: Most of the infestations probably originated from triatomines that survived exposure to insecticides at each site. Despite the standard treatment proving to be the most effective, the current tactics and procedures fail to eliminate peridomestic populations of T. infestans in semiarid rural areas and need to be revised. PMID- 15112009 TI - The determinants of self-reported health-related quality of life in a culturally and socially diverse South African community. AB - OBJECTIVE: To determine factors predictive of the score on the visual analogue scale (VAS) of the EQ-5D questionnaire. METHODS: The responses of 1159 residents of a socially and ethnically diverse suburb of Cape Town, South Africa, to the EQ 5D questionnaire were analysed using forward stepwise multiple regression. The variables entered included ethnic group, religious affiliation (Christian or Muslim), income level, unemployment, recent illness or disability and each level of the five EQ-5D domains. FINDINGS: The model developed accounted for an adjusted r(2) of 0.234 and included 11 variables. In addition to the EQ-5D domains, the presence of a disability, an income of less than 420 US dollars per month, unemployment and age in years were significant predictors of VAS score. CONCLUSION: The substantial contribution of health state to the VAS indicates that it is a valid measure of health-related quality of life (HRQoL) across population groups. However, the subjects with lower social status reported a worse HRQoL than their health state alone warranted and this variable might need to be taken into account if the VAS is to be used to compare health states across populations. This paper provides empirical evidence of how HRQoL is perceived by different socioeconomic, cultural, ethnic and religious communities within a developing country. PMID- 15112010 TI - Tobacco smoking and oral clefts: a meta-analysis. AB - OBJECTIVE: To examine the association between maternal smoking and non-syndromic orofacial clefts in infants. METHODS: A meta-analysis of the association between maternal smoking during pregnancy was carried out using data from 24 case-control and cohort studies. FINDINGS: Consistent, moderate and statistically significant associations were found between maternal smoking and cleft lip, with or without cleft palate (relative risk 1.34, 95% confidence interval 1.25-1.44) and between maternal smoking and cleft palate (relative risk 1.22, 95% confidence interval 1.10-1.35). There was evidence of a modest dose-response effect for cleft lip with or without cleft palate. CONCLUSION: The evidence of an association between maternal tobacco smoking and orofacial clefts is strong enough to justify its use in anti-smoking campaigns. PMID- 15112011 TI - Population ageing in Lebanon: current status, future prospects and implications for policy. AB - During the past three decades, fast declines in fertility and mortality in Lebanon have created a compressed demographic transition, a growing trend towards survival into later life, and a larger proportion of elderly people in the population. Projections show that people aged 65 years and over are expected to constitute 10.2% of the population by 2025. Nevertheless, changes to the structure and composition of the population remain unmatched by any corresponding increase in support measures either through formal channels such as pension plans or through health or socioeconomic security measures such as the provision of subsidies for health care, home help or any form of nursing care. This means that an older person is forced to be dependent upon family support if it exists. We examine demographic trends of population ageing in Lebanon between 1970 and 1995 and provide projections until 2025. Variations in population ageing within the country are also considered. We also assess health care and social policy implications of demographic changes in the context of health and economic sector reforms initiated recently by the state, and explore their impact upon the expanding population of elderly people. PMID- 15112012 TI - Galvanizing mental health research in low- and middle-income countries: role of scientific journals. PMID- 15112013 TI - Vitamin and mineral deficiencies harm one-third of the world's population, says new report. PMID- 15112014 TI - US and Thai Governments defend HIV/AIDS vaccine trial in Thailand. PMID- 15112015 TI - Ailing French health care system to go under the knife. PMID- 15112016 TI - Top broadcasters join forces with UN on HIV/AIDS prevention. PMID- 15112020 TI - Flagging global sanitation target threatens other Millennium Development Goals. PMID- 15112021 TI - Avian flu virus could evolve into dangerous human pathogen, experts fear. PMID- 15112022 TI - WHO refutes malaria malpractice allegations. PMID- 15112023 TI - West Africa prepares for the final battle against polio. PMID- 15112024 TI - WHO issues guidelines for herbal medicines. PMID- 15112030 TI - Effects of nicotine and mecamylamine on cognition in rhesus monkeys. AB - RATIONALE: Nicotine and other agonists of nicotinic cholinergic receptors (nAChR) have been shown to improve performance in specific memory domains in rodents and monkeys. Such beneficial effects are observed in preclinical models of age related cognitive decline, stimulating interest in nAChR ligands as possible therapeutics. Prior work has typically focused on assays of spatial working memory in rodent studies and visual recognition memory in monkey studies. OBJECTIVE: The current study was conducted to determine the role played by nAChRs in multiple types of memory in monkeys. METHODS: Rhesus monkeys (n=6) were trained to perform a battery of six behavioral tasks and then serially challenged with acute doses of nicotine (3.2-56 microg/kg, i.m.) and the nAChR antagonist mecamylamine (0.32-1.78 mg/kg, i.m.). RESULTS: Nicotine improved performance on tests designed to assay visual recognition memory, spatial working memory and visuo-spatial associative memory, while mecamylamine impaired visuo-spatial associative memory. Ballistic and fine motor performance was not significantly improved by nicotine but fine motor performance was impaired by mecamylamine. CONCLUSIONS: Although nicotine may improve performance in multiple domains, effects on visuo-spatial associative memory is the most specifically attributable to nAChR signaling. PMID- 15112031 TI - Effects of the kappa opioid agonist U50,488 and the kappa opioid antagonist nor binaltorphimine on choice between cocaine and food in rhesus monkeys. AB - RATIONALE: Selective kappa opioid receptor agonists usually decrease cocaine self administration in procedures that use rate-based measures of reinforcement; however, the rate-altering effects of kappa agonists complicate interpretation of these findings. OBJECTIVES: To evaluate the effects of the selective kappa agonist U50,488 and the selective kappa antagonist nor-binaltorphimine (nor-BNI) on concurrent choice between cocaine and food in rhesus monkeys. The concurrent choice procedure provides a rate-independent measure of the relative reinforcing effects of cocaine in comparison with food. METHODS: Four rhesus monkeys were trained to respond under a concurrent-choice schedule for food (1-g pellets) or cocaine (0-0.1 mg/kg per injection). Saline and increasing doses of U50,488 (0.0032-0.1 mg/kg per h) were administered by pseudo-continuous i.v. infusion (one infusion every 20 min) during sequential 3-day blocks. In a separate experiment, monkeys were treated with nor-BNI (3.2 mg/kg, i.v.), and cocaine choice was re-determined during pseudo-continuous infusion with saline or U50,488 (0.1 mg/kg per h). RESULTS: During saline treatment, cocaine maintained a dose dependent and monotonic increase in cocaine choice. Monkeys responded primarily for food when low cocaine doses were available (0-0.01 mg/kg per injection) and primarily for cocaine when higher cocaine doses were available (0.032-0.1 mg/kg per injection). U50,488 produced a dose-dependent increase in cocaine choice, manifested as leftward shifts in the cocaine-choice, dose-effect curve. U50,488 also dose-dependently decreased overall response rates. Nor-BNI did not alter cocaine choice, but it attenuated the effects of U50,488. CONCLUSIONS: These results suggest that continuous treatment with U50,488 produces a kappa receptor mediated increase in the relative reinforcing effects of cocaine in comparison with food. PMID- 15112032 TI - Rapamycin-associated post-transplantation glomerulonephritis and its remission after reintroduction of calcineurin-inhibitor therapy. AB - Rapamycin is a new immunosuppressive agent approved for maintenance therapy after kidney transplantation. It may allow calcineurin-inhibitor-free, non-nephrotoxic immunosuppression. We report, however, on four kidney-transplant recipients who developed post-transplantation glomerulonephritis after conversion from a calcineurin-inhibitor-based immunosuppression to rapamycin. In all four patients nephrotic-range proteinuria occurred 2-9 months after conversion to rapamycin. Renal biopsy confirmed membrano-proliferative glomerulonephritis type 1 in one case, membranous glomerulonephritis in another and IgA-nephropathy in two cases, respectively. Calcineurin-inhibitor-based immunosuppression was reintroduced and resulted in complete remission of proteinuria and in stabilised renal function in all patients. We conclude that in the case of rapamycin-associated post transplantation glomerulonephritis an attempt should be made to replace rapamycin by a calcineurin inhibitor. PMID- 15112033 TI - Introducing Critical Care Outreach: a ward-randomised trial of phased introduction in a general hospital. AB - OBJECTIVE: The purpose of the study was to investigate the effects of introducing a critical care outreach service on in-hospital mortality and length of stay in a general acute hospital. DESIGN: A pragmatic ward-randomised trial design was used, with intervention introduced to all wards in sequence. No blinding was possible. SETTING: Sixteen adult wards in an 800-bed general hospital in the north of England. PATIENTS AND PARTICIPANTS: All admissions to the 16 surgical, medical and elderly care wards during 32-week study period were included (7450 patients in total, of whom 2903 were eligible for the primary comparison). INTERVENTIONS: Essential elements of the Critical Care Outreach service introduced during the study were a nurse-led team of nurses and doctors experienced in critical care, a 24-h service, emphasis on education, support and practical help for ward staff. MEASUREMENTS AND RESULTS: The main outcome measures were in-hospital mortality and length of stay. Outreach intervention reduced in-hospital mortality compared with control (two-level odds ratio: 0.52 (95% CI 0.32-0.85). A possible increased length of stay associated with outreach was not fully supported by confirmatory and sensitivity analyses. CONCLUSIONS: The study suggests outreach reduces mortality in general hospital wards. It may also increase length of stay, but our findings on this are equivocal. PMID- 15112035 TI - Psychiatric outcome following paediatric intensive care unit (PICU) admission: a cohort study. AB - OBJECTIVE: To determine whether paediatric intensive care unit (PICU) admission is associated with greater psychiatric morbidity in children and parents as compared with general paediatric ward admissions. DESIGN: Retrospective cohort study. SETTING: Paediatric intensive care unit and two general paediatric wards of a London teaching hospital. PARTICIPANTS: Children aged 5-18 years discharged from PICU (exposed cohort) and general paediatric wards (unexposed cohort) 6-12 months previously, together with their parents. MEASUREMENTS AND RESULTS: Children: the Clinician Administered Post Traumatic Stress Disorder (PTSD) Scale for Children (CAPS-C), the Impact of Event Scale (IES), Strengths and Difficulties Questionnaire, Birleson Depression Scale, Revised Children's Manifest Anxiety Scale, Child Somatization Inventory. Parents: IES, General Health Questionnaire, Beck Depression Inventory, Hospital Anxiety and Depression Scale. Thirty-five of 46 (76%) PICU-discharged families and 33 of 41 (80%) from general paediatric wards participated. Valid CAPS-C data were obtained for 19 PICU-admitted children and 27 children admitted only to the general paediatric ward; 4/19 (21%) of PICU-discharged children developed PTSD (compared with none of 27 ward admissions), p=0.02. PICU children had significantly more PTSD features of irritability and persistent avoidance of reminders of the admission. Parents of PICU children were more likely to screen positive for PTSD (9/33 (27%) compared with 2/29 (7%) parents of ward-admitted children), p=0.04. There were no significant differences between the groups for other measures of psychopathology. CONCLUSION: Post traumatic stress disorder diagnosis and symptomatology is significantly more common in families where a child has been admitted to the PICU. Consideration should be given to providing psychological support for children and parents after PICU admission. PMID- 15112034 TI - Antibiotic-mediated release of tumour necrosis factor alpha and norharman in patients with hospital-acquired pneumonia and septic encephalopathy. AB - OBJECTIVE: To investigate antibiotic-mediated release of tumour necrosis factor (TNF)-alpha and norharman in patients with hospital-acquired pneumonia with and without additional septic encephalopathy. DESIGN: Prospective observational study with a retrospective post hoc analysis. SETTING: Surgical intensive care unit (ICU) at a university hospital. PATIENTS: Thirty-seven patients were consecutively included (9 patients with hospital-acquired pneumonia, 11 patients with hospital-acquired pneumonia and septic encephalopathy, 17 control patients) in the study. Pneumonia was defined according to the criteria of the American Thoracic Society. INTERVENTIONS: Patients received cephalosporins for antibiotic treatment of hospital-acquired pneumonia. Blood samples were taken before, immediately after and 4 h after application of cephalosporins. MEASUREMENTS AND RESULTS: Of the pneumonia patients, 55% developed septic encephalopathy. ICU stay, complications and mortality were significantly increased. An increased release of TNF-alpha was immediately seen in all pneumonia patients after antibiotics compared to controls, whereas the level did not differ between patients with and without septic encephalopathy. Norharman was significantly increased in pneumonia patients 4 h after antibiotic treatment, in tendency more enhanced in the pneumonia patients without encephalopathy. CONCLUSIONS: Patients with hospital-acquired pneumonia and septic encephalopathy had a significantly longer ICU stay with higher mortality rate compared to patients with hospital acquired pneumonia alone. Antibiotic-mediated TNF-alpha release may induce the kynurenine pathway. TNF-alpha activates indolamine-2,3-dioxygenase with neurotoxic quinolinic acid as the end product. Norharman seems to counteract this mechanism and seems to play a role in neuroprotection. The worse outcome of patients with encephalopathy expresses the need to investigate protective factors and mechanisms. PMID- 15112036 TI - [The arthroscopic "wafer procedure" in degenerative disc ulnocarpal tears with ulnocarpal compression syndrome. Techniques, indications, results]. AB - The wafer procedure is a technique involving the partial resection of the distal ulna for the treatment of patients with symptomatic tears of the TFCC, for ulnar abutment syndrome or both. The TFCC-tears are classified as Palmer type 2. The wafer procedure can be performed as an open procedure or arthroscopically. It is an alternative to a shortening osteotomy of the ulna and decompresses the ulnocarpal joint. In ten cases with long-term follow-up, the preferability of the arthroscopic method is demonstrated: a minimally invasive technique, optimal assessment of all lesions, maximum protection of all uninjured structures in comparison to the open method, single stage procedure, and low complication rate. The long-term results are predominantly positive, so that the arthroscopic wafer procedure should be performed more often than it is today. PMID- 15112037 TI - Marker-assisted introgression of five QTLs controlling fruit quality traits into three tomato lines revealed interactions between QTLs and genetic backgrounds. AB - The evaluation of organoleptic quality of tomato fruit requires physical, chemical and sensory analyses, which are expensive and difficult to assess. Therefore, their practical use in phenotypic selection is difficult. In a previous study, the genetic control of several traits related to organoleptic quality of fresh-market tomato fruit was investigated. Five chromosome regions strongly involved in organoleptic quality attributes were then chosen to be introgressed into three different recipient lines through marker-assisted selection. A marker-assisted backcross (MABC) strategy was performed, as all the favorable alleles for quality traits were provided by the same parental tomato line, whose fruit weight (FW) and firmness were much lower than those of the lines commonly used to develop fresh market varieties. Three improved lines were obtained after three backcrossing and two selfing generations. The implementation of the MABC scheme is described. The three improved lines were crossed together and with the recipient lines in a half-diallel mating scheme, and the simultaneous effect of the five quantitative trait locus (QTL) regions was compared in different genetic backgrounds. Significant effects of the introgressed regions and of the genetic backgrounds were shown. Additive effects were detected for soluble solid and reducing sugar content in two genetic backgrounds. A partially dominant effect on titratable acidity was detected in only one genetic background. In contrast, additive to dominant unfavorable effects of the donor alleles were detected for FW and locule number in the three genetic backgrounds. Recessive QTL effects on firmness were only detected in the two firmest genetic backgrounds. Comparison of the hybrids in the half-diallel gave complementary information on the effects of: (1) the alleles at the selected regions, (2) the genetic backgrounds and (3) their interaction. Breeding efficiency strongly varied according to the recipient parent, and significant interactions between QTLs and genetic backgrounds were shown for all of the traits studied. PMID- 15112038 TI - [Outpatient extracorporeal shock wave lithotripsy. Prospective evaluation of 2937 cases]. AB - PURPOSE: Extracorporeal shock wave lithotripsy (ESWL) is mainly performed on inpatient basis in Germany. It has yet to be considered if outpatient treatment in Germany is indicated when sufficient urological care is guaranteed. MATERIALS AND METHODS: Between September 1989 and December 2002, a total of 10,505 patients with urolithiasis were treated at the Institut zur Steinzertrummerung in Troisdorf, Germany with the Siemens Lithostar plus. Of these patients, 2937 (43%) were treated on an outpatient basis. The data were collected prospectively from September 1989. Patient satisfaction was evaluated with a self-developed, non validated questionnaire. RESULTS: The overall success rate was 81.8%. The complication rate after ESWL was 40.2%, whereas 4.9% of the patients had to be admitted to a hospital; 73.5% did not need auxiliary treatment before and 78.6% after ESWL, respectively. More than 90% of the patients were satisfied with the result of outpatient treatment. CONCLUSIONS: Outpatient ESWL can be safely performed as minimally invasive treatment after thorough patient selection in Germany. The success and quality of treatment was not altered because of outpatient treatment. A network of ESWL center and post-interventional care center has to be established to guarantee 24-h service for auxiliary necessities. PMID- 15112039 TI - [Treatment options for bladder disorders in the aged]. AB - Urinary incontinence in the elderly is caused primarily by multiple factors such as physiologic changes of old age connected with multimorbidity and functional deficiencies, polypharmacology, psychosociological influences as well as conditions of the milieu. All these factors have to be carefully considered for therapy to be effective. Continence should be the goal of such therapy. Active participation in daily life can also be achieved by using auxiliary devices to improve the impaired quality of life. Temporary incontinence demands the search for its cause, while chronic incontinence calls for a differentiated form of therapy according to the kind of incontinence encountered. Conservative treatment consists of behavior therapy, such as toilet-(habit)-training and pelvic floor exercises. The use of drugs and auxiliary devices are equally important. PMID- 15112040 TI - [Changes in the receptor profile of the aging bladder]. AB - Future demographic developments will challenge urology with a steadily increasing incidence of lower urinary tract symptoms (LUTS) derived from the aging bladder. Obstruction, instability and hypocontractility, which may be caused by changes in the receptor profile of the detrusor, are typical pathophysiologic findings in geriatric bladder dysfunction. Benign prostatic hyperplasia and diabetes mellitus are age-associated comorbidities with an additional influence on bladder receptors. Muscarinic (M(2), M(3)), purinergic (P2X, P2Y) and adrenergic receptors (alpha(1), beta(3)) are targets of efferent sympathetic and parasympathetic bladder innervation. Although the results from animal experiments are somewhat inconsistent, aging and bladder outlet obstruction (BOO) probably cause partial cholinergic denervation of the detrusor with a subsequent upregulation of muscarinic receptor sensitivity leading to bladder instability. The non-cholinergic (atropine-resistant) component of the detrusor contraction rises with aging and BOO to 50%, emphasizing the increasing impact of purinergic receptors in geriatric LUTS. alpha(1)-adrenergic receptors are modulated in the aging bladder by a shift from the predominant alpha(1a) subtype to the alpha(1d) subtype, which shows 100-fold higher affinity towards norepinephrine and increases alpha-adrenergic bladder susceptibility. No data are available on the changes in beta(3) receptor density or sensitivity with aging. Moreover, the role of sensory C-fiber receptors in geriatric LUTS remains completely unclear, although specific C-fiber blockers are already under clinical evaluation (capsaicin, resiniferatoxin). PMID- 15112041 TI - [Necessity and usefulness of bioinformatic methods for microarray data analysis]. AB - Data emerging from DNA microarray experiments are usually difficult to interpret. While the level of expression of several thousand genes can be measured in a single experiment, only a few dozen experiments are normally carried out, leading to data sets of very high dimensionality and low cardinality. The computational analysis of gene expression data makes significant usage of machine learning and statistical methods. Nevertheless, caution should be used in the blind adoption of these methods, as this usually leads to an over-interpretation of the expression profiles. The following presentation provides an overview of up-to date principles of biostatistical analysis. A potential application for the analysis of high-dimensional expression profiles of prostate cancer is given. PMID- 15112042 TI - [Diagnostic imaging of pulmonary lymphangiosis carcinomatosis]. AB - The diagnosis of pulmonary lymphangiosis carcinomatosa (PLC) is of great importance for the prognostically-oriented therapy stratification of tumor patients. In this field, high-resolution computed tomography (HRCT) is the state of the art in imaging. Using HRCT, it is possible to identify pulmonary parenchymal structures in a detailed fashion to evaluate interstitial patterns. This step is preceded by an x-ray of the thorax that detects pathological findings and rules out other diseases. The typical characteristics of PLC are described with particular attention to HR-phenomenology, and discussed in comparison with the literature regarding anatomy and pathogenesis. Finally, conclusions are drawn for differential diagnosis and supported by characteristic cases. PMID- 15112043 TI - [Pathology, classification, and staging of malignant lung tumors]. AB - The histopathogical, immunohistochemical, and genetic characterization of specimens of, mostly advanced, lung tumors that show variable phenotypes in biopsies of just 1-2 mm does not allow conclusions regarding causal factors (e.g., smoking, radon, asbestos etc.) or further progress of the disease. Therapeutical approach and the still unfavorable prognosis remain essentially, as in the last thirty years, to be characterized by TNM and performance status of the individual patient and, to a lesser extent, by the main histological type of tumor. In recent years, our knowledge of the quite variable biology of tumors has been significantly increased by the use of immunohistochemical methods and molecular biology. These methods facilitated an improved qualitative and quantitative characterization of heterogeneously differentiated lung tumors (e.g., neuroendocrine/blastomatoid portions etc.). The detection of genetic alterations of tumor suppressor genes and oncogenes is, at the moment, only of scientific interest. The heterogeneity of tumors is emphasized by results obtained by molecular genetic techniques. A connection between the detected genetic anomalies and histomorphological growth patterns can not be seen. At the present time, the validity of individual findings for a correlation between operability, tumor progress, chemotherapy and prognosis is not sufficiently elucidated by investigations nor secured. PMID- 15112045 TI - [Optimization of staging in colon cancer using sentinel lymph node biopsy]. AB - Routine determination of the nodal status in colon cancer is strongly dependent on the individual quality and technique of histopathological assessment and surgical lymph node dissection. We evaluated whether sentinel lymph node biopsy (SLNB) could contribute to an improvement in staging. At least one SLN (median n=2) was detected (detection rate 84%) in each of 38 of 45 patients with primary colon cancer. Ten of these 38 were found to have lymph node metastases by HE staining (26%), six of them in the SLN. Nine of the 28 patients that were initially nodal-negative by HE revealed one micrometastasis and eight cases of isolated tumor cells by immunohistochemical (IHC) staining (32% upstaging response). Including the IHC-positive cases, 19 of the 38 patients were nodal positive (50%), 15 of them with tumor-infiltrated SLN (overall sensitivity of SLNB 79%). Using the dye method, SLNB is clinically practicable and leads in the majority of the patients to the detection of SLN. The selective, intensified histopathological assessment of SLN identifies small tumor cell deposits in a relevant percentage of patients with little and clinically practicable effort. PMID- 15112046 TI - [Five hundred outpatient hernioplasties using the Lichtenstein method]. AB - The Lichtenstein repair method is an excellent and simple technique for hernia repair performed as a day-case procedure. A consecutive series of 500 ambulatory groin hernia repairs from 1994 to 2002 was studied. The mean follow-up for 84.6% of the patients was 2 years (range 6-86 months). There were no severe complications. The rate of clinically important wound haematomas (n=6, 1.2%) was low, as well as the numbers of testicular atrophies (n=1, 0.2%) and deep wound infection (n=1, 0.2%). We saw no thrombosis. There were 15 recurrences (3.5%). Ninety-six per cent of the patients were satisfied with the outpatient operation. In day-case surgery, conditions for the patients' treatment at home should be checked carefully by surgeons before the operation. CONCLUSION: Our results suggest that the number of day-case hernia repairs can be increased without severe complications. The procedure can be performed on an outpatient basis without problems under general anaesthesia. Increasing day-case surgery could significantly reduce the costs of health care. PMID- 15112047 TI - [Paradigms in treatment of myocardial infarction]. PMID- 15112049 TI - Functional investigations of exercising muscle: a noninvasive magnetic resonance spectroscopy-magnetic resonance imaging approach. AB - Muscle fatigue, which is defined as the decline in muscle performance during exercise, may occur at different sites along the pathway from the central nervous system through to the intramuscular contractile machinery. Historically, both impairment of neuromuscular transmission and peripheral alterations within the muscle have been proposed as causative factors of fatigue development. However, according to more recent studies, muscle energetics play a key role in this process. Intramyoplasmic accumulation of inorganic phosphate (P(i)) and limitation in ATP availability have been frequently evoked as the main mechanisms leading to fatigue. Although attractive, these hypotheses have been elaborated on the basis of experimental results obtained in vitro, and their physiological relevance has never been clearly demonstrated in vivo. In that context, noninvasive methods such as 31-phosphorus magnetic resonance spectroscopy and surface electromyography have been employed to understand both metabolic and electrical aspects of muscle fatigue under physiological conditions. Mapping of muscles activated during exercise is another interesting issue which can be addressed using magnetic resonance imaging (MRI). Exercise-induced T2 changes have been used in order to locate activated muscles and also as a quantitative index of exercise intensity. The main results related to both issues, i.e. the metabolic and electrical aspects of fatigue and the MRI functional investigation of exercising muscle, are discussed in the present review. PMID- 15112050 TI - Novel aspects of glypican glycobiology. AB - Mutations in glypican genes cause dysmorphic and overgrowth syndromes in men and mice, abnormal development in flies and worms, and defective gastrulation in zebrafish and ascidians. All glypican core proteins share a characteristic pattern of 14 conserved cysteine residues. Upstream from the C-terminal membrane anchorage are 3-4 heparan sulfate attachment sites. Cysteines in glypican-1 can become nitrosylated by nitric oxide in a copper-dependent reaction. When glypican 1 is exposed to ascorbate, nitric oxide is released and participates in deaminative cleavage of heparan sulfate at sites where the glucosamines have a free amino group. This process takes place while glypican-1 recycles via a nonclassical, caveolin-1-associated route. Glypicans are involved in growth factor signalling and transport, e.g. of polyamines. Cargo can be unloaded from heparan sulfate by nitric oxide-dependent degradation. How glypican and its degradation products and the cargo exit from the recycling route is an enigma. PMID- 15112051 TI - Role of N-linked polymannose oligosaccharides in targeting glycoproteins for endoplasmic reticulum-associated degradation. AB - Misfolded or incompletely assembled multisubunit glycoproteins undergo endoplasmic reticulum-associated degradation (ERAD) regulated in large measure by their N-linked polymannose oligosaccharides. In this quality control system lectin interaction with Glc(3)Man(9)GlcNAc(2) glycans after trimming with endoplasmic reticulum (ER) alpha-glucosidases and alpha-mannosidases sorts out persistently unfolded glycoproteins for N-deglycosylation and proteolytic degradation. Monoglucosylated (Glc(1)Man(9)GlcNAc(2)) glycoproteins take part in the calnexin/calreticulin glucosylation-deglucosylation cycle, while the Man(8)GlcNAc(2) isomer B product of ER mannosidase I interacts with EDEM. Proteasomal degradation requires retrotranslocation into the cytosol through a Sec61 channel and deglycosylation by peptide: N-glycosidase (PNGase); in alternate models both PNGase and proteasomes may be either free in the cytosol or ER membrane-imbedded/attached. Numerous proteins appear to undergo nonproteasomal degradation in which deglycosylation and proteolysis take place in the ER lumen. The released free oligosaccharides (OS) are transported to the cytosol as OS GlcNAc(2) along with similar components produced by the hydrolytic action of the oligosaccharyltransferase, where they together with OS from the proteasomal pathway are trimmed to Man(5)GlcNAc(1) by the action of cytosolic endo-beta- N acetylglucosaminidase and alpha-mannosidase before entering the lysosomes. Some misfolded glycoproteins can recycle between the ER, intermediate and Golgi compartments, where they are further processed before ERAD. Moreover, properly folded glycoproteins with mannose-trimmed glycans can be deglucosylated in the Golgi by endomannosidase, thereby releasing calreticulin and permitting formation of complex OS. A number of regulatory controls have been described, including the glucosidase-glucosyltransferase shuttle, which controls the level of Glc(3)Man(9)GlcNAc(2)-P-P-Dol, and the unfolded protein response, which enhances synthesis of components of the quality control system. PMID- 15112052 TI - New approaches to therapy of cancers of the stomach, colon and pancreas based on peptide analogs. AB - Cancers of the stomach, colon and exocrine pancreas are major international health problems and result in more than a million deaths worldwide each year. The therapies for these malignancies must be improved. The effects of gastrointestinal (GI) hormonal peptides and endogenous growth factors on these cancers were reviewed. Some GI peptides, including gastrin and gastrin-releasing peptide (GRP) (mammalian bombesin), appear to be involved in the growth of neoplasms of the GI tract. Certain growth factors such as insulin-like growth factor (IGF)-I, IGF-II and epidermal growth factor and their receptors that regulate cell proliferation are also implicated in the development and progression of GI cancers. Experimental investigations on gastric, colorectal and pancreatic cancers with analogs of somatostatin, antagonists of bombesin/GRP, antagonists of growth hormone-releasing hormone as well as cytotoxic peptides that can be targeted to peptide receptors on tumors were summarized. Clinical trials on peptide analogs in patients with gastric, colorectal and pancreatic cancers were reviewed and analyzed. It may be possible to develop new approaches to hormonal therapy of GI malignancies based on various peptide analogs. PMID- 15112053 TI - The dual role of endothelial differentiation-related factor-1 in the cytosol and nucleus: modulation by protein kinase A. AB - Endothelial differentiation-related factor (EDF)-1 is involved in the repression of endothelial cell differentiation and is the first studied calmodulin (CaM) binding protein in endothelial cells. Here we report that (i) EDF-1 is in vitro and in vivo phosphorylated by protein kinase A (PKA); (ii) EDF-1/CaM interaction is modulated by the phosphorylation of EDF-1 by PKA; (iii) forskolin stimulates nuclear accumulation of EDF-1, and (iv) PKA phosphorylation enhances EDF-1 interaction with the TATA-binding protein. CaM modulates the activity of several enzymes, among which is nitric oxide synthase (NOS). EDF-1, but not phosphorylated EDF-1, inhibits the activity of NOS. Accordingly, we detected an increase in NOS activity in cells that express low amounts of EDF-1. Our results indicate that EDF-1 serves two main functions in endothelial cells: (i) it regulates CaM availability in the cytosol, and (ii) it acts in the nucleus as a transcriptional coactivator. PMID- 15112054 TI - Ubiquitin-proteasome pathway as a primary defender against TRAIL-mediated cell death. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces apoptotic cell death as well as expression of proinflammatory genes such as CXCL8 in malignant human astrocytoma cells. However, the molecular mechanisms that determine the fate of cells are not yet understood. The ubiquitin (Ub)-proteasome pathway regulates a wide range of cellular functions through degradation of various regulatory proteins; given this, we hypothesized that this pathway may play a central role in TRAIL-mediated signaling. We demonstrate here that inhibition of the Ub-proteasome pathway enhanced TRAIL-mediated cell death of human astrocytoma CRT-MG cells within hours by blocking degradation of active caspase-8 and -3. Proteasome inhibitors suppressed TRAIL-mediated activation of NF-kappaB; however, inhibition of the NF-kappaB pathway alone was not sufficient to enhance TRAIL-mediated cell death. Collectively, these results suggest that the Ub-proteasome pathway may play an important role as an antiapoptotic surveillance system by eliminating activated caspases as well as mediating NF kappaB-dependent signals. PMID- 15112055 TI - Myelination in the hippocampus during development and following lesion. AB - Myelin is crucial for the stabilization of axonal projections in the developing and adult mammalian brain. However, myelin components also act as a non permissive and repellent substrate for outgrowing axons. Therefore, one major factor which accounts for the lack of axonal regeneration in the mature brain is myelin. Here we report on the appearance of mature, fully myelinated axons during hippocampal development and following entorhinal lesion with the myelin-specific marker Black Gold. Although entorhinal axons enter the hippocampal formation at embryonic day 17, light and ultrastructural analysis revealed that mature myelinated fibers in the hippocampus occur in the second postnatal week. During postnatal development, increasing numbers of myelinated fibers appear and the distribution of myelinated fibers at postnatal day 25 was similar to that found in the adult. After entorhinal cortex lesion, a specific anterograde denervation in the hippocampus takes place, accompanied by a long-lasting loss of myelin. Quantitative analysis of myelin and myelin breakdown products at different time points after lesion revealed a temporally close correlation to the degeneration and reorganization pha-ses in the hippocampus. In contrast, electroconvulsive seizures resulted in brief demyelination and a faster recovery time course. In conclusion, we could show that the appearance of mature axons in the hippocampus is temporally regulated during development. In the adult hippocampus, demyelination was found after anterograde degeneration and also following seizures, suggesting that independent types of insult lead to demyelination. Reappearing mature axons were found in the hippocampus following axonal sprouting. Therefore, our quantitative analysis of mature axons and myelination effectively reflects the readjusted axonal density and possible electrophysiological balance following lesion. PMID- 15112056 TI - Opposite roles of protein kinase C isoforms in proliferation, differentiation, apoptosis, and tumorigenicity of human HaCaT keratinocytes. AB - We have previously shown that the protein kinase C (PKC) system plays a pivotal role in regulation of proliferation and differentiation of the human keratinocyte line HaCaT which is often used to assess processes of immortalization, transformation, and tumorigenesis in human skin. In this paper, using pharmacological and molecular biology approaches, we investigated the isoform specific roles of certain PKC isoenzymes (conventional cPKCalpha and beta; novel nPKCdelta and epsilon) in the regulation of various keratinocyte functions. cPKCalpha and nPKCdelta stimulated cellular differentiation and increased susceptibility of cells to actions of inducers of apoptosis, and they markedly inhibited cellular proliferation and tumor growth in immunodeficient mice. In marked contrast, cPKCbeta and nPKCepsilon increased both in vitro and in vivo growth of cells and inhibited differentiation and apoptosis. Our data present clear evidence for the specific, antagonistic roles of certain cPKC and nPKC isoforms in regulating the above processes in human HaCaT keratinocytes. PMID- 15112057 TI - Identification of swelling-activated Cl(-) current in rabbit cardiac Purkinje cells. AB - The presence and functional role of the swelling-activated Cl(-) current (I(Cl(swell))) in rabbit cardiac Purkinje cells was examined using patch-clamp methodology. Extracellular hypotonicity (210 or 135 mOsm) activated an outwardly rectifying, time-independent current with a reversal potential close to the calculated Cl(-) equilibrium potential (E(Cl)). The magnitude of this current was related to tonicity of the superfusate. The current was blocked by 0.5 mM 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). These features are comparable to those of I(Cl(swell)) found in sinoatrial nodal, atrial, and ventricular myocytes. I(Cl(swell)) activation at 210 and 135 mOsm depolarized the resting membrane potential with 6 and 10 mV and shortened the action potential by approximately 18 and approximately 33%, respectively. DIDS partially reversed I(Cl(swell))-induced action potential changes. We conclude that I(Cl(swell)) is present in Purkinje cells and its activation leads to action potential shortening and resting membrane potential depolarization, both of which can promote the development of reentrant arrhythmias. PMID- 15112058 TI - Progesterone receptors A and B differentially modulate corticotropin-releasing hormone gene expression through a cAMP regulatory element. AB - Corticotrophin-releasing hormone (CRH) plays a major role in mechanisms controlling human pregnancy and parturition. Gene regulation by progesterone may be a key point in the control of placental CRH production. Studies in primary placental cells show that antagonism of progesterone activity or production by RU486 or trilostane leads to an increase in CRH promoter activity. This effect can be reversed by the addition of progesterone. Overexpression of progesterone receptor A (PR-A) or glucocorticoid receptor resulted in a decrease in CRH promoter activity following progesterone treatment, whereas an increase in promoter activity was observed with overexpressed PR-B. Studies including mutation of the cAMP regulatory element (CRE) confirm this site to be essential for the progesterone-mediated effects. In summary, our results demonstrate that progesterone regulates CRH gene transcription via a CRE in the CRH promoter and that PR-A and PR-B exhibit different actions in the regulation of CRH gene expression. PMID- 15112059 TI - A general method for the analysis of random bisubstrate enzyme mechanisms. AB - In the present communication, a general method for the kinetic analysis of random bisubstrate mechanisms is described. The method comprises a stepwise application of the following kinetic and ligand-binding experiments: determination of steady state kinetic constants, product inhibition patterns, maximum rate relationships, application of alternate substrates, application of dead-end inhibitors, direct binding of substrates, kinetic isotope effects, and isotope exchange studies. This general method was applied to a practical example: a yeast alcohol dehydrogenase-catalyzed oxidation of 2-propanol by NAD(+) at pH 7.0, 25 degrees C. It was found that this fully reversible reaction proceeds by a steady-state random Bi-Bi mechanism, whereby both dead-end complexes are formed. PMID- 15112060 TI - Strain improvement for fermentation and biocatalysis processes by genetic engineering technology. AB - Twenty years ago, the first complete gene cluster encoding the actinorhodin biosynthetic pathway was cloned and characterized. Subsequently, the gene clusters encoding the biosynthetic pathways for many antibiotics were isolated. In the past decade, breakthroughs in technology brought that generation of rationally designed or new hybrid metabolites to fruition. Now, the development of high-throughput DNA sequencing and DNA microarray techniques enables researchers to identify the regulatory mechanisms for the overproduction of secondary metabolites and to monitor gene expression during the fermentation cycle, accelerating the rational application of metabolic pathway engineering. How are the new tools of biotechnology currently being applied to improve the production of secondary metabolites? Where will this progress lead us tomorrow? The use of whole cells or partially purified enzymes as catalysts has been increased significantly for chemical synthesis in pharmaceutical and fine chemical industries. The development of PCR technologies for protein engineering and DNA shuffling is leading to the generation of new enzymes with increased stability to a wide range of pHs, temperatures and solvents and with increased substrate specificity, reaction rate and enantioselectivity. Where will this emerging technology lead us in the twenty-first century? PMID- 15112061 TI - Antibacterial effects of knotwood extractives on paper mill bacteria. AB - Hydrophilic knotwood extracts from 18 wood species were assessed in disc diffusion and liquid culture tests for antibacterial effects against three species of paper mill bacteria. The Pinus sylvestris, P. resinosa, P. contorta, and P. banksiana extracts decreased or inhibited bacterial growth. The susceptibility order was P. sylvestris > P. resinosa > P. contorta > P. banksiana, correlating with the concentrations of pinosylvin and pinosylvin monomethyl ether in these wood species. Also, Pseudotsuga menziesii and Thuja occidentalis extracts had a small inhibitory effect. The Gram-positive Bacillus coagulans was more susceptible to the extracts than the Gram-negative Burkholderia multivorans and Alcaligenes xylosoxydans. The main components in the Pinus knotwood extracts were pinosylvin monomethyl ether and pinosylvin, suggesting these to be the active components. Therefore, pure pinosylvin, pinosylvin monomethyl ether, and dihydro-pinosylvin monomethyl ether were also tested. All compounds showed antibacterial effects. However, higher concentrations were needed for these pure compounds than for the knotwood extracts. Pinosylvin had stronger antibacterial effects than pinosylvin monomethyl ether. This work shows that knotwood extracts, especially from Pinus species, have a potential for use as natural biocides in papermaking. PMID- 15112062 TI - Rapid diagnosis of Staphylococcus aureus bacteremia using S. aureus PNA FISH. AB - In the study presented here, the performance of the S. aureus PNA FISH assay was evaluated using 285 blood cultures (from 104 patients) that had gram-positive cocci resembling staphylococci on Gram stain. The new molecular test is based on a fluorescence in situ hybridization assay using peptide nucleic acid probes targeting Staphylococcus aureus 16S rRNA and is designed for the rapid identification of Staphylococcus aureus directly from positive blood cultures. The sensitivity, specificity, and positive and negative predictive values of the S. aureus PNA FISH for the rapid identification of Staphylococcus aureus directly from positive blood culture bottles were 100, 99.4, 99.2 and 100%, respectively. PMID- 15112063 TI - Resistance of gram-negative non-fermentative bacilli causing bloodstream infection, Vienna, 1996-2003. PMID- 15112064 TI - Rapid detection of Mycoplasma pneumoniae IgM antibodies in pediatric patients using ImmunoCard Mycoplasma compared to conventional enzyme immunoassays. PMID- 15112066 TI - Metabolically deficient methicillin-resistant Staphylococcus aureus as cause of chronic post-thoracotomy sternal wound infection. PMID- 15112065 TI - Blastocystis hominis as a cause of hypoalbuminemia and anasarca. AB - The protozoan Blastocystis hominis has been considered nonpathogenic, but this classification has come under scrutiny in light of reports in the medical literature indicating it could be the cause of intestinal disorders and, in one case, hypoalbuminemia. Reported here is a severe case of infection with B. hominis that caused acute gastroenteritis with prolonged diarrhea, hypoalbuminemia and anasarca. The diagnosis was based on the parasitological finding, since no other pathological evidence was found. The patient responded favorably to treatment with metronidazole for 10 days. This case supports the idea that B. hominis should be considered as a cause of opportunistic infection in debilitated patients despite the controversy surrounding its pathogenicity. PMID- 15112067 TI - Four cases of necrotizing fasciitis caused by Klebsiella species. AB - Presented here are four cases of necrotizing fasciitis caused by Klebsiella spp. that were treated at one hospital over a 2-year period. Klebsiella necrotizing fasciitis can occur via direct inoculation, local trauma or, more commonly, hematogenous spread from other septic foci. Early, aggressive, surgical debridement and appropriate antimicrobial treatment are the cornerstones of treatment for this condition. Necrotizing fasciitis due to Klebsiella spp. is unique in that it is commonly associated with multiple septic foci. While liver abscesses and endogenous endophthalmitis are better-known associations of disseminated Klebsiella infection, necrotizing fasciitis is increasingly recognized as one of the manifestations of this syndrome. When treating Klebsiella necrotizing fasciitis, awareness of the potential for multiorgan involvement should prompt a thorough search for associated foci of infection. PMID- 15112068 TI - Severe peritonitis due to Balantidium coli acquired in France. AB - The case reported here concerns an alcoholic pork-butcher who presented with severe colitis with peritonitis, caused by the only ciliate protozoan capable of infecting humans, Balantidium coli. This parasite is common in a variety of domestic and wild mammals, mainly pigs; however, its prevalence rate in humans is very low--particularly in industrialised, northern countries, including France. The infection is most frequently acquired by ingesting food or water contaminated by pig faeces, and it may be asymptomatic or may cause acute diarrhoea. Specific antibiotic treatment is efficacious, and it is important to consider the risk of this parasitic disease in susceptible patients presenting with bloody diarrhoea. PMID- 15112069 TI - Management of hepatitis B in patients coinfected with the human immunodeficiency virus. AB - The human immunodeficiency virus (HIV) and the hepatitis B virus share common routes of transmission, and hence, coinfection with these two viruses is common. Chronic hepatitis B does not influence the progression of HIV disease or the response to highly active antiretroviral therapy. It is clear, however, that HIV infection does impact the course of hepatitis B, as higher rates of chronic carriage, lower seroconversion rates, and accelerated progression towards cirrhosis have been observed. Vaccination against hepatitis B is less effective in HIV-infected individuals. Coinfected subjects have a poor response to interferon therapy. Lamivudine is more effective in coinfected subjects but must not be used as monotherapy because of the risk of resistance developing. Combination therapy with lamivudine and tenofovir has shown promise and is currently being investigated in clinical trials, while new drugs and other combinations are in development. PMID- 15112070 TI - Clinical factors associated with fluconazole resistance and short-term survival in patients with Candida bloodstream infection. AB - In a 1-year national surveillance program of Candida bloodstream infections in Japan, clinical factors predicting fluconazole resistance and survival of the patients were analyzed. Blood isolates and complete clinical histories were obtained from 326 patients. Fluconazole-resistant isolates were found in 15 (4.6%) of the cases. Univariate analysis of the demographic and clinical factors associated with fluconazole resistance revealed that age, hematologic malignancy, neutropenia, and immunosuppression were of statistical significance. A multiple logistic regression model showed that only hematologic malignancy as the underlying disease (odds ratio, 6.6; 95% confidence interval, 1.6-26.9; P=0.009) was independently associated with resistance. In 242 cases in which data regarding management and prognosis were available, the 30-day survival rate was 68.4%. In the univariate analysis of factors predicting survival, a significant association was found for Candida species, age of the patient, neutropenia, recent abdominal surgery, removal of the central venous catheter, and use of appropriate antifungal therapy. In the multivariate analysis, removal of the central venous catheter (odds ratio, 6.0; 95% confidence interval, 2.2-16.1; P<0.001) and the use of appropriate therapy (odds ratio, 2.1; 95% confidence interval, 1.1-4.1; P=0.03) were independent factors significantly associated with survival after the diagnosis of Candida bloodstream infection. PMID- 15112071 TI - Relationship between glycopeptide use and decreased susceptibility to teicoplanin in isolates of coagulase-negative staphylococci. AB - To investigate the relationship between glycopeptide use and decreased susceptibility to teicoplanin in coagulase-negative staphylococci (CNS) isolates, data on teicoplanin susceptibility and glycopeptide use from existing microbiology laboratory and pharmacy databases were collected for the period between July 2000 and March 2001. Pooled data for the entire study period were first used to analyse associations. Univariate analysis showed that the incidence of CNS with decreased susceptibility to teicoplanin was significantly correlated with the use of glycopetides, particularly with vancomycin use. This association was confirmed by multivariate analysis. This study suggests that variations in antimicrobial resistance are related to variations in antimicrobial use in the model of CNS with decreased susceptibility to teicoplanin, thus confirming the usefulness of restricting antimicrobial prescribing as a means of controlling resistance. PMID- 15112072 TI - Evaluation of a disk diffusion method with cefoxitin (30 microg) for detection of methicillin-resistant Staphylococcus aureus. AB - The emergence of heterogeneous populations of methicillin-resistant Staphylococcus aureus (MRSA) causes major problems in routine screening for MRSA. In heterogeneous MRSA populations, a proportion of bacterial cells show low-level resistance to oxacillin, with minimal inhibitory concentrations (MICs) of oxacillin ranging between 1 and 100 mg/l, while in homogeneous MRSA populations, the MIC of oxacillin for all cells is >100 mg/l. Routine oxacillin disk diffusion tests often fail to detect heterogeneous MRSA populations. In the present study, a recently proposed disk diffusion method that employs a cephamycin antibiotic (cefoxitin 30 microg; BD Sensi-disc, Becton Dickinson, Germany) was evaluated using 155 clinical isolates of S. aureus (73 mecA positive and 82 mecA negative). The results were compared with those of other MRSA screening techniques: a disk diffusion test with oxacillin 1 microg and cefoxitin 30 microg (BD Sensi-disc; Becton Dickinson), an MRSA latex agglutination test (Denka Seiken, Japan), and an oxacillin screen agar test (6 microg/ml; Becton Dickinson). Detection of the mecA gene by polymerase chain reaction was considered the gold standard. The performances of the different methods were determined and compared. The results showed that the cefoxitin disk diffusion test is preferable to the oxacillin disk diffusion method for routine screening to detect MRSA. PMID- 15112073 TI - Leprosy and AIDS: two cases of increasing inflammatory reactions at the start of highly active antiretroviral therapy. AB - Reported here are the cases of two HIV-positive patients with skin lesions suggestive of leprosy, based on clinical and pathological analysis, which worsened during the few weeks following initiation of highly active antiretroviral therapy. The lesions improved after a few weeks of multidrug therapy for leprosy. Mycobacterium leprae was confirmed by polymerase chain reaction analysis of blood in case 1 and of a biopsy sample in case 2. Neither Mycobacterium avium complex nucleic acid, which is usually associated with immune restoration syndrome, nor mycobacterial cutaneous manifestations were detected in either case. PMID- 15112074 TI - Current trends in the molecular diagnosis of infective endocarditis. AB - The introduction of molecular methods into biomedical science, particularly nucleic acid amplification techniques including the polymerase chain reaction, has significantly improved the diagnosis of several diseases. Likewise, the adoption of such molecular techniques to aid in the detection and identification of causal organisms in patients with infective endocarditis (IE) has been particularly beneficial in cases of difficult, atypical or culture-negative IE. Several different molecular approaches have been suggested for the diagnosis of IE, including variations in the type of cardiological specimens examined, nucleic acid extraction, gene target and molecular platform, each presenting their own advantages and disadvantages. This review examines the molecular approach to the detection and identification of causal agents of IE and provides details and a discussion of the application of such methods, particularly those implemented over the last 7 years. PMID- 15112075 TI - Double-strand hydrolysis of DNA by a magnesium(II) complex with diethylenetriamine. AB - The development of artificial nucleases that hydrolyze DNA or RNA is of great interest in molecular biology, biotechnology, and medicine. We now report that a magnesium(II) complex of diethylenetriamine (Mg-dien) can effectively promote the double-stranded cleavage of plasmid DNA and the dideoxynucleotide dApdA under physiological conditions of pH and temperature. Experiments performed in the presence of hydrogen peroxide, radical scavengers, or under rigorously anaerobic conditions indicate that DNA cleavage mediated by Mg-dien occurs via a hydrolytic path. Mg-dien efficiently hydrolyzes supercoiled pBR322 DNA and the pseudo-first order rate constant at 37 degrees C and pH 8.0 is estimated to be 1.60 h(-1). The dinucleotide dApdA hydrolysis, with Mg-dien at 170 microM, shows a rate enhancement factor of ca. 5 x 10(8). 1H and 31P(1H) NMR studies show that Mg-dien effectively hydrolyzes 5'-dAMP to give deoxyadenosine and inorganic phosphate. While Mg2+ has been found at the catalytic sites of many natural nucleases, Mg dien appears to be the first synthetic Mg2+-containing system capable of hydrolyzing dideoxynucleotides and DNA and thus may provide a simple model system to assist mechanistic studies of naturally occurring nucleases. PMID- 15112076 TI - Comment on "Evaluation of standard nucleotomy after Love for lumbar disc herniation: results of follow-up studies after more than 10 years," by Y. Saruhashi et al. PMID- 15112077 TI - Use of cervical stand-alone cages. PMID- 15112079 TI - Frugivory in polychrotid lizards: effects of body size. AB - As more data have become available on lizard diets in the past few decades, researchers have stressed the importance of lizards as pollinators and seed dispersers. Whereas large body size has been traditionally put forward as a major biological factor "allowing" herbivory and frugivory in lizards, a recent review of frugivory and seed dispersal by lizards showed that frugivory might be considered to be a typical island phenomenon, independent of body size. Here we show that frugivory is correlated with lizard body size among a group of syntopic Anolis species in Jamaica, with larger species eating more fruit. Additionally, the size of the fruits consumed is significantly related to lizard body size. Multiple regression analyses show that this is largely a pure body size effect as head shape or residual bite force are uncorrelated to overall fruit size. Moreover, we demonstrate that among polychrotid (Anolis-like) lizards in general, those that consume fruit are on average larger than those that do not. Lizards from the mainland were not significantly different in body size from island species. We thus suggest that fruit consumption in polychrotid lizards is mediated by large body size whether living on islands or not. PMID- 15112080 TI - Quantitative analysis of Borrelia burgdorferi gene expression in naturally (tick) infected mouse strains. AB - Adaptation of Borrelia burgdorferi in the vector and vertebrate host is mediated by mechanisms that regulate differential expression of outer surface lipoproteins (Osps). In this study, real time PCR was applied to quantify tissue-specific expression of four linear plasmid (lp54)-encoded (ospA, zs7.a36, zs7.a66 zs7.a68) and one circular plasmid (cp26)-encoded (ospC) gene from B. burgdorferi sensu stricto, in a natural setting of tick-infected immunodeficient (C.B-17 SCID) and immunocompetent (BALB/c and AKR/OlaHsd) mice for up to 120 days post-infection (p.i.). Early during infection (day 30 p.i.) high numbers of spirochetes were found in the heart and joint, but not the ear and spleen tissues of disease susceptible SCID mice. In disease-susceptible AKR mice spirochetes colonized the ear and joint tissues, but were undetectable in tissues of disease-resistant BALB/c mice. Later in infection (day 120 p.i.), spirochetes had expanded (approximately 1,000-fold) in all SCID tissues tested but were undetectable in AKR and BALB/c mice. Of the five genes analyzed, only zs7.a36 transcripts were detected in various tissues of all infected mouse strains, though at differing levels, whereas ospC transcripts were only found in tissue specimens of SCID mice. Furthermore, gene expression of ospC and zs7.a36 appears to be differentially regulated in distinct organs of individual mice. In contrast, transcripts for ospA, zs7.a66, and zs7.a68 were not detected in any of the mouse strains, independent of their immune status and/or the severity of their infection/inflammatory responses. Late during infection (day 120 p.i.), transcription of zs7.a36 and ospC was down-regulated in the tissues of SCID mice despite expansion of spirochetes. This type of quantitative analysis may be helpful to further disclose principles of pathogenesis of Lyme borreliosis and to design strategies for its therapeutic treatment. PMID- 15112081 TI - Distribution of choline acetyltransferase and NADPH diaphorase in the spinal cord of the pigeon. AB - Acetylcholine is the neurotransmitter of somatic and autonomic motor systems of the spinal cord. However, there are also intrinsic cholinergic systems which have modulatory functions. Modulatory functions have also been assigned to nitric oxide (NO). Acetylcholine is synthesized by choline acetyltransferase and NO by nitric oxide synthase, which is a NADPH diaphorase. The distribution of both enzymes in the mammalian spinal cord is well known. However, there is a lack of comparative data in avian species. Therefore, the distribution of both enzymes in the spinal cord of the pigeon was studied using histochemical and immunohistochemical methods. Aside from somatic motor neurons and autonomic preganglionic neurons choline acetyltransferase-immunoreactivity was found throughout the spinal cord in lamina III of the superficial dorsal horn and near the central canal. The location of choline acetyltransferase-positive preganglionic neurons in the centrally located column of Terni and the lack of an intermediolateral column typical of the mammalian spinal cord can be confirmed. In lumbosacral segments the axons of centrally located cholinergic neurons crossed to the contralateral side to form a tract in the ventral funiculus, which then innervates the contralateral grey substance. A dense band of NADPH diaphorase staining was found in lamina II and in centrally located neurons of all segments. Part of the centrally located neurons double-labelled for choline acetyltransferase and NADPH diaphorase. In contrast to mammals, preganglionic neurons labelled only weakly for NADPH diaphorase. Altogether, despite the divergent evolution of both classes of vertebrate intrinsic modulatory choline acetyltransferase and NADPH diaphorase systems of birds seem to be largely similar to those of the mammalian spinal cord. PMID- 15112082 TI - Carbonic anhydrase isoform expression and functional role in rodent extraocular muscle. AB - Carbonic anhydrase (CA) accelerates contractile function, particularly in fast twitch skeletal muscles. Since the extraocular muscles are considered to be amongst the fastest skeletal muscles in mammals, this study tested two hypotheses: (1) CA is expressed at higher levels in rat extraocular muscles than in extensor digitorum longus (EDL, a fast limb muscle), and (2) inhibition of CA activity increases twitch duration and force in the extraocular muscles to a greater extent than in EDL. By real-time quantitative PCR we determined that the expression of CA3 isoform, typically high in skeletal muscles, is significantly depressed in extraocular muscles. Message levels for the CA2 and CA4 isoforms were higher in the extraocular muscles, while CA5 expression was equivalent in both muscles. Strong CA activity was demonstrated by histochemistry in frozen EDL muscle sections, in particular along the sarcolemma and in capillaries. By contrast, extraocular muscle had very low sarcolemmal or cytosolic CA activity. CA inhibition with 6-ethoxyzolamide (ETZ) reversibly increased twitch duration and force in EDL muscle bundles. In the extraocular muscles, ETZ did not alter twitch kinetics. Based on these results, we reject our initial hypotheses and conclude that CA does not influence the fast contractile kinetics characteristic of the extraocular muscles. PMID- 15112083 TI - Skin reactions due to low molecular weight heparin in pregnancy: a strategic dilemma. AB - INTRODUCTION: Heparin-induced allergic reactions may cause problems if heparin administration is needed for thrombo-embolic disease in pregnancy. CASE REPORTS: We report two cases of hypersensitivity to low molecular weight heparin (LMWH) in pregnancy. DISCUSSION: Alternative methods and new antithrombotic agents are discussed. PMID- 15112084 TI - Unstable versus stable uncemented femoral stems: a radiological study of periprosthetic bone changes in two types of uncemented stems with different concepts of fixation. AB - INTRODUCTION: Uncemented stems in total hip arthroplasty (THA) are used increasingly often because they are believed to offer a reliable long-term fixation. However, periprosthetic bone remodelling has been a worrying issue. A proximal demineralization has been noted in femurs with well-fixed stems, and it has been explained as by-passing of mechanical forces along the fixed implant (stress-shielding). Aseptic loosening has been a major problem in several uncemented series with earlier designs. The objective for this study was to investigate how the host bone adapts to a loose stem compared with a well-fixed stem after a long time. MATERIALS AND METHODS: An investigation with dual-energy X-ray absorptiometry (DEXA), scintimetry and radiological assessment was carried out in 20 patients 8 years after a THA for arthrosis with two different uncemented stems. Ten patients received a stem coated with polytetrafluoroethylene (Anaform); all prostheses showed migration and were considered unstable. Ten patients received a hydroxyapatite-coated stem (Bi Metric); no prosthesis migrated. RESULTS: Different remodelling patterns were seen. In the unstable group, the periprosthetic bone mineral density (BMD) was significantly reduced along the entire stem, while in the stable group only proximal bone loss was seen. The scintigraphic uptake was increased under the stem tip in both groups, and among unstable stems uptake was also increased in the calcar region. CONCLUSION: The assessment of periprosthetic bone remodelling after uncemented THA with long-term observation shows a different host-bone response in stable versus unstable femoral implants. Prior to a femoral revision, measurement of the BMD could be beneficial; it may guide the surgeon when deciding which surgical technique to use. PMID- 15112085 TI - Transient osteoporosis of the navicular bone in a runner. PMID- 15112087 TI - [Neurological manifestations of rheumatic disorders]. PMID- 15112088 TI - [Neurological aspects of systemic rheumatological disorders]. AB - There are a number of common aspects between rheumatology and neurology. First, many systemic rheumatic diseases also affect the central and peripheral nervous system as well as muscle causing characteristic symptom complexes. Second, there are similarities between neurological and rheumatological autoimmune diseases in terms of underlying pathomechanisms. Here the most important neurological aspects of rheumatological disorders including their diagnosis and therapy are highlighted. PMID- 15112089 TI - [Psychosomatics in rheumatology]. AB - Psychosocial factors influence the course and the outcome of chronic somatic diseases. This is also valid for rheumatic diseases like rheumatoid arthritis, spondyloarthropathies, systemic collagen vascular diseases, and fibromyalgia syndrome. The article summarises the evidence-based findings and it illustrates possibilities of psychosomatic treatment in rheumatic diseases by means of three case reports. PMID- 15112090 TI - [Psychiatric disorders in rheumatic diseases, as exemplified by systemic lupus erythematosus (SLE)]. AB - Among the rheumatic diseases, systemic lupus erythematosus (SLE) reveals the highest frequency of central nervous symptoms. Psychiatric abnormalities are present in over 90% of the patients, if very mild cognitive impairment is included. The psychiatric syndromes listed by the American College of Rheumatologists encompass cognitive syndromes, including the acute confusional state, anxiety disorders, depressive symptomatology and psychosis. Associations between cognitive impairment and demographic and disease variables, i. e. disease severity, have not been clearly identified so far. The same holds true for the evolution of depressive symptoms. Reactive depression in coping with a chronic disease is, however, a psychologically plausible factor in addition to specific cerebral lesions. Therapeutic interventions in SLE should consist of a combined pharmacological and nonpharmacological treatment. Corticosteroid drugs, however, may be effective in alleviating mild psychiatric symptoms as well. PMID- 15112091 TI - [Merger in rheumatology]. PMID- 15112092 TI - [Structural change in the DGRh]. PMID- 15112093 TI - [The infected rheumatoid forefoot--how much surgery is advisable?]. AB - A biomechanically altered rheumatic forefoot with a resulting shoe conflict and an impaired immune system due to underlying disease and medical treatment are often cause for infection. Often infected bursae perforate into the corresponding joints and may consequently lead to severe infectious states of the foot. The multiplicity of possible biomechanical adjustments makes it difficult to develop an undisputed concept. Conservative versus operative therapeutic options are possible and may be individually applied. It should be questioned, if the same general rules in septic joint surgery also apply to the rheumatoid forefoot. This retrospective clinical study will evaluate a concept for possible operative treatment of the infected rheumatoid forefoot in selected cases. Our conception advocates operative procedures on a patient adapted basis in order to prevent recurrent infections. Suggestions for operative procedures are given. PMID- 15112094 TI - [Navigated implantation of total knee endoprostheses in secondary knee osteoarthritis of rheumatoid arthritis patients as compared to conventional technique]. AB - INTRODUCTION: The operative treatment of a secondary gonarthrosis due to RA claims high quality in soft tissue balancing and accurate alignment in total knee arthroplasty (TKA) which are essential for good long-term results. The efficiency of an imageless computer-assisted implantation in TKA was evaluated and compared with conventional technique. METHOD: The authors implanted each 40 TKA either using the imageless computer-assisted or classical surgeon-controlled technique. The quality of implantation was studied on postoperative long leg coronal and lateral x-rays. RESULTS: A postoperative leg axis between 3 degrees varus and 3 degrees valgus was obtained in 37 patients (92.5%) in the study group and 30 patients (75%) in the control group. Complications influencing the clinical outcome did not occur. CONCLUSION: The use of the imageless Vector-Vision navigation system provides the patient a good chance for longterm survival. Especially the ligament balancing tool appears to be useful. Cutting errors can be detected and intraoperatively corrected. PMID- 15112095 TI - [Evidence-based use of methotrexate in children with rheumatic disorders. Consensus statement of the Working Group for Children and Adolescents with Rheumatic Diseases in Germany (AGKJR) and the Working Group Pediatric Rheumatology Austria]. AB - Juvenile idiopathic arthritis (JIA) is the most common diagnosis in children and adolescents with rheumatic disorders. In many children and adolescents, JIA is successfully treated with nonsteroidal antiinflammatory drugs (NSAR) and physiotherapy. Still, in a significant number of cases the disease is resistant to this therapy and treatment with "second line" disease modifying antirheumatic drugs (DMARDs) is required. Methotrexate (MTX) is frequently referred to as "first choice second line agent" for the treatment of JIA. However, there are considerable differences among pediatric rheumatologists on how and when to use MTX. To increase drug safety, the Working Group for Children and Adolescents with Rheumatic Diseases in Germany (AGKJR) and the Working Group Pediatric Rheumatology Austria have initiated the formulation of evidence-based recommendations. Evidence is based on consensus expert meetings, a MEDLINE search with the key words "Methotrexate" and "juvenile arthritis" limited to age 0-18 years, standard textbooks and review articles, data from the central registry of the German Research Center for Rheumatic Diseases (Deutsches Rheumaforschungszentrum Berlin), experience with MTX in adults with rheumatoid arthritis (RA) and recommendations of the German Society of Rheumatology (DGRh). Based on these data, evidence and recommendations are graded and evidence-based recommendations for the use of MTX in children and adolescents with rheumatic disease are presented. PMID- 15112096 TI - [Rheumatology: Integration into student training--the RISA- Study. Results of a survey exploring the scale of education and training in rheumatology at German universities]. AB - During the Bone and Joint Decade 2000-2010, national and international rheumatological societies campaign for improvements of undergraduate training in the field of musculoskeletal diseases. In 2002, the Committee for Undergraduate Training of the German Society for Rheumatology (DGRh) performed a survey in order to estimate the extent of rheumatological undergraduate training at German universities. A questionnaire was sent to all university hospitals for internal medicine and for orthopaedics. The items of the survey covered the qualification of the teachers, the number and the time frame of lessons and courses, as well as the items of the curriculum in rheumatology. With 95% of the universities responding, a broad variation of the quality of rheumatological training became obvious, ranging from the absence of competent teaching to a comprehensive curriculum covering theory and practical training. Only a minority of universities fulfilled the recommendations of the DGRh for undergraduate education. The most substantial deficits are found in practical training, caused by insufficiently small numbers of patients seen by the medical students, and by the short duration of the practical courses. The insights described here should lead to the adaptation of the level of education in the field of rheumatology to the demands of a society with growing musculoskeletal problems. PMID- 15112097 TI - [Hans Reiter and the oculo-urethro-synovial syndrome. 2. Reiter and "his" disease -- the hunt for the chimera]. PMID- 15112098 TI - [Progress and promise in rheumatology, measures of success. 19-21 March 2004 in Barcelona]. PMID- 15112101 TI - Specializations for aerial hawking in the echolocation system of Molossus molossus (Molossidae, Chiroptera). AB - While searching for prey, Molossus molossus broadcasts narrow-band calls of 11.42 ms organized in pairs of pulses that alternate in frequency. The first signal of the pair is at 34.5 kHz, the second at 39.6 kHz. Pairs of calls with changing frequencies were only emitted when the interpulse intervals were below 200 ms. Maximum duty cycles during search phase are close to 20%. Frequency alternation of search calls is interpreted as a mechanism for increasing duty cycle and thus the temporal continuity of scanning, as well as increasing the detection range. A neurophysiological correlate for the processing of search calls was found in the inferior colliculus. 64% of neurons respond to frequencies in the 30- to 40-kHz range and only in this frequency range were closed tuning curves found for levels below 40 dB SPL. In addition, 15% of the neurons have double-tuned frequency threshold curves with best thresholds at 34 and 39 kHz. Differing from observations in other bats, approach calls of M. molossus are longer and of higher frequencies than search calls. Close to the roost, the call frequency is increased to 45.0-49.8 kHz and, in addition, extremely broadband signals are emitted. This demonstrates high plasticity of call design. PMID- 15112102 TI - Fine mapping of a seizure susceptibility locus on mouse Chromosome 1: nomination of Kcnj10 as a causative gene. AB - Previous quantitative trait loci (QTL) mapping studies document that the distal region of mouse Chromosome (Chr) 1 contains a gene(s) that is in large part responsible for the difference in seizure susceptibility between C57BL/6 (B6) (relatively seizure-resistant) and DBA/2 (D2) (relatively seizure-sensitive) mice. We now confirm this seizure-related QTL ( Szs1) using reciprocal, interval specific congenic strains and map it to a 6.6-Mb segment between Pbx1 and D1Mit150. Haplotype conservation between strains within this segment suggests that Szs1 may be localized more precisely to a 4.1-Mb critical interval between Fcgr3 and D1Mit150. We compared the coding region sequences of candidate genes between B6 and D2 mice using RT-PCR, amplification from genomic DNA, and database searching and discovered 12 brain-expressed genes with SNPs that predict a protein amino acid variation. Of these, the most compelling seizure susceptibility candidate is Kcnj10. A survey of the Kcnj10 SNP among other inbred mouse strains revealed a significant effect on seizure sensitivity such that most strains possessing a haplotype containing the B6 variant of Kcnj10 have higher seizure thresholds than those strains possessing the D2 variant. The unique role of inward-rectifying potassium ion channels in membrane physiology coupled with previous strong association between ion channel gene mutations and seizure phenotypes puts even greater focus on Kcnj10 in the present model. In summary, we confirmed a seizure-related QTL of large effect on mouse Chr 1 and mapped it to a finely delimited region. The critical interval contains several candidate genes, one of which, Kcnj10, exhibits a potentially important polymorphism with regard to fundamental aspects of seizure susceptibility. PMID- 15112103 TI - Comparative physical maps of the human and mouse Meckel syndrome critical regions. AB - Meckel syndrome (MKS-OMIM 24900) is an autosomal recessive disease characterized by cystic kidneys, occipital encephalocele, polydactyly, and fibrotic changes of the liver, typically resulting in postnatal death. A Meckel syndrome critical region (MKS1) maps to human Chromosome (Chr) 17, in a region of homology to mouse Chr 11. Here we report the comparison of human Chr 17q23 with mouse Chr 11. We have generated physical maps of the human and mouse MKS1 critical regions. Additionally, we have created a transcript map of the MKS1 critical region in both species. By comparing these physical maps, we observe a high degree of similarity in gene order in the human and mouse Meckel syndrome critical regions. We have also examined the expression patterns of genes in the MKS1 region to assess their potential as MKS1 candidates. Finally, we have analyzed genes present in the other Meckel syndrome critical regions, MKS2 and MKS3, to determine whether any of the candidate genes for the three MKS loci have similar gene functions or are members of a common biological pathway. PMID- 15112104 TI - Gene content of the 750-kb critical region for mouse embryonic ectoderm lethal tcl-w5. AB - Mice homozygous for the t(w5) allele arrest at gastrulation from defects associated with embryonic ectoderm development. The mutated gene has been genetically closely linked to the H-2K locus in the mouse MHC region, flanked by markers H-2Pb and D17Mit147. Aiming at the positional cloning of the mutated gene, we constructed a BAC contig spanning about 1 Mb of the genomic region. On the basis of our mapping and sequencing analysis of the BACs combined with public genome data, EST database searches, and gene prediction programs, we delimit the 1.06 cM of the t(w5) critical region to 750 kb, and infer 36 genes (1/20 kb) encoded in the interval. All of the 33 genes tested were confirmed as expressed in embryonic tissues by RT-PCR analyses, and in many cases by EST expression profiles as well. Thus, this highly gene-rich region is essentially totally transcribed during early development and provides priority candidates to be screened for the t(w5) embryonic lesion. PMID- 15112105 TI - New retinal light damage QTL in mice with the light-sensitive RPE65 LEU variant. AB - The purpose of this study was to determine the QTL that influence acute, light induced retinal degeneration differences between the BALB/cByJ and 129S1/SvImJ mouse strains. Five- to 6-week-old F(2) progeny of an intercross between the two strains were exposed to 15,000 LUX of white light for 1 h after their pupils were dilated, placed in the dark for 16 h, and kept for 10-12 days in dim cyclic light before retinal rhodopsin was measured spectrophotometrically. This was used as the quantitative trait for retinal degeneration. Neither gender nor pigmentation had a significant influence on the amount of rhodopsin after light exposure in the F(2) progeny. For genetic study, DNAs of the 27-36 F(2) progeny with the highest and 27-36 F(2) with the lowest levels of rhodopsin after light exposure were genotyped with 71 dinucleotide repeat markers spanning the genome. Any marker with a 95% probability of being associated with phenotype was tested in all 289 F(2) progeny. Data were analyzed with Map Manager QTX. Significant QTL were found on mouse Chrs 1 and 4, and suggestive QTL on Chrs 6 and 2. The four QTL together equal an estimated 78% of the total genetic effect, and each of the QTL represents a gene with BALB/c susceptible alleles. The Chr 6 QTL is in the same region as a highly significant age-related retinal degeneration QTL found previously. Identification of these QTL is a first step toward identifying the modifier genes/alleles they represent, and identification of the modifiers may provide important information for human retinal diseases that are accelerated by light exposure. PMID- 15112106 TI - Analysis of imprinted murine Peg3 locus in transgenic mice. AB - Peg3 is an imprinted gene exclusively expressed from the paternal allele. It encodes a C(2)H(2) type zinc-finger protein and is involved in maternal behavior. It is important for TNF-NFkB signaling and p53-mediated apoptosis. To investigate the imprinting mechanism and gene expression of Peg3 and its neighboring gene(s), we used a 120 kb Peg3-containing BAC clone to generate transgenic mice. The BAC clone contains 20 kb of 5' and 80 kb of 3' flanking DNA, and we obtained three transgenic lines. In one of the lines harboring one copy of the transgene, Peg3 was imprinted properly. In the other two lines, Peg3 was expressed upon both maternal and paternal transmission. Imprinted expression was linked to the differential methylation of a region (DMR) upstream of the Peg3 gene. A second, maternally expressed gene, Zim1, present on the transgene was expressed irrespective of parental inheritance in all lines. These data suggest that, similar to other imprinted genes within domains, Peg3 and Zim1 are regulated by one or more elements lying at a distance from the genes. The imprinting of Peg3 seen in one line may reflect the presence of a responder sequence. Concerning the expression of the Peg3 transgene, we detected appropriate expression in the adult brain. However, this was not sufficient to rescue the maternal behavior phenotype seen in Peg3 deficient animals. PMID- 15112107 TI - A comparative analysis of the pig, mouse, and human PCDHX genes. AB - Protocadherin X and Y (PCDHX/Y) represent a pair of homologous genes located on the human sex chromosomes that are primarily expressed in the brain. PCDHY emerged as a result of a duplicative transposition from the X Chromosome (Chr) and is present on the Y only in hominids. Previous zoo-blot analysis suggested the existence of PCDHX orthologs on the X Chr of several mammalian species. This paper reports the cloning and characterization of porcine and murine Pcdhx. Pig Pcdhx cDNA was obtained by a combination of RT-PCR, SMART-RACE, and genomic sequencing and exhibits 88% identity to human PCDHX; FISH analysis indicated that porcine Pcdhx maps to Xq. Mouse Pcdhx cDNA was assembled by RT-PCR and database analysis and is 84% identical to the human gene. Some degree of alternative splicing was detected in pig Pcdhx, but not to the extent previously described in humans. Both murine and porcine Pcdhx mRNA were detected in all tissues studied. Cloning of 2.5 kb of genomic sequence upstream of the most 5' exon of porcine Pcdhx allowed a comparative analysis with murine and human sequences in order to define potential promoter elements. All exons present in mouse and pig transcripts were found to have homologous sequences in human DNA. Not all of these exons are represented in human transcripts, indicating differential evolution and usage. The increased complexity in post-transcriptional processing and restriction of expression of the human genes primarily to central nervous system tissue as compared with pig and mouse suggests that PCDHX/Y is potentially a good candidate to account for human-specific features of the CNS. PMID- 15112108 TI - The rat Ruby ( R) locus is Rab38: identical mutations in Fawn-hooded and Tester Moriyama rats derived from an ancestral Long Evans rat sub-strain. AB - Hermansky-Pudlak syndrome (HPS) is a group of rare, recessive disorders in which oculocutaneous albinism, progressive pulmonary fibrosis, bleeding diathesis, and other abnormalities result from defective biogenesis of multiple cytoplasmic organelles. Seven different HPS genes are known in humans; in mouse, at least 16 loci are associated with HPS-like mutant phenotypes. In the rat, only two HPS models are known, Fawn-hooded (FH) and Tester Moriyama (TM), non-complementing strains in which HPS-like hypopigmentation and platelet storage pool deficiency result from a mutation of the Ruby (red eyed dilution; R) locus on Chromosome (Chr) 1. We have identified the R locus as the Rab38 gene, establishing that rat R is homologous to mouse chocolate ( cht). Further, we show that FH and TM rats have identical Rab38 Met1Ile mutations, occurring on an identical Chr 1 marker allele haplotype, indicating that these two strains derive from a common ancestor. This ancestor appears to have been a sub-strain of the outbred Long Evans (LE) strain, and several modern LE sub-strains carry the Rab38 Met1Ile R mutation on the same Chr 1 marker haplotype. These findings have significant implications for the many past and ongoing studies that involve the FH and LE derivative rat strains. Hermansky-Pudlak syndrome (HPS; MIM 203300) is a group of autosomal recessive diseases in which oculocutaneous albinism (OCA), progressive and fatal pulmonary fibrosis, and bleeding diathesis due to platelet storage pool deficiency result from defects in the biogenesis of specific cytoplasmic organelles and granules: melanosomes, lysosomes, and platelet dense granules. In humans, seven different HPS genes are known. In the mouse, at least 16 loci associated with HPS-like mutant phenotypes are known, seven of which are homologous to the human HPS loci. PMID- 15112109 TI - Characterization of 298 ESTs from porcine back fat tissue and their assignment to the SSRH radiation hybrid map. AB - Following several criteria, we collected, clustered, and functionally categorized 653 expressed sequence tags (ESTs) of 5' ends from porcine back fat libraries from the >15,000 porcine ESTs collected to date. By searching the LocusLink and Mapviewer database, we knew the positions of these 653 ESTs on human chromosomes (HSAs). Sus scrofa radiation hybrid (SSRH) mapping revealed that 298 porcine EST clusters out of 653 were localized near microsatellite (MS) markers. Among these EST clusters, we could assign 182 to their porcine chromosomes (SSCs) on the SSRH map. PMID- 15112110 TI - Intragenic deletion in the gene encoding L-gulonolactone oxidase causes vitamin C deficiency in pigs. AB - The absence of L-ascorbic acid (L-AA, or AA) synthesis in scurvy-prone organisms, including humans, other primates, guinea pigs, and flying mammals, was traced to the lack of L-gulonolactone oxidase (GULO) activity. GULO is a microsomal enzyme that catalyzes the terminal step in the biosynthesis of L-AA. Clinical cases of scurvy were described in a family of Danish pigs. This trait is controlled by a single autosomal recessive allele designated od (osteogenic disorder). Here we demonstrate that the absence of GULO activity and the associated vitamin C deficiency in od/od pigs is due to the occurrence of a 4.2-kbp deletion in the GULO gene. This deletion includes 77 bp of exon VIII, 398 bp of intron 7 and 3.7 kbp of intron 8, which leads to a frame shift. The mutant protein is truncated to 356 amino acids, but only the first 236 amino acids are identical to the wild type GULO protein. In addition, the od allele seems to be less expressed in deficient and heterozygous pigs compared with the normal allele in heterozygous and wild-type animals as determined by ribonuclease protection assay. We also developed a DNA-based test for the diagnosis of the deficient allele. However, we failed to identify the mutated allele in other pig populations. PMID- 15112111 TI - FDG-PET in monitoring therapy of breast cancer. AB - Fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) has been used successfully for the staging and re-staging of breast cancer. Another significant indication is the evaluation of therapy response. Only limited data are available on the use of FDG-PET in breast cancer after radiation therapy. The same holds true for chemotherapy. Only the therapy response in locally advanced breast cancer after chemotherapy has been investigated thoroughly. Histopathological response could be predicted with an accuracy of 88-91% after the first and second courses of therapy. A quantitative evaluation is, of course, a prerequisite when FDG-PET is used for therapy monitoring. Only a small number of studies have focussed on hormone therapy. In this context, a flare phenomenon with increasing standardised uptake values after initiation of tamoxifen therapy has been observed. More prospective multicentre trials will be needed to make FDG PET a powerful tool in monitoring chemotherapy in breast cancer. PMID- 15112112 TI - Pitfalls of magnetic resonance imaging of alar ligament. AB - An observational study of variations in the appearance of the alar ligament on magnetic resonance imaging (MRI) and the normal range of lateral flexion and rotation of the atlas was performed to validate some of the premises underlying the use of MRI for the detection of injuries to the alar ligament. Fifteen healthy volunteers were included. Three sets of coronal proton-density images, and axial T2-weighted images of the craniovertebral junction, were obtained at 0.5 T with the neck in neutral position and laterally flexed (coronal proton density) or rotated (axial T2). Five of the subjects also underwent imaging at 1.5 T. The scans were independently examined twice by two radiologists. The presence of alar ligaments was recorded and a three-point scale used to grade the extent of hyperintensity exhibited by the structures: the ligament were graded as 2 and 3 if, respectively, less or more of its cross-section was hyperintense, whereas grade 1 represented a hypointense ligament. The effect of lateral flexion on image quality was assessed. Concordance analysis of the data were performed before and after dichotomising the data on grading. The atlanto-axial angle and rotation of the atlas were measured. The magnitude of movement to right was normalised to that to the left to give, respectively, the flexion index and the rotation index. The alar ligaments were most reliably seen on coronal proton density scans, with a Maxwell's RE of 0.96 as compared with 0.46 for sagittal images. Flexion of the neck improved definition of the ligaments in only rare instances. Inter-observer disagreement was marked with respect to grading of the ligament on both coronal [composite proportion of agreement (p0)=0.44; 95% confidence intervals: 0.26, 0.64)] and sagittal scans [p0=0.40 (0.19, 0.63)]. Dichotomising the data did not appreciably improve reliability [Maxwell's RE: 0.11 (coronal scans), -0.20 (sagittal scans)]: for ligaments which demonstrated hypertensive areas (grades 2 and 3) there was complete lack of agreement for both coronal [p2=0 (0, 0.25)] and sagittal scans [p2=0 (0-0.30)]. A large response bias was found in the reports of both readers albeit in opposite directions. There was poor concordance between scans obtained at different field strengths [RE (coronal images)=0.25; RE (sagittal images)=0.14). Mean flexion index and mean rotation index were 1.00 (SD 0.03) and 1.01 (SD 0.06), respectively. The MR imaging may not be the investigation of choice for the investigation of subtle injuries to the alar ligament. Whether it can be substituted by kinematic assessment of the occipito-atlanto-axial complex with MRI warrants investigation. PMID- 15112114 TI - Effects of ventral hippocampal long-term potentiation and depression on the gamma band local field potential in anesthetized rats. AB - We examined the effect of long-term potentiation or depression (LTP or LTD) on the local field potential, focusing on the gamma-band (40-100 Hz) power, in the ventral hippocampus CA1 of anesthetized rats. LTP and LTD induction in the CA3 CA1 pathway increased the CA1 spontaneous gamma-band power by around 40 and 80 100 Hz, respectively, while neither changed the evoked levels significantly. These results suggest that the ventral CA1 local field potential can maintain bidirectional plasticity in the steady state for the long term. Given the involvement of synaptic plasticity in learning and memory, the gamma-band power change associated with LTP/LTD may relate to ventral hippocampal functions. The LTP increased the spontaneous power at around 40 Hz of the gamma-band frequency in the ventral CA1, and the LTD did the same at 80-100 Hz. The biphasic increase may distribute the subsequent input appropriately to regulate the relevant synaptic history in the ventral CA1 and anatomically related structures in vivo. PMID- 15112115 TI - Multijoint dynamics and postural stability of the human arm. AB - The goal of this study was to examine how the mechanical properties of the human arm are modulated during isometric force regulation tasks. Specifically, we examined whether the dynamic stability of the limb remained nearly invariant across a range of voluntarily generated endpoint forces and limb postures. Previous single joint studies have demonstrated that dynamic joint stability, as quantified via estimates of the joint damping ratio, is nearly invariant during isometric torque regulation tasks. However, the relevance of these findings to the control of multijoint posture has not been investigated previously. A similar degree of invariance at the multijoint level could suggest a fundamental property of the motor system that could be incorporated into the planning and execution of multijoint tasks. In this work, limb mechanics were quantified using estimates of dynamic endpoint stiffness, which characterizes the relationship between imposed displacements of limb posture and the forces opposing those displacements. Endpoint stiffness was estimated using a two-link robot operating in the horizontal plane at the height of each subject's glenohumeral joint. The robot was used to apply stochastic position perturbations to the arm and to measure the resulting forces. Endpoint stiffness dynamics were estimated nonparametrically and subsequently summarized using inertial, viscous and elastic parameters. We found that in the tasks studied, there was a differential modulation of endpoint elasticity and endpoint viscosity. Elasticity increased nearly linearly with increases in voluntary force generation while viscosity increased nonlinearly. This differential regulation resulted in limb dynamics that had a remarkably consistent damping ratio across all subjects and all tested conditions. These results emphasize the importance of considering the full dynamic response of a limb when investigating multijoint stability, and suggest that a minimal degree of limb stability is maintained over a wide range of force regulation tasks. PMID- 15112116 TI - The effects of increasing memory load on the directional accuracy of pointing movements to remembered targets. AB - The directional accuracy of pointing arm movements to remembered targets in conditions of increasing memory load was investigated using a modified version of the Sternberg's context-recall memory-scanning task. Series of 2, 3 or 4 targets (chosen randomly from a set of 16 targets around a central starting point in 2D space) were presented sequentially, followed by a cue target randomly selected from the series excluding the last one. The subject had to move to the location of the next target in the series. Correct movements were those that ended closer to the instructed target than any other target in the series while all other movements were considered as serial order errors. Increasing memory load resulted in a large decrease in the directional accuracy or equivalently in the directional information transmitted by the motor system. The constant directional error varied with target direction in a systematic fashion reproducing previous results and suggesting the same systematic distortion of the representation of direction in different memory delay tasks. The constant directional error was not altered by increasing memory load, contradicting our hypothesis that it might reflect a cognitive strategy for better remembering spatial locations in conditions of increasing uncertainty. Increasing memory load resulted in a linear increase of mean response time and variable directional error and a non-linear increase in the percentage of serial order errors. Also the percentage of serial order errors for the last presented target in the series was smaller (recency effect). The difference between serial order and directional spatial accuracy is supported by neurophysiological and functional anatomical evidence of working memory subsystems in the prefrontal cortex. PMID- 15112117 TI - Chemosensory additivity in trigeminal chemoreception as reflected by detection of mixtures. AB - A series of experiments probed into the degree of chemosensory detection additivity exhibited by mixtures of ethyl propanoate and heptanoate in terms of their trigeminal detectability via nasal pungency (i.e., irritation) and eye irritation. Nasal pungency was tested in subjects lacking a functional sense of smell (i.e., anosmics) to avoid olfactory biases. First, we built concentration detection functions for each chemical and sensory endpoint. Second, we used the data from the functions to prepare mixtures of the two compounds in complementary proportions, and suitable single-chemical standards, all of which should be equally detectable under a rule of complete additivity, i.e., independence of detection. Third, we compared the experimentally obtained detectability with that expected under such rule. The outcome revealed that, at a low detectability level (but still above chance), the mixtures showed complete additivity for both trigeminal endpoints. At a high detectability level (but below perfect detection), the mixtures showed complete additivity for nasal pungency but less than complete additivity for eye irritation. In the context of previous studies, the results consolidate a picture of higher degree of detection additivity at perithreshold levels in trigeminal than in olfactory chemoreception. The outcome presents another line of evidence suggesting broader chemical tuning in chemesthesis compared to olfaction. PMID- 15112118 TI - Impaired inhibition of a pre-planned response in focal hand dystonia. AB - The hypothesis that focal hand dystonia (FHD) is associated with impaired inhibitory function is supported by a broad base of evidence from studies of neurophysiological function. The purpose of this study was to investigate the inhibition of a pre-planned response in FHD and control subjects. Nine FHD subjects and 11 control subjects performed an anticipated response task, which required them to lift their index finger from a keyboard button to stop the sweep of an indicator at a specified point on a sweep-dial. The affected hands of the FHD subjects were tested, as were the dominant hands of the control subjects. Electromyographic (EMG) data were recorded from the long finger extensors of the tested arm. On 90 out of 220 trials, the sweep indicator would stop prior to the specified point, and subjects were instructed to continue holding the key down when this occurred. The sweep indicator stopped 10 times at each of nine different points around the dial in a randomised order. The accuracy of subjects' performance when stopping the sweep indicator at the specified point, and the proportion of 'stop' trials where they successfully inhibited this pre-planned response, were recorded. The sweep indicator stop time at which each subject had a 50% probability of successfully inhibiting their response (R50) was calculated, as was the probability of extensor EMG (burst) activity occurring during a successfully inhibited response. For FHD subjects, it was found that R50 was significantly lower, and the probability of an extensor EMG burst occurring during a successfully inhibited response was significantly higher, compared to control subjects. These results are discussed with respect to the 'horse-race' model for the inhibition of pre-planned responses. PMID- 15112119 TI - Unifying multisensory signals across time and space. AB - The brain integrates information from multiple sensory modalities and, through this process, generates a coherent and apparently seamless percept of the external world. Although multisensory integration typically binds information that is derived from the same event, when multisensory cues are somewhat discordant they can result in illusory percepts such as the "ventriloquism effect." These biases in stimulus localization are generally accompanied by the perceptual unification of the two stimuli. In the current study, we sought to further elucidate the relationship between localization biases, perceptual unification and measures of a participant's uncertainty in target localization (i.e., variability). Participants performed an auditory localization task in which they were also asked to report on whether they perceived the auditory and visual stimuli to be perceptually unified. The auditory and visual stimuli were delivered at a variety of spatial (0 degrees, 5 degrees, 10 degrees, 15 degrees ) and temporal (200, 500, 800 ms) disparities. Localization bias and reports of perceptual unity occurred even with substantial spatial (i.e., 15 degrees ) and temporal (i.e., 800 ms) disparities. Trial-by-trial comparison of these measures revealed a striking correlation: regardless of their disparity, whenever the auditory and visual stimuli were perceived as unified, they were localized at or very near the light. In contrast, when the stimuli were perceived as not unified, auditory localization was often biased away from the visual stimulus. Furthermore, localization variability was significantly less when the stimuli were perceived as unified. Intriguingly, on non-unity trials such variability increased with decreasing disparity. Together, these results suggest strong and potentially mechanistic links between the multiple facets of multisensory integration that contribute to our perceptual Gestalt. PMID- 15112120 TI - Single joint perturbation during gait: neuronal control of movement trajectory. AB - The aim of this study was to investigate the effect of single joint displacement on the pattern of leg muscle electromyographic (EMG) activity during locomotion. For the first time, unilateral rotational hip or knee joint displacements were applied by a driven orthotic device at three phases of swing during locomotion on a treadmill. The response pattern of bilateral leg muscle activation with respect to the timing and selection of muscles was almost identical for displacements of upper (hip joint) or lower (knee joint) leg. The leg muscle EMG responses were much stronger when the displacement was directed against the physiological movement trajectory, compared with when the displacement was reinforcing, especially during mid swing. It is suggested that these response patterns are designed to restore physiological movement trajectory rather than to correct a single joint position. Displacements released at initial or terminal swing, assisting or resisting the physiological movement trajectory, were followed by similar and rather unspecific response patterns. This was interpreted as being directed to stabilise body equilibrium. PMID- 15112121 TI - Centripetal bias on preparation for smooth pursuit eye movements based on the anticipation. AB - It has been reported that a brief perturbation of a stationary target during fixation induces larger eye movement when monkeys anticipate future smooth pursuit than when they do not. Here, we recorded eye movements in human subjects after briefly perturbing a target and the eccentricity of its initial position was changed under three conditions: (1) subjects anticipated saccades for a target that appeared before; (2) they anticipated smooth pursuit for a target that appeared before; and (3) they anticipated smooth pursuit but did not know beforehand where the target started from. We found that in condition 2 substantial eye movements were induced by the perturbation started moving toward the center. However, weak responses were observed in conditions 1 and 3. These results indicate that ocular responses to brief perturbations of the target at eccentric positions are increased with centripetal bias when human subjects prepare for future smooth pursuit. PMID- 15112130 TI - [Chronic pelvic pain in women]. AB - In this review we discuss the pathophysiological mechanisms and diagnostic criteria of chronic pelvic pain in women. The psychosomatic correlations and important somatic reasons of this complicated disease picture is discussed. For a large number of the patients the diagnostic criteria of somatoform pain disorder are fitting. The therapeutic approaches, which to a large extend are based on experience, are described. Psychosomatic knowledge is very helpful in the management of chronic pelvic pain in women and contributes to a satisfying doctor patient-relationship. PMID- 15112131 TI - [Effects of ovarian stimulation on serum concentrations of lipids and ketone bodies in patients undergoing IVF/ICSI treatment]. AB - OBJECTIVE: The ovarian stimulation increase the estradiol concentration in comparison to the one observed in the menstrual cycle. The aim of the study was to determine the influence of ovarian stimulation on serum concentrations of lipids and ketone bodies in patients undergoing IVF/ICSI treatment. PATIENTS AND METHODS: 50 patients were included in this study. All patients underwent an ovarian stimulation either with recombinant follicle stimulating Hormone (Gonal F, n = 28) or human menopausal gonadotropin (Menogon, n = 22) after pituitary down-regulation with Goserelin or Triptorelin. Five blood samples were taken: before treatment, at the beginning of ovarian stimulation, on the day of HCG injection for the ovulation induction, on the day of follicle aspiration and 14 days after embryo transfer. Afterwards, the samples were analysed with regard to the concentrations of triglyceride, total-cholesterol, HDL-/LDL-cholesterol, acetoacetate, beta-hydroxybutyrate and alpha-ketoglutarate. RESULTS: During the ovarian stimulation, total-cholesterol andA'LDL-cholesterol decreased significantly (p(Chol) < or = 0.001; p(LDL) < or = 0.001), whereas the concentration of HDL-cholesterol increased significantly (p(HDL) < or = 0.001). Furthermore, a significant increase of beta-hydroxybutyrate and alpha ketoglutarate has been detected (p(HBS) = 0.039; p (KG) = 0.023). CONCLUSIONS: The observed changes in serum concentrations of lipids and ketone bodies began with the application of gonadotrophins for stimulation and persisted until the end of the treatment. Therefore, the increase of estradiol, stimulated by gonadotrophins, seems to be responsible for the changes in the serum concentrations of lipids and ketone bodies. All women undergoing ovarian stimulation should be monitored for long-term thrombogenic and atherogenic risks. PMID- 15112132 TI - [Mastitis puerperalis - causes and therapy]. AB - Between 1995 and 2003 a total of 1 827 women suffering from puerperal mastitis was studied. Etiology of the disease was defined, white cell counts and microbiologic cultures were obtained. According to the results the illness was classified and treated. Since in all cases we found either simple congestion or non-infectious inflammation of the breast treatment consisted of intensified emptying of the breast by breast feeding every 2 hours and in some cases additional manual expression of milk. Antibiotics and bromocriptine were not used. In 1 826 cases fever and flue symptoms had resolved within 25-36 hours. Redness and soreness had resolved within 2-3 days. Relapse did not occur and none of the women developed an abscess. One woman had to be treated with antibiotics because the baby refused to nurse at the diseased breast and emptying had to be done by hand. PMID- 15112133 TI - [Breast cancer among Turkish women in Germany - epidemiology and research agenda]. AB - Breast cancer is the leading cause of cancer death and morbidity among women worldwide. In Germany an estimated 46,000 women are diagnosed with breast cancer every year, and 18,000 die of the disease. So far, limited information is available on possible epidemiological and clinical differences between women of German origin and other ethnic or national groups in Germany. Such differences between migrants and the general population could hint to underlying causes of disease and to differential access to medical care. In this paper we present epidemiological information on breast cancer among Turkish women in Germany and describe a future research agenda addressing these questions. PMID- 15112134 TI - [Pain and ethnicity - results of a survey at three internal/gynecological first aid stations in Berlin]. AB - QUESTION: How high is the pain pressure in everyday life of German vs. not German patients, with which troubles do they come to the emergency room, how intense is their pain, how long is it lasting, how many pain areas are named? GROUP OF PATIENTS/METHODS: The study "Utilization of clinical emergency rooms by German patients and migrants" was carried out at first-aid stations of three Berlin hospitals in municipal districts with a high share of foreigners in the population in the form of standardized interviews on following topics: roads to have access to the emergency room, perception and interpretation of pain/troubles, expectations from the first-aid station, further medical care, chronical diseases, self-help measures, social data, migration aspects. RESULTS: An everyday life pressure because of headache and rheumatic pains was named significant more often by migrants of Turkish extraction than by the German women asked. In the age-group 50-65 years the migrants chose significant higher scale values to characterize their pain intensity. Migrants named all in all clearly more pain areas than German patients. The share of patients with pain persisting more than three days was 36.1 % in German women, 45.5 % in migrants of Turkish origin, 45.8 % in women of other ethnicity. CONCLUSIONS: Pain interpretation as well as pain expression are socio-culturally conditioned. Differences between migrants and native patients should lead to take more into consideration the concrete life situation and specific strain in the patients' treatment. PMID- 15112135 TI - [Stereotactic vacuum-assisted breast biopsy - analysis of 166 cases]. AB - OBJECTIVE: Screening mammography (as planned in Germany) will lead to an increasing number of breast biopsies. The purpose of this study was to determine the promise of directional large core biopsy as a patient-protecting therapeutic method. MATERIAL AND METHODS: 166 vacuum assisted, X-ray-guided biopsy procedures were analysed. RESULTS: Histopathologic examination resulted in 75.8 % benign lesions. Atypical proliferation and noninvasive neoplasia was found in 18.6 %, invasive carcinoma in 5.4 % of the biopsies. Complications were few. Neither skin or chestwall injuries, nor pain or intraoperative bleeding caused an abortion. Postoperative we found four cases of bleeding, further on in 28.3 % a superficial, in 3 % a larger and deep hematoma, but in total without any operative revision. No infection was diagnosed. In the average 17.2 (8-31) specimens were removed. After excision of 18 probes the definitive histopathologic diagnosis was clear in all cases, also, the microcalcifications were found. The underestimation rate amounted to 3 of 35 cases. CONCLUSIONS: This clinical study proves stereotactic vacuum-assisted biopsy as a relieable method for analysing indeterminate mammographically detected breast lesions, which shows lower rates of complications than conventional surgical procedures. PMID- 15112136 TI - [Fatal amnioinfusion with previous choriocarcinoma in a parturient woman]. AB - The case of 36-year-old tercipare is described who developed choriocharcinoma in a previous pregnancy. During the first term labour the patient developed cardiac arrest, so reanimation and sectio cesarea was performed. A male new-born was delivered in good condition, but even after intensive therapy and reanimation occurred death of parturient woman with picture of disseminate intravascular coagulopathia (DIK). On autopsy and on histology there was no sign of malignant disease, so it was not possible to connect previous choricarcinoma with amniotic fluid embolism. Maybe was place of choriocarcinoma "locus minoris resistentiae" which later resulted with failure in placentation what was hard to prove. On autopsy we found embolia of lung with a microthrombosis of terminal circulation with punctiformis bleeding in mucous, what stands for DIK. PMID- 15112137 TI - [Neonatal-emergencies: basics in cardiopulmonary resuscitation]. AB - The international guidelines for neonatal resuscitation were recently updated by the American Academy of Pediatrics (AAP), the American Heart Association (AHA) and the International Liaison Committee on Resuscitation (ILCOR). The most important steps in resuscitation of the newly born infant are oxygenation and ventilation, including endotracheal intubation. These fundamental techniques will be emphasized and discussed in a problem-oriented approach. The clinical assessment of the newly born infant is based on a triad of respiration, heart rate and color. If indicated, resuscitation has to be initiated approximately 30 s after birth, i. e. prior to determination of the 1 min. Apgar score and umbilical artery pH. The key to successful neonatal resuscitation is establishment of adequate ventilation; it should commence - after oropharyngeal suctioning and ineffective tactile stimulation - when the heart rate drops < 100 bpm. Clinical evidence supporting the hypothesis that ventilation with room air versus 50 or 100 % oxygen is preferable in terms of neurological outcome is still preliminary and requires further investigation. Chest compressions should be administered if the heart rate remains < 60 bpm (or heart rate 60 to 80 bpm and not rising) despite adequate assisted ventilation. There should be a 3 : 1 ratio of compressions to ventilations to achieve approximately 120 events per minute. Moreover, the international guidelines recommend crystalloid volume expanders (normal saline or Ringer's lactate), red blood cells, sodium bicarbonate and naloxone for cardiopulmonary resuscitation of the newly born infant. PMID- 15112138 TI - [Prenatal diagnosis and outcome of fetal bradyarrhythmias]. AB - BACKGROUND: The purpose of this study was to evaluate the clinical outcome of fetuses with bradyarrhythmias or complete heart block (CHB) in the setting of fetal structural heart disease (CHD) or of maternal collagenosis with and without treatment. METHODS: A retrospective analysis of echocardiographic studies performed in 14 fetuses (mean gestational age 25.5, range 19 - 36 weeks) referred for exclusion or diagnosis of bradyarrhytmias was performed. Maternal SS-A/Ro and SS-B/La antibodies were measured by ELISA. RESULTS: 14 fetuses showed bradyarrhythmias or complete heart block in combination with severe cardiac malformations (n = 7) or with positive maternal antibodies (n = 7). Only one of the fetuses with CHD survived infancy as opposed to 5/7 fetuses with complete atrioventricular block in the setting of maternal collagenosis. Maternal treatment with corticosteroids did not seem to influence the rhythm disorder. CONCLUSION: Fetal echography is a safe method to detect bradyarrhythmias or complete atrioventricular block. When associated with structural heart defects, fetal prognosis is poorer than in combination with maternal collagenosis. PMID- 15112139 TI - [Aseptic meningitis after intrathecal infusion of a parenteral nutrition solution: examples of rare malpositions of central venous catheters in preterm neonates]. AB - BACKGROUND: Central venous catheters are an indispensable part of intensive care. In the management of preterm neonates, these are introduced after puncture of a peripheral subcutaneous vein. The catheters are supposed to be pushed forward with the venous flow. A chest X-ray is usually performed for verification of the position of the catheter. Localization of the tip of the catheter is often very difficult because of its extremely small diameter. METHODS: A malposition of a central venous catheter resulting in intrathecal infusion of parenteral nutrition and subsequent aseptic meningitis in a preterm neonate is presented. Further cases of catheter malpositions leading to arthrography, arteriography and incorrect position in various parts of the venous system are discussed. DISCUSSION AND CONCLUSION: The case reports demonstrate that wire-guided central venous catheters are not only advanced by the forward blood flow but can also inadvertently end up in undesired sites and may thus induce serious side effects. PMID- 15112140 TI - [Fetal monitoring intra partum with cardiotocography and fetal blood sampling analysis and its possible influence on the C-section rate]. AB - In times of rising c-section rates worldwide the obstetricians feel a big responsibility in avoiding a secondary c-section for non-reassuring fetal heart rate. No other experimental method has been established beside the gold standard (cardiotocography and fetal blood sampling) in daily routine until today. The two cases presented demonstrate the fallibility of the "gold standard" in predicting fetal asphyxia and why methodology with better positive predictive value is needed. PMID- 15112141 TI - Neonatal encephalopathy after antidepressant exposure during pregnancy. Herbst F, Gortner L. Paroxetine withdrawal syndrome as differential diagnosis of acute neonatal encephalophathy? Z Geburtsh Neonatol 2003; 207: 232-234. PMID- 15112144 TI - Cancer in older persons: an international issue in an aging world. AB - Persons age 65 years and older bear the greater burden of cancer in the United States and other industrial nations. A cross-national perspective using data from several population-based resources (eg, the NCI Surveillance, Epidemiology, and End Results Program; US Bureau of Census; World Health Organization; and International Association for Research on Cancer) illustrates current and future demographic transitions in America in comparison with six industrial nations, and profiles cancer mortality in older persons across the selected nations--Denmark, France, Italy, Japan, Sweden, and United Kingdom. Mortality rates, age standardized to the world population, are presented for major tumors. US aging and cancer profiles are highlighted. Demographic projections portend a substantial increase in numbers of older persons, and thus, imply resultant increases in cancer incidence and mortality in the elderly. By 2030, there will be larger proportions of persons in the age group most vulnerable to cancer. Information is needed on how age-related health problems affect cancer prevention, detection, prognosis, and treatment. A knowledge base as guidance in management of cancer in the elderly is lacking. Planning for effective prevention measures and improvement of treatment for the elderly is imperative to meet current and future quality cancer care needs. PMID- 15112145 TI - Aging biology and cancer. AB - Epidemiologic analyses of current registries indicate that the majority of patients in the United States with cancer are 65 years old or older. Basic research in both gerontology and oncology has led to an understanding that these processes (normal aging and the development of cancer) have much in common. In this review we attempt to frame specific aspects of cancer biology in the context of normal aging. PMID- 15112146 TI - Geriatric assessment and comorbidity. AB - Elderly persons, a rapidly growing population segment, have an increased incidence of cancer. The older cancer patient's clinical evaluation and treatment is influenced by conditions such as disabilities, comorbidity, and functional status, along with tumor type and stage. These conditions and other geriatric syndromes can be identified by comprehensive geriatric assessment to guide therapy and affect prognosis and quality of life. Comprehensive geriatric assessment involves the medical, functional, affective, social, spiritual, and environmental assessments. The medical assessment, which includes a nutrition, vision, hearing, continence, gait and balance, and cognition evaluation, can provide additional information to performance status and comorbidity. Although there are many assessment domains using several instruments, comprehensive geriatric assessment can be focused and efficient, especially with a multidisciplinary team of nurses, social workers, pharmacists, and other personnel. Comorbid illnesses may have complex interactions, with the underlying cancer influencing cancer diagnosis, disease course, treatment-related side effects, and mortality. Many instruments are available for comorbidity measurement, and retrospective studies in elderly cancer cohorts have shown comorbidity to influence survival. However, the ultimate aim would be to use comorbidity and comprehensive geriatric assessments prospectively in the older cancer patient to help predict the suitability and success of treatment with various antineoplastic modalities. PMID- 15112147 TI - Chemotherapy in the elderly. AB - Persons over the age of 65 years are the fastest growing segment of the US population. In the next 30 years it will comprise over 20% of the population. More than 50% of all cancers occur in this age group and therefore, the total cancer burden is expected to rise. Data are becoming available that will better guide the use of chemotherapy in the older patient population. Understanding the physiologic and functional changes that occur with aging will assist in developing useful strategies in the elderly. Pharmacokinetic data on a broad spectrum of chemotherapeutic agents will be presented. Many of these agents seem to have a beneficial therapeutic index, particularly in older patients. There has also been an increasing trend toward the use of oral chemotherapy. Factors that must be considered in choosing chemotherapy include limitations of saturability of absorption, patient compliance, and the pharmacokinetic and pharmacodynamic changes that occur in older patients. Interpatient variability and age-related changes in drug metabolism are discussed. Careful attention to the physiologic changes with age and dose adjustments necessary for end organ dysfunction (renal, hepatic) are needed to ensure the safe administration of chemotherapy to older patients. PMID- 15112148 TI - Surgery in older patients. AB - Surgery for solid tumors is often withheld from the very elderly because of perceptions that they could not tolerate it physically and often because their life expectancy is considered too short. Data have shown that both of these factors should be carefully considered for the individual patient, since the mortality from most operations, even such major surgeries as liver resections, is no different for the fit elderly than for younger patients. One of the main problems is that the elderly are often not diagnosed and treated early enough to prevent emergency operations that carry a much higher mortality. Many new surgical techniques have made cancer surgery less invasive; this is especially true for stage I and II breast cancer. These advances have increased the potential for curative cancer surgeries that can be offered to all patients regardless of advanced age. PMID- 15112149 TI - The relationship of patient age to the pathobiology of the clonal myeloid diseases. AB - The incidence of the major clonal myeloid diseases, clonal cytopenias, acute, subacute (oligoblastic), and chronic myelogenous leukemia, polycythemia vera, thrombocythemia, and idiopathic myelofibrosis increases in a log-linear manner from young adulthood through advanced age. In older patients, diseases requiring cytotoxic treatment are more difficult and less successful to manage because comorbid conditions and poor performance status are more prevalent, decreasing the tolerance to therapy and increasing the frequency of side effects. This age effect is highlighted by the dramatically less favorable outcome in older than younger patients with acute myeloid leukemia with similar "favorable" cytogenetic changes. In addition, in acute and subacute myeloid leukemia in older patients, the disease is intrinsically more resistant to therapy. Overexpression of drug resistance genes and unfavorable genetic mutations are more prevalent in older patients and provide evidence that acute myeloid leukemia is often qualitatively different in these patients. The gradient of age effects is continuous; the frequency of poor outcome increasing by decade (or less). The decline in survival becomes especially steep as quinquagenarians (50-year-olds) age to nonagenarians (90-year-olds). Although improved drug schedules have led to significant improvements in event-free survival in younger patients, these improvements have been far less evident in older patients. New approaches, especially the development of drugs aimed at new targets, will be required to obtain a high frequency of long-term remissions in older patients. Agents that reverse inherent cellular drug resistance, farnesyltransferase inhibitors, BCL-2 inhibitors, and FLT3 inhibitors are early examples of such approaches. PMID- 15112150 TI - Lymphoma and myeloma in older patients. AB - Lymphoma and myeloma represent an increasingly important cause of morbidity and mortality in the older patient group. Studies to date, while limited, suggest that there may be differences in the underlying biology of the tumor cell of some lymphoid malignancies in younger versus older patients. Significant new data will be required to document and elucidate the differences and use them to tailor therapy to discrete disease entities. Increasing numbers of clinical trials are addressing the issues of treatment in the elderly, particularly in the aggressive non-Hodgkin's lymphomas. However, much work remains in improving the participation of older patient groups in current clinical trials and in overcoming inherent treatment biases related to concerns of toxicity simply as a function of age, both of which limit application of existing successful therapy to older patient groups. PMID- 15112151 TI - Gastrointestinal cancer in older patients. AB - More than two thirds of gastrointestinal cancers occur in persons 65 years of age or older. The symptoms and presentation in these older individuals appear similar to those of persons of younger age. Although treatments for these cancers have been developed primarily in younger patients, greater expertise over time has permitted similarly safe and efficacious therapy to be extended to older age groups. The majority of gastrointestinal cancers are located in the colon and rectum. Preventative strategies for colorectal cancer are quickly evolving, with the beneficial effect of long-term use of aspirin and estrogen having their greatest impact in the elderly population. The increased acceptance of colonoscopy for screening patients for colorectal cancer will be of greatest benefit in older individuals, who have a higher incidence of proximal neoplasms than younger individuals. Adjuvant therapy for both colon and rectal cancer is underutilized in elderly patients, despite such life-saving treatments resulting in similar survival prolongation, as well as toxicity profiles, as in their younger counterparts. There is a paucity of information concerning the treatment of elderly patients with other gastrointestinal malignancies. PMID- 15112152 TI - Aerorespiratory tract cancer in older patients. AB - Lung cancer and head and neck cancer present distinctive management challenges in the elderly population. Cumulative tobacco exposure is a risk factor for both types of cancer. Smoking-related comorbidities, including cardiovascular disease and chronic obstructive pulmonary disease, will also increase with cumulative tobacco exposure and can complicate surgical or radiotherapeutic management. Some differences in natural history, such as a greater tendency toward localized disease in non-small cell lung cancer and a lesser correlation with tobacco/alcohol exposure in head and neck cancer, have been noted for older patients. However, standard treatment modalities do appear to maintain efficacy in the elderly population. If comorbidities and physiologic age (rather than strict chronologic age) are taken into account, effective interventions can be devised with benefits similar to those seen in younger populations. Although prevention remains the mainstay of management of aero-respiratory cancers, carefully planned treatment can result in therapeutic benefit with maintenance of quality of life. PMID- 15112153 TI - Breast cancer in older patients. AB - Breast cancer is a common problem and a major health concern in our growing geriatric population. Older breast cancer patients are at risk for less than standard management, the appropriateness of which is difficult to discern. Breast tumors tend to have less aggressive characteristics. In addition, planning therapy is not always straightforward because older patients may present with comorbid illnesses and frailty that limit therapeutic choices. Standard management approaches should always be considered first. Here, we outline some data supporting standard treatment for breast cancer in older women. We also describe other options that can be considered in circumstances when the standard treatment is not possible. For instance, primary treatment with tamoxifen or an aromatase inhibitor is justifiable in a patient who is unfit for surgery and axillary dissection may be unnecessary in a patient who is obviously unfit for adjuvant chemotherapy. Adjuvant therapies should be considered, weighing risks and benefits for each patient, though the threshold for using chemotherapy may be higher. The goals in treating metastatic breast cancer in an older patient are not different than for younger patients. PMID- 15112154 TI - Genitourinary cancer in the elderly. AB - Genitourinary malignancy comprises nearly half of the cancers diagnosed in men, and the incidence of this group of cancers increases with age. The key to successful management is to define appropriate goals (cure v palliation) based on the natural history and extent of disease, physiology and life expectancy of the patients, and cost-benefit ratio of treatment options. Of particular importance, the chance for cure should not be sacrificed because of age-based considerations in early-stage cancers of the prostate or kidney nor in the early stage of locally advanced urothelial malignancy. PMID- 15112155 TI - Multiple primary malignancies. AB - As the incidence and prevalence of cancer increase with age, it is reasonable to expect that the incidence and prevalence of multiple primary malignancies will increase as well. We ask here whether age is a risk factor for multiple primary malignancies and whether a special phenotype of older individual at increased risk of cancer may be identified. The main sources of information are autopsy series and tumor registries, which indicate that the prevalence and risk of multiple primary malignancies increase with age as expected, but that it is not possible to identify a particular phenotype that is at increased risk of multiple primary malignancies, among older individuals. It is also noted that the risk of endometrial cancer in association with breast cancer seems to increase in women aged 70 and older. Our conclusions are affected by the limitation of the sources themselves. These include for autopsy studies referral bias and overestimate of risk due to occult neoplasms, and for tumor registries, the quality and comprehensiveness of the registry and the evolution of diagnostic techniques overtime. Prospective follow-up of older cancer patients, involving measurement of circulating cytokines, whose concentration increases with age, and study of the genotype of older individuals, may provide new information about whether certain older individuals are at increased risk for multiple primary neoplasms. PMID- 15112156 TI - End of life issues in older patients. AB - Elderly patients with cancer at the end of their life have very unique features associated with their care. As the population of the United States ages, these issues will involve more patients, and end-of-life care has the possibility of consuming more health care costs. With regard to unique care issues, the elderly with cancer at the end of their life are more at risk for untreated pain and depression and are more likely to commit suicide than their younger counter parts. Advance directives are important to discuss with patients and their families so that the patient can designate an individual as the health care proxy and so that discussion around end-of-life care can occur before patients clinically decline so that patients' wishes are known and futile care can be avoided. Hospice care is a Medicare-sponsored program that elderly and nonelderly patients can enter for the last 6 months of their lives. This article discusses barriers to hospice referral. PMID- 15112157 TI - Elder law, Medicare, and legal issues in older patients. AB - Elder law is a specialty area of law directed at the specific needs of older clients and clients with disabilities. This review discusses major legal issues faced by older patients, their families, and their health care providers, including the capacity to consent to treatment, advance directives, and surrogate decision-making. In addition, public benefit programs such as Medicare, Social Security Disability Insurance, Medicaid, and Supplemental Security Income are examined. Available options for long-term care, both institutional and community based are also highlighted. Finally, elder abuse is discussed. All who provide care to an elder must be aware of the potential signs of abuse and the remedies available. PMID- 15112166 TI - Restless legs syndrome in patients on dialysis. AB - The past decade has seen an explosion of interest in both idiopathic and secondary restless legs syndrome (RLS). Secondary RLS occurs in patients with uremia, pregnancy, and iron deficiency. Patients experience an irresistible urge to move the legs that is worse during inactivity and at night. RLS affects 6.6% to 62% of patients on long-term dialysis therapy and is associated with a greater mortality risk. The wide range of reported prevalence is explained in part by variations in methods of diagnosis. The International Restless Legs Syndrome Study Group defined diagnostic criteria that have improved the quality of RLS research. Advanced neurological imaging techniques suggest the pathophysiological state of idiopathic RLS involves dysfunction of subcortical areas of the brain. Dopaminergic pathways and neuronal iron handling have been implicated. Limited studies of patients with uremic RLS suggested similar mechanisms, but anemia, hyperphosphatemia, and psychological factors also may have a role. The few clinical trials in uremic RLS suggest that treatment should involve the reduction of potential exacerbating agents (tricyclic antidepressants, selective serotonin uptake inhibitors, lithium, and dopamine antagonists), correction of anemia (with erythropoietin and iron), and use of levodopa or dopamine agonists. Other agents shown to be of benefit in idiopathic RLS can be tried, but may be limited by side effects in patients with uremia (benzodiazepines, opioids, gabapentin, carbamazepine, and clonidine). Symptoms of uremic RLS will disappear within a few weeks of successful renal transplantation. The progress made to date in unraveling the pathophysiological state of uremic RLS should stimulate additional research toward targeted therapy. PMID- 15112167 TI - Strategies for preservation of ovarian and testicular function after immunosuppression. AB - Gonadal toxicity as a side effect of cyclophosphamide therapy is a common long term problem in the treatment of a variety of glomerular diseases. In both men and women treated with cyclophosphamide, the consequences of infertility can have great physical and emotional consequences; thus, this issue often has a critical role in the decision to decline treatment with cyclophosphamide. There exists a critical need for strategies for preservation of fertility in both men and women who require treatment with cyclophosphamide. This review explores emerging therapeutic options in this arena, which include sperm and oocyte cryopreservation, medical treatments such as testosterone therapy for men and gonadotropin-releasing hormone agonist therapy for both men and women, and, finally, the relatively new strategy of germ-cell transplantation for both ovarian and testicular tissue, which still remains in the experimental stages. PMID- 15112168 TI - Achievement of target blood pressure levels in chronic kidney disease: a salty question? AB - A large body of evidence supports the validity of lowering blood pressure (BP) to prevent cardiovascular (CV) disease in the general population. This issue becomes even more critical in renal patients because they carry a greater CV risk across the entire spectrum of chronic kidney disease (CKD). In these patients, achievement of lower BP levels also is fundamental to limit the progression of renal damage, especially in the presence of significant proteinuria. Although expert panels have strongly recommended to intensively decrease BP in patients with CKD, management of hypertension in these patients remains inadequate. Armed with the knowledge of the extreme salt-sensitivity of BP in patients with CKD, it is reasonable to hypothesize that more aggressive treatment of volume expansion can be helpful. Nevertheless, although abundant literature has evidenced that dietary sodium restriction decreases BP levels in patients with essential hypertension, no large and prospective study has been conducted to date on this issue in patients with CKD. A potential reason is the low compliance of patients with CKD to dietary prescriptions; however, this problem can be overcome by specific counseling. Alternatively, loop diuretics administered at a high dose should represent the cornerstone of therapy, but, again, well-designed studies verifying the effectiveness of these agents in a large CKD population are still awaited. Nephrologists seem to be reluctant to adequately administer diuretics because of the fear of adverse events. Conversely, the major detrimental effect, that is, excessive hypovolemia, can be prevented if daily body weight loss is limited to 0.3 to 0.5 kg during the initial period of treatment. PMID- 15112169 TI - Heritability of GFR and albuminuria in Caucasians with type 2 diabetes mellitus. AB - BACKGROUND: Elevated urinary albumin excretion and decreased glomerular filtration rate (GFR) are risk factors for cardiovascular death and end-stage renal disease in individuals with type 2 diabetes mellitus (DM). METHODS: To determine the extent of familial aggregation of GFR and urine albumin-creatinine ratio (ACR), we calculated heritability (h2) estimates by using a variance component approach. RESULTS: Among 662 participants with DM from 310 families (422 DM-concordant sibling pairs), 51.8% (n = 343) were women, mean age was 62.3 +/- 9.2 (SD) years (median, 62.6 years), diabetes duration was 10.8 +/- 7.6 years (median, 9 years), GFR was 67.6 +/- 19.0 mL/min (median, 64.7 mL/min), and urine ACR was 139.7 +/- 631.4 mg/g (median, 13.1 mg/g). Estimated h2 of GFR was 0.75 +/ 0.10 (P < 0.0001) after adjusting for age, sex, mean arterial blood pressure, medications, and hemoglobin A(1c) level. These covariates accounted for only 2% of the total phenotypic variation in log GFR. Similarly, estimated h2 of ACR was 0.46 +/- 0.12 (P < 0.0001) when adjusting for these covariates, with covariates contributing only 9% of phenotypic variation. CONCLUSION: These data provide strong evidence that among Caucasians with type 2 diabetes, GFR and urine ACR show strong familiality, suggesting that genetic factors exhibit significant influences. Given their biological and clinical importance and the similarity of these estimates with other cardiovascular disease- and DM-related traits, efforts to map genes that influence GFR and urine ACR levels should have increased importance. PMID- 15112170 TI - Effect of N-acetylcysteine for prevention of contrast nephropathy in patients with moderate to severe renal insufficiency: a randomized trial. AB - BACKGROUND: The effect of N-acetylcysteine (NAC) to prevent contrast nephropathy (CN) in patients with moderate to severe renal insufficiency undergoing coronary angiography or interventions is not clear. METHODS: This is a prospective, open label, randomized, controlled trial. Ninety-one consecutive patients with a serum creatinine level of 1.69 to 4.52 mg/dL (149 to 400 micromol/L) undergoing coronary procedures were recruited and randomly assigned to administration of either oral NAC, 400 mg, thrice daily the day before and day of the contrast procedure (the NAC group) or no drug (the control group). Serum creatinine was measured before and 48 hours after contrast exposure. The primary end point of this study was the development of CN, defined as an increase in serum creatinine concentration of 0.5 mg/dL or greater (> or =44 micromol/L) or a reduction in estimated glomerular filtration rate (GFR) of 25% or greater of the baseline value 48 hours after the procedure. RESULTS: There were no significant differences between the 2 groups (46 patients, NAC group; 45 patients, control group) in baseline characteristics or mean volume of contrast agent administered. Six patients (13.3%) in the control group and 8 patients (17.4%) in the NAC group developed CN (P = 0.8). Serum creatinine levels increased from 2.27 +/- 0.54 to 2.45 +/- 0.65 mg/dL (201 +/- 48 to 217 +/- 57 micromol/L; P = 0.003) in the NAC group and 2.37 +/- 0.61 to 2.40 +/- 0.70 mg/dL (210 +/- 54 to 212 +/- 62 micromol/L; P = 0.6) in the control group. The increase in serum creatinine levels between the 2 groups had no difference (P = 0.7). Estimated GFR decreased from 30.3 +/- 8.4 to 28.1 +/- 8.4 mL/min (P = 0.01) in the NAC group and 28.4 +/- 8.6 to 27.5 +/- 8.8 mL/min (P = 0.3) in the control group. The decline in estimated GFR between the 2 groups had no difference (P = 0.7). CONCLUSION: In the current study, oral NAC had no effect on the prevention of CN in patents with moderate to severe renal insufficiency undergoing coronary angiography or interventions. However, the sample size of our present study is small. Our findings highlight the need for a large-scale, randomized, controlled trial to determine the exact beneficial effect of NAC. PMID- 15112171 TI - Urinary excretion of interleukin-6 correlates with proteinuria in acute Puumala hantavirus-induced nephritis. AB - BACKGROUND: Nephropathia epidemica (NE) is a mild type of hemorrhagic fever with renal syndrome caused by Puumala Hantavirus. Cytokines are thought to have an important role in the pathogenesis of NE. The aim of this study is to evaluate whether cytokines contribute to renal involvement in NE. METHODS: Overnight urinary excretion of interleukin-1beta (IL-1beta), IL-1 receptor antagonist (IL 1Ra), IL-6, tumor necrosis factor-alpha (TNF-alpha), albumin, immunoglobulin G (IgG), and alpha1-microglobulin and quantitative 24-hour urinary protein excretion were measured for 3 consecutive days from 70 hospitalized patients with acute NE (49 men, 21 women; age, 15 to 70 years; median age, 39 years). Plasma levels of the respective cytokines also were measured. Urinary collections were repeated after 1 year. The control group for blood samples included 400 healthy blood donors. RESULTS: Maximum median urinary IL-6 excretion in the acute phase of NE was increased compared with values detected after 1 year (49.5 versus 0.7 pg/min; P < 0.001). Correspondingly, maximum median plasma IL-6 concentration in patients was increased compared with controls (14.6 versus 1.2 pg/mL; P < 0.001). Urinary IL-6 excretion correlated with urinary albumin, IgG, and protein excretion (r = 0.79; P < 0.001; r = 0.76; P < 0.001; and r = 0.65; P < 0.001, respectively), but not plasma IL-6 levels (r = 0.18; P = 0.148). CONCLUSION: Plasma IL-6 concentrations and urinary IL-6 excretion were markedly increased in patients with acute NE, but there was no correlation between plasma and urinary IL-6 levels. The high urinary IL-6 levels might reflect local production of this proinflammatory cytokine in the kidneys during acute infection. PMID- 15112172 TI - Clinical characteristics of patients developing ARF due to sepsis/systemic inflammatory response syndrome: results of a prospective study. AB - BACKGROUND: Acute renal failure (ARF) in patients with sepsis provokes high mortality and financial cost. In this prospective study, we collected characteristics of patients in the intensive care unit (ICU) who developed sepsis/systemic inflammatory response syndrome (SIRS) to analyze differences between those who subsequently did or did not develop ARF. METHODS: All patients admitted to the ICU of the University Hospital Gent, Belgium, between January 1, 2001, and December 31, 2001, who developed sepsis/SIRS were included if they had a serum creatinine level less than 2 mg/dL (<177 micromol/L). RESULTS: Of 2,442 patients admitted to the ICU, 257 patients developed sepsis/SIRS. Of those, 29 patients (11%) developed ARF. In a univariate analysis, age, central venous pressure (CVP), and serum creatinine and blood urea nitrogen levels were greater (P = 0.003, P = 0.006, P < 0.001, and P < 0.001, respectively), whereas mean arterial and diastolic blood pressures, 24-hour urinary output, arterial pH, bicarbonate level, thrombocyte count, albumin level, and prothrombin time were lower (P = 0.05, P = 0.004, P = 0.005, P = 0.03, P = 0.009, P = 0.037, P = 0.05, and P = 0.006, respectively) in the ARF group. Prevalence of diabetes, sex, and need for ventilation were not different between the ARF and no-ARF groups, but in the ARF group, diuretic use, vasopressor use, and presence of primary hepatic failure were more prevalent (P = 0.001 for each). In a multivariate analysis, age, serum creatinine level, CVP, and presence of liver failure significantly contributed to a logistic regression model for ARF. CONCLUSION: Several parameters already were disturbed at the first day of SIRS/sepsis in patients who later developed ARF. Older age, elevated serum creatinine level despite elevated CVP, and presence of hepatic failure are predictive for ARF in septic patients. PMID- 15112173 TI - Unreferred chronic kidney disease: a longitudinal study. AB - BACKGROUND: Chronic kidney disease (CKD) is a major health problem. A better understanding of the epidemiological characteristics of the different stages of CKD and the associated adverse outcomes is needed to establish and implement appropriate management strategies. METHODS: A serum creatinine (SCr) level of 2.03 mg/dL or greater (> or =180 micromol/L) in men and 1.53 mg/dL or greater (> or =135 micromol/L) in women was used to identify patients with moderate to severe CKD in a predominantly Caucasian area of the United Kingdom. Patients who were unknown to renal services were identified and followed up to establish survival, rate of referral, and change in glomerular filtration rate (GFR). RESULTS: The prevalence of CKD defined by SCr cutoff values was 5,554 per million population (pmp). Median calculated GFR of the cohort was 28.5 mL/min/1.73 m2 (range, 4.1 to 42.8 mL/min/1.73 m2), and median age was 83 years (range, 18 to 103 years). A total of 84.8% of patients were unknown to renal services. During a mean follow-up of 31.3 months, 8.1% of patients were referred. Median survival of the unreferred population was 28.1 months. Cardiovascular disease, cancer, and infection were the most common causes of death. Male sex, low GFR, and nonreferral were associated with poor outcome. The majority of unreferred patients had stable renal function. The incidence of new unreferred CKD during the first year of follow-up was 2,435 pmp, such that the prevalence remained stable at 4,910 pmp. Significant anemia (hemoglobin < 11 g/dL [<110 g/L]) was seen in 27.5% of the unreferred cohort. CONCLUSION: Referral of all patients with CKD is unrealistic and inappropriate. Management strategies aimed at improving adverse outcomes need to take account of this and be developed and implemented through collaboration between primary care and secondary care. PMID- 15112174 TI - Association of kidney function and hemoglobin with left ventricular morphology among African Americans: the Atherosclerosis Risk in Communities (ARIC) study. AB - BACKGROUND: Left ventricular (LV) hypertrophy (LVH) is present in more than 70% of patients with kidney failure. Hypertension and anemia, 2 strong risk factors for LVH, are common in patients with moderate kidney dysfunction. It is unknown whether kidney function and lower hemoglobin levels are associated with LV morphological characteristics among African Americans with moderate kidney dysfunction, independent of blood pressure. METHODS: As part of the Atherosclerosis Risk in Communities Study, 1,968 African Americans aged 50 to 75 years underwent an echocardiogram after 6 to 9 years of follow-up. LV mass was indexed (LVMI) to body surface area. Glomerular filtration rate (GFR) was estimated from calibrated serum creatinine levels measured at baseline and after 3 years (<30 mL/min/1.73 m2 excluded). Hemoglobin was measured at baseline and after 3 years. Blood pressure was measured every 3 years. RESULTS: A mean GFR of 30 to 59 mL/min/1.73 m2 was associated with a 15.3 g/m2 greater LVMI, 0.9 mm greater posterior wall thickness, and 1.0 mm greater interventricular septal thickness compared with a GFR of 90 mL/min/1.73 m2 or greater. These associations remained after adjustment for age, sex, hemoglobin level, blood pressure, and other covariates. Wall thickness and LVMI increased with lower GFR less than 75 mL/min/1.73 m2 (5.5 g/m2 greater LVMI [95% confidence interval (CI), 0.9 to 10.1] per 10 mL/min/1.73 m2 lower GFR). A lower hemoglobin level was associated with greater LV diameter (0.5 mm [95% CI, 0.2 to 0.7] per 1 g/dL [10 g/L]), but not wall thickness, after adjustment. CONCLUSION: These results in a general population sample of middle-aged African Americans suggest that moderately decreased kidney function independently predicts greater LV wall thickness, and lower hemoglobin level independently predicts greater LV diameter. These findings may explain, in part, the high risk for cardiovascular disease observed among individuals with kidney disease. PMID- 15112175 TI - Clinical significance of cardiac troponin T levels in chronic kidney disease patients: predictive value for cardiovascular risk. AB - BACKGROUND: Recent studies have shown that cardiac troponin T (cTnT) levels predict cardiovascular mortality in hemodialysis patients. The aim of this study is to analyze the prognostic value of cTnT levels in stable patients with moderate chronic kidney disease (CKD). METHODS: One hundred seventy-six outpatients were included; 128 patients had an estimated glomerular filtration rate (GFR) less than 60 mL/min (<1 mL/s; CKD group), and 48 patients, 60 mL/min or greater (> or =1 mL/s; control group). Medical records for cardiovascular factors were recorded. Analytical parameters and cardiac markers were analyzed. Patients were followed up prospectively, and the end points were fatal and nonfatal cardiovascular events. RESULTS: cTnT levels were undetectable in all control group patients (cTnT < 0.01 ng/mL), and only 20 patients in the CKD group had detectable values (>0.01 ng/mL). Patients with detectable cTnT levels were older (P = 0.005), had a greater prevalence of congestive heart failure (CHF; P = 0.000) and left ventricular hypertrophy (P = 0.001), more anemia (P = 0.008), and a lower GFR (P = 0.008) than the rest of the patients in the CKD group. After a mean follow-up of 12.9 months, 20 patients in the CKD group and 1 patient in the control group experienced a cardiovascular event. Age (odds ratio [OR], 1.11), previous coronary artery disease (OR, 2.49), CHF (OR, 7.36), hemoglobin level (OR, 0.70), GFR (OR, 0.94), pulse pressure (OR, 1.02), and cTnT level (OR, 12.34) predicted cardiovascular events on univariate Cox analysis. CONCLUSION: In patients with CKD, cTnT level is a predictor of cardiovascular events. PMID- 15112176 TI - Endothelial dysfunction and increased carotid intima-media thickness in patients with autosomal dominant polycystic kidney disease. AB - BACKGROUND: Cardiovascular problems are a major cause of morbidity and mortality in patients with autosomal dominant polycystic kidney disease (ADPKD). Endothelial dysfunction (ED) and intima-media thickness (IMT) are predictors for the development and progression of atherosclerosis. In the present study, ED and IMT were investigated in patients with ADPKD. METHODS: Fifteen hypertensive and 16 normotensive patients with ADPKD with preserved renal function, 16 patients with essential hypertension, and 24 healthy subjects were included in the study. Endothelial function of the brachial artery was evaluated by means of high resolution vascular ultrasound. Endothelial-dependent dilatation (EDD) was assessed by establishing reactive hyperemia, and endothelial-independent dilatation was determined by using sublingual isosorbide dinitrate. Carotid IMT was measured by means of high-resolution vascular ultrasound. RESULTS: EDD was significantly worse in hypertensive patients with ADPKD compared with patients with essential hypertension (9.1% +/- 4.1% versus 12.4% +/- 4.6%; P < 0.05) and even in normotensive patients with ADPKD compared with healthy subjects (13.1% +/ 5.2% versus 18.1% +/- 8.1%; P < 0.01). Moreover, carotid IMT was significantly greater in both hypertensive (0.71 +/- 0.10 mm; P < 0.01) and normotensive (0.57 +/- 0.14 mm; P < 0.001) patients with ADPKD compared with healthy subjects (0.45 +/- 0.10 mm). CONCLUSION: Both hypertensive and normotensive patients with ADPKD show significant ED and increased IMT, which are predictors of atherosclerosis. PMID- 15112177 TI - The presence of frailty in elderly persons with chronic renal insufficiency. AB - BACKGROUND: Frailty has been defined as a tool to define individuals who lack functional reserve and are at risk for functional decline. We hypothesized that chronic renal insufficiency (CRI) would be associated with a greater prevalence of frailty and disability in the elderly. METHODS: This cross-sectional analysis used baseline data collected from the Cardiovascular Health Study, which enrolled 5,888 community-dwelling adults aged 65 years or older from 4 clinical centers in the United States. Renal insufficiency is defined as a serum creatinine level of 1.3 mg/dL or greater (> or =115 micromol/L) in women and 1.5 mg/dL or greater (> or =133 micromol/L) in men. Frailty is defined by the presence of 3 of the following abnormalities: unintentional weight loss, self-reported exhaustion, measured weakness, slow walking speed, and low physical activity. Disability is defined as any self-reported difficulty with activities of daily living. RESULTS: Among 5,808 participants with creatinine levels measured at entry, 15.9% of men (n = 394) and 7.6% of women (n = 254) had CRI. Prevalences of frailty (15% versus 6%; P < 0.001) and disability (12% versus 7%; P = 0.001) were greater in participants with CRI compared with those with normal renal function. After multivariate adjustment for comorbidity, CRI remained significantly associated with frailty (odds ratio, 1.76; 95% confidence interval, 1.28 to 2.41), but not disability (odds ratio, 1.26; 95% confidence interval, 0.94 to 1.69). CONCLUSION: Elderly persons with CRI have a high prevalence of frailty, which may signal their risk for progression to adverse health outcomes. If confirmed in other studies, identification of frailty in patients with CRI may warrant special interventions to preserve their independence, quality of life, and survival. PMID- 15112178 TI - Physical and sexual function in women with chronic kidney disease. AB - BACKGROUND: Cross-sectional studies suggest an association between functional status and chronic kidney disease (CKD). Whether physical function deteriorates with progression of CKD is unknown. METHODS: To determine associations among CKD, physical function, and sexual function in women, we conducted cross-sectional and longitudinal analyses of 2,761 women enrolled in the Heart and Estrogen/Progestin Replacement Study. Physical and sexual function were evaluated using the Duke Activity Status Index (DASI) and the Sexual Problems Scale of the Medical Outcomes Study, respectively. Glomerular filtration rate (GFR) was estimated using the Modification of Diet in Renal Disease regression equation. In addition to analyses across the spectrum of GFR, CKD was categorized as mild (estimated GFR, 45 to 60 mL/min/1.73 m2), moderate (estimated GFR, 30 to 44 mL/min/1.73 m2), and severe (estimated GFR, <30 mL/min/1.73 m2) according to a modification of recently established classification guidelines. RESULTS: Mean age of study participants was 67 +/- 7 years, and mean estimated GFR was 61 +/- 14 mL/min/1.73 m2. In unadjusted analyses, mean baseline DASI score was 10 points lower in women with an estimated GFR less than 30 mL/min/1.73 m2 than in women with an estimated GFR of 60 mL/min/1.73 m2 or greater (P < 0.0001). Estimated GFR remained significantly associated with DASI score after multivariable adjustment. In longitudinal analyses, a decline in estimated GFR was associated with a significant decline in DASI score independent of baseline estimated GFR and other factors. There were no significant associations between estimated GFR and psychosocial aspects of sexual function. CONCLUSION: CKD is associated with impaired physical function, and a decline in estimated GFR is associated with a decline in physical function. PMID- 15112179 TI - Doxercalciferol safely suppresses PTH levels in patients with secondary hyperparathyroidism associated with chronic kidney disease stages 3 and 4. AB - BACKGROUND: Calcitriol lowers parathyroid hormone (PTH) levels in patients with chronic kidney disease (CKD) stages 3 and 4, but its use is limited by a low therapeutic index and concerns regarding hypercalcemia and acceleration of kidney disease. We evaluated doxercalciferol (1alpha-hydroxyvitamin D2) as an alternative therapy in a randomized, double-blinded, placebo-controlled, multicenter trial. METHODS: Fifty-five adults with stage 3 or 4 CKD and an intact PTH (iPTH) level greater than 85 pg/mL (ng/L) completed 8 baseline weeks, followed by 24 weeks of oral therapy with doxercalciferol or placebo. Pretreatment demographics and biochemical features did not differ between groups. Dosages were increased gradually if iPTH level was not decreased by 30% or greater and serum calcium and phosphorus levels were stable. Regular monitoring included plasma iPTH, serum calcium and phosphorus, urinary calcium, bone specific serum markers, and serum lalpha,25-dihydroxyvitamin D levels. Glomerular filtration rate (GFR) was measured before and after treatment. RESULTS: Mean plasma iPTH level decreased by 46% from baseline after 24 weeks of doxercalciferol treatment (P <0.001), but was unchanged with placebo. After 6 weeks, iPTH level reductions with doxercalciferol treatment exceeded those with placebo at all subsequent intervals (P <0.001). No clinically significant differences in mean serum calcium or phosphorus or urinary calcium levels or incidence of hypercalcemia, hyperphosphatemia, or hypercalciuria were noted between groups. Serum C- and N-telopeptide and bone-specific alkaline phosphatase levels decreased with doxercalciferol treatment relative to both baseline and placebo (P <0.01). Adverse-event rates and changes in GFR did not differ between groups. CONCLUSION: Doxercalciferol is safe and effective in controlling secondary hyperparathyroidism of patients with CKD stages 3 and 4. PMID- 15112180 TI - The effect of contraindications and patient preference on dialysis modality selection in ESRD patients in The Netherlands. AB - BACKGROUND: Patients with end-stage renal disease (ESRD) who are about to start long-term dialysis therapy are faced with the question of modality choice. The aim of the current study is to determine the influence of different factors on long-term dialysis modality choice. METHODS: As part of a large Dutch prospective multicenter study, the Netherlands Cooperative Study on the Adequacy of Dialysis, we consecutively included all new patients with ESRD. Nephrologists indicated the most important reason for the modality selection. RESULTS: Of 1,347 included patients, 36% (n = 483) had a contraindication to either peritoneal dialysis (PD) or hemodialysis (HD) therapy. Eighty percent (n = 386) of all contraindications were directed to PD therapy. The most frequently mentioned contraindication was a social one; ie, the expected incapability of patients to perform PD exchanges themselves. Patients with contraindications were older, had more comorbidity, and lived alone more often compared with patients without contraindications. In patients without contraindications (64%), modality choice was based on patient preference. Older age increased the odds of choosing HD, whereas receipt of predialysis care was associated with a lower preference for HD. CONCLUSION: Older age was associated with more contraindications to PD therapy and stronger patient preference for HD therapy. An elderly patient therefore was more likely to start with HD therapy. Results from the current study suggest that an increase in provision of predialysis care, in combination with a reduction in social contraindications to PD therapy, may be associated with an increase in likelihood of starting with PD therapy. In a time of an aging population, increasing demand on dialysis capacity, and limited amount of financial supplies, we may reconsider current strategies to provide future patients with ESRD the possibility to start with the dialysis modality they prefer. PMID- 15112181 TI - Restless legs symptoms among incident dialysis patients: association with lower quality of life and shorter survival. AB - BACKGROUND: Symptoms of restless legs are common among patients treated with long term hemodialysis. We investigated the relation between symptoms of restless legs, quality of life, and survival among incident hemodialysis and peritoneal dialysis patients. METHODS: This report includes 894 dialysis patients who responded to an item regarding severity of restless legs symptoms in the Choices for Healthy Outcomes in Caring for End-Stage Renal Disease Study. We examined participant responses both as categorized by the 5-point symptoms of restless legs item and classified with severe symptoms of restless legs if they reported they were very or extremely bothered by symptoms. Independent predictors of restless legs symptoms were estimated using logistic regression. The association of restless legs symptoms with quality of life was examined by means of mixed model regression, and adjusted mortality risk was estimated by means of Cox proportional hazards. RESULTS: Fifteen percent of dialysis patients were classified as having severe restless legs. Those characterized by severe restless legs were significantly more likely to be administered benzodiazepines, antidepressants, and antiepileptics. Age and diabetes mellitus were independently associated with severe symptoms of restless legs. Symptoms of restless legs were associated with lower Physical and Mental Component Scores, vitality, bodily pain, and sleep quality (all P < 0.001). After adjustment, severe symptoms of restless legs were significantly associated with an increased mortality hazard ratio of 1.39 (95% confidence interval, 1.08 to 1.79). CONCLUSION: Symptoms of restless legs were associated with lower quality of life and increased risk for death. The effect of treatment for symptoms of restless legs on quality of life and survival awaits additional study. PMID- 15112182 TI - Efficacy and side effects of intradermal hepatitis B vaccination in CAPD patients: a comparison with the intramuscular vaccination. AB - BACKGROUND: This prospective randomized study aims to assess the effectiveness of intradermal (ID) hepatitis B virus (HBV) vaccination in patients on continuous ambulatory peritoneal dialysis (CAPD) therapy. METHODS: Sixty patients were randomly divided into 2 groups. The ID group was treated with 5 microg of recombinant HBV vaccine intradermally every week for a total of 10 doses, and the intramuscular (IM) group, with 20 microg intramuscularly at 0, 1, and 6 months. RESULTS: ID HBV vaccination was associated with a greater seroconversion rate (81.5% versus 62.1%), although the difference did not reach statistical significance (P = 0.14). The cumulative seroconversion rate was significantly greater with ID vaccination by 6 months after the first vaccine dose (P = 0.03). There was no difference between the 2 groups in time required to convert, peak antibody to HBV surface antigen (anti-HBs), and proportion of patients with anti HBs levels maintained at greater than 10 mIU/mL or 100 mIU/mL in the 2-year observation period. Although the ID group achieved a peak anti-HBs titer significantly earlier than the IM group (P = 0.001), we found a significant trend for the ID group to achieve a lower peak anti-HBs titer (chi-square test for trend, P = 0.005). The incidence of local reactions was significantly greater with ID immunization; however, reactions were mild and transient. CONCLUSION: ID HBV vaccination is associated with significant improvement in seroconversion rate in CAPD patients at 6 months, but this difference diminishes at 2 years. Larger studies are warranted to confirm this finding. PMID- 15112183 TI - Typical MPGN with few urinary abnormalities. AB - We describe the concurrent appearance of membranoproliferative glomerulonephritis (MPGN) in renal biopsy samples and normal urinary sediment without hematuria, proteinuria, or renal dysfunction in 1 child and asymptomatic microscopic hematuria without significant proteinuria or renal dysfunction in 2 children who were subsequently followed up for many years. The only other abnormality detected was hypocomplementemia. This is the first report of biopsy-proven typical MPGN in patients with few urinary abnormalities. A renal biopsy should be considered in children with hypocomplementemia, regardless of urinalysis findings, to exclude MPGN. PMID- 15112184 TI - Treatment of secondary pulmonary hypertension with bosentan and its pharmacokinetic monitoring in ESRD. AB - Pulmonary hypertension (PH) is a rare disease with a very poor prognosis. Certain pharmacologic approaches, which reduce pulmonary arterial pressure (PAP) and thereby prevent end-stage cardiopulmonary failure, have been used during recent years. Endothelin-1 has been found to be involved in the pathogenesis of PH. The dual endothelin-receptor antagonist, bosentan, was recently approved for the treatment of pulmonary arterial hypertension. The drug is mainly cleared by hepatic elimination. Severe renal dysfunction does not affect the single-dose pharmacokinetics of bosentan to a clinically relevant extent. Whether renal replacement therapy, however, interferes with the pharmacokinetics of bosentan is unknown. The authors report on the use of bosentan (125 mg twice daily) and its pharmacokinetic monitoring in a 19-year-old woman with PH and end-stage renal disease secondary to scleroderma. Treatment was well tolerated without drug specific adverse effects. After 12 months of treatment, pulmonary arterial pressure had normalized (48 mm Hg before start of treatment, 27 mm Hg at last follow-up). On the basis of analyzing samples from Genius-hemodialysis by a liquid chromatography assay with tandem mass spectrometry detection, the authors determined the bosentan dialysis clearance to be as low as 3.5 mL/min. Bosentan for the treatment of secondary PH seems to be safe as well as effective in end stage renal disease patients and no adjustment of the bosentan dosing regimen appears necessary. PMID- 15112185 TI - Three diabetic peritoneal dialysis patients receiving intraperitoneal insulin with dosage adjustment based on capillary glucose levels during peritoneal equilibrium tests. AB - Diabetic patients receiving peritoneal dialysis have vast differences in glucose absorption, significantly affecting glycemic control. Measurement of glucose levels between dialysate exchanges does not adequately reflect these changes. By measuring glucose levels during the peak period of glucose absorption, as determined by a peritoneal equilibrium test, such differences can be measured and accounted for when determining doses of intraperitoneal insulin. In this report, 3 cases in which such an approach was applied are described. PMID- 15112186 TI - Empowerment of patient preference in dialysis modality selection. PMID- 15112187 TI - Unexplained nephrotic-range proteinuria in a 38-year-old man: a case of "no change disease". PMID- 15112188 TI - Screening to prevent coronary events or screening to detect obstruction? PMID- 15112190 TI - Quiz page. Primary hyperparathyroidism. PMID- 15112192 TI - Collapsing glomerulopathy in adult still's disease. AB - Idiopathic collapsing glomerulopathy is a clinically and pathologically distinct variant of focal segmental glomerulosclerosis characterized clinically by a male and Afro-Caribbean racial predominance, proteinuria (often nephrotic range), and rapid progression to end-stage renal failure. Pathologically, the typical changes are global glomerular collapse leading to obliteration of glomerular capillary lumina, hypertrophy and hyperplasia of podocytes, and severe tubulointerstitial changes. A secondary form with almost identical pathologic features is described in association with human immunodeficiency virus infection. We describe a female patient who presented with multisystemic manifestations, including high spiking fever, arthralgias, lymphadenopathy, striking hyperferritinemia, and impaired renal function with proteinuria. Renal biopsy showed classic collapsing glomerulopathy. A diagnosis of adult Still's disease was made on the basis of Yamaguchi's criteria. The patient was treated with steroids, resulting in remission of the rheumatological condition closely paralleled by remission of proteinuria and renal function, thereby strongly suggesting a causative link between adult Still's disease and collapsing glomerulopathy in this patient. We propose that collapsing glomerulopathy ought to be considered in adult Still's disease with unexplained renal insufficiency or proteinuria. PMID- 15112193 TI - Shunt nephritis with positive titers for ANCA specific for proteinase 3. AB - The authors report a case of shunt nephritis with antineutrophil cytoplasmic autoantibody (ANCA) and review 2 similar cases. A 55-year-old man was admitted to our hospital for continuous fever and foot edema in 2002. A ventriculoperitoneal shunt was implanted because of a brain abscess and subsequent hydrocephalus in 1987; it was changed to a ventriculoatrial (VA) shunt in 1995. Urinary analysis showed proteinuria (5.4 g/d) and microscopic hematuria. Laboratory data showed renal dysfunction and hypocomplementemia. ANCA specific for proteinase 3 (PR3 ANCA) was positive in his serum, and blood culture grew Propionibacterium acnes. Renal biopsy results showed membranoproliferative glomerulonephritis type I. Therefore, the VA shunt was replaced, and antibiotics were administered. Oral prednisolone was initiated at a dose of 50 mg/d. Proteinuria and the serum levels of creatinine were improved concomitant with normalization of the serum complement levels and the decrease in serum PR3-ANCA titer. Similarly, another 2 cases reported in the literature of PR3-ANCA-positive shunt nephritis caused by P acnes and Gemella morbillorum showed good outcomes after removal of the shunt and administration of antibiotics with or without steroid therapy. PMID- 15112194 TI - Arteriovenous fistula-associated high-output cardiac failure: a review of mechanisms. AB - High-output cardiac failure can be a rare complication of high-output arteriovenous fistula. The authors present a case in which a hemodialysis patient with a high-flow arteriovenous fistula has cardiac failure that improves with fistula closure. The hemodynamic effects of a fistula are reviewed, and the hemodialysis literature regarding high-output cardiac failure is summarized. To gain insight into the problem of high-output cardiac failure, research efforts should focus on the prospective monitoring of high-access flows. PMID- 15112195 TI - A patient with severe renal amyloidosis associated with an immunoglobulin gamma heavy chain fragment. AB - The authors report a patient with progressive renal dysfunction caused by severe renal amyloidosis associated with a gamma-heavy chain variable region (V(H)) fragment. The patient was a 71-year-old woman who had renal insufficiency without nephrotic syndrome. Laboratory data showed a monoclonal IgG lambda component in her serum and urine. Renal biopsy results showed massive amyloid deposition in the mesangial region, but the glomerular basement membranes and epithelial cells were preserved. Because immunohistochemical studies using antibodies against a number of known amyloid fibril proteins failed to detect the amyloid protein, the amyloid protein extracted from a small piece of the biopsied renal tissue was subjected to biochemical analysis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of the extracted amyloid protein showed a prominent band at 11-kDa, and this protein was identified by amino acid sequence analysis as a gamma-heavy chain variable region fragment (V(H)3 subgroup) without the first N terminal residue. Our results indicate that the patient's renal amyloidosis was associated with a gamma-heavy chain variable region fragment. Microextraction and biochemical characterization of amyloid fibrils was of great use for reaching a definitive diagnosis. PMID- 15112196 TI - Transient visual loss may anticipate occipital infarction from hemodialysis. AB - BACKGROUND: The aim of this study was to report on a patient who had occipital infarction from intradialytic hypotension and review the literature on hemodialysis and visual loss. METHODS: Neuro-ophthalmologic examination and magnetic resonance imaging (MRI) of the brain were conducted. RESULTS: A 61-year old woman had acute onset of a right homonymous superior quadrantic visual field defect during dialysis after several episodes of transient visual loss associated with symptomatic intradialytic hypotension. MRI showed acute infarction in the left occipital lobe. CONCLUSION: Visual loss is an uncommon complication of hemodialysis. The mechanism usually is related to hypotension and is usually from anterior ischemic optic neuropathy. Although cortical blindness secondary to stroke has been reported as a complication after hemodialysis, we were unable to find another report of visual loss secondary to stroke attributed to intradialytic hypotension with preceding episodes of transient visual loss. Transient visual loss may be a risk factor for stroke during hemodialysis. PMID- 15112197 TI - Anti-CD20 monoclonal antibody (rituximab) treatment for hepatitis C-negative therapy-resistant essential mixed cryoglobulinemia with renal and cardiac failure. AB - A case of hepatitis C virus (HCV)-negative essential mixed cryoglobulinemia with initial skin, joint, and liver involvement, complicated by severe renal and cardiac failure and resistant to corticosteroid therapy, plasmapheresis, chlorambucil, and cyclophosphamide is presented. Treatment with an anti-CD20 monoclonal antibody (rituximab) induced a persistent recovery of renal and cardiac function, disappearance of skin and joint lesions, and disappearance of cryoglobulins. In a 24-month follow-up period, our patient remains in remission and free of symptoms. PMID- 15112199 TI - Powering nanodevices with biomolecular motors. AB - Biomolecular motors, in particular motor proteins, are ideally suited to introduce chemically powered movement of selected components into devices engineered at the micro- and nanoscale level. The design of such hybrid "bio/nano"-devices requires suitable synthetic environments, and the identification of unique applications. We discuss current approaches to utilize active transport and actuation on a molecular scale, and we give an outlook to the future. PMID- 15112200 TI - The nature of intermolecular CuI...CuI interactions: a combined theoretical and structural database analysis. AB - The nature of intermolecular interactions between dicoordinate Cu(I) ions is analyzed by means of combined theoretical and structural database studies. Energetically stable Cu(I).Cu(I) interactions are only found when the two monomers involved in the interaction are neutral or carry opposite charges, thus allowing us to speak of bonding between the components of the bimolecular aggregate. A perturbative evaluation of the components of the intermolecular interaction energies, by means the IMPT scheme of Stone, indicates that both the Coulombic and dispersion forces are important in determining the Cu(I).Cu(I) bonding interactions, because only a small part of that energy is attributable to Cu.Cu interactions, while a large component results from Cu.ligand interactions. The electrostatic component is the dominant one by far in the interaction between charged monomers, while in the interaction between neutral complexes, the electrostatic component is found to be of the same order of magnitude as the dispersion term. Bimolecular aggregates that have like charges are repulsive by themselves, and their presence in the solid state results from anion.cation interactions with ions external to this aggregate. In these cases, the short contact Cu.Cu interactions here should be more properly called counterion mediated Cu.Cu bonds. PMID- 15112201 TI - Catalytic asymmetric bromination and chlorination of beta-ketoesters. AB - The first general catalytic asymmetric bromination and chlorination of beta ketoesters has been developed. The reactions proceed for both acyclic and cyclic beta-ketoesters catalyzed by chiral bisoxazolinecopper(II) complexes giving the corresponding optically active alpha-bromo- and alpha-chloro-beta-ketoesters in high yields and moderate to good enantioselectivities. For the optically active chlorinated products the isolated yields are in the range of 88-99 % and the enantiomeric excesses up to 77 % ee, while the optically active brominated adducts are formed in 70-99 % isolated yield and up to 82 % ee. Based on the absolute configuration of the optically active products, the face selectivity for the catalytic enantioselective halogenation is discussed based on a bidentate coordination of the beta-ketoester to the chiral catalyst and a X-ray structure of chiral alpha,gamma-diketoesterenolatebisoxazolinecopper(II) complex. PMID- 15112202 TI - Sterically and guest-controlled self-assembly of calix[4]arene derivatives. AB - In solvents such as chloroform or benzene, tetraurea calix[4]arenes 1 form dimeric capsules in which one solvent molecule is usually included as guest. To explore the structural requirements for the formation of such hydrogen-bonded dimers we replaced one p-tolylurea residue by a simple acetamide function. The resulting calix[4]arene 2 a, substituted at its wide rim with one acetamide and three p-tolylurea functions, assumes a C(1)-symmetrical conformation in apolar solvents as shown by (1)H NMR, which is not compatible with the usual capsule. In the crystalline state, four molecules of 2 a, adopting a pinched cone conformation, assemble into a quasi S(4)-symmetrical tetramer stabilized by a cyclic array of 24 NH.O==C hydrogen bonds and four NH.pi interactions. Four acetamide groups are hydrogen-bonded to each other and pack tightly in the center of the assembly. All polar residues are buried inside the tetramer, the surface of which is lipophilic. Extensive NMR studies revealed similar structures in apolar solvents such as [D]chloroform or [D(6)]benzene for calixacetamides 2 a-c. The formation of these tetramers in solution is critically dependent on the size of the amide fragment, so that propionamide 2 d, butyramide 2 e, and p-tolylamide 2 f form only ill-defined aggregates. This is caused by steric crowding inside the tetrameric assembly. The tetramers persist during molecular dynamics simulations, and the optimized average structure of the MD run is similar to that found in the crystalline state. Theoretical studies revealed that cooperation of hydrogen bonds with multiple NH.pi, C--H.pi, and pi.pi attractions make the tetramer more stable than the capsular dimer with the solvent as guest. In the presence of tetraethylammonium salts, however, compounds 2 a-e form dimeric capsular assemblies, each incorporating a single ammonium cation. Only one of two possible regioisomeric dimers is formed, in which both acetamide groups are surrounded by two urea residues. These examples give striking evidence of how self-assembly in solution can be strongly dependent on subtle structural factors and of how the formation of dimeric capsules can be induced by the presence of an appropriate guest. PMID- 15112203 TI - Total synthesis of 34-hydroxyasimicin and its photoactive derivative for affinity labeling of the mitochondrial complex I. AB - The asymmetric total synthesis of the 34-hydroxyasimicin and its 3-(4 benzoylphenyl)propionate ester was achieved by means of a convergent synthetic strategy. This ester, which contains eight asymmetric centers, represents the first photoaffinity-labeling agent that is derived from an Annonaceous acetogenin. The key transformations in the synthesis include the Sharpless asymmetric dihydroxylation reaction, the Wittig olefination reaction, an oxidative cyclization reaction with rhenium(vii) oxide, the Williamson etherification reaction, and a palladium-catalyzed cross-coupling reaction. Use of the target molecule for photoaffinity-labeling studies of bovine mitochondrial NADH-ubiquinone oxidoreductase (Complex I) may shed light on the structure/function of this intricate enzyme and on the origin of the high antitumor activity exhibited by the Annonaceous acetogenins. PMID- 15112204 TI - Antibody-catalyzed benzoin oxidation as a mechanistic probe for nucleophilic catalysis by an active site lysine. AB - Aldolase antibody 24H6, which was obtained by reactive immunization against a 1,3 diketone hapten, is shown to catalyze additional reactions, including H/D exchange and oxidation reactions. Comparison of the H/D exchange reaction at the alpha-position of a wide range of aldehydes and ketones by 24H6 and by other aldolase antibodies, such as 38C2, pointed at the significantly larger size of the 24H6 active site. This property allowed for the catalysis of the oxidation of substituted benzoins to benzils by potassium ferricyanide. This reaction was used as a mechanistic probe to learn about the initial steps of the 24H6-catalyzed aldol condensation reaction. The Hammett correlation (rho=4.7) of log(k(cat)) versus the substituent constant, sigma, revealed that the reaction involves rapid formation of a Schiff base intermediate from the ketone and an active site lysine residue. The rate-limiting step in this oxidation reaction is the conversion of the Schiff base to an enamine intermediate. In addition, linear correlation (rho=3.13) was found between log(K(M)) and sigma, indicating that electronic rather than steric factors are dominant in the antibody-substrate binding phenomenon and confirming that the reversible formation of a Schiff base intermediate comprises part of the substrate-binding mechanism. PMID- 15112205 TI - Synthesis and redox behavior of ruthenocene-terminated oligoenes: characteristic and stable two-electron redox system and lower potential shift of the two electron oxidation wave with elongating conjugation. AB - Ruthenocene-terminated butadienes and hexatrienes were prepared by the Wittig reaction of 3-ruthenocenyl-2-propenals with ruthenocenylmethylphosphonium salts and the Mukaiyama coupling of the propenals, respectively. Cyclic voltammetry of these complexes indicated that they were involved in a stable two-electron redox process. The oxidation potentials for ruthenocene-terminated oligoenes shifted progressively to lower potential with the increasing CH==CH units as follows: Rc- Rc (0.32 V)>RcCH==CHRc (+0.09 V)>Rc(CH==CH)(2)Rc (-0.06 V)>Rc(CH==CH)(3)Rc (-0.07 V), (Rc=ruthenocene). The tendency is in remarkable contrast to that in the successive one-electron redox process. These complexes were chemically oxidized to give stable crystalline solids, whose structures were confirmed by NMR spectroscopy and X-ray analysis to be oligoene analogues of a bis(fulvene) complex, for example, [(eta(5)-C(5)Me(5))Ru[mu(2)-eta(6):eta(6) C(5)H(4)CH(CH==CH)(n)CHC(5)H(4)]Ru(eta(5)-C(5)Me(5))](2+) (n=1 or 2). The DFT calculation of the two-electron-oxidized species reproduced well the fulvene complex structure for the ruthenocene moieties. Since both the neutral and oxidized species are stable and chemically reversible, this redox system may be serviceable as a two-electron version of the ferrocene one-electron redox system. PMID- 15112206 TI - Enantioselective [4+2]-cycloaddition reaction of a photochemically generated o quinodimethane: mechanistic details, association studies, and pressure effects. AB - 1,2,3,4-Tetrahydro-2-oxoquinoline-5-aldehyde (2) was prepared from m-aminobenzoic acid and 3-ethoxyacryloyl chloride (4) in 19 % overall yield. Compound 2 underwent a photochemically induced [4+2]-cycloaddition reaction with various dienophiles upon irradiation in toluene solution. The exo product 10 a was obtained with acrylonitrile (9 a) as the dienophile, whereas methyl acrylate (9 b) and dimethyl fumarate (9 c) furnished the endo products 11 b and 11 c (69-77 % yield). The reactions proceeded at -60 degrees C in the presence of the chiral complexing agent 1 (1.2 equiv) with excellent enantioselectivity (91-94 % ee). The enantiomeric excess increases in the course of the photocycloaddition as a result of the lower product association to 1. The intermediate (E)-dienol 8 was spectroscopically detected at -196 degrees C in an EPA (diethyl ether/isopentane/ethanol) glass matrix. The association of the substrate 2 to the complexing agent 1 was studied by circular dichroism (CD) titration. The measured association constant (K(A)) was 589 M(-1) at room temperature (25 degrees C) and normal pressure (0.1 MPa). An increase in pressure led to an increased association. At 400 MPa the measured value of K(A) was 703 M(-1). Despite the stronger association the enantioselectivity of the reaction decreased with increasing pressure. At 25 degrees C the enantiomeric excess for the enantioselective reaction 2 + 9 a-->10 a decreased from 68 % ee at 0.1 MPa to 58 % ee at 350 MPa. This surprising behavior is explained by different activation volumes for the diastereomeric transition states leading to 10 a and ent-10 a. PMID- 15112207 TI - Group 1 coordination chains and hexagonal networks of host cyclotriveratrylene with halogenated monocarbaborane anions. AB - Halogenated carbaborane ions [CB(11)H(6)X(6)](-) in which X=Cl or Br have been combined with the host molecule cyclotriveratrylene (CTV) and Group 1 metal cations to give crystalline materials. The complexes [Na(ctv)(H(2)O)(CB(11)H(6)X(6))](CF(3)CH(2)OH) feature chiral Na-CTV coordination chains with complexation of the [CB(11)H(6)X(6)](-) ion by the Na(+) ion, together with the CTV molecular cavity. The coordination chains are hydrogen bonded together to give a puckered two-dimensional hexagonal grid structure. [K(ctv)(CB(11)H(6)Cl(6))(CF(3)CH(2)OH)(0.5)] is essentially isostructural. Complexes [Rb(ctv)(CB(11)H(6)Br(6))(H(2)O)] and [Cs(ctv)(CB(11)H(6)X(6))(CH(3)CN)] are coordination polymers with related distorted hexagonal grid structures. Use of N,N'-dimethylformamide (DMF) as a solvent results in an entirely different type of assembly, with [Na(2)(dmf)(4)(H(2)O)(2)(ctv)][(dmf)(0.5)(ctv)][CB(11)H(6)Br(6)](2) showing unusual [Na-mu-(dmf)-Na] bridges, and once again forming a distorted hexagonal coordination polymer. PMID- 15112208 TI - Surface-confined single molecules: assembly and disassembly of nanosize coordination cages on gold (111). AB - A cavitand functionalized with four alkylthioether groups at the lower rim, and four tolylpyridine groups on the upper rim is able to bind to a gold surface by its thioether groups, and forms a coordination cage with [Pd(dppp)(CF(3)SO(3))(2)] by its pyridine groups. The cavitand or the cage complex can be inserted from solution into a self-assembled monolayer (SAM) of 11 mercaptoundecanol on gold. The inserted molecules can be individually detected as they protrude from the SAM by atomic force microscopy (AFM). The cages can be reversibly assembled and disassembled on the gold surface. AFM can distinguish between single cavitand and cage molecules of 2.5 nm and 5.8 nm height, respectively. PMID- 15112209 TI - Rational design of the first closed coordination capsule with octahedral outer shape. AB - The rational synthesis of an octahedral coordination capsule in which the triangular faces are covered by single ligands is described herein. Starting with tris(2-hydroxybenzylidene)triaminoguanidinium chloride [H(6)L]Cl, we observed an oxidative cyclization of this ligand in the presence of PPh(4) (+) ions resulting in the complex [Pd(H(2)L')(PPh(3))] (1). The use of 5,5-diethylbarbiturate (bar(2 )) as a bridging ligand in the presence of [Co(en)(3)](3+) (en=ethylenediamine) leads to the formation of a rectangular box with the formula (Et(4)N)(6)[[Co[(PdCl)(Pd)L](2)(mu-bar)](2)] (2). The analysis of the architecture of compounds 1 and 2 enables the development of a self-assembly strategy for the synthesis of an octahedral coordination cage 3 with the formula Na(4)(Et(3)NH)(12)[(Pd(3)L)(8)[mu-(bar)](12)].x H(2)O. Compound 3 was characterized by (13)C-MAS-NMR spectroscopy and single-crystal structure analysis. PMID- 15112210 TI - Total synthesis of macroviracin D (BA-2836-4). AB - The first total synthesis of the complex glycolipid macroviracin D (BA-2836-4) (1) is described. This antivirally active metabolite isolated from the mycelium extracts of Streptomyces sp. BA-2836 incorporates a unique 46-membered macrodilactone motif decorated with glycosylated fatty acid appendices. Compound 1 consists of three identical subunits which are closely related to one of the segments found in cycloviracin B(1) (2), another antiviral glycoconjugate previously synthesized in our laboratory. Key steps of the synthesis route to 1 involve the stereoselective, ligand-controlled addition of the functionalized diorganozinc derivative 9 to aldehydes 8 a, b, a series of beta-selective glycosidation reactions using appropriately protected trichloroacetimidate donors, and three esterifications via the Yamaguchi method; one of them is performed intramolecularly to forge the macrocyclic lactone ring of the target in 89 % isolated yield. This total synthesis also firmly establishes the absolute configuration of the subunits of compound 1 as 3R,17S,23R. PMID- 15112211 TI - Computational studies of carbon nanotube-hydrocarbon bond strengths at nanotube ends: effect of link heteroatom and hydrocarbon structure. AB - Semiempirical and density functional electronic structure theory methods were used to study SWNT-X--R bond strengths, where the single-walled carbon nanotube (SWNT) had an armchair or zigzag structure, the link heteroatom X was O, N(H), or S and the hydrocarbon chain R was CH(2)CH(3), CH(OH)CH(3), CHCH(2), or CH(CF(3))CH(3). In all systems the hydrocarbon was bonded to the end of the nanotube. The SWNT-X--R bond (that is, the bond joining the link atom to the hydrocarbon) is more than 0.4 eV stronger for armchair than for zigzag nanotubes with the same diameters, irrespective of whether O, N, or S are used as link atoms or whether OH, C==C, or CF(3) groups are present in the hydrocarbon chain. This raises the possibility for selective manipulation of armchair/zigzag nanotubes using a variety of link atoms and hydrocarbon structures. The SWNT-O- CH(CF(3))CH(3) bond is weaker than the SWNT-O--CH(2)CH(3) bond (for both armchair and zigzag nanotubes), while inclusion of a double bond in the ethyl chain increases the bond strengths. Also, SWNT-S--CH(2)CH(3) and SWNT-N(H)--CH(2)CH(3) bonds are stronger than SWNT-O--CH(2)CH(3) bonds. PMID- 15112212 TI - Cuprophilic interactions in luminescent copper(I) clusters with bridging bis(dicyclohexylphosphino)methane and iodide ligands: spectroscopic and structural investigations. AB - Polynuclear copper(I) complexes with bridging bis(dicyclohexylphosphino)methane (dcpm) and iodide ligands, [Cu(2)(dcpm)(2)(CH(3)CN)(2)](BF(4))(2) (1), [Cu(2)(dcpm)(2)](BF(4))(2) (2), [(CuI)(3)(dcpm)(2)] (3), [(CuI)(4)(dcpm)(2)] (4), and [(CuI)(2)(dcpm)(2)] (5) were prepared and their structures determined by X ray crystal analysis. The shortest Cu--Cu distance found in these complexes is 2.475(1) A for 3. Powdered samples of 1, 3, 4, and 5 display intense and long lived phosphorescence with lambda(max) at 460, 626, 590, and 456 nm and emission quantum yields of 0.26, 0.11, 0.12, and 0.56 at room temperature, respectively. In the solid state, 2 displays both a weak emission at 377 and an intense one at 474 nm with an overall emission yield 0.42. The difference in emission properties among complexes 1-5 suggests that both Cu--Cu interaction and coordination around the copper(I) center affect the excited state properties. A degassed solution of 2 in acetone gives a bright red emission with lambda(max) at 625 nm at room temperature. The difference absorption spectra of the triplet excited states of 1 5 in acetonitrile show broad absorption peaks at 340-410 and 850-870 nm. PMID- 15112213 TI - A "chiral aldehyde" equivalent as a building block towards biologically active targets. AB - Chiral gamma-aryloxybutenolides, readily accessible through dynamic kinetic asymmetric transformation (DYKAT) of racemic acyloxybutenolides, were utilized as "chiral aldehyde" building blocks for intermolecular cycloadditions and Michael reactions. Unprecedented selectivity in trimethylenemethane cycloadditions with this building block allowed an efficient synthesis of a novel metabotropic glutamate receptor 1 antagonist in development by the Bayer corporation. These studies further inspired work that culminated in the total synthesis of (+) brefeldin A, a natural product with a range of significant biological properties. All of the stereochemistry in this target molecule was derived from two palladium catalyzed asymmetric allylic alkylation reactions. The trans-alkenes were synthesized by a Julia olefination and a ruthenium-catalyzed trans hydrosilylation-protodesilylation protocol. The route to (+)-brefeldin A lends itself to analogue syntheses and was completed in 18 steps in 6 % overall yield. PMID- 15112214 TI - Excited-state behavior and photoionization of 1,8-acridinedione dyes in micelles- comparison with NADH oxidation. AB - The photophysics and photochemistry of 1,8-acridinedione dyes, which are analogues of reduced nicotinamide adenine dinucleotide (NADH), are studied in anionic and cationic micelles. Acridinedione dyes (ADDs) are solubilized in micelles at the micelle-water interface and are in equilibrium between the aqueous and micellar phase. The binding of the ADDs with micelles is attributed to hydrophobic interactions and the binding constants are determined with steady state and time-resolved techniques. Nanosecond laser flash photolysis studies are carried out in aqueous, anionic, and cationic micellar solutions. The ADD undergoes photoionization in the excited state to give a solvated electron. The solvated electron reacts with the ADD to give an anion radical. In anionic micelles, the yield of the solvated electron increases because of the efficient separation of the cation radical and the electron. Cation radicals derived from the photooxidation of ADDs are involved in keto-enol tautomerization. Under acidic conditions, an enol radical cation of the acridinedione dye is formed from the keto form of the cation radical by intramolecular hydrogen atom transfer. In cationic micelles, due to electrostatic attraction, the electron cannot escape from the micelle and recombination of the cation radical and the electron results in the formation of a triplet state. For the first time, a solvated electron is observed in the laser flash photolysis of ADDs in anionic micelles. The photoionization of ADDs depends on the excitation wavelength and is biphotonic at 355 nm and monophotonic at 248 nm. From the results with this NADH model compound, the sequential electron-proton-electron transfer oxidation of NADH is confirmed and the nature of the intermediates involved in the oxidation is unraveled; these intermediates are found to depend on the pH value of the medium. PMID- 15112215 TI - Metallosupramolecular zippers generated by self-organization of self complementary molecular clefts. AB - The binding of Co(2+) and Pb(2+) ions to the terpyridine and pyridine subunits of the ligand 1 leads to the self-complementary molecular clefts 2-6, which result from the crossover combination of orthogonal-terpyridine and linear-pyridine metal-coordination subprograms and are stabilized by strong pi-pi stacking interactions. Four different cleft-type entities, [Co(2+) (2)(1)(2)] (3), [Pb(2+) (2)(1)(2)] (4), [Co(2+) (4)(1)(2)] (5), [Pb(2+) (4)(1)(2)] (6), are generated in both solution and the solid state, and may be interconverted as a function of metal/ligand stoichiometry. One- and two-dimensional metallosupramolecular zipper architectures result from self-assembly in the solid state driven by a combination of different pi-pi stacking subprograms. The U-shaped geometry of the ligand influences the possibility of zipping and thus, in turn, the generation of different zipper architectures. The structures of 2-5 have been confirmed by X ray crystallography; that of 6 is based on NMR spectral data. PMID- 15112216 TI - thermodynamic/kinetic control in the isomerization of the [[tBuNP(mu-NtBu)]2]2- ion. AB - The unique structure of [(tBuN)(2)PK]( infinity ) (2) (containing [(tBuN)(2)P](-) monoanions) is in stark contrast to the previously reported Li(+) analogue [[[tBuNP(mu-NtBu)](2)](2)]Li(4) (1) (containing the dimeric [[tBuNP(mu NtBu)](2)](2-) ion). DFT and (31)P NMR spectroscopic studies reveal that the formation of the monoanion arrangements are most thermodyamically favored for Li, Na, and K, 1 being the product of kinetic control and 2 being the product of thermodynamic control. PMID- 15112217 TI - Ultra-deep desulfurization of diesel: oxidation with a recoverable catalyst assembled in emulsion. AB - A [(C(18)H(37))(2)N(+)(CH(3))(2)](3)[PW(12)O(40)] catalyst, assembled in an emulsion in diesel, can selectively oxidize the sulfur-containing molecules present in diesel into their corresponding sulfones by using H(2)O(2) as the oxidant under mild conditions. The sulfones can be readily separated from the diesel using an extractant, and the sulfur level of the desulfurized diesel can be lowered from about 500 ppm to 0.1 ppm without changing the properties of the diesel. The catalyst demonstrates high performance (>/=96 % efficiency of H(2)O(2), is easily recycled, and approximately 100 % selectivity to sulfones). Metastable emulsion droplets (water in oil) act like a homogeneous catalyst and are formed when the catalyst (as the surfactant) and H(2)O(2) (30 %) are mixed in the diesel. However, the catalyst can be separated from the diesel after demulsification. PMID- 15112218 TI - Metmyoglobin-catalyzed exogenous and endogenous tyrosine nitration by nitrite and hydrogen peroxide. AB - Metmyoglobin catalyzes the nitration of various phenolic compounds in the presence of nitrite and hydrogen peroxide. The reaction rate depends on the reactant concentrations and shows saturation behavior. Two competing paths are responsible for the reaction. In the first, myoglobin reacts according to a peroxidase-like cycle forming two active intermediates, which can induce one electron oxidation of the substrates. The MbFe(IV)==O intermediate oxidizes nitrite to nitrogen dioxide, which, after reaction with the phenol or with a phenoxy radical, yields the nitrophenol. In the second mechanism, hydrogen peroxide reacts with iron-bound nitrite to produce an active nitrating species, which we assume to be a protein-bound peroxynitrite species, MbFe(III)--N(O)OO. The high nitrating power of the active species is shown by the fact that the catalytic rate constant is essentially independent of the redox properties of the phenol. The occurrence of one or other of these mechanisms depends on the nitrite concentration: at low [NO(2) (-)] the nitrating agent is nitrogen dioxide, whereas at high [NO(2) (-)] the peroxynitrite path is dominant. The myoglobin derivative that accumulates during turnover depends on the mechanism. When the path involving NO(2) (.) is dominant, the spectrum of the MbFe(IV)==O intermediate is observed. At high nitrite concentration, the Soret band appears at 416 nm, which we attribute to an iron-peroxynitrite species. The metMb/NO(2) ( )/H(2)O(2) system competitively nitrates the heme and the endogenous tyrosine at position 146 of the protein. Phenolic substrates protect Tyr146 from nitration by scavenging the active nitrating species. The exposed Tyr103 residue is not nitrated under the same conditions. PMID- 15112219 TI - Mechanism of nitrosylmyoglobin autoxidation: temperature and oxygen pressure effects on the two consecutive reactions. AB - As shown by singular value decomposition and global analysis of the absorption spectra, oxidation of nitrosylmyoglobin, MbFe(II)NO, by oxygen occurs in two consecutive (pseudo) first-order reactions in aqueous air- saturated solutions at physiological conditions (pH 7.0, I=0.16 m (NaCl)). Both reaction steps have a large temperature dependence with the following activation parameters: DeltaS++(1) = 121+/-7 and DeltaS++(1) = 23+/-29; and DeltaS++(2) = 88+/-14 kJ mol(-1) and DeltaS++(2)-63+/-51 J(-1) K(-1) mol(-1) at 25 degrees C for the first and second step, respectively. At physiological temperature, the initial reaction is faster, while at lower temperatures, the first reaction is slower and rate determining. The rate of the first reaction is linearly dependent on oxygen pressure at lower pressures, while for oxygen pressures above atmospheric, the rate exhibits saturation behaviour. The second reaction is independent of oxygen pressure. The rate of the second reaction increases when oxymyoglobin is added. In contrast, the rate of the first reaction is independent of the presence of oxymyoglobin. The observed kinetics are in agreement with a reaction mechanism in which the nitric oxide that is initially bound to the Fe(II) centre of myoglobin is displaced by oxygen in a reversible ligand-exchange reaction prior to an irreversible electron transfer. The ligand-exchange process is dissociative in nature and depends bond breaking, and nitric oxide is suggested to be trapped in a protein cavity. The absorption spectrum of the intermediate, as resolved from the global analysis, is in agreement with a peroxynitrite complex, and the initial process must involve partial electron transfer. PMID- 15112220 TI - N,N'-ethylenebis(pyridoxylideneiminato) and N,N'-ethylenebis(pyridoxylaminato): synthesis, characterization, potentiometric, spectroscopic, and DFT studies of their vanadium(IV) and vanadium(V) complexes. AB - The Schiff base N,N'-ethylenebis(pyridoxylideneiminato) (H(2)pyr(2)en, 1) was synthesized by reaction of pyridoxal with ethylenediamine; reduction of H(2)pyr(2)en with NaBH(4) yielded the reduced Schiff base N,N' ethylenebis(pyridoxylaminato) (H(2)Rpyr(2)en, 2); their crystal structures were determined by X-ray diffraction. The totally protonated forms of 1 and 2 correspond to H(6)L(4+), and all protonation constants were determined by pH potentiometric and (1)H NMR titrations. Several vanadium(IV) and vanadium(V) complexes of these and other related ligands were prepared and characterized in solution and in the solid state. The X-ray crystal structure of [V(V)O(2)(HRpyr(2)en)] shows the metal in a distorted octahedral geometry, with the ligand coordinated through the N-amine and O-phenolato moieties, with one of the pyridine-N atoms protonated. Crystals of [(V(V)O(2))(2)(pyren)(2)].2 H(2)O were obtained from solutions containing H(2)pyr(2)en and oxovanadium(IV), where Hpyren is the "half" Schiff base of pyridoxal and ethylenediamine. The complexation of V(IV)O(2+) and V(V)O(2) (+) with H(2)pyr(2)en, H(2)Rpyr(2)en and pyridoxamine in aqueous solution were studied by pH-potentiometry, UV/Vis absorption spectrophotometry, as well as by EPR spectroscopy for the V(IV)O systems and (1)H and (51)V NMR spectroscopy for the V(V)O(2) systems. Very significant differences in the metal-binding abilities of the ligands were found. Both 1 and 2 act as tetradentate ligands. H(2)Rpyr(2)en is stable to hydrolysis and several isomers form in solution, namely cis-trans type complexes with V(IV)O, and alpha-cis- and beta-cis-type complexes with V(V)O(2). The pyridinium N atoms of the pyridoxal rings do not take part in the coordination but are involved in acid-base reactions that affect the number, type, and relative amount of the isomers of the V(IV)O-H(2)Rpyr(2)en and V(V)O(2)-H(2)Rpyr(2)en complexes present in solution. DFT calculations were carried out and support the formation and identification of the isomers detected by EPR or NMR spectroscopy, and the strong equatorial and axial binding of the O-phenolato in V(IV)O and V(V)O(2) complexes. Moreover, the DFT calculations done for the [V(IV)O(H(2)Rpyr(2)en)] system indicate that for almost all complexes the presence of a sixth equatorial or axial H(2)O ligand leads to much more stable compounds. PMID- 15112221 TI - Synthesis and characterisation of nu3-octahedral [Ni36Pd8(CO)48]6- and [Ni35Pt9(CO)48]6- clusters displaying unexpected surface segregation of Pt atoms and molecular and/or crystal substitutional Ni/Pd and Ni/Pt disorder. AB - The synthesis and structure, as well as the chemical and electrochemical characterisation of two new nu(3)-octahedral bimetallic clusters with the general [Ni(44-x)M(x)(CO)(48)](6-) (M = Pd, x = 8; M = Pt, x = 9) formula is reported. The [Ni(35)Pt(9)(CO)(48)](6-) cluster was obtained in reasonable yields (56 % based on Pt) by reaction of [Ni(6)(CO)(12)](2-) with 1.1 equivalents of Pt(II) complexes, in ethyl acetate or THF as the solvent. The [Ni(36)Pd(8)(CO)(48)](6-) cluster was obtained from the related reaction with Pd(II) salts in THF, and was isolated only in low yields (5-10 % based on Pd), mainly because of insufficient differential solubility of its salts. The unit cell of the [NBu(4)](6)[Ni(35)Pt(9)(CO)(48)] salt contains a substitutionally Ni-Pt disordered [Ni(24)(Ni(14-x)Pt(x))Pt(6)(CO)(48)](6-) (x = 3) hexaanion. A combination of crystal and molecular disorder is necessary to explain the disordering observed for the Ni/Pt sites. The unit cell of the corresponding [Ni(36)Pd(8)(CO)(48)](6-) salt contains two independent [Ni(30)(Ni(8 x)Pd(x))Pd(6)(CO)(48)](6-) (x = 2) hexaanions. The two display similar substitutional Ni-Pd disorder, which probably arises only from crystal disorder. The structure of [Ni(36)Pd(8)(CO)(48)](6-) establishes the first similarity between the chemistry of Ni-Pd and Ni-Pt carbonyl clusters. A comparison of the chemical and electrochemical properties of [Ni(35)Pt(9)(CO)(48)](6-) with those of the related [Ni(38)Pt(6)(CO)(48)](6-) cluster shows that surface colouring of the latter with Pt atoms decreases redox as well as protonation propensity of the cluster. In contrast, substitution of all internal Pt and two surface Ni with Pd atoms preserves the protonation behaviour and is only detrimental with respect to its redox aptitude. A qualitative rationalisation of the different surface-site selectivity of Pt and Pd, based on distinctive interplays of M--M and M--CO bond energies, is suggested. PMID- 15112222 TI - The photoisomerization mechanism of azobenzene: a semiclassical simulation of nonadiabatic dynamics. AB - We have simulated the photoisomerization dynamics of azobenzene, taking into account internal conversion and geometrical relaxation processes, by means of a semiclassical surface hopping approach. Both n-->pi* and pi-->pi* excitations and both cis-->trans and trans-->cis conversions have been considered. We show that in all cases the torsion around the N==N double bond is the preferred mechanism. The quantum yields measured are correctly reproduced and the observed differences are explained as a result of the competition between the inertia of the torsional motion and the premature deactivation of the excited state. Recent time-resolved spectroscopic experiments are interpreted in the light of the simulated dynamics. PMID- 15112223 TI - Selective complexation and transport of europium ions at the interface of vesicles. AB - The aim of the present work was to design functionalized lipidic membranes that can selectively interact with lanthanide ions at the interface and to exploit the interaction between membranes induced by this molecular-recognition process with a view to building up self-assembled vesicles or controlling the permeability of the membrane to lanthanide ions. Amphiphilic molecules bearing a beta-diketone unit as head group were synthesized and incorporated into phospholipidic vesicles. Binding of Eu(III) ions to the amphiphilic ligand can lead to formation of a complex involving ligands of the same vesicle membrane (intravesicular complex) or of two different vesicles (intervesicular complex). The effect of Eu(III) ions on vesicle behavior was studied by complementary techniques such as fluorimetry, light scattering, and electron microscopy. The formation of an intravesicular luminescent Eu/beta-diketone ligand (1/2) complex was demonstrated. The linear increase in the binding constant with increasing concentration of ligands in the membrane revealed a cooperative effect of the ligands distributed in the vesicle membrane. The luminescence of this complex can be exploited to monitor the kinetics of complexation at the interface of the vesicles, as well as ion transport across the membrane. By encapsulation of 2,6 dipicolinic acid (DPA) as a competing ligand which forms a luminescent Eu/DPA complex, the kinetics of ion transport across the membrane could be followed. These functional vesicles were shown to be an efficient system for the selective transport of Eu(III) ions across a membrane with assistance by beta-diketone ligands. PMID- 15112225 TI - Phylogeny of gamma-proteobacteria: resolution of one branch of the universal tree? AB - The reconstruction of bacterial evolutionary relationships has proven to be a daunting task because variable mutation rates and horizontal gene transfer (HGT) among species can cause grave incongruities between phylogenetic trees based on single genes. Recently, a highly robust phylogenetic tree was constructed for 13 gamma-proteobacteria using the combined alignments of 205 conserved orthologous proteins.1 Only two proteins had incongruent tree topologies, which were attributed to HGT between Pseudomonas species and Vibrio cholerae or enterics. While the evolutionary relationships among these species appears to be resolved, further analysis suggests that HGT events with other bacterial partners likely occurred; this alters the implicit assumption of gamma-proteobacteria monophyly. Thus, any thorough reconstruction of bacterial evolution must not only choose a suitable set of molecular markers but also strive to reduce potential bias in the selection of species. PMID- 15112226 TI - Prions at the crossroads: the need to identify the active TSE agent. AB - Structural change in the cellular prion protein, PrPC to a ProteinaseK-resistant beta-sheet-rich insoluble form PrPSC and its accumulation have been considered to be central to the pathogenesis of the prion diseases (TSE). In a recent paper, Deleault et al have shown that specific endogenous RNA molecules can induce in vitro structural conversion of endogenous PrPC to PrPSC. Small highly structured synthetic RNAs can also induce this conversion process. However, recent in vivo results show that PrPSC is not directly involved in the prion pathogenesis. It is possible, however, that nucleic-acid-induced PrPSC associated with the inducer nucleic acid could be the components of the infectious agent. PMID- 15112227 TI - The cnidarian and the canon: the role of Wnt/beta-catenin signaling in the evolution of metazoan embryos. AB - In a recent publication, Wikramanayake and colleagues have implicated the canonical Wnt/beta-catenin signaling pathway as a mediator of axial polarity and germ-layer specification in embryos of the cnidarian Nematostella. In this anthozoan, beta-catenin is localized in nuclei of blastomeres in one region of the 16- to 32-cell embryo whose descendants subsequently form the entoderm of the embryo. They claim that the pattern of nuclear localization is significant for two reasons: (1) when nuclear localization of beta-catenin was inhibited, gastrulation does not occur, and (2) when localization of beta-catenin took place in all cells of the pregastrula embryo, the number of entodermal cells increases. Since the Wnt/beta-catenin signaling pathway also plays a role in establishing axial polarity and specifying endoderm and mesoderm in a number of bilaterians, Wikramanayake et al. imply that this developmental mechanism is an evolutionary inheritance from a radially symmetrical ancestor. Some of the gaps in the current evidence, which must be filled to evaluate their interpretation, are discussed. PMID- 15112228 TI - The evolution of molecular genetic pathways and networks. AB - There is growing interest in the evolutionary dynamics of molecular genetic pathways and networks, and the extent to which the molecular evolution of a gene depends on its position within a pathway or network, as well as over-all network topology. Investigations on the relationships between network organization, topological architecture and evolutionary dynamics provide intriguing hints as to how networks evolve. Recent studies also suggest that genetic pathway and network structures may influence the action of evolutionary forces, and may play a role in maintaining phenotypic robustness in organisms. PMID- 15112229 TI - Can flies help humans treat neurodegenerative diseases? AB - Neurodegenerative diseases are becoming increasingly common as life expectancy increases. Recent years have seen tremendous progress in the identification of genes that cause these diseases. While mutations have been found and cellular processes defined that are altered in the disease state, the identification of treatments and cures has proven more elusive. The process of finding drugs and therapies to treat human diseases can be slow, expensive and frustrating. Can model organisms such as Drosophila speed the process of finding cures and treatments for human neurodegenerative diseases? We pose three questions, (1) can one mimic the essential features of human diseases in an organism like Drosophila, (2) can one cure a model organisms of human disease and (3) will these efforts accelerate the identification of useful therapies for testing in mice and ultimately humans? Here we focus on the use of Drosophila to identify potential treatments for neurodegenerative diseases such as Huntington's and we discuss how well these therapies translate into mammalian systems. PMID- 15112230 TI - The cadherin-catenin complex as a focal point of cell adhesion and signalling: new insights from three-dimensional structures. AB - Cadherins are a large family of single-pass transmembrane proteins principally involved in Ca2+-dependent homotypic cell adhesion. The cadherin molecules comprise three domains, the intracellular domain, the transmembrane domain and the extracellular domain, and form large complexes with a vast array of binding partners (including cadherin molecules of the same type in homophilic interactions and cellular protein catenins), orchestrating biologically essential extracellular and intracellular signalling processes. While current, contrasting models for classic cadherin homophilic interaction involve varying numbers of specific repeats found in the extracellular domain, the structure of the domain itself clearly remains the main determinant of cell stability and binding specificity. Through intracellular interactions, cadherin enhances its adhesive properties binding the cytoskeleton via cytoplasmic associated factors alpha- catenin, beta-catenin and p120ctn. Recent structural studies on classic cadherins and these catenin molecules have provided new insight into the essential mechanisms underlying cadherin-mediated cell interaction and catenin-mediated cellular signalling. Remarkable structural diversity has been observed in beta catenin recognition of other cellular factors including APC, Tcf and ICAT, proteins that contribute to or compete with cadherin/catenin functioning. PMID- 15112231 TI - Left and right in the amphibian world: which way to develop and where to turn? AB - The last decade has seen a dramatic increase in studies on the development, function and evolution of asymmetries in vertebrates, including amphibians. Here we discuss current knowledge of behavioral and anatomical asymmetries in amphibians. Behavioral laterality in the response of both adult and larval anurans to presumed predators and competitors is strong and may be related, respectively, to laterality in the telencephalon of adults and the Mauthner neurons of tadpoles. These behavior lateralities, however, do not seem to correlate with visceral asymmetries in the same animals. We briefly compare what is known about the evolution and development of asymmetry in the structure and function of amphibians with what is known about asymmetries in other chordate and non-chordate groups. Available data suggest that the majority of asymmetries in amphibians fall into two independent groups: (1) related to situs viscerum and (2) of a neurobehavioral nature. We find little evidence linking these two groups, which implies different developmental regulatory pathways and independent evolutionary histories for visceral and telencephalic lateralizations. Studies of animals other than standard model species are essential to test hypotheses about the evolution of laterality in amphibians and other chordates. PMID- 15112232 TI - Insulator dynamics and the setting of chromatin domains. AB - The early discovery of cis-regulatory elements able to promote transcription of genes over large distances led to the postulate that elements, termed insulators, should also exist that would limit the action of enhancers, LCRs and silencers to defined domains. Such insulators were indeed found during the past fifteen years in a wide range of organisms, from yeast to humans. Recent advances point to an important role of transcription factors in insulator activity and demonstrate that the operational observation of an insulator effect relies on a delicate balance between the "efficiency" of the insulator and that of the element to be counteracted. In addition, genuine insulator elements now appear less common than initially envisaged, and they are only found at loci displaying a high density of coding or regulatory information. Where this is not the case, chromatin domains of opposing properties are thought to confront each other at "fuzzy" boundaries. In this article, we propose models for both fixed and fuzzy boundaries that incorporate probabilistic and dynamic parameters. PMID- 15112233 TI - Factors contributing to the outcome of oxidative damage to nucleic acids. AB - Oxidative damage to DNA appears to be a factor in cancer, yet explanations for why highly elevated levels of such lesions do not always result in cancer remain elusive. Much of the genome is non-coding and lesions in these regions might be expected to have little biological effect, an inference supported by observations that there is preferential repair of coding sequences. RNA has an important coding function in protein synthesis, and yet the consequences of RNA oxidation are largely unknown. Some non-coding nucleic acid is functional, e.g. promoters, and damage to these sequences may well have biological consequences. Similarly, oxidative damage to DNA may promote microsatellite instability, inhibit methylation and accelerate telomere shortening. DNA repair appears pivotal to the maintenance of genome integrity, and genetic alterations in repair capacity, due to single nucleotide polymorphisms or mutation, may account for inter-individual differences in cancer susceptibility. This review will survey these aspects of oxidative damage to nucleic acids and their implication for disease. PMID- 15112234 TI - Battle of the Xs. AB - Females and males often exhibit conspicuous morphological, physiological and behavioral differences. Similarly, gene expression profiles indicate that a large portion of the genome is sex-differentially deployed, particularly in the germ line. Because males and females are so fundamentally different, each sex is likely to have a different optimal gene expression profile that is never fully achieved in either sex because of antagonistic selection in females versus males. Males are hemizygous for the X chromosome, which means that recessive male favorable de novo mutations on the X chromosome are subject to immediate selection. In females, a recessive female-favorable mutation on one of two X chromosomes is not available for selection until it becomes frequent enough in the local population to result in homozygous individuals. Given that most mutations are recessive, one would expect that genes or alleles favoring males should accumulate on the X chromosome. Recent microarray work in Drosophila and C. elegans clearly shows the opposite. Why is the X chromosome a highly disfavored location for genes with male-biased expression in these animals? PMID- 15112235 TI - Ion condensation and signal transduction. AB - Many abiotic and other signals are transduced in eukaryotic cells by changes in the level of free calcium via pumps, channels and stores. We suggest here that ion condensation should also be taken into account. Calcium, like other counterions, is condensed onto linear polymers at a critical value of the charge density. Such condensation resembles a phase transition and has a topological basis in that it is promoted by linear as opposed to spherical assemblies of charges. Condensed counterions are delocalised and can diffuse in the so-called near region along the polymers. It is generally admitted that cytoskeletal filaments, proteins colocalised with these filaments, protein filaments distinct from cytoskeletal filaments, and filamentous assemblies of other macromolecules, constitute an intracellular macromolecular network. Here we draw attention to the fact that this network has physicochemical characteristics that enable counterion condensation. We then propose a model in which the feedback relationships between the condensation/decondensation of calcium and the activation of calcium dependent kinases and phosphatases control the charge density of the filaments of the intracellular macromolecular network. We show how condensation might help mediate free levels of calcium both locally and globally. In this model, calcium condensation/decondensation on the macromolecular network creates coherent patterns of protein phosphorylation that integrate signals. This leads us to hypothesize that the process of ion condensation operates in signal transduction, that it can have an integrative role and that the macromolecular network serves as an integrative receptor. PMID- 15112236 TI - Endosymbiotic origins of sex. AB - Understanding how complex sexual reproduction arose, and why sexual organisms have been more successful than otherwise similar asexual organisms, is a longstanding problem in evolutionary biology. Within this problem, the potential role of endosymbionts or intracellular pathogens in mediating primitive genetic transfers is a continuing theme. In recent years, several remarkable activities of mitochondria have been observed in the germline cells of complex eukaryotes, and it has been found that bacterial endosymbionts related to mitochondria are capable of manipulating diverse aspects of metazoan gametogenesis. An attempt is made here to rationalize these observations with an endosymbiotic model for the evolutionary origins of sex. It is hypothesized that the contemporary life cycle of germline cells has descended from the life cycle of the endosymbiotic ancestor of the mitochondrion. Through an actin-based motility that drove it from one cell to another, the rickettsial ancestor of mitochondria may have functioned as a primitive transducing particle, the evolutionary progenitor of sperm. PMID- 15112237 TI - Insights into the evolution of the nucleolus by an analysis of its protein domain repertoire. AB - Recently, the first investigation of nucleoli using mass spectrometry led to the identification of 271 proteins. This represents a rich resource for a comprehensive investigation of nucleolus evolution. We applied a protocol for the identification of known and novel conserved protein domains of the nucleolus, resulting in the identification of 115 known and 91 novel domain profiles. The phyletic distribution of nucleolar protein domains in a collection of complete proteomes of selected organisms from all domains of life confirms the archaebacterial origin of the core machinery for ribosome maturation and assembly, but also reveals substantial eubacterial and eukaryotic contributions to nucleolus evolution. We predict that, in different phases of nucleolus evolution, protein domains with different biochemical functions were recruited to the nucleolus. We suggest a model for the late and continuous evolution of the nucleolus in early eukaryotes and argue against an endosymbiotic origin of the nucleolus and the nucleus. Supplementary material for this article can be found on the BioEssays website at http://www.interscience.wiley.com/jpages/0265 9247/suppmat/index.html. PMID- 15112238 TI - The ribosome: lifting the veil from a fascinating organelle. AB - It was first suggested that the ribosome is associated with protein synthesis in the 1950s. Initially, its components were revealed as surface-accessible proteins and as molecules of RNA apparently providing a scaffold for subunit shape. Attributing function to the proteins proved difficult, although bacterial protein L11 proved essential for binding one of the decoding protein release factors (RFs). With the discovery that RNA could be a catalyst, interest focussed on the rRNA that, in partnership with mRNA and tRNAs, could potentially mediate the chemical reaction underlying protein synthesis. rRNA interactions and conformational changes were invoked as key elements that facilitated function. The decoding RFs, which are proteins, are exceptions to this rule because they usurp a tRNA function in mediating stop signal recognition. Cryoelectron microscopy and associated image reconstruction technology have now given dramatic snapshots of almost every step of protein synthesis, and X-ray crystallography has revealed, at last, the subunits and monomeric ribosome in exquisite atomic detail. PMID- 15112239 TI - Pathogenic archaebacteria: do they not exist because archaebacteria use different vitamins? PMID- 15112241 TI - Abstracts of the XXII Congress of the International Society for Analytical Cytology, 22-27 May 2004. PMID- 15112245 TI - Squamous cell carcinoma of the anal margin. AB - PURPOSE: To define the optimal treatment of patients with squamous cell carcinoma (SCCA) of the anal margin. METHODS: Nineteen patients treated with curative intent by radiotherapy (RT) alone or combined with adjuvant chemotherapy (CTX) between 1979 and June 2000 were analyzed. The pertinent literature was reviewed and discussed as it related to our experience. RESULTS: Local control after RT or RT and CTX was observed in all 19 patients (100%). One T1 patient developed inguinal lymph node metastases and subsequently died secondary to regional and distant disease. This patient did not receive elective inguinal node RT; the lymph nodes of the other 18 patients in this analysis were irradiated. Four patients died of intercurrent disease at 25, 29, 37, and 113 months after RT, respectively. The remaining 14 patients were alive and disease-free from 52 to 143 months after treatment. No patient suffered a severe complication or required a diverting colostomy or an abdominoperineal resection (APR) after treatment. Review of the literature reveals that the probability of cure is similar after RT alone or combined with CTX compared with surgery. Therefore, the choice of treatment depends on the anticipated functional result. CONCLUSIONS: Patients with SCCA of the anal margin have a high likelihood of cure with sphincter preservation after RT or RT and CTX. Patients with well to moderately differentiated T1 tumors may undergo excision if it can be accomplished without compromising the sphincter. The remainder are treated with RT. Adjuvant CTX is indicated for those with T3-T4 tumors and/or involved regional nodes. PMID- 15112247 TI - Peritoneal carcinomatosis from colorectal or appendiceal origin: correlation of preoperative CT with intraoperative findings and evaluation of interobserver agreement. AB - BACKGROUND AND OBJECTIVES: In patients with colorectal cancer, it is important to diagnose peritoneal carcinomatosis as well as to detect location and size of peritoneal tumor dissemination in view of treatment planning. The aim of this study was to investigate the detection accuracy of computed tomography (CT). METHODS: Preoperative CT-scans from 25 consecutive patients with peritoneal carcinomatosis from colorectal or appendiceal origin were independently blindly reviewed by 2 radiologists. The presence and diameter of tumor deposits were noted in seven abdominopelvic areas. Intraoperative findings were regarded as the gold standard. Agreement was assessed using the Kappa index and the chi-square test. RESULTS: The presence of peritoneal carcinomatosis was detected in 60 and 76% of those patients by each of the radiologist. Detection of individual peritoneal implants was poor (kappa = 0.11/0.23) and varied from 9.1%/24.3% for tumor size <1 cm to 59.3%/66.7% for tumor size >5 cm. Overall sensitivity, specificity, accuracy, positive (PPV) and negative predictive value (NPV) for tumor involvement per area were 24.5%/37.3%, 94.5%/90.4%, 53.0%/60.0%, 86.2%/84.4%, and 47.3%/50.8%, respectively. Accuracy of tumor detection varied widely per anatomic site. Statistically significant interobserver differences were noted, specifically for tumor size of 1-5 cm (P = 0.007) and localization on mesentery and small bowel (kappa = 0.30, P = 0.04). CONCLUSIONS: In colorectal cancer, CT detection of peritoneal carcinomatosis is moderate and of individual peritoneal tumor deposits poor. Interobserver differences are statistically significant. Therefore, preoperative CT seems not to be a reliable tool for detection of presence, size, and location of peritoneal tumor implants in view of treatment planning in patients with colorectal cancer. PMID- 15112248 TI - Efficacy of sentinel lymph node biopsy in male breast cancer. AB - BACKGROUND: Sentinel lymph node biopsy (SLNB) is rapidly becoming the standard of care in the treatment of women with early stage breast cancer. Male breast cancer although relatively rare, has typically been treated with mastectomy and axillary lymph node dissection (ALND). Men who develop breast carcinoma have the same risk as their female counterparts of developing the morbidities associated with axillary dissection. SLNB has been championed as a procedure aimed at preventing those morbidities. We recently have evaluated the role of SLNB in the treatment of men with early stage breast cancer. METHODS: Among the 18 men treated at the University of Michigan Medical Center for breast cancer from May 1998 to November 2002, 6 were treated with SLNB. RESULTS: The mean tumor size was 1.6 cm. The mean patient age was 59.8 years. All of the patients had one or more sentinel lymph nodes identified. Two of the six did not have confirmatory axillary dissection. Three of the six had positive sentinel lymph nodes (50%). Only one of the three patients with a positive sentinel node had more nodes positive. One of the six patients had a positive node on frozen section and underwent immediate complete axillary dissection. This patient had no additional positive nodes. No patients in our series had immunohistochemical studies of the lymph nodes. CONCLUSIONS: Men with early stage breast carcinoma may be offered the management option of SLNB since in the hands of experienced surgeons it has a success rate apparently equal to that in their female counterparts. PMID- 15112249 TI - Primary malignant tumors of the iliopsoas compartment. AB - BACKGROUND AND OBJECTIVES: Primary malignant tumors of the iliopsoas compartment are rare entities that have been infrequently reported. We present our experience of iliopsoas malignancy to better characterize and define the natural history of this condition. METHODS: Primary iliopsoas tumors occurring from January 1990 were identified from the Royal Marsden Hospital's (RMH) Sarcoma Unit prospective database. RESULTS: Nineteen malignant tumors (11 limited to the psoas muscle only, 1 involving only the iliacus muscle and 7 involving both the iliacus and psoas) of the iliopsoas compartment were evaluated and treated at the RMH during this period. Leiomyosarcoma (n = 5) and liposarcoma (n = 3) were the most frequent histologic types. There were 3 G1, 5 G2, 10 G3 tumors and grade could not be assessed in 1 patient. There were 17 T2 tumors and size was not available in 2 patients. Surgery was done in 14 patients (negative microscopic margins-7, positive microscopic margins-3, and positive gross margins-3 and the margin of excision was not known in 1 patient). Five of 14 surgically treated patients had local recurrence and 6 of 19 patients developed metastases. Ten patients died of their disease at a median follow-up of 12 months. The estimated 2 and 5 year survival rates were 44 and 23%, respectively. CONCLUSIONS: Tumors in this location have a poor prognosis due to the lack of early diagnosis, large size at presentation, multiple attachments of the psoas muscle, and being relatively surgically inaccessible. PMID- 15112250 TI - Prognostic value of von Willebrand factor, CD34, CD31, and vascular endothelial growth factor expression in women with uterine leiomyosarcomas. AB - BACKGROUND AND OBJECTIVES: To compare uterine leiomyosarcomas (LMS) and leiomyomas (LM) with normal myometrium in terms of microvessel density (MVD), and to correlate this parameter with vascular endothelial growth factor (VEGF) expression and clinical/pathological parameters. METHODS: An immunohistochemical technique, using antibodies against von Willebrand factor (FvW), CD34, CD31, and VEGF, was applied to formalin-fixed paraffin-embedded samples of 32 normal myometria, 32 uterine LM, and 12 LMS. MVD was calculated by a digital image analyzer. RESULTS: Using anti-FvW, mean +/- SD MVD in myometrium, LM, and LMS was 107.0 +/- 53.6, 66.2 +/- 55.4, and 64.4 +/- 44.2, respectively (P = 0.001). MVD was lower in LMS (P = 0.021) and in LM (P = 0.0004) than in normal myometrium. Using anti-CD34, mean +/- SD MVD in myometrium, LM, and LMS was 187.6 +/- 91.2, 106.1 +/- 55.5, and 114.2 +/- 98.8, respectively (P = 0.001). MVD was lower in LMS (P = 0.012) and LM (P = 0.0004) than in normal myometrium. No such differences were found using anti-CD31 and anti-VEGF. No correlation was found between MVD and VEGF expression. In women with uterine LMS, low MVD (assessed with anti-FvW) correlated with recurrence (P = 0.04) and poor overall survival (P = 0.03). CONCLUSIONS: Uterine smooth muscle tumors exhibit a lower MVD than normal myometrium, as assessed using anti-FvW or anti-CD34 antibodies. A reduced MVD, as assessed by FvW staining, has prognostic value in uterine LMS. PMID- 15112251 TI - Nutritional status and cytokine-related protein breakdown in elderly patients with gastrointestinal malignancies. AB - BACKGROUND: Age-related factors affecting cytokine-related whole-body protein breakdown and their relation to clinical outcomes in cancer patients were investigated. METHODS: For assessment of protein-calorie malnutrition and protein breakdown, the creatinine height index (CHI) and daily urinary excretion of 3 methylhistidine (3-MH) were measured in 70 patients with gastrointestinal malignancies. Perioperative cytokine profile was evaluated to assess its relation to perioperative protein catabolism. RESULTS: In elderly patients, daily 3-MH excretion during the stable preoperative period decreased with the increase of tumor interleukin (IL)-6 production, suggestive of the activation of a metabolic compensation mechanism. However, these patients showed significant increases in postoperative 3-MH excretion in accord with perioperative systemic IL-6 response, and this deterioration of the compensating mechanism seemed to be associated with poor clinical outcome. An increase in 3-MH excretion under surgical stress was positively correlated with postoperative consumption of IL-6 soluble receptor (sR) in elderly patients with nutritional depletion. CONCLUSIONS: In elderly cancer patients with protein-calorie malnutrition, metabolic compliance against intrinsic IL-6 may be compensated for in the preoperative stable period, but deteriorate from surgical insults. This mechanism might involve increased affinity of IL-6 with IL-6sR under surgical stress. PMID- 15112252 TI - Telomerase activity and expression of telomerase genes in squamous dysplasia and squamous cell carcinoma of the esophagus. AB - BACKGROUND: Telomerase maintains telomere length and is considered to be necessary for the indefinite proliferation of human cells. Activation of telomerase plays a key role in the malignant transformation process. The aim of this study was to study the regulation of telomerase, and to explore the possibility of telomerase as a biomarker in squamous carcinogenesis of the esophagus. METHODS: Twenty-nine esophageal squamous cell carcinomas (ESCC) and its corresponding adjacent normal tissues, and 47 epithelial squamous dysplasia tissues were analyzed by the reverse transcriptase-polymerase chain reaction (RT PCR) technique for the mRNA expression of three major telomerase subunits: human telomerase RNA (hTR), telomerase protein component 1 (TP1), and human telomerase reverse transcriptase (hTERT) and by telomeric repeat amplification protocol assay (TRAP) for telomerase activity. RESULTS: For the expression of hTR and TP1 mRNA, there were no significant differences among ESCC, dysplasia and normal tissues (P > 0.05). In contrast, hTERT mRNA expression was detected in 28 of 29 ESCC (96.6%), in 23 of 47 dysplasia (48.9%), and only in two of 29 normal tissues (7.5%). Telomerase activity was positive in 25 of 29 ESCC (86.2%), in 21 of 47 (44.7%) epithelial dysplasia tissues, and in none of normal tissue. All together, 95 of 105 cases (90.48%) were concordant for both results, i.e., telomerase activity positive and hTERT positive or telomerase activity negative and hTERT negative tissues, and telomerase activity correlated with hTERT mRNA expression (P < 0.001). CONCLUSIONS: Higher telomerase activity and hTERT mRNA expression were shown during an early stage in the esophageal carcinogenesis. Activation of telomerase activity was strongly correlated with hTERT mRNA expression, suggesting hTERT is a major regulator of telomerase activity, and telomerase activation may play a critical role in esophageal carcinogenesis. Therefore, telomerase, especially hTERT can be used as a potential molecular biomarker of ESCC. PMID- 15112253 TI - A novel technique of raising a pectoralis major myocutaneous flap through the skin paddle incision alone. AB - A good reconstruction should not only be functionally and aesthetically sound at the recipient site but also cause least possible cosmetic aberration of the donor site. The pectoralis major myocutaneous (PMMC) flap continues to be one of the most commonly used flap for head and neck reconstruction in this part of the world. Conventionally, once the skin paddle over the pectoralis major muscle is marked, a line is drawn joining the outer edge of the skin flap extending to the apex of the anterior axillary skin fold or midclavicular point to expose the underlying pectoralis major muscle and harvest the flap. We intend to suggest a novel technique, in which the pectoralis major muscle is exposed by raising the skin around the skin paddle incision alone without making any further extension. PMID- 15112254 TI - Primary retroperitoneal teratomas: a review of the literature. AB - Teratomas are uncommon neoplasms comprised of mixed dermal elements derived from the three germ cell layers. Historically, teratomas were attributed to demons, sexual misconduct, and abnormal fertilization. They attract attention because of their bizarre histology and gross appearance. While the majority of teratomas present congenitally in the sacrococcygeal region, within the ovaries of adolescent females and within the testes of young men, they have been identified throughout the body. Extragonadal teratomas tend to occur in midline structures as the anterior mediastinum, retroperitoneum, sacrococcygeal region, and pineal gland. Retroperitoneal teratomas represent only 1-11% of primary retroperitoneal tumors. Incidence is bimodal with peaks in the first 6 months of life and in early adulthood. Due to their location, they are usually identified only after they have grown to huge proportions. There is a 25% chance of malignancy. Surgical resection remains the mainstay of therapy and is required for definitive diagnosis. This article reviews the literature on the histopathology, classification, genetic abnormalities, and theories of origin of teratomas as well as the presentation, diagnosis, and management of retroperitoneal teratomas. PMID- 15112255 TI - The challenge of discrepancies in values among physicians, patients, and family members. PMID- 15112256 TI - Diagnostic strategies for unknown primary cancer. AB - Unknown primary cancer (UPC) is defined by the presence of metastatic disease for which a primary site is undetectable on presentation. Computed tomography scan of the body was performed routinely in search of the primary cancer and invasive procedures were pursued in selective cases. Magnetic resonance imaging of the breast enables identification of an occult breast primary tumor in < or = 75% of women who present with adenocarcinoma in the axillary lymph nodes and can influence surgical management. Positron emission tomography scan also can be used in the diagnosis of UPCs, but its value is controversial. Cytokeratins 7 and 20 and thyroid transcription factor are some of the histochemical markers used in most patients who present with metastatic adenocarcinoma. Some of the newly discovered immunohistochemical markers further assist in narrowing the differential diagnosis. The role of molecular profiling to make the diagnosis, establish the prognosis, and assess the response to treatment in UPCs is evolving. The authors discuss the role of histochemical markers in the diagnosis of UPC and the most recent data regarding the use of imaging and invasive diagnostic modalities and gene expression profiles. PMID- 15112257 TI - Management of T1-T2 glottic carcinomas. AB - T1-T2 glottic carcinomas may be treated with conservative surgery or radiotherapy. The goals of treatment are cure and laryngeal voice preservation. The aim of the current study was to review the pertinent literature and discuss the optimal management of early-stage laryngeal carcinoma. Literature review indicated that the local control, laryngeal preservation, and survival rates of patients were similar after transoral laser resection, open partial laryngectomy, and radiotherapy. Voice quality depended on the extent of resection for patients undergoing surgery; results for patients undergoing laser resection for limited lesions were comparable to the corresponding results for patients receiving radiotherapy, whereas open partial laryngectomy yielded poorer results. Costs were similar for laser resection and radiotherapy, but open partial laryngectomy was more expensive. Patients with well defined lesions suitable for transoral laser excision with a good functional outcome were treated with either laser or radiotherapy. The remaining patients were optimally treated with radiotherapy. Open partial laryngectomy was reserved for patients with locally recurrent tumors. PMID- 15112258 TI - Gemcitabine-related radiation recall preferentially involves internal tissue and organs. AB - Radiation recall refers to inflammatory reactions triggered by cytotoxic agents and develops in previously irradiated areas. Most reactions develop cutaneously. The most common chemotherapeutic agents implicated are anthracyclines and taxanes. Gemcitabine, a nucleotide analog, recently was implicated in several cases. The authors performed a literature search using PubMed and the search terms "gemcitabine" and "radiation recall" to find prior cases of radiation recall attributed to gemcitabine. These cases were compared with those attributed to anthracyclines and taxanes. The literature search found 12 cases of radiation recall caused by gemcitabine. The authors also determined that their case of myositis developing in the rectus abdominus muscle of a patient with pancreatic adenocarcinoma was the manifestation of radiation recall, thereby bringing the number of patients who developed radiation recall to gemcitabine and were discussed in the current study to 13. Approximately 70% of the cases manifested as inflammation of internal organs or tissues and 30% manifested as a dermatitis or mucositis. This finding differs from other common agents, in which 63% of the radiation recall events are reported to manifest as a dermatitis. Compared with anthracyclines and taxanes, the interval from the completion of radiation therapy to the initiation of chemotherapy is less for gemcitabine (median time of 56 days for gemcitabine, compared with 218 days for the taxanes and 646 days for doxorubicin). The majority of radiation recall reactions attributed to gemcitabine are reported to affect internal tissue or organs. In contrast, other common agents for the most part trigger cutaneous inflammation. The development of internal tissue inflammation is reportedly correlated with a shorter interval from the time of completion of radiation therapy to the initiation of chemotherapy. PMID- 15112259 TI - Acceptance of tamoxifen chemoprevention by physicians and women at risk. AB - BACKGROUND: In the National Surgical Adjuvant Breast and Bowel Project (NSABP) P 1 trial, tamoxifen was shown to reduce breast carcinoma risk by 49% in high-risk women. The purpose of the current study was to identify factors associated with being offered, and accepting, tamoxifen chemoprevention. METHODS: The records of 219 women who sought risk evaluation after the publication of the NSABP P-1 trial between September 1998 and October 2002 were reviewed. Risk was calculated using the model of either Gail et al. or Claus et al. The impact of individual risk factors on the offering and acceptance of tamoxifen was compared using the Fisher exact test and logistic regression analysis. RESULTS: Tamoxifen was offered to 137 women (63%) in the current study. The magnitude of Gail risk, age, menopausal status, hysterectomy, and history of lobular carcinoma in situ (LCIS) or atypical hyperplasia (AH) were all found to be significant predictors of a patient being offered tamoxifen. On multivariate analysis, only a history of AH or LCIS and hysterectomy were found to be significant, with odds ratios of 20.3 and 3.4, respectively. Fifty-seven of the women who were offered tamoxifen (42%) took the drug. Only a history of LCIS or AH and older age were found to be predictive of tamoxifen acceptance. CONCLUSIONS: In the current study, risk due to AH or LCIS was found to be the main predictor of being offered and accepting tamoxifen chemoprevention. PMID- 15112260 TI - The natural history of breast carcinoma in the elderly: implications for screening and treatment. AB - BACKGROUND: The authors evaluated the two indicators of metastatic proclivity (namely, virulence [V; the rate of appearance of distant metastases] and metastagenicity [M; the ultimate likelihood of developing distant metastases]) of breast carcinoma in elderly women. The authors then compared these characteristics with the corresponding characteristics in a cohort of younger women to determine whether breast carcinoma was more indolent in women age > 70 years, as is commonly believed in the medical community. METHODS: The authors examined 2136 women who underwent mastectomy without adjuvant systemic therapy at The University of Chicago Hospitals (Chicago, IL) between 1927 and 1987. The median follow-up period was 12.3 years. Distant disease-free survival (DDFS) was determined for women who did not receive systemic therapy. V and M were obtained from log-linear plots of DDFS. RESULTS: No significant difference in tumor size at presentation was observed among women age < 40 years, women ages 40-70 years, and women age > 70 years (P = 0.86), whereas significantly fewer women age > 70 years presented with positive lymph nodes compared with younger women (P = 0.05). In women with negative lymph node status, there was a higher DDFS rate among patients ages 40-70 years (81% at 10 years) compared with patients age > 70 years (65% at 10 years; P = 0.018). There was no significant age-related difference among women with lymph node-positive disease (P = 0.2). For example, the 10-year DDFS rate for women ages 40-70 years was 33%, compared with 38% for women age > 70 years. Among those with lymph node-negative disease, V was 3% per year for women ages 40-70 years as well as women age > 70 years. Among women with lymph node-negative disease, M was 0.20 for patients ages 40-70 years and 0.35 for patients age > 70 years. In women with positive lymph node status, both V (11% per year vs. 10% per year) and M (0.70 vs. 0.65) were similar in both age groups. CONCLUSIONS: Fewer women age > 70 years had lymph node involvement at presentation. However, when this finding was taken into account, the authors found no evidence that breast carcinoma was more indolent in women age > 70 years. These results support the use of similar diagnostic and therapeutic efforts for elderly women and younger women, with modification for elderly women based only on comorbidity. PMID- 15112261 TI - Pharmacoproteomic analysis of prechemotherapy and postchemotherapy plasma samples from patients receiving neoadjuvant or adjuvant chemotherapy for breast carcinoma. AB - BACKGROUND: In this study, proteomic changes were examined in response to paclitaxel chemotherapy or 5-fluorouracil, doxorubicin, and cyclophosphamide (FAC) chemotherapy in plasma from patients with Stage I-III breast carcinoma. The authors also compared the plasma profiles of patients with cancer with the plasma profiles of healthy women to identify breast carcinoma-associated protein markers. METHODS: Sixty-nine patients and 15 healthy volunteers participated in the study. Plasma was sampled on Day 0 before chemotherapy and on Day 3 posttreatment in the 69 patients or 3 days apart in the 15 healthy women. Twenty nine patients received preoperative chemotherapy, and 40 received postoperative chemotherapy. Surface-enhanced laser desorption/ionization mass spectrometry was used to generate protein mass profiles. RESULTS: Few changes were observed in plasma during treatment. Only 1 protein peak was identified (mass/charge ratio [m/z], 2790) that was induced by paclitaxel and, to a lesser extent, by FAC chemotherapy. This proteomic response was detectable in 80% of patients who were treated preoperatively but also was present with lesser intensity in approximately 40% of patients treated postoperatively. There was no clear correlation between induction of m/z 2790 during a single course of treatment and final tumor response to preoperative chemotherapy. Five other peaks also were identified that discriminated between plasma from patients with breast carcinoma and plasma from normal women. These same peaks also were detectable in a subset of patients who already had undergone surgery to remove their tumors. CONCLUSIONS: A single chemotherapy-inducible SELDI-MS peak and five other peaks that distinguished plasma obtained from patients with breast carcinoma from plasma obtained from normal, healthy women were identified. The (as yet unsequenced) proteins represented by these peaks are candidate markers of micrometastatic disease after surgery. PMID- 15112262 TI - Effect of margins on ipsilateral breast tumor recurrence after breast conservation therapy for lymph node-negative breast carcinoma. AB - BACKGROUND: Breast conservative surgery (CS) with radiotherapy (RT) is the most commonly used treatment for early-stage breast carcinoma. However, there is controversy regarding the importance of the pathologic margin status on the risk of ipsilateral breast tumor recurrence (IBTR). The current study evaluated the effect of the pathologic margin status on IBTR rates in a cohort of women with lymph node-negative breast carcinoma treated with CS and RT. METHODS: Between August 1980 and December 1994, 452 women with pathologically lymph node-negative breast carcinoma were treated with CS and RT at Westmead Hospital (Westmead, Australia). Central pathology review was performed for all women. The final margins were negative for 352 women (77.9%), positive (invasive and/or in situ) for 42 women (9.3%), and indeterminate for 58 women (12.8%). Information regarding an extensive intraductal component (EIC), lymphovascular invasion, pathologic tumor size, histologic grade, and nuclear grade was available for most women. After macroscopic total excision of the tumor, all women received whole breast irradiation (usually 45-50.4 grays [Gy]) and the majority of women also received a local tumor bed boost (range, 8-30 Gy). RESULTS: After a median follow up of 80 months, 34 women (7.5%) developed an IBTR. The crude 5-year rates of IBTR for women with negative margins, positive margins, and indeterminate margins were 3.1%, 11.9%, and 6.9%, respectively. For women with negative margins, the 5 year and 10-year actuarial rates of freedom from IBTR were 96% and 92%, respectively, compared with 88% and 75%, respectively, for women with positive margins (P = 0.003). Univariate analysis demonstrated that the only factors associated with a significantly higher risk of IBTR were age at diagnosis (P < 0.050) and margin status (P = 0.005). Multivariate analysis showed that both age and margin status were independent predictors of IBTR. None of 24 patients with an EIC and negative margins were found to have developed an IBTR. CONCLUSIONS: The results of the current study were comparable to other published reports and supported the association of higher IBTR rates with positive or indeterminate margins compared with negative, pathologic margins. Furthermore, young age (age < 35 years at diagnosis) was associated with the highest risk of IBTR regardless of margin status. PMID- 15112263 TI - Initially metastatic breast carcinoma has a distinct disease pattern but an equivalent outcome compared with recurrent metastatic breast carcinoma. AB - BACKGROUND: To date, the clinical features and outcomes of patients with initially metastatic breast carcinoma (IMBC) have not been compared with the corresponding characteristics in patients with recurrent metastatic breast carcinoma (RBC). This issue may be particularly relevant to clinical research, as it may shed light on a potential bias with respect to the selection of patients for clinical trials. METHODS: A retrospective analysis of the medical records of 1350 patients with breast carcinoma was performed. Outcome variables included overall survival, response rate, and progression-free survival. RESULTS: One hundred nineteen of 370 patients with metastatic breast carcinoma had IMBC, whereas the remaining 251 had RBC. The median follow-up duration was 39.4 months, and the median overall survival duration was 24 months. With regard to clinical characteristics, patients with IMBC were older than patients with RBC (61.7 years vs. 58.1 years; P < 0.001) and had a higher incidence of lobular carcinoma (15.9% vs. 7.7%; P = 0.018), a greater proportion of T3-4 tumors (58.8% vs. 27.9%; P < 0.001), a higher incidence of bone as the dominant metastatic site (41.2% vs. 21.5%; P < 0.001), a lower incidence of soft tissue as the dominant metastatic site (10.1% vs. 26.7%; P < 0.001), and a similar incidence of the viscera as the dominant metastatic site (48.7% vs. 51.8%; P = 0.78). Median overall survival duration was similar for patients with IMBC (25.1 months) and patients with RBC (23.3 months; P = 0.81). Statistical analyses also revealed nonsignificant differences between patients with IMBC and patients with RBC in terms of response rate (40.7% vs. 35.2%, respectively; P = 0.35) and median progression-free survival duration (10.2 months vs. 9.0 months, respectively; P = 0.58). CONCLUSIONS: Although patients with IMBC and patients with RBC exhibit distinct histologic and clinical characteristics, similar treatment efficacy results and survival outcomes are observed in these two groups. PMID- 15112264 TI - Primary care provider perceptions of barriers to and facilitators of colorectal cancer screening in a managed care setting. AB - BACKGROUND: Colorectal cancer (CRC) screening tests (e.g., fecal occult blood testing [FOBT], flexible sigmoidoscopy [FS], etc.) are underused. Primary care providers (PCPs) play a critical role in screening, but barriers to and facilitators of screening as perceived by PCPs in managed care settings are poorly understood. The objectives of the current study were to describe current CRC screening practices and to explore determinants of test use by PCPs in a managed care setting. METHODS: In 2000, a self-administered survey was mailed to a stratified, random sample of 1340 PCPs in a large, network model health maintenance organization in California. RESULTS: The survey response rate was 67%. PCPs indicated that 79% of their standard-risk patients were screened for CRC. PCP-reported median rates of recommendation for the use of specific screening tests were 90% for FOBT and 70% for FS. In logistic regression models, perceived barriers to the use of FOBT and FS included patient characteristics (e.g., education) and PCP-related barriers (e.g., failure to recall that patients were due for testing). Perceived facilitators of the use of FOBT and FS included interventions targeting certain aspects of the health care system (e.g., reimbursement) and interventions targeting certain aspects of the tests themselves (e.g., provision of evidence of a test's effectiveness). Assignment of high priority to screening, integrated medical group (as opposed to independent practice association) affiliation, and the proportion of patients receiving routine health maintenance examinations were positively associated with reported test use. CONCLUSIONS: CRC screening tests appear to be underused in the managed care setting examined in the current study. The perceived barriers and facilitators that were identified can be used to guide interventions aimed at increasing recommendations for, as well as actual performance of, CRC screening. PMID- 15112265 TI - Methyl-CpG-binding domain 2: a protective role in bladder carcinoma. AB - BACKGROUND: MBD2, a methyl-CpG-binding domain 2 protein, has attracted much attention because of its role in epigenetic regulation of gene expression. In addition to transcriptional repression, MBD2 has also been shown to catalyze demethylation by directly removing methyl groups from 5-methylcytosine residues in DNA. Although the demethylase activity of MBD2 remains controversial, reduction of MBD2 messenger RNA expression has been observed in various tumor tissue types. In the current case-control study, the authors investigated the association between MBD2 expression and bladder carcinoma risk. METHODS: RNA was isolated from the peripheral blood lymphocytes of 98 bladder carcinoma case patients and 135 frequency-matched control patients. MBD2 expression was measured by real-time quantitative reverse transcription-polymerase chain reaction assays. RESULTS: Overall, there was a significantly reduced risk associated with high levels of MBD2 expression (odds ratio [OR], 0.43; 95% confidence interval [CI], 0.21-0.90). This relation was maintained when the data were categorized according to quartile distribution for MBD2 expression (P for trend < 0.05). It is noteworthy that the protective effects were more apparent in women (OR, 0.25; 95% CI, 0.06-1.02) compared with men (OR, 0.58; 95%; CI, 0.24-1.42), in older individuals (OR, 0.12; 95% CI, 0.03-0.45) compared with younger individuals (OR, 1.16; 95% CI, 0.40-3.33), and in heavier smokers (OR, 0.40; 95% CI, 0.18-0.93) compared with lighter smokers (OR, 0.71; 95% CI, 0.18-2.86). CONCLUSIONS: Although the underlying molecular mechanisms remain unclear, the data obtained in the current study represent the first evidence demonstrating a protective role against bladder carcinoma risk for MBD2. MBD2 expression may prevent age-related, gender-related, and smoking-induced hypermethylation, which are predisposing factors for tumor development. PMID- 15112266 TI - p53 and p21 Expression levels predict organ preservation and survival in invasive bladder carcinoma treated with a combined-modality approach. AB - BACKGROUND: The purpose of the current study was to evaluate the expression levels of p53, p21 and pRB as predictors of for long-term organ preservation and survival in patients with bladder carcinoma who were treated with bladder-sparing intent using a combined-modality approach. METHODS: Tumor samples from 82 consecutive patients with localized invasive bladder carcinoma treated on 3 different bladder-sparing studies were examined for p53, p21, and pRB expression by immunohistochemical methods. Treatment consisted of transurethral resection, platinum-based neoadjuvant chemotherapy, and, according to response, either radiotherapy or radical cystectomy. The median follow-up duration was 55 months. RESULTS: Positive immunoreactivity for p53, p21, and pRB was observed in 47%, 52%, and 67% of patients, respectively. Positive p53 immunoreactivity and positive p21 immunoreactivity were independent predictors of decreased survival with bladder preservation (P = 0.02 and P = 0.02, respectively) and disease-free survival (DFS; P = 0.005 and P = 0.009, respectively) in a multivariate analysis adjusting for clinical stage, ureteral obstruction, and age. Regarding overall survival (OS), p53 overexpression was associated with poor outcome (P = 0.03), whereas the association of poor outcome with p21 expression did not reach statistical significance (P = 0.07). No association between pRB immunoreactivity and outcome was found. When the combined expression of p53 and p21 was assessed, the positive expression of both markers was a strong and unfavorable prognostic factor for survival with bladder preservation (P = 0.006), DFS (P = 0.003), and OS (P = 0.02). CONCLUSIONS: Expression levels of p53 and p21, especially when simultaneously assessed, exhibit independent predictive value for long-term bladder preservation and survival in patients with bladder carcinoma treated with combined-modality therapy. These determinations could be useful in the selection of candidates for bladder-preserving treatment. PMID- 15112267 TI - High-dose calcitriol, zoledronate, and dexamethasone for the treatment of progressive prostate carcinoma. AB - BACKGROUND: Preclinical and clinical data have suggested that high-dose calcitriol (1,25-dihydroxycholecalciferol) has activity against prostate carcinoma. Pulse-dosed calcitriol and dexamethasone may maximize tolerability and efficacy. The authors examined the toxicity of pulse-dosed calcitriol with zoledronate and with the addition of dexamethasone at the time of disease progression. METHODS: Patients with progressive prostate carcinoma were eligible for the current study. In cohorts of 3-6 patients, calcitriol was administered for 3 consecutive days per week, starting at a dose of 4 microg per day. Doses were escalated to 30 microg per day. Intravenous zoledronate (4 mg) was administered monthly. Dexamethasone could be added to the regimen at disease progression. Toxicities, markers of bone turnover, plasma calcitriol levels, and clinical outcomes were recorded. RESULTS: Thirty-one patients were treated in cohorts that were defined by the calcitriol dose administered (4, 6, 8, 10, 14, 20, 24, or 30 microg). Seven patients received dexamethasone. Three patients had their doses reduced due to calcium-related laboratory findings. Patients tolerated therapy well, even in the 30 microg cohort; therefore, a maximum tolerated dose was not defined. Peak plasma levels observed in the 24 microg and 30 microg cohorts ranged from 391 to 968 pg/mL. Minimal antitumor effects were observed. CONCLUSIONS: Calcitriol was well tolerated at doses up to and including 30 microg 3 times per week in combination with intravenous zoledronate 4 mg monthly, with or without dexamethasone, in patients with progressive prostate carcinoma. Peak plasma levels in the 24 microg and 30 microg cohorts were greater than the levels associated with antitumor effects preclinically. Due to the cumbersome dosing schedule and the lack of significant activity observed, Phase II trials of this regimen are not planned. PMID- 15112268 TI - Diagnostic accuracy of fine-needle aspiration cytology and frozen section in primary parotid carcinoma. AB - BACKGROUND: The low incidence and histologic heterogeneity of primary parotid carcinomas makes it difficult to evaluate the value of preoperative fine-needle aspiration cytology (FNAC) and intraoperative frozen section (FS) analysis. In the current study, the authors reviewed a single institution's experience regarding the preoperative and intraoperative diagnostic value of FNAC and FS in primary salivary gland carcinomas. METHODS: Between January 1990 and December 2002, 108 primary parotid carcinomas were resected at the Department of Otorhinolaryngology, Head and Neck Surgery at the University of Berne, Inselspital (Berne, Switzerland). Included in the study were a total of 101 carcinomas with preoperative FNAC results in 88 tumors and/or intraoperative FS results in 45 tumors. In a retrospective study, the results of FNAC and FS were analyzed and compared with the corresponding histopathologic diagnoses. RESULTS: The cytologic findings were true-positive for malignancy in 63 tumors (72%), false-negative in 22 tumors (25%), and nondiagnostic in 3 tumors (3%). The tumor grading was correct in 29 of 63 tumors (46%), and the exact tumor typing was correct in 27 of 63 (43%) true-positive tumors. The FS findings were true positive for malignancy in 43 of 45 tumors (96%), the tumor grading was correct in 35 of 45 tumors (78%), and the tumor typing was correct in 32 of 45 tumors (71%). Overall, at the time of surgery, of the 101 parotid carcinomas, the tumor was known to be malignant in 83 tumors (82%), and the correct grade and the exact tumor type were known in 55 tumors (54%) and 48 tumors (48%), respectively. CONCLUSIONS: FNAC recognized malignancy in 72% of tumors, but it could not be relied upon to provide an accurate tumor grading or typing. Therefore, FNAC alone is not prone to determine the surgical management of primary parotid carcinomas. The current analysis showed the statistically significant superiority of FS compared with FNAC regarding the diagnosis of malignancy, tumor grading, and tumor typing in primary parotid carcinomas. PMID- 15112269 TI - Soluble vascular endothelial growth factor receptor 1, and not receptor 2, is an independent prognostic factor in acute myeloid leukemia and myelodysplastic syndromes. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) and its receptors (VEGFRs) are major regulators of angiogenesis, which plays a key role in the growth and dissemination of solid tumors and hematologic neoplasms. METHODS: The authors measured the plasma concentrations of soluble VEGFR1 (sVEGFR1) and sVEGFR2 in 133 patients with acute myeloid leukemia (AML) and in 80 patients with myelodysplastic syndromes (MDS) at the time of initial presentation and compared clinical behaviors. RESULTS: A reverse correlation was observed between plasma sVEGFR1 levels and the rate of complete remission (CR) in patients with AML, but not in patients with MDS. In contrast, increased plasma levels of sVEGFR2 were correlated with a lower CR rate in patients with MDS, but not in patients with AML. Cox regression model analysis demonstrated that plasma levels of sVEGFR1, but not sVEGFR2, were independent prognostic factors in both patients with AML and patients with MDS. CONCLUSIONS: The findings suggest that different mechanisms are involved in the pathophysiology of AML and MDS. The concentration of sVEGFR1 and sVEGFR2 in plasma should be considered a significant factor in guiding antiangiogenic therapy for AML and MDS. They may play a role in the pharmacodynamics of therapeutic agents that are supposed to bind directly to these receptors. PMID- 15112270 TI - Adoptive immunotherapy with allogeneic Epstein-Barr virus (EBV)-specific cytotoxic T-lymphocytes for recurrent, EBV-positive Hodgkin disease. AB - BACKGROUND: It has been shown that adoptive immunotherapy with Epstein-Barr virus (EBV)-specific cytotoxic T-lymphocytes (CTL) is effective for the treatment of EBV-induced lymphoproliferative disease in stem cell transplantation recipients and organ transplantation recipients. The role of EBV CTL in other tumors for which this virus has been implicated in pathogenesis, such as EBV-positive Hodgkin disease (HD), has not been demonstrated clearly. METHODS: To investigate the antitumor effects and toxicity of allogeneic EBV CTL in EBV-positive HD, the authors initiated a pilot trial in which EBV CTL were cultured from allogeneic, partially human leukocyte antigen-matched donors and were infused into patients who had therapy-refractory disease. The first cohort of 3 patients (Cohort I) received 3 separate infusions of EBV CTL (5.0 x 10(6) EBV CTL/kg per dose), and the second cohort (Cohort II) received 30 mg/m(2) per day of fludarabine for 3 days followed by a single CTL infusion (1.5 x 10(7) EBV CTL/kg). RESULTS: All three patients in Cohort I had decreases in measurable disease after EBV CTL infusions, and one of those patients was without evidence of disease 22 months after infusion. Two of 3 patients in Cohort II had decreases in measurable disease, although it was not determined whether those decreases were related to fludarabine or to CTL, and 1 patient in Cohort II had 7 months without disease progression. Unlike the patients in Cohort I, fludarabine recipients did not have increases in antidonor CTL responses. Donor cells could not be detected in any of the CTL recipients. CONCLUSIONS: Adoptive immunotherapy with allogeneic EBV CTL was safe for patients with recurrent, refractory, EBV-positive HD; and clinical responses may be observed without the establishment of detectable donor lymphoid chimerism. PMID- 15112271 TI - Familial aggregation of Hodgkin lymphoma and related tumors. AB - BACKGROUND: The importance of genetic factors in the etiology of Hodgkin lymphoma (HL) has been suggested by family and population studies. However, the spectrum of malignancies associated with common genetic etiology and the effects of gender and age on familial risk have not been established. METHODS: Diagnoses of lymphoproliferative malignancies were compared in 15,799 first-degree relatives of 5047 patients with HL versus 32,117 first-degree relatives of 10,078 control probands from Sweden and in 7185 first-degree relatives of 2429 patients with HL versus 27,434 first-degree relatives of 8,495 control probands from Denmark using marginal survival models. RESULTS: The risk of HL in relatives of patients with HL was increased significantly in both populations, with relative risks of 3.47 (95% confidence interval [95% CI], 1.77-6.80) in Sweden and 2.55 (95% CI, 1.01 6.45) in Denmark and a pooled estimate of 3.11 (95%CI, 1.82-5.29). In Sweden, risks for relatives of patients also were increased significantly for chronic lymphocytic leukemia and non-Hodgkin lymphoma (in males). Relative risks were higher in males compared with females and in siblings of patients compared with parents and offspring of patients. Relatives of patients with earlier-onset disease were at higher risk for HL. CONCLUSIONS: HL has an important familial component, which is stronger in families of affected individuals age < 40 years, in males, and in siblings, and it is shared with some (but not other) lymphoproliferative malignancies. The cumulative lifetime risks are very small, however, for the development of HL de novo or in first-degree relatives of affected patients. PMID- 15112272 TI - Skip metastasis in nonsmall cell lung carcinoma: predictive markers and isolated tumor cells in N1 lymph nodes. AB - BACKGROUND: Skip metastasis to mediastinal lymph nodes is a prognostic factor for patients with nonsmall cell lung carcinoma (NSCLC). Little is known about the biologic behavior of tumors with noncontinuous spread to the mediastinal lymph nodes. In patients with pN2 skip metastases, micrometastases to N1 lymph nodes, which only mimic skip metastases, have not been investigated. METHODS: In a retrospective study, the authors analyzed the primary tumor specimens from 45 patients with pN2 NSCLC (18 patients had squamous cell carcinomas, 23 had adenocarcinomas, and 4 had large cell carcinomas). They immunohistochemically evaluated the expression of p21, p53, MUC-1, Bcl-2, c-ErbB-2, and E-cadherin. Survival rates and biomarker expression levels were compared between patients with pN2 disease and infiltration of N1 lymph nodes (without skip metastasis [n = 28]) and patients with pN2 disease without N1 infiltration (with skip metastasis [n = 17]). To evaluate micrometastasis in the pN1 lymph nodes of 17 patients with skip metastases, lymph nodes were stained using the anticytokeratin antibody, AE1/AE3. RESULTS: The 5-year survival rate of patients with skip metastases was 41%, compared with 14% for patients without skip metastases (P = 0.019). In a multivariate analysis, the incidence of skip metastases did not vary significantly according to gender, age, histology, pT status, or cM status. Three skip-positive patients (17.6%) had micrometastatic tumor involvement of pN1 lymph nodes. After adding these patients to the group of patients without skip metastases, there was still a significant difference in survival between the two groups. p53, MUC-1, c-ErbB-2, and E-cadherin expression levels in primary tumor specimens were not significantly different in patients with continuous metastasis and patients with skip metastases. Patients with skip metastases expressed lower levels of p21 (P = 0.026), whereas Bcl-2 expression levels were considerably higher (P = 0.019) compared with the corresponding levels in patients without skip metastases. CONCLUSIONS: Patients with NSCLC and pN2 skip metastases have a more favorable prognosis than do patients with pN2 disease without skip metastases. Tumor specimens from these patients exhibit elevated expression of the antiapoptosis gene BCL2 and lower expression levels of p21 relative to patients with pN2 disease without skip metastases. Micrometastases occurred in 3 of 17 (17.6%) patients with pN2 disease and skip metastases diagnosed by routine histopathology. PMID- 15112273 TI - Recurrent chromosomal imbalances in nonsmall cell lung carcinoma: the association between 1q amplification and tumor recurrence. AB - BACKGROUND: Lung carcinoma is a leading cause of cancer deaths worldwide. To better understand this disease, the authors studied genetic alterations in nonsmall cell lung carcinoma (NSCLC) and the association between genetic changes and clinical features. METHODS: Genetic alterations in 30 patients with adenocarcinoma (AC) and 39 patients with squamous cell carcinoma (SCC) were analyzed by comparative genomic hybridization. The genetic changes in patients with AC and SCC were compared and the associations of these changes with clinical features were studied. RESULTS: A gain of 3q with a minimal amplified region at 3q25.3-qter was significantly higher in patients with SCC compared with patients with AC (72% vs. 27%; P < 0.001). A gain of 20q and loss of chromosome 9 were detected more frequently in patients with AC compared with patients with SCC (P < 0.05). Gains of 5p and 20q and loss of 5q were significantly correlated with an advanced stage of NSCLC (P < 0.05). Amplification of 1q was significantly associated with NSCLC recurrence (P = 0.04). CONCLUSIONS: The results of the current study suggested that different chromosomal aberrations may contribute to the types and pathologic stages of NSCLC. PMID- 15112274 TI - Outcome after local recurrence of osteosarcoma: the St. Jude Children's Research Hospital experience (1970-2000). AB - BACKGROUND: Despite improvements in therapy for osteosarcoma, approximately 4-10% of patients experience a local recurrence and have a poor prognosis. METHODS: The authors analyzed prognostic factors for survival in 26 patients with a local recurrence of osteosarcoma who were treated between 1970 and 2000. RESULTS: The initial surgical procedure was amputation in 20 patients (76.9%) and limb salvage in 6 patients (23.1%). The median time from the diagnosis of osteosarcoma to local recurrence was 1.2 years (range, 1.2 months-6.1 years). Eleven patients (42.3%) developed an isolated local recurrence and 15 patients (57.7%) developed local and distant recurrence. The 5-year estimate of postrecurrence survival (PRS) (+/- 1 standard error) for the 26 patients was 19.2% +/- 7.7%. Recurrence > or = 2 years from the time of diagnosis was found to predict a better outcome (5 year PRS of 50.0% +/- 20.4%) compared with earlier recurrence (10.0% +/- 5.5%) (P = 0.037). Patients with negative margins after initial surgery were found to have improved survival (5-year PRS of 33.3% +/- 13.6%) compared with patients with positive margins (7.1% +/- 4.9%) (P = 0.015). Patients who underwent complete surgical resection at the time of recurrence were found to have a better PRS (5 year PRS of 41.7% +/- 14.2%) compared with patients who did not undergo surgery (0% +/- 0%) (P < 0.001). CONCLUSIONS: The prognosis for patients after local recurrence of osteosarcoma is poor. Complete surgical resection at the time of recurrence is essential for survival. Positive margins at the time of initial surgical resection and early recurrence appear to be poor prognostic factors. PMID- 15112275 TI - Evaluation of P-glycoprotein, HER-2/ErbB-2, p53, and Bcl-2 in primary tumor and metachronous lung metastases in patients with high-grade osteosarcoma. AB - BACKGROUND: Investigation of the relation between primary tumor and metastatic disease is necessary for the identification of predictive factors for postrecurrence survival (PRS) in patients with recurrent osteosarcoma. METHODS: Cellular levels of P-glycoprotein, ErbB-2, p53, and Bcl-2 expression were evaluated in primary tumor biopsy and metachronous pulmonary metastasis specimens from 19 patients with high-grade osteosarcoma. Results were analyzed for differences between primary tumor and pulmonary metastases and for correlations between expression patterns and survival. RESULTS: Positive staining in lung metastases was noted in 68%, 53%, 32%, and 84% of patients for P-glycoprotein, ErbB-2, p53, and Bcl-2, respectively. These percentages were higher than those observed in primary tumor specimens for all genetic markers evaluated, with a significant difference in the percentage of patients with positive staining for P glycoprotein (68% vs. 32%; P = 0.05) and a near-significant difference in the percentage of patients with positive staining for Bcl-2 (84% vs. 53%; P = 0.08). Patients with ErbB-2 expression in the primary tumor were more likely to have multiple metastases and shorter recurrence-free intervals compared with patients in whom ErbB-2 expression was not observed, whereas differences in P glycoprotein, p53, and Bcl-2 expression were not related to differences in metastatic pattern. PRS was influenced by p53 expression levels in pulmonary metastases, with patients who had negative staining for p53 having a significantly better PRS rate relative to patients with positive staining for p53 (3-year PRS rate: p53-negative, 64%; p53-positive, 17%; P = 0.008). CONCLUSIONS: In the current study of patients with high-grade osteosarcoma, most patients exhibited increased cellular expression of P-glycoprotein, ErbB-2, and Bcl-2 in recurrent pulmonary metastases compared with primary tumor. Further studies aimed at investigating the relation between altered p53 expression in lung metastases and postrecurrence survival are recommended. PMID- 15112276 TI - Hyperthermic intraperitoneal intraoperative chemotherapy after cytoreductive surgery for the treatment of abdominal sarcomatosis: clinical outcome and prognostic factors in 60 consecutive patients. AB - BACKGROUND: Abdominal sarcomatosis is a rare nosologic entity with a poor prognosis. After a Phase I study on cytoreductive surgery combined with hyperthermic intraperitoneal intraoperative chemotherapy (HIIC), the authors reported the results of the treatment of 60 patients using this novel multimodal approach. METHODS: Twenty-nine patients had multifocal primary disease and 31 patients had recurrent abdominal sarcoma. Tumor histology was represented by visceral (n = 26 [43%]) and retroperitoneal (n = 34 [57%]) sarcoma. All patients underwent cytoreductive surgery (with no or minimal residual disease) and 90 minute HIIC with doxorubicin (15.25 mg/L of perfusate) and cisplatin (43 mg/L). The clinical outcome and the prognostic value of 11 clinicopathologic variables were analyzed. RESULTS: No postoperative deaths occurred. The morbidity rate was 33% and the moderate to severe locoregional toxicity rate was 15%. The median time to local disease progression and the median overall survival were 22 months and 34 months, respectively. Using multivariate analysis, histologic grading and completeness of surgical cytoreduction predicted patient prognosis, indicating that both local progression-free and overall survival were affected significantly by tumor aggressiveness and local disease control. CONCLUSIONS: Although these results were encouraging, there was no definitive conclusion reached regarding the therapeutic activity of this locoregional treatment. In addition, the toxicity rate was substantial. In the absence of effective systemic agents, the therapeutic potential of cytoreductive surgery plus HIIC should be explored further in comparative trials. PMID- 15112277 TI - P53 and K-ras mutations are frequent events in microscopically negative surgical margins from patients with nonsmall cell lung carcinoma. AB - BACKGROUND: The objective of the current study was to determine whether tumor cells harboring P53 and K-ras mutations could be detected in histopathologically tumor-free surgical margins in patients with nonsmall cell lung carcinoma who underwent complete pulmonary resection. METHODS: In 118 consecutive patients, DNA obtained from primary tumors and from surgical margins was extracted for molecular analysis. A fragment of P53 gene encompassing exons 5-8 and codon 12 of the K-ras gene were amplified with the polymerase chain reaction technique and were assayed for the presence of mutations. RESULTS: P53 and K-ras mutations were found in 30% and 39% of primary tumors, respectively, and in 11 (9%) and 22 (18%) apparently tumor-free surgical margins, respectively. At least 1 of those mutations was found in surgical margins in 29 patients (25%), and both mutations were found in 2 patients (1.7%). P53 mutations in surgical margins accompanied mutations in primary tumors in 9 of 35 patients (26%), and K-ras mutations accompanied mutations in primary tumors in 20 of 46 patients (44%). Among patients with either mutation in primary tumors, the incidence of at least 1 mutation in surgical margins was 43% (28 of 65 patients). In four patients, mutations (two K-ras mutations and two P53 mutations) were found in surgical margins despite the absence of the corresponding mutations in primary tumors. The presence of mutations in primary tumors and in surgical margins was not related significantly to clinical characteristics or to patient outcomes. CONCLUSIONS: P53 and K-ras mutations are frequent events in surgical margins determined to be tumor free on light microscopy. The clinical relevance of these findings remains to be established. PMID- 15112278 TI - Discrepancies among patients, family members, and physicians in Korea in terms of values regarding the withholding of treatment from patients with terminal malignancies. AB - BACKGROUND: The role of the physician in end-of-life decision-making is complicated. To analyze the controversies that surround therapeutic decision making and the withholding of life-sustaining treatments, the authors compared values regarding therapeutic intervention that were held by physicians and family members of patients with terminal malignancies. METHODS: One hundred fourteen patients with either advanced-stage or terminal disease were enrolled in the current study. Questionnaires were administered to the duty physician and to patients' family members. The questions covered issues such as the use of new anticancer agents with only partial efficacy (15%) and the use of opioid analgesics, intravenous nutrition, feeding tubes, antibiotics, and hemodialysis. In addition, participants were asked about the administration of cardiopulmonary resuscitation (CPR) and the use of ventilators, and when the patient's family consented, the same questionnaire was administered to the patient as well. RESULTS: Seventeen of 114 families refused to answer the questionnaire. Of the 97 available families, only 14 permitted access to the patient. Of those 14 patients, 5 refused to complete the questionnaire. Overall, 100% of families and 87% of patients had some knowledge regarding malignant disease, but only 69% of families and 37% of patients clearly understood the stage of the patient's disease. The use of a new agent with only partial efficacy (approximately 15%) was accepted by 41% of physicians and by 60% of families. The concordance rate between patients' physicians and family members regarding the same patient was 42%. The rankings of the acceptance of treatment by physicians were as follows: opioid analgesics, 100%; antibiotics, 91%; feeding tube, 87%; and intravenous nutrition, 78%. The rankings of the same items by family members were as follows: opioid analgesics, 92%; antibiotics, 89%; intravenous nutrition, 86%; and feeding tube, 75%. The concordance rates between patients' physicians and families were lowest for ventilator application (39%) and CPR (47%). CONCLUSIONS: Values held on issues such as therapeutic decision-making and the withholding of life sustaining treatment for patients with terminal malignancies were discordant between physicians and family members. To resolve controversies regarding the role of the physician in end-of-life decision-making, the values of physicians, patients, and family members should be considered in the final decision-making process. PMID- 15112279 TI - Palliative nutritional intervention in addition to cyclooxygenase and erythropoietin treatment for patients with malignant disease: Effects on survival, metabolism, and function. AB - BACKGROUND: The role of nutrition in the palliative treatment of patients with malignancy-related cachexia is unclear. The goal of the current study was to determine whether specialized, nutrition-focused patient care could improve integrated whole-body metabolism and functional outcome in unselected weight losing patients with malignant disease who were receiving systemic antiinflammatory (cyclooxygenase [COX]-inhibitory) treatment along with erythropoietin (EPO) support. METHODS: Three hundred nine patients with malignant disease who experienced progressive cachexia due to solid tumors (primarily gastrointestinal lesions) were randomized to receive a COX inhibitor (indomethacin, 50 mg twice daily) and EPO (15-40,000 units per week) along with specialized, nutrition-focused patient care (oral nutritional support and home total parenteral nutrition [TPN]) provided on a patient-by-patient basis to attenuate inflammation, prevent anemia, and improve nutritional status. Control patients received the same indomethacin and EPO doses that study patients received without the added nutritional support. All patients were treated and followed until death. Biochemical assays (blood, liver, kidney, and thyroid), nutritional state assessment (food intake and body composition), and exercise testing with simultaneous measurement of whole-body respiratory gas exchange before and during exercise were performed before the start of treatment and then at regular intervals during the treatment period (every 2-30 months after treatment initiation). Statistical analyses were performed on 'intention-to treat' and 'as-treated' bases. RESULTS: Home TPN was provided to approximately 50% of the study patients without severe complications. Over the entire observation period, rhEPO prevented the development of anemia in both study patients and control patients. Intention-to-treat analysis revealed an improvement in energy balance for nutritionally supported patients (P < 0.03); no other significant differences in outcome between study patients and control patients were observed. As-treated analysis demonstrated that patients receiving nutrition experienced prolonged survival (P < 0.01), which was accompanied by improved energy balance (P < 0.001), increasing body fat (P < 0.05), and a greater maximum exercise capacity (P < 0.04). A trend toward increased metabolic efficiency at maximum exercise (P < 0.06) for study patients relative to control patients also was observed. CONCLUSIONS: The results of the current study strongly support that nutrition is a limiting factor influencing survival and that nutritional support protects integrated metabolism and metabolic function in patients with progressive cachexia secondary to malignant disease. PMID- 15112280 TI - Characterization of FAMPAC, a newly identified human pancreatic carcinoma cell line with a hereditary background. AB - BACKGROUND: A novel pancreatic carcinoma cell line, FAMPAC, was identified from investigation of poorly differentiated pancreatic adenocarcinoma cells found in a patient with a familial predisposition to pancreatic carcinoma. A gene responsible for familial pancreatic carcinoma has not been identified to date. METHODS: The FAMPAC cell line was characterized by its morphology, growth rate, tumorigenicity, and chromosomal analysis. Three known tumor suppressor genes, p16/CDKN2, BRCA2, and p53, all of which are important in the development of pancreatic carcinoma and frequently are involved in a variety of cancer syndromes, were analyzed. RESULTS: FAMPAC cells grew as an adhering monolayer in culture medium supplemented with 10% fetal bovine serum and formed tumors rapidly in nude mice. The doubling time ranged from 24 to 48 hours. Karyotype analysis demonstrated the complexity of chromosomal deletions and rearrangements. The cells were negative for ductal differentiation markers such as cytokeratin 7 and MUC1, indicating poor differentiation. Analysis of FAMPAC cells revealed overexpression of the mutated p53 gene (exon 5, codon 175: CGC --> CAC), the presence of a homozygous deletion in the p16 gene, and the presence of wild-type BRCA2 in the tested hot spots. CONCLUSIONS: To the authors' knowledge, FAMPAC is the first established human pancreatic carcinoma cell line associated with a familial background. FAMPAC is a tumorigenic cell line with a complex molecular pattern of mutations. These findings may be useful in understanding the mechanisms responsible for the development of sporadic or hereditary forms of pancreatic carcinoma. PMID- 15112281 TI - Zoledronic acid delays the onset of skeletal-related events and progression of skeletal disease in patients with advanced renal cell carcinoma. PMID- 15112283 TI - Predictive modeling for the presence of prostate carcinoma using clinical, laboratory, and ultrasound parameters in patients with prostate-specific antigen levels < or = 10 ng/ml. PMID- 15112290 TI - Bioprocess design and economic analysis for the commercial production of environmentally friendly bioinsecticides from Bacillus thuringiensis HD-1 kurstaki. AB - A production process for B. thuringiensis (Bt) bioinsecticides was designed in detail, including alternative batch, low-density fed-batch (LDFB), and high density fed-batch (HDFB) fermentation configurations. Capital and operating costs, as well as profitability based on simple rate of return, were performed using a purpose-written FORTRAN program, explicitly analyzing production of a water-based flowable product used in forestry applications. The total capital cost was 18 million dollars (Canadian dollars) for a stand-alone plant with base scale capacity of 3 x 10(7) billion international units (BIU)/year. Raw material costs amounted to 1.5 million dollars yearly, of which approximately half was for formulation ingredients. Per-unit production cost rose sharply for scales of less than 1 x 10(7) BIU/year, but was little affected by scale above 3 x 10(7) BIU/year. Product cost was much lower at all scales for a LDFB as opposed to batch fermentation process, but HDFB gave relatively little additional cost benefit. Profitability analysis performed by co-varying scale and selling price showed that break-even occurred at a price of 0.45 dollars/BIU for a batch process at base scale, while with LDFB fermentation the same production volume sold at 0.35 dollars/BIU gave a 12% rate of return. Since the assumed base scale would represent 8-15% of current world Bt bioinsecticide production, based on value or volume, it was concluded that profitability would require some or all of the following elements: targeting higher-value markets such as disease vector control, in addition to forestry; a potentially lower plant capacity (although at least 1 x 10(7) BIU/year;) and coproduction of other large-volume microbial products to absorb capacity and match bioinsecticide output to market demand. PMID- 15112291 TI - Quantifying anisotropic solute transport in protein crystals using 3-D laser scanning confocal microscopy visualization. AB - The diffusion of a solute, fluorescein into lysozyme protein crystals has been studied by confocal laser scanning microscopy (CLSM). Confocal laser scanning microscopy makes it possible to non-invasively obtain high-resolution three dimensional (3-D) images of spatial distribution of fluorescein in lysozyme crystals at various time steps. Confocal laser scanning microscopy gives the fluorescence intensity profiles across horizontal planes at several depths of the crystal representing the concentration profiles during diffusion into the crystal. These intensity profiles were fitted with an anisotropic model to determine the diffusivity tensor. Effective diffusion coefficients obtained range from 6.2 x 10(-15) to 120 x 10(-15) m2/s depending on the lysozyme crystal morphology. The diffusion process is found to be anisotropic, and the level of anisotropy depends on the crystal morphology. The packing of the protein molecules in the crystal seems to be the major factor that determines the anisotropy. PMID- 15112292 TI - Enzymatic labeling of a single chain variable fragment of an antibody with alkaline phosphatase by microbial transglutaminase. AB - Functional cross-linking of a single chain Fv fragment of anti-hen egg-white lysozyme antibody (scFv) and alkaline phosphatase (AP) was explored using microbial transglutaminase (MTG) from Streptomyces mobaraensis. A specific peptidyl linker for MTG was genetically fused to the N-terminus of each protein and the resultant proteins were obtained separately by bacterial expression. The recombinant peptide-tagged scFv and AP were site-specifically cross-linked by MTG through the extra peptidyl linkers in vitro, which mainly yielded the heterodimer (i.e., scFv-AP conjugate). The enzymatic cross-linking reaction had little influence on either the antigen-binding ability of the scFv moiety or the enzymatic activity of the AP moiety of the conjugate, allowing use within an enzyme-linked immunosorbent assay. The results obtained suggest that the enzymatic approach with MTG facilitates the posttranslational construction of functional fusion proteins. PMID- 15112293 TI - Combined discrete particle and continuum model predicting solid-state fermentation in a drum fermentor. AB - The development of mathematical models facilitates industrial (large-scale) application of solid-state fermentation (SSF). In this study, a two-phase model of a drum fermentor is developed that consists of a discrete particle model (solid phase) and a continuum model (gas phase). The continuum model describes the distribution of air in the bed injected via an aeration pipe. The discrete particle model describes the solid phase. In previous work, mixing during SSF was predicted with the discrete particle model, although mixing simulations were not carried out in the current work. Heat and mass transfer between the two phases and biomass growth were implemented in the two-phase model. Validation experiments were conducted in a 28-dm3 drum fermentor. In this fermentor, sufficient aeration was provided to control the temperatures near the optimum value for growth during the first 45-50 hours. Several simulations were also conducted for different fermentor scales. Forced aeration via a single pipe in the drum fermentors did not provide homogeneous cooling in the substrate bed. Due to large temperature gradients, biomass yield decreased severely with increasing size of the fermentor. Improvement of air distribution would be required to avoid the need for frequent mixing events, during which growth is hampered. From these results, it was concluded that the two-phase model developed is a powerful tool to investigate design and scale-up of aerated (mixed) SSF fermentors. PMID- 15112294 TI - Morphologically structured model for antitumoral retamycin production during batch and fed-batch cultivations of Streptomyces olindensis. AB - A morphologically structured model is proposed to describe trends in biomass growth, substrate consumption, and antitumoral retamycin production during batch and fed-batch cultivations of Streptomyces olindensis. Filamentous biomass is structured into three morphological compartments (apical, subapical, and hyphal), and the production of retamycin, a secondary metabolite, is assumed to take place in the subapical cell compartment. Model accounts for the effect of glucose as well as complex nitrogen source on both the biomass growth and retamycin production. Laboratory data from bench-scale batch and fed-batch fermentations were used to estimate some model parameters by nonlinear regression. The predictive capability of the model was then tested for additional fed-batch and continuous experiments not used in the previous fitting procedure. The model predictions show fair agreement to the experimental data. The proposed model can be useful for further studies on process optimization and control. PMID- 15112295 TI - Long-term continuous monitoring of dissolved oxygen in cell culture medium for perfused bioreactors using optical oxygen sensors. AB - For long-term growth of mammalian cells in perfused bioreactors, it is essential to monitor the concentration of dissolved oxygen (DO) present in the culture medium to ascertain the health of the cells. An optical oxygen sensor based on dynamic fluorescent quenching was developed for long-term continuous measurement of DO for NASA-designed rotating perfused bioreactors. Tris(4,7-diphenyl-1,10 phenanthroline) ruthenium(II) chloride is employed as the fluorescent dye indicator. A pulsed, blue LED was chosen as the excitation light source. The sensor can be sterilized using an autoclave. The sensors were tested in a perfused rotating bioreactor supporting a BHK-21 (baby hamster kidney) cell culture over one 28-day, one 43-day, and one 180-day cell runs. The sensors were initially calibrated in sterile phosphate-buffered saline (PBS) against a blood gas analyzer (BGA), and then used continuously during the entire cell culture without recalibration. In the 180-day cell run, two oxygen sensors were employed; one interfaced at the outlet of the bioreactor and the other at the inlet of the bioreactor. The DO concentrations determined by both sensors were compared with those sampled and measured regularly with the BGA reference. The sensor outputs were found to correlate well with the BGA data throughout the experiment using a single calibration, where the DO of the culture medium varied between 25 and 60 mm Hg at the bioreactor outlet and 80-116 mm Hg at the bioreactor inlet. During all 180 days of culture, the precision and the bias were +/-5.1 mm Hg and -3.8 mm Hg at the bioreactor outlet, and +/- 19 mm Hg and -18 mm Hg at inlet. The sensor dynamic range is between 0 and 200 mm Hg and the response time is less than 1 minute. The resolution of the sensor is 0.1 mm Hg at 50 mm Hg, and 0.25 mm Hg at 130 mm Hg. PMID- 15112296 TI - Epitope mapping and features of the epitope for monoclonal antibodies inhibiting enzymatic activity of Helicobacter pylori urease. AB - Two characteristic monoclonal antibodies (HpU-2 and -18) out of 26 monoclonal antibodies (HpU-1 approximately 26) produced against Helicobacter pylori (H. pylori) urease showed a strong inhibitory effect against the enzymatic activity of the urease. Epitope mapping about some monoclonal antibodies of the HpU-series inhibiting enzymatic activity was performed by using a surface plasmon resonance apparatus and by digesting H. pylori urease with trypsin, followed by mass spectroscopy. The sequences of the epitopes recognized by HpU-2 and -18 were SVELIDIGGNRRIFGFNALVDR (22 mer) and IFGFNALVDR (10 mer), respectively. The former sequence is present as a part of a loop structure at a position close to the C terminal of the alpha-subunit of H. pylori urease, although it has been suggested that the active site of the urease resides in the beta-subunit. The above peptide (22 mer) was chemically synthesized in a linear and cyclic form, and its conjugate with BSA was immunized in rabbits. The resultant serum induced by the linear form could specifically bind to H. pylori infecting human gastric mucosa. These results suggest that the above sequence (22 mer) must be an important epitope, although it locates in the alpha-subunit but not in the beta-subunit. PMID- 15112297 TI - Determination of external and internal mass transfer limitation in nitrifying microbial aggregates. AB - In this article we present a study of the effects of external and internal mass transfer limitation of oxygen in a nitrifying system. The oxygen uptake rates (OUR) were measured on both a macro-scale with a respirometric reactor using off gas analysis (Titrimetric and Off-Gas Analysis (TOGA) sensor) and on a micro scale with microsensors. These two methods provide independent, accurate measurements of the reaction rates and concentration profiles around and in the granules. The TOGA sensor and microsensor measurements showed a significant external mass transfer effect at low dissolved oxygen (DO) concentrations in the bulk liquid while it was insignificant at higher DO concentrations. The oxygen distribution with anaerobic or anoxic conditions in the center clearly shows major mass transfer limitation in the aggregate interior. The large drop in DO concentration of 22-80% between the bulk liquid and aggregate surface demonstrates that the external mass transfer resistance is also highly important. The maximum OUR even for floccular biomass was only attained at much higher DO concentrations (approximately 8 mg/L) than typically used in such systems. For granules, the DO required for maximal activity was estimated to be >20 mg/L, clearly indicating the effects of the major external and internal mass transfer limitations on the overall biomass activity. Smaller aggregates had a larger volumetric OUR indicating that the granules may have a lower activity in the interior part of the aggregate. PMID- 15112298 TI - Methanol induction optimization for scFv antibody fragment production in Pichia pastoris. AB - Fibronectin splice variant ED B (extracellular domain B) is a promising marker for angiogenesis in growing solid tumors. Currently, recombinant antibodies against ED B are being investigated concerning their potential use, for either therapeutic or diagnostic purposes. Single-chain antibody fragments directed against the ED B can be efficiently expressed in Pichia pastoris; thus, a recombinant strain of the methylotropic yeast P. pastoris was used for this work. Three different forms of scFv antibody fragment are found in the supernatant from this fermentation: covalent homodimer, associative homodimer, and monomer. Both homodimeric forms can be converted to the monomeric form (under reducing conditions) and be efficiently radiolabeled, whereas the monomeric form of scFv already present in the supernatant cannot. It was also found that the fraction of protein in the monomeric form is highly dependent on the mode of induction rather than scFv concentration. This suggests that the monomeric form of the scFv present in the supernatant might be a result of events occurring at the expression, secretion, or folding level. A high cell density fermentation protocol was developed by optimizing methanol induction, yielding the highest scFv antibody fragment production rate and product quality; cell concentration at the induction point and specific methanol uptake rate were found to be the most important control variables. A decrease in specific methanol uptake rate led to a higher specific production rate for the scFv antibody fragment (5.4 microg g(cell) h(-1)). Product quality, i.e., percentage of product in a homodimeric form, also increased with the decrease in methanol uptake rate. Furthermore, the volumetric productivity depended on cell concentration at the induction point, increasing with the increase of cell concentration up to 320 g L(-1) wet cell weight (WCW). The reduction of the methanol feeding rate for induction, and consequently of the oxygen uptake rate, have important consequences for optimizing product titers and quality and thus on the scale-up of this production process; hence one of the major limitations upon high cell density cultivation in bioreactors is keeping the high oxygen transfer rate required. From the results obtained, a scale-up strategy was developed based on the available oxygen transfer rates at larger scales, allowing the definition of the optimum biomass concentration for induction and methanol feeding strategy for maximization of product titer and quality. PMID- 15112299 TI - Treatment of MEK and toluene mixtures in biofilters: effect of operating strategy an performance during transient loading. AB - In recent years, biofiltration has been increasingly applied as an air pollution control technology to minimize or eliminate emissions of volatile organic compounds from industrial sources and environmental remediation activities. Although the ability of this technology to maintain high removal efficiency during relatively steady loading conditions has been well established for many waste streams, relatively little research has focused on development of operating strategies that could improve treatment performance during transient loading conditions typical of industrial operations. In the research described herein, two operating strategies were evaluated over a period of 295 days in biofilters treating a model waste gas stream containing a two-component mixture of methyl ethyl ketone (MEK) and toluene. One biofilter was operated as a sequencing batch biofilter (SBB), and the other was operated as a conventional continuous-flow biofilter (CFB). During "normal" steady loading conditions, the model waste stream contained MEK concentrations ranging from 80 to 89 ppmv and toluene concentrations ranging from 28 to 30 ppmv. Both biofilter operating strategies resulted in stable long-term performance with greater than 99% contaminant removal during these normal loading conditions. On a regular basis, the influent MEK and toluene concentrations were temporarily increased to five times the normal influent concentration for the duration of 1 h to test performance during transient "shock loading" conditions. Biofilter performance during the model shock loading conditions demonstrate that SBB operating strategies can result in superior treatment in two important areas: (1) overall mass of contaminants removed and (2) minimum instantaneous removal efficiency. PMID- 15112300 TI - The 2003 Nobel Prize for MRI: significance and impact. PMID- 15112301 TI - A comparison of Ktrans measurements obtained with conventional and first pass pharmacokinetic models in human gliomas. AB - PURPOSE: To compare in a group of patients with cerebral gliomas the estimates of Ktrans between a conventionally established pharmacokinetic model and a recently developed first pass method. MATERIALS AND METHODS: Glioma patients (23) were studied using T1-weighted dynamic contrast-enhanced magnetic resonance imaging (MRI), and two alternative pharmacokinetic models were used for analysis to derive the volume transfer constant Ktrans. These were a modified version of the established model (yielding KTK) and a recently published method based on first pass leakage profile (FP) of contrast bolus (yielding Kfp). RESULTS: We found a strong correlation between intra-tumoral median KTK and Kfp (rho = 0.650, P < 0.01), but the values from the conventional model were consistently and significantly higher (mean of inter-tumoral Kfp and KTK medians were 0.018 minute(-1) and 0.284 minute(-1), respectively, P < 0.001). The spatial distribution of KTK and Kfp showed poor correlation in the presence of large vascular structures and good correlation elsewhere. CONCLUSION: KTK and Kfp produce similar biologic information within voxels not dominated by vascular tissue. The FP method avoids erroneous overestimation of Ktrans in areas of significant intravascular contrast. Findings are in keeping with the predictions of previous mathematical simulations. PMID- 15112302 TI - 1H metabolite relaxation times at 3.0 tesla: Measurements of T1 and T2 values in normal brain and determination of regional differences in transverse relaxation. AB - PURPOSE: To measure 1H relaxation times of cerebral metabolites at 3 T and to investigate regional variations within the brain. MATERIALS AND METHODS: Investigations were performed on a 3.0-T clinical whole-body magnetic resonance (MR) system. T2 relaxation times of N-acetyl aspartate (NAA), total creatine (tCr), and choline compounds (Cho) were measured in six brain regions of 42 healthy subjects. T1 relaxation times of these metabolites and of myo-inositol (Ins) were determined in occipital white matter (WM), the frontal lobe, and the motor cortex of 10 subjects. RESULTS: T2 values of all metabolites were markedly reduced with respect to 1.5 T in all investigated regions. T2 of NAA was significantly (P < 0.001) shorter in the motor cortex (247 +/- 13 msec) than in occipital WM (301 +/- 18 msec). T2 of the tCr methyl resonance showed a corresponding yet less pronounced decrease (162 +/- 16 msec vs. 178 +/- 9 msec, P = 0.021). Even lower T2 values for all metabolites were measured in the basal ganglia. Metabolite T1 relaxation times at 3.0 T were not significantly different from the values at 1.5 T. CONCLUSION: Transverse relaxation times of the investigated cerebral metabolites exhibit an inverse proportionality to magnetic field strength, and especially T2 of NAA shows distinct regional variations at 3 T. These can be attributed to differences in relative WM/gray matter (GM) contents and to local paramagnetism. PMID- 15112303 TI - Survival analysis in patients with glioblastoma multiforme: predictive value of choline-to-N-acetylaspartate index, apparent diffusion coefficient, and relative cerebral blood volume. AB - PURPOSE: To investigate the potential value of pre-external-beam radiation therapy (XRT) choline-to-NAA (N-acetylaspartate) index (CNI), apparent diffusion coefficient (ADC), and relative cerebral blood volume (rCBV) for predicting survival in newly diagnosed patients with glioblastoma multiforme (GBM). MATERIALS AND METHODS: Twenty-eight patients with GBM were studied using in vivo proton magnetic resonance spectroscopic imaging (1H MRSI) and diffusion- and perfusion-weighted imaging after surgery but prior to XRT. Patients were categorized on the basis of their volumes of morphologic and metabolic abnormalities (volume of CNI > or = 2 and CNI values), normalized ADC (nADC), or rCBV values within the T1 contrast-enhancing and T2 regions. The median survival time was compared. RESULTS: A significantly shorter median survival time was observed for patients with a large volume of metabolic abnormality than for those with a small abnormality (12.0 and 17.1 months, respectively, P = 0.002). A similar pattern was observed for patients with a low mean nADC value compared to those with high mean nADC value within the T2 region (11.2 and 21.7 months, respectively, P = 0.004). A shorter median survival time was also observed for patients with contrast-enhancing residual disease than for those without the presence of contrast enhancement with marginal significance. CONCLUSION: The pre XRT volume of the metabolic abnormality and the nADC value within the T2 region may be valuable in predicting outcome for patients with GBM. PMID- 15112304 TI - Multislice first-pass myocardial perfusion imaging: Comparison of saturation recovery (SR)-TrueFISP-two-dimensional (2D) and SR-TurboFLASH-2D pulse sequences. AB - PURPOSE: To compare signal-to-noise ratio (SNR), contrast-to-noise (CNR) ratio, and diagnostic accuracy of a newly developed saturation recovery (SR)-TrueFISP two-dimensional (2D) sequence with an SR-TurboFLASH-2D sequence. MATERIALS AND METHODS: In seven healthy subjects and nine patients with coronary artery disease (CAD), contrast-enhanced perfusion imaging (with Gd-DTPA) was performed with SR TrueFISP and SR-TurboFLASH sequences. Hypoperfused areas were assessed qualitatively (scale = 0-4). Furthermore, SNR and CNR were calculated and semiquantitative perfusion parameters were determined from signal intensity (SI) time curves. Standard of reference for patient studies was single-photon emission computer tomography (SPECT) and angiography. RESULTS: The perception of perfusion deficits was superior in TrueFISP images (2.6 +/- 1.0) than in TurboFLASH (1.4 +/ 0.6) (P < 0.001). Phantom measurements yielded increased SNR (143 +/- 34%) and CNR (158 +/- 64%) values for TrueFISP. In patient/volunteer studies SNR was 61% to 100% higher and signal enhancement was 110% to 115% higher with TrueFISP than with TurboFLASH. Qualitative and semiquantitative assessment of perfusion defects yielded higher sensitivities for detection of perfusion defects with TrueFISP (68% to 78%) than with TurboFLASH (44% to 59%). CONCLUSION: SR-TrueFISP-2D perfusion imaging provides superior SNR and CNR than TurboFLASH imaging. Moreover, the dynamic range of SIs was found to be higher with TrueFISP, resulting in an increased sensitivity for detection of perfusion defects. PMID- 15112305 TI - Uptake of MnCl2 and mangafodipir trisodium in the myocardium: a magnetic resonance imaging study in pigs. AB - PURPOSE: To examine the changes in the longitudinal relaxation times (DeltaR1) induced in pig myocardium and blood following injections of 5, 10, and 15 micromol mangafodipir trisodium (Mn-DPDP) or MnCl2/kg of body weight (b.w.). MATERIALS AND METHODS: Twelve pigs were divided into two groups, one group receiving MnCl2 and the other receiving Mn-DPDP. Three consecutive doses of contrast agent (5, 10, and 15 micromol/kg of b.w.) were injected in each animal with a 40-minute time interval between each dose. Measurements of T1 in blood and myocardium were made 5, 15, 25, and 35 minutes after each injection. Additionally, relaxivity measurements in blood samples were performed. RESULTS: An increase in myocardial R1 was observed for both contrast agents at all concentration levels tested. This increase peaked 5 minutes after injection and then declined. An increase could still be detected 35 minutes after injection. The effect was larger when using MnCl2 than when using Mn-DPDP. CONCLUSION: The dissociation kinetics of Mn2+ from the DPDP ligand limits the relaxation increase of Mn-DPDP relative to that of MnCl2. On the other hand, the toxicity of MnCl2 may exclude it from clinical use. PMID- 15112306 TI - In vivo mapping of spontaneous mammary tumors in transgenic mice using MRI and ultrasonography. AB - PURPOSE: To compare T1- or T2-weighted magnetic resonance imaging (MRI) and ultrasonography (US) as tools for in vivo mapping of different tissue components in spontaneous tumors of transgenic mice. MATERIALS AND METHODS: Human-like mammary adenocarcinomas from FVB/neuT transgenic mice were analyzed by T2 weighted and T1-weighted MRI at 4.7 Tesla and US and then, after excision, were paraffin-embedded for histologic analysis. The histologic samples were prepared taking care to obtain sections that spatially matched the MRI and US images as precisely as possible. RESULTS: US can obtain basic information such as the size of developing tumors in experimental animals and can identify necrotic areas. T2 weighted MRI, especially if compared to T1-weighted MRI and/or US, allows advanced analysis of morphologic aspects, with high resolution in the differentiation of details of necrotic areas such as coagulation, liquefaction, biphasic splitting of cysts, and fibrotic and lipidic infiltration. CONCLUSION: Of the three methods, T2-weighted MRI provides the most information about the anatomy of tumors. However, when distinctions between the different types of necrosis are not needed, US analysis is to be preferred for its practicality. PMID- 15112307 TI - MR-guided intravascular procedures: real-time parameter control and automated slice positioning with active tracking coils. AB - PURPOSE: To implement and optimize a real-time pulse sequence and user interface to perform intravascular interventions using active catheter tracking. MATERIALS AND METHODS: In magnetic resonance (MR)-guided interventions, small radio frequency coils can be used to rapidly determine the device position (active tracking). In this work, active catheter tracking was combined with a dedicated real-time pulse sequence and user interface. The pulse sequence offered the imaging contrasts fast low angle shot (FLASH), true Fast imaging with steady state precession (TrueFISP), and projection MR digital subtraction angiography (MR-DSA), which could be selected by the radiologist from within the scanner room at any time during the intervention. Automatic slice positioning was added to the real-time pulse sequence so that the location of the tracking coils defined the image slice position and orientation. The technique was assessed in phantoms and animal experiments. RESULTS: At a reaction time of 24 msec and a frame rate of three images per second, the movement of an active intravascular catheter could be monitored in the aorta and the renal arteries of a pig. With interactive contrast and orientation changes, the renal vasculature could be assessed by a fully MR-guided catheterization in less than 10 minutes. CONCLUSION: With carefully designed active catheters, a dedicated user interface, and an optimized pulse sequence intravascular interventions can successfully be performed by a single operator from within the MR scanner room. PMID- 15112308 TI - Soft tissue enhancement on time-resolved peripheral magnetic resonance angiography. AB - PURPOSE: To evaluate the incidence and locations of soft tissue enhancement on time-resolved two-dimensional projection magnetic resonance angiography (MRA) of the calf and foot. MATERIALS AND METHODS: Time-resolved two-dimensional projection MRA of the knee, calf, and foot, performed at 1.5 Tesla using the head coil, was retrospectively reviewed in 326 patients. Soft tissue enhancement of the foot was identified and graded by blinded review. Subsequently, patient medical records were reviewed to determine the presence or absence of diabetes, cellulitis, gangrene, and ulceration of the foot and presence of neuropathic joints. RESULTS: Enhancing spots were identified in 228 patients. For those feet without clinical abnormalities, diabetic patients had an average of 1.2 enhancing spots per foot, while nondiabetics had only 0.6 (P < 0.001). Higher-grade lesions were more prevalent in diabetics and in heavier patients. Of 64 patients with follow-up, 8 (13%) developed cellulitis (N = 4) or ulceration (N = 4) at the location of an enhancing spot, including 5 diabetic and 3 nondiabetic patients. CONCLUSION: Pedal soft tissue enhancement frequently occurs on time-resolved gadolinium (Gd):MRA of the feet. The etiology is uncertain, but the high frequency in diabetic patients and observation of progression to cellulitis/ulceration suggest this soft tissue enhancement may identify sites of subclinical pedal soft tissue injury. PMID- 15112309 TI - In vivo proton magnetic resonance spectroscopy of large focal hepatic lesions and metabolite change of hepatocellular carcinoma before and after transcatheter arterial chemoembolization using 3.0-T MR scanner. AB - PURPOSE: To investigate the value of in vivo proton magnetic resonance spectroscopy (MRS) in the assessment of large focal hepatic lesions and to measure the metabolite change of hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE) using 3.0-T scanner. MATERIALS AND METHODS: In this prospective study, 43 consecutive patients with large (not less than 3 cm in diameter) hepatic tumors and eight normal volunteer were included. MRS of the lesions in addition to uninvolved liver parenchyma was carried out using a whole-body 3.0-T scanner. Among the patients with proven HCC, eight lesions were evaluated before and two to five days after TACE. The choline to-lipid (cho/lipid) ratio was measured by dividing the peak area of choline at 3.2 ppm by the peak area of lipid at 1.3 ppm. The sensitivity and specificity profiles of MRS in the diagnosis of malignant hepatic tumors were determined by plotting empirical receiver operating characteristic (ROC) curve. The mean cho/lipid ratios in different groups before and after TACE were also measured. RESULTS: The technical success rate for MRS was 90% (53/59). The ROC curve showed proton MRS has moderate discriminating ability in diagnosing malignant hepatic tumors, although the sensitivity was less than 50% while 1-specificity was less than 20%. The area under the curve was 0.71 (P < 0.05). The mean +/- 1 standard error (SE) of cho/lipid ratios for uninvolved liver (N = 8), benign tumor (N = 8), and malignant tumor (N = 21; 19 HCC, one angiosarcoma, and one lymphoma) were 0.06 +/- 0.02, 0.02 +/- 0.02, and 0.17 +/- 0.05, respectively. A significantly statistical difference (ANOVA planned contrast test, P = 0.01 and Games-Howell procedure, P = 0.03) was achieved in the mean cho/lipid ratio between malignant and benign tumors. The mean cho/lipid ratios were significantly decreased from 0.23 +/- 0.11 before TACE to 0.01 +/- 0.00 after the treatment (t = 2.01, P < 0.05, one-tail paired t-test; z = -2.37, P < 0.05, Wilcoxon Signed Ranks Test). CONCLUSION: In vivo proton MRS is technically feasible for the evaluation of focal hepatic lesions. The technique has potential in the detection of early metabolite change in malignant liver tumors after TACE but limitation still exists in clear differentiation between normal liver and benign and malignant tumor. PMID- 15112310 TI - Diaphragmatic motion in the sitting and supine positions: Healthy subject study using a vertically open magnetic resonance system. AB - PURPOSE: To determine the postural difference of diaphragmatic motion between the sitting and supine positions. MATERIALS AND METHODS: A total of 10 healthy men were examined using a vertically open 0.5-T magnetic resonance (MR) system. A total of 40 sequential MR images were obtained in both the sitting and supine positions during two to five respiratory cycles. The diaphragmatic excursions (DEs) were measured on three diaphragmatic points of six sagittal planes for both positions. The differences in DEs between the anterior and posterior parts of the diaphragm were also determined. RESULTS: DEs in the supine position were significantly greater than those in the sitting position at 15 of the 18 points. In five of the six sagittal planes, the difference of DE between posterior and anterior points was significantly larger in the supine position than in the sitting position. CONCLUSION: Diaphragmatic movement in the supine position is greater than that in the sitting position, especially in the posterior part of the diaphragm. PMID- 15112311 TI - Evaluation of the reproducibility of intrarenal R2* and DeltaR2* measurements following administration of furosemide and during waterload. AB - PURPOSE: To estimate the reproducibility of BOLD MRI measurements in the evaluation of intrarenal oxygenation levels. MATERIALS AND METHODS: In this study, the reproducibility of semiquantitative BOLD MRI measurements performed on a 1.5 T scanner with a multiple gradient-echo sequence in the renal medulla and cortex, and their response to furosemide and waterload, were assessed in eight healthy young subjects (25.6 +/- 4.1 years). Each subject underwent an identical experimental procedure on two separate days. RESULTS: Renal R2* measurements were shown to be reproducible within approximately 12% from day to day based on a coefficient of variance (CV) analysis. The changes in R2* (DeltaR2*) following administration of furosemide were statistically significant, as shown by ANOVA and a paired Student's t-test, and were deemed reliable based on the reliable change index (RCI). However, DeltaR2* values following waterload were not statistically significant, and were not deemed reliable. CONCLUSION: R2* measurements were reproducible over 270 days within 12%. Furosemide produced a significant and reliable change (approximately 30%), and the magnitude of change (5.7 s(-1)) was reproducible and consistent with our previous data. The response to waterload, however, did not reach statistical significance, and the magnitude did not reach the level that we had previously reported. PMID- 15112312 TI - Significant differences in proton trimethyl ammonium signals between human gastrocnemius and soleus muscle. AB - PURPOSE: To study the apparent heterogeneous characteristics of trimethyl ammonium (TMA) in healthy human muscles at rest, and to illustrate the importance of establishing the baseline characteristics of proton metabolites in muscles with a West Nile patient. MATERIALS AND METHODS: Point-resolved spectroscopy (PRESS) magnetic resonance spectroscopy imaging (MRSI) with lipid suppression and optional outer-volume presaturation were used to acquire 1H spectra of human muscles at rest at 1.5 Tesla. A total of 28 subjects (27 normal volunteers and 1 patient with West Nile disease) between the ages of 22 and 76 participated in the study. RESULTS: The apparent T2 values of TMA for soleus and gastrocnemius muscles in normal volunteers are 180 +/- 50 and 80 +/- 20 msec, respectively. This difference has profound effects on the apparent spectral pattern of 1H metabolites. The TMA/total creatine (tCr) spectral pattern of the soleus muscle of a West Nile patient resembles that of gastrocnemius muscle of healthy volunteers. CONCLUSION: There are significant differences in the apparent T2 values of TMA between healthy soleus and gastrocnemius muscles at rest. It is important to establish the baseline characteristics of proton metabolites before clinical or physiological studies can be performed. PMID- 15112313 TI - Assessment of the rotator cuff and glenoid labrum using an extremity MR system: MR results compared to surgical findings from a multi-center study. AB - PURPOSE: To conduct a multi-center assessment of the use of a 0.2-T, extremity MR system (E-scan; General Electric Lunar Corp. and Esaote, Genoa, Italy) for identifying tears of the rotator cuff and glenoid labrum. MATERIALS AND METHODS: A retrospective study was performed involving 160 patients (age range, 15-84 years old) from five facilities in the United States, comparing shoulder MR imaging to surgical findings. MR imaging of the shoulder was conducted as follows: shoulder coil; T1-weighted spin echo, coronal-oblique and axial images; short Tau inversion recovery (STIR), coronal-oblique images and axial images; and T2-weighted spin echo, coronal-oblique and sagittal-oblique images. The MR examinations were interpreted in an independent, prospective manner by two radiologists with extensive musculoskeletal MRI experience. Arthroscopic (N = 103) or open surgical (N = 57) procedures were performed within a mean of 53 days after MR imaging. RESULTS: Surgical findings demonstrated rotator cuff tears in 131 patients and labral tears in 60 patients. For the rotator cuff, the sensitivity, specificity, positive predictive value, and negative predictive value were 90%, 93%, 98%, and 68%, respectively. For the labrum, the sensitivity, specificity, positive predictive value, and negative predictive value were 55%, 100%, 100%, and 82%, respectively. CONCLUSIONS: There was good agreement when MR results obtained using the extremity MR system were compared to surgical findings for identifying rotator cuff tears, while the sensitivity of MR imaging for determining labral tears was relatively poor. Nevertheless, these findings were comparable to those reported in the peer-reviewed literature for MR systems operating at mid-, and high-field-strengths. PMID- 15112314 TI - Noise reduction in MR angiography with nonlinear anisotropic filtering. AB - PURPOSE: To evaluate three-dimensional nonlinear anisotropic filtering in suppressing image noise in high spatial resolution magnetic resonance angiograms (MRA) acquired with hybrid undersampled projection reconstruction and phase contrast vastly undersampled isotropic projection reconstruction (PC-VIPR). MATERIALS AND METHODS: Three-dimensional nonlinear anisotropic filtering was quantitatively analyzed and evaluated through the measurement of contrast to noise ratio (CNR) in PC-VIPR images and contrast enhanced peripheral MRA images. To filter MRA images with ultra-high spatial resolution and poor CNR, a spatial frequency dependent nonlinear anisotropic filtering algorithm was proposed that uses two-step processing to filter the whole spatial frequency data. RESULTS: Three-dimensional nonlinear anisotropic filtering was shown to be effective in suppressing noise and improving CNR in MRA with isotropic spatial resolution. Higher CNR was achieved using spatial frequency dependent nonlinear anisotropic filtering. A typical CNR gain of between 50-100% was shown in our studies. CONCLUSION: Three-dimensional nonlinear anisotropic filtering significantly improved CNR in MRA images with isotropic spatial resolution. Spatial frequency dependent nonlinear anisotropic filtering further improved CNR for MRA images with ultra-high spatial resolution and low CNR. PMID- 15112315 TI - Pheochromocytoma with posthemorrhagic cystic degeneration: magnetic resonance imaging findings. AB - We describe in vivo and in vitro magnetic resonance imaging (MRI) findings of a pheochromocytoma with posthemorrhagic cystic degeneration in a 74-year-old man. The in vivo MR images showed the mass as an area of homogeneous moderate hyperintensity with a central area of intense hyperintensity outlined by a thin hypointense rim on a T2-weighted image. The in vitro MR images showed a hyperintense rim around the central cystic area consistent with hemorrhage on T1 weighted gradient-echo images with short echo times (1.6 and 4.2 msec) and more distinctly revealed the blurring effect due to susceptibility of hemosiderin on those with long echo times (6 and 8 msec). Hemosiderin deposition caused by intraparenchymal hemorrhage in the pheochromocytoma can be appreciated on spoiled gradient-echo images with different echo times, which is ordinarily included in the MRI protocol as phase-shift imaging. PMID- 15112316 TI - Coronary artery imaging using three-dimensional breath-hold steady-state free precession with two-dimensional iterative partial fourier reconstruction. AB - PURPOSE: To assess the feasibility of using a two-dimensional partial Fourier (PF) reconstruction scheme to reduce the acquisition time of magnetic resonance imaging (MRI) of coronary arteries. MATERIALS AND METHODS: Symmetric k-space data sets of coronary arteries were collected in seven volunteers using a three dimensional breath-hold steady-state free precession (SSFP) sequence. Partial, asymmetric k-space data sets were generated by removing 25% of the data in the readout direction and 25% of the data in the phase encoding direction. The missing data were then estimated using a two-dimensional projection-onto-convex sets (POCS) algorithm or filled with zeroes. Images were reconstructed from the full data set, the PF data set, and the zero-filled (ZF) data set, respectively. Coronary artery sharpness was evaluated quantitatively and qualitatively. RESULTS: Coronary artery sharpness in PF images was comparable to that in full k space images and significantly better than that in ZF images. CONCLUSION: Two dimensional POCS PF reconstruction is a potentially useful technique for reducing acquisition time or improving spatial resolution for breath-hold coronary MR angiography. PMID- 15112317 TI - Temperature and SAR calculations for a human head within volume and surface coils at 64 and 300 MHz. AB - PURPOSE: To examine relationships between specific energy absorption rate (SAR) and temperature distributions in the human head during radio frequency energy deposition in MRI. MATERIALS AND METHODS: A multi-tissue numerical model of the head was developed that considered thermal conductivity, heat capacity, perfusion, heat of metabolism, electrical properties, and density. Calculations of SAR and the resulting temperature increase were performed for different coils at different frequencies. RESULTS: Because of tissue-dependent perfusion rates and thermal conduction, there is not a good overall spatial correlation between SAR and temperature increase. When a volume coil is driven to induce a head average SAR level of either 3.0 or 3.2 W/kg, it is unlikely that a significant temperature increase in the brain will occur due to its high rate of perfusion, although limits on SAR in any 1 g of tissue in the head may be exceeded. CONCLUSION: Attempts to ensure RF safety in MRI often rely on assumptions about local temperature from local SAR levels. The relationship between local SAR and local temperature is not, however, straightforward. In cases where high SAR levels are required due to pulse sequence demands, calculations of temperature may be preferable to calculations of SAR because of the more direct relationship between temperature and safety. PMID- 15112318 TI - Bromodomain containing 2 (Brd2) is expressed in distinct patterns during ovarian folliculogenesis independent of FSH or GDF9 action. AB - We previously observed high levels of Brd2 (also known as female sterile homeotic related gene-1, Fsrg1) expression in several hormonally responsive tissues, including the ovary. Here, we report distinct localization patterns of Brd2 transcripts throughout ovarian folliculogenesis in normal mice as well as in two strains of mice with aberrant folliculogenesis: mice with mutated growth differentiation factor 9 (Gdf9) and follicle stimulating hormone beta (Fshb) genes. The highest level of expression was seen in granulosa cells of growing follicles. Within the oocyte, three patterns of Brd2 RNA localization were observed: diffuse distribution in both the cytoplasm and nucleus, then intense nuclear expression, followed by an absence of Brd2 transcripts from the nucleus. The transition from intense nuclear localization to nuclear exclusion was found to correlate with oocyte maturation and meiotic competence, as determined by nuclear chromatin patterns. These same expression patterns were also seen in oocytes from Gdf9(-/-) and Fshb(-/-) mice. Thus, Brd2 expression appears to correlate with stages of oocyte maturation, independent of FSH or GDF9 action and the subsequent disruption in normal follicle development in these models. The distinct patterns of Brd2 localization within the adult ovary supports a role for Brd2 in mitotic and possibly meiotic cell cycle regulation. PMID- 15112319 TI - Role of basic helix-loop-helix (bHLH) and CREB transcription factors in the regulation of Sertoli cell androgen-binding protein expression. AB - Differentiation of Sertoli cells is marked by the presence of novel gene products such as transferrin and androgen-binding protein (ABP). Transcriptional regulation of Sertoli cell differentiation is, in part, controlled through the binding of specific transcription factors to response elements within these genes promoters. Transferrin gene expression has been shown to be regulated by the binding and interactions of basic helix-loop-helix (bHLH) and cAMP response element binding protein (CREB) to an E-box and cyclic AMP response element (CRE), respectively. Interaction between the bHLH and CREB is facilitated through subsequent binding of CREB-binding protein (CBP)/p300. The hypothesis tested in the current study is that ABP expression is regulated by a similar mechanism. The ABP promoter activation was analyzed through the use of transfection assays, site directed mutagenesis, and electromobility shift assays (EMSA). Transient transfections of rat Sertoli cells used a reporter construct containing the proximal 619 bp of the ABP promoter. Observations suggest that cAMP and follicle stimulating hormone (FSH) upregulate the expression of ABP. Mutational studies of the three E-boxes and the CRE of the 619-bp ABP promoter indicate that all of these elements are critical for stimulation of promoter activity. EMSA revealed a weak interaction between an E-box-2 and the CRE that are overlapping in the promoter. An artificial promoter that contains only an E-box and CRE was created to further test this hypothesis. The artificial promoter was stimulated by both FSH and cAMP. Experiments with mutants of the artificial promoter demonstrate that both response elements contribute to the optimal activation of the promoter construct. The overexpression of the bHLH inhibitor Id (i.e., inhibitor of differentiation) that binds bHLH proteins and eliminates DNA binding was found to suppress hormone activation of the ABP promoter. Combined observations of the ABP promoter and artificial promoter provide insight into a common mechanism for gene regulation in differentiated Sertoli cells involving a role for both the bHLH and CREB family of transcription factors. PMID- 15112320 TI - Cycle-dependent endometrial expression and hormonal regulation of the fibulin-1 gene. AB - Fibulin-1 is a secreted protein associated with elastic matrix fibres and basement membranes. It plays a role in stabilizing blood vessels and can also regulate cell motility and invasiveness. We studied the regulation of the fibulin 1 gene in the rat and human endometrium, an organ where cyclic tissue remodeling and angiogenesis take place. The rat fibulin-1C and -1D-specific DNA sequences were first identified and a comparison of the deduced amino acid sequence with the mouse and human counterparts showed a very strong conservation. The exon intron structure was also maintained. Primers were derived for RT-PCR analysis of fibulin-1 expression in rat endometrium. The highest levels of fibulin-1C and -1D transcripts were measured at metestrous and diestrous, and in early pregnancy at day 3 post-coitum. In vivo studies showed stimulation of endometrial fibulin-1D expression after estrogen application, an effect prevented by parallel treatment with progesterone. Analysis of human endometrial tissues indicated that the fibulin-1D transcript levels were higher during the mid-secretory phase than during the proliferative and early secretory phases. Cultured human endometrial stromal cells treated with progesterone responded with a dramatic increase of fibulin-1 protein expression. This was enhanced by parallel treatment with epidermal growth factor and prevented by application of the antiprogestin RU486. Altogether the results show a cycle-dependent regulation of endometrial fibulin-1 expression controlled by both progesterone and estrogen. Based on its implication in tissue remodeling and angiogenesis, fibulin-1 may play an important role in endometrial receptivity for embryo implantation. PMID- 15112321 TI - Global gene expression analysis comparing bovine blastocysts flushed on day 7 or produced in vitro. AB - In vitro produced (IVP) bovine embryos have darker cytoplasm, reduced buoyant density, fragile zonae pellucidae, chromosomal abnormalities, higher pregnancy failure rates, and altered gene expression compared to embryos produced in vivo. Characterization of early deviations in gene expression would enable us to better understand the biology of early embryo development and improve in vitro culture systems. Here we compared gene expression between Day 7 blastocysts generated in TCM199 with 5% FBS and Day 7 in vivo derived blastocysts and using suppression subtractive hybridization (SSH). Pools of 25 embryos for both driver and tester were used in the RNA extraction process. The subtracted products were cloned and subjected to differential hybridization screening analysis. cDNAs were isolated, single-pass sequenced, and subjected to BLAST search. Of 32 in vivo ESTs (expressed sequence tags) that provided sequence information, 30 matched homologous sequences in GenBank. Of 32 in vitro ESTs, 22 provided specific matches while the remaining ten represented novel transcripts. Two in vivo ESTs, galectin-1 and fibronectin, and one in vitro EST, filamin A, were further characterized using real-time quantitative PCR. To further examine the reproducibility of the SSH data, three different pools of embryos with each pool containing ten embryos produced from each of the following production systems, namely, in vivo, IVP in TCM199 with 5% FBS and CR1aa with 5% FBS were used for real-time reverse transcription-polymerase chain reaction (RT-PCR) confirmation studies. Significant increases in the expression level of galectin-1 and fibronectin were observed in the in vivo derived blastocysts compared to blastocysts produced in TCM199 with 5% FBS and CR1aa cultures. No significant difference in filamin A expression was found between blastocysts produced in vivo and those derived from either of the in vitro production systems. We conclude that these techniques are useful to characterize the transcriptome of the early preattachment embryo and observed deviations in mRNA expression may partially explain the differences in quality between in vivo and IVP embryos. PMID- 15112322 TI - Obtaining mice that carry human mitochondrial DNA transmitted to the progeny. AB - To study human diseases associated with mutations in mitochondrial DNA one needs an animal model in which the distribution of abnormal mtDNA and its impact on the phenotype might be followed. We isolated human mitochondria from HepG2 cell culture and microinjected them into murine zygotes, upon which those were transplanted to the pseudopregnant mice. PCR with species-specific primers allowed detecting human mtDNA in the tissues of 7-13-day embryos. No serious alterations in the development of transmitochondrial embryos were noticed. Among various organs/tissues of the 13-day embryos, human mtDNA was detected only in the heart, skeletal muscles, and stomach, which is in line with its uneven distribution among the blastomeres of an early mouse embryo that we described previously. In four recipient females, the microinjected zygotes were allowed to develop to term, the four neonate males of their joint litter were sacrificed, and in three of them human mtDNA was detected in the heart, skeletal muscles, stomach, brain, testes, and bladder. Six females of that joint litter were grown and mated to intact males. In the progeny (F1) of one of the females two mice were carrying human mtDNA in the heart, skeletal muscles, stomach, brain, lungs, uterus, ovaries, and kidneys. The study confirms the possibility to obtain transmitochondrial mice carrying human mtDNA that is transmitted to the animals of the next generation. Our results also indicate that among the organs to which human mtDNA is distributed some are more likely to receive it than others. PMID- 15112323 TI - Both blastomeres of the mouse 2-cell embryo contribute to the embryonic portion of the blastocyst. AB - To track the lineage of both blastomeres of 2-cell embryos during mouse preimplantation development, each cell was injected with dextran solutions conjugated with different fluorochromes. The fate of the progeny of the first two blastomeres was followed with confocal microscopy during cleavage and during the formation of the blastocyst. We observed that in most of cleaving embryos the cells derived from the two first blastomeres intermingled in both the trophectoderm and the inner cell mass (ICM) and did not form two discrete groups. We conclude that embryonic parts of blastocysts contain descendants of both blastomeres of 2-cell embryo. PMID- 15112324 TI - Differential effect of recipient cytoplasm for microtubule organization and preimplantation development in rat reconstituted embryos with two-cell embryonic cell nuclear transfer. AB - In the present study, we examined the developmental ability of enucleated zygotes, MII oocytes, and parthenogenetically activated oocytes at pronuclear stages (parthenogenetic PNs) as recipient cytoplasm for rat embryonic cell nuclear transfer. Enucleated zygotes as recipient cytoplasm receiving two-cell nuclei allowed development to blastocysts, whereas the development of embryos reconstituted with MII oocytes and parthenogenetic PNs was arrested at the two cell stage. Previous observations in rat two-cell embryos suggested that the distribution of microtubules is involved in two-cell arrest. Therefore, we also examined the distribution of microtubules using immunofluorescence. At the two cell stage after nuclear transfer into enucleated zygotes, microtubules were distributed homogeneously in the cytoplasm during interphase, and normal mitotic spindles were observed in cleaving embryos from the two- to four-cell stage. In contrast, embryos reconstituted with MII oocytes and parthenogenetic PNs showed aberrant microtubule organization. In enucleated zygotes, fibrous microtubules were distributed homogeneously in the cytoplasm. In contrast, dense microtubules were localized at the subcortical area in the cytoplasm and strong immunofluorescence intensity was observed at the plasma membrane, while very weak intensity was detected in the central part of enucleated MII oocytes. In enucleated parthenogenetic PNs, high-density and fibrous microtubules were distributed in the subcortical and central areas, respectively. Pre-enucleated parthenogenetic PNs also showed lower intensity of microtubule immunofluorescence in the central cytoplasm than zygotes. In conclusion, the results of the present study showed that zygote cytoplasm is better as recipient than MII oocyte and parthenogenetic PNs for rat two-cell embryonic cell nuclear transfer to develop beyond four-cell stage. Furthermore, microtubule organization is involved in the development of reconstituted embryos to overcome the two-cell arrest. PMID- 15112325 TI - Developmental changes in inhibin alpha and inhibin/activin betaA and betaB mRNA levels in the gonads during post-hatch prepubertal development of male and female chickens. AB - Dimeric inhibins and activins are barely detectable in the plasma during prepubertal development of male and female chickens. This may be misconstrued to indicate that the proteins are not produced in the gonads and have no functional significance during this period. Very few studies have actually determined the mRNA expression profile of the inhibin and activin subunits in the gonads prior to puberty in order to establish their secretion at the local level and postulate potential roles for the inhibin and activins at this developmental stage. In this study, the expression of the mRNA for the alpha-, betaA-, and betaB-subunits was determined in the ovary and testis of chickens during prepubertal development. Gene expression was determined at 3, 5, 6, 8, 10, 12, 16, and 18 weeks of age by RT-PCR. Messenger RNA level was quantified by competitive RT-PCR at 3, 6, 12, and 18 weeks of age in order to detect any changes with development, suggest potential relationship to the profile of dimeric inhibins and activins reported previously and to suggest potential paracrine and endocrine roles for them. The results show that all the inhibin/activin subunit mRNAs are expressed in the testis of the chicken throughout the period of prepubertal development up to 18 weeks of age. However, in the ovary, only the betaA- and betaB-subunits were detected at all ages whereas the alpha-subunit mRNA could only be detected just before puberty. Quantification of the mRNA levels showed variation of each subunit with age. These temporal changes suggest relationship with paracrine functional role in the ovary or the testis. Quantitative changes in expression levels also suggests that there may be some relationship between mRNA levels and the type and amount of dimeric inhibins and activins produced at any developmental stage. There are major differences between the male and female gonads in the timing of the expression of different subunits. In conclusion, the expression of the mRNA subunits in the testis and ovary suggests that inhibins and activins are being produced but may be principally involved in autocrine/paracrine function within the gonads. PMID- 15112326 TI - Nucleolar ultrastructure and protein allocation in in vitro produced porcine embryos. AB - The nucleolus formation was studied as an indirect marker of the ribosomal RNA (rRNA) genes activation in porcine embryos following oocyte maturation, fertilization, and culture in vitro. Nucleologenesis was assessed by transmission electron microscopy (TEM), light microscopical autoradiography following 20 min of 3H-uridine incubation, and immunocytochemical localization of key nucleolar proteins involved in rRNA transcription (upstream binding factor (UBF), topoisomerase I, and RNA polymerase I) and processing (fibrillarin, nucleophosmin, nucleolin) by confocal laser scanning microscopy. During the first four post-fertilization cell cycles, TEM revealed spherical nucleolus precursor bodies (NPBs), consisting of densely packed fibrils, as the most prominent intra nuclear entities of the blastomeres. Fibrillo-granular nucleoli were observed in some blastomeres in a single embryo during the 5th cell cycle, i.e., the tentative 16-cell stage, where formation of fibrillar centres (FC), a dense fibrillar component, and a granular component on the surface of the NPBs was seen. In this embryo, autoradiographic labeling was detected over the nucleoplasm and in particular over the nucleoli. Fibrillarin was immunocytochemically localized in the presumptive NPBs of the pronuclei. This protein was again localized to the presumptive NPBs together with nucleolin from late during the 3rd cell cycle, i.e., the four-cell stage in some embryos. UBF, RNA polymerase I, and nucleophosmin were localized to the presumptive NPBs in a proportion of the embryos at the 4th cell cycle, i.e., the tentative eight-cell stage and onwards. Toposiomerase I was not localized to intra-nuclear entities even during the 5th post-fertilization cell cycle. Moreover, a considerable proportion of the blastomere nuclei apparently did not show localization of other nucleolar proteins. In conclusion, porcine embryos produced in vitro display a substantial delay in or even lack of the development of functional nucleoli. PMID- 15112327 TI - Hepatocyte growth factor promotes cell proliferation and inhibits progesterone secretion via PKA and MAPK pathways in a human granulosa cell line. AB - Hepatocyte growth factor (HGF) is a mesenchymal-derived paracrine factor that acts through a c-met receptor. The activated c-met receptor recruits various signal proteins. We used a steroidogenic human granulosa-like tumor cell line (KGN cells) to analyze the biological function of HGF in human ovary cells. First, we designed a method to analyze local production and action of HGF in the human ovary. Although c-met mRNA is expressed in KGN cells, granulosa lutein, theca, and ovarian stroma cells, we observed HGF mRNA only in theca and stroma cells. Adding HGF to the medium enhanced mitogenic activity in KGN cells. We next examined the activation of intracellular signal transduction molecules induced by HGF in KGN cells. Here, we showed that HGF activated the distinct phosphorylation of Raf-1, MEK1/2, and ERK1/2, but did not induce phosphorylation of Akt. HGF enhanced the phosphorylation of Elk-1 and c-Jun as nuclear transcription factors. U0126, a MEK1/2 inhibitor, completely abrogated the phosphorylation of ERK1/2 and the cell proliferation in response to HGF. In contrast, H-89, a protein kinase A inhibitor, further enhanced the HGF-induced phosphorylation of ERK1/2 and cell proliferation. In addition, we revealed that HGF suppressed progesterone synthesis in KGN cells. Adding HGF suppressed the forskolin-induced steroidogenic acute regulatory protein (StAR) expression, which is a key regulator in progesterone synthesis. Crosstalk signals between PKA and the mitogen-activated protein kinase (MAPK) pathway were mutually inhibitory. These results demonstrated for the first time that theca cell-derived HGF may be capable of stimulating the proliferation of granulosa cells and suppressing progesterone synthesis via an activating MAPK pathway. PMID- 15112328 TI - Histamine enhances cytotrophoblast invasion by inducing intracellular calcium transients through the histamine type-1 receptor. AB - Blastocyst implantation and placentation require molecular and cellular interactions between the uterine endometrium and blastocyst trophectoderm. Previous studies showed that histamine produced in the mouse uterine luminal epithelium interacts with trophoblast histamine type-2 receptors (H2) to initiate blastocyst implantation. However, it is unknown whether similar histamine activity is operative in humans. Using a human cell line (HTR-8/SVneo) derived from first-trimester cytotrophoblasts that expresses both histamine type-1 receptor (H1) and H2, we found that histamine promotes cytotrophoblast invasiveness specifically through activation of H1. Stimulation of H1 in human cytotrophoblasts by histamine induced intracellular Ca2+ (Ca(2+)i) transients by activating phospholipase C and the inositol trisphosphate pathway. The enhanced invasion induced by histamine was blocked by pretreatment with H1 antagonist or by chelation of Ca(2+)i. These findings suggest possible differences between rodents and humans in histamine signaling to the trophoblast. PMID- 15112329 TI - Molecular characterization of the 32 kDa boar sperm protease. AB - The purified 32 kDa boar sperm protease, known as the 32 kDa sperminogen, was identified by reverse transcription polymerase chain reaction (RT-PCR) and Northern blot analysis following its partial peptide sequencing. The 32 kDa boar sperm protease was purified from the acid extracts of boar spermatozoa and subjected to CNBr-digestion. The most prominently digested 30 kDa product was purified by HPLC and its peptide sequence was analyzed. NCBI Blast search of the analyzed 21 amino acid sequence revealed that the sequence matched 91% with that of proacrosin. DNA primer was deduced from the analyzed peptide sequence and the 32 kDa protease was further identified by RT-PCR. Upon RT-PCR, 1 Kbp DNA fragment was amplified, which is the expected length if the product was amplified from the proacrosin mRNA, implying that there is no separate mRNA for the 32 kDa sperminogen. To confirm these results, Northern blot analysis was performed. Four DNA probes generated from the exons of proacrosin genomic DNA sequence all detected a single species of mRNA, suggesting that there is no separate mRNA for the 32 kDa sperminogen which might be produced either from the potential separate 32 kDa sperminogen gene or by differential splicing from proacrosin mRNA. These results strongly suggest that the 32 kDa protease is part of the proacrosin/acrosin system, and that the 32 kDa sperminogen might be formed from post-translational processing of proacrosin. PMID- 15112330 TI - Characterization of the major egg glycolipoproteins from the perivitellin fluid of the apple snail Pomacea canaliculata. AB - Ovorubin and PV2 are the major lipoglycocarotenoproteins present in the perivitellus of the freshwater snail eggs of Pomacea canaliculata, a rapidly expanding rice field pest. We have previously characterized these two particles regarding their lipid and protein compositions, their synthesis and tissular distribution, and their contributions of energy and structural precursors for the developing embryo. In the present study, we have characterized the glycosidic moieties associated to these perivitellines. Both proteins were isolated from egg homogenates by ultracentrifugation, and high performance liquid chromatography (HPLC) using anionic exchange and size exclusion columns. Total carbohydrates accounted for 17.8% and 2.5% (w/w) of the apparent molecular mass of ovorubin and PV2, respectively. Analysis by size exclusion chromatography showed that the amount of O-linked oligosaccharides is higher than that of the N-linked species (59% and 67% w/w of total carbohydrates of ovorubin and PV2, respectively). Glycosylation patterns were determined by a set of biotinilated lectins onto blotted purified proteins. Lectin affinities confirmed the presence of aspargine linked carbohydrates, probably of hybrid and high mannose types. Jacaline affinity suggested the presence of O-linked residues derived from the T-antigen. Total carbohydrate composition determined by gas liquid chromatography (GLC) showed that mannose was the major monosaccharide in both perivitellins followed by GlcNAc and Gal in ovorubin, and Gal and GlcNAc in PV2. Only one fatty acid (22:1 n-9) accounted for 46% and 56% of the fatty acids present in ovorubin and PV2, respectively. Carbohydrate role on these reserve proteins during embryogenesis of the apple snail is discussed. PMID- 15112331 TI - Lipid diffusion in sperm plasma membranes exposed to peroxidative injury from oxygen free radicals. AB - Unsaturated lipids in sperm plasma membranes are very susceptible to peroxidation when exposed to reactive oxygen species (ROS). In this investigation we have incubated ram spermatozoa in the presence of two ROS generating systems, ascorbate/FeSO4 and potassium peroxychromate (K3CrO8), and examined their effects on membrane fluidity by measuring fluorescence recovery after photobleaching (FRAP) of a lipid reporter probe 5-(N-octadecanoyl)-aminofluorescein (ODAF). Peroxidation was monitored by malonaldehyde formation and changes in fluorescence emission of 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-diaza-s indacene-3-undecanoic acid (C11-BODIPY(581/591)). Ascorbate/FeSO4-induced peroxidation was inhibited by Vitamin E, butylated hydroxyanisole (BHA), 1,4 diazobicyclo(2,2,2)octane (DABCO), and to a lesser extent by ethanol. Added superoxide dismutase (SOD), gluthathione peroxidase (GPX), and catalase were ineffective scavengers. K3CrO8 induced very rapid peroxidation that could be delayed, but not prevented, by Vitamin E, BHT, DABCO, ethanol, and mannitol; once again SOD, GPX, and catalase were ineffective scavengers. Neither peroxidation with ascorbate/FeSO4 nor K3CrO8, or added H2O2 or malonaldehyde perturbed ODAF diffusion in any region of the sperm plasma membrane. Vitamin E tended to enhance diffusion rates. Exogenous cumene hydroperoxide, however, reduced ODAF diffusion to low levels on the sperm head. These results suggest that the adverse effects of ROS on spermatozoa are more likely to be caused by direct oxidation of proteins and membrane permeabilisation than disturbance of lipid fluidity. PMID- 15112332 TI - Hamster contraception associated protein 1 (CAP1). AB - Based on cDNA and amino acid sequence, we demonstrate that hamster contraception associated protein 1 (CAP1) protein (an homolog of DJ-1 in mouse, CAP1/SP22/RS in rat and DJ-1/RS in human) is conserved during evolution. Through solubilization studies, it was demonstrated that hamster CAP1 has a peripheral membrane localization. SDS-PAGE analysis revealed that the migration pattern for hamster CAP1 compared to the other rodent counterparts, rat and mouse was different; indicating species-specific differences in the protein (possibly due to post translational modifications). This protein also shows a ubiquitous presence in both somatic and germ tissues, and has been localized to the sperm tail. It was noticed that hamster CAP1 was lost from the mid piece of spermatozoa during capacitation. Interestingly, following in vitro treatment with ornidazole, CAP1 was lost from the spermatozoa and immunofluorescence studies showed that the major loss was from the mid piece of the spermatozoa. Another interesting feature highlighted about hamster CAP1 is its tendency to exist in two pI isoforms. Summarily, hamster CAP1 appears to exhibit species-specific differences compared to its rodent counterparts with respect to its unique peripheral localization, its size, two pI isoforms, and fate during capacitation, which may have implications in its functions. PMID- 15112334 TI - Wireless capsule video endoscopy: three years of experience. AB - AIM: To review and summarize the current literature regarding M2A wireless capsule endoscopy. METHODS: Peer reviewed publications regarding the use of capsule endoscopy as well as our personal experience were reviewed. RESULTS: Review of the literature clearly showed that capsule endoscopy was superior to enteroscopy, small bowel follow through and computerized tomography in patients with obscure gastrointestinal bleeding, iron deficiency anemia, or suspected Crohn's disease. It was very sensitive for the diagnosis of small bowel tumors and for surveillance of small bowel pathology in patients with Gardner syndrome or familial adenomatous polyposis syndrome. Its role in celiac disease and in patients with known Crohn's disease was currently being investigated. CONCLUSION: Capsule video endoscopy is a superior and more sensitive diagnostic tool than barium follow through, enteroscopy and entero-CT in establishing the diagnosis of many small bowel pathologies. PMID- 15112335 TI - GSTT1, GSTM1 and CYP2E1 genetic polymorphisms in gastric cancer and chronic gastritis in a Brazilian population. AB - AIM: To test the hypothesis that, in the Southeastern Brazilian population, the GSTT1, GSTM1 and CYP2E1 polymorphisms and putative risk factors are associated with an increased risk for gastric cancer. METHODS: We conducted a study on 100 cases of gastric cancer (GC), 100 cases of chronic gastritis (CG), and 150 controls (C). Deletion of the GSTT1 and GSTM1 genes was assessed by multiplex PCR. CYP2E1/PstI genotyping was performed using a PCR-RFLP assay. RESULTS: No relationship between GSTT1/GSTM1 deletion and the c1/c2 genotype of CYP2E1 was observed among the three groups. However, a significant difference between CG and C was observed, due to a greater number of GSTT1/GSTM1 positive genotypes in the CG group. The GSTT1 null genotype occurred more frequently in Negroid subjects, and the GSTM1 null genotype in Caucasians, while the GSTM1 positive genotype was observed mainly in individuals with chronic gastritis infected with H pylori. CONCLUSION: Our findings indicate that there is no obvious relationship between the GSTT1, GSTM1 and CYP2E1 polymorphisms and gastric cancer. PMID- 15112336 TI - Role of nucleostemin in growth regulation of gastric cancer, liver cancer and other malignancies. AB - AIM: To examine the role of nucleostemin in the growth regulation of gastric cancer, liver cancer and other cancers. METHODS: RT-PCR was used to clone the fragment of nucleostemin cDNA from HEK 293 cells. Eighteen kinds of malignant tumor tissues including gastric adenocarcinoma and liver cancer tissues, 3 kinds of benign tumor tissues, 3 kinds of benign hyperplastic tissues and normal tissues were employed to examine nucleostemin gene expression by RT-PCR, Slot blot, Northern blot and in situ hybridization. RESULTS: We successfully cloned a 570 bp fragment of nucleostemin-cDNA from HEK-293 cells. All detected malignant tumor tissues, benign tumor tissues, and benign hyperplastic tissues had high levels of nucleostemin expression. Nucleostemin was also expressed in human placenta tissue at a high level. In terminally differentiated normal human adult kidney and mammary gland tissues, no nucleostemin expression could be detected. CONCLUSION: Nucleostemin can help regulate the proliferation of both cancer cells and stem cells. It might play an important role in the growth regulation of gastric cancer, liver cancer and other cancers. PMID- 15112337 TI - Significance of vascular endothelial growth factor expression and its correlation with inducible nitric oxide synthase in gastric cancer. AB - AIM: To investigate the clinical significance of the expression of VEGF165mRNA and the correlation with vascular endothelial growth factor (VEGF) protein and inducible nitric oxide synthase (iNO) in human gastric cancer. METHODS: We tested VEGF165mRNA expression in 31 cases of resected gastric cancer specimens and normal paired gastric mucosae by RT-PCR. Total RNA was extracted with TRIzol reagents, transcribed into cDNA with oligo (dT15) priming, inner controlled with beta-actin expression and agarose gel isolated after PCR. VEGF expression was quantitated with IS1000 imaging system. Meanwhile we also examined expression levels of VEGF protein and iNOS in 85 cases of gastric cancer. All paraffin embedded samples were immunohistochemically stained by streptavidin -peroxidase method (SP). RESULTS: The mean expression of VEGF165mRNA in gastric cancer was 1.125+/-0.356, significantly higher than that of normal paired mucosae, which was 0.760+/-0.278. The data indicated that the expression level of VEGF165mRNA was well related to lymph node metastasis and TNM stages of UICC. The expression levels in patients with lymph node metastasis and without lymph node metastasis were 1.219+/-0.377 and 0.927+/-0.205 respectively (P<0.05). The expression in stages I, II, III, IV was 0.934+/-0.194, 1.262+/-0.386 respectively (P<0.01). Further analysis showed the lymph node metastasis rate in the group with over expression of VEGF was higher than that in the group with low expression of VEGF (83.3% vs 46.2%), and the ratio of stage III+IV in the group with over-expression of VEGF was also higher than that in the group with low expression with VEGF (77.8% vs 33.8%) (P<0.05). The positive rates of expression of VEGF protein and iNOS in 85 cases of gastric cancer were 75.4% and 58.8% respectively, and 50.1% of the patients showed positive staining both for iNOS and VEGF, the correlation with the two factors was significant (P=0.018). But more intensive analysis showed the immunoreactive grades of VEGF were not associated with that of iNOS. CONCLUSIONS: The expression of VEGF165mRNA is well related with lymph node metastasis and TNM stages of UICC in gastric cancer, and is concerned with the invasiveness and metastasis of gastric cancer. The relationship can be observed between the expression of VEGF and iNOS in gastric cancer. PMID- 15112338 TI - Association of tumor necrosis factor genetic polymorphism with chronic atrophic gastritis and gastric adenocarcinoma in Chinese Han population. AB - AIM: To investigate the association of TNF polymorphisms with chronic atrophic gastritis (CAG) and gastric adenocarcinoma in Chinese Han patients. METHODS: The TNFa-e 5 microsatellites and 3 RFLP sites were typed using PCR technique, followed by high-voltage denaturing PAGE with silver staining and restriction enzyme digestion respectively in specimens from 53 patients with CAG and 56 patients with gastric adenocarcinoma and 164 healthy controls. The PCR products were cloned and sequenced. RESULTS: The frequency of TNF-beta Ncol*1/2 genotype was higher in patients with chronic atrophic gastritis than in healthy controls, but no significant difference was observed (60.38% vs 46.34%, P=0.076). The frequency of TNa10 allele was significantly higher in patients with chronic atrophic gastritis than in healthy controls (19.81% vs 11.89%, P=0.04). However, it did not relate to age, gender, atrophic degree or intestinal metaplasia in patients with chronic atrophic gastritis. The frequency of TNF-beta Ncol*1/2 and d2/d6 genotypes were significantly higher in patients with gastric adenocarcinoma than in healthy individuals (P>0.05). However, TNF-beta Ncol*1/2 and d2/d6 genotypes did not relate to age, gender, grade of differentiation and clinicopathologic stage in patients with gastric adenocarcinoma. The frequency of TNFa6b5c1 haplotype homozygote was significantly lower in patients with gastric adenocarcinoma than in healthy controls (1.79% vs 15.85%, P=0.006). CONCLUSION: TNFa10 allele may be a risk factor for chronic atrophic gastritis. TNF-beta Ncol*1/2 and d2/d6 genotypes are associated with the susceptibility to gastric adenocarcinoma, whereas TNFa6b5c1 haplotype homozygote may contribute to the resistance against gastric adenocarcinoma. PMID- 15112339 TI - Significance of effector protease receptor-1 expression and its relationship with proliferation and apoptotic index in patients with primary advanced gastric adenocarcinoma. AB - AIM: To investigate the expression of effector protease receptor-1 (EPR-1), proliferative index ki-67 and apoptosis index in patients with primary advanced gastric adenocarcinoma and to clarify the significance of EPR-1 expression and its correlationship with the proliferation and apoptosis indexes. METHODS: Using immunohistochemical staining and terminal deoxynucleotidyl transferase mediated nick end labelling (TUNEL) technique, we determined the expression of EPR-1, proliferative index (Ki-67) and apoptotic index (AI) in 120 paraffin-embedded specimens of primary advanced gastric adenocarcinoma as well as lymph node metastasis and adjacent normal tissues. RESULTS: EPR-1 expression was distributed in the cytoplasm of normal gastric mycoderma, carcinoma cells and smooth muscle cells. The positive rate of EPR-1 expression in the primary gastric adenocarcinomas, invasion tumor node and lymph node metastasis was 65.83%, 55.29% and 68%, respectively. While the positive rate in normal gastric mycoderma and smooth muscle cells was 46.7% and 53.3%, respectively. The average positive rate of ki-67 in EPR-1-positive tumors was 7.00% which was significantly lower than that of 8.53% in EPR-1-negative tumors, but the average AI in EPR-1-positive tumors was 1.25%, which was significantly higher than that of 1.00% observed in EPR-1-negative tumors. On the other hand, the average positive labeling index for Ki-67 (ki-67) in EPR-1-positive lymph node metastasis was 7.65%, which was significantly lower than that of 9.44% observed in EPR-1-negative lymph node metastasis. However, the average AI in EPR-1-positive lymph node metastasis tumors was 0.99%, which was significantly higher than that of 0.67% observed in EPR-1-negative lymph node metastasis. CONCLUSION: The frequency of EPR-1 expression was significantly higher in primary gastric adenocarcinoma and in its lymph node metastasis than that in normal gastric mucosa. Expression of EPR-1 was significantly correlated with tumor histological subtypes and tumor differentiation. Weighted EPR-1 Score is positively correlated with apoptosis index, but is negatively related with proliferative index. Thus, Weighted EPR-1 Score and EPR-1 expression in gastric adenocarcinoma cells maybe a potential marker in clinical setting. PMID- 15112340 TI - 1alpha, 25-dihydroxyvitamin D3 prevents DNA damage and restores antioxidant enzymes in rat hepatocarcinogenesis induced by diethylnitrosamine and promoted by phenobarbital. AB - AIM: To investigate the chemopreventive effects of 1alpha, 25-dihydroxyvitamin D(3) in diethylnitrosamine induced, phenobarbital promoted rat hepatocarcinogenesis. METHODS: The rats were randomly divided into 6 different groups (A-F). Groups A, C and E rats received a single intraperitoneal (i.p.) injection of diethylnitrosamine (DEN) at a dose of 200 mg/kg body mass in 9 g/L NaCl solution at 4 wk of age, while group B served as normal vehicle control received normal saline once. After a brief recovery of 2 wk, all the DEN treated rats were given phenobarbital (PB) at 0.5 g/L daily in the basal diet till wk 20. Group A was DEN control. Treatment of 1alpha, 25-(OH)2D3 in group C was started 4 wk prior to DEN injection and continued thereafter till wk 20 at a dose of 0.3 microg/100 microL propylene glycol per one single dose (os) twice a week. Group E received the treatment of 1alpha, 25-(OH)2D3 at the same dose mentioned as above for 15 wk. The rats in group D and F received 1alpha, 25-(OH)2D3 alone as in group C without DEN injection. RESULTS: The comet assay showed statistically higher mean values for length to width ratios (L: W) of DNA mass and tailed cells (P<0.01; P<0.01 respectively) in DEN treated rats as compared to their normal controls. Continuous supplementation of 1alpha, 25-dihydroxyvitaminD2 showed a significant (P<0.01) decrease in L:W ratio of DNA mass tailed cells. Furthermore, 1alpha, 25-(OH)2D3 supplementations elevated the super oxide dismutase (SOD) activity, hepatic malondialdehyde (MDA) level, reduced glutathione (GSH) and glutathione S-transferase (GST) activity (P<0.01, P<0.05, P<0.05 and P<0.05 respectively). As an endpoint marker histological changes were observed to establish the chemopreventive effects of 1alpha, 25-dihydroxyvitaminD2. CONCLUSION: Supplementations of 1alpha, 25-(OH)2D3 has a marked protection against hepatic nodulogenesis, antioxidant enzymes and DNA damages in DEN induced rat hepatocarcinogenesis promoted by phenobarbital. PMID- 15112341 TI - Association of low p16INK4a and p15INK4b mRNAs expression with their CpG islands methylation with human hepatocellular carcinogenesis. AB - AIM: To study the significance of p16 and p15 transcription suppression with hypermethylation of their genes' 5'CpG islands during human hepatocellular carcinogenesis. METHODS: The mRNA expression levels of p16 and p15 genes were evaluated in cancerous, para-cancerous and non-cancerous tissues of 20 HCC, 3 normal liver tissues from 3 accidentally died healthy adults using semi quantitatively Northern blot. The methylation status was also assessed with methylation specific PCR. RESULTS: p16 mRNA expression level was decreased in the cancerous tissues in 60% (12/20) of HCC patients, of which 2 cases had no p16 mRNA detected, 5 cases (25%) displayed variation in the order of cancerous0.05). There was no statistical significance in the amino acid uptake rate according to the Dukes stage, though it was higher in patients with Dukes stage C or D than that with Dukes stage B (P>0.05). CONCLUSION: Abnormal synthetic metabolism of colon cancer may contribute to its higher amino acid uptake rate than that of normal colon. PMID- 15112346 TI - Construction of a metastasis-associated gene subtracted cDNA library of human colorectal carcinoma by suppression subtraction hybridization. AB - AIM: To construct a differentially-expressed gene subtracted cDNA library from two colorectal carcinoma (CRC) cell lines with different metastatic phenotypes by suppression subtractive hybridization. METHODS: Two cell lines of human CRC from the same patient were used. SW620 cell line showing highly metastatic potential was regarded as tester in the forward subtractive hybridization, while SW480 cell line with lowly metastatic potential was treated as tester in the reverse hybridization. Suppression subtractive hybridization (SSH) was employed to obtain cDNA fragments of differentially expressed genes for the metastasis of CRC. These fragments were ligated with T vectors, screened through the blue-white screening system to establish cDNA library. RESULTS: After the blue-white screening, 235 white clones were picked out from the positive-going hybridization and 232 from the reverse. PCR results showed that 200-700 bp inserts were seen in 98% and 91% clones from the forward and reverse hybridizations, respectively. CONCLUSIONS: A subtractive cDNA library of differentially expressed genes specific for metastasis of CRC can be constructed with SSH and T/A cloning techniques. PMID- 15112347 TI - Helicobacter pylori L-form and patients with chronic gastritis. AB - AIM: To study the relationship between infection with Helicobacter pylori L-forms and chronic gastritis and its association with possible changes of cellular immune function. METHODS: Gastric mucosal biopsies were taken from 428 patients with chronic gastritis to detect H pylori L-form by Gram staining and immunohistochemistry staining. Peripheral venous blood samples of patients were taken to detect the percentage of CD3+, CD4+ and CD8+ by the biotin-streptavidin (BSA) assay and the levels of IL-2, IL-6 and IL-8 by ELISA. RESULTS: The rate of infection with H pylori L-forms was 48.83% (209/428). The rate was 50.47% (216/428) and 52.80% (226/428), respectively, as detected by immunohistochemistry staining and Gram staining (P>0.05). The rate of H pylori L-forms in males and females was 57.8% (136/235) and 37.28% (73/193), respectively, (chi2=17.05, P<0.01). Furthermore, the rate increased with age, with the rate being significantly greater in patients > or =40 years old than in those <40 years old (P<0.01). The percentage of CD3+, CD4+, CD8+, the ratio of CD4+/CD8+, and the levels of IL-2, IL-6, IL-8 in H pylori-positive patients were 47.58+/-4.44%, 25.51+/-4.74%, 22.77+/-7.46%, 1.44+/-0.51%, 1.56+/-0.47 mg/L, 103.62+/-5.85 ng/L, and 109.79+/-7.18 ng/L, respectively. Compared with H pylori-negative patients, the percentage of CD3+, CD4+ and the ratio of CD4+/CD8+ and the IL-2 level decreased, but the levels of IL-6, IL-8 increased (P<0.001-P<0.01). Moreover, the percentage of CD3+, CD4+, CD8+, the ratio of CD4+/CD8+, and the levels of IL-2, IL-6 and IL-8 in patients infected with both H pylori L-forms and vegetative forms were 46.67+/-5.21%, 30.75+/-4.89%, 22.15+/-6.45%, 1.32+/-0.47%, 1.16+/-0.38 mg/L, 116.45+/-5.44 ng/L, and 118.64+/-6.24 ng/L, respectively. Compared with patients infected with only vegetative forms, the percentage of CD4+, the ratio of CD4+/CD8+and IL-2 level decreased, but the levels of IL-6 and IL-8 increased (P<0.001 or P<0.01). CONCLUSION: L-form variation often occurs in patients with chronic gastritis and is commonly found in male patients and associates with ages. The L-form variation may be an important factor causing disorder of cellular immune function in the patients with H pylori-induced chronic gastritis. PMID- 15112348 TI - C-kit gene mutation in human gastrointestinal stromal tumors. AB - AIM: To investigate the significance of c-kit gene mutation in gastrointestinal stromal tumors (GIST). METHODS: Fifty two cases of GIST and 28 cases of other tumors were examined. DNA samples were extracted from paraffin sections and fresh blocks. Exons 11, 9 and 13 of the c-kit gene were amplified by PCR and sequenced. RESULTS: Mutations of exon 11 were found in 14 of 25 malignant GISTs (56%), mutations of exon 11 of the c-kit gene were revealed in 2 of 19 borderline GISTs (10.5%), and no mutation was found in benign tumors. The mutation rate showed significant difference (chi2=14.39, P<0.01) between malignant and benign GISTs. Most of mutations consisted of the in-frame deletion or replication from 3 to 48 bp in heterozygous and homozygous fashions, None of the mutations disrupted the downstream reading frame of the gene. Point mutations and frame deletions were most frequently observed at codons 550-560, but duplications were most concentrated at codons 570-585. No mutations of exons 9 and 13 were revealed in GISTs, Neither c-kit gene expression nor gene mutations were found in 3 leiomyomas, 8 leiomyosarcomas, 2 schwannomas, 2 malignant peripheral nerve sheath tumors, 2 intra-abdominal fibromatoses, 2 malignant fibrous histiocytomas and 9 adenocarcinomas. CONCLUSION: C-kit gene mutations occur preferentially in malignant GISTs and might be a clinically useful adjunct marker in the evaluation of GISTs and can help to differentiate GISTs from other mesenchymal tumors of gastrointestinal tract, such as smooth muscle tumors, schwannomas, etc. PMID- 15112349 TI - Suppressive effects of 17beta-estradiol on hepatic fibrosis in CCl4-induced rat model. AB - AIM: To investigate the pathway via which 17beta-estradiol (beta-Est) exerts suppressive effects on rat hepatic fibrosis. METHODS: In vivo study was done in CCl4-induced female hepatofibrotic rats. Fibrosis-suppressive effect of beta-Est (20 microg/kg/d) was evaluated in intact and ovariectomized rat models. Six weeks after the treatment, all the rats were sacrificed and specimens of serum or liver tissue were collected for the studies. Serum liver enzymes, fibrosis markers and estradiol levels were determined by standard enzymatic methods, ELISA and RIA, respectively. Degrees of fibrosis and areas of hepatic stellate cells (HSCs) positive for alpha-smooth muscle actin (alpha-SMA) in the liver were determined by van Gieson (VG) stain and immunohistochemistry. In vitro studies, HSCs were isolated by a combination of pronase-collagenase perfusion and density gradient centrifugation. First-passage HSCs were randomly divided into 10 groups, and different concentrations of beta-Est, 2-hydroxyestradiol (2OHE) or 2 methoxyestradiol (2MeOE) were separately added to the cell groups. After incubation for 72 h, the degree of cell proliferation, collagen production, alpha SMA or estrogen receptor (ER) expression was determined by MTT assay, ELISA and immunohistochemistry, respectively. RESULTS: Beta-Est treatment reduced aspartate aminotransferase (AST), alanine aminotransferase (ALT), hyaluronic acid (HA) and type IV collagen (C IV) in sera, suppressed hepatic collagen content, decreased the areas of HSCs positive for alpha-SMA significantly in both intact and ovariectomized female hepatofibrotic rats. There was a negative correlation between the percentage of fibrotic area of liver tissue and the serum estradiol level; the calculated correlation coefficient was -0.57 (P<0.01). Beta-Est and its metabolites concentration-dependently (10(-9) mol/L-10(-7) mol/L) inhibited HSC proliferation and collagen synthesis. At the concentration of 10(-7) mol/L, they could inhibit alpha-SMA expression. The order of potency was 2MeOE>2OHE>beta Est. CONCLUSION: Beta-Est may suppress hepatic fibrosis probably via its biologically active metabolites. PMID- 15112350 TI - Hyperammonemia, brain edema and blood-brain barrier alterations in prehepatic portal hypertensive rats and paracetamol intoxication. AB - AIM: To study the blood-brain barrier integrity, brain edema, animal behavior and ammonia plasma levels in prehepatic portal hypertensive rats with and without acute liver intoxication. METHODS: Adults male Wistar rats were divided into four groups. Group I: sham operation; II: Prehepatic portal hypertension, produced by partial portal vein ligation; III: Acetaminophen intoxication and IV: Prehepatic portal hypertension plus acetaminophen. Acetaminophen was administered to produce acute hepatic injury. Portal pressure, liver serum enzymes and ammonia plasma levels were determined. Brain cortex water content was registered and trypan blue was utilized to study blood brain barrier integrity. Reflexes and behavioral tests were recorded. RESULTS: Portal hypertension was significantly elevated in groups II and IV. Liver enzymes and ammonia plasma levels were increased in groups II, III and IV. Prehepatic portal hypertension (group II), acetaminophen intoxication (group III) and both (group IV) had changes in the blood brain barrier integrity (trypan blue) and hyperammonemia. Cortical edema was present in rats with acute hepatic injury in groups III and IV. Behavioral test (rota rod) was altered in group IV. CONCLUSION: These results suggest the possibility of another pathway for cortical edema production because blood brain barrier was altered (vasogenic) and hyperammonemia was registered (cytotoxic). Group IV, with behavioral altered test, can be considered as a model for study at an early stage of portal-systemic encephalopathy. PMID- 15112351 TI - Modulation of Kupffer cells on hepatic drug metabolism. AB - AIM: To observe the effects of Kupffer cells on hepatic drug metabolic enzymes. METHODS: Kunming mice were i.p. injected with GdCl310, 20, 40 mg/kg to decrease the number and block the function of kupffer cells selectively. The contents of drug metabolic enzymes, cytochrome P450, NADPH-cytochrom C redutase (NADPH-C), aniline hydroxylase (ANH), aminopyrine N-demethylase (AMD), erythromycin N demethylase (EMD), and glutathione s-transferase (mGST) in hepatic microsome and S9-GSTpi, S9-GST in supernatant of 9 000 g were accessed 1 d after the injection. The time course of alteration of drug metabolic enzymes was observed on d 1, 3, and 6 treated with a single dose GdCl3. Mice were treated with Angelica sinensis polysaccharides (ASP) of 30, 60, 120 mg/kg, i.g., qd x 6 d, respectively and the same assays were performed. RESULTS: P450 content and NADPH-C, ANH, AMD, and EMD activities were obviously reduced 1 d after Kupffer cell blockade. However, mGST and S9-GST activities were significantly increased. But no relationship was observed between GdCl3 dosage and enzyme activities. With single dose GdCl3 treatment, P450 content, NADPH-C, and ANH activities were further decreased following Kupffer cell blockade lasted for 6 d, by 35.7%, 50.3%, 36.5% after 3 d, and 57.9%, 57.9%, 63.2% after 6 d, respectively. On the contrary, AMD, EMD, mGST, and S9-GST activities were raised by 36.5%, 71.9%, 23.1%, 35.7% after 3 d, and 155%, 182%, 21.5%, 33.7% after 6 d, respectively. Furthermore, the activities of drug metabolic enzymes were markedly increased after 30 mg/kg ASP treatment, and decreased significantly after 120 mg/kg ASP treatment. No change in activity of S9-GSTpi was observed in the present study. CONCLUSION: Kupffer cells play an important role in the modulation of drug metabolic enzymes. The changes of drug metabolic enzyme activities depend on the time of kupffer cell blockade and on the degree of Kupffer cells activated. A low concentration of ASP increases the activities of drug metabolic enzymes, but a high concentration of ASP decreases the activities of drug metabolic enzymes. PMID- 15112352 TI - Peroxisome proliferator activated receptor-gamma in pathogenesis of experimental fatty liver disease. AB - AIM: To study the expression of peroxisome proliferator activated receptor-gamma (PPARgamma) in the liver of rats with fatty liver disease (FLD) and to explore the role of PPARgamma in the pathogenesis of FLD to provide the basis for using PPARgamma ligand to treat patients with FLD. METHODS: Forty Wistar rats were divided into 4 groups of ten rats each randomly: normal group (group A), alcohol group (group B), fat-rich diet group (group C), alcohol and fat-rich diet group (group D). The rats were sacrificed at the end of the 16th week from the feeding day. Alanine aminotransferase (ALT), tumor necrosis factor-alfa (TNFalpha) in serum and malondialdehyde (MDA) in liver homogenate were determined; livers were collected for observing pathologic changes by HE, Sudan IV, Masson stain under microscope. The morphologic results were analyzed by picture quantitative analysis technique. The changes of ultrastructure were also examined under electron microscope. The expression of PPARgamma in liver was detected by immunohistochemistry and RT-PCR. The correlations between the expression of PPARgamma and biochemical indexes, and liver histology were analyzed. RESULTS: The steatosis, inflammation, necrosis and fibrosis were present in livers of different experimental groups, especially in livers of alcohol and fat-rich diet group. The content of immunodetectable PPARgamma was decreased remarkably in the livers of model rats (group B-D); the level in alcohol and fat-rich diet group (3.43+/-1.48) was significantly lower than that in normal group (18.34+/-3.73), alcohol group (8.82+/-2.52) and fat-rich diet group (11.73+/-2.51) (all P<0.01). The level of PPARgamma mRNA was also lower in the livers of model rats (group B D) than in livers of controls. The expression of PPARgamma in rat liver correlated negatively with the degree of its inflammation, necrosis and fibrosis, as well as the level of serum TNFalpha and the content of MDA in liver homogenates, but not with steatosis or serum ALT. CONCLUSION: Decreased expression of PPARgamma may play an important role in the development of hepatocellular inflammation, necrosis and fibrosis of rats with FLD. Thus, activating PPARgamma by its ligand can be anticipated to provide a therapy target for FLD. PMID- 15112353 TI - Dexamethasone and dextran 40 treatment of 32 patients with severe acute pancreatitis. AB - AIM: Based on the pathogenesis of severe acute pancreatitis and our experimental studies, to investigate the effect of dexamethasone and dextran in treatment of patients with severe acute pancreatitis. METHODS: Thirty-two patients with severe acute pancreatitis were treated with 0.5-1 mg/kg per day dexamethasone for 3-5 d, and 500-1,000 mL/d of dextran 40 for 7 d, besides the routine therapy. RESULTS: After 4-8 h of treatment, abdominal pain began to be relieved; range of tenderness began to be localized in 27 patients. They were cured with nonsurgical treatment. Five of them were deteriorated, and treated with surgery. Four patients in this group died. CONCLUSION: Dexamethasone and dextran 40 block the pathologic process of severe acute pancreatitis through inhibition of inflammatory mediators and improvement of microcirculation disorders respectively. PMID- 15112354 TI - Detection of K-ras point mutation and telomerase activity during endoscopic retrograde cholangiopancreatography in diagnosis of pancreatic cancer. AB - AIM: To study the value of monitoring K-ras point mutation at codon 12 and telomerase activity in exfoliated cells obtained from pancreatic duct brushings during endoscopic retrograde cholangiopancreatography (ERCP) in the diagnosis of pancreatic cancer. METHODS: Exfoliated cells obtained from pancreatic duct brushings during ERCP were examined in 27 patients: 23 with pancreatic cancers, 4 with chronic pancreatitis. K-ras point mutation was detected with the polymerase chain reaction and restriction fragment-length polymorphism (PCR-RFLP). Telomerase activity was detected by PCR and telomeric repeat amplification protocol assay (PCR-TRAP-ELISA). RESULTS: The telomerase activities in 27 patients were measured in 21 exfoliated cell samples obtained from pancreatic duct brushings. D450 value of telomerase activities in pancreatic cancer and chronic pancreatitis were 0.446+/-0.27 and 0.041+/-0.0111, respectively. Seventy seven point eight percent (14/18) of patients with pancreatic cancer and none of the patients with chronic pancreatitis showed telomerase activity in cells collected from pancreatic duct brushings when cutoff value of telomerase activity was set at 2.0. The K-ras gene mutation rate (72.2%) in pancreatic cancer was higher than that in chronic pancreatitis (33.3%) (P<0.05). In considering of both telomerase activities and K-ras point mutation, the total positive rate was 83.3%(15/18), and the specificity was 100%. CONCLUSION: Changes of telomerase activities and K-ras point mutation at codon 12 may be an early event of malignant progression in pancreatic cancer. Detection of telomerase activity and K-ras point mutation at codon 12 may be complementary to each other, and is useful in diagnosis of pancreatic cancer. PMID- 15112355 TI - Epidemiology and outcome of Crohn's disease in a teaching hospital in Riyadh. AB - AIM: To know the epidemiology and outcome of Crohn's disease at King Khalid University Hospital, Riyadh, Saudi Arabia and to compare the results from other world institutions. METHODS: A retrospective analysis of patients seen for 20 years (between 1983 and 2002). Individual case records were reviewed with regard to history, clinical, findings from colonoscopy, biopsies, small bowel enema, computerized tomography scan, treatment and outcome. RESULTS: Seventy-seven patients with Crohn's disease were revisited, 13% presented the disease in the first 10 years and 87% over the last 10 years. Thirty-three patients (42.9%) were males and 44 (57.1%) were females. Age ranged from 11-70 years (mean of 25.3+/ 11.3 years). Ninety-two (92%) were Saudi. The mean duration of symptoms was 26+/ 34.7 mo. The mean annual incidence of the disease over the first 10 years was 0.32:100,000 and 1.66:100,000 over the last 10 years with a total mean annual incidence of 0.94:100,000 over the last 20 years. The chief clinical features included abdominal pain, diarrhea, weight loss, anorexia, rectal bleeding and palpable mass. Colonoscopic findings were abnormal in 58 patients (76%) showing mostly ulcerations and inflammation of the colon. Eighty nine percent of patients showed nonspecific inflammation with chronic inflammatory cells and half of these patients revealed the presence of granulomas and granulations on bowel biopsies. Similarly, 69 (89%) of small bowel enema results revealed ulcerations (49%), narrowing of the bowel lumen (42%), mucosal thickening (35%) and cobblestone appearance (35%). CT scan showed abnormality in 68 (88%) of patients with features of thickened loops (66%) and lymphadenopathy (37%). Seventy-eight percent of patients had small and large bowel disease, 16% had small bowel involvement and only 6% had colitis alone. Of the total 55 (71%) patients treated with steroids at some point in their disease history, a satisfactory response to therapy was seen in 28 patients (51%) while 27 (49%) showed recurrences of the condition with mild to moderate symptoms of abdominal pain and diarrhea most of which were due to poor compliance to medication. Seven patients (33%) remained with active Crohn's disease. Nine (12%) patients underwent surgery with resections of some parts of bowel, 2 (2.5%) had steroid side effects, 6 (8%) with perianal Crohn's disease and five (6.5%) with fistulae. CONCLUSION: The epidemiological characteristics of Crohn's disease among Saudi patients are comparable to those reported from other parts of the world. However the incidence of Crohn's disease in our hospital increased over the last 10 years. The anatomic distribution of the disease is different from other world institutions with less isolated colonic affection. PMID- 15112356 TI - Catheter-related infection in gastrointestinal fistula patients. AB - AIM: To study the incidence, bacterial spectrum and drug sensitivity of catheter related infection (CRI) in gastrointestinal fistula patients. METHODS: A total of 216 patients with gastrointestinal fistulae during January 1998 to April 2001 were studied retrospectively. Two hundred and sixteen catheters of the 358 central venous catheters used in 216 gastrointestinal fistula patients were sent for microbiology analysis. RESULTS: Ninety-five bacteria were cultivated in 88 catheters (24.6%). There were 54 Gram-negative bacteria (56.8%), 35 Gram-positive bacteria (36.8%), and 6 fungi (6.4%). During the treatment of CRI, 20 patients changed to use antibiotics or antifungal, and all patients were cured. The mean time of catheters used was 16.9+/-13.0 d. CONCLUSION: CRI is still the common complication during total parenteral nutrition (TPN) treatment in patients with gastrointestinal fistulae, and Gram-negative bacteria are the main pathogens, and bacterial translocation is considered the common reason for CRI. PMID- 15112357 TI - Capsule endoscopy in diagnosis of small bowel Crohn's disease. AB - AIM: To evaluate the effectiveness of wireless capsule endoscopy in patients with suspected Crohn's disease (CD) of the small bowel undetected by conventional modalities, and to determine the diagnostic yield of M2A Given Capsule. METHODS: From May 2002 to April 2003, we prospectively examined 20 patients with suspected CD by capsule endoscopy. The patients had the following features: abdominal pain, weight loss, positive fecal occult blood test, iron deficiency anaemia, diarrhoea and fever. All the patients had normal results in small bowel series (SBS) and in upper and lower gastrointestinal endoscopy before they were examined. Mean duration of symptoms before diagnosis was 6.5 years. RESULTS: Of the 20 patients, 13 (65%) were diagnosed as CD of the small bowel according to the findings of M2A Given Capsule. The findings detected by the capsule were mucosal erosions (2 patients), aphthas (5 patients), nodularity (1 patient), large ulcers (2 patients), and ulcerated stenosis (3 patients). The distribution of the lesions was mainly in the distal part of the small bowel, and the mild degree of lesions was 54%. CONCLUSION: Wireless capsule endoscopy is effective in diagnosing patients with suspected CD undetected by conventional diagnostic methods. It can be used to detect early lesions in the small bowel of patients with CD. PMID- 15112358 TI - Plasma erythropoietin levels in anaemic and non-anaemic patients with chronic liver diseases. AB - AIM: To investigate the serum erythropoietin (Epo) levels in patients with chronic liver diseases and to compare to subjects with iron-deficiency anaemia and healthy controls. METHODS: We examined 31 anaemic (ALC) and 22 non-anaemic (NALC) cirrhotic patients, 21 non- anaemic subjects with chronic active hepatitis (CAH), 24 patients with iron-deficiency anaemia (ID) and 15 healthy controls. Circulating Epo levels (ELISA; R and D Systems, Europe Ltd, Abingdon, UK) and haemoglobin (Hb) concentration were determined in all subjects. RESULTS: Mean+/ SD of Epo values was 26.9+/-10.8 mU/mL in ALC patients, 12.5+/-8.0 mU/mL in NALC subjects, 11.6+/-6.3 mU/mL in CAH patients, 56.4+/-12.7 mU/mL in the cases of ID and 9.3+/-2.6 mU/mL in controls. No significant difference (P>0.05) was found in Epo levels between controls, CAH and NALC patients. ALC individuals had higher Epo levels (P<0.01) than these groups whereas ID subjects had even higher levels (P<0.001) than patients suffering from ALC. CONCLUSION: Increased Epo values in cirrhotics, are only detectable when haemoglobin was lesser than 12 g/dL. Nevertheless, this rise in value is lower than that observed in anaemic patients with iron-deficiency and appears blunted and inadequate in comparison to the degree of anaemia. PMID- 15112359 TI - Effect of cyclin G2 on proliferative ability of SGC-7901 cell. AB - AIM: To study the effect of cyclin G2 on proliferation of gastric adenocarcinoma cell line-SGC-7901 cell in vitro. METHODS: By use of cation lipofectamine transfection reagent, the pIRES-G2 and pIRESneo plasmids were transferred into SGC-7901cell line. Anticlones were selected by G418. Positive clones were observed and counted using Giemsa staining. Cell proliferative ability was assayed by MTT. RESULTS: (1) The clone number of pIRES-G2 group decreased, clone volume reduced. The number of cell clones in pIRESneo group was 87+/-3, that of pIRES-G2 group was 53+/-4, occupying 60.1% of pIRESneo group, there was significant difference obviously (P<0.01, t=15.45). (2) The average absorbance of clone cell obtained by stable transfection of pIRES-G2 at 570 nm was 1.6966+/ 0.2125, the average absorbance of clone cell obtained by stable transfection of pIRESneo at 570 nm was 2.1182+/-0.3675, there was significant difference between them (P<0.01, t=3.412). CONCLUSION: Cyclin G2 can inhibit SGC-7901cell proliferative ability obviously, it may be a negative regulator in cell cycle regulation. PMID- 15112360 TI - Expression of tumor related gene NAG6 in gastric cancer and restriction fragment length polymorphism analysis. AB - AIM: NAG6 gene is a novel tumor related gene identified recently. This study was designed to examine the expression of this gene in gastric cancer and corresponding normal tissues, and to investigate its role in the occurrence and development of gastric cancer, also to study if the genetic structure of NAG6 was altered in gastric cancer. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR), Northern blot analysis and dot hybridization were used to compare the expression level of NAG6 gene in 42 cases of gastric cancer tissues with their corresponding normal tissues of the same patients respectively. In addition, restriction fragment length polymorphism (RFLP) analysis was adopted to study if the genetic structure of NAG6 was altered in gastric carcinomas. RESULTS: The expression of NAG6 in 57.1% gastric cancer tissues (25/42) was absent by RT-PCR analysis. The down-regulation rate of NAG6 in gastric cancer tissues was significantly higher than that in corresponding normal tissues (P<0.01). However no correlation between the down-regulation of NAG6 and lymph node and/or distance metastasis was found in this study (P>0.05). Dot hybridization confirmed the results of RT-PCR. Furthermore, the results of EcoRI RFLP analysis of NAG6 gene demonstrated that 3 of 7 cases of gastric cancer showed loss of 5 kb fragment in comparison with their corresponding normal tissues. CONCLUSION: NAG6 gene is significantly down regulated in gastric cancer. The loss of genetic materials may be the cause of down-regulation of NAG6 expression. This seems to suggest that NAG6 may represent a candidate of putative tumor suppressor gene at 7q31-32 loci associated with gastric carcinoma. The down regulation of this gene may play a role in occurrence and development of this disease, however it may not be associated with lymph node and/or distance metastasis. PMID- 15112361 TI - Establishment of a P-glycoprotein substrate screening model and its preliminary application. AB - AIM: To establish a high P-glycoprotein (P-gp) expressing cell line as a model for studying drug absorption and distribution, and to explore the preliminary application of this screening model. METHODS: A full-length MDR1 cDNA fragment in plasmid pMDRA1 was first subcloned into plasmid pET28a(+), then MDR1 cDNA was cut from the recombinant plasmid with double-digestion and ligated into the mammalian expression vector pcDNA3.1(+). The recombinant plasmid pcDNA3.1(+)/MDR1 was transfected into breast cancer cell line Bcap37 using the Superfect transfection reagent. Several stably transfected clones were obtained after selection with G418. Real-time fluorescent quantitative RT- PCR and Western blot methods were used to detect the expression of P-gp, and the cellular location of the expressed protein was determined by immunohistochemical staining. Drug sensitivity assay was used to evaluate the biological function of expressed P-gp. Concentration of quercetin in cells was determined by high-performance liquid chromatography (HPLC). RESULTS: The recombinant plasmid was confirmed to be inserted in the correct orientation by restrictive enzyme digestion and DNA sequencing. Real-time fluorescent quantitative RT-PCR showed a higher level of P-gp mRNA in transfected cells compared to that in the control cells, and the Western blot result also indicated that P-gp expression in transfected cells was higher than that in control cells. The immunohistochemical staining showed that the expressed P-gp was localized on cell membranes. Drug sensitivity assay showed that the IC50 for adriamycin and colchicine of the transfected cells was higher than that of the control cells. The concentration of quercetin in model cells was lower than that in control cells by HPLC. After P-gp inhibitor verapamil was administered, the concentration of quercetin in model cells was increased. CONCLUSION: A high P-gp expressing cell line can be established, which could provide a suitable in vitro model system for studying drug intestinal absorption mechanism, predicting the drug permeability characteristics and screening new multi-drug resistance reversing agents. With this model, quercetin can be found to be transported by P gp, and it is a P-gp substrate. PMID- 15112362 TI - Diagnosis of intestinal acariasis with avidin-biotin system enzyme-linked immunosorbent assay. AB - AIM: To explore the value of avidin-biotin system enzyme-linked immunosorbent assay (ABC-ELISA) in diagnosis of intestinal acariasis. METHODS: Mite-specific IgG levels in serum of 48 patients with intestinal acariasis were measured with ABC-ELISA. The sensitivity of this method was compared with that of staphylococcal protein A enzyme-linked immunosorbent assay (SPA-ELISA). RESULTS: The positive rate of mite-specific IgG detected with ABC-ELISA and SPA-ELISA was 89.58% (43/48) and 56.25% (27/48), respectively. The positive rate with ABC-ELISA was statistically higher than that with SPA-ELISA (chi2=13.50, P<0.01). CONCLUSION: ABC-ELISA is an effective method for the diagnosis of intestinal acariasis. PMID- 15112363 TI - Catheterization-associated complications of intraperitoneal chemotherapy in advanced gastric cancer. AB - AIM: To assess the catheterization-associated complications during intraperitoneal chemotherapy (IPCT) for advanced gastric cancer. METHODS: From 1998 to 2002, 80 patients with advanced gastric cancer received a total of 320 courses of IPCT using a large bore central venous catheter and associated complications were analyzed. RESULTS: Catheterization-associated complications occurred in 11 of the 80 patients (13.8%), including abdominal pain caused by catheter in 2 cases (0.63%), insertion failure in 2 cases (0.63%), bowel perforation in 1 case (0.31%) and abdominal pain during chemotherapy in 6 cases (1.88%). No serious complications required surgical intervention. CONCLUSION: IPCT using central venous catheters can be performed safely and simply without severe associated complications. PMID- 15112365 TI - Evaluation of effect of hybrid bioartificial liver using end-stage liver disease model. AB - AIM: To study the role of hybrid bioartificial liver (HBL) in clearing proinflammatory cytokines and endotoxin in patients with acute and sub-acute liver failure and the effects of HBL on systemic inflammatory syndrome (SIRS) and multiple organ dysfunction syndrome (MODS). METHODS: Five cases with severe liver failure (3 acute and 2 subacute) were treated with HBL. The clinical signs and symptoms, total bilirubin (TBIL), serum ammonia, endotoxin TNF-alpha, IL-6 and prothrombin activity (PTA),cholinesterase (CHE) were recorded before, during and after treatment. The end-stage liver disease (MELD) was used for the study. RESULTS: Two patients were bridged for spontaneous recovery and 1 patient was bridged for OLT successfully. Another 2 patients died on d 8 and d 21. The spontaneous recovery rate was 30.0%. PTA and CHE in all patients were significantly increased (P<0.01), while the serum TBIL, endotoxin, TNF-alpha, IL 6 were decreased. MELD score (mean 43.6) predicted 100% deaths within 3 mo before treatment with HBL. After treatment with HBL, four out of 5 patients had decreased MELD scores (mean 36.6). The MELD score predicted 66% mortalities. CONCLUSION: The proinflammatory cytokines (TNFalpha, IL-6 and endotoxin)can be significantly removed by hybrid bioartificial liver and HBL appears to be effective in blocking SIRS and MODS in patients with acute and sub-acute liver failure. MELD is a reliable measure for predicting short-term mortality risk in patients with end-stage liver disease. The prognostic result also corresponds to clinical outcome. PMID- 15112364 TI - Protective effect of doxorubicin induced heat shock protein 72 on cold preservation injury of rat livers. AB - AIM: To observe the protective effect of heat shock protein 72 (HSP 72) induced by pretreatment of doxorubicin (DXR) on long-term cold preservation injury of rat livers. METHODS: Sprague-Dawley rats were administered intravenously DXR at a dose of 1 mg/kg body mass in DXR group and saline in control group. After 48 h, the rat liver was perfused with cold Linger's and University of Wisconsin (UW) solutions and then was preserved in UW solution at 4 degrees for 24, 36 and 48 h. AST, ALT, LDH and hyaluronic acid in preservative solution were determined. Routine HE, immunohistochemical staining for HSP 72 and electron microscopic examination of hepatic tissues were performed. RESULTS: After 24, 36 and 48 h, the levels of AST, ALT and hyaluronic acid in preservative solution were significantly higher in control group than in DXR group (P<0.05), while LDH level was not significantly different between the 2 groups (P>0.05). Hepatic tissues in DXR group were morphologically normal and significantly injured in control group. HSP 72 was expressed in hepatocytes and sinusoidal endothelial cells in DXR group but not in control group. CONCLUSION: Pretreatment of DXR may extend the time of rat liver cold preservation and keep liver alive. The expression of HSP 72 in liver can prevent hepatocytes and sinusoidal endothelial cells from long-term cold preservation injury. PMID- 15112366 TI - Expression and significance of angiopoietin-2 in gastric cancer. AB - AIM: To investigate the expression and pathological factors of Angiopoietin-2 (Ang-2) in primary gastric cancers and adjacent normal tissues. METHODS: The expression of Angiopoietin-2 and VEGF were studied in 72 primary gastric cancers and adjacent normal tissues from the same patients by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry. RESULTS: Ang-2 was mainly expression in tumor cells. There were significantly difference between expression of Ang-2 in primary gastric cancer and in adjacent normal tissue samples (P=0.003). It was statistically correlation between Ang-2 and VEGF expression in tumors (P=0.0055). With regard to Ang-2 expression in tumors, there were significant difference between early stage and advanced stage (P=0.017), and significant difference between positive vascular involvement and negative vascular involvement (P=0.032). However, there was no significant difference between moderate-poor differential type and high differential type (P=0.908), between positive lymph node metastasis and negative lymph node metastasis (P=0.752), between positive serosal invasion and negative serosal invasion (P=0.764). The cases with expression of Ang-2 were increasing with advanced stage and vascular involvement. CONCLUSION: The results manifested that Angiopoietin-2, coordinated with VEGF, play role in regulating tumor angiogenesis of gastric cancer. PMID- 15112367 TI - Gaussian processes for machine learning. AB - Gaussian processes (GPs) are natural generalisations of multivariate Gaussian random variables to infinite (countably or continuous) index sets. GPs have been applied in a large number of fields to a diverse range of ends, and very many deep theoretical analyses of various properties are available. This paper gives an introduction to Gaussian processes on a fairly elementary level with special emphasis on characteristics relevant in machine learning. It draws explicit connections to branches such as spline smoothing models and support vector machines in which similar ideas have been investigated. Gaussian process models are routinely used to solve hard machine learning problems. They are attractive because of their flexible non-parametric nature and computational simplicity. Treated within a Bayesian framework, very powerful statistical methods can be implemented which offer valid estimates of uncertainties in our predictions and generic model selection procedures cast as nonlinear optimization problems. Their main drawback of heavy computational scaling has recently been alleviated by the introduction of generic sparse approximations.13,78,31 The mathematical literature on GPs is large and often uses deep concepts which are not required to fully understand most machine learning applications. In this tutorial paper, we aim to present characteristics of GPs relevant to machine learning and to show up precise connections to other "kernel machines" popular in the community. Our focus is on a simple presentation, but references to more detailed sources are provided. PMID- 15112368 TI - An annealed chaotic maximum neural network for bipartite subgraph problem. AB - In this paper, based on maximum neural network, we propose a new parallel algorithm that can help the maximum neural network escape from local minima by including a transient chaotic neurodynamics for bipartite subgraph problem. The goal of the bipartite subgraph problem, which is an NP- complete problem, is to remove the minimum number of edges in a given graph such that the remaining graph is a bipartite graph. Lee et al. presented a parallel algorithm using the maximum neural model (winner-take-all neuron model) for this NP- complete problem. The maximum neural model always guarantees a valid solution and greatly reduces the search space without a burden on the parameter-tuning. However, the model has a tendency to converge to a local minimum easily because it is based on the steepest descent method. By adding a negative self-feedback to the maximum neural network, we proposed a new parallel algorithm that introduces richer and more flexible chaotic dynamics and can prevent the network from getting stuck at local minima. After the chaotic dynamics vanishes, the proposed algorithm is then fundamentally reined by the gradient descent dynamics and usually converges to a stable equilibrium point. The proposed algorithm has the advantages of both the maximum neural network and the chaotic neurodynamics. A large number of instances have been simulated to verify the proposed algorithm. The simulation results show that our algorithm finds the optimum or near-optimum solution for the bipartite subgraph problem superior to that of the best existing parallel algorithms. PMID- 15112369 TI - Predicting spatial data with RBF networks. AB - Spatial prediction needs to account for spatial information, which makes conventional radial basis function (RBF) networks inappropriate, for they assume independent and identical distribution. In this paper, we fuse spatial information at different layers of RBF. Experiments show fusion at hidden layer gives the best result and suggest that the optimal value is around one for the coefficient, which is used in the linear combination at the output layer. PMID- 15112370 TI - Nonlinear system modelling via optimal design of neural trees. AB - This paper introduces a flexible neural tree model. The model is computed as a flexible multi-layer feed-forward neural network. A hybrid learning/evolutionary approach to automatically optimize the neural tree model is also proposed. The approach includes a modified probabilistic incremental program evolution algorithm (MPIPE) to evolve and determine a optimal structure of the neural tree and a parameter learning algorithm to optimize the free parameters embedded in the neural tree. The performance and effectiveness of the proposed method are evaluated using function approximation, time series prediction and system identification problems and compared with the related methods. PMID- 15112371 TI - Fractal characterization of bpn weights evolution. AB - Training methodology of the Back Propagation Network (BPN) is well documented. One aspect of BPN that requires investigation is whether or not the BPN would get trained for a given training data set and architecture. In this paper the behavior of the BPN is analyzed during its training phase considering convergent and divergent training data sets. Evolution of the weights during the training phase was monitored for the purpose of analysis. The evolution of weights was plotted as return map and was characterized by means of fractal dimension. This fractal dimensional analysis of the weight evolution trajectories is used to provide a new insight to understand the behavior of BPN and dynamics in the evolution of weights. PMID- 15112372 TI - Is metastatic potential of breast cancer inherent or acquired? PMID- 15112373 TI - Ethical decision making within the bureaucratic context: a case study. AB - Balancing the polarized notions of quality and quantity of service and care is a challenge for the 21st century health and social service system. The expectation of transparent decision making at both the policy and the practice level of case management has forced practitioners to seek guidance in ethical decision making. Case management, as a system, has potential to lead this new practice approach by incorporating the principles of ethical decision making in planning and coordinating care. A recent case study of ethical decision making within the bureaucratic context offers some insight and learning and may help inform future practice. PMID- 15112374 TI - Community treatment and rehabilitation: one agency's model of case management. AB - This article describes one agency's model of delivering case management services to an adult population of seriously and persistently mentally ill persons. It was developed following improvements in psychotropic medications that have resulted in considerable symptom relief for many clients. The treatment program, both medical and psychosocial aspects, is focused on recovery rather than maintenance. Outcome measures of treatment and rehabilitation objectives demonstrated that the program has been successful. PMID- 15112375 TI - Instituting a case management system for the lead-poisoned child: the Maine experience. AB - This article details the origin and implementation of a state-level case management system for children with elevated blood lead levels. This includes a thorough accounting of the methods utilized to create the system. The article closes with data derived from the case management reporting system showing the system's effectiveness and with a description of how this design parallels the case management recommendations by the Centers for Disease Control and Prevention made at a later date. The article offers suggestions for designers of such systems. In general, the authors advise building on existing systems, creating written protocols for both case managers and health care personnel, developing a support system for case managers, tracking activities through data, and giving recognition to field personnel. PMID- 15112376 TI - Quality assurance practices in care management: a perspective from the United Kingdom. AB - Alongside the development of care management in many countries has emerged a need to monitor the quality of both care management practices, as well as the quality of services provided as a consequence of care planning by care managers. Designing and implementing quality assurance practices in care management present special challenges. This article examines current practices from the perspective of the United Kingdom (UK). First, current performance initiatives at the central government level are described, including an indication of their limitations in judging quality in care management; second, existing work on assuring quality in care management is reviewed along with its relevance for the UK; and third, a local study, identifying measures within an analytical framework and using them to monitor care management against set dimensions, is described. Parallel developments are now occurring at both national and local levels in the UK, with local work both influencing and being informed by national developments. Future issues to address include the problems in collecting relevant data and in extending methods of analysis to monitor care management more comprehensively. PMID- 15112377 TI - A survey of family care giving to elders in New York State: findings and implications. AB - It is estimated that there are 734,400 care giver households in New York State (9.6% [+/-] 0.8% of all households). Categorization of all care givers on a 5 level "intensity of care" measure reveals that, on average, care givers provide 22.1 hours of care per week. Highest intensity level 5 care givers (9.2% of all care givers), provide, on average, 88 hours of care per week and account for 36.3% of all care giving. The annualized market contribution of all care givers to the NYS health care system is estimated at between $7.5 and $11.2 billion dollars. The combination of care levels 4 and 5 contributed 70% of all care giving and account for about $5.2 billion in market value. Level 4 and 5 care givers are more likely than other care givers to report difficulties including the recent death of the care-receiver (p < 0.001), financial and employment setbacks (p < 0.001), emotional stress from worrying about the patient's future condition, dealing with cognitively impaired, physically unmanageable, opinionated, virtually immobile patient's, and insufficient support from family members (p < 0.05). Nevertheless, they also report such rewards as feeling grateful for improving the quality of the care receiver's life (43.5%) as well as love (17.2%). Of the nearly 15% of NYS care givers who used adult day care services, all reported that these services met their needs fully or partly. However, of the 85% who did not use the service, 33.5% were negative about it. And while use of adult day care services increases with intensity of care level, there is resistance to adult day care particularly among level 3 care givers, with negative statements from 46.8% of them. PMID- 15112378 TI - Medicare managed care and frail elders: lessons from social HMOs. AB - After 20 years of operations, the Social HMO demonstration of integrated acute and long-term care is scheduled to end on December 31, 2004. While a new disability adjustment to the Medicare payment system promises to provide the financial underpinning for continuing to serve the 113,000 beneficiaries now enrolled at four sites, a broader regulatory structure as an alternative to current waivers is also needed. The regulations could also accommodate other frail elderly programs, which serve nursing home residents and beneficiaries of both Medicare and Medicaid. The relevance of Social HMOs--the largest and most broadly targeted of frail elderly programs--is reviewed herein, particularly regarding marketing, selectivity, reimbursement, and special frail elderly benefits and geriatric services. PMID- 15112379 TI - Immunohistochemical detection of acetylation and phosphorylation of histone H3 in cervical smears. AB - OBJECTIVE: Histones bind in a sequence-independent manner to form chromatin. The aminoterminal tails of histones are targets for both phosphorylation and acetylation events. These modifications are thought to fundamentally regulate chromatin structure to accommodate transcription, DNA replication, mitosis and DNA repair. Regeneration of squamous epithelium is accompanied by marked cellular atypia, nuclear and nucleolar pleomorphism which could be confused with neoplasia. The aim of the study was to detect phosphorylated and acetylated forms of histone H3 in cytological smears. DESIGN: Translational research. SETTING: Department of Experimental Oncology, Masaryk Memorial Cancer Institute, Department of Pathological Physiology, Medical Faculty and Department of Obstetrics and Gynecology, Masaryk University, Czech Republic. METHODS: Smears from women aged between 20 to 46 years were selected. The specimens comprised 10 squamous metaplasia, 20 CIN I, 12 CIN II, and 14 CIN III. The smears were stained with polyclonal antibodies against phosphorylated and acetylated forms of histone H3. RESULTS: We found that nuclear positivity for phosphorylated (P) and acetylated (A) forms of histone H3 in CIN II (23% P, 33% A) and CIN III (25% P, 44% A) was higher in comparison with CIN I (8% P, 15% A) and metaplasia (11% P, 12% A). CONCLUSION: We revealed a marked association of histone H3 modifications with the progression of CIN II, CIN III in comparison with CIN I and metaplasia. Our results are in agreement with recent findings: 1. staining of cells with anti phospho-histone H3 antibodies therefore provides a highly specific marker for mitosis. 2. acetylation of nucleosomal histones correlates with localised transcriptional activity. PMID- 15112380 TI - [Microbiological findings in patients with recurrent vulvovaginal candidiasis in the Hradec Kralove Faculty Hospital 1995-2002]. AB - OBJECTIVE: To evaluate the microbiological findings in the patients with the recurrent vulvovaginal candidiasis (RVVC) with a focus on the establishment of fungal etiology and its in vitro antifungal susceptibility. DESIGN: Retrospective clinical and laboratory study. SETTING: Department of Obstetrics and Gynecology, Medical Faculty Hradec Kralove, Charles University, Prague, Department of Clinical Microbiology, Medical Faculty Hradec Kralove, Charles University, Prague, Department of Biological and Medical Sciences, Faculty of Pharmacy Hradec Kralove, Charles University, Prague. METHODS: An analysis of clinical and anamnestic data in outpatients of the Dept. of Obstetrics and Gynecology and the laboratory data from the microbiological examinations performed in the Dept. of Clinical Microbiology from 1995 to 2002. RESULTS: Candida albicans accounted for 88.5% of the episodes of RVVC in the setting of 56 patients. Non-albicans Candida species were represented especially by C. glabrata (4.9%) and C. krusei (3.1%). There were no considerable differences between the spectrum of RVVC and acute vulvovaginal candidiasis with the exception of Saccharomyces cerevisiae (0.7% in RVVC vs. 3.7% in acute VVC). Mycological findings in 61 (20.5%) samples were accompanied by bacterial microbiota with the predominance of Streptococcus agalactiae (n = 15) and Gardnerella vaginalis (n = 9). Decreased antifungal susceptibility determined by the disk test was observed in the strains of C. glabrata, C. krusei and S. cerevisiae, the other yeast isolates being susceptible to all ten antifungal drugs tested. CONCLUSION: The microbiological examination was decisive for the establishment of the diagnosis of RVVC in most cases. The most frequent etiological agents responsible for the attacks of RVVC as well as for acute vulvovaginal candidiasis was C. albicans, which was generally susceptible to antifungal drugs. PMID- 15112381 TI - [Comparison of apoptosis expression in the secretory endometrium during a normal menstrual cycle]. AB - OBJECTIVE: Comparison of level of apoptosis in the mid-secretory human endometrium. DESIGN: Clinical-laboratory trial. SETTING: Department of Gynecology and Obstetrics, Medical Faculty, Palacky University, Faculty Hospital, Olomouc, Department of Gynecology and Obstetrics, Bata Hospital, Zlin. METHODS: Samples of secretory endometrium were obtained from 14 women. The women included in the study had the next criteria: a history of infertility of more than 12 months, regular menstrual cycles, age below 40 and FSH basal level range 5-9 IU/L. Sampling of tissue was performed after confirmed ovulation by LH surge and repeated ultrasound examinations. Patterns were processed by routine methods and apoptosis was detected using TUNEL assay. RESULTS: The level of apoptosis in the endometrial epithelial surface was significantly higher on day +7 compared to day after ovulation (P < 0.02). Apoptotic cells were seen only sporadically in endometrial stromal cells. The level of apoptosis in endometrial stroma was also higher (but not statistically significant) on day +7 compared to day The level of apoptosis was markedly higher in spongy layer on day +7 (P < 0.01) compared to day There were no significant differences between day and +7 in compact layers of the glands. PMID- 15112382 TI - [Effect of toxic substances in the environment on reproduction]. AB - OBJECTIVE: To summarize the findings about the influence of some xenobiotics on human reproduction, especially infertility. TYPE OF STUDY: Review article. SETTING: The Department of Obstetrics and Gynecology, 1st Medical Faculty, Charles University and the General Faculty Hospital, Prague METHODS: Analysis of literature. RESULTS: A review about the influence of some xenobiotics on human reproduction was given. In some there is a possible negative influence on the results of in vitro fertilisation program. It is a question of hormone disruptors, some heavy metals, some organic solvents, pesticides and smoking of cigarettes. CONCLUSION: In most of these xenobiotics there is direct evidence of their negative influence on reproduction and the results of in vitro fertilisation program. Therefore it is worth to continue in this evaluation. PMID- 15112383 TI - [Classification of twins and their ultrasonographic diagnosis]. AB - OBJECTIVE: Systematic classification of all forms of twins and mechanisms of twinning. DESIGN: Review of published cases. Morphogenesis based on personal experience related to human blastogenesis including observation of anterior twinning in two early human embryos. SETTING: Institute for the Care of Mother and Child, Prague, Department of Gynecology and Obstetrics, 1st Medical Faculty, Charles University, Prague, Department of Medical Genetics, Medical Faculty, Palacky University Olomouc, Institut Unica, Brno. METHODS: Analysis of cases described in literature completed by observed cases. RESULTS: Classification of twins. A) Separated twins: 1. dichorial a) monozygotic (very rare), b) dizygotic (most frequent); 2. monochorial (always monozygotic) a) diamnial, b) monoamnial. B) Conjoined twins (always monozygotic) 1. isopagi (equal conjoined twins) a) originating from peripheral fusions of two germ discs, b) originating from duplications of axial structures; 2. heteropagi (unequal conjoined twins): autosit (main twin), heterosit (parasitic twin). CONCLUSION: The developmental mechanisms of twinning are discussed, special attention is paid to equal conjoined twins and to the possibilities of their early prenatal ultrasonographic diagnostics. PMID- 15112384 TI - [Is the information we give women prior to invasive prenatal procedures adequate?]. AB - BACKGROUND: The aim was to evaluate the psychological impact on women undergoing invasive procedures of prenatal diagnosis. SETTING: Department of Gynecology and Obstetrics, 1st Medical Faculty, Charles University, Prague. MATERIAL AND METHODS: A questionnaire was given to 200 pregnant women and to 160 midwives and students. The acquired data were statistically evaluated using the non-parametric chi 2 test for a 5% confidence interval and the Kruskal-Wallis test (analysis of non-normal distribution of random variables). RESULTS: We found that 85% of pregnant patients were satisfied with the information given by their obstetrician prior to the procedure, 53% of the patients were distressed about the procedure. The largest percentage of patients feared complications of the procedure, while fear of the results of the procedure took second place. CONCLUSION: We found that only some of the patients and midwives had complete information about the actual method of performing these procedures, about the risk, and about the time it takes to obtain results. Most patients receive their information from a doctor geneticist, which is in agreement with our system. The patient's distress regarding the procedure is not dependent on the level of education. From the acquired data, it follows that greater significance should be placed on the informing patients as well as midwives about all aspects of performing invasive procedure of prenatal diagnosis. According to our study, neither the patient nor the midwife have an adequate perception of the benefits and risk of prenatal diagnostic examinations. PMID- 15112385 TI - [Effect of early cleavage of embryos after intracytoplasmic sperm injection (ICSI) into oocytes on results of fertilization in vitro (IVF) and embryo transfer (ET)]. AB - OBJECTIVE: To evaluate the influence of early cleavage in embryos obtained using ICSI on the success rate of IVF-ET. DESIGN: A prospective study. SETTING: Centre of Assisted Reproduction, Department of Obstetrics and Gynecology, Medical Faculty, Palacky University, Olomouc. METHODS: The rate of the first cleavage of embryos was checked between 23 to 26 hours after ICSI in 115 cycles IVF-ET. "Early Cleavage" (EC) embryos which were already cleaved were cultivated separately from the "slow" (No Early Cleavage, NEC) embryos. After 3 days cultivation, no more than three 6- to 8-cell embryos were transferred. Only EC embryos were transferred in 43 cycles and only NEC embryos were transferred also in 43 cycles. The remaining 29 cycles, where both EC and NEC embryos had to be transferred, were not evaluated. In both groups the following entry parameters were compared: age of the patient, number of fertilized oocytes, fertilization rate (FR), number of frozen and evaluated embryos, number of EC embryos recorded and number of transferred embryos. The outcome parameters compared were: pregnancy rate (PR), number of gestational sacs with heart beats per number of transferred embryos (implantation rate, IR), abortions (AB) and the frequency of multiple pregnancy. Statistical evaluation was performed by t-test, chi 2 test and by Sheffe analysis. RESULTS: EC embryos were found in 57% of cycles (n = 66) after ICSI. Early cleavage was observed in 198 of 563 embryos (35%). Among entry parameters the only statistically significant differences (P < 0.05) were found in the number of retrieved oocytes as well as in the ratio of frozen embryos which were higher in the cycles with the transfer of only EC embryos. FR was the same (74%) in both types of transfer. Outcome parameters in the transfer of only EC versus NEC embryos were higher in PR (40% vs. 33%), about equal in IR (27% vs. 25%) and lower in AB (2% vs. 5%). Moreover, multiple pregnancies were less frequent in the transfer of only EC embryos: 65% of singleton pregnancy, 35% of twins and no triplet versus 36%, 57% and 7%, resp., in the transfer of only NEC embryos. The differences in outcome parameters are not statistically significant (P > 0.05) because of small numbers. CONCLUSION: The speed of the first cell cleavage is a useful additional criterion for better evaluation of quality and viability of embryos. PMID- 15112386 TI - [Personality and characteristics of couples in infertile marriage]. AB - OBJECTIVE: To estimate the basic personality and couple characteristics of men and women from infertile marriages. DESIGN: Research psychodiagnostical study. SETTING: Institute of Sexology, 1st Medical Faculty, Charles University, Prague, Iscare IVF, Prague. METHODS: 38 couples applying for assisted fertilization were psychologically examined by standard psychodiagnostical methods. RESULTS: Men and women from infertile couples displayed moderate anomal behavioral characteristics, men from these couples are more neurotic than general population, women display more anxiety and social desirability. The relations in these marriages are not substantially disturbed and are even less conflictuous and the consistency of the male and female roles is even better than in general population. Men from infertile couples are perceived by themselves as well as by their wives as less masculine. CONCLUSION: In men and women from examined couples there were no serious personality anomalies and the relations in these marriages were not substantially disturbed by infertility. PMID- 15112387 TI - [Pregnancy after uterine artery embolization in uterine myoma]. AB - OBJECTIVE: To report a case of successful pregnancy outcome after embolization treatment of symptomatic leiomyoma. DESIGN: Case report. SETTING: Department of Gynaecology and Obstetrics of University Hospital, Hradec Kralove. SUBJECT AND METHOD: A 41-year old primigravida who had previously undergone uterine artery embolization and myomectomy had successful course of the spontaneously conceived pregnancy terminated per sectionem cesarean. An analysis of the 50 published cases of pregnancy after uterine artery embolization revealed the following complications: spontaneous abortion (22%), malpresentation (17%), hypotrophy (7%), premature delivery (28%), caesarean section (58%) and postpartum hemorrhage (13%). CONCLUSION: Whether this procedure is safe for women desiring future fertility is controversial. There are very few data regarding the outcomes of pregnancies after embolization. PMID- 15112388 TI - [The first pregnancy and birth after cryopreservation of oocytes in the Czech Republic--importance of the method for reproductive medicine]. AB - OBJECTIVE: The purpose of this work was to develop methods for successful cryopreservation of human oocytes. DESIGN: Case-controlled study and case report. SETTING: Sanatorium Pronatal, Prague. METHODS: Propanediol (PrOH)-sucrose was used as cryoprotectant medium for cumulus-free oocytes and stepwise dilution of cryoprotectant post-thaw. RESULTS: The method was used in three patients (38 denuded oocytes) and yielded excellent survival and fertilization rates (89.5% and 73.5% respectively). In all patients embryo-transfer was possible, one patient got pregnant and delivered a healthy baby. CONCLUSION: Our data show that cryopreservation may ensure that the integrity of the human oocyte is adequate for normal fertilization and embryo development. PMID- 15112389 TI - [Current results in treatment of omphalocele and gastroschisis]. AB - OBJECTIVE: The aim of the study was to evaluate current possibilities of treatment of newborns with abdominal wall defects: omphalocele and gastroschisis. SETTING: Department of Pediatric Surgery, 2nd Medical Faculty Charles University and Faculty Hospital Motol, Prague. METHODS: A retrospective cohort study of the treatment results in newborns with defect of abdominal wall. RESULTS: Thirty eight newborns with omphalocele and gastroschisis were treated at the authors department in the period of 1995-2002. Primary closure of defect was possible in 23 newborns (60%), Goretex patch or "silo" technique was used in 13 patients (34%), while two newborns (6%) were not operated on. The overall survival in children with omphalocele was 73%, being 89% in children without chromosomal anomalies. Six infants died: three with trisomy 13, one with trisomy 18, one infant died of sepsis and one had primary pulmonary hypertension and congenital heart defect. The survival in children with gastroschisis was 94%, one child died of multiorgan failure during sepsis. CONCLUSION: Nowadays prenatal diagnosis of gastroschisis or omphalocele is a recommendation for complex examination and consultation in a specialized center. The prognosis for most isolated defects of abdominal wall is good as far as survival and quality of life are concerned. PMID- 15112390 TI - [Incidence and survival in children with selected types of congenital defects in the Czech Republic from 1994 to 2001. (Part 1)]. AB - OBJECTIVE: Analysis of prenatal and postnatal incidence of selected types of birth defects in the Czech Republic in the period of 1994-2001. In the postnatally diagnosed cases the survival in the course of the first year of life was analyzed. Selected birth defects--anencephaly, spina bifida, encephalocele, neural tube defects, congenital hydrocephalus, Down syndrome, were investigated. TYPE OF STUDY: Retrospective analysis from registry of birth defects in the UZIS Czech Republic in the period of 1994-2001. METHOD: The authors analyzed frequency of prenatally and postnatally diagnosed cases of 6 selected types of birth defects in the period of 1994-2001 in the Czech Republic. In cases where the diagnosis was established after birth, analysis of survival and death rate during the first year of life was performed for selected birth defects. RESULTS: In the period of 1994-2001, 745,410 children were born in the Czech Republic. There were 3,572 children with one or more birth defects diagnosed in this cohort during the first year of life. The mean incidence of all cases in this period of observation was 304.52 per 10,000 live born children. The percentage of prenatally diagnosed cases, ended for this diagnosis in the above mentioned period of observation, was on the average 72.5% in neural tube and the highest in anencephaly--more than 96%. In cases of congenital hydrocephalus it was higher than 45% and in Down syndrome it represented 56.5%. The survival in children with neural tube defects was 82.5% on the average, being obviously zero in anencephaly. The children with congenital hydrocephalus survive in more than 75% and those with Down syndrome survive in almost 95%. CONCLUSION: Incidence of these types of birth defects in newborns in the Czech Republic is presently lower than in previous years particularly due to successful prenatal diagnostics. Cases, where the diagnosis is established after delivery, represent an important part of perinatal, neonatal and post-neonatal mortality and morbidity. PMID- 15112391 TI - [Importance of genetic testing in couples with reproductive disorders]. AB - OBJECTIVE: To determine the prevalence of chromosomal aberrations in infertile couples undergoing in vitro fertilization (IVF). DESIGN: Cytogenetic analysis of peripheral blood lymphocytes in the group of patients undergoing IVF. Detection of chromosomal aberrations in the fetuses after IVF. SETTING: Department of Medical Genetics and Fetal Medicine, Medical Faculty, Palacky University and the University Hospital, Olomouc. METHODS: Cultivation of peripheral blood lymphocytes or fibroblasts of amniotic fluid. Using fluorescent in situ hybridization in cases of mosaicism. RESULTS: Out of 638 patients undergoing treatment for male or female infertility, 595 had normal karyotype and 43 (6.8%) had abnormal karyotype. There were detected 9 (1.4%) cases of balanced chromosomal rearrangements, 2 (0.31%) cases of deletion of Y chromosome, 2 (0.31%) cases of inversion, 2 (0.31%) cases of marker chromosome, 5 (0.78%) cases of gonosomal aneuploidy (47,XXY) and 23 (3.65%) cases of gonosomal mosaicism--out of the 22 (3.5%) cases of low-level mosaicism. In the small group of pregnant patients after IVF investigated for the risk of genetic disorders included in our study (n = 60) the frequency of chromosomal abnormalities was 9 (15%). CONCLUSIONS: Our data show that a high number of infertile couples is affected by chromosomal aberrations which occur more frequently in females than in males. It is caused by high frequency of low-level gonosomal mosaicism in the group of infertile women. Chromosomal analyses are highly recommended before each IVF procedure. PMID- 15112392 TI - [Research design strategy in medical and biostatistical studies]. AB - OBJECTIVE: The purpose of this review is to discuss basic methods of the biostatistical study design and analysis. DESIGN: A review article. SETTING: OBGYN clinic, 1st Medical Faculty, Charles University, Teaching Hospital Bulovka, Prague. SUBJECT: The medical research requires the complexity of special skills, including medical practice, biological understanding and biostatistical expertise. The following article discusses methods of the biostatistical study design and analysis using synoptical clinical and health examples. PMID- 15112393 TI - The importance of early intervention. PMID- 15112394 TI - Patient comfort with resin technology. PMID- 15112395 TI - A simple technique for core buildup. PMID- 15112396 TI - Treatment options for the edentulous mandible: clinical application of the two implant overdenture. AB - Edentulism is a public health concern that affects millions of individuals. While this condition has been treated for years with implant-supported hybrid prostheses, this modality does not always represent the optimal restorative solution for all patients. The two-implant mandibular overdenture provides greater retention than does a conventional mandibular denture and, due to its cost and efficacy, can significantly improve an edentulous patient's quality of life. This article reviews the nutritional and social aspects of edentulism in affected patients and its treatment with two-implant overdentures. PMID- 15112397 TI - A simplified approach in the fabrication of an implant-supported, full-mouth, fixed metal-ceramic restoration. PMID- 15112398 TI - The role of tissue management and gingival recontouring on clinical success. PMID- 15112399 TI - Simplified posterior aesthetics using microhybrid composite resins: techniques for success. AB - Throughout the evolution of composite resins there has been a search for a system that blends aesthetics and strength. Although current resin systems allow the clinician to deliver beautiful results, the multiple layering technique using a variety of different composite resins to create these results may not be practical for the average dentist. This article explores the impact that a microhybrid composite resin can have on predictable and consistent results using a simplified approach. PMID- 15112400 TI - Endodontic root reinforcement using a bonded, fiber-reinforced gutta-percha- tipped obturator. PMID- 15112401 TI - Interdisciplinary treatment of a severe maxillary aesthetic defect: a case report. AB - The treatment of compromised dentition as a result of severe bone deficiencies in the aesthetic zone requires the use of predictable alveolar bone regeneration and restorative techniques. A multidisciplinary treatment approach based on thorough communication and treatment planning will allow the delivery of desirable periodontal and prosthetic results. By considering the patient's desires and existing condition, different treatment options can be presented to determine which best suits the patient's needs. This presentation describes a treatment protocol for restoration of the anterior region following bone augmentation, implant placement, and prosthetic rehabilitation. PMID- 15112402 TI - [The modelling of medical and evacuation support of troops in zone of armed conflict (First report: methodical approach to the development of complex modelling scenario of the system for rendering of specialized medical care)]. PMID- 15112403 TI - [The experience of training conducted with the special medical detachment under conditions of strict antiepidemic regimen]. PMID- 15112404 TI - [Cold trauma in servicemen: comparative characteristic and staged treatment]. PMID- 15112405 TI - [Epidemiologic analysis of non-hospital pneumonia incidence in the troops]. AB - The analysis of dynamics of pneumonia incidence in servicemen for 50 years and 1 year was conducted. The risk factors of pneumonia development in the troops were studied. Some of them decrease the body immunoresistance (supercooling, acute respiratory disease, adaptation to the service and new climatic-and-geographic conditions, stress, body mass deficiency, etc.). The others activate the mechanism of pneumonia agent transmission ("mixing" factor during the fresh forces admission, overcrowding, many people in the sleeping rooms, etc.). To prevent the pneumonia incidence in military collectives the prophylactic and anti epidemic measures should reduce the influence of these factors. PMID- 15112406 TI - [Method of ureteral neoanastomosis formation]. PMID- 15112407 TI - [Anesthesia during cardiosurgical operations based on high epidural blockade combined with pofol]. PMID- 15112408 TI - [Peculiarities of social and psychological adaptation of the wounded servicemen]. PMID- 15112409 TI - [Evaluation of sinus rhythm stability after external electric cardioversion in patients with paroxysmal fibrillar arrhythmia]. PMID- 15112410 TI - [Peculiarities of diagnostics and therapy of atypical pulmonary diseases in Russian hospital in Kosovo]. PMID- 15112411 TI - [Organisation of therapy and vacation for servicemen and their families in the military sanatorium "Desantnik"]. PMID- 15112412 TI - [Incidence of parasitosis in the troops of Northern-Caucasian military district]. AB - In 2002 the analysis of parasitosis incidence in the servicemen of North Caucasian military district was conducted in order to evaluate the risk of its spread and to optimize the ways of its prevention at the territory studied. One of the most important directions of prophylactic work is the training of specialists (parasitologists, laboratory physicians, epidemiologists, laboratory assistants) in the field of parasitology and early diagnosis of parasitogenic diseases in servicemen. PMID- 15112413 TI - [Ergonomic bases of improving the quality of military medical equipment]. AB - The definitions for the concepts of quality of military medical equipment (MME) sample and quality indices are given. The general ergonomic indices of the sample quality are presented. The heterogeneity of elements of "medical specialist- military medical equipment--the wounded--environment" system is shown the successful synthesis of which requires the co-ordination of technical, constructive factors, characteristics of the sample and human (personal) factors of medical specialist. The ergonomic defects typical of MME samples are revealed and the ways to improve the ergonomic characteristics of MME samples are charted. Some ergonomic aspects to improve the MME sample quality are described. PMID- 15112414 TI - [New standards for the supply of military formations with medical equipment and materials]. PMID- 15112415 TI - [The jubilee of the General Frontier Clinical Military Hospital]. PMID- 15112416 TI - [Women at the front line and the rear (From the memoirs of the front line surgeon)]. PMID- 15112417 TI - Sex, politics, and religion: why AIDS is a secular issue. PMID- 15112419 TI - Common dermatologic manifestations of herpesviruses in HIV/AIDS. PMID- 15112418 TI - Jewish religious ethics mandate access to antiretroviral drugs in developing countries. AB - The plight of millions of HIV-infected individuals without access to antiretroviral (ARV) medications constitutes an enormous problem. Religious values can influence policymakers in public and personal health issues. This article posits that Jewish religious law mandates the broadest possible access to ARV medications for HIV-infected individuals, and argues that wealthy countries must assist poorer ones to facilitate access. PMID- 15112420 TI - Ensuring quality nursing care: legal and regulatory processes. PMID- 15112421 TI - Ambulatory surgery centers--current legal issues 2004 (Part 1). PMID- 15112422 TI - Perspectives. Imagine health care swallowing 50 percent of GDP. PMID- 15112423 TI - Hysterectomy, 1981/82 to 1996/97. AB - OBJECTIVES: This article examines national and provincial trends in hysterectomies from 1981/82 to 1996/97 among women aged 35 or older. DATA SOURCES: Data for 1981/82 to 1994/95 were obtained from the Hospital Morbidity File maintained by Statistics Canada; for 1995/96 and 1996/97, from the Canadian Institute for Health Information. Supplementary data are from the 1998/99 National Population Health Survey. ANALYTICAL TECHNIQUES: Descriptive analyses present hospitalization rates for hysterectomy, the percentage performed vaginally, and average length of stay. A hierarchy of indications was used to establish the main reason for hysterectomy. Confidence intervals were calculated to determine significant changes over time and between provinces and the national level. MAIN RESULTS: From 1981/82 to 1996/97, the hysterectomy rate declined; the proportion performed vaginally increased; and average length of stay for a hysterectomy decreased. These trends generally characterized each province, although there were substantial provincial differences in rates, procedures, and average length of stay. PMID- 15112424 TI - Migraine. AB - OBJECTIVES: This article provides prevalence and incidence estimates of migraine among Canadians aged 12 or older. Associations with selected socio-demographic factors and health characteristics are also examined. Selected health indicators and medication use, as well as health care use and attitudes, are discussed, comparing migraineurs with non-migraineurs. DATA SOURCES: The findings are based on the cross-sectional and longitudinal household components of the first three cycles (1994/95, 1996/97 and 1998/99) of Statistics Canada's National Population Health Survey. Information on hospital stays is from the 1997/98 Hospital Morbidity Database, maintained by the Canadian Institute for Health Information. ANALYTICAL TECHNIQUES: Cross-tabulations were used to estimate the prevalence and incidence of migraine. Associations of migraine with selected factors were examined using generalized logistic regression. MAIN RESULTS: In 1998/99, migraine was most prevalent among women, 25- to 54-year-olds, Whites, and individuals in low-income households. The odds of being diagnosed with migraine were higher for women with pre-existing sinusitis, bronchitis or emphysema, compared with women without these conditions. The odds of this disorder for men were associated with previously diagnosed arthritis or rheumatism. PMID- 15112425 TI - Trends in colorectal cancer incidence and mortality. AB - OBJECTIVES: This article examines recent trends in the incidence of and mortality from colorectal cancer among Canadian men and women, then further analyzes trends by three subsites. DATA SOURCES: Incidence data for colorectal cancer were obtained from the National Cancer Incidence Reporting System and from the Canadian Cancer Registry. Mortality data were extracted from the Canadian Vital Statistics Database. Supplementary data on nutrition are from the National Population Health Survey. ANALYTICAL TECHNIQUES: Age-standardized incidence and mortality rates were calculated for men and women. Age-specific incidence and mortality rates were calculated by 10-year age groups. Joinpoint analysis was applied to detect statistically significant changes in linear trends. MAIN RESULTS: Since the mid-1980s, colorectal cancer incidence has been declining, with steeper rates of decrease among women. Decreasing rates of colorectal cancer are limited to tumours located in the distal colon and rectum; the incidence of cancers of the proximal colon has not changed over time. PMID- 15112426 TI - [Morphology of epidermoid carcinoma A431 cells spread on immobilized ligands]. AB - Cell interaction with extracellular matrix is a multi-step process characterized by cell attachment to substrata with subsequent cell spreading accompanied by actin cytoskeleton and cellular membrane receptor reorganization. It has been shown elsewhere that epidermoid carcinoma A431 cells, spread on solid substrata coated with fibronectin, laminin-2/4 or antibodies to EGF receptor, form specific actin filament structures typical for each particular ligand. Here quantitative analysis of heterogeneous A431 cell population spread on the above ligands has been reported. Cells were subdivided into morphological classes, according to their shape and actin filament structure, and the relationship among classes under various experimental conditions were quantitatively estimated for every ligand. We studied the influence of cell detachment pattern, short-term and long term starvation, and cell incubation in suspended state in the medium before plating on the cell population composition. It was possible to recognize the modal morphological class of cells with typical actin cytoskeleton structure dominating for the ligand in the population. Long-term starvation and incubation in suspension before cell spreading are considered as the crucial experimental parameters leading to dramatic changes in cell population. PMID- 15112427 TI - [Two types of store-operated channels in A431 cells]. AB - Activation of phospholipase C-coupled receptors leads to the release of Ca2+ from Ca2+ stores, and subsequent activation of store-operated cation (SOC) channels, promoting sustained Ca2+ influx. The most studied SOC channels are CRAC ("calcium release activated calcium") channels exhibiting a very high selectivity for Ca2+. However, there are many SOC channels permeable for Ca2+ but having a lower selectivity. And while Ca2+ influx is important for many biological processes, little is known about the types of SOC channels and mechanisms of SOC channel activation. Previously, we described store-operated Imin channels in A431 cells. Here, by whole-cell recordings, we demonstrated that the store depletion activates two types of current in A431 cells--highly selective for divalent cations (presumably, ICRAC), and moderately selective (ISOC supported by Imin channels). These currents can be registered separately and have different developing time and amplitude. Coexisting of two different types of SOC channels in A431 cells seems to facilitate the control of intracellular Ca(2+)-dependent processes. PMID- 15112428 TI - [PI 3-kinase activity is necessary for F9 mouse embryonic carcinoma cell proliferation]. AB - Embryonic carcinoma (EC) cells and embryonic stem (ES) cells have short cell cycles and, accordingly, proliferate very fast. Serum starvation does not suppress proliferation of EC and ES cells that allows to assume independence of their proliferation from the activity of cascades induced by serum. In the present work, we used flow cytometry to investigate how specific MAP-kinase and PI3-kinase inhibitors may influence proliferation and cell cycle of EC F9 cells. It is established that inhibitors of ERK-, JNK- and p38-kinases do not suppress EC F9 cell proliferation. It is possible to assume that proliferation of EC cells is supported by constitutive activity of down-stream cell cycle regulators, for example, E2F1 transcription factor. Since PI3-kinase inhibitor LY294002 causes reduction of S-phase and accumulation of G1-phase F9 cells, PI3-kinase mediated cascades seem to be constantly activated and involved in phosphorylation of important cell cycle regulators. The analysis of transcription of immediate-early genes in undifferentiated cells has shown that c-fos and c-jun genes are strongly activated by serum, and that ERK-kinase plays the main role in activation of c fos transcription, while activation of c-jun transcription depends predominantly on p38-kinase. It is necessary to note that PI3-kinase inhibitor increases effect of serum stimulation of c-fos promoter. It means that the PI3-kinase dependent cascade negatively influences the cascade, which activates c-fos transcription. Thus, the transcription of c-fos and c-jun is not connected with of EC F9 cell proliferation. The proliferation of these cells depends on PI3-kinase activity. PMID- 15112429 TI - [Wheat germ agglutinin participates in regulation of cell division in apical root meristem of wheat seedlings]. AB - A study was made of cell division activity and hormonal status in roots of 4 day old wheat seedling treated with wheat germ agglutinin (WGA). The revealed stimulating effect of WGA on mitotic index (MI) and cell area in root extension zone was specific for this lectin, because gliadin, taken as a control protein, caused no changes in growth parameters. Phytolectins (phytohemagglutinin and concanavalin A) possessing properties of mitogens rendered no such essential influence on cell growth of wheat. Immunoassay has shown that WGA-treatment leads to accumulation of auxins and cytokinins in roots. This suggest participation of WGA in regulation of MI of meristem cells in roots of seedlings during their interaction with phytohormones. PMID- 15112430 TI - [Effect of nitric oxide donor (NaNO2) on the stability of non-transformed and malignant cells to ultraviolet and gamma-radiation]. AB - A decreased sensitivity of the Chinese hamster cells (line V-79) to gamma radiation under the influence of nitric oxide induction was shown elsewhere. This effect is connected hypothetically with post-radiation reparation of DNA. The investigation of the nitric oxide donor effect on sensitivity of these to UV radiation is of interest, because this radiation is an important ecological factor of the environment. The question of retention of nitric oxide positive effect on UV and gamma-radiation sensitivity in malignant HeLa cells is no less actual, because these cells significantly differ from normal cells of line V-79. We demonstrated that the donor of nitric oxide enhances stability of the Chinese hamster cells (line V-79) to UV-radiation, as well as to gamma-radiation independently of the time of cell incubation with sodium oxide donor before or after irradiation. The inefficiency of nitric oxide as a factor increasing UV stability of cells was shown for malignant HeLa cells. A 1 h long incubation of these cells with NO-donor before gamma-irradiation decreased the number of chromosome aberrations, and conversely, the addition of this agent to the HeLa cell culture after gamma-irradiation did not change the radiostability. It may be inferred that distinctions in behaviour of nitric oxide in cultures of V-79 and HeLa cells using UV-radiation may be explained by transformation of the latter special features of their damage, and by the following reparation. PMID- 15112431 TI - [Dynamics of some postreplication DNA repair proteins in carcinogen-damaged mammalian cells]. AB - Many types of DNA lesions in template strands block DNA replication and lead to a stalling of replication forks. This block can be overcome (bypassed) by special DNA polymerases (for example, DNA polymerase eta, Pol eta) that perform translesion synthesis on damaged template DNA. The phenomenon of completing DNA replication, while DNA lesions remain in the template strands, has been named post-replication repair (PRR). In yeast Saccharomyces cerevisiae, PRR includes mutagenic and error-free pathways under the regulation of the RAD6/RAD18 complex, which induces ubiquitylation of PCNA. In mammalian cells, Pol eta accumulates in replication foci but the mechanism of this accumulation is not known. Pol eta possesses a conserved PCNA binding motif at the C terminal and phosphorylation of this motif might be essential for its interaction with PCNA. We have shown previously that staurosporine, an inhibitor of protein kinases, inhibits PRR in human cells. In this study we examined whether the accumulation of Pol eta in replication foci after DNA damage is dependent on phosphorylation of the PCNA binding motif. We also studied DNA damage-induced phosphorylation of GFP-tagged human Rad18 (hRad18) and its accumulation in replication foci. Our data indicate that (1) Pol eta is not phosphorylated in response to UV irradiation or MMS treatment, but its diffusional mobility is slightly decreased, and (2) hRad18 accumulates in MMS-treated cells, and considerable amount of the protein co localizes with detergent insoluble PCNA in replication foci; these responses are sensitive to staurosporine. Our data suggest that hRad18 phosphorylation is the staurosporine-sensitive PRR step. PMID- 15112432 TI - Bovine satellite DNA induces heterochromatinization of host chromosomal DNA in cells of trassatellite mouse embryonal carcinoma. AB - Embryonal teratocarcinoma F9 cells were transfected with a fragment (3.8 kb) of bovine satellite DNA IV (Sat), which is not homologous to mouse satellite DNA. FISH analysis revealed various chromosomal integration sites of integrated Sat in different transsatellite clones. After several passages, transsatellite had a tendency to spread along chromosome bearing Sat in one of the studied lines. The integrated transsatellites were enriched with prolonged single-strand DNA regions (SSR) revealed by FISH without previous chromosomal denaturation, and were unmethylated. The observed SSR are presumably supposed to represent intermediates of transsatellite DNA instability via unequal sister chromatid exchanges. DAPI staining demonstrated that the integrated Sat induced the formation of prominent ectopic neoheterochromatin blocks in regions adjacent to integrated Sat. These blocks were located exclusively between integrated Sat and centromeric heterochromatin. Thus, mouse repetitive centromeric DNA (AT-rich, DAPI-positive) "spreads" along the chromosome in response to integration of the bovine satellite GC-rich DNA. The results obtained are discussed in the context of possible position effect variegation mechanisms operating in undifferentiated cells. PMID- 15112433 TI - [Immortalized cell lines from murine embryos are characterized by progressive destabilization of their karyotype structure]. AB - The karyotype structure was studied for three cell lines obtained from cells of transgenic murine embryos at early stages of their establishment. The first line was obtained from a transgenic embryonic explantant containing oncogen v-sis under promotor MMTV, two other lines originated from cells of transgenic embryos containing oncogen k51. The karyotypic analysis of G-banded metaphase chromosomes revealed deviations from the normal mouse karyotype as early as by the third passage of cultivation of independent embryonic cell lines that contained a foreign oncogene in their genome. The repeated analysis that involved 15-22 passages revealed similar abnormalities: variability and progression in chromosome number with the appearance of hyperpolyploid combinations, and a large number of rearranged chromosomes, both marker and unique ones. It is concluded that introduction of a foreign oncogene into murine cell genome leads to its enhanced and progressive non-specific destabilization. Oncogen v-sis produces a more valuable karyotype destabilization than oncogen k51. PMID- 15112435 TI - [Karyological studies on Picea ajanensis (Lindl. et Gord.) Fisch. ex Carr. from different regions]. AB - Results of karyological study of Picea ajanensis (Lindl. et Gord.) Fisch. ex Carr examined from 13 provenances are presented. In addition to the cytotypes with typical chromosome number (2n = 24), P. ajanensis displays cytotypes with one or two B-chromosomes (2n = 24 + 1 - 2B). Among A-chromosomes, there are 8 pairs of long metacentrics and 4 pairs of shorter meta- or submetacentrics. Among B chromosomes there are two types of chromosomes: metacentric (B1) and submetacentric (B2) ones. There are many nucleolar chromosomes. Several chromosomes have secondary constrictions. Patterns of B-chromosome distribution within P. ajanensis are have been discussed. PMID- 15112434 TI - [Impact of angiotensin-converting enzyme, angiotensinogen, endothelial NO synthase, and bradykinin receptor B2 gene polymorphisms on myocardium in patients with hypertension and in athletes]. AB - We investigated the role of gene polymorphisms in angiotensin-converting enzyme (ACE), angiotensinogen, endothelial NO (eNO) synthase, and bradykinin receptor B2 in determining the cardiovascular system structure and function in hypertension and "athletic heart" syndrome. Using a PCR-based method, 114 hypertensive patients and 94 athletes were genotyped for I/D polymorphism of ACE, M235T angiotensinogen (ANG), Glu298 Asp endothelial synthase (eNOS), and type 2 receptor for bradykinin (BDKR2). Echocardiography and a 24 hour blood pressure monitoring being performed. The (+)-allel of BDKR2 gene was associated with the left ventricular hypertrophy and greater wall thickness in athletes and hypertensive subjects. The hypertensive patients, that were homozygous for Glu298 allele of eNOS, demonstrated a lower level of diastolic blood pressure than did those with Glu298 Asp and Asp298 Asp genotypes. At the same time, the ACE and AND gene polymorphisms displayed no association with the cardiac structure and function. PMID- 15112436 TI - [Rheological and biopharmaceutical investigation of polymer drug delivery systems]. AB - Drug release of watersoluble active agent (ephedrine hydrochloride) and water insoluble ones (griseofulvine, salicylic acid, sulfadimidine) from hydrogels containing different polymers was investigated. The rheological behaviour of polymer matrices was characterized by flow curves and viscosity curves. These hydrogels were not thixotropic systems, unlike more authors' statement. The values of rheological constants increase exponentially with concentration of polymers. The liberation of active agents can be characterized by a multiplicative function. The constant of viscosity vs. concentration function was determined and it was called as characteristic of structure formation. A linear relationship between constant of structure formation and drug liberation was demonstrated. PMID- 15112437 TI - [Importance of interfacial characteristics in pharmaceutical technology]. AB - Since drug release from the dosage forms has priority to absorption from the gastrointestinal system, physico-chemical characterisation of pharmaceutical systems is essential during the development of an optimal formulation with high efficacy and quality. Interfacial parameters of several pharmaceutical excipients were studied regarding their possible modifying effect on drug release from the dosage form. These inactive ingredients may influence the interfacial phenomena of the drug carrier system, which behaviour determines both the efficacy and the quality of the pharmaceutical preparation In this work authors deal mainly with the two introducing steps of the LADME model influenced by interfacial parameters on them, namely with the liberation of drug from the dosage form and with the characteristics influencing the absorption through biological membranes, respectively. The objective of the present work was to study modifying effects of excipients on drug liberation in connection with their physical and chemical characteristics such as interfacial tension of solid and liquid phases, wetting contact angle of solid phase and--a calculated quantity,--adhesion tension of the solid particles. PMID- 15112438 TI - [Metalloporphyrin catalyzed biomimetic oxidation potentials: potential uses and applications]. AB - Oxidative processes catalysed by metalloporphyrins nowadays are gaining importance in the study of various biological systems. One of the main direction of the investigations with synthetic metalloporphyrin catalysts is the development of systems which are capable of reproducing oxidative processes mediated by hem proteins (such as cytochrome P450, lignin peroxidase, nitric oxide synthesize (NOS), etc.). Another possible application is to develop systems containing metalloporphyrin catalyst, which could selectively oxidize organic compounds or detoxicate polluting agents at industrial level. We proved the benefits of the application of pentafluoride iron porphyrin used in three fields: -in the insect selective cytochrome P450 catalysed metabolism of carbamate insecticides [1],--to verify the natural decomposition path of insect selective cytochrome P450 inhibitors (metabolic pathways of lignin peroxidase) [2],--in modelling the H2O2 dependent NOS catalysed oxidation of the N-hydroxyguanidine based NO donors [3]. PMID- 15112439 TI - [Fragment-based drug design using stereoisomers. A case study of analogues of the phenol group in the Bioster database]. AB - This case study examined various structural features of the 55 bioisosteric fragments of the phenol group registered in version 2002.1 of the Bioster database. The size, calculated lipophilicity and H-bond donor or acceptor character of the fragments were found to vary on a fairly wide scale. In most cases, molecular modelling calculations indicated similarities in the electrostatic potential maps of the fragments. PMID- 15112440 TI - [Chiroptical study of O-heterocyclic compounds, secondary mono-alcohols and primary monoamines]. AB - Starting from cholesterol (1), model compounds containing 2,3 dihydrobenzo[b]furan or 4H-benzopyran chromophores of known configurations and rigid conformations (6, 10 and 7) were synthesized in a stereocontrolled sequence. A helicity rule was established for the 2,3-dihydrobenzo[b]furan chromophore (P/M helicity of the heteroring gives rise to negative/positive 1Lb band CD) on the basis of which the configurational assignment of norneolignans 11 13 isolated from Krameria cystisoides were revised. The synthesis and configurational assignment of the tetrahydro-2,6-methano-1-benzoxocine derivative (-)-14, homochiral with the cholestane derivative 7, was performed which allowed the study of the contribution of the chiral third and fourth sphere to the 1Lb band CD of the 4H-benzopyran chromophore. With the use of zinc porphyrin tweezer 20, an excition chirality CD method was introduced for the configurational assignment of secondary monoalcohols and primary monoamines and its applicability was proved by the study of compounds with known configuration. PMID- 15112441 TI - [Measure of alkalinity in an alcohol/water mixture by potentiometric end-point detection. Critical remarks on a new method in the European pharmacopoeia]. AB - The European Pharmacopoeia introduced a new alkalimetric method for assay of alkaloid (and other N-base) salts. The principle of method is: after preliminary addition of small amount of hydrochloric acid, titration is carried out with 0.1 N NaOH in alcohol/water mixture with potentiometric end-point detection. The applicability and reproducibility of this method have been examined at 11 substances. The results of alkalimetric method were compared to those obtained by nonaqueous titration (Ph. Hg. VII.). The method of Ph. Eur. is well reproducible (SD < +/- 0.50), however it could not be applied in case of some compounds (4 cases out of 11). Authors revealed that for the titration carried out according to the present prescription of Ph. Eur. 4., minimum pKa 7 should be the basicity of the alkaloids. They suggest to perform the titration in 70% alcohol and without preliminary addition of HCl for weaker bases. PMID- 15112442 TI - [Application of difference circular dichroism (CD) spectroscopy for determination of pharmaceutical compounds. Analysis of stereoisomers by the HPLC-CD/UV method]. AB - Circular dichroism (CD) spectroscopy is gaining an increasing importance in pharmaceutical analysis. One of the main advantages of the CD spectroscopic method is its inherent selectivity, which provides the selective analysis of chiral molecules possessing optically active absorbance bands. In addition to direct CD spectroscopic methods, several new procedures have been developed based on CD spectroscopy for the analysis and determination of pharmaceutical compounds in the past few years. One of the most progressive fields of chiroptical analysis is the combination of high performance liquid chromatographic separation and CD spectroscopy and the application of simultaneous dual CD/UV detection. The aim of our research work was to work out new procedures based on CD and CD related techniques and to investigate the application of these methods in pharmaceutical analysis. This work presents a new, selective and sensitive difference CD spectroscopic method based on oxime formation of the keto group for the determination of delta 4-3-ketosteroids and 6-keto-morphinans. Molar CD and UV spectral parameters of E and Z oxime isomers, produced in the course of oxime formation reaction, have been determined by HPLC separation and simultaneous dual CD and UV detection, without isolation of the isomers. The stereochemistry of spirocyclic pyridopyridazine compounds has been investigated by simultaneous dual CD and UV spectroscopy following HPLC separation. The enantiomeric relationship of the pyridopyridazine stereoisomers has been proved by the application of ellipticity-absorbance ratio spectra in a novel way. PMID- 15112443 TI - [Heroin, part III: the pharmacology of heroin]. AB - The major pharmacological effects of heroin can be traced back to some structural properties of the morphine molecule. The analgesic effects of heroin derive from the two active metabolites, 6-O-acetylmorphine and morphine, which bind specifically to the mu-opioid receptors of the central nervous system. mu receptors also mediate other pharmacological actions of heroin i.e. respiratory depression, euphoria and physical dependence. Heroin is more potent and faster acting than morphine as an analgesic drug. Presently, there is a considerable dispute whether heroin should be legalized for the treatment of pain in terminal cancer. Chronic administration of heroin results in the development of tolerance. It is characterized by a shortened duration and decreased intensity of the analgesic, euphoric, sedative and other CNS-depressant effects. Tolerance to opioids is due to increased adaptation of the cells which changes their receptor sites after chronic exposure to the drug. Toxicity of heroin depends markedly on the route of administration in animal studies. Variable composition of street heroin or sudden loss of tolerance can cause fatal heroin overdose of addicts. Respiratory arrest is the most common reason for death among heroin addicts. PMID- 15112444 TI - [Primary central nervous system lymphoma: diagnosis, management and current topics]. PMID- 15112445 TI - [Cigarette smoking and neurological disease]. PMID- 15112446 TI - [The influence of craniotomy on cytokines and immunological function]. AB - The purpose of the study is to elucidate the effects of a craniotomy on patients regarding the cytokines and immunological function. TNF-alpha, IL-6, the CD4/8 ratio, cortisol, and WBC were all measured perioperatively in 29 patients. No serum TNF-alpha was detectable in any patients. The IL-6 level, cortisol and neutrophils all showed an increase with a peak at three hours after the end of operation. On the other hand, the lymphocytes and the CD4/8 ratio demonstrated a opposite pattern with a negative peak at the same time, however, these levels all gradually recovered to their preoperative states on the 7th operative day. The overall effect of a craniotomy was therefore found to correlate with the length of the operation. PMID- 15112447 TI - [Garcin syndrome in a patient with rhinocerebral mucormycosis]. AB - Garcin syndrome is characterized by an unilateral cranial nerves involvement without sensory or motor long-tract disturbances. It is usually caused by tumor infiltrating in the skull base with osteolytic changes on radiological study. We report a case of 64-year-old man with history of alcohol overintake, who admitted local hospital, because of right periorbital edema and facial swelling. He noted right ptosis 2 weeks prior to admission. Neurological examination revealed right multiple cranial nerves involvement including II, III, IV, V, and VI cranial nerves. MR imaging of the brain showed marked paranasal sinusitis and abnormal infiltration of right orbital fat. Orbital apex syndrome related to paranasal sinusitis was diagnosed, and antibiotics was administered. But a few days after admission, he developed a right VII, IX, X cranial nerve palsy. He was transferred to our hospital because of acute development of left hemiparesis and deteriorated consciousness. MR imaging of the brain showed right internal carotid artery (ICA) occlusion, and infarction in right middle cerebral artery (MCA)'s territory. The diagnostic biopsy of the paranasal sinus showed mucorales hyphae, indicating that the pathological diagnosis was mucormycosis. Despite of antibiotic therapy included of amphotericin-B administration and strict control of diabetic mellitus, his sinusitis was gradually spread. His condition progressively deteriorated, and finally died of sepsis. Post-mortem examination revealed a widespread mucor infiltration in the dura mater without skull bone invasion. This case presented with unilateral multiple cranial nerve involvements (Garcin syndrome) followed by left hemiparesis associated with rhinocerebral mucormycosis. It is suggested that mucormycosis should be considered in case of Garcin syndrome without osteolysis in the skull base. PMID- 15112448 TI - [A case of peripheral-type primitive neuroectodermal tumor arising in the dura mater at the frontal base]. AB - A 7-year-old boy was admitted to our hospital because of headache and frequent vomiting. The patient was noted to have papilloedema and mild palsy of the right abducent nerve. Magnetic resonance image (MRI) revealed a large tumor in the frontal base with tumoral hemorrhage. Angiography showed the tumor was fed by anterior meningeal arteries. At surgery, the tumor was arising in the dura mater at the frontal base, and was removed totally. Histological examination showed the tumor to be composed of small cells with uniform round nuclei and minimal cytoplasm. Immunohistochemical studies were positive for MIC-2, NSE, C-KIT, vimentin, Class III-beta tublin and glycogen, but negative for NFP, synaptophysin, chromogranin A and GFAP. MIB-1 labeling index was 40-50%. The tumor was histologically confirmed to be peripheral-type primitive neuroectodermal tumor(pPNET). Following surgery, he underwent whole brain, whole spine and local radiation therapy(30 Gy in total respectively) and received two 5 day cycles of chemotherapy, consisting of intravenous administration of cisplatin 20 mg/m2/day, etoposide 60 mg/m2/day and IFOS 900 mg/m2/day. After these therapies, follow-up radiological examination showed there was no recurrence of the tumor for 24 months. Intracranial pPNET is rare. Ewing sarcoma and pPNET(ES/pPNET) is the designation given to a family of small round cell tumor arising in bone or soft tissues. Intracranial PNETs are devided into central nervous system PNET(cPNET) and pPNET. It is necessary that intracranial PNETs are divided into two types of PNETs because of different prognosis between these tumors. MIC-2 is a specific marker for pPNET/ES family and is useful in the differential diagnosis of these two types of tumors. PMID- 15112449 TI - [MR imaging study of edema along the optic tract in patient with Rathke's cleft cyst]. AB - We report an 80-year-old woman who was admitted to our hospital with symptoms due to diabetes insipidus. Magnetic resonance (MR) imaging demonstrated a sellar/suprasellar cystic lesion with marginal enhancement and the thick pituitary stalk. The MR imaging depicted edema spreading along the optic tract on fluid-attenuated inversion recovery (FLAIR) images. Upon neurological examination at the time of admission, there were no abnormal findings affecting the field of vision or visual acuity. In endocrinological examination, the basal plasma values of pituitary hormones were within normal range except for that of prolactin, which was 47.9 ng/ml. The preoperative diagnosis was craniopharyngioma, and the intrasellar mass was partially removed by the endoscopic transnasal transsphenoidal approach. Postoperative histopathological examination revealed Rathke's cleft cyst associated with squamous metaplasia. Lymphocytic infiltration was also confirmed in both the anterior and posterior pituitary lobe. The postoperative course was satisfactory. Edema spreading along the optic tract was reported as a characteristic MR imaging finding for diagnosis of craniopharyngiomas or optic nerve glioma. However, it is suggested that edema of the optic pathway seems to be caused not only by craniopharyngioma but also other suprasellar lesions. It was a rare case of secondary lymphocytic hypophysitis caused by Rathke's cleft with edema along the optic tract. PMID- 15112450 TI - [Bilateral vertebral artery dissection and its chronological changes detected by MR angiography: a case report]. AB - A case of intracranial bilateral vertebral artery (VA) dissection presenting with ischemic symptoms which following unilateral dissection is presented. A 42-year old male with an occipital headache was pointed out right vertebral artery stenosis with magnetic resonance (MR) angiography 8 day before admission. He admitted to our hospital complaining of severe vertigo and tinnitus. MR images and cerebral angiograms revealed bilateral VA dissection with infarcts in light lower surface of cerebellum perfused by posterior inferior cerebellar artery and right hypothalamus. Conservative therapy was adopted and serial MR angiography was performed. His symptoms were improved gradually and MR angiograms obtained 2 months later revealed improvement of bilateral VA stenosis. It is generally accepted that VA dissection presenting ischemic symptoms has good outcome by conservative therapy only. However, its pathological process of progression is still unknown. Based on the serial MR findings, we discuss the mechanisms of bilateral VA dissection. In this case, we consider that unilateral VA dissection extended to contralateral vertebral artery through the vertebrobasilar junction. Frequent MR angiography in acute phase could be of great use for monitoring the progression of dissection. PMID- 15112451 TI - [The posterior inferior cerebellar artery arising from the internal carotid artery directly: a variant of the persistent primitive hypoglossal artery]. AB - We reported a rare case of the posterior inferior cerebellar artery arising from the internal carotid artery directly. A 33-year-old male was admitted to our hospital with the complaint of throbbing type headache. CT showed no abnormal findings. A saccular aneurysm at the bifurcation of the left middle cerebral artery was revealed by MR angiography and the left internal carotid angiography. The right internal carotid angiography demonstrated an anomalous branch originating at the level of the C1/2 in the cervical portion of the internal carotid artery. This branch terminated as the posterior inferior cerebellar artery without an interposed segment of the vertebro-basilar artery. The ipsilateral vertebral artery was aplasia. T2-weighted MR image showed a flow-void penetrating the right hypoglossal canal. This vessel was confirmed an artery passing through the hypoglossal canal with the source images of the three dimensional time-of-flight MR angiography. We diagnosed it as a kind of variant of the persistent primitive hypoglossal artery. The persistent primitive hypoglossal artery is composed of the proximal segment derived from the primitive hypoglossal artery, and the distal segment consisting of portions of the lateral anastomotic channels (primitive lateral basillo-vertebral anastomosis) which give rise to the posterior inferior cerebellar artery. We speculated that this variant resulted from the persistence of the proximal segment, which communicated with the stem of the posterior inferior cerebellar artery via the distal segment, and next, the disconnection of the posterior inferior cerebellar artery origin with the vertebral artery due to the aplasia of right vertebral artery and the involution of the distal segment connected to the basilar artery. PMID- 15112452 TI - [Corpus callosum lipoma]. PMID- 15112453 TI - [Cystic metastatic skull tumor]. PMID- 15112454 TI - [A case of left hemiplegia following erythema, fever, and gastrointestinal bleeding]. PMID- 15112455 TI - [An experimental study on muscle tension reaction induced by voice among sign language interpreters]. AB - The purpose of this study is to ascertain the incidence of cervicobrachial muscle tension induced by voice among sign language interpreters (SLIs). The analyzed subjects included 8 professional SLIs (Case group) who complained of pain in the cervicobrachial muscle when listening to others' talking and 8 other people who have not learned sign language (Control group). Matching sexes, ages, and smoking habits of the Case and the Control groups was carried out. Informed consent to participate in the study was obtained from all subjects. While subjects relaxed in a sitting position, surface electromyographies (EMGs) were recorded from the right and left trapezius muscles and biceps brachii muscles. During this time, all subjects had two listening tasks: to listen to lectures in Japanese and to listen to instrumental music. An inquiry about subjective symptoms was conducted just after each task. Root-mean-square (RMS) values per 100 ms in each of the tasks were calculated for analysis of surface EMG. An original criterion of judgment was that the threshold of RMS values was 3.8 microV, and its threshold continued for more than 1 s. The criterion was used and the muscle tension was confirmed. Results showed muscle tension in the trapezius muscles or the biceps brachii muscles among 5 of 8 subjects in the Case group, and in 1 of 8 in the Control group when listening to the lecture. 3 subjects in the Case group and none in the Control group showed muscle tension in the trapezius muscles when listening not to music but to the lecture. The differences between the Case and Control groups in these results were not statistically significant. Confirmed muscle tension in SLIs when listening to a Japanese lecture is probably induced by the Japanese voice. It is necessary to further investigate whether muscle tension is an acquired reaction from sign language interpreting or a pathological reaction. Listening to Japanese voices may contribute to muscular overload in SLIs so that it may be necessary for SLIs to relax their muscles during break times with an absence of Japanese voices. PMID- 15112456 TI - [Method for installing an effective smoking room and the effectiveness of real time monitoring]. AB - We investigated the methodology for installing effective smoking rooms in workplaces. It is absolutely necessary to install exhaust ventilation in smoking rooms. There are two bases for deciding the exhaust ventilation rate. The most important is to eliminate the leakage of environmental tobacco smoke (ETS) from the smoking room. An airflow rate of more than 0.2 m/s at the opening of the smoking room is required by the Guidelines for Smoking Control in Workplaces (Ministry of Labour, Welfare and Health) to eliminate the leakage. This ventilation rate is decided by multiplying the opening area by 0.2 m/s. The second important point is to keep the concentration of ETS in the smoking room less than control concentration (0.15 mg/m3). This ventilation rate is decided by dividing the rate of generation of ETS by the control concentration. It is confirmed that an effective smoking room can be installed by following these guidelines. We used real-time monitoring to evaluate the leakage of ETS from the smoking room and the ETS concentration in the smoking room before and after the improvement. It is concluded that real-time monitoring of ETS is a useful method for evaluating the effectiveness of the smoking room. PMID- 15112457 TI - [Research toward direct on-filter IR analysis of silica dust by means of a rotating filter holder]. PMID- 15112458 TI - Current and future therapeutic opportunities in the comorbidity between the epilepsies and affective disorders. PMID- 15112459 TI - Current status of the utilization of antiepileptic treatments in mood, anxiety and aggression: drugs and devices. AB - Interventions that have been utilized to control seizures in people with epilepsy have been employed by the psychiatric community to treat a variety of disorders. The purpose of this review will be to give an overview of the most prominent uses of antiepileptic drugs (AEDs) and devices like the Vagus Nerve Stimulator (VNS) and Transcranial Magnetic Stimulation (TMS) in the treatment of psychiatric disease states. By far, the most prevalent use of these interventions is in the treatment of mood disorders. AEDs have become a mainstay in the effective treatment of Bipolar Affective Disorder (BAD). The U.S. Food and Drug Administration has approved the use of valproic acid for acute mania, and lamotrigine for BAD maintenance therapy. AEDs are also effectively employed in the treatment of anxiety and aggressive disorders. Finally, VNS and TMS are emerging as possibly useful tools in the treatment of more refractory depressive illness. PMID- 15112460 TI - Convergences in course of illness and treatments of the epilepsies and recurrent affective disorders. AB - The failure to achieve and maintain remission is a critical problem for a high percentage of patients with epilepsy and the primary affective disorders. Early illness onset and delayed initiation of treatment may contribute to primary treatment resistance or that associated with loss of efficacy (tolerance phenomenon). Neurobiological data and principles drawn from the amygdala kinding model of seizure progression are reviewed for their heuristic value in conceptualizing molecular mechanisms of illness progression and its prevention with pharmacological agents in the epilepsies and, indirectly, the recurrent affective disorders. Caveats in the use of this model and convergences and divergences in its predictive validity for seizures and affective disorders are noted. PMID- 15112461 TI - Shared mechanisms of antidepressant and antiepileptic treatments: drugs and devices. AB - Many treatments for the epilepsies and affective disorder share the properties of seizure suppression and mood stabilization. Moreover, affective disorders and the epilepsies appear to share partially similar pathogenic mechanisms. A component of the shared predisposition appears to arise from noradrenergic and serotonergic deficits. Increasing evidence supports the hypothesis that noradrenergic and/or serotonergic elevation is a mechanism of therapeutic benefit shared by most antidepressants and many antiepileptic medications. Medication induced alterations in GABAergic, glutamatergic, and CRH (corticotropin releasing hormone) containing neurons may also contribute to the shared therapeutic properties of antidepressant and antiepileptic medications. PMID- 15112462 TI - Epilepsy and depression: imaging potential common factors. AB - Patients with seizure disorders have an increased incidence of depression. This may be due in part to psychosocial factors; or side effects of antiepileptic drugs. However, there may be underlying physiologic mechanisms for the relationship. Neuroimaging studies, including structural magnetic resonance imaging, positron emission tomography measurements of cerebral glucose metabolism, and, more recently, imaging of serotonin 1A receptors, may provide additional data to explain overlapping clinical manifestations of epilepsy and depression. PMID- 15112463 TI - Structural MRI changes of the brain in depression. AB - For many years, investigators have been trying to identify the neuroanatomical structures responsible for the development of neuropsychiatric disorders, specifically depression and schizophreniform disorders. The available data were based on observations made in neurological patients who developed a psychiatric comorbid disorder following the neurologic insult. With the advances in high resolution magnetic resonance imaging and functional neuroimaging studies, we have witnessed in the last decade a wealth of new data that identify structural neuroimaging changes in mesial temporal structures, prefrontal cortex and basal ganglia in major depressive disorders. The purpose of this article is to briefly review the published data on neuroanatomical structural changes associated with major depressive and bipolar disorders. PMID- 15112464 TI - Affective disorder and epilepsy comorbidity: implications for development of treatments, preventions and diagnostic approaches. AB - Concepts pertaining to affective disorder and epilepsy comorbidity are contributing appreciably to improvements in patient care. Several antiepileptic treatments have become important components of the management of bipolar affective disorder. In contrast, little progress has emerged in developing clinical applications of the anticonvulsant properties of the antidepressants in the treatment of the epilepsies. The slow onset of action of the antidepressants remains a major impediment to fully effective treatment of depressive episodes. Nevertheless, studies from experimental epileptology demonstrate that the anticonvulsant effects of the antidepressants occur rapidly and as a consequence of noradrenergic and/or serotonergic activation. These studies also demonstrate that adequate initial doses of the antidepressants are essential to rapid onset of anticonvulsant action. Pharmacokinetically valid loading dose paradigms are seemingly avoided with antidepressant drugs in humans because of potential toxicities and/or patient unacceptability. However, substantial progress has been made in reducing the adverse effect liability of the antidepressants. No longer is convulsive liability considered to stem from the therapeutic mechanisms of the anti-depressants. Rather, noradrenergic and serotonergic influences have demonstrable anticonvulsant properties. Other side effects may also be separable from the anticonvulsant and antidepressive effects of antidepressive treatments. The concept that the protracted process of antidepressant-induced beta noradrenergic down-regulation is an essential prelude to the onset of mood benefit is no longer a sustainable premise. Nevertheless, increasing evidence underlies the possibility that knowledge of serotonergic and noradrenergic regulatory processes can be used to design strategies that will hasten the onset of antidepressive action. Similar optimism pervades efforts to determine the possibility that dual inhibition of serotonin and norepinephrine transporters will hasten onset of antidepressive action. Moreover, because noradrenergic and serotonergic systems are determinants of predisposition to seizures and to dysfunctional affective episodes, augmentation strategies may also be applicable to the use of antidepressant drugs in epilepsy and to the use of antiepileptic drugs such as carbamazepine in mood disorders. Recent studies have demonstrated that, in part, the therapeutic effectiveness of carbamazepine may stem from its marked capacity to elevate serotonin concentrations in the extracellular fluid of the brain via mechanisms that differ from those of the membrane reuptake inhibitors. Evidence suggests that the epilepsies and affective disorders may arise from a multiplicity of neurobiological abnormalities. A disorder in one individual may arise via different mechanisms than a phenomenologically similar disorder in another individual. Thus, diagnostic tools are needed to make mechanistic distinctions among individuals so that treatments can be appropriately developed and selected. In terms of epileptogenesis and affective disorder progression, neuroprotective paradigms for one individual may differ from those needed for another. Moreover, diagnostic technologies that are adequate to detect genetically and/or experientially determined vulnerability before the onset of a seizure or dysfunctional affective episode may be valuable steps toward achieving goals of prevention. PMID- 15112465 TI - [Polish Gynecological Society's recommendations regarding contraception]. PMID- 15112466 TI - [Lipid peroxides concentration in women with intrauterine growth restriction]. AB - OBJECTIVES: Intrauterine growth restriction is connected to high free radicals activity. One of the indices of lipid cell membrane destruction in free radical processes are lipid peroxides. DESIGN: The aim of the study was to find correlation between lipid peroxides products concentration in healthy women and intrauterine growth restriction. MATERIAL AND METHODS: The study was done in Medical University in Lodz in 1999-2002. The study group consist of women with IUGR, the controls of healthy pregnant women. The lipid peroxides were measured by colometric method, using Bioxytech kit for lipid peroxides. RESULTS: In IUGR group concentration of lipid peroxides was 42.17 +/- 11.77 mumol/l. In controls 53.754 +/- 14.07 mumol/l. We found significant difference between two groups and the increase of lipid peroxides in IUGR group. CONCLUSIONS: In women with IUGR the processes of lipid peroxidation increased. PMID- 15112467 TI - [Leptin concentration in women with normal pregnancy and intrauterine growth retardation]. AB - OBJECTIVES: Leptin takes part in fat regulation and can also take part in regulation of prenatal fetal weight. DESIGN: The aim of the study was to find correlation between leptin concentration in healthy women and intrauterine growth restriction. MATERIAL AND METHODS: The study was done in Medical University in Lodz in 2000-2002. The study group consisted of 50 women with IUGR, the control group of 50 healthy pregnant women. Elisa immunoenzymatic IBL test was used. The results were expressed in microgram/l. RESULTS: In group of normal pregnancy leptin concentration was 5.347 +/- 1.098 micrograms/l. In group of intrauterine growth restriction leptin concentration was 4.617 +/- 0.949 micrograms/l. The difference between groups was statistically significant but the mean values in both groups were normal for nonpregnant women. CONCLUSIONS: Low concentration of leptin can play role in intrauterine growth restriction. PMID- 15112468 TI - [Plasma carbonyl group concentration in pregnant women with IUGR treated by L arginine and acetylsalicylic acid]. AB - OBJECTIVES: Carbonyl groups are the elements connecting protein structure, they influent into biological activity. High concentration of carbonyl groups means high risk of protein destruction. DESIGN: The aim of this study was the evaluation of carbonyl group concentration in blood serum in normal pregnancy and in women with IUGR treated by L-arginine and acetylsalicylic acid. MATERIAL AND METHODS: The study was done at the Department of High Risk Pregnancy, Department of Gynaecology and Obstetrics, Medical University of Lodz, in 1999-2002. The study group included 80 pregnant women hospitalised due to foetal growth restriction, between the 32-th and 38-th weeks of pregnancy. The treatment was conducted for twenty days and consisted of everyday low dose aspirin (Acard) and L-arginine (NO precursor). The ultrasound and laboratory examinations were done on the first day of hospitalisation in both groups, and at 20-th day of treatment in the group of IUGR. The carbonyl groups concentration was measured by Levine method and expressed in mmol/1 mg proteins. RESULTS: In controls mean value of carbonyl groups concentration was 1.848 +/- 0.291 mmol/1 mg proteins and after 20 days of observation 1.897 +/- 0.439. In the group with IUGR before treatment was 2.193 +/- 0.658. After 20 day of the therapy and the value decreased to 2.078 +/- 0.679. There is a significant difference between the value of carbonyl-groups concentration in normal pregnancy and IUGR. The carbonyl-groups concentration decreased after 20 days of treatment by L-arginine, and the value is still higher than in normal pregnancy. CONCLUSIONS: Oxidative protein damage in IUGR decreased in the process of treatment. PMID- 15112469 TI - [Non-invasive fetal RhD typing and RhD negative pregnant women--preliminary observations]. AB - OBJECTIVES: For a follow up of a pregnancy in a RhD neg women it is crucial to learn whether her fetus is RhD pos or neg. AIM: Detection of fetal RHD gene in the plasma of RhD neg mother in various periods of pregnancy and a comparison with RhD of a newborn. 45 plasma samples from various periods of pregnancy from 28 RhD neg women. MATERIAL AND METHODS: Examination of: RHD (exon 7 and 10), SRY, GSTM1 and ACE using real-time PCR. RESULTS: Fetal RHD was detected in all 23 mothers with RhD pos child; in one of them, in the 12th week of gestation, RHD was detected only using primers for exon 7, however in the 21st--the presence of both RHD exons was confirmed. Five mothers delivered RhD neg newborns--both RHD exons were not detected. In all 5 cases other fetal genes were examined to be certain that the fetal DNA was analysed. In all 3 mothers who delivered RhD neg boys, the SRY was detected. Among 2 mothers who delivered RhD neg girls, the presence of fetal DNA was confirmed in one, by GSTM1 detection. CONCLUSIONS: 1) Real-time PCR is an appropriate, non-invasive method for fetal RhD examination. 2) Two RHD exons should be examined. 3) Control genes should be investigated and found to be sure that negative result of the RHD was not false negative due to the lack of fetal DNA; SRY is an appropriate control for boys, while for girls further investigations are needed, which are in progress. PMID- 15112470 TI - [Assessment of mothers' knowledge about breastfeeding]. AB - OBJECTIVES: The aim of this study was to assess the level of knowledge of problems referring to breastfeeding among women in childbirth. MATERIALS AND METHODS: The study was based on a questionnaire, which consisted of 20 questions with multiple choice answers. 108 women in puerperium were surveyed-54 in the Obstetrical Ward in Puck and the same number in the I Clinic of Obstetrics and Gynaecology in Warsaw. The results were analysed according to age, education and attendance to labour school. Statistical analysis was performed afterwards. RESULTS: For all the surveyed women, breastfeeding is a natural need and an important element of the emotional bond with a newborn. For 78.7% of the women questioned, the priority of breastfeeding was the health of their baby, 16.7% point to the advantages of the optimal composition of human milk, 37% of mothers found breastfeeding convenient. On the other hand, 37% consider artificial milk only a substitute. For 60% of women magazines were the source of education, books 53.7%, acquaintances and family 32.4%. Only a small percentage (24.1%) of women took advantage of labour classes, although it is an extremely successful method of teaching. 17.6% of women were not looking for any information about breastfeeding during the pregnancy. CONCLUSIONS: Differences in preferences and level of knowledge between women from big and small centre were shown, although there was no difference in access to it. Magazines and books, not medical staff, were the most popular source of information. The level of knowledge depends on education and labour class attendance. It seems that the system of lactational guidance should be extended on obstetrical wards, concentrating especially on women with poorest education. PMID- 15112471 TI - [Use of the Hasson technique for creating pneumoperitoneum in laparoscopic surgery]. AB - OBJECTIVE: The aim of this study was to estimate safety and efficacy of Hasson technique in laparoscopic surgery. MATERIAL AND METHOD: In those patients where adhesions are suspected pneumoperitoneum was achieved by open Hasson technique. The time of this procedure, changes of hemoglobin concentration, duration of hospitalization and postoperative pain was compared with those cases where pneumoperitoneum was performed by blind insertion of the Veress needle. RESULTS AND CONCLUSIONS: Hasson technique is more time consuming procedure but its safety and efficacy is significantly higher in those patients where adhesions are suspected. PMID- 15112472 TI - [Effect and tolerability of sequential administration of estradiol valerate and medroxyprogesterone acetate on menopausal symptoms, hormonal profile and biochemical markers in climacteric women]. AB - Hormonal replacement therapy is the gold standard in the treatment and prophylaxis of menopausal symptoms and osteoporosis. THE AIM: Of the study was to assess changes in climacteric symptoms, hormonal profile and biochemical parameters during sequential hormonal replacement therapy in climacteric women 19 women aged 46-61 years old was included into the study. We evaluated Kuppermann's index, serum concentration of estradiol, FSH, morphology of the blood, lipids and haemostatic parameters. RESULTS: The menopausal symptoms significantly decreased. Lipids levels, haemostatic parameters and morphological parameters of the blood didn't change. CONCLUSION: The sequential hormonal replacement therapy with estradiol valerate and medroxy-progesterone acetate is efficient and safe. PMID- 15112473 TI - [Von Willebrand disease in women with menorrhagia]. AB - Excessive menstrual bleeding is a common clinical problem in women of reproductive age. Hereditary bleeding disorders, such as von Willebrand disease, may be one of the causes of menorrhagia. The aim of our study was to assess the frequency of von Willebrand disease and other hemostatic defects in women with unexplained menorrhagia and without uterine pathology. Sixty-five women (mean age 31 +/- 8.7, range 18-48 years) presenting with menorrhagia were screened. An inherited bleeding disorder was diagnosed 9 (13.8%) patients; the disorders were von Willebrand disease of mild severity (type 1) in 8 cases (12.3%) and a platelet dysfunction (storage pool disease) in 1. The frequency of bleeding symptoms was higher in patients with an inherited hemostatic defect than in women without bleeding disorder. The results of this study underscore the need for hemostatic evaluation in women with excessive menstrual bleeding and without obvious pelvic abnormality. PMID- 15112474 TI - [Case report of 18 weeks delayed delivery of the second twin after the miscarriage of the first fetus at the 17th week of pregnancy]. AB - Delayed delivery of the second twin after the miscarriage of the first foetus is very rare. Delayed second-twin delivery gives the opportunity for corticoid and antibiotics administration--procedures that decrease the infant morbidity and mortality. We report a case of the second twin's retention after miscarriage of the first at 17th week of pregnancy. The delay time was of 126 days (18 weeks). Pregnancy like this is at increased risk for all maternal complications, in particular intrauterine infection. Parents consent have to be obtained after informing them about advantages and risk of such procedure. The patient was treated with tocolysis, antibiotics, corticosteroids but not previous cervix cerclage has been performed. On regards of our experience and the data from literature we conclude that when observing fetal status and maternal condition, it is possible to delay the second twin's delivery long time after the miscarriage or birth of the first one thus giving the chance to the second twin to grow more mature. The main problem is imminent infection which defines the prognosis. Cerclage does not seem to be essential for better outcome. PMID- 15112475 TI - [Ovarian cancer--therapeutic options after the failure of the first line of treatment]. AB - Ovarian cancer is the fifth leading cause of cancer deaths in women. The "gold standard" of the first line setting is taxanes and platinum-based chemotherapy. Despite substantial response, most women with ovarian cancer are destined to relapse, which occurs in 18-22 months. Management of recurrent ovarian cancer is still challenging. In this article we reviewed recent methods of treatment of the relapsed ovarian cancer: chemotherapy, radiotherapy and experimental approaches. Platines-derivates monotherapy seems to be the most adequate treatment for the platinum-sensitive ovarian cancer (carboplatin). Patients with platinum refractory ovarian cancer should be considered for oral etoposide, radiotherapy or topotecan according to the current course of the disease. Once ovarian cancer recurs, cure is no longer a realistic goal. So it is important to realize, that the benefit with improved quality of life and relieving symptoms will be the main aspect of this treatment. PMID- 15112476 TI - [History of vaginal reconstruction]. AB - The paper presents the outline of the history of vaginal reconstruction since antiquity, mainly in vaginal agenesis. The review presents first concepts of vaginoplasty originating from Hippocrates and Celsus, a few attempts of vaginal reconstruction in 18th-19th centuries nad moderate advances in the USA in 19th century. A breakthrough in this field was related to introduction of split thickness skin graft for covering neovaginal channel by Abbe at the turn of 19th and 20th century. Subsequent evolution of surgical techniques was based on Abbe's concept and was directed to finding an ideal material accelerating the epitheliazation of vaginal channel. At the turn of 20 and 21st century medical technology development e.g. laparoscopy played its role in this field of surgery too. Tissue engineering and molecular biology will definitely play increasing role in vaginal reconstruction. Conservative and minimally invasive methods of vaginal creation were also briefly discussed. The considerable input of Polish scientist into the development of both experimental and clinical problems of vaginoplasty is presented. PMID- 15112477 TI - [Report from the Conference of the Polish Gynecological Society, Poznan, December 13, 2003]. PMID- 15112478 TI - Service is put on notice over poor-quality data. PMID- 15112479 TI - Data briefing. Mortality rates. PMID- 15112480 TI - Clinical management. The late show. AB - The new GP contract will be a catalyst for nurse prescribing. With most GPs choosing to exercise their right to opt out of out-of-hours care, nurse prescribing will be a key part of establishing new multidisciplinary teams. Factors hindering progress may include nursing shortages and limitations of the independent nurse formulary. PMID- 15112481 TI - Change management. Culture vultures. PMID- 15112482 TI - Law. Danger money. PMID- 15112483 TI - Mental health. Body matters. PMID- 15112484 TI - HSJ people. Nature and nurture. PMID- 15112486 TI - [Oncogenic emergency and treatment in thyroid disease]. AB - The urgent treatment of thyroid disease is mainly considered to be that of thyroid neoplasia, especially malignant thyroid tumors. The clinical symptoms produced by malignant neoplasia are acute respiratory failure caused by tumor compression and/or invasion of the trachea. Massive bleeding caused by tumor invasion of the major vessels also requires emergency therapy. In general, thyroid cancer has a favorable prognosis in patients with well-differentiated papillary carcinoma, which makes up more than 90% of thyroid malignancies. However, anaplastic carcinoma, undifferentiated papillary carcinoma, and malignant lymphoma originating from the thyroid sometimes require urgent therapy to save lives. Anaplastic carcinoma that is basically transformed from well differentiated papillary carcinoma shows rapid growth and invades the surrounding tissue to produce tracheal obstruction. Repeated postoperative recurrences in patient with papillary carcinoma suggest undifferentiated carcinoma that eventually becomes the same endostage as anaplastic carcinoma. Radical surgery is needed in such cases initially, although making a differential diagnosis between well- and undifferentiated carcinoma is extremely difficult. Urgent treatment of these diseases consists of ensuring that the airway remains open to improve quality of life, but does not include radical treatment. Stent insertion is one adequate method for the urgent treatment of respiratory disturbance caused by tumor compression. Malignant lymphoma frequently develops from Hashimoto's thyroiditis. Immediately after making an accurate pathologic diagnosis, chemotherapy and/or radiation therapy are effective in reducing enlarged goiters rather than surgical management. PMID- 15112487 TI - [Oncogenic emergencies: appropriate treatment in lung cancer]. AB - Several critical situations in patients with lung cancer are considered oncogenic emergencies. For example. massive hemoptysis, severe airway stenosis, retention of large amounts of pleural and pericardial effusion, and superior vena cava syndrome are situations requiring immediate care. For uncontrollable massive hemoptysis, embolization of the bronchial artery is a useful treatment method. For airway stenosis, rapid widening of the airway by endoscopic surgery is necessary. Since massive retention of pleural or pericardial effusion is life threatening prompt drainage is required. For brain metastasis with increased intracranial pressure, decompression surgery is useful to prevent brain herniation. Spinal cord compression by epidural metastasis is considered to be an indication for emergency decompressive laminectomy in selected patients. Since the severity of emergent situations varies individually, the appropriate treatment should be determined based on the clinical condition of individual patients. PMID- 15112488 TI - [Treatment of oncogenic emergencies in esophageal disease]. AB - Common oncogenic emergent conditions of the esophagus are esophageal fistula with malignancy and peptic ulcer, perforation by a foreign body, and rupture (Boerhaave's syndrome) and bleeding with malignancy. The current standard of palliative therapy for patients with malignant tracheoesophageal fistula is endoscopic replacement using covered self-expandable metallic stents in the esophagus and/or trachea. We successfully treated two patients with esophageal bleeding caused by malignant ulceration. To prevent the formation of an aortoesophageal fistula, a covered self-expandable metallic stent was inserted into the esophagus and aorta. Insertion of covered self-expandable metallic stents in patients with esophageal malignancies significantly improves dysphagia, seals fistulas/perforations and ulcerations, and is associated with acceptable morbidity and mortality rates. Spontaneous esophageal rupture, also known as Boerhaave's syndrome, is a rare condition. Primary repair is appropriate for ruptures diagnosed early. Many are diagnosed late and T-tube drainage may be the simplest way to manage this difficult condition in this situation. PMID- 15112489 TI - [Oncologic emergencies in gastric cancer patients]. AB - Many complications frequently occur in gastric cancer patients which require urgent treatment. Oncologic emergencies in gastric cancer vary widely and include hemorrhage, perforation and obstruction due to gastric cancer tumors, obstructive jaundice, hydronephrosis, intestinal obstruction and disseminated intravascular coagulation due to advanced metastatic, recurrent, or systemic tumors, and adverse effects secondary to chemotherapy. In gastric cancer treatment, we must recognize the occurrence of oncologic emergencies resulting from gastric cancer progression and recurrence. It is important that the knowledge of advanced stages and the prognosis of gastric cancer patients be taken into consideration when treating patients in a critical state. PMID- 15112490 TI - [Oncologic emergencies in colorectal cancer patients]. AB - The management of obstruction and perforation, which are representative of primary oncologic emergencies in colorectal cancer patients, is outlined. In the management of obstructing colorectal carcinoma, primary resection and anastomosis can be performed in elective conditions with favorable outcomes with preoperative decompression of the obstruction using decompression tubes or self-expanding stents and is the treatment of choice. Even in patients in whom the obstruction fails to improve, the current trend favors intraoperative on table lavage followed by primary resection and anastomosis, except when patients are hemodynamically unstable during surgery or when the condition of the bowel is not optimal for primary anastomosis. For patients with colorectal perforation, the treatment of first choice is primary resection of the septic focus at the perforation site plus colostomy. Single-stage surgery (primary anastomosis after resection of the perforation site) or primary resection of the tumor in patients with perforation occurring far proximal to the cancer should be avoided to reduce the risk of postoperative complications, especially in patients with serious conditions. Intensive management to prevent sepsis including continuous blood purification methods such as polymixin B-immobilized fiber hemofiltration and continuous hemodiafiltration is required to improve the perioperative mortality rate. PMID- 15112491 TI - [Management of spontaneous ruptured hepatocellular carcinoma]. AB - Spontaneous rupture is a major life-threatening complication of hepatocellular carcinoma(HCC). Ruptured HCC often causes hypovolemic shock and hepatic hypoperfusion. Patients with impaired liver function tend to lapse into liver failure, which is the main cause of death. To prevent liver failure, accurate diagnosis and adequate treatment for the restoration of the efficient liver perfusion are required. Emergent transarterial embolization (TAE) is the most effective and less-invasive treatment for hemostasis. On the other hand, emergent hepatic resection should be avoided because of the increased risk of postoperative liver failure and incomplete resection of the tumor. After achieving hemostasis, a second-stage therapeutic approach for HCC is required. Patients with acceptable liver function should undergo surgery. The prognosis of ruptured HCC treated with second-stage hepatectomy is considered to be comparable with that of nonruptured HCC. In conclusion, spontaneous rupture of HCC is considered to be a combination of acute and severe peritoneal hemorrhage with malignant disease. To improve the prognosis, adequate early treatment for the control of hemorrhage while preserving liver function is an important factor. TAE followed by elective hepatectomy is considered the most effective treatment. PMID- 15112492 TI - [Oncologic emergencies associated with pancreatobiliary cancer]. AB - Clinically common oncologic emergencies associated with pancreatobiliary cancer are gastrointestinal bleeding caused by duodenal invasion of pancreatic carcinoma, severe duodenal obstruction due to pancreatic carcinoma, and acute cholangitis accompanied by obstructive jaundice in patients with biliary tract carcinoma. When a patient with gallbladder cancer presents with acute cholecysitis, emergency surgery is sometimes performed on the basis of the latter diagnosis. Emergency procedures can also be required in the perioperative management of pancreatobiliary cancer, for example, in biliary peritonitis caused by detachment of a percutaneous transhepatic biliary drainage (PTBD) tube and in ruptured pseudoaneurysm due to postoperative pancreatic or biliary leakage. Nonsurgical procedures are usually initially selected for oncologic emergencies associated with pancreatobiliary cancer, because patients are likely to develop severe organ dysfunction and it is difficult to access directly and remove the pancreas or biliary tract during emergency surgery. When systemic conditions improve, it is necessary to evaluate the degree of disease progression and systemic conditions, and if feasible, the primary lesion should be surgically resected. When performing emergency cholecystectomy in patients with acute cholecystitis, thorough intraoperative investigation of resected specimens is important, considering the possibility of concomitant gallbladder carcinoma, since thorough examination cannot be performed in such emergency settings. Furthermore, when cholangitis accompanies pancreatobiliary cancer, emergency drainage should be considered as sepsis can develop rapidly. PMID- 15112493 TI - [Surgical practice at a private coloproctology hospital]. PMID- 15112494 TI - [What we can learn from a case of medical malpractice]. PMID- 15112495 TI - [Protective effect of carbon monoxide for organ injury]. AB - Heme oxygenase (HO) catabolizes heme into three products: carbon monoxide (CO), biliverdin and free iron. HO-1, inducible form of HO, has been shown to be protective against various stress. CO at a low concentration has been shown to be protective in several disease models mimicking the action of HO-1. We suggest that CO, the byproduct of heme degradation, could be valuable therapeutic agents. We review the functional role of HO-1 and CO and its potential application to clinical settings. PMID- 15112496 TI - Death-defying empathy. PMID- 15112497 TI - At the interface of cultures. PMID- 15112498 TI - A defense of the philosopher-ethicist as moral expert. PMID- 15112499 TI - Moral reasoning of members of hospital ethics committees: a pilot study. PMID- 15112500 TI - Ethics consultation: in the service of practice. PMID- 15112501 TI - Peter Singer and beastiality. PMID- 15112502 TI - On judicial obstruction of sound surrogate decision making: a comment on California's Wendland case. PMID- 15112503 TI - [Change in feelings of regret over time: relation to decision-making style, behavior, and coping methods]. AB - This study investigated the change in feelings of regret over time in relation to decision-making style, critical thinking, behavior, and coping methods with the regret. Seventy undergraduate students completed a questionnaire on critical thinking, decision-making styles, feelings of regret and the coping methods in five different situations: entrance examinations, declarations of love, skiing, career changes, and investments. Results showed the following. First, in situations which normally occur only once (i.e., entrance examinations), subjects who indicated inaction felt increased regret over time, but those who indicated action felt decreased regret. Conversely, in situations which occur regularly (i.e., declarations of love and skiing), both of those who indicated action and inaction felt decreased regret. Second, people who indicated action coped with their regret using the method of rationalization more often than those who indicated inaction. In situations which normally occur only once, analytic decision-makers tended to cope with their regret by improving their behavior more than intuitive decision-makers. Finally, critical thinkers tended to adopt an analytic style more often than an intuitive style. PMID- 15112504 TI - [Explicit memory for type font of words in source monitoring and recognition tasks]. AB - We investigated whether people can consciously remember type fonts of words by methods of examining explicit memory; source-monitoring and old/new-recognition. We set matched, non-matched, and non-studied conditions between the study and the test words using two kinds of type fonts; Gothic and MARU. After studying words in one way of encoding, semantic or physical, subjects in a source-monitoring task made a three way discrimination between new words, Gothic words, and MARU words (Exp. 1). Subjects in an old/new-recognition task indicated whether test words were previously presented or not (Exp. 2). We compared the source judgments with old/new recognition data. As a result, these data showed conscious recollection for type font of words on the source monitoring task and dissociation between source monitoring and old/new recognition performance. PMID- 15112505 TI - [The validity of the three dimensional model to classify coping behavior]. AB - The purpose of this study was to validate the three dimensional model of classifying coping behavior. The three are: Encounter-Avoidance, Problem-Emotion, Behavior-Cognition Dimension. A set of questionnaires on coping behavior and psychological stress responses were administered, and 1,604 undergraduates and 1,296 adults completed them. Analyses of covariance structure were performed, and results indicated that the three dimensional model, with eight coping style, showed the highest goodness-of-fit statistics. Multiple regression analyses also indicated that Problem-Emotion and Behavior-Cognition dimensions were useful for predicting the coping effects on psychological stress responses. Therefore, the two dimensional model of Problem-Emotion and Behavior-Cognition, with four coping style, would be helpful for further investigation of coping behavior that would buffer psychological stress responses. PMID- 15112506 TI - [The effect of empathy on accuracy of behavior prediction in social exchange situation]. AB - Seventy-six participants first played a prisoner's dilemna (PD) game with an anonymous partner and then predicted behavior of other participants in the same PD game. In addition, they took a test of accuracy in judging sociometric relations among group members. Of the four factors of empathy, the two reflecting emotional aspects--self-directed and other-directed emotional responses--were found to have no relation with accuracy in predicting behavior of other participants and judging sociometric relations among group members. In contrast, the other two reflecting cognitive aspects--imagination and perspective taking- were found to be related to accuracy in predicting behavior of other participants in the PD game. Of the two, imagination was a better indicator of accuracy in the prediction than perspective taking. Implications of these findings are discussed in terms of domain specificities of adaptive task that different aspects of empathy are thought to fulfill. PMID- 15112507 TI - [The calming process of anger experience: time series changes of affects, cognitions, and behaviors]. AB - This study investigated time series changes and relationships of affects, cognitions, and behaviors immediately, a few days, and a week after anger episodes. Two hundred undergraduates (96 men, and 104 women) completed a questionnaire. The results were as follows. Anger intensely aroused immediately after anger episodes, and was rapidly calmed as time passed. Anger and depression correlated in each period, so depression was accompanied with anger experiences. The results of covariance structure analysis showed that aggressive behavior was evoked only by affects (especially anger) immediately, and was evoked only by cognitions (especially inflating) a few days after the episode. One week after the episode, aggressive behavior decreased, and was not influenced by affects and cognitions. Anger elicited all anger-expressive behaviors such as aggressive behavior, social sharing, and object-displacement, while depression accompanied with anger episodes elicited only object-displacement. PMID- 15112508 TI - [Causal attribution and conflict resolution in the Japanese language instruction setting]. AB - Conflicts sometimes arise between the language teachers and foreign students in the Japanese language instruction setting. The purpose of this study was to investigate the relationship of causal attribution and selection of conflict resolution strategies in such conflicts, and to explicate the characteristics of intercultural conflicts resolution strategies, especially paying attention to the teacher expectation of student strategy to resolve a conflict. A questionnaire was administered to 84 Japanese teachers, 214 Chinese students, and 154 Korean students. Results indicated that when students attributed the conflict to the teachers, they chose assertive strategies, and that when attributed to themselves, compromising strategies were used. Moreover, while Chinese students tended to use such compromising strategies as cooperation and obedience when they attributed the conflict to cultural differences, Korean students did not tend to use non-confrontational strategies. The teachers did not expect their students to use assertive strategies, and the large gap between teacher and student perception of assertive strategies could be one of reasons for difficulties that arise in such intercultural conflict resolution. PMID- 15112509 TI - [Recognition of vocal expression of emotion and its acoustic attributes]. AB - The vocal expression of emotion was examined with both an auditory experiment and objective acoustic analyses. In the auditory experiment the stimuli were words or short sentences with six basic emotions expressed by two actors. Forty-five undergraduate and graduate students participated in the experiment. The results showed that the vocal expression of emotion was strongly identified, except in the case of fear, and that the six basic emotions could be plotted in a psychological space with two dimensions calculated from multidimensional scaling. The plot formed a roughly circular surface, with locations very similar to those of the facial expressions. One dimension was considered to represent the element of pleasantness-unpleasantness. The actors voices were then acoustically analyzed. The results suggested that the mean fundamental frequency (F0), the standard deviation (SD) of F0, and the SD of the energy are the important factors that define the characteristics of the vocal expression of emotions. To determine the most important parameter(s) and explain the two dimensions of the psychological space, canonical correlation analysis was conducted. The analysis indicated that F0 was correlated with the pleasantness-unpleasantness dimension. PMID- 15112510 TI - [Self-reference effect in an independence/remember-know procedure]. AB - The self-reference effect was examined using the IRK (independence/remember-know, Jacoby, 1998) procedure. In the learning phase, participants were asked to rate trait words in one of the three ways of encoding: self-reference, semantic, and physical. In the test phase, they were told to use a word-stem as a cue to recall studied words, if they could not recall, to complete stems with the first word that came to mind. Furthermore, participants were to classify the completed words into those they "remembered," those they "knew," or "new." Estimates derived from the IRK procedure showed the self-reference effect only for the intentional memory (i.e., the "remembered" items), but not for the automatic memory. Results suggested that the self-reference effect is due to intentional use of memory. PMID- 15112511 TI - [A multi-dimensional approach to delusional ideation in normal people: comparison with paranoid ideation and guarded ideation]. AB - Recent studies reported that normal people have delusional ideation. The purpose of the present study is to compare the types of delusional ideation in normal people by using the multi-dimensional scale. We examined the differences between paranoid ideation and guarded ideation, because these ideations represented delusional ideation. The multi-dimensional scale has seven dimensions: resistance, unpleasant feeling, conviction, incorrigibility, preoccupation, dismissibility and reassurance. This scale was administered to 471 college students. The results demonstrated higher scores on resistance, unpleasant feeling, preoccupation, dismissibility and reassurance than that of guarded ideation. On the other hands, guarded ideation showed higher score on conviction and incorrigibility. These findings seemed similar to previous studies on patients with delusions. The results of present study may be indicative of the continuity view of psychosis. PMID- 15112512 TI - [Within-session changes in responding to water reinforcement: effects of variability in the duration of reinforcement]. AB - This study tested the hypothesis (McSweeney & Swindell, 1999) that habituation contributes to within-session decreases in drinking. Six rats' lever-pressing was reinforced by water under a continuous reinforcement (CRF) schedule. During the fixed amount sessions, duration of reinforcement was fixed at 3 seconds. Reinforcement duration varied from 1 to 5 seconds, with a mean of 3 seconds, during variable amount sessions. Experimental phase lasted 10 successive days and consisted of 5 fixed amount and 5 variable amount sessions, alternating day by day. Within-session decreases in responding were steeper during fixed amount than variable amount sessions. In addition, response rates were well described as linear functions of cumulative number of reinforcements. The regression line for the fixed amount sessions had steeper slope and smaller x-axis intercept than those for the variable amount sessions. These results support the habituation hypothesis and are not explained by post-ingestive factors. PMID- 15112513 TI - Response to Beggs' viewpoint. PMID- 15112514 TI - Minimally invasive dentistry: the new standard of care. PMID- 15112515 TI - Army revamps soldiers' rations in effort to promote oral health. PMID- 15112516 TI - States finding ways to make Medicaid thrive. PMID- 15112517 TI - The manual toothbrush and oral hygiene. PMID- 15112518 TI - Managing regulated waste in dental environments. PMID- 15112519 TI - When perception equals reality. What is white, what is light, what is natural? PMID- 15112520 TI - Predictable contacts with composite resins. PMID- 15112521 TI - Criteria for the replacement of defective restorations. PMID- 15112522 TI - Dental digital photography in '04. PMID- 15112523 TI - Access to success, Part 2. Improving your quality, speed, and efficiency. PMID- 15112524 TI - Endodontic hybridization. Access and answers. PMID- 15112525 TI - Immediate, direct rehabilitation of fractured maxillary central incisors in the pediatric patient. PMID- 15112526 TI - Diagnosis and treatment planning for implant placement. PMID- 15112527 TI - Considerations for establishing and maintaining proper occlusion in the aesthetic zone. PMID- 15112528 TI - Lasers in dentistry. An overview. PMID- 15112529 TI - Exophytic gingival lesions. Review of clinical and histologic features, and 3 case reports. PMID- 15112530 TI - A 12-year-old's guide to treatment acceptance. PMID- 15112531 TI - Operatory computers. Expense or investment? PMID- 15112532 TI - "Is" and "ought": moral judgments about the world as understood. PMID- 15112533 TI - From mechanical to autonomous agency: the relationship between children's moral judgments and their developing theories of mind. PMID- 15112534 TI - The role of mental state understanding in the development of moral cognition and moral action. PMID- 15112535 TI - Altruism, prudence, and theory of mind in preschoolers. PMID- 15112536 TI - Bridging the gap between theory of mind and moral reasoning. PMID- 15112537 TI - Mind and morality. PMID- 15112538 TI - Supported membrane nanodevices. AB - Supported membrane nanodevices are based on natural or artificial ion channels embedded in a lipid membrane deposited on a chip wafer. Membrane conductance is modulated by biorecognitive events, with the use of intrinsic binding sites of the ion channel or via artificial sites fused to the channel protein. Artificial ion gates are constructed by coupling a specific ligand for the analyte near the channel entrance or a site important to triggering channel conformation. The binding event leads to the closure of the ion channel or induces a conformational change of the channel, reducing the ion flux. The signal transduced from the device is the decrease in the ion flux-induced electron current at a silver silver chloride electrode at ultimate single-molecule sensitivity. Among the natural ion channels, gramicidin A, a transport antibiotic, was found to be most suitable, and thus was used by AMBRI, Australia, to set up prototypes of membrane biochips, using self-association of the dimer. Covalent dimerization-based devices, developed by the Vienna group, make use of the down-regulation of the permanently open membrane-spanning bisgramicidine ion channel. The reactive group at the C-terminus, a hydroxy group, allows precise coupling of the analyte binding moiety in gramicidin as well as bisgramicidin. The device is set up with bilayer membranes deposited on apertures of a hydrophobic frame structure produced via microlithography, facing an aqueous or hydro-gel micro-environment on both sides, constructing black lipid membranes or patch-clamp devices "on chip." The setup of the device needs gel membrane supports that allow membrane formation and contribute to the stability of the bilayer by exposure of functional groups that promote electrostatic interaction and formation of hydrogen bridges and enable the introduction of covalent spacers and anchors. Photo-cross-linked polyvinylpyrrolidone and polyacrylamide, electropolymerized polydiaminobenzene and coated agarose, as well as various chemical modifications of these polymers, were employed as membrane supports. With optimized assemblies, the membrane support did allow the formation of stable bilayer membranes, proved by "gigaseal" (electrical sealing with giga-ohm resistance) to be free of any point defects in the lipid assembly. Supports with and without hydrophilic and hydrophobic anchors were studied with reference to promoting the formation of a self-assembled membrane, to their electric resistance, and to the capability to insert functional ionophores. All components, including novel chemically engineered ion channels, novel amphiphilic lipids, a microlithographically designed chip, isolating polymer frames, and a hydrogel membrane support, are combined in the new bionanodevice. Sensitivity and specificity were proved, for example, with the use of an antibody-antigen couple down-regulating the ion flux through the membrane channel. Single ion channels incorporated in the supported lipid bilayer gave stable signals at an operational stability of several hours, which is already sufficient to test and screen for membrane receptors but still insufficient to use this device as a sensor for off-site application. Further optimization to increase operational and storage stability is done by a number of groups to allow a broad application of these devices. PMID- 15112539 TI - Template-assisted nano-patterning: from the submicron scale to the submolecular level. AB - Regular pattern formation based on template synthesis is overviewed from the submicron scale to the submolecular level. Size of patterns of course depends on size of template. Specificity and precision of interaction between the templates and between patterned materials are apparently correlated with the size and precision of patterns. PMID- 15112540 TI - Boron carbonitride nanotubes. AB - A comprehensive understanding of the design, synthesis, characterization, and properties of boron carbonitride nanotubes (BCN) is presented in this review. Distinctive structural and electronic properties are revealed in theoretical studies of the BCN nanotubes and compared with the properties of carbon nanotubes. In the experimental studies, BCN nanotubes have been synthesized by various techniques. For different purposes, controllable growth processes have been used to fabricate BCN nanotubes with novel structures, such as nanojunctions and filled nanotubes. Some interesting phenomena originating from the substitution of B and N atoms, such as the phase segregation, are considered theoretically and experimentally. Mainly the physical properties--field electron emission and photoluminescence--are discussed, which turn out to have potential applications in the industry. PMID- 15112541 TI - Recent advances in polymer nanofibers. AB - Polymer nanofibers, with diameters in the nanometer range, possess larger surface areas per unit mass and permit easier addition of surface functionalities compared with polymer microfibers. Hence, polymer nanofiber mats are being considered for use as filters, scaffolds for tissue engineering, protective clothing, reinforcement in composite materials and sensors. Although some of these applications are in the development stage, a few have been commercially exploited. Research on polymer nanofibers, nanofiber mats, and their applications has seen a remarkable growth over the last few years. However, a review of the various issues related to these nanofibers has not been published. This article presents a review of the recent trends in the processing methods and characterization techniques for polymer nanofibers. Research challenges and future trends in the processing and characterization of polymer nanofibers are discussed in the article. Five processing methods have been examined in this review, namely drawing, template synthesis, phase separation, self-assembly, and electrospinning. Among these methods, electrospinning has been used to convert a large variety of polymers into nanofibers and may be the only process that has the potential for mass production. The structure, morphology, and geometry of nanofibers and the porosity and tensile properties of nanofiber mats can be investigated through conventional techniques and instruments. But new techniques are needed for the mechanical testing of single nanofibers. Although measurement of mechanical properties such as tensile modulus, strength, and elongation is difficult because of the small diameters of the fibers, these properties are crucial for the proper use of nanofiber mats. PMID- 15112542 TI - Enhanced third-order optical nonlinearity of silver nanoparticles with a tunable surface plasmon resonance. AB - Poly(vinylalcohol) (PVA)-stabilized silver nanoparticles were prepared by a seeding method. Nanoparticles of varying morphology were obtained by controlled variation of the reaction conditions, and this method allowed the tailoring of the position of the surface plasmon resonance. The samples show two bands in the visible absorption spectrum: one in the 410-440-nm region and a second peak between 500 and 600 nm. This tunable surface plasmon resonance serves to increase the third-order optical nonlinearity (chi 3) of the nanoparticles (measured at 532 nm) by a factor of 16. PMID- 15112543 TI - Mobility of carbon nanotubes in high electric fields. AB - The influence of electric fields on carbon nanotubes is experimentally demonstrated. Alignment of nanotubes along field lines, directed motion of nanotubes between electrodes separated by several thousand micrometers, and impressive solid-state actuation behavior of nanotube-embedded structures are demonstrated, taking into account the polarization and charging of the nanotubes. These effects are reported for long strands of nanotubes, nanotubes dispersed on substrates, and nanotube-embedded polymer strips. The relative magnitude of the field responsible for polarization and directed motion was found to be dependent on the morphology of the nanotubes used. These observations may foreshadow novel electromechanical applications for nanotube elements. PMID- 15112544 TI - Synthesis and characterization of silica-embedded iron oxide nanoparticles for magnetic resonance imaging. AB - In this communication, a conceptually new approach to the delivery of magnetic resonance imaging (MRI) contrast agents is presented. Our experiments demonstrate the feasibility of using silica-embedded iron oxide nanoparticles as contrast agents in magnetic resonance imaging, where a reduction in signal intensity (increased contrast) in the T2-weighted images is observed. The surface of these particles can be chemically modified by attachment of polyethylene glycol molecules, which are found to reduce nonspecific protein binding. The design of the nanoparticle is universal and flexible and allows for facile addition or interchange of its active components (i.e., MRI contrast agents and targeting moiety) with photodynamic dyes. PMID- 15112545 TI - Interaction of thermally pretreated carbon nanomaterials with water vapor. AB - We used temperature-programmed desorption-mass spectrometry to verify whether low molecular-weight products can form by reacting thermally pretreated single-walled and multi-walled carbon nanotubes (SWNTs and MWNTs, respectively) with water vapor. The reactivity of MWNTs toward water is similar to the reactivity of graphite, whereas acid-oxidized SWNTs behave like polymerized C60 fullerene. We think the main factors influencing the reactivity are molecular surface curvature and the presence of pyrolyzable defect groups, which create highly strained bonds upon their elimination. PMID- 15112546 TI - Modification of SiO2 nanowires with metallic nanocrystals from supercritical CO2. AB - Through hydrogen reduction of metal precursors in supercritical CO2, Cu, and Pd, nanocrystals were deposited onto SiO2 nanowires to form different types of nanostructured materials, including nanocrystal-nanowire, spherical aggregation nanowire, shell-nanowire composites, and "mesoporous" metals supported by the framework of nanowires. This supercritical fluid deposition technique is an attractive approach for modifying nanowires because of its generality and simplicity; the modified nanowires could be useful as catalysts and for further fabrication of multifunctional composites. PMID- 15112547 TI - Original magnetic alignment of a nematic phase containing single-walled nanotubes. AB - Dispersions of single-walled carbon nanotubes in a 4-pentyl-4'-cyano-biphenyl nematic liquid crystal matrix are studied by X-ray scattering. For some mixtures, the addition of nanotubes strongly modifies the properties of the matrix: under a magnetic field, the nematic director is aligned perpendicular to the field, whereas it is parallel to it for pure 4-pentyl-4'-cyano-biphenyl. This original modification of liquid crystal alignment by the addition of nanotubes raises new questions about nanotubes and liquid crystal interactions. PMID- 15112548 TI - Wave packet transmission in a ZnO nanorod under the influence of repulsive/attractive scattering center. AB - We study electron wave packet propagation and scattering in ZnO nanorods. By solving the time-dependent three-dimensional Schrodinger equation, we were able to describe propagation and dynamics of the scattering process of the wave packet by ionized impurities. Scattering behavior of the wave packet by an attractive/repulsive scattering center is clearly demonstrated. PMID- 15112549 TI - Preparation of nanocrystalline alpha-Al2O3 using plant fiber. AB - Nanocrystalline alpha-Al2O3 ceramic powders have been prepared from aqueous poly vinyl alcohol (PVA)-aluminum nitrate solution and jute (plant fiber). Soluble Al ion-PVA solution formed a uniform coating on the surface of jute once it dried completely. Slow hydrolysis of aluminum ion with ammonium hydroxide deposited aluminum hydroxide on the jute surface. Decomposition of the dried aluminum hydroxide-coated jute at high temperature (1150 degrees C/2 hrs) resulted in the formation of single-phase, nanocrystalline alpha-Al2O3 with the corresponding average X-ray and TEM particle size approximately 50 nm. Precursor and heat treated powders have been characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) analysis. This method can be used to produce high-temperature as well as low-temperature nanocrystalline oxides. PMID- 15112550 TI - Tuning the setup of sputter-coated multilayers in nanocluster-based signal enhancing biochips for optimal performance in protein and DNA assays. AB - In biochip development two issues are critical: stable and specific immobilization of the ligand and achievement of high signal-to-background ratio. In this work we have addressed these issues for the development of biochips, produced by sputtering multilayers of thin metal films, metal oxides, and metal nitrides (tens to hundreds of nanometers thick) onto glass wafers. Optimized surfaces have shown good results in genomic and proteomic experiments with biochips based on surface-enhanced fluorescence and absorption techniques. PMID- 15112551 TI - Novel transducers for nano-optical biosensor chips based on biological and synthetic polymers with analyte-dependent swelling/shrinking behavior. AB - Analyte-dependent swelling/shrinking properties of ultrathin polymer layers are an appropriate means for the detection of various analytes. Optical metal nanoclusters can be used to determine the change of the layer's thickness, which is shown by a change in the color of the chip. By using different cross-linking agents and different polymers (biological or artificial as well) it was possible to design various sensitive layers showing different swelling/shrinking behaviors. Sensitivity on various analytes could be observed, since the different types of polymers employed differed in structure, functional groups, or biorecognitive properties. PMID- 15112552 TI - Formation of nanoparticle arrays on S-layer protein lattices. AB - Crystalline bacterial cell surface layers (S-layers) composed of identical protein units have been used as binding templates for well-organized arrangements of nanoparticles. Isolated S-layer proteins were recrystallized into monomolecular arrays on solid substrates (such as silicon wafers and SiO2-coated grids) and in suspension forming so-called self-assembly products. These S-layer assemblies were studied by atomic force microscopy and transmission electron microscopy (TEM). The orientation of the S-layer lattice, exhibiting anisotropic surface properties, on the solid surface and on the self-assembly products, was compared with the orientation on the bacterial cell. On both bacterial cells and SiO2 surfaces the outer face of the S-layer protein was exposed. On the self assembly products occasionally the inner face was also visible. Metal- and semiconductor nanoparticles 2 to 10 nm in mean diameter were covalently or electrostatically bound to the solid-supported S-layers and self-assembly products. TEM studies reveal that upon activation of carboxyl groups in the S layer lattice with 1-ethyl-3,3'(dimethylaminopropyl)carbodiimide (EDC), a close packed monolayer of 4-nm amino-functionalized CdSe nanoparticles could be covalently established on the S-layer lattice. Because of electrostatic interactions, anionic citrate-stabilized Au nanoparticles (5 nm in diameter) formed a superlattice at those sites where the inner face of the S-layer lattice was exposed. In contrast, cationic semiconductor nanoparticles (such as amino functionalized CdSe particles) formed arrays on the outer face of the solid supported S-layer lattices. PMID- 15112553 TI - Optical visualization of polymer-polymer interactions. AB - Surface-enhanced absorption (SEA) has been used to monitor interactions of biomolecules and to observe structural changes of polymers. Such interactions of biomolecules are of interest in high-throughput screening and diagnostics. The structural changes of synthetic polymers can be exploited in optical sensorics. Based on SEA we have developed a technique that enables us to transfer the interaction of polyelectrolytes between two flat, solid surfaces into a visible color change, exploiting nanoscopic metal clusters. The polyelectrolytes are coated onto electrochemically oxidized aluminum, glass, and polycarbonate substrates. The change of surface color is monitored with a flat-bed scanner, and the interactions are analyzed kinetically. The results are interpreted with regard to adhesion, surface tension, electrostatic interactions, and miscibility. PMID- 15112554 TI - Mesoporous SnO2 synthesized with non-ionic surfactants as an anode material for lithium batteries. AB - Mesoporous tin oxides were successfully prepared with the use of non-ionic surfactants, Pluronic 123 (P-123) and Tetronic 908 (T-908). Surface analysis of SnO2 synthesized with P123 showed an average pore size of approximately 15 nm. The high-resolution transmission electron microscopy and low-angle X-ray diffraction (XRD) measurements also confirmed their mesoporous nature. The X-ray photoelectron spectroscopy and wide-angle XRD studies revealed that these compounds have a casserite-type SnO2 structure. The electrochemical properties of these materials as anodes in lithium batteries showed excellent performance with good reversibility. The first-cycle reversible capacity was 1026 mAh/g for the material synthesized with P123. There was a high first-cycle irreversible capacity for SnO2 synthesized with the use of both non-ionic surfactants. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were employed to study the electrode kinetics during the lithium insertion process in the first cycle. There was a decrease in the charge-transfer resistance with respect to the discharge potential. The synthesis and structural and electrochemical properties of the mesoporous tin oxides are correlated and discussed in detail. PMID- 15112555 TI - A possible route to large-scale production of SWNTs through a combination of the substrate and floating catalyst methods. AB - We describe a possible route for large-scale synthesis of single wall carbon nanotubes (SWNTs) by a combination of the substrate and floating methods. The template prohibits metal particle aggregation, resulting in high-purity SWNT growth, and the three-dimensional floating seeded state of the template induces improved yields of SWNTs in a semi-continuous system. The SWNTs obtained by this method exhibit large variations of texture (isolated tubes and bundles) and a wide range of diameters (0.4-4 nm). PMID- 15112556 TI - A solvothermal route to ZnO and Mn-doped ZnO nanoparticles using the cupferron complex as the precursor. AB - ZnO nanoparticles have been synthesized from the cupferron complex by a solvothermal route in toluene solution. The nanoparticles have been prepared in the presence of various capping agents, of which the best results were obtained with tri-n-octylphosphine oxide, polyethylene glycol, and sodium bis (2 ethylhexyl) sulphosuccinate. The particles obtained with these capping agents have diameters in the 8-14 nm range. The nanoparticles have been characterized by electron microscopy, UV absorption spectroscopy, and photoluminescence spectroscopy, besides x-ray diffraction. Optical spectra of the small nanoparticles show evidence for quantum confinement. ZnO nanoparticles doped with 5% and 10% Mn could be prepared by the solvothermal route starting with a Zn(1 x)Mnx cupferron complex, and the Mn-doped nanoparticles remain paramagnetic down to 5K. PMID- 15112557 TI - Room temperature hydrogen gas sensitivity of nanocrystalline pure tin oxide. AB - Nanocrystalline (6-8 nm) tin oxide (SnO2) thin film (100-150 nm) sensor is synthesized via sol-gel dip-coating process. The thin film is characterized using focused ion-beam microscopy (FIB) and high-resolution transmission electron microscopy (HRTEM) techniques to determine the film thickness and the nanocrystallite size. The utilization of nanocrystalline pure-SnO2 thin film to sense a typical reducing gas such as hydrogen, at room temperature, is demonstrated in this investigation. The grain growth behavior of nanocrystalline pure-SnO2 is analyzed, which shows very low activation energy (9 kJ/mol) for the grain growth within the nanocrystallite size range of 3-20 nm. This low activation energy value is correlated, via excess oxygen-ion vacancy concentration, with the room temperature hydrogen gas sensitivity of the nanocrystalline pure-SnO2 thin film sensor. PMID- 15112558 TI - Zinc sulfide nanocoating of silica submicron spheres using a single-source method. AB - A chemical method to coat submicron silica particles with nanocrystalline zinc sulfide is described. The ZnS nanocoating was obtained by the thermalysis of zinc diethyldithiocarbamate in the presence of morphologically well defined silica particles. The powders obtained were isolated and were characterized by UV/visible optical reflectance, scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD), and X-ray photoelectron spectroscopy (XPS). The results showed that the silica surfaces became homogeneously coated with ZnS as the reaction time was increased. This coating consists in clusters of small ZnS particles whose dimensions are within the nanometric range. PMID- 15112559 TI - Bucky-wires and the instability of diamond (111) surfaces in one-dimension. AB - Recent advances in the fabrication and characterization of semiconductor and metallic nanowires are meeting the high expectations of nanotechnolgists. Although diamond has remarkable electronic and chemical properties, development of diamond nanowires has been slow, while the development of carbon nanotube based technologies continues at a furious pace. Recently, the theoretical and experimental observation of the transformation of nanodiamonds into carbon-onions (and vice versa) has led to a new intermediate phase of carbon, denoted "bucky diamond", with a diamond core encased in an carbon onion-like shell. These findings lead to the question of whether a similar transformation occurs in diamond nanowires. We used ab initio techniques to determine the relaxed structure of diamond nanowires with octahedral surface facets, with results exhibiting delamination of octahedral surfaces, and indicating the formation of "bucky-wires". The effects of surface hydrogenation upon this transition also is examined. PMID- 15112560 TI - Synthesis of germanium oxide nanoparticles in low-pressure premixed flames. AB - Germanium oxide (GeOx) nanoparticles in the size range from 1 nm to 5 nm were synthesized in a low-pressure premixed H2/O2/Ar flat flame of 30 mbar. The premixed flame was doped with different amounts of tetramethyl germanium (Ge(CH3)4) ranging from 250 ppm to 2000 ppm. The influence of process parameters such as the amount of oxygen in the reaction gas, the condensation and reaction time, standoff, and precursor concentration with respect to growth of germanium oxide particles were investigated. The particles formed were analyzed in situ according to their mass and charge with a particle mass spectrometer. The specific surface area was determined ex situ by the BET method. The crystal form and chemical composition of produced nanopowders were characterized by EDX analysis and X-ray diffraction measurements. PMID- 15112561 TI - Characterization of nanoparticles of LiMn2O4 synthesized by a one-step intermediate-temperature solid-state reaction. AB - Nanoparticles of lithium manganese oxide (LiMn2O4) with a spinel structure have been synthesized by a one-step intermediate temperature solid-state reaction. The influence of the molar ratio of citric acid to the metal ions on the physicochemical properties of LiMn2O4 powders in air has been analyzed by means of X-ray diffraction and electron microscope techniques. The electrochemical behavior of the material has been examined by charge/discharge tests and cyclic voltammetry. Test results reveal that LiMn2O4 particles with lower molar ratios of citric acid to metal ions (1:2) are highly crystalline and highly electrochemically reversible, with better cycle capabilities when compared with a sample with a higher molar ratio (2:1). The LiMn2O4 powders obtained by this method have a uniform morphology with a narrow size distribution. PMID- 15112562 TI - The phenomenon of changing coiling-chirality in carbon nanocoils obtained by catalytic pyrolysis of acetylene with various catalysts. AB - Carbon nanocoils were prepared by the chemical vapor deposition process of the catalytic pyrolysis of acetylene at 700-800 degrees C with various catalysts. Twisting or coiling-formed carbon nanocoils with changing coiling-chirality and zigzag-formed carbon nanofibers were obtained with SUS 304, WS2, Pt-Pd, TiN, and Ni as the catalysts. Their morphologies and microstructures were examined in detail, and then the changing mechanism of the coiling-chirality was discussed. No apparent difference in the microstructure between the part of a nanocoil with changing coiling-chirality and the part of a zigzag nanofiber with changing zigzag-chirality, or between a bulk right-clockwise coil and a bulk left clockwise coil was observed. It was supposed that changing coiling-chirality was mainly caused by the gradual or successive change in chemical composition on the thin layers present on the surface of catalyst grains during the chemical vapor deposition process. PMID- 15112563 TI - Ultrasound-assisted fabrication of nanoporous CdS films. AB - A new method for fabricating nanoporous CdS films is reported. It involves exposing the CdS solution with ultrasound waves during the process of dip coating. Indium tin oxide (ITO)-coated glass and plastic (commercial transparency) were used as substrates. In each case three different precursors were used for dip coating. The precursors used were CdCl2 and thiourea in one case and CdS nanoparticles prepared by sonochemical and microwave-assisted methods in the other two cases. X-ray diffraction studies performed on these powders show a phase corresponding to cubic CdS. The Field Emission Scanning Electron Microscopy (FE-SEM) images of the films on plastic showed uniform pores with a diameter of 80 nm for all three methods. Optical absorption measurements indicated a blue shift and multiple peaks in the absorption curve. The FE-SEM observations of the films on an ITO/glass substrate indicated a crystalline film with voids. The UV-vis absorption results indicated a blue shift in the absorption with an absorption edge at 435, 380, and 365 nm for CdS films made by solution growth, sonochemical, and microwave routes, respectively. The magnitude of the absorption is dependent on film thickness, and the observed blue shift in the absorption can be explained on the basis of quantum confinement effects. PMID- 15112564 TI - Catalytic growth of carbon nanoballs with Co encapsulation from CH4 decomposition: MoOx-promoted shrinking of the carbon nanoball size. AB - Compared with Co/MgO catalysts with high Co loading and without Mo addition, the addition of Mo oxide to low Co-loading Co/MgO catalysts can promote the growth of smaller sized Co-encapsulated carbon nanoballs (CNBs) by the decomposition of methane. The catalysts were carefully studied by X-ray photoelectronspectroscopy (XPS), X-ray diffraction (XRD), and temperature-programmed reduction (TPR). The promotion effect of Mo on the increase in the carbon deposition and the reduction in the CNB size, as well as the relation between the carbon product and metal particle size, are discussed. PMID- 15112565 TI - Formation of pile networks by long carbon nanotubes from decomposition of CO on Co-Mo film. AB - We report the formation of pile networks by long carbon nanotubes grown at 700 degrees C from a Co-Mo film on a quartz plate. Carbon monoxide (CO) was used as the carbon source. The networks were formed because the density of catalyst particles on the substrate was low, which resulted in low carbon nanotube density that did not support vertical growth. At the same time, the low carbon nanotube density makes it possible for CO to reach the catalysts on the substrate for continuous growth. No obvious amorphous carbon chunks were observed, suggesting that the pile networks consisted of fairly high-quality, long carbon nanotubes. PMID- 15112566 TI - Nanocomposite magneto-rheological fluids with uniformly dispersed Fe nanoparticles. AB - A systematic study of the magnetic and rheological properties of magneto rheological (MR) fluids containing micron-size and nano-size iron particles is presented. The MR fluids were prepared with hydraulic oil as the carrier liquid and lecithin as an effective surfactant medium that promotes uniform particle dispersion. Magnetic measurements on micron-, hybrid-(nano + micron), and nano-MR fluids clearly indicate that the partial replacement of the micro-size particles by nanoparticles results in a better suspension and robust chain formation under applied external magnetic fields. For nano-MR fluids, the measured yield stress was found to be lower than micron-MR fluids. However, better flow properties and sharper magnetic switching make nanoparticle-based MR fluids appealing for microfluidics device applications where higher yield stress is not required. PMID- 15112567 TI - Microemulsion processing of silica-polymer nanocomposites. AB - Silica-polymer nanocomposites have been synthesized via polymerizable bicontinuous microemulsions to disperse functionalized/nonfunctionalized SiO2 nanoparticles uniformly in a polymerized microemulsion system as revealed by transmission electron microscopy. The effect of both types of SiO2 nanoparticles in the polymerized microemulsion increased its glass transition temperature (Tg) from 90 to 128 degrees C. TgS of functionalized silica nanocomposites were about 15 degrees C higher than those of nonfunctionalized samples. As expected, both hardness and modulus of the nanocomposites increased with the loading of both types of silica from 2 to 6 wt%. In the case of nonfunctionalized silica, the improvement of properties may be simply exerted by the conventional filler effect due to the better dispersion of nanoparticles of silica in fluid microemulsion prior to the polymerization. The additional effect for the functionalized silica containing a terminal vinyl group is that It could be grafted to the polymer matrix through cross-polymerization with microemulsion-formed polymer to establish strong nanocomposite networks. PMID- 15112568 TI - Precipitation phase transformation in nanocrystalline Fe-Mo alloys. AB - Precipitation phase transformation was studied in nanocrystalline Fe-rich Fe-Mo alloys with the use of X-ray diffraction and Mossbauer spectroscopy. Alloys up to 5 at% Mo in Fe were synthesized by mechanical alloying and formed in alpha phase bcc solid solutions with average grain sizes in the range of 10-13 nm. The precipitation transformation (alpha-->alpha + lambda) was found to proceed via a Mo clustering that was correlated with the size of the nanograins. This was understood in terms of the Gibbs Thomson effect with a concept of negative surface energy contribution to the Gibbs free energy of mixing in a nanocrystalline alloy with positive internal energy of mixing. This contribution increased the stability of the solid solution for nanosized grains, and the Mo precipitation started once the grains grew beyond a critical size. We argue that the Mo precipitation takes place in the grain boundary regions, and the Mo-rich lambda phase also precipitates directly in the grain boundary regions, in contrast to the microcrystalline alloys, where the Mo clusters formed within the grains and were first dissolved in the Fe matrix before the lambda phase was formed. PMID- 15112569 TI - Synthesis of aligned zinc sulfide nanostructures and the influence of experimental conditions on their morphologies and phase. AB - A novel aligned flower-like array and single-crystal nanosheets composed of ZnS quantum wires were fabricated via a simple solution route. In the method, ZnCl2 chemicals reacted with Na2S in 50 ml of ethylenediamine (en) solution containing different amounts of hydrochloric acid (HCl) at 70-80 degrees C. After annealing at 500 degrees C for 1.5 h, wurtzite phase ZnS nanoscaled materials were synthesized. When the amount of HCl was 2 ml and 3 ml, respectively, flower-like structure and nanosheets were obtained. The products were characterized by X-ray diffraction, transmission electron microscopy, high-resolution electron microscopy (HREM), and electron diffraction (ED). The influences of concentration, temperature, and reaction time on the morphologies and phase of ZnS nanostructures were also studied. The photoluminescence peaks are located at approximately 308 nm and approximately 410 nm. The formation mechanism is also discussed here. PMID- 15112570 TI - Lanthanopolyoxotungstoborates: synthesis, characterization, and layer-by-layer assembly of europium photoluminescent nanostructured films. AB - New lanthanopolyoxotungstoborates, K6-xHx[Ln(BW11O39)(H2O)3].nH2O, Ln(III) = Sm, Eu, Tb, Er, were prepared and characterized by spectroscopic methods (Fourier transform infrared, Fourier transform-Raman, 11B solid-state nuclear magnetic resonance, and photoluminescence) and elemental analysis. A layer-by-layer assembly method was employed to fabricate multilayered films containing the europium heteropolyanion and the polyelectrolytes poly(sodium 4-styrenesulfonate) and poly(diallydimethylammonium chloride). The photoluminescence behavior of these final nanostructures was investigated and compared with that of the starting Eu(III) polyoxometalate used as the building unit. PMID- 15112571 TI - "Experimental oncology": achievements and prospects for the future. PMID- 15112572 TI - Poly-drug cancer therapy based on ceramide. AB - Thousands of research studies have reported that many kinds of cancer cells and tumors can be killed by treatments that increase the concentration of a simple cellular sphingolipid, ceramide (Cer). While there are many ways to elevate tumor Cer levels, this approach is complicated by the central, complex role of Cer in cell homeostasis: Cer is readily metabolized to form other sphingolipids that increase the tumor's growth rate, metastasis, and resistance to the patient's immune system. This review points out the need to prevent this metabolic conversion while simultaneously stimulating the enzymes that increase the formation of Cer. I describe here many of the enzymes that need stimulation or inhibition, and drugs or metabolites or dietary components that modify each of the enzymes. The review also points to the importance of the allylic alcohol group in Cer and in many cancer drugs, suggesting that the hydroxyl group participates in phosphate transfer to and from proteins by forming a temporary phosphate ester. The allylic hydroxyl may also reduce the ketone moieties in mitochondrial ubiquinone, with formation of reactive oxygen species and apoptogenic breakdown. The level of Cer in tumors can be increased by: (1) direct administration of Cer or a Cer analogue, and (2) stimulation of Cer synthesis from its elementary precursors, or from (3) sphingomyelin by hydrolysis, or from (4) the glucosphingolipids by hydrolysis, or (5) by acylation of sphingosine. In addition, Cer concentration can be raised by slowing its conversion to (6) sphingomyelin, (7) glucosylCer, (8) Cer phosphate, and (9) sphingosine + fatty acid by hydrolysis. Therapeutic radiation stimulates the de novo synthesis of Cer in tumors. Conversion of sphingosine (from Cer) to sphingosine phosphate probably also ought to be blocked. PMID- 15112573 TI - Venous thromboembolism in oncology. AB - Thromboembolic events represent well-recognised complications of neoplastic disease contributing, in a significant manner, to the morbidity and mortality from cancer. The close relationship between the activation of blood coagulation and tumor growth is known since 1865, when Armand Trousseau first described the clinical association between primary or idiopathic venous thromboembolism and an underlying occult malignancy. However, only in the last decades significant advances in this field have been achieved, both on the comprehension of the complex interactions between the tumor and the hemostatic system, and on the prophylaxis and therapy of the thromboembolic manifestations in cancer patients. PMID- 15112574 TI - Identification of a novel binding partners for tumor suppressor PTEN by a yeast two-hybrid approach. AB - AIM: To identify novel PTEN-binding partners. METHODS: The technique of yeast two hybrid screening was used in this study. A panel of bait constructs was created, containing the C-terminal domain of PTEN, full length PTEN, activated and phosphatase-dead mutants. The expression of LexA-fused baits, their nuclear localization and autoactivation potential were tested according to the standard protocol of Duplex A system. CDNA libraries from Colon Cancer, HeLa and Mouse Embryo were screened with two selected bait constructs. Isolated positive clones were further analysed by mating assay and identified by automated DNA sequencing and database searching. RESULTS: Extensive screening of cDNA libraries with the full length and the C-terminal domain of PTEN led to the identification of 43 positive clones, which were confirmed in mating assay. Sequence analysis indicated that two clones encode AEBP1 (Adipocyte Enhancer Binding Protein 1). CONCLUSION: Our data indicate that the interaction between PTEN and AEBP1 is mediated by their C-terminal and N-terminal domains, respectively. The functional importance of PTEN-AEBP1 interaction is currently under investigation. PMID- 15112575 TI - Effect of 2'-deoxyisoguanosine on the growth of human tumor and normal cell lines. AB - AIM: To test the effect of 2'-deoxyisoguanosine as a specific substrate of HIV reverse transcriptase on the growth of cultured normal and cancer cells. METHODS: The effect of 2'-deoxyisoguanosine on the growth of cells of two normal (telomerase-negative) and five cancer cell lines have been tested. Cell viability was assessed by MTT assay. RESULTS: We have found that this nucleoside analogue has low potency and specificity in inhibiting tumor cell growth. IC50 ranged from 0.5 mM to 2 mM for tumor cells, and from 1 mM to more than 4 mM for normal cells. CONCLUSIONS: 2'-deoxyisoguanosine has low potency and specificity in inhibiting tumor cell growth, similar to other telomerase inhibitors. PMID- 15112576 TI - Immunohistochemical analysis of S6K1 and S6K2 expression in human breast tumors. AB - AIM: To express recombinant S6K2 in baculovirus expression system; to purify large quantities of recombinant S6K2 for biochemical studies; to generate and characterise specific MABs against recombinant S6K2; to study the patterns S6K1 and S6K2 expression and subcellular localization in normal, benign and malignant breast tissues. METHODS: Recombinant baculovirus, expressing wild type S6K2 was generated using Bac-to-Bac system (Invitrogen); recombinant S6K was purified from infected Sf9 cells using affinity purification approach; monoclonal antibodies against recombinant S6K2 were generated; the specificity of generated MABs towards recombinant and endogenous S6K2 were examined by ELISA, Western blotting, immunoprecipitation and immuhohistochemical staining; immunohistochemical detection of S6K1 and S6K2 in human breast tissues was performed using specific monoclonal antibodies towards S6K1 and S6K2. RESULTS: Large amounts of enzymatically active S6K2 were purified using baculovirus expression system; highly purified preparations of S6K2 were used to generate and characterize anti S6K2 MABs; elevated levels of S6K1 and S6K2 were found in breast tumors when compared to normal breast tissues; S6K2 is frequently localized in the nuclei of adenocarcinoma tissues, but rarely in fibroadenoma or "normal" breast tissues. CONCLUSION: Production of recombinant S6K2 in large amount and generation of specific monoclonal antibodies towards S6K2 has provided us with excellent tools to study the function and regulation of this important signalling molecule in normal and cancer cells. Immunnohistochemical analysis of S6K1 and S6K2 expression in normal and malignant breast clearly indicates that both kinases are overexpressed in breast tumors, when compared to "normal" tissues. The retention of S6K2 in the nuclei of malignant cells may be caused by disregulation of nucleocytoplasmic shuttling and could subsequently affect cell growth and proliferation. PMID- 15112577 TI - Downregulation of estrogen receptor alpha and beta expression in carcinogen induced mammary gland tumors of rats. AB - AIM AND METHODS: The recent discovery of a new isoform of estrogen receptor (ER), ER beta, has promoted the investigation of its expression on mammary gland. This study was carried out to examine the expression of ER alpha, ER beta and proliferating cell nuclear antigen (PCNA) in the carcinogen-induced mammary tumors induced by N-methyl-N-nitrosourea (MNU) or 7,12-dimethylbenz[a]anthracene (DMBA), and to compare these expression with those of age-matched normal mammary glands. RESULTS: There was significant decrease of expression of ER alpha and ER beta in the mammary gland tumors compared with age-matched normal mammary glands (p < 0.05). In mammary gland tumors, ER alpha expression was mainly located in epithelial cells, showing intranuclear staining pattern. The decrease of ER beta expression was so distant that some tumor cells did not show any expression. There was a complete loss of ER beta expression in 50% (7/14) of MNU-induced mammary gland tumors, and 68.2% (15/22) of DMBA-induced mammary gland tumors. However, there was no difference in PCNA expression between mammary gland tumors and normal mammary glands. CONCLUSION: This study represents that the decrease of expression of ER alpha and ER beta is associated with mammary carcinogenesis, and suggests that modulation of ER alpha and ER beta may be the target for the treatment of mammary gland tumors. PMID- 15112578 TI - Oxidative metabolism of rat blood in the course of alveolar hepatic carcinoma PC 1 growth. AB - AIM: To evaluate oxidative metabolism of rat blood in the course of alveolar hepatic cancer growth in vivo. METHODS: The oxidation imbalance was assessed by the rise in the values of the integral index of oxidation stress. The structural and functional state of erythrocyte membranes was investigated by spin electron paramagnetic resonance spectroscopy. RESULTS: The growth of alveolar carcinoma was found to be associated with intensification of lipid peroxidation processes with increased blood content of conjugated dienes, malonic dialdehyde against the background of decreased concentration of endogenous antioxidants tocopherol and retinol. Destabilization of the structural state of erythrocyte membranes of rat tumor hosts at the development of oxidation stress was studied, which was characterized by nonspecific structural changes of membrane sorption centres, reduction in specific capacity in the protein-lipid contact area and its increase in the phospholipid bilayer, rise in the degree of order and polarity. CONCLUSION: Alveolar carcinoma growth in rats resulted in intensification of free radical lipid peroxidation processes with a shift of the prooxidant-antioxidant balance to the left and development of oxidation stress. PMID- 15112579 TI - Nuclear factor-kappa B regulates cyclooxygenase-2 expression and cell proliferation in human colorectal carcinoma tissue. AB - OBJECTIVE: Activation of transcription factor nuclear factor-kappa B (NF-kappa B) has been shown to play a role in cell proliferation, apoptosis, cytokine production, and oncogenesis. The purpose of this study was to determine whether NF-kappa B is constitutively activated in human colorectal tumor tissues and, if so, to determine the role of NF-kappa B in colorectal tumorigenesis, furthermore, to determine the association of RelA expression with the expression of cyclooxygenase-2 (COX-2) and tumor cell proliferation. METHODS: Paraffin sections of the normal epithelial, adenomatous and adenocarcinoma tissue were analysed immunohistochemically for RelA, COX-2, Ki-67 protein expression. EMSA (electrophoretic mobility shift assay) was used to confirm the increased nuclear translocation of RelA in colorectal tumor tissues. The expression of COX-2 mRNA was determined by RT-PCR (reverse transcription polymerase chain reaction) analysis. RESULTS: Activation of NF-kappa B was significantly higher in adenocarcinoma tissue in comparison to that in adenomatous and normal epithelial tissue. The colon tumor cell proliferation, mRNA expression and protein level of COX-2 were significantly increased in the transition from normal to tumor tissue. CONCLUSION: Our results suggest that NF-kappa B may promote proliferation via enhancing the expression of COX-2, and the increased expression of RelA/nuclear factor-kappa B plays an important role in the pathogenesis of colorectal carcinoma. PMID- 15112580 TI - Characterization of a new type HPV16 E7 variant isolated from cervical cancer highest incidence area in Hubei Province of China. AB - AIM: To investigate the variation and biological properties of HPV16 E7 isolated from cervical cancer biopsy samples from highest incidence area in HuBei province of China. METHODS: HVP16 E7 sequences isolated from the cervical cancer biopsies of 10 local patients were amplified, sequenced and compared with prototype E7 gene. Then the variant gene was cloned into different vectors to study the antigenicity, expression and immunogenicity of its protein by Western blot, immunofluorescence and genetic immunization in vitro or in vivo. RESULTS: The results showed that 7 of 10 samples had the same mutations which led to a nonsense mutation at codon 43 of E7 sequence. The truncated E7 protein could be recognized by standard E7 monoclonal antibody in Western blot and expressed in NIH3T3 cells. In the blood sera of mice immunized intramuscularly by the plasmid DNA expressing the variant E7 gene specific E7 antibodies could be detected at week 2, 3, 5 and 6 after inoculation. However, no specific lymphoproliferation after E7 protein stimulation in vitro was detected by MTT colorimetric assay in comparison to the prototype E7 protein. CONCLUSION: HPV16 E7 gene may show variation in China and the variant protein could be expressed and induce host humoral immune response, but could not elicit special cellular-immune response against it. These data might hold the key for future development of HPV16 vaccine in HuBei province of China. PMID- 15112581 TI - Different antitumor immunity roles of cytokine activated T lymphocytes from naive murine splenocytes and from dendritic cells-based vaccine primed splenocytes: implications for adoptive immunotherapy. AB - AIM: The aim of the study is to explore the antitumor capacity of effector cells generated from murine splenocytes with sequential addition of a cocktail of cytokines and the possible contribution of dendritic cells to the antitumor capacity of these effector cells. METHODS AND RESULTS: Interferon-gamma, interleukin (IL)-1 beta, anti-CD3 mAb and IL-2 were used to activate murine splenocytes either from naive mice (termed cytokine activated T cells, CAT) or from DC based vaccine primed mice (termed specific effector T cells, SET). The antitumor roles of SET and CAT were analyzed in murine L615 T lymphocytic leukemia. Both CAT and SET were CD4(+)-predominant phenotypically and didn't show any significant cytotoxicity against a variety of syngeneic and allogeneic target cell lines using 51Cr release assay. When injected in vivo in combination with CY, CAT can cure a large proportion of leukemia mice. The cured mice couldn't establish specific antitumor immunity. However, in contrast to the roles of CAT, SET show far superior antitumor efficacy on a per cell basis compared with CAT. Moreover, the SET cured mice developed tumor specific long term memory immunity which was sufficient to reject a subsequent otherwise lethal tumor cells rechallenge and was transferable to naive immunocompetent mice. CONCLUSION: Our data demonstrate that there remain fundamentally different antitumor functions of CAT and SET which might be useful in the immunotherapy strategy choices. PMID- 15112582 TI - Experimental immunotherapy of mice with transplanted MC-rhabdomyosarcoma resistant to doxorubicin. AB - AIM: To compare the sensitivity of doxorubicin (DOX) sensitive and DOX-resistant MC-rhabdomyosarcoma (MC-RMS) cells to the action of lymphokine-activated cells (LAC). RESULTS: In vitro investigations showed that LAC received from the fraction of adherent lymphocytes possess the highest activity against DOX resistant tumor cells, and LAC from lymphocytes of total pool--against DOX resistant tumor cells pretreated with DOX at a low dose. Adoptive immunotherapy of MC-RMS in vivo showed the highest efficacy in the cases of LAC intratumoral injection and the one combined with intraperitoneal administration of DOX at a low dose (increase of survival time by 14% and 25%, respectively). CONCLUSION: Adoptive in vivo therapy of DOX-resistant Mh-RMS is effective if LAC or their combination with DOX at a low dose are administered. PMID- 15112583 TI - Enhancement of antitumor response to sarcoma 45 in rats by combination of whole body hyperthermia and interleukin-2. AB - AIM: To evaluate the summarized effect of hyperthermia and interleukin-2 (IL-2) administration on antitumor defense in tumor-bearing rats. METHODS: Nonbred rats after subcutaneous inoculation of sarcoma 45 cells were treated with whole-body hyperthermia (WBH, +42.5 degrees C, 60 min) and interleukin-2 (IL-2, 10,000 U/kg of body weight). Parameters of tumor growth and survival of animals were monitored till day 33 after tumor cell inoculation. RESULTS: Combined application of WBH and IL-2 at 5th day after tumor cell transplantation resulted in a delay of tumor growth and improvement of survival parameters in comparison with control group or animals that received separate treatment. Therapeutic efficacy of WBH combined with IL-2 was analogous to a single-dose chemotherapy with cyclophosphamide. CONCLUSION: Combined application of WBH and IL-2 is the useful approach for cancer immunotherapy. PMID- 15112584 TI - Effects of two different high doses of irradiation on antioxidant system in the liver of guinea pigs. AB - AIM: To examine the state of the oxidant-antioxidant system in the liver of guinea pig caused by high doses of ionizing radiation in the early period. METHODS: The research was carried out on guinea pigs irradiated with the doses of 8 Gy (group 2) or 15 Gy (group 3) (single dose/whole body) in comparison with control group (group 1). The levels of thiobarbituric acid reactive substances (TBARS) and glutathione (GSH), the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) and the levels of selenium in the liver were measured. RESULTS: TBARS levels in the irradiated animals were markedly higher than those in controls. In group 3, GSH levels and GSH-Px activity were significantly increased while activity of SOD and CAT were significantly decreased compared to groups 1 and 2. Liver selenium levels were not influenced by irradiation. CONCLUSION: The data have shown that gamma irradiation at the doses of 8 Gy or 15 Gy results in significant increase in free radical formation while antioxidant enzymes were affected only at a dose of 15 Gy. PMID- 15112585 TI - Down regulation of DRET gene in human hepatocellular carcinoma tissues samples from Qidong area, China. AB - OBJECTIVE: To analyze the expression of DRET gene in hepatocellular carcinomas (HCCs) tissue specimens in comparison with normal liver tissue to evaluate the relationship between DRET gene and HCC. METHODS: 250 primary HCCs and 50 normal liver tissue samples from Qidong area, China, were studied for DRET mRNA and protein expression with the use of Northern blot analysis, in situ hybridization and immunohistochemistry. RESULTS: By Northern analysis, moderate to strong DRET mRNA expression was present in normal liver samples. In contrast, DRET mRNA expression in tissue samples of primary HCCs was markedly decreased compared with normal controls. Primary HCCs that gave rise to metastasis showed significantly lower DRET mRNA levels than nonmetastasizing HCCs. By in situ hybridization analysis, nonmetastatic HCCs samples didn't differ from controls. In contrast, most of the primary metastasizing HCC showed only faint or moderate DRET mRNA expression. Tissue sections of nonmetastatic HCC exhibited lower DRET immunoreactivity than control samples, but higher labeling index than metastatic HCC samples. CONCLUSIONS: Expression of DRET on mRNA and protein levels in HCC cells is related to HCC metastatic ability. PMID- 15112586 TI - Detection of polyomavirus DNA in human brain tumors. AB - AIM: To investigate surgically resected human brain tumors for the presence of polyomavirus DNA. MATERIALS AND METHODS: 65 samples of surgically resected human brain tumors (57 tumors were of neuroectodermal genesis, 4--mesenchimal, 4- metastatic tumors) were examined for the presence of viral DNA by PCR analysis. RESULTS: DNA of polyomaviruses was found in oligodendroligomas (66.7% of cases), oligodendroastrocytomas (66.7% of cases) and glioblastomas (40% of cases). In metastatic tumors viral infection had not been detected. No correlation between the grade of brain tumors and infection with polyomavirus DNA has been revealed. CONCLUSION: Our data point to importance of investigation of the brain tumors and cerebrospinal fluid for determination of viral infection. PMID- 15112587 TI - Drive time. Interview by Nick Edwards. PMID- 15112588 TI - Clinical management. Where medicine meets management. Uneasy riders. AB - The NHS needs to form new alliances to address the public health challenges posed by the second Wanless report and the government's consultation paper on public health. Public health physicians need to return to clinical practice if they are to play a credible leadership role. 'Policy partnerships' could promote issues such as health inequalities. PMID- 15112590 TI - Finance. Pay and dismay. PMID- 15112589 TI - Change management. Pass it on. PMID- 15112591 TI - HSJ people. Show the way. PMID- 15112592 TI - Context-free forensic science. PMID- 15112593 TI - A critical evaluation of two methods of signature analysis. AB - The assessment of signature disguise, where an individual attempts to disguise their own signature on a document with the intent of later disclaiming it (so called 'view to deny' signatures), is a problem faced by many document examiners. This study evaluates a method known as the angle value test and another experimental method involving angle measurements to determine if either of them can reliably establish whether a questioned signature is disguised or has been written by another person. By using 29 sets of normal and disguised signatures, both methods of analysis were shown to be unreliable techniques for identifying the author of a particular signature. PMID- 15112594 TI - Absence of salting out effects in forensic blood alcohol determination at various concentrations of sodium fluoride using semi-automated headspace gas chromatography. AB - Blood alcohol measurements determined by headspace gas chromatography have been challenged on the grounds that the presence of the preservative sodium fluoride in blood samples artificially increases headspace alcohol concentrations due to a salting out effect. Blood samples containing varying amounts of ethanol and sodium fluoride were tested using semi-automated headspace gas chromatography with n-propyl alcohol as the internal standard to assess the validity of this challenge. We find, in fact, that under these test conditions the measured alcohol levels are systematically depressed as the amount of sodium fluoride in the blood sample increases. The challenge thus has no basis. PMID- 15112595 TI - Post-conviction DNA testing: the UK's first 'exoneration' case? AB - The routine incorporation of forensic DNA profiling into the criminal justice systems of the United Kingdom has been widely promoted as a device for improving the quality of investigative and prosecutorial processes. From its first uses in the 1980s, in cases of serious crime, to the now daily collection, analysis and comparison of genetic samples in the National DNA Database, DNA profiling has become a standard instrument of policing and a powerful evidential resource for prosecutors. However, the use of post-conviction DNA testing has, until recently, been uncommon in the United Kingdom. This paper explores the first case, in England, of the contribution of DNA profiling to a successful appeal against conviction by an imprisoned offender. Analysis of the details of this case is used to emphasise the ways in which novel forms of scientific evidence remain subject to traditional and heterogeneous tests of relevance and credibility. PMID- 15112596 TI - The population of coloured fibres in human head hair. AB - In 2002 a population study of textile fibres in human hair was carried out using 26 volunteers in Cambridgeshire, UK. Over 12,000 fibres were recovered from a variety of hair lengths using low adhesive tape and classified according to colour, generic type and fibre length. The results of the study showed that the most common fibre colours were black/grey (48%), blue (29.1%) and red (12.7%), the least common being green, orange/brown and yellow which each accounted for less than 5% of the total. Natural fibres (mainly cotton) were predominant (72.3%) and man-made fibres were considerably less frequent. When colour and generic type were classified together, the most common combinations were black and blue cottons. The least common were the man-made fibre/colour combinations with the most frequent of these accounting for less than 7% of the sample. Fibre loads carried by long hair were found to be significantly less than that carried by short hair. The results of this study are in accordance with previous fibre population studies using other types of recipient surfaces and are likely to be influenced by factors such as seasonal and geographical variation. PMID- 15112597 TI - Sex determination by PCR analysis of DNA extracted from incinerated, deciduous teeth. AB - Establishing the biological sex of human remains is a very important part of identifying victims of fire when severe soft tissue destruction has occurred. Deciduous (children's) teeth were exposed to a range of incineration temperatures 100-500 degrees C for 15 minutes. Polymerase Chain Reaction (PCR) amplification was used to identify specific human amelogenin regions. There was successful identification of human biological sex, from deciduous teeth exposed to incineration temperatures of 200 degrees C and below, using standard ethidium bromide gel staining. There was greater sensitivity using fragment analysis by laser induced fluorescence which achieved sex identification from some teeth heated to 400 degrees C. PMID- 15112598 TI - Forensic hair morphology comparison--a dying art or junk science? AB - There has been debate in both the judicial and forensic fields concerning the admissibility and reliability of the so-called forensic comparison sciences such as handwriting, tool mark analyses, and hair analysis. In particular, there has been increasing controversy over the use and interpretation of hair comparison evidence and it has been held partly responsible for miscarriages of justice. There has also been a perceived devaluation of the worth of microscopic human hair analysis particularly since the advent of DNA profiling. This article will attempt to initiate discussion on the past, current and future role of forensic human hair analysis and comparison. PMID- 15112599 TI - One register--one standard partnership across the forensic spectrum. PMID- 15112603 TI - Usage limiting presents huge challenges. PMID- 15112604 TI - Top level system developed. AB - Much attention in the healthcare engineering sector is focused on how the NHS programme for improving decontamination services will roll out. Shaping up is the way in which the private sector will be involved and in this article Nicholas Marshall examines a company's advanced system for cleaning and sterilising surgical instruments. PMID- 15112606 TI - Ventilation in healthcare premises. PMID- 15112605 TI - Off-site construction: a key prescription. PMID- 15112607 TI - Aiming to liberalize provision of services. PMID- 15112609 TI - New concept introduced. PMID- 15112608 TI - Cost-effectively automating fire door release. AB - UK care home fire statistics are the worst for 40 years. After catastrophic fires in Scotland and Wales this year the death toll in care homes rose to 18 fatalities over a single month. No larger loss of life in a fire at a care home in Britain has been recorded since regulations covering these homes were introduced in the 1960s. It is against this background of heightened national vigilance that Fireco's Chris Pearce explains, in a timely overview, how the very latest "add-on" automatic fire door release technology can revolutionize integrated fire alarm systems by easing access in the care setting. PMID- 15112610 TI - Avoiding workplace violence incidents. PMID- 15112611 TI - Adding a creative edge. PMID- 15112612 TI - Improvements in ionisation. PMID- 15112613 TI - New priorities and challenges for epidemiology in the Americas. PMID- 15112614 TI - Global alert: severe acute respiratory syndrome (SARS). PMID- 15112615 TI - Country profiles. Bolivia. PMID- 15112616 TI - Country profiles. Guyana. PMID- 15112617 TI - Country profiles. Haiti. PMID- 15112618 TI - Country profiles. Honduras. PMID- 15112619 TI - Country profiles. Nicaragua. PMID- 15112620 TI - Absence of transmission of the D9 measles virus in the region of the Americas, November 2002 - May 2003. PMID- 15112622 TI - Case definitions. Ebola-Marburg viral diseases. PMID- 15112621 TI - Techniques to measure the impact of mortality: years of potential life lost. PMID- 15112623 TI - Case definitions. Onchocerciasis (river blindness). PMID- 15112624 TI - Diagnosis on the use of bulletins for the dissemination of epidemiological information in the region of the Americas. PMID- 15112625 TI - A time capsule of health situation analysis in the Americas, 1902-2002. PMID- 15112626 TI - Severe acute respiratory syndrome (SARS) update. PMID- 15112627 TI - Organization and status of civil registration and vital statistics in countries of the English-speaking Caribbean. PMID- 15112628 TI - Recommendation for the monitoring of measles eradication in the American region. PMID- 15112630 TI - Case definitions. American trypanosomiasis (Chagas' disease). PMID- 15112631 TI - Case definitions. African trypanosomiasis (sleeping sickness). PMID- 15112632 TI - On the estimation of mortality rates for countries of the Americas. PMID- 15112633 TI - Life tables: a technique to summarize mortality and survival. PMID- 15112634 TI - Revision of the International Health Regulations. PMID- 15112635 TI - [Salary raises of dentists and dental technicians in South Africa]. PMID- 15112636 TI - [In memory of Jean-Pierre Soulier, the writer]. PMID- 15112637 TI - [Liberalization of legal advertising: legal internet access by the physician]. PMID- 15112639 TI - Bibliography. Current world literature. Seizure disorders. PMID- 15112638 TI - Acute metabolic cataract as a first manifestation of diabetes mellitus in a 12 year-old girl. PMID- 15112640 TI - Bibliography. Current world literature. Metabolic disorders and neurotoxicology. PMID- 15112641 TI - Fusion inhibitor trials halted. PMID- 15112642 TI - Boosted saquinavir approved. PMID- 15112643 TI - Infection rates increase for Native Americans. PMID- 15112644 TI - Drug users embrace Vancouver site. PMID- 15112645 TI - Human subjects protections, institutional review boards, and cultural anthropological research. PMID- 15112646 TI - Haemophilus influenzae type b meningitis in children, Eritrea. PMID- 15112647 TI - Quinolone safety and efficacy more important than potency. PMID- 15112649 TI - Vancouver facing syphilis outbreak. PMID- 15112648 TI - Vancomycin-resistant Enterococcus faecalis in Serbia. PMID- 15112650 TI - Automatic system approved. PMID- 15112651 TI - Heterosexuals still not being tested. PMID- 15112652 TI - Q fever in Como, Northern Italy. PMID- 15112653 TI - The RGS (regulator of G-protein signalling) and GoLoco domains of RGS14 co operate to regulate Gi-mediated signalling. AB - RGS (regulator of G-protein signalling) proteins stimulate the intrinsic GTPase activity of the a subunits of heterotrimeric G-proteins, and thereby negatively regulate G-protein-coupled receptor signalling. RGS14 has been shown previously to stimulate the GTPase activities of Ga(o) and Ga(i) subunits through its N terminal RGS domain, and to down-modulate signalling from receptors coupled to G(i). It also contains a central domain that binds active Rap proteins, as well as a C-terminal GoLoco/G-protein regulatory motif that has been shown to act in vitro as a GDP-dissociation inhibitor for Ga(i). In order to elucidate the respective contributions of the three functional domains of RGS14 to its ability to regulate G(i) signalling, we generated RGS14 mutants invalidated in each of its domains, as well as truncated molecules, and assessed their effects on G(i) signalling via the bg pathway in a stable cell line ectopically expressing the G(i)-coupled M2 muscarinic acetylcholine receptor (HEK-m2). We show that the RGS and GoLoco domains of RGS14 are independently able to inhibit signalling downstream of G(i). Targeting of the isolated GoLoco domain to membranes, by myristoylation/palmitoylation or Rap binding, enhances its inhibitory activity on G(i) signalling. Finally, in the context of the full RGS14 molecule, the RGS and GoLoco domains co-operate to confer maximal activity on RGS14. We therefore propose that RGS14 combines the inhibition of G(i) activation or coupling to receptors via its GoLoco domain with stimulation of the GTPase activity of Ga(i) GTP via its RGS domain to negatively regulate signalling downstream of G(i). PMID- 15112654 TI - [Extrafascial excision of the rectum or total meso-rectal excision?]. AB - Total mesorectal excision is a valuable technique in preventing local recurrences cancer. However the use of the word "mesorectum" is inaccurate anatomically, and the implication that total excision of all the perirectal fat contained within the perirectal fascia in all patients with rectal cancer will minimize local recurrences remains contentious. The term extrafascial excision of the rectum is more accurate. He may contribute as well to a better understanding of the surgical technique allowing all surgeons to improve their own results. PMID- 15112655 TI - Natural product glycorandomization. AB - glycorandomization is a chemoenzymatic strategy that overcomes the limitations in natural product derivatization associated with both solely chemistry-based approaches or in vivo engineering. In this article we present the basic strategies for glycorandomization development as a next-generation tool in drug discovery. PMID- 15112656 TI - [Should early feeding be permitted following elective colorectal resection?]. PMID- 15112657 TI - [Patient information. Prophylaxis of hepatitis A]. PMID- 15112658 TI - Guideline-based care cuts mortality in elderly heart attack patients. PMID- 15112659 TI - Studies evaluate hysterectomy as treatment for abnormal uterine bleeding. PMID- 15112660 TI - Preparation period for post-thyroid cancer surgery test can be shorter, study says. PMID- 15112661 TI - Long-term use of Fosamax sustains or continues increases in bone density. PMID- 15112662 TI - [Endoprostheses of the knee joint]. AB - Due to the advances in prosthesis design and standardisation of implant techniques, the maintenance of implants in arthroplasty of the knee joint has enormously increased during the past few years. Various arthroplastic designs enable an adjustment towards the patient's individual indiction and anatomy. For one-compartmental arthrosis, monocondylous prosthesis can be implanted using microinvasive operation techniques. Prerequisite for mere joint resurfacing is an intact lateral ligament apparatus, whereas the the symmetrical ligament balancing is essential for long-term analgesia, joint stability and good flexion. Aseptic loosening, discreet infections, instabilities and patellar problems are common reasons for painful knee arthroplastics. PMID- 15112663 TI - [Treatment of oral dryness in Sjogren's syndrome]. AB - PURPOSE: In Sjogren's syndrome (SS), oral dryness is frequently the most bothersome symptom of sicca syndrome with negative affects on quality of life. A review of treatments of oral dryness is proposed. CURRENT KNOWLEDGE AND KEY POINTS: To date, so specific DMARD has demonstrated its efficacy in SS. Hydroxychloroquine is frequently used but did not demonstrate any clinical benefit in te only small randomized control study versus placebo available. Thus, the only treatments are symptomatic. The most recent data show that systemic cholinergic agonist (pilocarpine and cevimeline) are effective in the symptomatic treatment of dryness. Pilocarpine (Salagen) is the only systemic cholinergic agonist available in Europe. It has been agreed in France since July 2003. FUTURE PROSPECTS AND PROJECTS: Use of immunosuppressive drugs may be useful in some complications of SS. Unfortunately, promising results from an open study with infliximab (Remicade) were not confirmed by a large randomized control study involving more than 100 patients. New control studies with old drugs such as hydroxychloroquine, or new ones such as rituximab, are mandatory. PMID- 15112664 TI - [Legal aspects of medical further education]. PMID- 15112665 TI - User-controlled mapping of significant literatures. AB - We apply a version of our web-based literature-mapping system to PNAS for 1971 2002, as indexed by the National Library of Medicine and the Institute for Scientific Information. Given a single input term from a user, a medical subject heading, a cocited author, or a cocited journal, PNASLINK rapidly displays views in which that term and the other 24 terms that most frequently co-occur with it in a bibliographic database are interrelated in ways suggesting fruitful combinations for document retrieval. The interrelationships are produced by two algorithms, pathfinder networks and Kohonen-style self-organizing maps. PNASLINK displays are themselves interactive interfaces that can retrieve documents from digital libraries (e.g., PNAS Online). This style of visualizing knowledge domains is called "localized" because it does not attempt to map the indexing of literatures in full but concentrates on the top terms in an "associative thesaurus" reflecting user interests. It also permits swift remappings, as the user recognizes terms worth pursuing. PNASLINK is illustrated with maps drawn from the literature of population genetics. Some comparative and evaluative comments are added, one from a domain expert indicating that the face validity of the system may be tempered by insufficient specificity in the indexing terms being mapped. PMID- 15112666 TI - Bacillus anthracis incident, Kameido, Tokyo, 1993. PMID- 15112667 TI - Virus taxonomy: one step forward, two steps back. PMID- 15112668 TI - Phenotypic variability: clinical presentation between the 6th year and the 60th year in a family with X-linked agammaglobulinemia. PMID- 15112669 TI - Occupational asthma and IgE sensitization to 7-aminocephalosporanic acid. PMID- 15112670 TI - Potential adverse effects of the inhaled corticosteroids. PMID- 15112671 TI - Abstracts of the Annual Main Meeting of the Society for Experimental Biology. 29 March-2 April 2004, Edinburgh, United Kingdom. PMID- 15112672 TI - The "gianic faces" of nitric oxide in asthma: role for the inducible and the constitutive nitric oxide synthase isoforms. PMID- 15112673 TI - Misuse of terminology to imply that 1,25-dihydroxy-vitamin D is a nutrient: there is no evidence for an association between vitamin D and allergy. PMID- 15112674 TI - Abstracts of the 45th Annual Meeting of the Japanese Society of Plant Physiologists. Tokyo, Japan, March 27-29, 2004. PMID- 15112675 TI - New Trends in Diabetes Management. Abstracts of the 10th Annual Conference of the UDOP, EHF, PAHO/WHO, and CFNI. March 4-7, 2004, Ocho Rios, Jamaica. PMID- 15112676 TI - Abstracts of the 4th Hellenic Forum on Bioactive Peptides. April 22-24, 2004, Patras. PMID- 15112677 TI - Coronary heart disease--ACE inhibitors for all patients? PMID- 15112679 TI - Abstracts of the International Conference on Applied Genomics, 9th ESACP/16th ISDQP Meeting. Amsterdam, The Netherlands, 1-4 October 2003. PMID- 15112678 TI - The almost forgotten biological potentials of cholesterol and its intermediates: cholesterol reduction and myositis. PMID- 15112680 TI - Crosslinked polyethylene standard aids effort to improve surgical implants. PMID- 15112681 TI - High-risk insurance pools: safety net or tightrope? PMID- 15112682 TI - Children's behavioral health screening. PMID- 15112683 TI - Controlled-release oxycodone for the management of pediatric postoperative pain. AB - Studies addressing pain management after pediatric spinal fusion surgery have focused on the use of patient-controlled or epidural analgesia during the immediate postoperative period. Controlled-release (CR) analgesics have been found to be safe and effective in adults. The purpose of this study was to describe the use of oxycodone-CR in pediatric patients after the immediate postoperative period. A retrospective chart review of 62 postoperative spinal fusion patients (10-19 years) was conducted. The mean initial oxycodone-CR dose was 1.24 mg/kg/day. The mean ratio of conversion from parenteral morphine equivalents to oxycodone-CR was 1:1. Mean pain scores decreased from 4.2/10 to 3.7/10 with the transition to oxycodone-CR. Common side effects included dizziness, constipation, and nausea. Oxycodone-CR was used for an average of 13.3 days, which included an average wean time of 6 days. Results of this study demonstrate safe and effective use of oxycodone-CR in the pediatric spinal fusion population. PMID- 15112684 TI - Human subjects protection and cultural anthropology. PMID- 15112685 TI - Trials and tribulations of navigating IRBs: anthropological and biomedical perspectives of "risk" in conducting human subjects research. PMID- 15112686 TI - Assisted suicide. PMID- 15112687 TI - Jewish law perspectives on suicide and physician-assisted dying. PMID- 15112688 TI - The obligation to heal and patient autonomy in Jewish law. PMID- 15112689 TI - Jewish perspectives on assisted suicide and euthanasia. PMID- 15112690 TI - Democrat presidential hopefuls unveil plans for US health care. Candidates outline proposals to improve access to health care--one of the top issues for US voters. PMID- 15112691 TI - Suicide, assisted suicide and euthanasia: a Buddhist perspective. PMID- 15112692 TI - A response to Damien Keown's Suicide, assisted suicide and euthanasia: a Buddhist perspective. PMID- 15112693 TI - A response to Damien Keown's Suicide, assisted suicide and euthanasia: a Buddhist perspective. PMID- 15112694 TI - Health research and human rights in South Africa. PMID- 15112695 TI - HIV treatment in South Africa: overcoming impediments to get started. PMID- 15112696 TI - Medical education after the first decade of democracy in South Africa. PMID- 15112697 TI - The autonomy imperative of behavioral research. PMID- 15112698 TI - Immigration consequences of tuberculosis. PMID- 15112699 TI - The impotency of the potentiality argument for fetal rights: reply to Wilkins. PMID- 15112700 TI - Why potentiality cannot matter. PMID- 15112701 TI - China's shift in HIV/AIDS policy marks turnaround on health. Pledge to provide free HIV tests shows China is starting to take action on combating the disease. PMID- 15112703 TI - Abstracts of the Aerospace Medical Association 75th Annual Scientific Meeting. May 2-6, 2004, Anchorage, Alaska, USA. PMID- 15112702 TI - Abstracts of the X National Congress of Digestive Diseases, Italian Federation of Digestive Diseases. Turin, Italy, 27-31 March 2004. PMID- 15112704 TI - Abstracts of the German Society of Cell Biology Annual Meeting. Berlin, Germany, 24-27 March 2004. PMID- 15112705 TI - Indian experts protest against off-label drug use. PMID- 15112706 TI - Defining the microsatellite instability phenotype in colorectal cancer through analysis of surrogate markers. PMID- 15112707 TI - Ghana battles drug-resistant malaria with artesunate. PMID- 15112708 TI - WHO waters down draft strategy on diet and health. PMID- 15112709 TI - [Abstracts of the 4th International Conference on Hypertension and Kidney. Madrid, Spain, 12-14 February 2004]. PMID- 15112710 TI - An interest in human dignity as the basis for genomic torts. PMID- 15112711 TI - Misuse of genetic information: the common law and professionals' liability. PMID- 15112712 TI - Abstracts of the 92nd Annual Meeting of the Poultry Science Association. July 6 9, 2003, Madison, Wisconsin, USA. PMID- 15112713 TI - Pharmacologic and clinical characteristics of thrombolytic agents. AB - Arterial and venous thromboembolic events, including myocardial infarction, ischemic stroke, peripheral arterial thrombosis, deep venous thrombosis, and pulmonary embolism are common potentially life-, organ-, and limb-threatening vascular diseases. Anticoagulant therapy is recommended in these settings to prevent further thrombosis pending gradual clearance of the thrombotic occlusion by the endogenous fibrinolytic system. Recognition of the importance of the fibrinolytic system in thrombus resolution has resulted in the development of pharmacologic fibrinolytic (thrombolytic) agents to facilitate rapid restoration of vascular patency. Several plasminogen activator (PA) thrombolytic agents with different pharmacokinetic and pharmacodynamic properties have been developed to treat thrombotic disease. Newer PAs have been developed as "fibrin-specific", bolus-administration drugs to primarily treat acute coronary syndromes. Continuous infusions of these fibrin-specific PAs have become popular for the lysis relatively larger peripheral vascular thromboses. Loss infusion of newer tissue-type plasminogen activator-based PAs may result in an increased risk of bleeding, including intracranial hemorrhage. Currently available data fail to provide compelling evidence that newer PAs offer significantly greater efficacy and safety than well-established agents like urokinase when used to treat peripheral vascular thrombosis. PMID- 15112714 TI - [Olanzapine in therapy of a somatoform disorder]. PMID- 15112715 TI - Families: the forgotten resource for individuals with mental illness. PMID- 15112716 TI - Adjustment difficulties of adolescents with sickle cell disease. AB - TOPIC: The psychosocialfactors that affect adolescents with sickle cell disease (SCD). PURPOSE: To explore whether specific psychosocial factors can provide clues to the future adjustment of this population. SOURCES: Ovid Web, Medline, Psychinfo, and CINAHL databases for the years 1997 to 2001. CONCLUSIONS: Promoting effective psychosocial functioning is as important as managing the medical aspects of SCD, yet this is an area of care that is commonly overlooked. Nurses, therefore, have an opportunity to have a significant impact on the lives of adolescents with SCD if they intervene in ways to promote both biological and psychosocial adjustment. PMID- 15112717 TI - Using poetry to discover and share significant meanings in child and adolescent mental health nursing. AB - TOPIC: Does writing and reading poetry help nurse-therapists, students, and clients reflect on significant meanings and gain a greater understanding of intense clinical situations? PURPOSE: To describe the use of poetry in managing intense feelings, discuss the relevant literature, articulate how students and clients responded to the use of poetry, and address the limitations of such an approach. SOURCES: Published literature, clinical expertise, and poems written by the author. CONCLUSIONS: Writing and reading poetry helps clinicians, students, and clients give voice to situations that touch their hearts. Poetry and literature should be used more extensively in clinical and educational settings. PMID- 15112718 TI - Identifying competence in the context of the pediatric anogenital exam. AB - PROBLEM: Appraisals of and concerns about the pediatric anogenital exam in order to promote a better understanding of predisposing factors that may underlie some people's adjustment to these exceptional circumstances. METHODS: Thirty-three girls ages 8-15 and 82 adult guardians rated their acute exam appraisals and concerns using a 5-point Likert-type scale. Data were collected immediately after the anogenital exam at a pediatric outpatient clinic specializing in medical evaluations following allegations of child sexual abuse. FINDINGS: Participants reported relatively low levels of acute exam concerns as well as relatively high appraisals of the exam's importance and desirability, their ability to emotionally tolerate the exam, and optimism regarding the event's potential amelioration. CONCLUSIONS: Data suggested the presence of specific competencies that may be instrumental in safeguarding children's and adult guardians' psychological well-being in the context of the pediatric anogenital exam. PMID- 15112719 TI - Relationships among personal space boundaries, peer acceptance, and peer reputation in adolescents. AB - PROBLEM: Peer relationships are important to the health of adolescents who are at a developmental period of moving from parental influence to potentially dangerous peer influence. Interpersonal communication processes are logical antecedents of peer relationships. However, research in this area with adolescent populations is limited. METHODS: Exploratory descriptive study using observational technique to collect nonverbal and verbal communication data on adolescents (N = 28). Observer ratings of personal boundaries and peer acceptance were correlated with anonymous peer ratings of sociability/leadership, aggressiveness/disruptiveness, and sensitivity/isolation, with comparisons made between males and females. FINDINGS: Boundaries are related to peer acceptance and peer reputation and are interpreted differently in males and females. CONCLUSIONS: This study provides beginning evidence of the usefulness of Personal Space Boundary Theory in understanding adolescent behavior. PMID- 15112720 TI - Where is the "community" in community treatment? PMID- 15112721 TI - Nurses lead in linkages for health and juvenile justice systems. PMID- 15112722 TI - Acquired and partially de novo synthesized pyrrolizidine alkaloids in two polyphagous arctiids and the alkaloid profiles of their larval food-plants. AB - The profiles of pyrrolizidine alkaloids (PAs) in the two highly polyphagous arctiids Estigmene acrea and Grammia geneura and their potential PA sources in southeastern Arizona were compiled. One of four species of Boraginaceae, Plagiobothrys arizonicus, contained PAs; this is the first PA record for this plant species. The principle PA sources are Senecio longilobus (Asteraceae) and Crotalaria pumila (Fabaceae). The known PA pattern of S. longilobus was extended; the species was found to contain six closely related PAs of the senecionine type. Three novel PAs of the monocrotaline type, named pumilines A-C, were isolated and characterized from C. pumila, a species not studied before. The pumilines are the major PAs in the seeds, while in the vegetative organs they are accompanied by the simple necine derivatives supinidine and as the dominant compound subulacine (1beta,2beta-epoxytrachelanthamidine). In both plant species, the PAs are stored as N-oxides, except C. pumila seeds, which accumulate the free bases. Great variation in PA composition was observed between local populations of C. pumila. The PA profiles were established for larvae and adults of E. acrea that as larvae had fed on an artificial diet supplemented with crotalaria-powder and of G. geneura fed with S. longilobus. In both experiments, the larvae had a free choice between the respective PA source and diet or food plants free of PAs. The profiles compiled for the two species reflect the alkaloid profiles of their PA sources with one exception, subulacine could never be detected in E. acrea. Besides acquired PAs, insect PAs synthesized from acquired necine bases and necic acids of insect origin were detected in the two arctiid species. These insect PAs that do not occur in the larval food sources accounted for some 40-70% (E. acrea) and 17-37% (G. geneura) of total PAs extracted from the insects. A number of novel insect PAs were identified. Plant-acquired and insect PAs were found to accumulate as N-oxides. The results are discussed in relation to specific biochemical, electrophysiological, and behavioral mechanisms involved in PA sequestration by arctiids. PMID- 15112723 TI - The role of methyl salicylate in prey searching behavior of the predatory mite phytoseiulus persimilis. AB - Many carnivorous arthropods use herbivore-induced plant volatiles to locate their prey. These plant volatiles are blends of up to hundreds of compounds. It is often unknown which compounds in such a complex volatile blend represent the signal to the foraging carnivore. We studied the role of methyl salicylate (MeSA) as part of the volatile blend in the foraging behavior of the predatory mite Phytoseiulus persimilis by using a Y-tube olfactometer. MeSA is one of the compounds released by lima bean, infested with Tetranychus urticae--a prey species of the predatory mite. MeSA attracted satiated predatory mites in a dose dependent way with optimum attraction at a dose of 0.2 microg. Predatory mites did not discriminate between a prey-induced lima bean volatile blend (that contains MeSA) and a prey-induced volatile blend to which an extra amount of synthetic MeSA had been added. However, they preferred a MeSA-containing volatile blend (induced by T. urticae) to an otherwise similar but MeSA-free blend (induced by jasmonic acid). Adding synthetic MeSA to the MeSA-free blend significantly increased the mites' choice for this odor, suggesting an important role for MeSA. This study is a new step toward unraveling the role of herbivore induced plant volatiles in the foraging behavior of predatory arthropods. PMID- 15112724 TI - The role of honeydew in host searching of aphid hyperparasitoids. AB - Foraging in many insect parasitoids is mediated by chemicals associated with hosts. For example, honeydew, the feces of feeding aphids, induces and/or prolongs searching behavior of aphid parasitoids. In the laboratory, we tested if aphid hyperparasitoids, which belong to a higher trophic level, also rely on aphid honeydew to locate their hosts. We used the potato aphid. Macrosiphum euphorbiae, the primary parasitoid, Aphidius nigripes, and four hyperparasitoids, Asaphes suspensus, Dendrocerus carpenteri. Alloxysta victrix, and Syrphophagus aphidivorus that possess different biological attributes and host ranges. We determined if foraging hyperparasitoid females could discriminate between (i) honeydew from a host and a non-aphid host (the potato aphid and the soft brown scale, Coccus hesperidum), and (ii) honeydew from healthy aphids and those parasitized by A. nigripes. Females of A. suspensus did not react to any of the honeydew treatments. While the presence of non-aphid honeydew did not modify the behavior of A. victrix, D. carpenteri, and S. aphidivorus females, they exhibited an increase in searching time and path length but not walking speed when in the presence of honeydew from aphids. However, there were no changes in host searching behaviors, such as antennation or ovipositor probing that have been reported for primary aphid parasitoids. There was no difference in the response of hyperparasitoid females to honeydew from healthy and parasitized aphids. These results indicate that hyperparasitoids may use aphid honeydew, a conspicuous cue from the second trophic level, as an infochemical to locate their hosts. PMID- 15112725 TI - Hydroxybenzoic acid derivatives in a nonhost rutaceous plant, Orixajaponica, deter both oviposition and larval feeding in a rutaceae-feeding swallowtail butterfly, Papilio xuthus L. AB - A Rutaceae-feeding swallowtail butterfly. Papilio xuthus L., feeds on various rutaceous plants but always rejects Orixa japonica Thunb. (Rutaceae). Females were strongly deterred from laying eggs by a methanolic extract of O. japonica leaves. Larvae also rejected a diet leaf medium impregnated with O. japonica leaf extracts. Several components in the water-soluble fraction of the leaf extract were found to deter both oviposition and feeding responses. Two major deterrent compounds were characterized as 5-[[2-O-(beta-D-apiofuranosyl)-beta-D glucopyranosyl]oxy]-2-hydroxybenzoic acid and adisyringoyl aldaric acid. These compounds induced potent deterrence of both oviposition and larval feeding by P. xuthus, which suggests a congruent chemosensory mechanism of allomonal chemicals acting on both female tarsal chemoreceptors and larval maxillary taste receptors. PMID- 15112726 TI - Rapid herbivore-induced changes in mountain birch phenolics and nutritive compounds and their effects on performance of the major defoliator, Epirrita autumnata. AB - Insect damage changes plant physiology and chemistry, and such changes may influence the performance of herbivores. We introduced larvae of the autumnal moth (Epirrita autumnata Borkh.) on individual branches of its main host plant. mountain birch (Betula pubescens ssp. czerepanovii (Orlova) Hamet-Ahti) to examine rapid-induced plant responses, which may affect subsequent larval development. We measured systemic responses to herbivory by analyzing chemistry, photosynthesis, and leaf growth, as well as effects on larval growth and feeding, in undamaged branches of damaged and control trees. Larvae reared on leaves from intact branches of the herbivore-damaged trees grew faster than those reared on leaves of control trees, indicating systemic-induced susceptibility. Herbivore damage did not lead to systemic changes in levels of primary nutrients or phenolic compounds. The analyses of photosynthetic activity and individual hydrolyzable tannins revealed a reversal of leaf physiology-herbivore defense patterns. On control trees, consumption by E. autumnata larvae was positively correlated with photosynthetic activity: on damaged trees, this correlation was reversed, with consumption being negatively correlated with photosynthetic activity. A similar pattern was found in the relationship between monogalloylglucose, the most abundant hydrolyzable tannin of mountain birch, and leaf consumption. Among the control trees, consumption was positively correlated with concentrations of monogalloylglucose, whereas among herbivore-damaged trees, this correlation was reversed and became negative. Our results suggest that herbivore performance is related to both concentrations of phenolic compounds and photosynthetic activity in leaves. This linkage between herbivore performance, leaf chemistry, and physiology was sensitive to induced plant responses caused by slight herbivore damage. PMID- 15112727 TI - Toxic effects of lemon peel constituents on Ceratitis capitata. AB - A series of experiments were conducted to evaluate the toxicity of lemon peel extracts incorporated into mediterranean fruit fly Ceratitis capitata diet. Extracts were obtained with different solvents: diethyl ether, ethyl acetate, and methanol. All three extracts were toxic to some extent; the diethyl ether extract was selected for further studies. Ether extracts of lemon peel were prepared weekly over a 2-month period, from fruits collected on the 1st d of the bioassay. Weekly GC-MS and UV analyses of the extracts demonstrated that the concentration of citral and coumarins decreased in the peel after harvest. We conducted a series of bioassays to evaluate the toxicity of the ether extract, and mixtures of this extract with citral, 5,7-dimethoxycoumarin. and linalool incorporated to C. capitata larvae's natural diet (lemon slices endocarp) at a concentration of 250 microg/g of diet. Significant larvicidal activity can be obtained from a fresh lemon peel extract; however, when the extract was obtained from stored lemons, toxicity decreased. Addition of small amounts of citral or 5,7 dimethoxycoumarin, and linalool to the stored lemon peel extract would bring back the toxicity to the rates of fresh lemons extracts. Finally, female adults of C. capitata fed on diets containing additional amounts of ether extract, 5,7 dimethoxycoumarin, and linalool, were exposed to different photoperiods to test for phototoxicity. The treatment was toxic and affected the oviposition capacity of females depending on photoperiod. PMID- 15112728 TI - Host-plant specialization in pheromone strains of the European corn borer Ostrinia nubilalis in France. AB - European corn borer (ECB) feeding on maize (Zea mais), mugwort (Artemisia vulgaris), and hop (Humulus lupulus) are genetically different in France and referred to as host-plant races. Here, we investigated sex pheromone composition as a possible trait linked to the host plant. ECB host races were sampled from 13 different sites in France. GC-MS analysis of female pheromone showed that 175 out of 176 maize females belonged to the Z type with one hybrid. In contrast, mugwort and hop females belonged almost exclusively to the E type. No Z females were found on these plants and only 2 females out of 169 were hybrids. In the three sites of sympatry, the hybrid proportion was far from Hardy-Weinberg expectations. Wind tunnel experiments showed that 76-79% of maize males from three populations were attracted by Z females, whereas neither mugwort nor hop males were. Mugwort males from Toussus-le-Noble were attracted by E females originating from an American maize strain. These data showed that maize, mugwort, and hop host races of O. nubilalis differ not only in their host plant but also in the sex pheromone they use. Because mugwort and hop are putative ancestral host plants, these results are discussed from the point of view of evolutionary scenarios for the emergence of Z and E strains. PMID- 15112729 TI - Attraction of the larval predator Elater ferrugineus to the sex pheromone of its prey, Osmoderma eremita, and its implication for conservation biology. AB - Elater ferrugineus is a threatened click beetle inhabiting old hollow trees. Its larvae consume larvae of other saproxylic insects including the threatened scarab beetle Osmoderma eremita. Recently, (R)-(+)-gamma-decalactone was identified as a male-produced sex pheromone of O. eremita. Here we present evidence that E. ferrugineus adults use this odor as a kairomone for location of their prey. In field trapping experiments, significantly more trapping events of E. ferrugineus beetles were observed in Lindgren funnel traps baited with (R)-(+)-gamma decalactone than in control traps (20 vs. 1, respectively). Analyses of headspace collections from E. ferrugineus beetles indicate that the predator itself does not produce the substance. Both sexes were attracted to the prey pheromone. suggesting that E. ferrugineus males use the odor as an indirect cue for location of mates or of the tree hollows, which make up their habitat. When compared to pitfall traps, the Lindgren system was significantly more effective in trapping E. ferragineus, and no difference could be established for O. eremita, showing the high potential to use odor-based systems to catch both species. We suggest that (R)-(+)-gamma-decalactone could be used as a master signal in monitoring programs for these vulnerable beetle species. which are both regarded as indicators of the associated insect fauna of the threatened habitat of old hollow trees. PMID- 15112730 TI - Cuticular hydrocarbons and aggression in the termite Macrotermes subhyalinus. AB - Cuticular hydrocarbons are among the prime candidates for nestmate recognition in social insects. We analyzed the variation of cuticular hydrocarbons in the termite species M. subhyalinus in West Africa (Comoe National Park) on a small spatial scale (< 1 km). We found considerable variation in the composition of cuticular hydrocarbons among colonies, with four distinct chemical phenotypes. Different phenotypes occurred within each of the four habitats. The difference between these phenotypes is primarily due to unsaturated compounds. A clear correlation between the difference of the hydrocarbon composition and the aggression between colonies was found. This correlation also holds in a multivariate analysis of genetic similarity (measured by AFLPs), morphometric distances (measured by Mahalanobis-distances). as well as geographic distances between colonies. In a more detailed analysis of the correlation between the composition of cuticular hydrocarbons and aggression, we found that no single compound is sufficient to explain variation in aggression between pairings of colonies. Thus, termites seem to use a bouquet of compounds. Multiple regression analysis suggested that many of these compounds are unsaturated hydrocarbons and, thus, may play a key role in colony recognition. PMID- 15112731 TI - Male aggregation pheromone of date palm fruit stalk borer Oryctes elegans. AB - Laboratory and field investigations were carried out to characterize the chemical communication system of the date palm fruit stalk borer, Oryctes elegans, and to develop pheromone-based trapping in Eastern Iran. Adults of both sexes feeding on date palm pieces attracted conspecifics, whereas date palm alone was minimally attractive. Males were twice as attractive as females. More beetles were captured at the palm crown than at ground level. Odors from adults feeding on sugarcane were sampled and analyzed by gas chromatography and mass spectrometry. Whereas females did not emit sex specific volatiles, males emitted a blend of 4 methyloctanoic acid (1: major component) and ethyl 4-methyloctanoate (2), occasionally mixed with minor components: 4-methyloctanyl acetate (3), methyl 4 methyloctanoate (4), 4-methyloctanol (5), and nonanyl acetate (6). Electroantennography and field trapping experiments demonstrated that compound 1 is an essential component of the male aggregation pheromone of O. elegans. It was barely attractive by itself but synergistic with fresh date palm odor. It attracted many more beetles than any of compounds 2-6. The addition of one or several of compounds 2-6 to 1 did not improve trap captures. During the course of 2 years, we captured 4000 beetles, with a weekly average of 6.3 beetles/trap, and were able to monitor the seasonal flight of O. elegans. Our results provide the basis for developing mass trapping for control of this pest. PMID- 15112732 TI - Analysis of anal secretions from phlaeothripine thrips. AB - The anal secretions of 16 phlaeothripine thrips species (Thysanoptera: Phlaeothripidae) were studied, including a reinvestigation of three species previously reported. A total of 37 components were detected, including hydrocarbons, acetates, terpenes, carboxylic acids, a quinone, an aromatic compound, and a pyranone compound. The secretions of all species were composed of some of these components, with Xylaplothrips inquilinus possessing as many as 11 components. Of these components, (Z)-9-octadecene, (Z)-9-nonadecene, nonadecadiene, octanoic acid, decanoic acid, geranial, neral, alpha-pinene, beta pinene, caryophyllene, 2-hydroxy-6-methylbenzaldehyde, and two unidentified monoterpenes [UK-I (M+136) and UK-II (M+168)] were detected for the first time. The chemicals were species-specific; four Liothrips species and three Holothrips species could be distinguished from each other and their congeners by the GC profiles of the ether extracts of their anal secretions. The anal secretions of gall-inducing thrips commonly contained terpenes. of which citral (a mixture of geranial and neral) and beta-acaridial repelled ants or had antifungal activity. The findings suggest that these terpenes play a defensive role and prevent galls from fungal infestation. 3-Butanoyl-4-hydroxy-6-methyl-2H-pyran-2-one, found from three Holothrips spp., caused paralysis in ants. Chemical analysis of anal secretion components is a useful method for the classification of tubuliferan species that are difficult to distinguish on the basis of morphological characters. PMID- 15112733 TI - Zapoteca formosa: sulfur chemistry and phytotoxicity. AB - Chemical analysis of Zapoteca formosa extracts show that a variety of volatile sulfur-containing components, including cyclic polysulfides, are responsible for the distinctive odor of its germinating seeds and young plants. The major sulfur containing compounds include acetyl djenkolic acid, djenkolic acid, taurine, cystine, benzothiazole, dimethyl disulfide, 2,4-dithiapentane, 2 hydroxyethylmethylsulfide, trithiane, 1,2,4-trithiolane, and 1,2,4,6 tetrathiepane. Decreased amounts of djenkolic acid in seedlings and young plants, as compared to the seed, indicate that this is likely the metabolic precursor of the volatile sulfur components. Germination and radicle elongation assays show that germinating seeds of Zapoteca and mixtures of volatile sulfur-containing compounds are highly phytotoxic to Cucumis sativa, Lactuca sativa, Lycopersicon esculentum, and Acacria farnesiana. PMID- 15112734 TI - Casuarina cunninghamiana tissue extracts stimulate the growth of Frankia and differentially alter the growth of other soil microorganisms. AB - Aqueous extracts of host plant Casuarina cunninghamiana tissue altered the in vitro growth of its diazotrophic microsymbiont Frankia and a selection of other soil microorganisms. The growth of actinomycetous Frankia strains, 55005. AvcI1, CesI5, CjI82 001, and Cj was stimulated by aqueous extracts of C. cunninghamiana tissue. Green cladodes (photosynthetic branches), unsuberized roots, and suberized roots were more stimulatory than dry cladodes and seed tissue. Aqueous extracts of green cladodes of C. cunninghamiana most stimulated the growth of Casuarina-derived Frankia strains CjI82 001 and 55005. The growth of isolates of soil bacteria Bradyrhizobium japonicum, Arthrobacter globiformis and Bacillus subtillis and of the soil fungi Penicillium oxalicum and Arthroderma cookiellum was either inhibited or not affected by cladode extracts. Cladode extracts stimulated the growth of the actinomycete Streptomyces albus and the fungus Rhizopus homothallicus. The magnitude (as great as 100%) of the increase in growth caused by tissue extracts for the Casuarina-derived Frankia strains relative to other soil microbes suggests a host-specific enhancement of the microsymbiont. PMID- 15112735 TI - Juglone disrupts root plasma membrane H+-ATPase activity and impairs water uptake, root respiration, and growth in soybean (Glycine max) and corn (Zea mays). AB - Juglone is phytotoxic, but the mechanisms of growth inhibition have not been fully explained. Previous studies have proposed that disruption of electron transport functions in mitochondria and chloroplasts contribute to observed growth reduction in species exposed to juglone. In studies reported here, corn and soybean seedlings grown in nutrient solution amended with 10, 50, or 100 microM juglone showed significant decreases in root and shoot dry weights and lengths with increasing concentrations. However, no significant differences in leaf chlorophyll fluorescence or CO2-dependent leaf oxygen evolution were observed, even in seedlings that were visibly affected. Disruption of root oxygen uptake was positively correlated with increasing concentrations of juglone, suggesting that juglone may reach mitochondria in root cells. Water uptake and acid efflux also decreased for corn and soybean seedlings treated with juglone, suggesting that juglone may affect metabolism of root cells by disrupting root plasma membrane function. Therefore, the effect of juglone on H+-ATPase activity in corn and soybean root microsomes was tested. Juglone treatments from 10 to 1000 microM significantly reduced H+-ATPase activity compared to controls. This inhibition of H+-ATPase activity and observed reduction of water uptake offers a logical explanation for previously documented phytotoxicity of juglone. Impairment of this enzyme's activity could affect plant growth in a number of ways because proton-pumping in root cells drives essential plant processes such as solute uptake and, hence, water uptake. PMID- 15112736 TI - Stereochemistry of host plant monoterpenes as mate location cues for the gall wasp Antistrophus rufus. AB - In spring, adult males of the gall wasp Antistrophus rufus L. emerge from inconspicuous galls in stems of their host plant Silphium laciniatum L. and search for sites on stems where females will later emerge. The behavior of males suggests that they use olfaction rather than visual or tactile cues in searching for mates. In an earlier publication, we reported that galls of A. rufus were associated with changes in enantiomeric ratios of alpha- and beta-pinene emitted by plant stems, and hypothesized that monoterpene stereochemistry served as a mate location cue for adult males. Here, we support this hypothesis with bioassays that demonstrate that males can discriminate between galled and ungalled stems, as well as between blends of synthetic monoterpenes with ratios of enantiomers representative of galled and ungalled stems. PMID- 15112737 TI - Straight and branched-chain fatty acids in preorbital glands of sika deer, Cervus nippon. AB - Using GC-MS analysis, 11 major volatile compounds were found in the preorbital gland secretion from a female sika deer, Cervus nippon. These compounds are the C14 through C18 straight-chain fatty acids, (ZZ)-9,12-octadecadienoic acid, 12 methyltridecanoic acid, 13-methyltetradecanoic acid, 14-methylpentadecanoic acid, 14-methylhexadecanoic acid, and 15-methylhexadecanoic acid. The five branched chain acids make up over 29% of the volatiles in the gland. This is the first time branched-chain carboxylic acids have been reported from ungulate preorbital glands. PMID- 15112738 TI - [Impact of a guiding CT prior to angiographic intervention]. AB - With these two cases we want to demonstrate the additional impact of contrast enhanced multi-detector-row-CT angiography (MDR-CTA) compared to digital subtraction angiography (DSA) for planning of angiographic intervention. In selected cases a pre-interventional CTA can be useful to facilitate angiographic intervention. We selected two patients with different disease entities (bleeding caused by hepatic aneurysmosis; hepatocellular carcinoma (HCC) prior to transarterial chemoembolization (TACE) with aberrant arteries) from our collective who underwent CTA prior to angiographic intervention. The CT scans were performed using a 16 channel Multi-Detector-Row-CT (Philips Mx8000 IDT). Both multiplanar reconstructions (MPR) and slab maximum intensity projections (slab MIP) were performed. After CTA, patients underwent angiographic intervention (coil embolisation in the first case, TACE in the second case). MDR CTA can not only find the cause of hemorrhage but also demonstrate the exact localization of the specific vascular pathology (first case). These findings facilitate the intervention, resulting in decreased table time in the angio suite and a reduction in radiation exposure. The second case illustrates the anatomic detail achievable with MDR-CTA. Even very small aberrant arteries (crucial to the success of TACE) are revealed. These arteries did not show in overview DSA and required superselective catheterization (only performed after MDR-CTA). These cases show that MDR-CTA can provide important informations in planning of interventional procedures. PMID- 15112739 TI - [Mesenteric ossification in CT indicates sclerosing peritonitis in chronic bacterial infection and pancreatitis]. AB - Sclerosing peritonitis already has been described as a serious complication of the continuous ambulatory peritoneal dialysis. But different other affections of the peritoneum such as chronic bacterial peritonitis and pancreatitis may result in sclerosing peritonitis, too. The symptom is characterised by thickened small bowel walls and peritoneal membranes as well as peritoneal calcifications which can be shown in computed tomography. We demonstrate two cases of peritoneal ossifications due to peritonitis and pancreatitis. PMID- 15112740 TI - [Traumatic subarachnoid-pleural fistula causing tension pneumocephalus]. AB - Uncommon subarachnoid-pleural fistula can be caused by a blunt or penetrating trauma and can also occur after a thoracal or neurosurgical operation, ending to a progredient pneumocephalus. Clinical findings might be an enlarging tension pneumocephalus without a cranial fracture, but in most times they will be overlayed by other accompanied injuries, neurologic symptoms can dominate, also rapidly filling clear pleural effusions with obscure cause. Oppurtune identification of the reason of the pneumocephalus and mostly early surgical intervention prevent neurological deficit, CNS infection and other complications. PMID- 15112741 TI - [Eagle syndrome: diagnostic imaging and therapy]. AB - In the case of clinical symptoms such as dysphagia, foreign-body sensation and chronic neck or facial pain close to the ear, an Eagle syndrome should be considered in the differential diagnosis. Rational diagnostics and therapy are elucidated on the basis of four case reports. Four patients presented in the outpatients clinic with chronic complaints on chewing and a foreign-body sensation in the tonsil region. Upon specific palpation below the mandibular angle, pain radiating into the ear region intensified. In all patients, local anaesthesia with lidocaine only led to a temporary remission of symptoms. Imaging diagnostics then performed initially included cranial survey radiograms according to Clementschitsch as well as in the lateral ray path and an OPTG. An axial spiral-CT was then performed using the thin-layer technique with subsequent 3-D reconstruction. Therapy consisted of elective resection with a lateral external incision from the retromandibular. From a symptomatic point of view, the cranial survey radiograms and the OPTG revealed hypertrophic styloid processes. The geometrically corrected addition of the axial CT images produced an absolute length of 51-58 mm. The 3-D reconstruction made it possible to visualise the exact spatial orientation of the styloid processes. An ossification of the stylohyoid ligament could definitely be ruled out on the basis of the imaging procedures. After resection of the megastyloid, the patients were completely free of symptoms. Spiral-CT with subsequent 3-D reconstruction is the method of choice for exact determination of the localisation and size of a megastyloid, while cranial survey radiograms according to Clementschitsch and in the lateral ray path or an OPTG can provide initial information. The therapy of choice is considered to be resection of the megastyloid, whereby an external lateral incision has proved effective. PMID- 15112742 TI - [Health policy and occupational health: tools and methods to assure quality and appropriateness of interventions]. AB - BACKGROUND: Although all health care organizations and professionals are encouraged to follow the explicit aim of reducing the burden of illness, accidents and disability, there is increasing pressure on health professionals to ensure that the practice be based on a quality standard and on evidence of appropriateness. OBJECTIVES AND METHODS: The paper aims to focus on both the concept and dimensions of appropriateness (a complex issue with various dimensions and definitions) and on the different research and practice models aimed at search, analysis and synthesis of the best available evidence to be applied in the decision process of health and medical practice. Literature analysis has been carried out to find available tools and methods to identify, implement and monitor the evidence of efficacy in health care systems and services. RESULTS: Most definitions of appropriateness address a number of requirements. An appropriate practice should be effective (based on sound evidence); efficient (providing that cost is considered) and consistent with the ethical principles and preferences of the relevant individual, community or society. Appropriateness is considered as the next frontier in the development of health care research: there is still limited knowledge, awareness, or research on this aspect. Health Technology Assessment consists of a multidisciplinary activity based on the systematic examination of technical performance, safety, clinical effectiveness and efficiency, including socio-economic, legal and ethical aspects. Evidence Based Medicine originates from an individual perspective and aims at using rigorous criteria to obtain the best available evidence in order to provide efficient interventions. The Clinical Practice Guidelines, based on both evidence and opinions of experts, are intended to assist clinical decisions for health professional and providers for the improvement of professional practices and system efficiency, taking into account local customers, health needs and preferences. CONCLUSIONS: Several models exist for measuring and evaluating the efficacy of health systems and health services. They can be adapted for evaluating and measuring public and occupational health practices. These tools support good practice through quality-standardized interventions aiming at the health protection of communities and individuals. These models, which are presented in a specific framework in which their outcomes and critical factors are evaluated, should be adapted to public and occupational health interventions to satisfy implicit and explicit needs of individuals and populations at risk thus assuring their quality and appropriateness. PMID- 15112743 TI - [Occupational chronic obstructive pulmonary disease: Italian law (decree no. 336/1994) and epidemiological evidence]. AB - BACKGROUND: In Italy, occupational diseases are required by law to be listed (however, any occupational diseases not included in the list may be compensated if workers demonstrate causation in court). The list is periodically updated in order to incorporate new findings from medical research. Decree no. 336/94 uses a less inclusive term (chronic obstructive bronchitis) than that used in the earlier decree, no. 482/75 (chronic obstructive pulmonary disease, COPD). Moreover, it includes fewer occupations that involve a risk of developing COPD. OBJECTIVES: In order to find an epidemiological basis that might justify the above changes to Italian law, risk of COPD was investigated in relation to occupation and time, using a case-referent design. METHODS: The same definition for COPD (clinical history of >2 years with shortness of breath and/or winter phlegm and an FEV1 less than 80% of the predicted value, only minimally reversible with bronchodilators and without marked change during the hospital observation period) was used in two case-referent studies. The first study was carried out from 1972 to 1980 (202 cases of COPD and 202 referents matched for age and sex) and the second from 1990 to 1997 (131 cases of COPD and 298 referents, every fourth eligible patient in each calendar year of observation). In both studies cases and referents were classified into 16 occupational categories, and office workers were the reference. All subjects were selected from male patients admitted to the Occupational Health Clinic in Padua. Smoking adjusted (Mantel-Haenszel analysis, first study), or age-smoking-adjusted (logistic regression analysis, second study) Odds Ratio (OR) and 95% confidence interval (CI) for COPD were estimated in each occupational category and in two main groups: workers employed in occupations at either high or low risk for COPD. The latter OR and the fraction of exposed cases (pc) were used to estimate the population attributable risk (PAR) through: pc(OR-1)/OR. RESULTS: Significantly high risks for COPD in miners and chemical workers were observed in the first period, but not in the second time window. On the other hand, an increased time trend for OR estimates was found in welders, wood carpenters, construction and foundry workers. Among farmers, painters, cotton textile and refractory brick workers, the OR for COPD was significantly above unity in both time periods. PAR was 37% in the first and 52% in the second study. CONCLUSIONS: The restrictive definition of occupational COPD, which was introduced by Decree no. 336/94, contrasts with the epidemiological evidence showing that the risk of occupational COPD has increased over time: up to 50% of COPD cases referred to an occupational medicine physician might have an occupational origin. PMID- 15112744 TI - [Assessment of asbestos exposure via mineralogical analysis of bronchoalveolar lavage fluid]. AB - BACKGROUND: Mineralogical analysis of bronchoalveolar lavage fluid (BALF) by electron microscopy could be the most suitable method for assessing asbestos exposure. However, it has been claimed that there is not a standardized or systematic approach to the subject of mineralogical analysis. OBJECTIVES: The aim of the study was to evaluate mineralogical analysis of BALF by transmission electron microscopy (TEM) as biomarker of asbestos fibre load. METHODS: BALF was examined in 193 exposed workers (189 men and 4 women) and in 84 patients (65 men and 19 women) who underwent diagnostic fibreoptic bronchoscopy for various clinical purposes. Asbestos bodies (AB) in BALF were counted with a phase contrast microscope, while fibres were counted and analysed by TEM. RESULTS: Fibre counting by TEM showed a significant difference in the two populations (two tailed Mann-Whitney U test, p=0.0044), since it was positive in all exposed subjects. Only 75.1% of the exposed population was positive for asbestos bodies (AB). Subjects who had been exposed over a long time period had higher concentrations of fibres than subjects who had been exposed more recently probably because of higher exposure in the past. CONCLUSIONS: The study confirms the results of a previous study on a limited number of subjects. Fibre concentrations in BALF can be considered as a reliable biomarker of past asbestos exposure even after many years after cessation of exposure. PMID- 15112745 TI - Reference values of urinary acetone in a Brazilian population and influence of gender, age, smoking and drinking. AB - BACKGROUND: Reference values for some xenobiotics naturally present in the body are important for biomonitoring, in order to compare the levels found in a population exposed to the xenobiotic with those of a reference population. Acetone in urine (UAc) is the most used bioindicator to evaluate worker exposure to acetone and isopropanol. OBJECTIVES: Since acetone is also found in individuals not occupationally exposed to these solvents, the purpose of the present study was to evaluate the basal levels of UAc and the possible influence of individual factors on such levels. METHODS: The population consisted in 207 individuals, 91 men and 116 women, between 18 and 80 years old. UAc was determined by headspace/gas chromatography/FID. RESULTS: For the total population, the reference values found were: mean (+/- SD), 1.12 (+/- 0.47) mg/l; median 1.04 mg/l; geometric mean 1.03 mg/l; 95% confidence interval 0.98-1.26 mg/l, 95th percentile 2.20 mg/l and upper reference level (mean+2 SD) 2.06 mg/l. As the values of UAc resulted in a non-Gaussian distribution, the option was to transform these values to log UAc, which drew the data closer to normal distribution (W=0. 98532, p<0.7000). CONCLUSIONS: Reference values for acetone in urine determined in a population in south Minas Gerais, Brazil, are close to the background values reported elsewhere; gender and ingestion of alcohol seem to affect the basal levels of urinary acetone, while age or smoking showed no similar effect. PMID- 15112746 TI - [Passive smoking in bars, restaurants, and discotheques in Florence, Italy]. AB - BACKGROUND: "Environmental Tobacco Smoke (ETS) exposure in a sample of European cities" is the first European multicentre project intended to measure ETS exposure in public places in a number of European cities. OBJECTIVES: To present results of measurements of nicotine concentration in a number of bars, restaurants and discotheques in Florence, Italy. METHODS: The ETS marker was vapour-phase nicotine sampled by passive monitors. At least two monitors were placed in each of seven bars (five in hospitals; one at an airport; one at a railway station), and seven restaurants (three with smoking and non-smoking sections), and left in place for several days. In each of four discotheques two nicotine passive monitors were used as personal samplers. RESULTS: The average nicotine concentration in discotheques, restaurants and bars was respectively 26.78 micro/m3, 2.32 microg/m3 and 0.83 microg/m3. In the smoking section of restaurants with separated areas for smokers and non-smokers the average nicotine concentration was 2.54 microg/m3, which was similar to that measured in non smoking sections (2.14 microg/m3). PMID- 15112747 TI - [Estimates of pleural mesothelioma incidence in the Lazio region (Italy), 1997 2000]. AB - BACKGROUND: Malignant mesothelioma is indicative of past exposure to asbestos. In recent years an increase of incidence and mortality from malignant mesothelioma has been observed. Recent legislation in Italy requires nation-wide registration of asbestos-related pathologies. We conducted a preparatory study for systematic recording of cases of malignant pleural mesothelioma in the Lazio region. OBJECTIVES: To register new diagnoses of malignant mesothelioma, to estimate the incidence in the Lazio region, and to evaluate possible survey instruments. METHODS: We conducted a systematic study of hospital admissions in the region with diagnosis of cancer of the pleura (ICD-IX 163) in the period 1997-2000. Clinical information and results of diagnostic tests were requested for 530 patients from the hospitals involved. Using the capture-recapture method, it was possible to estimate the accuracy of the data we compiled using hospital admissions as the data source (76.8%, 95% C.I.=76.4-77.3). RESULTS: After careful review of clinical documentation, the diagnosis of malignant mesothelioma of the pleura was confirmed in 31.6% of cases (156 cases diagnosed). The percentage of confirmed cases has risen over the years (from 21% in 1997 to 45.1% in 2000) and it was higher in large public hospitals than in other types of health care facilities. On the basis of 156 confirmed cases of mesothelioma (116 males and 40 females), we estimated the annual incidence of the disease in the Lazio region as 1.73 new cases per 100,000 inhabitants among men and 0.47 new cases per 100,000 inhabitants among women. CONCLUSIONS: The results show that the incidence of mesothelioma in the region is consistent with national data, falling in the middle of the range for all Italian regions. However, some areas emerge (for example, Colleferro, Civitavecchia, Tarquinia, Ferentino, Gaeta, Aprilia, Pomezia) that have particularly high rates, probably in relation to past occupational asbestos exposure. The role of diffuse environmental exposure in Rome may warrant further investigation. PMID- 15112748 TI - Health, Environment and Social Management in Enterprises programme in the Republic of Macedonia. AB - BACKGROUND: Macedonia is the first country in the region to launch implementation of the WHO Health, Environment and Social Management in Enterprises (HESME) Programme, following the WHO Ministerial Conference on Environment and Health held in London in 1999. OBJECTIVES AND METHODS: The aim of this paper is to describe the efforts made to implement this programme. Methods are based on integrated management with joint involvement of crucial partners at all levels of activities suggested by the WHO. RESULTS: Commitment to inter-sectorial and interagency collaboration at national level, adoption of a final version of a National HESME Plan, with basic principles, criteria and concrete activities, establishment of a National coordination center for the HESME Project, development of training curricula and specific educational tools for occupational health personnel, preparation of questionnaires and procedures for a national survey to detect high occupational risks, specific occupational hazards and health promotion needs of the working population, and finally setting up quantitative and qualitative indicators for national or provincial workplace health profiles. CONCLUSIONS: Building up the concept of cooperation, partnership and common work in HESME activities is a challenge for the new public health view in Europe. PMID- 15112749 TI - [Latex allergy in health care workers: frequency, exposure quantification, efficacy of criteria used for job fitness assessment]. AB - BACKGROUND: The main purpose of this study was to evaluate the latex allergy prevalence in a large population of health care workers, to quantify latex exposure deriving from use of gloves and to verify the efficacy of job fitness evaluations in allergic workers. METHODS: In the period 2001-2002, latex allergy prevalence was evaluated in 1962 health care workers by means of a self administered questionnaire, clinical evaluation and specific allergological tests. Also, the total protein content (by means of Lowry method modified EN455 3: 1996) and the antigenic latex proteins (by means of RAST inhibition) in 4 different types of gloves were measured. Job fitness assessments for latex allergic workers were made in accordance with the criteria established by the Italian Association of Preventive Medicine for Health Care Workers criteria. The efficacy was verified one year later. RESULTS: 1557 questionnaires out of 1962 (79.35%) were completed; 504 (32.4%) reported work-related symptoms, in particular 283 subjects had probable irritant contact dermatitis, 66 allergic contact dermatitis and 118 reported irregular non-specific symptoms related to the use of both vinyl and latex gloves; 20 subjects out of the remaining 37 had a latex allergy (1.3% out of the 1557 workers responding to the questionnaire), 8 subjects had only urticaria while 12 subjects had urticaria associated with respiratory symptoms and/or angioedema. The measurement of total protein and antigenic protein content showed the highest levels in powdered latex examination gloves, the lowest levels in surgical powder-free latex gloves. Low concentrations of antigenic proteins were also found in nitrile gloves. Job fitness evaluations were efficacious in 11 out of 20 workers, and inefficacious in 3 cases (6 workers had resigned). CONCLUSIONS: This study revealed a very low prevalence of latex allergy probably due to the fact that we examined an unselected population, and only symptomatic subjects were considered; moreover, in our hospital, vinyl examination gloves had been prevalently used during the last ten years. The criteria followed for job fitness assessment seem to be efficacious on the whole. In spite of a low prevalence of latex allergy, we found a high frequency of irritant contact dermatitis in the examined workers, mainly due to the lubricant powder in both synthetic and natural rubber gloves. Most workers made a complete recovery when they started using powder-free gloves. The high biocompatibility of powder-free gloves was confirmed by the measurement of total protein and latex antigenic protein content in the gloves used in the hospital. Moreover, it should be noted that latex antigenic proteins were also demonstrated in nitrile gloves, this is a relevant information since nitrile gloves are often used as an alternative in latex allergic workers. PMID- 15112750 TI - [Health surveillance in occupational medicine: the problem of formerly exposed workers]. PMID- 15112751 TI - Assessment of upper respiratory tract and ocular irritative effects of volatile chemicals in humans. AB - Accurate assessment of upper respiratory tract and ocular irritation is critical for identifying and remedying problems related to overexposure to volatile chemicals, as well as for establishing parameters of irritation useful for regulatory purposes. This article (a) describes the basic anatomy and physiology of the human upper respiratory tract and ocular mucosae, (b) discusses how airborne chemicals induce irritative sensations, and (c) reviews practical means employed for assessing such phenomena, including psychophysical (e.g., threshold and suprathreshold perceptual measures), physiological (e.g., cardiovascular responses), electrophysiological (e.g., event-related potentials), and imaging (e.g., magnetic resonance imaging) techniques. Although traditionally animal models have been used as the first step in assessing such irritation, they are not addressed here since (a) there are numerous reviews available on this topic and (b) many rodents and rabbits are obligate nose breathers whose nasal passages differ considerably from those of humans, potentially limiting generalization of animal-based data to humans. A major goal of this compendium is to inform the reader of procedures for assessing irritation in humans and to provide information of value in the continued interpretation and development of empirical databases upon which future reasoned regulatory health decisions can be made. PMID- 15112752 TI - Physicochemical and biological data for the development of predictive organophosphorus pesticide QSARs and PBPK/PD models for human risk assessment. AB - A search of the scientific literature was carried out for physiochemical and biological data [i.e., IC50, LD50, Kp (cm/h) for percutaneous absorption, skin/water and tissue/blood partition coefficients, inhibition ki values, and metabolic parameters such as Vmax and Km] on 31 organophosphorus pesticides (OPs) to support the development of predictive quantitative structure-activity relationship (QSAR) and physiologically based pharmacokinetic and pharmacodynamic (PBPK/PD) models for human risk assessment. Except for work on parathion, chlorpyrifos, and isofenphos, very few modeling data were found on the 31 OPs of interest. The available percutaneous absorption, partition coefficients and metabolic parameters were insufficient in number to develop predictive QSAR models. Metabolic kinetic parameters (Vmax, Km) varied according to enzyme source and the manner in which the enzymes were characterized. The metabolic activity of microsomes should be based on the kinetic activity of purified or cDNA-expressed cytochrome P450s (CYPs) and the specific content of each active CYP in tissue microsomes. Similar requirements are needed to assess the activity of tissue A- and B-esterases metabolizing OPs. A limited amount of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and carboxylesterase (CaE) inhibition and recovery data were found in the literature on the 31 OPs. A program is needed to require the development of physicochemical and biological data to support risk assessment methodologies involving QSAR and PBPK/PD models. PMID- 15112753 TI - [The treatment of neoplastic colorectal occlusion: an update]. PMID- 15112754 TI - [Angiodysplasia in the gastrointestinal haemorrhage of the elderly patient. Our experience]. AB - Angiodysplasia of the digestive tract is one of the main causes of acute bleeding and is a frequent reason for admittance to the emergency surgery unit. This pathology, previously considered rare and often not recognised, has only recently acquired a precise anatomo-pathology thanks to endoscopy. Besides having a decisive diagnostic role, endoscopy also allows the control and successful treatment of lesions, often in a definitive way, which were previously only dealt with surgery, with significant advantages for the elderly patient, reducing the rate of morbidity and mortality. There are still many discussions today, above all on the priority of various diagnostic investigations to be carried out in digestive bleeding and on the choice of treatment in the case of angiodysplasic lesions in geriatric age. Some cases of acute bleeding from intestinal angiodysplasia, observed by the Authors, have led them to study the etiopathogenesis, diagnosis and emergency treatment of such lesions. PMID- 15112755 TI - [Diagnosis and treatment of distal bile duct carcinoma: our experience]. AB - Bile duct carcinomas are known to be difficult to cure, due to frequent locoregional recurrence even after radical resection. The Authors analyze their experience in a study about 53 patients treated between October 1991-January 2002. PMID- 15112756 TI - [The surgical treatment of nodular thyroid lesions: our experience. Analysis of 462 cases]. AB - Nodular thyroid lesions are the most frequent endocrine diseases in the general population. The surgical procedure is indicated for nodular thyroid tissue degeneration, or when the suppressive pharmacologic therapy is less efficient, even if sometimes both factors are associated. In Authors' study 462 patients were observed who underwent surgical procedures for thyroid diseases between January 1997-April 2003. In the thyroid pathology, either uninodular or multinodular, the surgical therapy adopted is total thyroidectomy, according to other Authors. The aim of total thyroidectomy is to avoid recurrence and simplify long term pharmacologic treatment. Although the question about the surgical approach (total thyroidectomy vs lobectomy) is still open in the case of single monolateral lesions, on the basis of their experience the Authors believe that the first is the best procedure. For diffused or malignant nodular thyroid pathology, on the contrary, total thyroidectomy is widely adopted. PMID- 15112757 TI - [Atypical presentation of angiomiolypoma in a patient with peritoneal metastases from ovarian cancer: a case report]. AB - A renal mass with not typical instrumental characteristic in patient in follow-up for ovarian neoplasia sets to the surgeon serious doubts about proper surgical strategy. Achieve of the conservative renal surgery assisted by the intraoperative use of the radiofrequency energy has allowed to preserve the renal function and the diagnosis of unknown angiomyolipoma. PMID- 15112758 TI - ['Incidentaloma' in inguinal hernia surgery]. AB - Adult Granulosa-Cell Tumor (GCT) is an uncommon ovarian malignancy. The clinical course is characterized by slow growth with late recurrence. A variety of clinical and pathologic parameters have previously been evaluated for prognosis with inconclusive results. The aim of this study was to report a case of a granulosa-cell tumor of the ovary that recurred 25 years after first surgery. The clinical course and histopathology of the case were reviewed and a literature search for other reported cases was performed. PMID- 15112759 TI - [Posterior mediastinum neoplasms: a case of schwannoma]. AB - The Authors report the case of a 33-year-old male affected with an asymptomatic schwannoma of the posterior mediastinum, and review the relevant Literature, discussing the difficulties in making the differential diagnosis of the posterior mediastinum lesions, particularly in the pre-clinical phase. Neurogenic tumors represent about 75 percent of all tumors of mediastinum and about one third of all tumors of the mediastinum. They can originate from the peripheral nervous system group, from the sympathetic nervous system group or, in rare cases, from the vagus nerve. In adult patients, they are usually found by chance during radiographic examination of the thorax, and they are usually asymptomatic and benign. During preoperative evaluation, a magnetic resonance examination of rachis should be done to exclude the possibility of intraspinal involvement. If there are no contraindications, the treatment of choice should be surgical resection by means of thoracoscopy or thoracotomy, when size and location of the tumors allow it in order to prevent malignant evolution. PMID- 15112760 TI - [Extrauterine endometriosis: three new cases]. AB - Extragonadal endometriosis is rarely diagnosed preoperatively due the variety of its localizations. Presentation to general surgeons may be atypical and pose diagnostic difficulties. The Authors report three cases surgically treated between 2000-2003. The Authors stress the peculiarity of one of these cases with endometrial tissue involvement of the intestinal mucosa. They also discuss about various aetiological hypothesis, symptomatology, differential diagnosis and need for multidisciplinary treatment. PMID- 15112761 TI - [Collateral effects of intraoperative hyperthermic chemotherapy in peritoneal carcinomatosis]. AB - The association between chemotherapy and hypertermia produces a synergic effect. In this study the Authors present their experience, by the analysis of the results. From 1993 to 2000, 17 patients have been treated with surgery associated with hypertermic chemotherapy for peritoneal carcinomatosis. For the management of these patients a constant cooperation among surgeon, cardiologist and anaesthetist is very important. PMID- 15112762 TI - [Liver hemangioma: the need for a well-defined differential diagnosis and the therapeutical timing dilemma]. AB - Liver is frequently affected by hemangiomas originated during the embryonal period from defects in the maturation of the endothelia of the capillary system. These benign tumors may result asymptomatic for the whole life (4-7% of incidental autoptical reports), but in some cases they are liable to a spontaneous or traumatic rupture, with haemorrhagic complications which make an urgent laparotomy necessary. The rupture may be caused by different reasons, such as the critical size reached by the endothelia, the development of the mass due to the continuous pulsatility or the hormonal stimuli (puberty, pregnancy or oral contraceptives). PMID- 15112763 TI - Senators unanimously approve MUMS bill. PMID- 15112764 TI - California dog owner awarded 39,000 dollars in veterinary malpractice suit. PMID- 15112765 TI - Minimizing the risk factors associated with veterinary NSAIDs. PMID- 15112766 TI - New form of BSE sparks discussion. PMID- 15112767 TI - Veterinary drug kills vultures abroad. PMID- 15112768 TI - A puppy paternity test. PMID- 15112769 TI - Wants more information on anesthesia in reptiles. PMID- 15112770 TI - Thoughts on leptospirosis vaccines. PMID- 15112771 TI - Introducing a course in veterinary medicine and literature into a veterinary curriculum. PMID- 15112772 TI - What is your diagnosis? Dystocia during a previous parturition. PMID- 15112773 TI - ECG of the month. Atrial fibrillation with a slow ventricular response rate and splintered QRS complexes. PMID- 15112774 TI - A practitioner's primer on foot-and-mouth disease. AB - Foot-and-mouth disease (FMD) is caused by an RNA virus of the genus Aphthovirus; 7 immunologically distinct serotypes of the virus have been identified. Susceptible species are mainly domestic and wild even-toed ungulates, such as cattle, sheep, goats, pigs, bison, and deer. All body fluids of infected animals can contain the virus and are considered infective. The primary mode of transmission is animal-to-animal transmission through inhalation or ingestion of aerosols containing the virus. The virus can also be spread mechanically by contaminated organic debris and fomites and can survive for 48 hours on human oral and nasal mucosa and be spread to uninfected animals in this manner. There is a rapid progression of clinical signs after an animal becomes infected, and the virus spreads rapidly throughout a herd. Clinical signs include excessive salivation; fever; vesicles and erosions of the oral and nasal mucosa, coronary band, interdigital area, and teats; lameness; sloughing of claws; reluctance to move; anorexia; mastitis; decreased milk production; and abortion or weak newborns. In mature animals, FMD has high morbidity and low mortality rates. Infected animals can become inapparent carriers of the virus. PMID- 15112775 TI - Cat scratch disease and other zoonotic Bartonella infections. PMID- 15112776 TI - Evaluation of modified Wright-staining of urine sediment as a method for accurate detection of bacteriuria in dogs. AB - OBJECTIVE: To compare the findings of light microscopic evaluation of routine unstained wet-mounted preparations and air-dried, modified Wright-stained preparations of urine sediment with results of quantitative aerobic bacteriologic culture of urine. DESIGN: Masked prospective study. SAMPLE POPULATION: 459 urine samples collected by cystocentesis from 441 dogs. PROCEDURE: Urinalyses and quantitative bacteriologic cultures of urine were performed. Unstained wet mounted preparations and air-dried, modified Wright-stained urine sediment preparations were examined by light microscopy for the presence of bacteria. RESULTS: Compared with results of quantitative bacteriologic culture, routine unstained preparations and modified Wright-stained preparations had sensitivities of 82.4% and 93.2%, specificities of 76.4% and 99.0%, positive predictive values of 40.1% and 94.5%, negative predictive values of 95.8% and 98.7%, and test efficiencies of 77.3% and 98.0%, respectively. Compared with 74 samples that yielded growth on bacteriologic culture, the routine unstained method had concordance and misclassification rates of 39.2% and 60.8%, respectively, whereas the Wright-stained method had concordance and misclassification rates of 78.4% and 21.6%, respectively. Significant associations between each of occult blood in urine, pyuria, female sex, and lower urine specific gravity with bacteriuria detected by Wright-stained sediment examination and quantitative bacteriologic culture of urine were identified. CONCLUSIONS AND CLINICAL RELEVANCE: Examination of modified Wright-stained preparations of urine sediment appeared to be a rapid, cost effective method that significantly improved the sensitivity, specificity, positive predictive value, and test efficiency of light microscopic detection of bacteriuria, compared with that of the routine unstained method. PMID- 15112778 TI - Use of a hybrid external skeletal fixator for repair of a periarticular tibial fracture in a Patagonian cavy. AB - An 8-week-old female Patagonian cavy was examined because of acute right hind limb lameness; radiography revealed a moderately displaced, comminuted fracture of the proximal third of the tibia. The fracture was stabilized with a hybrid external skeletal fixator. Two Kirschner wires were placed in the main proximal fragment, parallel to the tibial plateau and at right angles to each other. These wires were connected to a partial circular external fixator ring. Three half pins were placed in the distal fragment, and a straight connecting rod positioned on the medial side of the limb was connected to these pins and the fixator ring. A second connecting rod was positioned on the craniomedial side of the limb and was connected to the fixator ring and a fourth half pin in the distal fragment. The fracture healed without complications, and the fixator was removed 3 weeks after surgery. Hybrid external skeletal fixators combine the benefits of circular and linear external skeletal fixation methods, enabling rigid fixation of periarticular long bone fractures without adversely affecting mobility of the adjacent joint. PMID- 15112777 TI - Herbicide exposure and the risk of transitional cell carcinoma of the urinary bladder in Scottish Terriers. AB - OBJECTIVE: To determine whether exposure to lawn or garden chemicals was associated with an increased risk of transitional cell carcinoma (TCC) of the urinary bladder in Scottish Terriers. DESIGN: Case-control study. ANIMALS: 83 Scottish Terriers with TCC (cases) and 83 Scottish Terriers with other health related conditions (controls). PROCEDURE: Owners of study dogs completed a written questionnaire pertaining to exposure to lawn or garden chemicals during the year prior to diagnosis of TCC for case dogs and during a comparable period for control dogs. RESULTS: The risk of TCC was significantly increased among dogs exposed to lawns or gardens treated with both herbicides and insecticides (odds ratio [OR], 7.19) or with herbicides alone (OR, 3.62), but not among dogs exposed to lawns or gardens treated with insecticides alone (OR, 1.62), compared with dogs exposed to untreated lawns. Exposure to lawns or gardens treated with phenoxy herbicides (OR, 4.42) was associated with an increased risk of TCC, compared with exposure to untreated lawns or gardens, but exposure to lawns or gardens treated with nonphenoxy herbicides (OR, 3.49) was not significantly associated with risk of TCC. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that exposure to lawns or gardens treated with herbicides was associated with an increased risk of TCC in Scottish Terriers. Until additional studies are performed to prove or disprove a cause-and-effect relationship, owners of Scottish Terriers should minimize their dogs' access to lawns or gardens treated with phenoxy herbicides. PMID- 15112779 TI - Nodular immunocyte-derived (AL) amyloidosis in the trachea of a dog. AB - A 7-year-old castrated male Miniature Schnauzer was examined because of labored breathing and episodes of respiratory distress that progressed to collapse. On cervical radiographs, a focal soft tissue mass in the caudal cervical portion of the trachea was observed, and during tracheoscopy, a 1 x 1 cm, pedunculated, multinodular, pink, intraluminal mass extending from the dorsal tracheal membrane and obstructing approximately 80% of the tracheal lumen was seen. Tracheal resection and anastomosis was performed to remove the mass, and the dog recovered without complications. On histologic examination, the mass consisted of a large accumulation of homogeneous, faintly fibrillar eosinophilic material admixed with a predominantly plasma cell infiltrate; examination of sections stained with thioflavin T and Congo red stain confirmed that the eosinophilic material was amyloid. A diagnosis of nodular, immunocyte-derived (AL) amyloidosis was made. Seventeen months after surgery, the dog had a relapse of respiratory distress because of an extramedullary plasmacytoma involving the trachea. PMID- 15112780 TI - Use of percutaneous arterial embolization for treatment of intractable epistaxis in three dogs. AB - Intractable epistaxis can be a life-threatening condition. Surgical options are presently limited to ligation of the carotid artery. Extensive collateralization develops after occlusion of the carotid artery, so the benefits are likely only transient and the procedure cannot be repeated. In humans, endovascular treatment avoids many of the complications associated with surgery and has a lower recurrence rate than that associated with ligation of the internal maxillary artery. In 3 dogs with intractable epistaxis, embolization of the terminal portion of the maxillary artery was performed with polyvinyl alcohol particles and contrast slurry. Minor, self-limiting bleeding recurred in 1 dog 8 months after surgery; 2 dogs had no important complications or recurrence within a minimum 10-month follow-up time. Results indicate the feasibility of embolization of the terminal branches of the maxillary artery to control epistaxis in dogs that do not require surgery and as a useful adjunctive procedure prior to rhinotomy. PMID- 15112781 TI - Outcome and complications associated with treatment of pemphigus foliaceus in dogs: 43 cases (1994-2000). AB - OBJECTIVE: To identify factors affecting prognosis, outcome, and complications associated with pemphigus foliaceus in dogs. DESIGN: Retrospective study. ANIMALS: 43 dogs with pemphigus foliaceus. PROCEDURE: Medical records were reviewed for signalment, age at diagnosis, duration to diagnosis, body area affected, initial immunosuppressive regimens and concurrent use of antimicrobials and sucralfate or histamine receptor 2 blocking agent, adverse effects of treatment, duration of treatment, number of visits for follow-up care, cause of death, and credentials of the veterinarians responsible for continued care. RESULTS: The case fatality rate was 60.5%. Factors significantly correlated with survival time included concurrent use of antimicrobials during initiation of immunosuppressive treatment and a lower number of adverse effects to treatment. Treatment times lasting more than 10 months from diagnosis correlated significantly with survival. CONCLUSIONS AND CLINICAL RELEVANCE: Treatment with or prophylactic use of antimicrobials may be warranted during initial immunosuppressive treatment. The inverse correlation between survival time and number of adverse treatment effects was not unexpected because it was reflective of the owners' decision to euthanatize their dogs and of corticosteroid-related secondary diseases. Survival beyond the tenth month of treatment predicted long term survival, which suggests that dogs require careful management during the early months of treatment. PMID- 15112782 TI - Comparison of histologic lesions of endophthalmitis induced by Blastomyces dermatitidis in untreated and treated dogs: 36 cases (1986-2001). AB - OBJECTIVE: To compare prevalence of organisms and histologic changes in eyes from dogs with blastomycosis that were either untreated or undergoing treatment with itraconazole. DESIGN: Retrospective study. ANIMALS: 36 dogs with endophthalmitis associated with blastomycosis. PROCEDURE: Signalment, results of ophthalmic examination, and duration of treatment with itraconazole were extracted from medical records. Histologic sections from eyes were examined for prevalence and viability (ie, budding) of fungal organisms. A scoring system was devised to assess the degree of inflammation. RESULTS: Clinically, all eyes were blind and had signs of severe endophthalmitis. Histologically, the type and degree of inflammation and prevalence of Blastomyces dermatitidis were not significantly different between dogs treated with itraconazole and untreated dogs or among groups of dogs treated for different time periods (4 to 14, 15 to 28, or 29 to 72 days). Replication of the organisms in vascular tissues as well as avascular spaces in the eyes was similar in treated and untreated dogs. Lens rupture was seen in 12 of 29 (41%) eyes. CONCLUSIONS AND CLINICAL RELEVANCE: Persistence of inflammation in eyes of dogs with naturally occurring blastomycosis is likely attributable to the continued presence of B. dermatitidis, regardless of the duration of treatment with itraconazole. Lens capsule rupture, a common and previously unreported histologic finding, may contribute to cataract formation and continued inflammation. PMID- 15112783 TI - Type I immune-mediated polyarthritis in dogs: 39 cases (1997-2002). AB - OBJECTIVE: To determine clinical signs, laboratory findings, relationship to vaccination, and response to treatment for type I immune-mediated polyarthritis (IMPA) in dogs. DESIGN: Retrospective study. ANIMALS: 39 dogs PROCEDURE: Clinical records and radiographic reports from 3 university referral hospitals were reviewed. Clinical signs, laboratory and investigative findings, relationship to vaccination, and response to treatment were evaluated. RESULTS: Clinical signs and initial laboratory and clinical investigative findings were frequently abnormal but were nonspecific and not associated with likelihood of recovery. Time of vaccination was not associated with onset of disease. Chemotherapeutic immunosuppression resulted in complete cure in 56% of dogs. Continuous medication was required in 18% (7/39) of dogs, relapses were treated successfully in 13% (5/39) of dogs, and 15% (6/39) of dogs died or were euthanatized as a result of disease. CONCLUSIONS AND CLINICAL RELEVANCE: The possible involvement of vaccination in type I IMPA was not made clear from this study because of the small population size. Signalment, clinical signs, and results of diagnostic tests other than multiple synovial fluid analyses were generally nonspecific. Most dogs with type I IMPA responded to initial immunosuppressive treatment, but 31% (12/39) of dogs relapsed, required further treatment, or both. PMID- 15112784 TI - Energy and global warming: the great convergence. PMID- 15112785 TI - Arsenic's technology success story. PMID- 15112786 TI - Mercury trading scheme raises concerns. PMID- 15112787 TI - Monitoring water bodies with mass spectrometry. PMID- 15112788 TI - Chemical dynamics of sedimentary acid volatile sulfide. PMID- 15112789 TI - Learning as an objective within a structured risk management decision process. AB - Social learning through adaptive management holds the promise of providing the basis for better risk management over time. Yet the experience with fostering social learning through adaptive management initiatives has been mixed and would benefit from practical guidance for better implementation. This paper outlines a straightforward heuristic for fostering improved risk management decisions: specifying learning for current and future decisions as one of several explicit objectives for the decision at hand, drawing on notions of applied decision analysis. In keeping with recent guidance from two important U.S. advisory commissions, the paper first outlines a view of risk management as a policy analytic decision process involving stakeholders. Then it develops the concept of the value of learning, which broadens the more familiar notion of the value of information. After that, the concepts and steps needed to treat learning as an explicit objective in a policy decision are reviewed. The next section outlines the advantages of viewing learning as an objective, including potential benefits from the viewpoint of stakeholders, the institutions involved, and for the decision process itself. A case-study example concerning water use forfisheries and hydroelectric power in British Columbia, Canada is presented to illustrate the development of learning as an objective in an applied risk-management context. PMID- 15112790 TI - Five elements for organizational decision-making with an environmental management system. AB - Environmental management systems (EMSs) are growing in popularity as tools to manage corporate environmental issues. Despite widespread use, existing frameworks for EMSs may not provide organizations with the knowledge needed for decision-making. Through a synthesis of case studies and workshops, we suggest five elements for EMSs to be expanded for organizational decision-making. The five elements are process diagrams, long- and short-term goals linked to strategy, reliable information systems, risk assessment tools, and collaboration of environmental personnel across the organization. These five elements provide decision makers with relevant information linked to business strategy so that the organization can improve performance. The elements can be integrated with an existing EMS or used as a foundation for implementing one. PMID- 15112791 TI - Characteristics of nucleation and growth events of ultrafine particles measured in Rochester, NY. AB - Number concentrations and size distributions of particles in the size range of 0.010-0.500 microm were measured in Rochester, NY, from December 2001 to December 2002. The relationships between the number concentrations, gaseous pollutants, and meteorological parameters were examined during particle nucleation events. More than 70% of measured total number concentration was associated with ultrafine particles (UFP, 0.011-0.050 microm). Morning nucleation events typically peaking UFP number concentrations at around 08:00 were apparent in winter months with CO increases. These particles appear to be formed following direct emissions from motorvehicles during morning rush hour. There were also often observed increases in this smaller-sized range particles in the late afternoon during the afternoon rush hour, particularly in winter when the mixing heights remain lowerthan in summer. Strong afternoon nucleation events (> 30,000 cm(-3)) peaking at around 13: 00 were more likely to occur in spiring and summer months. During the prominent nucleation events, peaks of SO2 were strongly associated with the number concentrations of UFP, whereas there were no significant correlations between these events and PM2.5 and CO. Increased SO2 concentrations were observed when the wind direction was northwesterly where three SO2 sources were located. It is hypothesized that UFP formed during the events are sulfuric acid and water from the oxidation of SO2. There were also a more limited number of nucleation events followed by particle growth up to approximately 0.1 microm over periods of up to 18 h. The nucleation and growth events tended to be common in spring months especially in April. PMID- 15112792 TI - Atmospheric deposition of polycyclic aromatic hydrocarbons to Atlantic Canada: geographic and temporal distributions and trends 1980-2001. AB - Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants. The semivolatile organic compounds may disperse into the atmosphere by direct input from several sources such as the burning of fossil fuels, from motor vehicle emissions, and forest fires. Once in the atmosphere, they may travel great distances before being deposited to the earth's surface by the scavenging action of rain and snow. Up to 14 PAHs were determined in wet precipitation samples collected monthly from five sites in the four Canadian Atlantic Provinces during 1980-2001. The relatively more volatile PAHs (phenanthrene, fluoranthene, naphthalene, and pyrene) were predominant in the samples. Significant (P < 0.05) spatial variations in the deposition of some PAHs were observed among sites, but there were no consistent geographic patterns. Seasonal patterns were discernible with peak deposition for sigma6&14 PAHs occurring during the colder months of the year (December to March) and coinciding with higher energy consumption for heating and transport. The monthly volume weighed mean concentration for sigma6 PAHs has declined steadily since the mid 1980s at Kejimkujik National Park in southwest Nova Scotia, with a calculated half-life of 6.4 +/- 0.3 years. The maximum annual deposition flux of 20 microg m(-2) yr(-1) reached in 1985 for sigma6 PAHs decreased approximately 1 order of magnitude by the year 2000. The decrease in sigma6&14 PAHs for the region was found to be correlated (P < 0.05) with decreasing sulfate ion concentrations in the precipitation. The implementation of air pollution abatement programs in Canada, the United States, and elsewhere, switching to cleaner sources of energy and improved technology during the pastfew decades is most likely responsible for the observed decline. PMID- 15112793 TI - Comparing air dispersion model predictions with measured concentrations of VOCs in urban communities. AB - Air concentrations of nine volatile organic compounds were measured over 48-h periods at 23 locations in three communities in the Minneapolis-St. Paul metropolitan area. Concentrations at the same times and locations were modeled using a standard regulatory air dispersion model (ISCST3). The goal of the study was to evaluate model performance by comparing predictions with measurements using linear regression and estimates of bias. The modeling, done with mobile and area source emissions resolved to the census tract level and characterized as model area sources, represents an improvement over large-scale airtoxics modeling analyses done to date. Despite the resolved spatial scale, the model did not fully capture the spatial resolution in concentrations in an area with a sharp gradient in emissions. In a census tract with a major highway at one end of the tract (i.e., uneven distribution of emissions within the tract), model predictions atthe opposite end of the tract overestimated measured concentrations. This shortcoming was seen for pollutants emitted mainly by mobile sources (benzene, ethylbenzene, toluene, and xylenes). We suggest that major highways would be better characterized as line sources. The model also failed to fully capture the temporal variability in concentrations, which was expected since the emissions inventory comprised annual average values. Based on our evaluation metrics, model performance was best for pollutants emitted mainly from mobile sources and poorest for pollutants emitted mainlyfrom area sources. Important sources of error appeared to be the source characterization (especially location) and emissions quantification. We expect that enhancements in the emissions inventory would give the greatest improvement in results. As anticipated for a Gaussian plume model, performance was dramatically better when compared to measurements that were not matched in space or time. Despite the limitations of our analysis, we found thatthe regulatory air dispersion model was generally able to predict space and time matched 48-h average ambient concentrations of VOC species within a factor of 2 on average, results that meet regulatory model acceptance criteria. PMID- 15112794 TI - Levels of PCDD/Fs, PCBs, and PCNs in soils and vegetation in an area with chemical and petrochemical industries. AB - The concentrations of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/PCDFs), polychlorinated biphenyls (PCBs), and polychlorinated naphthalenes (PCNs) have been determined in soil and wild chard samples collected in an area of Tarragona County (Catalonia, Spain) with an important number of chemical and petrochemical industries. Samples were also collected in urban/residential zones, as well as in presumably unpolluted sites. In soils, the levels of PCDD/Fs ranged from 0.16 ng I-TEQ/kg (unpolluted sampling points) to 2.65 ng I-TEQ/kg (industrial zone), and those of sigmaPCBs ranged from 657 to 12038 ng/ kg in these same zones. In turn, sigmaPCNs ranged from 32 (unpolluted sites) to 180 ng/kg (residential/urban sites). In contrast to soil concentrations, there were not significant differences among collection zones in the levels of PCDD/Fs, PCBs, and PCNs found in chard. However, PCB and PCN concentrations in chard samples collected at the unpolluted sampling points were higher than the respective concentrations in soils. In general terms, the current concentrations of the organic pollutants analyzed in this study are similar or lower than data from previous reports in other countries. PMID- 15112795 TI - Fish, fishing, and pollutant reduction in the Baltic Sea. AB - The Baltic Sea is heavily polluted yet supports major commercial fisheries for cod (Gadus morhua), herring (Clupea harengus), and sprat (Sprattus sprattus). Emissions of persistent organic pollutants, such as polychlorinated biphenyls (PCBs) and DDT, were high during the 1960s and 1970s, and concentrations in fish and other fauna are still significant. Several models of the fluxes of these pollutants among the water, sediment, and atmosphere have been developed, but these generally omit the roles of fish and fisheries. We show that the standing stock of the most abundant fish species in the Baltic Sea was a sink for 260 kg of PCBs in the late 1980s to early 1990s and that the fishery removed as much or more PCB (31 kg yr(-1))than other budget components (e.g., degradation in the water column). Accounting for fish and fisheries could increase our understanding of the fluxes of pollutants, and banning the discard of highly contaminated organs such as cod liver could be part of the pollution management. PMID- 15112796 TI - Atmospheric mercury emissions and speciation at the sulphur bank mercury mine superfund site, Northern California. AB - One pathway for release of mercury (Hg) from naturally enriched sites is emission to the atmosphere. Elemental Hg, when emitted, will enter the global atmospheric pool. In contrast, if reactive gaseous Hg or Hg2+ (as HgCl2, HgBr2, or HgOH2) is formed, it will most likely be deposited locally. This study focused on the measurement of elemental Hg flux and reactive gaseous Hg concentrations at the Sulphur Bank Superfund Site, an area of natural Hg enrichment with anthropogenic disturbance and ongoing geothermal activity. Mean Hg emissions ranged from 14 to 11000 ng m(-2) h(-1), with the highest emissions from anthropogenically disturbed materials. Reactive gaseous Hg concentrations were the highest ever reported for a natural setting (0.3-76 ng m(-3)). Measured Hg fluxes were used within a Geographic Information System to estimate mercury releases to the atmosphere from the site. Results indicated approximately 17 kg of Hg y(-1) of is emitted to the atmosphere from the 3.8 km2 area, with half from mine waste, ore, and tailing piles and half from relatively undisturbed naturally enriched substrate. PMID- 15112797 TI - Halogen retention, organohalogens, and the role of organic matter decomposition on halogen enrichment in two Chilean peat bogs. AB - Natural formation of organohalogen compounds can be shown to occur in all natural environments. Peat bogs, which are built up exclusively of organic matter and cover approximately 3% of the total continental world area, are potentially significant reservoirs for organohalogen formation. Up to now, fluxes and retention rates of halogens and organohalogen formation in peat bogs were mostly unquantified. In our study, we investigated the retention of atmospheric derived halogens and the natural formation of organohalogens by differential halogen analysis in two peat bogs in southernmost Chile. Atmospheric wet deposition rates of chlorine, bromine, and iodine range between 600 and 36000, 6 and 160, and 1 and 3 mg m(-2) yr(-1), respectively. Mean annual net accumulation rates of these halogens in peat are calculated to be 12-72 mg of Cl m(-2), 1.7-12 mg of Br m( 2), and 0.4-1.2 mg of l m(-2). Retention rates are similarly high for iodine (36 46%) and bromine (7.5-50%), and substantially lower for chlorine (0.2-2%). To evaluate influences of peat decomposition processes on halogen enrichment, halogen concentrations were compared to carbon/nitrogen ratios (C/N). Our results indicate that up to 95% of chlorine, 91% of bromine, and 81% of iodine in peat exist in an organically bound form. The results also indicate that the concentrations of halogens, especially of bromine and iodine, in peat are largely determined by peat decomposition processes and that halogens are not conservative in bogs. PMID- 15112798 TI - Radiocarbon evidence for a naturally produced, bioaccumulating halogenated organic compound. AB - Halogenated organic compounds (HOCs) such as 1,1'-dimethyl-3,3',4,4'-tetrabromo 5,5'-dichloro-2,2'-bipyrrole (DBP-Br4Cl2) and heptachloro-1'-methyl-1,2' bipyrrole (Q1) have been detected worldwide, sometimes at high levels in Antarctic air, seabird eggs, the blubber of marine mammals, and, most notably, even human milk. To date, it has been difficult to determine whether these compounds are natural products or derived from industrial synthesis. Molecular level 14C analysis of these compounds is particularly appealing because most industrial compounds are manufactured from petrochemicals (14C-free) and natural compounds should have "modern" or "contemporary" 14C levels. To investigate the source of DBP-Br4Cl2, we isolated 600 microg of this compound (150 microg of carbon) from marine animal extracts by employing gel permeation chromatography, Florisil column chromatography, and two-dimensional preparative capillary gas chromatography. The purified DBP-Br4Cl2 was split into two samples (75 microg of carbon each) and analyzed by accelerator mass spectrometry for 14C content. The delta14C values were -449 percent per thousand and -467 percent per thousand, corresponding to conventional 14C ages of 4740 and 5000 years before present (BP), respectively. The presence of detectable 14C in the DBP-Br4Cl2 strongly points to at least a natural or biogenic source. However, these delta14C values for DBP-Br4Cl2 are more depleted than expected for a recently synthesized natural product. Several explanations are discussed, but additional samples PMID- 15112799 TI - Emissions of the refrigerants HFC-134a, HCFC-22, and CFC-12 from road traffic: results from a tunnel study (Gubrist Tunnel, Switzerland). AB - This study presents the quantification of the emissions of the refrigerants CFC 12 (CCl2F2), HCFC-22 (CHClF2), and HFC-134a (CF3CH2F) from road traffic in Switzerland. These gases are used as refrigerants in car air conditioning systems (A/C-systems) and in cool aggregates for refrigeration transport. All three substances act as greenhouse gases, and CFC-12 and HCFC-22 are in addition stratospheric ozone depleting chemicals. The measurements have been performed in a highway tunnel in the area of Zurich and cover a large number of individual vehicles, which are thought to be representative of a typical European car fleet. The average emission rates per vehicle were found to be 1.0 +/- 0.2 mg h(-1) for CFC-12, 0.6 +/- 0.4 mg h(-1) for HCFC-22, and 6.2 +/- 0.8 mg h(-1) for HFC-134a. These emission factors have been measured for driving vehicles and represent an average emission rate for all types of vehicles regardless of whether they are equipped with an A/C-unit or not. For an average vehicle equipped with an A/C unit, these results translate into losses of about 14 mg h(-1) for HFC-134a and 20-30 mg h(-1) for CFC-12, when the estimated distribution of HFC-134a-A/C-units (45%) and CFC-12-A/C-units (3-5%) in the car fleet were taken into account. The emissions of CFC-12 and HFC-134a were mainly attributed to the losses from A/C systems of passenger cars, whereas the emissions of HCFC-22 originate from losses of refrigeration systems of transport trucks. The observed emissions are discussed in respect to their environmental impact and compared to the overall greenhouse gas emissions of road traffic. PMID- 15112800 TI - Methane emissions from vehicles. AB - Methane (CH4) is an important greenhouse gas emitted by vehicles. We report results of a laboratory study of methane emissions using a standard driving cycle for 30 different cars and trucks (1995-1999 model years) from four different manufacturers. We recommend the use of an average emission factor for the U.S. on road vehicle fleet of (g of CH/g of CO2) = (15 +/- 4) x 10(-5) and estimate that the global vehicle fleet emits 0.45 +/- 0.12 Tg of CH4 yr(-1) (0.34 +/- 0.09 Tg of C yr(-1)), which represents < 0.2% of anthropogenic CH4 emissions. This estimate includes the effects of vehicle aging, cold start, and hot running emissions. The contribution of CH4 emissions from vehicles to radiative forcing of climate change is 0.3-0.4% of that of CO2 emissions from vehicles. The environmental impact of CH4 emissions from vehicles is negligible and is likely to remain so for the foreseeable future. PMID- 15112801 TI - Distribution of phthalate esters in a marine aquatic food web: comparison to polychlorinated biphenyls. AB - Dialkyl phthalate esters (DPEs) are widely used chemicals, with over 4 million tonnes being produced worldwide each year. On the basis of their octanol-water partition coefficients (Kow), which range from 10(1.61) for dimethyl phthalate to 10(9.46) for di-iso-decyl phthalate, certain phthalate esters have the potential to bioconcentrate and biomagnify in aquatic food webs. However, there are no reported field studies on the trophodynamics of phthalate ester in aquatic food webs. This study reports the distribution of 8 individual phthalate esters (i.e., dimethyl, diethyl, di-isobutyl, di-n-butyl, butylbenzyl, di(2-ethylhexyl), di-n octyl, and di-n-nonyl) and 5 commercial isomeric mixtures (i.e., di-iso-hexyl (C6), di-iso-heptyl (C7), di-iso-octyl (C8), di-iso-nonyl (C9), and di-iso-decyl (C10)) in a marine aquatic food web. DPE concentrations were determined in 18 marine species, representing approximately 4 trophic levels. Co-analysis of DPEs and 6 PCB congeners (i.e., PCB-18, 99, 118, 180, 194, and 209) in all samples produced a direct comparison of the bioaccumulation behavior of PCBs and DPEs. Lipid equivalent concentrations of the PCBs increased with increasing trophic position and stable isotope ratios (delta15N). The Food-Web Magnification Factor (FWMF) of the PCB congeners ranged from 1.8 to 9.5. Lipid equivalent concentrations of low and intermediate molecular weight DPEs (i.e., C1-C7 DPEs: dimethyl, diethyl, di-iso-butyl, di-n-butyl, benzylbutyl, and C6 and C7 isomers) did not exhibit statistically significant trends with trophic position or stable nitrogen isotope ratios (delta15N) in the food web and FWMFs were not significantly different from 1. Lipid equivalent concentrations of the high molecular-weight DPEs (i.e., C8-C10 DPEs: di(2-ethylhexyl), di-n-octyl, di-n nonyl, C8, C9, and C10) declined significantly with increasing trophic position and stable isotope ratios (delta15N), producing FWMFs between 0.25 and 0.48. These results show that all DPEs tested did not biomagnify in the studied aquatic food web whereas PCBs did biomagnify. PMID- 15112802 TI - Flux and product distribution during biological treatment of tetrachloroethene dense non-aqueous-phase liquid. AB - Flux in non-aqueous-phase liquid (NAPL)-contaminated systems containing active microbial populations (including Dehalococcoides sp.) was investigated using a quantitative mass balance and phase distribution approach. Batch systems containing mixed NAPL with an initial tetrachloroethene (PCE) mole fraction ranging from 0.1 to 0.4 provided a means for comparing systems where mass transfer and aqueous concentration were controlled by the initial NAPL composition. Although the use of mixed NAPL with increasing PCE mole fractions introduced a mass-transfer variable on the abiotic dissolution rate, it was determined that biological systems produced flux rates that were similar to each other regardless of the initial PCE mole fraction. Thus, organisms appeared to be dechlorinating near their maximum conversion rates, and the result was the accumulation of cis-1,2-dichloroethene (cDCE) followed by slow conversion to vinyl chloride (VC). Increases in the initial PCE mole fractions in the NAPL had a negative impact on product distribution due to the presence of a larger concentration of a more favorable electron acceptor. Because the mass converted to cDCE was present largely in the dissolved phase in all systems, the production of this metabolite was a favorable outcome in terms of NAPL dissolution. The pH dropped as low as 4.9 in active systems, indicating that the amount of HCl released during the reductive dechlorination process was large enough to overwhelm the buffering capacity. This pH effect was more pronounced in systems that exhibited extensive dechlorination to VC, further suggesting that rapid dechlorination of PCE NAPL can alter chemical characteristics in source zone regions. PMID- 15112803 TI - Bioavailability of PAHs: effects of soot carbon and PAH source. AB - The bioavailability of 38 individual polycyclic aromatic hydrocarbon (PAH) compounds was determined through calculation of biota-sediment-accumulation factors (BSAF). BSAF values were calculated from individual PAH concentrations in freshwater mussel, marine clam, and sediment obtained from field and laboratory bioaccumulation studies. Sediment that was amended with different types of soot carbon (SC) was used in some of the bioaccumulation experiments. BSAF values for petrogenic PAH were greater than those for pyrogenic PAH (e.g., 1.57 +/- 0.53 vs 0.25 +/- 0.23, respectively), indicating that petrogenic PAH are more bioavailable than pyrogenic PAH (p < 0.05). This trend was consistent among marine and freshwater sites. Increased SC content of sediment resulted in a linear decrease in the bioavailability of pyrogenic PAHs (r2 = 0.85). The effect of increasing SC content on petrogenic PAH was negligible. SC was considered as an additional sorptive phase when calculating BSAF values, and using PAH-SC partition coefficients from the literature, we obtained unreasonably large BSAF values for all petrogenic PAH and some pyrogenic PAH. This led us to conclude that a quantitative model to assess bioavailability through a combination of organic carbon and soot carbon sorption is not applicable among field sites with a wide range of soot carbon fractions and PAH sources, at least given our current knowledge of PAH-SC partitioning. Our data offer evidence that many factors including analysis of a full suite of PAH analytes, PAH hydrophobicity, sediment organic carbon content, sediment soot carbon content, and PAH source are importantto adequately assess PAH bioavailability in the environment. PMID- 15112804 TI - Oxygen tolerance of sulfate-reducing bacteria in activated sludge. AB - The oxygen tolerance of sulfate-reducing bacteria (SRB) present in activated sludge was studied in batch incubations using radiolabeled [35S]sulfate and a most probable number (MPN) technique employing activated sludge medium. Sulfate reduction (SR) could not be detected in activated sludge during oxic incubation or in the presence of nitrate. However, upon anoxic incubation of both freshly sampled activated sludge and activated sludge preaerated for 40 min, SR resumed immediately at an initial rate of 2 microM h(-1). During long-term aeration of activated sludge, the number of viable and culturable SRB remained constant at around 10(6) SRB mL(-1) throughout a 121 h aeration period. During the first 9 h of the 121 h aeration period, the anaerobic SR activitywas unaffected, as compared to that of an unaerated control sample, and recommenced instantaneously upon anoxic incubation. Even after 121 h of continuous aeration, SR took place within 1.5 h after anoxic incubation albeit at a rate less than 20% that of the unaerated control. As suggested by MPN estimates and the observed kinetics of SR, oxygen exposure resulted in temporary metabolic inactivation of SRB but did not cause cell death. Consequently, SRB have the potential for quick proliferation during anoxic storage of activated sludge. PMID- 15112805 TI - Evidence for the interaction of technetium colloids with humic substances by X ray absorption spectroscopy. AB - Spectroscopic extended X-ray absorption fine structure (EXAFS) evidence was obtained on the chemical environment of 99Tc(IV) atoms formed upon introduction of TcO4- into four types of laboratory-scale synthetic and natural systems which mimic in situ natural reducing conditions in humic-rich geochemical environments: (a) magnetite/pyrite in synthetic groundwater in the absence of humic substances (HSs), (b) magnetite/pyrite in natural Gorleben groundwater in the presence of HSs, (c) Boom clay sediment mixed with synthetic groundwater, and (d) Gorleben sand mixed with natural Gorleben groundwater. The investigated systems obey to pH 8-9 conditions, and all measured samples show similar EXAFS spectra for Tc, which could be fitted by a hydrated TcO2 x xH2O phase. The results are interpreted as follows: upon introduction of high concentrations (millimolar to micromolar) of TcO4-to chemically reducing environments, small Tc(IV) oxidic polymers are formed, which either may aggregate into larger units (colloids) and finally precipitate or may interact in their polymeric form with (dissolved and immobile) humic substances. This latter type of interaction--Tc(IV) colloid sorption onto HSs--differs significantly from the generally accepted metal--humate complexation and therefore offers new views on the possible reaction pathways of metals and radionuclides in humic-rich environments. PMID- 15112806 TI - Key role of the low molecular size fraction of soil humic acids for fluorescence and photoinductive activity. AB - The IHSS soil humic acid (HA) standard and two HAs from soils of very different origin (Chernozem and Ranker) were fractionated by tandem size-exclusion chromatography-polyacrylamide gel electrophoresis. From each HA, three fractions with different molecular sizes (MSs) and electrophoretic mobilities were obtained and investigated for their fluorescence properties and abilityto photoinduce the transformation of 2,4,6-trimethylphenol and herbicide fenuron. Regardless of the source of the HA, the two high MS fractions were found to be very weakly fluorescent. They photoinduced the degradations of fenuron and 2,4,6 trimethylphenol less efficiently than the bulk HA (10-50-fold and 1.4-5.3-fold, respectively). In contrast, the low MS fraction was proved to be fluorescent and to photoinduce the transformation of probes as least as efficiently than the bulk HA. These results show that (i) most of fluorophores and a great part of photoinductive chromophores are located in the low MS fractions of soil HAs and (ii) this distribution of photochemically active constituents may be characteristic across broad soil types. PMID- 15112807 TI - Mechanisms and products of surface-mediated reductive dehalogenation of carbon tetrachloride by Fe(II) on goethite. AB - Natural attenuation processes of chlorinated solvents in soils and groundwaters are increasingly considered as options to manage contaminated sites. Under anoxic conditions, reactions with ferrous iron sorbed at iron(hyro)xides may dominate the overall transformation of carbon tetrachloride (CCl4) and other chlorinated aliphatic hydrocarbons. We investigated mechanisms and product formation of CCl4 reduction by Fe(II) sorbed to goethite, which may lead to completely dehalogenated products or to chloroform (CHCl3), a toxic product which is fairly persistent under anoxic conditions. A simultaneous transfer of two electrons and cleavage of two C-Cl bonds of CCl4 would completely circumvent chloroform production. To distinguish between initial one- or two-bond cleavage, 13C-isotope fractionation of CCl4 was studied for reactions with Fe(II)/ goethite (isotopic enrichment factor epsilon = -26.5% percent per thousand) and with model systems for one C-Cl bond cleavage and either single-electron transfer (Fe(II) porphyrin, epsilon = -26.1 percent per thousand) or partial two-electron transfer (polysulfide, epsilon = -22.2 percent per thousand). These epsilon values differ significantlyfrom calculations for simultaneous cleavage of two C-Cl bonds (epsilon approximately equal to -50 percent per thousand), indicating that only one C-Cl bond is broken in the critical first step of the reaction. At pH 7, reduction of CCl4 by Fe(II)/ goethite produced approximately 33% CHCl3, 20% carbon monoxide (CO), and up to 40% formate (HCOO-). Addition of 2-propanol-d8 resulted in 33% CDCl3 and only 4% CO, indicating that both products were generated from trichloromethyl radicals (*CCl3), chloroform by reaction with hydrogen radical donors and CO by an alternative pathway likely to involve surface-bound intermediates. Hydrolysis of CO to HCOO-was surface-catalyzed by goethite butwastoo slow to account for the measured formate concentrations. Chloroform yields slightly increased with pH at constant Fe(II) sorption density, suggesting that pH-dependent surface processes direct product branching ratios. Surface-stabilized intermediates may thus facilitate abiotic mineralization of CCl4, whereas the presence of H radical donors, such as natural organic matter, enhances formation of toxic CHCl3. PMID- 15112808 TI - Uranium immobilization by sulfate-reducing biofilms. AB - Hexavalent uranium [U(VI)] was immobilized using biofilms of the sulfate-reducing bacterium (SRB) Desulfovibrio desulfuricans G20. The biofilms were grown in flat plate continuous-flow reactors using lactate as the electron donor and sulfate as the electron acceptor. U(VI)was continuously fed into the reactor for 32 weeks at a concentration of 126 microM. During this time, the soluble U(VI) was removed (between 88 and 96% of feed) from solution and immobilized in the biofilms. The dynamics of U immobilization in the sulfate-reducing biofilms were quantified by estimating: (1) microbial activity in the SRB biofilm, defined as the hydrogen sulfide (H2S) production rate and estimated from the H2S concentration profiles measured using microelectrodes across the biofilms; (2) concentration of dissolved U in the solution; and (3) the mass of U precipitated in the biofilm. Results suggest that U was immobilized in the biofilms as a result of two processes: (1) enzymatically and (2) chemically, by reacting with microbially generated H2S. Visual inspection showed that the dissolved sulfide species reacted with U(VI) to produce a black precipitate. Synchrotron-based U L3-edge X ray absorption near edge structure (XANES) spectroscopy analysis of U precipitated abiotically by sodium sulfide indicated that U(VI) had been reduced to U(IV). Selected-area electron diffraction pattern and crystallographic analysis of transmission electron microscope lattice-fringe images confirmed the structure of precipitated U as being that of uraninite. PMID- 15112809 TI - Dehalococcoides ethenogenes strain 195 reductively dechlorinates diverse chlorinated aromatic pollutants. AB - Dehalococcoides ethenogenes strain 195 dechlorinates tetrachloroethene to vinyl chloride and ethene, and its genome has been found to contain up to 17 putative dehalogenase gene homologues, suggesting diverse dehalogenation ability. We amended pure or mixed cultures containing D. ethenogenes strain 195 with 1,2,3,4 tetrachlorodibenzo-p-dioxin, 2,3,7,8-tetrachlorodibenzo-p-dioxin, 2,3 dichlorodibenzo-p-dioxin, 1,2,3,4-tetrachlorodibenzofuran, 2,3,4,5,6 pentachlorobiphenyl, 1,2,3,4-tetrachloronaphthalene, various chlorobenzenes, or a mixture of 2-, 3-, and 4-chlorophenol to determine the dehalogenation ability. D. ethenogenes strain 195 dechlorinated 1,2,3,4-tetrachlorodibenzo-p-dioxin to a mixture of 1,2,4-trichlorodibenzo-p-dioxin and 1,3-dichlorodibenzo-p-dioxin. 2,3,4,5,6- Pentachlorobiphenyl was dechlorinated to 2,3,4,6- and/or 2,3,5,6 tetrachlorobiphenyl and 2,4,6-trichlorobiphenyl. 1,2,3,4-Tetrachloronaphthalene was dechlorinated primarily to an unidentified dichloronaphthalene congener. The predominant end products from hexachlorobenzene dechlorination were 1,2,3,5 tetrachlorobenzene and 1,3,5-trichlorobenzene. We did not detect dechlorination daughter products from monochlorophenols, 2,3-dichlorodibenzo-p-dioxin or 2,3,7,8 tetrachlorodibenzo-p-dioxin. D. ethenogenes strain 195 has the ability to dechlorinate many different types of chlorinated aromatic compounds, in addition to its known chloroethene respiratory electron acceptors. Remediation of sediments contaminated with aromatic halogenated organic pollutants such as polychlorinated biphenyls and polychlorinated dibenzo-p-dioxins could require billions of dollars in the coming years. Harnessing microorganisms such as Dehalococcoides spp. that detoxify these compounds via removal of halogens may lead to cost-effective biotechnological approaches for remediation. PMID- 15112810 TI - Solvent-specific photolytic behavior of octachlorodibenzo-p-dioxin. AB - The photolysis of octachlorodibenzo-p-dioxin (OCDD) was investigated in various organic solvents under lambda > 300 nm irradiation. The rates of OCDD photolysis were highly solvent-specific. OCDD was photodegraded rapidly in toluene, n hexane, CCl4, and 1-octanol, whereas it underwent negligible photodegradation in acetonitrile, acetone, and methanol. Both OCDD photolysis and fluorescence emission did not take place in very polar solvents because polar solvent molecules efficiently quenched the excited OCDD (OCDD*). The addition of acetonitrile to an OCDD solution in toluene rapidly quenched both the fluorescence emission and the photolysis rate, which can be described by Stern Volmer analysis. The efficient photolysis in nonpolar (or less polar) solvents seems to be mediated through a charge-transfer path where the solvent and OCDD* act as an electron donor and acceptor, respectively. However, OCDD photolysis in CCl4 seems to represent the opposite case in which the solvent is an electron acceptor and OCDD* is an electron donor. Hammett sigma constants that approximately represent the electron-donating power in structurally related aromatic solvents show a good correlation with the photolysis rates. We propose that the solvent specificity in OCDD photolysis is mainly ascribed to the difference in the electron donating (or accepting) tendency among various solvents. When triethylamine that easily donates an electron to form a charge transfer exciplex with OCDD* was added, a marked enhancement in the photolysis rate was observed. PMID- 15112811 TI - Measurement of mixing-controlled reactive transport in homogeneous porous media and its prediction from conservative tracer test data. AB - Numerical and theoretical studies have indicated that pore-scale mixing can be the limiting process for reactions among dissolved compounds in porous media. It has been claimed that multicomponent reactions in porous media could be accurately estimated using mixing coefficients obtained from point-like measurements of conservative tracer concentrations. In this study, we verify these concepts experimentally by tracer tests in a homogeneously packed saturated sand column. Fiber-optic fluorometry was applied to detect point-related concentrations of fluorescein, which was used as both the conservative and the reactive tracer. In the reactive tracer experiment, an acidic solution containing the tracer was displaced by an alkaline solution without tracer. Since the fluorescence of fluorescein is quenched at low pH, the fluorescence intensity measured in the reactive breakthrough curve indicated the mixing of the two solutions. The measured reactive breakthrough curves were compared to predictions based on the conservative breakthrough curves. Predictions and measurements agreed well. Our results imply that incomplete mixing on the pore scale is of minor significance for field-scale applications. On this scale, however, even weak sorption might influence mixing significantly. PMID- 15112812 TI - In situ formed soot deposit as a carbon source for polychlorinated dibenzo-p dioxins and dibenzofurans. AB - The aim of this study was to investigate the role of in situ formed soot deposits generated during a combustion process for the formation of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDDs/Fs). In situ formed soot deposits were generated in an entrained flow reactor by using a sooting methane (CH4) flame (sooting phase), with or without chlorine doped into the flame, and fly ash added into the gas phase. The presence of fly ash in the soot deposit was found to be critical, as a catalyst for formation and/or a chlorinating agent. The presence of chlorinated aromatic structures in the soot matrix was not enough to promote de novo formation of PCDDs/Fs without the presence of fly ash. PCDFs were formed via direct release of the molecule backbone structure from the soot. PCDDs were formed via a similar mechanism as well as an equally important formation pathway of condensation reactions of C6 compounds. The formation rate of the soot/ash depositwas still at half its original activity 34 h after the deposits were formed, suggesting a persistent de novo formation occurring for a long time after the sooting incidences (memory effect). PMID- 15112813 TI - Reduction of benzene and naphthalene mass transfer from crude oils by aging induced interfacial films. AB - Semi-rigid films or skins form at the interface of crude oil and water as a result of the accumulation of asphaltene and resin fractions when the water immiscible crude oil is contacted with water for a period of time or "aged". The time varying patterns of area-independent mass transfer coefficients of two compounds, benzene and naphthalene, for dissolution from crude oil and gasoline were determined. Aqueous concentrations of the compounds were measured in the eluent from flow-through reactors, where a nondispersed oil phase and constant oil-water interfacial area were maintained. For Brent Blend crude oil and for gasoline amended with asphaltenes and resins, a rapid decrease in both benzene and naphthalene mass transfer coefficients over the first few days of aging was observed. The mass transfer coefficients of the two target solutes were reduced by up to 80% over 35 d although the equilibrium partition coefficients were unchanged. Aging of gasoline, which has negligible amounts of asphaltene and resin, did not result in a change in the solute mass transfer coefficients. The study demonstrates that formation of crude oil-water interfacial films comprised of asphaltenes and resins contribute to time-dependent decreases in rates of release of environmentally relevant solutes from crude oils and may contribute to the persistence of such solutes at crude oil-contaminated sites. It is estimated that the interfacial film has an extremely low film mass transfer coefficient in the range of 10(-6) cm/min. PMID- 15112814 TI - Bromamine decomposition kinetics in aqueous solutions. AB - The objectives of this study are to investigate the kinetics of bromamine decomposition and to identify the corresponding relevant reactions. Experiments were performed with a stopped-flow spectrophotometer system. Experimental variables investigated included pH (6.5-9.5), bromamines concentration (0.15-0.50 mM), ammonia to bromine ratio (5-100), and phosphate and carbonate buffers concentration (5-40 mM). The experimental results were consistent with a reaction scheme that involved the reversible disproportionation of monobromamine into dibromamine and ammonia (2NH2Br (k1)<=>(k(-1)) NHBr2 + NH3), followed by irreversible decomposition of monobromamine and dibromamine into products (2NHBr2 (k2) --> products and NH2Br + NHBr2 (k3) --> products). The monobromamine disproportionation reaction was found to undergo general acid catalysis, and the two subsequent decomposition reactions were found to experience base catalysis. Experimental results were analyzed for the determination of catalysis terms corresponding to H+, NH4+, H2PO4-, HCO3-, and H2O for rate constants k1 and k( 1); HPO4(2-) and H2O for k2; and OH-, CO3(2-), and H2O for k3. These constants were fitted with the Bronsted relationship, and the resulting fitting expressions were used to calculate any relevant catalysis rate constants that could not be determined at the range of experimental conditions used. PMID- 15112815 TI - Determination of strong ligand sites in sewage effluent-impacted waters by competitive ligand titration with silver. AB - A competitive ligand titration, employing Ag+, is used to determine the binding capacity of the small amounts of strong ligands (SL) in natural water samples. Strong ligands are defined here as high-affinity binding sites for group 11 and 12 metals such as Cu(I), Hg(II), and Ag(I). In addition, the conditional binding strength (log K') is determined for Ag- and SL. Diethyldithiocarbamate (DEDC) is the competitive ligand employed. The system is set at constant pH (8.1), ionic strength (0.1 M), and excess-fixed DEDC (10 microM) to determine SLs with log K' for Ag+ of >10. Silver was chosen as the titrant metal because it binds predominantly with S(-II) versus other ligands and reduced sulfur is thought to comprise the majority of SLs in natural waters. A two-phase system, water and 1,2 dichloroethane (DCE), is required due to the insolubility of Ag-DEDC in water. Added silver partitions into Ag+ and Ag-SL in the aqueous phase and into Ag-DEDC in the DCE phase. An automated system is used to add aliquots of silver and measure Ag-DEDC by UV absorbance in the DCE phase and [Ag+] by specific ion electrode in the aqueous phase. Excess addition of silver and a "Gran's" analysis gives the binding capacity of SL. The stability constant can also be determined for each addition of silver for an overall one-site SL assumption. Cysteine was used to test the method, and urban waters revealed SL capacities from about 50 to 150 nM and log K'(Ag) of 11-12. An independent analysis of chromium-reducible sulfide correlates well with the SL capacity. PMID- 15112816 TI - Multimedia fate model for hexachlorocyclohexane in Tianjin, China. AB - A level III fugacity model was applied to characterize the fate of gamma-HCH in Tianjin, China, before the 1990s when the contamination reached its maximum at steady state. Geometric means were used as model inputs. The concentrations of gamma-HCH in air, surface water, soil, sediment, crops, and fish as well as transfer fluxes across the interface between the compartments were derived under the assumption of steady state. The calculated concentrations were validated by independent data collected from the literature. There was generally good agreement between the estimated and the observed concentrations, and the differences were all less than 0.6 log units for air, water, soil, sediment, and fish and approximately 1 order of magnitude for crops. Around 97% of gamma-HCH accumulated in soil and sediment. Wastewater irrigation was not an important pathway for delivering gamma-HCH to soil as compared to the dominant source of agricultural application. Degradation and advective airflow carried much gamma HCH out of the system. Sensitivities of the model estimates to input parameters were tested, and a coefficient of variation normalized sensitivity coefficient was defined for the test. The most influential parameters were degradation rates in sediment and soil, application rates, concentrations in wastewater, and adsorption coefficients. Monte Carlo simulation was conducted for model uncertainty analysis. The model was run 20 000 times using randomly generated data from predefined log-normal distribution density functions. All calculated concentrations and fluxes were log-normally distributed. The dispersions of the calculated and observed concentrations were compared in terms of coefficients of variation to distinguish between true variability and model uncertainty. PMID- 15112817 TI - Impact of finer grid resolution on the spatial distribution of vehicle emissions inventories. AB - The ability to model air quality dispersion at increasingly smaller resolutions requires a concomitant improvement in the resolution of the gridded mobile source emissions used as input to these models. Two methods are currently used to allocate mobile emissions to grids; because of the limitations associated with these methods, their application is usually restricted to coarser grid resolutions. This paper uses a new mobile emissions inventory model (UCDrive) to explore the spatial distribution of mobile source emissions using finer grid cell resolutions. Our results indicate that the new model improves the precision of the spatial allocation of mobile source emissions, which in turn improves our ability to identify and implement pollutant control strategies. PMID- 15112818 TI - Fugacity-based indoor residential pesticide fate model. AB - Dermal and nondietary pathways are possibly important for exposure to pesticides used in residences. Limited data have been collected on pesticide concentrations in residential air and surfaces following application. Models may be useful for interpreting these data and to make predictions about concentrations in the home for other pesticides based on chemical properties. We present a dynamic mass balance compartment model based on fugacity principles. The model includes air (both gas phase and aerosols), carpet, smooth flooring, and walls as model compartments. Six size fractions of particulate matter with different fate and transport properties are included. We determine the compartmental fugacity capacity and mass-transfer rate coefficients between compartments. We compare model results to chlorpyrifos air and carpet measurements from an independent study. For a comparison, we run the same simulation for diazinon and permethrin. We quantify the effect of parameter uncertainty and model uncertainties related to the source release rate and conduct a sensitivity analysis to determine which parameters contribute most to output uncertainty. In the model comparison to chlorpyrifos measurements, the model results are of the same order of magnitude as measured values but tend to overpredict the measured data, thus indicating the need for a better understanding of emissions from treated surfaces. PMID- 15112819 TI - Propagation of uncertainty in hourly utility NOx emissions through a photochemical grid air quality model: a case study for the Charlotte, NC, modeling domain. AB - One of the major hypothesized sources of uncertainties in air quality model inputs is the emission inventory. A probabilistic hourly NOx emission inventory for 32 units of nine coal-fired power plants in the Charlotte domain for the year 1995 was propagated through the Multiscale Air Quality Simulation Platform (MAQSIP). The inventory was developed using time series techniques. Time series for a 4-d episode were simulated and propagated through the air quality model 50 times in order to represent the ranges of uncertainty in hourly emissions and predicted ozone levels. Intra-unit autocorrelation in emissions and inter-unit dependence were accounted for. The range of uncertainty in predicted ozone was greater when inter-unit dependence was included as compared to when units were treated as statistically independent. Uncertainties in maximum ozone hourly or 8 h concentrations at a specific location could be attributed to a specific power plant based upon regression analysis. Out of 3969 grid cells in the modeling domain, there were 43 and 1654 grid cells with a probability greater than 0.9 of exceeding a 1-h 120 ppb standard and an 8-h 80 ppb standard, respectively. The time series of predicted ozone values had similar autocorrelation as compared to monitored data. The implications of these results for air quality management are addressed. PMID- 15112820 TI - Selective electrochemical detection of ionic and neutral species using films of Suwannee River humic acid. AB - An electroanalytical method has been developed to investigate the uptake of redox active species by the humic acid substances. The Suwannee River humic acid (SHA) films were first cast on a glassy carbon electrode using an electrophoretic approach. The binding of a series of redox-active species to these SHA films was then probed using cyclic voltammetry at a rotating disk electrode. Neutral molecules such as hydroquinone and cationic species such as methyl viologen are able to bind with the humic membrane and exhibit high redox activity within the film. On the other hand, anionic species such as ferrocyanide are unable to attach themselves to the SHA films and thus exhibit negligible electrochemical activity. Cyclic voltammetric study of SHA films has also facilitated the determination of the partitioning constants and identification of the effect of coadsorbed ions (Ca2+) on the binding of redox species. The electroanalytical method described in this study opens up new avenues to examine the interactions and transport of charged species in a humic acid environment. PMID- 15112821 TI - Development and application of pyrolysis gas chromatography/mass spectrometry for the analysis of bound trinitrotoluene residues in soil. AB - TNT (trinitrotoluene) is a contaminant of global environmental significance, yet determining its environmental fate has posed longstanding challenges. To date, only differential extraction-based approaches have been able to determine the presence of covalently bound, reduced forms of TNT in field soils. Here, we employed thermal elution, pyrolysis, and gas chromatography/mass spectrometry (GC/MS) to distinguish between covalently bound and noncovalently bound reduced forms of TNT in soil. Model soil organic matter-based matrixes were used to develop an assay in which noncovalently bound (monomeric) aminodinitrotoluene (ADNT) and diaminonitrotoluene (DANT) were desorbed from the matrix and analyzed at a lower temperature than covalently bound forms of these same compounds. A thermal desorption technique, evolved gas analysis, was initially employed to differentiate between covalently bound and added 15N-labeled monomeric compounds. A refined thermal elution procedure, termed "double-shot analysis" (DSA), allowed a sample to be sequentially analyzed in two phases. In phase 1, all of an added 15N-labeled monomeric contaminant was eluted from the sample at relatively low temperature. In phase 2 during high-temperature pyrolysis, the remaining covalently bound contaminants were detected. DSA analysis of soil from the Louisiana Army Ammunition Plant (LAAP; approximately 5000 ppm TNT) revealed the presence of DANT, ADNT, and TNT. After scrutinizing the DSA data and comparing them to results from solvent-extracted and base/ acid-hydrolyzed LAAP soil, we concluded that the TNT was a noncovalently bound "carryover" from phase 1. Thus, the pyrolysis-GC/MS technique successfully defined covalently bound pools of ADNT and DANT in the field soil sample. PMID- 15112822 TI - Tracing atmospheric nitrate deposition in a complex semiarid ecosystem using delta17O. AB - The isotopic composition of nitrate collected from aerosols, fog, and precipitation was measured and found to have a large 17O anomaly with delta17O values ranging from 20 percent per thousand to 30% percent per thousand (delta17O = delta17O - 0.52(delta18O)). This 17O anomaly was used to trace atmospheric deposition of nitrate to a semiarid ecosystem in southern California. We demonstrate that the delta17O signal is a conserved tracer of atmospheric nitrate deposition and is a more robust indicator of N deposition relative to standard delta18O techniques. The data indicate that a substantial portion of nitrate found in the local soil, stream, and groundwater is of atmospheric origin and does not undergo biologic processing before being exported from the system. PMID- 15112823 TI - Development and application of a mobile laboratory for measuring emissions from diesel engines. 1. Regulated gaseous emissions. AB - Information about in-use emissions from diesel engines remains a critical issue for inventory development and policy design. Toward that end, we have developed and verified the first mobile laboratory that measures on-road or real-world emissions from engines at the quality level specified in the U.S. Congress Code of Federal Regulations. This unique mobile laboratory provides information on integrated and modal regulated gaseous emission rates and integrated emission rates for speciated volatile and semivolatile organic compounds and particulate matter during real-world operation. Total emissions are captured and collected from the HDD vehicle that is pulling the mobile laboratory. While primarily intended to accumulate data from HDD vehicles, it may also be used to measure emission rates from stationary diesel sources such as back-up generators. This paper describes the development of the mobile laboratory, its measurement capabilities, and the verification process and provides the first data on total capture gaseous on-road emission measurements following the California Air Resources Board (ARB) 4-mode driving cycle, the hot urban dynamometer driving schedule (UDDS), the modified 5-mode cycle, and a 53.2-mi highway chase experiment. NOx mass emission rates (g mi(-1)) for the ARB 4-mode driving cycle, the hot UDDS driving cycle, and the chase experimentwerefoundto exceed current emission factor estimates for the engine type tested by approximately 50%. It was determined that congested traffic flow as well as "off-Federal Test Procedure cycle" emissions can lead to significant increases in per mile NOx emission rates for HDD vehicles. PMID- 15112824 TI - Dry deposition fluxes and deposition velocities of trace metals in the Tokyo metropolitan area measured with a water surface sampler. AB - Dry deposition fluxes and deposition velocities (=deposition flux/atmospheric concentration) for trace metals including Hg, Cd, Cu, Mn, Pb, and Zn in the Tokyo metropolitan area were measured using an improved water surface sampler. Mercury is deposited on the water surface in both gaseous (reactive gaseous mercury, RGM) and particulate (particulate mercury, Hg(p)) forms. The results based on 1 yr observations found that dry deposition plays a significant if not dominant role in trace metal deposition in this urban area, contributing fluxes ranging from 0.46 (Cd) to 3.0 (Zn) times those of concurrent wet deposition fluxes. The deposition velocities were found to be dependent on the deposition of coarse particles larger than approximately 5 microm in diameter on the basis of model calculations. Our analysis suggests that the 84.13% diameter is a more appropriate index for each deposited metal than the 50% diameter in the assumed undersize log-normal distribution, because larger particles are responsible for the flux. The deposition velocities for trace metals other than mercury increased exponentially with an increase in their 84.13% diameters. Using this regression equation, the deposition velocities for Hg(p) were estimated from its 84.13% diameter. The deposition fluxes for Hg(p) calculated from the estimated velocities tended to be close to the mercury fluxes measured with the water surface sampler during the study periods except during summer. PMID- 15112825 TI - Environmental remediation by an integrated microwave/UV illumination technique. 8. Fate of carboxylic acids, aldehydes, alkoxycarbonyl and phenolic substrates in a microwave radiation field in the presence of TiO2 particles under UV irradiation. AB - Thermal and nonthermal effects originating when a system is subjected to a microwave radiation field in the TiO2-photocatalyzed transformation of model substances containing various functional groups (e.g., benzoic acid, phthalic acid, o-formylbenzoic acid, phthalaldehyde, succinic acid, dimethyl phthalate, diethyl phthalate, and phenol) have been examined under simultaneous irradiation by ultraviolet (UV) and microwave (MW) radiations. Characteristics of the microwave effects and the fate of each substrate during the microwave-assisted photocatalytic process were monitored by UV absorption spectroscopy, HPLC methods, total organic carbon assays, and identification of intermediates using electrospray mass spectral techniques. Microwave thermal and nonthermal effects were delineated by comparing results from MW-generated internal heat versus conventional external heating, and at constant ambient temperature under a microwave field. Factors involved in the nonthermal component of the microwave radiation were inferred for the initial adsorption of the substrate and its subsequent degradation occurring on the surface of TiO2 particles. Microwave effects bear on the mechanism through which a model substrate undergoes oxidative degradation. A characteristic feature of these effects was briefly examined by considering the behavior of polar (dipole moments) substrates in a microwave radiation field. PMID- 15112826 TI - Fate of volatile organic compounds in constructed wastewater treatment wetlands. AB - The fate of volatile organic compounds was evaluated in a wastewater-dependent constructed wetland near Phoenix, AZ, using field measurements and solute transport modeling. Numerically based volatilization rates were determined using inverse modeling techniques and hydraulic parameters established by sodium bromide tracer experiments. Theoretical volatilization rates were calculated from the two-film method incorporating physicochemical properties and environmental conditions. Additional analyses were conducted using graphically determined volatilization rates based on field measurements. Transport (with first-order removal) simulations were performed using a range of volatilization rates and were evaluated with respect to field concentrations. The inverse and two-film reactive transport simulations demonstrated excellent agreement with measured concentrations for 1,4-dichlorobenzene, tetrachloroethene, dichloromethane, and trichloromethane and fair agreement for dibromochloromethane, bromodichloromethane, and toluene. Wetland removal efficiencies from inlet to outlet ranged from 63% to 87% for target compounds. PMID- 15112827 TI - Laboratory setup for long-term monitoring of the volatilization of hazardous materials: preliminary tests of O-ethyl S-2-(N,N-diisopropylamino)ethyl methylphosphonothiolate on asphalt. AB - Contrary to commonly used pesticides, the rate of volatilization of extremely toxic chemicals such as the nerve agent O-ethyl S-2-(N,N-diisopropylamino)ethyl methylphosphonothiolate (VX) cannot be readily obtained under environmental conditions due to its high mammalian toxicity that would require extraordinary precautions. An alternative is a laboratory setup that would be used to obtain environmentally relevant data required for risk assessment studies. In this paper we describe a newly designed climatic hood that enables control of temperature, humidity, and air velocity within less than +/- 0.5% fluctuations during continuous operation. The performance of the evaporation system togetherwith the sampling and analytical procedures produced a meaningful concentration profile of vapors obtained from a 15 mg sample of VX dispersed as small droplets over a 10 x 16 cm piece of asphalt road. The released vapors amounted to approximately 30% of the applied mass, and its time course was best fitted to a triexponential curve with rate constants changing over time from 2.2 to 0.03 h(-1). The asphalt enhanced a specific degradation pathway of VX that is relatively minor in aqueous solutions. Results provide the first data on the volatilization of VX from samples of asphalt road, and offer an insight into VX behavior in the environment. PMID- 15112828 TI - Alkaline hydrolysis/polymerization of 2,4,6-trinitrotoluene: characterization of products by 13C and 15N NMR. AB - Alkaline hydrolysis has been investigated as a nonbiological procedure for the destruction of 2,4,6-trinitrotoluene (TNT) in explosives contaminated soils and munitions scrap. Nucleophilic substitutions of the nitro and methyl groups of TNT by hydroxide ion are the initial steps in the alkaline degradation of TNT. Potential applications of the technique include both in situ surface liming and ex situ alkaline treatment of contaminated soils. A number of laboratory studies have reported the formation of an uncharacterized polymeric material upon prolonged treatment of TNT in base. As part of an overall assessment of alkaline hydrolysis as a remediation technique, and to gain a better understanding of the chemical reactions underlying the hydrolysis/polymerization process, the soluble and precipitate fractions of polymeric material produced from the calcium hydroxide hydrolysis of unlabeled and 15N-labeled TNT were analyzed by elemental analysis and 13C and 15N nuclear magnetic resonance spectroscopy. Spectra indicated that reactions leading to polymerization included nucleophilic displacement of nitro groups by hydroxide ion, formation of ketone, carboxyl, alcohol, ether, and other aliphatic carbons, conversion of methyl groups to diphenyl methylene carbons, and recondensation of aromatic amines and reduced forms of nitrite, including ammonia and possibly hydroxylamine, into the polymer. Compared to the distribution of carbons in TNT as 14% sp3- and 86% sp2 hybridized, the precipitate fraction from hydrolysis of unlabeled TNT contained 33% sp3- and 67% sp2-hybridized carbons. The concentration of nitrogen in the precipitate was 64% of that in TNT. The 15N NMR spectra showed that, in addition to residual nitro groups, forms of nitrogen present in the filtrate and precipitate fractions include aminohydroquinone, primary amide, indole, imine, and azoxy, among others. Unreacted nitrite was recovered in the filtrate fraction. The toxicities and susceptibilities to microbial or chemical degradation of the polymeric materials remain unknown. PMID- 15112829 TI - Simultaneous prediction of Cryptosporidium parvum oocyst inactivation and bromate formation during ozonation of synthetic waters. AB - A model was developed to simultaneously assess Cryptosporidium parvum oocyst inactivation and bromate formation during ozonation of synthetic solutions in batch and flow-through reactors. The model incorporated 65 elementary chemical reactions involved in the decomposition of ozone and the oxidation of bromine species and their corresponding rate or equilibrium constants reported in the literature. Ozonation experiments were performed with a laboratory-scale batch reactor to evaluate the model with respect to the rate of ozone decomposition and bromate formation. The model was found to provide a good representation of experimental results when the ozone decomposition initiation reaction with hydroxide ion was assumed to produce superoxide radical instead of the alternatively proposed product hydrogen peroxide. The model was further developed to simulate the performance of a flow-through bubble-diffuser reactor with an external recirculation line. Each compartment of the reactor (bubble column and recirculation line) was assumed to behave as a plug flow reactor as supported by tracer test results, and an empirical correlation was used to represent the rate of ozone gas transfer in the bubble column. Model predictions of the performance of the flow-through ozone bubble-diffuser contactor were in good agreement with experimental results obtained for bromate formation and C. parvum oocyst inactivation under all conditions investigated. Additional model simulations revealed that hydrodynamic conditions had a more pronounced effect on C. parvum oocyst inactivation than on bromate formation. In contrast, pH had a strong effect on bromate formation without affecting the inactivation efficiency of C. parvum oocysts for a given level of exposure to ozone. These findings suggested that bromate formation could be minimized while achieving target inactivation levels for C. parvum oocysts by designing ozone reactors with hydrodynamic conditions approaching that of an ideal plug flow reactor and by lowering the pH of the target water. PMID- 15112830 TI - Oxidative degradation of the carbothioate herbicide, molinate, using nanoscale zero-valent iron. AB - Degradation of the carbothiolate herbicide, molinate, has been investigated in oxic solutions containing nanoscale zero-valent iron particles and found to be effectively degraded by an oxidative pathway. Both ferrous iron and superoxide (or, at pH < 4.8, hydroperoxy) radicals appearto be generated on corrosion of the zero-valent iron with resultant production of strongly oxidizing entities capable of degrading the trace contaminant. PMID- 15112831 TI - Selective separation of La3+ and lanthanum organic complexes with nanometer-sized titanium dioxide and their detection by using fluorination-assisted electrothermal vaporization ICP-AES with in-situ matrix removal. AB - A new method for the determination of free La3+ and La organic complexes in solution using a nanometer-sized titanium dioxide as solid-phase extractant and fluorination-assisted electrothermal vaporization (FETV)-ICP-AES as sensitive detector has been developed. The effect of pH on the adsorption characteristics of La3+ and La complexes of citric acid, 2-hydroxyisobutyric acid (HIBA), and humic acid on nanometer-sized TiO2 was investigated and optimized. On the basis of the difference in volatility between fluoride of analyte (lanthanum) and the fluoride of matrix (titanium), an in-situ removal of the adsorbent matrix (TiO2) from a graphite furnace was realized. Therefore, the free La3+ and adsorbed La complexes on nanometer-sized titanium dioxide could be determined respectively by FETV-ICP-AES without any other chemical pretreatment. The proposed method was applied for the determination of free ion (La3+) and La complexes in synthetic solutions and soil extracts with satisfactory results. PMID- 15112832 TI - Partitioning of metal species during an enriched fuel combustion experiment. speciation in the gaseous and particulate phases. AB - Combustion processes are the most important source of metal in the atmosphere and need to be better understood to improve flue gas treatment and health impact studies. This combustion experiment was designed to study metal partitioning and metal speciation in the gaseous and particulate phases. A light fuel oil was enriched with 15 organometallic compounds of the following elements: Pb, Hg, As, Cu, Zn, Cd, Se, Sn, Mn, V, Tl, Ni, Co, Cr, and Sb. The resulting mixture was burnt in a pilot-scale fuel combustion boiler under controlled conditions. After filtration of the particles, the gaseous species were sampled in the stack through a heated sampling tube simultaneously by standardized washing bottles based sampling techniques and cryogenically. The cryogenic samples were collected at -80 degrees C for further speciation analysis by LT/GC-ICPMS. Three species of selenium and two of mercury were evidenced as volatile species in the flue gas. Thermodynamic predictions and experiments suggest the following volatile metal species to be present in the flue gas: H2Se, CSSe, CSe2, SeCl2, Hg(0), and HgCl2. Quantification of volatile metal species in comparison between cryogenic techniques and the washing bottles-based sampling method is also discussed. Concerning metal partitioning, the results indicated that under these conditions, at least 60% (by weight) of the elements Pb, Sn, Cu, Co, Tl, Mn, V, Cr, Ni, Zn, Cd, and Sb mixed to the fuel were found in the particulate matter. For As and Se, 37 and 17%, respectively, were detected in the particles, and no particulate mercury was found. Direct metal speciation in particles was performed by XPS allowing the determination of the oxidation state of the following elements: Sb(V), Tl(III), Mn(IV), Cd(II), Zn(II), Cr(III), Ni(II), Co(II), V(V), and Cu(II). Water soluble species of inorganic Cr, As, and Se in particulate matter were determined by HPLC/ICP-MS and identified in the oxidation state Cr(III), As(V), and Se(IV). PMID- 15112833 TI - A polymer membrane containing Fe(0) as a contaminant barrier. AB - A poly(vinyl alcohol) (PVA) membrane containing iron (Fe(0)) particles was developed and tested as a model barrier for contaminant containment. Carbon tetrachloride, copper (Cu2+), nitrobenzene, 4-nitroacetophenone, and chromate (Cr04(2-)) were selected as model contaminants. Compared with a pure PVA membrane, the Fe(0)/PVA membrane can increase the breakthrough lag time for Cu2+ and carbon tetrachloride by more than 100-fold. The increase in the lag time was smaller for nitrobenzene and 4-nitroacetophenone, which stoichiometrically require more iron and for which the PVA membrane has a higher permeability. The effect of Fe(0) was even smaller for CrO4(2-) because of its slow reaction. Forty five percent of the iron, based on the content in the dry membrane prior to hydration, was consumed by reaction with Cu2+ and 15% by reaction with carbon tetrachloride. Similarly, 25%, 17%, and 6% of the iron was consumed by nitrobenzene, 4-nitroacetophenone, and CrO4(2-), respectively. These percentages approximately double when the loss of iron during membrane hydration is considered. The permeability of the Fe(0)/PVA membrane after breakthrough was within a factor of 3 for that of pure PVA, consistent with theory. These results suggest that polymer membranes with embedded Fe(0) have potential as practical contaminant barriers. PMID- 15112834 TI - Degradation of waste gas containing toluene in an airlift bioreactor. AB - Suspension microorganisms in an internal-loop airlift bioreactor were utilized to treat waste gas containing toluene. The working volume of the reactor was 35 L, and the biomass concentration was 3 kg/m3. The gas pollutant flowed into the reactor from the bottom; it then transferred from the gas phase to the liquid phase and was degraded by the microorganisms suspended in the liquid phase. The microorganisms were able to degrade 50-90% of the inlet toluene when its concentration was from 0.5 to 10 g/m3, and the superficial gas velocity ranged from 0.15 to 1.23 cm/s. A comprehensive mathematical model was also developed to describe the overall degradation process of toluene in the internal-loop airlift bioreactor. The overall degradation process included gas flow, gas-liquid mass transfer, flow and dispersion of the liquid phase, and microbial kinetics. The hydrodynamic properties including the gas and liquid superficial velocities, the gas holdup, the volumetric mass transfer coefficients of toluene and oxygen, and the microbial kinetics were obtained for this model. The substrate inhibition theory was used to simulate the microorganism growth kinetics, and its kinetic constants were obtained experimentally. The penetration theory was used to predict the volumetric mass transfer coefficient. And the drift-flux theory was used to predict the hydrodynamic properties in each section (riser, gas-liquid separator, downcomer, and bottom) of the airlift bioreactor. The concentration distributions of toluene and oxygen in the airlift bioreactor and the removal efficiency of toluene predicted by the mathematical model agreed well with the experimental data. PMID- 15112835 TI - Production of electricity during wastewater treatment using a single chamber microbial fuel cell. AB - Microbial fuel cells (MFCs) have been used to produce electricity from different compounds, including acetate, lactate, and glucose. We demonstrate here that it is also possible to produce electricity in a MFC from domestic wastewater, while atthe same time accomplishing biological wastewater treatment (removal of chemical oxygen demand; COD). Tests were conducted using a single chamber microbial fuel cell (SCMFC) containing eight graphite electrodes (anodes) and a single air cathode. The system was operated under continuous flow conditions with primary clarifier effluent obtained from a local wastewater treatment plant. The prototype SCMFC reactor generated electrical power (maximum of 26 mW m(-2)) while removing up to 80% of the COD of the wastewater. Power output was proportional to the hydraulic retention time over a range of 3-33 h and to the influent wastewater strength over a range of 50-220 mg/L of COD. Current generation was controlled primarily by the efficiency of the cathode. Optimal cathode performance was obtained by allowing passive air flow rather than forced air flow (4.5-5.5 L/min). The Coulombic efficiency of the system, based on COD removal and current generation, was < 12% indicating a substantial fraction of the organic matter was lost without current generation. Bioreactors based on power generation in MFCs may represent a completely new approach to wastewater treatment. If power generation in these systems can be increased, MFC technology may provide a new method to offset wastewater treatment plant operating costs, making advanced wastewater treatment more affordable for both developing and industrialized nations. PMID- 15112837 TI - Hearing and the brain: audiological consequences of neurological disorders. PMID- 15112836 TI - Comment on "high-resolution gas chromatography retention data as basis for the estimation of Kow values using PCB congeners as secondary standards". PMID- 15112838 TI - Central auditory deficits associated with compromise of the primary auditory cortex. AB - The subject of this study was a 46-year-old female who had suffered a cerebrovascular accident (CVA). Magnetic resonance imaging revealed damage in the area of the distribution of the middle cerebral artery involving most, if not all, of the primary auditory area of the left hemisphere. No auditory problems were noted prior to the CVA; however, following the CVA, the subject reported a number of auditory difficulties. Pure-tone thresholds were normal post-CVA, and performance on speech recognition testing was good in both ears if ample time was provided between a response and the presentation of the next test item. Duration pattern, intensity discrimination, and middle latency response test results were abnormal for both ears, and right ear deficits were evident on an auditory fusion test and two dichotic speech tests (digits and rhymes). This case is significant in that it demonstrates a good correlation between damage to known key auditory regions and central auditory test results. PMID- 15112839 TI - Assessment and remediation of an auditory processing disorder associated with head trauma. AB - This case study involves a 41-year-old female who had sustained a mild traumatic brain injury during a horseback riding accident. The patient was seen for medical and neuropsychological testing following this incident and was referred to a speech-language pathologist for rehabilitative services. At 13 months posttrauma, the patient, who was frustrated by a lack of significant progress, requested an audiologic work-up. Results of testing conducted at this time revealed normal peripheral hearing and significant central auditory deficits. Based on these findings, an auditory rehabilitation program was developed and implemented. The components of this patient's rehabilitation program are reviewed, and the posttherapy improvements noted in her auditory functions are detailed. The case is important in that it demonstrates (1) that auditory deficits can be a sequel to minor head injury, (2) that these deficits are often subtle and may not be detected unless central auditory testing is conducted, and (3) that these deficits may be amenable to remediation. PMID- 15112840 TI - Central deafness associated with a midbrain lesion. AB - Central deafness has been linked historically to bihemispheric involvement of the temporal lobe, with more recent findings suggesting that compromise of other cortical and subcortical structures can also result in this disorder. The present investigation extends our understanding of the potential anatomical correlates to central deafness by demonstrating that bilateral involvement of an auditory structure within the midbrain can additionally result in this condition. Our subject was a 21-year-old male with a subarachnoid bleed affecting both inferior colliculi. Significant auditory deficits were noted for the middle and late auditory evoked potentials, while electrophysiologic measures of the periphery indicated normal function. The patient was enrolled in a rehabilitation program for approximately 14 weeks. Although initially unresponsive to sounds, the patient regained most of his auditory abilities during the 10 months he was followed. This case documents the range of auditory deficits that may be associated with damage to the inferior colliculi, and it profiles a hierarchical recovery of auditory function consistent with test findings. PMID- 15112841 TI - An unusual case of X-linked adrenoleukodystrophy with auditory processing difficulties as the first and sole clinical manifestation. AB - X-linked adrenoleukodystrophy (X-ALD) is characterized by demyelination that is associated with a deficient beta-oxidation of very long chain fatty acids. We report the unusual case of a male adult with X-ALD who was diagnosed at the age of 26 by a brain MRI performed because his brother had been diagnosed with a rapidly deteriorating form of X-ALD. His sole symptom was hearing difficulties in the presence of a normal audiogram since childhood. He has remained stable for seven years. Central auditory testing in our patient revealed severe deficits in several auditory processes. These findings correlated with involvement of the auditory pathway at the level of the trapezoid body, and posterior corpus callosum in particular, on his brain MRI. This case highlights not only the need for thorough audiological investigation of the patient who complains of hearing difficulties in the presence of a normal audiogram, but also that audiological investigations could be of value in the phenotypic evaluation of cases with adrenoleukodystrophy. PMID- 15112842 TI - Audiological correlates to a rupture of a pontine arteriovenous malformation. AB - This is a report of a female patient in her midthirties who sustained a hemorrhage secondary to an arteriovenous malformation in the region of the pons. The patient's initial symptoms included hearing loss and tinnitus, which were followed by the more characteristic symptoms of headache and loss of consciousness. Results of audiological testing at three months postaccident documented the presence of a hearing loss and a central auditory processing disorder, and the patient was provided an auditory rehabilitation program. Follow up testing over the course of an additional year documented improvement in both pure-tone threshold and central test results; however, at 15 months postaccident, some auditory deficits remained, especially in the ear ipsilateral to the primary site of lesion. The anatomical correlates of these deficits are discussed, as are the potential contributions of both the auditory rehabilitation program and spontaneous recovery mechanisms to the documented improvements in auditory function. PMID- 15112843 TI - [The enhancement of development at nursery school age]. AB - A scientifically based synthesis of diagnostic inventaries and intervention measures would be desirable for the practice of the enhancement of development. Recent research on preschool training, however, has hitherto tended to deal with the development and testing of the effectiveness of training programmes without linking them explicitly to a diagnosis of the abilities which are to be promoted. This article presents two studies that are part of a major project to connect the assessment and enhancement of development. The objective was to develop and empirically test intervention measures for two functional areas of the Wiener Entwicklungstest (WET, Kastner-Koller and Deimann 1998, 2002). Two training conditions were developed for the area of language, i.e. a specific encouragement of vocabulary and conceptualisation and general linguistic promotion. Similarly, two different promotion strategies were realized for the functional area Visual Perception, i.e. a conventional means of promotion using work sheets and a form of promotion utilizing games. According to area, these treatments were tested for their differential effectiveness in an experimental design (linguistic training N = 42; training of perception N = 51), using the Wiener Entwicklungstest (WET) for the pre- and posttests, the intermediate programme comprising 16 units and a control group meantime taking part in the educational programme of the nursery school in both studies. The results showed that both forms of linguistic training and the promotion of perception through games were highly effective in their areas, but that the effect of work sheets could not be definitively assessed. The different effects of the training programmes on children with and without a general developmental lag in the Wiener Entwicklungstest (WET) were particularly interesting. PMID- 15112844 TI - [Co-production of knowledge about custody criteria with subjects affected by their parents' custody dispute: a participatory study]. AB - This article explores methods within a longitudinal study for allowing subjects more direct participation in research as co-producers of scientific knowledge concerning custody decision-making after divorce. The purpose of the longitudinal study was to evaluate the scientific custody criteria that were applied to children after their parents' divorce. The results of the study, published after the first survey, showed the relevance of children's personal preferences and residence wishes for custody regulations. This was formulated as a general rule for custody decision-making. In the second survey, a copy of our scientific publication written after the first survey, including summaries and a questionnaire was sent to the children involved. They were asked to judge the presentation of the custody problem, the custody recommendations and the presentation of their case history in our publication. Most of the participants (60%) voted for the application of the custody criterion "personal relationship preferences and residence wishes", but they also pointed out the difficulties for the child to articulate those wishes. Together with other studies, the participatory study contributed to a paradigm shift: from an understanding of the custody problem as a structural question concerning the suitability of maternal, paternal or joint custody to a conceptualisation of the custody problem as a decision-making process that demands the participation of the child and professional support if need be. Finally, the value of participatory research methods in divorce research and longitudinal social studies of children and childhood are discussed. PMID- 15112845 TI - [Obesity and coping in childhood]. AB - The aim of this study was to investigate everyday coping styles in obese children in comparison to healthy controls in dependence of gender. Furthermore, the correlations of eating behavior with coping styles were examined. In addition, groups with highest and lowest extremal scores in eating behavior were compared. Patients (N = 58) aged from 11 to 13 years, who took part in an in-patient education program, were asked to complete the German Coping Questionnaire for Children and Adolescents (Hampel et al. 2001) and the Dutch Eating Behavior Questionnaire for Children (Franzen u. Florin 1997). Healthy controls were matched by age and gender and were asked to fill out the SVF-KJ. No differences between coping styles of obese children and adolescents and healthy controls were found. In obese children and adolescents, emotional and external eating behavior were positively correlated with maladaptive coping strategies. In comparison to previous findings in chronically ill children, differences in obese children and adolescents and healthy controls on coping styles could not be confirmed in the present study. However, results support that the eating behavior of obese children and adolescents is influenced by coping styles. PMID- 15112846 TI - [Separation of parents: how is it conveyed to children? How does the content of the communication influence the separation experience of children?]. AB - On the basis of 45 expert opinions of custody cases involving 89 children dating from 1996-1999 and of 62 questionnaires answered by the parents it was examined, how the parents told their children about the impending separation. We were interested if the way the parents told their children and the contents of the communication had an impact on coping with the separation. We found that only 48.9% of the families talked about the separation. 35.5% of the children were not informed, 12.4% of the children were not told the truth by their parents. There was no clear connection between the way the children were informed and their coping behaviour. Yet it could be shown that talking to the children had a positive effect on the relationship between the father and the child. The discussion of the results will point out the complexity and importance of this topic which has not been sufficiently considered in the literature and by parents planning a divorce. Yet looking at the rising divorce rate one can see how highly topical it is and will remain in the future. PMID- 15112847 TI - [Rehabilitation of thoracic outlet syndrome: about 60 patients]. AB - Sixty patients were treated by physical therapy for thoracic outlet syndrome (TOS) between 1995 and 1999. Females were predominant (76%), the average age was 34.5 years (18 to 60). Risk factors were: hypermobility, cervical traumatism and profession. Neurological signs were predominant and positive provocation tests were found: Adson (48%), sonnette (62%), E.A.S.T. (51%). Thoracic pain inaugurated the symptoms in 3 times: pseudo-angina (2), dyspnea (1). Radiological results were positive for: cervical rib (8), apophysomegaly of C7 (25), clavicular traumatism (1). Electromyogram was positive in 2 out of 3 patients, RMI in 2 of 4, and dynamic echo Doppler in 3 of 4. After rehabilitation, immediate results were satisfactory in more than 70% of cases. After six months one patient had recurrence of symptoms, 66% were compliant with treatment, and 84% were satisfied. At one year the symptoms recurred in 2 patients, 15% were still compliant, and 81% were satisfied. Surgery was needed in 4 patients with cervical ribs. PMID- 15112848 TI - [Transcatheter closure of the patent ductus arteriosus in children. The first Lebanese series]. AB - Percutaneous closure of the patent ductus arteriosus (PDA) has become an efficient and safe technique in children. We report the first Lebanese series of 8 children who had their PDA closed percutaneously with a coil or an Amplatzer Duct Occluder. METHODS: Patients median age was 3.2 years (range 1.2 to 12 years). The median minimal diameter of the PDA was 2.1 mm (1.2 to 4 mm). A coil was used in 4 children and an Amplatzer occluder in the 4 other patients. The device was chosen according to the diameter of the PDA. RESULTS: The results were excellent in all patients. A trivial shunt present in 3 cases, disappeared within 8 weeks. No complication were noted in this series. CONCLUSION: Percutaneous closure of the PDA in children, using interventional radiology, is a simple and reproducible technique. It is now the treatment of choice in most pediatric cardiac centers worldwide. PMID- 15112849 TI - [Laparoscopic adrenalectomy. Results of 10 cases]. AB - AIM OF THE STUDY: To report our experience in laparoscopic adrenalectomy and to evaluate the effectiveness of this technique for a variety of endocrine disorders. METHODS: Ten patients were operated for laparoscopic adrenalectomy between January 1998 and January 2001. RESULTS: Eight females and two men, of 45 years mean age (range, 16-71 years), were operated for laparoscopic adrenalectomy. There were 8 tumours in right adrenal gland and two in the left adrenal gland. Indications were: pheochromocytomas (n = 5), aldosterone-producing adenoma (n = 2), cortisol-producing adenoma (n = 2) and cyst (n = 1). All patients had a CT Scan or RMI preoperatively. The patients with pheochromocytoma were prepared preoperatively. Mean operative time was 133 minutes (range, 110-190 minutes). There was no mortality and morbidity was encountered in one patient (bleeding in a trocart site peroperatively). During pheochromocytoma operation, hypertension occurred in two patients. There was one conversion secondary to bleeding. The average length of stay was 3.6 days (range, 2-7 days). During follow-up (range, 6-36 months), the patients were satisfied and none had recurrence of hormonal excess. CONCLUSION: Laparoscopic adrenalectomy is safe, effective and decreases hospital stay and convalescence. The laparoscopic approach is the procedure of choice for adrenalectomy except in the case of carcinoma. PMID- 15112850 TI - [Regional anesthesia for lumbar microdiscectomy]. AB - GOAL OF THE STUDY: Lumbar microdiscectomy surgery is already performed under spinal anesthesia (SA) in many institutions. The aim of this study is to compare the quality of analgesia and recovery after SA when compared to general anesthesia (GA) after lumbar microdiscectomy surgery. METHODS: Following light sedation, SA is performed with the patient in the left lateral decubitus position, one to two levels above the herniated disc level. Isobaric 0.5% bupivacaine 3-3.5 ml was injected intrathecally followed by wound infiltration with 15 ml of bupivacaine with 1/200 000 epinephrine prior to surgical incision. RESULTS: Despite randomization, we found significantly more females in the GA group. Pain scores at 4 and 8 h postoperatively were lower in SA group as well as total analgesic consumption during the first 24 h. Postoperative recovery including time to drinking, eating and walking were more rapid after SA when compared to GA. During the postoperative period, the incidence of urinary retention was comparable between groups but the occurrence of postoperative nausea and vomiting was significantly higher in the GA group. Moreover, the overall patient's and surgeon's satisfaction were significantly better in the SA group. CONCLUSION: SA associated to wound infiltration using bupivacaine is an interesting alternative to general anesthesia for outpatient lumbar microdiscectomy surgery. PMID- 15112851 TI - Survey of antimicrobial prophylaxis for surgical procedures in Lebanese hospitals. AB - Antimicrobial prophylaxis (AMP) is an effective measure to prevent the surgical site infection (SSI). This prospective survey of three months describe the adequacy of AMP in 10 acute care hospitals (affiliated to the Lebanese University, Faculty of Medical Sciences) based on international guidelines for only three parameters, digestive, orthopaedic and vascular surgery. Out of 964 surgical procedures, 916 (95%) were eligible for AMP, of which 767 (84%) received one or several antibiotics. Cefazolin was frequently prescribed (49%), followed by beta-lactamase-inhibitor/penicillin (18%), cefuroxime (17%), ceftriaxone (7%) and vancomycin (0.1%). In compliance with current US guidelines on indication, choice of drug, duration and time of first dose administration, AMP was given for only 32% of the procedures. Duration of AMP was < or = 24 hours in 35% and exceeded 48 hours in 57%. A single dose of prophylaxis was given to 26%, 12% receive an additional dose and 62% receive more than 2 doses. The first dose was administered within 30 min before operation in 92% of procedures. We conclude that AMP in Lebanese hospitals was fairly adequate in terms of respecting indications for selected surgical procedures. Improvement could be made by reducing the duration of prophylaxis and avoiding the use of broad-spectrum antibiotics. PMID- 15112852 TI - Medication use, gender, and socio-economic status in Lebanon: analysis of a national survey. AB - From data collected in Lebanon by the 1999 National Household Health Expenditures and Utilization Survey an analysis was made of reported medication use (n = 14,142). Seventy-two percent of the population aged 25-64 reported taking a medication in the month preceding the survey. Medications with high frequencies of reported use were analgesics, psychotropics, antibiotics and vitamins. Women were more than one and a half times more likely than men to report taking any medication (OR = 1.6), and significantly more likely to report taking eight of the 19 medication types analyzed. Analyses of the association of medication use with socio-economic variables show different effects for education and employment (inversely correlated with higher use), and higher socio-economic status (positively correlated with higher use). Multivariate analyses were carried out on two of the frequently used medications, antibiotic and prescription psychotropic use, to further explore the simultaneous effects of socio-economic variables The results of the analysis high-light a number of potential areas which may be targeted for intervention, in particular the higher use of antibiotics in rural areas, the greater use of psychotropics by women, and the possible obstacles to obtaining needed medications for those with lower incomes. PMID- 15112853 TI - [Gastric stromal tumors. Presentation of two cases with immunohistochemical study. Review of the literature]. AB - GSTs are uncommon mesenchymal tumors of the stomach. The evaluation of malignancy is difficult and their histogenesis is controversial. Recently a relationship to the pacemaker interstitial cells of Cajal (ICC) has been suggested because GSTs, like the ICC express CD117 (C-KIT protein). Some authors, like Miettinen in one of his studies, consider the CD117 positivity as part of the definition of GSTs. The purpose of this paper is to present two cases of GST with immunohistochemical study confirming the diagnosis of GST; the immunoreactivity of these two tumors is positive for CD117 and negative for protein S-100, actine and desmin. A review of the clinicopathologic, the immunohistochemical features, the prognostic factors and novelties in treatment will be presented. PMID- 15112854 TI - Imaging of common benign solid liver tumors. AB - The imaging modalities currently used for radiologic detection and characterization of hepatic neoplasms include color Doppler sonography, helical computed tomography (plain, biphasic enhanced CT), magnetic resonance imaging (plain, gadolinium enhanced MR), radionuclide scintigraphy (Technetium RBC, sulfur-colloid, IDA scan), angiography, and image-guided percutaneous needle biopsy. MR imaging is probably better at characterizing lesions than CT scan, but the latter remains the modality of choice at many institutions due to the speed of acquisition and good contrast resolution inherent in the technique. Each of these modalities is useful, but accurate lesion detection and specific radiologic diagnosis is most often made in many patients with typical imaging features using a combination of examinations rather than with a single modality. In the remaining patients with atypical features, a definitive diagnosis is usually obtained by a needle biopsy. Table I summarizes the typical clinical and imaging findings of common benign solid liver tumors, with suggested further work-up and management. Future developments will focus on the use of special contrast agents in sonography and MRI for further evaluation and characterization of focal hepatic lesions. Emerging clinical applications of these special contrast agents have shown a promising future role for contrast-enhanced sonography and MRI as problem-solving examinations after inconclusive ultrasound and helical CT. PMID- 15112855 TI - Immune hemolytic anemia following renal transplantation: description and guidelines. PMID- 15112856 TI - Vitamin B12 deficiency among elderly patients an important diagnosis. PMID- 15112857 TI - Perinatal factors and children's bone health. PMID- 15112858 TI - Primary actinomycosis of thigh presenting with soft tissue mass. PMID- 15112859 TI - Determination of gabapentin-lactam in serum of patients under gabapentin therapy. AB - Gabapentin (1-(aminomethyl)cyclohexane acetic acid, CAS 601 42-96-3, GBP, Neurontin) and its derivative gabapentin-lactam (8-aza-spiro[5,4]decan-9-one, GBP L) were determined by HPLC in the serum of patients with focal epilepsy treated with GBP. In patients in whom serum was acquired within 3 h after oral intake, GBP-L could be detected at concentrations up to 8.2 micromol/l. As GBP-L has been previously shown to exert neuroprotective effects in a similar concentration range, this finding suggests that clinically relevant effects of GBP-L may occur in patients treated with GBP. The possible neuroprotective efficacy of GBP-L should be the subject of further preclinical and clinical investigations. PMID- 15112860 TI - Dissociation of morphine analgesia and sedation evaluated by EEG measures in healthy volunteers. AB - AIM: The analgesic effects of morphine (CAS 57-27-2) in clinical use are well described. Sedation is discussed as a relevant side-effect, mostly based on data recorded in normal subjects without pain. The aim of this study was to quantify and to evaluate electrophysiologically the analgesic and sedative effects of morphine for the first time using an experimental pain model. METHODS: Analgesic and sedative effects of a low dose of morphine sulfate (CAS 6211-15-0; 10 mg i.v.) were determined using a standard phasic pain model (intracutaneously administered electrical pulses) in a placebo-controlled design with seven healthy subjects. Five blocks (1 block = 80 stimuli) of painful stimuli were applied, covering a period of 3 h. Analgesia was assessed by subjective pain ratings and by pain-related brain potentials. Sedation was determined by the power spectra of the spontaneous EEG, by auditory evoked potentials (AEP), reaction times and mood scales. RESULTS: In all subjects the pain related variables were suppressed maximally 2 h after morphine administration (p < 0.01 versus placebo), indicated by a decrease of the pain ratings by about 45% and of the pain related brain potentials by about 50%. Interestingly, no effect on any sedation variable was found (p > 0.05). CONCLUSION: The lack of sedative effects in the presence of marked analgesia was surprising in comparison with results of previous studies. It is concluded that the experimental pain increased the arousal level thus counteracting morphine-induced sedation. This may explain why other studies found relevant sedation after morphine application in the absence of pain. This underlines that sedative effects of analgesic drugs should be evaluated in the presence of pain. In relation to other analgesics (meperidine, pentacozine, nortilidine, flupirtine and tramadol) evaluated by exactly the same experimental protocol, morphine exhibited a potent analgesia with the smallest sedative effects of all. PMID- 15112861 TI - Influence of losigamone on the pharmacokinetics of a combined oral contraceptive in healthy female volunteers. AB - OBJECTIVES: The influence of the new antiepileptic drug losigamone (CAS 112856-44 7/123783-52-8) on the pharmacokinetics of a combined oral contraceptive containing ethinylestradiol (CAS 57-63-6) and levonorgestrel (CAS 797-63-7) was investigated in 16 healthy women. METHODS: This phase I study consisted of 3 periods with an uncontrolled first period and a double-blind, placebo-controlled, cross-over design in the second and third period. All subjects received a single dose of 200 mg losigamone (1 tablet) in period 1 (on day 14) as well as multiple doses of losigamone (3 tablets = 600 mg per day) or placebo for 15 days in periods 2 and 3. During all three periods an oral contraceptive containing 30 microg ethinylestradiol and 150 microg levonorgestrel was given. Single-dose pharmacokinetics was investigated on day 14 of period 1. Multiple-dose pharmacokinetic investigations were performed on day 15 of periods 2 and 3. The samples were assayed to derive pharmacokinetic data of ethinylestradiol and levonorgestrel. In addition, the concentrations of losigamone racemate (AO-33) and its enantiomers AO-242 and AO-294 were determined in these samples. RESULTS: The mean values of the pharmacokinetic parameters AUC and Cmax of ethinylestradiol and levonorgestrel after multiple-dose treatment with losigamone or placebo were quite similar and met the criteria for bioequivalence. The 90% confidence intervals of the log-transformed ratios of the geometric means of the primary pharmacokinetic variables were included in the respective acceptance ranges of 80% to 125% (AUC) and 70% to 143% (Cmax). CONCLUSIONS: The study demonstrated that multiple doses of losigamone did not influence the multiple dose kinetics of ethinylestradiol and levonorgestrel. The single- and multiple dose kinetics of 200 mg losigamone and its enantiomeres did not differ from each other in a significant way. The combination of losigamone and the combined oral contraceptive was well tolerated and no serious adverse events occurred. It can be stated that the antiepileptic drug losigamone and the combined contraceptive do not interact each others metabolism. PMID- 15112862 TI - Evaluation of the bioequivalence of two tablet formulations of enalapril/hydrochlorothiazide after single oral administration to healthy volunteers. AB - The pharmacokinetic parameters of two oral formulations of 20/12.5 mg tablets of enalapril/hydrochlorothiazide (CAS 75847-73-3 and CAS 58-93-5, respectively; Penopril as test and another commercially available preparation as reference) were compared in an open-label randomized single oral dose two-period cross-over design to 24 healthy volunteers under fasting conditions. Plasma concentrations of enalaprilat (CAS 76420-72-9), the pharmacologically active metabolite of enalapril, and hydrochlorothiazide were determined by a validated GC/MS and HPLC assay, respectively. Serial blood samples were collected prior to each administration and at 19 timepoints within 36 h after dosing. The parametric 90% confidence intervals of the geometric mean values of the test/reference ratios for enalaprilat were 99.3% to 118.9% (point estimate: 108.7%) for AUC(0 infinity), 97.3% to 116.9% (point estimate: 106.7%) for AUC(0-t), and 92.5% to 113.0% (point estimate: 102.3%) for Cmax, and for hydrochlorothiazide 92.3% to 105.1% (point estimate: 98.5%) for AUC(0-infinity), 92.7% to 105.4% (point estimate: 98.9%) for AUC(0-t), and 97.6% to 115.3% (point estimate: 106.0%) for Cmax, within the acceptance criteria for bioequivalence (80%-125%). Tmax values were analyzed by the nonparametric Wilcoxon test and the difference was not statistically significant. Therefore, it is concluded that the test and reference enalapril/hydrochlorothiazide formulations are bioequivalent for both the extent and the rate of absorption. PMID- 15112863 TI - Pharmacokinetics and bioequivalence study of a generic desloratadine tablet formulation in healthy male volunteers. AB - The pharmacokinetic profiles and relative bioavailability of desloratadine (CAS 100643-71-8, Denosin as test and another commercially available preparation as reference) tablets from two different pharmaceutical manufacturers were carried out. A single oral dose (10 mg/2 tablets) of desloratadine was administered to 8 healthy young Chinese males in a completely double-blind cross-over design with a two-week washout period between each dose. Plasma samples were obtained before and at various appropriate intervals after dosing up to 120 h. The plasma concentrations were then analyzed by a liquid chromatography/tandem mass spectrometric (LC/MS/MS) method. The limit of quantitation of this LC/MS/MS method was 0.05 ng/mL. The coefficients of variation of the within-day and between-day calibration curves (n = 6) range from 0.05 ng/mL to 10 ng/mL and were less than 10%. The accuracy of this method was verified. Values for the area under the plasma concentration-time curve (AUC), peak concentration (Cmax), time to peak concentration (Tmax), elimination rate constant, half-life, oral clearance were estimated and compared for each preparation. By ANOVA, 90% confidence interval, Mann-Whitney test, and paired t-test, the two desloratadine products can be considered bioequivalent for both the extent and the rate of absorption. PMID- 15112864 TI - Absorption, distribution and excretion of 14C-pilocarpine following oral administration to rats. AB - The absorption, distribution and excretion of pilocarpine (CAS 92-13-7) were studied after single oral doses of 14C-pilocarpine hydrochloride (CAS 54-71-7) to the Sprague-Dawley rat, administered in aqueous solution mainly at a dose level of 0.3 mg/kg. Rats also received single intravenous doses at 0.3 mg/kg so as to compare 14C pharmacokinetics and excretion. The oral 14C-dose was rapidly and almost completely absorbed from the duodenum and small intestine within 30 min in the male rat and 14C concentrations in plasma declined biexponentially with a terminal half-life of about 9 h. Over the oral dosage range studied, i.e. 0.1-1.0 mg/kg, there was no evidence of significant non-proportionality for Cmax of 14C, whereas there was some such evidence for AUG24. Tissue 14C concentrations in male and pregnant female (Day 18) rats peaked at 0.5 h and mostly declined in parallel with those in the plasma. Excluding tissues concerned with drug absorption and elimination, 14C concentrations in most tissues were similar to, or lower than, those in the plasma. The extent of placental transfer of 14C was small and less than 0.09% of a maternal dose reached a foetus. 14C diffused into maternal milk at concentrations similar to those in the plasma. The 14C-dose was rapidly excreted in male rats, mostly in the urine (about 80%) during 6 h post dose. Recoveries of 14C in mass balance (excretion) studies were in the range 96-100%. There were no apparent gender differences in the disposition of 14C-pilocarpine in the rat. PMID- 15112865 TI - Fertility study of the new pyrazolopyrimidinone derivative DA-8159 for erectile dysfunction in rats. AB - DA-8159 (5-[2-propyloxy-5-(1-methyl-2-pyrrolidinylethylamidosulfonyl)phenyl]-1 methyl-3-propyl-1,6-dihydro-7H-pyrazolo(4,3-d)pyrimidine-7-one, CAS 268203-93-6) is a new pyrazolopyrimidinone derivative for the treatment of erectile dysfunction. The test agent was administered by gavage at dose levels of 0, 17.5, 70, and 280 mg/kg to Sprague-Dawley male rats from 28 days before mating to the end of the mating period, and to females from 14 days before mating to day 6 of gestation. Effects of the test agent on general findings and reproductive performance of parent animals and on early embryonic development were examined. At 280 mg/kg, salivation and hair loss were observed at a high incidence in males and females, respectively. One female out of 24 died during the treatment period. A decrease in food consumption and a decrease in body weight were observed in both sexes. No treatment-related gross findings and histopathological findings were seen, except that the dead animal exhibited atrophy of the thymus. A decrease of prostate weight and an increase of liver, lung and spleen weights were seen. An increase in the period and irregularity of sexual cycle was observed. A decrease in fertility and pregnancy index was also seen. A decrease in the number of corpora lutea, implantations, and litter size was observed. There were no treatment-related changes in precoital time, mating index, sperm parameters, and serum testosterone concentration. At 17.5 mg/kg and 70 mg/kg, there were no adverse effects on all the parameters examined. Based on these results, DA-8159 induces suppressed body weight, decreased food consumption, irregularity in sexual cycle, reduced fertility, and decreased number of corpora lutea, implantations, and litter size at 280 mg/kg and no observed adverse effect levels (NOAELs) of DA-8159 are considered to be 70 mg/kg for general toxicity and reproductive capability of parent animals and for early embryonic development, respectively. PMID- 15112866 TI - Effect of the new H1-antagonist ReN1869 on capsaicin-induced hyperalgesia in human skin/Human phase-I trial using somatosensory evoked potentials induced by a CO2 laser. AB - Extensive pre-clinical investigations have shown that the tricyclic compound ReN1869 ((R)-1-(3-(10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-ylidene)-1-propyl) 3-piperidine carboxylic acid, CAS 170149-99-2) is a potent H1-antagonist with pronounced antinociceptive properties. In this human phase-I trial the effect of different acute and multiple doses of ReN1869 on capsaicin induced neurogenic inflammation and hyperalgesia was investigated. Twenty-one healthy male subjects were enrolled in this randomised, double-blind, three-period, crossover trial design--consisting of acute and one week b.i.d. oral administration of 25 and 50 mg doses of ReN1869 and matching placebo--separated by 3 week washout periods. Capsaicin solution (1%) (INCI: Capsicum frutescens--containing capsaicinoides from Capsicum annuum annuum, CAS 84603-55-4) was applied in an occlusive mode for 30 min on the skin of the back in all three acute and subchronic medication periods to induce neurogenic inflammation. When the nociceptive laser pulses were applied to the capsaicin pre-treated skin, ReN1869 exerted a highly significant reduction of the pain response--as predominantly detected by suppression of the (central) P2-component in the laser-induced somatosensory evoked potentials (LSEPs) from Vertex-EEG. The primary efficacy endpoint, the N1/P2 peak to peak amplitude, was significantly reduced with the administration of ReN1869- primarily by a suppression of the P2-component of the LSEP. This suppression was dose-dependent and was more pronounced after a one week treatment (subchronic mode) with ReN1869 than after the first dose (acute mode). In contrast to the (central) P2-component there was no significant effect on the (peripheral) N1 component of the LSEPs taken from capsaicin-treated skin. As ReN1869 had no significant effect when the laser pulses were applied to normal skin, and the compound's effect was mainly restricted to the (central) P2-component, when LSEPs were taken from capsaicin treated skin, it can be concluded that ReN1869 exerts its positive effect to reduce capsaicin-induced hyperalgesia by a primarily central mechanism. PMID- 15112867 TI - [Brachytherapy: notable or not able]. PMID- 15112868 TI - [Management of staghorn calculi]. AB - OBJECTIVES: More than 200 articles about treatment of staghorn calculi have been published over the last 15 years; we observe a progressive tendency to the elective indication of a combination of therapeutic methods. Recent development of flexible ureteroscopy and endoscopical holmium-YAG laser lithotripsy has prompted the application of a new method of combined treatment for staghorn calculi: retrograde ureteroscopy and external shock wave lithotripsy. The objective of this work is to analyze indications and results of various therapeutic methods in relation to staghorn calculi complexity. METHODS AND RESULTS: We analyze the results of open surgery, extracorporeal shock wave lithotripsy, percutaneous nephrostomy and retrograde ureteroscopy in the treatment of staghorn calculi Group 1 (soft calculi < 700 mm2 of area and homogeneous pyelocalyceal distribution), Group 2 (calculi < 700 mm2 hard or with predominantly central pyelocalyceal distribution, wide infundibula and calyces with few branches), Group 3 (calculi > 700 mm2 with predominantly central or homogeneous distribution, absence of infundibular stenosis or excluded calyces) and Group 4 (great lithiasic mass predominantly homogeneous or peripheral, narrow infundibula or multiple infundibular stenosis and/or lithiasis within excluded calyces). CONCLUSIONS: In Group 1 staghorn calculi ESWL (JJ catheter + ESWL) has good results in 62.5% - 72.5% of the patients after an average of 3.2-3.6 sessions. The combination of contact lithotripsy by retrograde ureteroscopy and ESWL may be a therapeutic alternative. In group 2, percutaneous renal surgery may be considered the technique of choice; persisting residual fragments are treated by ESWL. Group 3 calculi are indication for combined therapy, percutaneous renal surgery and ESWL. ESWL therapy only is not indicated; open surgery through a sinus approach combined with intraoperative pyelocalyceal nephroscopy may be considered an option. Open surgery is recommended for Group 4 calculi--sinus approach is preferred--with mechanical extraction of caliceal fragments and/or pyelocalyceal nephroscopy support. PMID- 15112869 TI - [Improvement of anemia and quality of life of patients with prostate cancer by treatment with human recombinant erythropoietin]. AB - OBJECTIVES: Anemia is a frequent finding in patients with prostate cancer. Reduction of erythropoiesis caused by androgenic blockade is among its etiologies. Therefore, quality of life of these patients results decreased, being origin of significant morbidity and mortality. Recombinant forms of human erythropoietin have demonstrated their effectiveness improving quality of life of patients with various solid tumors, but specific studies in prostate cancer are a few. Our objective is to evaluate the efficacy of human recombinant erythropoietin (EPO) correcting anemia and improving the quality of life of patients with prostate cancer. METHODS: Prospective study of patients with prostate cancer under combined androgenic blockade treatment having hemoglobin levels lower than 11 g/dl. We analyze clinical characteristics and quality of life prior to EPO dispense. We used the Triple Linear Analogical Scale for patients with cancer (CLAS) to evaluate quality of life. EPO was administered during 12 weeks (10.000 units subcutaneously, three times a week). We study the evolution of different parameters compared to baseline. RESULTS: We included a total of 17 patients. Average age was 75.5 +/- 5.9 yr. Average time of neoplasia evolution was 57.6 +/- 13.9 months. Eleven patients presented active disease (PSA > 1 ng/ml). Mean initial haemoglobin level was 10.3 +/- 0.4 mg/dl, reaching 12.2 +/ - 1.0 after treatment (p < 0.001). Patients with active disease and levels of initial haemoglobin smaller than 10.2 g/dl presented worse outcomes. There were not adverse events attributable to EPO. CONCLUSIONS: We consider that the administration of EPO increases significantly the levels of haemoglobin and the quality of life of patients with prostate adenocarcinoma, being the response worse in patients with low levels of baseline haemoglobin. PMID- 15112870 TI - [Treatment of urethral stenosis by self expandable endourethral prosthesis. Long term results]. AB - OBJECTIVES: Urethral stenosis is a frequent disease in males. The variable results of its treatment have prompted the surgeon to search for new therapeutic alternatives. We propose to treat it by self expandable endourethral prosthesis. METHODS: Since February 1993 to March 2000 48 patients with urethral stenosis previously treated with endoscopic urethrotomy have undergone the procedure. RESULTS: All patients completed a minimum of 3 years of follow-up. Patients lost to follow-up for any cause were not included in the final study description. All patients had satisfactory micturition at the time of discharge. Mean flow on uroflowmetry at three years was higher than 14 ml/ s in 87.5% (42/48 patients evaluated). CONCLUSIONS: The use of self expandable endourethral prosthesis is a valid alternative in the treatment of recurrent urethral strictures due to its good long-term results and low morbidity. We recommend not using it in cases of traumatic etiology. PMID- 15112871 TI - [Antibiotics in transurethral resection of the prostate in patients with low risk of infectious complications: randomized prospective comparative study]. AB - OBJECTIVES: To compare the rate of infectious complications using 2 antibiotic schemes in prostatic transurethral resection of the prostate (TUR-P) of patients at low risk, in order to reduce the use of antibiotics in this kind of patients. Secondarily, try to weigh the influence of clinical background, intraoperative complications and postoperative outcome on the development of such complications. METHODS: A comparative, prospective, randomized, open study was designed including 95 patients with sterile urine without indwelling catheter, subjected to TUR-P during one year. Group 1 received cefazolin 1 gr. i.v. preoperative and every 8 hrs. during the first day (3 doses) followed by ciprofloxacin 250 mg. oral every 12 hrs until the catheter was removed (therapeutic dose). Group 2 received cefazolin 1 gr. i.v. preoperative and at 8 hrs postoperative (2 doses) followed by nitrofurantoin 100 mg. oral every night until the catheter was removed (prophylactic dose). Five patients were excluded after randomization (5.3%) and all the remainders completed follow up. RESULTS: Ninety patients are analyzed, 45 in each group. Both groups were well matched with regard to clinical background, surgical and postoperative parameters and complications. Fever (axillary temperature equal or over 37.5 degrees C) was present in 2% of Group 1 and 11% of Group 2 (p = 0.091). Postoperative early or late bacteriuria (colony count > 100,000 CFU/mL) was present in 2% of Group 1 and in 13% of Group 2 (p = 0.049). Postoperative urinary infection (bacteriuria + clinical infection) was present in 2% of Group 1 and in 16% of Group 2 (p = 0.026). A statistical association was found between fever and postoperative urinary infection in all patients (p = 0.029) and between purulent secretion during prostatic tissue cutting and fever in Group 2 (p = 0.01). CONCLUSION: Patients in Group 1 (cefazolin-ciprofloxacin) presented significant less postoperative urinary infection than those in Group 2 (cefazolin-nitrofurantoin) represented by less postoperative bacteriuria frequency. This was possibly due to different antimicrobial activity and dosage of used drugs. Fever was statistically related to postoperative urinary infection. PMID- 15112872 TI - [Our experience with simplified vasovasostomy. Review of our results during the last 5 years]. AB - OBJECTIVES: We currently see an increase of demand of vasectomy reversion, mainly due to the wish of recovering fertility. METHODS: We review 21 cases of vasovasostomy performed at our department over the last 5 years with the single layer technique under optical magnification, comparing our results with those from bibliography for both vasovasostomy and in vitro fecundation techniques used in patients with obstructive azoospermia, specifically intracytoplasmic spermatozoids injection (ICSI). RESULTS: Success rates obtained for permeability, pregnancy, and births were 80%, 33%, and 26.66% respectively. CONCLUSIONS: Our results in terms of pregnancies are equivalent to those obtained by ICSI. These data, together with the disadvantage of greater rates of multiple pregnancies associated with in vitro fecundation techniques, moved us to choose vasovasostomy as the initial option for treatment of male infertility secondary to vasectomy. PMID- 15112873 TI - [Infiltrating bladder carcinoma and adrenal incidentaloma. Report of one case. Bibliographic review]. AB - OBJECTIVES: We report one case of left adrenal tumor discovered during the study of an infiltrating bladder carcinoma. We discuss the diagnostic and therapeutic implications of the clinical case. METHODS: Surgical procedure was undertaken with radical cystectomy and Bricker's type urinary diversion, plus adenomectomy by anterior approach. RESULTS: The pathologic report showed a myelolipoma. CONCLUSIONS: Most adrenal tumors are benign, mainly if they are small and non functioning. The protocol of hormonal study should include potassium and catecholamines. Tumors greater than 6 cm and functioning masses are indications for surgery. For oncological patients, surgical approach of the lesion is indicated if there is not evidence of dissemination. PMID- 15112874 TI - [Bladder mucormycosis. Report of one case]. AB - OBJECTIVES: To report one case of an opportunistic bladder infection by fungus of the Mucorales order. METHODS: An old patient with permanent bladder catheter due to cerebral vascular accident presented with hematuria associated with change in his general health status, and a bladder mass on ultrasound Cystoscopy and cold biopsy were performed RESULTS: Pathology reported bladder mucormycosis. CONCLUSIONS: Mucormycosis is an opportunistic infection by fungus of the Mucorales order, which disseminate hematogenously from an entrance. Diagnosis is usually histological, seeing characteristic hyphae and ischemic or hemorrhagic necrosis. Bladder involvement is very uncommon. Treatment is based on the combination of debridement of the necrotic areas and intravenous antifungal drugs (amphotericin B). PMID- 15112875 TI - [Continent urinary diversion after complete urethral resection: the Mitrofanoff technique]. AB - OBJECTIVE: To describe our experience using the Mitrofanoff procedure in adult patients after a complete urethral resection. METHODS AND RESULTS: We describe 2 clinical cases, both with neoplastic lesions in the urethra. Once completed the urethral resection, we made a continent urinary diversion using the Mitrofanoff procedure with the cecal appendix, applying a few technical variations. CONCLUSIONS: The Mitrofanoff procedure is a simple technique, applicable to both pediatric and adult patients and it should be considered a good urinary diversion after complete urethral resection. PMID- 15112876 TI - [Lithiasis of soft appearance within a calyceal diverticulum in the right kidney. Report of one case and therapeutic management]. AB - OBJECTIVES: To report the case of a 28-year-old female patient carrying a lithiasis of soft appearance located within a calyceal diverticulum in the right kidney which presented clinically during pregnancy, and to comment on therapeutic management, as well as the outcome one year after conservative treatment. METHODS AND RESULTS: 28-year-old female patient with history of renal colic during pregnancy, who presents at our clinics in the postpartum period with a recurrent abdominal and lumbar pain. Kidney-ureter-bladder (KUB) x-ray and intravenous urography (IVP) detected a lithiasis of soft appearance within a calyceal diverticulum in the right kidney. Treatment was conservative with potassium citrate/citric acid and follow-up controls with urine culture (every 3 months) and ultrasounds (every 8 months). Currently she is asymptomatic but the lithiasis persists on KUB and urine culture is positive (Escherichia coli) and receives treatment with cefuroxime 250 mg every 12 hours for 6 days and nitrofurantoin 100 mg every night for three months. CONCLUSIONS: Most urinary stones located within calyceal diverticula have an asymptomatic course, treatment not being necessary. The most frequent clinical presentations of these stones are flank pain, urinary infection demonstrated by positive urine culture, and incidental finding. IVU is an effective method for diagnosis. ESWL is not an effective treatment for them, being percutaneous techniques the most suitable for a single-surgical procedure resolution of both lithiasis and pyelocalyceal diverticulum. PMID- 15112877 TI - [Migration of an intrauterine contraceptive device into the urinary bladder: report of one case]. AB - OBJECTIVES: To report one case of uterine perforation and migration into the urinary bladder of an intrauterine contraceptive device. METHODS/RESULTS: 42-year old female patient who presents with lower urinary tract irritative syndrome in association to recurrent urinary tract infection. Ultrasound revealed apart of an intrauterine device inside the bladder, device which was inserted years before and was supposed to have come out spontaneously. Urethrocystoscopy with extraction of the intravesical segment and hysteroscopy with extraction of the intrauterine segment were carried out. CONCLUSIONS: The postoperative period was satisfactory and patient is currently asymptomatic. Radiological or ultrasound controls should be performed in the follow-up of patients with intrauterine contraceptive devices. The inability to locate an intrauterine contraceptive device in a patient who did not realize it coming out should be considered an uterine perforation until proved otherwise. PMID- 15112878 TI - [Large cell calcifying Sertoli cell tumor of the testis: report of one case]. AB - OBJECTIVES: To report one case of Sertoli cell testicular tumor of the large cell calcifying type, a rare presentation, and to perform a medical literature review on the topic. METHODS: Our case report is based on the patient's medical history and images of the diagnostic tests and pathology. We perform a bibliographic review of the available literature in MEDLINE. RESULTS: The case is reported with the diagnostic tests and pathology report. We present a comprehensive review of the medical literature on the topic. CONCLUSIONS: The large cell calcifying Sertoli cell tumor is a rare testicular tumor of benign behaviour. Due to its benign behaviour, treatment of this kind of tumor should be conservative with resection of the lesion, preserving the testicle. PMID- 15112879 TI - [Metachronous recurrence of idiopathic high flow priapism]. AB - OBJECTIVES: To report one case of metachronous recurrence of idiopathic high flow priapism. METHODS. We describe the case of a 28-year-old male patient who presents with penile partial tumescence which started 10 hours before and history of a similar episode seven years before. Physical examination confirmed the clinical picture, with mild local discomfort on palpation. Blood tests were normal. Cavernous blood gases were compatible with arterial blood O2 saturation levels. The patient had a satisfactory progressive response to oral administration of diazepam. No pathologic findings were seen at the time of arteriography. RESULTS: Complete resolution of the episode. Erectile function was satisfactorily preserved. There was no relapse after one year of follow-up. CONCLUSIONS: We remark the validity of a deferred therapeutic attitude in front of pictures of high flow priapism, as well as the possibility of resolution with conservative measures. We emphasize the peculiarities of this case, mainly its idiopathic character and the metachronous recurrence. PMID- 15112880 TI - Transforming growth factor-beta2 and beta3 expression in carcinoma of the prostate. AB - OBJECTIVES: The present study reports the clinical and histological features of 14 cases of prostate adenocarcinoma coupled with their expression of the TGF isoforms beta2 and beta3, as well as their receptor endogline (CD105). METHODS: Fourteen (14) cases of adenocarcinoma (ADC) of the prostate were examined. Relevant clinical data were gathered From the histological point of view, the tumor's grade and the evidence of perineural, vascular and/or lymphatic invasion were the key elements taken into account. Immunohistochemical analyses were carried out to assess the expression of TGFbeta-2, TGFbeta-3 and CD105 in tumoral tissue samples. RESULTS: Patient's age ranged between 57 and 74 years. Thirteen (13) had prostatic specific antigen (PSA) values above the 4 ng/dL threshold Eleven (11) ADCs were moderately differentiated. The predominant grade in relation to the nucleus was II/III (7 cases). In two (2) of the 14 cases, no grading classification was applicable as the tumors exhibited changes related to the effect of hormonal therapy. In eleven cases, expression of TGFbeta-2 was detected in the cytoplasm of tumoral cells. Two cases also showed focal expression of TGFbeta-3, as well as in prostatic intraepithelial neoplasia areas of a third case. The areas with the highest intensity of expression were those occupied by basal cells and regions of glandular atrophy. These were also the areas of CD105 expression. CONCLUSIONS: Our results allow us to conclude that the TGF-beta3 family of cytokines, particularly the isoforms beta-2 and beta-3, seem to play a key role in the initiation, progression and transformation of tumoral cells inprostatic ADC. Future studies should be directed to the full understanding of the impact that these factors exert on tumoral behaviour, with an emphasis on those steps susceptible to therapeutic manipulation. PMID- 15112881 TI - The rediscovery of continuous flow peritoneal dialysis. PMID- 15112882 TI - Ozone therapy. PMID- 15112883 TI - Extracorporeal support of liver function (II part). PMID- 15112884 TI - Artificial in vivo biofiltration: slow continuous intravenous plasmafiltration (SCIP) and artificial organ support. AB - An intravenous plasmafiltration (SCIP) catheter has been developed and is proposed for clinical investigation into the alleviation of acute fluid overload by SCUF of the extracted plasma. The system utilizes a unique backflushing technique, high intravenous shear flow rates and biocompatible polymers to minimize protein and platelet aggregation along the filter surfaces. The absence of platelets from the extracted plasma promotes the longevity of ultrafiltration cartridges, thus theoretically minimizing attendant labor associated with continuous renal replacement therapies. Clinical studies are currently being planned for the near future. Plasma SCUF is envisioned as a predecessor technology to future applications in therapeutic apheresis, tissue engineering, therapeutic sorbent technologies. Further, with improved longevity profiles, intravenous SCUF or dialysis and implantable or wearable artificial organs based upon artificial in vivo biofiltration are possible. PMID- 15112885 TI - Convective-controlled double high flux hemodiafiltration: a novel blood purification modality. AB - Convective-controlled double high flux hemodiafiltration (CC-DHF) was set-up using two high flux dialyzers. The convection occurred in the first while the fluid replacement took place in the second dialyzer. The system of CC-DHF basically resembled that of hemodiafiltration. CC-DHF was performed in 9 chronic hemodialysis Thai patients who had been treated with high flux hemodialysis for at least 6 months. When compared with high flux hemodialysis, CC-DHF could provide higher Kt/Vurea (2.4+/-0.4 vs. 2.0+/-0.4, p<0.05) and beta2-microglobulin clearance (106.2+/-15.4 vs. 48.9+/-6.1 ml/min, p<0.01). Following 6-month therapy of CC-HDF, the predialysis beta2-microglobulin levels were reduced by 12.7% while the values of Kt/Vurea were consistently higher than 2.7. The quality of life consistently improved during the 6 months of CC-DHF treatment. There were no differences in clinical and technical complications between CC-DHF and high flux hemodialysis. In conclusion, CC-DHF could provide performance comparable to hemodiafiltration without the need for expensive hemodiafiltration machines. PMID- 15112886 TI - Diffusive clearance of small and middle-sized molecules in combined dialyzer flow configurations. AB - Clearance of low (LMW) and middle molecular weight (MMW) solutes was investigated in vitro for different dialyzer configurations and mutual flow directions. Single pass tests were performed with two low flux Fresenius F6HPS hemodialyzers placed in series (12 tests) and in parallel (6 tests), and results were compared with those for one single dialyzer (2 tests). Either high concentrated (45mS/cm) bicarbonate dialysis fluid (surrogate LMW) or trisodiumphosphate (surrogate MMW) concentration (31mS/cm) was used as blood substitution fluid. Standard blood and dialysate flows of 250 and 500ml/min, respectively, were prescribed. Clearance was derived from conductivity measurements in blood and dialysate compartment, correcting for the overall ultrafiltration rate of 0.1-0.5l/h. In a single dialyzer, changing the counter current flow to co-current deteriorates diffusive clearance by 14% (LMW) and 18% (MMW). Compared to one single dialyzer using counter current flow, clearance increases by 3 to 8% (LMW) and by 15 to 18% (MMW) using two dialyzers in parallel and in series, respectively. As a consequence, the benefit by using a second dialyzer is more prominent for larger molecules. Moreover, pressure profiles drawn for the different configurations show the impact of limited convection on diffusive clearance. PMID- 15112887 TI - Von Willebrand factor and autoantibodies against oxidized LDL in hemodialysis patients treated with vitamin E-modified dialyzers. AB - Oxidant stress is a well known cause of damage in the atherosclerotic process. Vitamin E is one of the most promising natural antioxidants. In this study we investigated if a vitamin E-coated dialyzer was able to reduce the plasma levels of auto-antibodies against oxidized-LDL, von Willebrand factor (vWf) and thrombomodulin (TM) as markers of endothelial damage. In this controlled 6-month prospective study, we investigated these markers in two matched groups (n=16 each) of patients on regular hemodialysis not yet diagnosed for atherosclerosis cardiovascular disease (ACVD) (mean age=58.3+/-7.0 yrs, mean dialysis age=30.1+/ 10.0 months), in which cellulosic (CLS) and vitamin E-modified dialyzers (CLE) were compared. At inclusion all the patients were treated with CLS. Then, the study group was shifted to CLE for 6 months. At baseline the patients showed normal levels of vitamin E and high levels of oxLDL-Ab, vWf and TM compared to healthy subjects. In the CLE group oxLDL-Ab and vWf, but not TM levels, decreased progressively (from 472+/-287 to 264+/-199 mU/mL, p<0.0001 and from 101.1+/-7.5% to 76.7+/-18.5%; p<0.001, respectively), and vitamin E increased from 4.40+/-0.81 to 7.81+/-1.16 microg/mg of cholesterol. At the end of the study, 8 of the patients treated with CLE were randomly selected and went back to the membrane without Vitamin E for six months. They showed an significant increase in OxLDL-Ab and vWf levels and a significant reduction in tocoferol levels. In conclusion, CLE compared to cellulosic dialyzers can lower some indices of damage to LDL and endothelial cells. PMID- 15112888 TI - Numerical simulation of the hepatic circulation. AB - Availability of donor livers and the relatively short preservation time limit the success of liver transplantation. The use of hypothermic machine perfusion could pave the way for expansion of the donor pool. To better define optimal settings of such a device, the feasibility of using a numerical simulation model of the hepatic circulation is determined. Hemodynamics in the hepatic arterial, portal venous and hepatic venous compartments of the hepatic vascular tree was modelled using an electrical analogue. Calculated pressure and flow profiles throughout the liver were in accordance with physiologic profiles in the total circulatory system. Comparison of calculated flow values with normal control values showed a discrepancy that was explained by inaccurate diameter input data. Until more precise methods for determining vascular dimensions become available, redefining vessel diameter makes the simulation model perfectly suitable for predicting influences of temperature and/or viscosity on hepatic hemodynamics and is thereby an excellent tool in defining optimal settings for our hypothermic liver perfusion system. PMID- 15112889 TI - Proposal for a quantitative description of blood spiral flow in medical devices. AB - The association between specific blood flow patterns and blood behaviour through medical devices suggests that a Lagrangian study may be a useful instrument for the evaluation of the thrombogenic and/or hemolytic potential of certain devices' geometries and biomaterials. In this study a description of blood particle trajectories in terms of their spiral contents is proposed; such a mathematical description for blood spiral flow, computed along several pathlines, is tested for a quantitative determination of the spiralled motion of blood flow into two three-dimensional numerical models, having different design characteristics, of venous cannula inserted in a vessel. As the influence of vortical flow conditions have been observed to have both beneficial and detrimental influence on blood behaviour in terms of blood-device interaction, of the degradation of its components, and of the efficiency of mass-exchange (in red cells oxygenation and plasma filtration, for example), the herein proposed method for the description of spiral laminar motion may be a helpful instrument to build up a tool to investigate, for example, the existence of correlations between level of spiral flow and geometry (as in the present investigated test case), rather than the effects of blood-surface contact. The results obtained in this test case investigation, confirm the effectiveness of the proposed function for a quantitative analysis of spiral flow in medical devices. PMID- 15112890 TI - Effect of left ventricular assist device on circulatory autonomic nervous activity. AB - PURPOSE: For evaluation of the autonomic nervous system (ANS) activity of patients with mechanical circulatory support including LVAD, IABP and PCPS, arterial pressure variability (APV) was examined. APV is the power density values of pressure signals. High frequency (0.15 to 0.4 Hz) and low frequency (0.04 to 0.15 Hz) components of APV are regarded as a function of the autonomic nervous system. METHODS: Arterial pressure signals were acquired every 2 hours at 100 Hz for 6 minutes. Power spectral analysis was carried out, and APV was observed in 18 patients with IABP and/or PCPS. APV was also observed in 3 patients with LVADs (Xemex, Zeon Medical, Japan) before their discharge from ICU for transfer to another hospital for cardiac transplantation. RESULTS: Although the value gradually increased in 18 patients with IABP and/or PCPS, sustained low values of LFAnorm of APV in 6 patients were related to prolonged mechanical support, and 4 patients among them died consequently. The normalized power spectral component with low frequency (LFAnorm) was maintained at relatively high values in 3 patients with LVADs. CONCLUSION: The study results suggest that power spectral analysis of systemic arterial pressure offers a reasonable means for the evaluation of the severity of patient cardiovascular condition and for the prediction of clinical outcome. It also suggested that LVAD support exerts a favorable effect on ANS in patients with severe heart failure before cardiac transplantation. PMID- 15112891 TI - Maintenance of residual renal function 10 years after the start of hemodialysis: the advantage of tailored schedules? AB - Maintenance of residual renal clearance is a clinical advantage, protecting against the long-term effects of uremia: although demonstrated in peritoneal dialysis, the strategies in hemodialysis are less clear. This case suggests that dialysis schedules individualized on the basis of renal clearances may help preserve residual function. SB is a 58 year-old male who started dialysis in emergency (creatinine 30.7 mg/dL) in 1993. He had a history of gout, small shrunken kidneys and moderate hypertension. The clinical diagnosis was vasculointerstitial nephropathy. Eighteen months after starting hemodialysis on a conventional thrice weekly schedule, the patient was switched to 2 sessions/week (creatinine clearance increased to 6 ml/min). Thereafter, clearances were checked in alternate months and treatment was tailored to an equivalent renal clearance > or =12 ml/min (1-2 sessions, 2-3.30 hours/week). Ten years after beginning dialysis, he is on a twice weekly schedule (3.30 hours), is normotensive, works full-time and does not want to go on a transplant waiting list. PMID- 15112892 TI - [Image of the month. Atypical mass of the right atrium]. PMID- 15112893 TI - [Image of the month. Merkel cell tumor: response to chemotherapy]. PMID- 15112894 TI - [Clinical case of the month. Autoimmune cirrhosis treated by liver transplantation using the right hepatic lobe from a living related donor]. AB - The authors describe the case of a 17-year-old girl who suffered from end-stage liver failure due to chronic autoimmune hepatitis. Liver failure was complicated by severe portal hypertension, hypersplenism and refractory ascites. Liver transplantation was indicated. She was listed for cadaveric whole liver transplantation, but her infrequent blood group (B) increased waiting time. Her condition deteriorated to Child C liver failure and living related liver transplant was considered. Her father was compatible and proposed himself for donation. Right lobe procurement was decided in order to provide sufficient liver mass. No transfusion of red cells, platelets, or fresh frozen plasma was used either in the donor or the recipient. Both recipient and donor left the ward at postoperative day 14, without complication. They were both asymptomatic and with normal liver tests at one year follow-up. Living related liver transplantation using the right lobe may offer an alternative to liver transplant candidates in this period of organ donor shortage. PMID- 15112895 TI - [The top ten advances in heart disease and stroke research for 2003: a selection from the American Heart Association]. AB - Since 1996, the American Heart Association (AHA) publishes, with its year-end report, the list of "The Ten Top Advances in Heart Disease and Stroke Research". These milestones include the results of some major clinical trials, new medications, guidelines published during the past year, and innovative research work that can be very preliminary, but contains promising data for the years to come. It is not without interest to read the list of publications or presentations that were selected; one can indeed imagine that these contributions were, over the past twelve months, among the most significant in our specialty and that some of them will indeed exert a considerable influence on the practice of cardiology in the future. It is with pleasure that we observe that some of these publications were already analysed in our journal during 2003. PMID- 15112896 TI - [Chronic pain management: a multidisciplinary approach]. AB - Assessment of chronic pain is one of medicine's most difficult challenges. A structured and flexible multidisciplinary approach allows full characterisation of the various components of the pain syndrome. This then allows the use of a rational combination of pharmacologic, physical, psychological, and surgical techniques. It is essential to gain the patient's confidence, collaboration, and compliance. Patients can better manage their situation when their needs are clarified and when care is oriented toward concrete therapeutic objectives. PMID- 15112897 TI - [Primary mycotic aneurysm of the infrarenal aorta: a report of 2 cases]. AB - Infrarenal mycotic aneurysms are rare. The classic symptomatic triad is abdominal pain, fever and a pulsatile, rapidly growing abdominal mass. We present two cases of mycotic abdominal aneurysm, recently managed. The principles of diagnosis and treatment of mycotic aneurysm are reviewed and discussed. PMID- 15112898 TI - [Multiple myeloma and HIV infection: report of 3 cases]. AB - HIV infection rages at the endemic state in Sub Saharan African and especially in Congo Brazzaville. We report the observation of three female patients infected with HIV and developing multiple myeloma. The three patients were treated at the University hospital of Brazzaville between 2000 and 2002. In two cases multiple myeloma was discovered after the diagnosis of HIV infection. In the other case, the diagnosis of HIV infection was posterior to the occurrence of multiple myeloma. HIV infection was symptomatic in two cases who received consequently antiviral treatment. Multiple myeloma was diagnosed at an advanced stage in the three cases. The paraprotein was an IgG in two cases and an IgA in the other one. The CD4 counts before treatment were around 200/mm3 in two cases and within normal limits in the third case. Viral load was not measured. VMCP and VAMCP regimens were administered without major complications and under anti-infectious prophylaxis. The follow-up is still insufficient to assess the medium-term evolution and to determine the prognosis of multiple myeloma. The description of these three cases confirms the involvement of HIV in B cell lymphoma genesis. PMID- 15112899 TI - [Cannabis and schizophrenia. From euphoria to psychosis]. AB - Cannabis is the most abused illicit drug in the world. Prevalence of this substance abuse is more frequent in psychiatric populations, particularly in persons with schizophrenia. This observation leads one to question a relationship of cause and effect between cannabis and schizophrenia and of impact of cannabis on the course of this disease. Several studies have been realised to give a response to this question. PMID- 15112900 TI - [Peritoneal closure in abdominopelvic surgery: useful or not?]. AB - Peritoneal closure is a classical technique in abdominopelvic surgery. However, numerous studies demonstrated that omission of this surgical procedure did not lead to increased rate of short-term postoperative complications, and shortened the operative times. As a consequence, numerous surgeons deliberately choosed not to perform peritoneal closure. This manuscript updates about this topic. PMID- 15112901 TI - [How I explore ... diaper dermatitis]. AB - Diaper dermatitis is a frequent condition. Several clinical types are distinguished. The most frequent type results from increased fragility of the newborn buttock skin when covered by diapers. According to the mechanisms involved and the severity of the dermatitis, one can distinguish the intertrigo of the chubby baby, and the so-called "W", "Y" and "red panties" types of diaper dermatitis. When the effects of occlusion are not controlled by adequate absorption by the diapers, hyperhydration of the stratum corneum occurs and progresses to maceration. As a result, the value of the coefficient of friction of the skin increases with epidermal weakening to rubbing. In addition, fecal enzymes (urease, proteases and lipases) alter urines and skin. Judicious hygiene measures and the correct choice of diapers are mandatory. Cutaneous colonization by microorganisms, the most important of which being Candida albicans, is the main complication. Adequate preventive and curative measures can combat diaper dermatitis with confidence. A miconazole paste allows to improve the tribological properties of the interface between diapers and the skin. It also abates the impact of Candida albicans in the pathogenesis of the skin disorder. PMID- 15112902 TI - [Medication of the month. Insulin glargine (Lantus)]. AB - Insulin glargine (Lantus) is a human insulin analogue produced by recombinant DNA technology and recently launched by Aventis. Modification of the human insulin molecule at position A21 and at the C-terminus of the B-chain results in the formation of a stable compound that is soluble at pH 4.0, but forms amorphous microprecipitates in subcutaneous tissue (pH > 7,4) from which small amounts of insulin glargine are gradually released. The plasma concentration versus time profile of insulin glargine is therefore relatively constant over 24 hours as compared to conventional human insulins, especially NPH. This allows once-daily injection as basal insulin therapy, at any moment of the clock time (but if possible at the same time from day to day). Reproducibility of plasma insulin levels is also improved with insulin glargine as compared to human NPH insulin. Insulin glargine administration should be combined to rapid insulin injections, before each meal in order to control postprandial hyperglycaemia, or with oral antidiabetic agents in type 2 diabetes. The pharmacokinetic properties of insulin glargine allow an easier titration of basal insulin dose, which should facilitate adequate blood glucose control while decreasing the risk of hypoglycaemia, especially during night time. Insulin glargine use is safe with no increased antigenicity, immunogenicity or mitogenicity reactions as compared to human insulin. Optimal use of this new insulin analogue should be integrated in a global management of the diabetic patient as well as in a new culture of insulin therapy. PMID- 15112903 TI - Communicating with parents with full disclosure: a case of cloacal extrophy with genital ambiguity. AB - Full disclosure and complete involvement of parents in decisions concerning assignment of sex and genital surgery must be part of medical care for children presenting with findings consistent with disorders of intersex. Intersex most commonly involves disorders of steroidogenesis or gonadal function, but may include multiple cloacal anomalies, such as presented here. OBJECTIVE: To describe full disclosure of medical findings, by a multi-disciplinary medical team, as they became available over a period of weeks, in an infant originally assigned male, but eventually assigned female. PATIENT: An infant born at 24 weeks of gestation, after prenatal ultrasound showing distended bladder, ascites, and bilateral hydroureters, found to have an imperforate anus and a tubular structure appearing as a thin penis, without palpable corpora. DISCUSSION: Events concerning this case are discussed in relation to full disclosure of medical information to parents, guidelines for management of intersex, and the diagnosis (cloacal anomaly, cloacal extrophy, ano-rectal anomalies or uro-rectal septum malformation sequence). CONCLUSIONS: Full disclosure with involvement of parents with medical decisions is not only mandated currently, but also can be an effective approach in intersex care. PMID- 15112904 TI - The use of a continuous glucose monitoring system in hypoglycemic disorders. AB - OBJECTIVE: To evaluate the use of a continuous glucose monitoring system (CGMS) in the evaluation and treatment of infants and children with hypoglycemic disorders. METHODS: Patients with hypoglycemic disorders wore the CGMS device in the Pediatric Clinic Research Center during their evaluation and treatment. Capillary blood glucose (CBG) values were obtained at least 3 times each day and entered into the device for calibration purposes. We evaluated the number of hypoglycemic episodes below 3.3 mmol/l (60 mg/dl) detected by CGMS compared to CBG values and characterized episodes by their duration and intensity. RESULTS: Five patients with hypoglycemic disorders were included in the study. There were a total of 13,369 sensor points, 343 paired sensor and CBG data points, and 57 days included. A total of 180 episodes of hypoglycemia occurred in these five patients, with an average duration of 55 +/- 13 minutes. Using a cut-off of 3.3 mmol/l (60 mg/dl) for hypoglycemia, the sensor had a sensitivity of 65.4%, specificity of 90.6%, and false positive rate of 42.9%. The positive and negative predictive values were 57.1% and 93.2%, respectively. CONCLUSION: CGMS is a useful adjunct in the diagnosis and evaluation of hypoglycemia, and for documentation of euglycemia in these patients following therapy. PMID- 15112905 TI - Seasonality of month of birth among African American children with diabetes mellitus in the city of Chicago. AB - AIM: To study the seasonality of month of birth among African American children with insulin-treated diabetes mellitus (DM) in the city of Chicago, in order to determine whether perinatal exposures play a significant role in diabetes risk among children of non-European origin. METHOD: The Chicago Childhood Diabetes Registry ascertains new cases of insulin-treated DM among minority children < 18 years of age; these cases were compared with birth certificate data for the general African American population in Chicago. The chi2 test and Poisson regression were used to compare the pattern of month of birth of children with DM (n = 604) to that of the general population (n = 758,658) over the same period of years (1968-1995). RESULTS: In a month-by-month comparison, there were significantly fewer children who later developed DM born during October (chi2 = 6.74, df = 1). This seasonal pattern was stronger among males (n = 284) than females (n = 320), and among those who apparently developed type 2 DM (n = 155) compared to those who developed type 1 DM (n = 449). Children who were diagnosed between 15 and 17 years of age (n = 131) demonstrated significant seasonality (chi2 = 27.6, df = 11) compared to the general population. CONCLUSIONS: The apparent protective effect of October birth, and the significant overall seasonality among those diagnosed at ages 15-17 years, suggest the possibility that seasonal environmental factors at conception, during pregnancy or in the neonatal period may affect DM risk in adolescence. The greater impact of month of birth in adolescent type 2 DM patients is surprising and seems to indicate a role for mechanisms other than the immunological ones previously suggested. PMID- 15112906 TI - HLA-DQA1 and -DQB1 alleles in Latino and African American children with diabetes mellitus. AB - Few studies have described the genetics of childhood diabetes mellitus (DM) in US minorities. High-risk DQA1 and DQB1 alleles (DQA1*0301, DQB1*0201, and DQB1*0302 in African Americans and Latinos, and DQA1 *0501 in African Americans) were identified from previous studies and tested in 45 African American and 26 Latino patients from the population-based Chicago Childhood Diabetes Registry, and in 50 healthy race-matched controls. Sixteen of the African American patients and three Latinos had youth-onset type 2 DM and were analyzed separately. In African Americans with type 1 DM, both DQA1*0102 and DQB1*0602 were protective (p < 0.0001), and the susceptibility alleles DQA1*0301 and DQB1*0201 were more frequent than in controls (p < 0.01). In Latinos, DQA1*0301 and DQB1*0302 were marginally increased in patients with DM1 compared to controls; no individual DQA1 or DQB1 allele was protective. Patients with DM1 were significantly more likely to carry one or two high-risk DQA1 alleles in both populations; they were also more likely than controls to carry at least one high-risk DQB1 allele. The odds ratio for the ability to form at least two high-risk DQA1-DQB1 heterodimers (cis and/or trans) was 7.9 (95% CI: 1.7-40.0) for African Americans and 5.7 (1.3 25.6) for Latinos with DM1. African American patients with DM2 were not statistically different from controls, and were less likely to carry four high risk susceptibility alleles than patients with DM1 (p = 0.002). Many of the HLA DQ associations previously documented in non-Hispanic White populations also are found in African Americans and Latinos with DM1, although some differences exist. PMID- 15112907 TI - Efficacy of orlistat as an adjunct to behavioral treatment in overweight African American and Caucasian adolescents with obesity-related co-morbid conditions. AB - This pilot study compared the efficacy of orlistat as an adjunctive treatment for obesity between African American and Caucasian adolescents. Twenty obese adolescents with obesity-related co-morbid conditions underwent measurements of body composition, glucose homeostasis by frequently sampled intravenous glucose tolerance test (FSIGT), and fasting lipids before and after 6 months treatment with orlistat 120 mg tid in conjunction with a comprehensive behavioral program. Weight (p < 0.05), BMI (p < 0.001), total cholesterol (p < 0.001), LDL cholesterol (p < 0.001), fasting insulin (p < 0.02) and fasting glucose (p < 0.003) were lower after treatment. Insulin sensitivity, measured during the FSIGT, improved significantly (p < 0.02), as did fasting indices such as the homeostasis model assessment for insulin resistance (p < 0.01). African American subjects exhibited significantly less improvement in weight (p < 0.05), BMI (p < 0.01), waist circumference (p = 0.03), and insulin sensitivity (p = 0.05). Improvements in cholesterol were not significantly different between African Americans and Caucasians. We conclude that Caucasians lost more weight and had greater improvements in insulin sensitivity than African Americans, but both exhibited improvements in plasma lipids. The true benefit of orlistat treatment over a comprehensive behavioral program remains to be determined in placebo controlled trials. PMID- 15112908 TI - Protease inhibitor therapy improves protein catabolism in prepubertal children with HIV infection. AB - Past studies in adults have reported that loss of lean tissue mass (LTM) is associated with accelerated rates of protein catabolism. To date, studies of protein kinetics from pediatric patients infected by HIV have not been published; however, poor linear growth and weight loss are well-documented. The first aim of this study was to test the hypothesis that protein catabolism is high in pediatric patients with HIV. Protease inhibitors (PI) have proven to be effective therapy for pediatric HIV patients. One action of these drugs is that of lowering the viral burden, and several studies suggest that these drugs result in increased growth and weight velocity. Our second aim was to determine whether PI therapy improves protein catabolism. METHODS: We studied eight children infected with HIV (ages 2.9-6.2 years, Tanner stage I, CD4 counts 100,000-300,000, 5 F/3 M) and eight healthy age- and gender-matched controls. Measures of protein turnover were conducted using the stable isotope [1-(13C)]leucine. Body composition was measured by dual X-ray absorptiometry (DXA) scan for determination of LTM, and indirect calorimetry for measurement of resting energy expenditure. Children with HIV infection were studied at baseline and after 6 weeks of PI therapy; control children were studied only once. RESULTS: Protein catabolism, represented as leucine rate of appearance (Ra) in the fasted state, was higher in the HIV-infected children at baseline compared to control children. After 6 weeks of PI therapy, leucine Ra decreased, but not to the range found in control children. Leucine Ra correlated with viral burden. LTM significantly improved in all patients. CONCLUSION: These results suggest that similar to HIV infected adults, HIV-infected children have higher than normal protein catabolism. Furthermore, our measures suggest that short-term PI therapy results in improved protein catabolism and LTM. PMID- 15112909 TI - Homocysteine in a multi-ethnic sample of school-age children. AB - Cardiovascular disease is a major cause of morbidity and mortality in the United States and other developed countries; arterial lesions that are precursors of disease begin during childhood. Homocysteine levels have been associated with cardiovascular disease rates in adults, but information about levels in and impact on children is limited, particularly among various ethnic groups. This study examined the cardiovascular risk factors of a multi-ethnic sample of 100 9 15 year-old Native American, Hispanic, White, and mixed race children in rural central Washington. The mean fasting homocysteine level was 5.82 micromol/l (+/- 1.47), with no significant differences noted among ethnic groups. Mean dietary intake of folate, vitamin B6, and vitamin B12 exceeded current Recommended Dietary Allowances. Homocysteine levels did not show statistically significant correlations with cardiovascular risk factors. Homocysteine levels were not found to be a cardiovascular risk factor of importance, nor were significant ethnic differences found, in Native American, Hispanic and White children consuming adequate diets. PMID- 15112910 TI - The MACIS score predicts the clinical course of papillary thyroid carcinoma in children and adolescents. AB - MACIS (distant Metastasis, patient Age, Completeness of resection, local Invasion, and tumor Size) scores are employed to predict mortality for papillary thyroid carcinoma (PTC) in adults. However, this system has not been validated in children and adolescents. We hypothesized that MACIS scores would correlate with recurrent and persistent disease in children. Patients with PTC (n = 48) were divided into those with aggressive (invasive, metastasic, recurrent or persistent disease) or indolent (lacking these features) disease. Those with aggressive PTC (n = 11) had average MACIS score = 5.2 +/- 1.3, compared to 3.7 +/- 0.4 in patients with indolent disease (n = 37, p < 0.0005). A cutoff score of 4.0 provides a PPV of 29% and NPV of 94% in predicting recurrence, and a PPV of 43% and NPV of 91% in predicting persistent disease. MACIS scores may be useful in predicting outcome in the pediatric population. PMID- 15112911 TI - Catch-up growth in severe juvenile hypothyroidism: treatment with a GnRH analog. AB - Anecdotal reports suggest that the addition of a gonadotropin releasing hormone (GnRH) analog (GnRHa) in addition to L-thyroxine (LT4) replacement may increase adult stature in children with severe longstanding hypothyroidism by prolonging the pubertal growth period. This retrospective chart review compares the height outcome and body mass index in 33 children (21 treated with LT4 alone and 12 treated with LT4 + GnRHa) with severe longstanding hypothyroidism and bone age delay. Seventeen controls and six GnRHa-treated patients were followed to adult height (BA >14 yr [F]/16 yr [M] and/or growth velocity < 2 cm/yr). At diagnosis, GnRHa-treated patients were 1) older and shorter for chronological age, and 2) more advanced in puberty and bone age. Despite these differences, at adult height, both groups had similar improvements in height Z scores, similar height deficits, and comparable adult heights. Changes in BMI Z score were similar for both groups. Our study suggests that the addition of GnRHa to LT4 may improve interval growth without imposing a risk of obesity in children with longstanding severe hypothyroidism. PMID- 15112912 TI - Novel TSHbeta subunit gene mutation causing congenital central hypothyroidism in a newborn male. AB - Newborn screening programs that use only high TSH levels as a marker for hypothyroidism may overlook neonates with congenital hypothyroidism (CH) due to TSH deficiency. We sought the cause of TSH deficiency in a neonate with low levels of thyroxine and TSH. The coding region of the TSHbeta gene was amplified and its sequence examined for mutations. Two mutations in exon 3 were identified: 1) a nucleotide deletion of T410 in codon 105 resulting in a frameshift in one allele, and 2) a previously unreported nucleotide deletion of T266 in codon 57, causing a frameshift and a premature stop at codon 62 in the other allele. We describe a compound heterozygous patient with TSHbeta mutations at codons 57 and 105 that interfered with a critical disulfide bond in the TSH molecule and caused CH. State screening programs that measure both T4 and TSH levels have the potential to detect newborns with congenital central hypothyroidism. PMID- 15112913 TI - Diabetic ketoacidosis secondary to growth hormone treatment in a boy with Prader Willi syndrome and steatohepatitis. AB - A 13 year-old boy with Prader-Willi syndrome and steatohepatitis presented with diabetic ketoacidosis 4 weeks after the initiation of growth hormone (GH) treatment. He did not have signs or symptoms of type 2 diabetes mellitus (DM2) before the initiation of GH treatment. Hyperglycemia resolved 2 months after discontinuation of GH. He redeveloped DM2 6 months later associated with excessive weight gain. Diabetic ketoacidosis as a rare complication of GH therapy emphasizes the importance of screening for carbohydrate intolerance before and during GH treatment in patients with Prader-Willi syndrome. Steatohepatitis may be the only manifestation of insulin resistance and warrants further evaluation. PMID- 15112914 TI - McCune-Albright syndrome associated with pituitary microadenoma: patient report. AB - McCune-Albright syndrome (MAS) is a rare disorder characterized by the classic triad of precocious puberty, polyostotic fibrous dysplasia and cafe-au-lait spots. Additional endocrine abnormalities may also be present, including hyperthyroidism, growth hormone excess and hyperprolactinemia. The most commonly encountered endocrine dysfunction is gonadal hyperfunction. Gonadotropin independent precocious puberty is typically the initial manifestation of MAS in girls. Ovarian cysts may be detected on pelvic ultrasound. Our patient was also found to have pituitary microadenoma, evidenced by dynamic magnetic resonance imaging. PMID- 15112915 TI - Classical phenotype of Laron syndrome in a girl with a heterozygous mutation and heterozygous polymorphism of the growth hormone receptor gene. AB - We describe here a 19 month-old girl with classical Laron syndrome (LS). Molecular analysis of the GH receptor gene in the patient and her parents was performed. The patient was found to be heterozygous for a mutation in exon 4 (R43X) and heterozygous for a polymorphism in exon 6 (Gly168Gly). Her mother was also heterozygous for R43X but homozygous for the polymorphism. In the father, a heterozygous polymorphism was found. Contrary to previous assumptions that only homozygous patients express the typical phenotype, this patient shows all the classical features of LS, despite being a heterozygote for a pathological defect. PMID- 15112916 TI - The first meeting of the Loop Club. PMID- 15112917 TI - Dental hygienists and dental therapists. PMID- 15112918 TI - Why our ethics curricula don't work. AB - The impact our ethics curricula have on students seems marginal at best. Students take the ethics courses we offer and pass the tests we give, but no one's behavior changes as a result. We fundamentally see ourselves teaching about ethics, which is slightly different than teaching ethics--and expecting behavior to change as a result of what is taught. The premise of this article is that our ethics courses are inadequate in content and form to the extent that they do not cultivate an introspective orientation to professional life. In some cases they amount to little more than a study of various state dental practice acts or the Code of Ethics of the American Dental Association. Three specific weaknesses are identified in a typical ethics curriculum: 1) failure to recognize that more education is not the answer to everything; 2) ethics is boring; and 3) course content is qualitatively inadequate because it does not foster an introspective basis for true behavioral change. A fourth element, an innovation, is directed to this third weakness and entails implementing a precurriculum very early in the dental educational experience to address the disconnect between knowledge and action. PMID- 15112919 TI - Does performance on school-administered mock boards predict performance on a dental licensure exam? AB - Many dental schools consider the successful completion of a state or regional dental licensure examination as one of the significant benchmarks for assessing effectiveness of the curriculum. At the University of Florida College of Dentistry (UFCD), performance on the state dental licensure examination is monitored and compared with senior year mock board performance and clinical productivity to identify factors that may contribute to state board "pass" rates. A retrospective analysis was conducted of "first-time" performance on the Florida Dental Licensure Exam for graduates from classes 1996 to 2003. Using ANOVA, licensure exam performance data was analyzed and compared with performance on the senior mock board exam and clinical productivity, determined by numbers of procedures completed in each discipline. Significant relationships were noted between four of thirteen aspects of mock board performance and clinical productivity data and performance on the Florida Dental Licensure Exam. First, a significant relationship (p<0.05) was found between passing the senior mock board fixed prosthodontic preparation and successful completion of that procedure on the state licensure exam. Second, a significant relationship (p<0.05) was noted between the clinical (patient-based) Class II amalgam on the senior mock board and passing that procedure on the state licensure exam. Third, a significant relationship was noted (p<0.05) between the number of Class IV clinical composite procedures completed during dental school and passing the licensure exam Class IV manikin composite procedure. Fourth, there was a significant relationship (p<0.01) between the number of clinical Class II amalgam procedures completed during the junior and senior years and passing the state licensure exam clinical amalgam procedure. No significance was found between the remaining five mock board procedures (Class II composites, Class IV composites, pin amalgams, endodontic, and periodontal scaling/root planing) and performance on the like procedures on the licensure exam. Likewise, no significance was found between the remaining four productivity measures (numbers of Class II composites, endodontic teeth treated, crowns and abutments completed, and quadrants of periodontal scaling/root planing) and performance of these procedures on the state licensure exam. PMID- 15112920 TI - Financial management and dental school strength, Part I: Strategy. AB - The ultimate goal of financial management in a dental school is to accumulate assets that are available for strategic growth, which is a parallel objective to the profit motive in business. Budget development is often grounded in an income statement framework where the goal is to match revenues and expenses. Only when a balance sheet perspective (assets = liabilities + equity) is adopted can strategic growth be fully addressed. Four views of budgeting are presented in this article: 1) covering expenses, 2) shopping, 3) strategic support, and 4) budgeting as strategy. These perceptions of the budgeting process form a continuum, moving from a weak strategic position (covering expenses) to a strong one (budgeting as strategy) that encourages the accumulation of assets that build equity in the organization. PMID- 15112921 TI - Financial management and dental school equity, Part II: Tactics. AB - Financial management includes all processes that build organizations' equity through accumulating assets in strategically important areas. The tactical aspects of financial management are budget deployment and monitoring. Budget deployment is the process of making sure that costs are fairly allocated. Budget monitoring addresses issues of effective uses and outcomes of resources. This article describes contemporary deployment and monitoring mechanisms, including revenue positive and marginal analysis, present value, program phases, options logic, activity-based costing, economic value added, cost of quality, variance reconciliation, and balanced scorecards. The way financial decisions are framed affects comparative decision-making and even influences the arithmetic of accounting. Familiarity with these concepts should make it possible for dental educators to more fully participate in discussions about the relationships between budgeting and program strategy. PMID- 15112922 TI - Using significant event analysis in dental and medical education. AB - As part of a new professional development teaching strand at the University of Otago's School of Dentistry, final-year dental students were each asked to write a "thought-provoking episode report" (TPER). These TPERs formed the basis for group discussion in a professional development course. This article outlines the main content themes of the reports, comparing them with similar reports written by medical students from the same university. While both sets of reports demonstrate students' commitment to high standards of care, there were significant differences in the overall themes. Of the fifty-one dental TPERs, the main themes were "difficult" patients, receiving conflicting advice from different clinical tutors, friends as patients, belittlement, and maintenance of professional standards. Key themes from medical students' TPERs included responding to patient suffering, observing or experiencing belittlement, uncertainty, error and complaints, the role of the undergraduate student, treating family and friends, causing distress in order to learn, and issues with mentoring. Possible interpretations of the similarities and differences in this data include confirmation of the value of personal experience as a basis for student discussion, the necessity for formal programs in professional development, and the presence of a hidden curriculum in dental and medical education. PMID- 15112923 TI - A predoctoral clinical geriatric dentistry rotation at the University of the Pacific School of Dentistry. AB - Predoctoral dental students from University of the Pacific School of Dentistry provide initial and yearly dental evaluations for participants of On Lok Senior Health Services on site at On Lok centers. Student dentists also complete some dental procedures including denture fabrication, adjustments and repairs, hard and soft relines, scaling/root planing, polishing, and limited restorative treatments. A wide range of age-prevalent oral conditions such as candidiasis and xerostomia are identified and treated or managed. Students may also be called upon to present patient needs weekly to a member of the interdisciplinary team for discussion. Students periodically review instructions and devices for oral health care with the On Lok staff. The program is intended to be mutually beneficial to the participants of On Lok and Pacific student dentists. While the majority of comprehensive and emergency services are provided by On Lok staff dentists and contract specialists, the student dentist program has broadened the scope of the oral health program at On Lok and has been well integrated with the other day services. Meanwhile, Pacific students gain experience identifying and managing the complex social, economic, and health needs of frail elders in San Francisco. PMID- 15112924 TI - Developing cultural competence and social responsibility in preclinical dental students. AB - Dental student development of cultural competence and social responsibility is recognized by educators as an important element in the overall shaping of minds and attitudes of modem dental practitioners. Yet training modalities to achieve these competencies are not clearly defined, and outcome measurements are elusive. This article shows an effective method to meet these desired outcomes. Sixty-one freshmen (class of 2005) participated in forty hours of nondental community service, and reflective journals were completed by the end of second year. Competency outcomes were measured by selecting key words and phrases found in the individual journals. Key phrases were related to compassion, righteousness, propriety, and wisdom. Also, phrases had to be accompanied by written indications of direct program causation. The combination of active-learning (based upon service learning models) in public health settings outside of the dental realm, accompanied by reflective journaling, enhanced cultural understanding and community spirit in the majority of students. PMID- 15112926 TI - A profile of dental school deans, 2002. AB - Dental school deans were surveyed to update and expand the profile of a dean developed in a 1999 survey and to assess whether the profile has changed since the initial survey. The demographic characteristics and qualifications of dental school deans were similar in 1999 and 2002. The dean turnover rate and average length of deanship have not significantly changed since 1990. There was a shift in the current and emerging issues described by deans in the 1999 and 2002 surveys. In 2002, infrastructure and resource management, future workforce needs, and access to care were much more likely to be mentioned. Nearly all deans have a dental degree and additional advanced degrees or certificates, and they are likely to be specialists. Dental school deans are almost always full professors with tenure and have had significant administrative experience as department chairs and assistant/associate deans and have served on boards outside of their institution. Deans typically are scholars with multiple published journal articles, textbooks, or chapters and have been the principal investigator for grants and contracts. This information provides insight into the background necessary to become a dental school dean and the qualities sought by dental schools. PMID- 15112925 TI - Clinical simulation and foundation skills: an integrated multidisciplinary approach to teaching. AB - Faced with the challenge of restructuring a preclinical curriculum to provide a stronger background in general dentistry, a clinical simulation program that emphasizes critical thinking in clinical decision-making was developed and implemented at New York University College of Dentistry. The program offers an integrated program in clinical sciences focused on the faculty-defined outcomes for a general dentist. The curriculum was developed using outcomes that must be met before full patient care privileges are extended. The curriculum is centered around a series of patients with differing profiles of risk, disease, and treatment needs. Students are required to think globally, collect data that leads to an accurate assessment of the patient's risk, plan prevention and health promotion, and define a treatment plan. The student then demonstrates proficiency in executing treatment, evaluates the results, and speculates about the long-term impact of the treatment provided. Student and faculty evaluations are discussed as well as the strengths and shortcomings of the curriculum. PMID- 15112927 TI - High-throughput proteomics for alcohol research. AB - This report summarizes the proceedings of a satellite symposium of the 2003 Research Society on Alcoholism meeting held on June 21, 2003, in Fort Lauderdale, FL. The goal of this symposium, sponsored by the NIAAA, was to identify new proteomic directions in alcohol research that will (1) enable studies that focus on characterizing protein function, biochemical pathways, and networks to understand alcohol-related illnesses; (2) identify protein-protein interactions, posttranslational modifications, and subcellular localizations; (3) identify molecular targets for medication development; (4) develop biomarkers for susceptibility, dependence, consumption, and relapse, as well as alcohol-induced pathologies; and (5) develop high-throughput drug screens to test the efficacy of therapeutics that control alcohol-induced diseases. The purpose of the symposium was also to promote the application of high-throughput proteomic approaches, including isolation of membrane-bound proteins, in situ proteomics, large-scale two-dimensional separations, protein microarray platforms, mass spectrometry, matrix-assisted laser desorption/ionization, matrix-assisted laser desorption/ionization time-of-flight, liquid chromatography-tandem mass spectrometry, and isotope-coded affinity tags. In addition, the development of protein network maps by using new bioinformatics approaches for database mining was also discussed. PMID- 15112928 TI - Serotonin 5-HT2 receptors and alcohol: reward, withdrawal and discrimination. AB - This article represents the proceedings of a symposium at the 2003 Research Society on Alcoholism meeting in Fort Lauderdale, Florida. The organizer was Karl J. Buck, and the chairperson was Mark S. Brodie. The presentations were (1) The Multiple PDZ Domain Protein May Mediate Genetic Differences in Ethanol Withdrawal Severity Via Interaction With 5-HT2 Receptors, by Matthew T. Reilly and Kari J. Buck; (2) The Ionic Mechanism of Serotonin Potentiation of Ethanol Excitation of Ventral Tegmental Area Neurons, by Mark S. Brodie; and (3) 5-HT(2C) Receptor Agonists in the Discriminative Stimulus Effects of Ethanol, by Laura M. Rogers, Ken Szeliga, and Kathleen Grant. PMID- 15112929 TI - Ethanol effects on cell signaling mechanisms. PMID- 15112930 TI - The future of proteomics in the study of alcoholism. AB - This article represents the proceedings of a workshop at the 2003 annual meeting of the Research Society on Alcoholism in Fort Lauderdale, FL. The workshop organizers/chairpersons were Chinnaswamy Kasinathan and Paul Manowitz. The presentations were (1) Introduction to the field of proteomics, by Kent Vrana; (2) Use of proteomics in the identification of urinary biomarkers for alcohol intake, by Chinnaswamy Kasinathan, Paul Thomas, and Paul Manowitz; (3) Proteomics screening illuminates ethanol-mediated induction of HDL proteins in macaques, by Kent Vrana, Randy Gooch, Travis Worst, Stephen Walker, Aaron Xu, Peter Pierre, Heather Green, and Kathleen Grant; and (4) Proteomics applied to the study of the liver, by Laura Beretta. PMID- 15112931 TI - Consequences of multiple withdrawals from alcohol. AB - This article represents the proceedings of a symposium at the 2003 annual meeting of the Research Society on Alcoholism in Fort Lauderdale, FL, organized by Theodora Duka and chaired by Dai Stephens. The purpose of the symposium was to examine the effects of multiple experiences of withdrawal from alcohol in animals made dependent on alcohol and in humans who are alcohol dependent. Parallels were drawn to the effects of repeated short-lived high-content alcohol exposures in animals and in humans who are social drinkers but indulge in binge drinking. The presentations were (1) Multiple detoxifications and risk of relapse in abstinent alcoholics, by John Gentry and Robert Malcolm; (2) Emotional and cognitive impairments after long-term use of alcohol: relationship to multiple detoxifications and binge drinking, by Theodora Duka; (3) The effect of repeated withdrawal from ethanol on conditioning to appetitive stimuli, by Tamzin Ripley, Gilyanna Borlikova, and Dai Stephens; (4) Alcohol withdrawal kindling: electrographic measures in a murine model of behavioral seizure sensitization, by Lynn Veatch and Howard Becker; and (5) Binge drinking induced changes in CNS, by Fulton Crews. PMID- 15112932 TI - Biological and behavioral markers of alcohol sensitivity. AB - This article summarizes a symposium that was organized by Dr. Kim Fromme and presented at the 2003 annual meeting of the Research Society on Alcoholism in Ft. Lauderdale, Florida. The four presentations illustrate the emerging technologies and methods that are now being used to investigate the genetic basis of differential sensitivity to alcohol and their behavioral manifestations. Combining human genotyping with laboratory measures of behavior and subjective reports, these presentations represent state-of-the-art approaches to crossing the bridge from the Decade of the Brain to the Decade of Behavior. Dr. De Wit's paper describes her research on the neurobiological basis for individual differences in sensitivity to the stimulant and sedative effects of alcohol. Evidence suggests that activity of the dopaminergic and GABAergic neurotransmitters underlie these stimulant and sedative effects, respectively. Both Drs. Hutchison's and Corbin's papers describe their research on polymorphisms for the serotonin transporter (SLC6A4) as a determinant of the subjective effects of alcohol challenge. Dr. Hutchinson's and Ms. Ray's findings indicate that individuals with the short form of the SLC6A4 alleles (S) demonstrated a low level of response to alcohol, thus supporting previous research that the S allele may be associated with increased risk for alcohol dependence. In contrast, Dr. Corbin did not find a reliable association between the SLC6A4 genotype and subjective response to alcohol. Mr. Cook's and Dr. Wall's paper adds another dimension to this article by presenting research on both the aldehyde dehydrogenase (ALDH2) and alcohol dehydrogenase (ADH2) genetic variants and their association with the alcohol-related flushing response that is prevalent in Asian populations. Dr. David Goldman provides concluding remarks. PMID- 15112933 TI - Serotonin-3 receptors in the actions of alcohol, alcohol reinforcement, and alcoholism. AB - This article represents the proceedings of a symposium at the 2003 annual meeting of the Research Society on Alcoholism in Fort Lauderdale, FL. The organizers and chairs were William J. McBride and David M. Lovinger. The presentations were (1) Mechanisms of alcohol potentiation of 5-HT3 receptor function, by David M. Lovinger and Tina Machu; (2) Chronic alcohol drinking alters 5-HT3 receptors regulating the mesolimbic dopamine system, by Richard J. Thielen; (3) 5-HT3 receptors in the VTA regulate alcohol drinking and the reinforcing effects of alcohol, by Zachary A. Rodd and James M. Murphy; and (4) Ondansetron as a treatment for "biological" alcoholism, by John D. Roache and Bankole A. Johnson. PMID- 15112934 TI - Integrative neurobiology of the alcohol withdrawal syndrome--from anxiety to seizures. AB - This article represents the proceedings of a symposium presented at the 2003 Research Society on Alcoholism meeting in Ft. Lauderdale, Florida, organized and chaired by Carl L. Faingold. The presentations were (1) Overview, by Carl L. Faingold; (2) Stress, Multiple Alcohol Withdrawals, and Anxiety, by Darin Knapp; (3) Relationship Between Genetic Differences in Alcohol Drinking and Alcohol Withdrawal, by Julia Chester; (4) Neuronal Mechanisms in the Network for Alcohol Withdrawal Seizures: Modulation by Excitatory Amino Acid Receptors, by Carl L. Faingold; and (5) Treatment of Acute Alcohol Withdrawal and Long-Lasting Alterations in Hippocampal Neuronal Networks, by Larry P. Gonzalez. The presentations emphasized the importance of using intact behaving animals to advance the understanding of the human alcohol withdrawal syndrome. This involves applying and amplifying the neurophysiological and neurotransmitter findings observed in vitro to the network-based neurobiological mechanisms that are involved in several important aspects of the specific behaviors observed clinically. The symposium provided evidence that the organizational aspects of neuronal networks in the intact nervous system add another nexus for the action of alcohol and drugs to treat alcohol withdrawal that may not be readily studied in isolated neural elements used in in vitro approaches. PMID- 15112935 TI - Building bridges: the transdisciplinary study of craving from the animal laboratory to the lamppost. AB - This article represents the proceedings of a symposium at the 2003 Research Society on Alcoholism meeting in Ft. Lauderdale, Florida, organized and chaired by Peter M. Monti. The presentations and presenters were (1) Alcohol Seeking and Self-Administration in Rats: The Role of Serotonin Activity, by Cristine L. Czachowski; (2) Assessing Binge Drinking in Monkeys, by Kathleen A. Grant; (3) Craving and the Perception of Time, by Michael Sayette; (4) Ecological and Laboratory Assessment of Alcohol Urges and Drinking: Effects of Naltrexone, by Peter M. Monti; and (5) Discussion, by Damaris J. Rohsenow. PMID- 15112936 TI - Overlapping peptide control of alcohol self-administration and feeding. AB - This article represents the proceedings of a symposium at the 2003 annual meeting of the Research Society on Alcoholism in Fort Lauderdale, FL. The organizers and chairpersons were Mark Egli and Todd E. Thiele. The presentations were (1) Voluntary alcohol consumption is modulated by central melanocortin receptors, by Todd E. Thiele; (2) Central infusion of neuropeptide Y reduces alcohol drinking in alcohol-preferring P rats, by Robert B. Stewart and Nancy E. Badia-Elder; (3) The gut peptide cholecystokinin controls alcohol intake in Sardinian alcohol preferring rats, by Nori Geary and Maurizio Massi; and (4) Hypothalamic galanin: a possible role in excess alcohol drinking, by Sarah F. Leibowitz and Bartley G. Hoebel. PMID- 15112937 TI - Development of novel pharmacotherapies for the treatment of alcohol dependence: focus on antiepileptics. AB - This article represents the proceedings of a symposium, "Development of Novel Pharmacotherapies for the Treatment of Alcohol Dependence: Focus on Antiepileptics," presented at the 2003 annual meeting of the Research Society on Alcoholism in Fort Lauderdale, FL. The organizers and cochairs were Bankole A. Johnson and Robert M. Swift. The presentations were (1) Development of topiramate in the treatment of alcoholism, by Bankole A. Johnson; (2) Craving as a predictor of treatment outcome in pharmacotherapy trials for the treatment of alcoholism, by Nassima Ait-Daoud; (3) Use of biomarkers as a predictor of treatment response in treating alcoholism, by Martin A. Javors; (4) Psychotherapy to enhance compliance with pharmacotherapy in treatment trials for alcohol dependence, by Carlo C. DiClemente; (5) Synopsis of promising medications to treat alcoholism, by Robert M. Swift; and (6) New knowledge on the development of antiepileptic medications for the treatment of alcoholism, by Robert J. Malcolm, Jr. PMID- 15112938 TI - Pharmacological treatment of alcohol abuse/dependence with psychiatric comorbidity. AB - This article represents the proceedings of a symposium at the 2003 annual meeting RSA in Fort Lauderdale, FL. It was organized and cochaired by Charlene E. Le Fauve and Carrie L. Randall. The presentations were (1) Introduction, by Charlene E. Le Fauve and Raye Z. Litten; (2) Treatment of co-occurring alcohol use and anxiety disorders, by Carrie L. Randall and Sarah W. Book; (3) Pharmacological treatment of alcohol dependent patients with comorbid depression, by Darlene H. Moak; (4) Efficacy of valproate in bipolar alcoholics: a double blind, placebo controlled study, by Ihsan M. Salloum, Jack R. Cornelius, Dennis C. Daley, Levent Kirisci, Johnathan Himmelhoch, and Michael E. Thase; (5) Alcoholism and schizophrenia: effects of antipsychotics, by Alan I. Green, Robert E. Drake, Suzannah V. Zimmet, Rael D. Strous, Melinda Salomon, and Mark Brenner; and (6) Conclusions, by Charlene E. Le Fauve; discussant, Raye Z. Litten. Alcohol dependent individuals have exceptionally high rates of co-occurring psychiatric disorders. Although this population is more likely to seek alcoholism treatment than noncomorbid alcoholics, the prognosis for treatment is often poor, particularly among patients with more severe psychiatric illnesses. Development of effective interventions to treat this population is in the early stages of research. Although the interaction between the psychiatric condition and alcoholism is complex, progress has been made. The NIAAA has supported a number of state-of-the-art pharmacological and behavioral trials in a variety of comorbid psychiatric disorders. Some of these trials have been completed and are presented here. The symposium presented some new research findings from clinical studies with the aim of facilitating the development of treatments that improve alcohol and psychiatric outcomes among individuals with alcohol-use disorders and co-occurring psychiatric disorders. The panel focused on social anxiety disorder, depression, bipolar disorder, and schizophrenia. PMID- 15112939 TI - The Veterans Aging Cohort Study: observational studies of alcohol use, abuse, and outcomes among human immunodeficiency virus-infected veterans. AB - This article represents the proceedings of a symposium at the 2003 annual meeting of the Research Society on Alcoholism in Fort Lauderdale, FL. The organizers/chairs were Joseph Conigliaro and Amy Justice. The presentations were (1) Introduction, by Joseph Conigliaro and Tamra Madenwald; (2) Alcohol and HIV/AIDS: the importance of integrative and translational research, by Kendall Bryant; (3) Alcohol use and abuse among patients with HIV infection, by Joseph Conigliaro and Stephan Maisto; (4) Severity of comorbid alcohol use/abuse in HIV infection, by Amy Justice and Jeffrey Samet; (5) Estimating the impact of alcohol use on long-term HIV outcomes, by Scott Braithwaite and Amy Justice; (6) Homelessness, drug & alcohol use among HIV+ veterans, by Adam Gordon and Robert Cook; and (7) Hepatitis C & alcohol in the VACS 3 study, by Shawn Fultz and Kevin Kraemer. The symposium concluded with a discussion led and facilitated by Diedra Roach. PMID- 15112940 TI - Approaches to brief intervention for hazardous drinking in young people. AB - This article represents the proceedings of a symposium at the 2002 joint conference of the Research Society on Alcoholism and the International Society for Biomedical Research on Alcoholism in San Francisco. The chair was John B. Saunders. The focus of the symposium was on brief intervention approaches for hazardous drinking among young people. The presentations were (1) Evidence for the effectiveness of brief intervention as an approach to reducing hazardous alcohol use, by John B. Saunders; (2) College student hazardous drinking in New Zealand, the USA, UK, and Australia: implications for research, policy, and intervention, by Kypros Kypri; (3) Applications of motivational feedback on the college campus, by Scott T. Walters; (4) A population based individualized alcohol harm reduction feedback intervention: preliminary results from the college-based alcohol risk reduction (CBARR) trial, by Robert G. Laforge; and (5) Brief interventions: conclusions and future directions, by Mary E. Larimer. PMID- 15112941 TI - Collecting longitudinal data through childhood, adolescence, and young adulthood: methodological challenges. AB - This article presents the proceedings of a workshop at the 2003 Research Society on Alcoholism meeting in Fort Lauderdale, Florida. The organizers and chairs were Vivian Faden and Nancy Day. The presentations were (1) Lessons Learned From the Lives Across Time Longitudinal Study, by Michael Windle and Rebecca Windle; (2) Methodological Issues in Longitudinal Surveys With Children and Adolescents, by Joel Grube; (3) The Pittsburgh ADHD Longitudinal Study: Methodological and Conceptual Challenges, by Brooke Molina, William Pelham, Elizabeth Gnagy, and Tracey Wilson; and (4) Lessons learned in Conducting Longitudinal Research on Alcohol Involvement: If Only I Had Known Before Hand! by Kristina Jackson and Kenneth Sher. PMID- 15112942 TI - Really underage drinkers: alcohol use among elementary students. AB - Despite the current societal concern with underage drinking, little attention has been paid to alcohol use within the preadolescent population. This article presents the proceedings of a symposium held at the 2003 Research Society on Alcoholism meeting in Fort Lauderdale, Florida, that was organized and chaired by John E. Donovan. The intent of the symposium was to kick start research on alcohol use among elementary school children by reviewing what is known regarding drinking in childhood. Presentations included (1) The Epidemiology of Children's Alcohol Use, by John E. Donovan; (2) The Validity of Children's Self-Reports of Alcohol Use, by Sharon L. Leech; (3) Predicting Onset of Drinking From Behavior at Three Years of Age: Influence of Early Child Expectancies and Parental Alcohol Involvement Upon Early First Use, by Robert A. Zucker; and (4) Parent, Peer, and Child Risk Factors for Alcohol Use in Two Cohorts of Elementary School Children, by Carol J. Loveland-Cherry. Presentations indicated the need for better nationwide surveillance of children's experience with alcohol; suggested that children's reports of their use of alcohol tend to be reliable and valid; supported children's alcohol use schemas and parental drinking and alcoholism at child age three as independent predictors of early onset drinking; and showed that onset of drinking before fourth or fifth grade, peer pressure, and parental norms and monitoring predict elementary student alcohol use and misuse. PMID- 15112943 TI - Alcohol-induced neurodegeneration: when, where and why? AB - This manuscript reviews the proceedings of a symposium organized by Drs. Antonio Noronha and Fulton Crews presented at the 2003 Research Society on Alcoholism meeting. The purpose of the symposium was to examine recent findings on when alcohol induced brain damage occurs, e.g., during intoxication and/or during alcohol withdrawal. Further studies investigate specific brain regions (where) and the mechanisms (why) of alcoholic neurodegeneration. The presentations were (1) Characterization of Synaptic Loss in Cerebella of Mature and Senescent Rats after Lengthy Chronic Ethanol Consumption, (2) Ethanol Withdrawal Both Causes Neurotoxicity and Inhibits Neuronal Recovery Processes in Rat Organotypic Hippocampal Cultures, (3) Binge Drinking-Induced Brain Damage: Genetic and Age Related Effects, (4) Binge Ethanol-Induced Brain Damage: Involvement of Edema, Arachidonic Acid and Tissue Necrosis Factor alpha (TNFalpha), and (5) Cyclic AMP Cascade, Stem Cells and Ethanol. Taken together these studies suggest that alcoholic neurodegeneration occurs through multiple mechanisms and in multiple brain regions both during intoxication and withdrawal. PMID- 15112944 TI - [Sheltering: whose responsibility is it? An experience report]. AB - This paper is about the sheltering of users in a basic health unit in Porto Alegre/RS according to the experience during the Integrated Residency Program in Collective Health at the Public Health School and reflections about specific bibliography. Sheltering is the manner in which the health service is organized to attend to its users and to answer their demands, relying on health education processes aiming to build a different relation than that established in "emergency rooms". It is believed that the nurse is the professional indicated for this task because of its basic education in collectivity and integralized human care. PMID- 15112945 TI - [Ethnography and ethnonursing: research methods in nursing]. AB - This paper aims to review the literature about the qualitative ethnographic research method, and its application in nursing, describing one of the authors experience while writing a master's dissertation. Both methods, ethnography and ethnonursing, are presented with their specific proposals and phases. Leininger's ideas are enhanced since it was this author who revealed to nursing the importance to comprehend culture, allowing to give care which is coherent to each ones beliefs and values. The study allowed to better know the research methods and it is wished that it contributes to deepen the knowledge about them, stimulating their use. PMID- 15112946 TI - [The child with a cancer diagnosis: revisiting the way of assistance policies]. AB - This paper considers about the onco-hematologic assistance policies and their implications in caring for a child with cancer diagnosis and his/her family. It emphasizes that the existential changes of the child-family unit may be minimized by the policies that assure access to resources. The study shows how Non governmental Organizations (NGOs) are inserted in the context. The conclusion is that the engagement of families as well of health professionals and institutions, both governmental and non-governmental, may contribute to the implementation of policies that enlarge the network of assistance to children who suffer from an onco-hematologic disease. PMID- 15112947 TI - [Patterns of knowledge in nursing and their implications in teaching]. AB - The paper presents nursing as a discipline that is still setting its boundaries and that because of this has the need to define its own form of knowledge and how to produce and validate it. With this in mind, a brief literature review on knowledge patterns which are specific to nursing, as well as empirical, ethical, esthetical and personal patterns are presented. From the characterization of each one of these patterns, its implication in teaching is discussed, highlighting the need to privilege those forms of knowledge during the process of formal teaching learning. PMID- 15112948 TI - [Health and work: a correlation of concepts in the perspectives of a rural population and of Christophe Dejours]. AB - This study deepens the discussion about the concept of health constructed by Christophe Dejours and the one perceived by families who are descended from Italian immigrants who are agricultural workers at the Fourth Colony of Italian Immigration, Rio Grande do Sul, Brazil. In this paper, the concept of health is discussed viewing the academic and popular knowledge. As the study develops, it is possible to perceive the occurrence of an approximation of the author's thoughts with the families descending from Italian immigrants regarding health and work. Among the elements that confirm this closeness is the fact that only the human being himself is able to establish the limits between normal and pathological. PMID- 15112949 TI - [The software QSR Nvivo 2.0 in qualitative data analysis: a tool for health and human sciences researches]. AB - The QSR Nvivo 2.0 is one of the latest versions of qualitative data analysis software package. Taking on board our experience as QSR Nvivo 2.0 users we describe here the software's most important tools. The QSR Nvivo 2.0 was used to facilitate the qualitative analysis of data gathered in a Health and Education research. Considering the shortage of published materials about the issue, which suggests a lack of knowledge about the program in our context, our aim is to show how the QSR Nvivo 2.0 can assist qualitative data analysis. PMID- 15112950 TI - [Hospital escort program for the geriatric patient]. AB - It is presented a report of nine years of experience with the Hospital Escort Program for geriatric patients, carried out by nurses of a University Hospital (UFSC), at the Medical Unit II, since 1993. The goal is to integrate the family that cares for the elderly patient, bringing them to the hospital as a Nursing partner or client. The program also seeks to give psychosocial support to the patients; to enhance the escort's skills to care for the elderly on their daily activities and also on specific health tasks after hospital discharge; to develop healthy life practices within the family, which had shown appreciation and recognition to the program. PMID- 15112951 TI - [How do asthmatic women experience the illness?]. AB - This study aimed to identify and understand the essence of the anguish of five asthmatic women, patients of the Pneumology Service of a hospital in Porto Alegre, and how they interact with the health professionals. It uses the qualitative methodology, with semi-structured interviews. The content analysis identified: a dependent relationship and an incidence of separation anguish demonstrated by the way the women deal with medication and the caregivers; some of the women's impediments; difficulties to describe the illness and asthma crisis. The separation anxiety and the dependent behavior of the patient must be understood by the caregiver. PMID- 15112952 TI - [Sexually active women's beliefs in choosing or rejecting contraceptive methods]. AB - We have interviewed 23 women from December 2000 to April 2001 at a Public Health Unit on a Family Health Program, aiming to analyze their beliefs regarding the use or not of contraceptive methods, and to identify their central or peripheral axis. 123 beliefs were emitted and categorized in five types, each one of them classified in 11 structural units, representing rationales for supporting the beliefs. There was a tendency to centralize the beliefs, with a worrisome 47.15% of the women pertaining to type B: unwilling to accept changes. There is a need to discuss beliefs and partners as part of educational programs. PMID- 15112953 TI - [The history of psychiatric nursing in Rio Grande do Sul: Part I]. AB - The objective of this article is to contribute towards the history and historiography of Psychiatric Nursing in Rio Grande do Sul by reconstituting the Professional School of Nursing that functioned from 1939 to 1952 at the Sao Pedro Hospital. Since this is recent history, documentary research and oral history were used as primary sources of information. It was possible to characterize the context that favored its organization, the main characteristics of the process used to train nurses in the field of psychiatry and its effects in attaining the discipline of these professionals in relation to the psychiatric project, which at that time was hegemonic. PMID- 15112954 TI - [Nursing procedure: a dimension of the communication with the newborn]. AB - This article approaches the perceptions of the nursing staff about the procedure's role in communicating with the newborn. Such theme was identified in a qualitative, exploratory/descriptive study developed with the nursing staff of the neonatal unit of an University Hospital in Porto Alegre/RS. The article explores the procedure's implications to the caregiver and methodological aspects of the study. It approaches the caregiver's perceptions about the communication that is due to skin stimuli; the feelings involved in performing the procedure; the relation between the inflicted pain and communication; as well as the opportunity to teach and learn through the procedure. PMID- 15112955 TI - SF2809 compounds, novel chymase inhibitors from Dactylosporangium sp. 1. Taxonomy, fermentation, isolation and biological properties. AB - Six novel chymase inhibitors, SF2809-I, SF2809-II, SF2809-III, SF2809-IV, SF2809 V and SF2809-VI, were isolated from the fermentation broth of an actinomycete strain SF2809. The strain was identified as Dactylosporangium sp. by morphological, chemotaxonomical and phylogenetic studies. These six novel compounds inhibited recombinant human chymase in the range between IC50 of 0.014 and 7.3 microM. However, they showed little or no inhibitory activity against chymotrypsin or cathepsin G, even though these two and chymase belong to the chymotryptic serine protease family. This result indicates that these compounds work as specific chymase inhibitors. PMID- 15112956 TI - SF2809 compounds, novel chymase inhibitors from Dactylosporangium sp. 2. Structural elucidation. AB - Novel chymase inhibitors, SF2809-I, II, III, IV, V and VI, were isolated from the fermentation broth of Dactylosporangium sp. SF2809, and their structures were determined by spectroscopic analyses. SF2809 compounds commonly contain a substituted indole moiety and a quinolinone moiety. The two moieties are connected to a methylene carbon in SF2809-I and III. The other compounds, SF2809 II, IV, V and VI, have an additional moiety, a p-hydroxyphenyl group or a phenyl group. In these compounds, all of three moieties are connected to a methine carbon. Furthermore, studies concerning the stereochemistry of SF2809-V revealed that the isolated compound was racemic, and the isomer possessing (R) configuration was about thirty times more potent than another isomer. PMID- 15112957 TI - ICM0301s, new angiogenesis inhibitors from Aspergillus sp. F-1491. I. Taxonomy, fermentation, isolation and biological activities. AB - In the course of screening program for inhibitors of angiogenesis, novel substances designated as ICM0301A approximately H (1 approximately 8) were isolated from the culture broth of Aspergillus sp. F-1491. ICM0301s inhibited the growth of human umbilical vein endothelial cells (HUVECs) induced by basic fibroblast growth factor (bFGF) with IC50 values of 2.2 approximately 9.3 microg/ml. ICM0301A (1) showed significant anti-angiogenic activity at lower than 10 microg/ml in the angiogenesis model using rat aorta cultured in fibrin gel. ICM0301s showed very low cytotoxicity against various tumor cells. Furthermore, 1CM0301A did not show any toxic symptom in mice by intraperitoneal injection at 100 mg/kg. PMID- 15112958 TI - ICM0301s, new angiogenesis inhibitors from Aspergillus sp. F-1491. II. Physico chemical properties and structure elucidation. AB - ICM0301A (1), B (2) and their congeners (3 approximately 8) were isolated from a culture broth of Aspergillus sp. F-1491 as new angiogenesis inhibitors. Their structures were elucidated by spectroscopic analyses. ICM0301A and B have a substituted decalin skeleton containing two oxirane rings. PMID- 15112959 TI - New beauvericins, potentiators of antifungal miconazole activity, Produced by Beauveria sp. FKI-1366. I. Taxonomy, fermentation, isolation and biological properties. AB - Three new beauvericins, designated beauvericins D, E and F, were isolated along with known beauvericin and beauvericin A, from the culture of Beauveria sp. FKI 1366 by solvent extraction, ODS column chromatography and HPLC. These compounds potentiate miconazole activity against not only wild Candida albicans but also fluconazole resistant C. albicans. Beauvericins D and E decreased the IC50 value of miconazole against fluconazole resistant C. albicans from 1.3 microM to 0.25 and 0.31 microM, respectively. PMID- 15112960 TI - New beauvericins, potentiators of antifungal miconazole activity, Produced by Beauveria sp. FKI-1366. II. Structure elucidation. AB - The structures of beauvericins D, E and F, novel potentiators of miconazole activity against Candida albicans produced by Beauveria sp. FKI 1366, were elucidated by various spectroscopic analyses including UV, NMR, and MS and degradation experiments. They have the common skeleton of the 18-membered cyclodepsipeptides. PMID- 15112961 TI - Isolation and structural characterization of siderophores, madurastatins, produced by a pathogenic Actinomadura madurae. AB - Madurastatins Al (1), A2 (2) and A3 (3), novel pentapeptides that were acylated with salicylic acid at the N-terminus, were isolated from the culture broth of a pathogenic Actinomadura madurae IFM 0745 strain. These structures were mainly determined by 2D NMR and MS/MS spectral techniques. The strain produced simultaneously madurastatins B1 (4) and B2 (5) consisting of Ser and salicylic acid moieties. Compounds 1 and 4 had an antibacterial activity against Micrococcus luteus, indicating that the presence of the aziridine ring is essential for such activity. Because 1 has a strong affinity with ferric ion due to the presence of two hydroxamic acids and a salicylic acid, it is considered to be a siderophore that is a low molecular weight iron chelater. The production of siderophores may be one of the characteristics of pathogenic microorganisms. PMID- 15112962 TI - Exploitation of heparanase inhibitors from microbial metabolites using an efficient visual screening system. AB - In this paper we describe the establishment of an efficient visual method for screening heparanase inhibitors, and we present the results of screening 10,000 microbial culture broths. Heparanase-overexpressing stable clones of the human hepatocellular carcinoma HepG2 cells were established and used as an enzyme source. Digestion of heparan sulfate (HS) was detected using novel HS-containing tablets or SDS-polyacrylamide gel electrophoresis. This method was able to find suramin, a known heparanase inhibitor, from a library of typical enzyme inhibitors. By screening 10,000 culture broths of microorganisms (actinomycetes, fungi, and bacteria) an actinomycete strain, RK99-A234, was found to have heparanase inhibitory activity. RK-682 was identified in the fermentation broth as a heparanase inhibitor, IC50 = 17 microM. PMID- 15112963 TI - Carminomycin, 14-hydroxycarminomycin and its novel carbohydrate derivatives potently kill human tumor cells and their multidrug resistant variants. AB - The new hydrophilic derivatives of 14-hydroxycarminomycin were obtained using 13 dimethyl ketal of 14-bromocarminomycin (6) as the starting compound. The reductive alkylation of 6 with melibiose or D-galactose followed by hydrolysis of the corresponding intermediate bromoketals 9 and 11 produced 3'-N-[-alpha-D (galactopyranosyl-(1 --> 6)-O-D-1-desoxyglucit-1-yl]-14-hydroxycarminomycin (10) and 3'-N-(1-desoxy-D-galactit-1-yl)-14-hydroxycarminomycin (12), respectively. These novel derivatives 10 and 12 were less toxic than carminomycin or 14 hydroxycarminomycin for leukemia (K562) and breast carcinoma (MCF-7) cells. Importantly, carminomycin, 14-hydroxycarminomycin and compounds 10 and 12 were similarly active for wild type cells and their multidrug resistant (MDR) sublines, K562i/S9 and MCF-7Dox. PMID- 15112964 TI - Ajudazols, new inhibitors of the mitochondrial electron transport from Chondromyces crocatus. Production, antimicrobial activity and mechanism of action. PMID- 15112965 TI - Structure revision of FD-891, a 16-membered macrolide antibiotic. PMID- 15112966 TI - Laryngeal findings in advanced Parkinson's disease. AB - Parkinson's disease is a major source of neurologic morbidity. A majority of patients with Parkinson's disease complain of problems with voice, speech, and swallowing. Treatments for these problems center on the improvement of vocal fold adduction through either speech therapy or vocal fold augmentation. No prior study has looked at laryngeal improvement after neurologic surgery, specifically deep brain stimulation, performed to treat Parkinson's disease. The goal of this study was to establish a baseline of laryngeal findings in patients who are considering deep brain stimulation. Fifteen patients underwent physical examination with videostroboscopy and fiberoptic endoscopic evaluation of swallowing before deep brain stimulation. In addition, they were asked to self report voice handicap. Eighty-seven percent of patients demonstrated significant vocal fold bowing. All patients had some degree of pharyngeal residue of solids noted on evaluation of swallowing. All but one patient had a significant self reported voice handicap. These findings are reviewed and established as a baseline for further study. PMID- 15112967 TI - Injection laryngoplasty with calcium hydroxylapatite gel implant in an in vivo canine model. AB - The ideal injectable agent for vocal fold medialization is biocompatible, durable, sized to prevent phagocytosis and migration, and formulated for easy injection and does not adversely affect the viscoelastic properties of the vocal fold. We tested a cohesive implant of calcium hydroxylapatite (CaHA) particles in a gel carrier in an in vivo canine model of phonation. Six dogs underwent unilateral recurrent laryngeal nerve section and injection laryngoplasty of the paralyzed vocal fold with a CaHA implant. The six follow-up examinations were performed at 1, 2, 3, 6, 9, and 12 months, and the larynx and bilateral neck lymphatic system were harvested for histologic analysis. The CaHA implant adequately medialized the vocal fold to regain glottal closure. The mucosal waves remained unaltered from baseline. The implant remained soft in the larynx and did not migrate to the neck lymphatic system. A localized foreign body giant cell reaction was present on histologic evaluation, but not acute or other chronic inflammation. A size analysis revealed no resorption of the CaHA particles. A decrease in medialization was noted at all follow-up intervals related to resorption of the aqueous-based gel carrier. The CaHA implant appears to be relatively safe and suitable for injection laryngoplasty. PMID- 15112968 TI - Office-based treatment of glottal dysplasia and papillomatosis with the 585-nm pulsed dye laser and local anesthesia. AB - Treatment of glottal papillomatosis and dysplasia was mirror-guided and performed in surgeons' offices in the 19th century. It migrated to the operating room in the 20th century to accommodate direct laryngoscopic surgery, which required assistants to administer anesthesia and procedural support. Presently, the primary treatment goals, which are disease regression and voice restoration or maintenance, are tempered by the morbidity of general anesthesia and potential treatment-induced vocal deterioration. In fact, general anesthesia has been appropriately considered to be an acceptable source of morbidity for the promise of a precise procedure, which usually ensures airway safety and an optimal vocal outcome. However, patients with recurrent glottal papillomatosis and keratosis with dysplasia are typically monitored with various degrees of watchful waiting until there is a subjective judgment (on the part of the patient and surgeon) that the disease is more of a liability than is the procedure to treat it. Innovations in the 585-nm pulsed dye laser delivery system have allowed for its use in the clinic with local anesthesia through the working channel of a flexible fiberoptic laryngoscope. A prospective assessment was done on 51 patients in 82 cases of recurrent glottal papillomatosis (30) and dysplasia (52). All individuals had previously undergone microlaryngoscopic management with histopathologic evaluation. Five procedures could not be completed because of impaired exposure (2) or discomfort (3). Of those patients who could be treated, there was at least a 50% disease involution in 68 of 77 cases (88%) and 25% to 50% disease regression in the remaining 9 (12%). Patient self-assessment of the voice revealed that 34 of 77 were improved, 39 were unchanged, 4 were slightly worse, and none were substantially worse. These data confirm that diseased mucosa can be normalized without resection or substantial loss of vocal function. The putative mechanisms, which vary according to the fluence (energy) delivered by the laser, are photoangiolysis of sublesional microcirculation. denaturing of epithelial basement membrane linking proteins, and cellular destruction. Furthermore, this relatively safe, effective technique allowed for treatment of many patients (in a clinic setting) in whom classic surgery-related morbidity would have often delayed intervention. PMID- 15112969 TI - Vocal efficiency measurements in subjects with vocal polyps and nodules: a preliminary report. AB - Vocal efficiency is a quantitative measure of the ability of the larynx to convert subglottal power to acoustic power. On the basis of the scant previous literature and clinical intuition, we tested the hypothesis that vocal efficiency, as an indicator of the functional status of the larynx, is abnormally reduced in persons with vocal nodules and polyps. Because the most difficult aspect of obtaining measures of vocal efficiency has been the determination of subglottal pressure, we applied a noninvasive airflow interruption technique for this purpose. Subjects with normal voices (n = 22), vocal polyps (n = 14), and vocal nodules (n = 16) phonated at different intensities into a mask connected by way of piping to a flow meter, a pressure transducer, and an acoustic microphone. Inflation of a balloon-type valve located within the piping provided interruption of phonation. The intraoral pressure plateau occurring during flow interruption was used to estimate subglottal pressure. Subglottal power and acoustic power were determined, and their quotient provided a measure of vocal efficiency. The vocal efficiency in the normal subjects averaged 1.15 x 10(-5) at 70 dB, 3.17 x 10(-5) at 75 dB, 7.52 x 10(-5) at 80 dB, and 1.41 x 10(-4) at 85 dB. The vocal efficiency in the patients with vocal polyps averaged 3.62 x 10(-6) at 70 dB, 8.34 x 10(-6) at 75 dB, 2.10 x 10(-5) at 80 dB, and 4.26 x 10(-5) at 85 dB. The vocal efficiency in the patients with vocal nodules averaged 4.32 x 10(-6) at 70 dB, 1.57 x 10(-5) at 75 dB, 4.26 x 10(-5) at 80 dB, and 8.34 x 10(-5) at 85 dB. As compared to the normal subjects, the patients with laryngeal polyps or vocal nodules had significantly reduced vocal efficiency. These results provide quantitative verification of the clinical impression of inefficient phonation in patients with mass lesions of the vocal folds. PMID- 15112970 TI - Roof of the parapharyngeal space: defining its boundaries and clinical implications. AB - The roof of the parapharyngeal space (PPS) is poorly defined. Although it is generally described as having prestyloid and poststyloid compartments, we believe that these terms are imprecise. Therefore, we define its boundaries, partition, and compartments. We completed macroanatomical and microanatomical dissections in 10 specimens from 5 human cadaver heads; bone measurements in 50 dry skulls; and axial and coronal cross-sectional studies in 2 cadaveric specimens. The PPS roof is bordered laterally by the medial pterygoid fascia and medially by the pharyngobasilar fascia. The tensor veli palatini fascia (TVPF) partitions this roof into an anterolateral compartment containing fat and part of the deep lobe of the parotid gland, and a posteromedial compartment containing the cartilaginous part of the eustachian tube, internal carotid artery, internal jugular vein, and cranial nerves IX through XII. The anteroposterior length measures 32 mm (range, 26.1 to 36.9 mm), and the mediolateral width measures 16.3 mm (range, 12.1 to 21.3 mm). The PPS roof has 3 important bony landmarks (ie, scaphoid fossa, styloid process, sphenoid spine); 3 important fasciae (ie, medial pterygoid fascia, TVPF, pharyngobasilar fascia); and 2 compartments, which are anterolateral and posteromedial to the TVPF. We believe that this is the first report to specifically focus on the roof of the PPS. PMID- 15112971 TI - Pediatric airway reconstruction: principles, decision-making, and outcomes at the University of Iowa hospitals and clinics. AB - Pediatric airway stenosis challenges the treating surgeon, from the initial evaluation and decision-making process to the final postoperative care and follow up setting. As our approach to these cases evolves, we must critically evaluate our outcomes in reference to our stated goals. We describe our process in treating this population and critically examine our outcome data from 1990 to the present. Emphasis is placed on selection of procedure and postoperative management. PMID- 15112972 TI - Sphenoid sinus mucocele with unilateral blindness. AB - Mucoceles of the sphenoid sinus are rare and may frequently remain undiagnosed until symptoms due to compression of surrounding structures set in. In such cases, the optic nerve, because of its close proximity to the sphenoid sinus, is at risk, and pressure exerted by the mucocele may result in visual impairment. A case of a sphenoid sinus mucocele with sudden-onset unilateral blindness as the only presenting symptom has been described. Early diagnosis and prompt surgical intervention enabled the vision to be restored completely. PMID- 15112973 TI - Association between polymorphisms of the angiotensin-converting enzyme and angiotensinogen genes and allergic rhinitis in a Korean population. AB - Angiotensin-converting enzyme (ACE) inactivates bradykinin, substance P, and neurokinin A, which are thought to play important roles in the pathogenesis of inflammatory diseases. Expression of angiotensinogen, a precursor of angiotensin, is enhanced by augmented secretion of proinflammatory cytokines (eg, interleukin 1) in the site of inflammation. Insertion or deletion (I/D) ACE and M235T angiotensinogen gene polymorphisms were reported to be associated with atopy in a Czech population. Using polymerase chain reaction restriction fragment length polymorphism and SNaPshot typing analysis, we investigated the frequencies of the genotypes and alleles of the ACE gene in 137 patients with allergic rhinitis, of the M235T angiotensinogen gene in 186 patients with allergic rhinitis, and of both in 219 healthy control subjects. There was no difference in the frequency of the DD genotype of the ACE gene in the controls and patients (odds ratio, 1.32 [0.66-2.60]; p > .05). The D allele was more frequent in patients, but the difference was not statistically significant (odds ratio, 1.21 [0.89-1.64]; p > .05). There was no difference in the frequency of the TT genotype of the angiotensinogen gene in the controls and patients (odds ratio, 1.01 [0.38-2.69]; p > .05). The T allele was more frequent in patients, but the difference was not statistically significant (odds ratio, 1.10 [0.78-1.55]; p > .05). Our results indicate that polymorphisms in the genes for ACE and angiotensinogen may not be related to the development of allergic rhinitis in the Korean population. PMID- 15112974 TI - Surgical application of a new robotic system for paranasal sinus surgery. AB - The applicability of a robotic system for fully automated surgical procedures approaching the sphenoid sinus is evaluated. An integrated robotic system, A73, for computer navigation-guided, fully automated, and telemanipulation robotic performance is described. Details of the system comprising newly designed surgical instruments for robotic operations and preoperative planning protocols are provided. Experiments with an operational accuracy of less than 1 mm were followed by surgical tests, in which the results of fully automated and telemanipulation performances on 5 cadaveric heads are seen. The A73 system has been successfully used for a reproducible and accurate resection of the anterior wall of the sphenoid sinus. Therefore, we conclude that this system is suited for further testing toward approaching fully automated and more complex procedures of paranasal surgery. PMID- 15112975 TI - Clinical ototoxicity of teicoplanin. AB - No strong evidence of ototoxicity of teicoplanin can be found in the literature, possibly because of conservative definitions of ototoxicity. We performed audiometry over time to compare the ototoxicity of teicoplanin with that of cloxacillin as a non-ototoxic standard. The data were analyzed with a linear mixed-effects model. The hearing thresholds of 12 patients who were treated with teicoplanin for severe staphylococcal infections showed a slight but significant increase over time, whereas the thresholds of 5 patients treated with cloxacillin decreased significantly during treatment. This improvement in hearing with cloxacillin may be attributed to improvement of the clinical condition. This outcome implies that previous reports that suggest a lack of ototoxicity of teicoplanin potentially underestimate the risk and should be interpreted accordingly. PMID- 15112976 TI - Electronystagmographic findings in benign paroxysmal positional vertigo. AB - The aim of this study was to investigate the electronystagmographic findings in patients with benign paroxysmal positional vertigo. A retrospective review of the records of 168 patients with this disease during the past 3 years was performed. Epidemiological data and results from the audiological and neuro-otologic workup, including electronystagmography, were recorded. One hundred fifty-one patients had involvement of the posterior canal, 14 of the horizontal canal, and 3 of the anterior canal. Seventy-two patients (42.8%) had abnormal findings on the caloric tests. Thirty-seven of them (22%) had canal paresis and 23 (13.7%) had directional preponderance, whereas in 12 patients (7.1%) both unilateral weakness and directional preponderance were found. Finally, 21 patients (12.5%) had spontaneous nystagmus. It may be thus concluded that electronystagmographic abnormalities are quite common in patients with benign paroxysmal positional vertigo. Their presence may be explained according to several mechanisms, which are further discussed herein. PMID- 15112977 TI - Enlarged translabyrinthine approach for the management of large and giant acoustic neuromas: a report of 175 consecutive cases. AB - The translabyrinthine approach was once considered inadequate for the removal of acoustic neuromas (ANs), but that theory has few proponents today. Over the years, the translabyrinthine approach has been modified into the enlarged translabyrinthine approach, with experience and technical refinements leading to a wider access. Between April 1987 and December 2001, the Gruppo Otologico of Piacenza-Rome was able to remove 175 ANs 3 cm or larger in size from the cerebellopontine angle by adopting this modified surgical technique. These tumors represented 24.7% of all 707 ANs for which surgery was performed during the same period of time. Among the 175 cases, there was only 1 death. The incidence of complications was very low and was comparable to results previously published in the literature. Consequently, the hospital stay was short, with a mean of 7.3 days (5.1 days in the last 45 cases). The preoperative ipsilateral hearing was already compromised in 119 of the 175 cases (68%; class C/D according to the Committee on Hearing and Equilibrium of the American Academy of Otolaryngology Head and Neck Surgery, 1995). From our results, we can conclude that the use of the enlarged translabyrinthine approach in AN surgery is not dependent on tumor size. On the contrary, the advantages of a low rate of morbidity and a short hospital stay are ample proof that this is the best approach for the removal of large ANs. PMID- 15112978 TI - "Spontaneous" perilymph fistula: a case report. AB - A surgically documented case of barotrauma-induced perilymph fistula is presented in this case report. A brief review of the literature on this entity confirms the difficulty of making a definitive preoperative diagnosis in most instances. Clinical, audiometric, radiologic, and intraoperative findings are presented, and the classic presumed mechanisms for this uncommon cause of sudden sensorineural hearing loss are discussed. The presence of intact evoked otoacoustic emissions in an ear demonstrating a severe cochlear-type loss was considered helpful in narrowing the differential diagnosis in this case, and may suggest a productive avenue for future study. PMID- 15112979 TI - Database for the collection and analysis of clinical data and images of neoplasms of the sinonasal tract. AB - The Neoplasms of the Sinonasal Tract software package (NSNT v 1.0) implements a complete visual database for patients with sinonasal neoplasia, facilitating standardization of data and statistical analysis. The software, which is compatible with the Macintosh and Windows platforms, provides multiuser application with a dedicated server (on Windows NT or 2000 or Macintosh OS 9 or X and a network of clients) together with web access, if required. The system hardware consists of an Apple Power Macintosh G4500 MHz computer with PCI bus, 256 Mb of RAM plus 60 Gb hard disk, or any IBM-compatible computer with a Pentium 2 processor. Image acquisition may be performed with different frame-grabber cards for analog or digital video input of different standards (PAL, SECAM, or NTSC) and levels of quality (VHS, S-VHS, Betacam, Mini DV, DV). The visual database is based on 4th Dimension by 4D Inc, and video compression is made in real-time MPEG format. Six sections have been developed: demographics, symptoms, extent of disease, radiology, treatment, and follow-up. Acquisition of data includes computed tomography and magnetic resonance imaging, histology, and endoscopy images, allowing sequential comparison. Statistical analysis integral to the program provides Kaplan-Meier survival curves. The development of a dedicated, user-friendly database for sinonasal neoplasia facilitates a multicenter network and has obvious clinical and research benefits. PMID- 15112981 TI - Dr Arthur Proetz gave an address entitled "How to smother an idea". PMID- 15112980 TI - Hyoid bone fracture. PMID- 15112982 TI - Tools for Nanoscience at McGill University: sometimes smaller is better. PMID- 15112983 TI - Grafting miniature DNA binding proteins. AB - Miniature proteins serve as leads for biological and medicinal applications by positioning all amino acids necessary for biomolecular recognition on a compact protein structure. Protein grafting was recently used to create miniature helical proteins with high DNA binding affinity and specificity. PMID- 15112984 TI - Don't classify polyketide synthases. AB - Polyketide synthases are intensively studied as metabolite factories generating diverse biologically active natural products. Contrary to their current classification as different "types," there is now a growing body of evidence illustrating that nature realized limitless transitional stages during evolution. PMID- 15112985 TI - Breaking up is easy to do (if you're a DNA enzyme that cleaves RNA). AB - In this issue of Chemistry & Biology, Cruz et al. use in vitro selection to select deoxyribozymes that collectively cleave almost any RNA dinucleotide junction. More remarkable is the finding that the new enzymes are related to the 8-17 deoxyribozyme that cleaves AG dinucleotide junctions. PMID- 15112986 TI - Aureolic acids: similar antibiotics with different biosynthetic gene clusters. AB - In this issue of Chemistry & Biology, Mendez and colleagues describe the sequence and organization of the chromomycin gene cluster. Unexpectedly, the arrangement is starkly different from the mithramycin biosynthetic cluster, despite similarity in the individual genes and the near identical structures of the two antibiotic aureolic acids. PMID- 15112987 TI - Unraveling the pathway of lipoic acid biosynthesis. AB - Lipoic acid is almost universally required for aerobic metabolism. However, the mechanism for its synthesis and incorporation into proteins has remained elusive. A groundbreaking study published in the December issue of Chemistry & Biology uncovers critical features of the lipoic acid biosynthetic pathway. PMID- 15112988 TI - Profound insights into squalene cyclization. AB - In this issue of Chemistry & Biology, our understanding of the formation of pentacyclic hopene from the linear squalene is enhanced by an X-ray structure of a complex between squalene-hopene cyclase and the substrate analog 2-azasqualene. PMID- 15112989 TI - New probes for microtubule dynamics. AB - A phenotype-based screen identifies a purine analog, named diminutol, that perturbs the microtubule cytoskeleton in cells. An affinity-based approach identifies a protein target of this small molecule and leads to the characterization of a new pathway that may regulate cytoskeleton dynamics. PMID- 15112990 TI - Targeting FOXO kills two birds with one stone. AB - PTEN deficiency activates Akt signaling and results in a variety of human malignancies. Encouragingly, recent studies demonstrate that small molecules can regulate FOXO1a, an Akt target, to suppress tumor growth, and FOXO1a is therefore a promising anticancer drug target. PMID- 15112991 TI - Orthogonal base pairs continue to evolve. AB - Recent developments in the design and construction of unusual analogs of the natural nucleic acid bases have reached a milestone with the report (in this issue of Chemistry & Biology) of a new orthogonal base pair that allows site specific introduction of a photo-crosslinkable modified base into an RNA molecule by T7 RNA polymerase-mediated transcription of DNA containing the base-pairing partner. PMID- 15112992 TI - Biosynthesis of the antitumor chromomycin A3 in Streptomyces griseus: analysis of the gene cluster and rational design of novel chromomycin analogs. AB - The biosynthetic gene cluster of the aureolic acid type antitumor drug chromomycin A3 from S. griseus subsp. griseus has been identified and characterized. It spans 43 kb and contains 36 genes involved in polyketide biosynthesis and modification, deoxysugar biosynthesis and sugar transfer, pathway regulation and resistance. The organization of the cluster clearly differs from that of the closely related mithramycin. Involvement of the cluster in chromomycin A3 biosynthesis was demonstrated by disrupting the cmmWI gene encoding a polyketide reductase involved in side chain reduction. Three novel chromomycin derivatives were obtained, named chromomycin SK, chromomycin SA, and chromomycin SDK, which show antitumor activity and differ with respect to their 3 side chains. A pathway for the biosynthesis of chromomycin A3 and its deoxysugars is proposed. PMID- 15112993 TI - Leinamycin biosynthesis revealing unprecedented architectural complexity for a hybrid polyketide synthase and nonribosomal peptide synthetase. AB - A 135,638 bp DNA region that encompasses the leinamycin (LNM) biosynthetic gene cluster was sequenced from Streptomyces atroolivaceus S-140. The boundaries of the lnm cluster were defined by systematic inactivation of open reading frames within the sequenced region. The lnm cluster spans 61.3 kb of DNA and consists of 27 genes encoding nonribosomal peptide synthetase (NRPS), polyketide synthase (PKS), hybrid NRPS-PKS, resistance, regulatory, and tailoring enzymes, as well as proteins of unknown function. A model for LNM biosynthesis is proposed, central to which is the LNM hybrid NRPS-PKS megasynthetase consisting of discrete (LnmQ and LnmP) and modular (LnmI) NRPS, acyltransferase-less PKS (LnmG, LnmI, and LnmJ), and PKS modules with unusual domain organization. These studies unveil an unprecedented architectural complexity for the LNM hybrid NRPS-PKS megasynthetase and set the stage to investigate the molecular basis for LNM biosynthesis. PMID- 15112994 TI - Site-specific incorporation of a photo-crosslinking component into RNA by T7 transcription mediated by unnatural base pairs. AB - A photo-sensitive ribonucleotide of 5-iodo-2-oxo(1H) pyridine (Iy) capable of site-specific incorporation into transcripts was developed. The site-specific Iy incorporation into RNA was achieved by T7 transcription mediated by unnatural base pairing between Iy and its partner, 2-amino-6-(2-thienyl)purine (s). By this specific transcription, Iy was incorporated into an anti(Raf-1) RNA aptamer, which binds to human Raf-1 and inhibits the interaction between Raf-1 and Ras. Protein-dependent photo-dimerization of the aptamer was observed when Iy was located at specific positions in the aptamer, showing that the site-specific incorporation of the photo-sensitive component into RNA achieves highly specific crosslinking. This specific transcription mediated by the unnatural base pair would be a powerful tool for generating high-affinity RNA ligands and for analyzing RNA-RNA and RNA-protein interactions, as well as for constructing RNA based nanostructures. PMID- 15112995 TI - Dinucleotide junction cleavage versatility of 8-17 deoxyribozyme. AB - We conducted 16 parallel in vitro selection experiments to isolate catalytic DNAs from a common DNA library for the cleavage of all 16 possible dinucleotide junctions of RNA incorporated into a common DNA/RNA chimeric substrate sequence. We discovered hundreds of sequence variations of the 8-17 deoxyribozyme--an RNA cleaving catalytic DNA motif previously reported--from nearly all 16 final pools. Sequence analyses identified four absolutely conserved nucleotides in 8-17. Five representative 8-17 variants were tested for substrate cleavage in trans, and together they were able to cleave 14 dinucleotide junctions. New 8-17 variants required Mn2+ to support their broad dinucleotide cleavage capabilities. We hypothesize that 8-17 has a tertiary structure composed of an enzymatic core executing catalysis and a structural facilitator providing structural fine tuning when different dinucleotide junctions are given as cleavage sites. PMID- 15112996 TI - Controlling the DNA binding specificity of bHLH proteins through intramolecular interactions. AB - Reversible control of the conformation of proteins was employed to probe the relationship between flexibility and specificity of the basic helix-loop-helix protein MyoD. A fusion protein (apaMyoD) was designed where the basic DNA binding helix of MyoD was stablized by an amino-terminal extension with a sequence derived from the bee venom peptide apamin. The disulfide-stabilized helix from apamin served as a nucleus for a helix that extended for a further ten residues, thereby holding apaMyoD's DNA recognition helix in a predominantly alpha-helical conformation. The thermal stability of the DNA complexes of apaMyoD was increased by 13 degrees C relative to MyoD-bHLH. Measurements of the fluorescence anisotropy change on DNA binding indicated that apaMyoD bound to E-box-containing DNA sequences with enhanced affinity relative to MyoD-bHLH. Consequently, the DNA binding specificity of apaMyoD was increased 10-fold. PMID- 15112997 TI - Cloning, sequencing, and functional analysis of the biosynthetic gene cluster of macrolactam antibiotic vicenistatin in Streptomyces halstedii. AB - Vicenistatin, an antitumor antibiotic isolated from Streptomyces halstedii, is a unique 20-membered macrocyclic lactam with a novel aminosugar vicenisamine. The vicenistatin biosynthetic gene cluster (vin) spanning approximately 64 kbp was cloned and sequenced. The cluster contains putative genes for the aglycon biosynthesis including four modular polyketide synthases (PKSs), glutamate mutase, acyl CoA-ligase, and AMP-ligase. Also found in the cluster are genes of NDP-hexose 4,6-dehydratase and aminotransferase for vicenisamine biosynthesis. For the functional confirmation of the cluster, a putative glycosyltransferase gene product, VinC, was heterologously expressed, and the vicenisamine transfer reaction to the aglycon was chemically proved. A unique feature of the vicenistatin PKS is that the loading module contains only an acyl carrier protein domain, in contrast to other known PKS-loading modules containing certain activation domains. Activation of the starter acyl group by separate polypeptides is postulated as well. PMID- 15112998 TI - Biosynthesis of the angiogenesis inhibitor borrelidin by Streptomyces parvulus Tu4055: cluster analysis and assignment of functions. AB - The biosynthetic gene cluster for the angiogenesis inhibitor borrelidin has been cloned from Streptomyces parvulus Tu4055. Sequence analysis indicates that the macrolide ring of borrelidin is formed by a modular polyketide synthase (PKS) (borA1-A6), a result that was confirmed by disruption of borA3. The borrelidin PKS is striking because only seven rather than the nine modules expected for a nonaketide product are encoded by borA1-A6. The starter unit of the PKS has been verified as trans-cyclopentane-1,2-dicarboxylic acid (trans-1,2-CPDA), and the genes involved in its biosynthesis identified. Other genes responsible for biosynthesis of the nitrile moiety, regulation, and self-resistance were also identified. PMID- 15112999 TI - Design, synthesis, and characterization of urokinase plasminogen-activator sensitive near-infrared reporter. AB - The urokinase-type plasminogen activator (uPA) plays a critical role in malignancies, and its overexpression has been linked to poor clinical prognosis in breast cancer. The ability to noninvasively and serially map uPA expression as a biomarker would thus have significant potential in improving novel cancer therapies. Here, we describe the development of a selective uPA activatable near infrared (NIR) fluorescent imaging probe. The probe consists of multiple peptide motifs, GGSGRSANAKC-NH2, terminally capped with different NIR fluorochromes (Cy5.5 or Cy7) and a pegylated poly-L-lysine graft copolymer. Upon addition of recombinant human uPA to the probe, significant fluorescence amplification was observed, up to 680% with the optimized preparation. No activation with negative control compounds and uPA inhibitors could be measured. These data indicate that the optimized preparation should be useful for imaging uPA in cancer. PMID- 15113000 TI - Biosynthetic gene cluster of the glycopeptide antibiotic teicoplanin: characterization of two glycosyltransferases and the key acyltransferase. AB - The gene cluster encoding biosynthesis of the clinically important glycopeptide antibiotic teicoplanin has been cloned from Actinoplanes teichomyceticus. Forty nine putative open reading frames (ORFs) were identified within an 89 kbp genetic locus and assigned roles in teicoplanin biosynthesis, export, resistance, and regulation. Two ORFs, designated orfs 1 and 10*, showed significant homology to known glycosyltransferases. When heterologously expressed in Escherichia coli, these glycosyltransferases were shown to catalyze the transfer of UDP-(N-acetyl) glucosamine onto, respectively, 3-chloro-beta-hydroxytyrosine-6 (3-Cl-6betaHty) and 4-hydroxyphenylglycine-4 (4Hpg) of the teicoplanin heptapeptide aglycone. The product of another ORF, orf11*, was demonstrated in vitro to transfer n-acetyl-, n-butyryl-, and n-octanoyl-groups from acyl-CoA donors either to a free UDP aminosugar or to an aminosugar moiety in the teicoplanin pseudoaglycone, thus identifying Orf11* as the key acyltransferase in teicoplanin maturation. These findings should accelerate the combinatorial engineering of new and improved glycopeptide drugs. PMID- 15113001 TI - Conversion of squalene to the pentacarbocyclic hopene. AB - The membrane protein squalene-hopene cyclase was cocrystallized with 2 azasqualene and analyzed by X-ray diffraction to 2.13 A resolution. The conformation of this close analog was clearly established, and it agreed with the common textbook presentation. The bound squalene undergoes only small conformational changes during the formation of rings A through D, thus requiring no intermediate. However, ring E formation is hindered by an entropic barrier, which may explain its absence in the steroids. The structure analysis revealed a mobile region between the active center cavity and the membrane, which may melt, opening a passage for squalene and hopene. PMID- 15113002 TI - Binding of high-mannose-type oligosaccharides and synthetic oligomannose clusters to human antibody 2G12: implications for HIV-1 vaccine design. AB - Human antibody 2G12 broadly neutralizes human immunodeficiency virus type 1 (HIV 1) isolates and shows protective activity against viral challenge in animal models. Previous mutational analysis suggested that 2G12 recognized a novel cluster of high-mannose type oligosaccharides on HIV-1 gp120. To explore the carbohydrate antigen for HIV-1 vaccine design, we have studied the binding of 2G12 to an array of HIV-1 high-mannose type oligosaccharides by competitive ELISAs and found that Man9GlcNAc is 210- and 74-fold more effective than Man5GlcNAc and Man6GlcNAc in binding to 2G12. The results establish that the larger high-mannose oligosaccharide on HIV-1 is the favorable subunit for 2G12 recognition. To mimic the putative epitope of 2G12, we have created scaffold based multivalent Man9 clusters and found that the galactose-scaffolded bi-, tri , and tetra-valent Man9 clusters are 7-, 22-, and 73-fold more effective in binding to 2G12 than the monomeric Man9GlcNAc2Asn. The experimental data shed light on further structural optimization of epitope mimics for developing a carbohydrate-based HIV-1 vaccine. PMID- 15113004 TI - Myocardial contrast echocardiography: advantages of quantitative analysis. PMID- 15113003 TI - Identification of a novel protein regulating microtubule stability through a chemical approach. AB - To identify novel proteins regulating the microtubule cytoskeleton, we screened a library of purine derivatives using mitotic spindle assembly in Xenopus egg extracts as an assay. Out of a collection of 1561 compounds, we identified 15 that destabilized microtubules without targeting tubulin directly, resulting in small spindles. Affinity chromatography with one compound, named diminutol, revealed a potential target as NQO1, an NADP-dependent oxidoreductase. A role for NQO1 in influencing microtubule polymerization was confirmed through inhibition studies using known inhibitors and immunodepletion. Therefore, this chemical approach has identified a novel factor required for microtubule morphogenesis and cell division. PMID- 15113005 TI - Doppler echocardiographic assessment of coronary artery disease: a challenge becomes reality. PMID- 15113006 TI - Fantastic voyage through the cardiovascular system. PMID- 15113007 TI - Improved interpretation of dobutamine stress echocardiography following 4 months of systematic training in patients following acute myocardial infarction. AB - OBJECTIVES: Interpretation of stress echocardiography is subjective, and highly dependent on the experience of the interpreter. We sought to evaluate whether a cardiologist without any previous experience in stress echocardiography could adequately learn the skills of interpreting dobutamine stress echocardiograms (DSE) in post-infarct patients, after a period of systematic training. METHODS: A trainee in cardiology blindly reported 51 consecutive DSEs from a database of post-infarction studies, after 2 and 4 months of systematic training. We compared his interpretation with that of an expert. RESULTS: Agreement between the trainee and the expert improved significantly from 2 to 4 months of training in the left anterior descending artery territory for the overall scan interpretation (from kappa = 0.58 to kappa = 0.73; p = 0.03), wall thickening assessment in individual segments (from kappa = 0.40 to kappa = 0.55; p < 0.01) and the diagnosis of viable myocardium (from kappa = 0.11 to kappa = 0.43; p = 0.01). Similar improvement was observed in left circumflex, but not in the right coronary artery territory. Agreement in identifying inducible ischaemia also remained poor. CONCLUSION: This study suggests that systematic training can significantly reduce interobserver variability in a short time frame (4 months) and may improve the interpretation of DSE by a trainee. But improvements in image quality and use of predefined reading criteria are necessary to improve interobserver agreement further in myocardial regions where conformity in dobutamine stress echocardiographic interpretation is low. PMID- 15113008 TI - Comparison of left atrial size by freehand scanning three-dimensional echocardiography and two-dimensional echocardiography. AB - AIMS: As the left ventricular (LV) dimension is a poor indicator of LV volume, there are well-known limitations of left atrial (LA) antero-posterior dimensions as indicators of chamber size. LA volume has been shown to provide a more accurate assessment of LA size than LA dimension. To evaluate two-dimensional (2D)-derived LA volume in assessing LA size, we compared LA dimension and 2D LA volume with three-dimensional (3D)-derived LA volume. METHODS: We performed transthoracic freehand scanning 3D echocardiography (3D EchoTech, Germany) using magnetic fields and a harmonic imaging system in 32 patients. We collected a series of LA tomograms by slowly tilting the probe (fan-like scanning) in the parasternal position. The 3D LA volume was calculated by using the multiplanar Simpson's method. The 2D LA volume was measured by using the modified biplane Simpson's rule. RESULTS: LA antero-posterior dimensions and 2D volumes showed a significant positive correlation with 3D LA volumes. However, the correlation coefficient was significantly greater for the relationship between 2D LA volumes and 3D LA volumes than for that between LA dimensions and 3D LA volume. CONCLUSIONS: The 2D LA volumes provide a more accurate measure of the true size of the LA and are more sensitive to changes in LA size. PMID- 15113009 TI - Simultaneous transesophageal Doppler assessment of coronary flow reserve in the left anterior descending artery and coronary sinus allows differentiation between proximal and non-proximal left anterior descending artery stenoses. AB - AIM AND METHODS: The role of simultaneous transesophageal Doppler assessment of coronary flow reserve (CFR) in the left anterior descending artery (LAD) and coronary sinus (CS) in the diagnostics of hemodynamically significant LAD stenoses of various localization was studied in 16 CAD patients with angiographically proven >50% stenotic atherosclerosis of the LAD (nine--in the proximal third, seven--in the mid and/or distal third) and 23 healthy volunteers (all men). Dipyridamole was used as a stress agent. The diastolic phase of coronary flow in the LAD and the antegrade phase of coronary flow in the CS were analyzed. CFR in the LAD and CS was calculated in two ways: one--as ratio of peak hyperemic flow velocity to the peak baseline blood flow velocity (CFR by Vp); two -as ratio of volume hyperemic blood flow velocity to the volume baseline blood flow velocity (CFR by VBF). The level of the CFR <2 in both ways of calculation was diagnosed as reduced. RESULTS: It was found that in CAD patients with LAD proximal stenosis the values of CFR in the LAD were significantly lower than those in healthy individuals by both Vp (1.87 +/- 0.43 and 3.54 +/- 0.82; P<0.001) and VBF (1.79 +/- 0.77 and 3.85 +/- 1.25; P<0.01). In proximal stenosis CFR in the LAD by Vp was significantly lower than that in non-proximal stenosis (1.87 +/- 0.43 and 3.31 +/- 1.44; P<0.05). Sensitivity and specificity of CFR <2 in the LAD by Vp in the diagnostics of LAD proximal stenosis were 56% and 97%, respectively; and CFR <2 in the LAD by VBF--89% and 93%, respectively. In CAD patients with both proximal and non-proximal LAD stenoses CFR in the CS by Vp was significantly lower than that in healthy volunteers and was 1.74 +/- 0.53, 1.63 +/- 0.30 and 2.56 +/- 0.87; P<0.05, respectively. Sensitivity and specificity of CFR <2 in the CS by Vp in the diagnostics of hemodynamically significant LAD stenoses were 75% and 70%, respectively. The values of CFR in the CS by VBF in CAD patients and healthy volunteers did not differ significantly. CONCLUSIONS: Thus, simultaneous evaluation of CFR in the LAD and CS makes it possible to diagnose hemodynamically significant LAD stenoses and to differentiate between proximal and non-proximal impairments. PMID- 15113010 TI - Radiofrequency catheter ablation of ventricular tachycardia guided by intracardiac echocardiography. AB - AIMS: Ventricular tachycardia (VT) frequently has an anatomical substrate. Identification of areas prone to arrhythmogenicity facilitates radiofrequency catheter ablation (RFCA). Furthermore, direct monitoring of complications potentially increases safety of RFCA. The aim of this study was to evaluate the feasibility of guiding RFCA of VT with intracardiac echocardiography (ICE), in order to improve outcome and procedural safety. METHODS AND RESULTS: Eleven patients (age 59 +/- 15 years) with drug-refractory VT of various etiologies were studied. VT mapping and ablation were performed using standard techniques. ICE was performed with a multifrequency (5-10 MHz) phased-array transducer positioned in the right ventricle. Twenty different VTs were treated (CL 352 +/- 120 ms, 2.0 +/- 0.9 VT per patient). LV a- or dyskinesia was identified in all post-infarct patients. In patients with arrhythmogenic right ventricular dysplasia, right ventricular aneurysms and dyskinesia could be identified. In all patients catheter position and tip-tissue contact could easily be monitored with ICE. Procedural success (non-inducibility of hemodynamically stable VT) was achieved in all patients. Complications did not occur. CONCLUSION: ICE is feasible in guiding RFCA of VT of different etiologies. The use of ICE in adjunction with fluoroscopy and mapping procedures will facilitate treatment of VT and may contribute to the safety of the procedure. PMID- 15113011 TI - Quantitative evaluation of myocardial perfusion in patients with revascularized myocardial infarction: comparison between intravenous myocardial contrast echocardiography and 99mTc-sestamibi single photon emission computed tomography. AB - AIMS: This two-center study compared quantitative segmental perfusion mapping by intravenous myocardial contrast echocardiography (ivMCE) and scintigraphy (SPECT) in patients in the subacute phase of myocardial infarction (AMI). METHODS AND RESULTS: Sixteen patients underwent ivMCE using 1:1 intermittent harmonic imaging 24 h after first AMI treated with PTCA and stenting. Apical contrast echocardiograms were obtained after the injections of Sonazoid. Baseline corrected peak myocardial videointensity (bcPMVI) was determined automatically in 16 segments. Resting 99mTc-sestamibi SPECT was performed within one day after ivMCE. SPECT images were reoriented matching the ivMCE views, and divided into the same segments as in ivMCE, from which mean count rate values were obtained. After exclusion due to artifacts or attenuation, 208/256 (82%) segments remained for analysis. Normalized SPECT count rate and bcPMVI correlated linearly: bcPMVI = 1.237 x SPECT - 35; r = 0.74, p < 0.0001. The relation remained identical in subgroup analysis based on participating center, echocardiographic view, perfusion territory, infarct zone, or function. Using SPECT as reference, mean bcPMVI was 77+/-19% in normal segments, 53+/-29% in mild-moderate defects and 25+/-18% in severe defects (p<0.001 for all comparisons). CONCLUSION: The videointensity increase observed in quantitative ivMCE clearly correlated with SPECT tracer uptake. This further substantiates the use of ivMCE as a valid technique for myocardial perfusion imaging. PMID- 15113012 TI - Transthoracic, harmonic mode, contrast enhanced color Doppler echocardiography in detection of restenosis after percutaneous coronary interventions. Prospective evaluation verified by coronary angiography. AB - AIMS: To test the feasibility and accuracy of transthoracic, harmonic mode, contrast enhanced Doppler echocardiographic assessment of lesion severity after PCI treatment in native coronary arteries--the LAD, Cx and RCA. METHODS AND RESULTS: Prospective evaluation of 59 patients (66 arteries). Restenosis was diagnosed whenever maximal flow velocity at least doubled in comparison to the segment immediately proximal to the PCI site or when local velocity was at least 2 m/s. At 9 months of follow-up final comparison of Doppler echocardiography and coronary angiography was performed with regard to 44 arteries in 40 patients. Of LAD segments assessed, 15 were proximal and 15 middle. The figures for Cx segments were: 4 proximal and 2 mid, and for RCA 2 proximal, 5 middle and 1 distal (i.e. the posterior descending coronary artery). On final coronary angiography there were 8 restenoses--all correctly diagnosed by echocardiography. There were 2 false-positive diagnoses of restenosis. Specificity for restenosis detection was 94% and sensitivity 100%. CONCLUSION: Transthoracic echocardiography allowed for accurate stenosis assessment of principal coronary arteries after successful PTCA. Feasibility of lesion site visualization was 100% for the LAD, 75% for the Cx and 43% for the RCA. Ultrasound contrast agent improved the quality of the RCA images. PMID- 15113013 TI - Churg-Strauss syndrome as an unusual cause of spontaneous atraumatic intra pericardial thrombosis. AB - Intra-pericardial thrombosis is usually due to haemorrhage into the pericardial space or after traumatic pericardiocentesis. An unusual case of spontaneous atraumatic intra-pericardial thrombosis due to Churg-Strauss syndrome, which responded well to immunosuppressive therapy, is presented. PMID- 15113014 TI - Anatomical basis for acquired intracardiac shunt postaortic valve replacement: Doppler echocardiographic diagnosis. AB - We report a case of postoperative intracardiac shunts across the membranous septum detected by Doppler echocardiography and discuss the anatomical basis for the development of such a complication. PMID- 15113015 TI - A rare case of left ventricular outflow obstruction. AB - We report a patient with a mass originating from the anterior mitral valve leaflet causing severe left ventricular outflow tract obstruction. Noninvasive imaging provided the best diagnostic tools for diagnosis. Histological findings showed a very rare giant blood cyst of the mitral valve. PMID- 15113016 TI - Candida sepsis with intramyocardial abscesses mimicking left ventricular noncompaction. AB - Left ventricular (LV) noncompaction is a rare abnormality characterized by more than three trabeculations protruding from the LV wall, distal to the papillary muscles and visible in one echocardiographic image plane. The intertrabecular spaces are perfused from the LV cavity, as visualized on color Doppler imaging. Differential diagnoses of LV noncompaction are intraventricular thrombi, false tendons, aberrant bands, intramyocardial hematoma, cardiac metastases and the apical type of hypertrophic cardiomyopathy. Intramyocardial abscesses have not been reported as a differential diagnosis of LV noncompaction. In the patient presented, cardiac microabscesses due to candida sepsis mimicked LV noncompaction and should be considered in the differential diagnosis of LV noncompaction. PMID- 15113017 TI - Diastolic gradient in hypertrophic cardiomyopathy of the apical type. AB - Diastolic intracavitary gradient in the left ventricle is a common finding in patients with apical hypertrophy. We report the case of a patient with hypertrophic cardiomyopathy and midventricular obstruction. The paradoxical jet flow in this patient, during diastole, was directed towards the base away from the apex. PMID- 15113018 TI - Functional ventricularisation of the left atrium--severe mitral valve prolapse paradoxically resulting in minimal regurgitation. AB - Mitral valve prolapse is a common condition often associated with mitral regurgitation. Intuitively, one would expect a positive correlation between the severity of mitral valve prolapse and the associated regurgitation. This assumption is overturned by an unusual case of functional ventricularisation of the left atrium due to severe mitral valve prolapse which paradoxically resulted in minimal regurgitation. PMID- 15113019 TI - A rapid method to quantify left atrial contractile function: Doppler tissue imaging of the mitral annulus during atrial systole. AB - AIMS: Assess the value of peak atrial systolic mitral annular velocity (Aann) measured by Doppler tissue echocardiography to quantify left atrial systolic function. METHODS: We studied a total of 61 adults; 10 subjects without history of heart disease and 51 patients with a history of atrial fibrillation or undergoing evaluation for left ventricular systolic or diastolic dysfunction. Aann was obtained by averaging peak atrial systolic mitral annular velocities from the septal, lateral, anterior, and inferior annulus. Left atrial fractional area change (FAC) and fractional volume change (FVC) during atrial systole were calculated. The correlation between peak atrial systolic mitral annular velocity (Aann) and left atrial systolic FAC and FVC was determined. RESULTS: Mean FAC and FVC were 27 +/- 12 and 40 +/- 14%, respectively; mean Aann was 11.2 +/- 3.2 cm/s. Linear regression analysis showed correlation between Aann and FAC (r = 0.71; p<0.001) and between Aann and FVC (r = 0.74; p<0.001). CONCLUSIONS: Peak systolic mitral annular velocity correlates well with left atrial systolic FAC and FVC, thus providing an easy means to assess left atrial systolic function. PMID- 15113020 TI - Visualization of a coronary sinus valve using intracardiac echocardiography. AB - Cannulation of the coronary sinus (CS) is sometimes difficult due to the presence of anatomical anomalies. Fluoroscopy is of limited value in visualizing these variations. This case is the first to demonstrate how intracardiac echocardiography (ICE) allows visualization of a valve, which is one of the causes of problematic cannulation of the CS. Based on information obtained by ICE an appropriate catheter could be selected. PMID- 15113021 TI - A versatile synthesis of dihydropyrimidinone C-nucleosides. AB - A versatile synthesis of N-substituted dihydropyrimidinone C-nucleosides (20-29) is described. Glycosyl amino esters (3-9), obtained by reductive alkylation of glycosyl amino esters 1 and 2, on condensation with different isocyanates afforded respective ureido derivatives (10-19) in good to quantitative yields. The latter on cyclative amidation with a combination of DBU/TBAB (tetrabutylammonium bromide)/4A molecular sieve gave the corresponding nucleosides (20-29) in good yields. PMID- 15113022 TI - An unusual "senseless" 2',5'-oligoribonucleotide with potent anti-HIV activity. AB - A new 32-mer 2',5'-oligoribonucleotide of 1-methyl-6-thioinosinic acid (10) has been synthesized. The design of this unique oligoribonucleotide is based on the reported HIV inhibitory activities of both 2',5'-oligonucleotides and the 3',5' oligoribonucleotides containing the 1-methyl-6-mercaptopurine base. Tm and CD studies of 10 revealed that it has no organized secondary structure, presumably due to the rigidity of the molecule. The synthesized oligomer, 10, showed a potent inhibitory effect on HIV-1RF and significantly inhibited HIV-1 reverse transcriptase. PMID- 15113023 TI - Molecular modeling of the binding of 5-substituted 2'-deoxyuridine substrates to thymidine kinase of herpes simplex virus type-1. AB - In an earlier study, De Winter and Herdewijn (J. Med. Chem. 1996, 37, 4727-4737) studied the binding of various 5-substituted 2'-deoxyuridine substrates to thymidine kinase of herpes simplex virus type-1. They used a computational procedure that achieves good correlation with experimentally determined IC50 values. We applied an alternative procedure to the same deoxyuridine substrates, using only three readily calculated quantities-the binding energy, the molecular surface area, and a flexibility factor. Our simplified method achieves the same degree of correlation with the IC50 values as did the earlier procedure. We then applied this procedure to examine the binding of various 5-substituted pyrimidine 1,5-anhydrohexitol substrates to thymidine kinase. PMID- 15113024 TI - Reaction of sugars with 2-hydrazinopyridine, precursors for seco C-nucleosides of 1,2,4-triazolo[4,3-a]pyridine. AB - Reaction of 2-hydrazinopyridine (1) with D-xylose, D-galactose, D-glucose and D fructose afforded the corresponding hydrazones mainly in the acyclic forms 2, 3, 6 and 11 with minor amounts of the cyclic structures. Oxidative cyclization of the hydrazones with bromine in methanol resulted in the formation of the 3 (polyhydroxyalkyl)-1,2,4-triazolo[4,3-a]pyridine derivatives 13-15 whose acetylation afforded the acetylated derivatives 16-18. Assignment of 1D and 2D NMR spectral data in addition to 15N NMR experiments led to complete characterization of the products. PMID- 15113025 TI - Determination of ATP impurity in adenine dinucleotides. AB - Adenine dinucleotides (ApnA) are extracellular signal molecules that are released from blood platelets, following stress, into the vascular system. The most abundant and best-characterized ApnA (Ap4A) interacts with a unique receptor on bovine aortic endothelial cells (BAEC) where it induces nitric oxide. Ap4A also interacts with P2 purinoceptors on BAEC to modulate Ca2+ mobilization and prostacyclin release; this behavior can be equally well explained by Ap4A being either a partial agonist to these receptors, or an antagonist in the presence of ATP contamination. To discern between these two possibilities, we have investigated the presence of such contaminants in ApnA preparations. The studies herein indicate that ApnAs (n = 3-6) contain ATP impurities; thus, when characterizing the ApnA interaction with ATP-binding sites, investigators must assure that the response elicited is not partly due to an ATP impurity. We here provide a means for detecting and estimating ATP impurities within Ap4A preparations while also eliminating them; the level of this contamination is estimated to be as low as 0.2%. We applied our method to distinguish the true effect of Ap4A at P2 purinoceptors; our findings are consistent with Ap4A acting as a partial agonist to these receptors. We also applied our method to characterizing the ApnA interaction with luciferase, and found that decontaminated ApnA (n = 4-6) are weak substrates for luciferase. PMID- 15113026 TI - Acyclonucleoside iron chelators of 1-(2-hydroxyethoxy)methyl-2-alkyl-3-hydroxy-4 pyridinones: potential oral iron chelation therapeutics. AB - The method of synthesizing acyclonucleoside iron chelators is both convenient and cost effective compared to that of synthesizing ribonucleoside iron chelators. The X-ray crystal structural analysis shows that the 2-hydroxyethoxymethyl group does not affect the geometry of the iron chelating sites. Therefore, the iron binding and removal properties of the acyclonucleoside iron chelators should remain similar to the ribonucleoside iron chelators, which is confirmed by the titration and competition reaction of the acyclonucleoside chelators with iron and ferritin, respectively. The acyclonucleoside iron chelators are more lipophilic with measured n-octanol and Tris buffer distribution coefficients than ribonucleoside iron chelators. PMID- 15113027 TI - QSAR analysis for ADA upon interaction with a series of adenine derivatives as inhibitors. AB - The kinetic parameters of adenosine deaminase such as Km and Ki were determined in the absence and presence of adenine derivatives (R1-R24) in sodium phosphate buffer (50 mM; pH 7.5) solution at 27 degrees C. These kinetic parameters were used for QSAR analysis. As such, we found some theoretical descriptors to which the binding affinity of adenosine deaminase (ADA) towards several adenine nucleosides as inhibitors is correlated. QSAR analysis has revealed that binding affinity of the adenine nucleosides upon interaction with ADA depends on the molecular volume, dipole moment of the molecule, electric charge around the N1 atom, and the highest of positive charge for the related molecules. PMID- 15113028 TI - Stereoselective synthesis of novel 4'-branched and bicyclo[3.1.0]hexane-templated nucleoside. AB - The racemic and stereoselective synthesis of a novel nucleoside 4'-branched and bicyclo[3.1.0]hexane templated nucleoside 15 was accomplished using a [3,3] sigmatropic rearrangement, an intramolecular carbene cycloaddition reaction and a Curtius rearrangement as the key reactions. PMID- 15113029 TI - Bis-ketol nucleoside triesters as prodrugs of the antiviral nucleoside triphosphate analogues of 3'-deoxythymidine and 3'-deoxy-2',3' didehydrothymidine. AB - Derivatives of 3'-deoxythymidine (ddT) and 3'-deoxy-2',3'-didehydrothymidine (d4T) were prepared in which the 5'-hydroxyl group of the nucleoside was esterified to a bis-ketol phosphate. The resulting phosphate triesters are postulated to be prodrugs of the corresponding 5'-mononucleotides, which are formed intracellularly by the hydrolysis of the two ketol ester groups. The triesters were tested for anti-HIV activity with the result that those derived from ddT showed enhanced antiviral activity when compared to the parent nucleoside. PMID- 15113030 TI - Expanding the frame: psychoanalysis after September 11. AB - The psychological impact that has emerged following the terrorist attacks on September 11, 2001, raises unique and challenging treatment considerations for the psychoanalyst. The shared trauma of these horrifying events affected many individuals on many levels. This essay considers how analysts will need to demonstrate greater mastery regarding psychological and physiological acute trauma states, further insight into the object-related dynamics of good and evil, and what the analytic frame needs to incorporate to positively influence treatment outcomes. PMID- 15113031 TI - Quality of depressive experiences in borderline personality disorders: differences between patients with borderline personality disorder and patients with higher levels of personality organization. AB - Clinical observations suggest that depressive experiences in patients with borderline personality disorder have a specific quality. These experiences are characterized by emptiness and anger ("angry depression") and are associated with primitive forms of object relations. In this study, this observation was tested empirically. A sample of borderline inpatients (N=30) was compared with a sample of inpatients with higher levels of personality organization suffering from neurotic disorders (N=30). Depression and other affects were assessed by the Affective Dictionary Ulm (Dahl, Holzer, & Berry, 1992). The quality of object relations was assessed by a scale developed by Urist (1977), which was applied to responses in the Holtzman Inkblot Technique (Holtzman, Thorpe, Swartz, & Herron, 1961). Correlations were assessed between depression, on the one hand, and anger, anxiety, and the quality of object relations, on the other hand. The clinical observations were confirmed: In the patients with borderline personality disorder, depression showed significant correlations with the affects of anger, anxiety, and fear, and with primitive forms of object relations. In the patients with higher levels of personality organization, no such correlations were found. The results are discussed with regard to the understanding of borderline disorders, diagnosis, and therapy. PMID- 15113032 TI - Stability of the patient-by-treatment interaction in the Menninger Psychotherapy Research Project. AB - Consistent with the call to consider person-by-treatment interactions in intervention research, Blatt (1992) found that anaclitic and introjective patients responded differently to psychoanalysis and supportive-expressive therapy (SEP) in the Menninger Psychotherapy Research Project (MPRP). Psychoanalysis was significantly more effective than SEP in reducing malevolent, destructive imagery on the Rorschach among introjective patients, those patients who are primarily preoccupied with control and self-definition. Conversely, SEP was significantly more effective than psychoanalysis in reducing these malevolent, destructive images among anaclitic patients, those patients who are primarily preoccupied with interpersonal relatedness. The present analyses of data from the MPRP demonstrate the stability of this statistically significant patient-by-treatment interaction even in the subsample of patients for whom the anaclitic-introjective distinction was ambiguous, reaffirming the validity of both the anaclitic-introjective distinction and the importance of considering patient characteristics in psychotherapy research and practice. PMID- 15113033 TI - Physicians' wives evaluate their marriages, their husbands, and life in medicine: results of the AMA-Alliance Medical Marriage Survey. AB - This largest-ever investigation of the attitudes, lifestyles, and marital adjustment of physicians' wives disputes many stereotypes of medical marriage. Responses of 603 members of the American Medical Association Alliance to standardized and subjective measures indicated high levels of marital adjustment and overall satisfaction with work/life balance. Wives' marital adjustment was affected by age of oldest child, husbands' work hours, and wives' work outside the home. It is proposed that, more than hours worked, it is how a couple treats each other when they are not working that most powerfully determines the quality of a contemporary medical marriage. Wives' advice on marital and work/life issues is summarized. PMID- 15113034 TI - Profiles of impaired health professionals. AB - There are numerous studies that describe the characteristics of impaired health professionals and the types of professional misconduct leading to licensing board action. These studies have two fundamental limitations. The first is the sampling procedure, and the second is that they typically do not examine health professionals who are currently in treatment. This study describes the problems that lead health professionals--comprising psychiatrists, nonpsychiatric physicians, psychologists and social workers--to seek treatment and the sources of referral for treatment. A total of 334 health professionals were studied who sought out an evaluation or treatment at The Menninger Clinic between 1985 and 2000. The findings indicated that the participants' therapist was the largest referral source and that the most commonly cited problems leading to referral were marital and emotional difficulties rather than substance abuse, boundary violations, or prescribing problems. Licensing and regulatory agencies can take proactive steps to identify professionals with social and emotional vulnerabilities who may be at greater risk for unethical and negligent behavior. PMID- 15113035 TI - IgM serum antibodies to Epstein-Barr virus are uniquely present in a subset of patients with the chronic fatigue syndrome. AB - BACKGROUND: A unique subset of patients with chronic fatigue syndrome (CFS) and IgM serum antibodies to cytomegalovirus (HCMV) non-structural gene products p52 and CM2 (UL 44 and UL 57) has been described. PATIENTS AND METHODS: Fifty-eight CFS patients and 68 non-CFS matched controls were studied. Serum antibodies to EBV viral capsid antigen (VCA) IgM and EBV Early Antigen, diffuse (EA, D) as well HVCMV(V), IgM and IgG; VP (sucrose, density purified V); p52 and CM2 IgM serum antibodies were assayed. RESULTS: Mean age of CFS patients was 44 years (75% women). Control patients were 9 years older (73% women). Serum EBV VCA IgM positive antibody titers were identified in 33 CFS patients (Group A subset EBV VCA IgM 62.3+/-8.3, neg. <20), but were not present in other CFS patients, (Group B subset EBV VCA IgM 6.8+/-0.7) controls (p<0.0001). EBV VCA IgM titers remained positive in CFS patients from Group A for 24-42 months. CONCLUSION: Serum antibody to EBV VCA IgM may be a specific diagnostic test for a second subset of CFS patients. PMID- 15113036 TI - Changes of amino acid serum levels in pediatric patients with higher-risk acute lymphoblastic leukemia (CCG-1961). AB - BACKGROUND: Deamination of asparagine (Asn) and glutamine (Gln) by asparaginases (ASNase) is associated with good prognosis in acute lymphoblastic leukemia (ALL). Chemotherapy drugs used for ALL may accelerate catabolism of other amino acids (AA). MATERIALS AND METHODS: We studied ASNase activity and changes of Asn, Gln, serine (Ser), threonine (Thr), histidine (His), proline (Pro) and arginine (Arg) levels and sought relationships in sera from 73 pediatric ALL patients, who received ASNase-containing chemotherapy. RESULTS: Asparaginase activity averaged 0.4+/-0.34 IU/ml (mean+/-SDEV) in all specimens. All AA decreased after treatment, ranging from 18.6%-82.6% of control. Asparaginase activity of 0.7 IU/ml provided 90% Asn and Gl deamination. The data were dichotomized in subsets of low ASNase (range 0.02-0.39 IU/ml, mean=0.17+/-0.09 IU/ml) and high ASNase (range 0.4-1.69 IU/ml and mean=0.72+/-0.32 IU/ml). Asparagine and Gln % deamination values were correlated with ASNase activity (p=0.0002 and p=0.0001). Similarly, decreases of Arg and Ser levels were also correlated, p =0.0009 and p=0.032, respectively. In the high ASNase subset, a 39% decrease of Arg and 26% of Ser was obtained. Low ASNase activity was correlated with lower Asn and Gln % deamination and with moderate decrease of Ser (14.6%) and Arg (19.6%). Threonine, Pro and His also decreased, but no correlations were obtained with ASNase activity. CONCLUSION: Asparagine, Gln and five other AA declined during ASNase treatment. Asparagine and Gln % deamination values are highly correlated with serum ASNase activity. Asparaginase may indirectly cause moderate depletion of serum Arg and Ser levels, providing an enhancement in leukemia blasts apoptosis. Toxicity from the ASNase and other drugs could enhance the decrease of AA serum levels. Further studies are needed to verify these findings and their potential clinical importance in the treatment of ALL patients. PMID- 15113037 TI - Antisense therapy specific to mutated K-ras gene in hamster pancreatic cancer model. Can it inhibit the growth of 5-FU and MMC-resistant metastatic and remetastatic cell lines? AB - K-ras point mutation at codon 12 has a relationship greater than 90% with pancreatic cancer. Cancer therapy should also include the treatment of metastatic disease because it is known that the properties of metastatic cells may vary considerably from those of the primary tumor. AIM: To clarify if the same drugs, which can inhibit the tumor growth in the parental cell line, can inhibit the pancreatic metastatic and remetastatic cell lines at the same concentrations and to compare the inhibition with antisense oligonucleotides mismatched to K-ras gene, in Syrian golden hamsters. MATERIALS AND METHODS: HaP-T1, a BHP-induced hamster pancreatic cancer cell line, MS-PaS-1 (a metastatic cell line established from "return trip" metastases from the liver to the pancreas) and MS-PaS-2 named as a "remetastatic cell line", i.e., metastases from MS-PaS-1 were used. MTT and MTT-agarose assays were performed, using 5-Fluorouracil (5-FU), Mitomycin C (MMC) and antisense oligonucleotide specific to K-ras oncogene. RESULTS: The inhibitory concentration (IC50) of 5-FU, which inhibited HaP-T1, had to be increased by 50 fold to inhibit MS-PaS-1 and 100-fold to inhibit MS-PaS-2. MMC had to be increased by 10-fold to inhibit MS-PaS-1 and 50-fold to inhibit MS-PaS-2. However, IC50 was the same when antisense oligonucleotide was tried in these 3 cell lines. CONCLUSION: Antisense oligonucleotide-targeted K-ras gene may be a good choice for therapy because it could inhibit the growth in metastatic and remetastatic cells as well as in primary tumor cells. PMID- 15113038 TI - Analgesic effect of intrathecal administration of orexin on neuropathic pain in rats. AB - Orexin-A, a hypothalamic peptide found in the neurons of the lateral hypothalamus, has been shown to modulate pain. We examined whether orexin could alleviate heat-evoked hyperalgesia in rats caused by chronic constriction injury (CCI) of the sciatic nerve. Orexin-A, orexin-B, the vehicle, or orexin-A antiserum was intrathecally administered to CCI rats. Paw withdrawal latency (PWL) was measured from 30 to 300 minutes after injection, which was repeated for 2 days. Orexin-A administration normalized deltaPWL (PWL in the CCI side minus PWL in the control side) and inhibited heat-evoked hyperalgesia in CCI rats, while orexin-A antiserum inhibited the normalization of heat-evoked hyperalgesia caused by orexin-A two-fold. In contrast, orexin-B had no significant effect. These results suggest that orexin-A may be applicable for treatment of neuropathic pain. PMID- 15113039 TI - Acute effects of low-frequency electromagnetic fields on leukocyte-endothelial interactions in vivo. AB - BACKGROUND: To clarify the effect of extremely low frequency electromagnetic field (ELF-EMF) on the microcirculatory system, acute effects on leukocyte endothelium interactions resulting from ELF-EMF exposure were examined with conscious BALB/c mice by means of a dorsal skinfold chamber. MATERIALS AND METHODS: The fluorescent dye, rhodamine 6G, was injected into the caudal vein to visualize leukocytes in vivo and to analyze leukocyte-endothelium interactions. Mice were exposed in vivo to 50 Hz linear sinusoidal wave EMF at magnetic flux densities of 3, 10 or 30 mT, for 30 minutes. RESULTS: The mean velocity of free flowing leukocytes in the center stream of venules revealed no distinct changes following exposure. However, at a flux density of 30 mT, the number of adherent leukocytes, defined as the total number of rolling and endothelium-adhering leukocytes, increased significantly following ELF-EMF exposure (p<0.05); no significant differences in adherent cell numbers were found in the 3 mT and 10 mT exposure groups or sham controls comparing pre- and post-exposure periods. CONCLUSION: These results demonstrated that high magnetic flux densities of ELF EMF had an effect on leukocyte-endothelium interactions and indicated that a threshold level for this phenomenon exists in the range of 10-30 mT under these experimental conditions. PMID- 15113040 TI - Expression of multi-drug resistance genes (mdr1, mrp1, bcrp) in primary oral squamous cell carcinoma. AB - The expression of resistance genes can cause the ineffectiveness of chemotherapeutics for the treatment of cancer. Therefore, known resistance genes were investigated in oral squamous cell carcinoma (OSCC) and the results were compared with clinico-pathological findings. MATERIALS AND METHODS: Fresh frozen samples of 45 primary OSCC were investigated for the expression of mdr1 (p glycoprotein-mediated multi-drug resistance), mrp1 (multi-drug resistance-related protein) and bcrp (breast cancer-related protein), using a reverse transcriptase PCR. The gene products were revealed immunohistochemically on representative slices of the same tumor sample. The results were compared with TNM stage grouping [SG, (UICC, 1987)], HPV infection and p53 mutations (exons 5-8). RESULTS: The expression of the resistance genes was independent of age, sex, localisation of the tumor, HPV infection and p53 mutations. SG did not correlate to mdr1 and mrp1. On the other hand, bcrp expression increased 2.7-fold between SG III and IV OSCC. Loss of differentiation was associated with an increased expression of mdr1 (p=0.06), mrp1 (p<0.01) and bcrp (p<0.01). The bcrp expression correlated with shorter survival periods. Expression of mrp1 and mdr1 did not correlate positively in a linear pattern. Expression of mdr1 and bcrp moderately positively correlated (p<0.01). DISCUSSION: Multi-drug resistance genes can be up regulated in OSCC. The expression of at least one of these genes is up-regulated in SG-IV OSCC. Determining these genes could probably support current studies on therapeutic effects in OSCC, e.g. new cytostatic drugs. PMID- 15113041 TI - Percutaneous kyphoplasty: new treatment for painful vertebral body fractures. AB - AIMS AND BACKGROUND: The purpose of this study was to assess the effectiveness and safety of Percutaneous Kyphoplasty as a new method of treatment for pain deriving from vertebral compression fractures (VCF). PATIENTS AND METHODS: We treated sixteen patients with unremitting pain over spine, which increased particularly when pressure was applied over the spinous process, in absence of neurological signs and refractory to conventional medical therapy. RESULTS: The method demonstrated swift pain relief associated with an evident augmentation in the resistance and restoration of the vertebral body's physiological shape. Polymethylmethacrylate (PMMA) leakages were not observed in the epidural space or foraminal area. The presence of complications such as pulmonary embolism involving the venous plexus, toxicity due to PMMA and infection due the procedure did not occur. CONCLUSION: Kyphoplasty is an effective, alternative, simple and safe treatment of vertebral collapse consequent to osteoporosis. aggressive haemangiomas, myelomas and metastases. PMID- 15113042 TI - Expression of vascular endothelial growth factor (VEGF) and association with microvessel density in benign prostatic hyperplasia and prostate cancer. AB - BACKGROUND: Tumor angiogenesis is an absolute requirement for tumor growth and a prognostic factor for various malignant neoplasms. Recent reports in the literature have addressed the importance of the VEGF system in benign prostatic hyperplasia (BPH) and adenocarcinoma, however the results are controversial. The aim of the present study was to determine and compare the levels of VEGF expression and vascularity in BPH and prostate carcinoma. MATERIALS AND METHODS: We examined 60 prostate adenocarcinomas and 64 benign prostatic hyperplasias. Angiogenesis was estimated by determining microvessel counts (MVC), with the use of anti-CD31 and anti-CD34 antibodies. Expression of VEGF was also evaluated immunohistochemically. RESULTS AND CONCLUSION: Our data showed that angiogenesis was more prominent in carcinomas than in BPH. Furthermore, increased MVC was significantly associated with high-grade carcinomas. Angiogenesis was correlated with VEGF expression and it was, at least in part, mediated by the latter. Thus, prostate adenocarcinoma may represent a suitable neoplasm for antiangiogenic treatment in combination with conventional therapies. PMID- 15113043 TI - A novel antioxidant non-steroidal anti-inflammatory agent protects rat liver against ischemia-reperfusion injury. AB - Liver ischemia followed by reperfusion is an important and common clinical event. A major mechanism is leukocyte adhesion to endothelium followed by release of reactive oxygen metabolites. The aim of this study was to determine the effects of a novel antioxidant ethylenediamine derivative with anti-inflammatory properties (compound IA) on an imitated clinical setting of acute hepatic ischemia-reperfusion injury. Eight groups of rats were subjected to a model of hepatic ischemia that was produced by occluding for 30 min the portal vein and hepatic artery. At the end of ischemia, compound IA was administered intravenously and the clamps were removed allowing reperfusion for 60 min or 24 h. The effect of compound IA was evaluated by histopathological examination, lipid peroxidation and plasma levels of liver enzymes. Administration of compound IA resulted in significantly less histological damage in liver tissue after 30 min ischemia followed by 60-min and 24-h reperfusion. Ischemia followed by 60 min of reperfusion increased lipid peroxidation compared to the sham-operated and the non-ischemic group. This increase was attenuated in the group treated with compound IA. Serum enzyme levels were significantly higher in the reperfusion groups compared to the non-ischemic groups and diminished after treatment. Compound IA exerted a protective effect on hepatic reperfusion injury in rats. Compound IA is believed to act by means of its potent antioxidant and anti inflammatory activities. PMID- 15113044 TI - Cytotoxicity, cytoprotection and neurotoxicity of novel deprenyl-related propargylamines, stable nitroxide free radicals, in vitro and in vivo. AB - Since novel synthesized deprenyl-related derivatives of nitroxides, named "JSAKs", have been shown to possess antioxidative properties, their cytotoxicity on neuronal-like PC-12 cells line was examined. The antiproliferative effect of two selected JSAKs was examined and expressed as IC10, IC50 and IC90, and compared with those of the parent nitroxide (Nx-640), model nitroxide TEMPO and deprenyl. There were substantial differences in the dose-dependence of all the observed antiproliferative and cytotoxic effects. Compared to anticancer drugs and apoptosis inducers with topoisomerase inhibitor properties (etoposide and camptothecin), novel compounds displayed cytotoxicity at considerably higher concentrations. The dose-dependent anti-apoptotic potency of JSAKs and Nx-640 was also investigated and compared to TEMPO and deprenyl effects. The observed structure-dependent correlation was very encouraging and prompted us to screen and to compare the in vivo time-dependent effects of JSAKs, Nx-640 and deprenyl administration on the rat intact nigrostriatal neurocytes. TH-immunochemistry was applied as the test method and marker for the changes in the state of the rat catecholaminergic system, also giving evidence that low-toxic and cell-permeable JSAKs can cross the blood-brain barrier, which is the mandatory prerequisite for the therapeutic application of antioxidants and drugs to the brain. Taken together, it can be concluded with great certainty that novel deprenyl-related JSAKs might be especially good candidates for further anticancer investigations in vitro and in vivo and future pharmacological applications. PMID- 15113045 TI - Biodistribution of injected tritiated hyaluronic acid in mice: a comparison between macromolecules and hyaluronic acid-derived oligosaccharides. AB - BACKGROUND: Hyaluronan (HA) has been reported to bind specifically and with high affinity to various cell types and to directly modify cell behaviour. In a previous report we demonstrated that both high molecular weight molecules (HA(H)) and HA-derived oligosaccharides were efficient at triggering terminal differentiation of acute myeloid leukemia (AML) blasts, in vitro, through CD44 ligation. MATERIALS AND METHODS: To explore the possibility of using HA for a differentiation therapy in AML, we investigated whether intravenous injection of tritiated HA(H) and/or HA-derived oligosaccharides (HA10-20) into mice accumulated in bone marrow, the main site of AML cell proliferation. RESULTS: The present work showed that the level of HA in bone marrow: 1) was maximum 5 hours after injection of either HA(H) or HA10-20; 2) was about 40 times higher after HA(H) than after HA10-20 injection. The amount of HA in bone marrow (5.8% ID/g) was two-fold higher than in serum, indicating that it was not due to circulating blood. Finally, using chromatographic analysis, we showed that about 34% of tritiated HA present in bone marrow 5 hours after HA(H) injection displayed a size higher or equal to HA10. CONCLUSION: After a single injection of macromolecular hyaluronan in mouse bone marrow we obtained a concentration of oligosaccharides close to the one shown to trigger AML cell differentiation in vitro. A part of the oligosaccharides had a size higher than or equal to the minimal one required to interact with HA receptors. PMID- 15113047 TI - The chronological appearance of flat colonic neoplasias in rats. AB - Following weekly s.c. injections of the colonotropic carcinogen 1,2 dimethylhydrazine (DMH), we investigated the occurrence of flat and protruding neoplasias in the colon of rats at various time intervals. Forty-seven DMH treated rats were sacrificed at the 13th, 15th, 19th, 21st and 22nd weeks. A total of 88 tumors evolved in 35 of the 47 DMH-treated rats. The number of neoplasias/animal was 0.3 at week 13, 1.2 at week 15, 2.0 at week 19, 2.5 at week 21 and 4.0 at week 22. In the right colon, although the percent of flat adenomas was lower than of protruding adenomas, the percent of flat carcinomas was significantly higher than of protruding carcinomas, indicating that flat adenomas progress more rapidly to invasive carcinoma than protruding adenomas in the right colon. The opposite was recorded in the left colon where the percent of protruding adenomas was lower than of flat adenomas, but the percent of protruding carcinomas was higher than of flat carcinomas. During the last experimental week as many as 63% of the protruding carcinomas occurred but only 25% of the flat carcinomas. These experimental results seem to substantiate previous observations in humans suggesting that, in the colonic mucosa, flat and protruding adenomas follow different pathways of neoplastic transformation. PMID- 15113046 TI - Role of p53, CD44V6 and CD57 in differentiating between benign and malignant follicular neoplasms of the thyroid. AB - The distinction between follicular adenoma (FAD) and follicular carcinoma (FCA) of the thyroid can be particularly challenging in routine practice of diagnostic surgical pathology. It often requires examination of several histologic sections in order to identify the presence of unequivocal capsular and/or vascular invasion. To investigate the role of immunohistochemical markers in the differential diagnosis of follicular lesions of the thyroid, we studied the pattern of expression of p53, Bcl-2 and of the adhesion molecules CD44V6 and CD57, in 20 FADs and 21 FCAs of the thyroid. Ninety percent of FCAs exhibited strong nuclear p53 expression. p53 stain was seen in only 15% of FADs (p<0.0001), and it was weak. Bcl-2 cytoplasmic immunoreactivity was observed in 57% of FCAs and 60% of FADs (p=1.00). Similarly, membranous CD44V6 staining was seen in 81% of FCAs, but in only 20% of FADs (p=0.0001). CD57 was present in the cytoplasm of 71% of FCAs and 15% of FADs (p=0.0004). None of the markers studied correlated with tumor size. The results of this study indicate that immunohistochemical detection of p53, CD44V6 and CD57 may have practical utility in the differential diagnosis of FCAs from FADs in routine surgical pathology. PMID- 15113048 TI - Involvement of IL-6 and IL-6 receptor in fibrinogen synthesis in the liver of Triton WR-1339-induced hyperlipidemic rats. AB - To clarify the mechanisms by which Triton WR-1339 causes an elevation in the plasma fibrinogen level, we studied the time courses of hepatic mRNA expression for beta-chain fibrinogen, interleukin-6 (IL-6) and IL-6 receptor (R) by RT-PCR. After intravenous injection of Triton WR-1339 (150, 300 and 500 mg/kg) in Sprague Dawley rats, the plasma level of fibrinogen and MCP-1 significantly and dose relatedly increased from 12 to 24 hours (h). At 3 and 6 hours dose dependent increases were found in hepatic IL-6 and IL-6R mRNA expression with increases in the hepatic mRNA expression for beta-chain fibrinogen as the rate-limiting step in fibrinogen synthesis. These results suggest that the increase in the plasma fibrinogen level is followed by the enhancement of hepatic mRNA expression of beta-chain fibrinogen, IL-6 and IL-6R, that is, IL-6 and IL-6R may partly regulate the plasma fibrinogen level in Triton WR-1339-induced hyperlipidemic rats. PMID- 15113049 TI - DNA-flow cytometry, DNA-image cytometry and proliferation index (MIB-5) in irradiated rat salivary glands and salivary gland tumors. AB - AIM: Salivary glands (SG) can become atrophic following radiation exposure. Malignant transformation of SG in a radiation field is another known sequela of patients who have been treated by radiotherapy for a malignant tumor in the head and neck region. The aim of this study was to investigate cytogenetic alterations and to determine the proliferation index (PI) of SG of rats subjected to various total dosages of fractionated X-rays. MATERIALS AND METHODS: We investigated rat SG, subjected to 20, 40, or 60 Gy exposure by X-rays to the left neck and skull base. Non-irradiated rats served as a control group. Tumors originating from the SG were histologically-diagnosed following the descriptions for human SG tumors. The MIB-5 antibody was used to determine the PI. The ploidy was determined by flow and image cytometry (FCM, ICM). RESULTS: We consistently recorded diploid histograms in the FCM in irradiated glands. ICM revealed aneuploid histograms in 6/22 tumors, 3 of them were Auer Type III or IV. The PI showed a dose- and time dependent course, indicative of variable regeneration properties of the parenchyma. Statistically significant differences were found for the PI within the irradiation groups and comparing irradiated SG and tumors. CONCLUSION: Irradiation of rat SG can cause almost complete loss of function. On the other hand, the PI remained in animals subjected to 40 Gy and investigated 1 year after completion of radiation at a level up to 10-fold higher than in untreated controls. The PI in carcinoma is higher in this species than in irradiated SG. Constantly elevated PI could support the development of cancer in SG. PMID- 15113050 TI - Erianin induces a JNK/SAPK-dependent metabolic inhibition in human umbilical vein endothelial cells. AB - BACKGROUND: Erianin is a natural product derived from Dendrobium chrysotoxum, with promising antitumor activity. MATERIALS AND METHODS: To evaluate the metabolic effect of erianin, a cytosensor assay for acidification rate, MTT assay, measurement of lactate, glucose and ATP were performed in human umbilical vein endothelial cells (HUVECs) exposed to 1-100 nM erianin. JNK/SAPK activity was detected by Western blot. RESULTS: Twelve- or 24- hour incubation with erianin induced a dose-dependent metabolic inhibition, as indicated by reduced acidification rate and cell viability, with an endothelium-selectivity. Erianin caused decreases in lactate production, glucose consumption and intracellular ATP level. Pretreatment with the JNK/SAPK inhibitor SP600125 significantly abolished these inhibitory responses, and especially restored the erianin-induced decreases in ATP and the erianin-induced phosphorylation of JNK/SAPK with dose- and time- dependence. CONCLUSION: Erianin inhibited endothelial metabolism in a JNK/SAPK dependent manner. This mechanism may be involved in the potential antitumnor and antiangiogenic actions of erianin. PMID- 15113051 TI - Non-invasive diagnostic and functional evaluation of cardiac and pulmonary involvement in systemic sclerosis. AB - BACKGROUND: The aim of the present study was to evaluate the role of non-invasive methods in the early detection of pulmonary and cardiac involvement in Systemic sclerosis (SSc) and to identify clinical and/or instrumental patterns of prognostic value. PATIENTS AND METHODS: Twenty female patients affected by SSc (8 with diffuse cutaneous SSc and 12 with limited cutaneous SSc) were enrolled in our study. Cardiac and pulmonary involvement (respiratory function tests and carbon monoxide lung diffusion [DLCO], chest radiography, high resolution computed tomography [HRCT] and lung perfusion magnetic resonance) were evaluated. RESULTS: All 18 patients studied with respiratory function tests showed a significant reduction of DLCO. HRCT was considerably more sensitive than traditional chest radiography (59% versus 28%; p<0.05). Lung perfusion MRI revealed normal findings in 15 patients. Abnormal lung perfusion MRI results were found only in 3 patients. Angina pectoris with electrocardiographic and scintigraphic ischemic changes, severe regional wall motion abnormalities and complex arrhythmias seemed to be associated with poor prognosis. CONCLUSION: Taken together these results indicate that a pulmonary involvement occurs both in limited and in diffuse cutaneous SSc patients and develops, in 83% of the cases, without any regional lung perfusion abnormality. Furthermore, cardiac involvement is detected in 65% of the cases as a consequence of a range of noxious events including myocardial ischemia, fibrosis and pressure overload which may result in ventricular dysfunction and arrhythmias. Lung perfusion MRI should be considered as a complementary diagnostic method for the functional evaluation of these symptoms in systemic sclerosis. PMID- 15113052 TI - Modulation of multidrug resistance and apoptosis of cancer cells by selected carotenoids. AB - The multidrug resistance (MDR) proteins that belong to the ATP-binding casette superfamily are present in a majority of human tumors and are an important final cause of therapeutic failure. Therefore, compounds which inhibit the function of the MDR-efflux proteins may improve the cytotoxic action of anticancer chemotherapy. The effects of carotenoids were studied on the activity of the MDR 1 gene-encoded efflux pump system. The carotenoids, isolated from paprika and other vegetables, were tested on the rhodamine 123 accumulation of human MDR-1 gene-transfected L1210 mouse lymphoma cells and human breast cancer cells MDA-MB 231 (HTB-26). Capsanthin and capsorubin enhanced the rhodamine 123 accumulation 30-fold relative to nontreated lymphoma cells. Lycopene, lutein, antheraxanthin and violaxanthin had moderate effects, while alfa- and beta-carotene had no effect on the reversal of MDR in the tumor cells. Apoptosis was induced in human MDR1 transfected mouse lymphoma cells and human breast cancer MDA-MB-231 (HTB-26) cell lines in the presence of lycopene, zeaxanthin and capsanthin. The data suggest the potential of carotenoids as possible resistance modifiers in cancer chemotherapy. PMID- 15113053 TI - Ethical principles in end-of-life decisions in different European countries. AB - The majority of ICU deaths are preceded by a decision to limit treatment in some way. Decisions to withhold or withdraw treatment vary considerably depending on many factors including local practice, cultural and religious background, family and peer pressure. Here we will discuss the current situation across Europe, based on the findings from three large international studies. PMID- 15113054 TI - Does mechanical bowel preparation have a role in preventing postoperative complications in elective colorectal surgery? AB - BACKGROUND: mechanical bowel preparation (MBP) consists of orthograde fluid preparation to clean the bowel. MBP is considered to prevent postoperative complications. METHODS: meta-analysis of prospective randomised clinical trials (RCT) evaluating MBP versus no MBP. RESULTS: following a medline search we retrieved 15 prospective trials of which only 5 where RCT comparing MBP versus no MBP in elective colorectal surgery. For the randomised studies, there were respectively 186, 179, 149, 267 and 380 patients, including all type of resections. The total number of patients in these 5 studies was 1144 (565 with MBP and 579 with no preparation). All patients received perioperative antibiotic prophylaxis. Only one of these RCT show a significant decrease in anastomotic leak (AL), but among all the patients enrolled, AL is significantly more frequent in the group with MBP (Odds Ratio 1.8). Wound infection, re-operation and intra abdominal abscess rates were more frequent in the MBP group but the difference did not reach statistical significance and the odds ratios for a 95% confidence interval were extremely large. CONCLUSION: there is limited evidence in the literature to support the use of MBP in patient undergoing elective colorectal surgery. Available data tend to suggest that MBP could be harmful with respect to the incidence of anastomotic leak. Moreover, MBP does not reduce the incidence of other infectious complications. Further RCTs are needed to establish an evidence based rationale for the use of MBP in elective colorectal surgery. PMID- 15113055 TI - Thoracic aortic plaques, transoesophageal echocardiography and coronary artery disease. AB - BACKGROUND: The purpose of this study was to assess whether the detection of atherosclerotic aortic plaques by transoesophageal echocardiography (TEE) could be used as a marker of coronary artery disease (CAD), relying on their number, cross-sectional surface, depth and localisation. METHODS: The thoracic aortas of 102 consecutive patients (77 men, mean age 67 +/- 12 years) undergoing elective cardiac surgery were assessed by TEE. Atherosclerotic plaques were defined as > or = 5 mm thick focal hyperechogenic zones of the aortic intima and/or lumen irregularities with mobile structures or ulcerations. All patients had undergone prior coronary angiography. RESULTS: Thoracic aortic plaques were present in 73 patients, 66 of whom had CAD. The presence of aortic plaques detected by TEE identified significant coronary artery disease with a sensitivity of 90% and a specificity of 76%. The maximum transverse cross-sectional plaque area, the maximum plaque depth and the total plaque number all correlated significantly with the presence of CAD, but not with its severity. Multivariate regression analysis showed that aortic plaques, hypertension and hypercholesterolaemia were significant predictors of CAD, but aortic plaques were the most significant predictor regardless of age and sex. CONCLUSIONS: This study suggests that detection of atherosclerotic aortic plaques is a useful marker of significant coronary artery disease. Absence of plaques in the patients aged over 70 identified a subgroup with a very low probability of CAD. PMID- 15113056 TI - Delayed-onset and long-lasting severe neutropenia due to rituximab. AB - BACKGROUND: Rituximab, a monoclonal antibody, is effective in CD20-positive B cell lymphoma and is now widely used either as a single agent or in combination with chemotherapy. The antibody's toxicity is generally mild and transient. There are reports of protracted neutropenia in patients treated with rituximab. CASE REPORT: We report on a patient with Burkitt's lymphoma treated with the hyper CVAD chemotherapy regimen combined with rituximab. Four weeks after the five months' treatment marked neutropenia and hypogammaglobulinaemia occurred and persisted for one year. Both laboratory findings were not associated with severe infections in our patient. CONCLUSIONS: Delayed-onset neutropenia is a newly recognised toxicity of rituximab treatment which may last up to one year and be complicated by serious infections. PMID- 15113057 TI - State of the art in diagnostic liver imaging. PMID- 15113058 TI - Advantages and differences of Resovist towards Gadolinium contrast in MRI with focus on metastatic liver lesions. PMID- 15113059 TI - Resovist for imaging of hepatocellular carcinoma in the cirrhotic liver. PMID- 15113060 TI - Experience with Resovist in gastroenterologic radiology and intervention. PMID- 15113061 TI - Clinical impact of Resovist-enhanced MRI: results of an Italian multicenter trial. PMID- 15113062 TI - Preoperative patient work-up: Resovist-enhanced MRI vs. multislice CT- preliminary results of an ongoing study. PMID- 15113063 TI - Health economic evaluation of liver MRI in colorectal cancer patients. PMID- 15113064 TI - Quo vadis diagnostic liver imaging? PMID- 15113065 TI - The utility of structure-activity relationship (SAR) models for prediction and covariate selection in developmental toxicity: comparative analysis of logistic regression and decision tree models. AB - Structure-activity relationship (SAR) models can be used to predict the biological activity of potential developmental toxicants whose adverse effects include death, structural abnormalities, altered growth and functional deficiencies in the developing organism. Physico-chemical descriptors of spatial, electronic and lipophilic properties were used to derive SAR models by two modeling approaches, logistic regression and Classification and Regression Tree (CART), using a new developmental database of 293 chemicals (FDA/TERIS). Both single models and ensembles of models (termed bagging) were derived to predict toxicity. Assessment of the empirical distributions of the prediction measures was performed by repeated random partitioning of the data set. Results showed that both the decision tree and logistic regression derived developmental SAR models exhibited modest prediction accuracy. Bagging tended to enhance the prediction accuracy and reduced the variability of prediction measures compared to the single model for CART-based models but not consistently for logistic-based models. Prediction accuracy of single logistic-based models was higher than single CART-based models but bagged CART-based models were more predictive. Descriptor selection in SAR for the understanding of the developmental mechanism was highly dependent on the modeling approach. Although prediction accuracy was similar in the two modeling approaches, there was inconsistency in the model descriptors. PMID- 15113066 TI - Performance of (consensus) kNN QSAR for predicting estrogenic activity in a large diverse set of organic compounds. AB - A novel method (in the context of quantitative structure-activity relationship (QSAR)) based on the k nearest neighbour (kNN) principle, has recently been introduced for the derivation of predictive structure-activity relationships. Its performance has been tested for estimating the estrogen binding affinity of a diverse set of 142 organic molecules. Highly predictive models have been obtained. Moreover, it has been demonstrated that consensus-type kNN QSAR models, derived from the arithmetic mean of individual QSAR models were statistically robust and provided more accurate predictions than the great majority of the individual QSAR models. Finally, the consensus QSAR method was tested with 3D QSAR and log P data from a widely used steroid benchmark data set. PMID- 15113067 TI - Kinetics of the radical scavenging activity of beta-carotene-related compounds. AB - To clarify the non-enzymatic radical-scavenging activity of beta-carotene-related compounds and other polyenes, we used differential scanning calorimetry to study the kinetics of radical polymerization of methyl methacrylate (MMA) by 2,2' azobisisobutyronitrile (AIBN) or benzoyl peroxide (BPO) in the absence or presence of polyenes under nearly anaerobic conditions at 70 degrees C, and analyzed the results with an SAR approach. The polyenes studied were all-trans retinol, retinol palmitate, calciferol, beta-carotene and lycopene. Polyenes produced a small induction period. The stoichiometric factor (n) (i.e. the number of radicals trapped by each inhibitor molecule) of polyenes was close to 0. Tetraterpenes (beta-carotene, lycopene) suppressed significantly more of the initial rate of polymerization (R(inh)) than did diterpenes (retinol, retinol palmitate). The inhibition rate constants (k(inh)) for the reaction of beta carotene with AIBN- or BPO-derived radicals were determined to be 1.2-1.6x10(5) l/mol s, similar to published values. A linear relationship between (k(inh)) and the kinetic chain length (KCL) for polyenes was observed; as (k(inh)) increased, KCL decreased. KCL also decreased significantly as the number of conjugated double bonds in the polyenes increased. Polyenes, particularly beta-carotene and lycopene, acted as interceptors of growing poly-MMA radicals. PMID- 15113068 TI - e-Quantum chemistry free resources. AB - Among computational chemistry methods, quantum mechanics calculates geometries and electronic structures with accuracy especially for systems with electronic delocalization. The use of a multiconfigurational approach is able to treat highly degenerated states such as those occurring at the transition state in some chemical reactions. Moreover, an accurate description of potential energy surfaces can be obtained with the evaluation of the dynamic electron correlation effects by this approach. Molecular properties range from simple dipole moments, vibrational frequencies or IR intensities to frequency dependent hyperpolarizabilities. Quantum chemical calculations are thus an attractive source of molecular descriptors which can be used in QSAR/QSPR studies and which can express all electronic and geometric properties of molecules. A survey and a comparison of the performance of free e-resources for semi-empirical and ab initio calculations is provided. PMID- 15113069 TI - Structure-activity approach to the identification of environmental estrogens: the MCASE approach. AB - A sizable number of environmental contaminants and natural products have been found to possess hormonal activity and have been termed endocrine-disrupting chemicals. Due to the vast number (estimated at about 58,000) of environmental contaminants, their potential to adversely affect the endocrine system, and the paucity of health effects data associated with them, the U.S. Congress was led to mandate testing of these compounds for endocrine-disrupting ability. Here we provide evidence that a computational structure-activity relationship (SAR) approach has the potential to rapidly and cost effectively screen and prioritize these compounds for further testing. Our models were based on data for 122 compounds assayed for estrogenicity in the ESCREEN assay. We produced two models, one for relative proliferative effect (RPE) and one for relative proliferative potency (RPP) for chemicals as compared to the effects and potency of 17beta estradiol. The RPE and RPP models achieved an 88 and 72% accurate prediction rate, respectively, for compounds not in the learning sets. The good predictive ability of these models and their basis on simple to understand 2-D molecular fragments indicates their potential usefulness in computational screening methods for environmental estrogens. PMID- 15113070 TI - Predicting the biodegradation products of perfluorinated chemicals using CATABOL. AB - Perfluorinated chemicals (PFCs) form a special category of organofluorine compounds with particularly useful and unique properties. Their large use over the past decades increased the interest in the study of their environmental fate. Fluorocarbons may have direct or indirect environmental impact through the products of their decomposition in the environment. It is a common knowledge that biodegradation is restricted within non-perfluorinated part of molecules: however, a number of studies showed that defluorination can readily occur during biotransformation. To evaluate the fate of PFCs in the environment a set of principal transformations was developed and implemented in the simulator of microbial degradation using the catabolite software engine (CATABOL). The simulator was used to generate metabolic pathways for 171 perfluorinated substances on Canada's domestic substances list. It was found that although the extent of biodegradation of parent compounds could reach 60%, persistent metabolites could be formed in significant quantities. During the microbial degradation a trend was observed where PFCs are transformed to more bioaccumulative and more toxic products. Perfluorooctanoic acid and perfluorooctanesulfonate were predicted to be the persistent biodegradation products of 17 and 27% of the perfluorinated sulphonic acid and carboxylic acid containing compounds, respectively. PMID- 15113071 TI - The path to good aseptic guidance. PMID- 15113072 TI - Results of statistical analysis of blend and dosage unit content uniformity data obtained from the Product Quality Research Institute Blend Uniformity Working Group data-mining effort. PMID- 15113073 TI - Validation of microbial recovery from hydrogen peroxide-sterilized air. AB - The use of hydrogen peroxide as a sanitant in isolators and other barrier systems is well documented. To confirm that the isolator maintains a germ-free environment between decontamination cycles, microbiological air monitoring is performed after the sanitation and aeration cycles. In this study, we have shown that residual levels of hydrogen peroxide as low as 1 ppm can remain in the isolator and inhibit the growth of microorganisms after concentration on agar media. This lingering hydrogen peroxide can make accurate microbiological air monitoring difficult and can even cause false negative test results. To solve this issue, we have developed a new media that can mediate the effects of residual peroxide and prevent false negative test results. Initially, catalase was tested as a neutralizing agent but proved not to be efficient enough. Instead, 1% pyruvate was added, which was able to tolerate as much as 15 ppm Vaporous Hydrogen Peroxide (VHP) and ensured growth promotion of Staphylococcus aureus, Pseudomonas aeruginosa, Micrococcus luteus, Bacillus subtilis, Candida albicans and Aspergillus niger. The 1% pyruvate retained its neutralizing activity for Micrococcus luteus at up to 100 ppm VHP. Raising the pyruvate concentration to 5% pyruvate enabled neutralization of up to 300 ppm VHP, permitting subsequent growth of Micrococcus luteus on agar media. PMID- 15113074 TI - Factors affecting tailing in ethylene oxide sterilization part 1: When tailing is an artifact... and scientific deficiencies in ISO 11135 and EN 550. AB - Current approaches for validation of EtO sterilization cycles identified in ISO 11135 and EN 550 are scientifically deficient and result in full cycle times that are too short. This review article identifies the scientific flaws promulgated in the standards that lead to overestimation of process lethality. The reasons for these deficiencies are design-of-experiment errors that either fail to mathematically account for lag factors or that treat dissimilar samples as a single statistical sample set. Industry's movement toward parametric release, based on erroneous assumptions where exposure time rather than equivalent time is used, may create serious problems. The authors discuss these errors in detail, as well as reasonable methods for correcting them based on formulae for integrated lethality. Application of these methods will allow more accurate calculations of process lethality that are supported by biological data and appropriate mathematical analyses. PMID- 15113075 TI - Critical evaluation of the standard hydrolytic resistance test for glasses used for containers for blood and parenteral formulations. AB - As prescribed by pharmacopoeias, containers should meet certain condition of stability to be used for pharmaceutical products. Glass containers are classified according to their resistance to chemical attack, a test executed by heating the glass in contact with water for 30 min at 121 degrees C. The USP powdered glass test for glass containers was applied to different kinds of glasses used as containers for parenteral formulations. In this experiment not only the released alkalinity was measured but also the release of glass constituents: silicate, borate, sodium, and aluminum, and also the release of some impurities as copper and lead. The USP powdered glass test was also carried out with glass ampoules, clear and amber, in the presence of solution of some inorganic salts, NaCl, KCl, CaCl2, MgCl2, NaHCO3, NaH2PO4, KH2PO4, and sodium gluconate, citric acid and glucose. The results showed that even when releasing very low alkalinity, glasses also released their constituents, in concentration ranges from 8.8 to 33 mg/l for silicate, 0.9 to 6.9 mg/l for borate, 3 to 37 for mg/l for sodium and 0.5 to 2.4 mg/l for aluminum. More expressive results were found, however, for the tests done with solutions instead of pure water. The tests showed that, for most of the solutions, while the measured alkalinity was very low, high levels of the other constituents were found. Basic solutions of bicarbonate and gluconate presented the higher levels of all investigated constituents, confirming the ability of basic solutions to attack and dissolve the glass network. Glucose and citric acid interacted with the glass surface, selectively extracting aluminum, copper, and lead. Whereas silicate, borate and sodium found in these solutions were at levels similar to those found with pure water, the aluminum level was almost 20 times higher. This specific action of citrate and glucose could be related to their metal-complexing ability. The results indicate that even so-called "chemical resistant glasses," as measured by the hydrolytic resistance test, react with many substances when packaged in contact with them. The hydrolytic resistance test, when used as the sole measure of potential drug-container compatibility, is not reliable. PMID- 15113076 TI - Application of frequency-modulated spectroscopy in vacuum seal integrity testing of lyophilized biological products. AB - This study examined the feasibility of applying frequency-modulated spectroscopy (FMS) to test vacuum seal integrity of lyophilized protein pharmaceuticals in glass vials. A lyophilized recombinant monoclonal antibody was used as an example to demonstrate that FMS is a non-destructive method that could accurately and quickly determine vial vacuum integrity within a pressure range of 0.04 to 0.5 atm. The coefficient of determination (R2) of a bench-top instrument was found to be >0.99. Only seconds were required to analyze each sample. The instrument sensitivity and specificity were 0.95 and >0.99, respectively, based on analysis of approximately 40,000 samples. Because of low energy input by the instrument, no adverse effect on the protein quality was found immediately after up to 1 h of continuous laser exposure. The laser-exposed samples had comparable stability to non-exposed control vials after 12 weeks of storage at 40 degrees C. PMID- 15113077 TI - Nuclear organisation and subnuclear bodies. PMID- 15113078 TI - Searching for active ribosomal genes. PMID- 15113079 TI - Toxic RNA in the nucleus: unstable microsatellite expression in neuromuscular disease. PMID- 15113080 TI - Assembly and traffic of small nuclear RNPs. PMID- 15113081 TI - Intranuclear pre-mRNA trafficking in an insect model system. PMID- 15113082 TI - Photobleaching microscopy reveals the dynamics of mRNA-binding proteins inside live cell nuclei. PMID- 15113083 TI - Imaging of single mRNAs in the cytoplasm of living cells. PMID- 15113084 TI - Current concepts in the management of actinic keratosis. PMID- 15113085 TI - Peptide and amino acid glycation: new insights into the Maillard reaction. AB - Nonenzymatic glycation of proteins, peptides and other macromolecules (the Maillard reaction) has been implicated in a number of pathologies, most clearly in diabetes mellitus. but also in the normal processes of aging and neurodegenerative amyloid diseases such as Alzheimer's. In the early stage, glycation results in the formation of Amadori-modified proteins. In the later stages, advanced glycation end products (AGE) are irreversibly formed from Amadori products leading to the formation of reactive intermediates, crosslinking of proteins, and the formation of brown and fluorescent polymeric materials. Although, the glycation of structural proteins has been attributed a key role in the complications of diabetes, recent attention has been devoted to the physiological significance of glycated peptide hormones. This review focuses on the physico-chemical properties of the Amadori compounds of bioactive peptides of endogenous and exogenous origin, such as Leu-enkephalin and morphiceptin, investigated under different conditions as well as on novel pathways in the Maillard reaction observed from investigating intramolecular events in ester linked glycopeptides. PMID- 15113086 TI - Complete structure determination of the A chain of mistletoe lectin III from Viscum album L. ssp. album. AB - The complete primary structure of the A chain of mistletoe lectin III (ML3A), a type II ribosome-inactivating protein, was determined using proteolytic digests of ML3A, HPLC separation of the peptides, Edman degration and MALDI-MS. Based on our results, ML3A consists of 254 amino acid residues, showing a high homology to the A chain of isolectin ML1 with only 24 amino acid residue exchanges. A striking important structural difference compared with ML1A is the lack of the single N-glycosylation site in ML3A due to an amino acid exchange at position 112 (ML1A: NL112GS ==> ML3A: T112GS). The alignment of ML3A with the A chains of ML1, isoabrins, ricin D, Ricinus communis agglutinin and three lectins, identified from the Korean mistletoe Viscum album ssp. coloratum, demonstrates the rigid conservation of all amino acid residues, responsible for the RNA-N-glycosidase activity as reported for ricin D. In addition, the fully determined primary structure of ML3A will give further information about the biological mechanism of mistletoe lectin therapy. PMID- 15113087 TI - Metal ion-binding ability of tetrapeptides containing alpha-aminoisobutyric acid. AB - alpha-Aminoisobutyric acid (Aib), one of the Calpha,alpha-disubstituted glycines, is a sterically hindered amino acid that acts as a conformational constraint in peptides. However, studies for the application of the ability of Aib to control conformation are quite few. The paper focuses on the molecular recognition ability of acyclic oligopeptides containing Aib. Liquid-liquid extraction of nine kinds of metal ions from aqueous layers to nonpolar organic layers with acyclic tetrapeptides, X-Trp-Xaa2-Gly-Xaa4-NH-Ar (X = H or C6H5CH2OCO (Z), Xaa2 = Aib or Gly, Xaa4 = Leu or Ala, Ar = phenyl or 3,5-dimethylphenyl) was examined using picrate as the anion of ion pairs. The extraction behaviour of the metal ions with the tetrapeptides was investigated in the pH range from 3 to 9. In the case of basic pH regions, Cu(II) and Ag(I) were effectively extracted with Trp-Aib-Gly Leu-NH-Ar. Pd(II) was specifically extracted with Trp-Aib-Gly-Leu-NH-Ar in acidic pH regions. The extraction percent (%E) of the peptide host, which has a 3,5 dimethylphenyl group, was even larger than that of the host, which has a phenyl group. Moreover, Pd(II) was extracted with a peptide host which has Leu and a 3,5 dimethylphenyl group in the absence of picrate as the anion of ion pairs. The free alpha-amino group, the turn conformation and the hydrophobicity of peptide molecules were important factors for the extraction of the metals. PMID- 15113088 TI - Peptide helices with pendant cycloalkane rings. Characterization of conformations of 1-aminocyclooctane-1-carboxylic acid (Ac8c) residues in peptides. AB - A pentapeptide, Boc-Leu-Ac8c-Ala-Leu-Ac8c-OMe 1, an octapeptide, Boc-Leu-Ac8c-Ala Leu-Ac8c-Ala-Leu-Ac8c-OMe 2 and a tripeptide, Boc-Aib-Ac8c-Aib-OMe 3 containing the 1-aminocyclooctane-1-carboxylic acid residue (Ac8c) were synthesized and conformationally characterized by x-ray diffraction studies in the crystal state. Peptides 1 and 2 were also studied by NMR in CDC13 solution. Peptide 1 adopts a purely 3(10)-helical conformation in crystals, stabilized by three intramolecular 1 <-- 4 hydrogen bonds. Peptide 2 in crystals is largely 3(10)-helical with distortion in the backbone at the N-terminus by the insertion of a water molecule between Ac8c (2) CO and Ala (6) NH groups. Peptide 3 forms a C10-ring structure, i.e. a type III (III') beta- turn conformation stabilized by an intramolecular 1 <-- 4 hydrogen bond. Five cyclooctane rings assume boat-chair conformations, whereas the sixth [Ac8c(8) in 2] is appreciably distorted, resembling a chiral intermediate in the pseudorotational pathway from the boat-chair to the twisted boat-chair conformation. Internal bond angles of the cyclooctane rings are appreciably distorted from the tetrahedral value, a characteristic feature of the cyclooctane ring. Peptide 1 crystallized in the space group P212121 with a = 11.900(4) A, b = 18.728(6) A, c = 20.471(3) A and Z = 4. The final R1 and wR2 values are 0.0753 and 0.2107, respectively, for 3901 observed reflections [Fo > or = 3 sigma (Fo)]. Peptide 2 crystallized in space group P21 with a = 12.961(5) A, b = 17.710(10) A, c = 15.101(7) A, beta = 108.45(4) degrees and Z = 2. The final R1 and wR2 values are 0.0906 and 0.1832, respectively, for 2743 observed reflections [Fo > or = 3sigma (Fo)]. 1H-NMR studies on both the peptides strongly suggest the persistence of 3(10)-helical conformations in solution. Peptide 3 crystallized in the space group P21/n, with a = 10.018(1) A, b = 20.725(1) A, c = 12.915(1) A and Z = 4. The final R1 and wR2 values are 0.0411 and 0.1105, respectively, for 3634 observed reflections [Fo > or = 4sigma (Fo)]. PMID- 15113089 TI - Isolation of allicepin, a novel antifungal peptide from onion (Allium cepa) bulbs. AB - From the bulbs of the onion Allium cepa, a novel antifungal peptide distinct from the antimicrobial peptide previously reported from onion seeds was isolated. The antifungal peptide, designated allicepin, was purified with a procedure that involved aqueous extraction, ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel and FPLC-gel filtration on Superdex 75. Allicepin was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel. The molecular weight of allicepin was estimated to be 10 K by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration on Superdex 75. Allicepin exerted an inhibitory activity on mycelial growth in several fungal species including Botrytis cinerea, Fusarium oxysporum, Mycosphaerella arachidicola and Physalospora piricola. PMID- 15113090 TI - Growing plasma proteome sets stage for new diagnostics. PMID- 15113091 TI - Human liver proteome project. PMID- 15113092 TI - Proteomics and disease--the challenges for technology and discovery. PMID- 15113093 TI - Systems biology, proteomics, and the future of health care: toward predictive, preventative, and personalized medicine. AB - The emergence of systems biology is bringing forth a new set of challenges for advancing science and technology. Defining ways of studying biological systems on a global level, integrating large and disparate data types, and dealing with the infrastructural changes necessary to carry out systems biology, are just a few of the extraordinary tasks of this growing discipline. Despite these challenges, the impact of systems biology will be far-reaching, and significant progress has already been made. Moving forward, the issue of how to use systems biology to improve the health of individuals must be a priority. It is becoming increasingly apparent that the field of systems biology and one of its important disciplines, proteomics, will have a major role in creating a predictive, preventative, and personalized approach to medicine. In this review, we define systems biology, discuss the current capabilities of proteomics and highlight some of the necessary milestones for moving systems biology and proteomics into mainstream health care. PMID- 15113094 TI - Building a foundation for the human proteome: the role of the Human Proteome Organization. AB - Proteomics holds both substantial promise and substantial challenges. For proteomics to bear fruit on a large scale from a disease investigation point of view, it is essential to build a solid foundation for the field. Given the magnitude of the challenges, it is necessary to build a foundation by bringing together the private and public sectors. The Human Proteome organization is promoting the field of proteomics by engaging in such an effort and is developing several major initiatives. PMID- 15113095 TI - Proteomics strategies in cardiovascular research. AB - Cardiovascular research of the past decades dealt with classical pathophysiological descriptions, then shifted toward the identification of relevant receptors, and then proceeded to the analysis of signal transduction pathways. Most recently, hand in hand with the achievements of the human genome project, the research has gone down the road toward molecular biological "disease gene(s) mapping". The application of proteome research will attempt to close the gap between genomic (and genetic) analysis and the physiological research. The rich source of heart surgery specimens represents an excellent starting point in data acquisition of proteomic context. Furthermore, animal models of cardiovascular diseases and deficiencies are considered, and will be explored. Examples of results from feasibility studies are given, with the emphasis on quantitative evaluation of proteomic components, hoping to discover co-regulated sets of proteins that are involved in any particular disease state. Identification of new, not yet discovered proteins will be pursued, though the emphasis of this work will be on the definition of characteristic sets of expressed proteins, which in turn might be able to delimit the state of disease and prognosis of therapy outcome. Besides the systematic issues, this paper refers to a number of methodological questions, like the comparison of the proteins detected by staining procedures and proteins detected in models in which biosynthetic labeling is applicable. PMID- 15113096 TI - Clinical proteomics: revolutionizing disease detection and patient tailoring therapy. AB - The evolving discipline of Clinical Proteomics is more than simply describing and enumerating the systematic changes in the protein constituency of a cell, or just generating lists of proteins that increase or decrease in expression as a cause or consequence of disease. Clinical applications of proteomics involve the use of proteomic technologies at the bedside with the ultimate goal to characterize the information flow through the intra- and extracellular molecular protein networks that interconnect organ and circulatory systems together. These networks are both new targets for therapeutics themselves as well as underpin the dynamic changes that give rise to cascades of new diagnostic biomarkers. The analysis of human cancer can be used as a model for how clinical proteomics is having an impact at the bedside for early detection, rational therapeutic targeting, and patient tailored therapy. PMID- 15113097 TI - Mining disease susceptibility genes through SNP analyses and expression profiling using MALDI-TOF mass spectrometry. AB - To find genes that underlie disease susceptibilities, genome-wide single nucleotide polymorphisms (SNPs) have been analyzed using high-throughput matrix assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS). As a proof-of-concept for this approach, gene regions have been identified that were previously associated by others with certain diseases or traits. On the same technology platform, accurate and absolute transcriptional profiling can be performed and applied to allele expression analysis. Here, we provide a brief review of the technology and its applications to disease gene discovery. PMID- 15113098 TI - An effective and rapid method for functional characterization of immunoadsorbents using POROS beads and flow cytometry. AB - To facilitate the construction, functional characterization, and use of immunoadsorbents, we have developed a flow cytometry method that allows rapid assessment of large numbers of particle-bound antibodies. Protein G derivitized POROS beads were used to bind affinity-purified antibodies specific for synthetic peptides designed from human plasma proteins. The antibodies were covalently coupled to the beads and used to capture and release synthetic peptides that had been labeled at the C-terminus with the fluorochrome Alexa Fluor 488. Antibody coupling and specificity of antigen binding and release were measured by analysis of the POROS affinity beads by flow cytometry. The affinity-capture matrixes were also used through several antigen-binding and release cycles without loss of peptide binding efficiency. The ability to produce and characterize extremely small amounts of POROS affinity matrices will facilitate their use in protein microchemical procedures such as protein chip technology, monoclonal antibody screening and mass spectrometry, applications where analytes are limiting or present in low abundance in complex mixtures. PMID- 15113099 TI - Mass spectrometric quantitation of peptides and proteins using Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA). AB - A method (denoted SISCAPA) for quantitation of peptides in complex digests is described. In the method, anti-peptide antibodies immobilized on 100 nanoliter nanoaffinity columns are used to enrich specific peptides along with spiked stable-isotope-labeled internal standards of the same sequence. Upon elution from the anti-peptide antibody supports, electrospray mass spectrometry is used to quantitate the peptides (natural and labeled). In a series of pilot experiments, tryptic test peptides were chosen for four proteins of human plasma (hemopexin, alpha1 antichymotrypsin, interleukin-6, and tumor necrosis factor-alpha) from a pool of 10,203 in silico tryptic peptide candidates representing 237 known plasma components. Rabbit polyclonal antibodies raised against the chosen peptide sequences were affinity purified and covalently immobilized on POROS supports. Binding and elution from these supports was shown to provide an average 120-fold enrichment of the antigen peptide relative to others, as measured by selected ion monitoring (SIM) or selected reaction monitoring (SRM) electrospray mass spectrometry. The columns could be recycled with little loss in binding capacity, and generated peptide ion current measurements with cycle-to-cycle coefficients of variation near 5%. Anti-peptide antibody enrichment will contribute to increased sensitivity of MS-based assays, particularly for lower abundance proteins in plasma, and may ultimately allow substitution of a rapid bind/elute process for the time-consuming reverse phase separation now used as a prelude to online MS peptide assays. The method appears suitable for rapid generation of assays for defined proteins, and should find application in the validation of diagnostic protein panels in large sample sets. PMID- 15113100 TI - Assessing protein patterns in disease using imaging mass spectrometry. AB - Direct tissue profiling and imaging mass spectrometry (MS) provides a detailed assessment of the complex protein pattern within a tissue sample. MALDI MS analysis of thin tissue sections results in over of 500 individual protein signals in the mass range of 2 to 70 kDa that directly correlate with protein composition within a specific region of the tissue sample. To date, profiling and imaging MS has been applied to multiple diseased tissues, including human gliomas and nonsmall cell lung cancer. Interrogation of the resulting complex MS data sets has resulted in identification of both disease-state and patient-prognosis specific protein patterns. These results suggest the future usefulness of proteomic information in assessing disease progression, prognosis, and drug efficacy. PMID- 15113101 TI - Implications of new proteomics strategies for biology and medicine. AB - Advances in proteomics have fundamentally changed the paradigm of discovery for drug targets and novel biomarkers. Proteomics methodologies currently used will be reviewed in this paper, including structural proteomics, quantitative proteomics, and functional proteomics. A strategy to identify differentially expressed cell surface proteins as monoclonal therapeutic targets in oncology will be discussed. PMID- 15113102 TI - Development of natural protein microarrays for diagnosing cancer based on an antibody response to tumor antigens. AB - The detection of autoantibodies to tumor antigens has potential utility for the early diagnosis of cancers. In previous studies, we have identified tumor antigens based on Western blot analysis of tumor cell lysates that were incubated with subject sera to identify proteins that elicit specific reactivity in sera from patients with the corresponding tumor type. More recently, we have explored the use of microarrays spotted with tumor proteins as an alternative to Western blots. Microarrays provide a high throughput, high sensitivity alternative to the use of Western blots for tumor antigen profiling. In this study, we have assessed the reproducibility of natural protein microarrays and their ability to distinguish between lung cancer sera and controls. Protein lysates from the A549 human lung adenocarcinoma cell line were separated into 1840 fractions that were spotted in duplicate, along with various controls, on nitrocellulose coated slides. Sera from 18 newly diagnosed patients with lung cancer and from 15 healthy controls were each hybridized to an individual microarray. The reactivity of arrayed proteins with Ig was determined by incubation with biotinylated goat anti-human-Ig followed by phycoerythrin-conjugated streptavidin. The intensity measures of duplicate spots (within-slide) and duplicate slides (between-slides) were highly reproducible, exhibiting correlation values >0.9. A total of 63 of the 1840 arrayed fractions demonstrated increased reactivity in cancer patients relative to controls as measured by a rank-based statistic (p < 0.008). Microarrays of tumor-derived proteins provide the means for uncovering a repertoire of tumor antigens that have induced an antibody response in patients with specific cancers. PMID- 15113103 TI - Chemistry-based functional proteomics: mechanism-based activity-profiling tools for ubiquitin and ubiquitin-like specific proteases. AB - Determining the biological function of newly discovered gene products requires the development of novel functional approaches. To facilitate this task, recent developments in proteomics include small molecular probes that target proteolytic enzyme families including serine, threonine, and cysteine proteases. For the families of ubiquitin (Ub) and ubiquitin-like (UBL)-specific proteases, such tools were lacking until recently. Here, we review the advances made in the development of protein-based active site-directed probes that target proteases specific for ubiquitin and ubiquitin-like proteins. Such probes were applied successfully to discover and characterize novel Ub/UBL-specific proteases. Ub/UBL processing and deconjugation are performed by a diverse set of proteases belonging to several different enzyme families, including members of the ovarian tumor domain (OTU) protease family. A further definition of this family of enzymes will benefit from a directed chemical proteomics approach. Some of the Ub/UBL-specific proteases react with multiple Ub/UBLs and members of the same protease family can recognize multiple Ub/UBLs, underscoring the need for tools that appropriately address enzyme specificity. PMID- 15113104 TI - Proteomics: is it an approach to understand the progression of chronic lung disorders? AB - Tissue injury, mediated by pathologically elevated production and action of various serine- and matrix metalloproteinases (MMPs), is a hallmark of chronic inflammatory airway diseases (CIAD). CIAD includes such diseases as bronchial asthma, bronchiectasis, and chronic obstructive pulmonary disease. Tissue injury, as a consequence of chronic inflammation, can disturb the relevant repair mechanisms and also result in irreversible alteration of lung architecture. By use of proteomic methods, we analyzed proteinase cascades as an initiator of tissue destruction in CIAD. The present results revealed that elevated levels of MMP-8, -13, -14, and -2, mainly in active forms, can also be detected in CIAD BALFs. Enhanced levels of different active MMPs evidently reflect ongoing tissue destructive inflammation and airway remodeling occurring in CIAD lung. An inverse correlation between BALF MMP-8 levels and activation degree and airflow obstruction in bronchial asthma tissue injury was shown for the first time. This strongly indicates that chronic peri-inflammatory tissue injury is a main cause of decline of lung functional capacity. Together, these data suggest that the serine and MMP proteinase network is an important feature in predicting clinical worsening of airway obstruction in CIAD. Activation of elevated MMPs seems to have a common profile for all studied CIAD, but different lung disorders react differently to ICS treatment. PMID- 15113105 TI - Use of proteomics to discover novel markers of cardiac allograft rejection. AB - Endomyocardial biopsy remains the most reliable method of detecting rejection following cardiac transplantation. Despite numerous attempts to detect rejection using a blood assay, none have proved reliable enough to replace the biopsy. Here, we have investigated the hypothesis that proteomics has the potential to reveal many molecules which are upregulated in the heart during rejection, some of which may serve as novel blood markers of rejection. Initially, sequential cardiac biopsies (33 in total) from 4 patients were analysed by two-dimensional gel electrophoresis according to whether they showed rejection (n = 16) or no rejection (n = 17); over 100 proteins were found to be upregulated by between 2- and 50-fold during rejection. Of these, 13 were identified and were found to be cardiac specific or heat shock proteins. Two of these (alphaB-crystallin, tropomyosin) were measured by ELISA in the sera of 17 patients followed for 3 months after their transplants. Mean levels of alphaB-crystallin and tropomyosin were significantly higher in sera associated with biopsies showing 1A (p = 0.007) or all grades of rejection (p = 0.022) compared to no rejection. These studies demonstrate that proteomics is a powerful method that can be used to identify novel serum markers of human cardiac allograft rejection. PMID- 15113106 TI - Molecular profiling of experimental Parkinson's disease: direct analysis of peptides and proteins on brain tissue sections by MALDI mass spectrometry. AB - Direct molecular profiling of biological samples using matrix-assisted laser desorption ionization mass spectrometry is a powerful tool for identifying phenotypic markers. In this report, protein profiling was used for the first time to generate peptide and protein profiles of brain tissue sections obtained from experimental Parkinson's disease (unilaterally 6-hydroxydopamine treated rats). The mass spectrometer was used to map the peptide and protein expression directly on 12 microm tissue sections in mass-to-charge (m/z) values, providing the capability of mapping specific molecules of the original sample, that is, localization, intensity and m/z ratio. Several protein expression profile differences were found in the dopamine depleted side of the brain when compared to the corresponding intact side, for example, calmodulin, cytochrome c, and cytochrome c oxidase. An increased ratio of post-translational modifications such as acetylations were found in the striatum of proteins in the dopamine depleted side of the brain. These modifications were decreased after subchronic administration of L-Dopa. The present study shows that unique protein profiles can be obtained in specific brain regions (and subregions) directly on brain tissue sections and allows for the study of complex biochemical processes such as those occurring in experimental Parkinson's disease. PMID- 15113107 TI - Proteomics in malaria. AB - The recent completion of human, Anopheles gambiae, and Plasmodium falciparum genomes relevant to the study of human malaria allows the application of modern proteomic technologies to complement previously implemented conventional approaches. Proteomic analysis has been employed to elucidate global protein expression profiles, subcellular localization of gene products, and host-pathogen interactions that are central to disease pathogenesis and treatment. The high throughput nature of these techniques is in accord with the pace of drug and vaccine development that have the potential to directly reduce the morbidity and mortality of disease. PMID- 15113108 TI - Exploring the context of the lung proteome within the airway mucosa following allergen challenge. AB - The lung proteome is a dynamic collection of specialized proteins related to pulmonary function. Many cells of different derivations, activation states, and levels of maturity contribute to the changing environment, which produces the lung proteome. Inflammatory cells reacting to environmental challenge, for example from allergens, produce and secrete proteins which have profound effects on both resident and nonresident cells located in airways, alveoli, and the vascular tree which provides blood cells to the parenchyma alveolar bed for gas exchange. In an experimental model of allergic airway inflammation, we have compared control and allergen challenged lung compartments to determine global protein expression patterns using 2D-gel electrophoresis and subsequent spot identification by MS/MS mass spectrometry. We have then specifically isolated the epithelial mucosal layer, which lines conducting airways, from control and allergen challenged lungs, using laser capture technology and performed proteome identification on these selected cell samples. A central component of our investigations has been to contextually relate the histological features of the dynamic pulmonary environment to the changes in protein expression observed following challenge. Our results provide new information of the complexity of the submucosa/epithelium interface and the mechanisms behind the transformation of airway epithelium from normal steady states to functionally activated states. PMID- 15113109 TI - Development drivers. PMID- 15113110 TI - Speciation of heavy metals in landfill leachate: a review. AB - The literature was reviewed with respect to metal speciation methods in aquatic samples specifically emphasizing speciation of heavy metals in landfill leachate. Speciation here refers to physical fractionation (particulate, colloidal, dissolved), chemical fractionation (organic complexes, inorganic complexes, free metal ions), as well as computer-based thermodynamic models. Relatively few landfill leachate samples have been speciated in detail (less than 30) representing only a few landfills (less than 15). This suggests that our knowledge about metal species in landfill leachate still is indicative. In spite of the limited database and the different definitions of the dissolved fraction (< 0.45 microm or < 0.001 microm) the studies consistently show that colloids as well as organic and inorganic complexes are important for all heavy metals in landfill leachate. The free metal ion constitutes less than 30%, typically less than 10%, of the total metal concentration. This has significant implications for sampling, since no standardized procedures exist, and for assessing the content of metals in leachate in the context of its treatment, toxicity and migration in aquifers. PMID- 15113111 TI - Speciation of lipids and humus-like colloidal compounds in a forest soil reclaimed with municipal solid waste compost. AB - The progressive transformations of lipid and humus-like fractions in soil after massive input (400 Mg ha(-1)) of urban waste have been studied during an 87-week experiment in field plots of a degraded Calcic Regosol in Central Spain. Structural changes in the macromolecular fractions were small when compared with the qualitative and quantitative changes in lipid composition. The intense depletion of the lipid fraction with time and the decrease of the humic acid to fulvic acid ratio were the most significant quantitative indices of the compost transformation in soil. Changes in soil lipid fractions were especially noted in relation to their speciation status and distribution patterns (carbon preference index and relative chain length). Three subfractions were considered: (I) direct extraction with petroleum ether, (II) liquid-liquid extraction after soil treatment with 2 M H3PO4 and (III) after soil treatment with 0.1 M NaOH. Although lipid concentration tends to decrease with time, lipids in the fraction tightly bonded to soil (III) remained qualitatively and quantitatively constant in the course of the field experiment. Gas chromatographic-mass spectrometric analyses showed that the more stable the association of lipid to the soil matrix, the fewer the changes observed in the distribution pattern of the fatty acids during the progressive transformation stages. PMID- 15113112 TI - Efficiency of the anaerobic treatment of the organic fraction of municipal solid waste: collection and pretreatment. AB - This report is based on several years of co-operation between our research groups and Danish biogas plants. Throughout the years, there has been a fruitful exchange of know-how and experiences in laboratory scale on the one hand and large scale on the other, leading to a better understanding of the principles of the anaerobic digestion process and to an optimization of its large-scale implementation. In order to get an overview of the current situation concerning the treatment of the organic fraction of municipal solid waste (OFMSW) in Denmark, interviews were carried out with operators of the biogas plants where OFMSW is treated and the municipality staff responsible for waste management. With the aim of fulfilling the governmental goal to treat 150,000 tons of OFMSW by the year 2004 mainly by anaerobic digestion, the different municipalities are investigating different concepts of waste collection and treatment. The quality of the OFMSW treated is the key to smooth operation of the biogas process including a high biogas yield and production of an effluent that is feasible for use as fertilizer on agricultural land. Comparison of the different concepts leads to the conclusion that source-sorting of OFMSW in paper bags is preferable to collection in plastic bags and successive separation of plastics in a waste processing treatment plant. PMID- 15113113 TI - Assessment of the methane oxidation capacity of soil. AB - Methane oxidation capacity of three soil matrices was assessed using a new method based on the pressure fall resulting from methane oxidation. The effects of incubation temperature, moisture and methane content on the oxidation capacity were studied. Parameters observed were non-methanotrophic oxygen consumption, methane assimilation, length of the lag period before steady pressure fall was reached, initial pressure fall and maximum pressure fall. Filling material taken from a biofilter was tested. The length of the lag period was an index of prior exposure of the soil to methane emissions. Incubation temperature of 30 degrees C and soil moisture at the water-holding capacity were chosen to standardize the test. Oxygen depletion by non-methanotrophs varied between 40 and 60% of the total oxygen consumption while methane assimilation was at 40 to 50% of the total methane consumption. PMID- 15113114 TI - Thermal treatment of stabilized air pollution control residues in a waste incinerator pilot plant. Part 1: Fate of elements and dioxins. AB - Air pollution control (APC) residues from municipal solid waste incinerator plants that are treated by means of the Ferrox process can be more safely disposed of due to reduction of soluble salts and stabilization of heavy metals in an iron oxide matrix. Further stabilization can be obtained by thermal treatment inside a combustion chamber of a municipal solid waste incinerator. The influence of the Ferrox products on the combustion process, the quality of the residues, and the partitioning of heavy metals between the various solids and the gas have been investigated in the Karlsruhe TAM-ARA pilot plant for waste incineration. During the experiments only few parameters were influenced. An increase in the SO2 concentration in the raw gas and slightly lower temperatures in the fuel bed could be observed compared with reference tests. Higher contents of Fe and volatile heavy metals such as Zn, Cd, Pb and partly Hg in the Ferrox products lead to increased concentration of these elements in the solid residues of the co-feeding tests. Neither the burnout nor the PCDD/F formation was altered by the addition of the Ferrox products. Co-feeding of treated APC residues seems to be a feasible approach for obtaining a single solid residue from waste incineration. PMID- 15113115 TI - Thermal treatment of stabilized air pollution control residues in a waste incinerator pilot plant. Part 2: Leaching characteristics of bottom ashes. AB - With the perspective of generating only one solid residue from waste incineration, co-feeding of municipal solid waste and air pollution control residues stabilized by the Ferrox process was investigated in the TAMARA pilot plant incinerator as described in Bergfeldt et al. (Waste Management Research, 22, 49-57, 2004). This paper reports on leaching from the combined bottom ashes. Batch leaching test, pH-static leaching tests, availability tests and column leaching tests were used to characterize the leaching properties. The leaching properties are key information in the context of reuse in construction or in landfilling of the combined residue. In general, the combined bottom ashes had leaching characteristics similar to the reference bottom ash, which contained no APC residue. However, As and Pb showed slightly elevated leaching from the combined bottom ashes, while Cr showed less leaching. The investigated combined bottom ashes had contents of metals comparable to what is expected at steady state after continuous co-feeding of APC residues. Only Cd and Pb were partly volatilized (30-40%) during the incineration process and thus the combined bottom ashes had lower contents of Cd and Pb than expected at steady state. Furthermore, a major loss of Hg was, not surprisingly, seen and co-feeding of Ferrox-products together with municipal solid waste will require dedicated removal of Hg in the flue gas to prevent a build up of Hg in the system. In spite of this, a combined single solid residue from waste incineration seems to be a significant environmental improvement to current technology. PMID- 15113116 TI - Structure and composition of tubular aggregates of skeletal muscle fibres. AB - Unusual regions of densely packed membranous tubules known as tubular aggregates (TAs) have been observed in skeletal muscle fibres of mammals under numerous pathological conditions but also in health. Their causality is unclear. It is neither known whether TAs are destructive and should be treated or whether they have a compensating function in an endangered muscle. In spite of many similarities, the histochemical, immunocytochemical and ultrastructural characteristics of tubular aggregates do vary. Histochemistry provided an overall characteristic of TAs as membranous inclusions with a variety of enzymatic activities. Immunocytochemical evidence revealed that tubular aggregates contain miscellaneous proteins and that derive from membranes of sarcoplasmic reticulum and mitochondria. No evidence for the presence of contractile and cytoskeletal proteins in TAs was found. Ultrastructurally, TAs are characterized as more or less densely packed aggregates of vesicular or tubular membranes of variable forms and sizes that may contain amorphous material, filaments or inner tubules. Various reported types of tubular aggregates, namely, proliferating terminal cisterns, vesicular membrane collections, TAs with double-walled tubules, TAs with single-walled tubules, aggregates of dilated tubules with inner tubules, aggregates of tubulo-filamentous structures, filamentous tubules, riesentubuli, and related membranous structures including cylindrical spirals are sumarized and analyzed here in detail. PMID- 15113117 TI - Inactivation of L-type calcium channels in cardiomyocytes. Experimental and theoretical approaches. AB - The L-type calcium current (ICa) plays an important role in excitation contraction coupling of heart cells. It is critical for forming the major trigger for Ca(2+)-induced Ca(2+) release from the sarcoplasmic reticulum and hence its feedback regulation is of fundamental biological significance. The channel inactivation sharpens the kinetics and temporal precision of the Ca(2+) signals so that it prevents longer-term increases in free intracellular Ca(2+) concentration. Cardiac L-type Ca(2+) channels are known to inactivate through voltage- and Ca(2+)-dependent mechanisms. Pure voltage-dependent inactivation has a much slower time course of development than Ca(2+)-dependent inactivation and plays minor role in inhibition of Ca(2+) influx into the cell. The major determinant of the inactivation kinetics of Ca(2+) current during depolarization is Ca(2+)-dependent mechanisms. Furthermore, it is possible to distinguish two phases in Ca(2+)-dependent inactivation of calcium current: a slow phase that depends on Ca(2+) flow through the channels (Ca(2+) current-dependent inactivation) and a fast one that depends on Ca(2+) released from the sarcoplasmic reticulum (Ca(2+) release-dependent inactivation). Although both Ca(2+) released from the SR and Ca(2+) permeating channels play a role, SR released Ca(2+) is the most effective inactivation mechanism in inhibition of Ca(2+) entry through the channel. PMID- 15113118 TI - Evaluation of human erythrocytes as model cells in photodynamic therapy. AB - The role of erythrocytes as targets in photodynamic therapy is a controversially discussed topic in the literature. Therefore five different, but well known photosensitisers (three zinc phthalocyanines, tetrabenzoporphine and pheophorbide a delivered in liposomes were used for photodynamic treatment of human erythrocytes. The phototoxic effect on these cells showed pronounced differences. It was in the range: zinc phthalocyanine = pheophorbide a > tetrabenzoporphine >> zinc octa-n-alkyl phthalocyanines. Data from the zinc octa-n-alkyl phthalocyanines were compared with photodynamic effects within cutaneous cell lines, treated under the same experimental conditions. The results show that erythrocytes are unlikely to make good models for predicting the efficiency of the photosensitiser in general, and the same applies to cells other than erythrocytes and in vivo. Possible reasons could be differences in dye accumulation. However, erythrocytes may well serve as model cells to explore the cellular and molecular mechanisms of photodynamic treatment. PMID- 15113119 TI - Effects of oxidation on changes of compressibility of bovine serum albumin. AB - The methods of ultrasound velocity and density measurements were used to study the adiabatic compressibility of bovine serum albumin (BSA) during its oxidation by the prooxidants Cu2+ and 2,2'-azobis(2-amidinopropane) hydrochloride (AAPH). We did not find changes of compressibility of BSA in the presence of copper ions at rather high molar ratio Cu2+/BSA = 0.66 mol/mol. This can be explained by binding of the Cu2+ to the binding site of BSA and thus protecting the prooxidant action of the copper. However, AAPH-mediated oxidation of BSA resulted in an increase of its apparent specific compressibility (psik/beta0). These changes could be caused by the fragmentation of the protein. PMID- 15113120 TI - Developmental changes in uncoupling protein 1 and F1-ATPase subunit levels in the golden hamster brown adipose tissue mitochondria as determined by electron microscopy in situ immunocytochemistry. AB - The postnatal developmental changes in mitochondrial uncoupling protein 1 (UCP 1) and F1-ATP synthase (ATPase) subunit levels in the interscapular brown adipose tissue (BAT) were studied in golden Syrian hamsters (Mesocricetus auratus) using electron microscopy in situ immunocytochemistry. The relatively low initial density of 5 nm gold conjugated anti-UCP 1 immunocomplexes gradually increased from 7- to 21-day-old animals and numerous immunocomplexes were found on the mitochondrial membranes of adult hamsters. At the age of 7-9 days, a positive reaction was also detected in the cytoplasm of BAT adipocytes. Immunolocalization of F1-ATPase subunit indicated its presence in BAT mitochondria and cytoplasm of 7- to 9-day-old animals. However, contrary to UCP 1, intensity of the immunostaining of F1-ATPase subunit rapidly decreased both in mitochondria and cytoplasm between the 10th and 21st postnatal day and it became stabilized in adult animals at a very low level restricted to mitochondria. These results confirm that profound changes in the enzymatic apparatus of BAT mitochondrial membranes, leading to formation of thermogenic mitochondria, occur not until the early postnatal period of hamster ontogenetic development. PMID- 15113121 TI - Mechanisms involved in the increase in intracellular calcium following hypotonic shock in bovine articular chondrocytes. AB - The extracellular osmotic environment of chondrocytes fluctuates during joint loading as fluid is expressed from and reimbibed by the extracellular matrix. Matrix synthesis by chondrocytes is modulated by joint loading, possibly mediated by variations in intracellular composition. The present study has employed the Ca2+-sensitive fluoroprobe Fura-2 to determine the effects of hypotonic shock (HTS) on intracellular Ca2+ concentration ([Ca2+]i) and to characterise the mechanisms involved in the response for isolated bovine articular chondrocytes. In cells subjected to a 50% dilution, [Ca2+]i rapidly increased by approximately 250%, a sustained plateau being achieved within 300 s. The effect was inhibited by thapsigargin or by removal of extracellular Ca2+, indicating that the rise in [Ca2+]i reflects both influx from the extracellular medium and release from intracellular stores. Inhibition of the response by neomycin implicates activation of PLC and IP3 synthesis in the mobilisation of Ca2+ from intracellular stores. The rise was insensitive to inhibitors of L-type voltage activated Ca2+ channels (LVACC) or reverse mode Na+/Ca2+ exchange (NCE) but could be significantly attenuated by ruthenium red, an inhibitor of transient receptor potential vanilloid (TRPV) channels and by Gd3+, a blocker of stretch-activated cation (SAC) channels. The HTS-induced rise in [Ca2+]i was almost completely absent in cells treated with Ni2+, a non-specific inhibitor of Ca2+ entry pathways. We conclude that in response to HTS the opening of SACC and a member of TRPV channel family leads to Ca2+ influx, simultaneously with the release from intracellular stores. PMID- 15113122 TI - Positive inotropic effect of the inhibition of cyclic GMP-stimulated 3',5'-cyclic nucleotide phosphodiesterase (PDE2) on guinea pig left atria in eu- and hyperthyroidism. AB - The significance of PDE2 on the atrial inotropy was studied in eu- and hyperthyroidism. The contractile force was measured and negative inotropic capacity of N6-cyclopentyladenosine (CPA) was determined on left atria isolated from 8-day thyroxine- or solvent-treated guinea pigs, in the presence or absence of EHNA (adenosine deaminase and PDE2 inhibitor) or NBTI (nucleoside transporter inhibitor). EHNA was administered to inhibit PDE2, while NBTI was used to model the accumulation of endogenous adenosine. The reduction of the contractile force caused by EHNA was smaller in the thyroxine-treated atria than in the solvent treated samples. Contrary, NBTI induced a decrease in the contractile force without significant difference between the two groups. In addition, EHNA enhanced the efficiency of CPA in thyroxine-treated atria and did not affect it in solvent treated samples, while the response to CPA was decreased by NBTI in all atria, especially in hyperthyroidism. On the basis of greater retention of the contractile force and sustained/enhanced responsiveness to CPA in the presence of EHNA we conclude that PDE2's inhibition has a significant positive inotropic effect in guinea pig atria and this effect is proven to be augmented in hyperthyroidism. PMID- 15113123 TI - Inhibition of Cav3.1 channel by silver ions. AB - We have investigated the effects of AgCl and AgNO3 on the Cav3.1 calcium channels stably expressed in the HEK 293 cells. Ca2+ was used as a charge carrier. Both forms of Ag+ blocked the Cav3.1 channel and negatively shifted the I-V relations in a concentration-dependent manner. The inhibition of current amplitude by AgCl was voltage-dependent and increased with increasing amplitude of the depolarizing pulse. Furthermore, AgCl but not AgNO3 accelerated the kinetics of current activation. No effect on current inactivation or steady-state inactivation of the channel was observed for AgCl or AgNO3. PMID- 15113124 TI - Intimate contacts of mitochondria with nuclear envelope as a potential energy gateway for nucleo-cytoplasmic mRNA transport. AB - The aim of the present study was to show that close contacts of mitochondria with nuclear envelope need not be just an accidental situation in the cell, but that such contacts could serve for flow of energy from a place of origin directly to a place of consumption. Mitochondria in close proximity to the nuclear envelope can be found virtually in all metabolically active cells. We used transmission electron microscopy to demonstrate this entity in different leukemia cells of human origin (patient's blood) and in mouse leukemia cell line. At high resolution, not only close proximity but even fusion of mitochondrial and nuclear membranes can be seen. Based on available data about mRNA transport through the nuclear pore complex and observed contacts of mitochondria with nuclei, we hypothesize that such contacts can provide a gateway for energy delivery to power mRNA export from the nucleus to the cytoplasm. Hence the lumen of the nuclear envelope can serve for transport or even storage of macroergic molecules in a manner similar to sarcoplasmic reticulum in fast-twitch skeletal muscles. PMID- 15113125 TI - Effects of simvastatin treatment on oxidant/antioxidant state and ultrastructure of diabetic rat myocardium. AB - In the present study we investigated the effects of simvastatin treatment on lipid metabolism and peroxidation, antioxidant enzyme activities and ultrastructure of the diabetic rat myocardium. Diabetes was induced by single injection of streptozotocin (45 mg/kg i.p.). Eight weeks after induction of diabetes, a subgroup of control and of diabetic rats was treated with simvastatin for 4 weeks (10 mg/kg/day, orally). Blood glucose, plasma cholesterol and triacylglycerol, as well as levels of cardiac thiobarbituric acid reactive substances (TBARS) were significantly increased in diabetic rats. The activities of antioxidant enzymes, catalase (CAT) and glutathione peroxidase (GSHPx), were also elevated in the diabetic myocardium. Treatment with simvastatin markedly reduced serum triacylglycerol and cholesterol, and partially controlled hyperglycemia in diabetic animals. The increased activation of antioxidant enzymes and the excess of lipid peroxidation measured by TBARS were completely reversed by simvastatin treatment. Diabetic rats displayed ultrastructural ischemia-like alterations of cardiomyocytes and capillaries, which support oxidative stress-induced tissue remodelling. In the diabetic myocardium simvastatin treatment partly attenuated angiopathic and atherogenic processes, detected by electron microscopy. These results suggest that simvastatin, known as a lipid-lowering drug, may positively affect diabetes induced cardiovascular complications via reducing risks of atherosclerotic pathological processes, such as imbalance between oxidant and antioxidant state. PMID- 15113126 TI - High frequency of GJB2 mutation W24X among Slovak Romany (Gypsy) patients with non-syndromic hearing loss (NSHL). AB - Mutations in the GJB2 gene (connexin 26) represent a major cause of autosomal recessive non-syndromic hearing loss (NSHL) worldwide. In most Caucasian populations, the 35delG mutation in this gene was found to account for up to 50% of cases of the genetic non-syndromic childhood deafness. In populations of non European ethnic background, other GJB2 gene mutations are occasionally common, e.g. 167delT in Ashkenazi Jews, R143W in Africaans and 235delC in Koreans. In this work, DNA samples from 54 unrelated NSHL patients from endogamous and inbred population of Slovak Roms (Gypsies) from Eastern Slovakia were screened for GJB2 mutations. The coding region of the GJB2 gene of patients was sequenced and mutations W24X, R127H, V153I, L90P and V37I were found. In Slovak Romany population, mutation W24X accounts for 23.2%, R127H for 19.4%, 35delG for 8.3%, V153I for 3.7%, L90P for 3.7% and V37I for 0.9% of screened chromosomes. As the W24X mutation was previously found in India and Pakistan, were from the European Romanies originate, it was brought by the European Romnanies from their Indian homeland. The carrier frequency of 35delG was estimated for Slovak non-Romany population to be 3.3%, and for Slovak Romany population to 0.88%. The carrier frequency of W24X varied in different Slovak Romany subpopulations from 0.0% up to 26.1%. PMID- 15113127 TI - Low number of insulin receptors but high receptor protein content in adipose tissue of rats with monosodium glutamate-induced obesity. AB - In order to better understand the mechanisms leading to insulin resistance, the number of fat tissue insulin receptors, their affinity and insulin receptor protein in rats with monosodium glutamate-induced obesity were studied. Obese rats displayed significantly lower number of insulin receptors with high affinity. Surprisingly, the amount of insulin receptor protein was significantly elevated in these animals. The same relations have been already reported for angiotensin II binding and AT1 receptor protein in the same model of obesity. Therefore we suggest an existence of general defect of adipocyte cell membrane in monosodium glutamate-induced obesity characterized by the presence of high quantity of impaired receptor protein. PMID- 15113128 TI - Reduction of 1-methyl-1-nitrosourea-induced tumor burden with DNA vaccines encoding mutated ras epitopes and the costimulatory molecule B7.1. AB - 1-methyl-1-nitrosourea (MNU), a well characterized carcinogen, was used to induce adenocarcinomas in rat mammary gland. 150 days after the first injection of MNU, the animals were treated with DNA minigene vaccines encoding ras T cell epitopes together with the co-stimulatory molecule B7.1 (CD 80). Five injections with a biolistic device (gene gun) in monthly intervals significantly reduced the tumor burden. A therapeutic effect could be measured with both, DNA vaccines encoding ras epitopes and B7.1, as well as with a DNA vaccine expressing solely the B7.1 molecule thus indicating the potential of genetic vaccination for turnor treatment. PMID- 15113129 TI - Proteins released from liver after ischaemia induced an elevation of heart resistance against ischaemia-reperfusion injury: 1. Beneficial effect of protein fraction isolated from perfusate after ischaemia and reperfusion of liver. AB - OBJECTIVES: Numerous mechanisms have been proposed to participate in adaptation of heart to ischaemia by ischaemic preconditioning. We have described previously a release of cardio-protective protein fraction during ischaemic preconditioning of dog heart. In the current study the effect of high soluble protein fraction (HS fraction) released from isolated perfused rat liver after ischaemia and reperfusion was examined on isolated perfused rat heart during ischaemia reperfusion injury. METHODS: Livers were subjected to 30 or 60 min ischaemia followed with 120 min reperfusion. HS fraction was isolated using ammonium sulphate precipitation and dissolved in perfusion solution before Langendorf perfusion of isolated rat hearts. The protein pattern of HS fraction was detected with SDS-PAGE and western blot with ConA and anti ConA antibody. Hearts were then subjected to 20 min ischaemia followed by 20 min reperfusion. During reperfusion, the haemodynamic parameters of hearts were measured. Heart levels of adenine nucleotide were measured in HClO4 extracts using HPLC on C18 column. RESULTS: Liver ischaemia induced changes in protein pattern of HS fraction released from the liver during reperfusion period. Particularly, we registered an increase in amount of several low-molecular weight proteins and decreased amount of high molecular weight proteins. Proteins in this fraction isolated from perfusate after liver ischaemia interact with ConA with lower intensity as proteins isolated from perfusate after control non-ischaemic condition. HS fraction isolated from perfusate after ischaemia and reperfusion of liver had beneficial effect on heart function during 20 min ischaemia and subsequent 20 min reperfusion, documented by: i) decrease of arrhythmia score from 2 to 1 in 5 min of reperfusion and from 2 to 0 in 10 min of reperfusion; ii) improved heart contractility monitored as stabilised [dP/dt]max and increased Q parameter; iii) increased coronary flow. Proteins isolated from liver perfused under control non ischaemic condition did not induce similar effects. The stabilisation of heart haemodynamics, observed after administration of HS proteins isolated from perfusate after ischaemia and reperfusion was associated with slight increase in ATP and ADP levels as well as decrease in AMP level. PMID- 15113130 TI - Unit price and choice in a token-reinforcement context. AB - Pigeons were exposed to multiple and concurrent second-order schedules of token reinforcement, with stimulus lights serving as token reinforcers. Tokens were produced and exchanged for food according to various fixed-ratio schedules, yielding equal and unequal unit prices (responses per unit food delivery). On one schedule (termed the standard schedule), the unit price was held constant across conditions. On a second schedule (the alternative schedule), the unit price was either the same or different from the standard. Under conditions with unequal unit prices, near-exclusive preference for the lower unit price was obtained. Under conditions with equal unit prices, the direction and degree of preference depended on ratio size (number of responses per exchange period). When this ratio differed, strong preferences for the smaller ratio were observed. When this ratio was equal, preferences were nearer indifference. Response rates on the multiple schedule were generally consistent with the preference data in showing sensitivity to ratio size. Results are discussed in terms of a unit-price model that includes handling and reinforcer immediacy as additional costs. On the whole, results show that preferences were determined primarily by delay to the exchange period. PMID- 15113131 TI - Effects of experience on preference between forced and free choice. AB - Preference between forced choice and free choice in concurrent-chain schedules of reinforcement was investigated in pigeons after exposure to particular combinations of terminal links. In Experiment 1, in which terminal links always ended with reinforcers, one of three pairs of terminal links was arranged as preexposure: (a) both terminal links had only one key (forced choice), (b) both terminal links had two keys (free choice), or (c) a combination of forced and free choice was arranged across sessions. In test sessions following the preexposure, pigeons' preferences rapidly shifted to the terminal links with which they had no recent experience. In Experiment 2, the same procedure was repeated except that each terminal link ended intermittently with reinforcers with a probability of .5 and there was no terminal-link arrangement with a combination of free and forced choice. Pigeons showed the same preference changes as in Experiment 1, but the preference changes did not appear immediately at the beginning of test sessions. These data suggest that recent previous experience was a more important determinant of preference than the difference between forced choice and free-choice terminal links. PMID- 15113132 TI - Discounting of delayed food rewards in pigeons and rats: is there a magnitude effect? AB - Temporal discounting refers to the decrease in the present, subjective value of a reward as the time to its receipt increases. Results from humans have shown that a hyperbola-like function describes the form of the discounting function when choices involve hypothetical monetary rewards. In addition, magnitude effects have been reported in which smaller reward amounts are discounted more steeply than larger amounts. The present research examines the cross-species generality of these findings using real rewards, namely food pellets, with both pigeons and rats. As with humans, an adjusting amount procedure was used to estimate the amount of immediate reward judged equal in value to a delayed reward. Different amounts of delayed food rewards (ranging from 5 to 32 pellets in pigeons and from 5 to 20 pellets in rats) were studied at delays varying from 1 s to 32 s. A simple hyperbola, similar to the hyperbola-like mathematical function that describes the discounting of hypothetical monetary rewards in humans, described the discounting of food rewards in both pigeons and rats. These results extend the generality of the mathematical model of discounting. Rates of discounting delayed food rewards were higher for pigeons than for rats. Unlike humans, however, neither pigeons nor rats showed a reliable magnitude effect: Rate of discounting did not vary systematically as a function of the amount of the delayed reward. PMID- 15113133 TI - The effects of behavioral history on response acquisition with immediate and delayed reinforcement. AB - Effects of prior exposure to the experimental chamber with levers present or absent and variable-time (VT) 60-s water deliveries arranged during one, five, or no 1-hr sessions were examined in rats during a 6-hr response-acquisition session in which presses on one lever produced water delivery immediately or after a 15-s resetting delay, and presses on the other lever canceled scheduled water deliveries. Response acquisition was (a) slower to occur when water deliveries were delayed, (b) most consistent in groups that had received five VT sessions, and (c) impaired by the presence of levers only when there had been five VT sessions and water deliveries were delayed during the acquisition session. PMID- 15113134 TI - Bouts of responding: the relation between bout rate and the rate of variable interval reinforcement. AB - By nose poking a lighted key, rats obtained food pellets on either a variable interval schedule of reinforcement or a schedule that required an average of four additional responses after the end of tile variable-interval component (a tandem variable-interval variable-ratio 4 schedule). With both schedule types, the mean variable interval was varied between blocks of sessions from 16 min to 0.25 min. Total rate of key poking increased similarly as a function of the reinforcer rate for the two schedule types, but response rate was higher with than without the four-response requirement. Analysis of log survivor plots of interresponse times showed that key poking occurred in bouts. The rate of initiating bouts increased as a function of reinforcer rate but was either unaffected or was decreased by adding the four-response requirement. Within-bout response rate was insensitive to reinforcer rate and only inconsistently affected by the four-response requirement. For both kinds of schedule, the ratio of bout time to between-bout pause time was approximately a power function of reinforcer rate, with exponents above and below 1.0. PMID- 15113135 TI - Choice in a variable environment: visit patterns in the dynamics of choice. AB - Molar and molecular views of behavior imply different approaches to data analysis. The molecular view privileges moment-to-moment analyses, whereas the molar view supports analysis of more and less extended activities. In concurrent performance, the molar view supports study of both extended patterns of choice and more local patterns of visiting the choice alternatives. Analysis of the present data illustrated the usefulness of investigating order at various levels of extendedness. Seven different reinforcer ratios were presented within each session, without cues to identify them, and pigeons pecked at two response keys that delivered food on variable-interval schedules. Choice changed rapidly within components as reinforcers were delivered and, following each reinforcer, shifted toward the alternative that produced it. If several reinforcers were delivered consecutively by one alternative, choice favored that alternative, but shifted more slowly with each new reinforcer. A discontinuation of such a series of reinforcers by the delivery of a reinforcer by the other alternative resulted in a large shift of choice toward that alternative. These effects were illuminated by analysis of visits to the two alternatives. Changes in visit length occurred primarily in the first postreinforcer visit to the repeatedly reinforced alternative. All other visits tended to be brief and equal. Performance showed multiple signs of moving in the direction of a fix-and-sample pattern that characterized steady-state performance in earlier experiments with many sessions of maintaining each schedule pair. The analyses of extended and local patterns illustrate the flexibility of a molar view of behavior. PMID- 15113136 TI - An all-in-one dry chemistry immunoassay for the screening of coccidiostat nicarbazin in poultry eggs and liver. AB - An automated immunoassay for the detection of nicarbazin residues in poultry eggs and liver was developed. The assay was based on a novel all-in-one dry chemistry concept and time-resolved fluorometry. The analyte specific antibody was immobilized into a single microtiter well and covered with an insulation layer, on top of which the label was dried in a small volume. The extracted sample was added automatically to the dry microtiter well, and the result was available within 18 min. Due to the rapidity and simplicity, the quantitative immunoassay could also be used as a high throughput screening method. The analytical limit of detection for the assay was calculated as 0.1 ng mL(-)(1) (n = 12) and the functional limit of detection as 3.2 ng g(-)(1) for egg (n = 6) and 11.3 ng g( )(1) for liver (n = 6) samples. The sample recovery varied from 97.3 to 115.6%. Typically, the intra-assay variations were less than 10%, and interassay variations ranged between 8.1 and 13.6%. PMID- 15113137 TI - Application of time-of-flight near infrared spectroscopy for detecting sugar and acid contents in apples. AB - A newly constructed optical measurement system was introduced to nondestructively measure the composition of the inside of an apple by time-of-flight near-infrared spectroscopy (TOF-NIRS). As sugar content increased, optical parameters concerned with time-resolved profile of transmitted pulsed light (the attenuance of peak maxima, At, the time delay of peak maxima, Deltat, and the variation of full width at half-maximum, Deltaw) decreased gradually. When the acid content increased, At and Deltaw increased; however, a significant tendency could not be found for Deltat. At, Deltat, and Deltaw were employed as the explanatory variables for multiple linear regression, principle component regression, and partial least-squares analysis. It was possible to predict both sugar and acid contents in an apple with high precision by TOF-NIRS. Especially, the superiority of TOF-NIRS lied in more precise determination of acid content. PMID- 15113138 TI - Sequential fractionation of grape seeds into oils, polyphenols, and procyanidins via a single system employing CO2-based fluids. AB - Pure supercritical CO(2) was used to remove >95% of the oil from the grape seeds. Subcritical CO(2) modified with methanol was used for the extraction of monomeric polyphenols, whereas pure methanol was used for the extraction of polyphenolic dimers/trimers and procyanidins from grape seed. At optimum conditions, 40% methanol-modified CO(2) removed >79% of catechin and epicatechin from the grape seed. This extract was light yellow in color, and no higher molecular weight procyanidins were detected. Extraction of the same sample after removal of the oils and polyphenols, but now under enhanced solvent extraction conditions using methanol as a solvent, provided a dark red solution shown via electrospray ionization HPLC-MS to contain a relatively high concentration of procyanidins. The uniqueness of the study is attested to by the use of CO(2)-based fluids and the employment of a single instrumental extraction system. PMID- 15113139 TI - Steviol quantification at the picomole level by high-performance liquid chromatography. AB - A simple and highly sensitive reversed-phase high-performance liquid chromatographic method (RP-HPLC) has been developed for the determination of steviol (SV) using dihydroisosteviol (DHISV) as an internal standard (IS). SV and DHISV were derivatized by reaction of the acids with 4-(bromomethyl)-7 methoxycoumarin in an aprotic solvent (DMF or acetone). The resulting ester derivatives were separated on an ODS column (250 x 4.6 mm i.d., 5 microm particle size) using fluorescence detection with excitation at 321 nm and emission at 391 nm. The mobile phase consisted of acetonitrile/water (80:20 v/v) with a flow rate of 1 mL min(-)(1). A linear relationship was observed for concentrations between 0.5 and 50 microg/mL of SV, and the detection limit was 100 pg. For application of this method to samples of beer fortified with stevioside, a simple procedure for extraction of the beer with diethyl ether and derivatization in DMF was applied. Whereas beer samples spiked with SV gave a linear response over the range 0.1-15 microg/mL beer, no SV could be detected in beer samples enriched in stevioside that had been stored for over 3 years. The application of the method to plant samples involved preparation of an acid fraction containing the SV analyte, derivatization, and sample cleanup using small silica columns and thin layer chromatography. A sensitive determination of 594 ng of steviol present in 100 mg of dry plant material was performed with high precision and accuracy. PMID- 15113140 TI - Determination of aluminum(III) in crystallized fruit samples using a multicommutated flow system. AB - A multicommutated flow system was developed for the determination of aluminum in crystallized fruit samples. Spectrophotometric determination is based on the reaction of aluminum with chrome azurol S. The binary sampling technique was implemented to improve mixing conditions and to minimize reagent consumption. Three different working zones were established (0.5-5.0, 5.0-25.0, and 10.0-100 ppm) using the zone sampling approach, allowing us to adapt the extent of the in line dilution. The influence of the chemical and physical parameters on the performance of the system was studied. Detection limits of 0.1, 0.6, and 0.8 ppm were obtained for the lowest, the medium, and the highest dispersion system, respectively. The procedure was applied to the determination of aluminum in crystallized fruit extracts. The results were in agreement with those obtained by the reference flame atomic absorption procedure at a 95% confidence level. Repeatability (RSD) was better than 2.4% in all of the three application zones. PMID- 15113141 TI - Analysis of theaflavins and thearubigins from black tea extract by MALDI-TOF mass spectrometry. AB - Black tea contains two major groups of pigments, theaflavins (TFs) and thearubigins (TRs). TFs contain a bis-flavan substituted 1,2-dihydroxy-3,4 benzotropolone moiety. Unlike the TFs, TRs have not yet been characterized. The chemical structure of the TRs remains a mystery. The present paper reports our effort to study the structure of TFs and TRs using delayed pulsed ion extraction of ions generated via the matrix-assisted laser desorption ionization (MALDI) technique, on line with a Linear time-of-flight (TOF) mass spectrometer. Spectra of standard TFs show not only pseudomolecular ions but also ions resulting from fragmentation. The analysis of MALDI-TOF spectra of black tea fractions shows the structure of some TRs, which are similar to those of TFs because the same loss of mass is observed. PMID- 15113142 TI - Enzyme-linked immunosorbent assays based on rabbit polyclonal and rat monoclonal antibodies against isoproturon. AB - This work describes the production and characterization of rabbit polyclonal antisera (pAb) and rat monoclonal antibodies (mAb) against isoproturon. Coating antigen and enzyme-tracer formats were developed. Standard curves for isoproturon were conducted either in 40 mM phosphate buffered saline (PBS) or in Milli-Q water. PAb 352 together with the best enzyme tracer revealed in the optimized ELISA (enzyme tracer format) a test midpoint of 1.06 +/- 0.34 microg/L (n = 19, standard set up in Milli-Q water) with a detection limit of about 0.1 microg/L. The comparable ELISA with mAb IOC 7E1 had test midpoints of 0.07 +/- 0.04 microg/L (n = 7, standards in Milli-Q water) and 0.11 +/- 0.08 microg/L (n = 33; standards in 40 mM PBS). The limits of detection were about 0.003 and 0.01 microg/L in Milli-Q water and PBS, respectively. Noticeable cross reactivities (CRs) were seen with the major metabolites, namely 4-isopropylaniline, 4 isopropylphenylurea, and 1-(4-isopropylphenyl)-3-methylurea. With pAb 352, these CRs were 5%, 7%, and 31%, respectively, and with mAb IOC 7E1, they were 3%, 5%, and ca. 19%, respectively. All arylurea herbicides had only minor CRs, which ranged from no CR (e.g., chlorosulfuron) to a maximum of 3.3% (chlortoluron). Influences of organic solvents (methanol, ethanol, acetonitrile, and acetone) were evaluated. Both pAb- and mAb-based immunoassays showed the highest tolerance for methanol, up to 5%. Ethanol and acetonitrile could not be used above 2% without an influence on the assays. The same was true for acetone, although tested only in the mAb-based assay. Water samples of different origins and matrices were spiked and analyzed with these pAb and mAb ELISAs. The results demonstrated that these immunoassays are useful screening tools. PMID- 15113143 TI - Assessment of strawberry aroma through solid-phase microextraction-gas chromatography and artificial neuron network methods. Variety classification versus growing years. AB - In a previous work, the SPME-GC-MS method (chemical analysis) coupled with KSOM ANN treatment of the results (statistical algorithm) has proved to be efficient to classify 70 strawberry samples harvested in the same year, through the 17 varieties to which they belonged, in a two-dimensional map. As an extension, the present study confirms that these results were not dependent on the year of strawberry production and discusses what effects were observed between results obtained in different years. Samples of different strawberry varieties were harvested during the three campaigns of 2000, 2001, and 2002 and analyzed independently. The chemical data matrix obtained in each case allowed the verification of the proposal that the same discriminative effect could be obtained independently of the year of production by using maps of different sizes. Therefore, 30 measures obtained from samples of 9 varieties in 2000, 54 measures from 13 varieties in 2001, and 80 measures from 20 varieties in 2002 were correctly classified by using 20, 35, and 56 hexagon maps, respectively. In a second analysis based on the 2002 production, the chemical differences between variety aromatic features were noted through the increasing size of the map used. Finally, results relative to 7 varieties cultivated in 2001 and 2002 and stored under exactly the same conditions were computed together for elaborating a single map. An interesting effect of double classification according to the year and the varieties was observed. PMID- 15113144 TI - New method for the determination of benzoic and sorbic acids in commercial orange juices based on second-order spectrophotometric data generated by a pH gradient flow injection technique. AB - Two widely employed antimicrobials, benzoic and sorbic acids, were simultaneously determined in commercial orange juices employing a combination of a flow injection system with pH gradient generation, diode array spectrophotometric detection, and chemometric processing of the recorded second-order data. Parallel factor analysis and multivariate curve resolution-alternating least-squares were used for obtaining the spectral profiles of sample components and concentration profiles as a function of pH, including provisions for managing rank-deficient data sets. An appropriately designed calibration with a nine-sample set of binary mixtures of standards, coupled to the use of the second-order advantage offered by the applied chemometric techniques, allowed quantitation of the analytes in synthetic test samples and also in commercial orange juices, even in the presence of unmodeled interferents (with relative prediction errors of 8.7% for benzoic acid and 2.5% for sorbic acid). No prior separation or sample pretreatment steps were required. The comparison of results concerning commercial samples with a laborious reference technique yielded satisfactory statistical indicators (recoveries were 99.0% for benzoic acid and 101.4% for sorbic acid). PMID- 15113145 TI - Antimicrobial and antioxidant activities of Melissa officinalis L. (Lamiaceae) essential oil. AB - The present study describes antimicrobial and free radical scavenging capacity (RSC) together with the effects on lipid peroxidation (LP) of Melissa officinalis essential oil. The chemical profile of essential oil was evaluated by the means of gas chromatography-mass spectrometry (GC-MS) and thin-layer chromatography (TLC). RSC was assessed measuring the scavenging activity of essential oil on the 2,2-diphenyl-1-picrylhydrazyl (DPPH(*)) and OH(*) radicals. The effect on LP was evaluated following the activities on Fe(2+)/ascorbate and Fe(2+)/H(2)O(2) systems of induction. The antimicrobial activity was tested against 13 bacterial strains and six fungi. The examined essential oil exhibited very strong RSC, reducing the DPPH radical formation (IC(50) = 7.58 microg/mL) and OH radical generation (IC(50) = 1.74 microg/mL) in a dose-dependent manner. According to the GC-MS and TLC (dot-blot techniques), the most powerful scavenging compounds were monoterpene aldehydes and ketones (neral/geranial, citronellal, isomenthone, and menthone) and mono- and sesquiterpene hydrocarbons (E-caryophyllene). Very strong inhibition of LP, particularly in the Fe(2+)/H(2)O(2) system of induction (94.59% for 2.13 microg/mL), was observed in both cases, also in a dose-dependent manner. The most effective antibacterial activity was expressed on a multiresistant strain of Shigella sonei. A significant rate of antifungal activity was exhibited on Trichophyton species. PMID- 15113146 TI - Antioxidants, low molecular weight carbohydrates, and total antioxidant capacity in strawberries (Fragaria x ananassa): effects of cultivar, ripening, and storage. AB - Four cultivars of strawberries (Senga Sengana, BFr77111, Elsanta, and Honeoye) were studied for their content of antioxidants, total antioxidant capacity, and low molecular weight carbohydrates in relation to harvest year, ripening stage, and cold storage. For ascorbic acid, chlorogenic acid, ellagic acid, and total antioxidative capacity, measured in both water-soluble and water-insoluble extracts, there was a 2-5-fold variation among cultivars. Unripe berries contained lower concentrations of chlorogenic acid and p-coumaric acid and also quercetin and kaempferol compared with riper berries. During cold storage for up to 3 days, relatively few changes in the concentration of the different antioxidants occurred. The concentrations of several investigated parameters were interrelated, for example, for ascorbic acid and water-soluble antioxidant capacity and for ellagic acid and water-insoluble antioxidant capacity. The dominating sugars in strawberries were fructose and glucose, but considerable amounts of sucrose were also present, and their contents varied among cultivars, giving a predicted glycemic index of approximately 81. Verbascose, raffinose, and stachyose were found in only minor amounts. The study shows that the concentration of a number of bioactive compounds in strawberries varied according to cultivar, ripening stage, and storage. This information should make it possible to select strawberries with an optimal content of bioactive compounds. PMID- 15113147 TI - Pigments in green tea leaves (Camellia sinensis) suppress transformation of the aryl hydrocarbon receptor induced by dioxin. AB - Environmental contaminants such as dioxins enter the body mainly through diet and cause various toxicities through transformation of the aryl hydrocarbon receptor (AhR). We previously reported that certain natural flavonoids at the dietary level suppress the AhR transformation induced by 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD). In this study, we identified lutein and chlorophyll a and b from green tea leaves as the novel antagonists for AhR. These active compounds suppressed AhR transformation dose-dependently with the 50% inhibitory concentration (IC(50)) values against 0.1 nM TCDD-induced AhR transformation at 3.2, 5.0, and 5.9 microM, respectively. (-)-Epigallocatechin gallate, which is the most abundant flavonoid in green tea leaves, also showed stronger suppressive effects than did other major tea components, with the IC(50) value of 1.7 microM. Thus, these pigments of green tea leaves have the potential to protect from dioxin toxicity through the suppression of AhR transformation. PMID- 15113148 TI - Ovicidal and adulticidal activity of Eucalyptus globulus leaf oil terpenoids against Pediculus humanus capitis (Anoplura: Pediculidae). AB - The toxic effects of Eucalyptus globulus leaf oil-derived monoterpenoids [1,8 cineole, l-phellandrene, (-)-alpha-pinene, 2-beta-pinene, trans-pinocarveol, gamma-terpinene, and 1-alpha-terpineol] and the known Eucalyptusleaf oil terpenoids (beta-eudesmol and geranyl acetate) on eggs and females of the human head louse, Pediculus humanus capitis, were examined using direct contact and fumigation bioassays and compared with the lethal activity of delta-phenothrin and pyrethrum, two commonly used pediculicides. In a filter paper contact bioassay with female P. h. capitis, the pediculicidal activity was more pronounced with Eucalyptus leaf oil than with either delta-phenothrin or pyrethrum on the basis of LT(50) values (0.125 vs 0.25 mg/cm(2)). 1,8-Cineole was 2.2- and 2.3-fold more toxic than either delta-phenothrin or pyrethrum, respectively. The pediculicidal activities of (-)-alpha-pinene, 2-beta-pinene, and (E)-pinocarveol were comparable to those of delta-phenothrin and pyrethrum. l Phellandrene, gamma-terpinene, and 1-alpha-terpineol were relatively less active than delta-phenothrin and pyrethrum. beta-Eudesmol and geranyl acetate were ineffective. 1-alpha-Terpineol and (E)-pinocaveol were highly effective at 0.5 and 1.0 mg/cm(2), respectively, against P. h. capitis eggs. At 1.0 mg/cm(2), (-) alpha-pinene, 2-beta-pinene, and gamma-terpinene exhibited moderate ovicidal activity, whereas little or no ovicidal activity was observed with the other terpenoids and with delta-phenothrin and pyrethrum. In fumigation tests with female P. h. capitis at 0.25 mg/cm(2), 1,8-cineole, (-)-alpha-pinene, (E) pinocarveol, and 1-alpha-terpineol were more effective in closed cups than in open ones, indicating that the effect of the monoterpenoids was largely due to action in the vapor phase. Neither delta-phenothrin nor pyrethrum exhibited fumigant toxicity. Eucalyptus leaf oil, particularly 1,8-cineole, 1-alpha terpineol, and (E)-pinocaveol, merits further study as potential pediculicides or lead compounds for the control of P. h. capitis. PMID- 15113149 TI - Total cranberry extract versus its phytochemical constituents: antiproliferative and synergistic effects against human tumor cell lines. AB - Cranberries (Vaccinium macrocarpon Ait.) are an excellent dietary source of phytochemicals that include flavonol glycosides, anthocyanins, proanthocyanidins (condensed tannins), and organic and phenolic acids. Using C-18 and Sephadex Lipophilic LH-20 column chromatography, HPLC, and tandem LC-ES/MS, the total cranberry extract (TCE) has been analyzed, quantified, and separated into fractions enriched in sugars, organic acids, total polyphenols, proanthocyanidins, and anthocyanins (39.4, 30.0, 10.6, 5.5, and 1.2% composition, respectively). Using a luminescent ATP cell viability assay, the antiproliferative effects of TCE (200 microg/mL) versus all fractions were evaluated against human oral (KB, CAL27), colon (HT-29, HCT116, SW480, SW620), and prostate (RWPE-1, RWPE-2, 22Rv1) cancer cell lines. The total polyphenol fraction was the most active fraction against all cell lines with 96.1 and 95% inhibition of KB and CAL27 oral cancer cells, respectively. For the colon cancer cells, the antiproliferative activity of this fraction was greater against HCT116 (92.1%) than against HT-29 (61.1%), SW480 (60%), and SW620 (63%). TCE and all fractions showed >/=50% antiproliferative activity against prostate cancer cells with total polyphenols being the most active fraction (RWPE-1, 95%; RWPE-2, 95%; 22Rv1, 99.6%). Cranberry sugars (78.8 microg/mL) did not inhibit the proliferation of any cancer cell lines. The enhanced antiproliferative activity of total polyphenols compared to TCE and its individual phytochemicals suggests synergistic or additive antiproliferative interactions of the anthocyanins, proanthocyanidins, and flavonol glycosides within the cranberry extract. PMID- 15113150 TI - Transepithelial transport of chlorogenic acid, caffeic acid, and their colonic metabolites in intestinal caco-2 cell monolayers. AB - Both chlorogenic and caffeic acids exhibited nonsaturable transport in Caco-2 cells, whereas caffeic acid also showed proton-coupled polarized absorption. Thus, the absorption efficiency of caffeic acid was greater than that of chlorogenic acid. Polarized transport of caffeic acid was inhibited by substrates of MCT such as benzoic and acetic acids. Almost all of the apically loaded chlorogenic and caffeic acid was retained on the apical side, and the transepithelial flux was inversely correlated with the paracellular permeability of Caco-2 cells. These results indicate that transport was mainly via paracellular diffusion, although caffeic acid was absorbed to a lesser extent by the monocarboxylic acid transporter (MCT). Furthermore, m-coumaric acid and 3-(m hydroxyphenyl)propionic acid, the main metabolites of chlorogenic and caffeic acid by colonic microflora, competitively inhibited the transport of fluorescein, a known substrate of MCT. This suggests that their absorption could also be mediated by MCT. These findings have exemplified the physiological importance of MCT-mediated absorption in both phenolic acids per se and their colonic metabolites. PMID- 15113151 TI - Intestinal absorption of p-coumaric and gallic acids in rats after oral administration. AB - Ferulic acid (FA) and p-coumaric acid (CA) are absorbed by the monocarboxylic acid transporter (MCT) in Caco-2 cells, although gallic acid (GA) is not. Therefore, the MCT is selective for certain phenolic acids. Absorption of orally administered CA and GA in rats was studied to obtain serum pharmacokinetic profiles and to investigate their intestinal absorption characteristics in vivo. Rats were administered 100 micromol/kg body weight of CA and GA, and blood was collected from the portal vein and abdominal artery after administration. CA, GA, and their metabolites were quantified with a highly selective and sensitive coulometric detection method using high-performance liquid chromatography electrochemical detection. Ingested CA was rapidly absorbed in the gastrointestinal tract in an intact form. The serum concentration of intact CA in the portal vein peaked 10 min after dosing (C(max) was 165.7 micromol/L). In contrast, GA was slowly absorbed, with a t(max) for intact GA of 60 min and a C(max) of 0.71 micromol/L. The area under the curve for intact CA and GA was calculated from the serum concentration profile in the portal vein to be 2991.3 and 42.6 micromol min L(-)(1), respectively. The relative bioavailability of CA against GA was about 70. This is the first demonstration that absorption efficiency of CA is much higher than that of GA in vivo. The absorption characteristics of CA are clearly different from those of GA. These findings are in good agreement with the results obtained in vitro using a Caco-2 cell system. PMID- 15113152 TI - A Kunitz-type inhibitor of coleopteran proteases, isolated from Adenanthera pavonina L. seeds and its effect on Callosobruchus maculatus. AB - The cowpea weevil Callosobruchus maculatus is one of the major pests of Vigna unguiculata cowpea. Digestion in the cowpea weevil is facilitated by high levels of cysteine and aspartic acid proteinases. Plants synthesize a variety of molecules, including proteinaceous proteinase inhibitors, to defend themselves against attack by insects. In this work, a trypsin inhibitor (ApTI) isolated from Adenanthera pavonina seeds showed activity against papain. The inhibition of papain by ApTI was of the noncompetitive type, with a K(i) of 1 microM. ApTI was highly effective against digestive proteinases from C. maculatus, Acanthoscelides obtectus (bean weevil), and Zabrotes subfasciatus (Mexican bean weevil) and was moderately active against midgut proteinases from the boll weevil Anthonomus grandis and the mealworm Tenebrio molitor. In C. maculates fed an artificial diet containing 0.25% and 0.5% ApTI (w/w), the latter concentration caused 50% mortality and reduced larval weight gain by approximately 40%. The action of ApTI on C. maculatus larvae may involve the inhibition of ApTI-sensitive cysteine proteinases and binding to chitin components of the peritrophic membrane (or equivalent structures) in the weevil midgut. PMID- 15113153 TI - alpha-Glucosidase inhibitory activity of some Sri Lanka plant extracts, one of which, Cassia auriculata, exerts a strong antihyperglycemic effect in rats comparable to the therapeutic drug acarbose. AB - Some Sri Lanka plant stuffs were examined regarding in vitro and in vivo alpha glucosidase (AGH) inhibitory actions. According to the results, water extracts and methanol extracts of dried fruits of Nelli (Phylanthus embelica), methanol extracts of dried flowers of Ranawara (Cassia auriculata), and water extracts of latex of Gammalu (Pterocarpus marsupium) were found to have a potential AGH inhibitory activity. In particular, Ranawara methanol extract showed the strongest AGH inhibitory activity in vitro preferably on maltase giving an IC(50) value of 0.023 mg/mL and inhibited the maltase activity competitively. As a result of single oral administration of Ranawara (C. auriculata) methanol extract in Sprague-Dawley rats, a significant and potent lowering of blood glycemic response toward maltose ingestion was observed at 30 min after dosing of 5 mg/kg, thus, concurrently suppressed insulin activity. The ED(50) of the extract (4.9 mg/kg) clearly indicated that the antihyperglycemic effect was as potent as that of therapeutic drug, acarbose (ED(50) 3.1 mg/kg). PMID- 15113154 TI - Phytosterol composition of hybrid Hibiscus seed oils. AB - The seed oils from fifteen hybrid Hibiscus varieties were analyzed for desmethyl sterol content to identify bioactive compounds that could promote the use of these oils for edible applications. Hibiscusis being developed as a new crop with edible and nutraceutical applications for the component tissues and tissue extracts. Previously, hybrid varieties were developed for ornamental purposes on the basis of flower morphology and color. Currently, the effects of selective breeding on seed oil components are of interest as these represent potential natural products with bioactive properties. In the present study, sterol structures were identified as the corresponding trimethyl silyl ether derivatives obtained from the unsaponifiable fraction of the seed oils. This material contained an average of 32 wt % sterols and exhibited a relative composition of sitosterol, 76.3%; campesterol, 10.3%; stigmasterol, 7.3%; 5-avenasterol, 4.4%; and cholesterol, 0.6%. The content of 5-avenasterol showed statistically significant variation among the hybrid varieties with a range of 1.2-5.8%. PMID- 15113155 TI - Metabolism of thymol and trans-anethole in larvae of Spodoptera litura and Trichoplusia ni (Lepidoptera: Noctuidae). AB - Metabolism of the monoterpenoid thymol and the phenylpropanoid trans-anethole, both constituents of essential oils, was investigated following topical and oral administration of the compounds to the tobacco cutworm (Spodoptera litura) and the cabbage looper (Trichoplusia ni). In both species and irrespective of route, administration of thymol resulted in the excretion of its 3-O-beta-glucoside, whereas trans-anethole was hydroxylated on the side chain methyl group. Both metabolites were isolated and their structures elucidated by interpretation of their mass and NMR spectra. Previous experiments indicated that trans-anethole synergized the toxicity of thymol in S. litura, but analyses of feces indicated that metabolism of thymol was not significantly suppressed when the two compounds were orally coadministered to T. ni larvae. PMID- 15113156 TI - 8S globulin of mungbean [Vigna radiata (L.) Wilczek]: cloning and characterization of its cDNA isoforms, expression in Escherichia coli, purification, and crystallization of the major recombinant 8S isoform. AB - Three isoforms of the cDNA of the major 8S globulin of mungbean, 8Salpha, 8Salpha', and 8Sbeta, were isolated, cloned, and characterized. The cDNA sequences of 8Salpha, 8Salpha', and 8Sbeta had open reading frames of 1362, 1359 or 1362, and 1359 bp, respectively, which code for 454, 453 or 454, and 453 amino acids corresponding to molecular weights of 51 973, 51 627 or 51 758, and 51 779, respectively. Homology in terms of cDNA and amino acid sequences was 91-92% between 8Salpha and 8Salpha', 87% between 8Salpha and 8Sbeta, and 86-88% between 8Salpha' and 8Sbeta. The signal peptide was found to be 1-25, 1-24 or 25, and 1 23 for 8Salpha, 8Salpha', and 8Sbeta, respectively, using the signalP website (Nielsen, H.; Engelbrecht, J.; Brunak, S.; von Heijne, G. Protein Eng. 1997, 10, 1-6). The propeptide was determined to be IVHREN. A single site for glycosylation (N-X-S/T) was observed about 90 amino acids from the C terminus. Homology between mungbean 8S isoforms and other 7-8S proteins ranged from 45 to 68% within members of the legume family and 29 to 34% for crops of different species. The major isoform 8Salpha was expressed in Escherichia coli and purified by successive ammonium sulfate fractionation, hydrophobic interaction, and Mono Q column chromatography. The recombinant 8Salpha, but not the native form, was successfully crystallized producing rhombohedral crystals. PMID- 15113157 TI - Biotechnological production of highly soluble daidzein glycosides using Thermotoga maritima maltosyltransferase. AB - The use of soybean isoflavones in food products is limited due to their low hydrophilicity. To enhance its solubility, the isoflavone daidzin was transglycosylated as a model compound using Thermotoga maritima maltosyltransferase (MTase). Four novel transglycosylation products of daidzin were identified by TLC and MALDI-TOF MS: daidzein 7-O-triglucoside, daidzein 7-O pentaglucoside, daidzein 7-O-heptaglucoside, and daidzein 7-O-nonaglucoside. The major product, daidzein 7-O-triglucoside, was purified by C(18) and gel filtration chromatography, and its molecular structure was determined using UV, IR, MALDI-TOF MS, and NMR. The solubility of daidzein 7-O-triglucoside was 7.5 x 10(4) times that of daidzin, suggesting that the transglycosylation greatly enhanced its water solubility. PMID- 15113158 TI - Statistical correlations of primer thermodynamic stability DeltaG degrees for enhanced flax ISSR-PCR cultivar authentication. AB - We revealed four statistically significant correlations related to inter-simple sequence repeat (ISSR) patterns: (1) between thermodynamic free energy DeltaG degrees of ISSR primer sequence and PCR reamplification intensity (dA(i)), (2) between free energy DeltaG degrees of ISSR primer sequence and PIC coefficient quantifying the polymorphism of ISSR patterns, (3) and (4) between free energy DeltaG degrees of anchor sequence of primer and the number of total, and polymorphic bands in ISSR patterns, respectively. Methodological recommendations for effective ISSR primer design were inferred based on revealed correlations. In particular, free energy of ISSR primer sequence is recommended to be DeltaG degrees > 160 kJ/mol of interaction and free energy of flanking anchor sequence in primer to be around DeltaG degrees = 28 kJ/mol of interaction to produce ISSR patterns displaying maximum polymorphism of flax germplasm. PMID- 15113159 TI - Effects of irradiated phytic acid on antioxidation and color stability in meat models. AB - Lipid oxidation and color stability of meats treated with irradiated phytic acid were investigated during storage for 2 weeks at 4 degrees C. The phytic acid in deionized distilled water (DDW) was degraded by irradiation at 10 and 20 kGy, and the irradiated phytic acid showed a strong antiradical activity. For measuring the antioxidant effects of irradiated phytic acid in food models, beef and pork were prepared with DDW (control), irradiated (10 and 20 kGy) or non-irradiated phytic acid, and ascorbic acid as a model system. Irradiated phytic acid significantly inhibited the lipid oxidation in meats compared to the control and ascorbic acid treated samples during storage (P < 0.05). The redness of the meats treated with phytic acid had a higher value than did the control and ascorbic acid treated samples, but a significant difference was not observed in the samples treated with phytic acid regardless of irradiation treatment. Irradiated phytic acid was also effective in inhibiting the loss of heme iron and metmyoglobin formation during storage. Results indicated that irradiation might be helpful for improving the antioxidant activity of phytic acid in meats. PMID- 15113160 TI - Influence of blending on the content of different compounds in the biological aging of sherry dry wines. AB - Principal components analysis to examine the effect of blending (viz. the mixing and transfer of wine between cask rows in a "criaderas and solera" system) on metabolic activity in flor yeasts during biological aging of sherry dry wines was carried out. The variables used in the analysis were the wine compounds most deeply involved in the flor yeast metabolism, namely ethanol, acetaldehyde, glycerol, acetic acid, and l-proline. The greatest blending effect was found to be on the third and second "criadera", which are the stages where the yeasts show a high metabolic activity. The stages holding the oldest wine (viz. the first criadera and the solera) exhibited no differences before and after blending; therefore, the yeasts have a decreased biological activity in them and physical chemical aging processes seemingly prevail over it. PMID- 15113161 TI - Effect of storage on the essential oil composition of Piper nigrum L. fruits of different ripening states. AB - The qualitative and quantitative composition of the essential oil from black, green, and white pepper was determined by using a simultaneous distillation and extraction micromethod for oil isolation and gas chromatography (GC)/flame ionization detection (FID) and GC/mass spectrometry (MS) analysis techniques. The most abundant compounds in pepper oils were (E)-beta-caryophyllene (1.4-70.4%), limonene (2.9-38.4%), beta-pinene (0.7-25.6%), Delta-3-carene (1.7-19.0%), sabinene (0-12.2%), alpha-pinene (0.3-10.4%), eugenol (0.1-41.0%), terpinen-4-ol (0-13.2%), hedycaryol (0-9.1%), beta-eudesmol (0-9.7%), and caryophyllene oxide (0.1-7.2%). Green pepper corn obtained by a sublimation drying method gave more oil (12.1 mg/g) and a much higher content of monoterpenes (84.2%) in the oil than air-dried green pepper corn (0.8 mg/g and 26.8%, respectively). The oil from ground black pepper contained more monoterpenes and less sesquiterprnes and oxygenated terpenoids as compared to green and white pepper oils. After 1 year of storage of pepper samples in a glass vessel at room temperature, the amount of the oils isolated decreased, the content of terpenes decreased, and the amount of oxygenated terpenoids increased. Differently from other pepper samples, 1 year storage of green pepper corn raised the oil amount more than twice of both drying methods. PMID- 15113162 TI - Fatty acid pattern, oxidation product development, and antioxidant loss in muscle tissue of rainbow trout and Dicentrarchus labrax during growth. AB - The levels of hydrophilic, lipophilic, and enzymatic antioxidants, the oxidative damage to lipids and proteins, and the fatty acid patterns of triglyceride and phospholipid fractions were assayed in fresh muscle tissue of rainbow trouts (Oncorhynchus mykiss) and sea basses (Dicentrarchus labrax) during aging, to investigate the correlation between oxidative stress and aging processes in fish. The present studies suggests that lipid peroxidation and accumulation of oxidized proteins during in vivo aging are most likely to be linked with an age-dependent decline of lipophilic antioxidants (CoQH(2), CoQ, and vitamin E) and vitamin C contents in muscle tissue, whereas fish aging is not linked to a decline in antioxidant enzymes and reduced glutathione levels. Lipophilic antioxidant and vitamin C levels represent a reliable marker of oxidative stress during aging, and their determination might be useful for the assessment of fish age. PMID- 15113163 TI - Cultivar and year-to-year variation of phytosterol content in rye (Secale cereale L.). AB - Intake of phytosterols (and -stanols) has been shown to decrease the level of low density lipoprotein cholesterol and thus protect against development of cardiovascular diseases. Therefore, studies on the cultivar and year-to-year variation in phytosterol content in rye grains have been performed. The phytosterol content and composition of different rye cultivars, grown under identical conditions on the same field in three consecutive years, were analyzed. Both cultivar and year-to-year variation in sterol content were statistically significant (p < 0.0001). The total sterol content varied from 1007 +/- 21 mg/kg in the highest yielding cultivar, Tsulpan 3, to 761 +/- 10 mg/kg in the lowest yielding cultivar (Amando in the 1999 harvest). Because the meteorological conditions varied substantially between the different years, it was possible to deduce the impact of varying weather conditions on phytosterol content in the different cultivars. The studied cultivars had all the lowest phytosterol contents in the dry and warm harvest season of 1999. Although there were statistically significant cultivar and year-to-year variations in the sterol composition (p < 0.0001), these were only between 2 and 4% of the total sterol content. PMID- 15113164 TI - Chemical basis for the antifeedant activity of natural hydroxamic acids and related compounds. AB - Natural hydroxamic acids and related compounds derived from the 1,4-benzoxazin-3 one structure show antifeedant activity against the aphid Rhopalosiphum padi. This antifeeding activity is based on the electrophilic character of the hydroxamic acid function, the opening of the hemiacetal function and the lipophilic character of the molecule. In addition, the antifeedant activity of the aqueous extracts of different tissues of Acanthus mollis (Acanthaceae) was determined. The activity observed is attributed to the presence of 2,4-dihydroxy 1,4-benzoxazin-3-one in the extracts. PMID- 15113165 TI - Residue depletion of tilmicosin in chicken tissues. AB - A high-performance liquid chromatography (HPLC) method with detection at 290 nm was modified and validated for the determination of tilmicosin residues in broiler chicken tissues. The limits of detection (LOD) of the method were 0.01 microg/g for muscle and 0.025 microg/g for liver and kidney. Average recoveries ranged from 80.4 to 88.3%. Relative standard deviation values ranged from 5.2 to 12.1%. Residue depletion of tilmicosin in broiler chickens was examined after dosing over a 5-day period by incorporation of the drug into drinking water at 37.5 and 75.0 mg/L. Tilmicosin concentrations in liver and kidney were highest on day 3 of medication and on day 5 in muscle, in both low- and high-dose groups. The residue levels in both groups were significantly higher in liver than in kidney or muscle. A minimum withdrawal time of 9 days was indicated for residue levels in muscle, liver, and kidney tissues below the maximum residue level (MRL). PMID- 15113166 TI - Soil photolysis in a moisture- and temperature-controlled environment. 2. Insecticides. AB - The photolytic degradations of imidacloprid, carbofuran, diazinon, chlorpyrifos, pyridaben, propoxur, and esfenvalerate were independently compared in both moist (75% field moisture capacity at 0.33 bar) and air-dry microbially viable soils at 5 microg/g. All compounds were applied to sandy soil except for propoxur, which was applied to sandy loam soil. Diazinon was applied to both sandy soil and sandy loam soil. The samples were exposed for up to 360 h, depending on the half-life of the compound. Moisture and temperature were maintained through the use of a specially designed soil photolysis apparatus. Corresponding dark control studies were performed concurrently. With the exception of esfenvalerate, the other compounds exhibited significantly shorter half-lives in moist soils, attributed to the increased hydrolysis and microbial activity of the moist soil. The esfenvalerate metabolism was not first order due to limited mobility in the soil because of its very low water solubility. The overall half-life for esfenvalerate was 740 h, as the percent remaining did not drop below 60%. The imidacloprid half life in irradiated moist soil was 1.8 times shorter than in air-dry soils. However, on dry soil the photodegradation showed poor first-order kinetics after 24 h of exposure. The metabolism of carbofuran and diazinon was highly dependent on soil moisture. Carbofuran exhibited 2.2 times longer half-lives when less moisture was available in the soil. Diazinon in moist sandy soil degraded rapidly, but slowed significantly in irradiated and dark control air-dry sandy soil. Diazinon photolysis on sandy loam soil was not first order, as it attained a constant concentration of 54.9%, attributed to decreased mobility in this soil. Chlorpyrifos photolysis was 30% shorter on moist sand than on air-dry sand. Pyridaben photolyzed rapidly throughout the first 72 h of irradiation but maintained 48% through 168 h. Propoxur metabolism in moist sandy loam soil was not first order and did not degrade below 50% after 360 h of exposure, but the overall half-life was still nearly half of that on irradiated air-dry soil. Three of the compounds showed differences in metabolism patterns during exposure on moist or air-dry soil. Typically, the moist soils produced a more linear decline than that seen in the dry soils, corresponding to the susceptibility of the particular chemical to hydrolysis and/or biodegradation. Four of the eight experiments had shorter half-lives in dark control moist soils than in irradiated dry soils. PMID- 15113167 TI - Application of high-Cu compost to dill and peppermint. AB - A controlled environment experiment was conducted to determine the effect of amending soil with various rates of high-Cu compost (0, 20, 40, and 60% compost/soil by volume) on dill (Anethum graveolens L.) and peppermint (Mentha X piperita L.) yields, on fractionation of Cu and Zn in soils, on elemental composition of soil and tissue, and on the essential oils. The compost contained about 2000 mg kg(-)(1) of Cu. Dill yields were greatest in the 20 or 40% treatments, but peppermint yields were greatest in the 20% treatment. Compost additions increased soil pH and electrical conductivity (EC), HNO(3) extractable soil B, Ca, K, Mg, Mn, P, S, Na, and Pb. Additions of high-Cu compost to soil increased tissue P, S, and Na in both crops and Mn, Mo, and Zn in dill but decreased tissue Ca, Cd, and Fe in both crops and Mn, Mo, and Zn in peppermint, increased Cu in all soil fractions including exchangeable, and increased tissue Cu of dill and peppermint as compared to unamended soil. Addition of 60% of high Cu compost to soil resulted in 760-780 mg kg(-)(1) Cu in the growth medium. Nevertheless, Cu content in both crops reached only 12 mg kg(-)(1) DW in the 60% compost treatment, which is below the toxicity levels for plants and below the upper chronic dietary exposure for animals. The application of high-Cu compost altered chemical composition of dill and peppermint essential oils, but oils were free of Cu, Zn, Cd, Ni, Cr, and Pb. Results from this study suggest that mature composts with concentrations of Cu and Zn of 2008 and 321 mg/kg, respectively, can be used as a soil conditioner without risk for phytotoxicity or risk of increasing the normal range of Cu and Zn in crop tissue. However, the long-term effect of the accumulation of heavy metals in soils following repeated compost applications needs to be carefully considered. PMID- 15113168 TI - Flavor release and perception in hard candy: influence of flavor compound compound interactions. AB - The influence of flavor compound-compound interactions on flavor release properties and flavor perception in hard candy was investigated. Hard candies made with two different modes of binary flavor delivery, (1) L-menthol and 1,8 cineole added as a mixture and (2) L-menthol and 1,8-cineole added separate from one another, were analyzed via breath analysis and sensory time-intensity testing. Single-flavor candy containing only L-menthol or 1,8-cineole was also investigated via breath analysis for comparison. The release rates of both L menthol and 1,8-cineole in the breath were more rapid and at a higher concentration when the compounds were added to hard candy separate from one another in comparison to their addition as a mixture (conventional protocol). Additionally, the time-intensity study indicated a significantly increased flavor intensity (measured as overall cooling) for hard candy made with separate addition of these flavor compounds. In conclusion, the flavor properties of hard candy can be controlled, at least in part, by flavor compound-compound interactions and may be altered by the method of flavor delivery. PMID- 15113169 TI - Flavor release and perception in hard candy: influence of flavor compound-flavor solvent interactions. AB - The release kinetics of l-menthol dissolved in propylene glycol (PG), Miglyol, or 1,8-cineole (two common odorless flavor solvents differing in polarity and a hydrophobic flavor compound) were monitored from a model aqueous system via atmospheric pressure chemical ionization mass spectrometry (APCI-MS). Breath analysis was also conducted via APCI-MS to monitor release of l-menthol from hard candy that used PG and Miglyol for l-menthol incorporation. The quantities of l menthol released when dissolved in PG or Miglyol from the model aqueous system were found to be similar and overall significantly greater in comparison to when dissolved in 1,8-cineole. Analogous results were reported by the breath analysis of hard candy. The release kinetics of l-menthol from PG or Miglyol versus from 1,8-cineole were notably more rapid and higher in quantity. Results from the sensory time-intensity study also indicated that there was no perceived difference in the overall cooling intensity between the two flavor solvent delivery systems (PG and Miglyol). PMID- 15113170 TI - The effect of sulfur fertilizer on glucoraphanin levels in broccoli (B. oleracea L. var. italica) at different growth stages. AB - Three sulfur (S) treatements were imposed by applying gypsum to three broccoli cultivars (Claudia, Marathon, and TB-234) known to differ in glucoraphanin content of mature seeds. The S treatments were control (very low added S), low S (23 kg S ha(-)(1)), and high S (92 kg S ha(-)(1)). The gypsum applications during the early vegetative phase of the three broccoli cultivars increased S uptake and the glucoraphanin content in each plant organ. There were significant genotypic differences for the content of both S and glucoraphanin in all plant organs at different growth stages with gypsum applications. A large increase in S and glucoraphanin content was found in the green heads of broccoli and mature seeds. S present in glucoraphanin accounted for only 4-10% of total S content in broccoli heads. However, S present in glucoraphanin in mature seeds accounted for 40-46% of the total S in the seeds of moderate and high glucoraphanin cultivars (Marathon and TB-234). The partitioning of S into glucoraphanin also increased with gypsum applications. Differences in S uptake, S distribution between organs, and partitioning of S into glucoraphanin largely explained the differences in glucoraphanin content in the green heads and mature seeds for the three broccoli cultivars and three S treatments. PMID- 15113171 TI - Studies of the in vitro intestinal metabolism of isoflavones aid in the identification of their urinary metabolites. AB - Soy isoflavones have recently gained considerable interest due to their possible health benefits. However, detailed studies on the metabolism of isoflavones are lacking. The aims of the investigation presented here were (1) to study the in vitro intestinal metabolism of isoflavones and their hydroxylated analogues 3'-OH daidzein, 6-OH-daidzein, 8-OH-daidzein, and 3'-OH-genistein and (2) to characterize the structures of some earlier identified urinary metabolites of soy isoflavones, for which no authentic reference compounds have been available. Isoflavone standards (1-2 mg) were fermented with human fecal flora (16.7%) for 24 h. Metabolites formed during the fermentation were tentatively identified by interpretation of the mass spectra of trimethylsilylated compounds obtained by GC MS. Compounds having hydroxyl groups at 5-position (i.e., genistein and 3'-OH genistein) were completely converted to metabolites that could not be detected by the methods used in this study. The metabolism of daidzein and its hydroxylated analogues, 3'-OH-daidzein, 6-OH-daidzein, and 8-OH-daidzein, occurred to a much lesser extent. Minor amounts of reduced metabolites (i.e., isoflavanones and alpha-methyldeoxybenzoins) of these compounds were tentatively identified in fermentation extracts. The retention times and the mass spectra of reduced isoflavone metabolites, obtained from in vitro fermentations of pure compounds, were utilized to identify unknown urinary metabolites of soy isoflavones. Four novel isoflavone metabolites were identified in human urine collected after soy supplementation: 3' '-OH-O-desmethylangolensin, 3',4',7-trihydroxyisoflavanone, 4',7,8-trihydroxyisoflavanone, and 4',6,7-trihydroxyisoflavanone. PMID- 15113172 TI - Isoflavone characterization and antioxidant activity of ohio soybeans. AB - Seventeen Ohio soybeans were screened for isoflavone content and antioxidant activity. Isoflavone content was determined by C(18) reversed phase high performance liquid chromatography coupled with a photodiode array detector. Antioxidant activities of soybean extracts were measured using 2,2-diphenyl-1 picryl-hydrazyl (DPPH) free radical and photochemiluminescence (PCL) methods. The highest and lowest total isoflavone contents were 11.75 and 4.20 micromol/g soy, respectively, while the average was 7.12 micromol/g soy. Antioxidant activities of soybean extracts ranged from 7.51 to 12.18 micromol butylated hydroxytoluene (BHT) equivalent/g soy using the DPPH method. Lipid and water soluble antioxidant activities of soybean extracts ranged from 2.40 to 4.44 micromol Trolox equivalent/g soy and from 174.24 to 430.86 micromol ascorbic acid equivalent/g soy, respectively, using the PCL method. PMID- 15113173 TI - Identification and occurrence of tryptamine- and tryptophan-derived tetrahydro beta-carbolines in commercial sausages. AB - The identification and occurrence of tetrahydro-beta-carbolines were studied in different kinds of commercial sausages including cooked, fresh, dry-fermented, and ripened sausages, such as salamis and Spanish chorizo, salchichon, fuet, and morcilla, both smoked and unsmoked. Four compounds were identified in several sausages by high-performance liquid chromatography-mass spectrometry (HPLC-MS): 1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid (1), 1-methyl-1,2,3,4 tetrahydro-beta-carboline-3-carboxylic acid diastereoisomers (2a,b), 1,2,3,4 tetrahydro-beta-carboline (3), and 1-methyl-1,2,3,4-tetrahydro-beta-carboline (4). The latter two (3 and 4) are now reported for the first time in meat products. The presence and occurrence of tetrahydro-beta-carbolines were highly variable depending on each particular sample of sausage, and it did not follow a single specific pattern. The concentration range taken as a sum of the four carbolines varied from undetectable levels to 33 microg/g, with the highest content found in ripened, dry-fermented, and smoked sausages (salami, chorizo, and morcilla) and the lowest in cooked sausages (Frankfurt). Formation of tetrahydro-beta-carbolines might occur during elaboration and the ripening process from a chemical condensation between tryptophan or tryptamine and aldehydes (formaldehyde and acetaldehyde). Smoked samples had higher concentrations of formaldehyde-derived 1,2,3,4-tetrahydro-beta-carboline-3 carboxylic acid (1) and 1,2,3,4-tetrahydro-beta-carboline (tryptoline) (3) than those unsmoked. Also, 1 and 3 were more concentrated in the outer part of the sausage, likely to be in contact with smoke. It is concluded that some dry fermented and/or smoked sausages may be significant dietary sources of tetrahydro beta-carbolines. PMID- 15113174 TI - Antioxidant activity of processed table beets (Beta vulgaris var, conditiva) and green beans (Phaseolus vulgaris L.). AB - It has been shown that thermal processing of tomatoes and sweet corn results in increased antioxidant activities despite the loss of vitamin C. Until now, it is unclear whether this positive effect of thermal processing occurs with all crop produce. Therefore, analysis of a root vegetable (beets) and of a legume (green beans) was undertaken to address this question. Antioxidant activity of beets processed under typical commercial processing conditions remained constant despite an 8% loss of vitamin C, a 60% loss of color, and 30% loss of dietary folate. There was a slight but significant 5% increase in phenolic content of processed beets. In contrast, vitamin C and dietary folate content of green beans remained constant, whereas a 32% reduction in phenolic compounds occurred after typical commercial processing conditions. The antioxidant activity of green beans was reduced by 20%. These findings along with previous works suggest that the effects of thermal processing vary with the respective produce crop type. It also reinforces the concept that optimal health benefits may be achieved when a wide variety of plant foods (fruits, vegetables and whole grains) and preparation methods are incorporated into the diet. PMID- 15113175 TI - Heterogeneity in the fine structure of alkali-extractable arabinoxylans isolated from two rye flours with high and low breadmaking quality and their coexistence with other cell wall components. AB - The alkali extractable (AE) arabinoxylans from two rye flours differing in baking quality were studied following sequential extraction of water-unextractable and starch-free rye flour residue with saturated barium hydroxide solution, water and 1 M sodium hydroxide solution (Ba, BaH, and Na, respectively), and further fractionation of isolated fractions by ammonium sulfate precipitation. (1)H NMR and sugar analyses of AE subfractions provided evidence for the presence of lowly branched arabinoxylans (average arabinose-to-xylose ratio, Ara/Xyl approximately 0.5), containing mainly un- and monosubstituted xylopyranosyl residues (Xylp) in the chain. The proportion of this subfraction decreased from 50% in the Ba fraction to 35 and 17% in the Na and BaH fractions, respectively. Other subfractions, rich in both mono- and disubstituted Xylp, represented arabinoxylan populations with intermediate (Ara/Xyl approximately 0.8) and high substitution degree (Ara/Xyl approximately 1.1). The Ba and Na fractions contained phenolic compounds, whereas they were absent in the BaH fraction. The higher ratio of such phenolic compounds to arabinose (PhC/Ara) found in AE arabinoxylans from rye flour of inferior baking quality was one of the most pronounced differences between arabinoxylan populations from rye flours with high and low baking quality. The arabinoxylans from rye flour of high baking quality present in Ba and Na fractions had slightly higher apparent molecular weights (MWs) when compared to those from rye flour with low baking quality. The arabinoxylans present in the BaH fractions, characterized by the highest MWs, had similar MWs. PMID- 15113176 TI - Dynamic high-resolution 1H and 31P NMR spectroscopy and 1H T2 measurements in postmortem rabbit muscles using slow magic angle spinning. AB - Postmortem changes in rabbit muscle tissue with different glycogen status (normal vs low) were followed continuously from 13 min postmortem until 8 h postmortem and again 20 h postmortem using simultaneous magic angle spinning (1)H and (31)P NMR spectroscopy together with measurement of the transverse relaxation time, T(2), of the muscle water. The (1)H metabolite spectra were measured using the phase-altered spinning sidebands (PASS) technique at a spinning rate of 40 Hz. pH values calculated from the (31)P NMR spectra using the chemical shifts of the C-6 line of histidine in the (1)H spectra and the chemical shifts of inorganic phosphate in the (31)P spectra confirmed the different muscle glycogen status in the tissues. High-resolution (1)H spectra obtained from the PASS technique revealed the presence of a new resonance line at approximately 6.8 ppm during the postmortem period, which were absent in muscles with low muscle glycogen content. This new resonance line may originate from the aminoprotons in creatine, and its appearance may be a result of a pH effect on the exchange rate between the amino and the water protons and thereby the NMR visibility. Alternatively, the new resonance line may originate from the aromatic protons in tyrosine, and its appearance may be a result of a pH-induced protein unfolding exposing hydrophobic amino acid residues to the aqueous environment. Further studies are needed to evaluate these hypotheses. Finally, distributed analysis of the water T(2) relaxation data revealed three relaxation populations and an increase in the population believed to reflect extramyofibrillar water through the postmortem period. This increase was significantly reduced (p < 0.0001) in samples from animals with low muscle glycogen content, indicating that the pH is controlling the extent of postmortem expulsion of water from myofibrillar structures. The significance of the postmortem increase in the amount extramyofibrillar water on the water-holding capacity was verified by centrifugation, which showed a reduced centrifugation loss in muscles with low preslaughter glycogen status (0.9 vs 1.9%, p = 0.07). PMID- 15113177 TI - Human fecal metabolism of soyasaponin I. AB - The metabolism of soyasaponin I (3-O-[alpha-L-rhamnopyranosyl-beta-D galactopyranosyl-beta-D-glucuronopyranosyl]olean-12-ene-3beta,22beta,24-triol) by human fecal microorganisms was investigated. Fresh feces were collected from 15 healthy women and incubated anaerobically with 10 mmol soyasaponin I/g feces at 37 degrees C for 48 h. The disappearance of soyasaponin I in this in vitro fermentation system displayed apparent first-order rate loss kinetics. Two distinct soyasaponin I degradation phenotypes were observed among the subjects: rapid soyasaponin degraders with a rate constant k = 0.24 +/- 0.04 h(-)(1) and slow degraders with a k = 0.07 +/- 0.02 h(-)(1). There were no significant differences in the body mass index, fecal moisture, gut transit time, and soy consumption frequency between the two soyasaponin degradation phenotypes. Two primary gut microbial metabolites of soyasaponin I were identified as soyasaponin III (3-O-[beta-D-galactopyranosyl-beta-D-glucuronopyranosyl]olean-12-ene 3beta,22beta,24-triol) and soyasapogenol B (olean-12-ene-3beta,22beta,24-triol) by NMR and electrospray ionized mass spectroscopy. Soyasaponin III appeared within the first 24 h and disappeared by 48 h. Soyasapogenol B seemed to be the final metabolic product during the 48 h anaerobic incubation. These results indicate that dietary soyasaponins can be metabolized by human gut microorganisms. The sugar moieties of soyasaponins seem to be hydrolyzed sequentially to yield smaller and more hydrophobic metabolites. PMID- 15113178 TI - Inactivation kinetics of purified tomato polygalacturonase by thermal and high pressure processing. AB - Tomato polygalacturonase (PG) was extracted from ripe tomatoes and purified by cation exchange and gel filtration chromatography. Cation exchange chromatography yielded two peaks with PG activity: the first peak was identified as PG2 (the heat labile form) and the second one as PG1 (the heat stable form). Both PG2 and PG1 presented a molar mass of 42 kDa when analyzed by SDS-PAGE and an isoelectric point >9.3. Thermal inactivation of purified tomato PG2, at pH 4.4, in the temperature range from 53 to 63 degrees C, followed first-order kinetics. Combined pressure-temperature inactivation of tomato PG2 was studied at 5-55 degrees C/100-600MPa. Under all pressure-temperature conditions, PG2 inactivation followed first-order kinetics. Purified tomato PG1, although more thermostable than PG2, showed a pressure stability very similar to that of PG2. These results indicate that high-pressure processing is an efficient alternative to inactivate tomato PG without the need for applying high temperatures. PMID- 15113179 TI - Immunoenhancing effects of bovine glycomacropeptide and its derivatives on the proliferative response and phagocytic activities of human macrophagelike cells, U937. AB - The immunomodulatory effect of GMP and its derivates on the cell proliferative response of human macrophagelike cell, U937, and its effect on phagocytic activities via incorporation of fluorescence beads were studied. GMP was found to be a potent immunoenhancer at low concentrations, significantly enhancing the proliferation and phagocytic activities of U937. The modulatory function could be radically altered by enzymatic treatments. Pepsin digestion significantly enhanced the degree of cell proliferation and phagocytic activities, whereas trypsin had no significant effect. The immunoenhancing effects decreased significantly after sialidase treatment; however, more than 70% of activity was retained after treatment. GMP with different carbohydrate chains was shown to possess different modulatory capabilities. Sialic acid-rich GMP fractions showed an enhanced response. These findings indicate that both the carbohydrate chains compositions, including the terminal sialic acids and the polypeptide portions of GMP, are essential for the stimulatory effects of GMP on cell proliferation and phagocytic activities of U937. PMID- 15113180 TI - Safety assessment by in vitro digestibility and allergenicity of genetically modified maize with an amaranth 11S globulin. AB - Prospective testing for allergenicity of proteins obtained from sources with no prior history of causing allergy has been difficult to perform. Thus, the objective of this work was to assess the food safety of genetically modified maize with an amaranth globulin protein termed amarantin. Transgenic maize lines evaluated showed, in relation to nontransgenic, 4-35% more protein and 0-44% higher contents of specific essential amino acids. Individual sequence analysis with known amino acid sequences, reported as allergens, showed that none of these IgE elicitors were identified in amarantin. Amarantin was digested within the first 15 min by Simulated Gastric Fluid treatment as observed by Western blot. Expressed amarantin did not induce important levels of specific IgE antibodies in BALB/c mice, as analyzed by ELISA. We conclude that the transgenic maize with amarantin is not an important allergenicity inducer, just as nontransgenic maize. PMID- 15113181 TI - Nutritional study of copper and zinc in grapes and commercial grape juices from Spain. AB - This paper presents the levels of copper and zinc determined in a total of 66 samples of the most widely consumed varieties of white and red grapes in Spain, as well as those of 60 samples of grape juice (39 from white varieties and 21 from red ones) chosen from the main commercial brands in the country. Atomic absorption spectroscopy was used as analytical technique, with electrothermal atomization after digestion of the sample with HNO(3)-H(2)O(2) for grapes and with HNO(3) for grape juice. The mean Zn contents obtained (0.0462 mg/100 g in grapes and 0.0460 mg/100 mL in grape juice) are lower than those provided by most of the more commonly used food composition tables. The mean Cu contents were 0.0515 mg/100 g in grapes and 0.0063 mg/100 mL in grape juice. On the basis of these data and the official data on consumption of grapes and grape juice in Spain, the contribution of both products to the recommended daily intake of zinc (15 and 12 mg/day for healthy adult men and women, respectively) is estimated to be approximately 0.1%, whereas for Cu, this supply represents rather more than 0.25% of the established ESADDI (1.5-3 mg/day in adults). The growing popularity of these products in recent years, on the basis of its nutritional properties and beneficial effects, requires additional data, and the present findings are of potential use to food composition tables. PMID- 15113182 TI - Fungal immunomodulatory protein from Flammulina velutipes induces interferon gamma production through p38 mitogen-activated protein kinase signaling pathway. AB - FIP-fve is a fungal immunomodulatory protein purified from Flammulina velutipes, an edible golden needle mushroom thought to possess potent immunomodulatory properties. When examined for its effects on lymphocytes, FIP-fve exhibited potent mitogenic effects on human peripheral blood lymphocytes, inducing G1/G0 to S phase proliferation. T cells activated by FIP-fve show significant production and secretion of interferon-gamma (IFN-gamma) associated with intercellular adhesion molecule 1 expression but low detectable levels of interleukin-4 in vitro or in vivo. However, SB203580, the p38 mitogen-activated protein kinase (p38 MAPK) inhibitor, can fully abolish the production of IFN-gamma induced by FIP-fve. At the same time, SB203580 only partially prevents the lymphocytes from progressing from G1 to S phase of the cell cycle. These findings demonstrate that FIP-fve is a potent T-cell activator, mediating its effects via cytokine regulation of p38 MAPK. The immunoprophylatic effects of FIP-fve in Th2-mediated allergic anaphylaxis are believed to be associated with the ability of FIP-fve to enhance activation of IFN-gamma-releasing Th1 cells. PMID- 15113183 TI - Compositional equivalency of Cry1F corn event TC6275 and conventional corn (Zea mays L.). AB - Maize (Zea mays L.) plants have been transformed to express a Cry1F insecticidal crystal protein originally isolated from Bacillus thuringiensis Berliner. This protein controls lepidopteran pests of maize, including the European corn borer, Ostrinia nubilalis (Hubner). As part of the safety assessment for crops containing transgenes, a compositional analysis of the food and feed is conducted. This analysis is designed to detect unintended changes in the nutrient and antinutrient content of the raw commodities produced by the crop due to the insertion of the genes into the genomic DNA of the plant (pleotropic effects). Samples of transgenic and nontransgenic maize forage and grain were collected from six field sites located in the U.S. and Canada. Forage samples were analyzed for proximates and minerals, and grain was further analyzed for fatty acids, amino acids, vitamins, secondary metabolites, and antinutrients. Results demonstrated that maize expressing the Cry1F protein was equivalent to nontransgenic maize with respect to these important components. Comparison of the variability within the nontransgenic and transgenic hybrid, as compared to composition values reported in the literature, suggest that factors other than transgenes may contribute more substantially to the composition of crops. PMID- 15113184 TI - Influence of processing parameters on acrylamide formation during frying of potatoes. AB - Consistent evidence suggests that the probable human carcinogen acrylamide is formed in starch-rich foodstuffs through heat-induced interaction of asparagine and reducing sugars during Maillard browning. However, information regarding the influence of processing parameters on acrylamide formation is scarce. We investigated the impact of temperature, heating time, browning level, and surface to-volume ratio (SVR) on acrylamide generation in fried potatoes. Acrylamide content was determined by liquid chromatography (LC) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). In potato shapes with low SVR, acrylamide content consistently increased with increasing temperature and processing times. By contrast, in shapes with intermediate to high SVR, maximal acrylamide formation occurred at 160-180 degrees C, while higher temperatures or prolonged processing times caused a decrease of acrylamide levels. Moreover, browning levels were not a reliable measure of acrylamide content in large-surface products. PMID- 15113186 TI - Synthesis of a silene from 1,1-dilithiosilole and 2-adamantanone. AB - Reaction of 1,1-dilithio-1-sila-2,3,4,5-tetraphenylsilole with 2-adamantanone produces a 5-silafulvene. This represents a new method for synthesis of silenes, leading to the first example of a silapentafulvene. PMID- 15113187 TI - Antisymmetric exchange in [2Fe-2S]1+ clusters: EPR of the Rieske protein from Thermus thermophilus at pH 14. AB - [2Fe-2S] clusters found in the xanthine oxidase family of proteins exhibit an S = 1/2 EPR feature, called signal II, for which one g-value is significantly above g = 2.0. The g-values of signal II cannot be explained with the standard spin coupling model that has been so successful in describing the g = 1.94 signals of [2Fe-2S] ferredoxins. We have studied the EPR spectra of the Rieske protein from Thermus thermophilus at pH 14 and observed a signal II-type EPR spectrum, with g values at 1.81, 1.94, and 2.14. It is shown that the g-values of signal II can be explained by including an antisymmetric exchange term, d.S1xS2, in the spin Hamiltonian. The presence of this term is sensed by EPR if the isotropic exchange coupling constant J is sufficiently small. For the Rieske protein we determined J = 43 cm-1 which is at least 4 times smaller than the J values reported for [2Fe 2S] clusters that yield standard g = 1.94 signals. PMID- 15113188 TI - Exceptionally small attenuation factors in molecular wires. AB - Molecular-wire behavior, that is, an exceptionally small attenuation factor (beta) of 0.01 +/- 0.005 A-1, has been established in novel C60-based donor acceptor ensembles covalently connected through a series of fully conjugated phenylenevinylene oligomers of precise length and constitution. PMID- 15113189 TI - The structure of a biosynthetic intermediate of pyrroloquinoline quinone (PQQ) and elucidation of the final step of PQQ biosynthesis. AB - Pyrroloquinoline quinone (PQQ) is a tricyclic o-quinone, which serves as a cofactor in several enzyme-catalyzed redox reactions in certain bacteria. PQQ is also important for human health, and its role as a vitamin in mammals has recently been suggested. Although much is known about the function of enzymes that use PQQ as cofactor, relatively little is known about the biosynthesis of this coenzyme. Six gene products in Klebsiella pneumoniae (PqqA-F) are involved in PQQ biosynthesis, and PqqC has been shown to catalyze the last step in the pathway. The chemical structure of the substrate for PqqC has remained elusive and has hampered our understanding of the nature of this reaction. In this report we describe the purification and structure of the substrate as deduced by a number of spectroscopic and chemical methods. The substrate is 3a-(2-amino-2 carboxyethyl)-4,5-dioxo-4,5,6,7,8,9-hexahydroquinoline-7,9-dicarboxylic acid-a fully reduced derivative of PQQ, which has not undergone ring cyclization. These results show that PqqC catalyzes a novel reaction, which involves ring closure and an amazing eight-electron oxidation of the substrate. PMID- 15113190 TI - Synthesis, structure, and reductive elimination chemistry of three-coordinate arylpalladium amido complexes. AB - Four three-coordinate arylpalladium amido complexes with a single hindered phosphine were isolated and structurally characterized. Each possessed a T-shaped geometry. Several of these complexes possessed true three-coordinate structures that lacked any additional coordination by ligand C-H bonds. All of the three coordinate complexes underwent reductive elimination to form the corresponding triarylamine. A comparison of the rate of reaction of the three-coordinate compounds demonstrated that the rate of elimination from the pentaphenylferrocenyl di-tert-butylphosphine complex were the fastest. A comparison of the rates of reactions between three-coordinate and four-coordinate complexes showed that the rates were much faster from the three-coordinate complexes. PMID- 15113191 TI - Selectivity control of carbonylation of methanol to dimethyl oxalate and dimethyl carbonate over gold anode by electrochemical potential. AB - New and unique electrocatalysis of gold for the carbonylation of methanol to dimethyl oxalate (DMO) and dimethyl carbonate (DMC) was found. The selectivity to DMO and DMC could be controlled over gold anode by electrochemical potential, as you like. Drastic changes of gold electrocatalysis was due to changes of the oxidation state of gold, Au0 or Au3+. PMID- 15113192 TI - Simplification of protein NOESY spectra using bioorganic precursor synthesis and NMR spectral editing. AB - A novel method is proposed for the analysis of protein NOEs in solution. In this approach, chemically synthesized precursor compounds for the amino acids valine, leucine, and isoleucine are used for amino acid specific labeling of these hydrophobic residues. The methodology is based on a novel synthetic route to 12C,1H,2H Val, Leu, and Ile side chains selectively labeled with 13CH3 only at the terminal methyl group. In an otherwise 12C,1H labeled protein, discrimination between protons bound to 12C and 13C (or 15N) can be achieved using standard isotope-editing NMR pulse schemes. This strategy significantly relieves problems with spectral overlap through selective observation of interresidue methyl NOEs and will thus be a powerful extension of existing biomolecular NMR methodology. PMID- 15113193 TI - Phosphine catalyzed alpha-arylation of enones and enals using hypervalent bismuth reagents: regiospecific enolate arylation via nucleophilic catalysis. AB - Exposure of enones and enals to 20 mol % tributylphosphine in the presence of triarylbismuth(V) dichlorides results in regiospecific aryl transfer to the alpha position of the enone or enal pronucleophile. These results represent the first examples of enolate arylation under the conditions of nucleophilic catalysis. PMID- 15113194 TI - Cinchona alkaloid-lewis acid catalyst systems for enantioselective ketene aldehyde cycloadditions. AB - Asymmetric cinchona alkaloid-catalyzed acid chloride-aldehyde cyclocondensation (AAC) reactions afford enantioenriched 4-substituted and 3,4-disubstituted beta lactones with near perfect absolute and relative stereocontrol. These reactions are characterized by the operational simplicity derived from using commercially available or easily obtained (one-step) reaction catalysts and in situ ketene generation from acid chlorides. The range of aldehyde substrates that serve as effective AAC substrates include sterically hindered aldehydes such as cyclohexanecarboxaldehyde and pivaldehyde. PMID- 15113195 TI - Homogeneous two-component polycondensation without strict stoichiometric balance via the Tsuji-Trost reaction: remote control of two reaction sites by catalysis. AB - Carothers and Flory established the fundamental principles of homogeneous two component polycondensation reactions by which a variety of commonly used polymers such as polyesters and polyamides (nylons) are synthesized even now. One of the most critical factors to achieve a high degree of polymerization is a strict adherence of stoichiometric balance of the two components. An imbalanced ratio of the two starting materials leads to lower degree of polymerization and, consequently, polymers of poor quality. Here we report the details of a strategy, validated by appropriate experiments, which enables an efficient homogeneous two component polycondensation via palladium-catalyzed allylic substitution (the Tsuji-Trost) reaction without strict stoichiometric balance. PMID- 15113196 TI - Chiral Bronsted acid-catalyzed direct Mannich reactions via electrophilic activation. AB - It was found that the phosphoric acid derivatives of general structure 1 serve as highly effective catalysts for the direct addition of acetyl acetone to N-Boc protected arylimines. The beneficial effects of the 3,3'-bisaryl substituents of the catalysts on the enantioselectivity are greatly appreciated, and thus 1d functions as an excellent catalyst. The Bronsted acid-catalyzed direct Mannich reactions presented herein provide an attractive way to construct beta aminoketones under extremely mild conditions. The stereochemical course of this reaction was established through the synthesis of Boc-(S)-phenylglycine methylester. The transformation thus demonstrated is applicable to a useful method for the synthesis of various phenylglycine derivatives. PMID- 15113197 TI - Chirality exchange from sp3 central chirality to axial chirality: benzannulation of optically active diaryl-2,2-dichlorocyclopropylmethanols to axially chiral alpha-arylnaphthalenes. AB - Chirality exchange benzannulation of optically active (1S)-aryl(aryl')-2,2 dichlorocyclopropylmethanols (>99% ee) using TiCl4 successfully proceeded to give axially chiral (M)-alpha-arylnaphthalenes with excellent levels of stereo induction (>99% ee). This unique transformation involves the single-step chirality exchange from sp3 central chirality to axial chirality, that is, a type of excellent memory effect. PMID- 15113198 TI - Enantioselective O- and N-nitroso aldol synthesis of tin enolates. Isolation of three BINAP-silver complexes and their role in regio- and enantioselectivity. AB - The selective generation of three different silver-BINAP catalysts has been achieved via the proper combination of metal/ligand ratio and/or choice of silver salt. Indeed, the X-ray crystallographic study has been used to gain insight into the structure of the catalyst. After the evaluation of each species, 1:1 (AgX.(R) BINAP) complex was found to be the real active species in the O-selective nitroso aldol reaction. On the other hand, a systematic survey of solvent has shown that the optimal 2:1 (AgX.(R)-BINAP) complex is the effective catalyst in the N selective pathway. Thus, a new method of generation of the silver-BINAP catalyst and the synthetic transformations provide not only new insights into the developing area of catalytic enantioselective nitroso aldol synthesis but also clear guidance for the design of an effective catalyst. PMID- 15113199 TI - Experimental measurement of the strength of a C alpha-H...O bond in a lipid bilayer. AB - Due to the apolar nature of the lipid bilayer, the weak Calpha-H...O H-bond is thought to contribute significantly toward the stability of transmembrane helical bundles such as glycophorin A (GPA). Here for the first time we measured the strength of such a bond, using vibrational frequency shifts of a dimeric and nondimeric variants of GPA containing a Gly CD2 label. Although the resulting estimated bond strength of 0.88 kcal/mol is relatively weak, several such bonds could contribute significantly toward bundle stabilization. PMID- 15113200 TI - A phosphorimidate rearrangement for the facile and selective preparation of allylic amines. AB - Allylic phosphorimidates, readily prepared from the combination of an allylic alcohol, an azide, and a chlorophosphite, undergo [3,3]-rearrangement under thermal conditions to provide single isomers of allylic phosphoramidates. This new rearrangement is tolerant of a range of substitution patterns on the reactants. Treatment of the products of the rearrangement with ethanethiolate followed by acid produces a protected allylic amine. This strategy thus provides an attractive and versatile procedure for the preparation of key synthetic intermediates, allylic amines. PMID- 15113201 TI - Chiral lewis Acid catalysis in nitrile oxide cycloadditions. AB - We describe examples of highly regio- and enantioselective nitrile oxide cycloadditions to unsaturated alkenes using substoichiometric amounts of a chiral Lewis acid. Pyrazolidinones proved to be effective achiral templates in the cycloadditions providing C-adducts typically in >30:1 selectivity and 80-99% ee. To avoid potential problems involving coordination of the Lewis acid by amine bases, we have devised a novel method for the generation of unstable nitrile oxides from hydroximinoyl chlorides using Amberlyst 21 as the base. PMID- 15113202 TI - Solvent polarity effects and limited acid catalysis in rearrangements of model radicals for the methylmalonyl-CoA mutase- and isobutyryl-CoA mutase-catalyzed isomerization reactions. AB - The kinetics of reactions of models for the intermediate radicals formed in the methylmalonyl-CoA mutase- and isobutyryl-CoA mutase-catalyzed rearrangements were studied by laser flash photolysis methods. The aldehyde-containing model analogous to the propanal-3-yl radical reacted via 3-exo cyclization with rate constants that varied with solvent polarity (k in the range 2 x 105 to 1 x 107 s 1). The analogous methyl ketone-containing radical reacted 2 orders of magnitude less rapidly, and the ethylthiocarbonyl-containing radical analogue reacted too slowly for kinetic measurements. No acid catalysis was observed in acetic acid, but the CF3CO2H-complexed radicals reacted 1 order of magnitude faster than the uncomplexed radicals. The results indicate that catalysis of the 3-exo radical cyclizations of the radicals formed in the enzymes by hydrogen bonding to an acid, so-called "partial protonation", is not adequate for acceleration of the reactions to the point of kinetic competence. A dissociative mechanism for the radical rearrangements in nature is considered as an alternative. PMID- 15113203 TI - Conductance titration of single-peptide molecules. AB - We have measured the conductance of single peptides covalently bonded to two Au electrodes via S-Au bonds by repeatedly forming a large number of molecular junctions. The conductance decreases exponentially with the peptide length, with a decay constant of beta = 0.9 +/- 0.1 A-1, suggesting that tunneling is the mechanism of electron transport in the peptides. The conductance of the peptides is sensitive to the solution pH, due to the protonation/deprotonation of the amine and carboxyl groups of the peptides, which provides titration measurements based on single-molecule conductance. PMID- 15113204 TI - Synthesis and structure of 2,5,8-triazido-s-heptazine: an energetic and luminescent precursor to nitrogen-rich carbon nitrides. AB - Derivatized s-triazine (C3N3) precursors have seen significant recent use in the production of carbon nitride materials. Larger polycyclic molecular precursors, such as those containing an s-heptazine core (C6N7 or tri-s-triazine), may improve stability and order in carbon nitride products. In this Communication, we describe the synthesis and crystal structure of 2,5,8-triazido-s-heptazine (2). Synthesis of 2 was achieved from melon, an oligomeric s-heptazine synthesized by the pyrolysis of NH4SCN. Melon was converted to molecular 2,5,8-trichloro-s heptazine, which was then transformed to the triazide upon reaction with (CH3)3SiN3. The crystal structure of 2 verifies that the s-heptazine is planar and the azides adopt a pinwheel-like C3h arrangement around the periphery. The s heptazine core shows pi delocalization in the C-N bonds around the periphery (av. 1.33 A), while the internal planar C-N bonds are longer (1.40 A). The heptazine units pack into parallel, but offset, layered sheets in the crystal. The triazide 2 exhibits photoluminescence at 430 nm and rapidly and exothermically decomposes upon heating at 185 degrees C to produce a tan thermally stable carbon nitride powder with a formula near C3N4. PMID- 15113205 TI - Arginine deiminase uses an active-site cysteine in nucleophilic catalysis of L arginine hydrolysis. AB - Arginine deiminase (EC 3.5.3.6) catalyzes the hydrolysis of l-arginine to citrulline and ammonium ion, which is the first step of the microbial l-arginine degradation pathway. The deiminase conserves the active-site Cys-His-Asp motif found in several related enzymes that catalyze group-transfer reactions from the guanidinium center of arginine-containing substrates. For each of these enzymes, nucleophilic catalysis by the conserved Cys has been postulated but never tested. In this communication we report the results from rapid quench studies of single turnover reactions carried out with recombinant Pseudomonas aeruginosa arginine deiminase and limiting [14C-1]l-arginine. The citrulline-formation and arginine decay curves measured at 25 degrees C were fitted to yield apparent rate constants k = 3.6 +/- 0.1 s-1 and k = 4.2 +/- 0.1 s-1, respectively. The time course for the formation (k =13 s-1) and decay (k = 6.5 s-1) of 14C-labeled enzyme defined a kinetically competent intermediate. Under the same reaction conditions, the Cys406Ser mutant failed to form the 14C-labeled enzyme intermediate. These results, along with the recently reported enzyme X-ray structure (Galkin, A.; Kulakova, L.; Sarikaya, E.; Lim, K.; Howard, A.; Herzberg, O. J. Biol. Chem. 2004, 279, 14001-14008, evidence a reaction pathway in which l arginine deimination proceeds via a covalent enzyme intermediate formed by ammonia displacement from the arginine guanidinum carbon by the active-site Cys406. PMID- 15113206 TI - Electrically forced coaxial nanojets for one-step hollow nanofiber design. AB - The outer liquid of a two-liquid coaxial electrified jet is gelled before the onset of natural instabilities to yield hollow nanofibers. By using sol-gel chemistry, innocuous solvents such as glycerol and olive oil, and electrohydrodynamics, it is possible to make such structures in a rather straightforward manner. PMID- 15113207 TI - Spectroscopic and quantum chemical characterization of the electronic structure and bonding in a non-heme FeIV[double bond]O complex. AB - High valent FeIV=O species are key intermediates in the catalytic cycles of many mononuclear non-heme iron enzymes involving the binding and activation of dioxygen. Using variable temperature magnetic circular dichroism (VT MCD) spectroscopy and experimentally calibrated density functional calculations, we are able to present the first detailed description of the electronic structure of a non-heme FeIV=O S = 1 complex. These studies define the nature of the FeIV=O bond and present the basis for understanding high-valent oxygen intermediates in non-heme iron enzymes. PMID- 15113208 TI - Singlet oxygen generation via two-photon excited FRET. AB - A new approach to two-photon excited photodynamic therapy has been developed. A dendritic array of eight donor chromophores capable of two-photon absorption (TPA) was covalently attached to a central porphyrin acceptor. Steady-state fluorescence measurements demonstrated that the donor chromophores transfer excited-state energy to the porphyrin with 97% efficiency. Two-photon excitation of the donor chromophores at 780 nm resulted in a dramatic increase in porphyrin fluorescence relative to a porphyrin model compound. Enhanced singlet oxygen luminescence was observed from oxygen-saturated solutions of the target compound under two-photon excitation conditions. PMID- 15113209 TI - Carbon monoxide induced decomposition of the active site [Ni-4Fe-5S] cluster of CO dehydrogenase. AB - During the past two years, crystal structures of Cu- and Mo-containing carbon monoxide dehydrogenases (CODHs) and Ni- and Fe-containing CODHs have been reported. The active site of CODHs from anaerobic bacteria (cluster C) is composed of Ni, Fe, and S for which crystallographic studies of the enzymes from Carboxydothermus hydrogenoformans, Rhodospirillum rubrum, and Moorella thermoaceticarevealed structural similarities in the overall protein fold but showed substantial differences in the essential Ni coordination environment. The [Ni-4Fe-5S] cluster C in the fully catalytically competent dithionite-reduced CODH II from C. hydrogenoformans (CODHII(Ch)) at 1.6 A resolution contains a characteristic mu(2)-sulfido ligand between Ni and Fe1, resulting in a square planar ligand arrangement with four S-ligands at the Ni ion. In contrast, the [Ni 4Fe-4S] clusters C in CO-treated CODH from R. rubrum resolved at 2.8 A and in CO treated acetyl-CoA synthase/CODH complex from M. thermoacetica at 2.2 and 1.9 A resolution, respectively, do not contain the mu(2)-sulfido ligand between Ni and Fe1 and display dissimilar geometries at the Ni ion. The [Ni-4Fe-4S] cluster is composed of a cubane [Ni-3Fe-4S] cluster linked to a mononuclear Fe site. The described coordination geometries of the Ni ion in the [Ni-4Fe-4S] cluster of R. rubrum and M. thermoacetica deviate from the square-planar ligand geometry in the [Ni-4Fe-5S] cluster C of CODHII(Ch). In addition, the latter was converted into a [Ni-4Fe-4S] cluster under specific conditions. The objective of this study was to elucidate the relationship between the structure of cluster C in CODHII(Ch) and the functionality of the protein. We have determined the CO oxidation activity of CODHII(Ch) under different conditions of crystallization, prepared crystals of the enzyme in the presence of dithiothreitol or dithionite as reducing agents under an atmosphere of N(2) or CO, and solved the corresponding structures at 1.1 to 1.6 A resolutions. Fully active CODHII(Ch) obtained after incubation of the enzyme with dithionite under N(2) revealed the [Ni-4Fe-5S] cluster. Short treatment of the enzyme with CO in the presence of dithiothreitol resulted in a catalytically competent CODHII(Ch) with a CO-reduced [Ni-4Fe-5S] cluster, but a prolonged treatment with CO caused the loss of CO-oxidizing activity and revealed a [Ni-4Fe-4S] cluster, which did not contain a mu(2)-S. These data suggest that the [Ni-4Fe-4S] cluster of CODHII(Ch) is an inactivated decomposition product originating from the [Ni-4Fe-5S] cluster. PMID- 15113210 TI - Preparation of protein gradients through the controlled deposition of protein nanoparticle conjugates onto functionalized surfaces. AB - This paper describes a simple method for the preparation and characterization of protein density gradients on solid supports. The method employs colloidal metal nanoparticles as protein carriers and optical tags and is capable of forming linear, exponential, 1D, 2D, and multiprotein gradients of varying slope without expensive or sophisticated surface patterning techniques. Surfaces patterned with proteins using the procedures described within are shown to support cell growth and are thus suitable for studies of protein-cell interactions. PMID- 15113211 TI - Filipin orientation revealed by linear dichroism. Implication for a model of action. AB - The organization of the polyene antibiotic filipin in membranes containing cholesterol is a controversial matter of debate. Two contradictory models exist, one suggesting a parallel and the other perpendicular organization of filipin with respect to the plane of the membrane. UV-vis linear dichroism, ATR-FTIR, and fluorescence anisotropy decay techniques were combined to study the orientation of filipin in model systems of membranes composed of 1-palmitoyl-2-oleoyl-sn glycero-3-phosphocholine (POPC) or 1,2-palmitoyl-sn-glycero-3-phosphocholine (DPPC) with and without cholesterol. Filipin's orientation is determined by the presence/absence of cholesterol when it is inserted in gel crystalline phase model membranes. When cholesterol (33%) is present in DPPC bilayers, filipin stands perpendicular to the membrane surface as expected in "pore-forming" models. At variance, absence of cholesterol leaves filipin in an essentially random organization in the lipidic matrix. In liquid crystalline phase bilayers (POPC) filipin's orientation is perpendicular to the membrane surface even in absence of cholesterol. Thus filipin's activity/organization depends not only on cholesterol presence but also in the lipid phase domain it is inserted in. These findings were combined with spectroscopy and microscopy data in the literature, solving controversial matters of debate. PMID- 15113212 TI - Ni(II).Arg-Gly-His-DNA interactions: investigation into the basis for minor groove binding and recognition. AB - A study of the minor-groove recognition of A/T-rich DNA sites by Ni(II).L-Arg-Gly His and Ni(II).D-Arg-Gly-His was carried out with a fluorescence-based binding assay, one- and two-dimensional (1D and 2D) NMR methodologies, and molecular simulations. Fluorescence displacement titrations revealed that Ni(II).L-Arg-Gly His binds to A/T-rich sequences better than the D-Arg diastereomer, while NMR investigations revealed that both metallopeptides bind to the minor groove of an AATT core region as evidenced by an intermolecular nuclear Overhauser effect (NOE) between each metallopeptide His imidazole C4 proton and the C2 proton of adenine. Results from molecular dynamics simulations of these systems were consistent with the experimental data and indicated that the His imidazole N-H, the N-terminal peptide amine, and Arg side chains of each metallopeptide are major determinants of minor-groove recognition by functioning as H-bond donors to the O2 of thymine residues or N3 of adenine residues. PMID- 15113213 TI - Strategy for the study of paramagnetic proteins with slow electronic relaxation rates by nmr spectroscopy: application to oxidized human [2Fe-2S] ferredoxin. AB - NMR studies of paramagnetic proteins are hampered by the rapid relaxation of nuclei near the paramagnetic center, which prevents the application of conventional methods to investigations of the most interesting regions of such molecules. This problem is particularly acute in systems with slow electronic relaxation rates. We present a strategy that can be used with a protein with slow electronic relaxation to identify and assign resonances from nuclei near the paramagnetic center. Oxidized human [2Fe-2S] ferredoxin (adrenodoxin) was used to test the approach. The strategy involves six steps: (1) NMR signals from (1)H, (13)C, and (15)N nuclei unaffected or minimally affected by paramagnetic effects are assigned by standard multinuclear two- and three-dimensional (2D and 3D) spectroscopic methods with protein samples labeled uniformly with (13)C and (15)N. (2) The very broad, hyperfine-shifted signals from carbons in the residues that ligate the metal center are classified by amino acid and atom type by selective (13)C labeling and one-dimensional (1D) (13)C NMR spectroscopy. (3) Spin systems involving carbons near the paramagnetic center that are broadened but not hyperfine-shifted are elucidated by (13)C[(13)C] constant time correlation spectroscopy (CT-COSY). (4) Signals from amide nitrogens affected by the paramagnetic center are assigned to amino acid type by selective (15)N labeling and 1D (15)N NMR spectroscopy. (5) Sequence-specific assignments of these carbon and nitrogen signals are determined by 1D (13)C[(15)N] difference decoupling experiments. (6) Signals from (1)H nuclei in these spin systems are assigned by paramagnetic-optimized 2D and 3D (1)H[(13)C] experiments. For oxidized human ferredoxin, this strategy led to assignments (to amino acid and atom type) for 88% of the carbons in the [2Fe-2S] cluster-binding loops (residues 43-58 and 89-94). These included complete carbon spin-system assignments for eight of the 22 residues and partial assignments for each of the others. Sequence specific assignments were determined for the backbone (15)N signals from nine of the 22 residues and ambiguous assignments for five of the others. PMID- 15113214 TI - Structural and rate studies of the 1,2-additions of lithium phenylacetylide to lithiated quinazolinones: influence of mixed aggregates on the reaction mechanism. AB - The 1,2-addition of lithium phenylacetylide (PhCCLi) to quinazolinones was investigated using a combination of structural and rate studies. (6)Li, (13)C, and (19)F NMR spectroscopies show that deprotonation of quinazolinones and phenylacetylene in THF/pentane solutions with lithium hexamethyldisilazide affords a mixture of lithium quinazolinide/PhCCLi mixed dimer and mixed tetramer along with PhCCLi dimer. Although the mixed tetramer dominates at high mixed aggregate concentrations and low temperatures used for the structural studies, the mixed dimer is the dominant form at the low total mixed aggregate concentrations, high THF concentrations, and ambient temperatures used to investigate the 1,2-addition. Monitoring the reaction rates using (19)F NMR spectroscopy revealed a first-order dependence on mixed dimer, a zeroth-order dependence on THF, and a half-order dependence on the PhCCLi concentration. The rate law is consistent with the addition of a disolvated PhCCLi monomer to the mixed dimer. Investigation of the 1,2-addition of PhCCLi to an O-protected quinazolinone implicates reaction via trisolvated PhCCLi monomers. PMID- 15113215 TI - Practical procedures for the preparation of N-tert-butyldimethylsilylhydrazones and their use in modified Wolff-Kishner reductions and in the synthesis of vinyl halides and gem-dihalides. AB - In this work we develop practical chemistry for the preparation of N-tert butyldimethylsilylhydrazone (TBSH) derivatives from carbonyl-containing compounds and show that these products serve as superior alternatives to simple hydrazones in Wolff-Kishner-type reduction reactions, in the Barton vinyl iodide preparation, in the synthesis of vinyl bromides, and in the synthesis of gem diiodides, gem-dibromides, and gem-dichlorides. In our new procedure for silyl hydrazone synthesis, aliphatic and aromatic ketones and aldehydes are shown to undergo highly efficient coupling (typically >95% yield) to form the corresponding TBSH derivatives when combined with equimolar amounts of 1,2 bis(tert-butyldimethylsilyl)hydrazine (BTBSH) and a catalytic quantity of scandium trifluoromethanesulfonate (typically, 0.01 mol %), neat, or in solvent. Optimized procedures are provided for the use of TBSH derivatives in a Wolff Kishner-type reduction protocol that proceeds at low temperature (23-100 degrees C) and in a single reaction flask. Similarly, protocols for the use of TBSH derivatives as precursors to vinyl halides and gem-dihalides are described in detail. PMID- 15113216 TI - Fluorescein provides a resonance gate for FRET from conjugated polymers to DNA intercalated dyes. AB - Fluorescence spectra show that excitation of the cationic water-soluble conjugated polymer poly[(1,4-phenylene)-2,7-[9,9-bis(6'-N,N,N-trimethylammonium) hexyl]fluorene diiodide] (1) results in inefficient fluorescence resonance energy transfer (FRET) to ethidium bromide (EB) intercalated within double-stranded DNA (dsDNA). When fluorescein (Fl) is attached to one terminus of the dsDNA, there is efficient FRET from 1 through Fl to EB. The cascading energy-transfer process was examined mechanistically via fluorescence decay kinetics and fluorescence anisotropy measurements. These experiments show that the proximity and conformational freedom of Fl provide a FRET gate to dyes intercalated within DNA which are optically amplified by the properties of the conjugated polymer. The overall process provides a substantial improvement over previous homogeneous conjugated polymer based DNA sensors, namely, in the form of improved selectivity. PMID- 15113217 TI - Origin, nature, and fate of the fluorescent state of the green fluorescent protein chromophore at the CASPT2//CASSCF resolution. AB - Ab initio CASPT2//CASSCF relaxation path computations are employed to determine the intrinsic (e.g., in vacuo) mechanism underlying the rise and decay of the luminescence of the anionic form of the green fluorescent protein (GFP) fluorophore. Production and decay of the fluorescent state occur via a two-mode reaction coordinate. Relaxation along the first (totally symmetric) mode leads to production of the fluorescent state that corresponds to a planar species. The second (out-of-plane) mode controls the fluorescent state decay and mainly corresponds to a barrierless twisting of the fluorophore phenyl moiety. While a "space-saving" hula-twist conical intersection decay channel is found to lie only 5 kcal mol(-1) above the fluorescent state, the direct involvement of a hula twist deformation in the decay is not supported by our data. The above results indicate that the ultrafast fluorescence decay observed for the GFP chromophore in solution is likely to have an intrinsic origin. The possible effects of the GFP protein cavity on the fluorescence lifetime of the investigated chromophore model are discussed. PMID- 15113218 TI - Solvophobic acceleration of Diels-Alder reactions in supercritical carbon dioxide. AB - The rate of the Diels-Alder reaction between N-ethylmaleimide and 9 hydroxymethylanthracene in supercritical carbon dioxide (scCO(2)) was determined by following the disappearance of 9-hydroxymethylanthracene with in situ UV/vis absorption spectroscopy. The reaction conditions were 45-75 degrees C and 90-190 bar, which correspond to fluid densities (based on pure carbon dioxide) ranging between approximately 340 and 730 kg m(-3). The measured reaction rate at low scCO(2) fluid densities was nearly 25x faster than that reported in acetonitrile at the same temperature (45 degrees C). An inverse relationship between reaction rate and fluid density/pressure was observed at all temperatures in scCO(2). The apparent activation volumes were large and positive (350 cm(3) mol(-1)) and only a weak function of reduced temperature. A solvophobic mechanism analogous to those observed in conventional solvents is postulated to describe (a) the rate acceleration observed for this reaction in scCO(2) relative to that in acetonitrile, (b) the observed relationship between reaction rate and pressure/temperature/density, and (c) the large, positive activation volumes. Solubility measurements in scCO(2), rate measurements in conventional solvents, and an empirical correlation are used to support this theory. Our results advance the general understanding of reactivity in supercritical fluids and provide a rationale for selecting reactions which can be accelerated when conducted in scCO(2). PMID- 15113219 TI - Asymmetric total synthesis of dendrobatid alkaloids: preparation of indolizidine 251F and its 3-desmethyl analogue using an intramolecular Schmidt reaction strategy. AB - Total syntheses of alkaloid 251F (1), a natural product detected from the skin extracts of the dendrobatid frog species Minyobates bombetes, and its racemic 3 desmethyl derivative (2) are reported. A Diels-Alder reaction initiated both syntheses and established four consecutive stereogenic centers. Important to the synthesis of 2 was a first-generation ozonolysis/olefination/aldol strategy to convert a [2.2.1] bicyclic acid to the [3.3.0]bicyclooctane diquinane 4b. Further elaboration to an appropriate keto azide allowed for a key intramolecular Schmidt reaction to deliver the tricyclic core of the target molecule. In a second generation approach, a tandem ring-opening/ring-closing metathesis reaction effected an overall [2.2.1] --> [3.3.0] skeletal rearrangement to deliver diquinane 4a. In similar fashion, 4a was manipulated to an appropriate keto azide, and an intramolecular Schmidt reaction generated the core cyclic architecture of 251F. PMID- 15113220 TI - Diffusion of alpha-tocopherol in membrane models: probing the kinetics of vitamin E antioxidant action by fluorescence in real time. AB - The new fluorescent membrane probe Fluorazophore-L, a lipophilic derivative of the azoalkane 2,3-diazabicyclo[2.2.2]oct-2-ene, is employed to study the quenching of alpha-tocopherol (alpha-Toc) by time-resolved fluorescence in the microheterogeneous environments of Triton XR-100 and SDS micelles, as well as POPC liposomes. Fluorazophore-L has a small nonaromatic fluorescent polar headgroup and an exceedingly long-lived fluorescence (e.g., 140 ns in aerated SDS micelles), which is efficiently quenched by alpha-Toc (3.9 x 10(9) M(-1) s(-1) in benzene). Based on solvatochromic effects and the accessibility by water-soluble quenchers, the reactive headgroup of Fluorazophore-L, along with the chromanol group of alpha-Toc, resides at the water-lipid interface, which allows for a diffusion-controlled quenching in the lipidic environments. The quenching experiments represent an immobile or stationary case; that is, interparticle probe or quencher exchange during the excited-state lifetime is insignificant. Different quenching models are used to characterize the dynamics and antioxidant action of alpha-Toc in terms of diffusion coefficients or, where applicable, rate constants. The ideal micellar quenching model is suitable to describe the fluorescence quenching in SDS micelles and affords a pseudo-unimolecular quenching rate constant of 2.4 (+/- 0.4) x 10(7) s(-1) for a single quencher per micelle along with a mean aggregation number of 63 +/- 3. In Triton micelles as well as in unilamellar POPC liposomes, a two-dimensional (lateral) diffusion model is most appropriate. The mutual lateral diffusion coefficient D(L) for alpha-Toc and Fluorazophore-L in POPC liposomes is found to be 1.8 (+/- 0.1) x 10(-7) cm(2) s(-1), about a factor of 2 larger than for mutual diffusion of POPC, but more than 1 order of magnitude lower than a previously reported value. The comparison of the different environments suggests a quenching efficiency in the order benzene >> SDS micelles > Triton micelles > POPC liposomes, in line with expectations from microviscosity. The kinetic measurements provide important benchmark values for the modeling of oxidative stress in membranes and other lipidic assemblies. The special case of small lipidic assemblies (SDS micelles), for which the net antioxidant efficacy of alpha-Toc may be lower than expected on the grounds of its diffusional behavior, is discussed. PMID- 15113221 TI - Biomimetically inspired total synthesis and structure activity relationships of 1 O-methyllateriflorone. 6 pi electrocyclizations in organic synthesis. AB - The total synthesis of 1-O-methyllateriflorone (2) is described. The construction of the cage-like domain of the molecule involved a biomimetic Claisen/Diels-Alder cascade, whereas the novel spiroxalactone framework was generated by an intramolecular Michael reaction within precursor 16a involving the carboxylate residue as the nucleophile. This finding might bear on the biosynthetic pathway by which nature forms lateriflorone. Described herein is also an interesting cascade sequence involving facile 6 pi electrocyclizations which leads to complex benzopyran systems. The biological evaluation of a small library of lateriflorone analogues and related systems establishing the first SAR within this class of compounds is also included. Among the most active compounds against tumor cells are 2, 16b, 56, 58, and 59. PMID- 15113222 TI - Using ligand bite angles to control the hydricity of palladium diphosphine complexes. AB - A series of [Pd(diphosphine)(2)](BF(4))(2) and Pd(diphosphine)(2) complexes have been prepared for which the natural bite angle of the diphosphine ligand varies from 78 degrees to 111 degrees. Structural studies have been completed for 7 of the 10 new complexes described. These structural studies indicate that the dihedral angle between the two planes formed by the two phosphorus atoms of the diphosphine ligands and palladium increases by over 50 degrees as the natural bite angle increases for the [Pd(diphosphine)(2)](BF(4))(2) complexes. The dihedral angle for the Pd(diphosphine)(2) complexes varies less than 10 degrees for the same range of natural bite angles. Equilibrium reactions of the Pd(diphosphine)(2) complexes with protonated bases to form the corresponding [HPd(diphosphine)(2)](+) complexes were used to determine the pK(a) values of the corresponding hydrides. Cyclic voltammetry studies of the [Pd(diphosphine)(2)](BF(4))(2) complexes were used to determine the half-wave potentials of the Pd(II/I) and Pd(I/0) couples. Thermochemical cycles, half-wave potentials, and measured pK(a) values were used to determine both the homolytic ([HPd(diphosphine)(2)](+) --> [Pd(diphosphine)(2)](+) + H*) and the heterolytic ([HPd(diphosphine)(2)](+) --> [Pd(diphosphine)(2)](2+) + H(-)) bond-dissociation free energies, Delta G(H*)* and Delta G(H-)*, respectively. Linear free-energy relationships are observed between pK(a) and the Pd(I/0) couple and between Delta G(H-)* and the Pd(II/I) couple. The measured values for Delta G(H*)* were all 57 kcal/mol, whereas the values of Delta G(H-)* ranged from 43 kcal/mol for [HPd(depe)(2)](+) (where depe is bis(diethylphosphino)ethane) to 70 kcal/mol for [HPd(EtXantphos)(2)](+) (where EtXantphos is 9,9-dimethyl-4,5 bis(diethylphosphino)xanthene). It is estimated that the natural bite angle of the ligand contributes approximately 20 kcal/mol to the observed difference of 27 kcal/mol for Delta G(H-)*. PMID- 15113223 TI - Hydrolysis of uranium(VI) at variable temperatures (10-85 degrees C). AB - The hydrolysis of uranium(VI) in tetraethylammonium perchlorate (0.10 mol dm(-3) at 25 degrees C) was studied at variable temperatures (10-85 degrees C). The hydrolysis constants (*beta(n,m)) and enthalpy of hydrolysis (Delta H(n,m)) for the reaction mUO(2)(2+) + nH(2)O = (UO(2))(m)(OH)(n)((2m-n))+) + nH(+) were determined by titration potentiometry and calorimetry. The hydrolysis constants, *beta(1,1), *beta(2,2), and *beta(5,3), increased by 2-5 orders of magnitude as the temperature was increased from 10 to 85 degrees C. The enthalpies of hydrolysis, Delta H(2,2) and Delta H(5,3), also varied: Delta H(2,2) became more endothermic while Delta H(5,3) became less endothermic as the temperature was increased. The heat capacities of hydrolysis, Delta C(p(2,2)) and Delta C(p(5,3)), were calculated to be (152 +/- 43) J K(-1) mol(-1) and -(229 +/- 34) J K(-1) mol(-1), respectively. UV/Vis absorption spectra supported the trend that hydrolysis of U(VI) was enhanced at elevated temperatures. Time-resolved laser induced fluorescence spectroscopy provided additional information on the hydrolyzed species at different temperatures. Approximation approaches to predict the effect of temperature were tested with the data from this study. PMID- 15113224 TI - Chemoselective hydrogenation catalysts: Pt on mesostructured CeO2 nanoparticles embedded within ultrathin layers of SiO2 binder. AB - Pt on mesostructured CeO(2) nanoparticles embedded within ultrathin layers of highly structured SiO(2) binder shows highest activity reported with 80% selectivity for the chemoselective hydrogenation of crotonaldehyde. Characterization by transmission electron microscopy (TEM), CO adsorption, and X ray photoelectron spectroscopy (XPS) show the presence of small Pt metal particles, preferentially located on CeO(2) (70%) together with the formation of Pt-CeO(2-x) sites at the interface between Pt and CeO(2) (4 nm) nanoparticles. These sites are able to polarize the carbonyl group and facilitate the selective hydrogenation of this with respect to the double bond. PMID- 15113225 TI - The syntheses of carbocations by use of the noble-gas oxidant, [XeOTeF5][Sb(OTeF5)6]: the syntheses and characterization of the CX3+ (X = Cl, Br, OTeF5) and CBr(OTeF5)2+ cations and theoretical studies of CX3+ and BX3 (X = F, Cl, Br, I, OTeF5). AB - The CCl(3)(+) and CBr(3)(+) cations have been synthesized by oxidation of a halide ligand of CCl(4) and CBr(4) at -78 degrees C in SO(2)ClF solvent by use of [XeOTeF(5)][Sb(OTeF(5))(6)]. The CBr(3)(+) cation reacts further with BrOTeF(5) to give CBr(OTeF(5))(2)(+), C(OTeF(5))(3)(+), and Br(2). The [XeOTeF(5)][Sb(OTeF(5))(6)] salt was also found to react with BrOTeF(5) in SO(2)ClF solvent at -78 degrees C to give the Br(OTeF(5))(2)(+) cation. The CCl(3)(+), CBr(3)(+), CBr(OTeF(5))(2)(+), C(OTeF(5))(3)(+), and Br(OTeF(5))(2)(+) cations and C(OTeF(5))(4) have been characterized in SO(2)ClF solution by (13)C and/or (19)F NMR spectroscopy at -78 degrees C. The X-ray crystal structures of the CCl(3)(+), CBr(3)(+), and C(OTeF(5))(3)(+) cations have been determined in [CCl(3)][Sb(OTeF(5))(6)], [CBr(3)][Sb(OTeF(5))(6)].SO(2)ClF, and [C(OTeF(5))(3)][Sb(OTeF(5))(6)].3SO(2)ClF at -173 degrees C. The CCl(3)(+) and CBr(3)(+) salts were stable at room temperature, whereas the CBr(n)(OTeF(5))(3 n)(+) salts were stable at 0 degrees C for several hours. The cations were found to be trigonal planar about carbon, with the CCl(3)(+) and CBr(3)(+) cations showing no significant interactions between their carbon atoms and the fluorine atoms of the Sb(OTeF(5))(6)(-) anions. In contrast, the C(OTeF(5))(3)(+) cation interacts with an oxygen of each of two SO(2)ClF molecules by coordination along the three-fold axis of the cation. The solid-state Raman spectra of the Sb(OTeF(5))(6)(-) salts of CCl(3)(+) and CBr(3)(+) have been obtained and assigned with the aid of electronic structure calculations. The CCl(3)(+) cation displays a well-resolved (35)Cl/(37)Cl isotopic pattern for the symmetric CCl(3) stretch. The energy-minimized geometries, natural charges, and natural bond orders of the CCl(3)(+), CBr(3)(+), CI(3)(+), and C(OTeF(5))(3)(+) cations and of the presently unknown CF(3)(+) cation have been calculated using HF and MP2 methods have been compared with those of the isoelectronic BX(3) molecules (X = F, Cl, Br, I, and OTeF(5)). The (13)C and (11)B chemical shifts for CX(3)(+) (X = Cl, Br, I) and BX(3) (X = F, Cl, Br, I) were calculated by the GIAO method, and their trends were assessed in terms of paramagnetic contributions and spin-orbit coupling. PMID- 15113226 TI - Nonclassical vs classical metal...H3C-C interactions: accurate characterization of a 14-electron ruthenium(II) system by neutron diffraction, database analysis, solution dynamics, and DFT studies. AB - A neutron diffraction study of the complex RuCl(2)[PPh(2)(2,6-Me(2)C(6)H(3))](2) (1) defines the precise nature of the delta agostic interactions between the unsaturated metal center and two o-methyl groups of the xylyl substituents. The CH(3) carbon atoms lie in the RuP(2) equatorial plane with Ru...C distances of 2.637(7) and 2.668(6) A, whereas four short Ru.H distances (from 2.113(11) to 2.507(11) A) indicate that each methyl group interacts with two C-H bonds. A survey of the X-ray structures with beta, gamma, delta, and epsilon M...H(3)C-C moieties (no neutron data have been previously reported) shows a linear correlation between the angle M.C-C and the torsion of the methyl group about the C-C bond. Thus, the agostic interactions span the range between the classical (M...eta(2)-HC) and the nonclassical (M...eta(3)-H(2)C) types. A solution study of 1 shows intramolecular rearrangement of each xylyl substituent that equilibrates the environments of its two ortho CH(3) groups. Activation parameters, evaluated from the analysis of (1)H NMR line shape as a function of temperature, are Delta H(++) = 9.6 +/- 0.2 kcal mol(-1) with Delta S(++) = -15.4 +/- 0.7 eu (CDCl(3)). The related 14-electron complexes RuX(2)[PPh(2)(2,6 Me(2)C(6)H(3))](2) (X = I, 2; NCO, 3), prepared from 1 and NaX, show a similar dynamic process in solution, with the iodo derivative displaying the most hindered rotation of the xylyl group. A DFT optimization of the complex RuCl(2)[PH(2)(2,6-Me(2)C(6)H(3))](2) (1a) reproduces well the nonclassical Ru...eta(3)-H(2)C agostic mode, whereas the classical Ru...eta(2)-HC one corresponds to a transition state 1b, destabilized by 3.4 kcal mol(-1). A similar barrier (ca. 3.8 kcal mol(-1)) is calculated for the xylyl rotation in the further simplified model RuCl(2)[PH(2)(2,6-Me(2)C(6)H(3))][PH(2)CH[double bond]CHCH(3)] (1c), the absence of bulky phenyl substituents being largely responsible for the difference with respect to the experimental value. Finally, the MO analysis addresses the intrinsic stability of the 14-electron complex RuCl(2)(PH(3))(2) and, in agostic complexes, accounts for the different interactions between the methyl group and the metal atom in relation to the length of their interconnecting chain. PMID- 15113227 TI - Si-E (E = N, O, F) bonding in a hexacoordinated silicon complex: new facts from experimental and theoretical charge density studies. AB - The concept of hypervalency in molecules, which hold more than eight valence electrons at the central atom, still is a topic of constant debate. There is general interest in silicon compounds with more than four substituents at the central silicon atom. The dispute, whether this silicon is hypervalent or highly coordinated, is enlightened by the first experimental charge density determination and subsequent topological analysis of three different highly polar Si-E (E = N, O, F) bonds in a hexacoordinated compound. The experiment reveals predominantly ionic bonding and much less covalent contribution than commonly anticipated. For comparison gas-phase ab initio calculations were performed on this compound. The results of the theoretical calculations underline the findings of the experiment. PMID- 15113228 TI - Widely-applicable gold substrate for the study of ultrathin overlayers. AB - Ultrathin films on gold substrates have been the subject of enormous scientific and technological interest. Comprehensive study of such systems requires concomitant application of a variety of complementary characterization techniques. The reliability of the result is frequently hampered by the fact that different characterization methods impose different requirements on the Au substrate, resulting in the need to use different types of Au substrates for different measurements, possibly influencing the overlayer structure. This results in an average, rather than exact, structure determination. Here, we show that 15-nm-thick Au films evaporated at 0.5 A/sec on silanized glass and annealed are semi-transparent, electrically conducting, and morphologically well-defined, showing a smooth, [111] textured surface. Such Au films provide a high-quality, widely applicable and relatively inexpensive platform for ultrathin overlayers, enabling characterization by a wide spectrum of experimental methods, applied to the same substrate. The exceptional qualities and analytical capabilities of such substrates are demonstrated with several different systems: (i) Cu underpotential deposition (upd); (ii) alkanethiol self-assembly; (iii) formation of Au nanoparticle layers; (iv) binding of the chromophore protoporphyrin IX (PPIX) to a monolayer of 11-mercaptoundecanoic acid (MUA). In the latter case it is shown that the use of Cu(2+) ions for binding between the carboxylate groups of PPIX and MUA promotes better organization of the porphyrin layer. PMID- 15113229 TI - Making a molecular wire: charge and spin transport through para-phenylene oligomers. AB - Functional molecular wires are essential for the development of molecular electronics. Charge transport through molecules occurs primarily by means of two mechanisms, coherent superexchange and incoherent charge hopping. Rates of charge transport through molecules in which superexchange dominates decrease approximately exponentially with distance, which precludes using these molecules as effective molecular wires. In contrast, charge transport rates through molecules in which incoherent charge hopping prevails should display nearly distance independent, wirelike behavior. We are now able to determine how each mechanism contributes to the overall charge transport characteristics of a donor bridge-acceptor (D-B-A) system, where D = phenothiazine (PTZ), B = p oligophenylene, and A = perylene-3,4:9,10-bis(dicarboximide) (PDI), by measuring the interaction between two unpaired spins within the system's charge separated state via magnetic field effects on the yield of radical pair and triplet recombination product. PMID- 15113231 TI - Structure determination of the intramolecular charge transfer state in crystalline 4-(diisopropylamino)benzonitrile from picosecond X-ray diffraction. AB - The molecular structure of photoexcited crystalline 4 (diisopropylamino)benzonitrile (DIABN) is determined by time-resolved X-ray diffraction with a time resolution of 70 ps. Spectroscopic results suggest that an ICT state with a lifetime of 3 ns is produced after photoexcitation. According to structural refinement of the X-ray data (powder diffraction), the torsional angle of the diisopropylamino group with respect to the plane of the phenyl ring of DIABN decreases from 14 degrees in the electronic ground state to 10 degrees in the equilibrated ICT state. PMID- 15113230 TI - Reversal of enantioselectivity in the hydroformylation of styrene with [2S,4S BDPP]Pt(SnCl3)Cl at high temperature arises from a change in the enantioselective determining step. AB - Deuterioformylation of styrene catalyzed by [(2S,4S)-BDPP]Pt(SnCl(3))Cl at 39 degrees C gave 3-phenylpropanal (3) and 2-phenylpropanal (2) (n:i = 1.8, 71% ee (S)-2) with deuterium only beta to the aldehyde carbonyl and in the formyl group. Small amounts of deuterium were also found in the internal (2.8%), cis terminal (1.4%), and trans terminal (1.3%) vinyl positions of the recovered styrene. Deuterioformylation of styrene at 98 degrees C gave 3- (3) and 2-phenylpropanal (2) (n:i = 2.3, 10% ee (R)-2) with deuterium both alpha and beta to the aldehyde carbonyl and in the formyl group. Deuterium was also found in the internal (20%), cis terminal (12%), and trans terminal (12%) vinyl positions of the recovered styrene. These deuterioformylation results establish that platinum hydride addition to styrene is largely irreversible at 39 degrees C but reversible at 98 degrees C. Hydroformylation of (E)- and (Z)-beta-deuteriostyrene at 40 degrees C, followed by oxidation of the aldehydes to acids, and subsequent derivitization to the (S)-mandelate esters confirmed that 84% of 2-phenylpropanal (2) arises from platinum hydride addition to the si-face of styrene, while 73% of 3 phenylpropanal (3) arises from platinum hydride addition to the re-face of styrene. At 100 degrees C, the effect of variable H(2) and CO pressure on n:i, % ee, and TOF of hydroformylation of styrene was investigated. The results are consistent with enantioselectivity not being fully determined until the final hydrogenolysis of a platinum acyl intermediate. PMID- 15113232 TI - Exploring the electronic and mechanical properties of protein using conducting atomic force microscopy. AB - In interfacing man-made electronic components with specifically folded biomacromolecules, the perturbative effects of junction structure on any signal generated should be considered. We report herein on the electron-transfer characteristics of the blue copper metalloprotein, azurin, as characterized at a refined level by conducting atomic force microscopy (C-AFM). Specifically, the modulation of current-voltage (I-V) behavior with compressional force has been examined. In the absence of assignable resonant electron tunneling within the confined bias region, from -1 to 1 V, the I-V behavior was analyzed with a modified Simmons formula. To interpret the variation of tunneling barrier height and barrier length obtained by fitting with the modified Simmons formula, an atom packing density model associated with protein mechanical deformation was proposed and simulated by molecular dynamics. The barrier heights determined at the minimum forces necessary for stable electrical contact correlate reasonably well with those estimated from bulk biophysical (electroanalytical and photochemical) experiments previously reported. At higher forces, the tunnel barrier decreases to fall within the range observed with saturated organic systems. Molecular dynamics simulations revealed changes in secondary structure and atomic density of the protein with respect to compression. At low compression, where transport measurements are made, secondary structure is retained, and atomic packing density is observed to increase linearly with force. These predictions, and those made at higher compression, are consistent with both experimentally observed modulations of tunneling barrier height with applied force and the applicability of the atom packing density model of electron tunneling in proteins to molecular level analyses. PMID- 15113233 TI - Multinuclear NMR investigations of the oxygen, water, and hydroxyl environments in sodium hexaniobate. AB - Solid-state (1)H, (17)O MAS NMR, (1)H-(93)Nb TRAPDOR NMR, and (1)H double quantum 2D MAS NMR experiments were used to characterize the oxygen, water, and hydroxyl environments in the monoprotonated hexaniobate material, Na(7)[HNb(6)O(19)].15H(2)O. These solid-state NMR experiments demonstrate that the proton is located on the bridging oxygen of the [Nb(6)O(19)](8-) cluster. The solid-state NMR results also show that the NbOH protons are spatially isolated from similar protons, but undergo proton exchange with the water species located in the crystal lattice. On the basis of double quantum (1)H MAS NMR measurements, it was determined that the water species in the crystal lattice have restricted motional dynamics. Two-dimensional (1)H-(17)O MAS NMR correlation experiments show that these restricted waters are preferentially associated with the bridging oxygen. Solution (17)O NMR experiments show that the hydroxyl proton is also attached to the bridging oxygen for the compound in solution. In addition, solution (17)O NMR kinetic studies for the hexaniobate allowed the measurement of relative oxygen exchange rates between the bridging, terminal, and hydroxyl oxygen and the oxygen of the solvent as a function of pH and temperature. These NMR experiments are some of the first investigations into the proton location, oxygen and proton exchange processes, and water dynamics for a base stable polyoxoniobate material, and they provide insight into the chemistry and reactivity of these materials. PMID- 15113234 TI - Characterization of fluxional hydrogen-bonded complexes of acetic acid and acetate by NMR: geometries and isotope and solvent effects. AB - 1H, (2)H, and (13)C NMR spectra of enriched CH(3)(13)COOH acid without and in the presence of tetra-n-butylammonium acetate have been measured around 110 K using a liquefied Freon mixture CDF(3)/CDF(2)Cl as a solvent, as a function of the deuterium fraction in the mobile proton sites. For comparison, spectra were also taken of the adduct CH(3)(13)COOH.SbCl(5) 1 and of CH(2)Cl(13)COOH under similar conditions, as well as of CH(3)(13)COOH and CH(3)(13)COO(-) dissolved in H(2)O and D(2)O at low and high pH at 298 K. The low temperatures employed allowed us to detect several well-known and novel hydrogen-bonded complexes in the slow hydrogen bond exchange regime and to determine chemical shifts and coupling constants as well as H/D isotope effects on chemical shifts from the fine structure of the corresponding signals. The measurements show that self association of both carboxylic acids in Freon solution gives rise exclusively to the formation of cyclic dimers 2 and 3 exhibiting a rapid degenerate double proton transfer. For the first time, a two-bond coupling of the type (2)J(CH(3)COOH) between a hydrogen-bonded proton and the carboxylic carbon has been observed, which is slightly smaller than half of the value observed for 1. In addition, the (1)H and (2)H chemical shifts of the HH, HD, and the DD isotopologues of 2 and 3 have been determined as well as the corresponding HH/HD/DD isotope effects on the (13)C chemical shifts. Similar "primary", "vicinal", and "secondary" isotope effects were observed for the novel 2:1 complex "dihydrogen triacetate" 5 between acetic acid and acetate. Another novel species is the 3:1 complex "trihydrogen tetraacetate" 6, which was also characterized by a complex degenerate combined hydrogen bond- and proton-transfer process. For comparison, the results obtained previously for hydrogen diacetate 4 and hydrogen maleate 7 are discussed. Using an improved (1)H chemical shift hydrogen bond geometry correlation, the chemical shift data are converted into hydrogen bond geometries. They indicate cooperative hydrogen bonds in the cyclic dimers; i.e., widening of a given hydrogen bond by H/D substitution also widens the other coupled hydrogen bond. By contrast, the hydrogen bonds in 5 are anticooperative. The measurements show that ionization shifts the (13)C signal of the carboxyl group to low field when the group is immersed in water, but to high field when it is embedded in a polar aprotic environment. This finding allows us to understand the unusual ionization shift of aspartate groups in the HIV pepstatin complex observed by Smith, R.; Brereton, I. M.; Chai, R. Y.; Kent, S. B. H. Nature Struct. Biol. 1996, 3, 946. It is demonstrated that the Freon solvents used in this study are better environments for model studies of amino acid interactions than aqueous or protic environments. Finally, a novel correlation of the hydrogen bond geometries with the H/D isotope effects on the (13)C chemical shifts of carboxylic acid groups is proposed, which allows one to estimate the hydrogen bond geometries and protonation states of these groups. It is shown that absence of such an isotope effect is not only compatible with an isolated carboxylate group but also with the presence of a short and strong hydrogen bond. PMID- 15113235 TI - Electron self-exchange kinetics determined by MARY spectroscopy: theory and experiment. AB - The electron self-exchange between a neutral molecule and its charged radical, which is part of a spin-correlated radical ion pair, gives rise to line width effects in the fluorescence-detected MARY (magnetic field effect on reaction yield) spectrum similar to those observed in EPR spectroscopy. An increasing self exchange rate (i.e., a higher concentration of the neutral molecule) leads to broadening and subsequent narrowing of the spectrum. Along with a series of MARY spectra recorded for several systems (the fluorophores pyrene, pyrene-d(10) and N methylcarbazole in combination with 1,2- and 1,4-dicyanobenzene) in various solvents, a theoretical model is developed that describes the spin evolution and the diffusive recombination of the radical pair under the influence of the external magnetic field and electron self-exchange, thereby allowing the simulation of MARY spectra of the systems investigated experimentally. The spin evolution of the radicals in the pair is calculated separately using spin correlation tensors, thereby allowing rigorous quantum mechanical calculations for real spin systems. It is shown that the combination of these simulations with high resolution, low noise experimental spectra makes the MARY technique a novel, quantitative method for the determination of self-exchange rate constants. In comparison to a simple analytical formula which estimates the self-exchange rate constant from the slope of the linear part of a line width vs concentration plot, the simulation method yields more reliable and accurate results. The correctness of the results obtained by the MARY method is proved by a comparison with corresponding data from the well-established EPR line broadening technique. With its less stringent restrictions on radical lifetime and stability, the MARY technique provides an alternative to the classical EPR method, in particular for systems involving short-lived and unstable radicals. PMID- 15113236 TI - Defluorination of perfluoropropene using Cp*2ZrH2 and Cp2ZrHF: a mechanism investigation from a joint experimental-theoretical perspective. AB - Cp*(2)ZrH(2) (1) (Cp* = pentamethylcyclopentadienyl) reacts with perfluoropropene (2) to give Cp*(2)ZrHF (3) and hydrodefluorinated products under very mild conditions. Initial C-F bond activation occurs selectively at the vinylic terminal position of the olefin to exchange fluorine for hydrogen. Subsequent hydrodefluorination leads to the formation of the n-propylhydride complex Cp*(2)ZrH(CH(2)CH(2)CH(3)), which can be cleaved with dihydrogen to give propane and 1. A theoretical study of the reaction of Cp*(2)ZrH(2) (Cp* = cyclopentadienyl) and CF(2)[double bond]CF(CF(3)) has been undertaken. Several mechanisms have been examined in detail using DFT(B3PW91) calculations and are discussed for this H/F exchange: (a) internal olefin insertion/beta-fluoride elimination, (b) external olefin insertion/beta-fluoride elimination, and (c) F/H metathesis from either an inside or outside approach. Of these, the first case is found to be energetically preferred. Selective defluorination at the terminal carbon has been shown to be favored over defluorination at the substituted and allylic carbons. PMID- 15113238 TI - The World Health Organization, the American Journal of Orthopsychiatry, and Third World countries. PMID- 15113239 TI - Cultural orientation, ethnic affiliation, and negative daily occurrences: a multidimensional cross-cultural analysis. AB - The present research focuses on cultural variations in the experience of daily stresses and strains. It simultaneously examines the experiences of daily hassles among people holding different cultural orientations (individualistic vs. collectivist) and different socioethnic groups (Jews and Arabs). Data were gathered from 662 Jewish and 300 Arab Israeli respondents by means of a random telephone number dialing. Differences were found in self-related hassles between individuals holding different cultural orientations and in family-related hassles between members of different ethnic affiliations. Multidimensional scaling analysis revealed a different configuration of relations among hassle domains in four groups of Ethnic Affiliation X Cultural Orientation, yielding a continuum from most typical individualists to most typical collectivists, with groups in cultural transition found in between. PMID- 15113240 TI - What works for survivors of childhood abuse: learning from participants in an inpatient treatment program. AB - Thirty adults were interviewed about their experiences in a 6-week program for recovery from posttraumatic stress. The most valued aspects were their relationships with staff and the staff's expertise in treating the long-term effects of abuse. They valued the structured group treatment that allowed them to process emotions with the goal of healing. A few were upset by process groups, whereas others had difficulty with the sudden loss of support at discharge from the program. PMID- 15113241 TI - A survey of the assessment practices of child and adolescent clinicians. AB - Practicing child and adolescent clinicians (N=309) completed a survey regarding their actual and preferred child and adolescent assessment practices. Most reported using the following tools: family interview, individual child/teen interview, and a review of previous treatment records. The majority reported interviewing the identified child/teen patient and the adults in the home with a parental role. Clinicians reported they would also prefer to use parent rating scales, teacher rating scales, child/teen self-report scales, naturalistic observation, and a review of recent report cards and previous educational testing. The top 3 factors reported to influence assessment practice were ethics, organizational pressures, and theoretical orientation. Twenty-one percent of the sample reported that research findings are unimportant in their assessment decision making. PMID- 15113242 TI - Proactive and reactive aggression in referred children and adolescents. AB - Investigating different types of aggression is important to facilitate a better understanding of excessive maladaptive aggression in referred youth. Using regression analysis, the authors investigated demographic, historical, diagnostic, and treatment correlates of proactive aggression and reactive aggression in a heterogeneous population (N = 323) of psychiatrically referred youths. Ratings of proactive and reactive aggression significantly correlated with more established measures of aggression. Results suggest the importance of hyperactive/impulsive behavior, disruptive behavior disorders, and self-reported hostility in youths with both reactive and proactive aggression. Substance use disorders, a family history of substance abuse, and family violence were specifically associated with proactive aggression. Younger age and a history of abuse were correlated with reactive aggression. Implications for clinical interventions and future research are discussed. PMID- 15113243 TI - Emotional resilience: risk and protective factors for depression among alternative education students in New Zealand. AB - A total of 268 alternative education students from New Zealand were surveyed to examine risk and protective factors for depression within the social environment. Twenty-eight (35.4%) of the girls and 31 (21.1%) of the boys had cut-off scores that indicated a high likelihood of significant psychopathology from depression. Multivariate analysis demonstrated that family and peer connections were protective against depression. High levels of poverty, witnessing violence at home, and experiencing bullying at school were significant risk factors for depression. PMID- 15113244 TI - Adult attachment and sexual offender status. AB - This study investigates whether insecure attachment distinguishes sexual from nonsexual offenders and whether insecure attachment is reflected in the choices of victims, modus operandi, and nature of the sexual acts. Incarcerated male sexual and nonsexual offenders were surveyed. Insecure attachment distinguished sexual offenders from nonsexual offenders but was not related to the characteristics of the sexual offenses, with the exception of victim age. The methodological, theoretical, and clinical implications of the findings are discussed, and an alternative line of inquiry is proposed. PMID- 15113245 TI - Treating complex trauma in women within community mental health. AB - Community mental health clinicians are likely to find their case loads composed of women who have complicated trauma histories. In response to the absence of comprehensive treatment for trauma survivors within the community mental health system, an alternative model, Overcoming Pain and Adversity in Life (OPAL) is offered. As an intensive treatment program, OPAL is structured in a triphase format to accommodate the individual needs of each woman and to promote symptom reduction and/or resolution. PMID- 15113246 TI - Parental arrest and children involved with child welfare services agencies. AB - Data from the National Survey of Child and Adolescent Well-Being show that approximately 1 in 8 (12.5%) children who are subjects of reports of maltreatment investigated by child welfare services (CWS) agencies have parents who were recently arrested. Compared with other children who come to the attention of CWS agencies, those with arrested parents are younger, disproportionately African American, and significantly more likely tp be in out-of-home care. Approximately 2 in 5 children age 2 and older with arrested parents had a clinically significant emotional or behavioral problem, yet only 1 in 10 received mental health care. Although parent characteristics varied by race, rates of substance abuse, serious mental illness, domestic violence, and problems meeting basic needs were higher among arrested parents than among other parents. PMID- 15113247 TI - Accommodative coping and well-being of midlife parents of children with mental health problems or developmental disabilities. AB - This study examined how accommodative coping via flexible goal adjustment affects the wellbeing of midlife parents. Using data from the Wisconsin Longitudinal Study, a population-based study of midlife adults, the authors compared parents who have a child with a severe mental health problem, a child with a developmental disability, or a child with no chronic illness or disability. Overall, parents had better well-being (i.e., lower levels of depressive and physical symptoms, higher levels of environmental mastery and self-acceptance) if they used accommodative coping. This effect was stronger for parents of individuals with a severe mental health problem than for the comparison group. PMID- 15113248 TI - Pathologic adaptation to community violence among inner-city youth. AB - The study tests the thesis of pathologic adaptation for youth exposed to community violence, where high levels of exposure to community violence lead to increased aggressive behavior but decreased psychological distress. Four hundred seventy-one 6th graders and 1 of their parents were interviewed. The results showed, for a small but important subgroup of youth, that high levels of exposure to community violence were associated with more child- and parent-reported aggressive behavior and less child-reported psychological distress. Targeted prevention strategies for these high-risk youth are especially needed. PMID- 15113249 TI - A group for men who abuse their partners: participant perceptions of what was helpful. AB - This exploratory study of 88 self-reports examined the subjective perceptions of men attending a group treatment program to address their violent and abusive behaviors. For the majority of participants, this was a positive experience, with education and self-control described as the most helpful learning experience. No clear themes emerged regarding negative experiences with the group. PMID- 15113250 TI - Sniffing in infant rats during sleep and wakefulness. AB - Sniffing, a behavior that enhances detection and localization of odorants, is typically assumed to require behavioral arousal. In an effort to determine whether sniffing and arousal are dissociable, dimethyl disulfide (DMDS) was presented to 8-day-old rats while respiration and behavioral state were monitored. Pups sniffed in response to the highest concentrations of DMDS, exhibiting a lower olfactory threshold when awake. Surprisingly, sniffing occurred even while pups remained asleep. Sniffing was mediated by the olfactory system, as evidenced by the abolition of sniffing when the lateral olfactory tracts, were cut and the retention of rapid arousal in response to a trigeminal stimulant, acetic acid. Finally, sleeping pups presented with acetic acid awakened without sniffing. Thus, although olfactory threshold increases during sleep, sleeping does not preclude sniffing. PMID- 15113252 TI - Mother-Infant Interactions in Rats Lacking CCK(A) Receptors. AB - Mediation of mother-infant interactions by the brain-gut peptide cholecystokinin (CCK) was examined by observing behavior of Otsuka Long-Evans Tokushima Fatty (OLETF) rats, which lack functional CCKA receptors because of a genetic abnormality. OLETF (n = 10) and control (Long-Evans Tokushima Otsuka [LETO] n = 10) dams interacted with 1 pup of each line on Postpartum Days 6-9. OLETF pups received more body and anogenital licking and emitted substantially more ultrasonic vocalizations than LETO pups. OLETF dams carried pups less frequently and showed a nursing position more frequently than LETO dams. No significant Pup X Dam Line interactions or line differences in dams' activity were detected. The results provide convergent validity to previous pharmacological studies implicating CCK mediation of both infant and maternal behavior. PMID- 15113251 TI - Corticosterone influences on Mammalian neonatal sensitive-period learning. AB - Infant rats exhibit sensitive-period odor learning characterized by olfactory bulb neural changes and odor preference acquisitions critical for survival. This sensitive period is coincident with low endogenous corticosterone (CORT) levels and stress hyporesponsivity. The authors hypothesized that low corticosterone levels modulate sensitive-period learning. They assessed the effects of manipulating CORT levels by increasing and removing CORT during (Postnatal Day 8) and after (Postnatal Day 12) the sensitive period. Results show that (a) exogenous CORT prematurely ends sensitive-period odor-shock-induced preferences; (b) adrenalectomy developmentally extends the sensitive period as indicated by odor-shock-induced odor-preference learning in older pups, whereas CORT replacement can reinstate fear learning; and (c) CORT manipulation modulates olfactory bulb correlates of sensitive-period odor learning in a manner consistent with behavior. PMID- 15113253 TI - Enduring effects of prenatal cocaine exposure on selective attention and reactivity to errors: evidence from an animal model. AB - Adult Long-Evans rats, exposed prenatally to 1 of 4 doses of cocaine (0.0,0.5,1.0, or 3.0 mg/kg iv), were tested on a 3-choice visual attention task with an olfactory distractor presented unpredictably on one third of the trials. The performance of all 3 cocaine-exposed groups was significantly more disrupted than that of controls by the presentation of distractors. Results demonstrate that prenatal cocaine exposure increases susceptibility to distractors, using a task specifically designed to measure this function. In addition, the present study revealed that individuals exposed to cocaine in utero exhibit greater performance disruption after an error than controls, in certain types of tasks. Both areas of dysfunction, impaired selective attention and impaired arousal regulation, have important functional consequences in humans, possibly affecting the school performance and social development of cocaine-exposed children. PMID- 15113254 TI - Neonatal amygdala lesions and juvenile isolation in the rat: differential effects on locomotor and social behavior later in life. AB - Pervasive developmental disorders such as autism are characterized by deficits in social interaction and communication. Disturbed development of limbic structures such as the amygdala might underlie these deficits. The authors examined the effects of amygdala lesions on Postnatal Day 7 and juvenile isolation (2 weeks of individual housing during Weeks 4 and 5 of life) on rat locomotor and social activity later in life. Before puberty, but more pronounced after puberty, lesioned rats displayed enhanced locomotor activity. Adult social behavior was selectively disturbed by the lesion and the isolation procedure. In particular, the combination of neonatal lesions and juvenile isolation severely disrupted social interaction. These results suggest that a combination of neonatal amygdala damage and juvenile isolation may serve as an animal model of certain psychopathological neurodevelopmental disorders, such as autism. PMID- 15113256 TI - Estrogen and/or progesterone administered systemically or to the amygdala can have anxiety-, fear-, and pain-reducing effects in ovariectomized rats. AB - Estrogen (E2) and/or progesterone (P) in the amygdala may influence anxiety, fear, and pain behaviors. Ovariectomized rats were administered subcutaneous or intra-amygdala vehicle, E2, P, or E2 + P: Effects on open field, elevated plus maze, defensive freezing, and hot-plate task performance were observed. Subcutaneous E2 + P or intra-amygdala E2, P, or E2 + P increased open field central entries and open arm time in the plus-maze compared with vehicle. Subcutaneous or intra-amygdala E2, P, or E2 + P decreased time spent freezing postshock compared with vehicle. Subcutaneous or intra-amygdala E2 + P increased latencies to lick paws compared with vehicle. Thus, E2 and P may have effects in the amygdala to decrease anxiety, fear, and/or pain responses. PMID- 15113257 TI - Working and reference memory in seizure-prone and seizure-resistant rats: impact of amygdala kindling. AB - In rat selectively bred for different amygdala kindling rates (Fast vs. Slow), comorbid differences in learning were detected. Here, performance was tested in a delayed alternation task before, during, and after kindling. Although similar reference memory was evident, Fast rats showed working memory deficits with increasing delays between information and choice trials. Further, seizures shortly before learning disrupted both reference and working memory in Fast, but not Slow, rats. Weeks after kindling, progressive delays further disrupted Fast rats, but only longer delays disrupted Slow rats. Clearly relevant to individual differences in human epilepsy, a temporal lobe, seizure-prone genetic background in rats provides poorer original learning and easier disruption of new learning by recent and past seizures than a seizure-resistant background. PMID- 15113258 TI - Medial amygdala modulation of predator odor-induced unconditioned fear in the rat. AB - This study examined the participation of the medial amygdala (MeA) in unconditioned fear. Rats received ibotenic acid lesions in the MeA or central amygdala (CeA) prior to cat-odor exposure. MeA-lesioned rats exhibited a significant reduction in freezing duration and made frequent contact with a cloth containing cat odor. In contrast, CeA lesions had no significant effects on unconditioned fear. The freezing reduction produced by MeA lesions was not due to a performance deficit because MeA-lesioned rats, unlike CeA-lesioned rats, were capable of freezing in postshock test intervals. Furthermore, MeA lesions did not alter olfactory function and general locomotor activity. Results demonstrate that the MeA plays a major role in modulating predator odor-induced unconditioned fear. PMID- 15113259 TI - Lateralization of spatial learning in the avian hippocampal formation. AB - The authors investigated lateralization of spatial learning within the avian hippocampal formation (HF). In Experiment 1, homing pigeons (Columba livia) with unilateral lesions of the right or left HF were trained to locate a goal in a square room containing local landmarks and global room cues. All groups learned the task. During probe trials, when landmarks were rotated or removed, intact pigeons and left HF-lesioned pigeons relied exclusively on global room cues to locate the food goal. Pigeons with right HF lesions were the only group to demonstrably use the landmarks. The results suggest that the right HF is preferentially involved in the representation of global environmental space, whereas only the left HF may be sensitive to local landmarks for navigation. PMID- 15113260 TI - Neuron production, neuron number, and structure size are seasonally stable in the hippocampus of the food-storing black-capped chickadee (Poecile atricapillus). AB - Neuronal recruitment in the black-capped chickadee (Poecile atricapillus) hippocampus occurs at a higher rate in the fall than at other times of the year. As a means of determining whether this increase in recruitment results from greater neuron production, chickadees were caught in the wild between October and March and injected with the cell-birth marker 5-bromo-2'-deoxyuridine. Two weeks later, birds were killed by overdose, and hippocampal neuron production, apoptosis, neuron number, and hippocampal volume were determined. Chickadees collected in October, November, January, February, and March did not differ in neuron production, apoptosis, hippocampal volume, or neuron number. These findings indicate that increases in neuronal recruitment in the chickadee hippocampus in the fall do not result from increased neuron production, but instead, enhanced survival of new neurons. PMID- 15113261 TI - Dopamine antagonists do not block conditioned place preference induced by paced mating behavior in female rats. AB - The authors assessed the behavioral effects of dopamine (DA) receptor antagonists, Cis (Z) flupentixol and S(+)-raclopride L-tartrate, on conditioned place preference (CPP) induced by paced mating behavior. Ovariectomized female rats of the Wistar strain were used. The administration of amphetamine (1 mg/kg) induced a clear CPP that was completely blocked by the DA antagonists flupentixol (0.25 mg/kg) or raclopride (0.125 mg/kg). These doses had no effect on motor coordination. Female rats that mated in a pacing chamber developed a clear CPP. Neither flupentixol nor raclopride blocked the reward state induced by paced mating behavior. These results indicate that DA is not involved in the reward state induced by paced mating behavior in female rats. PMID- 15113262 TI - Excitotoxic lesions of the gustatory thalamus spare simultaneous contrast effects but eliminate anticipatory negative contrast: evidence against a memory deficit. AB - Using consummately contrast procedures and the same taste stimuli (0.15% saccharin and 1.0 M sucrose), the authors tested the hypothesis that lesions of the gustatory thalamus disrupt gustatory memory in 2 experiments. In Experiment 1, irrespective of the duration of the intersolution interval (0 s, 30 s, 1 min, 2 min, 4 min, 8 min), thalamic lesions had no influence on the expression of simultaneous contrast effects. In Experiment 2, thalamic lesions abolished anticipatory negative contrast at the 0-s intersolution interval. These results provide no support for the experimental hypothesis. Rather, the data seem best interpreted as a lesion-induced disruption of the comparison mechanism responsible for anticipatory negative contrast. By this analysis, different comparison mechanisms underlie simultaneous and anticipatory contrast effects. PMID- 15113263 TI - Hippocampal lesions modulate both associative and nonassociative priming. AB - In associative priming, rats are more likely to orient to a visual stimulus whose memory has not been recently activated (V1) than to one whose memory has been recently activated (V2). However, rats with excitotoxic hippocampal lesions are more likely to orient to the primed V2 than to the unprimed V1. This study investigated the influence of hippocampal lesions on nonassociative priming. Rats received presentations of 2 visual stimuli, V1 and V2, that had been presented more (V2, primed) or less (V1, unprimed) recently. Control rats oriented to V1 rather than to V2, whereas hippocampal rats oriented to V2 rather than to V1. These results parallel those observed in an associative priming procedure and thereby suggest that the role of the hippocampus in priming is general. PMID- 15113264 TI - Inactivation of dorsolateral striatum impairs acquisition of response learning in cue-deficient, but not cue-available, conditions. AB - Rats received bilateral injections of lidocaine or artificial cerebrospinal fluid (aCSF) into the doisolateral striatum 6 min prior to training in either a plus- or T-shaped maze under cue-poor or cue-available conditions. Lidocaine injections significantly impaired acquisition in the cue-poor environments, but not in the cue-available environments. In addition, aCSF control rats trained in a plus-maze in a cue-poor environment reached criterion much more rapidly than did rats trained in a cue-available environment. These findings suggest that cue availability can permit acquisition of response learning in a manner that is not dependent on activity of the striatum. However, in a cue-poor environment, alternate strategies may be less readily available, revealing more efficient striatal involvement in response learning. PMID- 15113265 TI - Electrical stimulation of medial prefrontal cortex reduces conditioned fear in a temporally specific manner. AB - The authors recently showed that extinction of auditory fear conditioning leads to potentiation of tone-evoked activity of neurons in the infralimbic (IL) subregion of the medial prefrontal cortex, suggesting that IL inhibits fear after extinction (M. R. Milad, & G. J. Quirk, 2002). In support of this finding, pairing conditioned tones with brief (300-ms) electrical stimulation of IL reduces conditioned freezing. The present study showed that IL stimulation inhibits freezing if given 0.1 s after tone onset (the latency of tone-evoked responses) but has no effect if given either 1 s before or 1 s after tone onset. This suggests that IL gates the response of downstream structures such as the amygdala to fear stimuli. PMID- 15113266 TI - Temporal encoding in fear conditioning revealed through associative reflex facilitation. AB - Temporal encoding in Pavlovian fear conditioning was examined through conditional facilitation of the short-latency (Rl) component of the rat eyeblink reflex. Rats were fear-conditioned to a tone conditional stimulus (CS) with either a 3- or 9-s interstimulus interval (ISI) between CS onset and the onset of the grid-shock unconditional stimulus (US). Rl facilitation was tested over 2 days, in counterbalanced order, at a latency of 3 s and 9 s from CS onset. CS-produced Rl facilitation, the conditional response (CR), was 3-4 times larger when the test latency equaled the conditioning ISI. These results, coupled with the known neurophysiology of Rl facilitation, suggest that this CR could disclose differences in the time course of CS-generated output from the amygdala when driven by cortical versus subcortical CS-CR pathways. PMID- 15113267 TI - Theta-contingent trial presentation accelerates learning rate and enhances hippocampal plasticity during trace eyeblink conditioning. AB - Hippocampal theta activity has been established as a key predictor of acquisition rate in rabbit (Orcytolagus cuniculus) classical conditioning. The current study used an online brain--computer interface to administer conditioning trials only in the explicit presence or absence of spontaneous theta activity in the hippocampus-dependent task of trace conditioning. The findings indicate that animals given theta-contingent training learned significantly faster than those given nontheta-contingent training. In parallel with the behavioral results, the theta-triggered group, and not the nontheta-triggered group, exhibited profound increases in hippocampal conditioned unit responses early in training. The results not only suggest that theta-contingent training has a dramatic facilitory effect on trace conditioning but also implicate theta activity in enhancing the plasticity of hippocampal neurons. PMID- 15113268 TI - A new method for comparing orthonasal and retronasal olfaction. AB - Odors seem to be perceived differently when presented ortho- or retronasally. In this study, stimuli of controlled concentration and duration were applied in either the anterior nasal cavity or epipharynx by means of air-dilution olfactometry. Stimulus concentration was monitored in the olfactory cleft. In Experiment 1, odor thresholds to a food (chocolate) and a nonfood (lavender) odor were lower for orthonasal, compared with retronasal, stimulation. In Experiment 2, intensity ratings to suprathreshold odor concentrations were significantly higher for orthonasal than for retronasal stimulation with hydro- hydrogen sulfide, but not phenyl ethyl alcohol. Accordingly, amplitudes and latencies of olfactory event-related gen potentials to retronasal stimuli were found to be smaller and prolonged, respectively. This indicates differential processing of olfactory stimuli presented through the retronasal or orthonasal routes. ) PMID- 15113269 TI - Differential effects of adrenergic and corticosteroid hormonal systems on human short- and long-term declarative memory for emotionally arousing material. AB - The effects of adrenergic and corticosteroid hormonal systems on emotional memory were measured in 64 young men. Placebo, propranolol (40 or 80 mg; beta blocker), or metyiapone (corticosteroid synthesis inhibitor) was administered before the viewing of a story composed of emotional and neutral segments. Short- and long term declarative memory for the story was assessed. Propranolol 40 mg had no effects on declarative memory. Propranolol 80 mg impaired short- and long-term declarative memory for emotionally arousing material. Metyrapone did not impair short-term declarative memory but impaired long-term declarative memory for emotionally arousing and neutral material. Results demonstrate that adrenergic and corticosteroid hormonal systems differentially affect declarative memory for emotionally arousing and neutral material, and suggest that interactions between adrenal hormonal systems modulate emotionally arousing declarative memory in humans. PMID- 15113270 TI - Anteromedial temporal lobe damage blocks startle modulation by fear and disgust. AB - The acoustic startle reflex (ASR) is potentiated during negative emotion, but attenuated during positive emotional experience. The modulation of the ASR by fear depends critically on the amygdala. The authors investigated ASR modulation to fearful, disgusting, pleasant, and neutral stimuli in 12 patients with unilateral damage to the anteromedial temporal lobe including the amygdala (6 left, 6 right), 1 patient with bilateral temporal lobe damage including the amygdala, and 12 comparison participants. Both groups with unilateral damage, as well as the subject with bilateral damage, showed a complete lack of ASR potentiation to both fear and disgust stimuli. The findings suggest that potentiation of the ASR by disgust and fear depends on the integrity of the anteromedial temporal lobe. PMID- 15113271 TI - An implicit learning task activates medial temporal lobe in patients with Parkinson's disease. AB - Recent studies provide evidence for an interaction between a declarative memory system, dependent on the medial temporal lobe (MTL), and a habit memory system, dependent on the basal ganglia. Using functional MRI, the authors studied this interaction when 1 system was compromised by neurological disease. Neural activity when performing a habit-learning task was compared in normal controls and subjects with Parkinson's disease (PD). Patients with PD showed less activation in the caudate nucleus and greater activation in a region of prefrontal cortex that has been associated with explicit memory retrieval. Patients with PD also showed activation of the MTL during the weather-prediction task. These findings are consistent with an interaction between memory systems of the MTL and the striatum. PMID- 15113272 TI - Chemical lesion of the bed nucleus of the stria terminalis blocks the behavioral consequences of uncontrollable stress. AB - Uncontrollable or inescapable shock (IS) produces behavioral changes that are characterized by a sensitized fear system and a deficit in fight-flight responding. These behavioral changes have been argued to represent an anxiety like state produced by the uncontrollability of the stressor. The bed nucleus of the stria terminalis (BNST) has been implicated in the mediation of long-duration responses to unpredictable stressors, which have also been argued to represent anxiety. In the present study, the effects of BNST chemical lesion on the IS induced sensitization of freezing to an environment previously paired with shock and the IS-induced impairment of escape responding were investigated. BNST chemical lesion blocked the potentiation of freezing and the increases in escape latency that normally follow IS. PMID- 15113274 TI - Factors related to activity limitations in a group of Cuban Americans before and after hip fracture. AB - BACKGROUND AND PURPOSE: Cuban Americans over the age of 50 years, like people from a number of other cultures, have been exposed to distinctive social and psychological aspects of their culture that may affect their recovery from a major health event. The purpose of this study was to determine factors related to activity limitations in a group of Cuban Americans recovering from hip fractures. SUBJECTS: The subjects were 28 adults of Cuban-American origin with an average age of 79 years (SD=9, range=57-96) who were recovering from hip fracture. Subjects were followed for 2 months after discharge from a hospital-based inpatient rehabilitation facility. METHODS: Subjects were interviewed at the time of discharge from the rehabilitation facility and 2 months after discharge, and they completed the Medical Outcomes Study 36-Item Short-Form Health Survey (SF 36) and a social support interview. Functional Independence Measure scores and information regarding rehabilitative care were obtained from the rehabilitation facility. Multilinear regression analysis was used to determine the factors related to activity limitations before and after hip fracture. RESULTS: More premorbid activity limitations were related to poorer mental health status, older age, and male sex. More activity limitations at the time of discharge from the rehabilitation hospital were related to poorer mental health status prior to admission and partial or non-weight-bearing status after hip fracture. Older age and male sex were premorbid factors that appeared to relate to function after discharge. More activity limitations during the 2 months after discharge were related to the subjects reporting more social support in the form of physical assistance, less social support in the form of emotional assistance, less negative social support in the form of demand and criticism, status for weight bearing as tolerated at the time of discharge from the rehabilitation hospital, and older age. DISCUSSION AND CONCLUSION: In this group of Cuban Americans, mental health status was related to more activity limitations prior to admission to the rehabilitation facility and early in the recovery process, but not later in the recovery process. Conversely, social support was related to activity limitations later in the recovery process, but not prior to admission or early in the recovery process. Physical therapists should be aware of the psychosocial determinants of activity limitations during the process of recovery from disease or trauma. PMID- 15113275 TI - Gross motor capability and performance of mobility in children with cerebral palsy: a comparison across home, school, and outdoors/community settings. AB - BACKGROUND AND PURPOSE: Mobility of children with cerebral palsy (CP) has generally been examined in terms of capability (what a child can do) in a controlled environment, rather than performance (what a child does do) in everyday settings. The purpose of this study was to compare gross motor capability and performance across environmental settings in children with CP. SUBJECTS: The subjects were 307 children with CP, aged 6 to 12 years, who were randomly selected across Ontario, Canada. METHODS: Children were grouped by capability (the highest of 3 items achieved on the Gross Motor Function Measure). Performance was measured via a parent-completed questionnaire on usual mobility methods in the home, at school, and in the outdoors or community. RESULTS: There were statistically significant differences in performance across settings for children in all capability groups. Children who were capable of crawling performed crawling more at home than at school or in the outdoors or community. Children who were capable of walking with support performed walking with support more at school than in the outdoors or community. Children who were capable of walking alone performed walking alone more at home than at school or in the outdoors or community, and more at school than in the outdoors or community. DISCUSSION AND CONCLUSION: The results provide evidence that children with CP with similar capability demonstrate differences in performance across settings. The results suggest that physical therapists should examine performance in the settings that are important to the child's daily life. PMID- 15113276 TI - Psychometric properties of 2 simplified 3-level balance scales used for patients with stroke. AB - BACKGROUND AND PURPOSE: To determine if the 3-level scales yield data as reliable and valid as data obtained for the original scales, 2 simplified 3-level measures of balance--a modified Berg Balance Scale (BBS-3P) and a modified Postural Assessment Scale for Stroke Patients (PASS-3P)--were proposed by the researchers, and psychometric properties of each were compared with those of the original measures (the Berg Balance Scale [BBS] and the Postural Assessment Scale for Stroke Patients [PASS], respectively) in patients with stroke. SUBJECTS AND METHODS: The study consisted of 2 parts. The first part examined the reliability and concurrent and convergent validity of measurements obtained with these instruments. A total of 77 patients participated in this part of the study. The 3 levels in the center of the BBS were collapsed to a single level (ie, 0-2-4) to form the BBS-3P. Similarly, the 2 middle scores of the center of the PASS were averaged (ie, 0-1.5-3) to form the PASS-3P. In the second part of the study, the predictive validity and responsiveness of these measures were examined. The BBS and PASS scores of 226 patients were retrieved from the records of participants in the Quality of Life After Stroke Study, and these scores were converted into the proposed BBS-3P and PASS-3P scores. RESULTS: The BBS-3P and PASS-3P showed high concurrent validity with the BBS and PASS, good predictive validity for disability, and moderate to high responsiveness. Importantly, the psychometric properties of the BBS-3P and PASS-3P were essentially identical to those of the original BBS and PASS. DISCUSSION AND CONCLUSION: The psychometric properties of both simplified 3-level balance measures were comparable to those of the full, nontruncated scales. Future study is needed to investigate how much meaningful utility can be gained from the scheme of simplification of scaling. PMID- 15113277 TI - Childrearing practices and developmental expectations for Mexican-American mothers and the developmental status of their infants. AB - BACKGROUND AND PURPOSE: The impact of parent education programs on early intervention programs is not thought to be uniform among children from majority and minority populations. This study examined the relationship between maternal childrearing practices and behaviors and the developmental status of Mexican American infants. SUBJECTS: Participants were 62 Mexican-American mother-infant pairs. The infants' mean adjusted age was 12 months (SD=1.7, range=9-14). A third of the children were diagnosed with developmental delays and referred for early intervention by physicians or therapists when the children received their medical follow-up. The group was stratified according to socioeconomic status and acculturation using the Bidimensional Acculturation Scale for Hispanics. This scale uses cutoff points to classify individuals into 3 levels of acculturation. METHODS: Information on childrearing practices and behaviors was gathered using the Parent Behavior Checklist (PBC), the Home Observation for Measurement of the Environment (HOME) Inventory, and the Nursing Child Assessment Teaching Scale (NCATS). Infants' developmental status was assessed by use of the Bayley Scales of Infant Development II (BSID II). The Pearson product moment correlation, partial correlations, Fisher z transformation, and multiple regression analyses were used to examine the relationship between childrearing practices and parenting behaviors, demographic factors, and infants' developmental status. RESULTS: Maternal nurturing behaviors, parent-child interaction, and quality of the home environment were positively correlated with the infants' cognitive development. Maternal years of education modified the observed relationship between PBC and BSID II scores but not the observed relationship between HOME Inventory and NCATS scores. The childrearing practices, maternal socioeconomic status (SES) and age, and infants' gestational age at birth (GA) explained 45% of the variance in infants' cognitive scores. The infants' GA, maternal SES and age, and NCATS scores accounted for 32% of the motor scores on the BSID II. DISCUSSION AND CONCLUSION: The findings partially support a link between aspects of the mothers' childrearing behaviors and their infants' cognitive developmental status. For motor developmental status, the association appeared stronger with the infants' characteristics than with maternal childrearing practices and behaviors tested in this study. PMID- 15113278 TI - End-stage renal disease in people with type 2 diabetes: systemic manifestations and exercise implications. PMID- 15113279 TI - Does evidence exist on whether specific interventions can improve adherence to a home exercise program in a patient with intermittent claudication? PMID- 15113280 TI - Sensitivity of bone to glucocorticoids. AB - Glucocorticoids are used widely in a range of medical specialities, but their main limitation is an adverse impact on bone. Although physicians are increasingly aware of these deleterious effects, the marked variation in susceptibility between individuals makes it difficult to predict who will develop skeletal complications with these drugs. Although the mechanisms underlying the adverse effects on bone remain unclear, the most important effect appears to be a rapid and substantial decrease in bone formation. This review will examine recent studies that quantify the risk of fracture with glucocorticoids, the mechanisms that underlie this increase in risk and the potential basis for differences in individual sensitivity. An important determinant of glucocorticoid sensitivity appears to be the presence of glucocorticoid-metabolizing enzymes within osteoblasts and this may enable improved estimates of risk and generate new approaches to the development of bone-sparing anti-inflammatory drugs. PMID- 15113281 TI - Binding of alpha2-macroglobulin to GRAB (Protein G-related alpha2-macroglobulin binding protein), an important virulence factor of group A streptococci, is mediated by two charged motifs in the DeltaA region. AB - GRAB (Protein G-related alpha2M-binding protein) is a surface protein of group A streptococci and exhibits high affinity for alpha2-macroglobulin (alpha2M), a broad-range protease inhibitor. It is the sole alpha2M-binding protein of group A streptococci that has been shown to promote bacterial virulence in a mouse model of skin infection. The binding site for alpha2M was predicted to be in the N terminal A domain of GRAB. In the present study, the alpha2M-binding domain was first narrowed down to 34 amino acids (amino acids 34-67) using variable truncated N-terminal GRAB fusion proteins. The sequence of the identified domain was used to design overlapping synthetic peptides of different sizes, which were then immobilized on a membrane and assayed for their alpha2M-binding activity. The peptide screening revealed two binding motifs of ten amino acids length, located in the DeltaA (N-terminal part of the A domain) region (amino acids 34 67) with the sequences PRIIPNGGTL (amino acids 41-50) and NAPEKLALRN (amino acids 56-65) respectively. These motifs were used for systematic mutational analysis by generating synthetic peptides containing individual amino acid substitutions at every position of the mapped binding regions. The results indicated a critical role for the arginine residue at position 42 in the first binding domain and at position 64 in the second binding region. Validation of arginine residues as the critical amino acids for alpha2M binding was achieved by site-directed mutagenesis and binding assays. Competitive inhibition assays with GRAB containing amino acid substitutions R42G (Arg42-->Gly), R64G and R42G/R64G indicated differential contribution of the arginine residues at positions 42 and 64 to alpha2M-binding activity and, thus, their involvement in GRAB-induced virulence. PMID- 15113282 TI - Management of dyschromias in ethnic skin. AB - Pigmentary disorders are one of the most common skin disorders among people of color. Dyspigmentation in the form of either hyperpigmentation or hypopigmentation is often psychologically devastating to patients with darker skin. There is marked contrast between normally pigmented hyperpigmented, hypopigmented or depigmented skin in people of color. Despite being common, pigmentary disorders remain difficult to treat. PMID- 15113283 TI - The medical and surgical therapy of pseudofolliculitis barbae. AB - Pseudofolliculitis barbae (PFB) is a common, chronic, inflammatory skin disorder seen mainly in individuals with curly hair. This condition is seen most frequently in black men who shave their beards but may also be seen in women of all races who wax or shave the axillary and pubic skin. The etiology of PFB is multifactorial, and heretofore a cure has been considered impossible for those desiring a clean-shaven face. The following article serves to discuss the current medical and surgical therapies available for this condition. Medical treatments for this condition include various combinations of topical antibiotics, corticosteroids, and retinoids. In the surgical arena, laser therapy has revolutionized the treatment of PFB and has enabled cure for the first time for those plagued by this disorder and for whom a beardless face is acceptable. PMID- 15113284 TI - Medical and surgical therapies for alopecias in black women. AB - Hair loss is a common problem that challenges the patient and clinician with a host of cosmetic, psychological and medical issues. Alopecia occurs in both men and women, and in all racial and ethnic populations, but the etiology varies considerably from group to group. In black women, many forms of alopecia are associated with hair-care practices (e.g., traction alopecia, trichorrhexis nodosa, and central centrifugal cicatricial alopecia). The use of thermal or chemical hair straightening, and hair braiding or weaving are examples of styling techniques that place African American women at high risk for various "traumatic" alopecias. Although the exact cause of these alopecias is unknown, a multifactorial etiology including both genetic and environmental factors is suspected. A careful history and physical examination, together with an acute sensitivity to the patient's perceptions (e.g., self-esteem and social problems), are critical in determining the best therapy course. Therapeutic options for these patients range from alteration of current hair grooming practices or products, to use of specific medical treatments, to hair replacement surgery. Since early intervention is often a key to preventing irreversible alopecia, the purpose of the present article is to educate the dermatologist on all aspects of therapy for hair loss in black women--including not only a discussion of the main medical and surgical therapies but also an overview of ethnic hair cosmetics, specific suggestions for alterations of hair-care practices, and recommendations for patient education and compliance. PMID- 15113285 TI - Laser-assisted hair removal for darker skin types. AB - Before the advent of longer wavelengths, longer pulse durations and more efficient cooling devices, laser-assisted hair removal was best suited for phototypes I-III with dark terminal hairs. Now, laser-assisted hair removal can be performed safely and efficaciously on darker skin types. The long-pulsed diode and Nd:YAG-wavelength-based laser systems are best suited to safely and effectively treat patients with darker skin types. By using conservative fluences, longer pulse durations and multiple treatments, safe and effective laser-assisted hair removal is possible for darker skin types. Patients with darker skin who suffer from hirsutism, hypertrichosis, and pseudofolliculitis barbae can finally be treated safely and effectively. PMID- 15113287 TI - Chemical peeling in ethnic/dark skin. AB - Chemical peeling for skin of color arose in ancient Egypt, Mesopotamia, and other ancient cultures in and around Africa. Our current fund of medical knowledge regarding chemical peeling is a result of centuries of experience and research. The list of agents for chemical peeling is extensive. In ethnic skin, our efforts are focused on superficial and medium-depth peeling agents and techniques. Indications for chemical peeling in darker skin include acne vulgaris, postinflammatory hyperpigmentation, melasma, scarring, photodamage, and pseudofolliculitis barbae. Careful selection of patients for chemical peeling should involve not only identification of Fitzpatrick skin type, but also determining ethnicity. Different ethnicities may respond unpredictably to chemical peeling regardless of skin phenotype. Familiarity with the properties each peeling agent used is critical. New techniques discussed for chemical peeling include spot peeling for postinflammatory hyperpigmentation and combination peels for acne and photodamage. Single- or combination-agent chemical peels are shown to be efficacious and safe. In conclusion, chemical peeling is a treatment of choice for numerous pigmentary and scarring disorders arising in dark skin tones. Familiarity with new peeling agents and techniques will lead to successful outcomes. PMID- 15113286 TI - Acne in ethnic skin: special considerations for therapy. AB - Acne vulgaris occurs in people of all ethnicities and races. Although the pathophysiology and treatment options are similar in all skin phototypes, darker skinned patients have higher incidence rates of two sequelae of acne: postinflammatory hyperpigmentation and keloidal scarring. Postinflammatory hyperpigmentation may also be triggered by skin irritation. In choosing therapies for patients of color, therefore, clinicians must find a balance between aggressive early intervention to target inflammatory acne lesions, and gentle treatments to increase tolerability and avoid skin irritation. For most patients, a combination of topical retinoids, and topical or oral antibiotics with hydroquinone (as needed) to control hyperpigmentation will be successful. For patients with sensitive skin, topical agents in lower concentrations and cream vehicles are preferred. If tolerated, the retinoid strength can be titrated upward after four to six weeks. Ethnic patients also need to be counseled on use of noncomedogenic and nonirritating skin and hair-care products. Individualized care and close monitoring is required. PMID- 15113288 TI - The aging face in patients of color: minimally invasive surgical facial rejuvenation-a targeted approach. AB - We are in the midst of exciting times from a demographic standpoint as the population of darker-skinned people grows exponentially in the United States. Although there is a growing demand for facial cosmetic procedures amongst people of color, the total number of individuals undergoing surgical facial rejuvenation is dwarfed by the current Caucasian market. In order to provide optimal options for facial rejuvenation, cosmetic surgeons must have an underlying appreciation for the dynamic interplay among ethnicity, facial morphology, and the progression of aging. The purpose of the present article is to outline the minimally invasive surgical options for facial rejuvenation best suited for the patient of color. Although the population of individuals with darker-pigmented skin is quite vast, the present paper will focus on individuals of African descent, with whom the author has the most experience. Preferred surgical techniques include blepharoplasty, autologous facial fat transplantation, percutaneous cheek lift, and submental liposuction. Cutaneous surgeons familiar with soft-tissue surgery and facial anatomy should feel comfortable performing these techniques. PMID- 15113289 TI - Medical and surgical therapies for keloids. AB - Keloids are benign, but sometimes painful and/or pruritic, proliferative growths of dermal collagen, usually resulting from excessive tissue response to trauma. Although benign, the social and psychological impact on affected individuals must be considered. Keloids often arise secondary to ear piercing and operative procedures. No single treatment modality is always successful. The more common ones are discussed. Some of the medical therapies include corticosteroids, interferon, 5-fluorouracil, and imiquimod. Primary excision and cryosurgery are among the major surgical options. Radiation therapies and other physical modalities are also discussed. PMID- 15113291 TI - Radioresistant laryngeal cancer: beyond the TNM stage. AB - Early stage squamous cell carcinoma of the larynx can be effectively cured by radiotherapy. Unfortunately treatment failures do occur and at present cannot be predicted by the clinician. This article reviews the potential molecular and cellular markers that may help to predict radioresistance in early stage laryngeal cancer. PMID- 15113292 TI - Long-term hearing results of incus transposition. AB - Incus transposition is an option for reconstructing an ossicular discontinuity. We looked at the long-term outcome (average 9.2 years) in terms of hearing results in patients who had undergone incus transposition. Patients were requested to attend for a pure tone audiogram. Details of their preoperative audiograms were obtained from the patients' case notes. A postoperative air-bone gap of 20 dB or less averaged over four frequencies (0.5, 1, 2, 4 kHz) was considered to be an acceptable result. We found that 70% of patients had an air bone gap of 20 dB or less and 54% had had cholesteatoma surgery at the time of the incus transposition. This study represents the longest follow-up of patients having undergone incus transposition from our search of the literature. Incus transposition, we feel, offers good long-term hearing results. PMID- 15113293 TI - What is the impact of consultant supervision on outpatient follow-up rate? AB - This study investigated the impact of consultants on recycling rates of patients in the ENT outpatient clinic. A retrospective case review of 4205 consecutive patients who attended ENT outpatient clinics of an UK teaching hospital over a 3 month period was conducted. There was a significant association between grade of medical staff and recycling rate of new patients, and also for review patients. Junior doctors have lower recycling rates in consultant-led clinics compared with clinics in the absence of consultants for both new patients (consultant-led 41.0%, without consultant 60.1%; P < 0.01) and old patients (consultant-led 48.9%, without consultant 65.0%; P < 0.01). Individual consultant's practice was reflected upon the overall recycling rate of the clinic as a whole (r = 0.94, P = 0.001). In conclusion, individual consultant's practice dictated recycling rate in the ENT outpatient clinic. Junior doctors were less likely to make follow-up appointments when directly supervised by their consultants. PMID- 15113294 TI - Interobserver perceptual analysis of smokers voice. AB - The purpose of this study was to investigate variations in the voice by three experienced speech-language therapists. Forty-eight men and nine women from the Associacao dos Alcoolicos Anonimos, Santos were studied. Their ages were from 28 to 81 years, with median of 49 years and everyone was a smoker for 60 to 720 months. Most of them used more than 20 cigarettes a day and all of them had stopped alcohol use for 1 to 25 months. The perceptual analysis of the voices was performed by means of the GRBAS scale. The voice sample consisted of a sustained vowel /a/ at a comfortable pitch and loudness level. The three judges were blinded to the opinion of their colleagues and a kappa test was applied. For roughness, the concordance rates were 59.6% for observers 1 and 2 (kappa = 0.234); 47.4% for 1 and 3 (kappa = 0.047) and 52.6% (kappa = 0.016) for 2 and 3. For breathiness, the concordance rates were 80.7% for observers 1 and 2 (kappa = 0.191); 57.9% for 1 and 3 (kappa = 0.147) and 57.9% (kappa = 0.156) for 2 and 3. With regard to asthenic quality, there was concordance of 100% for observers 1 and 2, so kappa could not be applied; the concordance rate was 96.5% for 1 and 3 and for 2 and 3. The evaluation for strained voice revealed concordance rates of 71.9% for observers 1 and 2 (kappa = -0.017); 59.6% for 1 and 3 (kappa = 0.095) and 70.2% (kappa = 0.039) for 2 and 3. The disagreement among the observers was worst for pathological rather than normal voices; when disagreement was present among experienced judges, it was of only one point in the scale used. PMID- 15113295 TI - An objective evaluation of the waterproofing qualities, ease of insertion and comfort of commonly available earplugs. AB - Earplugs are commonly recommended to protect the middle ear from water in patients with exposed middle ears. This study assessed the waterproofing qualities, ease of insertion and comfort of six commonly available earplugs. Ten subjects (20 ears) were assessed by placing a pre-weighed neurosurgical pattie in their cleaned ear canal under microscope guidance. The subjects underwent a standardized head wetting regime. The outer ear was dried, earplug and pattie removed and pattie reweighed. The difference in weight was calculated. The subjects were also asked to score the difficulty of insertion and comfort of the earplugs on a visual analogue scale. The results show a significant difference in the waterproofing qualities of the various types of earplugs. Cotton wool with petroleum jelly was the most effective (P < 0.001). It was also the easiest to insert and the most comfortable for the subject (P < 0.001). PMID- 15113296 TI - Vibration characteristics and function of atelectatic segments in the tympanic membrane in fresh human cadaveric temporal bones. AB - Dimeric segments are commonly encountered in otological practice. They may be associated with a mild conductive hearing loss and often coexist with other tympanic membrane and middle ear abnormalities. Some otologists have advocated surgical management but the consequences of dimeric segment stiffening, shielding or excision and grafting on acoustic transfer to the stapes footplate has been poorly explored. In this study, laser Doppler vibrometry was used to measure vibrations at the tympanic membrane and the stapes footplate in the fresh cadaveric human temporal bone. The dimeric segment vibrates more than the adjacent, thicker normal tympanic membrane. Shielding or excision and grafting of the dimeric segment with thicker and stiffer materials has little effect on displacement at the stapes footplate. PMID- 15113297 TI - The effect of the steroid sex hormones on the nasal airway during the normal menstrual cycle. AB - The changes in the nasal mucosa during the menstrual cycle have been little researched and the role of oestrogens debated. The aim of this study was to measure a spectrum of physiological parameters in relation to the peak and trough of these hormonal levels. Ten women underwent measurements at the onset of menses and at the time of ovulation. On each occasion, anterior rhinoscopy, peak inspiratory nasal flow, acoustic rhinometry, anterior rhinomanometry, mucociliary clearance time and rhinitis questionnaire score were recorded. All measurements except anterior rhinoscopy showed a difference between the two readings consistent with nasal congestion at the peri-ovulatory stage of the cycle, of which anterior rhinomanometry and mucociliary time were significant (P 5%. No patients experienced phototoxicity, hepatic or renal dysfunction, tendonitis or crystalluria. Oral gatifloxacin 400 mg once-daily appeared to be a safe and effective alternative to clarithromycin in the treatment of community-acquired pneumonia. PMID- 15113317 TI - The plasma level of soluble urokinase receptor is elevated in patients with Streptococcus pneumoniae bacteraemia and predicts mortality. AB - This multicentre prospective study was conducted to investigate whether the level of the soluble form of urokinase-type plasminogen activator receptor (suPAR) is elevated during pneumococcal bacteraemia and is of predictive value in the early stage of the disease. Plasma levels of suPAR were increased significantly (median 5.5; range 2.4-21.0 ng/mL) in 141 patients with pneumococcal bacteraemia, compared to 31 healthy controls (median 2.6, range 1.5-4.0 ng/mL, p 0.001). Furthermore, suPAR levels were elevated significantly in patients who died from the infection (n = 24) compared to survivors (n = 117; p < 0.001). No correlation was found between suPAR levels and C-reactive protein. In univariate logistic regression analysis, hypotension, renal failure, cerebral symptoms and high serum concentrations of protein YKL-40 and suPAR were associated significantly with mortality (p < 0.05). In multivariate analysis, only suPAR remained a significant predictor of death (mortality rate of 13 for suPAR levels of > 10 ng/mL; 95% CI: 1.1-158). The increase in suPAR levels may reflect increased expression by vascular or inflammatory cells in the setting of pneumococcal sepsis. This plasma protein may be used to identify patients who are severely ill with pneumococcal bacteraemia. PMID- 15113318 TI - Evaluation of the OSIRIS video reader as an automated measurement system for the agar disk diffusion technique. AB - Measurement of inhibition zones by the automated OSIRIS system was compared with manual measurement. In total, 14 176 measurements were made with 352 staphylococcal and 80 Enterobacteriaceae isolates, involving four panels of antibiotics on round and square Mueller-Hinton agar plates, according to the German DIN 58940 recommendations. Variations of +/- 3 mm in zone size measurements were defined as tolerable. Very major errors (i.e., classification of a resistant isolate as susceptible by the OSIRIS system) occurred in < 1% of tests. With staphylococci, the best concordance was recorded for rifampicin (91.3%), moxifloxacin (88.1%), and gentamicin (86.3%), while the concordance on square plates for vancomycin, pristinamycin and kanamycin was 97.2%, 96.1% and 96.0%, respectively. The poorest concordance was for cefuroxime (43.7%) and novobiocin (47.0%) on round plates, and fosfomycin (36.5%) and chloramphenicol (84.0%) on square plates. With Enterobacteriaceae, 100% concordance was recorded for ampicillin, gentamicin and ciprofloxacin on round agar plates, and for gentamicin, cefoxitin and nalidixic acid on square plates. The poorest results were recorded for nalidixic acid (32.5%) and piperacillin (82.5%) on round plates, and for nitrofurantoin (72.5%) and amoxycillin (82.5%) on square plates. It was concluded that the OSIRIS system was a rapid and reliable system for measuring disk susceptibility test results on round and square agar plates. PMID- 15113319 TI - Evaluation of Bacillus anthracis extractable antigen for testing anthrax immunity. AB - Three extractable Bacillus anthracis cell-wall-associated antigens were evaluated for potential use as skin testing agents, and as possible candidates for in-vitro diagnosis of anthrax immunity. Anthraxin and a partially purified extractable antigen (EAP) were produced from avirulent B. anthracis strain 34F2 (Sterne). The thermoextractable antigen used for the Ascoli reaction was obtained commercially. Guinea-pigs were immunised and boosted several times subcutaneously with the Sterne live veterinary anthrax vaccine. Four weeks after the last booster dose, animals were skin-tested with the three antigens. Serum antibody levels were also determined by ELISA, and the in-vitro T-cell response was evaluated by [3H] thymidine incorporation. EAP was the most active antigen in both the serological and cellular reactions. EAP also elicited a distinct positive skin reaction in animals immunised with B. anthracis. The data obtained in this preliminary study indicated that extractable cell-wall antigens obtained from the vegetative form of B. anthracis may be used for skin tests and in-vitro testing of specific humoral and cell-mediated anthrax immunity. PMID- 15113320 TI - Empirical data on the varicella situation in Germany for vaccination decisions. AB - As epidemiological data concerning varicella in Germany were not available, a retrospective study was conducted to investigate the impact of the disease, focusing also on economic aspects. A representative German-wide sample of 1334 unvaccinated varicella cases was obtained in 1999 from randomly selected paediatric (P) and general, as well as internal, mainly adult (A), practices. Following representative weighting, the median age was 5 years, with 90% of cases aged < 12 years. The highest incidence was in children aged 5-6 years. Varicella related complications occurred in 5.7% of patients, and accounted for 0.1 hospital days/case on average. Certificates of sick leave were issued for 1.3 sick days/case, with 0.6 days paid by health insurance funds to parents caring for their sick child, and 0.7 days paid by the employer. With an annual incidence of 760,000 diagnosed cases in Germany for the year 1999, this amounts to an annual cost of c.150 million Euro, with c. 50 million Euro paid by the statutory health insurance system. It was concluded that universal varicella vaccination in Germany would provide essential clinical improvements for patients and prevent hospital admissions. In addition, significant economic benefits can be expected, mainly because of the high level of indirect health costs in Germany. PMID- 15113321 TI - Intravenous catheter infections associated with bacteraemia: a 2-year study in a university hospital. AB - The aim of this retrospective study was to assess the incidence and aetiology of central and peripheral venous catheter (C/PVC) infections during a 2-year period (1999-2000) and to determine the susceptibility of isolated microorganisms to various antimicrobial agents. Catheter tips were processed using the semiquantitative method and blood cultures were performed with the BacT/Alert automated system. Antibiotic susceptibilities were performed by disk agar diffusion and MICs were determined by Etest, according to NCCLS standards. During the study period, samples from 1039 C/PVC infections were evaluated, yielding 384 (37.0%) positive cultures. Blood cultures were also available from 274 patients, of which 155 (56.6%) yielded the same microorganism as from the catheter. No bloodstream infections were detected in 104 C/PVC-positive cases. Methicillin resistant coagulase-negative staphylococci were the most frequent isolates, followed by Gram-negative bacteria, especially Pseudomonas aeruginosa. Resistance to glycopeptides among staphylococci and enterococci was not detected, whereas 60% of Gram-negative bacilli were resistant to beta-lactams. PMID- 15113322 TI - Prevalence of colonisation with third-generation cephalosporin-resistant Enterobacteriaceae in ICU patients of Heidelberg University Hospitals. AB - The aim of this study was to assess colonisation and transmission of third generation cephalosporin-resistant Enterobacteriaceae (CRE) from patients in 16 intensive care units. A prospective, repetitive point prevalence survey was performed over 6 months, involving samples from 1851 patients. CRE were isolated from 186 (10%) patients, with Enterobacter spp. being the most common. Mean point prevalence rates were significantly higher for paediatric wards (22.5%) compared to surgical (8.1%) and medical (5.5%) units. All CRE isolates were typed by pulsed-field gel electrophoresis. Non-outbreak nosocomial transmission rates of these pathogens were calculated as 12.8% for paediatric patients, compared to 6.8% for adult patients, which may reflect differences in sensitivity to overgrowth with resistant bacteria and contact with health care workers. PMID- 15113323 TI - Microbiological and epidemiological studies of Enterococcus faecium resistant to amoxycillin in a university hospital in eastern France. AB - Over 3 years, during six 1-month periods of surveillance, 69 cases of Enterococcus faecium colonisation or infection were detected in a university hospital in eastern France. Thirty-two cases involved strains resistant to amoxycillin (crude incidence of 0.21/1000 patient-days). The risk of infection with E. faecium was higher if the patient was hospitalised in a haematology ward and/or treated with cephalosporins. Amoxycillin-resistant isolates (AmRE) were isolated from different wards and time periods, and none of the characteristics studied were associated significantly with amoxycillin resistance. Amoxycillin sensitive and -resistant isolates were characterised by pulsed-field gel electrophoresis. Three epidemic patterns were identified which contained 87.5% (28/32) of the AmRE isolates, indicating that clonal spread was responsible, at least partially, for the high incidence of AmRE in this hospital. PMID- 15113324 TI - Methicillin-resistant Staphylococcus aureus (MRSA) with reduced susceptibility to glycopeptides (GISA) in 63 French general hospitals. AB - Clinical isolates of methicillin-resistant Staphylococcus aureus (n = 1070) collected from 63 French general hospitals during June 2000 (n = 1070) were screened initially for reduced susceptibility to glycopeptides (GISA) on brain heart infusion agar containing teicoplanin 6 mg/L. Glycopeptide MICs were determined for the 145 isolates that grew on the screening plates. Of the 1070 isolates, 1.4% were GISA on Mueller-Hinton agar, and 2.9% by Etest with a high inoculum, while 0.7% and 2.9% were GISA by vancomycin and teicoplanin population analysis profiles, respectively. Most isolates were resistant to gentamicin and rifampicin or fosfomycin or fusidic acid, as determined by disk diffusion. Pulsed field gel electrophoresis of the 31 GISA isolates identified four clones, with dissemination of one predominant clone. In these French hospitals there was a low incidence of GISA and hetero-GISA. PMID- 15113325 TI - Effect of comprehensive validation of the template isolation procedure on the reliability of bacteraemia detection by a 16S rRNA gene PCR. AB - The influence of the DNA extraction method on the sensitivity and specificity of bacteraemia detection by a 16S rRNA gene PCR assay was investigated. The detection limit of the assay was 5 fg with purified DNA from Escherichia coli or Staphylococcus aureus, corresponding to one bacterial cell. However, with spiked blood samples, the detection limits were 10(4) and 10(6) CFU/mL, respectively. The sensitivity of the S. aureus assay was improved to the level of the E. coli test with the addition of proteinase K to the commercial DNA extraction kit protocol. Ten (16.6%) of 60 amplification reactions were positive with templates isolated from sterile blood, while PCR reagent controls were negative, thereby indicating contamination during the DNA extraction process. Blood samples were spiked with serial dilutions of E. coli and S. aureus cells, and six PCR results were obtained from three extractions for each blood sample. A classification threshold system was devised, based on the number of positive reactions for each sample. Samples were deemed positive if at least four positive reactions were recorded, making it possible to avoid false-positive results caused by contamination. These results indicate that a comprehensive validation procedure covering all aspects of the assay, including DNA extraction, can improve considerably the validity of PCR assays for bacteraemia, and is a prerequisite for the meaningful detection of bacteraemia by PCR in the clinical setting. PMID- 15113326 TI - Detection of an outbreak of methicillin-resistant Staphylococcus aureus with reduced susceptibility to glycopeptides in a French hospital. AB - Staphylococcus aureus isolates were screened for reduced susceptibility to glycopeptides with an initial glycopeptide agar screening test, followed by confirmation of the strains thus identified by two Etest strip techniques and population analysis. This procedure detected 48 methicillin-resistant S. aureus (MRSA) isolates with reduced susceptibility to glycopeptides from 24 patients among 883 MRSA isolates tested. The dissemination of a single clone was confirmed by pulsed-field gel electrophoresis. PMID- 15113327 TI - Comparison of PCR detection of mecA with agar dilution and Etest for oxacillin susceptibility testing in clinical isolates of coagulase-negative staphylococci. AB - Oxacillin-resistant staphylococci are heterogeneous in their expression of resistance to beta-lactam antibiotics. Different recommendations regarding screening methods for routine use have been published. In this study, the susceptibility to oxacillin of 232 coagulase-negative staphylococci (CoNS) was determined by agar dilution, Etest and presence of the mecA gene. When an oxacillin resistance breakpoint of > or = 0.5 mg/L was used, the sensitivity and specificity for agar dilution were 97.6% and 100%, and those for Etest were 100% and 95.4%. The current National Committee for Clinical Laboratory Standards oxacillin breakpoint recommendation will categorise accurately the CoNS species encountered commonly. PMID- 15113328 TI - Management of long-term catheter-related Brevibacterium bacteraemia. AB - Brevibacterium has been reported as a rare cause of implanted-device infection. In two cases of recurrent Brevibacterium casei bacteraemia associated with infection of surgically implanted intravascular devices, relapse occurred 2 and 5 months, respectively, after completion of therapy with vancomycin via the infected catheter. A second intravenous antibiotic therapy course by the antibiotic-lock technique led to bacteriological cure in one patient. Molecular typing results demonstrated that the recurrent bacteraemia was caused by the same strain. Implanted-device removal may be necessary, in addition to appropriate antibiotics, for successful management of such infections. PMID- 15113329 TI - Cytomegalovirus mononucleosis as a cause of prolonged fever and prominent weight loss in immunocompetent adults. AB - Four immunocompetent adults presented with protracted fever lasting > 6 weeks and severe weight loss, associated with primary cytomegalovirus (CMV) infection. Each patient had spleen enlargement, lymphocytosis and hypertriglyceridaemia, but recovered spontaneously. A further 20 immunocompetent patients with primary CMV infection were also reviewed, and all presented the usual clinical picture of CMV mononucleosis. It was concluded that CMV mononucleosis should be considered in the differential diagnosis in patients with prolonged fever and weight loss if lymphocytosis is present. PMID- 15113330 TI - Molecular and diagnostic clinical virology in real time. PMID- 15113331 TI - Report from the European Conference on the Role of Research in Combating Antibiotic Resistance, 2003. AB - Europe has been at the forefront of efforts to control antibiotic resistance, and this globally important health care problem has prompted numerous recommendations for action at both the national and international levels. Starting in 2002, research on antimicrobial resistance has been considered to be one of the specific objectives of the Sixth Framework Programme (FP6) within the European Union. This report summarises the plenary presentations, as well as the findings of six Working Groups covering specific areas of antibiotic resistance, given at a conference in November 2003 entitled 'The Role of Research in Combating Antibiotic Resistance', co-organised by the European Union and the European Society for Clinical Microbiology and Infectious Diseases, and held in Rome under the patronage of the Italian government. PMID- 15113334 TI - Fifty Years of the American Society for Artificial Internal Organs: growing up with artificial organs. PMID- 15113335 TI - The role of federal programs in the transfer of biomedical technology. PMID- 15113336 TI - The artificial organ's last frontier. PMID- 15113337 TI - Myths and truths of pulsatile and nonpulsatile perfusion during acute and chronic cardiac support. PMID- 15113338 TI - Treatment of full-thickness skin defect with concomitant grafting of 6-fold extended mesh auto-skin and allogeneic cultured dermal substitute. AB - The aim of this clinical study was to evaluate an allogeneic cultured dermal substitute (CDS) as a biological dressing for highly extended mesh auto-skin grafting. The subjects were five patients with extensive deep burn wounds. Allogeneic CDS was prepared by seeding fibroblasts on a spongy matrix of hyaluronic acid and atelo-collagen. Six-fold extended auto-skin graft was applied to the debrided wound, on which allogeneic CDS was placed. A conventional ointment-gauze dressing was used to protect the CDS. The CDS was applied repeatedly at intervals of 5-7 days. In all cases, the wounds were closed by successful take of mesh auto-skin graft and prompt epithelization from the grafted skin. The skin on the grafted area had a cicatrix appearance, but was soft and thin, maintaining good quality. The application of 6-fold extended auto skin graft in conjunction with allogeneic CDS is an effective method for treatment of extensive severe burn wounds. PMID- 15113339 TI - Automatic system for noninvasive blood pressure determination in rotary pump recipients. AB - In patients with implanted rotary pumps, the arterial pressure pulsatility is usually far lower than in normal individuals. Depending on the remaining degree of pulsatility, cuff-based systems such as the classical Riva-Rocci-determination of arterial blood pressure and correlated sounds or pressure measurements based on cuffpressure oscillations become inaccurate or even impossible. Therefore, a system was developed which evaluates the flow in the radial artery using an ultrasound wristwatch sensor, and this additional information is used for pressure determination. A computerized data acquisition and cuff-control system based on a PC using Matlab software, a wristwatch ultrasound device, and a compressor-driven pressure cuff was set up. The cuff was controlled for automatic inflation and deflation cycles. Cuff pressure and arterial flow was recorded. Several algorithm strategies were developed, which gave data for systolic blood pressure and heart rate together with a reliability index for data quality. Finally, the new algorithms were implemented in a microcontroller system. Comparisons with invasive measurements showed excellent correlation with systolic blood pressure (mean deltaP -0.3 mm Hg, n = 28). During exercise of rotary pump patients and therefore enhanced pulsatility the difference from manual evaluation was -2.1 mm Hg (n = 18). In conclusion, adaptation of the classical cuff-pressure method with ultrasound evaluation of peripheral flow allows reliable determination of blood pressure in patients with low pulsatility resulting from implanted rotary cardiac assist pumps. By development of a wristwatch sensor and an automatic control system a robust method for daily use could be developed. PMID- 15113340 TI - A comparative study between flow visualization and computational fluid dynamic analysis for the sun medical centrifugal blood pump. AB - Flow visualization experiments and computational fluid dynamic (CFD) analyses were performed and the results were compared to clarify the detailed fluid dynamic characteristics for the prototype design of a centrifugal pump, namely, an implantable ventricular assist system from Sun Medical, whose hemocompatibility was previously demonstrated in a series of animal experiments. The flow visualization was conducted with particle tracking velocimetry, and the CFD analysis was performed with STAR-CD software. The findings were as follows: (1). There were no flow separations around the curved open impeller. (2). Antithrombogenic design concepts for the inducer and the vane-shaft clearance were effective in producing axial velocity along the shaft surface and generat ing suitable shear rates against the stationary fluid. (3). Unsteady vortex shedding in the outlet, which adversely affected the fluid dynamic efficiency, was observed clearly by flow visualization. Comparison of velocity distribution measured by flow visualization and CFD analysis showed reasonably good correlation. Our findings indicate that the impeller is suitable for an implantable artificial heart. The techniques of flow visualization and CFD analysis are complementary evaluation tools in research and development efforts. PMID- 15113341 TI - The power-law mathematical model for blood damage prediction: analytical developments and physical inconsistencies. AB - Blood trauma caused by medical devices is a major concern. Complications following the implantation/application of devices such as prosthetic heart valves, cannulae, blood pumps, tubing, and throttles lead to sublethal and lethal damage to platelets and erythrocytes. This damage is provided by the alterations in fluid dynamics, providing a mechanical load on the blood corpuscle's membrane by means of the shear stress. An appropriate quantification of the shear-induced hemolysis of artificial organs is thought to be useful in the design and development of such devices in order to minimize device-induced blood trauma. To date, a power-law mathematical relationship using the time of exposure of a blood corpuscle to a certain mechanical load and the shear stress itself (derived under the peculiar condition of uniform shear stress) has served as a basic model for the estimation of the damage to blood, investigated by means of numerical and/or experimental fluid dynamical techniques. The aim of the present article is to highlight the effect of a time-varying mechanical loading acting on blood cells based on the usual power-law model; furthermore, the effect of the loading history of a blood particle is discussed, showing how the past history of the shear acting on a blood corpuscle is not taken into account, as researchers have done until now. The need for a reassessment of the power-law model for potential blood trauma assessment is discussed by using a mathematical formulation based on the hypotheses of the existence of damage accumulation for blood with respect to time and with respect to shear stress, to be applied in complex flow fields such as the ones established in the presence of artificial organs. PMID- 15113342 TI - Alginate-encapsulated human hepatoblastoma cells in an extracorporeal perfusion system improve some systemic parameters of liver failure in a xenogeneic model. AB - Previous studies have demonstrated that alginate encapsulation of proliferating hepatocyte-derived cell lines (e.g., HepG2 cells) enhances the expression of differentiated hepatocyte function compared with conventional monolayer culture. Furthermore, such capsules have the advantage of cryopreservability, and can be readily manipulated, e.g., for the charging of extracorporeal devices. We utilize a rabbit model of acute liver failure caused by acetaminophen administration to rabbits pretreated to enhance cytochrome p450 enzyme activity, and demonstrate that encapsulated HepG2 cells, in an extracorporeal chamber, perfused by rabbit plasma separated on-line at a rate of 2-5 mL/min, and perfused over cells at 40 60 mL/min, improve systemic parameters of liver failure (diastolic blood pressure and transjugular venous oxygen saturation). Such encapsulated cells have the potential to be developed for extracorporeal liver support systems for acute liver failure. PMID- 15113343 TI - Clinical experience with molecular adsorbent recirculating system (MARS) in patients with drug-induced liver failure. AB - The molecular adsorbent recirculating system (MARS) is a novel extracorporeal technique for liver support. We report the clinical results in a group of fourteen patients with drug-induced liver failure. Fourteen patients, aged 22-83 years, with acute or subacute liver failure [mean Child-Turcotte-Pugh (CTP) score 11 (range 8-15)] due to the intake of various drugs (diet pill overdose-2; Chinese traditional medicine (CTM)-4; antibiotic, paracetamol, tuberculostatic, or vasodilator abuse-8) were treated with one to seven sessions of MARS. Beneficial effects such as the improvement of encephalopathy and prothrombin activity, as well as a reduction of bilirubin and ammonia were recorded during MARS treatments. Thirteen out of fourteen patients survived the hospitalization (93%), and two of the discharged patients died during the follow-up of 6-12 months. The overall survival rate was about 79%. MARS therapy can contribute to the improved treatment of drug-induced liver failure patients. PMID- 15113344 TI - Development of a novel polyimide hollow-fiber oxygenator. AB - We have developed a membrane oxygenator using a novel asymmetric polyimide hollow fiber. The hollow fibers are prepared using a dry/wet phase-inversion process. The gas transfer rates of O(2) and CO(2) through the hollow fibers are investigated in gas-gas and gas-liquid systems. The polyimide hollow fiber has an asymmetric structure characterized by the presence of macrovoids, and the outer diameter of the hollow fiber is 330 microm. It is found that the polyimide hollow fiber oxygenator can enhance the gas transfer rates of O(2) and CO(2), and that the hollow fiber provides excellent blood compatibility in vitro and in vivo. PMID- 15113345 TI - Testing of extracorporeal membrane oxygenation circuit related hemolysis using long-term stored packed red cells and fresh frozen plasma. AB - BACKGROUND: The resistance of blood used in these studies to hemolysis differs markedly from that used in neonatal extracorporeal circulation under clinical circumstances. In this study, the possibility of using expired packed red cells to determine hemolysis caused by mechanical and/or environmental factors was investigated. METHODS: Packed red blood cells stored for 42 days were mixed with fresh frozen plasma and the resultant mixture was divided into three groups, two study groups and a control. For the study groups, two different centrifugal pump heads (Medtronic BP 50 and Jostra RF 32) were used in an extracorporeal membrane oxygenation (ECMO) circuit. Free hemoglobin, lactate dehydrogenase, lactic acid, pH, potassium, and glucose were investigated at various time intervals. RESULTS: Hemolysis did not differ between the groups. Free hemoglobin increased in all groups after 12 h. Lactic acid increased linearly in all groups up to 12 h. Glucose and pH decreased steadily in all groups. Hemolysis created during mock ECMO did not differ between the circuits using the two different pump heads noted. CONCLUSION: Human donor blood stored up to its expiration date is a feasible medium for mock circulation tests of up to 12 h duration under the circumstances described. PMID- 15113346 TI - Numerical estimation of blood damage in artificial organs. AB - The aim of this study was to determine a method for the numerical estimation of blood damage. Normally, human or animal blood is used for in vitro evaluation of lysis by artificial organs. However, blood has some disadvantages: large biological variability and different initial test conditions lead to nonreproducible test results. For that reason, it would be an advantage to have a numerical method for blood damage estimation. This proposed method is based on the calculation of an integrated hemolysis and platelet lysis index along the path line in the flow field of the artificial organ. The time-dependent shear stress related lysis is based on known experimental data. In order to calibrate these data, the method was first applied to blood circulation in the human body. The results showed that the known data overestimate hemolysis by a factor of approximately 25. Next, the method was applied to a standard Bjork-Shiley valve. The flow through a valve was simulated with the computational fluid dynamics program FLUENT. The calculation of lysis was added into FLUENT and done automatically. The results showed that the Bjork-Shiley valve increased the hemolysis index by 7% if implanted in the human body circulation. PMID- 15113347 TI - Particle image velocimetry for flow analysis in longitudinal planes across a mechanical artificial heart valve. AB - In the last decades a great number of in vitro studies have been conducted to improve the design and to understand the transvalvular flow patterns under steady state and pulsating flow conditions. Steady-state tests are useful for studying the flow established upstream and downstream of the valve prosthesis in different flow conditions and, in particular, at the peak flow rate. In the present study, the particle image velocimetry (PIV) technique is employed to visualize the flow patterns in a precommercial model of a bi-leaflet mechanical heart valve prosthesis in a steady-state flow regime. The use of the PIV technique and a convenient test rig provide good conditions with which to investigate the whole flow field upstream and downstream of the valve. Velocity and vorticity maps are obtained for the flow passing through the prosthesis in different axial planes. A new and simple solution for the measurement test chamber is proposed. This innovative approach of observing the flow in staggered planes (other than diametrical planes) allows a flow analysis to be performed upstream and downstream of the valve in regions near the tube wall where it has the maximum potential for injury of the vessel's inner cell layer. PMID- 15113351 TI - Organ donation and transplantation trends in the USA, 2003. PMID- 15113352 TI - Transplant data: sources, collection, and caveats. AB - By examining the sources, quality and organization of transplant data available, as well as making observations about data reporting patterns and accuracy, we hope to improve understanding of existing results, help researchers with study design and stimulate new exploratory initiatives. The primary data source, collected by the OPTN, has benefited from extensive recent technological advances. Transplant professionals now report patient and donor data more easily, quickly, and accurately, improving data timeliness and precision. Secondary sources may be incorporated, improving the accuracy and expanding the scope of analyses. For example, auxiliary mortality data allows more accurate survival analysis and conclusions regarding the completeness of center-reported post transplant follow-up. Furthermore, such sources enable examination of outcomes not reported by centers, such as mortality after waiting list removal, providing more appropriate comparisons of waiting list and post-transplant mortality. Complex collection and reporting processes require specific analytical methods and may lead to potential pitfalls. Patterns in the timing of reporting adverse events differ from those for 'positive' events, yielding the need for care in choosing cohorts and censor dates to avoid bias. These choices are further complicated by the use of multiple sources of data, with different time lags and reporting patterns. PMID- 15113353 TI - Organ donation and utilization in the USA. AB - The processes leading to donor identification, consent, organ procurement, and allocation continue to dominate debates and efforts in the field of transplantation. A considerable shortage of donors remains while the number of patients needing organ transplantation increases. This article reviews the main trends in organ donation practices and procurement patterns from both deceased and living sources in the USA. Although there have been increases in living donation in recent years, 2002 witnessed a much more modest growth of 1%. Absolute declines in living liver and lung donation were also noted in 2002. In 2002, the number of deceased donors increased by only 1.6% (101 donors). Increased donation from deceased donors provides more organs for transplantation than a comparable increase in living donation, because on average 3.6 organs are recovered from each deceased donor. The total number of organs recovered from deceased donors increased by 2.1% (462 organs). Poor organ quality continued to be the major reason given for nonrecovery of consented organs from deceased donors. The kidney is the organ most likely to be discarded after recovery. Over the past decade the discard rate of recovered kidneys has increased from 6% to 11%. Many of these are expanded criteria donor kidneys. PMID- 15113354 TI - Immunosuppression: practice and trends. AB - Over the past decade, immunosuppression therapy has undergone striking changes in the scale and pace by which new immunosuppressive molecules and antibodies have become incorporated into daily transplant medicine. An organ-by-organ review of data reveals several trends. The highest use of induction therapy (over 70% of patients) was reported for simultaneous pancreas kidney (SPK) and pancreas after kidney (PAK) transplants in 2002; use of induction therapy was less common in liver transplants (only 18%). Corticosteroids served as discharge maintenance immunosuppression in over 87% of the recipients of kidney, SPK, PAK and thoracic transplants, and in over 70% of pancreas transplant alone (PTA) recipients. Corticosteroid use in intestine transplants was reported in 64% of recipients in 2002. A shift in the calcineurin inhibitor used for maintenance immunosuppression from cyclosporine to tacrolimus for the majority of patients had occurred for kidney, PAK, SPK, PTA, liver, lung, and heart-lung by 2001. For heart transplants, cyclosporine remained the calcineurin inhibitor of choice; tacrolimus remained the predominant calcineurin inhibitor agent for intestine (since 1994). Use of antibody treatment for rejection during the first post transplant year for most organs declined. Short-term outcomes have improved, based on the observation that rates of rejection within the first year post transplant have diminished. PMID- 15113355 TI - Pediatric transplantation. AB - Analysis of the OPTN/SRTR database demonstrates that, in 2002, pediatric recipients accounted for 7% of all recipients, while pediatric individuals accounted for 14% of deceased organ donors. For children fortunate enough to receive a transplant, there has been continued improvement in outcomes following all forms of transplantation. Current 1-year graft survival is generally excellent, with survival rates following transplantation in many cases equaling or exceeding those of all other recipients. In renal transplantation, despite excellent early graft survival, there is evidence that long-term graft survival for adolescent recipients is well below that of other recipients. A causative role for noncompliance is possible. While the significant improvements in graft and patient survival are laudable, waiting list mortality remains excessive. Pediatric candidates awaiting liver, intestine, and thoracic transplantation face mortality rates generally greater than those of their adult counterparts. This finding is particularly pronounced in patients aged 5 years and younger. While mortality awaiting transplantation is an important consideration in refining organ allocation strategies, it is important to realize that other issues, in addition to mortality, are critical for children. Consideration of the impact of end-stage organ disease on growth and development is often equally important, both while awaiting and after transplantation. PMID- 15113356 TI - Kidney and pancreas transplantation. AB - Data from the Scientific Registry of Transplant Recipients offer a unique and comprehensive view of US trends in kidney and pancreas waiting list characteristics and outcomes, transplant recipient and donor characteristics, and patient and allograft survival. Important findings from our review of developments during 2002 and the decade's transplantation trends appear below. The kidney waiting list has continued to grow, increasing from 47,830 in 2001 to 50,855 in 2002. This growth has occurred despite the increasing importance of living donor transplantation, which rose from 28% of total kidney transplants in 1993 to 43% in 2002. Policies and procedures to expedite the allocation of expanded criteria donor (ECD) kidneys were developed and implemented during 2002, when 15% of deceased donor transplants were performed with ECD kidneys. Unadjusted 1- and 5-year deceased donor kidney allograft survivals were 81% and 51% for ECD kidney recipients, and 90% and 68% for non-ECD kidney recipients, respectively. Although more patients have been placed on the simultaneous kidney pancreas waiting list, the number of these transplants dropped from a peak of 970 in 1998 to 905 in 2002. This decline may be due to competition for organs from increasing numbers of isolated pancreas and islet transplants. PMID- 15113357 TI - Liver and intestine transplantation. AB - The most significant development in liver transplantation in the USA over the past year was the full implementation of the MELD- and PELD-based allocation policy in March 2002, which shifted emphasis from waiting time within broad medical urgency status to prioritization by risk of waiting list death. The implementation of this system has led to a decrease in pretransplant mortality without increasing post-transplant mortality, despite a higher severity of illness at the time of transplant. The trend over the last few years of rapidly increasing numbers of adult living donor liver transplants was reversed in 2002 by a decline of more than 30% in the number of these procedures. In 2002, a greater percentage of women received livers from living donors (43%) than deceased donors (34%), possibly because of size considerations. From 1993 to 2001, the waiting list increased more than sixfold, from 2902 patients to 18,047 patients. For the first time since 1993, the waiting list size decreased in 2002, dropping 6% to 16,974 candidates. The percentage of temporarily inactive liver candidates also increased from 2001, thus the net decrease in the active waiting list for 2002 was 12%. This may reflect a trend toward less pre-emptive listing practices under MELD. Intestine transplantation remains a low-volume procedure limited to a few transplant centers and is still accompanied by significant pre- and post-transplantation risks. As this procedure matures, its application may increase to include recipients at an earlier stage of their disease with better likelihood of success. PMID- 15113358 TI - Thoracic organ transplantation. AB - This article presents an overview of factors associated with thoracic transplantation outcomes over the past decade and provides valuable information regarding the heart, lung, and heart-lung waiting lists and thoracic organ transplant recipients. Waiting list and post-transplant information is used to assess the importance of patient demographics, risk factors, and primary cardiopulmonary disease on outcomes. The time that the typical listed patient has been waiting for a heart, lung, or heart-lung transplant has markedly increased over the past decade, while the number of transplants performed has declined slightly and survival after transplant has plateaued. Waiting list mortality, however, appears to be declining for each organ and for most diseases and high severity subgroups, perhaps in response to recent changes in organ allocation algorithms. Based on perceived inequity in organ access and in response to a mandate from Health Resources and Services Administration, the lung transplant community is developing a lung allocation system designed to minimize deaths on the waiting list while maximizing the benefit of transplant by incorporating post transplant survival and quality of life into the algorithm. Areas where improved data collection could inform evolving organ allocation and candidate selection policies are emphasized. PMID- 15113359 TI - Analytical approaches for transplant research. AB - It is highly desirable to base decisions designed to improve medical practice or organ allocation policies on the analyses of the most recent data available. Yet there is often a need to balance this desire with the added value of evaluating long-term outcomes (e.g. 5-year mortality rates), which requires the use of data from earlier years. This article explains the methods used by the Scientific Registry of Transplant Recipients in order to achieve these goals simultaneously. The analysis of waiting list and transplant outcomes depends strongly on statistical methods that can combine data from different cohorts of patients that have been followed for different lengths of time. A variety of statistical methods have been designed to address these goals, including the Kaplan-Meier estimator, Cox regression models, and Poisson regression. An in-depth description of the statistical methods used for calculating waiting times associated with the various types of organ transplants is provided. Risk of mortality and graft failure, adjusted analyses, cohort selection, and the many complicating factors surrounding the calculation of follow-up time for various outcomes analyses are also examined. PMID- 15113360 TI - Improving liver allocation: MELD and PELD. AB - On February 27, 2002, the liver allocation system changed from a status-based algorithm to one using a continuous MELD/PELD severity score to prioritize patients on the waiting list. Using data from the Scientific Registry of Transplant Recipients, we examine and discuss several aspects of the new allocation, including the development and evolution of MELD and PELD, the relationship between the two scoring systems, and the resulting effect on access to transplantation and waiting list mortality. Additional considerations, such as regional differences in MELD/PELD at transplantation and the predictive effects of rapidly changing MELD/PELD, are also addressed. Death or removal from the waiting list for being too sick for a transplant has decreased in the MELD/PELD era for both children and adults. Children younger than 2 years, however, still have a considerably higher rate of death on the waiting list than adults. A limited definition of ECD livers suggests that they are used more frequently for patients with lower MELD scores. PMID- 15113361 TI - Review article: Osteoporosis and inflammatory bowel disease. AB - Studies using dual-energy X-ray absorptiometry have suggested a high prevalence of osteoporosis in inflammatory bowel disease. However, population-based data on fracture incidence suggest only a small increased risk of fracture amongst patients with inflammatory bowel disease compared with the general population. Therefore, it would be helpful to identify patients with inflammatory bowel disease at particularly high risk for fracture so that these risks might be modified or interventions might be undertaken. The data on calcium intake as a predictor of bone mineral density are conflicting. Although there are data suggesting that a one-time survey to determine current calcium intake will not help to predict bone mineral density in inflammatory bowel disease, persistently reduced calcium intake does appear to lead to lower bone mineral density. In the general population, body mass is strongly correlated with bone mineral density, which also appears to be true in Crohn's disease. Hence, subjects with inflammatory bowel disease and considerable weight loss, or who are obviously malnourished, could be considered for bone mineral density testing, and the finding of a low bone mineral density would suggest the need for more aggressive nutritional support. Although vitamin D is undoubtedly important in bone health, vitamin D intake and serum vitamin D levels do not correlate well with bone mineral density. Sex hormone deficiency can also adversely affect bone health, although a well-developed strategy for sex hormone measurements in patients with inflammatory bowel disease remains to be established. Ultimately, the determination of genetic mutations that accurately predict fracture susceptibility may be the best hope for developing a simplified strategy for managing bone health in inflammatory bowel disease. The therapy of osteoporosis in inflammatory bowel disease has been adapted from other osteoporosis settings, such as post-menopausal or corticosteroid-induced osteoporosis. To date, there remains no therapy proven to be efficacious in inflammatory bowel disease-related osteoporosis; however, calcium and vitamin D supplementation and bisphosphonates have their roles. PMID- 15113362 TI - Review article: The medical treatment of Crohn's perianal fistulas. AB - Perianal fistulas are a frequent manifestation of Crohn's disease. The correct application of the newer diagnostic and therapeutic agents for treating perianal Crohn's disease are beginning to be better defined. In general, a combined medical and surgical approach is preferred. The perianal disease process should first be fully delineated with endoscopy and either MRI or EUS before treatment is begun. Patients are then stratified into one of three groups: simple fistulas and no proctitis; simple fistulas and concomitant proctitis; and complex fistulas. Patients with simple fistulas and no proctitis can be treated medically with a combination of antibiotics and an immunosuppressive agent (azathioprine or mercaptopurine). Patients with simple fistulas and concomitant proctitis should have infliximab added to their treatment plan. Complex fistulas require surgical intervention first prior to medical treatment. A combination of antibiotics, immunosuppressive therapy and infliximab are then initiated to facilitate fistula healing. PMID- 15113363 TI - Review article: The rationale for antidepressant therapy in functional gastrointestinal disorders. AB - The rationale for antidepressants in the treatment of functional gastrointestinal disorders (FGDs) has been the subject of much interest. However, because of our incomplete understanding of FGDs, this rationale remains unclear. A key point is whether the high degree of psychiatric co-morbidity associated with FGDs (40-90%) represents a shared pathophysiology or the ascertainment bias of tertiary referral patients. Our aims were four-fold: (i) to review the current rationale for antidepressant therapy in FGDs; (ii) to review the studies comparing the characteristics of FGDs with both organic gastrointestinal disease and psychiatric disorders; (iii) to propose a model of FGDs which explains the high psychiatric co-morbidity; (iv) to compare the treatment regimes and effectiveness of antidepressants in FGDs and psychiatric illnesses. The review highlights two important observations. Firstly, the characteristics of FGDs are similar to those of affective disorders and dissimilar to those of organic disease. Secondly, although antidepressants benefit FGD sufferers, their benefits in psychiatric illnesses are greater. We conclude that, in view of the degree of similarity between FGDs and affective disorders, FGDs could be considered as affective disorders in their own right and, if the prescription of antidepressants conformed to their use in affective disorders, FGD morbidity would be reduced. PMID- 15113364 TI - Empirical use of antisecretory drug therapy delays diagnosis of upper gastrointestinal adenocarcinoma but does not effect outcome. AB - BACKGROUND: Upper gastrointestinal cancer carries a poor prognosis. Although the incidence of gastric adenocarcinoma is falling, oesophageal adenocarcinoma is increasing. This has been attributed to an increasing prevalence of gastro oesophageal reflux disease, commonly treated empirically in primary care with antisecretory drugs. Treatment has been associated with delayed diagnosis but it is unclear if this influences prognosis. AIMS: To ascertain the effect of antisecretory drugs on time to diagnosis, symptoms, tumour stage and outcome. METHODS: A retrospective cohort study of primary care records for 747 patients diagnosed with upper gastrointestinal adenocarcinoma at South Tees NHS Trust between 1991 and 2001. RESULTS: Mean time from the onset of symptoms to diagnosis was 30 weeks. Mean and median times at the primary care stage were longer than at the hospital stage for both oesophageal and gastric cancer (P < 0.0001). Patients with benign symptoms prescribed antisecretory drugs were referred later than those not on antisecretory drugs (P < 0.0001), as were patients with alarm symptoms (P = 0.0008). Prior use of antisecretory drugs delayed diagnosis by 17.6 weeks (mean) but had no effect on tumour stage at diagnosis or survival. CONCLUSION: Prior antisecretory drug therapy was associated with delayed diagnosis of upper gastrointestinal adenocarcinoma irrespective of presenting symptoms. Concerns that delays might adversely affect tumour stage or long-term survival were not substantiated. PMID- 15113365 TI - Gastric ulcer and malignancy--is there a need for follow-up endoscopy? PMID- 15113366 TI - Effectiveness and pharmaceutical cost of sequential treatment for Helicobacter pylori in patients with non-ulcer dyspepsia. AB - BACKGROUND: A novel 10-day sequential treatment regimen recently achieved a significantly higher eradication rate than standard 7-day therapy in both peptic ulcer disease and non-ulcer dyspepsia. Its higher performance has recently been confirmed using a halved clarithromycin dose in peptic ulcer disease. AIMS: To evaluate whether an acceptable eradication rate could also be obtained by halving the clarithromycin dose in dyspeptic patients and to assess the role of possible factors affecting the outcome of therapy. METHODS: In a prospective, open-label study, 162 patients with non-ulcer dyspepsia and Helicobacter pylori infection, assessed by rapid urease test and histology, were enrolled. Patients were randomized to receive either 10-day sequential therapy, comprising rabeprazole 20 mg b.d. plus amoxicillin 1 g b.d. for the first 5 days, followed by rabeprazole 20 mg b.d., clarithromycin 250 mg b.d. and tinidazole 500 mg b.d. for the remaining 5 days (low-dose therapy), or a similar schedule with clarithromycin 500 mg b.d. (high-dose therapy). Four to six weeks after therapy, H. pylori eradication was assessed by endoscopy/histology. RESULTS: A similar H. pylori eradication rate was observed following low- and high-dose regimens for both per protocol (94% vs. 95%; P = N.S.) and intention-to-treat (93% vs. 94%; P = N.S.) analyses. No major side-effects were reported. Halving the clarithromycin dose leads to a per patient saving in pharmaceutical costs of 24.6 euros. None of the variables examined affected the effectiveness of eradication of the sequential regimen. CONCLUSION: A reduction of the clarithromycin dose does not affect H. pylori eradication with the sequential regimen in non-ulcer dyspepsia and affords lower costs. PMID- 15113367 TI - Acute renal failure after transarterial chemoembolization for hepatocellular carcinoma: a retrospective study of the incidence, risk factors, clinical course and long-term outcome. AB - BACKGROUND: Transarterial chemoembolization is effective for hepatocellular carcinoma. Acute renal failure may occur after transarterial chemoembolization because of radiocontrast agent, but its clinical aspects are unknown. AIM: To investigate the incidence, risk factors and outcome of acute renal failure, defined as increase of serum creatinine > 1.5 mg/dL, after transarterial chemoembolization. METHODS: A total of 235 hepatocellular carcinoma patients with 843 transarterial chemoembolization treatment sessions were analysed. RESULTS: Acute renal failure developed in 56 (23.8%) patients and the estimated risk of developing acute renal failure was 6.6% in each treatment session. Comparison between the episodes of transarterial chemoembolization with and without acute renal failure by using the generalized estimating equation disclosed that Child Pugh class B (odds ratio: 2.6, P = 0.007) and treatment session (odds ratio: 1.3; P < 0.0001) were independent risk factors of acute renal failure. Twenty-seven patients had prolonged renal function impairment. Multivariate analysis by generalized estimating equation showed that Child-Pugh class B (odds ratio: 4.3, P = 0.0004) and diabetes mellitus (odds ratio: 5.2, P < 0.0001) were linked with prolonged acute renal failure, which independently predicted a decreased survival (relative risk: 2.3, P = 0.002). CONCLUSIONS: Acute renal failure after transarterial chemoembolization appears to be dose-related and is associated with the severity of cirrhosis. Patients with diabetes mellitus or Child-Pugh class B more frequently develop prolonged acute renal failure, which in turn is a poor prognostic predictor. PMID- 15113368 TI - Primary Helicobacter pylori resistance to metronidazole and clarithromycin in the Finnish population. AB - AIM: To systematically determine Helicobacter pylori primary antimicrobial resistance in Finland and the associated demographic and clinical features. METHODS: A total of 342 adult patients referred for gastroscopy at 23 centres in different parts of Finland and positive for the rapid biopsy urease test were recruited. Clinical and demographic data were collected via a structured questionnaire. Patients with positive H. pylori culture and successful antibiotic sensitivity determination by the E-test method (n = 292) were included in the present analysis. RESULTS: The study population consisted of 134 men and 158 women, mean age 56 years (95% CI, 55-58 years). Resistance to metronidazole was 38% (110 of 292) and to clarithromycin 2% (seven of 292). Resistance to metronidazole was higher in women than in men (48% vs. 25%, P < 0.001). Previous use of antibiotics for gynaecological infections predicted metronidazole resistance (P = 0.01), and previous use of antibiotics for respiratory (P = 0.02) and dental infections (P = 0.02) the clarithromycin resistance. We observed no major geographical variations in metronidazole resistance. CONCLUSIONS: The primary metronidazole resistance of H. pylori was 38% and was common in women previously treated for gynaecological infections. Primary clarithromycin resistance was uncommon (2%) and may associate with previous dental and respiratory infections. PMID- 15113369 TI - Infliximab downregulates basic fibroblast growth factor and vascular endothelial growth factor in Crohn's disease patients. AB - BACKGROUND: The use of infliximab in the treatment of Crohn's disease patients with symptomatic stenosis is controversial. AIM: To explore the influence of this agent on intestinal fibrogenesis by measuring in infliximab-treated Crohn's disease patients the serum levels of basic fibroblast growth factor and vascular endothelial growth factor, two factors known to be involved in the process of intestinal wound healing and fibrosis in this condition. METHODS: Serum levels of basic fibroblast growth factor and vascular endothelial growth factor were determined by enzyme-linked immunosorbent assay in 22 patients with steroid refractory or fistulizing Crohn's disease before, during (2 weeks) and after 12 weeks of treatment with infliximab, administered at week 0, 2 and 6 in a dose of 5 mg/kg. RESULTS: A substantial improvement in 19 of the 22 Crohn's disease patients was accompanied by a rapid and durable reduction in basic fibroblast growth factor and vascular endothelial growth factor serum levels. CONCLUSIONS: The action of infliximab in reducing serum basic fibroblast growth factor and vascular endothelial growth factor would seem to suggest a role of this agent in down-regulating the process of intestinal fibrogenesis in Crohn's disease. PMID- 15113370 TI - Incidence of a clinical diagnosis of the irritable bowel syndrome in a United States population. AB - BACKGROUND: The incidence of irritable bowel syndrome is uncertain. We aimed to determine the incidence of clinically diagnosed irritable bowel syndrome in the community. METHODS: Using the Rochester Epidemiology Project, all diagnoses of irritable bowel syndrome made among adult residents of Olmsted County, Minnesota, over a 3-year period were identified. The complete medical records of a random sample of the potential subjects were reviewed for the 10 years prior to the irritable bowel syndrome diagnosis and any patient who had received a previous diagnosis of irritable bowel syndrome was excluded (prevalent cases). RESULTS: The diagnostic index listed 1245 possible irritable bowel syndrome patients; 416 patient charts were reviewed and, of these, 149 were physician diagnosed incident cases of irritable bowel syndrome. The age- and sex-adjusted incidence rate was 196 per 100,000 person-years and increased with age (P = 0.006). The age-adjusted annual incidence per 100,000 in women was higher than in men: 238 vs. 141 (ratio 3:2; P = 0.005). The overall symptom frequency at the time of diagnosis was abdominal pain (73%), diarrhoea (41%) and constipation (16%). CONCLUSIONS: The incidence of a clinical diagnosis of irritable bowel syndrome in adults was estimated to be two per 1000 per year, increased with age and was higher in women than men. As many people with irritable bowel syndrome do not seek care, the true incidence of irritable bowel syndrome is likely to be higher. PMID- 15113371 TI - Sirolimus-induced hyperlipidaemia in liver transplant recipients is not dose dependent. AB - BACKGROUND: Sirolimus is a potent immunosuppressive medication that acts by inhibiting T-cell proliferation. It has been used in kidney transplantation because of its lack of nephrotoxicity. It is now being investigated in liver transplantation, but there are concerns about safety and long-term side effects such as dyslipidaemia. Hypertriglyceridaemia is a common adverse event seen with sirolimus use, and often does not respond to dose reduction or anti-lipemic drugs. METHOD: We report six patients who have developed significant hyperlipidaemia while receiving sirolimus, in spite of therapeutic trough levels. CONCLUSION: All six patients showed either resolution or improvement in lipid levels with discontinuation of sirolimus. PMID- 15113372 TI - XXXVI Nordic Congress of Ophthalmology, 16-20 June, 2004, Malmo, Sweden. Abstracts. PMID- 15113373 TI - Immune modulation with dendritic cells. PMID- 15113374 TI - Low-level viraemia of hepatitis B virus in an anti-HBc- and anti-HBs-positive blood donor. AB - summary In many countries, screening of hepatitis B virus (HBV) in blood donors is limited to HBsAg testing. However, if anti-HBc testing and sensitive HBV nucleic acid amplification testing (NAT) for routine screening are not prescribed, HBV viraemia might remain unrecognized. A clinically inconspicuous HBsAg-negative 35-year-old female blood donor was detected with anti-HBc antibodies following the introduction of anti-HBc screening of donors. Based on her history, she had seroconverted to anti-HBs positive (titre >7000 IU/L) after vaccination. Blood donations were routinely tested HBV-DNA negative by minipool NAT. The individual donor samples were reinvestigated by an ultrasensitive NAT with a lower detection limit of 3.8 IU/mL. Intermittent HBV viraemia was detected over a 7-year period from this donor, with a concentration ranging from 8 to 260 IU/mL. In the subsequent donor-directed lookback study, no post-transfusion hepatitis was detected. Low-level HBV viraemia in simultaneous anti-HBc- and anti HBs-positive blood donors could only be identified with enhanced sensitivity individual polymerase chain reaction assays and is not detectable by pool HBV NAT. PMID- 15113375 TI - Comparative analysis of national regulations concerning blood safety across Europe. AB - In October 2001, representatives of 17 European countries (Albania, Bosnia Herzegovina, Bulgaria, Croatia, Czech Republic, Federal Republic of Yugoslavia, Finland, France, Germany, Greece, Italy, Macedonia, Romania, Slovenia, Spain, Turkey and UK) met in Sarajevo at a course organized by the European School of Transfusion Medicine to discuss their countries' regulations concerning different aspects of the safety of blood transfusion. Results are summarized in tables to facilitate comparisons. Most countries (13/17) have specific transfusion laws and 9/17 have hospital-based systems as opposed to national organizations. Quality assurance is common among investigated countries (14/17). Voluntary associations are responsible for donor promotion in the majority of countries (13/17). Exclusively, voluntary non-remunerated donors are found in 5/17 countries, whereas in the remaining ones, incentives, family replacement and remuneration are mechanisms stimulating blood donation. Medical doctors using official selection criteria are checking donor suitability in virtually all countries, which also perform main microbiological testing. Regulations on good clinical use of blood and derivatives are present in most countries but applied only in some. Although the data presented need to be interpreted with some caution, this preliminary analysis shows that, although some significant differences still exist, the majority of countries studied are moving in the same direction to ensure safety of their blood supply. PMID- 15113376 TI - Current performance of patient sample collection in the UK. AB - Collection of the patient sample for pretransfusion testing begins a complex chain of events in the transfusion process. Hospitals in England and North Wales were surveyed to compare local policies against recommendations of the British Committee for Standards in Haematology (BCSH). Hospitals also measured the frequency of rejected and miscollected samples [designated as wrong blood in tube (WBIT)]. 185 of 360 (51.4%) hospitals returned questionnaires and 182 of 185 (98%) hospitals reported that a policy for sample collection existed. Apart from frequent omission of the gender of the patient, there was 96% compliance with all mandatory identifiers of the BCSH guidelines. Practice allowing additions or changes to labelling on sample tubes and request forms varied. 3.2% (14 114/445 726) of samples submitted were rejected for various reasons, the most frequent being incomplete or missing information (49.5% of the total rejected samples). The corrected mean frequency for WBIT in the 27 hospitals with one or more observed WBIT was 1 in 1501 samples (95% CI: < or =1129.09 to < or =1872.91), and the median corrected frequency for WBIT was 1 in 1303 samples. This study has identified great variation in the policy and practice for sample collection for pretransfusion testing. Regular tracking of the rates of sample rejection and WBIT could be used to identify poor performance in individual hospitals requiring investigation and action. PMID- 15113377 TI - Autologous transfusion techniques: a systematic review of their efficacy. AB - Shortages of donor blood and fears of transmitted infections have prompted the use of a range of blood-sparing techniques in the peri-operative period. We conducted a systematic review of three techniques that involve the re-infusion of a patient's blood--pre-operative autologous blood deposit (PAD), acute normovolaemic haemodilution (ANH), and cell salvage (CS). We examined the effects of these interventions on the need for peri-operative allogeneic red blood cell transfusion and on clinical outcomes. Controlled clinical studies were identified by computer searches of comprehensive electronic databases and bibliographic searches of published articles. The literature search retrieved a total of 68 randomized trials (RCTs) and 81 controlled observational studies that included data from over 34 000 individuals. In summary, the RCTs found that autologous transfusion techniques consistently reduced the frequency of allogeneic transfusions, with intervention effect sizes ranging from a relative 63% reduction (95% CI 46-74%) with PAD, to 42% (27-53%) with CS and to 31% (16-44%) with ANH. Non-randomized studies reported larger effect sizes than RCTs. PAD increased overall transfusion rates by 30% (95% CI 12-48%) and reduced pre operative haemoglobin levels by an average of 1.23 g dL(-1). Intervention effects were substantially reduced when these techniques were performed under transfusion protocols. Interpretation of the studies was hampered by serious methodological weaknesses, particularly inadequate randomization techniques, unblinded measurements and the subjective nature of the outcome variables. The studies reported few clinical outcome and adverse event data. Previous claims regarding reduced rates of mortality and infection with autologous transfusions were not confirmed. PMID- 15113378 TI - Implementation of concurrent red blood cell and platelet collection by apheresis in a university haemapheresis unit. AB - The present study analyses the number of concurrently collected red blood cell (RBC) units in plateletpheresis donors and the reasons why donors were deferred from multicomponent collection. Donors undergoing concurrent collection of RBCs and platelets (PLTs) were retrospectively evaluated for haemoglobin values and the reasons for deferral over a period of 1 year. A total of 404 RBC units were concurrently collected with PLTs. An average of 1.8 RBC units per year was collected from each donor. The baseline haemoglobin values were almost equal for the RBC donations. An RBC unit was not collected in 190 aphereses. Most frequent reasons for the noncollection of an RBC product were a donation interval of less than 3 months (20.5%), haematoma and blood flow problems (18.9%) and low pre haemoglobin values (17.4%). Donor eligibility has to be taken into account to optimize concurrent RBC collection in plateletpheresis. PMID- 15113379 TI - Molecular evidence of tumour cell removal from salvaged blood after irradiation and leucocyte depletion. AB - Intra-operative autologous blood recovery offers many advantages. However, blood salvage during cancer surgery is of limited use due to the potential presence of circulating tumour cells. It was the aim of this study to show that intra operative salvage blood can be freed of cells and cellular DNA after leucoreduction by filtration and irradiation of washed blood. Known amounts of tissue culture derived from carcinoma, melanoma and osteosarcoma were added to whole blood bags. This mixture was then submitted to washing, leucoreduction and irradiation. Samples were studied stepwise in relation to the integrity and size of DNA by the polymerase chain reaction (PCR). After filtration and irradiation, PCR targeting the beta-globin gene (268 bp amplicon) was negative. Our results were corroborated by studying plasma samples added with tumoural cells. Using PCR methodology, we showed the absence of DNA from cells in experimentally contaminated blood and plasma bags after filtration and irradiation. This experimental study is an effort to ensure the safety of intra-operative autologous transfusion. PMID- 15113380 TI - Implementation of a strategy to prevent TRALI in a regional blood centre. AB - Transfusion-related acute lung injury (TRALI) can be a life-threatening complication of transfusion and it is probably underdiagnosed. Human leucocyte antigen (HLA) and granulocyte antibodies are thought to play a major role, but preventive measures are difficult to implement. In our regional blood centre, we implemented a preventive strategy avoiding donor deferral. Previously, pregnant apheresis donors were screened for HLA antibodies, and those with positive results were assigned to a plasma-only protocol. Plasma from these donors and from all previously pregnant whole blood donors was diverted for protein fractionation. Plasma-poor red blood cells (in additive solution, buffy coat removed) and platelets (pools with additive solution) were prepared. Prestorage leucodepletion was also applied. We found HLA antibodies in 18.1% of previously pregnant apheresis donors, and our strategy caused a 6.0% loss of apheresis platelets, a 4.8% increase of apheresis fresh frozen plasma (FFP) and a 7.8% loss of transfusable apheresis FFP. The effect on FFP from whole blood donors could be compensated. The platelet preparation method reduced the mean volume of plasma from each donor to 24.4 mL. Fifteen months after the start of our strategy, no cases of TRALI have been reported. Our experience shows that a practical strategy to prevent TRALI is feasible. PMID- 15113381 TI - Pharmacokinetics and safety of recombinant anti-RhD in healthy RhD-negative male volunteers. AB - In this first-in-man study, we assessed the pharmacokinetics, safety and tolerability of MonoRho, a human recombinant monoclonal anti-RhD immunoglobulin G1 (IgG1) antibody. Eighteen RhD-negative healthy male volunteers were randomized in two groups to receive a single administration of 300 micro g of MonoRho either intravenously or intramuscularly. There were no symptoms of allergic or anaphylactic type reaction in any subject, and there was no evidence of any MonoRho-related changes in laboratory safety parameters. None of the subjects mounted a detectable immune response to MonoRho. Serum samples were obtained up to 91 days after injection to measure anti-D IgG concentrations by flow cytometry. After intramuscular administration of MonoRho, anti-D IgG concentrations gradually increased reaching peak levels after a mean of 3.4 days. After 3 weeks, the mean anti-D IgG concentrations after intravenous and intramuscular administration became virtually equal to each other and remained so thereafter. In both the treatment groups, the mean elimination half-life was about 18 days and thus similar to that described for plasma-derived anti-D IgG. The bioavailability of MonoRho after intramuscular administration was estimated as 46%. The excellent tolerability and safety of MonoRho as well as its expected elimination half-life supports the continued clinical development of this compound. PMID- 15113382 TI - A novel blood group B subgroup: serological and genetic studies. AB - A discrepancy in the ABO blood groups between a newborn child and her parents was identified. Serological and DNA investigative techniques were performed. A weak variant of B (B(w)) was detected on the erythrocytes of the child, her grandmother and great-uncle. Adsorption-elution studies showed that their erythrocytes adsorb and yield anti-B on elution. The B(w) antigenic strength of the A(1)B(w) cells of her mother and maternal aunt was reduced when compared to that of the A(2)B(w) from another family member. Only one of 15 different anti-B sera agglutinated the A(1)B(w) erythrocytes. Agglutinin anti-B that reacted strongly with normal B erythrocytes and did not agglutinate the B(w) cells, was found in the sera of the A(1)B(w) individuals. The B(w) serum glycosyltransferase could not convert O cells into B cells and no B substance was found in saliva. All family members with the B(w)/AB(w) phenotypes were heterozygous for a B allele and DNA sequencing revealed a novel missense mutation in exon 7 of the B allele (556A > G), resulting in M186V. This substitution changes a highly conserved region of the enzyme, proposed to be a disordered loop near the enzyme cleft, and is expected to diminish the enzyme's activity, leading to this B(w) phenotype. PMID- 15113383 TI - Unconventional diagnosis of Normandy-type von Willebrand's disease in a blood donor. AB - We report the case of a long-standing female blood donor whose blood donation was processed for cryoprecipitate. The cryoprecipitate unit was chosen at random for FVIII:C estimation as part of the quality control, and a low FVIII:C level was identified. The cause of this was subsequently shown to be the Normandy variant of type-2 von Willebrand's disease due to a homozygous Arg854Gln mutation in the von Willebrand factor gene. PMID- 15113384 TI - A new hybrid RHD-positive, D antigen-negative allele. PMID- 15113385 TI - Variations of factor VIII:C plasma levels with respect to the blood group ABO. PMID- 15113386 TI - Myocardial ischaemia was not precipitated by acute normovolaemic haemodilution alone. PMID- 15113389 TI - Progression and regression in renal vascular and glomerular fibrosis. AB - End-stage renal disease (ESRD) is characterized by the development of fibrotic lesions in the glomerular, interstitial and vascular compartments. Renal fibrogenesis, a common complication of diabetes and hypertension, is a complex dynamic process involving several players such as inflammatory agents, cytokines, vasoactive agents and enzymes participating in extracellular matrix assembly, anchoring or degradation. The only available treatment today against chronic renal failure is dialysis or kidney transplantation, making thus ESRD one of the most expensive diseases to treat on a per-patient basis. An emerging challenge for clinicians, maybe the nephrologist's Holy Grail in the 21st century, is to stop definitively the decline of renal function and, if possible, to achieve regression of renal fibrosis and restoration of renal structure. Over the last 5 years, different approaches have been tested in experimental models of nephropathy with variable degree of success. In this review, we will focus on the mechanisms of the hypertension-associated fibrosis and the few recent studies that gave promising results for a therapeutic intervention. PMID- 15113390 TI - Expression of somatostatin mRNA and peptides in C-cell tumours of the thyroid gland in Han Wistar rats. AB - C-cell tumours of the thyroid gland are among the most common spontaneous neoplasms of the laboratory rat. With the exception of calcitonin, little attention has been paid to the secretory peptides of C cells during the development of neoplasia. Of these peptides, somatostatin (SS) is of particular interest because it has been shown to have a direct anti-secretory effect on both thyroid follicular and C cells in vitro. In the present study, in situ hybridization and immunohistochemistry were used to investigate the expression of SS mRNA and SS peptides, in normal C cells and a range of spontaneous proliferative C-cell lesions in the Han Wistar rat. It was confirmed that a small minority of C cells in the normal rat thyroid gland produce and store SS peptides; however, approximately half of all C-cell adenomas and C-cell carcinomas stained positively for SS mRNA and peptides. SS expression was also observed in all metastatic deposits of carcinomas in drainage lymph nodes. From these observations, it appears that C-cell tumours are more likely to develop from SS-expressing stem cells, rather than from non-SS-expressing stem cells. In addition, a lack of differentiation of neoplastic C cells, or reversion to more primitive cell types, could account for increased number of cells expressing SS in C-cell tumours relative to the normal C-cell population. Finally, the mean percentage of cells that stained positively for SS mRNA and peptides appeared to be significantly higher in small C-cell tumours, suggesting that SS may have exerted a growth-controlling influence on these lesions. PMID- 15113391 TI - Increased oxidative stress and apoptosis in acute puromycin aminonucleoside nephrosis. AB - Accumulating evidence demonstrates that oxidative stress is one of the underlying mechanisms to induce apoptosis in different biological systems. The aim of this study was to examine the simultaneous presence and correlation between oxidative stress events, apoptosis, apoptosis-associated proteins and monocyte/macrophage infiltration during the course of acute puromycin aminonucleoside nephrosis (PAN). To induce nephrosis, Sprague-Dawley rats were injected intraperitoneally with puromycin aminonucleoside and killed at weeks 1 and 2 of nephrosis. Controls represent animals injected with 0.9% saline solution. Kidney sections were homogenized to measure nitric oxide (NO), malondialdehyde (MDA), reduced glutathione (GSH), superoxide dismutase (SOD) and catalase activities by appropriate enzymatic and biochemical methods. Renal frozen sections were studied for superoxide anion (O(2) (-)) by a histochemical method, for apoptosis by TUNEL (terminal-deoxynucleotidyl-transferase-mediated dUTP- digoxigenin nick end labelling) and for apoptosis-associated protein expression and monocyte/macrophage infiltration by monoclonal antibodies. Increased renal apoptosis, p53, Bax, Bcl-2 accompanied by increased O(2) (-) and NO generation, lipid peroxidation (MDA) and monocyte/macrophage infiltration were found in nephrotic animals. Renal oxidative stress (O(2) (-), NO and MDA) was correlated with apoptosis, p53 expression, monocyte/macrophage cells and proteinuria. Anti oxidant molecules (SOD and GSH) remained unchanged apart from a decreased activity of catalase which correlated with glomerular apoptosis. In conclusion, the close correlation between the presence of apoptosis and oxidative events confirms the role of oxidative stress in the apoptosis observed during PAN. PMID- 15113393 TI - British Society for Matrix Biology Meeting. London, United Kingdom, 18-19 September 2003. Abstracts. PMID- 15113392 TI - Evidence of macrophage and lymphocyte, but not dendritic cell, infiltration in posterior uveal melanomas, whilst cultured uveal melanomas demonstrate pluripotency by expressing CD68 and CD163. AB - Although variation in the level of macrophage infiltration has been reported in uveal melanoma, little is known about the expression of other leucocyte markers. An immuno- histochemistry study of the levels of expression of macrophage and other leucocyte markers, in a series of 10 primary choroidal melanoma biopsies, was undertaken. Biopsies were either fixed immediately in formalin and embedded in paraffin wax or established as short-term cultures. Using single- and double labelling immunohistochemistry, cultured cells and paraffin sections were analysed for a range of melanoma (HMB45, Melan A, S100 and tyrosinase) and immune cell (CD68, CD163, CD45 and CD1a) markers. All samples expressed at least two known melanoma markers. Infiltrating macrophages were present in the majority of sections. When cultured specimens were studied by double-labelling immunofluorescence, uveal melanoma cells were seen to express macrophage markers or have cross-reactivity with related proteins. Expression of the leucocyte antigen CD45 was observed in three tumours but was not present in any cultured cells, whilst the expression of the dendritic cell marker CD1a was absent from all samples. PMID- 15113395 TI - The psychosocial and health care needs of HIV-positive people in the United Kingdom: a review. PMID- 15113396 TI - First isolation and further characterization of enteropathogenic Escherichia coli (EPEC) O157:H45 strains from cattle. AB - BACKGROUND: Enteropathogenic Escherichia coli (EPEC), mainly causing infantile diarrhoea, represents one of at least six different categories of diarrheagenic E. coli with corresponding distinct pathogenic schemes. The mechanism of EPEC pathogenesis is based on the ability to introduce the attaching-and-effacing (A/E) lesions and intimate adherence of bacteria to the intestinal epithelium. The role and the epidemiology of non-traditional enteropathogenic E. coli serogroup strains are not well established. E. coli O157:H45 EPEC strains, however, are described in association with enterocolitis and sporadic diarrhea in human. Moreover, a large outbreak associated with E. coli O157:H45 EPEC was reported in Japan in 1998. During a previous study on the prevalence of E. coli O157 in healthy cattle in Switzerland, E. coli O157:H45 strains originating from 6 fattening cattle and 5 cows were isolated. In this study, phenotypic and genotypic characteristics of these strains are described. Various virulence factors (stx, eae, ehxA, astA, EAF plasmid, bfp) of different categories of pathogenic E. coli were screened by different PCR systems. Moreover, the capability of the strains to adhere to cells was tested on tissue culture cells. RESULTS: All 11 sorbitol-positive E. coli O157:H45 strains tested negative for the Shiga toxin genes (stx), but were positive for eae and were therefore considered as EPEC. All strains harbored eae subtype alpha1. The gene encoding the heat-stable enterotoxin 1 (EAST1) was found in 10 of the 11 strains. None of the strains, however, carried ehx A genes. The capability of the strains to adhere to cells was shown by 10 strains harbouring bfp gene by localized adherence pattern on HEp-2 and Caco-2 cells. CONCLUSION: This study reports the first isolation of typical O157:H45 EPEC strains from cattle. Furthermore, our findings emphasize the fact that E. coli with the O157 antigen are not always STEC but may belong to other pathotypes. Cattle seem also to be a reservoir of O157:H45 EPEC strains, which are described in association with human diseases. Therefore, these strains appear to play a role as food borne pathogens and have to be considered and evaluated in view of food safety aspects. PMID- 15113398 TI - ChromSorter PC: a database of chromosomal regions associated with human prostate cancer. AB - BACKGROUND: Our increasing use of genetic and genomic strategies to understand human prostate cancer means that we need access to simplified and integrated information present in the associated biomedical literature. In particular, microarray gene expression studies and associated genetic mapping studies in prostate cancer would benefit from a generalized understanding of the prior work associated with this disease. This would allow us to focus subsequent laboratory studies to genomic regions already related to prostate cancer by other scientific methods. We have developed a database of prostate cancer related chromosomal information from the existing biomedical literature. The input material was based on a broad literature search with subsequent hand annotation of information relevant to prostate cancer. DESCRIPTION: The database was then analyzed for identifiable trends in the whole scale literature. We have used this database, named ChromSorter PC, to present graphical summaries of chromosomal regions associated with prostate cancer broken down by age, ethnicity and experimental method. In addition we have placed the database information on the human genome using the Generic Genome Browser tool that allows the visualization of the data with respect to user generated datasets. CONCLUSIONS: We have used this database as an additional dataset for the filtering of genes identified through genetics and genomics studies as warranting follow-up validation studies. We would like to make this dataset publicly available for use by other groups. Using the Genome Browser allows for the graphical analysis of the associated data http://www.prostategenomics.org/datamining/chrom-sorter_pc.html. Additional material from the database can be obtained by contacting the authors (mdatta@mcw.edu). PMID- 15113397 TI - Endothelial cells present antigens in vivo. AB - BACKGROUND: Immune recognition of vascular endothelial cells (EC) has been implicated in allograft rejection, protection against pathogens, and lymphocyte recruitment. However, EC pervade nearly all tissues and predominate in none, complicating any direct test of immune recognition. Here, we examined antigen presentation by EC in vivo by testing immune responses against E. coli beta galactosidase (beta-gal) in two lines of transgenic mice that express beta-gal exclusively in their EC. TIE2-lacZ mice express beta-gal in all EC and VWF-lacZ mice express beta-gal in heart and brain microvascular EC. RESULTS: Transgenic and congenic wild type FVB mice immunized with beta-gal expression vector DNA or beta-gal protein generated high titer, high affinity antisera containing comparable levels of antigen-specific IgG1 and IgG2a isotypes, suggesting equivalent activation of T helper cell subsets. The immunized transgenic mice remained healthy, their EC continued to express beta-gal, and their blood vessels showed no histological abnormalities. In response to beta-gal in vitro, CD4+ and CD8+ T cells from immunized transgenic and FVB mice proliferated, expressed CD25, and secreted IFN-gamma. Infection with recombinant vaccinia virus encoding beta gal raised equivalent responses in transgenic and FVB mice. Hearts transplanted from transgenic mice into FVB mice continued to beat and the graft EC continued to express beta-gal. These results suggested immunological ignorance of the transgene encoded EC protein. However, skin transplanted from TIE2-lacZ onto FVB mice lost beta-gal+ EC and the hosts developed beta-gal-specific antisera, demonstrating activation of host immune effector mechanisms. In contrast, skin grafted from TIE2-lacZ onto VWF-lacZ mice retained beta-gal+ EC and no antisera developed, suggesting a tolerant host immune system. CONCLUSION: Resting, beta gal+ EC in transgenic mice tolerize specific lymphocytes that would otherwise respond against beta-gal expressed by EC within transplanted skin. We conclude that EC effectively present intracellular "self" proteins to the immune system. However, antigen presentation by EC does not delete or anergize a large population of specific lymphocytes that respond to the same protein following conventional immunization with protein or expression vector DNA. These results clearly demonstrate striking context sensitivity in the immune recognition of EC, a subtlety that must be better understood in order to treat immune diseases and complications involving the vasculature. PMID- 15113399 TI - Comparing gene discovery from Affymetrix GeneChip microarrays and Clontech PCR select cDNA subtraction: a case study. AB - BACKGROUND: Several high throughput technologies have been employed to identify differentially regulated genes that may be molecular targets for drug discovery. Here we compared the sets of differentially regulated genes discovered using two experimental approaches: a subtracted suppressive hybridization (SSH) cDNA library methodology and Affymetrix GeneChip technology. In this "case study" we explored the transcriptional pattern changes during the in vitro differentiation of human monocytes to myeloid dendritic cells (DC), and evaluated the potential for novel gene discovery using the SSH methodology. RESULTS: The same RNA samples isolated from peripheral blood monocyte precursors and immature DC (iDC) were used for GeneChip microarray probing and SSH cDNA library construction. 10,000 clones from each of the two-way SSH libraries (iDC-monocytes and monocytes-iDC) were picked for sequencing. About 2000 transcripts were identified for each library from 8000 successful sequences. Only 70% to 75% of these transcripts were represented on the U95 series GeneChip microarrays, implying that 25% to 30% of these transcripts might not have been identified in a study based only on GeneChip microarrays. In addition, about 10% of these transcripts appeared to be "novel", although these have not yet been closely examined. Among the transcripts that are also represented on the chips, about a third were concordantly discovered as differentially regulated between iDC and monocytes by GeneChip microarray transcript profiling. The remaining two thirds were either not inferred as differentially regulated from GeneChip microarray data, or were called differentially regulated but in the opposite direction. This underscores the importance both of generating reciprocal pairs of SSH libraries, and of real time RT-PCR confirmation of the results. CONCLUSIONS: This study suggests that SSH could be used as an alternative and complementary transcript profiling tool to GeneChip microarrays, especially in identifying novel genes and transcripts of low abundance. PMID- 15113400 TI - Universal Reference RNA as a standard for microarray experiments. AB - BACKGROUND: Obtaining reliable and reproducible two-color microarray gene expression data is critically important for understanding the biological significance of perturbations made on a cellular system. Microarray design, RNA preparation and labeling, hybridization conditions and data acquisition and analysis are variables difficult to simultaneously control. A useful tool for monitoring and controlling intra- and inter-experimental variation is Universal Reference RNA (URR), developed with the goal of providing hybridization signal at each microarray probe location (spot). Measuring signal at each spot as the ratio of experimental RNA to reference RNA targets, rather than relying on absolute signal intensity, decreases variability by normalizing signal output in any two color hybridization experiment. RESULTS: Human, mouse and rat URR (UHRR, UMRR and URRR, respectively) were prepared from pools of RNA derived from individual cell lines representing different tissues. A variety of microarrays were used to determine percentage of spots hybridizing with URR and producing signal above a user defined threshold (microarray coverage). Microarray coverage was consistently greater than 80% for all arrays tested. We confirmed that individual cell lines contribute their own unique set of genes to URR, arguing for a pool of RNA from several cell lines as a better configuration for URR as opposed to a single cell line source for URR. Microarray coverage comparing two separately prepared batches each of UHRR, UMRR and URRR were highly correlated (Pearson's correlation coefficients of 0.97). CONCLUSION: Results of this study demonstrate that large quantities of pooled RNA from individual cell lines are reproducibly prepared and possess diverse gene representation. This type of reference provides a standard for reducing variation in microarray experiments and allows more reliable comparison of gene expression data within and between experiments and laboratories. PMID- 15113401 TI - The over-representation of binary DNA tracts in seven sequenced chromosomes. AB - BACKGROUND: DNA tracts composed of only two bases are possible in six combinations: A+G (purines, R), C+T (pyrimidines, Y), G+T (Keto, K), A+C (Imino, M), A+T (Weak, W) and G+C (Strong, S). It is long known that all-pyrimidine tracts, complemented by all-purines tracts ("R.Y tracts"), are excessively present in analyzed DNA. We have previously shown that R.Y tracts are in vast excess in yeast promoters, and brought evidence for their role in gene regulation. Here we report the systematic mapping of all six binary combinations on the level of complete sequenced chromosomes, as well as in their different subregions. RESULTS: DNA tracts composed of the above binary base combinations have been mapped in seven sequenced chromosomes: Human chromosomes 21 and 22 (the major contigs); Drosophila melanogaster chr. 2R; Caenorhabditis elegans chr. I; Arabidopsis thaliana chr. II; Saccharomyces cerevisiae chr. IV and M. jannaschii. A huge over-representation, reaching million-folds, has been found for very long tracts of all binary motifs except S, in each of the seven organisms. Long R.Y tracts are the most excessive, except in D. melanogaster, where the K.M motif predominates. S (G, C rich) tracts are in excess mainly in CpG islands; the W motif predominates in bacteria. Many excessively long W tracts are nevertheless found also in the archeon and in the eukaryotes. The survey of complete chromosomes enables us, for the first time, to map systematically the intergenic regions. In human and other chromosomes we find the highest over-representation of the binary DNA tracts in the intergenic regions. These over-representations are only partly explainable by the presence of interspersed elements. CONCLUSIONS: The over-representation of long DNA tracts composed of five of the above motifs is the largest deviation from randomness so far established for DNA, and this in a wide range of eukaryotic and archeal chromosomes. A propensity for ready DNA unwinding is proposed as the functional role, explaining the evolutionary conservation of the huge excesses observed. PMID- 15113402 TI - Improving the statistical detection of regulated genes from microarray data using intensity-based variance estimation. AB - BACKGROUND: Gene microarray technology provides the ability to study the regulation of thousands of genes simultaneously, but its potential is limited without an estimate of the statistical significance of the observed changes in gene expression. Due to the large number of genes being tested and the comparatively small number of array replicates (e.g., N = 3), standard statistical methods such as the Student's t-test fail to produce reliable results. Two other statistical approaches commonly used to improve significance estimates are a penalized t-test and a Z-test using intensity-dependent variance estimates. RESULTS: The performance of these approaches is compared using a dataset of 23 replicates, and a new implementation of the Z-test is introduced that pools together variance estimates of genes with similar minimum intensity. Significance estimates based on 3 replicate arrays are calculated using each statistical technique, and their accuracy is evaluated by comparing them to a reliable estimate based on the remaining 20 replicates. The reproducibility of each test statistic is evaluated by applying it to multiple, independent sets of 3 replicate arrays. Two implementations of a Z-test using intensity-dependent variance produce more reproducible results than two implementations of a penalized t-test. Furthermore, the minimum intensity-based Z-statistic demonstrates higher accuracy and higher or equal precision than all other statistical techniques tested. CONCLUSION: An intensity-based variance estimation technique provides one simple, effective approach that can improve p-value estimates for differentially regulated genes derived from replicated microarray datasets. Implementations of the Z-test algorithms are available at http://vessels.bwh.harvard.edu/software/papers/bmcg2004. PMID- 15113403 TI - Current limitations of SNP data from the public domain for studies of complex disorders: a test for ten candidate genes for obesity and osteoporosis. AB - BACKGROUND: Public SNP databases are frequently used to choose SNPs for candidate genes in the association and linkage studies of complex disorders. However, their utility for such studies of diseases with ethnic-dependent background has never been evaluated. RESULTS: To estimate the accuracy and completeness of SNP public databases, we analyzed the allele frequencies of 41 SNPs in 10 candidate genes for obesity and/or osteoporosis in a large American-Caucasian sample (1,873 individuals from 405 nuclear families) by PCR-invader assay. We compared our results with those from the databases and other published studies. Of the 41 SNPs, 8 were monomorphic in our sample. Twelve were reported for the first time for Caucasians and the other 29 SNPs in our sample essentially confirmed the respective allele frequencies for Caucasians in the databases and previous studies. The comparison of our data with other ethnic groups showed significant differentiation between the three major world ethnic groups at some SNPs (Caucasians and Africans differed at 3 of the 18 shared SNPs, and Caucasians and Asians differed at 13 of the 22 shared SNPs). This genetic differentiation may have an important implication for studying the well-known ethnic differences in the prevalence of obesity and osteoporosis, and complex disorders in general. CONCLUSION: A comparative analysis of the SNP data of the candidate genes obtained in the present study, as well as those retrieved from the public domain, suggests that the databases may currently have serious limitations for studying complex disorders with an ethnic-dependent background due to the incomplete and uneven representation of the candidate SNPs in the databases for the major ethnic groups. This conclusion attests to the imperative necessity of large-scale and accurate characterization of these SNPs in different ethnic groups. PMID- 15113404 TI - The roles of cis-inactivation by Notch ligands and of neuralized during eye and bristle patterning in Drosophila. AB - BACKGROUND: The receptor protein Notch and its ligand Delta are expressed throughout proneural regions yet non-neural precursor cells are defined by Notch activity and neural precursor cells by Notch inactivity. Not even Delta overexpression activates Notch in neural precursor cells. It is possible that future neural cells are protected by cis-inactivation, in which ligands block activation of Notch within the same cell. The Delta-ubiquitin ligase Neuralized has been proposed to antagonize cis-inactivation, favoring Notch activation. Cis inactivation and role of Neuralized have not yet been studied in tissues where neural precursor cells are resistant to nearby Delta, however, such as the R8 cells of the eye or the bristle precursor cells of the epidermis. RESULTS: Overexpressed ligands could block Notch signal transduction cell-autonomously in non-neural cells of the epidermis and retina, but did not activate Notch nonautonomously in neural cells. High ligand expression levels were required for cis-inactivation, and Serrate was more effective than Delta, although Delta is the ligand normally regulating neural specification. Differences between Serrate and Delta depended on the extracellular domains of the respective proteins. Neuralized was found to act cell nonautonomously in signal-sending cells during eye development, inconsistent with the view that Neuralized antagonizes cis inactivation in non-neural cells. CONCLUSIONS: Delta and Neuralized contribute cell nonautonomously to Notch signaling in neurogenesis, and the model that Neuralized antagonizes cis-inactivation to permit Notch activity and specification of non-neural cells is refuted. The molecular mechanism rendering Notch insensitive to paracrine activation in neural precursor cells remains uncertain. PMID- 15113405 TI - Defining transcriptional networks through integrative modeling of mRNA expression and transcription factor binding data. AB - BACKGROUND: Functional genomics studies are yielding information about regulatory processes in the cell at an unprecedented scale. In the yeast S. cerevisiae, DNA microarrays have not only been used to measure the mRNA abundance for all genes under a variety of conditions but also to determine the occupancy of all promoter regions by a large number of transcription factors. The challenge is to extract useful information about the global regulatory network from these data. RESULTS: We present MA-Networker, an algorithm that combines microarray data for mRNA expression and transcription factor occupancy to define the regulatory network of the cell. Multivariate regression analysis is used to infer the activity of each transcription factor, and the correlation across different conditions between this activity and the mRNA expression of a gene is interpreted as regulatory coupling strength. Applying our method to S. cerevisiae, we find that, on average, 58% of the genes whose promoter region is bound by a transcription factor are true regulatory targets. These results are validated by an analysis of enrichment for functional annotation, response for transcription factor deletion, and over-representation of cis-regulatory motifs. We are able to assign directionality to transcription factors that control divergently transcribed genes sharing the same promoter region. Finally, we identify an intrinsic limitation of transcription factor deletion experiments related to the combinatorial nature of transcriptional control, to which our approach provides an alternative. CONCLUSION: Our reliable classification of ChIP positives into functional and non-functional TF targets based on their expression pattern across a wide range of conditions provides a starting point for identifying the unknown sequence features in non-coding DNA that directly or indirectly determine the context dependence of transcription factor action. Complete analysis results are available for browsing or download at http://bussemaker.bio.columbia.edu/papers/MA-Networker/. PMID- 15113406 TI - Frequent occurrence of recognition site-like sequences in the restriction endonucleases. AB - BACKGROUND: There are two different theories about the development of the genetic code. Woese suggested that it was developed in connection with the amino acid repertoire, while Crick argued that any connection between codons and amino acids is only the result of an "accident". This question is fundamental to understand the nature of specific protein-nucleic acid interactions. RESULTS: The nature of specific protein-nucleic acid interaction between restriction endonucleases (RE) and their recognition sequences (RS) was studied by bioinformatics methods. It was found that the frequency of 5-6 residue long RS-like oligonucleotides is unexpectedly high in the nucleic acid sequence of the corresponding RE (p < 0.05 and p < 0.001 respectively, n = 7). There is an extensive conservation of these RS-like sequences in RE isoschizomers. A review of the seven available crystallographic studies showed that the amino acids coded by codons that are subsets of recognition sequences were often closely located to the RS itself and they were in many cases directly adjacent to the codon-like triplets in the RS.Fifty-five examples of this codon-amino acid co-localization are found and analyzed, which represents 41.5% of total 132 amino acids which are localized within 8 A distance to the C1' atoms in the DNA. The average distance between the closest atoms in the codons and amino acids is 5.5 +/- 0.2 A (mean +/- S.E.M, n = 55), while the distance between the nitrogen and oxygen atoms of the co-localized molecules is significantly shorter, (3.4 +/- 0.2 A, p < 0.001, n = 15), when positively charged amino acids are involved. This is indicating that an interaction between the nucleic- and amino acids might occur. CONCLUSION: We interpret these results in favor of Woese and suggest that the genetic code is "rational" and there is a stereospecific relationship between the codes and the amino acids. PMID- 15113407 TI - SUPFAM: a database of sequence superfamilies of protein domains. AB - BACKGROUND: SUPFAM database is a compilation of superfamily relationships between protein domain families of either known or unknown 3-D structure. In SUPFAM, sequence families from Pfam and structural families from SCOP are associated, using profile matching, to result in sequence superfamilies of known structure. Subsequently all-against-all family profile matches are made to deduce a list of new potential superfamilies of yet unknown structure. DESCRIPTION: The current version of SUPFAM (release 1.4) corresponds to significant enhancements and major developments compared to the earlier and basic version. In the present version we have used RPS-BLAST, which is robust and sensitive, for profile matching. The reliability of connections between protein families is ensured better than before by use of benchmarked criteria involving strict e-value cut-off and a minimal alignment length condition. An e-value based indication of reliability of connections is now presented in the database. Web access to a RPS-BLAST-based tool to associate a query sequence to one of the family profiles in SUPFAM is available with the current release. In terms of the scientific content the present release of SUPFAM is entirely reorganized with the use of 6190 Pfam families and 2317 structural families derived from SCOP. Due to a steep increase in the number of sequence and structural families used in SUPFAM the details of scientific content in the present release are almost entirely complementary to previous basic version. Of the 2286 families, we could relate 245 Pfam families with apparently no structural information to families of known 3-D structures, thus resulting in the identification of new families in the existing superfamilies. Using the profiles of 3904 Pfam families of yet unknown structure, an all-against-all comparison involving sequence-profile match resulted in clustering of 96 Pfam families into 39 new potential superfamilies. CONCLUSION: SUPFAM presents many non-trivial superfamily relationships of sequence families involved in a variety of functions and hence the information content is of interest to a wide scientific community. The grouping of related proteins without a known structure in SUPFAM is useful in identifying priority targets for structural genomics initiatives and in the assignment of putative functions. Database URL: http://pauling.mbu.iisc.ernet.in/~supfam. PMID- 15113408 TI - CisOrtho: a program pipeline for genome-wide identification of transcription factor target genes using phylogenetic footprinting. AB - BACKGROUND: All known genomes code for a large number of transcription factors. It is important to develop methods that will reveal how these transcription factors act on a genome wide level, that is, through what target genes they exert their function. RESULTS: We describe here a program pipeline aimed at identifying transcription factor target genes in whole genomes. Starting from a consensus binding site, represented as a weight matrix, potential sites in a pre-filtered genome are identified and then further filtered by assessing conservation of the putative site in the genome of a related species, a process called phylogenetic footprinting. CisOrtho has been successfully used to identify targets for two homeodomain transcription factors in the genomes of the nematodes Caenorhabditis elegans and Caenorhabditis briggsae. CONCLUSIONS: CisOrtho will identify targets of other nematode transcription factors whose DNA binding specificity is known and can be easily adapted to search other genomes for transcription factor targets. PMID- 15113409 TI - Computational protein biomarker prediction: a case study for prostate cancer. AB - BACKGROUND: Recent technological advances in mass spectrometry pose challenges in computational mathematics and statistics to process the mass spectral data into predictive models with clinical and biological significance. We discuss several classification-based approaches to finding protein biomarker candidates using protein profiles obtained via mass spectrometry, and we assess their statistical significance. Our overall goal is to implicate peaks that have a high likelihood of being biologically linked to a given disease state, and thus to narrow the search for biomarker candidates. RESULTS: Thorough cross-validation studies and randomization tests are performed on a prostate cancer dataset with over 300 patients, obtained at the Eastern Virginia Medical School using SELDI-TOF mass spectrometry. We obtain average classification accuracies of 87% on a four-group classification problem using a two-stage linear SVM-based procedure and just 13 peaks, with other methods performing comparably. CONCLUSIONS: Modern feature selection and classification methods are powerful techniques for both the identification of biomarker candidates and the related problem of building predictive models from protein mass spectrometric profiles. Cross-validation and randomization are essential tools that must be performed carefully in order not to bias the results unfairly. However, only a biological validation and identification of the underlying proteins will ultimately confirm the actual value and power of any computational predictions. PMID- 15113410 TI - A web services choreography scenario for interoperating bioinformatics applications. AB - BACKGROUND: Very often genome-wide data analysis requires the interoperation of multiple databases and analytic tools. A large number of genome databases and bioinformatics applications are available through the web, but it is difficult to automate interoperation because: 1) the platforms on which the applications run are heterogeneous, 2) their web interface is not machine-friendly, 3) they use a non-standard format for data input and output, 4) they do not exploit standards to define application interface and message exchange, and 5) existing protocols for remote messaging are often not firewall-friendly. To overcome these issues, web services have emerged as a standard XML-based model for message exchange between heterogeneous applications. Web services engines have been developed to manage the configuration and execution of a web services workflow. RESULTS: To demonstrate the benefit of using web services over traditional web interfaces, we compare the two implementations of HAPI, a gene expression analysis utility developed by the University of California San Diego (UCSD) that allows visual characterization of groups or clusters of genes based on the biomedical literature. This utility takes a set of microarray spot IDs as input and outputs a hierarchy of MeSH Keywords that correlates to the input and is grouped by Medical Subject Heading (MeSH) category. While the HTML output is easy for humans to visualize, it is difficult for computer applications to interpret semantically. To facilitate the capability of machine processing, we have created a workflow of three web services that replicates the HAPI functionality. These web services use document-style messages, which means that messages are encoded in an XML-based format. We compared three approaches to the implementation of an XML-based workflow: a hard coded Java application, Collaxa BPEL Server and Taverna Workbench. The Java program functions as a web services engine and interoperates with these web services using a web services choreography language (BPEL4WS). CONCLUSION: While it is relatively straightforward to implement and publish web services, the use of web services choreography engines is still in its infancy. However, industry-wide support and push for web services standards is quickly increasing the chance of success in using web services to unify heterogeneous bioinformatics applications. Due to the immaturity of currently available web services engines, it is still most practical to implement a simple, ad-hoc XML-based workflow by hard coding the workflow as a Java application. For advanced web service users the Collaxa BPEL engine facilitates a configuration and management environment that can fully handle XML-based workflow. PMID- 15113411 TI - Biochemical Network Stochastic Simulator (BioNetS): software for stochastic modeling of biochemical networks. AB - BACKGROUND: Intrinsic fluctuations due to the stochastic nature of biochemical reactions can have large effects on the response of biochemical networks. This is particularly true for pathways that involve transcriptional regulation, where generally there are two copies of each gene and the number of messenger RNA (mRNA) molecules can be small. Therefore, there is a need for computational tools for developing and investigating stochastic models of biochemical networks. RESULTS: We have developed the software package Biochemical Network Stochastic Simulator (BioNetS) for efficiently and accurately simulating stochastic models of biochemical networks. BioNetS has a graphical user interface that allows models to be entered in a straightforward manner, and allows the user to specify the type of random variable (discrete or continuous) for each chemical species in the network. The discrete variables are simulated using an efficient implementation of the Gillespie algorithm. For the continuous random variables, BioNetS constructs and numerically solves the appropriate chemical Langevin equations. The software package has been developed to scale efficiently with network size, thereby allowing large systems to be studied. BioNetS runs as a BioSpice agent and can be downloaded from http://www.biospice.org. BioNetS also can be run as a stand alone package. All the required files are accessible from http://x.amath.unc.edu/BioNetS. CONCLUSIONS: We have developed BioNetS to be a reliable tool for studying the stochastic dynamics of large biochemical networks. Important features of BioNetS are its ability to handle hybrid models that consist of both continuous and discrete random variables and its ability to model cell growth and division. We have verified the accuracy and efficiency of the numerical methods by considering several test systems. PMID- 15113412 TI - SIGI: score-based identification of genomic islands. AB - BACKGROUND: Genomic islands can be observed in many microbial genomes. These stretches of DNA have a conspicuous composition with regard to sequence or encoded functions. Genomic islands are assumed to be frequently acquired via horizontal gene transfer. For the analysis of genome structure and the study of horizontal gene transfer, it is necessary to reliably identify and characterize these islands. RESULTS: A scoring scheme on codon frequencies Score_G1G2(cdn) = log(f_G2(cdn) / f_G1(cdn)) was utilized. To analyse genes of a species G1 and to test their relatedness to species G2, scores were determined by applying the formula to log-odds derived from mean codon frequencies of the two genomes. A non redundant set of nearly 400 codon usage tables comprising microbial species was derived; its members were used alternatively at position G2. Genes having at least one score value above a species-specific and dynamically determined cut-off value were analysed further. By means of cluster analysis, genes were identified that comprise clusters of statistically significant size. These clusters were predicted as genomic islands. Finally and individually for each of these genes, the taxonomical relation among those species responsible for significant scores was interpreted. The validity of the approach and its limitations were made plausible by an extensive analysis of natural genes and synthetic ones aimed at modelling the process of gene amelioration. CONCLUSIONS: The method reliably allows to identify genomic island and the likely origin of alien genes. PMID- 15113413 TI - Network analysis of metabolic enzyme evolution in Escherichia coli. AB - BACKGROUND: The two most common models for the evolution of metabolism are the patchwork evolution model, where enzymes are thought to diverge from broad to narrow substrate specificity, and the retrograde evolution model, according to which enzymes evolve in response to substrate depletion. Analysis of the distribution of homologous enzyme pairs in the metabolic network can shed light on the respective importance of the two models. We here investigate the evolution of the metabolism in E. coli viewed as a single network using EcoCyc. RESULTS: Sequence comparison between all enzyme pairs was performed and the minimal path length (MPL) between all enzyme pairs was determined. We find a strong over representation of homologous enzymes at MPL 1. We show that the functionally similar and functionally undetermined enzyme pairs are responsible for most of the over-representation of homologous enzyme pairs at MPL 1. CONCLUSIONS: The retrograde evolution model predicts that homologous enzymes pairs are at short metabolic distances from each other. In general agreement with previous studies we find that homologous enzymes occur close to each other in the network more often than expected by chance, which lends some support to the retrograde evolution model. However, we show that the homologous enzyme pairs which may have evolved through retrograde evolution, namely the pairs that are functionally dissimilar, show a weaker over-representation at MPL 1 than the functionally similar enzyme pairs. Our study indicates that, while the retrograde evolution model may have played a small part, the patchwork evolution model is the predominant process of metabolic enzyme evolution. PMID- 15113414 TI - Electroacupuncture enhances extragonadal aromatization in ovariectomized rats. AB - BACKGROUND: Repeated electroacupuncture (EA) stimulation is known to stimulate the activity of the hypothalamus-pituitary-adrenal axis, and to enhance the circulation level of estrogen in the ovariectomized rats. To explore the source of the increased circulation estrogen, the extragonadal aromatization was detected. METHODS: Female rats were divided into five groups: 1) intact (INT), 2) intact with EA in specific points (INT+EA), 3) ovariectomized (OVX), 4) ovariectomized with EA in specific points (OVX+EA) and 5) ovariectomized with EA in non-specific points (OVX+C). Radiometric assay, Western blot and RT-PCR were adopted to determine the extragonadal aromatization in subcutaneous abdominal (SA) adipose and liver tissues of rats. The blood concentrations of estrogen, testosterone and corticosterone were measured by radioimmunoassay. RESULTS: The aromatase activities of the SA adipose and liver tissues in the OVX+EA rats increased significantly (p < 0.01) compared with those in the INT, INT+EA and OVX rats. The 58-kDa aromatase protein and aromatase mRNA expressions normalized to beta-actin in the OVX+EA rats' SA adipose tissues showed higher levels than those from corresponding tissues in the INT and INT+EA rats (p < 0.05). And the ratios of aromatase mRNA and protein to beta-actin in the OVX+EA rats' liver tissues increased significantly compared with those in the OVX rats (p < 0.05). Furthermore, blood estrogen and corticosterone concentrations showed significant increase in the OVX+EA rats compared with the concentrations in the OVX and OVX+C rats (p < 0.05), but no statistical disparity occurred on the blood testosterone concentrations between the OVX+EA rats and the OVX ones. CONCLUSION: Both the subcutaneous abdominal adipose and the liver tissues contributed to the effects of electroacupuncture on the extragonadal aromatization to promote the blood concentrations of estrogen in the ovariectomized rats. PMID- 15113415 TI - Epigenetic CRBP downregulation appears to be an evolutionarily conserved (human and mouse) and oncogene-specific phenomenon in breast cancer. AB - BACKGROUND: The cellular retinol binding protein I gene (CRBP) is downregulated in a subset of human breast cancers and in MMTV-Myc induced mouse mammary tumors. Functional studies suggest that CRBP downregulation contributes to breast tumor progression. What is the mechanism underlying CRBP downregulation in cancer? Here we investigated the hypothesis that CRBP is epigenetically silenced through DNA hypermethylation in human and mouse breast cancer. RESULTS: Bisulfite sequencing of CRBP in a panel of 6 human breast cancer cell lines demonstrated that, as a rule, CRBP hypermethylation is closely and inversely related to CRBP expression and identified one exception to this rule. Treatment with 5-azacytidine, a DNA methyltransferase inhibitor, led to CRBP reexpression, supporting the hypothesis that CRBP hypermethylation is a proximal cause of CRBP silencing. In some cells CRBP reexpression was potentiated by co-treatment with retinoic acid, an inducer of CRBP, and trichostatin A, a histone deacetylase inhibitor. Southern blot analysis of a small panel of human breast cancer specimens identified one case characterized by extensive CRBP hypermethylation, in association with undetectable CRBP mRNA and protein. Bisulfite sequencing of CRBP in MMTV-Myc and MMTV-Neu/NT mammary tumor cell lines extended the rule of CRBP hypermethylation and silencing (both seen in MMTV-Myc but not MMTV-Neu/NT cells) from human to mouse breast cancer and suggested that CRBP hypermethylation is an oncogene specific event. CONCLUSION: CRBP hypermethylation appears to be an evolutionarily conserved and principal mechanism of CRBP silencing in breast cancer. Based on the analysis of transgenic mouse mammary tumor cells, we hypothesize that CRBP silencing in human breast cancer may be associated with a specific oncogenic signature. PMID- 15113416 TI - Metabolic and cardiovascular improvements after biliopancreatic diversion in a severely obese patient. AB - BACKGROUND: Severe obesity is associated with important morbidity and increased mortality. The successes of lifestyle modifications and drug therapy have been partial and mostly unsustained in reducing obesity and its comorbidities. Bariatric surgery, particularly biliopancreatic diversion with duodenal switch reduces efficiently excess body weight and improves metabolic and cardiovascular functions. CASE PRESENTATION: A 56-year-old man with severe clinical obesity underwent a biliopancreatic diversion with a duodenal switch after unsuccessful treatment with weight loss pharmacotherapy. He had diabetes, hypertension and sleep apnea syndrome and was on three medications for hypertension and two hypoglycemic agents in addition to > 200 insulin units daily. Eleven months after the surgery, he had lost 40% of his body weight. The lipid profile showed great improvement and the hypertension and diabetes were more easily controlled with no more insulin needed. The pseudonormalized pattern of left ventricular diastolic function improved and ventricular walls showed decreased thickness. CONCLUSION: Biliopancreatic diversion may bring metabolic and cardiovascular benefits in severely obese patients from a cardiovascular perspective. PMID- 15113417 TI - Estimation of hospital emergency room data using OTC pharmaceutical sales and least mean square filters. AB - BACKGROUND: Surveillance of Over-the-Counter pharmaceutical (OTC) sales as a potential early indicator of developing public health conditions, in particular in cases of interest to Bioterrorism, has been suggested in the literature. The data streams of interest are quite non-stationary and we address this problem from the viewpoint of linear adaptive filter theory: the clinical data is the primary channel which is to be estimated from the OTC data that form the reference channels. METHOD: The OTC data are grouped into a few categories and we estimate the clinical data using each individual category, as well as using a multichannel filter that encompasses all the OTC categories. The estimation (in the least mean square sense) is performed using an FIR (Finite Impulse Response) filter and the normalized LMS algorithm. RESULTS: We show all estimation results and present a table of effectiveness of each OTC category, as well as the effectiveness of the combined filtering operation. Individual group results clearly show the effectiveness of each particular group in estimating the clinical hospital data and serve as a guide as to which groups have sustained correlations with the clinical data. CONCLUSION: Our results indicate that Multichannel adaptive FIR least squares filtering is a viable means of estimating public health conditions from OTC sales, and provide quantitative measures of time dependent correlations between the clinical data and the OTC data channels. PMID- 15113418 TI - Characterization of digital medical images utilizing support vector machines. AB - BACKGROUND: In this paper we discuss an efficient methodology for the image analysis and characterization of digital images containing skin lesions using Support Vector Machines and present the results of a preliminary study. METHODS: The methodology is based on the support vector machines algorithm for data classification and it has been applied to the problem of the recognition of malignant melanoma versus dysplastic naevus. Border and colour based features were extracted from digital images of skin lesions acquired under reproducible conditions, using basic image processing techniques. Two alternative classification methods, the statistical discriminant analysis and the application of neural networks were also applied to the same problem and the results are compared. RESULTS: The SVM (Support Vector Machines) algorithm performed quite well achieving 94.1% correct classification, which is better than the performance of the other two classification methodologies. The method of discriminant analysis classified correctly 88% of cases (71% of Malignant Melanoma and 100% of Dysplastic Naevi), while the neural networks performed approximately the same. CONCLUSION: The use of a computer-based system, like the one described in this paper, is intended to avoid human subjectivity and to perform specific tasks according to a number of criteria. However the presence of an expert dermatologist is considered necessary for the overall visual assessment of the skin lesion and the final diagnosis. PMID- 15113419 TI - Critical reflections on evidence, ethics and effectiveness in the management of tuberculosis: public health and global perspectives. AB - BACKGROUND: Tuberculosis is a major cause of morbidity and mortality globally. Recent scholarly attention to public health ethics provides an opportunity to analyze several ethical issues raised by the global tuberculosis pandemic. DISCUSSION: Recently articulated frameworks for public health ethics emphasize the importance of effectiveness in the justification of public health action. This paper critically reviews the relationship between these frameworks and the published evidence of effectiveness of tuberculosis interventions, with a specific focus on the controversies engendered by the endorsement of programs of service delivery that emphasize direct observation of therapy. The role of global economic inequities in perpetuating the tuberculosis pandemic is also discussed. SUMMARY: Tuberculosis is a complex but well understood disease that raises important ethical challenges for emerging frameworks in public health ethics. The exact role of effectiveness as a criterion for judging the ethics of interventions needs greater discussion and analysis. Emerging frameworks are silent about the economic conditions contributing to the global burden of illness associated with tuberculosis and this requires remediation. PMID- 15113420 TI - Cluster randomisation or randomised consent as an appropriate methodology for trials in palliative care: a feasibility study [ISRCTN60243484]. AB - BACKGROUND: Although guidelines for the care of the dying patient exist the evidence base to support the guidelines is poor. Some of the factors contributing to this include failure to recruit to trials, protective healthcare professionals and subsequent attrition from trials due to the death of the patients. Recent studies report favourably on the use of cluster randomisation as an appropriate methodology for use in this patient group. METHODS/DESIGN: A feasibility study, exploring two types of randomisation as appropriate methodology for trials involving dying patients. Cluster randomisation and randomised consent will be utilised following a crossover design at two sites, one oncology ward and one Macmillan unit within the Northwest Wales NHS Trust. All patients commencing on the Integrated Care Pathway (ICP) for the Last Days of Life will be eligible for inclusion in the study. Using the hypothesis that it is not necessary to prescribe an anti-emetic medication when setting up a syringe driver for the dying patient, the study will evaluate different models of research methodology. DISCUSSION: The identification of the most appropriate methodology for use in studies concerning this patient group will inform the development of future clinical studies. Furthermore, the outcomes of this feasibility study will inform the development, of a proposal seeking funding for Wales-wide trials in palliative care. The identification of an appropriate methodology will provide a starting point for the establishment of a robust evidence base for the care of the dying patient. PMID- 15113421 TI - Crystal structure of the Habc domain of neuronal syntaxin from the squid Loligo pealei reveals conformational plasticity at its C-terminus. AB - BACKGROUND: Intracellular membrane fusion processes are mediated by the spatial and temporal control of SNARE complex assembly that results in the formation of a four-helical bundle, composed of one vesicle SNARE and three target membrane SNARE polypeptide chains. Syntaxins are essential t-SNAREs and are characterized by an N-terminal Habc domain, a flexible linker region, a coiled-coil or SNARE motif and a membrane anchor. The N-terminal Habc domain fulfills important regulatory functions while the coiled-coil motif, present in all SNAREs, is sufficient for SNARE complex formation, which is thought to drive membrane fusion. RESULTS: Here we report the crystal structure of the Habc domain of neuronal syntaxin from the squid Loligo pealei, s-syntaxin. Squid Habc crystallizes as a dimer and the monomer structure consists of a three-helical bundle. One molecule is strikingly similar to mammalian syntaxin 1A while the second one shows a structural deviation from the common fold in that the C terminal part of helix C unwinds and adopts an extended conformation. CONCLUSION: Conservation of surface residues indicates that the cytosolic part of s-syntaxin can adopt an auto-inhibitory closed conformation that may bind squid neuronal Sec1, s-Sec1, in the same manner as observed in structure of the rat nSec1/syntaxin 1A complex. Furthermore, despite the overall structural similarity, the observed changes at the C-terminus of one molecule indicate structural plasticity in neuronal syntaxin. Implications of the structural conservation and the changes are discussed with respect to potential Habc domain binding partners such as Munc13, which facilitates the transition from the closed to the open conformation. PMID- 15113422 TI - Fast identification of folded human protein domains expressed in E. coli suitable for structural analysis. AB - BACKGROUND: High-throughput protein structure analysis of individual protein domains requires analysis of large numbers of expression clones to identify suitable constructs for structure determination. For this purpose, methods need to be implemented for fast and reliable screening of the expressed proteins as early as possible in the overall process from cloning to structure determination. RESULTS: 88 different E. coli expression constructs for 17 human protein domains were analysed using high-throughput cloning, purification and folding analysis to obtain candidates suitable for structural analysis. After 96 deep-well microplate expression and automated protein purification, protein domains were directly analysed using 1D 1H-NMR spectroscopy. In addition, analytical hydrophobic interaction chromatography (HIC) was used to detect natively folded protein. With these two analytical methods, six constructs (representing two domains) were quickly identified as being well folded and suitable for structural analysis. CONCLUSION: The described approach facilitates high-throughput structural analysis. Clones expressing natively folded proteins suitable for NMR structure determination were quickly identified upon small scale expression screening using 1D 1H-NMR and/or analytical HIC. This procedure is especially effective as a fast and inexpensive screen for the 'low hanging fruits' in structural genomics. PMID- 15113423 TI - COMe: the ontology of bioinorganic proteins. AB - BACKGROUND: Many characterised proteins contain metal ions, small organic molecules or modified residues. In contrast, the huge amount of data generated by genome projects consists exclusively of sequences with almost no annotation. One of the goals of the structural genomics initiative is to provide representative three-dimensional (3-D) structures for as many protein/domain folds as possible to allow successful homology modelling. However, important functional features such as metal co-ordination or a type of prosthetic group are not always conserved in homologous proteins. So far, the problem of correct annotation of bioinorganic proteins has been largely ignored by the bioinformatics community and information on bioinorganic centres obtained by methods other than crystallography or NMR is only available in literature databases. RESULTS: COMe (Co-Ordination of Metals) represents the ontology for bioinorganic and other small molecule centres in complex proteins. COMe consists of three types of entities: 'bioinorganic motif' (BIM), 'molecule' (MOL), and 'complex proteins' (PRX), with each entity being assigned a unique identifier. A BIM consists of at least one centre (metal atom, inorganic cluster, organic molecule) and two or more endogenous and/or exogenous ligands. BIMs are represented as one-dimensional (1-D) strings and 2-D diagrams. A MOL entity represents a 'small molecule' which, when in complex with one or more polypeptides, forms a functional protein. The PRX entities refer to the functional proteins as well as to separate protein domains and subunits. The complex proteins in COMe are subdivided into three categories: (i) metalloproteins, (ii) organic prosthetic group proteins and (iii) modified amino acid proteins. The data are currently stored in both XML format and a relational database and are available at http://www.ebi.ac.uk/come/. CONCLUSION: COMe provides the classification of proteins according to their 'bioinorganic' features and thus is orthogonal to other classification schemes, such as those based on sequence similarity, 3-D fold, enzyme activity, or biological process. The hierarchical organisation of the controlled vocabulary allows both for annotation and querying at different levels of granularity. PMID- 15113424 TI - Human cortical perfusion and the arterial pulse: a near-infrared spectroscopy study. AB - BACKGROUND: The pulsatile nature of the arterial pulse induces a pulsatile perfusion pattern which can be observed in human cerebral cortex with non invasive near-infrared spectroscopy. The present study attempts to establish a quantitative relation between these two events, even in situations of very weak signal-to-noise ratio in the cortical perfusion signal. The arterial pulse pattern was extracted from the left middle finger by means of plethesmographic techniques. Changes in cortical perfusion were detected with a continuous-wave reflectance spectrophotometer on the scalp overlying the left prefrontal cortex. Cross-correlation analysis was performed to provide evidence for a causal relation between the arterial pulse and relative changes in cortical total hemoglobin. In addition, the determination of the statistical significance of this relation was established by the use of phase-randomized surrogates. RESULTS: The results showed statistically significant cross correlation between the arterial and perfusion signals. CONCLUSIONS: The approach designed in the present study can be utilized for a quantitative and continuous assessment of the perfusion states of the cerebral cortex in experimental and clinical settings even in situations of extremely low signal-to-noise ratio. PMID- 15113425 TI - Heart rate variability and short duration spaceflight: relationship to post flight orthostatic intolerance. AB - BACKGROUND: Upon return from space many astronauts experience symptoms of orthostatic intolerance. Research has implicated altered autonomic cardiovascular regulation due to spaceflight with further evidence to suggest that there might be pre-flight autonomic indicators of post-flight orthostatic intolerance. We used heart rate variability (HRV) to determine whether autonomic regulation of the heart in astronauts who did or did not experience post-flight orthostatic intolerance was different pre-flight and/or was differentially affected by short duration (8-16 days) spaceflight. HRV data from ten-minute stand tests collected from the 29 astronauts 10 days pre-flight, on landing day and three days post flight were analysed using coarse graining spectral analysis. From the total power (PTOT), the harmonic component was extracted and divided into high (PHI: >0.15 Hz) and low (PLO: = 0.15 Hz) frequency power regions. Given the distribution of autonomic nervous system activity with frequency at the sinus node, PHI/PTOT was used as an indicator of parasympathetic activity; PLO/PTOT as an indicator of sympathetic activity; and, PLO/PHI as an estimate of sympathovagal balance. RESULTS: Twenty-one astronauts were classified as finishers, and eight as non-finishers, based on their ability to remain standing for 10 minutes on landing day. Pre-flight, non-finishers had a higher supine PHI/PTOT than finishers. Supine PHI/PTOT was the same pre-flight and on landing day in the finishers; whereas, in the non-finishers it was reduced. The ratio PLO/PHI was lower in non-finishers compared to finishers and was unaffected by spaceflight. Pre-flight, both finishers and non-finishers had similar supine values of PLO/PTOT, which increased from supine to stand. Following spaceflight, only the finishers had an increase in PLO/PTOT from supine to stand. CONCLUSIONS: Both finishers and non-finishers had an increase in sympathetic activity with stand on pre-flight, yet only finishers retained this response on landing day. Non-finishers also had lower sympathovagal balance and higher pre-flight supine parasympathetic activity than finishers. These results suggest pre-flight autonomic status and post-flight impairment in autonomic control of the heart may contribute to orthostatic intolerance. The mechanism by which higher pre-flight parasympathetic activity might contribute to post-flight orthostatic intolerance is not understood and requires further investigation. PMID- 15113426 TI - Detecting and minimizing zinc contamination in physiological solutions. AB - BACKGROUND: To explore the role of zinc (Zn) in cellular physiology it is important to be able to control and quantify the level of Zn contamination in experimental solutions. A technique that relies on a Zn-sensitive fluorimetric probe is introduced for measuring Zn concentrations as low as 100 pM. The method depends on the combination of the Zn-probe FluoZin-3 together with a slow Zn chelator, Ca-EDTA, that reduces the background Zn levels and allows repeated measurements in the same solution. RESULTS: The method was used to determine which common labware items could leach Zn into solution. Contamination was predictably found to arise from stainless steel and glass. Perhaps less expectedly it was also introduced by methacrylate cuvettes, plastic tissue culture dishes and other plastic labware. The release of nickel from stainless steel electrodes was also imaged using the fluorescent probe Newport Green. CONCLUSION: Zn contamination may arise from rather unexpected sources; it is important that all aspects and components used in the course of an experiment be analyzed for the possibility of introducing contaminants. PMID- 15113427 TI - The defH9-iaaM auxin-synthesizing gene increases plant fecundity and fruit production in strawberry and raspberry. AB - BACKGROUND: The DefH9-iaaM gene fusion which is expressed specifically in placenta/ovules and promotes auxin-synthesis confers parthenocarpic fruit development to eggplant, tomato and tobacco. Transgenic DefH9-iaaM eggplants and tomatoes show increased fruit production due mainly to an improved fruit set. However, the weight of the fruits is also frequently increased. RESULTS: DefH9 iaaM strawberry and raspberry plants grown under standard cultivation conditions show a significant increase in fruit number and size and fruit yield. In all three Rosaceae species tested, Fragaria vesca, Fragaria x ananassa and Rubus idaeus, DefH9-iaaM plants have an increased number of flowers per inflorescence and an increased number of inflorescences per plant. This results in an increased number of fruits per plant. Moreover, the weight and size of transgenic fruits was also increased. The increase in fruit yield was approximately 180% in cultivated strawberry, 140% in wild strawberry, and 100% in raspberry. The DefH9 iaaM gene is expressed in the flower buds of all three species. The total IAA (auxin) content of young flower buds of strawberry and raspberry expressing the DefH9-iaaM gene is increased in comparison to untransformed flower buds. The DefH9-iaaM gene promotes parthenocarpy in emasculated flowers of both strawberry and raspberry. CONCLUSIONS: The DefH9-iaaM gene is expressed and biologically active in Rosaceae. The DefH9-iaaM gene can be used, under cultivation conditions that allow pollination and fertilization, to increase fruit productivity significantly in Rosaceae species. The finding that the DefH9-iaaM auxin synthesizing gene increases the number of inflorescences per plant and the number of flowers per inflorescence indicates that auxin plays a role in plant fecundity in these three perennial Rosaceae species. PMID- 15113428 TI - The limits of log-ratios. AB - BACKGROUND: DNA microarray assays typically compare two biological samples and present the results of those comparisons gene-by-gene as the logarithm base two of the ratio of the measured expression levels for the two samples. RESULTS: Because of the fixed dynamic range of fluorescence and other detection systems, there is a limit to the range of comparisons that can be made using any array technology, and this must be taken into account when interpreting the results of any such analysis. CONCLUSIONS: The dynamic range of microarray data collection systems results in limits in the comparative analyses that can be derived from such measurements and suggests that optimal results can be obtained by making measurements that avoid the boundaries of that dynamic range. PMID- 15113429 TI - 'Furuncular myiasis of the breast caused by the larvae of the Tumbu fly (Cordylobia anthropophaga)'. AB - BACKGROUND: Cutaneous myiasis of the breast due to infestation by the larva of Cordylobia anthropophaga is rare. To the best of our knowledge, only one case has been reported in the English literature. This rarity calls for an awareness of its possibility as a cause of furuncular breast lesions, especially in areas where the C. anthropophaga (Tumbu fly) is endemic or in patients returning from such areas. As it can be easily confused with other furuncular breast lesions (like tuberculosis, mycosis, actinomycosis, furunculosis, chronic breast abscess and fungating malignancies), this awareness is important to avoid misdiagnosis or delay in diagnosis. We present a case of furuncular breast myiasis due to the larvae of C. anthropophaga earlier misdiagnosed as mastitis in a patient living in tropical Africa (Nigeria) where the Tumbu fly is endemic. CASE PRESENTATION: We report a 70 year old woman who presented with a week history of itchy multiple discharging sinuses of the right breast. The sinuses contained wriggling larvae of C. anthropophaga. Fourteen larvae were extracted from the breast and the sinuses healed quite well after the extraction. CONCLUSIONS: Cutaneous myiasis of the breast is rare, hence, an awareness of its clinical features is necessary when a patient presents with furuncular skin lesions especially in endemic areas or people returning from such areas. Diagnosis is mainly clinical and lesions heal well after the extraction of the larvae. Preventive measures such as ironing after drying of dresses and a good personal hygiene are crucial in controlling C. anthropophaga infestation. PMID- 15113430 TI - "Primary" aggressive chondroblastoma of the humerus: a case report. AB - BACKGROUND: Chondroblastomas are rare epiphyseal bone tumors. Very few cases with extra-cortical aggressive soft tissue invasion or metastasis are reported. CASE PRESENTATION: We report a 28 year-old adult male who presented with a large swelling over the left shoulder region. Pre-operative imaging revealed a large tumor arising from upper end of humerus with extensive soft tissue involvement necessitating a fore-quarter amputation. Patient received adjuvant radiation. CONCLUSIONS: This patient is one of the largest chondroblastomas to be reported. Although chondroblastomas are typically benign, rarely they can be locally aggressive or metastatic. Early diagnosis and institution of proper primary therapy would prevent mutilating surgeries and recurrences. PMID- 15113431 TI - Multiple parallel skin markers for minimal incision lumbar disc surgery; a technical note. AB - BACKGROUND: Spinal surgery depends on accurate localization to prevent incorrect surgical approaches. The trend towards minimally invasive surgery that minimizes surgical exposure and reduces postoperative pain increasingly requires surgeons to accurately determine the operative level before an incision is made. Preoperative localization with a C-arm image intensifier is popular, but the exposure of both patients and theatre staff to radiation is a disadvantage, as well as being time-consuming. METHODS: We describe a simple surgical tool developed to help localize exact spinal levels in conjunction with a simple AP X ray film immediately before surgery. Multiple parallel skin markers were made using a circular oven rack comprising multiple 1.5 cm spaced parallel wires attached to a circular outside rim. The longest line was placed on the line of the postero-superior iliac spine (PSIS) over the junction of the L5-S1 region. RESULTS AND CONCLUSIONS: Based on the film taken, the incision can be accurately made at the intended level. The incision wound can be minimized to 3.0 cm even when using conventional disc surgery instruments. PMID- 15113433 TI - Serum interleukin-5 levels are elevated in mild and moderate persistent asthma irrespective of regular inhaled glucocorticoid therapy. AB - BACKGROUND: Interleukin-5 (IL-5) is thought to play a pivotal role in the pathogenesis of asthma. High levels of circulating IL-5 have been documented in acute asthma. However, serum IL-5 levels in mild to moderate asthmatics and the influence of regular use of inhaled glucocorticoids, is not known. METHODS: Fifty six asthmatics and 56 age and sex matched controls were recruited prospectively from an outpatient department. Information on asthma severity and treatment was gathered by a questionnaire. Serum IL-5, total IgE and specific IgE levels were measured in a blinded fashion. RESULTS: There were 32 atopic and 24 non-atopic mild-to-moderate asthmatics. The median serum IL-5 levels in atopic asthmatics (9.5 pg/ml) and in non-atopic asthmatics (8.1 pg/ml) were significantly higher than in normal controls (4.4 pg/ml, both p < 0.003). However, median serum IL-5 levels in atopic and non-atopic asthmatics were not significantly different. The median serum IL-5 level was insignificantly higher in fourteen moderate persistent asthmatics (10.6 pg/ml) compared to forty-two mild persistent asthmatics (7.3 pg/ml) (p = 0.13). The median serum IL-5 levels in asthmatics using regular inhaled steroids (7.8 pg/ml) was not significantly different from those not using inhaled steroids (10.2 pg/ml). Furthermore, serum total IgE levels and eosinophil counts were not significantly different in those using versus those not using inhaled glucocorticoids. CONCLUSION: Serum IL-5 levels are elevated in mild and moderate persistent atopic and non-atopic asthmatics. Regular use of inhaled glucocorticoids may not abrogate the systemic Th2 type of inflammatory response in mild-moderate persistent asthma. PMID- 15113432 TI - An experimental animal model of aseptic loosening of hip prostheses in sheep to study early biochemical changes at the interface membrane. AB - BACKGROUND: Aseptic loosening of hip prosthesis as it occurs in clinical cases in human patients was attributed to wear particles of the implants, the response of the tissue dominated by macrophages and the production of inflammatory mediators and matrix degrading enzymes; however, the cascade of events initiating the process and their interaction regarding the time course is still open and discussed controversially. Therefore, the goal of this study was to establish an experimental animal model in sheep allowing to follow the cascade of early mechanical and biochemical events within the interface membrane and study the sequence of how they contribute to the pathological bone resorption necessary for aseptic loosening of the implant. METHODS: A cemented modular system (Biomedtrix) was used as a hip replacement in 24 adult Swiss Alpine sheep, with one group receiving a complete cement mantle as controls (n = 12), and the other group a cement mantle with a standardized, lateral, primary defect in the cement mantle (n = 12). Animals were followed over time for 2 and 8.5 months (n = 6 each). After sacrifice, samples from the interface membranes were harvested from five different regions of the femur and joint capsule. Explant cell cultures were performed and supernatant of cultures were tested and assayed for nitric oxide, prostaglandin E2, caseinolytic and collagenolytic activity. RNA extraction and quantification were performed for inducible nitric oxide synthase, cyclooxygenase 2, interleukin 1, and interleukin 6. Overall differences between groups and time periods and interactions thereof were calculated using a factorial analysis of variance (ANOVA). RESULTS: The development of an interface membrane was noticed in both groups at both time points. However, in the controls the interface membrane regressed in thickness and biological activity, while both variables increased in the experimental group with the primary cement mantle defect over time. Nitric oxide (NO) and PGE2 concentrations were higher in the 8.5 months group (P < 0.0001) compared to the 2 months group with a tendency for the unstable group to have higher concentrations. The same was true for collagenolytic activity (P = 0.05), but not for caseinolytic activity that decreased over time (P < 0.0001). CONCLUSION: In this study, a primary cement mantle defect of the femoral shaft elicited biomechanical instability and biochemical changes over time in an experimental animal study in sheep, that resembled the changes described at the bone cement-interface in aseptic loosening of total hip prosthesis in humans. The early biochemical changes may well explain the pathologic bone resorption and formation of an interface membrane as is observed in clinical cases. This animal model may aid in future studies aiming at prevention of aseptic loosening of hip prosthesis and reflect some aspects of the pathogenesis involved. PMID- 15113434 TI - Measuring the effects of acupuncture and homoeopathy in general practice: an uncontrolled prospective documentation approach. AB - BACKGROUND: Despite the increasing demand for acupuncture and homoeopathy in Germany, little is known about the effects of these treatments in routine care. We set up a pragmatic documentation study in general practice funded within the scope of project launched by a German health insurer. Patients were followed-up for up to four years. METHODS: The aim of the project was to study the effects and benefits of acupuncture and/or homoeopathy, and to assess patient satisfaction within a prospective documentation of over 5000 acupuncture and over 900 homoeopathy patients. As data sources, we used the documentation made available by therapists on every individual visit and a standardised quality-of life questionnaire (MOS SF-36); these were complemented by questions concerning the patient's medical history and by questions on patient satisfaction. The health insurer provided us with data on work absenteeism. RESULTS: Descriptive analyses of the main outcomes showed benefit of treatment with middle to large sized effects for the quality of life questionnaire SF-36 and about 1 point improvement on a rating scale of effects, given by doctors. Data on the treatment and the patients' and physicians' background suggests chronically ill patients treated by fairly regular schemes. CONCLUSION: Since the results showed evidence of a subjective benefit for patients from acupuncture and homoeopathy, this may account for the increase in demand for these treatments especially when patients are chronically ill and unsatisfied with the conventional treatment given previously. PMID- 15113435 TI - Validation of the diagnosis of autism in general practitioner records. AB - BACKGROUND: We report on the validity of the computerized diagnoses of autism in a large case-control study investigating the possible association between autism and the measles, mumps and rubella vaccine in the UK using the General Practitioner Research Database (GPRD). We examined anonymized copies of all relevant available clinical reports, including general practitioners' (GP) notes, consultant, speech therapy and educational psychologists reports, on 318 subjects born between 1973 and 1997 with a diagnosis of autism or a related disorder recorded in their electronic general practice record. METHODS: Data were abstracted to a case validation form allowing for the identification of developmental symptoms relevant to the diagnosis of pervasive developmental disorders (PDDs). Information on other background clinical and familial features was also abstracted. A subset of 50 notes was coded independently by 2 raters to derive reliability estimates for key clinical characteristics. RESULTS: For 294 subjects (92.5%) the diagnosis of PDD was confirmed after review of the records. Of these, 180 subjects (61.2%) fulfilled criteria for autistic disorder. The mean age at first recording of a PDD diagnosis in the GPRD database was 6.3 years (SD = 4.6). Consistent with previous estimates, the proportion of subjects experiencing regression in the course of their development was 19%. Inter-rater reliability for the presence of a PDD diagnosis was good (kappa =.73), and agreement on clinical features such as regression, age of parental recognition of first symptoms, language delay and presence of epilepsy was also good (kappas ranging from.56 to 1.0). CONCLUSIONS: This study provides evidence that the positive predictive value of a diagnosis of autism recorded in the GPRD is high. PMID- 15113436 TI - Hepatitis B virus infection in Haemodialysis Centres from Santa Catarina State, Southern Brazil. Predictive risk factors for infection and molecular epidemiology. AB - BACKGROUND: Patients under haemodialysis are considered at high risk to acquire hepatitis B virus (HBV) infection. Since few data are reported from Brazil, our aim was to assess the frequency and risk factors for HBV infection in haemodialysis patients from 22 Dialysis Centres from Santa Catarina State, south of Brazil. METHODS: This study includes 813 patients, 149 haemodialysis workers and 772 healthy controls matched by sex and age. Serum samples were assayed for HBV markers and viraemia was detected by nested PCR. HBV was genotyped by partial S gene sequencing. Univariate and multivariate statistical analyses with stepwise logistic regression analysis were carried out to analyse the relationship between HBV infection and the characteristics of patients and their Dialysis Units. RESULTS: Frequency of HBV infection was 10.0%, 2.7% and 2.7% among patients, haemodialysis workers and controls, respectively. Amidst patients, the most frequent HBV genotypes were A (30.6%), D (57.1%) and F (12.2%). Univariate analysis showed association between HBV infection and total time in haemodialysis, type of dialysis equipment, hygiene and sterilization of equipment, number of times reusing the dialysis lines and filters, number of patients per care-worker and current HCV infection. The logistic regression model showed that total time in haemodialysis, number of times of reusing the dialysis lines and filters, and number of patients per worker were significantly related to HBV infection. CONCLUSIONS: Frequency of HBV infection among haemodialysis patients at Santa Catarina state is very high. The most frequent HBV genotypes were A, D and F. The risk for a patient to become HBV positive increase 1.47 times each month of haemodialysis; 1.96 times if the dialysis unit reuses the lines and filters > or = 10 times compared with haemodialysis units which reuse < 10 times; 3.42 times if the number of patients per worker is more than five. Sequence similarity among the HBV S gene from isolates of different patients pointed out to nosocomial transmission. PMID- 15113437 TI - Attrition and bias in the MRC cognitive function and ageing study: an epidemiological investigation. AB - BACKGROUND: Any hypothesis in longitudinal studies may be affected by attrition and poor response rates. The MRC Cognitive Function and Ageing study (MRC CFAS) is a population based longitudinal study in five centres with identical methodology in England and Wales each recruiting approximately 2,500 individuals. This paper aims to identify potential biases in the two-year follow-up interviews. METHODS: Initial non-response: Those not in the baseline interviews were compared in terms of mortality to those who were in the baseline interviews at the time of the second wave interviews (1993-1996). Longitudinal attrition: Logistic regression analysis was used to examine baseline differences between individuals who took part in the two-year longitudinal wave compared with those who did not. RESULTS: Initial non-response: Individuals who moved away after sampling but before baseline interview were 1.8 times more likely to die by two years (95% Confidence interval(CI) 1.3-2.4) compared to respondents, after adjusting for age. The refusers had a slightly higher, but similar mortality pattern to responders (Odds ratio 1.2, 95%CI 1.1-1.4). Longitudinal attrition: Predictors for drop out due to death were being older, male, having impaired activities of daily living, poor self-perceived health, poor cognitive ability and smoking. Similarly individuals who refused were more likely to have poor cognitive ability, but had less years of full-time education and were more often living in their own home though less likely to be living alone. There was a higher refusal rate in the rural centres. Individuals who moved away or were uncontactable were more likely to be single, smokers, demented or depressed and were less likely to have moved if in warden-controlled accommodation at baseline. CONCLUSIONS: Longitudinal estimation of factors mentioned above could be biased, particularly cognitive ability and estimates of movements from own home to residential homes. However, these differences could also affect other investigations, particularly the estimates of incidence and longitudinal effects of health and psychiatric diseases, where the factors shown here to be associated with attrition are risk factors for the diseases. All longitudinal studies should investigate attrition and this may help with aspects of design and with the analysis of specific hypotheses. PMID- 15113439 TI - Posttraumatic Stress Disorder in patients with traumatic brain injury. AB - BACKGROUND: Severe traumatic stressors such as war, rape, or life-threatening accidents can result in a debilitating psychopathological development conceptualised as Posttraumatic Stress Disorder (PTSD). Pathological memory formation during an alarm response may set the precondition for PTSD to occur. If true, a lack of memory formation by extended unconsciousness in the course of the traumatic experience should preclude PTSD. METHODS: 46 patients from a neurological rehabilitation clinic were examined by means of questionnaires and structured clinical interviews. All patients had suffered a TBI due to an accident, but varied with respect to falling unconscious during the traumatic event. RESULTS: 27% of the sub-sample who were not unconscious for an extended period but only 3% (1 of 31 patients) who were unconscious for more than 12 hours as a result of the accident were diagnosed as having current PTSD (P <.02). Furthermore, intrusive memories proved to be far more frequent in patients who had not been unconscious. This was also the case for other re-experiencing symptoms and for psychological distress and physiological reactivity to reminders of the traumatic event. CONCLUSION: TBI and PTSD are not mutually exclusive. However, victims of accidents are unlikely to develop a PTSD if the impact to the head had resulted in an extended period of unconsciousness. PMID- 15113438 TI - Thyroid function in clinical subtypes of major depression: an exploratory study. AB - BACKGROUND: Unipolar depression might be characterized by a 'low-thyroid function syndrome'. To our knowledge, this is the first study which explores the possible relationship of DSM-IV depressive subtypes and the medium term outcome, with thyroid function. METHODS: MATERIAL: Thirty major depressive patients (DSM-IV) aged 21-60 years and 60 control subjects were included. Clinical Diagnosis: The SCAN v 2.0 and the IPDE were used. The psychometric Assessment included HDRS the HAS and the GAF scales. Free-T3, Free-T4, TSH, Thyroid Binding Inhibitory Immunoglobulins (TBII), Thyroglobulin antibodies (TA) and Thyroid Microsomal Antibodies (TMA) were measured in the serum. The Statistical analysis included 1 and 2-way MANCOVA, discriminant function analysis and Pearson Product Moment Correlation Coefficient. RESULTS: All depressive subtypes had significantly higher TBII levels in comparison to controls. Atypical patients had significantly higher TMA in comparison to controls. No significant correlation was observed between the HDRS, HAS and GAF scales and thyroid indices. Discriminant function analysis produced functions based on thyroid indices, which could moderately discriminate between diagnostic groups, but could predict good response to treatment with 89.47% chance of success. CONCLUSION: Although overt thyroid dysfunction is not common in depression, there is evidence suggesting the presence of an autoimmune process affecting the thyroid gland in depressive patients PMID- 15113440 TI - Children grow and horses race: is the adiposity rebound a critical period for later obesity? AB - BACKGROUND: The adiposity rebound is the second rise in body mass index that occurs between 3 and 7 years. An early age at adiposity rebound is known to be a risk factor for later obesity. The aim here is to clarify the connection between the age at rebound and the corresponding pattern of body mass index change, in centile terms, so as to better understand its ability to predict later fatness. DISCUSSION: Longitudinal changes in body mass index during adiposity rebound, measured both in original (kg/m2) and standard deviation (SD) score units, are studied in five hypothetical subjects. Two aspects of the body mass index curve, the body mass index centile and the rate of body mass index centile crossing, determine a child's age at rebound. A high centile and upward centile crossing are both associated separately with an early rebound, while a low centile and/or downward centile crossing correspond to a late rebound. Early adiposity rebound is a risk factor for later fatness because it identifies children whose body mass index centile is high and/or crossing upwards. Such children are likely to have a raised body mass index later in childhood and adulthood. This is an example of Peto's "horse racing effect". The association of centile crossing with later obesity is statistical not physiological, and it applies at all ages not just at rebound, so adiposity rebound cannot be considered a critical period for future obesity. Body mass index centile crossing is a more direct indicator of the underlying drive to fatness. SUMMARY: An early age at adiposity rebound predicts later fatness because it identifies children whose body mass index centile is high and/or crossing upwards. Such children are likely to have a raised body mass index later. Body mass index centile crossing is more direct than the timing of adiposity rebound for predicting later fatness. PMID- 15113441 TI - Kin-cohort estimates for familial breast cancer risk in relation to variants in DNA base excision repair, BRCA1 interacting and growth factor genes. AB - BACKGROUND: Subtle functional deficiencies in highly conserved DNA repair or growth regulatory processes resulting from polymorphic variation may increase genetic susceptibility to breast cancer. Polymorphisms in DNA repair genes can impact protein function leading to genomic instability facilitated by growth stimulation and increased cancer risk. Thus, 19 single nucleotide polymorphisms (SNPs) in eight genes involved in base excision repair (XRCC1, APEX, POLD1), BRCA1 protein interaction (BRIP1, ZNF350, BRCA2), and growth regulation (TGFss1, IGFBP3) were evaluated. METHODS: Genomic DNA samples were used in Taqman 5' nuclease assays for most SNPs. Breast cancer risk to ages 50 and 70 were estimated using the kin-cohort method in which genotypes of relatives are inferred based on the known genotype of the index subject and Mendelian inheritance patterns. Family cancer history data was collected from a series of genotyped breast cancer cases (N = 748) identified within a cohort of female US radiologic technologists. Among 2,430 female first-degree relatives of cases, 190 breast cancers were reported. RESULTS: Genotypes associated with increased risk were: XRCC1 R194W (WW and RW vs. RR, cumulative risk up to age 70, risk ratio (RR) = 2.3; 95% CI 1.3-3.8); XRCC1 R399Q (QQ vs. RR, cumulative risk up to age 70, RR = 1.9; 1.1-3.9); and BRIP1 (or BACH1) P919S (SS vs. PP, cumulative risk up to age 50, RR = 6.9; 1.6-29.3). The risk for those heterozygous for BRCA2 N372H and APEX D148E were significantly lower than risks for homozygotes of either allele, and these were the only two results that remained significant after adjusting for multiple comparisons. No associations with breast cancer were observed for: APEX Q51H; XRCC1 R280H; IGFPB3 -202A>C; TGFss1 L10P, P25R, and T263I; BRCA2 N289H and T1915M; BRIP1 -64A>C; and ZNF350 (or ZBRK1) 1845C>T, L66P, R501S, and S472P. CONCLUSION: Some variants in genes within the base-excision repair pathway (XRCC1) and BRCA1 interacting proteins (BRIP1) may play a role as low penetrance breast cancer risk alleles. Previous association studies of breast cancer and BRCA2 N372H and functional observations for APEX D148E ran counter to our findings of decreased risks. Due to the many comparisons, cautious interpretation and replication of these relationships are warranted. PMID- 15113442 TI - Computer simulation in conjunction with medical thermography as an adjunct tool for early detection of breast cancer. AB - BACKGROUND: Mathematical modelling and analysis is now accepted in the engineering design on par with experimental approaches. Computer simulations enable one to perform several 'what-if' analyses cost effectively. High speed computers and low cost of memory has helped in simulating large-scale models in a relatively shorter time frame. The possibility of extending numerical modelling in the area of breast cancer detection in conjunction with medical thermography is considered in this work. METHODS: Thermography enables one to see the temperature pattern and look for abnormality. In a thermogram there is no radiation risk as it only captures the infrared radiation from the skin and is totally painless. But, a thermogram is only a test of physiology, whereas a mammogram is a test of anatomy. It is hoped that a thermogram along with numerical modelling will serve as an adjunct tool. Presently mammogram is the 'gold-standard' in breast cancer detection. But the interpretation of a mammogram is largely dependent on the radiologist. Therefore, a thermogram that looks into the physiological changes in combination with numerical simulation performing 'what-if' analysis could act as an adjunct tool to mammography. RESULTS: The proposed framework suggested that it could reduce the occurrence of false negative/positive cases. CONCLUSION: A numerical bioheat model of a female breast is developed and simulated. The results are compared with experimental results. The possibility of this method as an early detection tool is discussed. PMID- 15113443 TI - Squamous cell carcinoma developing in the scar of Fournier's gangrene--case report. AB - BACKGROUND: Squamous cell carcinoma of the scrotum is rare and its development in the scar of Fournier's gangrene is still rarer. CASE PRESENTATION: A 65-year-old gentleman presented with a small non-healing ulcer developing on right hemi scrotum two years after the treatment for Fournier's gangrene. On histological examination it was found to be squamous cell carcinoma. He was successfully managed by surgery in the form of wide local excision and ilio-inguinal lymph node dissection followed by adjuvant radiotherapy and chemotherapy. CONCLUSIONS: Squamous cell carcinoma can develop in the scar of Fournier's gangrene after a long delay, which differentiates it from other scar carcinomas or Marjolin's ulcer. PMID- 15113444 TI - Tumor classification: molecular analysis meets Aristotle. AB - BACKGROUND: Traditionally, tumors have been classified by their morphologic appearances. Unfortunately, tumors with similar histologic features often follow different clinical courses or respond differently to chemotherapy. Limitations in the clinical utility of morphology-based tumor classifications have prompted a search for a new tumor classification based on molecular analysis. Gene expression array data and proteomic data from tumor samples will provide complex data that is unobtainable from morphologic examination alone. The growing question facing cancer researchers is, "How can we successfully integrate the molecular, morphologic and clinical characteristics of human cancer to produce a helpful tumor classification?" DISCUSSION: Current efforts to classify cancers based on molecular features ignore lessons learned from millennia of experience in biological classification. A tumor classification must include every type of tumor and must provide a unique place for each tumor within the classification. Groups within a classification inherit the properties of their ancestors and impart properties to their descendants. A classification was prepared grouping tumors according to their histogenetic development. The classification is simple (reducing the complexity of information received from the molecular analysis of tumors), comprehensive (providing a place for every tumor of man), and consistent with recent attempts to characterize tumors by cytogenetic and molecular features. The clinical and research value of this historical approach to tumor classification is discussed. SUMMARY: This manuscript reviews tumor classification and provides a new and comprehensive classification for neoplasia that preserves traditional nomenclature while incorporating information derived from the molecular analysis of tumors. The classification is provided as an open access XML document that can be used by cancer researchers to relate tumor classes with heterogeneous experimental and clinical tumor databases. PMID- 15113445 TI - The Magpie Trial follow up study: outcome after discharge from hospital for women and children recruited to a trial comparing magnesium sulphate with placebo for pre-eclampsia [ISRCTN86938761]. AB - BACKGROUND: The Magpie Trial compared magnesium sulphate with placebo for women with pre-eclampsia. 10,141 women were recruited, 8804 before delivery. Overall, 9024 children were included in the analysis of outcome at discharge from hospital. Magnesium sulphate more than halved the risk of eclampsia, and probably reduced the risk of maternal death. There did not appear to be any substantive harmful effects on the baby, in the short term. It is now important to assess whether these benefits persist, and to provide adequate reassurance about longer term safety.The main objective of the Magpie Trial Follow Up Study is to assess whether in utero exposure to magnesium sulphate has a clinically important effect on the child's chance of surviving without major neurosensory disability. Other objectives are to assess long term outcome for the mother, and to develop and assess appropriate strategies for following up large numbers of children in perinatal trials. STUDY DESIGN: Follow up is only feasible in selected centres. We therefore anticipate contacting 2800-3350 families, for 2435-2915 of whom the woman was randomised before delivery. A further 280-335 children would have been eligible for follow up if they had survived. The total sample size for the children is therefore 3080-3685, 2680-3210 of whom will have been born to women randomised before delivery.Families eligible for the follow up will be contacted, and surviving children screened using the Ages and Stages Questionnaires. Children who screen positive, and a sample of those who screen negative, will whenever possible have a paediatric and neurodevelopmental assessment. When women are contacted to ask how their child is, they will also be asked about their own health. The primary outcome is a composite measure of death or neurosensory disability for the child at 18 months. DISCUSSION: The Follow Up Study began in 2002, and now involves collaborators in 19 countries. Data collection will close at the end of 2003. PMID- 15113446 TI - Mobility and maternal position during childbirth in Tanzania: an exploratory study at four government hospitals. AB - BACKGROUND: Emerging research evidence suggests a potential benefit in being upright in the first stage of labour and a systematic review of trials suggests both benefits and harmful effects associated with being upright in the second stage of labour. Implementing evidence-based obstetric care in African countries with scarce resources is particularly challenging, and requires an understanding of the cumulative nature of science and commitment to applying the most up to date evidence to clinical decisions. In this study, we documented current practice rates, explored the barriers and opportunities to implementing these procedures from the provider perspective, and documented women's preferences and satisfaction with care. METHODS: This was an exploratory study using quantitative and qualitative methods. Practice rates were determined by exit interviews with a consecutive sample of postnatal women. Provider views were explored using semi structured interviews (with doctors and traditional birth attendants) and focus group discussions (with midwives). The study was conducted at four government hospitals, two in Dar es Salaam and two in the neighbouring Coast region, Tanzania. MAIN OUTCOME MEASURES: Practice rates for mobility during labour and delivery position; women's experiences, preferences and views about the care provided; and provider views of current practice and barriers and opportunities to evidence-based obstetric practice. RESULTS: Across all study sites more women were mobile at home (15.0%) than in the labour ward (2.9%), but movement was quite restricted at home before women were admitted to labour ward (51.6% chose to rest with little movement). Supine position for delivery was used routinely at all four hospitals; this was consistent with women's preferred choice of position, although very few women are aware of other positions. Qualitative findings suggest obstetricians and midwives favoured confining to bed during the first stage of labour, and supine position for delivery. CONCLUSIONS: The barriers to change appear to be complicated and require providers to want to change, and women to be informed of alternative positions during the first stage of labour and delivery. We believe that highlighting the gap between actual practice and current evidence provides a platform for dialogue with providers to evaluate the threats and opportunities for changing practice. PMID- 15113447 TI - Hyponatremic hypertensive syndrome (HHS) in an 18-month old-child presenting as malignant hypertension: a case report. AB - BACKGROUND: The combination of hyponatremia and renovascular hypertension is called hyponatremic hypertensive syndrome (HHS). Malignant hypertension as a presentation has been reported in adults with HHS but is rare in children. CASE PRESENTATION: An eighteen month-old male presented with drowsiness, sudden onset status epilepticus and blood pressure of 210/160. The electrolytes on admission revealed sodium of 120 mEq/L and potassium of 2.1 mEq/L. The peripheral renin activity (PRA) was 172 ng/ml/min (normal 3-11 ng/ml/min) and serum aldosterone level was 91 ng/dl (normal 4 to 16 ng/dl). Patient underwent angioplasty with no success, followed by surgical correction. Two years since the diagnosis, the blood pressure is controlled with labetolol and amlodipine (at less than sixth of the pre-operative dosages). The PRA is 2.4 ng/ml/min and aldosterone 15.5 ng/dl. The child not only had three renal arteries on left but all of them were stenosed which to best of our knowledge has not been described. CONCLUSION: As uncommon as HHS with malignant hypertension may be in adults it is under-reported in children and purpose of the case report is to raise its awareness. PMID- 15113448 TI - Assessing anti-rabies baiting--what happens on the ground? AB - BACKGROUND: Rabies is one of the most hazardous zoonoses in the world. Oral mass vaccination has developed into the most effective management method to control fox rabies. The future need to control the disease in large countries (i.e. Eastern Europe and the Americas) forces cost-benefit discussions. The 'Increase bait density' option refers to the usual management assumption that more baits per km2 could compensate for high fox abundance and override the imperfect supply of bait pieces to the individual fox. METHODS: We use a spatial simulation, which combines explicitly fox space use (tessellation polygons) and aeroplane flight lines (straight lines). The number of baits actually falling into each polygon is measured. The manager's strategic options are converted into changes of the resulting bait distribution on the ground. The comparison enables the rating of the options with respect to the management aim (i.e. accessibility of baits). RESULTS: Above 5% (approx. 10%) of all fox groups without any bait (at most 5 baits) relate to the baiting strategy applied in the field (1 km spaced parallel flight lines, 20 baits per km2 distributed) under habitat conditions comparable to middle and western Europe (fox group home-range 1 km2, 2.5 adults; reference strategy). Increasing the bait density on the same flight-line pattern neither reduces the number of under-baited fox group home-ranges, nor improves the management outcome and hence wastes resources. However, reducing the flight line distance provides a more even bait distribution and thus compensates for missed fox groups or extra high fox density.The reference strategy's bait density can be reduced when accounting for the missed fox groups. The management result with the proper strategy is likely the same but with reduced costs. CONCLUSION: There is no overall optimal strategy for the bait distribution in large areas. For major parts of the landscape, the reference strategy will be more competitive. In situations where set backs are attributed to non-homogeneous bait accessibility the distribution scheme has to be refined zone-based (i.e. increase of the flight line length per unit area). However, increase in bait density above the reference strategy appears inappropriate at least for non-urban abundance conditions of the red fox. PMID- 15113449 TI - Seroepidemiology of Herpes Simplex virus type 1 and 2 in Western and Southern Switzerland in adults aged 25-74 in 1992-93: a population-based study. AB - BACKGROUND: Genital herpes is one of the most prevalent sexually-transmitted diseases, and accounts for a substantial morbidity. Genital herpes puts newborns at risk for very severe disease and also increases the risk of horizontal HIV transmission. It thus stands as an important public health problem. The recent availability of type-specific gG-based assays detecting IgG against HSV-1 and HSV 2 allows to establish the prevalence of each subtype. Worldwide, few data have been published regarding the seroprevalence in general populations of HSV-2, the major causative agent for genital herpes, while no data exist regarding the Swiss population. METHODS: To evaluate the prevalence of IgG antibodies against HSV-1 and HSV-2 in Switzerland, we used a population-based serum repository from a health examination survey conducted in the Western and Southern area of Switzerland in 1992-93. A total of 3,120 sera were analysed by type-specific gG based ELISA and seroprevalence was correlated with available volunteers characteristics by logistic regression. RESULTS: Overall, seroprevalence rates were 80.0 +/- 0.9% (SE, 95% CI: 78.1-81.8) for HSV-1 and 19.3 +/- 0.9% (SE, 95% CI: 17.6-21.1) for HSV-2 in adults 35-64 year old. HSV-1 and HSV-2 seroprevalence increased with age, with a peak HSV-2 seroprevalence in elderly gentlemen, possibly a seroarcheological evidence of sexually transmitted disease epidemics during World War II. Risk factors for HSV-2 infection included female sex, marital status other than married, and size of town of residence larger than 1500 inhabitants. Unexpectedly and conversely to HSV-1, HSV-2 seroprevalence increased with educational level. HSV-2 infection was less prevalent among HSV-1 infected individuals when compared to HSV-1 uninfected individuals. This effect was most apparent among women at high risk for HSV-2 infection. CONCLUSIONS: Our data demonstrate that by the early nineties, HSV-2 had spread quite largely in the Swiss population. However, the epidemiology of HSV-2 in Switzerland presents paradoxical characteristics, e.g. positive correlation with education level, that have not been observed elsewhere. PMID- 15113451 TI - Lactobacillus GG in inducing and maintaining remission of Crohn's disease. AB - BACKGROUND: Experimental studies have shown that luminal antigens are involved in chronic intestinal inflammatory disorders such as Crohn's disease and ulcerative colitis. Alteration of the intestinal microflora by antibiotic or probiotic therapy may induce and maintain remission. The aim of this randomized, placebo controlled trial was to determine the effect of oral Lactobacillus GG (L. GG) to induce or maintain medically induced remission. METHODS: Eleven patients with moderate to active Crohn's disease were enrolled in this trial to receive either L. GG (2 x 10(9) CFU/day) or placebo for six months. All patients were started on a tapering steroid regime and received antibiotics for the week before the probiotic/placebo medication was initiated. The primary end point was sustained remission, defined as freedom from relapse at the 6 months follow-up visit. Relapse was defined as an increase in CDAI of >100 points. RESULTS: 5/11 patients finished the study, with 2 patients in each group in sustained remission. The median time to relapse was 16 +/- 4 weeks in the L. GG group and 12 +/- 4.3 weeks in the placebo group (p = 0.5). CONCLUSION: In this study we could not demonstrate a benefit of L. GG in inducing or maintaining medically induced remission in CD. PMID- 15113452 TI - The role of guidelines and the patient's life-style in GPs' management of hypercholesterolaemia. AB - BACKGROUND: Recent Swedish and joint European guidelines on hyperlipidaemia stress the high coronary risk for patients with already established arterio sclerotic disease (secondary prevention) or diabetes. For the remaining group, calculation of the ten-year risk for coronary events using the Framingham equation is suggested. There is evidence that use of and adherence to guidelines is incomplete and that tools for risk estimations are seldom used. Intuitive risk estimates are difficult and systematically biased. The purpose of the study was to examine how GPs use knowledge of guidelines in their decisions to recommend or not recommend a cholesterol-lowering drug and the reasons for their decisions. METHODS: Twenty GPs were exposed to six case vignettes presented on a computer. In the course of six screens, successively more information was added to the case. The doctors were instructed to think aloud while processing the cases (Think-Aloud Protocols) and finally to decide for or against drug treatment. After the six cases they were asked to describe how they usually reason when they meet patients with high cholesterol values (Free-Report Protocols). The two sets of protocols were coded for cause-effect relations that were supposed to reflect the doctors' knowledge of guidelines. The Think-Aloud Protocols were also searched for reasons for the decisions to prescribe or not to prescribe. RESULTS: According to the protocols, the GPs were well aware of the importance of previous coronary heart disease and diabetes in their decisions. On the other hand, only a few doctors mentioned other arterio-sclerotic diseases like stroke and peripheral artery disease as variables affecting their decisions. There were several instances when the doctors' decisions apparently deviated from their knowledge of the guidelines. The arguments for the decisions in these cases often concerned aspects of the patient's life-style like smoking or overweight- either as risk increasing factors or as alternative strategies for intervention. CONCLUSIONS: Coding verbal protocols for knowledge and for decision arguments seems to be a valuable tool for increasing our understanding of how guidelines are used in the on treatment of hypercholesterolaemia. By analysing arguments for treatment decisions it was often possible to understand why departures from the guidelines were made. While the need for decision support is obvious, the current guidelines may be too simple in some respects. PMID- 15113453 TI - Impact of disaster-related mortality on gross domestic product in the WHO African Region. AB - BACKGROUND: Disaster-related mortality is a growing public health concern in the African Region. These deaths are hypothesized to have a significantly negative effect on per capita gross domestic product (GDP). The objective of this study was to estimate the loss in GDP attributable to natural and technological disaster-related mortality in the WHO African Region. METHODS: The impact of disaster-related mortality on GDP was estimated using double-log econometric model and cross-sectional data on various Member States in the WHO African Region. The analysis was based on 45 of the 46 countries in the Region. The data was obtained from various UNDP and World Bank publications. RESULTS: The coefficients for capital (K), educational enrolment (EN), life expectancy (LE) and exports (X) had a positive sign; while imports (M) and disaster mortality (DS) were found to impact negatively on GDP. The above-mentioned explanatory variables were found to have a statistically significant effect on GDP at 5% level in a t-distribution test. Disaster mortality of a single person was found to reduce GDP by US$0.01828. CONCLUSIONS: We have demonstrated that disaster related mortality has a significant negative effect on GDP. Thus, as policy makers strive to increase GDP through capital investment, export promotion and increased educational enrolment, they should always keep in mind that investments made in the strengthening of national capacity to mitigate the effects of national disasters expeditiously and effectively will yield significant economic returns. PMID- 15113454 TI - ATF3 upregulation in glia during Wallerian degeneration: differential expression in peripheral nerves and CNS white matter. AB - BACKGROUND: Many changes in gene expression occur in distal stumps of injured nerves but the transcriptional control of these events is poorly understood. We have examined the expression of the transcription factors ATF3 and c-Jun by non neuronal cells during Wallerian degeneration following injury to sciatic nerves, dorsal roots and optic nerves of rats and mice, using immunohistochemistry and in situ hybridization. RESULTS: Following sciatic nerve injury--transection or transection and reanastomosis--ATF3 was strongly upregulated by endoneurial, but not perineurial cells, of the distal stumps of the nerves by 1 day post operation (dpo) and remained strongly expressed in the endoneurium at 30 dpo when axonal regeneration was prevented. Most ATF3+ cells were immunoreactive for the Schwann cell marker, S100. When the nerve was transected and reanastomosed, allowing regeneration of axons, most ATF3 expression had been downregulated by 30 dpo. ATF3 expression was weaker in the proximal stumps of the injured nerves than in the distal stumps and present in fewer cells at all times after injury. ATF3 was upregulated by endoneurial cells in the distal stumps of injured neonatal rat sciatic nerves, but more weakly than in adult animals. ATF3 expression in transected sciatic nerves of mice was similar to that in rats. Following dorsal root injury in adult rats, ATF3 was upregulated in the part of the root between the lesion and the spinal cord (containing Schwann cells), beginning at 1 dpo, but not in the dorsal root entry zone or in the degenerating dorsal column of the spinal cord. Following optic nerve crush in adult rats, ATF3 was found in some cells at the injury site and small numbers of cells within the optic nerve displayed weak immunoreactivity. The pattern of expression of c-Jun in all types of nerve injury was similar to that of ATF3. CONCLUSION: These findings raise the possibility that ATF3/c-Jun heterodimers may play a role in regulating changes in gene expression necessary for preparing the distal segments of injured peripheral nerves for axonal regeneration. The absence of the ATF3 and c-Jun from CNS glia during Wallerian degeneration may limit their ability to support regeneration. PMID- 15113455 TI - Abnormal auditory mismatch response in tinnitus sufferers with high-frequency hearing loss is associated with subjective distress level. AB - BACKGROUND: Tinnitus is an auditory sensation frequently following hearing loss. After cochlear injury, deafferented neurons become sensitive to neighbouring intact edge-frequencies, guiding an enhanced central representation of these frequencies. As psychoacoustical data 123 indicate enhanced frequency discrimination ability for edge-frequencies that may be related to a reorganization within the auditory cortex, the aim of the present study was twofold: 1) to search for abnormal auditory mismatch responses in tinnitus sufferers and 2) relate these to subjective indicators of tinnitus. RESULTS: Using EEG-mismatch negativity, we demonstrate abnormalities (N = 15) in tinnitus sufferers that are specific to frequencies located at the audiometrically normal lesion-edge as compared to normal hearing controls (N = 15). Groups also differed with respect to the cortical locations of mismatch responsiveness. Sources in the 90-135 ms latency window were generated in more anterior brain regions in the tinnitus group. Both measures of abnormality correlated with emotional-cognitive distress related to tinnitus (r approximately .76). While these two physiological variables were uncorrelated in the control group, they were correlated in the tinnitus group (r =.72). Concerning relationships with parameters of hearing loss (depth and slope), slope turned out to be an important variable. Generally, the steeper the hearing loss is the less distress related to tinnitus was reported. The associations between slope and the relevant neurophysiological variables are in agreement with this finding. CONCLUSIONS: The present study is the first to show near-to-complete separation of tinnitus sufferers from a normal hearing control group based on neurophysiological variables. The finding of lesion-edge specific effects and associations with slope of hearing loss corroborates the assumption that hearing loss is the basis for tinnitus development. It is likely that some central reorganization follow a damage to hearing receptors, even though the paradoxical results indicate that they most likely are somewhat different than originally assumed (see Background). One partial explanation might lie in the involvement of top-down (presumably frontal-lobe) controlled processes. A better comprehension of the exact mechanisms leading to the present results could have a broad impact on the understanding and perhaps treatment of tinnitus. PMID- 15113456 TI - A novel medium for the enhanced cell growth and production of prodigiosin from Serratia marcescens isolated from soil. AB - BACKGROUND: Prodigiosin produced by Serratia marcescens is a promising drug owing to its reported characteristics of having antifungal, immunosuppressive and antiproliferative activity. From an industrial point of view the necessity to obtain a suitable medium to simultaneously enhance the growth of Serratia marcescens and the pigment production was the aim of this work. The usage of individual fatty acid as substrate in industries would be cost-effective in the long run and this paved the way for us to try the effect of different fatty acid containing seeds and oils of peanut, sesame and coconut as source of substrate. RESULTS: The addition of sugars only showed slight enhancement of prodigiosin production in nutrient broth but not in fatty acid containing seed medium. The powdered peanut broth had supported better growth of Serratia marcescens and higher yield of prodigiosin when compared with the existing nutrient broth and peptone glycerol broth. A block in prodigiosin production was seen above 30 degrees C in nutrient broth, but the fatty acid seed medium used by us supported prodigiosin production upto 42 degrees C though the yields were lower than what was obtained at 28 degrees C. From the results, the fatty acid form of carbon source has a role to play in enhanced cell growth and prodigiosin production. CONCLUSION: We conclude by reporting that the powdered and sieved peanut seed of different quality grades were consistent in yielding a fourty fold increase in prodigiosin production over the existing media. A literature survey on the composition of the different media components in nutrient broth, peptone glycerol broth and the fatty acid containing seeds and oils enabled us to propose that the saturated form of fatty acid has a role to play in enhanced cell growth and prodigiosin production. This work has also enabled us to report that the temperature related block of prodigiosin biosynthesis varies with different media and the powdered peanut broth supports prodigiosin production at higher temperatures. The medium suggested in this work is best suitable from an industrial point of view in being economically feasible, in terms of the higher prodigiosin yield and the extraction of prodigiosin described in this paper is simple with minimal wastage. PMID- 15113457 TI - Endoscopy and transsphenoidal surgery. AB - ENDOSCOPY OFFERS INTERNAL visualization of many different cavities of the human body, with its specific vision inside the anatomy, close to the target area. The view of the surgical field in transsphenoidal surgery had been obtained with the naked eye from its beginning in 1907 up to the introduction of the operating microscope by Jules Hardy in the 1960s, which represented a great advance in terms of magnification and illumination. In the past decade, modern rigid endoscopes, with their wider view near the relevant anatomy, have permitted minimally traumatic transsphenoidal procedures in and around the sellar area, thus representing a "new wave" in transsphenoidal history. An overview of the evolution of the endoscope as a visualizing and operating instrument particularly related to the transsphenoidal approach is presented here. The current possibilities of transsphenoidal endoscopy, with its related advantages and limitations, are presented. PMID- 15113458 TI - Tumor-third ventricular relationships in supradiaphragmatic craniopharyngiomas: correlation of morphological, magnetic resonance imaging, and operative findings. AB - OBJECTIVE: To disclose the relationships of primarily supradiaphragmatic craniopharyngiomas with the third ventricular floor (3rdVF) by means of preoperative magnetic resonance imaging (MRI) and thus to select the surgical approach avoiding the hypothalamic structures. METHODS: MRI findings in 76 consecutive patients with craniopharyngiomas operated on between June 1991 and December 2002 were interpreted on the basis of the results of the authors' own previous microanatomic studies. The assumed tumor-3rdVF relationships were then correlated with the operative findings. MRI features characteristic for different topographical relationships were analyzed in 44 patients (18 children, 26 adults) with exclusively supradiaphragmatic tumors. RESULTS: In 14 of 15 patients with the tumor located below the 3rdVF (suprasellar extraventricular craniopharyngioma), the anterior communicating artery was displaced upward and indirectly indicated the position of the chiasm between the prechiasmatic and the retrochiasmatic tumor portions. Hydrocephalus was absent in 14 patients, including those with giant tumors. The anterior part of the third ventricular cavity was found in front of the level of the foramina of Monro in 6 patients. All 28 tumors growing partially inside and partially outside the third ventricular cavity (intraventricular and extraventricular craniopharyngioma) were retrochiasmatic. They caused severe or moderate hydrocephalus in 20 patients and mild hydrocephalus in 2. One purely intraventricular tumor caused severe hydrocephalus. CONCLUSION: The position of the optic chiasm and the size of the lateral ventricles on preoperative MRI enable us to determine the position of the 3rdVF or its remnants in relation to the supradiaphragmatic craniopharyngiomas and to select the proper surgical approach allowing exposure of the tumor while avoiding the hypothalamic structures. PMID- 15113459 TI - Motor evoked potential monitoring with supratentorial surgery. AB - OBJECTIVE: To assess feasibility and clinical value of motor evoked potential (MEP) monitoring with surgery close to supratentorial motor areas and pathways. METHODS: Functional mapping by somatosensory evoked potential phase reversal and continuous MEP recording after high-frequency repetitive electrical cortex stimulation was performed during 182 operations in 177 patients. Significant MEP changes were reported to trigger surgical reaction. Intraoperative surgical and electrophysiological findings were documented prospectively. Patient files were reviewed for clinical data. MEP monitoring results were correlated with motor outcome. RESULTS: MEP recording was successful in 167 cases (91.8%). Inadequate electrode placement was an important reason for failed recording in the remaining patients, whereas preoperative paresis and anesthesia had no significant effect. Permanently disabling new motor deficit occurred in 8 cases (4.9%), whereas transient and nondisabling weakness was frequent (27.4%). Significant MEP changes occurred during 64 operations (39%). Irreversible MEP loss always predicted new, usually permanent, paresis. Unaltered MEP recordings indicated unimpaired motor function in the monitored muscle groups, except for rare transient deficit because of late edema and rebleeding. Irreversible MEP deterioration without loss and reversible changes could be associated with new paresis, which was transient in most patients. No major complications were observed, except for intraoperative generalized seizure in one epilepsy patient under insufficient anticonvulsant therapy. CONCLUSION: MEP monitoring with supratentorial surgery is feasible and safe. It may help to maximize resection within the limits of preserved motor function. Further evidence is needed to confirm these results. PMID- 15113460 TI - Duration of cognitive impairment after sports concussion. AB - OBJECTIVE: We sought to determine the duration of cognitive impairment after sports concussion. METHODS: We conducted a study with a prospective design in which 729 athletes underwent preseason baseline testing by being administered a computerized neuropsychological test battery, followed by retesting at regular intervals after they sustained sports-related concussions. A control group consisting of nonconcussed athletes drawn from the same baseline population underwent testing at parallel intervals. RESULTS: Cognitive impairment in this primarily American Academy of Neurology Grade II sample of boxing concussions was apparent on the day of injury and at 1 to 2 days postinjury. Recovery of cognitive performance occurred during the 3- to 7-day interval. Comparison with control subjects showed that absent or attenuated practice effects, in addition to frank deterioration from baseline, were indications of recent concussion. CONCLUSION: The present findings of recovery during the 3- to 7-day interval postinjury are consistent with the American Academy of Neurology Grade II return to-play practice parameters suggesting a 1-week time-out from participation in contact sports. PMID- 15113461 TI - Resource use after subarachnoid hemorrhage: comparison between endovascular and surgical treatment. AB - OBJECTIVE: The aim of this study was to compare resource use after endovascular treatment and surgical clipping of ruptured intracranial aneurysms. METHODS: The study population consisted of patients with aneurysmal subarachnoid hemorrhage who were actively treated in Kuopio University Hospital. The baseline patients' characteristics were obtained from the institution's aneurysm registry. Variables indicating resource use in the intensive care unit (ICU) were obtained from the ICU patient data management system. The Therapeutic Intervention Scoring System was used to measure the intensity of treatment in the ICU. All care days in the ICU and in the hospital and all Therapeutic Intervention Scoring System points that were accumulated during the 12-month follow-up period were calculated. Outcomes were assessed according to the Glasgow Outcome Scale. RESULTS: The study population consisted of 68 patients who underwent endovascular treatment and 103 patients who had surgery. The median lengths of stay in the ICU (1.7 versus 1.8 d) and the hospital (14.0 versus 15.0 d), as well as the accumulated Therapeutic Intervention Scoring System points (56 versus 55), among patients who underwent endovascular or surgical treatment were similar (P = NS for all). The modality of treatment did not influence the number of ICU or hospital patient days or the intensity of ICU treatment, regardless of the patient's preoperative clinical status. CONCLUSION: The modality of treatment of patients with subarachnoid hemorrhage does not seem to affect resource use. Endovascular and surgical treatment are likely to require a similar amount of ICU resources in the year after initial treatment. PMID- 15113462 TI - Strategies for the management of intraprocedural thromboembolic complications with abciximab (ReoPro). AB - OBJECTIVE: Most complications related to endovascular neurointerventional procedures are thromboembolic. The objective of this study was to determine whether abciximab, a glycoprotein IIb/IIIa inhibitor, is a safe and effective agent for the treatment of thromboembolic complications encountered during interventional neuroradiological procedures. METHODS: A search of a prospectively maintained database identified 13 patients who were treated for intraprocedural thromboembolic complications with abciximab, either alone or in combination with tissue plasminogen activator, during a 22-month period. RESULTS: Of the 13 patients in our series (10 with aneurysms, 3 with arteriovenous malformations), 12 presented with unsecured vascular lesions, and in 9 cases the lesions were either acutely (<10 d) or subacutely (<6 mo) hemorrhagic. After the identification of a thromboembolic complication, each patient was treated with either intra-arterial (n = 5) or intravenous (n = 8) abciximab. Abciximab was administered intravenously as a 0.25-mg/kg bolus (n = 2), a 0.25-mg/kg bolus followed by a 12-hour 0.125-microg/kg/min infusion (n = 5), or a 12-hour 0.125 microg/kg/min infusion alone (n = 1). Intra-arterial abciximab (3.5-10 mg) was administered directly through a microcatheter positioned in the vicinity of the thrombus (n = 5). Five patients also were administered alteplase. Complete (n = 7) or partial (n = 6) resolution of thrombus was observed in all cases. Five patients had small infarcts in the distribution of the thromboembolic complication. In no case was new or increased hemorrhaging observed after thrombolysis. CONCLUSION: Abciximab (ReoPro), administered either intravenously or intra-arterially, is a viable option for the management of thromboembolic complications encountered during endovascular therapeutic procedures. PMID- 15113463 TI - Prevention of pulmonary embolism by combined modalities of thromboprophylaxis and intensive surveillance protocol. AB - OBJECTIVE: Deep vein thrombosis (DVT) is a known complication in neurosurgical patients. The protocols for the prevention of DVT and pulmonary embolism (PE) are not applied universally. Our goal was to review the incidence of DVT and PE in neurosurgical intensive care units (NSICUs) and to compare it with the incidence of DVT and PE in other intensive care units (ICUs) in the same hospital. METHODS: We retrospectively reviewed 5327 consecutive admissions in all intensive care units at the University of Illinois Hospital between July 2001 and June 2002. The NSICU had 1094 admissions, and all other ICUs combined had 4233 admissions. In our NSICU, every patient was administered DVT prophylaxis in the form of subcutaneous heparin and a sequential compression device if these treatments were not contraindicated. Lower-extremity venous Doppler ultrasonography was performed twice weekly in all NSICU patients. There was no routine use of DVT prophylaxis in the other ICUs, and no other ICUs performed routine lower-extremity venous Doppler ultrasonography. We compared the incidence of DVT and PE in the NSICU and the combined population of other adult ICUs. RESULTS: PE was diagnosed in 1 (0.09%) of 1094 patients in the NSICU and in 45 (1.06%) of 4233 patients in all other ICUs (P < 0.0001, Z = 5.34). DVT was diagnosed in 28 patients (2.55%) in the NSICU and in 238 patients (5.62%) in all other ICUs (P < 0.001, Z = 4.32). These values suggest a statistically significant difference in the incidence of DVT and PE in the NSICU compared with all other ICUs. CONCLUSION: We conclude that there was a significantly lower incidence of DVT and PE in the NSICU than in all other ICUs at our institution. DVT prophylaxis and twice-weekly lower extremity venous Doppler screening in the NSICU have been found to be beneficial in decreasing the incidence of DVT and particularly effective in preventing PE. PMID- 15113464 TI - Study of magnetic resonance imaging-based arteriovenous malformation delineation without conventional angiography. AB - OBJECTIVE: In this study, we aimed to assess the feasibility of arteriovenous malformation (AVM) delineation for gamma knife radiosurgery without conventional angiography and to correlate factors that may affect AVM delineation. METHODS: A series of 57 consecutive patients with AVMs treated with gamma knife radiosurgery from August 1994 to December 2000 were reviewed. All patients in the study had undergone pretreatment angiography. The mean AVM volume was 2.8 cm(3), with a median of 2.0 cm(3) (range, 0.04-22 cm(3)). All AVMs were delineated on the original frame-based magnetic resonance imaging (MRI) scans by a vascular neurosurgeon without the assistance of angiography and then compared with the actual AVM delineation on the basis of previously performed angiography and MRI. Univariate correlation analysis was used to determine the relationship of AVM coverage, size, diffuseness, previous embolization, and hemorrhage parameters. RESULTS: The study volume or MRI-based volume alone coincided with the actual treatment volume by a mean of 58% for diffuse and 87% for nondiffuse AVMs (P = 0.0005). At AVM volume greater than 2 cm(3), the median percentage of coinciding volume was 63% for embolized AVMs and 82% for nonembolized AVMs (P = 0.0315). Conversely, the study volume overestimated the actual treatment volume by a mean of 57% for AVMs larger than 2 cm(3) versus 25% for AVMs smaller than 2 cm(3) (P = 0.0012). In general, the percentage of the coinciding volume was inversely related to that of the excess volume, whereas both the study volume and the coinciding volume were proportionate to AVM volume at treatment. CONCLUSION: MRI based AVM delineation without conventional angiography may be feasible only for selected patients, such as those with nondiffuse and large nonembolized AVMs. PMID- 15113465 TI - Associations among magnetic resonance spectroscopy, apparent diffusion coefficients, and image-guided histopathology with special attention to radiation necrosis. AB - OBJECTIVE: In patients with malignant glioma previously treated with surgery, radiation, and chemotherapy, clinical and radiographic signs of recurrent disease often require differentiation between radiation necrosis and recurrent tumor. Published work suggests that although magnetic resonance spectroscopy (MRS) can reliably differentiate pure tumor, pure necrosis, and spectroscopically normal tissues, it may not be particularly helpful because most patients have mixed histological findings comprised of necrosis and tumor. To improve our clinical ability to discriminate among these histological entities, we have analyzed MRS in conjunction with apparent diffusion coefficient (ADC) sequences derived from magnetic resonance imaging. METHODS: In 18 patients, spectroscopic and diffusion weighted images were obtained before surgery for suspected recurrent neoplastic disease. Spectral data for pure tumor, pure necrosis, and mixed tumor and necrosis were derived from 65 spectroscopic observations in patients with previously treated gliomas (n = 16) and metastatic tumors (n = 2). Spectral data for choline (Cho), N-acetylaspartate (NAA), creatine (Cr), and lipid-lactate were analyzed separately and in conjunction with ADCs in all patients (15 observations of pure tumor, 33 observations of pure necrosis, and 13 observations of mixed tumor and necrosis). Histological specimens were obtained stereotactically at the time of surgery (<48 h after image acquisition) for recurrent disease and digitally co-registered with MRS data. RESULTS: ADC values for pure tumor, pure necrosis, and mixed tumor and necrosis were 1.30, 1.60, and 1.42, respectively. Cho/NAA less than 0.20, NAA/normal Cr greater than 1.56, and NAA/Cho greater than 1.32 increase the odds that a tissue biopsy will be pure necrosis versus mixed tumor and necrosis. Although various values of all MRS ratios analyzed may provide positive correlations for histopathological differentiation of tissue between that of pure tumor and that of pure necrosis, the addition of ADC values to only NAA/Cho and NAA/normal Cr increases the odds of correct differentiation between pure tumor and pure necrosis. The addition of ADC values does not provide additional information beyond that of MRS in distinguishing specimens of mixed tumor and necrosis from either pure tumor or pure necrosis. CONCLUSION: It has been demonstrated that MRS ratio analysis may allow for the clinical discrimination between specimens of pure tumor and pure necrosis, and the addition of ADC data into this analysis may enhance this specific differentiation. However, although a trend toward correlation between ADC values and the various histopathological features was noted, the direct addition of ADC data does not seem to allow further discrimination, beyond that provided by MRS, among specimens of mixed tumor and necrosis and either pure tumor or pure necrosis. PMID- 15113466 TI - Thalamic deep brain stimulation for essential tremor: relation of lead location to outcome. AB - OBJECTIVE: Thalamic deep brain stimulation (DBS) is commonly used to treat essential tremor, but the optimal lead location within the thalamus has not been systematically evaluated. We examined the relation of lead location to clinical outcome in a series of essential tremor patients treated by thalamic DBS. METHODS: Fifty-seven leads in 37 patients were studied. Lead locations were measured by postoperative magnetic resonance imaging. Contralateral arm tremor was assessed in the DBS-on and DBS-off states using the Fahn-Tolosa-Marin tremor rating scale, with a mean follow-up of 26 months. Lead locations were statistically correlated, using analysis of variance, with percent improvement in tremor resulting from DBS activation. RESULTS: Improvement in tremor score was significantly correlated with lead location in both the anteroposterior and lateral dimensions. In the plane of the commissures, the optimal electrode location was determined statistically to be 6.3 mm anterior to the posterior commissure and 12.3 mm lateral to the midline, or 10.0 mm lateral to the third ventricle. CONCLUSION: Optimal electrode location for thalamic DBS in essential tremor corresponds to the anterior margin of the ventralis intermedius nucleus. Leads located greater than 2 mm (in the plane of the commissures) from the optimal coordinates are more likely to be associated with poor tremor control than leads within 2 mm of the optimal location. The incidence of true physiological tolerance to the antitremor effect of thalamic DBS (defined as poor tremor control in spite of lead location within 2 mm of the optimal site) was found to be 9%. PMID- 15113467 TI - Intraoperative Iso-C C-arm navigation in craniospinal surgery: the first 60 cases. AB - OBJECTIVE: The intraoperative Iso-C C-arm (Siremobil Iso-C 3D; Siemens Medical Solutions, Erlangen, Germany) provides a unique ability to acquire and view multiplanar three-dimensional images of intraoperative anatomy. Registration for intraoperative surgical navigation may be automated, thus simplifying the operative workflow. METHODS: Iso-C C-arm intraoperative fluoroscopy acquires 100 images, each of which must be 1.8 degrees in a circumferential fashion about an "isocentric" point in space. The system generates a high-resolution isotropic three-dimensional data set that is available immediately after the 90-second C arm rotation. The data set is ported to the image-guided workstation, registration is immediate and automated, and the surgeon can navigate with millimetric accuracy. The authors prospectively examined data from the initial 60 patients examined with the Iso-C, among whom were cases of anterior and posterior spinal instrumentation from the occiput to the sacrum. Percutaneous and minimally invasive spinal and cranial procedures were also included. RESULTS: Automated registration for image-guided navigation was attainable for anterior and posterior cases from the cranial base and entire spine. In most cases, intraoperative postprocedural imaging with the Iso-C mitigated the need for postoperative imaging. CONCLUSION: Intraoperative Iso-C three-dimensional scanning allows real-time feedback during cranial base and spinal surgery and during procedures involving instrumentation. In most cases, it obviates the need for postoperative computed tomography. Its usefulness is in its simplicity, and it can be easily adapted to the operating room workflow. When coupled with intraoperative navigation, this new technology facilitates complex neurosurgical procedures by improving the accuracy, safety, and time of surgery. PMID- 15113468 TI - Reliability of three-dimensional fluoroscopy for detecting pedicle screw violations in the thoracic and lumbar spine. AB - OBJECTIVE: Thoracic and lumbar pedicle screws have become popular because of their biomechanical superiority over other methods of spinal fixation. However, the safety and efficacy of transpedicular screws depend on their proper placement. Recent advances in imaging have resulted in the ability to acquire three-dimensional (3-D) axial images of the spine during surgery, and this study was undertaken to assess the reliability of this technology to detect pedicle violations. METHODS: Pedicle screws were placed in six human cadaver spines from T1 to S1 using standard techniques. Intentional pedicle violations were created in 74 of 216 pedicles, and violations were graded on a four-point scale (range, 0 3). Radiographic images were then obtained using a conventional spiral computed tomographic scanner and the Siremobil Iso-C 3D (Siemens Medical Solutions, Erlangen, Germany) 3-D fluoroscopy unit. An independent neuroradiologist then graded pedicle violations as ascertained by the two imaging modalities. RESULTS: Using direct inspection of the pedicles as the "gold standard," the overall sensitivity and specificity for detecting pedicle violations were 0.716 and 0.789, respectively, with 3-D fluoroscopy. The overall sensitivity and specificity for detecting pedicle violations were 0.608 and 0.937, respectively, with conventional computed tomography. All Grade 2 pedicle violations were detected in the thoracic spine by both modalities, and all Grade 3 violations were detected by both modalities. CONCLUSION: Axial images obtained with 3-D fluoroscopy demonstrate a higher sensitivity but lower specificity than conventional computed tomographic scanning for assessing pedicle violations. By providing real-time intraoperative imaging, 3-D fluoroscopy may enhance the safety of thoracic transpedicular instrumentation. PMID- 15113469 TI - The anterior surgical approach to the cervical spine for intervertebral disc disease. AB - THE ANTERIOR SURGICAL APPROACH to the cervical spine in patients with discogenic compressive pathological findings causing radiculopathy or myelopathy is a commonly performed operation with several technical variations. We describe the normal and pathological anatomy and the techniques of surgical decompression of the dura with autograft fusion, which we have used for the past 35 years. PMID- 15113470 TI - Thoracoscopic placement of dual-rod instrumentation in thoracic spinal trauma. AB - OBJECTIVE AND IMPORTANCE: Traditionally, thoracic fractures that require anterior stabilization are treated through an open thoracotomy approach. Thoracoscopic instrumentation avoids many of the complications associated with an open thoracotomy but is technically challenging. We report the first cases of dual-rod internal fixation systems placed thoracoscopically for thoracic spinal trauma. CLINICAL PRESENTATION: Two male patients sustained midthoracic spinal trauma falling from motorcycles in separate incidents. Both injuries led to unstable spinal columns, but the patients had no neurological deficits and had minimal spinal cord compression. One patient had a complex spiral fracture from T6 to T8; the other had T7 burst and T8 compression fractures. Based on the complex morphological features of the patients' fractures, anterior internal fixation was the treatment of choice for both. The two available options for an anterior stabilization were open thoracotomy and thoracoscopic instrumentation. Because extensive decompression was unnecessary, a thoracoscopic approach was used. INTERVENTION: A dual-rod internal fixation system (Medtronic Sofamor Danek, Inc., Memphis, TN) was placed with two screws each in the T6 and T9 vertebral bodies of each patient. Thoracoscopy was used for direct visualization of the operative site with fluoroscopic guidance for screw placement. Surgery was completed without complications, and both patients did well afterward. Upright and supine x rays demonstrated that the constructs were stable at 10 weeks and 6 months, respectively. CONCLUSION: Thoracoscopic instrumentation offers the advantages of a minimally invasive approach but is technically challenging. The characteristics of dual-rod fixation systems (small-profile components and step-wise insertion) provide the best biomechanical profile and facilitate thoracoscopic instrumentation. PMID- 15113471 TI - Median nerve excursion during endoscopic carpal tunnel release. AB - OBJECTIVE: Restriction of the excursion of the nerve has been accepted as a pathogenetic element in carpal tunnel syndrome. The goal of this article was to evaluate the median nerve excursion in the carpal tunnel measured as a function of wrist position before and after endoscopic carpal tunnel release (ECTR) on 28 hands of 22 patients. METHODS: The position of cylindrical stainless steel markers embedded within the median nerve was measured by a direct radiographic technique. Each upper extremity was examined in three wrist positions. Then, endoscopic release with Menon's technique was performed, and the measurements were repeated. RESULTS: In this prospective clinical study, most (93%) of the patients experienced resolution of their symptoms. Before and after ECTR, median nerve excursion was linear and was affected by wrist position. Before ECTR, when the wrist was moved from the end of dorsiflexion to the end of palmar flexion, the median nerve underwent a mean total excursion of 28.8 mm at the wrist. A comparison of the before and after ECTR excursion showed no statistical differences in the amount of motion. CONCLUSION: The single-portal ECTR does not seem to influence the median nerve excursion for the wrist positions studied in patients with carpal tunnel syndrome. The results from this in vivo study showed longitudinal gliding of the median nerve twice as great as in in vitro studies. PMID- 15113472 TI - A combined subtemporal and transventricular/transchoroidal fissure approach to medial temporal lesions. AB - OBJECTIVE: To minimize therapeutic morbidity such as cognitive function disturbance and vascular injury to perforating arteries, preoperative functional mapping of the basal temporal lobe functions was performed and the medial temporal lesions were resected via a combined subtemporal and transventricular/transchoroidal fissure approach. METHODS: Twenty-one patients with medial temporal lesions including tumors, arteriovenous malformations, and medial temporal lobe epilepsy underwent operation. The neurovascular structures in the ambient cistern were first dissected free from the medial temporal lobe with a conventional subtemporal approach. Then, the temporal horn was opened through the basal surface of the temporal lobe. Finally, the ambient cistern was accessed from the temporal horn through the choroidal fissure. In five patients whose lesions were revealed to be located on the dominant side by preoperative intracarotid amytal administration test (Wada test), functional mapping of the basal temporal lobe language cortex was monitored for 1 week by use of a subdural electrocorticogram grid before the extirpation surgery. The entrance point from the temporal base to the temporal horn was determined by the result of the functional mapping. RESULTS: The lesions were resected safely and completely in all cases. Language and cognitive functions were preserved even in patients with the basal language area on the dominant side. CONCLUSION: Surgeons can confirm the important neurovascular structures from the subtemporal route and from the transtemporal horn route by a combined subtemporal and transventricular/transchoroidal fissure approach. This approach is especially effective for avoiding ischemic complications by allowing direct confirmation of the anterior choroidal and thalamoperforating arteries. PMID- 15113473 TI - The open-mouth fronto-orbitotemporozygomatic approach for extensive benign tumors with coexisting splanchnocranial and neurocranial involvement. AB - OBJECTIVE: To describe the advantages of a modification of the standard fronto orbitotemporozygomatic (FOTZ) approach, consisting of the forced opening of the patient's mouth (open-mouth FOTZ), for the treatment of benign tumors massively involving the splanchnocranium and neurocranium. METHODS: The modified FOTZ approach obtained by forced mouth opening is described. Such a procedure was introduced with the aim of obtaining a minimally invasive access to lesions involving both the infratemporal-parapharyngeal spaces and the anteromedial cranial fossae. The forced opening of the mouth allows the surgeon to keep the coronoid process of the mandible away from the operating field, producing a wide exposure of the infratemporal space like that of the posterior wall of the maxillary sinus, which can be penetrated from behind without adding destructive procedures on the mandibular bone. RESULTS: From a series of 45 patients affected by cranial base lesions operated on during an 8-year time period, three patients affected by juvenile nasopharyngeal angiofibromas were selected for an open-mouth FOTZ approach. In all cases, a complete eradication of both the intracranial and extracranial components of the tumor was possible by this approach. The same procedure was then used in four patients affected by extensive meningiomas and two patients harboring Vth cranial nerve schwannomas, with complete tumor removal. CONCLUSION: In our experience, the open-mouth FOTZ approach seems particularly suited for extensive benign tumors (including juvenile nasopharyngeal angiofibromas) with splanchnocranial and neurocranial involvement. No indication exists for the use of this approach in malignant tumors, in which total eradication with large tumor-free margins is required. PMID- 15113474 TI - The carotid-oculomotor window in exposure of upper basilar artery aneurysms: a cadaveric morphometric study. AB - OBJECTIVE: The carotid-oculomotor window remains the traditional deep window in the exposure of aneurysms of the upper basilar artery. Although several techniques have been described to expand this window, few morphometric studies document either the degree of its expansion or its contribution to the exposure of the basilar artery. We review the microsurgical anatomy of the carotid oculomotor window, describe expansion techniques, and analyze morphometrically the contribution of each step (i.e., extradural anterior clinoidectomy, mobilization of the internal carotid artery [ICA], and posterior clinoidectomy) to the expansion of the window and/or exposure of the artery. METHODS: Ten formalin-fixed, alcohol-preserved, cadaver heads injected with pigmented silicone were prepared for bilateral dissection. The vertebrobasilar system was injected with pigmented silicone mixed with barium (1:1), rendering it radiopaque. After completing a frontotemporal-orbitozygomatic craniotomy, we performed dissection in two stages: Stage I consisted of a conventional transsylvian exposure of the upper basilar artery through the carotid-oculomotor window; and Stage II added anterior clinoidectomy, ICA mobilization, and posterior clinoidectomy. A clip was applied to the lowest accessible point of the basilar trunk at each stage. Measurements obtained during each stage included: 1). the transverse carotid oculomotor distance, that is, anteriorly between the oculomotor foramen and ICA, and posteriorly between the oculomotor nerve and ICA; and 2). the exposed length of the basilar artery, as seen under the microscope and on angiograms. RESULTS: Measurements were obtained before and after the addition of anterior clinoidectomy, mobilization of the ICA, and posterior clinoidectomy. Increases in expansion of the window and exposure of the upper basilar artery were documented as percentages of the control values. The anterior carotid-oculomotor distance averaged 7.1 mm (range, 5-10 mm) and 10.1 mm (range, 7-15 mm) before and after the additional surgical steps to expand the window, respectively. The posterior carotid-oculomotor distance averaged 12.7 mm (range, 9-18 mm) and 16.1 mm (range, 11-22 mm) before and after the additional surgical steps to expand the window, respectively. The exposed length of the basilar artery from the bifurcation to the clip was 4.2 mm (range, 1-13 mm) before expansion and 7 mm (range, 3-15 mm) after expansion. CONCLUSION: Anterior clinoidectomy and ICA mobilization increased the carotid-oculomotor space 44% anteriorly and 28% posteriorly. Posterior clinoidectomy increased the exposed length of the basilar artery by 69%. Superficial wide field exposure, expansion of the carotid-oculomotor window, and increased exposure of the upper basilar artery facilitate both visualization of the aneurysm for clip application and the use of proximal vascular control as an adjunct to basilar aneurysm surgery. PMID- 15113475 TI - Sports and pediatric cerebrospinal fluid shunts: who can play? AB - OBJECTIVE: The purpose of this study was to collect and review available data on the incidence of sport-related complications in children with cerebrospinal fluid (CSF) shunts. METHODS: After review of medical and legal literature revealed no reports on sports and shunts, an Internet-based survey was conducted of pediatric neurosurgeons. Observed complications, recommendations for sport participation, and shunt volume data were collected. RESULTS: Ninety-two providers (55% sample) responded. Seventy-seven percent had never observed a sport-related shunt complication in their practice. The remaining 23% witnessed an estimated total of 25 to 30 complications. Broken shunt catheters and shunt dysfunction were the most common complications observed. Only one acute intracranial hematoma was reported in the entire series. The incidence of sport-related CSF complications in children seems to be significantly less than 1%. Ninety percent of pediatric neurosurgeons do not restrict their patients' participation in noncontact sports. For contact sports, approximately one-third of neurosurgeons preclude all participation, whereas another third restrict specific sports. The final third of neurosurgeons surveyed do not restrict participation in contact sports for children with CSF shunts. CONCLUSION: Sport-related complications in children with CSF shunts are very uncommon. PMID- 15113476 TI - Altered arterial homeostasis and cerebral aneurysms: a review of the literature and justification for a search of molecular biomarkers. AB - DESPITE THE CATASTROPHIC consequence of ruptured intracranial aneurysms, very little is understood regarding their pathogenesis, and there are no reliable predictive markers for identifying at-risk individuals. Given that intracranial aneurysms have a strong but complex genetic component and well-characterized modifiable risk factors, it seems likely that the most valuable approach to developing minimally invasive diagnostic and prognostic tools will involve a multifactorial model that includes both genetic and environmental risk factors. Unfortunately, the genetic basis of intracranial aneurysms is poorly described, and reports describing the association of nonrandom deoxyribonucleic acid sequence variation with intracranial aneurysms have been limited to a handful of ad hoc studies that have focused on a variety of markers in small populations. One reason for this lack of coordinated analysis of the genetic basis of intracranial aneurysms is that the molecular pathogenesis and pathobiological characteristics of the disease are poorly described, so candidate marker selection has been problematic. Few studies have addressed the molecular pathological basis of intracranial aneurysms or the possible mechanisms of intracranial aneurysm formation. In this regard, candidate gene selection strategies have relied almost exclusively on limited knowledge of monogenic disorders such as Ehlers-Danlos syndrome and Marfan's syndrome, in which intracranial aneurysm is a feature of a spectrum of syndromic phenotypes. Without exception, these approaches have not affected the clinical identification and/or management of intracranial aneurysms significantly. Therefore, it is imperative that coordinated large-scale efforts in genetics, molecular biology, and genetic epidemiology are implemented to overcome these obstacles and drive developments in the field. In this review, we summarize the current screening modalities for intracranial aneurysms, review the current state of understanding relating to the genetic basis of intracranial aneurysms, and suggest a broader theory of aneurysm pathogenesis to form the foundation of a coordinated molecular search for biological markers that may be associated with aneurysm formation and rupture. PMID- 15113477 TI - Inhibition of glioma angiogenesis and invasion by SI-27, an anti-matrix metalloproteinase agent in a rat brain tumor model. AB - OBJECTIVE: The matrix metalloproteinase (MMP) inhibitor SI-27 has undergone extensive development because of its effectiveness against glioma invasion and angiogenesis. However, previous studies have been performed in vitro. The present work investigates the potential of SI-27 to inhibit tumor invasion, slow angiogenesis, and prolong survival in rodent brain tumor models. METHODS: Stable enhanced green fluorescent protein-expressing clones of a human malignant glioma cell line, U251MG, were stereotactically xenografted into the periphery of the anterior striatum and corpus callosum of Fischer 944 rats after immunosuppression with cyclosporin A. SI-27 (1 or 10 mg/kg) or carrier solution was administered on three successive days by intraperitoneal injection, and tumor invasion and angiogenesis were assessed 3 weeks later by quantitative image analysis. This was performed on whole brain sections analyzed either by direct observation of enhanced green fluorescent protein-expressing glioma cells or by additional immunohistochemistry to detect the endothelial cells with anti-factor VIII monoclonal antibody. In situ zymography on frozen sections was used to detect MMP activity. RESULTS: The group receiving a total of 30 mg/kg showed a statistically significant (P < 0.001) increase in survival time compared with the controls receiving carrier (median survival, 47.3 versus 32.6 d). There was also a decrease in MMP activity, tumor cell invasion, and neovascularization. In contrast, animals given 3 mg/kg did not show these differences. CONCLUSION: Systemic administration of the anti-MMP agent SI-27 is effective in the treatment of glioma in an animal model. PMID- 15113478 TI - Perfluorocarbon emulsion improves cerebral oxygenation and mitochondrial function after fluid percussion brain injury in rats. AB - OBJECTIVE: Cerebral ischemia is a common secondary sequela of traumatic brain injury (TBI). Experimental models of stroke have demonstrated reductions in ischemia after perfluorocarbon (PFC) administration; however, there are no published reports of PFC efficacy after TBI. The current study analyzed the effect of the PFC emulsion Oxygent (AF0144; Alliance Pharmaceutical Corp., San Diego, CA) on cerebral oxygenation, mitochondrial redox potential, and free radical formation after lateral fluid percussion injury. METHODS: After fluid percussion injury, five 2.25 ml/kg doses of PFC or saline were administered to rats breathing 100% O(2), and oxygen tension was recorded. In a second experiment, a single bolus (11.25 ml/kg) of PFC or saline was given after injury, and redox potential and free radical formation were measured at 1 or 4 hours with Alamar blue dye and dihydrorhodamine 123, respectively. RESULTS: Cerebral oxygen tension was significantly increased in both injured and sham animals treated with 11.25 ml/kg of PFC as compared with saline (P < 0.05). Likewise, PFC significantly increased mitochondrial redox potential as compared with saline at 4 hours after injury (P < 0.01). Mitochondrial peroxynitrite and peroxide production also increased with the administration of PFC (P < 0.05). CONCLUSION: The current study demonstrates that a PFC emulsion can significantly increase cerebral oxygenation after TBI and enhance mitochondrial function at 4 hours after injury as compared with saline. This study demonstrates a new therapeutic potential for PFC to enhance cerebral oxygenation and aerobic metabolism after TBI. However, the increased free radical formation with high-dose PFCs suggests the need for further studies combining PFCs with free radical scavengers. PMID- 15113479 TI - Astroglia induce cytotoxic effects on brain tumors via a nitric oxide-dependent pathway both in vitro and in vivo. AB - OBJECTIVE: In the central nervous system, astroglia produce nitric oxide (NO) in response to cytokines. We investigated whether cytokine stimulation of astroglia could inhibit brain tumor cell growth in vitro and prolong survival in vivo via an NO-dependent pathway. METHODS: Astroglia cultures were stimulated with the cytokines lipopolysaccharide and interferon-gamma and subsequently seeded with tumor cell lines. Wild-type mice and inducible NO synthase-knockout mice received in vivo cytokine stimulation followed by B16F10 murine melanoma challenge. RESULTS: Our in vitro studies demonstrate that astroglia stimulated to produce NO by the addition of cytokines dose-dependently inhibit the growth of one primary rat brain tumor cell line (9L) and three primary human brain tumor cell lines (H80, U87, and U373). This inhibition of tumor cell growth is also observed in metastatic cell lines (B16F10 melanoma, Lewis lung carcinoma, and CT26 colon). Cultured astrocytes from inducible NO synthase-knockout mice, which are incapable of induction of NO, are without the enhanced tumoricidal effect. Furthermore, when C57BL/6 mice are primed to produce NO through stereotactic intracranial administration of lipopolysaccharide plus interferon-gamma and subsequently challenged with B16F10 murine melanoma, survival is significantly prolonged, with a median survival of 26 days versus 16 days in the control group (P < 0.001). The addition of an NO synthase inhibitor (N(G)-nitro-l-arginine methyl ester) decreases this beneficial effect (median survival, 21 d). CONCLUSION: These findings suggest that NO may have an important role as a defense mechanism molecule against brain tumors; stimulation or modification of this mechanism may represent a new approach to the treatment of primary and metastatic brain tumors. PMID- 15113480 TI - Surgical construction of an in vivo carotid siphon model to test neurovascular devices. AB - OBJECTIVE: We developed an animal model to evaluate vascular trauma induced by endovascular devices that are proposed for the treatment of cerebrovascular diseases. METHODS: The model was constructed in six domestic swine by elongating the common carotid artery using interpositional grafts. Balloon catheters (n = 3), stents (n = 2), and a snare were tested 9 to 13 days after surgery. Device performance was evaluated by angiography, macroscopic photography, and histopathological examination. RESULTS: Animals tolerated the surgical procedure well, and artificial siphons were thought to provide realistic conditions for device testing. Balloon catheters induced minimal trauma, whereas coronary stents caused severe spasm or thrombosis and extensive macroscopic changes. CONCLUSION: Construction of an in vivo siphon model is feasible and potentially useful for testing neurovascular devices. PMID- 15113481 TI - Stereotactic surgery at McGill: the early years. PMID- 15113482 TI - Therapeutic basilar artery occlusion for management of medically refractory basilar artery stenosis: case report. AB - OBJECTIVE AND IMPORTANCE: To describe a novel therapeutic approach (endovascular basilar artery occlusion) to a notoriously difficult-to-manage clinical condition (actively symptomatic high-grade basilar artery stenosis) on the basis of assessment of the patient-specific mechanism of disease. CLINICAL PRESENTATION: An 81-year-old woman presented with recurrent episodes of brainstem ischemia refractory to aggressive medical therapy. Cerebral angiography revealed a high grade proximal basilar artery stenosis. On the basis of clinical presentation and angiographic findings, the pathogenesis of this complex of symptoms was thought to be embolic rather than hemodynamic. INTERVENTION: Endovascular coil occlusion of the basilar artery was used, with excellent outcome (cessation of ischemic symptoms and independent level of functioning at 1 yr). CONCLUSION: Successful endovascular management of intracranial occlusive disease requires understanding of the mechanism responsible for the patient's symptoms. PMID- 15113483 TI - Delayed cerebral hyperperfusion syndrome caused by prolonged impairment of cerebrovascular autoregulation after carotid endarterectomy: case report. AB - OBJECTIVE AND IMPORTANCE: Cerebral hyperperfusion syndrome is a rare but potentially devastating complication that typically occurs within several days after carotid endarterectomy. CLINICAL PRESENTATION: A 66-year-old man experienced asymptomatic cerebral hyperperfusion as demonstrated by single-photon emission computed tomography (SPECT) during a 2-week period after undergoing right carotid endarterectomy. This phenomenon occurred despite intensive pharmacological control of blood pressure. On the 28th postoperative day, repeat SPECT demonstrated resolution of hyperperfusion, and intensive blood pressure control was discontinued. INTERVENTION: Twelve hours later, the patient experienced left motor seizures with secondary generation. SPECT performed 36 hours after the onset of seizures demonstrated the reappearance of hyperperfusion. Intensive blood pressure control was reinstituted and maintained until the 36th postoperative day. On the next day, SPECT demonstrated resolution of hyperperfusion. CONCLUSION: The present case suggests that cerebral hyperperfusion syndrome may occur at later time points (e.g., 1 mo) after carotid endarterectomy. This delayed hyperperfusion syndrome may be related to prolonged impairment of cerebrovascular autoregulation. PMID- 15113484 TI - Parasagittal cranial fasciitis after irradiation of a cerebellar medulloblastoma: case report. AB - OBJECTIVE AND IMPORTANCE: Cranial fasciitis is a very rare, nontumoral lesion of the cranium with potential intracranial expansion typical of childhood. Radiotherapy has not been reported among the possible causes or factors associated with this condition. We present a case of cranial fasciitis in an 11 year-old patient previously admitted for cranial radiotherapy of a cerebellar medulloblastoma. Cytogenetic analysis revealed a pattern of chromosomal abnormalities suggestive of a radiation-induced lesion. CLINICAL PRESENTATION: An 11-year-old patient, who had been treated previously with craniospinal radiotherapy for a medulloblastoma, presented with a tumor resembling a parasagittal meningioma. INTERVENTION: At surgery, the tumor apparently had eroded the cranium and was deemed to originate from the external layer of the sagittal sinus. A distinct line of cleavage permitted total removal. Histological analysis was suggestive of cranial fasciitis. Cytogenetic analysis revealed the presence of a polyclonal karyotype in a background of nonclonal changes. CONCLUSION: Cranial fasciitis should be included in the differential diagnosis of intracranial tumors infiltrating the cranium. Treatment of these lesions is easier than that of other parasagittal lesions because the sinus is compressed but not infiltrated. This case is associated with previous radiotherapy; thus, cranial fasciitis could be considered one of the more common radiation-induced lesions. PMID- 15113485 TI - Selective posterior rhizotomy for painful spasticity in the lower limbs of hemiplegic patients after stroke: report of two cases. AB - OBJECTIVE AND IMPORTANCE: Selective posterior rhizotomy (SPR) has been performed mainly in children with cerebral palsy. Seldom has the use of SPR been reported for reduction of spasticity after stroke. We describe two elderly patients with hemiplegia who underwent unilateral SPR for pain caused by spasticity after stroke. CLINICAL PRESENTATION: The first patient was a 68-year-old woman who experienced spasticity and pain in her right leg during the chronic stage of a left cerebral infarction. The second patient was an 89-year-old man who had intolerable spastic pain in his left hemiplegic leg 3 months after a right cerebral infarction. INTERVENTION: Both patients underwent unilateral SPR on the spastic side to reduce the pain. After surgery, the patients' pain resolved. In the first patient, the ability to perform activities of daily living also improved. CONCLUSION: Antispastic medications are often sufficient for treatment of post-stroke spasticity. In selected cases, however, SPR can be beneficial for improving painful spasticity. PMID- 15113487 TI - Can suicide in young men be prevented by improving access and delivery among primary care services? PMID- 15113488 TI - Interprofessional collaboration and interprofessional education. PMID- 15113489 TI - Screening for colorectal cancer: decisions in general practice. PMID- 15113490 TI - Factors influencing help seeking in mentally distressed young adults: a cross sectional survey. AB - BACKGROUND: Young adults, especially men, are among those least likely to consult healthcare professionals when mentally distressed or suicidal. AIMS: To investigate the help-seeking behaviours of mentally distressed young adults. DESIGN OF STUDY: Cross-sectional survey. SETTING: Bristol and surrounding areas, including inner-city, suburban and urban locations. METHOD: A questionnaire was sent to a sample of 3004 young adults aged 16-24 years. This assessed probable mental disorder (using the 12-item general health questionnaire [GHQ-12]), suicidal thoughts (GHQ-28 suicide subscale), and help-seeking behaviours. RESULTS: Most responders who were assessed as having probable mental disorders (GHQ "cases") had not sought help. Help seeking was more common in female GHQ cases than male cases (34.8% and 21.8%,respectively; P = 0.003) and women with suicidal thoughts more commonly sought help than men with suicidal thoughts (41.6% and 30.9%, respectively; P = 0.15). Small proportions of male and female GHQ cases (7.5% and 8.9%, respectively; P = 0.6), and less than one in five responders with suicidal thoughts, had consulted a general practitioner. In more female than male cases, help was sought from family and friends (30.7% and 18.4%, respectively; P = 0.004). GHQ score was the strongest predictor of help seeking. Men had a higher threshold of severity at which they would seek help than women. Recent experience of suicidal thoughts appeared to be a stronger predictor of formal help seeking in mentally distressed women than mentally distressed men. CONCLUSION: Distressed young adults are reluctant to seek help. Men are particularly unlikely to do so unless severely distressed and tend not to seek lay support. Sex differences in help seeking may be important in understanding the high suicide rate for men. PMID- 15113491 TI - The role of primary care in the prevention of suicide and accidental deaths among young men: an epidemiological study. AB - BACKGROUND: Despite generally falling mortality and suicide rates, among young men the rates of violent death from accidents and suicide continue to rise. Most research has focused upon suicide, and the potential for effective interventions, particularly in primary care, remains controversial. AIMS: To compare health service contacts prior to suicidal and accidental deaths among young men. DESIGN OF STUDY: Examination of inquest data, postmortem and toxicology reports, and general practitioner (GP) and hospital records. SETTING: All sudden, unexpected, violent or unnatural deaths involving young men aged between 15 and 39 years and reported to the four coroner's offices of Merseyside and Cheshire during 1995. METHOD: We compared data on the timing and nature of final GP contacts before death among young men with a verdict of accident or misadventure and suicide or undetermined death. RESULTS: Out of a total of 268 violent deaths, 130 received verdicts of accident/misadventure and 97 received verdicts of suicide/undetermined death. Information on the final contact with a GP was available for 172 deaths. Although there was a significant difference between the proportion of suicide cases (56%) and that of cases of accidental death (41%) who had seen their GP during the 3 months before death, this was not significant at 1 month (38% versus 30%, respectively). Suicide cases were more likely to have seen a mental health professional at some time (27% for suicides versus 13% for accidental deaths). CONCLUSION: These findings confirm that relatively few young men consult their GP during the period before death from suicide or accidents. Prevention strategies must extend beyond suicide risk assessment, and consider ways to encourage young men to consult GPs when they are experiencing emotional distress or problems related to mental health or substance misuse. PMID- 15113492 TI - Exploratory cluster randomised controlled trial of shared care development for long-term mental illness. AB - BACKGROUND: Primary care clinicians have a considerable amount of contact with patients suffering from long-term mental illness. The United Kingdom's National Health Service now requires general practices to contribute more systematically to care for this group of patients. AIMS: To determine the effects of Mental Health Link, a facilitation-based quality improvement programme designed to improve communication between the teams and systems of care within general practice. DESIGN OF STUDY: Exploratory cluster randomised controlled trial. SETTING: Twenty-three urban general practices and associated community mental health teams. METHOD: Practices were randomised to service development as usual or to the Mental Health Link programme. Questionnaires and an audit of notes assessed 335 patients' satisfaction, unmet need, mental health status, processes of mental and physical care, and general practitioners' satisfaction with services and beliefs about service development. Service use and intervention costs were also measured. RESULTS: There were no significant differences in patients' perception of their unmet need, satisfaction or general health. Intervention patients had fewer psychiatric relapses than control patients (mean = 0.39 versus 0.71, respectively, P = 0.02) but there were no differences in documented processes of care. Intervention practitioners were more satisfied and services improved significantly for intervention practices. There was an additional mean direct cost of pound 63 per patient with long-term mental illness for the intervention compared with the control. CONCLUSION: Significant differences were seen in relapse rates and practitioner satisfaction. Improvements in service development did not translate into documented improvements in care. This could be explained by the intervention working via the improvements in informal shared care developed through better link working. This type of facilitated intervention tailored to context has the potential to improve care and interface working. PMID- 15113493 TI - Improving the ascertainment of families at high risk of colorectal cancer: a prospective GP register study. AB - BACKGROUND: Screening for colorectal cancer is effective in family members with a high risk of this condition, owing to single gene mutations. However, it is not known which is the most effective method of ascertaining these families at high risk. AIMS: To investigate whether a case-finding approach using computerised general practitioner (GP) registers would improve the ascertainment of families at high risk of colorectal cancer due to family history. DESIGN OF STUDY: Prospective GP register study. SETTING: General practices in Oxfordshire. METHOD: Identification of patients with colorectal cancer using GP registers, followed by a family history questionnaire survey to identify those at high risk. RESULTS: Using GP registers, 758 patients with a diagnosis of colorectal cancer were identified; a prevalence of 172 cases per 100 000 (95% confidence interval = 159 to 184). Of these, 305 patients, diagnosed under the age of 65 years, were sent a family history questionnaire. Two hundred and one (66%) patients responded to the survey; 10 (5%) patients were assessed as having high-risk families and 47 (23%) patients were assessed as having families at moderate risk. Eight of the high risk patients had 34 first degree relatives who would benefit from routine disease surveillance, and 37 moderate-risk patients had 153 first degree relatives. Only two high-risk and six moderate-risk patients identified were previously known to the local Clinical Genetics Department. CONCLUSION: A case finding approach using GP records and a family history questionnaire is an effective way of identifying families at high risk of developing colorectal cancer, who can then be offered disease surveillance. PMID- 15113494 TI - A primary care intervention programme for obesity and coronary heart disease risk factor reduction. AB - BACKGROUND: Obesity is a growing problem, with its associated morbidity, mortality, and economic costs. Treatment options and the availability of resources are limited and inconsistent. AIM: To implement and evaluate a primary care dietitian-run weight management programme. DESIGN OF STUDY: Pilot intervention study. SETTING: Three health centres in the north locality of Nottingham City Primary Care Trust. METHOD: Two hundred and sixteen individuals, with a body mass index (BMI) > 30 kg/m(2) and coronary heart disease risk factors, were recruited to attend education and support groups. Changes in BMI, waist circumference, percentage body fat, blood pressure, blood lipids, glycated haemoglobin (HbA(1c)), and assessment of psychological wellbeing using the "short form" (SF-36) general health questionnaire, were conducted at 0, 3, and 12 months. RESULTS: One hundred and thirty patients completed the 3-month phase, and 75 completed the follow-up 9-month phase. Four per cent of patients entering the programme achieved a 10% weight loss, and 13% achieved a weight loss between 5 and 10%. Those continuing to attend achieved a mean weight loss of 2.9% (mean = 3.1 kg, ranging from a loss of 23.6 kg to a gain of 3.8 kg, P < 0.001) at 3 months, which was maintained at 12 months. Waist circumference, percentage body fat, systolic blood pressure, total cholesterol, HbA(1c) (in those with diabetes) (P < 0.001), and triglycerides (P = 0.004) showed reduction. Psychological wellbeing improved in seven of the nine categories of the SF-36. CONCLUSION: Those who continued to attend the programme showed significant reduction in weight and other clinical parameters at 3 months, and this was maintained at 1 year with less intensive support. An attrition rate of approximately 66% by 12 months demonstrated that, in spite of intensive dietetic resources, patient retention and follow-up of progress was difficult. PMID- 15113495 TI - Access to health care prior to suicide: findings from a psychological autopsy study. AB - Previous research has suggested that progress towards suicide reduction targets may be achieved by improving the ability of general practitioners to recognise and treat mental illness. Using data from a study of suicide completers who were not in contact with specialist mental health services, we found that the rate of detection and treatment of mental health problems in primary care was high. The major barrier to receipt of care for mental health problems prior to suicide was non-consultation. The study also shows that detection and management in primary care does not necessarily result in prevention of suicide. Implications for public education, access to primary care services and the potential for suicide prevention are considered. PMID- 15113496 TI - Suicide and attempted suicide in France: results of a general practice sentinel network, 1999-2001. AB - The continuous surveillance of suicide and attempted suicide cases was added to the tasks of the French Sentinel Network of General Practitioners (GPs) in 1999. In 2001, 9700 suicides were estimated to have occurred and an estimated 61 500 attempted suicide cases were diagnosed by GPs, representing approximately 40% of cases nationwide. The majority of suicide and attempted suicide cases involved women (67%) and 43% of all cases involved patients aged 25-44 years. The fatality rate increased with age. About 80% of GPs complied with the current recommendation to refer patients who had attempted suicide to hospital. PMID- 15113497 TI - Integrated primary mental health care: threat or opportunity in the new NHS? AB - In this paper, we argue that mental illness touches everyone's lives, and that mental health care is a core activity of primary care. The increasing move towards a primary care-led National Health Service has now created a climate where primary care can move beyond providing a gatekeeper function for secondary care specialist services. Primary care is also sufficiently mature as a discipline to commission, develop, and deliver integrated patient-focused mental health services grounded in the culture and built on the strengths of primary care. We discuss examples of integrated approaches to mental health care, and highlight the potential tensions created by new ways of working. We also suggest that any changes need to be accompanied by carefully negotiated adjustments to the way primary and secondary healthcare professionals conceptualise their roles and responsibilities, and must be underpinned by new ways of learning together. PMID- 15113498 TI - New concepts in screening. PMID- 15113499 TI - Complications defining "complications". PMID- 15113500 TI - These rules are not ready yet. PMID- 15113501 TI - Influence of NICE guidelines. PMID- 15113502 TI - Impact of the Women's Health Initiative study. PMID- 15113503 TI - Post-traumatic stress disorder and primary care. PMID- 15113504 TI - Supplementary list inaccuracy. PMID- 15113505 TI - Value in venlafaxine study. PMID- 15113506 TI - More information on complementary medicine. PMID- 15113507 TI - Council tax banding and missed appointments. PMID- 15113508 TI - The GP contract: reasons to be cheerful? PMID- 15113512 TI - Does advanced access work for patients and practices? PMID- 15113513 TI - The work of the National Patient Safety Agency to improve medication safety. PMID- 15113514 TI - Evaluation of advanced access in the national primary care collaborative. AB - BACKGROUND: An aim of the National Primary Care Collaborative is to improve quality and access for patients in primary care using principles of Advanced Access. AIMS: To determine whether Advanced Access led to improved availability of appointments with general practitioners (GPs) and to examine GPs' views of the process. DESIGN: Observational study. SETTING: Four hundred and sixty-two general practices in England participating in four waves of the collaborative during 2000 and 2001. METHOD: Regression analysis of the collaborative's monthly data on the availability of GP appointments for the 352 practices in waves 1-3, and a postal survey of lead GPs in all four waves. The main outcome measures were the change in mean time to the third available appointment with GPs, and the proportion of GPs thinking it worthwhile participating in the collaborative. RESULTS: The time to the third available appointment improved from a mean of 3.6 to 1.9 days, difference = 1.7 days, 95% confidence interval (CI)= 1.4 to 2.0 days. It improved in two-thirds of practices (66% [219/331]), remained the same in 16% (53/331), and worsened in 18% (59/331). The majority of GPs in all four waves, 83% (308/371, 95% CI = 79 to 87), felt that it was worthwhile participating in the collaborative, although one in 12 practices would not recommend it. One-fifth of GPs cited a lack of resources as a constraint, and some expressed concerns about the trade-off between immediate access and continuity of care. CONCLUSION: Advanced Access helped practices to improve availability of GP appointments, and was well received by the majority of practices. PMID- 15113515 TI - Are NHS primary care performance indicator scores acceptable as markers of general practitioner quality? AB - BACKGROUND: In 2002 the Department of Health published a list of 20 indicators to judge the performance of the 302 primary care organisations (PCOs) in England during 2001-2002. General practitioners (GPs) have expressed doubts about the relevance, applicability and evidence base of these indicators for actual practice. AIMS: To fashion NHS performance indicators to be acceptable and relevant to practicing GPs. DESIGN OF STUDY: A Delphi technique followed by simple mathematical modelling. METHODS: We asked a group of 24 senior GP educators to place the Department of Health performance indicators in rank order as markers of quality in general practice. We found just seven indicators comprised 73% of the markers chosen and all seven were chosen by over three quarters of the responders. Using a simple 'sign test' system, we then calculated a composite points score for all 302 PCOs. RESULTS: We found that there were almost twice as many PCOs at the upper and lower ends of performance and fewer in the middle than we predicted theoretically. The results suggest that pan-PCO or practice factors account for the low performance scores of 16 of 35 PCOs with extremely poor performance and for the high scores of 17 of the 36 PCOs with extremely high performance. CONCLUSION: We have developed a method that shows how numerous Department of Health performance indicators can be merged into a single composite performance score. We show that this composite performance score is easy to derive, simple to interpret, is acceptable to GPs, and has face validity. PMID- 15113516 TI - Incidence and prevalence of drug-treated attention deficit disorder among boys in the UK. AB - BACKGROUND: Drug treatment for attention deficit disorder (ADD) was rare in the United Kingdom (UK) until in the mid-1990s. This contrasts with North America, where such treatment has been used to treat ADD for many decades. Since no quantitative data on the incidence and prevalence of drug-treated ADD are available in the UK, we used the general practice research database (GPRD) to obtain such information. AIMS: To provide estimates of incidence and prevalence of treated ADD in the UK for the years 1996-2001. DESIGN OF STUDY: Follow-up study of boys aged 5-14 years. SETTING: Data from UK general practices. METHODS: From the GPRD, we identified all boys aged 5-14 years who were prescribed methylphenidate for ADD. Based on the population in that age and sex category, we estimated incidence rates and the prevalence for treated ADD for the years 1996 2001. RESULTS: The incidence of first-time diagnosis of treated ADD increased among boys from the age of 5 years to reach a peak in boys aged 9-10 years, after which the incidence rate decreased. No material change in incidence was noted during the years 1996-2001. The prevalence of treated ADD was estimated to be 5.3 per 1000 boys in 1999. CONCLUSION: Drug treatment for ADD for boys treated for this disorder in the UK is substantially lower than the proportion of boys treated in North America. PMID- 15113517 TI - Detection of child mental health disorders by general practitioners. AB - BACKGROUND: Few children with mental disorders access specialist services. Although previous studies suggest that general practitioner (GP) recognition is limited, parents may not be presenting these problems. AIM: To compare GP recognition of disorders with child mental health data and to examine factors affecting recognition, in particular whether recognition is enhanced if the parent expresses concern during the consultation. DESIGN OF STUDY: A two-phase design involving an initial community survey of children between the ages of 5 and 11 years. In the second phase, primary care attenders who were regarded by their GP as having a mental health disorder were compared with those who were not. SETTING: Five general practices in Croydon, outer London. METHOD: For 186 children attending primary care, GP recognition of disorders was compared with the results of a child mental health questionnaire completed by parents. Accuracy and predictors of GP recognition were examined. RESULTS: Seventy-four per cent of children meeting criteria for caseness were not recognised by GPs as having a mental health disorder. The expression of parental concern in the consultation about a mental health problem increased the sensitivity of recognition from 26% to 88%. Expression of concern also increased GP recognition of non-cases; this reflected GP identification of other mental health and learning problems. Only a third of parents who had concerns expressed these during the consultation. CONCLUSIONS: GPs are responsive to concern and take parental views into account. As well as detecting disorders, GPs are also sensitive to other psychosocial and educational problems that may present in primary care. There is a need for parental education about child mental health disorders. PMID- 15113518 TI - Diagnosing menstrual disorders: a qualitative study of the approach of primary care professionals. AB - BACKGROUND: Menstrual disorders are a common presentation in primary care. Wide variations in management as well as discordance between patient and practitioners in relation to presenting problems have been described. AIMS: To explore the model of menstrual disorders used by practitioners in practice. DESIGN OF STUDY: Semi-structured interviews with primary care practitioners. SETTING: One inner London health authority area. METHOD: Constant comparative analysis. RESULTS: Medical practitioners were critical of the guidance provided by gynaecological definitions and texts. Practitioners put more emphasis on defining normality than on defining disorder. Practitioners used a wide range of criteria to judge their patients' complaints and decide on a course of action. Female practitioners had access to personal and professional experience and used this to develop an understanding of women's complaints. Male practitioners in particular were limited by problems in discussing menstruation in detail. Because of the difficulties in assessing patient history, other non-gynaecological factors such as patient age and consulting behaviour informed practitioners' judgements. CONCLUSION: This study draws attention to practitioners' problems in using current definitions of menstrual disorders. The combination of unhelpful medical definitions, lack of standards of normality and difficulties in discussing menstruation resulted in individual practitioners making judgements in idiosyncratic ways. In the absence of a useful gynaecological model, practitioners develop individual, often subjective and gendered models to use in practice. PMID- 15113519 TI - An epidemiological survey of symptoms of menstrual loss in the community. AB - BACKGROUND: For the early detection of gynaecological malignancy, guidance based on presenting symptoms exists to aid a general practitioner (GP) in determining who to investigate or refer. The evidence for this advice is based on the prevalence of symptoms in women with gynaecological malignancy or within specialist clinics. There are no studies on the incidence of symptoms within the community. AIM: To provide an estimate of the incidence of self-reported symptoms of menstrual loss in the community population of a single general practice. DESIGN OF STUDY: A prospective population-based cohort study of women identified through a baseline postal survey and followed 6 and 12 months later. SETTING: An urban general practice with four partners and 10,000 registered patients. METHOD: A postal baseline survey was undertaken on all women aged 18-54 years on the practice age-sex register. Responders who consented to follow-up were sent further questionnaires at 6 and 12 months. All questionnaires enquired about the presence or absence of symptoms related to vaginal bleeding. Twelve-month cumulative incidence rates were calculated using responders to the baseline, 6 month and 12-month questionnaires. RESULTS: A total of 2435 questionnaires were initially sent out at baseline and 1513 (62%) women replied to all three questionnaires. The 12-month cumulative incidence of symptoms in menstruating women was: menorrhagia 25% (95% confidence interval [CI] = 22 to 29); periods heavier than usual, 21% (95% CI = 18 to 23); change in pattern of cycle, 29% (95% CI = 26 to 32); short cycle 21% (95% CI = 19 to 24); long cycle 15% (95% CI = 13 to 18); intermenstrual bleeding 17% (95% CI = 14 to 19); postcoital bleeding 6% (95% CI = 5 to 8); prolonged period 9% (95% CI = 7 to 11). CONCLUSION: The development of symptoms of menstrual loss among women in the community is common, in contrast to the rarity of gynaecological malignancy. This raises concern about the usefulness of current guidelines, based on symptoms, advising women when to consult, and for the early detection of gynaecological malignancy in the community and primary care. PMID- 15113520 TI - Problems with a 'target' approach to access in primary care: a qualitative study. AB - We report an analysis of the qualitative phase of a study of patients' and carers' views of primary care services, focusing on their experiences of access to face-to-face general practitioner (GP) consultations during the period when new access policies were being implemented. Practices interpreted the new policy in various ways; restricted interpretations, including restriction of access to telephone booking, could cause distress to patients. Patients and carers welcomed flexible interpretations of the policy that offered choice, such as a choice of GP, or of booking in advance. PMID- 15113521 TI - Open-access versus bookable appointment systems: survey of patients attending appointments with general practitioners. AB - Access to consultations with general practitioners (GPs) is an important health policy issue. One method of providing 24-hour access is through the provision of open-access surgeries. The study aimed to compare patients' perceptions of 'bookable' and 'non-bookable' (open-access) appointments. A cross-sectional survey design was used and recruited 834 patients in a general practice. There were statistically significant differences between the bookable and the non bookable appointments for the questions on 'choice of doctor', 'whether able to see the doctor in the time they needed to', and 'convenience of the appointment'. More patients with bookable appointments saw their doctor of choice. One-fifth of patients, equally distributed between the two groups, did not feel that they were seen within the time they needed to be. Almost three-fifths of patients, equally distributed between the two groups, reported that it was either 'easy' or 'very easy' to make the appointment. Greater convenience was reported by those with bookable appointments. These findings support the hypothesis that within a single practice, there is scope for a combined appointment system in which patients can self-select, with equal satisfaction, the type of appointment that they prefer, dependent upon their own preferences or needs at the time. PMID- 15113523 TI - Systematic review of recent innovations in service provision to improve access to primary care. AB - BACKGROUND: In England, there are particularly pressing problems concerning access to adequate primary care services. Consequently, innovative ways of delivering primary care have been introduced to facilitate and broaden access. AIMS: The aim of this study was to review the evidence of seven recent innovations in service provision to improve access or equity in access to primary care, by performing a systematic review of the literature. DESIGN OF STUDY: Systematic review. SETTING: Primary care in the United Kingdom (UK). METHOD: Seven electronic databases were searched and key journals were hand-searched. Unpublished and 'grey' literature were sought via the Internet and through professional contacts. Intervention studies addressing one of seven recent innovations and conducted in the UK during the last 20 years were included. Two researchers independently assessed the quality of papers. RESULTS: Thirty studies (32 papers and two reports) were identified overall. Variation in study design and outcome measures made comparisons difficult. However, there was some evidence to suggest that access is improved by changing the ways in which primary care is delivered. First-wave personal medical services pilots facilitated improvements in access to primary care in previously under-served areas and/or populations. Walk-in centres and NHS Direct have provided additional access to primary care for white middle-class patients; there is some evidence suggesting that these innovations have increased access inequalities. There is some evidence that telephone consultations with GPs or nurses can safely substitute face-to-face consultations, although it is not clear that this reduces the number of face-to face consultations over time. Nurse practitioners and community pharmacists can manage common conditions without the patient consulting a general practitioner. CONCLUSION: The evidence is insufficient to make clear recommendations regarding ways to improve access to primary care. In the future, it is important that, as new initiatives are planned, well-designed evaluations are commissioned simultaneously. PMID- 15113522 TI - Impact of the CSM advice on thioridazine on general practitioner prescribing behaviour in Leeds: time series analysis. AB - In December 2000, the Committee for Safety of Medicines (CSM) advised that thioridazine may prolong QT intervals risking arrhythmias. We investigated the impact on general practitioner prescribing of thioridazine using a time series analysis. Numbers of items and costs of antipsychotics and benzodiazepines prescribed in Leeds from May 1999 until April 2002 were collated. Post-advice, thioridazine prescriptions dropped by 810 items per month (95% confidence interval = 420 to 1200, P < 0.001) but others increased slightly in response. Costs mimicked these changes. Fresh criteria are proposed for appraising the quality of evidence needed to inform future urgent facsimile transmissions. PMID- 15113524 TI - Sexual health. AB - In 2001 the government produced the first ever National strategy for sexual health and HIV. The strategy called for a broader role for general practice in the promotion of better sexual health. Surveys undertaken in the United Kingdom (UK) in recent years suggest that more people have more sexual partners than ever before. This has been associated with a rise in sexually transmitted infections (STIs). Screening and testing for Chlamydia trachomatis have become more widespread in the UK. Risk assessment and sexual history taking are described. They need to be carried out confidentially and non-judgmentally. Confidentiality training for all staff, including a requirement to sign confidentiality statements, is recommended. Partner notification can be done in a variety of different settings including general practice. A new course for those working in primary care has been devised, aiming to equip participants with the basic knowledge, skills, and attitudes for the effective management of STIs. PMID- 15113525 TI - Administering controlled drugs in general practice. PMID- 15113526 TI - Predictions - past and present. PMID- 15113527 TI - Fat intake and diabetes. PMID- 15113528 TI - The shipman inquiry. PMID- 15113529 TI - Changes in perception of workload. PMID- 15113530 TI - GP workloads in Europe. PMID- 15113531 TI - Centres of excellence verus community critical care. PMID- 15113532 TI - Liquid chromatographic method for the determination of lidocaine and monoethylglycine xylidide in human serum containing various concentrations of bilirubin for the assessment of liver function. AB - A high-performance liquid chromatographic method is described for determination of lidocaine (2-(dietyloamino)-N-(2,6-dimetylofenylo) acetamid) and its metabolite, monoethylglycine xylidide (MEGX), in human serum containing various concentration of bilirubin. Lidocaine and its metabolite were extracted from human serum using dichloromethane. After separation of the layers and freezing at -32 degrees C, the organic layer was decanted and evaporated under a stream of nitrogen. The sample was dissolved in the mobile phase (12% acetonitrile in 15mM potassium dihydrogen orthophosphate, pH 3.0), and after separation on a Supelcosil LC-8-DB column, the analytes were measured by ultraviolet detection at 205nm. Trimethoprim (TMP) was used as the internal standard. The recovery of the examined analytes ranged from 95.7 to 97.9% for lidocaine and from 98.0 to 99.9% for MEGX. The lower limit of quantification (LLOQ) was established at 200microg/l for lidocaine and at 10microg/l for MEGX. The choice of suitable conditions for chromatographic separation of lidocaine and its metabolite MEGX allowed the elimination of the influence of endogenous bilirubin on the result of analysis. PMID- 15113533 TI - Determination of tamsulosin in dog plasma by liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry. AB - A rapid, sensitive and accurate liquid chromatographic-tandem mass spectrometric method is described for the determination of tamsulosin in dog plasma. Tamsulosin was extracted from plasma using a mixture of hexane-ethyl acetate (2:1, v/v) and separated on a C18 column interfaced with a triple quadrupole tandem mass spectrometer. The mobile phase consisting of a mixture of methanol, water and formic acid (80:20:1, v/v/v) was delivered at a flow rate of 0.5 ml/min. Atmospheric pressure chemical ionization (APCI) source was operated in positive ion mode. Selected reaction monitoring (SRM) mode using the transitions of m/z 409-->m/z 228 and m/z 256-->m/z 166.9 were used to quantify tamsulosin and the internal standard, respectively. The linearity was obtained over the concentration range of 0.1-50.0 ng/ml for tamsulosin and the lower limit of quantitation was 0.1 ng/ml. For each level of QC samples, inter- and intra-run precision was less than 5.0 and 4.0% (relative standard deviation (R.S.D.)), respectively, and accuracy was within +/-0.3% (relative error (R.E.)). This method was successfully applied to pharmacokinetic study of a tamsulosin formulation product after oral administration to beagle dogs. PMID- 15113534 TI - Liquid chromatography-negative ion electrospray tandem mass spectrometry method for the quantification of tacrolimus in human plasma and its bioanalytical applications. AB - A simple, rapid, novel and sensitive liquid chromatography-tandem mass spectrometry method was developed and validated for quantification of tacrolimus (I) in human plasma, a narrow therapeutic index, potent macrolide immunosuppressive drug. The analyte and internal standard (tamsulosin (II)) were extracted by liquid-liquid extraction with t-butylmethylether using a Glas-Col Multi-Pulse Vortexer. The chromatographic separation was performed on reverse phase Xterra ODS column with a mobile phase of 99% methanol and 1% 10mM ammonium acetate buffer. The deprotonate of analyte was quantitated in negative ionization by multiple reaction monitoring (MRM) with a mass spectrometer. The mass transitions m/z 802.5-->560.3 and m/z 407.2-->151.9 were used to measure I and II, respectively. The assay exhibited a linear dynamic range of 0.05-25ng/ml for tacrolimus in human plasma. The lower limit of quantitation was 50pg/ml with a relative standard deviation of less than 20%. Acceptable precision and accuracy were obtained for concentrations over the standard curve ranges. Run time of 2min for each sample made it possible to analyze a throughput of more than 400 human plasma samples per day. The validated method has been successfully used to analyze human plasma samples for application in comparative bioavailability studies. The tacrolimus plasma concentration profile could be obtained for pharmacokinetic study. The observed maximum plasma concentration (C(max)) of tacrolimus (5mg oral dose) is 440pg/ml, time to observed maximum plasma concentration (T(max)) is 2.5h and elimination half-life (T(1/2)) is 21h. PMID- 15113535 TI - Determination of a cyclooxygenase II inhibitor in human plasma by capillary gas chromatography with mass spectrometric detection. AB - Sensitive methods based on capillary gas chromatography (GC) with mass spectrometric (MS) detection in a selected-ion monitoring mode (SIM) for the determination of a cyclooxygenase II (COX-II) inhibitor (3-isopropoxy-4-(4 methanesulfonylphenyl)-5,5'-dimethyl-5H-furan-2-one, I) in human plasma, in two concentration ranges of 0.1-20 and 5-1000 ng/ml, are described. Following liquid liquid extraction, the residue, after evaporation of the organic phase to dryness, was reconstituted in acetonitrile (20 l) and part of the extract (1 l) was analyzed by GC/MS/SIM. The drug (I) and internal standard (II) were separated on a 25 mx0.2 mm capillary column with HP Ultra 1 (100% dimethylpolysiloxane, 0.33 m) phase and analyzed by MS/SIM monitoring ions at m/z 237 and 282 for I and II, respectively. The standard curve was linear within the lower concentration range of 0.1-20 ng/ml and the lower limit of quantification (LLOQ) in plasma was 0.1 ng/ml. Intraday coefficients of variation (CV, n=5) were 8.9, 4.2, 5.7, 3.1, 1.9, 1.9, and 4.4% at 0.1, 0.2, 0.5, 1.0, 5.0, 10, and 20 ng/ml, respectively. The standard curve was also linear within the higher concentration range of 5 1000 ng/ml and the LLOQ in plasma was 5 ng/ml. Intraday coefficients of variation (CV, n=5) were all below 9% at all concentrations within the standard curve range. The accuracy for I in human plasma was 91-112% and the recovery of I and II was greater than 70% at all concentrations within both standard curve ranges. The details of the assay methodology are presented. PMID- 15113536 TI - Sensitive and simple gas chromatographic-mass spectrometric determination for amphetamine in microdialysate and ultrafiltrate samples. AB - A gas chromatographic-mass spectrometric (GC-MS) method is described for the measurement of amphetamine (AMP) using negative chemical ionization (NCI) mode. Without prior extraction AMP was derivatized with 2,3,4,5,6-pentafluorobenzoyl chloride (PFBC) and simultaneously extracted into toluene. The toluene extract was injected directly into GC-MS equipped with a HP-1 capillary column. The method is simple and more sensitive than most of the previously published methods. The limit of quantification of amphetamine is 25pg (1.4pg on column) with a very limited sample volume (25microl). The within-day precision was from 1.7 to 5.1% and between-day precision was from 2.2 to 7.3%. The method has been used for the measurement of several thousand microdialysate and ultrafiltrate samples and proven reliable. PMID- 15113537 TI - Determination of malondialdehyde by liquid chromatography as the 2,4 dinitrophenylhydrazone derivative: a marker for oxidative stress in cell cultures of human hepatoma HepG2. AB - Malondialdehyde (MDA) is considered a presumptive biomarker for lipid peroxidation in live organisms and cultured cells. The present study adapts an accurate and reproducible method to measure MDA by high-performance liquid chromatography (HPLC) as its 2,4-dinitrophenylhydrazone derivative in human hepatoma HepG2 cells in culture. Since MDA is assumed to increase in conditions of cellular oxidative stress, two compounds that induce pharmacological oxidative stress in cell cultures, hydrogen peroxide (H(2)O(2)) and tert-butyl hydroperoxide (t-BOOH), have been used in HepG2 cells. The results report a significant increase in the content of MDA derivative after treatment with 200 and 500microM t-BOOH for 3h, while H(2)O(2) in doses up to 500microM failed to evoke a similar response, indicating a stronger lipid peroxidation of t-BOOH to HepG2 cells than H(2)O(2). Thus, MDA can be used as a reliable biomarker for cellular oxidative stress in human hepatoma HepG2. PMID- 15113538 TI - Determination of bisphenol A in river water and body fluid samples by stir bar sorptive extraction with in situ derivatization and thermal desorption-gas chromatography-mass spectrometry. AB - A new method, based on stir bar sorptive extraction (SBSE) with in situ derivatization and thermal desorption (TD)-gas chromatography-mass spectrometry (GC-MS) is described for the determination of trace amounts of bisphenol A (BPA) in river water, urine, plasma, and saliva samples. The derivatization conditions with acetic acid anhydride and the SBSE conditions such as sample volumes and extraction time are investigated. Then, the stir bar is subjected to TD followed by GC-MS. The detection limits of BPA in river water, urine, plasma, and saliva samples are 1-5, 20, 100, and 20pgml(-1) (ppt), respectively. Calibration for BPA was shown to be linear with a correlation coefficient of >0.99. The average recoveries of BPA in all samples are higher than 95% (R.S.D. < 10%) with correction using an added surrogate standard, 13C12-bisphenol A. This simple, accurate, sensitive, and selective analytical method may be applicable to the determination of trace amounts of BPA in liquid samples. PMID- 15113539 TI - Automated solid phase extraction and quantitative analysis of human milk for 13 phthalate metabolites. AB - While the demonstrated benefits associated with breastfeeding are well recognized, breast milk is one possible route of exposure to environmental chemicals, including phthalates, by breastfeeding infants. Because of the potential health impact of phthalates to nursing children, determining whether phthalates are present in breast milk is important. We developed a sensitive method for measuring 13 phthalate metabolites in breast milk using automated solid phase extraction (SPE) coupled to isotope dilution-high-performance liquid chromatography (HPLC)-negative ion electrospray ionization-tandem mass spectrometry. We used D(4)-phthalate diesters to unequivocally establish the presence in human breast milk of enzymes capable of hydrolyzing the ubiquitous phthalate diesters to their respective monoesters. The analytical method involves acid-denaturation of the enzymes after collection of the milk to avoid hydrolysis of contaminant phthalate diesters introduced during sampling, storage, and analysis. The method shows good reproducibility (average coefficient of variations range between 4 and 27%) and accuracy (spiked recoveries are approximately 100%). The detection limits are in the low ng/ml range in 1ml of breast milk. We detected several phthalate metabolites in pooled human breast milk samples, suggesting that phthalates can be incorporated into breast milk and transferred to the nursing child. PMID- 15113540 TI - Quantitative determination of dexamethasone in bovine milk by liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry. AB - Dexamethasone (DXM) is a synthetic glucocorticoid that is authorized for therapeutic use in veterinary medicine. The European Community (EC) fixed a maximum residue limit (MRL) at 2ng/g for liver, 0.75ng/g for muscle and kidney tissues, and 0.3ng/ml for milk, while its use as growth-promoter is completely banned. The purpose of this study was to develop and validate a simple and reliable method to determine DXM residues in bovine milk. Milk proteins were removed by the addition of concentrated trichloroacetic acid and paper filtration. Solid-phase extraction clean-up on a C18 reversed phase column was performed to obtain an extract suitable for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Chromatographic separation of DXM and the internal standard desoximetasone, was achieved on a PLRP-S polymeric reversed phase column, using a mixture of 0.1% (v/v) acetic acid in water (mobile phase A) and acetonitrile (mobile phase B) as the mobile phases. They were identified using the MS/MS detection technique, and were subsequently quantified. The method has been validated according to the requirements of the EC at 0.15, 0.30 and 0.60ng/ml (being half the MRL, the MRL and double the MRL levels fixed by the EC). Calibration graphs were prepared in the 0.15-5ng/ml range and good linearity was achieved (r>or=0.99 and goodness of fit 0.999) was confirmed. Intra-day coefficient variations (CVs) for haloperidol, reduced haloperidol, bromperidol and reduced bromperidol were less than 2.5, 3.1, 2.4 and 2.5%, respectively. Inter-day CVs for corresponding compounds were 3.9, 5.1, 2.6 and 4.4%, respectively. Relative errors ranged from -5 to 10% and mean recoveries were 96-100%. The limit of quantification was 1.0ng/m for each compound. This method shows good specificity with respect to commonly prescribed psychotropic drugs, and it could be successfully applied for pharmacokinetic studies and therapeutic drug monitoring, particularly in patients receiving both haloperidol and bromperidol. PMID- 15113556 TI - Determination of Tirofiban in human serum by liquid chromatography-tandem mass spectrometry. AB - A liquid chromatography-tandem mass spectrometric (LC-MS-MS) method with a rapid and simple sample preparation was developed and validated for the determination of Tirofiban in biological fluids. Tirofiban in serum samples was extracted and cleaned up by using an automated solid phase extraction method. An external calibration was used. The mass spectrometer was operated in the multiple reaction monitoring mode (MRM). A good linear response over the range of 2-200ng/ml was demonstrated. The accuracy for Tirofiban ranged from 94.8 to 110.8% within-day and from 103.0 to 104.7% between-day. The lower limit of quantification was 2ng/ml. This method is suitable for pharmacokinetic studies. PMID- 15113557 TI - Fluorescence resonance energy transfer (FRET) using ssDNA binding fluorescent dye. AB - There is a need for simple and inexpensive methods for genotyping single nucleotide polymorphisms (SNPs) and short insertion/deletion variations (InDels). In this work, I demonstrate that a single-stranded DNA (ssDNA) binding dye can be used as a donor fluorophore for fluorescence resonance energy transfer (FRET). The method presented is a homogenous assay in which detection is based on the FRET from the fluorescence of the ssDNA dye bound to the unmodified detection primer to the fluorescent nucleotide analog incorporated into this detection primer during cyclic template directed primer extension reaction. Collection of the FRET emission spectrum with a scanning fluorescence spectrophotometer allows powerful data analysis. The fluorescence emission signal is modified by the optical properties of the assay vessel. This seems to be a completely neglected parameter. By proper selection of the optical properties of the assay plate one can improve the detection of the fluorescence emission signal. PMID- 15113558 TI - Cooperative strand invasion of supercoiled plasmid DNA by mixed linear PNA and PNA-peptide chimeras. AB - Peptide nucleic acid (PNA) is a DNA analog with broad biotechnical applications, and possibly also treatment applications. Its suggested uses include that of a specific anchor sequence for biologically active peptides to plasmids in a sequence-specific manner. Such complexes, referred to as Bioplex, have already been used to enhance non-viral gene transfer in vitro. To investigate how hybridization of PNAs to supercoiled plasmids would be affected by the binding of multiple PNA-peptides to the same strand of DNA, we have developed a method of quantifying the specific binding of PNA using a PNA labeled with a derivative of the fluorophore thiazole orange (TO). Cooperative effects were found at a distance of up to three bases. With a peptide present at the end of one of the PNAs, steric hindrance occurred, reducing the increase in binding rate when the distance between the two sites was less than two bases. In addition, we found increased binding kinetics when two PNAs binding to overlapping sites on opposite DNA strands were used, without the use of chemically modified bases in the PNAs. PMID- 15113559 TI - Engineering the pH-dependence of kinetic parameters of maize glutathione S transferase I by site-directed mutagenesis. AB - The optimisation of enzymes for particular application or conditions remains an important target in all protein engineering endeavours. Here, we report a successful strategy for altering the pH-profile of kinetic parameters and to define in detail the molecular mechanism of maize glutathione S-transferase I (GST I). To accomplish this, selected residues from the glutathione binding site (His40, Ser11, Lys41, Asn49, Gln53 and Ser67) were mutated to Ala, and the pH dependence of the catalytic parameters V(max), and V(max)/K(GSH)(m) of the mutated forms were analysed. The pH-dependence of V(max) for the wild-type enzyme exhibits two transitions in the acidic pH range with pK(a1) of 5.7 and pK(a2) of 6.6. Based on thermodynamic data, site-directed mutagenesis and UV deference spectroscopy, it was concluded that pK(a1) corresponds to GSH carboxylates, whereas the pK(a2) has a conformational origin of the protein. The pH-dependence of V(max)/K(GSH)(m) for the wild-type enzyme exhibits a single transition with pK(a) of 6.28 which was attributed to the thiol ionisation of bound GSH. These findings complement the conclusions about the catalytic mechanism deduced from the crystal structure of the enzyme and provide the basis for rationally designing engineered forms of GST I with valuable properties. PMID- 15113560 TI - Enhanced detection sensitivity using a novel solid-phase incorporated affinity fluorescent protein biosensor. AB - We engineered green fluorescent protein (GFP) into affinity fluorescent proteins (aFPs) biosensors. The aFPs detect protein-protein interactions by enhanced fluorescence intensity. In a proof of principle demonstration, aFPs containing haemagglutinin (HA) tag bind specifically to the anti-HA antibody. The sensitivity and specificity is enhanced 28-fold by incorporation of aFPs into solid-phase surface. PMID- 15113561 TI - Polystyrene surface coated with vitamin E modulates human granulocyte adhesion and MMP-9 release. AB - Vitamin E (Vit.E, alpha-tocopherol) is a natural biological antioxidant and antinflammatory agent, which protects cells from the effects of free radicals and inhibits inflammation. For such properties Vit.E has been used to improve the biocompatibility of materials such as cellulose membrane for hemodialysis. In this study granulocytes adhesion and activation have been studied after contact with normal cell culture grade polystyrene (PS) and Vit.E-coated polystyrene (Vit.E 0.1 and 0.3% (v/v)) using optical microscopy, flow cytometry and substrate zymography. Vit.E increased the number of adherent granulocytes both at 0.1% (11470 +/- 1064 cells/cm(2), P < 0.01) and 0.3% ( 13706 +/-818) cells/cm(2), P < 0.001) concentration compared to normal PS (5529+/-692 cells/cm(2)). The morphology of granulocytes adherent to Vit.E-PS appeared lightly altered and no differences have been observed in their respiratory burst compared to control granulocyte, while matrix metalloproteinase 9 or gelatinase B (MMP-9) release and activation were increased compared to the normal PS samples. Our data indicate that Vit.E-coated surface induced an increase in granulocytes adhesion and MMP-9 release in the absence of the typical oxidative stress, hallmark of granulocytes activation. A possible explanation of the phenomenon is that Vit.E modifies the surface protein adsorption thus increasing cell adhesion and in turn MMP-9 releasing. PMID- 15113562 TI - Engineering PQQ glucose dehydrogenase with improved substrate specificity. Site directed mutagenesis studies on the active center of PQQ glucose dehydrogenase. AB - Site-directed mutagenesis was carried out on the active site of water-soluble PQQ glucose dehydrogenase (PQQGDH-B) to improve its substrate specificity. Amino acid substitution of His168 resulted in a drastic decrease in the enzyme's catalytic activity, consistent with its putative catalytic role. Substitutions were also carried out in neighboring residues, Lys166, Asp167, and Gln169, in an attempt to alter the enzyme's substrate binding site. Lys166 and Gln169 mutants showed only minor changes in substrate specificity profiles. In sharp contrast, mutants of Asp167 showed considerably altered specificity profiles. Of the numerous Asp167 mutants characterized, Asp167Glu showed the best substrate specificity profile, while retaining most of its catalytic activity for glucose and stability. We also investigated the cumulative effect of combining the Asp167Glu substitution with the previously reported Asn452Thr mutation. Interpretation of the effect of the replacement of Asp167 to Glu on the alteration of substrate specificity in relation with the predicted 3D model of PQQGDH-B is also discussed. PMID- 15113563 TI - Elevation of seed alpha-tocopherol levels using plant-based transcription factors targeted to an endogenous locus. AB - Synthetic zinc finger transcription factors (ZFP-TFs) were designed to upregulate the expression of the endogenous Arabidopsis gamma-tocopherol methyltransferase (GMT) gene. This gene encodes the enzyme responsible for the conversion of gamma tocopherol to alpha-tocopherol, the tocopherol species with the highest vitamin E activity. Five three-finger zinc finger protein (ZFP) DNA binding domains were constructed and proven to bind tightly to 9 bp DNA sequences located in either the promoter or coding region of the GMT gene. When these ZFPs were fused to a nuclear localization signal and the maize C1 activation domain, four of the five resulting ZFP-TFs were able to upregulate the expression of the GMT gene in leaf protoplast transient assays. Seed-specific expression of these ZFP-TFs in transgenic Arabidopsis produced several lines with a heritable elevation in seed alpha-tocopherol. These results demonstrate that engineered ZFP-TFs comprised of plant-derived elements are capable of modulating the expression of endogenous genes in plants. PMID- 15113564 TI - Multivitamin production in Lactococcus lactis using metabolic engineering. AB - The dairy starter bacterium Lactococcus lactis has the potential to synthesize both folate (vitamin B11) and riboflavin (vitamin B2). By directed mutagenesis followed by selection and metabolic engineering we have modified two complicated biosynthetic pathways in L. lactis resulting in simultaneous overproduction of both folate and riboflavin: Following exposure to the riboflavin analogue roseoflavin we have isolated a spontaneous mutant of L. lactis strain NZ9000 that was changed from a riboflavin consumer into a riboflavin producer. This mutant contained a single base change in the regulatory region upstream of the riboflavin biosynthetic genes. By the constitutive overproduction of GTP cyclohydrolase I in this riboflavin-producing strain, the production of folate was increased as well. Novel foods, enriched through fermentation using these multivitamin-producing starters, could compensate the B-vitamin-deficiencies that are common even in highly developed countries and could specifically be used in dietary foods for the large fraction of the Caucasian people (10-15%) with mutations in the methylene tetrahydrofolate reductase (MTHFR). PMID- 15113565 TI - Improvements in metabolic flux analysis using carbon bond labeling experiments: bondomer balancing and Boolean function mapping. AB - The biosynthetically directed fractional (13)C labeling method for metabolic flux evaluation relies on performing a 2-D [(13)C, (1)H] NMR experiment on extracts from organisms cultured on a uniformly labeled carbon substrate. This article focuses on improvements in the interpretation of data obtained from such an experiment by employing the concept of bondomers. Bondomers take into account the natural abundance of (13)C; therefore many bondomers in a real network are zero, and can be precluded a priori--thus resulting in fewer balances. Using this method, we obtained a set of linear equations which can be solved to obtain analytical formulas for NMR-measurable quantities in terms of fluxes in glycolysis and the pentose phosphate pathways. For a specific case of this network with four degrees of freedom, a priori identifiability of the fluxes was shown possible for any set of fluxes. For a more general case with five degrees of freedom, the fluxes were shown identifiable for a representative set of fluxes. Minimal sets of measurements which best identify the fluxes are listed. Furthermore, we have delineated Boolean function mapping, a new method to iteratively simulate bondomer abundances or efficiently convert carbon skeleton rearrangement information to mapping matrices. The efficiency of this method is expected to be valuable while analyzing metabolic networks which are not completely known (such as in plant metabolism) or while implementing iterative bondomer balancing methods. PMID- 15113566 TI - Cofactor engineering of intracellular CoA/acetyl-CoA and its effect on metabolic flux redistribution in Escherichia coli. AB - Coenzyme A (CoA) and its thioester derivatives are important cofactors participating in over 100 different reactions in intermediary metabolism of microorganisms. The time profiles of intracellular CoA and acetyl-CoA levels were studied in an aerobic batch reactor. The CoA level starts at a high value and falls off gradually over the exponential and stationary growth phases, reaching negligible levels at the end of 24h. The acetyl-CoA level, on the other hand, increases initially reaching a maximum and decreases gradually reaching negligible levels after 24h. Overexpressing one of the upstream rate-controlling enzyme the pantothenate kinase with simultaneous supplementation of the precursor pantothenic acid to the culture medium increased the intracellular CoA/acetyl-CoA levels. It was found that supplementation of the precursor pantothenic acid is essential to increase CoA/acetyl-CoA levels. A 10-fold increase in CoA level was observed upon this overexpression in complex medium. Acetyl-CoA levels also increased (5-fold) but not as much as CoA, leaving much of the CoA in free unacetylated form. The increase in intracellular CoA/acetyl-CoA levels led to an increase in carbon flux to the acetate production pathway leading to formation of more acetate in complex medium, whereas no such change in metabolite redistribution was observed in minimal medium. PMID- 15113567 TI - Non-linear reduction for kinetic models of metabolic reaction networks. AB - Kinetic models of metabolic networks are essential for predicting and optimizing the transient behavior of cells in culture. However, such models are inherently high dimensional and stiff due to the large number of species and reactions involved and to kinetic rate constants of widely different orders of magnitude. In this paper we address the problem of deriving non-stiff, reduced-order non linear models of the dominant dynamics of metabolic networks with fast and slow reactions. We present a method, based on singular perturbation analysis, which allows the systematic identification of quasi-steady-state conditions for the fast reactions, and the derivation of explicit non-linear models of the slow dynamics independent of the fast reaction rate expressions. The method is successfully applied to detailed models of metabolism in human erythrocytes and Saccharomyces cerevisiae. PMID- 15113568 TI - Engineering of Saccharomyces cerevisiae for the production of L-glycerol 3 phosphate. AB - L-glycerol 3-phosphate (L-G3P) was accumulated in Saccharomyces cerevisiae by pathway engineering. Intracellular concentration of this metabolic intermediate could be increased more than 20 times compared to the wild type by overexpressing GPD1 encoding the glycerol 3-phosphate dehydrogenase in a gpp1 Delta gpp2 Delta mutant which lacks both isoenzymes of glycerol 3-phosphatase. Investigation of cellular pattern of triacylglycerols and glycerophospholipids did not reveal considerable changes due to accumulation of their precursor L-G3P. Hyperosmotic stress did not affect the L-G3P pool in the gpp1 Delta gpp2 Delta mutant overexpressing GPD1 despite an about 4-fold increase of specific GPD activity. In contrast, oxygen limitation improved intracellular L-G3P concentration by enhancing the availability of cytosolic NADH. The reduction of pyruvate decarboxylase activity by deleting PDC2 led to an additional increase. In fact, the triple mutant gpp1 Delta gpp2 Delta pdc2 Delta overexpressing GPD1 accumulated 17 mg L-G3P/g dry weight during glucose batch fermentation under oxygen limitation. This value corresponds to an about 100-fold increase compared to that found in the wild type. PMID- 15113569 TI - Effect of zwf gene knockout on the metabolism of Escherichia coli grown on glucose or acetate. AB - The mutant deficient in glucose-6-phosphate dehydrogenase (G6PDH) was constructed by disrupting zwf gene by one-step inactivation protocol using polymerase chain reaction primers. The knockout of zwf gene was shown to have different influence on the metabolism of Escherichia coli grown on glucose or acetate. The decreased rates of substrate uptake and CO(2) production were found for the mutant grown on acetate, whereas these two rates were increased during the growth on glucose. The metabolic flux analysis based on (13)C-labeling experiments indicates that the metabolism of the mutant grown on glucose is related to the higher flux via tricorboxylic acid (TCA) cycle to generate anabolic reducing equivalents normally provided by the oxidative pentose phosphate pathway. However, the metabolism of the mutant grown on acetate shows a lower flux towards the TCA cycle as compared with the parent strain. The decreased flux through TCA cycle is associated with an increased flux via the glyoxylate shunt, by which the carbon source can bypass the two decarboxylative steps of TCA cycle in which CO(2) is released, thus conserving more carbon for biosynthesis in response to the decreased uptake rate of the carbon source. PMID- 15113570 TI - Congenital maxillomandibular fusion: a case report and review of the literature. AB - Congenital fusion of mandible to maxilla (syngnathia) is rare. Only 25 cases of syngnathia have been reported up to date since the first case was reported in 1936. Of these cases only six have involved fusion of the ascending rami of mandible to the maxilla and zygomatic complex. This report is an additional case of syngnathia involving bilateral bony fusion of mandibular horizontal and ascending rami to zygomatic complex and maxilla in a female neonate. Delay in treatment afforded the opportunity to observe the effects on the growth of the patient as she presented again at the age of 3 years. PMID- 15113571 TI - Biomechanical and clinical implications of distraction osteogenesis in craniofacial surgery. AB - INTRODUCTION: Craniofacial distraction osteogenesis is an established surgical procedure to correct bony malformations. Force transduction through the osteotomized bone fragments elicits defined biological responses in the gap tissue, which determines the clinical success of the distraction treatment. OBJECTIVE: The purpose of this investigation was to evaluate clinically a new distraction protocol based on an analysis of the biological and biomechanical parameters executing direct effects on bone regeneration during distraction. STUDY DESIGN: A multistep distraction protocol was used in 39 patients and the clinical outcome was monitored postoperatively. RESULTS: All the distraction cases were successful with a single exception. Segmental displacements were stable clinically and radiologically. CONCLUSION: In order to improve the clinical success of distraction osteogenesis, individual treatment protocols are recommended. PMID- 15113572 TI - Advancement of the anterior maxilla by distraction (case report). AB - INTRODUCTION: Several techniques of distraction osteogenesis have been applied for the correction of compromised midface in patients with clefts of the lip, alveolus and palate. PURPOSE: This article presents a technique of callus distraction applied in a specific case of hypoplasia of a cleft maxilla with the sagittal advancement of the maxilla thus not affecting velopharyngeal function. CONCLUSION: The decision to apply distraction osteogenesis for advancement of the anterior maxillary segment in cleft patients offers many advantages. PMID- 15113573 TI - Condylar reconstruction by oblique sliding vertical-ramus osteotomy. AB - INTRODUCTION: The posterior border of the mandibular ramus can be used as a pedicled graft for reconstruction of the condyle. This article describes a technique that is better in certain situations than the use of other autogenous grafts or alloplastic materials. PATIENTS AND METHODS: Three adult patients, two with osteochondroma and one with hyperplasia were treated by condylectomy and simultaneous reconstruction with the pedicled posterior mandibular border. A meticulous oblique osteotomy and reinsertion of the lateral pterygoid muscle was performed. RESULTS: In all three cases an immediate mouth opening with stable occlusion was achieved. The interincisal opening was more than 40 mm after 3 weeks, with a deviation no greater than 4mm towards the affected side. All excursive movements were present in all directions, and correction of the facial asymmetry was achieved. There was no T.M.J. pain and all patients expressed satisfaction during the follow-up of 56 months (average). An adequate remodelling of the neocondyle without resorption as well as a stable occlusion was observed in every case. CONCLUSION: Reconstruction of the condyle by sliding vertical oblique ramus osteotomy provides, in cases of condylar tumours, excellent functional and cosmetic results. PMID- 15113574 TI - Odontogenic tumours, a collaborative retrospective study of 75 cases covering more than 25 years from Estonia. AB - INTRODUCTION: The aim of the present collaborative study was to analyse retrospectively the character of odontogenic tumours in Estonia, involving the entire Estonian population (1.4 million), and to compare their prevalence with the figures presented in similar reports from other countries. MATERIAL AND METHODS: All material for the retrospective study was retrieved from the files of the Departments of Maxillofacial Surgery in Tartu and Tallinn, Estonia, where all in/out-patients are treated from the whole country. The final diagnosis in each case of odontogenic tumour was based on the 1992 WHO histological criteria. RESULTS: A total of 75 odontogenic tumours was found, 74 (98.6%) of which were benign, and 1 (1.3%) was malignant. The frequency of odontogenic tumours in this study was the lowest ever reported. The most common tumours were odontoma (34.3%), followed by ameloblastoma with different subtypes (25.3%), ameloblastic fibroma (16%), odontogenic myxoma (12%) and benign cementoblastoma (8%). CONCLUSION: Odontogenic tumours are relatively rare in Estonia compared with the data from other countries. PMID- 15113575 TI - Primary intraosseous carcinoma of the jaws arising from an odontogenic cyst--a case report. AB - BACKGROUND: Squamous cell carcinoma of the jaw located purely in the bone is extremely rare. Most of these intraosseous carcinomas, also called odontogenic carcinomas are thought to arise from the epithelial lining of an odontogenic cyst. CASE REPORT: A primary intraosseous carcinoma arising from an odontogenic cyst in a 64-year-old woman is reported. No subjective symptoms were noted by the patient. Multiple retained teeth with associated presumed cystic lesions were evident in the lower jaw on a routine radiograph. Histology revealed an intraosseous carcinoma after removal of the teeth and "cystectomy". No metastasis was detected. Segmental resection of the mandible, selective neck dissection and reconstruction with an iliac bone graft was performed. The patient is free of disease after 3 years. CONCLUSION: Although primary intraosseous carcinoma is rare, this case emphasizes the importance of careful histological examination of apparently innocuous odontogenic cysts. In addition, as malignant changes in their epithelial lining are always possible, "cystic" lesions should not only be removed but as completely as possible. PMID- 15113576 TI - Eosinophilic granuloma: resolution of maxillofacial bony lesions following minimal intervention. Report of three cases and a review of the literature. AB - INTRODUCTION: Langerhans' cell histiocytosis is a collective term used to describe a group of enigmatic proliferative disorders. The natural history of the disease varies from a slow, benign, localized symptomatic bony or soft tissue lesion, to a rapidly progressive widespread multiple organ disorder which is often fatal. Eosinophilic granuloma accounts for 60-70% of all cases of Langerhans' cell histiocytosis and can present as solitary (50-75%) or multifocal defects in bone. It occasionally presents as a localized soft tissue lesion. There are multiple treatment options but the response is unpredictable. AIMS: We present three separate cases, of the maxillofacial skeleton where the lesions of eosinophilic granuloma resolved following incisional biopsy only. PATIENTS AND RESULTS: Three patients presented with solitary lesions of the maxillofacial skeleton. All were diagnosed as Langerhans' cell histiocytosis following open curettage, which also resulted in resolution of the lesions. Follow-up has thus far been disease free. CONCLUSION: For some solitary Langerhans' cell histiocytosis lesions, simple curettage is the only treatment required. The paper discusses the need to confirm the solitary nature of the disease and the need for follow-up. Reviewing the literature on the disease, the authors suggest that perhaps cellular immaturity holds the cells of the lesion in a disease state until pushed to maturity by the trauma of open curettage surgery, resulting in a complete resolution of the disease. PMID- 15113577 TI - Angiogenic factors in squamous cell carcinoma of the oral cavity: do they have prognostic relevance? AB - INTRODUCTION: Angiogenesis is a cascade-like mechanism which is essential for tumour growth and metastasis. Therefore the existence of angiogenic molecules and the density of activated endothelial cells in individual tumours is of major interest. MATERIAL/PATIENTS: In order to evaluate the prognostic significance of these molecules, the distribution pattern was studied of vascular endothelial growth factor (VEGF) and activated endothelial cells in tumours and normal, healthy oral mucosal specimens from 51 consecutive patients with primary oral squamous cell carcinoma. STUDY DESIGN: Frozen sections (vascular endothelial growth factor) and paraffin-embedded sections (endoglin, CD105) were investigated quantitatively by immunohistochemistry. The Pearson correlation, the non parametric Mann-Whitney test, the non-parametric Wilcoxon rank sum test with multiple comparisons and the non-parametric Kruskal-Wallis test with multiple comparisons were used for statistical analyses. RESULTS: Endoglin expression in tumour tissue was significantly higher than in normal healthy mucosa (P<0.001). T1 tumours showed a significantly lower staining for endoglin compared with T2, T3 and T4 tumours but there was no increase with each T stage. No statistical correlation was found between VEGF expression and endoglin staining. CONCLUSIONS: Even though there is controversy about the prognostic relevance of VEGF, our results suggest that the factor is not suitable to decide prognosis in oral cancer. Endoglin may have a significant role in the development of squamous cell carcinoma of the oral cavity and might be relatively more specific than commonly used endothelial markers. PMID- 15113578 TI - Optimisation of powders for pulmonary delivery using supercritical fluid technology. AB - Supercritical fluid technology exploited in this work afforded single-step production of respirable particles of terbutaline sulphate (TBS). Different crystal forms of TBS were produced consistently, including two polymorphs, a stoichiometric monohydrate and amorphous material as well as particles with different degrees of crystallinity, size, and morphology. Different solid-state and surface characterisation techniques were applied in conjunction with measurements of powder flow properties using AeroFlow device and aerosol performance by Andersen Cascade Impactor tests. Improved fine particle fraction (FPF) was demonstrated for some powders produced by the SCF process when compared to the micronised material. Such enhanced flow properties and dispersion correlated well with the reduced surface energy parameters demonstrated by these powders. It is shown that semi-crystalline particles exhibited lower specific surface energy leading to a better performance in the powder flow and aerosol tests than crystalline materials. This difference of the surface and bulk crystal structure for selected powder batches is explained by the mechanism of precipitation in SCF which can lead to surface conditioning of particles produced. PMID- 15113579 TI - Comparison of different mathematical models for the tensile strength-relative density profiles of binary tablets. AB - During the last decade the evolution of the pharmaceutical dosage form design has been important. In controlled release matrix tablets, the tensile strength is an essential parameter to consider, because a minimal mechanical strength is needed for tablet production, handling and avoidance of any dose dumping during its use. Recent developments in percolation theory led to the theoretical proposal of lattice strength that was applied to the tensile strength of tablets. This mechanical property was described as a power law of the relative density involving a critical value that corresponds to the percolation threshold. The objective of the present work is to estimate these mechanical thresholds in KCl Ethocel100 tablets that were manufactured from different sieve fractions (100 150, 150-200, 250-300 microm). Three power law models are compared regarding the best fit of the tensile strength-relative density profiles. The main criteria for this choice are the Akaike's Information Criterion (AIC), the analysis of the residuals in conjunction with the soundest physical meaning of the models. Accordingly, a power law model was chosen that assumes an initial strength parameter. No correlation could be established between the different mixture ratios or sieve fractions with the critical relative densities. The study showed that an equation based on percolation theory can adequately model tablet strength density profiles from matrix tablets. PMID- 15113580 TI - Stereospecific chemical and enzymatic stability of phosphoramidate triester prodrugs of d4T in vitro. AB - The phosphoramidate triester prodrug approach is widely used to deliver nucleotide forms of nucleoside analogues into target cells. We investigated the stereoselective stability of a series of prodrugs of the anti-HIV agent 2',3' didehydro-2',3'-dideoxythymidine (d4T). Chemical stability was evaluated in phosphate buffer at pH values of biological relevance (i.e. pH 2.0, 4.6, 7.4). Enzymatic stability was tested in human plasma, in Caco-2 cell homogenates and monolayers and in rat liver. The compounds were relatively stable to chemical hydrolysis. Between 50 and 70% of unchanged prodrug was recovered after 16h incubation in human plasma, with no stereoselective preference for phosphate diastereoisomers. The p-OMe phenyl derivative, however, was an exception and only 5% of one diastereoisomer was recovered. In Caco-2 cells the stability and stereoselectivity largely depended on the experimental conditions: high enzymatic activity and stereoselectivity was observed in cell homogenates, but not in monolayers. In rat liver S9 fractions the stability profile was similar to that in Caco-2 cells and carboxyl ester cleavage appeared to be the sole mechanism of degradation in both media. The large and unpredictable differences in stereoselective metabolic rate of the pronucleotide series here presented suggest that in vivo circulating levels of intact prodrug could exert profoundly different activity or toxicity due to preferential body distribution of one diastereoisomeric form. PMID- 15113581 TI - PAMPA--a drug absorption in vitro model 8. Apparent filter porosity and the unstirred water layer. AB - In the parallel artificial membrane permeability assay (PAMPA), if more lipid is used than needed to fill all the pores of a microfilter, the excess lipid layer on both sides of the lipophilic filter increases the "apparent" porosity, epsilon(a), of the filter. The specific resistance of the artificial membrane barrier is lowered with increasing lipid excess. If this effect is not recognized, and the uncorrected value of filter porosity, epsilon, is used, then the calculated intrinsic permeability and the unstirred water layer (UWL) permeability coefficient of the permeating molecule can be significantly overestimated, resulting in underestimates of the thickness of the unstirred water layer. Unstirred water layer corrections are important in pharmaceutical research for in vitro-in vivo correlations aimed at predicting oral absorption and blood-brain barrier penetration characteristics of lead candidate compounds. The novel concept of the apparent porosity is introduced, described, and its utility demonstrated with the drugs diclofenac, desipramine, caffeine, and piroxicam. The PAMPA data of Wohnsland and Faller [J. Med. Chem. 44 (2001) 923] is taken as an example, where the reported extraordinarily efficient stirring is thought to be better explained in terms of normal stirring when apparent porosity is taken into account in the calculation of the effective permeability coefficient. PMID- 15113582 TI - Design, synthesis, and computational affinity prediction of ester soft drugs as inhibitors of dihydrofolate reductase from Pneumocystis carinii. AB - A series of dihydrofolate reductase (DHFR) inhibitors, where the methylenamino bridge of non-classical inhibitors was replaced with an ester function, have been prepared as potential soft drugs intended for inhalation against Pneumocystis carinii pneumonia (PCP). Several of the new ester-based inhibitors that should serve as good substrates for the ubiquitous esterases and possibly constitute safer alternatives to metabolically stable DHFR inhibitors administered orally, were found to be potent inhibitors of P. carinii DHFR (pcDHFR). Although the objectives of the present program is to achieve a favorable toxicity profile by applying the soft drug concept, a high preference for inhibition of the fungal DHFR versus the mammalian DHFR is still desirable to suppress host toxicity at the site of administration. Compounds with a slight preference for the fungal enzyme were identified. The selection of the target compounds for synthesis was partly guided by an automated docking and scoring procedure as well as molecular dynamics simulations. The modest selectivity of the synthesized inhibitors was reasonably well predicted, although a correct ranking of the relative affinities was not successful in all cases. PMID- 15113583 TI - The application of non-contact laser profilometry to the determination of permanent structural change induced by compaction of pellets II. Pellets dried by different techniques. AB - Microcrystalline (MCC) pellets of different structural and mechanical properties were produced by the process of extrusion and spheronization from the formula MCC:water:ethanol (5:3:2) using four different drying techniques, namely: freeze drying, fluid-bed drying, hot air oven drying and desiccation with silica-gel. Six hundred milligrams of these pellets were compacted by 130 MPa to flat-faced tablets and stored for 48 h in ambient temperature and humidity after which their permanent structural change (plastic deformation) was investigated in terms of surface roughness parameters using a non-contact laser profilometer. The results were compared with the deformability values measured as a reciprocal of the slope of the force/displacement curve obtained during diametral compression test of the individual pellets. Based on the different rate of moisture removal, means of heat and mass transfer, and static and dynamic nature of the bed the different drying techniques produced pellets of different porosity, strength and deformability. The increase in deformability of the pellets with the increase of porosity was illustrated by the reduction of the surface roughness parameters. Analysis of variance identified the significant difference in the mean rugosity values of the tablets from the pellets produced by the various drying techniques. The deformability values obtained were reasonably comparable to those plasticity values explained in terms of the inverse of the slope of the force/displacement curves. The laser profilometry technique was able to quantify the permanent structural change of the pellets after compaction in terms of mean rugosity values. The methodology was able to incorporate a wide variety of deformable pellets of the same formulations but produced by different drying techniques. The porous freeze-dried pellets produced the smoothest tablet surface profile, while the other techniques increased the rugosity values in ascending order from fluid bed drying, desiccation with silica-gel to hot air oven drying technique. PMID- 15113584 TI - Bupivacaine containing dry emulsion can prolong epidural anesthetic effects in rabbits. AB - To assess the prolongation of epidural bupivacaine by a novel lipid formulation, a physically stabilized bupivacaine containing dry emulsion was prepared by spray drying. Bupivacaine release from the oil-in-water emulsion was studied using an in vitro two-phase stirred model, then the pharmacodynamic effects and the pharmacokinetics of bupivacaine from the spray-dried emulsion were evaluated and compared to a bupivacaine hydrochloride solution, following a two-period cross over epidural administration in rabbits. The in vitro release characteristics suggested an extended release of bupivacaine from the emulsion compared to the solution. From the in vivo study, C(max) obtained with the emulsion (containing 5 mg bupivacaine) was not statistically different than from the solution (containing 2 mg bupivacaine) while T(max) was increased, suggesting a diminution of bupivacaine systemic absorption. The onset time of epidural anesthesia was similar for both formulations of bupivacaine used, while a significant blockade prolongation (360%) was observed with the emulsion compared to the solution, suggesting a controlled release of bupivacaine. Dry emulsions could be promising dosage forms to optimize the disposition of epidurally administered LAs. PMID- 15113586 TI - Diagnostic principles and new developments in primary cutaneous B-cell lymphomas. AB - The most common subtypes of primary cutaneous B-cell lymphomas are marginal zone B-cell lymphoma/immunocytoma, follicle center cell lymphoma, and large B-cell lymphoma of the leg. Precise classification (European Organization for Research and Treatment of Cancer (EORTC) scheme) can be achieved only after a complete synthesis of clinical, histopathological, immunophenotypic, and molecular features. It is extremely important to emphasize that primary cutaneous B-cell lymphomas differ significantly from their nodal counterparts and are frequently characterized by an excellent prognosis. Awareness of this special clinical behavior should prevent the application of unnecessarily aggressive treatment protocols. Future definitions of primary cutaneous B-cell lymphomas will be based on their etiology and pathogenesis and especially on their molecular features. Some important areas are presented where exciting findings have been detected. PMID- 15113585 TI - Effect of probucol on the oral bioavailability of cyclosporine A. AB - We have previously reported the reduction in oral bioavailability of cyclosporine A (CsA) by probucol, a lipid-lowering drug. To evaluate the mechanism, we examined the effect of probucol on the transport of CsA across Caco-2 cell monolayers, on the influence of CsA pharmacokinetics in rats, and on the change of ultraviolet-absorption spectrum of the drug. Pretreatment with probucol (50 microM) inhibited (P < 0.001) both the apical-to-basal (-73.1%) and basal-to apical (-77.8%) fluxes of [3H]-CsA. In rats, probucol orally given 6 h after, but not simultaneous with CsA did not decrease peak CsA concentration or area under the blood CsA concentration-time curve following a single oral dosing of CsA after the pretreatment with probucol for 7 days. These data indicate that P glycoprotein-mediated active transport from intracellular to apical is not involved in the mechanism of probucol-CsA interaction, and absorption of CsA decreases in the presence of probucol in the gastrointestinal tract. In difference spectral analysis, probucol reduced the absorption peak of CsA in a concentration-dependent manner, indicating that probucol could form a complex with CsA. These results suggest that probucol interferes with CsA absorption probably by physicochemical mechanism such as complex formation, but not the enhancement of P-glycoprotein function. PMID- 15113587 TI - Prostanoids in the cutaneous immune response. AB - Prostanoids, consisting of the prostaglandins and the thromboxanes, are the cyclooxygenase metabolites of arachidonic acid. They exert a range of actions mediated by their respective receptors expressed in the target cells. In the skin, it is well known that prostanoids are abundantly produced and that the prostanoid receptors are highly expressed. However, the physiological role of prostanoids in the skin has not been clarified. Recent developments in the molecular biology of the prostanoid receptors have enabled the investigation of the physiological roles of each receptor by disruption of the respective genes in combination with prostanoid receptor selective compounds. Here, we review novel findings relating to the roles of prostanoids in the cutaneous immune responses. These may prove useful in the development of new therapeutic agents that can selectively manipulate the actions mediated by each receptor. PMID- 15113588 TI - Microarray analysis of UVB-regulated genes in keratinocytes: downregulation of angiogenesis inhibitor thrombospondin-1. AB - BACKGROUND: Ultraviolet (UV) B light is an environmental mutagen that induces changes in cutaneous gene expression leading to immune suppression and carcinogenesis. Keratinocytes are a primary target for UVB. OBJECTIVE: To further delineate UVB-induced gene expression changes in keratinocytes. METHODS: cDNA microarray technology was utilized to examine gene expression in normal human KC (NHKC) following 20 mJcm(-2) UVB irradiation. Data was confirmed by semi quantitative RT-PCR. RESULTS: Microarray analysis revealed 57 genes were upregulated, and 27 genes were downregulated, by at least two-fold following UVB. One downregulated gene was the endogenous angiogenesis inhibitor thrombospondin-1 (TSP-1). Semi-quantitative RT-PCR confirmed persistent downregulation of TSP-1 up to 18h following UVB. Microarray analysis also revealed upregulation of platelet derived endothelial cell growth factor (PD-ECGF)--an angiogenesis activator. CONCLUSION: Our results suggest a gene expression mechanism by which UVB induces an angiogenic switch in keratinocytes. This may represent an important early event promoting neovascularization and growth of cutaneous neoplasms. PMID- 15113589 TI - The G2028R glycine substitution mutation in COL7A1 leads to marked inter-familiar clinical heterogeneity in dominant dystrophic epidermolysis bullosa. AB - BACKGROUND: Glycine substitution mutations in COL7A1 not only cause dominant dystrophic epidermolysis bullosa (DDEB), but can also be silent mutations which lead to recessive dystrophic epidermolysis bullosa (RDEB) in combination with additional mutations in the other allele. OBJECTIVE: In this study, we have examined a large American Caucasian pedigree in which 10 family members from four generations presented with simple toenail dystrophy without skin fragility in autosomal dominant manner. METHOD: We sequenced COL7A1 of this pedigree. RESULTS: Mutational analysis indeed detected a heterozygous G-to-A transition at nucleotide position 6082 leading to G2028R in all the affected members. Surprisingly, mutation database revealed that this G2028R mutation had been previously identified in two distinct Asian families with DDEB showing apparent skin fragility and blister formation. One case was a 17-month-old Chinese female with classical phenotype of DDEB and the other was a 27-year-old Japanese female with typical epidermolysis bullosa (EB) pruriginosa. To better understand the molecular mechanisms of this marked inter-familiar clinical heterogeneity, we examined the entire sequence of all the exons and exon-intron borders as well as the promoter region of COL7A1 in all the three families. Sequence results demonstrated no significant nucleotide difference in COL7A1 among the three pedigrees. CONCLUSION: This paper has demonstrated for the first time that identical COL7A1 glycine substitutions can cause remarkably heterogeneous clinical phenotypes extending from simple toe nail dystrophy without skin fragility to typical DDEB and EB pruriginosa. In addition, the fact of inter familiar, not intra-familiar clinical heterogeneity associated with G2028R suggest that the other molecular mechanisms not controlled by COL7A1 coding sequence might be responsible for the clinical heterogeneity. PMID- 15113590 TI - Both Th2 and Th1 chemokines (TARC/CCL17, MDC/CCL22, and Mig/CXCL9) are elevated in sera from patients with atopic dermatitis. AB - BACKGROUND: Chemokines and their receptors are important elements for the selective attraction and activation of various subsets of leukocytes. Expression of CXCR3 ligands, such as monokine induced by IFN-gamma (Mig) leads to preferential Th1 recruitment, whereas CCR4 ligands, thymus and activation regulated chemokine (TARC) or macrophage derived chemokine (MDC), mediate preferential Th2 recruitment. Although atopic dermatitis (AD) has been shown to be a Th2-type disease, recent studies have revealed that Th1-type cytokines, such as IFN-gamma, especially in chronic skin lesions, play important roles in pathogenesis of AD. OBJECTIVE: The purpose of this study was to investigate serum levels of Th2 chemokines TARC and MDC and a Th1 chemokine Mig in the same samples from patients with AD and their clinical correlation. METHODS: Serum chemokine levels in patients with AD (n = 55), contact dermatitis (CD; n = 15), and normal controls (n = 30) were examined by ELISA. RESULTS: Serum levels of TARC and MDC in AD patients and CD patients were significantly higher than those found in normal controls. Serum levels of these chemokines were similar for AD patients and CD patients. Furthermore, these levels correlated positively with disease severity, total IgE levels, and peripheral eosinophilia in AD patients. Serum Mig levels in AD patients and CD patients were significantly higher than those in control subjects. However, serum Mig levels were significantly elevated in CD patients relative to AD patients. Furthermore, serum Mig levels correlated positively with levels of both TARC and MDC in AD patients. CONCLUSION: These results suggest that both Th2 and Th1 chemokines may play roles in the development of AD. PMID- 15113591 TI - Formalin-fixed tumor cells effectively induce antitumor immunity both in prophylactic and therapeutic conditions. AB - BACKGROUND: Autologous whole tumor cell-based vaccinations would seem to be ideal since such vaccinations, in contrast to vaccination with a single defined antigen, have the potential to elicit a broad type of T-cell immune response to tumor-associated antigens. OBJECTIVE: We modified formaldehyde (formalin)-fixed mouse melanoma cells and investigated the utility of those cells as sources of tumor antigens for immunotherapy. METHODS: C57BL/6 or the proteasome activator PA28alpha-knockout mice were intradermally inoculated with 1% formalin-fixed B16 cells three times at weekly intervals either before or after tumor challenge. Simultaneously, interleukin-12 gene was transferred into the skin around immunization sites using gene gun technology. The effects were evaluated by tumor growth, antigen-specific interferon-gamma production in splenic lymphocytes, and activation of dendritic cells. RESULTS: Fixed cells directly induced production of tumor necrosis factor-alpha in dendritic cells more effectively than did frozen and thawed cells. More than 60% of the mice immunized with fixed cells and interleukin-12 rejected the challenged B16 tumor. CD4+ T cells from those mice produced a significant amount of interferon-gamma in response to melanoma cells. Furthermore, this combined treatment showed antitumor immunity initiated by CD8+ and CD4+ T cells in the therapeutic experiments. PA28alpha/beta appeared not to be required for the development of CD8+ T cells, although it is known to be essential for the development of CD8+ T cells specific for tyrosinase-related protein-2, one of melanocyte-lineage differentiated antigens. CONCLUSION: These results suggest that formalin-fixed autologous melanoma cells have a potential to function as effective antigen sources for immunotherapy. PMID- 15113592 TI - The effect of the long-term cultivation on telomere length and morphology of cultured epidermis. AB - BACKGROUND: Cultured epidermis has been successfully used in clinical treatment such as burns and pigmentary disorders. Although the generation of wide cultured epidermis for clinical use may require repeated passages, especially for allografts, the effects of long-term cultivation on its quality and cell viability are not well known. OBJECTIVES: To investigate the changes in morphology, telomere length, and telomerase activity during the passages of cultured epidermis and keratinocytes up to the passage limit, and to examine the usefulness of telomere length as a performance criterion for cultured epidermis. METHODS: The keratinocytes obtained from five patients were used to generate cultured epidermis. At the early passage and after cultivation up to the passage limit, morphology, telomere length and telomerase activity were investigated by using microscopes, southern blot analysis and telomeric repeat amplification protocol assay, respectively. RESULTS: The cultured cells started to show morphological changes when each passage was close to its limit and the cell sheets assumed an irregular stratification with various sizes of cytoplasm and nuclei. At the passage limit, the telomere length had decreased approximately 80 85%, and the average telomerase activity had declined under serum-free culture conditions. CONCLUSION: The results of this study showed the morphological change and telomere length reduction by long-term cultivation on cultured epidermis. Although the reduction in telomere length and telomerase activity may not be the major cause of the senescence, they could provide a useful information for the quality of the cultured epidermis. PMID- 15113593 TI - The effect of carotenoids and tocopherols in the protection of human fibroblast cells against UVA-induced DNA damage. PMID- 15113594 TI - In vivo transfer of TGF-alpha and beta genes to keratinocytes. PMID- 15113595 TI - Determination of thymus and activation-regulated chemokine (TARC)-contents in scales of atopic dermatitis. AB - BACKGROUND: Thymus and activation-regulated chemokine (TARC) is a cytokine which selectively controls the migration of type 2-helper T lymphocytes into inflammatory lesions, and the serum level is strongly associated with disease severity of atopic dermatitis (AD). OBJECTIVE: To examine the role of TARC in the pathogenesis of AD, we determined TARC-contents in the scales obtained from lesional skin of the patients with AD. RESULTS: High amount of TARC was detected in the scales of lesional skin obtained from the patients with AD, and the amount was well correlated with the serum IgE levels but not with the blood eosinophil counts. The TARC-content in the lesional scales was not correlated with a-431C/T polymorphism of TARC promotor gene, suggesting other regulating mechanisms in TARC production in the lesion. CONCLUSION: High amount of TARC is produced in the kesion of AD, and analysis of cytokine content in lesional scales may provide some tools to clarify the pathogenesis of AD. PMID- 15113596 TI - A qualitative retrospective analysis of positive control data in developmental neurotoxicity studies. AB - Testing for neurodevelopmental effects commonly involves both functional and neuropathological assessments in offspring during and following maternal exposure. The use of positive controls in neurotoxicity screening has been advocated by numerous expert groups. Evaluation of positive control data allows evaluation of laboratory proficiency in detecting changes in the structure and function of the developing nervous system and comparison of the sensitivity of assessments in different studies and laboratories. This project surveyed approaches taken in contract and industrial laboratories in generating and providing these data. Positive control data submitted in support of 34 developmental neurotoxicity (DNT) studies from 16 different laboratories were summarized by test method for information on the following: age relevance of test subjects, the presence of a dose-response relationship, gender, group size, statistics, report quality, quality assurance, and the year the study was conducted. Endpoints included the following: developmental landmarks, clinical observations (CO), motor activity, startle response, learning and memory, qualitative neuropathology, and quantitative brain morphometry (linear measurements of selected brain regions). Results ranged from no positive control data for three laboratories, to one laboratory that submitted 17 separate positive control reports. The qualitative range was similarly broad, from excellent to poor. Various problems were identified, including the following: inappropriate report structure (e.g., copies of poster presentations), lack of individual data, inadequate methodological details, submission of very old data (>10 years) or data from completely different laboratories, use of inappropriate positive control chemicals or doses that were without effect, lack of statistical analysis, use of only one sex, and use of incompatibly aged animals. Analyses revealed that there were only 3 out of 16 laboratories that had submitted positive control data adequate for proficiency purposes for all of the major endpoints in the DNT study. Adequate positive control data are very useful in a weight-of-evidence approach to help determine the biological significance of results, and also to increase the confidence in negative results from DNT studies. PMID- 15113597 TI - Developmental alcohol exposure disrupts circadian regulation of BDNF in the rat suprachiasmatic nucleus. AB - In rats, damage to neuronal populations in some brain regions occurs in response to neonatal alcohol exposure coinciding with the period of rapid brain growth. These alcohol-induced defects in brain development may persist into adulthood and thus have long-term implications for the functional characteristics of damaged neuronal populations. The present study examined the effects of neonatal alcohol exposure on endogenous rhythmicity of the circadian clock located in the rat suprachiasmatic nucleus (SCN). Specifically, experiments were conducted to determine whether neonatal alcohol exposure alters the circadian rhythm of brain derived neurotrophic factor (BDNF) content in the rat SCN because this neurotrophin is an important rhythmic output from the SCN clock. Male rat pups were exposed to alcohol (4.5 g/kg/day) or isocaloric milk formula on postnatal days 4-9 using artificial rearing methods. At 5-6 months of age, SCN and hippocampal tissue was harvested and subsequently examined for content of BDNF protein. Time-dependent fluctuations in BDNF protein levels were assessed by enzyme-linked immunosorbent assay (ELISA). In alcohol-treated rats, SCN levels of BDNF were significantly decreased and were characterized by a loss of circadian rhythmicity relative to those observed in control animals. In comparison, hippocampal levels of BDNF displayed no evidence of circadian regulation in all three treatment groups, but were slightly lower in alcohol-treated animals than in control groups. Importantly, these observations suggest that alcohol exposure during the period of rapid brain development may cause permanent changes in the SCN circadian clock. PMID- 15113598 TI - Neurodevelopmental effects of postnatal lead exposure at very low levels. AB - This study is among the first to examine specific neurobehavioral deficits in children exposed at very low lead levels. A systematic analysis for the presence of a threshold of lead exposure was conducted. The sample consisted of 246 African American, inner-city children from whom blood lead concentrations were assessed at 7.5 years of age. The results consistently show neurobehavioral deficits in relation to low levels of lead in the areas of intelligence, reaction time, visual-motor integration, fine motor skills, attention, including executive function, off-task behaviors, and teacher-reported withdrawn behaviors. Effects were identified in the specific domains of attention, executive function, visual motor integration, social behavior, and motor skills, which have been previously suggested as part of lead's "behavioral signature". Visual inspection of nonparametric regression plots suggested a gradual linear dose-response relation for most endpoints. No threshold discontinuity was evident. Regression analyses in which lead exposure was dichotomized at 10 microg/dl were no more likely to be significant than analyses dichotomizing exposure at 5 microg/dl. Given that associations were found between lead levels as low as 3 microg/dl for multiple outcomes, these data provide additional evidence that there is no apparent lower bound threshold for postnatal lead exposure. PMID- 15113599 TI - Developmental effects of exposure to environmental tobacco smoke and material hardship among inner-city children. AB - Because of the growing concern that exposures to airborne pollutants have adverse effects on fetal growth and early childhood neurodevelopment, and the knowledge that such exposures are more prevalent in disadvantaged populations, we assessed the joint impact of prenatal exposure to environmental tobacco smoke (ETS) and material hardship on the 2-year cognitive development of inner-city children, adjusted for other sociodemographic risks and chemical exposures. The purpose was to evaluate the neurotoxicant effects of ETS among children experiencing different degrees of socioeconomic disadvantage, within a minority population. The sample did not include children exposed to active maternal smoking in the prenatal period. Results showed significant adverse effects of prenatal residential ETS exposure and the level of material hardship on 2-year cognitive development, as well as a significant interaction between material hardship and ETS, such that children with both ETS exposure and material hardship exhibited the greatest cognitive deficit. In addition, children with prenatal ETS exposure were twice as likely to be classified as significantly delayed, as compared with nonexposed children. Postnatal ETS exposure in the first 2 years of life did not contribute independently to the risk of developmental delay, over and above the risk posed by prenatal ETS exposure. The study concluded that prenatal exposure to ETS in the home has a negative impact on 2-year cognitive development, and this effect is exacerbated under conditions of material hardship in this urban minority sample. PMID- 15113600 TI - Monitoring protein phosphatase 1 isoform levels as a marker for cellular stress. AB - Reversible protein phosphorylation is a central mechanism regulating many biological functions, and abnormal protein phosphorylation can have a devastating impact on cellular control mechanisms, including a contributing role in neurodegenerative processes. Hence, many promising novel drug development strategies involve targeting protein phosphorylation systems. In this study, we demonstrate that various cellular stresses relevant to neurodegeneration can specifically affect the protein expression levels of protein phosphatase 1 (PP1). PP1 levels were altered upon exposure of PC12 and COS-1 cells to aluminium, Abeta peptides, sodium azide, and even heat shock. Particularly interesting, given PP1's involvement in aging and neurodegeneration, was the consistent decrease in PP1gamma(1) levels in response to stress agents. In fact, alterations in the expression levels of PP1 appear to correspond to an early response of stress induction, that is, before alterations in heat shock proteins can be detected. Our data suggest that monitoring PP1 isoform expression could constitute a useful diagnostic tool for cellular stress, possibly even neurodegeneration. Additionally, our results strengthen the rationale for signal transduction therapeutics and indicate that altering the specific activity of PP1 either directly or by targeting its regulatory proteins may be a useful therapeutic development strategy for the future. PMID- 15113601 TI - Assessment of PC12 cell differentiation and neurite growth: a comparison of morphological and neurochemical measures. AB - In vitro techniques are used increasingly to screen for and characterize neurotoxicants. In many cases, chemical-induced injury to developing neurons has been examined in vitro by assessing morphological changes in differentiation and neurite growth. This research evaluated the use of proteins associated with axonal growth and synaptogenesis as surrogates for morphological measurement of neuronal differentiation. PC12 cells, which differentiate upon nerve growth factor (NGF) stimulation, were used as the in vitro model. NGF-induced (50 ng/ml) differentiation (cells with at least one neurite with a length equal to the cell body diameter) and neurite growth (length of longest neurite) were determined using light microscopy and computer-based quantitative image analysis. PC12 cell differentiation and neurite growth reached a plateau after 6 days in culture. Expression of the axonal growth associated protein 43 (GAP-43) and the synaptic protein synapsin I were assessed simultaneously by Western blot during cell differentiation. Expression of GAP-43 was low on Culture Day 0 and increased progressively to maximum levels on Culture Day 5. Likewise, synapsin I expression increased slowly on Days 0-4, and then rapidly on Days 5-7 of culture. Pharmacologic inhibitors of NGF-induced signaling were used to test the sensitivity of the proteins to chemical disruption of differentiation. The MAP kinase inhibitor, U0126 (5-30 microM) and the PKC inhibitor, bisindolylmaleimide I (Bis I; 1.25-5 microM) inhibited differentiation and neurite outgrowth in a concentration-dependent manner. U0126 and Bis I significantly decreased GAP-43, but not synapsin I expression. Interestingly, the PI-PLC inhibitor edelfosine (ET 18; 5-30 microM) stimulated differentiation at early times of exposure followed by a significant decrease in neurite length at later time points. However, ET-18 did not alter the expression of GAP-43 or synapsin I. These data suggest that GAP 43 may be a useful indicator of the status of PC12 cell differentiation. PMID- 15113602 TI - Differential profiles of cholinesterase inhibition and neurobehavioral effects in rats exposed to fenamiphos or profenofos. AB - The relationship between cholinesterase (ChE) inhibition and neurobehavioral changes was examined using two ChE-inhibiting organophosphorus (OP) pesticides, fenamiphos and profenofos. Both pesticides produce considerable blood ChE inhibition, but relatively little brain inhibition up to almost lethal doses. Interestingly, pronounced neurobehavioral signs were produced by fenamiphos but not profenofos. After a single oral dose, both pesticides greatly inhibited blood ChE (87-98% inhibition), yet whole brain ChE was only inhibited by 9-14% at the highest doses. Fenamiphos produced dose-dependent effects on many behavioral measures. Despite the similar ChE inhibition profile, profenofos produced no observable changes in behavior. Treatment with anticholinergic drugs was used to evaluate the contribution of peripheral versus central ChE inhibition. Scopolamine (SCO) and methylscopolamine (MSC) were used as central/peripheral and peripheral-only cholinergic receptor blockers, respectively, in combination with fenamiphos. Neither drug altered the effects of fenamiphos on ChE inhibition. Some behavioral effects of fenamiphos were blocked or attenuated only by SCO, whereas other effects were blocked by both drugs. These data indicate that some of the pronounced neurobehavioral changes observed following fenamiphos dosing may be centrally mediated (blocked by SCO only), despite the small amount of inhibition of brain ChE. Other behavioral changes may be mediated more peripherally (blocked by both MSC and SCO). To test the hypothesis that regionally specific ChE inhibition may be responsible for these effects, the same dose of fenamiphos used in the previous studies was given and one half of the brain was dissected into regions. There was significant ChE inhibition in the pons and medulla, cerebellum, striatum, hippocampus, and half-brain but not in the rest-of-brain and frontal cortex; however, the magnitude of inhibition was relatively small across the regions measured. Thus, the centrally mediated neurobehavioral effects of fenamiphos could not be explained based on differential regional brain ChE inhibition alone. Despite the low level of brain ChE inhibition, some behavioral effects of fenamiphos were centrally mediated, and there was little regional specificity of ChE inhibition that could account for the behavioral changes observed. PMID- 15113603 TI - Prenatal ethanol enhances rotational behavior to apomorphine in the 24-month-old rat offspring with small striatal lesion. AB - Pregnant Wistar rats received a hyperproteic liquid diet containing 37.5% ethanol derived calories during gestation. Isocaloric amount of liquid diet, with maltose dextrin substituted for ethanol, was given to control pair-fed dams. Offsprings were allowed to survive until 24 months of age. A set of aged female offsprings of both control diet and ethanol diet groups was registered for spontaneous motor activity, by means of an infrared motion sensor activity monitor, or for apomorphine-induced rotational behavior, while another lot of male offsprings was submitted to an unilateral striatal small mechanical lesion by a needle, 6 days before rotational recordings. Prenatal ethanol did not alter spontaneous motor parameters like resting time as well as the events of small and large movements in the aged offsprings. Bilateral circling behavior was already increased 5 min after apomorphine in the unlesioned offsprings of both the control and ethanol diet groups. However, it lasted more elevated for 45- to 75-min time intervals in the gestational ethanol-exposed offsprings, while decreasing faster in the control offsprings. Apomorphine triggered a strong and sustained elevation of contraversive turns in the striatal-lesioned 24-month-old offsprings of the ethanol group, but only a small and transient elevation was seen in the offsprings of the control diet group. Astroglial and microglial reactions were seen surrounding the striatal needle track lesion. Microdensitometric image analysis demonstrated no differences in the levels of tyrosine hydroxylase immunoreactivity in the striatum of 24-month-old unlesioned and lesioned offsprings of control and alcohol diet groups. The results suggest that ethanol exposure during gestation may alter the sensitivity of dopamine receptor in aged offsprings, which is augmented by even a small striatal lesion. PMID- 15113604 TI - Prenatal cocaine exposure influences the growth and life span of laboratory rats. AB - Laboratory rats prenatally exposed to alcohol, nicotine, amphetamine, undernutrition or hypoxia can exhibit shortened life span and other signs of enhanced age-related degeneration. We evaluated the possibility of similar effects following prenatal cocaine exposure. Pregnant rats received 20 or 40 mg/kg cocaine HCl subcutaneously (C20, C40), twice daily, from gestation days (GD) 7-20. Untreated control (UTC) and pair-fed control (PFC) groups were also used. The pregnant C40, C20, and PFC dams ate less food and gained less weight than the UTC dams did. The pregnant C40 and C20 dams drank more water than the UTC dams did, and the pregnant PFC dams drank less than the UTC dams did. The C40 and PFC offspring had delayed earflap openings. The C40 male and female offspring had lower birth weights than their cohorts in the other three groups. The C40 female and male offspring remained significantly underweight until postnatal day (PND) 28 and PND56, respectively. During young adulthood, the males and females in the C20, C40, and PFC groups had normal body weights. During old adulthood, however, the C20 and C40 males and the C20, C40, and PFC females developed reduced body weights as compared with their UTC cohorts. The C20 and C40 male offspring and the C20, C40, and PFC female offspring also had life spans that were 7-12% shorter than that of their UTC cohorts. Thus, groups that showed reduced body weights in old age also showed shorter life spans. These results provided converging evidence that prenatal cocaine exposure caused enhanced age related degeneration. Observations on cardiac and other organ pathology were also made. Health implications for children born to cocaine-abusing women are discussed. PMID- 15113605 TI - Perinatal cocaine exposure reduces myocardial norepinephrine transporter function in the neonatal rat. AB - Norepinephrine transporter (NET) mediates the active removal of norepinephrine (NE) released from sympathetic nerve terminals via reuptake, and NET function and expression can be regulated by cocaine. NET expression and its regulation by cocaine in the developing sympathetic nervous system during early postnatal period, however, have not been examined. We quantified immunodetectable NET protein expression in the neonatal rat heart to examine the developmental pattern of myocardial NET during the first 2 weeks after birth. To assess sympathetic innervations, we simultaneously quantified the expression of myocardial tyrosine hydroxylase (TH). Timed pregnant rats received daily intragastric treatment with saline (CTL) or cocaine at 60 mg/kg (Coc) from Gestational Day 2 until parturition. After birth, nursing mothers continued to receive the same treatment. The expression of myocardial TH and NET in neonatal rats were then studied at 1 day (Postnatal Day 1, PD1), 7 days (PD7) or 14 days (PD14) of age. We observed a similar age-dependent increase in the expression for myocardial NET and TH during the first 2 weeks of postnatal life, in both CTL and Coc animals. While myocardial TH was significantly up-regulated following perinatal cocaine exposure, no significant change in immunodetectable myocardial NET protein was evident. To further examine whether NET function might be affected by perinatal cocaine exposure, we performed NE uptake in myocardial membranes from PD14 CTL and Coc rats. We found that NE uptake was reduced at PD14 in the cocaine-treated group. Our results indicate that myocardial NET and TH are both developmentally regulated. Furthermore, our results indicate that perinatal exposure to cocaine did not change NET protein expression but impaired myocardial NET function in the neonatal rat. PMID- 15113606 TI - Modality-specific impairments in response habituation following postnatal binge ethanol. AB - The rate of habituation of the heart rate orienting response (OR) to both olfactory and auditory cues was examined in subjects neonatally exposed to ethanol. Subjects were administered 5.25 g/kg/day ethanol in a binge-like manner on either postnatal days (PDs) 4-9 or 10-15. Controls were given sham intubations. Ethanol exposure had no effect on the form or magnitude of the heart rate OR; all subjects exhibited a monophasic bradycardia to the olfactory and auditory cues. However, ethanol treatment severely affected the rate of response habituation to an olfactory cue, regardless of the timing of that exposure. Ethanol treatment on either PDs 4-9 or 10-15 resulted in a failure of animals to show complete heart rate response habituation. For the auditory cue, rate of response habituation was unaffected by ethanol treatment. These results indicate that response habituation deficits in a postnatal model of human fetal-alcohol exposure are modality specific. The response habituation paradigm may be useful for studying mechanisms of short-term recognition memory deficits following early alcohol exposure, although modality of the test stimulus needs to be taken into account. The results suggest that central nervous system structures affected by ethanol may be specific to those involved in olfactory processing, such as the olfactory bulb or hippocampus. PMID- 15113607 TI - Developmental aspects of timing behavior in children. AB - This research examined the association of age, sex, and intelligence on the performance of a time production (temporal response differentiation, TRD) task. Variations of this task have been used extensively with both animals and humans to study factors that affect aspects of timing ability. The participants in this study (720 children, ages 5 to 13 years) were required to hold down a response lever for at least 10 s, but no more than 14 s, to receive a nickel. Older children made more correct lever holds and exhibited less variability in the duration of their lever holds than did the younger children. Boys and girls performed similarly on this task, whereas children with higher IQs made more correct lever holds. Young children with below average IQs exhibited increased variability in lever hold duration compared with young children with average and above average IQs. The results of this study illustrate that both age and intelligence influence timing ability. The use of this timing task in children, which also has been widely used in animal models, provides unique opportunities for interspecies comparisons. PMID- 15113608 TI - Deferoxamine antioxidant activity on cerebellar granule cells gamma-irradiated in vitro. AB - Oxidative stress has been implicated in the pathogenesis of many neurodegenerative and neurological disorders, with reactive oxygen species (ROS) as part of the intracellular effectors of damage formed in the presence of an excess of iron. Ionizing radiation induces tissue damage on developing CNS through different simultaneous mechanisms, including ROS-induced oxidative damage; therefore, exogenously added iron chelators might contribute to protect cells from free-radical injury. Cerebellar granule cells grown in vitro were exposed to 0.3 Gy of gamma radiation, and 30-60 min before irradiation, deferoxamine (Dfx), an iron chelator, was added at different nontoxic concentrations. When cell viability and ROS levels were evaluated in Dfx-treated cultures, a partial prevention of radiation-induced cell death and ROS increase were found, being this prevention concentration independent. These data support the involvement of an iron-driven hydroxyl radical formation pathway in the acute toxic mechanism of radiation in cultures of cerebellar granule cells, being ROS induced oxidative damage one of the mechanisms through which radiation might induce cell death. Therefore, blocking ROS production through the use of a chelating agent, such as Dfx, would be a useful therapeutic tool in different experimental models. PMID- 15113609 TI - Effect of lactide/glycolide monomers on release behaviors of gentamicin sulfate loaded PLGA discs. AB - In order to develop the desirable drug release patterns such as no lag time and exact zero-order release rate, gentamicin sulfate (GS)-loaded poly(D,L-lactide-co glycolide) (PLGA) discs containing lactide monomer (LM) or glycolide monomer (GM) were prepared. LM or GM was applied for the controlling drug release pattern due to its non-toxic and biodegradable nature. Water absorption, mass loss, pH change, and morphology of discs were examined to study the effect of LM or GM addition. GS release showed near zero-order profile in the GS-loaded polymeric discs prepared in the presence of LM or GM (10%). The channel of GS-loaded PLGA containing LM or GM was formed by the dissolution of LM or GM. Water uptake of disc increased till 21 days from the beginning of the test. The pH variations of media declined in the same manner with the result of mass loss. The antibiosis of GS was also confirmed by bacterial inhibition zone test using the prepared polymeric discs. From these results, we expected that the polymeric discs containing LM or GM would be a good dosage form as a topically implantable device which can get rid of lag period from PLGA matrix. PMID- 15113610 TI - Transbuccal permeation, anti-inflammatory activity and clinical efficacy of piroxicam formulated in different gels. AB - In attempts to avoid the systemic side effects of piroxicam (PC) (e.g. gastrotoxicity), several buccal gel formulations containing PC were prepared and their effects on the characteristics of the drug permeation through rabbit buccal mucosa in-vitro were evaluated using a Franz-type diffusion cell. The general rank order of the total flux of 0.5% PC from gels was found to be: hydroxypropylmethylcellulose (HPMC, 2.5%) > hydroxypropylcellulose (HPC, 2.5%) >or= sodium alginate (Na alg., 7%) > methylcellulose (MC, 3%) > hydroxyethylcellulose (HEC, 1.5%) > carbopol 934 (Carb. 934, 1%) >or= sodium carboxymethylcellulose (NaCMC, 2%) > pluronic F-127 (PF-127, 20%) > polyvinyl alcohol (PVA, 10%). The effect of various penetration enhancers 1% sodium lauryl sulphate (NaLS), 3% sodium deoxycholate (NaDC), 3% sodium tauroglycocholate (NaTGC) on the rate of permeation across the excised buccal mucosa (of 0.5% PC in gels prepared using 3% MC, 2.5% HPMC or 7% Na alg. base) and histology of the buccal epithelium was also investigated. Pharmacodynamic evaluation of the anti inflammatory activity of PC in these gel formulations (containing 3% NaDC as an enhancer) was carried out using the kaolin-induced rat paw oedema method. The results obtained indicated that PC administered in 7% Na alg. or 2.5% HPMC gel bases was significantly more effective than the 3% MC gel and oral drug solution in suppressing oedema formation in rats. Comparative clinical studies were conducted in patients with post-operative dental pain and oedema following maxillofacial operations. The results revealed that 7% Na alg. and 2.5% HPMC gel formulations applied to the buccal mucosa were slightly better than or equally effective to the orally administered commercial product (Feldene Flash) tablet) in reducing pain level, swelling and tenderness within a period of 4 days. These findings suggest that PC (0.5%) administered in the buccal gel may present a potential therapeutical use as a strong anti-inflammatory and analgesic agent. PMID- 15113611 TI - A rabbit ear flap perfusion experiment to evaluate the percutaneous absorption of drugs. AB - A rabbit ear flap single-pass perfusion system was examined as an experimental method for studying the relationship between the physiological conditions of tissues and drug disposition after topical applications. Tyrode solutions containing bovine serum albumin (BSA) and sucrose or flurbiprofen (FP), used as a model drug, were perfused through the vessel in the ear flap to evaluate the physiological conditions prior to the application of FP to the skin surface. The extracellular volume and distribution properties of FP in the perfused ear were similar to those in an in vivo experimental system. In addition, the perfused ear flap exhibited a pharmacological response to bradykinin (BK). The amount of FP in the outflow Tyrode solution containing BSA after application to the skin surface of the perfused ear decreased with the addition of BK, while that in the tissues under the application site increased. FP binds to BSA, which leaked from the intravascular space, and could be retained in the tissues under the application site. The protein binding also affected the redistribution of FP to other tissues in the ear flap after application to the skin. The rabbit ear perfusion system is a useful method for studying the percutaneous absorption of drugs especially variations in the disposition of drugs in oedematous tissues. PMID- 15113612 TI - Comparative bioavailability of rifampicin, isoniazid and pyrazinamide from a four drug fixed dose combination with separate formulations at the same dose levels. AB - Fixed dose combination (FDC) formulations became popular in the treatment of tuberculosis (TB) because of the better patient compliance, reduced risk of monotherapy and emergence of drug resistance in contrast to treatment with separate formulations of two to four first-line drugs. However, its successful implementation in national programs is limited by probable bioinequivalency of rifampicin if present in FDC form. In this regard, World Health Organization (WHO) and International Union Against Tuberculosis and Lung Disease (IUATLD) recommend FDCs only of proven bioavailability. Hence, bioequivalence study of four drug FDC tablet was conducted using 22 healthy male volunteers according to WHO recommended protocol to determine bioavailability of rifampicin, isoniazid and pyrazinamide compared to standard separate combination at the same dose level. The study was designed as two period, two treatment crossover experiment with a washout period of 1 week. Bioequivalence of rifampicin was estimated by plasma and urinary method for both rifampicin and its active metabolite, des acetyl rifampicin whereas isoniazid and pyrazinamide were estimated from plasma. Mean concentration time profiles and all the pharmacokinetic parameters of rifampicin, isoniazid and pyrazinamide from FDC tablet were comparable to individual formulations and passed the bioequivalence test with power of the test above 95%. Further, bioequivalence of both rifampicin and isoniazid shows that in vitro interaction of rifampicin and isoniazid is clinically insignificant. Thus, it was concluded that FDC formulation is bioequivalent for rifampicin, isoniazid and pyrazinamide and ensures the successful treatment of TB without compromising therapeutic efficacy of any of these components of anti-TB therapy. PMID- 15113613 TI - In situ evaluation of drug-loaded microspheres on a mucosal surface under dynamic test conditions. AB - The ability of polymeric microspheres fabricated from Carbopol, polycarbophil, chitosan or Gantrez to retain a model hydrophilic drug (sodium fluorescein) was evaluated in situ, using a dynamic test system and image analysis. This technique used oesophageal tissues and simulated the physiological conditions within the oral cavity in terms of temperature, humidity and saliva flow. The point of sample application was observed over a 2h period by means of a digital camera. No significant differences in fluorescein colour intensity was obtained for the Gantrez and chitosan particles over 100min, indicate that these two polymers provide the possibility of prolonged action. Carbopol and polycarbophil particles became rapidly swollen and released the sodium fluorescein completely within 20min. It was concluded that the test system allowed the evaluation of the in situ behaviour of test particles, in terms of their ability to retain a water soluble, coloured marker in 'dynamic' test conditions, and that chitosan and Gantrez were promising candidates for the production of mucoadhesive, sustained release microspheres for water-soluble materials. PMID- 15113614 TI - Drug release from carbomer:carbomer sodium salt matrices with potential use as mucoadhesive drug delivery system. AB - In vitro mucoadhesion, water uptake, and drug release of nystatin (N) from matrices of carbomer (C) and lyophilized carbomer sodium salt (CNaL) mixtures were evaluated. Matrices with different ratios C:CNaL were prepared by direct compression. Commercial C as well as lyophilized powder (CL) were used. In vitro mucoadhesion increased as the proportion of C in the matrix was raised. The same effect was observed when C was replaced by CL. Matrices in which C was replaced by CL showed an increase of both water uptake and release rates. Besides, the release of N from matrices CL:CNaL exhibited a kinetics with Super Case II (n>1) mechanism. However, for C:CNaL matrices, drug release was slower and exhibited a biphasic profile with a first stage characterized by either an anomalous (n<1, for C>or=50%) or a Case II (n approximately 1.0, C<50%) mechanisms. After that period, the mechanism changed to Super Case II transport (n>1). PMID- 15113615 TI - Cell type-specific gene expression, mediated by TFL-3, a cationic liposomal vector, is controlled by a post-transcription process of delivered plasmid DNA. AB - The issue of whether the TFL-3, a recently developed cationic liposome, achieves efficient gene expression in different mammalian cell lines (NIH/3T3, LLC, A431 and HeLa cells) was examined. The issue of whether gene expression is related to the amount of plasmid DNA (pDNA) delivered in cells or nuclei following transfection was also examined. The cells were transfected for 1h with pDNA/TFL-3 lipoplexes, and the transfection efficiency was determined by means of a luciferase activity assay. The amount of intracellular and intranuclear pDNA following the transfection was also quantitatively determined. Successful transgene expressions in all cell lines we tested were observed under our experimental conditions, suggesting that the TFL-3 represents a suitable nonviral vector system for the successful gene expression in mammalian cells in vitro. The degree and rate of gene expression were dependent on the type of cells used as well as the incubation time after transfection, but these parameters were independent of the amount of gene delivered to cells and nuclei. These results suggest that TFL-3 mediated gene expression is largely controlled by the process of post-transcription of the delivered pDNA, and not by the process of cellular entry of pDNA and cytoplasmic trafficking of pDNA into nuclei, which is dependent on the cell type. Therefore, the results obtained here clearly suggest that the cell type-specific improvement in transcription efficiency of pDNA and translation of the derived mRNA, together with an improved delivery system to enhance the nuclear delivery of pDNA, is necessary to achieve efficient transgene expression in mammalian cells. PMID- 15113616 TI - Pulmonary delivery of TH9507, a growth hormone releasing factor analogue, in the dog. AB - A modified growth hormone releasing factor (GRF; TH9507), a 44 amino acid peptide analogue of natural human growth hormone releasing factor, is being developed for the treatment of age-associated conditions resulting from diminished growth hormone (GH) secretion. The inhalation route of administration is being considered as an alternative to subcutaneous injection. A study was undertaken in dogs to investigate the absorption of TH9507 following pulmonary delivery. Male beagle dogs were administered TH9507 by intratracheal dry powder insufflation and subcutaneous injection at doses of approximately 375 and 38microg/kg, respectively. In a separate study, male and female dogs received 100microg/kg intravenously. Blood samples were collected at selected sampling times after dosing and plasma levels of TH9507 were measured by radioimmunoassay. The bioavailability by the inhaled route was 41% relative to subcutaneous dosing, with an absolute bioavailability estimated at 13%. No significant difference was observed for the terminal half-life of TH9507 after intratracheal (39min) and subcutaneous (26min) administrations. The mean residence time (MRT) was greater following intratracheal administration (74min versus 52min; P<0.01). These data indicate that the delivery of the TH9507 by the inhalation route may provide a suitable alternative to subcutaneous injection. PMID- 15113617 TI - A novel in situ gel for sustained drug delivery and targeting. AB - The objective of this study was to develop a novel chitosan-glyceryl monooleate (GMO) in situ gel system for sustained drug delivery and targeting. The delivery system consisted of 3% (w/v) chitosan and 3% (w/v) GMO in 0.33M citric acid. In situ gel was formed at a biological pH. In vitro release studies were conducted in Sorensen's phosphate buffer (pH 7.4) and drugs were analyzed either by HPLC or spectrophotometry. Characterization of the gel included the effect of cross linker, determination of diffusion coefficient and water uptake by thermogravimetric analysis (TGA). Mucoadhesive property of the gel was evaluated in vitro using an EZ-Tester. Incorporation of a cross-linker (glutaraldehyde) retarded the rate and extent of drug release. The in vitro release can further be sustained by replacing the free drug with drug-encapsulated microspheres. Drug release from the gel followed a matrix diffusion controlled mechanism. Inclusion of GMO enhanced the mucoadhesive property of chitosan by three- to sevenfold. This novel in situ gel system can be useful in the sustained delivery of drugs via oral as well as parenteral routes. PMID- 15113618 TI - Formulation and characterization of Paclitaxel, 5-FU and Paclitaxel + 5-FU microspheres. AB - The purpose of this study was to compare the combination (Paclitaxel + 5-FU microspheres) with a single drug chemotherapy (Paclitaxel and 5-FU microspheres) against metastatic breast cancer cell line (MDA-MB 435 S). The physicochemical characteristics of the microspheres (i.e. encapsulation efficiency, particle size distribution, in vitro release, thermal characteristics) were studied. The results demonstrated that the encapsulation efficiency of Paclitaxel was high (90%) when the drug was encapsulated in poly(lactic-co-glycolic acid) (PLGA) microparticles with or without 5-fluorouracil (5-FU). However, the encapsulation efficiency of 5-FU was low (19%) and increased to 30% when the drug was encapsulated with Paclitaxel. The mean particle size of microspheres was 2.5microm and were spherical in shape. The in vitro release of both 5-FU and Paclitaxel from the microspheres was relatively fast initially followed by a slower and more controlled release. The cytotoxic activity of Paclitaxel microspheres was far greater compared to either the microspheres containing 5-FU + Paclitaxel or 5-FU alone. Overall results demonstrated that incorporation of Paclitaxel or 5-FU in microspheres enhances the cytotoxicity in more controlled manner compared to that of free drugs and also that careful consideration should be made when combining drugs acting in different phases of cell cycle. PMID- 15113619 TI - Modeling of drug release from partially coated matrices made of a high viscosity HPMC. AB - A mathematical model able to describe the release kinetics of two model drugs (Diprophylline and Theophylline) from partially coated hydroxypropylmethylcellulose (HPMC, Methocel) K4M) matrices is presented. As solvent interaction with the system and drug release can only take place in one direction, the physical frame to be modeled turns out simpler. The model was developed starting from the established equation describing drug dissolution and taking into account the resistance to drug release given by the presence of a growing gel barrier around a matrix system. The model fits the release data obtained from both series of hydrophilic matrices containing increasing amounts (from 0.2 to 0.8 mass ratio) of the two xanthine derivatives. Differences were found in drug release rate according to the different solubility of the actives. Interestingly, however, there is no further reduction in the outer gel layer permeability when the polymer mass fraction exceeds a certain value, with both Theophylline and Diprophylline systems. Results confirm the importance of the fraction of the glassy/rubbery interface held by the active substance in defining the release rate from hydrophilic systems. PMID- 15113620 TI - Structural characterisation of water-Tween 40/Imwitor 308-isopropyl myristate microemulsions using different experimental methods. AB - Pharmaceutically usable microemulsion systems were prepared from water and isopropyl myristate with a constant amount of Tween 40 and Imwitor 308 at a mass ratio of 1. Their type and structure were examined by measuring density and surface tension, and by viscometry, electric conductivity, differential scanning calorimetry (DSC) and small-angle X-ray scattering (SAXS), and the degree of agreement between the techniques was assessed. A model based on monodisperse hard spheres adequately fits the SAXS data in W/O microemulsions predicting, depending on composition, elongated or spherical droplets. It also suggests the involvement of strong attractive interactions in O/W systems. Results of conductivity, viscosity, density and surface tension measurements confirm the prediction of a percolation transition to a bicontinuous structure. DSC detects the degree of water interaction with surfactants thus identifying the type of microemulsion. The conclusions from all the techniques agree well and indicate that such studies could also be carried out on more complex systems. In future, the ability to determine type and structure of such microemulsion systems could enable partitioning and release rates of drugs from microemulsions to be predicted. PMID- 15113621 TI - Comparison of the effects of two drying methods on polymorphism of theophylline. AB - Processing-induced transformations in drug formulation may induce adverse biopharmaceutical changes in the finished product. During the drying phase of wet granulation, theophylline monohydrate transforms either the stable (form I), or a polymorphic, metastable (form I(*)) form of anhydrous theophylline. We investigated the effect of two drying methods (multichamber microscale fluid bed dryer MMFD) or variable temperature X-ray powder diffractometer (VT-XRPD) on the relative amounts of the different theophylline forms remaining in the dried granules. Granules were analyzed using XRPD and near-infrared spectroscopy. Form I(*) was the predominant form of theophylline after drying at 40-50 degrees C with both drying techniques. Although drying at temperatures over 50 degrees C produced mostly form I, more than 20% of form I(*) remained even at 90 degrees C when drying in MMFD. In these conditions, humidity had little influence on the amount of form I(*) in the granules. In contrast, drying in a VT-XRPD at 60 degrees C produced form I already during the first 15min. Using additional drying methods, including MMFD, during the preformulation stage can be more informative about the possible polymorphic transformations and their underlying mechanisms, such as triboelectrification or recrystallization, in drug ingredients during the manufacturing process. PMID- 15113622 TI - Interactions of surfactants (edge activators) and skin penetration enhancers with liposomes. AB - Incorporating edge activators (surfactants) into liposomes was shown previously to improve estradiol vesicular skin delivery; this phenomenon was concentration dependent with low or high concentrations being less effective. Replacing surfactants with limonene produced similar behaviour, but oleic acid effects were linear with concentration up to 16% (w/w), beyond which it was incompatible with the phospholipid. This present study thus employed high sensitivity differential scanning calorimetry to probe interactions of additives with dipalmitoylphosphatidylcholine (DPPC) membranes to explain such results. Cholesterol was included as an example of a membrane stabiliser that removed the DPPC pre-transition and produced vesicles with a higher transition temperature (T(m)). Surfactants also removed the lipid pre-transition but reduced T(m) and co operativity of the main peak. At higher concentrations, surfactants also formed new species, possibly mixed micelles with a lower T(m). The formation of mixed micelles may explain reduced skin delivery from liposomes containing high concentrations of surfactants. Limonene did not remove the pre-transition but reduced T(m) and co-operativity of the main peak, apparently forming new species at high concentrations, again correlating with vesicular delivery of estradiol. Oleic acid obliterated the pre-transition. The T(m) and the co-operativity of the main peak were reduced with oleic acid concentrations up to 33.2mol%, above which there was no further change. At higher concentrations, phase separation was evident, confirming previous skin transport findings. PMID- 15113623 TI - Fatty acids in Botryococcus braunii accelerate topical delivery of flurbiprofen into and across skin. AB - To improve the drug absorption into and across the skin, fatty acids extracted from Botryococcus braunii were evaluated using in vitro and in vivo techniques with Wistar rats as the animal model. Palmitic acid (C16:0), oleic acid (C18:1), linoleic acid (C18:2), and linolenic acid (C18:3) were the major components in the B. braunii extract. Topical delivery of flurbiprofen was significantly enhanced after pretreatment with 3% B. braunii extract for 30min in an in vitro Franz cell and in vivo pharmacokinetic studies. Pure unsaturated fatty acids were more-effective enhancers than the B. braunii extract. However, a greater irritant potential was also observed with those fatty acids than with the B. braunii extract according to the skin tolerance study as determined by transepidermal water loss (TEWL). Both human keratinocytes and skin fibroblasts showed a 1.5-2 fold increase in prostaglandin E(2) (PGE(2)) release as compared to the control. The findings in this study indicate that the fatty acids in B. braunii may be useful enhancers for flurbiprofen delivery via the skin. PMID- 15113624 TI - Phospholipid-based microemulsion formulation of all-trans-retinoic acid for parenteral administration. AB - All-trans-retinoic acid (ATRA) shows anti-cancer activities, especially in patients with acute promyelocytic leukemia. Due to the highly variable bioavailability of ATRA and induction of its own metabolism after oral treatment, development of alternative parenteral dosage form is required. The principal aim of this study was to develop a parenteral formulation of ATRA by overcoming its solubility limitation by utilizing phospholipid-based microemulsion system as a carrier. Microemulsion was prepared with pharmaceutically acceptable ingredients such as soybean oil and phospholipids. The mean particle diameter and polydispersity of ATRA microemulsion could be decreased to be applicable for parenteral administration by modulation of composition of microemulsion. The loading concentration of ATRA in microemulsion increased by increasing the oil contents and also by inclusion of distearoylphosphatidyl-ethanolamine-N poly(ethyleneglycol) 2000 (DSPE-PEG). Furthermore, loading of ATRA in microemulsion improved the chemical stability of ATRA. The pharmacokinetic profile of ATRA after intravenous injection of microemulsion formulation to rats was similar to that of sodium ATRA. The growth inhibitory effects of ATRA on human cancer HL-60 and MCF-7 cell lines were also similar between free ATRA and microemulsion formulation of ATRA, suggesting that its anti-cancer activity was not impaired by loading in microemulsion. Our study herein demonstrates that phospholipid-based microemulsion may provide an alternative parenteral formulation of ATRA. PMID- 15113625 TI - The production of homogeneous extrudates of microcrystalline cellulose pastes. AB - The homogeneity of water-based microcrystalline cellulose (MCC) paste extrudates was investigated during ram extrusion as a function of ram velocity. Variations in the water content of the extrudates were caused by liquid phase migration within the paste. The evolution in water content was measured by sectioning and drying the extrudate, and the subsequent homogeneity was quantified by the standard error in water content. The homogeneity of the extrudates was found to decrease as the ram velocity decreased. This result was also inferred from the rate of increase of the extrusion pressure. The extrudate homogeneity was significantly improved by compensating for water migration in the barrel during the compaction stage. This was achieved using a non-uniform initial paste billet, created by packing the barrel with layers of paste of different water contents. This technique also produced a smaller variation in extrusion pressure over the ram displacement range, and a reduction in water loss from the upstream paste compact into the extrudate and/or through the apparatus tooling. PMID- 15113626 TI - Sonographic morphology and hyaluronan content of umbilical cords of healthy and Down syndrome fetuses in early gestation. AB - OBJECTIVE: To explore the sonographic vascular architecture and the hyaluronan amount and distribution of umbilical cords of healthy and trisomy 21 fetuses in early gestation. MATERIAL AND METHODS: Umbilical cord sonographic morphology and morphometry of 112 consecutive normal fetuses and 11 trisomy 21 fetuses were assessed between 10 and 15 weeks of gestation. The umbilical coiling index was defined as the reciprocal of the length of one complete coil measured in a longitudinal section of the umbilical cord. The umbilical coiling angle was defined as the maximum angle between the long axis of the umbilical cord and that of the umbilical arteries. Three umbilical cord samples obtained from Down syndrome fetuses and one from a healthy fetus after voluntary termination of pregnancy at 13 weeks of gestation were used for biochemical analysis. Quantitative hyualuronan content and tissue distribution was studied using fluorophore-assisted carbohydrate electrophoresis (FACE) analysis and staining methods using biotin-labeled hyaluronan-binding protein (bHABP), respectively. RESULTS: A significant correlation was present between gestational age and both the umbilical coiling index (r=-0.56, p<0.001) and the umbilical coiling angle (r=-0.43, p<0.001). The proportion of uncoiled umbilical cords was significantly higher in Down syndrome fetuses than in healthy fetuses [8/112 (7.1%) vs. 4/11 (36.4%), p<0.05]. Biochemical analysis demonstrated a higher amount and a different distribution of hyaluronan in trisomy 21 umbilical cords compared to healthy fetuses. CONCLUSION: The umbilical cord of Down syndrome fetuses in early gestation shows peculiar sonographic vascular features and quantitative alterations of the Wharton's jelly hyaluronan. PMID- 15113627 TI - Impaired left ventricular diastolic function in newborn infants of mothers with pregestational or gestational diabetes with good glycemic control. AB - THE AIM OF THE STUDY: We assessed by echocardiography the left ventricular systolic and diastolic function in newborn infants of mothers with well controlled pregestational type 1 or gestational diabetes (IDM) in comparison to normal term neonates. SUBJECTS AND METHODS: Two-dimensional/M-mode and Doppler transmitral flow velocity measurements were performed in 18 IDM and 26 control infants of non-diabetic mothers (gestational ages 36-40 and 36-41 weeks, respectively) between days 2 and 5 after birth. In the IDM, there were nine mothers with pregestational (White class C or D) and nine mothers with gestational diabetes (White class A or A/B). Peak early and atrial filling velocity, early deceleration time, early acceleration time, early, atrial and total time velocity integrals were used to examine the left ventricular diastolic performance. We also calculated the early/atrial velocity ratio, early/atrial integral ratio and early/total integral ratio. The fractional shortening, fractional shortening area, midwall fractional shortening (mFS), left ventricular mass and indexed left ventricular mass for body surface area (BSA) and birth weight were used in assessment of left ventricular systolic performance. RESULTS: The early deceleration time was longer, resulting in higher early integral and early filling fraction (EFF) in the IDM than in the control infants (p<0.01). In the IDM, the fractional shortening was somewhat greater and the left ventricular mass/body surface area ratio was higher than in the control group (p<0.05), although the measures of systolic performance were within the normal range. There were no significant differences in the systolic or diastolic function parameters between the gestational and pregestational groups. CONCLUSION: In the infants of mothers with well-controlled pregestational or gestational diabetes, we found prolonged deceleration time of early left ventricular diastolic filling, probably reflecting an impaired left ventricular relaxation rather than compliance. The mechanism for the findings may be maternal hyperglycemia during the third trimester and subsequent fetal hyperinsulinaemia leading to neonatal cardiac hypertrophy. PMID- 15113628 TI - 2nd to 4th digit ratios, fetal testosterone and estradiol. AB - BACKGROUND: The ratio of 2nd to 4th digit length (2D:4D) is sexually dimorphic (mean 2D:4D is lower in males than females) and is thought to be fixed early in development. 2D:4D has been reported to be related to fetal growth, hand preference, autism, Asperger's syndrome, sperm counts, family size, age at myocardial infarction in men and breast cancer in women. There is indirect evidence that 2D:4D is established in utero and is negatively related to prenatal testosterone and positively with prenatal estradiol. However, there are no studies which show direct relationships between fetal testosterone (FT), fetal estradiol (FE) and 2D:4D. AIMS: To investigate the relationships between 2D:4D ratios and FT and FE from amniotic fluid. STUDY DESIGN: Cohort study. SUBJECTS: 33 children. OUTCOME MEASURES: Radioimmunoassays of FT and FE obtained from routine amniocentesis; 2D:4D ratios calculated from 2nd and 4th digit length of the right and left hands at age 2 years. RESULTS: A significant negative association between right 2D:4D ratio and FT/FE ratio, which was independent of sex. CONCLUSIONS: These preliminary findings lend support to an association between low 2D:4D and high levels of FT relative to FE, and high 2D:4D with low FT relative to FE. PMID- 15113629 TI - Association of cord blood cytokines with prematurity and cerebral palsy. AB - BACKGROUND: Many studies have shown that certain cytokines in amniotic fluids are correlated to premature labor and neonatal brain insults. AIMS: We investigated whether different fetal phagocyte and vascular mediators including IL-8, myeloperoxidase (MPO), PGE(2) and soluble vascular cell adhesion molecule-1 (sVCAM-1) levels were correlated to prematurity and cerebral palsy (CP) of premature infants. SUBJECTS: Umbilical cord blood samples from 96 preterm babies from 2250 cord blood collections were studied. METHOD: The enzyme-linked immunoassay (ELISA) technique was used to determine concentrations of cord blood IL-8, MPO, PGE(2) and sVCAM-1. RESULTS: Preterm babies with gestational age (GA) < or =32 weeks had significant higher cord blood IL-8, MPO and PGE(2) levels than full-term babies. These mediators, however, were not correlated to gestational age, suggesting that increases of these mediators are more related to preterm delivery but not fetal maturation. Further analysis showed that IL-8, a mediator mainly from monocytes, but not MPO, another mediator mostly from granulocytes, was correlated to cerebral palsy. CONCLUSIONS: Taken together, these results suggest that both premature monocytes and granulocytes activation are involved in preterm delivery, but maybe only monocytes activation is correlated to premature infants' cerebral palsy. Selective manipulation of monocytes activation may be useful to prevent premature-related cerebral palsy. PMID- 15113630 TI - Load perturbation does not influence spontaneous movements in 3-month-old infants. AB - BACKGROUND: The assessment of the quality of general movements (GMs) in young infants is a reliable and valid diagnostic tool for detecting brain dysfunction early in life. Of special interest is a type of GMs called fidgety movements (FMs) characteristic for 3- to 5-month-old infants. GMs are part of an infant's spontaneous motor repertoire and as such endogenously generated by the nervous system. Visual, acoustic and social stimuli hardly had any influence on FMs. AIM: Our main purpose was to find out whether FMs are sensitive to load perturbation. STUDY DESIGN: Spontaneous motility in supine position, with and without weighting was recorded on video and the data were semiquantitatively analysed. Weights were attached to the ankles and wrists of all four limbs; on one side of the body only; or without visual feedback of the weighted arm. SUBJECTS: We studied 29 healthy infants with normal FMs at the age of 12 weeks. RESULTS: Spontaneous motility remained symmetrical during all the experimental trails. Weighting had no influence on the quality or temporal organisation of FMs. CONCLUSION: This study demonstrated that the mechanisms responsible for FMs in 3-month-old infants are all but impervious to weight perturbation, at least not with the loads studied. FMs is the stable and predominant motor pattern of this age. PMID- 15113631 TI - Effects of estradiol administration on feto-placental growth in rat. AB - BACKGROUND: To clarify the effect of estradiol benzoate on placental structure and its consequences for fetal survival and fetoplacental growth. STUDY DESIGN: Estradiol benzoate (0, 0.1, 1, 10, 100 microg/day) was infused intraperitoneally into pregnant Wistar rats from 12 to 19 days' gestation. Survival rate, weight of pups and placentas at 20 days' gestation, and plasma levels of estrogen and progesterone were measured. Pathological changes in the placenta were also examined. RESULTS: Estradiol benzoate reduced fetal survival (1 microg/day: 100%, 10 microg/day: 70%, 100 microg/day: 14.6%) and the weights of the pups and placentas in a dose-dependent manner. Maternal estradiol concentration was raised 23-fold with 100microg/day of estradiol benzoate. Trophoblast degeneration, including apoptosis and destruction of placental labyrinth was induced but the structures of the maternal kidney and liver were not affected. CONCLUSIONS: In pregnant rats, estradiol benzoate causes fetal mortality at a pharmacological dose (more than 10 microg/day) and fetoplacental growth retardation via trophoblastic degeneration and destruction of the placental labyrinth even at a physiological dose (1 microg/day). PMID- 15113632 TI - Feeding and oral glucose--additive effects on pain reduction in newborns. AB - AIMS: The aims of this study were to compare the pain reducing effect of oral glucose with that of being breast-fed shortly before venipuncture in newborns, and also the pain score and crying time with parents' assessment. DESIGN: Randomised, controlled trial. SUBJECTS: 120 full term newborns undergoing venipuncture randomly assigned to on of four groups: I, Breast-fed and 1-ml placebo; II, Breast-fed and 1-ml 30% glucose; III, Fasting and 1-ml placebo; and IV, Fasting and 1-ml 30% glucose. OUTCOME MEASURES: Pain during venipuncture was measured with the Premature Infant Pain Profile (PIPP). Crying time was recorded. The parents assessed their babies' pain on a Visual Analogue Scale (VAS). RESULTS: The PIPP score was significantly lower in the infants receiving glucose, than in those not given glucose (p=0.004). There was no significant difference in PIPP score between the infants who were fed and the fasting infants. The PIPP score was lower in group II (median 7) than in group I (md 10). There was a similar difference between group IV (md 9) and group III (md 11). The median crying times during the first 3 min in groups I, II, III, and IV were 63, 18, 142 and 93 s, respectively. There was low agreement between the parents' assessment of pain and the PIPP score and crying time. CONCLUSION: Breast-feeding shortly before venipuncture has no major impact on the pain score but on crying time. The combination of oral glucose and breast-feeding shows the lowest pain score and significantly shorter duration of crying. PMID- 15113633 TI - Role of transforming growth factor-beta in breast milk for initiation of IgA production in newborn infants. AB - BACKGROUND: Transforming growth factor (TGF)-beta has a crucial effect on IgA production, which is the major humoral effector of mucosal immunity. Breast milk contains the abundant amount of TGF-beta in the early period of lactation. AIM STUDY DESIGN: To verify the notion that TGF-beta in breast milk might contribute to the development of IgA production in newborns, we investigated the association of TGF-beta in maternal colostrum with an increase of serum IgA in newborns during the first month of life. SUBJECTS AND METHODS: The concentrations of TGF beta1 and TGF-beta2, including IL-6 and IL-10, in colostrum samples from 55 healthy mothers were determined by ELISA. The levels of IgA and IgM in serum samples collected from corresponding newborn babies at birth and at 1 month of age were measured by ELISA. RESULTS: TGF-beta1 and TGF-beta2 were detected in substantial quantities in all colostrum samples, but IL-6 and IL-10 were present only in a proportion of samples. An increase of serum IgA in newborn during the first month of life was significantly higher than that of serum IgM (p<0.001). Notably, an increase of serum IgA in newborns during 1 month of life was well correlated with levels of both TGF-beta1 (r=0.38, p=0.005) and TGF-beta2 (r=0.45, p=0.0005) in colostrum, while that of IgM was marginally correlated with colostral TGF-beta2 (r=0.28, p=0.04). The association of increase of serum IgA in newborns with IL-6 and IL-10 in colostrum was not evident. CONCLUSION: Our findings suggest that TGF-beta in colostrum might serve as the starter of IgA production in newborn infants. PMID- 15113634 TI - Severe umbilical cord inflammation-a predictor of periventricular leukomalacia in very low birth weight infants. AB - Chorioamnionitis has been associated with periventricular leukomalacia (PVL) in very low birth weight (VLBW) infants. We examined the association between the pathological severity of chorioamnionitis and PVL in VLBW infants. Thirty-four VLBW infants with PVL and 34 control infants matched for gestational age without a diagnosis of PVL or intraventricular hemorrhage were obtained from the Women and Infants' Hospital of Rhode Island's Neonatal Follow-up Clinic database. Placental samples, including the amnion/chorion, chorionic plate, and umbilical cord, were examined microscopically. Statistical analysis included Mantel Haenszel chi-square, and Student's t-test. Severe inflammation in the umbilical cord was observed in 53% of infants with PVL and 32% without PVL (p<0.05). Severe umbilical cord inflammation is one of the risk factors associated with the development of PVL in VLBW infants. PMID- 15113635 TI - DNA damage in healthy term neonate. AB - BACKGROUND: The process of childbirth is accompanied by an increase in oxidative aggression. AIM: To determine DNA damage and oxidative stress in healthy term neonates at birth. DESIGN: A total of 34 healthy full-term neonates, 22 healthy adults and 20 samples of colostrum from mothers of full-term neonates were examined. The malondialdehyde (MDA), DNA damage, GSH/GSSG ratio and total antioxidant capacity (TAC) in mononuclear cells isolated from umbilical blood and adult peripheral blood were measured. Moreover, the TAC of colostrum was also measured. The protective activity of five natural polyphenols against H(2)O(2) induced DNA damage in mononuclear cells of umbilical blood was studied. RESULTS: A high level of DNA damage (p<0.001) accompanied with lower TAC (p<0.05) and GSH/GSSG ratio (p<0.001) and with higher level of MDA (p<0.001) in umbilical blood compared with those of healthy adult peripheral blood. The natural polyphenols, 7,8-dihydroxy-4-methyl coumarin, quercetin and resveratrol, are able to protect mononuclear cells of umbilical blood from oxidative attack. However, other two polyphenols, rutin and 7-hydroxy-4-methyl coumarin, do not. The TAC of colostrum is significantly higher than that of umbilical blood (p<0.001). CONCLUSIONS: The DNA oxidative damage in mononuclear cells of umbilical blood as well as other indexes related to redox status provided evidence that a sudden increase in oxygenation exposes the neonate into oxidative stress. Colostrum with a significant high TAC is very important for health care in infants against the oxidative stress. PMID- 15113636 TI - Pacifier use modifies infant's cardiac autonomic controls during sleep. AB - OBJECTIVE: The risk for sudden infant death (SIDS) was postulated to decrease with the use of a pacifier and by conditions increasing parasympathetic tonus during sleep. We evaluated the influence of a pacifier on cardiac autonomic controls in healthy infants. STUDY DESIGN: Thirty-four healthy infants were studied polygraphically during one night: 17 infants regularly used a pacifier during sleep and 17 never used a pacifier. Thumb users or occasional pacifier users were not included in the study. The infants were recorded at a median age of 10 weeks (range 6-18 weeks). Autonomic nervous system (ANS) was evaluated by spectral analysis of the heart rate (HR). The high frequency component of HR spectral analysis reflected parasympathetic tonus and the low frequency on high frequency ratio corresponded to the sympathovagal balance. RESULTS: Most infants (63.6%) lost their pacifier within 30 min of falling asleep. Sucking periods were associated with increases in cardiac sympathovagal balance. During non-sucking periods, in both REM and NREM sleep, infants using a pacifier were characterized by lower sympathetic activity and higher parasympathetic tonus compared with non pacifier users. CONCLUSIONS: The use of pacifiers modifies cardiac autonomic controls during both sucking and non-sucking sleep periods. Non-nutritive sucking could regulate autonomic control in infants. These findings could be relevant to mechanisms implicated in the occurrence of sudden infant deaths during sleep. PMID- 15113637 TI - The 2-week rule for patients with suspected breast cancer: what can be learnt by analysing policy documents? AB - The '2-week rule', introduced in the UK during 1999, represents a significant organisational change for referral of patients with suspected breast cancer. From an analysis of policy documents, a mixture of influences and agendas from different interest groups are apparent in the current working of the policy. These include political/modernisation agendas with a variety of aims including: reducing variation in care, efficient administration, reassuring patients, improving public confidence, earlier referral and lowering the threshold for referral to improve mortality figures. Specialist agendas, exemplified by the guidelines for referral under the policy but apparent in preceding specialist literature, represent an attempt to modify the working of the policy to ensure that high proportions of those women referred have breast cancer. The agendas of these interest groups are in conflict and have implications for the future development of this particular policy. Similar considerations are likely to apply more widely to other controversial health policy developments. It may be of value to identify the agendas of the groups responsible for the introduction of such policies, as well as the agendas of groups with influence on how the policy is actually implemented, and assess the areas of conflict. PMID- 15113638 TI - Lessons from Korea's pharmaceutical policy reform: the separation of medical institutions and pharmacies for outpatient care. AB - In Korea, until recently, both physicians and pharmacists were allowed to prescribe and dispense drugs for outpatient care. Along with other deep-rooted structural problems, this worked against the quality and efficiency of the health care system. To rectify this problem, the Korean government launched a drug policy reform in July 2000. However, the drug policy reform was more drastic than initially intended--driven by political factors, the reform ended up bringing about complete separation of medical institutions and pharmacies. Also, unlike in many other countries, Korea did not take a gradual approach, but instead, it implemented the reform all at once and nation-wide. As a result, the reform has faced criticism and protests, thereby generating unprecedented social turmoil and even strikes by physicians. Still, it is not clear what benefits Korea gained from this reform, when we look at the price which has had to be paid, including greater inconvenience, worsened access to medical care, increased drug spending, increased market share for multinational drug producers, and a greater deficit in the budget of the Korea's national health insurance system. Based on Korea's costly experience, we attempt to draw some policy implications for the future development of a better health care system. PMID- 15113639 TI - Changes in drug utilization following the outpatient prescription drug cost sharing program--evidence from Taiwan's elderly. AB - This paper examines changes in drug utilization following Taiwan's newly implemented National Health Insurance (NHI) outpatient prescription drug cost sharing program for persons over 65 years old. The study is a hospital outpatient prescription level analysis that adopts a pretest-posttest control group experiment design. Selected measures of outpatient prescription drug utilization are examined for cost-sharing and non cost-sharing groups in cost-sharing periods and pre cost-sharing periods. Additional analyses were conducted comparing older patients with and without chronic diseases and differences for essential and non essential drugs. Patients over age 65 were drawn from 21 hospitals in the Taipei area using a stratified random sampling method. This paper yields several interesting findings. First, average prescription cost and prescription period increased for both the cost-sharing and non cost-sharing groups. However, the rate of increase was significantly less in the cost-sharing group when compared with the non cost-sharing group. Second, the elderly with non-chronic diseases were more sensitive (i.e., reducing drug utilization) to the drug cost-sharing program when compared with those with chronic diseases. Third, for the elderly with non-chronic diseases average drug cost per prescription experienced a smaller decrease in essential drugs but a moderate increase in non-essential drugs for the cost-sharing group. By contrast, for the non cost-sharing group, average drug cost per prescription increased sharply in non-essential drugs as well as essential drugs. Finally, there was a significant increase in the number of prescriptions as well as drug costs above the upper bound of the cost-sharing schedule. The outpatient drug cost-sharing program implemented by the NHI in Taiwan did not reverse the trend of prescription drug cost increases in hospitals. The significant increase in the number of prescriptions above the upper bound of the cost-sharing schedule implies that the NHI should increase the upper bound. Further analysis needs to evaluate any adverse clinical impact for older patients resulting from policy changes. PMID- 15113640 TI - Comparing willingness-to-pay: bidding game format versus open-ended and payment scale formats. AB - The willingness-to-pay technique is being used increasingly in the economic evaluation of new health care technologies. Clinical trials of two methods of screening for colorectal cancer are currently being conducted in the UK and willingness-to-pay for screening has already been estimated by means of a questionnaire survey, using open-ended (OE) and payment scale (PS) formats. This paper addresses the same medical issue, although it elicits willingness-to-pay values by means of a bidding game in an interview setting. Interviews were conducted with 106 subjects in Nottingham. The bidding game format produced considerably higher valuations than had either of the previous questionnaire formats, whilst the significant differences between agreed valuations obtained using different initial bids supported the existence of starting-point bias in the bidding game. As with the questionnaire study, the majority of interview subjects offered relative valuations of tests at variance with their expressed preferences over the same tests. Given the significant difference in valuations generated by different formats, it follows that the economic case for preferring any one technology over others will depend considerably upon whichever format happens to have been used to generate the valuations. PMID- 15113641 TI - Seniors' prescription drug cost inflation and cost containment: evidence from British Columbia. AB - We develop an analytic framework to map out the nature and relative importance of different cost-driving trends in the prescription drug market. This is used to measure prescription drug cost-drivers for the population of seniors in British Columbia during a period when they received comprehensive public drug coverage. Between 1991 and 2001, expenditures on prescription drugs for BC seniors increased from dollar 149 to 320 million. Increases in the population of seniors, and the rate at which they utilized therapies contributed under half of the total cost increase over the period. Changes in the mix of therapies and the type of product selected explained over half of the observed drug expenditure inflation. Increased generic substitution significantly reduced the price of products selected over the period. PMID- 15113642 TI - The desire for support and respite care: preferences of Dutch informal caregivers. AB - Informal care is an indispensable element in the care for many patients. In order to maintain a sustainable input of informal care, it seems important to identify measures to alleviate the burden of care giving for caregivers at risk of burn out or other serious health problems, such as support and respite care. Thus, far research has focused on the burden of caregiving and on the supply of respite care. The demand side: what type of care is preferred by informal caregivers and what determines their preferences, is virtually unknown. We analysed the preferences and the underlying determinants for several types of support and respite care in a sample of 950 Dutch informal caregivers. Almost 80% of the respondents desire support or respite care in general, 42-47% would prefer more communication with other informal caregivers or more information of professional caregivers. Some time off is preferred by 40% of the respondents. The results show that caregiver characteristics, care recipient characteristics, elements of the caregiving situation and institutional variables determine the desire for support and respite care. Especially, the subjective burden of caregiving is important, whereas the number of caregiving tasks and the time invested (objective burden) hardly affect the desire for support and respite care. PMID- 15113643 TI - Public policy and smoking cessation among young adults in the United States. AB - In the wake of significant budget shortfalls, numerous states have increased cigarette excise taxes to boost revenues. This study examines whether or not increasing the price of cigarettes, which will occur as a consequence of cigarette excise tax increases, and implementing stronger restrictions on smoking in private worksites and other public places have an impact on smoking cessation decisions of young adults, thereby influencing public health in the United States (US). This paper employs longitudinal data on young adults from the Monitoring the Future Surveys matched with information on site-specific prices and measures of clean indoor air restrictions. A Cox regression is employed to estimate the smoking cessation equations. The estimates clearly indicate that increasing the price of cigarettes increases the number of young adults who quit smoking. The average price elasticity of cessation is 0.35. In addition, stronger restrictions on smoking in private worksites and public places other than restaurants increase the probability of young adult smoking cessation. Given the well-documented benefits of smoking cessation, a significant increase in cigarette excises taxes may be one of the most effective means to reduce premature death and disease in the United States. PMID- 15113644 TI - A large proportion of Swedish citizens refrain from seeking medical care--lack of confidence in the medical services a plausible explanation? AB - Equity in health and health care is and has been a long-standing goal in Swedish health care politics. This study aims to look into how different socio demographic variables influence unmet needs i.e. why one would refrain from seeing a doctor, despite a perceived need for medical care. A nation-wide postal questionnaire was answered by 2648 (66%) randomly chosen individuals in the ages between 20 and 64 years. The questionnaire included questions on health and health care utilisation along with data on different socio-demographic variables. The proportion of citizens that refrain from visiting a physician despite a perceived need was higher (24%) than in any previous Swedish investigation. Women, those of a non-Swedish origin and those with a low level of education refrained from going to the physician to a higher extent than men, inborn citizens and those with a higher education. Stated reasons to why the respondents refrained from medical care were associated with confidence, primarily, finite availability and economy. It appears as the Swedish health care system is not fully adapted to provide for the so far unmet needs of a large proportion of the population and that this has equitable concerns. PMID- 15113645 TI - The predictability of individual primary care costs and its impact on managed care plans. AB - In the context of reorganizing the delivery of primary care, a better understanding of the evolution of family practice costs would be useful for those managing the health care system. This study examined the level of use and transition probabilities of remaining a low user of physician care in family practices over 5 years among a population with no restriction of age or functional status. The entire population (n = 285, 305) of one administrative area of the Province of Quebec (Canada) was followed from 1996 to 2000 and classified for each year into seven cost categories according to individual costs of care in family practices. The Pr of staying in the same category or moving to another one has been examined and these transitions were calculated for 2, 3, 4 and 5 year-periods. Recourse to care was very stable over time. The Pr of staying in the same cost category or moving to the adjacent one was always 91% between 2 years and still as high as 86% between 4 years. Less than 5% of the population jumped over more than two categories. More than three quarters of the population never exceeded Candollar 200 over the 5 years while only 2.5% had a chance to reach Candollar 500. Costs of care in family practices were highly correlated with visits (r = 0.94), suggesting the role of chronicity as a determinant of expenditures in these settings. The stability of individual costs of care in family practices illustrated by the high transition probabilities over time indicates the possibility to forecast these costs in a perspective of capitation payment plans. PMID- 15113646 TI - Impact of national health care systems on patient evaluations of general practice in Europe. AB - OBJECTIVE: To examine associations between patient evaluations of general practice and characteristics of national health care systems. METHODS: International comparative study in 17 countries, using international patient survey data (n= 25052) and data-bases for health care system characteristics. Dependent variable was patients' evaluation of general practice care. Five independent factors were examined: GP density, physician density, fee for service reimbursement, gatekeeper role, and first-contact role. RESULTS: None of the associations were statistically significant. CONCLUSIONS: Patients were highly satisfied with general practice in different national health care systems. Descriptive tables on international variations may lead to misleading conclusions. PMID- 15113647 TI - Managing under managed community care: the experiences of clients, providers and managers in Ontario's competitive home care sector. AB - In 1996, a newly elected government in the Province of Ontario, Canada, introduced a managed competition environment into the home care sector through the establishment of a competitive contracting process for home care services. Through 65 in-depth, semi-structured interviews conducted between November 1999 and January 2001, we trace the implementation of this competitive contracting policy within Ontario's newly established managed community care environment and assess the effects of competitive contracting against two sets of goals: (1). quality of care goals that consider continuity of care of paramount importance in the provision of home care; and (2). the managed competition goal of increased efficiency. In assessing the implementation of this policy against these goals, we highlight the conflicts that can arise in pursuing different policy goals in response to different formulations of the policy problem that underpin them. We map stakeholder experiences with the competitive contracting policy onto relevant contracting and managed competition literatures. When measured against the goals of quality of care and efficiency, the findings presented here offer a mixed review of the experiences to date with the competitive contracting process introduced in Ontario's home care sector and suggest improvements for managing future competitive contracting processes. PMID- 15113648 TI - Reserved procedures in Dutch hospitals: knowledge, experiences and views of physicians and nurses. AB - The Individual Health Care Professions Act came into force in The Netherlands in 1997, introducing a mixed system for the regulation of the practice of medicine. One of its components, the reserved procedures regulations, was studied in hospitals to gain insight into the knowledge, experiences and views of physicians and nurses with regard to these regulations. Questionnaires were sent to representative samples of 250 gynaecologists, 350 internists, and 3200 nurses, response rates were 65, 60 and 71%, respectively. Almost all respondents were aware that physicians are authorised to perform reserved procedures on their own initiative (93-99%), and 48-63% knew that nurses are not authorised to do this. A substantial percentage of the nurses performed reserved procedures on their own initiative (17-53%). A majority of gynaecologists and internists presumed that the hospital had ensured the proficiency of the nurses to perform reserved procedures (58% resp. 65%), while 82% of the nurses determined their own proficiency for each procedure. Most respondents felt that the reserved procedures regulations offer adequate protection for patients (58-72%). Although recommendations are made for improvement, the functioning of the reserved procedures regulations in hospitals is considered to be moderately positive. PMID- 15113649 TI - Cloning and sequence analysis of llama cytokines related to cell-mediated immunity. AB - In order to characterize the T helper 1 (Th1) cytokines of llama, we have cloned several llama cytokine genes and compared them to those of other mammalian species. The cDNAs encoding for interleukin (IL)-2, interferon (IFN)gamma, IL 12p35 and IL-12p40 were amplified using specific primers designed from reported sequences of bovine cytokine genes. The cDNAs for llama IL-2, IFN-gamma, IL-12 p35 and IL-12p40 were found to be 465, 501, 669 or 993 bp in length, with open reading frames encoding 154, 166, 222 or 330 amino acids, respectively. Homology analyses of nucleotide and deduced amino sequences of llama IL-2, IFN-gamma, IL 12p35 and IL-12p40 and phylogenetic analysis based on their nucleotide sequences indicated the close relationship in these cytokine genes between llama and eutherian mammalian order Artiodactyla, which includes pig and cattle. PMID- 15113650 TI - Recombinant bacteriophage-based multiepitope vaccine against Taenia solium pig cysticercosis. AB - The aim of this study was to test the capacity of recombinant phages to deliver antigens for vaccination against porcine cysticercosis. Thus, three peptides (KETc1, KETc12, GK1) and a recombinant antigen KETc7, previously proven to induce high levels of protection against pig cysticercosis, were expressed on the surface of the M13 bacteriophage at multiple copies. The pool of these four recombinant phages induced high levels of protection against an experimental murine cysticercosis. The immunogenicity of the phage vaccine preparation was therefore, tested in pigs, the natural host of Taenia solium. Subcutaneous or oral vaccination with these phages induced antigen-specific cellular immune responses in pigs. Preliminary data also points to the protective capacity of this recombinant phage vaccine against pig cysticercosis. The immunogenicity of these recombinant phages, together with the low cost of their production, make them a realistic candidate to be tested in pigs as an anti-cysticercus phage vaccine for field trials. This is the first report describing the application of a filamentous bacteriophage as a vaccine in large animals such as pigs, the only intermediate hosts of T. solium, a parasite of major medical importance in developing countries. The potential application of phages as a modern platform for vaccines for human and animal diseases is discussed. PMID- 15113651 TI - Spontaneous T cell apoptosis in feline immunodeficiency virus (FIV)-infected cats is inhibited by IL2 and anti-B7.1 antibodies. AB - Lymph node (LN) T cells from feline immunodeficiency virus (FIV)-infected cats have an increased expression of B7 co-stimulatory molecules as well as their ligand CTLA4, resembling an activation phenotype shown to induce anergy and apoptosis in activated T cells. In addition, LN T cells from FIV-infected cats also show increased spontaneous apoptosis compared to uninfected animals. The apoptosis observed in these animals occurs primarily in T cells expressing B7 and CTLA4, suggesting a role for B7 and CTLA4 interactions in the induction of anergy/apoptosis. In order to investigate the role of B7 and CTLA4 interactions on T cell apoptosis in LN T cells from FIV-infected cats, we performed blocking experiments by measuring T cell apoptosis in LN T cell cultures treated with anti feline B7.1, B7.2, and CTLA4 specific antibodies, as well as interleukin (IL)-2. The addition of IL2, the primary cytokine produced by B7/CD28 interactions, resulted in a significant decrease of T cell apoptosis in cultured LN cells as assessed by two-color flow cytometry and TUNEL assay. The addition of anti-B7.1 antibodies significantly inhibited T cell apoptosis in FIV-infected cats with low level plasma viremia, while addition of anti-B7.2 and anti-CTLA4 antibodies had no affect. These results suggest a role of B7 signaling in the increased spontaneous apoptosis observed in LN T cells from FIV-infected animals. PMID- 15113652 TI - Upregulation of transforming growth factor-beta and interleukin-10 in cows with clinical Johne's disease. AB - Johne's disease progresses through distinct stages including a protracted subclinical stage in which the infection appears to be controlled; followed by a more acute stage in which the host animal demonstrates clinical signs such as diarrhea and weight loss. Little is known about the dynamics of the host immune response during these two phases of disease, however, it is possible that immune modulation in the early stages of disease may play an important role in disease progression. We hypothesized that the clinical stage of Johne's disease is mediated by the expression of cytokines such as transforming growth factor-beta (TGF-beta) and interleukin-10 (IL-10) that may be accompanied by the downregulation of IFN-gamma gene expression. In the present study, tissue samples were collected from the ileum, ileocecal junction, ileocecal lymph node, and mesenteric lymph nodes of healthy, subclinically or clinically infected cows. The expression of TGF-beta, IL-10, and IFN-gamma genes in these tissues was determined by quantitative competitive RT-PCR. The results demonstrate that TGF beta and IL-10 mRNA levels are higher in cows that have progressed to the clinical stage of disease compared to subclinically infected or healthy cows. In contrast, IFN-gamma gene expression was significantly higher in subclinically infected cows. These results suggest that a change in the balance of cytokines at the site of infection may contribute to the ability of the host to control Mycobacterium avium subsp. paratuberculosis infection. PMID- 15113653 TI - Selection of HPRT primers as controls for determination of mRNA expression in dogs by RT-PCR. AB - Reliable housekeeping gene controls are critical for measuring and comparing gene expression at the transcription level by Northern blot and RT-PCR. In order to develop such controls for studying cytokine mRNA expression in dogs, DNA sequence encoding a full-length canine HPRT protein has been obtained. Numerous primer pairs derived from the canine HPRT sequence have been tested on canine genomic DNA as well as cDNA. The data from the present study suggest that there may be processed HPRT pseudogenes in dogs. Three pairs of canine HPRT primers designed and tested in the present study were able to differentiate between cDNA and genomic DNA under specific PCR conditions. These primers would be useful controls for measurement of mRNA expression by RT-PCR in the dog. PMID- 15113654 TI - Hematopoiesis in the feline fetal liver: an assessment by flow cytometry. AB - The domestic cat is an excellent model for the development of therapeutic protocols that target hematopoietic stem cells (HSCs) because it is relatively resistant to complications related to bone marrow transplantation. To identify a plentiful source of HSC that could be used as targets for gene transduction and transplantation, the livers of 28 mid-gestation fetuses (28-52 days) and late gestation fetuses (53 days-term) were analyzed for erythroid, myeloid, lymphoid, and uncommitted hematopoietic progenitor cells by flow cytometry. We found that the fetal liver mononuclear cells (FLMCs) contained 57% erythroid progenitors during mid-gestation, but this percentage declined to 43% as gestation progressed. Myelomonocytic cells within FLMC were more numerous in late-gestation (31%) than in mid-gestation (18%). Two monoclonal antibodies (mAb), CH 152 and CH 755, which recognize cells with the potential to reconstitute multilineage hematopoiesis in cats, were tested. Approximately, 32% of FLMC from late gestation fetuses expressed the epitope recognized by mAb CH 152, a significant increase above the 12% positive cells in mid-gestation fetuses. Approximately, 33% of hepatic mononuclear cells expressed the epitope recognized by mAb CH 755 in both mid-term and late-term fetuses. When expressed in absolute numbers, medians of 2.7 x 10(7) CH 152-positive cells and 3.2 x 10(7) CH 755-positive cells were extracted from the late-term fetal livers of individual cats. T lymphocytes were a minor component (<3%) of FLMC, despite their presence in the thymus and spleen. These data suggest that the late-term feline fetal liver is a suitable source of mutipotential hematopoietic cells that could be used for gene therapy protocols in the cat. PMID- 15113655 TI - Systemic cytokine profile in feeder pigs suffering from natural postweaning multisystemic wasting syndrome (PMWS) as determined by semiquantitative RT-PCR and flow cytometric intracellular cytokine detection. AB - Postweaning multisystemic wasting syndrome (PMWS) is an economically important disease in pigs caused by porcine circovirus type 2 (PCV2). Development of this disease is presumably associated with an impairment of the immune system. We, therefore, investigated the systemic expression of relevant cytokines (IL-1, IL 2, IL-4, IL-6, IL-8, IL-10, IL-12p40, TNF-alpha, IFN-gamma) and IL-2Ralpha at mRNA (semiquantitative RT-PCR) and at protein level (flow cytometric intracellular cytokine detection after short-time stimulation of peripheral blood mononuclear cells) in 10 feeder pigs aged 14 weeks suffering from natural PMWS and in 10 clinically healthy pen-mates. Hematological examination revealed a significant (p < 0.001) relative lymphopenia in the diseased animals when compared to reference pigs. IL-1alpha and IL-10 mRNA levels were notably increased in the affected pigs, whereas IL-2 and IL-2Ralpha (CD25) mRNA levels tended to be down-regulated. IL-8, TNF-alpha and IFN-gamma mRNA expressions appeared to be slightly increased. Intracellular cytokine levels as measured by flow cytometry revealed an increase of IL-1beta, IL-2, and IL-6, whereas IL-12 and TNF-alpha expressions were not affected. IFN-gamma was slightly decreased in the diseased animals. In conclusion, despite the assumption, that the cellular immune response to PMWS as a virus-induced disease should be characterized by either a Th1 driven cytokine profile or a cytokine profile indicative of T cell immunosuppression, our results did not support that hypothesis. Nevertheless, data from intracellular cytokine detection suggest an even increased percentage of the remaining lymphocytes capable to produce IL-2 upon in vitro stimulation, which is in contrast to the slightly diminished IL-2 mRNA levels reflecting the in vivo situation at least at the mRNA level. PMID- 15113656 TI - Characterization of canine microglial cells isolated ex vivo. AB - Microglia are the principal immune effector elements of the brain sharing immunophenotypic and functional characteristics of macrophages as well as of antigen presenting cells (APCs). The purpose of this study was to isolate canine microglial cells and make them available for ex vivo characterizations of their functions and immunophenotype. After isolation, carried out by density gradient centrifugation, microglial cells accumulated on distinct interfaces of 1.077 and 1.066 g/ml of a Percoll gradient. Identification of microglial cells in other species is realized by their specific immunophenotype of CD11b/c+ and CD45low. Our results indicate, that expression of CD45 is very low or even absent in canine microglial cells. In addition, they expressed CD18 and CD11b/c+, as determined by flow cytometry and immunohistochemistry. Fourteen additional monoclonal antibodies (mAbs) were used to characterize and compare canine microglial cells with monocytes. Microglia and monocytes can be clearly distinguished by their differential expression intensity of surface antigens (CD45, CD44, CD14). Functional characterization was assessed by a reactive oxygen species (ROS)-generation test and phagocytosis assay using flow cytometry. In conclusion, ex vivo examination of microglia is possible in dogs and most probably reflects the conditions in vivo. The measurement of tissue culture artifacts can be largely avoided using this method. PMID- 15113657 TI - Quantitative analysis of pro-inflammatory cytokine mRNA expression in Theileria annulata-infected cell lines derived from resistant and susceptible cattle. AB - The pathogenic mechanisms involved in tropical theileriosis, caused by the tick borne protozoan parasite Theileria annulata, are unclear. Pathology is associated with the schizont stage of the parasite, which resides within bovine macrophages. Breed-specific differences in pathology have been observed in cattle, several Bos indicus breeds are relatively resistant to tropical theileriosis whilst Bos taurus cattle are highly susceptible. Infected cells express pro-inflammatory cytokines and it has been hypothesized that these cytokines play a major role in the pathology of the disease. Therefore, using quantitative RT-PCR we investigated the expression of the key candidates, interleukin 1 beta (IL-1beta), IL-6 and tumour necrosis factor alpha (TNF-alpha), in T. annulata low passage infected cell lines derived ex vivo from experimental infection of resistant and susceptible cattle. mRNA for each cytokine was detected in all cell lines investigated at levels higher than those observed in resting monocytes. However, the analyses did not identify any breed-specific differences. Therefore, these results are not consistent with the hypothesis that differential regulation of infected cell derived pro-inflammatory cytokines (IL-1beta, IL-6 and TNF-alpha) accounts for the breed-related differences in resistance and susceptibility to T. annulata infection. Other, currently unknown mechanisms may be of greater importance. PMID- 15113658 TI - Cloning and sequencing of a cDNA expressing a ribosomal P0 peptide from Culicoides nubeculosus (Diptera). AB - Insect bite dermal hypersensitivity (IBH) is an allergic dermatitis of horses caused by bites of Culicoides spp. and sometimes Simulium spp. The aim of the investigation presented here was to identify allergens causing IBH. A cDNA library expressing recombinant Culicoides nubeculosus proteins was screened using affinity-purified serum from an IBH-affected horse. Screening of the library resulted in identification of one immunoreactive clone. The sequence of the cDNA insert was determined and revealed a 600 bp insert with an open reading frame coding for a 78 amino acid long protein, called rCul n 1. Analysis of the deduced amino acid sequence revealed an identity of 67-78% to the C-terminal part of the 318 amino acid long ribosomal P0 protein from other Diptera. Furthermore, the 38 C-terminal amino acids displayed an identity of 57% with the C-terminal part of the acidic ribosomal protein P2 from Aspergillus fumigatus. The cDNA insert was subcloned and expressed as a [His]6-tagged protein in Escherichia coli and purified using Ni2(+)-chelate affinity chromatography. The 10kDa recombinant Cul n 1 protein bound the affinity-purified antibody fraction used for screening the expression library. Determination of IgE and IgG levels against rCul n 1 by ELISA in sera from 19 IBH-affected and 18 Swiss control horses and in sera from eight control horses living in Iceland showed no significant differences between the three groups of horses (median IgE levels = 60, 49 and 44 relative ELISA units, respectively). rCul n 1 did not induce sulfidoleukotriene (sLT) release from peripheral blood leukocytes of IBH-affected horses (N = 5), although sLT release was induced with the Culicoides whole body extract. PMID- 15113659 TI - Vaccine-induced T cell-mediated immunity plays a critical role in early protection against pseudorabies virus (suid herpes virus type 1) infection in pigs. AB - The aim of our study was to evaluate the relative importance of antibody and T cell-mediated immunity in protection against pseudorabies virus (suid herpes virus type 1) infection in pigs. We induced different levels of immune responses by using: (1) a modified live vaccine; (2) the same modified live vaccine with an oil-in-water (o/w) adjuvant; (3) an inactivated vaccine; and (4) the same inactivated vaccine with an o/w adjuvant. Subsequently, we challenged pigs with virulent pseudorabies virus (PRV). We demonstrated that best-protected pigs stood out by maintaining strong T cell-mediated immune (CMI) responses after challenge. Of the immune parameters tested, protection against virus shedding was correlated best with the magnitude of the IFN-gamma response of in vitro re-stimulated peripheral blood mononuclear cells (PBMC) with an additional role for PRV specific IgG2 antibodies. The use of an o/w adjuvant resulted in higher antibody and CMI responses, in particular with an increased frequency of memory T helper blast cells of in vitro re-stimulated PBMC. However, this adjuvant-induced enhancement of the immune response had a limited additional effect on the efficacy of inactivated vaccines. This study suggests a major contribution of the CMI response in early protection against PRV infection and that PRV-induced IFN gamma responses may serve as a suitable indicator for assessing the immune status of vaccinated pigs. PMID- 15113660 TI - Equine beta-defensin-1: full-length cDNA sequence and tissue expression. AB - beta-Defensins are cysteine-rich endogenously produced antimicrobial peptides that play an important role in innate immune defense. Although, previous investigations have identified beta-defensins in several mammalian species, no reports have identified equine beta-defensins. Using a strategy of database searching for expressed sequence tags (EST) we identified putative expression of equine beta-defensins in hepatic tissue. Based on this information, sequence specific primers were designed for the equine gene enabling the identification of the full-length cDNA sequence of equine beta-defensin-1. Comparative analyses showed that equine beta-defensin-1 has 46-52% amino-acid identity with other beta defensins, sharing the greatest identity with porcine beta-defensin-1. Complete conservation of cysteine residues was maintained between the species evaluated, and RT-PCR analysis revealed diverse mRNA tissue expression for equine beta defensin-1. These data extend the repertoire of equine antimicrobial peptides and expand our understanding of equine innate immunity. PMID- 15113661 TI - Synthesis and properties of new bolaform and macrocyclic galactose-based surfactants obtained by olefin metathesis. AB - A series of galactose-based surfactants with various structures likely to display new interesting properties were synthesized. Four monocatenary surfactants were elaborated by microwave-assisted galactosylation of undecanol or 10-undecenol. These compounds were slightly soluble in water. Their tensioactive properties were determined at 45 degrees C. Olefin metathesis was used to synthesize the two single-chain bolaforms from undec-10-enyl galactopyranosides; two pseudomacrocyclic bolaforms were prepared by grafting two carbamates at O-4 and O 4' sugar positions of the single-chain bolaforms. These four surfactants are insoluble in water and undergo monolayer compression. Cyclization of these bolaforms by olefin metathesis led to macrocyclic surfactant analogues of archaeobacterial membrane components. PMID- 15113662 TI - Gram-scale syntheses of the (1-->3)-linked and (1-->4)-linked hyaluronan disaccharides. AB - The first gram-scale syntheses of two hyaluronan disaccharides are described. Construction of the (1-->4)-linked disaccharide 12 was achieved in 12% overall yield using 2,3-bis-dimethyl acetal protection in combination with chlorosilane induced carbamate cleavage methodologies. The uronic acid functionality was installed using TEMPO oxidation with NaOCl as the hypochlorite source. The (1- >3)-linked disaccharide 18 was achieved in 7% overall yield utilizing acetonide protection in addition to the chlorosilane-induced carbamate cleavage methodology and the TEMPO oxidation. PMID- 15113663 TI - The synthesis of some deoxygenated analogues of early intermediates in the biosynthesis of glycosylphosphatidylinositol (GPI) membrane anchors. AB - Syntheses are described of 2-azido-4,6-di-O-benzyl-2,3-dideoxy-d-ribo hexopyranosyl fluoride, 6-O-acetyl-2-azido-3-O-benzyl-2,4-dideoxy-d-xylo hexopyranosyl fluoride and 2-azido-3,4-di-O-benzyl-2,6-dideoxy-d-glucopyranosyl fluoride. These glycosyl donors were coupled with the acceptor 1d-2,3,4,5-tetra-O benzyl-1-O-(4-methoxybenzyl)-myo-inositol and the alpha-coupled products were transformed into alpha-d-3dGlcpN-PI, alpha-d-4dGlcpN-PI and alpha-d-6dGlcpN-PI by way of the H-phosphonate route. Brief mention is made of the biological evaluation of these deoxy-sugar analogues and their N-acetylated forms as candidate substrate/inhibitors of the N-deacetylase and alpha-(1-->4)-d mannosyltransferase activities present in trypanosomal and HeLa (human) cell-free system. PMID- 15113664 TI - Regioselective monoacylation of cyclomaltoheptaose at the C-2 secondary hydroxyl groups by the alkaline protease from Bacillus subtilis in nonaqueous media. AB - Transesterification of cyclomaltoheptaose (beta-CD) with divinyl butanedioate, divinyl hexanedioate, and divinyl decanedioate, catalyzed by the alkaline protease from Bacillus subtilis in anhydrous DMF for 5 days, furnished the corresponding vinyl-beta-CD derivatives. The products were characterized by ESI MS, (1)H NMR, (13)C NMR, IR, and DSC. The results indicated the products to be monosubstituted esters, with monoacylation occurring at the C-2 secondary hydroxyl groups of beta-CD. The regioselectivity of the monoacylation as catalyzed by alkaline protease was not affected by the chain length of the acyl donor. PMID- 15113665 TI - Synthesis of L-arabinose-containing fragments of the oat root saponin Avenacin A 1. AB - Chemical syntheses of two disaccharides, benzyl beta-D-glucopyranosyl-(1-->2) alpha-L-arabinopyranoside (1) and benzyl beta-D-glucopyranosyl-(1-->4)-alpha-L arabinopyranoside (2), and a trisaccharide, benzyl beta-D-glucopyranosyl-(1-->2) 3-O-acetyl-4-O-(beta-D-glucopyranosyl)-alpha-L-arabinopyranoside (3), related to oat root triterpenoid saponin Avenacin A-1 are reported. PMID- 15113666 TI - Synthesis of cis-(1-->3)-glycosides of allyl 2-acetamido-4,6-O-benzylidene-2 deoxy-alpha-D-glucopyranoside. AB - Syntheses of allyl 2,3,4-tri-O-benzyl-alpha-D-gluco- and D galactopyranosyluronate-(1-->3)-2-acetamido-4,6-O-benzylidene-2-deoxy-alpha-D glucopyranoside via oxidation of the hydroxymethyl group of allyl 2,3,4-tri-O benzyl-alpha-D-gluco- and D-galactopyranosyl-(1-->3)-2-acetamido-4,6-O benzylidene-2-deoxy-alpha-D-glucopyranoside under Jones conditions are described. Structures of the title compounds were confirmed by (1)H and (13)C NMR spectroscopy. PMID- 15113667 TI - Isolation of kappa-carrageenan oligosaccharides using ion-pair liquid chromatography--characterisation by electrospray ionisation mass spectrometry in positive-ion mode. AB - Oligo-kappa-carrageenans participate as elicitors in the cell-cell recognition process in marine plants. Analytical methods can be usefully applied to gain insight into the biochemistry of these biological processes. Therefore, enzymatically digested oligomers of kappa-carrageenans have been separated and isolated on a Spherisorb ODS1 (250 x 4 mm i.d., particle size 5 microm) column using ion-pair liquid chromatography coupled with an evaporative light scattering detector. Heptylamine (5 mM, pH4) has been selected as the ion-pairing agent and MeOH as the organic modifier in a gradient mode. Overloading the column with 1mg of the mixture, the chromatographic mechanism presented adequate stability. The mobile phase of each isolated oligomer was evaporated and the residue was infused into an electrospray ionisation mass spectrometry (ESIMS) in positive-ion mode with 4:1 MeCN-water as mobile phase. Each ESIMS spectrum presented ions consisting of the oligomer attached with a number of heptylammonium ions depending on the molecule size. In addition, the different m/z values permitted direct detection of the oligomers in ESIMS positive-ion mode. The analytical method developed separated the oligomers up to dotriacontasaccharide. PMID- 15113668 TI - Synthesis, self-assembly and characterization of a new glucoside-type hydrogel having a Schiff base on the aglycon. AB - A new hydrogel based on a substituted phenyl glucoside with a Schiff base in the aglycon was synthesized, and the self-assembling characteristics was studied. FTIR spectra, UV-vis absorption spectra and X-ray diffraction (XRD) revealed that pi-pi interactions between the Schiff base moieties, hydrogen bonds, and the interdigitated interactions between hydrophobic chains had effects on the formation of the self-assembling hydrogel. Scanning electron microscopic (SEM) and transmission electron microscopic (TEM) observation showed that the three dimensional hydrogel network was constructed from nanotubes with inner diameters of ca. 75 nm and wall of ca. 20 nm. PMID- 15113669 TI - Material properties of concentrated pectin networks. AB - We have examined the mechanical behaviour of different types of pectin at high concentrations (> 30% w/w), relevant to the behaviour of pectin in the plant cell wall, and as a film-forming agent. Mechanical properties were examined as a function of counterion type (K(+), Ca(2+), Mg(2+)), concentration and extent of hydration. Hydration was controlled in an osmotic stress experiment where pectin films were exposed to concentrated polyethylene glycol [PEG] solutions of known osmotic pressure. We investigated the mechanical behaviour under simple extension. The results show that the swelling and stiffness of the films are strongly dependent on pectin source and ionic environment. At a fixed osmotic stress, both Ca(2+) or Mg(2+) counterions reduce swelling and increase the stiffness of the film. PMID- 15113670 TI - A theoretical study of the conformational behavior of analogues of alpha-L rhamnose-1-phosphate. AB - The conformational behavior of methyl(2-O-methyl-alpha-L rhamnopyranosyl)phosphate, together with a group of potentially more stable analogues, was investigated through a DFT approach at the B3LYP/6-31G(d) level; the energy of all the optimized structures was recalculated using a continuum solvent model, C-PCM, choosing water as the solvent. The compounds exhibited several, sometimes tenths of populated conformations so that the overall properties of flexibility and mobility were evaluated. The analogue in which the pyranose oxygen atom is replaced by a methylene group emerges as the best candidate as a mimic of the reference 1-phosphate, in spite of the fact that it lacks the anomeric and exo-anomeric effects. The other analogues result poorer mimics because of a conformational equilibrium at the pyranose ring or of an excessive rigidity of the aglycone moiety. PMID- 15113671 TI - A conformational study of alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->O)-L-Ser by NMR 1H,1H T-ROESY experiments and molecular-dynamics simulations. AB - The conformational preference of alpha-D-Manp-(1-->2)-alpha-D-Manp-(1-->O)-L-Ser has been investigated by one-dimensional (1)H,(1)H T-ROESY experiments and molecular-dynamics simulations with CHARMM22 type of force fields and water as explicit solvent. Proton-proton distances were obtained from the simulations and subsequently experimentally determined distances could be derived. Measurements were performed on the title compound as well as on selectively deuterium substituted analogues synthesized as part of this study to alleviate possible NMR spectroscopic difficulties. A very good agreement was present between the separate NMR experiments. In the subsequent analysis a key nuclear Overhauser effect between the anomeric protons in the two sugar residues was used to assess the conformational dynamics revealed by the molecular simulations. The combined results support a model in which two states are significantly populated as a result of flexibility around the bond defined by the glycosidic torsion angle psi. PMID- 15113672 TI - Isolation and partial characterization of fucan sulfates from the body wall of sea cucumber Stichopus japonicus and their ability to inhibit osteoclastogenesis. AB - Two types of fucan sulfate were isolated from chloroform/methanol extract of the body wall of the sea cucumber Stichopus japonicus. One type (type A) contained 3.41 mmol fucose/g and 2.35 mmol sulfate/g, and the molecular mass was determined to be 9 kDa by gel permeation chromatography (GPC). Structural analysis suggested that type A consists of a backbone of (1-->3)-linked fucosyl residues that are substituted at C-4 with fucosyl residues, and that fucosyl residues are sulfated at C-2 and/or C-4. Another type (type B) contained 3.90 mmol fucose/g and 3.07 mmol sulfate/g, and the molecular mass was determined to be 32kDa by GPC. Structural analysis showed that type B is largely composed of unbranched (1-->3) linked fucosyl residues, and that sulfate substitution(s) occur at C-2 and/or C 4. The potential of both types to inhibit osteoclastogenesis was examined by an in vitro assay system, showing that both types of fucan sulfate inhibit osteoclastogenesis more than 95% at 50 microg/mL concentration. These results suggest that types A and B fucan sulfate from sea cucumber are potent inhibitors of osteoclastogenesis. PMID- 15113673 TI - Structure of a highly phosphorylated O-polysaccharide of Proteus mirabilis O41. AB - A highly phosphorylated O-polysaccharide was obtained by mild acid degradation of the lipopolysaccharide of Proteus mirabilis O41 followed by GPC. The initial and dephosphorylated polysaccharides and phosphorylated products from two sequential Smith degradations were studied by (1)H, (13)C and (31)P NMR spectroscopy and ESI MS. The O-polysaccharide was found to have a tetrasaccharide repeating unit containing one ribitol phosphate (presumably d-Rib-ol-5-P) and two ethanolamine phosphate (Etn-P) groups, one of which is present in the stoichiometric amount and the other in a nonstoichiometric amount. The following structure of the O polysaccharide was established: PMID- 15113674 TI - Lipase-catalysed preparation of acetates of 4-nitrophenyl beta-D-xylopyranoside and their use in kinetic studies of acetyl migration. AB - Di-O-acetates and mono-O-acetates of 4-nitrophenyl beta-D-xylopyranoside were prepared by use of lipase PS-30. Polarity of organic solvents and reaction time affected the regioselectivity of the di-O-acetylation as well as the yields of monoacetates. The kinetics of acetyl groups migration in these derivatives was studied in aqueous media using HPLC. Migration of the acetyl group strongly depended on pH. The highest rate of acetyl migration was observed from O-2 to O-3 in both 2,4-di-O-acetate and 2-O-acetate. On the contrary, acetyl exchange between O-3 and O-4 in both directions was slower than between O-2 and O-3. The 2,3-di-O-acetate and 4-O-acetate showed to be the most stable towards acetyl migration. The 3,4-di-O-acetate and 4-O-acetate were dominant in the corresponding equilibration mixtures. PMID- 15113675 TI - Synthesis of 6I-amino-6I-deoxy-2I-VII,3I-VII-tetradeca-O-methyl cyclomaltoheptaose. AB - The preparation of 6(I)-amino-6(I)-deoxy-2(I-VII),3(I-VII)-tetradeca-O-methyl cyclomaltoheptaose is reported. Two different routes (A and B), both starting from beta-cyclodextrin (betaCD), have been examined. Route A involved: (i) synthesis of heptakis(6-O-tert-butyldimethylsilyl)-betaCD from betaCD; (ii) permethylation of the secondary hydroxyl groups with methyl iodide and sodium hydride; (iii) desilylation of the primary hydroxyls with ammonium fluoride; (iv) monotosylation at O-6 position of per-(2,3-O-methyl)-betaCD; (5) nucleophilic replacement of the tosyl group with azide anion; (v) reduction of the azido group by catalytic transfer hydrogenation using hydrazine hydrate in the presence of Pd/C in methanol/water. Route B started from the known 6(I)-monoazido-6(I) monodeoxy-beta-CD (two steps from beta-CD) and entailed: (i) protection of the remaining primary hydroxyls using tert-butyldimethylsilylchloride (TBDMSCl); (ii) exhaustive methylation of the secondary hydroxyls with methyl iodide and sodium hydride; (iii) removal of the TBDMS protecting groups with ammonium fluoride; (iv) reduction of the azido group as above. Route A was found to be less convenient than Route B due to the inherent difficulty of controlling the monotosylation of per-(2,3-O-methyl)-betaCD. PMID- 15113676 TI - 2beta,3beta-Difluorosialic acid derivatives structurally modified at the C-4 position: synthesis and biological evaluation as inhibitors of human parainfluenza virus type 1. AB - A series of 4-O-substituted 2beta,3beta-difluorosialic acid derivatives (3a-d) has been synthesized. A key intermediate was synthesized efficiently by the electrophilic syn-addition of fluorine to the double bond of a glycal precursor using molecular fluorine or xenon difluoride in the presence of BF(3).OEt(2). Among compounds 3a-d, the 4-O-thiocarbamoylmethyl derivative 3c showed the most potent inhibitory activity against sialidase of human parainfluenza virus type 1. [structure: see text]. PMID- 15113677 TI - Improved anomeric selectivity for the aroylation of sugars. AB - By manipulating the solvent and using bulky TMEDA as a base, good yields and improved anomeric selectivities were obtained for the aroylation of D-glucose over similar esterifications using pyridine. The reaction has been extended to mannose and the beta-anomer of pergalloylated mannose was predominantly obtained in one step by direct aroylation of the parent sugar. PMID- 15113678 TI - Palladium-catalyzed oxidation of benzylated aldose hemiacetals to lactones. AB - Benzyl protected sugar hemiacetals are oxidized to the corresponding lactones in excellent yields using bromobenzene, K(2)CO(3) and the Pd(OAc)(2)/PPh(3) catalytic system. PMID- 15113679 TI - Preparation of polyhydroxyalkyl- and C-glycosylfuran derivatives from free sugars catalyzed by cerium(III) chloride in aqueous solution: an improvement of the Garcia Gonzalez reaction. AB - Unprotected aldose sugars react smoothly with 1,3-diones or beta-ketoesters in the presence of CeCl(3).7H(2)O in aqueous solution to produce polyhydroxylalkyl- and C-glycosylfuran derivatives in excellent yield. Operationally simple, mild neutral reaction conditions in aqueous solution is the key feature of this methodology. PMID- 15113680 TI - alpha-Mannosidase-catalyzed synthesis of a (1-->2)-alpha-D-rhamnodisaccharide derivative. AB - Enzymatic transglycosylation using p-nitrophenyl alpha-D-rhamnopyranoside as the glycosyl donor and 6equiv of ethyl 1-thio-alpha-D-rhamnopyranoside as the glycosyl acceptor yielded a D-rhamnooligosaccharide derivative. The reaction was catalyzed by jack bean alpha-mannosidase in a 1:1 (v/v) mixture of 0.1 M sodium citrate buffer (pH4.5)-MeCN at 25 degrees C. The enzyme exhibited high catalytic activity for the reaction, to afford in 32.1% isolated yield (based on donor substrate) ethyl alpha-D-rhamnopyranosyl-(1-->2)-1-thio-alpha-D-rhamnopyranoside, which is a derivative of the common oligosaccharide unit of the antigenic lipopolysaccharides from Pseudomonas. PMID- 15113681 TI - Structure of the O-polysaccharide of Proteus mirabilis CCUG 10701 (OB) classified into a new Proteus serogroup, O74. AB - An acidic O-polysaccharide was isolated by mild acid degradation of the lipopolysaccharide of Proteus mirabilis CCUG 10701 (OB) and studied by chemical analyses and (1)H and (13)C NMR spectroscopy. The following structure of the tetrasaccharide repeating unit of the polysaccharide was established: --> 3)-beta D-GlcpNAc6Ac-(1 --> 2)-beta-D-GalpA4Ac-(1--> 3)-alpha-D-GalpNAc-(1 --> 4)-alpha-D GalpA-(1 -->, where the degree of O-acetylation at position 6 of GlcNAc is approximately 50% and at position 4 of beta-GalA approximately 60%. Based on the unique structure of the O-polysaccharide and serological data, it is proposed to classify P. mirabilis CCUG 10701 (OB) into a new Proteus serogroup, O74. PMID- 15113682 TI - Structural characterization of the antigenic O-polysaccharide in the lipopolysaccharide produced by Actinobacillus pleuropneumoniae serotype 14. AB - The antigenic O-polysaccharide of the lipopolysaccharide produced by Actinobacillus pleuropneumoniae serotype 14 was shown by chemical analysis and 1D and 2D nuclear magnetic resonance methods to be a high-molecular-mass polymer of a repeating disaccharide unit composed of a chain of (1-->5)-linked beta-D galactofuranose (beta-D-Galf) residues substituted at their O-2 positions by alpha-D-galactopyranose residues (D-Galp) (1:1): [Formula: see text]. PMID- 15113683 TI - Transglycosylation reaction of Mucor hiemalis endo-beta-N-acetylglucosaminidase using sugar derivatives modified at C-1 or C-2 as oligosaccharide acceptors. AB - We investigated the transglycosylation reaction of the recombinant endo-beta-N acetylglucosaminidase from Mucor hiemalis (Endo-M) expressed in Candida boidinii using such sugar derivatives as N-acylated d-glucosamines, C-glucosyl derivatives, and a 2-O-glycosylated disaccharide as acceptors. We found that a variety of sugar derivatives modified at C-1 or C-2 could be used as acceptors for transglycosylation by Endo-M to create novel oligosaccharides. PMID- 15113684 TI - Gene expression analysis by real-time reverse transcription polymerase chain reaction: influence of tissue handling. AB - Factors such as warm ischemia and time at room temperature before tissue treatment may influence the results of mRNA expression analyses on tissue specimens obtained during surgery. We evaluated the effect of these factors on RNA integrity and mRNA expression levels by incubating freshly obtained mouse liver tissue at 25 or 37 degrees C for periods of 0-4 h. Changes in the mRNA expression levels of seven genes, Tbp, Eef1a, Fos, Junb, Myc, Vegf, and Glut2, were determined by real-time reverse transcription-polymerase chain reaction. Incubation at 25 degrees C for up to 4 h only slightly altered (by a factor of less than 2) levels of mRNA for Tbp, Eef1a, Junb, Myc, Vegf, and Glut2. This result is consistent with limited RNA degradation at this temperature. Incubation at 37 degrees C strongly affected the levels of these mRNAs. Four hours of incubation at this temperature resulted in extensive RNA degradation, with mRNA levels falling to 1/10th those before incubation. When relative quantification was performed, i.e., quantification of the target gene transcripts in comparison to an endogenous housekeeping transcript (Tbp or Eef1a), the changes in mRNA levels were reduced to less than 2.5-fold. Fos behaved very differently from the other genes tested on incubation, with Fos mRNA levels increasing considerably following incubation at either 25 or 37 degrees C. Our data suggest that, with the exception of certain genes induced by tissue injury, relative quantification of mRNA, even on degraded RNA samples, can provide a reliable estimate of in vivo mRNA levels. PMID- 15113685 TI - Interference of calmidazolium with measurement of mitochondrial membrane potential using the tetraphenylphosphonium electrode or the fluorescent probe rhodamine 123. AB - Calmidazolium (CMZ) is a positively charged, hydrophobic compound used as a calmodulin antagonist. It may cause unspecific effects in mitochondria, e.g., a decrease in membrane potential (deltapsi), swelling, and uncoupling. Several groups have advised against use of CMZ in studying signal transduction in mitochondria. We report here that it interferes with measurement of deltapsi in rat liver mitochondria (RLM) when using the tetraphenyl phosphonium (TPP+) electrode. We also found that CMZ reduces the signal, indicating an apparent drop in deltapsi. CMZ itself gave a signal with the TPP+ electrode in the absence of RLM. At high concentrations, > 10 microM, it also reduced the fluorescence quenching of the probe rhodamine 123. This may be due to an interference with mitochondrial uptake and binding of this positively charged probe or to an uncoupling effect. It is concluded that CMZ and similar positively charged calmodulin antagonists such as trifluoperazine may be used in mitochondria if these interferences are controlled and calibration is carried out under the experimental conditions used. PMID- 15113686 TI - Microbead-based electrochemical immunoassay with interdigitated array electrodes. AB - The objective of this study was to develop a sensitive and miniaturized immunoassay by coupling a microbead-based immunoassay with an interdigitated array (IDA) electrode. An IDA electrode amplifies the signal by recycling an electrochemically redox-reversible molecule. The microfabricated platinum electrodes had 25 pairs of electrodes with 1.6-microm gaps and 2.4-microm widths. An enzyme-labeled sandwich immunoassay on paramagnetic microbeads with mouse IgG as the analyte and beta-galactosidase as the enzyme label was used as the model system. beta-Galactosidase converted p-aminophenyl beta-D-galactopyranoside to p aminophenol (PAP). This enzyme reaction was measured continuously by positioning the microbeads near the electrode surface with a magnet. Electrochemical recycling occurred with PAP oxidation to p-quinone imine (PQI) at +290 mV followed by PQI reduction to PAP at -300 mV vs Ag/AgCl. Dual-electrode detection amplified the signal fourfold compared to single-electrode detection, and the recycling efficiency reached 87%. A calibration curve of PAP concentration vs anodic current was linear between 10(-4) and 10(-6)M. A signal from 1000 beads in a 20-microL drop was detectable and the immunoassay was complete within 10 min with a detection limit of 3.5x10(-15)mol mouse IgG. PMID- 15113687 TI - Stability parameters for one-step mechanism of irreversible protein denaturation: a method based on nonlinear regression of calorimetric peaks with nonzero deltaCp. AB - Thermal transitions of many proteins have been found to be calorimetrically irreversible and scan-rate dependent. Calorimetric determinations of stability parameters of proteins which unfold irreversibly according to a first-order kinetic scheme have been reported. These methods require the approximation that the increase in heat capacity upon denaturation deltaCp is zero. A method to obtain thermodynamic parameters and activation energy for the two-state irreversible process N --> D from nonlinear fitting to calorimetric traces is proposed here. It is based on a molar excess heat capacity function which considers irreversibility and a nonzero constant deltaCp. This function has four parameters: (1) temperature at which the calorimetric profile reaches its maximal value (Tm), (2) calorimetric enthalpy at Tm (deltaHm), (3) deltaCp, and (4) activation energy (E). The thermal irreversible denaturation of subtilisin BPN' from Bacillus amyloliquefaciens was studied by differential scanning calorimetry at pH 7.5 to test our model. Transitions were found to be strongly scanning-rate dependent with a mean deltaCp value of 5.7 kcal K(-1)mol(-1), in agreement with values estimated by accessible surface area and significantly higher than a previously reported value. PMID- 15113688 TI - Discovery of novel inhibitors of Bcl-xL using multiple high-throughput screening platforms. AB - Bcl-xL is a member of the Bcl-2 family of proteins that are implicated to play a vital role in several diseases including cancer. Bcl-xL suppresses apoptosis; thus the inhibition of Bcl-xL function could restore the apoptotic process. To identify antagonists of Bcl-xL function, two ultra-high-throughput screens were implemented. An activity assay utilized fluorescence polarization, based on the binding of fluorescein-labeled peptide [the BH3 domain of BAD protein (F-Bad 6)] to Bcl-xL. A 384-well plate assay with mixtures of 10 drug compounds per well, combined with a fast plate reader, resulted in a throughput of 46,080 data points/day. Utilizing this screening format, 370,400 compounds were screened in duplicate and 425 inhibitors with an IC(50) below 100 microM were identified. The second assay format, affinity selection/mass spectrometry (ASMS), used ultrafiltration to separate Bcl-xL binders from nonbinders in mixtures of 2400 compounds. The bound species were subsequently separated from the protein and analyzed by flow injection electrospray mass spectrometry. Utilizing the ASMS format, 263,382 compounds were screened in duplicate and 29 binders with affinities below 100 microM were identified. Two novel classes of Bcl-xL inhibitors were identified by both methods and confirmed to bind (13)C-labeled Bcl-xL using heteronuclear magnetic resonance spectroscopy. PMID- 15113689 TI - Quantification of polyphosphate: different sensitivities to short-chain polyphosphate using enzymatic and colorimetric methods as revealed by ion chromatography. AB - Polyphosphate is ubiquitous and has a variety of biochemical functions. Among polyphosphate quantification methods, an enzymatic assay using Escherichia coli polyphosphate kinase (PPK), in which polyphosphate is converted to adenosine 5' triphosphate and quantified by luciferase assay, is the most specific and most sensitive. However, chain-length specificity of the assay has not been analyzed in detail so far. Ion chromatography equipped with an on-line hydroxide eluent generator enabled us to analyze polyphosphate up to 50 inorganic phosphate (P(i)) residues, and we employed this method to investigate the chain-length specificity of PPK in this study. Several fractions of short-chain polyphosphate were prepared by electrophoresis, and the chain-length distribution was analyzed before and after 1-6 h PPK reaction by ion chromatography. Polyphosphates longer than 23 P(i) residues were processed by PPK completely after 1 h incubation, but complete processing of those between 11 and 22 P(i) residues required 6h incubation. Limited processing of polyphosphates of 10 P(i) residues or shorter were observed even after 6h incubation. Metachromasy of Toluidine blue O, an alternative method for polyphosphate quantification, showed broader chain-length specificity although it was not as sensitive as the enzymatic assay. Combination of these two methods would be practically applicable to analysis of polyphosphate dynamics in living organisms. PMID- 15113690 TI - Sesquiterpene lactones inhibit luciferase but not beta-galactosidase activity in vitro and ex vivo. AB - Reporter enzymes such as firefly luciferase or beta-galactosidase of Escherichia coli are frequently used to study transcriptional activity of genes and to investigate the effects of novel compounds on gene or transcription factor activity. It is generally assumed that the activity of these enzymes is unaffected by the treatment conditions. Therefore, this factor is not considered when interpreting the data obtained. Biologically active compounds such as sesquiterpene lactones (SLs) have also been tested in reporter gene assays for their influence on gene expression. Here we show in in vitro and ex vivo experiments that SLs inhibit firefly luciferase activity probably by direct targeting of the enzyme while beta-galactosidase remains almost completely unaffected. The loss of luciferase activity after SL treatment could be an effect of their sulfhydryl-modifying potency and the subsequent alteration of the enzyme's tertiary structure. These results demonstrate that the effect of the test substance on the reporter enzyme used should be taken into consideration when the transcriptional effect of novel compounds is investigated. PMID- 15113691 TI - Characterization of major histocompatibility complex-associated peptides from a small volume of whole blood. AB - Class I major histocompatibility complex (MHC) presents intracellular-derived peptides on the majority of cells within the human body. Intracellular proteins are degraded into peptides of 8-11 amino acids, allowing them to fit into the groove of an empty MHC class I molecule. Detection of MHC-associated peptides can be challenging with the major difficulty being the ability to obtain peptides in adequate concentration. Published protocols require a large sample size that is unrealistic for a clinically available sample. Based on calculations, it should be possible to characterize MHC-associated peptides from cells obtained from 30 ml of whole blood. A citric acid wash of whole platelets was implemented to release the peptides with sample cleanup by reversed-phase high-performance liquid chromatography on a peptide trap. Peptides were analyzed by liquid chromatography tandem mass spectrometry. Four peptides were identified from an individual's platelets. The binding motifs of the peptides were consistent with the published MHC binding motif of the individual. Since red blood cells do not express MHC, they were used as a negative control. Using citric acid wash of whole cells and a peptide trap, the more abundant MHC-associated peptides can be identified. This report demonstrates the identification of peptides from a sample volume compatible with reasonable clinical availability. PMID- 15113692 TI - Coated microwell plate-based affinity purification of antigens. AB - Antibody-based affinity capture of antigens is widely used in the isolation of antigens from complex mixtures. Antibody and the corresponding antigen are allowed to interact with each other to form immunocomplexes which are then typically captured by protein A or protein G immobilized on beaded support. Antigen capture performed using this method generally requires multiple centrifugation steps and careful pipetting to avoid loss of the bead-bound complexes. This traditional procedure is tedious and not easily reproducible, especially when working with multiple samples. To address these issues we have demonstrated that antigens can be captured with protein A/G, protein G, and high binding-capacity streptavidin 96-well strip-coated plates. The coated plate method of antigen purification is reproducible, within the same experiment and between experiments, due to the uniform binding capacity of the plates and wells. Here we report the use of coated microwell plates for antigen capture and for protein-protein interaction studies with the well-characterized BIR2-SMAC, transferrin receptor/ transferrin, and other systems. PMID- 15113693 TI - Characterization of Mca-Lys-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2, a fluorogenic substrate with increased specificity constants for collagenases and tumor necrosis factor converting enzyme. AB - Matrix metalloproteinases (MMPs) and the related tumor necrosis factor converting enzyme (TACE) are involved in tissue remodeling, cell migration, and processing of signaling molecules, such as cytokines and adhesion molecules. Fluorescence quenched peptide substrates have been widely used to quantitate the actual enzymatic activity of MMPs. However, the various MMPs have very different specific activities toward these substrates. This restricts their value for the determination of composite proteolytic activity of mixtures of metalloproteinases in biological fluids. The N-terminal elongation of the most widely used MMP substrate (FS-1) with a Lys to the sequence Mca-Lys-Pro-Leu-Gly-Leu-Dpa-Ala-Arg NH(2) (FS-6) yields a fluorogenic peptide with improved substrate properties. As compared to FS-1, the specificity constant (kcat/Km) of FS-6 for collagenases (MMP-1, MMP-8, MMP-13) and MT1-MMP (MMP-14) is increased two- to ninefold and threefold, respectively, while those for gelatinases and matrilysin remain equally high. Using high-performance liquid chromatography-fluorescence detection, MMP activity can be quantitated in the picomolar range. FS-6 shows up to twofold higher specificity constants (kcat/Km of 0.8x10(6)M(-1)s(-1)) for TACE, as compared to standard substrates Mca-PLAQAV-Dpa-RSSSAR-NH(2) and Dabcyl LAQAVRSSSAR-EDANS. FS-6 is fully water soluble and thus allows measurement of metalloproteinase activity in tissue culture conditions, e.g., on the surface of viable cells in situ. PMID- 15113694 TI - Use of the fluorescent dye 10-N-nonyl acridine orange in quantitative and location assays of cardiolipin: a study on different experimental models. AB - The fluorescent dye 10-N-nonyl acridine orange (NAO) is extensively used for location and quantitative assays of cardiolipin in living cells on the assumption of its high specificity for cardiolipin; however, the limits and the mechanism of this specificity are not clear. Moreover, whether factors such as the membrane potential in mitochondria may limit the consistency of the results obtained by this method is open to discussion. The aim of this research was to investigate the effects of some experimental factors on the selective fluorescence of NAO in the presence of cardiolipin in artificial and natural membranes (mitochondria). The results show that the fluorescence of NAO, due to interaction with cardiolipin, is significantly modified by factors that control the spatial arrangement of cardiolipin molecules within the space of the membrane under investigation. Moreover, the present observations suggest that the specific effect of cardiolipin is to facilitate the dimerization of this fluorescent dye, thus confirming that reliable measurements of cardiolipin concentration can be obtained only when the NAO/cardiolipin molar ratio is equal to 2. The finding is also reported that in isolated respiring mitochondria the interaction of NAO with cardiolipin is somewhat related to the respiratory state of mitochondria. PMID- 15113695 TI - Electrooptical analysis of the Escherichia coli-phage interaction. AB - This article describes electrooptical (EO) characterization of biospecific binding between the bacterium Escherichia coli XL-1 and the phage M13K07. The electrooptical analyzer (ELUS EO), which has been developed at the State Research Center for Applied Microbiology, Obolensk, Russia, was used as the basic instrument for EO measurements. The operating principle of the analyzer is based on the polarizability of microorganisms, which depends strongly on their composition, morphology, and phenotype. The principle of analysis of the interaction of E. coli with the phage M13K07 is based on registration of changes of optical parameters of bacterial suspensions. The phage-cell interaction includes the following stages: phage adsorption on the cell surface, entry of viral DNA into the bacterial cell, amplification of phage within infected host, and phage ejection from the cell. In this work, we used M13K07, a filamentous phage of the family Inoviridae. Preliminary study had shown that combination of the EO approach with a phage as a recognition element has an excellent potential for mediator-less detection of phage-bacteria complex formation. The interaction of E. coli with phage M13K07 induces a strong and specific EO signal as a result of substantial changes of the EO properties of the E. coli XL-1 suspension infected by the phage M13K07. The signal was specific in the presence of foreign microflora (E. coli K-12 and Azospirillum brasilense Sp7). Integration of the EO approach with a phage has the following advantages: (1) bacteria from biological samples need not be purified, (2) the infection of phage to bacteria is specific, (3) exogenous substrates and mediators are not required for detection, and (4) it is suitable for any phage-bacterium system when bacteria-specific phages are available. PMID- 15113696 TI - Europium-labeled melanin-concentrating hormone analogues: ligands for measuring binding to melanin-concentrating hormone receptors 1 and 2. AB - We investigated the use of Eu3+ chelate-labeled analogues of melanin concentrating hormone (MCH) as ligands for both human MCH receptors (MCHR1 and MCHR2). The analogues employed were Ala17 MCH, S36057 (Y-ADO-RC*MLGRVFRPC*W, where ADO=8-amino-3,6-dioxyoctanoyl and *=disulfide bond), and R2P (RC*MLGRVFRPC*Y-NH2). The peptides were readily labeled on the alpha-amino residue with the Eu3+ chelate of N1-(p-isothiocyanatobenzyl)-diethylenetriamine N1,N2,N3,N3-tetraacetic acid and then purified by reverse-phase fast-performance liquid chromatography at neutral pH to maintain Eu3+ chelation. Both labeled Ala17 MCH and S36057 had high affinity for MCHR1 ( Kd = 0.37 and 0.059nM, respectively) while Eu3+ -labeled S36057 and R2P had high affinity for MCHR2 ( Kd = 0.16 and 0.10nM, respectively). Labeled Ala17 MCH had little demonstrable binding affinity for MCHR2. Eu3+ -labeled S36057 and R2P were full agonists at MCHR1 when assessed by measurement of agonist-stimulated GTPgamma(35)S binding. Competition binding experiments with both MCHR isoforms, a series of previously characterized alanine scan MCH analogues, and a recently identified nonpeptide MCHR1-selective antagonist T-226296 confirmed the expected receptor selectivity. These studies further extend the utility of Eu3+ chelate time-resolved fluorescence for the development of high-sensitivity, nonradioactive receptor binding assays and demonstrate the need to select the optimal ligand for labeling. PMID- 15113697 TI - General assay for sugar nucleotidyltransferases using electrospray ionization mass spectrometry. AB - An electrospray ionization mass spectrometry-based assay has been developed to study the class of enzymes called sugar nucleotidyltransferases that couple sugar 1-phosphates and nucleotide triphosphates to form Leloir pathway glycosyl donors. The recombinant Escherichia coli and the commercially available yeast uridine diphosphoglucose pyrophosphorylases were used as model systems. This technique allows the simultaneous and direct detection of the substrates and products without separation and, as described, is as sensitive as traditional coupled techniques. More importantly, the assay is capable of easily measuring kinetic values and inhibition constants for a range of natural and nonnatural substrates. This new assay was used to show for the first time that the reaction of the commercially available yeast uridine-diphosphoglucose pyrophosphorylase preparation is competitively inhibited by adenosine 5'-triphosphate (ATP), an observation that indicates a single active site that accepts both uridine 5' triphosphate and ATP substrates. PMID- 15113698 TI - Isolation and identification of two isomeric forms of malonyl-coenzyme A in commercial malonyl-coenzyme A. AB - Two isomers of malonyl-coenzyme A (malonyl-CoA) were detected in a commercial preparation of malonyl-CoA. These compounds were separated by preparative high performance liquid chromatography (HPLC) and characterized by HPLC/ultraviolet (UV)/mass spectrometry. Both compounds had a UV absorbance maximum at 259-260 nm. Both compounds underwent negative electrospray ionization to produce a [M-H]( )quasi-molecular ion at m/z 852 and both compounds underwent collision-induced dissociation to produce a characteristic fragment at m/z 808, all consistent with the structure of malonyl-CoA. Nuclear magnetic resonance spectrometry showed that the two chromatographically distinguishable malonyl-CoAs are structural isomers: the major component is the naturally occurring malonyl-CoA and the contaminant is 3'-dephospho- 2'-phospho-coenzyme A. PMID- 15113699 TI - Fluorescent nanoparticles as labels for immunometric assay of C-reactive protein using two-photon excitation assay technology. AB - We describe the use of fluorophore-doped nanoparticles as reporters in a recently developed ArcDia TPX bioaffinity assay technique. The ArcDia TPX technique is based on the use of polymer microspheres as solid-phase reaction carrier, fluorescent bioaffinity reagents, and detection of two-photon excited fluorescence. This new assay technique enables multiplexed, separation-free bioaffinity assays from microvolumes with high sensitivity. As a model analyte we chose C-reactive protein (CRP). The assay of CRP was optimized for assessment of CRP baseline levels using a nanoparticulate fluorescent reporter, 75 nm in diameter, and the assay performance was compared to that of CRP assay based on a molecular reporter of the same fluorophore core. The results show that using fluorescent nanoparticles as the reporter provides two orders of magnitude better sensitivity (87 fM) than using the molecular label, while no difference between precision profiles of the different assay types was found. The new assay method was applied for assessment of baseline levels of CRP in sera of apparently healthy individuals. PMID- 15113700 TI - Factors that affect the efficiency of cell transfection by immunoporation. AB - Immunoporation is a recently discovered method that is able to transfect various human cell lines efficiently by targeting the cell surface antigens with antibody coated beads. For this particular study, HL60, a cell line difficult to transfect by other methods, was used as a model to define the various parameters of the cell membrane that determine the efficiency of this method. The level of antigen expression on the cell surface was the first parameter to be analyzed and experiments indicated that there is a close correlation between the level of expression of surface antigens and the efficiency of immunoporation. The mixing speed, the bead to cell ratio, and the mixing time were all found to affect the ability of the antigen-coated beads to pull holes in the cells and it was found that for HL60 cells the optimum mixing speed was 40 rpm and the bead to cell ratio was 20:1 using a mixing time of 6 h. PMID- 15113701 TI - A continuous spectrophotometric assay for Pseudomonas aeruginosa LasA protease (staphylolysin) using a two-stage enzymatic reaction. AB - Pseudomonas aeruginosa LasA protease is a secreted metalloendopeptidase that can lyse Staphylococcus aureus cells by cleaving the pentaglycine bridges of their peptidoglycan. It can also degrade elastin and stimulate shedding of cell-surface proteoglycans, activities implicated in pathogenesis of P. aeruginosa infections. The activity of LasA protease can be assayed spectrophotometrically by following the reduction in turbidity of S. aureus cell suspensions. This assay, however, does not permit kinetic studies and its reproducibility is poor. Here we describe a two-stage enzymatic reaction for the continuous measurement of LasA protease activity using a defined substrate, succinyl-Gly-Gly-Phe-4-nitroanilide, supplemented with Streptomyces griseus aminopeptidase. Cleavage of the Gly-Phe bond by LasA protease is followed by hydrolysis of the product Phe-4-nitroanilide by the aminopeptidase and the rate of release of the chromophore (4-nitroaniline) is measured spectrophotometrically using a 96-well microplate reader. Activity of nanogram amounts of LasA protease could be determined within a few minutes. Furthermore, this assay permitted the determination of Km and kcat values for LasA protease, which were 0.46 mM and 11.8s(-1), respectively. Pseudomonas elastase was also active in the assay. However, it was less effective than LasA protease and its activity was inhibited by phosphoramidon. The assay is highly sensitive and reproducible, providing a convenient tool for further studies of LasA protease function(s) and mechanism of action. PMID- 15113702 TI - Probing the mechanism of drug/lipid membrane interactions using Biacore. AB - Assay conditions were established to screen a panel of drugs for binding to liposome surfaces using a surface plasmon resonance (SPR) biosensor. Drugs were found to bind negligibly or reversibly or were retained on the liposome surface. Cationic amphiphilic drugs fell into the last class and correlated with drugs that induce phospholipidosis in vivo. To a first approximation, a single-site model yielded apparent binding affinities that adequately described a drug's dose dependent binding to liposome surfaces. Affinities ranged at least 1000-fold within the drug panel. A liposome's drug-binding capacity and affinity depended on both the lipid headgroup and the drug's structure. Although a drug's charge state generally dominated whether or not it remained bound to the liposome, subtle structural differences between members of certain drug families led to them having widely differing binding affinities. A comparison between the dissociation of drugs from liposome surfaces by Biacore and the lipid retention measurements determined by a parallel artificial membrane permeability assay was drawn. The results from this study demonstrate the potential of using SPR-based assays to characterize drug/liposome-binding interactions. PMID- 15113703 TI - Prenatal programming of adult thyroid function by alcohol and thyroid hormones. AB - Increasing evidence associates environmental challenges early in life with permanent alterations of physiological functions in adulthood. These changes in fetal environment can trigger physiological adaptations by the fetus, called fetal programming, which may be beneficial before birth but permanently influence the physiology of the organism. In this study, we investigated the potential connection between alcohol-induced decreased maternal thyroid function and the hypothalamic-pituitary-thyroid (HPT) function of adult rat offspring. Plasma 3,5,3'-triiodothyronine (T(3)), thyroxine (T(4)), and thyroid-stimulating hormone (TSH) levels were decreased in alcohol-consuming (E) dams on gestational day 21 compared with ad libitum- (C) and pair-fed (PF) controls. No significant differences were found in HPT function in young offspring (3 wk of age) between diet groups. However, adult fetal alcohol-exposed (FAE) offspring had significantly decreased levels of T(3) along with elevated TSH compared with control offspring. T(4) administration to the mother did not normalize the hypothyroid state of the adult FAE offspring. Interestingly, administration of T(4) to control pregnant dams decreased plasma T(3) of the adult female offspring only, whereas T(4) together with maternal alcohol consumption or pair-feeding led to decreased TSH and T(4) in the adult female offspring. Our results suggest that ethanol consumption and T(4) administration alter maternal HPT function, leading to prenatally programmed permanent alterations in the thyroid function of the adult offspring. PMID- 15113704 TI - Functional characterization of an insulin-responsive glucose transporter (GLUT4) from fish adipose tissue. AB - Glucose transport across the plasma membrane is mediated by a family of glucose transporter proteins (GLUTs), several of which have been identified in mammalian, avian, and, more recently, in fish species. Here, we report on the cloning of a salmon GLUT from adipose tissue with a high sequence homology to mammalian GLUT4 that has been named okGLUT4. Kinetic analysis of glucose transport following expression in Xenopus laevis oocytes demonstrated a 7.6 +/- 1.4 mM K(m) for 2 deoxyglucose (2-DG) transport measured under zero-trans conditions and 14.4 +/- 1.5 mM by equilibrium exchange of 3-O-methylglucose. Transport of 2-DG by okGLUT4 injected oocytes was stereospecific and was competed by D-glucose, D-mannose, and, to a lesser extent, D-galactose and D-fructose. In addition, 2-DG uptake was inhibited by cytochalasin B and ethylidene glucose. Moreover, insulin stimulated glucose uptake in Xenopus oocytes expressing okGLUT4 and in isolated trout adipocytes, which contain the native form of okGLUT4. Despite differences in protein motifs important for insulin-stimulated translocation of mammalian GLUT4, okGLUT4 was able to translocate to the plasma membrane from intracellular localization sites in response to insulin when expressed in 3T3-L1 adipocytes. These data demonstrate that okGLUT4 is a structural and functional fish homolog of mammalian GLUT4 but with a lower affinity for glucose, which could in part explain the lower ability of fish to clear a glucose load. PMID- 15113705 TI - Longitudinal changes in energy expenditure and body composition in obese women with normal and impaired glucose tolerance. AB - Our primary objective was to evaluate changes in energy expenditure and body composition in women with normal glucose tolerance (NGT) and gestational diabetes mellitus (GDM). A secondary objective was to examine the relationship between maternal leptin and nutrient metabolism. Fifteen obese women, eight with NGT and seven with GDM, were evaluated before conception (P), at 12-14 wk (E), and at 34 36 wk (L). Energy expenditure and glucose and fat metabolism were measured using indirect calorimetry. Basal hepatic glucose production was measured using [6,6 2H2]glucose and insulin sensitivity by euglycemic clamp. There was a significant increase (6.6 kg, P = 0.0001) in fat mass from P to L. There was a 30% (P = 0.0001) increase in basal O2 consumption (VO2, ml/min). There were no significant changes in carbohydrate oxidation during fasting or storage from P to L. There was, however, a significant (P = 0.0001) 150% increase in basal fat oxidation (mg/min) from P to L. Under hyperinsulinemic conditions, there were similar 25% increases in VO2 (P = 0.0001) from P to L in both groups. Because of the significant increases in insulin resistance from P to L, there was a significant (P = 0.0001) decrease in carbohydrate oxidation and storage. There was a net change from lipogenesis to lipolysis, i.e., fat oxidation (30-40 mg/min, P = 0.0001) from P to L. Serum leptin concentrations had a significant positive correlation with fat oxidation at E (r = 0.76, P = 0.005) and L (r = 0.72, P = 0.009). Pregnancy in obese women is associated with significant increases in fat mass and basal metabolic rate and an increased reliance on lipids both in the basal state and during the clamp. These modifications are similar in women with NGT and GDM. The increased reliance on fat metabolism is accompanied by a concomitant decrease in carbohydrate metabolism during hyperinsulinemia. The increase in fat oxidation may be related to increased maternal serum leptin. PMID- 15113706 TI - Too much sugar, too much carbohydrate, or just too much? PMID- 15113707 TI - A healthy lifestyle lowers homocysteine, but should we care? PMID- 15113708 TI - Nutrient selection and the genetics of complex phenotypes. PMID- 15113709 TI - Assessment of dietary vitamin D requirements during pregnancy and lactation. AB - Concerns about vitamin D have resurfaced in medical and scientific literature because the prevalence of vitamin D deficiency in the United States, particularly among darkly pigmented persons, has increased. The primary goals of this review were to discuss past and current literature and to reassess the dietary reference intake for vitamin D in adults, with particular focus on women during pregnancy and lactation. The appropriate dose of vitamin D during pregnancy and lactation is unknown, although it appears to be greater than the current dietary reference intake of 200-400 IU/d (5-10 microg/d). Doses of < or =10 000 IU vitamin D/d (250 microg/d) for up to 5 mo do not elevate circulating 25-hydroxyvitamin D to concentrations > 90 ng/mL, whereas doses < 1000 IU/d appear, in many cases, to be inadequate for maintaining normal circulating 25-hydroxyvitamin D concentrations of between 15 and 80 ng/mL. Vitamin D plays no etiologic role in cardiac valvular disease, such as that observed in Williams syndrome, and, as such, animal models involving vitamin D intoxication that show an effect on cardiac disease are flawed and offer no insight into normal human physiology. Higher doses of vitamin D are necessary for a large segment of Americans to achieve concentrations equivalent to those in persons who live and work in sun-rich environments. Further studies are necessary to determine optimal vitamin D intakes for pregnant and lactating women as a function of latitude and race. PMID- 15113710 TI - Polyphenols: food sources and bioavailability. AB - Polyphenols are abundant micronutrients in our diet, and evidence for their role in the prevention of degenerative diseases such as cancer and cardiovascular diseases is emerging. The health effects of polyphenols depend on the amount consumed and on their bioavailability. In this article, the nature and contents of the various polyphenols present in food sources and the influence of agricultural practices and industrial processes are reviewed. Estimates of dietary intakes are given for each class of polyphenols. The bioavailability of polyphenols is also reviewed, with particular focus on intestinal absorption and the influence of chemical structure (eg, glycosylation, esterification, and polymerization), food matrix, and excretion back into the intestinal lumen. Information on the role of microflora in the catabolism of polyphenols and the production of some active metabolites is presented. Mechanisms of intestinal and hepatic conjugation (methylation, glucuronidation, sulfation), plasma transport, and elimination in bile and urine are also described. Pharmacokinetic data for the various polyphenols are compared. Studies on the identification of circulating metabolites, cellular uptake, intracellular metabolism with possible deconjugation, biological properties of the conjugated metabolites, and specific accumulation in some target tissues are discussed. Finally, bioavailability appears to differ greatly between the various polyphenols, and the most abundant polyphenols in our diet are not necessarily those that have the best bioavailability profile. A thorough knowledge of the bioavailability of the hundreds of dietary polyphenols will help us to identify those that are most likely to exert protective health effects. PMID- 15113711 TI - Skeletal muscle lipid concentration quantified by magnetic resonance imaging. AB - BACKGROUND: Skeletal muscle lipid is associated with obesity and type 2 diabetes and may be altered by diet, physical activity, and weight loss. OBJECTIVE: We explored the utility of magnetic resonance imaging (MRI) for quantifying the lipid concentration of muscle tissue in vivo. DESIGN: Fat-selective MR images of the lower leg were taken in 8 normal-weight [body mass index (in kg/m(2)) < or = 24.9] and 8 obese (body mass index > 29.9) subjects to obtain spatial maps of lipid signal intensity within muscle tissue. Fast-spiral-sequence (echo time = 5.6-13.8 ms, repetition time = 1 s, 8 interleaves) MRI scans were conducted at 3.0 T by using an extremity transmit-receive coil. Lipid concentrations within muscle were determined from manually drawn regions of interest in the tibialis anterior (TA), soleus, and medial head of the gastrocnemius (MHG) muscle groups. RESULTS: There was extremely good agreement (mean R(2) = 0.985) between the fat signal intensity and the actual lipid concentration of standards containing 2.5, 5.0, and 10.0 g lipid/dL, which were placed on the subject's leg during each scan. The lipid content of both the soleus (2.99 +/- 0.37 g/dL) and the MHG (3.80 +/- 0.68 g/dL) was higher (P < 0.05) than that of the TA (1.83 +/- 0.28 g/dL). Lipid content was more than two-fold higher (P < 0.05) in the MHG of obese subjects (5.48 +/- 1.18 g/dL) than in the MHG of normal-weight subjects (2.54 +/- 0.47 g/dL), but did not differ significantly in the TA or soleus. CONCLUSIONS: MRI can be used to quantify lipid within human muscle tissue. MRI can also be used to detect differences in muscle lipid content among various muscle groups and between normal-weight and obese subjects. PMID- 15113712 TI - Dietary fat consumption and primary open-angle glaucoma. AB - BACKGROUND: Prostaglandin F(2alpha) analogues are effective intraocular-pressure lowering drugs. Dietary fatty acids affect endogenous prostaglandin F(2alpha) concentrations and may thus influence intraocular pressure. OBJECTIVE: We prospectively examined dietary fat consumption in relation to primary open-angle glaucoma (POAG). DESIGN: Women (n = 76 199 in the Nurses' Health Study) and men (n = 40 306 in the Health Professionals Follow-Up Study) free of POAG in 1980 and 1986, respectively, were followed until 1996 if they were > or =40 y old and reported receiving eye exams during follow-up. Potential confounders were assessed on biennial questionnaires, and energy-adjusted cumulative averaged fat intakes were measured by using validated food-frequency questionnaires. We analyzed 474 self-reported POAG cases confirmed by medical chart review. Cohort specific multivariate rate ratios (RRs) were obtained by using proportional hazards models and were then pooled. RESULTS: Major fats and fat subtypes were not independently associated with POAG risk. Pooled multivariate RRs (95% CI) for POAG comparing the highest with the lowest quintile of fat intake were as follows: 0.90 (0.67, 1.21) for total fat, 1.03 (0.77, 1.38) for saturated fat, 0.76 (0.56, 1.03) for monounsaturated fat, and 0.87 (0.66, 1.16) for polyunsaturated fat, none of which were statistically significant. We found a suggestive positive association between a higher ratio of n-3 to n-6 polyunsaturated fat and risk of POAG [RR = 1.49 (1.11, 2.01); P for trend = 0.10], which was stronger for high-tension POAG [RR = 1.68 (1.18, 2.39); P for trend = 0.009]. CONCLUSION: A high ratio of n-3 to n-6 polyunsaturated fat appears to increase the risk of POAG, particularly high-tension POAG. Further studies are needed. PMID- 15113713 TI - Differential eicosapentaenoic acid elevations and altered cardiovascular disease risk factor responses after supplementation with docosahexaenoic acid in postmenopausal women receiving and not receiving hormone replacement therapy. AB - BACKGROUND: Dietary docosahexaenoic acid (DHA) has triacylglycerol-lowering potential and undergoes in vivo retroconversion to eicosapentaenoic acid (EPA) in humans. Hormone replacement therapy (HRT) influences circulating lipid concentrations and fatty acid metabolism. DHA supplementation has not been studied in postmenopausal women. OBJECTIVE: We studied the effects of supplementation with DHA (free of EPA) on the resulting elevation in EPA and on selected cardiovascular disease risk factors in postmenopausal women. DESIGN: Women receiving (n = 18) and not receiving (n = 14) HRT completed a randomized, double-blind, placebo-controlled crossover trial with a DHA supplement (2.8 g DHA/d). A washout period of > or =6 wk divided the two 28-d intervention periods. Fasting blood samples were collected for analysis. RESULTS: In all women, DHA supplementation was associated with significant changes (P < 0.05), including 20% lower serum triacylglycerol concentrations, 8% higher HDL-cholesterol concentrations, a 28% lower overall ratio of serum triacylglycerol to HDL cholesterol, and a 7% decrease in resting heart rate. DHA supplementation resulted in a 45% lower net increase (P = 0.02) in EPA and a 42% lower (P = 0.0028) estimated percentage retroconversion of DHA to EPA [DeltaEPA/(DeltaEPA + DeltaDHA) x 100] in women receiving than in those not receiving HRT. CONCLUSION: With DHA supplementation, the accumulation of EPA in serum phospholipids is significantly attenuated in postmenopausal women receiving HRT compared with that in women not receiving HRT. DHA supplementation can also favorably influence selected cardiovascular disease risk factors in postmenopausal women. PMID- 15113714 TI - Increased consumption of refined carbohydrates and the epidemic of type 2 diabetes in the United States: an ecologic assessment. AB - BACKGROUND: Type 2 diabetes is an epidemic that is affecting an ever-increasing proportion of the US population. Although consumption of refined carbohydrates has increased and is thought to be related to the increased risk of type 2 diabetes, the ecologic effect of changes in the quality of carbohydrates in the food supply on the risk of type 2 diabetes remains to be quantified. OBJECTIVE: The objective was to examine the correlation between consumption of refined carbohydrates and the prevalence of type 2 diabetes in the United States. METHODS: In this ecologic correlation study, the per capita nutrient consumption in the United States between 1909 and 1997 obtained from the US Department of Agriculture was compared with the prevalence of type 2 diabetes obtained from the Centers for Disease Control and Prevention. RESULTS: In a univariate analysis, a significant correlation with diabetes prevalence was observed for dietary fat (r = 0.84, P < 0.001), carbohydrate (r = 0.55, P < 0.001), protein (r = 0.71, P < 0.001), fiber (r = 0.16, P = 0.03), corn syrup (r = 0.83, P < 0.001), and total energy (r = 0.75, P < 0.001) intakes. In a multivariate nutrient-density model, in which total energy intake was accounted for, corn syrup was positively associated with the prevalence of type 2 diabetes (beta = 0.0132, P = 0.038). Fiber (beta = -13.86, P < 0.01) was negatively associated with the prevalence of type 2 diabetes. In contrast, protein (P = 0.084) and fat (P = 0.79) were not associated with the prevalence of type 2 diabetes when total energy was controlled for. CONCLUSIONS: Increasing intakes of refined carbohydrate (corn syrup) concomitant with decreasing intakes of fiber paralleled the upward trend in the prevalence of type 2 diabetes observed in the United States during the 20th century. PMID- 15113715 TI - Body-composition differences between African American and white women: relation to resting energy requirements. AB - BACKGROUND: Body composition differs between African American (AA) and white women, and the resting metabolic rate (RMR) is likely to be lower in AA women than in white women. OBJECTIVE: We tested 2 hypotheses: that AA women have a greater proportion of low-metabolic-rate skeletal muscle (SM) and bone than do white women and that between-race musculoskeletal differences are a function of body weight. DESIGN: Hypothesis 1 was tested by comparing SM, bone, adipose tissue, and high-metabolic-rate residual mass across 22 pairs of matched AA and white women. Magnetic resonance imaging and dual-energy X-ray absorptiometry were used to partition weight into 4 components, and RMR was both calculated from tissue-organ mass and measured. Hypothesis 2 was evaluated by measuring SM, bone, fat, and residual mass in 521 AA and white women with the use of dual-energy X ray absorptiometry alone. RESULTS: Hypothesis 1: AA women had greater SM ( +/- SD group difference: 1.52 +/- 2.48 kg; P < 0.01) and musculoskeletal mass (1.72 +/- 2.66 kg; P < 0.01) than did white women. RMR calculated from body composition and measured RMR did not differ; RMR estimated by both approaches tended to be lower (approximately 160 kJ/d) in AA women than in white women. Hypothesis 2: SM was significantly correlated with weight, height, age, and race x weight interaction; greater SM in the AA women was a function of body weight. CONCLUSIONS: Lower RMRs in AA women than in white women are related to corresponding differences in the proportions of heat-producing tissues and organs, and these race-related body composition differences increase as a function of body weight. PMID- 15113716 TI - Effect of feeding malnourished patients for 1 mo on mitochondrial complex I activity and nutritional assessment measurements. AB - BACKGROUND: We showed previously that the activity of complex I (the first enzyme of the electron transport chain) in peripheral blood mononuclear cells decreases with malnutrition and increases to a subnormal value after 1 wk of refeeding, but the traditional markers of nutritional status do not do so. OBJECTIVE: The aim of this study was to ascertain whether a period of nutritional intervention longer than 1 wk would normalize complex I activity and traditional markers of nutritional status. DESIGN: Fifteen malnourished patients (7 women and 8 men) with > or =10% body weight loss over the previous 6 mo were studied on the day of their admission to hospital and 7, 14 and 30 d after the beginning of nutritional support. Complex I activity in peripheral blood mononuclear cells, weight, height, body composition, body water compartments, dietary intake, and serum albumin concentrations were measured on each occasion. The results before and during nutritional intervention were compared with values obtained in 30 healthy volunteers (17 women and 13 men). RESULTS: Complex I activity increased significantly after the first week of refeeding (P < 0.001) and reached a normal value after 1 mo of nutritional supplementation. Among the classic markers of nutritional status, only the ratio of extracellular water to intracellular water tended to decrease over the refeeding period. CONCLUSION: Complex I activity increases rapidly and is normalized by refeeding at a time when other markers of nutritional status do not change significantly. PMID- 15113717 TI - Psychosocial predictors of energy underreporting in a large doubly labeled water study. AB - BACKGROUND: Underreporting of energy intake is associated with self-reported diet measures and appears to be selective according to personal characteristics. Doubly labeled water is an unbiased reference biomarker for energy intake that may be used to assess underreporting. OBJECTIVE: Our objective was to determine which factors are associated with underreporting of energy intake on food frequency questionnaires (FFQs) and 24-h dietary recalls (24HRs). DESIGN: The study participants were 484 men and women aged 40-69 y who resided in Montgomery County, MD. Using the doubly labeled water method to measure total energy expenditure, we considered numerous psychosocial, lifestyle, and sociodemographic factors in multiple logistic regression models for prediction of the probability of underreporting on the FFQ and 24HR. RESULTS: In the FFQ models, fear of negative evaluation, weight-loss history, and percentage of energy from fat were the best predictors of underreporting in women (R(2) = 0.09); body mass index, comparison of activity level with that of others of the same sex and age, and eating frequency were the best predictors in men (R(2) = 0.10). In the 24HR models, social desirability, fear of negative evaluation, body mass index, percentage of energy from fat, usual activity, and variability in number of meals per day were the best predictors of underreporting in women (R(2) = 0.22); social desirability, dietary restraint, body mass index, eating frequency, dieting history, and education were the best predictors in men (R(2) = 0.25). CONCLUSION: Although the final models were significantly related to underreporting on both the FFQ and the 24HR, the amount of variation explained by these models was relatively low, especially for the FFQ. PMID- 15113718 TI - Breakfast cereal fortified with folic acid, vitamin B-6, and vitamin B-12 increases vitamin concentrations and reduces homocysteine concentrations: a randomized trial. AB - BACKGROUND: High homocysteine and low B vitamin concentrations have been linked to the risk of vascular disease, stroke, and dementia and are relatively common in older adults. OBJECTIVE: We assessed the effect of breakfast cereal fortified with folic acid, vitamin B-6, and vitamin B-12 on vitamin and homocysteine status. DESIGN: A randomized, double-blind trial was conducted in 189 volunteers aged 50-85 y. The subjects had no history of hypertension, anemia, asthma, cancer, or cardiovascular or digestive disease and did not regularly consume multiple or B vitamin supplements or highly fortified breakfast cereal. Subjects were randomly assigned to consume 1 cup (0.24 L) breakfast cereal fortified with 440 microg folic acid, 1.8 mg vitamin B-6, and 4.8 microg vitamin B-12 or placebo cereal for 12 wk. Blood was drawn at 0, 2, 12, and 14 wk. Methionine-loading tests were conducted at baseline and week 14. RESULTS: Final baseline-adjusted plasma homocysteine concentrations were significantly lower and B vitamin concentrations were significantly higher in the treatment group than in the placebo group (P < 0.001). The percentage of subjects with plasma folate concentrations < 11 nmol/L decreased from 2% to 0%, with vitamin B-12 concentrations < 185 pmol/L from 9% to 3%, with vitamin B-6 concentrations < 20 nmol/L from 6% to 2%, and with homocysteine concentrations > 10.4 micromol/L (women) or > 11.4 micromol/L (men) from 6.4% to 1.6%. The percentage of control subjects with values beyond these cutoff points remained nearly constant or increased. CONCLUSIONS: In this relatively healthy group of volunteers, consumption of 1 cup fortified breakfast cereal daily significantly increased B vitamin and decreased homocysteine concentrations, including post-methionine-load homocysteine concentrations. PMID- 15113719 TI - Changes in lifestyle and plasma total homocysteine: the Hordaland Homocysteine Study. AB - BACKGROUND: Elevated plasma concentrations of total homocysteine (tHcy) are a risk factor for cardiovascular disease. tHcy is a marker of folate and cobalamin deficiencies and is also related to several lifestyle factors. OBJECTIVE: We examined whether changes in lifestyle influence tHcy over time. DESIGN: A population-based, prospective study was conducted in 7031 subjects from western Norway who constituted 2 age groups (41-42 and 65-67 y) at baseline (1992-1993). The subjects were reinvestigated in 1997-1999 ( follow-up: 6 y). RESULTS: During follow-up, median tHcy concentrations decreased 0.10 (25th and 75th percentiles: 1.24, 1.00) micromol/L in the younger subjects and increased 0.39 (25th and 75th percentiles: -0.99, 1.79) micromol/L in the older subjects. Changes in plasma vitamin status and vitamin supplement use were the strongest determinants of changes in tHcy over time. Each unit increase in plasma folate (nmol/L) and vitamin B-12 (pmol/L) was associated with reductions in tHcy concentrations of 0.2 and 0.1 micromol/L, respectively. Among the younger and older age groups, those who started to take vitamin supplements during follow-up had significant reductions in tHcy concentrations of 0.42 (95% CI: -0.65, -0.20) and 0.41 (-0.78, -0.03) micromol/L, respectively. In the younger subjects who quit smoking, tHcy concentrations decreased 0.54 (-0.91, -0.16) micromol/L. Weight changes were inversely related to tHcy. Both baseline history of cardiovascular disease or hypertension and cardiovascular events during follow-up were significantly associated with changes in tHcy. CONCLUSIONS: Changes in lifestyle factors over time influence tHcy concentrations. These changes are modest when compared with the strong associations between tHcy and lifestyle factors in cross-sectional studies. PMID- 15113720 TI - Hypovitaminosis D is associated with insulin resistance and beta cell dysfunction. AB - BACKGROUND: Although the role of vitamin D in type 2 diabetes is well recognized, its relation to glucose metabolism is not well studied. OBJECTIVE: We investigated the relation of 25-hydroxyvitamin D [25(OH)D] concentrations to insulin sensitivity and beta cell function. DESIGN: We enrolled 126 healthy, glucose-tolerant subjects living in California. Insulin sensitivity index (ISI) and first- and second-phase insulin responses (1stIR and 2ndIR) were assessed by using a hyperglycemic clamp. RESULTS: Univariate regression analyses showed that 25(OH)D concentration was positively correlated with ISI (P < 0.0001) and negatively correlated with 1stIR (P = 0.0045) and 2ndIR (P < 0.0001). Multiple regression analyses confirmed an independent correlation between 25(OH)D concentration and ISI (P = 0.0007). No independent correlation was observed between 25(OH)D concentration and 1stIR or 2ndIR. However, an independent negative relation of 25(OH)D concentration with plasma glucose concentration was observed at fasting (P = 0.0258), 60 min (P = 0.0011), 90 min (P = 0.0011), and 120 min (P = 0.0007) during the oral-glucose-tolerance test. Subjects with hypovitaminosis D (<20 ng/mL) had a greater prevalence of components of metabolic syndrome than did subjects without hypovitaminosis D (30% compared with 11%; P = 0.0076). CONCLUSIONS: The data show a positive correlation of 25(OH)D concentration with insulin sensitivity and a negative effect of hypovitaminosis D on beta cell function. Subjects with hypovitaminosis D are at higher risk of insulin resistance and the metabolic syndrome. Further studies are required to explore the underlying mechanisms. PMID- 15113721 TI - Randomized controlled trial of prenatal zinc supplementation and fetal bone growth. AB - BACKGROUND: Maternal zinc deficiency is relatively common in developing countries, but its consequences for fetal growth are not established. OBJECTIVE: The goal was to examine whether improvement in maternal gestational zinc status is positively associated with fetal growth as assessed by ultrasonography. DESIGN: We conducted a double-masked, randomized trial among 242 pregnant Peruvian women in an impoverished shantytown in Lima, Peru. At 10-16 wk of gestation, the women were randomly assigned to receive daily supplements containing 60 mg Fe and 250 microg folic acid, with or without 25 mg Zn. We measured fetal head circumference, biparietal diameter, abdominal circumference, and femur diaphysis length at 20, 24, 28, 32, 36, and 38 wk of gestation. Fetal measures were analyzed longitudinally to evaluate differences in trends of fetal growth by supplement type, and within-subject correlations were taken into account. RESULTS: Femur diaphysis length was greater in fetuses whose mothers received zinc supplements (P < 0.05), and the difference tended to increase with gestational age. No significant differences by supplement type were observed for the other anatomical sites measured. CONCLUSIONS: The observed positive effect of prenatal zinc on fetal femur diaphysis length is consistent with the results of experimental studies in animals and in vitro. The supplementation effect represents an upward shift in mean femur diaphysis length at term of about one quarter of the reference SD. These findings suggest the potential importance of maternal zinc status for fetal bone growth in humans and illustrate the value of ultrasonography for evaluating the effect of prenatal nutritional interventions on components of fetal growth. PMID- 15113722 TI - Splanchnic bed metabolism of glucose in preterm neonates. AB - BACKGROUND: Glucose is a major oxidative substrate for intestinal energy generation in neonatal animals; however, few data in preterm infants are available. Early administration of enteral nutrition, including glucose, may be an effective strategy to support intestinal adaptation to extrauterine life in preterm neonates. OBJECTIVE: The purpose of the present study was to quantify the first-pass uptake and oxidation of glucose by the splanchnic tissues (intestine and liver) in human neonates. DESIGN: Eight preterm infants [birth weight ( +/- SD): 1.19 +/- 0.22 kg, gestational age: 29 +/- 1 wk] were studied while they received 2 different enteral intakes (A: 40% enteral, 60% parenteral, total glucose intake = 7.5 +/- 0.5 mg. kg(-1). min(-1), and B: 100% enteral, total glucose intake = 7.8 +/- 0.4 mg. kg(-1). min(-1)). Splanchnic and whole-body glucose kinetics were measured by use of dual-tracer techniques. RESULTS: During both feeding periods, approximately one-third of dietary glucose intake was utilized during the first pass by the splanchnic tissues. More than three quarters of this utilized glucose was oxidized in both periods (79 +/- 36% with A and 84 +/- 45% with B). Whole-body glucose oxidation was substantial under both circumstances: 72 +/- 5% and 77% +/- 6% of the glucose flux was oxidized during partial (A) and full (B) enteral feeding, respectively. CONCLUSIONS: Approximately one-third of dietary glucose is utilized during the first pass by the splanchnic tissues, irrespective of the dietary intake. Most of the utilized glucose is used for energy generation. PMID- 15113723 TI - Detection of overweight and obesity in a national sample of 6-12-y-old Swiss children: accuracy and validity of reference values for body mass index from the US Centers for Disease Control and Prevention and the International Obesity Task Force. AB - BACKGROUND: For defining overweight in children, reference values for body mass index (BMI) are available from the US Centers for Disease Control and Prevention (CDC) and the International Obesity Task Force (IOTF). However, these 2 sets of reference criteria differ, and their accuracy in classifying adiposity has not yet been validated in most countries. OBJECTIVE: We compared BMI criteria from the IOTF and the CDC with percentage of body fat (%BF) from multisite skinfold thicknesses (SFTs) for identification of overweight in 6-12-y-old Swiss children. DESIGN: In a representative sample (n = 2431), weight, height, and 4 SFTs were measured. Regression and receiver operating characteristic (ROC) curves were used to evaluate BMI as an indicator of adiposity. RESULTS: BMI and %BF were well correlated (r(2) = 0.74), and the areas under the ROC curves for overweight and obesity were 0.956-0.992. The sensitivity and specificity of the IOTF and CDC overweight criteria and of the CDC obesity criteria were high. The sensitivity of the IOTF obesity criteria was only 48% and 62% in boys and girls, respectively. Overall, the performance of the CDC criteria was superior. With the use of the CDC criteria, the prevalence of overweight in girls and boys was 19.1% and 20.3%, respectively. CONCLUSIONS: BMI is an excellent proxy measure of adiposity in 6-12 y-old children. In Swiss children, both BMI criteria accurately predict overweight, but the sensitivity of the IOTF obesity criteria is poor. They failed to detect one-half of the children identified as obese on the basis of %BF from SFTs. PMID- 15113724 TI - Familial aggregation of energy intake in children. AB - BACKGROUND: Uncompensated overnutrition promotes obesity, but the controls of children's eating behavior are poorly understood. Insights may be achieved by testing whether the eating patterns of children are associated with demographic variables or whether they aggregate among family members. OBJECTIVE: We tested whether children's total energy intake and macronutrient intake and their ability to compensate for earlier energy intake were associated with sociodemographic variables and anthropometric indexes. We also tested whether these behavioral traits aggregate among siblings. DESIGN: Thirty-two sibling pairs aged 3-7 y consumed a multi-item lunch preceded by a low-energy (12.55 kJ) or high-energy (627.60 kJ) preload drink. Mixed-models regression tested the associations between children's energy intake, demographic variables, and anthropometric measures. An intraclass correlation coefficient quantified the family correlation of the measures of children's eating. RESULTS: Children consumed significantly more total energy after consuming the low-energy preload ( +/- SD: 2237.39 +/- 1176.45 kJ) than after consuming the high-energy preload (1601.18 +/- 930.65 kJ). Compensation ability was unrelated to the children's age, sex, or ethnicity. Total energy and macronutrient intake, but not compensation propensity, were associated among siblings. CONCLUSIONS: The familial association of total energy and macronutrient intakes, independent of anthropometric measures, suggests genetic or home environmental influences specific to these behaviors. Short-term energy compensation, although very accurate within this sample, showed no significant familial correlation. PMID- 15113725 TI - Body movement and physical activity energy expenditure in children and adolescents: how to adjust for differences in body size and age. AB - BACKGROUND: Physical activity data in children and adolescents who differ in body size and age are influenced by whether physical activity is expressed in terms of body movement or energy expenditure. OBJECTIVE: We examined whether physical activity expressed as body movement (ie, accelerometer counts) differs from physical activity energy expenditure (PAEE) as a function of body size and age. DESIGN: This was a cross-sectional study in children [n = 26; (+/-SD) age: 9.6 +/ 0.3 y] and adolescents (n = 25; age: 17.6 +/- 1.5 y) in which body movement and total energy expenditure (TEE) were simultaneously measured with the use of accelerometry and the doubly labeled water method, respectively. PAEE was expressed as 1) unadjusted PAEE [TEE minus resting energy expenditure (REE); in MJ/d], 2) PAEE adjusted for body weight (BW) (PAEE. kg(-1). d(-1)), 3) PAEE adjusted for fat-free mass (FFM) (PAEE. kg FFM(-1). d(-1)), and 4) the physical activity level (PAL = TEE/REE). RESULTS: Body movement was significantly higher (P = 0.03) in children than in adolescents. Similarly, when PAEE was normalized for differences in BW or FFM, it was significantly higher in children than in adolescents (P = 0.03). In contrast, unadjusted PAEE and PAL were significantly higher in adolescents (P < 0.01). CONCLUSIONS: PAEE should be normalized for BW or FFM for comparison of physical activity between children and adolescents who differ in body size and age. Adjusting PAEE for FFM removes the confounding effect of sex, and therefore FFM may be the most appropriate body-composition variable for normalization of PAEE. Unadjusted PAEE and PAL depend on body size. PMID- 15113726 TI - Intake of specific carotenoids and essential fatty acids and breast cancer risk in Montreal, Canada. AB - BACKGROUND: Evidence from previous investigations into the possible role of dietary and serum carotenoid concentrations in the etiology of breast cancer is inconsistent. No study has examined the combined effect of carotenoids and essential fatty acids on the risk of breast cancer. OBJECTIVE: The objective was to assess the possible association between specific and total carotenoids and breast cancer risk and to evaluate the effect modification by diet-related fatty acids and lifestyle factors in the development of breast cancer. DESIGN: A population-based case-control study involving 414 incident cases and 429 controls was conducted in French Canadians in Montreal. Dietary intake was estimated with the use of a validated food-frequency questionnaire in face-to-face interviews. RESULTS: No significant association was apparent between any of the individual or total carotenoids and the risk of breast cancer after adjustment for major underlying determinants of breast cancer. In premenopausal women who ever smoked, an increased risk was related to alpha-carotene [odds ratio (OR) for the upper relative to the lowest quartiles of intake: 2.40; 95% CI: 0.90, 6.41; P for trend = 0.046]. Conversely, a reduced risk was related to beta-carotene (OR: 0.57; 95% CI: 0.26, 1.24; P for trend = 0.05) in women who never used hormone replacement therapy. In postmenopausal women, total carotenoids were positively associated with breast cancer risk in those with a high arachidonic acid intake (OR: 1.92; 95% CI: 0.93, 3.94; P = 0.028 for trend) and inversely associated in those with a high docosahexaenoic acid intake (OR: 0.52; 95% CI: 0.25, 1.07; P for trend = 0.054). CONCLUSION: These findings suggest that the combined high intake of total carotenoids and docosahexaenoic acid may reduce the risk of breast cancer. PMID- 15113727 TI - Relation between intake of vitamins C and E and risk of diabetic retinopathy in the Atherosclerosis Risk in Communities Study. AB - BACKGROUND: The potential protective effect of vitamins C and E against the development of diabetic retinopathy has not been thoroughly evaluated in epidemiologic studies. OBJECTIVE: The objective was to study the association between prevalent diabetic retinopathy and intake of vitamins C and E in participants of the Atherosclerosis Risk in Communities Study. DESIGN: A total of 1353 subjects with type 2 diabetes diagnosed between 1993 and 1995 or before were included. Nutrient intake was assessed with a food-frequency and supplement questionnaire administered between 1987-1989 and 1993-1995. Prevalent retinopathy (n = 224) was determined in 1993-1995 from graded fundus photographs. RESULTS: No association of retinopathy with intake of vitamin C or E from food alone or from food and supplements combined was observed. The odds ratios and 95% CIs for retinopathy for quartile 4 compared with quartile 1 of vitamins C and E intakes from food and supplements combined were 1.1 (0.7, 1.9) and 1.3 (0.8, 2.2), respectively, after adjustment for diabetes treatment and serum glucose. There was a significant interaction of the observed relations with serum glucose concentration (P < 0.05). Additionally, a decreased odds of retinopathy was found among users (reported use > or =3 y before 1993-1995) of vitamin C or E supplements or multisupplements compared with reported use of no supplements: 0.5 (0.3, 0.8), 0.5 (0.2, 0.8), and 0.4 (0.2, 0.9), respectively. CONCLUSION: No significant overall associations were observed between risk of retinopathy and intake of major dietary antioxidants. The observed association between risk of retinopathy and supplement use may reflect nondietary factors or a possible benefit of supplementation. PMID- 15113728 TI - Sarcopenia and increased adipose tissue infiltration of muscle in elderly African American women. AB - BACKGROUND: Aging is associated with metabolic, physiologic, and functional impairments, in part through age-related changes in body composition. During the later adult years, skeletal muscle mass decreases and body fat becomes centralized. OBJECTIVE: The goal of the study was to investigate body composition over time ( +/- SD: 2.04 +/- 0.6 y) in healthy, ambulatory, elderly African American women. The hypothesis that a reduction in total-body skeletal muscle (SM) and increases in visceral adipose tissue (VAT), subcutaneous adipose tissue (SAT), and intermuscular adipose tissue (IMAT) are ongoing in healthy, weight stable elderly was tested. DESIGN: The study was a longitudinal evaluation of 26 women (age at baseline: 75.5 +/- 5.1 y) with a body mass index (in kg/m(2)) of 27.0 +/- 4.0. Body composition was measured by using whole-body magnetic resonance imaging for the quantification of SM, total adipose tissue (TAT), VAT, SAT, and IMAT. RESULTS: SM (P < 0.001) and bone (P < 0.05) masses decreased, and regional analyses showed a decrease in dual-energy X-ray absorptiometry-derived leg SM (P < 0.05). VAT (P = 0.011) and IMAT (P < 0.001) increased. No changes occurred in TAT (P = 0.45), SAT (P = 0.96), physical function, or food intake. CONCLUSION: These data show an age-related remodeling of body composition with reductions in SM and corresponding increases in VAT and IMAT. Changes in the previously unstudied depot of IMAT may be involved in the deterioration of metabolic values frequently observed during aging. PMID- 15113729 TI - A genome-wide linkage scan for dietary energy and nutrient intakes: the Health, Risk Factors, Exercise Training, and Genetics (HERITAGE) Family Study. AB - BACKGROUND: A poor diet is a risk factor for chronic diseases such as obesity, cardiovascular disease, hypertension, and some cancers. Twin and family studies suggest that genetic factors potentially influence energy and nutrient intakes. OBJECTIVE: We sought to identify genomic regions harboring genes affecting total energy, carbohydrate, protein, and fat intakes. DESIGN: We performed a genomic scan in 347 white sibling pairs and 99 black sibling pairs. Dietary energy and nutrient intakes were assessed by using Willett's food-frequency questionnaire. Single-point and multipoint Haseman-Elston regression techniques were used to test for linkage. These subjects were part of the Health, Risk Factors, Exercise Training, and Genetics (HERITAGE) Family Study, a multicenter project undertaken by 5 laboratories. RESULTS: In the whites, the strongest evidence of linkage appeared for dietary energy and nutrient intakes on chromosomes 1p21.2 (P = 0.0002) and 20q13.13 (P = 0.00007), and that for fat intake appeared on chromosome 12q14.1 (P = 0.0013). The linkage evidence on chromosomes 1 and 20 related to total energy intake rather than to the intake of specific macronutrients. In the blacks, promising linkages for macronutrient intakes occurred on chromosomes 12q23-q24.21, 1q32.1, and 7q11.1. Several potential candidate genes are encoded in and around the linkage regions on chromosomes 1p21.2, 12q14.1, and 20q13.13. CONCLUSIONS: These are the first reported human quantitative trait loci for dietary energy and macronutrient intakes. Further study may refine these quantitative trait loci to identify potential candidate genes for energy and specific macronutrient intakes that would be amenable to more detailed molecular studies. PMID- 15113731 TI - Chemical validation of X-ray absorptiometry. PMID- 15113732 TI - Is hyperleptinemia involved in the development of age-related lens opacities? PMID- 15113733 TI - Vitamin D and type 1 diabetes. PMID- 15113736 TI - The weight debate: balancing food composition and physical activity. Preface. PMID- 15113737 TI - Is a calorie a calorie? AB - The aim of this review was to evaluate data regarding potential thermodynamic mechanisms for increased rates of weight loss in subjects consuming diets high in protein and/or low in carbohydrate. Studies that compared weight loss and energy expenditure in adults consuming diets high in protein and/or low in carbohydrate with those in adults consuming diets low in fat were reviewed. In addition, studies that measured the metabolizable energy of proteins, fats, and carbohydrates were reviewed. Diets high in protein and/or low in carbohydrate produced an approximately equal to 2.5-kg greater weight loss after 12 wk of treatment. Neither macronutrient-specific differences in the availability of dietary energy nor changes in energy expenditure could explain these differences in weight loss. Thermodynamics dictate that a calorie is a calorie regardless of the macronutrient composition of the diet. Further research on differences in the composition of weight loss and on the influence of satiety on compliance with energy-restricted diets is needed to explain the observed increase in weight loss with diets high in protein and/or low in carbohydrate. PMID- 15113738 TI - Role of calcium and dairy products in energy partitioning and weight management. AB - Dietary calcium plays a pivotal role in the regulation of energy metabolism because high-calcium diets attenuate adipocyte lipid accretion and weight gain during the overconsumption of an energy-dense diet and increase lipolysis and preserve thermogenesis during caloric restriction, which thereby markedly accelerates weight loss. Intracellular Ca(2+) plays a key regulatory role in adipocyte lipid metabolism and triacylglycerol storage; increased intracellular Ca(2+) results in the stimulation of lipogenic gene expression and lipogenesis and the suppression of lipolysis, which results in increased lipid filling and increased adiposity. Moreover, the increased calcitriol produced in response to low-calcium diets stimulates adipocyte Ca(2+) influx and, consequently, promotes adiposity, whereas higher-calcium diets inhibit lipogenesis, inhibit diet-induced obesity in mice, and promote lipolysis, lipid oxidation, and thermogenesis. Notably, dairy sources of calcium markedly attenuate weight and fat gain and accelerate fat loss to a greater degree than do supplemental sources of calcium. This augmented effect of dairy products relative to supplemental calcium is likely due to additional bioactive compounds, including the angiotensin converting enzyme inhibitors and the rich concentration of branched-chain amino acids in whey, which act synergistically with calcium to attenuate adiposity. These concepts are confirmed by epidemiologic data and recent clinical trials, which indicate that diets that include > or =3 daily servings of dairy products result in significant reductions in adipose tissue mass in obese humans in the absence of caloric restriction and markedly accelerate weight and body fat loss secondary to caloric restriction compared with diets low in dairy products. These data indicate an important role for dairy products in both the prevention and treatment of obesity. PMID- 15113739 TI - The evolution of physical activity recommendations: how much is enough? AB - Physical inactivity is a major public health problem, and compelling evidence suggests that it is a contributing factor in several chronic diseases and conditions. Recognition of the health and functional hazards of a sedentary way of life has led numerous groups to promulgate public health recommendations for physical activity. In this report, we review the evolution of physical activity recommendations, discuss reasons for differences in the recommendations, and provide a summary recommendation in an attempt to harmonize existing differences. Current public health recommendations for physical activity are for 30 min of moderate-intensity activity each day, which provides substantial benefits across a broad range of health outcomes for sedentary adults. This dose of exercise may be insufficient to prevent unhealthful weight gain for some persons who may need additional exercise or caloric restriction to minimize the likelihood of further weight gain. Persons who get 30 min of moderate-intensity exercise per day are likely to achieve additional health benefits if they exercise more. In addition to aerobic exercise, people should engage in resistance training and flexibility exercises at least twice a week, which will promote the maintenance of lean body mass, improvements in muscular strength and endurance, and preservation of function, all of which enable long-term participation in regular physical activity and promote quality of life. PMID- 15113740 TI - Chronicle of the Institute of Medicine physical activity recommendation: how a physical activity recommendation came to be among dietary recommendations. AB - Under a contract from the US Department of Health and Human Services, a multidisciplinary expert panel was appointed to review "the scientific literature regarding macronutrients and energy and develop estimates of daily intake that are compatible with good nutrition throughout the life span and that may decrease the risk of chronic disease." Within the overall context of the charge, the panel sought to quantify rates and components of daily energy expenditure in healthy adults with body mass indexes (in kg/m(2)) of 18.5-25, in growing children (in the 5th-85th percentiles of weight-for-length), and in pregnant and lactating women. The recommendation for adults became the daily energy intake necessary to cover total daily energy expenditure (TEE). For special cases, dietary macronutrients and energy to support child growth and pregnancy and lactation by women were considered. TEE was based on the results of doubly labeled water studies, and the TEE results were presented in units of physical activity level (PAL = TEE/BEE) and DeltaPAL, where BEE is the basal rate of energy expenditure extrapolated to 24 h. Most adults (66%) maintaining a BMI in the healthful range had PAL values >1.6, or the equivalent of > or =60 min of physical activity of moderate intensity each day. Hence, on the basis of the doubly labeled water data and the results of epidemiologic studies, the physical activity recommendation for adults was judged to be 60 min/d. The recommendation for children was for a minimum of 60 min/d. In conclusion, dietary and physical activity recommendations for healthful living are inextricably intertwined. Adequate physical activity provides protection against chronic diseases and helps to balance energy expenditure and intake. PMID- 15113741 TI - Pollination ecology of four epiphytic orchids of New Zealand. AB - BACKGROUND AND AIMS: In New Zealand epiphytic orchids are represented by four genera and eight species. The genera Earina (three species) and Winika (one species) are the most conspicuous and widespread. These are likely to be some of the southernmost distributed genera of epiphytic orchids in the world. METHODS: To identify the pollination strategies that have evolved in these orchids, hand pollination treatments were done and floral visitors were observed in several wild populations at two areas of southern North Island (approx. 40 degrees S). Pollen:ovule ratio and osmophores were also studied and the total carbohydrate content of the nectar produced by each species was measured. KEY RESULTS: Earina autumnalis and Earina mucronata are self-compatible, whereas Earina aestivalis and Winika cunninghamii appear to be partially self-incompatible. All four orchids are incapable of autonomous selfing and therefore completely dependent on pollinators to set fruits. Floral visitors observed in the genus Earina belong to Diptera, Coleoptera and Hymenoptera and to Diptera and Hymenoptera in W. cunninghamii. CONCLUSIONS: Contrary to many epiphytic orchids in the tropics, the orchid-pollinator relationship in these orchids is unspecialized and flowers are visited by a wide range of insects. Putative pollinators are flies of the families Bibionidae, Calliphoridae, Syrphidae and Tachinidae. All four orchids display anthecological adaptations to a myophilous pollination system such as simple flowers, well-exposed reproductive structures, easily accessed nectar and high pollen : ovule ratios. PMID- 15113742 TI - Maturation of the Na+/H+ antiporter (NHE3) in the proximal tubule of the hypothyroid adrenalectomized rat. AB - In previous studies examining the role of glucocorticoids and thyroid hormone on the maturation of the Na(+)/H(+) antiporter (NHE3), we found attenuation in the maturational increase in proximal tubule apical Na(+)/H(+) antiporter activity but no change in NHE3 mRNA abundance in either glucocorticoid-deficient or hypothyroid rats. In addition, prevention of the maturational increase in either hormone failed to totally prevent the maturational increase in Na(+)/H(+) antiporter activity. We hypothesized that one hormone played a compensatory role when the other was deficient. The present study examined whether combined deficiency of thyroid and glucocorticoid hormones would completely prevent the maturation of the Na(+)/H(+) antiporter. Adrenalectomy was performed in 9-day-old hypothyroid Sprague-Dawley rats, a time before the normal postnatal maturational increase in these hormones occurs. Nine- and 30-day-old adrenalectomized (ADX), hypothyroid rats had comparable NHE3 mRNA abundance, which was 5- to 10-fold less than 30-day-old ADX, hypothyroid rats that received corticosterone-thyroxine replacement and 30-day-old sham control rats (P < 0.05). Brush-border membrane NHE3 protein abundance was comparable in 9- and 30-day-old ADX, hypothyroid groups and approximately 20-fold lower than both the 30-day replacement and 30 day sham groups (P < 0.05). Similarly, the replacement and sham groups had higher sodium-dependent proton secretion than 9- and 30-day-old ADX, hypothyroid groups (P < 0.05). We conclude that combined deficiency of both hormones totally prevents the maturational increase in NHE3 mRNA and protein abundance and Na(+)/H(+) antiporter activity. PMID- 15113743 TI - The role of bladder-to-urethral reflexes in urinary continence mechanisms in rats. AB - Urethral closure mechanisms during passive increments in intravesicular pressure (P(ves)) were investigated using microtip transducer catheters in urethane anesthetized female rats. After a block of reflex bladder contractions by spinal cord transection at T8-T9, abruptly raising P(ves) to 20, 40, or 60 cmH(2)O for 2 min induced a bladder pressure-dependent contractile response in a restricted portion of the middle urethra (12.5-15 mm from the urethral orifice) that was abolished by cutting the pelvic nerves bilaterally. In pelvic nerve-intact rats, the bilateral transection of either the pudendal nerves, the nerves to the iliococcygeous/pubococcygeous muscles, or the hypogastric nerves significantly reduced (49-74%) the urethral reflex response induced by passive P(ves) increases, and combined transection of these three sets of nerves totally abolished the urethra-closing responses. In spinal cord-intact rats, similar urethral contractile responses were elicited during P(ves) elevation (20 or 40 cmH(2)O) and were also eliminated by bilateral pelvic nerve transection. After spinal cord and pelvic nerve transection, leak point pressures, defined as the pressure inducing fluid leakage from the urethral orifice during passive P(ves) elevation by either bladder pressure clamping in 2.5-cmH(2)O steps or direct compression of the bladder, were significantly lowered by 30-35% compared with sham-operated (spinal cord-transected and pelvic nerve-intact) rats. These results indicate that 1) passive elevation of P(ves) can elicit pelvic afferent nerve-mediated contractile reflexes in the restricted portion of the urethra mediated by activation of sympathetic and somatic nerves and 2) bladder-to urethral reflexes induced by passive P(ves) elevation significantly contribute to the prevention of stress urinary incontinence. PMID- 15113744 TI - NHE3 Na+/H+ exchanger supports proximal tubular protein reabsorption in vivo. AB - Proximal tubular receptor-mediated endocytosis (RME) of filtered proteins prevents proteinuria. Pharmacological and genetic studies in cultured opossum kidney cells have shown that the apical Na(+)/H(+) exchanger isoform 3 (NHE3) supports RME by interference with endosomal pH homeostasis and endocytic fusion events. However, it is not known whether NHE3 also supports proximal tubular RME in vivo. We analyzed proximal tubular protein reabsorption by microinfusion experiments in rats and investigated renal protein excretion in NHE3 knockout (Nhe3 -/-) mice. Inhibition of NHE3 by EIPA or S-3226 reduced the fractional reabsorption of [(14)C]cytochrome c by approximately 50% during early proximal microinfusion. During early distal microinfusion, no protein reabsorption could be detected. Urinary protein excretion of Nhe3 -/- or heterozygous mutant mice was significantly higher compared with wild-type mice. SDS-PAGE analysis of urinary proteins revealed that Nhe3 -/- animals excreted proteins the size of albumin or smaller. Thus a reduction in NHE3 activity or abundance causes tubular proteinuria. These data show that NHE3 supports proximal tubular RME of filtered proteins in vivo. PMID- 15113745 TI - Stimulation of renin release by prostaglandin E2 is mediated by EP2 and EP4 receptors in mouse kidneys. AB - PGE(2) is a potent stimulator of renin release. So far, the contribution of each of the four PGE(2) receptor subtypes (EP(1)-EP(4)) in the regulation of renin release has not been characterized. Therefore, we investigated the effects PGE(2) on renin secretion rates (RSR) from isolated, perfused kidneys of EP(1)-/-, EP(2) /-, EP(3)-/-, EP(4)-/-, and wild-type mice. PGE(2) concentration dependently stimulated RSR from kidneys of all four knockout strains with a threshold concentration of 1 nM in EP(1)-/-, EP(2)-/-, EP(3)-/-, and wild-type mice, whereas the threshold concentration was shifted to 10 nM in EP(4)-/- mice. Moreover, the maximum stimulation of RSR by PGE(2) at 1 microM was significantly reduced in EP(4)-/- (12.8-fold of control) and EP(2)-/- (15.9-fold) compared with wild-type (20.7-fold), EP(1)-/- (23.8-fold), and EP(3)-/- (20.1-fold). In contrast, stimulation of RSR by either the loop diuretic bumetanide or the beta adrenoceptor agonist isoproterenol was similar in all strains. PGE(2) exerted a dual effect on renal vascular tone, inducing vasodilatation at low concentrations (1 nmol/) and vasoconstriction at higher concentrations (100 nmol/) in kidneys of wild-type mice. In kidneys of EP(2)-/- as well as EP(4)-/- mice, vasodilatation at low PGE(2) concentrations was prevented, whereas vasoconstriction at higher concentrations was augmented. In contrast, the vasodilatory component was pronounced in kidneys of EP(1) and EP(3) knockout mice, whereas in both genotypes the vasoconstriction at higher PGE(2) concentrations was markedly blunted. Our data provide evidence that PGE(2) stimulates renin release via activation of EP(2) and EP(4) receptors, whereas EP(1) and EP(3) receptors appear to be without functional relevance in juxtaglomerular cells. In contrast, all four receptor subtypes are involved in the control of renal vascular tone, EP(1) and EP(3) receptors increasing, and EP(2) as well as EP(4) receptors, decreasing it. PMID- 15113746 TI - Inhibition of apoptosis by Zn2+ in renal tubular cells following ATP depletion. AB - Apoptosis has been implicated in ischemic renal injury. Thus one strategy of renal protection is to antagonize apoptosis. However, apoptosis inhibitory approaches remain to be fully explored. Zn(2+) has long been implicated in apoptosis inhibition; but systematic analysis of the inhibitory effects of Zn(2+) is lacking. Moreover, whether Zn(2+) blocks renal cell apoptosis following ischemia is unknown. Here, we demonstrate that Zn(2+) is a potent apoptosis inhibitor in an in vitro model of renal cell ischemia. ATP depletion induced apoptosis in cultured renal tubular cells, which was accompanied by caspase activation. Zn(2+) at 10 microM inhibited both apoptosis and caspase activation, whereas Co(2+) was without effect. In ATP-depleted cells, Zn(2+) partially prevented Bax activation and cytochrome c release from mitochondria. In isolated cell cytosol, Zn(2+) blocked cytochrome c-stimulated caspase activation at low micromolar concentrations. In addition, Zn(2+) could directly antagonize the enzymatic activity of purified recombinant caspases. We conclude that Zn(2+) is a potent inhibitor of apoptosis in renal tubular cells following ATP depletion. Zn(2+) blocks apoptosis at multiple steps including Bax activation, cytochrome c release, apoptosome function, and caspase activation. PMID- 15113747 TI - Short-term peaks in glucose promote renal fibrogenesis independently of total glucose exposure. AB - Postprandial hyperglycemia is implicated as a risk factor predisposing to vascular complications. This study was designed to assess recurrent short-term increases in glucose on markers of renal fibrogenesis. Human renal cortical fibroblasts were exposed to fluctuating short-term (2 h) increases to 15 mM d glucose, three times a day over 72 h, on a background of 5 mM d-glucose. To determine whether observed changes were due to fluctuating osmolality, identical experiments were undertaken with cells exposed to l-glucose. Parallel experiments were performed in cells exposed to 5 mM d-glucose and constant exposure to either 15 or 7.5 mM d-glucose. Fluctuating d-glucose increased extracellular matrix, as measured by proline incorporation (P < 0.05), collagen IV (P < 0.005), and fibronectin production (P < 0.001), in association with increased tissue inhibitor of matrix metalloproteinase (MMP) (P < 0.05). Sustained exposure to 15 mM d-glucose increased fibronectin (P < 0.001), in association with increased MMP 2 (P = 0.01) and MMP-9 activity (P < 0.05), suggestive of a protective effect on collagen matrix accumulation. Transforming growth factor-beta(1) (TGF-beta(1)) mRNA was increased after short-term (90 min) exposure to 15 mM glucose (P < 0.05) and after 24-h exposure to 7.5 mM ? (P < 0.05). Normalization of TGF-beta(1) secretion occurred within 48 h of constant exposure to an elevated glucose. Fluctuating l-glucose also induced TGF-beta(1) mRNA and a profibrotic profile, however, to a lesser extent than observed with exposure to fluctuating d-glucose. The results suggest that exposure to fluctuating glucose concentrations increases renal interstitial fibrosis compared with stable elevations in d-glucose. The effects are, in part, due to the inherent osmotic changes. PMID- 15113748 TI - Annexin A2 heterotetramer: role in tight junction assembly. AB - The tight junction has been characterized as a domain of focal fusions of the exoplasmic leaflets of the lipid bilayers from adjacent epithelial cells. Approximating membranes to within fusion distance is a thermodynamically unfavorable process and requires the participation of membrane-bridging or fusion proteins. No known tight junction protein exhibits such activities. Annexin A2 (A2), in particular its heterotetramer (A2t), is known to form junctions between lipid bilayer structures through molecular bridging of their external leaflets. We demonstrate abundant A2 expression in Madin-Darby canine kidney II monolayers by two-dimensional gel electrophoresis. Confocal immunofluorescence microscopic analysis suggests the bulk of A2 is located along the apical and lateral plasma membrane in its tetrameric configuration, consisting of two A2 and two p11 (an 11-kDa calmodulin-related protein, S100A10) subunits. Immunocytochemistry and ultrastructural immunogold labeling demonstrate colocalization of the A2 subunit with bona fide tight junction proteins, zonula occludens-1, occludin, and claudin-1, at cell-cell contacts. The extracellular addition of a synthetic peptide, targeted to disrupt the binding between A2 and p11, completely aborts tight junction assembly in calcium chelation studies. We propose A2t as a member of a new class of tight junction proteins responsible for the long-observed convergence of adjacent exoplasmic lipid leaflets in tight junction assembly. PMID- 15113749 TI - Endothelin stimulates endothelial nitric oxide synthase expression in the thick ascending limb. AB - Endothelin-1 (ET-1) acutely inhibits NaCl reabsorption by the thick ascending limb (THAL) by activating the ET(B) receptor, stimulating endothelial nitric oxide synthase (eNOS), and releasing nitric oxide (NO). In nonrenal tissue, chronic exposure to ET-1 stimulates eNOS expression via the ET(B) receptor and activation of phosphatidylinositol 3-kinase (PI3K). We hypothesized that ET-1 increases eNOS expression in the THAL by binding to ET(B) receptors and stimulating PI3K. In primary cultures of medullary THALs treated for 24 h, eNOS expression increased by 36 +/- 18% with 0.01 nM ET-1, 123 +/- 30% with 0.1 nM (P < 0.05; n = 5), and 71 +/- 30% with 1 nM, whereas 10 nM had no effect. BQ-788, a selective ET(B) receptor antagonist, completely blocked stimulation of eNOS expression caused by 0.1 nM ET-1 (12 +/- 25 vs. 120 +/- 40% for ET-1 alone; P < 0.05; n = 5). BQ-123, a selective ET(A) receptor antagonist, did not affect the increase in eNOS caused by 0.1 nM ET-1. Sarafotoxin c (S6c; 0.1 microM), a selective ET(B) receptor agonist, increased eNOS expression by 77 +/- 30% (P < 0.05; n = 6). Wortmannin (0.01 microM), a PI3K inhibitor, completely blocked the stimulatory effect of 0.1 microM S6c (77 +/- 30 vs. -28 +/- 9%; P < 0.05; n = 6). To test whether the increase in eNOS expression heightens activity, we measured NO release in response to simultaneous treatment with l-arginine, ionomycin, and clonidine using a NO-sensitive electrode. NO release by control cells was 337 +/- 61 and 690 +/- 126 pA in ET-1-treated cells (P < 0.05; n = 5). Taken together, these data suggest that ET-1 stimulates THAL eNOS, activating ET(B) receptors and PI3K and thereby increasing NO production. PMID- 15113750 TI - Adriamycin impairs the contraction of mesangial cells through the inhibition of protein kinase C and intracellular calcium. AB - The effects of adriamycin on the contractile function of cultured mesangial cells were measured by the changes in planar surface area in response to treatment with agonists. Incubation of mesangial cells with adriamycin (0.2 microg/ml) for 24 h significantly decreased the contractile responses to the calcium channel activator BAY K 8644 (1 microM) and to the PKC activator PMA (1 microM). Intracellular calcium concentration ([Ca(2+)](i)), measured by changes in fura 2 levels in response to ATP (0.1 mM), was significantly inhibited in adriamycin treated mesangial cells compared with control cells. In the absence of extracellular calcium, treatment with ionomycin (0.1 mM) or thapsigargin (10 microM) resulted in a significantly smaller increase in [Ca(2+)](i) in adriamycin treated mesangial cells compared with control, suggesting an important role of the endoplasmic reticulum in the effects of adriamycin. Using PKC-specific antibodies, adriamycin significantly decreased the cytosolic and membranous fractions of PKC-alpha in mesangial cells to 75 +/- 6 and 70 +/- 12% of control, respectively. The PKC activity of mesangial cells was also significantly inhibited after incubation with adriamycin for 24 h. In conclusion, adriamycin induces hypocontractility of mesangial cells, which may mediate this effect by inhibiting PKC-alpha and [Ca(2+)](i). PMID- 15113751 TI - Inhibition of Na-K-ATPase in thick ascending limbs by NO depends on O2- and is diminished by a high-salt diet. AB - A high-salt diet enhances nitric oxide (NO)-induced inhibition of transport in the thick ascending limb (THAL). Long exposures to NO inhibit Na-K-ATPase in cultured cells. We hypothesized that NO inhibits THAL Na-K-ATPase after long exposures and a high-salt diet would augment this effect. Rats drank either tap water or 1% NaCl for 7-10 days. Na-K-ATPase activity was assessed by measuring ouabain-sensitive ATP hydrolysis by THAL suspensions. After 2 h, spermine NONOate (SPM; 5 microM) reduced Na-K-ATPase activity from 0.44 +/- 0.03 to 0.30 +/- 0.04 nmol P(i).microg protein(-1).min(-1) in THALs from rats on a normal diet (P < 0.03). Nitroglycerin also reduced Na-K-ATPase activity (P < 0.04). After 20 min, SPM had no effect (change -0.07 +/- 0.05 nmol P(i).microg protein(-1).min(-1)). When rats were fed high salt, SPM did not inhibit Na-K-ATPase after 120 min. To investigate whether ONOO(-) formed by NO reacting with O(2)(-) was involved, we measured O(2)(-) production. THALs from rats on normal and high salt produced 35.8 +/- 0.3 and 23.7 +/- 0.8 nmol O(2)(-).min(-1).mg protein(-1), respectively (P < 0.01). Because O(2)(-) production differed, we studied the effects of the O(2)(-) scavenger tempol. In the presence of 50 microM tempol, SPM did not inhibit Na-K-ATPase after 120 min (0.50 +/- 0.05 vs. 0.52 +/- 0.07 nmol P(i).microg protein(-1).min(-1)). Propyl gallate, another O(2)(-) scavenger, also prevented SPM-induced inhibition of Na-K-ATPase activity. SPM inhibited pump activity in tubules from rats on high salt when O(2)(-) levels were increased with xanthine oxidase and hypoxanthine. We concluded that NO inhibits Na-K-ATPase after long exposures when rats are on a normal diet and this inhibition depends on O(2)(-). NO donors do not inhibit Na-K-ATPase in THALs from rats on high salt due to decreased O(2)(-) production. PMID- 15113752 TI - The effect of high glucose and PPAR-gamma agonists on PPAR-gamma expression and function in HK-2 cells. AB - Peroxisome proliferator-activated receptor-gamma (PPAR-gamma) are ligand activated transcription factors that regulate cell growth, inflammation, lipid metabolism, and insulin sensitivity. PPAR-gamma in the human kidney has been described. However, the role of PPAR-gamma in proximal tubular cells with respect to cell growth and inflammation in diabetic nephropathy is largely unknown. We evaluated the effect of high (30 mM) D-glucose, thiazolidinedione pioglitazone (10 microM), and the selective PPAR-gamma agonist L-805645 (8 microM) on PPAR gamma expression, growth, and inflammatory parameters in the proximal tubular model of HK-2 cells. PPAR-gamma was present in HK-2 cells and upregulated with 30 mM D-glucose to 177 +/- 31.2% of control (P < 0.05). PPAR-gamma activation was induced by pioglitazone to a similar level to that observed by exposure to high glucose but maximally induced by the selective agonist L-805645. However, L 805645 reduced cell viability in both 5 and 30 mM d-glucose to 73.8 +/- 3.1 and 77.6 +/- 1.4% of control (both P < 0.0001). In parallel, thymidine incorporation was reduced with L-805645 in both 5 and 30 mM D-glucose to 33.3 +/- 3.4 and 37.9 +/- 2.2%, respectively (both P < 0.0001). Flow cytometry demonstrated increased apoptosis and G(1) phase arrest in association with an increase in p21(cip1/waf1) in cells exposed to L-805645. Exposure to 30 mM D-glucose did not significantly change AP-1 promoter activity (89.0 +/- 5.5% of control); however, the addition of L-805645 significantly reduced it to 62.2 +/- 2.7% of control (P < 0.0001). Thirty nanomolar D-glucose induced transforming growth factor-beta(1) to 137.7 +/ 16.9% of control (P < 0.05), and L-805645 was able to suppress this to 68.7 +/- 5.7% of control (P < 0.01 vs. d-glucose). Exposure to 30 mM D-glucose reduced monocyte chemoattractant protein 1 levels to 78.6 +/- 7.1% (P < 0.05) of control, with the reduction more marked in the presence of either pioglitazone (P < 0.01) or L-805645 (P < 0.01). In summary, high glucose upregulates PPAR-gamma and when significantly induced demonstrates anti-proliferative and anti-inflammatory effects. PMID- 15113753 TI - Interaction strategies of lesbian, gay, and bisexual healthcare practitioners in the clinical examination of patients: qualitative study. AB - OBJECTIVE: To explore how lesbian, gay, and bisexual healthcare practitioners manage their identity in the clinical examination of patients. DESIGN: Qualitative study using grounded theory. SETTING: Hospital and primary health care. PARTICIPANTS: 16 healthcare professionals who identified themselves as lesbian, gay, or bisexual, and are involved in the clinical examination of patients. RESULTS: Healthcare professionals engage in a complex interplay of identity management strategies to avoid homophobic abuse; as a signal of safety from homophobia and understanding for their lesbian, gay, and bisexual patients and as a desexualisation strategy principally for gay men and their women patients. Their training has not helped them deal with ethical and medicolegal anxieties. CONCLUSION: In the light of new legislation, published guidelines will help training and governing bodies understand and help ameliorate the added pressures on their lesbian, gay, and bisexual students and medical staff. PMID- 15113754 TI - LFA-1 signaling through p44/42 is coupled to perforin degranulation in CD56+CD8+ natural killer cells. AB - Leukocyte function antigen 1 (LFA-1) is essential for the formation of immune cell synapses and plays a role in the pathophysiology of various autoimmune diseases. We investigated the molecular details of LFA-1 activation during adhesion between cytotoxic cells and a target model leukemia cell. The cytolytic activity of a CD3-CD8+CD56+ natural killer (NK) subset was enhanced when LFA-1 was activated. In a comparison of LFA-1 ligands, intercellular adhesion molecule 2 (ICAM-2) and ICAM-3 promoted LFA-1-directed perforin release, whereas ICAM-1 had little effect. Ligand-induced LFA-1 clustering facilitated perforin release, demonstrating LFA-1 could regulate degranulation mechanisms. LFA-1 induced the activation of src family kinases, Vav1 and p44/42 mitogen-activated protein kinase (MAPK), in human CD56+ NK cells as evidenced by intracellular phospho epitope measurements that correlated with effector-target cell binding and perforin-granzyme A-mediated cytolytic activity. These results identify novel, specific functional consequence of LFA-1-mediated cytolytic activity in perforin containing human NK subsets. PMID- 15113755 TI - Natural killer T cells accelerate atherogenesis in mice. AB - We have investigated the potential role of CD1d-restricted natural killer T (NKT) cells in the development of atherosclerosis in mice. When fed an atherogenic diet (AD), NKT cell-deficient CD1d(-/-) mice had significantly smaller atherosclerotic lesions than AD-fed C57BL/6 (wild-type [WT]) mice. A significant reduction in atherosclerotic lesions was also demonstrated in AD-fed, low-density lipoprotein receptor-deficient (Ldlr(-/-)) mice reconstituted with CD1d(-/-) bone marrow cells compared with the lesions observed in Ldlr(-/-)mice reconstituted with WT marrow cells. In addition, repeated injections of alpha-GalCer or the related glycolipid OCH to apolipoprotein E knockout (apoE(-/-)) mice during the early phase of atherosclerosis significantly enlarged the lesion areas compared with mice injected with vehicle control. However, administering alpha-GalCer to apoE( /-) mice with established lesions did not significantly increase the lesion area but considerably decreased the collagen content. Atherosclerosis development in either AD-fed WT or apoE(-/-) mice was associated with the presence of Valpha14Jalpha18 transcripts in the atherosclerotic arterial walls, indicating that NKT cells were recruited to these lesions. Thioglycolate-elicited macrophages pulsed with oxidized low-density lipoproteins expressed enhanced CD1d levels and induced NKT cells to produce interferon-gamma, a potentially proatherogenic T-helper 1 (TH1) cytokine. Collectively, we conclude that NKT cells are proatherogenic in mice. PMID- 15113756 TI - The etiology of severe anemia in a village and a periurban area in Mali. AB - Severe anemia is one of the major complications of malaria in Africa. We studied 2 populations, one in a village and the second in a periurban area in Mali, to understand the preventable factors in the disease. The 2 correlates of disease were parasitemia above 100 000 parasitized red blood cells per microliter (0.1 x 10(12)/L) and a low baseline hemoglobin level. All cases of moderate to severe anemia occurred in children under 3.2 years of age. Raising the baseline hemoglobin level and lowering peak parasitemia in infants and young children may reduce the incidence of severe anemia resulting from malarial infection. PMID- 15113757 TI - Immunoregulation of dendritic cells by IL-10 is mediated through suppression of the PI3K/Akt pathway and of IkappaB kinase activity. AB - Interleukin-10 (IL-10) has potent immunoregulatory effects on the maturation and the antigen-presenting cell (APC) function of dendritic cells (DCs). The molecular basis underlying these effects in DCs, however, is ill defined. It is well established that the transcription factor NF-kappaB is a key regulator of DC development, maturation, and APC function. This study was initiated to determine the effects of IL-10 on the NF-kappaB signaling pathway in immature DCs. IL-10 pretreatment of myeloid DCs cultured from bone marrow resulted in reduced DNA binding and nuclear translocation of NF-kappaB after anti-CD40 antibody or lipopolysaccharide (LPS) stimulation. Furthermore, inhibited NF-kappaB activation was characterized by reduced degradation, phosphorylation, or both of IkappaBalpha and IkappaBepsilon but not IkappaBbeta and by reduced phosphorylation of Ser536, located in the trans-activation domain of p65. Notably, IL-10-mediated inhibition of NF-kappaB coincided with suppressed IkappaB kinase (IKK) activity in vitro. Furthermore, IL-10 blocked inducible Akt phosphorylation, and inhibitors of phosphatidylinositol 3-kinase (PI3K) effectively suppressed the activation of Akt, IKK, and NF-kappaB. These findings demonstrate that IL-10 targets IKK activation in immature DCs and that suppressing the PI3K pathway in part mediates blockade of the pathway. PMID- 15113758 TI - Safety and efficacy of denileukin diftitox in patients with steroid-refractory acute graft-versus-host disease after allogeneic hematopoietic stem cell transplantation. AB - Denileukin diftitox (Ontak), a recombinant protein composed of human interleukin 2 (IL-2) fused to diphtheria toxin, has selective cytotoxicity against activated lymphocytes expressing the high-affinity IL-2 receptor. We conducted a phase 1 study of denileukin diftitox in 30 patients with steroid refractory acute graft versus-host disease (GVHD). Seven patients received 9 microg/kg intravenously on days 1 and 15; 18 received 9 microg/kg intravenously on days 1, 3, 5, 15, 17, and 19; and 5 received 9 microg/kg intravenously on days 1 to 5 and 15 to 19. Hepatic transaminase elevation was the dose-limiting toxicity (DLT), and dose level 2 was the maximum tolerated dose (MTD). Overall, 71% of patients responded with complete resolution (12 of 24; 50%) or partial resolution (5 of 24; 21%) of GVHD. Eight of 24 patients (33%) are alive at 6.3 to 24.6 months (median, 7.2 months). Denileukin diftitox is tolerable and has promising activity in steroid-refractory acute GVHD. PMID- 15113759 TI - Comparing morbidity and mortality of HLA-matched unrelated donor hematopoietic cell transplantation after nonmyeloablative and myeloablative conditioning: influence of pretransplantation comorbidities. AB - We have carried out HLA-matched unrelated donor hematopoietic cell transplantation (HCT) after nonmyeloablative conditioning in patients with hematologic malignancies who were ineligible for conventional transplantations because of age, comorbidities, or both. The nonmyeloablative regimen consisted of 90 mg/m2 fludarabine and 2 Gy total body irradiation given before and mycophenolate mofetil and cyclosporine given after HCT. This report compares, retrospectively, morbidity and mortality among 60 consecutive patients given nonmyeloablative conditioning (nonablative patients) to those among 74 concurrent and consecutive patients given myeloablative conditioning (ablative patients) before unrelated HCT. The Charlson Comorbidity Index was used to assess pretransplantation comorbidities. Even though nonablative patients had significantly higher pretransplantation comorbidity scores, were older, and had more often failed preceding ablative transplantations and cytotoxic therapies, they experienced fewer grades III to IV toxicities than ablative patients. Further, the incidence of grades III to IV acute graft-versus-host disease (GVHD) was significantly lower in nonablative patients. Both patient groups had comparable 1-year probabilities of chronic GVHD. The 1-year nonrelapse mortality rate was 20% in nonablative patients compared to 32% in ablative patients (hazard ratio=1.4). After adjustment for pretransplantation differences between the 2 patient groups, the hazard ratio was 3.0 (P=.04). Multivariate analyses showed higher pretransplantation comorbidity scores to result in increased toxicity and mortality. PMID- 15113760 TI - Tuning the volume of the immune response: strength and persistence of stimulation determine migration and cytokine secretion of dendritic cells. AB - Migration to lymph nodes and secretion of cytokines are critical functions of mature dendritic cells (DCs); however, these 2 functions are not necessarily linked. This is the first report showing that quantitative differences in identical signaling pathways determine DC migration and cytokine secretion. Using different polymerized forms of CD40 ligand, we demonstrate that the strength and persistence of CD40 signaling can induce either function. Induction of monocyte derived DC (MoDC) migration required a weak and transient CD40 signal, whereas strong and persistent CD40 signaling blocked migration and biased toward cytokine secretion. In contrast to MoDCs, CD40 activation of CD1c+ peripheral blood DCs (PBDCs) induced a nonpersistent, intracellular signaling profile resulting in migratory-type DCs unable to secrete interleukin-12p70 (IL-12p70). Extracellular signal-regulated kinase 1/2 (ERK1/2) and p38K activation synergistically mediated cytokine secretion, whereas migration was enhanced by p38K activation but reduced by persistent ERK1/2 activity. This model of signal strength and persistence also applied when stimulating DCs with intact microbes. Thus, a novel concept emerges in which the type of immune response induced by DCs is tuned by the strength and persistence of DC activating signals. PMID- 15113761 TI - Continuous in vivo infusion of interferon-gamma (IFN-gamma) preferentially reduces myeloid progenitor numbers and enhances engraftment of syngeneic wild type cells in Fancc-/- mice. AB - Fanconi anemia (FA) is characterized by bone marrow (BM) failure and cancer susceptibility. Identification of the cDNAs of many FA complementation types allows the potential of using gene transfer technology to introduce functional cDNAs as transgenes into autologous stem cells and provide a cure for the BM failure in FA patients. Previous studies in FA murine models and in a phase 1 clinical trial suggest that myelopreparation is required for significant engraftment of exogenous, genetically corrected stem cells. Since myeloid progenitors from Fancc-/- mice and human Fanconi anemia group C protein (FANCC) patients have increased apoptosis in response to interferon gamma (IFN-gamma) in vitro, we hypothesized that IFN-gamma may be useful as a nongenotoxic, myelopreparative conditioning agent. To test this hypothesis, IFN-gamma was administered as a continuous infusion to Fancc-/- and wild-type (WT) mice for 1 week. Primitive and mature myeloid lineages were preferentially reduced in IFN gamma-treated Fancc-/- mice. Further, IFN-gamma conditioning of Fancc-/- recipients was sufficient as a myelopreparative regimen to allow consistent engraftment of isogenic WT repopulating stem cells. Collectively, these data demonstrate that Fancc-/- hematopoietic cell populations have increased hypersensitivity to IFN-gamma in vivo and that IFN-gamma conditioning may be useful as a nongenotoxic strategy for myelopreparation in this disorder. PMID- 15113762 TI - Death and its diagnosis by doctors. PMID- 15113763 TI - Prevention of cardiac complications of non-cardiac surgery: stenosis and thrombosis. PMID- 15113764 TI - Difficulty in brainstem death testing in the presence of high spinal cord injury. AB - In the UK, when the standard brain death criteria are met, further investigations are not necessary. Confirmatory tests can be useful, however, when it is not possible to carry out all of the brainstem tests. We report the case of a patient with multiple trauma and a high spinal cord injury who was apnoeic. Confirmatory tests (EEG, brainstem, auditory evoked potential) were essential in supporting the diagnosis of brainstem death to allow withdrawal of artificial ventilation, as organ donation was being considered. PMID- 15113765 TI - Obstetric epidural and chronic adhesive arachnoiditis. PMID- 15113766 TI - Post-dural puncture headache: pathogenesis, prevention and treatment. PMID- 15113767 TI - Anaesthesia for spinal surgery in adults. PMID- 15113768 TI - Bispectral analysis gives us more information than power spectral-based analysis. PMID- 15113769 TI - Different actions of sevoflurane and propofol on central nicotinic receptors may explain differences in hypnotic antagonism by cholinesterase inhibitors. PMID- 15113770 TI - Correct nomenclature of superficial cervical plexus blocks. PMID- 15113771 TI - Perioperative fluid optimization. PMID- 15113772 TI - PFA-100 and regional analgesia in a parturient after ibuprofen overdose. PMID- 15113773 TI - The CobraPLA in 110 anaesthetized and paralysed patients: what size to choose? PMID- 15113774 TI - Expression of C-type lectin receptors by subsets of dendritic cells in human skin. AB - C-type lectins are cell surface receptors that recognize carbohydrate structures which are often part of microbial pathogens. Several of these molecules are expressed on dendritic cells and are involved in antigen uptake. Expression of C type lectins on dendritic cells of the human skin, i.e. Langerhans cells of the epidermis and dermal dendritic cells, has been incompletely studied to date. We therefore investigated C-type lectins in situ and on dendritic cells obtained by migration from skin explants by immunofluorescence and flow cytometry. Emphasis was laid on expression patterns of DEC-205/CD205 and BDCA-2, a marker for plasmacytoid dendritic cells. Langerhans cells in situ expressed low levels of DEC-205. Expression was upregulated upon maturation in skin explant organ culture. Most dermal dendritic cells were found to be positive for DEC-205 and DC SIGN/CD209. Few BDCA-2-expressing cells were found in most skin samples. They were located in small groups in the dermis close beneath the basement membrane. The vast majority of all types of dendritic cells in normal human skin was of immature phenotype, i.e. did not express DC-LAMP/CD208. It is concluded that normal appearing human skin harbors different subsets of dendritic cells including few scattered BDCA-2-expressing cells, presumably plasmacytoid dendritic cells, expressing variable sets of C-type lectin receptors. This may critically contribute to the capacity of the skin immune system to flexibly respond to the world of microbial pathogens. PMID- 15113775 TI - Diabetes in the Department of Veterans Affairs. PMID- 15113776 TI - Diabetes in nonveterans, veterans, and veterans receiving Department of Veterans Affairs health care. AB - OBJECTIVE: To compare behavioral risk factors and health and disease characteristics among three groups of adults with diabetes: nonveterans, veterans not receiving Department of Veterans Affairs (VA) health care, and veterans using VA services. RESEARCH DESIGN AND METHODS: Two data sources were used to describe the veteran population. First, the 2000 Behavioral Risk Factor Surveillance System (BRFSS) characterized the U.S. adult population by preventive health practices and risk behaviors linked to chronic and preventable diseases. New to the 2000 survey were questions on veteran status, which were administered in all states. Second, VA administrative and veterans benefits data were analyzed to describe comorbidity, education services, and veterans benefits. RESULTS: The estimated prevalence of diabetes in male veterans receiving VA care was 16%. Male veterans with diabetes using VA care were more likely to be nonwhite, not employed, have lower income, lower health status, and more activity limitations than male veterans not using these services. Computerized records indicate VA users with diabetes also had high concurrent comorbidity. Frequency of VA diabetes and preventive care services, as measured by selected quality indicators, was equivalent to or higher than the levels reported by veterans not receiving VA care and nonveterans. In addition to health care, nearly one-fourth of veterans with diabetes also received monthly awards for compensation and pension. CONCLUSIONS: Males receiving VA care with self-reported diabetes indicated receiving preventive care services at equivalent or higher levels than their counterparts receiving care outside the VA and nonveterans. PMID- 15113777 TI - Who has diabetes? Best estimates of diabetes prevalence in the Department of Veterans Affairs based on computerized patient data. AB - OBJECTIVE: To optimize methods for identifying patients with diabetes based on computerized records and to obtain best estimates of diabetes prevalence in Department of Veterans Affairs (VA) patients. RESEARCH DESIGN AND METHODS: The VA Diabetes Epidemiology Cohort (DEpiC) is a linked national database of all VA patients since 1998 with data from VA medical visits, Medicare claims, pharmacy and laboratory records, and patient surveys. Using DEpiC, we examined concordance of diabetes indicators, including ICD-9-CM codes (250.xx), prescription drug treatment, HbA(1c) tests, and patient self-report. We determined the optimal criterion for identifying diabetes and used it in estimating diabetes prevalence in the VA. RESULTS: The best criterion was a prescription for a diabetes medication in the current year and/or 2+ diabetes codes from inpatient and/or outpatient visits (VA and Medicare) over a 24-month period. This definition had high sensitivity (93%) and specificity (98%) against patient self-report, and reasonable rates of HbA(1c) testing (75%). HbA(1c) testing alone added few additional cases, and a single diagnostic code added many patients, but without confirmation (reduced specificity). However, including codes from Medicare was critical. Applying this definition for 1998-2000, we identified an average of 500,000 VA patients with diabetes per year. We also estimated high and increasing diabetes prevalence rates of 16.7% in FY1998, 18.6% in FY1999, and 19.6% in FY2000 and an incidence estimated to be approximately 2% per year. CONCLUSIONS: Development and evaluation of methodology for analyzing computerized patient data can improve the identification of patients with diabetes. The increasing high prevalence of diabetes in VA patients will present challenges for clinicians and health system management. PMID- 15113778 TI - Data resources in the Department of Veterans Affairs. AB - OBJECTIVE: To provide an overview of databases that are maintained by the Department of Veterans Affairs (VA) and are of relevance to investigators involved in epidemiologic, clinical, and health services research. RESEARCH DESIGN AND METHODS: We reviewed both national and local VA databases and identified their strengths and limitations. We also referenced specific studies that have assessed the validity and reliability of VA databases. RESULTS: There are numerous national databases housed at the Austin Automation Center in Austin, Texas. These include the Patient Treatment File (hospital abstracts), the Outpatient Care File, the Beneficiary Identification Record Locator System death file for assessing vital status, and the Decision Support System, which provides integrated clinical and financial information for managerial decision making. The major limitation of these databases is that clinical detail below the level of ICD-9-CM diagnosis and procedure codes is not uniformly available nationally. These databases offer an excellent opportunity to monitor the health of veterans over time because they track all inpatient and outpatient utilization in the VA. However, at the local or medical center level, the Veterans Health Information and Systems and Technology Architecture contains extensive clinical information, but has fewer patients, varies in format across medical centers, and poses difficulties with data extraction for statistical analysis. CONCLUSIONS: Both local and national VA databases are valuable resources for investigators who have interests in a wide array of research topics, including diabetes. The potential for investigating important scientific questions with VA databases becomes greater as communications and database management technologies improve. PMID- 15113779 TI - The burden of diabetes-associated cardiovascular hospitalizations in Veterans Administration (VA) and non-VA medical facilities. AB - OBJECTIVE: This study examines the relative burden of cardiovascular disease in diabetic and nondiabetic admissions to Veterans Administration (VA) and non-VA hospitals. RESEARCH DESIGN AND METHODS: Calendar year 1997 hospitalization data were collected from the VA Patient Treatment File and the National Hospital Discharge Survey, National Center for Health Statistics. Discharge diagnoses based on ICD-9 codes were used to classify cardiovascular events and procedures and diabetes. Proportionate hospitalization ratios (PHRs) were calculated based on the number of cardiovascular hospitalizations from among all hospitalizations. RESULTS: We identified 119,653 VA hospitalizations (19.1%) and 3,765,696 non-VA hospitalizations (16.8%) that coded diabetes among the discharge diagnoses. Cardiovascular hospitalizations, primarily coronary in origin, accounted for nearly 50% of all hospitalizations of persons with diabetes within VA and non-VA medical care systems. Coronary events and procedures (PHR = 1.85 and 1.68) and, to a lesser extent, cerebrovascular events and procedures (PHR = 1.55 and 1.33) were more common in VA hospitalizations where diabetes was listed as a comorbidity than in VA hospitalizations where diabetes was not listed. The burden of coronary hospitalizations was larger in VA than non-VA facilities for men (PHR = 1.82 vs. 1.66) and smaller in VA than non-VA facilities for women (PHR = 3.11 vs. 3.44) in age and race-standardized analyses. The burden of coronary hospitalization was higher for whites and blacks in VA facilities (PHR = 1.83 and 2.01) when compared with non-VA facilities (PHR = 1.68 and 1.84). CONCLUSIONS: The burden of cardiovascular hospitalizations is not equally borne across hospitalizations with and without diabetes and across VA and non-VA facilities. PMID- 15113780 TI - Cardiovascular risk factor control among veterans with diabetes: the ambulatory care quality improvement project. AB - OBJECTIVE: To describe the extent to which hyperglycemia, hypertension, and dyslipidemia are currently detected, treated, and controlled in U.S. veterans with diabetes with and without ischemic heart disease (IHD). RESEARCH DESIGN AND METHODS: A cohort of 3,769 veterans who self-reported diabetes and who received all health care from the Veterans Administration (VA) medical centers were selected from subjects enrolled in the Ambulatory Care Quality Improvement Project, a randomized health services intervention at seven VA primary care clinics. IHD was defined by a self-reported history of myocardial ischemia, infarction, or revascularization. Mean values of HbA(1c), blood pressure, and cholesterol subfractions were collected from computerized laboratory databases. Medication data were collected from computerized pharmacy databases. RESULTS: Mean HbA(1c) and optimal control (HbA(1c) <7%) did not differ for those without and with IHD: 8.1 vs. 8.0%, and 26 vs. 24%, respectively. Veterans with IHD were more likely to have hypertension (73 vs. 64%), to be treated (88 vs. 78%), and to have optimal blood pressure control (19 vs. 10%) compared with veterans without IHD (all P values <0.01). Veterans with IHD were more likely to have dyslipidemia (81 vs. 53%), were equally likely to be treated (54 vs. 50%), and were more likely to have optimal LDL levels (30 vs. 16%) compared with veterans without IHD, all P values <0.01. CONCLUSIONS: Optimal cardiovascular risk factor control was the exception in this cohort of diabetic veterans attending primary care clinics. More aggressive management of cardiovascular risk factors in veterans with diabetes may be warranted, especially among those without prevalent IHD. PMID- 15113781 TI - The epidemiology of lower-extremity disease in veterans with diabetes. AB - OBJECTIVE: To describe the epidemiology of lower-extremity complications of diabetes in veterans who are users of the Department of Veterans Affairs (VA). RESEARCH DESIGN AND METHODS: Hospital discharge records for care provided in all VA hospitals in 1998 were obtained. All hospitalizations for lower-extremity ulceration, peripheral vascular procedures, and amputation were analyzed using frequency tables. A diabetes denominator was defined as a veteran with at least three ambulatory care visits with at least one diabetes diagnosis code. Age specific and total age-adjusted rates of discharge with ulceration, vascular procedures, and amputation were calculated. RESULTS: Veterans with diabetes comprised over half of all hospitalizations for lower-extremity ulceration, one third of all hospitalizations for peripheral vascular procedures, and two-thirds of all hospitalizations for amputation. The age-specific discharge rate per 1,000 diabetic persons for age 0-64 years, 65-74 years, and 75 years and older for ulceration were 28.4, 31.0, and 37.9; for vascular procedures, the rates were 3.5, 4.4, and 4.4; and for amputation, the rates were 7.3, 9.0, and 10.0, respectively. CONCLUSIONS: Veterans with diabetes comprise a significant proportion of hospitalizations for lower-extremity ulceration, peripheral vascular bypass, and amputation. Age-specific rates of diabetic amputation in veterans are lower than U.S. rates. PMID- 15113782 TI - Diabetes and renal disease in veterans. AB - OBJECTIVE: To describe the frequency and sequelae of diabetic renal disease in veterans who receive health care from the Veterans Administration (VA). RESEARCH DESIGN AND METHODS: Veterans with a diagnosis of diabetes, diabetic nephropathy, other kidney diseases, and end-stage renal disease (ESRD) were identified by diagnosis codes from national VA databases for FY1998. Data were obtained and analyzed for prevalence of renal disease, comorbid conditions, and 1-year mortality. RESULTS: A total of 44,671 (10.7%) of the 415,910 veterans with diabetes had a concomitant diagnosis of any renal disease. The average age was 67 years; 98% were male and 60% were white. The prevalence of diabetic nephropathy was 6.0% (n = 25,263). ESRD secondary to diabetes was present in 4.2% (17,636) of subjects. The age-standardized prevalence of diabetes and any renal disease was 72.6/1,000 persons and differed by race (white 76.1/1,000, black 103.4/1,000 persons). Diabetes-associated ESRD prevalence was higher among black versus white veterans and male versus female veterans. One-year age-standardized mortality was 10.7%. CONCLUSIONS: Nephropathy is prevalent in veterans with diabetes. Greater mortality is observed among those with renal disease compared with those without renal disease. Additional surveillance is needed to identify persons likely to progress to diabetic nephropathy and to plan for appropriate and timely health care for these individuals. PMID- 15113783 TI - Eye disease in veterans with diabetes. AB - OBJECTIVE: To describe the screening, prevalence, and management of eye disease in veterans cared for by the Veterans Affairs (VA) System. RESEARCH DESIGN AND METHODS: Eye examinations, treatments, and diseases were identified in veterans with diabetes who received care in inpatient and outpatient VA settings during FY1998. Analysis was conducted to characterize the patient population and screened population and to compare them to the total VA patient population with diabetes. Logistic regression was performed to predict eye screening. RESULTS: Overall 48% of veterans with diabetes had an eye examination in FY1998. One-third of all veterans had an eye condition and 8.6% of veterans had ophthalmic manifestations of diabetes. In addition 11% reported glaucoma and 17.8% reported cataract surgery. Approximately 11,500 (2.7%) veterans were blind. CONCLUSIONS: Visual impairment is a common complication of diabetes. Half of this population of veterans had a visual examination recorded in VA. The VA is testing a more targeted screening and treatment approach for diabetic eye disease in order to prevent the serious eye complications. PMID- 15113784 TI - Quality improvement initiatives: issues in moving from diabetes guidelines to policy. AB - OBJECTIVE: To outline the principles that direct the Veterans Affairs (VA) Quality Enhancement Research Initiative (QUERI) dedicated to diabetes quality improvement (QUERI-DM). RESEARCH DESIGN AND METHODS: We discuss the VA initiatives aimed at improving diabetes care for veterans as well as general issues that should be considered in quality improvement initiatives. We specifically describe some of the epidemiological, statistical, and organizational issues that have guided our quality improvement (QI) programs. RESULTS: The five principles that have guided the QUERI-DM process are: 1) treating clinical guidelines and goals distinct from quality standards and quality improvement priorities; 2) targeting high-risk patients and high-impact quality issues; 3) profiling processes over outcomes; 4) targeting processes that will improve patient outcomes; and 5) paying attention to the loci of practice variation. CONCLUSIONS: The authors recommend that all five principles be considered when moving from practice guidelines to performance measures and QI initiatives. Targeting high-priority problems and high-risk groups can greatly improve the effectiveness and efficiency of QI interventions. PMID- 15113785 TI - Diabetes quality improvement in Department of Veterans Affairs Ambulatory Care Clinics: a group-randomized clinical trial. AB - OBJECTIVE: To conduct a group-randomized clinical trial to determine whether regular feedback to primary care providers of synthesized information on patients' health, function, and satisfaction would demonstrate improved outcomes for their patients with diabetes. RESEARCH DESIGN AND METHODS: Patients in General Internal Medicine Clinics Department of Veterans Affairs (VA) Medical Centers were randomized into seven intervention or control firms. Patient self reported information was collected by mail on general health, diabetes, and up to five other chronic conditions. Patients with diabetes received the Seattle Diabetes Questionnaire, the 36-item Medical Outcomes Study short form (SF-36), and a validated patient satisfaction questionnaire at regular intervals. Data from self-report, clinical, pharmacy, and laboratory sources were synthesized into patient-specific feedback reports that intervention providers received before patients' visits. RESULTS: The timely delivery to primary care providers of state-of-the-art patient-feedback reports that identified patient issues and areas for improvement did not result in significant improvements in patient outcomes between the intervention and control firms. CONCLUSIONS: Outcomes in diabetic patients whose providers received synthesized patient data before visits were no better than in those receiving care from control firms. Future studies may benefit from substantial involvement in patients discussing, problem solving, and goal setting in addition to use of timely synthesized patient data. PMID- 15113786 TI - Diabetes-related utilization and costs for inpatient and outpatient services in the Veterans Administration. AB - OBJECTIVE: -The purpose of this study was to calculate the total number of inpatient hospitalizations, outpatient clinic visits, and total direct health care costs associated with veterans with diabetes receiving care in Veterans Administration (VA) facilities. RESEARCH DESIGN AND METHODS-The number of inpatient hospitalizations is tracked for years 1994-1998, and outpatient clinic visits are tracked for years 1997 and 1998. Trends in utilization across different age and racial groups, as well as total direct inpatient and outpatient costs for 1998, are presented. RESULTS: -Between 1994 and 1998, hospitalization rates decreased from 1.68 to 1.61. The average number of outpatient visits was 4.5 in 1997 and 4.6 in 1998. VA incurred $214.8 million in outpatient expenditures and $1.45 billion in inpatient expenditures for veterans with diabetes receiving VA care. CONCLUSIONS: -Health care delivery systems and payors track the cost and utilization of services by specific patient groups to support disease management, quality improvement, external reporting, and cost containment. Tracking the utilization and cost of diabetes care is necessary to understand the financial impact of diabetes on health care systems and the overall burden of diabetes on individuals. PMID- 15113787 TI - Pharmacy costs and glycemic control in the Department of Veterans Affairs. AB - OBJECTIVE: To determine pharmacy costs for glycemic treatment and its relationship to glycemic control in the Department of Veterans Affairs (VA) between 1994 and 2000. RESEARCH DESIGN AND METHODS: Patients with diabetes in the VA in FY1994, FY1996, FY1998, and FY2000 were identified using an ambulatory care pharmacy-derived database. Total drug acquisition costs, as well as expenditures for insulin, oral glycemic control agents, and self-blood glucose monitoring strips, were determined for these veterans. HbA(1c) levels for the corresponding time periods were also obtained. Pharmacy costs (medications and monitoring) were examined by glycemic control treatment type. RESULTS: In FY2000, 18% (n = 535,016) of all VA pharmacy patients were identified as having diabetes, and they received 30% of all pharmacy prescriptions. Overall, 23% of pharmacy expenditures for these patients were related to glycemic control medications and monitoring supplies. Annual pharmacy costs increased from FY1994 to FY2000. The greatest change was the higher expenditure for monitoring supplies through FY1998, which then decreased in FY2000. Increased pharmacy costs were associated with improved glycemic control. In FY2000, the mean last HbA(1c) level (n = 446,384) fell to 7.6% from 7.8% in FY1998 (n = 204,136) and 8.4% in 1996 (n = 53,348). CONCLUSIONS: Diabetes was associated with high pharmacy costs. Increasing medication expenditures were associated with improved HbA(1c) levels at the aggregated national level. Policies concerning dispensing monitoring supplies and several diabetes quality improvement projects were initiated during this interval. Future challenges include initiatives to further optimize care while controlling costs. PMID- 15113788 TI - Development of evidence-based clinical practice guidelines for diabetes: the Department of Veterans Affairs/Department of Defense guidelines initiative. AB - OBJECTIVE: To describe the Veterans Affairs (VA)/Department of Defense (DoD) Clinical Practice Guidelines for diabetes and contrast selected recommendations with those of the American Diabetes Association (ADA). RESEARCH DESIGN AND METHODS: We summarize the general structure of the VA/DoD Guidelines and describe the rationale for recommendations issued in 2003 for glycemic control, management of hypertension, and retinopathy screening. We compare the synthesis of evidence and resulting recommendations for these content areas with the 2004 American Diabetes Association Clinical Practice Recommendations. RESULTS: The VA/DoD Guidelines and the ADA Clinical Practice Recommendations reported similar strength of evidence findings by content area, but clinical recommendations varied. The VA/DoD Guidelines and practice recommendations emphasize the use of data on absolute risk reduction from available published randomized clinical trials rather than relative risk reduction from observational analyses. The VA/DoD Guidelines employ an algorithm-based methodology to guide clinicians through a risk-stratified approach to managing individual patients rather than promoting a single standard for most or all patients without explicit consideration of competing comorbidities. CONCLUSIONS: The VA/DoD Guidelines are intended to guide diabetes care by providing Internet-ready, evidence-based annotations in algorithmic form to help clinicians set and revise individual treatment goals for their patients. PMID- 15113789 TI - Diabetes process and outcome measures in the Department of Veterans Affairs. AB - OBJECTIVE: To evaluate performance of process and outcome measures in the care of patients with diabetes seen in Department of Veterans Affairs (VA) facilities. RESEARCH DESIGN AND METHODS: Retrospective audits of records and databases were conducted on randomly selected patients with diabetes over 5 years (1995 [baseline] and 1997-2000) in 22 VA networks. Performance on diabetes-specific and preventive processes was measured. RESULTS: Nationally, significant improvements over time were observed for all measures (P < 0.001). For example, the percentage of patients receiving a dilated retinal examination rose from 44% in 1995 to 67% in 2000. The percentage of patients who received a urinary protein test rose from 23% in 1997 to 54% in 2000. Those who received influenza vaccination rose from 34% in 1995 to 78% in 2000. However, there was significant regional variation among all measures. CONCLUSIONS: Adherence to diabetes-specific and preventive care measures in the VA improved from 1997 to 2000 compared with a 1995 baseline. The improvement occurred in a setting of the provision of guidelines, the contractual setting of specific targets, and the timely feedback of results to medical center and network directors. Future studies are needed to determine whether adherence to these measures will decrease the rates of complications in VA patients with diabetes. PMID- 15113790 TI - Diabetes research in the Department of Veterans Affairs. AB - OBJECTIVE: To provide an overview of the Department of Veterans Affairs (VA) research activities, highlighting diabetes-related research. RESEARCH DESIGN AND METHODS: Diabetes is an important component of the VA research portfolio. All four VA research services support aspects of diabetes research. VA diabetes research projects and funding were examined from 1998 to 2003. RESULTS: VA scientists are conducting research on diabetes genetics, etiology, diagnosis, therapy, epidemiology, health services, and rehabilitation. VA research funding is available to answer important veteran-relevant questions through peer review, Center of Excellence activities, and multisite trial mechanisms. Many VA scientists also receive research support from nonfederal sources, including private corporations and nonprofit foundations. The VA Office of Research and Development actively supports training the next generation of researchers through their career development awards and the VA health profession training programs. CONCLUSIONS: The VA's diabetes research portfolio is extensive and includes many investigators, trainees, and fellows. There is substantial leveraging of VA diabetes research with support from other federal and nonfederal funding agencies, foundations, and private corporations. VA diabetes research findings benefit the global diabetes care community. PMID- 15113791 TI - A piece of my mind. The scar. PMID- 15113792 TI - Donald M. Berwick, MD, MPP: advocate for evidence-based health system reform. Interview by Brian Vastag. PMID- 15113793 TI - Vaccine rumors, funding shortfall threaten to derail global polio eradication efforts. PMID- 15113794 TI - Similar drug names a risky prescription. PMID- 15113795 TI - Journals' freedom to publish affirmed. PMID- 15113804 TI - Supplemental oxygen and risk of surgical site infection. PMID- 15113805 TI - Supplemental oxygen and risk of surgical site infection. PMID- 15113806 TI - Supplemental oxygen and risk of surgical site infection. PMID- 15113807 TI - Supplemental oxygen and risk of surgical site infection. PMID- 15113808 TI - Supplemental oxygen and risk of surgical site infection. PMID- 15113809 TI - Supplemental oxygen and risk of surgical site infection. PMID- 15113810 TI - Ranolazine as add-on therapy for patients with severe chronic angina. PMID- 15113811 TI - Ranolazine as add-on therapy for patients with severe chronic angina. PMID- 15113812 TI - Community-acquired methicillin-resistant Staphylococcus aureus. PMID- 15113813 TI - Combined antiretroviral therapy during pregnancy and risk of congenital malformations. PMID- 15113814 TI - Effects of tolvaptan, a vasopressin antagonist, in patients hospitalized with worsening heart failure: a randomized controlled trial. AB - CONTEXT: Nearly 1 million hospitalizations for chronic heart failure occur yearly in the United States, with most related to worsening systemic congestion. Diuretic use, the mainstay therapy for congestion, is associated with electrolyte abnormalities and worsening renal function. In contrast to diuretics, the vasopressin antagonist tolvaptan may increase net volume loss in heart failure without adversely affecting electrolytes and renal function. OBJECTIVE: To evaluate the short- and intermediate-term effects of tolvaptan in patients hospitalized with heart failure. DESIGN, SETTING, AND PARTICIPANTS: Randomized, double-blind, placebo-controlled, parallel-group, dose-ranging, phase 2 trial conducted at 45 centers in the United States and Argentina and enrolling 319 patients with left ventricular ejection fraction of less than 40% and hospitalized for heart failure with persistent signs and symptoms of systemic congestion despite standard therapy. INTERVENTION: After admission, patients were randomized to receive 30, 60, or 90 mg/d of oral tolvaptan or placebo in addition to standard therapy, including diuretics. The study drug was continued for up to 60 days. MAIN OUTCOME MEASURES: In-hospital outcome was change in body weight at 24 hours after randomization; outpatient outcome was worsening heart failure (defined as death, hospitalization, or unscheduled visits for heart failure) at 60 days after randomization. RESULTS: Median (interquartile range) body weight at 24 hours after randomization decreased by -1.80 (-3.85 to -0.50), -2.10 (-3.10 to -0.85), -2.05 (-2.80 to -0.60), and -0.60 (-1.60 to 0.00) kg in the groups receiving tolvaptan 30, 60, and 90 mg/d, and placebo, respectively (P< or =.008 for all tolvaptan groups vs placebo). The decrease in body weight with tolvaptan was not associated with changes in heart rate or blood pressure, nor did it result in hypokalemia or worsening renal function. There were no differences in worsening heart failure at 60 days between the tolvaptan and placebo groups (P =.88 for trend). In post hoc analysis, 60-day mortality was lower in tolvaptan treated patients with renal dysfunction or severe systemic congestion. CONCLUSION: Tolvaptan administered in addition to standard therapy may hold promise for management of systemic congestion in patients hospitalized for heart failure. PMID- 15113815 TI - HER-2 testing in breast cancer using parallel tissue-based methods. AB - CONTEXT: Testing for HER-2 oncogene in breast cancer has increased because of its role as a prognostic and predictive factor. Some advocate gene testing by fluorescence in situ hybridization (FISH) vs protein testing by immunohistochemistry as the method which most accurately evaluates and predicts response to the anti-HER-2 antibody, trastuzumab. However, critical examination of FISH on a screening basis has yet to be performed. OBJECTIVES: To determine the correlation between FISH and immunohistochemistry results by determining HER 2/neu gene status on tumor sections with indeterminate immunohistochemistry results (2+ score), confirm gene amplification on tumor sections with positive results (3+ score), and verify gene status on tumor sections with negative results (0 or 1+ score). DESIGN, SETTING, AND PATIENTS: A quality control and quality assurance program for HER-2 testing by FISH, which used tumor specimens from 2963 patients (median age, 56 years) with breast cancer received from 135 hospitals and cancer centers in 29 states, was performed at a reference laboratory from January 1, 1999, to May 15, 2003. Every specimen evaluated by FISH was parallel tested with immunohistochemistry tests. MAIN OUTCOME MEASURES: With FISH as the presumed standard testing method, the positive and negative predictive values and sensitivity and specificity of immunohistochemistry were calculated. RESULTS: A total of 3260 clinical HER-2 tests by FISH were performed on 2963 serially referred breast cancer specimens. Of these, 2933 tests were successful and 2913 breast cancer specimens had both FISH and immunohistochemistry results available. With FISH as the standard testing method, the positive predictive value of positive immunohistochemistry score (3+) was 91.6%, and the negative predictive value of negative immunohistochemistry score (0 or 1+) was 97.2%. The sensitivity of immunohistochemistry tests, including tumor sections with scores of 2+ or 3+, was 92.6% and the specificity of immunohistochemistry tests with scores of 3+ was 98.8%. The FISH test had a significantly higher failure rate (5% vs 0.08%) and reagent cost (140 dollars vs 10 dollars), and longer testing (36 hours vs 4 hours) and interpretation times (7 minutes vs 45 seconds) vs immunohistochemistry tests. CONCLUSIONS: A testing algorithm for HER-2 determination is most efficient by using immunohistochemistry as the method of choice, with FISH performed for cancers with indeterminate results (2+ score). Successful quality control and quality assurance programs are a prerequisite for such approaches. PMID- 15113816 TI - Biomarkers of endothelial dysfunction and risk of type 2 diabetes mellitus. AB - CONTEXT: Endothelial dysfunction occurs in diagnosed type 2 diabetes mellitus but may also precede development of diabetes. OBJECTIVE: To determine whether elevated plasma levels of biomarkers reflecting endothelial dysfunction (E selectin; intercellular adhesion molecule 1 [ICAM-1]; and vascular cell adhesion molecule 1 [VCAM-1]) predict development of type 2 diabetes in initially nondiabetic women. DESIGN AND SETTING: Prospective, nested case-control study within the Nurses' Health Study, an ongoing US study initiated in 1976. PARTICIPANTS: Of 121 700 women initially enrolled, 32 826 provided blood samples in 1989-1990; of those free of diabetes, cardiovascular disease, or cancer at baseline, 737 developed incident diabetes by 2000. Controls (n = 785) were selected according to matched age, fasting status, and race. MAIN OUTCOME MEASURE: Risk of confirmed clinically diagnosed type 2 diabetes by baseline levels of E-selectin, ICAM-1, and VCAM-1. RESULTS: Baseline median levels of the biomarkers were significantly higher among cases than among controls (E-selectin, 61.2 vs 45.4 ng/mL; ICAM-1, 264.9 vs 247.0 ng/mL; VCAM-1, 545.4 vs 526.0 ng/mL [all P values < or =.004]). Elevated E-selectin and ICAM-1 levels predicted incident diabetes in logistic regression models conditioned on matching criteria and adjusted for body mass index (BMI), family history of diabetes, smoking, diet score, alcohol intake, activity index, and postmenopausal hormone use. The adjusted relative risks for incident diabetes in the top quintile vs the bottom quintiles were 5.43 for E-selectin (95% confidence interval [CI], 3.47-8.50), 3.56 for ICAM-1 (95% CI, 2.28-5.58), and 1.12 for VCAM-1 (95% CI, 0.76-1.66). Adjustment for waist circumference instead of BMI or further adjustment for baseline levels of C-reactive protein, fasting insulin, and hemoglobin A(1c) or exclusion of cases diagnosed during the first 4 years of follow-up did not alter these associations. CONCLUSION: Endothelial dysfunction predicts type 2 diabetes in women independent of other known risk factors, including obesity and subclinical inflammation. PMID- 15113817 TI - Antepartum dental radiography and infant low birth weight. AB - CONTEXT: Both high- and low-dose radiation exposures in women have been associated with low-birth-weight offspring. It is unclear if radiation affects the hypothalamus-pituitary-thyroid axis and thereby indirectly birth weight, or if the radiation directly affects the reproductive organs. OBJECTIVE: To investigate whether antepartum dental radiography is associated with low-birth weight offspring. DESIGN: A population-based case-control study. PARTICIPANTS AND SETTING: Enrollees of a dental insurance plan with live singleton births in Washington State between January 1993 and December 2000. Cases were 1117 women with low-birth-weight infants (<2500 g), of whom 336 were term low-birth-weight infants (1501-2499 g and gestation > or =37 weeks). Four control pregnancies resulting in normal-birth-weight infants (> or =2500 g) were randomly selected for each case (n = 4468). MAIN OUTCOME MEASURES: Odds of low birth weight and term low birth weight by dental radiographic dose during gestation. RESULTS: An exposure higher than 0.4 milligray (mGy) during gestation occurred in 21 (1.9%) mothers of low-birth-weight infants and, when compared with women who had no known dental radiography, was associated with an adjusted odds ratio (OR) for a low-birth-weight infant of 2.27 (95% confidence interval [CI], 1.11-4.66, P =.03). Exposure higher than 0.4 mGy occurred in 10 (3%) term low-birth-weight pregnancies and was associated with an adjusted OR for a term low-birth-weight infant of 3.61 (95% CI, 1.46-8.92, P =.005). CONCLUSION: Dental radiography during pregnancy is associated with low birth weight, specifically with term low birth weight. PMID- 15113818 TI - One-year health assessment of adult survivors of Bacillus anthracis infection. AB - CONTEXT: Little is known about potential long-term health effects of bioterrorism related Bacillus anthracis infection. OBJECTIVE: To describe the relationship between anthrax infection and persistent somatic symptoms among adults surviving bioterrorism-related anthrax disease approximately 1 year after illness onset in 2001. DESIGN, SETTING, AND PARTICIPANTS: Cross-sectional study of 15 of 16 adult survivors from September through December 2002 using a clinical interview, a medical review-of-system questionnaire, 2 standardized self-administered questionnaires, and a review of available medical records. MAIN OUTCOME MEASURES: Health complaints summarized by the body system affected and by symptom categories; psychological distress measured by the Revised 90-Item Symptom Checklist; and health-related quality-of-life indices by the Medical Outcomes Study 36-Item Short-Form Health Survey (version 2). RESULTS: The anthrax survivors reported symptoms affecting multiple body systems, significantly greater overall psychological distress (P<.001), and significantly reduced health related quality-of-life indices compared with US referent populations. Eight survivors (53%) had not returned to work since their infection. Comparing disease manifestations, inhalational survivors reported significantly lower overall physical health than cutaneous survivors (mean scores, 30 vs 41; P =.02). Available medical records could not explain the persisting health complaints. CONCLUSION: The anthrax survivors continued to report significant health problems and poor life adjustment 1 year after onset of bioterrorism-related anthrax disease. PMID- 15113819 TI - Effect of Vitamin D on falls: a meta-analysis. AB - CONTEXT: Falls among elderly individuals occur frequently, increase with age, and lead to substantial morbidity and mortality. The role of vitamin D in preventing falls among elderly people has not been well established. OBJECTIVE: To assess the effectiveness of vitamin D in preventing an older person from falling. DATA SOURCES: MEDLINE and the Cochrane Controlled Trials Register from January 1960 to February 2004, EMBASE from January 1991 to February 2004, clinical experts, bibliographies, and abstracts. Search terms included trial terms: randomized controlled trial or controlled-clinical trial or random-allocation or double blind method, or single-blind method or uncontrolled-trials with vitamin D terms: cholecalciferol or hydroxycholecalciferols or calcifediol or dihydroxycholecalciferols or calcitriol or vitamin D/aa[analogs & derivates] or ergocalciferol or vitamin D/bl[blood]; and with accidental falls or falls, and humans. STUDY SELECTION: We included only double-blind randomized, controlled trials (RCTs) of vitamin D in elderly populations (mean age, 60 years) that examined falls resulting from low trauma for which the method of fall ascertainment and definition of falls were defined explicitly. Studies including patients in unstable health states were excluded. Five of 38 identified studies were included in the primary analysis and 5 other studies were included in a sensitivity analysis. DATA EXTRACTION: Independent extraction by 3 authors using predefined data fields including study quality indicators. DATA SYNTHESIS: Based on 5 RCTs involving 1237 participants, vitamin D reduced the corrected odds ratio (OR) of falling by 22% (corrected OR, 0.78; 95% confidence interval [CI], 0.64 0.92) compared with patients receiving calcium or placebo. From the pooled risk difference, the number needed to treat (NNT) was 15 (95% CI, 8-53), or equivalently 15 patients would need to be treated with vitamin D to prevent 1 person from falling. The inclusion of 5 additional studies, involving 10 001 participants, in a sensitivity analysis resulted in a smaller but still significant effect size (corrected RR, 0.87; 95% CI, 0.80-0.96). Subgroup analyses suggested that the effect size was independent of calcium supplementation, type of vitamin D, duration of therapy, and sex, but reduced sample sizes made the results statistically nonsignificant for calcium supplementation, cholecalciferol, and among men. CONCLUSIONS: Vitamin D supplementation appears to reduce the risk of falls among ambulatory or institutionalized older individuals with stable health by more than 20%. Further studies examining the effect of alternative types of vitamin D and their doses, the role of calcium supplementation, and effects in men should be considered. PMID- 15113820 TI - Sex differences in developmental reading disability: new findings from 4 epidemiological studies. AB - CONTEXT: An influential article published in 1990 claimed that the increased rate of reading disability in boys was a consequence of referral bias. OBJECTIVES: To summarize the history of research on sex differences in reading disability and to provide new evidence from 4 independent epidemiological studies about the nature, extent, and significance of sex differences in reading disability. DESIGN, SETTING, AND PARTICIPANTS: The Dunedin Multidisciplinary Health and Development Study comprised 989 individuals (52.1% male) in a cohort born between April 1972 and March 1973 in Dunedin, New Zealand, and followed up from age 3 years; reading performance and IQ were assessed at ages 7, 9, and 11 years using the Burt Word Reading Test and the Wechsler Intelligence Scale for Children-Revised (WISC-R), respectively. The Christchurch Health and Development Study comprised 895 individuals (50% male) in a prospectively studied cohort born in the Christchurch, New Zealand, region during a 4-month period in 1977; reading performance and IQ were assessed at ages 8 to 10 years using the Burt Word Reading Test and the WISC-R. The Office for National Statistics (ONS) Study comprised a UK nationally representative sample of 5752 children (50.1% male) aged 9 to 15 years in 1999; reading was assessed on the British Ability Scales II and IQ on the British Picture Vocabulary Scales II. The Environmental Risk Longitudinal Twin Study (E-Risk) comprised 2163 twin children from England and Wales (49.1% male) identified at birth in 1994 and 1995 and included administration of the Test of Word Reading Efficiency at age 7 years and the Wechsler Preschool and Primary Scale of Intelligence-Revised as a test of IQ at age 5 years. MAIN OUTCOME MEASURE: Reading performance by sex in the lowest 15% of the distribution for all 4 studies, with and without taking IQ into account. RESULTS: In all 4 studies, the rates of reading disability were significantly higher in boys. For non-IQ-referenced reading disability: Dunedin study, 21.6% in boys vs 7.9% in girls (odds ratio [OR], 3.19; 95% confidence interval [CI], 2.15 4.17); Christchurch study, 20.6% in boys vs 9.8% in girls (OR, 2.38; 95% CI, 1.62 3.50); ONS study, 17.6% in boys vs 13.0% in girls (OR, 1.43; 95% CI, 1.23-1.65); and E-Risk, 18.0% in boys vs 13.0% in girls (OR, 1.39; 95% CI, 1.04-1.86). The rates for IQ-referenced reading disabilities were similar. CONCLUSION: Reading disabilities are clearly more frequent in boys than in girls. PMID- 15113821 TI - Risk factors for obstructive sleep apnea in adults. PMID- 15113822 TI - Vasopressin receptor antagonists: will the "vaptans" fulfill their promise? PMID- 15113823 TI - High-quality HER-2 testing: setting a standard for oncologic biomarker assessment. PMID- 15113824 TI - JAMA patient page. Reading disorder. PMID- 15113825 TI - First determination of the inhibitor complex structure of human hematopoietic prostaglandin D synthase. AB - Hematopoietic prostaglandin (PG) D synthase (H-PGDS) is responsible for the production of PGD(2) as an allergy or inflammation mediator in mast and Th2 cells. We determined the X-ray structure of human H-PGDS complexed with an inhibitor, 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazidyl) tetrazolium chloride (BSPT) at 1.9 A resolution in the presence of Mg(2+). The styryl group of the inhibitor penetrated to the bottom of the active site cleft, and the tetrazole ring was stabilized by the stacking interaction with Trp104, inducing large movement around the alpha5-helix, which caused the space group of the complex crystal to change from P2(1) to P1 upon binding of BSPT. The phthalhydrazidyl group of BSPT exhibited steric hindrance due to the cofactor, glutathione (GSH), increasing the IC(50) value of BSPT for human H-PGDS from 36.2 micro M to 98.1 micro M upon binding of Mg(2+), because the K(m) value of GSH for human H-PGDS was decreased from 0.60 micro M in the presence of EDTA to 0.14 micro M in the presence of Mg(2+). We have to avoid steric hindrance of the GSH molecule that was stabilized by intracellular Mg(2+) in the mM range in the cytosol for further development of structure-based anti-allergic drugs. PMID- 15113826 TI - DNA display of biologically active proteins for in vitro protein selection. AB - In vitro display technologies are powerful tools for screening peptides with desired functions. We previously proposed a DNA display system in which streptavidin-fused peptides are linked with their encoding DNAs via biotin labels in emulsion compartments and successfully applied it to the screening of random peptide libraries. Here we describe its application to functional and folded proteins. By introducing peptide linkers between streptavidin and fused proteins, we achieved highly efficient (>95%) formation of DNA-protein conjugates. Furthermore, we successfully enriched a glutathione-S-transferase gene by a factor of 20-30-fold per round on glutathione-coupled beads. Thus, DNA display should be useful for rapidly screening or evolving proteins based on affinity selection. PMID- 15113827 TI - A novel mechanism of intragenic complementation between Phe to Ala calmodulin mutations. AB - Calmodulin (CaM) performs essential functions in cell proliferation in Saccharomyces cerevisiae. Previously, we isolated fourteen temperature-sensitive Phe-to-Ala mutations of the CaM-encoding gene CMD1. These mutations were classified into four intragenic complementation groups, suggesting that each group represents a loss of CaM interaction with its specific essential target protein. Nuf1p/Spc110p, one of the essential targets, is a spindle pole body component that is required for proper mitosis. We investigated which intragenic complementation group of CaM represents the malfunction of Nuf1p. Immunoprecipitation analysis showed that two cmd1 mutations belonging to two distinct intragenic complementation groups had the most severely impaired complex formation with Nuf1p at the restrictive temperature. The temperature-sensitive growth of these cmd1 mutants was suppressed by a CaM-independent dominant allele of NUF1. Additionally, these mutants displayed characteristic mitotic defects: an increased ratio of artificial chromosome loss, which could be suppressed by the CaM-independent dominant allele of NUF1, and aberrant microtubule structures. These results indicate that these cmd1 mutants display the temperature-sensitive growth due to the compromised interaction with Nuf1p. However, the interaction was restored in a heterozygous diploid of the two cmd1 alleles, suggesting that intragenic complementation between these cmd1 alleles occurs by a novel mechanism, whereby co-presence of both mutant proteins rescues the interaction with Nuf1p. PMID- 15113828 TI - Perinerin, a novel antimicrobial peptide purified from the clamworm Perinereis aibuhitensis grube and its partial characterization. AB - A novel antimicrobial peptide was isolated and partially characterized from the homogenate of an Asian marine clamworm, Perinereis aibuhitensis Grube. This novel peptide, named Perinerin, was purified to homogeneity by heparin-affinity column and reverse-phase HPLC, and biologically tested with a MTS-PMS colorimetric assay. Perinerin consists of 51 amino acid residues and structurally appears to be highly basic and hydrophobic. It shows marked activity in vitro against both Gram-negative and Gram-positive bacteria and fungi, which indicates a bactericidal effect as well. Perinerin appears to be constitutively present and its sequence is novel among all other known antimicrobial peptides. These results suggest that Perinerin has the potential to serve as a convenient "evaluation marker" for studying alterations in the biochemistry of the host, particularly with respect to environmental changes. In addition, the MTS-PMS colorimetric assay examination of antimicrobial activity appears to be superior to existing methods and may offer more general application in the search for new antibiotic molecules. PMID- 15113829 TI - Molecular breeding of 2,3-dihydroxybiphenyl 1,2-dioxygenase for enhanced resistance to 3-chlorocatechol. AB - 3-Chlorobiphenyl is known to be mineralized by biphenyl-utilizing bacteria to 3 chlorobenzoate, which is further metabolized to 3-chlorocatechol. An extradiol dioxygenase, 2,3-dihydroxybiphenyl 1,2-dioxygenase (DHB12O; EC 1.13.11.39), which is encoded by the bphC gene, catalyzes the third step of the upper pathway of 3 chlorobiphenyl degradation. In this study, two full-length bphCs and nine partial fragments of bphCs fused to the 3' end of bphC in Pseudomonas pseudoalcaligenes KF707 were cloned from different biphenyl-utilizing soil bacteria and expressed in Escherichia coli. The enzyme activities of the expressed DHB12Os were inhibited to varying degrees by 3-chlorocatechol, and the E. coli cells overexpressing DHB12O could not grow or grew very slowly in the presence of 3 chlorocatechol. These sensitivities of enzyme activity and cell growth to 3 chlorocatechol were well correlated, and this phenomenon was employed in screening chimeric BphCs formed by family shuffling of bphC genes isolated from Comamonas testosteroni KF704 and C. testosteroni KF712. The resultant DHB12Os were more resistant by a factor of two to 3-chlorocatechol than one of the best parents, KF707 DHB12O. PMID- 15113830 TI - Production of N-lauroylated G protein alpha-subunit in Sf9 insect cells: the type of N-acyl group of Galpha influences G protein-mediated signal transduction. AB - The alpha-subunit of rod photoreceptor G protein transducin (G(t1)alpha) is heterogeneously modified at the N-terminus by a mixture of acyl groups, laurate (C12:0), myristate (C14:0), and two unsaturated fatty acids (C14:1 and C14:2). Although the N-fatty acylation of G(t1)alpha plays important roles in protein protein and protein-membrane interactions in light signaling, the biological significance of the heterogeneous acylation remains unclear due to the difficulty in isolating each G(t1)alpha isoform from the retinal rod cells. Here we found that G(t1)alpha/G(i1)alpha chimera (G(t/i)alpha) expressed in Sf9 cells is also heterogeneously modified by myristate (approximately 90%) and laurate (approximately 10%), raising the possibility that the N-acyl group of recombinant G(t/i)alpha may be manipulated by modifying culture media. In fact, addition of myristic acid to the medium decreased the relative content of lauroylated G(t/i)alpha to an undetectable level, whereas exogenously added lauric acid significantly increased the relative content of lauroylated G(t/i)alpha in a concentration-dependent manner. By culturing the G(t/i)alpha-virus infected Sf9 cells with fatty acids, we obtained four different preparations of N-acylated G(t/i)alpha, in which the relative abundance of lauroylated isoform was 0%, 20%, 33% and approximately 70%, respectively. Functional analysis of these proteins showed that an increase in the relative content of the lauroylated isoform remarkably slowed down the steady-state GTP hydrolysis rate of G(t/i)alpha; the steady-state GTPase activity of the lauroylated isoform was estimated to be one order of magnitude lower than that of the myristoylated isoform. These results suggest that the retinal G(t1)alpha is composed of isoforms having functionally heterogeneous signaling properties. PMID- 15113831 TI - Overexpression of DRG2 increases G2/M phase cells and decreases sensitivity to nocodazole-induced apoptosis. AB - DRG2, a member of the DRG subfamily in the GTP-binding protein superfamily, was identified as a repressed gene product in fibroblasts transformed by SV40. The significance of this down-regulation and the cellular role of DRG2 has not been understood in the past. To investigate the function of DRG2 we made a Jurkat cell line, Jurkat-LNCX2-DRG2, stably transfected with pLNCX2-DRG2 to overexpress human DRG2. Cell cycle distribution analysis revealed an increased accumulation of G(2)/M phase cells in Jurkat-LNCX2-DRG2 cells, indicating a retardation of cell cycle progression. In addition, an overexpression of DRG2 reduced the sensitivity of Jurkat cells to the mitotic poison nocodazole. Our data suggest that overexpression of DRG2 in Jurkat cells affects genes regulating cell-cycle arrest and apoptosis, and that these molecular changes may be important in the growth or differentiation of cells. PMID- 15113832 TI - pH-induced conformational transition of H. pylori acyl carrier protein: insight into the unfolding of local structure. AB - Acyl carrier protein (ACP) is a small acidic protein and its primary structure is highly conserved in various bacterial sources. Despite its small size, it interacts with diverse proteins associated with many biosynthetic pathways. The three-dimensional structure of H. pylori ACP and its structural characteristics were clarified using NMR and CD spectroscopy. H. pylori ACP consists of four helices connected by different sized loops. The helices correspond to residues L3 Q14 (alphaI), S36-G50 (alphaII), D56-E60 (alphaIII), and V65-K76 (alphaVI). The size of each helix differs slightly from that of homologous ACPs. However, H. pylori ACP showed a distinct pH-dependent conformational characteristic: at neutral pH, it adopts a partially unfolded structure, while it has a tight fold at pH 6. The chemical shift perturbation and (1)H-(15)N steady state NOE analysis at both pH 6 and 7 showed that the local change of structural components occurred mainly around loop II, and this change was reflected by the changes of the residues Ile 54 and Asp 56. Examination of the structure showed that the network of Glu 47, Ile 54, Asn 75, and Lys 76 is very important for the structural stability. The pH-dependent folding process shows a kind of cooperativity, since all the residues involved in the conformational transitions are contiguous and in spatial proximity. PMID- 15113833 TI - Module-specific antibodies against human connective tissue growth factor: utility for structural and functional analysis of the factor as related to chondrocytes. AB - Connective tissue growth factor/hypertrophic chondrocyte specific gene product 24 (CTGF/Hcs24/CCN2) shows diverse functions in the process of endochondral ossification. It promotes not only the proliferation and differentiation of chondrocytes and osteoblasts in vitro, but also angiogenesis in vivo. The ctgf gene is a member of the gene family called CCN, and it encodes the characteristic 4-module structure of this family, with the protein containing IGFBP, VWC, TSP and CT modules. We raised several monoclonal antibodies and polyclonal antisera against CTGF, and located the epitopes in the modules by Western blotting. For mapping the epitopes, Brevibacillus-produced independent modules were utilized. As a result, at least 1 antibody or antiserum was prepared for the detection of each module in CTGF. Western blotting with these antibodies is expected to be useful for the analysis of CTGF fragmentation. Moreover, we examined the effects of these monoclonal antibodies on the biological functions of CTGF. One out of 3 humanized monoclonal antibodies was found to neutralize efficiently the stimulatory effect of CTGF on chondrocytic cell proliferation. This particular antibody bound to the CT module. In contrast, surprisingly, all of the 3 antibodies recognizing IGFBP, VWC and CT modules stimulated proteoglycan synthesis in chondrocytic cells. Together with previous findings, these results provide insight into the structural-functional relationships of CTGF in executing multiple functions. PMID- 15113834 TI - Two additional carbohydrate-binding sites of beta-amylase from Bacillus cereus var. mycoides are involved in hydrolysis and raw starch-binding. AB - In the previous X-ray crystallographic study, it was found that beta-amylase from Bacillus cereus var. mycoides has three carbohydrate-binding sites aside from the active site: two (Site2 and Site3) in domain B and one (Site1) in domain C. To investigate the roles of these sites in the catalytic reaction and raw starch binding, Site1 and Site2 were mutated. From analyses of the raw starch-binding of wild-type and mutant enzymes, it was found that Site1 contributes to the binding affinity to raw-starch more than Site2, and that the binding capacity is maintained when either Site1 or Site2 exists. The raw starch-digesting ability of this enzyme was poor. From inhibition studies by maltitol, GGX and alpha-CD for hydrolyses of maltopentaose (G5) and amylose ( (n) = 16) catalyzed by wild-type and mutant enzymes, it was found that alpha-CD is a competitive inhibitor, while, maltitol behaves as a mixed-type or competitive inhibitor depending on the chain length of the substrate and the mutant enzyme. From the analysis of the inhibition mechanism, we conclude that the bindings of maltitol and GGX to Site2 in domain B form an abortive ESI complex when amylose ( (n) = 16) is used as a substrate. PMID- 15113835 TI - Molecular identification of K-CL cotransporter in dog erythroid progenitor cells. AB - KCC1 cDNA was cloned in dog erythroblasts that had differentiated from peripheral mononuclear cells. The size of the cDNA was 3,258 bp, the same as in pigs, but 3 bp longer than in humans and rodents. The dog KCC1 cDNA encodes for 1,086 amino acid residues with a calculated molecular mass of 120 kDa. The 560 bp cDNA fragment from position 679 to 1,238 in the full length cDNA from the dog erythroblasts was 100% identical to that in the kidney. Hydropathy analysis showed that the structure of dog KCC1 was similar to in other species; 12 trans membrane domains, four glycosylation sites in loop 5, and 17 consensus phosphorylation sites in the cytosol. However, there were variations in dog KCC1 compared to in other species; there was one CK2 phosphorylation site that was found only in dog KCC1. There were also substitutions of amino acids that affect pH sensitivity (His) and change acidic/basic residues or charged residues. In HEK 293 cells transfected with dog KCC1 cDNA (HEK-dKCC1), the Rb influx, which was ouabain-resistant, Cl-dependent, N-ethyl maleimide (NEM)- stimulative and Na independent, was measured as for K-Cl cotransport, and the influx was found to be increased approximately 3 fold in HEK-dKCC1 compared to in the control. This ouabain-resistant Cl-dependent Rb influx was also volume-sensitive in hyposmotic medium, and the volume-sensitive component was inhibited by furosemide. Thus, the KCC1 cDNA cloned in dog erythroblasts encodes a volume-sensitive K-Cl cotransporter. PMID- 15113836 TI - Thermus thermophilus MutS2, a MutS paralogue, possesses an endonuclease activity promoted by MutL. AB - The mismatch repair system (MMR) recognizes and corrects mismatched or unpaired bases caused mainly by DNA polymerase, and contributes to the fidelity of DNA replication in living cells. In Escherichia coli, the MutHLS system is known to function in MMR, and homologues of MutS and MutL are widely conserved in almost all organisms. However, the MutH endonuclease has not been found in the majority of organisms. Such organisms, including Thermus thermophilus HB8, often possess the so-called MutS2 protein, which is highly homologous to MutS but contains an extra C-terminal stretch. To elucidate the function of MutS2, we overexpressed and purified T. thermophilus MutS2 (ttMutS2). ttMutS2 demonstrated the ability to bind double-stranded (ds) DNA, but, unlike ttMutS, ttMutS2 showed no specificity for mismatched duplexes. ttMutS2 ATPase activity was also detected and was stimulated by dsDNA. Our results also showed that ttMutS2 incises dsDNA. ttMutS2 incises not only oligo dsDNA but also plasmid DNA, suggesting that ttMutS2 possesses an endonuclease activity. At low concentrations, the incision activity was not retained, but was promoted by T. thermophilus MutL. PMID- 15113837 TI - Association with Hrs is required for the early endosomal localization, stability, and function of STAM. AB - Members of the STAM family of proteins, STAM1 and STAM2, are associated with Hrs through their coiled-coil regions. Both Hrs and STAM bind ubiquitin and are involved in endosomal sorting of ubiquitinated cargo proteins for trafficking to the lysosome. Here we examined the biological significance of STAM binding to Hrs. Endogenous STAM1 and STAM2 were mostly localized on the early endosome, suggesting that they are resident endosomal proteins. A STAM2 mutant that lacks the coiled-coil region and does not bind Hrs, in contrast, mislocalized to the cytoplasm. Deletion of a region located N-terminal to the coiled-coil region and conserved among STAM proteins also severely affected Hrs binding and the endosomal localization of STAM2, suggesting that this region is also involved in these activities. Depletion of endogenous Hrs by RNA interference similarly caused the mislocalization of exogenously expressed STAM2 to the cytoplasm. These results indicate that STAM is localized to the early endosome by binding to Hrs on the target membrane. In addition, the expression level of endogenous STAM proteins was drastically reduced in Hrs-depleted cells, suggesting that STAM is stabilized by binding to Hrs. Finally, STAM2 mutants lacking the Hrs-binding activity were defective in causing the enlargement of early endosomes, accumulating ubiquitinated proteins on this aberrant organelle, and inhibiting the degradation of ligand-activated epidermal growth factor receptors, suggesting that the association with Hrs is a prerequisite for STAM function. PMID- 15113838 TI - Characterization of bacteriophage T3 DNA ligase. AB - DNA ligases of bacteriophage T4 and T7 have been widely used in molecular biology for decades, but little is known about bacteriophage T3 DNA ligase. Here is the first report on the cloning, expression and biochemical characterization of bacteriophage T3 DNA ligase. The polyhistidine-tagged recombinant T3 DNA ligase was shown to be an ATP-dependent enzyme. The enzymatic activity was not affected by high concentration of monovalent cations up to 1 M, whereas 2 mM ATP could inhibit its activity by 50%. Under optimal conditions (pH 8.0, 0.5 mM ATP, 5 mM DTT, 1 mM Mg(2+) and 300 mM Na(+)), 1 fmol of T3 DNA ligase could achieve 90% ligation of 450 fmol of cohesive dsDNA fragments in 30 min. T3 DNA ligase was shown to be over 5-fold more efficient than T4 DNA ligase for ligation of cohesive DNA fragments, but less active for blunt-ended DNA fragments. Phylogenetic analysis showed that T3 DNA ligase is more closely related to T7 DNA ligase than to T4 DNA ligase. PMID- 15113839 TI - Secondary-structure analysis of proteins by vacuum-ultraviolet circular dichroism spectroscopy. AB - The vacuum ultraviolet circular dichroism (VUVCD) spectra of 15 globular proteins (myoglobin, hemoglobin, human serum albumin, cytochrome c, peroxidase, alpha lactalbumin, lysozyme, ovalbumin, ribonuclease A, beta-lactoglobulin, pepsin, trypsinogen, alpha-chymotrypsinogen, soybean trypsin inhibitor, and concanavalin A) were measured in aqueous solutions at 25 degrees C in the wavelength region from 260 to 160 nm under a high vacuum, using a synchrotron-radiation VUVCD spectrophotometer. The VUVCD spectra below 190 nm revealed some characteristic bands corresponding to different secondary structures. The contents of alpha helices, beta-strands, turns, and unordered structures were estimated using the SELCON3 program with VUVCD spectra data on the 15 proteins. Prediction of the secondary-structure contents was greatly improved by extending the circular dichroism spectra to 165 nm. The numbers of alpha-helix and beta-strand segments calculated from the distorted alpha-helix and beta-strand contents did not differ greatly from those obtained from X-ray crystal structures. These results demonstrate that synchrotron-radiation VUVCD spectroscopy is a powerful tool for analyzing the secondary structures of proteins. PMID- 15113840 TI - Distal Sox binding elements of the alphaB-crystallin gene show lens enhancer activity in transgenic mouse embryos. AB - alphaB-Crystallin, a member of the small heat shock protein (sHSP) family, is expressed in various tissues including lens, heart, and skeletal muscle. Previously we identified the gene of HSPB2, another member of the sHSP family, located 1-kb upstream of the alphaB-crystallin gene in a head-to-head manner. In the present study, we found a highly conserved region of 220 bp approximately 2.4 kb upstream of the alphaB-crystallin gene and examined its role in expression of the alphaB-crystallin gene. Transgenic mice containing 3 kb of the upstream sequence of the alphaB-crystallin gene showed lacZ reporter gene expression in the lens as well as the myotome and heart on embryonic day 12.5. Deletion analysis revealed that the -2656/-2267 region including the conserved region with four putative Sox binding elements (E1-E4) exhibits lens enhancer activity toward the alphaB-crystallin promoter. Gel shift assays showed that the Sox1 and Sox2 proteins preferentially bound to E2 and E4. Moreover, disruption of E2 and E4 abolished the reporter gene expression in the lens. These results indicate that the newly identified enhancer with Sox elements activates the alphaB-crystallin promoter in the lens, although they are separated by the entire HSPB2 gene. PMID- 15113841 TI - Inhibitory effect of 0.19 alpha-amylase inhibitor from wheat kernel on the activity of porcine pancreas alpha-amylase and its thermal stability. AB - The inhibitory effect of 0.19 alpha-amylase inhibitor (0.19 AI) from wheat kernel on the porcine pancreas alpha-amylase (PPA)-catalyzed hydrolysis of p-nitrophenyl alpha-D-maltoside (pNP-G2) was examined. 0.19 AI is a homodimer of 26.6 kDa with 13.3-kDa subunits under the conditions used. The elution behaviors in gel filtration HPLC of PPA and 0.19 AI indicated that a PPA molecule bound with a 0.19 AI molecule (homodimer) at a molar ratio of 1:1. 0.19 AI inhibited PPA activity in a competitive manner with an inhibitor constant, K(i), of 57.3 nM at pH 6.9, 30 degrees C, and the binding between them was found to be endothermic and entropy-driven. The activation energy for the thermal inactivation of 0.19 AI was determined to be 87.0 kJ/mol, and the temperature, T(50), giving 50% inactivation in a 30-min incubation at pH 6.9 was 88.1 degrees C. The high inhibitory activity of 0.19 AI against PPA and its high thermal stability suggest its potential for use in the prevention and therapy of obesity and diabetes. PMID- 15113842 TI - The carboxyl-terminus directs TAF(I)48 to the nucleus and nucleolus and associates with multiple nuclear import receptors. AB - The protein complex Selectivity Factor 1, composed of TBP, TAF(I)48, TAF(I)63 and TAF(I)110, is required for rRNA transcription by RNA polymerase I in the nucleolus. The steps involved in targeting Selectivity Factor 1 will be dependent on the transport pathways that are used and the localization signals that direct this trafficking. In order to investigate these issues, we characterized human TAF(I)48, a subunit of Selectivity Factor 1. By domain analysis of TAF(I)48, the carboxyl-terminal 51 residues were found to be required for the localization of TAF(I)48, as well as sufficient to direct Green Fluorescent Protein to the nucleus and nucleolus. The carboxyl-terminus of TAF(I)48 also has the ability to associate with multiple members of the beta-karyopherin family of nuclear import receptors, including importin beta (karyopherin beta1), transportin (karyopherin beta2) and RanBP5 (karyopherin beta3), in a Ran-dependent manner. This property of interacting with multiple beta-karyopherins has been previously reported for the nuclear localization signals of some ribosomal proteins that are likewise directed to the nucleolus. This study identifies the first nuclear import sequence identified within the TBP-Associated Factor subunits of Selectivity Factor 1. PMID- 15113843 TI - Nerve growth factor attenuates endoplasmic reticulum stress-mediated apoptosis via suppression of caspase-12 activity. AB - Following endoplasmic reticulum (ER) stress, which occurs via inhibition of the glycosylation of newly synthesized proteins, caspase family proteins are activated to promote ER stress-mediated apoptosis. Here we report that nerve growth factor (NGF) suppressed the ER stress-mediated apoptosis in tunicamycin treated PC12 cells through an extensive decrease of the caspase-3/-9/-12 activity. Detailed analysis of the mechanism underlying the NGF-mediated cell survival revealed that the activities of all seriate caspases were reduced through the phosphatidylinositol 3-kinase (PI3-K) signaling pathway induced by NGF. Moreover, we found that the activity of c-Jun N-terminal kinase (JNK) was not essential for the tunicamycin-induced apoptosis of PC12 cells. These results demonstrate that the inactivation of caspase-12 via the NGF-mediated PI3-K signaling pathway leads to inactivation of the caspase cascade including caspase 3 and -9. PMID- 15113844 TI - Blocking of human immunodeficiency virus type-1 virion autolysis by autologous p2(gag) peptide. AB - Our previous study suggested that the p2(gag) peptide, AEAMSQVTNTATIM, inhibits human immunodeficiency virus type 1 (HIV-1) protease (PR) activity in vitro. In this study, Ala substitutions (Met4Ala and Thr8Ala) and deletion of amino acid Asn9 within the nona p2(gag) peptide (AEAMSQVTN) were found to decrease the inhibitory effect on HIV-1 PR activity. Furthermore, treatment of PMA-activated latently infected T lymphocytes, ACH-2 cells, with the p2(gag) peptide (100 and 250 micro M) resulted in a decrease in the amount of p24(gag )in the resultant viral lysates derived from the cell-free supernatant. In addition, the HIV-1-Tat p2(gag) fusion peptide was synthesized to effectively deliver the p2(gag) peptide into the cells. The fusion peptide was incorporated into chronically infected T lymphocytes, CEM/LAV-1 cells, as detected on indirect immunofluorescence analysis using anti-p2(gag) peptide monoclonal antibodies, which recognize the nona peptide (AEAMSQVTN) derived from the N-terminus of the p2(gag) peptide, and cleaved by HIV-1 PR in vitro. Treatment of CEM/LAV-1 cells with the fusion peptide also resulted in a decrease in the amount of p24(gag )in the resultant viral lysate derived from the cell-free supernatant. Taken together, these data suggest that the p2(gag) peptide consequently blocks the autolysis of HIV-1 virions for the conservation of viral species. PMID- 15113845 TI - Ligand dependency of 5-hydroxytryptamine 2C receptor internalization. AB - Agonist-induced internalization of G protein-coupled receptors (GPCRs) is a well characterized phenomenon believed to contribute to receptor desensitization. The 5-hydroxytryptamine (5-HT)2C subtype of serotonin receptor is a GPCR that we have shown to internalize upon agonist incubation. In this study, we have examined the effects of 5-HT2C receptor agonists serotonin, Ro 60-0175 [(S)-2-(6-chloro-5 fluoroindol-1-yl)-1-methylethylamine], and WAY-161503 [(4aR)-8,9-dichloro 2,3,4,4a-tetrahydro-1H-pyrazino[1,2-a]quinoxalin-5(6H)-one]; partial agonists mCPP [1-(m-chlorophenyl)piperazine] and DOI [(+)-1-(2,5-dimethoxy-4-iodophenyl)-2 amino-propane]; inverse agonists SB-206553 [N-3-pyridinyl-3,5-dihydro-5 methylbenzo(1,2-b:4,5-b')dipyrrole-1(2H)carboxamide] and mianserin; and neutral antagonists SB-242084 [6-chloro-5-methyl-1-[[2-[(2-methyl-3-pyridyl)oxy]-5 pyridyl]carbamoyl]-indoline] and 5-methoxygramine on the internalization of a C terminal green fluorescent protein (GFP)-tagged 5-HT2C receptor (VSV isoform) expressed in transiently transfected human embryonic kidney cells. We detected internalization with an automated, cell-based fluorescence-imaging system (Arrayscan) and monitored function with intracellular Ca2+ measurements (flourometric imaging plate reader). The 5-HT2C-GFP construct exhibited appropriate pharmacology, and we observed that although all three agonists resulted in similar magnitudes of dose-dependent internalization, the partial agonists resulted in approximately 50% less internalization, and the inverse agonists and neutral antagonists failed to induce internalization. These results were confirmed by confocal microscopy. They demonstrate that the 5-HT2C receptor is internalized by incubation with agonists and partial agonists but not with inverse agonists or neutral antagonists. PMID- 15113846 TI - The distinct alterations produced in cardiovascular functions by prednisolone and nitro-prednisolone (NCX-1015) in the rat highlight a causal role for endothelin 1. AB - Daily administration of prednisolone, but not the derivative NCX-1015 (or prednisolone 21-[4'-nitrooxymethyl]benzoate), to rats resulted in a time- and dose-dependent increase in mean arterial blood pressure (MABP), significant after 1 week for the dose of 6.9 micromol/kg i.p. (n = 10; P < 0.05), and 3 weeks for the lower dose of 1.38 micromol/kg. A similar dichotomy of behavior was observed with respect to myocardial contractility and renal vascular resistance, in either case augmented by 3-week treatment with prednisolone but not NCX-1015. In contrast, both NCX-1015 and prednisolone reduced plasma levels of corticosterone in a dose- (dose range of 0.69-6.9 micromol/kg i.p.) and time-dependent (1-3 weeks) manner. Similar profiles were obtained for plasma nitrate values, although they were increased selectively after NCX-1015 administration. In contrast, prednisolone, but not NCX-1015, augmented plasma endothelin 1 (ET-1) with a profile that mirrored the changes observed in MABP and renal blood flow. Supply in the drinking water of the ET-1 receptor type A (ETA) antagonist FR139317 [(R-2 [(R)-2-[(S)-2-[[1-(hexahydro-1H-azepinyl)]-carbonyl]amino-4-methylpentanoyl] amino-3-(2-pyridil)propionic] or mixed ETA/B, but not of selective ETB, antagonists prevented the changes produced by a 21-day treatment with prednisolone. In conclusion, this study indicates 1) a lack of occurrence of cardiovascular alterations by nitro-releasing derivative of prednisolone (NCX 1015), and 2) a functional link between prednisolone effects and the endogenous endothelin-1 system. PMID- 15113847 TI - A new poly(ADP-ribose) polymerase inhibitor, FR261529 [2-(4-chlorophenyl)-5 quinoxalinecarboxamide], ameliorates methamphetamine-induced dopaminergic neurotoxicity in mice. AB - Methamphetamine (METH) administration in mice, results in a chronic dopamine (DA) depletion associated with nerve terminal damage, with DA oxidation and generation of reactive oxygen species (ROS) primarily mediating this neurotoxicity. The oxidative stress induced by METH putatively activates nuclear enzyme poly(ADP ribose) polymerase (PARP), with excessive PARP activation eventually leading to cell death. In this study, we show that prevention of PARP activation by treatment with FR261529 [2-(4-chlorophenyl)-5-quinoxalinecarboxamide], the compound that was recently identified as a novel PARP inhibitor (IC50 for PARP-1 = 33 nM, IC50 for PARP-2 = 7 nM), protects against both ROS-induced cells injury in vitro and METH-induced dopaminergic neuronal damage in an in vivo Parkinson's disease (PD) model. In PC12 cells, exposure of hydrogen peroxide or METH markedly induced PARP activation, and treatment with FR261529 (1 microM) significantly reduced PARP activation and attenuated cell death. In the mouse METH model, METH (15 mg/kg x 2 i.p., 2 h apart) intoxication accelerated DA metabolism and oxidation in the striatum, with subsequent cell damage in nigrostriatal dopaminergic neurons after 4 days. Oral administration of FR261529 (10 or 32 mg/kg) attenuated the damage of dopaminergic neurons via marked reduction of PARP activity and not via changes in dopamine metabolism or body temperature. These findings indicate that the neuroprotective effects of a novel PARP inhibitor, FR261529, were accompanied by inhibition of METH-induced PARP activation, suggesting that METH induces nigrostriatal dopaminergic neurodegeneration involving PARP activation and also orally active and brain-penetrable PARP inhibitor FR261529 could be a novel attractive therapeutic candidate for neurodegenerative disorders such as PD. PMID- 15113848 TI - Behavioral characterization of the novel GABAB receptor-positive modulator GS39783 (N,N'-dicyclopentyl-2-methylsulfanyl-5-nitro-pyrimidine-4,6-diamine): anxiolytic-like activity without side effects associated with baclofen or benzodiazepines. AB - The role of GABAB receptors in various behavioral processes has been largely defined using the prototypical GABAB receptor agonist baclofen. However, baclofen induces sedation, hypothermia and muscle relaxation, which may interfere with its use in behavioral paradigms. Although there is much evidence for a role of the inhibitory neurotransmitter GABA in the pathophysiology of anxiety, the role of GABAB receptors in these disorders is largely unclear. We recently identified GS39783 (N,N'-dicyclopentyl-2-methylsulfanyl-5-nitro-pyrimidine-4,6-diamine) as a selective allosteric positive modulator at GABAB receptors. The aim of the present study was to broadly characterize the effects of GS39783 in well validated rodent models for motor activity, cognition, and anxiety. The following tests were included: locomotor activity in rats and mice, rotarod and traction tests (including determinations of core temperature) in mice, passive avoidance in mice and rats, elevated plus maze in rats, elevated zero maze in mice and rats, stress-induced hyperthermia in mice, and pentobarbital- and ethanol-induced sleep in mice. Unlike baclofen and/or the benzodiazepine chlordiazepoxide, GS39783 had no effect in any of the tests for locomotion, cognition, temperature, or narcosis. Most interestingly, GS39783 had anxiolytic-like effects in all the tests used. Overall, the data obtained here suggest that positive modulation of GABAB receptors may serve as a novel therapeutic strategy for the development of anxiolytics, with a superior side effect profile to both baclofen and benzodiazepines. PMID- 15113849 TI - HPV infections and tonsillar carcinoma. AB - Since human papillomavirus (HPV) was first linked to laryngeal/oral carcinomas in 1983, several studies have confirmed its causal role in a subgroup of upper aerodigestive tract tumours. Of the non-genital cancers, tonsillar carcinomas (TCs) have the strongest association with HPV. By the end of 2002, 432 TCs had been analysed for HPV DNA. Overall detection rate was 51%, with HPV-16 being the most prevalent (84%). The original proposal that HPV-33 would be the most frequent HPV in TCs has not been confirmed, being present in only 4.6% of cases. HPV copy numbers are similar to those found in genital carcinomas (10-300 copies/cell), although HPV is mainly episomal in TC. The importance of this observation is unclear, although a role for subepithelial proliferative lymphatic tissue has been speculated. Patients with HPV-16 positive tumours have better overall and disease specific survival than HPV negative patients. They are also younger and the association with conventional risk factors-smoking and drinking is less significant than in HPV negative patients. Thus, recent data suggest a distinct pattern for HPV-16 positive TCs. PMID- 15113850 TI - MUC1 and MUC2 in pancreatic neoplasia. AB - MUCs are glycoproteins with various roles in homeostasis and carcinogenesis. Among other actions, MUC1 may inhibit cell-cell and cell-stroma interactions and function as a signal transducer, participating in cancer progression. In contrast, MUC2 is normally found only in goblet cells, where it contributes to the protective barrier function of these cells. Recently, a tumour suppressor role has been demonstrated for MUC2, and both MUC1 and MUC2 appear to have important roles in pancreatic neoplasia. MUC1 appears to be a marker of aggressive phenotype and may facilitate the vascular spread of carcinoma cells. In contrast, MUC2 is rarely detectable in aggressive pancreatic tumours, but is commonly expressed in intraductal papillary mucinous neoplasms (IPMNs), which are rare, indolent tumours, in intestinal IPMNs, and in indolent colloid carcinomas. MUC2 appears to be not only a marker of this indolent pathway, but also partly responsible for its less aggressive nature. Thus, in pancreatic neoplasia, MUC1 and MUC2 have potential diagnostic and prognostic value as markers of aggressive and indolent phenotypes, respectively, and have potential as therapeutic targets. PMID- 15113851 TI - KIT expression in fetal, normal adult, and neoplastic renal tissues. AB - BACKGROUND: KIT is a transmembrane tyrosine kinase receptor, expressed in high amounts in various normal cells. In addition, c-kit mutation or activation is a major pathogenetic event in certain tumours (such as gastrointestinal stromal tumours). There are only limited data in the literature on the expression of KIT in normal and neoplastic renal tissues. AIMS: To investigate KIT expression in normal and neoplastic renal tissues. METHODS: KIT expression was evaluated by means of immunohistochemistry in paraffin wax embedded sections from 67 tissue samples. RESULTS: Eight of eight fetal kidneys, and 10 of 10 normal adult kidneys revealed cytoplasmic staining of renal tubules. The three cases of renal dysplasia studied expressed KIT in their normal and aberrant tubules. Two of 13 conventional renal cell carcinomas (RCCs), two of seven papillary type RCCs, four of seven chromophobe type RCCs, none of six nephroblastomas, seven of seven oncocytomas, two of two mesoblastic nephromas, and two of four angiomyolipomas were positive. CONCLUSION: KIT is expressed in normal fetal and adult renal tubules, and in a subset of renal tumours. The expression of KIT in these renal tumours may prove to have diagnostic relevance and/or therapeutic implications. PMID- 15113852 TI - A model for determining the optimum histology of sentinel lymph nodes in breast cancer. AB - AIMS: To create and use a geometrical model for sentinel lymph node (SLN) histopathology in breast cancer. METHODS: The model involves a spherical metastasis randomly situated in an SLN. Two extreme situations are taken as the starting points. In one of these, the metastasis is seen in its largest dimension, whereas in the other it is only just visible, approximating 0 mm in size. Intermediate positions are analysed, with different metastasis sizes and different distances between the levels assessed by histology. RESULTS: The findings suggest that sections taken 1 mm apart afford a reasonable means of identifying almost all metastases measuring > 2 mm (referred to as macrometastases here). For nearly all micrometastases to be identified correctly according to the current TNM definitions (that is, metastases > 0.2 mm), a step sectioning protocol with levels of 250 microm or 200 microm would be adequate. CONCLUSIONS: SLNs are the most likely sites of nodal metastasis. Macrometastases are of recognised prognostic relevance so that all should be identified, preferably correctly as macrometastases; an assessment of levels 1 mm apart appears satisfactory and sufficient for this aim. SLNs also offer an ideal method for the study of the significance of micrometastases; for this, step sections separated by 200 or 250 microm are a good choice. PMID- 15113853 TI - HPV and histological status of pelvic lymph node metastases in cervical cancer: a prospective study. AB - AIMS: The purpose of this prospective study was to describe the incidence and distribution pattern of human papillomavirus (HPV) DNA in intraoperative dissected lymph nodes and to relate this to the pathological confirmation of metastasis. METHODS: Samples of primary cervical cancer lesions and dissected lymph nodes were obtained from women undergoing surgical treatment. The presence of HPV DNA was detected by the polymerase chain reaction. RESULTS: Tissue from 79 tumours and 365 lymph nodes was analysed. Metastasis to the lymph nodes was found in 19 cases. Metastasis correlated with the volume of the primary lesion, the depth of cervical and vaginal invasion, and with invasion of the corpus. HPV DNA was found in 60 of the primary lesions and 31 of the lymph nodes. The presence of HPV DNA in the lymph nodes correlated with the volume of the primary lesion and vaginal invasion. CONCLUSIONS: The incidence of HPV DNA in lymph nodes is twice as high as that of histopathologically confirmed metastases. The risk of the presence of HPV DNA and histopathologically confirmed metastases in lymph nodes is related to certain features of the primary tumour. PMID- 15113854 TI - Augmented pulmonary IL-4 and IL-13 receptor subunit expression in idiopathic interstitial pneumonia. AB - BACKGROUND: Some idiopathic interstitial pneumonias (IIPs) are characterised by fibroproliferation and deposition of extracellular matrix. Because efficacious treatment options are limited, research has been directed towards understanding the cytokine networks that may affect fibroblast activation and, hence, the progression of certain IIPs. AIMS: To examine the expression of interleukin 4 (IL 4), IL-13, and their corresponding receptor subunits in the various forms of IIP and normal patient groups. METHODS: Molecular and immunohistochemical analysis of IL-4, interferon gamma (IFNgamma), IL-13, IL-4 receptor (IL-R), and IL-13 receptor subunits in surgical lung biopsies (SLBs) from 39 patients (21 usual interstitial pneumonia (UIP), six non-specific interstitial pneumonia (NSIP), eight respiratory bronchiolitic interstitial lung disease (RBILD), and five normal controls). RESULTS: Molecular analysis demonstrated that IL-13Ralpha2, IL 13Ralpha1, and IL-4Ralpha were present in a greater proportion of upper and lower lobe biopsies from patients with UIP than patients with NSIP and RBILD. Immunohistochemical analysis of patients with UIP, NSIP, and RBILD revealed interstitial staining for all three receptor subunits, whereas such staining was only seen in mononuclear cells present in normal SLBs. Fibroblastic foci in patients with UIP strongly stained for IL-4Ralpha and IL-13Ralpha2. Localised expression of IL-4Ralpha was also seen in SLBs from patients with NSIP but not in other groups. CONCLUSION: Some histological subtypes of IIP are associated with increased pulmonary expression of receptor subunits responsive to IL-4 and IL-13. These findings may be of particular importance in understanding the pathogenesis of IIP and, more importantly, may provide important novel therapeutic targets. PMID- 15113855 TI - Epstein-Barr virus in gastric carcinomas and gastric stump carcinomas: a late event in gastric carcinogenesis. AB - BACKGROUND: To determine at what stage during gastric carcinogenesis Epstein-Barr virus (EBV) enters the gastric epithelial cells, the presence of EBV was investigated in two pathogenetically related but distinct forms of adenocarcinoma of the stomach-gastric carcinoma of the intact stomach (GCIS) and gastric stump carcinoma (GSC)-and their presumed precursor lesions. PATIENTS AND METHODS: Eleven patients with EBV positive GCIS and eight patients with EBV positive GSC, demonstrated by the highly sensitive EBV encoded RNA 1/2 (EBER1/2) RNA in situ hybridisation (RISH) technique, were studied. Paraffin wax embedded tissue available from preoperative gastric biopsies and tumour adjacent tissue from the resection specimens containing normal gastric mucosa, inflamed gastric mucosa, and preneoplastic lesions (intestinal metaplasia and dysplasia) was investigated by EBER1/2 RISH, in addition to EBV nuclear antigen 1 (EBNA-1) and latent membrane protein 1 (LMP-1) immunohistochemistry (IHC). RESULTS: In both GCIS and GSC and their precursor lesions EBER1/2 transcripts were restricted to the carcinoma cells. In addition, positivity of EBNA-1 IHC was also restricted to the tumour cells. IHC for LMP-1 was negative in all cases tested. CONCLUSIONS: The absence of EBER1/2 transcripts in preneoplastic gastric lesions (intestinal metaplasia and dysplasia) and their presence in two distinct types of gastric carcinoma strongly suggest that EBV can only infect neoplastic gastric cells and thus is a late event in gastric carcinogenesis. PMID- 15113856 TI - p73alpha is a candidate effector in the p53 independent apoptosis pathway of cisplatin damaged primary murine colonocytes. AB - AIMS: Colonocytes were derived from wild-type (wt) and p53 deficient mice to investigate p53 dependent and independent death pathways after cisplatin treatment, and the role of p53 in growth regulation of primary, untransformed epithelial cells. METHODS: Wt and p53 null colonocytes were exposed to cisplatin and DNA synthesis, apoptosis, and p53, p21, and p73 expression were investigated after six, 12, and 24 hours. Major p73 isoforms were identified by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Cisplatin treated wt cells exhibited cell cycle arrest, whereas p53 null cells continued to synthesise DNA, although both cell types died. Apoptosis was significantly higher in cisplatin treated wt and p53 null colonocytes than in controls at all timepoints, although apoptosis was lower in cisplatin treated p53 null colonocytes than in wt cells. p53 expression was upregulated in cisplatin treated wt colonocytes. p21 expression was high and remained unchanged in cisplatin treated wt cells, although it was reduced in the absence of p53. p73 was investigated because it could account for p53 independent p21 expression and p53 independent death. RT PCR detected full length p73alpha. p73 transcript levels remained unchanged, whereas p73 protein accumulated in the nucleus of cisplatin treated cells, irrespective of genotype. CONCLUSIONS: p53 is essential for cell cycle arrest, but not apoptosis in primary murine colonocytes. Apoptosis is reduced in cisplatin treated p53 null cells. Nuclear accumulation of endogenous p73 after cisplatin treatment suggests a proapoptotic role for p73alpha in the absence of p53 and collaboration with p53 in wt colonocytes. PMID- 15113857 TI - In vivo and in vitro studies on Anaplasma phagocytophilum infection of the myeloid cells of a patient with chronic myelogenous leukaemia and human granulocytic ehrlichiosis. AB - AIMS: The occurrence of human granulocytic ehrlichiosis (HGE) in a patient with chronic myelogenous leukaemia (CML) provided an opportunity to study whether Anaplasma phagocytophilum, the aetiological agent of HGE, infects mature or immature cells, both in vivo and in vitro. METHODS: Diagnosis of HGE was confirmed by culture, polymerase chain reaction (PCR), detection of intragranulocytic inclusions, and serology. The infection rates of different myelogenous stages of granulocytic differentiation were determined by microscopy. Anaplasma phagocytophilum infection of the bone marrow was analysed by PCR, culture, and microscopy. In addition, the in vitro growth of A phagocytophilum in the patient's granulocytes and in HL-60 cells (a promyelocytic leukaemia cell line) was compared. RESULTS: Pretreatment blood smears showed that mature granulocytic cells had a higher infection rate with A phagocytophilum than did immature cells. In the original inoculation of the patient's cells into HL-60 cells to isolate A phagocytophilum, the bacterium grew faster in the patient's leukaemic cells than in HL-60 cells. Anaplasma phagocytophilum inclusions were rarely seen in bone marrow granulocytes and PCR was negative. In vitro, two A phagocytophilum isolates grew faster in the patient's granulocytes than in HL-60 cells. CONCLUSIONS: The superior growth in CML cells compared with HL-60 cells suggests that A phagocytophilum preferentially infects mature granulocytes. The higher infection rate of the patient's mature versus immature granulocytes before treatment and the minimal level of infection of the patient's bone marrow support this. It is possible that the primary site of infection in HGE is the peripheral mature granulocytic population. PMID- 15113858 TI - Expression of vascular endothelial growth factor, hypoxia inducible factor 1alpha, and carbonic anhydrase IX in human tumours. AB - AIMS: To measure vascular endothelial growth factor (VEGF-A) mRNA in a large, diverse cohort of tumours and to investigate whether VEGF-A expression is associated with markers of hypoxia, including hypoxia inducible factor 1alpha (HIF-1alpha) and carbonic anhydrase IX (CA9). METHODS: The expression of VEGF-A and CA9 was assessed in 5067 fresh frozen human tissue samples and 238 cell lines by DNA microarray analysis. In addition, tissue microarrays were constructed from 388 malignancies to investigate the expression of VEGF-A and HIF-1alpha by in situ hybridisation and immunohistochemistry, respectively. RESULTS: VEGF-A was significantly upregulated in primary malignancies of the breast, cervix, colon and rectum, oesophagus, head and neck, kidney, ovary, skin, urinary system, and white blood cells by DNA microarray analysis. However, VEGF-A expression only correlated with CA9 expression in renal tissues. In the tissue microarrays, HIF 1alpha positive cores showed a significant increase in VEGF-A expression in lung, ovary, soft tissue, and thyroid malignancies. CONCLUSIONS: The expression of VEGF A is upregulated in a large proportion of human malignancies, and may be associated with markers of hypoxia. VEGF-A expression can be induced in the absence of hypoxia and hypoxia does not always provoke VEGF-A upregulation in tumours. PMID- 15113859 TI - Evidence that protease activated receptor 2 expression is enhanced in human coronary atherosclerotic lesions. AB - AIM: To investigate protease activated receptor 2 (PAR-2) expression in human coronary atherosclerotic lesions because PAR-2 is involved in the modulation of inflammatory events and vascular function. METHODS: An immunohistochemical analysis was performed on serial arterial sections, using the following antibodies: MDA2, a murine monoclonal antibody against malondialdehyde lysine epitopes of oxidised low density lipoprotein (oxLDL); HAM-56, a monoclonal antibody against human macrophages/foam cells; B5, a rabbit polyclonal antibody against PAR-2; and SAM11, a mouse monoclonal antibody against human PAR-2. Sections containing at least one lesion showing substantial immunostaining were counted as positive, and results were expressed as per cent of all sections of the same artery. RESULTS: PAR-2 expression was enhanced in human coronary atherosclerotic lesions. This phenomenon correlated with an increase in oxLDL epitopes in the coronary artery. CONCLUSION: This study shows for the first time that PAR-2 expression is enhanced in human coronary atherosclerotic lesions, and suggests that PAR-2 dependent cellular trafficking may be one of the regulatory signalling responses to vascular injury. Further pharmacological studies will establish whether modulation (and in which direction) of PAR-2 represents a possible therapeutic target for controlling the vascular response to injury. PMID- 15113860 TI - The prevalence and spectrum of alpha and beta thalassaemia in Guangdong Province: implications for the future health burden and population screening. AB - AIM: Thalassaemia is a good candidate disease for control by preventive genetic programmes in developing countries. Accurate population frequency data are needed for planning the control of thalassaemia in the high risk Guangdong Province of southern China. METHODS: In total, 13397 consecutive samples from five geographical areas of Guangdong Province were analysed for both haematological and molecular parameters. RESULTS: There was a high prevalence of carriers of alpha thalassaemia (8.53%), beta thalassaemia (2.54%), and both alpha and beta thalassaemia (0.26%). Overall, 11.07% of the population in this area were heterozygous carriers of alpha and beta thalassaemia. The mutation spectrum of alpha and beta thalassaemia and its constitution were fully described in this area. This study reports the true prevalence of silent alpha thalassaemia in the southern China population for the first time. In addition, two novel mutations that give rise to alpha thalassaemia, one deletion resulting in beta thalassaemia, and a rare deletion (--(THAI) allele) previously unreported in mainland China were detected. The frequency of the most common mutation, the Southeast Asian type of deletion (--(SEA), accounting for 48.54% of all alpha thalassaemias) was similar to the total of two alpha(+) thalassaemia deletions ( alpha(3.7) and -alpha(4.2), accounting for 47.49% of alpha thalassaemia). CONCLUSION: Both alpha and beta thalassaemia are widely distributed in Guangdong Province of China. The knowledge gained in this study will enable the projected number of pregnancies at risk to be estimated and a screening strategy for control of thalassaemia to be designed in this area. PMID- 15113861 TI - The favourable prognostic value of oestrogen receptor beta immunohistochemical expression in breast cancer. AB - AIMS: Oestrogen receptor beta (ERbeta) is present in breast tumours, although its prognostic and pathophysiological roles remain to be established. METHODS: Standard immunohistochemistry with a specific monoclonal antibody was performed on paraffin wax embedded sections; 10% of strongly immunostained carcinoma cells was used as the cutoff point to classify tumours as ERbeta positive. Statistical correlations were sought with clinicopathological variables (including hormone receptor status) and disease free (DFS) and overall survival (OS) in a well documented series of 181 invasive breast carcinomas. Cell proliferation was assessed immunohistochemically by topoisomerase IIa (TopoIIa) index; p53 protein accumulation and c-erbB-2 oncoprotein expression were also taken into account. RESULTS: ERbeta immunoreactivity was detected in most specimens (71.2%); it was positively linked to ERalpha immunoreactivity and increased TopoIIalpha index, and inversely to c-erbB-2 overexpression. There were no correlations with p53 immunostaining or other clinicopathological parameters. A significant favourable impact of ERbeta immunopositivity emerged with regard to DFS and OS in both univariate and multivariate analysis; ERbeta immunopositivity retained its favourable significance with regard to DFS in the subgroups of stage I and II patients when they were examined separately. Progesterone receptor expression also had an independent favourable influence on survival, albeit with less significance. In contrast, survival was not significantly influenced by ERalpha status. CONCLUSIONS: Because of the positive association between ERbeta immunoreactivity and TopoIIalpha expression, the presence of ERbeta in breast cancer cells could be considered an indication of increased proliferation. Nevertheless, ERbeta immunoreactivity emerges as a valuable, independent indicator of favourable prognosis. PMID- 15113862 TI - Human herpesvirus 8 (HHV8) sequence variations in HHV8 related tumours in Okinawa, a subtropical island in southern Japan. AB - BACKGROUND: Although rare in mainland Japan, classic Kaposi's sarcoma (KS) is frequently reported in Okinawa, a subtropical island in southern Japan. Human herpesvirus 8 (HHV8) has been identified in the tumours and geographical differences occur. AIM: To sequence HHV8 in classic and AIDS associated KS in Okinawa. MATERIALS/METHODS: Eight classic KS cases, one AIDS associated KS, five granuloma pyogenicum cases, two inflammatory pseudotumours, two Castleman's disease cases, one angiosarcoma, and one primary effusion lymphoma (PEL) were studied. As a control, HHV8 positive cultured PEL cells (TY-1) were used. The presence of HHV8 sequences was evaluated by PCR and in situ hybridisation. PCR products were sequenced. RESULTS: There were no histological differences among KS resulting from the different virus genotypes. HHV8 was detected in all cases of KS, in one PEL, and one granuloma pyogenicum. Eight classic KS cases and one granuloma pyogenicum were infected with HHV8 genotype II/C (K1 region) or subtype C (ORF26 region), which had a five amino acid deletion at K1 VR2 region. An AIDS associated KS and a PEL were infected with type I/A virus. CONCLUSION: In Okinawa, classic KS cases and one granuloma pyogenicum case were infected with HHV8 genotype II/C, also classified as subtype C. AIDS associated KS and PEL were infected with a different HHV8 (genotype I/A), similar to that found in the USA. In Okinawa, HHV8 infection is more than four times higher than in mainland Japan, resulting in many cases of KS because of HHV8 genotype II/C infection. PMID- 15113863 TI - A patient with a VEGF and endostatin producing gastrointestinal autonomic nerve tumour. AB - Tumour associated neovascularisation has been characterised as chaotic and insufficient. This report details the results of the analysis of angiogenic factors in tumour cyst fluid, pleural fluid, and blood from a patient with a gastrointestinal autonomic nerve tumour. The tumour produced vascular endothelial growth factor and endostatin in large quantities, which may explain the dysfunctional angiogenesis and tendency to bleeding seen in this tumour type. PMID- 15113864 TI - Unusual splenic sinusoidal iron overload in sickle cell/haemoglobin D-Punjab disease. AB - Sickle cell/haemoglobin D-Punjab disease is a disorder with similar clinical features to sickle cell anaemia. This report describes the case of an 11 year old boy with this disease who was treated with regular transfusions from infancy. He underwent splenectomy at the age of 10 years for hypersplenism. Histology of the spleen revealed a striking pattern of heavy sinusoidal endothelial iron loading, with only moderate uptake by macrophages. Possible explanations for this unusual distribution of iron include phagocytosis of sickled erythrocytes by sinusoidal endothelial cells or direct endothelial iron uptake via transferrin receptors. Transfusion programmes ameliorate the symptoms of sickle cell disease but the dangers of iron overload should always be remembered. PMID- 15113865 TI - Sudden death of a patient with primary hypereosinophilia, colon tumours, and pulmonary emboli. AB - A 33 year old man was admitted to hospital six days after the onset of abdominal pain. There was hypereosinophilia, but the cause could not be identified (primary hypereosinophilia). The hypereosinophilia, high C reactive protein concentration, and gastrointestinal symptoms were alleviated by corticosteroid treatment. Unexpectedly, after this apparent recovery, he was found dead on the 27th day after admission. Necropsy disclosed two solid tumours primarily composed of eosinophils in the ascending and transverse colon. The cause of the sudden death was pulmonary artery emboli, derived from a thrombus in the left iliac vein. PMID- 15113866 TI - Acute myeloid leukaemia presenting as cholestatic hepatitis. AB - A 40 year old man presented with abdominal pain, jaundice, weight loss, and hepatosplenomegaly. Liver function tests revealed cholestatic jaundice and a computed tomography scan showed an enlarged liver, with a normal biliary tree. Liver biopsy showed diffuse infiltration by neutrophils, monocytoid cells, and blasts. Peripheral blood film and bone marrow were consistent with acute myeloid leukaemia. After treatment with chemotherapy using an acute myeloid leukaemia protocol (UK Medical Research Council AML-12), there was complete resolution of jaundice and the patient went into complete molecular remission. PMID- 15113867 TI - Solitary fibrous tumour of the falciform ligament containing multiple foci of malignant transformation. AB - Solitary fibrous tumour is an uncommon soft tissue tumour initially described in the pleural cavity. In the past decade, it has been described in various extrapleural sites, including the abdomen. Traditionally regarded as benign, cases of histologically or clinically malignant solitary fibrous tumours are rare. This report describes such a case, although the patient is still disease free five years after excisional surgery, without adjuvant treatment. The acute clinical presentation may be related to malignant transformation occurring in a long standing solitary fibrous tumour. PMID- 15113868 TI - Urinary catecholamines and metabolites in the immediate postoperative period following major surgery. AB - BACKGROUND: Induction of anaesthesia can precipitate catecholamine release from an undiscovered pheochromocytoma and induce a hypertensive crisis. However, it is assumed that catecholamine and metabolite values resulting from the effects of surgery per se in the early postoperative period would overlap with the values generated by a tumour, and it is not known how soon after biochemical investigations can be carried out. AIM: To study patterns of urinary catecholamine excretion and the feasibility of biochemical screening for phaeochromocytomas in the immediate postoperative period in otherwise healthy subjects undergoing a single type of major surgical procedure. METHODS: Catecholamines and metabolites were measured for each mole of creatinine in single voided urine on one preoperative and four postoperative days in five subjects who underwent elective coronary artery bypass graft surgery with an uncomplicated postoperative course. Reference ranges were established from 33 healthy normotensive volunteers. RESULTS: Excretion of adrenaline, noradrenaline, dopamine, vanillylmandelic acid, and metadrenaline was within normal limits. Normetadrenaline excretion was mildly raised in four patients, but did not exceed 1.5 times the upper reference limit, and returned to normality by the fourth postoperative day. CONCLUSION: It is feasible to perform simple urinary screening for possible phaeochromocytoma in the immediate postoperative period. PMID- 15113869 TI - Are renal microgranulomas related to inflammatory tubular destruction? AB - Two cases of renal microgranuloma formation are reported, one in a patient with known Crohn's disease and another in a case of acute renal allograft rejection. In both cases, the microgranulomas arose as a result of inflammatory tubular destruction, in a manner analogous to that seen in patients with ulcerative colitis arising adjacent to ruptured epithelial crypts in the large intestine. Microgranulomas may occur at multiple anatomical sites in Crohn's disease, although renal microgranulomas are very rare. Non-specific inflammatory tubular destruction should be considered as a cause of renal microgranuloma formation, in addition to systemic granulomatous diseases, such as tuberculosis, sarcoid, or Crohn's disease, when granulomas are seen in the presence of inflammatory tubular destruction in renal biopsies. PMID- 15113870 TI - COX-2 dependent PGE(2) downregulates alpha(v) integrin expression via the EP(3) receptor in cultured mesangial cells. AB - BACKGROUND: In experimental glomerulonephritis, inhibition of cyclooxygenase 2 (COX-2) enhances the renocortical expression of pathogenic alpha(v) integrins. AIMS: To study whether this effect is mediated by prostaglandin E(2) (PGE(2)) acting through its EP(3) receptor in cultured rat mesangial cells (MCs). METHODS: MCs were incubated with lipopolysaccharide (LPS), celecoxib, PGE(2), or the selective EP(3) agonist, MB28767. The expression of COX-2, EP(3), and alpha(v) integrin mRNA was measured by reverse transcriptase polymerase chain reaction. RESULTS: LPS upregulated COX-2 expression 2.8-fold and alpha(v) integrin expression twofold. The COX-2 inhibitor celecoxib increased alpha(v) integrin mRNA expression twofold. Both exogenous PGE(2) and the specific EP(3) receptor agonist, MB28767, reduced constitutive alpha(v) integrin mRNA expression to half normal values. COX-2 dependent PGE(2) suppressed the expression of alpha(v) integrin mRNA mediated by the EP(3) receptor in MCs. CONCLUSIONS: These results suggest that COX-2 suppresses the expression of alpha(v) integrins by an increased production of PGE(2) activating its EP(3) receptor in glomerulonephritis. PMID- 15113871 TI - Focal nodular hyperplasia with concomitant hepatocellular carcinoma: a case report and clonal analysis. AB - This report describes a hepatocellular carcinoma (HCC) with concomitant focal nodular hyperplasia (FNH) in a 56 year old Chinese man. There were two well circumscribed tumours measuring 3 x 2.5 x 2 cm and 2 x 1.5 x 1.5 cm. The larger mass was grey and soft with a small area of bleeding and necrosis and an intact capsule. The smaller mass was yellow and had no capsule. Clonal analysis was carried out to clarify the relation between the HCC and the adjacent FNH. The clonal analysis was based on the methylation pattern of the polymorphic X chromosome linked androgen receptor gene (HUMARA). In FNH, after HpaII digestion, the allelic bands showed two well defined peaks. The intensity of the two peaks in the DNA from cirrhotic tissue did not differ significantly, consistent with a random pattern of X chromosome inactivation. However, in HCC, after HpaII digestion, the allelic bands differed significantly in intensity. Therefore, there was a typical polyclonal pattern of inactivation in FNH but the HCC was interpreted as being monoclonal. PMID- 15113872 TI - Are growth phases exclusive to cutaneous melanoma? PMID- 15113873 TI - Assessment of reliability of microarray data and estimation of signal thresholds using mixture modeling. AB - DNA microarray is an important tool for the study of gene activities but the resultant data consisting of thousands of points are error-prone. A serious limitation in microarray analysis is the unreliability of the data generated from low signal intensities. Such data may produce erroneous gene expression ratios and cause unnecessary validation or post-analysis follow-up tasks. In this study, we describe an approach based on normal mixture modeling for determining optimal signal intensity thresholds to identify reliable measurements of the microarray elements and subsequently eliminate false expression ratios. We used univariate and bivariate mixture modeling to segregate the microarray data into two classes, low signal intensity and reliable signal intensity populations, and applied Bayesian decision theory to find the optimal signal thresholds. The bivariate analysis approach was found to be more accurate than the univariate approach; both approaches were superior to a conventional method when validated against a reference set of biological data that consisted of true and false gene expression data. Elimination of unreliable signal intensities in microarray data should contribute to the quality of microarray data including reproducibility and reliability of gene expression ratios. PMID- 15113874 TI - Changes of the secondary structure of the 5' end of the Sindbis virus genome inhibit virus growth in mosquito cells and lead to accumulation of adaptive mutations. AB - Both the 5' end of the Sindbis virus (SIN) genome and its complement in the 3' end of the minus-strand RNA synthesized during virus replication serve as parts of the promoters recognized by the enzymes that comprise the replication complex (RdRp). In addition to the 5' untranslated region (UTR), which was shown to be critical for the initiation of replication, another 5' sequence element, the 51 nucleotide (nt) conserved sequence element (CSE), was postulated to be important for virus replication. It is located in the nsP1-encoding sequence and is highly conserved among all members of the Alphavirus genus. Studies with viruses containing clustered mutations in this sequence demonstrated that this RNA element is dispensable for SIN replication in cells of vertebrate origin, but its integrity can enhance the replication of SIN-specific RNAs. However, we showed that the same mutations had a deleterious effect on virus replication in mosquito cells. SIN with a mutated 51-nt CSE rapidly accumulated adaptive mutations in the nonstructural proteins nsP2 and nsP3 and the 5' UTR. These mutations functioned synergistically in a cell-specific manner and had a stimulatory effect only on the replication of viruses with a mutated 51-nt CSE. Taken together, the results suggest the complex nature of interactions between nsP2, nsP3, the 5' UTR, and host-specific protein factors binding to the 51-nt CSE and involved in RdRp formation. The data also demonstrate an outstanding potential of alphaviruses for adaptation. Within one passage, SIN can adapt to replication in cells of a vertebrate or invertebrate origin. PMID- 15113876 TI - Loss of N-linked glycosylation from the hemagglutinin-neuraminidase protein alters virulence of Newcastle disease virus. AB - The hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) is an important determinant of its virulence. We investigated the role of each of the four functional N-linked glycosylation sites (G1 to G4) of the HN glycoprotein of NDV on its pathogenicity. The N-linked glycosylation sites G1 to G4 at residues 119, 341, 433, and 481, respectively, of a moderately pathogenic NDV strain Beaudette C (BC) were eliminated individually by site-directed mutagenesis on a full-length cDNA clone of BC. A double mutant (G12) was also created by eliminating the first and second glycosylation sites at residues 119 and 341, respectively. Infectious virus was recovered from each of the cDNA clones of the HN glycoprotein mutants, employing a reverse genetics technique. There was a greater delay in the replication of G4 and G12 mutant viruses than in the parental virus. Loss of glycosylation does not affect the receptor recognition by HN glycoprotein of NDV. The neuraminidase activity of G4 and G12 mutant viruses and the fusogenicity of the G4 mutant virus were significantly lower than those of the parental virus. The fusogenicity of the double mutant virus (G12) was significantly higher than that of the parental virus. Cell surface expression of the G4 virus HN was significantly lower than that of the parental virus. The antigenic reactivities of the mutants to a panel of monoclonal antibodies against the HN protein indicated that removal of glycosylation from the HN protein increased (G1, G3, and G12) or decreased (G2 and G4) the formation of antigenic sites, depending on their location. In standard tests to assess virulence in chickens, all of the glycosylation mutants were less virulent than the parental BC virus, but the G4 and G12 mutants were the least virulent. PMID- 15113877 TI - The plant virus Tomato Spotted Wilt Tospovirus activates the immune system of its main insect vector, Frankliniella occidentalis. AB - Tospoviruses have the ability to infect plants and their insect vectors. Tomato spotted wilt virus (TSWV), the type species in the Tospovirus genus, infects its most important insect vector, Frankliniella occidentalis, the western flower thrips (WFT). However, no detrimental effects on the life cycle or cytopathological changes have been reported in the WFT after TSWV infection, and relatively few viral particles can be observed even several days after infection. We hypothesized that TSWV infection triggers an immune response in the WFT. Using subtractive cDNA libraries to probe WFT DNA macroarrays, we found that the WFT's immune system is activated by TSWV infection. The activated genes included (i) those encoding antimicrobial peptides, such as defensin and cecropin; (ii) genes involved in pathogen recognition, such as those encoding lectins; (iii) those encoding receptors that activate the innate immune response, such as Toll-3; and (iv) those encoding members of signal transduction pathways activated by Toll like receptors, such as JNK kinase. Transcriptional upregulation of these genes after TSWV infection was confirmed by Northern analysis, and the kinetics of the immune response was measured over time. Several of the detected genes were activated at the same time that viral replication was first detected by reverse transcription-PCR. To our knowledge, this is the first report of the activation of an insect vector immune response by a plant virus. The results may lead to a better understanding of insects' immune responses against viruses and may help in the future development of novel control strategies against plant viruses, as well as human and animal viruses transmitted by insect vectors. PMID- 15113878 TI - The BRRF1 early gene of Epstein-Barr virus encodes a transcription factor that enhances induction of lytic infection by BRLF1. AB - The switch from the latent to the lytic form of Epstein-Barr virus (EBV) infection is mediated by expression of the viral immediate-early (IE) proteins, BZLF1 (Z) and BRLF1 (R). An EBV early protein, BRRF1 (Na), is encoded by the opposite strand of the BRLF1 intron, but the function of this nuclear protein in the viral life cycle is unknown. Here we demonstrate that Na enhances the R mediated induction of lytic EBV infection in 293 cells latently infected with a recombinant EBV (R-KO) defective for the expression of both R and Na. Na also enhances R-induced lytic infections in a gastric carcinoma line (AGS) carrying the R-KO virus, although it has no effect in a Burkitt lymphoma line (BL-30) stably infected with the same mutant virus. We show that Na is a transcription factor that increases the ability of R to activate Z expression from the R-KO viral genome in 293 cells and that Na by itself activates the Z promoter (Zp) in EBV-negative cells. Na activation of Zp requires a CRE motif (ZII), and a consensus CRE motif is sufficient to transfer Na responsiveness to the heterologous E1b promoter. Furthermore, we show that Na enhances the transactivator function of a Gal4-c-Jun fusion protein but does not increase the transactivator function of other transcription factors (including ATF-1, ATF-2, and CREB) known to bind CRE motifs. Na expression in cells results in increased levels of a hyperphosphorylated form of c-Jun, suggesting a mechanism by which Na activates c-Jun. Our results indicate that Na is a transcription factor that activates the EBV Zp IE promoter through its effects on c-Jun and suggest that Na cooperates with BRLF1 to induce the lytic form of EBV infection in certain cell types. PMID- 15113879 TI - A sequence-independent strategy for detection and cloning of circular DNA virus genomes by using multiply primed rolling-circle amplification. AB - The discovery of novel viruses has often been accomplished by using hybridization based methods that necessitate the availability of a previously characterized virus genome probe or knowledge of the viral nucleotide sequence to construct consensus or degenerate PCR primers. In their natural replication cycle, certain viruses employ a rolling-circle mechanism to propagate their circular genomes, and multiply primed rolling-circle amplification (RCA) with phi29 DNA polymerase has recently been applied in the amplification of circular plasmid vectors used in cloning. We employed an isothermal RCA protocol that uses random hexamer primers to amplify the complete genomes of papillomaviruses without the need for prior knowledge of their DNA sequences. We optimized this RCA technique with extracted human papillomavirus type 16 (HPV-16) DNA from W12 cells, using a real time quantitative PCR assay to determine amplification efficiency, and obtained a 2.4 x 10(4)-fold increase in HPV-16 DNA concentration. We were able to clone the complete HPV-16 genome from this multiply primed RCA product. The optimized protocol was subsequently applied to a bovine fibropapillomatous wart tissue sample. Whereas no papillomavirus DNA could be detected by restriction enzyme digestion of the original sample, multiply primed RCA enabled us to obtain a sufficient amount of papillomavirus DNA for restriction enzyme analysis, cloning, and subsequent sequencing of a novel variant of bovine papillomavirus type 1. The multiply primed RCA method allows the discovery of previously unknown papillomaviruses, and possibly also other circular DNA viruses, without a priori sequence information. PMID- 15113880 TI - Treatment of transmissible spongiform encephalopathy by intraventricular drug infusion in animal models. AB - The therapeutic efficacy of direct drug infusion into the brain, the target organ of transmissible spongiform encephalopathies, was assessed in transgenic mice intracerebrally infected with 263K scrapie agent. Pentosan polysulfate (PPS) gave the most dramatic prolongation of the incubation period, and amphotericin B had intermediate effects, but antimalarial drugs such as quinacrine gave no significant prolongation. Treatment with the highest dose of PPS at an early or late stage of the infection prolonged the incubation time by 2.4 or 1.7 times that of the control mice, respectively. PPS infusion decreased not only abnormal prion protein deposition but also neurodegenerative changes and infectivity. These alterations were observed within the brain hemisphere fitted with an intraventricular infusion cannula but not within the contralateral hemisphere, even at the terminal disease stage long after the infusion had ended. Therapeutic effects of PPS were also demonstrated in mice infected with either RML agent or Fukuoka-1 agent. However, at doses higher than that providing the maximal effects, intraventricular PPS infusion caused adverse effects such as hematoma formation in the experimental animals. These findings indicate that intraventricular PPS infusion might be useful for the treatment of transmissible spongiform encephalopathies in humans, providing that the therapeutic dosage is carefully evaluated. PMID- 15113881 TI - Use of gHgL for attachment of Epstein-Barr virus to epithelial cells compromises infection. AB - Epstein-Barr virus (EBV) is a lymphotropic herpesvirus. However, access to B lymphocytes during primary infection may be facilitated by replication in mucosal epithelial cells. Attachment and penetration of EBV into these two cell types are fundamentally different. Both the distribution of receptors and the cellular origin of the virus impact the efficiency of infection. Epithelial cells potentially offer a wide range of receptors with which virus can interact. We report here on analyses of epithelial cells expressing different combinations of receptors. We find that the stoichiometry of the virus glycoprotein complex that includes gHgL and gp42 affects the use of gHgL not just for entry into epithelial cells but also for attachment. Penetration can be mediated efficiently with either a coreceptor for gp42 or gHgL, but the use of gHgL for attachment as well as penetration greatly compromises its ability to mediate entry. PMID- 15113882 TI - RhoA-derived peptide dimers share mechanistic properties with other polyanionic inhibitors of respiratory syncytial virus (RSV), including disruption of viral attachment and dependence on RSV G. AB - Large polyanionic molecules, such as sulfated polysaccharides (including soluble heparin and dextran sulfate), synthetic polyanionic polymers, and negatively charged proteins, have been shown to broadly inhibit several enveloped viruses. We recently reported the antiviral activity of a peptide derived from amino acids 77 to 95 of a potential binding partner of respiratory syncytial virus F protein (RSV F), the GTPase RhoA. A subsequent study with a truncated peptide (amino acids 80 to 94) revealed that optimal antiviral activity required dimerization via intermolecular disulfide bonds. We report here that the net negative charge of this peptide is also a determining factor for its antiviral activity and that it, like other polyanions, inhibits virus attachment. In a flow cytometry-based binding assay, peptide 80-94, heparin, and dextran sulfate inhibited the attachment of virus to cells at 4 degrees C at the same effective concentrations at which they prevent viral infectivity. Interestingly, time-of-addition experiments revealed that peptide 80-94 and soluble heparin were also able to inhibit the infectivity of a virus that had been prebound to cells at 4 degrees C, as had previously been shown for dextran sulfate, suggesting a potential role for postattachment effects of polyanions on RSV entry. Neutralization experiments with recombinant viruses showed that the antiviral activities of peptide 80-94 and dextran sulfate were diminished in the absence of the RSV attachment glycoprotein (G). Taken together, these data indicate that the antiviral activity of RhoA-derived peptides is functionally similar to that of other polyanions, is dependent on RSV G, and does not specifically relate to a protein-protein interaction between F and RhoA. PMID- 15113883 TI - An early stage of Mason-Pfizer monkey virus budding is regulated by the hydrophobicity of the Gag matrix domain core. AB - Intracellular capsid transport and release of Mason-Pfizer monkey virus are dependent on myristylation of the Gag matrix domain (MA). A myristylated MA mutant, in which Thr41 and Thr78 are replaced with isoleucines, assembles capsids that are transported to the plasma membrane but are blocked in an early budding step. Since the nuclear magnetic resonance structure of MA showed that these Thr residues point into the hydrophobic core of the protein, it was hypothesized that the T41I/T78I mutant was defective in release of myristic acid from the more hydrophobic core. In order to further investigate whether an increase in the hydrophobicity of the MA core modulates capsid-membrane interactions and viral budding, three tyrosine residues (11, 28, and 67), oriented toward the MA core, were replaced individually or in a pair-wise combination with the more hydrophobic phenylalanine residue(s). As a control, Tyr82, oriented toward the outer surface of MA, was also replaced with phenylalanine. These Tyr-to-Phe substitutions did not alter capsid assembly compared to wild type in a capsid assembly assay. Pulse-chase, immunofluorescence, and electron microscopy studies demonstrated that single substitutions of Tyr11, Tyr28, and Tyr67 recapitulated the T41I/T78I mutant phenotype of decreased budding kinetics and accumulation of capsids at the plasma membrane. MA double mutants with a combination of these Tyr substitutions exhibited a phenotype that was even more defective in budding. In contrast, MA mutants with Tyr82 replaced by Phe resulted in a transport-defective phenotype. These results strongly support the hypothesis that myristic acid is sequestered inside MA prior to capsid-membrane interactions. PMID- 15113884 TI - Activation of adenoviral gene expression by protein IX is not required for efficient virus replication. AB - The adenovirus (Ad) protein IX (pIX) is a minor component of the Ad capsid and is in part responsible for virion stability; virions lacking pIX are heat labile and lose their infectivity if the DNA content is greater than approximately 35 kb. More recently, pIX has been identified as a transcriptional activator and, in transient-transfection assays, was shown to enhance expression from the E1A, E4, and major late Ad promoters by as much as 70-fold. In this study, we examined the role of pIX's ability to activate transcription during Ad replication. In transient-transfection assays, pIX had a minimal effect on expression from the E1A promoter, increasing expression by only 1.4-fold. We used helper-dependent Ad vectors, which had all Ad protein coding sequences deleted with the exception of E1A and which had capsids that either contained or lacked pIX, to show that pIX derived from decapsidation of the infecting virion does not influence expression of E1A. Similarly, expression of pIX from the Ad genome did not alter the expression levels of E1A. Viruses that had pIX deleted showed a threefold reduction in virus yield and expression of late genes compared to those of a similar virus which encoded pIX. This phenotype could not be rescued by growing the virus in cells which constitutively express pIX. Our results indicate that, although pIX can affect transcription from a variety of viral promoters, it does not appear to play a significant role in activation of Ad promoters during normal Ad replication. PMID- 15113885 TI - Wide variations in herpes simplex virus type 1 inoculum dose and latency associated transcript expression phenotype do not alter the establishment of latency in the rabbit eye model. AB - The latency-associated transcript (LAT) is required for efficient reactivation of herpes simplex virus type 1 from latent infection in the rabbit eye model, but LAT's mechanism of action is unknown. In addition to reactivation, the LAT region seems to correspond to multiple functions, with some LAT deletion mutants exhibiting increased virulence, increased neuronal death, and restricted establishment of latency. While a LAT promoter deletion mutant (17DeltaPst) seems to be primarily restricted in reactivation in the rabbit, subtle effects on virulence or the establishment of latency cannot be precluded at the normal high levels of virus inoculum used in the rabbit model. Since such additional LAT phenotypes may be more evident with lower doses of virus, we evaluated the influence of initial viral inoculum and LAT expression on the progression of acute infection and the establishment of latency. We have assayed both virus recovery rates and viral genome loads in rabbit corneas and trigeminal ganglia. Our results show that (i) in the corneas and trigeminal ganglia, the maximum amount of virus present during acute infection is independent of the LAT genotype and inoculum dose, although greater viral yields are obtained earlier with higher inoculum doses, and (ii) the range in numbers of latent genomes detected in the ganglia is independent of the inoculum dose and the LAT genotype and therefore no difference in establishment of latency is observed. PMID- 15113886 TI - Requirement for integrase during reverse transcription of human immunodeficiency virus type 1 and the effect of cysteine mutations of integrase on its interactions with reverse transcriptase. AB - Retroviral integrase catalyzes the essential step of integrating a double stranded DNA copy of the viral genome into a host cell chromosome. Mutational studies have revealed that integrase is involved in additional steps of viral replication, but the mechanism for the pleiotropic effect is not well characterized. Since Cys residues generally play crucial roles in protein structure and function, we introduced Cys-to-Ser substitutions at positions 56, 65, and 130 of human immunodeficiency virus type 1 (HIV-1) integrase to determine their effects on integration activity and viral replication. None of the substitutions significantly affected the enzymatic activities in vitro. When introduced into the NL4-3 molecular clone of HIV-1, mutant viruses encoding Cys mutations at positions 56 and 65 of integrase replicated similarly to the wild type virus in CD4(+)-T-cell lines, whereas the C130S-containing virus was noninfectious. The entry and postintegration steps of the viral life cycle for all mutant viruses were normal, and all had particle-associated reverse transcriptase (RT) activity. However, early reverse-transcribed DNA products were absent in the lysate of cells infected with the C130S mutant virus, indicating that the mutation abolished the ability of the virus to initiate endogenous reverse transcription. Coimmunoprecipitation using purified integrase and RT showed that the C-terminal domain of wild-type HIV-1 integrase interacted with RT. The interaction between integrase and RT was not affected in the presence of a reducing or alkylating agent, suggesting that the interaction did not involve a disulfide linkage. The C130S substitution within the core region may disrupt the protein recognition interface of the C-terminal domain and abolish its ability to interact with RT. Our results indicate that integrase plays an important role during the reverse-transcription step of the viral life cycle, possibly through physical interactions with RT. PMID- 15113887 TI - Interaction between human immunodeficiency virus type 1 reverse transcriptase and integrase proteins. AB - Reverse transcriptase (RT) and integrase (IN) are two key catalytic enzymes encoded by all retroviruses. It has been shown that a specific interaction occurs between the human immunodeficiency virus type 1 (HIV-1) RT and IN proteins (X. Wu, H. Liu, H. Xiao, J. A. Conway, E. Hehl, G. V. Kalpana, V. R. Prasad, and J. C. Kappes, J. Virol. 73:2126-2135, 1999). We have now further examined this interaction to map the binding domains and to determine the effects of interaction on enzyme function. Using recombinant purified proteins, we have found that both a HIV-1 RT heterodimer (p66/p51) and its individual subunits, p51 and p66, are able to bind to HIV-1 IN. An oligomerization-defective mutant of IN, V260E, retained the ability to bind to RT, showing that IN oligomerization may not be required for interaction. Furthermore, we report that the C-terminal domain of IN, but not the N-terminal zinc-binding domain or the catalytic core domain, was able to bind to heterodimeric RT. Deletion analysis to map the IN binding domain on RT revealed two separate IN-interacting domains: the fingers palm domain and the carboxy-terminal half of the connection subdomain. The carboxy-terminal domain of IN alone retained its interaction with both the fingers-palm and the connection-RNase H fragments of RT, but not with the half connection-RNase H fragment. This interaction was not bridged by nucleic acids, as shown by micrococcal nuclease treatment of the proteins prior to the binding reaction. The influences of IN and RT on each other's activities were investigated by performing RT processivity and IN-mediated 3' processing and joining reactions in the presence of both proteins. Our results suggest that, while IN had no influence on RT processivity, RT stimulated the IN-mediated strand transfer reaction in a dose-dependent manner up to 155-fold. Thus, a functional interaction between these two viral enzymes may occur during viral replication. PMID- 15113888 TI - Conserved cysteine-rich domain of paramyxovirus simian virus 5 V protein plays an important role in blocking apoptosis. AB - The paramyxovirus family includes many well-known human and animal pathogens as well as emerging viruses such as Hendra virus and Nipah virus. The V protein of simian virus 5 (SV5), a prototype of the paramyxoviruses, contains a cysteine rich C-terminal domain which is conserved among all paramyxovirus V proteins. The V protein can block both interferon (IFN) signaling by causing degradation of STAT1 and IFN production by blocking IRF-3 nuclear import. Previously, it was reported that recombinant SV5 lacking the C terminus of the V protein (rSV5VDeltaC) induces a severe cytopathic effect (CPE) in tissue culture whereas wild-type (wt) SV5 infection does not induce CPE. In this study, the nature of the CPE and the mechanism of the induction of CPE were investigated. Through the use of DNA fragmentation, terminal deoxynucleotidyltransferase-mediated dUTP biotin nick end labeling, and propidium iodide staining assays, it was shown that rSV5VDeltaC induced apoptosis. Expression of wt V protein prevented apoptosis induced by rSV5VDeltaC, suggesting that the V protein has an antiapoptotic function. Interestingly, rSV5VDeltaC induced apoptosis in U3A cells (a STAT1 deficient cell line) and in the presence of neutralizing antibody against IFN, suggesting that the induction of apoptosis by rSV5VDeltaC was independent of IFN and IFN-signaling pathways. Apoptosis induced by rSV5VDeltaC was blocked by a general caspase inhibitor, Z-VAD-FMK, but not by specific inhibitors against caspases 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 13, suggesting that rSV5VDeltaC induced apoptosis can occur in a caspase 12-dependent manner. Endoplasmic reticulum stress can lead to activation of caspase 12; compared to the results seen with mock and wt SV5 infection, rSV5VDeltaC infection induced ER stress, as demonstrated by increased expression levels of known ER stress indicators GRP 78, GRP 94, and GADD153. These data suggest that rSV5VDeltaC can trigger cell death by inducing ER stress. PMID- 15113889 TI - Prediction and identification of a permissive epitope insertion site in the vesicular stomatitis virus glycoprotein. AB - We developed a rational approach to identify a site in the vesicular stomatitis virus (VSV) glycoprotein (G) that is exposed on the protein surface and tolerant of foreign epitope insertion. The foreign epitope inserted was the six-amino-acid sequence ELDKWA, a sequence in a neutralizing epitope from human immunodeficiency virus type 1. This sequence was inserted into six sites within the VSV G protein (Indiana serotype). Four sites were selected based on hydrophilicity and high sequence variability identified by sequence comparison with other vesiculovirus G proteins. The site showing the highest variability was fully tolerant of the foreign peptide insertion. G protein containing the insertion at this site folded correctly, was transported normally to the cell surface, had normal membrane fusion activity, and could reconstitute fully infectious VSV. The virus was neutralized by the human 2F5 monoclonal antibody that binds the ELDKWA epitope. Additional studies showed that this site in G protein tolerated insertion of at least 16 amino acids while retaining full infectivity. The three other insertions in somewhat less variable sequences interfered with VSV G folding and transport to the cell surface. Two additional insertions were made in a conserved sequence adjacent to a glycosylation site and near the transmembrane domain. The former blocked G-protein transport, while the latter allowed transport to the cell surface but blocked membrane fusion activity of G protein. Identification of an insertion-tolerant site in VSV G could be important in future vaccine and targeting studies, and the general principle might also be useful in other systems. PMID- 15113890 TI - Differential cell killing by lymphomagenic murine leukemia viruses occurs independently of p53 activation and mitochondrial damage. AB - Upon inoculation into AKR mice, mink cell focus-forming murine leukemia virus (MCF MLV) accelerates thymic lymphoma formation. During the preleukemic phase of disease, we observed the induction of apoptosis in thymic lymphocytes. A similar induction of apoptosis was observed for cultured mink epithelial cells after MCF13 MLV infection. In this study, the relevance of viral pathogenicity to cell killing was determined by testing the susceptibility of various cell types from different species to lymphomagenic MLVs. We observed that the cytopathic effect of lymphomagenic MLVs was restricted to mink cells. Southern blot analysis of MLV infected cells revealed an accumulation of the linear form of unintegrated viral DNA, particularly in mink cells after MCF13 MLV infection. Thus, a strong correlation was observed between viral superinfection, which results in the accumulation of high levels of unintegrated viral DNA, and cell killing. Immunoblot analysis for MCF13 MLV-infected mink epithelial cells did not show a significant change in total p53 levels or its phosphorylated form at Ser-15 compared with that in mock-treated cells. Moreover, a time course analysis for mink epithelial cells infected with MCF13 MLV did not reveal mitochondrial depolarization or a significant change in Bax levels. These results demonstrate that MCF13 MLV induces apoptosis preferentially in cells in which superinfection occurs, and the mechanism involved is independent of p53 activation and mitochondrial damage. PMID- 15113891 TI - Susceptibility of cell populations to transduction by retroviral vectors. AB - Retroviral transduction efficiency is related to the multiplicity of infection and the physiological state of the target cells. It is generally not known what proportion of a cell population is susceptible to transduction. We used coinfection with two retroviral vectors containing the marker genes for green fluorescent protein and the truncated human nerve growth factor receptor. In the CD34+ cell line TF-1 or human primary CD34+ hematopoietic progenitor cells, it was found that cells transduced with one vector had a better than random chance of transduction by the other vector. A probability model was developed to estimate target cell susceptibility; susceptibility was calculated as the product of the proportions of transgene-positive cells divided by the proportion of double-positive cells. By using this relationship, it was found that susceptibility was related to the target cell type and culture conditions but not the retroviral titer or the retroviral packaging envelope protein used in this study. Cotransduction with two vectors is a relatively simple procedure that provides a means to assess the maximum transduction level possible in a given cell population. PMID- 15113892 TI - Murine gammaherpesvirus 68 lacking gp150 shows defective virion release but establishes normal latency in vivo. AB - All gammaherpesviruses encode a virion glycoprotein positionally homologous to Epstein-Barr virus gp350. These glycoproteins are thought to be involved in cell binding, but little is known of the roles they might play in the whole viral replication cycle. We have analyzed the contribution of murine gammaherpesvirus 68 (MHV-68) gp150 to viral propagation in vitro and host colonization in vivo. MHV-68 lacking gp150 was viable and showed normal binding to fibroblasts and normal single-cycle lytic replication. Its capacity to infect glycosaminoglycan (GAG)-deficient CHO-K1 cells and NS0 and RAW264.7 cells, which express only low levels of GAGs, was paradoxically increased. However, gp150-deficient MHV-68 spread poorly through fibroblast monolayers, with reduced cell-free infectivity, consistent with a deficit in virus release. Electron microscopy showed gp150 deficient virions clustered on infected-cell plasma membranes. MHV-68-infected cells showed reduced surface GAG expression, suggesting that gp150 prevented virions from rebinding to infected cells after release by making MHV-68 infection GAG dependent. Surprisingly, gp150-deficient viruses showed only a transient lag in lytic replication in vivo and established normal levels of latency. Cell-to cell virus spread and the proliferation of latently infected cells, for which gp150 was dispensable, therefore appeared to be the major route of virus propagation in an infected host. PMID- 15113893 TI - Cellular repressor inhibits human cytomegalovirus transcription from the UL127 promoter. AB - The region of the human cytomegalovirus (HCMV) genome between the UL127 promoter and the major immediate-early (MIE) enhancer is referred to as the unique region. The role of this region during a viral infection is not known. In wild-type HCMV infected permissive fibroblasts, there is no transcription from the UL127 promoter at any time during productive infection. Our investigators previously reported that the region upstream of the UL127 TATA box repressed expression from the UL127 promoter (C. A. Lundquist et al., J. Virol. 73:9039-9052, 1999). The region was reported to contain functional NF1 DNA binding sites (L. Hennighausen and B. Fleckenstein, EMBO J. 5:1367-1371, 1986). Sequence analysis of this region detected additional consensus binding sites for three transcriptional regulatory proteins, FoxA (HNF-3), suppressor of Hairy wing, and CAAT displacement protein. The cis-acting elements in the unique region prevented activation of the early UL127 promoter by the HCMV MIE proteins. In contrast, deletion of the region permitted very high activation of the UL127 promoter by the viral MIE proteins. Mutation of the NF1 sites had no effect on the basal activity of the promoter. To determine the role of the other sites in the context of the viral genome, recombinant viruses were generated in which each putative repressor site was mutated and the effect on the UL127 promoter was analyzed. Mutation of the putative Fox-like site resulted in a significant increase in expression from the viral early UL127 promoter. Insertion of wild-type Fox-like sites between the HCMV immediate-early (IE) US3 TATA box and the upstream NF-kappaB-responsive enhancer (R2) also significantly decreased gene expression, but mutated Fox-like sites did not. The wild-type Fox-like site inhibits activation of a viral IE enhancer-containing promoter. Cellular protein, which is present in uninfected or infected permissive cell nuclear extracts, binds to the wild-type Fox-like site but not to mutated sites. Reasons for repression of UL127 gene transcription during productive infection are discussed. PMID- 15113894 TI - Rapid and sensitive detection of retrovirus entry by using a novel luciferase based content-mixing assay. AB - We describe a novel assay that permits measurement of entry of murine leukemia virus and pseudotypes with greater sensitivity and more rapidly than previously possible. To achieve this, we encapsulated a sensitive reporter enzyme, luciferase, directly into fully infectious, intact viral particles. The enzyme is specifically targeted to the viral lumen, as a C-terminal fusion on the viral envelope protein. Only when the incorporated luciferase is released from the viral lumen and gains access to its substrates is light emitted and readily detected. When cells are perfused with luciferin, quantitative measurements of entry can be made in real time on live cells. Uniquely, the amount of cell-bound virus can be determined in the same assay by addition of detergent to expose the luciferase. We demonstrate that virus carrying a mutation in the fusion peptide binds normally to cells but is unable to infect them and gives no entry signal. Using this assay, we show that inhibitors of endosomal acidification inhibit signal from vesicular stomatitis virus pseudotypes but not murine leukemia virus, consistent with a pH-independent mode of entry for the latter virus. Additionally, the fusion kinetics are rapid, with a half-life of 25 min after a delay of 10 to 15 min. The future use of this assay will permit a detailed examination of the entry mechanism of viruses and provide a convenient platform to discover novel entry inhibitors. The design also permits packaging of potential therapeutic protein cargoes into functional virus particles and their specific delivery to cellular targets. PMID- 15113895 TI - Accumulation of a 3'-terminal genome fragment in Japanese encephalitis virus infected mammalian and mosquito cells. AB - Japanese encephalitis virus (JEV) contains a single positive-strand RNA genome nearly 11 kb in length and is not formally thought to generate subgenomic RNA molecules during replication. Here, we report the abundant accumulation of a 3' terminal 521- to 523-nucleotide (nt) genome fragment, representing a major portion of the 585-nt 3' untranslated region, in both mammalian (BHK-21) and mosquito (C6/36) cells infected with any of nine strains of JEV. In BHK-21 cells, the viral genome was detected as early as 24 h postinfection, the small RNA was detected as early as 28 h postinfection, and the small RNA was 0.25 to 1.5 times as abundant as the genome on a molar basis between 28 and 48 h postinfection. In C6/36 cells, the genome and small RNA were present 5 days postinfection and the small RNA was 1.25 to 5.14 times as abundant as the genome. The 3'-terminal 523 nt small RNA contains a 5'-proximal stable hairpin (nt 6 to 56) that may play a role in its formation and the conserved flavivirus 3'-cyclization motif (nt 413 to 420) and the 3'-terminal long stable hairpin structure (nt 440 to 523) that have postulated roles in genome replication. Abundant accumulation of the small RNA during viral replication in both mammalian and mosquito cells suggests that it may play a biological role, perhaps as a regulator of RNA synthesis. PMID- 15113896 TI - Unspliced Rous sarcoma virus genomic RNAs are translated and subjected to nonsense-mediated mRNA decay before packaging. AB - Retroviruses package full-length, unspliced RNAs into progeny virions as dimerized RNA genomes. They also use unspliced RNAs as mRNAs to produce the gag and pol gene products. We asked whether a single Rous sarcoma virus (RSV) RNA can be translated and subsequently packaged or whether genomic packaging requires a nontranslated population of RNAs. We addressed this issue by utilizing the translation-dependent nonsense-mediated mRNA decay (NMD) pathway. NMD is the selective destruction of mRNAs bearing premature termination codons (PTCs). The pathway has been shown to be associated with splicing in higher eukaryotes. Here, we demonstrate that both translation and the cellular factor Upf1 are required for the decay of unspliced, PTC-bearing RSV RNA by the NMD pathway. To address the relationship between RNA translation and packaging, we examined virus produced in cells cotransfected with PTC-bearing retroviral clones and wild-type viral clones. We observed that PTC-bearing transcripts are packaged into viral particles at levels three- to fivefold less than those of control RNAs. Since PTC mediated degradation requires translation, we conclude that RSV can package progeny virion particles using previously translated RNAs. PMID- 15113897 TI - Theta defensins protect cells from infection by herpes simplex virus by inhibiting viral adhesion and entry. AB - We tested the ability of 20 synthetic theta defensins to protect cells from infection by type 1 and type 2 herpes simplex viruses (HSV-1 and -2, respectively). The peptides included rhesus theta defensins (RTDs) 1 to 3, originally isolated from rhesus macaque leukocytes, and three peptides (retrocyclins 1 to 3) whose sequences were inferred from human theta-defensin (DEFT) pseudogenes. We also tested 14 retrocyclin analogues, including the retro, enantio, and retroenantio forms of retrocyclin 1. Retrocyclins 1 and 2 and RTD 3 protected cervical epithelial cells from infection by both HSV serotypes, but only retrocyclin 2 did so without causing cytotoxicity or requiring preincubation with the virus. Surface plasmon resonance studies revealed that retrocyclin 2 bound to immobilized HSV-2 glycoprotein B (gB2) with high affinity (K(d), 13.3 nM) and that it did not bind to enzymatically deglycosylated gB2. Temperature shift experiments indicated that retrocyclin 2 and human alpha defensins human neutrophil peptide 1 (HNP 1) to HNP 3 protected human cells from HSV-2 by different mechanisms. Retrocyclin 2 blocked viral attachment, and its addition during the binding or penetration phases of HSV-2 infection markedly diminished nuclear translocation of VP16 and expression of ICP4. In contrast, HNPs 1 to 3 had little effect on binding but reduced both VP16 transport and ICP4 expression if added during the postbinding (penetration) period. We recently reported that theta defensins are miniature lectins that bind gp120 of human immunodeficiency virus type 1 (HIV-1) with high affinity and inhibit the entry of R5 and X4 isolates of HIV-1. Given its small size (18 residues), minimal cytotoxicity, lack of activity against vaginal lactobacilli, and effectiveness against both HSV-2 and HIV-1, retrocyclin 2 provides an intriguing prototype for future topical microbicide development. PMID- 15113898 TI - Effect of extension of the cytoplasmic domain of human immunodeficiency type 1 virus transmembrane protein gp41 on virus replication. AB - The biological significance of the presence of a long cytoplasmic domain in the envelope (Env) transmembrane protein gp41 of human immunodeficiency virus type 1 (HIV-1) is still not fully understood. Here we examined the effects of cytoplasmic tail elongation on virus replication and characterized the role of the C-terminal cytoplasmic tail in interactions with the Gag protein. Extensions with six and nine His residues but not with fewer than six His residues were found to severely inhibit virus replication through decreased Env electrophoretic mobility and reduced Env incorporation compared to the wild-type virus. These two mutants also exhibited distinct N glycosylation and reduced cell surface expression. An extension of six other residues had no deleterious effect on infectivity, even though some mutants showed reduced Env incorporation into the virus and/or decreased cell surface expression. We further show that these elongated cytoplasmic tails in a format of the glutathione S-transferase fusion protein still interacted effectively with the Gag protein. In addition, the immediate C terminus of the cytoplasmic tail was not directly involved in interactions with Gag, but the region containing the last 13 to 43 residues from the C terminus was critical for Env-Gag interactions. Taken together, our results demonstrate that HIV-1 Env can tolerate extension at its C terminus to a certain degree without loss of virus infectivity and Env-Gag interactions. However, extended elongation in the cytoplasmic tail may impair virus infectivity, Env cell surface expression, and Env incorporation into the virus. PMID- 15113899 TI - Identification of unique hepatitis C virus quasispecies in the central nervous system and comparative analysis of internal translational efficiency of brain, liver, and serum variants. AB - Reports of cerebral dysfunction in chronic hepatitis C virus (HCV) infection have led to the suggestion that HCV may infect the central nervous system (CNS). We used reverse transcription-PCR, cloning, and sequencing to define quasispecies for the HCV internal ribosomal entry site (IRES) and hypervariable region 1 (HVR1) in autopsy-derived brain, liver, lymph node, and serum samples. There was evidence of tissue compartmentalization of sequences in the brain in two patients, with between 24 and 55% of brain-derived IRES sequences absent from the serum, and significant phylogenetic and phenetic clustering of the brain and lymph node HVR1 sequences. The IRES initiates cap-independent translation of the viral polyprotein. Two unique brain-derived IRES mutations (C(204)-->A and G(243) ->A), which have previously been associated with lymphoid replication and altered translational efficiency in cell culture, were found in one patient. We used a dicistronic reporter vector to test whether brain-derived variants showed altered IRES-mediated translational efficiency, which might favor CNS infection. The translational efficiencies of the brain-derived IRES sequences were generally reduced compared to those of the master serum and liver sequences in rabbit reticulocyte cell lysates and two human cell lines, HuH7 (liver) and CHME3 (microglial). The C(204)-->A and G(243)-->A mutations showed preserved translational efficiency in HuH7 cells but reduced efficiency in CHME3 cells. Our data provide evidence that the CNS is a site of HCV replication, consistent with the recent demonstration of negative-strand HCV RNA in brain, and suggest that IRES polymorphisms may be important as a viral strategy of reduced translation to favor latency in the CNS. PMID- 15113900 TI - Migration of antigen-specific T cells away from CXCR4-binding human immunodeficiency virus type 1 gp120. AB - Cell-mediated immunity depends in part on appropriate migration and localization of cytotoxic T lymphocytes (CTL), a process regulated by chemokines and adhesion molecules. Many viruses, including human immunodeficiency virus type 1 (HIV-1), encode chemotactically active proteins, suggesting that dysregulation of immune cell trafficking may be a strategy for immune evasion. HIV-1 gp120, a retroviral envelope protein, has been shown to act as a T-cell chemoattractant via binding to the chemokine receptor and HIV-1 coreceptor CXCR4. We have previously shown that T cells move away from the chemokine stromal cell-derived factor 1 (SDF-1) in a concentration-dependent and CXCR4 receptor-mediated manner. Here, we demonstrate that CXCR4-binding HIV-1 X4 gp120 causes the movement of T cells, including HIV-specific CTL, away from high concentrations of the viral protein. This migratory response is CD4 independent and inhibited by anti-CXCR4 antibodies and pertussis toxin. Additionally, the expression of X4 gp120 by target cells reduces CTL efficacy in an in vitro system designed to account for the effect of cell migration on the ability of CTL to kill their target cells. Recombinant X4 gp120 also significantly reduced antigen-specific T-cell infiltration at a site of antigen challenge in vivo. The repellant activity of HIV-1 gp120 on immune cells in vitro and in vivo was shown to be dependent on the V2 and V3 loops of HIV-1 gp120. These data suggest that the active movement of T cells away from CXCR4-binding HIV-1 gp120, which we previously termed fugetaxis, may provide a novel mechanism by which HIV-1 evades challenge by immune effector cells in vivo. PMID- 15113901 TI - Acute infection with Epstein-Barr virus targets and overwhelms the peripheral memory B-cell compartment with resting, latently infected cells. AB - In this paper we demonstrate that during acute infection with Epstein-Barr virus (EBV), the peripheral blood fills up with latently infected, resting memory B cells to the point where up to 50% of all the memory cells may carry EBV. Despite this massive invasion of the memory compartment, the virus remains tightly restricted to memory cells, such that, in one donor, fewer than 1 in 10(4) infected cells were found in the naive compartment. We conclude that, even during acute infection, EBV persistence is tightly regulated. This result confirms the prediction that during the early phase of infection, before cellular immunity is effective, there is nothing to prevent amplification of the viral cycle of infection, differentiation, and reactivation, causing the peripheral memory compartment to fill up with latently infected cells. Subsequently, there is a rapid decline in infected cells for the first few weeks that approximates the decay in the cytotoxic-T-cell responses to viral replicative antigens. This phase is followed by a slower decline that, even by 1 year, had not reached a steady state. Therefore, EBV may approach but never reach a stable equilibrium. PMID- 15113902 TI - The V1/V2 domain of gp120 is a global regulator of the sensitivity of primary human immunodeficiency virus type 1 isolates to neutralization by antibodies commonly induced upon infection. AB - A major problem hampering the development of an effective vaccine against human immunodeficiency virus type 1 (HIV-1) is the resistance of many primary viral isolates to antibody-mediated neutralization. To identify factors responsible for this resistance, determinants of the large differences in neutralization sensitivities of HIV-1 pseudotyped with Env proteins derived from two prototypic clade B primary isolates were mapped. SF162 Env pseudotypes were neutralized very potently by a panel of sera from HIV-infected individuals, while JR-FL Env pseudotypes were neutralized by only a small fraction of these sera. This differential sensitivity to neutralization was also observed for a number of monoclonal antibodies (MAbs) directed against sites in the V2, V3, and CD4 binding domains, despite often similar binding affinities of these MAbs towards the two soluble rgp120s. The neutralization phenotypes were switched for chimeric Envs in which the V1/V2 domains of these two sequences were exchanged, indicating that the V1/V2 region regulated the overall neutralization sensitivity of these Envs. These results suggested that the inherent neutralization resistance of JR FL, and presumably of related primary isolates, is to a great extent mediated by gp120 V1/V2 domain structure rather than by sequence variations at the target sites. Three MAbs (immunoglobulin G-b12, 2G12, and 2F5) previously reported to possess broad neutralizing activity for primary HIV-1 isolates neutralized JR-FL virus at least as well as SF162 virus and were not significantly affected by the V1/V2 domain exchanges. The rare antibodies capable of neutralizing a broad range of primary isolates thus appeared to be targeted to exceptional epitopes that are not sensitive to V1/V2 domain regulation of neutralization sensitivity. PMID- 15113903 TI - A mutation in the HLA-B*2705-restricted NP383-391 epitope affects the human influenza A virus-specific cytotoxic T-lymphocyte response in vitro. AB - Viruses can exploit a variety of strategies to evade immune surveillance by cytotoxic T lymphocytes (CTL), including the acquisition of mutations in or adjacent to CTL epitopes. Recently, an amino acid substitution (R384G) in an HLA B*2705-restricted CTL epitope in the influenza A virus nucleoprotein (nucleoprotein containing residues 383 to 391 [NP(383-391)]; SRYWAIRTR, where R is the residue that was mutated) was associated with escape from CTL-mediated immunity. The effect of this mutation on the in vitro influenza A virus-specific CTL response was studied. To this end, two influenza A viruses, one with and one without the NP(383-391) epitope, were constructed by reverse genetics and designated influenza viruses A/NL/94-384R and A/NL/94-384G, respectively. The absence of the HLA-B*2705-restricted CTL epitope in influenza virus A/NL/94-384G was confirmed by using (51)Cr release assays with a T-cell clone specific for the NP(383-391) epitope. In addition, peripheral blood mononuclear cells (PBMC) stimulated with influenza virus A/NL/94-384G failed to recognize HLA-B*2705 positive target cells pulsed with the original NP(383-391) peptide. The proportion of virus-specific CD8+ gamma interferon (IFN-gamma)-positive T cells in in vitro-stimulated PBMC was determined by intracellular IFN-gamma staining after restimulation with virus-infected autologous B-lymphoblastoid cell lines and C1R cell lines expressing only HLA-B*2705. The proportion of virus-specific CD8+ T cells was lower in PBMC stimulated in vitro with influenza virus A/NL/94 384G obtained from several HLA-B*2705-positive donors than in PBMC stimulated with influenza virus A/NL/94-384R. This finding indicated that amino acid variations in CTL epitopes can affect the virus-specific CTL response and that the NP(383-391) epitope is the most important HLA-B*2705-restricted epitope in the nucleoprotein of influenza A viruses. PMID- 15113904 TI - Human immunodeficiency virus type 1 activates plasmacytoid dendritic cells and concomitantly induces the bystander maturation of myeloid dendritic cells. AB - In this study, we analyzed the phenotypic and physiological consequences of the interaction of plasmacytoid dendritic cells (pDCs) with human immunodeficiency virus type 1 (HIV-1). pDCs are one cellular target of HIV-1 and respond to the virus by producing alpha/beta interferon (IFN-alpha/beta) and chemokines. The outcome of this interaction, notably on the function of bystander myeloid DC (CD11c+ DCs), remains unclear. We therefore evaluated the effects of HIV-1 exposure on these two DC subsets under various conditions. Blood-purified pDCs and CD11c+ DCs were exposed in vitro to HIV-1, after which maturation markers, cytokine production, migratory capacity, and CD4 T-cell stimulatory capacity were analyzed. pDCs exposed to different strains of infectious or even chemically inactivated, nonreplicating HIV-1 strongly upregulated the expression of maturation markers, such as CD83 and functional CCR7, analogous to exposure to R 848, a synthetic agonist of toll-like receptor-7 and -8. In addition, HIV-1 activated pDCs produced cytokines (IFN-alpha and tumor necrosis factor alpha), migrated in response to CCL19 and, in coculture, matured CD11c+ DCs, which are not directly activated by HIV. pDCs also acquired the ability to stimulate naive CD4+ T cells, albeit less efficiently than CD11c+ DCs. This HIV-1-induced maturation of both DC subsets may explain their disappearance from the blood of patients with high viral loads and may have important consequences on HIV-1 cellular transmission and HIV-1-specific T-cell responses. PMID- 15113905 TI - Analysis of host range phenotypes of primate hepadnaviruses by in vitro infections of hepatitis D virus pseudotypes. AB - Hepatitis B virus (HBV) and woolly monkey hepatitis B virus (WMHBV) have natural host ranges that are limited to closely related species. The barrier for infection of primates seems to be at the adsorption and/or entry steps of the viral replication cycle, since a human hepatoma cell line is permissive for HBV and WMHBV replication following transfection of cloned DNA. We hypothesized that the HBV and WMHBV envelope proteins contain the principal viral determinants of host range. As previously shown by using the hepatitis D virus (HDV) system, recombinant HBV-HDV particles were infectious in chimpanzee as well as human hepatocytes. We extended the HDV system to include HDV particles pseudotyped with the WMHBV envelope. In agreement with the natural host ranges of HBV and WMHBV, in vitro infections demonstrated that HBV-HDV and WM-HDV particles preferentially infected human and spider monkey cells, respectively. Previous studies have implicated the pre-S1 region of the large (L) envelope protein in receptor binding and host range; therefore, recombinant HDV particles were pseudotyped with the hepadnaviral envelopes containing chimeric L proteins with the first 40 amino acids from the pre-S1 domain exchanged between HBV and WMHBV. Surprisingly, addition of the human amino terminus to the WMHBV L protein increased infectivity on spider monkey hepatocytes but did not increase infectivity for human hepatocytes. Based upon these data, we discuss the possibility that the L protein may be comprised of two domains that affect infectivity and that sequences downstream of residue 40 may influence host range and receptor binding or entry. PMID- 15113906 TI - CD4+ T cells from CD4C/HIVNef transgenic mice show enhanced activation in vivo with impaired proliferation in vitro but are dispensable for the development of a severe AIDS-like organ disease. AB - The cellular and molecular mechanisms of dysfunction and depletion of CD4+ T lymphocytes over the course of human immunodeficiency virus type 1 (HIV-1) infection are still incompletely understood, but chronic immune activation is thought to play an important role in disease progression. We studied CD4+ T-cell biology in CD4C/HIV transgenic (Tg) mice, in which Nef expression is sufficient to induce a severe AIDS-like disease including a preferential decrease of CD4+ T cells. We show here that Nef-expressing Tg CD4+ T cells exhibit an activated/memory-like phenotype which appears to be independent of antigenic stimulation, as documented in experiments involving breeding with AD10 TcR Tg mice. In addition, in vivo bromodeoxyuridine incorporation showed that a larger proportion of Tg than non-Tg CD4+ T cells entered the S phase. However, in vitro, Tg CD4+ T cells were found to have a very limited capacity to divide in response to stimulation with anti-CD3 and anti-CD28 or in allogeneic mixed leukocyte reactions. Interestingly, despite these observations, the deletion of Tg CD4+ T cells had little impact on the development of other AIDS-like organ phenotypes. Thus, the Nef-induced chronic activation of CD4+ T cells may exhaust the T-cell pool and may contribute to the thymic atrophy and the low number of CD4+ T cells observed in these Tg mice. PMID- 15113907 TI - Role of transmembrane domain and cytoplasmic tail amino acid sequences of influenza a virus neuraminidase in raft association and virus budding. AB - Influenza virus neuraminidase (NA), a type II transmembrane glycoprotein, possesses receptor-destroying activity and thereby facilitates virus release from the cell surface. Among the influenza A viruses, both the cytoplasmic tail (CT) and transmembrane domain (TMD) amino acid sequences of NA are highly conserved, yet their function(s) in virus biology remains unknown. To investigate the role of amino acid sequences of the CT and TMD on the virus life cycle, we systematically mutagenized the entire CT and TMD of NA by converting two to five contiguous amino acids to alanine. In addition, we also made two chimeric NA by replacing the CT proximal one-third amino acids of the NA TMD [NA(1T2N)NA] and the entire NA TMD (NATRNA) with that of human transferrin receptor (TR) (a type II transmembrane glycoprotein). We rescued transfectant mutant viruses by reverse genetics and examined their phenotypes. Our results show that all mutated and chimeric NAs could be rescued into transfectant viruses. Different mutants showed pleiotropic effects on virus growth and replication. Some mutants (NA2A5, NA3A7, and NA4A10) had little effect on virus growth while others (NA3A2, NA5A27, and NA5A31) produced about 50- to 100-fold-less infectious virus and still some others (NA5A14, NA4A19, and NA4A23) exhibited an intermediate phenotype. In general, mutations towards the ectodomain-proximal sequences of TMD progressively caused reduction in NA enzyme activity, affected lipid raft association, and attenuated virus growth. Electron microscopic analysis showed that these mutant viruses remained aggregated and bound to infected cell surfaces and could be released from the infected cells by bacterial NA treatment. Moreover, viruses containing mutations in the extreme N terminus of the CT (NA3A2) as well as chimeric NA containing the TMD replaced partially [NA(1T2N)NA] or fully (NATRNA) with TR TMD caused reduction in virus growth and exhibited the morphological phenotype of elongated particles. These results show that although the sequences of NA CT and TMD per se are not absolutely essential for the virus life cycle, specific amino acid sequences play a critical role in providing structural stability, enzyme activity, and lipid raft association of NA. In addition, aberrant morphogenesis including elongated particle formation of some mutant viruses indicates the involvement of NA in virus morphogenesis and budding. PMID- 15113908 TI - Immunogenicity of constrained monoclonal antibody A32-human immunodeficiency virus (HIV) Env gp120 complexes compared to that of recombinant HIV type 1 gp120 envelope glycoproteins. AB - One strategy for the generation of broadly reactive neutralizing antibodies (NA) against human immunodeficiency virus type 1 (HIV-1) primary isolates is to use immunogens that have constrained HIV-1 envelope gp120 conformations reflective of triggered envelope on the surface of virions. A major change in gp120 following binding to CD4 is the enhanced exposure of the CCR5 binding site. One inducer of CCR5 binding site epitopes on gp120 is the human anti-gp120 monoclonal antibody, A32. We have made cross-linked A32-rgp120(89.6) and A32-rgp120(BaL) complexes and have compared their immunogenicities to those of uncomplexed recombinant gp120(BaL) (rgp120(BaL)) and rgp120(89.6). A32-rgp120(89.6) and A32-rgp120(BaL) complexes had stable induced CCR5 binding site expression compared to that of uncomplexed rgp120s. However, the A32-rgp120 complexes had similar capacities in guinea pigs for induction of NA against HIV-1 primary isolates versus that of rgp120 alone. A32-rgp120(89.6) induced antibodies that neutralized 6 out of 11 HIV-1 isolates, while rgp120(89.6) alone induced antibodies that neutralized 4 out of 11 HIV-1 isolates. A32-rgp120(BaL) complexes induced antibodies that neutralized 4 out of 14 HIV-1 isolates while, surprisingly, non-cross-linked rgp120(BaL) induced antibodies that neutralized 9 out of 14 (64%) HIV-1 isolates. Thus, stable enhanced expression of the coreceptor binding site on constrained gp120 is not sufficient for inducing broadly neutralizing anti-HIV-1 NA. Moreover, the ability of HIV-1 rgp120(BaL) to induce antibodies that neutralized approximately 60% of subtype B HIV-1 isolates warrants consideration of using HIV 1 BaL as a starting point for immunogen design for subtype B HIV-1 experimental immunogens. PMID- 15113909 TI - Copatching and lipid raft association of different viral glycoproteins expressed on the surfaces of pseudorabies virus-infected cells. AB - Pseudorabies virus (PRV) is a swine alphaherpesvirus that is closely related to human herpes simplex virus (HSV). Both PRV and HSV express a variety of viral envelope glycoproteins in the plasma membranes of infected cells. Here we show that at least four major PRV glycoproteins (gB, gC, gD, and gE) in the plasma membrane of infected swine kidney cells and monocytes seem to be linked, since monospecific antibody-induced patching of any one of these proteins results in copatching of the others. Further, for all four PRV glycoproteins, monospecific antibody-induced patches were enriched in GM1, a typical marker of lipid raft microdomains, but were excluded for transferrin receptor, a nonraft marker, suggesting that these viral proteins may associate with lipid rafts. However, only gB and, to a lesser extent, gE were found in lipid raft fractions by using detergent floatation assays, indicating that gC and gD do not show strong lipid raft association. Addition of methyl-beta-cyclodextrin (MCD), a cholesterol depleting agent that is commonly used to disrupt lipid rafts, only slightly reduced copatching efficiency between the different viral proteins, indicating that other factors, perhaps tegument-glycoprotein interactions, may be important for the observed copatching events. On the other hand, MCD strongly reduced polarization of the antibody-induced viral glycoprotein patches to a cap structure, a gE-dependent process that has been described for specific PRV- and HSV-infected cells. Therefore, we hypothesize that efficient gE-mediated capping of antibody-antigen patches may require the lipid raft-associated signal transduction machinery. PMID- 15113910 TI - Cellular RNA helicase p68 relocalization and interaction with the hepatitis C virus (HCV) NS5B protein and the potential role of p68 in HCV RNA replication. AB - Chronic infection by hepatitis C virus (HCV) can lead to severe hepatitis and cirrhosis and is closely associated with hepatocellular carcinoma. The replication cycle of HCV is poorly understood but is likely to involve interaction with host factors. In this report, we show that NS5B, the HCV RNA dependent RNA polymerase (RdRp), interacts with a human RNA helicase, p68. Transient expression of NS5B alone, as well as the stable expression of all the nonstructural proteins in a HCV replicon-bearing cell line (V. Lohmann, F. Korner, J.-O. Koch, U. Herian, L. Theilmann, and R. Bartenschlager, Science 285:110-113), causes the redistribution of endogenous p68 from the nucleus to the cytoplasm. Deletion of the C-terminal two-thirds of NS5B (NS5BDeltaC) dramatically reduces its coimmunoprecipitation (co-IP) with endogenous p68, while the deletion of the N-terminal region (NS5BDeltaN1 and NS5BDeltaN2) does not affect its interaction with p68. In consistency with the co-IP results, NS5BDeltaC does not cause the relocalization of p68 whereas NS5BDeltaN1 does. With a replicon cell line, we were not able to detect a change in positive- and negative-strand synthesis when p68 levels were reduced using small interfering RNA (siRNA). In cells transiently transfected with a full-length HCV construct, however, the depletion (using specific p68 siRNA) of endogenous p68 correlated with a reduction in the transcription of negative-strand from positive-strand HCV RNA. Overexpression of NS5B and NS5BDeltaN1, but not that of NS5BDeltaC, causes a reduction in the negative-strand synthesis, indicating that overexpressed NS5B and NS5BDeltaN1 sequesters p68 from the replication complexes (thus reducing their replication activity levels). Identification of p68 as a cellular factor involved in HCV replication, at least for cells transiently transfected with a HCV expression construct, is a step towards understanding HCV replication. PMID- 15113911 TI - Mutated form of the Newcastle disease virus hemagglutinin-neuraminidase interacts with the homologous fusion protein despite deficiencies in both receptor recognition and fusion promotion. AB - The Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) protein mediates attachment to cellular receptors. The fusion (F) protein promotes viral entry and spread. However, fusion is dependent on a virus-specific interaction between the two proteins that can be detected at the cell surface by a coimmunoprecipitation assay. A point mutation of I175E in the neuraminidase (NA) active site converts the HN of the Australia-Victoria isolate of the virus to a form that can interact with the F protein despite negligible receptor recognition and fusion-promoting activities. Thus, I175E-HN could represent a fusion intermediate in which HN and F are associated and primed for the promotion of fusion. Both the attachment and fusion-promoting activities of this mutant HN protein can be rescued either by NA activity contributed by another HN protein or by a set of four substitutions at the dimer interface. These substitutions were identified by the evaluation of chimeras composed of segments from HN proteins derived from two different NDV strains. These findings suggest that the I175E substitution converts HN to an F-interactive form, but it is one for which receptor binding is still required for fusion promotion. The data also indicate that the integrity of the HN dimer interface is critical to its receptor recognition activity. PMID- 15113912 TI - CD8+-cell-mediated suppression of virulent simian immunodeficiency virus during tenofovir treatment. AB - The ability of tenofovir to suppress viremia in simian immunodeficiency virus (SIV)-infected macaques for years despite the presence of virulent viral mutants with reduced in vitro susceptibility is unprecedented in this animal model. In vivo cell depletion experiments demonstrate that tenofovir's ability to suppress viremia during acute and chronic infection is significantly dependent on the presence of CD8+ lymphocytes. Continuous tenofovir treatment was required to maintain low viremia. Although it is unclear whether this immune-mediated suppression of viremia is linked to tenofovir's direct antiviral efficacy or is due to independent immunomodulatory effects, these studies prove the concept that antiviral immune responses can play a crucial role in suppressing viremia during anti-human immunodeficiency virus drug therapy. PMID- 15113913 TI - The papillomavirus E7 oncoprotein is ubiquitinated by UbcH7 and Cullin 1- and Skp2-containing E3 ligase. AB - Recurrent infections with high-risk human papillomaviruses (HPVs) are associated with human cervical cancers. All HPV-associated cancer tissues express the viral oncoproteins E6 and E7, which stimulate cell growth. The expression of E7 is crucial for both the initiation and the maintenance of HPV-associated cancer. Recent studies showed that the level of E7 in cancer cells is regulated by ubiquitin-dependent proteolysis through the 26S proteasome. In this study, we characterized the enzymes involved in the ubiquitin-dependent proteolysis of E7. We show that UbcH7, an E2 ubiquitin-conjugating enzyme, is specifically involved in the ubiquitination of E7. Furthermore, we show that E7 interacts with the SCF (Skp-Cullin-F box) ubiquitin ligase complex containing Cullin 1 (Cul1) and Skp2 and can be ubiquitinated by the Cul1-containing ubiquitin ligase in vitro. Coimmunoprecipitation analyses revealed that E7 interacts with Skp2 and Cul1 in vivo. Finally, the half-life of E7 was found to be significantly longer in Skp2( /-) mouse embryo fibroblasts (MEFs) than in wild-type MEFs. Taken together, these results suggest that the Cul1- and Skp2-containing ubiquitin ligase plays a role in the ubiquitination and proteolysis of E7. In HPV type 16-containing cervical carcinoma cell line Caski, E7 localizes to both the cytoplasm and the nucleus. Brief treatment of Caski cells with MG132 (a proteasome inhibitor) causes the accumulation of E7 in discrete nuclear bodies. These nuclear bodies are detergent insoluble and contain polyubiquitinated E7. We suggest that E7 relocates to specific nuclear bodies for proteolysis in HPV-containing epithelial cells. PMID- 15113914 TI - The herpes simplex virus type 1 UL20 protein modulates membrane fusion events during cytoplasmic virion morphogenesis and virus-induced cell fusion. AB - The herpes simplex virus type 1 (HSV-1) UL20 protein is an important determinant for virion morphogenesis and virus-induced cell fusion. A precise deletion of the UL20 gene in the HSV-1 KOS strain was constructed without affecting the adjacent UL20.5 gene. The resultant KOS/UL20-null virus produced small plaques of 8 to 15 cells in Vero cells while it produced wild-type plaques on the complementing cell line G5. Electron microscopic examination of infected cells revealed that the KOS/UL20-null virions predominantly accumulated capsids in the cytoplasm while a small percentage of virions were found as enveloped virions within cytoplasmic vacuoles. Recently, it was shown that UL20 expression was necessary and sufficient for cell surface expression of gK (T. P. Foster, X. Alvarez, and K. G. Kousoulas, J. Virol. 77:499-510, 2003). Therefore, we investigated the effect of UL20 on virus-induced cell fusion caused by syncytial mutations in gB and gK by constructing recombinant viruses containing the gBsyn3 or gKsyn1 mutations in a UL20-null genetic background. Both recombinant viruses failed to cause virus induced cell fusion in Vero cells while they readily caused fusion of UL20-null complementing G5 cells. Ultrastructural examination of UL20-null viruses carrying the gBsyn3 or gKsyn1 mutation revealed a similar distribution of virions as the KOS/UL20-null virus. However, cytoplasmic vacuoles contained aberrant virions having multiple capsids within a single envelope. These multicapsid virions may have been formed either by fusion of viral envelopes or by the concurrent reenvelopment of multiple capsids. These results suggest that the UL20 protein regulates membrane fusion phenomena involved in virion morphogenesis and virus induced cell fusion. PMID- 15113915 TI - Identification of the nuclear export signal and STAT-binding domains of the Nipah virus V protein reveals mechanisms underlying interferon evasion. AB - The V proteins of Nipah virus and Hendra virus have been demonstrated to bind to cellular STAT1 and STAT2 proteins to form high-molecular-weight complexes that inhibit interferon (IFN)-induced antiviral transcription by preventing STAT nuclear accumulation. Analysis of the Nipah virus V protein has revealed a region between amino acids 174 and 192 that functions as a CRM1-dependent nuclear export signal (NES). This peptide is sufficient to complement an export-defective human immunodeficiency virus Rev protein, and deletion and substitution mutagenesis revealed that this peptide is necessary for both V protein shuttling and cytoplasmic retention of STAT1 and STAT2 proteins. However, the NES is not required for V-dependent IFN signaling inhibition. IFN signaling is blocked primarily by interaction between Nipah virus V residues 100 to 160 and STAT1 residues 509 to 712. Interaction with STAT2 requires a larger Nipah virus V segment between amino acids 100 and 300, but deletion of residues 230 to 237 greatly reduced STAT2 coprecipitation. Further, V protein interactions with cellular STAT1 is a prerequisite for STAT2 binding, and sequential immunoprecipitations demonstrate that V, STAT1, and STAT2 can form a tripartite complex. These findings characterize essential regions for Henipavirus V proteins that represent potential targets for therapeutic intervention. PMID- 15113916 TI - Analysis of adenovirus sequestration in the liver, transduction of hepatic cells, and innate toxicity after injection of fiber-modified vectors. AB - After intravenous administration, adenovirus (Ad) vectors are predominantly sequestered by the liver. Delineating the mechanisms for Ad accumulation in the liver is crucial for a better understanding of Ad clearance and Ad-associated innate toxicity. To help address these issues, in this study, we used Ad vectors with different fiber shaft lengths and either coxsackievirus-Ad receptor (CAR) interacting Ad serotype 9 (Ad9) or non-CAR-interacting Ad35 fiber knob domains. We analyzed the kinetics of Ad vector accumulation in the liver, uptake into hepatocytes and Kupffer cells, and induction of cytokine expression and release in response to systemic vector application. Immediately after intravenous injection, all Ad vectors accumulated equally efficiently in the liver; however, only genomes of long-shafted Ads were maintained in the liver tissue over time. We found that Kupffer cell uptake of long-shafted Ads was mediated by the fiber knob domain and was CAR independent. The short-shafted Ads were unable to efficiently interact with hepatocellular receptors and were not taken up by Kupffer cells. Moreover, our studies indicated that Kupffer cells were not the major reservoir for the observed accumulation of Ads (used in this study) in the liver within the first 30 min after virus infusion. The lower level of liver cell transduction by short-shafted Ads correlated with a significantly reduced inflammatory anti-Ad response as well as liver damage induced by the systemic administration of these vectors. This study contributes to a better understanding of the biology of systemically applied Ad and will help in designing safer vectors that can efficiently transduce target tissues. PMID- 15113917 TI - Rice dwarf phytoreovirus segment S6-encoded nonstructural protein has a cell-to cell movement function. AB - Rice dwarf virus (RDV) is a member of the genus Phytoreovirus, which is composed of viruses with segmented double-stranded RNA genomes. Proteins that support the intercellular movement of these viruses in the host have not been identified. Microprojectile bombardment was used to determine which open reading frames (ORFs) support intercellular movement of a heterologous virus. A plasmid containing an infectious clone of Potato virus X (PVX) defective in cell-to-cell movement and expressing either beta-glucuronidase or green fluorescent protein (GFP) was used for cobombardment with plasmids containing ORFs from RDV gene segments S1 through S12 onto leaves of Nicotiana benthamiana. Cell-to-cell movement of the movement-defective PVX was restored by cobombardment with a plasmid containing S6. In the absence of S6, no other gene segment supported movement. Identical results were obtained with Nicotiana tabacum, a host that allows fewer viruses to infect and spread within its tissue. S6 supported the cell-to-cell movement of the movement-defective PVX in sink and source leaves of N. benthamiana. A mutant S6 lacking the translation start codon did not complement the cell-to-cell movement of the movement-defective PVX. An S6 protein product (Pns6)-enhanced GFP fusion was observed near or within cell walls of epidermal cells from N. tabacum. By immunocytochemistry, unfused Pns6 was localized to plasmodesmata in rice leaves infected with RDV. S6 thus encodes a protein with characteristics identical to those of other viral proteins required for the cell-to-cell movement of their genome and therefore is likely required for the cell-to-cell movement of RDV. PMID- 15113918 TI - Characterization of a subtype D human immunodeficiency virus type 1 isolate that was obtained from an untreated individual and that is highly resistant to nonnucleoside reverse transcriptase inhibitors. AB - Human immunodeficiency virus type 1 (HIV-1) isolates derived from HIV-infected, treatment-naive Ugandan infants were propagated and tested for sensitivity to antiretroviral (ARV) drugs. Although most subtype A and D isolates displayed inhibition profiles similar to those of subtype B strains, a subtype D isolate identified as D14-UG displayed high-level resistance to nevirapine in peripheral blood mononuclear cell cultures (>2,000-fold) and in MT4 cell cultures ( approximately 800-fold) but weaker resistance to delavirdine ( approximately 13 fold) and efavirenz ( approximately 8-fold) in MT4 cell cultures. To investigate the possible mechanism for this resistance to nonnucleoside reverse transcriptase (RT) inhibitors (NNRTIs), the RT coding region in pol was sequenced and compared to the consensus RT sequence of NNRTI-resistant and NNRTI-sensitive subtype A, B, and D HIV-1 isolates. D14-UG did not contain the classic amino acid substitutions conferring NNRTI resistance (e.g., Y181C, K103N, and G190A) but did have some putative sites associated with drug resistance, I135L, T139V, and V245T. Wild type and mutated protease-RT genes from D14-UG and an NNRTI-sensitive subtype D isolate from Uganda (D13-UG) were cloned into pNL4-3 to produce recombinant viruses and to determine the effects of the mutations on susceptibility to ARV drugs, specifically, NNRTIs. The results showed that I135L and/or V245T mutations can confer high-level resistance to nevirapine and delavirdine as well as low level cross-resistance to efavirenz. Finally, ex vivo fitness analyses suggested that NNRTI-resistant sites 135L and 245T in wild-type isolate D14-UG may reduce RT fitness but do not have an impact on the fitness of the primary HIV-1 isolate. PMID- 15113919 TI - Frequent dual initiation in human immunodeficiency virus-based vectors containing two primer-binding sites: a quantitative in vivo assay for function of initiation complexes. AB - We previously demonstrated that murine leukemia virus (MLV)-based vectors containing two primer-binding sites (PBSs) have the capacity to initiate reverse transcription more than once (Y. A. Voronin and V. K. Pathak, Virology 312:281 294, 2003). To determine whether human immunodeficiency virus (HIV)-based vectors also have the capacity to initiate reverse transcription twice, we constructed an HIV type 1 (HIV-1)-based vector containing the HIV-1 PBS, a green fluorescent protein reporter gene (GFP), and a second PBS derived from HIV-2 3' of GFP. Simultaneous initiation of reverse transcription at both the 5' HIV-1 PBS and 3' HIV-2 PBS was predicted to result in deletion of GFP. As in the MLV-based vectors, GFP was deleted in approximately 25% of all proviruses, indicating frequent dual initiation in HIV-based vectors containing two PBSs. Quantitative real-time PCR analysis of early reverse transcription products indicated that HIV 1 reverse transcriptase efficiently used the HIV-2 PBS. To investigate tRNA primer-RNA template interactions in vivo, we introduced several mutations in the HIV-2 U5 region. The effects of these mutations on the efficiency of reverse transcription initiation were measured by quantitative real-time PCR analysis of early reverse transcription products, with initiation at the HIV-1 PBS used as an internal control. Disruption of the lower and upper parts of the U5-inverted repeat stem reduced the efficiency of initiation 20- and 6-fold, respectively. In addition, disruption of the proposed interactions between viral RNA and tRNA(Lys3) thymidine-pseudouridine-cytidine and anticodon loops decreased the efficiency of initiation seven- and sixfold, respectively. These results demonstrate the relative influence of various RNA-RNA interactions on the efficiency of initiation in vivo. Furthermore, the two-PBS vector system provides a sensitive and quantitative in vivo assay for analysis of RNA-RNA and protein RNA interactions that can influence the efficiency of reverse transcription initiation. PMID- 15113920 TI - Analysis of N-linked glycosylation of hantaan virus glycoproteins and the role of oligosaccharide side chains in protein folding and intracellular trafficking. AB - The membrane glycoproteins Gn and Gc of Hantaan virus (HTNV) (family Bunyaviridae) are modified by N-linked glycosylation. The glycoproteins contain six potential sites for the attachment of N-linked oligosaccharides, five sites on Gn and one on Gc. The properties of the N-linked oligosaccharide chains were analyzed by treatment with endoglycosidase H, peptide:N-glycosidase F, tunicamycin, and deoxynojirimycin and were confirmed to be completely of the high mannose type. Ten glycoprotein gene mutants were constructed by site-directed mutagenesis, including six single N glycosylation site mutants and four double site mutants. We determined that four sites (N134, -235, -347, and -399) on Gn and the only site (N928) on Gc in their ectodomains are utilized, whereas the fifth site on Gn (N609), which faces the cytoplasm, is not glycosylated. The importance of individual N-oligosaccharide chains varied with respect to folding and intracellular transport. The oligosaccharide chain on residue N134 was found to be crucial for protein folding, whereas single mutations at the other glycosylation sites were better tolerated. Mutation at glycosylation sites N235 and N399 together resulted in Gn misfolding. The endoplasmic reticulum chaperones calnexin and calreticulin were found to be involved in HTNV glycoprotein folding. Our data demonstrate that N-linked glycosylation of HTNV glycoproteins plays important and differential roles in protein folding and intracellular trafficking. PMID- 15113921 TI - Binding and susceptibility to postentry restriction factors in monkey cells are specified by distinct regions of the human immunodeficiency virus type 1 capsid. AB - In cells of Old World and some New World monkeys, dominant factors restrict human immunodeficiency virus type 1 (HIV-1) infections after virus entry. The simian immunodeficiency virus SIV(mac) is less susceptible to these restrictions, a property that is determined largely by the viral capsid protein. For this study, we altered exposed amino acid residues on the surface of the HIV-1 capsid, changing them to the corresponding residues found on the SIV(mac) capsid. We identified two distinct pathways of escape from early, postentry restriction in monkey cells. One set of mutants that were altered near the base of the cyclophilin A-binding loop of the N-terminal capsid domain or in the interdomain linker exhibited a decreased ability to bind the restricting factor(s). Consistent with the location of this putative factor-binding site, cyclophilin A and the restricting factor(s) cooperated to achieve the postentry block. A second set of mutants that were altered in the ridge formed by helices 3 and 6 of the N terminal capsid domain efficiently bound the restricting factor(s) but were resistant to the consequences of factor binding. These results imply that binding of the simian restricting factor(s) is not sufficient to mediate the postentry block to HIV-1 and that SIV(mac) capsids escape the block by decreases in both factor binding and susceptibility to the effects of the factor(s). PMID- 15113922 TI - Identification of a novel bovine herpesvirus 1 transcript containing a small open reading frame that is expressed in trigeminal ganglia of latently infected cattle. AB - Bovine herpesvirus 1 (BHV-1), like other Alphaherpesvirinae subfamily members, establishes latency in sensory neurons. The latency-related (LR) RNA is abundantly expressed during latency, and expression of an LR protein is required for the latency reactivation cycle in cattle. Within LR promoter sequences, a 135 amino-acid open reading frame (ORF) was identified, ORF-E, that is antisense to the LR RNA. ORF-E is also downstream of the gene encoding the major viral transcriptional activator, bICP0. Strand-specific reverse transcription-PCR demonstrated that a transcript containing ORF-E was consistently expressed in trigeminal ganglia (TG) of latently infected calves, productively infected cultured cells, and acutely infected calves. As expected, a late transcript encoding glycoprotein C was not detected in TG of latently infected calves. The ORF-E transcript is polyadenylated and is expressed early when cultured bovine cells are productively infected. Protein coding sequences containing ORF-E were fused to green fluorescent protein (GFP) to examine the cellular localization of the putative protein. In transiently transfected mouse neuroblastoma (neuro-2A) and human neuroblastoma (SK-N-SH) cells, the ORF-E/GFP fusion protein was detected in discreet domains within the nucleus. In contrast, the ORF-E/GFP fusion protein was detected in the cytoplasm and nucleus of rabbit skin cells and bovine kidney cells. As expected, the GFP protein was expressed in the cytoplasm and nucleus of transfected cells. These studies indicate that the ORF-E transcript is consistently expressed during latency. We suggest that the ORF-E gene regulates some aspect of the latency reactivation cycle. PMID- 15113923 TI - CD4-induced T-20 binding to human immunodeficiency virus type 1 gp120 blocks interaction with the CXCR4 coreceptor. AB - The synthetic peptide T-20, which corresponds to a sequence within the C-terminal heptad repeat region (HR2) of the human immunodeficiency virus type 1 (HIV-1) gp41 envelope glycoprotein, potently inhibits viral membrane fusion and entry. Although T-20 is thought to bind the N-terminal heptad repeat region (HR1) of gp41 and interfere with gp41 conformational changes required for membrane fusion, coreceptor specificity determined by the V3 loop of gp120 strongly influences the sensitivity of HIV-1 variants to T-20. Here, we show that T-20 binds to the gp120 glycoproteins of HIV-1 isolates that utilize CXCR4 as a coreceptor in a manner determined by the sequences of the gp120 V3 loop. T-20 binding to gp120 was enhanced in the presence of soluble CD4. Analysis of T-20 binding to gp120 mutants with variable loop deletions and the reciprocal competition of T-20 and particular anti-gp120 antibodies suggested that T-20 interacts with a gp120 region near the base of the V3 loop. Consistent with the involvement of this region in coreceptor binding, T-20 was able to block the interaction of gp120-CD4 complexes with the CXCR4 coreceptor. These results help to explain the increased sensitivity of CXCR4-specific HIV-1 isolates to the T-20 peptide. Interactions between the gp41 HR2 region and coreceptor-binding regions of gp120 may also play a role in the function of the HIV-1 envelope glycoproteins. PMID- 15113924 TI - Properties of replication-competent vesicular stomatitis virus vectors expressing glycoproteins of filoviruses and arenaviruses. AB - Replication-competent recombinant vesicular stomatitis viruses (rVSVs) expressing the type I transmembrane glycoproteins and selected soluble glycoproteins of several viral hemorrhagic fever agents (Marburg virus, Ebola virus, and Lassa virus) were generated and characterized. All recombinant viruses exhibited rhabdovirus morphology and replicated cytolytically in tissue culture. Unlike the rVSVs with an additional transcription unit expressing the soluble glycoproteins, the viruses carrying the foreign transmembrane glycoproteins in replacement of the VSV glycoprotein were slightly attenuated in growth. Biosynthesis and processing of the foreign glycoproteins were authentic, and the cell tropism was defined by the transmembrane glycoprotein. None of the rVSVs displayed pathogenic potential in animals. The rVSV expressing the Zaire Ebola virus transmembrane glycoprotein mediated protection in mice against a lethal Zaire Ebola virus challenge. Our data suggest that the recombinant VSV can be used to study the role of the viral glycoproteins in virus replication, immune response, and pathogenesis. PMID- 15113925 TI - Regional differences in viral growth and central nervous system injury correlate with apoptosis. AB - Infection of neonatal mice with reovirus T3 Dearing (T3D), the prototypic neurotropic reovirus, causes fatal encephalitis associated with neuronal injury and virus-induced apoptosis throughout the brain. T3D variant K (VarK) is an antigenic variant that has a nearly 1 million-fold reduction in neurovirulence following intracerebral (i.c.) inoculation compared to T3D and a restricted pattern of central nervous system injury with damage limited to the hippocampus, sparing other brain regions. We wished to determine whether the restricted pattern of VarK-induced injury was due to a reduced capacity to replicate in or injure cortical, as opposed to hippocampal, tissue. We found that following i.c. inoculation, VarK grew to similar titers as T3D in the hippocampus but had significantly lower titers in the cortex. Both viruses grew to identical titers and infected the same percentage of cells in mouse primary hippocampal cultures (MHC). In mouse primary cortical cultures (MCC) both the number of infected cells and the viral yield per infected cell were significantly lower for VarK than T3D. VarK-induced apoptosis was limited to the hippocampus in vivo, and in vitro both viruses induced apoptosis equally in MHC but VarK induced significantly less apoptosis than T3D in MCC. Growth of T3D in MCC was reduced to levels comparable to those of VarK following treatment of MCC with caspase inhibitors. Conversely, induction of apoptosis in VarK-infected MCC with fatty acid synthase-activating antibody significantly enhanced viral yield. These results suggest that the decreased neurovirulence of VarK may be due to its failure to efficiently induce apoptosis in cortical neurons. PMID- 15113926 TI - Impact of mutations in the coreceptor binding site on human immunodeficiency virus type 1 fusion, infection, and entry inhibitor sensitivity. AB - An increasingly large number of antiviral agents that prevent entry of human immunodeficiency virus (HIV) into cells are in preclinical and clinical development. The envelope (Env) protein of HIV is the major viral determinant that affects sensitivity to these compounds. To understand how changes in Env can impact entry inhibitor sensitivity, we introduced six mutations into the conserved coreceptor binding site of the R5 HIV-1 strain YU-2 and measured the effect of these changes on CD4 and coreceptor binding, membrane fusion levels and rates, virus infection, and sensitivity to the fusion inhibitors enfuvirtide (T 20) and T-1249, the CCR5 inhibitor TAK-779, and an antibody to CD4. The mutations had little effect on CD4 binding but reduced CCR5 binding to various extents. In general, reductions in coreceptor binding efficiency resulted in slower fusion kinetics and increased sensitivity to TAK-779 and enfuvirtide. In addition, low CCR5 binding usually reduced overall fusion and infection levels. However, one mutation adjacent to the bridging sheet beta21 strand, P438A, had little effect on fusion activity, fusion rate, infectivity, or sensitivity to enfuvirtide or T 1249 despite causing a marked reduction in CCR5 binding and a significant increase in TAK-779 sensitivity. Thus, our findings indicate that changes in the coreceptor binding site of Env can modulate its fusion activity, infectivity, and entry inhibitor sensitivity by multiple mechanisms and suggest that reductions in coreceptor binding do not always result in prolonged fusion kinetics and increased sensitivity to enfuvirtide. PMID- 15113927 TI - Receptor-dependent coronavirus infection of dendritic cells. AB - In several mammalian species, including humans, coronavirus infection can modulate the host immune response. We show a potential role of dendritic cells (DC) in murine coronavirus-induced immune modulation and pathogenesis by demonstrating that the JAW SII DC line and primary DC from BALB/c mice and p/p mice with reduced expression of the murine coronavirus receptor, murine CEACAM1a, are susceptible to murine coronavirus infection by a receptor-dependent pathway. PMID- 15113928 TI - Comparison of the Rta/Orf50 transactivator proteins of gamma-2-herpesviruses. AB - The viral immediate-early transactivator Rta/Orf50 is necessary and sufficient to initiate Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8) reactivation from latently infected cells. Since Rta/Orf50 is conserved among all known gamma-2-herpesviruses, we investigated whether the murine gamma-68 herpesvirus (MHV-68) and rhesus monkey rhadinovirus (RRV) homologs can functionally substitute for KSHV Rta/Orf50. (i) Our comparison of 12 KSHV promoters showed that most responded to all three Rta/Orf50proteins, but three promoters (vGPCR, K8, and gB) responded only to the KSHV Rta/Orf50 transactivator. Overall, the activation of KSHV promoters was higher with KSHV Rta than with the RRV and MHV-68 Rta. (ii) Only the primate Rta/Orf50 homologs were able to interfere with human p53-depedent transcriptional activation. (iii) Transcriptional profiling showed that the KSHV Rta/Orf50 was more efficient than it's homologs in inducing KSHV lytic transcription from the latent state. These results suggest that the core functionality of Rta/Orf50 is conserved and independent of its host, but the human protein has evolved additional, human specific capabilities. PMID- 15113929 TI - Human immunodeficiency virus type 1 envelope glycoproteins that lack cytoplasmic domain cysteines: impact on association with membrane lipid rafts and incorporation onto budding virus particles. AB - The human immunodeficiency virus type 1 (HIV-1) envelope comprises a surface gp120 and a transmembrane gp41. The cytoplasmic domain of gp41 contains cysteine residues (C764 and C837) which are targets for palmitoylation and were reported to be required for envelope association with lipid rafts and assembly on budding virions (I. Rousso, M. B. Mixon, B. K. Chen, and P. S. Kim, Proc. Natl. Acad. Sci. USA 97:13523-13525, 2000). Several infectious HIV-1 clones contain envelopes that have no gp41 cytoplasmic cysteines. Since no other gp41 amino acid is a target for palmitoylation, these clones imply that palmitoylation is not essential for envelope trafficking and assembly. Here, we show that HIV-1 envelope mutants that lack gp41 cytoplasmic cysteines are excluded from light lipid rafts. Envelopes that contained residues with bulky hydrophobic side chains instead of cysteines retained their association with heavy rafts and were nearly fully functional for incorporation into virions and infectivity. Substitution of cysteines with alanines or serines eliminated raft association and more severely reduced envelope incorporation onto virions and their infectivity. Nevertheless, the A764/A837 mutant envelope retained nearly 40% infectivity compared to the wild type, even though this envelope was excluded from lipid rafts. Our results demonstrate that gp41 cytoplasmic cysteines that are targets for palmitoylation and are required for envelope trafficking to classical lipid rafts are not essential for HIV-1 replication. PMID- 15113930 TI - Classical swine fever virus glycoprotein E rns is an endoribonuclease with an unusual base specificity. AB - The glycoprotein E(rns) of pestiviruses is a virion-associated and -secreted RNase that is involved in virulence. The requirements at the cleavage site in heteropolymeric RNA substrates were studied for E(rns). Limited digestion of heteropolymeric RNA substrates indicated a cleavage 5' of uridine residues irrespective of the preceding nucleotide (Np/U). To further study specificity radiolabeled RNA, molecules of 45 to 56 nucleotides in length were synthesized that contained no or a single Np/U cleavage site. Cleavage was only observed in substrates containing an ApU, CpU, GpU, or UpU dinucleotide and occurred in two steps, an initial NpU-specific and a consecutive unspecific degradation. The NpU specific cleavage was resistant to 7 M urea while the second-order cleavage was sensitive to denaturation. Kinetic analyses revealed that E(rns) is a highly active endoribonuclease (k(cat)/K(m) = 2 x 10(6) to 10 x 10(6) M(-1) s(-1)) with a strong affinity to NpU containing single-stranded RNA substrates (K(m) = 85 to 260 nM). PMID- 15113931 TI - Induction of disease by a molecularly cloned highly pathogenic simian immunodeficiency virus/human immunodeficiency virus chimera is multigenic. AB - One of three full-length infectious molecular clones of SHIV(DH12R), designated SHIV(DH12R-CL-7) and obtained from productively infected rhesus monkey peripheral blood mononuclear cells, directed rapid and irreversible loss of CD4+ T cells within 3 weeks of its inoculation into Indian rhesus monkeys. Induction of complete CD4+ T-cell depletion by SHIV(DH12R-CL-7) was found to be dependent on inoculum size. The acquisition of this pathogenic phenotype was accompanied by the introduction of 42 amino acid substitutions into multiple genes of parental nonpathogenic SHIV(DH12). Transfer of the entire SHIV(DH12R-CL-7) env gene into the genetic background of nonpathogenic SHIV(DH12) failed to confer the rapid CD4+ T-lymphocyte-depleting syndrome; similarly, the substitution of gag plus pol sequences from SIV(smE543) for analogous SIV(mac239) genes in SHIV(DH12R-CL-7) attenuated the pathogenic phenotype. Amino acid changes affecting multiple viral genes are necessary, but insufficient by themselves, to confer the prototypically rapid and irreversible CD4+ T-cell-depleting phenotype exhibited by molecularly cloned SHIV(DH12R-CL-7). PMID- 15113932 TI - Effects of antibody on viral kinetics in simian/human immunodeficiency virus infection: implications for vaccination. AB - Passive antibody treatment of macaques prior to simian/human immunodeficiency virus infection produces "sterilizing immunity" in some animals and long-term reductions in viral loads in others. Analysis of viral kinetics suggests that antibody mediates sterilizing immunity by its effects on the initial viral inoculum. By contrast, reduction in peak viral load later in infection prevents CD4 depletion and contributes to long-term viral control. PMID- 15113933 TI - Characterization of restrictions to human immunodeficiency virus type 1 infection of monocytes. AB - Tissue macrophages are an important cellular reservoir for replication of human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus. In vitro, the ability of macrophages to support viral replication is differentiation dependent in that precursor monocytes are refractory to infection. There is, however, no consensus as to the exact point at which infection is restricted in monocytes. We have revisited this issue and have compared the efficiencies of early HIV-1 replication events in monocytes and in differentiated macrophages. Although virus entry in monocytes was comparable to that in differentiated macrophages, synthesis of full-length viral cDNAs was very inefficient. Relative to differentiated macrophages, monocytes contained low levels of dTTP due to low thymidine phosphorylase activity. Exogenous addition of D-thymidine increased dTTP levels to that in differentiated macrophages but did not correct the reverse transcription defect. These results point to a restriction in monocytes that is independent of reverse transcription precursors and suggest that differentiation dependent cellular cofactors of reverse transcription are rate limiting in monocytes. PMID- 15113935 TI - Intracellular copper transport in mammals. AB - Copper is an essential cofactor for approximately a dozen cuproenzymes in which copper is bound to specific amino acid residues in an active site. However, free cuprous ions react readily with hydrogen peroxide to yield the deleterious hydroxyl radical. Therefore, copper homeostasis is regulated very tightly, and unbound copper is extremely low in concentration. Copper imported by the plasma membrane transport protein Ctr1 rapidly binds to intracellular copper chaperone proteins. Atox1 delivers copper to the secretory pathway and docks with either copper-transporting ATPase ATP7B in the liver or ATP7A in other cells. ATP7B directs copper to plasma ceruloplasmin or to biliary excretion in concert with a newly discovered chaperone, Murr1, the protein missing in canine copper toxicosis. ATP7A directs copper within the transgolgi network to the proteins dopamine beta-monooxgenase, peptidylglycine alpha-amidating monooxygenase, lysyl oxidase, and tyrosinase, depending on the cell type. CCS is the copper chaperone for Cu,Zn-superoxide dismutase; it delivers copper in the cytoplasm and intermitochondrial space. Cox17 delivers copper to mitochondria to cytochrome c oxidase via the chaperones Cox11, Sco1, and Sco2. Other copper chaperones may exist and might include metallothionein and amyloid precursor protein (APP). Genetic and nutritional studies have illustrated the essential nature of these copper-binding proteins; alterations in their levels are associated with severe pathology. PMID- 15113934 TI - Conserved sequence motifs for nucleoside triphosphate binding unique to turreted reoviridae members and coltiviruses. PMID- 15113936 TI - Fredrick John Stare (1910-2002). PMID- 15113937 TI - Carl August Baumann (1906-1999). PMID- 15113938 TI - Flavones mitigate tumor necrosis factor-alpha-induced adhesion molecule upregulation in cultured human endothelial cells: role of nuclear factor-kappa B. AB - Flavones have been classified as anti-atherogenic agents that inhibit monocyte adhesion to stimulated endothelium, possibly by blocking induction of cell adhesion molecules (CAM). This anti-atherogenic feature of these flavonoids appears to be related to their chemical structures. Flavones may interfere with key signaling events involved in endothelial cell activation by inflammatory mediators. This study examined the effects of flavones on the induction of CAM and the translocation and DNA binding of nuclear factor-kappa B (NF-kappa B) in TNF-alpha-activated human umbilical vein endothelial cells (HUVEC). The effects of flavones, luteolin and apigenin, on adhesion of THP-1 monocytes to the TNF alpha-activated HUVEC, protein expression and mRNA levels of vascular cell adhesion molecule-1 (VCAM-1), intracellular cell adhesion molecule-1 (ICAM-1) and E-selectin, and nuclear appearance and DNA binding activity of NF-kappa B were determined. Flavanols, flavonols, and flavanones were used for comparison. TNF alpha significantly induced HUVEC protein expression of VCAM-1, ICAM-1, and E selectin with increasing mRNA levels. Luteolin and apigenin inhibited the TNF alpha-induced upregulation of THP-1 adhesion and VCAM-1 expression; these inhibitory effects were dose-dependent. The flavones at doses of > or =25 micromol/L almost completely abolished the increased CAM protein and mRNA regardless of their anti-oxidative activity. With the exception of the flavonol quercetin, flavonoids had no such effect; quercetin substantially attenuated the CAM induction. The flavones inhibited nuclear translocation and DNA binding activity of the NF-kappa B-containing binding site in the promoter region of the CAM genes in TNF-alpha-activated HUVEC. The inhibition of endothelial CAM induction by flavones is mediated by their interference with the NF-kappa B dependent transcription pathway. Thus, the flavones may hamper initial atherosclerotic events involving endothelial CAM induction. PMID- 15113939 TI - Insulin increases the abundance of the growth hormone receptor in liver and adipose tissue of periparturient dairy cows. AB - After parturition, increased growth hormone (GH) secretion is important to preserve the metabolic homeostasis of energy-deficient dairy cows. Elevated plasma GH promotes lipid mobilization from adipose tissue, but paradoxically, is associated with depressed concentration of insulin-like growth factor-I (IGF-I), a growth factor produced in a GH-dependent fashion in liver. Primary factors regulating GH responses of liver and adipose tissue are poorly understood in periparturient dairy cows. Consistent with insulin being such a factor, its plasma concentration declined concomitantly with net energy balance (EB) and with plasma IGF-I in a group of 9 periparturient dairy cows. To test the role of insulin in regulating cellular determinants of GH responsiveness, hyperinsulinemic-euglycemic clamps were performed on 6 dairy cows in late pregnancy (28 d prepartum) before the reductions in EB, insulin, and IGF-I were initiated, and when they were completed in early lactation (10 d postpartum). Infusion of insulin nearly doubled the plasma concentration of IGF-I (P < 0.001) and hepatic levels of IGF-I mRNA during both states (P < 0.05). In liver, these responses were associated with increased abundance of the GH receptor protein (GHR; P < 0.05), whereas the abundance of intracellular mediators of GH actions (JAK2, STAT5, or STAT3) remained unaffected. Insulin also doubled GHR abundance in adipose tissue (P < 0.01), indicating that this effect is not liver specific. These results raise the possibility that insulin regulates the efficiency of GH signaling in liver and adipose tissue of dairy cows by acting as a rheostat of GHR synthesis. PMID- 15113940 TI - Dietary phytoestrogens increase metabolic resistance (cold tolerance) in long chain acyl-CoA dehydrogenase-deficient mice. AB - We evaluated the role of dietary phytoestrogens (PE) in the disease phenotype of cold intolerance that characterizes long-chain acyl-CoA dehydrogenase-deficient (LCAD-/-) mice, a model of inborn errors of mitochondrial fatty acid beta oxidation. Male LCAD-/- mice were fed a standard diet containing endogenous PE, a PE-free diet, or a PE-free diet that was supplemented with genistein (250 microg/g diet). The standard diet did not restore complete cold tolerance, but it provided more resistance (P = 0.004) to cold challenge than the PE-free diet. There was a nonsignificant difference (P < 0.07) between LCAD-/- mice fed the genistein-supplemented diet and those fed the PE-free diet. There were no differences in end-point serum glucose concentrations among the 3 groups. Serum FFA were decreased in LCAD-/- mice fed the standard diet compared with those fed the PE-free diet (P = 0.005) and the diet supplemented with genistein (P < 0.001). Serum triglyceride concentrations were greater (P < 0.05) only in LCAD-/- mice fed the genistein-supplemented diet than those fed the standard diet. These results demonstrate the beneficial effects of dietary PE on metabolic tolerance in LCAD-/- mice. Furthermore, they suggest changes that could improve pediatric formula constituents, especially with regard to management of children with inborn errors of fatty acid oxidation. PMID- 15113941 TI - The human fatty acid synthase gene and de novo lipogenesis are coordinately regulated in human adipose tissue. AB - Despite its potential importance in obesity and related disorders, little is known about regulation of lipogenesis in human adipose tissue. To investigate this area at the molecular and mechanistic levels, we studied lipogenesis and the regulation of 1 of its core enzymes, fatty acid synthase (FAS), in human adipose tissue in response to hormonal and nutritional manipulation. As a paradigm for lipogenic genes, we cloned the upstream region of the human FAS gene, compared its sequence to that of FAS orthologs from other species, and identified important regulatory elements that lie upstream of the FAS coding region. Lipogenesis, as assessed by glucose incorporation into lipids, was increased by insulin and more so by the combination of insulin and dexamethasone (Dex, a potent glucocorticoid analogue). In parallel, FAS expression, activity, and gene transcription rate were also significantly increased by these treatments. We also showed that linoleic acid, a representative PUFA, attenuated the actions of insulin and Dex on fatty acid and lipid synthesis as well as FAS activity and expression. Using reporter assays, we determined that the regions responsible for hormonal regulation of the FAS gene lie in the proximal portion of the gene's 5' flanking region, within which we identified an insulin response element similar to the E-box sequence we identified previously in the rat FAS gene. In summary, we demonstrated that lipogenesis occurs in human adipose tissue and can be induced by insulin, further enhanced by glucocorticoids, and suppressed by PUFA in a hormone-dependent manner. PMID- 15113942 TI - Epigallocatechin-3-gallate, a green tea-derived polyphenol, inhibits IL-1 beta dependent proinflammatory signal transduction in cultured respiratory epithelial cells. AB - Polyphenolic components of green tea, such as epigallocatechin-3-gallate (EGCG), have potent anti-inflammatory properties. We previously showed that EGCG inhibits tumor necrosis factor-alpha (TNF-alpha)-mediated activation of the nuclear factor kappa B (NF-kappa B) pathway, partly through inhibition of I kappa B kinase (IKK). The NF-kappa B pathway may also be activated in response to interleukin-1 beta (IL-1 beta) stimulation through a distinct signal transduction pathway. We therefore hypothesized that EGCG inhibits IL-1 beta-mediated activation of the NF kappa B pathway. Because the gene expression of interleukin-8 (IL-8), the major human neutrophil chemoattractant, is dependent on activation of NF-kappa B, IL-8 gene expression in human lung epithelial (A549) cells treated with human IL-1 beta was used as a model of IL-1 beta signal transduction. The EGCG markedly inhibited IL-1 beta-mediated IL-1 beta receptor-associated kinase (IRAK) degradation and the signaling events downstream from IRAK degradation: IKK activation, I kappa B alpha degradation, and NF-kappa B activation. In addition, EGCG inhibited phosphorylation of the p65 subunit of NF-kappa B. The functional consequence of this inhibition was evident by inhibition of IL-8 gene expression. Therefore, the green tea polyphenol EGCG is a potent inhibitor of IL-1 beta signal transduction in vitro. The proximal mechanisms of this effect involve inhibition of IRAK-dependent signaling and phosphorylation of p65. PMID- 15113943 TI - Specific preservation of biosynthetic responses to insulin in adipose tissue may contribute to hyperleptinemia in insulin-resistant obese mice. AB - Obesity is characterized by whole-body insulin resistance, yet the expression of many insulin-stimulated genes, including leptin, is elevated in obesity. These observations suggest that insulin resistance may depend on tissue type and gene. To address this hypothesis, we examined the regulation of immediate-early gene expression in liver and adipose tissue after injection of insulin and glucose, in lean insulin-sensitive, and in A(y)/a obese insulin-sensitive and obese insulin resistant mice. Expression of hepatic jun-B mRNA was robustly increased after insulin injection in lean insulin-sensitive a/a mice and insulin-sensitive A(y)/a mice. In contrast, induction of hepatic jun-B and c-fos gene expression by insulin was markedly attenuated in obese insulin-resistant mice. Surprisingly, induction of adipose jun-B and c-fos gene expression by insulin was markedly enhanced in obese insulin-resistant mice. Furthermore, the expressions of jun-B and leptin were also enhanced in insulin-resistant mice after injection of glucose. Leptin mRNA was positively correlated with blood glucose levels and jun B mRNA in lean but not insulin-resistant mice. Multiple regression analysis indicated that the correlation between leptin mRNA and jun-B mRNA was significant even after removing the effect of blood glucose, but the correlation between leptin mRNA and glucose was no longer significant after removing the effect of jun-B mRNA. These data suggest that some impairments in biosynthetic responses to insulin are manifest primarily in the liver, leading to hyperinsulinemia and stimulating the expression of some adipose insulin-stimulated genes, including leptin. These studies demonstrate the utility of immediate-early gene expression in the analysis of biosynthetic mechanisms of insulin resistance. PMID- 15113944 TI - A solid dietary fat containing fish oil redistributes lipoprotein subclasses without increasing oxidative stress in men. AB - There is a demand and need for healthy solid dietary fats. However, synthetic fats can be tailored to contain specific physiologic properties. Our goal was to design dietary solid test fats that would be both beneficial to the atherogenic lipid profile and stable against lipid peroxidation. Sixteen men (age 35-75 y) substituted 80 g of their normal dietary fat intake with test fat for two periods of 21 d each in a double-blind, randomized, crossover study. Although solid, both test fats were low in cholesterol-raising SFA. Test fat "F" contained 5 g/100 g long chain (n-3) fatty acids matched by oleic acid in test fat "O." Plasma total triacylglycerol (TAG), VLDL TAG, cholesterol in VLDL, and intermediate density lipoproteins (IDL) were lower (P < 0.05), whereas apolipoprotein (apo) B of the large LDL-2 (d = 1031-1042 g/L) subclass, and cholesterol of HDL(2b) subclass, were higher after intake of F than O fat (P < 0.05). There was no difference in the effect on in vivo oxidation measured as the ratio of plasma isoprostanes F(2) to arachidonic acid and urinary isoprostanes, whereas the vitamin E activity/plasma total lipids ratio was higher after intake of F than O (P = 0.008). In conclusion, a solid dietary fat containing (n-3) PUFA decreased plasma TAG, VLDL, and IDL cholesterol, and redistributed lipoprotein subclasses in LDL and HDL, with a higher concentration of the larger and less atherogenic subfractions. These changes took place without an increase in oxidative stress as measured by in vivo markers. PMID- 15113945 TI - A sodium-rich carbonated mineral water reduces cardiovascular risk in postmenopausal women. AB - This study was designed to investigate the possible beneficial effects of consuming a sodium-rich carbonated mineral water on lipoprotein metabolism and to determine whether consumption of this water influences endothelial dysfunction (ED) in postmenopausal women. Women included in the study were amenorrheic (>1 y), healthy, and not obese (BMI < 30 kg/m(2)). The subjects did not take estrogen replacement therapy; supplements of vitamins, minerals, and phytoestrogens; or other medications known to affect bone and lipid metabolism. The study consisted of 2 intervention periods of 2 mo each, during which women drank 1 L/d of a control mineral water (low mineral content) for 2 mo followed by the carbonated mineral water, rich in sodium, bicarbonate, and chloride, for 2 mo. Body weight, height, and blood pressure were measured, and BMI was calculated. Blood samples were taken from fasting subjects and serum was analyzed for total cholesterol, HDL-cholesterol, LDL-cholesterol, triacylglycerols, apolipoprotein AI, apolipoprotein B, soluble intercellular cell adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), and glucose. Blood pressure levels did not change throughout the study. Carbonated water intake decreased total cholesterol and LDL-cholesterol levels by 6.8% (P = 0.001) and 14.8% (P < 0.0001), respectively, whereas HDL-cholesterol concentration increased by 8.7% (P = 0.018), compared to the control period. Therefore, cardiovascular disease (CVD) risk indexes (total cholesterol/HDL-cholesterol and LDL-cholesterol/HDL cholesterol) were markedly reduced (both P < 0.0001). Soluble ICAM-1 and sVCAM-1 levels decreased by 8.4% (P = 0.007) and 14.8% (P = 0.015), respectively. Fasting serum glucose concentration decreased by 6.7% (P < 0.0001). Triacylglycerol levels did not change. Consumption of this sodium rich carbonated water can play a beneficial role in the prevention of CVD and the metabolic syndrome. PMID- 15113946 TI - Dietary iron intake is positively associated with hemoglobin concentration during infancy but not during the second year of life. AB - Iron status during infancy and early childhood reflects highly dynamic processes, which are affected by both internal and external factors. The regulation of iron metabolism seems to be subjected to developmental changes during infancy, although the exact nature of these changes and their implications are not fully understood. We wanted to explore the association between dietary iron intake and indicators of iron status, and to assess temporal changes in these variables. This was done by secondary analysis of data from a recently conducted dietary intervention trial in which healthy, term, well-nourished infants were randomly assigned to consume iron-fortified infant cereals with regular or low phytate content, or iron-fortified infant formula. Dietary iron intake from 6 to 8 mo and from 9 to 11 mo was associated with hemoglobin (Hb) concentration at 9 mo (r = 0.27, P < 0.001) and 12 mo (r = 0.21, P = 0.001), respectively, but iron intake from 12 to 18 mo was not associated with Hb at 18 mo. In contrast, iron intake from 6 to 11 mo was not associated with serum ferritin (S-Ft) at 9 or 12 mo, whereas iron intake from 12 to 17 mo was positively associated with S-Ft at 18 mo (r = 0.14, P = 0.032). These shifts in associations between dietary iron intake, and Hb and S-Ft, respectively, may be due to developmental changes in the channeling of dietary iron to erythropoiesis relative to storage, in the absence of iron deficiency anemia. These observations should be taken into consideration when evaluating iron nutritional status during infancy and early childhood. PMID- 15113947 TI - Waist circumference is a better predictor than body mass index of coronary heart disease risk in overweight premenopausal women. AB - Waist circumference (WC) has been postulated to have stronger associations with biomarkers of coronary heart disease (CHD) than BMI. In this study, we measured the level of activity by determining steps walked per day and select biomarkers for CHD risk in 80 overweight or obese (BMI = 25-37 kg/m(2)) premenopausal women to evaluate whether these biomarkers are associated with WC or BMI. The plasma biomarkers measured, using samples from women who had fasted for 12 h, were lipids, apolipoproteins (apo), LDL peak diameter, LDL susceptibility to oxidation, glucose, leptin, and insulin. We identified subjects with the metabolic syndrome (11%) and insulin resistance (30%) to further distinguish subjects at increased risk for CHD. Both BMI and WC were positively correlated with insulin (r = 0.376 and 0.384, respectively, P < 0.05) and leptin (r = 0.614 and 0.512, respectively, P < 0.01) and negatively correlated with the number of steps taken per day (r = -0.245 and -0.354, respectively, P < 0.05). In addition, WC had positive correlations with diastolic blood pressure (r = 0.250, P < 0.05), plasma triglycerides (TG) (r = 0.270, P < 0.05), and apo C-III (r = 0.240, P < 0.05). Women with BMI > or = 30 kg/m(2) or WC > 88 cm had significantly higher leptin concentrations than women having a BMI < 30 kg/m(2) or a WC < or = 88 cm; women with WC > 88 cm also had higher diastolic pressure (P < 0.05), and higher plasma TG (P < 0.05) and apo C-III (P < 0.05) concentrations than those with WC < or = 88. In addition, subjects with the higher WC walked an average of 1000 fewer steps per day (P < 0.01). These results suggest that WC is a stronger predictor of CHD risk than BMI and is more closely associated with the level of exercise in premenopausal women. PMID- 15113948 TI - Dephytinization of a complementary food based on wheat and soy increases zinc, but not copper, apparent absorption in adults. AB - Complementary foods based on cereals may contain high amounts of phytic acid, which binds strongly to minerals and trace elements. The objective of the study was to evaluate the effect of dephytinization of a cereal-based complementary food on zinc and copper apparent absorption in adults. A dephytinized complementary food (<0.03 mg phytic acid/g) and one containing the native phytic acid concentration (4 mg/g) were labeled extrinsically with stable isotopes ((70)Zn and (65)Cu). Apparent zinc and copper absorption was based on fecal excretion of nonabsorbed labels in 9 adults, using a crossover design. Stable isotopes were quantified by thermal ionization MS. Apparent fractional zinc absorption was significantly higher (P = 0.005; Student's paired t test) from the dephytinized complementary food (34.6 +/- 8.0%; mean +/- SD) than from the complementary food with native phytic acid concentration (22.8 +/- 8.8%). Apparent fractional copper absorption did not differ (P = 0.167; 19.7 +/- 5.1% dephytinized vs. 23.7 +/- 8.1% native phytic acid). These results clearly demonstrate the beneficial effect of dephytinization of a complementary food on fractional absorption of zinc but not of copper in adults. The long-term nutritional benefits of dephytinization of complementary foods should be evaluated in young children. PMID- 15113949 TI - Supplementation of a diet low in carotenoids with tomato or carrot juice does not affect lipid peroxidation in plasma and feces of healthy men. AB - Antioxidant properties of carotenoids are thought to be at least partly responsible for the protective effects of fruits and vegetables rich in carotenoids against colon cancer. There are large amounts of in vitro data supporting this hypothesis. But there is little known about the antioxidant effects of carotenoid-rich food in vivo particularly in the gastrointestinal tract. In a randomized, crossover trial, healthy men (n = 22) who were consuming a low-carotenoid diet drank 330 mL/d tomato juice or carrot juice for 2 wk. Antioxidant capacity was assessed by the "lag time" of ex vivo LDL oxidation induced by copper and lipid peroxidation as determined by measurements of malondialdehyde (MDA) in plasma and feces using HPLC with fluorescence detection. Although consumption of both carotenoid-rich juices for 2 wk increased the carotenoid level in plasma and feces (P < 0.001), the antioxidant capacity of LDL tended to be increased by only approximately 4.5% (P = 0.08), and lipid peroxidation in the men's plasma and feces was not affected. Thus, processes other than lipid peroxidation could be responsible for the preventive effects of tomatoes and carrots against colon cancer. PMID- 15113950 TI - Processed complementary food does not improve growth or hemoglobin status of rural tanzanian infants from 6-12 months of age in Kilosa district, Tanzania. AB - A double-blind, randomized, placebo-controlled trial was conducted from March 2001 to March 2002 involving 309 infants who received either a processed complementary food (CF) or an unprocessed placebo from 6 to 12 mo of age. The groups were comparable in baseline characteristics. The study took place in Kilosa district, Tanzania. The processed CF contained germinated, autoclaved, and dried finger millet (65.2%), kidney beans (19.1%), roasted-peanuts (8%), and mango puree (7.7%). The same blend, but not processed, served as the placebo. Processing increased iron solubility and energy density without affecting viscosity. Mean length for age, weight for age, hemoglobin, and zinc protoporphyrin at 6 and 12 mo did not differ between the 2 groups. The results show that the processed food did not differ from the unprocessed placebo in improving growth, hemoglobin, and iron status of infants when given under the study conditions. The control group consumed equal amounts of macronutrients, and the higher energy density in this study did not seem to have any benefits. In our study, there was a very intensive follow-up; at every encounter with mothers, giving the required amounts and adding extra lipids was strongly reinforced. Under those conditions, a well-balanced complementary food with additional lipids can meet the energy needs of young children. The reduction in phytates by 34% and improvement in iron solubility to 19% due to processing might not have been enough to compensate for the rather low iron content of the complementary food. PMID- 15113951 TI - Exclusive breast-feeding for 6 months, with iron supplementation, maintains adequate micronutrient status among term, low-birthweight, breast-fed infants in Honduras. AB - There is little information on the risk of micronutrient deficiencies during the period of exclusive breast-feeding. We evaluated this among term, low-birthweight (LBW; 1500-2500 g) infants in Honduras. Mother-infant pairs were recruited in the hospital and assisted with exclusive breast-feeding during the first 4 mo. At 4 mo, infants were randomly assigned to either continue exclusive breast-feeding to 6 mo (EBF; n = 59) or be given iron-fortified complementary foods (rice, chicken, fruits, and vegetables) from 4 to 6 mo while continuing to breast-feed (SF, n = 60). Blood samples were collected at 2, 4, and 6 mo and analyzed for hemoglobin (Hb), hematocrit, plasma ferritin, % transferrin saturation, vitamin A, vitamin B 12, folate, zinc, and erythrocyte folate. Infants with Hb < 100 g/L at 2 or 4 mo were given medicinal iron supplements for 2 mo; the proportion administered iron drops did not differ significantly between groups. There was no significant effect of complementary foods on indices of vitamin A, B-12, folate, or zinc status. Among infants not given medicinal iron at 4-6 mo, iron status was higher in the SF group than the EBF group. In those given medicinal iron at 4-6 mo, iron status was higher in the EBF group, suggesting that complementary foods interfered with iron utilization. About half of the infants were anemic by 2 mo, before the age when complementary foods would be recommended. This supports the recommendation that LBW infants should receive iron supplementation in early infancy. Given that infants given iron supplements did not benefit from complementary foods at 4-6 mo, we conclude that exclusive breast-feeding for 6 mo (with iron supplementation) can be recommended for term, LBW infants. PMID- 15113952 TI - Intermittent iron supplementation regimens are able to maintain safe maternal hemoglobin concentrations during pregnancy in Venezuela. AB - Daily iron supplementation programs for pregnant women recommend amounts of iron that are considered by some to be excessive, and either lower-dose or less frequent iron supplementation regimens have been proposed. A randomized, placebo controlled study was performed to assess and compare the relative effectiveness of a weekly (WS) or twice weekly (TW) iron supplementation schedule in maintaining or achieving hemoglobin (Hb) levels at term considered to carry minimal maternal and fetal risk (90-130 g/L). Pregnant women (n = 116) at wk 10 30 of gestation (63 WS and 53 TW) were enrolled in the study (52 in WS and 44 TW completed the study). Women were randomly allocated to receive a 120-mg oral dose of iron as ferrous sulfate and 0.5 mg of folic acid weekly (n = 52) or 60 mg iron and 0.25 mg folic acid and a placebo twice weekly (n = 44). Hb, hematocrit, serum ferritin, and transferrin saturation were estimated at baseline and at 36-39 wk of gestation. Baseline dietary data and the presence and intensity of intestinal helminthic infections were assessed. The duration of supplementation was 14 +/- 4 wk and the median level of adherence was 60.5%. Hb concentrations improved in women following the TW regimen and in women following WS who had low baseline Hb levels. About 89% of WS women and 95% of TW women maintained Hb levels at term (between 90 g/L and 130 g/L), a range associated with optimal pregnancy outcomes. One woman in the TW group exhibited higher Hb levels that potentially carried perinatal risk (>130 g/L). Intermittent iron and folic acid supplementation may be a valid strategy when used as a preventive intervention in prenatal care settings. PMID- 15113953 TI - Cold-storage affects antioxidant properties of apples in Caco-2 cells. AB - Data on the composition of phenolic antioxidant compounds present in food plants and assessment of their activity are essential for epidemiological explanation of the health benefits of fruit and vegetables. Various factors such as cultivation methods, industrial processing, and storage may affect the final concentrations of phytochemicals in food plants and their eventual bioactivity. This study investigated the influence of commercial cold-storage periods on the antioxidant properties of apples grown either by organic or integrated systems. In both cases, total phenolics and total antioxidant activity decreased only in the first 3 mo and only in apples with skin (P < 0.05), suggesting that cold storage rapidly impoverishes these properties in skin but not in pulp. Assessment of antioxidant bioactivity in vitro, measured in terms of intracellular antioxidant, cytoprotective, and antiproliferative activity in human colon carcinoma (Caco-2) cells (differentiated to normal intestinal epithelia for intracellular antioxidant and cytoprotective effects), showed strong, time-related decreases over 6 mo of cold storage for all 3 parameters (P < 0.01), irrespective of the cultivation system. These findings with integrated and organic apples further support the concept that organic systems of cultivation do not generally provide real health benefits. Moreover, the data from the present study clearly show that factors such as cold storage may affect the antioxidant properties of apples. Epidemiological studies on the cancer-preventive benefits of fruits and vegetables should take into account the cold-storage bias for apples, and possibly for other products. PMID- 15113954 TI - Lymphatic fat absorption varies among rats administered dairy products differing in physiochemical properties. AB - We examined in rats the intestinal absorption of fat from dairy products differing in physiochemical properties. Five dairy products (cream cheese, cream, sour cream, butter, and mixed butter) with minor differences in fatty acid composition were administered by gavage to rats, and lymphatic fat absorption was examined. Absorption was followed for 8 h after administration of 300 mg fat from the dairy products. Administration of cream and sour cream resulted in faster lymphatic fat absorption than cream cheese, butter, and mixed butter, and at 8 h the accumulated absorption of fat was significantly higher. The lymphatic absorption of fat after cream cheese administration was similar to the absorption after butter and mixed butter administration up to the 4-h time point; then it increased to a level between that of rats administered cream or sour cream and butter or mixed butter. Overall, these results demonstrated different lymphatic absorption patterns of fat from dairy products differing in physiochemical properties. Because the fatty acid composition of the dairy products differed only slightly, other factors such as viscosity, type of emulsion, particle size, and likely also protein content may have contributed to the differences in absorption. PMID- 15113955 TI - Total equivalent of reactive chemicals in 142 human food items is highly variable within and between major food groups. AB - Many reactive electrophilic chemicals (e.g., acrylamide and hydrazine) occur in foods, and these could individually or cumulatively contribute to human cancer or other diseases. Glutathione (GSH) reacts with and detoxifies electrophilic compounds and is used physiologically to protect against a broad range of toxic and mutagenic compounds. To elucidate the distribution of reactive chemicals in foods, we added a known amount of GSH to 142 commonly consumed food items and assayed the relative amounts of reactive chemicals in terms of the amount of GSH lost during homogenization and extraction, defined quantitatively in terms of glutathione-reactive units (GRUs). Thirty-four items contained GRUs but no detectable GSH; 53 items contained both GSH and GRUs; 18 items contained no GSH or GRUs; and 37 items contained GSH but no detectable GRUs. Among the food groups, cereals, bread, milk, and milk products had relatively high GRU concentrations and low GSH concentrations; several common beverages also had high GRU concentrations and low GSH concentrations; meats and main course dishes were generally low in GRUs and high in GSH. Fruits and vegetables varied in GRU concentration, but most fresh fruits and vegetables had considerably more GSH than GRUs; exceptions were canned vegetables, which had no GSH or GRUs; fruit drinks, which had moderate levels of GRUs and no GSH; and 3 fruits (blueberries, cherries, and prunes), which had high GRU levels. The results provide a database that can be used with food frequency analyses to evaluate the possible association of health risks with the consumption of foods high in GSH-reactive chemicals. PMID- 15113956 TI - Lactobacillus casei alters hPEPT1-mediated glycylsarcosine uptake in Caco-2 cells. AB - Augmentation of the normal flora of the gastrointestinal tract with probiotic bacteria is currently under investigation as a therapeutic tool for several diseases. However, it is unknown whether probiotic bacteria such as Lactobacillus casei alter the expression and function of intestinal transport proteins such as hPEPT1. The effects of 24 and 48 h incubation of Caco-2 cells with 10(8)/L L. casei on the hPEPT1-mediated uptake rate of 20 micro mol/L [(3)H]glycylsarcosine were examined. Dipeptide uptake did not differ from the control at 24 h (15.9 +/- 2.4 vs. 11.5 +/- 1.4 cm.s(-1).mg protein(-1)); however, a significant increase in uptake occurred after 48 h of L. casei treatment (23.7 +/- 1.5 vs. 12.0 +/- 1.9 cm.s(-1).mg protein(-1); P = 0.005). hPEPT1 involvement was confirmed in experiments using excess substrate. Increased uptake of [(3)H]glycylsarcosine appeared to be the result of the direct interaction of the bacteria with Caco-2 cells because conditioned medium had no effect on dipeptide uptake. hPEPT1 mRNA levels did not differ at any time point. These results show that prolonged incubation of Caco-2 cells leads to increased hPEPT1 activity and that this occurs by a mechanism distinct from increased gene expression. PMID- 15113957 TI - Kupffer cell activity is involved in the hepatoprotective effect of dietary oligofructose in rats with endotoxic shock. AB - In the present study, we tested the hypothesis that dietary oligofructose (FOS) can modulate both the response to an endotoxic shock induced by lipopolysaccharide (LPS) administration and the activity of resident hepatic macrophages, i.e., Kupffer cells. Male Wistar rats (n = 5-9 per group) were fed a standard diet or a diet supplemented with 10 g/100 g FOS for 3 wk. LPS (10 mg/kg) or saline were injected i.p. after dietary treatment. After LPS injection, serum levels of tumor necrosis factor (TNF)-alpha, a proinflammatory cytokine, and prostaglandin E(2) (PGE(2)), an immunosuppressive mediator, were higher in FOS treated rats than in control rats. Alanine aminotransferase (ALT) activity was approximately 50% lower than in controls 24 h after LPS administration in FOS treated rats, suggesting less hepatic injury; this was confirmed through histological analysis. FOS treatment increased the number of large phagocytic Kupffer cells, as assessed by histological examination of the liver after colloidal carbon injection into the portal vein. Precision-cut liver slices (PCLS) from FOS-treated rats released more TNF-alpha and PGE(2) into the incubation medium than PCLS from control rats, independently of LPS challenge in vitro. This would suggest that the higher Kupffer cell phagocytic activity and secretion capacity due to FOS supplementation improve LPS clearance in liver tissue and reduce hepatocyte alterations. This study supports the hypothesis that oligofructose might decrease liver tissue injury after endotoxic shock and sepsis. PMID- 15113958 TI - Folic acid supplementation for 4 weeks affects liver morphology in aged rats. AB - Several countries have approved universal folic acid (FA) fortification to prevent neural tube defects and/or high homocysteine levels; this has led to a chronic intake of FA. Traditionally, the vitamin is considered to be safe and nontoxic, except for the potential masking of vitamin B-12 deficiency. Recent reports from our laboratories showed several effects of high-dose folate supplementation in rats. In this work, we compared the effect of FA on the liver of weanling (3 wk) and aged (18 mo) male rats fed either a diet supplemented with 40 mg FA/kg diet or a control diet (1 mg FA/kg diet) for 4 wk. FA supplementation did not alter serum aspartate aminotransferase, alanine aminotransferase, urea, glucose oxidase, total bilirubin, or uric acid. Routine histological staining as well as immunohistochemistry with proliferating cell nuclear antibody for dividing cells, and cytokeratin-8 against bile ductal cells, showed that aged, supplemented rats had the same number of hepatocytes as both control and supplemented weanling rats, and tended to have more (17%, P = 0.07) hepatocytes than aged, control rats. Moreover, the bile duct cells of aged, control rats proliferated and transformed into cholestatic rosettes at a higher frequency than in aged, supplemented rats. The morphology of the liver in weanling rats was similar in both diet groups, and comparable to the supplemented, aged rats, thus indicating that a high intake of FA improves normal liver morphology in livers of aged rats. PMID- 15113959 TI - Breast cancer risk in premenopausal women is inversely associated with consumption of broccoli, a source of isothiocyanates, but is not modified by GST genotype. AB - The role of vegetable consumption in relation to breast cancer risk is controversial. Anticarcinogenic compounds may be present only in specific vegetables, thereby attenuating findings for total vegetable intake. Cruciferous vegetables contain precursors of isothiocyanates (ITCs), which may be chemopreventive through potent inhibition of phase I, and induction of phase II enzymes, such as glutathione S-transferases (GSTs). We investigated associations between consumption of cruciferous vegetables, sources of ITCs, and breast cancer risk, and potential modification of relations by GSTM1 and GSTT1 genotypes. Cases (n = 740) were Caucasian women with incident breast cancer identified from all major hospitals in Erie and Niagara counties. Community controls (n = 810) were frequency matched to cases by age and county. An in-depth interview including a validated FFQ was administered in person. Odds ratios (ORs) and 95% CIs were used to estimate relative risks. Consumption of cruciferous vegetables, particularly broccoli, was marginally inversely associated with breast cancer risk in premenopausal women [4th quartile OR = 0.6, 95% CI (0.40-1.01), P = 0.058]. Associations were weaker or null among postmenopausal women. No significant effects of GST genotype on risk were observed in either menopausal group. These data indicate that cruciferous vegetables may play an important role in decreasing the risk of premenopausal breast cancer. PMID- 15113960 TI - Dietary depletion of vitamin E and vitamin A inhibits mammary tumor growth and metastasis in transgenic mice. AB - We showed previously that dietary antioxidant depletion enhances tumor reactive oxygen species (ROS) and apoptosis, resulting in a reduction in brain tumor size in the TgT(121) transgenic mouse model, a nonmetastatic tumor model. Here, in a transgenic mouse model of mammary tumorigenesis with defined rates of tumor growth and lung-targeted metastasis, we determined the ability of dietary antioxidant depletion to inhibit tumor growth and metastasis. Compared with control mice fed a standard diet, antioxidant-depleted mice exhibited tumor targeted generation of ROS manifested by increased levels of oxidatively modified DNA/RNA (8- hydroxy-2'-deoxyguanine, 8-hydroxyguanine) and lipid peroxidation (4 hydroxy-2-nonenal) in primary and metastatic tumor foci. In addition to increased tumor-targeted ROS, the number of apoptotic cells was increased approximately 500% (P < 0.01) and terminal dUTP nucleotide DNA end-labeling-positive cells 200% (P < 0.01) in mice fed the antioxidant-depleted diet, whereas the percentage of tumor cells undergoing mitosis was >50% lower than in controls (P < 0.01). The proportional distribution of small (<1.5 cm) and large (> or = 1.5 cm) primary mammary tumors differed. The mice fed the antioxidant-depleted diet had more small primary tumors (P <0.05) and fewer large primary tumors (P < 0.05). Importantly, they also had fewer lung metastatic tumor foci compared with mice fed the control diet (4.5 +/- 1.3 vs. 15.8 +/- 8.5 foci/lung, P < 0.01). These findings may be important in understanding the role of dietary antioxidant vitamins in tumor growth and metastasis. PMID- 15113961 TI - beta-Sitosterol, beta-Sitosterol Glucoside, and a Mixture of beta-Sitosterol and beta-Sitosterol Glucoside Modulate the Growth of Estrogen-Responsive Breast Cancer Cells In Vitro and in Ovariectomized Athymic Mice. AB - We hypothesized that the phytosterols beta-sitosterol (BSS), beta-sitosterol glucoside (BSSG), and Moducare (MC; BSS:BSSG = 99:1) could modulate the growth of estrogen-dependent human breast cancer cells in vitro and in vivo. The present study evaluated the estrogenic and antiestrogenic effects of BSS, BSSG, and MC (0.001 to 150 micromol/L) on the proliferation of Michigan Cancer Foundation 7 (MCF-7) cells in vitro. Both BSS (>1 micromol/L) and MC (>50 micromol/L) increased MCF-7 cell proliferation. Treatment with 150 micro mol/L of BSS and MC increased cell growth by 2.4 and 1.5 times, respectively, compared to the negative control (NC) group. However, BSSG had no effect at the concentrations tested. The effects of dietary BSS, BSSG, and MC on the growth of MCF-7 cells implanted in ovariectomized athymic mice were also evaluated. Estrogenic effects of the phytosterols were evaluated in the NC, BSS, BSSG, and MC treatment groups, and antiestrogenic effects were evaluated in the 17 beta-estradiol (E(2)), E(2) + BSS, E(2) + BSSG, and E(2) + MC treatment groups. Mice were treated with dietary BSS (9.8 g/kg AIN93G diet), BSSG (0.2 g/kg diet), or MC (10.0 g/kg diet) for 11 wk. Dietary BSS, BSSG, and MC did not stimulate MCF-7 tumor growth. However, dietary BSS, BSSG, and MC reduced E(2)-induced MCF-7 tumor growth by 38.9% (P < 0.05), 31.6% (P = 0.08), and 42.13% (P < 0.05), respectively. The dietary phytosterols lowered serum E(2) levels by 35.1, 30.2, and 36.5% in the E(2) + BSS, E(2) + BSSG, and E(2) + MC groups, respectively (P < 0.05), compared to that of the E(2) treatment group. Estrogen-responsive pS2 mRNA expression in tumors did not differ among groups, but expression of the antiapoptotic marker B-cell lymphoma/leukemia-2 (bcl-2) in tumors from the E(2) + MC group was downregulated, compared to that of the E(2) treatment group. In summary, BSS and MC stimulated MCF-7 cell growth in vitro. Although BSSG comprises only 1% of MC, BSSG made MC less estrogenic than BSS alone in vitro. However, dietary BSS and MC protected against E(2)-stimulated MCF-7 tumor growth and lowered circulating E(2) levels. PMID- 15113962 TI - Adrenalectomy does not block the inhibition of mammary carcinogenesis by dietary energy restriction in rats. AB - Dietary energy restriction (DER) has been shown to reproducibly inhibit chemically induced mammary carcinogenesis. The inhibitory activity of DER has been reported to be associated with an increase in circulating corticosterone as well as a decrease in insulin-like growth factor 1 (IGF-1). To determine whether the adrenal glands are required for cancer inhibitory activity, the effects of DER were investigated in adrenalectomized (ADX) rats. Female Sprague-Dawley rats, 29-31 per group, were injected with 0.05 g 1-methyl-1-nitrosourea/kg body wt at 21 d of age, sham operated (SHAM) or bilaterally ADX at 24 d of age, and after 3 d adapted to meal feeding during which rats ate ad libitum (AL) or were restricted to 60% of AL energy intake. ADX resulted in a marked reduction in serum corticosterone in both AL and DER rats. Whereas the carcinogenic response in the mammary gland was not statistically different in SHAM-AL and ADX-AL rats, ADX did not block the cancer inhibitory activity of DER. In fact, cancer inhibitory activity was greatest in ADX-DER rats. Circulating levels of glucose, insulin, IGF-1, and IGF binding protein 3 also were reduced in DER rats. Collectively, these findings indicate that adrenal glands are not required for manifestation of the cancer inhibitory activity of DER. If circulation-borne factors such as corticosterone or IGF-1 are involved in the inhibition of mammary carcinogenesis by DER, IGF-1 is likely to play a greater role than corticosterone. PMID- 15113963 TI - Dietary soy sphingolipids suppress tumorigenesis and gene expression in 1,2 dimethylhydrazine-treated CF1 mice and ApcMin/+ mice. AB - Dietary supplementation with milk sphingolipids inhibits colon tumorigenesis in CF1 mice treated with a colon carcinogen [1,2-dimethylhydrazine (DMH)] and in multiple intestinal neoplasia (Min) mice, which develop intestinal tumors spontaneously. Plant sphingolipids differ structurally from those of mammals [soy glucosylceramide (GlcCer) consists predominantly of a 4,8-sphingadiene backbone and alpha-hydroxy-palmitic acid], which might affect their bioactivity. Soy GlcCer was added to the AIN-76A diet (which contains <0.005% sphingolipid) to investigate whether it would also suppress tumorigenesis in these mouse models. Soy GlcCer reduced colonic cell proliferation in the upper half of the crypts in mice treated with DMH by 50 and 56% (P < 0.05) at 0.025 and 0.1% of the diet (wt/wt), respectively, and reduced the number of aberrant colonic crypt foci (an early marker of colon carcinogenesis) by 38 and 52% (P < 0.05). Min mice fed diets containing 0.025 and 0.1% (wt/wt) soy GlcCer developed 22 and 37% fewer adenomas (P < 0.05), respectively. The effects of dietary sphingolipids on gene expression in the intestinal mucosal cells of Min mice were analyzed using Affymetrix GeneChip microarrays. Soy GlcCer affected the expression of 96 genes by > or = 2-fold in a dose-dependent manner, increasing 32 and decreasing 64. Decreases in the mRNA expression of two transcription factors associated with cancer, hypoxia-induced factor 1 alpha (HIF1 alpha) and transcription factor 4 (TCF4), were confirmed by quantitative RT-PCR. In conclusion, soy GlcCer suppressed colon tumorigenesis in two mouse models; hence, plant sphingolipids warrant further investigation as inhibitors of colon cancer. Because soy contains relatively high amounts of GlcCer, sphingolipids may partially account for the anticancer benefits attributed to soy-based foods. PMID- 15113964 TI - Conjugated eicosapentaenoic acid (EPA) inhibits transplanted tumor growth via membrane lipid peroxidation in nude mice. AB - Both conjugated linoleic acid (CLA) and eicosapentaenoic acid (EPA) have an antitumor effect. Hence, we hypothesized that a combination of conjugated double bonds and an (n-3) highly unsaturated fatty acid would produce stronger bioactivity. To verify the antitumor effect of conjugated EPA (CEPA), we transplanted DLD-1 human colon tumor cells into nude mice, and compared the tumor growth between CEPA-fed mice and CLA- and EPA-fed mice. After tumor cell inoculation, mice were assigned to 1 of 4 groups (control, CLA, EPA, and CEPA) consisting of 10 mice each. The control group received only safflower oil fatty acids, whereas the remaining groups received a mixture of safflower oil fatty acids and 20 g/100 g of total fatty acids as CLA, EPA, or CEPA. Mice were fed once every 2 d for 4 wk at a dose of 50 mg/mouse at each feeding. After 4 wk, tumor growth in CEPA-fed mice was significantly suppressed, compared with that in CLA- (P < 0.005) and EPA-fed mice (P < 0.001). DNA fragmentation in the tumor tissues of the CEPA-fed mice occurred more frequently than in the CLA- (P < 0.001) and EPA-fed mice (P < 0.001), suggesting that CEPA induced apoptosis in the tumor tissues. To further investigate the mechanism, the level of oxidative stress in the tumor tissues was determined. The CEPA-fed mice showed significant lipid peroxidation, compared with the CLA- (P < 0.001) and EPA-fed mice (P < 0.001). Therefore, we verified that CEPA has a stronger in vivo antitumor effect than EPA and CLA, and that CEPA acts through induction of apoptosis via lipid peroxidation. PMID- 15113965 TI - Daily iron supplementation is more efficacious than twice weekly iron supplementation for the treatment of childhood anemia in western Kenya. AB - A recent meta-analysis of 14 clinical trials indicated that daily compared with intermittent iron supplementation resulted in significantly greater hematological improvement in pregnant women. No such definitive beneficial effect was demonstrated in preschool children. We compared the efficacy of daily and twice weekly iron supplementation for 6 wk under supervised and unsupervised conditions in the treatment of mild and moderate anemia [hemoglobin (Hb) 50-109 g/L] in children aged 2-59 mo living in a malaria-endemic area of western Kenya. The study was a cluster-randomized trial using a factorial design; participants were aware of the treatment assigned. All children (n = 1049) were administered a single dose of sulfadoxine-pyrimethamine at enrollment followed by 6 wk of daily supervised iron supplementation [3-6 mg/(kg.d)], twice weekly supervised iron supplementation [6-12 mg/(kg.wk)], daily unsupervised iron supplementation, or twice weekly unsupervised iron supplementation. In the supervised groups, Hb concentrations at 6 and 12 wk (6 wk postsupplementation) were significantly higher in children given iron daily rather than twice weekly [mean (95% CI) difference at 6-wk: 4.2 g/L (2.1, 6.4); 12-wk: 4.4 g/L (1.8, 7.0)]. Among the unsupervised groups, Hb concentrations were not different at 6 wk [mean (95% CI) difference: 0.86 g/L (-1.4, 3.1)], but significantly higher at 12 wk for those assigned daily iron [mean (95% CI) difference: 3.4 g/L (0.79, 6.0), P = 0.02]. In this malarious area and after initial antimalarial treatment, 6 wk of daily iron supplementation results in better hematological responses than twice weekly iron supplementation in the treatment of anemia in preschool children, regardless of whether adherence can be ensured. PMID- 15113966 TI - The challenge of measuring global fruit and vegetable intake. AB - The WHO recently conducted, within its Global Burden of Disease 2000 Study, a Comparative Risk Assessment (CRA) to estimate the global health effect of low fruit and vegetable intake. This paper summarizes the methods used to obtain exposure data for the CRA and provides estimates of worldwide fruit and vegetable intakes. Intakes were derived from 26 national population-based surveys, complemented with food supply statistics. Estimates were stratified by 14 subregions, 8 age groups, and gender. Subregions were categorized on the bases of child mortality under age 5 y and 15- to 59-y-old male mortality (A: very low child and adult mortality; B: low child and adult mortality; C: low child, high adult mortality; D: high child and adult mortality; E: high child, very high adult mortality). Mean intakes were highest in Europe A [median = 449 g/(person.d)] and the Western Pacific Region A. They were lowest in America B [median = 192 g/(person.d)], and low in Europe C, the South East Asian Regions B and D, and Africa E. Children and elderly individuals generally had lower intakes than middle-aged adults. SDs varied considerably by region, gender, and age [overall median = 223 g/(person.d)]. Assessing exposure levels for the CRA had major methodological limitations, particularly due to the lack of nationally representative intake data. The results showed mean intakes generally lower than current recommendations, with large variations among subregions. If the burden of disease attributable to dietary factors is to be assessed more accurately, more countries will have to assess the dietary intake of their populations using comparable methods. PMID- 15113967 TI - Dietary fiber and C-reactive protein: findings from national health and nutrition examination survey data. AB - A higher intake of dietary fiber may decrease the risk of developing cardiovascular disease. We examined the association between dietary fiber and serum concentration of C-reactive protein (CRP), a possible predictor of cardiovascular events, using data from the National Health and Nutrition Examination Survey 1999-2000. Among 3920 participants > or = 20 y old, dietary fiber intake was inversely associated with serum CRP concentration. The odds ratio (OR) for increased CRP concentration (>3.0 mg/L) was 0.49 (95% CI 0.37 0.65; P for trend < 0.001) for the highest quintile of fiber intake compared with the lowest. Adjustment for age, gender, race, education, smoking, physical activity, BMI, total energy, and fat intake resulted in a slight attenuation (OR 0.59; CI 0.41-0.85; P for trend = 0.006). Excluding participants with cardiovascular conditions, diabetes, or cancer did not alter the results. Our findings indicate that fiber intake is independently associated with serum CRP concentration and support the recommendation of a diet with a high fiber content. PMID- 15113968 TI - Adjustments to the modified relative dose response (MRDR) test for assessment of vitamin A status minimize the blood volume used in piglets. AB - The modified relative dose response (MRDR) test is widely used in public health research to assess vitamin A (VA) status of populations and individuals. However, method adjustments intended to make the test more useful in large field studies and/or less invasive have not been systematically verified. To compare the similarity between modified tests and the standard MRDR test, and validate both modified and standard tests against liver reserves of VA, we used a piglet model. Following the typical MRDR procedure, piglets (n = 10) were dosed with 5.3 micromol 3,4-didehydroretinyl acetate. Method adjustments were made to the postdose blood sample collection time to decrease both the amount of serum analyzed and sample throughput time. We collected 3 blood samples/piglet at 3, 5, and 7 h or 4, 6, and 8 h postdose. Postdose blood samples obtained between 4 and 7 h gave MRDR values that did not differ. Serum volumes as small as 200 microL, half the volume of the standard method, yielded accurate MRDR values. Method adjustments to reduce sample throughput time require further investigation. In conclusion, because 200 microL of serum can be used in the test, only 0.5 mL, as opposed to 1 mL of blood has to be collected from an individual. This adjustment allows for easier application of the test to individuals, especially infants, from whom it is difficult to obtain a large venous blood sample, thus increasing the utility of the test for researchers. PMID- 15113969 TI - A combined 13CO2/H2 breath test can be used to assess starch digestion and fermentation in humans. AB - Ingestion of starch from corn (naturally enriched with (13)C) should produce (13)CO(2) after small intestinal digestion and (13)CO(2) and H(2) from colonic fermentation. This study used a combined (13)CO(2)/H(2) breath test to assess the digestion and fermentation of resistant starch and to show that the test could detect changes in digestibility due to cooking. Volunteers consumed 40 g digestible cornstarch with water (n = 8), or 40 g resistant cornstarch in liquid (n = 12) or cooked into a pancake (n = 4). Interval breath sampling was performed and analyzed for (13)CO(2) and H(2). Ingestion of resistant starch produced a double-peaked (13)CO(2) excretion curve. The first increase in (13)CO(2) occurred at the same time as excretion from digestible starch (55 +/- 9 and 68 +/- 9 min, respectively), which was due to small intestinal digestion. The second increase in (13)CO(2) was accompanied by an increase in H(2) excretion (432 +/- 15 and 428 +/- 48 min, respectively), which was indicative of colonic bacterial fermentation. Cooking resistant starch increased its degree of digestion from 36 to 72%. The (13)CO(2)/H(2) breath test can be used to estimate digestion and fermentation of starches in different physiologic and pathologic conditions. PMID- 15113970 TI - A single measurement is inadequate to estimate enterolactone levels in danish postmenopausal women due to large intraindividual variation. AB - Single measurements of enterolactone (ENL) used in epidemiologic studies are influenced by intraindividual variation. The objective of this controlled study was to investigate short-term intraindividual variations in serum and urine ENL. Based on these variations, the number of samples required to describe the basal ENL level was estimated. Healthy Danish postmenopausal women (n = 6) aged 54-67 y completed 3 study periods of 24 h within 2 mo. Blood samples were collected at 0, 4, 6, 8, 12, and 24 h and 24-h urine samples were collected. A low-lignan, standardized diet of 3 meals was served. ENL was measured by time-resolved fluoroimmunoassay. Intraindividual and interindividual variations were estimated using a mixed model with repeated measurements. Significant and systematic intraindividual within-day variations (CV) of 31% were observed in serum. Intraindividual day-to-day variations were 56% and overall intraindividual variation of samples collected at random times and on different days was estimated to be 64%. Describing this overall variation required 7 blood samples when estimated with a precision of 50% and 95% confidence. Day-to-day variations in 24-h urine samples were 49%. Large within-day and day-to-day variations suggest that a single measurement of ENL is inadequate to estimate the basal ENL level. PMID- 15113971 TI - The effect of Na2EDTA on iron absorption from ferrous fumarate. PMID- 15113972 TI - Introduction to and perspectives from the Fifth International Symposium on the Role of Soy in Preventing and Treating Chronic Disease. PMID- 15113973 TI - Phytoestrogens and vitamin D metabolism: a new concept for the prevention and therapy of colorectal, prostate, and mammary carcinomas. AB - Epidemiologic studies suggest that nutritional phytoestrogens contained in soy are causally related to protection against hormone-dependent cancers. The incidence of colorectal cancer is at least 30% lower in women than in men in the United States. This suggests that estrogen and, conceivably, nutritional phytoestrogens are protective compounds against colorectal cancer for both sexes. Prevention of colorectal, mammary, and prostate cancer may also depend on optimal synthesis of the antimitotic prodifferentiating vitamin D hormonal metabolite 1,25-(OH)(2)-cholecalciferol (1,25-D3). Cytochrome-P450-hydroxylases responsible for synthesis (CYP27B1; 25-D3-1 alpha-hydroxylase) and catabolism (CYP24; 1,25-D3 24-hydroxylase) of 1,25-D3 are not only present in the kidney but are also expressed in human colonocytes, prostate cells, and mammary cells. In addition, levels of CYP27B1, vitamin D receptor, and estrogen receptor-beta (the high affinity receptor for phytoestrogens) are enhanced early during human colorectal cancer, which suggests an interactive physiological defense against tumor progression. We demonstrate in human mammary and prostate cells concentration dependent regulation of CYP27B1 and of CYP24 by genistein at 0.05-50 micromol/L. The high concentration of 50 micromol/L is very effective in eliminating CYP24 expression in prostate cancer cells. This high concentration can be achieved in vivo in the prostate by an as-yet-unknown concentrative mechanism. Soy feeding, or more effectively genistein feeding, elevates CYP27B1 and reduces CYP24 expression in the mouse colon. In mice fed low nutritional calcium, CYP24 rises in parallel to enhanced colonic proliferation, and genistein counteracts both. We suggest that nutritional soy or genistein can optimize extrarenal 1,25-D3 synthesis, which could result in growth control and, conceivably, in inhibition of tumor progression. PMID- 15113974 TI - Soy protein allergy: incidence and relative severity. AB - Food allergy is a relatively rare and sometimes violent reaction of the immune system to food proteins. The first report characterizing soy allergy appeared in 1934. The Food and Agriculture Organization of the United Nations includes soy in its list of the 8 most significant food allergens. At least 16 potential soy protein allergens have been identified but their relative clinical significance is unknown. Conversely, soy has a long history of successful use in managing cow's milk allergies in infants. To better predict the utility of soy proteins for controlling food allergy, it is important to understand the relative allergenic reactivity of soy compared with other major food proteins. This can be studied using clinical data, animal models, and biochemical approaches; all show diminished reactivity for soy. Clinical studies using in vitro methods and blinded food challenges have generated substantial information. Study populations include high-risk asymptomatic infants and patients with atopic symptoms, positive food challenges, and specific milk allergies. Generally, these studies show lower allergic reactivity for soy proteins vs. other food allergens. Comparisons of food allergen dose-response relationships for triggering allergic symptoms also demonstrate a higher protein concentration threshold for soy (approximately 100 times), indicating lower allergenic reactivity. Extensive investigations of soy immunological reactivity have also been carried out using animal models. Consistent with clinical results, all of these data show substantially diminished immunological reactivity for soy proteins. Biochemical and immunochemical analyses indicate no striking differences between soy and other food proteins that would explain these unexpected differences in allergenic reactivity. PMID- 15113975 TI - Safety of soy-based infant formulas containing isoflavones: the clinical evidence. AB - Soy protein has been used in infant feeding in the West for nearly 100 y. Soy protein infant formulas have evolved in this interval to become safe and effective alternatives for infants whose nutritional needs are not met with human milk or formulas based on cow's milk. Modern soy formulas meet all nutritional requirements and safety standards of the Infant Formula Act of 1980. They are commonly used in infants with immunoglobulin E-mediated cow's milk allergy (at least 86% effective), lactose intolerance, galactosemia, and as a vegetarian human milk substitute. Largely as a result of research in animal models, concerns have been voiced regarding isoflavones in soy infant formulas in relation to nutritional adequacy, sexual development, neurobehavioral development, immune function, and thyroid disease. We discuss the available clinical evidence regarding each of these issues. Available evidence from adult human and infant populations indicates that dietary isoflavones in soy infant formulas do not adversely affect human growth, development, or reproduction. PMID- 15113976 TI - Soy isoflavones--phytoestrogens and what else? AB - The weak estrogen-like properties of isoflavonoids were discovered over 50 y ago. In recent years, the overall effects of the isoflavones in soy on human health have been the subject of lively debate largely based on their presumed estrogenic properties. Missing from our knowledge base is the systematic identification of the cellular and biochemical targets of isoflavones and the mechanisms that they influence. Because of the benign effects of isoflavones on cellular integrity, a concentration of the isoflavones can be reached in cell culture models where almost any process can be modulated. Modern systems biology approaches and high dimensional analysis techniques offer new ways to better understand the function of how cells and integrated biological mechanisms respond to compounds such as isoflavones. Data from experiments using DNA microarray analysis for examining the effects of genistein in the developing rat uterus indicate that genistein alters the expression of 6-8 times as many genes as does a physiological estrogen such as 17 beta-estradiol. Although these new approaches are exciting, their incipient high dimensionality places considerable strain on the quality of experimental design and meaningful statistical interpretation of the resulting data. In another approach using affinity chromatography methods, DING, a novel genistein-binding protein of yet unknown function, was isolated from human breast cancer MCF-7 cells. Its function remains to be established. PMID- 15113977 TI - Not all soy products are created equal: caution needed in interpretation of research results. AB - Interest in the health benefits of soy foods has been intense among the research community, health professionals, and the public. At the same time, potential concerns associated with soy consumption, especially as related to soy isoflavones, have tempered the enthusiasm for making public health recommendations. On both accounts, the primary soybean isoflavone, genistein, has received the most attention. Because consumers are becoming increasingly confused by the often conflicting dietary messages, a balanced and accurate view of the risks and benefits of soy foods and soy food components is essential. Even among health professionals, confusion exists about proper nomenclature and about the precise composition of the agents under investigation. Levels of isoflavones are frequently assumed to be constant within categories of soy foods, and intakes are estimated rather than being directly analyzed. Furthermore, all too often research dealing singularly with genistein is interpreted by both health professionals and the media as equating directly with soy. Researchers often fail to fully understand the implications of their research outcomes and the context in which those outcomes should be placed. With the hundreds of publications yearly on soy and isoflavones, it is especially important to consider the literature in its entirety when making pronouncements about health effects. Efforts are needed by all to reduce the public confusion by adapting standardized approaches to the reporting of data. This paper provides a framework for both standardization of nomenclature and appropriate interpretation of data. PMID- 15113980 TI - LMWH for perioperative anticoagulation in patients on chronic warfarin therapy. AB - OBJECTIVE: To review current data regarding low-molecular-weight heparin (LMWH) use for perioperative anticoagulation in patients receiving chronic warfarin therapy. DATA SOURCES: Data were obtained from the Sixth American College of Chest Physicians Consensus Conference on Antithrombotic Therapy guidelines and a MEDLINE search (1996-January 2003). Search terms included heparin, low-molecular weight heparin, warfarin, perioperative care, and anticoagulants. DATA SYNTHESIS: Heparin is the most common agent used as bridge therapy for perioperative anticoagulation in patients on chronic warfarin therapy; LMWHs are also used. Studies that evaluated enoxaparin and/or dalteparin were reviewed. CONCLUSIONS: Although published studies demonstrate efficacy and safety of LMWHs, more data are needed to support their use as bridge therapy. PMID- 15113981 TI - Successful treatment of Staphylococcus epidermidis hip prosthesis infection with oral linezolid. AB - OBJECTIVE: To report a case of Staphylococcus epidermidis infection of a hip prosthesis successfully treated with oral linezolid. CASE SUMMARY: A 46-year-old woman developed methicillin-resistant S. epidermidis (MRSE) infection of her prosthetic hip. She received oral linezolid 600 mg twice daily for one month; after that time, the biological inflammatory markers returned to normal. DISCUSSION: One of the most serious complications of arthroplasty is joint prosthesis infection. It is mainly caused by gram-positive bacteria, in particular those of the genus staphylococcus. The increasing prevalence of gram positive cocci that are resistant to antimicrobial agents has complicated the treatment of serious infections. CONCLUSIONS: Oral linezolid appears to be an effective and well-tolerated treatment option for hip prosthesis infections due to MRSE. PMID- 15113982 TI - Rash associated with piperacillin/tazobactam administration in infectious mononucleosis. AB - OBJECTIVE: To report a case of piperacillin/tazobactam-induced rash in a patient with infectious mononucleosis. CASE SUMMARY: A 25-year-old white man developed a rash while receiving piperacillin/tazobactam 3.375 g intravenously every 6 hours and gentamicin for osteomyelitis complicating a left femur fracture secondary to a motorcycle accident. Due to progression of the rash following additional doses of piperacillin/tazobactam during hospitalization, the patient's antimicrobial regimen was changed to vancomycin and meropenem. Subsequently, a mononucleosis spot test was positive, and both Epstein-Barr virus (EBV) immunoglobulin (Ig) G and IgM antibodies were positive. The rash rapidly resolved with the discontinuation of piperacillin/tazobactam. DISCUSSION: Although the development of rash following the administration of several different antimicrobials, especially ampicillin, has been previously reported, this is the first report of piperacillin/tazobactam-induced rash in infectious mononucleosis. The rash is generally self-limiting and usually resolves within days of discontinuing the causative antimicrobial agent. An altered drug metabolism or an immune-mediated process has been suggested as the potential mechanism for rash development. CONCLUSIONS: Prior reports of antimicrobial-induced rash in infectious mononucleosis and a positive laboratory diagnosis of EBV in our patient with no history of penicillin allergy support the identification of piperacillin/tazobactam as the inducer of the rash. According to the Naranjo probability scale, the association of piperacillin/tazobactam with the rash was classified as probable. PMID- 15113983 TI - Pharmacist and physician collaboration in the patient's home. AB - OBJECTIVE: To review the historical perspective and current incidence of home visits by physicians and pharmacists, discuss the concept of pharmacists providing care "incident-to" the physician, and introduce a new endeavor of collaborative, incident-to home visits by a pharmacist and physician. DATA SOURCES: A literature search was performed using MEDLINE (1966-May 2003). The search terms used were home visit, house call, incident-to, collaborative care, physician, and pharmacist. STUDY SELECTION AND DATA EXTRACTION: All pertinent articles were evaluated. DATA SYNTHESIS: Home visits were once considered a hallmark of patient care, but have decreased to an average of 0.4 home visits per week for primary care physicians. Home visits are essential for patients unable to commute to the office due to a disability or illness. However, home visits also allow the physician to determine how conditions affect a patient's everyday life, in addition to assessing the living environment. Minimal documented evidence exists of pharmacists making home visits and no documented evidence exists of pharmacists and physicians collaboratively making home visits. The pharmacist complements the value of the visit by assessing medication appropriateness as well as medication administration, storage, accessibility, and adherence. CONCLUSIONS: Current practice styles often compromise services that were once paramount to patient care, as evidenced by a decline in home visits. Recent trends in health care attempt to compensate for high-volume practices and managed care restrictions. A collaborative relationship between a pharmacist and physician with visits into the patient's home is a movement toward improved continuity of care. PMID- 15113984 TI - Evaluation of community pharmacists' recommendations to standardized patient scenarios. AB - BACKGROUND: Recent trends in the global nonprescription drug market show a massive increase in medicine availability to the general public. This places greater responsibility on community pharmacists to ensure that patients exercise self-care appropriately. This small study examined pharmacist performance in facilitating patient self-care. OBJECTIVE: To determine whether an appropriate course of action was taken by UK community pharmacists practicing in one geographic area when presented with 1 of 2 case scenarios: headache or abdominal pain. METHODS: A covert researcher, posing as a patient, spoke with the pharmacist on duty at each of 30 pharmacies, asking for advice on one of the scenarios. The pharmacist was interviewed and their responses were examined by an expert panel. RESULTS: Thirty pharmacies were visited resulting in 28 consultations, 14 each for both scenarios. Pharmacists performed better when counseling the "patient" with abdominal pain than headache. The majority of questions asked were categorized as being relevant by the panel (66% for headache, 89% for abdominal pain), although in both scenarios, the expected outcome of referral was observed only in 7 of the consultations for headache and in 8 cases for abdominal pain. Questioning centered on quantification and clarification of the presenting problem and rarely on questions related to history taking. CONCLUSIONS: Performance levels of the community pharmacists varied considerably. For all pharmacists to perform well, greater emphasis should be placed on eliciting information from the patient to ensure that the appropriate course of action is taken. PMID- 15113985 TI - Montelukast-induced generalized urticaria. AB - OBJECTIVE: To report a case of generalized urticaria induced by montelukast treatment. CASE SUMMARY: A 28-year-old man with allergic rhinitis and moderate persistent asthma developed generalized urticaria 5 days after the initiation of montelukast and inhaled fluticasone. Symptoms disappeared within one day after suspension of both drugs. Two months later, after the resumption of montelukast and fluticasone, the patient developed generalized urticaria and eyelid angioedema, which were successfully treated with intravenous betamethasone, achieving complete remission within hours. After 2 days, the patient resumed inhaled fluticasone only and continued this therapy for several months without any adverse reaction. DISCUSSION: We attributed the adverse reaction to montelukast because of the temporal relationship between use of montelukast and urticaria, the absence of other identified causative factors and other explanations for allergic reactions, and the positive dechallenge and rechallenge. The Naranjo probability scale showed a probable relationship between skin manifestations and montelukast treatment. CONCLUSIONS: The use of antileukotrienes is increasing in asthma therapy. In cases of generalized urticaria in asthmatic patients undergoing montelukast therapy, physicians should be aware of a potential adverse reaction to this drug. PMID- 15113986 TI - Recreational use of sildenafil by HIV-positive and -negative homosexual/bisexual males. AB - OBJECTIVE: To conduct an epidemiologic review of sildenafil in homosexual and bisexual males focusing on concurrent use with club drugs and/or antiretroviral medications. DATA SOURCES: A search of MEDLINE was conducted (1966-September 2003), and an extensive manual review of journals was performed using the key search terms club drugs, sildenafil, antiretrovirals, and HIV. STUDY SELECTION AND DATA EXTRACTION: All articles identified from the data sources were evaluated and information deemed relevant was included. DATA SYNTHESIS: Several epidemiologic studies have reported that sildenafil is abused in a recreational fashion, typically with agents commonly known to be "club drugs." In this setting, sildenafil may reverse the impotence-inducing effects of the club drugs and restore sexual capabilities. Many implications of both recreational and nonrecreational use of sildenafil exist in relation to HIV disease. Concern has been raised regarding the potential effects on high-risk sexual practices. Also, several researchers have documented interactions between sildenafil and various club drugs, as well as with antiretrovirals intended for HIV disease. CONCLUSIONS: Serious concerns exist regarding the concurrent use of sildenafil with antiretrovirals and/or club drugs. Clinicians prescribing sildenafil to patients receiving protease inhibitor-containing antiretroviral regimens should be aware of the potential for reduced sildenafil metabolism with resultant adverse effects. Additionally, untoward effects may also occur when sildenafil is used concurrently with various club drugs including amyl and butyl nitrites ("poppers"). PMID- 15113987 TI - Antimicrobial-associated renal tubular acidosis. AB - OBJECTIVE: To review the literature documenting the association of various antimicrobial medications with the development of renal tubular acidosis (RTA). DATA SOURCES: A search of the English literature via MEDLINE (1966-November 2003) and International Pharmaceutical Abstracts (1970-November 2003) was conducted to identify human reports of RTA associated with various drugs from all available classes of antimicrobial agents. Major search terms included renal tubular acidosis, acidosis, antibiotics, and antimicrobials. Bibliographies of selected articles were also searched to identify additional reports of RTA. STUDY SELECTION AND DATA EXTRACTION: Case reports, observational studies, and experimental studies documenting the association of any antimicrobial agent with the development of RTA were included. DATA SYNTHESIS: Antimicrobial-associated RTA is a relatively uncommon adverse effect, with most reports involving amphotericin B, trimethoprim/sulfamethoxazole, and outdated tetracycline. These agents may induce RTA either through direct tubular toxicity or as a function of their pharmacologic action. The time course for the development of RTA varies depending on the antimicrobial utilized. In most instances, RTA is reversible; however, some patients may experience prolonged recovery after the offending agent is removed. CONCLUSIONS: Given that antimicrobial-associated RTA is a relatively uncommon adverse effect, review of the patient's drug regimen may reveal these agents as otherwise unrecognized causes of RTA. Likewise, underlying causes of RTA other than medications must be ruled out. Diagnosing antimicrobial induced RTA may be difficult, given many of these agents may be used in combination and some are intrinsically nephrotoxic. PMID- 15113988 TI - Postoperative opiate analgesia requirements of smokers and nonsmokers. AB - BACKGROUND: Smoking cigarettes and other forms of nicotine administration appear to blunt the perception of pain. Abrupt discontinuation of nicotine in nicotine dependent patients appears to increase the perception of pain. The clinical importance of nicotine's effect on pain perception is not fully understood. OBJECTIVE: To determine whether smokers who abruptly discontinue smoking as a result of being hospitalized for coronary artery bypass graft (CABG) require more postoperative opiate analgesics than nonsmokers. METHODS: A retrospective review of patients who underwent a CABG was performed. Smokers (n = 20) were compared with nonsmokers (n = 69) with regard to opiate analgesic use during the first 48 hours postoperatively. The use of nonopiate sedatives was also compared between the groups. RESULTS: When normalized for weight and body mass index, smokers required 23% and 33%, respectively, more opiate analgesics than did nonsmokers (p = 0.027 and 0.023, respectively). The percentage of patients who received benzodiazepines postoperatively was similar in the 2 groups. CONCLUSIONS: In this study, smokers deprived of nicotine required a greater amount of opiates in the first 48 hours after CABG than did nonsmokers. Healthcare providers need to be aware of the potential for increased narcotic requirements among nicotine deprived smokers. Further study is needed to determine whether nicotine replacement lessens the requirement for postoperative analgesics in smokers. PMID- 15113989 TI - Cost-effectiveness of escitalopram versus citalopram in the treatment of severe depression. AB - BACKGROUND: Severe depression is associated with an extensive economic burden on both the patient and society. OBJECTIVE: To estimate the cost-effectiveness in Austria of escitalopram compared with citalopram in the management of severe depression (Montgomery-Asberg Depression Rating Scale score > or =30). METHODS: A decision model incorporated treatment paths and associated direct resource use (psychiatric hospitalization, medications, general practitioner and psychiatrist visits, treatment discontinuation, suicide attempts) associated with managing severe depression and the indirect cost of work absenteeism over a 6-month period. Main outcomes were clinical success (remission at 6 mo) and cost (2002 Euros equals approximately 1.25 US) of treatment. The analysis was performed from the Austrian societal and Social Healthcare Insurance System (SHIS) perspectives. Clinical input data were derived from a meta-analysis of 8-week randomized clinical trials. Costs were derived from standard Austrian price lists or from the literature. RESULTS: Six months after the start of treatment, the overall clinical success remission rate was higher for escitalopram (53.7%) than for citalopram (48.7%). From the SHIS perspective, the total expected cost per successfully treated severely depressed patient was 924 (32.1%) lower for escitalopram (2879) compared with citalopram (3803). From the societal perspective, the total expected cost per successfully treated severely depressed patient was 1369 (24.4%) lower for escitalopram (5610) than for citalopram (6979). Sensitivity analyses demonstrated that the model was robust and that, even if citalopram had no acquisition cost, escitalopram remained the dominant strategy for both perspectives. CONCLUSIONS: Treatment with escitalopram was the dominant strategy. These data suggest that escitalopram is a cost-effective antidepressant compared with citalopram in the management of severe depression in Austria. PMID- 15113990 TI - Anti-thrombin III in the management of hematopoietic stem-cell transplantation associated toxicity. AB - OBJECTIVE: To describe the evidence assessing the use of anti-thrombin III (AT III) in the management of toxicity associated with hematopoietic stem-cell transplantation (HSCT)-conditioning regimens. DATA SOURCES: Clinical literature was accessed through conference proceedings, EMBASE, the Cochrane database, and MEDLINE (1966-December 2003). STUDY SELECTION AND DATA EXTRACTION: Case reports, small case series, case-control and cohort studies, and randomized controlled trials of AT-III in HSCT were evaluated. Publications examining AT-III use in the non-HSCT setting were also explored. Key search terms included AT-III, transplantation, and veno-occlusive disease (VOD). DATA SYNTHESIS: Severe VOD and ensuing multiple organ dysfunction is associated with high mortality in HSCT. A low AT-III level has been shown to correlate with the development of organ dysfunction. Phase II trials, case series, and one small, randomized, placebo controlled study suggest a benefit when AT-III therapy is instituted early in the course of VOD/multiple organ dysfunction syndrome. In all of these reports, AT III use was devoid of adverse events. CONCLUSIONS: Although further studies are needed to ascertain the optimal target level, method, and duration of administration, AT-III is still a viable alternative for the treatment of severe VOD and ensuing multiple organ dysfunction. PMID- 15113991 TI - Using the Health Assessment Questionnaire and welfare benefits advice to help people disabled through arthritis to access financial support. AB - OBJECTIVES: To test, in a variety of health settings, the ability of the Health Assessment Questionnaire (HAQ) disability index to predict the eligibility of patients with moderate or severe arthritis for disability living allowance or attendance allowance. METHODS: The study included patients from 20 general practices and four hospital out-patient departments across four areas in the southwest of England. Adults with an established diagnosis of rheumatoid arthritis, or osteoarthritis of the hip or knee, and who were not in receipt of Disability Living Allowance (DLA) or Attendance Allowance (AA) were sent an HAQ. Those who scored 1.5 or more were offered an appointment with a welfare advice worker at which they completed an application for DLA or AA. After 3 months they were contacted by the advice worker and asked about the outcome of their applications. RESULTS: Over half of those who completed an HAQ scored 1.5 or over (moderate to severe disability as measured by the HAQ) and were offered advice from experienced welfare benefits advisors. Of these, 87% applied for DLA or AA. Sixty-nine per cent of the applicants were successful. Those scoring 1.75 and over were more likely to be awarded benefit (73% success CLs 67, 79) than people scoring between 1.5 and 1.625 where 55% (CLs 41,69) of applicants were successful. CONCLUSION: The HAQ was shown to be a good predictor of eligibility for AA or DLA. It can be used, in a variety of health settings, to indicate patients who, with help from an experienced advisor, are likely to gain increased financial help. PMID- 15113992 TI - Ultrasonography of salivary glands in primary Sjogren's syndrome. A comparison with magnetic resonance imaging and magnetic resonance sialography of parotid glands. AB - OBJECTIVES: To evaluate ultrasonography (US) of salivary glands in primary Sjogren's syndrome (SS) and to compare US with parotid magnetic resonance (MR) imaging and MR sialography. METHODS: US examination of parotid, submandibular and sublingual glands was performed on 27 patients with primary SS, 27 healthy controls and 27 symptomatic controls without SS. The results were compared with parotid MR imaging and MR sialography and the clinical features of the patients. RESULTS: Salivary gland abnormalities, parenchymal inhomogeneity or adipose degeneration, were visualized in 21 (78%) SS patients, in one healthy control and in two symptomatic controls by US. Eighteen (67%) patients had changes in the parotid and submandibular glands and 8 (30%) changes in the sublingual glands. In the comparison, MR sialography was found to be the most sensitive method (96%), followed by MR imaging (81%) and US (78%), in detecting glandular changes. The specificity of US was 94%. The US and MR results were related to anti-Ro/SSA positivity but not to saliva secretion. The focus scores were related only to parotid MR imaging findings. CONCLUSIONS: US, MR imaging and MR sialography with modern technology have reached such a good accuracy in visualizing glandular structural changes that they are promising alternatives to the conventional invasive examinations in the diagnostics of SS. PMID- 15113993 TI - Supplementing a home exercise programme with a class-based exercise programme is more effective than home exercise alone in the treatment of knee osteoarthritis. AB - OBJECTIVE: The study aimed to compare the relative effectiveness of providing a home-based exercise programme versus home-based exercise supplemented with an 8 week class-based exercise programme in reducing pain and improving function in patients with knee osteoarthritis. METHODS: Patients (n = 214) with radiologically confirmed knee osteoarthritis were selected. Patients were randomly allocated to either home or home supplemented with class-based exercise programmes. Both groups were given a home exercise programme whilst the supplemented group also attended for 8 weeks of twice weekly knee classes. Assessments of locomotor function, walking pain and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores were made. Assessments were made pre- and post-treatment and also at 6- and 12-month follow-ups. Statistical analysis involved the use of a longitudinal linear model ANCOVA with baseline values entered as a covariate. RESULTS: Patients from the class-based group demonstrated significantly greater improvement in locomotor function (-3.7 seconds; 95% C.I. -4.9 to -2.5) and decrease in walking pain (-15 mm; 95% C.I. 20 to -11) than the home-based group, at 12-months follow-up. CONCLUSIONS: The supplementation of a home based exercise programme with a class-based exercise programme led to clinically significant superior improvement. These improvements were still evident at 12-month review. This is the first trial to evaluate this common physiotherapeutic practice, and based on this evidence, supplementation of home exercises with a class-based exercise programme can be recommended to patients, clinicians and service providers. PMID- 15113994 TI - Rheumatology training in the United Kingdom: the trainees' perspective. AB - BACKGROUND: Rheumatology training has undergone significant changes in the last decade with Calmanization, implementation of the New Deal for junior doctors and newer educational strategies for improving musculoskeletal training, like a core curriculum. However, concerns have been expressed about the quality of postgraduate training programmes in the UK. OBJECTIVES: First, to assess current trainees' perceptions of the quality of core and subspecialty training, the impact of workload on training, and to explore demographic variations in training experience. Secondly, to identify educational strategies that trainees felt would enhance their training. METHODS: The questionnaire was initially distributed to all specialist registrars attending the BSR Annual Meeting in Brighton in April 2002. Subsequently, the questionnaire was posted to all registrars on the Joint Committee for Higher Medical Training list with a reminder after 4 weeks. RESULTS: Trainees rated positively training in routine patient care, musculoskeletal examination and injection skills while training in primary care rheumatology, epidemiology, paediatric rheumatology and sports medicine was rated negatively. There is agreement that the reduction in junior doctors' hours has adversely affected training, and issues relating to workload have overtaken training issues. Trainees undertaking dual accreditation are more likely to feel this. Educational strategies deemed to enhance training included training workshops focused on specific topics, such as musculoskeletal radiology (89.2%), and an adequate debriefing session after an out-patient clinic (81.6%). An independently administered, reliable and valid scale for quality of training could be used to assess regional variations in training and monitoring quality. CONCLUSIONS: The changes to junior doctors' hours, the working patterns of doctors and service commitments have all affected the quality and time available for certain aspects of rheumatological training. A major effort to enhance quality is necessary to ensure that the objectives of training are met within the intended training budget. PMID- 15113995 TI - HLA-B60 and B61 are strongly associated with ankylosing spondylitis in HLA-B27 negative Taiwan Chinese patients. AB - OBJECTIVES: Carriage of HLA-B60 has been shown to increase the risk of ankylosing spondylitis (AS) in B27-positive Caucasian patients, but the association in B27 negative cases is less certain. This study assessed HLA class I gene associations in Chinese HLA-B27-negative AS patients. METHODS: Forty-one Chinese HLA-B27 negative AS patients fulfilling the modified New York diagnostic criteria for AS were recruited, and 11 383 HLA-B27-negative blood donors were used for comparison. HLA-A and -B typing was done with the microlymphocytotoxicity assay. RESULTS: Among the B27-negative AS patients, 21 were male and 20 were female. Of HLA-B alleles, only B60 and B61 significantly increased susceptibility to AS in HLA-B27-negative patients (P<0.001). CONCLUSIONS: In Taiwan Chinese, carriage of B60 is increased in HLA-B27-negative AS patients. The association between B61 and HLA-B27-negative AS patients has not been reported previously. Whether the gene involved is HLA-B60 or B61 or another gene in linkage disequilibrium with these genes is unknown. PMID- 15113996 TI - Perceptions of the risks and benefits of medicines in patients with rheumatoid arthritis and other painful musculoskeletal conditions. AB - OBJECTIVES: To examine beliefs about medication risks and benefits in patients attending a specialist rheumatology clinic for pain-related conditions. METHODS: Eighty-one patients (37 first attendees and 44 existing clinic patients) completed a written questionnaire which asked about current treatments, perceived effectiveness, main risks and benefits, and compliance. RESULTS: Existing clinic patients perceived medications to be more effective and more risky than did the new patients, although both groups rated risks to be moderately low. The main perceived risks were adverse side-effects, although patients reported only moderately low levels of experiencing such effects. CONCLUSIONS: In contrast to some other studies, many of our patients were aware of medication risks and were prepared to accept them provided benefits were seen to be high. Existing clinic patients were more aware of risks and benefits, and reported higher compliance levels than new patients, possibly as a result of the hospital education programme. Future studies should evaluate the effects of the programme more systematically. PMID- 15113997 TI - Chondrogenesis of expanded adult human articular chondrocytes is enhanced by specific prostaglandins. AB - OBJECTIVE: To investigate the effects of the cyclooxygenase-2 (cox-2)-dependent prostaglandins D(2) (PGD(2)), E(2) (PGE(2)) and F(2)alpha (PGF(2)alpha) on the redifferentiation and cartilage matrix production of dedifferentiated articular chondrocytes. METHODS: Human articular chondrocytes from three adult donors were dedifferentiated by monolayer expansion and induced to redifferentiate by culture as 3D pellets in a defined serum-free medium containing TGF-beta(1) and dexamethasone, without or with further supplementation with PGD(2), PGE(2) or PGF(2)alpha. After 2 weeks, pellets were assessed histologically, immunohistochemically, biochemically and by real-time quantitative reverse transcriptase-polymerase chain reaction. RESULTS: All three PGs, but predominantly PGE(2), reduced the staining intensity of pellets for collagen type I, whereas PGD(2) and PGF(2)alpha increased the staining intensity of pellets for collagen type II and glycosaminoglycans (GAG). The GAG/DNA content of pellets was not affected by PGE(2) but was increased 1.5- and 2.1-fold by PGD(2) and PGF(2)alpha respectively. PGE(2) reduced the expression of collagen type I mRNA (9.0-fold), whereas PGD(2) and PGF(2)alpha increased the mRNA expression of collagen type II (6.2- and 4.1-fold respectively) and aggrecan (29.8- and 10.7 fold respectively). CONCLUSION: In contrast to PGE(2), PGD(2) and PGF(2)alpha enhanced chondrogenic differentiation and hyaline cartilage matrix deposition by expanded human articular chondrocytes, and could thus be used to improve in vitro or in vivo cartilage regeneration approaches based on these cells. PMID- 15113998 TI - Extended report: nail disease in psoriatic arthritis--clinically important, potentially treatable and often overlooked. AB - OBJECTIVES: To examine the relationship between the severity of nail disease and characteristics of psoriatic arthritis (PsA). We also wished to assess the clinical management of nail disease in patients with PsA. METHODS: We studied 69 patients with PsA at two visits. On the first visit, a rheumatology assessment of joint, skin and nail disease was made. On the second visit, a detailed dermatology assessment of skin and nails was made. Nail disease was analysed using a 20-nail psoriasis nail severity score (PNSS). RESULTS: There were 57 (83%) patients with clinical evidence of psoriatic nail disease. Although 66 (96%) patients had been treated for skin disease, only one (1%) had received any treatment for nail disease. Severe nail disease measured by the PNSS correlated with severe skin psoriasis as indicated by the percentage of body surface area affected by psoriasis (r = 0.34, P = 0.004) and physician global assessment of psoriasis (r = 0.45, P<0.001). Patients with distal interphalangeal (DIP) joint disease had higher PNSS scores (P = 0.03). The PNSS was also associated with unremitting and progressive arthritis (P<0.001), and correlated with Stanford health assessment questionnaire (HAQ) (r = 0.34, P = 0.004), depression (r = 0.39, P<0.001) and anxiety (r = 0.34, P = 0.004) scores. Compared with dermatology assessment, the rheumatology examination of nail disease had a positive predictive value of 84% and negative predictive value of 83%. CONCLUSIONS: In patients with PsA, the severity of nail disease correlates with indicators of severity of both skin and joint disease. Although rheumatologists can adequately screen for nail disease, the management of this aspect of PsA is often overlooked. PMID- 15113999 TI - Benefit of very early referral and very early therapy with disease-modifying anti rheumatic drugs in patients with early rheumatoid arthritis. AB - OBJECTIVE: Delay of disease-modifying anti-rheumatic drug (DMARD) therapy is a major contributing factor for poor outcome in rheumatoid arthritis (RA). Although early therapy has been shown to be particularly effective, there is still uncertainty about the optimal time point of DMARD introduction. We wanted to test if a therapeutic window of opportunity may exist within the first few months of the disease. METHODS: In this case-control parallel-group study, 20 very early RA (VERA) patients with median disease duration of 3 months were age and gender matched to a group of 20 late early RA (LERA) patients with median disease duration of 12 months until first DMARD initiation. Follow-up time was 36 months. Primary outcome measures were the disease activity score (DAS28) and radiological joint destruction using the Larsen method. RESULTS: Already after 3 months of DMARD therapy we found a significant difference of improvement in favour of the VERA patients in the DAS28. This trend continued over the study period. At study end the DAS28 showed an improvement of 2.8+/-1.5 in the VERA vs 1.7+/-1.2 in the LERA group (P(c)<0.05). The Larsen scores showed a statistically significant retardation of progression in the VERA compared with the LERA. CONCLUSION: Our results indicate that there is a window of opportunity for highly successful treatment of RA in the first year, and especially within the first 3 months of therapy. Thus, early diagnosis and therapy may be the crucial step in achieving optimal control of disease progression and prognosis in RA. PMID- 15114000 TI - Comprehensive transthoracic cardiac imaging in mice using ultrasound biomicroscopy with anatomical confirmation by magnetic resonance imaging. AB - High-frequency ultrasound biomicroscopy (UBM) has recently emerged as a high resolution means of phenotyping genetically altered mice and has great potential to evaluate the cardiac morphology and hemodynamics of mouse mutants. However, there is no standard procedure of in vivo transthoracic cardiac imaging using UBM to comprehensively phenotype the adult mice. In this paper, the characteristic mouse thoracic anatomy is elucidated using magnetic resonance (MR) imaging on fixed mice. Besides the left parasternal and apical windows commonly used for transthoracic ultrasound cardiac imaging, a very useful right parasternal window is found. We present strategies for optimal visualization using UBM of key cardiac structures including: 1) the right atrial inflow channels such as the right superior vena cava; 2) the right ventricular inflow tract via the tricuspid orifice; 3) the right ventricular outflow tract to the main pulmonary artery; 4) the left atrial inflow channel, e.g., pulmonary vein; 5) the left ventricular inflow tract via the mitral orifice; 6) the left ventricular outflow tract to the ascending aorta; 7) the left coronary artery; and 8) the aortic arch and associated branches. Two-dimensional ultrasound images of these cardiac regions are correlated to similar sections in the three-dimensional MR data set to verify anatomical details of the in vivo UBM imaging. Dimensions of the left ventricle and ascending aorta are measured by M-mode. Flow velocities are recorded using Doppler at six representative intracardiac locations: right superior vena cava, tricuspid orifice, main pulmonary artery, pulmonary vein, mitral orifice, and ascending aorta. The methodologies and baseline measurements of inbred mice provide a useful guide for investigators applying the high-frequency ultrasound imaging to mouse cardiac phenotyping. PMID- 15114001 TI - Expression profiling analysis of the metabolic and inflammatory changes following burn injury in rats. AB - Burn injury initiates an inflammatory response as part of the healing process that is associated with extensive metabolic adjustments. While most studies have focused on understanding these changes from a biochemical perspective, not much work has been done to characterize these processes at the gene expression level. As a first step, we have comprehensively analyzed changes in gene expression in rat livers during the first 24 h after burn injury using Affymetrix GeneChips, which showed 339 genes to be differentially expressed at a statistical significance of P < 0.05 and changed at least twofold. Functional classification based on gene ontology terms indicated that two categories, metabolism (28%) and inflammation (14%), accounted for nearly 42%. Detailed analysis of the metabolism group of genes indicated that fatty acid (FA) and triglyceride (TG) biosynthesis in the liver were unchanged, whereas TG utilization, FA import, and beta oxidation increased after burn injury. The increased FA pools after burn injury appear to serve as substrates for ATP production. Following burn injury, the cholesterol biosynthetic pathway was suppressed while cholesterol was increasingly imported and converted into bile acids. The inflammatory genes that were altered included several classic acute phase response markers, as well as genes involved in the complement, kinin, clotting, and fibrinolytic protein systems. These temporally coordinated changes in gene expression were also corroborated by biochemical measurements for FA, TG, cholesterol, and ATP. Together, these data indicate that FA are increasingly imported and oxidized in the liver to meet the enhanced energy demands arising from an inflammatory response during the first 24 h after burn injury. PMID- 15114002 TI - Carbon monoxide: to boldly go where NO has gone before. AB - The discovery that nitric oxide (NO) has powerful vasoactive properties identical to those of endothelial-derived relaxing factor spawned a vast body of research investigating the physiological actions of small gas molecules. NO, which arises endogenously through the action of nitric oxide synthase (NOS) enzymes, is a highly reactive gas that plays important roles in the regulation of vascular and immune function. Carbon monoxide (CO), a similar yet much more chemically stable gas, occurs in nature as a product of the oxidation or combustion of organic materials. CO also arises in cells and tissues as a byproduct of heme oxygenase (HO) activity, which degrades heme to biliverdin-IXalpha. Like NO, CO acts as a vasorelaxant and may regulate other vascular functions such as platelet aggregation and smooth muscle proliferation. CO has also been implicated as a neurotransmitter in the central nervous system. HO-1, the inducible form of HO, confers cytoprotection against oxidative stress in vitro and in vivo. CO, when applied at low concentration, exerts potent cytoprotective effects mimicking those of HO-1 induction, including down-regulation of inflammation and suppression of apoptosis. Many of the effects of CO depend on the activation of guanylate cyclase, which generates guanosine 3',5'-monophosphate (cGMP), and the modulation of mitogen-activated protein kinase (MAPK) signaling pathways. This review highlights new advances in the interaction of CO with cellular signaling processes. PMID- 15114003 TI - Working safely with vaccinia virus: laboratory technique and the role of vaccinia vaccination. AB - Vaccinia virus, the prototype Orthopoxvirus, is widely used in the laboratory as a model system to study various aspects of viral biology, virus-host interactions, and as a protein expression system and a vaccine vector. The ubiquitous use of vaccinia viruses in the laboratory raises certain safety concerns, because the virus can be a pathogen in individuals with immunological and dermatological abnormalities and, on occasion, can cause serious problems in normal hosts. This chapter reviews standard operating procedures when working with vaccinia virus and issues surrounding the use of prophylactic smallpox vaccination for laboratory workers. PMID- 15114004 TI - Construction and isolation of recombinant vaccinia virus using genetic markers. AB - The standard approach for the isolation of vaccinia virus recombinants involves homologous recombination between a transfected plasmid and the replicating viral DNA. In a typical infection/transfection experiment, recombinant viruses only account for a tiny proportion (10-4 to 10-3) of the progeny virus; thus, genetic markers are often included in the transfected plasmid to facilitate the selection of recombinant viruses. This chapter describes in detail two different selection procedures: one relies on plaque formation phenotype using the vaccinia virus gene F13L; the other relies on antibiotic resistance using the Escherichia coli xanthine-guanine phosphoribosyl transferase gene. PMID- 15114005 TI - Construction of recombinant vaccinia virus: cloning into the thymidine kinase locus. AB - Vaccinia virus (VV) has proven to be a very useful tool for the expression and analysis of foreign gene products. The most common method used to produce recombinant viruses involves the insertion of foreign genes into the thymidine kinase (TK) gene of the VV via homologous recombination. This is accomplished through the construction of a recombination plasmid containing the VV TK gene into the middle of which the gene of interest is inserted, appended to an efficient VV promoter element of the desired temporal class. Confluent monolayers of cells are infected with wild-type VV and transfected with the plasmid DNA to allow homologous recombination to occur. This inactivates the endogenous TK gene producing TK-negative virus that can be biochemically selected, and recombinants can be identified by a variety of screening methods. PMID- 15114006 TI - Transient and inducible expression of vaccinia/T7 recombinant viruses. AB - Recombinant DNA technology has made it possible to develop molecular cloning vectors that allow the expression of heterologous genes in a variety of animal viruses. This chapter discusses the use of vaccinia virus encoding bacteriophage T7 RNA polymerase as an expression vector system. A chosen gene is inserted into a plasmid vector designed to express genes under the control of the T7 promoter. Transient expression can then be achieved either by transfecting this plasmid into cells infected with the recombinant vaccinia virus expressing T7 RNA polymerase, vTF7-3 or by crossing this plasmid into the vaccinia virus genome and coinfecting cells with both viruses. Moreover, placement of lacO downstream of the vaccinia virus P11 late promoter regulating T7 RNA polymerase expression, and integration of lacI under vaccinia promoter control into the viral genome, vT7lacOI, yielded a recombinant virus capable of IPTG-inducible T7 promoter controlled expression of foreign genes. PMID- 15114007 TI - Construction of recombinant vaccinia viruses using leporipoxvirus-catalyzed recombination and reactivation of orthopoxvirus DNA. AB - Poxvirus DNA is not infectious because the initiation of the infective process requires proteins encapsidated along with the virus genome. However, infectious virus can be produced if purified poxvirus DNA is transfected into cells previously infected with another poxvirus. This process is termed heterologous reactivation if the infecting virus is different from the transfected virus. We describe a method in which the high-frequency recombination and replication reactions catalyzed by the Leporipoxvirus, Shope fibroma virus (SFV), can be coupled with SFV-promoted reactivation reactions to rapidly construct recombinant vaccinia viruses in high yields (25-100% recombinant progeny). The reactivated vaccinia viruses are easily purified free of the SFV helper virus by plating mixed populations of virus on cells that support only the growth of vaccinia virus. These heterologous reactivation reactions can be used to manipulate the structure of virus genomes and produce viruses that express recombinant proteins at high levels. We illustrate the method by polymerase chain reaction (PCR) cloning the gene encoding green fluorescent protein (GFP), then using double strand break repair reactions to produce a recombinant virus that expresses high levels of GFP. PMID- 15114008 TI - Construction of cDNA libraries in vaccinia virus. AB - Poxvirus expression vectors have gained widespread use for expression of foreign proteins and as delivery vehicles for vaccine antigens. We have developed a novel method using the poxvirus as a library vector for functional selection of specific cDNA. Poxviruses have several unique and useful properties as a library vector. Most importantly, because poxviruses are packaged into fully infectious particles in the cell cytoplasm, specific recombinants can be readily recovered even from a very small number of selected cells. Moreover, in contrast to libraries constructed in retrovirus or plasmid-based vectors, recombinant vaccinia virus can be efficiently recovered even from cells that have been induced to undergo apoptosis or cessation of cell growth. In the past, the major obstacle in this application to poxviruses has been the low frequency with which recombinants can be generated. The most commonly used method to construct recombinant poxvirus is homologous recombination. The frequency of recombinants derived in this manner is of the order of 0.1%, sufficient to recover a recombinant of a purified DNA clone in a transfer plasmid, but far too low to permit construction of a representative cDNA library. We have developed a method that generates nearly 100% recombinant vaccinia viruses at good titer. We have termed this method trimolecular recombination. cDNA libraries of as many as 107 or more independent viral recombinants can be constructed by trimolecular recombination. For the first time, large, diverse, and representative cDNA libraries can be screened in a vaccinia virus-based expression vector. PMID- 15114009 TI - Construction and isolation of recombinant MVA. AB - Modified vaccinia virus Ankara (MVA) is a valuable tool for the expression of recombinant genes used for such purposes as the study of protein functions or characterization of cellular and humoral immune responses. A major advantage of MVA is its clear safety record, and it can be handled under biosafety level 1 conditions. Despite its replication deficiency in human and most mammalian cells, MVA provides high-level gene expression and has proven to be immunogenic when delivering heterologous antigens in animals and humans. This chapter provides state-of-the-art protocols for generation, plaque isolation, molecular characterization, as well as amplification and purification of MVA vector viruses to obtain recombinant viruses for further evaluation. PMID- 15114010 TI - Growing poxviruses and determining virus titer. AB - Poxviruses are cell-associated viruses that can be grown in adherent- or suspension-cell cultures or chorioallantoic membranes of embryonated hen's eggs. The main principle of isolating the virus is to mechanically lyse infected cells. The virus can then be semipurified by centrifugation through a sucrose cushion. Further purification can be achieved with a sucrose gradient. This chapter describes methods for the preparation of large-scale growth and purification of the virus. PMID- 15114011 TI - Rapid preparation of vaccinia virus DNA template for analysis and cloning by PCR. AB - This chapter describes the preparation of template DNA from poxvirus-infected cells, plaques, or crude virus stocks for polymerase chain reaction (PCR) amplification. The advantages of this technique are that it is rapid, inexpensive, and, most importantly, reliable, requiring only centrifugation, detergent, and protease treatment. The template preparation is suitable for PCR amplification for screening viruses, cloning, transfection, and DNA sequencing. PMID- 15114012 TI - Orthopoxvirus diagnostics. AB - Biologic and antigenic properties are often useful for identifying and differentiating orthopoxviruses (OPV). However, polymerase chain reaction (PCR) amplification, with either restriction cleavage or sequencing of amplicons, has been gaining credibility as a more rapid, specific, sensitive, and often cost saving technique for research and diagnostic laboratories. This chapter is consolidated using prior research papers from our laboratories with three different methods that should be suitable for the preparation of orthopoxvirus DNA from various sources (e.g., clinical specimens or cell cultures) and four different methods for PCR that should be useful for investigating orthopoxvirus species and strains. PMID- 15114013 TI - An in vitro transcription system for studying vaccinia virus early genes. AB - Transcription of the vaccinia virus early genes occurs within the confines of the virion core structure. Therefore, isolated virions are a particularly rich source of proteins that function in early mRNA biosynthesis. Methods are described here for the extraction of purified vaccinia virions to yield protein mixtures with high transcriptional activity on viral early gene templates, responding specifically to both transcriptional initiation and termination signals in the DNA. PMID- 15114014 TI - An in vitro transcription system for studying vaccinia virus late genes. AB - This chapter describes a protocol that allows accurate in vitro transcription of vaccinia virus late genes. In this method, extracts are made from vaccinia virus infected cells and used as enzyme sources to produce mRNAs from plasmid templates containing late gene promoter sequences. PMID- 15114015 TI - Studying vaccinia virus RNA processing in vitro. AB - The study and use of vaccinia virus-derived RNA-modifying proteins has made a significant contribution to molecular biology. Here, the purification and assay of two such proteins, comprising the vaccinia poly(A) polymerase/cap-specific mRNA 2'-O-methyltransferase, is described. PMID- 15114016 TI - Methods for analysis of poxvirus DNA replication. AB - Cytoplasmic replication of poxviruses dictates the encoding of most, if not all, of the trans-acting factors required for faithful genome duplication. Several of these proteins have been identified through genetic and biochemical evaluation, including the catalytic DNA polymerase (E9), an essential and stoichiometric component of the processive polymerase (A20), a single-strand DNA-binding protein (I3), a type I topoisomerase (H6), the uracil DNA glycosylase (D4), a nucleic acid-independent nucleoside triphosphatase (D5), a serine/threonine protein kinase (B1), and a Holliday Junction resolvase (A22). All of these factors work in concert to faithfully duplicate the viral genome. Although a replication origin has not been defined for the poxviruses, cis-acting sequences found within the telomeric 200 bp have been implicated as necessary and sufficient for minichromosome replication. Replication occurs within cytoplasmic foci from approx 3 to 12 h postinfection. This chapter includes several methodologies to assay and quantitate replication in vivo, visualize replication foci microscopically, and test the integrity of central replication enzymes in vitro. PMID- 15114017 TI - Studying the binding and entry of the intracellular and extracellular enveloped forms of vaccinia virus. AB - This chapter describes the methods for the study of binding and entry of the two different forms of vaccinia virus (VV)-the intracellular mature virus (IMV) and extracellular enveloped virus (EEV)-using immunofluorescent staining and confocal microscopy. After binding to or penetration of the cells, IMV, EEV, and virus cores are distinguished by different antibodies. Bound virus or penetrated cores are visualized and recorded by confocal microscopy and can be accurately counted. Although specific antibodies to IMV, EEV, and virus cores are required, this method is highly quantitative and also allows the recognition of virus aggregates, which would not be possible using other techniques, such as flow cytometry and radiolabeling of virus particles. Furthermore, this method bypasses the need for EEV purification that may damage the EEV membrane and release an IMV particle. PMID- 15114018 TI - Pox, dyes, and videotape: making movies of GFP-labeled vaccinia virus. AB - The large size of poxvirus virions (approx 250 x 350 nm) makes them ideal candidates for microscopic studies. Recombinant vaccinia viruses that express a viral envelope-specific, green fluorescent protein (GFP) chimera produce enveloped virions that fluoresce green. This fluorescent labeling allows the live, real-time study of viral egress using a variety of microscopic techniques. The methods presented here describe how to image the movement of intracellular enveloped virions that are labeled with green fluorescent protein using time lapse laser scanning confocal microscopy. Details are also provided for analyzing the images obtained and converting them into QuickTime movies suitable for presentation. PMID- 15114019 TI - Interaction analysis of viral cytokine-binding proteins using surface plasmon resonance. AB - Surface plasmon resonance (SPR) biosensors have become an increasingly popular technology for characterizing the protein-protein interactions of virus-host interactions. Various studies have exploited the versatility of SPR to probe the interaction between virus and host components, including constituents of virus particles and host cellular receptors, as well as interactions between viral proteins and host immune molecules. This chapter describes basic procedures for employing SPR to study the interaction between poxvirus proteins and host immune signaling proteins. We also identify how this methodology may be adapted toward other applications relevant in the study of poxvirus-host interactions. PMID- 15114020 TI - Monitoring of human immunological responses to vaccinia virus. AB - For the last 30 yr, interest in vaccinia virus immune monitoring has focused on the use of the vaccinia virus as a recombinant vaccine vector and the potential detrimental effect of antivector immunity on subsequent vaccination with a recombinant vaccinia virus. However, interest in this area has intensified after the publication of reports suggesting that smallpox may be a major pathogen selected for bioterrorist activities. Owing to the unacceptably high incidence of complications induced by previous effective smallpox vaccine strains, alternative safer strains (e.g., modified vaccinia Ankara [MVA]) are being assessed for their antigenicity in clinical trials. The exact immune effector mechanism responsible for vaccine-induced protection to smallpox infection has not been fully elucidated, although it is believed that neutralizing antibody plays a major role. This chapter describes a simple enzyme-linked immunosorbent assay (ELISA) to quantify vaccinia virus antibody titer. Additionally, to define serum neutralizing activity, both a classical plaque reduction assay and a high throughput 96-well plate method based on reduction of recombinant vaccinia virus expressed beta-galactosidase is described. Furthermore, details are given for a T cell proliferation assay, primarily for monitoring T-helper CD4 activity and an enzyme-linked immunospot (ELISPOT) assay for CD8 analysis. The use of reliable immunological assays is vital in assessing the potential efficacy of new vaccines to protect against smallpox infection. PMID- 15114021 TI - Vaccinia virus as a tool for immunologic studies. AB - Studies that involve antigen processing and presentation often require de novo biosynthesis of the antigen both in vitro and in vivo. Additionally, biosynthesis of the antigen or engineered variants within the antigen-presenting cells is usually simpler than providing purified recombinant proteins from bacteria, yeast, or insect cells. For these purposes, recombinant vaccinia virus-based expression has several advantages over other expression systems employed in the field. Insertion of large pieces of recombinant DNA into the vaccinia virus genome, easy recombination and selection of vaccinia viruses, and the ability of these viruses to infect a variety of cells are some key aspects that have made this system popular. Although their efficacy is proven in studies of major histocompatibility complex (MHC) class I-restricted antigen processing and presentation, it is challenging to use them in MHC class II-restricted antigen processing and presentation owing to many reasons specified in this chapter. This chapter aims to describe the commonly used procedures in this field that employ vaccinia virus systems, particularly troubleshooting common problems encountered during experiments. PMID- 15114022 TI - Mouse models for studying orthopoxvirus respiratory infections. AB - Concern regarding the use of variola and monkeypox viruses as bioterrorist agents has led to an increased study of orthopoxviruses to understand the molecular and cellular basis of pathogenesis and develop safe and effective antivirals and vaccines against smallpox. Crucial to these efforts is the availability of animal models, which are inexpensive, genetically homogeneous, and recapitulate the human disease. The popular small-animal orthopoxvirus models employ the inbred mouse as the host, the respiratory tract as the site of virus inoculation, and orthopoxviruses-vaccinia, cowpox, and ectromelia viruses-as surrogates for variola virus. Ectromelia virus is likely the best surrogate for variola virus in a mouse model, as it is infectious at very low doses of virus, and the mousepox disease is associated with high mortality in the susceptible A, BALB/c, and DBA/2 stains of mice, but causes an unapparent infection in the C57BL/6 mouse strain. This chapter describes an ectromelia virus respiratory infection model in the mouse. PMID- 15114023 TI - Viral glycoprotein-mediated cell fusion assays using vaccinia virus vectors. AB - The vaccinia virus-based expression of viral envelope glycoprotein genes-derived from enveloped viruses that infect their respective host cells through a pH independent mechanism of membrane fusion-has been a powerful tool in helping to characterize these important attachment and fusion proteins. The cellular expression of these viral envelope glycoproteins has allowed for the measurement of membrane fusion events using cell-cell fusion or syncytia formation. This method has been enhanced by the addition of a reporter-gene system to the vaccinia virus-based cell-cell fusion assay. This improvement has provided a high throughput and quantitative aspect to this assay, which can serve as a surrogate for virus entry and is therefore ideally suited in the characterization of numerous enveloped viruses, including biological safety level-4 (BSL-4) agents. This chapter will detail the methods of the vaccinia virus-based reporter-gene fusion assay and how it may be used to characterize the fusion mediated by the BSL-4-classified Hendra and Nipah viruses. PMID- 15114024 TI - Use of dual recombinant vaccinia virus vectors to assay viral glycoprotein mediated fusion with transfection-resistant primary cell targets. AB - Fusion mediated by the human immunodeficiency virus type-1 (HIV-1) envelope (Env) glycoprotein and the cellular CD4/chemokine receptor complex is the first step in entry and is often analyzed in cell-cell fusion assays that require Env expression by recombinant vaccinia viruses and/or target cell transfection. Primary lymphocytes and macrophages are the principal targets for HIV-1 in vivo, but are poor substrates for transfection, and constructing recombinant vaccinia viruses expressing every novel or mutant env gene is laborious. This chapter describes a fusion assay using two recombinant vaccinia viruses that express distinct RNA polymerases suitable for transfection-resistant targets, such as primary human lymphocytes and macrophages. It also uses env genes contained in plasmid vectors, eliminating the need to construct recombinant vaccinia viruses to analyze each construct. Effector 293T cells are cotransfected with SP6-driven reporter gene and T7-driven env plasmids, then infected with recombinant vaccinia virus expressing T7 polymerase. Primary lymphocyte or macrophage targets are infected with recombinant vaccinia virus expressing SP6 polymerase. Fusion mediated by effector cell Env and endogenous CD4/coreceptors in target lymphocytes or macrophages enables SP6 polymerase-mediated reporter gene transactivation. This approach provides an efficient tool to study fusion mediated by multiple-cloned primary isolate or mutant HIV-1 env genes with the primary target cell types relevant to infection in vivo. PMID- 15114025 TI - Poxvirus bioinformatics. AB - Biochemical and functional analysis of poxvirus genomes, genes, and proteins has entered a new era with the recent sequencing of more than 30 poxvirus genomes. The management and analysis of this volume of sequence data in an efficient and effective manner requires specialized computer software. This chapter describes a number of bioinformatics techniques useful for analyzing poxvirus genomes. Some of the software discussed here have been developed by members of the Poxvirus Bioinformatics Resource Center (PBRC; funded by National Institutes of Health [NIH]) specifically for use with poxvirus genomes. These programs or, more accurately, suites of programs have many functions dedicated to poxvirus genome characterization. Significantly, this software has been designed with ease of use at a single location as the major goal. PMID- 15114026 TI - Preparation and use of molluscum contagiosum virus from human tissue biopsy specimens. AB - Molluscum contagiosum virus (MCV) lesions are limited to the human epidermis and can persist for months, showing only weak signs of inflammation. Because a culture system has not yet been created to replicate MCV, this chapter describes how the virus can be isolated from patient specimens. From this material, we describe how MCV is purified and used for infection studies, electron microscopy, viral DNA extraction, and analyses of early mRNA synthesized by in vitro transcription of permeabilized virions. The complete MCV-1 genome has been sequenced, and a redundant MCV genome fragment library of MCV type 1 (available from American Type Culture Collection [ATCC]) is useful for the cloning and study of individual MCV genes. PMID- 15114028 TI - Radial endoscopic ultrasound and spiral computed tomography in the diagnosis and staging of periampullary tumors. AB - BACKGROUND AND STUDY AIMS: The early detection and accurate staging of pancreatic and ampullary cancer is of utmost importance for the achievement of surgical radical treatment. The aim of this study was to assess prospectively the role of endoscopic ultrasonography (EUS) in detection and staging of pancreatic and ampullary cancer, comparing its results to those obtained with spiral computed tomography (SCT). PATIENTS AND METHODS: Sixty-one patients with suspected pancreatic and ampullary tumors were included, 46 (75, 4%) of whom presented with obstructive jaundice. Patients underwent EUS and SCT within a 7-day period. Examiners were unaware of the previous imaging results, except conventional echography. Image interpretation was compared to surgical and histopathological findings. RESULTS: Fifty-six (91, 8%) patients were surgically explored. Clinical follow-up and/or tissue diagnosis determined the correct diagnosis in the remaining five patients. Pancreatic cancer and ampullary cancer were observed in 29 (47, 6%) and 10 (16, 4%) patients, respectively. Chronic pancreatitis and choledocholithiasis were the most common diagnosis in patients with non neoplastic disease. EUS was more effective than SCT for the definition of the final diagnosis in patients with obstructive jaundice (87.0 vs. 67.4%, p = 0.04). Both exams were equally effective for detecting pancreatic cancer but EUS predicted more accurately the involvement of portal-mesenteric axis by the tumor (87.0 vs. 67.4%, p = 0.04). EUS was particularly useful in the diagnosis of cancer of papilla of Vater. CONCLUSION: In patients with pancreatic adenocarcinoma without unequivocal signs of distant metastatic disease, EUS is more accurate than SCT to predict venous involvement by the tumor. EUS is superior to SCT to detect ampullary adenocarcinoma. Both methods are equally ineffective to detect nodal involvement in pancreatic and ampullary cancer. PMID- 15114029 TI - Individualized risk management in diabetics: how to implement best practice guidelines--design and concept of the IRIDIEM studies. AB - The prevalence of type 2 diabetes mellitus is rising rapidly in all developed countries, particularly in the growing population of persons >50 years of age. As a dangerous consequence, this is accompanied by a proportionate increase in the incidence of chronic renal disease. Evidence-based medicine has shown that tight blood glucose control can delay the onset and retard the progression of diabetic complications, and while it is a challenge to closely manage the complexity of diabetes, it is more difficult to effectively treat the multiple associated comorbidities that develop. Best practice guidelines support early intervention and aggressive treatment of hypertension, hyperglycaemia, proteinuria, hypercholesterolemia, and anaemia. To date, guideline-based management has been proven to be difficult. This article describes the concept of the IRIDIEM studies. The objective of these studies is to endorse and facilitate the use of current best practice guidelines for the management of frequent comorbid diseases and established risk factors in the treatment of type 2 diabetes associated with chronic kidney disease. Additionally, IRIDIEM will assess the impact of this improved disease management model on the progression of chronic kidney disease that can result from electronically prompting clinicians with evidence-based treatment advice. PMID- 15114030 TI - Chlamydia pneumoniae and atherosclerosis: no way-out or long way? What about renal failure patients? AB - Recently, Chlamydia pneumoniae is the microorganism frequently implicated in the infection-based inflammatory atherogenous hypothesis. Although in vitro experimental data and initial sero-epidemiologic, pathology-based studies and antibiotic trials supported this interesting hypothesis, later data are conflicting. Some confounding factors are the causes of uncertainty; lacking of standard methods for C. pneumoniae detection, co-existence of other atherosclerotic risk factors and anti-inflammatory effects of antibiotics used in clinical trials seem to be the principal ones. Standardization of methodology used, antibiotic trials with a different orientation-design and a vaccine preparation that eventually will be tested in clinical trials with a long follow up, should provide a definite answer regarding the probability C. pneumoniae to be a main, a secondary or an irrelevant factor to atherosclerosis. Studies linking C. pneumoniae to inflammation and accelerated atherosclerosis in renal failure patients are accumulated but limitations are similar to the above mentioned. PMID- 15114031 TI - Effect of IgG therapy on lymphocyte subpopulations in the peripheral blood of Kuwaiti women experiencing recurrent pregnancy loss. AB - Intravenously administered polyspecific IgG is being increasingly used as an immunomodulating therapy with controversial beneficial outcome. The aim of this study was to evaluate the effects of IgG infusion on peripheral T-cell subpopulations in women with recurrent pregnancy loss (RPL). Fifteen women with a history of three previous RPL between 6 and 22 weeks of gestation and positivity for the antiphospholipid antibody syndrome (APS) were randomized to one of two treatment groups: (a) an intravenous immunoglobulin therapy group (RPL-IVIg; 7 patients), 500 mg IVIg/kg/month and (b) a placebo-treated group given multivitamins (8 patients). Control groups comprised either normal pregnant women without APS (10 patients) or non-pregnant women. The T-cell markers were characterized using a monoclonal antibody panel including CD3, CD4, CD8, CD25, CD29, CD38, CD45RA, CD45RO, CD54 and HLA-DR. Analysis was performed with a two color fluorescent-activated flow cytometer. In the first trimester, the percentage of CD4+CD25+, CD4+CD45RO+, CD8+HLA-DR+, and CD8+CD38+ populations were reduced in the multivitamin group compared to normal pregnant women (p < 0.05) while in the RPL-IVIg group only CD4+CD25+ cells were reduced (p < 0.05). By the second trimester, CD3+CD16+CD56+ was significantly higher in multivitamin- than in IVIg-treated women (p < 0.05). The percentage of CD4+HLA-DR+ was significantly higher in the two RPL groups compared to normal pregnant women (p < 0.05). IVIg therapy in women with RPL was associated with a significant reduction in CD3+CD16+CD56+ and CD4+CD25+. This may contribute to the suppression of immune mediated processes contributing to premature abortion. PMID- 15114032 TI - 'The multiple-sit-to-stand' field test for older adults: what does it measure? AB - BACKGROUND: The need to predict decline in functional status in a large number of older adults has brought researchers and clinicians to develop easy-to-administer field tests. One of them is the 'multiple-sit-to-stand' (MSTS), which claims to measure leg strength. OBJECTIVE: To assess the extent to which the MSTS is a leg strength, leg endurance or general endurance test. METHODS: 49 independently functioning women (72.2 +/- 6.4 years) were assessed on the MSTS, on a submaximal stress test, and on strength and endurance of knee extensors measured by isokinetics. The knee extensors were selected, more than in other muscle groups responsible for movement of the lower extremity, as the largest range of motion, and the largest torques required while performing activities of daily living (ADL) is found in this group. RESULTS: The correlation of the MSTS with the stress test was moderate and significant but very low and nonsignificant with the isokinetic measurements. CONCLUSIONS: The MTST is not able to predict strength of knee extensors, the leading group of leg muscles in ADL. If at all, it predicts general endurance rather than any measure--muscle strength or muscle endurance- of lower extremities. Longitudinal studies assessing the potential of the MSTS to predict deterioration in ADL in older adults are recommended, as well as studies assessing other factors related to both MSTS and ADL, such as muscle-nerve coordination affecting multiple joint activities. PMID- 15114033 TI - Age-dependent expression of advanced glycation end product receptor genes in the human heart. AB - BACKGROUND: Advanced glycation end products (AGEs) are formed by the reaction of sugars and NH2 groups of lysine and arginine residues and have been shown to accumulate in tissues, including the heart, with normal ageing. The interaction of AGEs with their receptors is known to cause changes in cell function, leading, for example, to the production of pro-inflammatory cytokines and free radicals. OBJECTIVE: This study investigated the gene expression of the five known AGE receptors: AGE-R1, AGE-R2, AGE-R3, the scavenger receptor II, and the receptor for AGEs (RAGE) in human heart tissue. METHODS: Tissue samples were taken from the right cardiac auricles from three patient groups: children (2.4 +/- 1.1 years), adults (45.3 +/- 0.8 years) and elderly subjects (76.4 +/- 0.4 years). Analysis of gene expression of the five AGE receptors was performed using the reverse transcription-polymerase chain reaction (RT-PCR) and 18S mRNA levels as loading controls. RESULTS: Our results show an age-dependent upregulation of the genes for AGE-R3 and the scavenger receptor II, but a downregulation for RAGE and no significant differences for AGE-R1 and AGE-R2. CONCLUSION: This study supports a pathophysiological function for AGE receptors such as AGE-R3 and RAGE in the ageing heart. PMID- 15114034 TI - Physical function and health status among seniors with and without a fear of falling. AB - BACKGROUND: Fear of falling (FF) is pervasive among older people and is an independent risk factor for decreased mobility and loss of quality of life. Although it is a serious health concern, little is known about the severity of the physical and health-related deficits in high-functioning fearful seniors. OBJECTIVE: This study examined the physical correlates of FF in community dwelling seniors in relation to their non-fearful counterparts and also explored the relevance of recent fall history. SUBJECTS: Twenty-five volunteers who stated that they had curtailed their activities due to a FF and who reported themselves to be healthy participated. Non-fearful, age- and gender-matched subjects were also recruited and served as a comparison group. METHODS: This cross-sectional study had participants come to the laboratory on one occasion for about 1.5 h. Limits of balance stability, walking speed, and lower limb muscle strength were measured in random order. In addition, the SF-36, the Activities-specific Balance Confidence (ABC) scale and the Human Activity Profile questionnaires were completed to provide measures of heath status, FF and activity levels, respectively. RESULTS: Subjects with a FF had lower ABC scores (69.8 +/- 10.2) than controls (90.2 +/- 7.9; p < 0.001), and although they reported activity curtailment, the groups presented similar activity profiles (p > 0.05). Balance ability was not compromised in seniors with a FF, although their walking speed was slower than that of control subjects (p < 0.004). Lower limb weakness was significant in the FF group, which also reported low physical health. Perceived physical health, activity and hip flexor torque in combination accounted for almost 62% of the variance in ABC scores. Secondary analysis of the data from the FF group revealed a tendency for those who had fallen in the past year to restrict their limits of stability compared to those who had not fallen; though generally the effect sizes were small to moderate. CONCLUSIONS: The marked deficits in strength and health status found among seniors living independently in the community, who are in good health, but report being fearful of falling underscores the seriousness of FF as a potential health risk factor in the well elderly. PMID- 15114035 TI - Effect of aging on visual shape distortion. AB - BACKGROUND: Individuals experience a visual illusion created by shape interaction: when two shapes are presented successively and briefly, the form of the second (test) shape appears distorted due to the form of the first (prime) shape; this shape interaction is called the shape distortion effect. While age related deterioration in performance is found in various aspects of visual perception, the effect of aging on the shape distortion effect has not been evaluated. OBJECTIVE: The purpose of this study is to determine the effect of aging on the shape distortion effect. METHODS: We measured the perception of briefly presented elementary shapes and of the shape distortion effect in 29 healthy volunteers, with ages ranging from 18 to 83 years. For each shape interaction trial, a prime rectangle was presented (vertical or horizontal) for 45 ms, followed by an interstimulus interval for 135 ms, a test circle for 60 ms and finally a random dot mask for 300 ms. The test circle was presented in each quadrant of the visual field. The prime rectangle was flashed either at the same position as the test circle (0 degrees offset, intrahemifield) or 11 degrees apart, displaced horizontally in the opposite hemifield (11 degrees offset, interhemifield). In the elementary-shape trials, there was no prime, and the test shape was a circle or an ellipse. Using the method of adjustment, the percent elongation [(longer diameter - shorter diameter)/(shorter diameter) x100] of the reproduced ellipse was computed. RESULTS: The mean percent elongation in response to the elementary shapes did not vary with increasing age. The shape distortion effect decreased significantly with increasing age during both intra- and interhemifield conditions. The mean shape distortion effect was larger for the intrahemifield condition than for the interhemifield one. CONCLUSION: The shape distortion effect decreases with advancing age while the perception of elementary shapes does not. These results indicate a severer age-related dysfunction of the cerebral processing of shape interaction than that of elementary-shape perception. PMID- 15114036 TI - Memory impairment means less pain for mice. AB - BACKGROUND: Clinical observations have reported that individuals with memory deterioration, like in Alzheimer's disease, display a lesser pain sensibility than patients with no cognitive impairment. OBJECTIVE: To clarify the link between pain and loss of memory, we studied how memory-impaired mice behave when submitted to hotplate nociceptive tests. METHODS: For 5 days (D1-D5), male CD1 mice were injected daily intraperitonealy with saline or scopolamine (s, an anticholinergic drug, 0.2 mg/kg) or ketamine (k, an N-methyl-D-aspartate receptor antagonist (NMDAr), 2.5 mg/kg), at doses leading to memory impairment with no analgesic effect. From D6 to D9, all received saline only. They were placed on the hotplate and removed at the first sign of discomfort, response time being recorded. RESULTS: From D1 to D5, reaction time decreased significantly in controls only and did not change in mice with scopolamine or ketamine. From D6 to D9, response times decreased (p < 0.05 (s) and p < 0.0001 (k)) to reach the steady state of control animals. At D5, response time was significantly prolonged for scopolamine (p < 0.01) and ketamine (p < 0.05), compared to controls. CONCLUSION: These results show that pain sensibility needs the integrity of the central cholinergic and of the NMDA systems, and that mice with memory impairment display a lesser pain sensibility than normal mice. Further research on the complex interactions of receptors and neurotransmitters involved in pain and cognition could assist in gaining a better understanding of pain and analgesia in patients with memory impairment and in demented individuals. PMID- 15114037 TI - Does practice modify the relationship between postural control and the execution of a secondary task in young and older individuals? AB - BACKGROUND: Numerous daily activities require performing more than one task simultaneously, such as standing while engaging in a conversation. Recent studies have shown that postural control may be degraded when individuals are asked to perform a secondary task and that this effect seems to be more pronounced in older adults. Since various types of secondary tasks have been used in postural control studies, the novelty of the tasks may partly explain why dual-task interference occurs. It is known that novel tasks require greater attentional resources and thus may interfere to a greater extent with the performance of another task. Therefore, by practicing this dual-task situation, interference could perhaps be diminished. Since the dual-tasking efficiency is reduced with aging, practice could be very beneficial to older adults. OBJECTIVES: The main goal of this study was to examine whether practice could modify the changes seen in postural sway when individuals are asked to perform a secondary task while maintaining upright stance and whether older individuals could benefit to a greater extent from practice than would young individuals. The second goal was to examine the dual-task performance in young versus older adults and to determine whether older individuals benefit equally or to a greater extent from practice as compared with young individuals. METHODS: Young and older individuals were asked to stand on a force platform while performing a secondary task or no task. The secondary task condition was repeated six times to examine the effects of practice. RESULTS: Practice did not modify the performance of postural sway, but did lead to an increase in speed of execution of the secondary task for both groups equally. In young participants, the amplitude of sway was decreased, and the frequency of sway was increased, indicating an increased stiffness when performing the cognitive task. Older participants showed increased amplitude of sway and increased frequency of sway in the mediolateral direction only. CONCLUSIONS: Since the dual-task condition was only repeated six times, it could be hypothesized that the effect of practice would be greater, if more trials were added or if more practice sessions were included. More research is needed to verify this hypothesis. PMID- 15114038 TI - Assessing life expectancies of institutionalized older persons in Hong Kong. AB - BACKGROUND: In Hong Kong, it has been projected that, by the year 2010, there will be 0.86 million persons aged 65 or older. The demand for residential care places continues to rise. Information on the survival of older persons will allow better management of care in institutional settings. OBJECTIVE: The Weibull model was developed to predict the 18-month survival of a group of Chinese nursing home residents. METHODS: In this prospective cohort study, 590 older residents from four nursing homes with different levels of care were enrolled. The main outcome measure was survival at 18 months. Information on subject demographics, clinical data and disability levels as measured by the Patient Assessment Instrument (PAI) was collected at baseline. Life expectancies were derived from accelerated failure time models. RESULTS: Independent predictors of decreased survival derived from the Weibull model included increased age, male gender, diagnosis of cancer, presence of malnutrition and functional dependence. Significant interaction was observed between male gender and PAI score. The goodness of fit of the model was satisfactory. CONCLUSIONS: From this study, the model for predicting life expectancy in nursing home residents can assist policy makers in planning long-term care. It can also guide clinicians to make more appropriate management decisions for their older patients. PMID- 15114039 TI - One-year incidence of hyperosmolar states and prognosis in a geriatric acute care unit. AB - BACKGROUND: Hyperosmolar syndromes are associated with high mortality rates, yet little is known about their incidence and their prognosis. OBJECTIVE: To determine the 1-year incidence of hyperosmolar states and the prognostic factors for in-hospital and 1-year mortality. METHOD: A 6-month prospective cohort study was conducted in a 40-bed acute care geriatric unit and included all patients who developed plasma osmolarity of 320 mosm/l or greater. Age, sex and known cognitive impairment as possible risk factors of hyperosmolarity were assessed. In-hospital and 1-year mortality were calculated and risk factors for death among baseline patient characteristics were sought. RESULTS: 48 (11) of the 436 inpatients in the study were identified as hyperosmolar. Diabetic hyperosmolarity was found in 8 patients. Cognitive impairment was a risk factor for hyperosmolarity (relative risk 2.39, 95% confidence interval 2.18-3.33, p < 0.001), but not age or sex. Infections were accompanied by hyperosmolarity in 30 (62.5). Thirty-five patients (72.9) were bed- or chair-ridden. In-hospital mortality was higher in hyperosmolar patients (35.4) than in the others (16.7%, p = 0.003). Causes of death were infection in 5 (29.4), terminal cachexia in 5, thrombosis in 3, gastric bleeding in 1, renal failure in 2 and heart failure in 1. Functional dependency for mobility was a risk factor for in-hospital mortality but not the degree of hyperosmolarity. One-year mortality was 68.7%. Functional dependency and pressure ulcers were independent predictors of 1-year mortality (p = 0.005 and p = 0.044, respectively). CONCLUSION: Hyperosmolar states occurred in cognitively impaired and dependent patients and resulted in high mortality rates at short and at mid-term. Mortality was related to functional dependency rather than to hyperosmolarity. PMID- 15114040 TI - Decline of vertical gaze and convergence with aging. AB - BACKGROUND: Disturbance of vertical eye movement and ocular convergence is often observed in elderly people, but little is known about its frequency. OBJECTIVE: The purpose of this study was to investigate age-associated changes in vertical eye movement and convergence in healthy elderly people, using a digital video camera system. METHODS: We analyzed vertical eye movements and convergence in 113 neurologically normal elderly subjects (mean age 70 years) in comparison with 20 healthy young controls (mean age 32 years). The range of vertical eye movement was analyzed quantitatively and convergence was analyzed qualitatively. RESULTS: In the elderly subjects, the angle of vertical gaze decreased with advancing age and it was significantly smaller than that of the younger subjects. The mean angle of upward gaze was significantly smaller than that of downward gaze for both young and elderly subjects. Upward gaze impairment became apparent in subjects in their 70s, and downward gaze impairment in subjects in their 60s. Disturbance in convergence also increased with advancing age, and was found in 40.7% of the elderly subjects. CONCLUSION: These findings indicate that the mechanisms of age-related change are different for upward and downward vertical gaze. Digital video camera monitoring was useful for assessing and monitoring eye movements. PMID- 15114041 TI - Heart Rate and Blood Pressure Changes during Gastroscopy in Healthy Older Subjects. AB - BACKGROUND: Previous studies uncontrolled for co-morbidity have shown increased heart rate and decreased oxygenation during gastroscopy in older people. One previous study of selected younger patients suggested that changes in blood pressure were more pronounced in non-premedicated patients. AIMS: To prospectively examine blood pressure and heart rate responses during gastroscopy in healthy older people uninfluenced by medication or co-morbidity. METHODS: Blood pressure and heart rate were monitored continuously in consecutive unselected patients attending for gastroscopy carried out by one endoscopist. RESULTS: Thirty-seven patients were included with a mean age of 51.5 years (95% CI: 17-89); 10 patients were over the age of 65. During gastroscopy there were large increases in blood pressure and heart rate, with maximum changes occurring predominantly at intubation in the younger age group. Typically, increases in blood pressure in the older group occurred later in the procedure and were more sustained. Mean (95%CI) maximum changes in systolic blood pressure were more profound in those sedated [73 (55-91) mm Hg] compared to those having throat spray [47 (34-60) mm Hg; p < 0.05]. Maximum changes in heart rate during gastroscopy became significantly smaller with advancing age in those having throat spray (p = 0.03) but not in those receiving sedation. CONCLUSIONS: Gastroscopy stimulates blood pressure and heart rate changes that are greatest in those sedated. Further studies of the effect of gastroscopy upon blood pressure are required, particularly in older patients, the group regarded as at increased risk from complications. PMID- 15114042 TI - Role of mitochondrial deterioration in physiological and pathological brain aging. AB - BACKGROUND: Mitochondria are widely reported to occupy a unique role in modulating cell viability, senescence and death. This is consistently supported by the multiple functions of these organelles. In addition to providing the energy for the myriad of cellular performances, mitochondria are involved in regulating thermogenesis, calcium buffering, integration of pro- and anti apoptotic signals. OBJECTIVE: To stress the significant importance of subtle, continuous and permanent mitochondrial alterations as key events in physiological aging and as unfavourable determinants of age-related neurodegenerative diseases. RESULTS: Any dysfunction of these organelles may constitute a serious threat for cellular health status and survival, particularly of post-mitotic nerve and muscle cells. Mitochondrial deterioration may affect discrete features of the organelles (such as their structural dynamics, genetics and physiology) and lead to a progressive functional impairment. CONCLUSIONS: A variety of mitochondrial tasks, while hampering the possibility to recover the organelles' dysfunctions, offer different and reliable opportunities for therapeutic interventions. PMID- 15114043 TI - Acupuncture and moxibustion in the treatment of active Crohn's disease: a randomized controlled study. AB - BACKGROUND: Acupuncture has traditionally been used in the treatment of inflammatory bowel disease in China and is increasingly being applied in Western countries. The purpose of this study was to investigate the efficacy of acupuncture in the treatment of active Crohn's disease (CD). METHODS: A prospective, randomized, controlled, single-blind clinical trial was carried out to analyze the change in the CD activity index (CDAI) after treatment as a main outcome measure, and the changes in quality of life and general well-being, serum markers of inflammation (alpha(1)-acid glycoprotein, C-reactive protein) as secondary outcome measures. 51 patients with mild to moderately active CD were treated in a single center for complementary medicine by three trained acupuncturists and randomly assigned to receive either traditional acupuncture (TCM group, n = 27) or control treatment at non-acupuncture points (control group, n = 24). Patients were treated in 10 sessions over a period of 4 weeks and followed up for 12 weeks. RESULTS: In the TCM group the CDAI decreased from 250 +/- 51 to 163 +/- 56 points as compared with a mean decrease from 220 +/- 42 to 181 +/- 46 points in the control group (TCM vs. control group: p = 0.003). In both groups these changes were associated with improvements in general well-being and quality of life. With regard to general well-being, traditional acupuncture was superior to control treatment (p = 0.045). alpha(1)-acid glycoprotein concentration fell significantly only in the TCM group (p = 0.046). CONCLUSIONS: Apart from a marked placebo effect, traditional acupuncture offers an additional therapeutic benefit in patients with mild to moderately active CD. PMID- 15114044 TI - Contribution of glutamine synthetase to ammonia-induced apoptosis in gastric mucosal cells. AB - BACKGROUND/AIMS: Glutamine synthetase is a key enzyme necessary for ammonia detoxification in the brain, but excessive activation of this enzyme can be cytotoxic to neural cells as a consequence of excessive consumption of ATP and glutamate. The stomach also expresses high levels of glutamine synthetase and this study aimed to investigate a possible pathophysiological role of glutamine synthetase in ammonia-induced gastric mucosal injury. METHODS: Normal rat gastric mucosal epithelial (RGM-1) cells were treated with ammonia, and a specific glutamine synthetase inhibitor (methionine sulfoximine) was used to assess the action of glutamine synthetase. RESULTS: Treatment with ammonia induced apoptotic cell death. Increased expression of p21 and Bax, decreased expression of Bcl-2, cytochrome C release from the mitochondria into the cytosol and subsequent activation of caspase-9 and -3 were identified in the cells treated with ammonia, although there was no apparent change in p53 expression. On the other hand, pretreatment with various concentrations of methionine sulfoximine reduced the glutamine synthetase activity in ammonia-treated RGM-1 cells, and prevented the induction of apoptosis and the reduction in intracellular ATP levels in a dose dependent manner. CONCLUSIONS: Our results suggested that the energy exhaustion which resulted from an overload of ammonia to glutamine synthetase may have initiated the apoptotic signaling in gastric mucosal cells. PMID- 15114045 TI - Effect of intraduodenal administration of ursodeoxycholic acid on interdigestive interaction between gallbladder motility, pancreatic secretion and endocrine activity. AB - BACKGROUND: Gastroduodenal motorfunction, gallbladder motility, and pancreatic secretion are closely related during the interdigestive state. The extent to which application of ursodeoxycholic acid (UDC) influences this process is only partly understood. AIM: As UDC is widely used for the therapy of gallbladder stones and of cholestatic liver disease, we wanted to define the immediate effect of UDC on interdigestive gallbladder and antroduodenal motility, biliary pancreatic secretion and hormone release in man. METHODS: Interdigestive gastrointestinal function in 10 healthy males (26-35 years) was studied twice after 12-hour fasting on 2 different days. Antroduodenal motility was continuously recorded manometrically over a complete interdigestive migrating motor complex (MMC) cycle. Gallbladder volume was evaluated sonographically in 5- to 7-min intervals during the MMC cycle. Pancreatic and biliary secretion was determined by a standard duodenal perfusion technique measuring chymotrypsin, amylase, lipase and bile salts in duodenal aspirates every 15 min. Plasma levels of pancreatic polypeptide (PP) and motilin were determined by radioimmunoassay in 15-min intervals. On 2 separate days, 7-10 days apart, each subject received intraduodenally either 10 mg/kg UDC (pH 8) or placebo 30 min after the first recorded duodenal MMC cycle phase III. RESULTS: With placebo, the fasting gallbladder volume decreased slightly from phase I (32 +/- 8 ml) to the end of phase II (24 +/- 13 ml), but increased significantly from 31 +/- 14 ml (phase I) to 46 +/- 11 ml (phase III) after intraduodenal UDC application (p < 0.01). Pancreatic secretion was significantly reduced after UDC application at the end of phase II (secretion of chymotrypsin 10 +/- 3 U/min vs. 5 +/- 2 U/min, p < 0.01). Serum levels of PP were also reduced by UDC during the entire MMC cycle. This reached statistical significance at the end of phase II (84 +/- 8 pg/ml vs. 57 +/- 14 pg/ml; p < 0.05) and during phase III (86 +/- 19 pg/ml vs. 64 +/- 22 pg/ml; p < 0.05), while motilin slightly increased during the MMC cycle after UDC application. UDC instillation did not affect antroduodenal motility. CONCLUSION: UDC exerts significant inhibitory effects on interdigestive gallbladder contractility, pancreatic secretion, and PP release. Whether these inhibitory effects are mediated by cholinergic pathways or other mechanisms requires further investigation. PMID- 15114046 TI - Takayasu's arteritis accompanied with pericarditis: a case report. PMID- 15114047 TI - High frequency stimulation of the subthalamic nucleus eliminates pathological thalamic rhythmicity in a computational model. AB - Deep brain stimulation (DBS) of the subthalamic nucleus (STN) or the internal segment of the globus pallidus (GPi) has recently been recognized as an important form of intervention for alleviating motor symptoms associated with Parkinson's disease, but the mechanism underlying its effectiveness remains unknown. Using a computational model, this paper considers the hypothesis that DBS works by replacing pathologically rhythmic basal ganglia output with tonic, high frequency firing. In our simulations of parkinsonian conditions, rhythmic inhibition from GPi to the thalamus compromises the ability of thalamocortical relay (TC) cells to respond to depolarizing inputs, such as sensorimotor signals. High frequency stimulation of STN regularizes GPi firing, and this restores TC responsiveness, despite the increased frequency and amplitude of GPi inhibition to thalamus that result. We provide a mathematical phase plane analysis of the mechanisms that determine TC relay capabilities in normal, parkinsonian, and DBS states in a reduced model. This analysis highlights the differences in deinactivation of the low-threshold calcium T -current that we observe in TC cells in these different conditions. Alternative scenarios involving convergence of thalamic signals in the cortex are also discussed, and predictions associated with these results, including the occurrence of rhythmic rebound bursts in certain TC cells in parkinsonian states and their drastic reduction by DBS, are stated. These results demonstrate how DBS could work by increasing firing rates of target cells, rather than shutting them down. PMID- 15114048 TI - Decoding input signals in time domain--a model approach. AB - From an observation of efferent interspike intervals of a neuron, we consider how to decode the input temporal information. It is found that the integrate-and-fire model is blind in the temporal domain due to the fact that its efferent firing rate is independent of the input temporal frequency. The conclusion is then confirmed for the integrate-and-fire model with correlated inputs, with reversal potentials, with a nonlinear leakage and with a subthreshold oscillation. For the Hodgkin-Huxley model, however, in terms of efferent firing rates alone, it is possible to read out the input temporal information. PMID- 15114049 TI - Spatial localization of synapses required for supralinear summation of action potentials and EPSPs. AB - Although the supralinear summation of synchronizing excitatory postsynaptic potentials (EPSPs) and backpropagating action potentials (APs) is important for spike-timing-dependent synaptic plasticity (STDP), the spatial conditions of the amplification in the divergent dendritic structure have yet to be analyzed. In the present study, we simulated the coincidence of APs with EPSPs at randomly determined synaptic sites of a morphologically reconstructed hippocampal CA1 pyramidal model neuron and clarified the spatial condition of the amplifying synapses. In the case of uniform conductance inputs, the amplifying synapses were localized in the middle apical dendrites and distal basal dendrites with small diameters, and the ratio of synapses was unexpectedly small: 8-16% in both apical and basal dendrites. This was because the appearance of strong amplification requires the coincidence of both APs of 3-30 mV and EPSPs of over 6 mV, both of which depend on the dendritic location of synaptic sites. We found that the localization of amplifying synapses depends on A-type K+ channel distribution because backpropagating APs depend on the A-type K+ channel distribution, and that the localizations of amplifying synapses were similar within a range of physiological synaptic conductances. We also quantified the spread of membrane amplification in dendrites, indicating that the neighboring synapses can also show the amplification. These findings allowed us to computationally illustrate the spatial localization of synapses for supralinear summation of APs and EPSPs within thin dendritic branches where patch clamp experiments cannot be easily conducted. PMID- 15114050 TI - Extracting wave structure from biological data with application to responses in turtle visual cortex. AB - Waves have long been thought to be a fundamental mechanism for communicating information within a medium and are widely observed in biological systems. However, a quantitative analysis of biological waves is confounded by the variability and complexity of the response. This paper proposes a robust technique for extracting wave structure from experimental data by calculating "wave subspaces" from the KL decomposition of the data set. If a wave subspace contains a substantial portion of the data set energy during a particular time interval, one can deduce the structure of the wave and potentially isolate its information content. This paper uses the wave subspace technique to extract and compare wave structure in data from three different preparations of the turtle visual cortex. The paper demonstrates that wave subspace caricatures from the three cortical preparations have qualitative similarities. In the numerical model, where information about the underlying dynamics is available, wave subspace landmarks are related to activation and changes in behavior of other dynamic variables besides membrane potential. PMID- 15114051 TI - Comparison of coding capabilities of Type I and Type II neurons. AB - We consider the dependence of information transfer by neurons on the Type I vs. Type II classification of their dynamics. Our computational study is based on Type I and II implementations of the Morris-Lecar model. It mainly concerns neurons, such as those in the auditory or electrosensory system, which encode band-limited amplitude modulations of a periodic carrier signal, and which fire at random cycles yet preferred phases of this carrier. We first show that the Morris-Lecar model with additive broadband noise ("synaptic noise") can exhibit such firing patterns with either Type I or II dynamics, with or without amplitude modulations of the carrier. We then compare the encoding of band-limited random amplitude modulations for both dynamical types. The comparison relies on a parameter calibration that closely matches firing rates for both models across a range of parameters. In the absence of synaptic noise, Type I performs slightly better than Type II, and its performance is optimal for perithreshold signals. However, Type II performs well over a slightly larger range of inputs, and this range lies mostly in the subthreshold region. Further, Type II performs marginally better than Type I when synaptic noise, which yields more realistic baseline firing patterns, is present in both models. These results are discussed in terms of the tuning and phase locking properties of the models with deterministic and stochastic inputs. PMID- 15114052 TI - Gene expression profiling: from microarrays to medicine. AB - With the mapping of the human genome comes the ability to identify genes of interest in specific diseases and the pathways involved therein. Laboratory technology has evolved in parallel, providing us with the ability to assay thousands of these genes at once, a technique known as microarray analysis. The main question that this type of technology raises is how we can apply this powerful technology to clinical medicine. Recently, advances in data analysis, as well as standardization of the technology, have allowed us to examine this question, and indeed a few clinical trials currently being performed include microarrays as part of their protocol. In this review we outline the microarray technique and describe these types of studies in further detail. PMID- 15114053 TI - Antibody response to influenza vaccination in splenectomized patients in Poland. AB - The aim of this study was to assess antibody response in 62 splenectomized patients and in 55 healthy subjects vaccinated with split influenza vaccine ("Fluarix," SmithKline Beecham). Response to hemagglutinin and neuraminidase was assessed before vaccination and after 1 month by hemagglutination inhibition test and neuraminidase inhibition test. After vaccination, antibody titers significantly increased in both groups. Postvaccination protection rates ranged from 62.9 to 90.3% in the splenectomized patients and from 81.8 to 94.5% in the control group. Response rates ranged from 50.0 to 75.8% and from 60.0 to 70.9%, respectively. Splenectomized patients produced high antibody levels regardless of the time elapsed from the operation to the vaccination. All requirements of the Committee for Proprietary Medicinal Products regarding humoral response to influenza vaccination in healthy people were fulfilled in both groups. The response in the splenectomized patients was comparable to that in the control group. PMID- 15114054 TI - Essential role of OX40L on B cells in persistent alloantibody production following repeated alloimmunizations. AB - OX40/OX40 ligand (OX40L) interactions are implicated in costimulation for both CD4(+) T and B cells in a bidirectional manner. To determine the role of OX40/OX40L interactions in recipient antidonor responses after multiple allogeneic transfusions, we examined alloreactive cytotoxic T lymphocyte (allo CTL) activity and alloantibody production in repeatedly alloimmunized OX40L deficient mice. After the fifth alloimmunization, whereas OX40L-deficient mice showed allo-CTL activity with levels comparable to those of wild-type mice, alloantibody production in OX40L-deficient mice was significantly reduced, accompanied by fewer memory B and CD4(+) T cells with reduced function. Furthermore, nu/nu mice that received OX40L-deficient T cells still exhibited impaired alloantibody production with fewer memory CD4(+) T and B cells. In contrast, RAG-2-deficient mice that received both wild-type T cells and OX40L deficient B cells produced scant alloantibodies with fewer memory B cells, but sufficient memory CD4(+) T cells. Thus, OX40L on B cells, rather than on T cells, is apparently required for adequate and persistent production of alloantibodies after repeated alloimmunizations. PMID- 15114055 TI - Effect of vitamin D3 on phagocytic potential of macrophages with live Mycobacterium tuberculosis and lymphoproliferative response in pulmonary tuberculosis. AB - Immune responses are elicited through antigen presentation and recognition by macrophages and T-lymphocytes, respectively. The immunomodulatory effect of vitamin D(3) on macrophage phagocytic potential with live Mycobacterium tuberculosis, spontaneous and M. tuberculosis culture filtrate antigen induced lymphocyte responses were studied in pulmonary tuberculosis patients (PTBPs) ( n = 31) and normal healthy subjects (NHSs) ( n = 43). Vitamin D(3) at a concentration of 10(-7) M significantly enhanced the macrophage phagocytosis of live M. tuberculosis in normal subjects with low phagocytic potential (less than 10%) ( p = 0.015). No such increase was observed in PTBPs. Vitamin D(3) significantly decreased the spontaneous lymphoproliferative response ( p = 0.022) and increased the apoptosis of peripheral blood mononuclear cells in PTBPs ( p = 0.024). In normals, vitamin D(3) increased the spontaneous lymphoproliferative response. An inverse correlation between macrophage phagocytosis and spontaneous response was observed in NHSs, whereas a direct correlation was seen between vitamin D(3)-treated cells in normal subjects under in vitro condition. Vitamin D(3) decreased the M. tuberculosis culture filtrate antigen induced lymphocyte response significantly in normal subjects ( p = 0.0003), while it had no influence on the lymphocyte response in PTBPs. The present study suggests that exposure to vitamin D(3) increases the phagocytic potential and spontaneous lymphoproliferative response but brings down the antigen-induced response in normals. In tuberculosis, addition of vitamin D(3) has no significant effect on antigen-induced lymphoproliferative response. This may be due to the unresponsive nature of the cells to the action of vitamin D(3) by virtue of the disease, which renders them inactive. PMID- 15114057 TI - Functional characterization of human Tc0, Tc1 and Tc2 CD8+ T cell clones: control of X4 and R5 HIV strain replication. AB - CD8(+) T lymphocytes have the potential ability to inhibit human immunodeficiency virus (HIV) replication, by secreting soluble(s) factor(s) known as CD8(+) T lymphocyte antiviral factor (CAF). A panel of CD8(+) and CD4(+) T cell clones from HIV1-infected and uninfected donors were generated to better define the phenotype of CAF-producing cells. We first verified that the different CD4(+) T cell subsets (Th0, Th1, and Th2) were productively infected by X4 and R5 virus strains. X4 viral replication in CD4(+) T cells was controlled by the three CD8(+) T cell subsets (Tc0, Tc1, and Tc2); however, the frequency of Tc clones controlling R5 strain was much lower with a dramatic absence of this activity among Tc clones from uninfected donor. Finally, capacity to control viral replication showed an heterogeneity: some clones could control both virus strains, some controlled only the X4 virus, whereas the majority exerted no suppressive activity. PMID- 15114056 TI - Oligoclonal CD8+ T-cell expansion in patients with chronic hepatitis C is associated with liver pathology and poor response to interferon-alpha therapy. AB - The role of CD8(+) T lymphocytes in chronic hepatitis C virus (HCV) infection and in liver injury with subsequent development of fibrosis and cirrhosis is poorly understood. To address this question, we performed a follow-up study including 27 chronically HCV-infected individuals. We determined clonality and phenotypes of circulating CD8(+) T cells employing TCRBV spectratyping. Antigen specificity was tested by rMHC-peptide tetramer staining and stimulation with recombinant HCV antigens. In addition, T-cell clonality and phenotypes were followed during the variable clinical response of interferon- (IFN) alpha treatment. We could demonstrate that CD8(+) T-cell expansions were significantly associated with liver fibrosis and cirrhosis. Likewise, increased oligoclonality of circulating CD8(+) T cells in chronic HCV infection was identified as an indicator for poor clinical response to IFN-alpha therapy. Moreover, we also found that IFN-alpha therapy enhanced the differentiation of CD8(+) T cells towards a late differentiation phenotype (CD28(-) CD57(+)). In cases of virus elimination the disappearance of expanded terminally differentiated CD8(+) cells was observed. Thus, this study identifies an association of clonal expansions of circulating CD8(+) T cells with liver pathology and provides a possible explanation for the fact that response to IFN-alpha therapy diminishes with the duration of infection. PMID- 15114058 TI - Altered coexpression of lectin-like receptors CD94 and CD161 on NK and T cells in HIV patients. AB - HIV patients either on highly active antiviral therapy (HAART) or therapy naive were analyzed for their CD56 phenotype and cytokine production in comparison to healthy controls (HC). Both NK cell populations (CD56(dim) and CD56(bright)) are found to be present in all groups with selectively decreased CD56(dim) NK cells in HAART naive patients. Patients on HAART exhibited significantly diminished numbers of CD161+CD56(dim) NK cells. CD56(dim) were equally potent in producing IFNgamma in all three groups. The number of TNFalpha+CD56(bright) NK cells from patients on HAART and TNFalpha+CD56(dim) NK cells from HAART naive patients was significantly reduced as compared to healthy controls. In summary our data revealed that functional capacities and coexpression patterns of lectin-like receptors on lymphocytes are differentially affected in HIV patients depending on the state of therapy (under HAART or HAART naive) or the cell type (NK or T cells), respectively. PMID- 15114059 TI - Leptin enhances the release of cytokines by peripheral blood mononuclear cells from relapsing multiple sclerosis patients. AB - Leptin, a hormone synthesized mainly by adipocytes, can modulate the immune response and seems to be involved in the induction of experimental autoimmune encephalomyelitis, an animal model of multiple sclerosis (MS). However, the possible role of leptin in MS pathogenesis has not yet been elucidated. In this study we investigated the effect of leptin on cytokine production by peripheral blood mononuclear cells (PBMCs) of MS patients (either in the acute or in the stable phase of the disease) and healthy controls. We also analyzed leptin effects on cytokine production by monocytes in relapsing MS patients. Our data showed that leptin induced tumor necrosis factor-alpha, interleukin-6, and interleukin-10 production by PBMCs of patients in an acute phase of disease but not in patients in a stable phase or in healthy controls. Moreover, we found no effect of leptin in monocytes from relapsing MS patients. Therefore we conclude that leptin may modulate the MS inflammatory process during relapses. PMID- 15114060 TI - A novel approach for cancer immunotherapy: tumor cells with anchored superantigen SEA generate effective antitumor immunity. AB - Murine B16 melanoma cell line is poorly immunogenic and highly aggressive. We recently reported that the transmembrane staphylococcal enterotoxin A (TM-SEA) anchors onto B16 cells and stimulates lymphocyte proliferation. The purpose of the study was to investigate whether vaccination with B16 cells bearing membrane anchored TM-SEA fusion protein could cause tumor-specific immunity. Mice in the therapeutic vaccination group received B16 tumor inoculations, followed by treatment with B16-TM-SEA vaccine or control vaccines. Mice in the prophylactic vaccination group were given B16-TM-SEA vaccine or control vaccines, followed by challenge with wild type B16 or control EL4 cells. Significant tumor growth inhibition, prolongation of survival, and marked augmentation of NK and CTL activities were observed in mice which received B16-TM-SEA vaccine as compared to controls. Overall, our results suggest that the TM-SEA cellular vaccine is a novel and effective strategy for cancer immunotherapy. PMID- 15114062 TI - Octagam 5%, an intravenous IgG product, is efficacious and well tolerated in subjects with primary immunodeficiency diseases. AB - Octagam is an intravenous immunoglobulin preparation registered in Europe for treating primary immunodeficiency diseases (PID). The present clinical trial was designed to demonstrate that Octagam meets the minimal efficacy requirement of the U.S. Food and Drug Administration-that treatment should result in 0.15). The incidence of acute rejection was significantly lower during the first 2 months posttransplantation (P =0.002) and slightly lower overall in the study group versus the control group at 12 months (46% vs. 55%, P =0.12, log-rank test). Median time to rejection among those experiencing rejection was significantly longer in the study group versus control group (2.76 vs. 0.34 months, P =0.0007). The mean Tac dose, 12-hr trough level, and percentage of patients receiving maintenance steroids were significantly lower in the group receiving C1H and Tac (P <0.0001 during the first 3 months, P <0.05 thereafter), as were the mean creatinine levels (P <0.05) and incidence of nephrotoxicity (P =0.004, conversion from Tac to other agents). Finally, in the group receiving C1H/Tac, patients with an average Tac trough level less than 6.5 ng/mL during the first 2 months post-transplantation demonstrated a significantly higher rejection rate beyond that time (P =0.02). CONCLUSION: C1H and low-dose Tac seems to be at least as effective as our standard Tac and steroids regimen in preventing acute rejection in adult liver allotransplantation with less renal toxicity and less use of maintenance steroids. PMID- 15114088 TI - Sirolimus-associated pulmonary toxicity. AB - BACKGROUND: Pulmonary toxicity has recently been recognized as a potentially serious complication associated with sirolimus therapy. We further detail this condition on the basis of our own cases and those reported in the literature. METHODS: We report three cases of suspected sirolimus-induced pulmonary toxicity that occurred in three renal transplant recipients and searched PubMed for all previously reported cases. RESULTS: Including our current cases, 43 patients with sirolimus-induced pulmonary toxicity have now been reported. Clinical data were incomplete in 28 cases. Analysis of available data for 15 patients revealed that the most commonly presenting symptoms were dyspnea on exertion and dry cough followed by fatigue and fever. Chest radiographs and high-resolution computed tomography scans commonly revealed bilateral patchy or diffuse alveolo interstitial infiltrates. Bronchoalveolar fluid analysis and lung biopsy in selected case reports revealed several distinct histologic features, including lymphocytic alveolitis, lymphocytic interstitial pneumonitis, bronchoalveolar obliterans organizing pneumonia, focal fibrosis, pulmonary alveolar hemorrhage, or a combination thereof. The diagnosis of sirolimus-associated pulmonary toxicity was made after an exhaustive work-up to exclude infectious causes and other pulmonary disease. Sirolimus discontinuation or dose reduction resulted in clinical and radiologic improvement in all 15 patients within 3 weeks. CONCLUSION: The temporal relationship between sirolimus exposure and onset of pulmonary symptoms in the absence of infectious causes and other alternative pulmonary disease and the associated clinical and radiologic improvement after its cessation suggests a causal relationship. Because the use of sirolimus in organ transplantation has become more widespread, clinicians must remain vigilant to its potential pulmonary complication. PMID- 15114089 TI - Efficacy and safety of tacrolimus compared with cyclosporine microemulsion in primary simultaneous pancreas-kidney transplantation: 1-year results of a large multicenter trial. AB - BACKGROUND: Simultaneous pancreas-kidney transplantation (SPK) transplantation has become an accepted therapy for type 1 diabetic patients with end-stage renal disease. This open-label, multicenter study compared the efficacy and safety of tacrolimus with the microemulsion (ME) formulation of cyclosporine in a clinical setting. The 1-year results are reported here. METHODS: The study was conducted in 10 European centers and one center in Israel. One hundred three patients were randomly assigned to tacrolimus and 102 to cyclosporine-ME. All patients received concomitant rabbit anti-T-cell globulin induction therapy, mycophenolate mofetil (MMF), and short-term cortico-steroids. The initial daily oral doses were 0.2 mg/kg for tacrolimus, 7 mg/kg for cyclosporine-ME, and 2 to 3 g for MMF. RESULTS: The 1-year incidence of biopsy-proven kidney or pancreas acute rejection was lower with tacrolimus (27.2%) than with cyclosporine-ME (38.2%; P = 0.09). Pancreas graft survival at 1 year was 91.3% with tacrolimus and 74.5% with cyclosporine-ME (P <0.0005). Renal graft survival was similar in the two study groups. There were no significant treatment-related differences in pancreatic or renal graft function. In total, 34 patients switched treatment from cyclosporine ME to tacrolimus, but only 6 patients receiving tacrolimus required alternative therapy. Mean doses of MMF at 1 year were also lower in the tacrolimus group (1.36 vs. 1.67 g/day; P = 0.007). CONCLUSION: These findings support the use of tacrolimus therapy for uremic patients with type 1 diabetes who are undergoing SPK transplantation. PMID- 15114090 TI - Comparison of sirolimus-based calcineurin inhibitor-sparing and calcineurin inhibitor-free regimens in cadaveric renal transplantation. AB - INTRODUCTION: This study examines the efficacy and toxicity of sirolimus used as primary immunosuppression in combination with reduced dose tacrolimus (calcineurin inhibitor [CI]-sparing regimen) or mycophenolate mofetil (CI-free regimen) in high-risk cadaveric renal transplantation. METHODS: Seventy subjects were treated in a quadruple sequential protocol in which 41 were treated with a CI-sparing regimen and 29 were treated with a CI-free regimen. The efficacy and toxicity profiles of these regimens were prospectively monitored and compared. RESULTS: The study consisted of African Americans (71%), cadaveric donors (100%), donors aged more than 50 years (30%), and patients with delayed graft function (47%). At 1 year, patient survival, graft survival, and incidence of biopsy proven acute rejection were 98%, 80%, and 10%, respectively, in the CI-sparing group and 100%, 89%, and 7%, respectively, in the CI-free group. Three-month protocol biopsies were performed in 41% (17/41) and 67% (20/29) of the subjects in the CI-sparing and CI-free groups, respectively. Subclinical rejection was detected in 6% (1/17) and 15% (3/20) of the subjects in the CI-sparing and CI free groups, respectively. Histologic evidence of chronic allograft nephropathy was more prevalent in the CI-sparing group. At 1 year, the mean estimated creatinine clearance was higher in the CI-free group than in the CI-sparing group (72.4 +/-20.0 mL/min vs. 50.5 +/-20.8 mL/min, P <0.01). The two regimens had similar toxicity profiles (hospital readmission, infection, wound complications, and metabolic complications). CONCLUSIONS: Both sirolimus-based CI-sparing and CI free regimens are safe and effective in a population with high immunologic risk. The CI-free regimen is associated with better renal function at 1 year post transplant. Long-term follow-up will aid in determining the risk and benefit ratio of these regimens. PMID- 15114091 TI - The number of amino acid triplet differences between patient and donor is predictive for the antibody reactivity against mismatched human leukocyte antigens. AB - BACKGROUND: The correlation between antibody production against mismatched donor human leukocyte antigens (HLA) and the number of amino acid sequence mismatches was analyzed in patients who rejected a kidney transplant (n=146). METHODS: A similar analysis was performed for the antibody production of women against the paternal HLA antigens of their child (n=1,397). The amino acid sequence (triplet) differences were analyzed using the HLAMatchmaker algorithm. RESULTS: In both groups, a positive correlation was found between the number of triplet mismatches and the percentage of individuals producing antibodies (P <0.0001). If zero triplet mismatches were present, no antibodies were formed in all cases. When 11 or 12 triplet mismatches were present, 94% of the transplant patients produced antibodies against the donor. In pregnancy, 11 or 12 triplet mismatches led to 27% of the women producing specific antibodies. CONCLUSIONS: These results indicate that the immunogenicity of the fetus is lower than that of a rejected kidney and that analysis of the number of triplet mismatches can predict the antibody reactivity against the mismatched HLA antigens. PMID- 15114092 TI - Xanthine oxidoreductase and preservation injury in human liver transplantation. AB - BACKGROUND: Preservation injury is a major cause of primary graft dysfunction in liver transplantation (LT). Oxidative damage is considered to be the first event leading to graft damage. Xanthine oxidoreductase (XOR) and neutrophil activation, two sources of reactive oxygen species, could play a role in the development of graft dysfunction. METHODS: We determined activities of XOR forms, polymorphonuclear elastase (PMN-E), aminotransferases, and hyaluronic acid in plasma of 20 patients undergoing LT. Samples were taken from the radial artery (RA) before the anhepatic phase; from the portal vein (PV) before reperfusion; from graft caval effluent (CE) at reperfusion; and from RA, PV, and the hepatic vein (HV) 10 and 90 min postreperfusion. RESULTS: The graft, but not recipient bowel, released XOR into blood (XOR in CE, median, 61.2 mU/g protein [range, 1.9 160.4 vs. undetectable in PV before reperfusion). Circulating XOR was transformed from dehydrogenase to reversible oxidase (XOrev) (XOrev-to-XOR ratio, 48.1% in CE and 65.1% in HV 90 min postreperfusion). Neutrophil activation was detected in the recipients before reperfusion, and in liver at early post-reperfusion (median PMN-E was 0.85 microg/g protein [range, 0.01-1.58] in RA before the anhepatic phase; 2.22 microg/g protein [range, 0.20-5.88] in PV prereperfu-sion; and 3.60 microg/g protein [range, 0.48-6.78] in HV 10 min postreperfusion). XOR, but none of the other markers, was higher in the CE of patients with moderate primary graft dysfunction than in those with slight primary graft dysfunction. CONCLUSIONS: XOR release and neutrophil activation are produced during LT, and they are potentially injurious mechanisms associated with this therapy. PMID- 15114093 TI - Demonstration that donor-specific nonresponsiveness in human liver allograft recipients is both rare and transient. AB - BACKGROUND: The side effects of lifetime immunosuppression are a major cause of morbidity and mortality; however, in the absence of prospective monitoring, immunosuppression withdrawal may lead to graft loss from rejection. To detect and monitor suitable recipients for immunosuppression withdrawal, the authors used an in vitro assay of T-cell function to study 71 long-term liver allograft recipients. METHODS: Interleukin-2 secretion by blood mononuclear cells was measured in response to recall antigens, alloantigen (donor and third-party), and phytohemagglutinin. RESULTS: Forty-four recipients were studied at a single time point at least 1 month after transplantation. The majority reacted to all antigens (n=33), whereas four showed globally reduced or absent responses (n=4) and six had markedly reduced or absent responses to donor alloantigen in the presence of preserved responses to third-party alloantigen and recall antigens. Four of these donor-nonresponsive recipients were retested 6 to 12 months later, by which time all had redeveloped responses to donor alloantigen. Serial measurements for up to 2 years in a prospective cohort of 27 liver allograft recipients showed only two patients to be consistently donor-nonresponsive posttransplant. CONCLUSIONS: Most patients rapidly reacquire vigorous immune responses after liver transplantation, and only a minority are hyporesponsive to donor alloantigen. Donor-specific nonresponsiveness is transient in most patients, and serial monitoring is required to define sustained periods of donor specific nonresponsiveness. Whether such patients are suitable for immunosuppressive withdrawal is unclear. PMID- 15114094 TI - Hepatitic graft-versus-host disease after hematopoietic stem cell transplantation: clinicopathologic features and prognostic implication. AB - BACKGROUND: Graft-versus-host disease (GVHD) of the liver after allogeneic hematopoietic stem cell transplantation classically presents with increased bilirubin and alkaline phosphatase (ALP) levels. A hepatitic variant was recently recognized, with more than a 10-fold increase in aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. This study defines the clinicopathologic features and prognostic implications of hepatitic GVHD compared with classic liver GVHD. METHOD: A total of 38 cases of hepatitic GVHD, 68 cases of classic liver GVHD, and 13 cases of hepatitis B virus (HBV)-related hepatitis after hematopoietic stem cell transplantation were analyzed. RESULTS: Hepatitic GVHD cases showed significantly higher ALT, AST, and ALP levels compared with classic liver GVHD cases (at onset, mean ALT: 154 vs. 58 U/L, P <0.001; AST: 167 vs. 77 U/L, P <0.001; at peak, ALT: 435 vs. 112 U/L, P <0.001; AST: 587 vs. 150 U/L, P <0.001; ALP: 416 vs. 238 U/L, P =0.001), persisted longer (74 vs. 32 days, P =0.006), and showed more lobular pathologic changes in biopsy (lobular changes: 16/26 vs. 4/19, P =0.007; hepatocyte necrosis: 16/26 vs. 6/19, P =0.008; acidophil bodies: 15/26 vs. 4/19, P =0.014) but less cholestasis (4/26 vs. 8/19, P =0.045). However, cumulative doses of immunosuppressants prescribed, response, and outcome were similar. Compared with hepatitic GVHD, HBV-related hepatitis occurred later (95 vs. 184 days, P =0.049), but clinical and biochemical profiles were similar, requiring liver biopsies for their distinction. CONCLUSIONS: Hepatitic and classic liver GVHD differed biochemically and pathologically, but these differences showed no obvious impact on outcome. The distinction of hepatitic GVHD from other hepatitis is mandatory. PMID- 15114095 TI - Detection of Aspergillus galactomannan by enzyme immunoabsorbent assay in recipients of allogeneic hematopoietic stem cell transplantation: a prospective study. AB - BACKGROUND: Invasive aspergillosis (IA) has become the leading infectious cause of death after allogeneic hematopoietic stem cell transplantation (allo-HSCT). This is partially because of the lack of a sensitive, specific, and noninvasive diagnostic test. New diagnostic tests for IA, such as the detection of Aspergillus galactomannan antigen (AGA) by sandwich enzyme-linked immunoabsorbent assay (ELISA), have recently been described. This study validates the usefulness of this diagnostic tool in the allo-HSCT setting. METHODS: From January 1999 to January 2001, all consecutive adult patients undergoing allo-HSCT were prospectively studied with a galactomannan antigenemia assay (ELISA test) twice weekly from admission until death or discharge, and weekly afterward if the patient received immunosuppressive therapy. Proven, probable, and possible IA were defined according to the European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria. RESULTS: During the 2 years of study, 74 patients underwent an allo-HSCT. A total of 832 serum samples were collected. According to the European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria, it was ascertained that 66 patients did not fulfill any criteria of IA, 2 patients were classified with possible IA, 5 patients were classified with probable IA, and 1 patient was classified with proven IA. Fourteen samples were positive for AGA, all from patients with IA. The sensitivity and specificity of the test were 75% and 100%, respectively. The positive predictive and negative predictive values were 100% and 97%, respectively. CONCLUSIONS: In this study, AGA detection was clearly related to IA. Although the ELISA test did not have any role in the anticipation of the diagnosis, it clarifies the diagnosis of IA in allo-HSCT. PMID- 15114096 TI - Pulsatile machine perfusion with Vasosol solution improves early graft function after cadaveric renal transplantation. AB - BACKGROUND: Cold storage is the most common method of preservation in renal transplantation, but pulsatile machine perfusion (MP) is increasingly used for purposes of ex vivo assessment and resuscitation of high risk allografts. Vasosol (VSL) MP solution (MPS) (Pike Laboratories Inc, Eagle, PA) is a novel perfusate with enhanced vasodilatory and antioxidant capacity. We report our experience with VSL in machine preservation of renal allografts. METHODS: Locally procured cadaver kidney pairs undergoing MP were randomized to VSL or control solution (Belzer MPS; Trans-Med, Elk River, MN). En bloc perfusion was performed according to standard MP procedures. Transplantation was performed at 1 of 12 local transplant centers. Donor and recipient data were collected prospectively. RESULTS: Data from 162 transplanted kidneys were analyzed. A total of 82 renal grafts were perfused with VSL, and 80 were perfused with control solution. No organs were lost secondary to the technique of MP. There was no difference in donor or recipient age between groups. Kidneys perfused with VSL exhibited significantly higher rates of immediate function and significantly less delayed graft function (12.2% vs. 21.2%). Discharge creatinine and length of stay were also significantly improved in the VSL group. One-year graft and patient survivals were equivalent (95%) in both groups. CONCLUSIONS: VSL improved early graft function and shortened the length of stay compared with Belzer MPS. Further improvement in preservation solutions in conjunction with pulsatile perfusion shows promise in improving early outcomes after renal transplantation, especially for extended criteria donor kidneys. PMID- 15114097 TI - Prospective, randomized trial of the effect of antibody induction in simultaneous pancreas and kidney transplantation: three-year results. AB - BACKGROUND: Historically, antibody induction has been used because of the higher immunologic risk of graft loss or rejection observed in simultaneous pancreas and kidney (SPK) transplantation compared with kidney transplantation alone. This trial was designed to assess the effect of antibody induction in SPK transplant recipients receiving tacrolimus, mycophenolate mofetil, and corticosteroids. Induction agents included T-cell-depleting and interleukin-2 receptor antibodies. METHODS: A total of 174 SPK transplant recipients were enrolled in a prospective, open-label, multi-center study. They were randomized to induction (n=87) or non induction (n=87) groups and followed for 3 years. RESULTS: At 3 years, actual patient (94.3% and 89.7%) and pancreas (75.9% and 75.9%) survivals were similar between the induction and non-induction groups, respectively. Actual kidney survival was similar at 1 and 2 years, but at 3 years, it was significantly better in the induction group compared with the non-induction group (92% vs. 81.6%; P =0.04). At 3 years, median serum creatinine and hemoglobin A1C were similar between the induction and non-induction groups (1.35 mg/dL and 1.20 mg/dL, 5.4% and 5.5%, respectively). Three-year cumulative incidence of biopsy confirmed, treated acute kidney rejection in the induction and non-induction groups was 19.5% and 27.5% (P =0.14), respectively, with odds 4.6 times greater in African Americans regardless of treatment (P =0.004). Significantly higher rates of cytomegalovirus (CMV) viremia and CMV syndrome occurred in those receiving T-cell-depleting antibody induction (36.1%) when compared with those receiving anti-interleukin-2 receptor antibodies (2%) and non-induction (8.1%) (P <0.0001). CONCLUSIONS: Tacrolimus, mycophenolate mofetil, and corticosteroids resulted in excellent safety and efficacy in SPK transplant recipients. Actual 3 year kidney survival was significantly better in the induction group; however, CMV viremia and CMV syndrome rates were significantly higher in the T-cell depleting antibody group. African Americans demonstrated a significantly greater risk of acute rejection despite antibody induction. Decisions regarding the use of induction therapy must weigh the risk of kidney graft loss or rejection against the risk of infection. PMID- 15114098 TI - Importance of the Gal alpha1-3 Gal antigen in discordant islet xenotransplantation: immunosuppression, which inhibits porcine islet xenograft rejection in ordinary mice, is equally effective in Gal-knockout mice. AB - BACKGROUND: Islet xenotransplantation will most likely be performed in diabetic patients treated with immunosuppressive drugs. The importance of the galactosyl alpha(1-3) galactose (Galalpha1-3Gal) antigen in immunosuppressed islet xenograft recipients has not been studied. METHODS: Fetal porcine islet-like cell clusters (ICCs) were transplanted into the renal subcapsular space of both Gal-knockout mice and ordinary mice. Transplantations were performed in untreated mice and mice immunosuppressed with cyclosporine A (CsA) plus 15-deoxyspergualin (DSG). Studies were also performed in immunosuppressed Gal-knockout mice that had been actively immunized against Galalpha1-3Gal. Evaluation was performed 12 days after transplantation using morphologic techniques. The levels of serum immunoglobulin (Ig)G and IgM to the Galalpha1-3Gal antigen or to the ICCs were determined. RESULTS: No difference in the morphologic appearance could be seen between ordinary mice and Gal-knockout mice. No deposits of IgG, IgM, or C3 could be detected. Almost no difference could be seen between immunosuppressed Gal knockout mice and immunosuppressed ordinary mice. In immunosuppressed, immunized Gal-knockout mice, the results were similar. In ordinary mice treated with CsA+DSG, the levels of anti-Gal IgM were lower than they were in untreated mice, whereas the levels of anti-Gal IgG were similar. In Gal-knockout mice (including immunized animals) treated with CsA+DSG, the levels of anti-Gal IgG and IgM were lower than they were in untreated Gal-knockout mice. CONCLUSIONS: After renal subcapsular transplantation, antibodies against Galalpha1-3Gal have no major influence on islet xenograft rejection in the pig-to-mouse model. Immunosuppression, which inhibits rejection in the pig-to-mouse model, is equally effective when transplantation is performed across the Galalpha1-3Gal barrier. PMID- 15114099 TI - Effect of tacrolimus on the expression of macrophage scavenger and nuclear hormone receptors in THP-1-derived human macrophages. AB - BACKGROUND: Data indicate that tacrolimus and cyclosporine A (CsA) differentially affect the risk of atherosclerosis. The results of our recent in vitro studies of clinically relevant CsA concentrations demonstrated the modulation of macrophage scavenger receptors (MSRs) involved in atherogenesis. This work evaluated the effects of clinically relevant tacrolimus concentrations on the expression of the MSR genes CD36 and CD68, SR-A and SR-BII, lectin-like oxidized low-density lipoprotein receptor-1, the nuclear hormone receptors peroxisome proliferator activated receptor (PPAR)gamma and liver-X-receptor-alpha, and the cholesterol efflux pump ABCA1 in the in vitro human THP-1 macrophage model. METHODS: The cells were cultured and differentiated into macrophages. Macrophages were treated with the tacrolimus to assess gene expression in a time-dependent (1, 2, 4, 8, and 24 hr) and dose-dependent (concentrations [micrograms/liter] corresponding to the trough [15], peak [30], and 4 x peak [120]) manner using reverse transcriptase polymerase chain reactions. The gene expression levels of interest were normalized to GAPDH expression in each sample to provide semiquantitative reverse-transcriptase polymerase chain reaction results. Additional immunoblotting studies demonstrated protein expression of CD36, PPARgamma, and ABCA1. RESULTS.: The gene expression of CD36, SR-BII, and lectin-like oxidized low-density lipoprotein receptor-1 were down-regulated, and ABCA1 was up regulated. CD68, SR-AI, liver-X-receptor-alpha, and PPARgamma were regulated in a dose-dependent manner. Protein expression of CD36 was down-regulated, and PPARgamma and ABCA1 were relatively unchanged. CONCLUSIONS: Tacrolimus seems to regulate MSRs, nuclear hormone receptors, and ABCA1 in THP-1 macrophages. These results differ from previous findings with CsA and may provide insight into the mechanisms of posttransplant atherosclerosis. PMID- 15114100 TI - A novel monoclonal antibody inhibits the immune response of human cells against porcine cells: identification of a porcine antigen homologous to CD58. AB - BACKGROUND: Human CD58 is an adhesion molecule that interacts with CD2 on lymphocytes. We describe here an antibody that blocks responses of human peripheral blood mononuclear cells (PBMCs) to porcine cells and reacts with a porcine protein with homology to CD58. METHODS: Antibodies were isolated with a screen for inhibition of the human antiporcine response. One of these antibodies was used for immunoaffinity purification of a protein that was identified by molecular weight determination, endoglycosidase sensitivity, and microsequencing analysis as a porcine homologue of CD58. RESULTS: The antigen recognized by this antibody was a cell surface protein of relative molecular mass (Mr)=45,000 containing N-linked carbohydrate chains. Immunoaffinity purification of this protein and microsequencing revealed homology to sheep CD58 as well as sequences that were common to this protein and both sheep and human CD58. The protein was widely distributed on porcine cells, including lymphocytes, endothelial cells, muscle cells, and neuronal cells. This antibody efficiently inhibited lysis of porcine targets by human PBMCs in addition to preventing proliferation of the human PBMCs in response to the porcine cells. CONCLUSIONS: The CD2 interaction with porcine cells is important for the efficient recognition of porcine tissue, and inhibition of the human antiporcine immune response with the antibody is likely to be caused by the disruption of the human CD2 interaction with this porcine homologue of CD58. The antibody may prove to be useful for the blocking of this interaction without interfering with other functions of T cells. PMID- 15114101 TI - Anti-tubular basement membrane antibodies and giant cell formation in a model of chronic renal allograft rejection. AB - BACKGROUND: In the Fisher (F344)-to-Lewis (LEW) kidney transplantation (Tx) model of chronic rejection, antibodies reactive with tubular basement membranes have been found. We investigated whether giant cells, typical for a granulomatous inflammatory reaction, can be found during allograft rejection. METHODS: F344-to LEW renal Txs were performed, and kidneys were removed at various time points. Kidneys were analyzed by histology and immunohistochemistry. Interferon-gamma levels were measured by reverse-transcriptase polymerase chain reaction. RESULTS: Multinucleated Langhans'-type giant cells were found in F344 allografts at days 21 and 30 and disappeared thereafter. Giant cells were not observed in LEW allografts. The giant cells were ED1, OX-43, and major histocompatibility complex class II positive. All grafts showed transient expression of interferon-gamma, peaking at days 7 to 14 post-Tx. CONCLUSION: Granulomatous inflammation with giant cell formation occurs in the F344-to-LEW model and may represent an alloimmune response against tissue-specific antigens. PMID- 15114102 TI - Maturity-onset diabetes of the young with end-stage nephropathy: a new indication for simultaneous pancreas and kidney transplantation? AB - BACKGROUND AND CASE: Simultaneous pancreas and kidney transplantation (SPK) is applied almost exclusively in C-peptide-negative type 1 diabetic patients, although some data on SPK in type 2 diabetes have been published as well. Nothing is known about SPK in the autosomal diabetes form, maturity-onset diabetes of the young (MODY). SPK was performed in a 47-year old man who has MODY3 because of a Arg272His mutation in the hepatocyte nuclear factor-1alphagene. He developed overt diabetes mellitus at 19 years and end-stage diabetic nephropathy 26 years thereafter. Before SPK, the patient had measurable fasting serum C-peptide levels, but lacked beta-cell response to intravenous glucose and glucagon. He was treated with 34 IU of insulin per day. At 2 years post-transplantation, the patient remains normoglycemic and insulin independent. A hyperglycemic clamp test showed a normal beta-cell function. CONCLUSION: Identification of MODY3 among all C-peptide-positive patients with advanced diabetic nephropathy might help to select a specific group profiting from SPK. PMID- 15114103 TI - Influence of islet transportation on pancreatic islet allotransplantation in type 1 diabetic patients within the Swiss-French GRAGIL network. AB - BACKGROUND: The influence of islet transportation on pancreatic islet allotransplantation in type 1 diabetic patients was evaluated within the GRAGIL network. PATIENTS AND METHODS: From December 2001 to April 2003, 16 human pancreatic islet transplants were performed in 9 type 1 diabetic patients with an established kidney graft (functioning for at least 6 months) in four centers of the GRAGIL network. Islet isolation was performed in a core laboratory in Geneva, and the islet preparations were shipped by ambulance to each center for transplantation. One month after transplantation, the efficiency of the graft was assessed according to islet transportation time (ITT): ITT less than 2 hours (group 1, n=5), and ITT greater than 4.5 hours (group 2, n=4, mediant 5 hours). RESULTS: Primary graft dysfunction was observed in one patient in group 1 after one month. Two patients became insulin independent in groups 1 and 2. All other patients in both groups had a plasma C-peptide level greater than 0.5 ng/ml. The HbA1c level and the exogenous insulin needs decreased in both groups. CONCLUSIONS: ITT does not seem to influence the efficiency of pancreatic islet allotransplantation in type 1 diabetic patients. These results emphasize the scope for multicenter networks such as the GRAGIL group. PMID- 15114104 TI - Comparison of histidine-tryptophan-ketoglutarate solution and University of Wisconsin solution for organ preservation in clinical pancreas transplantation. AB - BACKGROUND: University of Wisconsin (UW) solution is currently the standard preservation solution used for abdominal organ transplantation. This study assesses the efficacy of histidine-tryptophan-ketoglutarate (HTK) compared with UW in pancreas transplantation. METHODS: Between October 2002 and August 2003, 20 pancreas transplants were performed. Patients were divided into two groups: UW (n=10) and HTK (n=10). Donor and recipient demographics were similar in both groups. The mean cold ischemia time for both groups was 11 +/-3 hr. RESULTS: There was an anticipated difference between total preservative volumes used (HTK: 4.5 +/- 1.2 L vs. UW: 3.4 +/-0.8 L; P =0.03). Patient and graft survivals to date were 100% in both groups. Serum fasting blood glucose, peak amylase, and serial amylase levels remained comparable at all intervals posttransplantation. CONCLUSIONS: Within this range of cold ischemia time, UW and HTK demonstrate similar efficacy in pancreas preservation. PMID- 15114105 TI - Successful treatment of recurrent cholangitis after adult liver transplantation with a Tsuchida antireflux valve. PMID- 15114106 TI - Quality control of the OPTN/UNOS Transplant Registry. PMID- 15114108 TI - Reconstruction of the hepatic artery in liver transplantation in mice: mandatory or useless? PMID- 15114112 TI - Muscle reconstruction in cleft lip repair. AB - This article presents a method of muscle reconstruction in the cleft lip patient that grossly reproduces the interlacing muscle anatomy in the central portion of the upper lip and thereby addresses both the static and the functional requirements of cleft lip repair. This lip repair procedure is combined with paranasal muscle reconstruction, preoperative nasoalveolar molding, and postoperative nasal stenting to constitute a comprehensive approach to cleft lip/nose repair. PMID- 15114113 TI - Epidemiology of cleft lip and cleft palate in Pakistan. AB - Clinical and epidemiologic studies of defined geographic populations can serve as a means of establishing data important for the diagnosis, treatment, and counseling of patients with cleft lip and cleft palate. Several descriptive epidemiologic studies have been carried out in many countries worldwide; however, no such study has ever been performed in Pakistan. Population-based data on the incidence of cleft lip and palate were obtained from birth registry information in northern Pakistan. A total of 117 cases from 61,156 live births reported were identified. The incidence for cleft lip and/or cleft palate was 1.91 per 1000 births (one per 523 births). Cleft lip alone (42 percent) was noted more frequently than isolated cleft palate (24 percent) and combined cleft lip and palate deformities (34 percent). Boys were more commonly affected by cleft lip and cleft lip with cleft palate, whereas girls predominated in the isolated cleft palate cases. Consanguineous marriages were observed in 32 percent of parents versus 18 percent in matched controls. Only 32 percent of cleft mothers received formal prenatal counseling, monthly examinations, and regular laboratory testing during the entirety of the pregnancy. Nutritional and vitamin supplements were given to only 28 percent of mothers of cleft children versus 59 percent in matched controls. Descriptive statistics were used to assess pertinent risk factors associated with cleft lip and palate. The acquisition of incidence and associated data has generated baseline information on the magnitude of cleft lip and cleft palate in Pakistan. It is hoped that this information can be used for appropriate resource use, cleft lip and cleft palate prevention programs, and counseling programs with Pakistan-specific data. PMID- 15114114 TI - Craniofacial anomalies of the amniotic band syndrome in serial clinical cases. AB - Amniotic band syndrome has been proposed as a sequela of intrauterine rupture of the amnion, resulting in oligohydramnios and passage of the fetus into the chorionic cavity. Amnion disruption with loss of amniotic fluid, causing fetal compression and localized fetal ischemia, possibly results in a pathogenic mechanism of extremely variable malformations. The prominence of the nasal processes and the adjacent stomodeal orifice facilitates free band attachment and adherence, resulting in a spectrum of similarly oriented facial defects. The authors present six consecutive cases of amniotic band syndrome with cleft lip and palate (facial cleft 3, 5, 7, and 10, isolated or combined) that were associated with other craniofacial anomalies, such as craniosynostosis and hypertelorbitism. They also present limb malformations and discuss the proposed pathogenesis and the surgical challenges in functional and aesthetic restoration. PMID- 15114115 TI - Neurovascular free-muscle transfer for the treatment of established facial paralysis following ablative surgery in the parotid region. AB - Neurovascular free-muscle transfer for facial reanimation was performed as a secondary reconstructive procedure for 45 patients with facial paralysis resulting from ablative surgery in the parotid region. This intervention differs from neurovascular free-muscle transfer for treatment of established facial paralysis resulting from conditions such as congenital dysfunction, unresolved Bell palsy, Hunt syndrome, or intracranial morbidity, with difficulties including selection of recipient vessels and nerves, and requirements for soft-tissue augmentation. This article describes the authors' operative procedure for neurovascular free-muscle transfer after ablative surgery in the parotid region. Gracilis muscle (n = 24) or latissimus dorsi muscle (n = 21) was used for transfer. With gracilis transfer, recipient vessels comprised the superficial temporal vessels in 12 patients and the facial vessels in 12. For latissimus dorsi transfer, recipient vessels comprised the facial vessels in 16 patients and the superior thyroid artery and superior thyroid or internal jugular vein in four. Facial vessels on the contralateral side were used with interpositional graft of radial vessels in the remaining patient with latissimus dorsi transfer. Cross-face nerve grafting was performed before muscle transfer in 22 patients undergoing gracilis transfer. In the remaining two gracilis patients, the ipsilateral facial nerve stump was used as the primary recipient nerve. Dermal fat flap overlying the gracilis muscle was used for cheek augmentation in one patient. In the other 23 patients, only the gracilis muscle was used. With latissimus dorsi transfer, the ipsilateral facial nerve stump was used as the recipient nerve in three patients, and a cross-face nerve graft was selected as the recipient nerve in six. The contralateral facial nerve was selected as the recipient nerve in 12 patients, and a thoracodorsal nerve from the latissimus dorsi muscle segment was crossed through the upper lip to the primary recipient branches. A soft-tissue flap was transferred simultaneously with the latissimus muscle segment in three patients. Contraction of grafted muscle was not observed in two patients with gracilis transfer and in three patients with latissimus dorsi transfer. In one patient with gracilis transfer and one patient with latissimus dorsi transfer, acquired muscle contraction was excessive, resulting in unnatural smile animation. The recipient nerves for both of these patients were the ipsilateral facial nerve stumps, which were dissected by opening the facial nerve canal in the mastoid process. From the standpoint of operative technique, the one-stage transfer for latissimus dorsi muscle appears superior. Namely, a combined soft-tissue flap can provide sufficient augmentation for depression of the parotid region following wide resection. A long vascular stalk of thoracodorsal vessels is also useful for anastomosis, with recipient vessels available after extensive ablation and neck dissection. PMID- 15114116 TI - Psychological sequelae of failed scalp replantation. AB - Published reports of avulsed scalp replant attempts have been promising. Numerous case reports and published series have demonstrated a greater than 90 percent replantation success rate. However, there exists a paucity of articles on the management of patients following failed scalp replantation attempts. The authors recognize numerous stressors that affect these patients, including the inciting traumatic event, hospitalizations, multiple surgical interventions, postsurgical therapies, and disfigurement caused by non-hair-bearing scalp. Thus, as part of the medical management for scalp replant patients, one must address the psychological factors surrounding the medical management. Over the past 25 years, the authors have experienced four cases of scalp replant failures, each posing an opportunity to examine the postoperative course of these patients. Symptoms ranging from mild anxiety to depressive symptoms have been observed in all of these patients. In fact, patient symptoms often satisfied the criteria for major depressive disorder or posttraumatic stress disorder. The authors recognize the importance of informing patients and their families of the immediate and potential long-term complications following an unsuccessful scalp replant attempt. The authors advise that all patients be provided immediate psychiatric evaluation and, if necessary, counseling and medication therapy, regardless of scalp replantation outcome. PMID- 15114117 TI - The lower trapezius musculocutaneous flap from pedicled to free flap: anatomical basis and clinical applications based on the dorsal scapular artery. AB - The pedicled lower trapezius musculocutaneous flap is a standard flap in head and neck reconstruction. A review of the literature showed that there is no uniform nomenclature for the branches of the subclavian artery and the vessels supplying the trapezius muscle and that the different opinions on the vessels supplying this flap lead to confusion and technical problems when this flap is harvested. This article attempts to clarify the anatomical nomenclature, to describe exactly how the flap is planned and harvested, and to discuss the clinical relevance of this flap as an island or free flap. The authors dissected both sides of the neck in 124 cadavers to examine the variations of the subclavian artery and its branches, the vessel diameter at different levels, the course of the pedicle, the arc of rotation, and the variation of the segmental intercostal branches to the lower part of the trapezius muscle. Clinically, the flap was used in five cases as an island skin and island muscle flap and once as a free flap. The anatomical findings and clinical applications proved that there is a constant and dependable blood supply through the dorsal scapular artery (synonym for the deep branch of the transverse cervical artery in the case of a common trunk with the superficial cervical artery) as the main vessel. Harvesting an island flap or a free flap is technically demanding but possible. Planning the skin island far distally permitted a very long pedicle and wide arc of rotation. The lower part of the trapezius muscle alone could be classified as a type V muscle according to Mathes and Nahai because of its potential use as a turnover flap supplied by segmental intercostal perforators. The lower trapezius flap is a thin and pliable musculocutaneous flap with a very long constant pedicle and minor donor-site morbidity, permitting safe flap elevation and the possibility of free-tissue transfer. PMID- 15114118 TI - Estimation of breast burn size. AB - It is the authors' opinion that the size of chest burns on large-breasted women can be significantly underestimated, especially if the methods of calculation rely on burn charts, such as the Lund and Browder burns chart. This latter chart is based on data derived from only three women and eight men. The surface area of the torsos of 60 volunteers (20 men, 20 small-breasted women, and 20 large breasted women) was measured using two well-established techniques. The torso surface area was divided into two parts: the anterior trunk and the posterior trunk (i.e., torso surface area = posterior trunk + anterior trunk). The anterior trunk was subdivided and the area above the costal margins defined as the pectoral region. These areas were measured separately for each individual. The volunteers' total body surface area was calculated using normograms, based on their weight and height. The area of each torso section was recorded as a percentage of the total body surface area and torso surface area. Whereas the torso surface area/total body surface area ratio did not vary significantly between the groups, the proportion of anterior to posterior trunk size did depend on the sex and on breast size. There was a direct correlation between the woman's bra cup size and the ratio of anterior-to-posterior trunk surface area. A simple chart was therefore derived that estimates the relative size of a woman's torso surface area once her bra cup size is known. Such a chart can be used to improve accuracy in adult female chest burn estimation, when used in conjunction with a burns chart. Breast burns in larger breasted women are underestimated when calculated using current burn charts. We recommend that a correction be made when estimating chest burns in women to account for the increased surface area of the breasts. A chart, such as the one we have developed, could be used in conjunction with a burn chart (e.g., Lund and Browder) to make this correction. PMID- 15114119 TI - Investigation into the possible cause of subjective decreased sensory perception in the nipple-areola complex of women with macromastia. AB - Patients with macromastia often comment on a lack of sensation in their nipple areola complex. A study was designed to investigate the cause of this decreased sensation. Two hypotheses were proposed. First, the decreased sensation could result from neuropraxia of the sensory nerve fibers secondary to traction caused by the heavy breast parenchyma. The second hypothesis proposed that tissue expansion of the nipple and areola by the voluminous breast parenchyma caused a decrease in nerve fibers per surface area and hence decreased sensory perception. Sixty-one patients were assessed in the study. All patients underwent surgery in which histological biopsy of either the areola alone (31 reduction mammaplasty patients) or the nipple and areola (30 mastectomy patients) was possible. Before surgery, each nipple-areola complex was tested with Weinstein Enhanced Sensory Test monofilaments as a quantitative test of tactile sensation. Breast cup size, ptosis, and weight of tissue excised were recorded to allow general assessment of the breast size. The nipple and areola biopsy specimens were assessed using immunohistochemistry (S-100 polyclonal antibody, Dako Z311) to measure nerve fiber count per unit area. Statistical analysis was undertaken to find any association among sensitivity, breast cup size, ptosis, weight of tissue resected, and nerve fiber density in the nipple and areola biopsy specimens. Sensitivity at the areola decreased with increasing breast cup size (r = 0.47, p < 0.001) and ptosis (r = 0.42, p = 0.002 for increasing distance between inframammary crease and nipple; r = 0.49, p < 0.001 for increasing manubrium to nipple distance). There was a weak correlation between nerve fiber density at the areola and breast cup size (r = -0.22, p = 0.1). Sensitivity at the nipple was higher than at the areola. Nerve fiber density count at the nipple was higher than at the areola, but there was no statistically significant correlation between nipple sensitivity and breast cup size, ptosis, or weight of tissue resected. The results suggest that the areola and nipple are different in their neuroanatomy. The areola is a thin, pliable structure that is predisposed to stretch as the breast enlarges and therefore experience a decrease in nerve fiber density. The nipple is a compact structure that is less likely to stretch with breast enlargement. In the nipple, neither sensory perception nor nerve fiber density varied with size or breast ptosis. The perceived lack of sensation in the nipple-areola complex is multifactorial. This study shows that neither traction injury to the sensory nerves nor decreased nerve density alone can explain the subjective numbness reported by patients with macromastia. Psychological factors, such as dissatisfaction with body form or interpretation of lack of sensation in the areola as also affecting the nipple, may influence the patient's assessment of the nipple-areola sensitivity. PMID- 15114120 TI - Breast-conserving therapy in patients with a relatively large (T2 or T3) breast cancer: long-term local control and cosmetic outcome of a feasibility study. AB - Breast-conserving therapy is widely accepted as an appropriate method of primary treatment of T1 and T2 breast cancers that measure up to 5 cm. For safe and cosmetically acceptable breast-conserving therapy in patients with larger breast cancers, the tumor volume has to be reduced preoperatively, and lost tissue volume should be replaced after wide local excision. In 1993, the authors' group reported encouraging short-term results of a combination of preoperative radiotherapy, breast-conserving surgery, and immediate tissue replacement by myocutaneous (myosubcutaneous) latissimus dorsi flap transplantation in patients with relatively large T2 and T3 breast cancers. To evaluate the long-term oncologic local control and cosmetic outcome of this treatment modality, the authors studied the results obtained in 20 patients after a minimum follow-up of 5 years. The local control rate was 0.95, as locoregional recurrence was observed in one patient who refused adjuvant chemotherapy. Both the observed 5-year survival (0.75) and the actuarial 10-year survival (0.60) in the authors' series equaled that of more radical surgical therapy. The cosmetic outcome compared with that obtained by conventional breast-conserving therapy modalities for small breast cancers. In general, patient assessment of cosmetic outcome (2.8 of 3) was higher than the assessment of a professional panel (6.3 of 10). Of six criteria providing a detailed description of the cosmesis of the reconstructed breast and donor area, the symmetry and shape of the reconstructed breast were felt to be most important by the patients and professionals alike. The authors conclude that breast-conserving therapy combining preoperative irradiation and immediate myocutaneous (myosubcutaneous) latissimus dorsi flap reconstruction is an oncologically safe and cosmetically rewarding but logistically straining modality of treatment of relatively large T2 and T3 breast cancers. PMID- 15114121 TI - Delayed-immediate breast reconstruction. AB - In patients with early-stage breast cancer who are scheduled to undergo mastectomy and desire breast reconstruction, the optimal timing of reconstruction depends on whether postmastectomy radiation therapy will be needed. Immediate reconstruction offers the best aesthetic outcomes if postmastectomy radiation therapy is not needed, but if postmastectomy radiation therapy is required, delayed reconstruction is preferable to avoid potential aesthetic and radiation delivery problems. Unfortunately, the need for postmastectomy radiation therapy cannot be reliably determined until review of the permanent tissue sections. The authors recently implemented a two-stage approach, delayed-immediate breast reconstruction, to optimize reconstruction in patients at risk for requiring postmastectomy radiation therapy when the need for postmastectomy radiation therapy is not known at the time of mastectomy. Stage 1 consists of skin-sparing mastectomy with insertion of a completely filled textured saline tissue expander. After review of permanent sections, patients who did not require post-mastectomy radiation therapy underwent immediate reconstruction (stage 2) and patients who required postmastectomy radiation therapy completed postmastectomy radiation therapy and then underwent standard delayed reconstruction. In this study, the feasibility and outcomes of this approach were reviewed. Fourteen patients were treated with delayed-immediate reconstruction between May of 2002 and June of 2003. Twelve patients had unilateral reconstruction and two patients had bilateral reconstruction, for a total of 16 treated breasts. All patients completed stage 1. Tissue expanders were inserted subpectorally in 15 breasts and subcutaneously in one breast. The mean intraoperative expander fill volume was 475 cc (range, 250 to 750 cc). Three patients required postmastectomy radiation therapy and underwent delayed reconstruction. Eleven patients did not require postmastectomy radiation therapy. Nine patients had 11 breast reconstructions (stage 2), six with free transverse rectus abdominis musculocutaneous (TRAM) flaps, one with a superior gluteal artery perforator flap, and four with a latissimus dorsi flap plus an implant. The median interval between stages was 13 days (range, 11 to 22 days). Two patients who did not require postmastectomy radiation therapy have not yet had stage 2 reconstruction, one because she wished to delay reconstruction and the other because she required additional tissue expansion before permanent implant placement. Six complications occurred. The stage 1 complications involved two cases of mastectomy skin necrosis in patients who required post-mastectomy radiation therapy; one patient required removal of the subcutaneously placed expander before postmastectomy radiation therapy and the other patient had a subpectorally placed expander that only required local wound care. The stage 2 complications were a recipient-site seroma in a patient with a latissimus dorsi flap, a recipient-site hematoma in the patient with the superior gluteal artery perforator flap, and two arterial thromboses in patients with TRAM flaps. Both TRAM flaps were salvaged. Delayed-immediate reconstruction is technically feasible and safe in patients with early-stage breast cancer who may require postmastectomy radiation therapy. With this approach, patients who do not require postmastectomy radiation therapy can achieve aesthetic outcomes essentially the same as those with immediate reconstruction, and patients who require postmastectomy radiation therapy can avoid the aesthetic and radiation delivery problems that can occur after an immediate breast reconstruction. PMID- 15114122 TI - Which is the best position for the remote injection dome using the adjustable expander/prosthesis in breast reconstruction? A comparative study. AB - Breast reconstruction using mammary implants is a routinely performed surgical procedure that gives good aesthetic results with a relatively simple operation for the patients. When an adjustable expander/prosthesis with remote dome is used for reconstruction, the device is filled through an injection dome connected to the implant through a filling tube. The injection dome is usually inserted into a subcutaneous pocket, either in the axillary area or, most frequently, in the lower lateral thoracic area. Sometimes, this location is not well tolerated by the patient because of pain or discomfort in the breast-thoracic area and can give problems related to the distance, which causes kinking of the filling tube. To avoid this inconvenience and because of frequent patient complaints, the authors decided 3 years ago to place the injection dome in a parasternal position and compare this location with the previously used lower lateral thoracic location. Two hundred sixty patients were divided into two groups (130 patients in each group) and evaluated. All patients underwent mammary reconstruction in the authors' department using Becker adjustable implants. In all patients, the injection microdome was used. In group A, the injection microdome was positioned in the lower lateral thoracic area; in group B, the injection microdome was positioned in a parasternal area. Both groups were compared, considering different features such as pain, discomfort, ease of injection, pain during puncture, aesthetic appearance, risk of kinking, and risk of upside-down rotation of the dome. Average follow-up was 1.6 years. Statistical analysis was performed using Pearson's chi-square test regarding the differences in frequency of two features-aesthetic appearance and pain during puncture-between the two groups. The comparisons regarding both aesthetic appearance and pain during puncture did show a significant difference between the two groups, with a value of p < 0.05 in both cases. In the present study, the results showed how the patients had less pain during puncture and a better aesthetic appearance when the microdome was located in the parasternal position instead of the lower lateral thoracic area. Advantages and disadvantages of the locations used are discussed in this article. PMID- 15114123 TI - The infected or exposed breast implant: management and treatment strategies. AB - Among the potential complications associated with the use of breast implants are the risks of periprosthetic infection and device extrusion. There is little published information about the effective management of these situations. Conservative recommendations include antibiotic therapy and removal of the implant until resolution of the infection or until the wound has healed. A retrospective review identified patients with periprosthetic infection or threatened or actual device exposure treated by the senior author. Twenty-four patients encompassing 26 affected prostheses were available for review and were classified into seven groups based on initial presentation as follows: group 1, mild infection (n = 8); group 2, severe infection (n = 4); group 3, threatened exposure without infection (n = 3); group 4, threatened exposure with mild infection (n = 3); group 5, threatened exposure with severe infection (n = 1); group 6, actual exposure without clinical infection (n = 5); and group 7, actual exposure with infection (n = 2). To salvage the prosthesis in these patients, various treatment strategies were utilized. All patients with a suspected infection or device exposure were started immediately on appropriate antibiotic therapy (oral antibiotics for mild infections and parenteral antibiotics for severe infections). Salvage methods included one or more of the following: antibiotic therapy, debridement, curettage, pulse lavage, capsulectomy, device exchange, primary closure, and/or flap coverage. Twenty (76.9 percent) of 26 threatened implants with infection or threatened or actual prosthesis exposure were salvaged after aggressive intervention. The presence of severe infection adversely affected the salvage rate in this series. A statistically significant difference exists among those patients without infection or with mild infection only (groups 1, 3, 4, and 6); successful salvage was achieved in 18 (94.7 percent) of 19 patients, whereas only two of seven of those implants with severe infection (groups 2, 5, and 7) were salvaged (p = 0.0017). Ten (90.9 percent) of 11 devices with threatened or actual exposure, not complicated by severe infection (groups 3, 4, and 6), were salvaged. Several treatment strategies were developed for periprosthetic infection and for threatened or actual implant exposure. Patients with infection were placed on oral or intravenous antibiotics; those who responded completely required no further treatment. For persistent mild infection or threatened or actual exposure, operative intervention was required, including some or all of the following steps: implant removal, pocket curettage, partial or total capsulectomy, debridement, site change, placement of a new implant, and/or flap coverage; the menu of options varied with the precise circumstances. No immediate salvage was attempted in five cases, due to either severe infection, nonresponding infection with gross purulence, marginal tissues, or lack of options for healthy tissue coverage. Based on the authors' experience, salvage attempts for periprosthetic infection and prosthesis exposure may be successful, except in cases of overwhelming infection or deficient soft-tissue coverage. Although an attempt at implant salvage may be offered to a patient, device removal and delayed reinsertion will always remain a more conservative and predictable option. PMID- 15114124 TI - TRAM flap breast reconstruction: tumescent technique reduces blood loss and transfusion requirement. AB - The tumescent technique has been shown to be efficacious in reducing both operative and postoperative bleeding without significant deleterious side effects in suction lipectomy. In this study, the effects of the tumescent technique on postoperative complications in transverse rectus abdominis myocutaneous (TRAM) flap breast reconstruction are investigated. All women who underwent a TRAM flap breast reconstruction by the senior author (J.B.) at the Emory Clinic during the years 1990 to 1996 were pooled (n = 386). Any woman who had a preincision infiltration of 0.25% epinephrine-containing saline solution (>200 cc) around the donor site was included in the tumescent group (n = 59). Medical records were reviewed, and rates of partial flap loss, fat necrosis (> or =10 percent flap volume), flap full-thickness skin loss, donor-site complication (skin loss, hernia, or infection), and blood transfusion were determined. Group rates were compared. The infiltrated group had a significantly lower transfusion rate as compared with the control group (0.34 units versus 1.32 units, p < 0.001). The rates of partial flap loss and fat necrosis were less in the tumescent group, but not significantly (0 percent versus 4 percent, p = 0.232; and 1.7 percent versus 10.4 percent, p = 0.058). There were no significant differences in the incidence of full-thickness skin loss or donor-site complications. Donor-site infiltration before incision with a 0.25% epinephrine-containing saline solution significantly reduced the transfusion requirement in TRAM flap breast reconstruction patients without adversely affecting either breast mound or abdominal donor-site complication rates. PMID- 15114125 TI - Comparison of the deep inferior epigastric perforator flap and free transverse rectus abdominis myocutaneous flap in postmastectomy reconstruction: a cost effectiveness analysis. AB - This study compared the deep inferior epigastric perforator (DIEP) flap and the free transverse rectus abdominis myocutaneous (TRAM) flap in postmastectomy reconstruction using a cost-effectiveness analysis. A decision analytic model was used. Medical costs associated with the two techniques were estimated from the Ontario Ministry of Health Schedule of Benefits for 2002. Hospital costs were obtained from St. Joseph's Healthcare, a university teaching hospital in Hamilton, Ontario, Canada. The utilities of clinically important health states related to breast reconstruction were obtained from 32 "experts" across Canada and converted into quality-adjusted life years. The probabilities of these various clinically important health states being associated with the DIEP and free TRAM flaps were obtained after a thorough review of the literature. The DIEP flap was more costly than the free TRAM flap ($7026.47 versus $6508.29), but it provided more quality-adjusted life years than the free TRAM flap (28.88 years versus 28.53 years). The baseline incremental cost-utility ratio was $1464.30 per quality-adjusted life year, favoring adoption of the DIEP flap. Sensitivity analyses were performed by assuming that the probabilities of occurrence of hernia, abdominal bulging, total flap loss, operating room time, and hospital stay were identical with the DIEP and free TRAM techniques. By assuming that the probability of postoperative hernia for the DIEP flap increased from 0.008 to 0.054 (same as for TRAM flap), the incremental cost-utility ratio changed to $1435.00 per quality-adjusted life year. A sensitivity analysis was performed for the complication of hernia because the DIEP flap allegedly diminishes this complication. Increasing the probability of abdominal bulge from 0.041 to 0.103 for the DIEP flap changed the ratio to $2731.78 per quality-adjusted life year. When the probability of total flap failure was increased from 0.014 to 0.016, the ratio changed to $1384.01 per quality-adjusted life year. When the time in the operating room was assumed to be the same for both flaps, the ratio changed to $4026.57 per quality-adjusted life year. If the hospital stay was assumed to be the same for both flaps, the ratio changed to $1944.30 per quality-adjusted life year. On the basis of the baseline calculation and sensitivity analyses, the DIEP flap remained a cost-effective procedure. Thus, adoption of this new technique for postmastectomy reconstruction is warranted in the Canadian health care system. PMID- 15114126 TI - Evaluation of abdominal wall strength after TRAM flap surgery. AB - Evaluation of abdominal wall function after transverse rectus abdominis musculocutaneous (TRAM) flap surgery has been mostly subjective. The purpose of this study was to measure abdominal wall strength objectively and to compare the results with the patient's performance of daily activities. Abdominal wall strength was objectively measured with the B200 IsoStation machine preoperatively and 1 year after TRAM flap breast reconstruction. These data were compared with the results of a questionnaire evaluating the patient's performance of daily activities. The results of this testing in 21 patients demonstrated the following: (1) a decrease in abdominal wall strength after bilateral pedicled TRAM flap surgery, which was significant in trunk flexion (34.2 +/- 16.9 ft-lbs to 20.6 +/- 15.2 ft-lbs); (2) compensation by other truncal musculature, with an increase in the strength of trunk rotation (18.8 +/- 13.5 ft-lbs to 28.6 +/- 17.7 ft-lbs) seen after unilateral pedicled TRAM flap surgery; (3) minimal interference with the patient's daily activities; and (4) no effect of mesh on strength. A relationship was demonstrated between the degree of loss of strength in trunk flexion and the patient's difficulty in performing certain activities. The patients who had the greatest loss in trunk flexion following the use of both pedicles also had the most difficulty in performing some daily activities. The patients were satisfied with their reconstructive procedure, with an average score of 8.3 (on a scale of 1 to 10), and reported an improvement in the appearance of their abdomen, with an average increase of 5.1 to 6.8 (on a scale of 1 to 10). PMID- 15114128 TI - A surgical approach for earlobe keloid: keloid fillet flap. AB - Earlobe keloid can form after cosmetic ear piercing, trauma, or burns, and it poses several difficulties in treatment and distinctive cosmetic implications. Treatment methods for earlobe keloids include both surgical and nonsurgical methods. After excision of the earlobe keloid, healing by secondary intention, primary suture, skin graft, or local flap has revealed some disadvantages. The authors approached this problem with a new excision and covering method. The surgery was performed under local anesthesia. Skin over the keloid was dissected from the keloid mass as a flap, which they termed a "keloid fillet flap," and the keloid mass was completely removed. Subcutaneous sutures were not used, and the keloid fillet flaps were closed with 6-0 nylon sutures after trimming. Other intraoperative or postoperative preventive procedures, such as steroid injection, pressure device, or irradiation, were not applied primarily. In the period from May of 1999 to October of 2000, nine earlobe keloids in eight patients were treated with this protocol. One patient had bilateral keloids. Of the eight patients, there were six women and two men, ranging in age from 21 to 61 years (mean age, 28.5 years). The causes of keloids were ear piercing in six cases and trauma in three cases. The largest lesion was 3 cm in its greatest dimension, and the smallest was 1.5 cm (mean, 2.3 cm). All flaps survived completely. There were four cases of recurrence. Seven cases, including two recurrences, showed good results. The authors believe the recurrence of earlobe keloid was closely related to the method for coverage of the defect after its surgical excision, and the "5 As and one B" (Asepsis, Atraumatic technique, Absence of raw surface, Avoidance of tension, Accurate approximation of wound margin, and complete Bleeding control) are important factors in reducing the recurrence rate of earlobe keloids in surgical excision. The authors' protocol is very effective in closing the defect after surgical excision of earlobe keloids and offers many advantages over other surgical approaches. The recurrence rate of earlobe keloid may be lower than in their results if other intraoperative and postoperative treatment procedures are combined with their protocol. PMID- 15114129 TI - TGF-beta1, FGF-2, and receptor mRNA expression in suture mesenchyme and dura versus underlying brain in fusing and nonfusing mouse cranial sutures. AB - Recent studies have supported a functional role for the transforming growth factor beta-1 (TGF-beta1) and fibro-blast growth factor 2 (FGF-2) signaling cascades in the process of mouse cranial suture fusion. TGF-beta1 and FGF-2 protein expression have been shown to be elevated in the fusing posterior frontal suture versus the nonfusing sagittal suture. The authors evaluated simultaneous mRNA expression of TGF-beta1 and its R1 receptor and FGF-2 and its R2 receptor during mouse cranial suture fusion. They evaluated the suture mesenchyme-dura complex separately from the underlying brain to determine whether there is tissue specific biologic activity (i.e., brain versus suture mesenchyme-dura) for each cytokine and receptor. Data were collected from 150 male CD-1 mice studied over five time periods from postnatal days 22 to 45. They utilized reverse transcriptase polymerase chain reaction as a means to detect TGF-beta1, TGF-beta receptor 1 (TGF-betaR1), FGF-2, and FGF receptor 2 (FGFR2) mRNA expression in mouse cranial tissues, beginning with the period of initiation of posterior frontal cranial suture fusion (postnatal day 22) and extending through completion of posterior frontal suture fusion (postnatal day 45). Expression of FGF-2 was significantly greater in posterior frontal suture mesenchyme and dura compared with sagittal suture mesenchyme and dura during the period of initiation of posterior frontal suture fusion, localizing this cytokine's expression to posterior frontal suture mesenchyme and dura during the process of cranial suture fusion. TGF-beta1 and FGFR2 mRNA expression was found to be up-regulated in posterior frontal suture mesenchyme and dura relative to the underlying brain tissue throughout the study period, whereas TGF-betaR1 and FGF-2 mRNA expression was significantly elevated relative to the underlying brain only at time points corresponding to the initiation of posterior frontal suture fusion (between postnatal days 22 and 31). These results indicate that there is tissue-specific mRNA expression of TGF-beta1, FGF-2, and their receptors between suture mesenchyme and dura and the underlying brain, which correlates with the period of posterior frontal suture fusion in the mouse model. Differences in gene expression between suture mesenchyme and dura relative to the underlying brain may be an important regulator of cranial suture biology. Understanding these differences may eventually help to identify possible targets and time windows by which to most effectively modulate cranial suture fusion. PMID- 15114130 TI - Mechanisms of murine cranial suture patency mediated by a dominant negative transforming growth factor-beta receptor adenovirus. AB - Using a physiologic model of mouse cranial suture fusion, the authors' laboratory has previously demonstrated that transforming growth factor (TGF)-betas appear to be more abundantly expressed in the suture complex of the fusing posterior frontal compared with the patent sagittal suture. Furthermore, the authors have shown that by blocking TGF-beta signaling with a replication-deficient adenovirus encoding a defective, dominant negative type II TGF-beta receptor (AdDN TbetaRII), posterior frontal suture fusion was inhibited. In this study, the authors attempt to further elucidate the role of TGF-beta in cranial suture fusion by investigating possible mechanisms of AdDN-TbetaRII-mediated cranial suture patency using both an established organ culture model and a novel in vitro co-culture system that recapitulates the in vivo anatomic dura mater/cranial suture relationship. In this article, the authors demonstrate that blocking TGF beta signaling with the AdDN-TbetaRII construct led to inhibition of cellular proliferation in the suture mesenchyme and subjacent dura mater during the early period of predicted posterior frontal suture fusion. Interestingly, co-culture experiments revealed that transfecting osteoblasts with AdDN-TbetaRII led to alterations in the gene expression levels of two important bone-related molecules (Msx2 and osteopontin). Inhibiting TGF-beta signaling prevented time-dependent suppression of Msx2 and prevented induction of osteopontin, thereby retarding osteoblast differentiation. Furthermore, the authors demonstrated that the AdDN TbetaRII construct was capable of blocking TGF-beta -mediated up-regulation of collagen IalphaI, an extracellular matrix molecule important for bone formation. Collectively, these data strongly suggest that AdDN-TbetaRII maintains posterior frontal patency, in part by altering early events in de novo bone formation, including cellular proliferation and early extracellular matrix production. PMID- 15114131 TI - Effect of L-arginine on leukocyte adhesion in ischemia-reperfusion injury. AB - Nitric oxide has been reported to be beneficial in preserving muscle viability following ischemia-reperfusion injury. The purpose of this study was to evaluate the influence of nitric oxide via L-arginine on leukocyte adhesion following ischemia-reperfusion injury. Intravital videomicroscopy of rat gracilis muscle was used to quantify changes in leukocyte adherence. The gracilis muscle was raised on its vascular pedicle in 48 male Wistar rats. The animals were assigned to one of five groups: (1) nonischemic control; (2) ischemia-reperfusion; (3) ischemia-reperfusion and L-arginine; (4) ischemia-reperfusion and Nomega-nitro-L arginine methyl ester (L-NAME); and (5) ischemia-reperfusion, L-NAME, and L arginine. All groups that included ischemia-reperfusion were subjected to 4 hours of global ischemia followed by 2 hours of reperfusion. L-Arginine (10 mg/kg) and L-NAME (10 mg/kg) were infused into the contralateral femoral vein beginning 5 minutes before reperfusion, for a total of 30 minutes. The number of adherent leukocytes was counted at baseline and at 5, 15, 30, 60, and 120 minutes after reperfusion (reported as mean change from baseline, +/- SEM). Groups were compared by repeated-measures analysis of variance (five groups, five times). P < or =0.05 was accepted as significant. L-Arginine significantly reduced leukocyte adherence to venular endothelium during reperfusion when compared with the ischemia-reperfusion group (1.39 +/- 0.92 versus 12.78 +/- 1.43 at 2 hours, p < 0.05). Administration of L-NAME with L-arginine showed no significant difference in adherent leukocytes when compared with the ischemia-reperfusion group (10.28 +/- 2.03 at 2 hours). The nitric oxide substrate L-arginine appears to reduce the deleterious neutrophil-endothelial adhesion associated with ischemia-reperfusion injury. L-NAME (nitric oxide synthesis inhibitor) given concomitantly with L arginine reversed the beneficial effect of L-arginine alone, indicating that L arginine may be acting via a nitric oxide synthase pathway. These results suggest an important role for nitric oxide in decreasing the neutrophil-endothelial interaction associated with ischemia-reperfusion injury. PMID- 15114132 TI - Tendon healing in vitro: activation of NIK, IKKalpha, IKKbeta, and NF- kappaB genes in signal pathway and proliferation of tenocytes. AB - Initiation of DNA transcription and proliferation of tendon cells are critical to tendon healing and require pivotal signals to the nucleus. Exploring intracellular signaling pathways pertinent to the healing process may reveal new approaches to accelerating the healing rate of the tendon. The authors investigated expression of NIK, IKKalpha, IKKbeta, and NF- kappaB genes in the signal pathway and tenocyte proliferation in an in vitro model in which cultured tenocytes were exposed to basic fibroblast growth factor (bFGF). Tenocytes were obtained from explant culture of rabbit intrasynovial tendons and were treated with bFGF at concentrations of 0, 2, or 10 ng/ml. Levels of expression of a series of genes for key factors along the signaling route--nuclear factor (NF) kappaB-inducing kinase, inhibitor of kappa B kinase alpha and beta, and the NF kappaB--were examined by quantitative analysis of products of reverse transcription and multiplex polymerase chain reactions. Proliferation of the cells was assessed with evaluation of growth curves and immunochemical labeling of the DNA of the cells. Expression levels of NIK, IKKalpha, IKKbeta, and NF kappaB genes were significantly increased by bFGF at concentrations of 2 and 10 ng/ml. Western blot confirmed the increase of NF-kappaB in the tenocytes. The proliferation rate of the cells was significantly promoted by bFGF. Expression of these genes increased proportionately to the amounts of bFGF stimulating the cells and was correlated with increases in the proliferation rate. This study showed that expression of a series of genes along the NF-kappaB pathway was remarkably promoted by bFGF. The effects were proportionate to in vitro cell proliferation rate. Results of the study suggest that activation of a series of genes along the NF-kappaB pathway may play a pivotal role in initiating cell proliferation during the healing process of intrasynovial tendons. As activation of genes in signal transduction pathways is a new field in the biology of growth factor action with tremendous potential in promoting tissue repairs, manipulation of expression of a series of genes along the NF-kappaB pathway can be a new target of enhancing tendon healing through molecular mechanisms. PMID- 15114133 TI - Force deficits in skeletal muscle after delayed reinnervation. AB - Using a rat hindlimb model, the authors tested the hypothesis that, in muscles reinnervated after long-term denervation, atrophy-dependent and atrophy independent mechanisms operate independently to produce force deficits. In adult rats, gastrocnemius muscles were subjected to denervation via tibial nerve transection. Reconstruction of the nerve lesion was delayed for periods ranging from 2 weeks to 1 year. After a minimum recovery period of 6 months after nerve repair, muscle mass and maximum isometric tetanic force were measured and specific force was calculated for each muscle (n = 40 muscles from 23 animals). After recovery, observed deficits in muscle mass and maximum tetanic force were directly proportional to the denervation interval. On the other hand, the deficit in specific force was not proportional to the denervation interval; all groups in which the nerve reconstruction was delayed for a month or longer demonstrated a deficit of 30 percent to 50 percent. These data support our hypothesis that, after prolonged denervation followed by reinnervation, the magnitude of the deficit in whole muscle force does not parallel the deficit in specific force. These data support the idea that mechanisms governing muscle atrophy are independent of those resulting in specific force deficits. PMID- 15114134 TI - The role of allogenic fibroblasts in an acute wound healing model. AB - Skin is the first tissue-engineered organ to have been successfully developed in the laboratory, and it has been clinically available for use as epidermal sheets for some time. As refinements in this field of tissue engineering continue, several key issues give cause for concern. One issue is the need to form a more complete dermal analogue before grafting. To this end, fibroblasts may be used in vitro to deposit extracellular matrix components within a basic scaffold, laying down those molecules not endogenous to the material and thereby improving the quality of the skin replacement. Many studies have shown the benefits of in vitro seeding with fibroblasts, but there has been some debate regarding the longevity of such cells after allotransplantation into an immunocompetent host. In this study, the authors set out to determine the longevity of transplanted cells in an immunocompetent porcine model. A total of 24 wounds were made on four female animals, 12 of which were covered with acellular hyaluronic acid dermal matrices, and the remainder of which were covered with matrices seeded with allogenic (male) fibroblasts. After a week in vivo, the wounds were grafted with either split-thickness skin grafts or cultured epithelial autograft. Biopsy specimens were obtained from wounds at varying time intervals and assessed using genetic analysis to determine the survival of allotransplanted cells. No cells were detectable by polymerase chain reaction analysis (sensitivity, 1:100,000) after 7 days in vivo. Subsequent histologic examination demonstrated little difference in wound morphology. The authors conclude that allogenic fibroblasts do not survive transplantation in a porcine wound model. PMID- 15114135 TI - Transient total facial palsy after bilateral sagittal split ramus osteotomy. PMID- 15114136 TI - Bilateral high upper limb replantation in a child. AB - Bilateral high amputation of upper limbs in a child is a very unusual injury. In the present case, although the amputation was high and significant avulsion was present, the age of the child (6 years) made the case both challenging and encouraging--challenging because of the anticipated systemic effects of reperfusion, and encouraging because the long-term prognosis is always more encouraging in a child. PMID- 15114138 TI - A nonexcision plication method for treatment of bilateral congenital boutonniere deformity: a preoperative biomechanical calculation. PMID- 15114139 TI - Salvage of the massively traumatized lower extremity with sequential free flaps. PMID- 15114140 TI - Further clinical experience with orbicularis oculi myocutaneous flaps in the temporal area. PMID- 15114141 TI - The reverse digital artery island flap: an update. PMID- 15114142 TI - Unilateral and bilateral metacarpal hand injuries: classification and treatment guidelines. PMID- 15114143 TI - Analysis of outpatient surgery center safety using an internet-based quality improvement and peer review program. AB - Assessing the quality of care delivered in office-based outpatient surgery centers is difficult because formerly there was no central data collection system. The American Association for Accreditation of Ambulatory Surgery Facilities (AAAASF), in its ongoing effort to assess and improve patient care, has developed an Internet-based quality improvement and peer review program to analyze outcomes for surgery centers it accredits. Reporting is mandatory for all surgeons operating in AAAASF-accredited facilities. Each surgeon must report all unanticipated sequelae and at least six random cases reviewed by an accepted peer review group biannually. A total of 411,670 procedures were analyzed during a 2 year period (from 2001 to 2002). There were 2597 sequelae reported during this period. The most common sequela was hematoma formation following breast augmentation. Infection occurred in 388 cases. Deep vein thrombosis, pulmonary embolism, and intraoperative cardiac arrhythmias were found to occur in a frequency consistent with previous reports. Significant complications (hematoma, hypertensive episode, wound infection, sepsis, and hypotension) were infrequent. A total of 1378 significant sequelae were reported for 411,670 procedures. This calculates to one unanticipated sequela in 299 procedures (an incidence of 0.33 percent). Seven deaths were reported. A death occurred in one in 58,810 procedures (0.0017 percent). The overall risk of death was comparable whether the procedure was performed in an AAAASF-accredited office surgery facility or a hospital surgery facility. This study documents an excellent safety record for surgical procedures performed in accredited office surgery facilities by board certified surgeons. PMID- 15114144 TI - Liposuction of the legs and ankles: a review of the literature. PMID- 15114145 TI - The carrot or the carrot seed? PMID- 15114146 TI - Using intense pulsed light for cosmetic purposes: our experience. AB - The authors' experience using intense pulsed light for skin rejuvenation is summarized and analyzed with regard to its efficacy, safety, and complications. Rejuvenation using intense pulsed light was performed on 59 patients over a 6 month period (January of 2002 to July of 2002); these patients served as the study group. The areas treated were the face, neck, chest, hands, and legs. The parameters used during the procedure, patient satisfaction, and complications are described. Ninety-five percent of the patients included in the study had one or two sessions. Good to very good results were reported by 93.1 percent. Most patients had minor side effects and only three patients (5 percent) experienced complications (hyperpigmentation or scars). Intense pulsed light is an effective and safe method for skin rejuvenation. Its efficacy is mainly manifested by eliminating senile pigmentation and telangiectasias and a achieving a younger and fresher appearance of the skin. Although in the literature intense pulsed light skin rejuvenation is mainly reported for the face, the authors have obtained good results by using it for other areas of the body. The majority of the patients were satisfied. The authors conclude that intense pulsed light skin rejuvenation is a safe and effective method for facial and nonfacial rejuvenation. PMID- 15114147 TI - Effect of low-level laser therapy on abdominal adipocytes before lipoplasty procedures. AB - Low-level laser therapy is a new subspecialty for the medical application of lasers that provides therapeutic rather than surgical outcomes for many medical indications. Recently, low-level laser therapy was reported to "liquefy" or release stored fat in adipocytes by the opening of specialized yet not identified cell membrane-associated pores after a brief treatment. Currently, low-level laser therapy is a U.S. Food and Drug Administration-approved technology for improving pain alleviation. To explore these data further, a series of in vitro studies on human preadipocytes and institutional animal care and use committee approved protocols in a porcine Yucatan model and an institutional review board approved clinical study were performed. Using a 635-nm low-level laser of 1.0 J/cm supplied to the authors by the vendor, these studies were designed to determine whether alteration in adipocyte structure or function was modulated after low-level laser therapy. Cultured human preadipocytes after 60 minutes of laser therapy did not change appearance compared with nonirradiated control cells. In the porcine model, low-level laser therapy (30 minutes) was compared with traditional lipoplasty (suction-assisted lipoplasty) and ultrasound-assisted lipoplasty. From histologic and scanning electron microscopic evaluations of the lipoaspirates, no differences were observed between low-level laser therapy derived and suction-assisted lipoplasty-derived specimens. Using exposure times of 0, 15, 30, and 60 minutes in the presence or absence of superwet wetting solution and in the absence of lipoplasty, total energy values of 0.9 mW were delivered to tissue samples at three increasing depths from each experimental site. No histologic tissue changes or specifically in adipocyte structure were observed at any depth with the longest low-level laser therapy (60 minutes with superwet fluid). Three subjects undergoing large-volume lipoplasty were exposed to superwet wetting fluid infiltration 14 minutes before and 12 minutes after, according to vendor instructions. Tissue samples from infiltrated areas were collected before suction-assisted lipoplasty and lipoaspirates from suction assisted lipoplasty. No consistent observations of adipocyte disruptions were observed in the histologic or scanning electron microscopy photographs. These data do not support the belief that low-level laser therapy treatment before lipoplasty procedures disrupts tissue adipocyte structure. PMID- 15114149 TI - The outcome of abdominoplasty performed under conscious sedation: six-year experience in 153 consecutive cases. AB - The use of conscious sedation is rapidly gaining acceptance and popularity in plastic surgery. At the present time, many procedures are performed using intravenous sedation and local anesthesia. The purpose of this article was to examine the safety and outcome of full abdominoplasties performed under conscious sedation at the authors' institution. Over a 6-year period from 1997 to 2002, 266 abdominoplasties were performed by the two senior authors. One hundred thirteen of these (42 percent) were performed under a general or regional anesthetic because a concurrent procedure was performed that precluded the use of conscious sedation (64 hysterectomies, 18 hernia repairs, six urogynecologic procedures, 10 breast reductions, and one laparoscopic cholecystectomy) or because of patient and surgeon preference (14 cases). One hundred fifty-three abdominoplasties (58 percent) were performed under conscious sedation using intravenous midazolam and fentanyl along with a local anesthetic. No patients had an unplanned conversion to deep sedation or general anesthesia. Eighty percent of these cases were performed with a concurrent procedure (80 liposuctions, 19 breast augmentations, 20 mastopexies, three capsulotomies, and 13 varied facial aesthetic procedures). In addition, 12 patients had concurrent hernia repairs (five ventral and seven umbilical) under conscious sedation. Mean follow-up was 10 months (range, 1 to 56 months). There were no intraoperative complications and no major postoperative complications. The minor complication rate was 11.1 percent (10 seromas requiring needle aspiration in the office, three superficial wound infections, two cases of marginal skin necrosis, one stitch abscess, and one pseudobursa requiring reexcision). Seven revisions were performed for suboptimal scars (5 percent). The results of this study demonstrate that abdominoplasties can be performed under conscious sedation in a safe and cost-effective manner for almost all patients. This type of procedure is well tolerated, has a low complication rate, and has high patient satisfaction. Increasing experience and small modifications in local anesthesia and surgical technique have strengthened the authors' conviction that conscious sedation is the preferred method of anesthesia for most patients undergoing abdominoplasty. PMID- 15114151 TI - Central wedge nymphectomy with a 90-degree Z-plasty for aesthetic reduction of the labia minora. AB - The labia minora or nymphae of the vulva are two cutaneous-mucosal refolds located between the labia majora, the internal aspect of which is separated by the interlabial cleft. The enlargement of the labia minora may be attributable to several factors, most commonly congenital. Although some women require surgical reduction for functional reasons, most seek reduction of their labia minora because of psychological concerns. The authors describe a modified plastic surgery procedure for functional and aesthetic reduction of the labia minora. PMID- 15114153 TI - Lifting of the upper lip: personal technique. AB - The authors illustrate a personal technique for lifting of the upper lip with augmentation of the lower lip. With this procedure, a shortening of the "prolabium," an increase of the vermilion, and a natural, nicer mouth are obtained, with the possibility of increasing the volume of the lower lip simultaneously. The operation is carried out as outpatient surgery using local anesthesia, with intravenous sedation if requested. Incisions are made bilaterally beginning at the alar fold of the nose; they then enter the nostrils and rise medially on the skin below the lower margin of the medial crura of the alar cartilage. In this way, the columella is safe, and there are no scars. Then, the two pieces of excess skin and a small, whole strip of orbicular muscle can be cut away, just under the nose. If the goal is to better extrude the vermilion, the skin as far as the Cupid's bow also has to be undermined; if the goal is to shorten the prolabium, a slightly wider amount of orbicular muscle can be removed. The muscle is suspended to the base of the nose with interrupted stitches (absorbable 4-0 suture), the subcutaneous tissue is sutured, and finally the skin is closed with a running suture. The removed muscle is a good graft for increasing the size of the lower lip. PMID- 15114155 TI - Herpes zoster as a rare complication of liposuction. AB - Liposuction typically has a low incidence of complications and is associated with significant cosmetic benefit. In this case, the patient developed a dermatomal rash with vesicles on an erythematous base consistent with herpes zoster 8 days after liposuction to the back and flanks. To the authors' knowledge, herpes zoster has not been previously reported as a complication of liposuction. Although the precise relationship of herpes zoster infection to the liposuction procedure is difficult to determine, mechanical irritation most likely reactivated the varicella zoster virus in the involved dermatomal distribution. The patient was treated with antiviral and analgesic medications and healed without any further complications. PMID- 15114156 TI - Frontalis muscle advancement: a dynamic structure for the treatment of severe congenital eyelid ptosis. AB - Forty-two consecutive patients have had severe eyelid ptosis corrected by intraorbital frontalis flap advancement as a motor unit to substitute for the function of the levator muscle. This technique has avoided the need for the linking structure necessary in the standard frontalis sling approach and has improved the direction of pull to more closely mimic that of a normal levator. This simple technique includes elevation of the innervated frontalis muscle flap and the creation of a pulley near the insertion of the orbital septum at the superior orbital rim, which redirects the lid movement along the surface of the globe rather than lifting it from the globe's surface toward the brow. This type of displacement is produced because the muscle is directed posteriorly by the pulley, so that it conforms to the plane of the levator aponeurosis all the way down to the tarsal plate. In addition, to improve the remaining function of the levator muscle (if any) and to facilitate voluntary positioning of the eyelid, the levator aponeurosis is shortened by plication. Symmetry is created by intervention on the contralateral eyelid to provide symmetrical supratarsal creases. PMID- 15114158 TI - Personal experience with ultrasound-assisted lipoplasty: a pilot study comparing ultrasound-assisted lipoplasty with traditional lipoplasty. PMID- 15114160 TI - Insanity from innovation. PMID- 15114159 TI - Is posttraumatic first web contracture avoidable? Prophylactic guidelines and treatment-oriented classification. PMID- 15114162 TI - Submucous cleft palate: an aid to diagnosis. PMID- 15114161 TI - Winston Churchill as a skin donor. PMID- 15114163 TI - How many rhomboids? "Try a square peg into a round hole!". PMID- 15114164 TI - Conformational technique of dressing application. PMID- 15114165 TI - Arnica and homeopathy. PMID- 15114166 TI - Fatal pulmonary embolism. PMID- 15114168 TI - Foreign-body retrieval using a rare earth magnet. PMID- 15114169 TI - Simple solution for pectus excavatum repair. PMID- 15114170 TI - Facial anatomy of the eyelids. PMID- 15114172 TI - Reversed-flow external jugular vein: an optional recipient vessel in microsurgical head and neck reconstruction. PMID- 15114173 TI - Beware venous backflow in head and neck cancer reconstruction. PMID- 15114174 TI - Correction of thin lips. PMID- 15114175 TI - Deflating a breast prosthesis before operation. PMID- 15114177 TI - A helpful idea in breast augmentation. PMID- 15114178 TI - Late complications after injections of hydrogel in the breast. PMID- 15114180 TI - Seat belt injury to female chest. PMID- 15114181 TI - Ossifying calcification of breast implant capsule. PMID- 15114182 TI - Thumb metacarpophalangeal joint constriction due to burn contracture and its treatment with miniplate arthrodesis. PMID- 15114183 TI - Two-in-one flaps for resurfacing defects of the upper third of the leg. PMID- 15114184 TI - Combination of bilateral perforator flaps and fasciocutaneous flaps for coverage of large, expansive sacral pressure ulcer. PMID- 15114185 TI - Paraffinoma of the labia. PMID- 15114186 TI - Loupe magnification reduces the incidence of incomplete excision of basal cell carcinoma. PMID- 15114187 TI - Solitary benign schwannoma of the external oblique muscle. PMID- 15114188 TI - An easy technique for manual meshing of split-thickness skin grafts. PMID- 15114189 TI - V-Y-S plasty for scalp defects. PMID- 15114191 TI - How to prevent complications during the application of nasal plaster (the spider maneuver). PMID- 15114192 TI - Stick it up: a temporary aid to improve unilateral brow ptosis. PMID- 15114193 TI - Some thoughts on augmentation/mastopexy and medical malpractice. PMID- 15114194 TI - Spectacle-induced facial lesions are not that uncommon. PMID- 15114195 TI - Ultrasound-assisted lipoplasty treatment for axillary bromidrosis: clinical experience of 375 cases. PMID- 15114197 TI - Preoperative corticosteroids for reactive airway? PMID- 15114198 TI - No magic bullets: the ephemeral nature of anesthetic-mediated neuroprotection. PMID- 15114199 TI - Corticosteroids and inhaled salbutamol in patients with reversible airway obstruction markedly decrease the incidence of bronchospasm after tracheal intubation. AB - BACKGROUND: In patients with bronchial hyperreactivity, airway instrumentation can evoke life-threatening bronchospasm. However, the best strategy for the prevention of bronchospasm has not been defined. Therefore, in a randomized, prospective, placebo-controlled study, the authors tested whether prophylaxis with either combined salbutamol-methylprednisolone or salbutamol alone (1) improves lung function and (2) prevents wheezing after intubation. METHODS: Thirty-one patients with partially reversible airway obstruction (airway resistance > 180%, forced expiratory volume in 1 s [FEV1] < 70% of predicted value, and FEV1 increase > 10% after two puffs of salbutamol), who were naive to anti-obstructive treatment, were randomized to receive daily for 5 days either 3 x 2 puffs (0.2 mg) of salbutamol alone (n = 16) or salbutamol combined with methylprednisolone (40 mg/day orally) (n = 15). Lung function was evaluated daily. Another 10 patients received two puffs of salbutamol 10 min before anesthesia. In all patients, wheezing was assessed before and 5 min after tracheal intubation. RESULTS: Within 1 day, both salbutamol and salbutamol methylprednisolone treatment significantly improved airway resistance (salbutamol, 4.3+/- 2.0 [SD] to 2.9+/-1.3 mmHg x s x l(-1); salbutamol methylprednisolone, 5.5+/-2.9 to 3.4+/-1.7 mmHg x s x l(-1)) and FEV1 (salbutamol, 1.79+/-0.49 to 2.12+/-0.61 l; salbutamol-methylprednisolone, 1.58+/ 0.66 to 2.04+/-1.05 l) to a steady state, with no difference between groups. However, regardless of whether single-dose salbutamol preinduction or prolonged salbutamol treatment was used, most patients (8 of 10 and 7 of 9) experienced wheezing after intubation. In contrast, only one patient receiving additional methylprednisolone experienced wheezing (P = 0.0058). CONCLUSIONS: : Pretreatment with either salbutamol alone or salbutamol combined with methylprednisolone significantly and similarly improves lung function within 1 day. However, only combined salbutamol-methylprednisolone pretreatment decreases the incidence of wheezing after tracheal intubation. Therefore, in patients with bronchial hyperreactivity, preoperative treatment with combined corticosteroids and salbutamol minimizes intubation-evoked bronchoconstriction much more effectively than the inhaled beta2-sympathomimetic salbutamol alone. PMID- 15114200 TI - Effects of a circulating-water garment and forced-air warming on body heat content and core temperature. AB - BACKGROUND: Forced-air warming is sometimes unable to maintain perioperative normothermia. Therefore, the authors compared heat transfer, regional heat distribution, and core rewarming of forced-air warming with a novel circulating water garment. METHODS: Nine volunteers were each evaluated on two randomly ordered study days. They were anesthetized and cooled to a core temperature near 34 degrees C. The volunteers were subsequently warmed for 2.5 h with either a circulating-water garment or a forced-air cover. Overall, heat balance was determined from the difference between cutaneous heat loss (thermal flux transducers) and metabolic heat production (oxygen consumption). Average arm and leg (peripheral) tissue temperatures were determined from 18 intramuscular needle thermocouples, 15 skin thermal flux transducers, and "deep" hand and foot thermometers. RESULTS: Heat production (approximately 60 kcal/h) and loss (approximately 45 kcal/h) were similar with each treatment before warming. The increases in heat transfer across anterior portions of the skin surface were similar with each warming system (approximately 65 kcal/h). Forced-air warming had no effect on posterior heat transfer, whereas circulating-water transferred 21+/-9 kcal/h through the posterior skin surface after a half hour of warming. Over 2.5 h, circulating water thus increased body heat content 56% more than forced air. Core temperatures thus increased faster than with circulating water than forced air, especially during the first hour, with the result that core temperature was 1.1 degrees +/- 0.7 degrees C greater after 2.5 h (P < 0.001). Peripheral tissue heat content increased twice as much as core heat content with each device, but the core-to-peripheral tissue temperature gradient remained positive throughout the study. CONCLUSIONS: The circulating-water system transferred more heat than forced air, with the difference resulting largely from posterior heating. Circulating water rewarmed patients 0.4 degrees C/h faster than forced air. A substantial peripheral-to-core tissue temperature gradient with each device indicated that peripheral tissues insulated the core, thus slowing heat transfer. PMID- 15114201 TI - Effects of subanesthetic ketamine on regional cerebral glucose metabolism in humans. AB - BACKGROUND: The authors have recently shown with positron emission tomography that subanesthetic doses of racemic ketamine increase cerebral blood flow but do not affect oxygen consumption significantly. In this study, the authors wanted to assess the effects of racemic ketamine on regional glucose metabolic rate (rGMR) in similar conditions to establish whether ketamine truly induces disturbed coupling between cerebral blood flow and metabolism. METHODS: 18F-labeled fluorodeoxyglucose was used as a positron emission tomography tracer to quantify rGMR on 12 brain regions of interest of nine healthy male volunteers at baseline and during a 300-ng/ml ketamine target concentration level. In addition, voxel based analysis was performed for the relative changes in rGMR using statistical parametric mapping. RESULTS: The mean +/- SD measured ketamine serum concentration was 326.4+/-86.3 ng/ml. The mean arterial pressure was slightly increased (maximally by 16.4%) during ketamine infusion (P < 0.001). Ketamine increased absolute rGMR significantly in most regions of interest studied. The greatest increases were detected in the thalamus (14.6+/-15.9%; P = 0.029) and in the frontal (13.6+/-13.1%; P = 0.011) and parietal cortices (13.1+/-11.2%; P = 0.007). Absolute rGMR was not decreased anywhere in the brain. The voxel-based analysis revealed relative rGMR increases in the frontal, temporal, and parietal cortices. CONCLUSIONS: Global increases in rGMR seem to parallel ketamine-induced increases in cerebral blood flow detected in the authors' earlier study. Therefore, ketamine-induced disturbance of coupling between cerebral blood flow and metabolism is highly unlikely. The previously observed decrease in oxygen extraction fraction may be due to nonoxidative glucose metabolism during ketamine induced increase in glutamate release. PMID- 15114202 TI - Comparison of a lower-lipid propofol emulsion with the standard emulsion for sedation during monitored anesthesia care. AB - BACKGROUND: The currently used emulsion formulations of 1% propofol contain 10% soybean oil. However, a new emulsion of 1% propofol (Ampofol) containing 50% less lipid has recently become available for clinical investigation. This study was designed to compare the pharmacodynamic properties of Ampofol with those of a standard formulation (Diprivan) when administered for intraoperative sedation. METHODS: Sixty healthy outpatients undergoing minor operations with local anesthesia were randomly assigned to receive either Ampofol (n = 31) or Diprivan (n = 29) for intravenous sedation. The sedation was initiated with an intravenous loading dose of propofol, 0.75 mg/kg, followed by an initial infusion rate of 50 microg x kg(-1) x min(-1) to achieve an Observer's Assessment of Alertness/Sedation score of 3. The targeted level of sedation was maintained with a variable-rate propofol infusion during the operation. The onset times to achieving a sedation score of 3, the severity of pain on injection of the loading dose, intraoperative hemodynamic variables, and electroencephalographic Bispectral Index values were recorded. In addition, recovery times, postoperative pain and nausea, and patient satisfaction with the sedative medication were assessed. RESULTS: There were no significant differences between Ampofol and Diprivan with respect to onset times, dosage requirements, Bispectral Index values, hemodynamic variables, recovery times, or patient satisfaction scores. The incidence of moderate pain on injection was higher in the Ampofol group (26%vs. 7% with Diprivan; P < 0.05). CONCLUSIONS: Ampofol was equipotent to Diprivan with respect to its sedative properties during monitored anesthesia care. Although both groups received pretreatment with intravenous lidocaine, Ampofol was associated with more pain on injection. PMID- 15114203 TI - Molecular genetic testing for malignant hyperthermia susceptibility. AB - BACKGROUND: For more than 30 yr, the in vitro contracture test (IVCT) was the only appropriate diagnostic tool for malignant hyperthermia (MH). After the introduction of molecular genetics into MH research, guidelines for molecular genetic diagnosis of MH susceptibility were published. The aim of this study was to establish applicability of the guidelines, sensitivity, and specificity of genetic testing in MH and advantages for studied patients. METHODS: The IVCT was performed following the guidelines of the European MH Group. Mutation analyses were performed by amplification of genomic DNA by polymerase chain reaction and restriction enzyme digestion. RESULTS: Two hundred eight individuals underwent MH testing between January 2001 and April 2003. In 32 of 67 initially genetic-tested patients, the familial mutation was identified, and they were diagnosed as MH susceptible. The IVCT followed negative genetic test results in 20 patients, and all but one had negative IVCT results. Three patients were scheduled to undergo elective surgery, and IVCT and genetic testing were performed simultaneously. All three had positive IVCT results and were carriers of their familial mutation. CONCLUSIONS: In families with known MH mutations, there is a 50% chance of reliably confirming MH susceptibility by noninvasive testing. The authors found the negative predictive value of genetic testing to be 0.95 (95% confidence interval, 0.75-0.99), but for patient safety, they still recommend following the guidelines for genetic testing in MH and therefore performing an IVCT in case of negative genetic results. PMID- 15114204 TI - Moderate increase in intraabdominal pressure attenuates gastric mucosal oxygen saturation in patients undergoing laparoscopy. AB - BACKGROUND: Perioperative disturbances of microvascular blood flow and oxygenation in the intestinal tract have been hypothesized to play an important role in development of the multiple organ dysfunction syndrome. Herein, increased intra-abdominal pressure (IAP) has been identified as a key factor in the initiation of the pathophysiologic cascade. The authors hypothesized that increasing the IAP by intraperitoneal insufflation of carbon dioxide attenuates microvascular oxygen saturation in gastric mucosa. They tested this hypothesis in a prospective, observational study in 16 patients scheduled to undergo elective diagnostic laparoscopy. METHODS: The authors continuously assessed microvascular oxygen saturation in gastric mucosa by reflectance spectrophotometry. Simultaneously systemic oxygen saturation, heart rate, arterial blood pressure, and ventilation-derived variables were measured noninvasively. During general anesthesia and controlled mechanical ventilation, baseline values were obtained. Thereafter, the IAP was increased to 8 and 12 mmHg, respectively, followed by a control period after desufflation. RESULTS: The increase in IAP from baseline to 8 mmHg decreased microvascular oxygen saturation in gastric mucosa from 69+/-7% (mean +/- SD) to 63+/-8% at 8 mmHg IAP (P <0.05), with a further significant reduction to 54+/-13% at 12 mmHg IAP (P <0.01). Microvascular oxygen saturation in gastric mucosa recovered rapidly to baseline level (66 +/- 10%) after release of increased IAP. In striking contrast to regional mucosal oxygen saturation, systemic oxygenation did not change with either of the interventions. CONCLUSIONS: The results suggest that increasing intraabdominal pressure to moderate levels, commonly applied to induce a surgical pneumoperitoneum, decreases gastric mucosal oxygen saturation. PMID- 15114205 TI - Altered cell-mediated immunity and increased postoperative infection rate in long term alcoholic patients. AB - BACKGROUND: Preoperative alteration of T cell-mediated immunity as well as an altered immune response to surgical stress were found in long-term alcoholic patients. The aim of this study was to evaluate perioperative T cell-mediated immune parameters as well as cytokine release from whole blood cells after lipopolysaccharide stimulation and its association with postoperative infections. METHODS: Fifty-four patients undergoing elective surgery of the aerodigestive tract were included in this prospective observational study. Long-term alcoholic patients (n = 31) were defined as having a daily ethanol consumption of at least 60 g and fulfilling the Diagnostic and Statistical Manual of Mental Disorders for either alcohol abuse or alcohol dependence. The nonalcoholic patients (n = 23) were defined as drinking less than 60 g ethanol/day. Blood samples to analyze the immune status were obtained on morning before surgery and on the morning of days 1, 3, and 5 after surgery. RESULTS: Basic patient characteristics did not differ between groups. Before surgery, the T helper 1:T helper 2 ratio (Th1: Th2) was significantly lower (P < 0.01), whereas plasma interleukin 1beta and lipopolysaccharide-stimulated interleukin 1ra from whole blood cells were increased in long-term alcoholic patients. After surgery, a significant suppression of the cytotoxic lymphocyte ratio (Tc1:Tc2), the interferon gamma:interleukin 10 ratio from lipopolysaccharide-stimulated whole blood cells, and a significant increase of plasma interleukin 10 was observed. Long-term alcoholics had more frequent postoperative infections compared with nonalcoholic patients (54%vs. 26%; P = 0.03). CONCLUSIONS: T helper cell-mediated immunity was significantly suppressed before surgery and possibly led to inadequate cytotoxic lymphocyte and whole blood cell response in long-term alcoholic patients after surgery. This altered cell-mediated immunity might have accounted for the increased infection rate in long-term alcoholic patients after surgery. PMID- 15114206 TI - Cerebral oxygen extraction and autoregulation during extracorporeal whole body hyperthermia in humans. AB - BACKGROUND: The effects of hyperthermia on the human brain are incompletely understood. This study assessed the effects of whole body hyperthermia on cerebral oxygen extraction and autoregulation in humans. METHODS: Nineteen patients with chronic hepatitis C virus infection, not responding to interferon treatment, were subjected to experimental therapy with extracorporeal whole body hyperthermia at 41.8 degrees C for 120 min under propofol anesthesia (23 sessions total). During treatment series A (13 sessions), end-tidal carbon dioxide was allowed to increase during heating. During series B (10 sessions), end-tidal carbon dioxide was maintained approximately constant. Cerebral oxygen extraction (arterial to jugular venous difference of oxygen content) and middle cerebral artery blood flow velocity were continuously measured. Cerebral pressure-flow autoregulation was assessed by static tests using phenylephrine infusion and by assessing the transient hyperemic response to carotid compression and release. RESULTS: For treatment series A, cerebral oxygen extraction decreased 2.2-fold and cerebral blood flow velocity increased 2.0-fold during heating. For series B, oxygen extraction decreased 1.6-fold and flow velocity increased 1.5-fold. Jugular venous oxygen saturation and lactate measurements did not indicate cerebral ischemia at any temperature. Static autoregulation test results indicated loss of cerebrovascular reactivity during hyperthermia for both series A and series B. The transient hyperemic response ratio did not decrease until the temperature reached approximately 40 degrees C. Per degree Celsius temperature increase, the transient hyperemic response ratio decreased 0.07 (95% confidence interval, 0.05-0.09; P = 0.000). This association remained after adjustment for variations in arterial partial pressure of carbon dioxide, mean arterial pressure, and propofol blood concentration. CONCLUSION: Profound hyperthermia during propofol anesthesia is associated with decreased cerebral oxygen extraction, increased cerebral blood flow velocity, and impaired pressure-flow autoregulation, indicating transient partial vasoparalysis. PMID- 15114207 TI - Effect of hypertonic saline infusion on postoperative cellular immune function: a randomized controlled clinical trial. AB - BACKGROUND: Previous studies found hypertonicity to affect immune responses in intact laboratory animals and in human blood cell cultures. In this study, the authors investigated the cellular immune response to surgery after preoperative infusion of hypertonic saline in humans. METHODS: Sixty-two women scheduled to undergo abdominal hysterectomy were randomly assigned to single-blinded infusion of 4 ml/kg NaCl, 7.5%; 4 ml/kg NaCl, 0.9%; or 32 ml/kg NaCl, 0.9%, over 20 min. Blood was collected at baseline, during surgery, and 1, 24, and 48 h after surgery for the determination of leukocyte and differential counts, flow cytometric phenotyping of mononuclear cells, and natural killer cell activity against K 562 tumor cells. Phytohemagglutinin-induced lymphocyte proliferation, plasma elastase, and neutrophil chemotaxis were measured at the same time points except during surgery. The authors tested cell-mediated immune function in vivo by delayed-type hypersensitivity reaction in the skin. RESULTS: Surgery induced well-known changes in the cellular immune response, which were unrelated to the tonicity or volume of the infused fluids. CONCLUSION: Infusion of a clinically relevant dose of hypertonic saline did not seem to modify the postoperative cellular immune response after elective abdominal hysterectomy. PMID- 15114208 TI - Can acceleromyography detect low levels of residual paralysis? A probability approach to detect a mechanomyographic train-of-four ratio of 0.9. AB - BACKGROUND: The incidence of residual paralysis, i.e., a mechanomyographic train of-four (TOF) ratio (T4/T1) less than 0.9, remains frequent. Routine acceleromyography has been proposed to detect residual paralysis in clinical practice. Although acceleromyographic data are easy to obtain, they differ from mechanomyographic data, with which they are not interchangeable. The current study aimed to determine (1) the acceleromyographic TOF ratio that detects residual paralysis with a 95% probability, and (2) the impact of calibration and normalization on this predictive acceleromyographic value. METHODS: In 60 patients, recovery from neuromuscular block was assessed simultaneously with mechanomyography and acceleromyography. To obtain calibrated acceleromyographic TOF ratios in group A, the implemented calibration modus 2 was activated in the TOF-Watch S; to obtain uncalibrated acceleromyographic TOF ratios in group B, the current was manually set at 50 mA (n = 30 for each). In addition, data in group B were normalized (i.e., dividing the final TOF ratio by the baseline value). The agreement between mechanomyography and acceleromyography was assessed by calculating the intraclass correlation coefficient. Negative predictive values were calculated for detecting residual paralysis from acceleromyographic TOFs of 0.9, 0.95, and 1.0. RESULTS: GROUP A: : For a mechanomyographic TOF of 0.9 or greater, the corresponding acceleromyographic TOF was 0.95 (range, 0.86-1.0), and the negative predictive values for acceleromyographic TOFs of 0.9, 0.95, and 1.0 were 37% (95% CI, 20-56%), 70% (95% CI, 51-85%), and 97% (95% CI, 83-100%), respectively. Group B: Without normalization, an acceleromyographic TOF of 0.97 (range, 0.68-1.18) corresponded to a mechanomyographic TOF of 0.9 or greater, with negative predictive values for acceleromyographic TOFs of 0.9, 0.95, and 1.0 being 40% (95% CI, 23-59%), 60% (95% CI, 41-77%), and 77% (95% CI, 58-90%), respectively. After normalization, an acceleromyographic TOF of 0.89 (range, 0.63 1.06) corresponded to a mechanomyographic TOF of 0.9 or greater, and the negative predictive values of acceleromyographic TOFs of 0.9, 0.95, and 1.0 were 89% (95% CI, 70-98%), 92% (95% CI, 75-99%), and 96% (95% CI, 80-100%), respectively. CONCLUSION: To exclude residual paralysis reliably when using acceleromyography, TOF recovery to 1.0 is mandatory. PMID- 15114209 TI - Smoking behavior and perceived stress in cigarette smokers undergoing elective surgery. AB - BACKGROUND: The forced abstinence from cigarettes accompanying surgery in smoke free facilities may increase psychological stress by removing a coping mechanism and by nicotine withdrawal. The authors tested the hypothesis that abstinence from cigarette smoking contributes to psychological stress in the perioperative period. METHODS: The authors assessed measures of nicotine withdrawal (Hughes Hatsukami nicotine withdrawal scale) and perceived stress (including the Perceived Stress Scale) in 141 cigarette smokers scheduled to undergo elective surgery. To separate the effects of stress arising from tobacco abstinence from the effects of other perioperative stressors, such as pain, these measures were also obtained in 150 surgical patients who did not use tobacco. Assessments were performed at intervals beginning at the time of preoperative medical evaluation and ending 30 days postoperatively. RESULTS: Perceived Stress Scale scores were significantly (P < 0.001) higher in smokers throughout the study period. There was little significant interaction between smoking status and time, indicating that changes in Perceived Stress Scale score during the perioperative period did not differ between smokers and nonsmokers. The same result was found if analysis was restricted to data collected before hospital discharge (and thus during assured abstinence). Similar results were found for the nicotine withdrawal scale, suggesting that smokers did not experience more withdrawal symptoms relative to nonsmokers. CONCLUSIONS: Although smokers report increased baseline stress, smoking status does not affect changes in perceived stress over the perioperative period. Nicotine withdrawal symptoms do not seem to be a clinically significant problem in the perioperative period for most smokers. PMID- 15114210 TI - Development and psychometric evaluation of the pediatric anesthesia emergence delirium scale. AB - BACKGROUND: Emergence delirium has been investigated in several clinical trials. However, no reliable and valid rating scale exists to measure this phenomenon in children. Therefore, the authors developed and evaluated the Pediatric Anesthesia Emergence Delirium (PAED) scale to measure emergence delirium in children. METHODS: A list of scale items that were statements describing the emergence behavior of children was compiled, and the items were evaluated for content validity and statistical significance. Items that satisfied these evaluations comprised the PAED scale. Each item was scored from 1 to 4 (with reverse scoring where applicable), and the scores were summed to obtain a total scale score. The degree of emergence delirium varied directly with the total score. Fifty children were enrolled to determine the reliability and validity of the PAED scale. Scale validity was evaluated using five hypotheses: The PAED scale scores correlated negatively with age and time to awakening and positively with clinical judgment scores and Post Hospital Behavior Questionnaire scores, and were greater after sevoflurane than after halothane. The sensitivity of the scale was also determined. RESULTS: Five of 27 items that satisfied the content validity and statistical analysis became the PAED scale: (1) The child makes eye contact with the caregiver, (2) the child's actions are purposeful, (3) the child is aware of his/her surroundings, (4) the child is restless, and (5) the child is inconsolable. The internal consistency of the PAED scale was 0.89, and the reliability was 0.84 (95% confidence interval, 0.76-0.90). Three hypotheses supported the validity of the scale: The scores correlated negatively with age (r = -0.31, P <0.04) and time to awakening (r = -0.5, P <0.001) and were greater after sevoflurane anesthesia than halothane (P <0.008). The sensitivity was 0.64. CONCLUSIONS: These results support the reliability and validity of the PAED scale. PMID- 15114211 TI - Unanticipated difficult airway in anesthetized patients: prospective validation of a management algorithm. AB - BACKGROUND: Management strategies conceived to improve patient safety in anesthesia have rarely been assessed prospectively. The authors undertook a prospective evaluation of a predefined algorithm for unanticipated difficult airway management. METHODS: After a 2-month period of training in airway management, 41 anesthesiologists were asked to follow a predefined algorithm for management in the case of an unanticipated difficult airway. Two different scenarios were distinguished: "cannot intubate" and "cannot ventilate." The gum elastic bougie and the Intubating Laryngeal Mask Airway (ILMA) were proposed as the first and second steps in the case of impossible laryngoscope-assisted tracheal intubation, respectively. In the case of impossible ventilation or difficult ventilation, the IMLA was recommended, followed by percutaneous transtracheal jet ventilation. The patient's details, adherence rate to the algorithm, efficacy, and complications of airway management processes were recorded. RESULTS: Impossible ventilation never occurred during the 18-month study. One hundred cases of unexpected difficult airway were recorded (0.9%) among 11,257 intubations. Deviation from the algorithm was recorded in three cases, and two patients were wakened before any alternative intubation technique attempt. All remaining patients were successfully ventilated with either the facemask (89 of 95) or the ILMA (6 of 95). Six difficult-ventilation patients required the ILMA before completion of the first intubation step. Eighty patients were intubated with the gum elastic bougie, and 13 required a blind intubation through the ILMA. Two patients ventilated with the ILMA were never intubated. CONCLUSION: When applied in accordance with a predefined algorithm, the gum elastic bougie and the ILMA are effective to solve most problems occurring during unexpected difficult airway management. PMID- 15114212 TI - Propofol neuroprotection in cerebral ischemia and its effects on low-molecular weight antioxidants and skilled motor tasks. AB - BACKGROUND: Propofol is neuroprotective when administered immediately after stroke. The therapeutic window, duration of administration, and antioxidant mechanisms of propofol in neuroprotection are not known. The effects of propofol after stroke were examined in the conscious animal. The authors have previously shown that light propofol anesthesia (25 mg x kg(-1) x h(-1)) for a period of 4 h, even if delayed 1 h after the onset of ischemia, decreases infarct volume 3 days after the stroke. METHODS: Cerebral ischemia was induced in awake Wistar rats by a local intracerebral injection of the potent vasoconstrictor, endothelin (6 pmol in 3 microl) into the striatum. Propofol treatment after ischemia was delayed up to 4 h, and the infusion period shortened from 4 h to 1 h. Infarct volume was assessed 3 or 21 days after the stroke. Neurologic outcome was evaluated on days 14-21 after ischemia. Tissue ascorbate and glutathione concentrations were evaluated at 4 h and 3 days after ischemia. RESULTS: Infarct volumes were reduced 3 days after ischemia when propofol treatment (25 mg x kg( 1) x h(-1)) was delayed for 2 h (0.5+/-0.3 mm3) but not 4 h (2.0+/-0.9 mm3), compared with intralipid controls (2.4 +/- 0.7 mm3). The propofol infusion period of 3 h but not 1 h reduced infarct volume. Propofol treatment did not reduce infarct volume 21 days after the stroke, although motor function improvements (Montoya staircase test) were observed 14-21 days after the stroke. Propofol neuroprotection was independent of tissue ascorbate and glutathione concentrations. CONCLUSIONS: Concurrent or delayed administration of propofol is neuroprotective 3 days after ischemia. Although there were no differences in infarct volume 21 days after ischemia, propofol-treated animals had functional improvements at this time. PMID- 15114213 TI - Effects of isoflurane versus fentanyl-nitrous oxide anesthesia on long-term outcome from severe forebrain ischemia in the rat. AB - BACKGROUND: This study examined long-term outcome from severe forebrain ischemia in the rat, as a function of anesthetic given during the ischemic injury. METHODS: Rats were subjected to 10 min of near-complete forebrain ischemia while anesthetized with either 1.4% isoflurane or 70% nitrous oxide-fentanyl. Neurologic and histologic outcomes were measured at 5 days, 3 weeks, or 3 months after ischemia. RESULTS: At 5 days, isoflurane-anesthetized rats had less damage than did fentanyl-nitrous oxide-anesthetized rats (mean +/- SD, percent alive hippocampal CA1 neurons = 58+/-29 vs. 20+/-16, respectively; P = 0.011). This was accompanied by improved motor function in the isoflurane group (P = 0.002). At 3 weeks, there was no difference between groups for either outcome variable (percent alive CA1 neurons = 35+/-26 and 36+/-28 for isoflurane and fentanyl nitrous oxide, respectively). Similarly, at 3 months, there was no difference between groups (percent alive CA1 neurons = 56+/-27 and 60+/-27 for isoflurane and fentanyl-nitrous oxide, respectively). Morris water maze performance at 3 months was similar between anesthetic groups and was also similar to sham performance. The percent alive CA1 neurons in the fentanyl-nitrous oxide group increased with duration of recovery (P = 0.004). There were no differences among isoflurane groups over time (5 days vs. 3 weeks, P = 0.26; 5 days vs. 3 months, P = 0.99; 3 week vs. 3 months, P = 0.32). CONCLUSIONS: This study found no change in the percent alive CA1 hippocampal neurons as a function of duration of recovery from severe forebrain ischemia in isoflurane anesthetized rats. In contrast, the percent alive CA1 neurons in fentanyl-nitrous oxide-anesthetized rats tripled over 3 months of recovery. The natural history of long-term responses to forebrain ischemia requires further study before conclusions can be drawn with respect to the permanence of isoflurane neuroprotection. PMID- 15114214 TI - Laser ignition of surgical drape materials in air, 50% oxygen, and 95% oxygen. AB - BACKGROUND: Operating room fires fueled by surgical drapes and ignited by high energy surgical tools in air and oxygen-enriched atmospheres continue to occur. METHODS: The authors examined the time to ignition of huck towels and three commonly used surgical drape materials in air, 50% oxygen, and 95% oxygen using a carbon dioxide surgical laser as an ignition source. In addition, a phenol polymer fabric was tested. RESULTS: In air, polypropylene and phenol polymer do not ignite. For polypropylene, the laser instantly vaporized a hole, and therefore, interaction between the laser and material ceased. When tested in combination with another material, the polypropylene time to ignition assumed the behavior of the material with which it was combined. For phenol polymer, the laser did not penetrate the material. Huck towels, cotton-polyester, and non woven cellulose-polyester ignited in air with decreasing times to ignition. All tested materials ignited in 50% and 95% oxygen. CONCLUSION: The results of this study reveal that with increasing oxygen concentration, the time to ignition becomes shorter, and the consequences become more severe. The possibility exists for manufacturers to develop drape materials that are safer than existing materials. PMID- 15114215 TI - Effects of anesthesia and recovery from ketamine racemate and enantiomers on regional cerebral glucose metabolism in rats. AB - BACKGROUND: Unlike most anesthetics, ketamine racemate (S, R (+/-)-ketamine) induces heterogenous changes in cerebral metabolism. S, R (+/-)-ketamine is an equimolar mixture of two enantiomers, S (+)-ketamine and R (-)-ketamine, which differ in affinity for neuroreceptors and pharmacologic activities. This study investigated comparatively the effects of ketamine racemate and enantiomers on cerebral metabolism. METHODS: Regional cerebral metabolic rates for glucose (rCMRglc) were determined with the quantitative, autoradiographic [C]2-deoxy-d glucose technique in 40 brain regions of Fischer-344 rats. rCMRglc were measured in three groups of rats during equimolar anesthesia, 10 min after intraperitoneal injection of 170 mg/kg S, R (+/-)-ketamine, S (+)-ketamine, or R (-)-ketamine; in three groups of rats during recovery from equivalent anesthesia, 20 min after intravenous injection of 20, 12.5, and 30 mg/kg S, R (+/-)-ketamine, S (+) ketamine, or R (-)-ketamine; and in two groups of saline-injected control rats. RESULTS: S, R (+/-)-ketamine and S (+)-ketamine induced a sustained anesthesia; deep rCMRglc decreases in 22 and 14 cortical, thalamic, cerebellar, and brainstem regions; and rCMRglc increases in two limbic regions (average decreases, 23 and 15%). R (-)-ketamine determined a shorter anesthesia, lesser rCMRglc decreases in 11 brain areas, and marked rCMRglc increases in 14 basal ganglia and limbic regions (average decrease, 4%). S, R (+/-)-ketamine, S (+)-ketamine, and R (-) ketamine all produced postanesthetic behavioral activation; widespread rCMRglc increases in 28, 16, and 20 cortical, thalamic, basal ganglia, limbic, and brainstem regions; and rCMRglc decreases in few auditory and limbic regions (average increases, 35, 13, and 20%). CONCLUSIONS: S, R (+/-)-ketamine and S (+) ketamine anesthesia but not R (-)-ketamine anesthesia induced widespread rCMRglc reductions that were unreported but are typical of gaseous and intravenous general anesthetics. Postanesthetic recovery led to divergent, sharp behavioral and rCMRglc activations. The relation to dose of behavioral and rCMRglc effects differs from those of aminergic agents and resembles those of N-methyl-d aspartate receptor antagonists, suggesting that ketamine racemate and enantiomers may preferentially interact with this receptor type. PMID- 15114216 TI - Myocardial effects of halothane and sevoflurane in diabetic rats. AB - BACKGROUND: Diabetes induces significant myocardial abnormalities, but the effects of halogenated anesthetics on this diseased myocardium remain a matter of debate. METHODS: Left ventricular papillary muscles and triton-skinned cardiac fibers were provided from control and streptozotocin-induced diabetic rats. The effects of halothane and sevoflurane were studied on inotropic and lusitropic responses, under low (isotony) and high (isometry) loads in papillary muscles and then on isometric tension-Ca2+ concentration (pCa) relations obtained in triton skinned cardiac fibers. Data are presented as mean +/- SD. RESULTS: Sevoflurane and halothane induced a negative inotropic effect that was more important in diabetic rats (active force: 1.5% halothane, 19+/-6 vs. 24+/-6% of baseline, P < 0.05; 3.6% sevoflurane, 47+/-14 vs. 69+/-17% of baseline, P < 0.05). However, when differences in minimum alveolar concentration were considered, no significant difference was observed between groups for halothane. The effects of halothane and sevoflurane on isotonic relaxation and postrest potentiation were not significantly different between groups. In contrast, the decrease in Ca myofilament sensitivity produced by each anesthetic agent was greater in diabetic rats than in control rats (0.65% halothane, -0.15+/-0.07 vs. -0.05+/-0.04 pCa unit, P < 0.05; 1.8% sevoflurane, -0.12+/-0.06 vs. -0.06+/-0.04 pCa unit, P < 0.05). CONCLUSIONS: The negative inotropic effect of halothane and sevoflurane was greater in diabetic rats, mainly because of a significant decrease in myofilament Ca sensitivity. PMID- 15114217 TI - Effects of dopamine, dobutamine, and dopexamine on microcirculatory blood flow in the gastrointestinal tract during sepsis and anesthesia. AB - BACKGROUND: Insufficient blood flow to the splanchnic organs is believed to be an important contributory factor for the development of organ failure after septic shock. It has been suggested that increasing systemic flow also may improve splanchnic blood flow in septic patients. The aim of this study was to compare the effects of three commonly used inotropic agents, dopamine, dobutamine, and dopexamine, on systemic (cardiac index), regional (superior mesenteric artery), and local (micro-circulatory) blood flow during septic shock in pigs. METHODS: Eight pigs were intravenously anesthetized, mechanically ventilated, and exposed to sepsis induced by fecal peritonitis. Cardiac index was measured with thermodilution, superior mesenteric artery flow was measured with ultrasound transit time flowmetry, and microcirculatory blood flow was continuously measured with a six-channel laser Doppler flowmetry in the gastric, jejunal, and colon mucosa as well as in the kidney, pancreas, and jejunal muscularis. Each animal received, in a random-order, crossover design, the three test drugs, one at a time: 5 and 10 microg x kg(-1) x min(-1) dopamine, 5 and 10 microg x kg(-1) x min(-1) dobutamine, and 1 and 2 microg x kg(-1) x min(-1) dopexamine. Administration of each drug at each dose continued for 30 min and was followed by a 40- to 60-min recovery period. A new baseline was taken before the next drug was administered. RESULTS: All three drugs significantly increased cardiac index; dopamine by 18%, dobutamine by 48%, and dopexamine by 35%, compared with baseline (P < 0.001 for each). At the same time, superior mesenteric artery flow increased by 33% (P < 0.01) with dopamine and 13% (P < 0.01) with dopexamine, whereas it did not change with dobutamine. Microcirculatory blood flow did not change significantly in any of the organs studied with any of the drugs tested. CONCLUSION: All the inotropic agents markedly increased cardiac output in this sepsis model. However, increased systemic flow did not reach the microcirculation in the gastrointestinal tract. This may in part explain why some of the clinical trials, in which systemic oxygen delivery was deliberately increased by administration of inotropic drugs, have failed to improve survival in critically ill patients. PMID- 15114218 TI - Propofol modulates gamma-aminobutyric acid-mediated inhibitory neurotransmission to cardiac vagal neurons in the nucleus ambiguus. AB - BACKGROUND: Although it is well recognized that anesthetics modulate the central control of cardiorespiratory homeostasis, the cellular mechanisms by which anesthetics alter cardiac parasympathetic activity are poorly understood. One common site of action of anesthetics is inhibitory neurotransmission. This study investigates the effect of propofol on gamma-aminobutyric acid-mediated (GABAergic) and glycinergic neurotransmission to cardiac parasympathetic neurons. METHODS: Cardiac parasympathetic neurons were identified in vitro by the presence of a retrograde fluorescent tracer, and spontaneous GABAergic and glycinergic synaptic currents were examined using whole cell patch clamp techniques. RESULTS: Propofol at concentrations of 1.0 microm and greater significantly (P < 0.05) increased the duration and decay time of spontaneous GABAergic inhibitory postsynaptic currents. To determine whether the action of propofol was at presynaptic or postsynaptic sites, tetrodotoxin was applied to isolate miniature inhibitory postsynaptic currents. Propofol at concentrations of 1.0 microm and greater significantly (P < 0.05) prolonged the decay time and duration of miniature inhibitory postsynaptic currents, indicating that propofol directly alters GABAergic neurotransmission at a postsynaptic site. Propofol at high concentrations (> or =50 microm) also inhibited the frequency of both GABAergic inhibitory postsynaptic currents and miniature inhibitory postsynaptic currents. Propofol at concentrations up to 50 microm had no effect on glycinergic neurotransmission. CONCLUSIONS: Propofol may vary heart rate by modulating GABAergic neurotransmission to cardiac parasympathetic neurons. At clinically relevant concentrations (> or =1.0 microm), propofol facilitated GABAergic responses in cardiac vagal neurons by increasing decay time, which would increase inhibition of cardioinhibitory cardiac vagal neurons and evoke an increase in heart rate. At higher supraclinical concentrations (> or =50 microm), propofol inhibits GABAergic neurotransmission to cardiac vagal neurons, which would evoke a decrease in heart rate. PMID- 15114219 TI - Lidocaine inhibits tyrosine kinase activity of the epidermal growth factor receptor and suppresses proliferation of corneal epithelial cells. AB - BACKGROUND: Although lidocaine is recognized as an excellent topical corneal analgesic, its toxic effect on corneal epithelial cells limits its use during corneal epithelial wound healing. Mechanism of the impairment of corneal reepithelialization with lidocaine, however, has not been evaluated. The authors' previous study revealed that lidocaine inhibits the activity of tyrosine kinase receptors through the interaction with specific amino acid sequences around autophosphorylation sites, including acidic, basic, and aromatic amino acids. Epidermal growth factor receptor (EGFR), a tyrosine kinase receptor with an important role in epithelial cell proliferation after corneal wounding, also possesses these amino acids sequences around autophosphorylation sites. The authors hypothesized that lidocaine would suppress tyrosine kinase activity of EGFR and would impair corneal epithelial cell proliferation. METHODS: To investigate the effect of lidocaine (4 microM-40 mM) on epidermal growth factor (EGF)-stimulated autophosphorylation of EGFR, the authors studied purified EGFR in microtubes. They cultured human corneal epithelial cells (HCECs) with EGF and lidocaine to investigate the effect of lidocaine on cell proliferation and on autophosphorylation of EGFR in HCECs. RESULTS: Lidocaine (> or =400 microM) significantly suppressed EGF-stimulated autophosphorylation of the purified EGFR. In the HCEC study, EGF alone stimulated cell proliferation and increased autophosphorylation of EGFR in HCECs. Lidocaine (> or = 400 microM) significantly suppressed both the proliferation of HCECs promoted by EGF and EGF-stimulated autophosphorylation of EGFR. CONCLUSION: Lidocaine directly inhibits tyrosine kinase activity of EGFR and suppresses the corneal epithelial cell proliferation. PMID- 15114220 TI - Endothelial glycocalyx as an additional barrier determining extravasation of 6% hydroxyethyl starch or 5% albumin solutions in the coronary vascular bed. AB - BACKGROUND: The impact on the endothelial glycocalyx for the extravasation of colloidal infusion solutions has not been investigated sufficiently. METHODS: Isolated guinea pig hearts were perfused with Krebs-Henseleit buffer in a Langendorff mode. Solutions of 0.9% saline, 5% albumin (70 kd), or 6% hydroxyethyl starch (200 kd) were infused into the coronary system for 20 min at a rate of one third of the coronary flow, also during reperfusion after 15 min of ischemia, and after enzymatic digestion of the endothelial glycocalyx by heparinase. Net coronary fluid filtration was assessed directly by measuring the formation of transudate on the epicardial surface, and solute extravasation was assessed by measuring albumin and hydroxyethyl starch in the coronary effluent and transudate. Hearts were perfusion fixed to visualize the endothelial glycocalyx using transmission electron microscopy. RESULTS: Only infusion of hydroxyethyl starch, not infusion of albumin, significantly decreased net coronary fluid filtration. Heparinase application without ischemia increased coronary leak by 25% but did not accelerate the passage of colloids. Ischemia alone did not alter permeability. However, there was a large (approximately +200%), transient (approximately 4 min) increase in permeability for water, albumin, and hydroxyethyl starch after ischemia with heparinase application. Also, histamine (10 m) only increased permeability after pretreatment of the hearts with heparinase. The thickness of the glycocalyx after colloid administration was 0.2-0.3 microm. No glycocalyx could be detected after application of heparinase. CONCLUSION: The endothelial glycocalyx acts as a competent barrier for water and colloids. Only after its destruction do changes in endothelial morphology (postischemic reperfusion or histamine application) become effective determinants of coronary extravasation. PMID- 15114221 TI - Isoflurane differentially modulates medullary on and off neurons while suppressing hind-limb motor withdrawals. AB - BACKGROUND: Isoflurane acts primarily in the spinal cord to block movement; however, it is unclear how supraspinal sites might contribute to anesthetic effects on quantified parameters of movement such as force. METHODS: The authors investigated the effects of isoflurane on spontaneous and noxious heat-evoked activity of nociceptive reflex-modulating ON and OFF cells in the rostral ventromedial medulla of rats. Single ON or OFF neurons were recorded simultaneously with hind-limb withdrawal force elicited by graded noxious thermal hind paw stimulation. Isoflurane concentrations were administered in reference to each animal's minimum alveolar concentration (MAC) of isoflurane. RESULTS: From 0.65 to 1.15 MAC, isoflurane dose-dependently reduced spontaneous activity of ON cells by 70% (P < 0.001). OFF-cell spontaneous activity was dose-dependently increased 138% (P < 0.001). ON-cell heat-evoked activity was depressed 95% by isoflurane from 0.65 to 1.15 MAC (P < 0.001). Isoflurane-induced changes in ON- and OFF-cell activity paralleled similar reductions in withdrawal force, with the largest change in both neuronal activity and withdrawal force occurring between 0.85 and 1.15 MAC. For the lowest stimulus temperature, excitatory responses of ON cells and inhibitory responses of OFF cells were significantly greater for trials in which withdrawals occurred than for trials in which no withdrawal occurred, suggesting that responses in both classes of neurons were related to movement rather than the stimulus alone. CONCLUSIONS: The results show that isoflurane modulation of ON- and OFF-cell activity corresponds to anesthetic induced reductions in hind-limb withdrawal force, and therefore, the effects of isoflurane on these classes of neurons in rostral ventromedial medulla might contribute to motor depression. PMID- 15114222 TI - Role of lipid in sulfite-dependent propofol dimerization. AB - BACKGROUND: During long-term intravenous infusions, sulfite in sulfite-containing propofol emulsions can cause the peroxidation of lipid and dimerization of propofol. This study evaluated the role of lipid in sulfite-dependent propofol dimerization by determining the effects of individual fatty acids in soybean oil emulsion and peroxidized lipids in a model system. METHODS: Individual fatty acids, stearic (18:0), oleic (18:1), linoleic (18:2), linolenic (18:3), and arachidonic (20:4), were added to sulfite-containing propofol emulsion and incubated for 90 min at 37 degrees C. Model systems containing soybean oil (100 microl), water (900 microl), propofol (10 mg/ml), and sulfite (0.25 mg/ml) composed of oils with different peroxide values were allowed to react for 60 min at room temperature. After the reactions, propofol dimer and propofol dimer quinone were analyzed by reversed-phase high-pressure liquid chromatography. RESULTS: Propofol did not dimerize when added to aqueous sulfite unless soybean oil was also included. The addition of the polyunsaturated fatty acids (linoleic, linolenic, arachidonic) to sulfite-containing propofol emulsion resulted in large increases of propofol dimerization compared with stearic or oleic acid. Using biphasic mixtures of soybean oil and aqueous sulfite, propofol dimerization increased with increasing peroxide content of the oil. In propofol emulsion, lipoxidase and ferrous iron in the absence of sulfite also caused the dimerization of propofol. CONCLUSIONS: These results show that lipid can play a significant role in sulfite-dependent propofol dimerization. The relation of dimerization to polyunsaturated fatty acid and soybean oil peroxide content suggests that sulfite reacts with unsaturated lipid or peroxide-modified lipid to facilitate propofol dimerization. PMID- 15114223 TI - Ultrasound-guided lumbar facet nerve block: a sonoanatomic study of a new methodologic approach. AB - BACKGROUND: Lumbar facet nerve (medial branch) block for pain relief in facet syndrome is currently performed under fluoroscopic or computed tomography scan guidance. In this three-part study, the authors developed a new ultrasound-guided methodology, described the necessary landmarks and views, assessed ultrasound derived distances, and tested the clinical feasibility. METHODS: (1) A paravertebral cross-axis view and long-axis view were defined under high resolution ultrasound (15 MHz). Three needles were guided to the target point at L3-L5 in a fresh, nonembalmed cadaver under ultrasound (2-6 MHz) and were subsequently traced by means of dissection. (2) The lumbar regions of 20 volunteers (9 women, 11 men; median age, 36 yr [23-67 yr]; median body mass index, 23 kg/m2 [19-36 kg/m2]) were studied with ultrasound (3.5 MHz) to assess visibility of landmarks and relevant distances at L3-L5 in a total of 240 views. (3) Twenty-eight ultrasound-guided blocks were performed in five patients (two women, three men; median age, 51 yr [31-68 yr]) and controlled under fluoroscopy. RESULTS: In the cadaver, needle positions were correct as revealed by dissection at all three levels. In the volunteers, ultrasound landmarks were delineated as good in 19 and of sufficient quality in one (body mass index, 36 kg/m2). Skin target distances increased from L3 to L5, reaching statistical significance (*, **P < 0.05) between these levels on both sides: L3r, 45+/-6 mm*; L4r, 48+/-7 mm; L5r, 50+/-6 mm*; L3l, 44+/-5 mm**; L4l, 47+/-6 mm; L5l, 50+/-6 mm**. In patients, 25 of 28 ultrasound-guided needles were placed accurately, with the remaining three closer than 5 mm to the radiologically defined target point. CONCLUSION: Ultrasound guidance seems to be a promising new technique with clinical relevance and the potential to increase practicability while avoiding radiation in lumbar facet nerve block. PMID- 15114224 TI - Effect of methylprednisolone on neuropathic pain and spinal glial activation in rats. AB - BACKGROUND: Basic data are lacking regarding the efficacy and mechanisms of action of corticosteroids in neuropathic pain. Because recent studies indicate that spinal glial activation mediates the pathologic pain states, the authors sought to determine the effects of systemic and intrathecal methylprednisolone on the development and maintenance of neuropathic pain and spinal glial activation in a rat model. METHODS: Rats were anesthetized, and L5 and L6 spinal nerves were tightly ligated. Then, continuous infusion of systemic (4 mg x kg(-1) x day(-1)) or intrathecal (80 microg x kg(-1) x day(-1)) methylprednisolone or saline was started. Mechanical allodynia and thermal hyperalgesia were evaluated on days 4 and 7 postoperatively with von Frey and Hargreaves tests, respectively. Spinal astrocytic activation was evaluated with glial fibrillary acidic protein immunoreactivity on day 7. In other groups of rats, continuous 3-day treatment with intrathecal methylprednisolone or saline was started 7 days after spinal nerve ligation, when neuropathic pain had already developed. Behavioral tests and immunostaining were performed up to 3 weeks after the treatment. RESULTS: Spinal nerve ligation induced mechanical allodynia and thermal hyperalgesia on days 4 and 7 postoperatively. Glial fibrillary acidic protein immunoreactivity was remarkably enhanced on day 7. Both systemic and intrathecal methylprednisolone inhibited the development of neuropathic pain states and glial activation. Three day treatment with intrathecal methylprednisolone reversed existing neuropathic pain state and glial activation up to 3 weeks after the treatment. CONCLUSION: : Systemic and intrathecal methylprednisolone inhibited spinal glial activation and the development and maintenance of a neuropathic pain state in a rat model of spinal nerve ligation. PMID- 15114225 TI - Spinal adenosine receptor activation reduces hypersensitivity after surgery by a different mechanism than after nerve injury. AB - BACKGROUND: Intrathecal adenosine has antinociceptive effects under conditions of hypersensitivity. T62 (2-amino-3-(4-chlorobenzoyl)-5,6,7,8 tetrahydrobenzothiophen) is an allosteric adenosine receptor modulator that enhances adenosine binding to the A1 receptor. Intrathecal T62 reduces hypersensitivity to mechanical stimuli in a rat model of neuropathic pain by a circuit that totally relies on activation of alpha2 adrenoceptors. Here, the authors tested whether this same dependence was present in the acute setting of hypersensitivity after surgery. METHODS: Intrathecal catheters were inserted in male Sprague-Dawley rats. An incision of the plantar aspect of the hind paw resulted 24 h later in hypersensitivity, as measured by applying von Frey filaments to the paw. At this time, rats received intrathecal T62, clonidine, or the combination in a blinded, isobolographic design. The effect of the alpha2 adrenoceptor antagonist idazoxan on T62 was also tested. RESULTS: Intrathecal T62 produced a dose-dependent antihypersensitivity effect, with no effect on ambulation or activity level. Clonidine also produced a dose-dependent antihypersensitivity effect. The ED40 (95% confidence interval) for T62 was 0.77 (0.63-0.91) microg, and that for clonidine was 1.23 (0.56-1.9) microg. Isobolographic analysis indicated synergism between T62 and clonidine. Intrathecal pretreatment with idazoxan only partially inhibited the antihypersensitivity effect of T62. CONCLUSIONS: Intrathecal T62 is effective for postoperative hypersensitivity. The synergy of T62 with clonidine and its only partial antagonism by idazoxan suggest that T62 does not rely entirely on activation of alpha2 adrenoceptors. These results indicate that, after surgery, T62 acts via a mechanism different from that of spinal nerve ligation, a model of chronic neuropathic pain. PMID- 15114226 TI - Experimental pain models reveal no sex differences in pentazocine analgesia in humans. AB - BACKGROUND: Accumulating evidence suggests that there are sex differences in analgesic responses to opioid agonists. Several studies using an oral surgery pain model have reported more robust analgesia to kappa-agonist-antagonists (e.g., pentazocine, nalbuphine, butorphanol) among women than among men. However, evidence of sex differences in kappa-agonist-antagonist effects from studies of experimentally induced pain in humans is lacking. METHODS: Therefore, the analgesic effects of intravenous pentazocine (0.5 mg/kg) were determined in healthy women (n = 41) and men (n = 38) using three experimental pain models: heat pain, pressure pain, and ischemic pain. Each pain procedure was conducted before and after double-blind administration of both pentazocine and saline, which occurred on separate days in counterbalanced order. RESULTS: Compared with saline, pentazocine produced significant analgesic responses for all pain stimuli. However, no sex differences in pentazocine analgesia emerged. Effect sizes for the sex differences were computed; the magnitude of effects was small, and an equal number of measures showed greater analgesia in men than in women. Also, analgesic responses were not highly correlated across pain modalities, suggesting that different mechanisms may underlie analgesia for disparate types of pain. CONCLUSIONS: These findings indicate significant analgesic responses to pentazocine in both men and women across multiple experimental pain assays, and the absence of sex differences contrasts with previous data from the oral surgery model. The most likely explanation for the discrepancy in results is that of differences in the pain assays. These findings are important because they suggest that sex differences in opioid analgesia may be specific to certain types of pain. PMID- 15114227 TI - Queuing theory accurately models the need for critical care resources. AB - BACKGROUND: Allocation of scarce resources presents an increasing challenge to hospital administrators and health policy makers. Intensive care units can present bottlenecks within busy hospitals, but their expansion is costly and difficult to gauge. Although mathematical tools have been suggested for determining the proper number of intensive care beds necessary to serve a given demand, the performance of such models has not been prospectively evaluated over significant periods. METHODS: The authors prospectively collected 2 years' admission, discharge, and turn-away data in a busy, urban intensive care unit. Using queuing theory, they then constructed a mathematical model of patient flow, compared predictions from the model to observed performance of the unit, and explored the sensitivity of the model to changes in unit size. RESULTS: The queuing model proved to be very accurate, with predicted admission turn-away rates correlating highly with those actually observed (correlation coefficient = 0.89). The model was useful in predicting both monthly responsiveness to changing demand (mean monthly difference between observed and predicted values, 0.4+/ 2.3%; range, 0-13%) and the overall 2-yr turn-away rate for the unit (21%vs. 22%). Both in practice and in simulation, turn-away rates increased exponentially when utilization exceeded 80-85%. Sensitivity analysis using the model revealed rapid and severe degradation of system performance with even the small changes in bed availability that might result from sudden staffing shortages or admission of patients with very long stays. CONCLUSIONS: The stochastic nature of patient flow may falsely lead health planners to underestimate resource needs in busy intensive care units. Although the nature of arrivals for intensive care deserves further study, when demand is random, queuing theory provides an accurate means of determining the appropriate supply of beds. PMID- 15114228 TI - Drugs and human memory (part 2). Clinical, theoretical, and methodologic issues. PMID- 15114229 TI - Electrophysiologic testing for the diagnosis of peripheral nerve injuries. PMID- 15114230 TI - Local anesthetic infusion pump systems adverse events reported to the Food and Drug Administration. PMID- 15114231 TI - Acute cardiovascular instability during percutaneous ethanol injection of a hepatocellular carcinoma under general anesthesia. PMID- 15114232 TI - Posterior inferior cerebellar artery infarction: an unusual complication of posterior spinal fusion surgery in an adolescent with idiopathic scoliosis. PMID- 15114233 TI - Dysphagia, obstructive sleep apnea, and difficult fiberoptic intubation secondary to diffuse idiopathic skeletal hyperostosis. PMID- 15114234 TI - Xenon exerts age-independent antinociception in Fischer rats. PMID- 15114235 TI - Time to maximum effect site concentration. PMID- 15114236 TI - Comments on using the time of maximum effect site concentration to combine pharmacokinetics and pharmacodynamics. PMID- 15114238 TI - Moreno y Maiz: a missed rendezvous with local anesthesia. PMID- 15114239 TI - Coca leaf and local anesthesia. PMID- 15114241 TI - Cervical transforaminal blocks should not be attempted by anyone without extensive documented experience in fluoroscopically guided injections. PMID- 15114243 TI - A concerning direction. PMID- 15114244 TI - Ultrasound imaging of brachial plexus. PMID- 15114246 TI - Anesthetic preconditioning versus anesthetic treatment: effects on ischemic injury in isolated hearts. PMID- 15114248 TI - Near-miss accident during magnetic resonance imaging by a "flying sevoflurane vaporizer" due to ferromagnetism undetectable by handheld magnet. PMID- 15114249 TI - Safety of indocyanine green as a vital stain. PMID- 15114250 TI - Unanticipated vomiting and pulmonary aspiration at anesthesia induction in a formula-fed 4-month-old infant. PMID- 15114251 TI - Society of Neurosurgical Anesthesia and Critical Care annual meeting. San Francisco, California, October 10, 2003. PMID- 15114252 TI - NovoSeven for non-hemophilia hemostasis. PMID- 15114253 TI - New indications for modafinil (Provigil). PMID- 15114254 TI - Epinastine ophthalmic solution (Elestat). PMID- 15114255 TI - [Stem cells - biology and therapeutic application]. AB - Enormous hope is connected with stem cells with regard to cell therapy, and this has become one of the most dynamically developing areas of science at the moment. A stem cell has unlimited potential for self-renewal. It appears that it can be a source of in vitro differentiated progeny cells capable of repairing damaged tissue. These review provides information about the biological properties of embryonic stem cells, i.e. ESs (embryonic stem cells), EGs (embryonic germ cells), and ECs (embryonic carcinoma cells). Possible human embryonic stem cell applications are described, with consideration of the desired cell line and the signals involved in their differentiation. The information about adult stem cells present - hemopoietic stem cells and the cells residing in selected tissues and organs: endothelium, pancreas, liver, epithelium, and gastrointestinal tract. Methods of their identification using the cell surfaces are also presented: the possibilities of in vitro transdifferentation, the phenomenon of in vivo plasticity, as well as morphological and genetic properties. Some topics of cell therapy and its clinical application in diabetics amplification are included. PMID- 15114256 TI - [Nociceptin/orphanin FQ (N/OFQ)--the opioid, antiopioid or neuromodulator?]. AB - Nociceptin/orphanin FQ (N/OFQ) is a neuropeptide discovered in the middle of the 1990. It possesses an amino acid sequence very similar to those of endogenous opioid peptides (particularly dynorphin A). However, N/OFQ lacks the N-terminal tyrosine necessary for activation of mu-, kappa- and delta- opioid receptors and therefore does not bind to opioid receptors but to its own nociceptin receptor (NOP). Opioid peptides also do not bind to the NOP receptor. In spite of structural similarities, the pharmacological profile of N/OFQ is different from and, in many cases, opposite to that of the opioids. Intracerebroventricular injection of N/OFQ induces hyperalgesia and decreases the analgesic actions of opioids, but induces analgesia when given intrathecally. N/OFQ blocks the rewarding effects of morphine, ethanol, and psychostimulants such as cocaine and amphetamine, but given alone it does not have rewarding effect. N/OFQ is metabolized to shorter fragments, such as N/OFQ(1-13), N/OFQ(1-11), N/OFQ(1-7) and N/OFQ(1-6). These fragments show biological activity. The effects of N/OFQ include regulation of the release of numerous neurotransmitters and hormones, as NOP receptors are located presynaptically in different brain structures. The aim of this review was to present current opinion on the role of N/OFQ in nociception, reward and drug dependence. PMID- 15114257 TI - [Amiodarone and the thyroid gland]. AB - Amiodarone is an iodine-rich drug. Its chronic administration may lead to disturbances in thyroid hormone metabolism and/or overt gland dysfunction. It causes an increased in serum fT4, rT3, and TSH concentrations and a decreased serum level of fT3 without thyroid dysfunction. Amiodarone may induce thyrotoxicosis (AIT--Amiodarone-induced thyrotoxicosis) or hypothyroidism (AIH- Amiodarone-induced hypothyroidism) in some persons. AIT occurs more frequently in areas with low iodine intake. The excess iodine contributes to excessive thyroid hormone synthesis-type I AIT or may lead to thyroiditis and a destructive process of thyroid follicular cells, resulting in excess thyroid hormone release-type II AIT. The mixed form of AIT also occurs. Type I AIT should be treated with antithyroid drugs alone or in association with potassium perchlorate, type II AIT benefits from treatment with glucocorticoids, whereas the mixed form of AIT is most effectively treated with a combination of thionamides, potassium perchlorate, and glucocorticoids. AIT often requires thyroidectomy after restoration of euthyroidism or radioiodine therapy, provided that 24-h thyroid radioactive iodine uptake values permit. AIH prevails in areas with high dietary iodine intake. It requires a discontinuation of amiodarone therapy and thyroid hormone (levothyroxine) replacement. It can remit spontaneously. Amiodarone and L thyroxine therapy is also possible. Baseline thyroid function tests, thyroid antibodies, and imaging examinations such as thyroid ultrasound on initial evaluation and follow-ups every 6 months must be carefully monitored before starting amiodarone therapy. PMID- 15114258 TI - [Effect of methotrexate on the immune response in selected experimental models]. AB - Methotrexate (MTX) is one of the immunosuppressory compounds applied in the prophylaxis and treatment of several diseases, including: rheumatoid arthritis, systemic lupus erythematosus, psoriasis, graft-versus-host disease and, in combination with other drugs, neoplastic diseases. Studies in humans and in animal model, of clinical disease, have demonstrated that MTX diminishes the clinical symptoms of various immunological disorders. MTX, an antagonist of folic acid synthesis, causes apoptosis in activated cells, primarily in the G1 and S phases of the cell cycle. One of its actions, is inhibition of the synthesis or activity of several proinflammatory cytokines. At high doses, MTX is cytotoxic to hemopoietic cells; low doses, however, promote hemopoiesis. MTX also induces the differentiation of monocytic tumor cells, which may explain, in part, its therapeutic effects in the treatment of some disorders. The compound suppresses both cellular and humoral immune response and mitogen-induced lymphocyte proliferation. These effects are dose- and time-dependent. The strongest suppressory activity is exerted when MTX is applied 24 or 48 hours after immunization or mitogen stimulation. Administration of MTX in unfavorable conditions such as stress or other diseases, diminishes its tolerance and increases its toxicity. Side-effects of MTX may be ameliorated by application of pharmacological synthetic agents and plant extracts. In summary, MTX has been an effective agent in suppressing immunological disorders (for 50 years) and is still finding many applications. PMID- 15114259 TI - Expression and functional role of serine/threonine phosphatases in rat esophagus. AB - BACKGROUND: Smooth muscle contraction is due to phosphorylation, while relaxation results from dephosphorylation of regulatory proteins such as the light chains of myosin (MLC20). Dephosphorylation of MLC20 is catalyzed by protein phosphatases (PP) types 1 and 2A. While some data are available on the expression and functional importance of PP1 and PP2A in vascular smooth muscle preparations, little is known of the importance of PP for force regulation in the gastrointestinal tract. MATERIAL/METHODS: Here we address the expression and functional importance of PP1 and PP2A in the rat esophagus. We measured both the isometric force of contraction and PP activity and also performed Western and Northern blotting for PPs in the tunica muscularis mucosae (TMM) and the muscularis mucosae propria (MM). RESULTS: Cantharidin (0.01-100 microM) inhibited PP activity in homogenates from TMM (IC50=20 KM) in a concentration-dependent manner. The mRNAs of catalytic and regulatory subunits (PP1 alpha, PP1 beta, PP2A alpha and M110) were detectable at 1.8 kb, 3.2 kb, 2.0 kb and 5.7 kb in both TMM and MM. Western blot analysis revealed PP1 in TMM and MM. PP2A (protein) was more abundant in TMM than in TMM. Cantharidin increased muscular tone in isolated preparations of TMM (starting at 100 microM) to 164.6 (14.8 % of predrug force at 300 microM (the highest concentration studied). This increase in force of contraction was comparable to that of 10 microM carbachol. CONCLUSIONS: The results suggest that PP activity can play an important role in the regulation of esophageal tone. PMID- 15114260 TI - Autoantibodies against Cajal bodies in systemic lupus erythematosus. AB - BACKGROUND: Cajal bodies (CB) are distinct sub-nuclear domains rich in small nuclear ribonucleoprotein particles (snRNPs); they are involved in pre-mRNA processing. Lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibody production against different nuclear molecules, including those involved in pre-mRNA processing. The aim of the present investigation is to assess the presence of anti-CB autoantibodies in a cohort of SLE sera. MATERIAL/METHODS: Antinuclear antibodies (ANA) were screened by indirect immunofluorescence in a batch of 190 sera from patients who met the ACR criteria for SLE classification; fine specificity was determined by Western blot using HEp 2 cells or rat hepatocyte extracts purified by ion exchange chromatography. RESULTS: Four sera had anti-Cajal body (CB) autoantibodies. Interestingly, all of these patients had intermittent extensive oral and esophageal ulceration. The autoantibodies to CB were of the IgG class, and by Western blot these sera had reactivity against an 80 kDa protein (coilin) associated with Sm proteins. CONCLUSIONS: Anti-CB autoantibodies constitute an uncommon specificity of SLE; therefore it seems that anti-CB antibody specificity is associated with extensive mucous ulceration. PMID- 15114261 TI - Expression of aryl hydrocarbon receptor (AHR) and aryl hydrocarbon receptor nuclear translocator (ARNT) mRNA expression in human spermatozoa. AB - BACKGROUND: The objective of this study was to determine if human spermatozoa express AHR (dioxin receptor) and ARNT and if mRNA levels for these transcription factors correlate with sperm concentration, motility or morphology. MATERIAL/METHODS: This was a case-controlled cohort study in which AHR and ARNT mRNA levels in subjects with normal and abnormal semen analysis were compared. Semen analysis was performed by CASA. AHR/ARNT mRNA levels were determined by multiplex RT-PCR using Glyceryl aldehyde 3-phospho dehydrogenase (G3PDH) as an internal control. RESULTS: Human sperm expresses abundant amounts of AHR and ARNT mRNA. There were no differences in AHR and ARNT mRNA levels in subjects with normal and abnormal semen analysis. AHR and ARNT levels did not correlate with sperm concentration, morphology or motility. There was a trend (p=0.07) for lower AHR mRNA expression in sperm of men with severe oligospermia. There was a significant negative correlation (r=-0.42, p<0.05) between sperm AHR and ARNT mRNA. CONCLUSIONS: The presence of AHR in sperm provides a mechanism by which environmental dioxins, polycyclic aromatic hydrocarbons and polyhalogenated biphenyls could directly influence sperm function. PMID- 15114262 TI - Identifying colorectal metastases in liver biopsies: the novel CDX2 antibody is less specific than the cytokeratin 20+/7- phenotype. AB - BACKGROUND: Immunohistochemistry plays an important role in tracing the primary site in metastatic tumors of unknown origin. Therefore, determining the cytokeratin (CK) 20/CK7 pattern of metastases is one of the most helpful procedures as the CK20+/CK7- pattern is typical of colorectal adenocarcinomas. Expression of CDX2 protein is a new, highly specific and sensitive marker of the intestinal origin of adenocarcinomas. In the present study we compared the sensitivity and specificity of CDX2 expression and the CK20+/CK7- phenotype in predicting the colorectal origin of liver metastases. MATERIAL/METHODS: The study was carried out on a consecutive series of 125 core-needle biopsies of metastatic adenocarcinomas of the liver. Most of the patients were followed up to death, and primary tumor localization could be established in 102 cases by a combination of clinical, radiological, histological and, in some cases, autopsy data. All the needle biopsies were immunohistochemically stained for CK7, CK20 and CDX2. CDX2 expression (at 10% and 50% cut-off levels) and the CK20/CK7 pattern of the metastases were correlated to the primary site established. RESULTS: The CK20+/CK7- pattern showed a specificity of 98.7% in predicting colorectal primary localization, which was superior to that of CDX2 expression at both cut-off levels (90% and 95.3% respectively). The sensitivity of CDX2 expression in these circumstances was 84% at the 10% cut-off, somewhat higher than that of the CK20+/CK7- phenotype (79.5%), but lower at the 50% cut-off level (72.7%). CONCLUSIONS: The CK20+/CK7- immunophenotype is more specific in predicting the colorectal origin of liver metastasis than CDX2 expression. PMID- 15114263 TI - Serum cathepsin B activity during regression of Morris hepatoma 5123 D. AB - BACKGROUND: Serum cathepsin B activity has been considered a potential marker of tumor progression. We previously demonstrated that low-frequency electromagnetic stimulation (LF EMS), called bioresonance therapy (BRT), may both accelerate and inhibit the growth of transplantable hepatoma (Morris type 5123D), including total tumor regression. The aim of this study was to assess serum cathepsin activity during tumor progression and regression. MATERIAL/METHODS: Of 60 female rats inoculated with Morris hepatoma cells, 45 were treated with BRT, and the remaining 15 were left without treatment. Fifteen rats without inoculated tumors served as controls. Serum cathepsin B activity was determined, tumor volumes were measured, and histological examinations of the tumor tissues were performed. RESULTS: Of the 45 BRT-treated rats, tumor regression was observed in 31 rats, and serum cathepsin activity was analyzed in these rats. In all non-treated rats, tumor progression was observed. Serum cathepsin B activity was significantly higher in both the BRT-treated group (27.8+/-4.1 U/l, p<0.01) and the tumor bearing group (19.9+/-2.5 U/l, p<0.05), as compared to the controls (13.3+/-3.4 U/l). CONCLUSIONS: Cathepsin B may play an important role, not only in tumor expansion, but also during the processes of cancer cell death and resorption. High circulating levels may thus correspond to effective therapeutic response in the course of antitumor treatment. PMID- 15114264 TI - Immunohistochemical analysis of the interstitial mast cells in acute rejection of human renal allografts. AB - BACKGROUND: Morphometric investigations were performed to determine whether mast cells correlate with tubulointerstitial fibrosis in acute renal transplant rejection (ARTR) and to examine the relationship between mast cells, interstitial alpha-smooth muscle actin (alpha-SMA) expression, and interstitial infiltrates. MATERIAL/METHODS: Twenty-four renal allograft biopsy specimens from patients with ARTR were examined quantitatively by means of a computer image analysis system. As a control 11 allograft biopsy specimens were used from patients with no signs of rejection. RESULTS: The morphometric study revealed that the mean values of the interstitial tryptase positive cells, expression of alpha-SMA, interstitial volume, CD 68+, CD 43+ and CD 20+ cells were significantly increased in ARTR patients in comparison to controls. In the ARTR group there were significant positive correlations between interstitial tryptase positive cells and interstitial expression of alpha-SMA, interstitial volume, CD 43+ and CD 68+. Moreover, CD 68 positive cells significantly correlated with interstitial volume (r=0.51, p<0.02). The correlations between interstitial tryptase positive cells and CD 20+ cells as well as between CD 43+ and CD 20+ cells and interstitial volume were positive, but did not reach statistical significance. CONCLUSIONS: Our findings demonstrate that mast cells are one of the constitutive cell types in the interstitium in ARTR. Although the significant positive correlations between interstitial mast cell count and relative interstitial volume, as well as interstitial expression of alpha-SMA, suggest that these cells have a role in the development of early interstitial fibrosis in renal allografts, these relationships need further investigation. PMID- 15114265 TI - Effect of massage on blood flow and muscle fatigue following isometric lumbar exercise. AB - BACKGROUND: This study attempted to investigate the influence of massage on the skin and the intramuscular circulatory changes associated with localized muscle fatigue. MATERIAL/METHODS: Twenty-nine healthy male subjects participated in two experimental sessions (massage and rest conditions). Subjects lay prone on the table and were instructed to extend their trunks until the inferior portion of their rib cage no longer rested on the table. Subjects held this position for 90 seconds (Load I). Subjects then either received massage on the lumbar region or rested for 5 minutes, then repeated the same load (Load II). Skin blood flow (SBF), muscle blood volume (MBV), skin temperature (ST), and subjects' subjective feelings of fatigue were evaluated using Visual Analogue Scale (VAS). RESULTS: An increase of MBV between pre- and post-load II periods was higher after massage than after rest (p<0.05). An increase of SBF at pre- and post-load II was observed only under massage condition. An increase of SBF between post-load I and pre-load II periods was higher after massage than after rest (p<0.05). An increase of ST between post-load I and post-load II periods was greater after massage than after rest (p<0.05). The VAS score was lower with massage than with rest in the post-treatment period (p<0.01). CONCLUSIONS: A significant difference was observed between massage and rest condition on VAS for muscle fatigue. Lumbar massage administration also appeared to have some effect on increasing skin temperature and enhancement of blood flow in local regions. PMID- 15114266 TI - The effects on infants of potent antiretroviral therapy during pregnancy: a report from Spain. AB - BACKGROUND: The purpose of our study was to assess the effects on infants of protease inhibitor (PI)-based antiretroviral therapy (ART) given to their HIV positive mothers during pregnancy. MATERIAL/METHODS: A multicenter observational study was carried out at 11 centers in Spain, involving 124 HIV-1-infected pregnant women under ART and their infants. The mothers were classified according to the ART protocols used during pregnancy into two groups: group A, 52 women with > or =2 nucleoside reverse transcriptase inhibitors (NRTI) with or without NNRTI, for a mean time of 4.7+/-2.2 months; and group B, 72 women on protease inhibitor (PI)-based regimens for 5.4+/-2.6 months. RESULTS: Maternal therapy was well tolerated, with no serious adverse effects on pregnancy course. No newborn was infected with HIV-1. There were two deaths at birth (group B), both with extreme prematurity. Among the 126 ART-exposed infants (4 siblings), the most common toxicity was anemia (29%), without significant differences between the two groups. Low birthweight and prematurity were also common (21% and 14%, respectively). CONCLUSIONS: Optimal management of HIV-1 infection in women, regardless of their pregnancy status, can be recommended in more developed countries, without adverse effects on pregnancy outcome, and dramatically decreasing vertical transmission. HAART with PI versus potent ART during pregnancy was effective and safe for infants throughout the 12-month follow-up. In the light of recent advances in anti-HIV-1 pregnancy therapy, the long-term safety of these prophylactic and therapeutical strategies should be studied. PMID- 15114267 TI - Type 1 diabetes mellitus in Catalonia: chronic complications and metabolic control ten years after onset. AB - BACKGROUND: The purpose of this article is to describe the prevalence of microvascular (retinopathy, nephropathy and neuropathy) and acute diabetic complications, and the metabolic control status, in Catalonian patients with type 1 diabetes mellitus ten years after diagnosis. MATERIAL/METHODS: We performed a cross-sectional population study evaluating 427 Type 1 diabetic patients diagnosed between 1987 and 1988 in 15 hospitals in Catalonia. 278 subjects were located, and all the study parameters were collected from their hospital medical records. Mean age at onset was 13.8+/-6.9 years, and 56.5% were male. The mean age of the patients was 24.8+/-6.7 years. Albumin excretion rate (AER), plasma creatinine, lipid profile, glycosilated hemoglobin (HbA1c), blood pressure, presence of retinopathy and clinical polyneuropathy, and diabetes control were evaluated. RESULTS: The mean HbA1c was 7.8+/-1.7%. HbA1c was <7.5% in 48% of patients and >10% in 11.5%. The prevalence of retinopathy was 7.6%. An AER higher than 20 Kg/min was found in 10.4%. Neuropathy was present in 4.3%. A significant association between microvascular complications as a whole (retinopathy and/or microalbuminuria and/or clinical polyneuropathy) and HbA1c (p=0.04) and hypertension (p=0.04) was observed. There were no differences in diabetic complications regarding sex or age at presentation. CONCLUSIONS: This is one of the first studies in which the prevalence of diabetic microvascular complications is reported in a South European cohort of subjects registered in EURODIAB. This population-based study confirms that complications are already present ten years after diabetes onset in a non-negligible percentage of patients. PMID- 15114268 TI - Seasonal variation in the occurrence of deep vein thrombosis. AB - BACKGROUND: The occurrence of several adverse cardiovascular events, eg, myocardial infarction and stroke, is not randomly distributed over time, but shows definite seasonal patterns. The temporal pattern for pulmonary embolism (PE) is known, whereas the data concerning deep vein thrombosis (DVT) are not conclusive. The aim of our study was to investigate whether a seasonal variation exists also for DVT, together with the possible influence of different comorbid risk factors. MATERIAL/METHODS: We observed 1164 consecutive cases of DVT at the General Hospital of Ferrara, Italy, from 1998 to 2002. The total sample was divided into subgroups by gender, and by the most common comorbid conditions. For statistical analysis, the distribution of symptom onset was tested for uniformity by the chi 2 test for goodness of fit, and partial Fourier series were used to assess rhythmicity. RESULTS: Seasonal analysis showed a significantly reduced frequency of DVT events in summer and increased in winter, for the total population (p<0.0001), men (p=0.003), women (p=0.007), subjects with PE (p=0.001), and hypertension (p=0.001). Similarly, chronobiological analysis yielded a significant seasonal variation, with a peak in December for the total population (p<0.001), men and women (p<0.001 and p=0.02, respectively), PE (p=0.006), and hypertension (p=0.028). CONCLUSIONS: This study shows the existence of a highly significant seasonal pattern in the occurrence of DVT, characterized by a winter peak. Thus, colder months and relative hypercoagulability could be considered a supplementary high risk condition when prescribing an anticoagulant regimen. PMID- 15114269 TI - Diagnosis of migrainous vertigo: validity of a structured interview. AB - BACKGROUND: Migraine and vestibular symptoms are co-morbid. Migraine is increasingly recognized as a cause of benign, recurrent vertigo. Although the International Headache Society does not currently include the diagnosis of migrainous vertigo, specific criteria have been proposed and utilized in clinical trials. MATERIAL/METHODS: Seventeen adult migraineurs were separately screened for the diagnosis of migrainous vertigo using a standardized structured interview for migrainous vertigo (SIM-V) and a clinical evaluation by an experienced neurotologist. The SIM-V was administered by a nurse who read and recorded subject responses, obtaining no additional information for the diagnosis of migrainous vertigo. Comparison of diagnoses between clinician and SIM-V was made using Cohen's kappa reliability testing. Subjects were asked to return for a second interview by the same nurse using the SIM-V, at least 2 weeks after the initial assessment. Testing stability was evaluated by comparing diagnoses obtained with each SIM-V administration. RESULTS: Cohen's kappa demonstrated excellent test validity (kappa=0.75). Fourteen subjects returned for repeat testing, with excellent retest stability (kappa=0.85). The additional three subjects could not be relocated or were not interested in attending a retest appointment. CONCLUSIONS: The SIM-V is an easy-to-administer screening tool for the diagnosis of migrainous vertigo. Diagnostic comparison to a standard clinical assessment shows good test validity. Retest stability was also demonstrated. The SIM-V may be a useful screening tool for migraineurs with an additional complaint of dizziness. PMID- 15114270 TI - Duration of Raynaud's phenomenon is negatively correlated with serum levels of interleukin 10 (IL-10), soluble receptor of interleukin 2 (sIL2R), and sFas in systemic sclerosis patients. AB - BACKGROUND: Dysfunction of blood vessels and endothelial cells is regarded as a primary triggering factor of the development of autoimmunological processes in systemic sclerosis (scleroderma, SSc). Literature data suggest that the duration of Raynaud's phenomenon may correlate with disease severity. MATERIAL/METHODS: The study comprised 34 patients with SSc, 19 with limited SSc (lSSc) and 15 with diffuse SSc (dSSc). The duration of Raynaud's phenomenon (before skin fibrosis development) was correlated with the serum levels of selected markers, measured by ELISA, reflecting disturbances in autoimmunological processes, angiogenesis, apoptosis, and fibrosis. RESULTS: A shorter duration of Raynaud's phenomenon was shown to correlate with higher serum levels of interleukin-10 (IL-10) in the whole SSc group and of aminoterminal propeptide of collagen III (PIIINP) in the lSSc subgroup (p<0.05). It also correlated with higher serum levels of soluble interleukin 2 receptor (sIL2R) and the soluble protein sFas in the whole SSc group and the lSSc subgroup (p<0.05). CONCLUSIONS: The results obtained confirm the importance of vasomotor disturbance duration in triggering fibrosis in the course of systemic sclerosis. PMID- 15114271 TI - Incidence of renal insufficiency in cancer patients and evaluation of information available on the use of anticancer drugs in renally impaired patients. AB - BACKGROUND: Chronic renal failure is a common pathology. The high frequency of this disease in the general US population has been assessed in the NHANES III study. However, the frequency of chronic renal insufficiency among cancer patients remains unclear. MATERIAL/METHODS: 316 cancer patients were in a one month study, included regardless of their pathology, treatment (antineoplastic drugs used or programmed to be used, pretreated, or no treatment), or any other criteria. RESULTS: Among the patients, 287 (90.8%) had normal serum creatinine levels (<110 micromol/l), i.e. a frequency of 9.2% for renal insufficiency in our population. However, when renal function was estimated by calculating creatinine clearance using the Cockcroft and Gault formula, 33% of the patients had an estimated GFR of less than 80 ml/min. Among these, 28% had a creatinine clearance ranging form 80 to 50 ml/min and 5% had a creatinine clearance of less than 50 ml/min. Renal insufficiency is frequent in cancer patients since almost one third of the patients present renal insufficiency. Furthermore, among patients with normal serum creatinine levels, one patient out of five has asymptomatic renal insufficiency. CONCLUSIONS: Therefore, it is of major importance that renal function be assessed by calculation of creatinine clearance using the Cockcroft and Gault formula in every patient, even when serum creatinine is within the normal range. PMID- 15114272 TI - Conjunctival and tear film changes after vitamin C and E administration in non insulin dependent diabetes mellitus. AB - BACKGROUND: The purpose of our research was to investigate the effect of supplementation with vitamin C and E on ocular surface cytology specimens and related parameters in diabetic patients. MATERIAL/METHODS: 60 patients were enrolled in the study. The patients were given vitamin C (1000 mg/day) and vitamin E (400 IU/day) for 10 days. Conjunctival brush cytology specimens were obtained before and after treatment. Schirmer tests, break-up time and ocular ferning tests were also performed. RESULTS: Goblet cell densities were 50 cells/per field before and 59 cells/per field after supplementation (p=0.002). The stage of squamous metaplasia was 1.12+/-0.42 before and 0.88+/-0.41 after supplementation (p=0.011). The changes were accompanied with improved values for the Schirmer test (p<0.001), break up time (p=0.001), and ocular ferning (p<0.001). CONCLUSIONS: Diabetes mellitus is associated with increased oxidative stress. Our study suggests that supplementation with antioxidant vitamins C and E probably plays an important role in improving the ocular surface milieu. PMID- 15114273 TI - The evaluation of noise-induced hearing loss with distortion product otoacoustic emissions. AB - BACKGROUND: Distortion product otoacoustic emissions (DPOAEs) is an objective, sensitive test of cochlear function. The aim of this study was the evaluation of noise-induced hearing loss in a group of industrial workers, using this method in conjunction with standard pure-tone audiometry (PTA). MATERIAL/METHODS: Thirty four subjects (68 ears) were included in the study. Standard PTA, tympanometry, and DPOAEs were performed. DPOAEs were recorded in the format of the DP-gram, using an ILO Otodynamics analyzer (ILO292 DP Echoport) connected to a portable personal computer. Results were analyzed and compared with the data of 30 normal persons of similar age and sex, examined at the same laboratory, using identical stimulation and recording conditions. RESULTS: Most subjects had normal hearing thresholds for lower frequencies, but presented with a notched pattern in the 3000 to 6000 Hz region. The DPOAEs of the patients had reduced amplitudes compared with controls, or were even absent, in one or more frequencies. Across the frequency range of 1000 to 6000 Hz, a higher proportion of subjects showed abnormalities in DPOAEs than in the audiogram. CONCLUSIONS: From our data it may be concluded that cochlear dysfunction in noise-induced hearing loss may extend beyond the frequency region suggested by the audiogram, and that DPOAEs is a sensitive test of this damage. PMID- 15114274 TI - Gastric myoelectrical activity and intracranial hypertension. AB - BACKGROUND: The aim of the study was to evaluate altered patterns of gastric myoelectrical activity in patients with high intracranial pressure due to severe craniocerebral injury producing intracranial hematomas, or to neoplastic processes in the central nervous system. MATERIAL/METHODS: The study population consisted of 50 patients admitted to the Department of Neurotraumatology, Jagiellonian University College of Medicine. The controls were 16 healthy volunteers matched for gender and age. Gastric myoelectrical activity was recorded with standard cutaneous electrodes using the Synectics (Sweden) system of data storage and analysis. RESULTS: The most significant changes in gastric myoelectrical activity were observed in patients after severe head injury with increased intracranial pressure (Glasgow Coma Scale score 4-7). The percentage of bradygastria increased to 46.5 in these patients, and the signal amplitude was also increased. Significant dysrrhythmias occurred in patients with increased intracranial pressure due to brain tumours. The percentage of tachygastria increased to 36.5. CONCLUSIONS: The greatest changes in gastric myoelectrical activity were found in patients with increased ICP and coma (GCS score 4-7) due to head injury. Bradygastria was found in 46.5% of cases. PMID- 15114275 TI - Subclinical hypothyroidism in elderly women attending an outpatient clinic. AB - BACKGROUND: The object was to determine the frequency of subclinical hypothyroidism (SH) in elderly women and study its relation to serum lipids, hypertension, diabetes, and ischemic heart disease. MATERIAL/METHODS: A sample of 257 patients was randomly selected among women above the age of 50 visiting the King Abdulaziz University outpatient clinic. All were examined for thyroid function. Positive cases of SH were further tested for thyroid antibodies, hypothyroid symptoms, and goiter. Data were collected from the cases and a control group regarding age, presence of hypertension, diabetes, and ischemic heart disease, and lipoprotein levels and body mass index (BMI) values. Comparative analysis was performed between the cases and controls regarding dyslipidemia, BMI, hypertension, diabetes, and ischemic heart disease after age adjustment with logistic regression. RESULTS: Ninety patients out of 257 (35%) had SH. Positive thyroid antibodies were present in 55 (61%), goiter in 8 (9%), and hypothyroid symptoms in 22 (24%). In multiple regression analysis, SH cases were found to have lower risk for hypertension, ischemic heart disease, and diabetes than the controls after adjustment for age: OR 1.5 (95% CI: 0.8-2.83), 2.17 (0.75-6.28), 2.67 (1.50-4.76), respectively; the p-values were found to be significant for diabetes (0.19, 0.14, <0.001). However, there were no significant differences between cases and controls regarding measurements of LDL and cholesterol: 3.3+/-1.1 mmol/l and 5.5+/-1.2 mmol/l vs. 3.4+/-0.9 mmol/l and 5.4+/ 1.1 mmol/l, respectively. CONCLUSIONS: There is a high prevalence of SH among our sample, with no increased risk of hypertension, hyperlipidemia, ischemic heart disease, or diabetes. PMID- 15114276 TI - Perinatal supplementation of long-chain polyunsaturated fatty acids, immune response and adult diseases. AB - Both omega-6 and omega-3 long-chain polyunsaturated fatty acids (LCPUFAs) modulate TH1 and TH2 cell generation, their cytokine production, and cell proliferation and thus may serve as endogenous anti-inflammatory molecules. LCPUFAs suppress the production of tumor necrosis factor-alpha (TNF-alpha) (and so also of OX40, since it belongs to the family of TNFR) and the expression of Bcl-2, suggesting that these fatty acids have the ability to prevent/suppress autoimmune diseases. Human breast milk contains substantial amounts of both omega 3 and omega-6 fatty acids. This indicates that LCPUFAs present in human breast milk suppress the levels of OX40 and decrease the expression of Bcl-xL and Bcl-2 on exposure to self-antigens and thus, protects against the development of autoimmune diseases in later life. In view of this, I propose that supplementation of appropriate amounts of LCPUFAs during perinatal period protects against atopy, asthma, auto-immune diseases, type 1 and type 2 diabetes mellitus, hypertension, coronary heart disease, metabolic syndrome X, lymphomas, leukemias and other cancers, schizophrenia, depression and other adult diseases in which low-grade systemic inflammation plays a significant role. It is also likely that perinatal supplementation of LCPUFAs in adequate amounts modulates the expression of genes concerned with immune response, angiogenesis, central osmo/sodium and glucose sensors etc. This renders various tissues and organs including T cells and macrophages, endothelial cells, hypothalamic neurons, and various cardiovascular tissues to be able to counteract the pathological mechanisms that tend to induce various adult diseases by blunting the inflammatory responses in those who received adequate amounts of LCPUFAs during the perinatal period compared to those who did not. PMID- 15114277 TI - Open access publishing: a boon for scientific community. PMID- 15114278 TI - Analysis models to assess cost effectiveness of the four strategies for the work up of solitary pulmonary nodules. AB - BACKGROUND: To evaluate the role of computed tomography (CT) -guided needle biopsy and sodium iodide fluorine-18 fluorodeoxyglucose positron emission tomography (FDG-PET) in the investigation of solitary pulmonary nodules (SPN), which are discovered on screening chest radiographs, and to determine the cost effectiveness of these modalities. MATERIAL/METHODS: We have used decision-tree analysis models to assess the accuracy and cost-effectiveness of four strategies for the diagnosis and management of SPNs: CT alone strategy (baseline), CT plus FDG-PET strategy, CT plus FDG-PET plus CT-guided needle biopsy strategy, and CT plus CT-guided needle biopsy strategy. Reported values of prevalence of cancer, and sensitivity and specificity of each diagnostic modality were applied to the decision-tree models using Japanese health care costs. RESULTS: The prevalence of lung cancer among SPNs discovered on the lung cancer screening was less than 10%. In this prevalence, the strategies using CT-guided needle biopsy were the cost effective alternatives to the CT alone strategy (cost saving was 436,470 yen - 456,478 yen per patient), and had higher accuracies (95-96% vs. 67%). Both effects were mainly the result of reducing the number of the candidates who undergo unnecessary thoracotomy for a benign SPN, and these results were true over a wide range of prevalence of cancer (0-55%). The cost saving and accuracy of the CT plus FDG-PET strategy (359,206 and 92%) were approaching to those of the strategies using CT-guided needle biopsy. CONCLUSIONS: The introduction of CT guided needle biopsy and FDG-PET for the evaluation of SPNs, which are discovered on screening chest radiograph, is potentially cost-effective in Japan with high accuracy. PMID- 15114279 TI - Cost-effectiveness of positron emission tomography for non-small cell lung carcinoma in Canada. AB - BACKGROUND: Several studies over the past decade have demonstrated that 2-fluoro 2-D-[18F]fluorodeoxyglucose (FDG) positron emission tomography (PET) is more accurate than computed tomography (CT) for the staging of non-small cell lung carcinoma (NSCLC). This study uses quantitative decision tree modeling and sensitivity analysis to assess the cost-effectiveness of both a CT- and a CT+PET based management strategy for staging NSCLC in Canada. Both management costs and life expectancy are determined. MATERIAL/METHODS: Two patient management scenarios were compared--one using CT alone and one using both CT and PET. A survey of recent literature was used to construct a meta-analysis of available studies for the accuracies of CT and PET in staging NSCLC. Life expectancies were determined from recent Canadian statistics, and expected life expectancies with disease were calculated from knowledge of published survival rates. Management costs were determined from: estimates of the installation cost of PET facilities in Canada; management costs from our institutions; and recently published Canadian cost estimates of various procedures. RESULTS: A cost savings of 1455 Canadian dollars per person is expected for the CT+PET strategy, along with an increase in life expectancy (3.1 days), when compared with the CT alone strategy. This cost savings remained in favour of the CT+PET strategy when subjected to a rigorous sensitivity analysis. CONCLUSIONS: PET may be a cost effective means of staging NSCLC in Canada. PMID- 15114280 TI - Drug monitoring of pyrimethamine during maintenance therapy of toxoplasmic encephalitis in patients with advanced HIV infection during HAART. AB - BACKGROUND: Prophylaxis of toxoplasmic encephalitis was performed with pyrimethamine in 6 patients with advanced HIV infection during combination therapy with protease inhibitors. MATERIAL/METHODS: Steady-state plasma pyrimethamine (PYR) levels were measured by gas chromatography. Protease inhibitor plasma concentrations were analyzed trough concentration by high pressure liquid chromatography. During a treatment period of 22I13 months a total of 93 samples from 6 patients were investigated, containing pyrimethamine and protease inhibitors. RESULTS: The mean pyrimethamine concentration was 1,108+/ 459 ng/ml with a dosage of 37.5 mg/d and 1, 685+/-665 ng/ml with 50 mg/d. With the simultaneous use of indinavir (IDV), a mean pyrimethamine concentration of 2,165+/-273 ng/ml was found, significantly higher than in combination with saquinavir (SQV) and ritonavir (RTV) (1,192+/-178 ng/ml) or saquinavir and nelfinavir (NLV) (1,117+/-173 ng/ml). This effect was not considered clinically significant. Drug monitoring of protease inhibitors revealed a wide range of protease inhibitor levels. CONCLUSIONS: The statistically significant lower PYR concentrations with the SQV + RTV or SQV + NLV comedication in comparison to comedication with IDV were not considered clinically significant. Therapeutic drug monitoring of PYR and PI plasma levels can be recommended in patients during therapy with both PYR and PI, especially during therapy with a double PI regimen. PMID- 15114281 TI - Rifaximin plus mesalazine followed by mesalazine alone is highly effective in obtaining remission of symptomatic uncomplicated diverticular disease. AB - BACKGROUND: Rifaximin plus mesalazine has been showed to be more effective than rifaximin alone in the treatment of recurrent and complicated diverticulitis of the colon. We investigated the effectiveness of the combination rifaximin/mesalazine followed by mesalazine alone to evaluate tolerability and effectiveness in symptomatic remission in uncomplicated diverticular disease. MATERIAL/METHODS: We studied 90 consecutive patients (39 M, 51 F, mean age 67.2 yrs, range 32-91 yrs) with symptomatic uncomplicated diverticular disease. We assessed the following symptoms, scoring them on a quantitative scale: 1) constipation, 2) diarrhea, 3) abdominal pain, 4) rectal bleeding, and 5) mucus with stools. All were treated with 800 mg/day rifaximin plus 2.4 gr/day mesalazine for 10 days, followed by 1.6 gr/day mesalazine for 8 weeks. They were re-evaluated at the end of mesalazine-alone treatment. RESULTS: Eighty-six patients completed the study (95.56%): the total score decreased from 1439 to 44 (p<0.001). 70 patients (per-protocol: 81.40% (C.I.: 67-94%); on intention-to treat: 77.78% (C.I.: 60-85%)) were completely asymptomatic after the 8th week of treatment with mesalazine alone (total symptomatic score: 0), while 16 (per protocol: 18.60%; on intention-to-treat: 17.77%) showed only slight symptoms (total score: 44). Two (2.22%) showed recurrence of diverticulitis after 4 and 6 weeks of treatment with mesalazine alone. Two patients (2.22%) were withdrawn from the study for diarrhea after starting mesalazine. Two others (2.22%) showed transitory pruritus (one) and epigastric pain (one). CONCLUSIONS: The results show that rifaximin/mesalazine followed by mesalazine alone is extremely effective in resolving symptoms in patients with symptomatic uncomplicated diverticular disease. PMID- 15114282 TI - Olanzapine in the treatment of agitation in hospitalized patients with schizophrenia and schizoaffective and schizofreniform disorders. AB - BACKGROUND: Olanzapine is an atypical antipsychotic with proven therapeutic effect, though it is rarely used in agitated patients. This study was to assess the effectiveness and safety of a 20-mg initial dose of olanzapine to control agitation of hospitalized patients with schizophrenia and schizoaffective and schizofreniform disorders. MATERIAL/METHODS: A group of 95 patients aged 18-65 years was observed for 7 days. Effectiveness was assessed according to the CGI and PANSS (including the positive and negative symptoms and excitement subscale PANSS-EC) scales. Adverse events were also recorded. RESULTS: Two therapeutic groups were distinguished: patients treated with olanzapine alone (OLZ) and those whose condition necessitated at least one administration of benzodiazepines (OLZ + BZ). Although these groups differed with respect to baseline symptom severity, improvement was similar. In the OLZ group, improvement according to the CGI scale was observed after 2 h and persisted to the end of the study. In the OLZ + BZ group there was improvement after 1 h, but not after 2 h. Significant improvement reappeared after 6 h and persisted. Improvement measured by the PANSS-EC scale was noted in the OLZ group after 2 h and in the OLZ + BZ group after 6 h, persisting in both groups from the 12th hr to the end of the observation. CONCLUSIONS: Olanzapine is an antipsychotic which, used alone or in combination with benzodiazepines, is effective and well tolerated in treatment of agitation in schizophrenic patients. Further studies are warranted. PMID- 15114283 TI - The future of human embryonic stem cell research: addressing ethical conflict with responsible scientific research. AB - Embryonic stem (ES) cells have almost unlimited regenerative capacity and can potentially generate any body tissue. Hence they hold great promise for the cure of degenerative human diseases. But their derivation and the potential for misuse have raised a number of ethical issues. These ethical issues threaten to paralyze pubic funding for ES cell research, leaving experimentation in the hands of the private sector and precluding the public's ability to monitor practices, research alternatives, and effectively address the very ethical issues that are cause for concern in the first place. With new technology being inevitable, and the potential for abuse high, government must stay involved if the public is to play a role in shaping the direction of research. In this essay, I will define levels of ethical conflict that can be delineated by the anticipated advances in technology. From the urgent need to derive new ES cell lines with existing technology, to the most far-reaching goal of deriving genetically identical tissues from an adult patients cells, technology-specific ethical dilemmas can be defined and addressed. This staged approach provides a solid ethical framework for moving forward with ES cell research. Moreover, by anticipating the moral conflicts to come, one can predict the types of scientific advances that could overcome these conflicts, and appropriately direct federal funding toward these goals to offset potentially less responsible research directives that will inevitably go forward via private or foreign funding. PMID- 15114284 TI - Congenital scoliosis. AB - The management of congenital scoliosis requires a systematic approach with careful attention to detail. Any fortuitous diagnosis of vertebral anomalies in infancy, even if there is no significant scoliosis at that time on x-ray, requires frequent clinical and radiographic follow-up to detect progression. The presence of associated anomalies of the spinal cord, the kidneys and the heart should be evaluated by MRI, renal ultrasound or IVP, with cardiology evaluation as indicated. Curve progression or severe vertebral anomalies known to cause curve progression require immediate treatment to prevent deformity. Significant thoracic deformity, especially in a patient with thoracic insufficiency syndrome, is best treated with expansion thoracoplasty. The patient with congenital scoliosis requires a long term commitment to care with frequent orthopaedic follow-up throughout the growing years along with routine pulmonary function assessment once the patient is able to cooperate with testing. PMID- 15114285 TI - Challenge of personalized health care: to what extent is medicine already individualized and what are the future trends? AB - Personalized medicine: is it hype or revolution? In any case, there is not only real demand for it, but it also has a history. As it is, the personal aspects of health care have been partly neglected in the current era of evidence-based, scientific medicine. We now know that a 'one fits all' type of treatment has its limits. Medicine needs to be (re-)personalized. The time is right: the post genomic era provides the necessary molecular tools, but does it provide for the risks involved? Privacy, protection of minorities, and prevention of discrimination are at stake. Regulations are required. The health-care process needs redesigning to render personalized medicine effective. Information and communication management is challenged to handle the wealth of personal information and link to global medical knowledge. But the goal is magnificent: personal health planning, early diagnosis, the right drug for the right patient, and predictable side effects. PMID- 15114286 TI - 'Coalition-In-Iraq' countries 'internal' civil violent deaths compared to the USA 'external' violence of September 11th 2001. AB - BACKGROUND: The atrocity of 9/11 exemplified 'external' caused civil violent deaths, which have major policy implications for the countries contributing armed forces to the 'Coalition-In-Iraq' [C.I.I]. C.I.I Government's resources for perennial 'internal' violent deaths (suicide, homicide and road), are likely to be threatened by alternative priorities. C.I.I 'internal' deaths are compared with those of 9/11 to provide information to determine relative risks. MATERIAL/METHODS: To uniformly compare mortality between countries we utilise the most recent WHO mortality data, taking the latest three years 1997-1999 and calculating an average annual number and rate of 'internal' deaths in each country, these are compared with the 9/11 fatalities, calculating a proportional ratio, as an indicator of differential damage to families and society. RESULTS: USA 'external' deaths were 3,074 people and annual average suicides were 30,966. Total 'internal' deaths exceeded 'external' more than 30 times. Every fortnight there are more USA 'internal' violent deaths than on 9/11. Except Australia and Bulgaria, every country had more suicides than died on 9/11. Apart from Bulgaria, total 'internal' deaths exceeded the 'external' toll in all other CAI countries- by 14 times in Japan; 6 in Republic of Korea, 4 in Italy, 3 in Spain and the UK, twice in Canada and 1.5 times in Australia. CONCLUSIONS: The extent of 'internal' civil violent deaths, such as suicide, highlight the perennial pressures upon psychiatric services and the need to defend resources at a time of competing priorities. PMID- 15114290 TI - Hepatosplenic schistosomiasis: a model for the study of portal hypertension. PMID- 15114291 TI - Treatment of nonalcoholic fatty liver disease. AB - Treatment of patients with nonalcoholic fatty liver disease (NAFLD) has typically been focused on the management of associated conditions such as obesity, diabetes mellitus and hyperlipidemia. NAFLD associated with obesity may resolve with weight reduction, although the benefits of weight loss have been inconsistent. Appropriate control of glucose and lipid levels is always recommended, but not always effective in reversing the liver condition. Results of pilot studies evaluating ursodeoxycholic acid, gemfibrozil, betaine, N-acetylcysteine, vitamin E (alpha-tocopherol), metformin and thiazolidinedione derivatives suggest that these medications may be of potential benefit for patients with NAFLD. These medications, however, need first to be tested in well-controlled trials with clinically relevant end-points and extended follow up. A better understanding of the pathogenesis and natural history of NAFLD will help to identify the subset of patients at risk of progressing to advanced liver disease, and hence, those patients who should derive the most benefit from medical therapy. PMID- 15114292 TI - Intrahepatic cholestasis of pregnancy: changes in maternal-fetal bile acid balance and improvement by ursodeoxycholic acid. AB - Intrahepatic cholestasis of pregnancy (ICP) is a disease characterized by generalized pruritus and biochemical cholestasis that appears typically during the last trimester of gestation. The most predictive and accurate markers for diagnosis and follow-up of ICP are increased total bile acid levels (above 11,0 micromol/L), enhanced cholic acid percentage (above 42%) and decreased glycine/taurine bile acid ratio (below 1.0). Although essentially benign for the mother, evidence associates ICP with fetal poor prognosis resulting from increased transfer of bile acids from mother to fetus, who showed reduced ability to eliminate bile acids across the placenta. Those conditions lead to an accumulation of bile acids in the cord blood serum, meconium and amniotic fluid that may account for a diminished fetal well-being and sudden intra-uterine death by ICP. Ursodeoxycholic acid (UDCA) treatment was shown to reduce the bile acid content in the fetal compartment, while restoring the ability of the placenta to carry out vectorial transfer of these compounds towards the mother, decreasing bile acid levels in maternal serum and its passage to the fetus. In addition, UDCA administered to the mother also lowers the amount of bile acids present in colostrum without either increasing the UDCA concentration or causing major changes in lithocholic acid levels, further supporting the safety of UDCA in late pregnancy. Therefore, it is tempting to indicate UDCA as a first choice therapy for ICP as much as relevant aspects of fetal outcome may also be improved. This review focuses on the altered bile acid profiles in maternal and fetal compartments during ICP and its recovery by UDCA administration. Further elucidation of the precise mechanisms of action of UDCA and its therapeutic potential in improving fetal prognosis could result in the approval of UDCA for ICP treatment. PMID- 15114293 TI - Treatment of cirrhotic tense ascites with Dextran-40 versus albumin associated with large volume paracentesis: a randomized controlled trial. AB - Intravenous albumin infusion prevents complications after large-volume paracentesis (LVP), particularly paracentesis-induced circulatory dysfunction (PCD), and improves patient survival. However, albumin is expensive. We compared a low-molecular weight dextran (Dextran-40) with albumin in treating LVP in cirrhotic patients with tense ascites. Sixty-nine cirrhotic patients were included and 96 LVPs were performed. Any repeat punctures on the same patient were at least three months apart. Patients were randomized to receive either i.v. Dextran-40 infusion (Group I, n = 48) or i.v. albumin infusion after LVP (Group II, n = 48). Clinical, biochemical, and hormonal evaluations were done before and after LVP. Patients were followed up for the detection of any recurrence of ascites or complications. The two groups were similar in age, sex, and etiology of cirrhosis, and in the volumes of ascites recovered. Significant decreases in mean arterial pressure were observed in both groups 24 and 48 h after LVP. Urine volumes increased significantly at 24 h in both groups (p < 0.05), but remained high only in Group I. Plasma renin activity and aldosterone concentrations increased in both groups 48 h after LVP, but they were more marked in Group I. Complications developed in 17 % of patients treated with Dextran-40 and in 23 % treated with albumin (p > 0.05). Ascites recurrence rates and survival were similar in the two groups. In conclusion, Dextran-40 was thus not as efficacious as albumin for preventing PCD. PMID- 15114294 TI - Obesity-related non-alcoholic steatohepatitis and TGF-beta1 serum levels in relation to morbid obesity. AB - Non-alcoholic steatohepatitis (NASH) can vary from mild hepatic inflammation and steatosis to cirrhosis, and is most frequently associated with obesity, Type 2 diabetes mellitus, hypertension, and the female gender. The prevalence of fatty liver and NASH in the general population is 20% and 3%, respectively. In Western countries, 15-20% of the population is obese and 74-90% of them exhibit fatty changes in liver biopsies. We assessed the prevalence of NASH in morbidly obese patients and evaluated serum TGF-beta1 concentrations in different stages of liver fibrosis. Thirty-five obese patients were evaluated, nine male and 26 female. Their mean body mass index (BMI) was 43.62 +/- 7.92 kg/m2. Liver biopsies were evaluated by light microscopy; graded and staged according to Brunt's system. Serum obtained from patients was used to detect TGF-beta1 concentrations by an ELISA method. Serum alanine transaminase (ALT) levels were elevated in four of the patients and the mean level was 49.98 +/- 94.7 (8-65 IU/L). NASH was diagnosed in 32 (91%) of the biopsies, and the most common pattern seen was mixed, predominantly macrovesicular steatosis. Some degree of fibrosis was seen in 34 (97%) of the biopsies and 22 (63%) were at stage 2 (range 1-3). Serum concentrations of TGF-beta1 had no relationship with the stages of fibrosis. In conclusion, NASH and fibrosis are common in our obese patients, as observed in other studies. TGF-beta1 may play a key role in liver fibrogenesis. PMID- 15114295 TI - Zinc sulfate inhibits the enterohepatic cycling of unconjugated bilirubin in subjects with Gilbert's syndrome. AB - We have previously observed that UCB binds to ZnSO4 in vitro, and suppressed the biliary bilirubin secretion in the hamster. The aim of this study was designed to investigate whether Zn salts might inhibit the enterohepatic cycling of UCB in subjects with Gilbert s syndrome. Fifteen patients with Gilbert s syndrome and 5 normal healthy volunteers were included in this study according to the following criteria: fasting hyperbilirubinemia, no hemolysis, and free of any medication. Patients were randomly assigned to receive acute o chronic treatment. Subjects treated in acute form and normal healthy volunteers were treated with 40 mg of ZnSO4 in a single dose, where as patients treated in chronic form received 100 mg ZnSO4 in a single dose daily for 7 days. The serum UCB levels (mg/dL) decreased from 2.64 +/- 1.04 to 2.02 +/- 0.87 (p < 0.001) and 1.8 +/- 0.36 to 1.48 +/- 0.32 (p < 0.005) in subjects treated in acute an chronic form respectively, but not in the control group. Whereas, the serum Zn levels (mg/dL) increased from 96.3 +/- 16.8 to 118.8 +/- 19. 5, (p < 0.01) and from 117.6 +/- 8.5 to 130.7 +/- 6.6 (p < 0.03) in subjects treated in acute an chronic form and also in subjects in the control group (98.0 +/- 7.3 to 128.0 +/- 21.9) p < 0.03. This study showed that acute and chronic oral administration of ZnSO4 decreased serum UCB levels significantly in subjects with Gilbert s syndrome. Most likely by the inhibition of the "normal" enterohepatic cycling of UCB. PMID- 15114296 TI - Amebic liver abscess and liver cirrhosis. PMID- 15114297 TI - Amebic liver abscess. AB - Amebic liver abscess (ALA) highly endemic in most developing tropical countries is being encountered more frequently in other geographical areas maybe secondary to increased travel to areas where the disease is endemic as well as in the homosexual population. We report a classical clinical case of ALA in a 44 years old man diagnosed by ultrasound and positive seroameba titers who responded to oral imidazoles. PMID- 15114298 TI - Minimally invasive operation for breast cancer. PMID- 15114300 TI - Predictors of unsuccessful mechanical lithotripsy and endoscopic clearance of large bile duct stones. AB - BACKGROUND: Mechanical lithotripsy is used to break large bile duct stones. This study investigated the predictors of unsuccessful mechanical lithotripsy. METHODS: Consecutive patients with bile duct stones underwent endoscopic retrograde cholangiography, sphincterotomy, and basket removal of stones. Mechanical lithotripsy was performed for stones of large size (>15 mm diameter) that precluded extraction intact. Success was defined as complete clearance of the duct. Various predictive factors, including size and number of stones, stone impaction, serum bilirubin, presence of cholangitis, and bile duct diameter were analyzed in relation to the success or failure of lithotripsy. RESULTS: A total of 669 patients underwent endoscopic retrograde cholangiography for suspected choledocholithiasis, which was found in 401 patients. Of the latter patients, 87 had large stones that required mechanical lithotripsy. Lithotripsy was successful in 69 (79%) patients. Impaction of the stone(s) in the bile duct was the only significant factor that predicted failure of lithotripsy and consequent failure of bile duct clearance. Other factors, including stone size, were not significant. CONCLUSIONS: Mechanical lithotripsy is successful in about 79% of patients with large bile duct stones. The only significant factor that predicts failure of mechanical lithotripsy is stone impaction in the bile duct. PMID- 15114301 TI - Low-molecular-weight heparin does not prevent acute post-ERCP pancreatitis. AB - BACKGROUND: Studies suggest that heparin has anti-inflammatory effects that could prevent acute post-ERCP pancreatitis. The aim of this investigator-initiated, prospective, randomized, double-blind, multicenter study was to determine whether low-molecular-weight heparin can prevent acute post-ERCP pancreatitis. METHODS: Patients at increased risk for acute post-ERCP pancreatitis based on assessment of known risk factors were randomized to receive low-molecular-weight heparin (Certoparin 3000 IU subcutaneously) or placebo (saline solution 0.3 mL subcutaneously) the day before ERCP. The drug was given 2 hours before and 22 hours after ERCP. Documentation and follow-up included patient history, risk factors for acute post-ERCP pancreatitis, procedure-related data, assessment of pain (visual analogue scale, need for pain medication), laboratory findings before and after ERCP (0, 4, and 24 hours), as well as post-ERCP complications. The two-sided Fisher exact test was used for statistical comparison, and a p value < or =0.05 was considered significant. RESULTS: A total of 458 patients were enrolled in the study. Data from 10 patients could not be evaluated, leaving 221 patients in the low-molecular-weight heparin group and 227 in the placebo group (total 448 patients; 135 men, 313 women; mean age 58 [15] years). Low molecular-weight heparin and placebo groups were comparable with regard to risk factors for acute post-ERCP pancreatitis (gender distribution, age <65 years, history of pancreatitis, pancreas divisum, disorders of sphincter of Oddi) and procedure-related data (difficult cannulation, diagnostic or therapeutic ERCP, needle-knife papillotomy, endoscopic sphincterotomy, biliary or pancreatic procedure, pancreatic contrast injection, success and final diagnosis of ERCP). Acute post-ERCP pancreatitis occurred in 8.5% (38/448), with one death resulting from severe pancreatitis. Low-molecular-weight heparin offered no benefit compared with placebo based on the frequency of acute post-ERCP pancreatitis (low molecular-weight heparin, 18/221 vs. placebo, 20/227; p=0.87) and the severity of acute post-ERCP pancreatitis (low-molecular-weight heparin, 14 mild, 3 moderate, one severe; placebo, 18 mild, two moderate, 0 severe). The 24-hour serum amylase values and 24-hour pain scores did not differ significantly between the low molecular-weight heparin group and the placebo group. Bleeding complications occurred in two patients, both in the low-molecular-weight heparin group (one mild, one moderate). CONCLUSIONS: Prophylactic subcutaneous administration of low molecular-weight heparin does not prevent acute post-ERCP pancreatitis. PMID- 15114302 TI - Endoscopic papillary balloon dilation and endoscopic sphincterotomy for bile duct stones: long-term outcomes in a prospective randomized controlled trial. AB - BACKGROUND: Little is known about the long-term outcome of endoscopic papillary balloon dilation for removal of bile duct stones. A randomized trial that compared long-term outcomes after endoscopic papillary balloon dilation and endoscopic sphincterotomy was conducted. METHODS: Thirty-two patients with bile duct stones were randomized to endoscopic papillary balloon dilation or endoscopic sphincterotomy, with 16 patients in each group. Endoscopic papillary balloon dilation was performed by using an 8-mm-diameter balloon; endoscopic sphincterotomy was performed in the standard manner. The success rates for stone removal, as well as the frequency and types of early (<15 days), mid-term (<1 year), and long-term (1-6 years) post-procedure complications were evaluated. RESULTS: The success rates for stone removal and early complication rates were similar for both groups. The frequency of stone recurrence was approximately 4 fold higher in the endoscopic papillary balloon dilation group (25%) vs. the endoscopic sphincterotomy group (6.3%) at mid-term evaluation. However, over the long term, Kaplan-Meier estimated probability of stone recurrence tended to be higher in the endoscopic sphincterotomy group vs. the endoscopic papillary balloon dilation group; recurrent stones were found in, respectively, 26.7% vs. 6.3%. Complications occurred in 7 patients in each group. CONCLUSIONS: Long-term outcome of endoscopic papillary balloon dilation for bile duct stone removal is satisfactory, provided that consideration is given to recurrence of stones by early follow-up evaluation. PMID- 15114303 TI - Usefulness of three-dimensional, multidetector row CT (virtual gastroscopy and multiplanar reconstruction) in the evaluation of gastric cancer: a comparison with conventional endoscopy, EUS, and histopathology. AB - BACKGROUND: The aim of this study was to assess the role of three-dimensional multidetector row CT for detection, precise localization, and staging of gastric cancer by comparison with conventional endoscopy, EUS, and histopathology. METHODS: Sixty-three patients with gastric cancer (31 early stage, 32 advanced) were evaluated by EGD with biopsies, EUS, and three-dimensional multidetector row CT between January 2003 to August 2003. Three-dimensional multidetector row CT findings were analyzed by a single radiologist blinded to the endoscopic findings. Among 63 patients, the findings were confirmed in 48 at surgery or by EMR. In the remaining cases, the findings were confirmed by EGD and biopsy specimen. The accuracy of three-dimensional multidetector row CT for detection, localization, and staging of gastric cancer was determined, compared with endoscopy, EUS, and histopathology. RESULTS: Among the 63 patients, there were 67 gastric cancers. The overall accuracy of three-dimensional multidetector row CT for detection of gastric lesions was 94% (63/67), with accuracies of 96.7% (30/31) and 100% (32/32) for detection of, respectively, early and advanced stage gastric cancer. The overall accuracy, sensitivity, and specificity for EUS and three-dimensional multidetector row CT in the pre-operative determination of depth of invasion (T stage) were, respectively, 87.5%, 82.4%, and 96%; and 83.3%, 69.1%, and 94.4%. The accuracy, sensitivity, and specificity of EUS and three dimensional multidetector row CT for lymph node staging was, respectively, 79.1%, 57%, and 89.5%; and 75%, 57.4%, and 89.3%. CONCLUSIONS: Three-dimensional multidetector row CT, along with virtual gastroscopy is a promising method for pre-operative evaluation of gastric cancer. PMID- 15114304 TI - EUS-guided FNA of the left adrenal gland in patients with thoracic or GI malignancies. AB - BACKGROUND: The diagnostic yield and safety of trans-gastric EUS-guided FNA of the left adrenal gland are not well defined. METHODS: All patients with an enlarged left adrenal gland on abdominal imaging and known or suspected malignancy referred to two EUS centers over a 3-year period were included in this study. EUS-guided FNA was performed on an outpatient basis by one of 4 experienced endosonographers. RESULTS: Thirty-one consecutive patients (21 men, 10 women; mean age 64.8 years) were evaluated. Tissue adequate for interpretation was obtained in all patients; no attempt to obtain tissue was unsuccessful. The median number of needle passes was 4.5 (range 1-8). No immediate complications were encountered. EUS-guided FNA confirmed malignant left adrenal involvement in 42% (13/31) of the patients. Patients with malignant left adrenal masses were more likely to have known cancer at another site (OR 12.0: 95% CI[1.6, 87.9]). Patients with benign masses were more likely to have preservation of the normal sonographic appearance of the adrenal gland ("seagull" configuration) compared with those with malignant masses (OR 9.8: 95% CI[1.9, 51.0]). The accuracy of EUS imaging based on size (> or =3 cm) alone was 81%: 95% CI[63, 93]). Of the patients with malignant adrenal masses, 85% (11/13) died or their clinical condition deteriorated during follow-up, while 15% (2/13) were being treated and were stable clinically. CONCLUSIONS: EUS-guided FNA of the left adrenal gland is a minimally invasive, safe, and highly accurate method that confirms or excludes malignant adrenal involvement in patients with thoracic or GI malignancies. PMID- 15114305 TI - Diagnostic value of distal colonic polyps for prediction of advanced proximal neoplasia in an average-risk population undergoing screening colonoscopy. AB - BACKGROUND: For colorectal cancer screening, the predictive value of distal findings in the ascertainment of proximal lesions is not fully established. The aims of this study were to assess distal findings as predictors of advanced proximal neoplasia and to compare the predictive value of endoscopy alone vs. combined endoscopic and histopathologic data. METHODS: Primary colonoscopy screening was performed in 2210 consecutive, average-risk adults. Age, gender, endoscopic (size, number of polyps), and histopathologic distal findings were used as potential predictors of advanced proximal neoplasms (i.e., any adenoma > or =1 cm in size, and/or with villous histology, and/or with severe dysplasia or invasive cancer). Polyps were defined as distal if located in the descending colon, the sigmoid colon, or the rectum. Those in other locations were designated proximal. RESULTS: Neoplastic lesions, including 11 invasive cancers, were found in 617 (27.9%) patients. Advanced proximal neoplasms without any distal adenoma were present in 1.3% of patients. Of the advanced proximal lesions, 39% were not associated with any distal polyp. Older age, male gender, and distal adenoma were independent predictors of advanced proximal neoplasms. The predictive ability of a model with endoscopic data alone did not improve after inclusion of histopathologic data. In multivariate logistic regression analysis, the predictive ability of models that use age, gender, and any combination of distal findings was relatively low. The proportion of advanced proximal neoplasms identified if any distal polyp was an indication for colonoscopy was only 62%. CONCLUSIONS: A strategy in which colonoscopy is performed solely in patients with distal colonic findings is not effective screening for the detection of advanced proximal neoplasms in an average-risk population. PMID- 15114306 TI - A prospective evaluation of high-magnification chromoscopic colonoscopy in predicting completeness of EMR. AB - BACKGROUND: EMR is used to treat flat and sessile lesions in the colon. The aim of this study was to prospectively assess the efficacy of high-magnification chromoscopic colonoscopy in predicting complete resection margins after EMR. METHODS: A total of 1250 patients underwent colonoscopy by using a magnifying colonoscope. Chromoscopy with indigo carmine and crystal violet dye solutions was used to assess mucosal pit patterns. EMR was performed by using the saline solution inject-and-cut technique. After EMR, resection margins were inspected by using high magnification, and completeness of excision was predicted from the surface pit pattern. This was compared with completeness of excision as determined histopathologically. RESULTS: A total of 684 lesions were treated by EMR (62 piecemeal) in 602 patients. The sensitivity of high-magnification chromoscopic colonoscopy for predicting remnant tissue in the lateral margins and the deep margins were, respectively, 79% and 80%. Specificity for both margins was 97%. The overall accuracy of high-magnification chromoscopic colonoscopy in predicting incomplete resection after EMR in the lateral axis and the deep axis was, respectively, 93% and 95%, where the true respective fractions of incomplete resections were 17% and 10%. Of the single en bloc EMRs performed, 77 (12%) had histopathologic evidence of incomplete resection in either axis, compared with 60 (97%) of the 62 piecemeal resections. Piecemeal resection was more likely to result in incomplete resection compared with en bloc EMR (p < 0.001). Complete resection by EMR was more likely if the lesion was sessile instead of flat (p < 0.001). CONCLUSIONS: High-magnification chromoscopic colonoscopy as an in vivo modality for prediction of remnant tissue after EMR has a high overall accuracy, but further studies assessing long-term outcome and cost-effectiveness compared with conventional colonoscopic techniques are required. PMID- 15114307 TI - A prospective, observational study of colonic mucosal abnormalities associated with orally administered sodium phosphate for colon cleansing before colonoscopy. AB - BACKGROUND: Colonic mucosal abnormalities associated with sodium phosphate colonic preparations have been described. The aim of this prospective study was to determine the frequency at which these occur and to define the endoscopic and histopathologic patterns of these changes. METHODS: Within a 16-month period, 730 of patients referred for elective colonoscopy entered the study. Patients with known inflammatory bowel disease and those taking non-steroidal anti-inflammatory drugs were excluded. A sodium phosphate solution was ingested orally 14 and 8 hours before endoscopy. After standard colonoscopy, a 3-year clinical follow-up program was conducted. RESULTS: Endoscopically, mucosal lesions, possibly associated with sodium phosphate ingestion, were visible in 24 patients (3.3%). Erosions were found in 3 patients, aphthoid lesions in 21 patients, and an ulcer in one patient. Lesions often were multiple. Histopathologically, findings included focal active inflammation in 14 of 24 patients, mucosal disruption and erosion (7/24), edema of the lamina propria (5/24), mucosal hyperemia or focal hemorrhage (5/24), lymphoid nodules (5/24), and ulceration (1/24). CONCLUSIONS: Orally administered sodium phosphate-associated colonic mucosal abnormalities are infrequent but can mimic an non-steroidal anti-inflammatory drug-induced injury or inflammatory bowel disease, and in particular must be differentiated from Crohn's disease. PMID- 15114308 TI - Inter- and intra-observer variability in the measurement of length at endoscopy: Implications for the measurement of Barrett's esophagus. AB - BACKGROUND: Endoscopic measurements are frequently used to determine the length of Barrett's epithelium. Decreases in measured length also are used to assess response to pharmacotherapy. The aim of this study was to determine the accuracy of the endoscopic measurement of length under standardized conditions. METHODS: A life-size model of the esophagus was built by using an opaque polyvinyl tube. Transparent plastic inserts designed to resemble Barrett's epithelium were created. Ten different insert lengths were randomly studied. Trained endoscopists and fellows in training blinded to the lengths of the inserts performed measurements twice in random order. RESULTS: A total of 240 measurements were made by 12 endoscopists. The mean difference (1 standard deviation) between the measured and correct length was 1.10 (1.7) cm. Overestimation of the length occurred in 47% and underestimation in 37%. The kappa statistic for the first and second measurement (within 0.5 cm) performed by the same individual (intra observer) was 0.40, suggesting only fair agreement. There was considerable variability between endoscopists. CONCLUSIONS: Accurate measurement of length at endoscopy is difficult even under ideal conditions. Intra-observer agreement is fair, but results obtained by different endoscopists are widely divergent. Small improvements in the length of Barrett's epithelium observed in some clinical trials could be because of chance instead of therapeutic effect. PMID- 15114309 TI - Safety and effectiveness of ketamine as a sedative agent for pediatric GI endoscopy. AB - BACKGROUND: The ideal sedation for children undergoing GI endoscopy remains elusive. After ketamine was introduced as a sedative agent in our GI procedure suite, improved sedation and reduced complications were observed. The aim of this study was to assess the safety and effectiveness of ketamine as a sedative agent for GI endoscopy in pediatric patients. METHODS: A retrospective cohort study of 402 procedures (EGD, colonoscopy) was performed. Sedation-related complications were defined as hypoxia (oxygen saturation <95% by pulse oximetry), agitation, emergence reactions, stridor, laryngospasm, nausea, vomiting, aspiration, and muscle twitching, or any combination thereof. Sedation groups were defined as the following: Group I, midazolam and meperidine (n=192); Group II, midazolam, meperidine, and ketamine (n=82); and Group III, midazolam and ketamine (n=128). RESULTS: Group 1 (midazolam and meperidine) had the highest frequency of complications, most commonly hypoxia. Group 3 (midazolam and ketamine) had the lowest rate of complications (p=0.001) and the highest rate of adequate sedation, although the difference was not significant (p=0.07). CONCLUSIONS: The combination of midazolam and ketamine appears to provide safe and effective sedation for pediatric patients undergoing endoscopy. PMID- 15114310 TI - Biopsy specimen acquisition in patients with newly diagnosed peptic ulcer disease as determined from a national endoscopic database. AB - BACKGROUND: Eradication of Helicobacter pylori infection decreases peptic ulcer recurrence. Therefore, assessment of Helicobacter pylori status is recommended for patients with newly diagnosed peptic ulcer disease. METHODS: Data obtained from the Clinical Outcomes Research Initiative's national endoscopic database were analyzed to characterize the acquisition of biopsy specimens in patients with a non-bleeding gastric or duodenal ulcer newly diagnosed by EGD. RESULTS: Between January 2000 and June 2003, 8299 patients underwent EGD with identification of non-bleeding peptic ulcer disease in the stomach (5390) or the duodenum (2909). Overall, biopsy specimens were obtained from the gastric or duodenal ulcer in 5578 (67%) of these patients. Multivariate analysis identified male gender (odds ratio [OR] 0.75, 95% confidence interval (CI) [0.66-0.85] vs. female), age greater than 75 years (OR 0.67, 95% CI [0.57-0.77] vs. age <55 years), ulcer location (OR 0.53, 95% CI [0.48-0.59] for duodenal vs. gastric ulcers) and endoscopy setting (OR 0.35, 95% CI [0.31-0.39] for academic vs. community; OR 0.36, 95% CI [0.32-0.41] for Veterans Affairs medical centers vs. community) as independent predictors for the acquisition of biopsy specimens (p < 0.001 for all). CONCLUSIONS: The findings of this study suggest that there is variation in the rates of biopsy specimen acquisition among patients with ulcers who may be at risk for Helicobacter pylori infection. Given the established benefit of Helicobacter pylori eradication, further study is needed to determine whether physicians are diagnosing and treating Helicobacter pylori infection adequately in patients with peptic ulcer. PMID- 15114311 TI - Sphincter of Oddi dysfunction, part 2: Evidence-based review of the presentations, with "objective" pancreatic findings (types I and II) and of presumptive type III. PMID- 15114312 TI - Mechanical lithotripsy for large bile duct stones. PMID- 15114313 TI - Esophageal intramural pseudodiverticulosis. PMID- 15114314 TI - Cytomegalovirus-associated gastritis. PMID- 15114315 TI - Metastatic duodenal carcinoid tumor. PMID- 15114316 TI - Hemosuccus pancreaticus. PMID- 15114317 TI - Bronchial fistula. PMID- 15114318 TI - Right groin abscess. PMID- 15114319 TI - Evaluation of pancreatic cystic lesions with EUS. PMID- 15114320 TI - A novel technique for endoscopic sphincterotomy when using a percutaneous transhepatic cholangioscope in patients with an endoscopically inaccessible papilla. AB - BACKGROUND: Endoscopic sphincterotomy is difficult and sometimes impossible in patients who have undergone gastrectomy or partial gastrectomy with Billroth II reconstruction. For such patients, a novel technique was developed in which endoscopic sphincterotomy is performed via percutaneous transhepatic cholangioscopy. This report describes an initial experience with this technique. METHODS: After dilation of the percutaneous fistula, a cholangioscope with a push type sphincterotome attached was inserted into the bile duct via the fistula and then through the papilla into the duodenum. The tip of the instrument then was retroverted to obtain a frontal view of the papilla. Then, a sphincterotomy incision was extended to the proximal, orad margin of the papillary eminence. OBSERVATIONS: Percutaneous transhepatic biliary drainage was performed in 3 patients with obstructive jaundice and bile duct stones. In all patients, percutaneous transhepatic cholangioscopic sphincterotomy was performed successfully, without procedure-related complication. Thereafter, all stones and stone fragments cleared from the duct by spontaneous migration. CONCLUSIONS: Endoscopic sphincterotomy via percutaneous transhepatic cholangioscopy potentially is an innovative technique for endoscopic sphincterotomy in patients with an endoscopically inaccessible papilla. PMID- 15114321 TI - Endoscopic hemostatic treatment under irrigation for upper-GI hemorrhage: a comparison of one third and total circumference transparent end hoods. AB - BACKGROUND: Endoscopic hemostasis for upper-GI hemorrhage often is difficult to achieve if the view of the bleeding lesion is poor because of the presence of mucus, blood, and clots. An end hood that facilitates endoscopic hemostatic procedures while simultaneously allowing irrigation of the bleeding site was designed by us. Based on this design, a one-third partial irrigating end hood was developed, and its usefulness for treatment of non-variceal hemorrhage was evaluated. METHODS: The end hood was fabricated by drilling a side hole in the cap portion of a transparent end hood. An irrigation tube was glued to the exterior surface over the hole. A "total" (type 1) and a "one-third partial" (type 2) transparent end hood were fabricated. These differ with respect to the proportion of the endoscope circumference that is hooded by the device. The fabricated transparent end hood was placed on the tip of a standard endoscope. With the end hood in place, endoscopic hemostatic treatment under irrigation was performed in 35 patients (type 1 end hood, 18; type 2, 17) with non-variceal upper-GI hemorrhage. OBSERVATIONS: Hemostatic treatment was enhanced by simultaneous irrigation beneath the end hood, and hemostasis was successfully achieved in 34 of 35 cases. The time required to achieve hemostasis was significantly shorter in the type 2 group than the type 1 group (median 11.8 vs. 16.9 minutes; p < 0.05). CONCLUSIONS: The end hood was extremely useful for endoscopic hemostatic treatment under irrigation. The "one-third partial" end hood is superior to the total end hood in terms of duration of time required to achieve hemostasis. PMID- 15114322 TI - A novel endoscopic device for retrieval of polyps resected from the colon and rectum. AB - BACKGROUND: Retrieval of polyps resected from the colorectum is time consuming, despite the availability of various devices, such as grasping forceps, nets, and suction traps, for this purpose. A novel device for retrieval of polyps was developed and tested. METHODS: Patients with polyp(s) less than or equal to 6 mm in diameter were included in this study. The effectiveness of the device for polyp retrieval was evaluated. OBSERVATIONS: In total, 180 of 184 polyps were retrieved from 100 patients. Four polyp specimens (2.2%) were lost after polypectomy. None of the retrieved specimens were damaged. All retrieved polyps could be assessed histopathologically, and a definitive diagnosis was made. CONCLUSIONS: The newly developed device proved to be reliable when used in clinical practice. PMID- 15114323 TI - Complications of temporary pancreatic stent insertion for pancreaticojejunal anastomosis during pancreaticoduodenectomy. AB - BACKGROUND: Morbidity associated with pancreaticoduodenectomy usually results from complications associated with the pancreaticojejunal anastomosis, in particular, a pancreatic leak. Four patients with retained transanastomotic pancreatic stent-induced complications after pancreaticduodenectomy were identified. METHODS: Medical records for the 4 patients were reviewed, and telephone interviews were conducted. OBSERVATIONS: Each patient underwent pancreaticoduodenectomy for a peripapillary tumor with creation of a pancreaticojejunal anastomosis by using an internal 8F Silastic stent. Subsequent evaluation for steatorrhea (n=3) or recurrent pancreatitis (n=1) led to discovery of a retained pancreatic stent. In one patient, the stent was incidentally discovered. Steatorrhea significantly improved (n=1) or resolved (n=2) after stent removal. The patient with pancreatitis has not experienced another episode. CONCLUSIONS: The possibility of a retained stent should be considered in patients presenting with steatorrhea or pancreatitis after pancreaticoduodenectomy. PMID- 15114324 TI - ERCP by using a prototype oblique-viewing endoscope in patients with surgically altered anatomy. PMID- 15114325 TI - Heparin provocation for identification and treatment of a gastric Dieulafoy's lesion. PMID- 15114326 TI - Adenoma and early stage adenocarcinoma of the appendix: diagnosis by colonoscopy. PMID- 15114327 TI - Metastatic breast cancer to the gastroesophageal junction 14 years after radical mastectomy. PMID- 15114328 TI - Gastric hypothermic injury caused by accidental ingestion of dry ice: endoscopic features. PMID- 15114329 TI - Strongyloides colitis in a patient with Cushing's syndrome. PMID- 15114330 TI - Cytomegalovirus esophagitis with massive upper-GI hemorrhage. PMID- 15114331 TI - Endoscopic electrosurgical snare resection of leiomyosarcoma of main duodenal papilla. PMID- 15114332 TI - Cystic dilatation of the intraduodenal portion of the duct of Wirsung (Wirsungocele). PMID- 15114333 TI - Dieulafoy's lesion in a Meckel's diverticulum. PMID- 15114335 TI - Don't catch me if you can. PMID- 15114336 TI - A fine balance for life and death decisions. PMID- 15114337 TI - Catching the common cold. PMID- 15114338 TI - The dynamic tunnel. PMID- 15114340 TI - Structure and mechanism of the RNA polymerase II transcription machinery. AB - Advances in structure determination of the bacterial and eukaryotic transcription machinery have led to a marked increase in the understanding of the mechanism of transcription. Models for the specific assembly of the RNA polymerase II transcription machinery at a promoter, conformational changes that occur during initiation of transcription, and the mechanism of initiation are discussed in light of recent developments. PMID- 15114341 TI - Inhibition of the phosphodiesterase 4 (PDE4) enzyme reverses memory deficits produced by infusion of the MEK inhibitor U0126 into the CA1 subregion of the rat hippocampus. AB - Cyclic AMP-specific phosphodiesterase 4 (PDE4), which is an integral component of NMDA receptor-mediated cAMP signaling, is involved in the mediation of memory processes. Given that NMDA receptors also mediate MEK/mitogen-activated protein kinase (MAPK, ERK) signaling, which is involved in synaptic plasticity, and that some PDE4 subtypes are phosphorylated and regulated by ERK, it was of interest to determine if PDE4 is involved in MEK/ERK signaling-mediated memory. It was found that rolipram, a PDE4-selective inhibitor, reversed the amnesic effect in the radial-arm maze test of the MEK inhibitor U0126 administered into the CA1 subregion of the rat hippocampus. Consistent with this, rolipram, either by peripheral administration or direct intra-CA1 infusion, enhanced the retrieval of long-term memory impaired by intra-CA1 infusion of U0126 using the step-through inhibitory avoidance test. The same dose of rolipram did not affect U0126-induced reduction of phospho-ERK1/2 levels in the CA1 subregion. However, in primary cultures of rat cerebral cortical neurons, pretreatment with U0126 increased PDE4 activity; this was correlated with the U0126-induced reduction of phospho-ERK1/2 levels. These results suggest that MEK/ERK signaling plays an inhibitory role in regulating PDE4 activity in the brain; this may be a novel mechanism by which MEK/ERK signaling mediates memory. PDE4 is likely to be an important link between the cAMP/PKA and MEK/ERK signaling pathways in the mediation of memory. PMID- 15114342 TI - D-amphetamine boosts language learning independent of its cardiovascular and motor arousing effects. AB - D-Amphetamine (AMPH) was effective in a number of studies on motor and language recovery after stroke, but given safety concerns, its general use after stroke is still debated. Most stroke patients are excluded from treatment because of a significant risk of cardiovascular dysregulation. AMPH acts on multiple transmitter systems, and mainly the noradrenergic actions are related to the cardiovascular effects. If AMPH's cardiovascular and arousal effects were correlated with its plasticity-enhancing effects in humans, this would imply that desired and undesired effects are inevitably tied. If not, improved cerebral reorganization may not be mediated by AMPH's arousing effects and could be achieved with substances lacking the undesired cardiovascular effects. As a model for language recovery after stroke, we used a prospective, randomized, double blind, placebo-controlled design and taught 40 healthy male subjects an artificial vocabulary of 50 concrete nouns over the course of five consecutive training days (high-frequency training). The associative learning principle involved higher co-occurrences of 'correct' picture-pseudoword pairings as compared to 'incorrect' pairings. Subjects received either AMPH (0.25 mg/kg) or placebo 90 min prior to training on each day. Novel word learning was significantly faster and better in the AMPH as compared to the placebo group. Increased learning success was maintained 1 month post-training. No correlation was found between training success and drug-induced increases in blood pressure, heart rate, or a facilitation of simple motor reaction time. Our data show that AMPH's plasticity-enhancing effect in humans is not related to its cardiovascular arousal. This suggests that the beneficial effects in stroke patients could also be obtained by less cardiovascular active drugs. PMID- 15114344 TI - Synaptic gating and ADHD: a biological theory of comorbidity of ADHD and anxiety. AB - To derive a biologically based theory of comorbidity in Attention Deficit Hyperactivity Disorder (ADHD). Theoretical concepts and empirical studies were reviewed to determine whether the behavioral inhibition concept provided an understanding of biological processes involved in comorbidity in ADHD. Empirical studies of ADHD have shown comorbidity of ADHD and anxiety, while studies of behavioral inhibition tend to suggest independent disruptive and anxiety traits. This paradox can be resolved by an understanding of the dynamics of mesolimbic dopamine (DA) systems, where reward and delay of reinforcement are determined by tonic/phasic DA relationships, resulting in impulsive 'fearless' responses when impaired. On the other hand, comorbid anxiety is related to impaired synaptic processes, which selectively gate fear (or aggressive) responses from the amygdala at the accumbens. Monosynaptic convergence between prefrontal, hippocampal, and amygdala projection neurons at the accumbens allows the operation of a synaptic gating mechanism between prefrontal cortex (PFC), hippocampus, and amygdala. Impairment of this mechanism by lowered PFC inhibition allows greater amygdala input, and anxiety-related processes more impact, over the accumbens. In conclusion, a dual theory incorporating long-term tonic/phasic mesolimbic DA relationships and secondly impairment of PFC and hippocampal inputs to synaptic gating of anxiety at the accumbens has implications for comorbidity in ADHD, as well as for possible pharmacological interventions, utilizing either stimulant or axiolytic interventions. The use of DA partial agonists may also be of interest. PMID- 15114343 TI - Prepulse inhibition deficits in GAD65 knockout mice and the effect of antipsychotic treatment. AB - Recent postmortem studies in humans suggest that defects in GABAergic neurotransmission might contribute to the neuropathology associated with schizophrenia. Disturbances in GABAergic systems may also contribute to the sensorimotor gating deficits classically observed in schizophrenic patients, including deficits in prepulse inhibition (PPI). To explore the relationship, the current study examined the integrity of PPI and startle habituation in knockout (KO) mice that lack the GABA synthesizing enzyme glutamic acid decarboxylase 65 (GAD 65). GAD65 KO mice displayed normal baseline and habituated startle responses, which did not differ from GAD65 wild-type (WT) or heterozygous (HET) mice. However, GAD65 KO mice showed robust deficits in PPI which were reversed by the atypical antipsychotic agent clozapine. These results lend support to the view that abnormalities in GABAergic systems might contribute to the basic pathophysiological mechanisms in schizophrenia. PMID- 15114345 TI - A defeat for primate research. PMID- 15114347 TI - Eph-ephrin promiscuity is now crystal clear. PMID- 15114348 TI - Presynaptic NMDA receptors get into the act. PMID- 15114349 TI - Tlx genes make an exciting choice. PMID- 15114350 TI - Visual and action cues contribute to the self-other distinction. PMID- 15114352 TI - Limits to growth: why neuroscience needs large-scale science. PMID- 15114353 TI - Neurogenomics: at the intersection of neurobiology and genome sciences. AB - Neurogenomics is the study of how the genome as a whole contributes to the evolution, development, structure and function of the nervous system. It includes investigations of how genome products (transcriptomes and proteomes) vary in time and space. Neurogenomics differs markedly from the application of genome sciences to other systems, particularly in the spatial category, because anatomy and connectivity are paramount to our understanding of function in the nervous system. We focus here on some of the influences of genomics and its associated technologies on neuroscience. We discuss comparative genomics, gene expression atlases of the brain, network genetics and applications to behavioral phenotypes, and consider the culture, organization and funding of genome-scale projects. PMID- 15114354 TI - Progress in the use of microarray technology to study the neurobiology of disease. AB - The diverse functions of the brain are mediated by neurons and glia whose phenotype is defined by a dynamically maintained set of gene transcripts, or 'transcriptome'. Large-scale analysis of gene expression in postmortem brain using microarray technology has the potential to elucidate molecular changes that occur in disease states. There are unique challenges associated with studies of postmortem brain, including limited sample sizes and variable clinical phenotypes that are typical of complex disorders. Nevertheless, recent microarray-based studies have implicated both individual dysregulated genes and abnormal patterns of gene expression in brain disorders. PMID- 15114355 TI - Proteomics in postgenomic neuroscience: the end of the beginning. AB - Proteomics is complementary to genomic approaches anchored in DNA and RNA. Global characterization of proteins is providing new insights into general biological structures as well as synapses, receptor complexes and other neuronal and glial features. Current challenges for proteomics of the nervous system include problems relating to sample preparation, brain complexity, limited databases and informatics tools. The combination of proteomics with other global functional genomic approaches at the levels of genome and transcriptome, together with network biology, will provide important bridges between genes, physiology and pathology. PMID- 15114356 TI - Large-scale recording of neuronal ensembles. AB - How does the brain orchestrate perceptions, thoughts and actions from the spiking activity of its neurons? Early single-neuron recording research treated spike pattern variability as noise that needed to be averaged out to reveal the brain's representation of invariant input. Another view is that variability of spikes is centrally coordinated and that this brain-generated ensemble pattern in cortical structures is itself a potential source of cognition. Large-scale recordings from neuronal ensembles now offer the opportunity to test these competing theoretical frameworks. Currently, wire and micro-machined silicon electrode arrays can record from large numbers of neurons and monitor local neural circuits at work. Achieving the full potential of massively parallel neuronal recordings, however, will require further development of the neuron-electrode interface, automated and efficient spike-sorting algorithms for effective isolation and identification of single neurons, and new mathematical insights for the analysis of network properties. PMID- 15114357 TI - Using multi-neuron population recordings for neural prosthetics. AB - Classical single-neuron recording methods led to 'neuron-centric' concepts of neural coding, whereas more recent multi-neuron population recordings have inspired 'population-centric' concepts of distributed processing in neural systems. Because most neocortical neurons code information coarsely, sensory or motor processing tends to be widely distributed across neuronal populations. Dynamic fluctuations in neural population functions thus involve subtle changes in the overall pattern of neural activity. Mathematical analysis of neural population codes allows extraction of 'motor signals' from neuronal population recordings in the motor cortices, which can then be used in real-time to directly control movement of a robot arm. This technique holds promise for the development of neurally controlled prosthetic devices and provides insights into how information is distributed across several brain regions. PMID- 15114358 TI - Multiple neural spike train data analysis: state-of-the-art and future challenges. AB - Multiple electrodes are now a standard tool in neuroscience research that make it possible to study the simultaneous activity of several neurons in a given brain region or across different regions. The data from multi-electrode studies present important analysis challenges that must be resolved for optimal use of these neurophysiological measurements to answer questions about how the brain works. Here we review statistical methods for the analysis of multiple neural spike train data and discuss future challenges for methodology research. PMID- 15114359 TI - Neurobehavioral assessment in the information age. AB - The elucidation of the human and mouse genomes provides new opportunities for exploring the genetic underpinnings of complex mammalian behaviors. This information also provides new windows into the pathophysiology and treatment of neuropsychiatric diseases. Optimal use of the rapidly escalating numbers of mouse lines engineered for these purposes is hindered, however, by practical and theoretical limitations of common behavioral analyses. New strategies combining automated behavioral monitoring and information technologies are currently under development in both academic and industrial settings. These hold promise, both for improving the throughput of mouse behavioral assessment and for providing new insights into the neurobiology of mammalian behavioral regulation. PMID- 15114360 TI - E-neuroscience: challenges and triumphs in integrating distributed data from molecules to brains. AB - Imaging, from magnetic resonance imaging (MRI) to localization of specific macromolecules by microscopies, has been one of the driving forces behind neuroinformatics efforts of the past decade. Many web-accessible resources have been created, ranging from simple data collections to highly structured databases. Although many challenges remain in adapting neuroscience to the new electronic forum envisioned by neuroinformatics proponents, these efforts have succeeded in formalizing the requirements for effective data sharing and data integration across multiple sources. In this perspective, we discuss the importance of spatial systems and ontologies for proper modeling of neuroscience data and their use in a large-scale data integration effort, the Biomedical Informatics Research Network (BIRN). PMID- 15114361 TI - Sharing neuroimaging studies of human cognition. AB - After more than a decade of collecting large neuroimaging datasets, neuroscientists are now working to archive these studies in publicly accessible databases. In particular, the fMRI Data Center (fMRIDC), a high-performance computing center managed by computer and brain scientists, seeks to catalogue and openly disseminate the data from published fMRI studies to the community. This repository enables experimental validation and allows researchers to combine and examine patterns of brain activity beyond that of any single study. As with some biological databases, early scientific, technical and sociological concerns hindered initial acceptance of the fMRIDC. However, with the continued growth of this and other neuroscience archives, researchers are recognizing the potential of such resources for identifying new knowledge about cognitive and neural activity. Thus, the field of neuroimaging is following the lead of biology and chemistry, mining its accumulating body of knowledge and moving toward a 'discovery science' of brain function. PMID- 15114362 TI - Gene expression nervous system atlas (GENSAT). PMID- 15114363 TI - A genome end-game: understanding gene function in the nervous system. PMID- 15114364 TI - WebQTL: rapid exploratory analysis of gene expression and genetic networks for brain and behavior. PMID- 15114365 TI - Neurodatabase.org: networking the microelectrode. PMID- 15114366 TI - Exposure to wood dust and endotoxin in small-scale wood industries in Tanzania. AB - Workers in small-scale wood industries (SSWI) have increased risks of developing asthma and other respiratory diseases. Wood dust and microbial agents have both been suggested to play a role, but few studies have measured endotoxin exposure in SSWI in Africa. We assessed inhalable dust levels in 281 samples from 115 workers and bacterial endotoxins levels in 157 samples from 136 workers from SSWI in Dar es Salaam, Tanzania. The overall geometric mean of personal exposure was 3.3 mg/m(3); geometric standard deviation (GSD) 2.5; range 0.45-67.0 mg/m(3)) and 91 EU/m(3) (GSD 3.7; range 9-4914.8 EU/m(3)) for wood dust and endotoxins, respectively. Dust and endotoxin levels were weakly correlated (r = 0.44, n = 157, P < 0.0001). Between- and within-worker variances and percentages explained by the differences among job titles and seasons were 0.31 (9%) and 0.35 (30%), respectively, for wood dust exposure, and 0.35 (0%) and 0.35 (38%) for endotoxin exposure. Higher dust and endotoxin exposure levels were observed in the dry compared to the wet season, after correcting for differences in exposure between jobs. Carving and manual cleaning were associated with the highest dust exposures. Sewing seat covers and manual cleaning were associated with the highest endotoxin exposures. Dust and endotoxin exposure levels in SSWI are high and appropriate control measures are necessary. PMID- 15114367 TI - Genetic covariance structure of growth in the salamander Ambystoma macrodactylum. AB - The size of an organism at any point during ontogeny often has fitness consequences through either direct selection on size or through selection on size related morphological, performance, or life history traits. However, the evolutionary response to selection on size across ontogeny (a growth trajectory) may be limited by genetic correlations across ages. Here we characterize the phenotypic and genetic covariance structure of length and mass growth trajectories in a natural population of larval Ambystoma macrodactylum using function-valued quantitative genetic analyses and principal component decomposition. Most of the phenotypic and genetic variation in both growth trajectories appears to be confined to a single principal component describing a pattern of positive covariation among sizes across all ages. Higher order principal components with no significant associated genetic variation were identified for both trajectories, suggesting that evolution towards certain patterns of negative covariation between sizes across ages is constrained. The well-characterized positive relationship between size at metamorphosis and fitness in pond-breeding amphibians predicts that the across-age covariance structure will strongly limit evolution only if there is negative selection on size prior to metamorphosis. The pattern of genetic covariation observed in this study is similar to that observed in other vertebrate taxa, indicating that size may often be highly genetically and phenotypically integrated across ontogeny. Additionally, we find that phenotypic and genetic analyses of growth trajectories can yield qualitatively similar patterns of covariance structure. PMID- 15114368 TI - Quantitative trait loci affecting growth-related traits in wild barley (Hordeum spontaneum) grown under different levels of nutrient supply. AB - The genetic basis of phenotypic plasticity of relative growth rate (RGR), its components and associated morphological traits was studied in relation to nutrient limitation. In all, 140 F(3) lines from a cross, made between two Hordeum spontaneum (wild barley) accessions sampled in Israel, were subjected to growth analysis under two nutrient levels. Quantitative trait loci (QTLs) were detected for RGR and three of its components, leaf area ratio (LAR), specific leaf area and leaf mass fraction (LMF). Indications for close linkage (potential pleiotropy) were found, for example, for LAR and LMF. An interesting case is on chromosome 6, at which QTLs for RGR and seed mass were detected in the same region. These QTLs had opposite additive effects, supporting earlier results that plants growing from lighter seeds had a higher RGR. Only two QTLs were significant under both nutrient conditions, suggesting large QTL x environment interactions for most traits. For 21 out of 26 QTLs, however, the additive genetic effect was of identical sign in both nutrient environments, but reached the significance threshold in only one of them. Nevertheless, some QTLs detected in one of the two environments had virtually no effect in the other, and QTLs for plasticity were detected for RGR, LAR and LMF, as well as for some morphological traits. QTLs with opposite effects under high and low nutrients were not found. Thus, at the genetic level, there was no evidence for a trade-off between faster growth at high versus low nutrient levels. PMID- 15114369 TI - Genetic and phenotypic characterization of mutations in myosin-binding protein C (MYBPC3) in 81 families with familial hypertrophic cardiomyopathy: total or partial haploinsufficiency. AB - Mutations in the MYBPC3 gene, encoding the sarcomere protein myosin-binding protein C, are among the most frequent causes of autosomal dominant familial hypertrophic cardiomyopathy (FHC). We studied the frequency, type, and pathogenetic mechanism of MYBPC3 mutations in an unselected cohort of 81 FHC families, consecutively enrolled at a tertiary referral center. Nine mutations, six of which were novel, were found in 10 (12.3%) of the families using single strand conformation polymorphism and DNA sequencing. A frameshift mutation in exon 2 clearly suggests that haploinsufficiency is a pathogenetic mechanism in FHC. In addition, splice site mutations in exon 6 and intron 31, a deletion in exon 13, and a nonsense mutation in exon 25, all lead to premature termination codons, most likely causing loss of function and haploinsufficiency. Furthermore, there were two missense mutations (D228N and A833 T) and one in-frame deletion (DeltaLys813). A considerable intrafamilial variation in phenotypic expression of MYBPC3-based FHC was noted, and we suggest that mutations influencing stability of mRNA could play a role in the variable penetrance and expressivity of the disease, perhaps via partial haploinsuffciency. PMID- 15114370 TI - Association between a polymorphism in the carboxyl ester lipase gene and serum cholesterol profile. AB - Carboxyl ester lipase (CEL) is involved in the hydrolysis and absorption of dietary lipids, but it is largely unknown to what extent CEL could be involved in determining the serum lipid levels. The C-terminal part of CEL consists of a unique structure with proline-rich O-glycosylated repeats of 11 amino-acid residues each. The common variant of the human CEL gene contains 16 proline-rich repeats, but there is a high degree of polymorphism in the repeated region. While the biological function of the polymorphic repeat region is unknown, it has been suggested that it may be important for protein stability and/or secretion of the enzyme. Given that the polymorphism in the repeated region may affect the functionality of the protein, this study aimed to investigate whether the number of repeated units is correlated to serum lipid phenotype. Comparison of CEL repeat genotype and serum lipid phenotype revealed an association between the number of repeats and serum cholesterol profile. Individuals carrying at least one allele with fewer than the common 16 repeats had significantly lower total and low-density lipoprotein (LDL) cholesterol levels compared to individuals carrying two common alleles. This gives support to the notion that CEL may be involved in determining the plasma lipid composition. PMID- 15114371 TI - Genetic isolates in Corsica (France): linkage disequilibrium extension analysis on the Xq13 region. AB - Genetic isolates with a history of a small founder population, long-lasting isolation and population bottlenecks represent exceptional resources in the identification of genes involved in the pathogenesis of multifactorial diseases. In these populations, the disease allele reveals linkage disequilibrium (LD) with markers over significant genetic intervals, therefore facilitating disease locus identification. This study has been designed to examine the background LD extension in some subpopulations of Corsica. Our interest in the island of Corsica is due to its geographical and genetic proximity to the other Mediterranean island of Sardinia. Sardinian isolates in which the extension of the background LD is particularly high have been recently identified and are now the object of studies aimed at the mapping of genes involved in complex diseases. Recent evidence has highlighted that the genetic proximity between the populations of Corsica and Sardinia is particularly true for the internal conservative populations. Given these considerations, Sardinia and Corsica may represent a unique system to carry out parallel association studies whose results could be validated by comparison. In the present study, we have analyzed the LD extension on the Xq13 genomic region in three subpopulations of Corsica: Corte, Niolo and Bozio, all located in the mountainous north-center of the island. Our results show a strong degree of LD over long distance for the population of Bozio and to a less extent for the population of Niolo. Their LD extent is comparable to or higher than that reported for other isolates. PMID- 15114372 TI - Systemic lupus erythematosus (SLE) and chromosome 16: confirmation of linkage to 16q12-13 and evidence for genetic heterogeneity. AB - Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease with significant morbidity and mortality, characterized by remarkable clinical variability with unknown etiology. Genetic contribution to the development of SLE is well established. Recently, we found evidence (P<0.004) of linkage at 16p13 and 16q12-13 in a genome scan based on 37 Hispanic families. The main objective of this study is to replicate and confirm the linkage at these two genomic locations in two large independent replication data sets designated as, group-1 and group-2, consisting of 172 and 120 multiplex SLE families, respectively. We have found a significant evidence of linkage with high heterogeneity (HLOD=4.85, alpha=35%) at 16q12-13 in group-2. Other independent research groups also reported the SLE susceptibility at or close to 16q12-13 previously. Therefore, independent published reports, together with our initial linkage with Hispanics and followed by significant evidence from group-2, provide a strong and confirmed evidence for an SLE susceptibility locus at 16q12-13. PMID- 15114373 TI - BRCA1 and sex ratio. AB - Two recent papers suggest distorted sex and transmission ratios associated with BRCA1 mutations. If real, these would provide novel insights into the normal biological function of this gene and have implications for genetic epidemiologic methods used to estimate penetrance. We addressed these observations in two settings: offspring of 283 mutation carriers and 471 mutation negative subjects from BRCA1/2 mutation-positive families with multiple cases of breast and ovarian cancer (NCI families); and relatives of 115 BRCA1/2 mutation carriers from the Washington Ashkenazi Study (WAS). The male:female ratio was below one in both BRCA1 (0.85, 95% CI 0.7-1.1 in NCI families; 0.90, 95% CI 0.6-1.4 in WAS) and BRCA2 families (0.77, 95% CI 0.5-1.3 and 0.80, 95% CI 0.5-1.2, in the NCI and WAS study groups, respectively). None of the sex ratios deviated significantly from one, and there was no significant difference between BRCA1 and BRCA2 families. The reduced sex ratio was due largely to the offspring of males, a distortion that is probably an artifact of ascertainment biases. Among adult daughters without breast or ovarian cancer born to mutation carriers, as expected, fewer than 50% were mutation carriers (39% in BRCA1 families and 44% in BRCA2 families). It is difficult, due to ascertainment biases, to draw firm conclusions regarding sex ratios in studies of a sex-limited phenotype. Nonetheless, these observations do not support the idea that BRCA1 mutation carriers have a lower ratio of male offspring than BRCA2 mutation carriers. PMID- 15114374 TI - Application of microarrays to the analysis of the inactivation status of human X linked genes expressed in lymphocytes. AB - Dosage compensation in mammalian females is achieved by the random inactivation of one X chromosome early in development; however, inactivation is not complete. In addition to a majority of pseudoautosomal loci, there are genes that are expressed from both the active and the inactive X chromosomes, and which are interspersed among other genes subject to regular dosage compensation. The patterns of X-linked gene expression in different tissues are of great significance for interpreting their impact on sex differences in development. We have examined the suitability and sensitivity of a microarray approach for determining the inactivation status of X-linked genes. Biotinylated cRNA from six female and six male lymphocyte samples were hybridised to Affymetrix HG-U133A microarrays. A total of 36 X-linked targets detected significantly higher levels of female transcripts, suggesting that these corresponded to sequences from loci that escaped, at least partly, from inactivation. These included genes for which previous experimental evidence, or circumstantial evidence, existed for their escape, and some novel candidates. Six of the targets were represented by more than one probe set, which gave independent support for the conclusions reached. PMID- 15114375 TI - TEAM: a tool for the integration of expression, and linkage and association maps. AB - The identification of genes primarily responsible for complex genetic disorders is a daunting task. Despite the assignment of many susceptibility loci, there has only been limited success in identifying disease genes based solely on positional information from genome-wide screens. The incorporation of several complementary strategies in a single integrated approach should facilitate and further enhance the efficacy of this search for genes. To permit the integration of linkage, association and expression data, together with functional annotations, we have developed a Java-based software tool: TEAM (tool for the integration of expression, and linkage and association maps). TEAM includes a genome viewer, capable of overlaying karyobands, genes, markers, linkage graphs, association data, gene expression levels and functional annotations in one composite view. Data management, analysis and filtering functionality was implemented and extended with links to the Ensembl, Unigene and Gene Ontology databases to facilitate gene annotation. Filtering functionality can help prevent the exclusion of poorly annotated, but differentially expressed, genes that reside in candidate regions that show linkage or association. Here we demonstrate the program's functionality in our study on coeliac disease (OMIM 212750), a multifactorial gluten-sensitive enteropathy. We performed a combined data analysis of a genome-wide linkage screen in 82 Dutch families with affected siblings and the microarray expression profiles of 18,110 cDNAs in 22 intestinal biopsies. PMID- 15114376 TI - Double-blind, randomized feedback control fails to improve the hypocholesterolemic effect of a plant-based low-fat diet in patients with moderately elevated total cholesterol levels. AB - OBJECTIVE: To determine whether the cholesterol-lowering effect of a plant-based low-fat diet can be improved by a flexible control design that controls the extent of fat reduction based on the individual response of blood cholesterol. DESIGN: Randomized, double-blind intervention study. SETTING: A hotel in Prerow, Germany. SUBJECTS: A total of 32 participants (21 female and 11 male participants) with total cholesterol level > 5.7 mmol/l. INTERVENTION: The control group consumed a plant-based low-fat diet with constantly 20% of energy as fat; the intervention group received a diet with either 20 or 15% of energy as fat, depending on the serum cholesterol response of the preceding week. A flexible control design based on the individual cholesterol response during a run in period of 1 week was used within a low-fat intervention. RESULTS: During the run-in period, the consumption of a plant-based low-fat diet led to a reduction in total cholesterol by 18+/-6 mmol/l (P < 0.001), in LDL cholesterol by 19+/-9 mmol/l (P < 0.001) and triglycerides by 13+/-3 mmol/l (P < 0.001). During the feedback control period, an additional reduction in total cholesterol by 13+/-8 (P < 0.001) and in LDL cholesterol by 17+/-11 (P < 0.001) was observed compared to 15+/-15 and 7+/-18 in the control group. The effect of an additional feedback control was only marginal and not statistically significant compared to the effect of the low-fat diet alone. CONCLUSIONS: On a level of fat intake already reduced to 20% of energy, the use of a feedback control to adapt the fat content of the diet depending on the individual serum cholesterol response was not more effective in reducing blood cholesterol levels than a plant-based low-fat diet alone. PMID- 15114377 TI - Calcium and vitamin D3 supplements in calcium and vitamin D3 sufficient early postmenopausal healthy women. AB - OBJECTIVE: To study the calcium homeostasis in healthy, calcium and vitamin D replete early postmenopausal women during oral supplementation with calcium and vitamin D3. DESIGN: A prospective, placebo-controlled, randomised, double-single blind, 3-week study. SETTING: Outpatient clinic at Copenhagen University Hospital, Denmark. SUBJECTS: In all, 17 started, one was excluded. Totally, 16 healthy women, 45-61 y of age (mean 57.3 y) who were at least 4 y after menopause (mean 6.7 y) completed. INTERVENTIONS: All underwent three consecutive 7-day study periods. Each began with 4 days of normal diet followed by 3 days treatment of either C: one tablet of 1.250 mg calcium carbonate (ie 500 mg Ca2+ per tablet) twice daily (breakfast and dinner), or CD3: as in C but plus 400 IU vitamin D3 b.i.d., or P (only) placebo tablets b.i.d. RESULTS: At baseline plasma 25 hydroxycholecalciferol was normal (66+/-22 nmol/l) and the calcium intake without supplements 850 mg/day. In group C, the 24-h urinary calcium increased by 35% (6.9+/-2.0 mmol), vs the placebo group P (5.1+/-1.6 mmol) (P < 0.05). Addition of 800 IU vitamin D3 daily (CD3) did not increase calcium excretion further (6.6+/ 2.1 mmol) but decreased plasma 1,25-(OH)2-vitamin D3 by 21% (P < 0.05). CONCLUSIONS: In this carefully controlled study calcium plus vitamin D3 supplements only had minor influences of uncertain significance on the calcium balance in healthy, calcium and vitamin D sufficient early postmenopausal women. PMID- 15114378 TI - [Unnecessary overlapping and old news]. PMID- 15114379 TI - [Should PET scanning be used in lung cancer?]. PMID- 15114380 TI - [New technology--difficult evaluation]. PMID- 15114381 TI - [Knee luxation--follow-up after surgery]. AB - BACKGROUND: Traumatic dislocation is the most severe ligamentous injury of the knee. The traditional definition of knee dislocations has been expanded to include bicruciate knee injuries with one or both lateral ligaments injured, even when the knee is reduced on initial presentation. Surgical treatment is internationally well accepted. MATERIAL AND METHODS: From 1996 to 2002, 87 patients with knee dislocation were treated in our department. In this prospective study, 55 patients were followed up with functional knee tests a minimum of one year (mean: 3.5 years) after surgical treatment. RESULTS: 59% of the injuries were high-energy traumas. Mean age was 34 (12-80). 57% of the patients achieved excellent to good results on the Lysholm Score. INTERPRETATION: Positive prognostic factors are likely to be age less than 40 at the time of the accident or sports-related injury. Fifty-four of the 55 patients in the follow-up study had returned to work or studies after one year. Dislocated knee is not only related to high-energy and traffic injuries; it also occurs in low-energy trauma and sports injuries. PMID- 15114382 TI - [Gallbladder surgery in a district hospital--a prospective registration]. AB - BACKGROUND: Health authorities and patients are expected to be more active in inquiring about the outcome of surgery for each specific hospital. Prospective registration performed in a district hospital has to go on for years until the results can be evaluated. Specific data for gallbladder surgery performed in Norwegian district hospitals have not been reported before. MATERIAL AND METHODS: All 220 cholecystectomies carried out from May 1993 to March 2001 were registered prospectively. RESULTS: Two surgeons performed 93% of the operations with an average annual operator volume of 28. The operation rate was 8.5 per 10,000 inhabitants. The need for conversion was 9%. The peri and postoperative complication rate for laparoscopic and converted operations was 11%. INTERPRETATION: Our operation rate is somewhat higher than the average in Norway. The number of annual operations in a district hospital is somewhat low, but as almost all procedures are carried out by a limited number of surgeons, operator volume is compatible with data from larger hospitals in Norway. The need for conversion and the rate and seriousness of complications peri and postoperatively does not seem to be higher in this district hospital than in county and university hospitals in Norway. PMID- 15114383 TI - [Apheresis in severe hypercholesterolaemia]. AB - BACKGROUND: Hypercholesterolaemia is usually successfully treated with statins but in some cases the medication has insufficient cholesterol-lowering effect or is not well tolerated. In these instances LDL apheresis is an option. MATERIAL AND METHODS: We share some of our own experience with this treatment and then review the literature regarding LDL apheresis. RESULTS: LDL apheresis seems a safe and effective way of lowering LDL cholesterol. Mortality and morbidity is reduced in selected patient groups. Quite probably, too few Norwegian patients are offered this treatment. INTERPRETATION: LDL apheresis should be available in all health regions of Norway and the treatment should be known to all those who treat severe hypercholesterolaemia or coronary artery disease. PMID- 15114384 TI - [Positron emission tomography in the diagnosis of lung cancer]. AB - BACKGROUND: Lung cancer is a common malignancy. Positron emission tomography (PET) has become an important tool in the staging of the disease with regard to operability and in order to determine radiation volume when surgery is not indicated. MATERIALS AND METHODS: We have assessed and discuss results recently published in major international journals. RESULTS: In approximately 60% of the patients, the planning of treatment is modified after a PET investigation. INTERPRETATION: In contrast to the state the art in most industrialised countries, dedicated PET is not available in Norway. Hence lung cancer is suboptimally treated. PMID- 15114385 TI - [Knowledge about mental retardation among psychiatric residents]. AB - BACKGROUND: Since the deinstitutionalisation reform in 1991, the general health services, including psychiatry, are expected to serve the needs of people with mental retardation. Studies have shown that users generally find the service provided by psychiatry unsatisfactory. One reason for this may be a lack of confidence on the part of psychiatrists to take on such work. We focus on whether future psychiatrists feel that they learn enough about this topic. MATERIAL AND METHODS: We asked 60 residents in psychiatry in various stages of their graduate medical education to evaluate their own knowledge or skills in 109 fields expected to be mastered by a specialist in psychiatry. Items with high relevance to mental retardation were compared with other items of similar relevance to psychiatric practice. RESULTS: The residents felt they lacked skills/knowledge in psychiatric and medicolegal fields mainly concerning people with mental retardation. In fields that were highly relevant for both general psychiatric practice and mental retardation the residents reported higher confidence. INTERPRETATION: The efforts during the reform process to secure psychiatric services for people with mental retardation are not reflected in the training of new psychiatrists or in their confidence in their ability to undertake such work. PMID- 15114386 TI - [Gitelman's syndrome--a differential diagnosis in hypokalemia]. AB - BACKGROUND: Gitelman's syndrome is a rare disease characterised by low levels of potassium and magnesium in the blood. It is caused by mutations in the gene encoding the thiazide-sensitive sodium chloride cotransporter in the distal collecting duct. MATERIAL AND METHODS: We present four patients (two brothers and two sisters) with Gitelman's syndrome and review the literature regarding the disease. RESULTS AND INTERPRETATION: Gitelman's syndrome should be considered in patients with persistently low levels of potassium and magnesium. The diagnosis is confirmed by genetic testing. PMID- 15114387 TI - [Vascular dementia--an ill-defined concept]. AB - BACKGROUND: Vascular dementia is defined as dementia caused by cerebrovascular disease and is seen as the most common cause of dementia after Alzheimer's disease. In our opinion, vascular dementia is an ill-defined term; this proposition is discussed in this article. MATERIAL AND METHODS: The concept of vascular dementia is discussed based on the literature and our own clinical experience. RESULTS: The prevalence of vascular dementia varies markedly across different studies and several clinical diagnostic criteria are used. There has been an ongoing debate as to which radiological changes are associated with vascular dementia; we question the use of neuropathological changes as a diagnostic gold standard. Several studies have shown similarities between dementia of the Alzheimer type and vascular dementia regarding risk factors and symptomatology. INTERPRETATION: We conclude that the concept of vascular dementia should be re-evaluated. It is important to establish valid and reliable criteria for vascular dementia, especially in the light of the therapeutic consequences. PMID- 15114388 TI - [Disagreement about epilepsy and driving]. AB - BACKGROUND: To find out how current regulations concerning epilepsy and driving licence are applied in clinical practice in Norway. MATERIAL AND METHODS: A questionnaire was sent to all 344 members of the Norwegian Neurological Association. RESULTS: 56% responded within 3 months, most of them (152) working in the field of clinical neurology. The majority (73%) did not see reporting a "situation-related" seizure to the authorities as obligatory, but almost all of these respondents (94%) would temporarily disallow driving. 60% did not consider it obligatory to report the exclusive occurrence of simple partial seizures with retained consciousness and motor control. A surprisingly large number (36%) would never recommend dispensation from the rule instituting a one-year driving ban. 39% generally used EEG in the assessment of driving fitness, whereas 17 % rarely did so. Only 50% instructed their patients not to drive during antiepileptic drug withdrawal. 45% did not consider seizure recurrence resulting from consented withdrawal to activate the obligation to report. CONCLUSION: There is poor consensus among neurologists on many aspects of driving and epilepsy, particularly on the assessment of risk when drugs are withdrawn. These issues should be focused on in graduate and postgraduate education in clinical neurology. PMID- 15114389 TI - [Acute coronary syndrome in patients using acetylsalicylic acid or warfarin]. PMID- 15114390 TI - [Modern diagnosis and treatment of heart failure]. AB - Chronic heart failure is a prevalent condition associated with high morbidity, high mortality and reduced quality of life. Chronic heart failure is the end stage of various forms of heart disease. The prognosis is poor, worse than for patients with various forms of cancer. The diagnosis of heart failure is made in the presence of multiple symptoms and signs combined with objective evidence of cardiac dysfunction. Treatment consists of pharmacological as well as non pharmacological approaches. ACE inhibitors and beta-blockers are the basis of treatment, while diuretics and other medications are given on an individual basis. The aim of treatment is to reduce progression of the underlying disease and to reduce mortality and morbidity. PMID- 15114391 TI - [Cardiac resynchronisation therapy by biventricular pacing in severe heart failure]. AB - Biventricular pacing with one pacemaker lead introduced via the coronary sinus to a coronary vein and one pacemaker lead located endocardially at the apex of the right ventricle is used for cardiac resynchronisation therapy in patients with severe heart failure and bundle branch block indicating asynchronous contraction of the left ventricle. Randomised multicentre studies show significant improvement of functional capacity, quality of life and left ventricular systolic function in patients with severe heart failure and bundle branch block. In one study, a significant reduction of 77% of hospitalisation days has been demonstrated. Combined implantable biventricular pacing and cardioverter defibrillator units resulted in a 43% reduction of mortality in patients with severe heart failure compared to optimal pharmacological treatment only. Our studies show that selection of patients for cardiac resynchronisation therapy by using echocardiographic methods in addition to ECG may help to avoid implanting pacemakers in patients who will not respond to the therapy. In our studies resynchronisation of the contraction of the left ventricle was demonstrated by using colour tissue Doppler measurements. The mechanisms for the haemodynamic benefit of cardiac resynchronization therapy are discussed. This relatively new therapy represents a significant progress in the treatment of patients with severe heart failure and asynchronous left ventricular contraction refractory to treatment. PMID- 15114392 TI - [Heart transplantation]. AB - The first heart transplantation in the Nordic countries was performed at Rikshospitalet in Oslo, Norway in 1983 and the method is now accepted as a good treatment for selected patients. In this article we present a review of heart transplantation as it is practiced in our hospital. Because of donor shortage, the selection criteria are strict. It is most important that the patients are motivated and able to cooperate. They should be in heart failure class IV (III IV) on the best medical treatment with no other treatment alternatives. Estimated survival without transplantation should be less than 6-12 months. Important contraindications are concurrent diseases with a more severe prognosis than that expected after transplantation. One-year and ten-year survival after transplantation at Rikshospitalet are 85% and 53%. The most important causes of death in the early postoperative period are rejection and infections. Later, the most frequent causes of death are accelerated coronary artery disease and cancer. Collaboration between the referring physician and the transplant centre is essential for a better prognosis for heart failure and after transplantation. PMID- 15114393 TI - [Complications in ear-nose-throat day surgery]. AB - BACKGROUND: More and larger operations are performed on an outpatient basis with patient leaving the hospital the same day and complications may remain unknown to the hospital unless the patients are asked explicitly. MATERIAL AND METHODS: Complications from 383 ear-nose-throat operations were registered continuously by the surgeon and supplemented with data from postoperative consultations and reports from other hospitals or doctors, and then compared with data from 328 (86%) postoperative patient interviews. RESULTS: According to the patient interviews, the rate of complications (16.7%) was much higher than the hospital was aware of (4.7%). Most unknown complications (40 out of 64) were mild but five were serious enough to require reoperation under general anesthesia. INTERPRETATION: Registration of surgical complications should include postoperative interviews with patients. PMID- 15114394 TI - [Approaching the locked dialogues of the body]. AB - BACKGROUND: The objective of this project was to develop a clinical communication tool to change locked dialogues between doctor and patient in general practice consultations. The target group was patients suffering from long-standing symptoms without clinical findings. Patients were encouraged to keep an illness diary or to write personal notes on their illness as a tool for cooperating with the doctor. MATERIAL AND METHODS: The material was collected from 36 consultations with 16 patients. The qualitative evaluation aimed at giving a description and analysis of how the illness diary affected the doctor-patient dialogue in the consultation. The analysis was modified from phenomenology-based methodology. A part of the project dealt with the methodological problems of doing research on an interaction in which the general practitioner is both participant (doctor) and observer (researcher). RESULTS AND INTERPRETATION: The diary enhanced the doctor's basis for clinical decision making by giving medically relevant information from the patient and by producing more concrete information. The diary gave the doctor access to patients' insight in their own problems. The clinical dialogue modified the patient's negative internal monologues in order to search for alternate dialogues as a basis for personal coping strategies. The project demonstrated that written notes might give patient and doctor a new understanding of a previously spoken message. The writing procedure gave patients an opportunity to reflect upon their illness presentation. PMID- 15114396 TI - [Private engagement concerning the HIV]. PMID- 15114398 TI - [Cardiology as an engine for internal medicine]. PMID- 15114399 TI - [Norwegian cardiology and Norwegian general practice]. PMID- 15114400 TI - [Wanted]. PMID- 15114401 TI - [Children as research subjects]. PMID- 15114404 TI - [Children and confidentiality]. PMID- 15114405 TI - [Information and consent in the treatment of children and adolescents]. PMID- 15114406 TI - [Response time of ambulances]. PMID- 15114408 TI - [Multi-dose packaging is a good system]. PMID- 15114409 TI - [How should the resource prioritization be performed?]. PMID- 15114410 TI - Effect of cyclosporine A on the tissue distribution and pharmacokinetics of etoposide. AB - PURPOSE: Cyclosporine A (CyA) is able to inhibit P-glycoprotein (P-gp) and to increase cytotoxicity of some anticancer drugs, including etoposide. However, the effect of CyA on the distribution of etoposide in normal tissues, which could affect their toxicity, has not been studied extensively. The purpose of this study was to investigate the effect of CyA on the pharmacokinetics and tissue distribution of etoposide in rats. METHODS: Etoposide was administered as an i.v. bolus injection (3 mg) or as a constant-rate i.v. infusion (8 mg/h) 1 h after the beginning of infusion of CyA or saline. Animals were killed 1 h after the bolus administration or after the beginning of infusion of etoposide, and plasma and tissue (testicle, muscle, heart, lung, spleen, kidney, liver, colon, and intestine) concentrations of etoposide, blood concentrations of CyA were determined. All analyses were performed by HPLC. RESULTS: Infusion of CyA (1 mg/h) in rats treated with an i.v. bolus of etoposide caused a decrease in the plasma clearance (5.4+/-2.1 vs 9.3+/-2.4 ml/min), and an increase in plasma and tissue concentrations of etoposide, but the tissue-to-plasma concentration ratios of etoposide were not affected. When etoposide was infused at a constant rate to reach a steady-state plasma level, coinfusion of CyA (10 mg/h) also caused a decrease in the plasma clearance (4.8+/-1.5 vs 9.8+/-4.7 ml/min), and an increase in plasma and tissue concentrations of etoposide. Only lung and spleen showed tissue-to-plasma ratios of etoposide significantly higher than obtained in rats coinfused with saline, but the differences were small. CONCLUSIONS: The higher tissue concentrations of etoposide caused by CyA administration were mainly a direct consequence of the higher plasma concentration resulting from a decrease in the clearance of etoposide rather than a consequence of changes in the tissue distribution of etoposide. Extrapolation of the results obtained in rats to clinical practice suggests that the coadministration of etoposide and CyA would not lead to an increase in the toxicity of etoposide if the dose were decreased in the same proportion as clearance of etoposide is decreased by CyA administration. PMID- 15114411 TI - Phase I/II trial of topotecan given as continuous infusion in combination with oxaliplatin in 5-FU-pretreated patients with colorectal cancer. AB - BACKGROUND: Oxaliplatin and topotecan are novel options for a variety of neoplasms. Topotecan has shown fewer side effects and higher efficacy when given as a continuous i.v. infusion compared to single doses, but this regimen has not yet been combined with oxaliplatin. PATIENTS AND METHODS: This phase I/II trial was designed to establish the dose-limiting toxicity of a combination of oxaliplatin (85-130 mg/m2 on day 1) and a continuous infusion of topotecan (initial 0.9 mg/m2 over 72-120 h). Eligible patients with metastatic colorectal cancer had progressive disease during, or within 12 weeks after, palliative fluoropyrimidine-based chemotherapy or in whom intolerable 5-FU toxicity had developed. RESULTS: The study included 21 patients. Subjectively the treatment was well tolerated but haematological toxicity was observed with the initial treatment schedule of oxaliplatin 85 mg/m2 on day 1 and topotecan 0.9 mg/m2 on days 1-5. Reducing topotecan to 0.9 mg/m2 on days 1-3 resulted also in acceptable haematological toxicity. In patients completing three or more therapy cycles, median progression-free survival was 5 months, and 50% had stable disease or showed a partial response. CONCLUSION: The recommended dose of this combination for further testing is oxaliplatin 85 mg/m2 on day 1 and topotecan 0.9 mg/m2 per day as a continuous infusion on days 1-3. PMID- 15114412 TI - Clinical images of osteopathia striata. PMID- 15114413 TI - Evaluation of anterior pituitary gland volume in childhood using three dimensional MRI. AB - BACKGROUND: Three-dimensional MRI (3D-MRI) is a reliable tool for the evaluation of anatomical volumes. Volumetric measurement of the normal anterior pituitary gland in childhood has been performed in the past by 2D-MRI calculations, but has inherent inaccuracies. OBJECTIVE: To obtain accurate normal anterior pituitary gland volume in childhood using 3D-MRI coronal sections. MATERIALS AND METHODS: The anterior pituitary gland was measured using coronal T1-weighted 3D-gradient echo sequences (section thickness 0.75 mm). The study group was composed of 95 prepubertal children (age range 2 months-10 years) with clinically normal pituitary function and no pituitary or brain abnormalities. RESULTS: A measurement error of 0.2-0.4% was assessed by using a phantom study. Volumetric evaluation of the anterior pituitary gland showed progressive growth of the gland from a mean 131+/-24 mm(3) at 2-12 months, to 249+/-25 mm(3) at 1-4 years and 271+/-29 mm(3) at 5-10 years. CONCLUSIONS: These data may be useful for paediatricians in the evaluation of patients with neuroendocrine diseases, in particular growth hormone deficiency. PMID- 15114414 TI - Congenital abdominal aortic aneurysm and renal dysplasia. AB - We report a 6-month-old boy with abdominal aortic aneurysm and dysplastic kidneys. To the best of our knowledge, this is the first report of abdominal aortic aneurysm associated with dysplastic kidneys. Serial US, MR angiography and scintigraphy were performed. No underlying cause of the aneurysm was identified. Spontaneous thrombosis occurred within 2.5 years of diagnosis, but there was persistent renovascular hypertension and progressive deterioration of renal function. PMID- 15114416 TI - Lateral transfer and recompartmentalization of Calvin cycle enzymes of plants and algae. AB - Certain Calvin cycle enzymes also function in glycolysis or gluconeogenisis, thus photosynthetic eukaryotes would be predicted to have ancestrally possessed cytosolic homologues of these enzymes derived from the eukaryotic host and plastid homologues from the cyanobacterial endosymbiont. In practice, the evolutionary histories of these enzymes are often more complex. Focusing on eukaryotes with secondary plastids, we have examined the evolution of four such genes: class I and II fructose bisphosphate aldolase (FBA), sedoheptulose bisphosphatase (SBPase), and fructose bisphosphatase (FBPase). We show that previously observed distributions of plastid and cytosolic homologues are not always found in algae with secondary plastids: there is evidence for multiple events of both lateral gene transfer and retargeting to a new cellular compartment for both cytosolic and plastid enzymes of plants and algae. In particular, we show that a clade of class II FBAs spans a greater diversity of eukaryotes that previously recognized and contains both plastid-targeted (Phaeodactylum, Odontella) and cytosolic (ascomycetes, oomycetes, Euglena, and Bigelowiella) forms. Lateral transfer events also gave rise to a subset of plant cytosolic FBA, as well as cytosolic FBPase in Toxoplasma and other coccidian apicomplexa. In contrast, it has recently been suggested that the Trypanosoma FBA and SBPase are derived from a plastid, however, greater taxonomic sampling shows that these enzymes provide no evidence for a plastid-containing ancestor of Trypanosoma. Altogether, the evolutionary histories of the FBA and SBPase/FBPase gene families are complex, including extensive paralogy, lateral transfer, and retargeting between cellular compartments. PMID- 15114417 TI - Complete mtDNA of Ciona intestinalis reveals extensive gene rearrangement and the presence of an atp8 and an extra trnM gene in ascidians. AB - The complete mitochondrial genome (mtDNA) of the model organism Ciona intestinalis (Urochordata, Ascidiacea) has been amplified by long-PCR using specific primers designed on putative mitochondrial transcripts identified from publicly available mitochondrial-like expressed sequence tags. The C. intestinalis mtDNA encodes 39 genes: 2 rRNAs, 13 subunits of the respiratory complexes, including ATPase subunit 8 ( atp8), and 24 tRNAs, including 2 tRNA-Met with anticodons 5'-UAU-3'and 5'-CAU-3', respectively. All genes are transcribed from the same strand. This gene content seems to be a common feature of ascidian mtDNAs, as we have verified the presence of a previously undetected atp8 and of two trnM genes in the two other sequenced ascidian mtDNAs. Extensive gene rearrangement has been found in C. intestinalis with respect not only to the common Vertebrata/Cephalochordata/Hemichordata gene organization but also to other ascidian mtDNAs, including the cogeneric Ciona savignyi. Other features such as the absence of long noncoding regions, the shortness of rRNA genes, the low GC content (21.4%), and the absence of asymmetric base distribution between the two strands suggest that this genome is more similar to those of some protostomes than to deuterostomes. PMID- 15114418 TI - The evolutionary history of nitrogen fixation, as assessed by NifD. AB - The evolutionary history of nitrogen fixation has been vigorously debated for almost two decades. Previous phylogenetic analyses of nitrogen fixation genes (nif) have shown support for either evolution by vertical descent or lateral transfer, depending on the specific nif gene examined and the method of analyses used. The debate centers on the placement and monophyly of the cyanobacteria, proteobacteria, and Gram-positive bacteria (actinobacteria and firmicutes). Some analyses place the cyanobacteria and actinobacteria within the proteobacteria, which suggests that the nif genes have been laterally transferred since this topology is incongruent with ribosomal phylogenies, the standard marker for comparison. Other nif analyses resolve and support the monophyly of the cyanobacteria, proteobacteria, and actinobacteria, supporting vertical descent. We have revisited these conflicting scenarios by analyzing nifD from an increased number of cyanobacteria, proteobacteria, and Gram-positive bacteria. Parsimony analyses of amino acid sequences and maximum likelihood analysis of nucleic acid sequences support the monophyly of the cyanobacteria and actinobacteria but not the proteobacteria, lending support for vertical descent. However, distance analysis of nucleic acid sequences placed the actinobacteria within the proteobacteria, supporting lateral transfer. We discuss evidence for both vertical descent and lateral transfer of nitrogen fixation. PMID- 15114419 TI - Salmonid opsin sequences undergo positive selection and indicate an alternate evolutionary relationship in oncorhynchus. AB - Positive selection can be demonstrated by statistical analysis when non synonymous nucleotide substitutions occur more frequently than synonymous substitutions (dN>dS). This pattern of sequence evolution has been observed in the rhodopsin gene of cichlids. Mutations in opsin genes resulting in amino acid (AA) replacement appear to be associated with the evolution of specific color patterns and the evolution of courtship behaviors. Within fish, AA replacements in opsin proteins have improved vision at great depths and have occurred in deep sea species. Salmonids experience diverse photic environments during their life history. Furthermore, sexual selection has resulted in species-specific male and female coloration during spawning. To look for evidence of positive selection in salmonid opsins, we sequenced the RH1, RH2, LWS, SWS1, and SWS2 genes from six Pacific salmon species as well as the Atlantic salmon. These salmonids include landlocked and migratory species and species that vary in their coloration during spawning. In each opsin gene comparison from all species sampled, traditional dN:dS analysis did not indicate positive selection. However, the more sensitive Creevey-McInerney statistical analysis indicates that RH1 and RH2 experienced positive selection early in the evolution and speciation of salmonids. PMID- 15114420 TI - Extensive gene order rearrangement in the mitochondrial genome of the centipede Scutigera coleoptrata. AB - We describe the complete mitochondrial genome of the house centipede Scutigera coleoptrata. Its gene order is unique among characterized arthropod mitochondrial genomes. Comparison to the gene order in the horseshoe crab mtDNA implies 10 or more translocations. By extending comparisons to 30 arthropod mitochondrial genomes plus two outgroups, we identify two different patterns of gene order change. The first, only affecting position and orientation of tRNAs, is much more frequent than the second, which also involves protein encoding and ribosomal genes. The analysis of the same data set using available algorithms for phylogenetic reconstruction based on gene order results in unreliable trees. This indicates that the current methods for analyzing gene order rearrangement are not suitable for wide-ranging phylogenetic studies. PMID- 15114421 TI - Dating the monocot-dicot divergence and the origin of core eudicots using whole chloroplast genomes. AB - We estimated the dates of the monocot-dicot split and the origin of core eudicots using a large chloroplast (cp) genomic dataset. Sixty-one protein-coding genes common to the 12 completely sequenced cp genomes of land plants were concatenated and analyzed. Three reliable split events were used as calibration points and for cross references. Both the method based on the assumption of a constant rate and the Li-Tanimura unequal-rate method were used to estimate divergence times. The phylogenetic analyses indicated that nonsynonymous substitution rates of cp genomes are unequal among tracheophyte lineages. For this reason, the constant rate method gave overestimates of the monocot-dicot divergence and the age of core eudicots, especially when fast-evolving monocots were included in the analysis. In contrast, the Li-Tanimura method gave estimates consistent with the known evolutionary sequence of seed plant lineages and with known fossil records. Combining estimates calibrated by two known fossil nodes and the Li-Tanimura method, we propose that monocots branched off from dicots 140-150 Myr ago (late Jurassic-early Cretaceous), at least 50 Myr younger than previous estimates based on the molecular clock hypothesis, and that the core eudicots diverged 100-115 Myr ago (Albian-Aptian of the Cretaceous). These estimates indicate that both the monocot-dicot divergence and the core eudicot's age are older than their respective fossil records. PMID- 15114422 TI - Origin of Toll-like receptor-mediated innate immunity. AB - Toll-related receptors (TLR) have been found in four animal phyla: Nematoda, Arthropoda, Echinodermata, and Chordata. No TLR has been identified thus far in acoelomates. TLR genes play a pivotal role in the innate immunity in both fruit fly and mammals. The prevailing view is that TLR-mediated immunity is ancient. The two pseudocoelomate TLRs, one each from Caenorhabditis elegans and Strongyloides stercoralis, were distinct from the coelomate ones. Further, the only TLR gene (Tol-1) in Ca. elegans did not appear to play a role in innate immunity. We argue that TLR-mediated innate immunity developed only in the coelomates, after they split from pseudocoelomates and acoelomates. We hypothesize that the function of TLR-mediated immunity is to prevent microbial infection in the body cavity present only in the coelomates. Phylogenetic analysis showed that almost all arthropod TLRs form a separate cluster from the mammalian counterparts. We further hypothesize that TLR-mediated immunity developed independently in the protostomia and deuterostomia coelomates. PMID- 15114423 TI - Evidence for the existence of granzyme-like serine proteases in teleost cytotoxic cells. AB - Granzymes are granule-associated serine proteases, which are important effector molecules in NK cell and CTL functions. The granzyme family poses a perplexing problem in phylogenetics due to the lack of nonmammalian sequence information. We now report the identification of a cDNA that codes for a granzyme homologue, channel catfish granzyme-1 (CFGR-1), from nonspecific cytotoxic cells (NCC) of a teleost. NCC are the first identified and extensively studied cytotoxic cell population in teleosts. Ictalurus punctatus (channel catfish) granzyme cDNA encodes a protein with approximately 50% similarity to granzymes A and K. Highly conserved catalytic triad residues of serine proteases and other motifs common to granzymes were also identified. Conserved amino acid sequences, structure function data available for the serine protease family, and the crystal structure of human granzyme K supported a model of CFGR-1. It suggested an Arg/Lys primary substrate specificity that is shared with granzymes A and K. Furthermore, CFGR-1 has the four conserved disulfide bonds of granzymes A, K, and M. Phylogenetic analysis suggested that this molecule is a member of the granzyme family. Expression of CFGR-1 in NCC was confirmed by RT-PCR analysis. Presence of a granzyme-like molecule that might play an important role in the effector functions of NCC indicates that cell-mediated immunity with granule exocytosis and Fas pathways have been conserved for more than 300 million years. PMID- 15114424 TI - Concerted evolution and higher-order repeat structure of the 1.709 (satellite IV) family in bovids. AB - The 1.709 or satellite IV repeated DNA family originally isolated from the domestic cow was analyzed using Southern blotting, pulsed field gel electrophoresis, fluorescence in situ hybridization, and DNA sequencing in species belonging to the genera Bos, Bison, Bubalus, Syncerus, Boselaphus, and Tragelaphus. Hybridization indicates that the family has been amplified in Bos, Bison, Bubalus, and Syncerus but not in Boselaphus or Tragelaphus. Pericentromeric, higher-order repeat substructure exists in all species, with multimeric arrays ranging in size from 10 to 1500 kb. Sequence analysis of a 492 bp PCR product revealed comparable levels (0.2-4.5%) of intra- and interspecific divergence when species of Bos and Bison were compared, supporting the idea that species of these two genera should be recognized under the genus Bos. Alternatively, all Syncerus sequences cluster as a monophyletic group on an evolutionary tree and differ from those of Bos/ Bison by about 13%. Comparing these findings with the fossil record indicates that concerted evolution has occurred since Bos/ Bison and Syncerus last shared a common ancestor (5.0 MYA) but before the radiation of the genus Bos (2.5 MYA): GenBank accession numbers AY517856-AY517904. PMID- 15114425 TI - Purifying selection and demographic expansion affect sequence diversity of the ligand-binding domain of a glutamate-gated chloride channel gene of Haemonchus placei. AB - Ninety-five genomic sequences of the ligand-binding domain of glutamate-gated chloride channel genes of three populations of the parasitic nematode H. placei were evaluated for patterns of diversity, demography, and selection. These genes code for subunits of ion channels, which are involved in the mode of action of the most commonly used antiparasitic drugs, the macrocyclic lactones. An extremely high frequency of unique segregating sites in exons and introns was observed, with significantly negative neutrality tests in each population for noncoding, synonymous, and nonsynonymous sites. Several tests indicated that support for balancing selection, positive selection, and hitchhiking was lacking. McDonald-Kreitman tests using H. contortus or C. elegans as an outgroup revealed an extreme excess of replacement polymorphism, consistent with weak purifying selection. Although these tests agree that negative selection may explain the excess of replacement changes, an alternative interpretation is required for the significantly negative Fu and Li's D statistics based on silent and noncoding sites. These include homogeneous forces such as background selection and demographic expansion. The lack of population subdivision and the negative values of Tajima's D for this outbreeding parasitic nematode render background selection less likely than demographic expansion. Comparison of D statistics based on different site types using neutral coalescent simulations supported this interpretation. Although this statistic was more negative for nonsynonymous sites than for synonymous sites, most comparisons of the D statistic were not significantly different between mutation classes. A few significant site comparisons were also consistent with demographic expansion, because the observed test statistic ( D(neutral) - D(selected)) were low relative to the neutral expectations. Finally, previous mitochondrial studies also identified a demographic expansion of this parasitic nematode species, which lends further support to a scenario involving both demographic and purifying forces in the ligand-binding domain of H. placei. PMID- 15114426 TI - The deep phylogeny of land plants inferred from a full analysis of nucleotide base changes in terms of mutation and selection. AB - The occurrence frequencies of nucleotide bases are biased to those of T and A bases even at third codon positions for conserved amino acid residues with fourfold degeneracy in the chloroplasts of land plants. Regarding this bias as the result of selection, the base changes at these positions are fully analyzed theoretically in terms of mutation and selection. Although the degree of bias is considerably different depending on the lineages of land plants, the theoretical curves considering the influence of selection in the respective lineages provide a reasonable set of evolutionary distances for the relative base change probabilities estimated empirically from base changes enumerated in the comparison of rbcL genes. By using the fossil records of earliest seed plants in the Late Devonian and of uniaperturate and triaperturate pollen types in the early stage of the Cretaceous as calibration points, the divergence of Marchantiidae and a common ancestor of other land plants is estimated to have occurred 509 Mya, together with the estimation of a mutation rate of 1.45 x 10( 9) year(-1) per site. The other bryophytes such as Bryopsida, Anthocerotopsida, and Jungermanniidae are sister groups to tracheophytes, the divergence of bryophytes and tracheophytes being estimated to have occurred 483 Mya. The evolutionary distance of Gnetopsida from Coniferopsida and Magnoliophyta is concluded to be decisively longer than the distance between Coniferopsida and Magnoliophyta, i.e., the former divergence corresponds to 286 Mya and the latter to 211 Mya. PMID- 15114427 TI - Modern MRI tools for the characterization of acute demyelinating lesions: value of chemical shift and diffusion-weighted imaging. AB - Acute demyelinating lesions occur in various inflammatory disorders of the CNS. Apart from multiple sclerosis, most cases can be attributed to an overshooting immunological response to infectious agents called acute disseminated encephalomyelitis (ADEM). ADEM, which is mostly characterized by a monophasic course, has a multiphasic variant (MDEM). The early application of corticosteroids has been shown to be beneficial for the outcome; thus, an early diagnosis is highly desirable. Furthermore, the differential diagnosis ruling out neoplastic disorders may be difficult using conventional MRI alone. The potential diagnostic value of advanced MR techniques such as chemical shift imaging (CSI) and diffusion-weighted imaging (DWI) was investigated in a patient with MDEM, who had a new lesion in continuity with the initial disease manifestation. CSI was performed at 1.5 T with a long echo time of 135 ms for the evaluation of N-acetyl aspartate (NAA) and choline (Cho) and with short TE of 30 ms for macromolecules (mm) and myo-Inositol (mI). DWI was performed using a single-shot isotropic EPI sequence. Whereas acute and chronic areas of demyelination were neither distinguishable on T2- nor on contrast-enhanced T1-weighted images, CSI and DWI revealed different metabolite concentrations and diffusion characteristics within the composite lesion, clearly separating acute from chronic areas of demyelination. In conclusion, the addition of CSI and DWI may add to the diagnostic power of MRI in the setting of demyelinating disorders by identifying areas of acute and chronic demyelination, even in the absence of contrast enhancement. PMID- 15114428 TI - Detection and incidence of muscular adverse drug reactions: a prospective analysis from laboratory signals. AB - AIMS: The awareness of muscular adverse drug reactions (ADRs) increased since the withdrawal of cerivastatin, a HMG-CoA reductase inhibitor, from the market in August 2001. Our objectives were to assess the detection and incidence of muscular ADRs in a University Hospital using biochemical laboratory data and to evaluate the underreporting rate of drug-induced muscular disorders. METHODS: A prospective study was undertaken at Toulouse University Hospital, France, for 1 week per month from November 2001 to October 2002. Patients were selected by means of a computerized process using biochemical laboratory data based on serum creatine phosphokinase (CPK) values (over twofold normal). Medical records of all selected patients were then consulted. RESULTS: During the period of the study, 2017 CPK tests were performed, among which 171 values were over twofold normal corresponding to 129 patients. Because of lack of data, 26 patients were excluded. Among these patients ( n=103), 28 cases of muscular ADRs were suspected, 22 of which were detected in outpatient departments. Four patients were totally asymptomatic and five had an increase of CPK over fivefold normal. Nine cases were classified as "serious". Withdrawal of suspected drugs were done in 16 cases with regression of ADRs in 13 cases. According to hospitalization data, the incidence of muscular ADRs was estimated as 7.2 (2.6-15.7) per 10,000 inpatients and 9.3 (5.8-14.1) per 10,000 outpatients over 12 weeks. The involved drugs were mainly: statins (46.4%), fibrates (14.3%), antiretrovirals (14.3%), angiotensin-II receptor antagonists (10.7%), immunosuppressants (7.1%) or hydroxychloroquine (7.1). Only two cases, judged as "serious", were spontaneously reported by physicians during the same period. CONCLUSION: The results of this survey underline the importance to take into account drug hypothesis in muscular injuries diagnosis. PMID- 15114430 TI - Bioavailability of fluticasone propionate and mometasone furoate aqueous nasal sprays. AB - OBJECTIVES: To compare the systemic exposure for intranasal mometasone furoate (MF) and fluticasone propionate (FP) aqueous nasal sprays (ANS) in terms of serum and urinary cortisol parameters and plasma pharmacokinetics. METHODS: Twelve healthy subjects completed this three-way, cross-over study. They received FPANS (50 microg/spray), MFANS (50 microg/spray) or placebo ANS, eight sprays per nostril every 8 h for 4 days. Cortisol measurements were made at baseline and day 4. FP and MF plasma concentrations were also measured on day 4. RESULTS: MFANS produced similar mean plasma AUC (123 pmol/l h) to FPANS (112 pmol/l h). Despite the use of high doses, necessary to generate adequate pharmacokinetic data, only minor reductions in cortisol parameters were found, with no difference between FPANS and MFANS. CONCLUSIONS: FP and MF have similar and very low systemic bioavailability when administered intranasally using a high-dose regimen. It is therefore unlikely that therapeutic doses of intranasal FP or MF will produce dissimilar or significant degrees of systemic exposure or systemic effects. PMID- 15114429 TI - Oral administration of a low dose of midazolam (75 microg) as an in vivo probe for CYP3A activity. AB - OBJECTIVE: We investigated whether the oral administration of a low dose (75 micro g) of midazolam, a CYP3A probe, can be used to measure the in vivo CYP3A activity. METHODS: Plasma concentrations of midazolam, 1'OH-midazolam and 4'OH midazolam were measured after the oral administration of 7.5 mg and 75 micro g midazolam in 13 healthy subjects without medication, in four subjects pretreated for 2 days with ketoconazole (200 mg b.i.d.), a CYP3A inhibitor, and in four subjects pretreated for 4 days with rifampicin (450 mg q.d.), a CYP3A inducer. RESULTS: After oral administration of 75 micro g midazolam, the 30-min total (unconjugated + conjugated) 1'OH-midazolam/midazolam ratios measured in the groups without co-medication, with ketoconazole and with rifampicin were (mean+/ SD): 6.23+/-2.61, 0.79+/-0.39 and 56.1+/-12.4, respectively. No side effects were reported by the subjects taking this low dose of midazolam. Good correlations were observed between the 30-min total 1'OH-midazolam/midazolam ratio and midazolam clearance in the group without co-medication (r(2)=0.64, P<0.001) and in the three groups taken together (r(2)=0.91, P<0.0001). Good correlations were also observed between midazolam plasma levels and midazolam clearance, measured between 1.5 h and 4 h. CONCLUSION: A low oral dose of midazolam can be used to phenotype CYP3A, either by the determination of total 1'OH-midazolam/midazolam ratios at 30 min or by the determination of midazolam plasma levels between 1.5 h and 4 h after its administration. PMID- 15114431 TI - Pharmacokinetics of midazolam in CYP3A4- and CYP3A5-genotyped subjects. AB - OBJECTIVE: We investigated whether differences in pharmacokinetics of midazolam, a CYP3A probe, could be demonstrated between subjects with different CYP3A4 and CYP3A5 genotypes. METHODS: Plasma concentrations of midazolam, and of total (conjugated + unconjugated) 1'OH-midazolam, and 4'OH-midazolam were measured after the oral administration of 7.5 mg or of 75 micro g of midazolam in 21 healthy subjects. RESULTS: CYP3A5*7, CYP3A4*1E, CYP3A4*2, CYP3A4*4, CYP3A4*5, CYP3A4*6, CYP3A4*8, CYP3A4*11, CYP3A4*12, CYP3A4*13, CYP3A4*17 and CYP3A4*18 alleles were not identified in the 21 subjects. CYP3A5*3, CYP3A5*6, CYP3A4*1B and CYP3A4*1F alleles were identified in 20, 1, 4 and 2 subjects, respectively. No statistically significant differences were observed for the AUC(inf) values between the different genotypes after the 75- micro g or the 7.5-mg dose. CONCLUSION: Presently, CYP3A4 and CYP3A5 genotyping methods do not sufficiently reflect the inter-individual variability of CYP3A activity. PMID- 15114432 TI - Neuroendocrinological and neuropsychological correlates of dopaminergic function in nicotine dependence. AB - RATIONALE: There is multiple evidence that nicotine--as with ethanol and other drugs of abuse--stimulates dopamine release in the ventral striatum as a central part of the brain reward circuits. Chronic nicotine exposure leads to changes in these dopaminergic reward circuits. During nicotine withdrawal, an impaired dopaminergic function has been reported. On the behavioral level, this seems to result in motivational disturbances in abstaining smokers. OBJECTIVES: To investigate the impact of smoking on dopaminergic function in humans both on a neuroendocrinological and on a neuropsychological level. METHODS: Thirty-seven healthy smokers were assessed whilst smoking (test 1) and after abstaining overnight for 12 h (test 2). A control group of 18 non-smokers was also examined twice. Severity of nicotine dependence, incentive motivation, digit span and verbal fluency were assessed. The sensitivity of central dopamine (DA) D2 receptors was assessed with the apomorphine-induced growth hormone (GH) secretion. RESULTS: ANOVA revealed that GH response was significantly lower in smokers than in non-smokers (P=0.04). The GH response was significantly inversely correlated with severity of nicotine dependence (r=-0.39). Neuropsychological performance was not influenced by smoking status. After overnight abstinence from nicotine GH response, digit span and verbal fluency were not affected, whereas incentive motivation was significantly impaired in smokers (P=0.04). CONCLUSIONS: Smoking is significantly associated with a reduced sensitivity of central DA D2 receptors. This alteration of dopaminergic sensitivity is stable even after 12 h of abstinence from nicotine. Therefore, the hypothesis that the motivational impairment during withdrawal from nicotine is associated with an altered sensitivity of central DA D2 receptors cannot be supported. PMID- 15114433 TI - Role of basolateral amygdala dopamine in modulating prepulse inhibition and latent inhibition in the rat. AB - RATIONALE: The dopamine (DA) projection to the basolateral amygdala (BLA) modulates nucleus accumbens (NAc) and medial prefrontal cortex (mPFC) DA transmission. Given the involvement of the BLA, and of NAc and mPFC DA, in select forms of information processing, we sought to determine the role of BLA DA in modulating prepulse inhibition (PPI) and latent inhibition (LI). OBJECTIVE: The effects of BLA D1 (SCH 23390) and D2/D3 (raclopride) receptor blockade on PPI and LI were examined. METHODS: Separate groups of male Long-Evans rats received bilateral intra-BLA infusions of SCH 23390 (3.2 or 6.4 microg/0.5 microl per side), raclopride (2.5 or 5.0 microg/0.5 microl per side) or saline prior to testing. In two experiments, the effects of BLA DA receptor antagonism on PPI of the acoustic startle response (ASR) and LI of conditioned taste aversion were determined. A control group received bilateral intra-striatal infusions of SCH 23390 or raclopride prior to PPI testing. RESULTS: Intra-BLA SCH 23390 or raclopride had no effect on the ASR. Intra-BLA SCH 23390 enhanced and raclopride disrupted PPI, both in a dose-related manner. Intra-striatal SCH 23390 or raclopride had no effect on PPI or ASR magnitude. Finally, BLA DA receptor blockade had no effect on LI. CONCLUSIONS: These results indicate that PPI is modulated by BLA DA and suggest that this modulation occurs independently of changes in NAc and/or mPFC DA transmission. They also suggest that BLA DA is not involved in modulating LI and add to evidence indicating that PPI and LI are mediated by different neural substrates. PMID- 15114434 TI - Self-administration of fentanyl, cocaine and ketamine: effects on the pituitary adrenal axis in rhesus monkeys. AB - RATIONALE: Drugs of abuse can affect the functioning of the hypothalamic pituitary-adrenal (HPA) axis. Acute administration of drugs that serve as reinforcers have been observed to stimulate the rat HPA axis, leading to the suggestion that these stimulatory effects may contribute to the development of drug-maintained behaviors. OBJECTIVES: To determine whether reinforcing drugs that are dissimilar with respect to their mechanisms of action have similar effects on HPA axis activity at doses that are self-administered. Rhesus monkeys were randomly assigned to self-administer the mu-opioid agonist fentanyl, the psychomotor stimulant cocaine, or the NMDA antagonist ketamine. METHODS: Each monkey was trained to press a lever in order to receive an intravenous injection of drug or saline. Blood samples were obtained before, during, and after the self administration sessions and assayed for ACTH and cortisol by radioimmunoassay. RESULTS: Fentanyl, cocaine, and ketamine were each self-administered across a range of doses. However, the three drugs differed in their effects on ACTH and cortisol. Cocaine stimulated ACTH and cortisol secretion, a finding that is consistent with previous rat and primate studies. Self-administration of both fentanyl and ketamine inhibited HPA axis activity. HPA inhibition by fentanyl is consistent with other monkey and human studies, and contrasts with the stimulatory effects of mu-opioids in rodents. The inhibitory effect of ketamine on ACTH and cortisol secretion contrasts with findings in the few primate studies that have evaluated NMDA antagonists. Neither fentanyl nor cocaine, at doses that maintained maximum rates of responding, produced significant changes in ACTH and cortisol levels. CONCLUSIONS: There appears to be little commonality between different classes of abused drugs and their effects on the HPA axis, which calls into question the necessity for HPA axis stimulation in the reinforcement of drug maintained behavior. PMID- 15114436 TI - Effect of bisphenol A on drug metabolising enzymes in rat hepatic microsomes and precision-cut rat liver slices. AB - In order to assess the effects of bisphenol A (BPA) on enzymes of phase I and II biotransformation, studies were conducted in hepatic microsomes and precision-cut liver slices from male Sprague-Dawley rats. A testosterone hydroxylation assay was used for probing the activity of cytochrome P450 (CYP) forms, and an appropriate HPLC method for the separation of testosterone metabolites was developed. BPA markedly inhibited the hydroxylation of testosterone at 2alpha and 16alpha but not at 6beta or 7alpha, suggesting a differential inhibition of some CYP forms, in particular CYP2C11. This inhibitory effect was also observed when slices were first exposed to BPA and then incubated with testosterone in the absence of BPA, indicative of an irreversible inhibition of CYP. In liver slices, a differential conjugation of hydroxylated testosterone metabolites was observed, which was significantly decreased in the presence of BPA. BPA also inhibited the conjugation of the model compound umbelliferone. Pretreatment with BPA did not affect the conjugation of testosterone and umbelliferone. No hydroxylation, but extensive conjugation of BPA was observed upon incubation of liver slices with BPA alone or with testosterone or umbelliferone. The rapid and preferred conjugation, however, does not prevent the irreversible inhibition of some CYP forms by BPA. In conclusion, this study has shown that BPA causes a selective and irreversible inhibition of certain CYP forms and interferes with the conjugation of other drugs. PMID- 15114435 TI - Impaired set-shifting and dissociable effects on tests of spatial working memory following the dopamine D2 receptor antagonist sulpiride in human volunteers. AB - RATIONALE: Dopamine (DA) D(2) receptor antagonists have been shown to produce similar impairments to those seen in Parkinson's disease. These include working memory and set-shifting deficits. Theories of DA function have predicted that distraction or impaired switching may be important determinants of such deficits. OBJECTIVES: In order to test these hypotheses, we have followed up our previous findings with more refined tests (1) that allow measurement of spatial working memory (SWM) and distraction, (2) that allow separation of executive and mnemonic components of SWM and (3) that allow isolation of set-shifting from learning deficits. METHODS: Thirty-six young healthy male volunteers were tested on two occasions after oral administration of either 400 mg sulpiride or placebo. All participants performed the delayed response task. Sixteen participants received task-irrelevant distractors during this task, and were also given a self-ordered SWM test. The remaining participants were given delayed response tasks with task relevant distractors, and tests of attentional and task set-shifting. RESULTS: Sulpiride impaired performance of the delayed-response task both without distraction and with task-relevant distraction. By contrast, the drug protected against deficits from task-irrelevant distraction seen in the placebo group. Task set-switching was also impaired by sulpiride, with participants being slower to respond on switch trials compared with non-switch trials. There was also a trend for attentional set-shifting to be impaired following sulpiride. In contrast, self-ordered SWM performance was enhanced by sulpiride on the second test session only. CONCLUSIONS: These results support models of central DA function that postulate a role in switching behaviour, and in certain aspects of working memory. PMID- 15114437 TI - Interaction between uranium and the cytochrome c3 of Desulfovibrio desulfuricans strain G20. AB - Cytochrome c(3) of Desulfovibrio desulfuricans strain G20 is an electron carrier for uranium (VI) reduction. When D. desulfuricans G20 was grown in medium containing a non-lethal concentration of uranyl acetate (1 mM), the rate at which the cells reduced U(VI) was decreased compared to cells grown in the absence of uranium. Western analysis did not detect cytochrome c(3) in periplasmic extracts from cells grown in the presence of uranium. The expression of this predominant tetraheme cytochrome was not detectably altered by uranium during growth of the cells as monitored through a translational fusion of the gene encoding cytochrome c(3) ( cycA) to lacZ. Instead, cytochrome c(3) protein was found tightly associated with insoluble U(IV), uraninite, after the periplasmic contents of cells were harvested by a pH shift. The association of cytochrome c(3) with U(IV) was interpreted to be non-specific, since pure cytochrome c(3) adsorbed to other insoluble metal oxides, including cupric oxide (CuO), ferric oxide (Fe(2)O(3)), and commercially available U(IV) oxide. PMID- 15114438 TI - Hepatic haemangioendothelioma in adults: excellent outcome following liver transplantation. AB - Hepatic epithelioid haemangioendotheliomas (HEHEs) are rare, low-grade vascular tumours. Five adults with HEHEs and one adult with a vascular tumour showing combined features of haemangioma and haemangioendothelioma underwent liver transplantation. Two HEHE patients had extrahepatic metastases at the time of transplantation. Median survival time following diagnosis was 10.7 years (range 40 months to 195 months). One patient needed resection of a HEHE in the breast 13 years post-transplantation. All six patients are surviving free from disease 22 to 166 months after transplantation (median 77 months). One HEHE-patient who had been treated for 8 years for vertebral and cerebral localisations is free of disease without immunosuppression 56 months after transplantation. We can conclude that liver transplantation is a valuable treatment for hepatic haemangioendothelioma, even in cases of extrahepatic localisation of the disease. PMID- 15114439 TI - Clearance of C4d deposition after successful treatment of acute humoral rejection in follow-up biopsies: a report of three cases. AB - Acute humoral rejection (AHR) is currently perceived as an immunological reaction against donor antigens mediated by complement-binding antibodies. C4d, a split product of complement activation and bound to endothelial cells of the peritubular capillaries, is used as a diagnostic marker for AHR. We report on three patients with biopsy-proven acute humoral rejection who were treated initially with plasmapheresis (PS). As two of the patients did not recover renal function, and biopsy showed persistent C4d staining after PS, immunoadsorption (IAS) was additionally performed on them. In all patients, renal function recovered, and follow-up biopsies in two patients showed complete disappearance of C4d, 29 days and 58 days after transplantation and only minimal residual C4d deposits in one patient 48 days after transplantation. We conclude that successful treatment of AHR is followed by complete resolution of serological and histological markers of AHR, displayed by the disappearance of C4d. PMID- 15114440 TI - Intraspecific variation of mercury contamination in chicks of black-winged stilt (Himantopus himantopus) in coastal wetlands from southwestern Europe. PMID- 15114441 TI - Mercury distribution in farmlands downstream from an acetaldehyde producing chemical company in Qingzhen City, Guizhou, People's Republic of China. PMID- 15114442 TI - Distribution and concentration of trace metals in tissues of three penaeid shrimp species from Altata-Ensenada del Pabellon Lagoon (s.e. Gulf of California). PMID- 15114443 TI - Heavy metals in tissues of gray whales Eschrichtius robustus, and in sediments of Ojo de Liebre Lagoon in Mexico. PMID- 15114444 TI - Bioaccumulation of copper from contaminated wastewater by using Lemna minor. PMID- 15114445 TI - Chemical and ecotoxicological characterization of Averno Lake. PMID- 15114446 TI - Impact of acid mine drainage from the abandoned Halikoy mercury mine (western Turkey) on surface and groundwaters. PMID- 15114447 TI - Chlorinated hydrocarbons in skin and blubber of two blue whales (Balaenoptera musculus) stranded along the Baja California coast. PMID- 15114448 TI - Organochlorine pesticide residues in water, sediment, and muscle of river shad, Hilsa ilisha (Hamilton 1822) from the South Patches of the Bay of Bengal. PMID- 15114449 TI - Studies on biodegradation of 2,4-D and metribuzin in soil under controlled conditions. PMID- 15114451 TI - Levels and trends of chlorinated pesticides in human breast milk from Ankara residents: comparison of concentrations in 1984 and 2002. PMID- 15114450 TI - PAH contamination sources in the sediments of the Strait of Istanbul (Bosphorus), Turkey. PMID- 15114452 TI - Trace element contents in human head hair of residents from Agra City, India. PMID- 15114453 TI - Elevated exposure to nitrogen dioxide during food preparation: results from a cooking laboratory. PMID- 15114454 TI - Asbestos abatement of pipe and floor tile/mastic and comparison of critical plastic barrier controls. PMID- 15114455 TI - Temporal evolution of natural and man-made radioactivity levels in milk samples: dosimetry implications. PMID- 15114456 TI - Effects of funnel and gate geometry on capture of contaminated groundwater. PMID- 15114457 TI - Assessing stream grazer response to stress: a post-exposure feeding bioassay using the freshwater snail Lymnaea peregra (Muller). PMID- 15114458 TI - Use of the IC50 for predicting joint toxic effects of mixtures. PMID- 15114459 TI - Effects of body size and sodium chloride on the tolerance of net-spinning caddisfly larvae to fluoride toxicity. PMID- 15114460 TI - Levels of serum cholinesterase activity in the rococo toad (Bufo paracnemis) in agrosystems of Argentina. PMID- 15114461 TI - Toxicity of malathion and carbaryl pesticides: effects on some biochemical profiles of the freshwater fish Colisa fasciatus. PMID- 15114462 TI - Effects of pesticides and their hydrolysates on catalase activity in soil. PMID- 15114463 TI - Comet assay assessment of wastewater genotoxicity using yeast cells. PMID- 15114464 TI - Bioremediation of arsenic contaminated groundwater by modified mycelial pellets of Aspergillus fumigatus. PMID- 15114465 TI - Effects of genistein on growth and development of aquatic vertebrates. PMID- 15114466 TI - Acute toxicity of Bunker C refined oil to the Japanese littleneck clam Ruditapes philippinarum (Bivalvia: Veneridae). PMID- 15114467 TI - Effects of aluminum in acidic water on hematological and physiological parameters of the neotropical fish Leporinus macrocephalus (Anostomidae). PMID- 15114468 TI - Toxicity of four surfactants to juvenile rainbow trout: implications for use over water. PMID- 15114469 TI - Temporal changes in prevalence of diabetes and impaired glucose tolerance associated with lifestyle transition occurring in the rural population in India. AB - AIMS/HYPOTHESIS: The rural Indian population is undergoing lifestyle transition due to socio-economic growth. This study was done to determine the temporal changes in prevalence of diabetes and IGT that could have occurred in a rural population in India as a result of the lifestyle transition. METHODS: A cross sectional study of 1213 Asian-Indian subjects aged 20 years or over was done to look for the prevalence of diabetes and IGT using the 1999 WHO criteria. The temporal changes were assessed in comparison with a similar study conducted 14 years previously. The factors associated with the temporal changes were also analysed. RESULTS: Nearly a three-fold increase in age- and sex-adjusted prevalence of diabetes (from 2.20% to 6.36%) was seen in 2003 when compared with a similar study done 14 years before. Prevalence of IGT did not change significantly (7.44% in 1989 vs 7.18% in 2003). Improvement in living conditions had occurred during the period, occupational changes were seen, the number of manual labourers had decreased and economic conditions had improved. BMI and waist circumference had increased. After correcting for age, sex and differences in time periods, waist circumference and physical inactivity showed significant associations with the increased prevalence of diabetes. CONCLUSIONS/INTERPRETATION: Demographic transition due to improved living conditions in rural India was associated with a three-fold increase in the prevalence of diabetes. Increased upper body adiposity and physical inactivity showed significant association with this phenomenon. PMID- 15114470 TI - Parallel manifestation of insulin resistance and beta cell decompensation is compatible with a common defect in Type 2 diabetes. AB - AIMS/HYPOTHESIS: The aim of the study was to evaluate the relationship between insulin sensitivity, beta cell function and glucose tolerance, and its dependence on variants in the newly identified Type 2 diabetes susceptibility gene, calpain 10 ( CAPN10). METHODS: We studied 203 men of the same age but with varying degrees of glucose tolerance. These men participated in (i) an oral glucose tolerance test, (ii) a euglycaemic clamp combined with indirect calorimetry and infusion of [3-(3)H]-glucose and (iii) a stepwise assessment of acute insulin response to arginine (AIR) at three different glucose concentrations (fasting, 14 and 28 mmol/l). RESULTS: There was a linear increase in NEFA levels ( p<0.0005) and WHR ( p<0.0005) and decrease in glucose uptake due to a reduction in glucose storage over the entire range of glucose tolerance ( r=-0.404; p<0.005). No increase in endogenous glucose production (EGP) was seen until patients had manifest diabetes. However, when EGP was expressed relative to fasting insulin concentrations, there was a linear deterioration of basal hepatic insulin sensitivity ( r=-0.514; p<0.005). The AIR followed a bell-shaped curve with an initial rise and subsequent decrease. However, AIR adjusted for insulin sensitivity (disposition index) showed a linear decrease with increasing glucose concentrations ( r=-0.563; p<0.001) starting already in subjects with normal glucose tolerance. There was an inverse correlation between increase in WHR and NEFA and peripheral as well as hepatic insulin sensitivity. Subjects with the genotype combination of CAPN10 consisting of SNP44 TT and SNP43 GG genotypes had significantly lower insulin-stimulated glucose uptake than carriers of the other genotype combinations (5.3+/-0.4 vs 7.2+/-0.4 mg.ffm kg(-1).min(-1).mU.l(-1); p<0.005). CONCLUSIONS/INTERPRETATION: We conclude that the pre-diabetic state is characterised by a similar linear deterioration of peripheral and hepatic insulin sensitivity as beta cell function and that variants in the CAPN10 gene modify this relationship. These findings are compatible with a common defect in muscle, liver and beta cells in the pathogenesis of Type 2 diabetes. PMID- 15114472 TI - The Rxo1/ Rba1 locus of maize controls resistance reactions to pathogenic and non host bacteria. AB - Infiltration of different maize lines with a variety of bacterial pathogens of maize, rice and sorghum identified qualitative differences in resistant reactions. Isolates from two bacterial species induced rapid hypersensitive reactions (HR) in some maize lines, but not others. All isolates of the non-host pathogen Xanthomonas oryzae pv. oryzicola (bacterial leaf streak disease of rice) and some isolates of the pathogenic bacterium Burkholderia andropogonis induced HR when infiltrated into maize line B73, but not Mo17. Genetic control of the HR to both bacteria segregated as a single dominant gene. Surprisingly, both phenotypes mapped to the same locus, indicating they are either tightly linked or controlled by the same gene. The locus maps on the short arm of maize chromosome six near several other disease-resistance genes. Results indicate the same type of genes may contribute to both non-host resistance and resistance to pathogens. PMID- 15114471 TI - Control of glycaemia: from molecules to men. Minkowski Lecture 2003. AB - Regulation of glycaemia represents a fundamental biological principle, and its failure underlies Type 2 diabetes. The complex aetiology of Type 2 diabetes, which probably involves a medley of molecular mechanisms, requires dissection out of diabetes-associated subphenotypes, such as the non-obese with increased liver fat or the obese with low plasma adiponectin. The concepts of the hyperbolic relationship of insulin secretion and insulin sensitivity with glucose allostasis help us to establish the pathophysiological framework within which such mechanisms must operate. The translation of burgeoning new basic science findings into a physiological and clinical context calls for novel and imaginative clinical experimental tools. For the purpose of this review, four molecules (adiponectin [APM1], stearoyl CoA desaturase-1 [SCD1], insulin receptor substrate 1 [IRS1], peroxisome proliferator-activated receptor-gamma [PPARG]), each with a plausible role in the disease process, have been selected to illustrate the use of such techniques in humans. These include procedures as diverse as isotope dilution for turnover studies (e.g. glycerol turnover as a proxy for lipolysis), conventional and modified clamp procedures, association studies of functionally relevant single nucleotide polymorphisms in candidate genes (e.g. IRS-1 and PPAR gamma), multivariate correlational analyses (as with plasma adiponectin), magnetic resonance spectroscopy to quantify intra-tissue lipid deposition and regional fat distribution, and gas chromatography to determine fatty acid patterns in selected lipid fractions as proxy for intrahepatic enzyme activity. A concerted effort by scientists from many disciplines (genetics and cell biology, physiology and epidemiology) will be required to bridge the growing gap between basic scientific concepts of biological modifiers of glycaemia and concepts that are truly relevant for human Type 2 diabetes. PMID- 15114473 TI - Production and characterization of somatic hybrids between upland cotton (Gossypium hirsutum) and wild cotton (G. klotzschianum Anderss) via electrofusion. AB - Symmetric somatic hybrid plants between Gossypium hirsutum Coker 201 and G. klotzschianum were obtained through electrofusion. The fusion products were cultured in KM8P medium supplemented with 2.685 microM alpha-naphthaleneacetic acid and 0.465 microM kinetin, and the regenerated plants were morphologically, genetically, and cytologically characterized. Nuclear-DNA flow cytometric analysis revealed that the plants tested (31 of 40) had a relative DNA content close to the total DNA contents of the two parents. Subsequent genome DNA analysis using random amplified polymorphic DNA markers revealed 16 of 18 plants were true somatic hybrids. Cytological investigation of the metaphase root-tip cells of seven hybrids revealed there were 72-81 chromosomes in the hybrids, a value close to the expected 78 chromosomes. The morphology of the hybrids was distinct from that of the parents and from that of the regenerants from protoplasts of Coker 201. Somatic hybridization represents a potent and novel tool for transferring genomes of wild cottons containing economically important traits to cultivars in breeding programs. This is the first report on the regeneration of somatic hybrids via protoplast fusion in cotton. PMID- 15114474 TI - Physical mapping and putative candidate gene identification of a quantitative trait locus Ctb1 for cold tolerance at the booting stage of rice. AB - Norin-PL8 is a cold-tolerant variety of rice (Oryza sativa L.) that was developed by introgressing chromosomal segments from a cold-tolerant tropical japonica variety, Silewah, into a template japonica variety, Hokkai241. We previously identified two closely linked quantitative trait loci, Ctb1 and Ctb2, for cold tolerance at the booting stage of Norin-PL8 in the long arm of chromosome 4. We report here the physical mapping of Ctb1 and the identification of the candidate genes. A total of 2,008 segregating individuals were screened for recombination in the Ctb1 region by a PCR-based screening, and a series of near-isogenic lines (NILs) were developed from progenies of recombinants. A comparison of the degrees of cold tolerance of the NILs indicated that Ctb1 is located in the 56-kb region covered by a bacterial artificial chromosome clone, OSJNBa0058 K23, that had been sequenced by the International Rice Genome Sequence Project. We found seven open reading frames (ORFs) in the 56-kb region. Two ORFs encoded receptor-like protein kinases that are possibly involved in signal transduction pathways. Proteins that may be associated with a ubiquitin-proteasome pathway were encoded by three ORFs, two of which encoded F-box proteins and one of which encoded a protein with a BAG domain. The other two ORFs encoded a protein with an OTU domain and an unknown protein. We were also able to show that Ctb1 is likely to be associated with anther length, which is one of major factors in cold tolerance at the booting stage. PMID- 15114475 TI - [Radiological diagnostics of Hodgkin- and non-Hodgkin lymphomas of the thorax]. AB - Malignant lymphomas belong to the most important malignant diseases in western countries with an increasing incidence of Non-Hodgkin lymphoma. The thorax is the location of primary manifestation especially in patients with Hodgkin's disease. Progression of disease and therapy associated complications are frequently located in the chest. Based on morphological imaging criteria the two types of lymphoma cannot be differentiated, helpful for differentiation is, however, the way of disease spread. Primary and secondary thoracic lymphoma represent a diagnostic challenge in radiology: the patterns are variable in radiography as well as in computed tomography and alter under therapy. Radiological studies, especially CT, are an integral part of the staging process. MRI is considered advantageous for chest wall disease. PET as functional imaging technique has its proven role for staging of high grade lymphomas, the combination of functional and morphological information provided by PET-CT will become the first diagnostic standard in the future. PMID- 15114476 TI - [The reconstruction of scaphoid pseudoarthroses with the operation of Matti Russe. A retrospective follow-up analysis of 84 patients]. AB - BACKGROUND: In a retrospective study 84 patients were examined who were treated with a Matti-Russe procedure between 1985 and 1997 due to scaphoid non-union. The purpose of this study was to evaluate the long term results after scaphoid reconstruction and to get impressions about subjective feeling of the patients. PATIENTS AND METHODS: 79 patients were male, 5 were female. The average follow-up period was 88 months. Measured parameters were: grip strength and range of motion, pain was evaluated with a visual analog scale (VAS) from 0 to 100. The functional and subjective outcome was evaluated with the DASH-questionnaire. RESULTS: A bony consolidation could be verified in 82% of the patients. The mean postoperative pain score was 3 (non-stress) and 33 (stress) in patients with scaphoid union. The DASH-score reached 15. Active range of motion and grip strength were 82% and 92% compared to the contralateral side. 81% of the patients have been working in strenuous jobs. CONCLUSION: The results show the reliability of the Matti-Russe procedure in non-union of fractures of the scaphoid. Alternative treatment options have no advantages in bony union. PMID- 15114477 TI - Reconciliation and post-conflict third-party affiliation among wild chimpanzees in the Mahale Mountains, Tanzania. AB - This study investigated post-conflict (PC) behavior among wild chimpanzees (Pan troglodytes) of the M-group in the Mahale Mountains, Tanzania, and examined what types of behavior characterize the PC situation in this group, and the factors that influence the occurrence of PC affiliation between opponents soon after the end of an aggressive conflict (i.e., reconciliation). We found that the opponents affiliated selectively soon after the end of aggression, suggesting that reconciliation occurred in this group. The mean individual corrected conciliatory tendency (CCT) (Veenema et al. 1994 in Behav Proc 31:29-38) was 14.4%, which is similar to or lower than frequencies observed in studies of captive and wild chimpanzees. The valuable relationship hypothesis predicts that the CCT is higher among individuals who share valuable relationships (e.g., males or affiliative dyads) than among individuals who do not (e.g., females or less-associative dyads). However, the analysis based on data for aggression between unrelated individuals (including one incident between an adult and non-adult) and aggression between unrelated adults, did not uncover this difference. Affiliation by a previously uninvolved individual with the victim ("consolation") and with the aggressor ("appeasement") occurred more frequently following aggression than in the control condition. The results are compared with previous studies of captive and wild chimpanzees. PMID- 15114478 TI - Dislocations with intervertebral disc prosthesis: two case reports. AB - To date, only three cases of artificial disc prosthesis dislocation have been reported in the literature. We present in detail two additional cases of prosthesis dislocation and discuss the surgical interventions undertaken that resulted in a good clinical outcome in both patients. PMID- 15114479 TI - Catastrophic heparin-induced thrombocytopenia/thrombosis syndrome related to the use of a Port-A-Cath in a breast cancer patient receiving chemotherapy. AB - Heparin-induced thrombocytopenia and thrombosis (HIT/T) syndrome is usually triggered by an immune response after repeated administration of heparin. The syndrome is strongly associated with limb deep vein thrombosis and is potentially life-threatening if unrecognized. We describe the case of a patient with compartment syndrome of the left forearm complicated by HIT/T that developed after Port-A-Cath implantation through the left subclavian vein. Prompt recognition of HIT/T, immediate withdrawal of heparin, and timely institution of thrombolytic therapy successfully prevented limb loss. PMID- 15114481 TI - The proteome expression patterns in adult Ascaris suum under exposure to aerobic/anaerobic environments analyzed by two-dimensional electrophoresis. AB - We investigated the protein expression patterns (the proteome) in adult Ascaris suum under exposure to aerobic and anaerobic environments in vitro using two dimensional polyacrylamide gel electrophoresis. Adult A. suum were cultured in RPMI-1640 in the absence of oxygen (anaerobic) or in its presence (aerobic), and whole worm extracts and their excretory-secretory (ES) products were prepared for protein analysis. More than 200 protein spots resolved in two-dimensional gels. A comparison of aerobic versus anaerobic cultures revealed two aerobic and three anaerobic specific spots differentially expressed in adult worm extracts. ES products released during aerobic/anaerobic cultures also exhibited one aerobic and seven anaerobic specific spots in two-dimensional maps. One of the anaerobic specific protein spot was analyzed for its internal amino acid sequence (1 LLAFELAPHGIR-12; 1-VNTVNPGAVD-10) and the protein was predicted to be a putative dehydrogenase. These findings suggest that the availability of oxygen or its absence in the environment may influence protein expression patterns in parasitic nematodes. PMID- 15114480 TI - A proteomic approach to understanding the development of multidrug-resistant Candida albicans strains. AB - Resistance of the pathogenic yeast Candida albicans to the antifungal agent fluconazole is often caused by the overexpression of genes that encode multidrug efflux pumps ( CDR1, CDR2, or MDR1). We have undertaken a proteomic approach to gain further insight into the regulatory network controlling efflux pump expression and drug resistance in C. albicans. Three pairs of matched fluconazole susceptible and resistant clinical C. albicans isolates, in which drug resistance correlated with stable activation of MDR1 or CDR1/2, were analyzed for differences in their protein expression profiles. In two independent, MDR1 overexpressing, strains, additional up-regulated proteins were identified, which are encoded by the YPR127 gene and several members of the IFD ( YPL088) gene family. All are putative aldo-keto reductases of unknown function. These proteins were not up-regulated in a fluconazole-resistant strain that overexpressed CDR1 and CDR2 but not MDR1, indicating that expression of the various efflux pumps of C. albicans is controlled by different regulatory networks. To investigate the possible role of YPR127 in the resistance phenotype of the clinical isolates, we constitutively overexpressed the gene in a C. albicans laboratory strain. In addition, the gene was deleted in a C. albicans laboratory strain and in one of the drug-resistant clinical isolates in which it was overexpressed. Neither forced overexpression nor deletion of YPR127 affected the susceptibility of the strains to drugs and other toxic substances, suggesting that the regulatory networks which control the expression of efflux pumps in C. albicans also control genes involved in cellular functions not related to drug resistance. PMID- 15114482 TI - Ultrastructure of eggshell of Chrysomya nigripes Aubertin (Diptera: Calliphoridae). AB - We examined the eggshell structure of the blowfly, Chrysomya nigripes Aubertin, a species of forensic importance, using scanning electron microscopy. A relatively wide plastron region was located dorsally. It extended almost the entire length of the egg and bifurcated slightly, close to the micropyle, which is a deep cavity surrounded by an elevated, fine, wavy rim. The chorionic sculpture had an hexagonal pattern, with a smooth and elevated boundary. The ruptured eggshell showed the outermost exochorion, membranous outer endochorion and pillar layer, which also had an hexagonal pattern on a perforated mesh pillar layer. This study provides a greater database on the egg of this forensically significant blowfly. PMID- 15114484 TI - Expression of integrin subunits alphav and beta3 in acute lung inflammation. AB - Integrin subunits alphav and beta3 form a dimer, alphavbeta3, which is expressed on normal neutrophils and endothelium. We investigated the expression of integrin subunits alphav and beta3 in acute lung inflammation in Sprague-Dawley rats ( n=5 each) following intratracheal challenge with Escherichia coli or Streptococcus pneumoniae, which induce neutrophil recruitment through different mechanisms. Control rats ( n=5) were given endotoxin-free saline. Both bacterial challenges induced similar levels of recruitment of neutrophils in lungs. Western blots showed lower expression of integrin subunits alphav and beta3 in lungs challenged with E. coli compared to those given S. pneumoniae. Immunohistochemistry and immunogold electron microscopy localized both integrin subunits in neutrophils and endothelium in the control and treated rat lungs. Quantitative immunohistochemistry showed that E. coli-challenged rat lungs contained a lower percentage of neutrophils expressing integrin subunits alphav and beta3 compared to those challenged with S. pneumoniae ( P<0.05). We conclude that E. coli infection decreased the percentage of neutrophils expressing integrin subunits alphav and beta3 compared to S. pneumoniae infection. These data lay the foundation for further characterization of these integrin subunits in neutrophil migration specifically in S. pneumoniae infection that utilizes molecules other than beta2 integrins for neutrophil recruitment. PMID- 15114485 TI - Studies on seasonal arthropod succession on carrion in the southeastern Iberian Peninsula. AB - A global study of the sarcosaprophagous community that occurs in the southeastern Iberian Peninsula during all four seasons is made for the first time, and its diversity is described with reference to biological indices. A total of 18,179 adults and, additionally, a number of preimaginal states were collected. The results for the main arthropod groups, and their diversity are discussed in relation to the season and decompositional stages. The results provide an extensive inventory of carrion-associated arthropods. An association between decomposition stages and more representative arthropod groups is established. With respect to the biological indices applied, Margalef's index shows that the diversity of the community increases as the state of decomposition advances, while Sorenson's quantitative index shows that the greatest similarities are between spring and summer on the one hand, and fall and winter, on the other. PMID- 15114483 TI - Differences in the uptake of iron from Fe(II) ascorbate and Fe(III) citrate by IEC-6 cells and the involvement of ferroportin/IREG-1/MTP-1/SLC40A1. AB - Dietary iron is present in the intestine as Fe(II) and Fe(III). Since enterocytes take up Fe(II) by the divalent metal transporter (DMT1), Fe(III) must be reduced. Whether other Fe(III) transport processes are present is unknown. Release of iron from the enterocyte into the plasma involves the iron-regulated transporter 1/metal transporter protein-1 (IREG-1/MTP-1, ferroportin) but ferroportin is also found on the apical membrane. We compared the uptake of iron from Fe(II):ascorbate and Fe(III):citrate using the rat intestinal enterocyte cell line-6 (IEC-6), in the presence of ferrous chelators, a blocking antibody to ferroportin, at different pH and during the over-expression of DMT1. Firstly, surface ferrireduction was absent. Secondly, blocking ferroportin partly and totally reduced Fe(II) and Fe(III) uptake, respectively. Thirdly, optimal Fe(II) uptake occurred at pH 5.5 but Fe(III) uptake was unaffected by pH and, fourthly, over-expression of DMT1 increased uptake of Fe(II) and Fe(III). This indicates that an increased extracellular H+ concentration facilitates DMT1-mediated Fe(II) uptake at the cell membrane. However, since Fe(III) uptake required DMT1, but not cell surface ferrireduction, and was independent of variations in extracellular pH, it appears that Fe(III) is internalised before ferrireduction and transport by DMT1. Ferroportin may function as a modulator of DMT1 activity and play a role in Fe(III) uptake, possibly by affecting the number or affinity of citrate binding sites. PMID- 15114486 TI - Entomofauna of buried bodies in northern France. AB - Autopsies of exhumed cadavers can reveal important evidence for clarification of medical insurance and social issues. This study concerns insects sampled on 22 exhumed cadavers in the Lille area. For each corpse, the species and the stages of development were noted, as well as the time elapsed after burial, the location of the cemetery, the stage of decay and possible preservation treatment. A total of eight Diptera and two Coleoptera species were sampled on the corpses. The relationships between entomofauna and conditions of burial are discussed. Three species were regularly found because of their preference for underground environments or closed environments: Conicera tibialis, typically associated with buried bodies, Leptocera caenosa which is known to be associated with human faeces, water closets, caves and cracked soil pipes, and Ophyra capensis, sometimes found on human bodies kept indoors for several months, where blowflies have not had access. Triphleba hyalinata, which is associated with human bodies in wooden coffins, was found only twice. PMID- 15114487 TI - Amyotrophic lateral sclerosis with neuronal intranuclear protein inclusions. AB - A 46-year-old patient developed amyotrophic lateral sclerosis (ALS) characterized by rapid progression. She needed respiratory assistance after a course of 9 months. She died 4.5 years after onset. Autopsy showed dramatic atrophy of the spinal cord, sparing only the posterior tracts, associated with neuronal loss and astrogliosis in various areas including the anterior horns, motor cortex, striatum, thalamus, and substantia nigra. Ubiquitin immunohistochemistry showed rare skein-like inclusions in the surviving spinal and medullary motor neurons. Eosinophilic inclusions were found in the nuclei of pyramidal neurons in the hippocampus. These inclusions were immunoreactive to antibodies against ubiquitin, promyelocytic leukemia gene product, proteasome, and ataxin-3. They were not immunoreactive to antibodies against tau, cystatin C, neurofilament, alpha-synuclein, SOD-1, and polyglutamine (1C2), and were not stained by ethidium bromide. Similar inclusions were found in the motor cortex. The immunoreactivity of the inclusions was similar to that encountered in diseases associated with CAG repeats, except for the negativity of the immunolabelling with 1C2. At the ultrastructural level, the nuclear inclusions were made of straight filaments (10 12 nm in diameter) arranged at random, reminiscent of the polyglutamine intranuclear hyaline inclusions. PMID- 15114488 TI - Wandering spleen: a rare cause of mesenteroaxial gastric volvulus. AB - Gastric volvulus is a rare cause of acute abdomen in children. Usually it is associated with defects such as diaphragmatic hernia, hiatal hernia, eventration, and paralysis of the diaphragm. We report an extremely rare case presenting with acute intractable vomiting and abdominal distention. Passage of a nasogastric tube relieved the symptoms. A history of the ingestion of a large bolus of chewing gum and a suboptimal contrast study were misleading, and there was a delay in diagnosis of 3 days. Later, repeat upper GI fluoroscopy with contrast medium identified mesenteroaxial volvulus of the stomach. Exploratory laparotomy additionally revealed a wandering spleen. Derotation of the stomach with anterior gastropexy was performed. To our knowledge, wandering spleen in association with gastric volvulus has been mentioned only three times before in the literature. PMID- 15114489 TI - Frictional properties of contacting surfaces in the hemelytra-hindwing locking mechanism in the bug Coreus marginatus (Heteroptera, Coreidae). AB - The structure and function of the hemelytra-to-hindwing locking mechanism of the bug Coreus marginatus were analysed. The system consists of a cuticular protrusion in the ventral side of the hemelytra, which locks the subcostal border of the hindwing in flight. The speed and distance slid by both surfaces against one another during flight were assessed using a combination of high-speed video recordings and a 2D geometrical model. The friction coefficient between sliding surfaces was assessed using a micromanipulator, coupled with force transducers. This was done under three experimental conditions: freshly dissected, air dried and rehydrated ethanol preserved samples. The results showed a high speed of sliding, approximately 0.18 m s(-1), with a relatively low friction coefficient (0.2 micro). There was no evident difference in the friction measured under the various treatments, with the exception of the rehydrated condition, which was lower. The surface morphology of the wing locking mechanism, namely outgrowths of one part having rounded edges, and completely flat surface on the counterpart, effectively aids in the reduction of friction at the microscopic level. The structure is effective even dry, and after being preserved in ethanol, suggesting that no cuticle secreted lubrication substance is responsible for its effectiveness. The ultrastructure presumably confers mechanical stability to the system under the high load it is subjected to in flight. PMID- 15114490 TI - Multislice spiral CT angiography in peripheral arterial occlusive disease: a valuable tool in detecting significant arterial lumen narrowing? AB - The aim of this study was to evaluate the potential of multislice CT angiography (CTA) in detecting hemodynamically significant (> or =70%) lesions of lower extremity inflow and runoff arteries. Fifty patients (42 men, 8 women; mean age 68 years) with peripheral arterial occlusive disease underwent multislice spiral CTA and digital subtraction angiography (DSA) from the infrarenal aorta to the supramalleolar region. CT parameters were 4x2.5-mm collimation, 15-mm table increment/rotation (pitch 6), and 1.25-mm reconstruction increment. Semitransparent volume rendering technique (STVR) images with semitransparent display of the arterial lumen (opacity: 50%) and vascular calcifications (opacity: 20%), as well as maximum intensity projection (MIP), and MIP together with axial CT studies were independently reviewed for hemodynamically significant lesions (> or =70% cross-sectional area reduction). DSA was the standard of reference. In 46 patients, 260 lesions were found (95 stenoses, 165 occlusions). For detecting > or =70% lesions in all vessel regions, sensitivity and specificity were 84% and 78% (STVR), 89% and 74% (MIP), and 92% and 83% (MIP+axial CT), respectively, with a significantly lower sensitivity of STVR ( p<0.05) and a significantly lower specificity of MIP studies ( p<0.01). Sensitivity and specificity were, respectively, 81% and 93% (STVR), 88% and 75% (MIP). and 92% and 95% (MIP+axial CT) at aortoiliac arteries, 92% and 73% (STVR), 95% and 70% (MIP) and 98% and 70% (MIP+axial CT) at femoropopliteal arteries, as well as 82% and 64% (STVR), 86% and 74% (MIP), and 90% and 74% (MIP+axial CT) at infrapopliteal arteries. Specificity of MIP-CTA was significantly lower in the aortoiliac region ( p<0.01), whereas STVR revealed significantly lower specificity at infrapopliteal arteries ( p<0.05). In the infrapopliteal region, the particular CTA imaging modalities led to misinterpretation regarding stenoses and occlusions in 39-45 cases, whereas only 0-6 significant aortoiliac and femoropopliteal lesions were misinterpreted. Multislice CTA is helpful in detecting hemodynamically significant lesions in peripheral arterial occlusive disease. Since axial CT studies yielded the most correct results, they should always be reviewed additionally. In the infrapopliteal region, exact lesion assessment remains problematic due to small vessel diameters. PMID- 15114491 TI - Renal dimensions measured by ultrasonography in children: variations as a function of the imaging plane and patient position. AB - The purpose of this study was to determine the effect of patient positioning on sonographic renal measurements and to test if the patient position alters the three-dimensional shape of the kidneys. The maximum longitudinal renal length and transverse renal width and depth were measured in the supine and prone position in 100 children (200 kidneys). Age ranged from 6 months to 16 years (mean age 5 years). The results were compared for statistically significant differences. The maximum measured longitudinal renal length was statistically significantly larger in the supine than in the prone position (supine position, left: 8.0 cm; right: 7.7 cm; prone position, left: 7.9 cm, right: 7.6 cm; P<0.001). There was no statistically significant change in the transverse diameters (width and depth, P>0.001) and renal volume ( P>0.001) in the supine vs. prone positions. Our results show that position-induced reshaping of the kidneys is unlikely to be responsible for the discrepancy in maximum longitudinal renal measurements comparing supine with prone positions. Position-dependent changes in the degree of filling of the renal calyces and pelvis as well as errors in caliper distance measurements for the different scan depths (supine vs. prone) are more likely to be responsible for the encountered differences. Consequently, we recommend to add prone renal length measurements in addition to the supine measurements. In follow up examinations, renal length measurements should only be compared that have been collected in the same patient position. PMID- 15114493 TI - Kava Hepatotoxicity: Are we any closer to the truth? AB - In recent years, kava kava ( Piper methysticum, Forst. f., Piperaceae) has been implicated in a number of liver failure cases. Ever since this has kept the scientific world busy. Even though, on closer inspection, the majority of the case reports are probably not connected to kava intake, hepatotoxic effects of kava cannot generally be ruled out. In this article the major theories as to the mechanism of kava hepatotoxicity are summarized. But in spite of all these hypotheses, there is still no satisfactory answer. In any case, further studies, that might hopefully restore the reputation of kava, are required. PMID- 15114492 TI - Agrobacterium tumefaciens-mediated transformation of Campanula carpatica: factors affecting transformation and regeneration of transgenic shoots. AB - An efficient transformation system for Campanula carpatica was developed using Agrobacterium tumefaciens strains LBA4404 (harbouring the plasmid pBI121), and AGL0 (harbouring the plasmid pBEO210). This is the first report on the transformation of C. carpatica. Various factors affecting the transformation efficiency and subsequent regeneration were identified. The age of seedlings from which the explants for transformation studies were taken, and the growth conditions under which the seedlings were grown had a significant influence on the production of transformed shoots. Hypocotyls taken from 12-day-old seedlings grown in the dark were the most productive, with up to 25% of hypocotyls producing transformed shoots. Explants taken from 5-week-old seedlings produced only transformed callus. The medium used for co-cultivation and incubation also had a significant influence on transformation frequency and shoot regeneration. The cultivar "Blue Uniform" was more responsive than "White Uniform". Both bacterial strains and plasmids were equally effective in producing transformed tissue. Transformed shoots were selected on kanamycin medium, and the presence of the uidA and nptII genes in those selected shoots was confirmed by beta glucuronidase and ELISA analyses, respectively. PMID- 15114495 TI - Novel diterpenoid acetylcholinesterase inhibitors from Salvia miltiorhiza. AB - Acetylcholinesterase (AChE, EC 3.1.1.7) inhibitors are the only registered drugs used to treat Alzheimer's disease (AD). New AChE inhibitors may contribute to the design of new pharmaceuticals and supply information which will facilitate the understanding of the interaction between inhibitors and the enzyme. The dried root of Salvia miltiorhiza is called 'Danshen' in China, and has been used for the treatment of cerebrovascular disease and CNS deterioration in old age for over one thousand years. In this work, a modified Ellman method was used to guide the fractionation of the active AChE inhibitory compounds from an acetone extract. Four inhibitory compounds, dihydrotanshinone, cryptotanshinone, tanshinone I and tanshinone IIA were isolated, and the structures were identified by comparison of their spectral characteristics with previous reports. The inhibitory activities of dihydrotanshinone and cryptotanshinone were dose dependent, their IC (50) values being 1.0 microM and 7.0 microM, respectively. These two compounds were the major inhibitory compounds in the extract as judged by HPLC analysis, forming 0.054 % w/w and 0.23 % w/w in the dried root, respectively, and in mixture they appear to be less active than as isolated compounds. The clogP values of dihydrotanshinone, cryptotanshinone, tanshinone I and tanshinone IIA were calculated as 2.4, 3.4, 4.8 and 5.8, respectively, which indicate that these compounds have potential to penetrate the blood-brain barrier. This is the first example of diterpenoids as inhibitors of AChE. PMID- 15114494 TI - New Polyacetylenes, DGAT inhibitors from the roots of Panax ginseng. AB - The petroleum ether extract of Panax ginseng showed a significant inhibition of the diacylglycerol acyltransferase (DGAT) enzyme from rat liver microsomes. Bioactivity-guided fractionation led to the isolation of two new polyacetylenic compounds, (9 R,10 S)-epoxyheptadecan-4,6-diyn-3-one ( 1) and 1-methoxy-(9 R,10 S)-epoxyheptadecan-4,6-diyn-3-one ( 2). Their chemical structures were elucidated on the basis of spectroscopic evidence and asymmetric synthesis. IC50 values of 9 microg/mL ( 1) and 32 microg/mL ( 2) were obtained. PMID- 15114496 TI - In vitro effects of two extracts and two pure alkaloid preparations of Uncaria tomentosa on peripheral blood mononuclear cells. AB - In the traditional Peruvian medicine, hot aqueous extracts of Uncaria tomentosa have been used for the treatment of a wide range of health problems, particularly digestive complaints and arthritis. Some of the beneficial effects observed in patients suggest an immunomodulatory capacity of Uncaria tomentosa extracts. In this study, the effects of two extracts and two mixtures of tetracyclic and pentacyclic oxindole alkaloids of Uncaria tomentosa were investigated in freshly isolated human peripheral blood mononuclear cells (PBMC) stimulated with the mitogens phytohaemagglutinin (PHA) and concanavalin A (Con A) in vitro. Neopterin production and tryptophan degradation were monitored in culture supernatants to determine the effects of the test substances on immunobiochemical pathways induced by interferon-gamma. Compared to unstimulated cells PHA and Con A increased the production of neopterin and degradation of tryptophan (p < 0.01). HCl and ethanol extracts and mixtures of alkaloids of Uncaria tomentosa inhibited both effects in a dose-dependent manner, the lowest effective concentrations of the extracts were 500 - 1000 microg/mL and of the alkaloid mixtures 100 - 175 microg/mL (p < 0.05 and < 0.01). With the highest concentrations of extracts and mixtures complete suppression of mitogen-induced neopterin production and tryptophan degradation was observed. These data demonstrate that Uncaria tomentosa extracts and mixtures of alkaloids modulate the immunobiochemical pathways induced by interferon-gamma. The findings imply a potential application of the extracts as immunoregulators and would be in line with observations in patients using these extracts. PMID- 15114498 TI - In vivo antiosteoporotic activity of a fraction of Dioscorea spongiosa and its constituent, 22-O-methylprotodioscin. AB - The antiosteoporotic activity of the 90 % EtOH fraction of the water extract of rhizomes of Dioscorea spongiosa and methylprotodioscin, its major constituent, were examined in the model of postmenopausal bone loss using ovariectomized (OVX) rats or mice. After 6 weeks treatment, the proximal tibia of rats or mice and the distal femora of mice were scanned by peripheral quantitative computed tomography (pQCT). Both the 90 % EtOH fraction (100 mg/kg/d) and methylprotodioscin (50 mg/kg/d) significantly inhibited bone loss in bone mineral content (BMC) and bone mineral density (BMD) in total, cancellous and cortical bones, and the decrease in bone strength indexes induced by OVX, without side effect on the uterus. PMID- 15114497 TI - Antitumor and antimetastatic activities of 4-hydroxyderricin isolated from Angelica keiskei roots. AB - The roots of Angelica keiskei Koizumi (Umbelliferae) have traditionally been used as a health food considered to have diuretic, laxative, analeptic and lactagogue effects. Recently, it has been thought that the roots and herbs of A. keiskei have preventive effects against coronary heart disease, hypertension and cancer. It has been reported that chalcone derivatives, such as xanthoangelol and 4 hydroxyderricin, are isolated as main components from this root. Recently, we reported that the 50 % ethanol extract, the ethyl acetate-soluble fraction and the isolated xanthoangelol, inhibited tumor growth and metastasis to the lung in Lewis lung carcinoma (LLC)-bearing mice. In the present study, we examined the effects of 4-hydroxyderricin on tumor growth and metastasis to the lung or liver in subcutaneous or intrasplenic LLC-implanted C57BL/6J female mice. 4 Hydroxyderricin at a dose of 50 mg/kg x 2/day orally inhibited the tumor growth in subcutaneous LLC-implanted mice and inhibited the lung metastasis and prolonged the survival time in mice after the removal of subcutaneous tumors by surgical operation. Doxorubicin (5 mg/kg x 2/week, i. p.) inhibited the tumor growth and metastasis to the lung, but it shortened the survival time and reduced the survival rate compared to those in 4-hydroxyderricin-treated mice. 4 Hydroxyderricin inhibited DNA synthesis in LLC cells at a concentration of 100 microM, but it had no effect on the DNA synthesis in human umbilical vein endothelial cells (HUVECs) or on the adherence of LLC cells to HUVECs. 4 Hydroxyderricin inhibited Matrigel-induced formation of capillary-like tubes by HUVECs at concentrations of 10 to 100 microM. The weights of the spleen and thymus in mice with subcutaneously implanted LLC were maintained close to those of normal mice by orally administered 4-hydroxyderricin. In addition, 4 hydroxyderricin (50 mg/kg x 2/day) inhibited the reduction of the numbers of lymphocytes, CD4+, CD8+ and natural killer (NK)-T cells in the spleen of tumor removed mice. Doxorubicin reduced the numbers of lymphocytes, CD4+, CD8+ and NK cells compared to those in LLC-removed mice. These results suggest that the antitumor and antimetastatic activities of 4-hydroxyderricin may be modulated by the immune system and the inhibition of angiogenesis. PMID- 15114499 TI - In vitro behaviour of sesquiterpene lactones and sesquiterpene lactone-containing plant preparations in human blood, plasma and human serum albumin solutions. AB - The interactions of the three sesquiterpene lactones (SLs) dihydrohelenalin acetate, dihydrohelenalin methacrylate and helenalin isobutyrate from Arnica montana and of parthenolide from Tanacetum parthenium as well as of three ethanolic Arnica preparations with human blood proteins using different matrices, human serum albumin (HSA), plasma and whole blood, were investigated. The extent of protein binding in human plasma or to human albumin differed significantly between individual SLs (dihydrohelenalin methacrylate < dihydrohelenalin acetate < helenalin isobutyrate << parthenolide), e. g., 30 % to 50 % of the SLs were bound to plasma. On the whole, SLs in the ethanolic preparations showed a lower degree of protein binding. Interestingly, although HSA has a reactive thiol group at its cysteine-34 position which is prone to react with the alpha,beta unsaturated carbonyl of SLs, our studies showed that less than 6 % of SLs are bound to this position within 60 minutes. Thus, a reaction with other amino acids as well as non-covalent interactions with plasma proteins have to be considered. Considering the biological activity of SLs in whole blood, our studies demonstrate that knowledge of their plasma protein binding is important for interpreting the reported data of in vitro and ex vivo assays. PMID- 15114500 TI - New lignans and cytotoxic constituents from Wikstroemia lanceolata. AB - Two new lignans, (-)-aptosimon ( 1) and (-)-diasesamin-di-gamma-lactone ( 2), together with twenty-two known compounds, have been isolated from the stems and roots of Wikstroemia lanceolata. The structures of the new compounds were determined through spectral analyses. The proton and carbon assignments of the known sesquiterpenoid, 1alpha,7alpha,10alpha H-guaia-4,11-dien-3-one ( 24) were revised by 2D NMR techniques. Among the isolates, three compounds, (+)-hinokinin ( 6), 2,6-dimethoxy- p-benzoquinone ( 17), and 1alpha,7alpha,10alpha H-guaia-4,11 dien-3-one ( 24) exhibited effective cytotoxicities (ED50 values < 4.0 microg mL 1) against P-388 and HT-29 tumor cell lines. PMID- 15114501 TI - New eremophilane-type sesquiterpenes from Ligularia lapathifolia. AB - Seven new eremophilanolides were isolated from the roots and rhizomes of Ligularia lapathifolia. Their structures were established as 3beta-angeloyloxy 8beta H-eremophil-7(11)-ene-12,8alpha (14beta, 6alpha)-diolide, 3beta-angeloyloxy 8beta-hydroxyeremophil-7(11)-ene-12,8alpha(14beta,6alpha)-diolide,3beta angeloyloxy-8beta-methoxyeremophil- 7(11)-ene-12,8alpha(14beta,6alpha) diolide,3beta-angeloyloxy-8beta-ethoxyeremophil-7(11)-ene-12, 8alpha (14beta,6alpha)-diolide, 3beta-angeloyloxy-10beta- hydroxyeremophil-8(9),7(11) diene-12,8(14beta,6alpha)-diolide, 3beta-angeloyloxy-8,12-expoy-12alpha-hydroxy 8beta-methoxyeremophil-7(11)-en-14beta,6alpha-olide and 3beta angeloyloxyeremophilan-7,11-dien-14beta,6alpha-olide, by means of spectroscopic analyses. Moreover, application of a photooxygenation reaction on 7 resulted in the generation of 2 with an alpha,beta-unsaturated gamma-lactone moiety. This biomimetic transformation supports a biogenetic pathway proposed for 2. PMID- 15114502 TI - Euphopubescenol and euphopubescene, two new jatrophane polyesters, and lathyrane type diterpenes from Euphorbia pubescens. AB - The structures of euphopubescenol and euphopubescene, two new macrocyclic jatrophane diterpene polyesters, isolated from the whole dried plant of Euphorbia pubescens, were established as 5alpha,8alpha,15beta-triacetoxy-3alpha-benzoyloxy 4alpha-hydroxy -9,14-dioxo-13beta H-jatropha-6(17),11 E-diene ( 1) and 3beta,7beta,8beta,9alpha,14alpha,15beta-hexaacetoxy-2beta H-jatropha-5 E,11 E diene ( 2) by 1D- and 2D-NMR (COSY, HMQC, HMBC and NOESY), IR, EI-MS and EI-FTICR MS. Two known lathyrane derivatives, jolkinol A ( 3) and jolkinol A ( 4), whose (13)C-NMR spectra were assigned, were also isolated. Compounds 1 - 3 have been evaluated for their ability to inhibit the in vitro growth of three human tumour cell lines representing different tumour types, MCF-7 (breast adenocarcinoma), NCI-H460 (non-small cell lung cancer) and SF-268 (CNS cancer). They inhibited both MCF-7 and NCI-H460 cell lines, with GI50 values ranging between 40.9 microM and 95.3 microM, but were found to be ineffective as growth inhibitors of the SF 268 cell line. PMID- 15114503 TI - Metabolomic strategy for the classification and quality control of phytomedicine: a case study of chamomile flower (Matricaria recutita L.). AB - In order to improve the accuracy and consistency of control phytomedicine preparations worldwide, regulatory authorities are requesting research into new analytical methods for the stricter standardisation of phytomedicines. Such methods have to be both objective and robust, and should address the reproducibility of the content of the chemical profiles. NMR-based metabolomics, which combines high-resolution (1)H-NMR spectroscopy with chemometric analysis, has been employed as an innovative way to meet those demands. In this paper, chamomile flowers from three different geographical regions, namely, Egypt, Hungary and Slovakia were characterised using 1H-NMR spectroscopy followed by principal component analysis. It was found that the origin, purity and preparation methods contributed to the differences observed in prepared chamomile extracts. In addition, this method also enabled the elucidation of the molecular information embedded in the spectra responsible for the observed variability. The metabolomic strategy employed in the current study should provide an efficient tool for the quality control and authentication of phytomedicines. PMID- 15114504 TI - Inhibitory effects of ent-kauranes from the stems of Annona squamosa on superoxide anion generation by human neutrophils. AB - Eleven ent-kauranes, isolated from the fresh stems of Annona squamosa L. (Annonaceae), were subjected to assays on the generation of superoxide anion (O2. -) by human neutrophils. Except for ent-kaur-16-en-19-oic acid, 16beta,17 dihydroxy- ent-kauran-19-al, and 16alpha,17-dihydroxy -ent-kauran-19-al, all ent kauranes showed significant inhibitory effect on O2.-- generation in response to formyl- L-methionyl- L-leucyl- L-phenylalanine (fMLP/CB). In contrast, phorbol myristate acetate (PMA)-induced O2.-- generation was not suppressed by any ent kauranes. Especially, ent-kaur-16-en-19-oic acid could significantly increase O2. - production. The structure-activity relationship of these compounds is also discussed herein. Furthermore, the effect of ent-kauranes on nitric oxide generation by NR8383 macrophages in response to lipopolysaccharide (LPS) was examined. None of the compounds showed an inhibitory effect on nitric oxide generation. PMID- 15114505 TI - In vitro inhibition of diacylglycerol acyltransferase by prenylflavonoids from Sophora flavescens. AB - Four prenylflavonoids, kurarinone ( 1), a chalcone of 1, kuraridin ( 2), kurarinol ( 3), kushenol H ( 4) and kushenol K ( 5) isolated from the roots of Sophora flavescens were investigated for their inhibitory effects on diacylglycerol acyltransferase (DGAT). The flavonoids inhibited DGAT activity in a dose-dependent manner with IC50 values of 10.9 microM ( 1), 9.8 microM ( 2), 8.6 microM ( 3), 142.0 microM ( 4) and 250 microM ( 5). The prenylflavonoids without C3-OH ( 1, 2, 3) showed stronger inhibition than those with C3-OH ( 4, 5). On the other hand, flavonoids without side chains (hesperetin, naringenin, quercetin and kaempferol) did not inhibit the enzyme activity at a final concentration of 800 microM. These data suggest that the lavandulyl side chain and the position of the hydroxy group are important for high DGAT inhibitory activity. Compound 1 also inhibited de novo synthesis of triacylglycerol (TG) in Raji cells. PMID- 15114506 TI - Acetylcholinesterase enzyme inhibitory effects of amaryllidaceae alkaloids. AB - Twenty-three Amaryllidaceae alkaloids having several different ring types were evaluated for their acetylcholinesterase enzyme (AChE) inhibitory activity. The alkaloid 1- O-acetyllycorine (IC50 : 0.96 +/- 0.04) showed significant AChE inhibitory activity. In addition, crinine (IC50 : 461 +/- 14), crinamidine (IC50 : 300 +/- 27), epivittatine (IC50 : 239 +/- 9), 6-hydroxycrinamine (IC50 : 490 +/ 7), N-desmethyl-8alpha-ethoxypretazettine (IC50 : 234 +/- 13) N-desmethyl-8beta ethoxypretazettine (IC50 : 419 +/- 8), lycorine (IC50 : 213 +/- 1), and 1,2-di- O acetyllycorine (IC50 : 211 +/- 10) had weak activity. Lycorine-type alkaloids were the most active alkaloids with 1- O-acetyllycorine exhibiting inhibitory effects two-fold more potent than that of galanthamine. PMID- 15114507 TI - Lipoxygenase inhibition by anadanthoflavone, a new flavonoid from the aerial parts of Anadenanthera colubrina. AB - Chemical investigation of the aerial parts of Anadenanthera colubrina led to the isolation of a new flavonoid named anadanthoflavone ( 1), along with 11 known compounds: alnusenol, lupenone, lupeol, betulinic acid, alpha-amyrin, beta amyrin, beta-sitosterol, stigmasterol, apigenin, 4-hydroxybenzoic acid and cinnamic acid. The isolated compounds were evaluated for their inhibitory activity on human platelet 12-lipoxygenase (12-hLO), human reticulocyte 15 lipoxygenase (15-hLO) and soybean lipoxygenase-1 (15-sLO). Compound 1 was found to be active against 12-hLO and 15-hLO with IC50 values of 13 +/- 3 microM and 17 +/- 3 microM, respectively. Apigenin selectively inhibited the activity of 15-hLO (IC50 : 4.0 +/- 1 microM), while lupenone, lupeol and alpha-amyrin were found active against 15-sLO (IC50 : 22 +/- 3 microM, 35 +/- 9 microM and 15 +/- 3 microM, respectively). PMID- 15114508 TI - Bisbenzyltetrahydroisoquinolines, a new class of inhibitors of the mitochondrial respiratory chain complex I. AB - Four bisbenzyltetrahydroisoquinoline alkaloids (-)-medelline, (+)-antioquine, (+) aromoline, and (+)-obamegine were isolated from the fruits of Xylopia columbiana. These compounds, the previously isolated alkaloids (+)-thaligrisine and (+) isotetrandrine, as well as their O-acetylated derivatives were assayed on submitochondrial particles from beef heart as inhibitors of the mammalian respiratory chain. The results revealed that these alkaloids act as selective inhibitors of mitochondrial complex I in a 0.15 - 4.71 microM range. O Acetylation, which increases their lipophilicity, considerably increased the inhibitory potency. PMID- 15114509 TI - Cytotoxic ent-kaurane diterpenoids from Isodon rubescens var. rubescens. AB - Five new ent-kauranoids, xindongnins C - G ( 1 - 5), together with five known ones, xindongnins A and B ( 6 and 7), melissoidesin G ( 8), dawoensin A ( 9), and glabcensin V ( 10), were isolated from the leaves of Isodon rubescens var. rubescens, and structurally elucidated. Compounds 1, 4, and 6 - 10 showed inhibitory effects against human tumor K562 cells with IC (50) values ranging from 0.3 to 7.3 microg/mL. The structure of 6 was revised on the basis of its 2D NMR spectral data. PMID- 15114510 TI - Antimicrobial activity of 3-O-acyl-(-)-epicatechin and 3-O-acyl-(+)-catechin derivatives. AB - As an exploratory investigation of antimicrobial promoting compounds, 3- O-acyl-( )-epicatechins and 3- O-acyl-(+)-catechins possessing various aromatic groups and aliphatic chains of varying length from C4 to C16 for increasing lipophilicity were synthesized and tested for antimicrobial activities against Gram-positive, Gram-negative bacteria and fungi. The (-)-epicatechin and (+)-catechin derivatives comprised of aromatic groups increased activity and derivatives with acyl chain groups of carbon atoms in the close vicinity of C8 to C10 showed strong antimicrobial activity (MIC = 2 - 8 microg/ml) against Gram-positive bacteria and weak activity against fungi. However, the activity decreased when the carbon chain length of the substituents was too short (C4 to C6) or too long (C16). These results suggest that the presence of lipophilic substituents with moderate sizes might be crucial for the optimal antimicrobial activity. PMID- 15114511 TI - Antitubercular and antiplasmodial constituents of Abrus precatorius. AB - A known isoflavanquinone, abruquinone B ( 1), and a new derivative, abruquinone G ( 2), were isolated from the aerial parts of Abrus precatorius. Chemical structures of these compounds were elucidated by spectral analyses. While 1 exhibited antitubercular, antiplasmodial and cytotoxic activities, compound 2 showed mild antiviral and cytotoxic activities. PMID- 15114512 TI - p-Anisaldehyde: acaricidal component of Pimpinella anisum seed oil against the house dust mites Dermatophagoides farinae and Dermatophagoides pteronyssinus. AB - The acaricidal activity of anise seed oil-derived p-anisaldehyde and commercially available components of anise seed oil was examined against Dermatophagoides farinae and D. pteronyssinus and compared with those of the synthetic acaricides, benzyl benzoate and N,N-diethyl- m-toluamide (DEET). On the basis of LD 50 values, the compound most toxic to D. farinae adults was p-anisaldehyde (1.11 microg/cm2) followed by benzyl benzoate (9.32 microg/cm2), DEET (36.84 microg/cm2), 3-carene (42.10 microg/cm2), and estragol (43.23 microg/cm2). Against D. pteronyssinus adults, p-anisaldehyde (0.98 microg/cm2) was much more effective than benzyl benzoate (6.54 microg/cm2), DEET (17.79 microg/cm2), 3 carene (39.84 microg/cm 2), and estragol (40.11 microg/cm2). p-Anisaldehyde was about 8.4 and 6.7 times more toxic than benzyl benzoate against D. farinae and D. pteronyssinus adults, respectively. The results suggested that p-anisaldehyde may be useful as a lead compound for the development of new agents for the selective control of house dust mites. PMID- 15114513 TI - Kaurane-type diterpene glycoside from the stem bark of Acanthopanax trifoliatus. AB - A new diterpene glycoside, acantrifoside D ( 1), as well as three known diterpenes (2 - 4) were isolated from the stem bark of Acanthopanax trifoliatus from Vietnam. Based on 1D and 2D NMR spectroscopic data, their chemical structures were determined to be 16alpha,17-dihydroxy- ent-kauran-19-oic acid 16- O-beta- D-glucopyranoside 19- O-beta- D-glucopyranosyl ester (1), 16alpha H,17 isovalerate- ent-kauran-19-oic acid (2), ent-kaur-16-en-19-oic acid ( 3), and ent pimara-8(14),15-dien-19-oic acid (4). Compounds 2 - 4 had strong inhibitory effects against COX-1 in an in vitro assay, with IC50 values of 0.21, 0.15 and 0.19 mM, respectively. PMID- 15114514 TI - Determination of six major flavonoid glycosides in Saussurea mongolica by capillary electrophoresis. AB - A new capillary electrophoresis (CE) method was established for simultaneous assay of six flavonoid O-glycosides, asebotin (AS), kaempferol 3- O-beta- D glucopyranoside (KG), kaempferol 3- O-alpha- L-rhamnopyranoside (KR), 7 methoxykaempferol 3- O-alpha- L-rhamnopyranoside (KMR), quercetin 3- O-beta- D glucopyranoside (QG) and quercetin 3- O-alpha- L-rhamnopyranoside (QR) obtained from Chinese herbal plant extract of Saussurea mongolica. The optimum buffer system was 30 mmol/L borate buffer (Na2B4O7/HCl, pH 9.00) with 40 % (v/v) methanol. Voltage was 15 kV and detection at 270 nm. The application of this method for the separation and determination of the six flavonoid O-glycosides in S. mongolica is reported. The relative standard deviations of migration times and peak areas were < 2.61 and 4.41%, respectively. The contents of six natural products ranged from 0.043 to 0.60 mg/g and recoveries ranged from 92.4 to 104.4%. The effects of pH value, buffer concentration, surfactant, beta cyclodextrin and organic modifier on the separation were investigated. PMID- 15114515 TI - Characterization of medicinal Epimedium species by 5S rRNA gene spacer sequencing. AB - Sequences of 5S rRNA gene spacer were used to identify Epimedium brevicornu Maxim., E. sagittatum (Sieb. et Zucc.) Maxim., E. wushanense T. S. Ying, E. pubescens Maxim., and E. koreanum Nakai. These species are listed as source plants of Chinese medicine 'Ying Yang Huo' in the Chinese Pharmacopoeia. The neighbor-joining method was used in a sequence analysis of Epimedium species. A position-specific nucleotide was found in the 5S rRNA gene spacer for E. pubescens, E. wushanense, and E. brevicornu. A 19-bp deletion was found for E. koreanum in the 5S rRNA gene spacer. E. koreanum was most divergent from the other four endemic Chinese species of Epimedium. PMID- 15114516 TI - Testosterone regulates mRNA levels of calcium regulatory proteins in cardiac myocytes. AB - Gender-related differences in cardiac function have been described in the literature, but whether the presence of sex hormones is responsible for these differences remains unclear. This study was designed to determine whether testosterone regulates the gene expression of calcium regulatory proteins in rat heart, thus playing a role in gender-related differences in cardiac performance. Ventricular myocytes were isolated from two-day-old rats and treated with testosterone at varying duration; the levels of gene expression for the androgen receptor (AR) and major calcium regulatory proteins were determined by quantitative real-time PCR. Testosterone (1 microM) treatment induced a maximum increase in beta1-adrenergic receptor and L-type calcium channel mRNA levels following an eight hour exposure. Six hours testosterone treatment stimulated a 300-fold increase in androgen receptor message abundance, and Na/Ca exchanger mRNA levels reached a maximum level following twenty-four hour testosterone treatment. Taken together, these data provide the first evidence that testosterone regulates gene expression of the major calcium regulatory proteins in isolated ventricular myocytes, and may thus play a role in the gender-related differences observed in cardiac performance. PMID- 15114518 TI - Signals in the activation of opioid mu-receptors by loperamide to enhance glucose uptake into cultured C2C12 cells. AB - In an attempt to understand the signal pathways of opioid mu-receptors for glucose metabolism, we used loperamide to investigate the glucose uptake into the myoblast C2C12 cells. Loperamide enhanced the uptake of radioactive deoxyglucose into C2C12 cells in a concentration-dependent manner that was abolished in cells pre-incubated with naloxone or naloxonazine at concentrations sufficient to block opioid mu-receptors. Pharmacological inhibition of phospholipase C (PLC) by U73122 resulted in a concentration-dependent decrease in loperamide-stimulated uptake of radioactive deoxyglucose into C2C12 cells. This inhibition of glucose uptake by U73122 was specific since the inactive congener, U73343, failed to modify loperamide-stimulated glucose uptake. Moreover, both chelerythrine and GF 109203X diminished the action of loperamide at concentrations sufficient to inhibit protein kinase C (PKC). The obtained data suggest that an activation of opioid mu-receptors in C2C12 cells by loperamide may increase glucose uptake via the PLC-PKC pathway. PMID- 15114517 TI - No in vitro effects of fatty acids on glucose uptake, lipolysis or insulin signaling in rat adipocytes. AB - Elevated plasma levels of free fatty acids (FFA) can produce insulin resistance in skeletal muscle tissue and liver and, together with alterations in beta-cell function, this has been referred to as lipotoxicity. This study explores the effects of FFAs on insulin action in rat adipocytes. Cells were incubated 4 or 24 h with or without an unsaturated FFA, oleate or a saturated FFA, palmitate (0.6 and 1.5 mM, respectively). After the culture period, cells were washed and insulin effects on glucose uptake and lipolysis as well as cellular content of insulin signaling proteins (IRS-1, PI3-kinase, PKB and phosphorylated PKB) and the insulin regulated glucose transporter GLUT4 were measured. No significant differences were found in basal or insulin-stimulated glucose uptake in FFA treated cells compared to control cells, regardless of fatty acid concentration or incubation period. Moreover, there were no significant alterations in the expression of IRS-1, PI3-kinase, PKB and GLUT4 following FFA exposure. Insulin's ability to stimulate PKB phosphorylation was also left intact. Nor did we find any alterations following FFA exposure in basal or cAMP-stimulated lipolysis or in the ability of insulin to inhibit lipolysis. The results indicate that oleate or palmitate does not directly influence insulin action to stimulate glucose uptake and inhibit lipolysis in rat fat cells. Thus, lipotoxicity does not seem to occur in the fat tissue itself. PMID- 15114519 TI - Lipid and lipoprotein profile in menopausal transition. Effects of hormones, age and fat distribution. AB - The behavior of lipoproteins during the menopausal transition and their relationship with sex hormones and body fat distribution is still unclear. Our aim was to evaluate atherogenic IDL, LDL, Lp(a) and antiatherogenic HDL lipoproteins in four groups of women: premenopausal (n = 20), menopausal transition women with menstrual bleeding (n = 31), menopausal transition women with 3 to 6 months amenorrhea (n = 36), and postmenopausal women (n = 30). We also measured their FSH, LH and estradiol levels along with BMI and waist circumference. Menopausal transition and postmenopausal women showed higher values of waist circumference (p < 0.0032), LDL-cholesterol (p < 0.002), IDL cholesterol (p < 0.002) and apoprotein B (p < 0.0001) than premenopausal women. Total-cholesterol (p < 0.0001), triglycerides (p < 0.004), IDL-cholesterol and Lp(a) were higher in menopausal transition women with amenorrhea and in postmenopausal women in comparison with premenopausal women. After adjustment according to age and waist circumference, multiple regression analysis showed the increase in total-cholesterol and LDL-cholesterol to be linearly associated to menopausal status and estradiol concentration, whereas Lp(a) was only related to menopausal status. Age was found to be an independent variable in relation to apoprotein B concentration changes. The effect of menopausal status on TG levels did not remain in the model when age, waist and BMI were included (beta = 0.05, p = 0.356). HDL-cholesterol levels were the same in all the groups. Menopause, age and the increase in abdominal fat distribution were three independent and significant factors impairing lipoprotein profiles from the beginning of the menopausal transition. PMID- 15114520 TI - Effects of carnitines on rat sertoli cell protein metabolism. AB - The influence of carnitine on male reproductive function is well established. However, the molecular mechanism by which carnitines control male fertility have not yet been clearly identified. In the present study, we studied the role of carnitines at testis level by examining their effects on protein metabolism in cultured rat Sertoli cells. Our results demonstrate that L-carnitine and, in particular, L-acetylcarnitine administration affects overall protein synthesis, inducing a dose-dependent decrease in amino acid incorporation and consequently an increase of the intracellular amino-acid pool useful to speed up Sertoli cell energy metabolism. We also observed significant effect on the expression of specific proteins involved in the regulation of glucose uptake and Insulin-like Growth Factor (IGF) biological activity. Actually, an increased expression of the mRNA coding for the membrane Glucose Transporter-1 (GLUT-1) and a decreased expression of mRNA coding for the negative modulator insulin-like growth factor binding protein-4 (IGFBP-4) were induced by in vitro treatment with L-carnitine and L- acetylcarnitine separately and in combination; in fact, the effects observed were far more pronounced following concomitant treatment with both compounds. Taken together, these data indicate that the influence of carnitines on male fertility could be partly mediated by an activity at Sertoli cell level. PMID- 15114521 TI - Effects of PPARgamma and PPARalpha agonists on serum leptin levels in diet induced obese rats. AB - Leptin and peroxisome proliferator-activated receptors are two important adipose tissue factors involved in energy metabolism regulation. It has been shown that PPARgamma agonists decrease leptin levels. However, the effects of PPARalpha agonists on leptin have not been investigated much. The aim of this study was to compare the effects of a PPARgamma agonist rosiglitazone (RSG) and PPARalpha agonist gemfibrozil (G) on body weight and serum insulin and leptin levels in diet-induced obese rats. Male Wistar rats were divided into six groups according to diet and drug therapy. After four weeks, serum glucose, triglyceride, insulin and leptin levels were significantly decreased in the high-fat-fed and RSG treated groups compared to the group fed a high-fat diet only (162 +/- 19 vs. 207 +/- 34 mg/dl, 58 +/- 20 vs. 112 +/- 23 mg/dl, 3.1 +/- 1.0 vs. 15.2 +/- 4.0 ng/ml, 1.6 +/- 0.5 vs. 3.6 +/- 1.6 ng/ml, respectively). However, these parameters were not statistically different in RSG animals treated with a standard diet compared to the standard diet group. The high fat+RSG group gained much more weight compared to high-fat and high-fat+G groups (p > 0.05). Additionally, serum glucose, insulin and leptin levels were significantly decreased in the high-fat fed and G-treated group compared to high-fat group (149 +/- 19 vs. 207 +/- 34 mg/dl, 57 +/- 16 vs. 112 +/- 23 mg/dl, 4.3 +/- 2.1 vs. 15.2 +/- 4.0 ng/ml, 1.6 +/ 0.4 vs. 3.6 +/- 1.6 ng/ml, respectively). These results suggest that PPARalpha agonists may decrease serum glucose, insulin and leptin levels as PPARgamma agonists do in diet-induced obese rats. PMID- 15114522 TI - Effect of purine-free low-malt liquor (happo-shu) on the plasma concentrations and urinary excretion of purine bases and uridine--comparison between purine-free and regular happo-shu. AB - To determine whether purine-free and regular low-malt liquor beverages (happo shu) increase the plasma concentration and urinary excretion of purine bases (hypoxanthine, xanthine, uric acid) and uridine, 6 healthy males were given regular (10 ml/kg of body weight) and purine-free happo-shu (10 ml/kg of body weight). Plasma concentration-time curves were plotted, and the areas under the curves for uric acid and total purine bases (the sum of hypoxanthine, xanthine, and uric acid) were greater in the regular than in the purine-free happo-shu ingestion experiment (both p < 0.05). In addition, the total urinary excretion of xanthine, total purine bases, and uridine was greater in the regular than in the purine-free happo-shu ingestion experiment (p < 0.05 in all cases), although the total urinary excretion of hypoxanthine and uric acid was no different between the regular and the purine-free happo-shu ingestion experiments. These results suggest that uridine contained in regular happo-shu might contribute to an increase in the urinary excretion of uridine along with ethanol, and that the purines contained in regular happo-shu may contribute to the increase in plasma concentration of uric acid due to purine degradation. PMID- 15114523 TI - Relationship between adiponectin and metabolic variables in Caribbean offspring of patients with type 2 diabetes mellitus. AB - AIM: To examine the relationship between adiponectin and metabolic variables in the offspring of patients with type 2 diabetes mellitus. METHODS: Fasting blood samples and anthropometric indices were taken from 34 subjects, offspring of patients with type 2 diabetes, and 24 healthy control subjects without any immediate family history of diabetes. Plasma glucose and serum adiponectin, insulin, triglycerides, total cholesterol, HDL and LDL cholesterol levels were measured, and insulin resistance (IR) was calculated based on the homeostasis model assessment (HOMA) method. RESULTS: Offspring and control subjects were sex matched, but the offspring were older and had higher body mass index and waist circumference than the control subjects (p < 0.05). The offspring had significantly higher mean fasting plasma glucose concentrations; however, their mean serum insulin, adiponectin, triglyceride, total cholesterol, HDL and LDL cholesterol and HOMA-derived IR levels did not significantly differ from those of the control subjects (p > 0.05). While the negative correlation between serum adiponectin and HDL cholesterol levels in the offspring remained statistically significant after adjusting for the effect of age, sex and BMI (r = -0.37, p < 0.05), the negative correlation between adiponectin and serum triglyceride, LDL cholesterol or IR levels became non-significant after controlling for the above variables (p > 0.05 in all cases). CONCLUSION: The correlation between adiponectin and some known biochemical risk factors for developing diabetes and cardiovascular disease in the offspring of patients with diabetes warrants further study to evaluate its potential in assessing the risk of developing these disorders. PMID- 15114524 TI - Bradykinin B2 receptor gene C-58T polymorphism and insulin resistance. A study on obese patients. AB - The bradykinin B2 receptor (B2R) gene is a candidate in the pathogenesis of insulin resistance, which often clusters with other abnormalities in metabolic syndrome. We investigated the distribution of the C-58T B2R gene polymorphism within a population of overweight/obese patients (BMI > or = 25 kg/m2) potentially characterised by different levels of insulin resistance. Patients with type 2 diabetes, dyslipidemia and hypertension were excluded in order to distinguish the effect of obesity on insulin sensitivity from that of confounding factors. Ninety-two unrelated adults (41 men and 51 women, aged 33.7 +/- 11.6 years) were recruited by random sampling from a general population evaluated for cardiovascular risk stratification. Measurements included BMI, waist circumference, body composition, blood pressure, serum leptin, and lipid profile. Insulin sensitivity was calculated according to the homeostasis model assessment (HOMA) method. C-58T genotypes--CC (n = 20), CT (n = 47) and TT (n = 25)--were determined by restriction fragment-length polymorphism PCR. Patients subdivided on the basis of C-58T polymorphism, showed no difference in any of the parameters examined, including HOMA index values, after adjustment for age, sex, BMI and waist circumference. The results indicate that the C-58T B2R gene polymorphism is not associated with different levels of insulin resistance within a population of obese patients. PMID- 15114525 TI - The effect of gender and age on growth hormone replacement in growth hormone deficient patients. AB - We analyzed the effect of growth hormone replacement therapy (36 months) analyzed at a dose adjusted to maintain serum insulin-like growth factor-I level between the median and the upper end of the age-related reference range on bone mineral density, body composition, and carbohydrate metabolism with respect to gender and age in 20 adult patients (9 women, 11 men, mean age: 43 years, range: 21-61 years). The lumbar and femoral T-score was increased after 12 and after 18 months of therapy respectively in men (p < 0.001 and p = 0.002), but did not changed significantly in women. The increase of femoral T-score was greater in young men (< or = 45 years, n = 6) than old men (> 45 years, n = 5, p < 0.001). Body fat was lower in men than in women after 6 months (p = 0.002). The waist/hip ratio only decreased in women (p = 0.044). The waist circumference decreased in both genders after 6 months of therapy (p < 0.001), but more markedly in females than in males (p < 0.05). The sum of skinfold thicknesses was reduced in males after 6 months of therapy (p < 0.001). Changes in body composition parameters measured were independent of age. The glycosylated hemoglobin increased without sex or age difference after 12 months of initiation of therapy (p < 0.001), but fasting glucose and insulin levels did not change during the therapy. Our results indicate that the effect of growth hormone replacement on bone mineral content in adults is age- and gender-dependent, gender dependent on body composition, but independent of age and gender on carbohydrate metabolism. PMID- 15114526 TI - Elevation of breast milk leptin levels by laughter. PMID- 15114527 TI - Nomegestrol acetate: a progestin that deserves recognition. PMID- 15114529 TI - Insulin receptor splicing alteration in myotonic dystrophy type 2. AB - Myotonic dystrophy (DM) is caused by either an untranslated CTG expansion in the 3' untranslated region of the DMPK gene on chromosome 19 (dystrophia myotonica type 1 [DM1]), or an untranslated CCTG tetranucleotide repeat expansion in intron 1 of the ZNF9 gene on chromosome 3 (dystrophia myotonica type 2 [DM2]). RNA binding proteins adhere to transcripts of the repeat expansions that accumulate in the nucleus, and a trans-dominant dysregulation of pre-mRNA alternative splicing has been demonstrated for several genes. In muscle from patients with DM1, altered insulin-receptor splicing to the nonmuscle isoform corresponds to the insulin insensitivity and diabetes that are part of the DM phenotype; because of insulin-receptor species differences, this effect is not seen in mouse models of the disease. We now demonstrate that comparable splicing abnormalities occur in DM2 muscle prior to the development of muscle histopathology, thus demonstrating an early pathogenic effect of RNA expansions. PMID- 15114530 TI - AICA-ribosiduria: a novel, neurologically devastating inborn error of purine biosynthesis caused by mutation of ATIC. AB - In a female infant with dysmorphic features, severe neurological defects, and congenital blindness, a positive urinary Bratton-Marshall test led to identification of a massive excretion of 5-amino-4-imidazolecarboxamide (AICA) riboside, the dephosphorylated counterpart of AICAR (also termed "ZMP"), an intermediate of de novo purine biosynthesis. ZMP and its di- and triphosphate accumulated in the patient's erythrocytes. Incubation of her fibroblasts with AICA-riboside led to accumulation of AICAR, not observed in control cells, suggesting impairment of the final steps of purine biosynthesis, catalyzed by the bifunctional enzyme AICAR transformylase/IMP cyclohydrolase (ATIC). AICAR transformylase was profoundly deficient, whereas the IMP cyclohydrolase level was 40% of normal. Sequencing of ATIC showed a K426R change in the transformylase region in one allele and a frameshift in the other. Recombinant protein carrying mutation K426R completely lacks AICAR transformylase activity. PMID- 15114531 TI - Genetic signatures of strong recent positive selection at the lactase gene. AB - In most human populations, the ability to digest lactose contained in milk usually disappears in childhood, but in European-derived populations, lactase activity frequently persists into adulthood (Scrimshaw and Murray 1988). It has been suggested (Cavalli-Sforza 1973; Hollox et al. 2001; Enattah et al. 2002; Poulter et al. 2003) that a selective advantage based on additional nutrition from dairy explains these genetically determined population differences (Simoons 1970; Kretchmer 1971; Scrimshaw and Murray 1988; Enattah et al. 2002), but formal population-genetics-based evidence of selection has not yet been provided. To assess the population-genetics evidence for selection, we typed 101 single nucleotide polymorphisms covering 3.2 Mb around the lactase gene. In northern European-derived populations, two alleles that are tightly associated with lactase persistence (Enattah et al. 2002) uniquely mark a common (~77%) haplotype that extends largely undisrupted for >1 Mb. We provide two new lines of genetic evidence that this long, common haplotype arose rapidly due to recent selection: (1) by use of the traditional F(ST) measure and a novel test based on p(excess), we demonstrate large frequency differences among populations for the persistence associated markers and for flanking markers throughout the haplotype, and (2) we show that the haplotype is unusually long, given its high frequency--a hallmark of recent selection. We estimate that strong selection occurred within the past 5,000-10,000 years, consistent with an advantage to lactase persistence in the setting of dairy farming; the signals of selection we observe are among the strongest yet seen for any gene in the genome. PMID- 15114532 TI - Extended linkage disequilibrium surrounding the hemoglobin E variant due to malarial selection. AB - The hemoglobin E variant (HbE; ( beta )26Glu-->Lys) is concentrated in parts of Southeast Asia where malaria is endemic, and HbE carrier status has been shown to confer some protection against Plasmodium falciparum malaria. To examine the effect of natural selection on the pattern of linkage disequilibrium (LD) and to infer the evolutionary history of the HbE variant, we analyzed biallelic markers surrounding the HbE variant in a Thai population. Pairwise LD analysis of HbE and 43 surrounding biallelic markers revealed LD of HbE extending beyond 100 kb, whereas no LD was observed between non-HbE variants and the same markers. The inferred haplotype network suggests a single origin of the HbE variant in the Thai population. Forward-in-time computer simulations under a variety of selection models indicate that the HbE variant arose 1,240-4,440 years ago. These results support the conjecture that the HbE mutation occurred recently, and the allele frequency has increased rapidly. Our study provides another clear demonstration that a high-resolution LD map across the human genome can detect recent variants that have been subjected to positive selection. PMID- 15114533 TI - Graphical modeling of the joint distribution of alleles at associated loci. AB - Pairwise linkage disequilibrium, haplotype blocks, and recombination hotspots provide only a partial description of the patterns of dependences and independences between the allelic states at proximal loci. On the gross scale, where recombination and spatial relationships dominate, the associations can be reasonably described in these terms. However, on the fine scale of current high density maps, the mutation process is also important and creates associations between loci that are independent of the physical ordering and that can not be summarized with pairwise measures of association. Graphical modeling provides a standard statistical framework for characterizing precisely these sorts of complex stochastic data. Although graphical models are often used in situations in which assumptions lead naturally to specific models, it is less well known that estimation of graphical models is also a developed field. We show how decomposable graphical models can be fitted to dense genetic data. The objective function is the maximized log likelihood for the model penalized by a multiple of the model's degrees of freedom. We also describe how this can be modified to incorporate prior information of locus position. Simulated annealing is used to find good solutions. Part of the appeal of this approach is that categorical phenotypes can be included in the same analysis and association with polymorphisms can be assessed jointly with the interlocus associations. We illustrate our method with genotypic data from 25 loci in the ELAC2 gene. The results contain third- and fourth-order locus interactions and show that, at this density of markers, linkage disequilibrium is not a simple function of physical distance. Graphical models provide more flexibility to express these features of the joint distribution of alleles than do monotonic functions connecting physical and genetic maps. PMID- 15114537 TI - K/DOQI clinical practice guidelines on hypertension and antihypertensive agents in chronic kidney disease. PMID- 15114547 TI - Pulmonary hemorrhage/hemoptysis in children. AB - Pulmonary hemorrhage and hemoptysis are uncommon in childhood, and the frequency with which they are encountered by the pediatric pulmonologist depends largely on the special interests of the center to which the child is referred. In those centers caring for children with cystic fibrosis or congenital heart disease, these will be by far the most common causes of hemoptysis. Other causes of hemoptysis are far less common, such as bleeding from localized lesions in the upper airway or tracheobronchial tree. Even less common is bleeding into the lungs as part of a systemic disease, usually with renal involvement (pulmonary renal syndromes), such as systemic lupus erythematosis or Goodpasture's syndrome. Bleeding into the lungs in children with a bleeding diathesis probably only occurs in immunosuppressed children after transplantation. When no other cause is found for pulmonary hemorrhage, the presumed diagnosis is idiopathic pulmonary hemosiderosis. This review discusses the various causes of hemoptysis and pulmonary hemorrhage, and the appropriate investigations to aid in determining the correct diagnosis. The management and prognosis of idiopathic pulmonary hemosiderosis, based on cumulative experience from published reports, are considered in more detail. PMID- 15114548 TI - Dynamic behavior of respiratory system during nasal continuous positive airway pressure in spontaneously breathing premature newborn infants. AB - The end-expiratory lung-volume level of premature newborn infants is maintained above passive resting volume during active breathing, through the combination of reduced time constant and high respiratory rate. To determine whether nasal continuous positive airway pressure (NCPAP) alters this characteristic dynamic breathing pattern, we studied the effects of various NCPAP levels on the dynamic elevation of end-expiratory lung volume level (DeltaEELV) in spontaneously breathing premature newborn infants, using respiratory inductive plethysmography (RIP). Eleven premature newborn infants with moderate respiratory failure were included. NCPAP levels were set in a random order to 0, 2, 4, and 6 cm H2O. Tidal volume (Vt), rib-cage contribution to Vt (%RC), phase angle between abdominal and thoracic motions (theta), respiratory rate (RR), and inspiratory and expiratory times (Ti and Te) were continuously recorded by RIP. The slope of the linear part of the expiratory flow-volume relation was extrapolated up to zero flow level to evaluate the dynamic elevation of the functional residual capacity (FRC) (DeltaEELV). The time-constant of the respiratory system (tauRS) was calculated as the slope of the linear part of the expiratory flow-volume loop. At NCPAP = 6 cm H2O, DeltaEELV reached 0.6 +/- 0.2 times the Vt at NCPAP = 0 cm H2O. An increase in NCPAP level resulted in a significant decrease in DeltaEELV (P < 0.01). A decrease in DeltaEELV during NCPAP was associated with a significant increase in Te from 0.62 +/- 0.13 sec at NCPAP = 0 cm H2O to 0.80 +/- 0.07 sec at NCPAP = 6 cm H2O (P < 0.05), and a decrease in tauRS from 0.4 +/- 0.1 sec at NCPAP = 0 cm H2O to 0.24 +/- 0.04 sec at NCPAP = 6 cm H2O (P < 0.01). These results indicate that the characteristic spontaneous breathing pattern causing a dynamic elevation of FRC is abolished by NCPAP. We speculate that the dynamic volume-preserving mechanisms resulting from expiratory flow braking are no longer required during NCPAP, as the constant pressure may passively elevate FRC. PMID- 15114549 TI - Tobacco smoke exposure, wheeze, and atopy. AB - We investigated the effect of in utero and postnatal environmental tobacco smoke (ETS) exposure on respiratory symptoms and atopy in the first 3 years of life in children at high risk of allergic disease (both parents atopic). Three hundred and sixty-nine children were followed from birth and reviewed at ages 1 and 3 years (respiratory questionnaire, skin testing). Parental smoking questionnaires were administered, and plasma cotinine in cord and peripheral blood (at age 1 year) was measured (capillary column gas-liquid chromatography). Wheezing starting in the first year of life was significantly more common in children of smoking mothers (54.2% vs. 39.5%, P = 0.017), but not wheezing starting after age 1 year (10.8% vs. 10.9%, smoking and nonsmoking mothers, P = 0.99). Detectable cord cotinine was not associated with wheeze. More frequent wheeze in infancy was significantly more common in those with detectable 1-year cotinine (e.g., wheeze without colds, 17.8% vs. 5.6%, P = 0.02; wheeze most days, 6.5% vs. 0%, P = 0.04). ETS exposure was not associated with atopy. In the multivariate regression analysis, maternal smoking during pregnancy and/or in the first year of life remained associated with wheeze in the first year of life (odds ratio, 1.88; 95% confidence interval, 1.14-3.12; P = 0.01). ETS exposure in "high-risk" infants increases the risk of wheezing starting in the first year of life, but not after age 1 year. However, ETS exposure has little or no effect on the development of atopy. Measurement of plasma cotinine was no more useful than tobacco exposure assessment by questionnaire in our cohort. PMID- 15114551 TI - Impact of targeted-volume ventilation on lung inflammatory response in preterm infants with respiratory distress syndrome (RDS). AB - Volutrauma and pulmonary inflammation are thought to be the most important predisposing factors of chronic lung disease (CLD), a major complication of prematurity. A new option in patient-triggered ventilation (PTV), the volume guarantee (VG), a volume-targeted ventilation, seems to be a promising approach in reducing the risk of CLD, by limiting lung inflammatory injury and volutrauma. Our aim was to evaluate lung inflammatory response in preterm infants with respiratory distress syndrome (RDS), mechanically ventilated with and without VG, as measured by proinflammatory cytokines (IL-6, IL-8, and TNF-alpha) in tracheobronchial aspirate (TA) fluid. Fifty-three preterm infants (GA = 25-32 weeks) with RDS were randomized at birth to be ventilated using pressure support ventilation (PSV) with VG (Vt = 5 ml/kg) (n = 30) and without VG (n = 23) (Draeger Babylog 8000 Plus, 5.n). IL-6, IL-8, and TNF-alpha were determined by ELISA in TA samples on days 1, 3, and 7 of life. We observed a significant difference (ANOVA) in IL-8 and IL-6 levels on day 3 between the two groups (P < 0.05), and an increasing significative trend in IL-8 values in PSV group (P < 0.05). Mechanical ventilation lasted longer in the PSV group (12.3 +/- 3 vs. 8.8 +/- 3 days) (P = no significance). In conclusion, these preliminary data suggest a role for volume-targeted ventilatory strategy in reducing acute inflammatory response in preterm infants with RDS. Further studies are required in order to define whether this ventilatory strategy prevents lung injury. PMID- 15114550 TI - Sleep-disordered breathing in 3,680 Greek children. AB - The goal of the present investigation was to describe the prevalence of and clinical factors associated with sleep-disordered breathing in children and adolescents. Children and adolescents (3,680 in all, 1-18 years old) attending schools in central Greece were surveyed by questionnaires distributed to parents. We found a similar prevalence of habitual snoring (present every night) among three different age groups (5.3%, 4%, and 3.8% in 1-6-, 7-12-, and 13-18-year-old subjects, P = NS). Several children with an adenoidectomy and/or tonsillectomy were snoring every night (6.1%), whereas sleepiness at school was more common in habitual snorers than in nonhabitual snorers (4.6 vs. 2%, P = 0.03). Seventy randomly selected subjects among 307 snorers without adenoidectomy and/or tonsillectomy underwent polysomnography. The estimated frequency of obstructive sleep apnea-hypopnea among children without adenoidectomy and/or tonsillectomy was 4.3%. Factors associated with snoring were: male gender (odds ratio 1.5 (confidence interval, 1.2-1.9)); chronic rhinitis (2.1 (1.6-2.7)); snoring in father (1.5 (1.2-1.9)), mother (1.5 (1.1-2.0)), or siblings (1.7 (1.2-2.4)); adenoidectomy in mother (1.5 (1.0-2.2)); and passive smoking (1.4 (1.1-1.8)). In conclusion, snoring every night was equally prevalent in younger and older ages, more frequent in males, and present even in some children with a history of adenoidectomy and/or tonsillectomy. Chronic rhinitis, family history of snoring, and exposure to cigarette smoke were associated with an increased frequency of habitual snoring. PMID- 15114552 TI - Comparison of spirometric reference values. AB - Lung-function reference values play a vital role in the management of respiratory disorders. There are many proposed reference equations for pediatric spirometry. Recently, spirometric reference equations were proposed, using data from people aged 8-80 years living in the US compiled by the third National Health and Nutrition Examination Survey. Our objective was to compare the predictive value of wider age-range reference equations to established pediatric reference equations for the pediatric population. Spirometry, height, and weight were obtained from 70 normal children aged 6-18 years. The difference between measured and predicted values as suggested by different reference equations was compared. Predicted values from general equations significantly differed from those generated from pediatric equations and from measured values in this population. The difference between measured and predicted values from the wider age-range equations varied between 7-16% for forced expired volume in 1 sec (FEV1) and forced vital capacity (FVC). The difference between measured and predicted values for the pediatric equations varied between 1-4%. Although wider age-range equations provide continuity through age ranges, their predictive accuracy may be low in the pediatric age group, especially for the youngest, smallest children. Extrapolating reference equations beyond the age range of subjects used to generate then is not recommended. PMID- 15114553 TI - Sex differences among children 2-13 years of age presenting at the emergency department with acute asthma. AB - Hospitalization rates for asthma have been reported to be higher in males than females in children under age 15, but it is not clear whether this disparity reflects gender differences in prevalence, severity, or treatment. We performed a prospective cohort study as part of the Emergency Medicine Network. Patients aged 2-13 years who presented to the emergency department (ED) with acute asthma underwent a structured interview in the ED and another by telephone 2 weeks later. Of 1,602 patients, 61% (95% CI, 59-64%) were boys. Girls were slightly older than boys, although no material differences existed in acute presentation, chronic asthma characteristics, ED treatment, or ED course. There was no difference in admission rates for boys or girls (20% vs. 22%; P = 0.48). This finding persisted when adjusting for other factors in a multivariate logistic regression model. No sex differences were observed for relapse or ongoing exacerbation on univariate or multivariate analysis. These data suggest that asthma is not inherently more severe in boys with asthma compared to girls, and that the increased rate of hospitalizations in boys under age 13 is due to differences in prevalence, not severity. PMID- 15114554 TI - Urban air pollution and children's asthma: what do parents and health professionals think? AB - Our objective was to explore and compare, in the context of other exposures, lay and professional perceptions of the links between urban air pollution and children's asthma. We used a triangulated survey approach, using quantitative questionnaire surveys enriched by qualitative interviews. Derivation of indicators of actual local air quality used modelled air pollution and a geographical information system. Our setting involved families and community health professionals in the London borough of Ealing, and pediatric respiratory specialists across the United Kingdom. Participants included 863 parents of children aged 3-11 years, 151 reporting currently asthmatic children, of whom 20 were extensively interviewed; 98 local general practitioners and 50 practice nurses; and 75 paediatric respiratory consultants and 55 specialist nurses. Main outcome measures involved views about the links between urban air pollution and children's asthma, relative to other triggers. Comparison of assessments of local air quality, with actual pollution levels, was made by parents with and without asthmatic children. Many parents were unsure as to what factors initiate asthma, but the most frequently cited was traffic pollution; it was also considered important in the exacerbation of asthma. Health professionals' assessments were inconsistent: specialists conformed to the dominant literature dismissing strong links between air pollution and asthma, while local clinicians reflected the views of parents in their community. Surrounding parents' views were difficulties defining exposures to urban air pollution, underlying concerns about risks to general health, perceived lack of control, unclear expert opinion, and widely accepted informal "messages" which assumed strong links. Parents with experience of asthma were found to have significantly less accurate (negatively biased) perceptions of local air quality. In conclusion, reactions to uncertainty surrounding associations between asthma and urban air pollution varied: parents' concerns were heightened (and propagated by other influences), specialist clinicians were dismissive, and community clinicians fell between these extremes. PMID- 15114555 TI - Identification of Pseudomonas aeruginosa, Burkholderia cepacia complex, and Stenotrophomonas maltophilia in respiratory samples from cystic fibrosis patients using multiplex PCR. AB - A multiplex PCR method was developed to identify P. aeruginosa, B. cepacia complex, and S. maltophilia directly in sputum and oropharyngeal samples from CF patients. One hundred and six patients (53 male, and 53 female) attending our pulmonology clinic were studied from September 2000-April 2001. Two hundred and fifty-seven samples were cultured in selective media and submitted to multiplex PCR reactions, using three primer pairs targeting specific genomic sequences of each species, with an additional primer pair targeting a stretch of ribosomal 16S DNA, universal for bacteria, to act as a control. P. aeruginosa was isolated by culture in 56% of samples, B. cepacia complex in 4.3%, and S. maltophilia in 2.7%, while multiplex PCR identified P. aeruginosa in 78.7%, B. cepacia complex in 3.9%, and S. maltophilia in 3.1% of samples. Multiplex PCR results were verified by PCR reactions using different species-specific primers described in the literature and DNA sequencing of amplicons from a few samples. Comparing to culture results, the sensitivity and specificity values of multiplex PCR for bacterial identification were, respectively, 97.2% and 45.5% for P. aeruginosa, 45.5% and 97.9% for B. cepacia complex, and 40% and 97.6% for S. maltophilia. All 10 multiplex PCR-positive results for B. cepacia complex were confirmed using other species-specific primers described in the literature, while this approach confirmed results for S. maltophilia identification in 7/8 samples (87.5%). Sequencing of amplicons from samples culture-negative but multiplex PCR-positive for P. aeruginosa and B. cepacia complex confirmed their identity, while minor nucleotide differences among amplicons ruled out the hypothesis of PCR contamination. PMID- 15114556 TI - Assessment of airway function in young children with asthma: comparison of spirometry, interrupter technique, and tidal flow by inductance plethsmography. AB - The assessment of airway function in young children requires adaptation of techniques designed for adults and/or application of techniques that do not require complex respiratory maneuvers. We sought to assess two methods of measuring airway function: time to peak expiratory flows as a ratio of expiratory time (T(PTEF)/T(E)), derived from respiratory inductance plethysmography, and total respiratory resistance by the interrupter technique (Rint), both obtained during quiet tidal breathing. Both techniques were referenced to FEV1 and flow at 50% expired volume (FEF50) from conventional spirometry in 30 children aged 4-8 years (median age, 6.9; range, 4.5-8.5 years) with a physician diagnosis of asthma and who were able to perform FEV1 with a repeatability of at least 8%. T(PTEF)/T(E) and Rint were performed in random order followed by spirometry, in order to reduce the possible effects of pulmonary stretch on tidal breathing measures. Coefficients of variation (CV) and mean absolute change/baseline standard deviation were derived for each measurement. Baseline FEV1 did not correlate significantly with T(PTEF)/T(E) (r = 0.025), but did correlate with Rint (r = 0.737, P < 0.001); respective relationships for change after bronchodilator were r = 0.09 (ns) and r = 0.64 (P < 0.001). FEF50 also correlated significantly with Rint (R = 0.769, P < 0.001) but not with T(PTEF)/T(E). FEV1 and FEF50 both increased postbronchodilator, with respective mean changes of 11.4% and 28% (P < 0.001), while Rint decreased by 24.3% (P < 0.001). No significant changes were noted for T(PTEF)/T(E). T(PTEF)/T(E) derived from inductance plethysmography does not detect mild airway obstruction or modest changes in airway caliber following bronchodilator in young children with asthma. The interrupter technique may have a role in assessing baseline airway function and response to therapy in children unable to perform reliable spirometry, and/or when the investigator wishes to avoid the possible influence of forced maneuvers on airway tone. PMID- 15114557 TI - Single-breath exhaled nitric oxide in preschool children facilitated by a servo controlled device maintaining constant flow. AB - Fractional concentration of exhaled nitric oxide (FENO), an index of airway inflammation, is optimally measured in adults and school-age children using a single-breath online (SBOL) exhalation at constant flow. However, preschool-aged (<6 years old) children have difficulty exhaling at constant flow, and alternative methods are needed. We employed a servo-controlled variable resistance device (servo device) that controls expiratory flow while allowing the child to vary expiratory pressure. To validate this device, 8 children (aged 6-12 years) performed SBOL exhalations with and without the servo device at expired flow rates between 20-50 ml/sec. We then studied 32 young children aged 24-71 months with the servo device alone at exhalation flows of 30, 40, and 50 ml/sec. Test difficulty (TD) with each method was rated by questioning the older children, or as observed by the physician obtaining the data in the younger children (0 = no difficulty, 1 = mild difficulty, 2 = moderate difficulty, and 3 = unable to perform test). In the older children, SBOL exhalations with and without the servo device demonstrated equivalent flow-dependence of FENO values. Test difficulty was low (0.125-0.625) at all flow rates, with excellent agreement between the two methods (P < 0.001). Twenty-eight young children (<6 years old) were able to complete measurements at all three flow rates evaluated. The 4 subjects who were not able to successfully complete all the measurements were between 2-3 years old (mean 2.75 +/- SD). Exhaled NO (mean +/- SD; ppb) was 8.8 (+/-6.2), 10.6 (+/-6.7), and 13.2 (+/-8.8) ppb at flows of 50 ml/sec, 40 ml/sec, and 30 ml/sec, respectively. Mean values of SD scores were 1.00, 1.14, and 1.43 at flows of 50, 40, and 30 ml/sec, respectively (P = NS). In conclusion, exhaled NO measurement by the SBOL method was facilitated in preschool children by the use of a servo-controlled variable resistance device. This device may allow these measurements to be applied to aid in the diagnosis and treatment of asthma in the preschool child, where spirometry is generally impossible. PMID- 15114563 TI - Molecular peapods as supramolecular carbon allotropes. PMID- 15114558 TI - Cytomegalovirus pneumonitis in a patient with Hodgkin disease and ataxia telangiectasia. AB - We describe a child with with Hodgkin disease and ataxia-telangiectasia who also developed an unusual pneumonitis caused by a cytomegalovirus which was fatal. PMID- 15114564 TI - Chemical safety in a vulnerable world--a manifesto. PMID- 15114565 TI - Functional porous coordination polymers. AB - The chemistry of the coordination polymers has in recent years advanced extensively, affording various architectures, which are constructed from a variety of molecular building blocks with different interactions between them. The next challenge is the chemical and physical functionalization of these architectures, through the porous properties of the frameworks. This review concentrates on three aspects of coordination polymers: 1). the use of crystal engineering to construct porous frameworks from connectors and linkers ("nanospace engineering"), 2). characterizing and cataloging the porous properties by functions for storage, exchange, separation, etc., and 3). the next generation of porous functions based on dynamic crystal transformations caused by guest molecules or physical stimuli. Our aim is to present the state of the art chemistry and physics of and in the micropores of porous coordination polymers. PMID- 15114566 TI - Stereoisomeric discrimination in DNA compaction. PMID- 15114568 TI - Self-assembly of two distinct supramolecular motifs in a single crystalline framework. PMID- 15114567 TI - Highly enantioselective phase-transfer-catalytic alkylation of 2-phenyl-2 oxazoline-4-carboxylic acid tert-butyl ester for the asymmetric synthesis of alpha-alkyl serines. PMID- 15114569 TI - Fluorescent probes for hydrogen peroxide based on a non-oxidative mechanism. PMID- 15114570 TI - Light-driven machine prototypes based on dissociative excited states: photoinduced decoordination and thermal recoordination of a ring in a ruthenium(II)-containing [2]catenane. PMID- 15114571 TI - Magnetic relaxation switch immunosensors detect enantiomeric impurities. PMID- 15114572 TI - Hydrogen-bonded helices in the layered aluminophosphate (C2H8N)2[Al2(HPO4)(PO4)2]. PMID- 15114573 TI - Cationic gold(I) complexes: highly alkynophilic catalysts for the exo- and endo cyclization of enynes. PMID- 15114574 TI - Two-color two-laser DNA damaging. PMID- 15114575 TI - A mixed-valence tin-oxygen cluster containing six peripheral ferrocene units. PMID- 15114576 TI - Origin of diastereoselection in the hydrosilylation of chiral N-acyliminium intermediates derived from pyroglutamic acid. PMID- 15114577 TI - Desulfurization of flue gas: SO(2) absorption by an ionic liquid. PMID- 15114578 TI - Direct evidence for oxygen-atom exchange between nonheme oxoiron(IV) complexes and isotopically labeled water. PMID- 15114579 TI - Synthesis of beta-hydroxyaldehydes with stereogenic quaternary carbon centers by direct organocatalytic asymmetric aldol reactions. PMID- 15114580 TI - Cascade carbonylation methods leading to beta-diketones and beta-functionalized delta-diketones. PMID- 15114581 TI - A highly effective phosphoramidite ligand for asymmetric allylic substitution. PMID- 15114582 TI - Nickel-catalyzed cross-coupling of aryl grignard reagents with aromatic alkyl ethers: an efficient synthesis of unsymmetrical biaryls. PMID- 15114583 TI - Pseudodynamic combinatorial libraries: a receptor-assisted approach for drug discovery. PMID- 15114584 TI - Arylnaphthalene lignans through Pd-Catalyzed [2+2+2] cocyclization of arynes and diynes: total synthesis of Taiwanins C and E. PMID- 15114585 TI - Synthesis of amines from imines in the coordination sphere of silicon-surprising photo-rearrangement of hexacoordinate organosilanes. PMID- 15114586 TI - Metallocene-catalyzed C7-linkage in the hydrooligomerization of norbornene by sigma-bond metathesis: insight into the microstructure of polynorbornene. PMID- 15114587 TI - Highly modular construction of differently substituted dihydrodibenzo[a,c]cycloheptenes: fast and efficient access to derivatives of 2,2'-cyclo-7,8'-neolignans. PMID- 15114589 TI - Predictive value of QT dispersion for acute heart failure after autologous and allogeneic hematopoietic stem cell transplantation. AB - The aim of our study was to evaluate whether corrected QT dispersion (QTc dispersion), an electrocardiographic marker, is a good predictor of the development of acute heart failure after high-dose chemotherapy followed by autologous or allogeneic hematopoietic stem cell transplantation. We enrolled 50 consecutive patients, from age 15 to 63 years, with hematopoietic diseases scheduled to undergo autologous or allogeneic hematopoietic stem cell transplantation, and compared QTc dispersion with other markers before transplantation conditioning. In univariate logistic analysis, QTc dispersion was a significant factor for acute heart failure after hematopoietic stem cell transplantation (odds ratio, 3.7 per 10 msec; confidence interval, 1.6-8.5; P = 0.002). There were no significant differences as age, sex, systolic or diastolic echocardiographic function markers, cumulative anthracycline dose, or QTc before transplantation between patients with and without acute heart failure. After multiple adjustments for left ventricular ejection fraction, cumulative anthracycline dose, cyclophosphamide conditioning dose, QTc dispersion was a significant and independent factor for acute heart failure after hematopoietic stem cell transplantation (odds ratio, 48.0 per 10 msec; confidence interval, 1.4 1666.3; P = 0.03). This study demonstrated that QTc dispersion could be used as a powerful noninvasive predictor of the development of acute heart failure after hematopoietic stem cell transplantation. PMID- 15114590 TI - R147W mutation of PROC gene is common in venous thrombotic patients in Taiwanese Chinese. AB - We analyzed the genetic defects of 21 unrelated patients with venous thrombosis in whom hereditary protein C deficiency was diagnosed. Eleven mutations were detected in 18 families, while no mutation was detectable in the other three families. Among these mutations, a common genetic mutation of protein C (PROC) gene recurred in 43% (nine propositi from these 21 families). This C6152T mutation at exon 7 resulted in a missense mutation, Arg147Trp (R147W). Each propositus or family carrier had another specific polymorphism T66C at exon 2, which did not change the proline at position (-21). The haplotype analysis strongly suggested a founder effect. The first thrombotic attack was significantly younger in patients with multiple genetic defects or combined with other risk factors than those without precipitating factors. We further studied the prevalence rate of R147W mutations in normal populations through PCR amplification and dot hybridization. Three different cohorts got similar results. The prevalence rate of R147W in health controls is 0.85% [95% confidence interval (CI) 0.35-1.35]. Analysis of plasma protein C levels of all patients or carriers suggested that R147W was a type II deficiency. The odds ratio of thrombosis of R147W is 5.1 (95% CI 1.7-14.8). Taken together, R147W mutation is a significant thrombotic risk factor and is the most common defect of PROC gene in Taiwanese patients with protein C deficiency. This finding is important for screening thrombophilic families in Chinese populations. PMID- 15114591 TI - Cytotoxic T-lymphocyte antigen-4 (CTLA-4) gene polymorphism and susceptibility to non-Hodgkin's lymphoma. AB - The CTLA-4 molecule plays an important role in immune regulation by downregulating activation of T cells. Polymorphisms in the CTLA-4 gene have been shown to be associated to a number of autoimmune diseases including blood disorders. In this study, the intragenic polymorphisms of the CTLA-4 gene at position -318*C/T, +49*A/G, and the dinucleotide (AT)(n) repeat polymorphism in exon 3 were analyzed in patients with non-Hodgkin's lymphoma. Genotype and haplotype analysis showed that the exon 1+49*AA genotype was over-represented among patients with NHL (P = 0.002), whereas no difference was observed for the 318*C/T promoter and the (AT)(n) polymorphisms (P > 0.05). The data obtained indicate that the CTLA-4+49A/G polymorphism may have a role in genetic susceptibility to NHL. PMID- 15114592 TI - Effect of red cell exchange transfusion on plasma levels of inflammatory mediators in sickle cell patients with acute chest syndrome. AB - Red cell exchange transfusion is the recommended therapy for patients with sickle cell disease (SCD) who have severe, progressive acute chest syndrome (ACS). A double-volume red cell exchange transfusion decreases the percentage of hemoglobin S (Hgb S) containing red blood cells to less than 20%, improving vascular perfusion. We speculated that reduction of pro-inflammatory mediators might also contribute to the therapeutic effect of an exchange transfusion. We measured white blood cell count (WBC), absolute neutrophil count (ANC), platelet concentration as well as plasma levels of interleukin-1alpha (IL-1alpha), interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8), and soluble vascular cell adhesion molecule-1 (sVCAM-1) in 8 sickle cell patients with 9 episodes of ACS who received a manual, double volume exchange transfusion. Six patients with SCD seen during a routine clinic visit were used as controls. The mean number of hospitalization days was 6, with an average of 2 days in the intensive care unit. All patients recovered without complication. Sickle cell patients with ACS had a higher WBC and ANC at baseline but lower sVCAM-1 levels compared to controls. TNF-alpha, IL-1alpha, IL-1beta, and IL-8 levels were not significantly different from controls. WBC, ANC, platelet, and sVCAM-1 measurements were significantly decreased immediately post exchange in patients with ACS; however, this effect was not persistent as levels trended towards pre-exchange values by 24 hr post-exchange. Due to wide inter individual variability, a consistent pattern was not seen for TNF-alpha, IL 1alpha, IL-1beta, or IL-8. We conclude that in sickle cell patients with ACS, a manual, double-volume exchange transfusion lowers WBC, ANC, platelets, and sVCAM 1 levels, but the effect is short-lived. PMID- 15114593 TI - Protein C and S and inflammation in sickle cell disease. AB - Reduced activity of naturally occurring anticoagulants (NOAC) protein C and protein S may contribute to vaso-occlusion in sickle cell disease (SCD). We studied whether protein C and S are related to clinical vaso-occlusion, hematological markers of disease severity (hemoglobin levels, leukocyte counts, and percentage of fetal hemoglobin), and inflammation in SCD. Protein C activity, protein S (free and total) antigen, endothelial activation markers (soluble vascular cell adhesion molecule-1 [sVCAM-1], von Willebrand antigen [vWF]), and high sensitive C-reactive protein (hsCRP) levels were measured in 30 HbSS and 20 HbSC patients and in race-matched HbAA controls. NOAC levels were reduced in patients, and endothelial activation markers and hsCRP were elevated (except vWF in HbSC patients). Protein C activity and vWF levels were lower in HbSC patients who experienced painful crises compared to HbSC patients who were clinically asymptomatic. No other differences were observed between patients who did and did not experience vaso-occlusive events (painful crises, stroke, acute chest syndromes) or leg ulcers. A significant positive correlation between total protein S with hemoglobin levels and a significant negative correlation between total and free protein S and sVCAM-1 were detected in HbSS patients. Except perhaps for protein C in relation to painful crises in HbSC patients, these markers were not associated with the occurrence of clinical events. The protein S, hemoglobin, and sVCAM-1 associations may suggest decreased endothelial protein S production due to the more severe endothelial perturbation in HbSS patients with lower hemoglobin levels. PMID- 15114594 TI - Allogeneic hematopoietic stem cell transplantation for seven children with X linked hyper-IgM syndrome: a single center experience. AB - X-linked hyper-IgM syndrome (XHIM), or hyper-IgM syndrome type 1 (HIGM1), is a rare primary immunodeficiency disorder susceptible to recurrent bacterial infection and opportunistic infection such as Pneumocystis carinii and Cryptosporidium parvum. The long-term outcome is quite poor, and allogeneic hematopoietic stem cell transplantation (HSCT) offers the only cure. Seven patients with XHIM, from age 3 to 19 years (mean 11.3 years), underwent allogeneic HSCT in our institution. Details of pre- and post-transplantation data and transplantation procedure were analyzed retrospectively. The donors were HLA identical siblings for three patients and HLA-identical unrelated donors for four patients. All but one received conventional conditioning regimen consisting of busulfan and cyclophosphamide and prophylaxis for graft-versus-host disease (GVHD) consisting of cyclosporine and methotrexate. Five out of seven patients are alive and well with normal CD40L expression, and four of these five are free of intravenous immunoglobulin supplementation. The two patients who died had prolonged episodes of severe and recurrent infections and organ damage. We conclude that conventional allogeneic HSCT from HLA matched related or unrelated donors is curative and feasible for XHIM patients, if performed before significant infections and organ damage occur. For the high-risk patients, an alternative approach including nonmyeloablative HSCT may be more feasible. PMID- 15114595 TI - MTHFR C677T polymorphism and its relation to ischemic stroke in the Black Sea Turkish population. AB - The MTHFR C677T mutation has been shown to be associated with venous thrombosis. The role of this mutation in ischemic stroke is unclear. We investigated whether the MTHFR mutation is a risk factor for patients with ischemic stroke in the Black Sea Turkish population or not. We analyzed 30 patients (19 male, 11 female) [median age: 50 years (range: 28-78)] with ischemic stroke who had no known predisposition factors for stroke and 242 (182 male, 60 female) healthy controls [median age: 42 years (range: 18-65)]. Detection of the MTHFR C677T mutation was performed by using commercially available allele-specific PCR-ELISA kits. Prevalence of the MTHFR C677T genotype was 49.1% (CT, 45.8%; TT, 3.3%) in controls and 50% (CT, 43.3%; TT, 6.6%) in patients [OR: 1.03, 95% CI (0.45 2.35]). The prevalence of homozygous gene mutation for MTHFR was higher among patients with stroke than control subjects, but this difference was not statistically significant. The MTHFR gene mutation is not a risk factor for ischemic stroke formation in patients from the Black Sea region in Turkey. PMID- 15114596 TI - Microgranular and t(11;17)/PLZF-RARalpha variants of acute promyelocytic leukemia also present the flow cytometric pattern of CD13, CD34, and CD15 expression characteristic of PML-RARalpha gene rearrangement. AB - Acute promyelocytic leukemia (APL) is a subtype acute myeloid leukemia in which leukemic promyelocytes predominate in the bone marrow (BM). Rapid diagnosis is critical for treatment decision since all-trans-retinoic acid must be administrated promptly. The microgranular variant may be of difficult diagnosis, as it may be confused with other diseases on morphological grounds. The purpose of this study was to determine if the microgranular variant has the same antigenic profile as the classical hypergranular type. The immunophenotype of leukemic cells from the bone marrow of 50 patients, with the PML-RARalpha gene rearrangement confirmed by RT-PCR, was determined by flow cytometry using a large panel of 22 monoclonal antibodies and a polyclonal anti-TdT antibody. Thirty-four cases were classified as classical APL and 16 as microgranular APL. The immunophenotypic profile of the two subtypes was indistinguishable concerning the presence or absence of these antigens, including the absence of reactivity for the HLA-DR antigen. The simultaneous immunophenotypic combination of a unique major cell population, heterogeneous intensity of expression of CD13, and the typical pattern of CD15/CD34 expression were similarly present in the hypergranular and microgranular subtypes. Homogeneous expression of CD33 was observed in 76% of the classical APL cases and in 100% of the microgranular cases. Additionally, we have studied two cases of PLZF-RARalpha APL that also displayed the same immunophenotype described for classical APL. Thus, the immunophenotypic profile highly characteristic of the PML-RARalpha gene rearrangement was also observed in microgranular and PLZF-RARalpha variants of APL. PMID- 15114597 TI - Second malignant neoplasms in childhood acute lymphoblastic leukemia: primitive neuroectodermal tumor of the chest wall with germline p53 mutation as a second malignant neoplasm. AB - About 80% of children treated for acute lymphoblastic leukemia (ALL) will be long term survivors. Second malignant neoplasm (SMNs) are a devastating sequelae observed on these children, with an estimated cumulative risk of 2-3.3% fifteen years after diagnosis. Primitive neuroectodermal tumor of bone (PNET) is rarely observed as a SMN following treatment of childhood ALL. The authors described the occurrence of a chest wall PNET of the bone at the site of a central line placement associated with both germ-line and tumor cell p53 mutation in a 8-year old boy 1 year after completing therapy for standard risk ALL. A review of the literature of 25,051 children treated for ALL discovered 230 SMNs (0.99%), and only one case of PNET of the bone was noted among this group. The occurrence of a SMN in children treated for ALL is a rare event. Such an occurrence, in particular the development of an unusual SMN, should be evaluated for a germline p53 mutation. PMID- 15114598 TI - Elevated plasma sVCAM-1 levels in children with sickle cell disease: impact of chronic transfusion therapy. AB - Vascular cell adhesion molecule-1 (VCAM-1) has been implicated as being important in the pathophysiology of acute pain episodes (APE) and acute chest syndrome (ACS) of sickle cell disease (SCD). The frequency of these episodes is reduced by chronic transfusion therapy. The impact of chronic transfusion therapy on VCAM-1 expression is unknown. Soluble VCAM-1 (sVCAM-1) levels were measured in plasma using an ELISA assay (R&D Systems) in 61 patients with SCD (age range 1.5-20 years) and 12 normal controls (2.5-14 years). SCD patients included 20 with ACS, 14 with APE, 12 at well-child visits, and 15 receiving chronic transfusion therapy. Asymptomatic SCD patients had higher sVCAM-1 levels compared to normal subjects (P < 0.001). Levels of sVCAM-1 were further elevated during ACS (P < 0.001) and APE (P = 0.072) and returned to the asymptomatic range on resolution. Levels were significantly lower in transfused patients (P = 0.003) compared to asymptomatic SCD patients. Our findings of increased VCAM-1 expression during ACS and perhaps APE offer a rationale for therapeutic use of cytokine and other VCAM 1 modulators. The reduction of sVCAM-1 levels observed in our transfused SCD patients offers insight into the mechanism of the protective effect of transfusion against ACS and APE and possibly stroke. PMID- 15114599 TI - Neutrophilic panniculitis with myelodysplastic syndromes presenting as pustulosis: case report and review of the literature. AB - Neutrophilic panniculitis associated with myelodysplastic syndromes is rare. We report a 59-year-old patient who initially was diagnosed with myelodysplastic syndrome (MDS) and developed a sudden onset of widespread pustulosis and erythematous indurated papules. Examination of skin biopsies of a papule lesion showed dense neutrophilic infiltration limited to the subcutaneous tissue. The pustules and papules disappeared completely after treatment with systemic corticosteroids. To our knowledge, only one patient was identified by MEDLINE search of the English-language literature. PMID- 15114600 TI - Thalidomide in POEMS syndrome: case report. AB - We report a patient with incapacitating POEMS syndrome characterized by serum monoclonal protein, polyneuropathy, organomegaly, endocrinopathy, mesangiocapillary glomerulonephritis, massive ascites formation, and pulmonary hypertension. A dramatic improvement in the clinical condition occurred after administration of thalidomide, a drug with known anti-angiogenetic, anti proliferative, and anti-cytokine properties. PMID- 15114601 TI - Thrombocytopenia due to acute venous thromboembolism and its role in expanding the differential diagnosis of heparin-induced thrombocytopenia. AB - Thrombocytopenia is an uncommon but serious consequence of heparin administration. Occasionally patients with massive acute venous thromboembolism (VTE) will develop thrombocytopenia. As heparin or some thrombin inhibitor is strongly indicated in acute VTE, it is important to distinguish this event from heparin-induced thrombocytopenia (HIT). Four patients are presented who developed thrombocytopenia so early in their course of VTE and/or therapy with heparin that HIT was considered unlikely. The mean nadir platelet count for these four patients was 60,000/microl occurring at a mean time of 18 hr after the initiation of heparin therapy. Because of strong indications to continue heparin for their acute VTE in the face of a very low likelihood that they did have HIT, heparin was continued with excellent results and resolution of the thrombocytopenia. The literature of this subject is reviewed. Thrombocytopenia following VTE is actually rather common, but it is usually milder than in these four cases. In some cases such as these four, the thrombocytopenia can be sudden and rather severe causing diagnostic confusion with HIT. PMID- 15114602 TI - Parenteral iron therapy options. AB - Parenteral iron therapy is occasionally necessary for patients intolerant or unresponsive to oral iron therapy, for receiving recombinant erythropoietin therapy, or for use in treating functional iron deficiency. There are now three parenteral iron products available: iron dextran, ferric gluconate, and iron sucrose. We summarize the advantages and disadvantages of each product, including risk of anaphylaxis and hypersensitivity, dosage regimens, and costs. The increased availability of multiple parenteral iron preparations should decrease the need to use red cell transfusions in patients with iron-deficiency anemia. PMID- 15114603 TI - Bone marrow findings in hereditary cystinosis with renal failure. PMID- 15114604 TI - Paucity of TEL-AML 1 translocation, by multiplex RT-PCR, in B-lineage acute lymphoblastic leukemia (ALL) in Indian patients. AB - A total of 69 patients of B lineage ALL, 35 children (32 males, 3 females) and 34 young adults (27 males, 7 females) were studied by multiplex RT-PCR to determine the relative frequency of t(9;22), t(12;21), t(1;19), and t(4;11,). Translocation (9;22) was seen in 1/35 (2.8%) and t(1;19) in 2/35 (5.7%) children. None of the children showed t(12;21) and t(4;11) translocations. In young adults, t(9;22) and t(1;19) were seen in 5/34 (14.7%) and 2/34 (5.8%) patients, respectively. None of the latter showed t(12;21) or t(4;11) translocations. Thus, there appears to be a significant under representation of the fusion transcripts for TEL-AML, a good prognostic marker, in this study, unlike in the West, where it is seen in 35% of children with ALL. This, together with the generally increased leukemic burden seen in Indian patients, may explain in part, the poor treatment outcome reported. PMID- 15114605 TI - Embolization of intracranial aneurysms and sickle cell disease. AB - The association of intracranial aneurysms and sickle cell disease (SCD) has been described in the English-language literature. Treatment strategies have included angiography and craniotomy. However, with advancement of non-operative interventions, much controversy surrounds the treatment of these patients. Endovascular embolization has been used in many patients with cerebral aneurysms, but it had been reported in the literature in only two sickle cell patients. The authors present a case of endovascular embolization of multiple intracranial aneurysms in a patient with sickle cell anemia with good outcome. PMID- 15114606 TI - Differences in the frequency of the C677T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene among the Lebanese population. AB - In view of its role in precipitating mild hyperhomocysteinemia as well as being a risk factor for vascular thrombosis, we investigated the frequency of the C677T mutation of the methylenetetrahydrofolate reductase (MTHFR) gene among 589 healthy Lebanese subjects by PCR-RFLP analysis (HinfI digestion) and compared them with those of other countries of Caucasian and non-Caucasian origin. The prevalence of the mutated homozygous (T/T) and heterozygous (C/T) C677T MTHFR genotype was 11.04% and 39.73%, respectively, giving an allele frequency of 0.309. While the prevalence of the T/T genotype was similar with respect to gender, higher prevalence was noted among Christian (13.08%) compared to Moslem (7.66%) subjects (P < 0.001), and heterogeneity in its distribution was seen in the different Lebanese provinces, and was directly related to the Christian/Moslem composition of each province. The distribution of the MTHFR C677T in Lebanon is unique with regard to its higher occurrence among Christians compared to Moslems, adding to the existing body of literature on the heterogeneity of its prevalence and distribution. PMID- 15114607 TI - Human herpesvirus 8 (HHV-8)-associated peritoneal primary effusion lymphoma (PEL) in two HIV-negative elderly patients. AB - Human herpesvirus 8 (KSHV/HHV-8) is associated with all forms of Kaposi sarcoma (KS), with a rare high-grade B-cell non-Hodgkin lymphoma characterized by serous effusions in body cavities called primary effusion lymphoma (PEL) and with some forms of multicentric Castleman disease (MCD). Although mostly observed during AIDS, such disorders have also been described with a lower incidence in human immunodeficiency virus-negative patients. We describe here the features of two novel cases of AIDS-unrelated PEL. Two patients, a 78-year-old man (case 1) and a 86-year-old woman (case 2), both of French origin, presented exudative ascitic effusion containing numerous KSHV/HHV-8(+) EBV(-) large lymphomatous cells of B cell clonal origin, characterized by a CD45(+) CD30(+) CD19(-) CD20(-) immunophenotype. The PEL tumor cells harbored a homogenous and isolated trisomy 12 in case 1 and an aberrant expression of the T-cell lineage antigen CD7 in case 2. Both patients were lymphopenic at the time of PEL diagnosis and rapidly died with progressive lymphoma. Moreover, patient 2 had a previous history of classic KS and MCD clinically improved after treatment with all-trans-retinoid acid and a concomitant metastatic breast adenocarcinoma. Compared to AIDS-related PEL, these two cases displayed distinct features in particular the advanced age of patients, as observed for Mediterranean KS, and the absence of EBV coinfection. PMID- 15114608 TI - Clinical spectrum of myelophthisis in cancer patients. AB - Myelophthisis is a form of bone marrow failure due to replacement of hematopoietic tissue by abnormal tissue, most commonly metastatic carcinomas. This results in extramedullary hematopoiesis, typically in the spleen leading to premature release of hematopoietic cells into the circulation. Peripheral blood findings may include nucleated red blood cells, tear drop forms, giant platelets, and immature leukocytes. This is called a leukoerythroblastic picture. The first case demonstrates acute myelophthisis as a presentation of pancreatic cancer. The second case is of extramedullary hematopoiesis as a manifestation of widely metastatic melanoma. The presence of a leukoerythroblastic peripheral blood picture should serve as a valuable clue about a possible underlying malignancy. This late presentation of advanced cancer may now be rarely seen because of early diagnosis and more effective therapies. PMID- 15114609 TI - Eosinophilia presenting as acute coronary syndrome. PMID- 15114610 TI - Isolated granulocytic sarcoma of the head and neck preceding acute myeloid leukemia. PMID- 15114611 TI - High incidence of acute promyelocytic leukemia in the Caucasian population: the Costa Rica experience. PMID- 15114612 TI - Major disorganization of factor VIII gene as a cause of severe hemophilia A in Indian patients. PMID- 15114613 TI - Low plasma level of all-trans retinoic acid after feeding tube administration for acute promyelocytic leukemia. PMID- 15114615 TI - Role of calcium and kinases on the neurotrophic effect induced by gamma aminobutyric acid. AB - An increasing body of evidence supports a trophic action of gamma-aminobutyric acid (GABA) during nervous system development. The purported mediator of these trophic effects is a depolarizing response triggered by GABA, which elicits a calcium influx in immature CNS cells. This Mini-Review focuses on the neurotrophic role of neural activity and GABA and some of the most common intracellular cascades activated by depolarization and trophic factors. Several biological effects induced by GABA in the developing nervous system are reviewed, with particular emphasis on what is known about calcium-dependent neurotrophic effects induced by GABA and its intracellular mechanisms. PMID- 15114616 TI - Olfactory receptors in the mouse septal organ. AB - In this study we have identified a repertoire of chemosensory receptors expressed in the septal organ (SO). The results suggest that septal organ neurons are specified to express receptor genes belonging to class II olfactory receptors that are also expressed in the main olfactory epithelium. We found no evidence for the expression of members from the vomeronasal receptor gene families. In the SO, no topography analogous to the receptor expression zones of the main olfactory epithelium was evident. The majority of identified receptors corresponds to genes with restricted expression in the medial and lateral zones of the main olfactory epithelium. This coincides with the expression of olfactory cell adhesion molecule (OCAM) throughout the SO, which is considered as a marker for the medial-lateral zones. In contrast, NADPH:quinone oxidoreductase 1 expression, a characteristic marker for the dorsal zone, was lacking in the SO. Most of the receptor types were found to be expressed in rather few SO neurons; as an exception, the receptor mOR244-3 was observed in a very high proportion of cells. Although a very high fraction of SO neurons expressed mOR244-3, we found no evidence for the coexpression of different receptors in individual cells. PMID- 15114617 TI - Implantation of dendritic cells in injured adult spinal cord results in activation of endogenous neural stem/progenitor cells leading to de novo neurogenesis and functional recovery. AB - We report a treatment for spinal cord injury involving implantation of dendritic cells (DCs), which act as antigen-presenting cells in the immune system. The novel mechanisms underlying this treatment produce functional recovery. Among the immune cells tested, DCs showed the strongest activity inducing proliferation and survival of neural stem/progenitor cells (NSPCs) in vitro. Furthermore, in DC implanted adult mice, endogenous NSPCs in the injured spinal cord were activated for mitotic de novo neurogenesis. These DCs produced neurotrophin-3 and activated endogenous microglia in the injured spinal cord. Behavioral analysis revealed the locomotor functions of DC-implanted mice to have recovered significantly as compared to those of control mice. Our results suggest that DC-implantation exerts trophic effects, including activation of endogenous NSPCs, leading to repair of the injured adult spinal cord. PMID- 15114618 TI - Rescue of TNFalpha-inhibited neuronal cells by IGF-1 involves Akt and c-Jun N terminal kinases. AB - Proinflammatory cytokines, especially tumor necrosis factor alpha (TNFalpha), is a pleiotropic mediator of a diverse array of physiologic and neurologic functions and is upregulated during various inflammatory and neurodegenerative diseases. A common survival response during such situations is the increased expression of the hormone insulin-like growth factor 1 (IGF-1). Although it was thought previously that the mechanisms of TNFalpha and IGF-1 action were unrelated, it has been shown that low doses of TNFalpha can inhibit the survival effects of IGF 1 in mouse cerebellar granule neurons. We used a neuronal cell line SH-SY5Y, which underwent apoptosis in response to TNFalpha and this process could be reversed substantially by IGF-1. Crosstalk between signaling pathways of these two factors was found at various points downstream of their signal transduction. To determine the mechanisms of IGF-1-mediated rescue, we looked at the MAP kinases, which are known to be involved in IGF-1 as well as TNFalpha signaling. The c-Jun N-terminal kinase pathway, which is known normally to promote cell death, was found to actually promote survival of TNFalpha-mediated cell death. Inhibiting the c-Jun survival pathway completely reversed the rescue mediated by IGF-1. In addition, the Akt pathway played an equally important role in this rescue. PMID- 15114619 TI - Input resistance is voltage dependent due to activation of Ih channels in rat CA1 pyramidal cells. AB - The contribution of the hyperpolarization-activated cation current (I(h)) to input resistance (R(N)) and resting potential (RP) was investigated during whole cell patch-clamp recordings in CA1 pyramidal cells of rat hippocampal slices. In current-clamp mode, R(N) was determined at different membrane potentials. R(N) decreased with increasing hyperpolarization, from about 260 Momega to 140 Momega at potentials of about -60 mV and -110 mV, respectively. Both the potential of half-maximal reduction of R(N) and the potential of half-maximal I(h) activation (determined in voltage-clamp mode) were approximately -90 mV. The analysis of the voltage sag indicative of I(h) activation revealed a preferential activity of I(h) channels in a voltage range between -70 and -95 mV. ZD7288 (50 microM), a specific I(h) blocker, led to a hyperpolarization by about 4.8 mV, increased R(N) by approximately 45% within a potential range between -65 and -80 mV, and abolished the voltage dependence of R(N). Gabapentin (GBP, 100 microM), an I(h) channel agonist, led to a depolarization by about 2.4 mV and reduced R(N) by about 20% within a potential range between -65 and -80 mV. In conclusion, our data show that R(N) is voltage dependent due to I(h) channel activation and that I(h) channels are preferentially active at voltages between -70 and -95 mV. Furthermore, we demonstrated that R(N) can be modulated by antiepileptic drugs such as GBP, which may partly explain its antiepileptic effect as due to decreasing the sensitivity to excitatory input. PMID- 15114620 TI - Dependence of calcium influx in neocortical cells on temporal structure of depolarization, number of spikes, and blockade of NMDA receptors. AB - Increase of intracellular [Ca(2+)] evoked by action potentials in a cell can induce long-term synaptic plasticity even without concomitant presynaptic stimulation. We used optical recording of the fluorescence of a Ca(2+)-indicator Oregon Green to investigate whether differences in results obtained with modifications of that purely postsynaptic induction protocol could be due to differential Ca(2+) influx. We compared changes of the somatic [Ca(2+)] in layer II-III pyramidal cells in slices of rat visual cortex evoked by bursts of depolarization pulses and long depolarizing steps. During weak depolarizations, the Ca(2+) influx was proportional to the amplitude and duration of the depolarization. With suprathreshold depolarizations, the Ca(2+) influx was proportional to the number of action potentials. Because the burst depolarizations evoked more spikes than did the long duration steps, this burst protocol led to a larger Ca(2+) influx. With all stimulation protocols, the spike induced Ca(2+) influx was reduced during blockade of N-methyl-D-aspartate (NMDA) receptors. Differences in intracellular [Ca(2+)] increases thus may be one reason for differential effects of purely postsynaptic challenges on synaptic transmission. PMID- 15114621 TI - Early adoption modifies the effects of prenatal stress on dopamine and glutamate receptors in adult rat brain. AB - Stressful stimuli during pregnancy induce complex effects that influence the development of offspring. These effects can be prevented by environmental manipulations during the early postnatal period. Repeated restraint during the last week of pregnancy was used as a model of prenatal stress, and adoption at birth was used to change the postnatal environment. No differences were found in various physical landmarks, except for testis descent, for which all prenatally stressed pups showed a 1-day delay in comparison with control rats, regardless of the postnatal adoption procedure. Levels of dopamine (DA) D(2) and glutamate (Glu) N-methyl-D-aspartate (NMDA) receptors were differentially regulated in different forebrain regions of cross-fostered adult offspring. Increased concentrations of cortical D(2) receptors detected in stressed pups, raised by a gestationally stressed biological mother, were not detected when the pups were raised by a control mother. Control pups raised by a foster mother whether gestationally stressed or not had higher levels of NMDA receptors in cortical areas. These findings suggest that the normal expression of DA and Glu receptors is influenced by in utero experience and by lactation. The complex pattern of receptor changes reflects the high vulnerability of DA and Glu systems to variations both in prenatal and in postnatal environment, particularly for cortical D(2) receptors and NMDA receptors in cerebral cortex and nucleus accumbens. In contrast, testis descent appears to be more susceptible to prenatal than to postnatal environmental events. PMID- 15114622 TI - Computer-identified nuclear localization signal in exon 1A of the transporter DMT1 is essentially ineffective in nuclear targeting. AB - Divalent metal transporter 1 (DMT1; also called DCT1, Nramp2, or SLC11A2) has multiple isoforms that localize differently in many cell types. DMT1 +IRE species (encoded by mRNA with an iron-responsive element) are limited to the plasma membrane and cytosolic vesicles. In neural cells, -IRE isoforms of DMT1 (encoded by mRNA lacking an IRE) localize to the nucleus, plasma membrane, and cytosolic vesicles. In considering nuclear compartmentalization of -IRE isoforms, we hypothesized that the newly identified exon 1A in the N-terminus of this transporter might contain a nuclear localization signal. DNA constructs starting with exon 1A and ending with exons encoding alternative isoforms were made and transiently transfected into HEK293T and PC12 cells as well as rat sympathetic neurons. None of the constructs appeared in the nucleus despite the presence of exon 1A. Antibody specific for exon 1A was also used in both immunostaining and Western blots to investigate localization of exon 1A expressed both endogenously and ectopically in cells. Again, nuclear localization of DMT1 containing exon 1A was not observed. Our data suggest that exon 1A is neither sufficient nor necessary for DMT1 to appear in the nucleus. PMID- 15114623 TI - Screening for control genes in rat global cerebral ischemia using high-density oligonucleotide array. AB - From conventional relative gene expression analyses (Northern blotting, in situ hybridization, and RT-PCR), it has been reported that the expression of control genes, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and beta-actin, used as references may be affected by ischemia. Therefore, we extended searching and evaluation at the mRNA level of transcripts whose expression levels were not changed by cerebral ischemia, using a high-density oligonucleotide array and statistical analysis in a rat global cerebral ischemia and reperfusion model. We added a hyperthermic factor and localization factor to ischemia and identified transcripts with a stable expression level under conditions even more disadvantageous than ischemia only. Screening of more than 8,000 transcripts with the Rat Genome U34A array yielded 28 transcripts, which we listed and classified according to their expression level. Widely used control genes, GAPDH and beta actin, were not included, although cyclophilin A was included. In addition, we conducted a functional classification based on gene ontology. Under the functional classification of the 28 transcripts, many genes tended to be associated with metabolism. In conclusion, use of several transcripts is recommended, such as those we identified, as references in the analysis of gene expression in pathological models of ischemia. PMID- 15114624 TI - Administration of N-acetylcysteine after focal cerebral ischemia protects brain and reduces inflammation in a rat model of experimental stroke. AB - Free radicals and inflammatory mediators are involved in transient focal cerebral ischemia (FCI). Preadministration of N-acetylcysteine (NAC) has been found to attenuate the cerebral ischemia-reperfusion injury in a rat model of experimental stroke. This study was undertaken to investigate the neuroprotective potential of NAC administered after ischemic events in experimental stroke. FCI was induced for 30 min by occluding the middle cerebral artery (MCA). NAC (150 mg/kg) was administered intraperitoneally at the time of reperfusion followed by another dose 6 hr later. Animals were sacrificed after 24 hr of reperfusion. The cerebral infarct consistently involved the cortex and striatum. Infarction was assessed by staining the brain sections with 2,3,5-triphenyltetrazolium chloride. Animals treated with NAC showed a significant reduction in infarct area and infarct volume and an improvement in neurologic scores and glutathione level. Reduction in infarction was significant even when a single dose of NAC was administered at 6 hr of reperfusion. Immunohistochemical and quantitative real-time PCR studies demonstrated a reduction in the expression of proinflammatory cytokines such as tumor necrosis factor alpha (TNFalpha) and interleukin 1beta (IL-1beta) and inducible nitric oxide synthase (iNOS) in NAC compared to that in vehicle-treated animals. The expression of activated macrophage/microglia (ED1) and apoptotic cell death in ischemic brain was also reduced by NAC treatment. These results indicate that in a rat model of experimental stroke, administration of NAC even after ischemia onset protected the brain from free radical injury, apoptosis, and inflammation, with a wide treatment window. PMID- 15114625 TI - Extracellular taurine in the substantia nigra: taurine-glutamate interaction. AB - Taurine has been proposed as an inhibitory transmitter in the substantia nigra (SN), but the mechanisms involved in its release and uptake remain practically unexplored. We studied the extracellular pool of taurine in the rat's SN by using microdialysis methods, paying particular attention to the taurine-glutamate (GLU) interaction. Extracellular taurine increased after cell depolarization with high K(+) in a Ca(2+)-dependent manner, being modified by the local perfusion of GLU, GLU receptor agonists, and zinc. Nigral administration of taurine increased the extracellular concentration of gamma-aminobutyric acid (GABA) and GLU, the transmitters of the two main inputs of the SN. The modification of the glial metabolism with fluocitrate and L-methionine sulfoximine also changed the extracellular concentration of taurine. The complex regulation of the extracellular pool of taurine, its interaction with GABA and GLU, and the involvement of glial cells in its regulation suggest a volume transmission role for taurine in the SN. PMID- 15114626 TI - Tyrosine hydroxylase and dopamine transporter expression following 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine-induced neurodegeneration of the mouse nigrostriatal pathway. AB - Administration of the neurotoxicant 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to C57BL/6 mice targets nigrostriatal dopaminergic neurons, leading to cell death and the depletion of striatal dopamine. After MPTP lesioning in young adult mice, surviving nigrostriatal dopaminergic neurons display robust and reproducible return of striatal dopamine weeks to months after injury. Thus, the mouse provides an excellent model with which to investigate the mechanisms underlying neuroplasticity of the nigrostriatal system following neurotoxic injury. The purpose of this study was to analyze proteins and mRNA transcripts of genes involved in dopamine biosynthesis (tyrosine hydroxylase; TH) and uptake (dopamine transporter; DAT) with regard to time course (7-90 days) after MPTP lesioning. Molecular analysis using immunohistochemistry and Western immunoblotting techniques demonstrated an increase in striatal TH by 30-60 days postlesioning that returned to near-control (prelesioned) levels by 60-90 days. In situ hybridization histochemistry indicated that this increase in TH protein might be due in part to increased TH mRNA expression in surviving nigrostriatal dopaminergic neurons. Analysis of TH protein at 7, 30, 60, and 90 days postlesioning with two-dimensional polyacrylamide gel electrophoresis in conjunction with Western immunoblotting revealed altered TH protein isoforms migrating at isoelectric points different from those of the native isoform. In contrast to TH protein, which returned to prelesioned levels by 60 days, DAT protein analysis showed that increased expression of striatal DAT protein did not return to near-prelesion levels until 90 days postlesioning. These results suggest that TH and DAT may differ in their time course of expression in surviving dopaminergic neurons and may play a role in mediating the return of striatal dopamine. PMID- 15114627 TI - Systemic administration of N-acetylcysteine protects dopaminergic neurons against 6-hydroxydopamine-induced degeneration. AB - The results of several in vitro studies have shown that cysteine prodrugs, particularly N-acetylcysteine, are effective antioxidants that increase the survival of dopaminergic neurons. N-acetylcysteine can be systemically administered to deliver cysteine to the brain and is of potential use for providing neuroprotection in the treatment of Parkinson's disease. However, it has also been reported that an excess of cysteine may induce neurotoxicity. In the present study, we injected adult rats intrastriatally with 2.5 microl of 6 hydroxydopamine (7.5 microg) and N-acetylcysteine (240 mM) or cysteine (240 mM) or intraventricularly with 6-hydroxydopamine (200 microg) and subcutaneously with N-acetylcysteine (10 and 100 mg/kg). We studied the effects of these compounds on both the nigrostriatal dopaminergic terminals and the surrounding striatal tissue. The tissue was stained with fluoro-jade (a marker of neuronal degeneration) and processed by immunohistochemistry to detect tyrosine hydroxylase, neuronal and glial markers, and the stress protein heme-oxygenase-1. After intrastriatal injection, both cysteine and N-acetylcysteine had clear neuroprotective effects on the striatal dopaminergic terminals, but also led to neuronal degeneration (as revealed by fluoro-jade staining) and astroglial and microglial activation, as well as intense induction of heme-oxygenase-1 in astrocytes and microglial cells. Subcutaneous administration of N-acetylcysteine also induced significant reduction of the dopaminergic lesion (about 30% reduction). However, we did not observe appreciable N-acetylcysteine-induced fluoro-jade labeling in striatal neurons or any of the above-mentioned changes in striatal glial cells. The results suggest that low doses of cysteine prodrugs may be useful neuroprotectors in the treatment of Parkinson's disease. PMID- 15114628 TI - 1-Benzyl-1,2,3,4-tetrahydroisoquinoline, a Parkinsonism-inducing endogenous toxin, increases alpha-synuclein expression and causes nuclear damage in human dopaminergic cells. AB - 1-Benzyl-1,2,3,4-tetrahydroisoquinoline (1BnTIQ), an endogenous neurotoxin, is known to cause parkinsonism in rodents and nonhuman primates. The levels of 1BnTIQ in cerebrospinal fluid of patients with Parkinson's disease (PD) were reported to be three times higher than those in control subjects. In the present study, we have evaluated the effects of 1BnTIQ on alpha-synuclein (alpha-syn) expression together with biochemical and morphological changes in human dopaminergic SH-SY5Y cells in culture. 1BnTIQ at lower concentrations (1-50 microM) increased alpha-syn protein expression in a time- and dose-dependent manner in these cells. There was also up-regulation of alpha-syn mRNA by 1BnTIQ. Inhibition of complex I by rotenone and depletion of glutathione by L-buthionine sulfoxamine also correlated with an increase in alpha-syn expression, suggesting that oxidative stress may cause an increase in alpha-syn levels in dopaminergic cells. Furthermore, 1BnTIQ significantly depleted glutathione levels. 1BnTIQ at higher concentrations (500 microM) increased reactive oxygen species levels, decreased ATP levels, and caused nuclear damage in the cells. The 1BnTIQ-induced alpha-syn up-regulation was inhibited by cotreatment with the antioxidants selegiline, coenzyme Q(10), and N-acetylcystein and the caspase inhibitor DEVD CHO. Taken together, these results suggest that alpha-syn up-regulation and oxidative stress are contributing factors in 1BnTIQ-induced neurotoxicity in dopaminergic neurons in PD. PMID- 15114629 TI - Progressive cognitive impairment and anxiety induction in the absence of plaque deposition in C57BL/6 inbred mice expressing transgenic amyloid precursor protein. AB - Numerous transgenic mouse models for Alzheimer's disease (AD) have been generated to recapitulate the histological pathogenesis and behavioral phenotypes of AD brain. However, none of the existing models exhibits the full spectrum of AD symptoms, nor have all of the traits mimicked by the developed animal models been successfully represented within a single mouse line, indicating that the development of transgenic lines showing new features of the AD-like brain should be explored. Here we report on a transgenic mouse line, named Tg-APP (Sw, V717F)/B6, that expresses the human amyloid precursor protein (APP) containing the Swedish and the V717F Indiana mutations in the brains of inbred C57BL/6 mice, designed to eliminate the potential phenotypic variations attributed to the compound genetic backgrounds adopted in most AD mouse models. The Tg-APP (Sw, V717F)/B6 mice expressed the transgene transcript, in the heterozygote state, at a level of 2.6 +/- 0.1 fold higher than that of endogenous mouse APP. However, no Abeta-plaque deposition was produced in the brain of the Tg-APP (Sw, V717F)/B6 mice up to 18 months of age. The Tg-APP(Sw, V717F)/B6 mice at 13-15 months showed reduced expression of calbindin and c-Fos in the brain. The Tg-APP (Sw, V717F)/B6 mice at 11-14 months displayed decreased motor coordination, learning and memory deficits, and severely increased anxiety. These phenotypes were not observed in the Tg-APP (Sw, V717F)/B6 mice at 5-7 months. Microarray analysis revealed altered expression, in the amygdala of the Tg-APP (Sw, V717F)/B6 mice, of genes previously implicated in anxiety. Taken together, these results suggest that the transgenic APP, or its derivatives, produces the age-dependent pathophysiology of the AD-like brain and that the progressive cognitive impairment and anxiety induction can proceed in the absence of visible Abeta-plaque deposition. PMID- 15114631 TI - Epidemiological studies of radio frequency exposures and human cancer. PMID- 15114630 TI - GAD67 and GAD65 mRNA and protein expression in cerebrocortical regions of elderly patients with schizophrenia. AB - Gamma-Aminobutyric acid (GABA), the principal inhibitory neurotransmitter of CNS, has been consistently implicated in the pathophysiology of schizophrenia. GABA is synthesized from glutamate by the enzyme glutamic acid decarboxylase (GAD). Two isoforms of GAD have been identified and have been named GAD65 and GAD67 based on their apparent molecular weights. In this study, GAD65 and GAD67 mRNA and protein levels were measured by using real-time RT-PCR and immunoblotting, respectively, in post-mortem brain tissue from the dorsolateral prefrontal cortex (DLPFC) and the occipital cortex of the elderly persons with schizophrenia and matched normal controls. In addition, the mRNA expression of GAT-1, one of the principal transporters of GABA, was also studied in the same subjects. Expression of GAD65 and GAD67 mRNA in the DLPFC and in the occipital cortex was significantly elevated in patients with schizophrenia, whereas the expression of the corresponding proteins and GAT-1 mRNA was unchanged. Although the levels of GAD65 and GAD67 messages were increased in schizophrenia subjects, the proportion of the two GAD isoforms remained constant in controls and schizophrenics. In the human DLPFC, GAD65 mRNA was found to be expressed significantly less than the message for GAD67, approximately 16% of that observed for GAD67. On the contrary, the abundance of GAD65 protein in the DLPFC was about 350% of that observed for GAD67. The results suggest a substantial dysregulation of GAD mRNA expression in schizophrenia and, taken together with the results of protein expression studies, raise the possibility that both cortical and subcortical GABA function may be compromised in the disease. PMID- 15114632 TI - Lack of adverse effects in pregnant/lactating female rats and their offspring following pre- and postnatal exposure to ELF magnetic fields. AB - We have recently reported that exposure of pregnant rats to 60 Hz at field strengths up to 0.5 mT during the entire period of pregnancy did not induce any biologically significant effects on both pregnant dams and embryo-fetal development. The present study was carried out to investigate the potential effects of gestational and lactational MF exposure on pregnancy, delivery, and lactation of dams and growth, behavior, and mating performance of their offspring in rats. Timed-pregnant female Sprague-Dawley (SD) rats (24/group) received continuous exposure to 60 Hz magnetic field (MF) at field strengths of 0 (sham control), 5 microT, 83.3 microT, or 0.5 mT. Dams received MF or sham exposures for 21 h/day from gestational day 6 through lactational day 21. Experimentally generated MF was monitored continuously throughout the study. No exposure-related changes in clinical signs, body weight, food consumption, pregnancy length, and necropsy findings were observed in dams. Parameters of growth, behavior, and reproductive performance of offspring showed no changes related to MF exposure. There were no adverse effects on embryo-fetal development of F2 offspring from dams exposed to MF. In conclusion, exposure of pregnant SD rats to 60 Hz at field strengths up to 0.5 mT from gestational day 6 to lactational day 21 did not produce biologically significant effects in dams, F1 offspring, or F2 fetuses. PMID- 15114633 TI - Extremely low frequency magnetic field effects on premorbid behaviors produced by cocaine in the mouse. AB - We investigated the premorbid behavioral changes produced by the administration of cocaine and acute exposure to extremely low frequency (ELF) magnetic field (MF) in the mouse. ICR mice received intraperitoneal injections of cocaine at two doses (65 and 70 mg/kg) and were subsequently exposed to one of eight ELF-MF fields (2, 3, 4, 8, 10, 15, 25, or 60 Hz) of about 20 G (2 mT) intensity immediately after injection. Twelve mice were used for each of applied cocaine dose and ELF-MF level. For a given dose of cocaine, the applied MF frequencies were randomly ordered, and blind tests were carried out in which the behavior observer did not know the frequencies of MF. The premorbid behaviors were defined in the ICR mice and their changes were observed over the exposure of various ELF MFs. Our data show that the onset times of stop rearing and tonic-clonic seizure in the 4 Hz MF exposure group are significantly different from those of the sham group. PMID- 15114634 TI - An analytical model for the calculation of the change in transmembrane potential produced by an ultrawideband electromagnetic pulse. AB - The electric field pulse shape and change in transmembrane potential produced at various points within a sphere by an intense, ultrawideband pulse are calculated in a four stage, analytical procedure. Spheres of two sizes are used to represent the head of a human and the head of a rat. In the first stage, the pulse is decomposed into its Fourier components. In the second stage, Mie scattering analysis (MSA) is performed for a particular point in the sphere on each of the Fourier components, and the resulting electric field pulse shape is obtained for that point. In the third stage, the long wavelength approximation (LWA) is used to obtain the change in transmembrane potential in a cell at that point. In the final stage, an energy analysis is performed. These calculations are performed at 45 points within each sphere. Large electric fields and transmembrane potential changes on the order of a millivolt are produced within the brain, but on a time scale on the order of nanoseconds. The pulse shape within the brain differs considerably from that of the incident pulse. Comparison of the results for spheres of different sizes indicates that scaling of such pulses across species is complicated. PMID- 15114635 TI - Use of a permanent magnetic field to inhibit the development of canine osteoarthritis. AB - This study was designed to determine the potential of a permanent magnetic field to inhibit the progression of osteoarthritis (OA) in a canine model. The magnetic field was created by 72 domino-sized ceramic magnets with surface field strength of 1100 G (0.11 T). The magnetic field strength at the surface of the mattress was 450-500 G (45-50 mT) and was equally distributed over the mattress surface. Eighteen animals had closed resection of their right stifle anterior cruciate ligament. Their kennel floors were covered in one of three ways: no floor mattress (OA) (N = 6); a floor mattress with domino-sized ceramic pieces placed between two layers of foam (sham control OA-MAT) (N = 6); or a floor mattress with domino-sized ceramic permanent magnets placed between two layers of foam (OA MAT-MAG) (N = 6). Animals were kept in their cages except for 4 h of exercise each day. The left stifle of six animals served as the normal control. The stifle joints were examined at 12 weeks for synovial effusion, gross anatomic appearance, microscopic anatomic appearance (Mankin score), and metalloproteinase (MMP)-1 and -3. Macroscopically, the OA-MAT-MAG group appeared to have less synovitis, less synovial effusion, less disruption of the cartilage surface, and less cartilage ulceration than did the OA group or the control mattress group. The mean Mankin score for the OA-MAT-MAG group was less than that for the OA group (4.2 +/- 0.8 vs. 6.7 +/- 0.3; P <.05) and the control mattress group (4.2 +/- 0.8 vs. 5.2 +/- 0.8; P >.05), but greater than that for the normal left group (4.2 +/- 0.8 vs. 1.0 +/- 0.4; P <.05). These scores show a trend of improvement for OA-MAT-MAG group but the difference with the sham control OA-MAT group was not statistically significant. In immunohistochemical studies, the OA-MAT-MAG group cartilage was stained less heavily for MMP-1 and MMP-3 than were the OA group cartilage and the control mattress group cartilage, but did not differ significantly in MMP-1 and MMP-3 from the normal left group cartilage. The OA-MAT MAG group did not differ from the normal left group in MMP-3 as determined by Western blot analysis. The study suggests that OA of the medial femoral condyle developed in a canine model exposed to a magnetic field may be inhibited beyond the benefit provided by mattress. Further studies are needed to delineate more precisely the effect of the magnetic field in reducing the severity of OA. PMID- 15114636 TI - The effect of short, high intensity magnetic field pulses on the healing of skin wounds in rats. AB - The object of this study was to examine the effect of high intensity, short duration pulsed electromagnetic fields (PEMF) on the healing of full thickness skin wounds in rats. Full thickness skin wounds were surgically created in two groups of Sprague-Dawley male rats. The rats were randomly divided into two groups, each containing 20 rats. Animals in the treatment group received treatments with the PEMF device on day 0, 3, 7, 9, 12, 14, 17, and 22, while the rats in the control group were subjected to the same procedure, but with the PEMF device not activated. Photographs of the surgically created wounds were obtained on day 0, 3, 7, 9, 12, 14, 17, and 22. Wound contraction (WC), wound epithelialization (WE), non-healed wound, and contraction-epithelialization (CE) ratio were calculated for each wound. No significant difference was found between the two groups for the parameters of WC, WE, non-healed wound, and CE ratio. A significant group x time interaction was found for WE and CE ratio. This type of PEMF did not have a significantly beneficial effect on wound healing. Wounds in the PEMF treated group were relatively less contracted and showed a compensatory increase in epithelialization in the early stages of wound repair. PMID- 15114637 TI - Magnetic field from spot welding equipment--is the basic restriction exceeded? AB - A point measurement of the magnetic field (MF) near a typical spot welding machine showed that the magnetic flux density was above the ICNIRP reference level. To investigate if the basic restrictions were exceeded, the induced body currents in a full 3D human model were calculated. It was found that at an operator position of 34 cm away from the machine the maximum induced current density was below the ICNIRP basic restriction. But if the operator was closer to the machine, the basic restriction was exceeded. An important finding of the present article is that the basic restriction can be exceeded although the MF spatially averaged over the whole body is well below the reference level. PMID- 15114638 TI - Metallic electrodes and leads in simultaneous EEG-MRI: specific absorption rate (SAR) simulation studies. AB - The purpose of this study was to investigate the changes in specific absorption rate (SAR) in human-head tissues while using nonmagnetic metallic electroencephalography (EEG) electrodes and leads during magnetic resonance imaging (MRI). A realistic, high resolution (1 mm(3)) head model from individual MRI data was adopted to describe accurately thin tissues, such as bone marrow and skin. The RF power dissipated in the human head was evaluated using the FDTD algorithm. Both surface and bird cage coils were used. The following numbers of EEG electrodes/leads were considered: 16, 31, 62, and 124. Simulations were performed at 128 and 300 MHz. The difference in SAR between the electrodes/leads and no-electrodes conditions was greater with the bird cage coil than with the surface coil. The peak 1 g averaged SAR values were highest at 124 electrodes, increasing to as much as two orders of magnitude (x172.3) at 300 MHz compared to the original value. At 300 MHz, there was a fourfold (x3.6) increase of SAR averaged over the bone marrow, and a sevenfold (x7.4) increase in the skin. At 128 MHz, there was a fivefold (x5.6) increase of whole head SAR. Head models were obtained from two different subjects, with an inter-subject whole head SAR variability of 3%. . PMID- 15114639 TI - High frequency electromagnetic fields (GSM signals) affect gene expression levels in tumor suppressor p53-deficient embryonic stem cells. AB - Effects of electromagnetic fields (EMF) simulating exposure to the Global System for Mobile Communications (GSM) signals were studied using pluripotent embryonic stem (ES) cells in vitro. Wild-type ES cells and ES cells deficient for the tumor suppressor p53 were exposed to pulse modulated EMF at 1.71 GHz, lower end of the uplink band of GSM 1800, under standardized and controlled conditions, and transcripts of regulatory genes were analyzed during in vitro differentiation. Two dominant GSM modulation schemes (GSM-217 and GSM-Talk), which generate temporal changes between GSM-Basic (active during talking phases) and GSM-DTX (active during listening phases thus simulating a typical conversation), were applied to the cells at and below the basic safety limits for local exposures as defined for the general public by the International Commission on Nonionizing Radiation Protection (ICNIRP). GSM-217 EMF induced a significant upregulation of mRNA levels of the heat shock protein, hsp70 of p53-deficient ES cells differentiating in vitro, paralleled by a low and transient increase of c-jun, c myc, and p21 levels in p53-deficient, but not in wild-type cells. No responses were observed in either cell type after EMF exposure to GSM-Talk applied at similar slot-averaged specific absorption rates (SAR), but at lower time-averaged SAR values. Cardiac differentiation and cell cycle characteristics were not affected in embryonic stem and embryonic carcinoma cells after exposure to GSM 217 EMF signals. Our data indicate that the genetic background determines cellular responses to GSM modulated EMF. Bioelectromagnetics 25:296-307, 2004. PMID- 15114640 TI - Lack of effect of 10 kV/m 60 Hz electric field exposure on pregnant dairy heifer hormones. AB - Sixteen pregnant Holstein heifers weighing 521 +/- 46 kg, at 3.3 +/- 0.7 months of gestation and 2.2 +/- 2.0 months of age were confined to wooden metabolism cages and were exposed to a vertical electric field (EF) of 10.0 +/- 0.4 kV/m and an artificial light cycle of 12 h light-12 h dark. The heifers were divided into two replicates of eight each. Each replicate was divided into two groups of four animals each, one group becoming the non-exposed and the second, the EF exposed group. The exposed group were housed in metabolism cages in an area where EF were generated, and the non-exposed group, in metabolism cages located in the adjacent area where the EF was less than 2% of that present in the exposed area. The test animals were subject to the different treatments for 4 weeks continuously. After 4 weeks, the animals switched treatment, the exposed group becoming the non exposed group and vice-versa. Then the treatment continued for 4 more weeks. Catheters were inserted into the jugular vein of the animals, and blood samples were collected on twice a week to estimate the serum concentration of progesterone (P4), melatonin (MLT), prolactin (PRL), and insulin-like growth factor-1 (IGF-1). Feed consumption was measured daily and feed samples were collected twice a week. The results indicated that exposure of dairy cattle to EF similar to those encountered directly underneath a 735 kV high tension electrical power line carrying a maximum load of current, cannot be associated with any variation in the experimental variables mentioned above. An exception to this, is the variation in MLT, which was associated with the EF exposure. Due to the inconsistency of the MLT response in the different replicates, caution should be exercised in the interpretation of this phenomenon. PMID- 15114641 TI - Experimental evidence of a potentially increased thrombo-embolic disease risk by domestic electromagnetic field exposure. AB - We have used the EaHy926 endothelial cell line, able to secrete both pro and anti aggregant platelet agents, as a model for thrombo-embolic diseases. We experimentally established, by comparing these two secretions with or without a Faraday cage, that the environmental electromagnetic field significantly increases the thrombo-embolic risks in this endothelial cell line. PMID- 15114642 TI - Effect of static magnetic field on growth and sporulation of some plant pathogenic fungi. AB - Growth and sporulation of phytopathogenic microscopic fungi were studied under a static magnetic field. The applied flux densities were 0.1, 0.5, and 1 mT. The magnetic field decreased the growth of colonies by 10% using this flux density region. At 0.1 mT flux density, the deviations are significant, P =.001, while in other cases the deviations generally are not significant. At the same time, the number of the developed conidia of Alternaria alternata and Curvularia inaequalis increased by 68-133%, but the number of Fusarium oxysporum conidia decreased by 79-83%. The deviations are generally significant at the P =.05 level. PMID- 15114644 TI - Theophilus Painter: First steps toward an understanding of the human genome. PMID- 15114645 TI - Ultrastructural aspects of gonadal morphogenesis in Bufo bufo (Amphibia Anura) 1. Sex differentiation. AB - The morphogenesis of gonads in Bufo bufo tadpoles was studied, and ultrastructural differences between sexes were identified. All specimens analyzed initially developed gonads made up of a peripheral fertile layer (cortex) surrounding a small primary cavity. Subsequently a central layer of somatic cells (medulla) developed. Both layers were separated by two uninterrupted basal laminae between which a vestige of the primary cavity persisted. During female differentiation, the peripheral layer continued to be the fertile layer. In males, the central layer blended into the peripheral layer and the basal laminae disappeared. The somatic cells of the central layer came into direct contact with the germ cells; this did not occur in females. Testicular differentiation continued with the migration of germ cells towards the center of the gonad. The somatic elements surrounding the germ cells appeared to play an active role in their transfer to the center of the gonad. The peripheral layer shrank and became sterile. Two basal laminae then re-formed to separate the fertile central layer from the peripheral sterile one. Germ cells have always been thought to perform a passive role in sex differentiation in amphibians. Following the generally accepted "symmetric model", the mechanism of gonad development is symmetrical, with cortical somatic cells determining ovarian differentiation and medullary somatic cells determining testicular differentiation. In contrast, we found that sex differentiation follows an "asymmetric" pattern in which germ cells tend primarily toward a female differentiation and male differentiation depends on a secondary interaction between germ cells and medullary somatic cells. PMID- 15114646 TI - Stage-dependent modulation of limb regeneration by caffeic acid phenethyl ester (CAPE)--immunocytochemical evidence of a CAPE-evoked delay in mesenchyme formation and limb regeneration. AB - Caffeic acid phenethyl ester (CAPE), a natural compound of bee propolis, selectively inhibits proliferation of transformed cells in several cancer models in vitro. To examine in vivo CAPE function, we used the newt regeneration blastema as a model system wherein the processes of de-differentiation and subsequent proliferation of undifferentiated cells mimic changes associated with oncogenic transformation and tumorigenesis. We have shown that a single dose of CAPE significantly increased cell proliferation at the stages of blastema growth and re-differentiation. At the de-differentiation stage, CAPE significantly stimulated proliferation of wound epidermis keratinocytes, but decreased proliferation in the blastema mesenchyme. Immunohistochemistry with a mesenchymal cell marker, vimentin, revealed a highly significant reduction of vimentin staining in the mesenchyme of CAPE-treated regenerates (p<0.001). These results, together with morphological observations indicate that, at the de-differentiation stage, CAPE stimulated wound re-epithelization, increased keratinocyte proliferation and increased thickness of the wound epidermis. However, CAPE inhibited mesenchyme formation and proliferation. The functional consequence of the CAPE inhibitory action was a delay in limb regeneration. PMID- 15114647 TI - Prenatal and postnatal effects of corticosterone on behavior in juveniles of the common lizard, Lacerta vivipara. AB - Many animals exhibit dramatic responses when subjected to a stressor. A classic marker of the stress response is an increase in plasma glucocorticoids, but this constitutes only one step in the cascade from experience of a stressor to wider organismal changes, including behavior. The behavioral sensitivity to glucocorticoids would determine the consequences of the stress-related alteration of behavior for the organism. In this study we explored, under laboratory conditions, the prenatal and postnatal effects of corticosterone on activity and thermoregulation of juveniles of the common lizard, Lacerta vivipara. Activity was measured as the time spent moving and the time spent scratching the wall in an empty terrarium. Thermoregulatory behavior was measured as the time spent motionless under a light bulb. Activity and thermoregulation of juveniles of the common lizard showed a different sensitivity to prenatal and postnatal corticosterone treatment, modulated by juvenile sex and maternal condition. Prenatal corticosterone manipulation influenced the time spent moving in both sexes. By contrast, only juvenile females increased the time spent scratching the walls of the terrarium when corticosterone was delivered both at the prenatal and postnatal stage. Prenatal hormone manipulation increased the time spent basking by juveniles issued from large females. These results suggest that, in addition to influencing a variety of behavioral and morphological traits, corticosterone may also play an important role in the regulation of activity and thermoregulation of juvenile lizards, modulated by individual sex and maternal condition. PMID- 15114648 TI - Effect of testosterone on T cell-mediated immunity in two species of Mediterranean lacertid lizards. AB - One of the primary assumptions of the immunocompetence handicap hypothesis is that testosterone has an immunosuppressive effect, but conflicting results have been reported in a variety of bird species concerning the effect of testosterone on the humoral and the T cell-mediated components of the immune system. The T cell-mediated component of the immune system is particularly important during the breeding season, because the likelihood of injury during sexual competition is high and T cell-mediated immunity is essential for healing wounds and resisting infection. In this study we examined the effect of experimentally increased levels of testosterone during breeding season on T cell-mediated immunity in male lizards of two Mediterranean lacertid species, Psammodromus algirus and Acanthodactylus erythrurus. The hormonal treatment significantly increased testosterone of the experimental individuals. T cell-mediated responses to phytohemagglutinin stimulation were significantly suppressed in testosterone treated males of both species. Furthermore, there was a significant negative relationship between individual variability in T cell-mediated responsiveness and plasma testosterone concentration. The present study is the first to demonstrate testosterone-induced suppression of T cell-mediated immunity in lizards. PMID- 15114649 TI - Paracellular pathway in the shell epithelium of Anodonta cygnea. AB - Ultrastructural study of cell-cell connections in the outer mantle epithelium (OME) on high-pressure-frozen specimens revealed zonula adherens, septate junctions and gap junctions in Anodonta cygnea. In order to evaluate the permeability of the paracellular pathway, the OME was incubated under gradients of lanthanum and calcium. After lanthanum incubation (4 mM) from the basal side, the septate junctions were penetrated completely by this tracer. When applied from the apical side, lanthanum deposits were located similarly over the entire length of the septate junctions up to the first dilatations of the intercellular space. Calcium deposits were also present in paracellular areas only when OME had been incubated simultaneously with calcium (6 mM) and lanthanum (4 mM) gradients. Lanthanum and calcium deposits were detected with ESI (Electron Spectroscopic Imaging) and identified with EELS (Electron Energy Loss Spectroscopy). On the other hand, electrophysiological observations showed a 48% reduction of conductance when the OME was bathed on both sides with solutions containing lanthanum (4 mM) and calcium (6 mM), compared to bathing with lanthanum-free solution (control). The conductance reduction was 52% when calcium was removed from the control solution. Supported by morphological and physiological evidence, it appears that, under in vivo conditions, calcium ions may diffuse paracellularly from the haemolymph towards the extrapallial fluid and vice-versa across the septate junctions in the OME of A. cygnea. Permeability of the septate junctions depended proportionally on the calcium concentration in fluids. PMID- 15114650 TI - Sexually dimorphic basal water absorption at the isolated pelvic patch of Japanese tree frog, Hyla japonica. AB - Frogs ingest little water orally, but absorb the majority of the water needed for normal physiological performance through a specific region of the ventral skin, the pelvic patch. We observed non-stimulated (basal water absorption, BWA) water flux through the isolated pelvic patch in vitro in Japanese tree frog (Hyla japonica). We found that water flux through non-stimulated skin from the pelvic patch was greater in males than females. This water flux was confirmed as BWA by observing no effect following the in vitro administration of propranolol and [adamantaneacetyl(1), O-Et-D-Tyr(2), Val(1), aminobutyryl(6), Arg(8, 9)] vasopressin, which are a beta-adrenergic receptor antagonist and a vasopressin V2 receptor antagonist, respectively. We further examined this phenomenon following gonadectomy, treatment with sex hormones (E2, 17beta-estradiol; TP, testosterone propionate), estrogenic chemicals (BPA, bisphenol A; MTX, methoxychlor) or prolactin (PRL, a hormone regulated by sex hormones that has osmoregulatory activity). Ovariectomy increased BWA in females. Injection (in vivo treatment) of E2 or PRL reduced BWA in males, whereas TP injection increased BWA in females. However, the in vitro addition of E2, TP, or PRL to the Ringer's solution on the serosal side of the ventral skin patch did not alter BWA. Subsequently, we injected (in vivo treatment) BPA or MTX, environmental chemical contaminants with known hormonal actions in mammals. Injection of BPA or MTX reduced BWA in males as observed following treatment with E2. These results provide the first evidence of sexual dimorphism in BWA through the isolated pelvic patch. The gonad appears essential for observed sexual dimorphism in BWA, and we hypothesize that sex hormones regulate the release of PRL, that in turn influences BWA indirectly. E2 is known to exert a specific stimulatory effect on PRL secretion. In addition, we have observed that some endocrine disrupting contaminants also eliminate the sexual dimorphism in BWA observed in the Japanese tree frog. PMID- 15114651 TI - Potential role for the sorbitol pathway in the meiotic dysfunction exhibited by oocytes from diabetic mice. AB - Complications common to type I diabetes, such as cataracts and cardiovascular disorders, have been associated with activation of the polyol pathway, which converts glucose to fructose via the intermediate, sorbitol. Under normal glycemic conditions, glucose is typically targeted for glycolysis or the pentose phosphate pathway through phosphorylation by hexokinase. When glucose levels are elevated under diabetic conditions, hexokinase becomes saturated, and the excess glucose is then shunted to aldose reductase, which converts glucose to sorbitol. In the present study, we examined the potential effects of this pathway on the maturation process in mouse oocytes. Increasing concentrations of sorbitol suppressed FSH-induced maturation in oocytes from control mice. Culturing oocytes from diabetic mice in the presence of inhibitors of aldose reductase reversed the suppression of FSH-induced meiotic maturation. When oocytes from control mice were cultured with activators of aldose reductase, FSH-induced maturation was compromised. In addition, treatment with sorbitol or activators of the polyol pathway led to reduced cell-cell communication between the oocyte and the cumulus cells, as well as compromised FSH-mediated cAMP production and de novo purine synthesis. These data indicate that the suppression of FSH-induced meiotic maturation observed in oocytes from diabetic mice may result from a shunting of glucose through the polyol pathway. PMID- 15114652 TI - Structure and expression of Furin mRNA in the ovary of the medaka, Oryzias latipes. AB - A cDNA for furin was cloned from the ovary of the medaka, Oryzias latipes, by a combination of cDNA library screening, 5'-rapid amplification of cDNA ends (RACE), and 3'- RACE. The cDNA sequence codes for a protein of 814 amino acid residues highly homologous to other vertebrate furins, Ca(2+)-dependent serine proteases belonging to the subtilysin-like proprotein convertase family. The medaka preprofurin consists of a leader sequence, a propeptide with autoactivation sites, a Kex2-like catalytic domain, a P domain, a cysteine-rich domain, a putative transmembrane domain, and a cytoplasmic domain. The catalytic triad residues (Asp-164, His-205, and Ser-379) were all conserved. Furin mRNA was expressed in many tissues of this, including the ovary. In the ovary, the greatest expression of furin mRNA occurred in oocytes of small growing follicles, as demonstrated by Northern blotting, RT-PCR, and in situ hybridization analysis. Temporary and spatial expression patterns of the medaka fish furin were similar to those of stromelysin-3 and MT5-MMP during oocyte growth and postnatal development. PMID- 15114656 TI - The somatic generation of immune recognition. 1971. AB - Antibody specificity is determined by structural v-genes that code for the amino acid sequences of the variable regions of antibody polypeptide chains. The present hypothesis proposes that the germ-cells of an animal carry a set of v genes determining the combining sites of antibodies directed against a complete set of certain class of histocompatibility antigens of the species to which this animal belongs. The evolutionary development of this set of v-genes in phylogeny is traced back to the requirements for cell to cell recognition in all metazoa. The hypothesis leads to a distinction between two populations of antigen sensitive cells. One population consists of cells forming antibodies against foreign antigens; these lymphocytes have arisen as mutants in clones descending from lymphocytic stem cells which expressed v-genes belonging to the subset (subset S) coding for antibody against histocompatibility antigens that the individual happens to possess. The other population consists of allograft rejecting lymphocytes that express v-genes of the remaining subset (subset A) coding for antibody against histocompatibility antigens of the species that the individual does not possess. The primary lymphoid organs are viewed as mutant breeding organs. In these organs (e.g. in the thymus), the proliferation of lymphocytes expressing the v-genes of subset S and the subsequent suppression of the cells of these "forbidden" clones, leads to the selection of mutants cells expressing v-genes that have been modified by spontaneous random somatic mutation. This process generates self-tolerance as well as a diverse population of antigen-sensitive cells that reflects antibody diversity. The proliferation in the primary lymphoid organs of lymphocytes expressing v-genes of subset A generates the antigen-sensitive cell population that is responsible for allo aggression. The theory explains how a functional immune system can develop through a selection pressure exerted by self-antigens, starting during a period in early ontogeny that precedes clonal selection by foreign antigens. The hypothesis provides explanations for the variability of the N-terminal regions of antibody polypeptide chains, for the dominant genetic control of specific immune responsiveness by histocompatibility alleles, for the relative preponderance of antigen-sensitive cells directed against allogeneic histocompatibility antigens, for antibody-idiotypes, for allelic exclusion, for the precommitment of any given antigen-sensitive lymphocyte to form antibodies of only one molecular species and for the cellular dynamics in the primary lymphoid tissues. PMID- 15114657 TI - TCR-MHC/peptide interactions: kissing-cousins or a shotgun wedding? AB - The purpose of this Commentary is to put into modern-day perspective Jerne's hypothesis that antigen receptors encoded in the genome have been evolutionarily selected for their ability to react with major histocompatibility proteins and that the process of eliminating self reactivity is the catalyst for the generation of diversity of antigen receptors. In writing his hypothesis Jerne was trying to deal with the obsession of the immune system with the MHC, an obsession that was manifest in his days by the strong reactions of the immune system with allogeneic MHC proteins. However, Jerne's hypothesis also took on other issues that were not understood at the time--issues that included lymphocyte selection and tolerance, the generation of somatic diversity and the ability of the MHC to control responses to other antigens. In so doing, Jerne generated a hypothesis that accounted remarkably satisfactorily for what was known in 1971. Whilst the details of much of the hypothesis have since turned out to be incorrect, in his ideas Jerne did anticipate many of the most interesting and surprising findings of the subsequent 33 years. PMID- 15114658 TI - Myelin oligodendrocyte glycoprotein-35-55 peptide induces severe chronic experimental autoimmune encephalomyelitis in HLA-DR2-transgenic mice. AB - The use of HLA class II-transgenic (Tg) mice has facilitated identification of antigenic T cell epitopes that may contribute to inflammation in T cell-mediated diseases such as rheumatoid arthritis and multiple sclerosis (MS). In this study, we compared the encephalitogenic activity of three DR2-restricted myelin determinants [mouse (m) myelin oligodendrocyte glycoprotein (MOG)-35-55, human (h)MOG-35-55 and myelin basic protein (MBP)-87-99] in Tg mice expressing the MS associated DR2 allele, DRB1*1501. We found that mMOG-35-55 peptide was strongly immunogenic and induced moderately severe chronic experimental autoimmune encephalomyelitis (EAE) with white matter lesions after a single injection in Freund's complete adjuvant followed by pertussis toxin. hMOG-35-55 peptide,which differs from mMOG-35-55 peptide by a proline for serine substitution at position 42, was also immunogenic, but not encephalitogenic, and was only partially cross reactive with mMOG-35-55. In contrast, MBP-87-99, which can induce EAE in double Tg mice expressing both HLA-DR2 and a human MBP-specific TCR, was completely non encephalitogenic in HLA-DR2-Tg mice lacking the human TCR transgene. These findings demonstrate potent encephalitogenic activity of the mMOG-35-55 peptide in association with HLA-DR2, thus providing a strong rationale for further study of hMOG-35-55 peptide as a potential pathogenic determinant in humans. PMID- 15114659 TI - CD8 alpha is an activation marker for a subset of peripheral CD4 T cells. AB - Rat CD4 T lymphocytes express CD8 alpha upon activation. Here, we show that double-positive cells express CD8 alpha alpha homodimers, and we study their phenotype and function. Most activated CD4(+) lymphocytes expressing CD8 alpha are recent thymic emigrants. Accordingly, most activated CD4 single-positive thymocytes express CD8 alpha, and thymectomy and aging decrease the frequency of CD4(+)CD8 alpha(+) lymphocytes. However, CD8 induction is not restricted to CD4(+) recent thymic emigrants. CD4(+)CD8 alpha(+) and CD4(+)CD8 alpha(-)cells were generated in vitro from naive or from primed donors and, to study their function, were transferred to normal rats. Both cell types helped primary humoral responses, but only CD4(+)CD8 alpha(-) cells promoted secondary responses. Thus, memory CD4 T cells mediating antibody responses and some naive CD4(+) lymphocytes do not express CD8 alpha. In addition, CD4(+)CD8 alpha(+) cells produce mainly Th1 cytokines while CD4(+)CD8 alpha(-) cells produce IL-10 and showed a sustained proliferative response. Hence, CD8 alpha expression after activation distinguishes two distinct CD4 T cell subsets. PMID- 15114660 TI - Coupling of antigen to cholera toxin for dendritic cell vaccination promotes the induction of MHC class I-restricted cytotoxic T cells and the rejection of a cognate antigen-expressing model tumor. AB - We previously demonstrated that cholera toxin (CT) is highly efficient as a combined carrier and adjuvant for dendritic cell (DC) vaccination, inducing strong Th1-dominated B cell and CD4(+) T cell responses. In this study we show that vaccination with DC pre-pulsed ex vivo with CT-conjugated OVA (OVA-CT) gives rise to OVA-specific CD8(+) T cells that produce IFN-gamma and are cytotoxic for OVA-expressing E.G7 tumor cells both in vitro and in vivo. The induction of specific CD8(+) CTL by OVA-CT-treated DC was associated with enhanced presentation of OVA peptide (SIINFEKL) on MHC class I in combination with an overall activation of the pulsed DC. Vaccination of mice with OVA-CT-pulsed DC resulted in rejection of already established MHC class I-positive, MHC class II negative, OVA-expressing E.G7 tumors in an antigen-specific, CD8(+) T cell dependent fashion and was associated with high numbers of tumor-infiltrating CD8(+) T cells. Conjugation of antigen to CT facilitated DC uptake of the linked antigen through the GM1 receptor-binding B subunit and induced strong activation maturation signals through the biologically active A subunit. These results have interesting implications for DC vaccination aimed at inducing CTL immune responses. PMID- 15114661 TI - ICOS-mediated signaling regulates cytokine production by human T cells and provides a unique signal to selectively control the clonal expansion of Th2 helper cells. AB - The CD28 homologue inducible costimulator (ICOS) has been demonstrated to regulate a number of T cell-dependent immune responses in vivo. However, the expression and functional importance of ICOS during APC-Th cell interaction in the human is not fully understood. Here, we demonstrate that ICOS-mediated signaling plays an important role in the production of selective cytokines during both primary and subsequent Th cell responses upon allospecific or superantigen activation. In contrast, ICOS does not play a role in the differentiation of naive cells into Th1 or Th2 effector cells, nor does it determine the type of effector function of memory cells upon subsequent allogeneic challenge. In addition, our data demonstrate that ICOS provides a novel and unique role in regulating DC-mediated Th2, but not Th1 cell clonal expansion. These data suggest that ICOS-mediated signaling plays a discrete role in the regulation of human T helper cell responses. PMID- 15114662 TI - Granulocyte colony-stimulating factor promotes the generation of regulatory DC through induction of IL-10 and IFN-alpha. AB - We have recently demonstrated that G-CSF promotes the generation of human T regulatory (T(REG)) type 1 cells. In this study, we investigated whether the immunomodulatory effects of G-CSF might be mediated by DC. CD14(+) monocytes were cultured with serum collected after clinical administration of G-CSF (post-G), which contained high amounts of IL-10 and IFN-alpha. Similar to incompletely matured DC, monocytes nurtured with post-G serum acquired a DC-like morphology, expressed high levels of costimulatory molecules and HLA-DR, and exhibited diminished IL-12p70 release and poor allostimulatory capacity. Importantly, post G DC-like cells were insensitive to maturation stimuli. As shown by neutralization studies, IFN-alpha and, even more pronounced, IL-10 contained in post-G serum inhibited IL-12p70 release by post-G DC-like cells. Furthermore, phenotypic and functional features of post-G DC-like cells were replicated by culturing post-G monocytes with exogenous IL-10 and IFN-alpha. Post-G DC-like cells promoted Ag-specific hyporesponsiveness in naive allogeneic CD4(+) T cells and orchestrated a T(REG) response that was dependent on secreted TGF-beta 1 and IL-10. Finally, neutralization of IL-10 and IFN-alpha contained in post-G serum translated into abrogation of the regulatory features of post-G DC-like cells. This novel mechanism of immune regulation effected by G-CSF might be therapeutically exploited for tolerance induction in autoimmune disorders. PMID- 15114663 TI - Human CD25+ regulatory T cells: two subsets defined by the integrins alpha 4 beta 7 or alpha 4 beta 1 confer distinct suppressive properties upon CD4+ T helper cells. AB - Down-regulation of autoreactive T cell responses in vivo includes cell-contact dependent as well as contact-independent mechanisms. Infectious tolerance is a contact-dependent mechanism used by naturally occurring CD25(+) T regulatory cells (Tregs) to confer suppressive activity upon conventional CD4(+) T cells thereby generating secondary T helper suppressor cells(Th(sup)), which inhibit T cell activation via soluble mediators. Here, we describe two distinct subsets of human Tregs, characterized by expression of either the alpha(4)beta(7) integrin or the alpha(4)beta(1) integrin. Upon activation, both subsets show an enhanced expression of FoxP3, recently described as a key transcription factor of murine Tregs. In addition, both are able to convey suppressive capacity to conventional CD4(+) T cells. However, the properties of Treg subsets are rather distinct: alpha(4)beta(7) (+)Tregs induce IL-10-producing Th(sup) (Tr1-like), whereas alpha(4)beta(1) (+) Tregs induce TGF-beta-producing Th(sup) (Th3-like). Our findings reconcile conflicting results by clearly demonstrating that suppression through naturally occurring CD25(+) Tregs is primary cell-contact-dependent but is subsequently followed by cell-contact-independent T cell inhibition mediated by second-generation Tr1- and Th3-like Th(sup) via the soluble factors IL-10 and TGF-beta. PMID- 15114664 TI - Environmental pollutant tributyltin promotes Th2 polarization and exacerbates airway inflammation. AB - It has been shown that a relatively high dose of tributyltin (TBT), which is recognized as a particularly notable environmental pollutant, exerts immunotoxic effects such as thymic atrophy via induction of T cell apoptosis. However, the effect of low doses of TBT on the immune responses remains unknown. Here we show that environmentally relevant doses of TBT promoted strong Th2 polarization via suppression and augmentation of Th1 and Th2 development, respectively, from naive CD4(+) T cells primed with anti-CD3 and splenic antigen-presenting cells (APC). TBT-induced Th2 polarization was indirect, working through APC via suppression of IL-12 production by macrophages/DC and the augmentation of IL-10 production by B cells. Th2 polarization was also induced in mice treated with TBT and immunized with OVA or infected with Nippostrongylus brasiliensis. Furthermore, airway inflammation in mice sensitized and challenged with OVA was exacerbated by the administration of TBT with concomitant augmentation of Th2-type immunity. Our results highlight the fact that an important environmental pollutant TBT may present significant risk for the induction of allergic diseases via promotion of Th2 polarization. PMID- 15114665 TI - Impaired lymphocyte development and function in Clast5/Stra13/DEC1-transgenic mice. AB - Clast5/Stra13/DEC1 is a member of the helix-loop-helix family of transcriptional repressors. We have previously shown that Clast5 is rapidly down-regulated upon B cell activation and its overexpression inhibits cell cycle progression in B lymphoma cells. In the present study, we show that Clast5 expression is developmentally regulated during B cell differentiation, being expressed at the progenitor B cells, down-regulated at the precursor B cells, elevated in immature and mature resting B lymphocytes, and down-regulated again in germinal center B cells. To investigate the function of Clast5 in regulating lymphocyte development, we have generated transgenic mice expressing Clast5 in B- and T lineage cells (Clast5-Tg). Clast5-Tg mice grew and bred normally but their spleen and thymus cellularity was reduced compared with control littermates. The development of B cells in the bone marrow and T cells in the thymus was impaired, with the expansion of progenitor B and T cells most strongly affected. The frequency of IL-7-responsive cells in the bone marrow of Clast5-Tg mice was reduced by >80% and their proliferative response to IL-7 was also compromised. Mature B cells from Clast5-Tg mice were hyporesponsive to antigen receptor cross linking and exhibited mild reduction in the proliferative response to CD40 ligation or lipopolysaccharide stimulation. Moreover, the development of germinal center B cells and antibody production against a T-dependent antigen were reduced in Clast5-Tg mice. These results reveal a critical role for Clast5/Stra13/DEC1 in negatively regulating lymphocyte development and function in vivo. PMID- 15114666 TI - Differential usage of VLA-4 and CXCR4 by CD3+CD56+ NKT cells and CD56+CD16+ NK cells regulates their interaction with endothelial cells. AB - The mechanism that regulates the preferential accumulation of NKT cells in the BM is unknown. The BM endothelium constitutively expresses selectins, the integrin ligands VCAM-1 and ICAM-1, and the chemokine CXCL12. Both NK and NKT subsets of cells exhibited similar tethering and rolling interactions on both P-selectin and E-selectin and expressed similar levels of the integrins, VLA-4 and LFA-1. Although NKT cells express higher levels of CXCR4 than NK cells, CXCL12 (the ligand for CXCR4) rapidly stimulates similar levels of adhesion of NK and NKT cells to VCAM-1 and ICAM-1. In both subsets, the arrest on VCAM-1 was dependent on high affinity VLA-4 and the homing of these cells to the BM of NOD/SCID was VLA-4-dependent. However, as opposed to the situation for NK cells, CXCL12 preferentially triggers, under shear flow, the rolling on VCAM-1 and transendothelial migration of NKT cells. Moreover, over-expression of high levels of CXCR4 on the YT NK cell line enables them to migrate in response to CXCL12. This study therefore suggests an important role for CXCR4 levels of expression and for VLA-4 in regulating the accumulation of NKT cells in the BM. PMID- 15114667 TI - CD38 is expressed on human mature monocyte-derived dendritic cells and is functionally involved in CD83 expression and IL-12 induction. AB - Dendritic cell (DC) maturation is characterized by the gain or loss of immunological functions and by expression of distinctive surface receptors. CD38 is an ectoenzyme that catalyzes the synthesis of cyclic ADP ribose (a potent second messenger for Ca(2+) release), as well as a receptor that initiates transmembrane signaling upon engagement with its counter-receptor CD31 or with agonistic monoclonal antibodies. Since CD38 is expressed by resting monocytes, we aimed to monitor CD38 expression during the differentiation of human monocyte derived DC (MDDC) and to investigate the possibility that CD38 plays a functional role during DC maturation. CD38 is down-modulated during differentiation into immature MDDC and expressed again upon maturation. The extent of CD38 expression is dependent on the stimulus adopted (LPS > IFN-gamma > CD40 cross-linking). Although weak, IFN-gamma consistently induces DC maturation. De novo-synthesized CD38 is enzymatically active, and its expression in mature (m) MDDC is dependent on NF-kappa B activity. However, CD38 is not merely a maturation marker but also mediates signaling in mMDDC, where it maintains its functions as a receptor. Activation via agonistic anti-CD38 mAb induces up-regulation of CD83 expression and IL-12 secretion, whereas disruption of CD38/CD31 interaction inhibits CD83 expression, IL-12 secretion and MDDC-induced allogeneic T cell proliferation. PMID- 15114668 TI - Transient gain of effector function by CD8+ T cells undergoing peripheral tolerance to high-dose self-antigen. AB - Induction of peripheral T cell tolerance is mediated by bone marrow-derived dendritic cells that cross-present self-antigen to self-reactive T cells. The current model for peripheral CD8(+) T cell tolerance is that TCR engagement by self-antigen in the absence of costimulation results in abortive activation without development of effector function. Here we demonstrate in vivo that high dose self-antigen ("signal 1") can compensate for lack of costimulation ("signal 2"), leading to full activation of and development of effector function by self reactive T cells. In the setting of low-dose self-antigen, acquisition of effector function by self-reactive T cells is dependent on costimulation via CD40 ligation in vivo. However, gain of effector function in either setting does not prevent eventual tolerance of self-reactive CD8(+) T cells. These results suggest that the mechanisms for peripheral CD8(+) T cell tolerance are more complex than the proposed "signal 1 in the absence of signal 2" hypothesis. Further exploration of these mechanisms will have direct impact on the design of effective immunotherapy for autoimmune diseases, chronic infections and cancers. PMID- 15114669 TI - Chronic graft-versus-host reaction is associated with a decrease in Ig light chain receptor editing in bone marrow self-reactive B cells. AB - The encounter of developing B cells in the bone marrow with soluble hen egg lysozyme (sHEL) self antigen induces anergy and endogenous kappa light chain rearrangements ('receptor editing'). We have previously shown that induction of chronic graft-versus-host reaction (GVH) in tolerant Ig/sHEL mice results in prevention of B cell anergy in the bone marrow and the spleen. We now report that in chronic GVH, immature self-reactive B cells also show reduced levels of receptor editing in the bone marrow. This is evidenced by the following observations: (a) a small population of'receptor-edited' B cells, which is found in tolerant mice, is markedly reduced in mice that have lost tolerance in chronic GVH; (b) self-reactive B cells in GVH mice have reduced levels of endogenous kappa chain rearrangements; and (c) recombinase-activating gene (RAG)-2 expression is markedly decreased in immature self-reactive B cells in the bone marrow of chronic GVH mice. These results suggest that in chronic GVH newly emerging B cells escape tolerance, in part because of decreased receptor editing in the bone marrow. Thus, the autoimmunity induced by chronic GVH may ultimately result from the failure of B cell tolerance at multiple checkpoints. PMID- 15114670 TI - Commensal Gram-negative bacteria prime human dendritic cells for enhanced IL-23 and IL-27 expression and enhanced Th1 development. AB - Dendritic cells (DC) are the main orchestrators of specific immune responses. Depending on microbial information they encounter in peripheral tissues, they promote the development of Th1, Th2 or unpolarized Th cell responses. In this study we have investigated the immunomodulatory effect of non-pathogenic intestinal Gram-negative (Escherichia coli, Bacteroides vulgatus,Veillonella parvula, Pseudomonas aeruginosa) and Gram-positive (Bifidobacterium adolescentis, Enterococcus faecalis, Lactobacillus plantarum and Staphylococcus aureus) bacteria on human monocyte-derived DC (moDC). None of the Gram-positive bacteria (GpB) primed for Th1 or Th2 development. In contrast, despite the low levels of IL-12 they induce, all Gram-negative bacteria (GnB) primed moDC for enhanced Th1 cell development, which was dependent on IL-12 and an additional unidentified cofactor. Strikingly, GnB-matured moDC expressed elevated levels of p19 and p28 mRNA, the critical subunits of IL-23 and IL-27, respectively, suggesting that the IL-12 family members may jointly be responsible for their Th1-driving capacity. Purified major cell wall components of either GnB or GpB did not yield Th cell profiles identical to those obtained with whole bacteria, and could not explain the induction of the IL-12 family members nor Th1 priming by GnB. Importantly, this study gives indications that the expression of the different IL-12 family members is dictated by different priming conditions of immature DC. PMID- 15114671 TI - In vivo activation of invariant V alpha 14 natural killer T cells by alpha galactosylceramide sequentially induces Fas-dependent and -independent cytotoxicity. AB - The present study was designed to clarify the cytotoxic capacities of invariant V alpha 14 natural killer T (iNKT) cells activated in vivo. We found that as early as 2 h after a single injection of alpha-galactosylceramide (alpha-GalCer), sorted iNKT splenocytes from treated mice kill Fas-transfected target cells. The implication of the Fas pathway in this lysis was strengthened by both the blockage of cytotoxicity in the presence of anti-Fas ligand (FasL) monoclonal antibody (mAb) and the up-regulation of FasL expression on iNKT cells. Sorted NK cells did not participate in the lytic activity at this time point. Yet, they became cytotoxic later on, 24 h post-treatment, when target cell lysis was mainly independent of the Fas pathway. This type of cell killing was predominant at this later time point, even though iNKT cells conserved a slight Fas-dependent cytotoxicity. NK cells failed to acquire the ability to kill target cells when IFN-gamma production in alpha-GalCer-injected mice was blocked by anti-IFN-gamma mAb, underscoring the major role of this cytokine. In conclusion, our findings provide the first direct evidence that iNKT cells can exert Fas-dependent cytotoxicity very shortly after in vivo alpha-GalCer activation and later, through IFN-gamma secretion, enable NK cells to kill target cells in a Fas independent pathway. PMID- 15114672 TI - Perforin and IFN-gamma do not significantly regulate the virus-specific CD8+ T cell response in the absence of antiviral effector activity. AB - Using gene-targeted mice we have investigated whether perforin and/or interferon gamma exert a direct regulatory effect on the expansion and contraction of antigen-specific CD8(+) T cells following infection with a virus (vesicular stomatitis virus) which is not controlled through these molecular effector systems. Unlike what has been observed when these molecules are essential for pathogen clearance, neither molecule was found to play an important role in regulating the kinetics of the virus-specific CD8(+) T cell response in the absence of antiviral effector activity. PMID- 15114673 TI - Increased NF-kappa B activity in B cells and bone marrow-derived dendritic cells from NOD mice. AB - Type 1 diabetes results from the breakdown of peripheral tolerance. As regulators of T cell activation, antigen-presenting cells (APC) modulate peripheral tolerance and hence contribute to the immune dysregulation characteristic of insulin-dependent diabetes mellitus (IDDM). We initially observed an increased importance of NOD B cell APC function in a T cell priming assay as compared to non-autoimmune strains. Consistent with this increased APC function, we found that NF-kappa B nuclear translocation is increased in unmanipulated NOD and NOD.B10Sn-H2(b) B cells and that, in addition, NOD B cells are more sensitive to NF-kappa B-activating stimuli. We obtained similar results using NOD bone marrow derived dendritic cell (BMDC) cultures. As costimulatory molecules have been shown to be NF-kappa B responsive, we examined the expression of these markers on NOD APC. Both B cells and BMDC expressed elevated levels of CD80 and CD40. Finally, NOD B cells provided better allostimulation than B cells from non autoimmune strains. Therefore, hyperactivation of NF-kappa B and increased expression of CD80 and CD40 by NOD B cells and BMDC may be a contributing factor in the selection of effector T cells observed in IDDM. PMID- 15114674 TI - Inhibition of Notch signaling biases rat thymocyte development towards the NK cell lineage. AB - Notch receptors are involved in directing the choice between alternative cell fates in developmental scenarios such as thymopoiesis. By pharmacological interference in rat fetal thymus organ culture we show that inhibition of Notch signaling arrests T cell development at an early double-negative stage and is accompanied by a dramatic increase in the number of NK cells. These cells show an activated phenotype, lack recombination of the TCR beta gene locus and express perforin. Similarly, in thymic lobes reconstituted with fetal liver cells, progenitors predominantly develop into NK cells both after pharmacological interference of Notch and after treatment with a recombinant rat Notch1/Fc chimera. Collectively, this identifies the lineage decision of NK/T precursor cells as an important site of Notch action in rat thymocytes. PMID- 15114675 TI - Expression of milk fat globule epidermal growth factor 8 in immature dendritic cells for engulfment of apoptotic cells. AB - Milk fat globule epidermal growth factor 8 (MFG-E8) is a protein that stimulates the engulfment of apoptotic cells by phagocytes. Here, we show that mouse immature dendritic cells (DC) generated in vitro by culturing bone marrow progenitors in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF), and Langerhans cells present in the skins, expressed MFG-E8. Bone marrow-derived macrophages generated by M-CSF did not express MFG-E8. MFG-E8 expressed in immature DC was found to be secreted as exosomes. The expression of MFG-E8 was significantly suppressed when the immature DC were induced to mature by treating them with lipopolysaccharides. This expression of MFG-E8 was well correlated with the ability of the cells to engulf apoptotic cells. That is,immature DC phagocytosed apoptotic cells more efficiently than did mature DC or bone marrow-derived macrophages. The ability of immature DC to engulf apoptotic cells was severely reduced when the immature DC were prepared from MFG E8-deficient mice. These results indicated that MFG-E8 plays an essential role in the engulfment of apoptotic cells by bone marrow-derived immature DC. PMID- 15114676 TI - Somatic mutations can lead to a loss of superantigenic and polyreactive binding. AB - Although antibodies have been assumed to bind a specific antigen, evidence exists showing that a single antibody can bind to multiple unrelated antigens. We previously studied a human monoclonal antibody expressing a mutated form of the V(H)3-73 gene and displaying anti-tubulin activity in a patient suffering from an immunocytic lymphoma. Despite its expression of a V(H)3 family member, this immunoglobulin failed to react with protein A (SpA), suggesting that somatic mutations could account for its change in specificity. To examine this possibility, we produced recombinant Ig expressing germ-line (IgM kappa-Germ) or the mutated form (IgM kappa-PER) of the V(H)3-73 fragment. Comparison of the respective affinities of the two Ig demonstrated that IgM kappa-Germ restores its SpA-binding capacity, and shows a moderate decrease in its affinity for tubulin. Interestingly, IgM kappa-Germ displayed polyreactive specificity for different autoantigens, which contrasted to the monospecific binding of IgM kappa-PER to tubulin. These results suggest that the monoreactive IgM kappa-PER antibody may be derived from a natural polyreactive antibody through somatic mutation. In addition, both temperature modification and mild denaturation succeeded in recovering the polyreactivity of IgM kappa-PER, which favors the view that conformational modifications of the tertiary structure of antibodies may play a key role in the genesis of polyreactivity. PMID- 15114677 TI - Immunotherapy with OX40L-Fc or anti-CTLA-4 enhances local tissue responses and killing of Leishmania donovani. AB - Enhancing granuloma development and effector function, but without inducing the pathology associated with excess granulomatous inflammation, poses a major challenge for immunotherapeutic intervention against diseases such as visceral leishmaniasis (VL). Here, we demonstrate that a chimeric fusion protein (OX40L Fc) which stimulates T cells through OX40 and a monoclonal antibody which blocks CTLA-4, an inhibitory receptor on T cells, both enhanced the rate of granuloma maturation, CD4(+) T cell proliferation, and killing of Leishmania. Costimulation based therapy induced no adverse fibrotic or necrotic reactions, and had no significant effect on the levels of endogenous anti-inflammatory cytokines (IL-10 and TGF-beta). Furthermore, both OX40L-Fc and anti-CTLA4 could be co-administered with conventional anti-leishmanial drugs. Until now, enhancing T cell immunity by the manipulation of costimulatory pathways has only received serious attention for cancer immunotherapy, but our data provide a compelling argument for the evaluation of this approach in human VL and other infectious diseases. PMID- 15114678 TI - Inhibition of hepatic transcriptional induction of lipopolysaccharide-binding protein by transforming-growth-factor beta 1. AB - LPS-binding protein (LBP) is an acute-phase protein with the ability to bind and transfer LPS of Gram-negative bacteria, as well as cell wall compounds of other pathogenic bacteria. This soluble pattern-recognition molecule is present in high concentrations in serum and represents an important defense mechanism of the host. Regulation of the hepatic acute-phase response and its termination are important mechanisms for limiting systemic inflammatory activity of the host organism. We show here that TGF-beta 1, in a dose-dependent fashion, is able to inhibit LBP transcript accumulation and LBP protein synthesis induced by IL-6, IL 1 beta and dexamethasone in hepatoma cell lines. These data were confirmed employing primary human hepatocytes, where TGF-beta 1 also inhibited LBP protein synthesis. We identified and analyzed several Smad-binding sites (Smads are major regulatory elements of TGF-beta 1) within the LBP promoter, and found that one of them was active. We furthermore identified an AP-1-binding site clearly conferring inhibitory effects of TGF-beta 1 towards LBP promoter activity, shown by gel shift and promoter mutagenesis experiments. Further elucidating the mechanism of transcriptional regulation of proteins involved in innate immune responses may potentially help to develop novel intervention strategies for the acute-phase response, sepsis, and septic shock. PMID- 15114679 TI - Porphyromonas gingivalis lipopolysaccharides induce maturation of dendritic cells with CD14+CD16+ phenotype. AB - Primary immune responses are initiated by dendritic cells (DC) that inform naive T helper cells about invading pathogens. DC undergo sequential events leading to irreversible maturation upon bacterial stimulation. To investigate the responses of DC during periodontal infection, we studied the effects of LPS from Porphyromonas gingivalis on DC. DC generated from human peripheral monocytes by culture with IL-4 and GM-CSF were incubated with P. gingivalis LPS (Pg LPS) or Escherichia coli LPS (Ec LPS). Flow cytometry and real-time quantitative RT-PCR analysis revealed that Pg LPS, but not Ec LPS, preferentially up-regulated CD14 and CD16 expression at protein and mRNA levels. Furthermore, Pg LPS preferentially induced the secretion of soluble CD14. CD1a, HLA-DR and CD54 were highly expressed on DC stimulated with both kinds of LPS; however, CD40, CD80, CD83 and CD86 expression on Pg LPS-stimulated DC was lower than on Ec LPS stimulated DC. With regard to IL-6, IL-8, IL-10, IL-12 and RANTES production from DC and allogeneic T cell proliferation, Pg LPS was a weaker stimulator than Ec LPS. These results suggested that Pg LPS triggers maturation of DC with unique characteristics, which exhibited weak immunostimulatory activity and may contribute to induction of chronic inflammation at the site of periodontal infection. PMID- 15114680 TI - Necrosis-like cell death induced by bacteria in mouse macrophages. AB - The death of individual cells is a frequent and physiological event in the mammalian immune system and most often occurs by apoptosis. It is becoming increasingly clear that cell death is also induced during bacterial infections. Here we report that, in addition to the apoptotic form already established, a necrosis-like form of cell death is induced by pyogenic bacteria (Enterobacteriaceae, Pseudomonas, enterococci) in mouse macrophages. Necrosis could be separated from apoptosis as it did not require phagocytosis of bacteria and occurred when apoptosis was inhibited by caspase blockade or by Bcl-2. Furthermore, ligands that stimulate Toll-like receptors were also found to have the capacity to induce necrosis. Strikingly, this form of cell death was sufficient for the uptake of dead cells by either mouse bone marrow-derived DC or a cell line derived from DC, possibly by virtue of the externalization of phosphatidylserine. Since the loading with bacteria-carrying cells is likely to impact on DC function, this form of necrosis may have a previously unsuspected role in the development of an immune response. PMID- 15114681 TI - Expression of L-histidine decarboxylase in granules of elicited mouse polymorphonuclear leukocytes. AB - Infiltrating polymorphonuclear leukocytes (PMN) in the peritoneal cavity were found to express L-histidine decarboxylase (HDC), the rate-limiting enzyme of histamine synthesis, in a csein-induced peritonitis model. Expression of HDC was detected in the elicited PMN, but not in the peripheral blood leukocytes. The peritoneal lavage fluids in this model were found to augment histamine synthesis in PMN isolated from the bone marrow. Rapid post-translational processing of HDC was observed in PMN, and the dominant form of HDC was the mature 53-kDa form, which was found to co-localize with a granule enzyme, matrix metalloproteinase-9 (MMP-9). Treatment of PMN with the phorbol ester PMA, which stimulates the release of MMP-9, did not liberate the granular HDC. Immunofluorescence studies using an anti-HDC antibody strongly suggested that HDC is bound to the cytosolic side of the granule membranes. These observations suggest that HDC is induced upon infiltration of PMN into the mouse peritoneal cavity and that histamine is synthesized by HDC attached to the granule membranes of PMN. PMID- 15114682 TI - CpG DNA redirects class-switching towards "Th1-like" Ig isotype production via TLR9 and MyD88. AB - Unmethylated CpG-containing DNA plays a critical role in immunity via the augmentation of Th1 but suppression of Th2 T cell responses. We describe here that CpG motifs also redirect isotype production by murine B cells to "Th1-like" Ig isotypes (IgG2a, IgG2b, and IgG3) while suppressing Th2 isotypes (IgG1 and IgE). Using genetically mutant B cells, we find that the IgG2a, IgG2b and IgG3 isotypes are transcriptionally regulated via the promotion of class-switching, in a manner critically dependent upon TLR9 and MyD88. Thus, CpG DNA redirects Ig isotype production by regulating the specificity of class-switch recombination. PMID- 15114683 TI - Peripheral CD4loCD40+ auto-aggressive T cell expansion during insulin-dependent diabetes mellitus. AB - The generation of auto-aggressive T cells involves failure of central or peripheral tolerance. We previously demonstrated that peripheral CD4(lo)CD40(+) T cells give rise to pathogenic T cells in the non-obese diabetic (NOD) model. Here we show that peripheral CD4(+)CD40(+) T cells from diabetic or pre-diabetic NOD mice induce insulin-dependent diabetes mellitus. Consistent with breach of peripheral tolerance, CD4(lo)CD40(+) T cells expand with age in NOD mice but not in MHC-matched non-obese resistant (NOR) or BALB/c controls. Suggestive of a causal role for CD40 in autoimmunity, blocking CD40-CD154 interactions early during NOD development prevents autoaggressive T cell expansion while promoting increases in CD4(+)CD25(+) regulatory T cells. Importantly, CD40 signals promote expansion of V alpha 3.2(+) and V alpha 8.3(+) T cells. Furthermore, peripheral V alpha 3.2(+)CD40(+) T cells induce diabetes in NOD.scid recipients while V alpha 8.3(+) T cells or V alpha 3.2(+)-depleted T cell populations do not. This is the first demonstration that primary T cells transfer disease with the kinetics of auto-aggressive T cell clones and that specific TCR V alpha expansion promotes diabetes. PMID- 15114685 TI - Hypertension guideline 2003 update. AB - OUTCOMES: Extensive data from many randomised controlled trials have shown the benefit of treating hypertension. The target blood pressure (BP) for antihypertensive management should be systolic BP < 140 mmHg, diastolic < 90 mmHg, with minimal or no drug side-effects. However, a lesser reduction will elicit benefit although this is not optimal. The reduction of BP in the elderly and in those with severe hypertension should be achieved gradually over 6 months. Stricter BP control is required for patients with end organ damage, co-existing risk factors and co-morbidity, e.g. diabetes mellitus. Co-existent risk factors should also be controlled. BENEFITS: Reduction in risk of stroke, cardiac failure, renal insufficiency and probably coronary artery disease. The major precautions and contraindications to each antihypertensive drug recommended are listed. RECOMMENDATIONS: Correct BP measurement procedure is described. Evaluation of cardiovascular risk factors and recommendations for antihypertensive therapy are stipulated. The total cardiovascular disease risk profile should be determined for all patients and this should inform management strategies. Lifestyle modification and patient education plays an essential role in the management strategy. Drug therapy: First line--low dose thiazide-like diuretics; second line--add one of the following: reserpine, or beta-blockers or ACE inhibitors or calcium channel blockers; third line--add another second line drug or hydralazine or alpha-blocker. The guideline includes management of specific situations, i.e. hypertensive emergency and urgency, severe hypertension with target organ damage and refractory hypertension (BP > 160/95 mmHg on triple therapy), hypertension in diabetes mellitus, etc. VALIDITY: Developed by the Working Groups established by the Executive Committee of the Southern African Hypertension Society with broader consensus meeting endorsement. The 2001 version was endorsed by the South African Medical Association Guideline Committee. The 2003 revisions were endorsed by the Executive Committee and a wider Working Group. PMID- 15114686 TI - [Introduction of information technology in the field of cancer therapy]. AB - The introduction of information technology in the field of cancer therapy will realize not only better mutual communication among staff and patients, but also standardize the care process. Secondary utilization of medical information, such as cost-income analyses, will also be possible, though there will still be some limitations due to immaturity of the present systems. PMID- 15114687 TI - [Chemical evaluation by cancer cell line panel and its role in molecular target based anticancer drug screening]. AB - Mechanism-based or target-based evaluation of chemicals is important in the discovery and development of anticancer drugs. A new scale for the mechanism oriented evaluation is acquired by creating a database of drugs that includes their activities concerning growth inhibition against a set of cancer cell lines and by employing a specific data-mining method (Paull KD, et al: J Natl Cancer Inst 81: 1088-1092, 1989). According to this principle, we have established a new system for drug evaluation using a panel of 39 human cancer cell lines, JFCR-39 Cell Line Panel. The JFCR-39 Cell Line Panel is a system combining a wet system (drug-sensitivity test) and a dry system (data base and its mining), and can predict the mechanism of action of chemicals. Therefore, it is useful in anticancer drug discovery. The JFCR-39 Cell Line Panel now plays an important role as a core drug evaluation system in the molecular target-based drug screening conducted by Screening Committee of New Anticancer Agents supported by Grant-in-Aid for Scientific Research on Priority Area "Cancer" from The Ministry of Education, Culture, Sports, Science and Technology, Japan. I outlined the JFCR 39 Cell Line Panel in this review. PMID- 15114688 TI - [Screening of protein kinase inhibitors]. AB - To screen the compounds that inhibit protein kinases, we used an assay system in which activities of protein kinase A, protein kinase C, src family protein tyrosine kinases and eukaryotic elongation factor 2 kinase were simultaneously detected on a single gel for the cytoplasmic protein kinases. For the receptor type protein kinases, EGFR and Flt-1 tyrosine kinases were examined. Here, we briefly described some of the protein kinase inhibitors that have been approved or are under clinical development, and present some novel inhibitors that were found in our screening system. PMID- 15114689 TI - [Screening of DNA topoisomerase inhibitors]. AB - It has been established that etoposide (ETP), adriamycin (doxorubicin: DOX) and irinotecan (CPT-11), efficacious antitumor drugs widely used in clinics, target DNA topoisomerases (topo) in vivo. The present report attempts to explain why topos are the good targets of anticancer drugs from the point of view of enzymology of the enzymes and cell cycle behavior of tumor cells. Thus, many candidate anticancer drugs targeting topos are being screened, and preclinical and clinical studies thereof are being conducted world-wide and in Japan. PMID- 15114690 TI - [Application of NGF/PC12 cell system for the screening of tubulin inhibitors]. AB - Microtubules are stiff polymers that extend throughout the cytoplasm of eukaryotic cells and regulate the location of intracellular compartments. Microtubules are formed by the polymerization of tubulin molecules, each of which is a heterodimer consisting of 2 closely related globular polypeptides: alpha tubulin and beta-tubulin. Many of the microtubule arrays are labile and turn over rapidly; they depolymerize and repolymerize continually in living cells. One of the most striking examples is the mitotic spindle, which forms after the cytoplasmic microtubules disassemble at the onset of mitosis. The mitotic spindle is the target of a variety of agents that act by interfering with the exchange of tubulin subunits between the microtubules and the free tubulin pool. Some of these agents can be used as anticancer drugs. We have recently developed a simple and sensitive screening method for tubulin inhibitors, to which we have successfully applied the nerve growth factor/PC 12 cell system. In this report, we describe the details of our screening method for tubulin inhibitors, by which we have analyzed several well-known and new compounds as model experiments. PMID- 15114691 TI - [Assay for inhibitory activity of histone deacetylase]. AB - Recently, it has become clearer that abnormal epigenetics is profoundly involved in many human diseases. Histone acetylation is one of the important mechanisms by which epigenetics is controlled. Reversible histone acetylation, which occurs at conserved lysine residues clustered near the amino-terminus of core histones, mediates changes in nucleosome conformation and transcriptional activity. Acetylation and deacetylation are catalyzed by specific enzyme families, histone acetyltransferases (HATs) and deacetylases (HDACs), respectively. Since aberrant histone acetylation has been linked to malignant diseases in some cases, HDAC inhibitors bear great potential as new drugs due to their ability to modulate transcription and to induce differentiation and apoptosis. Some of the HDAC inhibitors are currently under clinical investigations. Therefore, HDAC has been considered as a molecular target for cancer therapy. We are providing a service for evaluating the HDAC inhibitory activity of compounds that are sent to the screening committee. PMID- 15114692 TI - [Screening for inhibitors of tumor invasion into basement membranes]. AB - During the metastatic cascade, a tumor cell passes through several connective tissue barriers that consist of various adhesive molecules, such as fibronectin, laminin, collagens, and other glucoproteins and proteoglycans. Tumor invasion is a complex process involving cell adhesion, motility (migration), and the degradation of tissue barriers caused by the different proteases secreted by tumor cells. Therefore, understanding the invasion mechanism and the control mechanisms of the invasive properties of tumor cells may help in the development of anti-metastatic and anti-invasive therapies. We focused our attention on the functional molecules involved in the invasive process as targets to control tumor metastasis, and screened the inhibitors of tumor invasion into basement membranes. PMID- 15114693 TI - [Screening of antiangiogenic compound]. AB - Cancer cells promote angiogenesis early in tumorigenesis because cancer cells require oxygen and nutrients for their survival they need to be supported by blood vessels. The angiogenic switch is driven by several proangiogenic factors, vascular endothelial growth factor (VEGF), interleukin-8 (IL-8), basic fibroblast growth factor (bFGF) and others. These proangiogenic factors are released from not only cancer cells but also various stromal cells by various stimulations, mutations or inflammation. From our experimental results using human lung cancer cell lines, survival factors in each cancer cell line are different. This makes cancer therapy difficult. Various angiogenesis inhibitors have been developed to block tumor angiogenesis. However we need to think how and when to use angiogenesis inhibitors together with conventional chemotherapeutic agents. PMID- 15114694 TI - [Screening for apoptosis inducers]. AB - We carried out a screening for drugs that can induce apoptosis in human monocytic leukemia U937 cells. In the screening, we found that 8-nitrocaffeine induces cell death distinct from typical apoptosis. Morphological and biochemical analysis revealed that reactive oxygen species mediates the 8-nitrocaffeine-induced necrotic cell death. PMID- 15114695 TI - [Structure-activity relationship analysis]. AB - All samples for anticancer drug screening were classified according to their structural features and their structure-activity relationships were analyzed. Synthetic gymnastatin analogs including JCI: 11788 and JCI: 11786 altered their selectivity for protein kinase inhibition with the length of a fatty acid chain. Although a new inhibitor of tubulin depolymerization, JCI: 11578, displayed a high correlation to known tubulin binders, novel inhibitors of tubulin polymerization, JCI: 11534, JCI: 11675 and JCI: 11676, exhibited poor correlations to tubulin binders. JCI: 11403 and JCI: 11407 inhibited topoisomerase I selectively and appear to belong to a new family of topoisomerase inhibitors. They are expected to be important key compounds for structure activity relationship analysis as well as new lead compounds for anticancer drugs. PMID- 15114696 TI - [Estimation of level of enzyme induction of thymidine phosphorylase, dihydropyrimidine dehydrogenase, and thymidylate synthase in primary and recurrent breast cancer]. AB - Several enzymes are closely related with the mechanism of action of fluoropyrimidine (FP). Dihydropyrimidine dehydrogenase (DPD) which catalyzes 5 fluorouracil (5-FU), thymidine phosphorylase (TP), responsible for catalyzing doxifluridine to 5-FU, and thymidylate synthase (TS) were estimated for breast cancer. TP level determined by enzyme-linked immunoabsorbant assay (ELISA), DPD level by ELISA and catalytic assay and TS level by fluorodeoxyuridine monophosphate (FdUMP) binding assay were estimated for 210 specimens from 125 consenting patients with primary or metastatic breast cancer who gave consent. TS level of T1 was higher than that of T2-4 (p < 0.05). A high positive correlation was observed between TP and DPD in the same specimen. The ratio of simultaneously resected metastasis/primary and postoperative recurrent/primary never exceeded two. Enzyme level, generally decreased with or without interventing FP therapy. Enzyme level increase was frequently observed in a series of no-drug, non-FP, and FP therapy regimens. For selection of chemotherapy or estimation of chemosensitivity, TS can be used, with either TP or DPD at first surgery, and also with intervening chemotherapy after recurrence. PMID- 15114697 TI - [A late phase II clinical study of S-1 in patients with progressed, refractory breast cancer]. AB - A late phase II clinical study of S-1 against advanced or refractory breast cancer was done by 37 institutes in Japan. S-1 was administered twice daily at 80, 100 or 120 mg/body/day consecutively for 28 days followed by 14 days of rest (1 course). Eighty-three patients were enrolled and 81 were eligible for the study. The response ratio was 42.0% with 6 CR and 28 PR and its 95% confidence interval for the response was 31.1 to 53.5%. The median survival period was 910 days (95% confidence interval was 493-1, 083 days). The observed major adverse reactions (> or = grade 2) were as follows: hematological toxicities: leukopenia 21.0% (17/81), neutropenia 28.4% (23/81), erythropenia 4.9% (4/81); gastrointestinal toxicities: anorexia 9.9% (8/81), nausea and vomiting 12.3% (10/81), diarrhea 8.6% (7/81), stomatitis 1.2% (1/81), and fatigue 8.6% (7/81). The severe adverse reactions (> or = grade 3) were as follows; hematological toxicities: neutropenia 8.6% (7/81), anorexia 4.9% (4/81), fatigue 3.7% (3/81), nausea and vomiting 1.2% (1/81), diarrhea 1.2% (1/81), stomatitis 1.2% (1/81). Grade 4 adverse reactions (neutropenia and fatigue) were observed only in 1 patient. The ratio without hospitalization was 87.7%. These results strongly suggest the superior efficacy and safety of S-1 against patients suffering from advanced, refractory breast cancer. Therefore, S-1 may be a new therapeutic agent to prolong the survival period of breast cancer patients due to its high antitumor activity and low toxicity. PMID- 15114698 TI - [A pilot study of combined chemotherapy with paclitaxel, doxorubicin and cisplatin for endometrial cancer]. AB - A pilot trial of combined chemotherapy with paclitaxel, doxorubicin and cisplatin was conducted in patients with advanced endometrial cancer. Between June 2000 and March 2002 8 patients were treated with combined chemotherapy, consisting of paclitaxel, 135 mg/m2; doxorubicin, 30 mg/m2; and cisplatin, 50 mg/m2 (TAP therapy). Patients received 3 to 5 courses of TAP therapy every 4 weeks. The major adverse effect was myelosuppression. All patients had grade 3 or 4 neutropenia, but did not have any severe infection with uncontrollable fever. Only 1 patient discontinued additional therapy due to grade 3 thrombocytopenia after 3 cycles. Grade 2 neurotoxicity occurred in 5 patients, but grade 3 was not observed. Among 5 patients with measurable tumors, 4 achieved partial response and 1 had no change of tumor size, indicating a response rate of 80.0%. We found that TAP therapy was feasible with G-CSF support and shows potential for high efficacy in advanced endometrial cancer. PMID- 15114699 TI - [Paclitaxel plus carboplatin in ovarian cancer-comparison of adverse effects between monthly and weekly administration]. AB - Adverse effects of first-line combination chemotherapy performed with paclitaxel (PTX) and carboplatin (CBDCA) (TJ regimen) on 15 ovarian cancer patients who had had no prior chemotherapy with cisplatin (CDDP) were reviewed retrospectively according to National Cancer Institute common toxicity criteria. The M group (M) consisted of 7 patients treated with a total of 45 courses of the M-TJ regimen. Every 3-4 weeks, PTX was administered as a 3-hour infusion at the average dose level of 175 mg/m2/course on day 1 and CBDCA (targeted AUC = 6) was also administered on day 1. The W group (W) consisted of 10 patients who received a total of 49 courses of the W-TJ regimen. They were treated with PTX (80 mg/m2, 1 h, average dose level = 203 mg/m2/course) on day 1, 8 and 15, and with CBDCA (targeted AUC = 5) on day 1 every 4 weeks. Adverse events with grade 3 or above hematologic toxicity were oligochromemia (M: 24.4%, W: 22.4%), leukopenia (M: 55.6%, W: 40.8%), neutropenia (M: 84.4%, W: 61.2%) and thrombocytopenia (M: 17.8%, W: 8.2%). Grade 3 or above nonhematologic toxicity was not found in the W group, and anorexia (2.2%), nausea (2.2%), diarrhea (2.2%) and arrhythmia (2.2%) were developed only in the M group patients. Toxicity grades for neutropenia, arthralgia, myalgia and neuropathy were significantly lower in the W group. Based on the collected data, the W-TJ regimen is considered to be more effective than the M-TJ regimen for reducing the grade and occurrence of adverse events in ovarian cancer patients. PMID- 15114700 TI - [Paclitaxel, ifosfamide, nedaplatin (TIN) for patients with metastatic urothelial cancer]. AB - TIN (ifosfamide 1.5 g/m2 daily for 3 days, paclitaxel 175 mg/m2, and nedaplatin 70 mg/m2 on day 1) was administered to patients with metastatic urothelial cancer previously treated by platinum-based chemotherapy and repeated every 4 weeks. Four patients received maintenance therapy, which consisted of 5'-DFUR 800 mg/day orally for 12 weeks and 1 subsequent course of TIN. This therapy regimen was repeated for 2 years from initiation of TIN. Eleven of 12 patients (91.6%) demonstrated a major response (3 complete responses, 8 partial responses), with durations of response ranging from 3 to 20 months. Progression-free survival time was from 0 to 20 months (median 8 months). One-year progression-free survival rate was 45.8%. Overall survival time was from 2 to 20 months (median 10.5 months). One-year overall survival rate was 53.5%. Grade 3/4 hematologic toxicity involved neutropenia in 100% and thrombocytopenia in 33.3%. Febrile neutropenia was observed in 5 patients (41.6%). Grade 3 nonhematologic toxicity involved malaise in 15.3%. No patient discontinued this therapy because of complications. TIN is a potent, well-tolerated regimen for previously treated patients with urothelial cancer. PMID- 15114701 TI - [Subset analysis of data in the Japanese patients with NSCLC from IDEAL 1 study on gefitinib]. AB - The multinational, multi-institutional clinical Phase II trial of gefitinib monotherapy, IDEAL (IRESSA Dose Evaluation in Advanced Lung Cancer) 1, included Japanese and non-Japanese patients with advanced non-small-cell lung cancer (NSCLC) pretreated with one or more chemotherapy regimens, at least one including platinum. To investigate whether survival is affected by gender or histological type of cancer, a retrospective, exploratory subset analysis was conducted including only Japanese patients from IDEAL 1 (n = 102 in total, 51 each in 250 and 500 mg/day groups). The median survival time of the 102 patients was 12.0 months and the one year survival rate was 50%. The median survival time was 13.8 months for the 250 mg/day group and 11.2 months for the 500 mg/day group and the one-year survival rate was 57% and 45% respectively. Survival was longer in patients with adenocarcinoma than those with other histological types of cancer, and was longer in those with symptom improvement than without. The median survival time in females was longer than that in males. The results suggest that gefitinib could be superior to classical anticancer agents with regard to not only the response rate but also survival time in patients with NSCLC, particularly adenocarcinoma, previously treated with chemotherapy. Further studies are needed to identify factors affecting survival. PMID- 15114702 TI - [A case of Barrett's esophageal carcinoma successfully treated with TS-1 in an elderly patient]. AB - A 92-year-old man was admitted to the hospital for rehabilitation. Upper gastrointestinal endoscopy was performed 2 weeks after admission because of vomiting which was found to be due to a Barrett's esophageal carcinoma at the lower esophagus. We chose chemotherapy with TS-1 at the dose of 50 mg/day in consideration of his age and performance status. His difficulty in swallowing disappeared rapidly. After 7 days of treatment, grade 3 anorexia appeared, so the dosage of TS-1 was reduced to 25 mg/day. After the reduction of TS-1, anorexia soon disappeared. The regimen consisted of TS-1 every 6 weeks as 1 cycle. After 7 cycles of treatment, upper gastrointestinal endoscopy and upper gastrointestinal series revealed that the tumor had a completely disappeared. The effects of chemotherapy were judged as CR. This case suggests that administration of TS-1 is an effective choice of chemotherapy not only for gastric carcinoma but also Barrett's esophageal carcinoma. PMID- 15114703 TI - [A case report of advanced cardiac cancer showing a complete response to TS-1 as neoadjuvant chemotherapy]. AB - A 69-year-old female had complaints of vomiting, appetite loss and feeling of pharyngeal obstruction. She was diagnosed with a 3'-shaped advanced cardiac cancer with esophageal invasion. A biopsy revealed poorly differentiated adenocarcinoma. The tumor was T3 (SE) N2, Stage IIIB indicating a poor prognosis. After informed consent, TS-1 was administrated as preoperative chemotherapy. Chemotherapy with TS-1 was very effective, and the tumor noticeably decreased. Next, total gastrectomy was performed. Histopathological findings revealed that the primary tumor and lymph node had become scarred and fibrous, indicating a complete response (Grade 3). In the future, TS-1 can be expected to display efficacy in neoadjuvant chemotherapy for patients with advanced gastric cancer who have poor prognoses. PMID- 15114704 TI - [A case of advanced gastric cancer effectively treated by combined chemotherapy of paclitaxel (TXL) and CDDP]. AB - The patient was a 73-year-old man with unresectable advanced gastric cancer and celiac and supraclavicular lymph node metastases. Neoadjuvant chemotherapy consisting of paclitaxel (TXL) and CDDP was administered. TXL (80 mg/m2) and CDDP (25 mg/m2) was administered weekly on day 1, 8 and 15 as 1 cycle. After 4 cycles of TXL/CDDP administration, the lymph node metastases and gastric tumor had decreased almost completely in size and distal partial gastrectomy was performed. After surgery, the patient was treated with 4 courses of TXL/CDDP and has survived without recurrence to the present. TXL/CDDP is associated with few adverse events in hospital visits, and is thought to be an effective chemotherapy against advanced gastric cancer. PMID- 15114705 TI - [A case of gastric cancer with multiple liver metastases responding completely to TS-1]. AB - We report a case of a 65-year-old male with stage IV gastric cancer accompanied by liver metastases, which showed a significant response after administration of TS-1. One hundred and twenty mg/body/day of TS-1 was orally administered without hospitalization. After 3 months, upper GI endoscopy showed improvement of primary gastric lesion, and cancer cells could not be detected under biopsy. After 2 months, computed tomography (CT) showed a reduction in the multiple liver metastases. Moreover, after 15 months, CT showed a complete regression of the multiple liver metastases, for a complete response (CR). The serum level of carcinoembryonic antigen (CEA) was reduced from 115 to within normal range. Noticeable critical adverse effects did not appear. Treatment on an outpatient basis, therefore, greatly contributed to his quality of life. We judged that TS-1 might be a candidate anti-cancer drug for first-line chemotherapy for advanced gastric cancer. PMID- 15114706 TI - [A case of complete response in a primary lesion treated by combined chemotherapy of TS-1 and CDDP for small cell carcinoma of the stomach with liver metastasis]. AB - A 75-year-old male patient with small cell carcinoma of the stomach and liver metastasis was treated by combined chemotherapy of TS-1 and CDDP. One course consisted of TS-1 (120 mg/day) administered for 14 days followed by 14 days rest. CDDP (108 mg/day) was administered by 24-hour continuous intravenous infusion at day 8 after the start of TS-1. After 3 courses, endoscopic examination revealed complete disappearance of the primary tumor with no cancer cells detected by endoscopic biopsy. CT-scan showed that the metastasis of the left lobe of the liver had disappeared and also that the metastasis of the right lobe of the liver was remarkably reduced (75%). The primary lesion was estimated CR, the metastasis PR, and the synthesis PR. The TS-1/CDDP chemotherapy regimen is considered effective for small cell carcinoma of the stomach with liver metastasis. PMID- 15114707 TI - [A case of advanced gastric cancer successfully treated with neoadjuvant chemotherapy of combined weekly paclitaxel and doxifluridine (5'-DFUR)]. AB - We report a case of advanced gastric cancer with metastasis to the paraaortic lymph nodes that showed a remarkable response to treatment with a combination of weekly paclitaxel and doxifluridine (5'-DFUR). The patient was a 72-year-old man. Oral chemotherapy of TS-1 was discontinued due to drug induced eruption. Alternatively, we administered weekly paclitaxel/5'-DFUR combination therapy. Paclitaxel was infused at a dose of 100-130 mg after short premedication and continued for 2-3 weeks with a 1 week rest. 5'-DFUR was administered orally at a dose of 800 mg/day for 14-21 consecutive days. After 4 courses of this therapy, the primary carcinoma and lymph nodes decreased in size (PR). Consequently, the patient underwent a total gastrectomy with paraaortic lymph node dissection, which resulted in a curative resection of the cancer cells macroscopically. Except for afebrile neutropenia (grade 4), no major adverse reactions were observed. Histological examination revealed that the cancer cells were degenerated to a moderate extent. Weekly paclitaxel/5'-DFUR combination may be a promising regimen for patients with advanced gastric cancer as preoperative chemotherapy. PMID- 15114708 TI - [Examination of the feasibility of TS-1 for postoperative advance stomach cancer patients]. AB - AIM: The purpose of this study was to assess the safety of TS-1 when used as adjuvant chemotherapy after surgical resection of advanced stomach cancer. SUBJECTS: The subjects were 20 patients with stage III or IV, curability B resected gastric cancer. METHODS: The methods consisted of analysis of background factors, assessment of the administration and dosage of TS-1, and associations with adverse reactions. RESULTS: Mean age was 62.8 years, and the histological type was differentiated in 7 cases and undifferentiated in 13 cases. There were 4 stage IIIA cases, 10 stage IIIB cases, and 6 stage IV cases, and the extent of gastric resection consisted of gastrectomy in 8 cases and total gastrectomy in 12 cases. TS-1 administration was started an average of 57 days postoperatively. The dosage regimen was 4 weeks on and 2 weeks off in 10 cases, and different regimens were used in the other 10 cases. The dose of TS-1 was the recommended dose in 9 cases and a reduced dose in 11 cases, and the mean number of courses was 2.8. Adverse reactions occurred in a total of 14 cases. Digestive system toxicity developed in 5 cases and hematological toxicity in 12. No grade 3 or 4 toxicity was observed. Treatment could be continued in 18 cases. The occurrence of adverse reactions tended to be concentrated immediately after administration of TS-1. CONCLUSION: TS-1 was safely used as adjuvant chemotherapy after resection of advanced stomach cancer. PMID- 15114709 TI - [Complete response of gastric cancer with interaorticocaval lymph node recurrence to weekly administration of paclitaxel--a case report]. AB - We report a case in which weekly administration of paclitaxel produced a complete response for gastric cancer with interaorticocaval lymph node recurrence. A 55 year-old man who underwent total gastrectomy for advanced gastric cancer had pathological findings of tub2, se, n2, ly2, v2, stage IIIB. As an outpatient, he was first treated with TS-1 (100 mg/day) for 5 cycles. CEA increased gradually however, and 7 months postoperatively he was found to have interaorticocaval lymph node swelling in upper abdominal CT and was diagnosed with a recurrence of gastric cancer. He underwent weekly administration of paclitaxel as second-line chemotherapy. Paclitaxel was administered by 1-hour intravenous infusion at a dose of 90 mg/body weekly after short premedication as an outpatient. This was continued for 3 weeks followed by 1 week of rest. CEA decreased gradually, and the swollen lymph node had responded completely after 10 cycles. The only toxic side effect was alopecia (grade 1). No major adverse effects such as hypersensitivity, leukopenia or peripheral neuropathy were observed. PMID- 15114710 TI - [Weekly administration of paclitaxel as a second-line chemotherapy for advanced and recurrent gastric cancer]. AB - Chemotherapy with TS-1 has recently become the first-line chemotherapy for recurrent and unresectable gastric cancer in Japan. Therefore, the establishment of a second-line chemotherapy is needed for cases that show resistance and aberrant effect to TS-1. In this study, 7 patients were treated with weekly administrations of paclitaxel after TS-1 treatment. We assessed the therapeutic effect and feasibility of chemotherapy with weekly administration of paclitaxel. Our results showed that weekly administration of paclitaxel could be a promising regimen as a second-line chemotherapy after TS-1. PMID- 15114711 TI - [Report of two cases with pleural effusion and ascites that responded dramatically to the combination of thalidomide, celecoxib, irinotecan, and CDDP infused in thoracic and abdominal cavities]. AB - Malignant pleural effusion (PE) and ascites are associated with highly symptomatic, advanced-stage cancers. These fluid accumulations cause severe symptoms such as abdominal distention, shortness of breath, cachexia, anorexia, and fatigue. Malignant PE and ascites have consistently been shown to indicate a poor prognosis in advanced-stage cancer patients, being associated with high morbidity and mortality. The efficacy of this treatment is variable and does not prolong the survival of cancer patients. Clearly, a more effective therapy for malignant PE and ascites is needed. Vascular permeability factor (VPF) from malignant ascites and PE have been hypothesized to be responsible for the fluid accumulations. In addition, malignant PE and ascites contain high levels of biologically active VEGF. VEGF was discovered as a potent angiogenesis stimulator and recognized to be VPF. Increased amounts of COX-2 have been detected in epithelial and stromal cells and COX-2 in mammary tissue is sufficient to induce cancer. It is suggested that COX-2 stimulates angiogenesis. A combination of molecular target inhibitors (thalidomide and celecoxib) and standard cytotoxic drugs appear to increase efficacy of each drug, decrease the side effects of cytotoxic drugs and prolong life. PMID- 15114712 TI - [A case of ovarian cancer stage IV and advanced rectal cancer responding to paclitaxel/carboplatin]. AB - A 43-year-old woman who had ovarian cancer with massive ascites and pleural effusion as well as rectal cancer underwent probe laparotomy. However, only bilateral adnexectomy was performed since radical surgery was impossible due to severe carcinomatosa peritonitis. Three courses of chemotherapy with paclitaxel and carboplatin (T-J therapy) were carried out. Following chemotherapy, the ascites and pleural effusion had completely disappeared and the size of rectal cancer had shrunk as well. The chemotherapeutic effect was evaluated as a partial response in the rectal cancer. Therefore, as interval debulking surgery, abdominal total hysterectomy, omentectomy, low anterior resection pelvic lymphadenectomy were performed. Three further courses of chemotherapy were performed. There is no evidence of recurrence of ovarian cancer and rectal cancer 3 years after the first surgery. PMID- 15114713 TI - [Case report of two colorectal cancer patients with liver metastasis showing favorable response to fluorouracil and l-leucovorin therapy]. AB - We treated 2 patients with colorectal cancer accompanied by liver metastasis who showed favorable response to combined treatment with fluorouracil and l Leucovorin. Case 1 was a 40-year-old man with rectal carcinoma (moderately differentiated adenocarcinoma; se, n1, p1, H0, stage IV). He underwent low anterior resection of the rectum and postoperatively showed an increase peritoneal dissemination and liver metastasis. The patient was given 600 mg/m2 fluorouracil and 250 mg/m2 l-Leucovorin concomitantly for 7 courses. At course 3, the liver metastasis showed partial response and, during the 18 months after the 7th course, his peritoneal dissemination and other lesions were well controlled. As of 30 months post-treatment the patient was alive. Case 2 was a 60-year-old man with rectal carcinoma (well differentiated adenocarcinoma; ss, n1, p0, H1, stage IV). He underwent low anterior resection and postoperatively was given 3 courses of fluorouracil and l-Leucovorin. At course 2, he showed complete response, and, at present 8 months after the 2 courses, the patient continues to show complete response. Adverse drug reactions in both patients were controlled on an outpatient basis. While in the past liver metastasis has been treated by hepatic arterial infusion chemotherapy, a combination therapy with fluorouracil and l-Leucovorin can be used on an outpatient basis and results in a favorable response. This suggests that this combination therapy has clinical significance. PMID- 15114714 TI - [Weekly paclitaxel therapy for locoregional lymph node recurrence of estrogen receptor-negative breast cancer]. AB - We treated 7 patients with locoregional lymph node recurrence of estrogen receptor-negative breast cancer with weekly paclitaxel (TXL) therapy. TXL was administered by 30-minute infusion at a dose of 80 mg/m2 after short premedication every week on an outpatient basis. Administration was continued for 3 weeks with a 1 week rest. Five patients had received prior anthracycline therapy. No patients had received prior therapy for recurrence. The median number of cycles was 4.9 cycles (range, 3-6 cycles). The overall response rate was 84% among 5 complete responders and 1 partial responder. Four of 5 complete responders relapsed after the completion of weekly TXL therapy. Grade 2 leukopenia occurred in 3 patients, and grade 1 peripheral neuropathy occurred in 1 patient. No patients had a serious side effect. Weekly TXL therapy is effective and well tolerated in patients with locoregional lymph node recurrence of estrogen-receptor-negative breast cancer, but the problem of recurrence to other sites after treatment remains to be solved. PMID- 15114715 TI - [A case of neck metastasis from hypopharyngeal carcinoma successfully treated with TS-1]. AB - We report a case of hypopharyngeal carcinoma with recurrent neck metastasis. Preoperative chemoradiation and pharyngo-laryngo-esophagectomy with bilateral neck dissection was done as first-line therapy. Six months later, the carcinoma metastasized to the right paratracheal lymph node. Because of resistance to chemoradiation (CBDCA) as second-line therapy, we administered TS-1 for the treatment after 60 Gy radiation. The chemotherapy with TS-1 resulted in complete response by diagnostic imaging (CT). There was no sign of recurrence for 1 year after 5 courses of treatment. TS-1, which allows oral chemotherapy on an outpatient basis, would be a useful drug for treatment with radiotherapy in patients with advanced and/or recurrent head and neck cancer. PMID- 15114716 TI - [An advanced case of squamous cell carcinoma in the left buccal mucosa, upper gingiva, and maxillary sinus (T4N0M0) showing a complete response to chemotherapy with TS-1]. AB - We report a case of advanced squamous cell carcinoma in the left buccal mucosa, upper gingiva, and maxillary sinus (T4N0M0) showing a complete response to oral chemotherapy with TS-1. The patient was an 89-year-old female with severe dementia. We carried out chemotherapy with TS-1 50 mg/day, without surgical treatment. The tumor disappeared clinically at 4 months after 3 courses of the TS 1 administration. Adverse drug reactions, including vomiting, leukopenia and thrombopenia, forced a stop of the administration of TS-1. Although she finally died of in senescence 2 months from the cease of administration, there was no recurrence of the cancer at the time. PMID- 15114717 TI - [Oral cancer]. AB - This paper reviews the Japanese literature regarding sentinel lymph node localization in head and neck surgery and relevant domestic and foreign articles from other fields of medicine, and reports our results. Even though we have less experience with it in Japan, we believe the sentinel node concept for the head and neck region will be established. Further investigation and practical application in clinical settings are anticipated in the future. PMID- 15114718 TI - [Anti angiogenesis]. AB - Based on presentations on the basic concepts and scientific rationale of anti angiogenic approaches to cancer therapy and the possible applications in the area of prostate cancer, gastrointestinal cancer, lung cancer and breast cancer it is easy to conclude that development of anti-angiogenic approaches into clinical therapies is extremely challenging. It is now well established that cancer growth is increased by angiogenic factors and that inhibition of angiogenesis decreases growth and metastatic potential. Anti-angiogenic effect can be obtained through interference with multiple targets. Further development of new strategies involving such novel cancer therapies requires wide reaching development of translational research abilities. However, for moving new therapies into the clinic same rigorous criteria need to be applied as is done for traditional therapies. Angiogenesis appear to be a critical factor for development of prostate, gastric, lung and breast cancers. Development of new anti-angiogenic treatment modalities might become very important in these diseases. A critical requirement for the successful clinical development will be the development of imaging techniques that can help evaluate the effect on blood vessel functionality. Such surrogate markers of anti-angiogenic effect will be essential for optimising molecules and doses. PMID- 15114719 TI - [Combined antineoplastic agents guidelines--guidelines for clinical trials I/II]. PMID- 15114720 TI - Dietary intake of selected minerals for the United States population: 1999-2000. AB - This report presents dietary intake estimates for selected minerals from the National Health and Nutrition Examination Survey, 1999-2000, for the U.S. population. These minerals included: calcium, copper, iron, magnesium, phosphorus, potassium, selenium, sodium, and zinc. Mineral intakes are estimated from one 24-hour dietary recall interview. Population means, medians, and standard errors of the mean are weighted to produce national estimates, and are presented by sex and age groups. Assessment of dietary intakes is an important part of monitoring the nutritional status of the U.S. population. PMID- 15114721 TI - [A new method of macrofragmentation of dense nuclei by using loop fragmentizer]. AB - The study results showed a principle possibility to fragmentize the most dense lens nuclei by means of a nylon cutting loop. The safety and efficiency of applying a loop fragmentizer in phacoemulsification of high-density cataract were equally demonstrated. The method advantages are conditioned by that a preliminary splitting of the nucleus into fragments without trenches being shaped cuts essentially the total capacity and duration of ultrasound impact, thus, the irrigation time and the needed volume of balance salt solution also decrease intraoperatively: it, naturally, diminishes the generally-known traumatic aftermaths of surgical intervention. PMID- 15114722 TI - [Cell loss in the posterior corneal epithelium after cataract extraction by Nd:YAG laser with wave length 1.44 mcm]. AB - The purpose of the case study, as described in the paper, was to evaluate the cell loss in the corneal posterior epithelium (CPE) after laser cataract extraction (LCE) by means of a new laser unit designed on the basis of ND:YAG laser with a wave length of 1.44 microns. The conditions of 59 eyes with high (degrees 3-4) and extra high (degree 5) lens density were analyzed preoperatively and 3 months after LCE. Group 1 comprised 51 surgical cases, which were ideal in the technical respect; Group 2 comprised 8 surgical cases involving an intraoperative short-time lifting of the fragmentized lens mass into the eye anterior chamber during aspiration. The method of contactless endothelial biomicroscopy ("SP-1000" unit, "Topcon Co.", Japan) was made use of. According to the obtained results, it can be stated: the CPE cell loss made 1-9% (4.69 +/- 0.7%) and 1-18% (9.4 +/- 8.4%) in Groups 1 and 2, respectively. The total energy exposition level to laser radiation is the most significant factor affecting the CPE cell loss. The pulse energy radiation level did not affect the CPE cell loss. The corneal posterior surface was subject to a highly adverse impact in the fragmentation of brown cataracts. Finally, the results are indicative of an insignificant surgical trauma in LCE. PMID- 15114723 TI - [Limbal transplantation in the treatment of and optic rehabilitation of the patients with severe chemical burns of the eyes]. AB - Described in the paper are the results of observations of the autogenous and allogenic limb transplantation (LT) made in 4 patients at exacerbation of severe and extra severe burns of the eyes and made in 8 patients in the remote period after severe chemical burns (4 months and more after trauma). A total vascularized corneal leukoma shaped up in 3 patients with LT made at exacerbation; an intense corneal opacification was observed in 1 more such patient. As for the patients, who were operated on in the remote period, better corneal properties and significantly improved visual acuity were registered in them. Relapsing corneal conjunctivalization were observed only in 3 cases of allogenic LT. Thus, LT was proven to be an effective tool in the remote visual rehabilitation of patients with severe eye burns, whereas, the procedure made at exacerbation and in cases of acute and extra acute burns with extensive lesions in the conjunctiva is low effective or a failure. PMID- 15114724 TI - [The parameters of the body immune reactivity in applying the therapeutic keratoplasty for the treatment of ocular burn lesions of extra severity]. AB - One hundred and two (128 eyes) patients with stage 4 ocular burns underwent examinations. One-stage therapeutic keratoplasty was used in 67 (67.7%) patients. Retransplantation was made in 35 (34.3%) of patients due to graft rejection. According to the study, the more severe the clinical course of burn lesions is (cases that necessitated transplantation in the present research), the more pronounced changes are observed in the body immune reactivity of victims. In particular, the below peculiarities were observed: a progressing reduction of the contents of lymphocytes' and of B-lymphocytes; and a diminishing phagocytic activity of neutrophils in the peripheral blood registered at the terminal treatment stage in the patients after retransplantation versus the 1st-group patients with a more favorable clinical course. The compensatory increase of T lymphocytes, T-helpers and of IgA, IgG was found to be less pronounced in group 2. A notably more intense sensitization response of the body to corneal antigens was established in the patients after retransplantation, which was a basis for using this index as a prognostication test in respect to the graft rejection crisis. PMID- 15114725 TI - [Neuro-ophthalmological and radiological signs of Aicardi syndrome]. AB - Described in the paper are the results of ophthalmologic, ultrasound and radiological examinations of 3 girls with the Aicardi syndrome. It was for the first time, that an expansion of the optic-nerve disk excavation was described in patients with the above syndrome; such expansion was conditioned by trans synaptic neuron degeneration induced by lesions to optic radiation in view of prenatal of encephaloplastic processes. The changing brain white substance as projected by optic radiation was detected in all children by computer and magnetic resonance tomography. Besides, other anomalies of the optic nerve (bindweed syndrome, coloboma) and of the retina are elucidated. Finally, the pathogenesis and differential diagnostics of the Aicardi syndrome are under discussion. PMID- 15114727 TI - [Accommodation in the implantation of phakic intraocular lenses for the correction of high myopia]. AB - Implantations of 67 phacic intraocular lenses (IOL) in myopia ranging from 8 to 17 d were analyzed. The postoperative refraction was -0.97 +/- 0.45 d. The accommodation response, as a sharp reaction, and accommodation tracing were investigated in all patients. The "Nidek" AA2000 lens accommodograph was made use of. An inhibition and a higher threshold in the rapid accommodation response were registered in 86% of cases, tonic phase inhibition was found in 72% of cases and a limited maximal amplitude was noted in 45% of cases in accommodation tracing performed within examinations made 1 week after surgery. Inhibited accommodation response (86%), and stage-type relaxation in accommodation tracing (49%) were stated in examinations 3 weeks later. The implantation of phacic IOL for the correction of high myopia can bring about an accommodation dysfunction. The accommodation time of the "accommodation-IOL" system ranges from 1 to 3 months. Such adaptation is associated with an intensified accommodation reflex. PMID- 15114726 TI - [Changing electrophysiological parameters in the eye in children with insulin dependent diabetes mellitus]. AB - Fifty-eight (58) children with insulin-dependent diabetes mellitus (IDDM) were examined and shared between 6 groups: Group 1--primarily diagnosed diabetes mellitus (DM); Group 2--DM with an up to 5-year history; Group 3--DM with a an up to 10-year history; Group 4--children with non-proliferative diabetic retinopathy; Group 5--children with diabetic cataract. Forty-five healthy children were in the control group. According to the pattern registration of visual induced potentials (RVIP), a reliable prolongation of latency P100 was detected in children of Groups 1 and 4. The results of the general electroretinography (ERG) showed a reliably decreased amplitude of the basic wave b- in Groups 1-4 and an increased amplitude of wave a- in Groups 4 and 5. According to photopic ERG, a reliably decreased amplitude of wave a- was detected in Groups 1-5 and a decreased amplitude of wave b- was registered in all groups except for Group 2. Scotopic ERG showed, in Group 5, an increased amplitude of wave a- (p < 0.05) at the normal values of wave b-, and, in Groups 1-4,--a significantly inhibited amplitude of wave b-. Finally, rhythmic ERG showed a reliably inhibited amplitude in all groups. PMID- 15114728 TI - [A comparative analysis of hemodynamic parameters within the evaluation of the glaucomatous process in patients with myopia]. AB - One hundred and eighty (180) persons (363 eyes), mean age 63.7 +/- 5.2, mean follow-up--6.1 +/- 1.2, were investigated. Control group 1 comprised 46 patients (92 eyes) with emmetropia and different-degree myopia; control group 2 had 74 persons (148 eyes) with primary open-angle glaucoma (POAG). The main group had 60 patients (119 eyes) with POAG combined with myopia. The ocular hemodynamics parameters were found to reliably differ, in patients with myopia, without glaucoma and with the anterior-posterior eye axis of 25 mm, from the similar parameters in persons with emmetropia. The process aggravates in myopic patients with glaucoma due to reliably worsened ocular hemodynamics parameters and due to a higher intraocular pressure without any reliably changing systemic hemodynamics. The measuring of ophthalmoplethysmography parameters could be handy in examining the POAG patients with myopia in order to detect if glaucoma is progressing or not. PMID- 15114729 TI - [The anatomic and functional specificity of the superior tarsal Mueller's muscle]. AB - The anatomic-and-functional specific features of the structure, topography and of spacious architectonics of the superior tarsal muscle were detected and morphologically confirmed within a case study performed on 16 cadaverous fragments of the superior eyelids. The research of functional biomechanics of the superior eyelids made in 220 healthy subjects made it possible to expand the criteria used in evaluating the results of adrenergic stimulation of unstriped muscles, which must be considered as prognostic criteria in surgical treatment; an appropriate morphometric substantiation is suggested. PMID- 15114730 TI - [A study of total IgE in inflammatory pathologies of the eye anterior segment]. AB - The paper contains data on comparative research of the IgE content in blood serum and lachrymal fluid in patients with ophthalmoherpes and with other inflammatory diseases of the eye. A higher IgE level was found in blood serum and lachrymal fluid in cases of ophthalmoherpes as well as in lachrymal fluid in cases of allergic, Chlamydia and fungus diseases of the eye. The data obtained can be used in the diagnostics of allergic eye diseases as well as in elaborating complex treatment methods for herpetic, Chlamydia and fungus lesions of the anterior eye segment. A detection of the local allergenic effect of acaricide drugs exerted on the conjunctiva and eyelids makes it obligatory to consider the above fact while treating patients with demodectic blepharoconjunctivitis. PMID- 15114731 TI - [Epidemiological aspects of basal cell cancer of the eyelids' skin in the Cheliabinsk region]. PMID- 15114732 TI - [A technique of reconstructing of absent eyelids]. PMID- 15114733 TI - [The angle status of the anterior chamber in primary closed-angle glaucoma with respect to a disease stage and to choosing a treatment scheme]. PMID- 15114734 TI - [Ocular cysticercosis in India]. PMID- 15114735 TI - [A case of gonorrheal conjunctivitis complicated by corneal ulcer and uveitis]. PMID- 15114736 TI - [Silicone tamponade in modern surgery of retinal detachment]. PMID- 15114737 TI - [An analysis of potentialities of modern morphofunctional methods of retinal examination in cataract surgery]. PMID- 15114738 TI - [Current aspects of tuberculosis uveitis]. PMID- 15114739 TI - [Significance of transport adenosinetriphosphatase in eye pathology]. PMID- 15114740 TI - [The specificity of calculating the force of intraocular lens after keratorefraction surgery]. PMID- 15114741 TI - [Professor A. G. Liutkevich--founder of the Eye Clinic of Voronezh Medical University and of the "Annals of Ophthalmology" journal]. PMID- 15114742 TI - [The results of long-term permixon treatment in patients with symptoms of lower urinary tracts dysfunction due to benign prostatic hyperplasia]. AB - The trial enrolled 155 patients (mean age 65 years) with documented benign prostatic hyperplasia and lower urinary tracts symptoms (LUTS) (IPSS > 6). All the patients received permixon in a dose 160 mg twice a day for 2 years. The data on 130 patients eligible for assessment were processed statistically by dynamics of IPSS, quality of life (QOL), index of sexual function (MSF-4), size of the prostate, urodynamic and biological parameters which were estimated in 6 (V6), 12 (V12), 18 (V18) and 24 months (V24). Clinical examination with registration of all side effects was made each 3 months. Permixon was found to noticeably reduce IPSS and QOL and increase maximal urine flow speed. The size of the prostate diminished insignificantly. Sexual function remained unchanged for 1 year and improved markedly within the second year (p = 0.001). Permixon had no effect on the level of prostate-specific antigen. Plasma hormones (testosterone, DHT, estradiol, LH, androstendion) did not change. Nine patients developed 10 side effects but they were unrelated to the treatment. PMID- 15114743 TI - [Proximate and remote results of ureterosigmoidostomy with creation of Mainz pouch II and Hasan reservoirs]. AB - The aim of the study was analysis of short- and long-term results of treating patients operated with intestinal replacement of the urinary bladder by Mintz pouch II and Hasan. Thirty-two ureterosigmostomies with creation of the reservoir from the rectosigmoid angle (17 Mintz pouch II and 15 Abol-Enein operations) were made from 1997-2002. The age of the patients ranged from 16 to 73, the follow-up was 12-72 months. The operation was indicated in infiltrative cancer of the urinary bladder, microcystis of various etiology, extrophy of the urinary bladder. Postoperative urine retention was observed in all the patients both in the day and night time. Renal function remained unchanged or improved in 93% patients. Quality of life was good. All the patients except two with recurrent attacks of pyelonephritis were socially adapted. Able patients resumed their jobs. Early after the operation one woman developed incompetence of the sutures of the reservoir anterior wall which required relaparotomy. Four patients died of local recurrence and/or distant metastases in different terms after surgery. Two patients had reflux with frequent attacks of pyelonephritis as well as a serious impairment of acid-base balance manifesting as hyperchloremic acidosis, the other two patients had late unilateral strictures of the intestinoureteral anastomosis which was reestablished. Thus, updated ureterosigmostomy significantly reduces the rate of complications and is a good alternative to heterotopic and ortotopic urine derivation in contraindications for the operations. Moreover, Hasan's operation allows implantation of delated ureters. In most cases, ureterosigmostomy with formation of reservoir from rectosigmoid angle provides good quality of life in adequate selection of patients. PMID- 15114744 TI - [Resistance of ambulatory urinary infection pathogens according to the data of multicenter microbiological studies UTIAP-I and UTIAP-II]. AB - AIM: To study causative agents and sensitivity of E-coli strains isolated from adult outpatients with uncomplicated urinary infection (UI) in different regions of Russia. MATERIAL AND METHODS: A multicenter prospective epidemiological study included adult patients with uncomplicated infections of the upper or lower urinary tracts. MPK of antibiotics was established by dilution in agar according to NCCLS recommendations, 2000-2002. RESULTS: Among UI causative agents, E. coli was most frequent (85.9%). K. Pneumoniae, Proteus spp., Staphylococcus spp., P. Aeruginosa, Enterococcus spp. occurred much less frequently (6, 1.8, 1.6, 1.2, and 1.0%, respectively). E. Coli UI was highly resistant to ampicilline (37.1%), cotrimoxasol (21%), maximal resistance being in St-Petersburg (51.9 and 31.5%, respectively). Such oral antibiotics as norfloxacin and ciprofloxacin, cefuroxim, amoxicillin/clavulanat, nitrofurantoin were maximally active against E. Coli (4.3, 2.4, 2.6 and 1.2%, respectively). CONCLUSION: High resistance of E. coli, which is the chief causative agent of uncomplicated UI, to ampicillin, cotrimoxasol was detected. Fluoroquinolones, amoxicillin/clavulanat, nitrofurantoin, cefuroxim have high microbiological activity. On the basis of the pharmacokinetic, safety and other evidence it is concluded that drugs of choice for therapy of uncomplicated UI in Russia are oral fluoroquinolones. PMID- 15114745 TI - [Diagnostic errors in purulent pyelonephritis]. AB - Most frequent errors in diagnosis of purulent pyelonephritis have been analysed. It is shown that only a total of data obtained on the disease history, clinical symptoms, results of laboratory tests, ultrasound or x-ray symptoms of purulent pyelonephritis enable making an accurate diagnosis and choice of adequate treatment policy. PMID- 15114746 TI - [Combined therapy of interstitial cystitis using the "Aeltis-Synchro-02-Iarilo" device]. AB - Multiple modality therapy of interstitial cystitis (IC)--the disease characterized by nicturia, pelvic pains, imperative pollakiuria--is considered. As IC nature is not well known, its treatment remains empiric. Among the underlying causes, most probable are autoimmune, allergic, infectious, neurological, vascular. Therefore, the treatment should be multi-modality. Most usable now is combined chemotherapy. Perspective is also IC treatment with medicines in combination with physiotherapy (electromagnetolaser AELTIS-SYNCHRO 02-YARILO"). Endovesical electrophoresis can be also applied. PMID- 15114747 TI - [The combined treatment of interstitial cystitis in women with spasmex]. AB - Efficiency of interstitial cystitis (IC) treatment of female outpatients is shown. Combined treatment of IC was supplemented with the drug spasmex which has a peripheral anticholinergic and direct antispastic actions. It quickly relieves irritative symptoms, improves quality of life, shortens temporary disability and lowers the cost of the treatment. PMID- 15114748 TI - [Organ-saving treatment of penile cancer]. AB - Conservative penis-salvage treatment provides a complete local effect in 55% cases, preservation of the penis in 51.4% patients without a fall in long-term specific and recurrence-free survival compared to penectomy and can be recommended as an alternative to penis amputation in patients with stages Tis-T2. Removal of a penile tumor raises efficacy of salvage treatment and insignificantly increases survival. Application of surgical treatment only is associated with a high rate of local recurrences. Chemoradiotherapy in penile cancer is significantly more effective vs each method alone. Radiation in a total focal dose more than 60 Gy improves local control over the tumor. Most effective are schemes of chemotherapy based on bleomycin. Polychemotherapy has no advantages over monochemotherapy with bleomycine. PMID- 15114749 TI - [An expanded technique of transrectal prostatic biopsy]. AB - To compare diagnostic value of transrectal prostatic biopsy in obtaining samples of tissue from different sites in patients with various levels of prostate specific antigen (PSA) and prostate size, we made primary transrectal biopsy of the prostate in 486 patients. The patients were divided into 7 groups by the number of punctures at biopsy (from 6 to 18). Among the patients with PSA under 20 ng/ml in the number of tissue biopsy samples 18, a rise in prostatic cancer detection rate (PCDR) was 16.6%. In PSA above 20 ng/ml, a statistically significant maximal rise in PCDR occurred in the increase of biopsy number from 6 to 12 (by 9.3%). The number of local cancer forms in patients with PSA < 20 ng/ml among all the detected cases rose from 70% (biopsy from 6 sites) to 92.3% (biopsy from 18 sites). Among the patients with PSA < 20 ng/ml and the size of the prostate > 50 sm3, a significant rise of PCDR increased from 20 to 33.3% in an increase of the puncture number from 6 to 12. In the group of patients with the same PSA level and prostate > 50 cm3 PCDR improves in biopsy from 14, 16 and 18 sites (from 12.1 to 27.7%, 28.5 to 33.3%, respectively). Standard biopsy is insufficient for adequate PCDR, it is necessary to obtain samples of tissue from a large number of sites with puncture of peripheral zone of the prostate. Transrectal biopsy of the prostate according to the extended method improves PCDR, primarily, in local cancer. PMID- 15114750 TI - [Extracorporeal shock-wave therapy in the treatment of Peyronie's disease]. AB - The authors analyse the results of treatment of 28 patients with Peyronie's disease using extracorporeal shock-wave lithotripsy (ESWL) performed on Dornier U15 lithotriptor. A total of 2-6 sessions were made, maximal number--12. The efficacy was controlled by clinical indices and ultrasonic investigation (Doppler mapping of the blood flow). ESWL proved to be efficient in the treatment of Peyronie's disease (PD), primarily, in patients with early disease before appearance of severe fibroplastic alterations. Less plaque vascularization by energetic Doppler mapping due to ESWL is an important diagnostic criterion of PD treatment efficacy. Conservative treatment is not indicated in marked deformities and plaque calcification, erectile dysfunction. Moreover, any injection into the tunica albuginea, especially complicated by hematomas may be a damaging factor which triggers fibrous inflammation. Such patients should be treated surgically. If the patient is interested in immediate results or is not interested in continuation of sexual life, the treatment is prognostically uneffective. Thus, ESWL is an effective, safe method of PD treatment but requires further study and accumulation of clinical experience. PMID- 15114751 TI - [Use of impase in the therapy of erectile dysfunction]. AB - Efficacy of impasse in therapy of sexual dysfunctions of vascular genesis was studied in 38 male patients aged 35-68 years. The drug was taken for 2 months. The control was conducted with ultrasonic dopplerography. Impasse showed high efficacy in therapy of vascular sexual dysfunctions especially in the group of patients aged 35-50 years with initial values of maximal systolic speed at least 15 cm/s. PMID- 15114752 TI - [Prolit in the treatment and prophylaxis of nephrolithiasis]. AB - In the course of prolit treatment there was a spontaneous evacuation of small concrements from renal calyces and ureters in 11 (27.5%) patients, dilution of the concrements and reduction in size of the latter were observed in prolit combination with blemaren (2 patients--5% and 7 patients--17.5%, respectively). Overall response was 50% while that to cyston--16.7%. Diuretic, anti-inflammatory and plastic actions of prolit manifested in 100% patients with disappearance of salts from urinary sediment, abatement of microhematuria and mild leukocyturia. Thus, prolit is a highly effective food additive in the treatment of nephrolithiasis and opens new perspectives in the treatment of urolithiasis. PMID- 15114753 TI - [Modification of the test pressure-flow for evaluation of detrusor contractility in absence of urination]. AB - The aim of the study was design and validation of the test for examination of detrusor contractility in the absence of urination. The method consists in registration of detrusor pressure in maximal speed of urinary flow in the course of artificial urination using Foley's catheter. Urethral resistance was modeled by raising drainage up to 40 cm above the level of the symphysis which was adjusted and found optimal in 48 of 179 examinees. The method allowed to select patients with infravesical obstruction (IVO) and normal detrusor contractility (n = 36) as well as patients with hypocontractility without IVO (n = 12). The test "pressure-flow" in artificial urination evaluates detrusor contractility in patients unable to urinate. The test distinquishes patients with normal detrusor contractility and those with different degree of its impairment. This is important in assessment and prognosis of deficient urination. PMID- 15114754 TI - [Retroperitoneoscopic operations in cystic lesions of the kidneys]. AB - In 1997-2002 retroperitoneoscopic dissection of renal cyst walls (RDRC) was made in 21 patients. Computed tomography, excretory urography, retrograde ureteropyelography were conducted in 14, 3 and 2 patients, respectively. Ultrasonic investigation was made in all the patients before and after operation. Dissection of the cystic walls was successful in 20 of 21 patients, material for cytological and histological examinations was also obtained. Advantages of endoscopic operations are now obvious, so RDRS is perspective in cystic lesions of the kidneys and its usage will increase. PMID- 15114755 TI - [Endoscopic nephropexy]. AB - In line with new prospects, introduction of laparoscopic technique in nephropexy gave rise to some problems. One of them--postoperative rotation of the kidney (RK). Upper-pole endovideosurgical nephropexy (EVSN) with prolen net performed in the diagnostic center "Zdoroviye" in 90 patients has demonstrated that late after surgery 9 (10%) patients had pains provoked by RK and abnormal renal circulation confirmed by Dopplerography. In 1998 specialists of the diagnostic center "Zdoroviye" developed a novel method of EVSN providing elimination of the ptosis and RK. A total of 88 patients have been operated since. 67 patients with nephroptosis and RK have undergone nephropexy with a splitted flap of the prolen net, 21 patients without RK--standard nephropexy. Long-term postoperative follow up showed that only 3 of 88 patients failed to retain normal velocity of the major renal blood flow, though they improved this velocity noticeably. Thus, a differentiated approach to EVSN based on dopplerographic examination of the major renal blood flow brings about improved treatment results in nephroptosis. PMID- 15114756 TI - [Gas-free lateral retroperitoneoscopy from an open mini-approach for adrenalectomy]. AB - The aim of the study was assessment of efficacy of gas-free lateral retroperitoneoscopy from the open mini-approach for adrenalectomy. Arguments are given for location of the mini-cut in the anterior segment of the 10th intercostal space and analysis was made of operative space parameters in retroperitoneoscopy from open mini-approach. The wound aperture 5.0 cm in size and 15.0 cm in depth is good as it provides the diameter of the monofocal zone 16.0 cm and bifocal zone 4.0 cm. If the size of the wound increases to 7.0 cm, the above parameters rise to 20.0 and 7.9 cm, respectively. Such visualized space is sufficient for adrenalectomy. Variants of the intervention are proposed depending on the side and kind of the lesion. A total of 21 patients with various adrenal pathology were operated. The operation lasted for 114.7 +/- 1.5 min. Overall blood loss reached 22.0 +/- 3.2 ml, transfusion--137.8 +/- 1.0 ml. The intestinal function normalized on day 1-2. In the postoperative period mean number of narcotic analgetics injections made up 4.9 +/- 1.5. Body temperature returned to normal on day 5.3 +/- 1.6 after the intervention. The length of the scar was 4.2 +/- 0.6 cm. None lethal cases were registered. Conversion to the classic open procedure had to be made in four cases. Gas-free lateral retroperitoneoscopic adrenalectomy from open mini-approach is minimally invasive operation. In well selected cases this is a safe and effective alternative to conventional and endovideoscopic gas surgery. PMID- 15114757 TI - [Pathogenetic mechanisms of infectious-inflammatory and thrombohemorrhagic complications of transurethral prostatic resection]. AB - The role of hemostatic disorders and alterations in urine proteolytic activity in pathogenesis of infectious-inflammatory and thrombohemorrhagic complications of prostatic transurethral resection (PTR) was studied. Hemostasis and urine proteolytic activity were investigated with biochemical and coagulological tests in 54 patients with benign prostatic hyperplasia (BPH) before and after PTR. Significant hypercoagulation in early postoperative period found in the examinees promoted formation of inflammation-related contaminated dense protein products which serve a source of permanent reinfection. The patients had low proteolytic activity of the urine caused by renal dysfunction as a result of abnormal urodynamics in this disease. Thus, hemostatic disorders and subnormal proteolytic activity of the urine are essential factors in pathogenesis of infectious inflammatory and thrombohemorrhagic complications of PTR. The conceptual scheme of postoperative complications pathogenesis in BPH patients subjected to PTR is proposed. PMID- 15114758 TI - [Surgical correction of primary nonrefluxing megaureter in children and its remote results]. AB - Correction of primary non-reflux megaureter (153 ureters) was made in 136 patients aged 3 months to 14 years. Bilateral disease was in 17 patients. Non reflux non-obstructive megaureter was in 113 cases, obstructive in 40 cases including association with ureterocele in 23 cases. Resection of distal ureter with its neoimplantation into the urinary bladder according to the antireflux technique was made in 146 patients, endovesical electroperforation and resection of ureterocele were made in 5 and 2 patients, respectively. Good results were obtained in 88.3% (135 ureters), satisfactory in 2.6% (4 ureters), unsatisfactory in 9.1% (14 ureters). After effective correction of megaureter, the treatment should be focused on adequate therapy of pyelonephritis present in 90% examinees, on improvement of urodynamics and stabilization of sclerotic process in renal parenchyma. The patients need long-term follow-up and more effective treatment. PMID- 15114759 TI - [Role of urodynamics correction in combined therapy of children with urinary infection]. AB - The authors propose differentiated complexes of therapy of children with urinary infection. These complexes include medicines (M-cholinolytics, antihypoxic drugs and drugs improving microcirculation) and low-invasive therapy. Clinical experience is reported for 4 boys and 40 girls with microbial-inflammatory urinary diseases. Standard urodynamic tests were made in all the patients before and after the course of therapy. Three variants of driptan effects on the urinary bladder were identified: M-cholinolytic, spasmolytic and mixed (larger volume and lower intravesical hypertension). Some of the patients experienced standard endoscopic reflux correction with the use of synthetic hydrophilic gel. Control urodynamic examinations 3 months after the treatment showed that functional disorders were reduced. Treatment efficacy was assessed with home flowmetry. Thus, combined therapy of microbial-inflammatory diseases with parallel correction of lower urinary tracts urodynamics by M-cholinolytics improves treatment results. Courses of antibacterial and uroceptic therapy should be followed by non-invasive monitoring to reveal urodynamic disorders, define dysfunction type and perform an additional treatment with mediators in combination with physiotherapeutic procedures. PMID- 15114760 TI - [A case of urethral transposition and vaginoplasty in the presence of urogenital sinus in women]. AB - A case of female false hermaphroditism is reported in a 30-year-old woman with urogenital sinus. This woman has undergone vaginoplasty and urethral transposition. The latter is not usually made in women with urogenital sinus. This leads to the position of the external urethral ostium in the depth of the newly formed vagina and, finally, to ectopia of the external urethral ostium or hypospadia. The result is that such woman suffers all life from urethritis, cystitis and vulvovaginitis. PMID- 15114761 TI - [A rare form of testicular teratoma]. PMID- 15114762 TI - [Renal resistance to ischemic damage and cell adaptation mechanisms]. PMID- 15114763 TI - [Memorable dates in the history of urology and allied fields in 2004]. PMID- 15114764 TI - Buruni ulcer disease. PMID- 15114765 TI - Progress towards measles elimination, western hemisphere, 2002-2003. PMID- 15114766 TI - [Clinical and pathophysiological aspects of the effects of chronic hypoxia/ischemia on energetic metabolism of the myocardium]. AB - Current studies are reviewed on pathogenesis of disorders in myocardial energy metabolism due to chronic long-term hypoxia of different etiology and myocardial ischemia with participation of some endogenic regulating factors including endocrine system. Literature data are presented on research into features of energy metabolism in stunned and hibernating myocardium in patients with ischemic heart disease. PMID- 15114767 TI - [Specific features of pathogenesis and diagnosis of hemorrhagic stroke in young patients]. AB - The paper deals with hemorrhagic stroke (HS) pathogenesis and diagnosis in young people. Among cerebrovascular diseases in the young acute hemorrhagic strokes take noticeable place. Arterial hypertension, diabetes mellitus, smoking, alcoholism are among risk factors of subarachnoidal hemorrhage (SAH). Massive hemorrhages occur in the rupture of arterial aneurysms and arteriovenous malformations. HS in the young may be caused by blood diseases, i.e. leukemias, hemophilias, idiopathic thrombocytopenic purpura, coagulopathies; vasculitis in diffuse diseases of the connective tissue; non-inflammatory arteriopathies; drug addiction. Genetic predisposition to HS development is discussed with focus to such diseases as a family form of moya-moya disease, glucocorticoid-depressed hyperaldosteronism, elastic pseudoxanthoma, Marfan's syndrome, renal olycystosis, Sturge-Veber syndrome. It is recommended to use wider updated methods of neurovisualization (CT, MRT, angiography) in diagnosis of HS. The conclusion is made that HS diagnosis, especially in the young, needs a multidisciplinary approach with active participation of neurologist, neurosurgeon, therapist, endocrinologist, hematologist. PMID- 15114768 TI - [Pharmacotherapy of portal hypertension]. AB - A brief information is reviewed on potential mechanisms of portal hypertension pathogenesis. Main groups of medicines used for prevention and treatment of hemorrhages from varicose esophageal veins are described. PMID- 15114769 TI - [Characteristics and prevalence of various types of left ventricular myocardial dysfunction in patients with chronic cardiac failure]. AB - Prevalence of systolic and diastolic myocardial dysfunctions of the left ventricle (SDLV, DDLV) was studied in 223 patients with chronic cardiac failure (CCF) arising in the presence of arterial hypertension (AH), coronary heart disease (CHD) and their combination. The diagnosis was made in outpatient diagnostic center and included patients free of distinct clinical symptoms of CHD. DDLV was rather prevalent in a functional class (FC) I of CCF, in AH patients with SDLV FC III and CHD patients. Mean duration of cardiovascular diseases (CVD) before CCF onset with various types of left ventricular dysfunction was determined. Echocardiographic, including Dopplerographic, criteria of SDLV and DDLV were studied and their significance depending on FC and etiology of CCF was ascertained. The study elicited regularities of CCF formation in patients with various cardiovascular diseases, impact of the latter on myocardial dysfunction, echocardiographic features of CCF by systolic and diastolic type. PMID- 15114770 TI - [Remodeling in cardiac failure of functional class I due to arterial hypertension associated with ischemic heart disease]. AB - Hemodynamics and ventricular remodeling were studied echocardiographically in 192 men with heart failure (NYHA functional class I), arterial hypertension (AH) of stage I-III and clinical picture of ischemic heart disease (IHD). The latter presented in the patients with stable angina pectoris of FC I-II (SAP), unstable angina pectoris (UAP) without foci, paroxysmal atrial fibrillation, acute myocardial infarction (MI), postinfarction cardiosclerosis (PC) with SAP or UAP. The control group consisted of 41 healthy men. The patients had AH stage 1. The patients and healthy controls differed significantly by the size of the aorta, left atrium, thickness of the interventricular septum and posterior wall of the left ventricle. There was a significant left-ventucular hypertrophy in the groups with patients with MI, SAP and PC, UAP and PC (p < 0.001). In these groups the type of left ventricular remodeling was characterized as excentric type of left ventricular hypertrophy without its delatation. Normal left ventricular geometry was in healthy men, SAP, UAP, paroxysmal actual fibrillation. PMID- 15114771 TI - [Quantitative assessment of some risk factors of chronic cardiac failure and non invasive diagnosis of its early (preclinical) stages]. AB - Factors of risk of chronic circulatory insufficiency (CCI) were assessed in 263 hypertensive patients with and without ischemic heart diseases. CCI was diagnosed in 218 of these 263 patients. Early stages of CCI may be diagnosed in certain number of these factors. ECG findings in isolated arterial hypertension detected left ventricular dysfunction classified as chronic cardiac failure of functional class I (NYHA criteria). PMID- 15114772 TI - [Hyperventilation syndrome in bronchial asthma, essential hypertension and organic neurosis. Clinical picture and external respiration function]. AB - The external respiration function was studied in 100 patients with hyperventilation syndrome (HVS) divided into 3 groups: 40 patients with HVS and bronchial asthma (group 1) consisting of 15 males and 25 females (age median--45 years, 25 percentile--37 years, 75 percentile--53 years); 39 patients with HVS and essential hypertension (group 2) consisting of 8 males and 31 females (age median 49, 25 percentile--40 years, 75 percentile--57 years); 21 patients with HVS without concurrent somatic diseases of group 3 (7 males, 14 females, age median 45 years, 25 percentile--28 years, 75 percentile--45 years). It is shown that different disorders of pulmonary ventilation correspond to different clinical manifestations of HVS. Thus, in bronchial obstruction (group 1) HVS manifests with "weak respiration", in restrictive pulmonary disorders (group 2) HVS manifests as "heavy respiration", in high parameters of bronchial permeability (group 3)--"shallow respiration". PMID- 15114773 TI - [Mental status of patients with ulcer disease]. AB - Questionnaire survey has been performed among 100 patients with duodenal ulcer (DU). Character building of these patients when they were children and adolescents was wring in 80 +/- 4 cases (women) and in 88.6 +/- 3.2 cases (men). Most significant negative factors were undercare, hypercare and living in incomplete family. The Leongard's questionnaire distinguishes various accentuations with prevalence of cycloid, epileptoid, emotive and demonstrative; anxiety scale was low. The Gissen personality questionnaire "Ego" confirmed frequent occurrence among DU patients of emotive, hysteroid, epileptoid features with non-flexible type of reaction. By Luscher's test, anxiety was low or moderate. Overall prevalence of personality disorders totalled 29 +/- 4.5 cases, mostly in women. It was proved experimentally that psychosomatic disorders are realized in target organs. In DU these organs were gastrointestinal (36 +/- 5.7 cases). Similar data about involvement of the gastrointestinal tract (GIT) were obtained in stress reactions. However, "ulcer personality" was not documented. Support of preferable realization of psychogenic factors action in GIT was obtained with Gissen somatic questionnaire. Operating psychogenic factors and their dynamics were characterized; emotional deprivation was leading. Among psychopathological manifestations, asthenodepressive symptoms prevailed in 76 +/- 4.3 cases. High efficiency of gestalt-psychotherapy and coaxil in the treatment of DU in patients with psychopathological disorders of the neurotic level were found. PMID- 15114774 TI - [Comorbid gastric and duodenal changes in chronic hepatitis C in young patients]. AB - Characteristics of the gastroduodenal zone, frequency of gastric mucosa contamination with Helicobacter pylori (HP) were studied in young patients with chronic hepatitis C (CHC). Affections of the stomach and the duodenum were encountered in 76.3% of CHC patients. They run, as a rule, latently and in most cases (80%) are associated with HP. Concurrent diseases of the upper gastroduodenal tract seem to have only a minor impact on the course of HCV infection. Therefore, it is better to avoid eradication of HP in young patients with CHC. PMID- 15114775 TI - [Comparative efficiency of conventional myelosuppressive chemotherapy and therapy including interferon alpha in patients with chronic myeloid leukemia]. AB - We compared efficacy of conventional myelosuppressive chemotherapy with myelosan, hydrealitalir and interferon-alpha (Ifn-a) therapy in 52 patients with chronic myeloid leukemia (CML). The latter treatment (Ifn-a was given in different doses) was conducted in 23 patients. Significant differences (p = 0.007) were found in medians of survival of patients on conventional myelosuppressive chemotherapy (40 months) and on Ifn-a therapy (61 months). Toxicity of treatment with Ifn-a (domestic reaferon, in particular) was mild. Thus, Ifn-a treatment was more effective than conventional myelosuppressive therapy against CML in a chronic phase. PMID- 15114776 TI - [Current approaches to treatment of urinary bladder cancer]. AB - The study included 30 patients with surface cancer of the urinary bladder stage TA-T1 G1-3. As the first step of the treatment, all the patients were operated with removal of the tumor then the patients were randomized to postoperative intravesicular immunotherapy with ronkoleucine in single doses 500,000 IU (15 patients of group 1) or 1,000,000 IU (15 patients of group 2). It was found that group 2 patients had recurrences much less frequently (26.7 vs 66.7%, respectively). With higher degrees of differentiation of the tumor cells recurrences occurred more frequently in both groups. Group 2 patients developed recurrences significantly less frequently in G1 and G2 (22.2%). In G3 all the patients had recurrences. Intravesicular administration of ronkoleukine raised absolute number of CD3 and CD4 subpopulations during the treatment and after it as well as raised concentration of TNF. The levels of the latter in the urine rose after the end of each immunoprophylaxis course. Intravesicular use of ronkoleukine entailed no specific toxic reactions. Thus, intravesicular prophylactic immunotherapy of recurrent surface cancer of the urinary bladder with ronkoleukine in a single dose 1,000,000 IU is effective prevention in patients with high (G1) and moderate (G2) grade of tumor cell differentiation. The single dose 500,000 IU is uneffective. A rise in subpopulations CD3, CD4 and TNF cytokine in the urine evidences for systemic activation of the immunity. PMID- 15114777 TI - [Efficacy of combined administration of ursodeoxycholic acid and hepthral in the treatment of primary biliary cirrhosis]. AB - The efficacy of ursodeoxycholic acid (UDCA) combined with s-adenosylmethionine (SAMe, heptral) and UDCA alone was studied in the treatment of 19 patients (age 36-57 years) with primary biliary cirrhosis (PBC). UDCA was given in a dose 10 mg/kg/day, heptral--in a dose 800-1600 mg/day. It was found that UDCA + SAMe treatment results in earlier and longer remission. Normalization of biochemical indicators of cholestase (levels of serum bilirubin, cholesterol, total bile acids, alkaline phosphatase, 5-nucleotidase and gamma-glutamyl transpeptidase) was recorded in 33% patients, cholestase diminished in the rest patients. Heptral potentiates cytoprotective and anticholestatic actions of UDCA but does not essentially influence its immunomodular and antiapoptotic effects. PMID- 15114778 TI - [Survival of patients with infectious endocarditis in various treatment methods]. AB - The aim of the study was to compare survival of patients with infectious endocarditis (IE) by abuse of intravenous narcotic substances in conservative and combined treatment in acute IE phase. A total of 195 IE patients were examined. 121 of them was treated conservatively, 74 patients received a combined treatment including surgical one. A long-term prognosis was studied by the Kaplan-Meier method. Among drug addicts with IE, the number of survivors in the observation period in early operation in the active IE phase was significantly higher than in the conservative therapy. Long-term survival of drug addicts with IE in combined treatment was also higher: 1-2- and 3-year survival was 85.8, 76.3 and 61.0% against 64.8, 49.0 and 34.2% in conservative treatment, respectively. In non addicts the differences were weaker: long-term survival in combined treatment was higher than in conservative one only in 5-year survival (74.1 and 41.6%, respectively). PMID- 15114779 TI - [Dynamics of lipid peroxidation and antioxidant defense in response to dalteparin therapy in acute coronary syndrome]. AB - The aim of the study was examination of lipid peroxidation (LPO) and antioxidant system (AOS) in patients with acute coronary syndrome (ACS); evaluation of the effect of dalteparin therapy on LPO and ACS. Fifty one patients (mean age 57.5 +/ 1.5 years) were randomized into two groups: group 1 (n = 31) received low molecular heparin dalteparin, group 2 (n = 17) was given non-fractionated heparin. The course of the anticoagulant therapy lasted 8 days. LPO and antioxidant defense in plasm and blood red cells were investigated on day 1 and 9. ACS patients appeared to have elevated LPO products production both in plasm and blood red cells, low plasm antioxidant activity (AOA). Dalteparin therapy lowered content of malonic dialdehyde (MD) in red cells and plasm, AOA enhanced. Heparin activity was lower: MDA in plasm decreased in a lesser degree while MDA in red cells continued to rise in lowering AOA in plasm and inhibition of catalase. PMID- 15114781 TI - [2004 calendar of memorable dates in the history of medicine]. PMID- 15114780 TI - [Arterial hypertension in aged patients in practice of therapists of the Ivanovo region]. AB - The authors analyse current practice of management of elderly patients with arterial hypertension in medical institutions of the Ivanovo region to find out factors impeding introduction of modern approaches to treatment of the hypertensive elderly. Medical documents were analysed for aged 291 outpatients and 178 inpatients with arterial hypertension (AH). 100 physicians filled in questionnaires. The following data were obtained: 91% physicians are sure about age-specific standards of arterial pressure; 85% physicians make their decision on elderly hypertensive patients' treatment basing on diastolic pressure; the necessity of AH treatment in the elderly is supported by only 6% physicians; nonpharmacological treatment of AH in practiced in 9% elderly patients; 40, 39, 28 and 16% patients receive ACE inhibitors, calcium antagonists, beta-blockers, diuretics, respectively; the percentage of unjustified hospitalizations is high; target arterial pressure was reached only in 0.9% hypertensive aged patients. It is concluded that the impeding factors for updating AH treatment in the region studied are the following: lack of information, stereotypes in medical practice, uneffective organization of medical aid for aged patients with hypertension. PMID- 15114782 TI - [Desensitization to diphtheria, tetanus and pertussis vaccine]. AB - Immunization with DTP vaccine (diphtheria, tetanus and pertussis) is a part of the vaccination calendar offered in childhood. Adverse allergic reactions vary from minimal urticarial reactions to life-threatening anaphylaxis. In infancy these reactions usually interrupt the vaccination calendar, but immunization in these children should be done. At the University Children's Hospital of Belgrade, a group of 137 children with suspected allergic anaphylactic reaction to DTP, DT, TT and monopertussis vaccine was studied for the last six years. Skin (prick and intradermal) tests were performed with corresponding vaccine. If both tests were negative, the vaccine could be given as a single dose of 0.5 ml. If one of these tests were positive desensitization with vaccine could be done (according to the protocol described by Carey and Meltzer). In one group of 52 children three days before desensitization, premedication with antihistamines, was done, whereas in the other group of 52 children premedication was not done. Two (3.8%) children in a group of 52 children with premedication had a minor (local) reaction after vaccination and 50 children (96.2%) had no reaction after vaccination, whereas no children (0%) had systemic reaction after desensitization. PMID- 15114783 TI - [Effect of preoperative factors on survival in patients with ruptured aneurysms of the abdominal aorta]. AB - Between 1991-2001 total number of 1058 patients was operated at the Institute of Cardiovascular Diseases of Serbian Clinical Centre due to abdominal aortic aneurysm. Of this number, 288 patients underwent urgent surgical treatment because of ruptured abdominal aortic aneurysm. The aim of this retrospective study was to show results of the early outcome of the surgical treatment of patients with ruptured abdominal aortic aneurysm, and to define relevant preoperative factors that influenced their survival. There were 83% male and 17% female patients in the study, mean aged 67 years. Intrahospital mortality that included intraoperative and postoperative deaths was 53.7%. Therefore, 46.3% patients survived surgical treatment and were released from hospital. Intraoperative mortality was 13.5%. Statistics showed that the gender and the age did not have any influence on mortality of our patients, as well as their co morbid conditions (p > 0.05). Clinical parameters at admission in hospital such as state of consciousness, systolic blood pressure, cardiac arrest and diuresis significantly influenced the outcome of treatment, as well as laboratory findings such as levels of hematocrit, hemoglobin, white blood cells, urea and creatinin (p < 0.05; p < 0.01). Ruptured abdominal aortic aneurysm still remains one of the most dramatic surgical states with very high mortality reported. We assume that important preoperative factors that influence the outcome of surgical treatment can be defined, but there is no single parameter which can certainly predict the lethal outcome after surgery. Also, the presence of co morbid conditions does not significantly influence the outcome of treatment in these patients. Therefore, urgent operation should not be withheld in most of the patients with ruptured abdominal aortic aneurysm. PMID- 15114784 TI - [Autoantibodies characteristic for autoimmune hepatitis found in chronic hepatitis C]. AB - In the patients with chronic hepatitis C (HHC) there have been found different autoantibodies, some typical for autoimmune hepatitis (AIH) type 1, like: antinuclear antibodies (ANA) in 15-60%, anti-smooth muscle antibodies (ASMA) in 34-60%, or AIH type 2: anti liver/kidney microsomes 1 autoantibodies (LKM 1) in 0 6%. Also, antimitochondrial autoantibodies (AMA), which defines PBC, is discovered in 0.7-1.5% patients with HHC. The purpose of the research was to determine the titr of the serum autoantibodies, which characterise AIH in the patients with HHC and to compare demographical, clinical, biochemical, histological and immunological parameters in the groups of patients: with or without autoantibodies. In 50 patients with HHC are defined clinical (demographical, troubles) facts, laboratorical (the values of complete bilirubin, ALT), histological diagnosis of the liver tissue (PH), and virusological diagnosis (antibodies and antigens of the hepatitis C virus), immunological (autoantibodies, immunoglobulins). The analysis of the results show us that ANA and AMA haven't been present in the patients with HHC in considerable titr (> 1:80), while in 10% of the patients there have been titr of ASMA. In 2 patients were both titrs, ANA (1:40) and ASMA (1:40), as well, which considered important, so there is entirely 7 patients (14%) with detected ANA and ASMA in considerable titr. Autoantibodies, which are characteristic for AIH 2, have not been found in the patients with HHC. Considered the sex, between the group of patients with HHC and autoantibodies, and the group of patients with HHC without autoantibodies, there is no important difference, although the percent of male is a little bit bigger in the second group. Considering the growth, there were no important difference between the patients groups. The fact that appearance of autoantibodies has no correlation with the growth, we can explain with the fact that she most of patients with HHC are persons from 21 to 30 years old, and that the older are no so present. The troubles are present in about the half patients with HHC, and it isn't proved that the appearance of autoantibodies have the influence on this particular clinical parameter. The comparison of activities of the disease, the analysis of the ALT values and the bilirubin, we also didn't get considerable difference between these two groups of patients. Three patients who haven't autoantibodies, but they have the ALT values expended between 5 and 10 times, which we consider the high grade of the necrosis. This high values we can't notice in the patients with autoantibodies. The parameters of grade inflammatory reactions of the extent values of Ig, so as IgG category, also doesn't show we the important difference between the groups. The most patients have normal values both: Ig and IgG. Despite that in both categories of the patients, there is no considerable difference in PH, as well as, in the stage of fibrosis, two patients without autoantibodies have more visible degree of necrosis, manifested like reasonable necrosis, while all the others patients have gently visible necrosis. It is important to say that the patients with autoantibodies have no histological indications for AIH, in the PH sense, and the presence of more important plasmatic infiltrate. All the patients with HHC and autoantibodies have higher values CIC, but the frequency of appearance is not different between the both groups, which show us the variety of becaming autoimmune and immune phenomena in HHC. Based on produced results, we can conclude that: 1. autoantibodies which are characteristic for AIH 1 are visible in 14% of the HHC patients, 2. there are more ASMA in HHC, then ANA; 3. there is no autoantibodies, which caracterize AIH 2, in HHC; 4. the growth and the sex of the patients with HHC have no influence on showing autoantibodies characteristic for AIH; 5. the presence of autoantibodies, characteristic for AIH 1, have no importance of showing the burdens in the patients, degree of PH and the stadium of the diseases, and also on the value of Ig and IgG; 6. discovering the autoantibodies in HHC, no matter there Is no considerable presence on it's symptomatology and the prognosis, oblige us on further following and analysis for starting antivirus therapy, In order to present eventual beginning of AIH. PMID- 15114785 TI - [Association of estrogen and progesterone receptor levels and the expected response to chemotherapy and hormone therapy in breast carcinoma with increased expression of human epidermal growth factor 2]. AB - The predictive value of Human Epidermal growth factor Receptor 2 (HER-2) on the response to chemotherapy and endocrine therapy in breast cancer patients has not yet been determined. The expression of other biomarkers in breast cancer can further influence the response to therapy. The aim of our study was to investigate if status of steroid receptors (SR) influenced the response to anthracycline-containing chemotherapy and tamoxifen in a group of HER-2 positive advanced breast cancer patients. Forty breast cancer patients, who were entered into the various prospective clinical trials conducted at the Institute of Oncology and Radiology of Serbia during their metastatic phase of disease, were involved into this analysis. Steroid receptors content were determined both by biochemical method and immunohistochemical (ICH) method, while HER-2 content were determined only by ICH method. Twelve out of 40 women were sequentially treated by anthracycline-containing chemotherapy and, always upon disease progression, with antiestrogen tamoxifen. The objective response to anthracycline therapy was obtained in 4 out of 12 patients (RR = 0.33, CI 95% = 0.05-0.61). In three of them the response to tamoxifen was noticed, as well. Of 8 anthracycline resistant patients in this group, 7 patients also had disease progression as best response to tamoxifen despite the fact that most of them (5 out of 7 tamoxifen resistant women) had positive SR status. Our results showed a trend (Fisher test, p = 0.06) that clinical response to anthracycline-containing chemotherapy might be of some predictive value for the response to subsequent tamoxifen therapy in HER-2 positive advanced breast cancer patients. However, these results were obtained on a small number of patients, so further investigation is warranted. PMID- 15114786 TI - [Effect of donor-specific blood transfusion on the outcome of kidney transplantation]. AB - Donor specific transfusion (DST) is proclaimed to improve graft survival in living related kidney transplantation (LRTx). The aim of the present study was to estimate the influence of DST on LRTx graft function, acute rejection rate (AR) and survival in the early and late posttransplant period. Fifty-five LRTx patients (grafted in the same year, and matched for recipients' and donor's age, sex) were included into the study. Ninety pts received DST: 4 patients were excluded from further evaluation (3 developed positive cross match reaction and one patients received cadaver graft) and 15 patients subsequently underwent LRTx from their respective blood donors (group 1). Their outcome was compared with 15 patients who had never been transfused before (group 2) and 25 random transfused patients (group 3). Besides similar patients' and donors' sex and age, kidney transplantations were performed in the same period. Graft functions were followed up 6-60 months after LRTx. DST protocol consists of 3 x 150 ml potentially related donor's fresh whole blood at 2-week intervals (DST1, DST2, DST3) with 3 days azathioprine administration (2 mg/kg bw, one, day before to one day after DST administration). Donor specific cytotoxic antibodies were determined before DST1, at the day of DST2, DST3 and 14 and 28 days after DST3. All patients were grafted at least one month after the DST3. Immunosuppressive protocol consisted of three drugs. There is no difference in HLA mismatches, MLC answer, and pretransplant panel reactive antibodies level between groups. One patient from group 2 lost their graft in the first postTx month (acute tubular necrosis). A better graft function was preserved in patients from groups 1 and 3 than group 2 in the observed periods. Number of patients with acute rejection was unsignificantly different: 5/15 from group 1, 12/25 from group 3 but 8/10 patients from group 2. However, the acute rejection rate was lower in patients from group 1. One and five-year graft survival was 100% for grafts from groups 1 and 3, while it is gradually decreased for group 2 grafts: 84.5% and 57%. Our results confirmed the beneficial effect of blood transfusion on LRTx renal graft function and survival and DST on the incidence of acute rejection. PMID- 15114787 TI - [First case of Malassezia globosa isolation in Serbia]. AB - Today is known that genus Malassezia includes seven species: M. furfur, M. sympodialis, M. obtusa, M. globosa, M. restricta, M. sloofflae and M. pachydermatis, but role of each of the species in the pathogenesis of disease has not been elucidated yet, so further laboratory isolation and identification are necessary. We report the first case of isolation of Malassezia globosa in Serbia (Belgrade), in a patient suffering from Pityriasis versicolor. Identification of M. globosa was based on macroscopic, microscopic and biochemical characteristics. Isolation was done on Leeming and Notman medium and on mDixona agar, at 350C, during 7 days in aerobic conditions. Also the yeast's biochemical phenotype was determined as catalase (+), lipase (+), esculin degradation (-), Tween (20, 40, 60 and 80) assimilation (-). M. globosa is a lipophilic yeast of the genus Malassezia and the common member of the skin flora. In concordance with some predisponing factors M. globosa is implicated in the pathogenesis of several skin diseases (pityriasis versicolor, malassezia foliculitis, seborheic dermatitis and some forms of atopic dermatitis). In immunocompromised patients and neonates this yeast can even cause fatal systemic infections. Because the role of Malassezia spp. In pathogenesis of skin disease is not still determined, we suggest laboratory diagnosis and identification of these species as a routine diagnostic procedure. PMID- 15114788 TI - [Burkitt-like lymphoma: subileus and ascites as the main clinical manifestations]. AB - Nodal presentation of Burkitt-like lymphoma is common, particularly in gastrointestinal tract. However, only few cases with massive ascites and signs of subileus due to lymphoma proliferation are described. We report a 31-year-old male patient who presented with fever, night sweats, vomiting and abdominal fullness. Physical examination suggested much ascites. Abdominal X-rays showed hydroaeric levels. Diagnosis or Burkitt-like lymphoma was established on the basis of cytological and immunohistochemical examination of ascites (immune phenotype of malignant cells was EMA-, NSE-, LCA+, CD10 -/+, CD20 +, IgM +, Ki-67 + 100%). After treatment with BMF protocol complete remission was achieved and retained for 2.5 years. Authors stressed that immunohistochemical examination of ascites has been proved as simple and efficient method for establishing precise diagnosis. In this way, laparotomy was avoided, which otherwise would be necessary due to exclusive abdominal localization of the disease. PMID- 15114789 TI - [Posterior reversible encephalopathy syndrome]. AB - INTRODUCTION: Reversible Posterior Leukoencephalopathy Syndrome was introduced into clinical practice in 1996 in order to describe unique syndrome, clinically expressed during hypertensive and uremic encephalopathy, eclampsia and during immunosuppressive therapy [1]. First clinical investigations showed that leucoencephalopathy is major characteristic of the syndrome, but further investigations showed no significant destruction in white cerebral tissue [2, 3, 4]. In majority of cases changes are localise in posterior irrigation area of the brain and in the most severe cases anterior region is also involved. Taking into consideration all above mentioned facts, the suggested term was Posterior Reversible Encephalopathy Syndrome (PRES) for the syndrome clinically expressed by neurological manifestations derived from cortical and subcortical changes localised in posterior regions of cerebral hemispheres, cerebral trunk and cerebellum [5]. CASE REPORT: Patient, aged 53 years, was re-hospitalized in Cardiovascular Institute "Dediwe" two months after successful aorto-coronary bypass performed in June 2001 due to the chest bone infection. During the treatment of the infection (according to the antibiogram) in September 2001, patient in evening hours developed headache and blurred vision. The recorded blood pressure was 210/120 mmHg so antihypertensive treatment was applied (Nifedipin and Furosemid). After this therapy there was no improvement and intensive headache with fatigue and loss of vision developed. Neurological examination revealed cortical blindness and left hemiparesis. Manitol (20%, 60 ccm every 3 hours) and i.v. Nytroglicerin (high blood pressure). Brain CT revealed oedema of parieto-occipital regions of both hemispheres, more emphasized on the right. (Figure 1a, b, c). There was no sign of focal ischemia even in deeper sections (Figure 1d, e, f). Following three days enormous high blood pressure values were registered. On the fourth day the significant clinical improvement occurred with lowering of blood pressure, better mental state and better vision. There was no sign of left hemiparesis on the 7th day. On the 9th day there were no symptoms or sign of disease. Control brain CT (15th day) was normal. ETHIOPATHOGENESIS: Most common causes of PRES are hypertensive encephalopathy [6-8], pre-eclampsia/eclampsia [9-12] cyclosporin A administration [13-22] and uremic encephalopathy [23]. There are several theories about the mechanism for PRES in hypertensive encephalopathy (reversible vasospasm and hyperperfusion) and administration of cyclosporin A (neurotoxic effect). CLINICAL PICTURE: Most common symptoms are headache, nausea, vomiting, confusion, behavioural changes, changes of conciousness (from somnolencia to stupor), vision disturbances (blurred vision, haemianopsia, cortical blindness) and epileptic manifestations (mostly focal attacks with secondary generalisation). Mental functions are characterised with decreased activity and reactivity, confusion, loss of concentration and mild type of amnesia. Lethargy is often initial sign, sometimes accompanied with phases of agitation. Stupor and coma rarely occurred. DIAGNOSIS: In patients with hypertensive encephalopathy and eclampsia high blod pressure is registered. Neurological examination revealed vision changes and damages of mental function as well as increased reflex activity. Today, brain MRI and CT are considered the most important diagnostic method for the diagnosis and follow-up of patients with PRES [6]. Brain MRI better detects smaller focal parenhim abnormalities than brain CT. The most often neuroradiological finding is relatively symmetrical oedema of white cerebral tissue in parieto-occipital regions of both cerebral hemispheres. Gray cerebral tissue is sometimes involved, usually in mild form of disease. Diagnosis of this "cortical" form of PRES is possible by MR FLAIR (Fluid-Attenuated Inversion Recovery) technique [5]. TREATMENT: Therapeutic strategy depends on the cause of PRES and clinical picture. Most important are blood pressure regulation (labetalol, nitroprusid, diuretici), control of epileptic attacks (phenytoin), anti-oedema therapy. (Manitol), Induction of vaginal delivery in eclampsia and discontinuation of cyclosporin therapy. In most cases there are no neurological manifestations after the 7th day, but some studies showed normalisation of clinical finding after one year and more. PMID- 15114790 TI - [Subclinical thyroid disease--should we treat, should we screen for it?]. AB - Subclinical thyroid disease is defined by an abnormally high (subclinical hypothyroidism) or low (subclinical hyperthyroidism) serum thyrotropin (TSH) with peripheral thyroid hormone concentrations within the laboratory reference ranges. Such abnormalities in thyroid function tests are very common in the population and have been extensively dealt with in textbooks and reviews. Subclinical hypothyroidism is common especially in elderly women. There is no clear evidence to date that subclinical hypothyroidism causes clinical hearth disease. However, mild thyroid gland failure, evidenced solely by elevation of the serum TSH concentration, may be associated with increased morbidity, particularly for cardiovascular disease and subtly decreased myocardial contractility. In subclinical hypothyroidism both cardiac structures and function remain normal at rest, but impaired ventricular function as well as cardiovascular and respiratory adaptation to effort may became unmasked during exercise. These changes are reversible when euthyroidism is restored. Subclinical hypothyroidism does result in small increase in low density lipoprotein cholesterol and a decrease in high density lipoprotein, changes that enhance the risk for development of atherossclerosis and coronary artery disease. Because undetected subclinical hypothyroidism during pregnancy may adversely affect the neuropsychological development and survival of the fetus and be associated with hypertension and toxemia, screening pregnant women has been advocated. In addition, data suggesting that subclinical hypothyroidism is associated with ovulatory dysfunction and infertility may make screening worthwhile in this population as well. The combination of an undetectable serum thyrotropin concentration, as measured by an assay with a threshold of detection that is 0.1 mU per liter or less, and normal serum triiodothyronine and thyroxine concentrations (usually at the upper end of the normal range) is known as subclinical hyperthyroidism. This condition reflects the facts that before clinical features of thyrotoxicosis are apparent, the thyrotrophs usually respond to minor increments in thyroid hormone concentrations, which remain within the normal range, by switching off the production and secretion of thyrotropin. In the absence of clinical signs of thyroid disease, and even after additional investigations such as isotope uptake and imaging and measurement of the thyrotropin receptor antibody concentration, it may be difficult to decide whether the pattern seen on thyroid function tests is a consequence of nonthyroidal illness and concomitant medication, underlyling thyroid autonomous function or the initial phase of thyroiditis. Routine screening for thyroid disease with thyroid function tests is not recommended for asymptomatic children or adults. This recommendation does not mean that clinicians should not monitor thyroid function in patients with a previous history of thyroid disease. There is insufficient evidence to recommend for or against screening for thyroid disease with thyroid function tests in high-risk patients, including elderly persons, postpartum women, and persons with Down syndrome, but recommendations may be made on other grounds, such as the higher prevalence of disease and the increased likelihood that symptoms of thyroid disease will be overlooked in these patients. If screening is performed, the preferred test is measurement of thyroid-stimulating hormone (TSH) using a sensitive immunometric or similar assay, because of its superior sensitivity and specificity. PMID- 15114791 TI - [Rosacea as a multisystemic disease]. AB - Rosacea is a chronic skin disorder, affecting the face and chest, and develops mostly in the 3rd to 6th decades of life. It is characterized by erythema, telangiectasias, and recurrent flushings. During the time of this chronic inflammation, skin typically develops papules, pustules, and swelling. Ocular involvement occurs in 3 to 58% of patients with skin changes. Common ocular signs include blepharoconjunctivitis, meibomitis, and dry eyes. Rosacea keratitis, when present, however, has a poor prognosis and may lead to blindness. Among skin diseases, Helicobacter pylori infection has been often related with rosacea. A higher prevalence of indigestion and Helicobacter pylori infection in rosacea patients than in healthy controls has been reported. However, no causal relation has been identified. On the other hand, oral treatment with metronidazole is beneficial in all of three mentioned manifestations of rosacea (skin, eye, indigestion). There is obvious need for multidisciplinary approach, and investigation to rosacea. PMID- 15114792 TI - [Evaluation of injury severity and autopsy verification of the clinical diagnosis]. PMID- 15114793 TI - [Dr. Vojislav J. Subbotic]. PMID- 15114794 TI - Acute upper-airway obstruction in a two-year-old child who ingested an herbicide preparation. PMID- 15114795 TI - Got milk? PMID- 15114796 TI - Looking back: some of the nervous and mental derangements of alcoholics in 1909. PMID- 15114797 TI - New directions in peritoneal dialysis patient training. AB - OBJECTIVE: To study the effect of training methods on selected patient outcomes in peritoneal dialysis patients. DESIGN: Multi-center, longitudinal prospective quasi-experimental design study conducted over a 2-year period. SETTING: Thirty two Gambro Healthcare peritoneal dialysis (PD) home training programs in the United States. SUBJECTS: New patients starting PD were trained on PD technique and diet using either an adult learning theory-based curriculum in the experimental group (PG) or non-standardized conventional training programs in the control group (CG). Excluded were patients who were non-English speaking, legally blind without sighted caregiver, nursing home residents, and those with previous exposure to PD training. METHODS: Information was collected by means of manual data collection tools and though the use of Gambro Healthcare computer system and was analyzed for statistical significance by Gambro Healthcare biostatistician. RESULTS: Compared with the CG, initial training took longer in the PG (PG = 29 hrs; CG = 22.6 hrs; p < .0001), and time required for retraining was less but not statistically significant (PG = 8.7 hrs; CG = 12.5 hrs; p = .1324). The peritonitis rate was less in the PG (28.2 per 1000 patient months) than in the CG (36.7 per 1000 patient months), but did not achieve statistical significance (p = .09783). Exit site infections (ESIs) were less in the PG than the CG (PG = 18.5; CG = 31.8; p = .00349). Dropout from PD to hemodialysis secondary to infection was less in the PG (1.6%) than in the CG (5.6%) (p = .0069). Measured on a scale with 4 being the best score, mean fluid balance scores in the PG were 3.41 compared to 3.25 in the CG (p < .0001), and mean compliance scores for the PG versus the CG were 3.62 and 3.52, respectively (p < .0001). Laboratory parameters between the two groups were significantly different only for Kt/V (PG = 2.4; CG = 2.3; p = 0.0107). CONCLUSION: Use of the adult learning theory-based training method curriculum was positively associated with improved patient outcomes in the PD population studied. PMID- 15114798 TI - Obstructive uropathy: considerations for the nephrology nurse. AB - Obstructive uropathy is the cause of renal failure in 16.2% of pediatric patients who undergo renal transplantation, 12.9% of those on dialysis, and 23.1% of those with chronic renal insufficiency, according to the annual report of the North American Pediatric Renal Transplant Cooperative Study (NAPRTCS). Successfully caring for patients with obstructive uropathy requires an understanding of the various congenital anomalies that create obstructive uropathy, the urological interventions used to treat them, management of the patient, and the nursing care required. PMID- 15114799 TI - Secondary hyperparathyroidism in chronic kidney disease: clinical consequences and challenges. AB - Secondary hyperparathyroidism, a characteristic manifestation of chronic kidney disease (CKD), arises from a series of abnormalities in the interrelated cascade that controls bone and mineral metabolism. Although bone disease is the most recognized consequence of secondary hyperparathyroidism (HPT), it also precipitates a series of potentially devastating effects on the cardiovascular, nervous, endocrine, and erythropoietic systems. Optimal management of secondary hyperparathyroidism should allow control of serum parathyroid hormone levels while preventing hyperphosphatemia, maintaining normal calcium levels, and providing adequate vitamin D supplementation. Current therapeutic approaches are frequently unsuccessful in achieving these goals and may, in fact, exacerbate risk factors that increase morbidity and mortality in the dialysis population. Development of new therapeutic approaches may provide improved control of secondary hyperparathyroidism in the future. PMID- 15114800 TI - The use of tissue plasminogen activator infusion to re-establish function of tunneled hemodialysis catheters. AB - PURPOSE: To evaluate the safety and efficacy of the recombinant tissue plasminogen activator alteplase in the clearance of poorly functioning tunneled hemodialysis catheters. METHODS: We retrospectively reviewed the outcomes of 25 patients who presented with poorly functioning hemodialysis catheters and were treated with alteplase. After confirming fluoroscopically the need for thrombolytic therapy, alteplase was administered over 2 hours as a 2.5 mg/hour/catheter lumen infusion (total 10 mg). Treatment was considered a clinical success if a flow rate of 250 mL or more per minute was established. RESULTS: Clinical success was achieved in each of 25 patients (100%). There were no thrombolytic-related complications. Catheter survival was extended 30 days in 54% of patients and 45 days in 33% of patients. CONCLUSION: Alteplase is a safe and effective means of producing clearance of blocked tunneled catheters. PMID- 15114801 TI - Renal regulation of acid-base balance. AB - Because maintaining a normal body pH is essential to the efficient functioning of many physiologic processes, the body has a number of mechanisms that prevent pH fluctuations. Some of these prevent minute-to-minute pH fluctuations over the course of the day, whereas others maintain pH balance from day to day. The kidney plays a key role in both processes. The renal process of bicarbonate reclamation prevents the loss of bicarbonate in the urine and, thus, maintains plasma levels of one substrate that is instrumental to preventing minute-to-minute pH fluctuations. The other renal process, bicarbonate regeneration, replenishes the body's supply of bicarbonate and, thus, maintains pH balance on a day-to-day basis. This article will discuss basic principles of acid-base physiology, the mechanisms that prevent fluctuations in body pH, and the renal processes involved in maintaining a homeostatic pH environment. PMID- 15114802 TI - Increased risk of anemia in dialysis patients with comorbid diseases. AB - Recent analyses conducted by the United States Renal Data System indicate that dialysis patients with concomitant comorbidities such as systemic lupus erythematosus, multiple myeloma, acquired immunodeficiency syndrome, or other secondary glomerulonephritis/vasculitis are at greater risk for anemia. Understanding the interrelationships between anemia and these comorbid diseases can help guide nephrology nurses in the development of individualized treatment plans that can ensure improved anemia-related outcomes in these populations. PMID- 15114803 TI - The role of physiotherapy in a hemodialysis unit. AB - This case study highlights the role of the physiotherapist and importance of providing multidisciplinary care for hemodialysis patients. Nurses are often in positions to identify patients who are in need of or could benefit from a physiotherapy consultation. In this case, the collaborative approach was highly successful in working with the patient to return to her previous pain-free state and her previous level of functioning. PMID- 15114804 TI - Development of an ABO verification policy for solid organ transplants. PMID- 15114805 TI - Patients must be free of the worry of paying for medications. PMID- 15114806 TI - Planning and fiscal responsibility will better assist patients with costs. PMID- 15114807 TI - Daily dialysis: nutritional implications and advantages for a state-of-the-art treatment option. PMID- 15114808 TI - Peritoneal dialysis and intraperitoneal insulin: how much? PMID- 15114809 TI - ANNA special interest groups--fast track groups that get the work done! AB - The SIGs have grown from informal meetings of members to discuss clinical practice issues in their sub-specialties to a formalized committee to address the needs of the association and the nephrology community. A SIG Statement of Purpose was developed in October 2003 as an effort to clearly articulate the complementary and collaborative function of the group in concert with the mission of ANNA: The purpose of the SIGs is to provide complementary and collaborative influence on the continuum of nephrology nursing practice through the promotion of advocacy, scholarship and excellence. With this purpose as a guide, the SIGs will continue to evolve as needs of the association and nephrology nursing change. PMID- 15114811 TI - It's in your hands.... PMID- 15114810 TI - Qualitative research. AB - A well thought out and conducted qualitative study provides a rich source of knowledge that can be used to provide new information about a phenomenon, promote insight and awareness of human experience and support the development of formal frameworks and tools (Kearney, 2001). At the most basic level, qualitative findings can encourage insight and empathy by raising nurses' awareness of "what it feels like to be in a given situation" (Kearney, 2001, p. 150). The study by Nagle (1998), for instance, draws nurses' attention to the disembodiment patients feel as they undergo hemodialysis and their struggle to develop a meaningful relationship with the technology they encounter on a daily basis. More formally, qualitative findings can also serve as the basis for assessment tools that allow nurses to compare the status of their patients with knowledge of a particular experience or illness trajectory (Kearney, 2001). For example, the grounded theory developed by Gregory et al. (1998) has served as the foundation for a quantitative measure of patients' perceptions of hemodialysis (Way, O'Brien, Gregory, Barrett, & Parfrey, 1999). This overview was intended to provide a general introduction to the main qualitative methods used in nursing research, and to briefly highlight some differences between qualitative and quantitative research. We hope that this review stimulates interest in qualitative approaches for nephrology nursing research. PMID- 15114812 TI - Casebook: chest infection in adults. PMID- 15114813 TI - The role of the GP in lung cancer. PMID- 15114814 TI - Diagnosing and managing sarcoidosis. PMID- 15114815 TI - The management of respiratory tuberculosis. PMID- 15114816 TI - Extrinsic allergic alveolitis in practice. PMID- 15114817 TI - Managing prostate cancer. PMID- 15114818 TI - Preparing for the oral module. AB - The MRCGP oral module is a rigorous assessment of the candidate's ability to make informed decisions about all aspects of general practice. Well-prepared candidates should have no problems in passing this examination. Familiarisation with the examination regulations and plenty of practice is essential. PMID- 15114820 TI - Is there a place for experimentation? PMID- 15114821 TI - How echographic image analysis of venous oedema reveals the benefits of leg elevation. AB - OBJECTIVE: Venous insufficiency, leading to venous oedema, is a key pathogenic factor for the non-healing of venous leg ulcers. This study aimed to evaluate the effects of leg elevation on venous oedema. METHOD: Ten patients aged 44-89 years (median: 61) with leg oedema had high-frequency B-mode ultrasound scanning and digital image analysis before and after three to four hours of leg elevation. The echographic image analysis system was used, where oedema is represented by the hypoechogenic part of the image--that is, the total number or density of low echogenic pixels (LEPs) in a particular area. RESULTS: Compared with pre elevation, the volume of the lower leg decreased by 2.9% +/- 0.6 (138 cm3 +/- 39) after three to four hours' elevation (p < 0.05). After elevation, the LEPs in the upper, middle and lower sites of the limb decreased by 8.8%, 15.6% and 17.3% respectively, reaching statistical significance (p < 0.05) in the lower site. The ratio of LEPs in the upper and lower dermis in the upper, middle and lower sites decreased by 30.3%, 45.8% and 22.5% respectively. This was significant in both the middle and lower sites (p < 0.01). After elevation dermal thickness increased by 0.047 mm, 0.194 mm and 0.232 mm respectively. This change was statistically significant in the middle (p < 0.05) and lower sites of the limb (p < 0.01). CONCLUSION: LEPs are a sensitive marker of dermal oedema and its effects. Leg elevation is extremely effective in reducing oedema, even if only for three to four hours. PMID- 15114822 TI - How safe is silver in wound care? AB - Use of silver in burns, chronic ulcers and diabetic ulcers can lead to circulatory absorption and deposits in wound sites, liver, kidney and other organs. Despite this, the risks of lasting tissue damage or functional disorders are low. PMID- 15114823 TI - An exploration of two opposing theories of wound contraction. AB - There are two schools of thought of how wound contraction occurs. Some researchers argue that myofibroblasts have contractile properties, whereas others propose that fibroblasts exert the force needed to achieve contraction. PMID- 15114824 TI - Combining topical negative pressure and a Bogota bag for managing a difficult laparostomy. AB - A patient with a severe intra-abdominal infection required a laparostomy. A Bogota bag was combined with topical negative pressure therapy, saving nursing time and enabling successful management of wound fluid. PMID- 15114825 TI - A silver sulphadiazine-impregnated lipidocolloid wound dressing to treat second degree burns. AB - OBJECTIVE: To evaluate the efficacy and tolerance of Urgotul SSD dressing (Laboratoires Urgo) in the treatment of second-degree burns. METHOD: This was a national multicentre phase III non-comparative open-label prospective study involving 10 burns units. The 41 subjects were non-immunosuppressed adults with second-degree thermal burn(s), which were clinically non-infected, less than 24 hours old, had a surface area less than 500 cm2 and warranted the local use of silver sulphadiazine. For four weeks, subjects were followed up weekly with a clinical assessment, bacteriological swabs and photographic recording. RESULTS: Of the 41 patients, 24 healed within a mean of 10.8 days and 13 had a skin graft on the study burn within a mean of 11.5 days. There were four premature study withdrawals. The total number of cumulative treatment days was 445, and 298 treatments were performed (including 257 dressing changes). Mean dressing wear time was 1.73 days. None of the subjects acquired a secondary infection. Researchers took 121 bacteriological samples, and wound colonisation with Staphylococcus aureus was found in only one patient. At follow-up nursing staff reported that dressing acceptability was good. CONCLUSION: Use of Urgotul SSD led to a good wound outcome--wounds healed or were grafted. PMID- 15114826 TI - Effect of glucose concentration on the growth of normal human dermal fibroblasts in vitro. AB - OBJECTIVE: Glucose requirements increase in tissue repair as glucose is an energy source for cell proliferation and the formation of extracellular matrix components. Glucose concentrations in leg ulcer wound fluid are lower than in normal human serum, with a median of 0.7 mM (range: 0.3-1.2 mM). This study investigated the effect of such low concentrations on the growth of fibroblasts in vitro. METHOD: Fibroblasts from 50-year-old and 87-year-old subjects were used. Growth in medium with various concentrations of glucose was determined by a colorimetric assay and microphotography. RESULTS: Up to day 6, there were minimal differences in growth, but after day 6 a clear dose-dependent increase in growth was observed. In the lower dose range (up to 2.3 mM), growth was highly dose dependent (r2 = 0.981), with an increase of 1.8 mM stimulating cell growth by day 10 up to 163% of the controls (p < 0.0001). Concentrations of 5.5-25.5 mM glucose stimulated cell growth by day 10 to about 210% of the controls (p < 0.0001), with little difference between these concentrations. CONCLUSION: These results suggest that glucose enhancement to cells involved in healing might serve as an adjunctive treatment for chronic wounds. PMID- 15114827 TI - Colloidal silver as an antimicrobial agent: fact or fiction? AB - OBJECTIVE: Colloidal silver preparations are marketed on the internet as omnipotent antimicrobial agents, but scientific support for these claims is lacking. This study reports the results of in vitro tests of colloidal silver's antimicrobial activity against several pathogenic or non-pathogenic microorganisms. METHOD: Three samples of colloidal silver were tested: one available commercially on the internet (silver concentration of 22 ppm) and two samples (concentrations of 403 and 413 ppm) which were prepared in our laboratory using standard chemical methods. RESULTS: In an agar-well diffusion assay none of the three colloidal silver solutions had any effect on the growth of the test organisms. All tested bacterial strains were sensitive to ciprofloxacin. Colloidal silver 22 ppm showed no bactericidal activity in phenol coefficient tests. CONCLUSION: As the tested colloidal silver solutions did not show any antimicrobial effect in vitro on the microorganisms, claims of colloidal silver's antimicrobial potency are misleading and there is no place for it as an antiseptic. PMID- 15114828 TI - How reliable is preliminary evidence? PMID- 15114829 TI - Dosage correction. PMID- 15114830 TI - Phytotherapy: an alternative treatment for non-healing ulcers. AB - There are few reports in the literature on the effectiveness of complimentary therapies on chronic wounds. Use of an aloe vera gel resulted in full healing after treatments such as antibiotics, surgical debridement and skin grafting had failed. PMID- 15114831 TI - [Diagnosis and treatment of insulinoma]. PMID- 15114832 TI - [Diagnosis and treatment for gastrinoma]. PMID- 15114833 TI - [Diagnosis and therapeutic strategies for non-functioning endocrine tumors of the pancreas]. PMID- 15114834 TI - [A patient with unresectable gastrointestinal stromal tumor responded to STI571]. PMID- 15114835 TI - [A case of pouchitis, duodenitis and pancreatitis showing diffuse irregular narrowing of main pancreatic duct after total colectomy for ulcerative colitis]. PMID- 15114836 TI - [A case of respiratory failure caused by pulmonary alveolar hemorrhage in Crohn's disease]. PMID- 15114837 TI - [A case of double cancer of squamous cell carcinoma and adenocarcinoma of the gallbladder]. PMID- 15114838 TI - [A case of hemorrhagic pancreatic pseudocyst presenting hemosuccus pancreaticus]. PMID- 15114839 TI - [Two-year survival and changes in the level of care for the elderly patients recognized as in need of long-term care in the public nursing-care insurance scheme]. AB - PURPOSE: The purpose of this study was to follow up survival and changes in the level of care needed for elderly patients 2 years (730 days) after recognition of their need for care in the public nursing-care insurance scheme, and to clarify the factors determining their prognosis in order to draw up intervention measures. METHODS: A total of 2,386 patients aged over 65 years (702 men and 1,684 women) who were recognized as in need of care between 21st October 1999 and 27th April 2000 were followed up with a multivariate logistic regression analysis. The dependent variable was life/death during the 2 years after recognition, and independent variables were sex, age, present address, the initial level of care needed, and care-requiring disorders. The same analysis was also applied to the 1,549 (440 mean and 1,109 women) who were still alive 2 years after recognition. The dependent variable was whether the level of care had deteriorated, and independent variables were the same as previously employed. RESULTS: When the odds ratio of survival was set at 1.0 for the male patients aged over 65 years, that for the females was 1.995. When the odds ratio was set at 1.0 for the patients aged 65-69 years, that for those aged 85 years and over was 0.593. When the odds ratio was set at 1.0 for the patients in need of long term support, that for those in level 2 was 0.458, that in level 3 was 0.326, that in level 4 was 0.257, and that in level 5 was 0.154, respectively. There were no significant differences in the odds ratios among the care-requiring disorders and present addresses. When the odds ratio for deterioration in the level of care was set at 1.0 for the patients aged 65-69 years who survived 2 years after recognition, that for those aged 80-84 years was 1.820, and that for those aged 85 years and over was 1.983. When the odds ratio was set at 1.0 for the patients with Alzheimer's disease, the values were 0.482 for dementia of unknown etiology, 0.110 for subarachnoid hemorrhage, 0.415 for cerebral hemorrhage, 0.431 for cerebral infarction, 0.240 for musculoskeletal diseases, and 0.359 for miscellaneous diseases. There were no significant differences in the odds ratios between sexes, among the initial levels of care, and among present addresses. CONCLUSION: The present 2 year cohort study of odds ratios of survival and deterioration in the level of care for the patients aged over 65 years revealed the survival of female patients to be twice that for males. The odds ratio of deterioration was higher for the patients aged 80-84 years and 85 years and over than those aged 65-69 years. The higher the initial level of care, the lower the survival odds ratio. The odds ratio of survival was lower for those aged 85 years and over than for those aged 65-69 years. These results indicate that the numbers of the elderly long-term care patients do not increase at the same rate as the numbers of the elderly. The odds ratio of deterioration was lower for patients with all diseases except vascular dementia than those with Alzheimer's disease. If reduction of the odds ratio of deterioration for the patients with all diseases except Alzheimer's disease and vascular dementia can be achieved by provision of sufficient services covered by public nursing-care insurance, such an approach should be stressed to prevent increase in the needs for long-term nursing-care. PMID- 15114840 TI - [The frequency of going outdoors, and physical, psychological and social functioning among community-dwelling older adults]. AB - PURPOSES: This cross-sectional study examined the relationship between the frequency of going outdoors and physical, psychological, and social functioning among community-dwelling older people. METHODS: Out of all residents aged 65 and over residing in Yoita town, Niigata prefecture (n = 1,673), 1,544 living at home participated in an interview survey held in November 2000. Physical, psychological, and social functioning were compared among four groups defined by the frequency of going outdoors: (1) at least once a day, (2) about once per 2-3 days, (3) about once a week, and (4) seldom. To examine the independent association between the frequency of going outdoors and potential factors, we used multiple logistic regression analysis. RESULTS: Overall distribution of the frequency of going outdoors among the subjects was 76.3% for at least once a day, 13.1% for about once per 2 or 3 days, 3.7% for about once a week, and 6.9% for seldom. The frequency of going outdoors did not differ between genders, but showed significant decrease with advancing age in both sexes. Elderly going outdoors more often were less functionally impaired, scored less for depression and were more socially active than their counterparts going outdoors less often. Multiple logistic regression analysis with the forced entry method identified walking difficulty and fear of falls as the most powerful independent factors associated with going outdoors less than or equal to once a week, while low social activity and a more depressed score were important factors associated with going outdoors about once per 2-3 days. CONCLUSIONS: The frequency of going outdoors may be regarded as a generic health indicator among community-dwelling elderly people. PMID- 15114841 TI - [Factors promoting follow-up services after medical examinations by municipal governments]. AB - OBJECTIVE: All Japanese municipal governments have a responsibility to conduct a medical examination for their residents aged 40 or more under the Health Services for the Elderly Act since 1984. According to the results of the medical examination, municipal governments should give appropriate follow up services to all users. The objective of this study was to find the factors promoting follow up services after medical examinations conducted by municipal governments. SUBJECTS AND METHODS: Mailed questionnaire surveys on follow up services after medical examination were sent out to all 3,255 municipal governments in Japan. The 2,447 municipal governments that answered all items on the questionnaire were enrolled in this study. Follow-up services by municipal government for persons with abnormal findings at medical examinations included "Suggestion to have a more detailed examination", "Confirmation of having had a detailed examination", "Individual health consultation", and "Home visit for medical guidance". These follow-up services were analysed in terms of the number of public health nurses per population and whether a continuous data set was maintained for medical examinations. RESULTS: Follow-up services were conducted more frequently in municipalities that maintained continuous data sets of medical examination than those that did not. They were also carried out more frequently in municipalities with higher numbers of health visitors per population than in those with lower numbers. The percentage of municipal governments that maintained continuous data of medical examinations was higher among those that conducted group medical examinations than those that conducted individual medical examinations at local medical facilities. CONCLUSION: Follow-up services after medical examinations by municipal governments showed a significant relationship with maintenance of a continuous data set of medical examinations and the number of public health nurses. PMID- 15114842 TI - [Several knowledge obtained through the application of PRECEDE-PROCEED Model on the health planning]. PMID- 15114843 TI - [Prevalence of exposure to environmental tobacco smoke among rural Japanese]. PMID- 15114844 TI - [Clinical analysis of idiopathic nonspecific interstitial pneumonia with unfavorable outcome]. AB - Nonspecific interstital pneumonia (NSIP) is associated with an excellent response to steroidal therapy. However, it was reported that some cases of NSIP might have an unfavorable outcome in spite of aggressive therapy. In this study, we explored the prognosis for 23 patients with NSIP and examined matched groups of 18 patients with treatment-responsive or naturally recovered idiopathic NSIP, and 5 patients with idiopathic NSIP who fared poorly despite therapy. The patients who had poor prognoses were, significantly, all non-smoking women. Histologic examination revealed that the cases with progressive, idiopathic NSIP were in all cases of the fibrotic type. By contrast, only 39% of those with a good prognosis, were fibrotic. No significant differences in CT appearance were noted between the two cohorts. These findings provide a potential clinical marker of therapy outcome of in idiopathic NSIP. PMID- 15114845 TI - [Grading of officially acknowledged respiratory disability and exercise disorders using cardiopulmonary exercise testing in patients with chronic respiratory diseases]. AB - This study examines whether the grading of officially acknowledged respiratory disability reflects exercise disorders in patients with chronic respiratory diseases. In order to do this, we analyzed the cardiopulmonary exercise testing (CPET) data of 258 patients with chronic obstructive pulmonary disease (COPD), 125 with sequela of pulmonary tuberculosis (TB) and 52 with interstitial pneumonia between 1989 and 2002. The peak oxygen uptakes of grade 1 and 3 COPD patients were 12.4 +/- 3.8 and 14.9 +/- 4.3 ml/min/kg (mean +/- SD), respectively. The worse the grade, the lower was the peak oxygen uptake, the differences being significant in COPD patients. Despite this, the peak oxygen uptake range overlapped greatly between grade 1 and grade 3 COPD patients. Sixty percent of patients with TB and 46% of patients with IP whose peak oxygen uptakes were similar to those of grade 1 COPD patients failed to be acknowledged as grade 1. There were no significant differences between the peak oxygen uptake of grade 1 COPD patients and grade 1 TB patients, grade 3 TB patients, and grade 4 IP patients. Patients with TB and IP showed more severe ventilatory and gas exchange disorders than those with COPD. We concluded that the present system of acknowledgement of respiratory disability in Japan did not accurately reflect exercise disorders in patients with chronic respiratory diseases. As well as this, we argue that there is discrimination between patients with COPD, TB and IP. It is necessary to establish an alternative system, reflecting exercise disorders evaluated by CPET to offer a more accurate acknowledgement. PMID- 15114846 TI - [Effects of smoking habits on pulmonary function. Cross-sectional and longitudinal studies in male subjects on medical check-up]. AB - Cross-sectional and longitudinal studies were done to evaluate effects of cigarette smoking and smoking cessation on age-related pulmonary function decline. Data on pulmonary function from 11,875 healthy asymptomatic men between the ages of 35 and 74 years were analyzed on the basis of smoking habits in each age group. Longitudinal changes in pulmonary function during a 5-yr period were also assessed in relation to smoking habits in 1888 healthy men. Cross-sectional studies showed that the difference of FEV1 between man current smokers and men who had never smoked is small at younger ages but increases with increases in age. A beneficial effect on FEV1 decline was observed in former smokers, even in less than 1 year after smoking cessation. Longitudinally, current smokers showed a more rapid decline in FEV1 in 5 years than nonsmokers. The men who quitted smoking had lower rates of decline in FEV1 than those who continued to smoke. These results indicate that cigarette smoking is associated with reduced pulmonary function and that smoking cessation may have a beneficial effect on FEV1 decline. Provision of a smoking cessation program for all smokers, especially those with a rapid decline of FEV1, should be considered a very important strategy to prevent progression of COPD. PMID- 15114847 TI - [Home respiratory care in remote areas and on small islands in Japan: status quo and problems]. AB - To investigate the status quo and problems regarding home respiratory care provided by medical institutes that played a central role in mountain villages and on small islands, we distributed a questionnaire in 250 remote medical institutes. The number of respondents was 116 (46.9%). Home oxygen therapy (HOT) and home mechanical ventilation (HMV) were carried out in 83 (71.6%) and seven (6%) institutes, respectively. The results of the survey regarding HOT in the remote medical institutes revealed that: (1) a large proportion of the patients were aged; (2) there were few cases per institute; (3) a high percentage of cases with chronic obstructive pulmonary disease (COPD) as the cause were evident; (4) visiting care was being provided in a high percentage of cases; and (5) few institutes were equipped with a spirometer. Overall examination of the responses showed that HOT was prevalent even in the remote medical institutes, and could be smoothly conducted, if communication with a medical institute specializing in respiratory diseases and an emergency transfer system could be established. On the other hand, the prevalence of HMV is still low, and problems such as the establishment of a medical team system and a caregiver support system has yet to be solved. PMID- 15114848 TI - [A case of farmer's lung disease manifested by smoking cessation]. AB - A 40-year-old man, a cattle hoof-chipper, was admitted to Hokkaido University Hospital in June 2002 for a workup of the occasional fever, cough and dyspnea that had affected him at work since March 2002. He had not smoked since January 2001. On admission, he had a marked increase in the proportion of lymphocytes (62.9%) and the CD 4/CD 8 ratio (3.0) in the bronchoalveolar lavage fluid. Serum precipitating antibody against Thermoactinomyces vulgaris was detected. The results of a transbronchial lung biopsy were unremarkable. Since his chest radiographs at the previous clinic showed small nodular opacities, we diagnosed farmer's lung on the basis of the diagnostic criteria defined by the Research Group of the Ministry of Health, Labor and Welfare. He was treated with prednisolone and advised to wear a protective mask, and his symptoms have not recurred since. This is an interesting case in which smoking cessation appears to have unmasked latent farmer's lung. PMID- 15114849 TI - [A case of MPO-ANCA positive vasculitis associated with diffuse alveolar hemorrhage and various cardiac conducting system abnormalities following propylthiouracil treatment]. AB - A 50-year-old woman had been treated with propylthiouracil (PTU) for hyperthyroidism. She was admitted to our hospital because of hemosputum, and severe hypoxemia developed. The CT scan showed diffuse infiltration in both lung fields, bronchoalveolar lavage fluid revealed diffuse alveolar hemorrhage, and the level of myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA) was high; and therefore diffuse alveolar hemorrhage associated with MPO-ANCA positive vasculitis induced by PTU was diagnosed. Following corticosteroid therapy initiated after the termination of PTU, the pulmonary infiltration rapidly improved and the patient's MPO-ANCA level returned to normal. Recrudescence of diffuse alveolar hemorrhage occurred following a reduction in steroids, but no recurrence was found after cyclophosphamide therapy was combined with steroid therapy. During the course of therapy, various cardiac conducting system abnormalities which correlate with the course of steroid therapy were found, indicating that cardiac conducting system abnormalities may be associated with MPO-ANCA-positive vasculitis. PMID- 15114850 TI - [Sjogren's syndrome with solitary nodular pulmonary amyloidosis]. AB - We describe a case of limited pulmonary amyloidosis with Sjogren syndrome. A 58 yr-old woman was referred to our hospital because of an abnormal chest radiograph (solitary small nodule) that was examined to investigate the cause of a persistent cough. A chest CT revealed a solitary small nodule in the left lower lung field. The specimens obtained by thoracoscopic surgery showed AL (kappa) amyloid deposits with lymphoplasmacytic infiltrate. Immunofixation of the serum and concentrated urine failed to demonstrate monoclonal immunoglobulins, and no amyloid deposits in the stomach were detected. She was subsequently diagnosed as having primary Sjogren's syndrome. Nodular pulmonary amyloidosis with Sjogren syndrome is very rare condition, and most cases present multiple nodules. As far as is known, this is the first report of a solitary nodule in pulmonary amyloidosis with Sjogren syndrome. PMID- 15114851 TI - [A case of a slow-growing, non-mucin-producing bronchioloalveolar carcinoma with a 9-year clinical history before thoracoscopic surgery]. AB - A 73-year-old woman was followed up with high-resolution computed tomography (HRCT) from 2000 through 2002 because an area of ground-glass opacity in area S1 + 2 of the left upper lobe was found on HRCT in August 2000. The opacity was present on ordinary CT scans obtained in 1993, but had not been recognized. Thoracoscopic partial resection of the left lung was performed on October 8, 2002. Pathologic examination of the resected specimen showed localized bronchioloalveolar carcinoma (Noguchi's type B). Tumor doubling time was 1718 days from 1993 to August 2000 and 273 days from August 2000 through 2002. This case of adenocarcinoma with bronchioloalveolar growth was characterized by initial slow growth followed by accelerated growth, no mucin production, and a long interval from the detection of opacity to surgery. PMID- 15114852 TI - [A case of vagus nerve schwannoma in the middle mediastinum presenting with dyspnea]. AB - A 67-year-old woman complaining of dyspnea was referred to our hospital because of an abnormal chest radiography shadow. Computed tomographic (CT) scans as well as magnetic resonance images (MRI) demonstrated a mid-mediastinal mass of 9 cm in diameter, surrounded by the superior vena cava (SVC), the ascending aorta, the azygos vein arch and the trachea at the subaortic level. The left main bronchus was stenotic, probably because of expansion of the tumor, which seemed to be the cause of the dyspnea. Routine examinations disclosed no metastatic or primary lesions elsewhere. The tumor was surgically resected. It measured 8 x 7 x 5 cm. The pathological diagnosis was benign vagus nerve Schwannoma, consisting of the histologies of both Antoni A and B types. We have reported this case because Schwannoma should be included in the differential diagnosis of mediastinal tumors even when it occurs in the middle of the mediastinum. PMID- 15114853 TI - [Chronic summer-type hypersensitivity pneumonitis presenting with acute exacerbation]. AB - A 77-year-old man was admitted to our hospital. He had first noticed a cough, sputum production, and low-grade fever during the summer of 1988. He was diagnosed as having summer-type hypersensitivity pneumonitis (HP) in 1989 on the basis of positive findings of anti-Trichosporon antibodies in the serum and bronchoalveolar lavage (BAL) fluid and and in environmental provocation tests. Thereafter, he presented with re-exacerbation of his disease in summer every year. In 1995, he retired from his work at a moldy hotel and moved to another house to avoid the causative antigen completely. There was no recurrence for the following 5 years. He suffered from cough, sputum production, and dyspnea 4 months after returning to his previous moldy house in October, 2000. A chest CT scan showed peribronchial and subpleural honeycombing, ground glass opacities, and traction bronchiectasis. BAL demonstrated that the lymphocyte count had changed from 78.9% to 42.9% and the CD4/CD8 ratio from 0.39 to 4.07 in 12 years. Antigen avoidance and steroid therapy have improved his condition. However, he stayed repeatedly in his moldy house, and finally died from acute exacerbation in March, 2002. Postmortem examination demonstrated diffuse alveolar damage (DAD) superimposed on honeycombing in the lungs. PMID- 15114854 TI - [Two cases of asthmatic exacerbation caused by parainfluenza virus 3 infection]. AB - Although viral respiratory tract infections are considered to be the most frequent causes of asthmatic exacerbation, respiratory viruses can rarely be detected in the adult population. We describe 2 cases, in a 43-yr-old man with severe atopic asthma and in a 69-yr-old man with moderate non-atopic asthma. After the onset of nasal discharge, sore throat and fever, the asthma had become exacerbated in both cases during the summer of 2002. In both cases, parainfluenza virus (PIV) 3 viral RNA could be detected from oral gargling by RT-PCR, and the serum viral antibody titer against PIV 3 increased significantly. These cases were therefore diagnosed as undergoing asthmatic exacerbation caused by PIV 3 infection and were successfully treated with systemic steroids. During summer, 2002, in our outpatient clinic, PIV 3 infection was demonstrated in approximately half of the asthmatic exacerbations associated with upper respiratory symptoms, including the present cases. Collectively, PIV 3 seems to represent an important viral cause of asthma exacerbation in summer. PMID- 15114855 TI - [A case of atypical pulmonary carcinoid accompanying skin metastasis]. AB - A 73-year-old woman underwent cranial surgery in 1999 after receiving a diagnosis of suspected malignant meningioma. She began complaining of headache 2 years postoperatively, and around the same time, she noticed a painful skin tumor. She was then transferred to our hospital for further evaluation. The skin tumor was diagnosed by skin biopsy as an atypical metastatic carcinoid tumor. Systemic examination demonstrated a primary lesion in the left lung. Pulmonary, skin and bone biopsy samples exhibited the same pathological findings as those of the atypical pulmonary carcinoid tumor. She did not show any carcinoid symptoms. EP therapy (etoposide + carboplatin) and CAV therapy (cyclophosphamide + doxorubicin + vincristin) were administered, but there was no clinical response. The patient is currently doing well without chemotherapy and is being followed by the Outpatient Department. PMID- 15114856 TI - [A case of spontaneous bilateral chylothorax]. AB - A 42-year-old woman sought medical attention because of a suddenly developed neck mass. Chest radiography disclosed bilateral pleural effusion, which turned out to be chylothorax. Although lymphatic leakage was suspected, no underlying cause was identified. Her condition resolved spontaneously in seven days. A few similar cases have been described in the literature, and are collectively referred to as "spontaneous bilateral chylothorax". PMID- 15114857 TI - [Pulmonary tuberculosis mimicking fungus ball]. AB - We treated a 42-year-old man with pulmonary tuberculosis presenting as a round mass in a cavitary lesion that resembled a fungus ball. These findings appeared within a short time. The diagnosis was confirmed by isolation of Mycobacterium tuberculosis from his sputum. He also had diabetes mellitus that was poorly controlled. The patient was treated with antituberculous chemotherapy and insulin therapy. With these treatments, the roentgenographic abnormalities resolved fairly rapidly. PMID- 15114858 TI - Mutational analysis of basic residues in the N-terminus of the rRNA:m6A methyltransferase ErmC'. AB - Erm methyltransferases mediate the resistance to the macrolide-lincosamide streptogramin B antibiotics via dimethylation of a specific adenine residue in 23S rRNA. The role of positively charged N-terminal residues of the ErmC' methyltransferase in RNA binding and/or catalysis was determined. Mutational analysis of amino acids K4 and K7 was performed and the mutants were characterized in in vivo and in vitro experiments. The K4 and K7 residues were suggested not to be essential for the enzyme activity but to provide a considerable support for the catalytic step of the reaction, probably by maintaining the optimum conformation of the transition state through interactions with the phosphate backbone of RNA. PMID- 15114859 TI - The effect of acetic acid, citric acid, and trisodium citrate in combination with different levels of water activity on the growth of Arcobacter butzleri in culture. AB - The influence of weak organic acids and trisodium citrate in combination with a high or a reduced water activity (aw) was investigated when a population of Arcobacter butzleri was exposed to a low concentration of acetic or citric acid, and trisodium citrate combined with high (0.993) and reduced (0.977) aw in culture broth at 30 degrees C. Regardless of water activity, acetic and citric acid (> 0.2%) inhibited the growth of A. butzleri with no viable cells detected after 4-5 h of incubation. Enhanced survival was found at reduced aw with addition of acetic acid. In contrast, after exposure to citric acid in combination with reduced aw inactivation was more rapid than that after being exposed to high water activity. Incorporation of trisodium citrate in combination with reduced aw (0.977) would probably not confer any extra protection. Concentrations of organic acid widely used in meat decontamination processing represent feasible tools for reducing A. butzleri contamination and hence the risk of Arcobacter infection. PMID- 15114860 TI - Purification and characterization of xylanases from Aspergillus giganteus. AB - A strain of Aspergillus giganteus cultivated in a medium with xylan produced two xylanases (xylanase I and II) which were purified to homogeneity. Their molar mass, estimated by SDS-PAGE, were 21 and 24 kDa, respectively. Both enzymes are glycoproteins with 50 degrees C temperature optimum; optimum pH was 6.0-6.5 for xylanase I and 6.0 for xylanase II. At 50 degrees C xylanase I exhibited higher thermostability than xylanase II. Hg2+, Cu2+ and SDS were strong inhibitors, 1,4 dithiothreitol stimulated the reaction of both enzymes. Both xylanases are xylan specific; kinetic parameters indicated higher efficiency in the hydrolysis of oat spelts xylan. In hydrolysis of this substrate, xylotriose, xylotetraose and larger xylooligosaccharides were released and hence the enzymes were classified as endoxylanases. PMID- 15114861 TI - Effect of glucose and light-dark environment on pigmentation profiles in the cyanobacterium Calothrix elenkenii. AB - Calothrix elenkenii was evaluated for accumulation of chlorophyll, beta-carotene and phycobiliproteins when grown under light-dark cycles and darkness, in the presence of glucose. Highest values of beta-carotene, chlorophyll, proteins and acetylene-reducing activity were recorded when the organism was grown in the presence of glucose under light-dark cycles after 15 d of incubation. The presence of glucose in the medium stimulated pigment production, especially of beta-carotene and chlorophyll, which may be due to increased energy-linked assimilation and ATP production. The photoheterotrophic potential of C. elenkenii can be exploited in value addition for increased and economic production of pigments. PMID- 15114862 TI - Characterization of chromate-sensitive and -tolerant mutants of Schizosaccharomyces pombe. AB - Stable chromium(VI)-sensitive and -tolerant mutants were obtained by induced mutagenesis of Schizosaccharomyces pombe lysine and leucine auxotrophic heterothallic strains 6chr+ and 9chr+. Eleven of them were selected for further studies. Fast transport of 51CrO4(2-) was detected in a representative sensitive mutant, chr-51S, while the tolerant mutant chr1-66T and the parental strain 6chr+ exhibited significantly lower 51CrO4(2-) uptake. The segregation of tetrads of three selected CrVI-tolerant mutants, chr1-66T, chr1-14T and chr2-04T, strongly indicated that tolerance was determined by single mutations. Random spore analysis proved that the mutations of chr1-66T and chr1-14T were allelic and the mutation of mutant chr2-04T was not allelic with the mutation of chr1-66T. Recombinants carrying the ura4D18 selective marker were created for transformation experiments. Two of them (chr1-661T and chr2-046T) can be used to clone and identify the genes responsible for their CrVI tolerance phenotype. PMID- 15114863 TI - Vegetative survival and reproduction under submerged and air-exposed conditions and vegetative survival as affected by salts, pesticides, and metals in aerial green alga Trentepohlia aurea. AB - Trentepohlia aurea vegetative cells do not survive submerged conditions for more than 5 months, but can survive air-exposed conditions for more than 1 year. Disintegration and rapid death of algal cells was observed to a higher extent under submerged than air-exposed condition. Under submerged conditions T. aurea did not form any sporangium while prolific formation occurred under air-exposed conditions. Under submerged conditions algal cells formed few-celled, filamentous, cytoplasmic type setae. Vegetative cells were resistant to some extent to various levels of salt (NaCl, < or = 0.8 mol/L), pesticides (DDT, 2,4-D or captan, 2000 ppm) and 'heavy' metals (zinc or nickel, 200 ppm; cobalt, < or = 100 ppm.) PMID- 15114864 TI - Isolation and selection of phenol-degrading microorganisms from industrial wastewaters and kinetics of the biodegradation. AB - Among 22 species of microorganisms isolated from phenol-containing wastewaters, Candida parapsilopsis was found to be capable of growth on a medium with 1 g/L phenol. Kinetic parameters of phenol biodegradation in a batch reactor were determined by measuring biomass growth rates and phenol concentration as a function of fermentation time. The Haldane equation described cell growth adequately, with kinetic constants mumax = 0.174/h, KS = 11.2 mg/L and Ki = 298 mg/L. PMID- 15114865 TI - Screening for pectinolytic activity of wood-rotting basidiomycetes and characterization of the enzymes. AB - Seventy-five fungal strains from different groups of basidiomycetes, newly isolated from rotten wood, were screened for pectinolytic activity. Despite the fact that basidiomycetes are scarcely referred to as pectinase producers, the polygalacturonase (PG) activity was detected in 76% of the strains; 16% with activity higher than 40 nkat/g, 40% between 13.3 and 40 nkat/g, and 44% with activity lower than 13.3 nkat/g. The highest productions were obtained among the fungi from order Aphyllophorales, family Polyporaceae. The characterization of the enzymes from the highest PG producers (Lentinus sp., Gloeophyllum striatum, Pycnoporus sanguineus, Schizophyllum commune) showed optimum temperature for catalytic activity at 60-70 degrees C and two peaks of pH optimum (3.5-4.5 and 8.5-9.5). The enzymes exhibited high pH stability (3.0-11.0) but after incubation at 40 degrees C for 1 h their activity dropped by 18-73%. PMID- 15114866 TI - Percentage of gelatinolytic bacteria among heterotrophic bacteria as indicator of water quality. AB - The relationship between the physiological group of gelatinolytic bacteria and the abundance of heterotrophic bacteria in freshwater ecosystems was described, based on analysis of 1082 different freshwater samples collected in Croatia. Percentages of gelatinolytic bacteria among the population of heterotrophic bacteria showed a significant negative correlation with the abundance of heterotrophic bacteria. The relation between the physiological group (gelatinolytic bacteria) and heterotrophic bacteria can be considered to be an indicator of the pollution degree of freshwaters. A high relative content of gelatinolytic bacteria (> 76%) always indicates the colony-forming units of heterotrophic bacteria < 1000/mL, which corresponds to the high water quality; gelatinolytic bacteria < 11% indicate polluted waters. Isolated strains of aerobically grown gelatinolytic bacteria were Gram-negative rod-shaped or Gram positive endospore-forming rod-shaped cells. PMID- 15114867 TI - Genotyping of fimbrial adhesins in Escherichia coli strains isolated from Slovak piglets suffering from diarrhea. AB - One-hundred sixty Escherichia coli isolates obtained from piglets with diarrhea from different parts of Slovakia were examined for the presence of genes coding for F4, F5, F6 and F41 fimbrial adhesins, and hemolytic activity. According to polymerase chain reaction tests 74 (46%) E. coli isolates were positive for primers that detected genes coding for fimbrial adhesins. Of these 74 isolates, 64 were positive for genes encoding for F4+, four for F5+, five for F6+, and one for both F41+ and F5+ adhesins. PMID- 15114868 TI - Effectiveness of Cyanothece spp. and Cyanospira capsulata exocellular polysaccharides as antiadhesive agents for blocking attachment of Helicobacter pylori to human gastric cells. AB - The effect of cyanobacterial polysaccharides (from Cyanothece spp. and Cyanospira capsulata) on the binding of Helicobacter pylori to gastric epithelial cells was evaluated. The antiadhesive action on Kato III and HeLa S3 human gastric cell lines was established. PMID- 15114869 TI - Double-disk synergy test positivity in Stenotrophomonas maltophilia clinical strains. AB - The double-disk synergy test (DDST) using Mueller-Hinton agar and antibiotic disks with centrally positioned disks of amoxicillin-clavulanate, ampicillin sulbactam, and piperacillin-tazobactam and, at a center-to-center distance of 25 30 mm, 2-4 disks with 10 various beta-lactam antibiotics per one plate was performed in 58 clinical isolates of Stenotrophomonas maltophilia to determine the effectivity of 3 beta-lactamase inhibitors. When tested with clavulanate as the central beta-lactamase inhibitor synergic action on tested strains was the most frequent with aztreonam (81.0% of strains), cefoperazone (63.8%), and cefepime (60.3%). With sulbactam the synergic action, i.e. DDST positivity, was high in the case of cefoperazone (15.5%), ampicillin, aztreonam and piperacillin (8.6% each); with tazobactam it was the most frequent with aztreonam (53.4%), cefoperazone (44.8%) and cefepime (37.9%). No synergy was demonstrated after application of meropenem regardless of the kind of beta-lactamase inhibitor used. In 58 strains of S. maltophilia, 55 different profiles of DDST positivity were found. The results confirm that clavulanate is the most effective inhibitor of S. maltophilia beta-lactamases. The utilization of DDST (performed in the recommended way) for the typization of strains Stenotrophomonas species and for the estimation of potential effectiveness combinations of beta-lactams with beta lactamase inhibitors for the therapy of stenotrophomonade infections was suggested. PMID- 15114870 TI - Determination of minimal regrowth concentration (MRC) in clinical isolates of various biofilm-forming bacteria. AB - Based on the ability to attach to polymeric surfaces, the formation of biofilms was determined in 5 wild-type strains (Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumanii, Escherichia coli, Staphylococcus warneri). Using modified Christensen method, minimum regrowth concentration (MRC) of piperacillin, piperacillin-tazobactam, cefoperazon, ceftazidim, cefepim, meronem, ciprofloxacin, netilmicin and amikacin for Gram-negative and of ampicillin sulbactam, chloramphenicol, tetracycline, clindamycin, vancomycin and teicoplanin for Gram-positive bacteria was estimated in trypticase-soy broth medium after a 1 d growth on polystyrene microtiter plates. Adherent bacterial populations exhibited reduced antimicrobial susceptibility, which was not shown in submerged cultures. Our results indicate that MRC can predict therapeutic outcome of antibiotic treatment better than the minimum inhibitory concentration tests commonly used. PMID- 15114871 TI - The detection of Chlamydia pneumoniae in aneurysm of abdominal aorta and in normal aortic wall of organ donors. AB - Sixty patients underwent surgery due to abdominal aortic aneurysms; the group included 30 patients with asymptomatic aneurysm and 30 with ruptured aneurysm. A control group comprised 30 organ donors. Surgical specimens derived from aneurysm or aorta fragments were investigated for Chlamydia pneumoniae DNA using PCR. In asymptomatic aneurysms, DNA was found in 9 cases (29%), and in ruptured aneurysms in 14 cases (49%). In the control group, C. pneumoniae DNA was not detected in an aortic wall. These results suggest that healthy aortic wall is not susceptible to chlamydial infection. A large number of aneurysm infections implies C. pneumoniae role in proteolysis and degradation of the aneurysm wall. The biological effect of this process may cause an enlargement of the aneurysm. PMID- 15114872 TI - Application of ARDRA and PLFA analysis in characterizing the bacterial communities of the food, gut and excrement of saprophagous larvae of Penthetria holosericea (Diptera: Bibionidae): a pilot study. AB - Amplified ribosomal DNA restriction analysis (ARDRA) was used to compare the bacterial communities of the food, the gut sections (ceca, anterior and posterior midgut, hindgut) and the excrement of the litter feeding bibionid larvae of Penthetria holosericea. For universal eubacterial primers ARDRA patterns were complex with only minor differences among samples. Taxon specific primers were also applied to characterize the samples. Fragment composition was transformed to presence/absence binary data and further analyzed. Cluster analysis revealed that bacterial communities of gut highly resembled each other with the exception of the ceca. ARDRA patterns of consumed leaves clustered together with the intact leaves but differed from those of the excrement. ARDRA results were compared with microbial community structure based on phospholipid fatty acid (PLFA) fingerprints. The cluster analysis of PLFA (presence/absence binary) data resulted in a pattern similar to the ARDRA data. The PCA analysis of PLFA relative content separated microbial communities into five groups: (1) anterior and posterior midgut, (2) hindgut, (3) ceca, (4) consumed and intact litter, (5) excrement. Both methods indicated that conditions in the larval gut result in formation of a specific microbial community which differs from both the food and excrement ones. Particularly ceca--(blind appendages, harbor very specific microbial community) are divided from the rest of the gut by perithropic membrane. PMID- 15114874 TI - [Changing one's point of view]. PMID- 15114873 TI - Chitosanase activity in Bacillus thuringiensis. AB - The ability to produce extracellular chitosanase (EC 3.2.1.132) was found by plate assays in 18 (23%) out of 77 crystalliferous strains of Bacillus thuringiensis. The best chitosanase producer was selected after the growth chosen in a liquid medium with colloidal chitosan as carbon source. Enzyme production was optimized (a 4-d incubation at 32 degrees C with shaking in a medium of pH 6.5 with 4% colloidal chitosan) and the enzyme was partially characterized. This is the first report on the chitosanase of B. thuringiensis. PMID- 15114875 TI - [Pain management--the role of the nurse]. PMID- 15114876 TI - [Middle back pain]. PMID- 15114877 TI - [Nursing competence in pain therapy]. PMID- 15114878 TI - [Wound nursing is more than management]. PMID- 15114879 TI - [Four nurses to see patients]. PMID- 15114880 TI - [Foreign nurses collaborate in the hospital]. PMID- 15114882 TI - [Sexual harassment in the workplace]. PMID- 15114881 TI - [We barely come too]. PMID- 15114883 TI - [Project: focus documentation should give consistent information]. PMID- 15114884 TI - The health hazards of smoking. PMID- 15114885 TI - Epidemiologic study of the association between human T-cell lymphotropic virus type 1 and Strongyloides stercoralis infection in female blood donors (Guadeloupe, French West Indies). AB - This case control study was conducted among female blood donors in Guadeloupe. A total of 85 HTLV-1 positive subjects were matched by age (+/- 5 years) in a 1:3 ratio to 255 HTLV-1 negative controls. The mean age was 48.5 and 48.9 years respectively. Ethnic, environmental and socio-economic risk factors were studied. Four risk factors were found independently associated with HTLV-1 seropositivity: Strongyloides stercoralis antibodies (OR = 3.6, CI 1.8, 7.3), low socio-economic status (OR = 2.0 CI 1.0, 4.0), low educational level (OR = 2.0 CI 1.0, 4.0) and agricultural activity (OR = 2.8 CI 1.4, 5.6). This logistic regression analysis demonstrates the reality of the association between HTLV-1 carriers and Strongyloides stercoralis, and its association with socio-economic and environmental risk factors. PMID- 15114886 TI - Features of isolated post-challenge hyperglycaemia in Jamaican adults. AB - Isolated post-challenge hyperglycaemia (IPH) can be defined as a two-hour plasma glucose concentration > or = 11.1 mmol/L with a fasting plasma glucose concentration < 7.0 mmol/L. The aim of this prospective study was to determine the prevalence of IPH in a cohort of Jamaican individuals, and to determine if simple clinical features may predict the presence and subsequent diagnosis of IPH. A cohort of 1694 adults aged 25-74 years without physician-diagnosed diabetes mellitus was randomly selected. An oral glucose tolerance test (OGTT) was performed. Anthropometry, blood pressure and lipid profiles were measured. The prevalence of undiagnosed diabetes mellitus by the 1999 World Health Organization criteria was 6.4%. IPH accounted for 24% of these cases and 1.4% of the entire population. Individuals with IPH were significantly older, with greater body mass index, waist-hip ratio, systolic blood pressure, fasting blood glucose, total cholesterol and LDL-cholesterol than individuals with normal glucose tolerance. Individuals with IPH were not significantly different from individuals with fasting plasma glucose levels > or = 7 mmol/L (i.e. fasting hyperglycaemia) in anthropometry or blood pressure. However, total cholesterol and LDL-C were significantly elevated in the IPH group. OGTT screening of individuals with impaired fasting glucose (i.e. 6.1-6.9 mmol/l) could reduce the IPH group by 50%. Reducing the threshold for fasting glucose to 5.6 mmol/L would correctly classify 87% of the population. We concluded that individuals with IPH have features of the metabolic syndrome, which can aid in selection for screening. OGTT screening of individuals with fasting glucose values of 5.6-6.9 mmol/l is needed to identify IPH. PMID- 15114887 TI - Excess healthcare cost associated with a low smoking prevalence, Barbados. AB - Policy makers need justification for smoking prevention. Barbados has debated but not enacted policies on tobacco control. This study estimated tobacco-associated morbidity and hospital care costs in order to justify prevention in a developing country with a low smoking prevalence of 9%. Hospital files of patients of index diseases and other chronic diseases with recorded status of smoking were followed on their outpatient, inpatient, elective and emergency attendance up to last discharge and analyzed as a nested case-control in a cohort of smokers and non smokers. Outcome measure was excess healthcare financial burden per person per year derived from morbidity incidence density, risk ratios, preventive fractions and excess hospital services consumed per person per year. Of 258 hospital patients, 84 were smokers. The mean follow up was 5.2 years. Eighty-four smokers had 463 excess outpatient appointments, 43 excess admissions, and 2651.6 excess hospital days. There were 44.8 fewer elective admissions and 62.24 more emergency admissions among smokers who suffered frequent and severer complications, (odds ratio = 3.78 for > or = 3 complications, p for Chi square trend, 0.0223), and had poorer prognosis translating into higher care costs, personnel effort, time, and human suffering. The excess hospital care cost was BDS$2,267 per smoker per year. Despite a low prevalence of smoking, tobacco caused a substantial public health burden. Hospital care cost for patients who smoked was 1.86 times higher than for non-smokers, and five times more than the government per capita health allocation. The annual excess hospital care costs in 1556 similar smokers would have exceeded the annual tobacco revenue. PMID- 15114888 TI - The emergence of Stenotrophomonas maltophilia as a significant nosocomial pathogen at the University Hospital of the West Indies. AB - Stenotrophomonas maltophilia is emerging worldwide as a nosocomial pathogen. It is associated with certain risk factors and a wide range of infections. This study was done to document its emergence at the University Hospital of the West Indies and to determine the incidence, distribution and risk factors associated with it. A retrospective study was conducted over the period April 1997 to December 2000. Clinical records were available for 46 of the 66 patients identified over the study period. Fifty-five per cent of the cases came from the Intensive Care Unit (ICU) and the rest from other wards. There was a slight increase in the prevalence of infection with increasing age. The surgical service accounted for the largest number of isolates. Of the cases presented, 95.7% were exposed to a wide range of antibiotics and had some form of instrumentation. Underlying disease was found in 71.7% of the patients. S maltophilia was found most often in the sputum of ICU patients whereas it was most often isolated from wound swabs in the ward patients. The organism was isolated from blood more often in ICU patients (23.3%) than in ward patients (9.5%) and there was a 44% mortality rate among the cases in ICU compared with those on the wards (4.8%). Stenotrophomonas maltophilia is an important nosocomial pathogen and occurs in a wide cross-section of patients. The risk factors must be addressed and infection control measures implemented to restrict the spread of this organism. PMID- 15114889 TI - Newborn circumcision using the Plastibell device: an audit of practice. AB - Circumcision of newborn male infants is widely practiced but controversial. Our experience gained circumcising 205 Jamaican neonates, using the Plastibell device is presented. Circumcisions, were requested by parents usually for hygienic reasons, and were brief outpatient procedures. Subcutaneous penile ring block with 1% lidocaine provided the most effective form of procedural analgesia. Bell separation usually occurred within 10 days of the procedure. Cosmetic results met with unanimous parental acceptance. Minor complications occurred in 2.4% of circumcisions. Physicians circumcising newborns must be readily accessible post procedure to address parental concerns and allay anxiety. PMID- 15114890 TI - A simple preparation for introductory training in the construction of distal coronary anastomoses. AB - A simple laboratory preparation for use in training of junior level residents in the construction of distal coronary anastomoses is described. The preparation is easily stored and rapidly set-up, and provides a realistic substrate upon which basic anastomotic technical skills can be taught and the trainee's effort can be immediately assessed. PMID- 15114891 TI - A simple, inexpensive simulation exercise in the construction of sutured intra thoracic oesophageal anastomoses. AB - The ability to perform a hand-sutured intra-thoracic oesophago-gastric anastomosis remains an important skill for surgeons in the developing world to master. In the developed world, this skill is no longer widely practiced due to the prevalent use of surgical staplers in most centres. A simple preparation for teaching aspects of hand-sutured anastomotic techniques is described. The set-up accurately evokes the spatial relationships and restrictions encountered during the course of an intra-thoracic oesophago-gastric anastomosis, and enables trainees to gain immediate feedback regarding their progress towards mastering this important thoracic surgical skill. PMID- 15114892 TI - Exploring knowledge and attitudes about emergency contraceptive pills among university students in Jamaica. A qualitative approach. AB - Emergency contraceptive pills (ECPs) are an important option for university students who may be at high risk for unplanned pregnancies. In the Caribbean, little research has been carried out on university student's knowledge and opinions of this method. This study uses qualitative methodology to explore knowledge and opinions on ECPs among university students attending The University of the West Indies (UWI) Mona campus. We conducted eight focus groups (n = 71) with female and male university students at The University of the West Indies, Kingston, Jamaica, in March 2000. The group discussions lasted approximately two hours. We tape-recorded discussions and then transcribed and analyzed them by coding responses according to themes. General knowledge of ECPs was high, but students lacked specific information about the method such as its time frame and its mechanism of action. Most students supported the method, especially after learning correct information. However, several students were concerned about its side effects and the potential for abuse or irresponsible use by young adults. Although the university students in this study lacked detailed information about ECPs, their opinion toward the method was favourable. We suggest further research to investigate the prevalence of misinformation about the method among other groups of Jamaicans. PMID- 15114893 TI - Effect of the oral contraceptive pill on patients undergoing controlled ovarian hyperstimulation. AB - This is a retrospective analysis of 89 patients who were undergoing controlled ovarian hyperstimulation for in vitro fertilization and embryo transfer in the Fertility Management Unit of the Department of Obstetrics, Gynaecology and Child Health, The University of the West Indies. Twenty-eight patients (Group A), who did not receive oral contraceptive pills prior to controlled ovarian hyperstimulation (COH) were compared with 61 patients in Group B treated with oral contraceptive pills for two months prior to undergoing COH assisted reproduction using the long protocol. The number of follicles, oocytes, estimated oestradiol levels on the day of administration of human chorionic gonadotrophin (hCG), pregnancy rates, miscarriage rates and the incidence of patients who developed ovarian hyperstimulation syndrome (OHSS) were the main outcome measures. The mean age and haematocrit were the same in each group. The number of follicles retrieved tended to be higher in Group A than in Group B (median 8 versus 6, p = 0.06) with significantly more oocytes being retrieved in Group A than Group B (p < 0.05). There were no statistically significant differences between the two groups in oestradiol levels, the proportion of patients with polycystic ovarian disease, the proportion of women who developed ovarian hyper stimulation syndrome or pregnancy outcomes. There was no difference between the groups in measures of clinical severity of OHSS. In a logistic regression model the significant predictors of OHSS were haematocrit and oestradiol levels. There appeared to be no significant clinical benefit in administering oral contraceptive pills for two months to patients prior to COH. PMID- 15114894 TI - Endoscopic management of post-cholecystectomy bile leak. AB - Endoscopic management of post cholecystectomy biliary leak is described in a 56 year-old patient who developed a cystic duct leak following open cholecystectomy. PMID- 15114895 TI - Thyrotoxic hypokalaemic paralysis in a pregnant Afro-Caribbean woman. A case report and review of the literature. AB - This paper reports the case of a 21-year-old Afro-Caribbean pregnant woman with hyperthyroidism and hypokalaemic quadriparesis and reviews the literature on the topic. Thyrotoxic periodic paralysis is a very rare condition in the Caribbean. This case reminds West Indian physicians to consider this rare condition in any patient that presents with paralysis. PMID- 15114896 TI - Acute poisoning in children. Cases hospitalized during a three-year period in Trinidad. AB - The aim of this study is to describe the epidemiological features of acute poisoning in children less than 16 years old who were admitted to a paediatric hospital in north Trinidad. The specific objectives included the determination of the age range most susceptible to poisoning, which agents are mainly responsible, an examination of the need for preventive strategies and educational programmes as well as to evaluate the need for a poison control centre in the country. Data were extracted from the medical records of 169 patients (83 males (49%) and 86 females (51%) with acute poisoning during the period of January 1998 to December 2000. The results revealed that the majority of cases of poisoning were accidental (84.6%), suicide (11.2%) and forced poisoning (4.1%). The largest category of poisoning was a miscellaneous group (24.8%) followed by the drug category (21.8%), kerosene (19.5%), pesticides (15.9%) and bleach (9.4%). Paraquat ingestion constituted 5.3% of cases. The highest prevalence of acute poisoning occurred within the age group of 0-4 years (69.2%), followed by the age group of 10-13 years (13.6%), 5-9 years (9.4%) and the age group with the lowest incidence was 14-16 years (7.6%). The only fatality was a female (10-13 year group) and this was due to suicidal ingestion of paraquat. All other cases were treated and subsequently discharged. The frequency of accidental poisoning in Trinidad merits more widespread public education aimed at preventing exposure to toxic substances while increasing the use of deterrents such as child-resistant containers. PMID- 15114897 TI - Non-ulcerative sexually transmitted disease and human immunodeficiency virus infection in patients attending a sexually transmitted disease clinic in Jamaica. PMID- 15114898 TI - High-grade sarcoma of the uterine corpus with heterologous elements and synchronous adenocarcinoma in an endocervical polyp. PMID- 15114899 TI - Failure to detect Angiostrongylus cantonensis in rats in Barbados. PMID- 15114900 TI - Cross-notation number priming investigated at different stimulus onset asynchronies in parity and naming tasks. AB - In this paper, a parity judgment task and a number naming task were used to investigate cross-notational number priming. Primes and targets could be verbal (e.g., seven) or Arabic numbers (e.g., 7), and were always presented in a different notation within the same trial (either a verbal prime and an Arabic target or an Arabic prime and a verbal target). Previous experiments showed that response latencies increase when the distance between prime and target increases (for example, in a naming task, seven is pronounced faster after 6 than after 5). This semantic distance priming effect was the same for Arabic and verbal targets and was the same for within-notation trials as for cross-notation trials. In the present experiments, we wanted to investigate whether the cross-notational priming effect also occurs at SOAs shorter than the ones used in previous experiments. Therefore, we used SOAs of 43, 57, 86, and 115 ms. Semantic distance effects were indeed present at these shorter SOAs: Processing times in the semantic parity judgment task and in the non-semantic naming task increased when the distance between prime and target increased. The results are discussed and integrated within an interactive dual-route model of number processing that postulates that the impact of the semantic and the non-semantic route depends on the task and the notation of the stimuli. PMID- 15114901 TI - Oculomotor bias induced by number perception. AB - Previous research with manual response methods has found evidence for an association between numbers and space. The present study investigated whether eye movements also show this association. Eye movement responses were recorded from 15 healthy participants as they categorized the digits 0-9 as odd or even. Responses were initiated faster to the left in response to small digits and faster to the right in response to large digits. Movement amplitudes were not systematically affected by either number magnitude or parity. These results provide further evidence for a spatially oriented "mental number line". PMID- 15114902 TI - Sequential modulations of logical-recoding operations in the Simon task. AB - The Simon effect consists of faster responses to the color (or another nonspatial feature) of spatially corresponding stimuli than to spatially noncorresponding stimuli. Recently, several studies observed the Simon effect after corresponding predecessor trials, but not after noncorresponding predecessor trials. To explain these sequential modulations, Sturmer et al. (2002) proposed a mechanism modulating the ability of stimulus position to automatically activate a response. The present study investigated which events are effectively triggering this mechanism in a variant of the Simon task, in which both stimuli and responses varied in color (participants wore colored gloves) as well as in horizontal position. In the same-color task (e.g., green stimulus-green response), a normal Simon effect showed up after corresponding trials, but no effect occurred after noncorresponding trials. In the alternate-color task (e.g., green stimulus-red hand), no effect occurred after spatially corresponding trials, whereas an inverted Simon effect was found after noncorresponding trials. Additional analyses showed that repetition (or alternation) effects did not affect the results. The results are discussed in terms of a conflict-monitoring account (Sturmer et al., 2002), and in terms of a feature-integration account (Hommel et al., 2002). PMID- 15114903 TI - On the replicability of the affective priming effect in the pronunciation task. AB - Bargh, Chaiken, Raymond, and Hymes (1996) and Hermans, De Houwer, and Eelen (1994) showed that a valenced target word is pronounced faster after the presentation of an affectively related prime word than after the presentation of an affectively unrelated prime word. This finding is important because it provides crucial evidence for the hypotheses that stimulus evaluation (a) is goal independent and (b) facilitates the encoding of stimuli that have the same valence. However, recent studies indicate that the affective priming effect is not a reliable finding in the standard pronunciation task. We report the results of a nearly exact replication of Bargh et al.'s (1996) Experiment 2. In line with previous replication studies, we failed to detect the affective priming effect. PMID- 15114904 TI - The size of the cross-lingual masked phonological priming effect does not depend on second language proficiency. AB - Using a masked phonological priming paradigm, Brysbaert, Van Dyck, and Van de Poel (1999) showed that Dutch-French bilinguals perform better at identifying tachistoscopically presented L2 words (e.g., oui [yes]) when those words are primed by L1 words or nonwords that are homophonic to the L2 target word according to the L1 grapheme-phoneme conversion rules (e.g., wie [who]). They noted that this priming effect was smaller for balanced bilinguals than for less proficient bilinguals, although the interaction failed to reach significance. Findings of Gollan, Forster, and Frost (1997) suggest that this could be attributed to a greater reliance on phonology in L2 reading, caused by a smaller proficiency in this language. However, in this study we show that the Dutch French cross-lingual phonological priming effect is equally large for perfectly balanced and less proficient bilinguals. Our findings are in line with more recent work of Van Wijnendaele and Brysbaert (2002). PMID- 15114905 TI - The importance of the keyword-generation method in keyword mnemonics. AB - Keyword mnemonics is under certain conditions an effective approach for learning foreign-language vocabulary. It appears to be effective for words with high image vividness but not for words with low image vividness. In this study, two experiments were performed to assess the efficacy of a new keyword-generation procedure (peer generation). In Experiment 1, a sample of 363 high-school students was randomly into four groups. The subjects were required to learn L1 equivalents of a list of 16 Latin words (8 with high image vividness, 8 with low image vividness), using a) the rote method, or the keyword method with b) keywords and images generated and supplied by the experimenter, c) keywords and images generated by themselves, or d) keywords and images previously generated by peers (i.e., subjects with similar sociodemographic characteristics). Recall was tested immediately and one week later. For high-vivideness words, recall was significantly better in the keyword groups than the rote method group. For low vividness words, learning method had no significant effect. Experiment 2 was basically identical, except that the word lists comprised 32 words (16 high vividness, 16 low-vividness). In this experiment, the peer-generated-keyword group showed significantly better recall of high-vividness words than the rote method groups and the subject generated keyword group; again, however, learning method had no significant effect on recall of low-vividness words. PMID- 15114906 TI - Levels of processing effects on implicit and explicit memory tasks: using question position to investigate the lexical-processing hypothesis. AB - One interpretation of levels of processing effects (LOP) on priming in implicit tests of memory is in terms of deficits in lexical processing during shallow study tasks. In two experiments the extent of lexical processing engaged in during standard shallow encoding tasks was manipulated by placing the encoding question either before or after the target stimulus. Clear evidence was found in explicit memory tasks that placing the question after the target stimulus increased the depth of processing of words presented in shallow encoding tasks. In contrast, there was no evidence of such an effect on the priming observed in implicit memory tasks. The results suggest that the role of lexical processing in LOP effects on priming requires further specification. PMID- 15114907 TI - Hierarchical switching with two types of judgment and two stimulus dimensions. AB - A number of recent studies have shown that shifting among four tasks that result from a factorial combination of two types of judgment with two judgment-to response mappings results in a characteristics pattern of shift costs that indicates a hierarchical representation of the two task features judgment and mapping (e.g., Kleinsorge & Heuer, 1999). In the present study, two types of judgment were combined with two stimulus dimensions. With this task combination, the same basic shift-cost pattern was observed. This suggests that a combination of two types of judgment with two stimulus dimensions also results in a hierarchically structured task representation, indicating that such a task representation is readily adopted by participants when the combination of task features is suitable for a hierarchical organization. PMID- 15114908 TI - Outcome value and early warning indications as determinants of willingness to learn from experience. AB - In this study of willingness to learn from experience, it was hypothesized that managers would show a negative outcomes bias, that is, a stronger tendency to initiate "learning-from-experience" processes after negative outcomes than after positive outcomes. Another aim of the study was to explore the impact of the existence of early warning signals about decision outcomes on the magnitude of the negative outcome bias. Eighty-three managers were asked to read vignettes describing a managerial decision and its outcomes. The outcomes were either positive or negative, and in half of the cases early warning signals existed that made it possible to predict potential negative outcomes while in the other half there were no such signals. The managers were asked to evaluate the need for a learning-from-experience process in general and to rate the degree to which several specific learning processes should be instituted in each of the scenarios. As hypothesized, a negative-outcome bias was found. The more negative the outcomes described, the stronger the managers' inclination to recommend a more intensive learning process. Similarly, a need to ensure control and follow up procedures was reported mostly after negative outcomes. The existence of early warning signals before the decision was taken did not influence the motivation to learn. Theoretical implications regarding the impact of negative outcomes in general and implications for understanding learning from experience processes in particular are discussed. PMID- 15114910 TI - What about nepotism? PMID- 15114909 TI - Help stop the killing. PMID- 15114911 TI - Does federal employment law apply to my practice? PMID- 15114912 TI - Test yourself! Are you delegating duties legally? PMID- 15114913 TI - Scholarships, and so much more. PMID- 15114914 TI - Top 10 ways to reduce no-shows and short-notice cancellations. PMID- 15114915 TI - Welcome to the new journal. PMID- 15114916 TI - AIDS threat growing throughout Europe. PMID- 15114917 TI - The IHF Journal: 75 years of serving hospitals and health services. PMID- 15114918 TI - Overview of health services in the United Arab Emirates. PMID- 15114919 TI - From research to knowledge to action: the dual challenge of health systems research. AB - In many ways, health systems research can be seen as the Cinderella of the health research world--relatively neglected by research funders, and possessing low status within the medical research community. In this short paper I wish to address two key challenges that face health systems research. The first is to achieve legitimacy as a 'proper' research area. The second is to demonstrate influence on health policy and implementation. PMID- 15114920 TI - The external assessment of health services. AB - External assessment is increasingly used worldwide to regulate, improve and market health care providers, especially hospitals. The commonest models are peer review, accreditation, statutory inspection, ISO certification and evaluation (usually internal) against the 'business excellence' framework. Each of these is progressively adapting to meet the changing demands of public accountability, clinical effectiveness and improvement of quality and safety, but the most rapid development is in accreditation. PMID- 15114921 TI - Cost control, a myth or reality: do hospital costs really go down when quality goes up? AB - Quality improvement is needed to meet the demands for quality while simultaneously reducing the cost of delivered health care. In a competitive market of corporate hospitals, quality improvement would lower the proportion of unit cost relative to potential revenue thereby increasing profit. For public hospitals, the mandate for 'minimum quality', which is the basis for accreditation, could also be attained by efficient use of hospital resources. PMID- 15114922 TI - International efforts in implementing national health information infrastructure and electronic health records. AB - Many countries are developing national strategies using information and communication technologies (ICTs) to implement health information infrastructure and electronic health records (EHR), into their medical systems. Efficiency, quality of care and medical error along with new opportunities presented by the technologies themselves have driven this process internationally. Many countries have had spectacular failures costing billions of dollars alongside some amazing successes. There has been very little dialogue internationally about what works and what doesn't work despite the fact that many government and international agencies have placed this key priority on their agendas. The nature of the technologies used promotes cooperation and these innovations in healthcare lend themselves particularly to working together for collaboration and for communication in order to learn best practice from each other. In this paper, I look at some of the national initiatives for developing an information infrastructure for healthcare as well as some of the challenges presented by these very different approaches around the world. We also review briefly the many organizations looking at international standards relating to eHealth and to implementation of electronic health records. PMID- 15114923 TI - Architecture: big dreams--small sites. AB - The explosive development of new technologies in diagnostic and treatment services and the demand for the use of these technologies by doctors and patients alike, are two strong factors that have catalysed the ongoing expansion and renovation activities among the bigger, private hospitals in the Philippines. People in respective communities and localities have demanded that their hospitals provide state-of-the-art facilities so that they need not leave their homes to use these services in far-away Metro Manila where they are provided by the more modern hospitals of the country. An example of such a hospital is a private, general and tertiary hospital in the mid-west region of the Philippines. It was founded in 1954 and has grown continuously and progressively from an eight bed clinic to the 300-bed hospital that it is now. The community around it has transformed itself from a quiet residential neighbourhood to a bustling residential-commercial area. This worked well for the hospital in terms of clientele, but the urban community has hemmed in the hospital in its site and has made physical expansion and development a difficult endeavour. The hospital administration undertook a consultative exercise among its key medical and administrative personnel and consolidated a 'wish list' based on the staff's and patients' needs and requirements perceived from day to day operations. As expected, the wish list represented the 'dream' of everyone, big dreams that are difficult to accommodate within the site of the hospital. This paper presentation will use this hospital as a model in presenting a process by which 'big dreams' may be accommodated within a 'small site'. PMID- 15114924 TI - Challenges in global blood safety. AB - Availability of safe blood and blood products is a critical component in improving health care and in preventing the transmission of infections. Millions of lives are saved each year through blood transfusion. Yet the quality and safety of blood transfusion is of continuing concern, particularly in developing countries where 82% of the world's population lives. The burden of disease and loss of life resulting from unsafe blood transfusions is devastating for communities in developing countries. WHO has developed tools and mechanisms to assess the situation of blood transfusion services globally. This provides invaluable information on the current status of the global blood supply, identifying problems and prioritizing needs and is useful in monitoring progress and trends. The issues related to the blood safety are discussed and WHO's integrated strategy for blood safety is presented in this article. PMID- 15114925 TI - Healthcare trends and challenges in Korea. PMID- 15114926 TI - The benefits of implementing a new skin care protocol in nursing homes. AB - This study explored the extent to which a new skin care protocol comprising a skin cleanser, barrier cream and barrier film could be implemented in nursing homes, and its effects on patients' skin condition, staff time and associated costs. A pre- and post-intervention study design was used. Data were collected on current skin condition and skin care procedures, before and after the introduction of a new skin care protocol. Nurses and carers were observed as they undertook skin care following episodes of incontinence. The time taken, products and amounts used were recorded. The presence and severity of incontinence dermatitis was recorded, together with the presence and severity of pressure ulceration. A supportive education programme was delivered to staff. All patients with incontinence and all staff working in six nursing homes were included in this study. Two nursing homes were randomly selected to participate in detailed skin assessments, documentation of skin care procedures and product usage. A total of 164 patients were included in detailed assessments, 79 pre-intervention, 85 post-intervention; 49 were male (29.9%) and 115 were female (70.1%). Their mean age was 83.44 years (standard deviation = 8.38). Only 3% of patients were under 70 years of age, with 72% over 80 years of age. Pre-intervention 29.1% were incontinent of urine only, 64.6% were doubly incontinent and 6.3% were catheterised. Post-intervention 29.4% had urinary incontinence, 65.9% were doubly incontinent, and 4.7% were catheterised. Skin condition was maintained or improved using the new skin care protocol. The presence of grade 1 pressure ulcers was found to significantly decrease over time (p = 0.042). The presence of incontinence dermatitis was found to be significantly lower after introducing the skin care protocol (p = 0.021). There was significant reduction in time taken to deliver skin care post-intervention (p < 0.001) with a mean time saving of 4 minutes and 2 seconds, per patient per procedure. On average this procedure was carried out 8.5 times per day, giving a time saving of 34 minutes and 17 seconds per patient per day using the new skin care protocol. This was associated with an average saving per patient per day of 8.83 Pounds for qualified staff and 3.43 Pounds for unqualified staff. Staff adherence to the new skin care protocol was good, with only one observed episode when the protocol was not followed, suggesting a high degree of success in its implementation. This study demonstrated that the introduction of a new skin care protocol, supported by an educational programme, maintained or improved patients' skin condition, and significantly reduced the resources used in delivering nursing care. PMID- 15114927 TI - Evaluation of alternating pressure air mattresses: one laboratory-based strategy. AB - Although many different type of alternating pressure air mattresses (APAMs) are used for the prevention and treatment of pressure ulcers, few high quality randomised controlled trials are available on which to base purchasing decisions. Faced with this situation, physiological measurements are increasingly being used as a surrogate. Laboratory evaluation techniques have centred largely on interface pressure (IP) measurement, typically analysing discrete maximum and minimum levels, or average pressure. However, since pressure relief is time varying, a time-based analysis technique may be more suitable for performance assessment. Measurements of IP, mattress air cell pressure (AP), skin tissue perfusion using laser Doppler fluxmetry (LDF), transcutaneous oxygen (tcPO2) and carbon dioxide (tcPCO2) were taken simultaneously on the sacrum, heels, trochanters and buttock over at least two alternating cycles. Duration of IP below three thresholds (30, 20, and 10 mmHg) as well as the area under the tcPO2, tcPCO2 and LDF curves were calculated automatically. Ten healthy volunteers were recruited to evaluate the pressure-relieving characteristics of two different designs of APAMs. Results indicated significant differences between the products. During the deflation phase of the cycle contact pressures on the heel were significantly lower (p < 0.0001) on the device whose inflation pressure was significantly higher, although there was no significant difference in deflation pressure. Therefore, it is important to note that low APs do not necessarily produce lower IPs under the heel, contrary to the intuitive classical notion. These techniques could assist in the selection of alternating or dynamic surfaces of any description confirmed by further clinical validation. PMID- 15114928 TI - Vacuum-assisted closure in the management of the open abdomen: a report of a case and initial experiences. AB - In certain surgical patients undergoing laparotomy it may be best to leave the abdomen open as a laparostomy. Care of the resulting wound represents a considerable medical and nursing challenge. This article presents such a case and the authors' initial experience of managing laparostomy wounds with vacuum assisted closure. PMID- 15114929 TI - A pilot study of the reaction forces at the heel during walking with the application of four different wound dressings. AB - A laboratory investigation was conducted to ascertain the shock-absorbing properties of four wound dressings applied to the heel area of a convenience sample of volunteers. Four subjects walked across a force plate. Data were recorded for each subject barefoot, and then while the subject had a wound care dressing stuck to the plantar aspect of their heel. Eight trials were undertaken in the five different conditions resulting in 40 trials per subject. The results suggest that one dressing, Allevyn (Smith and Nephew Healthcare Ltd, Hull, UK), when compared to other dressing materials, exhibits similar reaction forces to the normal barefoot walking data for all subjects. However, results from the anterior posterior force show no major differences across the dressings when compared to barefoot walking. Wound care dressings that have an inherent shock attenuating property may be advantageous when dealing with areas of the foot that come under excessive force. PMID- 15114930 TI - NICE guideline for pressure-relieving equipment. PMID- 15114931 TI - Using mattresses to reduce pressure on the spine. PMID- 15114932 TI - Predicting the grade of Banff 97 classification of chronic allograft nephropathy based on examination of graft dysfunction (a preliminary report). AB - OBJECTIVES: Progression of chronic allograft nephropathy (CAN) is associated with a progressive decrease in graft function. Prediction of the Banff CAN grade on the basis of correlation between the grade of histological changes and Scr is difficult because of the big spread of individual values. This study sought to predict the Banff CAN grade based on Scr, Ccr and proteinuria using ROC analysis. METHODS: Graft protocol biopsy and functional testing (Scr, Ccr and proteinuria) were performed in 77 subjects (43 men, 34 women, mean age 48.4 +/- 12.8 years) at 33.8 +/- 1.0 months after their first renal transplantation. Immunosuppression was provided with the triple combination of cyclosporin A, prednisone and azathioprine (or mycophenolate mofetil). Statistical evaluation was performed using receiver-operating curve (ROC) analysis. The cut-off value of the Banff CAN score was set at 1. RESULTS: The mean values and SD of the investigated functional parameters in study subjects were as follows: Scr = 201.5 (+/- 100.0) mumol/l Ccr = 48.1 (+/- 21.2) ml/min/1.73 m2, proteinuria = 0.89 (+/- 1.96) g/24 h. ROC analysis showed the highest AUC (+/- SEM) for Scr 0.806 (+/- 0.063). The respective values were 0.790 (+/- 0.053) for Ccr and 0.643 (+/- 0.075) for proteinuria. The AUC (area under the ROC curve) for Scr was significantly higher (P < 0.043) compared with proteinuria. The values for sensitivity (specificity) were as follows: Scr 65.0 (91.2). Ccr 75.0 (82.5), proteinuria 60.0 (68.4). The best fit values (best combination of sensitivity and specificity) were 257.2 umol/l for Scr, 33.6 ml/min/1.73 m2 for Ccr and 0.40 g/24 hr for proteinuria. CONCLUSIONS: Our findings support the assumption that Scr > 275 mumol/l and Ccr < 33.6 ml/min/1.73 m2 suggest a Banff CAN grade higher than 1 (P < 0.001). Proteinuria had the lowest predictive values. Values > 0.40 g/24 hr were probably associated with a Banff CAN grade higher than 1 (p < 0.05). PMID- 15114933 TI - Orthotopic liver transplantation for acute liver failure resulting from "acute fatty liver of pregnancy". AB - Steatosis of the liver, demonstrating itself as the acute liver failure during the third trimester of pregnancy (Acute Fatty Liver of Pregnancy--AFLP) is a rarely observed liver pathology. Herewith we describe the case of a 19-years old pregnant patient with this rare disease. The authors present the clinical course, dynamics, diagnostics, and outcome of treatment in this unique case. In this patient the ultimate treatment chosen was the liver transplantation. The opinions, concerning liver transplantation in AFLP (expressed in the world literature) are somewhat controversial. This paper presents the first case of AFLP treatment with orthotopic liver transplantation (OLTx) in Poland. PMID- 15114935 TI - Effect of ischemia/reperfusion on telomere length and CDKI genes expression in a concordant ex-vivo hemoperfusion model of primate kidneys. AB - OBJECTIVES: The telomere (T) length, p21(WAF1/CIP1) and p27(Kip1) cyclin dependent kinase inhibitor (CDKI) genes are considered the markers of cell senescence and DNA damage. The aim of the study was to evaluate the influence of renal ischemia/reperfusion (I/R) on the value of above-mentioned markers. METHODS: 13 Macaque cynomolgus monkey kidneys were harvested and placed in Eurocollins solution. 9 kidneys were ex-vivo perfused with human blood and 4 kidneys were not perfused at all (control group). Tissue expression of p21(WAF1/CIP1) and p27(Kip1) was evaluated immunohistochemically and the T lengths were measured by southern blotting technique. RESULTS: Significantly higher levels of p21 and p27 were expressed by the glomeruli (p = 0.001 and 0.0001), tubules (p = 0.0065 and 0.0006) and interstitial cells (p = 0.0017 and 0.0022, respectively) of the xenoperfused kidneys. The mean T length was higher in the control group (5.56 +/- 0.60 kbp) than in the study group kidneys (5.46 +/ 0.36 kbp) (P = NS). CONCLUSION: Renal I/R is associated with telomere shortening and an over-expression of p21 and p27 genes indicating substantial DNA damage and/or accelerated tissue senescence. PMID- 15114934 TI - Donor-specific antibodies and kidney transplant rejection. AB - The role played by non-complement binding antibodies after kidney transplantation is still not satisfactorily elucidated. A growing number of publications suggest that the detection of antibodies to donor HLA antigens by more sensitive methods than the standard complement-dependent cytotoxicity test is often associated with acute and chronic immunological complications and generally worse graft prognosis. In this review, we briefly discuss several recent reports on the clinical significance of non-cytotoxic anti-HLA antibodies, which might have important implications for patient immunosuppression and management after organ transplantation. PMID- 15114936 TI - No evidence for productive PERV infection of baboon cells in in vivo infection model. AB - OBJECTIVES: The discovery that pig endogenous retroviruses are infectious for human cells in vitro lead to vehement discussions about the possible risk of infection after clinical xenotransplantation. Since PERV transmission to non human primate cells in vitro has been observed, similar to human cells, infection studies in non-human primates should represent the best model to analyze a potential PERV transmission after xenotransplantation. However, it is still open to discussion, whether non-human primate cells can be infected productively similar to human cells- and whether those species are suitable to analyze PERV infection risks in vivo. METHODS: In vitro, only few cell types can be tested for susceptibility. We developed a pig to baboon cell transplantation model with special emphasis on B-cell effective immunosuppression, removal of anti Gal-alpha 1,3-Gal-antibodies, inhibition of the complement cascade and long term survival of transplanted cellular grafts. This model allows us to investigate in vivo, whether any baboon cell types may be permissive for productive PERV infection. The xenograft recipients were investigated for up to 535 days post transplantation. Gal-alpha 1,3-Gal-antibody and complement levels were monitored. Potential PERV transmission was analyzed, not only in PBMC, but in a variety of tissue samples as well as in serum and plasma samples by PCR, RT-PCR and by detection of RT-activity. Moreover, potential PERV specific immune responses were studied by a highly sensitive Western-Blot-assay. RESULTS: Despite several days of extremely low levels of Gal-alpha 1,3-Gal-antibody and complement, and despite of long term xenochimerism, no evidence for PERV infection was obtained in any of the tested tissues or in the tested serum samples. CONCLUSION: This study supplies further evidence for a low susceptibility of baboons towards productive PERV infection after xenotransplantation. PMID- 15114937 TI - Modulation of human anti-pig T cell responses by monoclonal antibodies directed to porcine CD45 molecules. AB - Antibody-mediated targeting of pig costimulatory molecules is assumed to be a possible strategy to achieve donor-specific tolerance after xenotransplantation. However, porcine molecules of the B7 family (e.g. CD86) are expressed on typical antigen presenting cells (APC) and also on vascular endothelial cells. Thus, in vascularized porcine xenografts the usage of therapeutic anti-B7 monoclonal antibodies (mAb) might be associated with damage of the endothelium. OBJECTIVE: In the present study we asked whether modulation of human T cell reactivity can be obtained by targeting molecules selectively expressed on pig leucocytes. METHODS: MAb directed to pig CD45 were tested for their capacity to modulate the in vitro activation of human T cells induced by porcine peripheral blood mononuclear cells. RESULTS: Porcine stimulatory cells induced significant proliferation of human T cells. In the presence of porcine CD45 mAb human T cell responses were reduced by 30-40%. The inhibitory effects were most pronounced when CD45RA mAb were used whereas mAb directed to CD45RC isoforms only moderately inhibited human T cell activation. The tested antibodies had no effects on human T cell activation induced by mitogens or by alloantigen. CONCLUSION: Manipulation of CD45 molecules on pig leucocytes may reduce their potential to stimulate human T cells. In recipients of vascularized porcine xenografts the usage of anti-pig CD45 mAb could be an approach to block the direct pathway of T cell activation initiated by porcine APC without affecting the endothelium of the graft. PMID- 15114938 TI - Porcine endogenous retroviruses (PERVs) and xenotransplantation: screening for transmission in several clinical trials and in experimental models using non human primates. AB - Xenotransplantation may develop into a medical technology able to save or improve the quality of life. Porcine endogenous retroviruses (PERVs), because they are integrated in the genome of all pig strains, because they are produced by normal pig cells, and because they can infect human cell in vitro, are considered to be the main microbiological risk if pig cells, tissues or organs are to be transplanted. Indeed, serial passaging of PERV on human cells, simulating the situation during xenotransplantation, was found to increase the titer of the virus and was associated with corresponding genetic changes in the viral LTR. In vitro infection studies showed a productive infection of primary cells of different species including non-human primates and man. However, using newly developed sensitive detection methods, evidence for PERV transmission was seen neither in over 200 patients who had received porcine xenotransplants nor in butchers frequently exposed to pig tissues. Similarly, rats, guinea pigs, minks, rhesus macaques, pig tailed macaques and baboons inoculated with high doses of PERV and given strong daily immunosuppressive treatment failed to exhibit evidence of infection. These data are crucial for the evaluation of xenotransplantation safety because they demonstrate that PERVs cannot easily be transmitted to other species including man. PMID- 15114939 TI - Morphological assessment of reperfusion injury in a concordant xenogeneic primate model. AB - Ischemia and consecutive reperfusion injury of the graft cannot be avoided in organ transplantation. One principal characteristic of this process is the temporary and permanent adherence of leukocytes to endothelial cells of the graft as well as damage of the associated tissue. Polyclonal antithymocyte globulins (ATGs) are used to prevent acute rejection after transplantation and to overcome graft vs. host disease. ATGs induce apoptosis and complement-mediated cell death in peripheral T-lymphocytes having the potential to inhibit leukocyte adhesion by direct binding to adhesion molecules. The aim of the present study was to analyse in a non-human primate model (cynomolgus monkeys) morphological changes within the microvasculature and the different cell-subpopulations upon ischemia/reperfusion after immunosuppressive treatment. Our results show a decrease of the tissue damage and WBC infiltration in muscular structures as well as a lower number of mononuclear cells in peripheral blood in the ATG-treated group compared to a non-treated control group. We may conclude that pATGs have a beneficial effect on the early mechanisms of ischemia/reperfusion injury. PMID- 15114940 TI - The fate of the Council of Europe's recommendation on xenotransplantation. AB - The draft of the working group of experts for a Recommendation of the Committee of Ministers to Member States on Xenotransplantation was finalized in September 2001, ending a working period of two and a half years. This paper will briefly reflect upon the development of the recommendation up to this point in time. A selection of specific articles in the document, which played a significant role in its creation, will be explained in greater detail. The last section addresses the most recent developments with regard to the Draft Recommendation and sketches out the way towards its finalization. PMID- 15114941 TI - Retroviral safety: analyses of phylogeny, prevalence and polymorphisms of porcine endogenous retroviruses. AB - Porcine endogenous retroviruses (PERV) are discussed as putative infectious agents in xenotransplantation (XTx). PERV classes A, B, and C harboring different envelope proteins and two types of long terminal repeat (LTR) structures exist. One type of LTR contains a distinct repeat structure in U3, while the other is repeat-less and confers poor transcriptional activity. As the different LTR structures were found to be distributed unequally among the proviruses, we were interested in determining which LTR is the ancestor. Since replication-competent PERV can still be found today suggesting an evolutionary recent origin, we investigated the distribution and prevalence of six well-characterized and chromosomally assigned PERV in individuals of five different pig (Suidae, Sus scrofa) subspecies. Our studies revealed a heterogenous distribution of replication-competent PERV among individuals as well as among subspecies. The age of PERV was calculated to be 7.6 x 10(6) years, whereby the repeat-less LTR type evolved approximately 3.4 x 10(6) years ago being the phylogenetically younger structure. The age correlates with the time of separation between pigs and their closest relatives, american-borne peccaries (Tayassuidae, Pecari tajacu), 7.4 x 10(6) years ago. PMID- 15114942 TI - Common factors and our sacred models. AB - In this article we argue that much of what makes one treatment effective is common to other forms of effective treatment--both in psychotherapy generally and in marital and family therapy (MFT) specifically. Yet MFT has largely ignored the research on common factors. In this article we present a moderate view of common factors that, while repudiating the extreme position that there is no difference among treatment models, stresses that there are common factors and mechanisms of change that undergird most forms of successful treatment. These common mechanisms of change should be given more attention in our field, which has tended to emphasize the uniqueness of our sacred models. We delineate some of the major common factors, review the empirical evidence for them, and discuss implications of adapting a common factors informed approach to family therapy. PMID- 15114943 TI - Beyond common factors: multilevel-process models of therapeutic change in marriage and family therapy. AB - A number of scholars have proposed the common factors perspective as the future direction of marriage and family therapy (MFT). Although intuitively appealing, the case for the common factors perspective is not as clear-cut as proponents portray. In its current form, the common factors perspective overlooks the multilevel nature of practice, the diversity of clients and settings, and the complexity of therapeutic change. In contrast, comprehensive process-based change models are an alternative to the limitations of common factors. In this article, we consider the limitations of the common factors perspective and propose the necessary and sufficient components and processes that might comprise comprehensive, multilevel, process-based therapeutic change models in MFT. PMID- 15114944 TI - Common factors are not islands--they work through models: a response to Sexton, Ridley, and Kleiner. AB - In this article, we respond to Sexton, Ridley, and Kleiner (this issue) from three different perspectives. First, we discuss their criticisms as rooted in a portrait of common factors to which we do not subscribe. Second, we discuss points of agreement and partial agreement between our two articles. Finally, we discuss our areas of clear disagreement with their points of view. In these areas of disagreement we put forth the common factors approach as an empirically supported lens; we discuss the influence of investigator allegiance on the specificity conclusion; and we challenge the idea that the common factors lens is simple. In conclusion, we illustrate how common factors work through a credible therapy model using functional family therapy as an example. PMID- 15114945 TI - Implications of a moderated common factors approach: does it move the field forward? AB - There are a number of similarities between the moderated common factors proposal of Sprenkle and Blow (this issue) and the multilevel, process-based therapeutic change presentation of Sexton, Ridley, and Kleiner (this issue). Despite these areas of agreement there are fundamental differences in our respective positions. We suspect that these differences are not unique to the respective authors, but instead represent the current debate in the field regarding common factors. In this commentary, we discuss what we think are important issues to have emerged from these articles. Our hope is that the issues we present below will take the debate and discussion one step further. PMID- 15114946 TI - Getting reluctant substance abusers to engage in treatment/self-help: a review of outcomes and clinical options. AB - This review examines the results from 19 outcome studies, involving 1,501 cases, within 10 different clinical approaches to getting alcohol or drug abusers to engage in treatment or self-help. Each approach is summarized and its "success rate(s)" presented. Comparisons are made across various subcategories, such as alcohol vs. drug abusers, adolescents vs. adults, and experimental conditions vs. controls. A distinction is drawn between "Dual-Purpose" approaches, which combine engagement with counseling of the concerned person who originally sought help for the substance abuser, and "Engagement-Primary" approaches, which generally confine their efforts (through working with family members and others) to getting the substance abuser into treatment/self-help. "Best bet" clinical options are presented in terms of which particular approach appears to have the edge with which kind of substance abuser. PMID- 15114947 TI - Evidence-based practice for marriage and family therapists. AB - The purpose of this article is to integrate science into clinical practice by introducing marriage and family therapists to the ideas of evidence-based practice (EBP). Evidence-based practice, which originated in the medical field, refers to the process of using research to make clinical decisions that best meet the needs of each client. Included in the description is a brief history of EBPs and ideas about learning EBPs. Suggestions are also made about the use of EBPs in MFT training programs, and resources are provided to enable clinicians to use EBPs in clinical practice. PMID- 15114948 TI - Affection and conflict in marital and parent-child relationships. AB - In this study, we examined affection and conflict in marital and parent-child relationships, as reported by mothers, fathers, and 9- and 10-year-old children in a community sample of 87 families. Affection and conflict were inversely related within relationships, with mixed findings across relationships. Most reports showed an association between marital and parent-child conflict as well as between marital and parent-child affection. According to fathers, however, the association between marital affection and father-child affection is moderated by marital conflict. Clinical implications of the associations between positive and negative dimensions of family interaction and of the links between marital and parent-child relations are discussed. PMID- 15114949 TI - An integrative intervention for promoting recovery from extramarital affairs. AB - The discovery or disclosure of an extramarital affair can have a devastating impact on partners, both individually and on the relationships. Research suggests that affairs occur relatively frequently in relationships and are a common presenting problem in couple therapy. However, despite their prevalence, there is little empirical treatment research in this area, and most therapists describe this problem as one of the more difficult to treat. In this study, we used a replicated case-study design to explore the efficacy of an integrative treatment designed to help couples recover from an affair. Six couples entered and completed treatment. The majority of these couples were less emotionally or maritally distressed at the end of treatment, and the injured partners reported greater forgiveness regarding the affair. Details of the intervention, suggested adaptations of the treatment, and areas for future research are discussed. PMID- 15114950 TI - Toward a mini-theory of the blamer softening event: tracking the moment-by-moment process. AB - In this article we present the results of a discovery-oriented task analysis research study identifying specific therapist behaviors that facilitate softening events in emotionally focused therapy (EFT) for couples. Therapy sessions conducted by an expert EFT therapist were examined. Six therapist content theme shifts were identified and specific interventions were delineated within each using an EFT coding scheme (EFT-CS) created for this study. This study extends the theoretical understanding of the softening process--a key change event in an empirically validated couples therapy approach--and provides a detailed clinical map for the training of therapists. PMID- 15114951 TI - Measles vaccines. PMID- 15114952 TI - Influenza in the world. PMID- 15114953 TI - Avian influenza A(H5N1)--situation (poultry) in Asia as at 2 March 2004: need for a long-term response, comparison with previous outbreaks. PMID- 15114954 TI - Ninth informal consultation on the global polio laboratory network, 8-10 September 2003, WHO headquarters, Geneva, Switzerland. PMID- 15114955 TI - WHO consultation on priority public health interventions before and during an influenza pandemic, Geneva, 16-18 March 2004. PMID- 15114956 TI - Cardiac technology goes to the wall. A new support device may help when other treatment options run out. PMID- 15114957 TI - When "catching a chill" might save your life. PMID- 15114958 TI - "Stealth" fat will be revealed on labels in 2006. PMID- 15114959 TI - Cholesterol drugs may help prevent errant heart rhythms. PMID- 15114960 TI - Stent cases raise a caution flag. Patients can help ensure safe use of new drug coated devices. PMID- 15114961 TI - The ABCs of EKGs and other tests. These key diagnostic tools often help set the course for treatments. PMID- 15114962 TI - The path of healthy resistance. Why some patients pull, push--and break an old taboo. PMID- 15114963 TI - I have been diagnosed with heart failure, and my cardiologist has prescribed a medication called Coreg. What will it do for me? PMID- 15114964 TI - Is it possible to replace heart valves without surgery? I think I've heard that this was being done in Europe for patients with aortic stenosis. PMID- 15114965 TI - A friend of mine is being considered for a heart transplant. How successful are heart transplants today? PMID- 15114966 TI - When your heart sends an early SOS. How to get just-in-time help for 3 under diagnosed cardiac conditions. PMID- 15114967 TI - New light on a night-time breathing disorder. PMID- 15114968 TI - Where there's smoke, there's inflammation. PMID- 15114969 TI - The case (continued) against vitamin supplements. PMID- 15114970 TI - A cholesterol drug protects some people with diabetes. PMID- 15114971 TI - 2 drugs team up for protection. Old standby joins relative newcomer to battle heart disease. PMID- 15114972 TI - Relief for the overworked heart. A "thick heart" can be a sick heart, but treatments can help. PMID- 15114973 TI - "Space suit" boosts circulation. For once, high pressure helps--to improve quality of life. PMID- 15114974 TI - I am a 70-year-old man, and my internist recommended an ultrasound to look for an aneurysm in my belly. Do you think this is necessary? PMID- 15114975 TI - I saw on television that patients with heart failure may be candidates for a form of therapy called cardiac resynchronization. What is this procedure, and is it safe? PMID- 15114976 TI - Is there a link between erectile dysfunction and heart disease? I read this in a men's magazine, but I don't see how the two conditions can be connected. PMID- 15114977 TI - Heart care counters multiple threats. Preventive cardiology also cuts risk of arthritis, cancer and diabetes. PMID- 15114978 TI - "Super aspirin" cuts risks after multiple cardiac events. PMID- 15114979 TI - Homing in on ways to build new blood vessels. PMID- 15114980 TI - Drinking wine, beer or liquor may douse inflammation. PMID- 15114981 TI - New drug fights fluid build-up. Medication may relieve a common heart failure symptom. PMID- 15114982 TI - 4 new tests for heart disease. Here's what your doctor might order in the future. PMID- 15114983 TI - Can you catch cardiac disease? Here's what to do while doctors sort out conflicting answers. PMID- 15114984 TI - Which type of open heart surgery is better, on the bypass machine or off? I want to know which one to ask for. PMID- 15114985 TI - After a heart attack, my doctor started me on cholesterol-lowering medications. My last cholesterol check showed it to be very low. Is this safe? PMID- 15114986 TI - I am sick of hearing about the obesity epidemic in this country. Aren't the news media just exaggerating? Is there really a problem? PMID- 15114988 TI - Debate over ritonavir price increase gains momentum. Critics seek to reverse the fivefold price hike through legal action and boycott of Abbott's products. PMID- 15114987 TI - Dislocation of the mandible. AB - We report the first case of bilateral temporomandibular joint (TMJ) dislocation in a woman after endoscopy. We go on to discuss the case reports published in the literature, with particular focus on potential factors that may predispose to this complication, and we offer some suggestions on dealing with this significant complication. PMID- 15114989 TI - Court rules girl can take sex-change therapy. PMID- 15114990 TI - Wildfire coverage. PMID- 15114991 TI - Why no hand-grips? PMID- 15114992 TI - Shanghai to allow divorcees a second child. PMID- 15114993 TI - A 'GEM' of a program. PMID- 15114994 TI - Letter leaves bitter taste. PMID- 15114995 TI - Doctors in Peru continue strike despite budget increase. PMID- 15114996 TI - Uganda considers DDT to protect homes from malaria. Health officials claim DDT will help save money, but critics warn of environmental costs. PMID- 15114997 TI - Career opportunities in organ procurement. PMID- 15114998 TI - EEOC v. BNSF: the risks and rewards of genetic exceptionalism. PMID- 15114999 TI - Curbing misuse of genetic information in light of the Enron debacle: a comment on Dean Partlett's proposal. PMID- 15115000 TI - Pushing drugs: genomics and genetics, the pharmaceutical industry, and the law of negligence. PMID- 15115001 TI - Penetration of the small intestine of a California sea lion (Zalophus californianus) pup by adult hookworms (Uncinaria spp). AB - During a study on the mortality of California sea lion (Zalophus californianus) pups born on San Miguel Island, California in 2002, two adult female hookworms (Uncinaria spp) were found penetrating the serosal surface of the intestinal wall and protruding into the peritoneal cavity of one pup. Documentation and a description of this unexpected finding and associated lesions are presented here. Also, adult hookworms were found in the peritoneal fluid of two other dead Z. californianus pups. PMID- 15115002 TI - Causes for concern and causes of action: a comment on "Pushing drugs". PMID- 15115003 TI - Genomic torts: a response to Professor Feldman. PMID- 15115004 TI - Treatment of gallstone and gallbladder disease. SSAT patient care guidelines. PMID- 15115005 TI - Surgical repair of groin hernias. SSAT patient care guidelines. PMID- 15115006 TI - Esophageal achalasia. SSAT patient care guidelines. PMID- 15115007 TI - Surgical repair of incisional hernias. SSAT patient care guidelines. PMID- 15115008 TI - Endoscopic fibrin sealing of congenital pyriform sinus fistula. AB - Pyriform sinus fistula is a very rare branchial apparatus malformation, often appearing in the form of a cervical inflammatory process (abscess or suppurative thyroiditis), especially in infants. Failure to diagnose this lesion may result in unexpected recurrence. A case of recurrent suppurative thyroiditis caused by pyriform sinus fistula in a 9-year-old girl is reported. In the latency period of infection, the fistula tract was identified by a barium meal contrast study. Direct endoscopy showed the fistula internal orifice at the apex of the left pyriform fossa. The fistula was completely obliterated by injection of fibrin glue. Suppurative thyroiditis is reported mainly in the pediatric literature, and the reported case is the first to be managed endoscopically by injection of fibrin adhesive. PMID- 15115009 TI - A sketch of L'avenir de la nature humaine. PMID- 15115010 TI - Remarks on Habermas's presentation of "L'avenir de la nature humaine". PMID- 15115011 TI - Physicians' conflicts of interest. PMID- 15115012 TI - Jejunal leiomyosarcoma, a rare cause of obscure gastrointestinal bleeding diagnosed by wireless capsule endoscopy. AB - Obscure gastrointestinal bleeding is a relatively frequent disorder and may account for as many as 5% of all cases of gastrointestinal bleeding. The etiology of these hemorrhages may be attributed to lesions in the small intestine, which may not show up in radiologic studies, located in areas inaccessible to conventional endoscopy. The case of a 50-year-old patient admitted to the hospital on two occasions for gastrointestinal bleeding requiring blood transfusions is reported. On the first occasion, the bleeding was thought to be caused by a duodenal ulcer because no other lesions prone to bleeding were found. At the next admission for recurrent bleeding, the ulcer was found to have healed and thus was ruled out as the cause. Wireless capsule endoscopy detected an ulcerated tumor invading the submucosa of the jejunum. The pathologic diagnosis was low-grade leiomyosarcoma. Wireless capsule endoscopy has proved to be far superior to other radiologic and endoscopic techniques for the diagnosis of obscure gastrointestinal bleeding and pathologies of the small intestine in general. PMID- 15115013 TI - Ex post facto IRB review: two practical hurdles, one conceptual mistake. PMID- 15115014 TI - Laparoscopic repair of an internal strangulated supravesical hernia. PMID- 15115015 TI - Laparoscopic resection of a pancreatic polypeptidoma with a solitary liver metastasis. AB - Pancreatic neuroendocrine tumor is an uncommon disease, and surgery is the only potentially curative treatment even when there is hepatic metastasis. A patient undergoing concomitant laparoscopic distal pancreatectomy and hepatectomy for pancreatic polypeptidoma with a solitary liver metastasis is reported. PMID- 15115016 TI - Laparoscopic splenectomy for the treatment of wandering spleen. AB - Wandering spleen is a very rare pathologic condition that initially presents with unremarkable clinical symptoms, then dramatically manifests severe complications. A case of wandering spleen treated with laparoscopic splenectomy is described and compared with other similar reports in the literature. Laparoscopic exploration of the abdominal cavity allowed the diagnosis to be confirmed and splenectomy to be performed. An enlarged spleen led to the choice of total splenectomy to avoid both future complications and future organ torsion resulting from the long and twisted vascular pedicle. The laparoscopic approach led to a rapid recovery, a 3 day postoperative course, and satisfactory cosmetic results. PMID- 15115017 TI - Minimizing harm in agricultural animal experiments in New Zealand. AB - Intrusive agricultural experiments published in New Zealand in the last five years are reviewed in terms of the degree of animal suffering involved, and the necessity for this suffering in relation to research findings. When measured against animal welfare criteria of the Ministry of Agriculture, thirty-six studies inflicted "severe" or "very severe" suffering. Many of these experiments had questionable short-term applications, had an application restricted to agricultural production or economic growth, or could have been modified to prevent or reduce suffering. PMID- 15115018 TI - Reasoning about embryos, cloning and stem cells: let's get more clear and distinct. AB - Plural democratic societies encourage and require the tolerance of disparate views. However, in relation to contentious areas like assisted reproductive technologies and destructive embryo research, tolerance is strained by the normative force of our fundamental beliefs about the moral status of early human forms. Yet in the continuing debates, spokespersons for different positions often do not concede all the implications of their arguments, may sidestep the real moral issues, and can fail to be clear about the foundations on which their arguments and policy advice ultimately rely. Guidelines and statutes can be rendered incoherent by the desire to balance and satisfy opposing values, rather than honestly reflecting the primary values they espouse. I call for greater clarity and honesty as these issues continue to be debated. An uncritical adherence to pluralism will encourage strategic obfuscation, but citizens of democracies need to be clearly informed about all the premises of opposing positions. Decisions about ethically and legally acceptable reproductive technologies ultimately depend on support for one metaphysical grounding at the expense of another. This should be acknowledged, as should its implications for policy. PMID- 15115019 TI - Response to Malcolm Parker. PMID- 15115020 TI - Commentary on Malcolm Parker. PMID- 15115021 TI - Insurance companies' access to genetic information: why regulation alone is not enough. AB - The background of this paper is the ongoing dismantling of the social insurance systems in favour of commercialisation and privatisation of insurances needed for illness, old age and premature death. This combined with the increased possibility of using genetic testing for differentiating personal insurance premiums has the potentiality of creating a 'genetic proletariat'--an uninsurable high-risk population. The common way of handling this problem in Sweden, and many other developed countries around the North Atlantic, has been to regulate insurance companies' right to ask for and use genetic information in various ways. There is a distinction between partial regulation (that allows insurance companies access to genetic information from genetic tests already made, sometimes only above a specified amount, but not to demand new tests) and total regulation (that forbids insurance companies to ask for or use any genetic information). I will argue that these forms of regulation probably will have adverse consequences given the dismantling of collective social insurance systems. If this is convincing, a better way to solve the problem of an uninsurable high-risk population (and other problems) is to resurrect the collective, obligatory insurance systems in which the individual risk profile does not constitute a basis for premium determination. Both arguments cast in terms of consequences and justice render support for this conclusion. PMID- 15115022 TI - Secret ethics business? AB - In this paper, I question the common assumption that the workings of Human Research Ethics Committees should be treated as confidential. This is actually quite a complex issue, since there are many stages in the ethics approval process, and a number of different stakeholders who might wish to claim access, or restrict access, to different sorts of information. Here I consider just one aspect--whether ethics committee members should be free to reveal in public the details of what occurs in their meetings. My approach is two-fold: first a negative argument that confidentiality does not apply to ethics committee deliberations, and then a positive argument that there is an important public good, namely accountability, to be served by making these deliberations open to the public. PMID- 15115023 TI - Benefits of participation in longitudinal qualitative research study. AB - Although mainstream research institutions and health care organisations are now starting to acknowledge the important contribution of qualitative research, there are still many obstacles to obtaining funding. Consequently, at all points along the continuum of obtaining funds, enrolling participants and conducting the research, qualitative researchers will benefit from being able to refer to, or reference, a body of empirical knowledge that addresses ethical issues raised by those who have responsibility for decision-making about the implementation of research proposals. This article has been written with this strategy in mind. One such ethical issue is a concern about the impact on participants of being involved in qualitative studies when they are in a vulnerable state from a distressing life experience. This article provides empirical data that describes the benefits associated with qualitative research process for parents enrolled in a longitudinal study looking at the issues associated with diagnosis and treatment for paediatric haematological malignancies. The empirical findings affirm the significant beneficial contribution that qualitative research can make to the lives of those coping with an extremely challenging health care situation. PMID- 15115024 TI - 'Science is really needed--that's all I know': informed consent and the non verbal practices of collecting blood for genetic research in northern Sweden. AB - In Vasterbotten County in northern Sweden a start-up biotech company has recently gained all commercial rights to one of the worlds largest population based research biobanks. The biobank and the company have publicly emphasized that all donors have given their informed consent to participate, but within the academy it has become debated whether people have been adequately informed. Based on anthropological fieldwork it is shown that many people do not read the information provided. The data do not, however, suggest that donors themselves perceive a lack of information. This article endeavours to make meaningful the apparent lack of interest among donors in the information they are offered. It is argued that the donation of blood should be analysed in its social and historical context rather than as a response to rational assessment of information of research purposes. It implies a conceptualisation of agency more aware of the intersubjective nature of moral negotiation than usually implied in studies of informed consent. PMID- 15115025 TI - Genetic testing and primary care: a new ethic for a new setting. AB - For several decades, clinical geneticists have espoused two key ethical principles, nondirectiveness and confidentiality. These principles made a great deal of sense in the highly personal and controversial setting of reproductive genetics. Now that clinical genetics has entered the primary care setting, clinicians are rethinking the strength of their commitment to these traditional norms and they are revamping their ethical priorities. Patients increasingly need advice about whether they should take genetic tests and whether and how they should respond to the test results. Patients also need to know about how this information will impact family members and whether other members of their family should be tested. Clinical geneticists may even consider breaking individual confidentiality in order to prevent harms to family members. Although clinical geneticists do not need to abandon nondirectiveness and confidentiality in this new setting, they may not strictly adhere to these principles in some circumstances in order to benefit patients and their families. PMID- 15115026 TI - Learning from the voiceless. AB - This article is concerned with understanding what is at stake in the everyday lives of family members facing Huntington's Disease (HD). The methodological and analytical point of departure is German critical psychology, particularly the category of conduct of everyday life (Holzkamp, 1995; Dreier, 1999). Specifically, I address questions of accessing and understanding the conduct of everyday life of persons facing HD who are not visibly active with respect to this circumstance. The question of access is not merely about getting in touch with persons who are not known to the research, professional and HD communities, but also about the consequences of establishing contact with persons who have not made an entry into any of these public areanas themselves. The question of understanding is about developing an analysis from a first-person perspective on the personal conduct of everyday life that is not visibly active. The development of such an understanding has broader implications, not just for further research and health care practices, but importantly also for the prevailing moral and ethical demands made on persons living at risk of hereditary diseases. PMID- 15115027 TI - Right hepatic artery pseudoaneurysm and cystic duct leak after laparoscopic cholecystectomy. AB - Laparoscopic cholecystectomy (LC) seems to be associated with an increased risk of biliary or vascular injuries. Hepatic artery pseudoaneurysms (HAP) are rare complications of LC. HAP can occur in the early or late postoperative period. Patients with HAP present with abdominal pain, hemobilia, and liver function test (LFT) alterations. We report the case of a patient who was affected with a cystic duct stump leak associated with a right HAP and was treated by endoscopic biliary drainage and angiographic coil embolization. PMID- 15115028 TI - Transgastric laparoscopic approach for resection of hemorrhagic Dieulafoy's vascular malformation. AB - We report the case of a successfully resected hemorrhagic Dieulafoy's vascular malformation using a transgastric laparoscopic approach. A 37-year-old man was admitted for hematemesis and anemia. Emergency endoscopy revealed active hemorrhage from a submucosal vascular malformation in the posterior gastric wall. Dieulafoy's disease was suspected. It was initially treated by repeated endoscopy. Due to recurrent bleeding, on day 3 the patient was scheduled for surgery. The laparoscopic treatment consisted of an anterior gastrotomy, followed by extirpation and resection of the lesion after evaluation of the gastro esophageal junction. There was no need for intraoperative endoscopy. To our knowledge, this is the first description of the application of a transgastric laparoscopic approach for the management of a hemorrhagic Dieulafoy's vascular malformation. PMID- 15115029 TI - Clinical predictors of leak after laparoscopic Roux-en Y gastric bypass for morbid obesity. PMID- 15115030 TI - Facilitation of open Spigelian hernia repair by laparoscopic location of the hernial defect. PMID- 15115031 TI - Burning the candle at both ends, and there is nothing left for proof: the Americans with Disabilities Act's disservice to persons with mental illness. PMID- 15115032 TI - Much ado about gene patents: the role of foreseeability. PMID- 15115033 TI - Sonographic evaluation for peripheral pulmonary nodules during video-assisted thoracoscopic surgery. PMID- 15115034 TI - Actor-network theory: a tool to support ethical analysis of commercial genetic testing. AB - Social, ethical and policy analysis of the issues arising from gene patenting and commercial genetic testing is enhanced by the application of science and technology studies, and Actor-Network Theory (ANT) in particular. We suggest the potential for transferring ANT's flexible nature to an applied heuristic methodology for gathering empirical information and for analysing the complex networks involved in the development of genetic technologies. Three concepts are explored in this paper--actor-networks, translation, and drift--and applied to the case of Myriad Genetics and their commercial BRACAnalysis genetic susceptibility test for hereditary breast cancer. Treating this test as an active participant in socio-technical networks clarifies the extent to which it interacts with, shapes and is shaped by people, other technologies, and institutions. Such an understanding enables more sophisticated and nuanced technology assessment, academic analysis, as well as public debate about the social, ethical and policy implications of the commercialization of new genetic technologies. PMID- 15115035 TI - Biological weapons, genetics, and social analysis: emerging responses, emerging issues--II. AB - Recent terrorist attacks in the United States have generated significant attention in many countries to the threats posed by biological weapons. In response to these events and the specter of future attacks, bioscientists and professional organizations have begun or intensified asking questions about the possible malign applications of their research. Part II of this two-part article examines the emerging responses initiated by biomedical organizations and spokespersons in the US and the UK. In doing so it considers how scientific and medical research communities are defining and policing notions of professionalism, responsibility and accountability in the responses made. Through an examination of these issues, suggested lines for future social analysis are offered. PMID- 15115036 TI - Codes of ethics: some history. PMID- 15115037 TI - Advertising policies of medical journals. PMID- 15115038 TI - Ethics in phase II trials of cancer therapies. PMID- 15115039 TI - Clinical trials of cancer therapy: a personal narrative. PMID- 15115040 TI - Clinical trials of cancer therapy: a social worker's view. PMID- 15115041 TI - The importance of knowing where you are. PMID- 15115042 TI - Cancer research and therapy: a view from Norway. PMID- 15115043 TI - In the face of death: four ethical issues. PMID- 15115044 TI - Monash University Centre for Human Bioethics. PMID- 15115045 TI - A global ethics institute in New Zealand and Japan. PMID- 15115049 TI - Emergency medicine. PMID- 15115050 TI - Deep brain stimulation. PMID- 15115051 TI - Earlier onset of treatment or increment in LT4 dose in screened congenital hypothyroidism: which as the more important factor for IQ at 7 years? AB - OBJECTIVE: To determine between timing and LT4 dose which was the more important factor for IQ at 7 years in screened congenital hypothyroidism (CH). METHODS: 131 children with CH born from 1979 to 1994 and 30 controls were studied. Mean age at recall: 22.8+/-1.1 days. Mean initial LT4:5.6+/-0.1 microg/kg/day. RESULTS: Optimal global IQ (GIQ; 119+/-1.8) was obtained for a recall < or =15 days. Results for a recall after 3 weeks were lower (107.7 +/- 2.4). The IQ of infants treated before 21 days (117.1 +/-1.2) was identical to the IQ of those treated after this threshold was lower (108.6 +/- 1.7). No significant differences for GIQ were observed with various initial LT4. Infants treated with a dose of LT4 > or = 6 microg/kg/day had a higher performance IQ (117.3 +/-1.8 vs. 112.8+/- 1.2) compared with those treated with a dose < 6 microg/kg/day. The severity of CH and socio-economic levels were similar in all groups. CONCLUSION: In this study, timing appears to be more important factor for the intellectual outcome. PMID- 15115053 TI - Current awareness in prenatal diagnosis. PMID- 15115052 TI - Risky business: when mom and pop buy health insurance for their employees. AB - The economics of small group insurance makes offering health benefits to employees a risky business. Surveys of employers from 1989 to 2003 reveal that more rapid premium increases are forcing small firms to impose higher cost sharing. In 2003, premiums for small firms (3-199 workers) increased 15.5 percent, outpacing the 13.2 percent increase for large firms (200+ workers). From 2000 to 2003, deductibles among small firms increased 100 percent in PPO plans when employees use in-network providers and 131 percent when they use out-of network providers; among large firms, deductibles in PPO plans increased 33 percent and 44 percent, respectively. And in 2003, 40.3 percent of employees in the smallest firms contributed 41 percent or more of the total family premium, compared with only 11.2 percent of employees in large firms. Clearly, fundamental change in the small employer market is necessary, including new options for helping small firms gain access to the advantages large firms have in purchasing health benefits. PMID- 15115055 TI - Research on deafness. Doctoral dissertations. PMID- 15115054 TI - A national research agenda for the postsecondary education of deaf and hard of hearing students: a road map for the future. AB - This article describes converging nationwide changes in the postsecondary education of students with hearing loss during the past 30 years. Simultaneous trends in the economy, labor force, and business practices have magnified the need for literacy, postsecondary training, and career skills. These conditions stimulated institutional and professional activities that led to drafting a National Research Agenda report to guide development of federally funded research projects in postsecondary education. These studies will enhance better understanding of the complex interactions of diverse support services, learning living environments, and student populations in a broad continuum of post-high school vocational and academic training programs. The conceptual framework of the Agenda is explained, as are its expected goals, criteria for research projects, benefits, and outcomes. This article interweaves the perspectives and roles of postsecondary and vocational rehabilitation professionals, federal officials, and researchers contributing to the preparation of the Agenda report. Relevant national research studies are cited and consumer involvement in research is emphasized. PMID- 15115056 TI - Clinical quiz. Calcified synovial sarcoma. PMID- 15115057 TI - Cam Perry, Heraclitus, and hypnosis: an appreciative understanding. AB - The authors summarize personal, intellectual, and social influences on Campbell Perry's (1937-2003) life and research on hypnosis. His education in Australia reflected the influences of a public primary school, a prestigious private high school, and undergraduate and graduate work at Australia's oldest university. His approach to hypnosis was influenced initially by Gordon Hammer and Philip Sutcliffe, and his life generally was influenced by John Anderson, the leader of the Libertarian Society, the intellectual core of a broader group known as the Push. This group reflected in part the thinking of the pre-Socratic Greek philosopher, Heraclitus, who taught that change was the only reality. The article summarizes Perry's work on hypnosis and memory, and his contributions concerning uncancelled hypnotic suggestions, pain and surgery, and imagery and hypnotizability are summarized. PMID- 15115058 TI - The historical role of hypnosis in the theoretical origins of transference. AB - There has been a gradual evolution of the important construct of transference from ancient to modern times. Long before Franz Anton Mesmer, there were philosophers, theorists, and health professionals who emphasized the impact of interpersonal relationships on well-being and illness. While basically conceptualizing animal magnetism as a dynamic physical fluid, Mesmer was also aware of the impact of rapport and affect in the enhancement of magnetic treatment. Later neo-mesmerists, notably Puysegur and Deleuze, built on such theories. That emphasis continued through the 19th century to the time of Freud, whose construct of transference was derived from his personal experience with hypnosis and which has since become an integral part of modern psychodynamic theory and treatment. PMID- 15115059 TI - Four decades of group hypnosis scales: what does item-response theory tell us about what we've been measuring? AB - To overcome problems with previous psychometric approaches to hypnosis scales, the authors applied full-information factor analysis, based on multidimensional item-response theory (IRT), to a 39-year sample of 11,517 records of the Harvard Group Scale of Hypnotic Susceptibility, Form A (HGSHS:A). They also performed a comparable analysis on the standardization sample of the Waterloo-Stanford Group C Scale (WSGC). The HGSHS:A emerges as two-factored, whereas the WSGC more closely approaches unidimensionality. The HGSHS:A factor structure and means show very little change over 4 decades. However, IRT-based item analysis on the HGSHS:A indicates that problems such as "pseudoguessing" on 2 items limit the quality of the item set. The authors propose alternative substantive interpretations of the traits that may underlie the two-factor structure. PMID- 15115060 TI - See clearly: suggestion, hypnosis, attention, and visual acuity. AB - Some reports claim that positive suggestion (e.g., using hypnosis) can significantly improve visual acuity (e.g., in myopes). Based on behavioral, neurocognitive, and ophthalmological findings, the authors provide a critical account to review and challenge some of these data. While acknowledging the relative merits of hypnosis for investigating visual phenomena, an array of arguments converges to propose caveats to the apparent influence suggestion can exert on visual acuity. The authors argue that neither suggestion nor hypnotic phenomena are likely to significantly improve myopic vision and contend that a responsible scientific attitude should carefully outline what hypnosis and suggestion cannot do in addition to what they can. It seems likely that the small apparent influence of suggestion on visual acuity is mediated by changes in attention. The authors outline how attention can affect visual acuity. PMID- 15115061 TI - Reality monitoring in hypnosis: a pilot investigation. AB - In a pilot investigation of reality monitoring in hypnosis, 10 high and 10 low hypnotizable participants were administered a hypnotic suggestion to hallucinate a visual shape on a wall. For half the participants, an image was subtly projected onto the wall at the commencement of the suggestion and then subsequently removed. For the remaining participants, the projected image was initially absent and subsequently projected. Participants completed ratings of belief in the suggestion during hypnosis and also provided subjective reports of the suggestion during a subsequent Experiential Analysis Technique session. High hypnotizable participants who had the projected image introduced at the end of the suggestion provided comparable belief ratings when the image was present and absent. In contrast, highs who had the projected image presented first reported less belief when the image was absent than when it was present. Low hypnotizable participants rated the hallucination more strongly when the image was projected than when it was not projected. These pilot data are discussed in terms of developing a paradigm to objectively index the perceived reality of hypnotically suggested experiences. PMID- 15115062 TI - The effect of postnatal depression on mother-infant interaction, infant response to the Still-face perturbation, and performance on an Instrumental Learning task. AB - A representative community sample of primiparous depressed women and a nondepressed control group were assessed while in interaction with their infants at 2 months postpartum. At 3 months, infants were assessed on the Still-face perturbation of face to face interaction, and a subsample completed an Instrumental Learning paradigm. Compared to nondepressed women, depressed mothers' interactions were both less contingent and less affectively attuned to infant behavior. Postnatal depression did not adversely affect the infant's performance in either the Still-face perturbation or the Instrumental Learning assessment. Maternal responsiveness in interactions at 2 months predicted the infant's performance in the Instrumental Learning assessment but not in the Still face perturbation. The implications of these findings for theories of infant cognitive and emotional development are discussed. PMID- 15115063 TI - Predicting the development of infant emotionality from maternal characteristics. AB - Few studies have examined the associations between environmental conditions and developing infant emotionality or the differential susceptibility to those conditions. The present longitudinal study aims to make a contribution to close that gap. We analyzed whether positive emotionality, negative emotionality/irritability, and withdrawal/fear at the end of the first year of life are predictable from preceding caregiver's depression/anxiety, social support, and sensitivity in the interaction with the infant while controlling for antecedent states of emotionality. Furthermore, the question of whether associations between maternal characteristics and subsequent fear are stronger in the subgroup of infants high in irritability as opposed to those who are low in irritability was investigated. Subjects were 101 healthy firstborn infants and their primary caregivers. Assessments were conducted at infant ages of 4, 8, and 12 months. Depression, anxiety, and the social support of the caregiver were assessed by questionnaire. Sensitivity in the caregiver-infant interaction was assessed by behavior observations within the scope of home visits. Temperament characteristics were observed in standardized laboratory episodes. Whereas negative emotionality and withdrawal/fear were significantly predictable from the maternal characteristics, no predictability could be shown for developing positive emotionality. There were indications of a stronger association between the maternal characteristics and developing withdrawal/fear in irritable infants. PMID- 15115064 TI - Affect dysregulation in the mother-child relationship in the toddler years: antecedents and consequences. AB - The purpose of this study was to examine child, maternal, and family antecedents of children's early affect dysregulation within the mother-child relationship and later cognitive and socioemotional correlates of affect dysregulation. Children's affect dysregulation at 24 and 36 months was defined in the context of mother child interactions in semistructured play and toy cleanup. Dyads were classified as dysregulated at each age based on high negative affect. Affect dysregulation was associated with less maternal sensitivity and stimulation, more maternal depressive symptoms, and lower family income over the first 36 months of life. Children with early negative mood, lower Bayley Mental Development Index scores and insecure-avoidant (15 months) or insecure-resistant attachment classifications (36 months) were more likely to be in an affect-dysregulated group. Controlling for family and child variables, affect-dysregulated children had more problematic cognitive, social, and behavioral outcomes at 54 months, kindergarten, and first grade. The findings are discussed in terms of the early role played by parents in assisting children with affect regulation, the reciprocal nature of parent-child interactions, and the contribution of affect regulation to children's later cognitive, social, and behavioral competence. PMID- 15115065 TI - An integrative approach to the neurophysiological substrates of social withdrawal and aggression. AB - An integrative model of the neurophysiology of aggression and social withdrawal is proposed. A detailed overview of the limbic-hypothalamic-pituitary-adrenal (LHPA) axis is presented first, because we consider it to be a critical system that interacts with a variety of physiological processes to modulate affect related behaviors. This detailed analysis of the LHPA axis is then used to clarify the research literature that links aggression and social withdrawal to LHPA functioning. We then review the role of amygdala and prefrontal cortex functioning in modulating aggression and social withdrawal. Particular attention is paid to how the amygdala and the prefrontal cortex interact with the LHPA system and the environment to produce specific behavioral tendencies throughout development. A brief overview of the implied methodological and theoretical model is provided. We explain how a detailed understanding of specific physiological processes is essential in order to develop appropriate research protocols. In addition, we suggest that future research should focus on the mapping of distinct integrative biosocial profiles that are related to specific behaviors during different developmental stages. PMID- 15115066 TI - Emotion-related abilities and depressive symptoms in Latina mothers and their children. AB - This study examined the abilities of 40 Latina mothers and their 6- to 11-year old children (20 girls, 20 boys) to recognize and produce emotion expressions and how these abilities differed as a function of maternal depressive symptoms. The results indicated that depressively symptomatic mothers were less accurate at recognizing basic emotions (e.g., happy, sad, etc.) and some mixed emotions (e.g., scared/ok combinations) than nonsymptomatic mothers, but there were no group differences for emotion production. In contrast, children of symptomatic mothers posed fewer recognizable sad expressions than their peers. Error pattern analyses also revealed that children of symptomatic mothers were more likely to mistakenly recognize happiness and to avoid posing sadness (across all basic emotions). Children's ability to pose emotions was related to their mothers' emotion production, and this was not moderated by maternal depressive symptoms. The discussion focuses on the possible interpersonal consequences of these biases and deficits in the emotion-related abilities of symptomatic mothers and their children and on the need to conduct research on the familial and cultural processes that might underlie these findings. PMID- 15115067 TI - The impact of high neuroticism in parents on children's psychosocial functioning in a population at high risk for major affective disorder: a family-environmental pathway of intergenerational risk. AB - Behavioral genetic studies indicate that nongenetic factors play a role in the development of bipolar and major depressive disorders. The trait of neuroticism is common among individuals with major affective disorders. We hypothesized that high neuroticism among parents affects the family environment and parenting practices and thereby increases the risk of psychosocial problems among offspring. This hypothesis is tested in a sample of participants at high and low risk for major affective disorders, which contained parents with bipolar disorder (55), major depression (21), or no mental disorder (148) and their 146 children between 4 and 14 years of age. Parents with high neuroticism scores were characterized by low psychosocial functioning, poor parenting, more dependent stressful life events, and the use of more emotion-focused and less task-oriented coping skills. High neuroticism in parents was associated with internalizing and externalizing problems among the children, as assessed by parent and teacher ratings on the Child Behavior Checklist and clinician ratings. The results suggest that high neuroticism in parents with major affective disorders is associated with inadequate parenting practices and the creation of a stressful family environment, which are subsequently related to psychosocial problems among the offspring. PMID- 15115069 TI - The expression and understanding of jealousy in children with autism. AB - We investigated the expression and understanding of jealousy in 16 high functioning children with autism and 17 typically developing children matched for IQ, chronological age, gender, and maternal education. We examined the expression of jealousy via children's behaviors, verbalizations, and affects demonstrated during two jealousy-provoking triadic scenarios (drawing and playing) enacted among the child in the experimental group (autism or typical), that child's main caregiver (mostly mothers), and a familiar peer or sibling. The two scenarios corresponded with the two types of jealousy described in past studies: social comparison jealousy (drawing scenario) and social-relational jealousy (playing scenario). To tap children's understanding, we asked them to identify jealousy from a picture, to provide examples of times they felt jealous, and to offer suggestions for coping with jealousy. The main results revealed that children with autism expressed jealousy in situations similar to their typical age mates but manifested it in different behaviors. Moreover, children with autism revealed a less coherent understanding of the feeling. We discuss the meaning of the gap between demonstrating and understanding jealousy in light of the two central theoretical views conceptualizing the core emotional deficit in children with autism. PMID- 15115068 TI - The relationship of theory of mind and executive functions to symptom type and severity in children with autism. AB - Although neurocognitive impairments in theory of mind and in executive functions have both been hypothesized to play a causal role in autism, there has been little research investigating the explanatory power of these impairments with regard to autistic symptomatology. The present study examined the degree to which individual differences in theory of mind and executive functions could explain variations in the severity of autism symptoms. Participants included 31 verbal, school-aged children with autism who were administered a battery of tests assessing the understanding of mental states (knowledge and false belief) and executive control skills (working memory, combined working memory and inhibitory control, and planning) and who were behaviorally evaluated for autism severity in the three core symptom domains. Whereas theory of mind and executive control abilities explained the significant variance beyond that accounted for by language level in communication symptoms, neither explained the significant variance in reciprocal social interaction or repetitive behaviors symptoms. These findings are discussed in terms of a proposed distinction between higher level, cognitive-linguistic aspects of theory of mind and related executive control skills, and more fundamental social-perceptual processes involved in the apprehension of mental state information conveyed through eyes, faces, and voices, which may be more closely linked to autistic deficits in social reciprocity. PMID- 15115070 TI - Gender differences in childhood onset conduct disorder. AB - The present study tested several predictions related to the theory that girls with conduct disorder (CD) follow a single delayed onset pathway. The following hypotheses were tested among a high-risk, stratified random sample of youth who used public services during a 6-month period: boys will be more likely to have childhood onset CD (COCD) than girls; girls with COCD will be less common than girls with adolescent onset CD (AOCD); COCD girls. AOCD girls, and COCD boys will be similar in terms of risk profiles and AOCD boys will have a less severe risk profile than COCD girls; and risk factors that differentiate between COCD and AOCD among males will not differentiate between COCD and AOCD among females. Among those youth who met the criteria for CD, males were significantly more likely to have COCD than females. However, close to half of females with CD met the criteria for the COCD subtype. Of the seven risk factors that were examined, girls with COCD scored higher than COCD boys on three factors and higher than AOCD girls on four factors; however, only one significant difference was found between AOCD girls and COCD boys. Finally, risk factors appeared to differentiate between COCD and AOCD groups in a similar way among males and females. These findings suggest that COCD is not rare among females in public service sectors, COCD girls can be distinguished from AOCD girls in terms of risk factors, and risk factors for COCD among males are also relevant to females. PMID- 15115071 TI - Early adolescent through young adult alcohol and marijuana use trajectories: early predictors, young adult outcomes, and predictive utility. AB - The present study takes a developmental approach to subgrouping and examines the trajectories of substance use from early adolescence through young adulthood among a community sample of 481 individuals. The patterns of use were examined, subgroups were identified separately for men and women and for alcohol and marijuana, and psychosocial predictors and psychopathology outcomes that differentiated the groups were identified. The results revealed three substantially overlapping subgroups for both alcohol and marijuana: early onset, late onset, and nonuser. Although the general patterns of which dependent variables were related to group were similar for alcohol and marijuana, a closer examination revealed important subgroup differences. For alcohol use, the early onset group was more dysfunctional in terms of predictors and outcomes whereas the late-onset and nonuser groups were better adjusted. In contrast, for marijuana, the early- and late-onset groups were both more dysfunctional than the nonuser group. In a final analysis, we examined the predictive utility of our developmental approach to subgrouping compared to a traditional, static approach. PMID- 15115072 TI - Factors associated with the unresolved classification of the Adult Attachment Interview in women who have suffered stillbirth. AB - The "unresolved" state of mind with respect to loss or trauma as assessed in the Adult Attachment Interview is common in clinical and forensic groups, as well as in mothers whose infants are classified as disorganized in their attachment relationship to them. However, questions remain about what the unresolved state represents and what factors predict the unresolved state. This case controlled study reports on 64 women who had suffered stillbirth and who were pregnant with their next child. The study explores attachment, psychiatric, and social factors associated with the unresolved state or higher unresolved scores with respect to stillbirth. Women who had experienced stillbirth were more likely to be unresolved than control women. Although a similar number of stillbirth and control women had experienced childhood trauma, only women who had experienced stillbirth were unresolved with respect to this trauma, suggesting the unresolved state may be evoked or reevoked by subsequent traumatic loss. Higher unresolved scores in relation to stillbirth were predicted by childhood trauma, poor support from family after the loss, and having a funeral for the infant. The results are discussed in terms of the woman's sense of being causal in the loss. PMID- 15115073 TI - Protective immunity induced with 23 kDa membrane protein dna vaccine of Schistosoma japonicum Chinese strain in infected C57BL/6 mice. AB - A 23 kDa membrane protein DNA vaccine for Schistosoma japonicum Chinese strain was developed and tested for its protective efficacy and immune responses in infected C57BL/6 mice. The cDNA encoding SjC23 amplified from pUC19-SjC23 were subcloned into an eukaryotic expression vector (pcDNA3.1). Forty-eight female C57BL/6 mice were divided into three groups. Each mouse of group A (control group) was immunized intramuscularly (i.m.) with 100 microg of pcDNA3.1; of group B (SjC23 group) was immunized (i.m.) with 100 microg of pcDNA3.1-SjC23; of group C (SjC23+IL-12) was immunized (i.m.) with a mixture of 100 microg of pcDNA3.1 SjC23, 100 microg of pcDNA3.1-p35 and 100 microg of pcDNA-p40. These were followed by two boosts of the same DNA once every two weeks. All mice were challenged with 45 cercariae of Schistosoma japonicum Chinese strain at week 8, and were killed and perfused at week 14. The numbers of recovered worms and hepatic eggs were counted. The expression of SjC23 and p35, p40 in muscle tissue was determined by immunohistochemical method. By culture of spleen cells, the production of IL-2, IL-4, IL-10 and IFN-gamma with the stimulation of specific antigen of the recombinant hydrophilic domain of SjC23 (rSjC23-HD) was determined after the last immunization (before challenge). Sera were collected from each group before immunization and two weeks before and after challenge. Anti-SjC23 antibodies were tested by Western blot. The results showed that SjC23 and p35, p40 of mouse IL-12 were expressed on the membrane and in the plasma of the muscle cells of immunized C57BL/6 mice. A rise of IL-2 and IFN-gamma in the SjC23 group and SjC23+IL-12 group was observed; No changes were found in IL-4 and IL-10. Detection of anti-SjC23 antibody with Western blot showed that after the third immunization (before challenge) all the serum samples from the control group were negative; 8 of 10 sera from the SjC23 group and 9 of 10 sera from the SjC23+IL-12 group were positive. The worm reduction rates in the SjC23 group and SjC23+IL-12 group were 26.9% and 35.4% respectively; the liver eggs reduction rates were 22.2% and 28.4%, respectively in comparison to the control group. This indicates that the pcDNA3.1-SjC23 DNA vaccine can induce partial protection against Schistosoma japonicum infection in C57BL/6 mice. PMID- 15115074 TI - Immunoblot analysis of Schistosoma japonicum egg antigens with sera from patients with acute and chronic schistosomiasis japonica. AB - Humoral immune responses of IgG, IgM, IgA, IgE and IgG subclass antibodies to Schistosoma japonicum egg antigens were determined by immunoblotting with serum samples from individuals in China with acute (n=24) or chronic (n=35) schistosomiasis. In general, IgM, IgA, and IgE in sera from acute patients exhibited strong binding to antigens but binding was much weaker in chronic cases. Reaction of IgG4 of chronic cases was stronger than that of IgG4 of acute cases. The recognition profile of each antibody isotype in sera was analyzed for 11 major antigen molecules (antigens with apparent molecular weights of 82, 76, 61, 57, 53, 46, 40, 32, 27, 10 and less than 6.5 kDa). Except for the 10 kDa molecule, they were well-recognized by IgA and IgE in sera of acute cases. In other combinations of antibody class and clinical phase, recognition patterns against these molecules differed among individuals. Notably, the 10 kDa molecule was specifically recognized by total IgG and IgG4 in sera from most of the chronic patients, but in sera from only one acute case. This result suggests that the 10 kDa molecule is one of the major target antigens of IgG4 and may be useful as a marker antigen to characterize the clinical phases of S. japonicum infection. PMID- 15115075 TI - Egg production capacity of one-pair worms of Schistosoma japonicum in albino mice. AB - In the present study, a series of procedures of egg count were carried out to determine the egg production capacity in 21 ICR mice each infected with one-pair of Schistosoma japonicum. The egg count began from the first day, they were detected in the feces, which was based on a stool collection over 24 hours, twice a week. Each female S. japonicum produced an average of 2,198 eggs/day during the study period of 99 days after infection (ranged 61-147 days). Fourty-seven percent of the eggs were in the feces and 53% in tissues (45% in large intestine, 31% in small intestine, 23% in liver, 0.4% in pancreas, 0.2% in lungs, 0.1% in spleen, 0.1% in lymph nodes, 0.06% in stomach and 0.05% in heart, kidney, diaphragm and brain). PMID- 15115076 TI - Immunodiagnosis of human fascioliasis using an antigen of Fasciola gigantica adult worm with the molecular mass of 27 kDa by a dot-ELISA. AB - Immunodominant antigens of an approximate molecular mass of 27 kDa (FG 27) were obtained from an excretory-secretory product of adult Fasciola gigantica by a simple continuous-elution method. A dot-ELISA using the FG 27 antigen was developed for the detection of specific antibodies from patients infected with F. gigantica. Control sera were obtained from patients with other parasitic infections and healthy volunteers. The accuracy, sensitivity, specificity, and positive and negative predictive values were 98.2%, 100%, 97.4%, 76.9% and 100%, respectively. This dot-ELISA is a specific, sensitive and easy to perform method for the rapid diagnosis of fascioliasis, particularly when more complex laboratory tests are unavailable. PMID- 15115077 TI - Recovery and growth of Haplorchis taichui (Trematoda: Heterophyidae) in chicks. AB - An experimental study was performed to observe the recovery and growth of a minute intestinal fluke, Haplorchis taichui in chicks (Gallus gallus domesticus). Metacercariae of H. taichui were isolated from Jullien's mud carp, Henicorhynchus siamensis, which were collected in the Chiang Mai Province, Thailand. Two hundred metacercariae were orally force-fed to each chick. The intestine of the chicks were examined from day 1 to day 54 post-infection (PI). The incidence of infection was 84.9% (28/33) and the mean intensity was 19.9 (656/33), with the range 0-59. The worm recovery rate was the highest at day 11 PI (29.5%). On day 3 PI, mature adult worms were recovered and 1-200 eggs were observed in the uterus of the worms. The worms grew rapidly in the chicks and the genital organs were fully developed in 14 days. This parasite can survive in chicks up to day 48 PI. It is concluded that they are a suitable definitive host for infection with H. taichui. PMID- 15115078 TI - Recombinant expression of Toxocara canis excretory-secretory antigen TES-120 in Escherichia coli. AB - The gene encoding the excretory-secretory antigen TES-120 of dog ascarid worm Toxocara canis was cloned into the bacterium Escherichia coli. The specificity of the recombinant TES-120 antigen produced by the bacterium was investigated. A total of 45 human serum samples from patients infected with differenthelminthes and protozoa, including 8 cases of toxocariasis, were tested against the recombinant antigens in immunoblot assays. The results from the assays revealed that the recombinant TES-120 antigen reacted with sera from toxocariasis patients only. This highly specific recombinant TES-120 antigen can potentially be used for the development of an inexpensive serodiagnostic assay for human toxocariasis. PMID- 15115079 TI - Polypeptides associated with in vitro cyst formation of Blastocystis hominis. AB - The objective of this study was to characterize the polypeptides associated with cysts of Blastocystis hominis. This form is believed to be infective and plays a role in parasite resistance to anti-B. hominis drugs currently used for treatment of Blastocystis associated diarrhea. Cysts were induced through in vitro culture of the parasite in complete medium supplemented with bacterial extract with trypticase, metronidazole or doxycycline. SDS-PAGE analysis showed almost similar polypeptide patterns of parasite extracts obtained from in vitro cultured parasites before and after exposure with the three supplements. Polypeptide bands at 76, 58.5, 48, 45, 40, 38, 32, 25 and 22 kDa were constantly seen in all antigenic preparations and no specific cyst-associated polypeptide was present. However, on immunoblot analysis, 3 out of 16 blastocystosis human sera identified a cyst-associated polypeptide at 60 kDa in all parasite extracts prepared from cultures with the three supplements. In addition, there were associated morphological changes detected in these parasites stained with acridine orange and observed under fluorescence microscopy. Metronidazole induced cyst forms (reddish cells) as early as 12 hours post-exposure; more cyst production (with stronger immunoblot bands) occurred after 24 hours exposure. However, cysts rupture with release and destruction of B. hominis daughters cells occurred after 48 hours exposure. Doxycycline induced less cyst-like forms at 24 hours (weaker 60 kDa band) and less destruction of the cysts (60 kDa band still present at 72 hours post exposure). Bacterial extract and trypticase also induced cysts at 12 hours with increasing numbers up to 72 hours exposure (corresponding increase in intensity of 60 kDa band from samples harvested at 12 to 72 hours post exposure) without any sign of deleterious effect on the parasite. PMID- 15115080 TI - Toxoplasma gondii antibody in Thai cats and their owners. AB - Humans are thought to acquire Toxoplasma infection by three major routes: ingesting food and water contaminated with oocysts from cat excreta, consumption of under-cooked infected meat, and transplacental transfer. Congenital clinical toxoplasmosis in the newborn indicating definite transplacental transmission had been reported in Thailand, whilst studies concerning infection due to the other two routes were inconclusive. Since the way domestic cats live and eat and also the eating behavior of Thai people differ from those in the West, we conducted a sero-epidemiological study of T. gondii in cats and their owners in Bangkok metropolitan area. Among 327 humans, the prevalence of Toxoplasma antibody was 6.4% and in 315 cats it was 7.3%. These relatively low prevalence rates may result from the predominantly well-cooked fish and rice diet of stray cats, which congregate in temples where they are fed. Toxoplasma antibody seropositive was associated with living in close proximity to seropositivity cats [OR (95% CI) = 5.43 (1.28-23.04); p=0.01]. Risks were increased in and around temples, particularly if courtyards were of earth or grass, suggesting ground temperature was an important determinant of oocyst survival. PMID- 15115081 TI - Purification and partial characterization of PfHRP-II protein of Plasmodium falciparum. AB - The human malarial parasite Plasmodium falciparum secretes various intra-and extra-cellular proteins during its asexual life cycle in human RBC. Histidine rich protein-II (HRP-II) is one of the most prominent proteins, found to be secreted by P. falciparum throughout the asexual cycle with the peak during mature schizont stage of the parasite development in human IRBC. The high histidine content (35% of the total amino acids in protein) of this protein suggested the potential to bind divalent metal ions. We have demonstrated by metal chelate chromatography, an extraordinary capacity of HRP-II to bind nickel ions (Ni++) and employed this characteristic to purify the extra-cellular HRP-II protein secreted by P. falciparum from culture supernatant. The identity of the purified protein was verified by the relative molecular weight on SDS-PAGE, by reacting with polyclonal antibodies directed against it using Western blot technique. PMID- 15115082 TI - Effect of dihydroartemisinin on the antioxidant capacity of P. falciparum infected erythrocytes. AB - Many lines of evidence reveal that artemisinin, an antimalarial containing endoperoxide, generates free radicals to kill malaria parasites. The present study re-evaluated the antioxidants of P. falciparum-infected erythrocytes in the absence and presence of 0.25, 0.5 and 1.0 ng/ml of dihydroartemisinin (DHA), the active metabolite of artemisinin. The ratio of reduced to oxidized glutathione (GSH/GSSG) and activities of superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) were determined. The data indicated that malaria infection induced oxidative stress in erythrocytes that resulted in a significant lower GSH in parasitized cells compared to the non-parasitized. DHA showed no effect on the antioxidant levels of non-parasitized erythrocytes treated under similar conditions as P. falciparum-infected erythrocytes. However, significantly lower GSH as well as catalase and GPx activities in parasitized cells were seen at drug concentrations of 0.5 and 1.0 ng/ml (p < 0.05). GSH is the most sensitive indicator of oxidative stress in malaria-infected erythrocytes both in the absence and in the presence of DHA. Parasite GPx might play a more important role than catalase in the elimination of peroxide. Parasite viabilities in the presence of DHA were analyzed simultaneously and were affected to a greater extent than the antioxidant levels. The present observation showed that although DHA killed malaria parasites by generating free radicals from the endoperoxide bridge causing the reduction of antioxidants, but the depletion of parasite antioxidants is not a prerequisite for the parasite death. PMID- 15115083 TI - Detection of Babesia bovis in cattle by PCR-ELISA. AB - We established a new highly sensitive method, PCR-ELISA, for the dectection of Babesia bovis in cattle for farms in Thailand. The detection of around 2.4 x 10( 8)% parasitemia (equivalent to 1 infected erythrocyte per 2 ml) was achieved by PCR amplification followed by the ELISA detection of a biotin tagged gene. When comparing the sensitivity of PCR-ELISA with the microscopic method, our PCR-ELISA method is more sensitive than thin blood smears by at least 1,000 times. The established PCR-ELISA also showed high specificity to B. bovis with no cross reaction to other endemic parasites except for A. marginale. Regarding the detection threshold for B. bovis, the PCR-ELISA method could detect parasites inoculated into splenectomized calves at least 1 week earlier than the thin blood microscopic method. The PCR-ELISA method is a valuable screening technique for B. bovis and applicable for the routine detection of carrier states and automated analysis. PMID- 15115084 TI - Wuchereria bancrofti antigenemia clearance among Myanmar migrants after biannual mass treatments with diethylcarbamazine, 300 mg oral-dose FILADEC tablet, in Southern Thailand. AB - Using qualitative ICT Filariasis and quantitative Og4C3 ELISA, we assessed a long term macrofilaricidal effect of two-year biannual mass treatments with a 300 mg oral-dose FILADEC tablet, a reformulation of 6 mg/kg diethylcarbamazine (DEC), on clearance of the Wuchereria bancrofti adult worm circulating filarial antigens (CFA) in Myanmar migrants, at risk of emergence of imported bancroftian filariasis in Southern Thailand. Of the 34 antigenemic Myanmar index cases of varying initial CFA levels, who were initially screened out with the ICT Filariasis, 13 index cases were follow-up treated and monitored at the DEC post treatments, 6, 12, and 18 months. At the 18-month post treatment, residual antigenemias (%) in 4 of 5 index cases (group 1) with high antigen titers (99.7 181.6 x 10(3) AU/ml) were 54.44%, 33.58%, 27.43%, and 9.97%. Significant decreases of the CFA levels in only 3 out of 5 index cases were affected by the response to DEC treatments (p < 0.007). The treatment effects on clearance of the CFA in 8 index cases (group II) with low antigen titers (15.4-37.2 x 10(3) AU/ml) were shown for at least 6 months post DEC treatment and hence had 100% efficacy in the first 6 months of the first year of year round treatment. Group I, was more likely to show an increase of the DEC efficacy after the first 6 months of the second year round treatment, but there was no statistically significant difference (p = 0.063). We reemphasized that, for use in the national program to eliminate lymphatic filariasis (PELF) in Thailand, such a DEC regimen had a macrofilaricidal effect on antigenemia clearance, and confirmed its value in evaluating response to the treatment and monitoring the long-term efficacy of the DEC regimen in W. bancrofti adult worm burden reductions in Myanmar migrants on a wide scale. PMID- 15115085 TI - Comparison of two IgG4 assay formats (ELISA and rapid dipstick test) for detection of brugian filariasis. AB - Brugia malayi infection is endemic in several Asian countries. Filaria-specific IgG4 antibody detection based on BmR1 recombinant antigen has been shown to be sensitive and specific for the diagnosis of brugian filariasis. Two formats of the test has been reported ie indirect ELISA (BE) and rapid dipstick test (BR). Since different test formats use different amounts of sample and reagents which may affect its sensitivity and specificity, this study was performed to compare these two test formats in the detection of B. malayi. A total of 264 blinded serum samples from India and Malaysia were employed. Group 1 comprised 164 samples from actively infected individuals and group 2 comprised 100 samples from filaria non-endemic areas. Sensitivity was 96.3% (158/164) and 90.8% (149/164) for rapid test and ELISA respectively; chi-square p=0.00. Both test formats demonstrated 100% specificity. Therefore the rapid test format was equally specific but more sensitive than the ELISA format. The ELISA format would be able to demonstrate decline in IgG4 titer post-treatment while the rapid test would be very useful for screening and diagnosis in the field. PMID- 15115086 TI - Molecular variation and phylogeny of the Anopheles minimus complex (Diptera: Culicidae) inhabiting Southeast Asian countries, based on ribosomal DNA internal transcribed spacers, ITS1 and 2, and the 28S D3 sequences. AB - Anopheles minimus (Theobald) is one of the most important vectors of human malaria in Southeast Asia. Morphological studies now have revealed five sibling species as its complex, designated as species A to E. The present study investigated the genetic divergence among An. minimus populations from four countries (Japan, China, Thailand and Indonesia), based on the DNA sequences data of the D3 (the third domain of the 28S ribosomal gene) and ITS2 (the second internal transcribed spacer of the ribosomal gene) is reported. The D3 and ITS2 phylogenetic trees, and the electrophoretic profile of ITS1 (the first internal transcribed spacer of the ribosomal gene) indicated that our An. minimus populations are comprised of three groups: the Japanese population as group I, the population from Guangxi Province of China (GX population) as group II, and others, as group III. The results showed the morphological similarity of group III and GX with the species complex A and B, respectively. It is interesting that both two species A (YN population) and species B (GX) occur in China, and that both species, An. minimus species A (LB-95 population) and the closer population An. flavirostris (Ludlow) (LB-00 population) appeared to be present on the Lombok Island of Indonesia, although in far separated localities. Moreover, this molecular evidence confirms the previous suggestion that the population from the Ishigaki Island of Japan should be classified as a new genetic status species E. PMID- 15115087 TI - Determination of dengue virus serotypes in Thailand using PCR based method. AB - A reverse transcriptase-polymerase chain reaction (RT-PCR) and a single-tube multiplex PCR assay was modified for typing of dengue virus in different geographical areas of Thailand during 2000-2001. A set of primers (D1 and D2) was used to generate the RT-PCR product of 511 bp in size which subsequently underwent a single-tube multiplex PCR amplification using the highly specific primers for each of the dengue virus serotypes (D1, TS1, TS2, TS3 and DEN4). The PCR products of 482, 119, 290 and 392 bp in size were generated for dengue virus serotypes 1, 2, 3, and 4, respectively. Each set of specific primers showed no amplification of non-specific and non-target PCR products from human genomic DNA. The method was applied for investigation of 637 human blood samples in Thailand during 2000-2001 and found that 71, 43, 28, and 43 patients were classified as having a single infection with serotypes 1, 2, 3, and 4, respectively. Multiple infections with two or more dengue virus serotypes were also detected. PMID- 15115089 TI - Perinatal dengue infection: a case report and review of literature. AB - A case of vertical transmission of dengue infection in the perinatal period is reported. The mother, a term pregnancy, had acute dengue the day before admission. The infant was born at term and developed fever on the fifth day of life which lasted for 5 days. No bleeding or plasma leakage was detected during the course of fever in infant or mother. A liver function test showed elevated SGOT and SGPT in the infant. The infant developed a convalescent rash on day 5 of the fever. The diagnosis of secondary dengue hemorrhagic fever in the mother was confirmed by serology and primary dengue infection in the infant was confirmed by serology and serotyped as dengue type 2 by PCR. The clinical course and management of mothers and infants with perinatal dengue infection are reviewed. PMID- 15115088 TI - Temephos resistance in two forms of Aedes aegypti and its significance for the resistance mechanism. AB - Aedes aegypti, at the larval stage, has been subjected to the temephos selection in laboratory. The level of temephos resistance was detected in a microplate by biochemical assay using WHO bioassay technique. The major enzyme-based resistance mechanisms involved in temephos resistance include elevated nonspecific esterase, oxidase and insensitive acetylcholinesterase. After 19 generations of temephos selection, the selected group showed resistance ratios of 4.64 and 16.92, when compared with a non-selected group and the WHO susceptible strain, respectively. The two seperated forms, type form and the pale form of Ae. aegypti showed low levels of resistance to temephos after 19 generations of selection, with resistance ratios of 4.82 and 4.07 for the type form and the pale form, respectively; when compared with the non-selected strain, 17.58 and 14.84, when compared with the WHO susceptible strain. This showed that the type form could develop higher level resistance than the pale form. The esterase inhibitor (S,S,S tributyl phosphorotrithioate, DEF) or synergist implicated detoxifying esterase in all the temephos selected groups and the presence of elevated esterase were confirmed by biochemical assay. There were significant differences in elevated esterase activity between the temephos selected groups and the non-selected group. However no significant difference between the type form and the pale form was found. Besides the elevated esterase, there was no change in monooxygenase activity and no evidence of insensitive acetylcholinesterease for all temephos selected groups. These results suggest that temephos resistance could be developed in Ae. aegypti under selection pressure and that the main mechanism is based only on esterase detoxification. PMID- 15115090 TI - Parturient and perinatal dengue hemorrhagic fever. AB - A Thai woman in the shock stage of dengue hemorrhagic fever delivered a healthy infant by normal delivery. She had high, prolonged fever for five days, hepatomegaly, thrombocytopenia and a right pleural effusion. The serology performed later established a diagnosis of secondary dengue infection. She had no serious complications except for postpartum anemia. The newborn became febrile at the 48th hour of life. He had fever for 2 days, hepatomegaly, thrombocytopenia and a right pleural effusion. Dengue virus type 1 was detected from his serum by polymerase chain reaction (PCR). Although he developed prolonged, marked thrombocytopenia (12 days), his illness was uncomplicated. He recovered uneventfully and was discharged with his mother. This report was one of dengue vertical transmission with dengue hemorrhagic fever in humans. PMID- 15115091 TI - Vertical transmission of dengue: first case report from Bangladesh. AB - We report a case of vertical transmission of dengue infection in a new born from Bangladesh. The mother was primigravida with an uneventful antenatal period except the fever she developed at 37 weeks of gestation. She underwent cesarean section for decreased fetal movement. The fever lasted for 7 days and was associated with intense bodyache, and a positive tourniquet test. She did not experience any hemorrhages including from the surgical wound. Her platelet count fell at its lowest to 100,000/mm3. She had bilateral mild pleural effusions and positive anti-dengue antibodies (both IgG and IgM). She received symptomatic treatment. The baby was deeply meconium stained and was resuscitated by a pediatrician. His postnatal period went well until day-3 of life when he developed fever and respiratory distress. There were no signs of meconiun aspiration syndrome (MAS) and the septic work up was negative. The platelets count fell to its lowest of 40,000/mm3 on day-3 and day-5 of his illness. Anti dengue antibodies (both IgG and IgM) were positive with a nearly four-fold rise of IgM antibodies in the convalescent sera. The boy was treated with platelet transfusions and was discharged on day-6 after becoming ill. This report emphasizes that in a dengue epidemic or when dengue is endemic (which Bangladesh has experienced recently), a pregnant woman with fever, myalgia and/or bleeding manifestations should raise a high suspicion that the baby may develop the disease, and both the mother and baby should be closely followed-up. Viral isolation could not be done due to the unavailability of the test. PMID- 15115092 TI - Factors associated with tuberculin skin test reactivity among HIV-infected people in Bangkok. AB - INH preventive therapy (IPT) has been shown in several randomized controlled trials to reduce the risk of developing active TB in tuberculin skin test (TST) or purified protein derivative (PPD) positive HIV infected individuals. Detection of latent tuberculosis by TST and determination of factors associated with the PPD positivity in HIV-infected persons are important for the targeting of chemoprophylaxis. Six hundred asymptomatic and early symptomatic HIV-infected subjects attending the AIDS Clinic of the Chulalongkorn University Hospital, Bangkok, Thailand were enrolled in two randomized clinical trials of chemoprophylaxis against TB from December 1994 to December 1996. The availability of baseline characteristics, including TST reactivity, among these participants enabled a cross-sectional analysis of factors associated with PPD positivity. The results showed that 117 (19.5%) were PPD positive and 483 (80.5%) were PPD negative with ages 18-65 years (median 29 years). HIV exposure category was 46.2%, 34.5%, and 6.7% for heterosexual contact, commercial sex work, and homosexual and bisexual male contact respectively. The median CD4 cell count was 315/mm3 (range, 5-1,074/mm3). HIV exposure category and CD4 cell count were significantly associated with PPD status. Homosexual/bisexual contact had 3 times higher risk of PPD positivity than heterosexual contact (adjusted OR=2.9; 95% CI, 1.4-6.1) and risk of PPD positivity was higher among patients with CD4 cell counts of 200-500/ mm3 (adjusted OR=1.8; 95% CI, 1.0-3.1) and above 500/mm3 (adjusted OR=3.4; 95% CI, 1.7-6.7) when compared to patients with CD4 cell counts of less than 200/mm3. The HIV-infected persons in Bangkok with homosexual/bisexual contact are at higher risk for latent TB. Population-based tuberculin screening without accompanying HIV testing cannot be used to estimate the prevalence of actual latent TB in a population where HIV infection is widespread, such as in Thailand. PMID- 15115093 TI - Searching for virulence Burkholderia pseudomallei genes by immunoscreening the lambda ZAPII expressed genomic library. AB - The lambdaZAP II expressed genomic library of B. pseudomallei was screened with pooled melioidosis serum preabsorbed with E. coli host cell. The positive clones were detected by using protein A-CDP-star chemiluminescence. All of 14 positive clones reacted with only the pooled absorbed melioidosis serum and not the pooled absorbed normal serum when tested with the plaque dot blot analysis. The expressed genes were detected by using a combination of immunoscreening, bioinformatics and molecular biology. At least six in vivo expressed genes were identified by this approach. Two were well known virulent genes, gmhA (a capsule biosynthetic gene) and bipD (type III secretion protein gene). Another two were genes coded for conserved hypothetical protein. The last two isolated genes were groEL (a chaperonine protein gene), and a gene encoding transmembrane protein. PMID- 15115094 TI - Leptospirosis in northern India: a clinical and serological study. AB - A total of 400 serum samples collected from patients, clinically suspected of leptospirosis, were evaluated for antibodies by LEPTO dipstick and microscopic agglutination test (MAT). Twenty of these patients (5%) had serological evidence of leptospirosis. Leptospira interrogans serovars Autumnalis and Icterohaemorrhagiae, Canicola and Javanica were serogroups recorded serologically. Fever and jaundice were the most common clinical presentations. Male preponderance was seen in the leptospirosis cases. Outdoor activities, agricultural activities, contact with animals were significantly associated with seropositivity for Leptospira. This study highlights that leptospirosis is a significant health problem in northern India, though grossly under reported due to the absence of routine laboratory diagnostic facilities for this disease. PMID- 15115095 TI - Epidemiological studies on host animals of scrub typhus of the autumn-winter type in Shandong Province, China. AB - In order to elucidate the host animals of scrub typhus in Shandong Province, epidemiological studies on host rodents of the autumn-winter type scrub typhus were carried out from 1995 to 2002 at four localities in the Shandong Province. Based upon ecological observations of the composition, seasonal fluctuation of animal hosts, isolation of Orientia tsutsugamushi, detection and identification of serotypes of antibodies to O. tsutsugamushi were conducted. Two thousand eight hundred and eighty-four rodents and insectivores were captured, including 2,055 Apodemus agrarius (71.26%), 408 Cricetulus triton (14.15%), 64 C. barabensis (2.22%), 12 Crocidura suaveolens (0.42%), 313 Rattus (R.) norvegicus (10.85%), 32 Mus (M.) musculus (1.11%). A. agrarius was predominant in the field and the seasonal fluctuation was correlated significantly to that of scrub typhus (r=0.810, p<0.005). R. norvegicus was predominant indoors. The average capture rate per year in the field was 12.76% from 1995 to 1997. Of the total 2,884 rodents and insectivores captured out- and in-doors, 527 were living rodents (including 335 A. agrarius, 119 C. triton, 6 C. barabensis, 2 C. suaveolens, 63 R. norvegicus and 2 M. musculus, and 15,467 chigger mites were collected from them. Two hundred and fifty-three of 335 A. agrarius were parasitized by chiggers, showing 75.52% (253/335) of the infestation rate and 17.53 of the chigger index; 106 C. triton were parasitized by chiggers, showing 89.08% (106/119) infestation rate and 75.93 of the chigger index. The average antibody positive rate of rodents was 14.78%. The seasonal change of the antibody positive rate was higher during December-February (the second year), and varied from 20% to 28%, but the level of antibodies remained relatively low (5.26-16.67%) during March-November. The results of serotyping with 47 antibody-positive sera were as followings: 39 sera were Gilliam types, 7 sera were Karp types, 1 serum was Kato type. Twelve strains of O. tsutsugamushi were isolated from A. agrarius (8 strains), C. triton (3 strains) and R. norvegicus (1 strain), out of the isolated 12 strains, 10 were Gilliam strains, 2 were Karp strains. A. agrarius and R. norvegicus were the main host animals in out- and indoors respectively. PMID- 15115096 TI - Early diagnosis of scrub typhus in Thailand from clinical specimens by nested polymerase chain reaction. AB - The early detection of scrub typhus in Thailand by nested polymerase chain reaction (PCR) is presented. The diagnosis of scrub typhus, from clinical samples obtained from hospitals in the northern part of Thailand, by nested PCR was compared to immunofluorescence (IF) and Weil-Felix (WF) tests. The primer pairs used for the nested PCR were designed on the basis of the nucleotide sequence of the gene that encodes the 56-kDa antigen, and RFLP analysis was used for identification. Clotted blood from 80 patients suspected of scrub typhus infection were tested. With the IF test, antibodies for Orientia tsutsugamushi were observed in 38 patients checking IgM and IgG titers. Only 21 patients showed positive seroconversion while 17 patients were negative. For the WF test, only 13 patients gave a positive seroconversion. In the early stage of infection, 19, 13 and 3 patients were detected with a sensitivity of 90.47% (19/21), 61.90% (13/21) and 14.28% (3/21) by the nested PCR, IF and WF test respectively. Two patients who were negative for seroconvesion by IF and WF were positive by nested PCR. Therefore, this suggests that nested PCR is applicable for specific rapid diagnosis at an early stage of scrub typhus in endemic regions. PMID- 15115097 TI - Characterization and utilization of monoclonal antibodies reactive to Yersinia pseudotuberculosis. AB - The 3 murine monoclonal antibodies, Yps1, Yps2 and Yps3 reactive to Y. pseudotuberculosis can be stabilized and all were found to be of IgG type. Monoclonal antibody, Yps1, recognized a glycoprotein antigen of the organism with reactivity at the 55-75 kDa region, while Yps2 and Yps3 recognized protein antigens of Y. pseudotuberculosis 65 kDa and 26-28 kDa molecular weight regions, respectively. The specificity of monoclonal antibodies was tested using dot ELISA and Western blotting with whole cell organisms or whole cell sonicated soluble antigens of different Yersinia species, Salmonella typhi, Klebsiella pnemoniae, Streptococcus abortus-equi and Escherichia coli. Monoclonal antibody, Yps1 exhibited cross-reactivity with soluble antigens and whole cell preparations of Y. pestis. Yps2 cross-reacted to soluble antigens of all the tested bacteria. Reactivity of monoclonal antibody, Yps3 was restricted to Y. pseudotuberculosis and Y. pestis with soluble antigen preparations. No reaction was observed with Yps2 and Yps3 to whole cell organism preparations from tested bacteria including Y. pseudotuberculosis. The co-agglutination reagent prepared by sensitizing staphylococcal cells with Yps1 monoclonal antibody produced a positive agglutination with all the 4 Y. pseudotuberculosis isolates and the 3 Y. pestis strains tested. Sandwich dot ELISA using monospecific antisera as a capture antibody and a monoclonal antibody, and Yps3 as a revealing antibody had a high level of specificity in detecting Y. pseudotuberculosis antigens. PMID- 15115099 TI - Prevalence of vancomycin and high level aminoglycoside resistant enterococci among high-risk patients. AB - Enterococci have been recognized as clinically important pathogens in high-risk populations of hospitalized patients. The role of enterococci in nosocomial infections is being recognized with increasing frequency. The main source of these infections is usually fecal carriage of the microorganisms. In this study, gastrointestinal colonization with vancomycin resistant enterococci (VRE) and high-level aminoglycoside resistant enterococci among 316 high-risk hospitalized patients were investigated. One hundred and ninety-eight enterococci strains were isolated from stool specimens. All strains were identified to species level and 90 of the isolates were identified as Enterococcus faecalis (45%), 85 as E. faecium (21.5%), 14 as E. avium (7%), 7 as E. raffinosus (3.5%), 1 as E. durans (0.5%) and 1 as E. hirae (0.5%). Eleven of 198 strains were found to be moderately sensitive to vancomycin (MIC: 8-16 microg/ml) by the agar dilution method according to the National Committee for Clinical Laboratory Standards (NCCLS) recommendations, and the rest of these strains were found to be sensitive (MIC < or = 4 microg/ml). Twenty-eight strains showed high-level resistance to streptomycin (2,000 microg/ml) and 26 strains were found to have high-level resistance to gentamicin (500 microg/ml). Twelve of these strains had high-level resistance to both aminoglycosides. By the disk diffusion tests, 53 of 198 strains were found to be resistant to erythromycin, 51 to penicillin, 37 to ampicillin, 18 to ciprofloxacin, 14 to norfloxacin and 3 to nitrofurantoin. No beta-lactamase production was detected in 198 studied strains. PMID- 15115098 TI - Management of infant diarrhea with high-lactose probiotic-containing formula. AB - To study the influence of high-lactose probiotic-containing formula on the course of acute diarrhea, an experiment using a randomized controlled clinical trial with patients having acute diarrhea for 3 days was conducted. One hundred patients were allocated into two groups that were comparable for age, sex, and nutritional status. The test group was administered high-lactose Bifidobacterium bifidum-containing formula, while the control group had no high-lactose probiotic until the end of the experiment. The degree of subsequent diarrhea and recovery were monitored in both groups. The results for the test and control groups were analyzed and compared using the chi-square test and Fisher exact test with a significance level (alpha) of 0.05. The study results revealed that there was no significant difference between the test and control groups (p>0.05) as well as at positive clinical test (13%) and positive floating test (65%). However, the patients receiving probiotic-containing formula had significantly less frequency of stools, when compared with the control group (p<0.05). PMID- 15115100 TI - Epidemiology and clinical picture of the Russell's viper (Daboia russelii russelii) bite in Anuradhapura, Sri Lanka: a prospective study of 336 patients. AB - Russell's viper (Daboia russelii russelii) bite is associated with a high incidence of morbidity and mortality in Sri Lanka. Hence, this study enrolled all consecutive Russell's viper bite admissions to the 'Unit A' of General Hospital, Anuradhapura, over a two year period from January 1996, to describe the epidemiology, clinical picture, treatment and outcome. There were 336 cases which showed the following results. The male: female ratio was 5:1; 75% of patients were below the age of 40 years. Biting occurred mainly in paddy fields 41%, and on footpaths 29% at dusk or dawn. Envenoming manifested in 310 (92%) of patients as follows: local swelling 92%, local necrosis 8.9%, coagulopathy 77%, neurotoxicity 78%, nephrotoxicity 18%, cardiac effects 3-12% and myotoxicity 14%. Coagulopathy appeared within 30 minutes to 12 hours after the bite and was corrected within 1 hour to 48 hours (mode 20 hours). Neurotoxicity recovered spontaneously in 1 to 5 days (mode 3 days): however, eight patients needed mechanical ventilation. Thirteen (4%) of patients were managed with peritoneal dialysis because of hyperkalemia caused by a hypercatabolic state (7) and acute renal failure (6). The mortality rate of the series was 2.6% (9 patients). Rural dry zone paddy farmers are the common victims of Russell's viper bite in Sri Lanka and its' envenoming leads to diverse clinical manifestations. Therefore, practically feasible preventive measures should be developed to minimize the incidence of bite and an evidence based management guideline should be developed for hospital practice. PMID- 15115102 TI - Tuberculous meningitis in adults: a four-year review during 1997-2000. AB - We reviewed the charts of all HIV-negative patients 15 years of age or older in whom tuberculous meningitis was diagnosed and treated without corticosteroids at Srinagarind Hospital, Khon Kaen, Thailand during the period of 1997-2000. Forty five patients were included in this study. The clinical manifestations were subacute to chronic meningitis and they presented in stages 1 and 2, except one case which was in stage 3. All patients were treated with a 6-month course of chemotherapy with good clinical outcomes. The mortality rate was 2.2% and the percent of residual neurological deficits after treatment was 6.7%. The review showed the good clinical outcomes can be had without adjunctive corticosteroid. PMID- 15115101 TI - Comparative study of the reactogenicity of a three-component acellular pertussis vaccine and whole-cell pertussis vaccine administered to healthy Singaporean infants. AB - The objective of this study was to assess and compare the reactogenicity of GlaxoSmithKline (GSK) Biologicals' diphtheria-tetanus-tricomponent acellular pertussis vaccine (DTPa) and the locally used combined diphtheria-tetanus-whole cell pertussis vaccine (DTPw) as a primary vaccination course in healthy infants at the age of 3, 4 and 5 months. A phase IV, single-blinded, randomized comparative clinical study involved one hundred and eighty healthy infants with two study groups in a 2:1 ratio to receive either DTPa or DTPw vaccine which were administered intramuscularly at the right anterior-lateral aspect of the thigh. The incidence and intensity of local solicited symptoms such as pain, redness and swelling at injection site and general solicited symptoms such as fever and fussiness were evaluated. Serious adverse events were followed for one month after each vaccination. The overall incidence of local and general symptoms was significantly higher in the group receiving locally used DTPw vaccine as compared to the group receiving GSK DTPa vaccine. Solicited local symptoms, pain (47.4% vs 15.1%), redness (95.9% vs 84.9%) and swelling (46.2% vs 18.5%), were reported more frequently in the group receiving DTPw vaccine than in the group receiving DTPa vaccine. Fever (> or = 37.5 degrees C) (52% vs 14.6%) and fussiness (60.8% vs 33.6%) were also more commonly reported in the DTPw group. There were six serious adverse events reported (4 with DTPw and 2 with DTPa). None of them related to the study vaccines, as considered by the investigators. Thus it was found that GSK Biologicals' DTPa vaccine was significantly less reactogenic as compared to the locally used DTPw vaccine manufactured by Commonwealth Serum Laboratories when administered as a 3-dose primary vaccination course to healthy infants at the age of 3, 4 and 5 months in Singapore. PMID- 15115103 TI - Telomerase activity in Malaysian patients with central nervous system tumors. AB - Telomerase, the enzyme that stabilizes telomere length is reactivated with almost all cancer types, and may be a useful diagnostic marker for malignancy. Telomerase activity has been detected in germ line cells and most cancer cells, whereas most normal somatic cells have no clearly detectable telomerase activity. In our study, we aim to detect telomerase activity in 20 human central nervous system tumors from Malaysian patients. Telomerase activity was detected based on a highly sensitive procedure consisting of a CHAPS detergent-based extraction from frozen tissues and a PCR-based telomeric repeat amplification protocol (TRAP) using a TRAPEZE Telomerase Detection Kit (Intergen, Co). Telomerase activity was considered positive when a ladder of products was observed starting at 50bp, with 6bp increments. The activity was detected in 30% of the samples analysed, included glioblastoma multiforme, meduloblastoma, paraganglioma and oligodendroglioma. The result of Fisher's exact test indicated that there was a significant association between telomerase activity status with tumor grade (p=0.003). These results suggest that telomerase activity may be an important marker for tumor malignancy. PMID- 15115104 TI - Phagocytic activities of neutrophilic leukocytes in women in various phases of menstrual cycle, and in pregnancy. AB - This study evaluated the phagocytic activities of neutrophilic leukocytes in both sexes during reproductive age and in women in different phases of the menstrual cycle and in pregnancy. In this study, 9 male and 12 female medical students, 17 to 20 years of age, 23 pregnant women and 9 women after delivery were evaluated. Heparinized blood was incubated with heat killed bacteria Staphylococcus aureus at 37 degrees C for 20 minutes. The blood films were stained with Leishman stain and the phagocytic index was found out by the number of neutrophilic leukocytes ingested with organisms in 100 cells and the total number of organisms counted therein. At the interval of 14 days, on 3 consecutive occasions, the phagocytic index was studied on the same male and female subject. The phagocytic activities of neutrophilic leukocytes were found slightly higher in females than in males; more so in pregnancy than in non-pregnant women, woman controls and after delivery. During the expected ovulation period, the phagocytic activities were observed more than during the menstrual and premenstrual phases. The phagocytic activities were found more in pregnancy and during the ovulatory period in women. PMID- 15115105 TI - Molecular markers for diagnosis of Prader-Willi syndrome in thai patients by fish. AB - Paternal microdeletion of chromosome 15 at q11-q13 has been reported in 75% of Prader-Willi syndrome (PWS) patients in western countries. Diagnosis of PWS in Thailand is mainly based on clinical observation and, in some cases, confirmed by conventional cytogenetic analysis. Loss of a tiny segment in this region (microdeletion) has made it difficult to discriminate from the normal karyotype. An attempt to solve this problem has been made by using a high resolution chromosome culture. However, this method is a tedious and time-consuming technique which is suitable for only experienced cytogeneticists. We report molecular cytogenetic analysis for PWS in Thai patients using FISH in addition to standard GTG- banding chromosome analysis. Nine Thai patients clinically diagnosed or with a suspicion of PWS were investigated. The FISH probes consist of the region-specific probes (SNRPN or D15S10 probe) and two chromosome 15 specific control probes (D15Z1 centromeric and PML chromosome 15 long arm probe). Bright field and FISH programs of an automatic karyotyper were applied to facilitate the efficiency of the chromosome analysis. We found that 2 out of 9 patients showed a deletion at 15q11-q13 region by standard GTG chromosome analysis while 4 out of 9 patients showed a delation in this region by FISH. Consistent losing of SNRPN and D15S10 signals in FISH was observed in these patients. This forty-four per cent deletion is considerably lower than those reported from western countries. We propose that DNA methylation at SNRPN promoter as well as structural abnormalities in other chromosome regions might also play a role in the etiology of this disorder in Thais, which should be investigated further. PMID- 15115106 TI - Rapid screening of PNH red cell populations using the gel test. AB - Paroxysmal nocturnal hemoglobinuria (PNH), an acquired clonal hematopoietic stem cell defect is underdiagnosed because of its atypical symptoms in some patients and because available methods, which are time consuming and complicated, are not widely used. The purpose of this study is to compare the results of the detection of PNH red cell populations using the PNH gel test and the Ham test. Fifty-eight blood samples obtained from 35 patients and 23 healthy blood donors were tested for PNH by the PNH gel test and the Ham test. It was found that 7 (20%) of the patients were positive for PNH by both tests. Twenty-three blood samples from healthy donors were all negative for PNH by both tests. The overall sensitivity and specificity of the gel test were 100%. This study showed that the PNH gel test was simple and could replace the Ham test as a screening test for PNH. This test would be especially easy to introduce in laboratories that are already using this system for blood grouping and antibody detection. PMID- 15115107 TI - Cost-effectiveness in establishing hemophilia carrier detection and prenatal diagnosis services in a developing country with limited health resources. AB - The cost-effectiveness of carrier detection and prenatal diagnosis for hemophilia at the International Hemophilia Training Center, Bangkok, Thailand was studied. From 1991 to 2002, 209 females from 124 families with hemophilia A and B were included. There were 180 hemophilia A carriers and 29 hemophilia B carriers which could be classified into 78 obligate and 131 possible carriers. The phenotypic analysis for possible carriers involved the determination of levels of factor VIII or IX clotting activity (FVIII:C, FIX:C) and the ratio of FVIII:C and von Willebrand factor antigen. The result revealed that 49 females (37.4%) were diagnosed as carriers, 65 females (49.6%) were normal and 17 females (13%) were undetermined. Additional genotypic analysis was provided to 46 families with 74 females with obligate, proven or undetermined carriers within the reproductive life. The polymorphisms associated with factor VIII and IX genes were used including Bcl I for the factor VIII gene and combined use of Mse I, Sal I, Nru I, Hha I and Dde I for the factor IX gene. The informative rate was 59.4% (44/74). Consequently, 12 prenatal diagnoses for fetus at risk were performed. Sex determination was initially determined and followed by the diagnosis of hemophilia through informative gene tracking and/or the measurement of fetal levels of FVIII:C or FIX:C. The result revealed that 3 male fetuses were affected. The total cost of carrier detection and prenatal diagnosis that the families had to pay in the government hospital was 238,600 Baht (US dollars 5,965). It was compared to the estimated cost of minimal replacement therapy using lyophilized cryoprecipitate for the survival time of 30 years in one patient with hemophilia of 1,012,500 Baht (US dollars 25,312.5). The cost of prevention was much less than the replacement therapy. In conclusion, it is cost effective to establish the service for carrier detection and prenatal diagnosis for hemophilia especially in developing countries with limited health resources. PMID- 15115108 TI - Effects of elephant garlic volatile oil (Allium ampeloprasum) and T-2 toxin on murine skin. AB - Effects of elephant garlic (Allium ampeloprasum) volatile oil (GVO) and trichothecene (T-2) toxin were studied in Swiss albino mice. The animals were 1) topically applied GVO, 2) topically applied T-2 toxin, 3) topically applied GVO followed by T-2 toxin (GVO/T-2), and 4) T-2 toxin application followed by GVO (T 2/GVO) on the right footpad. All animals were observed by Langerhans cell enumeration and pathological changes of the footpad on days 1, 3, 5 and 7. The number of Langerhans cells in the GVO treated group (1,097 +/- 33/mm2 to 1,624 +/ 19/mm2) was not significantly different when compared with the corresponding control left footpad (1,143 +/- 33/mm2 to 1,674 +/- 21/mm2). Langerhans cells density in T-2 toxin treated group (629 +/- 29/mm2to 1,090 +/- 31/mm2) was reduced by 20-35% of the opposite control footpad (962 +/- 40/mm2 to 1,392 +/- 29/mm2). Furthermore, GVO/T-2 and T-2/GVO treated mice showed a decrease in Langerhans cell number than a single T-2 toxin treated group. While Langerhans cells in T-2 toxin, GVO/T-2 and T-2/GVO groups revealed a smaller cell size with shortening dendritic processes when compare to the normal control group. Histopathological findings of the footpad skin in T-2 toxin treated group revealed epidermal desquamation and necrosis with edema and inflammatory cells infiltration. While GVO/T-2 and T-2/GVO showed a similar sequence but a lesser severe degree. These findings suggested that GVO both in pre- and posttreatment could protect T-2 toxin induced epidermal damage in a mouse footpad. PMID- 15115109 TI - Acute effects of SO2 and particles from a power plant on respiratory symptoms of children, Thailand. AB - Epidemiological studies to evaluate the acute effects of ambient SO2 on the respiratory health of children provide inconclusive results. A panel study to examine the association of short-term exposure to ambient SO2 and respiratory symptoms of 196 children for a period of 107 days was conducted in Thailand. Generalized Estimating Equations were used to examine the association of daily variation of air pollution with daily respiratory symptoms. During the study period, SO2 was not associated with respiratory symptoms in either asthmatics or non-asthmatics, whereas a 10 microg/m3 increase in PM10 was modestly associated with increases of lower respiratory symptom incidence (OR=1.03, 95%CI=0.98, 1.09) and cough (OR=1.04, 95% CI=1.00, 1.08) in asthmatics. At the low ambient air pollution concentrations observed, particulate matter rather than SO2 was associated on a microg/m3 basis with acute daily respiratory symptoms. PMID- 15115110 TI - Effect of fluoride on human dental pulp cells in vitro. AB - Human dental pulp cells were cultured in fluoride containing medium of various concentrations (0, 1, 2, 5, 10, 20, 25, 30, 40, 50, 60 and 80 ppm) in order to study the biological effect on the cells' proliferation and alkaline phosphatase (ALP) activities. It was found that fluoride at 5 ppm concentration significantly stimulated cell proliferation and ALP activity between 24 and 48 hours after exposure whereas at higher concentrations (40 - 80 ppm), fluoride significantly inhibited cell growth and ALP activity after 48 hours (Student's t test). The maximum effect was around 80 ppm. These observations suggest that fluoride, if used at a low concentration, may be a useful therapeutic agent for the treatment of pulpal disease by means of stimulating the proliferation and differentiation of dental pulp cells. At higher concentrations, it will have negative effects on this kind of cell. PMID- 15115111 TI - Advantages of trained TBA and the perception of females and their experiences with reproductive health in two districts of the Luangprabang Province, Lao PDR. AB - The study describes reproductive health in two districts of the Luangprabang Province in northern Lao PDR. The aim was to find out whether training traditional birth attendants (TBA) might have an impact on reproductive health. In June/July 2000, a total of 298 women of reproductive age, with children below two years of age, from 30 villages were interviewed by means of a closed questionnaire. In 1996/1997, a training course for TBA was conducted in one of the districts under survey. Information was obtained for demography, symptoms and risks during pregnancy and delivery, antenatal care (ANC), tetanus immunization, food taboos, place of delivery, birth attendant, practising of birth spacing and their attitude towards the services of TBA. The results obtained indirectly pointed towards a high fertility rate and a high rate of child death and abortion. An overwhelming majority of the women delivered at home, attended only by untrained individuals. During pregnancy and after delivery, the women claimed that they often suffered from edema of legs and feet, high fever and hemorrhages. Only 50% of the females in the district where TBA training were conducted, made use of the services of trained TBA. Nevertheless, females in the district with trained TBA, who made use of the TBA service in comparison with women in the same district not using the service of TBA, were 3.8 times more likely to also make use of the ANC service; 3.3 times more likely to seek immunization, and 8.6 times more likely to give colostrum to their new-borns. The educational level of the females proved to be an important factor. Literate women were more likely to practise birth spacing and have been vaccinated. Illiterate women were more likely to be at higher risk for losing a child. In the district without TBA service the loss of a child was less likely among literate than illiterate women. It is concluded that through adequately trained TBA and through their continuous support and supervision, ANC and health education can be improved. In addition to the improvement of the referral system for emergency cases and manpower development within the obstetric curative service, the training of TBA will have a positive impact on reproductive health. However, maternal health depends, to a large extent, on the educational level of the women. PMID- 15115112 TI - Determining public health priorities for an ageing population: the value of a disability survey. AB - In order to determine which diseases and health problems were most strongly associated with long-term disability among the Thai elderly and to determine their public health priority, a national cross-sectional multistage random sampling survey was conducted in 1997. Four thousand and forty-eight Thai older persons aged 60 years and over were recruited and interviewed by trained interviewers. Overall, 769 (19%) people reported having a long-term disability. Participants with long-term disability (LD) reported having between one and 21 long-term diseases or health problems. Eighteen of these problems were independently associated with LD in logistic regression analysis. Nearly half of the cases with LD (46.4%) suffered from two or more health problems. The odds of LD increased with the number of problems suffered. The problems contributing most to the population burden of disease as assessed by population attributable risk fractions were hemiparesis, arthritis, accidents (unintentional injuries), blindness and other eye diseases, kyphosis, weakness of limbs, deafness, and hypertension. This ranking of public health priority differs from conventional approaches using mortality statistics and disability adjusted life years (DALYs). In conclusion, national disability surveys provide a valuable means of assessing the population burden of disability and determining the underlying causes of disability. These methods provide a direct assessment of disability prevalence and disease priorities for rapidly ageing transitional countries where death certification may be incomplete or inaccurate. PMID- 15115113 TI - Comparison of four health systems: Cuba, China, Japan and the USA, an approach to reality. AB - The ultimate responsibility for the overall performance of a country's health system lies with government, which in turn should involve all sectors of society in its stewardship. A comparison in structure, financing and function of the health systems of Japan, the USA, Cuba and China, as well as their main health and social-demographic results, is shown here. Two clear examples of inequalities, between regions in China and between ethnic and social groups in the USA, let us see different health indicators of both countries. To achieve and maintain a healthy population with good health results does not necessarily depend on a big budget or richness in a country. Good governmental policies regarding public health and social security are crucial to achieve good quality of life equally distributed to the whole population. Some suggestions are given. PMID- 15115114 TI - Sensitive enzyme-linked immunosorbent assay with urine samples: a tool for surveillance of schistosomiasis japonica. AB - An enzyme-linked immunosorbent assay (ELISA) to detect antibodies to Schistosoma japonicum soluble egg antigens (SEA) in un-concentrated urine was developed. The urine ELISA was applied to samples collected in a schistosomiasis-endemic village in China. The levels of anti-SEA antibodies detected in urine correlated well with those obtained with paired serum samples (r = 0.694, p<0.0001). Among 129 serum ELISA positives, 112 (86.8%) were positive by urine ELISA, while all 40 serum ELISA negatives from a non-endemic area were negative. The levels of anti SEA in urine samples were stable up to 8 weeks of storage at 37 degrees C, with sodium azide as a preservative. Therefore, ELISA with urine samples can be used for the surveillance of schistosomiasis. PMID- 15115115 TI - Field assessment of recombinant Schistosoma japonicum 26 kDa glutathione S transferase in Chinese water buffaloes. AB - We have shown previously that anti-fecundity immunity can be induced experimentally against recombinant 26 kDa glutathione S-transferase (reSjc26GST) in Chinese water buffaloes (Bos buffelus), important reservoir hosts for Schistosoma japonicum in China. In the field study described here, we immunized buffaloes with reSjc26GST to induce protective immunity against S. japonicum and to evaluate its effectiveness in controlling schistosomiasis japonica. We selected two villages as test and control groups in inside-embankment areas endemic for schistosomiasis japonica. The buffaloes in the test village were vaccinated with reSjc26GST, whereas those in the control village were not. The indicators of the effect of the vaccine included the generation of specific IgG antibodies in the vaccinated buffaloes, changes in the prevalence and infection intensity in buffaloes and village children, changes in the density of infected snails, and changes in the infectivity of water bodies (assessed by sentinel mice) in transmission areas adjacent to both villages. Twenty months after vaccination, the infection rate of buffaloes in the test village was decreased by 60.4% (from an initial prevalence of 13.5% to 5.4%), and 67.9% when compared with that in the control village (initial prevalence of 16.7%). However, the infection rate in village children remained unchanged. The density of infected snails decreased by 71.4%, from 0.0049/0.11 m2 to 0.0014/0.11m2 in the high transmission area outside the embankment in the test village. There was no change in the infectivity of the water body transmission areas between the test and control villages. The levels of specific antibodies to reSjc26GST showed a continuous increase after vaccination. These results indicate that protective immunity was induced and maintained in buffaloes after vaccination with reSjc26GST. The vaccine could thus play a significant role in reducing S. japonicum transmission caused by water buffaloes in the Lake region of China. PMID- 15115116 TI - Antioxidant enzyme levels in the erythrocytes of riboflavin-deficient and Trichinella spiralis-infected rats. AB - The erythrocyte antioxidant enzyme levels of catalase, superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) of riboflavin-deficient and Trichinella spiralis-infected rats were investigated. The rats were deprived of riboflavin at the 8th week of the experiment. At that time, the erythrocyte glutathione reductase activity coefficient (EGR AC), as an indicator of riboflavin status, was > or = 1.30 in rats fed a riboflavin-deficient diet and T. spiralis-infected rats fed a riboflavin-deficient diet showed no biochemical sign of riboflavin deficiency. At the 12th week of the experiment, the levels of catalase, SOD and GSH-Px were significantly lower in the riboflavin-deficient, T. spiralis infected, and combined riboflavin-deficient and T. spiralis-infected, rats, compared to the control group. This may have been due to an increase in free oxygen radicals caused by riboflavin deficiency and parasitic infection. PMID- 15115117 TI - Trend of malaria incidence in highly endemic provinces along the Thai borders, 1991-2001. AB - The intercountry border areas of Thailand have high malaria receptivity and vulnerability that present numerous problems in the control of malaria transmission. This study focused on the 30 provinces of Thailand situated next to neighboring countries, which can be divided into 4 groups: the Thai-Myanmar border (10 provinces), the Thai-Cambodia border (6 provinces), the Thai-Lao border (10 provinces) and the Thai-Malaysia border (4 provinces). The purpose of the present study was to describe the pattern and trend of malaria incidence in the highly endemic provinces along the Thai borders for the 11 years from 1991 to 2001. Analysis of trends showed the distribution of malaria parasites to have shifted from a preponderance of Plasmodium falciparum to Plasmodium vivax along the western border with Myanmar, the northern border with Lao PDR and along the eastern border with Cambodia whereas the southern border with Malaysia the pattern changed from a preponderance of P. vivax to P. falciparum, since 1997. There was a significant difference in annual parasite incidence between borders and non-border districts, especially along the Thai-Myanmar and Thai-Cambodia borders. It is thus evident that all border districts should pay more attention to control of malaria transmission and the activities of the malaria surveillance system, and that monitoring and evaluation of the Thai Malaria Control Program needs to be performed consistently, including some areas where a few malaria cases were found as well as in malaria free areas. PMID- 15115118 TI - Distribution of two species of malaria, Plasmodium falciparum and Plasmodium vivax, on Lombok Island, Indonesia. AB - Medical and entomological surveys were conducted to determine the risk factors of Plasmodium falciparum and P. vivax infections on Lombok Island, Indonesia, to find the risk factors and the main mosquito vectors for each malaria. Multivariate longitudinal analysis demonstrated two significant risk factors for infection with P. falciparum: disappearance of P. vivax parasitemia (p<0.001) and a specific study site (p<0.001). In contrast, younger age (p=0.024) and the interpolated virtual density of An. subpictus (p=0.041) were significantly associated with increased risk of infection with P. vivax. Thus, it seems that the distribution of P. vivax was determined largely by the presence of An. subpictus, whilst that of P. falciparum was influenced by antagonism with P. vivax. This result shows the importance of following-up treated P. vivax patients to identify recrudescence of P. falciparum in this area. PMID- 15115119 TI - Malaria, leprosy and dapsone. AB - Although the preventive action of dapsone against P. falciparum malaria was known for many years, there was no report about the incidence of P. falciparum malaria in leprosy patients treated with dapsone, especially from areas of Southeast Asia where both leprosy and malaria are endemic. Therefore, two clinic-based malaria surveys were undertaken at a gap of 12 years, comprising 506 lepromatous leprosy patients and 499 febrile nonleprosy control subjects. Both the surveys showed that the lepromatous patients treated with MDT had only P. vivax malaria (incidence comparable to the febrile nonleprosy controls) with complete freedom from P. falciparum. On the contrary, control sujects not taking any-leprosy drugs and staying with the leprosy patients at the same beggars' home, had both P. vivax and P. falciparum malaria. It is postulated that dapsone provided protection against P. falciparum among leprosy patients. PMID- 15115120 TI - Some entomological observations on temporal and spatial distribution of malaria vectors in three villages in northwestern Thailand using a geographic information system. AB - This spatial and temporal heterogeneity in the distribution of Anopheles mosquitos were studied during August 2001 to December 2002 in three villages Ban Khun Huay, Ban Pa Dae, and Ban Tham Seau, in northwestern Thailand in Mae Sot district, Tak Province. The three Karen villages are located about 20 km east of the city of Mae Sot near the Myanmar border. Twenty-one species were collected on human collections during 68 nights of 17 months. Anopheles minimus comprised of 86% of the specimens biting man. An. minimus was implicated as a vector based on the detection of sporozoite infections using enzyme-linked immunosorbent assays for Plasmodium falciparum and P. vivax. Seasonal comparison of vectorial capacity and entomological inoculation rate was calculated. An. dirus was rarely encountered and probably played little part in transmission in these three villages during the period of study. Information is provided on nightly biting activity, parity rate, infectivity, and adult and larval bionomics. Spatial and temporal comparisons among the collections were displayed on different satellite images including the Normalized Difference Vegetation Index data from on the National Oceanographic and Atmospheric Administration satellites (NOAA/NDVI), the LANDSAT satellite Thematic Mapper (spatial resolution 30x30 m) and the IKONOS satellite (spatial resolution 1x1 m) in a Geographical Information System (GIS). PMID- 15115121 TI - Use of GIS-based spatial modeling approach to characterize the spatial patterns of malaria mosquito vector breeding habitats in northwestern Thailand. AB - We sampled 291 bodies of water for Anopheles larvae around three malaria-endemic villages of Ban Khun Huay, Ban Pa Dae, and Ban Tham Seau, Mae Sot district, Tak Province, Thailand during August 2001-December 2002 and collected 4,387 larvae from 12 categories of breeding habitat types. We modeled surface slope and wetness indices to identify the extent and spatial pattern of potential mosquito breeding habitats by digitizing base topographical maps of the study site and overlaying them with coordinates for each larval habitat. Topographical contours and streamlines were incorporated into the Geographical Information System (GIS). We used Global Positioning System (GPS) instruments to locate accurately each field observed breeding habitat, and produced a 30-m spatial resolution Digital Elevation Model (DEM). The slope (of less than 12 degrees) and wetness (more than 8 units) derived from spatial modeling were positively associated with the abundance of major malaria vectors An. dirus, An. maculatus, An. minimus, and An. sawadwongporni. These associations permit real-time monitoring and possibly forecasting of the distributions of these four species, enabling public health agencies to institute control measures before the mosquitos emerge as adults and transmit disease. PMID- 15115122 TI - Larval habitats and distribution patterns of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse), in Thailand. AB - This study was conducted to survey larval breeding habitats and to obtain larval abundance during the dry period covering all 5 geographical zones of Thailand. Our results indicated Aedes aegypri is prevalent all over the country, whereas Aedes albopictus is more restricted to the remote area of the south. Water storage containers, especially water jars, served as a main larval breeding habitats of Ae. aegypti, whereas broken cans and plastic containers are considered primary breeding sites for Ae. alpopictus during the dry period. In addition, Aedes larval indices, container index (CI), house index (HI), and Breteau index (BI) were measured. CI and HI values from the central part were significantly higher than those from other areas (p<0.01). BI values of all collection sites were greater than 50 (a maximum BI value accepted by the Ministry of Public Health, Thailand). In brief, Ae. aegypti and Ae. albopictus populations heavily infested many towns and residential areas of the country. Drought could not limit the density of Aedes mosquitos in Thailand. Systematic vector control and vector surveillance programs by public health organizations, if practical, should be continuously conducted to reduce or prevent dengue risk. PMID- 15115123 TI - Monitoring the therapeutic efficacy of antimalarials against uncomplicated falciparum malaria in Thailand. AB - Increasing antimalarial drug-resistance is an important problem in Thailand. The results of monitoring the antimalarial efficacy are used in decision-making about using antimalarials to treat uncomplicated falciparum malaria in Thailand. In 2002, 552 patients with uncomplicated malaria were treated according to the Thai National Drug Policy, with mefloquine 25 mg/kg plus artesunate 12 mg/kg and primaquine 30 mg in divided doses for 2 days in high-mefloquine-resistant areas; mefloquine 15 mg/kg plus primaquine 30 mg in non- or low-mefloquine-resistant areas; mefloquine 15 mg/kg plus artesunate 12 mg/kg and primaquine 30 mg in divided doses for 2 days or Coartem (6-dose regimen for adult contains 480 mg artemether and 2880 mg lumefantrine) plus primaquine 30 mg given over 3 days in moderate-mefloquine-resistant areas. The study shows that mefloquine, artesunate plus mefloquine, and artemether plus lumefantrine are effective in the treatment of uncomplicated malaria in most areas of Thailand except for Ranong and Kanchanaburi, where the first-line treatment regimen should be revised. PMID- 15115124 TI - Six-years monitoring the efficacy of the combination of artesunate and mefloquine for the treatment of uncomplicated falciparum malaria. AB - Plasmodium falciparum in Thailand is multi-drug resistant. In a previous study it was shown that artesunate and mefloquine were effective, as follow up, we monitored the efficacy of this regimen for six years. During 1997-2002, 516 adult male volunteer patients in Chanthaburi Province were enrolled (50 patients in the first year, 400 patients in 1998-2001 and 66 patients in 2002). The symptom complex and parasite count (thick blood film) were monitored on days 0, 1, 2, 7, 14, 21, 28, 35 and 42. The dosages used were artesunate (ATS) 150 mg and mefloquine (M) 750 mg at hour 0 and ATS 100 mg and M 500 mg at hour 24. Their ages ranged from 30-35 years and their mean body weights were 54-56 kg. The presenting symptoms were fever 100%, headache 97-100%, anorexia 78-90%, and nausea 28-40%. The geometric mean of parasitemia ranged from 7,357-12,750/mm3. Defervescence in one day was found in 42-76% of patients and 85-100% in 2 days. The sensitivity (S) ranged from 87-94% and RI resistance (recrudescence) ranged from 6-13%. Forty patients demonstrated RI type of response, 37 were cured after being retreated with the same dosage and another 3 patients were cured after the third course of treatment. The aggravated adverse effects included vomiting (8 20%), anorexia (1-41%) and diarrhea (0-16%). These side effects were mild and transient. The efficacy of the artesunate and mefloquine combination for the treatment of uncomplicated falciparum malaria was high. The RI type of response was possibly due to re-infection or multiple broods and not to drug resistance. The adverse effects of anorexia, nausea, vomiting and diarrhea were mild and transient for mefloquine. The combination can be used as stand by treatment in areas of multi-drug resistant falciparum malaria. PMID- 15115125 TI - In vitro susceptibility of Plasmodium falciparum isolates to chloroquine and mefloquine in southeastern Mindanao Island, the Philippines. AB - Although the presence of multi-drug-resistant falciparum malaria has been reported in the Philippines, the distribution of drug-resistant malaria parasites has not yet been determined in Mindanao Island. In vitro susceptibility of P. falciparum to both chloroquine and mefloquine was assessed to forecast the spread of drug-resistant parasites in various foci in southeastern Mindanao Island. Of the 33 isolates of P. falciparum successfully tested, 10 (30%) were susceptible, 12 (36%) showed decreased susceptibility (80 nM < or = IC50 < 114 nM), and 11 (33%) were resistant (IC50 > or = 114 nM) to chloroquine. Ten (91%) of the resistant isolates and 9 (75%) of those with decreased susceptibility were from northern and northwestern Davao del Norte Province. Chloroquine-susceptible isolates were found among patients in the eastern parts of Davao del Norte and Davao Oriental provinces. Seven isolates from several foci in the study area were all mefloquine- susceptible (IC50 < 10 nM). This is the first report indicating the potential emergence of chloroquine-resistant P. falciparum on Mindanao Island, which is presently regarded as a drug-susceptible area. PMID- 15115126 TI - Lack of in vitro effect of ivermectin on Plasmodium falciparum. AB - The in vitro activity of ivermectin was assessed against the K1 isolate of Plasmodium falciparum. The mean IC50 and IC90 of ivermectin were 8.0 and 35.0 microg/ml, respectively. These results indicate that ivermectin has a very low activity against P. falciparum in vitro. PMID- 15115127 TI - NS2a-NS2b sequence analysis of DEN-4 virus strains. AB - To study the genetic variability of DEN-4 Chinese isolates, and to trace the origin of DV4 Chinese isolates, we cloned and sequenced the NS2a-NS2b junction of 5 isolates and prototype DV4 (H-241). Our results show that isolates from the 1990 Guangdong epidemic, which were isolated in the early, middle, and late periods of the epidemic, share the same sequence in the NS2a-NS2b junction. The sequence similarity between isolates from the Guangdong epidemic in 1990 and DV4 H-241 is 96%; these isolates can be grouped into genotype I. The sequence similarity between the isolate from the Guangdong epidemic in 1987 and Dominica strain 814669 is 96%; this isolate can be grouped into genotype II. For the first time, our results show that there are also 2 DV4 genotypes in the Guangdong area of southern China, and these isolates perhaps were introduced from other epidemic areas outside of China. PMID- 15115128 TI - A glance at the von Willebrand factor in dengue virus infection. AB - To identify the level of von Willebrand factor (vWF) in dengue infection, especially severe DHF, and correlate the increase in vWF with thrombocytopenia, children admitted with dengue fever/DHF were examined for hemoglobin, hematocrit, platelet, and vWF for three consecutive days. Anti-dengue IgM and IgG were determined. Correlations between vWF and thrombocytopenia were analyzed using multivariate analysis. Forty-one patients were eligible for the study; of whom almost three fourths had a secondary infection, as proved serologically. At the beginning of the study, a high level of vWF along with a low platelet count were seen, which seemed to fit the hypothesis that an increase in vWF in the serum will be followed by a decrease in platelets, as a result of the platelet aggregation process in the peripheral blood vessels, predisposed by the immune complex events in DHF. Observations for three consecutive days revealed significant changes of vWF levels (p = 0.000) as well as platelet counts (p = 0.002). However in the context of dengue infections, these changes did not correlate well (p = 0.988). Could there have been a significant correlation if cases were followed for a longer period of time? Being a part of a more comprehensive study, it appeared that in patients with dengue infections, vWF and platelets were not the only factors involved in bleeding, indicating that activation of endothelium is one factor in a multifactorial process. PMID- 15115130 TI - Sero-epidemiological study of hepatitis A virus infection among hill-tribe youth and household environmental sanitation, a hill-tribe community in northern Thailand. AB - A cross-sectional analytic study of 190 hill-tribe youth in a community in the north of Thailand was conducted to investigate the sero-prevalence of HAV and factors related to positive anti-HAV antibody. The studied youth, whose ages ranged from 15 to 24 years, were interviewed about socio-economic status and personal hygiene. Blood specimens were collected to detect anti-HAV by ELISA commercial kit. Household environmental sanitation conditions were observed and drinking water samples were screened for bacterial contamination using SI2 medium. Following the anti-HAV assay, the studied youth were divided into two groups: anti-HAV positive, and anti-HAV negative. The studied variables of the two groups were analyzed by chi2 test to find factors related to anti-HAV positivity. The results revealed that 87% of the studied youth were positive for anti-HAV. There was no statistically significant difference between age group/gender and anti-HAV positivity, p = 0.46 and 0.16, respectively. Approximately 35.79 to 45.79% washed their hands with soap before preparing food, before eating and after using the latrine. About 88% did not improve the potability of their drinking water. The results of screening for bacterial contamination in drinking water samples found that 73.53% were contaminated with coliform bacteria. Factors related to positive anti-HAV antibody included monthly income, number of household members, use of latrine, hand-washing with soap after using latrine, household refuse management and control of insects and rodents; p = 0.04, 0.007, 0.013, 0.008, <0.001 and <0.001, respectively. The findings suggested that appropriate household environmental management should be improved in this community to reduce HAV transmission. PMID- 15115129 TI - Seroepidemiological survey among schoolchildren during the 2000-2001 dengue outbreak of Ratchaburi Province, Thailand. AB - From August 2000 to 2001, a dengue outbreak occurred in Mueang district, Ratchaburi Province, Thailand. About 800 cases of dengue infection were reported, and among them, 49.5% were clinically diagnosed as dengue hemorrhagic fever according to the WHO criteria. During the outbreak, the incidence rate of dengue infection in Hin Gong subdistrict was 2.9 per 1,000 population. A seroepidemiological survey was conducted among primary schoolchildren from July 2000 to June 2001, to monitor dengue transmission. In a baseline survey, 283 children were surveyed for dengue antibody and 71% were IgG seropositive. In June 2001, the rate of dengue infection showed an increase of 8.8% with 8.0% among immune children and 10.3% among naive schoolchildren. Among 283 schoolchildren, 90 were followed up 3 times, in September and December 2000, and June 2001. An increase in the rate of seroconversion was observed in the period September to December 2000, while the peak dengue outbreaks in the dry season occurred in February 2001. Serosurveys among schoolchildren appear to be early warning system, and can be advantageous in early dengue control actions, in order to break the chain of transmission before an impending epidemic. PMID- 15115131 TI - Pneumocystis carinii infection among human immunodeficiency virus (HIV) infected Myanmar patients. AB - A total of 60 HIV infected patients complaining of dry cough for at least two weeks and attending the Out-patient Department of the Specialist Hospital, Waibargi, were screened for Pneumocystis carinii. Induced sputum samples were examined with Giemsa and Gomori silver methenamine stains. P. carinii were detected in 18 patients (30%) with silver stain and 13 patients (21.7%) with Giemsa stain. The sensitivity and specificity of the Giemsa stain were 72.2% and 95.2%, respectively. The range of CD4 counts in P. carinii-positive patients was found to be 0-562/microl, and the mean CD4 count was 132.3/microl. Out of 18 P. carinii-positive cases, CD4 counts of 15 cases (83.3%) were <200/microl and those of 3 cases were >200/microl. Clinically, P. carinii-positive cases were associated with fever in 55.5%, with tightness of the chest in 38.9%, and with cyanosis and tightness of the chest in 11.1%. Co-infection with tuberculosis was found in 16.7%. Anti-pneumocystic prophylaxis is recommended for those patients with a CD4 count <200/microl. Giemsa staining could be used as an alternative diagnostic method for detecting P. carinii. This study documented the existing prevalence of P. carinii among HIV-infected Myanmar patients. PMID- 15115132 TI - High frequency of HIV-1 and hepatitis C co-infection among young Thai men: evidence for a changing pattern of HIV transmission in Thailand. AB - To assess whether patterns of HIV transmission have changed in Thailand, we tested for antibody to hepatitis C virus (HCV) as a marker for parenterally acquired infection among HIV-infected and uninfected young Thai men. Antibody to HCV was present in 49.5% of HIV-infected men and 2.2% among uninfected men. These data suggest that a significant number of HIV infections among young men in Thailand may be associated with injection drug use. PMID- 15115133 TI - Rectal prolapse associated with cytomegalovirus pseudomembranous colitis in a child infected by human immunodeficiency virus. AB - We report a newly recognized presentation of cytomegalovirus (CMV) enterocolitis in a 4-year-old girl with newly diagnosed HIV disease who presented with rectal prolapse. Gross findings showed multiple whitish punctate lesions. An endoscopic examination revealed multiple shallow ulcers and pseudomembranes along the colon. A biopsy from colonic tissues demonstrated CMV-like inclusion bodies. A direct immunofluorescence assay using specific CMV monoclonal antibody was positive for CMV-infected cells in specimens from the rectal smear. PMID- 15115134 TI - Prevalence of parvovirus B19 infection in Thai young adults. AB - The prevalence of antibody to human parvovirus B19 in 128 Thai healthy young adults was measured. Antibodies of the immunoglobulin G (IgG) class were investigated in serum samples of 51 males and 77 females aged 18-24 years (mean 19.83; SD 1.07) by a commercial enzyme-linked immunosorbent assay (ELISA) using high specific recombinant parvovirus B 19 antigen. Only 14 out of 128 (10.94%) sera were found positive, including 6 males and 8 females. No sex preponderance was observed. The amount of antibody calculated as antibody index was not statistically significant difference between genders. PMID- 15115135 TI - Immunogenicity and safety of a varicella vaccine, Okavax, and a trivalent measles, mumps and rubella vaccine, MMR-II, administered concomitantly in healthy Filipino children aged 12-24 months. AB - This trial was conducted to assess the immunogenicity and safety of the varicella vaccine, Okavax, when administered concomitantly with the measles, mumps and rubella vaccine, MMR-II, to children aged 12-24 months. A total of 299 children were randomized into three groups, those receiving Okavax only, MMR-II only, or both vaccines concomitantly. Antibody titers were determined by ELISA in blood samples taken immediately before, and 6 weeks after, vaccination. Parents recorded local and systemic reactions. Okavax elicited similar varicella seroconversion rates (> or = 93.9%) and high GMTs when given alone or with MMR-II (99.6 and 95.7 mIU/ml, respectively). The seroconversion rates (measles and rubella 100%, mumps > or = 75.0%) and high GMTs elicited by MMR-II were not affected by concomitant administration of Okavax. The incidence of adverse events was similar whether MMR-II and Okavax were administered concomitantly or separately, and the majority of local reactions were mild and transient, with fever the most frequent systemic event in all groups. In conclusion, these results show that the immune response and the reactogenicity profile of Okavax and MMR-II were similar when given together or alone. Concomitant administration of these vaccines can therefore be recommended for children in their second year of life. PMID- 15115136 TI - Progress and challenges toward poliomyelitis eradication in Indonesia. AB - Poliomyelitis is one of few diseases that can be eradicated. The virus cannot survive outside the body and effective vaccine is available to protect children and stop transmission. Today, there are 3 million children each year saved by the oral polio vaccine (OPV) and globally the reported cases have declined from 50,000 to 7,000 in 1999. At present, 20 countries may remain at risk of continued transmission, mostly in Africa and Asia. In the region of SEARO, wild poliovirus is still transmitted in India, Nepal and Bangladesh. Efforts to eradicate polio had been made in Indonesia, through a four-pronged strategy; routine immunization, National Surveillance Days (NIDs), surveillance of Acute Onset of Flaccid Paralysis (AFP) cases, and supplementary immunizations. No polio cases have been detected in Indonesia since 1995, but some problem will remain until the whole SEARO region is certified polio-free. Filling the immunization gap and revitalizing the AFP surveillance program are among the highest priority activities. PMID- 15115137 TI - Vomiting, abdominal distention and early feeding of banana (Musa paradisiaca) in neonates. AB - The objective of this cohort study was to assess the relationship between banana given as early solid food with the symptoms of intestinal obstruction (SIO) among neonates, in a rural community in West Lombok District, West Nusa Tenggara Province, Indonesia. Mothers having newborn infants were interviewed and 3,420 neonates were followed for 28 days. Compared with infants who were not given solid food, the relative risk (RR) for infants given food other than banana as early solid food was 1.87, 95% CI 0.48-8.24, p=0.4, while for infants given banana only as early solid food the RR was 9.15, 95% CI 1.96-42.58, p 0.0005. After adjustment for birthweight, colostrum, and breastfeeding, the odds ratio for infants given banana and the appearance of SIO was 2.99, 95% CI 2.65-5.14; p=0.0012. These data indicate that banana given as early solid food is an important risk factor for the appearance of SIO in neonates. PMID- 15115138 TI - Airlangga scoring system for prediction of dehydration in diarrheal patients. AB - To investigate the predictive factors for dehydration in acute diarrheal patients, this case control study was conducted using the observational analytic method. Acute diarrheal patients who were admitted to the Hospital and Outpatient Pediatric Clinic, Dr Soetomo Hospital, were included in this study. By discriminant analysis, three significant variables were determined to differentiate dehydration risk in acute diarrheal patients: frequency of stool, amount of feces in the stool, and severity of vomiting (power test: 70.0%). Significant differences were found between the groups with and without dehydration for stool frequency each day (p<0.05), amount of stool per day (p<0.05), and severity of vomiting (p<0.05). Frequency of stool, amount of stool, and severity of vomiting are predictive factors for dehydration in acute diarrhea. PMID- 15115140 TI - Stability of meropenem in normal saline solution after storage at room temperature. AB - The bactericidal activity of meropenem is determined by the time that concentrations in tissue and serum are above the MIC for the pathogens during the dosing interval. Thus, the most effective mode of administering of meropenem is continuous infusion. However, the stability of meropenem reconstituted in solution is influenced by the storage temperature. Until now we have had no data to evaluate the stability of this drug during continuous infusion in a tropical country. The objective of this study was to provide such data. Meropenem 0.5 g and 100 ml normal saline solution were mixed together and stored at room temperature for 8 hours. Half of the solution was stored in a room with air conditioning at 20 degrees C and the other half of the solution was stored in a room without air conditioning at 32 degrees-37 degrees C. The concentrations of meropenem in the solution were measured at 0, 1, 2, 3, 4, 6 and 8 hours after the drug was reconstituted. Twelve lots of (0.5 g meropenem in normal saline) solution were evaluated in each temperature condition. The mean meropenem concentrations reconstituted in normal saline solution decreased 1.66%, 3.31% and 5.80% after 2, 4 and 8 hours storage at 20 degrees C, respectively. Drug concentrations decreased 3.14%, 5.86% and 11.85% after 2, 4 and 8 hours storage at 32 degrees-37 degrees C, respectively. Therefore, we conclude that this agent should not be administered by 8-hour continuous infusion at room temperature in a tropical country. PMID- 15115139 TI - Antibiotic resistance, plasmid profile and RAPD-PCR analysis of enteropathogenic Escherichia coli (EPEC) clinical isolates. AB - Enteropathogenic Escherichia coli (EPEC) is a leading cause of diarrhea among infants in developing countries. A total of 38 EPEC isolates, obtained from diarrhea patients of Hospital Miri, Sarawak, were investigated through plasmid profile, antibiotic resistance and randomly amplified polymorphic DNA (RAPD) analysis. From the 8 types of antibiotics used, all isolates were 100% resistant to furoxime, cephalothin and sulphamethoxazole and showed high multiple antibiotic resistant (MAR) indexes, ranging from 0.5 to 1.0. In plasmid profiling, 22 isolates (58%) showed the presence of one or more plasmids in the range 1.0 to 30.9 mDa. The dendrogram obtained from the results of the RAPD-PCR discriminated the isolates into 30 single isolates and 3 clusters at the level of 40% similarity. The EPEC isolates were highly diverse, as shown by their differing plasmid profiles, antibiotic resistance patterns and RAPD profiles. PMID- 15115141 TI - Comparison of the effectiveness of baits used in traps for adult fly collection. AB - The effectiveness of 6 different baits used in fly traps for adult fly collection was compared in October-November, 2000, near the cafeteria of Maharaj Nakhon Chiang Mai Hospital, Chiang Mai Province, northern Thailand. The baits used were fresh viscera (pork, beef and fish) and fresh meat (pork, beef and fish), with 500 g of each kind being used in each fly trap for each experiment. All were replicated four times. Pork viscera yielded the highest effectiveness (35.2%), followed by fish meat (30%). Four other kinds of bait showed relatively low efficiency, ie fish viscera (12.0%), beef viscera (9.0%), pork meat (8.4%) and beef meat (5.4%). The calliphorid Chrysomya megacephala was the most abundant fly species collected, while sarcophagids and muscid flies were found in much smaller numbers. The number of females was higher than males in all collections. Although all baits used were parts of animal carcasses that were very attractive for adult flies to feed upon and oviposit, the result of this study showed the differences in their effectiveness for fly collection, particularly during the late rainy season to early winter in Chiang Mai. Pork viscera showed the highest efficacy for metallic fly collection, but not for all species. Further study of suitable baits for the medically important fly species that exist in urban areas should, therefore, be performed. PMID- 15115142 TI - Breakthrough neurological manifestation during appropriate antituberculous therapy of miliary tuberculosis. AB - We report a 20-month-old girl with miliary pulmonary tuberculosis and normal neurological findings. While on treatment with isoniazid, rifampicin, pyrazinamide, and ethambutol for 1 month, she developed weakness of the lower extremities without meningism or altered consciousness. A computerized tomogram revealed tuberculomas and basal arachnoiditis. The cerebrospinal fluid findings were compatible with tuberculous meningitis. She responded well to systemic corticosteroids. PMID- 15115143 TI - Cerebral infarction and cerebral salt wasting syndrome in a patient with tuberculous meningoencephalitis. AB - A 38-year old female with underlying systemic lupus erythematosus was admitted with tuberculous meningoencephalitis. After an initial good response to anti tuberculous treatment, she developed cerebral infarction and profound hyponatremia. This was due to cerebral salt wasting syndrome, which has only previously been described in 2 cases. The difficulties in diagnosis and management of this case are discussed. PMID- 15115144 TI - A simple method for extraction and purification of genomic DNA from dried blood spots on filter paper. AB - We have developed an efficient and simple method for extracting and purifying genomic DNA from dried blood stored on filter paper. The quality of the genomic DNA extracted is tested by PCR amplification of a 255-bp fragment of the PAX8 gene sequence and the PCR products are determined for further genetic studies by single strand conformation polymorphism (SSCP) analysis. Larger DNA sequences of the 674-bp of the PAX8 gene and the 1,039-bp of the human beta-globin gene, a housekeeping gene, have also been amplified from the extracted DNA, thus indicating the high quality of the genomic DNA extracted by the developed method for subsequent genetic studies of any gene of interest. The method developed can also be used for the purification of genomic DNA from dried blood specimens stored under different conditions. Moreover, the genomic DNA products can be stored for long-term use due to the highly purified procedure. Therefore, the method is efficient and appropriate for the extraction and purification of genomic DNA from dried blood specimens, which has become an increasingly important tool for genetic and epidemiological studies. PMID- 15115146 TI - Human anion exchanger1 mutations and distal renal tubular acidosis. AB - The human anion exchanger 1 (AE1 or SLC4A1) gene encodes anion exchanger 1 (or band 3) protein in erythrocytes and in alpha-intercalated cells of the kidney. Thus, AE1 mutations show pleiotrophic effects resulting in two distinct and seemingly unrelated defects, an erythrocyte abnormality and distal renal tubular acidosis (dRTA). Southeast Asian ovalocytosis (SAO), a well-known red blood cell (RBC) defect, which is widespread in Southeast Asian regions, is caused by AE1 mutation due to a deletion of 27 base pairs in codons 400-408 (delta400-408) leading to an in-frame 9 amino-acid loss in the protein. Co-existence of SAO and dRTA is usually not seen in the same individual. However, the two conditions can co-exist as the result of compound heterozygosities between delta400-408 and other mutations. The reported genotypes include delta400-408/G701D, delta400 408/R602H, delta400-408/deltaV850, and delta400-408/A858D. The presence of dRTA, with or without RBC abnormalities, may occur from homozygous or compound heterozygous conditions of recessive AE1 mutations (eg G701D/G701D, V488M/V488M, deltaV850/deltaV850, deltaV850/A858D, G701D/S773P) or heterozygous dominant AE1 mutations (eg R598H, R589C, R589S, S613F, R901X). Codon 589 of this gene seems to be a 'mutational hot-spot' since repeated mutations at this codon occurring in different ethnic groups and at least two de novo (R589H and R589C) mutations have been observed. Therefore, AE1 mutations can result in both recessive and dominant dRTA, possibly depending on the position of the amino acid change in the protein. As several mutant AE1 proteins still maintain a significant anion transport function but are defective in targeting to the cell surface, impaired intracellular trafficking of the mutant AE1 is an important molecular mechanism involved in the pathogenesis of dRTA associated with AE1 mutations. PMID- 15115145 TI - Comparative study of anti-double stranded DNA detection by ELISA and Crithidia luciliae immunofluorescence. AB - Two commercial enzyme-linked immunosorbent assays (ELISA) and two commercial Crithidia luciliae immunofluorescence tests (CLIF) were reevaluated as to the efficiency and degree of correlation of anti-double stranded DNA (anti-dsDNA) detection in systemic lupus erythematosus (SLE). The two ELISAs exhibited an overall agreement of 95% and significantly correlated with each other (r=0.91, p<0.001). They were comparable in sensitivity (64%, 61%) and had the same specificity (95%, 95%). The sensitivity of the two CLIFs was 39% and 29% with corresponding specificities of 100% and 97%, and an overall agreement with each other of 94%. The two ELISAs had comparable specificity to the CLIFs with good agreement (84%, 79%) while they had a much greater sensitivity than the CLIFs. These findings suggest that ELISA is a useful laboratory test for anti-dsDNA detection of SLE due to its simplicity, quantitative results, sensitivity, specificity and cost, as compared to CLIFs. PMID- 15115147 TI - Amyotrophic lateral sclerosis and diabetes on Guam: changing patterns of chronic disease in an island community. AB - Amyotrophic lateral sclerosis (ALS) occurred on the island of Guam with unusually high incidence rates for many years but began to disappear with the island's westernization after WW II. The authors document these changes and suggest that they support a hypothesis that cultural changes could be responsible for both the virtual disappearance of this chronic degenerative neurologic disease as well as a concurrent surge in the prevalence of diabetes. PMID- 15115148 TI - Health seeking behavior among insured persons under the Social Security Act, 1990. AB - Since having health insurance cannot guarantee access to care among the insured persons, their actual health seeking behavior should be evidence reflecting true access. Therefore, the study aimed to present the patterns of health seeking behavior among the insured persons who actually were able to get free services from their registered hospitals under the Social Security Scheme. Purposive sampling was done of 1,003 insured persons who were willing to participate in the study from small, medium and large establishments in the Huai Khwang district in Bangkok. A health diary was employed as one of the data collecting tools with a follow-up period of six months. The average illness rate found was 6.44 episodes/person/year. The characteristics of illnesses reported were described in terms of symptom groups, perceived severity, duration, work or non-work related cause. No treatment or self care, seeking help from non-registered health facilities and seeking help from registered hospitals and clinics were the patterns of health seeking behaviors found in the study. The patterns of health seeking behaviors among the participants varied depending on the stage of treatment, perceived severity of illness and types of additional health benefits. Seeking care from registered hospitals and clinics was found among the illnesses with a higher level of perceived severity, among the participants with chronic diseases, and among the illnesses that were treated with higher stages. Therefore, health insurance might not be able to guarantee true access to needed care for people unless the comprehensive health care provider networks are designed to cover more types of services, be more convenient and have more accessible health care providers. PMID- 15115149 TI - Health-related quality of life after hip fracture in the elderly community dwelling. AB - Hip fracture occurs commonly in older individuals and can have a considerable impact on the functional independence and quality of life for older patients living in the community. In a population-based case-control study six months after hip fracture, we investigated the association between functionality and quality of life. The Functional Independence Measure (FIM), the Frenchay Activities of Daily Living Index (FAI), and the Berg Balance Scale (BBS) were used to measure physical function, and quality of life was measured by completing a Short Form-36 (SF-36). With age and gender match, the hip fracture group scores were significantly lower (p < or = 0.05) than the control group in all measurements of physical function (FIM 95.54 vs 103.5; FAI 23.68 vs 30.76; BBS 46.21 vs 54.25). The quality of life was assessed by SF-36, which has eight domains: physical function, physical role, bodily pain, mental health, emotional role, social function, general health and vitality. All eight domains were significantly lower in the hip fracture group compared with the controls (p<0.05). The reduction in function was reflected in a reduction in the quality of life. Thus, clinically reported hip fracture impairs both the functionality and quality of life of these subjects. The adverse impact of hip fracture on quality of life and functionality needs to be recognized by health personnel in the community, so that adequate health resources can be devoted to preventing and treating this debilitating condition. PMID- 15115150 TI - Platelet fatty acids in coronary heart disease, dyslipidemia, hypertension and healthy controls. AB - A cross-sectional study was performed to investigate 250 volunteers from Pramongkutklao Hospital, Samphanthawong district, Wat Chaiyapreukmala and Wat Pradoo in Taling Chan district. They were divided into groups of 35 apparently healthy males, 16 males with coronary heart disease, 37 males with dyslipidemia and 9 males with hypertension with age ranges of 24-62, 56-69, 25-69 and 26-75 years, respectively. The female groups were composed of 55 apparently healthy females, 10 females with coronary heart disease, 73 females with dyslipidemia and 15 females with hypertension with age ranges of 27-65, 33-67, 22-73 and 38-70 years, respectively. Platelet fatty acids levels were found to have no significant difference between the different male groups. In the female groups, the alpha-linolenic acid (ALA) level in hypertension was significantly higher than in coronary heart disease (CHD) (p<0.05), whereas the arachidonic acid (AA) level in hypertension was significantly higher than in the apparently healthy females (p<0.05). No correlation was found between platelet fatty acids and age or anthropometric parameters, which indicate that platelet fatty acids may not depend on either age or anthropometric parameters. Positive correlations were shown between ALA and eicosapentaenoic acid (EPA), AA and docosahexaenoic acid (DHA), ALA and the diastolic blood pressure, DHA and total cholesterol (TC), and between low density lipoprotein choleoterol (LDL-C) and plasma glucose. Negative correlations were shown between LA and EPA, AA and EPA, EPA and DHA, EPA and the systolic blood pressure, and AA and the diastolic blood pressure. PMID- 15115151 TI - The validity of peer responses as a tool for screening at-risk students: a preliminary analysis. AB - Students are becoming the majority of new amphetamine users in Thailand. This study compared urinalysis results with peer responses to individual characteristics related to substance use with the aim of identifying "at-risk" students. A randomly selected group of students from a public high school in northern Thailand was asked to fill out the names of classmates they viewed as having any of forty-three risk behaviors set out in a questionnaire. A total of 564 students were included, from whom urine specimens were collected on the first two days following the school break. An immunoassay test was used to screen the specimens and positive results were confirmed using thin-layer chromatography. About 4% of urinalysis results were methamphetamine-positive. Using urine test results as the standard, the sensitivity of peer responses to alienated behavior was 81.8%, while frequent class/school absenteeism and low concentration levels were somewhat lower, at 77% and 68%, respectively. Delinquency showed the least sensitivity at 50%. The McNemar chi2 test showed significant differences between urine test results and each peer response subscale (p < 0.001). This preliminary analysis has shown that peer responses with regard to substance-related behavior compare well with urine test results. PMID- 15115152 TI - Report on blood cholinesterase among vegetable growers. AB - We report our study on serum cholinesterase in vegetable growers, a risk occupation in a rural area of Thailand. In this study, 70 subjects (35 vegetable growers and 35 controls) were studied. The mean blood cholinesterase level in vegetable growers (17.7 +/- 7.0 U/ml) was significantly lower than that of the control group (24.7 +/- 12.4 U/ml) (p=0.01). PMID- 15115153 TI - How we do it: CT of the spleen. PMID- 15115154 TI - Case of the month. Abdominal pregnancy. PMID- 15115155 TI - Computed tomographic angiography of endovascular abdominal aortic stent-grafts. AB - The treatment of aortic abdominal aneurysms with endovascular repair provides a treatment alternative for high-risk surgical candidates. Computed Tomographic Angiography (CTA) has become the preferred imaging modality for preoperative evaluation of abdominal aortic aneurysms, postoperative assessment of endovascular repair and complications, and long term surveillance. Accurate evaluation of these devices and identification of endoleaks and other complications by the radiologist is critical to assuring successful outcomes. PMID- 15115156 TI - Wegener's Granulomatosis: CT evolution of pulmonary parenchymal findings in treated disease. AB - OBJECTIVE: To determine the computed tomography (CT) evolution of various pulmonary manifestations of Wegener's Granulomatosis (WG) following appropriate pharmacologic treatment of the disease. METHODS: Eleven patients with WG were identified, each of whom had had at least two CT examinations. CTs were reviewed retrospectively to identify pulmonary lesions of WG. Lesions were categorized as nodules, cavities, lobar atelectasis, pulmonary bands, or infiltrates. To determine the evolution of each individual lesion following pharmacologic treatment, the authors compared the lesions at presentation to their appearances on follow-up CT examinations. RESULTS: A total of 112 lesions were identified (nodules = 70, cavities = 25, lobar atelectasis = 7, pulmonary bands = 6, infiltrates = 4). The mean time interval between CT examinations was 34 weeks (range: 3-248 weeks). Treated nodules tended to become smaller (33/70, 47%), to resolve (14/70, 20%), or to remain unchanged (8/70, 11%). However, the nodules became larger or cavitated in a substantial minority of cases (13/70, 19% and 2/70, 3%, respectively). Although more than half of the treated cavities became smaller (13/25, 52%) or resolved (1/25, 4%), many evolved into nodules (6/25, 24%) or enlarged (5/25, 20%). All cases of lobar atelectasis (14/14, 100%) and transpulmonary bands (6/6, 100%) were unchanged at follow-up. All infiltrates were either resolved (3/4, 75%) or substantially improved (1/4, 25%). CONCLUSION: WG has a wide spectrum of pulmonary manifestations. Nodules, cavities, and infiltrates are among the most common lesions seen on CT. Although these findings tend to improve with treatment, mixed responses are not uncommon. Lobar atelectasis and transpulmonary bands tend not to improve, even with the occurrence of clinical disease remission. PMID- 15115157 TI - Wild polioviruses: can we afford to leave them in the cold? PMID- 15115158 TI - Melioidosis, the mimicker of maladies. PMID- 15115159 TI - Elements of medical research. AB - Most medical research is empirical based on evidence rather than hunches or preferences. It follows a series of specific steps. There are no short cuts. Collection of evidence and its analysis should follow a carefully drawn protocol. Most of the modern medical research requires biostatistical tools to reach to a valid and reliable conclusion. Researcher must have an adequate knowledge and skill to be really effective. The endeavours should be consistent with the accepted medical and research ethics. Medical research can provide immense satisfaction when conducted on scientific lines, and can be occasionally frustrating when years of efforts fail to produce expected results. This article focuses on aspects that can increase the credibility of research. It is addressed to all interested in medical research, and seeking answers to questions such as what actually is research, what are its types, what specific steps should be followed, what a research protocol should contain, and what makes research credible etc. PMID- 15115160 TI - Detection of virulence attributes of Burkholderia pseudomallei. AB - BACKGROUND & OBJECTIVES: Melioidosis caused by Burkholderia pseudomallei is an emerging disease in India. This study examined the toxin activity of bacteria free culture filtrate in three different cell lines (cytotoxic assay) and its effect on Caenorhabditis elegans (nematode toxicity assay). Endotoxic activity of the viable bacteria was also studied in C. elegans (co-culture killing assay). METHODS: For toxin studies, serial doubling dilutions of unheated, heated crude and ultra filtrate of bacteria-free culture supernatants of B. pseudomallei were tested in 96-well microtitre plate containing confluent mono layers of McCoy, Hep 2 and HeLa cell lines. For the effects on C. elegans, the worms were exposed to heated and unheated bacteria-free culture supernatants in 24-well microtitre plate for 24h and then transferred to OP50 Escherichia coli lawn culture. The endotoxic activity of the live bacterium was studied by feeding the worms in the lawn culture of B. pseudomallei. RESULTS: All the clinical isolates (n=38) produced cytotoxic changes in all the cell lines. No difference was observed in the cytotoxicity of unheated, heated and ultra-filtered culture supernatant. The septicaemic isolates were observed to produce cytotoxic changes in high dilutions (1:160) of culture filtrate. None of the unheated and heated crude filtrate had deleterious effect on C. elegans, while all the live bacteria were found to be lethal to the nematode. INTERPRETATION & CONCLUSION: The culture supernatants, though produced cytopathic effect in various tissue cultures, failed to have any deleterious effect on the worms. However, live bacteria were lethal to the worms B. pseudomallei. Use of C. elegans model to detect virulence attributes of B. pseudomallei is recommended as an alternative to tissue culture methods as this can be carried out in laboratories where a tissue culture set up is not available. PMID- 15115161 TI - Effect of growth medium on hydrophobicity of Staphylococcus epidermidis. AB - The present study was undertaken to evaluate the effect of growth medium on expression of hydrophobicity of Staphylococcus epidermidis. A total of 24 hydrophobic isolates of S. epidermidis, determined by n-hexadecane adherence assay (HAA) earlier were included. Five different growth media: horse blood agar (HBA), brain heart infusion agar (BHIA), brain heart infusion broth (BHIB), tryptic soy broth (TSB) and proteose peptone broth (PPB) were used. All 24 isolates exhibited the reproducible hydrophobicity when grown on HBA; however, 20 (83.33%), 19 (79.16%), 15 (62.50%) and 13 (54.16%) isolates were found to be hydrophobic when grown in BHIA, BHIB, TSB and PPB, respectively. HBA was found to be the most suitable medium for detection of hydrophobicity of S. epidermidis followed by BHIA or BHIB. PMID- 15115162 TI - Effect of portal venous injection of donor spleen cells on skin allograft survival in rat. AB - BACKGROUND & OBJECTIVES: Pretransplantation injection of donor lymphohaemopoetic cells via portal venous route has been shown to improve allograft survival in mice. In the present study, the effect of perioperative portal venous administration of donor splenocytes on skin graft survival was investigated in comparison with intravenous administration of spleen cells in Swiss albino rat skin transplant model. METHODS: Using a single-donor survival study, skin allograft recipients received either no treatment, a single transfusion of donor spleen cells via portal vein or a single transfusion of donor splenocytes into vena cava. Spleen cell transfusion consisted 25x10(6) viable cells in a volume of 1ml given just before skin grafting. Skin graft survival was assessed by macroscopic appearance. Rejection was defined as the first day on which the entire surface of the graft was necrotic. Histologically necrosis, increased connective tissue, vascularity and polymorphonuclear leucocyte (PNL) infiltration were evaluated under light microscopy. RESULTS: In this survival study of skin allografts, with the injection of viable spleen cells into portal vein concomitant to skin grafting, significant prolongation of mean allograft survival was induced (20.3 days), compared with untreated recipients (6.5 days, P<0.001). In the histopathologic evaluation, less PNL infiltration, necrosis, increased vascularity and connective tissue repair were observed in vena porta group with no statistical significance. INTERPRETATION & CONCLUSION: It may be possible to develop protocols to induce transplantation tolerance based on the historical concept of donor specific antigen administration. However, it appears that donor spleen cell transfusion alone is not sufficient to prevent graft rejection. Thus, more efficient combination treatments are required to induce a state of durable tolerance. PMID- 15115163 TI - Mechanism of action of the cisapride-induced vasodilatation in renal vasculature of rat. AB - BACKGROUND & OBJECTIVES: Cisapride is a prokinetic agent with cholinomimetic and 5-HT4 receptor agonistic properties. It has been proposed that cisapride-induced hypotension is partly mediated by cholinergic system. The aim of this study was to investigate the mechanism of cisapride-induced dilatation in the rat isolated perfused kidney. METHODS: Left kidneys of Wistar rats were isolated and perfused via renal artery and the perfusion pressure was recorded. Cisapride given as bolus injections (10(-10)-3x10(-5) mol/l) produced dose-dependent dilatations. Perfusion of antagonists or inhibitors was started 30min before the onset of phenylephrine perfusion. RESULTS: 4-Diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP; blocker of M1, and M3 muscarinic receptors; 10(-7) mol/l) inhibited the responses to the lower doses of cisapride while, dextran (10(-7) mol/l), glibenclamide (inhibitor of ATP-sensitive potassium channels; 10(-5) mol/l) and capsaicin (for neuromediator depletion; 10(-6) mol/l) inhibited those to the higher doses. Dilatations induced by most of the doses of cisapride were inhibited by atropine (non-selective muscarinic receptor antagonist; 10(-7) mol/l), methylene blue (inhibitor of soluble guanylate cyclase; 10(-5) mol/l), 1H [1,2,4] oxadiazolo-[4,3-a] Quinoxalin-1-One (ODQ; inhibitor of soluble guanylate cyclase; 10(-5) mol/l), and NG-nitro-L-arginine (L-NOARG; NO synthase inhibitor; 10(-4) mol/l). Inhibition induced by L-NOARG was reversed by L-arginine (10(-3) mol/l). The dilatation induced by cisapride was not affected by GR113808 (5-HT4 receptor antagonist; 10(-7) mol/l) and indomethacin (cyclooxygenase inhibitor; 10(-5) mol/l). INTERPRETATION & CONCLUSION: The findings indicated that cisapride caused vasodilatation through the release of nitric oxide (NO) as a result of the release of a substance acting on muscarinic receptors, in the renal vascular bed of the rat. The role of 5-HT4 receptors and prostanoids seemed unlikely. PMID- 15115164 TI - Prediction of total body muscle mass from simple anthropometric measurements in young Indian males. AB - BACKGROUND & OBJECTIVES: The total body skeletal muscle mass plays a significant role in both health and disease states. Accurate measurement or prediction of muscle mass is useful in physiology, nutrition and clinical medicine. There are many prediction equations derived in the Western populations to estimate skeletal muscle mass, however, regression equations best fit the population they are derived from. There is hence a need to generate predictive equations for the Indian population. The objective of this study was to derive predictive equations for muscle mass from simple anthropometric measurements such as mid-arm circumference (MAC) and triceps skinfolds in a young Indian male population. METHODS: Anthropometric measurements of body weight, height, mid-arm circumference and skinfold measurements were carried out on 66 subjects with a wide range of body mass indices. Twenty four hour urine samples were collected over a 3 day period for estimating urinary creatinine excretion, from which the total body muscle mass was inferred. Linear regression was carried out between MAC and corrected arm muscle area (CAMA) with muscle mass obtained from urinary creatinine to derive a prediction equation for muscle mass. RESULTS: The prediction equation obtained for muscle mass (kg) using MAC alone was (1.641xMAC) 15.580 [r=0.72, standard error of estimate (SEE) 2.91kg] while the equation derived from CAMA alone was (0.496xCAMA)+10.183, (r=0.62, SEE=3.29kg). INTERPRETATION & CONCLUSION: A new prediction equation for the measurement of muscle mass was derived in young Indian men using simple anthropometric measurements such as mid-arm circumference and triceps skinfolds. PMID- 15115165 TI - Complementary and alternative medicines (CAM) in the management of asthma: an examination of the evidence. AB - Although individuals are using Complementary and Alternative Medical (CAM) therapies to help manage their asthma, there is no clear direction in the current guidelines for the use of CAM in asthma. This literature review undertakes to determine the current science regarding the use of CAM in asthma management. Electronic literature searched all EBM Reviews, Medline, OVID full text, and PubMed and National Complementary and Alternative Medication databases for Randomised Controlled Trials (RCT) published in English between 1997 and 2002 with keywords "asthma" and "complementary medicine" or "complementary therapy" or "alternative medicine" or "alternative therapy." Abstracts (N=197) were reviewed for inclusion in the review and duplicates discarded (N=65). Abstracts of non-RCT studies, review articles, and surveys were also discarded (N=66). Abstracts discussing environmental control measures and pharmaceutical alternatives to steroid therapy were discarded (N=9). The 15 final studies were grouped within three categories: mind-body and relaxation, manual therapies, and diet and reviewed for statistical and clinical significance, suggesting some CAM therapies have shown minimally significant improvements in asthma quality of life (breathing exercises) or pulmonary function (relaxation) and immune function (relaxation and acupuncture) in select asthma populations. Although CAM therapy is being used in the management of asthma, these 15 studies show a tendency to little or no significant difference between placebo or sham therapy. This may be due, in part, to the enhanced placebo effect of sham therapies used as control and the small size of most studies. Although the changes in the immune function seen in two studies are provocative, these changes did not translate to changes in lung function. More research is needed to assist in determining the efficacy of CAM therapies in asthma management. PMID- 15115166 TI - A comparison of two simple measures to evaluate the health status of asthmatics: the Asthma Bother Profile and the Airways Questionnaire 20. AB - Simple and concise measures for health status are desirable in clinical practice. The Asthma Bother Profile (ABP), which consists of 23 items, has been developed to assess how much asthma bothers patients. The Airways Questionnaire 20 (AQ20) is a simple instrument which consists of 20 items. The purpose of this study was to investigate how the ABP and AQ20 evaluate the health status of patients with asthma. A total of 166 patients with chronic asthma (age: 48 +/- 16 yr, 77 males) completed pulmonary function testing, measurement of airway hyperresponsiveness, dyspnea rating, assessments of their anxiety and depression (HADS; Hospital Anxiety and Depression Scale), and assessments of their health status. The health status was assessed using the ABP, AQ20, the short-form 36 health survey questionnaire (SF-36), the Living With Asthma Questionnaire (LWAQ) and the Asthma Quality of Life Questionnaire (AQLQ). The Japanese version of the ABP included only 15 'bother' items out of the original 23 items due to cultural differences. The scores on the ABP were widely distributed, whereas the scores on the AQ20 were skewed towards the milder end of the scale. The ABP had a strong correlation with the Avoidance and Distress constructs on the LWAQ, and Anxiety and Depression on the HADS (Rs = 0.56 to approximately 0.79), and its strongest correlation with the General Health (Rs = -0.64) scale among the 8 subscales on the SF-36. The AQ20 had a less significant correlation with the LWAQ, AQLQ, and SF-36 than the ABP. The ABP and AQ20 were short and simple to complete, and both measures could easily be used in clinical practice. The ABP can evaluate patients more specifically with respect to distress and bother than the AQ20. PMID- 15115167 TI - Latino children with asthma: rates and risks for medical care utilization. AB - Latino families have been reported to underutilize health care services compared with families from other ethnic backgrounds. As part of a community trial in a low income Latino population designed to decrease environmental tobacco smoke (ETS) exposure in children with asthma in San Diego, we examined unscheduled medical care for asthma. Latino families (N = 193) reported information about medical care use for their children during the past 12 months. About 23% were hospitalized, 45% used the emergency department, and 60% used urgent care services. About 8.5% of families had two or more hospitalizations in 12 months. Most families were insured by Medicaid or had no insurance. Significant risk factors for a child's hospitalization were age (under age six), failure to use a controller medication, and a parental report of the child's health status as being poor. Risk factors for emergency department use were age (under age six) and male gender. These findings indicate that low-income Latino families with young children with asthma lack the medical resources necessary for good asthma control. Quality and monitored health care with optimization of asthma management could reduce costly acute care services. PMID- 15115168 TI - Endothelial dysfunction in patients with asthma: the role of polymorphisms of ACE and endothelial NOS genes. AB - OBJECTIVES: Polymorphisms of the angiotensin converting enzyme (ACE) and endothelial nitric oxide (eNOS) genes have been implicated in asthma pathogenesis. Angiotensin II and NO have important roles in maintaining vascular tone. In this study, the relationship between endothelial dysfunction and ACE and eNOS gene polymorphisms was investigated in patients with asthma. METHODS: This cross-sectional, controlled study was conducted at the Yedikule Chest Disease Hospital and Cardiology Center in a University Hospital. Forty-nine patients with asthma (18 male, 31 female; mean age: 33+/-12 years) and 49 age- and sex-matched healthy controls (20 male, 29 female; mean age: 30+/-8 years) were included. Pulmonary function tests and flow-mediated dilatation of the brachial artery [endothelium dependent dilatation (EDD)] were examined by high-resolution ultrasonography. The ACE and eNOS genotypes were determined by PCR. RESULTS: Asthma patients showed lower EDD (12+/-6% vs. 22+/-6%, p<0.001) as compared to controls. The EDD was correlated with both predicted value of FEV1 (r=0.31, p=0.04) and predicted value of FVC (r=0.37, p=0.013). Conversely, EDD values in patients with moderate asthma were significantly lower than those in patients with mild asthma (10.1+/-5.2% vs. 14.1+/-5.7%, p=0.017). However, the ACE and eNOS genotype distribution was not significantly different between controls and asthma groups. Furthermore, EDD was not associated with both gene polymorphism of ACE and eNOS. CONCLUSION: Patients with asthma have decreased vasodilatatory response to shear stress (EDD). Decreased EDD is correlated with the severity of asthma, but not with the distribution of ACE and eNOS genotypes. PMID- 15115169 TI - Understanding pediatric inner-city asthma: an explanatory model approach. AB - Explanatory models (EMs) for asthma among inner-city school-age children and their families were examined as a means of better understanding health behaviors. Children and parents were interviewed about their concepts of asthma etiology, asthma medications, and alternative therapies. Drawings were elicited from children to understand their beliefs about asthma. Nineteen children with 17 mothers from a variety of cultural backgrounds were interviewed. Among children, contagion was the primary EM for asthma etiology (53%). Twenty-five percent of children reported fear of dying from asthma, while fear of their child dying from asthma was reported by 76% of mothers. Mothers reported a variety of EMs, some culturally specific, but the majority reported biomedical concepts of etiology, pathophysiology, and triggers. Although 76% of mothers knew the names of more than one of their children's medications, 47% thought their child's medications all had similar functions. Thirty-five percent of families used herbal treatments and 35% incorporated religion into asthma treatment. Seventy-one percent of families had discontinued medications and 23% reported currently not giving anti inflammatory medication. Reasons for discontinuing daily medications included fears of unknown side effects (53%), addiction (18%), tachyphylaxis (18%), and feeling that their child was being given too much medicine (23%). The traditional focus of asthma education is not sufficient to ensure adherence. Asthma education for children should address their views of etiology and fears about dying from asthma. Conversations with parents about their EMs and beliefs about medications and alternative therapies could assist in understanding and responding to parental concerns and choices about medications and help achieve better adherence. PMID- 15115170 TI - Serum antioxidant concentrations among U.S. adults with self-reported asthma. AB - Antioxidants may protect the lungs of people with asthma against oxidative stress. Among participants aged > or = 20 years from the Third National Health and Nutrition Examination Survey (1988-1994), we examined serum antioxidant concentrations of 771 persons with current asthma, 352 persons with former asthma, and 15,418 persons without asthma. After adjustment for age, participants with current asthma had similar mean concentrations of vitamin A, retinyl esters, vitamin C, vitamin E, vitamin E/cholesterol ratio, vitamin E/triglyceride ratio, alpha-carotene, beta-carotene, beta-cryptoxanthin, lutein/zeaxanthin, lycopene, and selenium as participants without asthma. We repeated these analyses among participants who did not use vitamin or mineral supplements. After age adjustment, participants with current asthma had lower vitamin C and beta cryptoxanthin concentrations and a lower mean vitamin E/triglyceride ratio than participants without asthma. In multiple linear regression models that included age, sex, race or ethnicity, education, smoking status, nonhigh-density lipoprotein cholesterol concentration, high-density lipoprotein cholesterol concentration, body mass index, physical activity, and alcohol use, asthma status was not significantly associated with any of the antioxidant concentrations. However, lower vitamin C concentrations were observed among people with current or former asthma than among people who never had asthma (p = 0.014). In the United States, people with asthma do not have manifest antioxidant deficiencies. PMID- 15115171 TI - Why don't children take their asthma medications? A qualitative analysis of children's perspectives on adherence. AB - Focus groups were conducted with children with asthma to generate descriptive data regarding asthma adherence. Transcripts of focus groups were analyzed using qualitative methods. Data collection occurred at an outpatient department of a university-affiliated hospital and at a summer camp for children with asthma. Thirty-six children with asthma ranging in age from 9 years, 1 month to 15 years, 3 months (M age = 11 years, 10 months) participated. Participants provided qualitative descriptions of asthma experiences, including consequences, adherence barriers, and strategies to improve adherence. Researchers also elicited participants' feedback regarding theoretically derived strategies to improve adherence. Children's perceived consequences of asthma included feeling ill, limitations on peer interactions, and medication annoyances. Frequently endorsed barriers to medication adherence were lack of motivation, difficulties remembering, and social barriers. Child-generated strategies to improve adherence included reminders, social strategies, and enhancing accessibility. When children were asked about specific strategies provided by the researchers, motivational strategies (i.e., use of reward as positive reinforcement) were the most highly endorsed. Findings revealed two paradoxes: 1) although children complained that parental reminders are annoying, they also reported that parental prompts help to improve their adherence, and 2) the use of rewards to reinforce adherence was a highly endorsed strategy when presented to children; however, children did not generate this type of strategy on their own. PMID- 15115172 TI - Knowledge of skin test results among parents of asthmatic children. AB - BACKGROUND: The control of asthma in children depends upon several factors, among which is the ability of parents to minimize the exposure of their children to specific allergens. If parents are ill-informed of the specific allergen sensitivity of their children, they may be unable to take the necessary steps to minimize exposure. OBJECTIVES: This study seeks to determine the ability of parents to recall accurately the skin test results for their children. Parents were low income, multicultural, urban residents. METHODS: One hundred eligible children with persistent asthma, between 6 and 14 years old, who were skin test positive to dust mite or cockroach allergen, were recruited from a mobile asthma clinic in Los Angeles. Caretakers were interviewed in English or Spanish. From skin test results, sensitivity and specificity of parental recall of test results were computed. The sensitivity and specificity were further stratified on demographic and exposure characteristics. RESULTS: The sensitivity was lowest for dogs (65%), but higher for all other allergens: cat 93%, roach 91%, dust mites 88%, and mold 81%. The range of specificity was from 40% to 83%. Thus, parents were more aware of positive than of negative test results. Stratification did not appreciably change the sensitivity or specificity results. CONCLUSIONS: We conclude that the sensitivity and specificity of parental response concerning skin test results is high regardless of cultural, demographic, or exposure levels of the child. PMID- 15115173 TI - Effects of a self-management asthma educational program in Taiwan based on PRECEDE-PROCEED model for parents with asthmatic children. AB - This study was conducted to evaluate the comparative effectiveness of two different asthma educational programs. One was self-management asthma education based on the PRECEDE-PROCEED model to change the influential factors based on a previous need assessment study in Taiwan. The other consisted of regular outpatient asthma education. The purposes were: 1) to compare differences in the asthma knowledge, self-efficacy, perceived effectiveness, children's cooperation, doctor-patient communication, and self-management behaviors in the experimental and control groups before education, and 2 weeks, 3 months, and 6 months after education; and 2) to compare differences in drug use, medication utilization, asthma severity, signs/symptoms of asthma, school absenteeism, and exercise ability before education, and 2 weeks, 3 months, and 6 months after education. Parents of asthmatic children were recruited from among outpatients of Chang Gung Children's Hospital, Taoyuan, Taiwan. They were grouped by registration number: those with even numbers were assigned to the experimental group, and those with odd numbers were assigned to the control group. Measurements were collected four times from all parents by means of a questionnaire and chart review. The General Linear Model: Repeat Measurement was used to compare variance differences. The following results were found. 1) Asthma knowledge, self-efficacy, perceived effectiveness, children's cooperation, and self-management behaviors significantly improved after the self-management asthma educational program based on PRECEDE-PROCEED. Except for perceived effectiveness, all variables still had good effectiveness after 6 months of follow-up. The experimental group was better than the control group in knowledge, children's cooperation, and self-management behaviors at the 3-month follow up, as well as in knowledge and children's cooperation at the 6-month follow-up. 2) In both the experimental and control groups, the educational program had a good impact on the health outcome. The average degree of drug use was reduced from 2.7 to 2.1. The number of visits was reduced from 4.75 to 3.55 per half year in the experimental group, and from 5.8 to 3.48 in the control group. The severity of asthma was reduced from 2.7 to 2.1. The signs/symptoms of asthma decreased, school absenteeism was reduced, and exercise ability improved after education at the 6-month follow-up in both groups. From the results of this study, the theory-based educational program had a good effect on self-management behaviors. From the repeat measurement analysis, we can understand the changing trend of the determinants, behaviors, and outcome indicators. The trend indicated that educational effects were sustained for at least 3 months, with some for 6 months. In order to maintain the educational effects, further specific series of educational programs can be designed based on the patterns of self-management behavior stages every 6 months. The effects of health outcomes may show significant differences using longer follow-up times in future clinical trials. PMID- 15115174 TI - Laughter-associated asthma. AB - This study was conducted to assess the prevalence of laughter-induced asthma, and to study any associations with asthma-related triggers and measures of disease activity, using a questionnaire-based survey of asthma subjects in both the community and on presentation to hospital. A total of 105 subjects participated, and 44 (41.9%) reported laughter-associated asthma. Exercise and laughter were strongly associated as triggers (p < 0.006), as well as molds and grass pollen (p = 0.03). It seems to be associated with poor asthma control as well, since hospital admissions are also more frequent in this group (p = 0.043). Laughter induced asthma is strongly associated with exercise as a trigger; the mechanism remains uncertain, but better understanding of this problem may assist in controlling difficult asthma. PMID- 15115175 TI - Allergic bronchopulmonary fungal disease caused by Saccharomyces cerevisiae. AB - We describe a patient who presented with dry cough, low-grade fever, and focal patchy shadow of pulmonary infiltrates. Remarkably, the prospective etiological agent, Saccharomyces cerevisiae was purely and repeatedly cultured from her sputum. Allergic bronchopulmonary mycosis (ABPM) was diagnosed based on clinical, serological, and pathological criteria. Although the patient described here satisfied only three of the criteria, the conclusion that the allergic bronchopulmonary disease in our case was induced by S. cerevisiae was made based on the following evidence: 1) S. cerevisiae was repeatedly isolated from the patient's sputum, 2) anti-S. cerevisiae antibody was detected in her serum, and 3) bronchoprovocation test to S. cerevisiae antigen was positive. We present here a case of allergic bronchopulmonary fungal disease caused by S. cerevisiae antigen. PMID- 15115176 TI - The impact of managed care on health care utilization among adults with asthma. AB - STUDY OBJECTIVES: To compare kinds and amounts of health care used by adults with asthma in managed care and fee-for-service settings. DESIGN: Cross-sectional structured telephone survey of Northern California adults with asthma from random samples of pulmonologists, allergist-immunologists, family practitioners, and from a random sample of the non-institutionalized population. MEASUREMENTS: Validated measures of kind of health insurance plans, kinds and amounts of services used for asthma and other reasons, demographic characteristics, severity of asthma, comorbidity, and overall health and functional status. RESULTS: Eighty one percent of the 416 adults with asthma studied were in some form of managed care (75% in HMOs and 6% in PPOs). Those in managed care (MC) and fee-for-service (FFS) did not differ substantively in the proportion with a regular source or principal provider of asthma care, with a peak flow meter or action plan, having received instructions in the use of an inhaler, reporting current use of inhaled beta-agonists, home nebulized beta-agonists, or inhaled steroids, or reporting ER visits or flu shots in the year prior to interview. Persons with asthma in MC reported significantly fewer total physician visits (after adjustment, 4.3 MC, 7.1 FFS, difference = 2.8, 95% CI -5.4, -0.1), principally because those in MC had many fewer visits to allergist-immunologists (after adjustment 4.9 MC, 21.4 FFS, difference = -16.5, 95% CI -27.8, -5.3). The two groups did not differ significantly in the proportion with asthma-related or nonasthma hospital admissions. CONCLUSIONS: Persons with asthma in fee-for-service settings reported a greater number of certain kinds of ambulatory visits, particularly visits to allergist-immunologists, for their asthma than those in managed care, but did not differ in the use of the hospital for their asthma and in nonasthma care. PMID- 15115177 TI - Two-state folding of lysozyme versus multiple-state folding of alpha-lactalbumin illustrated by the technique of disulfide scrambling. AB - The folding of lysozyme and of alpha-lactalbumin exhibits vastly different kinetics and pathways. Existing evidence indicates that folding intermediates of alphaLA form a well-populated equilibrium molten globule state that is absent in the case of hen lysozyme. We demonstrate here such divergent folding mechanisms of lysozyme and alphaLA using the technique of disulfide scrambling. Two extensively unfolded homologous isomers (beads-form) of lysozyme (Cys6-Cys30, Cys64-Cys76, Cys80-Cys94, Cys115-Cys127) and alphaLA (Cys6-Cys28, Cys61-Cys73, Cys77-Cys91, Cys111-Cys120) were allowed to refold in parallel to form the native protein. Folding kinetics was measured by the recovery of the native structure. Folding intermediates, which illustrate the folding pathway, were trapped by quenching disulfide shuffling and were analyzed by reversed-phase high-pressure liquid chromatography. The results revealed that under identical folding conditions, the folding rate of lysozyme is about 30-fold faster than that of alphaLA. Folding intermediates of lysozyme are far less heterogeneous and sparsely populated than those of alphaLA. Numerous predominant on-pathway and off pathway intermediates observed along the folding pathway of alphaLA are conspicuously absent in the case of lysozyme. The difference is most striking under fast folding conditions performed in the presence of protein disulfide isomerase. Under these conditions, folding of lysozyme undergoes a near two-state mechanism without accumulation of stable folding intermediates. PMID- 15115178 TI - Physical and kinetic properties of the family 3 beta-glucosidase from Aspergillus niger which is important for cellulose breakdown. AB - A beta-glucosidase (BGS) purified from Aspergillus niger cellulase powder (obtained from Sigma, St. Louis, MO, USA) was characterized. Electrophoresis, size exclusion chromatography, and dynamic light scattering indicated that the enzyme is a dimer of approximately 200 kDa. Five of the seven N-glycosylated oligosaccharides attached to BGS were composed of D-mannoses attached to a beta(1 4)-N-acetyl-glucosamine-beta-(1-4)-fucose-alpha-(1-6)-N-acetylglucosamine core. The other two were similar, but the cores of these did not have the D-fucose. The enzyme is a retaining glycosidase, and it also has a distinct preference for the beta-configuration at the reducing end of cellobiose. BGS is thermostable up to 65 degrees C but is sensitive to freezing and thawing. The extinction coefficient of BGS was found to be 1.8 cm(-1) mg(-1). All substrates assayed resulted in Eadie-Hofstee plots that were curved at high substrate concentrations. TLC of the reaction products showed that the substrates themselves act as acceptors when present at high concentrations. The transglucosidic activity rate is different from the hydrolytic activity rate and this causes the curvature at high substrate concentrations. The enzyme produces gentiobiose when D-glucose is the acceptor. pH optima of the Vmax(h) with pNPGlc, oNPGlc, and cellobiose were between pH 4 and 4.5, and the Km values decreased at pH values between 3 and 5. Inhibition experiments indicated that the enzyme is specific for glucosyl substrates and suggested that D-gluconolactone is a transition state analog. Studies with cello oligosaccharides and 3,4-dinitrophenyl-cellobiose showed that BGS is an exo hydrolase having at least five glucose subsites and that it cleaves from the nonreducing end. The properties of a family 3 beta-glucosidase (BG3) sequenced by Dan et al. [Dan, S., Marton, I., Dekel, M., Bravdo, B-A., He, S., Withers, S. G., and Shoseyov, O. (2000) J. Biol. Chem. 275: 4973-4980] was also studied and was shown to have very similar properties to those of BGS. Sequence analysis of a portion of BGS verified that these are the same enzymes. PMID- 15115179 TI - Regulation of NADH/CoQ oxidoreductase: do phosphorylation events affect activity? AB - We had previously suggested that phosphorylation of proteins by mitochondrial kinases regulate the activity of NADH/CoQ oxidoreductase. Initial data showed that pyruvate dehydrogenase kinase (PDK) and cAMP-dependent protein kinase A (PKA) phosphorylate mitochondrial membrane proteins. Upon phosphorylation with crude PDK, mitochondria appeared to be deficient in NADH/cytochrome c reductase activity associated with increased superoxide production. Conversely, phosphorylation by PKA resulted in increased NADH/cytochrome c reductase activity and decreased superoxide formation. Current data confirms PKA involvement in regulating Complex I activity through phosphorylation of an 18 kDa subunit. Beef heart NADH/ cytochrome c reductase activity increases to 150% of control upon incubation with PKA and ATP-gamma-S. We have cloned the four human isoforms of PDK and purified beef heart Complex I. Incubation of mitochondria with PDK isoforms and ATP did not alter Complex I activity or superoxide production. Radiolabeling of mitochondria and purified Complex I with PDK failed to reveal phosphorylated proteins. PMID- 15115180 TI - Sequence-altered peptide adopts optimum conformation for modification-dependent binding of the yeast tRNAPhe anticodon domain. AB - Amino acid contributions to protein recognition of naturally modified RNAs are not understood. Circular dichroism spectra and predictive software suggested that peptide tF2 (S1ISPW5GFSGL10 LRWSY15), selected from a phage display library to bind the modified anticodon domain of yeast tRNAPhe (ASL), adopted a beta-sheet structure. Ala residues incorporated at positions Pro4 and Gly6, both predicted to be involved in a turn, did not alter the peptide binding affinity for the ASLPhe, although major changes in the peptide's CD spectra were observed. Substitutions at three positions Pro4, Gly6, and Gly9, the latter not predicted to be in a turn, reduced the peptide's binding affinity to 4% of that of the unsubstituted tF2 and strongly influenced the peptide's secondary structure. The results suggest that peptides with different conformations, but similar affinities, adopt the optimal binding conformation, indicative of a structurally adaptive model of binding in which the modified RNA serves as a scaffold. PMID- 15115181 TI - Mapping of the antibody-binding regions on the HN-domain (residues 449-859) of botulinum neurotoxin A with antitoxin antibodies from four host species. Full profile of the continuous antigenic regions of the H-chain of botulinum neurotoxin A. AB - Previously, we mapped the antibody (Ab) and T-cell recognition regions on the HC domain (residues 855-1296) of the 848-residue heavy (H) chain of botulinum neurotoxin A (BoNT/A). We have mapped here the HN-domain (residues 449-859) regions that bind protective anti-BoNT/A Abs raised in four different species. We synthesized, purified, and characterized 29 19-residue peptides that spanned the entire HN and overlapped consecutively by 5 residues, and also region L218-231 around the L-chain's substrate-binding site. Human, horse, mouse, and chicken anti-BoNT/A Abs did not bind to the L-peptide but recognized similar HN regions within peptides 519-537/533-551/547-565/561-579 (with slight left- or right shifts), 743-761, 785-803, and 813-831/827-845 overlap. Recognition of other peptides that bound lower Ab levels showed similarities and also some differences. Peptide 463-481, strongly immunodominant with horse antisera, did not bind human, mouse, and chicken Abs. However, peptide 449-467 bound Abs in these three antisera, and the region may have shifted to the right (peptide 463 481) with horse Abs. The overlap 659-677/673-691 reacted strongly with human Abs whereas with mouse and chicken antisera, only peptide 673-691 showed low reactivity. Horse antisera had no detectable Ab binding to region(s) 659-691. The Ab-recognition regions on the H chain occupy surface locations in BoNT/A three dimensional structure, but the great part of the surface is not immunogenic. Regions recognized by the protective antisera of the four different species are prime candidates for inclusion in synthetic vaccine designs. PMID- 15115183 TI - Properties of the complex between recombinant human progastrin and ferric ions. AB - Binding of ferric ions to the hormone glycine-extended gastrin17 is essential for biological activity (Pannequin, J., et al. (2002). J. Biol. Chem. 277: 48602 48609). The aims of the current study were to determine the properties of the complex between recombinant human progastrin6-80 and ferric ions. The stoichiometry and affinity of ferric ion binding were determined by fluorescence spectroscopy. The selectivity of metal ion binding and the stability of the 59Fe(III) progastrin6-80 complex were determined by equilibrium dialysis. The stoichiometry of 2.5 +/- 0.1 moles Fe/mole progastrin, and the apparent dissociation constant of 2.2 +/- 0.1 microM, were similar to the values previously determined for glycine-extended gastrin17 at pH 4.0. Of the four trivalent and seven divalent metal ions tested, only ferrous and ferric ions bound to progastrin6-80. The ferric ion-progastrin complex was extremely stable, with a half-life of 117 +/- 8 days at pH 7.6 and 25 degrees C. We conclude that recombinant human progastrin6-80 selectively binds ferrous and ferric ions with high affinity in a stable 2:1 complex. PMID- 15115182 TI - Role of amino acid residues on the GS region of Stichopus arginine kinase and Danio creatine kinase. AB - Stichopus arginine kinase (AK) is a unique enzyme in that it evolved not from the AK gene but from the creatine kinase (CK) gene: the entire amino acid sequence is homologous with other CKs apart from the guanidine specificity region (GS region), which is identical in structure to that of AK. Ten independent mutations were introduced around the GS region in Stichopus AK. When an insertion or deletion was introduced near the GS region, the Vmax of the mutant enzyme was dramatically decreased to less than 0.1% of the wild type, suggesting that the length of the GS region is crucial for the recognition of the guanidine substrate. Replacement of Phe63 and Leu65 to Gly in the Stichopus enzyme caused a remarkable increase in the Kmarg. This indicates that Phe63 and Leu65 are associated with the arginine substrate-binding affinity. The hydrogen bond formed between the Asp62 and Arg193 residues is thought to play a key role in stabilizing the closed substrate-bound structure of AK. Mutants that eliminated this hydrogen bond had a considerably decreased Vmax, accompanied by a threefold increase in Kmarg. It is noted that the value of the Kmarg of the mutants became very close to the Kdarg value of the wild type. Six independent mutations were introduced in the GS region of Danio M-CK. Almost equivalent values of Kmcr and Kdcr in all of the mutants indicated that a typical synergism was completely lost. The results suggested that the Ile69 to Gly mutant, displaying a high Kmcr and a low Vmax, plays an important role in creatine-binding. This is consistent with the observation that in the structure of Torpedo CK, Ile69 provides a hydrophobic pocket to optimize creatine-binding. PMID- 15115184 TI - Complete amino acid sequence and location of Omp-28, an important immunogenic protein from Salmonella enterica serovar typhi. AB - Omp-28 isolated from Salmonella enterica serovar typhi presented a subunit molecular mass of 9,632 Da by MALDI-TOF MS. It was denatured, S-alkylated, and 1) directly submitted to Edman sequencing, 2) cleaved with CNBr, and 3) hydrolyzed either with endoproteinase Glu-C or Asp-N. The major CNBr peptide containing the C-terminal portion of Omp-28 was isolated by tricine-SDS-PAGE and electroblotted whereas Omp-28 enzymatic peptides were isolated by C18-RP-HPLC. All peptides were sequenced. This approach allowed the elucidation of the complete primary structure of Omp-28. Its amino acid sequence is identical to that deduced from part of the DNA of the "putative periplasmic transport protein" of either S. enterica serovar typhimurium and a multiple drug resistant S. enterica serovar typhi. Omp-28 homologous protein sequences were also deduced from Escherichia coli and Yersinia pestis genomic DNA. All proteins had their secondary structures predicted. Immunogold cytochemistry indicated that Omp-28 is found on the bacterium outer membrane. PMID- 15115185 TI - Detection of tryptophan to tryptophan energy transfer in proteins. AB - Forster resonance energy transfer (FRET) studies usually involve observation of intensity or lifetime changes in the donor or acceptor molecule and usually these donor and acceptor molecules differ (heterotransfer). The use of polarization to monitor FRET is far less common, although it was one of the first methods utilized. In 1960, Weber demonstrated that homotransfer between tryptophan molecules contributes to depolarization. He also discovered that the efficiency of homotransfer becomes much less effective upon excitation near the red-edge of the absorption. This "red-edge effect" was shown to be a general phenomenon of homotransfer. We have utilized Weber's red-edge effect to study tryptophan homotransfer in proteins. Specifically, we determined the polarization of the tryptophan fluorescence upon excitation at 295 nm and 310 nm (near the red-edge). Rotational diffusion leads to depolarization of the emission excited at either 295 nm or 310 nm, but homotransfer only contributes to depolarization upon excitation at 295 nm. Hence, the 310/295 polarization ratio gives an indication of tryptophan to tryptophan energy transfer. In single tryptophan systems, the 310/295 ratios are generally below 2 whereas in multi-tryptophan systems, the 310/295 ratios can be greater than 3. PMID- 15115186 TI - Activation parameters for the spontaneous and pressure-induced phases of the dissociation of single-ring GroEL (SR1) chaperonin. AB - We investigated the dissociation of single-ring heptameric GroEL (SR1) by high hydrostatic pressure in the range 0.5-3.0 kbar. The kinetics were studied as a function of temperature in the range 15-35 degrees C. The dissociation processes at each pressure and temperature showed biphasic behavior. The slower rate (k1,obs) was confirmed to be the self-dissociation of SR1 at any specific temperature at atmospheric pressure. This dissociation was pressure independent and followed concentration-dependent first-order kinetics. The self-dissociation rates followed normal Eyring plots (In k1,obs/T vs. 1/T) from which the free energy of activation (deltaG++ = 22 +/- 0.3 kcal mol(-1)), enthalpy of activation (deltaH++ = 18 +/- 0.5 kcal mol(-1)), and entropy of activation (deltaS++ = -15 +/- 1 kcal mol(-1)) were evaluated. The effect of pressure on the dissociation rates resulted in nonlinear behavior (ln k2,obs vs. pressure) at all the temperatures studied indicating that the activation volumes were pressure dependent. Activation volumes at zero pressure (V++o) and compressibility factors (beta++) for the dissociation rates at the specific temperatures were calculated. This is the first systematic study where the self-dissociation of an oligomeric chaperonin as well as its activation parameters are reported. PMID- 15115188 TI - Erectile dysfunction and cardiovascular considerations: managing patients effectively with phosphodiesterase type 5 inhibitors. PMID- 15115187 TI - Conformation-dependent interaction of alpha-lactalbumin with model and biological membranes: a spin-label ESR study. AB - Alpha-lactalbumin (alpha-LA) is biosynthesized and stored at the smooth endoplasmic reticulum (ER), then transferred to the Golgi lumen when prolactin stimulation of lactose biosynthesis and secretion takes place. Because both environments are composed of membranes, it was of interest to examine the interactions of alpha-LA with relevant model and biological membranes. Using the ESR spin-labeled fatty acid analog 5-doxyl stearic acid, we found evidence reflecting the insertion of "acid-shocked" molten globule (MG) alpha-LA into lecithin or phosphatidylserine (PS) multi-lamellar vesicles. An additional approximately 3 G immobilization was observed in the alpha-LA-lecithin sample versus the lipid alone. With PS, the increased immobilization was almost 6 G, reflecting an enhanced effect caused by strong electrostatic interactions between the positively charged protein with the negatively charged headgroup at pH 2.4. This was also reflected in the broadening of the PS:alpha-LA phase transition. Additionally, we have demonstrated that alpha-LA in its apo-form also shows similar insertion characteristics with both model and natural lipid membranes. Upon addition of calcium, the apo-form is released from the membrane as the Ca(2+)-bound protein. PMID- 15115189 TI - Role of the cardiologist: clinical aspects of managing erectile dysfunction. AB - Erectile dysfunction (ED) is often a marker for serious underlying cardiovascular disease (CVD), and cardiologists are increasingly involved in the care of men with ED. It is important to ask specifically about ED when evaluating men with CVD, since they may be embarrassed to volunteer this information. During the clinical workup, it is also important to check for contributing factors to ED such as diabetes, depression, stress, alcohol abuse, and cardiovascular risk factors. Patients should be advised that many treatment options are available for ED, including the phosphodiesterase type 5 (PDE5) inhibitors. The PDE5 inhibitors are safe and effective in most patients with CVD, including those taking multiple antihypertensive drugs. Furthermore, they have no deleterious effect on exercise capacity, heart rate, or extent of exercise-induced ischemia. In the future, the PDE5 inhibitors may have a role in reducing pulmonary hypertension in persons with primary pulmonary arterial hypertension (PAH) or congestive heart failure. The one major precaution for men taking PDE5 inhibitors is to avoid concomitant administration of therapeutic and recreational nitrate preparations. Patients with chest pain suggestive of a heart attack need to inform emergency room (ER) personnel if they are taking a PDE5 inhibitor. Similarly, before giving nitrates, ER personnel need to ask patients if they have used PDE5 inhibitors. Nitrates should not be given for at least 24 h after a patient uses sildenafil or vardenafil and at least 48 h after a patient uses tadalafil. PMID- 15115190 TI - Cardiovascular risk stratification and cardiovascular risk factors associated with erectile dysfunction: assessing cardiovascular risk in men with erectile dysfunction. AB - Cardiovascular risk factors are known to be associated with the presence of erectile dysfunction (ED), and ED is more common in men with cardiovascular disease (CVD). The Princeton Guidelines provide a strategy for assessing cardiac risk and planning a safe return to sexual activity. Men at low risk, who have fewer than three cardiac risk factors, can resume intercourse with very little concern for an untoward cardiac event. Men at high risk need to have their primary disease controlled, and should be stabilized before planning to resume sexual activity. Prior to resuming intercourse, men in the indeterminate category are candidates for restratification as either low or high risk. Certain cardiovascular risk factors may be predictors of ED: most notably, hyperlipidemia, hypertension, and diabetes. Advancing age is also a strong predictor of ED. The close correlation between ED and CVD is a signal to evaluate the sexual histories of all men who present for cardiovascular evaluation and, perhaps, of all men in general. Early detection of ED may allow for early diagnosis and management of CVD and diabetes. PMID- 15115191 TI - Phosphodiesterase type 5 inhibitor differentiation based on selectivity, pharmacokinetic, and efficacy profiles. AB - The mechanism of action of the phosphodiesterase type 5 (PDE5) inhibitors (i.e., sildenafil, tadalafil, and vardenafil) involves inhibition of the PDE5 isoenzyme located in penile vascular smooth muscle cells. Sexual stimulation triggers the release of nitric oxide (NO), stimulating the release of guanylyl cyclase, leading to an increase in intracellular cyclic guanosine monophosphate (cGMP) concentrations, a decrease in intracellular calcium, and ultimately relaxation of the vascular smooth muscle in the corpus cavernosum and penile erection. The PDE5 inhibitors have no effect on the penis in the absence of sexual stimulation. Although the various PDE5 inhibitors differ with respect to selectivity and pharmacokinetic profiles, efficacy and safety of these agents are comparable in broad populations of men with erectile dysfunction (ED), including those with diabetes or those taking multiple antihypertensive agents. The most frequently reported adverse events of the PDE5 inhibitors are related to their mild vasodilatory effects and include headache, flushing, dyspepsia, and nasal congestion or rhinitis. Side effects are generally reversible and tend to diminish during continued treatment. Differences in pharmacokinetic properties among the PDE5 inhibitors include the fact that sildenafil and vardenafil have a shorter duration of action (approximately 4 h) compared with the longer period of responsiveness observed with tadalafil (up to 36 h). In addition, in the presence of high-fat food, absorption of sildenafil and vardenafil may be delayed; however, the rate and extent of tadalafil absorption are unaffected by high-fat food. PMID- 15115192 TI - Novel phosphodiesterase type 5 inhibitors: assessing hemodynamic effects and safety parameters. AB - The relaxing effect of the phosphodiesterase type 5 (PDE5) inhibitors on vascular smooth muscle has attracted much attention, especially in persons with cardiovascular disease. The results of early studies showed that sildenafil slightly reduces systolic and diastolic blood pressures and has no effect on heart rate, while being safe and well tolerated. Studies also indicate that sildenafil does not contribute to the development of myocardial infarction or ischemia. Similar benign effects on hemodynamics and cardiac events have also been demonstrated for tadalafil and vardenafil. None of the PDE5 inhibitors adversely affects total exercise time or time to ischemia during exercise testing in men with stable angina. It is key to avoid concomitant administration of nitrates with any of the PDE5 inhibitors, because this combination can cause increased vasodilation and a subsequent drop in blood pressure. Sildenafil has an alpha-blocker precaution; tadalafil is contraindicated with alpha blockers except for 0.4 mg tamsulosin; vardenafil is contraindicated with alpha blockers. PMID- 15115193 TI - An assessment for the development of a national surveillance system for travel acquired enteric disease. PMID- 15115194 TI - Poultry breeding in the post-genomics era. AB - 1. I introduce the problems facing poultry breeding in the future and the role biotechnology may have to play. 2. I then review recent research in the genomics and bioinformatics of poultry and where this may lead in the near future. 3. Finally, I discuss the scientific and commercial opportunities of this research and strategies for implementation into poultry breeding practices. PMID- 15115195 TI - Different concentrations and sources of dietary fibre may improve the welfare of female broiler breeders. AB - 1. Broiler breeder females were fed restricted allocations of food to meet target body weights. They were fed on a standard wheat-soy ration or on one of three series of rations containing 50, 100 or 200 g/kg of ground oat hulls, ground unmolassed sugar beet pulp or sunflower meal. Changes in behaviour and heterophil lymphocyte ratio (HLR) were measured at 5, 10 and 15 weeks of age. 2. Body weight at 15 weeks of age decreased in a linear manner with increasing concentrations of the experimental ingredients. 3. Decreased spot pecking was observed in birds fed on the rations containing 50 g/kg of sugar beet pulp and 200 g/kg of oat hulls. 4. The HLR was higher in birds fed on the control diet and diets containing 50 g/kg of the high fibre ingredients. 5. Rations containing sugar beet pulp were associated with higher water contents in the gastrointestinal tract and it is proposed that this improved satiety and welfare. PMID- 15115196 TI - Pop hole passages and welfare in furnished cages for laying hens. AB - 1. This study included two designs of furnished cages for 16 hens; H-cages divided into two apartments by a partition with pop holes in the middle of the cage, and fully open O-cages, without a partition. The hypothesis was that in this rather large group of birds the pop hole partition would benefit the birds by allowing them to avoid or escape from potential cannibals, feather-peckers or aggressive hens. All cages had two nests, two perches and one litter box. 2. A total of 10 cages (5 H and 5 O) were stocked with Lohmann Selected Leghorn (LSL) and 8 cages (4 H and 4 O) with Hy-Line W36. No birds were beak-trimmed. 3. Heterophil/lymphocyte (H/L) ratios, duration of tonic immobility (TI) and exterior appearance (scoring of plumage condition and wounds at comb or around cloaca) were used as indicators of well-being. Total mortality and deaths due to cannibalism were also recorded. 4. Visits to nests and passages through partition pop holes were studied in samples of 35 and 21 birds, respectively, using a technique based on passive integrated transponder (PIT) tags. 5. Cage design (H- vs O-cage) had no effect on the welfare traits chosen. 6. Hy-Line birds showed higher H/L ratios, longer duration of TI and better plumage condition than LSL birds. These differences are discussed in terms of stress thresholds and copying strategies. 7. On days when a hen made visits to nests, the visiting frequency was 1.4 and the total time in the nest was 41 min on average. Hens made use of the pop hole passages between 1 and 8 times per hen and day. 8. Overall low levels of aggression, lack of injuries or deaths due to cannibalism, and plumage condition indicating moderate feather pecking, together imply a low need to escape. The pop holes were used frequently and birds distributed well between compartments showing that the system worked well. However, at this group size there was no evidence in the measured traits that H-cages provided a better housing environment. PMID- 15115197 TI - Effect of one or two pre-pubertal long days on age at first egg in domestic pullets. AB - 1. Two trials were conducted, using 288 brown-egg hybrid pullets in each, to determine the effect on age at first egg (AFE) of exposure to one or two 'long days' during the rearing period. In the first trial, birds were given a single 'long day' of 10, 12 or 14 h at 75, 89 or 103 d of age, with controls maintained on 8-h photoperiods. All treatment groups were transferred to cages at 110 d, and half the birds from each treatment combination given a 6-h increment in photoperiod at 116 d, with the remainder held on 8-h photoperiods. In the other trial, birds were given one or two long days of 14 or 16 h at 96, 107, 117 or 128 d of age; controls were again held on 8-h photoperiods. All groups were moved to cages at 130 d but maintained on 8-h photoperiods. 2. AFE was not significantly affected by one or two pre-pubertal long days, irrespective of when the long day was given or its length. AFE was advanced by 2 to 3 weeks following a transfer from 8 to 14 h at 116 d, independently of whether or not the birds had received a previous long day exposure. 3. It appears that a full-fed egg-type hybrid requires more than two cycles of long days to initiate rapid gonadal development, and that exposure to a single long day during the rearing period will have minimal effect on the timing of sexual maturation and no influence on the response to a subsequent permanent transfer to long days. PMID- 15115198 TI - Developmental states of the collagen content, distribution and architecture in the pectoralis, iliotibialis lateralis and puboischiofemoralis muscles of male Red Cornish x New Hampshire and normal broilers. AB - 1. Developmental states of the collagen content, distribution and architecture in the pectoralis (PT), iliotibialis lateralis (ITL) and puboischiofemoralis (PIF) muscles of male Red Cornish x New Hampshire (RN, 80 d, body weight 2.9 kg) and normal (3.1 kg) broilers were evaluated. 2. In PT muscle the total amount of collagen was significantly greater in RN broilers (3.33 mg/g) than in normal ones (1.71 mg/g). This higher collagen content in RN broilers was based mainly on the closer mesh sizes of endomysial honeycomb. The collagen structures in the perimysia also differed between broiler types, when more collagen fibres were observed in RN broilers. 3. ITL muscle contained total collagen of 4.10 to 5.00 mg/g. Types I and III collagens were distributed on the perimysia at higher percentages in RN broilers (31.6%, 37.2%) than normal (15.6%, 30.8%), respectively. The thick bands of tough collagen fibres characteristic of ITL muscle perimysium in cockerels had not yet developed in these broilers. 4. Total collagen was 4.63 to 6.29 mg/g in PIF material with fascia. In PIF muscle the perimysial collagen fibres had not yet attained their full growth but consisted of densely packed fibrils. PIF muscle was characterised by the earlier maturing collagen structure. 5. These results show that a perimysial collagen structure in broilers is still in an undeveloped state. It is supposed that tenderness of broiler meat is attributed mainly to characteristics of the collagen distribution, in which the majority of types I and III collagens is distributed on the closer mesh of endomysial honeycomb. PMID- 15115199 TI - Morphology of the small intestinal mucosal surface of broilers in relation to age, diet formulation, small intestinal microflora and performance. AB - 1. Three experiments were performed to relate morphological characteristics of the small intestinal mucosal surface to age, dietary factors, small intenstinal microflora and performance of broilers. Characterisation of the small intestinal mucosal surface using a dissecting microscope was based on the orientation of the villi, villus shape and the presence of convoluted villi. 2. In Trial 1, the morphological changes of the mucosal surface were studied weekly in the period from 7 to 28 d of age. At d 7 mainly tongue- and leaf-shaped villi together with some ridge-shaped ones were observed in the middle section of the small intestine, displaying a regular zigzag pattern on 53% of the mucosal surface. During the period from d 7 to 14, the area with ridge-shaped villi increased from 7 to 63% and did not change significantly over the next 2 weeks. 3. In Trial 2, three protein sources, soy isolate (SI), wheat gluten (WG), hydrolysed wheat gluten (HWG) and SI with added L-glutamine (SI + Gln), were studied with respect to their effect as dietary components on villus morphology in the mid-small intestine and performance. Diets were fed with (0 to 14 d) and without pectin (14 to 21 d). Feed conversion ratio on the HWG diet improved in comparison to the native WG diet. During the period 0 to 14 d of age the mucosal area with zigzag oriented villi increased when the pectin diet was supplemented with Gln. Moreover, weight gain of birds fed the SI + Gln diet increased in the period 41 to 21 d. 4. In Trial 3, a study was made of the morphological response of the villi to a stimulation of microbial activity in the digesta after addition of highly methylated pectin to the soybean meal (SBM) diet. This was performed with and without inoculation of a non-virulent Salmonella typhimurium on d 7. By d 21 the birds fed the pectin diet showed impaired weight gain and higher feed conversion. The pectin affected the mucosal surface by decreasing the area with the zigzag pattern and increasing the area with convoluted, mainly ridge-shaped villi. The Salmonella typhimurium infection increased the effects of pectin on performance and mucosal morphology. PMID- 15115200 TI - Influence of bird strain on competitive exclusion of Campylobacter jejuni in young chicks. AB - 1. Newly hatched chicks of either layer or broiler strain were treated orally at regular intervals with either homologous or heterologous gut-flora preparations from young donor birds, in an attempt to prevent subsequent colonisation with Campylobacter jejuni by 'competitive exclusion' (CE). 2. Donors of 3 to 10 d of age were chosen to correspond with the period in which intensively reared poultry are least likely to become colonised with Campylobacter. 3. In two separate trials, material from donor layer hens (ISA Brown) protected male chicks of the same strain against a low (195 to 360 cfu/bird) Campylobacter challenge, but the same kind of material was ineffective when administered to chicks of a broiler strain (JA957). 4. Two further trials involved treatment preparations from young broilers, which failed to prevent Campylobacter colonisation of broiler chicks, even when colonisation occurred relatively slowly from a challenge of 90 to 94 cfu/bird. 5. It was concluded that any CE effect observed was strongly dependent on bird strain. PMID- 15115201 TI - Causes for improvement in nutritive value of broiler chicken diets with whole wheat instead of ground wheat. AB - 1. Two experiments were conducted with broiler chicks to compare nutritive values of ground and whole wheat diets containing titanium oxide as indigestible marker. 2. In Experiment 1, a wheat and soy isolate based cold-pelleted diet, in which all the wheat was ground in a hammer mill to pass through a 3 mm sieve, was compared with a diet in which about half (375 g/kg) of the ground wheat was taken out of the diet and supplied as whole wheat mixed into the diet after pelleting. 3. In Experiment 2, a diet based on ground wheat and soy isolate which was pelleted after preconditioning (final temperature 81 degrees C) was compared with a diet in which 500 g of the ground wheat was replaced by whole wheat mixed into the diet before pelleting. 4. In Experiment 1, replacement of ground wheat with whole wheat increased apparent metabolisable energy (AME) content and starch digestibility measured at ileal as well as faecal level, while weight gain and feed conversion efficiency (FCE) were not affected. Jejunal chyme showed increased amylase activity and bile salt concentration. 5. In Experiment 2, replacement of ground wheat with whole wheat increased dry matter content as well as bile content of gizzard, and stimulated an increase in gizzard as well as pancreas weight. Jejunal amylase and bile concentrations were not significantly affected and neither were growth, FCE or AME. 6. Results from this experiment indicate that the improved feed value sometimes observed with whole wheat may be associated with modulation of digestive processes resulting in increased pancreas and liver secretions. PMID- 15115202 TI - Defaecation and weight of the gastrointestinal tract contents after feed and water withdrawal in broilers. AB - 1. Broilers were held under constant light for up to 24 h with access to feed and water, access to water only, or access to neither feed nor water. 2. Birds given continuous access to feed ate evenly over the 24-h period. Feed deprivation reduced drinking behaviour to about half of that when feed was available. Deprivation of feed, or of feed and water, had similar effects on defaecation and the reduction in the weight of the gut contents. It reduced the overall number of defaecations over the 24-h period to about 40% of that in birds given food and water, and the weight of the contents by about 75%. The maximum effect of deprivation on defaecation was seen after about 4 h, and on gut contents weight after about 12 h. 3. With longer fasting the contents of most parts of the gut, except the caeca, became wetter. 4. The findings may have implications for the practice of preslaughter food withdrawal to reduce the potential for soiling of the live birds and contamination of the carcase after slaughter. However, the likelihood is that relatively short overall feed deprivation times of not more than 8 to 12 h before killing may be desirable, but specifying firm recommendations on optimal feed withdrawal times requires further work. PMID- 15115203 TI - Dietary isoleucine responses in male broiler chickens. AB - 1. Three experiments were conducted to measure growth and carcase responses of growing and finishing broilers fed on test diets formulated to be deficient in isoleucine (Ile). 2. Dose titration methodology was used to measure growth and carcase responses of growing and finishing broilers to graduations of Ile in three additional experiments. 3. The experiments were conducted from d 18 to 30, 30 to 42, and 42 to 56. 4. Broilers given Ile-deficient test diets had poorer weight gain, feed conversion and carcase responses than broilers fed on Ile test diets containing a surfeit of Ile. Adding supplemental Ile to the test diet resulted in equivalent growth and carcase responses to those of broilers fed on the control diet with equal Ile from intact protein sources. 5. Recommended total Ile needs varied between 6.7 and 7.1 g/kg from d 18 to 30, 6.4 to 6.6 g/kg from d 30 to 42, and 5.5 to 6.6 g/kg from d 42 to 56. PMID- 15115204 TI - Influence of phytase and xylanase, individually or in combination, on performance, apparent metabolisable energy, digestive tract measurements and gut morphology in broilers fed wheat-based diets containing adequate level of phosphorus. AB - 1. The aim of the present study was to examine the influence of microbial phytase and xylanase, individually or in combination, on performance, apparent metabolisable energy, digesta viscosity, digestive tract measurements and gut morphology in broilers fed on wheat-soy diets containing adequate phosphorus (P). The wheat-soy basal diet was formulated to contain 4.5 g/kg non-phytate P and the experimental diets were formulated by supplementing the basal diet with xylanase (1000 xylanase units/kg diet), phytase (500 phytase units/kg diet) or a combination of phytase and xylanase. 2. Supplemental phytase improved the weight gains and feed efficiency by 17.5 and 2.9%, respectively. Corresponding improvements due to the addition of xylanase were 16.5 and 4.9%, respectively. The combination of phytase and xylanase caused no further improvements in broiler performance. 3. Individual additions of xylanase or phytase resulted in numerical improvements in apparent metabolisable energy (AME), but the differences were not significant. The combination of the two enzymes significantly increased AME. Addition of xylanase and the combination of the two enzymes reduced the viscosity of digesta in all sections of the intestine. Phytase supplementation reduced digesta viscosity in the duodenum and ileum, but not in the jejunum. 4. Enzyme supplementation lowered the relative weight and length of the small intestine. Additions of xylanase and phytase reduced the relative weight of the small intestine by 15.5 and 11.4%, respectively, while the corresponding reductions in the relative length of the small intestine were 16.5 and 14.1%, respectively. The combination of phytase and xylanase had no further effects on the relative weight and length of the small intestine compared with the xylanase group. 5. The addition of phytase increased villus height in the duodenum and decreased the number of goblet cells in the jejunum compared with those on the unsupplemented basal diet. Xylanase supplementation tended to increase goblet cell numbers in the duodenum and decreased crypt depth in thejejunum. The combination of phytase and xylanase increased villus height in the ileum and crypt depth in thejejunum and ileum. 6. In summary, the present results showed that the addition of a microbial phytase, produced by solid state fermentation and containing significant activities of beta-glucanase and xylanase, was as effective as xylanase in improving the performance of broiler chickens fed on wheat-based diets containing adequate levels of P. Improved performance with enzyme supplementation was generally associated with reduced digesta viscosity, increased AME, and reduced relative weight and length of small intestine. PMID- 15115205 TI - Linear regression approach to study amino acid digestibility in broiler chickens. AB - 1. An experiment was conducted to investigate whether a linear regression approach is a suitable tool for determining the amino acid (AA) digestibility up to the terminal ileum of broiler chickens. Solvent-extracted rapeseed meal (RSM) was used as the model ingredient. 2. Ten diets with 5 different inclusion rates of RSM (60, 120, 180, 240 and 300 g/kg, corresponding to crude protein concentrations from 170 to 250 g/kg in the diet), each without or with a supplementation of phytase (500 U/kg), were fed ad libitum to broiler chickens between 14 and 21 d of age. Seven pens of 12 chickens were allocated to each treatment. Digesta were sampled on a pen basis from the section of the gastrointestinal tract between Meckel's diverticulum and 2 cm anterior to the ileo-caeco-colonic junction. Titanium dioxide was included as an indigestible marker. 3. The amounts of crude protein and AAs digested up to the terminal ileum constantly increased with increasing AA intake over the entire range of intakes. When the amount of an AA digested at the terminal ileum is linearly regressed against its intake, the deviation of the slope from 1 is caused by both the unabsorbed AA from RSM and from specific endogenous losses related to RSM. These slopes varied between 0.68 and 0.88 for individual AAs, and the slopes were unaffected by phytase supplementation. 4. It is suggested that a linear regression approach be adopted to study the AA digestibility of raw materials in chickens. Digestibility determined this way does not need any correction for basal endogenous loss. PMID- 15115206 TI - Chick responses to dietary arginine and methionine levels at different environmental temperatures. AB - 1. Two experiments were conducted with broiler chicks in battery brooders from 1 to 21 d to determine the broiler chicks' responses to arginine (Arg) and methionine (Met) combinations at control (22 to 25 degrees C) and warm (32 to 35 degrees C) temperatures. 2. In Experiment 1, two levels of Arg (15.2 and 25.2 g/kg of the diet) and two levels of Met (3.5 and 5.5 g/kg) of a maize-soy based diet were fed at two temperatures, 22 or 32 degrees C. Results of Experiment 1 were similar to those of Experiment 2, but most treatment differences were not significant. 3. In Experiment 2, chicks were randomly allotted to 9 dietary treatments: 3 levels of Arg (15.2, 25.2 and 35.2 g/kg of the diet) x 3 levels of Met (3.5, 5.5 and 7.5 g/kg of the diet) at 25 or 35 degrees C. At the warmer temperature, chick growth depression from supplemental Arg was not as severe as at the control temperature (significant Arg x temperature interaction); neither were growth increases as large from supplemental Met (significant Met x temperature interaction). 4. Kidney and breast muscles were collected for arginase activity and creatine analysis, respectively. Remaining chicks were fasted for 10 h and re-fed. Excreta from the next 24 h were collected for total creatine and creatinine analysis. There were no effects of either Arg or Met on muscle creatine concentration at either control or warm temperatures. Chicks raised at 25 degrees C excreted more creatine and creatinine than those raised at 35 degrees C. 5. These results confirm that temperature affects responses to dietary Arg and Met and suggest that the higher temperature slowed the Arg metabolism of chicks through the creatine synthesis pathway. PMID- 15115207 TI - The effects of phytase and phytic acid on the loss of endogenous amino acids and minerals from broiler chickens. AB - 1. The effects of myo-inositol hexaphosphate (IP6) and phytase (EC 3.1.3.26) on the excretion of endogenous compounds were investigated using growing broiler chickens. 2. A total of 32 female Ross broilers were used in a precision feeding assay involving a 2 x 2 factorial arrangement of treatments. The materials administered were glucose, glucose + 1000 units of phytase activity (FTU), glucose + 1 g of IP6 and glucose + 1 g of IP6 + 1000 FTU. Excreta were collected quantitatively over a 48-h period following intubation of the test materials. The excretion of nitrogen, amino acids, minerals, sialic acid and phytate phosphorus was determined. 3. The ingestion of 1 g of IP6 by broilers increased the excretion of endogenous nitrogen, amino acids, iron, sodium, sulphur and sialic acid compared with birds fed on glucose. Supplementation of IP6 with exogenous phytase reduced the excretion of endogenous amino acids, calcium, sodium, phytate phosphorus and sialic acid compared with birds fed IP6. 4. It can be concluded that IP6 increases the excretion of endogenous minerals and amino acids in broiler chickens. Part of the beneficial effects of the addition of exogenous phytases to the diets of poultry appears to be mediated through a reduction in endogenous losses of these nutrients. PMID- 15115208 TI - Preen gland function in layer fowls: factors affecting preen oil fatty acid composition. AB - 1. Preen oil derived from the preen gland has previously been shown to differ in its composition between birds of different ages, sex and diet. As a part of a larger study on preening behaviour and its components, the relative percentages of fatty acids in preen oil were examined in laying hens that differed in age, beak trimmed status, feather pecking status (pecker and pecked) and method of sampling (either direct from the preen gland or from lipid extracted from feathers). 2. Five experiments are described. Fatty acids extracted from lipid were analysed by gas chromatography (GC). Fatty acid composition was affected by age, by whether or not a bird was feather pecked (but not if it was a feather pecker) and by lipid source. 3. Changes in preen oil composition with age (shown here) may be due to changes in circulating concentrations of hormones. Differences in preen oil composition between feather pecked and non-pecked birds may influence plumage odour and therefore taste, making the plumage of some birds more attractive to pecking than others. 4. The proportions of some fatty acids derived from feathers differed to those extracted from the preen gland. Lipid found on feathers is most likely a combination of that from the preen gland and from sebaceous secretions from the skin. PMID- 15115209 TI - Ascorbic acid and melatonin reduce heat-induced performance inhibition and oxidative stress in Japanese quails. AB - 1. The effects of ascorbic acid (L-ascorbic acid) and melatonin supplementation on performance, carcase characteristics, malondialdehyde (MDA) as lipid peroxidation indicator, ascorbic acid, retinol, tocopherol and mineral status in the Japanese quail (Coturnix coturnix japonica) exposed to high ambient temperature were evaluated. 2. Two hundred and forty Japanese quails (10 d old) were randomly assigned to 8 treatment groups consisting of 10 replicates of three birds each. The birds were kept in a temperature-controlled room at 22 degrees C (Thermoneutral, TN groups) or 34 degrees C (for 8 h/d; 09:00 to 17:00 h; Heat stress, HS groups). Birds in both TN and HS were fed either a basal (control) diet or the basal diet supplemented with 250 mg of L-ascorbic acid/kg of diet (Ascorbic acid group), 40 mg of melatonin/kg of diet (Melatonin group) or both (Ascorbic acid + Melatonin group). 3. Supplementing heat-stressed quails with ascorbic acid and melatonin improved performance compared with the control group. Effects generally were greatest in quails supplemented with both ascorbic acid and melatonin. 4. Although supplementation did not consistently restore the concentrations of serum ascorbic acid, retinol and tocopherol to those of TN groups, these concentrations increased significantly with supplementation. Furthermore, serum and liver MDA and serum cholesterol and glucose concentrations were lower in the supplemented groups than in the heat-stressed controls. 5. Within each environment, excretion of Ca, P, Mg, Zn, Fe and Cr were lowest in the combination group and, in all cases, highest in the HS group. Interactions between diet and temperature were detected for live weight gain, cold carcase weight, MDA, ascorbic acid, tocopherol concentrations and excretion of zinc. 6. The results of the study indicate that ascorbic acid and melatonin supplementation attenuate the decline in performance and antioxidant and mineral status caused by heat stress and such supplementation may offer protection against heat-stress-related depression in performance of Japanese quails. PMID- 15115210 TI - Changes in light intensity can influence age at sexual maturity in domestic pullets. AB - 1. Shaver White and ISA Brown pullets were reared to 140 d in groups of 8 in cages on a 10-h photoperiod of incandescent light and maintained at an illuminance of 3 or 25 lux, or transferred from 3 to 25 lux or from 25 to 3 lux at 63 or 112 d of age. 2. There was no significant difference in sexual maturity, measured as eggs per 100 bird.d at 139 and 140 d, for ISA Brown maintained on 3 or 25 lux, but Shaver White pullets exposed to constant 3 lux matured significantly later than those maintained on 25 lux. 3. In Shaver Whites, sexual maturity was significantly delayed by an increase from 3 to 25 lux at 63 and 112 d, and advanced by a decrease from 25 to 3 lux at 112 d. Sexual maturity of ISA Browns was not significantly affected by a change in illuminance at 63 or 112 d, though responses were in the same direction as for Shaver Whites. 4. In both breeds, total feed consumed to 112 d was higher for birds on 3 lux than 25 lux, but lower between 112 d and 140 d when birds on 25 lux underwent rapid sexual development. In both breeds, body weight at 63 d was higher for birds exposed to 3 lux than 25 lux, but body weight gain thereafter was similar for the two light intensities. 5. In both breeds, plasma luteinising hormone (LH) concentration at 63 and 112 d was lower in birds maintained on 3 lux than 25 lux. At 63 and 112 d, transfers from 25 to 3 lux depressed, whereas transfers from 3 to 25 lux at 63 d, but not at 112 d, increased plasma LH. 6. Advances or delays in sexual maturity induced by changes in illuminance were not correlated with differences in feed intake, body weight gain, or with changes in plasma LH. 7. One possible explanation for the inverse relationship between the direction of change in illuminance at 63 and 112 d in pullets exposed to a 10-h photoperiod and the age at which they became sexually mature is that changes in light intensity and/or spectral composition affect the entrainment of the circadian rhythm of photoinducibility, to effect a phase shift in the photoinducible phase and/or the responsiveness of phototransduction pathways. PMID- 15115211 TI - Uptake and distribution of astaxanthin in several tissues and plasma lipoproteins in male broiler chickens fed a yeast (Phaffia rhodozyma) with a high concentration of astaxanthin. AB - 1. The experiments were conducted to evaluate astaxanthin (Ax) uptake in several tissues and plasma lipoproteins of male broiler chickens fed on Phaffia rhodozyma containing a high concentration of Ax. 2. Male broiler chicks (5 weeks of age) fasted for 16h were given 0 or 45 mg Ax as Phaffia rhodozyma through the crop and blood was collected over the following 24 h. Ax appeared in the plasma at 2 h after administration into the crop. Most (more than 70%) of the Ax was contained in the high density lipoprotein (HDL) fraction in the plasma irrespective of blood sampling times and administration procedure of Ax. 3. Male broiler chicks (2 weeks of age) were fed on a diet containing 0, 50 or 100 mg/kg of yeast Ax for 2 weeks. Of the tissues examined, Ax concentration in the small intestine was highest, followed by subcutaneous fat, abdominal fat, spleen, liver, heart, kidney and skin. The lowest concentration was in the muscles. Ax concentration in the small intestine, subcutaneous fat, abdominal fat, liver and skin rose as dietary content increased, but this was not the case for the spleen, heart, kidney and muscles except for M. pecloralis superficialis. 4. Over 50% of Ax deposited in liver tissues was detected in the microsomal fraction and 15% was in the mitochondrial fraction. In muscles, both fractions of mitochondria and sarcoplasmic reticulum contained Ax. PMID- 15115212 TI - Smoking cessation: two pilot studies exploring the use of continuous response format algorithm to identify subgroups within stage. AB - Stage of change is typically assessed with a series of yes or no questions, a categorical response format staging algorithm. We revised that algorithm by using, instead, the same questions with 7-point agreement scales covering the first three stages with a sample of smokers. Participants were 141 persons enrolled in introductory psychology classes at the University of Louisville during the fall and spring semesters of 1994. Our continuous response format scores showed expected correlations with modified decisional balance scale scores. In addition, the continuous response format algorithm identified substantial differences in intention to quit sometime and in the next 30 days among participants classified as precontemplators by the categorical system. Analyses from an additional small sample (N = 56) demonstrated relationships between the continuous response format scores and Processes of Change as well as Decisional Balance Scale scores within categorical stages. Subgroups of smokers within stages differ on their attitudes toward changing and on the change strategies they are using. These subgroups can be identified with the continuous response format algorithm. PMID- 15115213 TI - Alexithymia, depressive experiences, and dependency in addictive disorders. AB - Alexithymia, depressive feelings, and dependency are interrelated dimensions that are considered potential "risk factors" for addictive disorders. The aim of this study was to investigate the relationships between these dimensions and to define a comprehensive model of addiction in a large sample of addicted subjects, whether affected by an eating disorder or presenting an alcohol- or a drug use related disorder. The participants in this study were gathered from a multicenter collaborative study on addictive behaviors conducted in several psychiatric departments in France, Switzerland, and Belgium between January 1995 and March 1999. The clinical sample was composed of 564 patients (149 anorexics, 84 bulimics, 208 alcoholics, 123 drug addicts) of both genders with a mean age of 27.3 +/- 8 years. A path analysis was conducted on the 564 dependent patients and 518 matched controls using the scores of the Toronto Alexithymia Scale, the Depressive Experiences Questionnaire, and the Interpersonal Dependency Inventory. Statistical analyses showed good adjustment (Goodness of Fit Index = 0.977) between the observable data and the assumed model, thus supporting the hypothesis that a depressive dimension, whether anaclitic or self-critical, can facilitate the development of dependency in vulnerable alexithymic subjects. This result has interesting clinical implications because identifying specific patterns of relationships leading from alexithymia to dependency can provide clues to the development of targeted strategies for at-risk subjects. PMID- 15115214 TI - Survival of human immunodeficiency virus type 1 after rinsing injection syringes with different cleaning solutions. AB - Bleaching of syringes has been advocated to prevent HIV transmission among injection drug users (IDUs). Several reports indicate that IDUs use household products to disinfect syringes instead of bleach. To test their disinfection efficacy, we performed syringe-rinsing simulations with a range of agents used by IDUs trying to disinfect their syringes. No viable HIV-1 was recovered from syringes rinsed with bleach diluted 1:10. Bleach stored at 37 degrees C and rubbing alcohol performed better than water and the other liquids tested, but less well than bleach 1:10. Rinsing syringes with the other liquids was similar to rinsing with water alone. Increasing the rinsing volume did not always increase the effect of rinsing, but the addition of a second rinse consistently increased rinsing efficacy. Bleaching remains the most effective disinfectant among those tested. It is important that IDUs learn the proper techniques for bleach storage and syringe decontamination. Other household products are not effective disinfectants and should be avoided. Because access to sterile syringes may be restricted by laws, public policy, and police practices, bleach retains its importance in the control of the HIV-1 epidemic among IDUs. PMID- 15115216 TI - Treatment counselor's attitudes about lesbian, gay, bisexual, and transgendered clients: urban vs. rural settings. AB - Treatment counselors' attitudes about lesbian, gay, bisexual, and transgendered (LGBT) clients can have important effects on these client's recovery. There is a common, but unexamined, perception that LGBT people are more accepted in urban areas (and thus urban treatment programs) and that urban counselors have greater knowledge of the needs of the LGBT community. This study examined the attitudes and knowledge of treatment counselors from two geographic regions: urban Chicago (n = 109) and rural Iowa (n = 242) in 2000. The instrument assessed demographic characteristics, knowledge, and experiences working with LGBT clients, and attitudes about LGBT clients (an adaptation of Herek's Attititudes about Lesbians and Gays rating scale). Only a few demographic differences between the urban and rural counselors were identified. Chicago counselors were more racially diverse and more likely to have grown up in an urban area than the Iowa counselors. The Iowa counselors had slightly higher levels of formal education. Although the Chicago providers reported having considerably more contact with LGBT clients and more formal and continuing education about LGBT people, they did not have more positive attitudes or report more knowledge of specific LGBT issues that might influence alcohol and drug treatment. Overall, both Chicago and Iowa counselors had very little formal education regarding the needs of LGBT clients, and nearly half reported negative or ambivalent attitudes. Many of the counselors lacked knowledge about legal issues such as domestic partnership and power of attorney, the concepts of domestic partnership and internalized homophobia, and issues related to family of origin and current family. PMID- 15115215 TI - Validity of the SMAST in two American Indian tribal populations. AB - The standardized evaluation of alcoholism and other psychopathologies in minority populations, particularly American Indians, has long been questioned. This study investigated the validity of one of the most commonly applied assessments for alcoholism--the Short Michigan Alcohol Screening Test (SMAST)--in two distinct American Indian tribal groups. We analyzed data collected from 1989 to 1995 from largely community representative samples of 456 Southwestern and 214 Plains Indians ages 21 or older. For comparison, alcohol dependence was diagnosed using Diagnostic and Statistical Manual of Mental Disorders, Third Edition-Revised (DSM III-R) criteria from a detailed, modified version of the Schedule for Affective Disorders and Schizophrenia--Lifetime (SADS-L). Accuracy of the SMAST was quantified as sensitivity, specificity, likelihood ratios, and the area under the curve for receiver operating characteristics, using the DSM-III-R diagnosis as the reference. The standard SMAST cutoff score of > or = 3 had a demonstrated sensitivity 86% to 95%, but did not perform well in terms of specificity (23% 47%). Significantly higher cutoff scores (> or = 5 for both genders in the Southwestern tribe and 8 and > or = 6 for men and women in the Plains tribe) were required to demonstrate acceptable levels of specificity in both tribes. The findings suggest that the SMAST is not a valid tool to screen for alcohol misuse in these two tribal populations. The highly elevated and different thresholds required from one population to the next and from one gender to the next constitute a significant obstacle to the use of the instrument. PMID- 15115217 TI - You, me...and drugs--a love triangle: important considerations when both members of a couple are abusing substances. AB - Concordant couples can be defined as couples in which both partners have issues with "substance abuse." Studies demonstrate that couples display similar patterns of "substance abuse" that cannot be explained by sociodemographic factors alone. Unfortunately, few studies have focused on the unique relationship dynamics and needs of concordant couples. "Substance abuse" by a client's partner can profoundly affect their recovery and treatment. It is therefore important to understand how clients are influenced by their partners' use. This article attempts to define the needs and issues of concordant couples within a broader psychosocial context. In addition, an overview on the concordant couple literature is provided, along with a discussion of effective treatment and potential barriers to treatment. PMID- 15115218 TI - Understanding exudate management and the role of exudate in the healing process. AB - Studies suggest that wound fluid from acute wounds may have a beneficial effect on wound healing, whereas that of chronic wounds may inhibit healing. Changes in the volume and nature of exudate provide information on the underlying state of the wound and may give an indication of an increasing bacterial load and the presence of infection, and if a wound is likely to proceed to healing. Careful monitoring of the exudate can provide information for the application of systemic and local therapies. Individual wound care products have specific functions which relate to the volume, viscosity and nature of the exudate and these should guide skin care and dressing selection. PMID- 15115219 TI - TIELLE* hydropolymer dressings: wound responsive technology. AB - Wound healing may be compromised by many different factors. Certain underlying illnesses, for example diabetes and malignancy, and wound-specific factors such as critical colonization, infection and protease imbalance can all delay or impair healing. Moist wound healing is now generally accepted in clinical practice to facilitate optimum healing under the correct conditions of wound moisture. Johnson & Johnson Wound Management have provided a solution for the clinician by developing TIELLE* hydropolymer dressings, which provide wound responsive technology for patient care. This unique mode of action achieves positive outcomes for the patient and clinician in terms of cost-effectiveness, patient compliance, healing rates and reduced rates of undesirable adverse effects such as allergic skin reactions, wound pain and maceration. PMID- 15115221 TI - Four case studies of the action of TIELLE* Plus dressings. PMID- 15115220 TI - Clinical evaluation of TIELLE* Plus dressing in the management of exuding chronic wounds. AB - This clinical evaluation, which was carried out between May 2000 and January 2001, measured the efficacy and safety of TIELLE* Plus dressing, a novel hydropolymer dressing with super-absorbent properties. In total 2121 patients with leg ulcers (59.1%), pressure ulcers (20.7%), diabetic foot ulcers (10.9%) or other chronic wounds (9.4%) were included in the study. These were recruited by 624 physicians in a German post-marketing study. All wounds had been present for at least 4 weeks prior to treatment with TIELLE* Plus dressings and many had been treated with other dressings previously. Within the observation period of 12 weeks, 43% of the wounds healed and 50.4% were considered as 'improved'. With this combined total of about 95% the subjects clearly benefited from a change in therapy from conventional regimes (in Germany) such as ointments and gauze, but also from modern, moist wound healing dressings such as hydrocolloids. The frequency of side-effects was low at 4.8%. Over 90% of the patients rated the TIELLE* Plus dressing therapy as 'much better' or 'better' tolerated than the previous treatment regime. For the large majority of the patients the quality of life also improved. On the basis of the positive experiences with respect to effectiveness, safety and handling, 96.8% (604) of the participating doctors wanted to adopt TIELLE* Plus dressings in their therapy plan. TIELLE* Plus dressings can be considered as an effective, safe and simple-to-handle wound dressing for therapy of chronic wounds in daily practice. The shortening of healing time and the less frequent change of dressing also make this therapy regime attractive on cost grounds. PMID- 15115222 TI - Redefining obesity in Asians: more definitive action is required from the WHO. PMID- 15115223 TI - Information technology and medical education in India. PMID- 15115224 TI - Computer use among medical students in an institution in southern India. AB - BACKGROUND: This cross-sectional, descriptive study was conducted in Pondicherry from May to October 2002 to assess computer and internet use among medical students. METHODS: The participants were four batches of undergraduate students, one batch of interns and two batches of postgraduate students. A pre-tested questionnaire was used to collect data from 394 subjects. Written informed consent was obtained from all the participants. RESULTS: Computer knowledge was found to be higher among postgraduates (93.3%) compared to undergraduates (84.5%). Students learnt computers by self-learning, attending classes and using manuals. Writing letters was the most common use of the computer (postgraduates [100%] and undergraduates [87.5%]). Seventy-one per cent of the postgraduates and 43% of the undergraduates used English language dictionaries. About 61% of the undergraduates used computers for playing games. Students also used computer for watching movies. MS Office was the most commonly used software (postgraduates [100%] and undergraduates [72.2%]). The use of software for visual designing and drawing was low. Undergraduates used audio players more often than postgraduates. Most students used the internet for e-mail (postgraduates [100%] and undergraduates [97.3%]). Undergraduate students used the internet for chatting and entertainment. Online banking was also used to some extent. Most of the students were willing to undergo training. All the groups unanimously felt that the institute should provide free training in medical informatics. CONCLUSION: This study revealed high computer use among medical students in an institution with good computer facilities. The majority expressed their willingness to undergo further training. PMID- 15115225 TI - New drugs in India over the past 15 years: analysis of trends. AB - BACKGROUND: New drugs are appearing in the Indian pharmaceutical market every day. To study the trends we analysed the pattern of new drug approvals and introductions in India over the past 15 years (1988-2002). METHODS: Lists of new drugs approved by the Drugs Controller General of India, released half-yearly, were obtained and entered into a computer database. Additional information, such as anatomical therapeutic chemical coding, availability status till 31 December 2002 and source were added to this database before analysing overall time trends and the situation in individual therapeutic categories. RESULTS: Excluding unrecognized and compound formulations and 28 veterinary products, 396 drugs were approved for clinical use during this period. Of these, 315 have also been launched in the market and 5 were subsequently withdrawn. Nervous system-related drugs accounted for the largest number of approvals (82), followed by antimicrobials (73) and cardiovascular drugs (57). Five new antimalarials have emerged but other tropical diseases have been mostly ignored. Eleven vaccines have been added. CONCLUSION: There has been a sharp spurt in the annual number of approvals and introductions. The proliferation of brands and fixed-dose combinations has kept pace with the introduction of new molecules. Unfortunately, most new drugs are not major therapeutic advances. In the context of this rapid proliferation, meeting the information needs of prescribers, establishing an effective nationwide pharmacovigilance system and reorienting the focus of pharmacology education--from information provision to development of self learning and critical judgement skills-are some issues for concern. PMID- 15115226 TI - Experience with telepathology at a tertiary cancer centre and a rural cancer hospital. AB - BACKGROUND: Telepathology allows quick and timely access to an expert opinion, no matter where the patient is located. We analysed the experience with the use of telepathology between a tertiary cancer centre and a rural cancer hospital. METHODS: Ninety-three cases were analysed in which static telepathology was used to obtain a consultation between Tata Memorial Centre and Nargis Dutt Memorial Cancer Hospital at Barshi, a rural area. RESULTS: Successful teleconsultation was achieved in all cases. A diagnosis was offered in 92 cases (98.9%) and was deferred in 1 case (1.1%). Complete concordance, clinically unimportant minor discrepancy and hedged diagnosis were obtained in 83 cases (90.2%). Major discrepancies were encountered in 9 cases (9.7%). The number of images per case ranged from 3 to 27 (average: 7 images). Images were of diagnosable quality in 89.2% of cases. Most of the cases (77.4%) were reported within 3 days; 32.2% were reported within 8 hours (a single working day) and 45.1% within 1-3 days. CONCLUSION: Telemedicine can be effectively used to bridge the gap between medically underprivileged, geographically distant rural areas and advanced centres using the static store and forward methodology. PMID- 15115227 TI - Prevalence of markers of transfusion transmissible diseases in voluntary and replacement blood donors. AB - BACKGROUND: Transfusion of safe blood requires a safe donor. The voluntary donor movement encompasses the concept of a donor who is free from transfusion transmissible infections. It is now mandatory to screen blood for hepatitis B surface antigen, antibodies to HIV-1 and HIV-2, antibodies to hepatitis C virus, syphilis and malarial parasites. METHODS: Between 1996 and 2002, 235 461 donors were screened for markers of hepatitis B virus, and HIV-1 and HIV-2 using commercially available ELISA kits, VDRL test for syphilis and Geimsa stain for the malarial parasite, respectively. A total of 56 476 donors were screened for hepatitis C virus antibodies from June 2001 to December 2002, using third generation ELISA kits. RESULTS: The proportion of voluntary donors increased from 47% to 56% during the study period. The prevalence of HIV showed a steady increase from 0.16% in 1996 to 0.3% in 2002. The prevalence of hepatitis B surface antigen decreased from 1.55% to 0.99%. VDRL reactivity did not show any trend and ranged between 0.11% and 0.66%. Hepatitis C virus antibodies showed a prevalence of 0.4%. The prevalence of all markers was significantly less in voluntary donors. Among the voluntary donors, transfusion transmissible disease markers were significantly less in student donors as compared to other donors. CONCLUSION: A change-over to a voluntary donor service would considerably reduce the number of infectious donors and, among voluntary donors, student donors are the safest. PMID- 15115230 TI - Withdrawal of ventilatory support in the intensive care unit. PMID- 15115228 TI - Phyto-oestrogens and prostatic growth. AB - The incidence of and mortality from prostatic cancer in the West is higher than that in Asian countries. Migrants from Asia to western countries, who maintain their traditional diet, do not have an increased risk of prostatic cancer. This has been attributed in part to 'phyto-oestrogens' in vegetarian Asian diets. Prostatic cancer is a hormone-dependent disease and oestrogens retard the growth of prostatic tumours by interfering with the action of testosterone. Oestrogen increases the level of sex hormone-binding globulin that binds testosterone, resulting in lower free testosterone levels, thereby decreasing androgenic stimulation of the prostate. Oestrogens used to retard the growth of prostatic cancer are associated with certain undesirable side-effects. Phyto-oestrogens have weak oestrogenic potency and anticancer effects. Thus, these phytochemicals have a possible role in the prevention of hormone-dependent diseases such as prostatic cancer. Although the relative potencies of various phyto-oestrogens compared with oestradiol are low, the oestrogen receptor (ER) complexes formed byoestradiol and isoflavones have functional similarities. Also, phyto-oestrogens have a higher affinity to bind to ER-beta than ER-alpha. They are antiproliferative and inhibit tyrosine and other protein kinases which play a key role in tumorigenesis, and also inhibit the production of the potent androgen 5alpha dihydrotestosterone in the prostate. Since prostatic cancer cells usually multiply slowly and the development of this cancer can take many years before symptoms appear, the latent period provides a chemopreventive opportunity for natural therapy with phyto-oestrogens. Although phyto-oestrogens have not yet been used in long-term trials to evaluate their ability to reduce the risk of prostate carcinoma, the evidence thus far suggests that they have a protective effect against the growth of prostate tumours. PMID- 15115229 TI - A new marker of the metabolic syndrome is closer to the heart. PMID- 15115231 TI - And so to books, for new doctors. PMID- 15115232 TI - Emergency medical care: its ethical and legal aspects. PMID- 15115233 TI - Small group discussion followed by presentation as a revision exercise at the end of a teaching module in biochemistry. AB - BACKGROUND: We evaluated the effect of student-dominated small group discussion followed by faculty-moderated presentation as a revision exercise after completion of a teaching module in biochemistry. We assessed the understanding of graduate medical students on the topic and the gain in retention of information, if any, after 15 days. METHODS: Small group discussions involving 11-12 students in each group were conducted on 12 application-oriented problems in 'amino acid metabolism'. A group leader among the students helped to conduct the discussions. While two-thirds of the problems were taken up after the discussion during faculty- or student-moderated interactive presentations, the remaining were not. The effects on low-, medium- and high achievers were evaluated by a pre-test and post-test with multiple choice questions immediately after the session. A subjective feedback was also obtained. To test short-term memory, a post-test with the same multiple choice questions was conducted after 15 days. RESULTS: The exercise was effective and equally beneficial for low-, medium- and high achievers. The gain was maximum when faculty moderated the presentation session. The students' retention of their gain after 15 days was complete. All the students wanted more such sessions in future. CONCLUSION: Student-dominated small group discussion followed by a faculty-moderated presentation is an effective, revision exercise for undergraduate medical students. PMID- 15115234 TI - Healthcare in the Andamans. PMID- 15115235 TI - A tale of two polls. PMID- 15115236 TI - Antimicrobial susceptibility patterns of staphylococci in a tertiary care hospital. PMID- 15115237 TI - Why do doctors make errors? PMID- 15115238 TI - Systemic lupus erythematosus and refractory cytopenia: successful outcome with danazol. PMID- 15115239 TI - Doctored admissions. PMID- 15115240 TI - Bone mineral density in postmenopausal Indian women with Colles fracture. PMID- 15115241 TI - Depression. PMID- 15115242 TI - Maharashtra gets Rs 100 crore to battle HIV. PMID- 15115243 TI - Hyperoxaemia does not change concentrations of serotonin and beta-thromboglobulin in blood of healthy humans. AB - BACKGROUND: The mechanisms of oxygen-induced effects on blood vessels (vasoconstriction in hyperoxaemia and vasodilatation during hypoxaemia) are uncertain. Many investigators have suggested that the vasoconstriction seen during hyperoxia/hyperoxaemia is mediated through the endothelium as a result of either increased release or activity of vasoconstrictors (oxygen radicals, endothelin, norepinephrine, angiotensin II, or serotonin (5-HT)), or reduced activity of vasodilators (prostaglandin E2 and nitric oxide). Serotonin has been assumed to have a central role. METHODS: Eight healthy volunteers were exposed to FiO2 of 1.0 for 20 min and serum concentrations of serotonin and activated platelets were measured (indicated by concentrations of beta-thromboglobulin (beta-TG)). RESULTS: During hyperoxaemia in humans, serum concentrations of scrotonin and beta-TG remained unchanged. CONCLUSION: If serotonin is involved in oxygen-induced vasoconstriction, the mechanism is more likely to be either a potentiating effect of serotonin on other vasoconstrictors or increased activity of serotonin on its receptor. PMID- 15115244 TI - A gel electrophoresis method for detection of mitochondrial DNA mutation (3243 tRNA(Leu (UUR))) applied to a Norwegian family with diabetes mellitus and hearing loss. AB - Blood cells of selected patients from a large Norwegian family with maternally transmitted diabetes mellitus, hearing loss and muscular dysfunction were screened for possible A3243G mutation tRNA(Leu (UUR)) in mitochondrial DNA. We selected 7 patients from 3 of the 4 generations of the family and 10 unrelated healthy control subjects for mutation analysis using denaturing gradient gel electrophoresis (DGGE) and both manual and automated DNA sequencing. The A3243G mutation was found in peripheral blood cells of all 7 patients, but in none of the controls. The mutation was in the form of heteroplasmy and the amount of mutant DNA was found to be between 10% and 35% of total mtDNA in individual patients. This is the first report of a Norwegian family with maternally inherited diabetes and hearing loss carrying the A3243G mutation in mitochondrial DNA. PMID- 15115245 TI - Quantification of prostate specific antigen mRNA levels in circulation after prostatic surgery and endocrine treatment by quantitative reverse transcription polymerase chain reaction. AB - Various methods to detect prostatic cells in circulation have given conflicting results. This is probably because qualitative rather than quantitative methods have been used to detect mRNA from prostatic cells. A quantitative method has been developed based on reverse transcription-polymerase chain reaction (RT-PCR) for detection of prostate specific antigen (PSA) mRNA in peripheral blood. A competitive internal mRNA standard was used for quantification of absolute amounts of PSA mRNA. The detection limit of the assay was 7 copies of mRNA, and the highest level of circulating PSA mRNA in 88 control subjects was 25 copies per milliliter of blood. This method was used to study the influence of prostatic surgery and endocrine treatment on prostatic cells in the circulation of 56 patients undergoing biopsy, radical prostatectomy, transurethral resection of the prostate (TURP), orchiectomy, or androgen blockade. Blood samples were drawn before, during and up to 26 weeks after these procedures had been carried out. The highest level of PSA mRNA in controls was 25 copies per milliliter of blood. After RP, TURP or orchiectomy, PSA mRNA levels increased above this level in 27%, 29%, and 25% of the samples, respectively. After prostate biopsy, two out of 15 patients became positive. PSA mRNA levels that were elevated by surgery became undetectable within 1-3 days. No significant correlation was found between PCR positivity and the clinical characteristics of the patients. It is concluded that the level of PSA mRNA in peripheral blood increases after prostatic surgery, indicating temporary dissemination of prostatic cells. However, preoperative levels do not correlate with serum PSA, stage or grade. PMID- 15115246 TI - Extracellular matrix protein expression in cerebrospinal fluid from patients with tropical spastic paraparesis associated with HTLV-I and Creutzfeldt-Jakob disease. AB - The cerebrospinal fluid (CSF) is in direct contact with the extracellular space of the CNS, thus biochemical processes in the CNS could potentially be reflected in the CSF. Changes in extracellular matrix (ECM) proteins can be studied through their analysis in the CSF. ECM plays an essential role in CNS homeostasis and several proteins such as laminin (LN), fibronectin (FN), thrombospondin (TS) and heparan sulphate proteoglycan (HS, perlecan) form part of its structure. Possible changes in the levels of these proteins were investigated in two different pathologies--tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM) (n=25) and Creutzfeldt-Jakob disease (CJD) (n=19)--and compared with those in a control group with or without neurological disease (n=25). CSF analyses were carried out using monoclonal or monospecific polyclonal antibodies. In comparison with the control group, it was found that TSP/HAM patients presented significantly higher levels of LN, TS and HS, while in CJD patients the levels of FN, TS and HS were increased. In CJD patients the HS level was almost double that of the TSP/HAM patients. These results suggest a distinct pattern of ECM proteins in CSF in relation to the type of neurological disease. TSP/HAM is a chronic motor disease that affects the white matter of the spinal cord, while CJD is a subacute dementia that affects cerebral neurons and their synapsis. PMID- 15115247 TI - The Microfiltrometer (MicroFM): a new filtration device for the assessment of less deformable erythrocyte subpopulations. AB - The hardware of a new filtration device, the Microfiltrometer (MicroFM), is described. The different components of the device; impedance meter, power supply, measuring cell and its 5-micron Oligopore filter are described and it is shown how they are interrelated and interfaced to a computer for data acquisition. The properties of the filter and the general functioning principle of the device are also elucidated. For each run, the MicroFM generates elementary signals from individual passages of many hundreds of red blood cells (RBCs) through micropores of a given 5-micron Oligopore filter. Analysis of each elementary signal provides two complementary parameters, the transit time tau of the explored RBC and the change in electrical impedance deltaZ caused by the temporary flow of the considered RBC through a particular micropore of the filter. These two parameters can be utilized for reliable assessment of erythrocyte deformability on a cellular level. PMID- 15115248 TI - Salivary cortisol and administration of concentrated oral glucose in newborn infants: improved detection limit and smaller sample volumes without glucose interference. AB - Newborn infants are subject to repetitive painful and stressful events during neonatal intensive care. When the baby attempts to cope with a stressful situation the hypothalamus-pituitary-adrenal axis is activated, releasing cortisol. The free cortisol response is optimally measured in saliva and saliva samples can be taken easily and without pain. However, saliva is very scarce in infants and saliva stimulants can interfere with analytical methods. Nowadays, sweet solutions are frequently administered to neonates prior to a disturbing procedure in order to reduce pain. The possible interference of sweet solutions with the measurement of salivary cortisol has not yet been documented. The aims of the present study were to further improve the detection limit of the radioimmunoassay used for cortisol analysis and to determine the degree of interference of high concentrations of glucose with the analytical method. By decreasing incubation temperature and prolonging the incubation time it was possible to improve the detection limit of the radio immunoassay (RIA) to 0.5 nmol/L at the same time as the sample volume was decreased to 10 microL saliva. Saliva was collected from full-term and preterm babies and was sufficient for analysis in 113 out of 116 (97%) samples. Glucose in the concentrations and amounts commonly used for pain relief did not interfere with the RIA method. In conclusion, it is feasible to collect microlitre volumes of saliva and analyse even very low concentrations of cortisol in newborns. It is also possible to offer the baby oral glucose prior to a painful procedure and still reliably measure salivary cortisol. PMID- 15115249 TI - No effects of acute hyperglycaemia and hyperinsulinaemia on skin microcirculation and endothelial markers in Type II diabetes mellitus. AB - BACKGROUND: Increased microvascular permeability is a hallmark of microangiopathy in Type I diabetes mellitus and is associated with endothelial dysfunction and haemodynamic alterations. Type II diabetes mellitus is characterized by insulin resistance and hyperinsulinaemia. The purpose of this study was to determine whether acute hyperinsulinaemia, under both normoglycaemic and hyperglycaemic conditions, increases skin capillary permeability through its effect on skin haemodynamics, capillary recruitment or circulating markers of endothelial dysfunction in Type II diabetes. METHODS: Nine Type II diabetic patients without microalbuminuria, (pre-) proliferative retinopathy or clinical neuropathy underwent three glucose clamps of 210 min., in random order, on separate days. A "standard" clamp (insulin-infusion rate 30 mU kg(-1) h(-1), glucose-target 5.0 mmol/L) was compared with a hyperinsulinaemic (insulin-infusion rate 150 mU kg( 1) h(-1), glucose-target 5.0 mmol/L) and a hyperinsulinaemic, hyperglycaemic (insulin-infusion rate 150 mU kg(-1) h(-1), glucose-target 12.0 mmol/L) clamp. Skin capillary permeability and density were measured using large-window sodium fluorescein videodensitometry, and skin blood flow by laser Doppler flowmetry. Endothelial dysfunction was estimated from increases in soluble intercellular adhesion molecule-1 (sICAM-1) and von Willebrand factor antigen (vWF). RESULTS: No differences were found in skin capillary permeability, skin haemodynamics and capillary density at the end of the three glucose clamp periods. sICAM-1 and vWF did not increase as compared to the standard glucose clamp. sICAM-1 (r=-0.76, p<0.05) and vWF (r=-0.71, p<0.05) correlated negatively with insulin sensitivity, but not with skin microcirculatory parameters. CONCLUSIONS: Acute hyperinsulinaemia, both with and without concomitant hyperglycaemia, does not increase skin microvascular permeability, haemodynamics or parameters of endothelial dysfunction in Type II diabetic patients. Furthermore, these data suggest that the coexistence of hyperinsulinaemia and endothelial dysfunction in Type II diabetes does not indicate a causal relationship, but may rather indicate decreased insulin sensitivity as a common underlying cause. PMID- 15115250 TI - Evaluation of the blood analyzer ABL 735 radiometer for determination of the percentage of fetal hemoglobin in fetal and neonatal blood. AB - OBJECTIVE: To evaluate the quality of measurements of percentage fetal hemoglobin (HbF) using the blood analyzer ABL 735 and to compare the results with the HbF measurements using the Biorad-Variant (the gold standard). METHODS: The percentage of HbF in fetal blood, obtained during intrauterine transfusions or from the umbilical cord blood of preterm and term babies, and from the red blood cells of adults who had given blood for transfusion, was determined spectrophotometrically (ABL-735) and by high pressure liquid chromatography (Biorad-Variant). Blood pH values were also measured with the ABL 735. Reproducibility (mean difference between duplo measurements) and the relationships between the HbF percentages measured by both methods were calculated. RESULTS: Reproducibility was independent of the HbF percentage and was more reliable with the Biorad-Variant method (0.3%) than with the ABL-735 method (2.1%). Regression analysis showed that the ABL-HbF percentages were often underestimated. Serious aberrations were observed at low pH values (<7.00) on the recalculated ABL-HbF percentages. For samples with pHs >7.00 a relatively good relationship was found between the pH-corrected ABL-HbF percentages and those of the Biorad-Variant values. CONCLUSION: The HbF percentage measured with the ABL blood analyzer is only moderately accurate if the HbF percentage is corrected for the pH. PMID- 15115251 TI - Non-physiological levels of circulating cortisol in growth hormone-treated hypopituitary adults after conventional cortisone substitution. AB - OBJECTIVE: To assess the usefulness of measuring plasma cortisol profiles in growth hormone-treated hypopituitary adults and to compare these with cortisol levels in healthy controls. METHODS: Eleven ACTH-deficient adult patients received 12.5 mg cortisone-acetate orally at 16.00 h and 25 mg at 07.00 h. The patients arrived in the ward at 12.00 h. After tablet intake at 16.00 h, samples for serum cortisol were taken at hourly intervals for the next 24 h, except between 07.00 and 12.00 h when samples were drawn every half hour; 24-h urinary free cortisol (24-h-UFC) excretion was collected simultaneously. For comparison, 8 healthy controls were investigated. RESULTS: The patients had circulating cortisol levels with very low plasma cortisol at 07.00 h before their morning dose of cortisone acetate. At the same time period, controls had their highest plasma cortisol levels. After tablet intake the patients had a rapid initial absorption of cortisol, but a marked variability in the morning peak levels (Cmax), and the Cmax was in general higher and occurred 90 min later than the Cmax in the controls. The 24-h-UFC excretion and 24-h area under the curve (24-h AUC) did not differ between patients and controls. The female patients had higher 24-h-AUC for plasma cortisol (p=0.032) and tended to have higher plasma cortisol peaks in the morning, but had levels of 24-h-UFC similar to those of the male patients. CONCLUSIONS: Conventional cortisone substitution with a twice-daily replacement regimen in hypopituitary adults results in abnormal circulating cortisol profiles with low or non-measurable morning values and variable individual peaks. This suggests that the present dosing schemes have to be improved and that cortisone substitution should be individualized. PMID- 15115252 TI - Comparison of a peanut agglutinin test and an immunochemical faecal occult blood test in detecting colorectal neoplasia in symptomatic patients. AB - BACKGROUND: Currently available methods for detection of early-stage colorectal cancer are reliant on faecal occult blood (FOB) tests. Bleeding, however, is not specific for colorectal neoplasia. Enzymatically detected or peanut agglutinin (PNA)-detectable galactose-beta1-3-N-acetyl-galactosamine residues found in rectal mucus have been used to detect colorectal cancer. METHODS: The sensitivity and specificity of the PNA rectal mucus test were compared with those of an immunological test for faecal occult blood (Hemolex) in 199 symptomatic patients referred for colorectal investigations. All patients also underwent a colonoscopy. SDS-PAGE and PNA-overlay were used to characterize PNA-binding proteins in normal and malignant colorectal tissue. RESULTS: The PNA test had a similar sensitivity to that of Hemolex for colorectal carcinoma (83% vs. 72%), adenomas (55% vs. 50%), inflammatory bowel disease (52% vs. 48%) and hyperplastic polyps (48% vs. 25%). The sensitivity of the PNA test and Hemolex for colorectal neoplasia was 69% vs. 59% and specificity 68% vs. 86% (p=0.002). SDS-PAGE and PNA overlay showed some commonly expressed PNA-binding proteins in both normal mucosa and colorectal cancer and a higher and even selective expression of 160 kD PNA binding protein in colorectal cancer. CONCLUSIONS: A single PNA test in its present form is as sensitive an indicator of colorectal neoplasia as Hemolex completed over three days, but lacks specificity. The 160 kD cancer-associated antigen we have identified is under further characterization for development of a more specific PNA test. PMID- 15115253 TI - Immunoassays of human trefoil factors 1 and 2: measured on serum from patients with inflammatory bowel disease. AB - BACKGROUND: The trefoil factors (TFF1-3) are cysteine-rich peptides expressed in the gastrointestinal tract where they play a critical role in mucosal protection and repair. The expression is up-regulated at sites of ulceration in various chronic inflammatory diseases. Recently, we presented an ELISA method for measurement of TFF3. The aims of the present study were to develop and evaluate ELISAs for the other two known human trefoil peptides, TFF1 and TFF2, and to carry out a cross-sectional study on serum TFF levels in patients with inflammatory bowel disease (IBD). METHODS: The TFF1-ELISA was based on two polyclonal rabbit antibodies and the TFF2-ELISA on a monoclonal mouse antibody and a polyclonal rabbit antibody. RhTFF1 and 2 were employed to prepare the calibrators. TFF1-3 were assayed in serum from IBD patients (n=41) and controls (n=13). RESULTS: The TFF1- (TFF2-) ELISA had a detection limit of 3 pmol/L (6 pmol/L) and an analytical imprecision (CV(A)) of 7.0-8.8 for mean concentrations of 24-120 pmol/L (6.1-8.0 for mean concentrations of 17-77 pmol/L). The central reference intervals (n=300) were 140-1400 pmol/L (37-190 pmol/L). There was no variation with age and menstrual cycle. Food intake reduced concentrations of TFF1 by approximately 15%, but did not influence concentrations of TFF2. TFF1 and TFF3 were increased in serum from IBD patients. CONCLUSIONS: We have developed assays for measuring TFF1 and TFF2. Finding increased TFF concentrations in serum from IBD patients suggests that measurements of trefoil peptides may be of clinical relevance in IBD. PMID- 15115254 TI - Gc globulin (vitamin D-binding protein) levels: an inhibition ELISA assay for determination of the total concentration of Gc globulin in plasma and serum. AB - Gc globulin, also called vitamin D-binding protein, is a plasma protein involved in the actin-scavenger system. In this study, the total Gc globulin concentration in serum or plasma samples was determined using a new, fast, solid-phase inhibition assay. Included in the study were 228 healthy volunteers (131 M, 97 F), 22 pregnant women, 90 cancer patients and 9 patients with chronic liver disease. Moreover, the degree of complexing with actin was determined in selected samples using crossed immunoelectrophoresis. The Gc globulin level in healthy controls was in the range 176-623 mg/L, showing no age dependency. The median level was found to be significantly higher in women than in men. Gc globulin concentrations were raised during pregnancy, showing a median value of 541 mg/L in the first trimester, and slightly raised to 574 mg/L in the second trimester. Cancer patients showed no changes in Gc globulin level, and there was no sign of increased amounts of complexing with actin. Chronic liver patients showed increased levels of Gc globulin following transplantation, but no signs of complexing with actin. This new solid-phase inhibition assay is fast, it is a good complement to the existing quantification methods, and it is especially suitable for determination of the Gc globulin status in acute liver patients before and during treatment. PMID- 15115255 TI - Idiopathic left ventricular tachycardia with a right bundle branch block morphology and left axis deviation ("Belhassen type"): results of radiofrequency ablation in 18 patients. AB - BACKGROUND: Idiopathic left ventricular tachycardia with a right bundle branch block configuration and left axis deviation, first described by Belhassen et al., is a rare electrocardiographic-electrophysiologic entity. Radiofrequency catheter ablation has been proposed as a good therapeutic option, but the best criteria for determining the optimal site of ablation are still under debate. OBJECTIVES: To report the clinical features, electrophysiologic characteristics, results of RFA, and long-term outcome in 18 patients with "Belhassen's VT" treated in our laboratory during the last 10 years, stressing the best electrophysiologic criteria for determining the optimal site of ablation. METHODS: Eighteen consecutive patients with this specific VT underwent RFA in our laboratory during the last 10 years. RFA was acutely successful in 17 patients after one or two procedures (15 and 2 patients, respectively) using 4.1 +/- 2.2 RF pulses. The putative ablation sites were defined by good pace-mapping (3 patients), earliest recorded Purkinje spike prior to the QRS onset during VT or sinus rhythm (6 patients), earliest endocardial activation during VT (1 patient), and diastolic potential preceding the Purkinje spike during VT and/or late diastolic potential in sinus rhythm (7 patients). In the patients with a definite successful ablation, the ratio of successful to unsuccessful radiofrequency pulse delivery to the diastolic potential site was compared to that of other methods. The ratio of successful RFA at the diastolic potential site (5:8) was higher than in the other methods (8:31) and the difference was statistically significant (P = 0.05). Successful ablation sites were more basal when the diastolic potential site was chosen. CONCLUSION: The results of the present study confirm the high success rate and safety of RFA using conventional techniques in the management of "Belhassen VT," suggesting that this procedure can be used as a first-line therapy. Ablating at a site demonstrating a late diastolic potential is at least as effective as ablating at a ventricular exit site, although the use of combined electrophysiologic criteria may be the optimal approach. PMID- 15115256 TI - Clinical manifestations and outcome of Pseudomembranous colitis in an elderly population in Israel. AB - BACKGROUND: Pseudomembranous colitis is a well-recognized cause of diarrhea in patients receiving antibiotics and has significant consequences in terms of morbidity, mortality and cost. Clostridium difficile infection is the single most important infectious cause of PMC. PMC is frequently nosocomial, with an increased risk of spread among institutionalized patients, both in hospitals and nursing homes. OBJECTIVE: To investigate the demographic, clinical and laboratory characteristics of PMC patients in an Israeli elderly population. METHODS: We studied 72 hospitalized patients with endoscopically proven PMC. The medical records of all patients including clinical history and laboratory data were reviewed, such as: age, pre-hospitalization status (dependency or not, in the community as compared to the nursing home), background medical history, presenting symptoms, antibiotic history, physical examination on admission, hematologic and biochemical parameters, treatment, duration of hospitalization, complications, mortality, and recurrence of disease. RESULTS: Of the 72 patients (34 males and 38 females, mean age 77 years) 47% were nursing home residents. Pre hospitalization antibiotic treatment was given to 91.4% for infections of the upper respiratory tract (45%) and urinary tract (45%). The most common antibiotics were cephalosporin (64%), penicillins (42%) and quinolones (28%). Sixty-four percent of the patients were treated with more than one antibiotic, 26% of patients received anti-acid therapy and 36% had been fed with a nasogastric tube. On admission, leukocytosis was found in 79% of patients, > 20,000/mm3 in half of them; 60% were anemic, 60% had elevated erythrocyte sedimentation rate, and 78% had hypoalbuminemia. Treatment consisted of metronidazole (41%) or a combination of metronidazole and vancomycin (56%). Overall, 31% of patients recovered without complications, 29% died within 30 days of hospitalization, and 24% were re-hospitalized due to recurrence of PMC. CONCLUSION: The most common antibiotics implicated in PMC are cephalosporin, penicillins and quinolones. The disease is associated with high mortality and recurrence rates. PMID- 15115257 TI - Visceral leishmaniasis in Israel, 1960-2000. AB - BACKGROUND: Visceral leishmaniasis was first reported in Israel (then Palestine) in 1929. In the 1960s and 1970s, it was endemic to northern Israel, but only partial data about the disease have been gathered since then. OBJECTIVE: To investigate the epidemiologic trends of visceral leishmaniasis in Israel from 1960 to 2000, and to delineate some clinical features of the infection. METHODS: Data were collected from hospital charts, scientific publications, and reports of the Ministry of Health and the Kuvin Center for the Study of Infectious and Tropical Diseases. RESULTS: During the last four decades, 87 cases of visceral leishmaniasis were diagnosed in Israel, 76 of them (87%) in children. In the 1960s, all 54 patients were diagnosed in the northern part of the country. The rate of infection declined significantly in the 1970s (5 cases) and then increased slightly in the 1980s (11 cases) and 1990s (17 cases). More than 50% of the cases in the 1990s were in central Israel. Children accounted for 100% of cases in the 1960s but only 58% in the 1990s. The main clinical features of the patients diagnosed in the last decade were fever, weight loss, hepatosplenomegaly and pancytopenia. Three of the adults were co-infected with human immunodeficiency virus. DISCUSSION: The decline in the incidence of visceral leishmaniasis in the 1970s and the slight increase in the 1980s and 1990s can be attributed to changes in the animal reservoir and vectors, and in the immunity status of part of the population exposed to Leishmania. CONCLUSIONS: Visceral leishmaniasis has reemerged in Israel. This mandates better control of the animal reservoir and vectors and increased awareness to this infection. PMID- 15115258 TI - Atopic dermatits in infants and children in Israel: clinical presentation, allergies and outcome. AB - BACKGROUND: Atopic dermatitis is a common disease in infants and children and the incidence appears to be rising. OBJECTIVES: To determine the presentation, allergies, and outcome among Israeli infants and children. METHODS: Children with atopic dermatitis referred to the allergy clinic at a regional pediatric center were evaluated for their medical history and their allergy. The allergic assessment was determined by utilizing skin prick tests and/or serum specific immunoglobulin E concentrations. The children were reexamined again for all parameters at the end of the follow-up period. RESULTS: Forty-six children with atopic dermatitis were studied, 27 males (58.7%) and 19 females (41.3%). A family history of allergy was found in 19 (41.3%). The median age at presentation was 17 months. Of the 46 children 33 (71.7%) revealed an allergy to one or more of the allergens. The most common combination was allergy to food and house-dust mites. The mean follow-up time was 64 months. By the age of 8 years full recovery was seen in 16 patients, half of whom recovered within 3.3 years from the date of presentation. The probability of complete remission was 58%, and for either complete or partial remission 76%. Upon reevaluation at the end of the follow-up period some patients lost their sensitivities, while others, who had been allergic to foods, became sensitive to house-dust mites and/or pollens. CONCLUSIONS: Atopic dermatitis is an allergic problem in the northem region of Israel, as it is in other parts of the world. Food allergy and house-dust mites are major contributors to the evolution of eczema. PMID- 15115259 TI - Comparative study of response to treatment with supraphysiologic doses of B vitamins in hyperhomocysteinemic hemodialysis patients. AB - BACKGROUND: Hyperhomocysteinemia is a well-recognized risk factor for accelerated atherosclerosis in hemodialysis patients. OBJECTIVES: To examine the effects of two doses of vitamins B6 and B12 and folic acid on homocysteine levels in hemodialysis patients and assess the functional impact of the methylenetetrahydrofolate reductase genotype on the response to treatment. METHODS: In a randomized prospective study, we assessed the effects of folic acid and two doses of B-vitamins in 50 hemodialysis patients with hyperhomocysteinemia. Patients were divided into two groups: 26 patients (group A) who received 25 mg of vitamin B6 daily and one monthly injection of 200 microg vitamin B12, and 24 patients (group B) who received 100 mg of vitamin B6 daily and one monthly injection of 1,000 microg vitamin B12. In addition, both groups received 15 mg folic acid daily. Patients were evaluated for homocysteine levels as well as for coagulation and a thorough lipid profile. Baseline Hcy levels were determined after at least 4 weeks washout from all folic acid and B-vitamins that were given. MFTHR alleles were analyzed, as were activated protein C resistance, von Willebrand factor and lupus anticoagulant. RESULTS: Basal plasma Hcy levels were significantly elevated in hemodialysis patients compared with normal subjects (33.8 +/- 4.3 vs. 4.5 to 14.0 micromol/L). Following treatment, Hcy levels were significantly reduced to 21.2 +/- 1.6 in group A and 18.6 +/- 1.4 micromol/L in group B (P < 0.01). There was no difference in Hcy reduction following the administration of either high or low dosage of vitamins B6 and B12 utilized in the present study. There was no correlation between Hcy levels or thrombophilia and high incidence of thrombotic episodes in hemodialysis patients. Genotypic evaluation of MTHFR revealed that the presence of homozygous thermolabile MTHFR (n = 5) was associated with higher Hcy levels and better response to treatment (Hcy levels decreased by 58%, from 46.2 +/- 14.6 to 19.48 + 4.1 micromol/ L following treatment). In patients with heterozygous thermolabile MTHFR (n = 25), Hcy levels decreased by 34%, from 31.2 +/- 3.7 to 18.1 +/- 1.1 micromol/L following treatment. The efficacy of high and low doses of B-vitamins on the reduction of homocysteine levels was comparable. CONCLUSIONS: Treatment with B-vitamins in combination with folic acid significantly decreased homocysteine levels in hemodialysis patients, independently of the tested doses. In addition, mutations in MTHFR were associated with elevated plasma levels of Hcy. Neither vascular access nor the presence of diabetes was associated with higher pre- or post-treatment homocysteine level. PMID- 15115260 TI - Systemic lupus erythematosus and thyroid disease. PMID- 15115261 TI - Perinuclear antineutrophil cytoplasmic autoantibodies and anti-Saccharomyces cerevisiae antibodies: serologic markers in inflammatory bowel disease. PMID- 15115262 TI - Efficacy of omega-3 fatty acid supplementation in primary and secondary prevention of coronary heart disease. PMID- 15115263 TI - A reentry story. PMID- 15115264 TI - Omega-3: the vanishing nutrient beyond cardiovascular prevention and treatment. AB - Omega-3 appears to be an important nutrient component of the mammalian body, however because of changes in the food chain during the last century it has become increasingly rare in frequently eaten foods. Currently, the main source of omega-3 is fish, which tends to be expensive and is periodically found to be contaminated. Common foods such as eggs, chicken, etc., which were once a rich source of omega-3, are now lacking it. Given the importance of omega-3 in a variety of body functions, as well as in the prevention of disease, it is obvious that in the coming years the scientific community worldwide will have to target agricultural research and development to the enrichment of foods with omega-3. PMID- 15115265 TI - Can the laryngeal mask airway replace endotracheal intubation for airway control? The argument for the laryngeal mask airway. PMID- 15115266 TI - Can the laryngeal mask airway replace endotracheal tubation for airway control? The argument against the laryngeal mask airway. PMID- 15115267 TI - Single lung transplantation in refractory asthma with irreversible airflow obstruction. PMID- 15115268 TI - Infant botulism in Israel: knowledge enables prompt diagnosis. PMID- 15115269 TI - Endoscopically assisted wireless capsule endoscopy in a patient with familial adenomatous polyposis after total proctocolectomy and continent ileostomy. PMID- 15115270 TI - Transient bilateral cortical blindness in an adult after an acute episode of asthma exacerbation. PMID- 15115271 TI - Pulmonary hypertension and sleep apnea. PMID- 15115272 TI - Complementary and alternative medicine. PMID- 15115273 TI - Complementary and alternative medicine. PMID- 15115274 TI - Litigation as a quality improvement tool. PMID- 15115275 TI - The business case for quality: combining medical literature research with health plan data to establish value for nonclinical managers. AB - Clinical managers face a growing need to communicate the value of what they do in terms that can be interpreted by nonclinical financial managers. We have sought to link the evidence basis of current guidelines to variables that will demonstrate in more financial terms the very real benefit of treating diseases aggressively. We have developed an approach using the medical literature that is designed to describe clinical initiatives in more concrete terms as desired by senior management. This becomes specifically critical during budget time and when justification for various clinical programs is needed. The approach uses medical research from the peer-reviewed literature to estimate the economic impact of various initiatives and then combines the analysis with an organization's actual data to impute potential benefit. A sample grid for developing the analysis is attached. A comprehensive bibliography that will assist others with similar endeavors has been included. Although not as rigorous as formal methods, actuarial analyses, or health services research activities, it presents a beginning framework around which an organization can create operational estimates of initiative effectiveness. PMID- 15115276 TI - A review of physician turnover: rates, causes, and consequences. AB - To assess current knowledge about physician turnover, this article reviews the available literature regarding the rates, causes, and consequences of physicians' leaving a practice. Reported rates of turnover vary widely, both because of differences in reporting methods and by type of healthcare organization and provider. A common contributor to turnover is a mismatch between physicians' expectations and organizational culture or rules, although greater understanding of ways to assess and to predict such mismatch is needed. The costs and consequences of physician turnover are substantial and may include: (a) financial consequences, (b) effects on patient satisfaction, and (c) effects on the organization or practice as a whole, related to the experience of other healthcare providers, and effects on institutional public relations. Our review suggests that greater attention to physician turnover is needed to clarify its frequency and predictors, and strategies to lessen its occurrence. PMID- 15115278 TI - Factors associated with the successful implementation of a quality improvement project in human immunodeficiency virus ambulatory care clinics. AB - We examined a quality improvement (QI) program, offered to ambulatory care clinics (N = 82) serving human immunodeficiency virus-positive clients, to determine what factors predicted the clinic independently implementing QI processes without their program consultant's help. Initial analyses examined clinics at 4 levels of involvement: withdrew from the project, initial QI proficiency, advanced QI proficiency, and consultant independent. The initial and advanced stages were collapsed into 1 group (consultant dependent) and compared with consultant-independent clinics for multivariate logistic regression. In the multivariate models, 3 factors significantly predicted the clinic being consultant independent: staffing level (odds ratio [OR] = 1.6, 95% confidence interval [CI] = 1.2-2.2), the number of participating months (OR = 1.4, 95% CI = 1.0-2.0), and baseline QI readiness (OR = 1.1, 95% CI = 1.0-1.3). Receiver operator curves were calculated for significant predictors; the strongest predictor was staffing (c statistic = .79). Clinics that are organizationally prepared for QI, allow adequate time to adopt QI methods into their organization, and provide adequate QI staffing are more likely to independently apply QI methods. PMID- 15115277 TI - Patient and staff safety: voluntary reporting. AB - Central to efforts to assure the quality of patient care in hospitals is having accurate data about quality and patient problems. The purpose was to describe the reporting rates of medication administration errors (MAE), patient falls, and occupational injuries. A questionnaire was distributed to staff nurses (N = 1105 respondents) in a national sample of 25 hospitals. This addressed voluntary reporting, work environment factors, and reasons for not reporting occurrences. More than 80% indicated that all MAEs should be reported, but only 36% indicated that near misses should be reported. Perceived levels of actual reporting were: 47% of MAEs, 77% of patient falls, 48% of needlesticks, 22% of other exposures to body fluids, and 17% of back injuries. Administrative response to reports, personal fears, and unit quality management were related to reporting. Patient and staff safety occurrences are underreported. Strong quality management processes and positive responses to reports of occurrences may increase reporting and enhance safety. PMID- 15115279 TI - Quality improvement in family practice: a recipe for change. AB - A new recipe is presented, splitting quality improvement into 4 levels. The Q1 level corresponds to the everyday processes that guide our daily work flow. Q2 corresponds to commonly thought of outcome measures such as HEDIS criteria. Q3 relates to the executive functions that permit seasoned clinicians to draw generalizations about care for individual patients by synthesizing large amounts of data from both psychosocial as well as classical history/physical sources. Finally Q4 reflects more population-based quality improvement activities. Examples are given for each. Each of these levels requires a different approach for improvement activities. Each must be seen in the context of an expanded "quality compass" and in the paradigm of the PLAN-DO-CHECK-ACT cycle of quality improvement. Finally, a practical application of how this could be instituted at a Family Practice residency is given. PMID- 15115280 TI - Introducing Professor Masataka Mori, Asia-Australian Regional Editor. PMID- 15115281 TI - Overexpression of heat shock proteins differentially modulates protein kinase C expression in rat neonatal cardiomyocytes. AB - Previous studies have suggested that protein kinase C (PKC) is involved in heat shock protein (Hsp)-mediated cardioprotection. Therefore, we wanted to determine whether overexpression of Hsps modulates PKC expression, which will give us further insight into understanding the mechanism by which Hsps and PKC interact to protect cells from stress-induced injury. Specifically, we overexpressed the inducible form of Hsp70 (Hsp70i) or Hsp90 in rat neonatal cardiomyocytes and evaluated PKCdelta or PKCepsilon expression by immunoblotting and immunofluorescent confocal microscopy. Western analysis showed that overexpression of Hsp70i or Hsp90 decreased PKCepsilon expression. However, overexpression of Hsp70i or Hsp90 did not modify PKCdelta expression over control levels. Overexpression of constitutively active PKCdelta or PKCepsilon increased Hsp70i expression over control levels. The data suggest that overexpression of Hsps differentially modulates expression of PKC isoforms in rat neonatal cardiomyocytes. Furthermore, PKC may directly play a role in Hsp-mediated cardioprotection by upregulating Hsp70i expression. PMID- 15115282 TI - CHIP: a link between the chaperone and proteasome systems. AB - CHIP, carboxy terminus of Hsc70 interacting protein, is a cytoplasmic protein whose amino acid sequence is highly conserved across species. It is most highly expressed in cardiac and skeletal muscle and brain. The primary amino acid sequence is characterized by 3 domains, a tetratricopeptide repeat (TPR) domain at its amino terminus, a U-box domain at its carboxy terminus, and an intervening charged domain. CHIP interacts with the molecular chaperones Hsc70-Hsp70 and Hsp90 through its TPR domain, whereas its U-box domain contains its E3 ubiquitin ligase activity. Its interaction with these molecular chaperones results in client substrate ubiquitylation and degradation by the proteasome. Thus, CHIP acts to tilt the folding-refolding machinery toward the degradative pathway, and it serves as a link between the two. Because protein degradation is required for healthy cellular function, CHIP's ability to degrade proteins that are the signature of disease, eg, ErbB2 in breast and ovarian cancers, could prove to be a point of therapeutic intervention. PMID- 15115283 TI - Mechanisms for regulation of Hsp70 function by Hsp40. AB - The Hsp70 family members play an essential role in cellular protein metabolism by acting as polypeptide-binding and release factors that interact with nonnative regions of proteins at different stages of their life cycles. Hsp40 cochaperone proteins regulate complex formation between Hsp70 and client proteins. Herein, literature is reviewed that describes the mechanisms by which Hsp40 proteins interact with Hsp70 to specify its cellular functions. PMID- 15115284 TI - Molecular characterization of genes encoding cytosolic Hsp70s in the zygomycete fungus Rhizopus nigricans. AB - Previous studies have shown that some stressors, including steroid hormones 21-OH progesterone and testosterone, stimulate the accumulation of heat shock protein 70 (hsp70) messenger ribonucleic acid (mRNA) population in the zygomycete filamentous fungus Rhizopus nigricans. In this study we report the cloning of 3 R nigricans hsp70 genes (Rnhsp70-1, Rnhsp70-2, and Rnhsp70-3) encoding cytosolic Hsp70s. With a Southern blot experiment under high stringency conditions we did not detect any additional highly homologous copies of the cytosolic hsp70 genes in the R nigricans genome. Sequence analyses showed that all 3 genes contain introns within the open reading frame. The dynamics of the R nigricans molecular response to progesterone, 21-OH progesterone, and testosterone, as well as to heat shock, copper ions, hydrogen peroxide, and ethanol was studied by temporal analysis of Rnhsp70-1 and Rnhsp70-2 mRNA accumulation. Northern blot experiments revealed that the Rnhsp70-2 transcript level is not affected by testosterone, whereas mRNA levels of both genes are rapidly increased with all the other stressors studied. Moreover, the decrease of transcript levels is notably delayed in ethanol stress, and a difference is observed between the profiles of Rnhsp70-1 and Rnhsp70-2 transcripts during heat stress. PMID- 15115285 TI - Expression of heat shock protein 32 (hemoxygenase-1) in the normal and inflamed human stomach and colon: an immunohistochemical study. AB - Heat shock protein 32 (Hsp32, hemoxygenase-1) is induced by reactive oxygen metabolites (ROM) and degrades heme leading to the formation of antioxidant bilirubin. Increased mucosal generation of ROM occurs in gastritis and inflammatory bowel disease. We aimed to assess mucosal expression of Hsp32 in normal stomach and colon and to test the hypothesis that disease-related differential expression occurs in inflamed tissue. Gastric body and antral mucosal biopsies were obtained from 33 patients comprising Helicobacter pylori negative normal controls (n = 8), H pylori-negative gastritis patients (n = 11), and H pylori-positive gastritis patients (n = 14). Forty-seven archival colonic mucosal biopsies selected comprised normal histology (n = 10), active ulcerative colitis (UC) (n = 9), inactive UC (n = 8), active Crohn's disease (CD) (n = 8), inactive CD (n = 6), and other colitides (n = 6). Hsp32 expression in formalin fixed sections was assessed by avidin-biotin peroxidase immunohistochemistry using a polyclonal rabbit anti-Hsp32 as the primary antibody. Immunohistochemical staining identified Hsp32 in all groups. Diffuse cytoplasmic staining was seen in gastric and colonic epithelial and lamina proprial inflammatory cells. Staining scores for Hsp32 were higher in antral H pylori-positive (P = 0.002) and H pylori negative (P = 0.02) gastritis than in controls and in body H pylori-positive gastritis than in the other 2 groups (P < 0.01). Expression of Hsp32 was increased in active UC compared with inactive disease (P = 0.03) and normal controls (P = 0.02). In conclusion, Hsp32 is expressed constitutively in normal gastric and colonic mucosa, and differential expression occurs in these tissues when they are inflamed. Upregulation of Hsp32 may be an adaptive response to protect mucosa from oxidative injury in patients with gastritis and inflammatory bowel disease. PMID- 15115286 TI - Heat shock protein 70 or heat shock protein 27 overexpressed in human endothelial cells during posthypoxic reoxygenation can protect from delayed apoptosis. AB - Overexpression of heat shock protein (Hsp) 70 and Hsp27 in vivo was proclaimed as a potential tool in therapy of ischemia-reperfusion injury. However, it was so far not known whether these Hsps can beneficially act when increased in cells just at the stage of postischemic reperfusion. This issue was examined in a model of ischemia-reperfusion stress when cultures of endothelial cells (EC) from human umbilical vein were infected with virus-based vectors expressing Hsp70 or Hsp27, or Hsp56, or green fluorescent protein (GFP) and exposed to 20 hours of hypoxia followed by reoxygenation. The infection was performed either 10 hours before hypoxia or immediately after hypoxia, or at different time points of reoxygenation. Only low cell death was detected during hypoxia, but later, up to 40% of the treated cells died via caspase-dependent apoptosis between 6 and 12 hours of reoxygenation. The percentage of apoptotic cells was 1.6- to 3-fold greater in Hsp56- and GFP-infected EC than in Hsp70- or Hsp27-infected EC. The last 2 groups exhibited a lesser extent of procaspase-9 and procaspase-3 activation within 6-9 hours of reoxygenation. The cytoprotective effects of overexpressed Hsp70 and Hsp27 were observed not only in the case of infection before hypoxia but also when EC were infected at the start of reoxygenation or 1 2 hours later. An increase in the Hsp70 and Hsp27 levels in infected EC correlated well with their resistance to apoptosis under reoxygenation. These findings suggest that overexpression of Hsp70 or Hsp27, if it occurs in the involved cells at the early stage of postischemic reperfusion, can still be cytoprotective. PMID- 15115287 TI - Heat shock protein 70-reactivity is associated with increased cell surface density of CD94/CD56 on primary natural killer cells. AB - Previously we described an involvement of the C-type lectin receptor CD94 and the neuronal adhesion molecule CD56 in the interaction of natural killer (NK) cells with Hsp70-protein and Hsp70-peptide TKD. Therefore, differences in the cell surface density of these NK cell-specific markers were investigated comparatively in CD94-sorted, primary NK cells and in established NK cell lines NK-92, NKL, and YT after TKD stimulation. Initially, all NK cell types were positive for CD94; the CD56 expression varied. After stimulation with TKD, the mean fluorescence intensity (mfi) of CD94 and CD56 was upregulated selectively in primary NK cells but not in NK cell lines. Other cell surface markers including natural cytotoxicity receptors remained unaffected in all cell types. CD3-enriched T cells neither expressing CD94 nor CD56 served as a negative control. High receptor densities of CD94/CD56 were associated with an increased cytolytic response against Hsp70 membrane-positive tumor target cells. The major histocompatibility complex (MHC) class I-negative, Hsp70-positive target cell line K562 was efficiently lysed by primary NK cells and to a lower extent by NK lines NK-92 and NKL. YT and CD3-positive T cells were unable to kill K562 cells. MHC class-I and Hsp70-positive, Cx + tumor target cells were efficiently lysed only by CD94-sorted, TKD-stimulated NK cells with high CD94/CD56 mfi values. Hsp70-specificity was demonstrated by antibody blocking assays, comparative phenotyping of the tumor target cells, and by correlating the amount of membrane bound Hsp70 with the sensitivity to lysis. Remarkably, a 14-mer peptide (LKD), exhibiting only 1 amino acid exchange at position 1 (T to L), neither stimulated Hsp70-reactivity nor resulted in an upregulated CD94 expression on primary NK cells. Taken together our findings indicate that an MHC class I-independent, Hsp70 reactivity could be associated with elevated cell surface densities of CD94 and CD56 after TKD stimulation. PMID- 15115288 TI - Deoxyribonucleic acid damage induced by doxorubicin in peripheral blood mononuclear cells: possible roles for the stress response and the deoxyribonucleic acid repair process. AB - Doxorubicin is an antineoplastic drug widely used in cancer treatment. However, many tumors are intrinsically resistant to the drug or show drug resistance after an initial period of response. Among the different molecules implicated with doxorubicin resistance are the heat shock proteins (Hsps). At present we do not know with certainty the mechanism(s) involved in such resistance. In the present study, to advance our knowledge on the relationship between Hsps and drug resistance, we have used peripheral blood mononuclear cells obtained from healthy nonsmoker donors to evaluate the capacity of a preliminary heat shock to elicit the Hsp response and to establish the protection against the deoxyribonucleic acid (DNA) damage induced by doxorubicin. DNA damage and repair were determined using the alkaline comet assay. We also measured the expression of Hsp27, Hsp60, Hsp70, Hsp90, hMLH1, hMSH2, and proliferating cell nuclear antigen by immunocytochemistry. The damage induced by doxorubicin was more efficiently repaired when the cells were previously heat shocked followed by a resting period of 24 hours before drug exposure, as shown by (1) the increased number of undamaged cells (P < 0.05), (2) the increased DNA repair capacity (P < 0.05), and (3) the high expression of the mismatch repair (MMR) proteins hMLH1 and hMSH2 (P < 0.05). In addition, in the mentioned group of cells, we confirmed by Western blot high expression levels of Hsp27 and Hsp70. We also noted a nuclear translocation of Hsp27 and mainly of Hsp70. Furthermore, inducible Hsp70 was more expressed in the nucleus than Hsc70, showing a possible participation of Hsp70 in the DNA repair process mediated by the MMR system. PMID- 15115289 TI - Progression of arteriovenous bypass restenosis in mice exposed to a 50 Hz magnetic field. AB - The controversy over whether magnetic fields (MF) produced by electrical wiring and appliances contribute to diseases such as cancer has been debated in the literature for more than 2 decades. These extremely low frequency fields at 50 or 60 Hz are omnipresent in the industrialized world and have been linked to various forms of cancer by epidemiological studies. Little has been published investigating any possible role of MF and cardiovascular disease, and this is the first study looking specifically at the effect of exposure to high-intensity MF on the development and progression of restenosis. A mouse arteriovenous bypass model was used, and mice were exposed to MF for periods of 1, 2, or 3 weeks. Neointima formation, infiltration of mononuclear cells, and heat shock protein 60 expression were all studied at the conclusion of the exposure regimen. Animals exposed to the MF for 1 week showed significantly smaller neointima formation compared with control mice exposed to a null field, although this difference was not observed in mice exposed for 2 or 3 weeks. No difference was found between mice exposed to MF and controls in any of the other parameters investigated. PMID- 15115290 TI - Tomato heat stress protein Hsp16.1-CIII represents a member of a new class of nucleocytoplasmic small heat stress proteins in plants. AB - We describe a new class of plant small heat stress proteins (sHsps) with dominant nuclear localization (Hsp17-CIII). The corresponding proteins in tomato, Arabidopsis, and rice are encoded by unique genes containing a short intron in the beta4-encoding region of the alpha-crystallin domain (ACD). The strong nuclear localization results from a cluster of basic amino acid residues in the loop between beta5 and beta6 of the ACD. Using yeast 2-hybrid tests, analyses of native complexes of the sHsps, and immunofluorescence data, we demonstrate that, in contrast to earlier observations (Kirschner et al 2000), proteins of the sHsp classes CI, CII, and CIII interact with each other, thereby influencing oligomerization state and intracellular localization. PMID- 15115291 TI - Spatial analysis of cell death and Hsp70 induction in brain, thymus, and bone marrow of the hyperthermic rat. AB - Heat shock response and programmed cell death are cellular reactions to stressful stimuli. Previous studies have not correlated these responses in vivo at the spatial level in mammalian tissues. This study uses a dual procedure involving immunocytochemistry for Hsp70 localization and the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end-labeling (TUNEL) assay for cell death to correlate the pattern of stress-inducible Hsp70 and cell death at the cellular level. After whole-body hyperthermia in the rat, an increase in Hsp70-positive cells and TUNEL-positive cells was noted in brain, thymus, and bone marrow. However, 2 populations of cells were apparent in the tissues examined, those inducing Hsp70 and those triggered into programmed cell death. Cells that were both Hsp70 positive and TUNEL positive were rarely detected. In tissues of the intact mammal, cells that induce Hsp70 after whole-body hyperthermia were not triggered into programmed cell death. PMID- 15115292 TI - UV radiation effects on pathogens and insect pests of greenhouse-grown crops. PMID- 15115293 TI - UV radiation effects on pathogens and insect pests of greenhouse-grown crops. AB - Production of high-value crops is often performed under protected cultivation. In recent years various spectral modifications have been made in greenhouse covers. Two of the main reasons to modify the spectral characteristics of greenhouse covers have been to suppress the proliferation of several foliar diseases and to protect crops from insects and insect-borne virus diseases of greenhouse-grown crops. These goals were achieved by complete or partial absorption of solar UV radiation, which interrupts the life cycle of several fungal pathogens and alters the visual behavior of many insects. Examples of these management strategies are described in this article. PMID- 15115294 TI - Binding, aggregation and photochemical properties of methylene blue in mitochondrial suspensions. AB - Methylene Blue (MB) has well-established photochemical properties and has been used in a variety of photochemical applications including photodynamic therapy. Despite the fact that most of MB's cytotoxic effects in cells are attributed to mitochondrial damage, the interactions of this dye with mitochondria and the consequent effects on photochemical properties have not yet been fully determined. We monitored MB binding, aggregation and its ability to release singlet oxygen (1O2) on irradiation when interacting with mitochondrial suspensions. MB actively binds to mitochondria and enters the matrix in a manner stimulated by the mitochondrial proton potential and by the increase in mitochondrial concentrations. The greater accumulation of MB in mitochondria with elevated proton potentials or those treated with high concentrations of MB results in the formation of MB dimers, previously shown to be less effective generators of 1O2. Accumulation of MB within mitochondria with high membrane potentials also results in the reduction of MB to the photochemically inactive leuco-MB. Indeed, irradiation of mitochondria with high proton potentials in the presence of MB results in the generation of approximately half the quantity of 1O2 compared with 1O2 generated in mitochondria with low proton potentials. These differences in photochemical properties should influence the cytotoxic effects of photodynamic treatment in the presence of MB. PMID- 15115295 TI - Effect of pH on inhibition and enhancement of luminol-H2O2-Co2+ chemiluminescence by phenolic compounds and amino acids. AB - The effect of pH on inhibition and enhancement of luminol-H2O2-Co2+ chemiluminescence (CL) by 18 phenolic compounds and 20 amino acids was studied. It was found that most of the tested compounds showed an inhibiting effect at lower pH and an enhancing effect at higher pH. At a midrange pH, for some phenolic compounds with two ortho-position -OH, both an inhibiting and an enhancing peak were simultaneously observed. UV-visible spectra of the tested phenolic compounds at different pH values were studied. The mechanism for CL inhibition and enhancement was proposed. It is likely that the competition of the -OH or the -NH2 group and other reducing groups in the molecules with luminol for O2*- led to the CL inhibition. A reaction of -COO(-) and quinone or ketone formed by phenolic compounds at higher pH via deprotonation with O2*- also resulted in the CL enhancement. PMID- 15115296 TI - pH-dependent modification of lipophilicity of porphyrin-type photosensitizers. AB - Structural modifications of photosensitizers (changes in protonation, ionic state and aggregation state) under different environmental conditions should be precisely determined to understand the interaction of the photosensitizers with biological systems. In the present study partition coefficients of hematoporphyrin IX (HpIX), disulfonated meso-tetraphenylporphine, meso-tetra(3 hydroxyphenyl)porphine (mTHPP) and meso-tetra(3-hydroxyphenyl)chlorin in the 1 octanol-phosphate buffer system were determined in the pH region 4.0-8.0. Only the partition coefficients of HpIX and mTHPP were found to be pH dependent. Computer processing of fluorimetric titration data was applied to estimate pKa values of the imino nitrogens of mTHPP. Monoprotonated species of mTHPP seem to be unstable or nonexistent. The possibility that both imino nitrogens of this dye are protonated according to a common pKa is proposed. The pKa value of the imino nitrogens of mTHPP was found to be 2.99 +/- 0.04 after the application of a model taking aggregation of the drug into account. The contributions of various aqueous ionic species of mTHPP as functions of pH were calculated and compared with partition coefficients. PMID- 15115297 TI - Photophysical and photobiological behavior of antimalarial drugs in aqueous solutions. AB - This article describes the results of a combined photophysical and photobiological study aimed at understanding the phototoxicity mechanism of the antimalarial drugs quinine (Q), quinacrine (QC) and mefloquine (MQ). Photophysical experiments were carried out in aqueous solutions by stationary and time-resolved fluorimetry and by laser flash photolysis to obtain information on the various decay pathways of the excited states of the drugs and on transient species formed on irradiation. The results obtained showed that fluorescence and intersystem crossing account for all the adsorbed quanta for Q and MQ (quantum yield of about 0.1 and 0.9, respectively) and only for 24% in the case of QC, which has a negligible fluorescence quantum yield (0.001). Laser flash photolysis experiments evidenced, for QC and MQ, the occurrence of photoionization processes leading to the formation of the radical cations of the drugs. The effects of tryptophan and histidine on the excited states and transient species of the three drugs were also investigated. In parallel, the photoactivity of the antimalarial drugs was investigated under UV irradiation on various biological targets through a series of in vitro assays in the presence and in the absence of oxygen. Phototoxicity on 3T3 cultured fibroblasts and lipid photoperoxidation were observed for all the drugs. The photodamage produced by the drugs was also evaluated on proteins by measuring the photosensitized cross-linking of spectrin. The combined approaches were proven to be useful for understanding the mechanism of phototoxicity induced by the antimalarial drugs. PMID- 15115298 TI - Preliminary study on the photoproduction of hydroxyl radicals in aqueous solution with Aldrich humic acid, algae and Fe(III) under high-pressure mercury lamp irradiation. AB - Under a high-pressure mercury lamp (HPML) and using an exposure time of 4 h, the photoproduction of hydroxyl radicals (*OH) could be induced in an aqueous solution containing humic acid (HA). Hydroxyl radicals were determined by high performance liquid chromatography using benzene as a probe. The results showed that *OH photoproduction increased from 1.80 to 2.74 microM by increasing the HA concentration from 10 to 40 mg L(-1) at an exposure time of 4 h (pH 6.5). Hydroxyl radical photoproduction in aqueous solutions of HA containing algae was greater than that in the aqueous solutions of HA without algae. The photoproduction of *OH in the HA solution with Fe(III) was greater than that of the solution without Fe(III) at pH ranging from 4.0 to 8.0. The photoproduction of *OH in HA solution with algae with or without Fe(III) under a 250 W HPML was greater than that under a 125 W HPML. The photoproduction of *OH in irradiated samples was influenced by the pH. The results showed that HPML exposure for 4 h in the 4-8 pH range led to the highest *OH photoproduction at pH 4.0. PMID- 15115299 TI - Changes in matrix gene and protein expressions after single or repeated exposure to one minimal erythemal dose of solar-simulated radiation in human skin in vivo. AB - Damage to the skin extracellular matrix (ECM) is the hallmark of long-term exposure to solar UV radiation. The aim of our study was to investigate the changes induced in unexposed human skin in vivo after single or repeated (five times a week for 6 weeks) exposure to 1 minimal erythemal dose (MED) of UV solar simulated radiation. Morphological and biochemical analyses were used to evaluate the structural ECM components and the balance between the degrading enzymes and their physiologic inhibitors. A three-fold increase in matrix metalloproteinase 2 messenger RNA (mRNA) (P < 0.02, unexposed versus exposed) was observed after both single and repeated exposures. Fibrillin 1 mRNA level was increased by chronic exposure (P < 0.02) and unaltered by a single MED. On the contrary, a single MED significantly enhanced mRNA levels of interleukin-1alpha (IL-1alpha), IL-1beta (P < 0.02) and plasminogen activator inhibitor-1 (P < 0.05). Immunohistochemistry demonstrated a significant decrease in Type-I procollagen localized just below the dermal-epidermal junction in both types of exposed sites. At the same location, the immunodetected tenascin was significantly enhanced, whereas a slight increase in Type-III procollagen deposits was also observed in chronically exposed areas. Although we were unable to observe any change in elastic fibers in chronically exposed buttock skin, a significant increase in lysozyme and alpha-1 antitrypsin deposits on these fibers was observed. These results demonstrate the existence of a differential regulation, after chronic exposure compared with an acute one, of some ECM components and inflammatory mediators. PMID- 15115300 TI - A far-red fluorescent contrast agent to image epidermal growth factor receptor expression. AB - Recent developments in optical technologies have the potential to improve the speed and accuracy of screening and diagnosis of curable precancerous lesions and early cancer, thereby decreasing the costs of detection and management of epithelial malignancies. The development of molecular-specific contrast agents for markers of early neoplastic transformation could improve the detection and molecular characterization of premalignant lesions. In the oral cavity, epidermal growth factor receptor (EGFR) overexpression has been identified in early stages of premalignant lesions of the oral squamous cell carcinoma; therefore, real-time assessment of EGFR expression could serve as a biomarker for oral neoplasia. The purpose of our study was to develop a molecular-specific optical contrast agent targeted against EGFR for in vivo assessment of epithelial neoplasia using a monoclonal antibody and the far-red fluorescent dye, Alexa Fluor 660 streptavidin. In addition to demonstrating the specificity of the contrast agent for EGFR in cell lines, we document the ability to achieve penetration through 500 microm thick epithelial layers using multilayer tissue constructs and permeability-enhancing agents. Finally, using the fluorescence intensity of the contrast agent on fresh oral cavity tissue sections, we were able to distinguish abnormal from normal oral tissue. This contrast agent should have important clinical applications for use in conjunction with fluorescence spectroscopy or imaging (or both) to facilitate tumor detection and demarcation. PMID- 15115301 TI - The role of carotenoids in the photoadaptation of the brown-colored sulfur bacterium Chlorobium phaeobacteroides. AB - The brown-colored sulfur bacterium Chlorobium (Cb.) phaeobacteroides 1549 (new name, Chlorobaculum limnaeum 1549) contains many kinds of carotenoids as well as bacteriochlorophyll (BChl) e. These carotenoids were identified with C18-high performance liquid chromatography, absorption, mass and proton nuclear magnetic resonance spectroscopies and were divided into two groups: the first is carotenoid with one or two phi-end groups such as isorenieratene and beta isorenieratene and the second is carotenoid with one or two beta-end groups such as p-zeacarotene, beta-carotene and 7,8-dihydro-beta-carotene. The latter 7,8 dihydro-beta-carotene was found to be a novel carotenoid in nature. OH-gamma Carotene glucoside laurate and OH-chlorobactene glucoside laurate were also found as minor components. The distribution of BChl e homologs in Cb. phaeobacteroides cultivated under various light intensities did not change, but the carotenoid to BChl e ratio changed markedly: carotenoid with the phi-end group maintained the same ratio to BChl e, whereas that with the beta-end group increased with increasing light intensity. The cells cultured under low-light intensity contained more phi-end carotenoids than beta-end. In Cb. phaeobacteroides the wavelength of the Qy band of BChl e aggregates did not change. We suggested that Cb. phaeobacteroides photoadapts to light intensity by changing the carotenoid composition. PMID- 15115302 TI - Interexperimental and interindividual variations of DNA repair capacities after UV-B and UV-C irradiations of human keratinocytes and fibroblasts. AB - DNA repair plays a central role in the cellular response to UV. In this work we have studied the response of skin cells (i.e. fibroblasts and keratinocytes) from the same or from different individuals after both ultraviolet-B (UV-B) and ultraviolet-C (UV-C) irradiations using the comet assay to characterize the specific cellular response to UV-induced DNA damage. Cells were irradiated with increasing doses of UV-B or UV-C. To study the UV dose dependency of initial steps of DNA repair, namely recognition and incision at DNA damage level, the comet assay was performed, under alkaline conditions, 60 min after UV irradiation to allow detection of DNA strand breaks. Comparative analysis of tail moment values after UV exposure of cells from the same or from different individuals showed interexperimental and interindividual variations, implying that repeated assays are necessary to characterize the individual DNA repair capacity. With increasing doses of UV in keratinocytes, a plateau was rapidly reached after irradiation, whereas in fibroblasts a linear dose-effect relationship was observed. These interindividual variations associated with cellular specificity in DNA response may be of significance in skin cell and individual susceptibility toward UV-induced carcinogenesis. PMID- 15115303 TI - Effects of acid and alkali on the light absorption, energy transfer and protein secondary structures of core antenna subunits CP43 and CP47 of photosystem II. AB - The effects of acid and alkali treatment on the light absorption, energy transfer and protein secondary structure of the photosystem II core antenna CP43 and CP47 of spinach were investigated by the absorption spectra, fluorescence emission spectra and circular dichroism spectra. It has been found that acid treatment caused the appearance of absorption characteristic of pheophytin a (Pheo a), whereas alkali treatment induced a new absorption peak at 642 nm. The energy transfer between beta-carotene and chlorophyll a (Chl a) in CP43 was easily disturbed by alkali, whereas in CP47 was readily affected by acid. As to the effects on the secondary structure of proteins in CP43 and CP47, effects of acid were far less than those of alkali. Both acid and alkali disturbed the microenvironment of Chl a and interfered exciton interaction between Chl a molecules. It was suggested that acid and alkali affect the light absorption, energy transfer and protein secondary structure of CP43 and CP47 in a different way. H+ can permeate into the internal space of alpha-helix, change Chl a into Pheo a and disturb the microenvironment of pigments without damaging the secondary structure of protein, whereas OH- can induce the protein unfolding at first, then saponify Chl a to chlorophyllide and disturb the microenvironment of pigments. PMID- 15115304 TI - Effect of monovalent and divalent cations on the photoinactivation of bacteria with meso-substituted cationic porphyrins. AB - It is well established that for successful photoinactivation (PI) of gram negative bacteria a cationic photosensitizer is required. This requirement suggests a charge-dependent interaction between the photosensitizer and the gram negative bacterium, which may be influenced by the presence of ions in the suspending medium. The aim of the present study was to investigate the effect of cations Na+ and Ca2+ on the efficacy of the PI of the gram-negative Pseudomonas aeruginosa and the gram-positive Staphylococcus aureus. The bacteria were suspended in buffer containing either meso-tetra(N-methyl-4-pyridyl)-porphyrin or meso-mono-phenyl-tri(N-methyl-4-pyridyl)-porphyrin as photosensitizer and various concentrations of Na+ or Ca2+. The cell suspensions were exposed to a broadband light dose of 9 J/cm2. In buffer without added cations, P. aeruginosa and S. aureus were equally sensitive to PI. Addition of cations strongly decreased the sensitivity of both bacteria to PI, with the PI of P. aeruginosa being much more decreased than that of S. aureus, and Ca2+ being more effective than Na+. The decreased sensitivity was accompanied by a reduced binding of the photosensitizers to the bacteria. PMID- 15115305 TI - The absence of CD44 ameliorates Fas(lpr/lpr) disease. AB - The lpr mutation in the Fas gene leads to symptoms of autoimmune disease, including polyclonal B-lymphocyte activation and lymphoproliferation. The expanding T-lymphocyte populations in the disease are characterized by high expression levels of the memory marker CD44. It has not been known whether CD44 expression contributes to the pathophysiology of the condition or merely reflects a consequence of excessive lymphocyte activation. We therefore tested the role of CD44 gene products in Fas(lpr/lpr) disease in gene targeted mice. We bred CD44 knockout mice and C57Bl/6-lpr mice to generate the Fas(lpr/lpr) CD44+/+, and Fas(lpr/lpr) CD44-/- genotypes and analyzed the disease manifestations around 215 days of age. The absence of CD44 substantially reduced the immunoglobulin secretion and lymphocyte expansion that are characteristic of the Fas(lpr/lpr) syndrome. Surprisingly, the percentage of CD3+ CD4- CD8- (double negative) cells in peripheral lymphoid organs increased in Fas(lpr/lpr) CD44-/- mice almost to the same extent as in Fas(lpr/lpr) CD44+/+ mice. These results indicate that the expansion of the fraction of double negative cells in spleens and lymph nodes, believed to be generated by down-regulation of CD8, does not depend on increased lymphocyte numbers. Furthermore, they corroborate an essential role for CD44 gene products in the T-cell expansion and polyclonal B-cell activation that constitute the Fas(lpr/lpr) syndrome. The CD44 receptor may be a suitable therapeutic target for inhibition of lymphoproliferation in autoimmune diseases. PMID- 15115306 TI - Common patterns of B cell perturbation and expanded V4-34 immunoglobulin gene usage in autoimmunity and infection. AB - Features of the lymphocyte population in systemic lupus erythematosus (SLE) include a disordered B cell profile and production of autoantibodies. An additional distinctive perturbation is the overexpression of V4-34-encoded serum immunoglobulins (Ig). A similar rise in V4-34-encoded Ig occurs in normal subjects following infection with certain herpesviruses, and is found in Epstein Barr virus (EBV)-associated infectious mononucleosis (IM). To assess common and distinctive features of B cells in patients with SLE and IM, we compared the B cell profile and V4-34 gene involvement in patients with SLE and IM. B cell profiles from patients with IM paralleled those of patients with SLE, showing a differential loss of naive and memory B cells and the maintenance of plasmablast/early plasma cells. Class-switched V4-34-encoded IgG from plasmablast/early plasma cells was evident both in patients with SLE and IM and revealed common features of oligoclonal expansions with most having undergone somatic hypermutation. It has been proposed that, in healthy individuals, expression of the V4-34 gene is specifically censored prior to isotype switch as a control on autoreactivity. If so, censoring is bypassed following EBV infection, after which equilibrium is restored. Continuing high serum levels in SLE may arise either by disordered regulation, or by subclinical reactivation of endogenous virus. PMID- 15115307 TI - Interaction between gonadal steroids and neuroimmune system in acute experimental autoimmune encephalomyelitis (EAE) in Wistar rats. AB - Experimental autoimmune encephalomyelitis (EAE) is an inflammatory disease of the CNS mediated by autoreactive T lymphocytes directed against myelin antigens. Since neuroendocrine-immune dysfunction appears to contribute to the pathogenesis of autoimmune diseases, the present work was designed to study the effect of changes in the endocrine system on the development of acute EAE and the immune response against myelin basic protein (MBP). Intact and sham males and intact female Wistar rats showed the most severe clinical symptoms (acute period) 12-14 days post-inoculation (dpi). Then, they began gradually to recover, regaining the total ability to walk by 15-17 dpi. Male Wistar rats with altered levels of gonadal hormones by surgical castration showed an onset of the symptoms retarded 2-3 days with respect to the other EAE groups, showing neuropathological symptoms up to 27-28 dpi, and remaining with lower body weight even at 40 dpi. The castrated animals exhibited a specific delay in MBP-stimulated DTH reactivity that correlates with the delay in the onset of the clinical symptoms. Also significant lymphocyte proliferation to MBP was still present at 35 dpi that was absent in the sham group. The distribution of the IgG subclasses indicated that at 35 dpi castrated animals have a higher IgG2b/IgG1 ratio (35.1) in comparison to that presented by sham rats (4.8). Considering that at this time the castrated animals were not completely recuperated, these results could indicate an ongoing inflammatory immune response associated with Th1 activity in these animals. Also castrated animals developed antibodies to a diversity of MBP epitopes in comparison to sham rats, which presented a dominance of antibodies to MBP peptide p96-128. These results indicate that sex hormones levels regulate cell-mediated immunity and the specificity of anti-MBP antibodies related to the induction and development of acute EAE. PMID- 15115308 TI - A second heavy chain permits survival of high affinity autoreactive B cells. AB - Anti-DNA antibody is the serological hallmark of systemic lupus erythematosus (SLE). While antibodies with this specificity may be generated in many individuals, only patients with SLE fail to regulate them effectively. We have demonstrated previously that in non-autoimmune mice transgenic for the heavy chain of the R4A-gamma2b anti-DNA antibody, the existence of high affinity, IgG2b dsDNA binding B cells is tightly correlated with the co-expression of endogenous IgM heavy chain. These cells are anergic. In contrast, low affinity IgG2b dsDNA binding B cells do not express an endogenous heavy chain and represent a population of immunocompetent autoreactive B cells. In order to determine whether the presence of a second heavy chain permits the high affinity autoreactive B cells to escape deletion, the R4A-gamma2b mouse was mated to a strain with a targeted deletion of the transmembrane portion of the mu heavy chain, muMT mice, to produce R4A-gamma2b/muKO mice. Serum titers of anti-DNA antibodies were negligible in both R4A-gamma2b and R4A-gamma2b/muKO mice. In R4A-gamma2b/muKO mice, however, LPS was able to activate a DNA-reactive population although an LPS inducible DNA-reactive population. Light chain gene usage in transgene expressing B cells from R4A-gamma2b/muKO mice was similar to that of the previously defined low affinity anti-DNA B cells that escape tolerance. These data suggest a requirement for a second heavy chain for the survival of this anergic B cell subset. PMID- 15115309 TI - Association between insulin resistance and GAD65-autoantibody levels--a pilot study in an adult non-diabetic population. AB - The aim of this pilot study was to investigate any association between insulin resistance (IR) and serum levels of autoantibodies against the glutamic acid decarboxylase (GAD65Ab) among adult non-diabetic subjects. Based on calculations of IR using the IR homeostasis model in a Swedish adult non-diabetic population (n = 756) participating in the WHO MONICA-study, an insulin sensitive group (n = 54, M/F:27/27) and an insulin resistant group (n = 46 M/F:24/22) were identified. Serum from the subjects were analysed for the presence of GAD65Ab. There was no significant difference in GAD65Ab levels between the groups. However, there was a correlation between IR and serum GAD65Ab within the insulin sensitive group (Spearman rho 0.4, p < 0.01). Our observation could indicate that IR could serve as an initiator or a progression factor in the autoimmune process in subjects predisposed to autoimmunity. This finding will be further investigated in a larger study including subjects with a continuum of IR. PMID- 15115310 TI - Flow cytometric analysis of the LE cell phenomenon. AB - A flow cytometry-based phagocytosis assay was developed and utilized to measure the LE cell phenomenon at the single cell level in vitro. Since the lupus erythematosus (LE) cell phenomenon is a special form of necro-phagocytosis in the presence of anti-dsDNA antibodies, dead substrate cells or chicken erythrocytes nuclei (CEN) served as targets that were labeled with propidiumiodide (PI). Phagocytes (PMN) were stained by anti-CD45 mAb FITC. After co-incubation phagocytosis was measured by flow cytometry. Flow cytometric analysis enabled the discrimination between PI+/CD45- targets, PI-/CD45+ phagocytes, and PI+/CD45+ phagocytes with engulfed targets. Maintaining the samples on ice significantly reduced the phagocytic uptake as compared to samples co-cultivated at 37 degrees C (p < 0.0002). The phagocytic up-take was lowest after substrate pre-treatment in normal serum as compared to samples with either no serum exposure or pre treatment in LE-serum with anti-dsDNA antibodies (p < 0.05). Taken together, these data suggest the phagocytosis-based flow cytometry assay is suitable for analyzing the LE cell phenomenon. This method provides an interesting, simple and rapid new tool, and will possibly alleviate further studies on the LE cell phenomenon with modified cell models and/or conditions. PMID- 15115311 TI - Anti-insulin activity in IgG-fractions from children with newly-diagnosed type 1 diabetes and negative for insulin autoantibodies. AB - Insulin autoantibodies (IAA) are often detected as the first humoral sign of beta cell autoimmunity in prospective studies in young children with increased genetic risk of type 1 diabetes. After the appearance of IAA their level typically rise but seems to decline in many cases before the clinical presentation of type 1 diabetes. We hypothesized that the reason for the sudden drops in the levels of IAA could be the formation of immune complexes caused by binding of antibodies to free insulin in plasma. We studied whether isolation of the IgG-fraction and dissociation of immune complexes by acid treatment using protein A column results in the appearance of detectable IAA in those children with newly-diagnosed type 1 diabetes whose plasma samples test negative for IAA. IAA assay was performed in IgG-fractions and corresponding plasma samples from 17 children with type 1 diabetes and 23 unaffected children all testing negative for plasma IAA. The levels of IAA measured from IgG-fractions of diabetic children were higher than the levels of IAA measured from IgG-fractions in the control children (p = 0.004 in Mann-Whitney U-test). Forty-seven percent (8 out of 17) of newly-diagnosed patients negative for plasma IAA before IgG separation had increased levels of IAA in IgG-fractions and only 13% (3 out of 23) of controls. The levels of glutamate decarboxylase autoantibodies (GADA) did not differ between patients (n = 14) and controls (n = 21) negative for plasma GADA when measured in IgG fractions. Our results suggest that formation of immune complexes results in false negative results in tests for IAA but not for GADA. PMID- 15115312 TI - Polymorphisms of the TGF-beta1 promoter in tight skin (TSK) mice. AB - TGF-beta1 plays a major role in fibrotic diseases including scleroderma. Human fibroblasts from sclerotic lesions display an increased sensitivity to TGF-beta1. Similarly, fibroblasts from TSK mice which develop a scleroderma-like syndrome are hyperresponsive to TGF-beta. The aim of the present study was to investigate whether the TGF-beta hypersensitivity demonstrated by TSK/+ fibroblasts is associated with polymorphisms of the TGF-beta1 promoter. Sequence analysis revealed one polymorphism (a G --> T at -1133 bp) unique to the TSK/+ mouse. Transfection of fibroblasts with a 1.8 kb fragment of the TGF-beta1 promoter containing the -1133 polymorphism exhibited increased basal TGF-beta1 promoter activity which was enhanced upon incubation with TGF-beta1. This may be related to the loss of a negative regulatory site in the TSK/+ TGF-beta1 promoter. PMID- 15115313 TI - The L1 retroelement-related p40 protein induces p38delta MAP kinase. AB - We characterized a full length L1 mRNA in a rheumatoid arthritis (RA) synovial tissue and determined the degree of methylation of its 5'-UTR. We asked whether not only intact but also altered L1s can exert biological activities by transfecting RA synovial fibroblasts (SF) with either retrotransposition competent or incompetent L1s and examined their capacity to induce p38delta. Total RNA was isolated from the synovial tissue of a 35-year-old woman with highly destructive RA. A complete L1 sequence was obtained by 3'/5'-RACE. Methylation of the genomic 5'-UTR was determined by the sodium-disulfide/PCR method. RA-SF were transfected by lipofection with either a functional L1 or an ORF2-mutated L1 element. The expression of p38delta was measured by RT-PCR and Western blot. The full length L1 mRNA included a 5'-UTR, an ORF1 and an ORF2. Three of five CpG islands (60%) of the genomic L1 5'-UTR were hypomethylated and the ORF2 was deactivated by the insertion of stop codons. Both, intact and ORF2 mutated L1 vectors, induced the expression of p38delta. Thus, even an ORF2 mutated L1 element, as expressed in RA, is biologically active and both L1 ORF1 and p38delta transcripts may appear as a consequence of genomic hypomethylation. The induction of p38delta appears to be mediated by an ORF1/p40-dependent process. This is the first indication of a p40 mediated transactivation. PMID- 15115314 TI - Paradoxical exacerbation of psoriasis in AIDS: proposed explanations including the potential roles of substance P and gram-negative bacteria. AB - Psoriasis, a TH1-induced disorder, is not more common in human immunodeficiency virus (HIV) infection than in the general population. However, it may appear for the first time or pre-existing psoriasis may worsen and be difficult to treat in HIV disease. The paradoxical exacerbation of psoriasis in AIDS has not been fully explained. Various explanations have been proposed including (a) the reduction of Langerhans' cells (LCs) in HIV disease, (b) the direct epidermal proliferative effect of HIV, (c) the altered cytokine profile in HIV disease, (d) HIV-induced macrophage nitric oxide (NO) production, (e) the increased CD8/CD4 T-cell ratio in HIV infection and (f) the increased colonization of skin by Staphylococcus aureus. However, the observations that (a) LCs cells play an important role in the pathogenesis of psoriasis and a variety of topical and systemic psoriasis treatments cause a reversible decrease in LC function, (b) psoriasis may improve in end-stage HIV infection, (c) overproduction of some TH2 cytokines and underproduction of IL-2 in HIV infection, and (d) the presence of NO favors a TH2 response over a TH1 response make the first four explanations difficult to interpret. Since psoriasis is exacerbated in HIV infection possibly due to the increased staphylococcal colonization, and psoriatic keratinocytes could aggravate HIV infection through production of TNF-alpha, it could be reasoned that in HIV-positive psoriatics a strong vicious cycle is present between the degree of immune deficiency and the staphylococcal colonization, explaining the poor prognosis of both AIDS and psoriasis in these patients. With reference to the studies which indicate significant involvement of substance P (SP) in the pathogenesis of psoriasis and on the other hand increased release of this agent by HIV-infected immune cells it is proposed that SP plays an important role in creating the paradox. Since in HIV-positive psoriatics the source of SP is largely immune cells not neurons, capsaicin, which exerts its action selectively on a subpopulation of neurons, could not be of significant therapeutic value. As SP significantly enhances HIV-1 replication in latently infected immune cells, psoriatic lesions, being heavily infiltrated with immune cells and having high concentrations of SP, could serve as high HIV-replication foci, with the resultant rapid progression of the infection towards AIDS. Additionally, given that lipopolysaccharide is supposed to exacerbate psoriasis, increase of gram negative infections or cutaneous colonization with these organisms in AIDS may partly explain the paradox. Understanding the HIV-induced immunodysregulation that is associated with psoriasis in some HIV-seropositive patients may assist in the delineation of the immunopathogenesis of the disease in HIV-seronegative psoriatics. PMID- 15115315 TI - The beneficial and detrimental effects of linoleic acid on autoimmune disorders. AB - Type 1, or cellular, immune response is characterized by overproduction of IL-1, IL-2, IFN-gamma and TNF-alpha and is the underlying immune mechanism of some autoimmune disorders such as psoriasis, alopecia areata, rheumatoid arthritis, Crohn's disease, multiple sclerosis, insulin-dependent diabetes mellitus and experimental autoimmune uveitis. Type 2 immune response is seen in allergic and antibody-mediated autoimmune diseases and is characterized by IL-4, IL-6 and IL 10 overproduction. Linoleic acid is a precursor of prostaglandin E2 (PGE2) and its intake results in tissue production of PGE2, especially in the absence of other polyunsaturated fatty acids (PUFAS) which inhibit this conversion. PGE2 decreases the production of IL-1, IL-2, IFN-gamma and TNF-alpha and proliferation of TH1 cells and increases the production of IL-4, leading to suppression of the type 1 immune response. Taken together, linoleic acid, the major PUFA of maize oil, could have therapeutic efficacy against cellular autoimmune disorders. On the other hand, excessive intake of linoleic acid may aggravate type 2 autoimmune disorders. PMID- 15115316 TI - The MCP-1 promoter -2518 polymorphism in Behcet's disease: correlation between allele types, MCP-1 production and clinical symptoms among Korean patients. AB - OBJECTIVE: To compare the G vs. A variation at the MCP-1 promoter -2518 position among normal Koreans and Behcet patients, and to investigate possible association of this polymorphism with disease pathogenesis. METHODS: The allele type of -2518 polymorphism was determined by restriction fragment length polymorphism of Pvu II. The level of MCP-1 in serum and culture supernatent was measured by sandwich ELISA. RESULTS: The average serum level of MCP-1 in Behcet patients was 2.37 times higher than in normal controls. The serum MCP-1 concentration was higher in G-allele carriers than in AA homozygotes, and symptoms accompanying graver cases of Behcet's disease, such as gastrointestinal inflammation and uveitis, were more frequent in patients with the G-allele. However, the frequency of G-allele in patient group was not higher than that in healthy Koreans, probably due to the dominance of G-allele in general Korean population. When stimulated in vitro with IL-1beta and LPS, the mononuclear cells from patients carrying the G-allele showed a steeper increase in MCP-1 production than the boost observed in AA homozygotes. CONCLUSION: Although the allele frequency of MCP-1 promoter -2518 polymorphism is not likely to be the reason for the elevated serum MCP-1 level in Korean patients with Behcet's disease, it is possible that proinflammatory factors induced in patients' serum cause stronger activation of MCP-1 expression from the G-type promoter, as well as increased incidence of uveitis and gastric ulcer, among carriers of the G-allele. PMID- 15115317 TI - Increased glutamate release into the intercellular space of the nucleus accumbens (N. accumbens) during substitution of food reinforcement with aversive or neutral stimuli. AB - Experiments on Sprague-Dawley rats using vital intracerebral dialysis and HPLC with electrochemical detection demonstrated that presentation of the animals with inedible imitation food or food containing a bitter flavor instead of the expected food reinforcement led to rapid increases in glutamate levels in the intercellular space of the nucleus accumbens. There was no change in the glutamate level during presentation of the same stimuli to rats not expecting to receive food reinforcement. These data suggest that non-correspondence between the expected and actual values of reinforcement may be an important factor determining the extracellular glutamate level in the nucleus accumbens during food-related behavior. PMID- 15115318 TI - Mathematical simulation of the induction of long-term depression in cerebellar Purkinje cells. AB - The question of the mechanisms underlying the induction of associative and homosynaptic long-term depression in cerebellar Purkinje cells is addressed. Mathematical simulation was used to investigate the possibility that long-term depression, which is associated with a decrease in the efficiency of AMPA receptors, could be induced both by phosphorylation and dephosphorylation of these receptors. PMID- 15115319 TI - The effects of activation of kainate receptors on tonic and phasic gabaergic inhibition in interneurons in field CA1 of guinea pig hippocampus slices. AB - Kainate receptor agonists are powerful convulsants and excitotoxins. Until recently, there have been several contradictory views as to the roles of these receptors in the CNS. We report here experiments showing that application of kainate led to concentration-dependent increases in evoked GABAergic inhibitory postsynaptic currents (phasic currents) in interneurons in field CA1 of guinea pig hippocampus slices. This evidently occurred as a result of a decrease in the action potential generation threshold in inhibitory axons and an increase in the number of endings responding at a given stimulus strength. Increases in phasic inhibitory postsynaptic currents were accompanied by increases in the tonic GABAergic current (the constant component of GABAergic conduction). Increases in the tonic current occurred because of increases in the discharge frequency of interneurons, leading to action-potential-dependent GABA release and, as a result, increases in the extracellular concentration of endogenous agonist. The high level of extracellular GABA after addition of kainate led to desensitization of synaptic GABAergic receptors, while the tonic conductivity led to shunting of synaptic currents. Thus, while 1 microM kainate increased inhibitory postsynaptic currents, this was preceded by a transient depression. The different dynamics of the effects of kainate on phasic and tonic inhibitory GABAergic currents in hippocampal interneurons and the decrease in inhibition of glutamatergic pyramidal cells which may result from these changes may explain the epileptogenic properties of kainate. PMID- 15115320 TI - Immunization of rats with conjugates of dopamine and serotonin with bovine serum albumin prevents the development of experimental MPTP-induced depressive syndrome (electrophysiological parameters). AB - Electrophysiological experiments on Wistar rats demonstrated that prior immunization of animals with conjugates of dopamine and serotonin with bovine serum albumin, as well as with bovine serum albumin alone, played a partial protective role in relation to the subsequent development in these animals of experimental MPTP-induced depressive syndrome: immunized animals showed no signs of the depressive state such as decreases in the latency of onset of REM sleep and the development of epileptiform activity in the caudate-putamen complex, though the increase in the proportion of REM sleep in the overall structure of sleep persisted. Changes in the spectral characteristics of brain electrical activity and sleep structure during the development of experimental MPTP-induced syndrome in animals immunized with conjugates of dopamine and serotonin with bovine serum albumin and with bovine serum albumin alone were antigen-specific and reflected functional shifts in the activity of those neurotransmitter systems targeted by immunization, as well as others sensitive to changes in the body's immunological status. PMID- 15115321 TI - Changes in evoked potentials during the action of sound signals with different localizing characteristics. AB - This report presents results of studies of the phenomenon of mismatch negativity (MMN) during exposure to four blocks of sound stimuli each containing identical standards creating an immobile sound image located along the midline of the head and one of a set of deviants, creating a sound image located either by the left ear or moving from the midline of the head towards the left ear or in the opposite direction. All deviants induced mismatch negativity; the minimal amplitude and longest latent period were seen in the mismatch negativity produced by the deviant modeling movement of the sound image from the midline of the head to the left ear. The question of the appearance of mismatch negativity as a criterion for the accurate discrimination of signals with different localizing characteristics is discussed. PMID- 15115322 TI - Resolving ability and image discretization in the visual system. AB - Psychophysiological studies were performed to measure the spatial threshold for resolution of two "points" and the thresholds for discriminating their orientations depending on the distance between the two points. Data were compared with the scattering of the "point" by the eye's optics, the packing density of cones in the fovea, and the characteristics of the receptive fields of ganglion cells in the foveal area of the retina and neurons in the corresponding projection zones of the primary visual cortex. The effective zone was shown to have to contain a scattering function for several receptors, as this allowed preliminary blurring of the image by the eye's optics to decrease the subsequent (at the level of receptors) discretization noise created by a matrix of receptors. The concordance of these parameters supports the optical operation of the spatial elements of the neural network determining the resolving ability of the visual system at different levels of visual information processing. It is suggested that the special geometry of the receptive fields of neurons in the striate cortex, which are concordant with the statistics of natural scenes, results in a further increase in the signal:noise ratio. PMID- 15115323 TI - The selectivity of neurons in the auditory zone of the mouse midbrain to the direction of movement of a spectral notch in wide-band noise. AB - A series of noise signals was synthesized with spectral notches whose central frequencies moved regularly along the frequency range (from low frequencies to high and from high frequencies to low), imitating movement of the sound source in the vertical plane. The spike responses of neurons in the inferior colliculus of the mouse (Mus musculus) to noise signals changed as the spectral notch moved relative to the excitatory and inhibitory areas of the receptive fields of the neurons and depended on the notch width. Disinhibition reactions in the inhibitory zones were more marked when a frequency notch in the inhibitory zone was followed by a frequency notch in the excitatory part of the response. It is suggested that the selectivity of neurons to the direction of movement of the spectral notch in a noise signal is based on the interaction of the excitatory and inhibitory inputs. The overall set of neuron responses can provide information on the movement of the sound source in acoustic space. PMID- 15115324 TI - Operant behavior in conditions of activation and blockade of neostriatal muscarinic receptors. AB - Chronic experiments were performed on four dogs using a model of an operant defensive reflex associated with maintaining a flexion posture to study the effects of bilateral intraneostriatal microinjection of the non-selective muscarinic receptor agonist carbachol, the selective D2 dopamine receptor blocker raclopride, and the selective M1 muscarinic receptor blocker pirenzipine on the performance of the operant defensive reflex and differentiation of signals. The results show that microinjection of carbachol induced increases in the tonic component and inhibition of the phasic component of the reflex, an ordering rearrangement of the posture, and increases in the amplitudes of its components. Raclopride microinjection gave similar but less marked results. The greatest effects with both substances were seen using differential stimuli. There were sharp increases in the process of differentiation of sound signals. Pirenzipine microinjections gave the opposite result. These data are assessed on the basis of concepts of the existence of two efferent outputs from the neostriatum with opposite effects on their targets and the roles of muscarinic and dopamine receptors in triggering and blocking these effects. PMID- 15115326 TI - The effects of the duration of adaptation to laboratory conditions on the formation of a passive avoidance reflex in rats. AB - The level of adaptation of rats to their new living conditions was studied during formation of a passive avoidance habit using a single combination. A short acclimation period (3 days) had positive influences on the ability of rats to retain a memory trace. There was a negative correlation between step-through latency and measures of anxiety behavior in the elevated cross maze. A change in the adaptation period to nine days decreased the state of anxiety and the level of performance of the conditioned response. There were no correlations between these measures. The modulating role of the level of adaptation to living conditions, associated with different levels of anxiety, in a passive one-session avoidance learning model was assessed. PMID- 15115325 TI - Comparison of the anticonvulsive activities of organic mono- and dications with their abilities to inhibit NMDA and AMPA glutamate receptors. AB - The abilities of mono- and dicationic adamantane and phenylcyclohexyl derivatives to (a) block open NMDA and AMPA glutamate receptors in isolated rat brain neurons and (b) prevent convulsions induced in mice by intraventricular NMDA or kainate were studied. Monocations inhibited NMDA receptors in vitro and produced corresponding protection against NMDA-induced convulsions in vivo, but lacked the ability to block AMPA receptors or prevent kainate-induced convulsions. Dications (IEM-1754 and IEM-1925), which inhibited both NMDA and AMPA receptors, were highly effective at protecting against kainate convulsions and were more effective than the corresponding monocations in preventing NMDA convulsions. The origin of convulsions induced by NMDA appears to be based on a component mediated by activation of AMPA receptors. The anticonvulsive activity of IEM-1754 and IEM 1925 were comparable with those of the known NMDA receptor blockers memantine and MK-801. This was combined with an almost complete absence of the side effects characteristic of memantine and MK-801. The complete correspondence between the in vitro data and in vivo results seen with some of the study compounds is evidently associated with their pharmacokinetic properties. PMID- 15115327 TI - Adaptive behavior in active and passive rats after intranasal administration of corticotrophin-releasing hormone. AB - Rats with high (KHA) and low (KLA) rates of acquiring active avoidance reflexes were used to study the effects of intranasal administration of corticotrophin releasing hormone on orientational-investigative behavior in an open field and anxiety in an elevated cross maze. Administration of the neurohormone induced opposite changes in the behavior of the rats of these lines in the two tests. In KLA rats, movement and investigative activity increased, while in KHA rats these behaviors decreased. In the elevated maze, KLA rats, unlike KHA rats, showed increases in the time spent in the open arms, which was evidence for a decrease in anxiety in these animals. Thus, intranasal hormone administration completely reproduced the effects seen after administration into the striatum. It is suggested that corticotrophin-releasing hormone is an endogenous factor for the detailed and appropriate correction of adaptive behavior. PMID- 15115328 TI - Characteristics of the structure of the speech motor cortex of the frontal area of the brain of a deaf mute child. PMID- 15115329 TI - Expression of the bcl-2 protein in the developing human brain. PMID- 15115330 TI - Formation of the neocortex in rats after prenatal hypoxia. PMID- 15115331 TI - Issues in the diagnosis and treatment of invasive aspergillosis. Introduction. PMID- 15115332 TI - Management of invasive aspergillosis in high-risk patients. AB - Invasive aspergillosis can be difficult to diagnose and control, and conventional drug treatment is often highly toxic, producing medical complications that further compromise patients' health status and escalate health care costs. This article describes the clinical manifestations of Aspergillus infection and discusses approaches to its therapy, including newer pharmaceutical agents with fewer adverse effects, which offer the potential to improve outcomes and substantially lower the cost of treating aspergillosis. PMID- 15115333 TI - Liver fibrosis: from the bench to clinical targets. AB - Progressive liver fibrosis is the main cause of organ failure in chronic liver diseases of any aetiology. Fibrosis develops with different spatial patterns and is a consequence of different prevalent mechanisms according to the diverse causes of parenchymal damage. Indeed, fibrosis, observed as a consequence of chronic viral infection is initially concentrated within and around the portal tract, while fibrosis secondary to toxic/metabolic damage is located mainly in the centrolobular areas. In addition, it is increasingly evident that different cell types are involved in the deposition of fibrillar extracellular matrix during active hepatic fibrogenesis: hepatic stellate cells are mainly involved when hepatocellular damage is limited or concentrated within the liver lobule, whereas portal myofibroblasts and fibroblasts provide a predominant contribution when the damage is located in the proximity of the portal tracts. In the later stages of evolution (septal fibrosis) it is likely that all extracellular matrix producing cells contribute to fibrogenesis. Recruitment and activation of extracellular matrix-producing cells to the site of tissue damage can be due to different major mechanisms: (1) Chronic activation of the tissue repair process. In this case, as a consequence of the reiterated damage, accumulation of fibrillar extracellular matrix reflects the impossibility of an effective remodelling and regeneration. (2) Effect of oxidative stress products, including reactive oxygen intermediates and reactive aldehydes. These products, whose concentration become critical in toxic/metabolic liver injury, are able to induce the synthesis of fibrillar extracellular matrix even in the absence of significant hepatocyte damage and inflammation. (3) Derangement of normal the epithelial/mesenchymal interaction. This typically occurs in all conditions characterised by cholangiocyte damage/proliferation, where a consensual proliferation of extracellular matrix-producing cells and progressive fibrogenesis is commonly observed. A major advancement towards the understanding of the molecular mechanisms of fibrogenesis is derived from a consistent number of in vitro studies investigating the biological role of growth factors/cytokines and other soluble factors and their intracellular signalling pathways. The relevance of these factors has been confirmed by studies performed on animal models and by studies performed on pathological human liver. Along these lines, the elucidation of a consistent number of cellular and molecular mechanisms responsible for the progression of liver fibrosis has provided sound basis for the development of pharmacological strategies able to modulate this important pathophysiological process. Finally, there are several clinically relevant issues that need re-evaluation and/or further investigation, and in particular: (1) the need of an accurate and effective monitoring of the fibrotic progression of chronic liver diseases and of the effectiveness of the currently proposed treatments; (2) the identification of general or individual factors potentially relevant for a faster progression of the disease. PMID- 15115334 TI - Towards the ideal regimen for Helicobacter pylori eradication: the search continues. PMID- 15115335 TI - Predicting the outcome of nonvariceal upper GI bleeding: can we guess right? PMID- 15115336 TI - Endoscopic histology: the start of a new era? PMID- 15115337 TI - Esomeprazole-based therapy in Helicobacter pylori eradication: a meta-analysis. AB - AIM: To perform a systematic review on the efficacy of esomeprazole-based therapies in Helicobacter pylori eradication, and to conduct a meta-analysis comparing the efficacy of esomeprazole and other proton pump inhibitors when co prescribed with antibiotics. METHODS: Studies evaluating esomeprazole plus antibiotics were considered. Only randomised trials comparing esomeprazole and other proton pump inhibitors with antibiotics, and differing only in the proton pump inhibitor, were included in the meta-analysis. Electronic and manual bibliographical searches were conducted. The percentage (weighted mean) of eradication success was calculated. Meta-analysis was performed combining the odd ratios of the individual studies. RESULTS: Mean cure rates with dual regimens (esomeprazole plus clarithromycin) were 51 and 54%, respectively, by intention-to treat and by per-protocol. Corresponding figures with triple regimens (esomeprazole plus clarithromycin and either amoxicillin or metronidazole) were 82% (intention-to-treat) and 86% (per-protocol). Four studies were included in the meta-analysis: mean H. pylori eradication rates (intention-to-treat) with esomeprazole plus antibiotics was 85 and 82% when omeprazole was used (odds ratio = 1.19; 95% confidence interval = 0.81-1.74), results being statistically homogeneous. When subanalysis included only high quality studies, the odds ratio for this comparison was closer to one (1.08; 95% confidence interval = 0.4-1.47) and results were more homogeneous. CONCLUSIONS: Esomeprazole-based triple therapy is highly effective for the eradication of H. pylori infection and offers comparable efficacy to omeprazole-based therapy. PMID- 15115338 TI - Measuring gastric emptying of semisolids in children using the 13C-acetate breath test: a validation study. AB - BACKGROUND AND AIM: Radioscintigraphy is the gold standard for evaluation of gastric emptying in children, but requires exposure to ionising radiation. Therefore, the aim of the study was to validate the non-radioactive 13C-acetate breath test in children in comparison to radioscintigraphy as reference method. PATIENTS: Twenty-nine children with dyspeptic or respiratory symptoms were tested for gastric emptying disorders simultaneously performing the 13C-acetate breath test and radioscintigraphy. METHODS: A semisolid oatmeal was doubly labelled with 150 mg 13C-acetate and 50 MBq 99mTechnetium. Breath samples were collected every 5-10 min for 4 h. After mass spectrometrical 13C-analysis, curve fitting of the 13C-cumulative recovery to the modified power exponential function Y = m(1 - e( kt) calculated the half emptying times of the breath test (t 1/2 (breath)). Scintigraphic image acquisition began immediately after the ingestion of the 99mTechnetium-labelled testmeal at a rate of one frame every 60 s for 1 h. RESULTS: Six children showed delayed gastric emptying in scintigraphy (t 1/2(scinti) > 60 min). All these children had prolonged half emptying times t 1/2 (breath) in the 13C-acetate breath test. Using a cut-off t 1/2(breath) > 90 min, the 13C-acetate breath test had a sensitivity of 100% and a specificity of 85%. Scintigraphic and breath test half emptying times were linearly correlated (Y = 0.80x + 47.68, r = 0.76, P < 0.00001). CONCLUSIONS: The 13C-acetate breath test proves to be a reliable, non-radioactive alternative for measuring gastric emptying in children. PMID- 15115339 TI - Gastric effects of the selective cyclooxygenase-2 inhibitor, celecoxib, in the rat. AB - BACKGROUND: Recent studies have revealed that cyclooxygenase-2 is involved in the protection of the damaged gastric mucosa, mediating, in particular, the acceleration of ulcer healing and angiogenesis; therein, it has been suggested that selective cyclooxygenase-2 inhibitors, although safe in healthy stomach, may have deleterious effects on the injured gastric mucosa. Moreover, no information is available about direct effects of these drugs on gastric surface epithelium. AIMS: To investigate the gastric effects of the selective cyclooxygenase-2 inhibitor, celecoxib, in healthy and damaged rat gastric mucosa. METHODS: Gastric toxicity was studied in the rat by measuring gastric potential difference and mucosal lesions. Celecoxib was administered intragastrically, either in basal conditions or in combination with damaging (acetylsalicylic acid and ethanol) or protective (sodium nitroprusside and lipopolysaccharides from Escherichia coli) agents. The anti-inflammatory activity was evaluated in the carrageenan-induced paw oedema assay. The non-selective inhibitors indomethacin and acetylsalicylic acid were used for comparison. RESULTS: In conscious rats celecoxib, indomethacin and acetylsalicylic acid significantly reduced the paw oedema induced by carrageenan. While acetylsalicylic acid and indomethacin significantly reduced basal gastric potential difference and caused gastric mucosal lesions, celecoxib was ineffective; moreover, it did not aggravate the direct damaging effect of intragastric ethanol or aspirin. Pretreatment with the non-selective nitric oxide synthase inhibitor N-nitro-L-argynine methyl ester did not significantly change the gastric effects of celecoxib. Both celecoxib and indomethacin prevented the gastroprotective effects induced by sodium nitroprusside (nitric oxide donor) or by bacterial lipopolysaccharides (inducer of nitric oxide synthesis). CONCLUSIONS. These data indicate that the selective cyclooxygenase-2 inhibitor celecoxib did not alter gastric mucosal barrier nor induced mucosal lesions in the healthy or nitric oxide-deficient rat gastric mucosa. However, cyclooxygenase 2 inhibition impaired nitric oxide-dependent gastroprotection, indicating that cyclooxygenase-2 derived prostaglandins may be involved in the gastric mucosal defence. PMID- 15115340 TI - Comparison of three different risk scoring systems in non-variceal upper gastrointestinal bleeding. AB - BACKGROUND AND AIMS: To prospectively validate in patients with non-variceal upper gastrointestinal bleeding three risk scoring systems (the Baylor College scoring system, the Rockall's risk scoring system and the Cedars-Sinai Medical Centre predictive index) previously proposed to be predictive of rebleeding/death after upper gastrointestinal bleeding. PATIENTS AND METHODS: We calculated values of the scores for 343 patients, who underwent endoscopy after non-variceal upper gastrointestinal haemorrhage during the years 1997-1999. We compared the observed outcomes with the ones expected upon the original series contributed by the authors. Discriminative ability was evaluated by calculating the area under the receiver operating characteristic curve. RESULTS AND CONCLUSIONS: Rockall's score accurately predicted rebleeding in low- and intermediate-risk categories (< 6), but not in high-risk patients. The rates of rebleeding were significantly higher than the ones predicted by the low-risk categories of either Cedars-Sinai index (< or = 2) or Baylor score (< or = 6). The predicted and the observed mortality was not significantly different throughout all the categories of Rockall's score, except for patients with a score of 4. All the scores had better discriminative ability for mortality than for rebleeding. The Rockall's score identifies a low risk group of patients (Rockall's score < or = 2) for rebleeding and mortality. PMID- 15115341 TI - Treatment of locally advanced hepatocellular carcinoma by hepatic intra-artery chemotherapy: a pilot study. AB - BACKGROUND: Hepatocarcinoma is one of the most common malignant tumours world wide with poor prognosis. Treatment of locally advanced hepatocarcinoma is still controversial. Transcatheter arterial (chemo-)embolisation of hepatocarcinoma are widely used methods but some aspects regarding their use and usefulness have not yet been clarified. Systemic remedies have not yet been proven to affect patient survival. AIMS: To determine if intra-arterial chemotherapy with 5-flurouracil and folinic acid in locally advanced hepatocarcinoma is a viable alternative to existing therapies. PATIENTS: Twenty-four inoperable consecutive patients with locally advanced hepatocarcinoma were enrolled. They all underwent intra-arterial chemotherapy via a surgically implanted port-a-cath, and folinic acid (100 mg/m2) and 5-flurouracil (up to 550 mg/m2) were administered with a 1-week or a 2-week schedule. RESULTS: Nineteen patients completed the study: 2 showed a complete positive response, 11 a partial response, 6 stable disease, while 4 showed a disease progression. Median survival time was 19 (range 4-85) months. Child A patients showed a significant longer survival. CONCLUSIONS: Intra-arterial chemotherapy using folinic acid and 5-flurouracil may be useful in the treatment of locally advanced hepatocarcinoma in cirrhotic patients even in the presence of thrombosis. This treatment could be also useful in comparing transarterial chemoembolisation to a curative treatment. PMID- 15115342 TI - Magnifying gastroendoscopy for diagnosis of histologic gastritis in the gastric antrum. AB - BACKGROUND: We investigated the potential of magnifying endoscopy for diagnosis of histologic gastritis in the gastric antrum. In addition, we investigated whether magnifying endoscopy can be applied for evaluation of Helicobacter pylori eradication therapy. METHODS: We examined 176 Japanese patients including 53 with H. pylori eradication. We evaluated the antrum by magnifying observation and ordinary endoscopic findings, and compared these results. Biopsy specimens were taken from the sites observed. RESULTS: The magnified views were classified into four types. Histology of the biopsy specimens allowed us to match the four magnified views with normal mucosa with fundic glands, normal mucosa with pyloric glands, mucosa with gastritis and intestinal metaplasia/epithelial hyperplasia. The types of magnifying appearances were specific enough for the diagnosis of histologic gastritis (148 out of the 176 (82.4%) cases; sensitivity, 96.3%; specificity, 73.7%). We could accurately diagnose the histologic gastritis by magnifying endoscopy in 49 out of the 53 (92%) cases with H. pylori eradication, while only in 38% by ordinary endoscopy. The accuracy of diagnosis was statistically higher with the use of magnifying endoscopy than with ordinary endoscope (P < 0.001). CONCLUSION: Magnifying gastroendoscopy is useful to judge the histologic gastritis, especially, in cases with H. pylori eradication. PMID- 15115343 TI - Endoscopic finding of granular cell tumour associated with leiomyomas in the oesophagus. AB - Following a single report in the literature of granular cell tumour associated with diffuse leiomyomatosis in the oesophagus, we describe the case of a 39-year old man in whom a granular cell tumour and two leiomyomas were endoscopically removed from this site. This previously unreported association of granular cell tumour with isolated leiomyomas suggests the need to bear in mind the possibility of other mesenchymal lesions, including leiomyomas or leiomyomatosis, when a granular cell tumour is found in the oesophagus. PMID- 15115344 TI - Sepsis and elevated liver enzymes in a patient with inflammatory bowel disease: think of portal vein thrombosis. AB - A 42-year old man, 1 year previously diagnosed with ulcerative colitis after an emergency subtotal colectomy with formation of an ileostomy because of severe colitis with perforation, was admitted with sepsis and jaundice. The liver enzymes were elevated and blood cultures were positive for Streptococcus milleri. Magnetic resonance imaging showed a complete thrombosis of the main stem of the portal vein with occlusion of the left branch. Intravenous antibiotic therapy combined with heparinisation led to complete recanalisation of the thrombus. Portal vein thrombosis is a rare complication of inflammatory bowel disease and has been described in only 10 patients thus far. Multiple aetiologic factors may be responsible in relation to inflammatory bowel disease, such as hypercoagulability, thrombocytosis and abdominal sepsis. In patients with inflammatory bowel disease, unexplained sepsis and abnormal liver function tests, the possibility of an acute portal vein thrombosis should be considered and investigated, because unrecognised it may have serious long-term complications. PMID- 15115345 TI - Celebrating our value. PMID- 15115346 TI - News flash nursing shortage is over! PMID- 15115348 TI - Home study program. Can we build a safer OR? AB - PEOPLE WHO WORK in health care are among the brightest and most dedicated workers in the United States, but they are human, and humans make mistakes. HEALTH CARE FACILITIES are moving away from a culture of perfection and exploring how human factors predispose people to make certain types of errors. THIS ARTICLE discusses the types of errors being made and the organizations that are working to redesign the health care system to make it easier to do the job more safely and more difficult to make a mistake. PMID- 15115349 TI - Keeping patients safe--procedure and site verification and preprocedure pause. AB - PATIENT SAFETY GOALS are part of new policies issued by the Joint Commission on Accreditation of Healthcare Organizations. THESE NEW POLICIES led Inova Health System, Falls Church, Va, to develop a standardized, system-wide policy for procedure verification and a preprocedure pause. THE MULTIFACILITY COMPANY overcame a variety of obstacles to implement a system that could be used across all departments in five hospitals. PMID- 15115350 TI - Code red in the OR----implementing an OR fire drill. AB - KNOWLEDGE OF FIRE PREVENTION and fire safety is essential for every OR staff member. In the event of a surgical fire, the quick and knowledgeable response of surgical team members will affect their own safety and that of their patients. A NEW FIRE SAFETY PLAN was developed at Peninsula Regional Medical Center, Salisbury Md. To update staff members' knowledge of the facility's fire plan and special instructions for the OR, a comprehensive educational program on fire safety was presented, and an OR fire drill was conducted. THIS ARTICLE describes the facility's quest to improve knowledge of fire safety by providing multiple educational opportunities. PMID- 15115351 TI - Case outsourcing medical device reprocessing. AB - IN THE INTEREST OF SAVING MONEY, many hospitals are considering extending the life of some single-use medical devices by using medical device reprocessing programs. FACILITIES OFTEN LACK the resources required to meet the US Food and Drug Administration's tough quality assurance standards. BY OUTSOURCING, hospitals can reap the benefits of medical device reprocessing without assuming additional staffing and compliance burdens. OUTSOURCING enables hospitals to implement a medical device reprocessing program quickly, with no capital investment and minimal effort. PMID- 15115352 TI - The invisible nurse--behind the scenes in an Australian OR. AB - THE DWINDLING NUMBER of RNs choosing to work in the OR has been caused by many factors, such as restricted undergraduate exposure to the perioperative experience and fewer postgraduate opportunities. PRESSURE TO IDENTIFY alternative health care workers to replace RNs in the OR is escalating to a point where the role of the perioperative RN, as currently understood, is threatened. FINDINGS from an Australian ethnographic study suggest that both patients and non-nursing colleagues have limited appreciation of the contribution that perioperative nurses make. THE NEED FOR NURSES to take a proactive stance on this subject is discussed within the context of professional and structural change. PMID- 15115353 TI - The value of certification--a research journey. AB - ONE MEASURE OF NURSING EXCELLENCE is achieving specialty certification, which denotes a more advanced level of knowledge and practice. THE CERTIFICATION BOARD Perioperative Nursing (CBPN) provides specialty certification for perioperative nurses. The organization's mission is to be the leader in competency credentialing and education that promotes safe, quality patient care in the perioperative arena. THIS ARTICLE PRESENTS study findings regarding the perceived value of a barriers to obtaining certification. PMID- 15115354 TI - Recommended practices for laser safety in practice settings. PMID- 15115355 TI - Stress management through the viewfinder. PMID- 15115356 TI - A strategic view of AORN's federal legislative issues. PMID- 15115357 TI - Safety resources for perioperative clinicians. PMID- 15115358 TI - Coming out on top. PMID- 15115359 TI - An evaluation of neuroscience nursing research published during the Decade of the Brain. AB - The purpose of this study was to extend a previous evaluation of neuroscience nursing research to an assessment of the research published during the 1990s--the Decade of the Brain. The evaluation was conducted to assess the focus of neuroscience nursing research and to identify research strategies and scientific methods. The Journal of Neuroscience Nursing, Nursing Research, Research in Nursing & Health, and Western Journal of Nursing Research were canvassed. An assessment form developed by the authors was used to evaluate selected articles published between January 1989 and December 2000. The number of neuroscience research studies published in nursing journals increased substantially during the 1990s. The studies focused more on individuals with neurological disorders and less on neurological trauma. The findings demonstrated major changes in subspecialty area, care orientation, and nature of study variables from that observed in the previous evaluation in which acute care and physiological variables were most representative of the pre-1989 studies. As noted in the previous evaluation, the trend toward theory-then-research studies continued and the scientific methods grew more complex. PMID- 15115360 TI - Withholding aggressive seizure treatment from a child with severe intractable epilepsy: a case study. AB - A child with severe intractable epilepsy, profound neurological impairment, and frequent and lengthy admissions to the hospital for repeated and prolonged status epilepticus has an uncertain quality of life. The benefit and ethics of repetitive, aggressive seizure treatment are in question. This case study describes a healthcare team's experience when it was decided to withhold aggressive seizure treatment from a child with severe intractable epilepsy and transition to a palliative care approach. The situation leading up to this decision and the ethical dilemmas, nursing issues and interventions, and lessons learned are reviewed. PMID- 15115361 TI - Love and load--the lived experience of the mother-child relationship among young adult traumatic brain-injured survivors. AB - This study aims to describe the meaning of the experience of the relationship between young adult traumatic brain injury (TBI) survivors and their mothers using a phenomenological approach. Informants included 9 males and 3 females who were at least 2 years post-TBI, and their mothers, who were their primary caregivers after the injury. TBI informants were 18 to 25 years of age, had motor vehicle accident-induced injury, experienced post-traumatic amnesia longer than 24 hours, and were able to participate in a verbal interview. In addition, all informants currently were living with their mothers, who also participated in this study. Survivors acquired the sense of being abnormal from various sources, including social pressures, dynamics within the family, and intrapersonal changes. Mothers adopted both positive and negative actions during the period of uncertainty and often struggled to balance protecting their children and letting them become independent. They also struggled to maintain harmonious relationships with people both inside and outside of the family. Sometimes, survivors' parents marital relationships were at risk. Health professionals should design more appropriate long-term community interventions to help TBI survivors and their families decrease the burden of injury and the resulting stress, increase survivors' self-esteem, and improve quality of life of both survivors and their families, serving as a foundation for further TBI care. PMID- 15115362 TI - Chronic sorrow and depression in parents of children with neural tube defects. AB - Chronic sorrow has been described in the caregivers of individuals with myelomeningocele, multiple sclerosis, Parkinson's disease, cancer, and premature infants. Most studies have reported small numbers of fathers for comparison with mothers. One unpublished study compared chronic sorrow and depression. The purpose of the current study was to describe parental chronic sorrow following the birth of a child with neural tube defect and to explore the relationship between chronic sorrow and depression. One hundred and thirty-two parents (63 mother/father pairs and 6 single parents) responded to 3 measures of chronic sorrow and 1 measure of depression. Multivariate analysis of variance demonstrated significant differences between mothers and fathers in chronic sorrow. In addition, a relationship between chronic sorrow and depression was demonstrated. Chronic sorrow is a potential barrier to parental understanding of their child's care and diagnosis. Based on these findings, separate assessments of each parent and timely interventions are warranted. Further research should include more than 1 measure of chronic sorrow and delineate the dimensions being measured. PMID- 15115363 TI - 2-year-old with tuberculous meningitis: a case study. AB - Tuberculous meningitis (TBM) may occur with tuberculosis infection, and young children are more prone to this disease. The clinical manifestations, time course, and treatment of TBM are unlike those of other types of meningitis, and the disease presents unique challenges for nurses caring for these patients. This case study highlights the typical presentation, course, and management of TBM in a pediatric patient and provides an overview of this devastating disease. Specific nursing issues related to the care of these children are outlined. PMID- 15115364 TI - Time and difficulty of tasks provided by family caregivers of stroke survivors. AB - Family caregivers of stroke survivors are at risk for negative health outcomes such as depression, psychosocial impairments, and even mortality as a result of providing care. Shortened hospital stays have contributed to the urgent need for caregivers to manage difficult and time-consuming tasks required for the care of stroke survivors in the home setting. The purposes of this study were to (a) identify which tasks were perceived as most time-consuming and difficult, (b) determine which of these tasks were most predictive of mood and other negative caregiver outcomes, and (c) evaluate the psychometric properties of the Oberst Caregiving Burden Scale (OCBS) as a measure of tasks in stroke caregivers. A cross-sectional design was employed using mailed questionnaires from 116 family caregivers of stroke survivors featuring the OCBS, the Profile of Mood States Short Form, and the Bakas Caregiving Outcomes Scale. The tasks perceived as most time-consuming and difficult, which also were predictive of mood and other negative caregiver outcomes, were managing finances, managing behaviors, and providing emotional support. Strong psychometric properties of the OCBS were found at both item and subscale levels. Developing an individual profile using the OCBS items may help to target individual support interventions for caregivers. Suggested interventions include referring caregivers to resources for tasks that fall outside the scope of nursing practice, supporting caregivers with tasks such as providing emotional support or managing behavioral problems, and encouraging caregivers to seek care for their own physical, emotional, or social needs. PMID- 15115365 TI - A new understanding. PMID- 15115366 TI - Response to erroneous comments made in Sanhueza et al. PMID- 15115367 TI - Recycling in a megacity. AB - In the aftermath of the 9/11 disaster, Mayor Bloomberg of New York City unveiled an aggressive budget plan that included the temporary suspension of glass and plastics recycling. This was considered by many to be anti-environmental, but the results of this study show that for lack of markets, even at zero or negative prices, nearly 90% of the plastic and glass set aside by thoughtful New Yorkers was transported to materials recovery facilities (MRFs) and from there to landfills. Sending bales of plastics to landfills is not limited to New York City. It is an environmental paradox that the United States is digging up new oil fields in pristine areas and, at the same time, continues to convert greenfields to brownfields by burying nearly 20 million tons of plastic fuel annually. The study also determined that at the present rate of source separation, estimated to be less than 30% of the available recyclables in 1999, building large, modern MRFs may increase substantially the rate of New York City recycling and also allow single-stream collection of commingled recyclables, as is done in Phoenix, AZ. Single-stream collection simplifies separation at the source by citizens and increases the amount of collected recyclables. Also, because collection represents a large fraction of the costs of waste management, it may have a significant economic advantage. PMID- 15115368 TI - Evaluation of the Industrial Source Complex Short-Term model: dispersion over terrain. AB - Terrain around an air discharge source can have several influences on diffusion, the pattern of plume dispersion, the wind flow, and the turbulence characteristics. The Industrial Source Complex Short-Term (ISCST) model contains simple algorithms to attempt to account for the effects of terrain. The model has the ability to analyze concentrations in any type of terrain by using the terrain options available for running the model. In this study, the ISCST model was adopted to predict the concentration of sulfur dioxide (SO2) in and around the Mina Al-Fahal refinery in Oman. The central purpose of the study was to examine the performance of the ISCST model in predicting SO2 concentrations under two different scenarios: (1) when flat terrain was assumed; and (2) when the terrain descriptions were addressed. The results of these two scenarios were validated against SO2 monitoring data. The comparison showed that the model underestimated the observed concentrations for the two scenarios. However, the predicted concentrations of SO2 in the absence of the terrain scenario were in better agreement with the observations. Furthermore, the predicted SO2 concentrations were found to be lower than the World Health Organization guideline values, with the maximum concentrations found to occur relatively close to the sources of emission. PMID- 15115369 TI - Effect of substrate Henry's constant on biofilter performance. AB - Butanol, ether, toluene, and hexane, which have Henry's constants ranging from 0.0005 to 53, were used to investigate the effects of substrate solubility or availability on the removal of volatile organic compounds (VOCs) in trickle-bed biofilters. Results from this study suggest that, although removal of a VOC generally increases with a decrease in its Henry's constant, an optimal Henry's constant range for biofiltration may exist. For the treatment of VOCs with high Henry's constant values, such as hexane and toluene, the transfer of VOCs between the vapor and liquid phases or between the vapor phase and the biofilm is a rate determining step. However, oxygen (O2) transfer may become a rate-limiting step in treating VOCs with low Henry's constants, such as butanol, especially at high organic loadings. The results demonstrated that in a gas-phase aerobic biofilter, nitrate can serve both as a growth-controlling nutrient and as an electron acceptor in a biofilm for the respiration of VOCs with low Henry's constants. Microbial communities within the biofilters were examined using denaturing gradient gel electrophoresis to provide a more complete picture of the effect of O2 limitation and denitrification on biofilter performance. PMID- 15115370 TI - Field validation of an active sampling cartridge as a passive sampler for long term carbonyl monitoring. AB - A carbonyl sampler originally designed for the active sampling method (Sep-Pak XPoSure) was used for long-term passive sampling, and its applicability as a passive sampler was examined through field experiments. The uptake rates of passive sampling were determined experimentally from collocated passive and active samplings for various sampling periods. The obtained uptake rates of formaldehyde, acetaldehyde, and acetone were 1.48, 1.23, and 1.08 mL/min, respectively. These uptake rates were consistent for a wide range of the sampling term (12 hr-2 weeks). Uptake rates of each carbonyl were proportional to the diffusion coefficients of each. Therefore, the ratios of diffusion coefficients were used to calculate the uptake rates of carbonyls for which the rates were not determined experimentally. Lower limits of determination were 2.16-17.5 microg/m3 for 2-week sampling. It was confirmed that 2-week monitoring of carbonyl concentrations up to 118-229 microg/m3 was possible. Relative standard deviations of the passive method generated from the repeatability test were 2-12.3% error for five samplings, and the recovery efficiencies were larger than 90%. Thus, the passive sampler was found to be highly suitable for long-term monitoring of carbonyl compounds. PMID- 15115371 TI - Source characterization of atmospheric heavy metals in industrial/residential areas: a case study in Oman. AB - Total suspended particulate matter (TSP) was collected from two districts: the Sohar industrial estate (SIE) and a residential area in the case study area in Oman. The TSP collected from SIE was taken from 19 different industrial activities, whereas those collected from the residential area were taken from 12 houses. The samples were analyzed for nine heavy metals: chromium (Cr), copper (Cu), manganese (Mn), nickel, lead (Pb), zinc (Zn), cadmium, vanadium, and molybdenum. The results were used to assess the source classes responsible for TSP and heavy metal levels in the atmosphere of the two districts. At each district, principal component analysis was applied to the concentrations of TSP and heavy metals to obtain the number of principal components. At SIE, three groups of elements were determined. The first component contained large factor loadings in Cu and Mn (Cu industrial processes and reinforcement steel production). The second presented most of the variance of Cr, Pb, and Zn (black and galvanized iron pipes production, mechanical industries, and vehicle construction). The third component was composed of Zn and Pb that probably reflected contributions from motor-vehicle tire wear or was associated with municipal incineration. On the other hand, results from the village showed that only one factor was able to explain the main part of the data variance, and the industrial site was polluting air quality in the village with Cu, Cr, Pb, nickel, and Mn. PMID- 15115372 TI - Atmospheric concentrations of PM2.5 trace elements in the Seoul urban area of South Korea. AB - Fine particles (PM2.5) were collected during all four seasons, from April 2001 to February 2002, in Seoul, South Korea, using an annular denuder system. Elemental compositions of ambient PM2.5 were analyzed using the proton-induced X-ray emission method. The greatest contributors (> or = 2%) to the PM2.5 mass were sulfur (S), silicon (Si), chlorine (Cl), aluminum (Al), and iron (Fe) in the spring; S in the summer; and S and Cl in the fall. S, Cl, and Si were the major elements in the winter. S was the most abundant species among the elements, ranging from 5.3 to 7.9%, followed by Si and Cl. From analysis of variance, PM2.5 mass, Al, Si, potassium, calcium, and Fe showed significant seasonal differences during the four seasons (p < 0.001). Enrichment factor (EF) analysis was carried out to identify the sources affecting the aerosol in the Seoul area. On the basis of the mean EF values, elemental S, copper, zinc, and lead may be emitted from anthropogenic sources (EF > 50). Elemental Al, Si, titanium, and Fe may be emitted from crustal sources (EF < 3). Additionally, a correlation analysis was carried out for source identification. The results of the correlation analysis were confirmed by the results of the EF analysis. PMID- 15115373 TI - Spatial variability of PM2.5 in urban areas in the United States. AB - Data from the U.S. Environmental Protection Agency's Aerometric Information Retrieval System (now known as the Air Quality System) database for 1999 and 2000 have been used to characterize the spatial variability of concentrations of particulate matter with aerodynamic diameter < or = 2.5 microg (PM2.5) in 27 urban areas across the United States. Different measures were used to quantify the degree of uniformity of PM2.5 concentrations in the urban areas characterized. It was observed that PM2.5 concentrations varied to differing degrees in the urban areas examined. Analyses of several urban areas in the Southeast indicated high correlations between site pairs and spatial uniformity in concentration fields. Considerable spatial variation was found in other regions, especially in the West. Even within urban areas in which all site pairs were highly correlated, a variable degree of heterogeneity in PM2.5 concentrations was found. Thus, even though concentrations at pairs of sites were highly correlated, their concentrations were not necessarily the same. These findings indicate that the potential for exposure misclassification errors in time-series epidemiologic studies exists. PMID- 15115374 TI - Operational characteristics of effective removal of H2S and NH3 waste gases by activated carbon biofilter. AB - Simultaneous removal of hydrogen sulfide (H2S) and ammonia (NH3) gases from gaseous streams was studied in a biofilter packed with granule activated carbon. Extensive studies, including the effects of carbon (C) source on the growth of inoculated microorganisms and gas removal efficiency, product analysis, bioaerosol emission, pressure drop, and cost evaluation, were conducted. The results indicated that molasses was a potential C source for inoculated cell growth that resulted in removal efficiencies of 99.5% for H2S and 99.2% for NH3. Microbial community observation by scanning electron microscopy indicated that granule activated carbon was an excellent support for microorganism attachment for long-term waste gas treatment. No disintegration or breakdown of biofilm was found when the system was operated for 140 days. The low bioaerosol concentration emitted from the biofilter showed that the system effectively avoided the environmental risk of bioaerosol emission. Also, the system is suitable to apply in the field because of its low pressure drop and treatment cost. Because NH3 gas was mainly converted to organic nitrogen, and H2S gas was converted to elemental sulfur, no acidification or alkalinity phenomena were found because of the metabolite products. Thus, the results of this study demonstrate that the biofilter is a feasible bioreactor in the removal of waste gases. PMID- 15115375 TI - Temporal variation in daily concentrations of ozone and acid-related substances at Saturna Island, British Columbia. AB - A multiple linear regression model was used to investigate seasonal and long-term trends in concentrations of ozone (O3) and acid-related substances at the Saturna Island monitoring station in southwestern British Columbia from 1991 to 2000. Statistically significant primary (dominant) cycles with a period of 1 yr were found for O3, sulfur dioxide (SO2), nitric acid (HNO3), and aerosol concentrations of sulfate (SO4(2-)), calcium (Ca2+) and chloride (Cl-). Of these, peak median concentrations occurred during the spring for O3 and Ca2+, during the warmer, drier months (April-September) for SO4(2-) and HNO3, and during the cooler, wetter months (October-March) for SO2 and Cl-. Statistically significant secondary cycles of 6 months duration were seen for concentrations of O3, SO4(2 ), HNO3, Ca2+, and Cl-. Daily maximum O3 concentrations exhibited a statistically significant increase over the period of record of 0.33 +/- 0.26 ppb/yr. Statistically significant declines were found for concentrations of SO2, SO4(2-), HNO3, Ca2+, and potassium, ranging from 20 to 36% from levels at the start of the sampling period. Declines in ambient concentrations of SO2, SO4(2-), and HNO3 reflect local declines in anthropogenic emissions of the primary precursors SO2 and NOx over the past decade. Trends in Ca2+ and potassium ion concentrations are in line with a broader North American declining trend in acid-neutralizing cations. PMID- 15115376 TI - Near-field dispersion modeling for regulatory applications. AB - This paper evaluates the application of dispersion models to estimate near-field pollutant concentrations in two case studies. The Industrial Source Complex Short Term Model (ISCST3) was evaluated with hexavalent chromium measurements collected within 100 m of two facilities in Barrio Logan, San Diego, CA. ISCST3 provided reasonable estimates for higher pollutant concentrations but underestimated lower concentrations. To understand the observed distribution of concentrations in Barrio Logan, a recently conducted tracer experiment was analyzed. The tracer, sulfur hexafluoride, was released at ambient temperature from an urban facility at the University of California at Riverside, and concentrations were measured within 20 m of the source. Modeling results indicated that Industrial Source Complex-Plume Rise Model Enhancement and American Meteorological Society/U.S. Environmental Protection Agency Regulatory Model-Plume Rise Model Enhancement overestimated high concentrations and underestimated low concentrations. A diagnostic study with a simple Gaussian dispersion model that incorporated site specific meteorology was used to evaluate model results. This study found that incorporating lateral meandering for nonbuoyant urban plumes in Gaussian dispersion models could improve concentration estimates even when downwash is not considered. Incorporating a meandering component in ISCST3 resulted in improvements in estimating hexavalent chromium concentrations in Barrio Logan. Credible near-source concentration estimates depend on accurate characterization of emissions, onsite micrometeorology, and a method to account for lateral meandering in the near field. PMID- 15115377 TI - Prediction of the vertical profile of ozone based on ground-level ozone observations and cloud cover. AB - A number of statistical techniques have been used to develop models to predict high-elevation ozone (O3) concentrations for each discrete hour of day as a function of elevation based on ground-level O3 observations. The analyses evaluated the effect of exclusion/inclusion of cloud cover as a variable. It was found that a simple model, using the current maximum ground-level O3 concentration and no effect of cloud cover provided a reasonable prediction of the vertical profile of O3, based on data analyzed from O3 sites located in North Carolina and Tennessee. The simple model provided an approach that estimates the concentration of O3 as a function of elevation (up to 1800 m) based on the statistical results with a +/- 13.5 ppb prediction error, an R2 of 0.56, and an index of agreement, d1, of 0.66. The inclusion of cloud cover resulted in a slight improvement in the model over the simple regression model. The developed models, which consist of two matrices of 24 equations (one for each hour of day for clear to partly cloudy conditions and one for cloudy conditions), can be used to estimate the vertical O3 profile based on the inputs of the current day's 1-hr maximum ground-level O3 concentration and the level of cloud cover. PMID- 15115378 TI - A risk-based model to establish threshold planning quantities of hazardous substances. AB - The Threshold Planning Quantity (TPQ) is the minimum quantity of any hazardous substance stored or processed at an industry that can pose a defined level of risk up to a certain distance from the industrial premises should an accidental release occur. These are recommended by the concerned government departments of the respective countries. Often, concerned regulatory authorities focus on simple screening tools to prescribe TPQs of hazardous chemicals rather than on more detailed technical aspects. This is illustrated with the help of TPQs presently administered in India, which are followed based on Control of Industrial Major Accident Hazard regulations of the United Kingdom. In this paper, the risk ranking matrix (RRM) approach is described to evaluate the recommended TPQs and prescribe these for some extremely hazardous chemicals (EHS) commonly used in industries. The RRM presented here is unique in the sense that various acceptable risk criteria are reviewed to appropriately rank the risk potential and provide suitability criteria in terms of individual risk factors and geosocietal risk factors (GSRF). Based on this concept, an attempt has been made to develop a risk based model to determine the TPQs of various EHS (acrolein, ammonia, chlorine, hydrogen fluoride, and phosgene). PMID- 15115379 TI - Lessons learned from marketed and investigational prodrugs. PMID- 15115380 TI - Piperazine-based CCR5 antagonists as HIV-1 inhibitors. IV. Discovery of 1-[(4,6 dimethyl-5-pyrimidinyl)carbonyl]- 4-[4-[2-methoxy-1(R)-4 (trifluoromethyl)phenyl]ethyl-3(S)-methyl-1-piperazinyl]- 4-methylpiperidine (Sch 417690/Sch-D), a potent, highly selective, and orally bioavailable CCR5 antagonist. AB - The nature and the size of the benzylic substituent are shown to be the key to controlling receptor selectivity (CCR5 vs M1, M2) and potency in the title compounds. Optimization of the lead benzylic methyl compound 3 led to the methoxymethyl analogue 30, which had excellent receptor selectivity and oral bioavailability in rats and monkeys. Compound 30 (Sch-417690/Sch-D), a potent inhibitor of HIV-1 entry into target cells, is currently in clinical trials. PMID- 15115381 TI - Synthesis and evaluation of oxazaborolidines for antibacterial activity against Streptococcus mutans. AB - Several representative oxazaborolidines have been synthesized and evaluated against S. mutans for antibacterial activity. This is the first reported antibacterial activity of this class of compounds. The minimal inhibitory concentration values ranged from 0.53 to 6.75 mM. PMID- 15115382 TI - Development of irreversible diphenyl phosphonate inhibitors for urokinase plasminogen activator. AB - In this letter we report the synthesis and biochemical evaluation of selective, irreversible diphenyl phosphonate inhibitors for urokinase plasminogen activator (uPA). A diphenyl phosphonate group was introduced on the substratelike peptide Z d-Ser-Ala-Arg, and modification of the guanidine side chain was investigated. A guanylated benzyl group appeared the most promising side chain modification. A k(app) value in the 10(3) M(-1) s(-1) range for uPA was obtained, together with a selectivity index higher than 240 toward other trypsin-like proteases such as tPA, thrombin, plasmin, and FXa. PMID- 15115383 TI - Identification of 4-(1H-imidazol-4(5)-ylmethyl)pyridine (immethridine) as a novel, potent, and highly selective histamine H(3) receptor agonist. AB - In this study, the piperidine ring of immepip and its analogues was replaced by a rigid heterocyclic pyridine ring. Many compounds in the series exhibit high affinity and agonist activity at the human histamine H(3) receptor. Particularly, the 4-pyridinyl analogue of immepip (1c, immethridine) is identified as a novel potent and highly selective histamine H(3) receptor agonist (pK(i) = 9.07, pEC(50) = 9.74) with a 300-fold selectivity over the closely related H(4) receptor. PMID- 15115384 TI - Discovery of diarylacrylonitriles as a novel series of small molecule sortase A inhibitors. AB - On the basis of a hit from random screening, a novel class of small-molecule sortase A inhibitors was generated. The primary structure-activity relationship and the minimal structural requirements for potency were established through structural modifications and molecular modeling studies. PMID- 15115385 TI - Design and synthesis of alpha-aryloxy-alpha-methylhydrocinnamic acids: a novel class of dual peroxisome proliferator-activated receptor alpha/gamma agonists. AB - The design and synthesis of the dual peroxisome proliferator activated receptor (PPAR) alpha/gamma agonist (S)-2-methyl-3-[4-[2-(5-methyl-2-thiophen-2-yl-oxazol 4-yl)ethoxy]phenyl]-2-phenoxypropionic acid (2) for the treatment of type 2 diabetes and associated dyslipidemia are described. 2 possesses a potent dual hPPAR alpha/gamma agonist profile (IC(50) = 28 and 10 nM; EC(50) = 9 and 4 nM, respectively, for hPPARalpha and hPPARgamma). In preclinical models, 2 substantially improves insulin sensitivity and potently reverses diabetic hyperglycemia while significantly improving overall lipid homeostasis. PMID- 15115386 TI - N-phenylphenylglycines as novel corticotropin releasing factor receptor antagonists. AB - Screening of a computationally designed synthetic library led to the discovery of the N-phenylphenylglycines (NPPGs) as a novel class of human corticotropin releasing factor (h-CRF(1)) antagonists. Several NPPGs with greater potency than the original hit 1 were rapidly identified, and resolution of the racemate demonstrated that only the R-enantiomer displays activity. This structural class represents the first example of a non-peptide CRF(1) antagonist with a stereochemically distinct receptor binding affinity. PMID- 15115387 TI - Discovery of embelin as a cell-permeable, small-molecular weight inhibitor of XIAP through structure-based computational screening of a traditional herbal medicine three-dimensional structure database. AB - The X-linked inhibitor of apoptosis (XIAP) is a promising new molecular target for the design of novel anticancer drugs aiming at overcoming apoptosis resistance of cancer cells to chemotherapeutic agents and radiation therapy. Recent studies demonstrated that the BIR3 domain of XIAP where caspase-9 and Smac proteins bind is an attractive site for designing small-molecule inhibitors of XIAP. Through computational structure-based screening of an in-house traditional herbal medicine three-dimensional structure database of 8221 individual natural products, followed by biochemical testing of selected candidate compounds, we discovered embelin from the Japanese Ardisia herb as a small-molecular weight inhibitor that binds to the XIAP BIR3 domain. We showed that embelin binds to the XIAP BIR3 protein with an affinity similar to that of the natural Smac peptide using a fluorescence polarization-based binding assay. Our NMR analysis further conclusively confirmed that embelin interacts with several crucial residues in the XIAP BIR3 domain with which Smac and caspsase-9 bind. Embelin inhibits cell growth, induces apoptosis, and activates caspase-9 in prostate cancer cells with high levels of XIAP, but has a minimal effect on normal prostate epithelial and fibroblast cells with low levels of XIAP. In stably XIAP-transfected Jurkat cells, embelin effectively overcomes the protective effect of XIAP to apoptosis and enhances the etoposide-induced apoptosis and has a minimal effect in Jurkat cells transfected with vector control. Taken together, our results showed that embelin is a fairly potent, nonpeptidic, cell-permeable, small-molecule inhibitor of XIAP and represents a promising lead compound for designing an entirely new class of anticancer agents that target the BIR3 domain of XIAP. PMID- 15115388 TI - Novel N-arylpyrazolo[3,2-c]-based ligands for the glucocorticoid receptor: receptor binding and in vivo activity. AB - A novel series of selective ligands for the human glucocorticoid receptor (hGR) are described. Preliminary structure-activity relationships were focused on substitution at C-1 and indicated a preference for 3-, 4-, and 5-substituted aromatic and benzylic groups. The resulting analogues, e.g., 18 and 34, exhibited excellent affinity for hGR (IC(50) 1.9 nM and 2.8 nM, respectively) and an interesting partial agonist profile in functional assays of transactivation (tyrosine aminotransferase, TAT, and glutamine synthetase, GS) and transrepression (IL-6). The most potent compounds described in this study were the tertiary alcohol derivatives 21 and 25. These candidates showed highly efficacious IL-6 inhibition versus dexamethasone. The thiophenyl analogue 25 was evaluated in vivo in the mouse LPS challenge model and showed an ED(50) = 4.0 mg/kg, compared to 0.5 mg/kg for prednisolone in the same assay. PMID- 15115389 TI - 5-substituted derivatives of 6-halogeno-3-((2-(S)-azetidinyl)methoxy)pyridine and 6-halogeno-3-((2-(S)-pyrrolidinyl)methoxy)pyridine with low picomolar affinity for alpha4beta2 nicotinic acetylcholine receptor and wide range of lipophilicity: potential probes for imaging with positron emission tomography. AB - Potential positron emission tomography (PET) ligands with low picomolar affinity at the nicotinic acetylcholine receptor (nAChR) and with lipophilicity (log D) ranging from -1.6 to +1.5 have been synthesized. Most members of the series, which are derivatives of 5-substituted-6-halogeno-A-85380, exhibited a higher binding affinity at alpha4beta2-nAChRs than epibatidine. An analysis, by molecular modeling, revealed an important role of the orientation of the additional heterocyclic ring on the binding affinity of the ligands with nAChRs. The existing nicotinic pharmacophore models do not accommodate this finding. Two compounds of the series, 6-[(18)F]fluoro-5-(pyridin-3-yl)-A-85380 ([(18)F]31) and 6-chloro-3-((2-(S)-azetidinyl)methoxy)-5-(2-[(18)F]fluoropyridin-5-yl)pyridine) ([(18)F]35), were radiolabeled with (18)F. Comparison of PET data for [(18)F]31 and 2-[(18)F]FA shows the influence of lipophilicity on the binding potential. Our recent PET studies with [(18)F]35 demonstrated that its binding potential values in Rhesus monkey brain were ca. 2.5 times those of 2-[(18)F]FA. Therefore, [(18)F]35 and several other members of the series, when radiolabeled, will be suitable for quantitative imaging of extrathalamic nAChRs. PMID- 15115390 TI - Reducing the peptidyl features of caspase-3 inhibitors: a structural analysis. AB - Caspases are cysteine proteases that specifically cleave Asp-Xxx bonds. They are key agents in inflammation and apoptosis and are attractive targets for therapy against inflammation, neurodegeneration, ischemia, and cancer. Many caspase structures are known, but most involve either peptide or protein inhibitors, unattractive candidates for drug development. We present seven crystal structures of inhibited caspase-3 that illustrate several approaches to reducing the peptidyl characteristics of the inhibitors while maintaining their potency and selectivity. The inhibitors reduce the peptidyl nature of inhibitors while preserving binding potency by (1). exploiting a hydrophobic binding site C terminal to the cleavage site, (2). replacing the negatively charged aspartyl residue at P4 with neutral groups, and (3). using a peptidomimetic 5,6,7 tricyclic system or a pyrazinone at P2-P3. In addition, we have found that two nicotinic acid aldehydes induce a significant conformational change in the S2 and S3 subsites of caspase-3, revealing an unexpected binding mode. These results advance the search for caspase-directed drugs by revealing how unacceptable molecular features can be removed without loss of potency. PMID- 15115391 TI - Synthesis of 2,4-diamino-6-[2'-O-(omega-carboxyalkyl)oxydibenz[b,f]azepin-5 yl]methylpteridines as potent and selective inhibitors of Pneumocystis carinii, Toxoplasma gondii, and Mycobacterium avium dihydrofolate reductase. AB - Six previously undescribed N-(2,4-diaminopteridin-6-yl)methyldibenz[b,f]azepines with water-solubilizing O-carboxyalkyloxy or O-carboxybenzyloxy side chains at the 2'-position were synthesized and compared with trimethoprim (TMP) and piritrexim (PTX) as inhibitors of dihydrofolate reductase (DHFR) from Pneumocystis carinii (Pc), Toxoplasma gondii (Tg), and Mycobacterium avium (Ma), three of the opportunistic organisms known to cause significant morbidity and mortality in patients with AIDS and other disorders of the immune system. The ability of the new analogues to inhibit reduction of dihydrofolate to tetrahydrofolate by Pc, Tg, Ma, and rat DHFR was determined, and the selectivity index (SI) was calculated from the ratio IC(50)(rat DHFR)/IC(50)(Pc, Tg, or Ma DHFR). The IC(50) values of the 2'-O-carboxypropyl analogue (10), with SI values in parentheses, were 1.1 nM (1300) against Pc DHFR, 9.9 nM (120) against Tg DHFR, and 2.0 nM (600) against Ma DHFR. The corresponding values for the 2'-O-(4 carboxybenzyloxy) analogue (12) were 1.0 nM (560), 22 nM (21), and 0.75 nM (630). By comparison, the IC(50) and SI values for TMP were Pc, 13 000 nM (14); Tg, 2800 nM (65); and Ma, 300 nM (610). For the prototypical potent but nonselective inhibitors PTX and TMX, respectively, these values were Pc, 13 nM (0.26) and 47 nM (0.17); Tg, 4.3 nM (0.76) and 16 nM (0.50); Ma, 0.61 nM (5.4) and 1.5 nM (5.3). Thus 10 and 12 met the criterion for DHFR inhibitors that combine the high selectivity of TMP with the high potency of PTX and TMX. PMID- 15115392 TI - Non-peptidic small-molecule inhibitors of the single-chain hepatitis C virus NS3 protease/NS4A cofactor complex discovered by structure-based NMR screening. AB - NMR-based screening of a customized fragment library identified 16 small-molecule hits that bind weakly (K(D) approximately 100 microM to 10 mM) to substrate binding sites of the NS4A-bound NS3 protease of the hepatitis C virus (HCV). Analogues for five classes of NMR hits were evaluated by a combination of NMR and biochemical data yielding SAR and, in most cases, optimized hits with improved potencies (K(D) approximately K(I) approximately 40 microM to 1 mM). NMR chemical shift perturbation data were used to establish the binding location and orientation of the active site directed scaffolds in these five analogue series. Two of these scaffolds, which bind the enzyme at the proximal S1-S3 and S2' substrate binding sites, were linked together producing competitive inhibitors of the HCV NS3 protease with potencies in the micromolar range. This example illustrates that the low molecular weight scaffolds discovered from structure based NMR screening can be optimized with focused structure-guided chemistry to produce potent nonpeptidic small-molecule inhibitors of the HCV NS3 protease. PMID- 15115393 TI - Conformational analysis of drug-like molecules bound to proteins: an extensive study of ligand reorganization upon binding. AB - This paper describes a large-scale study on the nature and the energetics of the conformational changes drug-like molecules experience upon binding. Ligand strain energies and conformational reorganization were analyzed with different computational methods on 150 crystal structures of pharmaceutically relevant protein-ligand complexes. The common knowledge that ligands rarely bind in their lowest calculated energy conformation was confirmed. Additionally, we found that over 60% of the ligands do not bind in a local minimum conformation. While approximately 60% of the ligands were calculated to bind with strain energies lower than 5 kcal/mol, strain energies over 9 kcal/mol were calculated in at least 10% of the cases regardless of the method used. A clear correlation was found between acceptable strain energy and ligand flexibility, while there was no correlation between strain energy and binding affinity, thus indicating that expensive conformational rearrangements can be tolerated in some cases without overly penalizing the tightness of binding. On the basis of the trends observed, thresholds for the acceptable strain energies of bioactive conformations were defined with consideration of the impact of ligand flexibility. An analysis of the degree of folding of the bound ligands confirmed the general tendency of small molecules to bind in an extended conformation. The results suggest that the unfolding of hydrophobic ligands during binding, which exposes hydrophobic surfaces to contact with protein residues, could be one of the factors accounting for high reorganization energies. Finally, different methods for conformational analysis were evaluated, and guidelines were defined to maximize the prevalence of bioactive conformations in computationally generated ensembles. PMID- 15115394 TI - Peptide-based inhibitors of the hepatitis C virus NS3 protease: structure activity relationship at the C-terminal position. AB - The structure-activity relationship at the C-terminal position of peptide-based inhibitors of the hepatitis C virus NS3 protease is presented. The observation that the N-terminal cleavage product (DDIVPC-OH) of a substrate derived from the NS5A/5B cleavage site was a competitive inhibitor of the NS3 protease was previously described. The chemically unstable cysteine residue found at the P1 position of these peptide-based inhibitors could be replaced with a norvaline residue, at the expense of a substantial drop in the enzymatic activity. The fact that an aminocyclopropane carboxylic acid (ACCA) residue at the P1 position of a tetrapeptide such as 1 led to a significant gain in the inhibitory enzymatic activity, as compared to the corresponding norvaline derivative 2, prompted a systematic study of substituent effects on the three-membered ring. We report herein that the incorporation of a vinyl group with the proper configuration onto this small cycle produced inhibitors of the protease with much improved in vitro potency. The vinyl-ACCA is the first reported carboxylic acid containing a P1 residue that produced NS3 protease inhibitors that are significantly more active than inhibitors containing a cysteine at the same position. PMID- 15115395 TI - Structure-function relationships of multidrug resistance P-glycoprotein. AB - The direct structure-function relationships of P-glycoprotein (P-gp) are presently unknown. In this paper two P-gp models are described: a homology model based on the Escherichia coli MsbA lipid transporter and a model based on the cross-linking results of Loo and Clarke. The pharmacophore pattern for the H-site (Hoechst 33342) is derived and binding sites on the transmembrane domains TM5 and TM11 are identified. Binding sites of rhodamines are also proposed on TM6 and TM12 in accordance with the published data. Location of the binding sites is opposite in both models, suggesting that TMs undergo rotation exposing the substrate bound from the membrane to the pore. It has been concluded that the models derived represent two different functional states of P-gp corresponding to nucleotide-free and nucleotide-bound P-gp. A qualitative correspondence to the P gp crystallographic structure at 20 A resolution is found. A hypothesis is proposed about rearrangement of TMs upon state transition. PMID- 15115396 TI - General model for estimation of the inhibition of protein kinases using Monte Carlo simulations. AB - Monte Carlo statistical mechanics simulations were used in combination with the extended linear response (ELR) approach to develop a model to predict the activities of kinase inhibitors. One hundred forty eight inhibitors of three protein kinases, cyclin-dependent kinase 2 (CDK2), lymphocyte-specific kinase (Lck), and p38 mitogen-activated protein kinase were considered. The inhibitor sets for the individual kinases were analyzed first, and ELR models using only three descriptors were obtained with correlation coefficients, r(2), of 0.7-0.8. Models for each pair of kinases were then developed and used to predict the activities of the inhibitors for the remaining kinase with resultant q(2) values of 0.71 (CDK2), 0.70 (Lck), and 0.54 (p38). Finally, the three datasets were combined to yield a general ELR model for kinase inhibition; with just three physically reasonable descriptors, EXX, DeltaHB(total), and DeltaSASA, the r(2) and leave-one-out q(2) are 0.69 and 0.67. The optimization of the model was confirmed using a genetic algorithm. The descriptors reflect the structural requirements for strong inhibition: good steric and electrostatic complementarities between inhibitor and protein, limited loss of hydrogen bonds for the inhibitor upon binding, and increased burial of surface area of the inhibitor. PMID- 15115397 TI - Roles of conformational and positional adaptability in structure-based design of TMC125-R165335 (etravirine) and related non-nucleoside reverse transcriptase inhibitors that are highly potent and effective against wild-type and drug resistant HIV-1 variants. AB - Anti-AIDS drug candidate and non-nucleoside reverse transcriptase inhibitor (NNRTI) TMC125-R165335 (etravirine) caused an initial drop in viral load similar to that observed with a five-drug combination in naive patients and retains potency in patients infected with NNRTI-resistant HIV-1 variants. TMC125-R165335 and related anti-AIDS drug candidates can bind the enzyme RT in multiple conformations and thereby escape the effects of drug-resistance mutations. Structural studies showed that this inhibitor and other diarylpyrimidine (DAPY) analogues can adapt to changes in the NNRTI-binding pocket in several ways: (1). DAPY analogues can bind in at least two conformationally distinct modes; (2). within a given binding mode, torsional flexibility ("wiggling") of DAPY analogues permits access to numerous conformational variants; and (3). the compact design of the DAPY analogues permits significant repositioning and reorientation (translation and rotation) within the pocket ("jiggling"). Such adaptations appear to be critical for potency against wild-type and a wide range of drug resistant mutant HIV-1 RTs. Exploitation of favorable components of inhibitor conformational flexibility (such as torsional flexibility about strategically located chemical bonds) can be a powerful drug design concept, especially for designing drugs that will be effective against rapidly mutating targets. PMID- 15115398 TI - Rational design and synthesis of novel dimeric diketoacid-containing inhibitors of HIV-1 integrase: implication for binding to two metal ions on the active site of integrase. AB - Discovery of diketoacid-containing compounds as HIV-1 integrase (IN) inhibitors played a major role in validating this enzyme as an important target for the development of therapeutics against HIV infection. In fact, S-1360, the first clinically used IN inhibitor containing a triazole ring as a bioisostere of a carboxylic acid moiety belongs to this class of compounds. To understand the role of divalent metal-chelating in the inhibition of IN (J. Med. Chem. 2002, 45, 5661 5670), we designed and synthesized a series of novel dimeric diketo-containing compounds with the notion that such dimeric compounds may simultaneously bind to two divalent metal ions on the active site of IN. We rationalized that the two diketo subunits separated by uniquely designed linkers can potentially chelate two metal ions that are either provided from one IN active site or two active sites juxtaposed together in a higher order tetramer. Herein, we show that all the new compounds are highly potent against purified IN with varied selectivity for strand transfer, and that some of the analogues exert potent inhibition of the cytopathic effect of HIV-1 in infected CEM cells. This study represents the first attempt to rationally target two divalent metal ions on the active site of IN and may have potential implications for the design of second generation diketoacid-containing class of inhibitors. PMID- 15115399 TI - Design, synthesis, structural studies, biological evaluation, and computational simulations of novel potent AT(1) angiotensin II receptor antagonists based on the 4-phenylquinoline structure. AB - Novel AT(1) receptor antagonists bearing substituted 4-phenylquinoline moieties instead of the classical biphenyl fragment were designed and synthesized as the first step of an investigation devoted to the development of new antihypertensive agents and to the understanding of the molecular basis of their pharmacodynamic and pharmacokinetic properties. The newly synthesized compounds were tested for their potential ability to displace [(125)I]Sar(1),Ile(8)-Ang II specifically bound to AT(1) receptor in rat hepatic membranes. These AT(1) receptor binding studies revealed nanomolar affinity in several of the compounds under study. The most potent ligands 4b,t were found to be equipotent with losartan and possessed either a 3-tetrazolylquinoline or a 2-amino-3-quinolinecarboxylic moiety, respectively. Moreover, some selected compounds were evaluated for antagonism of Ang II-induced contraction in rabbit aortic strips, and the most potent compounds in the binding test 4b,t were slightly more potent than losartan in inhibiting Ang II-induced contraction. Finally, the most relevant structure-affinity relationship data were rationalized by means of computational studies performed on the isolated ligands as well as by computational simulations on the ligands complexed with a theoretical AT(1) receptor model. PMID- 15115400 TI - Synthesis of novel potent dipeptidyl peptidase IV inhibitors with enhanced chemical stability: interplay between the N-terminal amino acid alkyl side chain and the cyclopropyl group of alpha-aminoacyl-l-cis-4,5-methanoprolinenitrile based inhibitors. AB - A series of methanoprolinenitrile-containing dipeptide mimetics were synthesized and assayed as inhibitors of the N-terminal sequence-specific serine protease dipeptidyl peptidase IV (DPP-IV). The catalytic action of DPP-IV is the principle means of degradation of glucagon-like peptide-1, a key mediator of glucose stimulated insulin secretion, and DPP-IV inhibition shows clinical benefit as a novel mechanism for treatment of type 2 diabetes. However, many of the reversible inhibitors to date suffer from chemical instability stemming from an amine to nitrile intramolecular cyclization. Installation of a cyclopropyl moiety at either the 3,4- or 4,5-position of traditional 2-cyanopyrrolidide proline mimetics led to compounds with potent inhibitory activity against the enzyme. Additionally, cis-4,5-methanoprolinenitriles with beta-branching in the N terminal amino acid provided enhanced chemical stability and high inhibitory potency. This class of inhibitors also exhibited the ability to suppress prandial glucose elevations after an oral glucose challenge in male Zucker rats. PMID- 15115401 TI - Oral bioavailability of a new class of micro-opioid receptor agonists containing 3,6-bis[Dmt-NH(CH(2))(n)]-2(1H)-pyrazinone with central-mediated analgesia. AB - The inability of opioid peptides to be transported through epithelial membranes in the gastrointestinal tract and pass the blood-brain barrier limits their effectiveness for oral application in an antinociceptive treatment regime. To overcome this limitation, we enhanced the hydrophobicity while maintaining the aqueous solubility properties in a class of opioid-mimetic substances by inclusion of two identical N-termini consisting of Dmt (2',6'-dimethyl-l tyrosine) coupled to a pyrazinone ring platform by means of alkyl chains to yield the class of 3,6-bis[Dmt-NH-(CH(2))(n)]-2(1H)-pyrazinones. These compounds displayed high micro-opioid receptor affinity (K(i)micro = 0.042-0.115 nM) and selectivity (K(i)delta/K(i)micro = 204-307) and functional micro-opioid receptor agonism (guinea-pig ileum, IC(50) = 1.3-1.9 nM) with little or undetectable bioactivity toward delta-opioid receptors (mouse vas deferens) and produced analgesia in mice in a naloxone reversible manner when administered centrally (intracerebroventricular, i.c.v.) or systemically (subcutaneously and orally). Furthermore, the most potent compound, 3,6-bis(3'-Dmt-aminopropyl)-5-methyl-2(1H) pyrazinone (7'), lacked functional delta-opioid receptor bioactivity and was 50 63-fold and 18-21-fold more active than morphine by icv administration as measured analgesia using tail-flick (spinal involvement) and hot-plate (supraspinal effect) tests, respectively; the compound ranged from 16 to 63% as potent upon systemic injection. These analgesic effects are many times greater than unmodified opioid peptides. The data open new possibilities for the rational design of potential opioid-mimetic drugs that pass through the epithelium of the gastrointestinal tract and the blood-brain barrier to target brain receptors. PMID- 15115402 TI - Cytotoxic alpha-halogenoacrylic derivatives of distamycin A and congeners. AB - The mechanism of action of many antitumor agents involves DNA damage, either by direct binding of the drug to DNA or to DNA-binding proteins. However, most of the DNA-interacting agents have only a limited degree of sequence specificity, which implies that they may hit all the cellular genes. DNA minor groove binders, among which the derivatives of distamycin A play an important role, could provide significant improvement in cancer management, increasing gene specificity, due to high selectivity of interaction with thymine-adenine (TA) rich sequences. We now report and discuss the synthesis, the in vitro and in vivo activities, and some mechanistic features of alpha-halogenoacrylamido derivatives of distamycin A. The final result of this work was the selection of brostallicin 17 (PNU-166196). Brostallicin, presently in phase II clinical trials, shows a broad spectrum of antitumor activity and an apoptotic effect higher than distamycin derivative tallimustine. An important in vitro toxicological feature of brostallicin is the very good ratio between myelotoxicity on human haematopoietic progenitor cells and cytotoxicity on tumor cells, in comparison with clinically tested DNA minor groove binders. A peculiarity of brostallicin is its in vitro reactivity in the DNA alkylation assays only in the presence of glutathione. Moreover brostallicin's antitumor activity, both in in vitro and in vivo tumor models, is higher in the presence of increased levels of glutathione/glutathione-S tranferases. These findings contribute to the definition of brostallicin as a novel anticancer agent that differs from other minor groove binders and alkylating agents for both the profile of activity and the mechanism of action and to classify the alpha-bromoacrylamido derivatives of distamycin as a new class of cytotoxics. Moreover, due to its interaction with glutathione, brostallicin may have a role for the tailored treatment of tumors characterized by constitutive or therapy-induced overexpression of glutathione/glutathione-S tranferase levels. PMID- 15115403 TI - Synthesis and pharmacological evaluation of 3-(3,4-dichlorophenyl)-1-indanamine derivatives as nonselective ligands for biogenic amine transporters. AB - In our efforts toward developing a nonselective ligand that would block the effects of stimulants such as methamphetamine at dopamine (DA), serotonin (5-HT), and norepinephrine (NE) transporters, we synthesized a series of 3-(3,4 dichlorophenyl)-1-indanamine derivatives. Two of the examined higher affinity compounds had a phenolic hydroxyl group enabling preparation of a medium to long chain carboxylic acid ester that might eventually be useful for a long-acting depot formulation. The in vitro data indicated that (-)-(1R,3S)-trans-3-(3,4 dichlorophenyl)-6-hydroxy-N-methyl-1-indanamine ((-)-(1R,3S)-11) displays high affinity binding and potent inhibition of uptake at all three biogenic amine transporters. In vivo microdialysis experiments demonstrated that intravenous administration of (-)-(1R,3S)-11 to rats elevated extracellular DA and 5-HT in the nucleus accumbens in a dose-dependent manner. Pretreating rats with 0.5 mg/kg (-)-(1R,3S)-11 elevated extracellular DA and 5-HT by approximately 150% and reduced methamphetamine-induced neurotransmitter release by about 50%. Ex vivo autoradiography, however, demonstrated that iv administration of (-)-(1R,3S)-11 produced a dose-dependent, persistent occupation of 5-HT transporter binding sites but not DA transporter sites. PMID- 15115404 TI - Inhibitors of Sir2: evaluation of splitomicin analogues. AB - Splitomicin (1) and 41 analogues were prepared and evaluated in cell-based Sir2 inhibition and toxicity assays and an in vitro Sir2 inhibition assay. Lactone ring or naphthalene (positions 7-9) substituents decrease activity, but other naphthalene substitutions (positions 5 and 6) are well-tolerated. The hydrolytically unstable aromatic lactone is important for activity. Lactone hydrolysis rates were used as a measure of reactivity; hydrolysis rates correlate with inhibitory activity. The most potent Sir2 inhibitors were structurally similar to and had hydrolysis rates similar to 1. PMID- 15115405 TI - Prediction of biological targets using probabilistic neural networks and atom type descriptors. AB - Prediction of biological targets for molecules from their chemical structures is beneficial for generating focused libraries, selecting compounds for screening, and annotating biological activities for those compounds whose activities are unknown. We studied the ability of a probabilistic neural network (PNN), a variant of normalized radial basis function (RBF) neural networks, to predict biological activities for a set of 799 compounds having activities against seven biological targets. The compounds were taken from the MDDR database, and they were carefully selected to comprise distinct biological activities and diverse structures. The structural characteristics of compounds were represented by a set of 24 atom-type descriptors defined by 2D topological chemical structures. The modeling was done in two ways: (1). compounds having one certain activity were discriminated from those not having that activity and (2). all compounds were classified into seven biological classes. In both cases, around 90% of the compounds were correctly classified. Further validation of the modeled PNNs was done with 26 317 compounds having biological activities against various targets except for the seven targets used for modeling, and 67-98% compounds were correctly classified depending upon the targets. A PNN trains much more quickly than widely used neural networks such as a feed-forward neural network with error back-propagation. Calculation of atom-type descriptors is easy even for a large size chemical library. Combination of PNN and atom-type descriptors thus provides a powerful way to predict biological activities from structural information. PMID- 15115406 TI - Significant differences in biological parameters between prodrugs cleavable by carboxypeptidase G2 that generate 3,5-difluoro-phenol and -aniline nitrogen mustards in gene-directed enzyme prodrug therapy systems. AB - Nine new nitrogen mustard compounds derived from 2,6-difluoro-4-hydroxy- (3a-e) and 2,6-difluoro-4-amino- (4a-d) aniline were synthesized as potential prodrugs. They were designed to be activated to their corresponding 3,5-difluorophenol and aniline (4)-nitrogen mustards by the enzyme carboxypeptidase G2 (CPG2) in gene directed enzyme prodrug therapy (GDEPT) models. The compounds were tested for cytotoxicity in the MDA MB-361 breast adenocarcinoma. The cell line was engineered to express stably either CPG2 tethered to the cell surface stCPG2-(Q)3 or beta-galactosidase (beta-Gal) as control. The cytotoxicity differentials were calculated between CPG 2-expressing and -nonexpressing cells and yielded different results for the two series of prodrugs despite their structural similarities. While the phenol compounds are ineffective as prodrugs, their aniline counterparts exhibit outstanding activity in the tumor cell lines expressing CPG2. [3,5-Difluoro-4-[bis(2-chloroethyl)amino]phenyl]carbamoyl-l glutamic acid gave a differential of >227 in MDA MB361 cells as compared with 19 exhibited by 4-[(2-chloroethyl)(2-mesyloxyethyl)amino]benzoyl-l-glutamic acid, 1a, which has been in clinical trials. PMID- 15115407 TI - New arylpiperazine 5-HT(1A) receptor ligands containing the pyrimido[2,1-f]purine fragment: synthesis, in vitro, and in vivo pharmacological evaluation. AB - New 1H,3H-pyrimido[2,1-f]purine-2,4-dione derivatives of arylpiperazine (11-22) were prepared and evaluated in vitro for their affinity for 5-HT(1A), 5-HT(2A), alpha(1), and D(2) receptors. The tested compounds showed high affinity for 5 HT(1A) and alpha(1) receptors (K(i) = 1.1-87 and 10-62 nM, respectively) and moderate to low affinity for 5-HT(2A) (K(i) = 56-881 nM) and D(2) receptors (K(i) = 94-1245 nM). Compounds 14, 15, 18, 19, and 21, mostly 3'-chlorophenylpiperazine derivatives, can be classified as mixed 5-HT(1A)/5-HT(2A)/alpha(1) ligands. Compound 13, which showed the highest 5-HT(1A) receptor affinity (K(i) = 1.1 nM), was 50-fold selective in relation to alpha(1) adrenoceptors and at least 250-fold over 5-HT(2A) and D(2) sites. On the basis of in vivo functional tests, 8 phenylpiperazinoethylamino (11), 8-(2'-methoxyphenylpiperazino)ethylamino (13), and 8-phenylpiperazinopropylamino (14) derivatives of 1,3-dimethyl-1H,3H pyrimido[2,1-f]purine-2,4-dione were identified as potent pre- and postsynaptic 5 HT(1A) receptor antagonists. 1,3-Dimethyl-7-bromo-8-(phenylpiperazinopropylamino) 1H,3H-pyrimido[2,1-f]purine-2,4-dione (20) behaved like an agonist of presynaptic and as a partial agonist of postsynaptic 5-HT(1A) receptors and resembled ipsapirone in terms of functional intrinsic activity. It revealed marked anxiolytic-like activity in the Vogel test in rats, comparable to that of the reference drug diazepam, and exhibited antidepressant-like activity in the Porsolt test in rats. The sedative effect of 20, evaluated in the open field test in rats, appeared at doses twice as high as those inducing a minimal anxiolytic like effect and was similar to the effects of diazepam. PMID- 15115408 TI - A new class of nonpeptide bradykinin B(2) receptor ligand, incorporating a 4 aminoquinoline framework. Identification of a key pharmacophore to determine species difference and agonist/antagonist profile. AB - Introduction of various aliphatic amino groups at the 4-position of the quinoline moiety of our nonpeptide bradykinin (BK) B(2) receptor antagonists afforded highly potent ligands for human B(2) receptor with various affinities for guinea pig B(2) receptor, indicating remarkable species difference. A representative 4 dimethyamino derivative 40a exhibited subnanomolar and nanomolar binding affinities for human and guinea pig B(2) receptors, respectively, and significantly inhibited BK-induced bronchoconstriction in guinea pigs at 10 microg/kg by intravenous administration. Further chemical modification led us to discover unique partial agonists for the human B(2) receptor that increase inositol phosphates (IPs) production by themselves in Chinese hamster ovary (CHO) cells expressing the cloned human B(2) receptor. Although their potency and efficacy were much lower than those of BK, we identified them as screening leads for nonpeptide B(2) agonists. In these studies it was revealed the 4-substituent of the quinoline moiety is the key pharmacophore to determine species difference and agonist/antagonist profiles. PMID- 15115409 TI - 4-(omega-(alkyloxy)alkyl)-1H-imidazole derivatives as histamine H(3) receptor antagonists/agonists. AB - In an effort to develop new histamine H(3) receptor antagonists usable as pharmacological tools we present here novel unsymmetrical ether derivatives. Etherification of different omega-(1H-imidazol-4-yl)alkyl scaffolds led to compounds containing alkyl chains of increasing lengths either with or without unsaturated termini, cycloalkyl or arylalkyl moieties, or additional heteroatoms. When investigated in an in vitro assay on rat synaptosomes, the majority of compounds displayed potencies in the low nanomolar concentration range at the H(3) receptor, e.g., 4-(3-(3-cyclopentylpropyloxy)propyl)-1H-imidazole (27, K(i) = 7 nM). FUB 465, 4-(3-(ethoxy)propyl)-1H-imidazole (14), a useful tool for the characterization of constitutive activity of H(3) receptors in vivo in rodents, proved to be of high oral in vivo potency in mice (ED(50) = 0.26 mg/kg). Further, the influence of chosen compounds on specific [(35)S]GTPgammaS binding was assayed on HEK293 cell membranes expressing the human histamine H(3) receptor revealing partial agonism of the compounds in this particular model. These distinct responses are further hints for "protean agonism" in this class of compounds. Additionally, selected compounds were functionally investigated in vitro on isolated organs of the guinea-pig at H(3), H(1), and H(2) receptors. PMID- 15115410 TI - New 1,4-dihydropyridines endowed with NO-donor and calcium channel agonist properties. AB - A new series of calcium channel agonists structurally related to Bay K8644, containing NO donor furoxans and the related furazans unable to release NO, is described. The racemic mixtures were studied for their action on L-type Ca(2+) channels expressed in cultured rat insulinoma RINm5F cells. All the products proved to be potent calcium channel agonists. All the racemic mixtures, with the only exception of the carbamoyl derivatives 9, 12 endowed with scanty solubility, were separated by chiral chromatography into the corresponding enantiomers; the (+) enantiomers were found to be potent agonists while the (-) ones were feeble antagonists. The racemic mixtures were also assessed for their positive inotropic activity on electrically stimulated rat papillary muscle and for their ability to increase Ca(2+) entry into the vascular smooth muscle of rat aorta strips. The cyanofuroxan 8 proved to be an interesting product with dual Ca(2+)-dependent positive inotropic and NO-dependent vasodilating activity. PMID- 15115411 TI - Fluoroartemisinin: trifluoromethyl analogues of artemether and artesunate. AB - The synthesis of a series of C-10 trifluoromethyl ethers of artemisinin has been achieved from key bromide 8, itself carried out in two steps from artemisinin. The substitution of 8 with methanol, ethanol, or succinic acid allowed the access of C-10 CF(3) analogues of beta-artemether, beta-arteether, or artesunate, respectively, in good yields (up to 89%). The presence of the CF(3) group at C-10 of artemisinin clearly increased the chemical stability under simulated stomach acid conditions. For example, the CF(3) analogue of arteether was found to be around 45 times more stable than arteether itself. The influence of the CF(3) moiety on biological activity was also highlighted. CF(3) analogues of artemether and arteether exhibited a high in vivo antimalarial activity on mice infected with Plasmodium berghei NK173, with a complete clearance of the parasitemia during the entire observation period (25 days). PMID- 15115412 TI - Parallel solution-phase synthesis of conformationally restricted congeners of pentamidine and evaluation of their antiplasmodial activities. AB - Conformationally restricted bisbenzamidines and related congeners have been synthesized and evaluated for activity against two Plasmodium falciparum strains. The most active compounds, bisbenzamidines linked by a 1,4-piperazinediyl core, had IC(50) values between 3 and 18 nM against both chloroquine-susceptible and resistant parasites and IC(50) values for cytotoxicity greater than 5 microM, using the A549 human lung epithelial cell line. DNA binding affinity, as estimated by DeltaT(m), did not correlate with either antiparasite effects or cytotoxicity. Each of the active bisbenzamidines interfered with the formation of hemozoin in cell-free systems. PMID- 15115413 TI - [1-(3,5-difluoro-4-hydroxyphenyl)-1H-pyrrol-3-yl]phenylmethanone as a bioisostere of a carboxylic acid aldose reductase inhibitor. AB - [1-(3,5-Difluoro-4-hydroxyphenyl)-1H-pyrrol-3-yl]phenylmethanone (6) was synthesized as a putative bioisostere of the known aldose reductase (AR) inhibitor (3-benzoylpyrrol-1-yl)acetic acid (I). It was found that 6 is approximately 5 times more potent as an in vitro inhibitor of AR than I, with an IC(50) value in the submicromolar range. Furthermore, 6 showed considerable activity in an in vitro experimental glycation model of diabetes mellitus. Our results support the notion that 6 might become a useful lead structure. PMID- 15115414 TI - Synthesis and use of novel ether phospholipid enantiomers to probe the molecular basis of the antitumor Effects of alkyllysophospholipids: correlation of differential activation of c-Jun NH(2)-terminal protein kinase with antiproliferative effects in neuronal tumor cells. AB - The enantiomers of a novel unsaturated phosphonocholine antitumor ether lipid were synthesized and found to have differential antiproliferative effects against epithelial cancer cell lines. The basis of the enantioselective effects on the cells was investigated in SK-N-MC and SK-N-SH neuroblastoma tumor cells. Our results indicate that the enantioselective antiproliferative potency arises primarily from the activation of the JNK signaling pathway by the ether lipids. PMID- 15115417 TI - Prevention of recurrent thrombosis in the antiphospholipid antibody syndrome: how long and how high with oral anticoagulant therapy? PMID- 15115418 TI - To exercise or not to exercise in chronic fatigue syndrome? No longer a question. PMID- 15115419 TI - Estimating disease likelihood: a case of rubbery figures. PMID- 15115420 TI - Modernisation or reform? The NHS approach. PMID- 15115421 TI - Randomised controlled trial of graded exercise in chronic fatigue syndrome. AB - OBJECTIVE: To investigate whether 12 weeks of graded exercise with pacing would improve specific physiological, psychological and cognitive functions in people with chronic fatigue syndrome (CFS). DESIGN: Randomised controlled trial. SETTING: Human performance laboratory at the University of Western Australia. PARTICIPANTS: 61 patients aged between 16 and 74 years diagnosed with CFS. INTERVENTIONS: Either graded exercise with pacing (32 patients) or relaxation/flexibility therapy (29 patients) performed twice a day over 12 weeks. MAIN OUTCOME MEASURES: Changes in any of the physiological, psychological or cognitive variables assessed. RESULTS: Following the graded exercise intervention, scores were improved for resting systolic blood pressure (P = 0.018), work capacity (W.kg(-1)) (P = 0.019), net blood lactate production (P = 0.036), depression (P = 0.027) and performance on a modified Stroop Colour Word test (P = 0.029). Rating of perceived exertion scores, associated with an exercise test, was lower after graded exercise (P = 0.013). No such changes were observed in the relaxation/flexibility condition, which served as an attention placebo control. CONCLUSIONS: Graded exercise was associated with improvements in physical work capacity, as well as in specific psychological and cognitive variables. Improvements may be associated with the abandonment of avoidance behaviours. PMID- 15115422 TI - Generating pre-test probabilities: a neglected area in clinical decision making. AB - OBJECTIVE: To assess the accuracy and variability of clinicians' estimates of pre test probability for three common clinical scenarios. DESIGN: Postal questionnaire survey conducted between April and October 2001 eliciting pre-test probability estimates from scenarios for risk of ischaemic heart disease (IHD), deep vein thrombosis (DVT), and stroke. PARTICIPANTS AND SETTING: Physicians and general practitioners randomly drawn from College membership lists for New South Wales and north-west England. MAIN OUTCOME MEASURES: Agreement with the "correct" estimate (being within 10, 20, 30, or > 30 percentage points of the "correct" estimate derived from validated clinical-decision rules); variability in estimates (median and interquartile ranges of estimates); and association of demographic, practice, or educational factors with accuracy (using linear regression analysis). RESULTS: 819 doctors participated: 310 GPs and 288 physicians in Australia, and 106 GPs and 115 physicians in the UK. Accuracy varied from about 55% of respondents being within 20% of the "correct" risk estimate for the IHD and stroke scenarios to 6.7% for the DVT scenario. Although median estimates varied between the UK and Australian participants, both were similar in accuracy and showed a similarly wide spread of estimates. No demographic, practice, or educational variables substantially predicted accuracy. CONCLUSIONS: Experienced clinicians, in response to the same clinical scenarios, gave a wide range of estimates for pre-test probability. The development and dissemination of clinical decision rules is needed to support decision making by practising clinicians. PMID- 15115423 TI - Impact of an electronic antibiotic advice and approval system on antibiotic prescribing in an Australian teaching hospital. AB - The impact of a computer-based infectious diseases electronic antibiotic advice and approval system ("IDEA(3)S") was assessed as an alternative to a labour intensive, phone-based approval system. IDEA(3)S-based approvals replaced 48% of all approvals for the most frequently requested antimicrobial agents (ceftriaxone/cefotaxime, vancomycin) and were associated with stable overall rates of antimicrobial use. Antibiotic prescribing for community-acquired pneumonia was 76% concordant with IDEA(3)S recommendations, and clinical acceptance of IDEA(3)S was excellent. Successful implementation required a coordinated, evidence-based approach between clinicians, pharmacists and hospital administration, together with ongoing staff education and feedback of results. IDEA(3)S is a useful new adjunct to routine clinician consultation to support appropriate antibiotic prescribing for a number of common indications in hospitals. PMID- 15115424 TI - Increase in type 2 diabetes in children and adolescents in Western Australia. AB - OBJECTIVES: To document diagnosis rates of type 2 diabetes mellitus in children and adolescents in Western Australia over the past 12 years, the clinical characteristics of these patients and any comorbidities. DESIGN: Review of a prospectively recorded diabetes database. SETTING: Tertiary paediatric referral centre (the only such centre in WA). PATIENTS: All children and adolescents aged < 17 years diagnosed with type 2 diabetes between 1990 and 2002 and managed by Princess Margaret Hospital Diabetes Unit. MAIN OUTCOME MEASURES: Anthropometric and demographic data; glycohaemoglobin (HbA(1c)) level; blood pressure; lipid levels; presence of acanthosis nigricans. RESULTS: 43 patients (15 males and 28 females) were diagnosed with type 2 diabetes. Age (SD) at diagnosis was 13.6 (1.8) years. The rate of diagnosis has been progressively increasing (average annual increase in the unadjusted overall rates of type 2 diabetes was 27%). Twenty-three patients (53%) were of Indigenous origin and 18 (42%) resided in rural areas. The mean (SD) HbA(1c) level at diagnosis was 10.0% (3.2%). Seventy two per cent of patients had acanthosis nigricans, 59% had hypertension, and 24% had hyperlipidaemia. CONCLUSIONS: There has been an increase in the diagnosis rate of type 2 diabetes in children and adolescents in WA. Comorbidities are frequent. PMID- 15115427 TI - Does high-impact exercise in the prepubertal period have an osteogenic effect in females? PMID- 15115425 TI - Metformin therapy and diabetes in pregnancy. AB - No adverse pregnancy outcomes with metformin use have been reported, except in one unmatched study. Otherwise, the studies are small and non-randomised, with the exception of one prospective, randomised controlled trial, currently under way, comparing metformin with insulin in women with gestational diabetes mellitus (the MiG trial). No long-term follow-up data for offspring of mothers receiving metformin have been published. Any woman with diabetes should be as close to euglycaemia as possible before pregnancy. In some circumstances (eg, severe insulin resistance), metformin therapy during pregnancy may be warranted. When metformin treatment is being considered, the individual risks and benefits need to be discussed with the patient so that an appropriate decision can be reached. PMID- 15115428 TI - Subgroup analysis: application to individual patient decisions. PMID- 15115429 TI - Successful reintroduction of statin therapy after myositis: was there another cause? PMID- 15115430 TI - 10: Management of obesity. AB - Obesity has reached epidemic proportions in Australia, with 67.5% of men, 52.1% of women and 19%-23% of children and adolescents being overweight or obese. Genetically predisposed individuals are especially vulnerable to developing obesity in the highly obesogenic environment of 21st century Australia. Obesity causes or contributes to many comorbidities, including type 2 diabetes, hypertension, dyslipidaemia, sleep apnoea, non-alcoholic steatohepatitis, orthopaedic problems and polycystic ovary syndrome. Management in the individual requires their complete co-operation and should be tailored to individual needs and complications. Management of obesity in children should consider the family context and involve the parents. All treatment strategies must involve lifestyle modification, with a reduction of energy intake and an increase in physical activity. Some patients may also require the assistance of drug therapy or bariatric surgery. PMID- 15115431 TI - Murine typhus: the first reported case from Victoria. PMID- 15115432 TI - Sublingual glyceryl trinitrate as prehospital treatment for hypertension in Irukandji syndrome. PMID- 15115433 TI - Travel insurance and medical evacuation. PMID- 15115434 TI - Are the Australian guidelines asking too much of the Pneumonia Severity Index (PSI)? PMID- 15115435 TI - El Nino Southern Oscillation and the transmission of hepatitis A virus in Australia. PMID- 15115436 TI - Deletion of Ser-171 causes inactivation, proteasome-mediated degradation and complete deficiency of human transaldolase. AB - Homozygous deletion of three nucleotides coding for Ser-171 (S171) of TAL-H (human transaldolase) has been identified in a female patient with liver cirrhosis. Accumulation of sedoheptulose 7-phosphate raised the possibility of TAL (transaldolase) deficiency in this patient. In the present study, we show that the mutant TAL-H gene was effectively transcribed into mRNA, whereas no expression of the TALDeltaS171 protein or enzyme activity was detected in TALDeltaS171 fibroblasts or lymphoblasts. Unlike wild-type TAL-H-GST fusion protein (where GST stands for glutathione S-transferase), TALDeltaS171-GST was solubilized only in the presence of detergents, suggesting that deletion of Ser 171 caused conformational changes. Recombinant TALDeltaS171 had no enzymic activity. TALDeltaS171 was effectively translated in vitro using rabbit reticulocyte lysates, indicating that the absence of TAL-H protein in TALDeltaS171 fibroblasts and lymphoblasts may be attributed primarily to rapid degradation. Treatment with cell-permeable proteasome inhibitors led to the accumulation of TALDeltaS171 in whole cell lysates and cytosolic extracts of patient lymphoblasts, suggesting that deletion of Ser-171 led to rapid degradation by the proteasome. Although the TALDeltaS171 protein became readily detectable in proteasome inhibitor-treated cells, it displayed no appreciable enzymic activity. The results suggest that deletion of Ser-171 leads to inactivation and proteasome-mediated degradation of TAL-H. Since TAL-H is a regulator of apoptosis signal processing, complete deficiency of TAL-H may be relevant for the pathogenesis of liver cirrhosis. PMID- 15115438 TI - Identification of the first archaeal Type 1 RNase H gene from Halobacterium sp. NRC-1: archaeal RNase HI can cleave an RNA-DNA junction. AB - All the archaeal genomes sequenced to date contain a single Type 2 RNase H gene. We found that the genome of a halophilic archaeon, Halobacterium sp. NRC-1, contains an open reading frame with similarity to Type 1 RNase H. The protein encoded by the Vng0255c gene, possessed amino acid sequence identities of 33% with Escherichia coli RNase HI and 34% with a Bacillus subtilis RNase HI homologue. The B. subtilis RNase HI homologue, however, lacks amino acid sequences corresponding to a basic protrusion region of the E. coli RNase HI, and the Vng0255c has the similar deletion. As this deletion apparently conferred a complete loss of RNase H activity on the B. subtilis RNase HI homologue protein, the Vng0255c product was expected to exhibit no RNase H activity. However, the purified recombinant Vng0255c protein specifically cleaved an RNA strand of the RNA/DNA hybrid in vitro, and when the Vng0255c gene was expressed in an E. coli strain MIC2067 it could suppress the temperature-sensitive growth defect associated with the loss of RNase H enzymes of this strain. These results in vitro and in vivo strongly indicate that the Halobacterium Vng0255c is the first archaeal Type 1 RNase H. This enzyme, unlike other Type 1 RNases H, was able to cleave an Okazaki fragment-like substrate at the junction between the 3'-side of ribonucleotide and 5'-side of deoxyribonucleotide. It is likely that the archaeal Type 1 RNase H plays a role in the removal of the last ribonucleotide of the RNA primer from the Okazaki fragment during DNA replication. PMID- 15115437 TI - Regulation of Cyp2a5 transcription in mouse primary hepatocytes: roles of hepatocyte nuclear factor 4 and nuclear factor I. AB - The cytochrome P4502a5 (Cyp2a5) gene is expressed principally in liver and olfactory mucosa. In the present study, the transcriptional mechanisms of hepatocyte-specific expression of Cyp2a5 were studied in mouse primary hepatocytes. The Cyp2a5 5'-flanking region -3033 to +10 was cloned in front of a luciferase reporter gene and transfected into hepatocytes. Deletion analysis revealed two major activating promoter regions localized at proximal 271 bp and at a more distal area from -3033 to -2014 bp. The proximal activation region was characterized further by DNase I footprinting, and a single clear footprint was detected in the studied area centred over a sequence similar to the NF-I (nuclear factor I)-binding site. The binding of NF-I was confirmed using an EMSA (electrophoretic mobility-shift assay). A putative HNF-4 (hepatocyte nuclear factor 4)-binding site was localized at the proximal promoter by computer analysis of the sequence, and HNF-4alpha was shown to interact with the site using an EMSA. The functional significance of HNF-4 and NF-I binding to the Cyp2a5 promoter was evaluated by site-directed mutagenesis of the binding motifs in reporter constructs. Both mutations strongly decreased transcriptional activation by the Cyp2a5 promoter in primary hepatocytes, and double mutation almost completely abolished transcriptional activity. Also, the functionality of the distal activation region was found to be dependent on the intact HNF-4 and NF I sites at the proximal promoter. In conclusion, these results indicate that HNF 4 and NF-I play major roles in the constitutive regulation of hepatic expression of Cyp2a5. PMID- 15115439 TI - Cloning and expression of an inhibitor of microbial metalloproteinases from insects contributing to innate immunity. AB - The first IMPI (inhibitor of metalloproteinases from insects) was identified in the greater wax moth, Galleria mellonella [Wedde, Weise, Kopacek, Franke and Vilcinskas (1998) Eur. J. Biochem. 255, 535-543]. Here we report cloning and expression of a cDNA coding for this IMPI. The IMPI mRNA was identified among the induced transcripts from a subtractive and suppressive PCR analysis after bacterial challenge of G. mellonella larvae. Induced expression of the IMPI during a humoral immune response was confirmed by real-time PCR, which documented up to 500 times higher amounts of IMPI mRNA in immunized larvae in comparison with untreated ones. The IMPI sequence shares no similarity with those of tissue inhibitors of metalloproteinases or other natural inhibitors of metalloproteinases, and the recombinant IMPI specifically inhibits thermolysin like metalloproteinases, but not matrix metalloproteinases. These results support the hypothesis that the IMPI represents a novel type of immune-related protein which is induced and processed during the G. mellonella humoral immune response to inactivate pathogen-associated thermolysin-like metalloproteinases. PMID- 15115440 TI - The use of ciclosporin in psoriasis: a clinical review. AB - Psoriasis is a chronic condition that shows variability in phenotype and severity. The disease can seriously compromise patients' quality of life, regardless of disease extent. Systemic treatment is indicated when lesional burden is extensive and/or frequently relapsing, and when quality of life is severely altered. Furthermore, surveys have indicated that patients are dissatisfied with their current topical or phototherapy. The efficacy of ciclosporin in the treatment of psoriasis is well established. However, widespread use of this drug has been limited by concerns over adverse effects, such as renal impairment, hypertension and the potential risk of malignancy. Data from many clinical trials designed to examine the efficacy and safety of long term continuous and intermittent short-course (< 12 weeks) therapy are now available. Information from these studies has aided dermatologists in developing treatment guidelines. Intermittent short-course therapy is well tolerated, safe, and highly effective in sustaining disease control and promoting quality of life. Long-term continuous ciclosporin therapy may be useful in some patients with refractory psoriasis. If treatment guidelines are followed, the risk of nephrotoxicity and hypertension is low. Ciclosporin therapy is associated with an increased risk of non-melanoma skin cancer (mainly squamous cell carcinoma) when patients have been previously exposed to psoralen-ultraviolet A (PUVA). The incidence of non-skin malignancy shows no significant difference to that observed in the general population. PMID- 15115441 TI - Ciclosporin in psoriasis clinical practice: an international consensus statement. AB - The main recommendations for the use of ciclosporin in the management of psoriasis are: (i) intermittent short courses (average of 12 weeks duration) of ciclosporin are preferable; (ii) ciclosporin should be given in the dose range 2.5-5.0 mg kg(-1) day(-1) (doses greater than 5.0 mg kg(-1) day(-1) should only be given in exceptional circumstances); (iii) treatment regimens should be tailored to the needs of each patient; (iv) selection of patients should take into account psychosocial disability, as well as clinical extent of disease and failure of previous treatment; (v) each patient's renal function (as measured by serum creatinine) should be thoroughly assessed before and during treatment; (vi) each patient's blood pressure should be carefully monitored before and during treatment; (vii) adherence to treatment guidelines substantially reduces the risk of adverse events; (viii) long-term continuous ciclosporin therapy may be appropriate in a subgroup of patients; however, duration of treatment should be kept below 2 years whenever possible; and (ix) when long-term continuous ciclosporin therapy is necessary, annual evaluation of glomerular filtration rate may be useful to accurately monitor renal function. PMID- 15115444 TI - Prevalence of pseudoexfoliation syndrome in Ethiopian patients scheduled for cataract surgery. AB - PURPOSE: To evaluate the prevalence of pseudoexfoliation syndrome (PEX) among Ethiopian patients with age-related cataract scheduled for surgery. PATIENTS AND METHODS: In a prospective study, 229 consecutive eyes with age-related cataract scheduled for surgery were evaluated for the prevalence of PEX, type of cataract, intraocular pressure (IOP) and glaucoma. RESULTS: The mean age of the 229 patients was 62.1 +/- 10.5 years (range 47-91 years). The prevalence of PEX was 39.3% (90 of 229 eyes). Pseudoexfoliation syndrome was bilateral in 62.2% (56/90) of cases. The mean age of those with PEX (63.7 +/- 10.7 years) was significantly more than that of those without PEX (60.3 +/- 9.2 years) (p = 0.01). The prevalence of PEX significantly increased with age (p = 0.02). Patients with hypermature cataract had significantly more PEX (p < 0.001). Mean IOP was significantly higher in eyes with PEX (18.4 +/- 3.2 mmHg) than in those without PEX (15.8 +/- 3.6 mmHg) (p < 0.001). Significantly more eyes in the PEX group had IOP > 21 mmHg (p < 0.01) and established glaucoma (p < 0.05). An incidence of PEX was found in all major ethnic groups in Ethiopia. There was no significant difference in the prevalence of PEX between the different ethnic groups (p > 0.5). CONCLUSIONS: The prevalence of PEX among Ethiopian patients with cataract is high and occurs at a relatively younger age when compared to previous reports from Europe. Pseudoexfoliation syndrome has been found to be significantly associated with hypermature cataract, higher mean IOP, IOP > 21 mmHg and glaucoma. Population-based studies are recommended to evaluate the prevalence of PEX in the general population and its association with cataract and glaucoma. PMID- 15115445 TI - Exfoliation syndrome in Estonian patients scheduled for cataract surgery. AB - PURPOSE: To examine the frequency of exfoliation syndrome (XFS) in Estonian patients scheduled for cataract surgery. METHODS: A series of 305 patients (mean age 71.5 +/- 8.6 years) scheduled for cataract surgery were examined for the presence of exfoliation syndrome, predominant type of lens opacification and intraocular pressure (IOP). RESULTS: Exfoliation material was detected in 108 (35.4%) patients, of whom 51 (47.2%) were bilaterally and 57 (52.8%) were unilaterally affected. In the bilaterally phakic patients exfoliation material was detected in 92 (30.2%) patients, of whom 51 (55.4%) were bilaterally and 41 (44.6%) were unilaterally affected. Intraocular pressure was higher in eyes with exfoliation than in eyes without it (19.2 +/- 6.5 mmHg and 17.1 +/- 3.8 mmHg, respectively; p = 0.006). Nuclear sclerosis predominated in eyes with XFS compared to those without XFS (57.6% and 36.9%, respectively). Cortical (7.6%) and subcapsular (7.6%) cataracts were less common in eyes with XFS than in eyes without XFS (16.9% and 20.2%, respectively). Mature cataract was found in 41.3% of all mixed types of cataracts. CONCLUSION: Exfoliation syndrome is common in Estonian patients. It was detected in 35.4% of the patients scheduled for cataract surgery. PMID- 15115446 TI - Consideration of the posterior corneal curvature for assessment of corneal power after myopic LASIK. AB - PURPOSE: To evaluate the effect of a separate measurement of the anterior and posterior corneal surface to calculate the total refractive power of the cornea after myopic laser in situ keratomileusis (LASIK). METHODS: A total of 39 eyes of 21 patients (aged 33 +/- 9 years) were included in this prospective, non randomized, comparative study. These involved 19 myopic corrections (- 3.5 +/- 1.6 dioptres) and 23 refractive corrections of myopic astigmatism (sphere: - 3.7 +/- 1.6 D, cylinder: - 1.2 +/- 0.4 D). All procedures were accomplished with the Keratom II). Coherent-Schwind excimer laser and the Moria Model One) microkeratome (150 micro m head) at the Medical Education Centre, La Trinidad, Caracas, Venezuela. Subjective refractometry, Bausch & Lomb) keratometry and Orbscan) slit-scanning corneal topography analysis were performed before and 3 months after LASIK. Corneal power was assessed directly using keratometry (K1) and Orbscan videokeratography (T1). Corneal power was calculated using the preoperative keratometric (K2, 'gold standard', clinical history method) or topographic power (T2, clinical history method) and spherical equivalent change. A composite value was derived from the Orbscan anterior and posterior surface power and central pachymetry (T3). RESULTS: Three months postoperatively, corneal power ranged in a descending order from T1 (42.33 +/- 1.78 D), K1 (40.82 +/- 2.20 D), K2 (40.42 +/- 2.36 D), T2 (40.03 +/- 2.51 D) to T3 (38.78 +/- 2.23 D). On average, T1 exceeded the gold standard by 1.9 D and the gold standard exceeded T3 by 1.6 D. K2, T1, T2 and T3 correlated significantly with K1 (r = 0.975, p < 0.001; r = 0.909, p < 0.001; r = 0.963, p < 0.001; r = 0.853, p < 0.001, respectively). The differences T1-K2 (r = - 0.699, p < 0.001) and T3-K2 (r = - 0.499, p = 0.001) correlated highly inversely and K1-K2 correlated borderline inversely (r = - 0.325, p = 0.043) with the intended refractive correction. CONCLUSION: After myopic LASIK, refractive corneal power is overestimated by direct keratometric and especially videokeratoscopic measurements. The higher the intended refractive correction, the greater is this error. A separate measurement of both refractive surfaces of the cornea tends to underestimate but may enhance accuracy of the total refractive corneal power if the history of the patient is unknown. PMID- 15115447 TI - Computerized calculation scheme for toric intraocular lenses. AB - BACKGROUND AND PURPOSE: While a number of intraocular lens (IOL) power prediction formulae are well established for determination of spherical lenses, no common strategy has been published for the computation of toric IOLs. The purpose of this study is to describe a paraxial computing scheme for tracing an axial pencil of rays through the 'optical system eye' containing astigmatic refractive surfaces with their axes at random. The capabilities of this computing scheme are demonstrated with clinical examples. METHODS: Based on a schematic model eye with spherocylindric surfaces, we use two alternative notations for description of vergences or prescriptions: (1) standard notation (refraction in both cardinal meridians and axis), and (2) component notation (spherical equivalent and cylindric component in 0 degrees and 45 degrees. Refractive surfaces are added to the vergence in component notation, whereas the transformation of the vergence through media is performed in the standard notation for both cardinal meridians. For calculation of the toric lens implant, a pencil of rays is traced through the spectacle and the cornea to the estimated lens position as well as backwards from the retina to the estimated lens position. For calculation of residual spectacle refraction, a pencil of rays is traced backwards from the retina through the toric lens implant and the cornea to the spectacle plane. RESULTS: In example 1 we calculate a 'thin toric lens' for compensation of a corneal astigmatism to achieve a spherical target refraction. In example 2 we compute a 'thick toric lens', which has to compensate for an oblique corneal astigmatism and rotate the spectacle cylinder to the against the rule position to enhance near vision. In example 3 we estimate the residual refraction at the corneal plane after implantation of a thick toric lens, when the cylinder of the lens implant is compensating the corneal cylinder in part and the axis of implantation is not fully aligned with the axis of the corneal astigmatism. CONCLUSION: This novel mathematical concept for computation of toric IOLs or prediction of the refractive outcome with a toric implant in place is a straightforward, computer based approach, which may substitute for more or less empirical methods of determining toric IOL implants. PMID- 15115448 TI - Amiodarone and optic neuropathy. AB - PURPOSE: To investigate the clinical, perimetric, contrast sensitivity and visual evoked potentials findings in patients on longterm treatment with amiodarone. METHODS: We enrolled 14 patients (age range 47-78 years, mean age 62.7 years) on chronic amiodarone therapy (mean cumulative dose of 519 g), and 14 age-matched control subjects. Participants were submitted to ophthalmological examination, kinetic visual field testing (Goldmann), standard achromatic automated perimetry (central 30-2, Humphrey), computerized isoluminant chromatic contrast sensitivity (CCS) and pattern visual evoked potentials evaluation (VEP). Data from baseline, 6-month and 1-year follow-up examinations were analysed. The main outcome measures were: initial and final corrected visual acuity; corneal, optic disc and fundus abnormalities; mean deviation (MD) and pattern standard deviation (PSD) central 30-2 perimetry parameters; visual field loss on kinetic perimetry; indices of CCS (tritan, protan and deutan axes), and pattern VEP latency (L-VEP) and VEP amplitude (a-VEP). RESULTS: We found a statistically significant difference between the patients and controls' baseline L-VEP and a-VEP measurements (p < 0.001). We found no significant correlation between prolonged L VEP and duration of amiodarone use (r = 0.07). After a 1-year follow-up, we evidenced an average increment in L-VEP of 1.96 ms and a-VEP measurements showed a decline of 0.69 micro V. In cumulative dose-specific analyses, mean differences between baseline and 12-month L-VEP measurements were more evident in those taking higher than 200 g cumulative amiodarone doses (p = 0.03). We found abnormal tritan CCS results in 19 eyes (68%). Of these, four eyes belonged to patients with no ocular disease (diabetes, cataracts). No significant differences were observed between the baseline and 12-month clinical eye examinations, central 30-2 perimetry and CCS results. CONCLUSION: We found a significant prolongation in pattern L-VEP and a significant reduction in pattern a-VEP in patients on longterm treatment with amiodarone compared to control subjects. Nevertheless, current data are insufficient to recommend appropriate methods of visual screening. PMID- 15115449 TI - Acupuncture treatment in patients with keratoconjunctivitis sicca: a pilot study. AB - AIM: To evaluate the effects of acupuncture in patients with keratoconjunctivitis sicca (KCS). MATERIAL AND METHODS: Twenty-five patients (20 women, five men) with KCS were randomly assigned to an acupuncture treatment group or a control group. The effects of acupuncture were evaluated by a questionnaire on symptoms, visual analogue scale recordings, registration of drop frequency, and dry eye tests. Ten acupuncture sessions were given. Follow-up was carried out after 2-3 weeks and again after a mean period of 8 months. RESULTS: Patients receiving acupuncture felt better at the first follow-up compared with the control group (p = 0.036). However, no statistical significance could be found concerning any change, or difference, in the total number of subjective symptoms, dosage frequency or, as indicated by the dry eye tests, tear quality, tear secretion and ocular surface disease. CONCLUSION: The results indicate that acupuncture has subjective beneficial effects in patients with KCS and could therefore be tried as a complement to ordinary treatment. PMID- 15115450 TI - Dry eye syndrome in thyroid-associated ophthalmopathy: lacrimal expression of TSH receptor suggests involvement of TSHR-specific autoantibodies. AB - PURPOSE: We evaluated the relationship between ocular surface damage, elevated lid aperture/impaired Bell's phenomenon and reduced tear production in thyroid associated ophthalmopathy (TAO). Suspecting a possible role of autoantibodies specific for TSH receptor (TSHR), we further investigated TSHR expression in the healthy lacrimal gland (LG). METHODS: A total of 48 patients with active TAO and 26 controls were examined for basal tear secretion, Rose Bengal and fluorescein staining, impression cytology (IPC), break-up time (BUT), and blinking (lid width, lid closure, ocular surface, upward excursion). Healthy LGs were investigated immunohistochemically for expression of TSHR. RESULTS: Thyroid associated ophthalmopathy patients showed significant ocular surface damage (Rose Bengal staining score: TAO: 2.0 [0-5] versus controls: 0 [0-0.4]; IPC score: TAO 3.0 [0-8] versus controls: 0 [0-1], and BUT: TAO: 3.0 seconds [0-9 seconds] versus controls: 19.5 seconds [13-35.4 seconds]), and significantly reduced tear secretion (TAO: 10 mm [3-20 mm] versus controls: 17 mm [10-27 mm]). Ocular surface damage correlated significantly with tear secretion (r = - 0.35) but not significantly with mechanical alterations (impaired upgaze [r = - 0.34] and ocular surface increase [r = 0.32]). We firstly demonstrate that lacrimal acinar cells physiologically express TSHR. CONCLUSIONS: As ocular surface damage in TAO significantly correlates with reduced tear production, LG impairment appears to be a major cause of ocular surface drying. Intriguingly, physiological expression of TSHR by LG suggests that, in thyroid disease, autoantibodies may bind to lacrimal TSHR and, perhaps via aberrant signal transduction, contribute to LG impairment and, hence, dry eye syndrome. PMID- 15115451 TI - Angiotensin-converting enzyme inhibitors (ACEIs) and age-related maculopathy (ARM): cross-sectional findings from the Blue Mountains Eye Study. AB - PURPOSE: To assess the relationship between the use of angiotensin-converting enzyme inhibitors (ACEIs) and prevalence of age-related maculopathy (ARM). METHODS: Eligible residents aged >/= 49 years were first examined in 1992-94 (Cross-section 1, n = 3654). Of these, 2335 were re-examined in 1997-99, together with an additional 1174 who became eligible after 1994 (Cross-section 2, n = 3509). Information regarding ACEI use was obtained and retinal photographs were graded using the Wisconsin ARM Grading System. RESULTS: In Cross-section 1, prevalence rates of late and early stage ARM were 1.3% and 4.3% among current ACEI users, and 2.0% and 4.8% among non-current users, respectively. In Cross section 2, prevalence rates of late and early stage ARM were 2.3% and 11.3% among current ACEI users, and 1.3% and 9.3% among non-current users, respectively. After adjusting for age, sex and smoking, neither survey found any significant association between ACEI use and prevalence of either late or early ARM. CONCLUSIONS: No significant cross-sectional associations were found between ACEI use and ARM prevalence in this population. PMID- 15115452 TI - Subacute visual loss from choroidal metastasis. PMID- 15115453 TI - Bilateral endogenous bacterial endophthalmitis: a report of four cases. AB - PURPOSE: To present and discuss four cases of bilateral endogenous endophthalmitis. METHODS: An observational study of four patients aged 55-80 years, seen within a 2-year period. All had diagnostic and therapeutic vitrectomy. The antibiotic therapy was guided by analyses of cultures of blood and vitreous. RESULTS: Blood cultures demonstrated Streptococcus pneumoniae in two patients and Staphylococcus aureus and Morganella morganii, each in one patient. The findings corresponded with culture findings from vitreous material in two patients. The primary foci for the metastatic spread of infection were endocarditis, discitis and a subdural abscess of the lumbar spine, urinary tract infection, and infection of a recent hip alloplasty, respectively, in the four patients. Five eyes became blind, whereas three eyes recovered to visual acuity of 0.25-0.67. CONCLUSIONS: Endogenous bacterial endophthalmitis usually leads to total loss of vision. The disease is acute and the time span for intervention limited. We believe that an active therapeutic approach including intravitreal antibiotics and vitreoretinal surgery saved three eyes from blindness. PMID- 15115454 TI - A pilot experiment using a network camera in ophthalmic teleconsultation. PMID- 15115455 TI - Bullous keratopathy treated with honey. PMID- 15115456 TI - Hypermetropic refractive change after hyperbaric oxygen therapy. PMID- 15115457 TI - Displacement of a laser in situ keratomileusis flap during retinal detachment surgery. PMID- 15115458 TI - Efficacy and safety of limbal anaesthesia for clear cornea phacoemulsification. PMID- 15115459 TI - Nd:YAG laser membranotomy for premacular haemorrhage. PMID- 15115460 TI - Recurrent retinal haemorrhages after glaucoma surgery. PMID- 15115461 TI - Uncommon presentation of asymmetrical retinopathy in diabetes type 1. PMID- 15115462 TI - Orbital involvement in multifocal fibrosclerosis. PMID- 15115463 TI - Myopic artists. PMID- 15115465 TI - Teenagers and cancer. PMID- 15115467 TI - A literature review of head and neck cancer patients information needs, experiences and views regarding decision-making. AB - This study reviews the body of existing research undertaken to explore head and neck cancer patients experiences of treatment for surgery, their information needs and their views regarding decision-making. Patients who suffer head and neck cancers and undergo surgery often report considerable psychological distress and impaired social functioning. To optimize survival, the decision about what treatment option to follow is often made quickly, with little support in terms of counsel or the provision of information. It is suggested that there is inadequate previous work exploring the content and delivery of information required by patients, their experiences and their views regarding decision-making to guide appropriate clinical interventions. By assimilating and critiquing existing work in this review an appropriate focus for further research is likely to be determined. An electronic search of relevant databases was undertaken including Cinahl, Medline, Psychinfo, Assia, the Cochrane Library and British Nursing Index. In addition, a grey literature search was also undertaken by hand. The search strategy was developed using recommendations from the Centre for Reviews and Dissemination 'Guidance for Those Carrying Out or Commissioning Reviews' (CRD Report Number 4, 2001). There is significant psychosocial and other functional disturbance amongst survivors of head and neck cancer surgery. Improved information giving may play a part in determining these outcomes. PMID- 15115468 TI - The concepts of fatigue and depression in cancer. AB - A strong association between fatigue and depression in cancer patients has been reported repeatedly in clinical studies. The distinction remains difficult, mainly because of the similar phenomenology of fatigue and depression. It is the aim of this paper to work out similarities and differences in the conception of fatigue and depressive disorders. For that, a differentiation between depression as emotional distress and depression as clinical syndrome, according to the current classification systems, has to be made. Therefore, the classification of depressive disorders and their criteria is presented in the second section of this paper, especially in view of the diagnosis of depressive disorders in cancer patients. The comparison of the multidimensional fatigue construct and depression shows a strong overlap of symptoms. None of the fatigue symptoms are specific for fatigue, all being elements of depressive syndromes. It is in particular the psychological symptoms of depressive disorders that differentiate between the two concepts. To that end, the question is discussed whether fatigue in its current conceptualization can be defined as a diagnostic entity independent of depressive disorders. Additionally, research approaches are presented from the area of the chronic fatigue syndrome and neurasthenia, which could be adapted to cancer related fatigue and help to clarify the clinical differences between fatigue and depression. In order to ensure better differential diagnostics in the future, criteria-orientated research in particular is needed. PMID- 15115469 TI - The European Organization for Research and Treatment of Cancer QLQ-C30: an examination into the cultural validity and reliability of the Turkish version of the EORTC QLQ-C30. AB - The Turkish version of the European Organization for Research and Treatment of Cancer Quality of Life Core Questionnaire version 2.0 (EORTC QLQ-C30 v.2.0) has started to be used in clinical trials recently. The objective of the study was to evaluate the validity and reliability of the Turkish version of the EORTC QLQ-C30 v.2.0 and the correlation between the Karnofsky Performance Scale (KPS) and the EORTC QLQ-C30. Two hundred and two lung cancer patients were included in the study between January and March 2000. All the subscales met the minimal standards of reliability (Cronbach's alpha > or = 0.70). Only the role functioning scale differed among the three disease stages of patients (local, locoregional and metastatic). There was no statistically significant difference among therapy types. All interscale correlations were statistically significant (P < 0.01). The strongest correlations were found among the physical functioning, role functioning and fatigue scales. Social functioning was closely related with physical, role, emotional and cognitive functioning. The weakest correlations were between nausea/vomiting and the other scales. Global quality of life (QOL) was substantially correlated with most of the scales except cognitive functioning. The coefficients for the correlation between the items differed between 0.12 and 0.97 and all the subscales were strongly correlated with the scales which they formed. The highest correlation between the EORTC QLQ-C30 and KPS was for physical functioning (r = 0.62, P < 0.05). The Turkish version of the EORTC QLQ-C30 is a valid (by means of interscales validity) and reliable instrument for Turkish lung cancer patients and can be used in clinical studies but needs supporting by the reference data on the QOL of the Turkish population. PMID- 15115470 TI - What is it like living with the diagnosis of cancer? AB - This article discusses in-depth interviews of 12 persons, who had been diagnosed for cancer from 6 months to 3 years earlier, concerning what it is like to live with the diagnosis of cancer. The persons interviewed were aged 43-70 years, and 11 of the 12 had received successful radical treatment; one patient was treated for symptoms, but with an uncertain future development. This investigation shows that the period of waiting from the first suspicion of cancer until diagnosis confirmation of the disease was a period of great stress, and that the following period until the commencement of treatment was also a very difficult one. The investigation also shows that there was no follow-up of individual patients concerning their need to talk about their difficult new life-situation brought about by the disease. Receiving the diagnosis of cancer was remembered as being a very dramatic experience for the informants. The informants were satisfied with the medical treatment they received, but they also feel that there should have been some kind of programme available to care for their psycho-social needs. In the rehabilitation phase, they feel that they were left to their own resources; during this period, they felt physically and mentally drained, and they did not have the knowledge to tackle their new and difficult life-situation. The threat of new cancer cells being discovered is always with patients. Attending the check ups is also described as a period of stress. PMID- 15115471 TI - Cancer and communication: similarities and differences of men with cancer from six different ethnic groups. AB - This paper reports the communication aspects of a pilot study, which explored the cancer meanings and experiences of six men with cancer and their significant others from different ethnic groups. A case study design was applied using the principles of phenomenology. In-depth semi-structured individual interviews were conducted in participants' own homes, in London, UK. This paper will only deal with the communication aspects of the findings. Ten themes emerged from the comparative analysis of the study's data, with communication as a cross-cutting theme. Further analysis of this theme revealed similarities and differences of the participants' experiences of communicating with health professionals; families or friends; and God/Allah. In addition, similarities and differences in communicating meanings of cancer in different cultures were revealed. The findings revealed similarities in the way that men from these six cultures communicate with health professionals and their families following a diagnosis of cancer, and differences in how they communicated with God/Allah, which depended on their religious beliefs and practices. PMID- 15115472 TI - Patient satisfaction: a descriptive study of a breast care clinic in Iran. AB - A descriptive study was carried out to examine patient satisfaction among women attending the Iranian Centre for Breast Cancer. A specially designed patient satisfaction questionnaire was distributed to all attendees and they were asked to complete the questionnaire. The questionnaire contained items on satisfaction with care organization, physical environment, personnel communication skills, clinical care, and overall satisfaction. In all, 425 women participated in the study. The mean age of women was 40.4 years (SD = 11.6), most were married (81%) and housewives (69%). A vast majority of women were very satisfied or satisfied with physical environment, personnel communication skills, and clinical care received. Eighty-two per cent of respondents were very satisfied or satisfied with the clinic's overall performance. There was a significant agreement between patients' overall satisfaction and satisfaction with physical environment, personnel communication skills, and clinical care. There was greatest agreement between patients' overall satisfaction and satisfaction with examination room (Kappa = 0.21, P < 0.0001) and with physicians' consultation (Kappa = 0.20, P < 0.0001). None of the demographic variables showed any significant association with patients' overall satisfaction. The findings suggest that the physical environment and physicians' style of consultation contribute most to the patients' overall satisfaction. PMID- 15115473 TI - Healthy women from suspected hereditary breast and ovarian cancer families: the significant others in their lives. AB - This paper investigates communication and interactions between healthy women from families with a history of breast/ovarian cancer and five statuses of significant others: (1) women friends; (2) sisters; (3) brothers; (4) male partners; and (5) children in order to better understand the way the family deals with cancer genetics risk information and the extent of social support available to its members. We conducted a research ethics committee reviewed exploratory, qualitative study at a major clinical and research cancer centre in the United Kingdom from January to June 2000. Twenty-one semi-structured, in-depth interviews were conducted using a purposive sample of women coming to the cancer genetics risk clinic for the first time, supplemented by 5 months of participant observation. On the whole, women friends consistently provided strong social support. Sisters were usually close, but communication about the breast/ovarian cancer in their family in some cases was quite limited and fraught with emotional overtones. Brothers were the most difficult to relate to regarding cancer in the family and seemed almost to exist in a different 'interrelational space'. The women claimed that their male partners were supportive, but with caveats. Mothers worried about how much information and at what age they should inform their children about the specifics of the family history of breast/ovarian cancer and tried to protect them when they were young. The women were very concerned about their daughters and granddaughters, but were far less concerned about the impact on their sons. PMID- 15115474 TI - Loss to follow-up of patients with malignant lymphoma. AB - Loss to follow-up (LTFU) in cancer patients is a serious problem, yet there is little data on this and on the underlying reasons. Of 144 paediatric and 431 adult patients with lymphoma diagnosed in 1997/1998 at King Faisal Specialist Hospital and Research Center, Riyadh (KFSHRC), 30% and 48.5%, respectively, were LTFU after 4 years (excluding patients known to have died). In 2001-2002, 196 paediatric and adult lymphoma patients at KFSHRC were enrolled in a prospective study in which explanations were obtained in detail for non-attendance at follow up appointments (No Show). Sixteen months after commencement of the study, 49 patients were No Show, because of patient-based communication problems (20), transportation problems (8), patient not contactable (18), and personal reasons (3). In addition, patients were recorded incorrectly as No Show through hospital/patient communication problems. The No Show patients, especially the 23 who failed to keep a second appointment, are identifiable as potential LTFU during the 3 years in which this cohort will be followed. This study and, we suggest, other studies on LTFU should stimulate interest in this issue, in the predisposing factors, and in strategies to address them. PMID- 15115475 TI - Provision of oncology services in remote rural areas: a Scottish perspective. AB - There is a paucity of research into rural health care services. In particular little is known about the provision of specialist cancer services for patients who live in remote rural areas of the UK. This study set out to investigate current models of medical and clinical oncology care in Scotland. A national survey with key health professionals was conducted to identify rural oncology schemes currently in operation. Detailed quantitative data about the schemes together with qualitative data on how health professionals view current models of care were collected by a computer-assisted telephone survey. Schemes that currently provide outpatient and chemotherapy oncology services for remote rural patients fell into three categories: central clinics (5); shared care outreach clinics with chemotherapy provision (11); and shared care outreach clinics without chemotherapy provision (7). All radiotherapy was conducted at central clinics (5). Widely varying practices in delivery of cancer care were found across the country. The main issues for professionals about current models of care involved expertise, travelling and accessibility (for patients), communication and expansion of the rural service. Nation-wide consistency in cancer care has still to be achieved. Travelling for treatment was seen to take its toll on all patients but particularly for the very remote, elderly and poor. Most professionals believe that an expansion of rural services would be of benefit to these patients. It is clear, however, that the proper infrastructure needs to be in place in terms of local expertise, ensured quality of care, and good communication links with cancer centres before this could happen. PMID- 15115476 TI - Study of white coat effect on the prognosis of women with breast cancer. PMID- 15115480 TI - Abstracts of the IX World Congress of the International Society for Diseases of the Esophagus. PMID- 15115497 TI - The urticaria spectrum: recognition of clinical patterns can help management. AB - Urticaria has diverse clinical presentations and causes. The implication of classifying urticaria primarily by clinical presentation rather than aetiology is that management can be focused on specific clinical problems without extensive investigations. Management pathways may involve nonpharmacological measures and drug interventions, which can be grouped into first-, second- and third-line therapies. Stronger, but potentially more risky, second- and third-line approaches may be justified for patients who do not respond to first-line therapy with antihistamines even though it may not be possible to define a specific aetiology, such as autoimmune urticaria, with confidence. PMID- 15115498 TI - Vascular anomalies in Proteus syndrome. AB - Proteus syndrome (PS) is a complex hamartomatous disorder defined by local overgrowth (macrodactyly or hemihypertrophy), subcutaneous tumours and various bone, cutaneous and/or vascular anomalies (VA). VA are manifold in PS, but their prevalence is unknown so far. In order to further characterize PS, we studied the prevalence of VA in 22 PS patients presenting to our outpatient clinic and reviewed 100 PS patients previously reported between 1983 and 2001. The diagnosis of vascular abnormalities was made on clinical grounds and supported with imaging studies and/or histology in 12 and seven patients out of 22, respectively. Thirty five VA were identified in 22/22 (100%) of our patients, and more than one type of VA were present in 10 of them. Vascular tumours, portwine stains (PWS), and venous anomalies (varicosities, prominent veins) were equally common. A total of 118 VA were previously reported in 70/100 (70%) PS patients; vascular hamartomas were more prevalent (56/118 = 47.5%), whilst PWS (21.2%) and venous anomalies (22.9%) were slightly less common than in our series, but there is the possibility of under-reporting. Unlike Klippel-Trenaunay syndrome, where VA are mostly confined to the hypertrophic limb, major arteriovenous anomalies are rare, and - similar to the other hamartomas and naevi observed in PS (pigmentary naevi, epidermal naevi, subcutaneous tumours, exostoses) - VA appear to be distributed at random sites on the body. We conclude that VA are among the most common findings in PS. Their varying type and distribution lend further support to the concept of somatic mosaicism. PMID- 15115499 TI - A randomized cross-over study to compare PUVA and extracorporeal photopheresis in the treatment of plaque stage (T2) mycosis fungoides. AB - PUVA is a well-established and effective treatment for plaque stage mycosis fungoides (MF) but its use is limited on a long-term basis because of the risk of cutaneous carcinogenesis. A further disadvantage is that nonexposed areas (sanctuary sites) often develop persistent disease. Therefore it is important to find alternative methods of treatment. Extracorporeal photopheresis (ECP) is a form of photochemotherapy that involves exposure of white blood cells to UVA with psoralens and can be effective in Sezary syndrome and erythrodermic cutaneous T cell lymphoma. The aim of this study was to compare the efficacy of PUVA and ECP in the treatment of patients with T2 plaque stage (Stage 1B) MF who had a detectable peripheral blood T-cell clone. The study was of a cross-over design. Sixteen patients were randomized to receive either PUVA twice weekly for 3 months followed by ECP once monthly for 6 months at relapse, or vice-versa. Response was assessed by monthly skin scores and peripheral blood T-cell clonality. Ten patients received PUVA initially and six ECP initially. Eight patients completed the study. Skin scores taken at the completion of each treatment arm in patients who completed the study were 113 units better (confidence interval, 42-184 units) following 3 months PUVA than 6 months ECP (P = 0.002). Peripheral blood T-cell clones were detectable in all patients post-treatment. This study indicates that ECP is not effective in the treatment of plaque stage (1B/T2) MF even in patients with molecular evidence of a peripheral blood T-cell clone. Although PUVA was more effective than ECP, neither treatment modality cleared malignant T-cells from the peripheral blood. PMID- 15115500 TI - Efficacy of imiquimod 5% cream for basal cell carcinoma in transplant patients. AB - Imiquimod 5% cream has proven to be effective in superficial and nodular basal cell carcinomas in nonimmunosuppressed patients and treating squamous cell carcinomas in situ in transplant patients. The objective of this open-label study was to determine the efficacy of imiquimod 5% cream in treating basal cell carcinoma in transplant patients. At our unit, four renal transplant patients and one cardiac transplant patient were diagnosed with 10 basal cell carcinomas in 2001. Four tumours were superficial, three nodular and three infiltrative. Five basal cell carcinomas received imiquimod 5% cream at night four times weekly for 6 weeks, without occlusion, and the other five tumours were treated on 5 nights per week for 5 weeks. Biopsies taken 6 weeks after the end of treatment showed no tumour in seven of 10 of the cases. Notably, all four superficial basal cell carcinomas, two of the three of nodular lesions and one of the three of infiltrative cases had completely cleared. PMID- 15115501 TI - Kikuchi's disease (necrotizing lymphadenitis) with cutaneous involvement associated with subacute cutaneous lupus erythematosus. AB - Necrotizing histiocytic lymphadenopathy (Kikuchi's disease) is a rarely observed clinical entity characterized by fever, and solitary or multiple lymphadenopathy predominantly in the posterior cervical region. Kikuchi's disease has been reported to precede, coexist with or follow the diagnosis of systemic lupus erythematosus. In only rare instances has its association with cutaneous lupus erythematosus without systemic involvement been reported. We report a 45-year-old woman who presented characteristic systemic and cutaneous manifestations of Kikuchi's disease. Several months later, after sun exposure, she developed lesions of subacute cutaneous lupus erythematosus. The American Rheumatism Association criteria for systemic lupus erythematosus were not fulfilled. The possible pathogenic relationships between the two processes are discussed. PMID- 15115502 TI - Extracorporeal photochemotherapy for the treatment of exanthematic pityriasis rubra pilaris. AB - Pityriasis rubra pilaris (PRP) is a rare papulosquamous skin disease of unknown aetiology that has been categorized into five clinical types based on age at onset, cutaneous features and prognosis. We present a patient with chronic exanthematic type II atypical adult PRP, whose skin status was significantly improved with monthly extracorporeal photochemotherapy (ECP). Various therapeutic regimens including narrow-band UV-B, bath PUVA therapy, systemic fumaric acid esters and systemic cyclosporin had failed. Oral retinoids could not be administered due to a type IIa hyperlipoproteinemia with profound hepatic steatosis and elevated liver transaminases. The observed clinical benefit may encourage future clinical studies analysing the effectiveness of ECP in otherwise unresponsive cases of type II PRP. PMID- 15115503 TI - Association of verrucous skin lesions and skin ulcers on the feet in patients with diabetic neuropathy. AB - We report the simultaneous or chronological association of verrucous skin lesions and diabetic ulcers on the feet of three diabetic patients. All three patients had poor diabetic control and were suffering from complications such as neuropathy, retinopathy and nephropathy at the time of presentation. In patient 1, verrucous skin lesions on the feet in diabetic neuropathy (VSLDN) and a diabetic skin ulcer developed simultaneously. In patient 2, VSLDN preceded the development of diabetic ulcers, while in patient 3, diabetic ulcers preceded VSLDN. These associations suggest that VSLDN and diabetic ulcers are closely related in their aetiology and pathogenesis. Strategies for the treatment and prevention of VSLDN should include multiple treatment modalities combined with foot care as proposed by the international working group on the diabetic foot. PMID- 15115504 TI - Disseminated cutaneous infection with Mycobacterium chelonae in a patient with steroid-dependent rheumatoid arthritis. AB - Mycobacterium chelonae is a rapidly growing atypical mycobacterium that is a normal commensal of water and soil. We report a case of a 61-year-old man with seronegative rheumatoid arthritis and fibrosing alveolitis on long-term prednisolone who presented with a number of tender, red, subcutaneous nodules on his upper arms and a pustule on his left cheek. Histopathologic examination revealed dense neutrophilic collections within the deep dermis and subcutaneous fat with abscess formation. Long filamentous organisms were seen within these collections and were subsequently identified by special stains and PCR as Mycobacterium chelonae. Treatment was not possible as the patient developed bacteria bronchopneumonia before identification of the organism and he subsequently died. Post-mortem revealed no extra-cutaneous evidence of mycobacterium infection. PMID- 15115505 TI - Nephrogenic fibrosing dermopathy. AB - Nephrogenic fibrosing dermopathy (NFD) is an uncommon, recently described fibrotic skin disorder that has been observed in patients with end-stage renal disease. We describe a 81-year-old man with end-stage renal disease who developed NFD in the seventh year of haemodialysis. Laboratory investigations revealed considerably increased circulating immune complexes and anti double stranded DNA antibodies. Because the pathogenesis of NFD may not entirely be attributable to haemodialysis, the role of circulating factors in the development of cutaneous manifestations requires further investigation. PMID- 15115506 TI - Intestinal bypass syndrome presenting as erythema nodosum. AB - Intestinal bypass was a popular surgical procedure for morbid obesity resulting, on average, in a 50 kg weight loss. We describe a 66-year-old woman who underwent the procedure 12 years earlier and subsequently presented with recurrent episodes of erythema nodosum-like lesions. Further investigations revealed hyperoxaluria, renal failure, deficiency of fat-soluble vitamins (causing night blindness, osteomalacia and easy bruising) and anaemia. Antibiotics led to only temporary remission and, as with 24-30% of similar cases, she underwent surgical reversal to prevent the complications from worsening. PMID- 15115507 TI - Bullous subacute cutaneous lupus erythematosus. AB - We describe a 59-year-old woman, with a history of autoimmune disease and disseminated uterine leiomyosarcoma, who developed a photoaggravated, blistering skin eruption. An initial rash, at the outset of treatment with chemo- and radiotherapy, resembled erythema multiforme. Review of the original skin biopsy showed it to be subacute cutaneous lupus erythematosus. There were no systemic symptoms or signs to suggest systemic lupus erythematosus. The much later photoaggravated rash consisted mainly of bullae and eventual epidermal denuding which resembled toxic epidermal necrolysis. We propose that the clinical and histological diagnosis is one of bullous subacute cutaneous lupus erythematosus in a patient with no other features of systemic lupus erythematosus. PMID- 15115508 TI - Recalcitrant erosive flexural lichen planus: successful treatment with a combination of thalidomide and 0.1% tacrolimus ointment. AB - A 79-year-old woman has had chronically eroded and ulcerated flexural lichen planus for 12 years, resistant to many forms of treatment. She was successfully treated initially with a combination of topical 0.1% tacrolimus ointment and oral thalidomide and then with topical tacrolimus alone. She has remained free of exacerbations for 12 months and treatment has been well tolerated. Erosive lichen planus involving flexures alone is rare. All reports on treatment of this condition address erosive oral or mucosal lichen planus and both thalidomide and topical tacrolimus have been reported individually to be beneficial. PMID- 15115509 TI - Beneficial effects of topical tacrolimus on recalcitrant erosions of pemphigus vulgaris. AB - We report a case of pemphigus vulgaris in which a recalcitrant area of erosion on the cheek cleared only when topical tacrolimus was used in addition to a regime of systemic therapy consisting of cyclophosphamide and prednisolone. Clinical improvement occurred within 10 days of applying topical tacrolimus with healing of erosions and reduction in pain and burning sensations. Topical tacrolimus may inhibit local activation of T lymphocytes through altered expression of cytokines such as interleukin-1, -4 and -5, tumour necrosis factor-alpha and interferon gamma. Some of these cytokines may also contribute directly to increasing keratinocyte fragility in the aetiology of pemphigus vulgaris erosions. This case illustrates that topical tacrolimus may be a useful adjunct in the management of patients with pemphigus vulgaris. PMID- 15115510 TI - Pruritus in adult dermatomyositis. AB - Dermatomyositis has a significant clinical component of pruritus that has not yet been studied. Pruritus can significantly affect the life of patients. The aim of the present work was to study the degree of pruritus experienced by patients. A four-question survey was sent to patients with documented dermatomyositis. The survey used a 100-mm Visual Analogue Scale (VAS) to describe current, worst and daily pruritus, and the effect this has on daily activities. Twenty-six subjects returned completed questionnaires: four had no pruritus; the majority had a significant amount with means above 50 on the VAS. A mean of 44.6 was found for the effect on daily life. Further studies should be performed to examine the prevalence and severity of pruritus in this population and it's effect on their quality of life. Clinicians must be aware of the significant pruritus and provide adequate therapy to improve quality of life. PMID- 15115511 TI - Urinary leukotriene E4 correlates with severity of atopic dermatitis in children. AB - Leukotriene E4 (LTE(4)) is elevated in adults with atopic dermatitis (AD). We evaluated whether urinary LTE(4) as a noninvasive marker correlates with clinical indices of disease activity in children with AD. AD patients aged 18 years or younger were eligible for inclusion in the study. Disease severity over the preceding 3 days was evaluated by the SCORing Atopic Dermatitis (SCORAD) index. Severity of AD over the past 12 months was evaluated by the Nottingham Eczema Severity Score (NESS) in Chinese. Urinary LTE(4) concentration was measured by competitive enzyme immunoassay. One hundred and twenty-six children with AD (82 boys and 44 girls) and 45 controls were recruited. The mean +/- SD urinary log transformed LTE(4) concentration in AD patients and controls was 2.94 +/- 0.32 and 2.62 +/- 0.20 pg/mg creatinine, respectively (P < 0.0001). SCORAD significantly correlated with NESS (r = 0.681, P < 0.0001). There were significant correlations between urinary LTE(4) concentration and overall SCORAD score (r = 0.270, P = 0.002) and its extent (r = 0.185, P = 0.038) and intensity components (r = 0.247, P = 0.005), but not with NESS. When compared with mild AD, urinary LTE(4) concentrations were higher in patients with moderate-to-severe disease (P = 0.049). Urinary LTE(4) measurement is noninvasive and may be useful in supplementing the SCORAD for following longitudinal changes in AD severity in children. However, the practical value of this assay in a clinical setting remains to be determined. PMID- 15115512 TI - Anti-inflammatory effect of topical nanocrystalline silver cream on allergic contact dermatitis in a guinea pig model. AB - The anti-inflammatory activity of topical nanocrystalline silver cream was assessed and compared with the effects of topical steroids and currently available immunosuppressants using a guinea pig model of allergic contact dermatitis. Dermatitis was induced with dinitrochlorobenzene and treated with different concentrations of nanocrystalline silver, medium and high potency steroids, tacrolimus and pimecrolimus, or appropriate vehicles once daily for 5 days. Erythema was evaluated daily (on a score of 0 to 4, from absent to very severe) and histopathology of the skin biopsies was evaluated after 5 days of treatment. Prior to treatment, the average scores of erythema in all the groups were in the range of 3(+) to 4(+). In the no treatment and vehicles groups these scores remained at about this level for the subsequent 5 days of the study. Nanocrystalline silver reduced erythema within 1 day of treatment in a concentration-dependent manner with significant reduction at silver concentrations of 0.5% and 1% (P < 0.05) and this reduction progressed throughout the study period. Steroids and immunosuppressants produced similar decreases in erythema, with no significant differences compared to 0.5% and 1% nanocrystalline silver. In skin biopsies scored for degree of inflammatory response, effects of treatments mirrored erythema results. This study suggests that nanocrystalline silver cream may have therapeutic potential for topical treatment of inflammatory skin diseases. PMID- 15115513 TI - The distribution of IgG subclasses in the lupus band suggests disease-specific alteration in subclass switching rather than polyclonal B-cell activation. AB - Deposition of immunoglobulins in the skin of patients with lupus erythematosus (LE), demonstrable as a linear band 'lupus band' at the basement membrane zone (BMZ) by direct immunofluorescence, was first described in 1963. Four decades after the discovery of the lupus band, a basic question regarding the origin of immunoglobulins of the lupus band is still unanswered. Is the lupus band just a manifestation of polyclonal B-cell activation commonly seen in systemic LE (SLE)? The distribution of IgG subclasses deposited in the skin of patients with SLE was identified using immunohistochemistry. The relative restriction of IgG of the lupus band to the IgG3 subclass demonstrated in this study provides evidence against polyclonal B-cell activation as the only cause of the lupus band and suggests disease-specific alteration in subclass switching. PMID- 15115514 TI - Serum levels of cutaneous T-cell attracting chemokine (CTACK) as a laboratory marker of the severity of atopic dermatitis in children. AB - There are at least 13 scoring systems for the assessment of disease severity in atopic dermatitis (AD). Each system has its problems with interobserver and intraobserver variability. Cutaneous T-cell attracting chemokine (CTACK) is a skin-specific chemoattractant which may correlate with AD severity and obviate the issue of observer reliability. We evaluated whether serum CTACK concentrations were associated with the severity of AD in children according to the SCORing Atopic Dermatitis (SCORAD) index. Thirty-seven Chinese children with AD (23 boys, 14 girls; aged 1-11 years) and 13 controls were recruited. The median (interquartile range) overall SCORAD for AD patients was 29.7 (20.3-49.7). Serum concentrations of CTACK and two other atopy-related chemokines, macrophage derived chemokine (MDC) and thymus and activation-regulated chemokine (TARC), were measured by sandwich enzyme immunoassay. There were significant correlations between SCORAD (r = 0.394, P = 0.016), its area (r = 0.528, P = 0.001) and intensity components (r = 0.429, P = 0.008) with serum levels of CTACK. The serum concentrations of inflammatory markers MDC and TARC also correlated with the CTACK concentrations (r = 0.618, P < 0.001, and r = 0.587, P = 0.001, respectively). Serum CTACK concentration appears to be a skin-specific objective marker that correlates with various clinical and laboratory parameters of AD. PMID- 15115515 TI - Remission of rosacea induced by reduction of gut transit time. AB - Rosacea is a chronic disorder characterized by hypersensitivity of the facial vasculature, presenting with intense flushing eventually leading to chronic erythema and telangiectasia. Although the precise aetiology of rosacea is not known, numerous associations with inflammatory gastrointestinal tract disorders have been reported. Furthermore, substance P-immunoreactive neurones occur in considerably greater numbers in tissue surrounding affected blood vessels suggesting involvement of neurogenic inflammation and moreover plasma kallikrein kinin activation is consistently found in patients. In this report, a patient without digestive tract disease is described, who experienced complete remission of rosacea symptoms following ingestion of a material intended to sweep through the digestive tract and reduce transit time below 30 h. It is possible that intestinal bacteria are capable of plasma kallikrein-kinin activation and that flushing symptoms and the development of other characteristic features of rosacea result from frequent episodes of neurogenic inflammation caused by bradykinin induced hypersensitization of facial afferent neurones. The possible relevance of this hypothesis to other conditions featuring afferent hypersensitivity, such as fibromyalgia, is considered. PMID- 15115516 TI - Treatment of Microsporum spp. tinea capitis with pulsed oral terbinafine. AB - Oral terbinafine is widely used in the treatment of superficial dermatomycoses as well as subcutaneous and systemic mycoses. It is also useful in treating tinea capitis, although for Microsporum canis and some ectothrix organisms, the effectiveness of the drug may be less than for some endothrix infections. In this study, we undertook a double-blind randomized trial comparing standard and double doses of terbinafine given in a pulsed protocol (1 week on, 3 weeks off) in treating Microsporum spp. tinea capitis in 42 individuals. We found that pulsed terbinafine at a higher dose did not improve treatment efficacy. However, our data clearly demonstrated that the duration of treatment is an important factor in determining clinical outcome and cure. Two pulses of standard dose terbinafine were found to be sufficient for treating most cases of Microsporum spp. tinea capitis, although additional treatment (a third pulse) may be needed if clinical improvement is not evident at 8 weeks after initiating therapy. PMID- 15115517 TI - A novel missense mutation in the COL7A1 gene underlies epidermolysis bullosa pruriginosa. AB - Epidermolysis bullosa (EB) pruriginosa is a subtype of dominant dystrophic EB (DDEB), characterized by severe pruritus and blistering localized to the extensor surface of the extremities. EB pruriginosa exhibits extensive clinical heterogeneity with variable expression and delayed age of onset. Mutations in the COL7A1 gene, especially in glycine residues within Gly-X-Y repeats, have been shown to cause this form of DDEB. Here, we report a novel COL7A1 mutation in a Taiwanese pedigree with EB pruriginosa. Using PCR and direct sequence analysis we have identified a G-->T transversion at nucleotide 7097 in exon 92 of COL7A1, converting a glycine residue to valine (G2366V). The mutation resides within a consecutive, uninterrupted stretch of 17 Gly-X-Y residues in the triple-helical domain of type VII collagen. Interestingly, an affected member of this family also displayed elevated IgE levels, previously reported in some patients with this disorder. Our finding further implicates COL7A1 mutation in the pathogenesis of EB pruriginosa and underscores the heterogeneous clinical symptoms of glycine mutations in DDEB. PMID- 15115518 TI - A novel keratin 9 gene mutation (Asn160His) in a Taiwanese family with epidermolytic palmoplantar keratoderma. AB - Epidermolytic palmoplantar keratoderma (EPPK) is an autosomal dominant inherited disorder of keratinization. Recent molecular studies have shown that EPPK is caused by mutations in keratin 9 gene (KRT9). We report a Taiwanese family with EPPK with a novel mutation with an A-->C transition at the first nucleotide of codon 160 in KRT9. The mutation is predicted to result in an asparagine to histidine substitution (N160H) at the beginning of the alpha-helical 1A domain of keratin 9. Mutations in this region could disrupt keratin filament assembly, leading to degeneration or cytolysis of keratinocytes. Our mutation analysis confirms that codon 160 in KRT9 is one of the mutation hot spots in EPPK. PMID- 15115519 TI - Epidermodysplasia verruciformis-like eruption associated with HIV infection. PMID- 15115520 TI - Laugier-Hunziker Syndrome. PMID- 15115521 TI - Two Singaporean cases of guttate leucoderma in Darier's disease. PMID- 15115522 TI - Crusted scabies as a cause of longitudinal nail splitting. PMID- 15115523 TI - Autoimmune progesterone dermatitis presenting with purpura and petechiae. PMID- 15115524 TI - Successful treatment of scleromyxoedema with high dose intravenous immunoglobulin. PMID- 15115525 TI - Demodicidosis in scalp rosacea? PMID- 15115526 TI - Subacute cutaneous lupus erythematosus associated with leflunomide. PMID- 15115527 TI - Asymmetric periflexural exanthem in an adult. PMID- 15115528 TI - Diffuse cutaneous systemic sclerosis treated with intravenous iloprost. PMID- 15115529 TI - Which term should be used to describe drug eruptions confined to sites of previous herpes zoster lesions, 'isotopic response' or 'recall phenomenon'? PMID- 15115531 TI - Is hypoallergenic a credible term? AB - Hypoallergenic is a term that is frequently applied to cosmetics and is intended to imply a very low likelihood of developing or potentiating allergic cutaneous reactions. However there are neither industry standards nor legal requirements which must be fulfilled in order to make this claim. We describe a 25-year-old woman referred with worsening eczema which she related to the use of two skincare products. Both had been promoted as hypoallergenic and 'preservative free'; chemical analysis using HPLC confirmed the presence of methyldibromoglutaronitrile and formaldehyde, both at robust concentrations. We consider it unlikely that these were present as contaminants and suggest that the term hypoallergenic must be interpreted with caution. PMID- 15115532 TI - Variations in registration of skin cancer in the United Kingdom. AB - Cancer registries are used to compare incidences between regions, plan for service provision, and to assess the impact of health interventions. Significant variation in data capture for skin cancers and reporting of results was evident between regional cancer registries in the UK when assessed in 1991. Using a postal questionnaire we sought to document methods of recording skin cancer incidence in the UK in 2000, and to assess if practice has changed from 1991. All UK cancer registries were asked for details of their method of skin cancer case registration and latest available incidence figures. Methodology was assessed against recently implemented national standards. All registries responded to the survey. Sources of data were more uniform than was the case 9 years ago. All registries except one attained national standards for basal cell carcinoma data collection, but only half of the registries attained standards for squamous cell carcinoma. Ten of the 12 correctly recorded numbers of malignant melanomas, but three still failed to record the Breslow thickness or Clarke's level. Wide variation is evident in the recorded incidences for each of the malignancies, and the efficiency at which figures are made available. Thus, although there has been improvement since 1991, variability still exists between UK registries in methods of data capture, the data recorded, and efficiency of data processing in skin cancer registration. PMID- 15115533 TI - Case 1. Diffuse melanonychia. PMID- 15115534 TI - Case 2. Porokeratotic eccrine and ostial dermal duct naevus. PMID- 15115535 TI - Case 3. Brooke-Spiegler syndrome. PMID- 15115536 TI - Learning dermatology. AB - Students should be involved in curriculum planning and encouraged to formulate their own learning outcomes. Adult learners appreciate opportunities to learn from real or simulated experiences that are applicable to real-life situations. Contact with patients is essential to help students to develop empathy and communication skills and patients with chronic skin conditions such as psoriasis can make an active contribution to the education of students. Students learning dermatology use both pattern- recognition and analytical processes, but pattern recognition develops only with repetition and it is not realistic to expect students to acquire many diagnostic skills. Instead teachers should provide students with a framework that will enable them to analyse skin conditions, to understand the relevance of what is taught and to apply their learning in the future. All medical graduates should be able to describe the skin and record their findings each time they clerk a patient. PMID- 15115537 TI - A review of recent journal highlights. PMID- 15115540 TI - Identification of "pathologs" (disease-related genes) from the RIKEN mouse cDNA dataset using human curation plus FACTS, a new biological information extraction system. AB - BACKGROUND: A major goal in the post-genomic era is to identify and characterise disease susceptibility genes and to apply this knowledge to disease prevention and treatment. Rodents and humans have remarkably similar genomes and share closely related biochemical, physiological and pathological pathways. In this work we utilised the latest information on the mouse transcriptome as revealed by the RIKEN FANTOM2 project to identify novel human disease-related candidate genes. We define a new term "patholog" to mean a homolog of a human disease related gene encoding a product (transcript, anti-sense or protein) potentially relevant to disease. Rather than just focus on Mendelian inheritance, we applied the analysis to all potential pathologs regardless of their inheritance pattern. RESULTS: Bioinformatic analysis and human curation of 60,770 RIKEN full-length mouse cDNA clones produced 2,578 sequences that showed similarity (70-85% identity) to known human-disease genes. Using a newly developed biological information extraction and annotation tool (FACTS) in parallel with human expert analysis of 17,051 MEDLINE scientific abstracts we identified 182 novel potential pathologs. Of these, 36 were identified by computational tools only, 49 by human expert analysis only and 97 by both methods. These pathologs were related to neoplastic (53%), hereditary (24%), immunological (5%), cardio-vascular (4%), or other (14%), disorders. CONCLUSIONS: Large scale genome projects continue to produce a vast amount of data with potential application to the study of human disease. For this potential to be realised we need intelligent strategies for data categorisation and the ability to link sequence data with relevant literature. This paper demonstrates the power of combining human expert annotation with FACTS, a newly developed bioinformatics tool, to identify novel pathologs from within large-scale mouse transcript datasets. PMID- 15115541 TI - Influence of Tertiary paleoenvironmental changes on the diversification of South American mammals: a relaxed molecular clock study within xenarthrans. AB - BACKGROUND: Comparative genomic data among organisms allow the reconstruction of their phylogenies and evolutionary time scales. Molecular timings have been recently used to suggest that environmental global change have shaped the evolutionary history of diverse terrestrial organisms. Living xenarthrans (armadillos, anteaters and sloths) constitute an ideal model for studying the influence of past environmental changes on species diversification. Indeed, extant xenarthran species are relicts from an evolutionary radiation enhanced by their isolation in South America during the Tertiary era, a period for which major climate variations and tectonic events are relatively well documented. RESULTS: We applied a Bayesian approach to three nuclear genes in order to relax the molecular clock assumption while accounting for differences in evolutionary dynamics among genes and incorporating paleontological uncertainties. We obtained a molecular time scale for the evolution of extant xenarthrans and other placental mammals. Divergence time estimates provide substantial evidence for contemporaneous diversification events among independent xenarthran lineages. This correlated pattern of diversification might possibly relate to major environmental changes that occurred in South America during the Cenozoic. CONCLUSIONS: The observed synchronicity between planetary and biological events suggests that global change played a crucial role in shaping the evolutionary history of extant xenarthrans. Our findings open ways to test this hypothesis further in other South American mammalian endemics like hystricognath rodents, platyrrhine primates, and didelphid marsupials. PMID- 15115542 TI - Down-regulation of the M6P/IGF-II receptor increases cell proliferation and reduces apoptosis in neonatal rat cardiac myocytes. AB - BACKGROUND: The mannose 6-phosphate/insulin-like growth factor-II receptor (M6P/IGF2R) is a multi-functional protein that has been implicated in regulation of cell growth and apoptosis. Cardiac myocytes express relatively high levels of M6P/IGF2R, and cardiomyocyte apoptosis has been identified in a variety of cardiovascular disorders, such as myocardial infarction and heart failure. However, involvement of M6P/IGF2R in the pathogenesis of these conditions has not been determined. Thus, the objective of this study was to determine the role of M6P/IGF2R in regulation of cardiac myocyte growth and apoptosis. RESULTS: We down regulated the expression of M6P/IGF2R in neonatal rat cardiac myocytes and examined the effect on cell proliferation and apoptosis. Infection of neonatal cardiomyocytes with an adenovirus expressing a ribozyme targeted against the M6P/IGF2R significantly reduced the level of M6P/IGF2R mRNA, as determined by RT PCR and Ribonuclease Protection Assay (RPA). M6P-containing protein binding and endocytosis as well as the M6P/IGF2R-mediated internalization of 125I-IGF-II were lower in the ribozyme-treated cells than the control myocytes, indicating that the number of functional M6P/IGF2R in the ribozyme treated cells was reduced. Accordingly, a marked increase in cell proliferation and a reduced cell susceptibility to hypoxia- and TNF-induced apoptosis were observed in the ribozyme-treated cells. CONCLUSIONS: These findings suggest that M6P/IGF2R may play a role in regulation of cardiac myocyte growth and apoptosis. Down regulation of this gene in cardiac tissues might be a new approach to prevention of cell death or promotion of mitogenesis for certain heart diseases. PMID- 15115543 TI - A hybrid clustering approach to recognition of protein families in 114 microbial genomes. AB - BACKGROUND: Grouping proteins into sequence-based clusters is a fundamental step in many bioinformatic analyses (e.g., homology-based prediction of structure or function). Standard clustering methods such as single-linkage clustering capture a history of cluster topologies as a function of threshold, but in practice their usefulness is limited because unrelated sequences join clusters before biologically meaningful families are fully constituted, e.g. as the result of matches to so-called promiscuous domains. Use of the Markov Cluster algorithm avoids this non-specificity, but does not preserve topological or threshold information about protein families. RESULTS: We describe a hybrid approach to sequence-based clustering of proteins that combines the advantages of standard and Markov clustering. We have implemented this hybrid approach over a relational database environment, and describe its application to clustering a large subset of PDB, and to 328577 proteins from 114 fully sequenced microbial genomes. To demonstrate utility with difficult problems, we show that hybrid clustering allows us to constitute the paralogous family of ATP synthase F1 rotary motor subunits into a single, biologically interpretable hierarchical grouping that was not accessible using either single-linkage or Markov clustering alone. We describe validation of this method by hybrid clustering of PDB and mapping SCOP families and domains onto the resulting clusters. CONCLUSION: Hybrid (Markov followed by single-linkage) clustering combines the advantages of the Markov Cluster algorithm (avoidance of non-specific clusters resulting from matches to promiscuous domains) and single-linkage clustering (preservation of topological information as a function of threshold). Within the individual Markov clusters, single-linkage clustering is a more-precise instrument, discerning sub-clusters of biological relevance. Our hybrid approach thus provides a computationally efficient approach to the automated recognition of protein families for phylogenomic analysis. PMID- 15115544 TI - Identification of regions in multiple sequence alignments thermodynamically suitable for targeting by consensus oligonucleotides: application to HIV genome. AB - BACKGROUND: Computer programs for the generation of multiple sequence alignments such as "Clustal W" allow detection of regions that are most conserved among many sequence variants. However, even for regions that are equally conserved, their potential utility as hybridization targets varies. Mismatches in sequence variants are more disruptive in some duplexes than in others. Additionally, the propensity for self-interactions amongst oligonucleotides targeting conserved regions differs and the structure of target regions themselves can also influence hybridization efficiency. There is a need to develop software that will employ thermodynamic selection criteria for finding optimal hybridization targets in related sequences. RESULTS: A new scheme and new software for optimal detection of oligonucleotide hybridization targets common to families of aligned sequences is suggested and applied to aligned sequence variants of the complete HIV-1 genome. The scheme employs sequential filtering procedures with experimentally determined thermodynamic cut off points: 1) creation of a consensus sequence of RNA or DNA from aligned sequence variants with specification of the lengths of fragments to be used as oligonucleotide targets in the analyses; 2) selection of DNA oligonucleotides that have pairing potential, greater than a defined threshold, with all variants of aligned RNA sequences; 3) elimination of DNA oligonucleotides that have self-pairing potentials for intra- and inter-molecular interactions greater than defined thresholds. This scheme has been applied to the HIV-1 genome with experimentally determined thermodynamic cut off points. Theoretically optimal RNA target regions for consensus oligonucleotides were found. They can be further used for improvement of oligo-probe based HIV detection techniques. CONCLUSIONS: A selection scheme with thermodynamic thresholds and software is presented in this study. The package can be used for any purpose where there is a need to design optimal consensus oligonucleotides capable of interacting efficiently with hybridization targets common to families of aligned RNA or DNA sequences. Our thermodynamic approach can be helpful in designing consensus oligonucleotides with consistently high affinity to target variants in evolutionary related genes or genomes. PMID- 15115545 TI - Is an excretory urogram mandatory in patients with small to medium-sized renal and ureteric stones treated by extra corporeal shock wave lithotripsy? AB - BACKGROUND: An intravenous urogram (IVU) has traditionally been considered mandatory before treating renal and ureteric stones by extracorporeal shock wave lithotripsy (ESWL). This study was designed to see whether there is a difference in complications and the need for ancillary procedures in patients managed by ESWL for renal and ureteric calculi, according to preoperative imaging technique. METHODS: This retrospective study compared 133 patients undergoing ESWL from January 2001 to July 2002. Patients were divided into three groups according to the preoperative imaging technique used: i) IVU; ii) non-contrast enhanced helical computed tomography (UHCT); and iii) ultrasound (US) + X-ray kidney, ureter and bladder (KUB). The groups were matched in terms of age and gender, as well as location, side and size of stones. RESULTS: There was no statistically significantly difference for number of ESWL sessions, number of shock waves and use of ancillary procedures between the three groups. The stone-free rate was 98% for the IVU and UHCT groups, and 97% for the US + X-ray KUB group. CONCLUSIONS: The complication rate and need for ancillary procedures was comparable across the three groups. Patients imaged by UHCT or US + X-ray KUB prior to ESWL for uncomplicated renal and ureteric stones do not require IVU. PMID- 15115546 TI - Parasomnias and sleep disordered breathing in Caucasian and Hispanic children - the Tucson children's assessment of sleep apnea study. AB - BACKGROUND: Recent studies in children have demonstrated that frequent occurrence of parasomnias is related to increased sleep disruption, mental disorders, physical harm, sleep disordered breathing, and parental duress. Although there have been several cross-sectional and clinical studies of parasomnias in children, there have been no large, population-based studies using full polysomnography to examine the association between parasomnias and sleep disordered breathing. The Tucson Children's Assessment of Sleep Apnea study is a community-based cohort study designed to investigate the prevalence and correlates of objectively measured sleep disordered breathing (SDB) in pre adolescent children six to 11 years of age. This paper characterizes the relationships between parasomnias and SDB with its associated symptoms in these children. METHODS: Parents completed questionnaires pertaining to their child's sleep habits. Children had various physiological measurements completed and then were connected to the Compumedics PS-2 sleep recording system for full, unattended polysomnography in the home. A total of 480 unattended home polysomnograms were completed on a sample that was 50% female, 42.3% Hispanic, and 52.9% between the ages of six and eight years. RESULTS: Children with a Respiratory Disturbance Index of one or greater were more likely to have sleep walking (7.0% versus 2.5%, p < 0.02), sleep talking (18.3% versus 9.0%, p < 0.006), and enuresis (11.3% versus 6.3%, p < 0.08) than children with an Respiratory Disturbance Index of less than one. A higher prevalence of other sleep disturbances as well as learning problems was observed in children with parasomnia. Those with parasomnias associated with arousal were observed to have increased number of stage shifts. Small alterations in sleep architecture were found in those with enuresis. CONCLUSIONS: In this population-based cohort study, pre-adolescent school-aged children with SDB experienced more parasomnias than those without SDB. Parasomnias were associated with a higher prevalence of other sleep disturbances and learning problems. Clinical evaluation of children with parasomnias should include consideration of SDB. PMID- 15115548 TI - Pilfering for survival: how health workers use access to drugs as a coping strategy. AB - BACKGROUND: Coping strategies have, in some countries, become so prevalent that it has been widely assumed that the very notion of civil services ethos has completely - and possibly irreversibly - disappeared. This paper describes the importance and the nature of pilfering of drugs by health staff in Mozambique and Cape Verde, as perceived by health professionals from these countries. Their opinions provide pointers as to how to tackle these problems. METHODS: This study is based on a self-administered questionnaire addressed to a convenience sample of health workers in Mozambique and in Cape Verde. RESULTS: The study confirms that misuse of access to pharmaceuticals has become a key element in the coping strategies health personnel develop to deal with difficult living conditions. Different professional groups (mis)use their privileged access in different ways, but doctors diversify most. The study identifies the reasons given for misusing access to drugs, shows how the problem is perceived by the health workers, and discusses the implications for finding solutions to the problem.Our findings reflect, from the health workers themselves, a conflict between their self image of what it means to be an honest civil servant who wants to do a decent job, and the brute facts of life that make them betray that image. The manifest unease that this provokes is an important observation as such. CONCLUSION: Our findings suggest that, even in the difficult circumstances observed in many countries, behaviours that depart from traditional civil servant deontology have not been interiorised as a norm. This ambiguity indicates that interventions to mitigate the erosion of proper conduct would be welcome. The time to act is now, before small-scale individual coping grows into large-scale, well-organized crime. PMID- 15115549 TI - The role of wages in the migration of health care professionals from developing countries. AB - Several countries are increasingly relying on immigration as a means of coping with domestic shortages of health care professionals. This trend has led to concerns that in many of the source countries - especially within Africa - the outflow of health care professionals is adversely affecting the health care system. This paper examines the role of wages in the migration decision and discusses the likely effect of wage increases in source countries in slowing migration flows.This paper uses data on wage differentials in the health care sector between source country and receiving country (adjusted for purchasing power parity) to test the hypothesis that larger wage differentials lead to a larger supply of health care migrants. Differences in other important factors affecting migration are discussed and, where available, data are presented.There is little correlation between the supply of health care migrants and the size of the wage differential between source and destination country. In cases where data are available on other factors affecting migration, controlling for these factors does not affect the result.At current levels, wage differentials between source and destination country are so large that small increases in health care wages in source countries are unlikely to affect significantly the supply of health care migrants. The results suggest that non-wage instruments might be more effective in altering migration flows. PMID- 15115547 TI - A systematic review of the incidence of schizophrenia: the distribution of rates and the influence of sex, urbanicity, migrant status and methodology. AB - BACKGROUND: Understanding variations in the incidence of schizophrenia is a crucial step in unravelling the aetiology of this group of disorders. The aims of this review are to systematically identify studies related to the incidence of schizophrenia, to describe the key features of these studies, and to explore the distribution of rates derived from these studies. METHODS: Studies with original data related to the incidence of schizophrenia (published 1965-2001) were identified via searching electronic databases, reviewing citations and writing to authors. These studies were divided into core studies, migrant studies, cohort studies and studies based on Other Special Groups. Between- and within-study filters were applied in order to identify discrete rates. Cumulative plots of these rates were made and these distributions were compared when the underlying rates were sorted according to sex, urbanicity, migrant status and various methodological features. RESULTS: We identified 100 core studies, 24 migrant studies, 23 cohort studies and 14 studies based on Other Special Groups. These studies, which were drawn from 33 countries, generated a total of 1,458 rates. Based on discrete core data for persons (55 studies and 170 rates), the distribution of rates was asymmetric and had a median value (10%-90% quantile) of 15.2 (7.7-43.0) per 100,000. The distribution of rates was significantly higher in males compared to females; the male/female rate ratio median (10%-90% quantile) was 1.40 (0.9-2.4). Those studies conducted in urban versus mixed urban rural catchment areas generated significantly higher rate distributions. The distribution of rates in migrants was significantly higher compared to native born; the migrant/native-born rate ratio median (10%-90% quantile) was 4.6 (1.0 12.8). Apart from the finding that older studies reported higher rates, other study features were not associated with significantly different rate distributions (e.g. overall quality, methods related to case finding, diagnostic confirmation and criteria, the use of age-standardization and age range). CONCLUSIONS: There is a wealth of data available on the incidence of schizophrenia. The width and skew of the rate distribution, and the significant impact of sex, urbanicity and migrant status on these distributions, indicate substantial variations in the incidence of schizophrenia. PMID- 15115551 TI - Immunohistochemical localization of integrin alpha V beta 3 and osteopontin suggests that they do not interact during embryo implantation in ruminants. AB - BACKGROUND: It has been suggested that trophoblast attachment requires co expression of integrin alpha V beta 3 and its ligand osteopontin at the fetal maternal interface. Until now the expression patterns of integrin alpha V beta 3 and osteopontin in the pregnant bovine uterus were unknown. The objectives of this study were to localize integrin alpha V beta 3 and osteopontin in bovine and sheep endometrium during the periimplantation period and to compare the distribution patterns using antibodies that had not yet been tested in sheep. METHODS: Cell compartments within endometrial tissue sections were scored for immunohistochemical staining intensity and data were analyzed to determine the effects of day of pregnancy or cycle. RESULTS: In pregnant bovine endometrium, integrin alpha V beta 3 was detected in luminal epithelium, stroma, myometrium and smooth muscle. A strong band of immunoreactivity was observed in the subepithelial stroma of intercaruncular regions, but there was reduced reactivity in the caruncles and glands. Bovine trophoblast did not express integrin alpha V beta 3 at any stage of pregnancy. In ovine endometrium a different pattern of staining for integrin alpha V beta 3 was observed. Reactivity was not present in the luminal epithelium or trophoblast. There was strong staining of the deep glands and no reactivity in the superficial glands. Osteopontin distribution was similar for sheep and cattle. For both species, apical staining was present on the luminal epithelium and glands and on embryonic tissues. CONCLUSION: In ruminants, integrin alpha V beta 3 and osteopontin do not co-localize at the fetal-maternal interface indicating that these proteins could not interact to facilitate embryo attachment as has been proposed in other species. PMID- 15115552 TI - WHO global campaigns: A way forward in addressing public health importance of common neurological disorders. PMID- 15115550 TI - Origin of germ cells and formation of new primary follicles in adult human ovaries. AB - Recent reports indicate that functional mouse oocytes and sperm can be derived in vitro from somatic cell lines. We hypothesize that in adult human ovaries, mesenchymal cells in the tunica albuginea (TA) are bipotent progenitors with a commitment for both primitive granulosa and germ cells. We investigated ovaries of twelve adult women (mean age 32.8 +/- 4.1 SD, range 27-38 years) by single, double, and triple color immunohistochemistry. We show that cytokeratin (CK)+ mesenchymal cells in ovarian TA differentiate into surface epithelium (SE) cells by a mesenchymal-epithelial transition. Segments of SE directly associated with ovarian cortex are overgrown by TA, forming solid epithelial cords, which fragment into small (20 micron) epithelial nests descending into the lower ovarian cortex, before assembling with zona pellucida (ZP)+ oocytes. Germ cells can originate from SE cells which cover the TA. Small (10 micron) germ-like cells showing PS1 meiotically expressed oocyte carbohydrate protein are derived from SE cells via asymmetric division. They show nuclear MAPK immunoexpression, subsequently divide symmetrically, and enter adjacent cortical vessels. During vascular transport, the putative germ cells increase to oocyte size, and are picked-up by epithelial nests associated with the vessels. During follicle formation, extensions of granulosa cells enter the oocyte cytoplasm, forming a single paranuclear CK+ Balbiani body supplying all the mitochondria of the oocyte. In the ovarian medulla, occasional vessels show an accumulation of ZP+ oocytes (25-30 microns) or their remnants, suggesting that some oocytes degenerate. In contrast to males, adult human female gonads do not preserve germline type stem cells. This study expands our previous observations on the formation of germ cells in adult human ovaries. Differentiation of primitive granulosa and germ cells from the bipotent mesenchymal cell precursors of TA in adult human ovaries represents a most sophisticated adaptive mechanism created during the evolution of female reproduction. Our data indicate that the pool of primary follicles in adult human ovaries does not represent a static but a dynamic population of differentiating and regressing structures. An essential mission of such follicular turnover might be elimination of spontaneous or environmentally induced genetic alterations of oocytes in resting primary follicles. PMID- 15115553 TI - Protection of rat renal vitamin E levels by ischemic-preconditioning. AB - BACKGROUND: During renal transplantation, the kidney remains without blood flow for a period of time. The following reperfusion of this ischemic kidney causes functional and structural injury. Formation of oxygen-derived free radicals (OFR) and subsequent lipid peroxidation (LP) has been implicated as the causative factors of these injuries. Vitamin E is known to be the main endogenous antioxidant that stabilizes cell membranes by interfering with LP. The present study was designed to examine the role of ischemic-preconditioning (repeated brief periods of ischemia, IPC) in prevention of renal injury caused by ischemia reperfusion (IR) in rats. METHODS: IPC included sequential clamping of the right renal artery for 5 min and release of the clamp for another 5 min for a 3 cycles. IR was induced by 30 min ischemia followed by 10 min reperfusion. Four groups of male rats were used: Control, IPC, IR and IPC-IR. Vitamin E, an endogenous antioxidant and as an index of LP, was measured by HPLC and UV detection in renal venous plasma and tissue. Renal function was assessed by serum creatinine and BUN levels. Renal damage was assessed in sections stained with Haematoxylin and Eosin. RESULTS: In the IR group, there was a significant decrease in vitamin E in plasma and tissue compared to a control group (p,0.05). In the IPC-IR group, vitamin E concentration was significantly higher than in the IR group (p,0.01). The results showed that 30 min ischemia in the IR group significantly (p,0.05) reduced renal function demonstrated by an increase in serum creatinine levels as compared with the control group. These results in the IPC group also showed a significant difference with the IR group but no significant difference in serum BUN and creatinine between IR and IPC-IR group were detected. Histological evaluation showed no structural damage in the IPC group and an improvement in the IPC-IR group compared to IR alone. CONCLUSIONS: In this study, IPC preserved vitamin E levels, but it could not markedly improve renal function in the early phase (1-2 h) of reperfusion. IPC may be a useful method for antioxidant preservation in organ transplantation. PMID- 15115554 TI - Intracluster correlation coefficients in cluster randomized trials: empirical insights into how should they be reported. AB - BACKGROUND: Increasingly, researchers are recognizing that there are many situations where the use of a cluster randomized trial may be more appropriate than an individually randomized trial. Similarly, the need for appropriate standards of reporting of cluster trials is more widely acknowledged. METHODS: In this paper, we describe the results of a survey to inform the appropriate reporting of the intracluster correlation coefficient (ICC)--the statistical measure of the clustering effect associated with a cluster randomized trial. RESULTS: We identified three dimensions that should be considered when reporting an ICC--a description of the dataset (including characteristics of the outcome and the intervention), information on how the ICC was calculated, and information on the precision of the ICC. CONCLUSIONS: This paper demonstrates the development of a framework for the reporting of ICCs. If adopted into routine practice, it has the potential to facilitate the interpretation of the cluster trial being reported and should help the development of new trials in the area. PMID- 15115555 TI - In vitro adhesion and invasion inhibition of Shigella dysenteriae, Shigella flexneri and Shigella sonnei clinical strains by human milk proteins. AB - BACKGROUND: Shigella is the etiological agent of shigellosis, a disease responsible for more than 500,000 deaths of children per year, in developing countries. These pathogens colonize the intestinal colon, invade, spreading to the other enterocytes. Breastfeeding plays a very important role in protecting infants from intestinal infections. Amongst milk compounds, glycosylated proteins prevent the adhesion of many enteropathogens in vitro. The aim of this work was to determine the effect of human milk proteins on the colonization potential of Shigella dysenteriae, S. flexneri and S. sonnei. To fulfill this purpose, pooled milk samples from five donors, were fractionated by gel filtration and affinity chromatography. Using tissue culture, the milk fractions obtained were tested in Shigella adhesion and invasion assays. RESULTS: Our revealed showed that both adhesion and invasion of Shigella species were inhibited by low concentration of secretory immunoglobulin A, lactoferrin and free secretory component. This work also showed that, these proteins bind to superficial and whole-cell Shigella proteins. CONCLUSIONS: Our findings suggest that human milk may act inhibiting adhesion and, consequently, invasion of Shigella, thereafter preventing shigellosis in infants. PMID- 15115556 TI - Forgoing life-sustaining treatments in children: a comparison between Northern and Southern European pediatric intensive care units. AB - OBJECTIVES: This study was conducted to determine how the decision-making process to forgo life support differs between southern and northern European pediatric intensive care units. DESIGN: Multiple-center, prospective study. SETTING: Thirty nine pediatric intensive care units: 12 from northern Europe and 27 from southern Europe. PATIENTS: All consecutive deaths were recorded over a 4-month period. Group 1 and group 2 included patients who died in northern and southern pediatric intensive care units, respectively. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Three hundred fifty children were enrolled, 68 in group 1 and 282 in group 2. The decision to forgo life-sustaining treatment was made in 116 children (group 1, n = 32; group 2, n = 84). In both groups, the decision was discussed by caregivers during a formal meeting. The decision to forgo life-sustaining treatment was more often made in northern countries than in southern ones (47% vs. 30%, p =.02). Parents were informed of this decision in 95% of cases in group 1 vs. 68% in group 2 (p =.01). In both groups, the final decision was made by the medical staff. Parents' contributions to the decision-making process did not differ between the two groups according to the practitioners' opinion. The decision was documented in the medical charts in 100% of the cases in group 1 and in 51% of the cases in group 2 (p =.0001). CONCLUSIONS: The decision-making process appears to be similar between northern and southern European countries. The respective contributions of the parents and the medical staff in the final decision itself seem to be identical between northern and southern countries. However, in northern European countries, the level of parents' information about the decision-making process appears higher and the decision is more often documented in the medical chart. PMID- 15115557 TI - A descriptive study of children dying in the pediatric intensive care unit after withdrawal of life-sustaining treatment. AB - OBJECTIVE: To examine physiologic and therapeutic changes following withdrawal of life-sustaining treatment in children. DESIGN: Retrospective chart review. SETTING: University-affiliated tertiary care pediatric hospital. PATIENTS: All patients who had life-sustaining treatment withdrawn over a 5-yr period. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: A total of 125 charts were examined to obtain 50 in which the terminal event preceding death was withdrawal of life-sustaining treatment. Data are expressed as median (1st, 3rd quartiles). Median hospital stay before death was 20 days (1st and 3rd quartiles, 8 and 30). Median time from decision to withdraw life-sustaining treatment to actual withdrawal was 30 mins (1st and 3rd quartiles, 10 and 180). All interventions were simultaneously discontinued in 80% of patients with mechanical ventilation followed by vasopressors being most common. No patients had stepwise reduction in ventilator rate before discontinuing the mechanical ventilation. Devices were rarely removed from patients including endotracheal tubes. Time from withdrawal of life-sustaining treatment to death was 15 mins (5, 30); only seven patients took >60 mins to die. Multivariable analysis (Kruskal-Wallis test) of various factors revealed simultaneous withdrawal of life-sustaining treatment, female gender, and not having received renal therapy as hastening death. CONCLUSIONS: Forgoing life-sustaining treatment in a small cohort of children at a single institution follows a pattern: Most cases occur after prolonged intensive care unit stays, withdrawal of treatment occurs almost immediately after the decision to withdraw, most treatments are withdrawn simultaneously rather than sequentially, and most patients die within minutes of life-sustaining treatment cessation. This is the first pediatric study to report the time to death after withdrawal of life-sustaining treatment and factors associated with shorter time to death in children. PMID- 15115558 TI - Sleep monitoring in children during neuromuscular blockade in the pediatric intensive care unit: a pilot study. AB - BACKGROUND: Sleep is an important physiologic process that is known to be disrupted in the intensive care unit. Nevertheless, there is little information on how intensive care unit admission affects sleep in children. Because laryngotracheoplasty is elective but entails 5-7 days of neuromuscular blockade following surgery, children undergoing this procedure present a unique opportunity to analyze sleep during neuromuscular blockade apart from confounding variables resulting from critical illness. OBJECTIVE: To determine the feasibility of using polysomnography to assess sleep patterns in children during neuromuscular blockade. METHODS: Polysomnography recordings were obtained continuously for 4 days (96 hrs) in two children following laryngotracheoplasty. Medication administration (neuromuscular blockades, sedatives) and time of suctioning were also recorded. RESULTS: Both subjects had documented sleep. However, the proportion of time in each stage was markedly different from developmental norms, and a greater proportion of sleep occurred during the day. Furthermore, there was substantial day-night and day-to-day variability. Some rebound of consolidated sleep appeared by day 4. Sedative use varied considerably. However, neither bolus sedation administration nor endotracheal suctioning appeared to affect sleep. Few monitoring difficulties were encountered. CONCLUSIONS: Sleep can be monitored with minimal difficulty in children undergoing neuromuscular blockade in the pediatric intensive care unit. Sleep occurred throughout the day, and there was considerable fragmentation. To fully assess sleep in the intensive care unit, monitoring needs to be continuous over several days, rather than only at night or for < or =24 hrs. Further research is needed in the area to determine typical sleep patterns in children undergoing neuromuscular blockade. PMID- 15115559 TI - Loss of consciousness: when to perform computed tomography? AB - OBJECTIVE: To determine the diagnostic value of physical examination (including neurologic exam) for positive computed tomography scan findings in children with closed head injury, Glasgow Coma Scale score 13-15 in the emergency department, and loss of consciousness or amnesia. DESIGN: Prospective descriptive study. SETTING: A large, tertiary, pediatric trauma center in San Diego County. PATIENTS: Children ages 2-16 with an isolated closed head injury, history of loss of consciousness or amnesia, and Glasgow Coma Scale 13-15 who were referred for pediatric trauma evaluation and received a head computed tomography as part of this evaluation. INTERVENTIONS: A standardized physical examination including skull/scalp exam, pupils, tympanic membrane, and brief neurologic exam was documented on each patient. MEASUREMENTS AND MAIN RESULTS: Subjects age 2-16 being evaluated by the pediatric trauma team for closed head injury with loss of consciousness or amnesia and Glasgow Coma Scale 13-15 received a standardized physical exam, noncontrast head computed tomography scan, and follow-up telephone call at 4-6 wks. Outcome variables include intracranial injury visualized on computed tomography scan and need for neurosurgical intervention. Ninety-eight subjects were enrolled in the study over a 1-yr period. Computed tomography scans revealed evidence of intracranial injury in 13 of 98 subjects (13%). Normal examination increased the probability of a normal computed tomography scan from.87 pretest to.90 posttest. Four of 38 subjects with normal examination were noted to have evidence of intracranial injury on computed tomography. These four subjects did not require neurosurgical intervention. Two of 98 subjects underwent neurosurgical procedures. One intracranial pressure monitor was placed for decreasing level of consciousness. One subject underwent surgical elevation of a depressed skull fracture. CONCLUSIONS: Detailed clinical examination is of no diagnostic value in detecting intracranial injuries found on head computed tomography scan. Patients with observed loss of consciousness or amnesia and Glasgow Coma Scale 13-15 should have a head computed tomography scan as part of their evaluation to avoid missing an intracranial injury. PMID- 15115560 TI - Phosphodiesterase-3 inhibition prevents the increase in pulmonary vascular resistance following inhaled nitric oxide withdrawal in lambs. AB - OBJECTIVES: To determine the effects of inhaled nitric oxide on endogenous cyclic adenosine monophosphate in the intact lamb, and to determine the potential role of cyclic adenosine monophosphate in the rebound pulmonary hypertension associated with nitric oxide withdrawal. DESIGN: Prospective, placebo-controlled experimental study. SETTING: University-based basic science research laboratory. SUBJECTS: One-month-old lambs. INTERVENTIONS: Six 1-month-old control lambs, and 6 milrinone- (phosphodiesterase-3 inhibitor) treated lambs, were mechanically ventilated. Inhaled nitric oxide (40 ppm) was administered for 24 hrs and then acutely withdrawn. Sequential peripheral lung biopsies were obtained before, during, and 2 hrs after withdrawing inhaled nitric oxide therapy. MEASUREMENTS AND MAIN RESULTS: In control lambs, initiation of nitric oxide decreased left pulmonary vascular resistance by 29.6%, and withdrawal rapidly increased pulmonary vascular resistance by 77.1% (p <.05). Lung tissue cyclic adenosine monophosphate concentrations decreased by 25.3% during nitric oxide therapy (p <.05). In milrinone-treated lambs, nitric oxide decreased pulmonary vascular resistance by 26.6% (p <.05), but pulmonary vascular resistance was unchanged after acute withdrawal. Lung tissue cyclic adenosine monophosphate concentrations were preserved during nitric oxide therapy. CONCLUSIONS: Inhaled nitric oxide produces potent pulmonary vasodilation by activating soluble guanylate cyclase and increasing smooth muscle cell concentrations of guanosine-3',5'-cyclic monophosphate. However, alterations in endogenous nitric oxide/guanosine-3',5' cyclic monophosphate during inhaled nitric oxide have been implicated in the clinically significant increases in pulmonary vascular resistance noted upon its acute withdrawal. Previous in vitro data suggest that exogenous nitric oxide/guanosine-3',5'-cyclic monophosphate can also alter cyclic adenosine monophosphate concentrations via their effect on cyclic adenosine monophosphate production and metabolism. The current in vivo study demonstrates that lung tissue cyclic adenosine monophosphate concentrations are decreased during inhaled nitric oxide and suggests a role for decreased cyclic adenosine monophosphate in the rebound pulmonary hypertension noted upon inhaled nitric oxide withdrawal. Milrinone may be a useful adjunct therapy during inhaled nitric oxide to preserve cyclic adenosine monophosphate concentrations and prevent rebound pulmonary hypertension. PMID- 15115561 TI - Defining acidosis in postoperative cardiac patients using Stewart's method of strong ion difference. AB - OBJECTIVE: To define the true incidence and nature of acidosis in pediatric patients postcardiac surgery, using Stewart's direct method of measuring strong ion difference. We also wished to compare the ability of standard indirect methods (base deficit, lactate, anion gap, and corrected anion gap) to accurately predict tissue acidosis. DESIGN: A single-center prospective observational study. SETTING: A pediatric intensive care unit in a tertiary referral center. PATIENTS: Pediatric patients who had undergone cardiac surgery were studied in the immediate postoperative period. Patients who had undergone both open and closed cardiac surgery were included. INTERVENTIONS: Routine arterial blood gas analysis and laboratory electrolyte measurements were made in patients immediately on admission to the pediatric intensive care unit (PICU) after cardiac surgery and each morning until discharge from the PICU. MEASUREMENTS AND MAIN RESULTS: Figge's equations were used to calculate strong ion difference and total tissue acids (unmeasured acids and lactate). These direct methods then were compared to indirect measurements: base deficit, lactate anion gap, and anion gap corrected for albumin. We collected 150 samples from 44 patients. Tissue acidosis occurred overall in 60 of 150 samples. This was due to raised unmeasured acids alone in 44 of 60 (73.3%), raised lactate alone in six of 60 (10%), and a combination of the two in ten of 60 (16.6%). Hyperchloremia occurred in 19 of 150 samples overall and 12 of 25 (48%) samples immediately after cardiopulmonary bypass. Measured base deficit showed a poor correlation with true tissue acidosis (r = -.48, p <.001) and the worst discriminatory ability (area under the curve, 0.72; 0.62 0.82). Anion gap corrected for albumin had the best correlation (r =.95, p <.001) and highest area under the curve (0.90; 0.85-0.95). CONCLUSIONS: Metabolic acidosis occurs frequently postcardiac surgery and is largely due to raised unmeasured acids and less commonly raised lactate. Hyperchloremia is common, particularly after cardiopulmonary bypass. Base deficit correlates poorly with true tissue acidosis, and corrected anion gap offers the most accurate bedside alternative to Stewart's method of tissue acid calculation. PMID- 15115562 TI - The use of recombinant coagulation factor VIIa in uncontrolled postoperative bleeding in children undergoing cardiac surgery with cardiopulmonary bypass. AB - OBJECTIVE: To assess the hemostatic efficacy of recombinant coagulation factor VIIa (rFVIIa) in the management of uncontrolled bleeding in postcardiac surgery with cardiopulmonary bypass in children. DESIGN: An open-label study. SETTING: A postoperative intensive care unit. PATIENTS: Eight consecutive pediatric patients with excessive bleeding after cardiac surgery with cardiopulmonary bypass that met the criteria for reexploration and did not respond to optimal transfusions of platelets and fresh frozen plasma. INTERVENTIONS: rFVIIa 30 microg/kg was given as a bolus injection. A higher dose of 60 microg/kg was used if a patient had preoperative coagulopathy, preoperative multiple-organ failure, or indications that required an emergency operation. The same dose was repeated 15 mins after the previous injection if the bleeding had not decreased. If the bleeding had decreased but still exceeded 10 mL/hr for body weight 5 kg, the same dose was repeated 2 hrs after the previous injection. A maximum of four doses could be given before rFVIIa was considered ineffective and a reexploration was needed. MEASUREMENTS AND MAIN RESULTS: Postoperative blood loss was estimated from the volume of chest tube drainage. rFVIIa successfully controlled bleeding and prevented reexploration in all seven patients who received treatment according to the protocol. One patient who received only one dose of rFVIIa required reexploration because a second dose was not available. No adverse events related to rFVIIa were seen. CONCLUSIONS: rFVIIa may be useful in preventing reexploration in uncontrolled postoperative bleeding in children undergoing cardiac surgery with cardiopulmonary bypass. Randomized, placebo-controlled studies are needed to confirm the safety and efficacy of rFVIIa in this clinical setting. PMID- 15115563 TI - The use of telemedicine to provide pediatric critical care consultations to pediatric trauma patients admitted to a remote trauma intensive care unit: a preliminary report. AB - OBJECTIVE: Injured pediatric patients in remote communities are often cared for at trauma centers that may be underserved with respect to pediatric specialty services. The objective of this study is to describe a pilot telemedicine project that allows a remote trauma center's adult intensive care unit to obtain nontrauma, nonsurgical-related pediatric critical care consultations for acutely injured children. DESIGN: Nonconcurrent cohort design. SETTING: A remote, level II trauma center's shock-trauma intensive care unit and a tertiary care children's hospital pediatric intensive care unit. PATIENTS: Analyses were conducted on cohorts of pediatric trauma patients (<16 yrs) consecutively admitted to the remote adult intensive care unit, including historical control patients and patients who received and did not receive telemedicine consultations. INTERVENTIONS: Telemedicine consultations were obtained at the discretion of the remote intensive care unit provider for nontrauma, nonsurgical medical issues. MEASUREMENTS AND RESULTS: The Injury Severity Score and Trauma and Injury Severity Score were used to assess severity of injury and predicted mortality rates, respectively, for the patient cohorts. Parental and provider satisfaction with the telemedicine consultations was also described. Thirty-nine consultations were conducted on 17 patients from the 97 pediatric patients admitted during the 2-yr study. Patients who received consultations were younger (5.5 yrs vs. 13.3 yrs, p <.01) and were more severely injured (mean Injury Severity Score = 18.3 vs. 14.7, p =.07). Severity-adjusted mortality rates were consistent with Trauma and Injury Severity Score expectations. Satisfaction surveys suggested a high level of provider and parental satisfaction. CONCLUSIONS: Our report of a trauma intensive care unit based pediatric critical care telemedicine program demonstrates that telemedicine consultations to a remote intensive care unit are feasible and suggests a high level of satisfaction among providers and parents. PMID- 15115564 TI - Cerebral autoregulation in pediatric traumatic brain injury. AB - OBJECTIVE: The aims of this study were to document the incidence of impaired cerebral autoregulation in children with traumatic brain injury using transcranial Doppler ultrasonography and to examine the relationship between autoregulatory capacity and outcome in children following traumatic brain injury. DESIGN: Prospective cohort study. SETTING: Harborview Medical Center (level I pediatric trauma center) in Washington state. PATIENTS: Thirty-six children <15 yrs old with traumatic brain injury: Glasgow Coma Scale score <9 (n = 12, group 1), Glasgow Coma Scale score 9-12 (n = 12, group 2), and Glasgow Coma Scale score 13-15 (n = 12, group 3). INTERVENTIONS: Cerebral autoregulation testing was conducted during extracranial surgery. Mean middle cerebral artery flow velocities were measured using transcranial Doppler as mean arterial pressure was increased to whichever variable was greater: 20% above baseline or a set value (80 mm Hg for <9 yrs and 90 mm Hg for 9-14 yrs). Autoregulatory capacity was quantified by the Autoregulatory Index. Autoregulatory Index <0.4 was considered impaired cerebral autoregulation. Discharge outcome using the Glasgow Outcome Scale score was considered good if the Glasgow Outcome Scale score was > or =4. MEASUREMENTS AND MAIN RESULTS: Twenty-four (67%) of 36 children had an Autoregulatory Index > or =0.4. The incidence of impaired cerebral autoregulation was 42% (five of 12) in group 1, 42% (five of 12) in group 2, and 17% (two of 12) in group 3. Ten (42%) of the 24 children with intact cerebral autoregulation had a good outcome compared with only one of 12 (8%) children with impaired cerebral autoregulation (p =.04). Six of 12 (50%) children with impaired cerebral autoregulation had hyperemia compared with one of 24 (4%) children with intact cerebral autoregulation (p <.01). Hyperemia was associated with poor outcome (p =.01). CONCLUSIONS: The incidence of impaired cerebral autoregulation was greatest following moderate to severe traumatic brain injury. Impaired cerebral autoregulation was associated with poor outcome. Hyperemia was associated with impaired cerebral autoregulation and poor outcome. PMID- 15115565 TI - Low cerebral blood flow resistance in nonventilated preterm infants predicts poor neurologic outcome. AB - OBJECTIVES: To examine whether cerebral blood flow variables during the first critical day of life can predict the 1-yr neurologic outcome in ventilated and nonventilated preterm infants. DESIGN: Prospective follow-up study. SETTING: Neonatal intensive care unit of university central hospital. PATIENTS: Forty-nine preterm infants <33 wks of gestation. INTERVENTIONS: Doppler ultrasound investigations of the brain circulation, heart rate, and systemic blood pressure were performed in ventilated (n = 35) and nonventilated (n = 14) preterm infants during the first day of life. The neurologic development was evaluated using Griffith's subscales at 12 months of corrected age. MEASUREMENTS AND MAIN RESULTS: Cerebral blood flow velocity measurements were obtained from the anterior cerebral artery and internal carotid artery. Cerebral blood flow, cerebral blood flow resistance, and cerebral perfusion pressure subsequently were derived. These derived cerebral perfusion variables were associated with the sum of Griffith's developmental scales (p <.02). However, the slopes of regression lines between cerebral blood flow or cerebral blood flow resistance and the sum of Griffith's psychomotor developmental scales tended to be different in the ventilated and nonventilated infants (p =.06, p =.003, respectively). The correlations between these variables and the sum of Griffith's psychomotor developmental scales were significant only in nonventilated preterm infants (r =.69, p =.007, and r = -.85, p =.001, respectively). CONCLUSIONS: Our data suggest that lowered cerebral blood flow resistance reflecting lowered cerebral blood flow during early circulatory transition is associated with adverse outcome in nonventilated preterm infants, but no connection in ventilated infants was found. PMID- 15115566 TI - Comparison of solute clearance in three modes of continuous renal replacement therapy. AB - OBJECTIVES: To compare the clearances of low molecular weight molecules using three modalities of continuous renal replacement therapy (CRRT) at the low blood flow rates typically used in pediatric patients. DESIGN: A controlled, in vitro laboratory study. SETTING: Research laboratory of a health sciences university. SUBJECTS: AN69 dialysis hemofilter. INTERVENTIONS: CRRT was performed using a 0.6 m(2) AN69 hemofilter. Human whole blood and plasma were combined to achieve a hematocrit of approximately 30%. Urea and creatinine were added to obtain concentrations of approximately 54 mmol/L of blood urea nitrogen and 1770 micromol/L of creatinine. Clearance data for urea and creatinine at a blood flow rate of 60 mL/min were generated using predilution continuous venovenous hemofiltration (CVVH), postdilution CVVH, and continuous venovenous hemodialysis (CVVHD). MEASUREMENTS AND MAIN RESULTS: Clearance of all three modalities was compared at a replacement solution (CVVH) or dialysate (CVVHD) flow rate of 16.7% of the blood flow rate. Both postdilution CVVH and CVVHD had a urea clearance of 11.3 mL/min, which was 15% greater than the 9.8 mL/min urea clearance of predilution CVVH (p <.005). Creatinine clearance with postdilution CVVH (10.7 mL/min) was 15% greater than the 9.0 mL/min clearance produced by predilution CVVH (p < 0.01). Predilution CVVH and CVVHD were compared at a flow rate of either replacement solution (CVVH) or dialysate (CVVHD) of 33% and 50% of the blood flow rate. Postdilution CVVH was not performed at high ultrafiltration rates due to the potential problem of hemoconcentration. CVVHD clearances of urea and creatinine were statistically superior to predilution CVVH at both flow rates. CONCLUSIONS: CVVHD was superior to predilution CVVH for clearance of urea and creatinine. Postdilution CVVH and CVVHD gave nearly equivalent clearances. At the low blood flow rates used in pediatric patients, which raise concerns about high ultrafiltration during postdilution CVVH causing excessive hemoconcentration and filter clotting, CVVHD appears to be the optimal modality for maximizing clearance of small solutes during CRRT. PMID- 15115567 TI - Jading in the pediatric intensive care unit: Implications for healthcare providers of medically complex children. AB - OBJECTIVE: To discuss the phenomenon of jading within the context of the pediatric intensive care unit. DESIGN: Drawing from their experience, the authors describe and then discuss a clinical scenario readily recognizable by pediatric intensive care unit practitioners: a child whose care requires the expenditure of a large amount of energy and resources, provides seemingly little reward, and leads to jading of the PICU staff. CONCLUSION: Jading describes a process of exhaustion whereby apathy, cynicism, and callousness replace the drive to be responsive, to make a difference, and to care. The issue of jading has become an increasing area of concern in the pediatric intensive care unit, due in part to recurring, prolonged admissions, combined with the perception, at times, that continued medical care is fruitless. With a better understanding of the phenomenon of jading, and by reconsidering their own responses, pediatric intensive care unit practitioners can avoid becoming jaded. PMID- 15115568 TI - Amino acid clearance during acute metabolic decompensation in maple syrup urine disease treated with continuous venovenous hemodialysis with filtration. AB - OBJECTIVE: Assessment of amino acid clearances by continuous venovenous hemodialysis with filtration in treatment of a metabolic decompensation in acute maple syrup urine disease. DESIGN: Single patient assessment. SETTING: Pediatric intensive care unit. PATIENTS: A 10-yr-old male with known maple syrup urine disease (branched chain alpha-ketoacid dehydrogenase deficiency) with metabolic decompensation due to an acute viral illness, characterized by altered mental status, progressive obtundation, and severe acidosis. INTERVENTIONS: Continuous venovenous hemodialysis with filtration. MEASUREMENTS AND MAIN RESULTS: Continuous venovenous hemodialysis with filtration was instituted with both filtration (500 mL/m(2)/hr) and dialysis (1000 mL/m(2)/hr) utilized, allowing rapid correction of systemic ketoacidosis while providing amino acid clearance. Amino acid clearance was measured at initiation and at 24 hrs into therapy. The procedure was well tolerated, with near normal mental status within 12 hrs and resumption of enteral feedings. During the 24-hr period of continuous venovenous hemodialysis with filtration, serum leucine levels fell from 2352 to 381 micromoles/L, isoleucine fell from 626 to 164, and valine fell from 1117 to 228. Leucine, isoleucine, and valine clearance rates averaged 13.1, 12.8, and 13.2 mL/min, respectively, and were constant during the 24 hrs of treatment. Clearance of other amino acids during this period did not vary significantly between cationic, anionic, neutral, or hydrophobic amino acids. CONCLUSIONS: Continuous venovenous hemodialysis with filtration provides an effective therapeutic alternative to intermittent hemodialysis during acute metabolic decompensation in maple syrup urine disease. PMID- 15115569 TI - Adrenoceptor agonist poisoning after accidental oral ingestion of brimonidine eye drops. AB - OBJECTIVE: Brimonidine tartrate is a relatively selective alpha(2) adrenic agonist that lowers elevated intraocular pressure. A 2-yr-old boy presented with severe cardiorespiratory symptoms of systemic alpha(2) adrenergic intoxication after accidentally ingesting 2 mL of brimonidine ophthalmic solution (0.2%) orally. At 20 mins after ingestion, he became acutely pale and lethargic, with shallow infrequent respirations. The symptoms resolved completely within the next 10 hrs. METHODS: The pharmacokinetics of brimonidine in plasma and urine were analyzed using gas chromatography and mass spectrometry. RESULTS: Maximum plasma concentrations were 40 ng/mL 5 hrs after ingestion. The plasma elimination half life value was 2.7 hrs. Elimination via urine was calculated as having a terminal half-life value of 3.2 hrs. CONCLUSIONS: This case illustrates the ability of brimonidine to gain rapid access to the central nervous system. This first passage is followed by a redistribution phase with rising plasma concentrations. Children who accidentally ingest brimonidine orally should be admitted to a pediatric intensive care unit. PMID- 15115570 TI - Polyarteritis nodosa with central nervous system involvement mimicking meningoencephalitis. AB - OBJECTIVE: To describe a patient who had polyarteritis nodosa with central nervous system involvement mimicking infectious meningoencephalitis. DESIGN: Case report. SETTING: Pediatric intensive care unit of a university hospital. PATIENT: A 9-yr-old boy with prolonged fever, headache, decreased level of consciousness, neck stiffness, and papilledema. RESULTS: Cerebrospinal fluid examination showed pleocytosis and a high protein level. After neurologic deterioration resulted from the initial treatment with antibiotic, the combination of clinical and laboratory findings with neuroradiologic features led to suspected systemic vasculitis. The patient was treated subsequently with corticosteroid, which resulted in great improvement. Biopsy of a skin lesion confirmed the diagnosis of polyarteritis nodosa. CONCLUSIONS: Critical care physicians must recognize neurologic manifestation patterns of systemic vasculitides because appropriate diagnosis and therapy result in significantly improved morbidity and mortality. PMID- 15115571 TI - The Fourth World Congress on pediatric intensive care. PMID- 15115573 TI - The quality of care provided to vulnerable older community-based patients with urinary incontinence. AB - OBJECTIVE: The objective of this study was to assess the quality of care provided to vulnerable older community-based patients with urinary incontinence (UI). DESIGN: We conducted an observational study using medical record review and patient (or proxy) interview. PARTICIPANTS: We studied 372 randomly selected community-dwelling older patients enrolled in two senior managed care plans identified by interview to be at increased risk for functional decline or death. MEASUREMENTS: Percentage of quality indicators passed for patients with UI. RESULTS: Thirty-six percent of the patients reported having UI, and during a 13 month period, 32 (7%) presented to their physician with new or worsening UI. Analysis of medical records for these 32 patients revealed that characteristics of voiding were documented for 75% of the patients with new or worsening UI, but importance of the problem, toileting function, and prior treatment were rarely addressed. Pelvic examination was performed for 20% of female patients and a rectal examination for 42% of men. Only 38% had a urinalysis performed and 16% had a postvoid residual. Drug treatment was prescribed for 50% of the patients but behavioral intervention for only 13%. Compared with patients seen by primary care providers alone, patients seen in consultative care received more comprehensive evaluation and treatment of UI. CONCLUSION: Quality of care for UI provided to vulnerable older patients, particularly by primary care providers alone, is inadequate. Despite the proven effectiveness of patient-dependent behavioral treatments, physicians rarely prescribe these interventions for UI. PMID- 15115574 TI - End-of-life care in U.S. nursing homes: a review of the evidence. AB - OBJECTIVES: The purpose of this study was to systematically review the empiric evidence on end-of-life care in nursing homes in the United States The guiding research question for this review was what is the state of research evidence in end-of-life care in long-term care? DESIGN: We conducted a systematic review of the literature. DATA: The review was limited to published and indexed research in peer-reviewed journals in five major databases between 1995 and October 2002. RESULTS: The initial search yielded a total of 395 articles. The search was narrowed, focusing on nursing homes in the United States and empiric research. The result was 43 articles related to research in end-of-life care in American nursing homes. It was categorized into eight foci: prognosis, pain, hospice, hospitalization, advanced care planning, communication, family perceptions, and miscellaneous. CONCLUSION: There is a dearth of research published in end-of-life care in the nursing home setting. What is available is primarily descriptive. The empiric research only documents poor end-of-life care in U.S. nursing homes. Empiric evidence has grown in this area, but there is now a need for research of creative and innovative solutions aimed at improving the quality of end-of-life care in this setting. PMID- 15115575 TI - Eye care in the skilled nursing facility: a pilot study of prevalence and treatment patterns of glaucoma. AB - OBJECTIVE: The objective of this study was to determine the rates of ophthalmic examinations for glaucoma, prevalence rates of glaucoma, ongoing evaluation (follow-up) rates and rates of treatment for a population of residents in a skilled nursing facility. DESIGN: We conducted a retrospective evaluation and chart review of glaucoma-related ophthalmology services. SETTING: This study was conducted in a skilled nursing facility located in a large metropolitan area located in the Midwest. PARTICIPANTS: We studied all nursing home residents of the facility on October 1, 2002 (n = 160). METHODS: We conducted a retrospective evaluation and chart review of glaucoma-related ophthalmology services for 160 patients. The medical records used for review included admission records, physician history and physical records, hospital notes, nursing assessments, consultation notes, and medication reviews (including medications administered during hospital stays). Minimum data set (MDS) data and individual patient interviews were used to supplement and verify chart abstraction findings. RESULTS: Eighty-three residents (52%) had evidence of assessment for glaucoma. Thirty-three of these residents (40%) had documentation of a diagnosis of glaucoma; 25 (76%) had current treatment orders for a topical ophthalmic agent. Nine patients were using combination therapy; four used topical and oral beta adrenergic-blocking agents. CONCLUSIONS: Visual impairment remains a serious problem for nursing facility residents. Assessment of visual abilities is infrequent or nonexistent. Education for nursing home personnel, discussion and activation among nursing home thought leaders, and guidelines for the evaluation and management of glaucoma in this care environment are needed. PMID- 15115576 TI - Effect of high climate temperature on the behavioral and psychological symptoms of dementia. AB - OBJECTIVE: The objective of this study was to investigate if high climate temperature could have a negative effect on behavioral and psychological symptoms of dementia (BPSD). DESIGN: We conducted a retrospective study. SETTING AND PATIENTS: Six patients discharged from the "Richiedei" Alzheimer Rehabilitation Unit-Italy during a period of very high climate temperature (June 14-21, 2002, 30.4 degrees C [86.7 degrees F]) are defined as the case-group. Sociodemographic, clinical, and therapeutic characteristics are compared with those of patients discharged during the previous 5 weeks (I control-group, n = 10) and the following 5 weeks (II control-group, n = 9). METHODS: On admission and on discharge, a multidimensional evaluation was performed assessing mental and functional status, BPSD (Neuropsychiatric Inventory, NPI), somatic health status, and pharmacotherapy. Changes in BPSD severity was computed as the difference in NPI total score (Delta-NPI) between the last and the first week of stay. Atmospheric temperature was also considered. RESULTS: The three groups did not differ with respect to sociodemographic, clinical, and therapeutic characteristics. On admission, frequency and severity of BPSD were similar among groups; whereas on discharge, the case-group had an increase in NPI scores (NPI during the first week [median], 13 in the case-group vs. 21 in the I control group vs. 20.5 in the II one; during the last week: 24.5 vs. 9.5 vs. 15, respectively). CONCLUSION: High climate temperature could have a negative impact on behavior in demented patients. To explain the possible mechanisms of the negative impact of high climate temperature on behavior, two hypotheses are proposed. PMID- 15115577 TI - Psychosis in nursing home patients with Parkinson's disease. AB - Psychosis in patients with Parkinson's disease (PD) is a common and important problem. It is a frequent cause of nursing home placement and is associated with a high mortality. The cause of psychosis in PD is usually multi-factorial but often involves the agents used to treat the motor symptoms of the disease. Treatment strategies should include ruling out other causes of cognitive decline, reducing anti-PD medications, and judicious use of atypical neuroleptic medications. Cholinesterase inhibitors can also be useful as an adjunctive agent in patients with psychosis and cognitive decline. Careful management of PD patients with psychosis can often result in good control of hallucinations and delusions without significant decline in motor function. PMID- 15115578 TI - Can we prevent aspiration pneumonia in the nursing home? AB - Aspiration pneumonia is a significant cause of morbidity, hospitalization, and mortality in the nursing home population. Patients who aspirate have three times higher mortality than patients who do not aspirate. We discuss the factors known to increase the risk of aspiration and its consequences, and recognize some of the preventive measures for aspiration pneumonia. We suggest approaches to decrease the risk of this very prevalent syndrome. PMID- 15115579 TI - Nursing documentation of telephone communication with physicians in community nursing homes. AB - OBJECTIVE: The objective of this study was to describe the nursing documentation of telephone communication with physicians in community nursing homes. DESIGN: We conducted a prospective observational study evaluating nursing documentation of all telephone calls to one physician group. SETTING: We studied two proprietary community nursing homes in Baltimore, Maryland. MEASUREMENTS: Data were collected by physicians at the time of each call regarding the time, day, nursing facility, reason for the call, and any orders given. Calls were also categorized as to whether documentation was necessary by defined criteria. The nursing home charts were then reviewed for the presence of documentation of 1) the issue that prompted the call, 2) physician's participation in the matter, and 3) any orders given by the physician. The relationships between the characteristics of the telephone calls and the rates of documentation were then analyzed. RESULTS: There were 248 calls from the two nursing homes during the 2-month study period. Nursing documentation of the issue that prompted the call was present for 80% of total calls. This documentation was more likely to be present with calls for change of patient status and notification of laboratory/radiograph results compared with calls for other issues (117 of 125 [94%] vs. 57 of 63 [90%] vs. 24 of 60 [40%], respectively; chi(2) = 78.3, P <0.0001). Calls that were categorized as "documentation necessary" by the Medical Director were more likely to be documented than calls that were not (132 of 150 [88%] vs. 9 of 35 [26%], respectively; chi(2) = 60.8, P <0.001). Of the 198 calls with any form of documentation, specific mention of physician participation in the communication was present in 89% of these calls. This documentation of physician participation was more likely to be present with calls for laboratory/radiograph notification than with calls for change of patient status or calls for other reasons (57 of 57 [100%] vs. 99 of 117 [85%] vs. 20 of 24 [83%], respectively; chi(2) = 10.0, P = 0.007). Physicians indicated that orders were given during 69% of calls, but orders were only documented for 79% of these interactions. No characteristics of the call were associated with likelihood of order documentation. CONCLUSIONS: In this study, documentation of issues that lead to telephone calls to physicians was not always present. Similarly, orders given by the physician were absent over 20% of the time. Nurses, physicians, and facilities should develop efficient and routine strategies to optimize rates of documentation of telephone communication with physicians. PMID- 15115580 TI - Dialysis in the old: a centenarian nursing home resident with end-stage renal disease. AB - Aging trends in the U.S. have resulted in the over 65 years age group with End Stage Renal Disease (ESRD) becoming the fastest growing segment of the population presenting for dialysis. Their profile differs from younger adults in that they present with significant comorbidity and vascular access problems, and require earlier preparation for dialysis. ESRD in the geriatric population most commonly results from diabetes, hypertension, glomerular and tubulointerstitial nephritis. Described are experiences with initiating a centenarian nursing home resident with uremia on hemodialysis; resulting in improved quality of life and illustrating that age alone should never be a barrier to receiving dialysis. Renal replacement therapy can enhance quality of life and function even in the old. A review of select literature pertinent to ESRD in the geriatric population follows. PMID- 15115581 TI - Osteoporosis for the home care physician. Part 1: etiology and current diagnostic strategies. AB - Osteoporosis affects over 20 million individuals in North America and is responsible for over 1.5 million fractures in the US. Although most cases of osteoporosis are primary, in 20% of older women and 40% of older men presenting with vertebral fractures, a secondary cause can be identified. The WHO based the diagnosis of postmenopausal osteoporosis on the presence of BMD T-score that is 2.5 standard deviations or greater below the mean for young women. The International Society of Clinical Densitometry defined male osteoporosis as BMD T score of 2.5 or greater below the mean for young men. BMD assessment at the hip and spine by DXA is the standard procedure to assess bone density. Laboratory testing in patients with low BMD is performed to exclude other conditions that could cause low BMD such as multiple myeloma, endocrinopathies and osteomalacia. Bone turnover marker levels currently do not predict bone mass or fracture risk and are only weakly associated with changes in bone mass. Subsequently, they are of limited use in the clinical evaluation of bone density changes. PMID- 15115582 TI - Quality palliative care in long-term care settings. AB - It is paramount that physicians and midlevel practitioners who care for residents in long-term care facilities be able to provide high-quality comfort care to their patients, the majority of whom are frail and suffering from chronic and progressive diseases. Physicians must be knowledgeable in the assessment, prevention, and relief of patients' physical, emotional, and spiritual distress, as well as develop appropriate attitudes, knowledge, and skills to care for patients who are in the last years of life. The provision of high-quality palliative care is the essence of long-term care medicine. PMID- 15115583 TI - Nursing home and nursing home physician: the Dutch experience. AB - Dutch nursing home care today includes a broad range of institutional and outreaching care functions. Medical care is an essential part of this care. Nursing home medicine in The Netherlands has developed as an officially acknowledged medical specialty. This is unique because The Netherlands is the only country in which nursing home medicine is a specific medical discipline. Because of this, a continuum in the medical care for the elderly has been developed: the family physician for medical care in the community, the nursing home physician for the institutionalized elderly, and the clinical geriatrician plus other medical specialists for elderly who require hospital care. This article describes the characteristics of Dutch nursing home care and nursing home medicine and the advantages of this system. The article also shows that the combination of the medical knowledge of family physician and nursing home physician can be expected to increase the quality of medical care for the disabled elderly in institutions and in the community. PMID- 15115584 TI - Continence care for frail older adults: it is time to go beyond assessing quality. PMID- 15115585 TI - End-of-life care in nursing homes: is the glass half empty or half full? PMID- 15115587 TI - Olfactory ensheathing cells: unique glial cell types? AB - Olfactory ensheathing cells (OECs) have recently been shown to have a remarkable ability to repair spinal cord injury. These cells were originally selected for transplant-mediated repair as their inherent behavior in the olfactory system is to support continual regeneration of olfactory receptor neurons throughout life. What is unique about this system is that olfactory receptor neurons, from the PNS are able to extend primary axons from the olfactory mucosa into the central nervous system (CNS) tissue of the olfactory bulb and synapse with second order neurons. This is one of the rare instances of axons crossing from the peripheral neurons system (PNS) into the CNS in the adult animal. In this paper the basic biology of these cells is described, making comparison with another promising candidate for transplant-mediated repair, the Schwann cell. The growth factor requirement for OECs is summarized detailing the influence of these factors on their antigenic and morphological characteristics. Evidence that OECs have distinct glial cell properties is provided with emphasis on their unique ability to interact with astrocytes. A brief background is given of the data obtained using OECs in transplantation studies and the resulting pros and cons discussed with emphasis on limitations of functional recovery. PMID- 15115588 TI - Repair of the injured spinal cord and the potential of embryonic stem cell transplantation. AB - Traditionally, treatment of spinal cord injury seemed frustrating and hopeless because of the remarkable morbidity and mortality, and restricted therapeutic options. Recent advances in neural injury and repair, and the progress towards development of neuroprotective and regenerative interventions are basis for increased optimism. Neural stem cells have opened a new arena of discovery for the field of regenerative science and medicine. Embryonic stem (ES) cells can give rise to all neural progenitors and they represent an important scientific tool for approaching neural repair. The growing number of dedicated regeneration centers worldwide exemplifies the changing perception towards the do-ability of spinal cord repair and this review was born from a presentation at one such leading center, the Kentucky Spinal Cord Injury Research Center. Current concepts of the pathophysiology, repair, and restoration of function in the damaged spinal cord are presented with an overlay of how neural stem cells, particularly ES cells, fit into the picture as important scientific tools and therapeutic targets. We focus on the use of genetically tagged and selectable ES cell lines for neural induction and transplantation. Unique features of ES cells, including indefinite replication, pluripotency, and genetic flexibility, provide strong tools to address questions of neural repair. Selective marker expression in transplanted ES cell derived neural cells is providing new insights into transplantation and repair not possible previously. These features of ES cells will produce a predictable and explosive growth in scientific tools that will translate into discoveries and rapid progress in neural repair. PMID- 15115589 TI - Behavioral testing after spinal cord injury: congruities, complexities, and controversies. AB - Selection and implementation of behavioral tests in spinal cord injury research is an important process, and yet few papers have focused on these issues. The critical component of any behavioral experiment is the ability to produce reliable, reproducible, and worthwhile data. Unfortunately, the difference between worthwhile and worthless data is often subtle. This paper describes factors that must be considered in order to select the most sensitive behavioral tests to match the hypothesis of the experiment and apply any test in a standardized, consistent manner. Classifications of behavioral tests, their strengths and limitations, as well as methods to overcome these limitations are discussed. Recent work in translating behavioral tests from rats to mice is also provided. The purpose of this article is to provide a framework by which behavioral testing can be standardized within and across spinal cord injury labs. PMID- 15115590 TI - Towards a definition of recovery of function. AB - In this review we consider recovery of function after spinal cord injury, and, in particular, recovery improved following intraspinal cellular transplants. Some recovery occurs spontaneously and this can be especially dramatic in neonates, supporting the notion that developing and adult spinal cord respond differently to injury. Recovery can be improved in both neonates and adults by appropriate cellular transplants into the injury site. We describe several functional tests used in animals with spinal lesions and transplants. We compare the effects of transplants of fetal tissue and genetically modified fibroblasts into neonatal and adult injury sites on recovery of motor and sensorimotor function. Fetal tissue transplants support greater recovery and elicit more regeneration in neonates than in adults. Transplants of fibroblasts modified to produce neurotrophic factors however support both recovery and axonal growth even in adults. The contribution of the transplant to recovery is shown by the loss of function that follows a second lesion just rostral to the original lesion/transplant site. The effect of the re-lesion indicates that the recovery is mediated by the presence of the transplant but the way in which transplants act to promote recovery may include a number of mechanisms, including regeneration and sprouting, neuroprotection, and modifications of organization of spared CNS structures. PMID- 15115591 TI - Stem and precursor cells in the nervous system. AB - The early-formed neural tube consists of proliferating, morphologically homogeneous cells, termed "neuroepithelial (NEP) stem cells" which generate neurons, astrocytes, and oligodenrocytes through a series of intermediate precursor cells. In addition to NEP cells, a second class of stem cells-the neurosphere-forming cell-can be isolated at later stages of development. NEP cells can differentiate into neural crest stem cells, which in turn generate PNS derivatives. NEP cells and neurosphereforming stem cells and more restricted precursors express a characteristic spectrum of markers that can be used to characterize them. Each of these cell types can be isolated from embryonic stem (ES) cell cultures, and their behavior appears similar to cells isolated at later developmental ages. The relative advantages and disadvantages of these cells for cell replacement therapy are discussed. PMID- 15115592 TI - The pathology of human spinal cord injury: defining the problems. AB - This article reviews the pathology of human spinal cord injury (SCI), focusing on potential differences between humans and experimental animals, as well as on aspects that may have mechanistic or therapeutic relevance. Importance is placed on astrocyte and microglial reactions. These cells carry out a myriad of functions and we review the evidence that supports their beneficial or detrimental effects. Likewise, vascular responses and the role of inflammation and demyelination in the mechanism of SCI are reviewed. Lastly, schwannosis is discussed, highlighting its high frequency and potential role when designing therapeutic interventions. We anticipate that a better understanding of the pathological responses in the human will be useful to investigators in their studies on the pathogenesis and therapy of SCI. PMID- 15115593 TI - Neural precursors as a cell source to repair the demyelinated spinal cord. AB - Schwann cells and neural precursor cells derived from adult human brain (subventricular zone) and from bone marrow were studied anatomically and physiologically after transplantation into the demyelinated rat spinal cord. All cell types formed myelin and restored conduction velocity. Following transection of the dorsal funiculus, Schwann cells and olfactory ensheathing cells facilitated axonal regeneration and restoration of conduction across the lesion site. There is discussion on the challenges of cell type selection and preparation for a potential clinical cell therapy study in human demyelinating diseases. PMID- 15115594 TI - A vertebral dislocation model of spinal cord injury in rats. AB - A new model of spinal cord injury (SCI) has been developed in the rat, which produces axonal and vascular injury within the spinal cord through lateral displacement of the vertebral column. An electromechanical feedback-controlled device produces the injury by displacing the vertebral column to the left hand side. The speed and lateral displacement is controllable by the user, and the resulting injury ranges from no histologically evident injury, to total disruption of the vertebral column with associated widespread axonal and vascular damage. Histological and immunohistological techniques were employed to correlate mechanical parameters with the extent of pathological injury of spinal cord. Axonal injury was most severe in the left lateral white matter, and vascular injury was concentrated in the gray matter. PMID- 15115595 TI - Electrical preconditioning attenuates progressive necrosis and cavitation following spinal cord injury. AB - This study evaluates the influence of preconditioning and subsequent electrical stimulation on the formation of primary and secondary lesions following spinal cord injury in rats. The dorsal surface of the spinal cord at the T7 level was stimulated 24 h before right-side hemisection (500 Hz, 10 pulses/train, at an inter-train interval of 10 sec for 2 h). Stimulation was again administered immediately after injury and then every 24 h for 7 days. Preconditioning electrical stimulation of the spinal cord activated reactive astrocytes, then significantly attenuated edema, progressive necrosis, and cavitation, especially in the secondary cavity lesions (24 h, 1 week, and 3 weeks post-injury). Upregulation of glial fibrillary acidic protein (GFAP) and vimentin immunoreactivity, a measure of reactive astrocytic response, were increased at 1 week after injury in the rats treated with electrical stimulation. These results suggest that preconditioning with electrical stimulation prevents the formation of secondary lesions after spinal cord injury. This beneficial effect may be related to the ability of electrical stimulation to attenuate trauma-induced cellular cascades. PMID- 15115596 TI - Traumatic injury-induced midkine expression in the adult rat spinal cord during the early stage. AB - Spinal cord injury is a debilitating condition. Midkine (MK) is involved in the generation of the central nervous system during development; however, the role of MK in the mature spinal cord has not been clarified. We examined the expression of MK, which has neurotrophic activity, before and after traumatic injury to the adult rat spinal cord. Following laminectomy, the rat spinal cord was injured at the T-9 level by applying extradural static weight-compression, in which a cylindrical compressor was used to induce complete and irreversible transverse spinal cord injury with paralysis of the lower extremities. The expression of MK was examined up to 14 days after the injury by immunohistochemical and Western blot analyses. Intense MK immunoreactivity was observed in the gray matter around the injury site but not in the necrotic lesion 1-7 days postinjury, although it was slightly positive 14 days after the injury. MK immunoreactivity was not detected in the normal spinal cord. The expression of MK was an early event, and its expression was compared to the increased production of glial fibrillary acidic protein (GFAP), a marker of reactive astrocytes, that was elevated at 2 days postinjury and continued over a 14 day period following the injury. Double immunostaining with anti-MK and anti-GFAP showed the existence of MK in the astrocytic cytoplasm. These findings suggest that MK was produced in astrocytes approximating the damaged region and may represent a reparative neurotrophic factor during the early phase of traumatic injury of the spinal cord. PMID- 15115597 TI - Spinal axonal injury transiently elevates the level of metabotropic glutamate receptor 5, but not 1, in cord-projection central neurons. AB - In investigating the effect of spinal injury on cord-projection central neurons, we found that rat rubrospinal neurons retained glutamatergic afferents and, in general, ionotropic glutamate receptor expression following spinal axotomy. Since glutamate also acts on second-messenger-coupled metabotropic receptors, the expression of group I metabotropic glutamate receptors, mGluR1 and mGluR5, was examined following similar treatment. mGluR1 expression began to decline in the perikarya 2 days postlesion and a day later in the neuropil. The decline slowed down by the fifth day and recovered in both the perikarya and neuropil 1 week postlesion. However, expression in both the perikarya and neuropil declined again and persisted up to 2 years postlesion. Similarly, the mGluR5 displayed an early transient decrease and returned to normal levels by 7 days post-lesion. However, rather than progressing to a secondary decline, the expression of mGluR5 increased to levels dramatically higher than those of control nuclei at 2-4 weeks postlesion, subsiding again by 8 weeks, and remaining low up to 2 years postinjury. Although mGluR5 has been shown to save cultured neurons from excitotoxic cell death, its elevated expression in the present model corresponds in time to an increased input/output relationship and excitability of the injured neurons as well as a period of maximal somatic shrinkage and cell loss. In addition to the cell bodies and dendrites, axon-like profiles also contain mGluR1. Their decrease following rubrospinal axotomy suggests that axonal injury may also compromise the presynaptic regulation of afferent activities onto injured cord-projection central neurons. PMID- 15115598 TI - Induction of the stress response after inflicted and non-inflicted traumatic brain injury in infants and children. AB - Rapid induction of 72-kD heat shock protein (Hsp70) is a key component of the stress response and is seen after a variety of insults to the brain including experimental hyperthermia, ischemia, seizures, and traumatic brain injury (TBI). Little is known about the endogenous stress response in pediatric patients after brain injury. Accordingly, the concentration of Hsp70 was determined in 61 cerebrospinal fluid (CSF) samples from 20 infants and children after TBI. Peak Hsp70 level were increased in TBI patients vs. controls (4.60 [1.49-78.99] vs. 2.18 [1.38-4.25] ng/mL, respectively, median (range), p = 0.01) and occurred most often on day 1 after injury. Strikingly, CSF levels of Hsp70 were positively and independently associated with inflicted vs. non-inflicted TBI (7.03 [2.30-27.22] vs. 2.06 [1.06-78.99] ng/mL, respectively, p = 0.05). Endogenous Hsp70 expression was confirmed by Western blot and immunocytochemistry using brain tissue samples removed from patients who underwent decompressive craniotomy for refractory intracranial hypertension or at autopsy. These data suggest that the endogenous stress response, as measured and quantified by the Hsp70 concentration in CSF, occurs in infants and children after TBI. The endogenous stress response is more robust in victims of child abuse, compared with patients with accidental TBI, supporting age-dependence or a difference in either injury frequency, duration, severity, or mechanism in this subgroup of TBI patients. Further studies are needed to determine the role of Hsp70 in both non-inflicted and inflicted TBI in infants and children. PMID- 15115599 TI - Continued in situ DNA fragmentation of microglia/macrophages in white matter weeks and months after traumatic brain injury. AB - Paraffin-embedded material from the pons of head-injured patients whose disability could be attributed to diffuse traumatic axonal injury, and controls, was identified from the department's archive. The cases were divided into three groups based on survival, viz Group 1 (n = 5) who survived for between 4 and 8 weeks, Group 2 (n = 5) for between 3 and 9 months, and Group 3 (n = 5) who survived for more that 12 months. Sections were stained by the TUNEL (TdT mediated UTP nick end labelling) technique, and by H&E, LFB/CV and immunohistochemically for astrocytes (GFAP) and microglia/macrophages (CD68). Microscopic abnormalities were mapped onto line diagrams of two levels of the pons and quantitation of the response determined by an eye-piece graticule placed over the medial lemmisci, cortico-spinal and transverse fiber tracts. Data were pooled by region of interest. In the H&E and LFB/CV stained sections, there was variable pallor of staining in ascending and descending fiber tracts due to loss of myelin: within these same tracts there was an astrocytosis and increased numbers of microglia/macrophages compared with controls. In the white matter tracts of the controls, there was on average 1-2 TUNEL+ cells per unit area. In contrast, there were on average 2-16 TUNEL+ cells in the cortico-spinal tracts and in the medial lemnisci of all groups of head-injured patients. CD68+ cells co located with the TUNEL+, and their number mirrored the TUNEL + staining with on average 16-30 cells per unit area in Group 1, 14-27 cells per unit area in Group 2, and 12-14 cells per unit area in Group 3. There was a statistical association between the TUNEL+ and CD68+ cells. Few changes were seen in the transverse fiber tracts of the pons. These findings indicate that most of the in situ DNA fragmentation occurred in microglia/macrophages in ascending and descending fiber tracts of the brain stem in which by conventional light microscopy there is Wallerian degeneration. However, in addition, a few TUNEL+ oligodendrocyte-like cells were also seen. PMID- 15115600 TI - Pathophysiological changes of the central auditory pathway after blunt trauma of the head. AB - It is the aim of the present paper to correlate clinical symptoms of auditory dysfunction (tinnitus, hyperacusis, hearing loss) one year on average after a blunt trauma of the head with objective audiological test results (otoacoustic emission and auditory brainstem response testing, impedance audiometry) and to compare these findings to controls without history of head trauma. Thirty-one patients (24-56 years) were included. They were largely female (n = 26). The clinical and otolaryngological examination (including otoscopy) of all patients revealed no pathological abnormalities. Pure-tone audiograms were normal with one exception (pre-existing noise-induced hearing loss) as well as tympanograms. The main auditory symptoms were tinnitus (n = 9), hyperacusis (n = 2) and a reported transient hearing loss immediately after the trauma (n = 16) (which had improved at the time of examination). The results of testing the central auditory pathway showed that the transiently evoked otoacoustic emissions (otoemissions) revealed statistically significant differences between amplitude differences of all patients as well as patients with tinnitus and controls in the linear, but not in the non-linear stimulation mode. A complete loss of stapedial reflex responses was found in 12 of the patients and a partial (irregular) loss (in at least more than two frequencies) in four additional patients. Auditory brainstem responses (ABR) were normal in all patients, but 76% had lowered loudness discomfort levels (LDL). Blunt trauma of the head can lead to auditory dyfunction, probably as a result of diffuse axonal injury of the central auditory pathway. An initial sensorineural hearing loss after the trauma (as a result of the inner ear fluid concussion) was transiently reported only. Auditory symptoms play a minor role in the so-called "postconcussive syndrome," but should be considered and evaluated fully. PMID- 15115601 TI - Potentiation of GABA(A) currents after mechanical injury of cortical neurons. AB - Numerous studies have implicated glutamate receptors, glutamate neurotoxicity, and hyperexcitation in the pathobiology of traumatic brain injury, yet much less is known about the effects of neurotrauma on inhibitory GABA channels of the brain. Using an in vitro cell injury model, we tested whether mild stretch injury altered the GABA(A) currents of cultured rat cortical neurons. The application of 1-100 microM GABA to single pyramidal neurons voltage clamped to -60 mV activated an inward current that reversed near 0 mV in solutions containing symmetrical [Cl ]. This current was inhibited by bicuculline, consistent with mediation by GABA(A) receptor channels. In injured neurons, 50 microM GABA elicited a peak current density of 41.2 +/- 2.6 pA/pF (n = 82), which was significantly larger than in uninjured control neurons, 20.2 +/- 1.7 pA/pF (n = 69, p < 0.01). The GABA(A) currents of injured neurons did not differ from those of control neurons in their sensitivity to GABA or their reversal potentials, suggesting that GABA current potentiation did not result from changes in the agonist affinity or ionic selectivity of the channels. GABA current potentiation was prevented by injuring neurons in the presence of the NMDA antagonist APV, or the CaMKII inhibitor KN93. These results thus suggest that NMDA receptor activation following neuronal injury may potentiate GABA(A) channels through the activation of CaMKII. The increase in GABA(A) receptor function observed following injury could potentially contribute to dysfunctional synaptic function and information processing as well as unconsciousness and coma following human brain trauma. PMID- 15115602 TI - Group I metabotropic receptor antagonism blocks depletion of calcium stores and reduces potentiated capacitative calcium entry in strain-injured neurons and astrocytes. AB - Antagonism of the group I metabotropic receptor subtype 1 (mGluR1) with (RS)-1 aminoindan-1,5-dicarboxylic acid (AIDA) has been shown to reduce deficits after in vivo or in vitro traumatic brain injury. We have previously demonstrated that AIDA prevents elevation of astrocyte IP3 subsequent to injury-induced activation of mGluRs and phospholipase C. Since IP3 can cause release of intracellular Ca2+ stores we tested the hypothesis that pre- or post-injury treatment with AIDA can affect (1) the depletion of Ca2+ stores which occurs soon after strain injury of cultured neurons and astrocytes and (2) the delayed potentiation of capacitative calcium entry in strain-injured neurons. Astrocyte or neuronal plus glial cultures were grown on Silastic membranes that were subjected to a 50-msec pulse of compressed gas, which caused membrane displacement and biaxial strain (stretch) injury of the adhering cells. Cells were treated 10 min before or immediately after injury with 100 microM AIDA and the intracellular free Ca2+ ([Ca2+]i) response to thapsigargin, which inhibits the ability of the stores to sequester Ca2+, was measured at 15 min or 3 h after injury. AIDA pre- or post injury treatment prevented the depletion of intracellular calcium stores at 15 min post-injury in astrocytes and neurons and reduced the potentiated neuronal capacitative calcium influx 3 h after injury. Since Ca2+ and Ca2+ stores influence many factors, including neuronal excitability, plasticity, protein synthesis, and neuronal-glial interactions, prevention of Ca2+ store depletion and subsequent exaggerated capacitative calcium entry may be an important subcellular mechanism by which antagonism of mGluR1 receptors exert an injury reducing effect. More globally, the results further emphasize the importance of altered signaling and calcium regulatory mechanisms in the immediate and delayed sequelae of traumatic brain injury. PMID- 15115603 TI - Subventricular zone neural stem cells remodel the brain following traumatic injury in adult mice. AB - Neural stem cells have recently been shown to contribute to the cellular remodeling that occurs following traumatic brain injury (TBI). Potential sources for these stem cells from within the brain include the subventricular zone of the lateral ventricles and the subgranular zone of the dentate gyrus. Using intraventricular injections of the fluorescent vital dye DiO in mice, we demonstrate that the subventricular zone population of stem cells can be reliably labeled and followed over time. By following these injections with a contralateral controlled cortical injury we demonstrate that cells from the subventricular zone migrate to the most proximally injured cortical areas. Using doublelabeling immunohistochemistry with anti-nestin, anti-GFAP, and anti-NeuN antibodies we demonstrate that labeled cells from the subventricular zone contribute primarily to the astroglial scar following injury. We do not observe any contribution to deeper areas of injury including the hippocampus. These data demonstrate that the subventricular zone contributes to brain remodeling following TBI, though neural stem cell sources outside the subventricular zone appear to play reparative roles as well. PMID- 15115604 TI - Systemic administration of 17beta-estradiol reduces apoptotic cell death and improves functional recovery following traumatic spinal cord injury in rats. AB - Recent evidence indicates that estrogen exerts neuroprotective effects in both brain injury and neurodegenerative diseases. We examined the protective effect of estrogen on functional recovery after spinal cord injury (SCI) in rats. 17beta estradiol (3, 100, or 300 microg/kg) was administered intravenously 1-2 h prior to injury (pre-treatment), and animals were then subjected to a mild, weight-drop spinal cord contusion injury. Estradiol treatment significantly improved hind limb motor function as determined by the Basso-Beattie-Bresnahan (BBB) locomotor open field behavioral rating test. Fifteen to 30 days after SCI, BBB scores were significantly higher in estradiol-treated (100 microg/kg) rats when compared to vehicle-treated rats. Morphological analysis showed that lesion sizes increased progressively in either vehicle-treated or 17beta-estradiol-treated spinal cords. However, in response to treatment with 17beta-estradiol, the lesion size was significantly reduced 18-28 days after SCI when compared to vehicle-treated controls. Terminal deoxynucleotidyl transferase-mediated UTP nickend labeling (TUNEL) staining and DNA gel electrophoresis revealed that apoptotic cell death peaked 24-48 h after injury. Also, SCI induced a marked increase in activated caspase-3 in the spinal cord, evident by 4 h after injury. However, administration of 17beta-estradiol significantly reduced the SCI-induced increase in apoptotic cell death and caspase-3 activity after SCI. Furthermore, 17beta estradiol significantly increased expression of the anti-apoptotic genes, bcl-2 and bcl-x, after SCI while expression of the pro-apoptotic genes, bad and bax, was not affected by drug treatment. Finally, intravenous administration of 17beta estradiol (100 microg/kg) immediately after injury (post-treatment) also significantly improved hind limb motor function 19-30 days after SCI compared to vehicle-treated controls. These data suggest that after SCI, 17 beta-estradiol treatment improved functional recovery in the injured rat, in part, by reducing apoptotic cell death. PMID- 15115605 TI - Traumatic axonal injury is exacerbated following repetitive closed head injury in the neonatal pig. AB - Inflicted brain injury is associated with widespread traumatic axonal injury (TAI) and subdural hematoma and is the leading cause of death in infants and children. Anesthetized 3-5-day-old piglets were subjected to either a single (n = 5) or double (n = 6, 15 min apart) rapid (<15 msec), non-impact, axial rotations of the head. Peak rotational velocities (averaging 172 rad/sec for single and 138 rad/sec for double loads) were lower than those utilized to induce severe injuries (240-260 rad/sec; Raghupathi and Margulies, 2002). At 6 h post-injury, brains were evaluated for the presence TAI using immunohistochemistry for the 200 kDa neurofilament protein (NF200). Accumulation of NF200 was observed in both contiguous (swellings) and in disconnected axons (axon bulbs) predominantly in central deep and peripheral subcortical white matter regions in the frontal, temporal, and parietal lobes of all injured piglets. Although the density of injured axons did not significantly increase after two rotational loads, the distribution of injured axons shifted from a few foci (2.2 +/- 2.3 per animal) with 1-2 swellings/bulbs following a single rotation to significantly more foci (14.7 +/- 11.9), and additional foci (2.5 +/- 1.9) containing 3 or more axon swellings/bulbs following two rotational loads. The density and distribution of injured axons following a single mild rotation were significantly reduced compared with those obtained previously following a single more severe rotational load. Collectively, these data are indicative of the graded response of the immature brain to rotational load magnitude, and importantly, the vulnerability to repeated, mild, non-impact loading conditions. PMID- 15115606 TI - A model for studies of intracranial volume pressure dynamics in traumatic brain injury. AB - The present study was undertaken to establish an experimental trauma model where it was possible to alter intracranial pressure (ICP) dynamics without raising intracranial pressure to abnormal levels and monitor metabolic disturbances with microdialysis. Thirty rats were intubated and mechanically ventilated before and after trauma. ICP was measured in the left ventricle. A weight-drop technique (21 g from 35 cm) with a brain compression of 1.5 mm was used to produce the injury. Intracranial compensatory volume was decreased 20 or 60 microL by placement of rubber film between the dura mater and bone. A bolus injection technique was used for the pressure volume response. ICP remained within normal limits for 2 h after trauma irrespective of the reduction in compensatory intracranial volume. Pressure-volume index decreased from 0.0825 +/- 0.009 to 0.0779 +/- 0.011 mL in the sham trauma and from 0.0871 +/- 0.018 to 0.0748 +/- 0.017 mL in the trauma groups (p < 0.015) when the intracranial volume was reduced by 60 microL. Intracranial compliance was not affected significantly. The present study shows that it is possible to vary ICP dynamics in a traumatic brain injury model without causing pathological increases in baseline ICP. This model may be used to study the effects of secondary insults (i.e., hypotension, hypoxia, hypercarbia, and hyperthermia) on the injured brain when ICP is normal but intracranial compensatory volume is impaired. PMID- 15115607 TI - Adenovirus vector-mediated in vivo gene transfer of brain-derived neurotrophic factor (BDNF) promotes rubrospinal axonal regeneration and functional recovery after complete transection of the adult rat spinal cord. AB - Neurotrophins have been shown to promote axonal regeneration, but the techniques available for delivering neurotrophins have limited effectiveness. The aim of this study was to evaluate the effect of adenovirus vector mediated gene transfer of brain-derived neurotrophic factor (BDNF) on axonal regeneration after spinal cord injury. We prepared adenovirus vectors encoding either beta-galactosidase (AxCALacZ) or BDNF (AxCABDNF). AxCALacZ was used to assess infection levels of the adenovirus BDNF produced by AxCABDNF was detected by Western blotting and its bioactivity was confirmed by bioassay. As a model of spinal cord injury, the rat spinal cord was completely transected at the T8 level. Immediately after transection, the vectors were injected into both stumps of the spinal cord. Axonal regeneration after transection was assessed by retrograde and anterograde tracing. In AxCALacZ-injected rats, adenovirus-infected cells were observed not only at the injected site but also in brainstem nuclei, as shown by LacZ expression. After the injection of the retrograde tracer fluorogold (FG) distal portion to the transection, AxCABDNF-injected rats showed FG-labeled neurons in the red nucleus. The anterograde tracer biotinylated dextran amine (BDA) injected into the red nucleus was also found in regenerating rubrospinal fibers distal to the transection. These tracing experiments demonstrated the regeneration of descending axons. In addition, rats of the AxCABDNF group showed significant locomotor recovery of hindlimb function, which was completely abolished by re transection. These results indicate that the recovery was caused by regeneration of rubrospinal axons, not by simple enhancement of the central pattern generator. PMID- 15115608 TI - Assessment of factors regulating axon growth between the cortex and spinal cord in organotypic co-cultures: effects of age and neurotrophic factors. AB - Axon growth failure in the central nervous system (CNS) of adult animals is thought to be attributable to several factors, including an inadequate intrinsic growth response, the presence of inhibitory molecules, and a lack of adequate neurotrophic support. Here we use a new in vitro assay system to quantitatively assess growth of axons in cortex/spinal cord organotypic co-cultures from neonatal rats. Co-cultures of cortex and spinal cord were prepared from neonatal rats at P3 or P7, and by pairing cortex and spinal cords from different ages. Axon growth from the cortex to the spinal cord was assessed using DiI tract tracing techniques. Axons could be traced from the cortex to the spinal cord in co-cultures in which both tissues were obtained from P3 animals, whereas few axons crossed the cortex/spinal cord boundary in co-cultures from P7 animals. A larger number of axons could be traced across the boundary in co-cultures from P3 animals that were treated with neurotrophins (NGF, BDNF, or NT3), whereas neurotrophins produced minimal growth enhancement in P7 co-cultures. In mixed age co-cultures of P7 cortex with P3 spinal cord, moderate numbers of axons extended between the cortex and spinal cord when cultures were treated with neurotrophins, but few if any crossing axons were detected in co-cultures of P3 cortex with P7 spinal cords. These results indicate that successful growth of axons from the cortex to the spinal cord depends on the developmental age of the tissue terrain (the spinal cord and/or the interface between cortex and spinal cord explants), and to a lesser extent on the developmental state of the cortical neurons, and that axon growth between cortex and spinal cord can be enhanced by exogenous neurotrophins. These co-cultures provide a potentially useful assay for factors that affect axon growth that is intermediate between assays based on dissociated neurons and the intact tissue terrain. PMID- 15115609 TI - Calcium entry through L-type calcium channels is essential for neurite regeneration in cultured sympathetic neurons. AB - Previous work showed that a post-neuritotomy rise in [Ca2+]i is required for regeneration. We tested the following hypotheses in cultured sympathetic neurons: (1) blocking L-type channels at the time of injury inhibits regeneration; (2) enhancing Ca2+ entry through L-type Ca2+ channels enhances regeneration; (3) L type Ca2+ channel distribution is predominantly on the soma and proximal neurites of uninjured and injured neurons. To visualize L-type Ca2+ channels and block Ca2+ influx, the fluorescent dihydropyridine antagonist, DM-BODIPY, was used. Our results show that regeneration is markedly inhibited by the antagonist when administered 20 min. prior to injury, in the presence or absence of nerve growth factor (NGF) (p < 0.0001). Severe degeneration of proximal and distal neurites was seen 48 h after injury. Regeneration was minimally inhibited by the antagonist when administered 5 min after injury (p < 0.05), but not inhibited when administered 2 or 24 h after injury (p > 0.05). We found that L-type channels are distributed ubiquitously on the soma and neurites of uninjured and injured cells, and on regenerating neurites. The addition of the L-type channel agonist, BayK8644, (1 microM) 20 min prior to injury enhanced neurite length at 24 h post-injury (p = 0.002). Blocking L-type channels did not affect the viability of uninjured or injured cells. For the first time, it has been shown that Ca2+ entry through L-type Ca2+ channels is essential for post-neuritotomy sympathetic neurite regeneration, and that this effect shows a strict temporal dependency. We also demonstrated that regeneration can be enhanced by increasing Ca2+ influx through L-type channels. PMID- 15115610 TI - Myosin binding protein C: structural abnormalities in familial hypertrophic cardiomyopathy. AB - The muscle protein myosin binding protein C (MyBPC) is a large multi-domain protein whose role in the sarcomere is complex and not yet fully understood. Mutations in MyBPC are strongly associated with the heart disease familial hypertrophic cardiomyopathy (FHC) and these experiments of nature have provided some insight into the intricate workings of this protein in the heart. While some regions of the MyBPC molecule have been assigned a function in the regulation of muscle contraction, the interaction of other regions with various parts of the myosin molecule and the sarcomeric proteins, actin and titin, remain obscure. In addition, several intra-domain interactions between adjacent MyBPC molecules have been identified. Although the basic structure of the molecule (a series of immunoglobulin and fibronectin domains) has been elucidated, the assembly of MyBPC in the sarcomere is a topic for debate. By analysing the MyBPC sequence with respect to FHC-causing mutations it is possible to identify individual residues or regions of each domain that may be important either for binding or regulation. This review looks at the current literature, in concert with alignments and the structural models of MyBPC, in an attempt to understand how FHC mutations may lead to the disease state. PMID- 15115611 TI - Peptide nucleic acid (PNA) binding-mediated gene regulation. AB - Peptide nucleic acids (PNAs) are synthetic oligonucleotides with chemically modified backbones. PNAs can bind to both DNA and RNA targets in a sequence specific manner to form PNA/DNA and PNA/RNA duplex structures. When bound to double-stranded DNA (dsDNA) targets, the PNA molecule replaces one DNA strand in the duplex by strand invasion to form a PNA/DNA/PNA [or (PNA)2/DNA] triplex structure and the displaced DNA strand exists as a single-stranded D-loop. PNA has been used in many studies as research tools for gene regulation and gene targeting. The D-loops generated from the PNA binding have also been demonstrated for its potential in initiating transcription and inducing gene expression. PNA provides a powerful tool to study the mechanism of transcription and an innovative strategy to regulate target gene expression. An understanding of the PNA-mediated gene regulation will have important clinical implications in treatment of many human diseases including genetic, cancerous, and age-related diseases. PMID- 15115612 TI - LIGHT sensitizes IFN-gamma-mediated apoptosis of HT-29 human carcinoma cells through both death receptor and mitochondria pathways. AB - LIGHT [homologous to lymphotoxins, shows inducible expression, and competes with herpes simplex virus glycoprotein D for herpes virus entry mediator (HVEM/TR2)] is a new member of TNF superfamily. The HT-29 colon cancer cell line is the most sensitive to LIGHT-induced, IFNg-mediated apoptosis among the cell lines we have examined so far. Besides downregulation of Bcl-XL, upregulation of Bak, and activation of both PARP [poly (ADP-ribose) polymerase] and DFF45 (DNA fragmentation factor), LIGHT-induced, IFNg-mediated apoptosis of HT-29 cells involves extensive caspase activation. Caspase-8 and caspase-9 activation, as shown by their cleavages appeared as early as 24 h after treatment, whereas caspase-3 and caspase-7 activation, as shown by their cleavages occurred after 72 h of LIGHT treatment. Caspase-3 inhibitor Z-DEVD-FMK (benzyloxycarbonyl-Asp-Glu Val-Asp-fluoromethylketone) and a broad range caspase inhibitor Z-VAD-FMK (benzyloxycarbonyl-Val-Ala-Asp fluoromethylketone) were able to block LIGHT induced, IFNg-mediated apoptosis of HT-29 cells. The activity of caspase-3, which is one of the major executioner caspases, was found to be inhibited by both Z DEVD-MFK and Z-VAD-FMK. These results suggest that LIGHT-induced, IFNg-mediated apoptosis of HT-29 cells is caspase-dependent, and LIGHT signaling is mediated through both death receptor and mitochondria pathways. PMID- 15115613 TI - The thymic stromal cell line MTSC4 induced thymocyte apoptosis in a non-MHC restricted manner. AB - Mouse thymic stromal cell line 4 (MTSC4) is one of the stromal cell lines established in our laboratory. While losing the characteristics of epithelial cells, they express some surface markers shared with thymic dendritic cells (TDCs). To further study the biological functions of these cells, we compared the capability of MTSC4 with TDCs in the induction of thymocyte apoptosis, using thymic reaggregation culture system. Apoptosis of thymocytes induced by MTSC4 and TDCs was measured by Annexin V and PI staining and analyzed by flow cytometry. We found that MTSC4 selectively augmented the apoptosis of CD4+8+ (DP) thymocytes. This effect was Fas/FasL independent and could not be blocked by antibodies to MHC class I and class II molecules. In addition, MTSC4 enhanced the apoptosis of DP thymocytes from different strains of mice, which implies that MTSC4-induced thymocyte apoptosis is not mediated by the TCR recognition of self peptide/MHC molecules. In contrast to MTSC4, thymocyte apoptosis induced by TDCs was MHC restricted. Thus, MHC-independent fashion of stromal-DP thymocyte interaction may be one of the ways to induce thymocyte apoptosis in thymus. Our study has also shown that the interaction of MTSC4 stromal cells and thymocytes is required for the induction of thymocyte apoptosis. PMID- 15115614 TI - LDFF, the large molecular weight DNA fragmentation factor, is responsible for the large molecular weight DNA degradation during apoptosis in Xenopus egg extracts. AB - DNA degradation is a biochemical hallmark in apoptosis. It has been demonstrated in many cell types that there are two stages of DNA fragmentation during the apoptotic execution. In the early stage, chromatin DNA is cut into large molecular weight DNA fragments, although the responsible nuclease(s) has not been recognized. In the late stage, the chromatin DNA is cleaved further into short oligonucleosomal fragments by a well-characterized nuclease in apoptosis, the caspase-activated DNase (CAD/DFF40). In this study, we demonstrate that large molecular weight DNA fragmentation also occurs in Xenopus egg extracts in apoptosis. We show that the large molecular weight DNA fragmentation factor (LDFF) is not the Xenopus CAD homolog XCAD. LDFF is activated by caspase-3. The large molecular weight DNA fragmentation activity of LDFF is Mg2+-dependent and Ca2+-independent, can occur in both acidic and neutral pH conditions and can tolerate 45 degrees C treatment. These results indicate that LDFF in Xenopus egg extracts might be a new DNase (or DNases) responsible for the large DNA fragmentation. PMID- 15115615 TI - ERK1/2 contributes negative regulation to STAT3 activity in HSS-transfected HepG2 cells. AB - Signal transducer and activator of transcription 3 (STAT3) is a recently characterized transcription factor which is essential to liver regeneration. We have previously reported that hepatic stimulator substance (HSS), a novel growth promoting substance, phosphorylated the epidermal growth factor (EGF) receptors and activated downstream Ras-MAP kinase (extracellular signal-regulated kinases, ERK1/2) cascade. However, whether HSS signal is related to STAT3 pathway remains unclear. The present study is aiming to explore the regulatory effect of activation of ERK1/2 evoked by HSS on STAT3 phosphorylation and STAT3 signaling. Human hepatoma cell line HepG2 was stably transfected with HSS cDNA and HSS expression was measured by Northern blot. The results showed that the transfection of HSS into HepG2 resulted in remarkable increase in cellular proliferation as compared with the non-transfected cells, and it was further proved that the cellular proliferation in the HSS-transfected cells was related to ERK1/2 activation. Treatment of the cells with 50 mM of PD98059, an ERK1/2 specific upstream inhibitor, resulted in ERK1/2 inactivation completely. Inhibition of ERK1/2 allowed the tyrosine of STAT3 to be phosphorylated in a dose dependent manner to PD98059. Furthermore, transient transfection of STAT3 mutant (STAT3S727A) into HSS-bearing cells could remarkably reverse the inhibitory effect of ERK1/2 on STAT3 phosphorylation. Based upon these results, it is concluded that ERK1/2 negatively modulates STAT3 phosphorylation and this function is dependent on residual serine-727 (S727) of STAT3. PMID- 15115617 TI - Inhibition of the activating signals in NK92 cells by recombinant GST-sHLA-G1a chain. AB - The soluble HLA-G1 (sHLA-G1) isoform was found to be secreted by trophoblast cells at the materno-fetal interface, which suggests that it may act as an immunomodulator during pregnancy. In this paper, we reported that GST-sHLA-G1a chain could bind to its receptor ILT-2 on NK92 cells and then the latter recruited Src homology 2 domain-containing tyrosine phosphatase-1 (SHP-1), which consequently dephosphorylated some important protein tyrosine kinases and blocked the activation of downstream molecules such as MEK and ERK so that the cytotoxicity of natural killer (NK) cells was inhibited. These results indicated that GST-sHLA-G1a chain might be exploited in new immunotherapy strategies aiming at inducing immunotolerance during allograft, xenograft and autoimmune situations. In addition, we found that modification of O-linked b-N acetylglucosamine (O-GlcNAc) was involved in NK cells' activating and inhibitory signals. This may provide a novel molecular target for inducing immunotolerance but needs further study. PMID- 15115616 TI - Binding activity of H-Ras is necessary for in vivo inhibition of ASK1 activity. AB - H-Ras is well known as one of the essential components of Ras/Raf/MEK/ERK cascade, which is a critical prosurvival signaling mechanism in most eukaryotic cells. Ras targets Raf/MEK/ERK cascade by integrating and transmitting extracellular signals from growth factor receptors to Raf, leading to the propagation of signals to modulate a serious of cellular survival events. Apoptosis signal-regulating kinase1 (ASK1) serves as a general mediator of cell death because it is responsive to a variety of death signals. In this study, we found that H-Ras interacted with ASK1 to cause the inhibition of both ASK1 activity and ASK1-induced apoptosis in vivo, which was reversed only partially by addition of RafS621A, an antagonist of Raf, whereas MEK inhibitor, PD98059, and PI3K inhibitor, LY294002, did not disturb the inhibitory effect of H-Ras on ASK-1 induced apoptosis. Furthermore, by means of immunoprecipitate and kinase assays, we demonstrated that the interaction between H-Ras and ASK1 as well as the inhibition of ASK1 activity were dependent on the binding activity of H-Ras. These results suggest that a novel mechanism may be involved in H-Ras-mediated cell survival in addition to the well established MEK/ERK and PI3K/Akt kinase dependent enhancement of cell survival. PMID- 15115618 TI - Influence of expressed TRAIL on biophysical properties of the human leukemic cell line Jurkat. AB - The cDNA fragment of human TRAIL (TNF-related apoptosis inducing ligand) was cloned into RevTet-On, a Tet-regulated and high-level gene expression system. The gene expression system was constructed in a human leukemic cell line: Jurkat. By using RevTet-On TRAIL gene expression system in Jurkat as a cell model, we studied the influence of TRAIL gene on the changes of cellular apoptosis before and after the TRAIL gene expression, which was induced by adding tetracycline derivative doxycycline (Dox). The results indicated that the cellular apoptosis ratio was largely dependent on the trail gene expression level. Moreover, it was found that the apoptosis-inducing TRAIL could cause significant changes in the biophysical properties of Jurkat cells. The cell surface charge density decreased, the membrane fluidity declined, the elastic coefficients K1 increased, and the proportion of a-helix in membrane protein secondary structure decreased. Thus, the apoptosis-inducing TRAIL gene caused significant changes on the biomechanic properties of Jurkat cells. PMID- 15115619 TI - A PSTAIRE CDK-like protein localizes in nuclei and cytoplasm of Physarum polycephalum and functions in the mitosis. AB - CDKs play key roles in controlling cell cycle progression in all eukaryotes. In plants, multiple CDKs are present, among which the best characterized CDKs are PSTAIRE CDKs. In this study, we carried out Western blot, immunoelectron microscopy and antibody treatment with an anti-PSTAIRE monoclonal antibody to explore the subcellular localization and functions of PSTAIRE CDKs in Physarum polycephalum. The results of western blot and immunoelectron microscopy showed that in P. polycephalum, a PSTAIRE CDK-like protein was 34 kD in molecular weight and located in both nuclei and cytoplasm. In nuclei, the protein was mainly associated with chromosomes and nucleoli. The expression of the PSTAIRE CDK-like protein in both the plasmodia and nuclei showed little fluctuation through the whole cell cycle. When treated with an anti-PSTAIRE monoclonal antibody at early S phase, the cells were arrested in S phase, and the mitotic onset of P. polycephalum was blocked for about 1 h when treated at early G2 phase. Our data indicated that the PSTAIRE CDK- like protein has a direct bearing on the mitosis. PMID- 15115620 TI - Anniversary of injustice: April fool's day, 1994. Will the Enactment of Bill C-6 be the birthday of equitable reproductive health care in Canada. PMID- 15115621 TI - Prenatal screening and the assessment of risk: the view from the other side. AB - Since the introduction, in 1984, of maternal serum screening for fetal aneuploidy, obstetrical practitioners and their patients have learned to cope with the challenges and limitations of risk estimation. In the instance where the "odds" are not entirely reassuring, the hazards of invasive, yet definitive, testing are weighed against the costs of uncertainty. Non-invasive prenatal screening has improved dramatically over the past 20 years, with early administration, high sensitivity, and low false positive rates as the benchmarks. With a wide array of tests at her disposal, the woman who chooses to undergo prenatal screening for aneuploidy presumably does so in the context of counselling and consent, and with the assurance that the tests offered, having evolved through the rigours of large-scale clinical trials, are as good as they can possibly be. Or does she? The following is a highly personal account of one woman's experience with prenatal screening, in particular, with the "Pandora's box" of ultrasonic soft markers for aneuploidy. The author challenges the experts in the field to ensure that all "advancements" meet the standards described above. PMID- 15115622 TI - Effects of repeated treatment failure on the quality of life of couples with infertility. AB - OBJECTIVE: To determine the impact of repeated treatment failure of clomiphene and intrauterine insemination on the quality of life of couples with infertility. METHODS: A prospective comparative study involving 3 groups of 50 couples each. The first group consisted of infertile couples with repeated treatment failure (group FT), the second group consisted of infertile couples who had never attempted any medical treatment (group NT), and the third was a control group of couples with at least 1 child and no history of infertility. Participants completed a comprehensive questionnaire to assess 16 areas of quality of life. RESULTS: The quality of life score of the control group was higher than the scores reported by groups NT and FT (P <.001). There was no significant difference in the quality of life score reported by group NT and group FT or between male and female partners. Among the women, a high quality of life was reported by 22% in group FT, 14% in group NT, and 54% in the control group. Among the men, a high quality of life was reported by 14% in group FT, 12% in group NT, and 60% in the control group. In contrast to their male partners, the women with repeated treatment failure placed higher importance on children and home than their counterparts who had not started the treatment (P <.05). CONCLUSION: Although couples with repeated treatment failure of clomiphene and intrauterine insemination do not demonstrate a lower overall quality of life than other infertile couples beginning baseline assessment, the quality of life of infertile couples is lower than that of fertile couples. PMID- 15115623 TI - Management options for preterm labour in Canada. AB - OBJECTIVES: (1) To determine the current use of treatment options for preterm labour and (2) to review the relative safety and efficacy of each class of tocolytic agent. METHODS: MEDLINE and the Cochrane Library were searched using the terms "preterm delivery," "preterm labour," and "tocolysis" alone, and in combination with the terms "betamimetics," "ritodrine," "magnesium sulfate," "calcium channel blockers," "nifedipine," "prostaglandin synthetase inhibitors," "indomethacin," "glyceryl trinitrate," "nitroglycerin," "oxytocin antagonists," and "atosiban." Randomized controlled trials (RCTs) that compared the effect of a tocolytic with a placebo or other tocolytic in women with preterm labour were selected. Trials were assessed according to Jadad's validated quality scale for assessing the quality of RCTs. Thirty two RCTs retrieved met the inclusion criteria. Information not evident from RCTs was sought from non-randomized clinical trials, cohort studies, case control studies, case series, and case reports. Data from the Canadian Survey on Tocolytic Use was obtained directly from the Society of Obstetricians and Gynaecologists of Canada (SOGC). RESULTS: The available evidence on the use of tocolytics to prolong pregnancy fails to show benefits in neonatal survival and reduced disability. This uncertainty of benefit makes the issue of fetal and maternal drug toxicity more important. There is great concern among SOGC members over the lack of effective and safe treatment options for the management of preterm labour. CONCLUSIONS: Further well-designed placebo-controlled trials are necessary to study the efficacy and safety of tocolytics in managing preterm labour. PMID- 15115624 TI - Canadian contraception consensus. AB - OBJECTIVE: To provide guidelines for health-care providers on the use of contraceptive methods to prevent pregnancy and sexually transmitted diseases. OUTCOMES: Overall efficacy of cited contraceptive methods, assessing reduction in pregnancy rate, risk of infection, safety, ease of use, and side effects; the effect of cited contraceptive methods on sexual health and general well-being; and the cost and availability of cited contraceptive methods in Canada. EVIDENCE: Medline and the Cochrane Database were searched for articles in English on subjects related to contraception, sexuality, and sexual health from January 1988 to March 2003, in order to update the Report of the Consensus Committee on Contraception published in May-July 1998. Relevant Canadian Government publications and position papers from appropriate health and family planning organizations were also reviewed. VALUES: The quality of the evidence is rated using the criteria described in the Report of the Canadian Task Force on the Periodic Health Examination. Recommendations for practice are ranked according to the method described in this Report. PMID- 15115626 TI - Epidemiology of genital herpes simplex virus infection in developed countries. AB - Comparisons of the seroepidemiology of genital herpes simplex virus (HSV) infection within and between countries are hampered by variations in tests, methods and populations sampled. Differences in seroprevalence may partly reflect variability in diagnostic efforts and healthcare awareness, expectations and utilization. To allow comparison between surveys and to improve their performance, seroepidemiological studies should use validated HSV type-specific tests, report age-specific or age-adjusted prevalence and define the period of time over which samples were collected. Despite the difficulty of comparing studies, the prevalence of HSV-2 infection varies between developed countries. Among healthy adult populations, HSV-2 seroprevalence is higher in the USA than in Europe. Furthermore, HSV-2 seroprevalence varies widely among European countries. For example, in 1989 HSV-2 seroprevalence among pregnant women was reported to be 33% in Sweden compared with 8.3% in Germany. In some, but not all, countries, HSV-2 seroprevalence appears to be increasing. In the USA, the National Health and Nutrition Examination Surveys found that HSV-2 seroprevalence increased by almost one third from 16.4% to 21.8% from 1976 to 1994 in people over 12 years old. The incidence of HSV infection is a measure of primary infection. HSV incidence is difficult to quantify, partly due to unrecognized or asymptomatic infections. However, estimates of incidence in North American and European populations range from 5 to 24 per 100 people per year. Prevention programmes should recognize that HSV-2 seroprevalence increases rapidly in early adult life. The proportion of genital herpes infections caused by HSV-1 is increasing in the developed world, possibly due to changes in oral-genital sexual behaviour and lower rates of HSV-1 acquisition in childhood. PMID- 15115627 TI - Epidemiology of herpes simplex virus type 2 infection in the developing world. AB - Herpes simplex virus type 2 (HSV-2) is a common infection in many countries, with prevalence in some regions, such as sub-Saharan Africa, higher than in the USA. Prevalence in adult general populations in sub-Saharan Africa ranges from 30% to 80% in women, and from 10% to 50% in men. Most data from Central and South America are from women, in whom HSV-2 prevalence ranges from about 20% to 40%. Prevalence in the general population in developing Asian countries appears to be lower (10-30%). In common with the developed world, HSV-2 seropositivity is uniformly higher in women than in men and increases with age. In general, HSV-2 seroprevalence is high in populations whose behaviour leads to a high risk of acquiring other sexually transmitted infections (STIs), such as STI clinic attendees and sex workers (SWs), with some African studies reporting greater than 80% HSV seropositivity in SWs. New infections are most common among young adults, a fact that should be considered when proposing and implementing measures to reduce HSV, and possibly HIV, transmission. Currently, comparison between studies is hampered by the lack of a validated type-specific serological assay that has a similar performance across a range of populations. HSV-2 is a major cause of genital ulcer disease (GUD) in the developing world. Genital herpes is a cause of morbidity and increases the risk of HIV acquisition, due to disruption of mucosal membranes. Where possible, the aetiology of GUD should be evaluated using polymerase chain reaction (PCR), while recognizing that co-pathogens can exist in a lesion. GUD management should incorporate HIV testing and antiherpetic treatment. PMID- 15115628 TI - The interaction between herpes simplex virus and human immunodeficiency virus. AB - Many studies indicate that herpes simplex virus (HSV) seropositivity increases the risk of acquiring HIV, with fewer studies also indicating that HSV-2 infection increases the risk of transmitting HIV. In a recent meta-analysis, HSV 2 infection increased the risk of HIV-acquisition two-fold. This increased risk may occur by HSV-2 reactivation disrupting the epithelial barrier and recruiting activated CD4 cells, which are target cells for HIV infection, into the lesion. In vivo and in vitro studies assessing the effect of HSV-2 on HIV transmission demonstrate that HIV-infected CD4 cells are recruited to HSV-infected lesions and that HSV regulatory proteins (ICP0, ICP4, VP16) may upregulate HIV replication, thus increasing the frequency and titre of mucosal HIV shedding. This may occur during both clinical and asymptomatic HSV reactivation. Plausibly, antiherpetic therapy could reduce HIV transmission by decreasing HIV plasma load and/or mucosal HIV shedding, but a proof-of-concept trial is needed to demonstrate this. It also appears that individuals co-infected with HIV and HSV-2 have more frequent HSV recurrences than individuals infected with HSV-2 alone. There is a strong correlation between decreasing CD4 count and increasing rates of HSV reactivation, suggesting that reactivation is linked to immunosuppression. The IHMF recommends that individuals with HIV should be serologically tested for HSV 2. HSV-2 infection should be targeted as a modifiable risk factor for HIV acquisition by testing, counselling and preventing acquisition through behavioural interventions, treatment and antiviral suppression. PMID- 15115630 TI - The theoretical basis of stigma as applied to genital herpes. AB - This paper defines stigma and its characteristics, outlines strategies and consequences of stigma management, describes the theoretical basis of stigma, and offers methodological considerations for those applying stigma theory to the domain of genital herpes. Stigma is an interactional process, defined within societies, in which particular social identities are collectively devalued. The subjective experience of stigma may vary due to features associated with the stigmatizing condition (e.g. concealability, course, strain, aesthetic qualities, cause and peril). The interpersonal management of stigma may include secrecy, withdrawal, covering, informing or disclosing. Future research addressing herpes associated stigma should benefit from theoretical frameworks including attribution theory, social-cognitive theory, preoccupation model of secrecy and the illness intrusiveness framework, although the difficulty in identifying and recruiting stigmatized individuals and the lack of domain-specific measures of herpes-related stigma remain barriers to progress. PMID- 15115631 TI - The effects of stigma on genital herpes care-seeking behaviours. AB - Stigma affects both herpes-infected and uninfected people. The stigma associated with genital herpes may influence a person's decision to seek care or disclose the infection to others. For uninfected people, the threat of the stigma associated with sexually transmitted infections (STIs) is an integral tool of clinical and public health approaches to genital herpes prevention. This paper reviews concepts of stigma in relation to the prevention and treatment of genital herpes and other STIs, and calls for those involved in STI care to weigh the extent that public health response is altruistic, moral and effective against the substantial, although incomplete, evidence of harm caused by stigma. PMID- 15115632 TI - Vaccination strategies to prevent genital herpes and neonatal herpes simplex virus (HSV) disease. AB - Vaccination strategies to prevent genital and neonatal herpes simplex virus (HSV) disease have a history of apparent efficacy in animal studies followed by failure in clinical trials. Further study of the immune response induced by natural HSV infections in both adults and neonates will provide insight into the requirements for vaccination against acute disease and recurrences. Lessons can also be learnt from the recent partial success of a HSV-2 glycoprotein D vaccine coupled with monophosphoryl lipid A adjuvant, which induced protection from clinical disease in HSV-1 and HSV-2 seronegative women. Its efficacy has been attributed to enhancement of Th-1 immunity by the adjuvant. Newer vaccine vectors including DNA vaccines, recombinant viral vaccines and specific HSV mutants are being developed but better animal models are required for more rapid progress. This review examines the history of HSV vaccine development, describes recent progress towards an effective prophylactic vaccine against HSV and outlines further improvements required to make current vaccines immunogenic to a wide population. PMID- 15115634 TI - Anti-inflammatory agents for the treatment of musculoskeletal pain and arthritis. AB - Pain produced by musculoskeletal disorders commonly misconceived as having mechanical etiology often is caused by inflammatory mechanisms. Simple analgesics (ie, those that lack anti-inflammatory action) often are used to treat musculoskeletal pain when an anti-inflammatory analgesic may be more effective for the painful condition. This review addresses the anti-inflammatory agents available for the symptomatic management of common inflammatory musculoskeletal conditions including osteoarthritis, rheumatoid arthritis, and low back pain. PMID- 15115635 TI - Prophylactic migraine therapy: emerging treatment options. AB - In this paper, new treatment options for migraine prevention are reviewed. An overview about migraine pathophysiology is provided and current indications for migraine prevention and new and upcoming preventive medications are discussed briefly. Data are presented on topiramate, levetiracetam, zonisamide, botulinim toxin, tizanidine, nefazodone, lisinopril, candesartan, carabersat, petasites, and coenzyme Q. PMID- 15115636 TI - Strategies for finding new pharmacological targets for neuropathic pain. AB - Chronic pain of neuropathic origin is an unmet therapeutic challenge that may require the development of novel drug entities. The resources involved in developing novel drugs and testing them in clinical trials are so large that few candidates can be fully tested in practice. A poor choice can have major repercussions. At the same time, the research community has identified large numbers of potential pharmacological targets for analgesia in recent years, with individual investigators promoting favorites with increasing zeal. Because of the uncertain predictive value of animal models of neuropathic pain, it is essential to consider rational strategic approaches to selecting the most likely candidate targets. PMID- 15115637 TI - New and emerging pharmacological targets for neuropathic pain. AB - Increasing knowledge of the molecular consequences of nerve injury and the availability of genome databases has greatly increased the range of potential targets for the pharmacological management of neuropathic pain. Controlling neuronal sensitization and the associated alterations in gene expression, protein modification, and neuronal excitability is the key to managing neuropathic pain. Control of neuronal sensitization can occur through inhibition of nerve injury associated production of cytokines, activation of glial cells, modulation of potassium channel subtypes, mitogen-activated protein kinases, the ubiquitin proteasome system, or the protection and amplification of spinal cord dorsal horn inhibitory systems. These new and already established targets promise unparalleled opportunities for the prevention, management, and resolution of persistent pain states following nerve injury. PMID- 15115638 TI - Targeted peripheral analgesics therapy for neuropathic pain. AB - The term targeted peripheral analgesics has been suggested to describe analgesics with a mechanism of action that appears to be primarily through reducing pain transmission within the peripheral nervous system. Key differences between targeted peripheral (topical) and systemic analgesics and the difference between topical and transdermal analgesics are discussed in this article. A review of the clinical conditions, which have been reported to respond to targeted peripheral analgesics, also is described in detail. PMID- 15115639 TI - Recent advances in clinical use of opioids. AB - Opioid therapy for pain is the subject of numerous randomized clinical trials. Opioids are being developed for delivery by a wide variety of mechanisms. New opioids are becoming available for clinical use. This review surveys recent developments in these clinical trials and provides an overview of what may be expected in the near future for opioid management of pain. PMID- 15115640 TI - Neuromodulating drugs for the symptomatic treatment of neuropathic pain. AB - Significant improvement of neuropathic pain has been achieved with studies that have demonstrated efficacy of newer anticonvulsants in relieving this type of pain, by having a neuromodulatory effect on the hyperexcitable damaged nervous system. Two drugs from this class, gabapentin and lamotrigine, have been submitted to a number of clinical trials. Ease of use and broad therapeutic range, in addition to demonstrated efficacy, make gabapentin the drug of choice for most neuropathic pain disorders. Lamotrigine is well tolerated when it is titrated slowly, which also is the way to avoid the development of a rash. Pregabalin, the newest agent that has demonstrated efficacy in the treatment of post-herpetic neuralgia, is awaiting approval. A number of available anticonvulsants are undergoing clinical trials and many drugs with neuromodulatory properties are being considered for further development. PMID- 15115641 TI - Pharmacokinetic studies in migraine: what questions should physicians ask? AB - Migraine is a benign self-limiting condition for which the objective of treatment is to maximize the patient's quality of life by reducing the frequency of attacks and attenuating the pain and suffering of an individual attack as quickly and safely as possible. Evaluating the efficacy of antimigraine treatments, particularly those for the management of the acute attack, is relatively easy because acute symptomatic benefit is the criterion of success. For prophylaxis, therapeutic success generally is measured by relatively simple parameters such as attack frequency. Given that the clinical response to antimigraine therapies relatively is readily assessed by history, the role of more sophisticated investigations such as pharmacokinetic studies may not be immediately clear. This paper aims to provide clinicians with a guide to interpretation of such studies. PMID- 15115642 TI - Sporadic hemiplegic migraine. AB - Hemiplegic migraine may be familial or sporadic. Both forms share a similar spectrum of clinical presentations and genetic heterogeneity. Current data support the concept that sporadic and familial cases should be thought of as similar, but separate disorders. Sporadic cases are more difficult to diagnose and often require several investigations to rule out more ominous possibilities. Therapeutic options are limited, but future genetic research may elucidate pathophysiologic mechanisms that may, in turn, yield more specific treatments. PMID- 15115643 TI - Menstrual migraine: a review of prophylactic therapies. AB - Menstrual migraine is commonly encountered in women who are experiencing attacks of migraine without aura. It remains controversial whether attacks of menstrually associated migraine are more severe and have a longer duration than non menstrually associated attacks. The pathogenesis of menstrual migraine is not understood completely, but it may be related to estrogen withdrawal or prostaglandin release. Preventative therapies may be considered in those who have failed abortive medications or have attacks lasting longer than 2 days. They can be administered short-term during the perimenstrual time period or continuously throughout the menstrual cycle. Short-term prophylactics should be tried first because menstrual migraines generally last for 1 to 4 days only. Continuous prophylactics may be considered in those with attacks refractory to short-term therapies. PMID- 15115644 TI - Toward a molecular genetic classification of familial hemiplegic migraine. AB - The genetics of migraine is a fascinating and rapidly moving research area. Familial hemiplegic migraine, a rare subtype of migraine with a Mendelian pattern of inheritance, is caused by mutations in the chromosome 19 CACNA1A gene or in the chromosome 1 ATP1A2 gene. Familial migraine variants are classified on the basis of clinical, descriptive criteria, but this is insufficient. In the future, a diagnostic classification based on mutation-analysis is needed. PMID- 15115645 TI - Anticonvulsants in migraine. AB - Migraine is a common, disabling disorder that often requires preventive treatment. The decision to treat migraine preventively generally is based on disability, problems with acute medicines, patient preference, risk of acute medication overuse, special circumstances, and concern that high migraine attack frequency may be a risk factor for chronic daily headache. Migraine and epilepsy are comorbid episodic central nervous system disorders that can have stereotyped symptoms with negative and positive phenomena. Controlled trials have demonstrated the efficacy of anticonvulsants in migraine prevention. Valproic acid, topiramate and, to a lesser extent, gabapentin, have demonstrated efficacy in randomized, placebo-controlled trials. Lamotrigine may be effective at controlling migraine aura, but has not demonstrated effectiveness at controlling migraine headache. Anticonvulsants are a useful option for the preventive treatment of migraine. PMID- 15115646 TI - Management of in-transit melanoma of the extremity with isolated limb perfusion. AB - In-transit metastases for melanoma are a type of stage III regional metastatic disease that are intradermal or subcutaneous nodules growing within lymphatics and not in nodal basins. If the initial diagnosis is a limited number of in transit metastases (1-3 nodules), the optimal management is simple surgical excision with minimal negative margins and primary closures and appropriate staging to look for any distant metastases. There is no role for wide excision of in-transit lesions as there is for primary melanoma because the entire extremity or that region of the body is at risk for recurrence. Patients who are diagnosed with additional lesions in a short period of time or patients who at initial diagnosis have large numbers of nodules are candidates for isolated limb perfusion (ILP). ILP is a regional administration of high-dose chemotherapeutics within an extremity using a cardiopulmonary bypass machine similar to cardiac surgery. Once isolation is obtained surgically, the limb is heated to what is considered mild hyperthermia (38.5 degrees -40 degrees C), then chemotherapeutics are administered at very high concentrations for a 60- to 90-minute treatment. The drug recirculates and, at the end of the treatment period, it is flushed from the extremity and the circulation is re-established. The optimal regimen is melphalan dosed per limb volume (10 mg/L limb volume for lower extremities and 13 mg/L limb volume for upper extremities) with mild hyperthermia for 60 minutes. Using this regimen, overall response rates between 80% and 90% and complete response rates between 55% and 65% can be obtained. The duration of response is typically 9 to 12 months and a subgroup of complete responders, which is 20% to 25% of the total patient population, typically have sustained complete responses. The major toxicities are skin erythema, myopathy, and peripheral neuropathy. There have been several studies adding high-dose tumor necrosis factor to ILP, but there is no clear benefit in the treatment of melanoma. Other new approaches include isolated limb infusion as a percutaneous procedure to avoid the surgical toxicity. PMID- 15115647 TI - Sentinel lymph node mapping and selective lymphadenectomy: the standard of care for melanoma. AB - Selective sentinel lymphadenectomy (SSL) should be considered a standard of care approach for staging patients with primary invasive melanoma 1 mm or greater. It is essential that multidisciplinary teams should master the techniques of preoperative lymphoscintigraphy, intraoperative lymphatic mapping, and pathologic evaluation of the sentinel lymph nodes (SLNs). An SLN may be blue, hot, or any lymph node (LN) greater than 10% of the in-vivo count of the hottest LN. An enlarged or indurated LN should be removed because it may contain metastatic cancer cells that block blue dye or radiotracer entry. Frozen sections are not recommended. Surgeons who use isosulfan blue dye should be cognizant of treatment for a potentially fatal reaction. Prophylactic LN dissection should not be performed if a SSL can be performed as a staging procedure. A complete LN dissection is performed if the SLN is positive. It is important to follow the clinical outcome of patients undergoing SSL, thus its role can be further defined. PMID- 15115648 TI - Merkel cell carcinoma. AB - Merkel cell carcinoma is a rare cutaneous neoplasm most commonly affecting the head and neck of elderly white patients. Even with treatment, Merkel cell carcinoma has a strong propensity toward local recurrence, lymphatic spread, and distant metastasis. Because of its rarity and the subsequent lack of well controlled clinical trials, no single standard of care exists for the treatment of this aggressive tumor. In our institution, primary lesions are excised with wide margins or by Mohs' micrographic surgery. After local removal, the excision site is treated locally with external radiation therapy. Sentinel lymph node mapping and biopsy are performed. Patients with tumor within a sentinel lymph node undergo lymph node dissection and radiation to the lymphatic basin. Adjuvant chemotherapy is offered to high-risk patients with local disease and to patients with metastases. Patients with distant metastases are treated with a combination of salvage chemotherapy and radiation therapy. PMID- 15115649 TI - Therapy for mycosis fungoides. AB - Treatment of mycosis fungoides (MF) is indicated to reduce symptoms, improve clinical appearance, prevent secondary complications, and prevent progression of disease, all of which may have an impact on survival. Treatment of MF includes topical and systemic therapies, which can be administered alone or in combination. Psoralen and ultraviolet A radiation is effective in early-stage MF, inducing complete remissions in most patients. Psoralen and ultraviolet A radiation may also be combined with low doses of interferon (IFN)-alpha to treat stage I/II disease. However, early aggressive therapy with radiation and chemotherapy does not improve the prognosis. Local radiotherapy or total skin electron beam irradiation has been used with success to control advanced skin disease. Extracorporeal photopheresis may also be used successfully, but it is not generally available. Once the disease becomes refractory to topical therapy, IFN-alpha single-agent or combination chemotherapy may be administered, but the duration of response is often less than 1 year and ultimately all patients will relapse and become refractory. Among chemotherapeutic agents, pentostatin, gemcitabine, and liposomal doxorubicin seem to be particularly effective. Response rates after combined modality therapy with total skin electron beam irradiation and chemotherapy/IFN-alpha appear similar to response rates of chemotherapy alone. Therefore, there is a great need for the further development of novel emerging treatment modalities, such as retinoids (ie, bexarotene) and immunotherapeutic agents (ie, cytokines, tumor vaccines, and monoclonal antibodies), all of which appear to have significant therapeutic potential in patients with MF. Biologically based therapies may reduce the need for genotoxic therapies, such as cytostatics and radiotherapy. PMID- 15115650 TI - Second neoplasms in patients with chronic lymphocytic leukemia. AB - Second malignancies occur with increased frequency in patients with chronic lymphocytic leukemia (CLL) regardless of treatment, but they may be more frequent and more aggressive after nucleoside analog therapy of CLL. In as many as 33% of patients with CLL who develop a second malignancy, a spontaneous remission of CLL precedes the diagnosis of the second malignancy by months or years. Richter's syndrome, whether manifested by anaplastic large cell lymphoma or Hodgkin's disease, is not truly a second malignancy because the CLL clone appears to be involved. However, all other malignancies developing in patients with CLL appear to be derived from a different clone. CLL and the second malignancy may have a common viral etiology. Second malignancies in patients with CLL should be treated as they would be in patients without CLL. PMID- 15115651 TI - An update of novel therapeutic approaches for multiple myeloma. AB - Multiple myeloma (MM) remains an incurable malignancy, despite conventional and high-dose therapies, and novel biologically based treatment approaches are urgently needed. Recent studies have characterized the molecular mechanisms by which MM cell/host bone marrow (BM) interactions regulate tumor cell growth, survival, and migration in the BM milieu. These studies have not only enhanced our understanding of disease pathogenesis, but they have also provided the framework for a new treatment paradigm targeting the MM cell in its BM microenvironment to overcome drug resistance and improve patient outcome. Clinical trials are confirming the remarkable activity and improved tolerability of some of the new agents identified through this paradigm, providing exciting evidence of translational success in MM. PMID- 15115652 TI - Waldenstrom macroglobulinemia. AB - Waldenstrom macroglobulinemia (WM) is a low-grade lymphoproliferative disorder characterized by the presence of an immunoglobulin M monoclonal protein in the blood and monoclonal small lymphocytes and lymphoplasmacytoid cells in the marrow. The disease is uncommon and there is a lack of clear diagnostic criteria. WM is treatable but not curable and long-term survival is possible. Therefore, the treating physician needs to carefully balance the risks and benefits of treatment. Treatments are aimed at relieving symptoms resulting from marrow infiltration and the hyperviscosity syndrome. Therapies available for initiation of treatment include alkylating agents, purine nucleoside analogs, and rituximab. Chlorambucil has been the mainstay of treatment for many years and remains useful, especially in older patients. Rituximab has become an important new therapy for this disease because of its positive treatment responses, acceptable toxicity, and lack of therapy-associated myelosuppression and myelodysplasia. Currently, rituximab is being combined with chemotherapy. Other options of treatment include interferon and corticosteroids. Emerging therapies include stem cell transplantation (autologous and allogeneic) for younger patients. Currently, there are few comparative data on which to state an absolute opinion concerning the best available treatment for patients with WM. PMID- 15115653 TI - Treatment of POEMS syndrome. AB - POEMS (polyneuropathy, organomegaly, endocrinopathy, M protein, and skin changes) syndrome is a rare multisystemic paraneoplastic syndrome driven by an underlying plasma cell dyscrasia. More than 95% of patients will have monoclonal lambda sclerotic plasmacytoma(s) or bone marrow infiltration. Recognition of the complex of a combination of peripheral neuropathy, organomegaly, endocrinopathy, monoclonal plasmaproliferative disorder, skin changes, sclerotic bone lesions, Castleman disease, thrombocytosis, papilledema, peripheral edema, pleural effusions, ascites, fingernail clubbing, and white nails is the first step in effectively managing the disease. Once a patient has been completely evaluated, each component of the disease should be addressed, while finalizing a treatment plan for the underlying plasma cell proliferative disorder. In patients with a dominant sclerotic plasmacytoma, first-line therapy should include radiation to the lesion. Retrospective analysis and personal experience would dictate that systemic therapy be considered for patients with diffuse sclerotic lesions or absence of any bone lesion and for patients who have not demonstrated stabilization of their disease 3 to 6 months after completing radiation therapy. Treatments with demonstrated benefit include corticosteroids, low-dose alkylator therapy, and high-dose chemotherapy with peripheral blood stem cell transplant. Until the pathogenesis is fully understood, these are the mainstays of treatment for patients with POEMS syndrome. PMID- 15115655 TI - The neuronal primary cilium--an extrasynaptic signaling device. AB - Many, but likely most, neurons in the central nervous system have a nonmotile "primary" cilium extending like an antenna or finger from one of the pair of centrioles in the cell's centrosome into the extracellular space. Since their discovery over 100 years ago, these organelles have been either dismissed as functionless relicts of a bygone era or more often simply ignored. However, it has long been known that the photoreceptor-bearing outer segments of retinal rods and cones are modified primary cilia and it has recently been found that kidney cells' primary cilia are sensitive flowmeters the disabling of which causes polycystic kidney disease. It has also been recently shown that somatostatin sst3 receptors and serotonin 5-HT(6) receptors are selectively sited on neurons in various parts of the rat brain. It seems likely that these selectively receptored neuronal primary cilia will turn out to be the forerunners of a family of cell signaling devices that help drive various brain functions by sending signals into their own cells and into adjacent cells through gap junctions and via conventional chemical synapses. PMID- 15115656 TI - Structure and regulation of MAPK phosphatases. AB - MAP kinases (MAPKs), which control mitogenic signal transduction in all eukaryotic organisms, are inactivated by dual specificity MAPK phosphatases (DS MKPs). Recent studies reveal that substrate specificity and enzymatic activity of MKPs are tightly controlled not only by the conserved C-terminal phosphatase domain but also by an N-terminal (NT) kinase-binding domain. Notably, MKPs that consist of a kinase-binding domain and a phosphatase domain exhibit little phosphatase activity in the absence of their physiological substrates. MKP binding to a specific MAPK results in enzymatic activation of the phosphatase in a substrate-induced activation mechanism. This direct coupling of inactivation of an MAPK to activation of an MKP provides a tightly controlled regulation that enables these two key enzymes to keep each other in check, thus guaranteeing the fidelity of signal transduction. This review discusses the recent understanding of structure and regulation of the large family of dual specificity MKPs, which can be divided into four subgroups according to their functional domains and mechanism of substrate recognition and enzymatic regulation. Moreover, detailed comparison of the structural basis between this unique substrate-induced activation mechanism and the common auto-inhibition mechanism is provided. PMID- 15115657 TI - Epstein-Barr virus latent membrane protein 1 modulates epidermal growth factor receptor promoter activity in a nuclear factor kappa B-dependent manner. AB - The Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) oncoprotein may cause multiple cellular changes including the induction of epidermal growth factor receptor (EGFR) expression and activation of the NFkappaB transcription factor. LMP1 increases the levels of both EGFR protein and mRNA, but does not stabilize EGFR mRNA. Thus, the effects of LMP1 are likely to be mediated by the direct activation of the EGFR promoter. In this study, induction of LMP1 increased the EGFR in both protein and promoter levels in a dose-dependent manner using tetracycline-regulated LMP1 expression in nasopharyngeal carcinoma (NPC) cell line. Mutational analysis of the LMP1 protein indicated that the C-terminal activation region-1 (CTAR1) domain was mainly involved in the EGFR promoter induction, while CTAR2 was necessary but not sufficient to induce EGFR promoter. Inhibition of LMP1-mediated NFkappaB activation by constitutive repressive IkappaBalpha marginally decreased EGFR promoter activity using transiently transfected IkappaBalpha dominant negative mutant. Promoter mutagenesis analysis demonstrated that two putative NFkappaB binding sites of EGFR promoter were very necessary for the transcriptional activity of EGFR induced by LMP1, the proximal NFkappaB binding site was more important than the distal NFkappaB binding site, and both NFkappaB binding sites played a cooperative role. Taken together, Epstein-Barr virus latent membrane protein 1 modulated the EGFR promoter activity in a NFkappaB-dependent manner. PMID- 15115658 TI - NHERF2 increases platelet-derived growth factor-induced proliferation through PI 3-kinase/Akt-, ERK-, and Src family kinase-dependent pathway. AB - Platelet-derived growth factor (PDGF) has multiple functions including inhibition of apoptosis and promotion of cell proliferation. In this study, we show that Na(+)/H(+) exchanger regulatory factor 2 (NHERF2) binds to the carboxyl-terminal PDZ domain-binding motif of the PDGF receptor through a PDZ domain-mediated interaction, and evaluate the consequence on PDGF-induced proliferation. Stable transfection with NHERF2 increased the PDGF-induced phosphorylation of ERK and Akt in Rat1 embryonic fibroblasts. The phosphorylation of Akt was blocked by pretreatment with LY294002, a PI-3-kinase inhibitor, in both Rat1/NHERF2 and Rat1/vector cells. In Rat1/vector cells, PDGF-induced phosphorylation of ERK was completely inhibited by pretreatment with PD98059, a MEK inhibitor. In contrast, the NHERF2-dependent increase of ERK phosphorylation was not affected by pretreatment with PD98059 in Rat1/NHERF2 cells. Thus, the NHERF2-dependent increase of ERK phosphorylation occurs in a MEK-independent fashion. Pretreatment with PP2, a specific inhibitor of Src family tyrosine kinase, completely blocked the NHERF2-dependent increase of the phosphorylation of ERK and Akt, suggesting that NHERF2 up-regulates Erk phosphorylation through a Src family kinase dependent pathway. Consistent with these results, the PDGF-induced thymidine incorporation was increased in Rat1/NHERF2 cells, and the NHERF2-dependent increase of thymidine incorporation was prevented by treatment with LY294002 and PP2 but not with PD98059. These results suggest that NHERF2 stimulates PDGF induced proliferation by increasing PI-3-kinase/Akt, MEKindependent ERK, and Src family kinase-mediated signaling pathways. PMID- 15115659 TI - Activation of extracellular signal-regulated kinase couples platelet-activating factor-induced adhesion and delayed apoptosis of human neutrophils. AB - Platelet-activating factor (PAF) promotes adhesion of neutrophil granulocytes to the endothelium, which is also linked to neutrophil survival. Here we report that PAF can prolong neutrophil survival by suppressing spontaneous apoptosis. PAF induced concurrent activation of the Ras/Raf-1/mitogen-activated protein kinase kinase (MAPKK)/extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinase/Akt pathways. ERK activation tightly correlated with up-regulation of CD11b/CD18 expression and beta(2)-integrin-dependent homotypic adhesion. These actions of PAF were markedly attenuated by the MAPKK/ERK inhibitor PD98059, but not by the phosphatidylinositol 3-kinase inhibitor wortmannin. By contrast, concurrent activation of ERK and Akt was required to inhibit caspase-3 activation and consequently to delay apoptosis. Consistently, pharmacological inhibition of either ERK or Akt partially reversed the anti-apoptotic action of PAF; however, they did not produce additive inhibition. These results indicate that PAF-induced activation of ERK contributes to both the expression of the pro-adhesive phenotype and repression of neutrophil apoptosis, thereby amplifying the inflammatory response. PMID- 15115660 TI - Synergistic effect of endothelin-1 and cyclic AMP on glucose transport in 3T3-L1 adipocytes. AB - We have demonstrated previously that chronic exposure to endothlin-1 enhances glucose transport in 3T3-L1 adipocytes via augmented GLUT1 mRNA and protein accumulation. In the present study, we further examined the combined effect of endothelin-1 (ET-1) and cAMP on glucose transport. In cells pretreated with ET-1 and 8-bromo cAMP for 8 h, a synergy between these two agents on glucose uptake was found. Insulin-stimulated glucose transport, on the other hand, was only slightly affected. The synergistic effect of these two agents was suppressed in the presence of cycloheximide and actinomycin D. Immunoblot and Northern blot analyses revealed that GLUT1 protein and mRNA levels were both increased in cells pretreated with both ET-1 and 8-bromo cAMP, greater than the additive effect of each agent alone. Further examination demonstrated that the stability of GLUT1 mRNA was markedly enhanced in the presence of both ET-1 and cAMP. To investigate the transcriptional activation of Glut1 gene, transient transfection of cells with luciferase reporter construct driven by Glut1 promoter was performed. We found that Glut1 transcription was also increased by ET-1 and cAMP in a synergistic fashion. In addition, similar synergy between ET-1 and beta adrenergic agonists on glucose transport was found. The synergistic action of ET 1 with 8-bromo cAMP to enhance glucose transport was inhibited by GF109203X, a selective protein kinase C (PKC) inhibitor, and was mimicked by 4beta-phorbol 12beta-myristate 13alpha-acetate (PMA), a PKC activator. Furthermore, PMA was found to act synergistically with 8-bromo cAMP to induce Glut1 transcription and ET-1 was shown to activate novel PKCdelta and PKC. Taken together, these results indicate that ET-1 may act with cAMP in a synergistic way to increase glucose transport, probably through enhanced GLUT1 expression via a PKC-dependent mechanism. PMID- 15115661 TI - Gbetagamma signaling and Ca2+ mobilization co-operate synergistically in a Sos and Rac-dependent manner in the activation of JNK by Gq-coupled receptors. AB - The mechanism by which G(q)-coupled receptors stimulate the c-Jun N-terminal kinase (JNK) activity has not been fully delineated. Here, we showed that stimulation of endogenous G(q)-coupled receptors in human hepatocarcinoma HepG2 cells resulted in an Src family kinase- and Ca(2+)-dependent JNK activation. Cos 7 cells transfected with HA-tagged JNK and various G(q)-coupled receptors also exhibited similar characteristics and provided further evidence for the involvement of Gbetagamma, an upstream intermediate for Src family kinases. The Ca(2+) and Gbetagamma signals operate in a high degree of independence. Transient expression of Gbetagamma subunits and elevation of cytoplasmic Ca(2+) level by thapsigargin activated JNK in a synergistic fashion. JNK activities triggered by G(q)-coupled receptors, Gbetagamma and thapsigargin were all suppressed by dominant negative (DN) mutants of Son of sevenless (Sos) and Rac. We propose that the co-operative effect between Gbetagamma-mediated signaling and the increased intracellular Ca(2+) level represents a robust mechanism for the stimulation of JNK by G(q)-coupled receptors. PMID- 15115662 TI - JNK signaling involved in the effects of cyclic AMP on IL-1beta plus IFNgamma induced inducible nitric oxide synthase expression in hepatocytes. AB - cAMP significantly inhibits IL-1beta+IFNgamma-induced iNOS gene expression in hepatocytes, but the signaling pathways responsible for the effect are not known. PKA inhibitors, H89, PKI, and KT5720, had no effect on the recovery of the inhibitory effects of cAMP on cytokine-induced hepatocyte iNOS expression and activity. The JNK inhibitor, SP 600125, effectively reversed the inhibitory effects of cAMP on iNOS expression and significantly increased iNOS promoter activity. A cAMP analogue, dbcAMP, significantly induced JNK signaling and increased AP-1 binding activity in hepatocytes. The JNK activator, anisomycin, inhibited iNOS expression and transcription in hepatocytes as well as AP-1 binding activity; and SP600125 reversed this effect of anisomycin. Overexpression of c-Jun in hepatocytes inhibited IL-1beta+IFNgamma-induced nitrite accumulation and iNOS promoter activity while dominant negative c-Jun partially reversed the inhibitory effects of cAMP on nitrite accumulation. We conclude that JNK signaling plays an important role in the inhibitory effects of cAMP on IL 1beta+IFNgamma-induced iNOS gene expression in cultured hepatocytes. PMID- 15115663 TI - The Shp-1 and Shp-2, tyrosine phosphatases, are recruited on cell membrane in two distinct molecular complexes including Ret oncogenes. AB - The Shp-2 and Shp-1 non-transmembrane tyrosine phosphatases display different and even opposing effects on downstream signaling events initiated by Ret activation. By using rat pheochromocytoma-derived PC12 cells, here we studied the interactions of Shp-2 and Shp-1 with two activated mutants of Ret receptor, Ret(C634Y) and Ret(M918T). Each of these mutated receptors causes inheritance of distinct cancer syndromes, multiple endocrine neoplasia (MEN) type 2A and type 2B, respectively. We show that: (i) both Shp-1 and Shp-2 are associated to a multiprotein complex that includes Ret mutants; (ii) the Shp-1-Ret complexes are distinct from Shp-2-Ret complexes, and these complexes are differently distributed inside and outside lipid rafts; (iii) constitutively activated Ret proteins neither directly bind to nor are substrates of these phosphatases. Our results well support the evidence that Ret complexes within and outside rafts mediate distinct biological functions, and indicate that the presence of either Shps participates to determine such functions. PMID- 15115664 TI - Aspirin resistance and diabetic angiopathy: back to the future. PMID- 15115665 TI - Reduced sensitivity of platelets from type 2 diabetic patients to acetylsalicylic acid (aspirin)-its relation to metabolic control. AB - Aspirin (acetylsalicylic acid, ASA), which is recommended for primary and secondary prevention in diabetes mellitus (DM), has been shown to have a lower antiplatelet activity in diabetic patients. We conducted a crossover designed observational study to evaluate whether there is an association between the parameters relevant to metabolic control of diabetes and platelet sensitivity to aspirin in type 2 diabetic patients. Platelets' ability to adhere and aggregate was monitored with the use of platelet function analyser (PFA-100 collagen/epinephrine closure time, CT(CEPI) or collagen/ADP closure time, CT(CADP)), classical turbidimetric aggregometry and whole blood electrical aggregometry (WBEA), using collagen (WBEA(coll)), ADP (WBEA(ADP)) and arachidonic acid (WBEA(AA)) as platelet agonists, in 48 control healthy volunteers (mean age+/-S.D., 49+/-9 years) and 31 type 2 DM patients (50+/-9 years; HbA(1c) 9.4+/ 1.6%). In majority of control subjects (69%) and minority of diabetic patients (29%, p=0.0006), the use of 150 mg aspirin daily for 1 week significantly reduced platelet adhesiveness and reactivity (by 14.1% in diabetes vs. 78.6% in control, p(np)=0.0035, as expressed by the relative changes in CT(CEPI)). Aspirin reduced WBEA(coll) and WBEA(AA) to a lesser extent in diabetic patients (by 2.1% vs. 8.3% in controls, p(np)=0.0397, and by 97.3+/-12.8% vs. 100% in controls, p(np)=0.0383, respectively), which corresponded to ASA-mediated decreased aggregation in platelet-rich plasma (PRP, r(S)=0.45 and r(S)=0.78 for collagen- or arachidonate-agonized platelets, p<0.01 or lower). The maximal inhibition of platelet aggregation was lower and IC(50) higher in diabetic compared to control subjects, both in the presence of arachidonic acid (71% vs. 39%, p(np)0.0001; 0.5 microg/ml vs. 1.3 microg/ml, p<0.0001) and collagen (52% vs. 35%, p<0.0004; 1.6 microg/ml vs. 2.1 microg/ml, p<0.01). The reduced response of platelets from diabetic subjects to aspirin was associated with a higher level of HbA(1c), lower concentration of HDL-cholesterol and a higher total cholesterol concentration. Overall, there is evidence that reduced platelets response to aspirin may occur more often in diabetic patients. Poor metabolic control may play a role in the reduced platelet sensitivity to aspirin in DM patients. Thus, our findings strongly support the requirements for an excellent near-normal metabolic control and may suggest a need for alternative ASA dosing schedules in DM patients. PMID- 15115667 TI - Thrombomodulin, von Willebrand factor and E-selectin as plasma markers of endothelial damage/dysfunction and activation in pregnancy induced hypertension. AB - OBJECTIVE: Endothelial disturbance (whether activation, dysfunction or damage) is a likely pathogenic mechanism in pre-eclampsia and pregnancy-induced hypertension (PIH). We set out to determine which of three plasma markers of endothelial disturbance, indicating endothelial activation (E-selectin) or damage/dysfunction (von Willebrand factor (vWf), soluble thrombomodulin), would provide the best discriminator of PIH compared to normotensive pregnancy. STUDY DESIGN: Cross sectional study of 36 consecutive women with PIH (age 31+/-6 years) and 36 consecutive women with normotensive pregnancies (age 29+/-5 years) of similar parity. Plasma levels of vWf, E-selectin and thrombomodulin were measured using ELISA. RESULTS: As expected, women with PIH had significantly higher levels of plasma vWf (by 19%, p=0.003), E-selectin (by 40%, p<0.001) and thrombomodulin (by 61%, p=0.01) than normotensive women. However, on stepwise multiple regression analysis, only thrombomodulin was an independent significant predictor of the presence of PIH (p=0.023). CONCLUSIONS: We conclude that although vWf, E-selectin and thrombomodulin are all raised in PIH, only thrombomodulin was independently associated with PIH. This molecule could potentially be useful in monitoring and in providing clues in aetiology and pathophysiology, and may have implications for the clinical complications associated with PIH. PMID- 15115666 TI - Point-of-care ecarin clotting time versus activated clotting time in correlation with bivalirudin concentration. AB - INTRODUCTION: A thrombin inhibitor management (TIM) point-of-care test based upon the ecarin clotting time (ECT) has been developed. The ECT has been suggested to more accurately reflect the anti-coagulant effect of direct thrombin inhibitors compared with the activated clotting time (ACT). We sought to examine the correlation of the TIM-ECT test with bivalirudin concentration in patients undergoing percutaneous coronary intervention (PCI), and to compare the performance of this test with the current standard (i.e., ACT). MATERIALS AND METHODS: In a multicenter study, blood samples were obtained at six pre-defined time-points in 170 consecutive patients undergoing PCI using bivalirudin. For each sample, the TIM-ECT (citrated and non-citrated), ACT, and bivalirudin concentration was determined. RESULTS: Considering samples from all time-points (n=784), the correlations of TIM-ECT citrated, TIM-ECT non-citrated, and ACT with bivalirudin concentration were 0.96, 0.93, and 0.90, respectively. For samples collected at therapeutic levels of bivalirudin (n=353), the correlations of TIM ECT citrated, TIM-ECT non-citrated, and ACT with bivalirudin concentration were lower, and showed a greater disparity between methods, with correlation coefficients of 0.75, 0.59, and 0.37, respectively. Prediction models based on the measured bivalirudin concentration were developed for TIM-ECT and ACT, and the coefficients of determination (r(2)) of actual versus predicted TIM-ECT and ACT were 0.91 and 0.81, respectively. CONCLUSIONS: In this PCI population, the TIM-ECT point-of-care test and ACT demonstrated a strong correlation with bivalirudin concentration. The TIM-ECT test had a higher correlation with bivalirudin concentration at therapeutic levels of the drug, and for individual samples appears to more consistently reflect the bivalirudin concentration compared with the ACT. PMID- 15115668 TI - The effects of arm cranking exercise and training on platelet aggregation in male spinal cord individuals. AB - Platelet aggregation at rest and in responses to exercise and training were compared between spinal cord injured (SCI) individuals (N=5) and able-bodied subjects (N=7). All participants performed arm cranking exercise at 60-65% VO(2peak) for 30 min. Venous blood samples were obtained before and after sub maximal exercise and measured for platelet aggregation using ADP and collagen. To assess the effects of arm cranking training, platelet aggregation was re-measured in all subjects at rest and in response to the sub-maximal arm cranking exercise after 12 weeks of individually supervised training programme. Before training, the resting mean values of platelet aggregation induced by ADP and collagen were not different (P>0.05) between SCI and able-bodied. However the SCI individuals, but not the able-bodied subjects, exhibited a significantly (P<0.05) higher maximal platelet aggregation induced by ADP and collagen following sub-maximal arm cranking exercise. Although VO(2peak) after training was significantly increased (P<0.05) in both groups, the resting mean values of platelet aggregation induced with ADP and collagen were not significantly different (P>0.05) from those observed before training and were not different (P>0.05) between SCI and able-bodied. Post-training, the SCI individuals, but not able bodied individuals, exhibited a significant decrease (P<0.05) in platelet aggregation following sub-maximal arm cranking exercise and this occurred with both ADP and collagen. These results suggest that SCI individuals, but not normal subjects increase their platelet aggregation following sub-maximal arm cranking exercise. Furthermore, arm cranking training in SCI individuals, appears to diminish the percentage of platelet aggregation ex vivo. PMID- 15115669 TI - The impact of peripheral arterial disease on circulating platelets. AB - INTRODUCTION: To test the hypothesis that circulating platelets display evidence of reversible interactions with atherosclerotic lesions, platelet alpha-granule content and propensity for microaggregate formation were measured in samples from normal donors (n=65) and from patients with either peripheral arterial disease (n=47) or renovascular hypertension (n=22). To measure the effect of a defined arterial injury on platelet function, platelet samples were compared before and 30 min after elective angioplasty. MATERIALS AND METHODS: P-selectin was measured after strong stimulation of ultra-dilute platelets with thrombin (10 nM). Microaggregation was measured as a platelet count deficit in citrate anticoagulated platelet-rich plasma (PRP) relative to that predicted from the count in EDTA-anticoagulated blood. RESULTS: Platelet alpha-granule P-selectin was significantly lower from platelets of patients compared to normal donors. In addition, platelets from patients have a significantly greater propensity to form microaggregates in citrate anticoagulant. In contrast to atherosclerotic renovascular hypertension, platelets from patients with fibromuscular dysplasia, a distinct non-inflammatory cause of arterial stenosis, do not differ significantly from normal donors. Other than the PRP platelet count, which rose transiently following angioplasty, other platelet measures were unchanged by the injury. CONCLUSIONS: Atherosclerotic arterial disease is associated with an increased share of platelets unable to express P-selectin and an increased fraction of platelets that microaggregate in citrate anticoagulant. These platelet alterations are not completely explained by either focal arterial injury or abnormal rheology associated with arterial stenosis but appear to be an effect of the atherosclerotic process. PMID- 15115670 TI - Lonofibrase, a novel alpha-fibrinogenase from Lonomia obliqua caterpillars. AB - Envenomation caused by Lonomia obliqua caterpillars is an increasing problem in Southern Brazil. The clinical profile is characterized by a profound hemorrhagic disorder. In the present study, we describe the characterization of a fibrin(ogen)olytic factor (lonofibrase) isolated from a venomous secretion of the caterpillars. The crude extract showed a dose-dependent inhibitory effect in the rate of thrombin-induced fibrinogen clotting and produced fragmentation of fibrinogen. Isolation of the fibrin(ogen)olytic enzyme was achieved by combining ion exchange chromatography followed by gel filtration in a fast protein liquid chromatography (FPLC) system. A single 35-kDa band was identified and the isolated enzyme named lonofibrase. Lonofibrase rapidly degrades Aalpha and Bbeta chains of fibrinogen, also being able to cleave fibrin in a distinct way from that observed with plasmin. The presence of lonofibrase with both fibrinogenolytic and fibrinolytic activities in L. obliqua secretion is coherent with the severe hemorrhagic clinical profile resulting from envenomation caused by these insects. PMID- 15115671 TI - The effect of anticoagulation with subcutaneously delivered polyethylene glycol conjugated hirudin and recombinant tissue plasminogen activator on recurrent stenosis in the rabbit double-balloon injury model. AB - Myointimal hyperplasia is the condition usually responsible for recurrent stenosis (restenosis) after endarterectomy, bypass grafting and angioplasty. Its cause is still not known. The present study examined whether inhibition of thrombin by tissue plasminogen activator (r-TPA) or polyethylene glycol recombinant hirudin (PEG-hirudin) could reduce restenosis in an animal model. Restenosis was induced in 20 cholesterol-fed rabbits. The right carotid artery underwent a double-balloon injury while left carotid artery acted as a control. Recombinant tissue plasminogen activator (1 mg kg(-1) s.c.) and PEG-hirudin (0.7 mg kg(-1) s.c.) were given subcutaneously with normal saline acting as a control. Blood levels of PEG-hirudin were measured by both ELISA and an Ecarin (activity) assay. Vessel dimensions were measured in histological sections, obtained from perfusion-fixed tissue, using computerised planimetry. The model reproduced many of the histological changes found in human restenosis, such as intramural thrombus, rupture of the elastic lamina, macrophage infiltration and smooth muscle migration. Reinjury caused an almost three-fold reduction in the area of the lumen (median 0.25 mm(2)) compared with uninjured vessels (median 0.72 mm(2)). The mean plasma levels of PEG-hirudin and r-tPA achieved were 291 ng/ml (S.E.M. 28 ng/ml) and 34 IU/ml (S.E.M. 12 IU/ml), respectively. PEG-hirudin significantly inhibited the effect of balloon injury on luminal area compared with saline-treated controls (0.21 versus 0.44 mm(2), respectively, P<0.05). Recombinant tPA also had a similar inhibitory affect, but this did not reach statistical significance (0.16 versus 0.44 mm(2), respectively, P>0.05). The magnitude of luminal narrowing was significantly reduced by subcutaneous injection of PEG-hirudin. Further studies are required to determine whether this effect can be enhanced by other antithrombins or improved methods of delivery. PMID- 15115672 TI - Heparin cofactor II is more sensitive than antithrombin to secretory impairment arising from mutations introduced into its carboxy-terminal region. AB - INTRODUCTION: Antithrombin (AT) and heparin cofactor II (HCII) are plasma glycoproteins and serpins that inhibit thrombin. We showed [Blood 86 (1995) 3461] that recombinant rabbit AT containing the Utah mutation of AT, P407L, was inefficiently secreted by transfected primate and rodent cultured cells. In the current study, the effects of P407L and related substitutions in human AT and human HCII were investigated. MATERIALS AND METHODS: Cultured cells were transfected transiently (COS-1) or permanently (CV-1) with AT and HCII expression vectors encoding the wild type or mutant serpins. The amount of protein secreted was determined immunologically, while RNA levels were assessed by reverse transcription-PCR (RT-PCR). The kinetics of secretion were investigated by pulse chase experiments, supplemented by endoglycosidase H or lactacystin treatment. RESULTS AND CONCLUSIONS: The F450L, P455L, P477L, P477*, and T446* (*=stop codon) mutations reduced HCII secretion 6.6- to 24-fold, while the F402L, A404T, and P407L mutations reduced AT secretion in COS-1 cells 1.7- to 5.2-fold. Homologous mutants HCII (P455L) and AT (P407L) were transcribed at similar levels in COS-1 cells, but were secreted less rapidly and less efficiently than their wild-type counterparts. HCII (P455L) exhibited intracellular proteasomal degradation in permanently transfected CV-1 cells, while AT (P407L) secretion was unaffected in this milieu. HCII secretion is thus more sensitive than that of AT to C-terminal mutations, as shown in two primate cell lines, likely reflecting a greater tendency to misfold during synthesis. We speculate that this difference may arise due to an interstrand s1C/s4B loop that is shorter in HCII than in AT. PMID- 15115673 TI - Trimming of peripherally inserted central venous catheters may increase the risk of thrombosis. PMID- 15115674 TI - Advances in periodontal disease markers. AB - A computer-assisted medline search was conducted to find the relevant articles concerning the periodontal disease markers in gingival crevicular fluid (GCF) and saliva published during the 10-year period from 1993 to July 2003. This review suggests that certain diagnostic uses of saliva and GCF show promise. Although both fluids have been used to evaluate the risk for an individual to develop periodontal disease and to monitor of the host response to periodontal therapy, GCF has the chance of being closely approximated to the periodontal tissues where periodontal disease begins. The enzymes contributed to extracellular matrix (ECM) molecules and non-ECM molecules degradation and markers for polymorphonuclear leukocytes (PMN) activity and influx into the gingival tissue seem to provide valuable information regarding the periodontal disease diagnosis and prognosis. There is also an increasing evidence implicating reactive oxygen species and nitric oxide pathway in the pathogenesis of periodontal diseases. Although promising results have been achieved with the assays evaluating the markers in assessment of periodontal disease status, up to now, none of these tests are used routinely. Further, one commercially available genetic test has been reported to have the potential to be used to predict the periodontal disease, but there are controversial reports on this genetic susceptibility test. PMID- 15115675 TI - Advances in recombinant antibody microarrays. AB - Antibody microarrays, one emerging class of proteomic technologies, have broad applications in proteome analysis, disease diagnostics and quantitative analysis. Compared to DNA microarrays, protein targets have significantly more complex interactions with their ligands such as antibodies. To introduce antibody microarrays for clinical diagnostics and thus to complement or replace conventional immunoassays, several new developments are addressed. We discuss different microarray surfaces, immobilization techniques, detection systems and advantages and disadvantages of antibody microarrays compared to standard clinical techniques. Currently, the probes with highest specificity, well characterized binding properties, and the possibility of large-scale production using display libraries are recombinant antibodies. PMID- 15115676 TI - Advances in understanding what we measure when detecting anticardiolipin autoantibodies. AB - Understanding so-called anticardiolipin autoantibodies has been the focus of significant interest due to their accepted role in the pathology of antiphospholipid syndrome (APS). How these antibodies contribute to thrombosis, stroke and recurrent fetal loss is not clearly understood. A precise determination of the antigenic epitope(s) recognized by anticardiolipin autoantibodies will contribute to an understanding of their role in this complex disease; however, there have been many conflicting reports regarding these epitope(s), and many of the apparent contradictions arise from the assay systems that have been used. In this review, we attempt to highlight what we believe are the salient points on this issue. We present further evidence supporting our hypothesis that most of these autoantibodies recognize epitope(s) located on domain 1 (DI) of beta2-glycoprotein 1 (beta2GPI). PMID- 15115678 TI - Biochemical analysis of pleural, peritoneal and pericardial effusions. AB - Body fluids other than blood, urine and cerebrospinal fluid are often submitted for biochemical analysis. Of these, pleural, peritoneal and pericardial fluids are the most common. Laboratory tests are a useful tool to assess the aetiology, pathophysiology and subsequent treatment of effusions. A wide range of biochemical tests may be requested. This review critically examines the various analytes that have been used to investigate these body fluids. PMID- 15115677 TI - Towards fast and inexpensive molecular diagnostic: the case of TP53. AB - BACKGROUND: Much research suggests that TP53 mutations have prognostic importance and sometimes are a significant factor in clinical oncology. A considerable effort has been made to develop fast and inexpensive methods for TP53 mutations detection. METHODS: On the basis of describing the role of TP53 as tumor suppressor gene and TP53 mutation spectrum, the authors discuss conventional methods and new technologies for TP53 mutations detection. This discussion is supported by more recent publications in the field of both molecular genetics and analysis technologies. RESULTS: Biosensors and gene chips are of considerable recent interest, due to their tremendous promise for obtaining sequence-specific information in a faster, simpler and cheaper manner compared to traditional methods. CONCLUSIONS: New methods such as biosensors and gene chips appear promising as analytical methods of detecting mutations. PMID- 15115679 TI - Differentiation of human umbilical cord blood CD133+ stem cells towards myelo monocytic lineage. AB - BACKGROUND: Characterisation of stem cells by flow cytometry, their expansion and differentiation are presently of major interest for cell engineering as the basis of a therapeutic concept for transplantation. Haematopoietic stem cells (HSC) express CD34, the adhesion structure which binds 2L-selectin, CD117, a receptor for stem cell factor (SCF; c-kit ligand), and CD133, a transmembrane protein belonging to the family of mucoproteins. METHODS: The aim of the present investigation was the systematic investigation of proliferation and differentiation characteristics of umbilical cord blood stem cells (UCBSC) isolated by an immmunomagnetic separation system using CD133 antibody-coated microbeads and to evaluate the effects of different sera and various concentrations, as well as the effects of IL-3 and IL-6 on total cell expansion and differentiation of isolated CD133+ cells. Differentiation patterns were measured by flow cytometry. RESULTS: After the immmunomagnetic separation the yield of CD133+ cells was 0.45+/-0.17 x 10(6) cells/ml; the purity of isolated CD133+ cells was 95.79+/-1.86%. The majority of CD133+ cells coexpressed CD117. The most pronounced expansion during cultivation of 2 weeks was achieved in media supplemented with 12.5% horse serum plus 12.5% fetal calf serum (FCS) with stem cell factor and interleukine 3; the fold-expansion was 16.67+/-6.20. During the cultivation period, UCBSC were constantly loosing stem cell markers and differentiated towards myelo-monocyte lineage (granulocytes and/or monocytes). CONCLUSIONS: These in vitro results demonstrate that thorough investigation of various cultivation conditions is needed for successful expansion and differentiation of stem cells towards different lineages to be used therapeutically for replacement of damaged cells. PMID- 15115680 TI - Validity of plasma remnant lipoproteins as surrogate markers of antemortem level in cases of sudden coronary death. AB - Hitherto triglycerides (TG) and TG-rich lipoproteins were been of limited value as surrogates for antemortem levels. We measured TG levels in postmortem plasma from sudden coronary death cases (SCD, n=91) by using two TG assays, Dry Chem TG (free glycerol was added) and the Determiner L-TG (without added free glycerol) that measured net TG. TG levels were markedly higher by the Dry Chem TG (y) vs. Determiner L-TG (x), y = 1.03x + 229 mg/dl. HPLC showed large amounts of free glycerol in postmortem plasma and in TG-rich lipoprotein remnants (RLP). These results were verified in a rabbit model of SCD. Further, RLP from SCD were found to be biophysically similar to those from living patients with coronary artery disease (CAD). In conclusion, postmortem plasma sampled up to 12 h after death is appropriate for measuring lipid and lipoproteins, TG and RLP-TG as surrogates for antemortem levels when a TG assay without added free glycerol is used. PMID- 15115681 TI - Improving homocysteine levels through balneotherapy: effects of sulphur baths. AB - BACKGROUND: Plasma homocysteine (tHcy) is a risk factor for cardio-vascular diseases. Furthermore it has been associated with antioxidative status. Additionally balneotherapeutic sulphur baths have been shown to influence antioxidative status. METHODS: 40 patients with degenerative osteoarthrosis were randomised into two equal groups, a treatment group, receiving stationary spa therapy plus daily sulphur baths (sulphur group) and a control group receiving spa therapy alone (control group). Blood tHcy levels and urinary 8-OHdG (an indicator for oxidative stress) were measured at the beginning and the end of spa therapy. RESULTS: tHcy (micromol/l) was significantly reduced from 11.41 (+/ 2.91) to 10.55 (+/-2.28) in the sulphur group (p=0.016) and rose insignificantly from 12.93 (+/-2.28) to 13.80 (+/-3.87) in the control group. 8-OHdG (ng 8 OHdG/mg creatinine) declined from 18.00 (+/-18.28) to 11.16 (+/-5.33) in the sulphur group (n.s.) and from 17.91 (+/-5.87) to 18.17 (+/-5.70) in the control group (n.s.). Differences between the two groups showed significant effects of sulphur baths for tHcy (p=0.006) but not for 8-OHdG (p=0.106). CONCLUSIONS: Sulphur baths exert beneficial effects on plasma tHcyt whereas effects on 8-OHdG seem to be unlikely. PMID- 15115682 TI - A microtitre format assay for proline in human serum or plasma. AB - BACKGROUND: Recent studies have suggested that low serum proline concentration may be associated with low bone mineral density. However, further investigation of this association has been hampered by the lack of a relatively high throughput assay for proline in biological fluids. Here we report a sensitive and specific microtitre plate format assay for proline which exploits the chemical interaction between proline and isatin. METHODS: Human serum or plasma is deproteinised by incubation with sodium citrate buffer pH 4.1 at 95 degrees C, and the supernatant is reacted with isatin at 95 degrees C for 3 h. The resultant blue coloured product is quantitated sprectrophometrically. RESULTS: This assay yields a linear standard curve in the range 15 micromol/l to 1 mmol/l (r=0.998+/-0.002; n=8 determinations) with a sensitivity of 31+/-11 micromol/l. None of the other proteogenic amino acids are detected (<0.3% detection at 10 mmol/l) and the closely related metabolite hydroxyproline is only very weakly detected (3% detection at 10 mmol/l). Using human serum, the assay has linear dilution characteristics and a mean spike recovery of 107+/-5%. Repeated re-measurement of the same serum sample yields an intra-assay coefficient of variation (CV) of 4.8% and an inter-assay CV of 6.1%. CONCLUSIONS: This method provides the first reliable micro-titre format assay for proline in human serum. PMID- 15115683 TI - Maldi-TOF analysis of portal sera of pancreatic cancer patients: identification of diabetogenic and antidiabetogenic peptides. AB - BACKGROUND: Pancreatic cancer (PC) associated diabetes mellitus (DM) might be consequent to the diabetogenic effects of tumour products, possibly acting via nitric oxide (NO). Our aims were: (1) to verify whether PC associated DM determines an increased hepatic NO and (2) using MALDI-TOF analysis, to evaluate the peptide composition of PC cell conditioned media (CM) and of portal sera from patients with PC with (n=7) or without (n=4) DM. METHODS: In liver tissue homogenates of 23 patients with PC (n=17) or chronic pancreatitis (n=6) GAPDH mRNA and activity, glucose, lactate, nitrite and nitrate were assayed. MALDI-TOF analysis was performed in three PC cell lines CM, and in portal sera from patients with PC. RESULTS: Higher GAPDH mRNA and nitrite were found in patients with than in patients without DM. In PC cell CM, only 9 among a total of 75 fragments identified, were tumour specific. One hundred seventy-three fragments were identified in the portal sera of patients: one was positively and six fragments were negatively correlated with DM. CONCLUSIONS: Unlike liver GAPDH, NO appears to be involved in PC associated DM. In portal sera, the absence, rather than the presence, of specific fragments, appears to be correlated with the development of DM. PMID- 15115684 TI - Multiplex allele-specific PCR assay for differential diagnosis of Hb S, Hb D Punjab and Hb Tak. AB - BACKGROUND: Apart from hemoglobin (Hb) E, Hb D-Punjab [beta121(GH4)Glu-Gln] and Hb Tak [beta147Term-Thr] are the two most common beta-chain variants among the Asian population. These two Hb variants have similar alkaline electrophoretic mobilities and HPLC profiles as those of the Hb S [beta6(A3)Glu-Val]. Differential diagnosis of these clinically relevant hemoglobinopathies is therefore problematic. Direct detection of the beta-globin gene mutations would be another diagnostic alternative. METHODS: A simultaneous DNA diagnosis of the three Hb variants was developed based on the multiplex allele-specific polymerase chain reaction (PCR) approach. The method was validated on 10 carriers of Hb D Punjab, 5 carriers of Hb Tak, 2 carriers of Hb S and 50 normal individuals of Thai origin. RESULTS: The three abnormal Hbs could be correctly diagnosed with the simultaneous PCR approach, and a complete concordance with results using other established methods was obtained. CONCLUSIONS: The multiplex allele specific PCR approach developed should prove useful in complementing routine Hb analysis for differential diagnosis of these three common Hb variants and should facilitate a program of hemoglobinopathy screening in the region. PMID- 15115685 TI - Endotoxin sensitivity and immune competence in chronic heart failure. AB - BACKGROUND: Raised concentrations of endotoxin (lipopolysaccharide, LPS) are found in patients with chronic heart failure (CHF). Tolerance of monocytes to LPS can be induced by LPS itself resulting in a downregulation of cytokine response to LPS challenge. This phenomenon of LPS desensitization has also been suggested for CHF. METHODS: We investigated whether CHF patients really show a desensitization to LPS stimuli at rest or after physical exercise, which was used as a model of limited inflammatory reaction. Thirty-five patients with CHF (59+/ 12 years, 8 women) and 30 healthy control subjects were prospectively studied with cardiopulmonary exercise testing. At rest and directly after exercise blood samples were taken for the quantitative determination of HLA-DR expression of monocytes as a measure for immune competence and for the measurement of TNFalpha generation after ex vivo stimulation by LPS. RESULTS: HLA-DR expression was comparable in CHF patients and controls at rest as well as after exercise. TNFalpha production by LPS-stimulated monocytes ex vivo was higher in CHF patients compared to controls at rest and after exercise. CONCLUSIONS: Thus, our data are the first to show that patients with stable CHF show a cellular hypersensitivity to LPS with a higher TNFalpha generation capacity at rest and after exercise compared to controls. CHF patients seem to have a marked susceptibility to low inflammatory stimuli and no desensitization to LPS. PMID- 15115686 TI - The effects of thinner inhalation on superoxide dismutase activities, malondialdehyde and glutathione levels in rat lungs. AB - BACKGROUND: Recent years' usage of thinner by the young generation as a drug constitutes a serious problem in the society. Due to common usage in the industrial sector, most people are affected from the manufacturing process to the consuming phase. AIM: Because of these reasons, this project has been preferred to research the effects of thinner on oxidant and antioxidant status. METHODS: Totally 46 rats were included in the study. Thirty six rats were separated into six groups with 10 rats in a control group. The first group inhaled thinner for 2 weeks, and the other groups were exposed to thinner for 4, 6, 8, 10 and 12 weeks for 1 h twice a day. On the mentioned duration, rats were autopsied. Lung tissues malondialdehyde (MDA), glutathione (GSH) levels and superoxide dismutase (SOD) activities were determined to designate the oxidant-antioxidant balance. RESULTS: We observed an increase in MDA values both in the acute and the subacute periods. In the chronic period by the consuming of lipid peroxidation products, MDA values decreased and as the oxidative stress continued MDA values again increased. We observed that especially GSH values that has antioxidant feature, decreased until 6 weeks in order to compensate lipid peroxidation products. In the consuming period of lipid peroxidation, the values became fixed and later, these values again increased. There was no relationship between the changing values of MDA and SOD. CONCLUSIONS: Thinner is an agent that causes oxidative stress and inhalation of high doses of thinner causes harm to the respiratory system. As there are few reports in the literature on long-term effects of thinner inhalation, more studies might be necessary. PMID- 15115687 TI - Biochemical study on beta-glucosidase in individuals with Gaucher's disease and normal subjects. AB - BACKGROUND: Gaucher's disease (GD) is a disorder caused by the deficiency of lysosomal beta-glucosidase, an enzyme that participates in the degradation of glycosphingolipids. Deficiency of this enzyme results in the accumulation of glucocerebrosides in macrophage lysosomes. No studies comparing the biochemical and kinetic behavior of this enzyme in leukocytes and fibroblasts from normal individuals and patients with Gaucher's disease are available. METHODS: We compared the activities of beta-glu and chitotriosidase between normal subjects and Gaucher disease patients, and characterized the behavior of beta-glu in terms of pH optimum, heat stability, Km and Vmax. RESULTS: The results showed a different behavior of the enzyme in the groups analyzed. CONCLUSIONS: This finding might be useful in cases in which the measurement of enzyme activity alone is not reliable for the establishment of the diagnosis of Gaucher's disease. PMID- 15115688 TI - The effect of elevated serum soluble CD40 ligand on the prognostic value in patients with acute coronary syndromes. AB - BACKGROUND: Increasing evidence shows that high expression of CD40L plays an important role in the pathogenesis of atherosclerosis and coronary artery disease. We evaluated the clinical predictive value of increased serum soluble CD40 ligand (CD40L) in patients with acute coronary syndromes (ACS) and acute chest pain. METHODS: Serum levels of soluble CD40 ligand were measured by ELISA in 128 patients with ACS and in 68 patients with acute chest pain. Platelet activation was assessed by flow cytometry. RESULTS: The levels of soluble CD40 ligand were increased in 57.8% patients with ACS (>8.0 ng/ml) and in 35 patients with acute chest pain (>8.0 ng/ml), respectively. The level of soluble CD40 ligand was slightly correlated with measured levels of troponin T (r=0.21, p<0.05), and the increased soluble CD40L levels (>8.0 ng/ml) were associated with higher risk for AMI, sudden death and recurrent angina. Patients with elevated serum levels of sCD40L and cTnT showed a significantly increased risk of major adverse cardiovascular events (including AMI, sudden death and recurrent angina) in the two groups during 30 days and 6 months of follow-up. CONCLUSION: In patients with unstable coronary artery disease, elevation of serum soluble CD40L levels indicated an independent increased risk of major adverse cardiovascular events. PMID- 15115689 TI - Cytokine production by non-adherent mouse splenocyte cultures to Echinacea extracts. AB - BACKGROUND: Echinacea is commonly used in oral dosage as an immune stimulant to increase resistance to viral, bacterial and fungal infections of the upper respiratory tract. It has been suggested that Echinacea is able to stimulate innate immune responses, including those regulated by macrophages and natural killer cells. Indeed, macrophages respond to purified polysaccharide and alkylamide preparations. However, the mechanisms for stimulation of cells responsible for adaptive immunity have not been fully elucidated for other molecules present in Echinacea purpurea preparations. METHODS: Adherent and non adherent mouse splenocyte populations were incubated in vitro with Echinacea, or with water or alcohol soluble Echinacea extract preparations. Supernatants were collected at 48-h post-incubation, and tested by standard ELISA for presence of secreted cytokines and proinflammatory mediators. RESULTS: Whole splenocyte populations were capable of producing significant amounts IL-6 (1014 pg/ml) in response to Echinacea preparations. The response was primarily contained towards products isolated to the water extract preparation; no IL-6 was produced upon challenge with the alcohol extract. The IL-6 response was produced by the non adherent cellular population, which made 4912 pg/ml IL-6 when treated with water soluble extract at 1 mg/ml. Likewise, the water soluble extract of Echinacea was able to stimulate non-adherent splenocyte populations to produce TNF-alpha (2082 pg/ml), IL-10 (892 pg/ml) and MIP-1alpha (6486 pg/ml) from non-adherent splenocytes, but only significant concentrations of TNF-alpha and MIP-1alpha mediators were produced from adherent populations at similar dose concentrations. Neither population of splenocytes was capable of stimulating significant production of IFN-gamma, IL-2 or IL-12 to any preparation of Echinacea examined. CONCLUSIONS: The immune stimulatory ability of components contained within E. purpurea extracts offer insight into possible therapeutic potential of this product to regulate non-adherent lymphocytes in immune responses and activation events. PMID- 15115690 TI - An automated 'bio-intact' PTH assay: a step towards standardisation and improved correlation with parathyroid function in renal disease. AB - BACKGROUND: Most methods for the determination of parathyroid hormone (PTH) show cross-reactivity with N-truncated forms of PTH. The analytic and diagnostic value of a recently developed automated PTH test without this cross-reactivity was examined. METHODS: The PTH levels of 73 patients undergoing hemodialysis were compared using the 'bio-intact' PTH (Nichols Institute Diagnostics) and 3 'intact' PTH tests (from Nichols, Roche Elecsys and Diagnostics Corporation DPC). Further, the (non 1-84) PTH fragment and the PTH ratio (bio-intact PTH/(non 1-84) PTH) were calculated. All results were then correlated with biochemical bone markers (bone-specific alkaline phosphatase and bone collagen C-terminal telopeptides in serum). RESULTS: 'Bio-intact' PTH values were lower than the PTH results generated by the 'intact' PTH assays. Results of all PTH tests were closely correlated (r=0.96-0.98, p<0.01). Correlations with biochemical bone markers were high (r=0.31-0.63, p<0.01), but no significant association between the PTH ratio and all other tests (r=-0.2 to 0.03) was found. CONCLUSIONS: In stable hemodialysis patients, the different PTH tests show a similar correlation with the bone markers. It is however desirable to measure PTH with assays devoid of any cross-reaction for a better comparability. In this study, the PTH ratio was not correlated with biochemical bone markers; the use of this ratio requires further investigation. PMID- 15115691 TI - Cell cycle analysis of the CD133+ and CD133- cells isolated from umbilical cord blood. AB - BACKGROUND: Umbilical cord blood cells (stem/progenitor cells) exhibit high proliferative capacities leading to a large expansion of cells in appropriate cell culture conditions. The aim of this study was to evaluate by flow cytometry the cycling status of CD133+ and CD133- cells depending on various culture conditions, such as sera, stem cell factor (SCF), interleukin 3 (IL-3) and interleukin 6 (IL-6). METHODS: An immunomagnetic system was used for cell separation. CD133+ and CD133- cells were seeded in Iscove's Modified Dulbecco's Medium (IMDM) with different serum concentrations and were stimulated with SCF (100 ng/ml), IL-3 (50 ng/ml) and IL-6 (50 ng/ml). RESULTS: Our experiments demonstrated that immediately after separation, 96.75+/-0.58% of CD133+ cells and 97.04+/-1.76% of CD133- cells were in G0/G1-phase, while 2.02+/-0.38% and 0.88+/ 0.52% were in the S-phase, respectively. Our data documented that CD133+ cells are more active than CD133- cells after the first week of cultivation (p<0.01). Statistically significant difference was found for CD133+ cells vs. CD133- cells after second week of cultivation in G0/G1- and S-phases under all tested conditions. A combination of 12.5% FCS+12.5% HS yielded the highest cell expansion for CD133+ cells; this was concomitant with highest percentage of S phase and G2M-phase. Our data show that the medium with 25% HS was the best for cell expansion and cycling of the CD133- cells for the first week, followed by the 12.5% FCS+12.5% HS. After 2 weeks of cultivation, obviously 12.5% HS and 12.5% FCS+12.5% HS exhibited similar S-phase amounts in CD133- cells. A decrease of HS concentrations seemed to stimulate CD133- cells' S-phase after the second week. CONCLUSIONS: Our data indicate that the source and the concentration of the serum used for cultivation have an impact on both cell populations: CD133+ cells are most comfortable with a combination of FCS and HS; CD133- cells prefer media containing HS. Cell cycle status may be an important factor for defining cultivation strategies for stem cell expansion. PMID- 15115692 TI - Missense mutation W86R in exon 3 of the lipoprotein lipase gene in a boy with chylomicronemia. AB - BACKGROUND: Familial LPL deficiency is a rare inborn error of metabolism caused by mutational change within the LPL gene, which leads to massive hypertriglyceridemia. METHODS: The underlying molecular defect in a boy of Croatian descent was studied by SSCP analysis, DNA sequencing and finally confirmed by RFLP. RESULTS: DNA analysis showed the child to be a homozygote and his parents heterozygotes for TGG-->CGG change in codon 86 of the LPL gene, which leads to W86R amino acid substitution. DNA sequence analysis also showed a silent mutation in the third exon of father's DNA, V108V. Determination of some LPL gene polymorphisms showed the child and his parents to have HindIII/H+H+ and both S447 wild-type alleles, whereas for PvuII the parents had P(+)P- and the child P(+)P+ genotype. CONCLUSIONS: In this case, W86R mutation was the reason for the production of nonfunctional enzyme and consequently triacylglycerol (TG) exceeding 15 mmol/l. This implies the risk of frequent episodes of acute pancreatitis. Decreased LPL activity leads to elevated triacylglycerol levels and reduced HDL-cholesterol, both risk factors for the development of coronary artery disease. LPL genotyping especially of young patients with hypertriglyceridemia is therefore necessary and justifiable. PMID- 15115693 TI - Isotope dilution tandem mass spectrometric method for T4/T3. AB - BACKGROUND: The thyroid hormones thyroxine (T4) and 3,5,3'-triidothyronine (T3) are essential for regulating a number of biological processes, including growth, neurodevelopment, carbohydrate metabolism, oxygen consumption and protein synthesis. Immunoassays are the current methods for thyroid hormone measurement and suffer from a lack of specificity. Our objective was to simultaneously measure T4 and T3 using isotope dilution tandem mass spectrometry within a single run. To compare the results obtained by this MS/MS method with those obtained by an immunoassay procedure on the same samples (DPC Immulite for T3, Diagnostics Product, and Dade RxL Dimension for T4, Dade-Behring). METHODS: An API-3000 tandem mass spectrometer (SCIEX, Toronto, Canada) equipped with TurboIonSpray and Shimadzu HPLC system was used employing isotope dilution with deuterium-labeled internal standard (l-thyroxin-d2). The method requires 100 microl of serum and involves addition of internal standard, precipitation of proteins with methanol and injection of the supernatant onto a C-18 column. After washing, the switch valve is activated and T4 and T3 eluted using a methanol gradient. T4 and T3 by immunoassay were performed using the Dade RxL Dimension and the DPC Immulite, respectively. RESULTS AND CONCLUSIONS: An isotope dilution tandem mass spectrometry method for the simultaneous determination of total T4 and T3 in serum is described which is accurate, specific, precise (%CVs 3.5-9.0), simple and fast (<7 min). PMID- 15115694 TI - Effect of storage conditions on the extraction of PCR-quality genomic DNA from saliva. AB - BACKGROUND: Saliva is a potentially useful source of genomic DNA for genetic studies since it can be collected in a painless and non-invasive manner. We sought to determine whether different storage conditions of saliva samples impact our ability to extract genomic DNA that is of sufficient quality for use in the polymerase chain reaction (PCR). METHODS: Saliva was collected from healthy volunteers and 2-ml aliquots subjected to different storage conditions: S1- washing of saliva using phosphate-buffered saline (PBS) and extraction of DNA on the same day of collection; S2--washing and centrifugation to yield a pellet, which was stored at-70 degrees C for 1 week prior to DNA extraction; S3--storage of whole saliva at 4 degrees C for 7 days, followed by washing and extraction of DNA; S4--storage at 4 degrees C for 7 days, followed by washing and pellet formation. The pellet was stored at -70 degrees C for 1 month before extraction of the DNA; S5--storage at-70 degrees C for 1 month, followed by washing and extraction of DNA. DNA yield and purity was determined by spectrophotometry at 260 and 280 nm. Twenty nanograms of genomic DNA was used for the polymerase chain reaction, and the resulting PCR band was captured by digital photography and quantified. RESULTS: The amounts of DNA extracted from 2 ml of saliva varied widely under the different storage conditions, while purity of the DNA extraction, based on OD(260/280) ratios, was good and comparable. PCR resulted in the presence of a single specific product of the correct size from all samples regardless of saliva storage conditions. Quantification of PCR bands showed significant differences between the various storage conditions (P<0.05). Compared to S1 samples, PCR bands from conditions S2 and S3 were not as strong, while those amplified from S4 and S5 samples were the weakest. Post-hoc analyses showed that the means for conditions S4 and S5 were significantly different from S1-S3. Qualitatively similar results were obtained when the PCR experiment was repeated. CONCLUSIONS: Saliva can act as a useful source of genomic DNA, even when stored under less than optimal conditions. PMID- 15115695 TI - A sensitive enzymatic assay for the determination of sucrose in serum and urine. AB - BACKGROUND: Sucrose permeability has been suggested as a simple and non-invasive marker of gastric mucosal damage. We here report on a sensitive enzymatic assay using four sequential enzyme reactions coupled with reduced thio-NADPH. METHODS: Sucrose is phosphorylated by sucrose phosphorylase (EC2.4.1.7). The subsequent reaction in the presence of phosphoglucomutase (EC5.4.2.2) and glucose-1,6 diphosphate forms glucose-6-phosphate. Sucrose of the monad forms the dyad thio NADPH. The reaction is monitored by changes in absorbance at 405 nm. RESULTS: The lower limit of detection (3SD method) was 2.8 micromol/l for serum and 7.0 micromol/l for urine. The precision of the method was <4.0%, and has sufficient analytical range. CONCLUSIONS: The assay was sensitive enough to monitor serum sucrose concentrations during the sucrose permeability test and an automated assay may be useful in a large number of subjects. PMID- 15115696 TI - T13M mutation of lecithin-cholesterol acyltransferase gene causes fish-eye disease. AB - BACKGROUND: Lecithin-cholesterol acyltransferase (LCAT) esterifies free cholesterol (FC) in plasma and plays a crucial role in the maturation of prebeta1 HDL (lipid-poor HDL) into alpha-migrating HDL (spherical HDL). Natural mutations of LCAT gene cause familial LCAT deficiency (FLD) or fish-eye disease (FED). The relationship between mutations and their phenotypes gives important clues to the functions of specific regions of LCAT. We investigated the first homozygous case with a substitution of threonine to methionine at codon 13 (T13M) of LCAT gene. METHODS: We evaluated LCAT activity, LCAT distribution among HDL subfractions and conversion of prebeta1-HDL to alpha-migrating HDL by native two-dimensional gel electrophoresis (N-2DGE). RESULTS: The proband had corneal opacity, severe hypo alpha-lipoproteinemia, half-normal LCAT activity and near normal cholesteryl ester/total cholesterol (TC) ratio in plasma. These features were characteristic of FED. Plasma prebeta1-HDL concentration was near normal, but not converted to alpha-migrating HDL during 37 degrees C incubation. As expected, alpha-migrating HDL (especially large particles) was markedly reduced. In the immunoblot against LCAT, the small alpha-migrating HDL from the proband had much less LCAT in this patient than in controls. CONCLUSION: T13M mutation of LCAT gene causes FED. PMID- 15115697 TI - 14C-propionate incorporation assay by rapid filtration in multiwell plates. AB - BACKGROUND: Methylmalonic aciduria (MMA) is divided into two groups according to cobalamin dependency, and this classification is important for treatment. Unfortunately, there has been no rapid and reliable method for the evaluation of cobalamin dependency. METHODS: [14C]-propionate incorporation into intact cells in the presence of either media alone or media containing various amounts of cobalamin was measured using a 96-well filtration plate and vacuum manifold. Incorporation of radioactivity was measured by direct microplate scintillation. RESULTS: Using peripheral white blood cells from normal individuals, we obtained a linear relationship between the rate of 14C-propionate incorporation and the number of cells over a broad range (10,000 to 100,000 cells/well). 14C-propionate incorporation in cells from eight patients was 1% to 13% of parallel controls. CONCLUSIONS: In this report, we describe a rapid, sensitive and reliable method for evaluating the cobalamin dependency of methylmalonic aciduria. PMID- 15115699 TI - Evaluation of a glucose meter against analytical quality specifications for hospital use. AB - BACKGROUND: The value of glucose meters in point of care testing (POCT) by medical professionals and self monitoring of blood glucose (SMBG) by patients is well established. We evaluated the SureSteppFlexx glucose meter against objective targets for imprecision and total error (TE). METHODS: The SureStepFlexx blood glucose system uses a reflectance-based glucose oxidase (GO) method and reports plasma-equivalent glucose values. The reference method was the Beckman LX20 Pro glucose oxidase/oxygen electrode method. Patient samples and commercial aqueous, quality control (QC) material were used to assess imprecision. To determine total error of the meters, results obtained on patient heparinized blood were compared against results obtained by the reference method using plasma. Analyses were carried out by an experienced nurse and technologist. RESULTS: Both operators achieved imprecision of < or =5% for all measurements. Overall, the percentage of results deviating from the total error targets were 0-21% and 4-13% for the nurse and technologist, respectively. Compared with earlier studies, the percentages of reported results outside the < or =10% and < or =5% American Diabetes Association (ADA) criteria are significantly lower. CONCLUSIONS: The SureSteppFlexx glucose meter meets analytical quality requirements and is suitable for POCT use in our hospital. We propose a tiered approach and suggest minimum, desirable and optimum total error targets for glucose meters of < or =5%, 7.9% and 13%, respectively. PMID- 15115698 TI - Arg257Cys polymorphism of CYP2A13 in a Chinese population. AB - BACKGROUND: Human cytochrome P450 2A13 (CYP2A13) is involved in the activation of numerous toxicants and carcinogens, especially in the metabolic activation of 4 (methyl-nitrosamino)-1-(3-pyridyl)-1-butanone (NNK), a major tobacco-specific carcinogen. A functionally significant coding single nucleotide polymorphism (C3375T) in exon 5 of CYP2A13, which results in an amino acid substitution of Arg 257 to Cys, has been recently reported to exist in White, Black, Hispanic, and Asian individuals, with the variant 3375T allele frequencies being 1.9%, 14.4%, 5.8% and 7.7%, respectively. Since genetic background differs between ethnic groups, our present study aims to characterize the CYP2A13 Arg257Cys polymorphism in Chinese. METHODS: 258 healthy Chinese Han volunteers were involved in this study. Polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) assay was employed to genotype for the Arg257Cys polymorphism. RESULTS: Of all the 258 subjects, 27 (10.5%) heterozygotes and 1 (0.4%) homozygote for the 257Cys allele were detected. The frequency of the variant 257Cys allele in this Chinese population was 5.6% (95%CI: 4.2-7.0%). CONCLUSION: The CYP2A13 Arg257Cys variant represents a common polymorphism in Chinese, with the 257Cys allele frequency being similar to the Hispanic and Asian groups, but significantly lower than the Black. PMID- 15115700 TI - Minor elevations in troponin T values enhance risk assessment in emergency department patients with suspected myocardial ischemia: analysis of novel troponin T cut-off values. AB - BACKGROUND: A consensus document developed by a joint committee of the European Society of Cardiology and the American College of Cardiology redefines myocardial infarction (MI) using an increase of troponin I or T as compared to a reference control population (i.e., troponin T (TnT) of 0.01 microg/l). A clinical problem arises when an arbitrary cut-off point is selected for determination of MI (i.e., TnT> or =0.1 microg/l), as minor elevations of troponin are associated with increased cardiovascular risk in selected patients with acute coronary syndromes. METHODS: We prospectively studied 420 unselected patients being evaluated for suspected myocardial ischemia in the emergency department (ED). We compared a 99th percentile MI cut-off limit for TnT, determined by constructing a standard receiver operator curve from our ED population in whom an acute coronary syndrome was excluded, to a standard MI cut-off limit of 0.1 microg/l in assessing cardiovascular risk. We also assessed the prognostic value of detectable TnT concentrations below this 99th percentile MI cut-off, but above the upper reference limit of healthy controls. RESULTS: The diagnosis of acute coronary syndromes (ACS) was more frequent in groups with higher TnT concentrations: 16.8% with a normal TnT (<0.03 microg/l), 29.5% with detectable TnT below the 99th percentile MI limit (0.03-0.066 microg/l), 64.3% with detectable TnT between the 99th percentile and standard MI cut-offs (0.067-0.099 microg/l), and 85.4% with TnT> or =0.1 microg/l (p<0.001 for the trend). Thirty-day cardiovascular event rates increased for any detectable concentration of troponin: 1.3% with normal TnT, 4.8% with detectable TnT below the 99th percentile MI limit, 15.4% with TnT between the 99th percentile and standard MI cut-off limits, and 12.5% with TnT> or =0.1 microg/l (p<0.01 for the trend). CONCLUSION: Using an MI cut-off concentration for TnT from a "non-ACS reference" improves risk stratification, but fails to detect a positive TnT in 11.7% of subjects with an acute coronary syndrome. PMID- 15115701 TI - Effect of high bicarbonate hemodialysis on ionized calcium and risk of metastatic calcification. AB - BACKGROUND: Disturbances in calcium and phosphate metabolism among chronic hemodialysis patients result in renal osteodystrophy and vascular calcification. Even though it is the ionized fraction of calcium that is metabolically active, this measurement is generally not available and decisions are made on the basis of total calcium. Formulae to predict ionized calcium concentrations are available. METHODS: The OPTI Critical Care Analyzer with E-Ca cuvettes was used on-site to measure acid-base parameters, electrolytes, and ionized calcium. Additional assays included total calcium, phosphate, and albumin. RESULTS: Using a dialysate with 1.25 or 1.5 mmol/l calcium and 40 mmol/l bicarbonate, we observed a statistically significant increase in pH and total CO2 concentrations in post-dialysis blood. Total and ionized calcium increased significantly only in the patients with central venous catheters but not in those with fistulas or grafts. All patients experienced a decrease in phosphate concentrations. CONCLUSIONS: The metabolic alkalosis induced by high bicarbonate dialysate was not associated with a decrease in ionized calcium or a change in the calculated concentration product ratio for hydroxyapatite formation in the immediate post dialysis period. However, if a 40% phosphate rebound were to occur 2 h after termination of dialysis, the calculated risk of metastatic calcification would increase 2.8-fold compared to pre-dialysis conditions. Formulae to calculate ionized calcium are not useful in this population. PMID- 15115702 TI - Relationships between platelets and inflammatory markers in rheumatoid arthritis. AB - AIM OF THE STUDY: To investigate platelets and different inflammatory markers in conjunction with a substantial inflammatory reaction. We used individuals with active rheumatoid arthritis (RA) as an experimental cohort. METHODS: We selected 16 patients with active RA having at least one affected joint. On day 1, platelet and neutrophil counts together with C-reactive protein (CRP) were determined. We further analysed platelet volume (MPV) and plasma levels of thrombopoietin (TPO), P-selectin, myeloperoxidase and interleukin 6 (IL-6). After 2 years when all patients failed to show any swollen joints all analyses were repeated. RESULTS AND CONCLUSIONS: As expected platelet count, CRP and IL-6 were elevated in active RA. The measures correlated with each other thus reflecting the same characteristic of the inflammatory response. The neutrophil count, MPV and myeloperoxidase also mirror disease activity. They failed to correlate with other activity markers thus providing unique information. MPV and myeloperoxidase on day 1 correlated with recovery values. Therefore, they could be suitable to use when following the inflammatory reaction over a long period of time. PMID- 15115703 TI - Reflex testing III: efficient use of laboratory markers for anemia. PMID- 15115704 TI - Effect of chronically elevated CO2 on CA1 neuronal excitability. AB - To study the effect of chronically elevated CO(2) on the excitability and function of neurons, we exposed mice to 7.5-8% CO(2) for approximately 2 wk (starting at 2 days of age) and examined the properties of freshly dissociated hippocampal neurons. Neurons from control mice (CON) and from mice exposed to chronically elevated CO(2) had similar resting membrane potentials and input resistances. CO(2)-exposed neurons, however, had a lower rheobase and a higher Na(+) current density (580 +/- 73 pA/pF; n = 27 neurons studied) than did CON neurons (280 +/- 51 pA/pF, n = 34; P < 0.01). In addition, the conductance voltage curve was shifted in a more negative direction in CO(2)-exposed than in CON neurons (midpoint of the curve was -46 +/- 3 mV for CO(2) exposed and -34 +/- 3 mV for CON, P < 0.01), while the steady-state inactivation curve was shifted in a more positive direction in CO(2)-exposed than in CON neurons (midpoint of the curve was -59 +/- 2 mV for CO(2) exposed and -68 +/- 3 mV for CON, P < 0.01). The time constant for deactivation at -100 mV was much smaller in CO(2)-exposed than in CON neurons (0.8 +/- 0.1 ms for CO(2) exposed and 1.9 +/- 0.3 ms for CON, P < 0.01). Immunoblotting for Na(+) channel proteins (subtypes I, II, and III) was performed on the hippocampus. Our data indicate that Na(+) channel subtype I, rather than subtype II or III, was significantly increased (43%, n = 4; P < 0.05) in the hippocampi of CO(2)-exposed mice. We conclude that in mice exposed to elevated CO(2), 1) increased neuronal excitability is due to alterations in Na(+) current and Na(+) channel characteristics, and 2) the upregulation of Na(+) channel subtype I contributes, at least in part, to the increase in Na(+) current density. PMID- 15115705 TI - Modulatory effects of acid-sensing ion channels on action potential generation in hippocampal neurons. AB - Extracellular acidification has been shown to generate action potentials (APs) in several types of neurons. In this study, we investigated the role of acid-sensing ion channels (ASICs) in acid-induced AP generation in brain neurons. ASICs are neuronal Na(+) channels that belong to the epithelial Na(+) channel/degenerin family and are transiently activated by a rapid drop in extracellular pH. We compared the pharmacological and biophysical properties of acid-induced AP generation with those of ASIC currents in cultured hippocampal neurons. Our results show that acid-induced AP generation in these neurons is essentially due to ASIC activation. We demonstrate for the first time that the probability of inducing APs correlates with current entry through ASICs. We also show that ASIC activation in combination with other excitatory stimuli can either facilitate AP generation or inhibit AP bursts, depending on the conditions. ASIC-mediated generation and modulation of APs can be induced by extracellular pH changes from 7.4 to slightly <7. Such local extracellular pH values may be reached by pH fluctuations due to normal neuronal activity. Furthermore, in the plasma membrane, ASICs are localized in close proximity to voltage-gated Na(+) and K(+) channels, providing the conditions necessary for the transduction of local pH changes into electrical signals. PMID- 15115706 TI - Coordinate downregulation of CaM kinase II and phospholamban accompanies contractile phenotype transition in the hyperthyroid rabbit soleus. AB - This study investigated the effects of l-thyroxine-induced hyperthyroidism on Ca(2+)/calmodulin (CaM)-dependent protein kinase (CaM kinase II)-mediated sarcoplasmic reticulum (SR) protein phosphorylation, SR Ca(2+) pump (Ca(2+) ATPase) activity, and contraction duration in slow-twitch soleus muscle of the rabbit. Phosphorylation of Ca(2+)-ATPase and phospholamban (PLN) by endogenous CaM kinase II was found to be significantly lower (30-50%) in soleus of the hyperthyroid compared with euthyroid rabbit. Western blotting analysis revealed higher levels of sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA) 1 ( approximately 150%) Ca(2+) pump isoform, unaltered levels of SERCA2 Ca(2+) pump isoform, and lower levels of PLN ( approximately 50%) and delta-, beta-, and gamma-CaM kinase II (40 approximately 70%) in soleus of the hyperthyroid rabbit. SR vesicles from hyperthyroid rabbit soleus displayed approximately twofold higher ATP-energized Ca(2+) uptake and Ca(2+)-stimulated ATPase activities compared with that from euthyroid control. The V(max) of Ca(2+) uptake (in nmol Ca(2+).mg SR protein(-1).min(-1): euthyroid, 818 +/- 73; hyperthyroid, 1,649 +/- 90) but not the apparent affinity of the Ca(2+)-ATPase for Ca(2+) (euthyroid, 0.97 +/- 0.02 microM, hyperthyroid, 1.09 +/- 0.04 microM) differed significantly between the two groups. CaM kinase II-mediated stimulation of Ca(2+) uptake by soleus muscle SR was approximately 60% lower in the hyperthyroid compared with euthyroid. Isometric twitch force of soleus measured in situ was significantly greater ( approximately 36%), and the time to peak force and relaxation time were significantly lower ( approximately 30-40%), in the hyperthyroid. These results demonstrate that thyroid hormone-induced transition in contractile properties of the rabbit soleus is associated with coordinate downregulation of the expression and function of PLN and CaM kinase II and selective upregulation of the expression and function of SERCA1, but not SERCA2, isoform of the SR Ca(2+) pump. PMID- 15115707 TI - Selective regulation by delta-PKC and PI 3-kinase in the assembly of the antiapoptotic TNFR-1 signaling complex in neutrophils. AB - TNF is implicated in the attenuation of neutrophil constitutive apoptosis during sepsis. Antiapoptotic signaling is mediated principally through the TNF receptor 1 (TNFR-1). In adherent neutrophils, when beta-integrin signaling is activated, TNF phosphorylates TNFR-1 and activates prosurvival and antiapoptotic signaling. Previously, we identified the delta-PKC isotype and phosphatidylinositol (PI) 3 kinase as critical regulators of TNF signaling in adherent neutrophils. Both kinases associate with TNFR-1 in response to TNF and are required for TNFR-1 serine phosphorylation, NF-kappaB activation, and inhibition of apoptosis. The purpose of this study was to examine the role of delta-PKC and PI 3-kinase in the assembly of TNFR-1 signaling complex that regulates NF-kappaB activation and antiapoptotic signaling. Coimmunoprecipitation studies established that PI 3 kinase, delta-PKC, and TNFR-1 formed a signal complex in response to TNF. delta PKC recruitment required both delta-PKC and PI 3-kinase activity, whereas PI 3 kinase recruitment was delta-PKC independent, suggesting that PI 3-kinase acts upstream of delta-PKC. An important regulatory step in control of antiapoptotic signaling is the assembly of the TNFR-1-TNFR-1-associated death domain protein (TRADD)-TNFR-associated factor 2 (TRAF2)-receptor interacting protein (RIP) complex that controls NF-kappaB activation. Inhibition of either delta-PKC or PI 3-kinase decreased TNF-mediated recruitment of RIP and TRAF2 to TNFR-1. In contrast, TRADD recruitment was enhanced. Thus delta-PKC and PI 3-kinase are positive regulators of TNF-mediated association of TRAF2 and RIP with TNFR-1. Conversely, these kinases are negative regulators of TRADD association. These results suggest that delta-PKC and PI 3-kinase regulate TNF antiapoptotic signaling at the level of the TNFR-1 through control of assembly of a TNFR-1 TRADD-RIP-TRAF2 complex. PMID- 15115708 TI - Ciprofloxacin-induced toxic epidermal necrolysis (TEN) in a nonagerian: a case report. AB - A 93-year-old lady developed extensive skin lesions following treatment with ciprofloxacin for urinary tract infection. Skin biopsy confirmed toxic epidermal necrolysis. She died within 48 hours of onset of her skin signs in spite of active management. Toxic epidermal necrolysis is a rare but serious side-effect of ciprofloxacin. PMID- 15115709 TI - The German etanercept registry for treatment of juvenile idiopathic arthritis. AB - OBJECTIVE: To describe a registry set up to monitor children treated with etanercept in Germany and Austria. METHODS: Giannini's criteria, duration of morning stiffness, number of swollen, tender and contracted joints, adverse events, and reasons for discontinuation were assessed. RESULTS: 322 patients with juvenile idiopathic arthritis (JIA) and 12 additional patients with non-JIA rheumatic diagnoses were included. Therapeutic efficacy was observed from one month after treatment was started. The number of patients with significant improvement and the degree of improvement increased during the first year. The mean (SD) number of tender and swollen joints decreased from 9 (9) and 8.4 (9) to 3.0 (6.5) and 4.5 (7) after one month, and to 2.2 (5.5) and 3.3 (5.5) after three months; morning stiffness decreased from 45 (65) minutes to 12 (30) and 7 (19) after one and three months (p<0.001 for all). Using Gianinni's criteria of 30%, 50%, and 70% improvement, a therapeutic response in JIA patients was achieved in, respectively, 66%, 54%, and 30% after one month, 78%, 61%, and 38% after three months, and 83%, 72%, and 52% after six months. Therapeutic efficacy was lower in patients with systemic onset arthritis. Overall tolerability was good: in 592 patient treatment-years there were 69 reports of adverse events in 56 patients, including one CNS demyelination. There were no opportunistic infections or lupus like reactions. Treatment was discontinued in 53 JIA patients, in 25 because of lack of efficacy. CONCLUSION: Etanercept treatment was safe and led to a significant improvement in most JIA patients resistant to conventional treatment. PMID- 15115710 TI - Efficacy of etanercept in the treatment of acute, severe sciatica: a pilot study. AB - OBJECTIVES: To explore the efficacy of a tumour necrosis factor alpha (TNFalpha) inhibitor (etanercept, Enbrel) in patients with severe sciatica. METHODS: A pilot study of etanercept was conducted in patients admitted to hospital for acute severe sciatica. Ten consecutive patients received three subcutaneous injections of etanercept (25 mg every 3 days) in addition to standard analgesia. Response was evaluated at day 10 (T1) and week 6 (T2) using a visual analogue scale for leg pain (VASL) and for low back pain (VASB), and two validated functional scores: the Oswestry disability index (ODI) and the Roland Morris disability questionnaire (RMDQ). The control group consisted of 10 patients with severe sciatica, who took part in an observational study on i.v. methylprednisolone. RESULTS: In the etanercept group all variables improved: VASB from 36 to 7; VASL from 74 to 12; RMDQ from 17.8 to 5.8, and ODI from 75.4 to 17.3; all p<0.001. Pain (VASL and VASB: p<0.001) and ODI (p<0.05) were significantly better in the etanercept group than in the methylprednisolone group. CONCLUSION: In this open, historical group controlled study, patients with severe sciatica had sustained improvement after a short treatment with etanercept that was better than standard care plus a short course of methylprednisolone. These results suggest that inhibition of TNFalpha is beneficial in the treatment of sciatica and support a pathological role for TNFalpha in the pathogenesis of sciatica. These results need to be confirmed by a randomised controlled trial. PMID- 15115711 TI - Contribution of polymorphisms in the apolipoprotein AI-CIII-AIV cluster to hyperlipidaemia in patients with gout. AB - BACKGROUND: Studies have shown that hyperuricaemia is independently related to the insulin resistance syndrome and that polymorphisms of the apolipoprotein AI CIII-AIV cluster are also related to insulin resistance. OBJECTIVE: To study the prevalence of polymorphisms of the apolipoprotein AI-CIII-AIV cluster in persons with gout and to determine whether these polymorphisms contribute to the pathophysiology of gout or to altered lipid concentrations. METHODS: Plasma cholesterol, triglycerides, uric acid, VLDL, LDL, IDL, and HDL triglycerides, cholesterol, and the renal excretion of uric acid were measured in 68 patients with gout with gout and 165 healthy subjects. Polymorphisms were studied by amplification and RFLP in all subjects, using XmnI and MspI in the apolipoprotein AI gene and SstI in the apolipoprotein CIII gene. RESULTS: The A allele at position -75 bp in the apolipoprotein AI gene was more common in patients with gout than in controls (p = 0.01). Levels of cholesterol, triglycerides, uric acid, basal glycaemia, and HDL cholesterol were higher in the patients (p<0.001). In the patients there was also an interaction between mutations at the two polymorphic loci studied in the apolipoprotein AI gene (p = 0.04). An absence of the mutation at position -75 bp of the apolipoprotein AI gene resulted in increased plasma triglyceride levels. CONCLUSIONS: Gouty patients have an altered allelic distribution in the apolipoprotein AI-CIII-AIV cluster, which could lead to changes in levels of lipoproteins. This is not caused by a single mutation but rather by a combination of different mutations. PMID- 15115712 TI - Antinuclear and antiphospholipid autoantibodies in patients with peripheral arterial occlusive disease. PMID- 15115713 TI - Differential effects of leflunomide and methotrexate on cytokine production in rheumatoid arthritis. AB - BACKGROUND: T cells have a pivotal role in RA. Leflunomide inhibits pyrimidine biosynthesis, to which T cells are especially susceptible, and therefore may have a different cytokine profile than methotrexate. MATERIALS AND METHODS: Serum samples of 100 patients with RA, treated with leflunomide (n = 50) or methotrexate (n = 50), were collected at baseline, after 16 weeks and after 1 year's treatment. Serum levels of interleukin 6 (IL6), and interferon (IFN) gamma were determined by ELISA. Additionally, peripheral blood mononuclear cells (PBMC) of five healthy volunteers and three patients with RA were isolated and the effects of the active metabolite of leflunomide (A77-1726, 0-200 mmol/l) on cell proliferation and on IL6 and IFNgamma production were determined by ELISA. In peripheral blood lymphocytes (PBL) and monocytes (PBM) from two healthy volunteers the effects of A77-1726 on IL6 production were measured by ELISA and PCR. RESULTS: Mean (SEM) serum levels of IFNgamma were significantly reduced after leflunomide treatment (baseline 43 (10) pg/ml; 1 year 29 (7) (p = 0.015), but there was no change in IL6 levels (baseline 158 (41), 1 year 151 (48)). Both IFNgamma and IL6 levels were significantly reduced after methotrexate treatment. This observation was supported by in vitro experiments. The production of IFNgamma by PBL was inhibited by A77-1726, but IL6 production by PBM was not inhibited. CONCLUSION: The differential effect on IFNgamma and IL6 production supports the hypothesis that activated T cells are preferentially inhibited by leflunomide. An explanation may be either inhibition of uridine synthesis or effects on signal transduction pathways. PMID- 15115714 TI - Rate of cartilage loss at two years predicts subsequent total knee arthroplasty: a prospective study. AB - OBJECTIVE: To determine whether cartilage volume loss is an independent predictor of knee replacement. DESIGN: Prospective community based, four year prospective cohort study. METHODS: 123 subjects with mild to moderate symptomatic radiographic knee osteoarthritis were recruited by either advertising, the Victorian branch of the Arthritis Foundation of Australia, treating general practitioners, orthopaedic surgeons, or rheumatologists; 113 completed the study. Magnetic resonance imaging was carried out at baseline and at 2 years on the symptomatic knee. Rate of change in tibial cartilage volume was calculated. Subjects were then followed up at year 4 to determine whether they had undergone a knee replacement. RESULTS: The rate of tibial cartilage loss over two years was an independent predictor of knee replacement at four years. For every 1% increase in the rate of tibial cartilage loss there was a 20% increase risk of undergoing a knee replacement at four years (95% confidence interval, 10% to 30%). Those in the highest tertile of tibial cartilage loss had 7.1 (1.4 to 36.5) higher odds of undergoing a knee replacement than those in the lowest tertile. WOMAC score at baseline, female sex, and tibial bone size (but not age and radiographic score) were also predictors of knee replacement. CONCLUSIONS: The data suggest that treatment targeted at reducing the rate of knee cartilage loss in subjects with symptomatic osteoarthritis may delay knee replacement. This has important implications in terms of prevention and therapeutic interventions in osteoarthritis. PMID- 15115715 TI - MyD88 is required for mounting a robust host immune response to Streptococcus pneumoniae in the CNS. AB - Myeloid differentiation factor 88 (MyD88) is an essential intracellular signal transducer in Toll-like receptor (TLR) and interleukin (IL)-1 receptor family member-mediated cell activation. In order to characterize the role of MyD88 in pneumococcal meningitis we used gene-targeted mice lacking functional MyD88 expression. At 24 h after intracisternal infection, MyD88- deficient mice displayed a markedly diminished inflammatory host response in the CNS, as evidenced by reduced CSF pleocytosis and expression of cytokines, chemokines and complement factors. The reduced CNS inflammation was paralleled by a marked reduction in the prognostic relevant CNS complications, such as brain oedema formation. Nevertheless, MyD88 deficiency was associated with a worsening of disease which seemed to be attributable to severe bacteraemia. This notion was supported by the unexpected observation that infected MyD88-deficient mice displayed enhanced mRNA expression of inflammatory mediators [such as the proinflammatory cytokine tumour necrosis factor alpha (TNF-alpha) and the CXC chemokine macrophage inflammatory protein (MIP-2)] in the lung and consequently increased cell influx in the bronchoalveolar lavage fluid, compared with infected wild-type mice. Thus, the present study demonstrated for the first time an important role of MyD88 in immune activation to bacterial pathogens within the CNS. The role played by MyD88 in mounting an immune response to Streptococcus pneumoniae, however, seems to be dependent on the anatomical compartment involved. PMID- 15115716 TI - Gonadotropin surge-induced differential upregulation of collagenase-1 (MMP-1) and collagenase-3 (MMP-13) mRNA and protein in bovine preovulatory follicles. AB - The ovulatory process is characterized by focalized extracellular matrix degradation at the apex of preovulatory follicles. Many studies have implicated the matrix metalloproteinases (MMPs) as potential mediators of follicle rupture. Objectives of this study were to determine localization and effect of the gonadotropin surge on temporal expression of MMP-1 and MMP-13 in bovine preovulatory follicles. Samples were collected at 0, 6, 12, 18, 24, and 48 h (corpora lutea) after GnRH injection (n = 5-6 per time point) and amounts of MMP 1 and MMP-13 mRNA and protein determined using dot blot or semiquantitative RT PCR and Western blot analyses. Samples were also collected at 0 and 20 h after GnRH injection for immunohistochemical localization of MMP-1 and MMP-13. Results indicate that follicular expression of MMP-1 and MMP-13 increased following the gonadotropin surge. Abundance of MMP-1 mRNA increased at 6, 12, and 48 h post GnRH injection. Immunoreactive MMP-1 was localized to granulosal and thecal layers of preovulatory follicles. Amounts of MMP-1 protein increased in both the apex and the base of preovulatory follicles. Abundance of MMP-13 mRNA increased at 6, 24, and 48 h post GnRH injection. Amounts of MMP-13 protein also increased in the follicular apex and base. Immunoreactive MMP-13 was localized to granulosal and thecal layers of preovulatory follicles. Results indicate MMP-1 and MMP-13 are increased in bovine preovulatory follicles following the gonadotropin surge but do not support a requirement for differential up regulation of MMP-1 and MMP-13 (follicular apex vs. base) for the preovulatory collagenolysis required for follicle rupture. PMID- 15115717 TI - Vitellogenesis in the sand shrimp, metapenaeus ensis: the contribution from the hepatopancreas-specific vitellogenin gene (MeVg2). AB - An additional vitellogenin gene (MeVg2) that is structurally different from MeVg1 was cloned and characterized from the shrimp Metapenaeus ensis. The MeVg2 gene consists of fewer exons-introns and is most likely evolved from the MeVg1 gene. The cDNA for MeVg2 is 8.0 kilobases (kb) in size, and the deduced MeVg2 precursor shared an overall 54% amino sequence identity to the MeVg1 gene of the same shrimp. As compared to the MeVg1 precursor, MeVg2 precursor consists of more potential subunit cleavage sites, suggesting that the precursor may be processed into many smaller subunits. The MeVg2 is expressed only in the hepatopancreas, and the expression level of MeVg2 in adult female increases from the early vitellogenic stage, reaching a maximum at the middle vitellogenic stage, and remains high toward the end of vitellogenic cycle. In addition to the 8-kb mRNA, smaller transcripts of 1.5-2.5 kb for MeVg2 were identified, and the 8-kb transcript only constitutes less than 10% of the overall MeVg2-derived transcripts. To confirm the presence of the small transcripts, we screened the shrimp hepatopancreas cDNA library and isolated two smaller MeVg2-specific cDNA clones. These clones shared greater than 99% overall identity to the corresponding C-terminal region of the MeVg2 precursor, suggesting that an alternative expression/ splicing of the MeVg2 gene occurred. By immunohistochemical analysis, vitellin-immunopositive signals were localized in the lumen and extracellular fraction of the hepatopancreas. Amino acid sequence determination of the tissue protein and secreted protein from the hepatopancreas revealed that the 76-kDa vitellogenin subunit is most likely processed into smaller-sized subunits. Taken together, these results suggest that the hepatopancreas is an important organ for the synthesis of vitellogenin and may contribute to vitellogenesis by producing a large quantity of smaller MeVg2 subunit for ovarian uptake. PMID- 15115718 TI - Culture conditions and single growth factors affect fate determination of mouse spermatogonial stem cells. AB - Cell fate determination between self-renewal or differentiation of spermatogonial stem cells (SSCs) in the testis is precisely regulated to maintain normal spermatogenesis. However, the mechanisms underlying the process remain elusive. To address the problem, we developed a model SSC culture system, first, by establishing techniques to obtain enriched populations of stem cells, and second, by establishing a serum-free culture medium. Flow cytometric cell sorting and the SSC transplantation assay demonstrated that Thy-1 is a unique surface marker of SSCs in neonatal, pup, and adult testes of the mouse. Although the surface phenotype of SSCs is major histocompatibility complex class I(-) Thy-1(+) alpha 6 integrin(+) alpha v-integrin(-/dim) throughout postnatal life, the most enriched population of SSCs was obtained from cryptorchid adult testes by cell-sorting techniques based on Thy-1 expression. This enriched population of SSCs was used to develop a culture system that consisted of serum-free defined medium and STO (SIM mouse embryo-derived thioguanine and ouabain resistant) feeders, which routinely maintained stem cell activity for 1 wk. Combining the culture system and the transplantation assay provided a mechanism to study the effect of single growth factors. A negative effect was demonstrated for several concentrations of basic fibroblast growth factor and leukemia inhibitory factor, whereas glial cell line-derived neurotrophic factor and stem cell factor appeared to have a positive effect on stem cell maintenance. The stem cell enrichment strategies and the culture methods described provide a reproducible and powerful assay system to establish the effect of various environmental factors on SSC survival and replication in vitro. PMID- 15115719 TI - Biosynthesis of the canine zona pellucida requires the integrated participation of both oocytes and granulosa cells. AB - In the dog, attempts to localize the expression of zona pellucida (ZP) proteins during folliculogenesis have failed to demonstrate conclusively whether any or all of the zona proteins are synthesized in the oocyte or the granulosa cells. Probing of paraformaldehyde-fixed prepubertal canine ovarian tissue sections with a panel of fluorescently conjugated lectins localized the expression of glycoproteins during folliculogenesis. We confirm that six lectins (PSA, s-WGA, ECL, GSL-II, LEL, and STL) consistently labeled the ZP and adjacent granulosa cells of the developing follicle and that canine ZP expresses beta gal(1,4)glcNAc, beta-gal(1,3)galNac, alpha-mannose, and terminal sialic acid residues in a developmentally specific manner. Riboprobes for canine ZPA and ZPC genes were produced and used for in situ hybridization studies of mRNA expression in canine folliculogenesis. In addition, we isolated a partial cDNA transcript from total ovarian RNA for the canine ZPB gene having a high degree of sequence identity with the felid and porcine ZPB homologues. Subsequently, the ZPA gene transcripts were localized to the cytoplasm of oocytes in primordial, primary, and early secondary follicles. We then localized expression of ZPB and ZPC gene transcripts to the granulosa cells of growing follicles, but not in squamous granulosa cells of primordial follicles or oocytes. These observations indicate that in the juvenile canine ovary, the oocyte is responsible for synthesis of the ZPA protein and directing synthesis of the ZPB and ZPC proteins by the granulosa cells and that ZP gene transcription occurs in a sequential manner during folliculogenesis. PMID- 15115720 TI - Interaction of sperm with purified native chicken ZP1 and ZPC proteins. AB - The avian perivitelline membrane (PVM) is the site of initial contact between sperm and egg. It consists of only two major components, which are both homologues of the mammalian zona pellucida (ZP) proteins, and belong to the ZP1 and ZPC families, respectively. We have established a method to isolate large quantities of both native avian ZP proteins and have used these preparations to investigate their sperm-binding capacities. Chicken ZPC forms multimeric structures of defined size and binds to an approximately 180-kDa protein complex present in rooster sperm extracts. Based on experiments using both PVM and isolated proteins, we show that chicken ZP1 is proteolytically degraded by a sperm-associated protease but that chicken ZPC remains intact. An antiserum directed against chicken ZP1 is capable of inhibiting sperm binding to the PVM. Taken together, these data suggest that ZP1, in addition to ZPC, plays a major role in the initial interactions between sperm and egg. PMID- 15115721 TI - Cloning of epidermal growth factor (EGF) and EGF receptor from the zebrafish ovary: evidence for EGF as a potential paracrine factor from the oocyte to regulate activin/follistatin system in the follicle cells. AB - In the present study, we cloned full-length cDNAs for epidermal growth factor (EGF), EGF receptor (EGFR), and three truncated forms of EGFR (EGFR15, 12, and 8) from the zebrafish ovary. Zebrafish EGF was predominantly expressed in the ovary and testis, while EGFR and its truncated forms were highly expressed in all tissues examined except the liver. In the ovary, the expression of EGF seemed to be more abundant in the follicles of early stages, while EGFR had much higher expression levels at later stages. Interestingly, although EGF was expressed in both the follicle cells and oocytes, its expression level was significantly higher in the oocytes. However, the expression of EGFR was mainly restricted to the follicle cells with little expression in the oocytes. The unique spatial patterns of EGF and EGFR expression within the follicle suggest that EGF may serve as a messenger from the oocyte to signal the follicle cells. EGF strongly stimulated the expression of both activin beta A and beta B, while it suppressed basal and hCG-induced follistatin expression in cultured follicle cells. These results, together with the evidence that EGF was predominantly expressed in the oocytes whereas EGFR was expressed in the follicle cells, strongly suggest that EGF is likely a potential paracrine/juxtacrine factor from the oocytes to regulate the function of the follicle cells. PMID- 15115722 TI - Intra-oocyte localization of MAD2 and its relationship with kinetochores, microtubules, and chromosomes in rat oocytes during meiosis. AB - The present study was designed to investigate subcellular localization of MAD2 in rat oocytes during meiotic maturation and its relationship with kinetochores, chromosomes, and microtubules. Oocytes at germinal vesicle (GV), prometaphase I (ProM-I), metaphase I (M-I), anaphase I (A-I), telophase I (T-I), and metaphase II (M-II) were fixed and immunostained for MAD2, kinetochores, microtubules and chromosomes. The stained oocytes were examined by confocal microscopy. Some oocytes from GV to M-II stages were treated by a microtubule disassembly drug, nocodazole, or treated by a microtubule stabilizer, Taxol, before examination. Anti-MAD2 antibody was also injected into the oocytes at GV stage and the injected oocytes were cultured for 6 h for examination of chromosome alignment and spindle formation. It was found that MAD2 was at the kinetochores in the oocytes at GV and ProM-I stages. Once the oocytes reached M-I stage in which an intact spindle was formed and all chromosomes were aligned at the equator of the spindle, MAD2 disappeared. However, when oocytes from GV to M-II stages were treated by nocodazole, spindles were destroyed and MAD2 was observed in all treated oocytes. When nocodazole-treated oocytes at M-I and M-II stages were washed and cultured for spindle recovery, it was found that, once the relationship between microtubules and chromosomes was established, MAD2 disappeared in the oocytes even though some chromosomes were not aligned at the equator of the spindle. On the other hand, when oocytes were treated with Taxol, MAD2 localization was not changed and was the same as that in the control. However, immunoblotting of MAD2 indicated that MAD2 was present in the oocytes at all stages; nocodazole and Taxol treatment did not influence the quantity of MAD2 in the cytoplasm. Significantly higher proportions of anti-MAD2 antibody-injected oocytes proceeded to premature A-I stage and more oocytes had misaligned chromosomes in the spindles. The present study indicates that MAD2 is a spindle checkpoint protein in rat oocytes during meiosis. When the spindle was destroyed by nocodazole, MAD2 was reactivated in the oocytes to overlook the attachment between chromosomes and microtubules. However, in this case, MAD2 could not check unaligned chromosomes in the recovered spindles, suggesting that a normal chromosome alignment is maintained only in the oocytes without any microtubule damages during maturation. PMID- 15115723 TI - Extracellular matrix: recent advances on its role in junction dynamics in the seminiferous epithelium during spermatogenesis. AB - Spermatogenesis takes place in the seminiferous epithelium of the mammalian testis in which one type A1 spermatogonium (diploid, 2n) gives rise to 256 spermatids (haploid, 1n). To accomplish this, developing germ cells, such as preleptotene and leptotene spermatocytes, residing in the basal compartment of the seminiferous epithelium must traverse the blood-testis barrier (BTB) entering into the adluminal compartment for further development into round, elongating, and elongate spermatids. Recent studies have shown that the basement membrane in the testis (a modified form of extracellular matrix, ECM) is important to the event of germ cell movement across the BTB because proteins in the ECM were shown to regulate BTB dynamics via the interactions between collagens, proteases, and protease inhibitors, possibly under the regulation of cytokines. While these findings are intriguing, they are not entirely unexpected. For one, the basement membrane in the testis is intimately associated with the BTB, which represents the basolateral region of Sertoli cells. Also, Sertoli cell tight junctions (TJs) that constitute the BTB are present side-by-side with cell-cell actin-based adherens junctions (AJ, such as basal ectoplasmic specialization [ES]) and intermediate filament-based desmosome-like junctions. As such, the relative morphological layout between TJs, AJs, and desmosome-like junctions in the seminiferous epithelium is in sharp contrast to other epithelia where TJs are located at the apical portion of an epithelium or endothelium, furthest away from ECM, to be followed by AJs and desmosomes, which in turn constitute the junctional complex. For another, anchoring junctions between a cell epithelium and ECM found in multiple tissues, also known as focal contacts (or focal adhesion complex, FAC, an actin-based cell-matrix anchoring junction type), are the most efficient junction type that permits rapid junction restructuring to accommodate cell movement. It is therefore physiologically plausible, and perhaps essential, that the testis is using some components of the focal contacts to regulate rapid restructuring of AJs between Sertoli and germ cells when germ cells traverse the seminiferous epithelium. Indeed, recent findings have shown that the apical ES, a testis-specific AJ type in the seminiferous epithelium, is equipped with proteins of FAC to regulate its restructuring. In this review, we provide a timely update on this exciting yet rapidly developing field regarding how the homeostasis of basement membrane in the tunica propria regulates BTB dynamics and spermatogenesis in the testis, as well as a critical review on the molecular architecture and the regulation of ES in the seminiferous epithelium. PMID- 15115724 TI - Regulation of ubiquitin-proteasome pathway on pig oocyte meiotic maturation and fertilization. AB - Degradation of proteins mediated by the ubiquitin-proteasome pathway (UPP) plays essential roles in the eukaryotic cell cycle. The main aim of the present study was to analyze the functional roles and regulatory mechanisms of the UPP in pig oocyte meiotic maturation, activation, and early embryo mitosis by drug treatment, Western blot analysis, and confocal microscopy. By using the hypoxanthine-maintained meiotic arrest model, we showed that the meiotic resumption of both cumulus-enclosed oocytes and denuded oocytes was stimulated in a dose- and time-dependent manner by two potent and cell-permeable proteasome inhibitors. Both the mitogen-activated protein kinase (MAPK) kinase inhibitor U0126 and the maturation-promoting factor inhibitor roscovitine overcame the stimulation of germinal vesicle breakdown induced by proteasome inhibitors. The phosphorylation of MAPK and p90rsk and the expression of cyclin B1 increased in a dose- and time-dependent manner when treated with proteasome inhibitors during oocyte in vitro-maturation culture. Both U0126 and roscovitine inhibited the phosphorylation of MAPK and p90rsk, and the synthesis of cyclin B1 stimulated by proteasome inhibitors. When matured oocytes were pretreated with proteasome inhibitors and then fertilized or artificially activated, the second polar body emission and the pronuclear formation were inhibited, and the dephosphorylation of MAPK and p90rsk as well as the degradation of cyclin B1 that should occur after oocyte activation were also inhibited. We also investigated, to our knowledge for the first time, the subcellular localization of 20S proteasome alpha subunits at different stages of oocyte and early embryo development. The 20S proteasome alpha subunits were accumulated in the germinal vesicle, around the condensed chromosomes at prometaphase, with spindle at metaphase I and II, the region between the separating chromosomes, and especially the midbody at anaphase I and telophase I, the pronucleus, and the nucleus in early embryonic cells. In conclusion, our results suggest that the UPP is important at multiple steps of pig oocyte meiosis, fertilization, and early embryonic mitosis and that it may play its roles by regulating cyclin B1 degradation and MAPK/p90rsk phosphorylation. PMID- 15115725 TI - Reproductive physiology and development of artificial insemination technology in killer whales (Orcinus orca). AB - Research was conducted to define the basic reproductive physiology of killer whales (Orcinus orca) and to use this knowledge to facilitate the development of artificial insemination procedures. The specific objectives were 1) to determine the excretory dynamics of urinary LH and ovarian steroid metabolites during the estrous cycle; 2) to evaluate the effect of an exogenously administered, synthetic progesterone analog on reproductive hormone excretion; 3) to validate the use of transabdominal ultrasound for ovarian evaluation and timing of ovulation; 4) to examine the quality of semen after liquid storage and cryopreservation; and 5) to develop an intrauterine insemination technique. Based on urinary endocrine monitoring of 41 follicular phases and 26 complete cycles from five females, estrous cycles were 41 days long and comprised a 17-day follicular phase and a 21-day luteal phase. A consistent temporal relationship was observed between peak estrogen conjugates and the LH surge, the latter of which occurred approximately 0.5 days later. Two animals placed on oral altrenogest (three separate occasions for 30, 17, and 31 days, respectively) excreted peak urinary estrogen concentrations 25 days after withdrawal that were followed by sustained elevations in urinary pregnanediol-3alpha-glucuronide excretion. Mean preovulatory follicle diameter was 3.9 cm (n = 6), and ovulation occurred 38 h (n = 5) after the peak of the LH surge. Based on visual estimates of motility, liquid-stored semen maintained 92% of its raw ejaculate sperm motility index (total progressive motility x kinetic rating [0-5 scale, where 0 = no movement and 5 = rapid progressive movement]) when held at 4 degrees C for 3 days postcollection. Semen cryopreserved using a medium freezing rate demonstrated good postthaw total motility (50%), progressive motility (94%), and kinetic rating (3.5). Insemination during eight estrous cycles resulted in three pregnancies (38%), two from liquid-stored and one from cryopreserved semen. Two calves were delivered after gestation lengths of 552 and 554 days, respectively. These data demonstrate the potential of noninvasive endocrine monitoring combined with serial ultrasonography to improve our understanding of the reproductive biology of cetaceans. This fundamental knowledge was essential for ensuring the first successful conceptions, resulting in live offspring, using artificial insemination in any cetacean species. PMID- 15115726 TI - Regulation of N-methyl-D-aspartate receptor subunit expression in the fetal guinea pig brain. AB - N-methyl-d-aspartate receptors (NMDARs) are critical for neuronal maturation and synaptic formation as well as for the onset of long-term potentiation, a process critical to learning and memory in postnatal life. In the current study, we demonstrated that NMDAR subunits undergo spatial, temporal, and sex-specific regulation. During development, we observed increasing NR1 and NR2A expression at the same time as levels of NR2B subunits decreased in the hippocampus and cortex in the fetal guinea pig. We have also shown that glucocorticoids can modulate fetal NMDAR subunit expression in a sex-specific fashion. This is clinically important because synthetic glucocorticoids are administered to pregnant women at risk of preterm labor. Repeated exposure to exogenous glucocorticoids caused a dose-dependent decrease in NR1 mRNA levels and increased NR2A mRNA expression in the female hippocampus at Gestational Day 62. There are significant changes in NMDAR subunit expression in late gestation. It is possible that these alter NMDA dependent signaling at this time. Prenatal exposure to exogenous glucocorticoids modifies the trajectory of NMDAR subunit expression in females but not in males. PMID- 15115727 TI - Reduced growth hormone secretion prolongs puberty but does not delay the developmental increase in luteinizing hormone in the absence of gonadal negative feedback. AB - Previous studies have shown that the growth hormone (GH) axis is important for timing the later stages of puberty in female monkeys. However, it is not clear whether these growth-related signals are important for the initiation of puberty and early pubertal events. The present study, using female rhesus monkeys, used two approaches to answer this question. Experiment 1 tested the hypothesis that reduced GH secretion would blunt the rise in nocturnal LH secretion in young (17 mo; n = 7) but not older adolescent ovariectomized females (29 mo; n = 6). Reduced GH secretion was induced by treating females with the sustained release somatostatin analogue formulation, Sandostatin LAR (625 microg/kg). Morning (0900 0930 h) and evening (2200-2230 h) concentrations of bioactive LH were higher in older adolescent compared to young adolescent females. However, diurnal concentrations were not affected by the inhibition of GH secretion in either age group when compared to the placebo-treated, control condition. Experiment 2 tested the hypothesis that reduced GH secretion induced in young juvenile females would delay the initial increase in nocturnal LH secretion and subsequent early signs of puberty. In order to examine this hypothesis, puberty in control females (n = 7) was compared to those in which puberty had been experimentally arrested until a late adolescent age (29 mo) by the use of a depot GnRH analogue, Lupron (750 microg kg(-1) mo(-1); n = 7). Once the analogue treatment was discontinued, the progression of puberty was compared to a group treated in a similar fashion but made GH deficient by continuous treatment with Sandostatin LAR (n = 6). Puberty occurred as expected in control females with the initial rise in evening LH at 21 mo, menarche at 22 mo, and first ovulation at 30 mo. As expected, Lupron arrested reproductive maturation, but elevations in morning and evening LH and menarche occurred within 2 mo of the cessation of Lupron in both Lupron and Lupron-GH-suppressed females. In contrast, first ovulation was delayed significantly in the Lupron-GH-suppressed females (41 mo) compared to the Lupron only females (36 mo). These data indicate that within this experimental model, reduced GH secretion does not perturb the early stages of puberty but supports previous observations that the GH axis is important for timing the later stages of puberty and attainment of fertility. Taken together, the data indicate that factors that reduce GH secretion may have a deleterious effect on the completion of puberty. PMID- 15115728 TI - Gamma-H2AX expression pattern in non-irradiated neonatal mouse germ cells and after low-dose gamma-radiation: relationships between chromatid breaks and DNA double-strand breaks. AB - The DNA double-strand breaks (DSBs) are considered to be the most relevant lesions for the deleterious effects of ionizing radiation exposure. The discovery that the induction of DSBs is rapidly followed by the phosphorylation of H2AX histone at Ser-139, favoring repair protein recruitment or access, opens the possibility for a wide range of research. This phosphorylated histone, named gamma-H2AX, has been shown to form foci in interphase nuclei as well as megabase chromatin domains surrounding the DNA lesion on chromosomes. Using detection of gamma-H2AX on germ cell mitotic chromosomes 2 h after gamma-irradiation, we studied radiation-induced DSBs during the G(2)/M phase of the cell cycle. We show that 1) non-irradiated neonatal germ cells express gamma-H2AX with variable patterns at metaphase, 2) gamma-irradiation induces foci whose number increases in a dose-dependent manner, 3) some foci correspond to visible chromatid breaks or exchanges, 4) sticky chromosomes characterizing cell radiation exposure during mitosis are a consequence of DSBs, and 5) gamma-H2AX remains localized at the sites of the lesions even after end-joining has taken place. This suggests that completion of DSB repair does not necessarily imply disappearance of gamma-H2AX. PMID- 15115729 TI - In vivo and in vitro regulation of Akt activation in human endometrial cells is estrogen dependent. AB - Estrogen-bound estrogen receptors (ER) alpha and beta classically activate gene expression after binding to the estrogen response element in the promoter regions of target genes. Estrogen also has rapid, nongenomic effects. It activates several membranous or cytoplasmic kinase cascades, including the phosphatidylinositol 3-phosphate (PI3K/Akt) cascade, a signaling pathway that plays a key role in cell survival and apoptosis. Normal human endometrium is exposed to variable levels of steroid hormones throughout the menstrual cycle. We hypothesized that Akt phosphorylation in human endometrium may vary with the menstrual cycle and in early pregnancy and that fluctuations in estrogen level may play a role in Akt activation in endometrial cells. We analyzed Akt phosphorylation using in vivo and in vitro techniques, including Western blot, immunohistochemistry, and immunocytochemistry. Estradiol significantly increased Akt phosphorylation in endometrial cells. Rapid stimulation of Akt activation in cultured stromal cells was observed. Akt phosphorylation by estradiol was inhibited by the PI3K inhibitor, wortmannin, but not by the ER antagonist, ICI 182 780. The maximal effect on Akt activity was observed following 5-15 min of estradiol treatment. Our results suggest that estradiol may directly affect PI3K related signaling pathway by increasing the phosphorylation of Akt in endometrial cells. Thus, estradiol may exert part of its proliferative and antiapoptotic effects by a nongenomic manner through the Akt signaling pathway. PMID- 15115730 TI - Activation of p38 MAPK during porcine oocyte maturation. AB - The p38 MAPK is a member of the mitogen-activated protein kinase (MAPK) family that participates in a signaling cascade in response to cytokines and stress in somatic cells. The present study was designed to investigate the expression and possible function of p38 MAPK in porcine oocytes during maturation. In immunoblots, p38 MAPK was detected in oocytes and cumulus cells. Its activity was determined during oocyte maturation in vitro by the phosphorylation of its substrate, activated transcription factor 2. As ERK1/2, oocyte p38 MAPK became active around germinal vesicle breakdown (GVBD) and maintained activity until metaphase II (MII). Immunofluorescent microscopy showed phosphorylated p38 MAPK accumulated in the nucleus before GVBD and localized in the cytoplasm and around chromosomes from metaphase I (MI) to MII. In cultured cumulus-oocyte complexes, a specific inhibitor of p38 MAPK, SB203580, inhibited phosphorylation of p38 MAPK in cumulus cells and blocked both FSH-induced cumulus expansion and meiotic resumption of oocytes. During spontaneous meiotic resumption of denuded oocytes, SB203580 did not affect GVBD, but it significantly decreased the number of oocytes reaching MII and conversely increased the number of oocytes arrested at MI. These results suggest that p38 MAPK in porcine oocytes becomes active around GVBD, remains active through MI to MII, and has a role in MI-MII transition, and that cumulus p38 MAPK might be involved in FSH-induced meiotic resumption of oocytes. PMID- 15115731 TI - Androgen-regulated genes in the murine epididymis. AB - The epididymis is an androgen-responsive tissue where spermatozoa mature and gain motility. The three major regions of the epididymis, caput, corpus, and cauda, are known to have different functions and exhibit varied gene expression. Specific genes within the different regions of the epididymis have been identified to be under the influence of androgens. The goal of this study was to begin to elucidate the profile of androgen-responsive genes that may be important for sperm maturation using the Affymetrix MGU74Av2 GeneChip oligonucleotide microarray platform. Adult mice (B6/129 strain) were castrated and treated 6 days after castration with two injections of 5 mg of dihydrotestosterone (DHT) or oil over a 48-h period. The mice were killed 48 h later and total RNA was purified from the caput, corpus, and cauda regions of the epididymis. Using GeneSpring 5.0 (Silicon Genetics) software, transcripts were identified that were upregulated 2 fold or more by DHT in the caput (33 transcripts), the corpus (8 transcripts), and the cauda (9 transcripts). PMID- 15115732 TI - Characterization of epididymal epithelial cell-specific gene promoters by in vivo electroporation. AB - The mammalian epididymis plays a critical role in sperm maturation, a function dependent on testicular androgens. However, the function of the initial segment, the most proximal part of the epididymis, is also dependent on luminal factors of testicular origin. Efferent duct ligation (EDL), which prevents luminal testicular fluid from reaching the epididymis, results in changes in gene expression within this region. Cystatin-related epididymal spermatogenic (cres) gene and gamma-glutamyl transpeptidase (GGT) mRNA IV are highly expressed in the initial segment and are regulated by luminal testicular factors. EDL results in decreased expression of both genes. To evaluate these promoters in the context of their native physiological state, an in vivo electroporation procedure was used. Significant differences were observed in vivo compared to previous in vitro results. Whereas two C/EBP sites were necessary for transcriptional activity from a 135-base-pair (bp) cres promoter in vitro, only the 5' site displayed functional activity in the in vivo system. A 135-bp GGT promoter IV construct was sufficient for reporter gene expression in vitro. However, in vivo, substantial expression was not observed until the construct was extended to 530 bp. Three polyoma enhancer activator 3 (PEA3) sites were found to be necessary for in vivo reporter gene expression from this construct. A cis-acting negative regulatory element between -530 and -681 bp was also identified that was not previously recognized in the in vitro studies. These studies demonstrate the utility of in vivo electroporation for elucidating promoter elements that may not be identified when traditional in vitro methods are used. PMID- 15115733 TI - Oligodendrocytes, their progenitors and other neuroglial cells in the aging primate cerebral cortex. AB - In a previous study it was found that with age there is an increase in the frequency of paranodal profiles of myelinated nerve fibers in the cerebral cortex of monkeys. This indicates that there is an increase in the number of internodal myelin segments, and raises the question of whether additional oligodendrocytes are necessary to generate the increased numbers of internodal myelin segments. The present study shows that in layer 4C beta of monkey primary visual cortex there is an age-related increase in the number of oligodendrocytes. When young (4 10 years of age) and old (25-35 years of age) monkeys are compared, the increase is found to be approximately 50%, and it begins in middle age (12-19 years old). It is also shown that although there is no increase in the population of astrocytes in layer 4C beta with age, there appears to be a slight increase in the frequency of microglial cells. As their numbers increase, oligodendrocytes in pairs, rows and groups become more common, which suggests that additional oligodendrocytes are being generated by cell division. Since there is little evidence that mature oligodendrocytes can divide, it is probable that the new oligodendrocytes are generated from progenitor cells which, as many studies have shown, can be labeled by antibodies to NG2, a chondroitin sulfate proteoglycan. By comparing the appearance of these NG2-labeled cells with cells encountered in thin sections of normally prepared tissue, it is shown that the NG2-positive cells have the features of neuroglial cells that were previously described as beta astrocytes. PMID- 15115734 TI - Individual differences in error processing: a review and reanalysis of three event-related fMRI studies using the GO/NOGO task. AB - Three previous studies using the GO/NOGO task were examined to characterize the pattern of functional activation seen during error-related processing. The large sample size (n = 44) also allowed investigation of the influence of individual differences in age, sex, self-reported absentmindedness and reaction speed on the level of activation. Errors were seen to activate a network of regions including the anterior cingulate cortex (ACC), pre-supplementary motor area (pre-SMA), bilateral insula, thalamus and right inferior parietal lobule. Split-half comparisons performed for each of the individual difference variables indicated greater ACC and pre-SMA activation for older subjects while slower responders showed greater activation in the parietal, lateral PFC, insula and ACC regions. Whereas males and females demonstrated equivalent levels of activation in both the ACC and insula, self-reported absentmindedness related to reduced activation in these regions. Our review of the current imaging literature on error-related activation indicates that, despite the use of a variety of other cognitive paradigms, the network of regions identified here is consistent with these previous studies, suggesting that these regions are critical to a 'general' error related response. Furthermore, this response is, in part, influenced by individual differences in both demographic characteristics and behavioural performance. PMID- 15115735 TI - A voxel-based morphometric study to determine individual differences in gray matter density associated with age and cognitive change over time. AB - Voxel-based morphometry (VBM) was used to examine the relation between age and gray matter density cross-sectionally and to study the association between gray matter density and longitudinal decline in performance on cognitive tests in healthy, non-demented elderly individuals. Participants were neuropsychologically tested at baseline and again after 3 years. Thirty-seven subjects (mean age 72.5 years) who showed a decline in cognitive test performance at follow-up were compared with 38 individually matched control subjects (mean age 71.8 years) whose performance did not change over time. Magnetic resonance imaging scans were acquired at follow-up and individual differences in regional gray matter density were examined with VBM. The largest age effects were found in various regions in the prefrontal cortex, the (medial) temporal lobes and the striate cortex. Longitudinal cognitive decline was associated with decreased gray matter density in prefrontal areas, the (medial) temporal lobes and the posterior parietal cortex. These findings suggest that prefrontal and temporal cortical regions are of particular relevance both in aging and age-related cognitive decline in healthy elderly individuals. PMID- 15115736 TI - Perceptual learning on an auditory frequency discrimination task by cats: association with changes in primary auditory cortex. AB - The aim of this study was to determine whether auditory perceptual learning is associated with changes in the frequency organization and/or neuronal response properties of primary auditory cortex (AI). Five out of six cats trained on an 8 kHz frequency discrimination task showed improvements in performance that reflected changes in discriminative capacity. Quantitative measures of the response characteristics and frequency organization of AI revealed that the frequency organization of AI in trained cats did not differ from that in controls, but there was a tendency for neurons with a CF immediately above 8 kHz to have slightly broader tuning in the trained cats than in controls, and neurons in one of these bands had significantly shorter latency. These results are in accord with recent reports that cortical topography in primary visual cortex is unchanged in animals trained on visual discrimination tasks, but are at variance with an earlier report of enlarged representations of training frequencies in AI of monkeys trained on a frequency discrimination task. It is concluded that substantial changes in perceptual discriminative capacity can occur without change in primary cortical topography and with only small changes in neuronal response characteristics. PMID- 15115737 TI - White matter asymmetry in the human brain: a diffusion tensor MRI study. AB - Language ability and handedness are likely to be associated with asymmetry of the cerebral cortex (grey matter) and connectivity (white matter). Grey matter asymmetry, most likely linked to language has been identified with voxel-based morphometry (VBM) using T(1)-weighted images. Differences in white matter obtained with this technique are less consistent, probably due to the relative insensitivity of the T(1) contrast to the ultrastructure of white matter. Furthermore, previous VBM studies failed to find differences related to handedness in either grey or white matter. We revisited these issues and investigated two independent groups of subjects with diffusion-tensor imaging (DTI) for asymmetries in white matter composition. Using voxel-based statistical analyses an asymmetry of the arcuate fascicle was observed, with higher fractional anisotropy in the left hemisphere. In addition, we show differences related to handedness in the white matter underneath the precentral gyrus contralateral to the dominant hand. Remarkably, these findings were very robust, even when investigating small groups of subjects. This highlights the sensitivity of DTI for white matter tissue differences, making it an ideal tool to study small patient populations. PMID- 15115738 TI - Prefrontal selection and medial temporal lobe reactivation in retrieval of short term verbal information. AB - Both the prefrontal cortex (PFC) and medial temporal lobe (MTL) are involved in memory retrieval. Here we distinguish between their roles by manipulating demand for interference resolution and reactivation of maintained information. Subjects were tested on a letter working memory task with distractors. When the memory delay was interrupted with letter distractors, there was higher activity in the middle frontal gyrus at retrieval compared to when the delay was interrupted by number distractors, suggesting the role of the PFC in interference resolution. By contrast, activity in the parahippocampal gyrus did not differ between the two conditions. The two structures were then compared using the dataset of our previous study on memory reactivation, where subjects performed a letter working memory task with arithmetic distractors or non-distractors. The activity in the parahippocampal gyrus at retrieval was higher when distractors interrupted rehearsal than when the intervening task was non-distracting and subjects continued to rehearse, suggesting the role of the MTL in reactivating the stored information. By contrast, the activity in the middle frontal gyrus did not differ between the two conditions. Taking these results together, we have shown the double dissociation between the PFC and MTL in memory retrieval. PMID- 15115739 TI - Unilateral deficits in visual perception and learning after unilateral inferotemporal cortex lesions in macaques. AB - This study adapted the method of partial lesions, combined with controlled fixation, to study the perceptual role of macaque inferotemporal (IT) cortex. Unilateral lesions were made in IT cortex of three monkeys, without section of the corpus callosum, and visual function was tested ipsilateral and contralateral to the lesion. The observed changes were compared to the effects of bilateral lesions of IT cortex in one monkey, the approach used in most previous studies. Unilateral lesions produced far less profound, although more selective, loss on the tested visual abilities than did bilateral lesions. All three monkeys with unilateral lesions showed decreased chromatic sensitivity, but sparing of achromatic sensitivity, and severely disrupted learning and performance of visual matching to sample, and in all cases, the visual loss was contralateral to the site of the lesion. Unexpectedly, the magnitude of the contralateral loss was not increased by later section of the corpus callosum and anterior commissure in one of the monkeys, a lesion that removes interhemispheric input to contralateral from ipsilateral temporal cortex neurons. These results support physiological findings that show that the response of IT cortex neurons is dominated by the contralateral visual field, despite the bilateral activation many IT neurons receive. Comparison to earlier studies of lesions of area V4, which provides input to IT cortex, shows that V4 and IT lesions produce qualitatively different effects. PMID- 15115740 TI - Single cell integration of animate form, motion and location in the superior temporal cortex of the macaque monkey. AB - This study investigated the cellular mechanisms in the anterior part of the superior temporal sulcus (STSa) that underlie the integration of different features of the same visually perceived animate object. Three visual features were systematically manipulated: form, motion and location. In 58% of a population of cells selectively responsive to the sight of a walking agent, the location of the agent significantly influenced the cell's response. The influence of position was often evident in intricate two- and three-way interactions with the factors form and/or motion. For only one of the 31 cells tested, the response could be explained by just a single factor. For all other cells at least two factors, and for half of the cells (52%) all three factors, played a significant role in controlling responses. Our findings support a reformulation of the Ungerleider and Mishkin model, which envisages a subdivision of the visual processing into a ventral 'what' and a dorsal 'where' stream. We demonstrated that at least part of the temporal cortex ('what' stream) makes ample use of visual spatial information. Our findings open up the prospect of a much more elaborate integration of visual properties of animate objects at the single cell level. Such integration may support the comprehension of animals and their actions. PMID- 15115741 TI - Cell type- and region-specific expression of neurogranin mRNA in the cerebral cortex of the macaque monkey. AB - Neurogranin is a postsynaptic substrate for protein kinase C (PKC). It has been identified in the central nervous system, and the expression has been related to postsynaptic plasticity. Using non-radioactive in situ hybridization histochemistry, we investigated whether mRNA expression of neurogranin varied among the cerebral region and cell types. In most areas of the neocortex excluding area OC (the primary visual area), intense signals were observed in the pyramidal cells in layers III, V and VI. In area OC, intense signals were observed in layers IV as well as layers III and VI. We previously showed that intense signals for GAP-43, a presynaptic PKC substrate, were observed in relay neurons of the lateral geniculate nucleus. From this result and the present result in area OC, we conclude that both pre- and postsynaptic PKC substrates (GAP-43 and neurogranin) are abundant in the geniculocortical synapses. In the hippocampus, intense signals were observed in the pyramidal cells in the subiculum. Taken together with our previous study showing intense signals for GAP 43 in Ammon's horn, the result indicates that both PKC substrates are abundant in the connections between neurons in Ammon's horn and in the subiculum. PMID- 15115742 TI - Preferential origin and layer destination of GAD65-GFP cortical interneurons. AB - To identify the origin and track the migratory pathway of specific subpopulations of GABAergic interneurons, we studied tangential migration in a recently developed GAD65-GFP transgenic mouse strain. First, we used immunohistochemical methods to characterize the expression of specific neurochemical markers in the GAD65-GFP neurons. Then, organotypic cultures were used in combination with birth dating studies to determine the time of generation, place of origin and migratory route of these cells. From E14 to E15, the highest density of GAD65-GFP cells was seen in the lower intermediate zone; however, at later stages more GAD65-GFP cells were observed in the subventricular zone. Migratory GAD65-GFP cells express GAD65, but not calretinin or reelin. Surprisingly, only 4% were calbindin immunopositive. At P21, GAD65-GFP cells were found predominantly in layers II-III and expressed calretinin and neuropeptide Y. Remarkably, almost all cholecystokinin-positive but very few parvalbumin-positive neurons expressed GFP. In vitro studies demonstrated that the caudal ganglionic eminence gives rise to a large proportion of GAD65-GFP interneurons and in vivo birth-dating experiments showed that GAD65-GFP interneurons in supragranular layers are born at late embryonic development. Taken together these results support the idea that the destination layer of GABAergic interneurons is closely linked to their place of origin and time of generation. PMID- 15115743 TI - Selective amplification of neocortical neuronal output by fast prepotentials in vivo. AB - Neocortical cells integrate inputs from thousands of presynaptic neurons distributed along their dendritic arbors. Propagation of postsynaptic potentials to the soma is crucial in determining neuronal output. Using intracellular recordings in anesthetized and non-anesthetized, naturally awake and sleeping cats, we found evidence for generation of fast, all-or-none events recorded at the soma in about 20% of regular-spiking and intrinsically-bursting neurons. These events, termed fast prepotentials (FPPs), were suppressed by hyperpolarizing the neurons or by inhibiting synaptic transmission with perfusion of Ca2+-free artificial cerebrospinal fluid. FPPs could be evoked by activation of specific cortical inputs and allowed neurons to fire at more hyperpolarized levels of membrane potentials. Thus, FPPs represent a powerful mechanism to boost the output of neocortical neurons in response to given inputs. We further found evidence for modulation of FPPs generation across the waking-sleep cycle, indicating important changes in the integrative properties of neocortical neurons in different states of vigilance. We suggest that FPPs represent attenuated spikes generated in hot spots of the dendritic arbor and constitute a powerful mechanism to reinforce the functional connections between specific elements of the cortical networks. PMID- 15115744 TI - Expression of serotonin1A and serotonin2A receptors in pyramidal and GABAergic neurons of the rat prefrontal cortex. AB - Serotonergic 5-HT1A and 5-HT2A receptors are abundantly expressed in prefrontal cortex (PFC) and are targets of atypical antipsychotic drugs. They mediate, respectively, inhibitory and excitatory actions of 5-HT. The transcripts for both receptors are largely (approximately 80%) colocalized in rat and mouse PFC, yet their quantitative distribution in pyramidal and GABAergic interneurons is unknown. We used double in situ hybridization histochemistry to estimate the proportion of pyramidal and GABAergic neurons expressing these receptor transcripts in rat PFC. The number of GABAergic interneurons (expressing GAD mRNA) was a 22% of glutamatergic neurons (expressing vGluT1 mRNA, considered as putative pyramidal neurons). 5-HT2A receptor mRNA was present in a large percentage of pyramidal neurons (from 55% in prelimbic cortex to 88% in tenia tecta), except in layer VI, where it was localized only in 30% of those neurons. 5-HT2A receptor mRNA was present in approximately 25% of GAD-containing cells except in layer VI (10%). Likewise, approximately 60% of glutamatergic cells contained the 5-HT1A receptor transcript. We also found that approximately 25% of GAD-expressing cells contained the 5-HT1A receptor mRNA. These data help to clarify the role of 5-HT in prefrontal circuits and shed new light to the cellular elements involved in the action of atypical antipsychotics. PMID- 15115745 TI - Distributed auditory cortical representations are modified when non-musicians are trained at pitch discrimination with 40 Hz amplitude modulated tones. AB - Several functional brain attributes reflecting neocortical activity have been found to be enhanced in musicians compared to non-musicians. Included are the N1m evoked magnetic field, P2 and right-hemispheric N1c auditory evoked potentials, and the source waveform of the magnetically recorded 40 Hz auditory steady state response (SSR). We investigated whether these functional brain attributes measured by EEG are sensitive to neuroplastic remodeling in non-musician subjects. Adult non-musicians were trained for 15 sessions to discriminate small changes in the carrier frequency of 40 Hz amplitude modulated pure tones. P2 and N1c auditory evoked potentials were separated from the SSR by signal processing and found to localize to spatially differentiable sources in the secondary auditory cortex (A2). Training enhanced the P2 bilaterally and the N1c in the right hemisphere where auditory neurons may be specialized for processing of spectral information. The SSR localized to sources in the region of Heschl's gyrus in primary auditory cortex (A1). The amplitude of the SSR (assessed by bivariate T2 in 100 ms moving windows) was not augmented by training although the phase of the response was modified for the trained stimuli. The P2 and N1c enhancements observed here and reported previously in musicians may reflect new tunings on A2 neurons whose establishment and expression are gated by input converging from other regions of the brain. The SSR localizing to A1 was more resistant to remodeling, suggesting that its amplitude enhancement in musicians may be an intrinsic marker for musical skill or an early experience effect. PMID- 15115746 TI - Impaired synaptic plasticity in the surround of perinatally acquired [correction of aquired] dysplasia in rat cerebral cortex. AB - Freeze-lesion induced neocortical dysplasias in rats mimic numerous aspects of human polymicrogyria and are used as a model for the study of developmental migration disorders. Since memory tests have demonstrated learning deficits in rodents with neocortical malformations, we investigated the expression and properties of long-term potentiation (LTP) in neocortical slices from adult freeze-lesioned and control rats. Field potentials, recorded in layer II/III at a distance of 2-3 mm lateral to perinatally induced microgyri, were strongly enhanced following theta-burst stimulation in layer VI (amplitude: 174 +/- 4%) compared to controls (110 +/- 2%). In contrast, in layer IV of the freeze lesioned cortex LTP could not reliably be induced. Histochemical analysis, performed to elucidate the cellular basis of the impaired plasticity, revealed diminished amounts of the GABAA-receptor subunit gamma2 in the paramicrogyral zone, likely representing a diminished GABA-ergic filter, which is thought to prevent LTP induced in layer VI under normal conditions. Cytochrome-oxidase staining after electrophysiological examination disclosed that LTP in layer IV of the freeze-lesioned cortex could only be elicited, when stimulation was applied within a preserved barrel cortex. Our study provides evidence that focal cryolesions during cortical development cause an impaired synaptic plasticity that may underlie learning disabilities. PMID- 15115747 TI - Top-down dendritic input increases the gain of layer 5 pyramidal neurons. AB - The cerebral cortex is organized so that an important component of feedback input from higher to lower cortical areas arrives at the distal apical tufts of pyramidal neurons. Yet, distal inputs are predicted to have much less impact on firing than proximal inputs. Here we show that even weak asynchronous dendritic input to the distal tuft region can significantly increase the gain of layer 5 pyramidal neurons and thereby the output of columns in the primary somatosensory cortex of the rat. Noisy currents injected in ramps at different dendritic locations showed that the initial slope of the frequency-current (f/I) relationship increases with the distance of the current injection from the soma. The increase was due to the interaction of dendritic depolarization with back propagating APs which activated dendritic calcium conductances. Gain increases were accompanied by a change of firing mode from isolated spikes to bursting where the timing of bursts coded the presence of coincident somatic and dendritic inputs. We propose that this dendritic gain modulation and the timing of bursts may serve to associate top-down and bottom-up input on different time scales. PMID- 15115748 TI - Motivation-dependent responses in the human caudate nucleus. AB - Motivation is a complex process that leads to completion or avoidance of a behavior. Past research strongly implicates the basal ganglia in a circuit integral for the control of motivation. Specifically, the human striatum has been shown to process reward information, differentiating between monetary rewards and punishments in recent neuroimaging experiments. It is unclear, however, how the dorsal striatum, particularly the caudate nucleus, responds to changes in the motivational context of a task. Using an event-related design, where participants were given positive and negative feedback upon guessing the value of an unknown card, we manipulated the motivational context of the task by dividing trials into periods of high incentive (where visual feedback indicated monetary rewards and punishments) and low incentive (where visual feedback indicated only accuracy). We found that activity in the caudate nucleus was strongly influenced by the different incentive periods. The hemodynamic response was characterized by a larger rise at the onset of trials and larger differences between positive and negative feedback during periods of high incentive. These results suggest that changes in motivation are capable of modulating basal ganglia activity, and further support an important role for the caudate nucleus in affective processing. PMID- 15115749 TI - Arg343 in human surfactant protein D governs discrimination between glucose and N acetylglucosamine ligands. AB - Surfactant protein D (SP-D), one of the members of the collectin family of C-type lectins, is an important component of pulmonary innate immunity. SP-D binds carbohydrates in a calcium-dependent manner, but the mechanisms governing its ligand recognition specificity are not well understood. SP-D binds glucose (Glc) stronger than N-acetylglucosamine (GlcNAc). Structural superimposition of hSP-D with mannose- binding protein C (MBP-C) complexed with GlcNAc reveals steric clashes between the ligand and the side chain of Arg343 in hSP-D. To test whether Arg343 contributes to Glc > GlcNAc recognition specificity, we constructed a computational model of Arg343-->Val (R343V) mutant hSP-D based on homology with MBP-C. Automated docking of alpha-Me-Glc and alpha-Me-GlcNAc into wild-type hSP-D and the R343V mutant of hSP-D suggests that Arg343 is critical in determining ligand-binding specificity by sterically prohibiting one binding orientation. To empirically test the docking predictions, an R343V mutant recombinant hSP-D was constructed. Inhibition analysis shows that the R343V mutant binds both Glc and GlcNAc with higher affinity than the wild-type protein and that the R343V mutant binds Glc and GlcNAc equally well. These data demonstrate that Arg343 is critical for hSP-D recognition specificity and plays a key role in defining ligand specificity differences between MBP and SP-D. Additionally, our results suggest that the number of binding orientations contributes to monosaccharide binding affinity. PMID- 15115750 TI - Binding patterns of DTR-specific antibodies reveal a glycosylation-conditioned tumor-specific epitope of the epithelial mucin (MUC1). AB - Glycosylation determines essential biological functions of epithelial mucins in health and disease. We report on the influence of glycosylation of the immunodominant DTR motif of MUC1 on its antigenicity. Sets of novel glycopeptides were synthesized that enabled us to examine sole and combined effects of peptide length (number of repeats) and O-glycosylation with GalNAc at the DTR motif on the binding patterns of 22 monoclonal antibodies recognizing this motif. In case of unglycosylated peptides almost all antibodies bound better to multiple MUC1 tandem repeats. Glycosylation at the DTR led to enhanced binding in 11 cases, whereas 10 antibodies were not influenced in binding, and one was inhibited. In nine of the former cases both length and DTR glycosylation were additive in their influence on antibody binding, suggesting that both effects are different. Improved binding to the glycosylated DTR motif was exclusively found with antibodies generated against tumor-derived MUC1. Based on these data a tumor specific MUC1 epitope is defined comprising the ...PDTRP... sequence in a particular conformation essentially determined by O-glycosylation at its threonine with either GalNAcalpha1 or a related short glycan. The results can find application in the field of MUC1-based immunotherapy. PMID- 15115752 TI - G1/S phase cyclin-dependent kinase overexpression perturbs early development and delays tissue-specific differentiation in Xenopus. AB - Cell division and differentiation are largely incompatible but the molecular links between the two processes are poorly understood. Here, we overexpress G1/S phase cyclins and cyclin-dependent kinases in Xenopus embryos to determine their effect on early development and differentiation. Overexpression of cyclin E prior to the midblastula transition (MBT), with or without cdk2, results in a loss of nuclear DNA and subsequent apoptosis at early gastrula stages. By contrast, overexpressed cyclin A2 protein does not affect early development and, when stabilised by binding to cdk2, persists to tailbud stages. Overexpression of cyclin A2/cdk2 in post-MBT embryos results in increased proliferation specifically in the epidermis with concomitant disruption of skin architecture and delay in differentiation. Moreover, ectopic cyclin A2/cdk2 also inhibits differentiation of primary neurons but does not affect muscle. Thus, overexpression of a single G1/S phase cyclin/cdk pair disrupts the balance between division and differentiation in the early vertebrate embryo in a tissue specific manner. PMID- 15115751 TI - The zebrafish iguana locus encodes Dzip1, a novel zinc-finger protein required for proper regulation of Hedgehog signaling. AB - Members of the Hedgehog (Hh) family of intercellular signaling molecules play crucial roles in animal development. Aberrant regulation of Hh signaling in humans causes developmental defects, and leads to various genetic disorders and cancers. We have characterized a novel regulator of Hh signaling through the analysis of the zebrafish midline mutant iguana (igu). Mutations in igu lead to reduced expression of Hh target genes in the ventral neural tube, similar to the phenotype seen in zebrafish mutants known to affect Hh signaling. Contradictory at first sight, igu mutations lead to expanded Hh target gene expression in somites. Genetic and pharmacological analyses revealed that the expression of Hh target genes in igu mutants requires Gli activator function but does not depend on Smoothened function. Our results show that the ability of Gli proteins to activate Hh target gene expression in response to Hh signals is generally reduced in igu mutants both in the neural tube and in somites. Although this reduced Hh signaling activity leads to a loss of Hh target gene expression in the neural tube, the same low levels of Hh signaling appear to be sufficient to activate Hh target genes throughout somites because of different threshold responses to Hh signals. We also show that Hh target gene expression in igu mutants is resistant to increased protein kinase A activity that normally represses Hh signaling. Together, our data indicate that igu mutations impair both the full activation of Gli proteins in response to Hh signals, and the negative regulation of Hh signaling in tissues more distant from the source of Hh. Positional cloning revealed that the igu locus encodes Dzip1, a novel intracellular protein that contains a single zinc-finger protein-protein interaction domain. Overexpression of Igu/Dzip1 proteins suggested that Igu/Dzip1 functions in a permissive way in the Hh signaling pathway. Taken together, our studies show that Igu/Dzip1 functions as a permissive factor that is required for the proper regulation of Hh target genes in response to Hh signals. PMID- 15115753 TI - Hypomorphic expression of Dkk1 in the doubleridge mouse: dose dependence and compensatory interactions with Lrp6. AB - doubleridge is a transgene-induced mouse mutation displaying forelimb postaxial polysyndactyly. We have cloned the doubleridge transgene insertion site and demonstrate that doubleridge acts in cis from a distance of 150 kb to reduce the expression of dickkopf 1 (Dkk1), the secreted Wnt antagonist. Expression of Dkk1 from the doubleridge allele ranges from 35% of wild-type level in E7.0 head to <1% of wild type in E13.5 tail. doubleridge homozygotes and doubleridge/null compound heterozygotes are viable. An allelic series combining the wild-type, doubleridge and null alleles of Dkk1 demonstrates the effect of varying Dkk1 concentration on development of limb, head and vertebrae. Decreasing expression of Dkk1 results in hemivertebral fusions in progressively more anterior positions, with severity increasing from tail kinks to spinal curvature. We demonstrated interaction between Dkk1 and the Wnt coreceptors Lrp5 and Lrp6 by analysis of several types of double mutants. The polydactyly of Dkk1(d/d) mice was corrected by reduced expression of Lrp5 or Lrp6. The posterior digit loss and axial truncation characteristic of Lrp6 null mice was partially corrected by reduction of Dkk1. Similarly, the anterior head truncation characteristic of Dkk1 null mice was rescued by reduction of Lrp6. These compensatory interactions between Dkk1 and Lrp6 demonstrate the importance of correctly balancing positive and negative regulation of Wnt signaling during mammalian development. PMID- 15115754 TI - FGF signaling functions in the hypodermis to regulate fluid balance in C. elegans. AB - Signaling by the Caenorhabditis elegans fibroblast growth factor receptor EGL-15 is activated by LET-756, a fibroblast growth factor, and attenuated by CLR-1, a receptor tyrosine phosphatase. Hyperactive EGL-15 signaling results in a dramatic Clr phenotype characterized by the accumulation of clear fluid within the pseudocoelomic space, suggesting that regulated EGL-15 signaling is essential for fluid homeostasis in C. elegans. To determine the cellular focus of EGL-15 signaling, we identified an enhancer element (e15) within the egl-15 promoter, which is both necessary for the promoter activity and sufficient when duplicated to drive either egl-15 or clr-1 rescue activity. This enhancer drives GFP expression in hypodermal cells. Consistent with this finding, immunofluorescence studies of EGL-15 indicate that EGL-15 is expressed in hypodermal cells, and hypodermal promoters can drive full clr-1 and egl-15 rescue activity. Moreover, a mosaic analysis of mpk-1, which acts downstream of egl-15, suggests that its suppression of Clr (Soc) function is required in the hypodermis. These results suggest that EGL-15 and CLR-1 act in the hypodermis to regulate fluid homeostasis in worms. PMID- 15115755 TI - A Werner syndrome protein homolog affects C. elegans development, growth rate, life span and sensitivity to DNA damage by acting at a DNA damage checkpoint. AB - A Werner syndrome protein homolog in C. elegans (WRN-1) was immunolocalized to the nuclei of germ cells, embryonic cells, and many other cells of larval and adult worms. When wrn-1 expression was inhibited by RNA interference (RNAi), a slight reduction in C. elegans life span was observed, with accompanying signs of premature aging, such as earlier accumulation of lipofuscin and tissue deterioration in the head. In addition, various developmental defects, including small, dumpy, ruptured, transparent body, growth arrest and bag of worms, were induced by RNAi. The frequency of these defects was accentuated by gamma irradiation, implying that they were derived from spontaneous or induced DNA damage. wrn-1(RNAi) worms showed accelerated larval growth irrespective of gamma irradiation, and pre-meiotic germ cells had an abnormal checkpoint response to DNA replication blockage. These observations suggest that WRN-1 acts as a checkpoint protein for DNA damage and replication blockage. This idea is also supported by an accelerated S phase in wrn-1(RNAi) embryonic cells. wrn-1(RNAi) phenotypes similar to those of Werner syndrome, such as premature aging and short stature, suggest wrn-1-deficient C. elegans as a useful model organism for Werner syndrome. PMID- 15115756 TI - The ascidian Mesp gene specifies heart precursor cells. AB - Understanding the molecular basis of heart development is an important research area, because malformation of the cardiovascular system is among the most frequent inborn defects. Although recent research has identified molecules responsible for heart morphogenesis in vertebrates, the initial specification of heart progenitors has not been well characterized. Ascidians provide an appropriate experimental system for exploring this specification mechanism, because the lineage for the juvenile heart is well characterized, with B7.5 cells at the 110-cell stage giving rise to embryonic trunk ventral cells (TVCs) or the juvenile heart progenitors. Here, we show that Cs-Mesp, the sole ortholog of vertebrate Mesp genes in the ascidian Ciona savignyi, is specifically and transiently expressed in the embryonic heart progenitor cells (B7.5 cells). Cs Mesp is essential for the specification of heart precursor cells, in which Nkx, HAND and HAND-like (NoTrlc) genes are expressed. As a result, knockdown of Cs Mesp with specific morpholino antisense oligonucleotides causes failure of the development of the juvenile heart. Together with previous evidence obtained in mice, the present results suggest that a mechanism for heart specification beginning with Mesp through Nkx and HAND is conserved among chordates. PMID- 15115757 TI - Somatic and germline mosaicism in sporadic early-onset Alzheimer's disease. AB - Alzheimer's disease (AD) is the commonest neurodegenerative disease worldwide. Rare familial cases may be caused by mutations in one of three genes-amyloid precursor protein, presenilin-1 and presenilin-2; however, the molecular basis of >99% of AD cases is unknown. Somatic mutation has been considered to be a mechanism that may account for a proportion of sporadic cases of AD, but to date there has been no evidence for this. We now report a sporadic early-onset patient with AD, and show that this individual is a somatic mosaic for a mutation in the presenilin-1 gene, suggesting a novel molecular mechanism for AD. Quantification of the mosaicism demonstrated the degree of mosaicism at 8% in peripheral lymphocytes and 14% in cerebral cortex in the index patient; a clear gene dosage effect on age of presentation and clinical phenotypic presentation is demonstrated. This finding has important implications for the aetiology of sporadic AD, and for other apparently sporadic neurodegenerative diseases such as Parkinson's disease, motor neuron disease and Creutzfeldt-Jakob disease. PMID- 15115758 TI - Direct interaction of FANCD2 with BRCA2 in DNA damage response pathways. AB - Fanconi anaemia (FA) is a chromosomal instability disorder characterized by cellular sensitivity to DNA interstrand crosslinking agents and a high risk of cancer. Six of the eight proteins encoded by the known FA genes form a nuclear complex which is required for the monoubiquitination of the FANCD2 protein. FANCD2 complexes and colocalizes with BRCA1, but its presumptive role in DNA repair has not yet been clearly defined. We used yeast two-hybrid analysis to test for interaction between FANCD2 and 10 proteins involved in homologous recombination repair. FANCD2 did not interact with RAD51, the five RAD51 paralogs, RAD52, RAD54 or DMC1. However, it bound to a highly conserved C terminal site in BRCA2 that also binds FANCG/XRCC9. FANCD2 and BRCA2 can be coimmunoprecipitated from cell extracts of both human and Chinese hamster wild type cells, thus confirming that the interaction occurs in vivo. Formation of nuclear foci of FANCD2 was normal in the BRCA2 mutant CAPAN-1 cells, which indicates that the recruitment of FANCD2 to sites of DNA-repair is independent of wild-type BRCA2 function. FANCD2 colocalized with RAD51 in foci following treatment with mitomycin C or hydroxyurea, and colocalized very tightly with PCNA after treatment with hydroxyurea. These findings suggest that FANCD2 may have a role in the cellular response to stalled replication forks or in the repair of replication-associated double-strand breaks, irrespective of the type of primary DNA lesion. PMID- 15115759 TI - Rescue of lethal molybdenum cofactor deficiency by a biosynthetic precursor from Escherichia coli. AB - Substitution therapies for orphan genetic diseases, including enzyme replacement methods, are frequently hampered by the limited availability of the required therapeutic substance. We describe the isolation of a pterin intermediate from bacteria that was successfully used for the therapy of a hitherto incurable and lethal disease. Molybdenum cofactor (Moco) deficiency is a pleiotropic genetic disorder characterized by the loss of the molybdenum-dependent enzymes sulphite oxidase, xanthine oxidoreductase and aldehyde oxidase due to mutations in Moco biosynthesis genes. An intermediate of this pathway-'precursor Z'-is more stable than the cofactor itself and has an identical structure in all phyla. Thus, it was overproduced in the bacterium Escherichia coli, purified and used to inject precursor Z-deficient knockout mice that display a phenotype which resembles that of the human deficiency state. Precursor Z-substituted mice reach adulthood and fertility. Biochemical analyses further suggest that the described treatment can lead to the alleviation of most symptoms associated with human Moco deficiency. PMID- 15115760 TI - Association of BDNF with anorexia, bulimia and age of onset of weight loss in six European populations. AB - Several genes with an essential role in the regulation of eating behavior and body weight are considered candidates involved in the etiology of eating disorders (ED), but no relevant susceptibility genes with a major effect on anorexia nervosa (AN) or bulimia nervosa (BN) have been identified. Brain-derived neurotrophic factor (BDNF) has been implicated in the regulation of food intake and body weight in rodents. We previously reported a strong association of the Met66 allele of the Val66Met BDNF variant with restricting AN (ANR) and low minimum body mass index in Spanish patients. Another single nucleotide polymorphism located in the promoter region of the BDNF gene (-270C>T) showed lack of association with any ED phenotype. In order to replicate these findings in a larger sample, we performed a case-control study in 1142 Caucasian patients with ED consecutively recruited in six different centers from five European countries (France, Germany, Italy, Spain and UK) participating in the 'Factors in Healthy Eating' project. We have found that the Met66 variant is strongly associated to all ED subtypes (AN, ANR, binge-eating/purging AN and BN), and that the -270C BDNF variant has an effect on BN and late age at onset of weight loss. These are the first two variants associated with the pathophysiology of ED in different populations and support a role for BDNF in the susceptibility to aberrant eating behaviors. PMID- 15115761 TI - The structure of the tau haplotype in controls and in progressive supranuclear palsy. AB - The group of neurodegenerative diseases collectively known as tauopathies are characterized by hallmark lesions consisting of fibrillar aggregates of the microtubule-associated protein, tau (MAPT). Mutations of the tau gene (MAPT) are the cause of frontotemporal dementia with parkinsonism linked to chromosome 17, giving tau a central role in the pathogenic process. The chromosomal region containing MAPT has been shown to evolve into two major haplotypes, H1 and H2, which are defined by linkage disequilibrium (LD) between several polymorphisms over the entire MAPT gene. Studies to date suggest a complete absence of recombination between these two haplotypes. The more common haplotype H1 is over represented in patients with progressive supranuclear palsy (PSP) and corticobasal degeneration. Using single nucleotide polymorphisms, we mapped LD in the regions flanking MAPT and have established the maximum extent of the haplotype block on chromosome 17q21.31 as a region covering approximately 2 Mb. This gene-rich region extends centromerically beyond the corticotrophin releasing hormone receptor 1 gene (CRHR1) to a region of approximately 400 kb, where there is a complete loss of LD. The telomeric end is defined by an approximately 150 kb region just beyond the WNT3 gene. We show that the entire, fully extended H1 haplotype is associated with PSP, which implicates several other genes in addition to MAPT, as candidate pathogenic loci. PMID- 15115762 TI - Ataxin-7 is a subunit of GCN5 histone acetyltransferase-containing complexes. AB - Spinocerebellar ataxia type 7 (SCA7) is a neurodegenerative disorder caused by a CAG repeat expansion in the SCA7 gene leading to elongation of a polyglutamine tract in ataxin-7, a protein of unknown function. A putative ataxin-7 yeast orthologue (SGF73) has been identified recently as a new component of the SAGA (Spt/Ada/Gcn5 acetylase) multisubunit complex, a coactivator required for transcription of a subset of RNA polymerase II-dependent genes. We show here that ataxin-7 is an integral component of the mammalian SAGA-like complexes, the TATA binding protein-free TAF-containing complex (TFTC) and the SPT3/TAF9/GCN5 acetyltransferase complex (STAGA). In agreement, immunoprecipitation of ataxin-7 retained a histone acetyltransferase activity, characteristic for TFTC-like complexes. We further identified a minimal domain in ataxin-7 that is required for interaction with TFTC/STAGA subunits and is conserved highly through evolution, allowing the identification of a SCA7 gene family. We showed that this domain contains a conserved Cys(3)His motif that binds zinc, forming a new zinc binding domain. Finally, polyglutamine expansion in ataxin-7 did not affect its incorporation into TFTC/STAGA complexes purified from SCA7 patient cells. We demonstrate here that ataxin-7 is the human orthologue of the yeast SAGA SGF73 subunit and is a bona fide subunit of the human TFTC-like transcriptional complexes. PMID- 15115763 TI - Gene expression profiles of transcripts in amyloid precursor protein transgenic mice: up-regulation of mitochondrial metabolism and apoptotic genes is an early cellular change in Alzheimer's disease. AB - Alzheimer's disease (AD) is a progressive neurodegenerative disease characterized by the impairment of cognitive functions and by beta amyloid (Abeta) plaques in the cerebral cortex and the hippocampus. Our objective was to determine genes that are critical for cellular changes in AD progression, with particular emphasis on changes early in disease progression. We investigated an established amyloid precursor protein (APP) transgenic mouse model (the Tg2576 mouse model) for gene expression profiles at three stages of disease progression: long before (2 months of age), immediately before (5 months) and after (18 months) the appearance of Abeta plaques. Using cDNA microarray techniques, we measured mRNA levels in 11 283 cDNA clones from the cerebral cortex of Tg2576 mice and age matched wild-type (WT) mice at each of the three time points. This gene expression analysis revealed that the genes related to mitochondrial energy metabolism and apoptosis were up-regulated in 2-month-old Tg2576 mice and that the same genes were up-regulated at 5 and 18 months of age. These microarray results were confirmed using northern blot analysis. Results from in situ hybridization of mitochondrial genes-ATPase-6, heat-shock protein 86 and programmed cell death gene 8-suggest that the granule cells of the hippocampal dentate gyrus and the pyramidal neurons in the hippocampus and the cerebral cortex are up-regulated in Tg2576 mice compared with WT mice. Results from double labeling in situ hybridization suggest that in Tg2576 mice only selective, over expressed neurons with the mitochondrial gene ATPase-6 undergo oxidative damage. These results, therefore, suggest that mitochondrial energy metabolism is impaired by the expression of mutant APP and/or Abeta, and that the up-regulation of mitochondrial genes is a compensatory response. These findings have important implications for understanding the mechanism of Abeta toxicity in AD and for developing therapeutic strategies for AD. PMID- 15115764 TI - Long-range activation of Sox9 in Odd Sex (Ods) mice. AB - The Odd Sex mouse mutation arose in a transgenic line of mice carrying a tyrosinase minigene driven by the dopachrome tautomerase (Dct) promoter region. The minigene integrated 0.98 Mb upstream of Sox9 and was accompanied by a deletion of 134 kb. This mutation causes female to male sex reversal in XX Ods/+ mice, and a characteristic eye phenotype of microphthalmia with cataracts in all mice carrying the transgene. Ods causes sex reversal in the absence of Sry by upregulating Sox9 expression and maintaining a male pattern of Sox9 expression in XX Ods/+ embryonic gonads. This expression, which begins at E11.5, triggers downstream events leading to the formation of a testis. We report here that the 134 kb deletion, in itself, is insufficient to cause sex reversal. We demonstrate that in Ods, the Dct promoter is capable of acting over a distance of 1 Mb to induce inappropriate expression of Sox9 in the retinal pigmented epithelium of the eye, causing the observed microphthalmia. In addition, it induces Sox9 expression in the melanocytes where it causes pigmentation defects. We propose that Ods sex reversal is due to the Dct promoter element interacting with gonad specific enhancer elements to produce the observed male pattern expression of Sox9 in the embryonic gonads. PMID- 15115765 TI - X-Chromosome inactivation ratios affect wild-type MeCP2 expression within mosaic Rett syndrome and Mecp2-/+ mouse brain. AB - Rett syndrome (RTT) is an X-linked neurodevelopmental disorder caused by mutations in MECP2, encoding methyl-CpG-binding protein 2 (MeCP2). The onset of symptoms in RTT is delayed until 6-18 months and 4-6 months in the Mecp2(-/+) mouse model, corresponding to a dynamic and gradual accumulation of MeCP2 expression in individual neurons of the postnatal brain. Because of X chromosome inactivation (XCI), cells within RTT females are mosaic for expression of the heterozygous MECP2 mutation. Using the targeted Mecp2 mouse model, we investigated the effect of Mecp2 mutation on XCI and developmental MeCP2 expression in wild-type (wt)-expressing neurons by quantitative laser scanning cytometry. Mecp2(-/+) female mice exhibited uniform regional distribution of Mecp2 mutant-expressing cells in brain, but unbalanced XCI in the population, favoring expression of the Mecp2 wt allele. Interestingly, MeCP2 expression in Mecp2 wt-expressing cells from Mecp2(-/+) mice was significantly lower than those from Mecp2(+/+) age-matched controls. The negative effect of Mecp2 mutation on wt Mecp2 expression correlated with the percentage of Mecp2 mutant-expressing cells in the cortex. Similar results were observed in two RTT females with identical MECP2 mutations but different XCI ratios. These results demonstrate that Mecp2 mutant neurons affect the development of surrounding neurons in a non-cell autonomous manner and suggest that environmental influences affect the level of MeCP2 expression in wt neurons. These results help in explaining the role of XCI in the pathogenesis of RTT and have important implications in designing therapies for female RTT patients. PMID- 15115766 TI - Progressive decrease in chaperone protein levels in a mouse model of Huntington's disease and induction of stress proteins as a therapeutic approach. AB - The manipulation of chaperone levels has been shown to inhibit aggregation and/or rescue cell death in Saccharomyces cerevisiae, Caenorhabditis elegans, Drosophila melanogaster and cell culture models of Huntington's disease (HD) and other polyglutamine (polyQ) disorders. We show here that a progressive decrease in Hdj1, Hdj2, Hsp70, alphaSGT and betaSGT brain levels likely contributes to disease pathogenesis in the R6/2 mouse model of HD. Despite a predominantly extranuclear location, Hdj1, Hdj2, Hsc70, alphaSGT and betaSGT were found to co localize with nuclear but not with extranuclear aggregates. Quantification of Hdj1 and alphaSGT mRNA levels showed that these do not change and therefore the decrease in protein levels may be a consequence of their sequestration to aggregates, or an increase in protein turnover, possibly as a consequence of their relocation to the nucleus. We have used genetic and pharmacological approaches to assess the therapeutic potential of chaperone manipulation. Ubiquitous overexpression of Hsp70 in the R6/2 mouse (as a result of crossing to Hsp70 transgenics) delays aggregate formation by 1 week, has no effect on the detergent solubility of aggregates and does not alter the course of the neurological phenotype. We used an organotypic slice culture assay to show that pharmacological induction of the heat shock response might be a more useful approach. Radicicol and geldanamycin could both maintain chaperone induction for at least 3 weeks and alter the detergent soluble properties of polyQ aggregates over this time course. PMID- 15115767 TI - Platelet-activating factor-acetylhydrolase and PAF-receptor gene haplotypes in relation to future cardiovascular event in patients with coronary artery disease. AB - Oxidation of low density lipoproteins is an initial step of atherogenesis that generates pro-inflammatory phospholipids, including platelet-activating factor (PAF) and its analogs. PAF is degraded by PAF-acetylhydrolase (PAF-AH), a circulating enzyme having both pro- and anti-inflammatory activities. PAF-AH activity has been postulated to be a risk factor for coronary artery disease (CAD); however, whether PAF-AH has a causal role or is simply a marker of risk is unclear. The aim of this study was to relate the variability of the genes encoding PAF-AH (PLA2G7) and the PAF-receptor (PTAFR) to the risk of CAD and its complications. All polymorphisms located in putatively functional regions were investigated in a prospective cohort of CAD patients (n = 1314) and a group of healthy controls (n = 485). The whole gene variability was investigated in relation to case-control status, prospective cardiovascular outcome and plasma PAF-AH levels by means of haplotype analyses. All analyses indicated an effect of the PLA2G7/A379V polymorphism independent of the other polymorphisms. The V379 allele was less frequent in CAD patients than in controls and was associated with a lower risk of future cardiovascular events, suggesting that this allele might be protective against the development of CAD. The V379 allele was also associated with a weak increase of plasma PAF-AH activity that was unlikely to explain the protective effect of the allele on risk. A more likely interpretation is that the A379V polymorphism might modify the enzyme function towards a more anti atherogenic form. Polymorphisms of the PTAFR gene were not related to any phenotype. PMID- 15115768 TI - GABRD encoding a protein for extra- or peri-synaptic GABAA receptors is a susceptibility locus for generalized epilepsies. AB - A major challenge in understanding complex idiopathic generalized epilepsies has been the characterization of their underlying molecular genetic basis. Here, we report that genetic variation within the GABRD gene, which encodes the GABAA receptor delta subunit, affects GABA current amplitude consistent with a model of polygenic susceptibility to epilepsy in humans. We have found a GABRD Glu177Ala variant which is heterozygously associated with generalized epilepsy with febrile seizures plus. We also report an Arg220His allele in GABRD which is present in the general population. Compared with wild-type receptors, alpha1beta2Sdelta GABAA receptors containing delta Glu177Ala or Arg220His have decreased GABAA receptor current amplitudes. As GABAA receptors mediate neuronal inhibition, the reduced receptor current associated with both variants is likely to be associated with increased neuronal excitability. Since delta subunit-containing receptors localize to extra- or peri-synaptic membranes and are thought to be involved in tonic inhibition, our results suggest that alteration of this process may contribute to the common generalized epilepsies. PMID- 15115769 TI - Purification and characterization of recombinant human prostacyclin synthase. AB - Prostacyclin synthase (PGIS), which catalyzes the conversion of prostaglandin (PG) H(2) to prostacyclin (PGI(2)), is a member of the cytochrome P-450 (P450) superfamily, CYP8A1. To study the enzymatic and protein characteristics of human PGIS, the enzyme was overexpressed in Spodoptera frugiperda 21 (Sf21) cells using the baculovirus expression system. PGIS was expressed in the microsomes of the infected Sf21 cells after culture in 5 microg/ml hematin-supplemented medium for 72 h. The holoenzyme was isolated from the solubilized microsomal fraction by calcium phosphate gel absorption and purified to homogeneity by DEAE-Sepharose and hydroxyapatite column chromatography. The K(m) and V(max) values of the purified human PGIS for PGH(2) were 30 microM and 15 micromol/min/mg of protein at 24 degrees C, respectively. The optical absorption and EPR spectra of the enzyme revealed the characteristics of a low-spin form of P450 in the oxidized state. The carbon monoxide-reduced difference spectrum, however, exhibited a peak at 418 nm rather than 450 nm. The addition of a PGH(2) analogue, U46619, to the enzyme produced an oxygen-ligand type of the difference spectrum with maximum absorption at 407 nm and minimum absorption at 430 nm. Treatment with another PGH(2) analogue, U44069, produced a peak at 387 nm and a trough at 432 nm in the spectrum (Type I), while treatment with tranylcypromine, a PGIS inhibitor, produced a peak at 434 nm and a trough at 412 nm (Type II). A Cys441His mutant of the enzyme possessed no heme-binding ability or enzyme activity. Thus, we succeeded in obtaining a sufficient amount of the purified recombinant human PGIS from infected insect cells for spectral analyses that has high specific activity and the characteristics of a P450, indicating substrate specificity. PMID- 15115770 TI - Correlation between the catalase level in tumor cells and their sensitivity to N beta-alanyl-5-S-glutathionyl-3,4-dihydroxyphenylalanine (5-S-GAD). AB - N-beta-Alanyl-5-S-glutathionyl-3,4-dihydroxyphenylalanine (5-S-GAD) exhibits selective cytotoxicity toward certain human tumor cell lines. 5-S-GAD has been shown to release hydrogen peroxide autonomously. Hydrogen peroxide is converted to water and oxygen by catalase. The purpose of this study is to determine whether or not 5-S-GAD exhibits selective cytotoxicity toward tumor cells with low catalase levels, but not toward ones with high catalase levels. We transfected MDA-MB-435S cells, which are sensitive to 5-S-GAD, with catalase cDNA to establish high catalase producer cells, and then examined their 5-S-GAD sensitivity. Similarly, we repressed catalase expression in T47D cells, which are insensitive to 5-S-GAD, by catalase RNA interference to create low catalase producer cells, and then examined their 5-S-GAD sensitivity. We show that the overexpression of catalase made MDA-MB-435S cells insensitive to 5-S-GAD, whereas the suppression of catalase made T47D cells sensitive to 5-S-GAD. The cellular catalase level was found to be crucial for cell sensitivity to 5-S-GAD. PMID- 15115771 TI - Enzymatic properties of pierisin-1 and its N-terminal domain, a guanine-specific ADP-ribosyltransferase from the cabbage butterfly. AB - The cabbage butterfly, Pieris rapae, produces an ADP-ribosylating cytotoxic protein, pierisin-1. Unlike other ADP-ribosylating toxins, the acceptor site for ADP-ribosylation by pierisin-1 is the N-2 position of guanine bases in DNA. The present study was designed to characterize this novel guanine-specific ADP ribosyltransferase, pierisin-1. The N-terminal polypeptide from Met-1 to Arg-233, but not the C-terminal Ser-234-Met-850 polypeptide, was found to exhibit guanine ADP-ribosyltransferase activity. Trypsin-treated pierisin-1, which is considered to be a "nicked" full-length form composed of associated N- and C-terminal fragments, also demonstrated such activity. Optimum conditions for the N-terminal polypeptide of pierisin-1 were pH 8-10, 37-40 degrees C, in the presence of 100 200 mM NaCl or KCl. Other metal ions such as Ca(2+) or Mg(2+) were not required. Kinetic studies demonstrated potent ADP-ribosyltransferase activity with a K(M) value for NAD of 0.17 mM and k(cat) of 55 per second. Under these optimum conditions, the specific activity of trypsin-treated pierisin-1 was about half (k(cat) = 25 per second). When the conditions were changed to pH 5-7 or 10-20 degrees C, some activity (6-55% or 5-20%, respectively, of that under optimal conditions) of the N-terminal polypeptide was still evident; however, almost all of the trypsin-treated enzyme activity disappeared. This implies the inhibition of the N-terminal enzyme domain by the associated C-terminal fragment. Long-term reactions indicated that a single molecule of pierisin-1 has the capacity to generate more than 10(6) ADP-ribosylated DNA adducts, which could cause the death of a mammalian cell. PMID- 15115772 TI - In vitro phosphorylation of initiation factor 2 alpha (aIF2 alpha) from hyperthermophilic archaeon Pyrococcus horikoshii OT3. AB - Eukaryotic initiation factor 2 (eIF2) is a heterotrimeric protein composed of alpha, beta, and gamma subunits, of which the alpha subunit (eIF2 alpha) plays a crucial role in regulation of protein synthesis through phosphorylation at Ser51. All three subunit genes are conserved in Archaea. To examine the properties of archaeal initiation factor 2 alpha (aIF2 alpha), three genes encoding alpha, beta, and gamma subunits of aIF2 from the hyperthermophilic archaeon Pyrococcus horikoshii OT3 were expressed in Escherichia coli cells, and the resulting proteins, aIF2 alpha, aIF2 beta, and aIF2 gamma, were characterized with reference to the properties of eIF2. aIF2 alpha preferentially interacts with aIF2 gamma, but does not interact with aIF2 beta, which is consistent with data obtained with eIF2, of which eIF2 gamma serves as a core subunit, interacting with eIF2 alpha and eIF2 beta. It was found that aIF2 alpha was, albeit to a lower degree, phosphorylated by double-stranded RNA-dependent protein kinase (hPKR) from human, and a primary target site was suggested to be Ser48 within aIF2 alpha. This finding led us to the search for a putative aIF2 specific kinase gene (PH0512) in the P. horikoshii genome. The gene product Ph0512p unambiguously phosphorylated aIF2 alpha, and Ser48, as in the phosphorylation by hPKR, was suggested to be a target amino acid residue for the PKR homologue Ph0152p in P. horikoshii. These findings suggest that aIF2 alpha, like eIF2 alpha in eukaryotes, plays a role in regulation of the protein synthesis in Archaea through phosphorylation and dephosphorylation. PMID- 15115773 TI - Mass spectrometry of hydrogen/deuterium exchange of Escherichia coli dihydrofolate reductase: effects of loop mutations. AB - To address the effects of single amino acid substitutions on the structural fluctuation of Escherichia coli dihydrofolate reductase (DHFR), hydrogen/deuterium exchange kinetics were investigated at 15 degrees C with wild type and mutant DHFRs at Gly67 (six mutants) and Gly121 (eight mutants) located in two flexible loops, by means of electrospray ionization mass spectrometry. These mutations induced significant changes in the first-order rate constant of proton exchange, k(ex) (0.10-0.27 min(-1)), the number of fast-exchangeable protons, Delta M(o) (164-222 Da), and the number of protons protected from exchange, Delta M(infinity) (15-56 Da), relative to the corresponding values for the wild-type enzyme (k(ex) = 0.18 min(-1), Delta M(o) = 164 Da, and Delta M(infinity) = 50.5 Da). These kinetic parameters were strongly correlated with the volume of introduced amino acids, but partly correlated with adiabatic compressibility (volume fluctuation), stability, and enzymatic activity. These results indicate that the local structure change due to a single amino acid substitution in loop regions is dramatically magnified to affect the structural fluctuation of the whole DHFR molecule, resulting in complicated changes in its stability and function. PMID- 15115774 TI - Characterization of Escherichia coli uridine phosphorylase by single-site mutagenesis. AB - The Escherichia coli udp gene encodes uridine phosphorylase (UP), which catalyzes the reversible phosphorolysis of uridine to uracil and ribose-1-phosphate. The X ray structure of E. coli UP resolved by two different groups produced conflicting results. In order to cast some light on the E. coli UP catalytic site, we mutagenized several residues in UP and measured by RP-HPLC the phosphorolytic activity of the mutant UP proteins in vitro. Mutations Thr94Ala, Phe162Ala, and Tyr195Gly caused a drastic decrease in UP activity. These three residues were suggested to be involved in the nucleoside binding site. However, surprisingly, Tyr195Ala caused a relative increase in enzymatic activity. Both Met197Ala and Met197Ser conserved low activity, suggesting a minor role for this residue in the UP active site. Glu196Ala completely lost UP activity, whereas the more conservative Glu196Asp mutation was still partially active, confirming the importance of maintaining the correct charge in the surroundings of this position. Glu198 was mutated to either Gly, Asp and Gln. All three substitutions caused complete loss of enzymatic activity suggesting an important role of Glu198 both in ribose binding and in interaction with phosphate ions. Arg30Ala and Arg91Ala eliminated UP activity, whereas Arg30Lys and Arg91Lys presented a very low activity, confirming that these residues might interact with and stabilize the phosphate ions. Ile69Ala did not decrease UP activity, whereas His8Ala lowered the activity to about 20%. Both amino acids were suggested to take part in subunit interactions. Our results confirm the structural similarity between E. coli UP and E. coli purine nucleoside phosphorylase (PNP). PMID- 15115775 TI - Immunoelectron microscopy study of polyamines using a newly prepared monoclonal antibody against spermidine: use of a mixture of glutaraldehyde and paraformaldehyde as a cross-linking agent in the preparation of the antigen. AB - We developed a mouse monoclonal antibody (ASPD-19, IgG3 sub-isotype mAb) against spermidine (Spd) conjugated to bovine serum albumin (BSA) using a mixture of glutaraldehyde (GA) and paraformaldehyde (PFA)-sodium borohydride for applications in immunoelectron microscopic studies. The antibody specificity was evaluated by an ELISA binding test simulating the immunocytochemistry (ICC) of tissue sections. The ASPD-19 mAb is highly specific for Spd and Spm, almost the same degree to each, and can distinguish alterations in the chemical structure of other polyamine (PA) analogs, showing less than 3.2% cross-reaction with N(1) acetylspermidine, acetylspermine, or N(8)-acetylspermidine. By an indirect immunoperoxidase method using the ASPD-19 mAb, PA-like immunoreactivities were observed in different tissues fixed with Karnovsky fixative (a mixture of GA and PFA) in combination with borohydride reduction. In contrast, immunoreactivity was very low in tissues when the borohydride reduction step was omitted. The PA-like immunoreaction was completely abolished by the adsorption of the ASPD-19 mAb with 100 microg/ml of Spd or Spm, but was inhibited little or none by other PA-related compounds or amino acids. A light microscopic ICC method using ASPD-19 produced immunostaining of PAs in certain cells in rat tissues with high biosynthetic activities (small intestine, pancreas and spinal cord). A pre-embedding immunoelectron microscopic study using rat spinal cord showed PA immunoreactivity located predominantly on free (polysomes) and attached ribosomes of the rough endoplasmic reticulum (Nissl bodies) in the cytoplasm of motor neurons. These results are in complete agreement with the results obtained by our recent ICC method using another mAb (ASPM-29) produced against GA-conjugated Spm. PMID- 15115776 TI - Characterization and cDNA cloning of monomeric lectins that correspond to the B Chain of a type 2 ribosome-inactivating protein from the bark of Japanese elderberry (Sambucus sieboldiana). AB - Two monomeric lectins, SSA-b-3 and SSA-b-4, were purified from the bark tissue of Japanese elderberry, Sambucus sieboldiana. SDS-PAGE of the purified lectins showed the presence of single bands of 35 and 33 kDa for SSA-b-3 and SSA-b-4, respectively, irrespective of the presence of reducing agent. MS analysis as well as gel filtration of these lectins indicated that they exist mostly as monomeric lectins. Analysis of the N-terminal amino acid sequences of SSA-b-3 and SSA-b-4 yielded an identical sequence, indicating their close structural relationship. Four cDNA clones with extensive homology were obtained from the bark cDNA library and indicated to encode SSA-b-3 or SSA-b-4 from the comparison with the N terminal sequences of these lectins. These clones were classified into two groups, three for SSA-b-3 and one for SSA-b-4, based on the predicted isoelectric points. The amino acid sequences of the encoded polypeptides were almost identical with the B-chain of a type 2 ribosome-inactivating protein from the same bark tissue, sieboldin-b, except for the absence of a small peptide containing a cystein residue, which is critical for the heteromeric dimerization with an A-subunit. Carbohydrate binding specificity and biological activity of these lectins are also reported. PMID- 15115777 TI - 2-Arachidonoylglycerol, an endogenous cannabinoid receptor ligand, induces accelerated production of chemokines in HL-60 cells. AB - 2-Arachidonoylglycerol is an endogenous ligand for the cannabinoid receptors (CB1 and CB2). Previously, we provided evidence that 2-arachidonoylglycerol, but not anandamide (N-arachidonoylethanolamine), is the true natural ligand for the cannabinoid receptors. In the present study, we examined in detail the effects of 2-arachidonoylglycerol on the production of chemokines in human promyelocytic leukemia HL-60 cells. We found that 2-arachidonoylglycerol induced a marked acceleration in the production of interleukin 8. The effect of 2 arachidonoylglycerol was blocked by treatment of the cells with SR144528, a cannabinoid CB2 receptor antagonist, indicating that the effect of 2 arachidonoylglycerol is mediated through the CB2 receptor. Augmented production of interleukin 8 was also observed with CP55940, a synthetic cannabinoid, and an ether-linked analog of 2-arachidonoylglycerol. On the other hand, neither anandamide nor the free arachidonic acid induced the enhanced production of interleukin 8. A similar effect of 2-arachidonoylglycerol was observed in the case of the production of macrophage-chemotactic protein-1. The accelerated production of interleukin 8 by 2-arachidonoylglycerol was observed not only in undifferentiated HL-60 cells, but also in HL-60 cells differentiated into macrophage-like cells. Noticeably, 2-arachidonoylglycerol and lipopolysaccharide acted synergistically to induce the dramatically augmented production of interleukin 8. These results strongly suggest that the CB2 receptor and its physiological ligand, i.e., 2-arachidonoylglycerol, play important regulatory roles such as stimulation of the production of chemokines in inflammatory cells and immune-competent cells. Detailed studies on the cannabinoid receptor system are thus essential to gain a better understanding of the precise regulatory mechanisms of inflammatory reactions and immune responses. PMID- 15115778 TI - Mutational effects on O(6)-methylguanine-DNA methyltransferase from hyperthermophile: contribution of ion-pair network to protein thermostability. AB - Ion pairs have been considered to be general stabilizing factors in hyperthermophilic proteins, but the present experimental data cannot fully explain how ion pairs and ion-pair networks contribute to the stability. In this paper, we show experimental evidence that not all of the internal ion pairs contribute to the thermal and thermodynamic stability, using O(6)-methylguanine DNA methyltransferase from Thermococcus kodakaraensis KOD1 (Tk-MGMT) as a model protein. Of three mutants in which an inter-helical ion pair was disrupted, only one mutant (E93A) was shown to be destabilized. Delta G of E93A was lower by approximately 4 kJ mol(-1) than that of the wild type, and E93A unfolded one order of magnitude faster than did the wild type and other variants. Glu 93 has unique properties in forming an ion-pair network that bridges the N- and C terminal domains and connects three helices in the protein interior. PMID- 15115779 TI - Cloning and expression in Escherichia coli of the D-aspartate oxidase gene from the yeast Cryptococcus humicola and characterization of the recombinant enzyme. AB - The D-aspartate oxidase (DDO) from the yeast Cryptococcus humicola UJ1 (ChDDO) is highly specific to D-aspartate. The gene encoding ChDDO was cloned and expressed in Escherichia coli. Sequence analysis of the ChDDO gene showed that an open reading frame of 1,110 bp interrupted by two introns encodes a protein of 370 amino acids. The deduced amino acid sequence showed an FAD-binding motif and a peroxisomal targeting signal 1 in the N-terminal region and at the C-terminus, respectively, and also the presence of certain catalytically important amino acid residues corresponding to those catalytically important in D-amino acid oxidase (DAO). The sequence exhibited only a moderate identity to human (27.4%) and bovine (28.0%) DDOs, and a rather higher identity to yeast and fungal DAOs (30.4 33.2%). Similarly, phylogenetic analysis showed that ChDDO is more closely related to yeast and fungal DAOs than to mammalian DDOs. The gene expression was regulated at the transcriptional level and specifically induced by the presence of D-aspartate as the sole nitrogen source. ChDDO was expressed in an active form in E. coli to an approximately 5-fold greater extent than in yeast. The purified recombinant enzyme was identical to the native enzyme in physicochemical and catalytic properties. PMID- 15115780 TI - Characterization of cell lines stably expressing human normal or mutated EGFP tagged MC4R. AB - The melanocortin receptor type 4 (MC4-R) is involved in food intake and represents a potential target for the treatment of some forms of obesity. The fluorescent protein EGFP was fused to the wild-type or mutated coding sequence of the human MC4-R. After transfection in HEK 293, clones stably expressing hMC4-R EGFP were selected. Wild-type chimeric hMC4-R was well addressed to the cell membrane as demonstrated using confocal microscopy and displayed the same pharmacological characteristics as native hMC4R. NDP-alpha MSH induced a time dependent internalization of MC4-R that was partially prevented by AgRP. The two mutated chimeric receptors studied here (CTCT-deleted and C271A) showed a high alteration of their response to ligand and were retained inside the cells. In conclusion, we have developed a model of clones stably expressing EGFP-tagged hMC4-R. This is the only such model available to date and it provides a useful tool to follow the trafficking of MC4-R inside living cells. PMID- 15115781 TI - Analysis of autodegradation sites of thermolysin and enhancement of its thermostability by modifying Leu155 at an autodegradation site. AB - The relationship between the autodegradation and thermostability of thermolysin (TLN) was studied. Four autodegradation sites in TLN were identified in the presence of Ca(2+). One of the sites was identified as Gly(154)-Leu(155), and Leu(155) was substituted with various amino acids, X = Ala, Ser, Phe, and Gly, by site-directed mutagenesis. The thermostability at 80 degrees C increased with the amino acid substitutions in the order of Ala>Phe>Ser>Gly>Leu (WT TLN). An additional autodegradation fragment that was not observed with WT TLN appeared for all mutant TLNs examined. The autodegradation site shifted from the Gly(154) Leu(155) bond to the X(155)-Ile(156) one with the mutation at Leu(155). Furthermore, the Ile(164)-Asp(165) bond was recognized newly as an autodegradation site in the mutant TLNs for the production of AF3'. PMID- 15115782 TI - A new recombinant single chain trispecific antibody recruits T lymphocytes to kill CEA (carcinoma embryonic antigen) positive tumor cells in vitro efficiently. AB - Anti-tumor associated antigen (TAA).CD3.CD28 trispecific antibody(TsAb) is able to provide two signals for fully and continuously activation of T lymphocytes and recruit them around tumor cells, presenting an attractive concept in tumor immunotherapy. Here, a new single chain trispecific antibody (scTsAb), named CEA scTsAb, was constructed by fusion of anti-CEA (Carcinoma Embryonic Antigen) single chain antibody (scFv), anti-CD3 scFv and anti-CD28 VH, spaced by polypeptide interlinkers taken from the fragment of constant region (FC) of human IgG and human serum albumin (HSA). It was expressed in Escherichia coli at low temperature (30 degrees C) with up to 50% of the antibody being present in soluble form. After one step of DEAE anion chromatography, the soluble product was sufficiently pure for further in vitro activity assays. First, it was proved that CEA-scTsAb could recognize three antigens (CEA, CD28 and Jurkat cell membrane antigen) specifically and could distinguish antigen positive cells from antigen negative cells in vitro. Then fresh PBMC (peripheral blood mononuclear cells), without being pre-treated by co-stimulatory reagents, such as IL-2 or CD28 mAb, were used as effector cells to test their ability to mediate tumor specific cytolysis of CEA-positive tumor cells, SW1116. It was found by photomicrography that T lymphocytes were attracted to SW1116 cells in the presence of CEA-scTsAb, which resulted in effective cytolysis of tumor cells. As shown by MTT assay, the efficacy of tumor specific cytolysis mediated by CEA scTsAb related to both the quantity and activation of PBMC. At an effector cells/target cells ratio (E/T) of 5, it was proved by dual-color FACS with propidium iodide (PI) and FITC-annexin V that both necrosis and apoptosis of tumor cells were causes of tumor specific cytolysis. In summary, a new single chain trispecific (CEA x CD3 x CD28) antibody was constructed and characterized carefully in this paper and was found to possess functions: (i) to activate T lymphocytes independently of additional co-stimulatory signal, (ii) to attract activated T lymphocytes around CEA-positive tumor cells, (iii) to attack CEA positive tumor cells with recruited T lymphocytes. Because it recognizes a widely distributed tumor antigen (CEA), with moderate molecular weight (about 75 kDa) and a simple production procedure, and is able to mediate a high level of tumor specific cytolysis without any additional co-stimulating reagents, CEA-scTsAb is very promising for the task of immunotherapy in future. PMID- 15115783 TI - Dynamics and reproducibility of a moderately complex sensory-motor response in the medicinal leech. AB - Local bending, a motor response caused by mechanical stimulation of the leech skin, has been shown to be remarkably reproducible, in its initial phase, despite the highly variable firing of motoneurons sustaining it. In this work, the reproducibility of local bending was further analyzed by monitoring it over a longer period of time and by using more intact preparations, in which muscle activation in an entire body segment was studied. Our experiments showed that local bending is a moderately complex motor response, composed of a sequence of four different phases, which were consistently identified in all leeches. During each phase, longitudinal and circular muscles in specific areas of the body segment acted synergistically, being co-activated or co-inhibited depending on their position relative to the stimulation site. Onset and duration of the first phase were reproducible across different trials and different animals as a result of the massive co-activation of excitatory motoneurons sustaining it. The other phases were produced by the inhibition of excitatory and activation of inhibitory motoneurons, and also by the intrinsic relaxation dynamics of leech muscles. As a consequence, their duration and relative timing was variable across different preparations, whereas their order of appearance was conserved. These results suggest that, during local bending, the leech neuromuscular system 1) operates a reduction of its available degrees of freedom, by simultaneously recruiting groups of otherwise antagonistic muscles and large populations of motoneurons; and 2) ensures reliability and effectiveness of this escape reflex, by guaranteeing the reproducibility of its crucial initial phase. PMID- 15115784 TI - Biphasic response to nitric oxide of spinal trigeminal neurons with meningeal input in rat--possible implications for the pathophysiology of headaches. AB - Nitric oxide (NO) is suggested to play a causative role in the pathogenesis of primary headaches. Infusion of NO donors can trigger headache attacks, and products of NO metabolism are found to be increased in the cranial circulation in patients suffering from such headaches. To examine if NO is involved in mediating and maintaining spinal trigeminal neuronal activity, an animal model of meningeal nociception was used. In barbiturate-anesthetized rats, a cranial window was made to expose the parietal dura mater. An access to the medullary brain stem allowed extracellular action potentials to be recorded from neurons in the spinal trigeminal nucleus that received afferent input from the exposed dura. Slow intravenous infusion of the NO donor, sodium nitroprusside (SNP, 50 microg/kg), transiently increased spontaneous activity in a subset of neurons and, with a latency of 50 min, caused a progressive increase in impulse activity across the entire sample of neurons. A similar pattern of delayed activation was seen after topical application of the same dose of SNP onto the exposed medulla. Slow injection of the nonspecific inhibitor of NO synthase, N(omega)-nitro-l-arginine methyl ester (20 mg/kg), reduced the spontaneous activity in all neurons within 15 min. The results suggest that NO can induce delayed, slowly developing activation of central trigeminal neurons and that endogenous release of NO may contribute to the ongoing activity of these neurons. The delayed changes in neuronal activity may include gene expression of pro-nociceptive mediators. These mechanisms may be relevant for the pathogenesis of chronic headaches. PMID- 15115785 TI - Discharge rate of substantia nigra pars reticulata neurons is reduced in non parkinsonian monkeys with apomorphine-induced orofacial dyskinesia. AB - Involuntary movements (dyskinesia) are a common symptom of dopamine-replacement therapy in parkinsonian patients, neuroleptic drug treatment of mental patients, and tic disorders. Levodopa-induced dyskinesia has been shown to be associated with substantial reduction of firing rate in the internal part of the globus pallidus. This study characterizes the changes that occur in the activity of the substantia nigra pars reticulata (SNr) of non-parkinsonian (normal) monkeys with apomorphine (APO)-induced orofacial dyskinesia. We conducted extracellular recordings of SNr neurons of two monkeys before and after induction of orofacial dyskinesia by systemic administration of APO. Involuntary orofacial movements appeared a few minutes after the injections and lasted 20-40 min. Almost all recorded neurons changed their firing rate after APO injection (96%), and most declined (70%). The mean amplitude of decreases was also larger than that of increases (40 vs. 21% of the control rate). Changes in firing pattern were not significant on average. Pairs of SNr neurons were uncorrelated before APO injection, similar to the normal pallidum. However, unlike the increased correlations in the pallidum that accompany parkinsonism, orofacilal dyskinesia in non-parkinsonian monkeys was not associated with changes in correlation between SNr neurons. We conclude that normal monkeys treated with APO can model orofacial dyskinesia and tic disorders that are a consequence of dopaminergic over-activity. These symptoms appear to be more related to reduced firing rate of SNr neurons and thus to disinhibition of their targets, than to changes in pattern and synchronization. PMID- 15115786 TI - Cutaneous inputs can activate the ipsilateral primary motor cortex during bimanual sensory-driven movements in humans. AB - Using transcranial magnetic stimulation (TMS), we examined whether sensory input from a finger affects activity of the ipsilateral primary motor cortex (M1) when human subjects hold a virtual object bimanually and whether this ipsilateral activation varies under different contexts. Subjects used both index fingers to hold two plates, which were subjected to unpredictable pulling loads from torque motors. Loads were delivered in a random sequence to either plate or concurrently to both, although the latter occurred most frequently. Finger forces vertical to the plates and surface electromyographs from the first dorsal interosseous muscles were recorded bilaterally during the task. TMS was sometimes applied over the finger area of the left M1 at variable times relative to load onset to examine cortical excitability. Strength of TMS was set around the active motor threshold of the right finger muscle while subjects waited for loading to the handheld plates. When one plate was singly loaded, the M1 contralateral to the loaded finger was activated, causing automatic force increases in the finger. In addition, the ipsilateral M1 was activated during such loading, associated with transient force increases in the contralateral nonloaded finger. Activations in the ipsilateral M1 were also observed during concurrent loading, when activations were stronger than those following single loading of the contralateral plate. Ipsilateral activations weakened when concurrent loading was less frequent. These results suggest interactions between bilateral sensorimotor cortices during bimanual coordinated movements, with strength varying by context. PMID- 15115787 TI - Cycle period of a network oscillator is independent of membrane potential and spiking activity in individual central pattern generator neurons. AB - Rhythmic motor patterns are thought to arise through the cellular properties and synaptic interactions of neurons in central pattern generator (CPG) circuits. Yet, when examining the CPG underlying the rhythmic escape response of the opisthobranch mollusc, Tritonia diomedea, we found that the cycle period of the fictive swim motor pattern recorded from the isolated nervous system was not altered by changing the resting membrane potential or the level of spiking activity of any of the 3 known CPG cell types: ventral swim interneuron-B (VSI B), the dorsal swim interneurons (DSIs), and cerebral neuron 2 (C2). Furthermore, tonic firing in one or more DSIs or C2 evoked rhythmic bursting that did not differ from the cycle period of the motor pattern evoked by nerve stimulation, regardless of the firing frequency. In contrast, the CPG produced a large range of cycle periods as a function of temperature. The temperature sensitivity of the fictive motor pattern produced by the isolated nervous system was similar to the temperature sensitivity of the swimming behavior produced by the intact animal. Thus, although the CPG is capable of producing a wide range of cycle periods under the influence of temperature, the membrane potentials and spiking activity of the identified CPG neurons do not determine the periodicity of the motor pattern. This suggests that the timing of activity in this network oscillator may be determined by a mechanism that is independent of the membrane potentials and spike rate of its constituent neurons. PMID- 15115788 TI - Pain facilitates tactile processing in human somatosensory cortices. AB - Touch and pain are intimately related modalities. Despite a substantial overlap in their cortical representations interactions between both modalities are largely unknown at the cortical level. We therefore used magnetoencephalography and selective nociceptive cutaneous laser stimulation to investigate the effects of brief painful stimuli on cortical processing of touch. Using a conditioning test stimulus paradigm, our results show that painful conditioning stimuli facilitate processing of tactile test stimuli applied 500 ms later. This facilitation applies to cortical responses later than 40 ms originating from primary (S1) and secondary (S2) somatosensory cortices but not to earlier S1 responses. By contrast, tactile conditioning stimuli yield a decrease of early as well as late responses to tactile test stimuli. Control experiments show that pain-induced facilitation of tactile processing is not restricted to the site of the painful conditioning stimulus, whereas auditory conditioning does not yield a comparable facilitation. Apart from a lack of spatial specificity, the facilitating effect of pain closely resembles attentional effects on cortical processing of tactile stimuli. Thus these findings may represent a physiological correlate of an alerting function of pain as a change in the internal state to prepare for processing signals of particular relevance. PMID- 15115789 TI - Properties of quantal transmission at CA1 synapses. AB - We have used Monte Carlo simulations to understand the generation of quantal responses at the single active zones of CA1 synapses. We constructed a model of AMPA channel activation that accounts for the responses to controlled glutamate application and a model of glutamate diffusion in the synaptic cleft. With no further adjustments to these models, we simulated the response to the release of glutamate from a single vesicle. The predicted response closely matches the rise time of observed responses, which recent measurements show is much faster (<100 micros) than previously thought. The simulations show that initial channel opening is driven by a brief (<100 micros) glutamate spike near the site of vesicle fusion, producing a hotspot of channel activation (diameter: approximately 250 nm) smaller than many synapses. Quantal size therefore depends more strongly on the density of channels than their number, a finding that has important implications for measuring synaptic strength. Recent measurements allow estimation of AMPA receptor density at CA1 synapses. Using this value, our simulations correctly predicts a quantal amplitude of approximately 10 pA. We have also analyzed the properties of excitatory postsynaptic currents (EPSCs) generated by the multivesicular release that can occur during evoked responses. We find that summation is nearly linear and that the existence of multiple narrow peaks in amplitude histograms can be accounted for. It has been unclear how to reconcile the existence of these narrow peaks, which indicate that the variation of quantal amplitude is small (CV < 0.2) with the highly variable amplitude of miniature EPSCs (mEPSCs; CV approximately 0.6). According to one theory, mEPSC variability arises from variation in vesicle glutamate content. However, both our modeling results and recent experimental results indicate that this view cannot account for the observed rise time/amplitude correlation of mEPSCs. In contrast, this correlation and the high mEPSC variability can be accounted for if some mEPSCs are generated by two or more vesicles released with small temporal jitter. We conclude that a broad range of results can be accounted for by simple principles: quantal amplitude (approximately 10 pA) is stereotyped, some mEPSCs are multivesicular at moderate and large synapses, and evoked responses are generated by quasi-linear summation of multiple quanta. PMID- 15115790 TI - Shared and private variability in the auditory cortex. AB - The high variability of cortical sensory responses is often assumed to impose a major constraint on efficient computation. In the auditory cortex, however, response variability can be very low. We have used in vivo whole cell patch-clamp methods to study the trial-to-trial variability of the subthreshold fluctuations in membrane potential underlying tone-evoked responses in the auditory cortex of anesthetized rats. Using methods adapted from classical quantal analysis, we partitioned this subthreshold variability into a private component (which includes synaptic, thermal, and other sources local to the recorded cell) and a shared component arising from network interactions. Here we report that this private component is remarkably small, usually about 1-3 mV, as quantified by the variance divided by the mean of the ensemble of tone-evoked response heights. The shared component can be much larger, and shows more heterogeneity across the population, ranging from about 0 to 10 mV. The remarkable fact that, at least 5 synapses from the auditory periphery, this variability remains so small raises the possibility that the intervening neural circuitry is organized so as to prevent private noise from accumulating as neural signals propagate to the cortex. PMID- 15115791 TI - Do children with focal cerebellar lesions show deficits in shifting attention? AB - More recent findings suggest a possible role of the cerebellum in nonmotor functions. Disability of individuals with cerebellar damage in rapidly shifting attention is one frequently used example to support cerebellar involvement in mental skills. The original proposal was based on findings in five children with chronic surgical lesions of the cerebellum and a young adult with a degenerative disorder. The aim of the present study was to repeat Akshoomoff and Courchesne's initial findings in a larger group of children with focal cerebellar lesions. Ten children with cerebellar lesions and 10 age- and sex-matched controls were tested. Neocerebellar areas were affected in all children with cerebellar damage except one based on detailed analysis of MRI scans. Subjects had to perform a focus and a shift attention task. Two visual and two auditory stimuli were presented in a pseudorandom order. An ellipse and a high-pitched tone were presented less frequently than a circle and a low-pitched tone. Rare stimuli were presented at five different time intervals. In the focus tasks, subjects had to react to the same rare stimulus of one of the two modalities. In the shift task, subjects had to switch between the two rare stimuli. Motor deficits based on reaction times were small in cerebellar children compared with controls. The ability of target detection did not significantly differ in the children with cerebellar lesions compared with the control children in both the focus and the shift attention task. In particular, children with cerebellar damage showed no significant impairment in rapid (<2 s) shifts of attention. The present findings indicate that the cerebellum may be less critical in attention related processes than suggested previously. PMID- 15115792 TI - Neural correlates of the automatic and goal-driven biases in orienting spatial attention. AB - How do stimuli in the environment interact with the goals of observers? We addressed this question by showing that the relevance of an abruptly appearing visual object (cue) changes how observers orient attention toward a subsequent object (target) and how this target is represented in the activity of neurons in the superior colliculus. Initially after the appearance of the cue, attention is driven to its locus. This capture of attention is followed by a second bias in orienting attention, where observers preferentially orient to new locations in the visual scene-an effect called inhibition of return. In the superior colliculus, these two automatic biases in orienting attention were associated with changes in neural activity linked to the appearance of the target-relatively stronger activity linked to the capture of attention and weaker activity linked to inhibition of return. This behavioral pattern changes when the cue predicts the upcoming location of the target-the benefit associated with the capture of attention is enhanced and inhibition of return is reduced. These goal-driven changes in behavior were associated with an increase in pretarget- and target related activity. Taken together, the goals of observers modify stimulus-driven changes in neural activity with both signals represented in the salience maps of the superior colliculi. PMID- 15115794 TI - VR1 receptor activation induces glutamate release and postsynaptic firing in the paraventricular nucleus. AB - Neurons in the paraventricular nucleus (PVN) are important in regulating autonomic function through projections to the brain stem and spinal cord. Although the vanilloid receptors (VR(1)) are present in the PVN, their physiological function is scarcely known. In this study, we determined the role of VR(1) receptors in the regulation of synaptic inputs and the excitability of spinally projecting PVN neurons. Whole cell patch-clamp recordings were performed on the PVN neurons labeled by a retrograde fluorescence tracer injected into the thoracic spinal cord of rats. Capsaicin significantly increased the frequency of glutamatergic miniature excitatory postsynaptic currents (mEPSCs) without changing the amplitude and decay time constant of mEPSCs. On the other hand, capsaicin had no effect on GABAergic miniature inhibitory postsynaptic currents (mIPSCs). The effect of capsaicin on mEPSCs was abolished by a specific VR(1) antagonist, iodo-resiniferatoxin (iodo-RTX), or ruthenium red. Importantly, iodo RTX per se significantly reduced the amplitude of evoked EPSCs and the frequency of mEPSCs. Removal of extracellular Ca(2+), but not Cd(2+) treatment, also eliminated the effect of capsaicin on mEPSCs. Furthermore, capsaicin caused a large increase in the firing rate of PVN neurons, and such an effect was abolished in the presence of ionotropic glutamate receptor antagonists. Additionally, the double-immunofluorescence labeling revealed that all of the VR(1) immunoreactivity was colocalized with a presynaptic marker, synaptophysin, in the PVN. Thus this study provides the first evidence that activation of VR(1) receptors excites preautonomic PVN neurons through selective potentiation of glutamatergic synaptic inputs. Presynaptic VR(1) receptors and endogenous capsaicin-like substances in the PVN may represent a previously unidentified mechanism in hypothalamic regulation of the autonomic nervous system. PMID- 15115793 TI - Interlimb differences in control of movement extent. AB - The current study was designed to examine potential interlimb asymmetries in controlling movement extent. Subjects made repetitive single-joint elbow extension movements while the arm was supported on a horizontal, frictionless, air-jet system. Four targets of 10, 20, 35, and 45 degrees excursions were randomly presented over the course of 150 trials. For both arms, peak tangential hand velocity scaled linearly with movement distance. There was no significant difference between either peak velocities or movement accuracies for the two arms. However, the mechanisms responsible for achieving these velocities and extents were quite distinct for each arm. For the dominant arm, peak tangential finger acceleration varied systematically with movement distance. In contrast, nondominant-arm peak tangential acceleration varied little across targets and, as such, was a poor predictor of movement distance. Instead the velocities of the nondominant arm were determined primarily by variation in the duration of the initial acceleration impulse, which corresponds to the time of peak velocity. These different strategies reflect previously identified mechanisms in controlling movement distance: pulse-height control and pulse-width control. The former is characterized by a variation in peak acceleration and has been associated with preplanning mechanisms. The latter occurs after peak acceleration and has been shown to depend on peripheral sensory feedback. Our findings indicate that the dominant-arm system controls movement extent largely through planning mechanisms that specify pulse-height control, whereas the nondominant system does so largely through feedback mediated pulse-width control. PMID- 15115795 TI - fMRI-adaptation reveals dissociable neural representations of identity and expression in face perception. AB - The distributed model of face processing proposes an anatomical dissociation between brain regions that encode invariant aspects of faces, such as identity, and those that encode changeable aspects of faces, such as expression. We tested for a neuroanatomical dissociation for identity and expression in face perception using a functional MRI (fMRI) adaptation paradigm. Repeating identity across face pairs led to reduced fMRI signal in fusiform cortex and posterior superior temporal sulcus (STS), whereas repeating emotional expression across pairs led to reduced signal in a more anterior region of STS. These results provide neuroanatomical evidence for the distributed model of face processing and highlight a dissociation within right STS between a caudal segment coding identity and a more rostral region coding emotional expression. PMID- 15115796 TI - Phased-array processing for spike discrimination. AB - We present a novel approach for the detection, discrimination, and identification of superimposed neuronal action potentials from multineuronal, multichannel extracellular nerve recordings with low signal-to-noise ratios. The approach uses phased-array processing techniques to identify the spikes from different neurons on the basis of their unique propagation velocities. We evaluated this new approach using simulated electrophysiological data, under conditions that are known to limit the effectiveness of existing spike discrimination techniques. This approach enabled discrimination of simulated spikes from multiple simultaneously active neurons with a high degree of reliability and robustness within the expected range of experimental recording conditions, even in situations where there was a high degree of spike waveform superposition on the recording channels. Moreover, the technique enables the reliable detection and discrimination of spikes recorded with signal-to-noise ratios less than 1. PMID- 15115797 TI - A novel replicating circular DNAzyme. AB - 10-23 DNAzyme has the potential to suppress gene expressions through sequence specific mRNA cleavage. However, the dependence on exogenous delivery limits its applications. The objective of this work is to establish a replicating DNAzyme in bacteria using a single-stranded DNA vector. By cloning the 10-23 DNAzyme into the M13mp18 vector, we constructed two circular DNAzymes, C-Dz7 and C-Dz482, targeting the beta-lactamase mRNA. These circular DNAzymes showed in vitro catalytic efficiencies (kcat/K(M)) of 7.82 x 10(6) and 1.36 x 10(7) M(-1) x min( 1), respectively. Their dependence on divalent metal ions is similar to that found with linear 10-23 DNAzyme. Importantly, the circular DNAzymes were not only capable of replicating in bacteria but also exhibited high activities in inhibiting beta-lactamase and bacterial growth. This study thus provides a novel strategy to produce replicating DNAzymes which may find widespread applications. PMID- 15115799 TI - Mismatching base-pair dependence of the kinetics of DNA-DNA hybridization studied by surface plasmon fluorescence spectroscopy. AB - Two single-stranded DNAs consisting of complementary base pairs except for one mismatching base pair (MM1) can form double-stranded DNA by molecular recognition. This type of duplex is not as stable as that formed by MM0. In order to add to a better understanding of the physical mechanism of the hybridization and dissociation processes at sensor (chip) surfaces, we studied the kinetics of the MM1 hybridization by surface plasmon fluorescence spectroscopy. Target DNA strands labelled with a fluorescent molecule Cy5 at the 5' end and hybridizing with the surface-attached probe DNA can be excited by the strong optical field of a surface plasmon resonance mode. The emitted fluorescence can be detected with high sensitivity. The affinity of a duplex was found to depend on the chemical nature, i.e. G-G, G-T etc., and on the position of the mismatching base pair along the 15mer duplex. PMID- 15115798 TI - Transcription factor binding element detection using functional clustering of mutant expression data. AB - As a powerful tool to reveal gene functions, gene mutation has been used extensively in molecular biology studies. With high throughput technologies, such as DNA microarray, genome-wide gene expression changes can be monitored in mutants. Here we present a simple approach to detect the transcription-factor binding motif using microarray expression data from a mutant in which the relevant transcription factor is deleted. A core part of our approach is clustering of differentially expressed genes based on functional annotations, such as Gene Ontology (GO). We tested our method with eight microarray data sets from the Rosetta Compendium and were able to detect canonical binding motifs for at least four transcription factors. With the support of chromatin IP chip data, we also predict a possible variant of the Swi4 binding motif and recover a core motif for Arg80. Our approach should be readily applicable to microarray experiments using other types of molecular biology techniques, such as conditional knockout/overexpression or RNAi-mediated 'knockdown', to perturb the expression of a transcription factor. Functional clustering included in our approach may also provide new insights into the function of the relevant transcription factor. PMID- 15115800 TI - Meiosis-specific yeast Hop1 protein promotes synapsis of double-stranded DNA helices via the formation of guanine quartets. AB - In most eukaryotes, genetic exchange between paired homologs occurs in the context of a tripartite proteinaceous structure called the synaptonemal complex (SC). Genetic analyses have revealed that the genes encoding SC proteins are vital for meiotic chromosome pairing and recombination. However, the number, nature and/or the mechanism used by SC proteins to align chromosomes are yet to be clearly defined. Here, we show that Saccharomyces cerevisiae Hop1, a component of SC, was able to promote pairing of double-stranded DNA helices containing arrays of mismatched G/G sequences. Significantly, pairing was rapid and robust, independent of homology in the arms flanking the central G/G region, and required four contiguous guanine residues. Furthermore, data from truncated DNA double helices showed that 20 bp on either side of the 8 bp mismatched G/G region was essential for efficient synapsis. Methylation interference indicated that pairing between the two DNA double helices involves G quartets. These results suggest that Hop1 is likely to play a direct role in meiotic chromosome pairing and recombination by its ability to promote synapsis between double-stranded DNA helices containing arrays of G residues. To our knowledge, Hop1 is the first protein shown to promote synapsis of DNA double helices from yeast or any other organism. PMID- 15115801 TI - Whole genome comparisons of serotype 4b and 1/2a strains of the food-borne pathogen Listeria monocytogenes reveal new insights into the core genome components of this species. AB - The genomes of three strains of Listeria monocytogenes that have been associated with food-borne illness in the USA were subjected to whole genome comparative analysis. A total of 51, 97 and 69 strain-specific genes were identified in L.monocytogenes strains F2365 (serotype 4b, cheese isolate), F6854 (serotype 1/2a, frankfurter isolate) and H7858 (serotype 4b, meat isolate), respectively. Eighty-three genes were restricted to serotype 1/2a and 51 to serotype 4b strains. These strain- and serotype-specific genes probably contribute to observed differences in pathogenicity, and the ability of the organisms to survive and grow in their respective environmental niches. The serotype 1/2a specific genes include an operon that encodes the rhamnose biosynthetic pathway that is associated with teichoic acid biosynthesis, as well as operons for five glycosyl transferases and an adenine-specific DNA methyltransferase. A total of 8603 and 105 050 high quality single nucleotide polymorphisms (SNPs) were found on the draft genome sequences of strain H7858 and strain F6854, respectively, when compared with strain F2365. Whole genome comparative analyses revealed that the L.monocytogenes genomes are essentially syntenic, with the majority of genomic differences consisting of phage insertions, transposable elements and SNPs. PMID- 15115802 TI - Patterns of bacterial gene movement. AB - Lateral gene transfer has emerged as an important force in bacterial evolution. A substantial number of genes can be inserted into or deleted from genomes through the process of lateral transfer. In this study, we looked for atypical occurrence of genes among related organisms to detect laterally transferred genes. We have analyzed 50 bacterial complete genomes from nine groups. For each group we use a 16s rRNA phylogeny and a comparison of protein similarity to map gene insertions/deletions onto their species phylogeny. The results reveal that there is poor correlation of genes inserted, deleted, and duplicated with evolutionary branch length. In addition, the numbers of genes inserted, deleted, or duplicated within the same branch are not always correlated with each other. Nor is there any similarity within groups. For example, in the Rhizobiales group, the ratio of insertions to deletions in the evolutionary branch leading to Agrobacterium tumefaciens str. C58 (Cereon) is 0.52, but it is 39.52 for Mesorhizobium loti. Most strikingly, the number of insertions of foreign genes is much larger in the external branches of the trees. These insertions also greatly outnumber the occurrence of deletions, and yet the genome sizes of these bacteria remain roughly constant. This indicates that many of the insertions are specific to each organism and are lost before related species can evolve. Simulations of the process of insertion and deletion, tailored to each phylogeny, support this conclusion. PMID- 15115803 TI - Only a small subset of the horizontally transferred chromosomal genes in Escherichia coli are translated into proteins. AB - Horizontally transferred genes are believed to play a critical role in the divergence of bacterial strains from a common ancestor, but whether all of these genes express functional proteins in the cell remains unknown. Here, we used an integrated LC-based protein identification technology to analyze the proteome of Escherichia coli strain K12 (JM109) and identified 1,480 expressed proteins, which are equivalent to approximately 35% of the total open reading frames predicted in the genome. This subset contained proteins with cellular abundance of several dozens to hundreds of thousands of copies, and included nearly all types of proteins in terms of chemical characteristics, subcellular distribution, and function. Interestingly, the subset also contained 138 of 164 gene products that are currently known to be essential for bacterial viability (84% coverage). However, the subset contained only a very small population (10%) of protein products from genes mapped within K-loops, which are "hot spots" for the integration of foreign DNAs within the K12 genome. On the other hand, these genes in K-loops appeared to be transcribed to RNAs almost as efficiently as the native genes in the bacterial cell as monitored by DNA microarray analysis, raising the possibility that most of the recently acquired foreign genes are inadequate for the translational machinery for the native genes and do not generate functional proteins within the cell. PMID- 15115804 TI - Human urocortin 2, a corticotropin-releasing factor (CRF)2 agonist, and ovine CRF, a CRF1 agonist, differentially alter feeding and motor activity. AB - Two corticotropin-releasing factor (CRF) receptor families have been identified (CRF1 and CRF2). Whereas anxiogenic-like roles for the CRF1 receptor have been identified, behavioral functions of the CRF2 receptor remain obscure. Urocortin 2 (Ucn 2), a CRF-related peptide that selectively binds CRF2 receptors, was recently identified and recognized for its central anorectic properties. The present study tested the hypothesis that the anorexigenic mode of action of Ucn 2 differed from that of ovine CRF (oCRF), a preferential CRF1 receptor agonist. The behavioral effects of intracerebroventricular administration of Ucn 2 were compared with those of oCRF in nondeprived male Wistar rats (n=102). Ucn 2 reduced 6-h food and water intake at doses that did not induce visceral illness (0.1, 1, and 10 microg), as indicated by kaolin intake. Ucn 2 retained its potent anorectic activity in rats receiving a highly palatable cafeteria diet, preferentially reducing intake of carbohydrate (CHO)-rich items while sparing intake of mixed-fat/CHO items. In contrast to Ucn 2, oCRF (10 microg) suppressed 6-h intake of cafeteria diet-fed rats without regard to macronutrient composition. Rather, oCRF most potently suppressed intake of preferred food items. Whereas oCRF had short-onset motor-activating effects, Ucn 2 had nondose dependent, delayed-onset motor-suppressing effects. Thus, central infusion of a CRF2 receptor agonist suppressed intake of both bland and palatable diets without inducing behavioral arousal or malaise, and the profile of anorexigenic effects qualitatively differed from those of a CRF1 receptor agonist. The results suggest the existence of distinct forms of CRF1- and CRF2-mediated anorexia. PMID- 15115805 TI - Dopamine transmission in the human striatum during monetary reward tasks. AB - Previous studies have demonstrated the ability of the [11C]raclopride positron emission tomography (PET) technique to measure behaviorally induced changes in endogenous dopamine transmission in humans. However, these studies have lacked well matched sensorimotor control conditions, making it difficult to know what sensory, cognitive, or motor features contributed to changes in dopaminergic activity. Here we report on [11C]raclopride PET studies in which healthy humans performed card selection tasks for monetary rewards. During separate scans, subjects completed a variable ratio (VR) reward schedule with a 25% reward rate in which they did not know the outcome of their responses in advance, a fixed ratio (FR) 25% reward schedule in which outcomes were fully predictable, and a sensorimotor control (SC) condition involving similar sensory and motor demands but no rewards. Relative to the SC condition, the FR schedule produced only modest increases in dopamine transmission and no decreases relative to the SC condition. In contrast, the VR schedule produced significant increases in dopamine transmission in the left medial caudate nucleus while simultaneously producing significant decreases in other areas of the caudate and putamen. These data indicate: (1) the feasibility of measuring alterations in dopamine transmission even after controlling for sensorimotor features and (2) the complex and regionally specific influence of VR schedules on dopamine transmission. The implications of these results are discussed in relation to conflicting models of dopaminergic functioning arising from studies using electrophysiological and microdialysis techniques in animals. PMID- 15115806 TI - Variations in nucleus accumbens dopamine associated with individual differences in maternal behavior in the rat. AB - Lactating rats exhibit stable individual differences in pup licking/grooming. We used in vivo voltammetry to monitor changes in extracellular dopamine (DA) in the nucleus accumbens (n. Acc) shell of lactating rats interacting with pups and found that (1) the DA signal increased significantly with pup licking/grooming; (2) the onset of such increases preceded pup licking/grooming; and (3) the magnitude and duration of the increase in the DA signal were significantly correlated with the duration of the licking/grooming bout. In females characterized on the basis of behavioral observations as high-licking/grooming mothers, the magnitude of the increase in the DA signal associated with licking/grooming was significantly greater than in low-licking/grooming dams. Dopamine transporter binding in the n. Acc was increased in low-compared with high-licking/grooming mothers. Injection of the selective DA uptake inhibitor GBR 12909 [1-(2-(Bis-(4-fluorophenyl)methoxy)ethyl)-4-(3 phenypropyl)piperazine dihydrochloride] (5 mg/kg, s.c.) increased the DA signal in the n. Acc and pup licking/grooming in low-licking/grooming mothers to levels comparable with those observed in high-licking/grooming dams. Receptor autoradiographic studies showed elevated levels of D1 and D3 receptors in the n. Acc shell region in high licking/grooming dams. These results suggest that high- and low-licking/grooming dams differ in mesolimbic dopaminergic activity associated with mother-pup interactions. Such differences may serve as neural substrates for individual differences in the motivational component of maternal behavior. PMID- 15115807 TI - The role of the frontal pursuit area in learning in smooth pursuit eye movements. AB - The frontal pursuit area (FPA) in the cerebral cortex is part of the circuit for smooth pursuit eye movements. The present paper asks whether the FPA is upstream, downstream, or at the site of learning in pursuit eye movements. Learning was induced by having monkeys repeatedly pursue targets that moved at one speed for 150 msec before changing speed. Single-cell recording showed no consistent correlate of pursuit learning in the responses of FPA neurons. Some neurons showed changes in firing in the same direction as the learning, others showed changes in the opposite direction, and many showed no changes at all. In contrast, the eye movements evoked by electrical stimulation of the FPA showed clear correlates of learning. Learning effects were observed when microstimulation was delivered during the initiation of pursuit and during fixation of a stationary target. In addition, learning caused changes in the degree to which stimulation of the FPA enhanced the eye velocity evoked by brief perturbations of a stationary target. The magnitude of the change in the stimulation-evoked eye movement in each tracking condition was proportional to the size of the eye movement evoked under that condition before learning. We conclude that learning occurs downstream from the FPA, possibly within the cerebellum, and that learning may be related to mechanisms that also control the gain of visual-motor responses on a rapid time scale. PMID- 15115808 TI - Neural correlates of cat odor-induced anxiety in rats: region-specific effects of the benzodiazepine midazolam. AB - Cat odor elicits a profound defensive reaction in rats that is reduced by benzodiazepine drugs. The neural correlates of this phenomenon were investigated here using Fos immunohistochemistry. Rats received either midazolam (0.75 mg/kg, s.c.) or vehicle and were exposed to pieces of a collar that had been worn by a domestic cat or an unworn (dummy) collar. Cat odor caused midazolam-sensitive defensive behavioral responses, including avoidance of collar contact, inhibition of grooming, and prolonged rearing. Cat odor exposure induced Fos expression in the posterior accessory olfactory bulb (glomerular, mitral, and granule cell layers), with granule cell layer activation attenuated by midazolam. High basal Fos expression, and some cat odor-associated Fos expression, was evident in the main olfactory bulb (glomerular cell layer), and midazolam exerted a strong inhibitory effect in this region. Midazolam inhibited Fos expression in key limbic regions involved in pheromone transduction (medial amygdala and bed nucleus of the stria terminalis) and defensive behavior (prelimbic cortex, lateral septum, lateral and medial preoptic areas, and dorsal premammillary nucleus). However, midazolam failed to affect cat odor-related Fos expression in a range of key defense-related sites, including the ventromedial hypothalamic nucleus, paraventricular nucleus of the hypothalamus, periaqueductal gray, and cuneiform nucleus. These results indicate that midazolam exerts a region-specific effect on the neural substrates activated by predator odor, with effects in the lateral septum and dorsal premammillary nucleus likely to be of major importance. These findings also suggest the intriguing hypothesis that cat odor is processed by rats as a "pheromone-like" stimulus. PMID- 15115809 TI - Involvement of monkey inferior colliculus in spatial hearing. AB - The midbrain inferior colliculus (IC) is implicated in coding sound location, but evidence from behaving primates is scarce. Here we report single-unit responses to broadband sounds that were systematically varied within the two-dimensional (2D) frontal hemifield, as well as in sound level, while monkeys fixated a central visual target. Results show that IC neurons are broadly tuned to both sound-source azimuth and level in a way that can be approximated by multiplicative, planar modulation of the firing rate of the cell. In addition, a fraction of neurons also responded to elevation. This tuning, however, was more varied: some neurons were sensitive to a specific elevation; others responded to elevation in a monotonic way. Multiple-linear regression parameters varied from cell to cell, but the only topography encountered was a dorsoventral tonotopy. In a second experiment, we presented sounds from straight ahead while monkeys fixated visual targets at different positions. We found that auditory responses in a fraction of IC cells were weakly, but systematically, modulated by 2D eye position. This modulation was absent in the spontaneous firing rates, again suggesting a multiplicative interaction of acoustic and eye-position inputs. Tuning parameters to sound frequency, location, intensity, and eye position were uncorrelated. On the basis of simulations with a simple neural network model, we suggest that the population of IC cells could encode the head-centered 2D sound location and enable a direct transformation of this signal into the eye-centered topographic motor map of the superior colliculus. Both signals are required to generate rapid eye-head orienting movements toward sounds. PMID- 15115811 TI - Contribution of head shadow and pinna cues to chronic monaural sound localization. AB - Monaurally deaf people lack the binaural acoustic difference cues in sound level and timing that are needed to encode sound location in the horizontal plane (azimuth). It has been proposed that these people therefore rely on spectral pinna cues of their normal ear to localize sounds. However, the acoustic head shadow effect (HSE) might also serve as an azimuth cue, despite its ambiguity when absolute sound levels are unknown. Here, we assess the contribution of either cue in the monaural deaf to two-dimensional (2D) sound localization. In a localization test with randomly interleaved sound levels, we show that all monaurally deaf listeners relied heavily on the HSE, whereas binaural control listeners ignore this cue. However, some monaural listeners responded partly to actual sound-source azimuth, regardless of sound level. We show that these listeners extracted azimuth information from their pinna cues. The better monaural listeners were able to localize azimuth on the basis of spectral cues, the better their ability to also localize sound-source elevation. In a subsequent localization experiment with one fixed sound level, monaural listeners rapidly adopted a strategy on the basis of the HSE. We conclude that monaural spectral cues are not sufficient for adequate 2D sound localization under unfamiliar acoustic conditions. Thus, monaural listeners strongly rely on the ambiguous HSE, which may help them to cope with familiar acoustic environments. PMID- 15115810 TI - Imaging of muscarinic acetylcholine receptor signaling in hippocampal neurons: evidence for phosphorylation-dependent and -independent regulation by G-protein coupled receptor kinases. AB - We used the inositol 1,4,5-trisphosphate (IP3) biosensor, the pleckstrin homology (PH) domain of PLCdelta1 (phospholipase C) tagged with enhanced green fluorescent protein (eGFP-PH(PLCdelta)), to examine muscarinic acetylcholine (mACh) receptor regulation of phospholipase C/IP3 signaling in intact single hippocampal neurons in "real time." Initial experiments produced a pharmacological profile consistent with the presence of a predominant M1 mACh receptor population coupled to the IP3 response. To investigate M1 mACh receptor regulation, neurons were stimulated with approximate EC50 concentrations of the mACh receptor agonist methacholine before (R1) and after (R2) a short (60 sec) exposure to a high concentration of agonist. This resulted in a marked attenuation in the R2 relative to R1 response. Inhibition of endogenous GRK6 (G-protein-coupled receptor kinase) activity, by the introduction of catalytically inactive (K215R)GRK6, partially reversed the attenuation of agonist-induced responsiveness, whereas overexpression of wild type GRK6 increased receptor desensitization. Manipulation of endogenous GRK2 activity through introduction of either wild-type or catalytically inactive GRK2 ((K220R)GRK2) almost completely inhibited agonist-stimulated IP3 production, implying a phosphorylation-independent regulation of M1 mACh receptor signaling, most probably mediated by a GRK2 N-terminal RGS-like (regulator of G-protein signaling) domain interaction with GTP-bound Galpha(q/11). Together, our data suggest a role for both phosphorylation-dependent and -independent regulation of M1 mACh receptors in hippocampal neurons. PMID- 15115812 TI - Functional-neuroanatomic correlates of recollection: implications for models of recognition memory. AB - Recognition decisions can be based on familiarity, the sense that an item was encountered previously (item memory), and on recollection, the conscious recovery of contextual information surrounding a previous encounter with the item (e.g., source memory). Recognition with recollection is thought to depend on multiple mechanisms, including prefrontal "control" processes that guide retrieval and recapitulation mechanisms that reactivate posterior neocortical representations that were present at encoding. However, uncertainty remains regarding the precise nature of prefrontal contributions to recollection and the selectivity of recapitulation to veridical recollection. The present event-related functional magnetic resonance imaging study sought to examine whether regions showing "old new" effects support processes sensitive to recollection success or recollection attempt and whether recapitulation of neocortical representations emerge during veridical recollection as well as during false recognition (i.e., false alarms) or whether false recognition resembles familiarity-based responding. Results revealed that multiple left prefrontal cortical regions were engaged during attempts to recollect previous contextual (source) details, regardless of the nature of the to-be-recollected details and of source recollection outcome (successful vs unsuccessful). Recapitulation effects were observed in regions sensitive to the encoding task, suggesting that veridical recollection entails the reactivation of processes or representations present during encoding. Importantly, in contrast to leading models of recognition memory, false alarms also appeared to be based partially on recollection, as revealed through false recapitulation effects. Implications for neural and cognitive models of recognition are considered. PMID- 15115813 TI - Peripheral treatment with enoxaparin, a low molecular weight heparin, reduces plaques and beta-amyloid accumulation in a mouse model of Alzheimer's disease. AB - We investigated the effect of long-term, peripheral treatment with enoxaparin, a low molecular weight heparin, in transgenic mice overexpressing human amyloid precursor protein(751). Enoxaparin (6 IU per mouse intraperitoneally, three times a week for 6 months) significantly lowered the number and the area occupied by cortical beta-amyloid deposits and the total beta-amyloid (1-40) cortical concentration. Immunocytochemical analysis of glial fibrillary acid protein positive cells showed that enoxaparin markedly reduced the number of activated astrocytes surrounding beta-amyloid deposits. In vitro, the drug dose-dependently attenuated the toxic effect of beta-amyloid on neuronal cells. Enoxaparin dose dependently reduced the ability of beta-amyloid to activate complement and contact systems, two powerful effectors of inflammatory response in AD brain. By reducing the beta-amyloid load and cytotoxicity and proinflammatory activity, enoxaparin offers promise as a tool for slowing the progression of Alzheimer's disease. PMID- 15115814 TI - Neuronal pentraxin 1: a novel mediator of hypoxic-ischemic injury in neonatal brain. AB - Neonatal hypoxic-ischemic brain injury is a major cause of neurological disability and mortality. Its therapy will likely require a greater understanding of the discrete neurotoxic molecular mechanism(s) triggered by hypoxia-ischemia (HI). Here, we investigated the role of neuronal pentraxin 1 (NP1), a member of a newly recognized subfamily of "long pentraxins," in the HI injury cascade. Neonatal brains developed marked infarcts in the ipsilateral cerebral hemisphere at 24 hr and showed significant loss of ipsilateral striatal, cortical, and hippocampal volumes at 7 d after HI compared with the contralateral hemisphere and sham controls. Immunofluorescence analyses revealed elevated neuronal expression of NP1 in the ipsilateral cerebral cortex from 6 hr to 7 d and in the hippocampal CA1 and CA3 regions from 24 hr to 7 d after HI. These same brain areas developed infarcts and terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive cells within 24-48 hr of HI. In primary cortical neurons, NP1 protein was induced >2.5-fold (p < 0.001) after their exposure to hypoxia that caused approximately 30-40% neuronal death. Transfecting cortical neurons with antisense oligodeoxyribonucleotides directed against NP1 mRNA (NP1AS) significantly inhibited (p < 0.01) hypoxia-induced NP1 protein induction and neuronal death (p < 0.001), demonstrating a specific requirement of NP1 in hypoxic neuronal injury. NP1 protein colocalized and coimmunoprecipitated with the fast excitatory AMPA glutamate receptor subunit (GluR1) in primary cortical neurons, and hypoxia induced a time-dependent increase in NP1-GluR1 interactions. NPIAS also protected against AMPA-induced neuronal death (p < 0.05), implicating a role for NP1 in the excitotoxic cascade. Our results show that NP1 induction mediates hypoxic-ischemic injury probably by interacting with and modulating GluR1 and potentially other excitatory glutamate receptors. PMID- 15115815 TI - A synthetic neural cell adhesion molecule mimetic peptide promotes synaptogenesis, enhances presynaptic function, and facilitates memory consolidation. AB - The neural cell adhesion molecule (NCAM) plays a critical role in development and plasticity of the nervous system and is involved in the mechanisms of learning and memory. Here, we show that intracerebroventricular administration of the FG loop (FGL), a synthetic 15 amino acid peptide corresponding to the binding site of NCAM for the fibroblast growth factor receptor 1 (FGFR1), immediately after training rats in fear conditioning or water maze learning, induced a long-lasting improvement of memory. In primary cultures of hippocampal neurons, FGL enhanced the presynaptic function through activation of FGFR1 and promoted synapse formation. These results provide the first evidence for a memory-facilitating effect resulting from a treatment that mimics NCAM function. They suggest that increased efficacy of synaptic transmission and formation of new synapses probably mediate the cognition-enhancing properties displayed by the peptide. PMID- 15115816 TI - Roles of Ca2+, hyperpolarization and cyclic nucleotide-activated channel activation, and actin in temporal synaptic tagging. AB - At crayfish neuromuscular junctions, cAMP increases transmitter released by action potentials by activating two effectors, hyperpolarization and cyclic nucleotide-activated channels (HCNCs) and a separate target that has been tentatively identified as exchange protein activated by cAMP (Epac). Intense electrical activity in the motor neuron induces a long-term facilitation (LTF) of transmitter release in which hyperpolarization from an electrogenic Na+-K+ exchanger activates HCNCs. The coupling of HCNCs to transmission involves actin. After LTF induction, cAMP further increases transmission in an HCNC-independent manner, activating the second target. This relaxation of the requirement for HCNC activation to enhance release is called temporal synaptic tagging. Tagging lasts at least 1 d but develops only in the 10 min period after electrical activity. The HCNCs are activated by the post-tetanic hyperpolarization occurring during this time. Both synaptic tagging and LTF induction depend on presynaptic Ca2+ accumulation during activity; both are blocked by EGTA-AM, and LTF is also prevented by stimulation in a low-[Ca2+] medium. Actin depolymerizers prevent induction of LTF and tagging, with little effect on HCNCs, whose sensitivity to cAMP and HCNC blockers is unaffected by tagging. Enhancement of actin polymerization can rescue tagging from HCNC block, suggesting that actin acts at a step after HCNC activation. These and other recent results suggest a model in which HCNC activation, followed by a process involving actin polymerization, acts cooperatively with [Ca2+] to induce tagging, after which only Epac activation is required for cAMP to further enhance transmission. PMID- 15115817 TI - gemini encodes a zebrafish L-type calcium channel that localizes at sensory hair cell ribbon synapses. AB - L-type Ca2+ channels (LTCCs) drive the bulk of voltage-gated Ca2+ entry in vertebrate inner ear hair cells (HCs) and are essential for mammalian auditory processing. LTCC currents have been implicated in neurotransmitter release at the HC afferent active zone, the ribbon synapse. It is likely that LTCCs play a direct role in vesicle fusion; however, the subcellular localization of the channels in HCs has not been fully resolved. Via positional cloning, we show that mutations in a zebrafish LTCC encoding gene, cav1.3a, underlie the auditory vestibular defects of gemini (gem) circler mutants. gem homozygous receptor mutant HCs display normal cell viability, afferent synaptogenesis, and peripheral innervation, yet exhibit strongly reduced extracellular potentials (approximately 50% of wild-type potentials). Apical FM1-43 uptake, however, is unaffected in gem mutant HCs, suggesting that mechanotransduction channels are functional. Using a Gem-specific antibody, we show that the bulk of Gem/Ca(v)1.3a immunoreactivity in HCs is restricted to basally located focal spots. The number and location of focal spots relative to nerve terminals, and their remarkable ring-shaped structure, which is reminiscent of synaptic dense bodies, are consistent with Gem/Ca(v)1.3a channels clustering at HC ribbon synapses. PMID- 15115818 TI - A novel class of neurons at the trigeminal subnucleus interpolaris/caudalis transition region monitors ocular surface fluid status and modulates tear production. AB - Reflex tears are produced by many conditions, one of which is drying of the ocular surface. Although peripheral neural control of the lacrimal gland is well established, the afferent pathways and properties of central premotor neurons necessary for this reflex are not known. Male rats under barbiturate anesthesia were used to determine whether neurons at the ventral trigeminal subnucleus interpolaris- caudalis (Vi/Vc) transition or the trigeminal subnucleus caudalis cervical cord (Vc/C1) junction region in the lower brainstem were necessary for tears evoked by noxious chemical stimulation (CO2 pulses) or drying of the ocular surface. Both the Vi/Vc transition and Vc/C1 junction regions receive a dense direct projection from corneal nociceptors. Synaptic blockade of the Vi/Vc transition, but not the Vc/C1 junction, by the GABA(A) receptor agonist muscimol inhibited CO2-evoked tears. Glutamate excitation of the Vi/Vc transition, but not the Vc/C1 junction, increased tear volume. Single units recorded at the Vi/Vc transition, but not at the Vc/C1 junction, were inhibited by wetting and excited by drying the ocular surface. Nearly all moisture-sensitive Vi/Vc units displayed an initial inhibitory phase to noxious concentrations of CO2 followed by delayed excitation and displayed an inhibitory surround receptive field from periorbital facial skin. Drying of the ocular surface produced many Fos-positive neurons at the Vi/Vc transition, but not at the Vc/C1 junction. This is the first report of a unique class of moisture-sensitive neurons that exist only at the ventral Vi/Vc transition, and not at more caudal portions of Vc, that may underlie fluid homeostasis of the ocular surface. PMID- 15115819 TI - Trans-endocytosis via spinules in adult rat hippocampus. AB - Locations of a distinctive mode of trans-endocytosis involving dendrites, axons, and glia were quantified through serial section electron microscopy. Short vesicular or long vermiform evaginations emerged from dendrites and axons and were engulfed by presynaptic or neighboring axons, astrocytes, and, surprisingly, a growth cone to form double-membrane structures called spinules. In total, 254 spinules were evaluated in 326 microm(3) of stratum radiatum in area CA1 of mature rat hippocampus. Spinules emerged from spine heads (62%), necks (24%), axons (13%), dendritic shafts (1%), or nonsynaptic protrusions (<1%) and invaginated into axons (approximately 90%), astrocytic processes (approximately 8%), or a growth cone (approximately 1%). Coated pits occurred on the engulfing membrane at the tips of most spinules (69%), and double-membrane structures occurred freely in axonal and astrocytic cytoplasm, suggesting trans-endocytosis. Spinule locations differed among mushroom and thin spines. For mushroom spines, most (84%) of the spinules were engulfed by presynaptic axons, 16% by neighboring axons, and none by astrocytic processes. At thin spines, only 17% of the spinules were engulfed by presynaptic axons, whereas 67% were engulfed by neighboring axons and 14% by astrocytic processes. Spinules engulfed by astrocytic processes support the growing evidence that perisynaptic glia interact directly with synapses at least on thin spines. Spinules with neighboring axons may provide a mechanism for synaptic competition in the mature brain. Trans-endocytosis of spinules by presynaptic axons suggest retrograde signaling or coordinated remodeling of presynaptic and postsynaptic membranes to remove transient perforations and assemble the postsynaptic density of large synapses on mushroom spines. PMID- 15115820 TI - Activity-dependent phosphorylation of tyrosine hydroxylase in dopaminergic neurons of the rat retina. AB - We studied in vivo activity-dependent phosphorylation of tyrosine hydroxylase (TH) in dopaminergic (DA) neurons of the rat retina. TH phosphorylation (TH-P) was evaluated by immunocytochemistry, using antibodies specific for each of three regulated phosphorylation sites. TH synthesis rate was measured by dihydroxyphenylalanine (DOPA) accumulation in the presence of NSD-1015, an inhibitor of aromatic amino acid decarboxylase. TH-P was increased markedly by light or after intraocular injection of GABA(A) and glycine inhibitors. All three phosphospecific antibodies responded similarly to test drugs or light. A 30 min exposure to light increased DOPA accumulation by threefold over that seen after 30 min in darkness. Immunostaining to an anti-panNa channel antibody was found in all parts of the DA neuron. TTX blocked TH-P induced by light or GABA/glycine inhibitors but only in varicosities of the DA axon plexus, not in perikarya or dendrites. Veratridine increased TH-P in all parts of the DA neuron. The distribution of the monoamine vesicular transporter 2 was shown by immunocytochemistry to reside in varicosities of the DA plexus but not in dendrites, indicating that the varicosities are sites of dopamine release. Collectively, these data indicate that, in the retina, dopamine synthesis in varicosities is affected by the spiking activity of retinal neurons, possibly including that of the DA neurons themselves. PMID- 15115821 TI - Early striatal dendrite deficits followed by neuron loss with advanced age in the absence of anterograde cortical brain-derived neurotrophic factor. AB - Brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family, modulates neuronal survival, differentiation, and synaptic function. Reduced BDNF expression in the cortex caused by mutation of the huntingtin gene has been suggested to play a role in the striatal degeneration observed in Huntington's disease. BDNF expression rises dramatically in the cortex during the first few weeks of postnatal life in mice. Previously, it has been impossible to study the specific long-term effects of BDNF absence on CNS structures because of the early postnatal lethality of BDNF-/- mice. Mice harboring a floxed BDNF gene were bred with Emx1(IREScre/+) mice to generate Emx-BDNF(KO) mice that lack cortical BDNF but are viable. Adult Emx-BDNF(KO) mice display a hindlimb clasping phenotype similar to that observed in mouse models of Huntington's disease. The striatum of postnatal Emx-BDNF(KO) mice was reduced in volume compared with controls, and the most abundant neuron type of the striatum, medium spiny neurons (MSNs), had shrunken cell somas, thinner dendrites, and fewer dendritic spines at 35 d of age. Although significant striatal neuron losses were not detected at 35 or 120 d postnatal, 35% of striatal neurons were missing in Emx-BDNF(KO) mice aged beyond 1 year. Thus, cortical BDNF, although not required for the generation or near term survival of MSN, is necessary for normal striatal neuron dendrite morphology during the period when BDNF expression rises in the cortex. Furthermore, a long term in vivo requirement for cortical BDNF in supporting the survival of MSNs is revealed. PMID- 15115822 TI - FE65 constitutes the functional link between the low-density lipoprotein receptor related protein and the amyloid precursor protein. AB - Increasing evidence has implicated the low density lipoprotein receptor-related protein (LRP) and the adaptor protein FE65 in Alzheimer's disease pathogenesis. We have shown previously that LRP mediates beta-amyloid precursor protein (APP) processing and affects amyloid beta-protein and APP secretion and APP-c-terminal fragment generation. Furthermore, LRP mediates APP processing through its intracellular domain. Here, we set out to examine whether this interaction is of direct or indirect nature. Specifically, we asked whether adaptor proteins such as FE65 influence the LRP-mediated effect on APP processing by forming a protein complex. In coimmunoprecipitation experiments, we confirmed the postulated APP FE65 and the LRP-FE65 interaction. However, we also showed an LRP-FE65-APP trimeric complex using pull-down techniques. Because FE65 alters APP processing, we investigated whether this effect is LRP dependent. Indeed, FE65 was only able to increase APP secretion in the presence of LRP. In the absence of LRP, APP secretion was unchanged compared with the LRP knock-out phenotype. Using RNA short interference techniques against FE65, we demonstrated that a reduction in FE65 protein mimics the LRP knock-out phenotype on APP processing. These results clearly demonstrate that FE65 acts as a functional linker between APP and LRP. PMID- 15115823 TI - Bone morphogenetic protein-2 and -4 limit the number of enteric neurons but promote development of a TrkC-expressing neurotrophin-3-dependent subset. AB - The hypothesis that BMPs (bone morphogenetic proteins), which act early in gut morphogenesis, also regulate specification and differentiation in the developing enteric nervous system (ENS) was tested. Expression of BMP-2 and BMP-4, BMPR-IA (BMP receptor subunit), BMPR-IB, and BMPR-II, and the BMP antagonists, noggin, gremlin, chordin, and follistatin was found when neurons first appear in the primordial bowel at embryonic day 12 (E12). Agonists, receptors, and antagonists were detected in separated populations of neural crest- and noncrest-derived cells. When applied to immunopurified E12 ENS precursors, BMP-2 and BMP-4 induced nuclear translocation of phosphorylated Smad-1 (Sma and Mad-related protein). The number of neurons developing from these cells was increased by low concentrations and decreased by high concentrations of BMP-2 or BMP-4. BMPs induced the precocious appearance of TrkC-expressing neurons and their dependence on neurotrophin-3 for survival. BMP-4 interacted with glial cell line-derived neurotrophic factor (GDNF) to enhance neuronal development but limited GDNF driven expansion of the precursor pool. BMPs also promoted development of smooth muscle from mesenchymal cells immunopurified at E12. To determine the physiological significance of these observations, the BMP antagonist noggin was overexpressed in the developing ENS of transgenic mice under the control of the neuron-specific enolase promoter. Neuronal numbers in both enteric plexuses and smooth muscle were increased throughout the postnatal small intestine. These increases were already apparent by E18. In contrast, TrkC-expressing neurons decreased in both plexuses of postnatal noggin-overexpressing animals, again an effect detectable at E18. BMP-2 and/or BMP-4 thus limit the size of the ENS but promote the development of specific subsets of enteric neurons, including those that express TrkC. PMID- 15115824 TI - Neuroendocrine control of larval ecdysis behavior in Drosophila: complex regulation by partially redundant neuropeptides. AB - To complete each molting cycle, insects display a stereotyped sequence of behaviors to shed the remains of the old cuticle. These behavioral routines, as well as other related physiological events, are critical for proper development and are under the control of several neuropeptides. Their correct deployment and concatenation depends on the complex actions and interactions among several peptide hormones: ecdysis triggering hormone (ETH), eclosion hormone (EH), and crustacean cardioactive peptide (CCAP). Numerous theories, some in conflict, have been proposed to define the functional hierarchies by which these regulatory factors operate. Here we use wild-type Drosophila and transgenic flies bearing targeted ablations of either EH or CCAP neurons, or ablations of both together, to reevaluate their roles. Consistent with findings in moths, our results suggest that EH and ETH affect the release of each other via a positive feedback, although ETH can also be released in the absence of EH. We show that EH and ETH both contribute to the air filling of the air ducts (trachea) of the next stage but that EH may play a primary role in this process. We present evidence that EH, whose actions have always been placed upstream of CCAP, may also regulate ecdysis independently of CCAP. Finally, we confirm that flies lacking EH neurons do not ecdyse prematurely when injected with ETH peptides. These findings are surprising and not easily explained by currently available hypotheses. We propose that important additional neuropeptides, and additional interactions between known regulators, contribute to the mechanisms underlying insect ecdysis behaviors. PMID- 15115825 TI - Irradiation of food--helping to ensure food safety. PMID- 15115826 TI - Exchanging kidneys--advances in living-donor transplantation. PMID- 15115827 TI - Translating cancer genomics into clinical oncology. PMID- 15115828 TI - Open to control--new hope for patients with neonatal diabetes. PMID- 15115829 TI - Prediction of survival in diffuse large-B-cell lymphoma based on the expression of six genes. AB - BACKGROUND: Several gene-expression signatures can be used to predict the prognosis in diffuse large-B-cell lymphoma, but the lack of practical tests for a genome-scale analysis has restricted the use of this method. METHODS: We studied 36 genes whose expression had been reported to predict survival in diffuse large B-cell lymphoma. We measured the expression of each of these genes in independent samples of lymphoma from 66 patients by quantitative real-time polymerase-chain reaction analyses and related the results to overall survival. RESULTS: In a univariate analysis, genes were ranked on the basis of their ability to predict survival. The genes that were the strongest predictors were LMO2, BCL6, FN1, CCND2, SCYA3, and BCL2. We developed a multivariate model that was based on the expression of these six genes, and we validated the model in two independent microarray data sets. The model was independent of the International Prognostic Index and added to its predictive power. CONCLUSIONS: Measurement of the expression of six genes is sufficient to predict overall survival in diffuse large-B-cell lymphoma. PMID- 15115830 TI - Activating mutations in the gene encoding the ATP-sensitive potassium-channel subunit Kir6.2 and permanent neonatal diabetes. AB - BACKGROUND: Patients with permanent neonatal diabetes usually present within the first three months of life and require insulin treatment. In most, the cause is unknown. Because ATP-sensitive potassium (K(ATP)) channels mediate glucose stimulated insulin secretion from the pancreatic beta cells, we hypothesized that activating mutations in the gene encoding the Kir6.2 subunit of this channel (KCNJ11) cause neonatal diabetes. METHODS: We sequenced the KCNJ11 gene in 29 patients with permanent neonatal diabetes. The insulin secretory response to intravenous glucagon, glucose, and the sulfonylurea tolbutamide was assessed in patients who had mutations in the gene. RESULTS: Six novel, heterozygous missense mutations were identified in 10 of the 29 patients. In two patients the diabetes was familial, and in eight it arose from a spontaneous mutation. Their neonatal diabetes was characterized by ketoacidosis or marked hyperglycemia and was treated with insulin. Patients did not secrete insulin in response to glucose or glucagon but did secrete insulin in response to tolbutamide. Four of the patients also had severe developmental delay and muscle weakness; three of them also had epilepsy and mild dysmorphic features. When the most common mutation in Kir6.2 was coexpressed with sulfonylurea receptor 1 in Xenopus laevis oocytes, the ability of ATP to block mutant K(ATP) channels was greatly reduced. CONCLUSIONS: Heterozygous activating mutations in the gene encoding Kir6.2 cause permanent neonatal diabetes and may also be associated with developmental delay, muscle weakness, and epilepsy. Identification of the genetic cause of permanent neonatal diabetes may facilitate the treatment of this disease with sulfonylureas. PMID- 15115832 TI - Fibromuscular dysplasia. PMID- 15115831 TI - Triple-nucleoside regimens versus efavirenz-containing regimens for the initial treatment of HIV-1 infection. AB - BACKGROUND: Regimens containing three nucleoside reverse-transcriptase inhibitors offer an alternative to regimens containing nonnucleoside reverse-transcriptase inhibitors or protease inhibitors for the initial treatment of human immunodeficiency virus type 1 (HIV-1) infection, but data from direct comparisons are limited. METHODS: This randomized, double-blind study involved three antiretroviral regimens for the initial treatment of subjects infected with HIV 1: zidovudine-lamivudine-abacavir, zidovudine-lamivudine plus efavirenz, and zidovudine-lamivudine-abacavir plus efavirenz. RESULTS: We enrolled a total of 1147 subjects with a mean baseline HIV-1 RNA level of 4.85 log10 (71,434) copies per milliliter and a mean CD4 cell count of 238 per cubic millimeter were enrolled. A scheduled review by the data and safety monitoring board with the use of prespecified stopping boundaries led to a recommendation to stop the triple nucleoside group and to present the results in the triple-nucleoside group in comparison with pooled data from the efavirenz groups. After a median follow-up of 32 weeks, 82 of 382 subjects in the triple-nucleoside group (21 percent) and 85 of 765 of those in the combined efavirenz groups (11 percent) had virologic failure; the time to virologic failure was significantly shorter in the triple nucleoside group (P<0.001). This difference was observed regardless of the pretreatment HIV-1 RNA stratum (at least 100,000 copies per milliliter or below this level; P< or =0.001 for both comparisons). Changes in the CD4 cell count and the incidence of grade 3 or grade 4 adverse events did not differ significantly between the groups. CONCLUSIONS: In this trial of the initial treatment of HIV-1 infection, the triple-nucleoside combination of abacavir, zidovudine, and lamivudine was virologically inferior to a regimen containing efavirenz and two or three nucleosides. PMID- 15115834 TI - Images in clinical medicine. The Mount Fuji sign. PMID- 15115833 TI - Where does HIV live? PMID- 15115835 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 14-2004. A 66-year-old man with progressive neurologic deficits. PMID- 15115836 TI - The role of irradiation in food safety. PMID- 15115837 TI - Clues for new therapeutics in osteoporosis. PMID- 15115838 TI - Soluble TREM-1 and the diagnosis of pneumonia. PMID- 15115839 TI - Circulating IGF-I deficiency and inactivation of the acid-labile subunit gene. PMID- 15115840 TI - The Budd-Chiari syndrome. PMID- 15115841 TI - Abortion, health, and the law. PMID- 15115842 TI - Paying physicians for high-quality care. PMID- 15115843 TI - Refractory thrombocytopenia despite treatment for rattlesnake envenomation. PMID- 15115844 TI - State initiatives to control Medicaid drug costs. PMID- 15115845 TI - Minor transplacental passage of fondaparinux in vivo. PMID- 15115846 TI - Images in clinical medicine. Pneumoperitoneum. PMID- 15115847 TI - From womb the bell tolls. PMID- 15115850 TI - Methods for genetic dissection of complex traits. AB - In this Perspective, the author responds to a recent SAGE KE Perspective. An alternative view of the benefits of quantitative trait loci mapping as a way to understand the molecular mechanisms of life-span determination is presented. PMID- 15115848 TI - Bad to the bone. PMID- 15115851 TI - Cluster analysis of water molecules in alanine racemase and their putative structural role. AB - Conservation of water molecules was identified by a cluster analysis of seven crystal structures of alanine racemase from Bacillus stearothermophilus. A total of 47 clusters of consensus water sites were determined and found to be highly localized, as indicated by their low mobilities. These clusters are located in the region of the active sites as well as at the interface between the N-terminal domain (the alpha/beta-barrel) of the first monomer and the C-terminal domain of the second monomer. The clusters located at the dimer interface form extensive hydrogen-bonding networks linked to the protein backbone. These water-mediated hydrogen bonds, and also all hydrogen-bonding interactions at the dimer interface, were monitored during a 2 ns molecular dynamics simulation and showed that when the inhibitor propionate was bound to the enzyme, some of these interactions were disrupted. The data we present here indicate that the consensus water sites identified at the interface between the two monomers of alanine racemase may play a structural role, which is to maintain and stabilize the alanine racemase dimer. A second role might be to supply the active site continuously with water molecules in order to allow rapid equilibration of active site protons with the solvent. PMID- 15115852 TI - Thermal stabilization of penicillolysin, a thermolabile 19 kDa Zn2+-protease, obtained by site-directed mutagenesis. AB - Penicillolysin is a member of the clan MX and the family of M35 proteases. The enzyme is a thermolabile Zn(2+)- protease from Penicillium citrinum with a unique substrate profile. We expressed recombinant penicillolysin in Aspergillus oryzae and generated several site-directed mutants, R33E/E60R, A167E and T81P, with the intention of exploring thermal stabilization of this protein. We based our choice of mutations on the structures of homologous thermally stable enzymes, deuterolysin (EC 3.4.24.39) from A.oryzae and a peptidyl-Lys metallopeptidase (GfMEP) from the edible mushroom Grifora frondsa. The resulting mutant proteins exhibited comparable catalytic efficiency to the wild-type enzyme and some showed a higher tolerance to temperature. PMID- 15115853 TI - Directed evolution of an anti-carcinoembryonic antigen scFv with a 4-day monovalent dissociation half-time at 37 degrees C. AB - An scFv has been engineered to bind carcinoembryonic antigen (CEA) with a dissociation half-time >4 days at 37 degrees C. Two mutations responsible for this affinity increase were isolated by screening yeast surface-displayed mutant libraries by flow cytometry. Soluble expression of the mutant scFv in a yeast secretion system was increased 100-fold by screening mutant libraries for improved yeast surface display level. This scFv will be useful as a limiting case for evaluating the significance of affinity in tumor targeting to non internalizing antigens. PMID- 15115854 TI - Feature-based prediction of non-classical and leaderless protein secretion. AB - We present a sequence-based method, SecretomeP, for the prediction of mammalian secretory proteins targeted to the non-classical secretory pathway, i.e. proteins without an N-terminal signal peptide. So far only a limited number of proteins have been shown experimentally to enter the non-classical secretory pathway. These are mainly fibroblast growth factors, interleukins and galectins found in the extracellular matrix. We have discovered that certain pathway-independent features are shared among secreted proteins. The method presented here is also capable of predicting (signal peptide-containing) secretory proteins where only the mature part of the protein has been annotated or cases where the signal peptide remains uncleaved. By scanning the entire human proteome we identified new proteins potentially undergoing non-classical secretion. Predictions can be made at http://www.cbs.dtu.dk/services/SecretomeP. PMID- 15115855 TI - Sleep disordered breathing and the outcome of stroke. PMID- 15115856 TI - Alpha 1-antitrypsin: more than just deficiency. PMID- 15115857 TI - 2004 update of BTS pneumonia guidelines: what's new? PMID- 15115859 TI - Effect of upper airway obstruction in acute stroke on functional outcome at 6 months. AB - BACKGROUND: The aim of this study was to determine whether upper airway obstruction occurring within the first 24 hours of stroke onset has an effect on outcome following stroke at 6 months. Traditional definitions used for obstructive sleep apnoea (OSA) are arbitrary and may not apply in the acute stroke setting, so a further aim of the study was to redefine respiratory events and to assess their impact on outcome. METHODS: 120 patients with acute stroke underwent a sleep study within 24 hours of onset to determine the severity of upper airway obstruction (respiratory disturbance index, RDI-total study). Stroke severity (Scandinavian Stroke Scale, SSS) and disability (Barthel score) were also recorded. Each patient was subsequently followed up at 6 months to determine morbidity and mortality. RESULTS: Death was independently associated with SSS (OR (95% CI) 0.92 (0.88 to 0.95), p<0.00001) and RDI-total study (OR (95% CI) 1.07 (1.03 to 1.12), p<0.01). The Barthel index was independently predicted by SSS (p = 0.0001; r = 0.259; 95% CI 0.191 to 0.327) and minimum oxygen saturation during the night (p = 0.037; r = 0.16; 95% CI 0.006 to 0.184). The mean length of the respiratory event most significantly associated with death at 6 months was 15 seconds (sensitivity 0.625, specificity 0.525) using ROC curve analysis. CONCLUSION: The severity of upper airway obstruction appears to be associated with a worse functional outcome following stroke, increasing the likelihood of death and dependency. Longer respiratory events appear to have a greater effect. These data suggest that long term outcome might be improved by reducing upper airway obstruction in acute stroke. PMID- 15115860 TI - Failure of sputum eosinophilia after eotaxin inhalation in asthma. AB - BACKGROUND: Eotaxin is a chemokine specific for eosinophils and may play an important role in eosinophil recruitment in asthma. The effects of eotaxin inhalation on sputum and blood eosinophils, exhaled nitric oxide (NO), and bronchial responsiveness were determined. METHODS: Eotaxin was administered by nebulisation to asthma patients in three studies: (1) an open dose finding study with eotaxin (5, 10 and 20 microg) to two asthmatic subjects; (2) a randomised placebo controlled study with 20 microg eotaxin to five asthmatic subjects and five normal volunteers; and (3) a randomised placebo controlled study with 40 microg eotaxin to nine asthmatics. Forced expiratory volume in 1 second (FEV(1)), exhaled NO, and blood eosinophils were measured before and hourly for 5 hours after nebulisation and at 24 and 72 hours. Methacholine bronchial challenge and sputum induction were performed before and at 5, 24, and 72 hours after nebulisation. RESULTS: In the two placebo controlled studies there was no change in sputum eosinophil count and sputum eosinophilic cationic protein concentration after eotaxin inhalation compared with placebo. FEV(1), exhaled NO, and methacholine PC(20) did not change. However, high dose eotaxin (40 microg) induced an increase in sputum neutrophil count compared with placebo (p<0.05). CONCLUSIONS: Inhaled eotaxin up to 40 microg induced no changes in sputum eosinophil count but at 40 microg it increased the sputum neutrophil count. The significance of this finding is unknown. PMID- 15115861 TI - Relationship between socioeconomic status and asthma: a longitudinal cohort study. AB - BACKGROUND: There is conflicting information about the relationship between asthma and socioeconomic status, with different studies reporting no, positive, or inverse associations. Most of these studies have been cross sectional in design and have relied on subjective markers of asthma such as symptoms of wheeze. Many have been unable to control adequately for potential confounding factors. METHODS: We report a prospective cohort study of approximately 1000 individuals born in Dunedin, New Zealand in 1972-3. This sample has been assessed regularly throughout childhood and into adulthood, with detailed information collected on asthma symptoms, lung function, airway responsiveness, and atopy. The prevalence of these in relation to measures of socioeconomic status were analysed with and without controls for potential confounding influences including parental history of asthma, smoking, breast feeding, and birth order using cross sectional time series models. RESULTS: No consistent association was found between childhood or adult socioeconomic status and asthma prevalence, lung function, or airway responsiveness at any age. Having asthma made no difference to educational attainment or socioeconomic status by age 26. There were trends to increased atopy in children from higher socioeconomic status families consistent with previous reports. CONCLUSIONS: Socioeconomic status in childhood had no significant impact on the prevalence of asthma in this New Zealand born cohort. Generalisation of these results to other societies should be done with caution, but our results suggest that the previously reported associations may be due to confounding. PMID- 15115863 TI - Do farming exposures cause or prevent asthma? Results from a study of adult Norwegian farmers. AB - BACKGROUND: A protective effect of endotoxin exposure on atopy and asthma in farmers' children has been postulated. Studies of adult farmers have shown conflicting results but often lack exposure data. The prevalence of asthma in farmers with different exposure levels to microbial agents and irritant gases was compared. METHODS: Atopy was defined as a positive response to multiple radioallergosorbent tests (RAST) with a panel of 10 common respiratory allergens, and asthma was ascertained by a questionnaire using a stratified sample (n = 2169) of a farming population from south-eastern Norway. Exposure of farmers to total dust, fungal spores, bacteria, endotoxins, and ammonia was assessed by exposure measurements. RESULTS: The prevalence of asthma was 3.7% for physician diagnosed asthma and 2.7% for current asthma. The prevalence of atopy was 14%, but most asthmatic subjects were non-atopic (80%). Compared with farmers without livestock, (1) asthma was significantly higher in cattle farmers (OR(adj) 1.8, 95% CI 1.1 to 2.8) and pig farmers (OR(adj) 1.6, 95% CI 1.0 to 2.5), (2) non atopic asthma was significantly higher in pig farmers (OR(adj) 2.0, 95% CI 1.2 to 3.3) and in farmers with two or more types of livestock (OR(adj) 1.9, 95% CI 1.1 to 3.3), and (3) atopic asthma was less common in farmers with two or more types of livestock (OR(adj) 0.32, 95% CI 0.11 to 0.97). Exposure to endotoxins, fungal spores, and ammonia was positively associated with non-atopic asthma and negatively associated with atopic asthma. No associations were found with atopy. CONCLUSIONS: Exposure to endotoxins and fungal spores appears to have a protective effect on atopic asthma but may induce non-atopic asthma in farmers. PMID- 15115864 TI - Effect of exacerbations on quality of life in patients with chronic obstructive pulmonary disease: a 2 year follow up study. AB - BACKGROUND: A study was undertaken to evaluate exacerbations and their impact on the health related quality of life (HRQL) of patients with chronic obstructive pulmonary disease (COPD). METHODS: A 2 year follow up study was performed in 336 patients with COPD of mean (SD) age 66 (8.2) years and mean (SD) forced expiratory volume in 1 second (FEV(1)) 33 (8)% predicted. Spirometric tests, questions regarding exacerbations of COPD, and HRQL measurements (St George's Respiratory Questionnaire (SGRQ) and SF-12 Health Survey) were conducted at 6 month intervals. RESULTS: A total of 1015 exacerbations were recorded, and 103 (30.7%) patients required at least one hospital admission during the study. After adjustment for baseline characteristics and season of assessment, frequent exacerbations had a negative effect on HRQL in patients with moderate COPD (FEV(1) 35-50% predicted); the change in SGRQ total score of moderate patients with > or =3 exacerbations was almost two points per year greater (worse) than those with <3 exacerbations during the follow up (p = 0.042). For patients with severe COPD (FEV(1) <35% predicted) exacerbations had no effect on HRQL. The change in SGRQ total score of patients admitted to hospital was almost 2 points per year greater (worse) than patients not admitted, but this effect failed to show statistical significance in any severity group. There was a significant and independent seasonal effect on HRQL since SGRQ total scores were, on average, 3 points better in measurements performed in spring/summer than in those measured in the winter (p<0.001). CONCLUSIONS: Frequent exacerbations significantly impair HRQL of patients with moderate COPD. A significant and independent effect of seasonality was also observed. PMID- 15115866 TI - Familial aggregation of FEF(25-75) and FEF(25-75)/FVC in families with severe, early onset COPD. AB - BACKGROUND: The Boston Early-Onset COPD study showed that current or ex-smoking first degree relatives of severe early onset COPD probands have significantly lower forced expiratory volume in 1 second (FEV(1)) and FEV(1)/forced vital capacity (FVC) values than current or ex-smoking control subjects, which suggests the existence of genetic risk factors for the development of COPD in response to cigarette smoking. We hypothesised that first degree relatives of early onset COPD probands may also have lower values of spirometric parameters such as forced expiratory flow at the mid-portion of forced vital capacity (FEF(25-75)) and FEF(25-75)/FVC. METHODS: Using generalised estimating equations, FEF(25-75) and FEF(25-75)/FVC were analysed in 333 first degree relatives of probands with severe early onset COPD and 83 population based controls; analyses were also performed on data stratified by smoking status. Narrow sense heritability estimates were calculated using a variance component approach. RESULTS: Significantly lower FEF(25-75) and FEF(25-75)/FVC were observed in smoking (FEF(25-75): beta -0.788 l/s (95% CI -1.118 to -0.457), FEF(25-75)/FVC: beta 20.4% (95% CI -29.3 to -11.6, p<0.0001 for both phenotypes) and non-smoking (FEF(25-75): beta -0.357 l/s (95% CI -0.673 to -0.041, p = 0.0271), FEF(25 75)/FVC: beta -9.5% (95% CI -17.1 to -1.9, p = 0.0145)) first degree relatives of early onset COPD probands. Narrow sense heritability estimates for FEF(25-75) (h(2) = 0.38) and FEF(25-75)/FVC (h(2) = 0.45) were similar to those for FEV(1) and FEV(1)/FVC. CONCLUSION: Lower values of FEF(25-75) and FEF(25-75)/FVC in non smoking first degree relatives of early onset COPD probands than in controls suggest a genetic susceptibility to develop obstructive lung disease, independent of smoking, which is magnified by exposure to deleterious environments as suggested by the further decrements in FEF(25-75) and FEF(25-75)/FVC seen in smoking first degree relatives. FEF(25-75) and FEF(25-75)/FVC have high heritability and are important intermediate phenotypes for inclusion in genetic epidemiological studies of COPD. PMID- 15115867 TI - Vasoconstrictive effects of endothelin-1, endothelin-3, and urotensin II in isolated perfused human lungs and isolated human pulmonary arteries. AB - BACKGROUND: Urotensin II (UII) has been identified as a ligand for the orphan receptor GPR14 through which it elicits potent vasoconstriction in humans and non human primates. The pulmonary vasculature is particularly sensitive; human UII (hUII) exhibits a potency 28 times that of endothelin (ET)-1 in isolated pulmonary arteries obtained from cynomolgus monkeys. However, hUII induced vasoconstriction in isolated human intralobar pulmonary arteries is variable, possibly as a result of location dependent differences in receptor density or because it is only uncovered by disease dependent endothelial dysfunction. METHODS: The vasoactivity of both hUII and gobi UII (gUII) in comparison with ET 1 and ET-3 was studied in isolated perfused lung preparations (n = 14) and isolated intralobar pulmonary arteries (n = 40, mean diameter 548 (27) microm) obtained from 17 men of mean (SE) age 67 (2) years and eight women of mean (SE) age 65 (3) years with a variety of vascular diseases. RESULTS: ET-1 (10 pM-100 nM) and ET-3 (10 pM-30 nM) elicited vasoconstriction in the lung preparations, inducing comparable increases in pulmonary arterial pressure of 24.8 (4.5) mm Hg and 14.5 (4.9) mm Hg, respectively, at 30 nM (p = 0.13). Similarly, ET-1 (10 pM 300 nM) and ET-3 (10 pM-100 nM) caused marked vasoconstriction in isolated pulmonary arteries, inducing maximal changes in tension of 4.36 (0.26) mN/mm and 1.54 (0.44) mN/mm, respectively, generating -logEC(50) values of 7.67 (0.04) M and 8.08 (0.07) M, respectively (both p<0.05). However, neither hUII nor gUII (both 10 pM-1 micro M) had any vasoactive effect in either preparation. CONCLUSION: UII does not induce vasoconstriction in isolated human pulmonary arterial or lung preparations and is therefore unlikely to be involved in the control of pulmonary vascular tone. PMID- 15115868 TI - Involvement of dendritic cells in sarcoidosis. AB - BACKGROUND: Sarcoidosis is a systemic granulomatous disease of unknown aetiology. It has been suggested that T helper type 1 (Th1) polarisation is associated with the pathophysiology of sarcoidosis, but the mechanism of skewing towards Th1 has not been elucidated. Dendritic cells (DCs) are known to regulate immune responses. This study was performed to determine whether DCs are involved in the aetiology of sarcoidosis. METHODS: The numbers of peripheral blood DCs in 24 patients with sarcoidosis were analysed and biopsy specimens from four patients were stained immunohistochemically using monoclonal antibodies. RESULTS: The numbers of both myeloid and lymphoid DC subsets were significantly decreased in the blood and mature DCs were found in the granulomas of patients with sarcoidosis. A number of interferon-gamma (IFN-gamma) producing T cells were also detected in the sarcoid granuloma, as well as many interleukin (IL)-4 producing T cells. Double staining of the biopsy specimen using anti-fascin and anti-CD3 antibodies showed an anatomical interaction between DCs and T cells. CONCLUSIONS: These findings suggest that the blood DC subsets may migrate into the affected tissues, contributing to the formation of the granulomas in sarcoidosis. It is hypothesised that the migrating DCs may regulate the T cell response in sarcoidosis, at least in the granulomatous lesions. PMID- 15115870 TI - Severe acute respiratory syndrome: report of treatment and outcome after a major outbreak. AB - BACKGROUND: The outcome is reported of a prospective uncontrolled study based on a stepwise treatment protocol during an outbreak of severe acute respiratory syndrome (SARS) in Hong Kong. METHOD: One hundred and thirty eight patients were treated with broad spectrum antibiotics, a combination of ribavirin and low dose corticosteroid, and then intravenous high dose methylprednisolone according to responses. Sustained response to treatment was defined as (1) defervescence for > or =4 consecutive days, (2) resolution of lung consolidation by >25%, and (3) oxygen independence by the fourth day without fever. Patients with defervescence who achieved either criterion 2 or 3 were classified as partial responders. Patients who fell short of criteria 2 and 3 were non-responders. RESULTS: Laboratory confirmation of SARS coronavirus infection was established in 132 (95.7%). None responded to antibiotics but 25 (18.1%) responded to ribavirin + low dose corticosteroid. Methylprednisolone was used in 107 patients, of whom 95 (88.8%) responded favourably. Evidence of haemolytic anaemia was observed in 49 (36%). A high level of C-reactive protein at presentation was the only independent predictor for use of methylprednisolone (odds ratio 2.18 per 10 mg/dl increase, 95% confidence interval 1.12 to 4.25, p = 0.02). Thirty seven patients (26.8%) required admission to the intensive care unit and 21 (15.2%) required invasive mechanical ventilation. There were 15 deaths (mortality rate 10.9%), most with significant co-morbidities, whereas 122 (88.4%) had been discharged home 4 months after the outbreak onset. CONCLUSION: The use of high dose pulse methylprednisolone during the clinical course of a SARS outbreak was associated with clinical improvement, but randomised controlled trials are needed to ascertain its efficacy in this condition. PMID- 15115872 TI - Validation of predictive rules and indices of severity for community acquired pneumonia. AB - BACKGROUND: A study was undertaken to validate the modified American Thoracic Society (ATS) rule and two British Thoracic Society (BTS) rules for the prediction of ICU admission and mortality of community acquired pneumonia and to provide a validation of these predictions on the basis of the pneumonia severity index (PSI). METHOD: Six hundred and ninety six consecutive patients (457 men (66%), mean (SD) age 67.8 (17.1) years, range 18-101) admitted to a tertiary care hospital were studied prospectively. Of these, 116 (16.7%) were admitted to the ICU. RESULTS: The modified ATS rule achieved a sensitivity of 69% (95% CI 50.7 to 77.2), specificity of 97% (95% CI 96.4 to 98.9), positive predictive value of 87% (95% CI 78.3 to 93.1), and negative predictive value of 94% (95% CI 91.8 to 95.8) in predicting admission to the ICU. The corresponding predictive indices for mortality were 94% (95% CI 82.5 to 98.7), 93% (95% CI 90.6 to 94.7), 49% (95% CI 38.2 to 59.7), and 99.5% (95% CI 98.5 to 99.9), respectively. These figures compared favourably with both the BTS rules. The BTS-CURB criteria achieved predictions of pneumonia severity and mortality comparable to the PSI. CONCLUSIONS: This study confirms the power of the modified ATS rule to predict severe pneumonia in individual patients. It may be incorporated into current guidelines for the assessment of pneumonia severity. The CURB criteria may be used as an alternative tool to PSI for the detection of low risk patients. PMID- 15115874 TI - Prognostic significance of cyclooxygenase-2 (COX-2) and expression of cell cycle inhibitors p21 and p27 in human pleural malignant mesothelioma. AB - BACKGROUND: A study was undertaken to analyse the potential prognostic value of the immunohistochemical expression of cyclooxygenase-2 (COX-2) and p27 in 29 malignant mesotheliomas already screened for the expression of p21 and p53. METHODS: Immunohistochemistry was used to determine the expression of COX-2 and p27. The correlation with survival of these factors and of p21 and p53 expression was assessed by univariate and multivariate analyses. RESULTS: A positive statistically significant correlation was found between p27 and p21 expression (p<0.0001), but there was a negative correlation between COX-2 expression and both p27 (p = 0.001) and p21 (p<0.0001). No statistically significant correlation was recorded between p53 and all the other immunohistochemical parameters. Univariate analysis showed that overall survival was strongly influenced by p21, p27, and COX-2 expression, but multivariate Cox regression analysis showed that the only immunohistochemical parameter to influence overall survival of patients with mesothelioma was COX-2. CONCLUSIONS: These findings suggest that COX-2 expression may be a useful prognostic parameter for mesothelioma. PMID- 15115876 TI - Objective assessment of criteria for selection of donor lungs suitable for transplantation. AB - BACKGROUND: Donor organ shortage severely limits lung transplantation as a therapeutic option, yet many potential donor lungs are deemed unsuitable by clinical selection criteria. METHODS: Of 39 consecutive potential donor lungs, 14 were accepted and 25 excluded by clinical selection criteria. All were evaluated prospectively by clinical assessment, bronchoscopy, and bronchoalveolar lavage (BAL) to evaluate objectively the discrimination of pulmonary infection and injury. RESULTS: Accepted donors were significantly younger than those excluded (mean (SD) age 36.7 (15.3) years v 49.5 (13.2) years; p = 0.009, unpaired t test) and were more likely to have suffered traumatic brain death (50% v 20%; p = 0.07, Fisher's exact test). Oxygenation (PaO(2):FiO(2)) was higher in accepted donors than in excluded donors (median (range) 63.2 (48-82.5) kPa v 43.1 (7.7-71.7) kPa; p = 0.0001, Mann-Whitney test). Positive formal BAL culture was more frequent in accepted donors (75%) than in those excluded (43%; p = 0.1, Fisher's exact test). There was no significant difference in the percentage and concentration of neutrophils in BAL fluid between accepted and excluded donors (median (range) 37.9 (0-96.9)% and 44.6 (0-1190)x10(3)/ml v 36 (1-98.1)% and 46 (0.2 1457)x10(3)/ml), nor in the BAL fluid concentration of tumour necrosis factor alpha (140 (0-340) pg/ml v 160 (0-760) pg/ml) or interleukin 8 (810 (33-17 600) pg/ml v 540 (0-15 110) pg/ml). CONCLUSION: Current selection criteria are poor discriminators of pulmonary injury and infection and lead to the exclusion of potentially usable donor lungs. PMID- 15115877 TI - Cough. 7: Current and future drugs for the treatment of chronic cough. AB - There are currently no effective treatments for controlling the cough response with an acceptable therapeutic ratio. However, several new mechanisms have been identified which may lead to the development of new drugs. PMID- 15115878 TI - Alpha 1-antitrypsin deficiency. 3: Clinical manifestations and natural history. AB - A review of the clinical manifestations of alpha(1)-antitrypsin (AAT) deficiency, including lung disease and liver disease, and risk factors affecting the rate of decline in lung function in AAT deficient patients. PMID- 15115879 TI - Endoglin germline mutation in a patient with hereditary haemorrhagic telangiectasia and dexfenfluramine associated pulmonary arterial hypertension. AB - Dexfenfluramine associated pulmonary arterial hypertension occurring in a patient with hereditary haemorrhagic telangiectasia related to a mutation within the endoglin gene is described. This report highlights the critical role of the TGF beta signalling pathway in this condition. PMID- 15115880 TI - Interrupter resistance. PMID- 15115881 TI - Atopic cough. PMID- 15115882 TI - Treatment of severe acute childhood asthma. PMID- 15115883 TI - Atrial septostomy in the treatment of severe pulmonary arterial hypertension. PMID- 15115884 TI - Chronic cough in adults. PMID- 15115885 TI - Pseudomembranous invasive tracheobronchial aspergillosis. PMID- 15115886 TI - Acetaminophen-induced oxidant stress and cell injury in cultured mouse hepatocytes: protection by N-acetyl cysteine. AB - The increase in cellular and mitochondrial glutathione disulfide (GSSG) levels and the GSSG:GSH ratio after acetaminophen (AAP) overdose suggest the involvement of an oxidant stress in the pathophysiology. However, the initial severe depletion of hepatocellular glutathione makes quantitative assessment of the oxidant stress difficult. Therefore, we tested the hypothesis that oxidant stress precedes the onset of cell injury in a cell culture model using 2',7' dichlorofluorescein (DCF) fluorescence as a marker for intracellular oxidant stress. Cultured primary murine hepatocytes were exposed to 5 mM AAP. DCF fluorescence, XTT reduction, lactate dehydrogenase (LDH) release, and trypan blue uptake were determined from 0 to 12 h. After glutathione depletion at 3 h, DCF fluorescence increased by 16-fold and was maintained at that level up to 12 h. At 1.5 h after AAP, a significant decrease of the cellular XTT reduction capacity was observed, which continued to decline until 9 h. Cell necrosis (LDH release, trypan blue uptake) was detectable in 20% of cells at 6 h, with a significant further increase at later time points. Pretreatment with 20 mM N-acetylcysteine (NAC) 1 h before AAP enhanced cellular glutathione content, prevented or attenuated the AAP-induced decrease of GSH levels and XTT reduction capacity, respectively, and reduced the loss of cell viability. Additionally, treatment with NAC 2 h after AAP exposure prevented further deterioration of XTT reduction at 3 h and later, and attenuated cell necrosis. Thus, AAP-induced oxidant stress precedes cell necrosis and, in cultured hepatocytes, the oxidant stress is involved in the propagation of cell injury. PMID- 15115887 TI - Characterization of atrazine biotransformation by human and murine glutathione S transferases. AB - Atrazine is one of the most widely used herbicides in the United States and has been detected, occasionally, at low levels in drinking water sources. The biotransformation of atrazine in humans has not been fully characterized. Rodent studies suggest Phase I-dominated biotransformation with minor Phase II-mediated biotransformation by glutathione S-transferase(s) (GST). In human urine, mercapturates of atrazine are significant metabolites, yet the specific GST form(s) responsible for glutathione (GSH) conjugation have not been identified. Using recombinant alpha, mu, pi and theta class human GSTs, we demonstrated that only hGSTP1-1 displays significant activity toward atrazine (7.1 nmol/min/mg protein). We also confirmed that mouse GST Pi (pi) protein is responsible for the GSH-dependent biotransformation of atrazine in mouse liver; recombinant mGSTP1-1 had a specific activity of 7.3-nmol/min/mg protein. Furthermore, cytosolic fractions from mouse and human liver conjugated atrazine with glutathione at rates of 282.3 and 3.0 pmol/min/mg, respectively. Docking studies of the atrazine GST conjugate in the hGSTP1-1 substrate-binding site were used to elucidate a basis for the dramatic difference in activity between mouse GSTP1-1 and GSTP2-2 (7.14 versus 0.02 nmol/min/mg protein, respectively). The inactivity of mGSTP2-2 appears to be attributable to an indirect structural disruption of the G-site by Pro12. Possible effects of the hGSTP1 polymorphisms were investigated. No significant differences in catalytic-specific activity were noted among purified proteins corresponding to the four hGSTP1 variants: hGSTP1(*)A (most common form), hGSTP1(*)B (Ile105Val), hGSTP1(*)C (Ile105Val, Ala114Val), and hGSTP1(*)D (Ala114Val). Overall, this work supports a physiological role for GSTs in atrazine biotransformation and indicates a novel diagnostic substrate for human and mouse GSTP1-1 proteins. PMID- 15115888 TI - PCB-induced inhibition of the vesicular monoamine transporter predicts reductions in synaptosomal dopamine content. AB - Both Aroclor mixtures and individual non-coplanar polychlorinated biphenyl (PCB) congeners reduce dopamine (DA) concentrations in cells in culture and in the brains of developing and adult laboratory animals. These reductions may involve inhibition of the dopamine transporter (DAT) and the vesicular monoamine transporter (VMAT) responsible, respectively, for the uptake of extracellular DA and the packaging of nerve terminal cytosolic DA into synaptic vesicles. However, the relative contribution of each monoamine transporter to the PCB-induced reductions in tissue DA has not been determined. Accordingly, we exposed striatal synaptosomes from adult rats to individual PCB congeners, a commercial mixture of PCBs or known monoamine transporter inhibitors; measured synaptosomal DA; and related these changes to media DA and concentrations of 3,4-dihydroxyphenylacetic (DOPAC). PCB-induced elevations in media DA concentrations are not sufficient to explain the reductions in tissue DA because known DAT inhibitors elevate media DA to a much greater extent than PCBs and yet induce similar decreases in tissue DA concentrations. On the other hand, PCB-induced elevations in DOPAC, reflective of increases in nerve terminal cytosolic DA, are sufficient to explain the reductions in tissue DA, because a known VMAT inhibitor elevates DOPAC and reduces tissue DA to an extent similar to that seen with PCBs. Taken together, these results suggest that elevations in DOPAC, reflective of increases in nerve terminal cytosolic DA due to VMAT inhibition, rather than elevations in media DA due to DAT inhibition, are largely responsible for the observed decreases in tissue DA content. PMID- 15115889 TI - Spatial activities and induction of glutamate-cysteine ligase (GCL) in the postimplantation rat embryo and visceral yolk sac. AB - Glutathione (GSH) synthesis is differentially regulated in the embryo and visceral yolk sac (VYS) of the developing rat conceptus. The innate capacity to respond to environmental insult and chemical exposure by inducing de novo GSH synthesis may help to determine overall cell sensitivity and/or resistance to chemically induced malformation. Specific activities of glutamate-cysteine ligase (GCL), the rate limiting enzyme in GSH synthesis, were determined by measuring the formation of gamma-glutamylcysteine (GC) in homogenates prepared from rat embryos and VYSs. GC formation increased linearly with time and with relative protein concentration. Specific activities were found to be 60.5 +/- 3.2 and 118.9 +/- 4.2 pmol GC/mg protein/min in the gestational day (GD) 10 embryo and VYS, respectively, and 22.7 +/- 0.4 and 71.3 +/- 0.6 pmoles GC/mg protein/min in the respective GD 11 embryo and VYS. Apparent kinetic constants determined from embryo and VYS homogenates gave respective apparent K(m) values for glutamate of 0.75 and 1.38 mM and for cysteine 0.03 mM in both tissues. Apparent V(max) values were higher in the VYS in each case, corresponding with a lower apparent K(m) and higher GCL activity. GCL specific activities increased significantly following a 24 h in vitro exposure to diethyl maleate (DEM) and diamide, but remained unchanged following exposure to prostaglandin A(2) (PGA(2)) and t-butylated hydroxytoluene (BHT). Basal expression of GCL catalytic subunit (GCL(C)) and regulatory subunit (GCL(R)) was 59- and 25-fold higher in VYS, respectively, compared to the embryo. Quantitative real-time fluorescence reverse transcriptase polymerase chain reaction (RT-PCR) showed that following DEM and diamide treatment, GCL(C) expression increased up to 19-fold in embryonic tissues but was not induced in the VYS. Only DEM increased the expression of the light/regulatory subunit GCL(R) in the embryo (8-fold). Densitometry of immunoblots revealed approximately 75% more GCL(C) in the VYS than in the embryo. Following treatments, a marked increase was induced in embryonic GCL(C) content with both DEM (85%) and diamide (19%), but in the VYS, only DEM caused an increase in GCL(C) protein (38%). PMID- 15115890 TI - Re: Viberg H, et al. Neurobehavioral derangements in adult mice receiving decabrominated diphenyl ether (PBDE 209) during a defined period of neonatal brain development. Toxicol Sci. 2003 Nov;76(1):112-20. PMID- 15115892 TI - What happens? PMID- 15115893 TI - JCO interviews Richard P. McLaughlin, DDS, on facial and dental planning. Interview by Robert G. Keim. PMID- 15115894 TI - The Readers' Corner. 1. Do you use non-compliance appliances? PMID- 15115895 TI - JCO survey of referring dentists. PMID- 15115896 TI - A new Class II distalizer. PMID- 15115897 TI - Class II correction with the Twin Force Bite Corrector. PMID- 15115898 TI - A modified monobloc for treatment of young children with obstructive sleep apnea. PMID- 15115899 TI - Nicorandil protects against lethal ischemic ventricular arrhythmias and up regulates endothelial nitric oxide synthase expression and sulfonylurea receptor 2 mRNA in conscious rats with acute myocardial infarction. AB - Nicorandil is an adenosine triphosphate sensitive K (K-ATP) channel opener and a nitric oxide donor. K-ATP channels and nitric oxide are important factors in ischemic preconditioning, which in turn suppresses reperfusion arrhythmias. The present study sought to evaluate whether nicorandil suppresses ischemic-induced ventricular arrhythmias and enhances sulfonylurea receptors (SUR 2; subunit of K ATP channel), endothelial nitric oxide (eNOS), and inducible nitric oxide (iNOS) expression in the left ventricle after myocardial infarction without reperfusion. Thirty male Sprague-Dawley rats at 7 weeks of age were separated into three groups, as follows. Acute myocardial infarction was induced in twenty rats by ligating the left main coronary artery. Ten of these twenty rats were continuously administered nicorandil at 3 mg/kg/day i.p. The other ten rats were left untreated. The ten controls were untreated and sham-operated. After coronary ligation, ventricular arrhythmias were evaluated from stored ECG signals. At 24 hours after treatment, eNOS, iNOS, and SUR2 mRNA levels and eNOS, iNOS expression in the left ventricle were determined by reverse transcription polymerase chain reaction (RT-PCR) and by immunohistochemical staining, respectively. Nicorandil suppressed the total number of ventricular arrhythmias from 1 to 2 hours, the total duration of ventricular tachycardia from 2 to 3 hours, and that of ventricular fibrillation from 1 to 2 and from 4 to 5 hours after coronary ligation. Nicorandil improved the survival rate 24 hours after coronary ligation. Levels of SUR2 mRNA increased only in left ventricles treated with nicorandil, particularly in the non-ischemic myocardium. eNOS mRNA was enhanced 2.2-fold in the area at risk in infarcted controls compared to sham-operated rats. In the non ischemic area and area at risk of rats treated with nicorandil compared to sham operated rats, eNOS mRNA was enhanced 3.3- and 2.7-fold, respectively. Staining indicated that the highest concentrations of eNOS occurred in the endothelium and myocardium of the non-ischemic area of rats treated with nicorandil. iNOS mRNA was present in both the area at risk and the non-ischemic area only in infarcted rats, and levels thereof were higher in the area at risk than in the non-ischemic area. However, there was no difference in iNOS mRNA levels between nicorandil treated rats and controls. iNOS exhibited stronger staining in the area at risk than in the non-ischemic area of both nicorandil-treated and infarcted controls, with no differences between these two groups of rats. The mechanisms of protection against lethal ventricular tachyarrhythmia in nicorandil may increase nitric oxide release by upregulated eNOS expression through the opening of K-ATP channels and/or a K-ATP channels opener itself after acute myocardial infarction. PMID- 15115900 TI - Effect of the cardioselective, sarcolemmal K(ATP) channel blocker HMR 1098 on atrial electrical remodeling during pacing-induced atrial fibrillation in dogs. AB - PURPOSE: The progressive shortening of the atrial effective refractory period (ERP) during atrial fibrillation (AF) might be due to the activation of the KATP channels by rapid atrial rates. We tested the hypothesis that the cardioselective, sarcolemmal KATP channel blocker HMR 1098 would prevent atrial ERP shortening during AF. METHODS AND RESULTS: Nine dogs were treated with HMR 1098 (3 mg/kg bolus injection followed by a continuous intravenous (i.v.) infusion at 17 microg/kg/min rate maintained throughout the study) and 7 dogs served as controls receiving i.v. saline. Pharmacological autonomic blockade was induced by i.v. administration of atropine (0.04 mg/kg) and propranolol (0.2 mg/kg) and maintained throughout the study by continuous i.v. infusion of atropine (0.007 mg/kg/h) and propranolol (0.04 mg/kg/h). Rapid right atrial pacing at 50 msec cycle length (CL) was initiated and maintained for 6 hours. High right atrial ERP (HRA-ERP) and corrected sinus node recovery time (HRA cSNRT), coronary sinus ERP (CS-ERP) and corrected SNRT (CS-cSNRT) at three (400, 300, 200 msec) CLs were measured before and after pacing at different time points. Baseline values were not different between control and treated dogs. In the control group the HRA-ERP progressively shortened (from 179 +/- 21 msec at baseline to 161 +/- 23 msec at 360 min at 400 msec CL) ( p = 0.002), with a gradual decrease, loss or inversion of ERP rate adaptation at shorter (300, 200 msec) CLs. HMR 1098 treatment did not prevent the shortening of HRA-ERP during the first 2 to 3 hours of rapid atrial pacing. However, beginning at 180-240 min, HMR 1098 increased the HRA-ERP ( p = 0.01) to baseline by 360 min. HMR 1098 treatment did not prevent another feature of atrial electrical remodeling, the flattening or inversion of ERP rate adaptation. In neither group did CS-ERP shortening occur. The maximum cSNRT at 360 min prolonged significantly in both groups during HRA and CS pacing as well compared with baseline. CONCLUSIONS: HMR 1098 treatment did not prevent the shortening of HRA-ERP, the salient feature of atrial electrical remodeling in the first 2 to 3 hours of rapid atrial rates, but did prevent it thereafter. Another characteristic feature of atrial electrical remodeling, the flattening or inversion of physiological ERP rate adaptation was not prevented by HMR 1098 treatment. Sinus node depression was detectable after short-term (6 hours) rapid atrial pacing and was not affected by HMR 1098. PMID- 15115901 TI - 17ss oestradiol does not attenuate the response to angiotensin I and II during long term therapy in postmenopausal women. AB - PURPOSE: It is thought that oestrogen replacement therapy may reduce the conversion of angiotensin I to angiotensin II in postmenopausal women by inhibiting angiotensin converting enzyme. We sought to determine the effects of 3 months oestrogen replacement therapy (ERT) on the response to angiotensin I and II in postmenopausal women. METHODS: Eighteen postmenopausal women were randomised to either three months of 2 mg oral oestradiol or placebo in a double blind, placebo controlled, protocol. Change in forearm blood flow (FBF), in response to brachial arterial infusion of increasing concentrations of angiotensin I and angiotensin II was measured, pre-randomisation, after 1 months randomised therapy and after 3 months therapy, using venous occlusion plethysmography. RESULTS: Oestrogen treatment had no effect on baseline FBF. The mean (SD) peak reductions in FBF with 64 pmol/min angiotensin I pre randomisation, after 1 month and after 3 months treatment with placebo were 62(11), 65(15) and 57(8)%. The corresponding reductions with the peak dose of angiotensin II (16 pmol/min) were 53(8), 48(12) and 47(11)%. In the oestradiol group, the peak responses to angiotensin I were 56(8), 53(11) and 44(29)% pre randomisation, after 1 months treatment and after 3 months treatment. The corresponding reductions in response to angiotensin II were 37(15), 47(15) and 45(19)%. Oestradiol did not affect the response to either angiotensin I or angiotensin II. CONCLUSION: This study does not support the idea that ERT might exert a cardioprotective effect through inhibition of the renin-angiotensin system. PMID- 15115902 TI - Lack of anti-ischemic efficacy of the potassium channel opener bimakalim in patients with stable angina pectoris. AB - Aim of the study was to evaluate anti-ischemic, hemodynamic and neurohumoral effects of bimakalim, a novel selective K(+)-channel opener, in patients with stable angina pectoris and reproducible ST-segment depression. METHODS AND RESULTS: 86 patients with angiographic signs of CAD were involved in two randomised, placebo-controlled, double-blind trials with single high (0.1, 0.3 or 0.6 mg) and low (0.025 or 0.05 mg) doses of oral bimakalim. The anti-anginal efficacy was evaluated by analysis of ST-segment depression within exercise or RV pacing. A parallel assessment of hemodynamic parameters was done by means of right-heart catheterization. Given in high doses, bimakalim acted as a potent vasodilator and decreased SBP by 15 mm Hg. This was associated with a reflex activation of sympathetic nervous system resulting in an increase in heart rate by 25 /min, a 14% rise of myocardial oxygen consumption and a 63% elevation of noradrenaline plasma level. Low doses of bimakalim had no significant effect on hemodynamics and oxygen consumption. In exercise-induced angina pectoris, administration of bimakalim was neither associated with attenuation of ST-segment depression nor resulted in prolongation of time to 0.1 mV ST-segment depression. CONCLUSION: The results of the present study suggest that bimakalim has a dose dependent vasodilatatory activity but exerts no anti-ischemic benefits in patients with exercise-induced angina pectoris due to coronary artery disease. PMID- 15115903 TI - Anti-ischaemic response to sublingual nitroglycerin during oral administration of isosorbide dinitrate in patients with stable angina pectoris: when does cross tolerance occur? AB - The purpose of this study was to evaluate the efficacy of sublingual nitroglycerin (NTG) during treatment with oral sustained-release isosorbide dinitrate (ISDN) in two doses: 80 mg and 120 mg. In a double-blind crossover design study 38 men with stable angina initially received either an oral placebo (OP) or ISDN. All patients received either NTG 0.5 mg or sublingual placebo (SLP) 2.5 h after OP ingestion, but only NTG 2.5 h after ISDN. The same pattern was used in the first ingestion and in long-term OP or ISDN therapy for 7 days (OP and ISDN every 6 h, and ISDN once daily). The efficacy of NTG was evaluated by analyzing walking time to ischaemia (WTI) during exercise tests performed 5 minutes after NTG or SLP administration, and the efficacy of ISDN 2 h and 6 h after oral ingestion. In the first ingestion NTG significantly improved WTI ( p < 0.0001) by 42.7% after OP, by 46.5% after 80 mg ISDN and by 52.1% after 120 mg. After long-term OP therapy NTG prolonged WTI ( p < 0.01) by 15.6%, during once daily ISDN treatment, an 80 mg dose prolonged WTI by 22.8% and a dose of 120 mg by 36.5%. However, NTG did not improve WTI in q.i.d. therapy. Six hours after the first 80 mg ISDN ingestion WTI improved ( p < 0.0001) by 66.0%, and after 120 mg by 58.4%. Following once-daily therapy ISDN prolonged WTI ( p < 0.0001) by 27.2% after an 80 mg dose and by 36.2% after a dose of 120 mg. No improvement was observed in q.i.d. treatment. Thus, severe tolerance to ISDN abolishes the anti ischaemic effects of NTG, and appropriate regimens of ISDN have considerable anti anginal effects during chronic administration. PMID- 15115904 TI - The benefits of early combination treatment of carvedilol and an ACE-inhibitor in mild heart failure and left ventricular systolic dysfunction. The carvedilol and ACE-inhibitor remodelling mild heart failure evaluation trial (CARMEN). AB - AIMS: Heart failure (HF) treatment guidelines of the ESC recommend ACE-inhibitors (ACE-I) as first-line treatment and beta-blockers added if patients remain symptomatic. CARMEN explored the need for combined treatment for remodelling and order of introduction by comparing the ACE-I enalapril against carvedilol and their combination. METHODS: In a parallel-group, 3-arm study of 18 months duration, 572 mild heart failure patients were randomly assigned to carvedilol (N = 191), enalapril (N = 190) or their combination (N = 191). In the latter, carvedilol was up-titrated before enalapril. Left ventricular (LV) remodelling was assessed by transthoracic echocardiography (biplane, modified Simpson) at baseline and after 6, 12 and 18 months of maintenance therapy. Primary comparisons considered the change in LV end-systolic volume index (LVESVI) from baseline to month 18 between the combination and enalapril, and between carvedilol and enalapril. RESULTS: In the first primary comparison, LVESVI was reduced by 5.4 ml/m2 (p = 0.0015) in favour of combination therapy compared to enalapril. The second primary comparison tended to favour carvedilol to enalapril (NS). In the within treatment arm analyses, carvedilol significantly reduced LVESVI by 2.8 ml/m2 (p = 0.018) compared to baseline, whereas enalapril did not. LVESVI decreased by 6.3 ml/m2 (p = 0.0001) with combination therapy. All three arms showed similar safety profiles and withdrawal rates. CONCLUSION: CARMEN is the first study to demonstrate that early combination of ACE-I and carvedilol reverses LV remodelling in patients with mild to moderate HF and LV systolic dysfunction. The results of the CARMEN study support a therapeutic strategy in which the institution of beta-blockade should not be delayed. PMID- 15115905 TI - Fluvastatin reduces cardiac mortality in patients with coronary heart disease. AB - PURPOSE: To test the effectiveness of fluvastatin, 40-80 mg, in reducing the occurrence of cardiac and all-cause mortality in patients with coronary heart disease (CHD). METHODS: Meta-analysis of all clinical trials that assessed the effects of fluvastatin in CHD patients on major adverse cardiac events (MACE) as a prespecified endpoint was performed. A pooled analysis of four studies (n = 3525) was performed on an intent-to-treat basis. Clinical endpoints were the incidence, and time to first occurrence, of MACE (cardiac death, nonfatal MI, revascularization), noncardiac death, or all-cause death. Lipid parameters were also analyzed. RESULTS: Fluvastatin treatment significantly prolonged the time to cardiac death (p = 0.0174) and the time to cardiac death or nonfatal MI (p = 0.0055) compared with placebo. Fluvastatin significantly reduced the risk of any MACE (Cox risk ratio [RR], 0.85; 95% confidence interval [CI], 0.73-0.98), cardiac death (RR, 0.53; 95% CI, 0.31-0.90), cardiac death or MI (RR, 0.66; 95% CI, 0.49-0.89), all-cause death (RR, 0.65; 95% CI, 0.45-0.94) and all-cause death or MI (RR, 0.69; 95% CI, 0.53-0.90). Fluvastatin significantly lowered total cholesterol and low-density lipoprotein cholesterol levels and was well tolerated, with no cases of rhabdomyolysis in any of the studies assessed in the meta-analysis. CONCLUSIONS: This meta-analysis demonstrates clear beneficial effects of fluvastatin on cardiac and all-cause mortality in CHD patients, and supports the use of fluvastatin to reduce the incidence of MACE in a wide range of at-risk patients. PMID- 15115906 TI - Hemorrhagic transformation of ischemic stroke associated with enoxaparin and aspirin concomitant therapy. PMID- 15115908 TI - Differences among the cell wall galactomannans from Aspergillus wentii and Chaetosartorya chrysella and that of Aspergillus fumigatus. AB - The alkali extractable and water-soluble cell wall polysaccharides F1SS from Aspergillus wentii and Chaetosartorya chrysella have been studied by methylation analysis, 1D- and 2D-NMR, and MALDI-TOF analysis. Their structures are almost identical, corresponding to the following repeating unit: [--> 3)-beta-D-Gal f (1 --> 5)-beta-D-Gal f-(1 -->]n --> mannan core. The structure of this galactofuranose side chain differs from that found in the pathogenic fungus Aspergillus fumigatus, in other Aspergillii and members of Trichocomaceae: [--> 5)-beta-D-Gal f-(1 -->]n --> mannan core. The mannan cores have also been investigated, and are constituted by a (1 --> 6)-alpha-mannan backbone, substituted at positions 2 by chains from 1 to 7 residues of (1 --> 2) linked alpha-mannopyranoses. PMID- 15115907 TI - Tumor galectinology: insights into the complex network of a family of endogenous lectins. AB - Beta-Galactosides of cell surface glycoconjugates are docking sites for endogenous lectins of the galectin family. In cancer cells, primarily galectins-1 and -3 have been studied to date. With the emergence of insights into their role in growth control, resistance to or induction of apoptosis and invasive behavior the notion is supported that they can be considered as functional tumor markers. In principle, the same might hold true for the other members of the galectin family. But their expression in tumors has hitherto been a subject of attention only to a very limited extent. Pursuing our concept to define the complexity of the galectin network in cancer cells and the degree of functional overlap/divergence with diagnostic/therapeutic implications, we have introduced comprehensive RT-PCR monitoring to map their galectin gene expression. The data on so far less appreciated galectins in this context such as galectins-4 and -8 vindicate this approach. They, too, attach value to extend the immunohistochemical panel accordingly. Our initial histopathological and cell biological studies, for example on colon cancer progression, prove the merit of this procedure. Aside from the detection of gene expression profiles by RT-PCR, the detailed molecular biological monitoring yielded further important information. We describe different levels of regulation of galectin production in colon cancer cells in the cases of the tandem-repeat-type galectins-8 and -9. Isoforms for them are present with insertions into the peptide linker sequence attributed to alternative splicing. Furthermore, variants with distinct amino acid substitutions (galectin-8, Po66-CBP, PCTA-1, CocaI/II and galectin 9/ecalectin) and generation of multiple mRNA species, notably those coding for truncated galectin-8 and -9 versions with only one lectin site, justify to portray these two family members not as distinct individuals but as groups. In aggregate, the ongoing work to thoroughly chart the galectin network and to disentangle the individual functional contributions is expected to make its mark on our understanding of the malignant phenotype in certain tumor types. PMID- 15115909 TI - Galectin-4 in normal tissues and cancer. AB - Galectin-4 belongs to a subfamily of galectins composed of two carbohydrate recognition domains within the same peptide chain. The two domains have all the conserved galectin signature amino acids, but their overall sequences are only approximately 40% identical. Both domains bind lactose with a similar affinity as other galectins, but their respective preferences for other disaccharides, and larger saccharides, are distinctly different. Thus galectin-4 has a property of a natural cross-linker, but in a modified sense since each domain prefers a different subset of ligands. Similarly to other galectins, galectin-4 is synthesized as a cytosolic protein, but can be externalized. During development and in adult normal tissues, galectin-4 is expressed only in the alimentary tract, from the tongue to the large intestine. It is often found in relatively insoluble complexes, as a component of either adherens junctions or lipid rafts in the microvillus membrane, and it has been proposed to stabilize these structures. Strong expression of galectin-4 can be induced, however, in cancers from other tissues including breast and liver. Within a collection of human epithelial cancer cell lines, galectin-4 is overexpressed and soluble in those forming highly differentiated polarized monolayers, but absent in less differentiated ones. In cultured cells, intracellular galectin-4 may promote resistance to nutrient starvation, whereas--as an extracellular protein--it can mediate cell adhesion. Because of its distinct induction in breast and other cancers, it may be a valuable diagnostic marker and target for the development of inhibitory carbohydrate-based drugs. PMID- 15115910 TI - Molecular cloning of goat Mannose 6-phosphate receptors, MPR 300 and 46. AB - Mannose 6-phosphate receptor proteins (MPR 300 and 46) are type 1 transmembrane glycoproteins that mediate transport of lysosomal enzymes to lysosomes. In a recent study we have purified both receptors from goat liver and raised antibodies. An ELISA method was developed that allowed quantification of both receptors in different tissues of goat and chicken and an immuno-affinity method was also developed to purify the receptors. In the present study to understand the structural similarities of the goat receptors to other known receptor proteins, we have prepared cDNA clones for both receptors by RT-PCR approach. A partial cDNA clone (1.368 kb) for the MPR 300 protein, and a full length cDNA clone (0.84 kb) for the MPR 46 protein were obtained. MPR 300 exhibits typical conserved cassette structure in the amino terminal domain similar to other known vertebrate MPR proteins with the conserved cysteine residues and the ligand binding arginine residue in the third domain. MPR 46 exhibits high degree of sequence homology to other known MPR proteins with the conserved cysteine residues, the transmembrane domain and the cytoplasmic tail. mRNA transcript size for both receptors were comparable with that of other vertebrates. PMID- 15115912 TI - Investigation of the association between 5-HT3A receptor gene polymorphisms and efficiency of antiemetic treatment with 5-HT3 receptor antagonists. AB - OBJECTIVES: Acute cytostatic drug induced nausea and vomiting is provoked by a release of endogenous serotonin that mediates its effect by binding to the 5 hydroxytryptamine type 3 (5-HT3) receptors. The most effective antiemetic drugs are the 5-HT3 receptor antagonists. Nevertheless about 30% of the patients do not respond satisfactorily. Five 5-HT3 receptor genes (5-HT(3A-E)) with high sequence homology have been identified. Two subunits, the 5-HT3A and 5-HT3B are expressed in anatomical structures known to be involved in the mechanism of acute cytostatic drug induced emesis. METHODS: We included 242 cancer patients at their first day of chemotherapy to investigate the influence of genetic polymorphisms of the 5-HT3A receptor gene on the intensity of nausea and vomiting which was documented using standardized interviews and visual analog scales. RESULTS: Sequencing of the entire 5-HT3A receptor gene of all patients revealed 21 polymorphisms, two of them were amino acid substitutions (Ala33Thr, Met257Ile). Linkage disequilibrium analysis revealed that 15 polymorphisms of the 5-HT3A receptor gene are partially linked to each other. However, none of the haplotypes was significantly associated with the intensity of cytostatic induced nausea and vomiting. CONCLUSION: Polymorphisms and haplotype analysis of the 5-HT3A receptor gene may not serve as a pharmacogenetic predictor of the antiemetic treatment with 5-HT3 receptor antagonists in cancer patients. PMID- 15115911 TI - Developmental changes in the biochemical and immunological characters of the carbohydrate moiety of neuroglycan C, a brain-specific chondroitin sulfate proteoglycan. AB - Neuroglycan C (NGC), a brain-specific transmembrane proteoglycan, is thought to bear not only chondroitin sulfate but also N- and O-linked oligosaccharides on its core protein. In this study, we isolated and purified NGC from rat brains at various developmental stages by immunoaffinity column chromatography or by immunoprecipitation, and examined the structural characters of its carbohydrate moiety. The chondroitin sulfate disaccharide composition of NGC at postnatal day 10 was significantly different from those of two secreted chondroitin sulfate proteoglycans, neurocan and phosphacan, purified from the brain at the same developmental stage; higher levels of 4-sulfate unit and E unit, a disulfated disaccharide unit, and a lower level of 6-sulfate unit. The levels of both 6 sulfate and E units decreased with a compensatory increase of 4-sulfate unit with postnatal development of the brain. Lectin-blot analysis of the NGC core glycoprotein prepared by chondroitinase digestion confirmed that NGC actually bore both N- and O-linked carbohydrates, and also revealed that lectin-species reactive with NGC did not always recognize other brain-specific proteoglycans, neurocan and phosphacan, and vice versa, even though they were isolated from the brain at the same stage. The reactivity of NGC with lectins and with the HNK-1 antibody markedly changed as the brain matured. These findings indicate that the structure of the carbohydrate moiety of NGC is developmentally regulated, and differs from those of neurocan and phosphacan. The developmentally-regulated structural change of the carbohydrates on NGC may be partly implicated in the modulation of neuronal cell recognition during brain development. PMID- 15115913 TI - Impact of CYP2D6 intermediate metabolizer alleles on single-dose desipramine pharmacokinetics. AB - This study utilized cytochrome P450 2D6 (CYP2D6) genotypes to explain variability of desipramine pharmacokinetics in a cohort of non-poor metabolizer individuals. In an interaction study utilizing desipramine as a probe, genotyping for the CYP2D6*3, *4, *5 and *6 alleles was used to screen out CYP2D6 poor metabolizers. Individuals were categorized according to these and additional alleles (CYP2D6*2, *9, *10, *17, *41 and x2). Genotypes of individuals heterozygous for two or three of *2, *17 and *41 alleles were confirmed by molecular haplotyping. Pharmacokinetic parameters of desipramine were analysed according to CYP2D6 category. Molecular haplotyping was necessary to definitively categorize four of 16 individuals. A subject who had unusually high plasma elimination half-time, exposure and metabolic ratios carried an intermediate metabolizer (IM) *9 allele in combination with a non-functional allele. This combination has a population frequency of less than 1 : 200. Individuals with *1/*1, *1/*2 and *2/*2 genotypes had lower than average plasma elimination half-time, exposure and metabolic ratios. For desipramine, additional genotyping of CYP2D6 IM alleles helped define subgroups of the CYP2D6-positive cohort. This suggests that genotyping for IM alleles will aid in interpretation of clinical trials involving CYP2D6 substrates. Due to the diversity of IM alleles, molecular haplotyping may be necessary to fully characterize CYP2D6 genotype-phenotype relationships. PMID- 15115914 TI - Human controlled ovarian hyperstimulation outcome is a polygenic trait. AB - This study aimed to evaluate the association between follicle-stimulating hormone (FSH) hormone efficacy and FSHR, CYP19, ESR1 and ESR2 genes using single nucleotide polymorphism analyses. One hundred and seventy women with conserved ovarian function undergoing controlled ovarian stimulation (COS) with daily exogenous recombinant FSH administration. Women were categorized as poor responders to FSH (three or less ovarian follicles observed at the end of cycle) or normal responders (more than three follicles). The outcome is the number of normal/poor responders as defined by the number of follicles obtained during COS. The DNA markers studied are located in genes related to the FSH mechanism of action (FSH receptor, CYP19 aromatase and oestrogen receptors alpha and beta genes). We conducted an association study between the COS outcome and selected DNA markers using two-point and multi-locus genetic association studies. Genotype pattern tracking in extreme phenotypes and multi-locus analysis using Sumstat and PM algorithms provided significant evidences of genetic interaction between FSHR, ESR1 and ESR2 markers in relation to COS outcome (P = 0.0015). Our results support the hypothesis that a discrete set of genes, related to the FSH hormone mechanism of action, controls the ovarian response to FSH in humans. An oligogenic model including specific FSHR, ESR1 and ESR2 genotype patterns may partially explain the poor response to FSH hormone during controlled ovarian stimulation treatments. The existence of genetic heterogeneity is also suspected. PMID- 15115915 TI - Glutathione-S-transferase M1, M3, P1 and T1 polymorphisms and severity of lung disease in children with cystic fibrosis. AB - OBJECTIVES: Progression and severity of lung disease differs markedly and early between patients with cystic fibrosis (CF). We investigated the hypothesis that polymorphisms in the detoxifying enzymes glutathione-S-transferase (GST) could influence phenotypic presentation of lung disease in CF. METHODS: Genotypes for GSTM1, GSTM3, GSTP1 and GSTT1 were determined in a cohort of 146 children with CF by PCR-based methods. Pulmonary function, assessed by spirometric measures of forced expiratory volume in one second (FEV1) and forced vital capacity (FVC), was analysed in children at the age of 9. RESULTS: No association between spirometric measurements, and GSTM1, GSTP1 or GSTT1 genotypes was found. As compared with patients homozygous for GSTM3*A allele, CF children carrying the GSTM3*B allele displayed a significant better lung function, assessed by both mean values of FEV1 and of FVC (respectively P = 0.01 and P = 0.002). These correlations remained significant after adjustment for potential confounding factors (respectively adjusted P = 0.008 and P = 0.002) and also in subgroups of CF patients who carry the deltaF508 CFTR mutation. Haplotype analysis of GSTM3 in combination with GSTM1 indicated that the positive impact of GSTM3*B allele on pulmonary performances was barely influenced by the GSTM1 genotypes of CF children. CONCLUSIONS: These data provide the first evidence suggesting that polymorphism of the GSTM3 gene contributes to clinical severity in CF, which may have prognostic significance and could prompt to start a more targeted therapy in young patients with CF. PMID- 15115916 TI - Interaction between NOTCH4 and catechol-O-methyltransferase genotypes in schizophrenia patients with poor response to typical neuroleptics. AB - OBJECTIVE: In this study we attempted to show that the interaction between NOTCH4 and catechol-O-methyltransferase (COMT) polymorphism predicts the response to typical neuroleptics in schizophrenia. Our sample consisted of 94 Finnish patients with DSM-IV schizophrenia and 98 controls. METHODS: Several studies have connected COMT and NOTCH4 genes to schizophrenia. We have previously shown that COMT polymorphism is significantly associated with treatment response in schizophrenia. NOTCH4 SNP2 polymorphism has been associated with age of onset in schizophrenia, but there is also a trend that this polymorphism may predict response to typical neuroleptics. In the present sample, there is a strong gene gene interaction between these genes (P = 0.003) and they have additive effect in treatment response. RESULTS: Patients carrying both NOTCH4 C/C genotype and COMT low/low genotype, had more than ten times higher risk of being a non-responder than responder to treatment with typical neuroleptics [OR = 10.25 (95% CI 2.21 47.53), P < 0.001]. This combination of genotypes is also more common in patients considered non-responders than in controls [OR = 3.00 (95% CI 1.33-6.76), P = 0.007]. CONCLUSION: Our results suggest that an interaction between COMT and NOTCH4 genotypes may predict the treatment response to typical neuroleptics in patients with schizophrenia. PMID- 15115917 TI - Variable expression of P-glycoprotein in the human placenta and its association with mutations of the multidrug resistance 1 gene (MDR1, ABCB1). AB - The MDR1 gene product P-glycoprotein in the human placenta is important for protecting the fetus from unintended, harmful drug exposure, but also for limiting the access of therapeutic drugs to the fetus after maternal drug intake. A polymorphism in exon 26 of the MDR1 gene (C3435T) has previously been shown to be associated with reduced P-glycoprotein expression in the small intestine, kidney and lymphocytes. In the present study, we examined systematically whether MDR1 polymorphisms also have an impact on P-glycoprotein expression in the human placenta. MDR1 mRNA and P-glycoprotein were analysed in 73 full-term human placentas of Caucasians, as well as respective MDR1 genotypes/haplotypes, for the C3435T and G2677T/A polymorphisms of mothers and infants. MDR1 mRNA levels were not different between these genotype groups. However, P-glycoprotein expression was significantly lower when both mother and infant were homozygous for the 3435T allele (TT/tt) compared to maternal and fetal homozygotes for the C-allele (0.40 +/- 0.18 a.u. for TT/tt versus 0.66 +/- 0.30 a.u. for CC/cc, P = 0.01). Moreover, placentas from mothers carrying both polymorphisms (3435T and 2677T; TT/TT) also had a significantly lower P-glycoprotein expression (0.31 +/- 0.12 a.u.) compared to placentas of wild-type individuals (CC/GG, 0.71 +/- 0.31 a.u., P = 0.02). Taken together, the MDR1 polymorphisms C3435T and G2677T are associated with altered P-glycoprotein expression in the human placenta, and may have clinical consequences due to genetically determined, variable drug exposure of the fetus. PMID- 15115918 TI - A 6 bp polymorphism in the thymidylate synthase gene causes message instability and is associated with decreased intratumoral TS mRNA levels. AB - OBJECTIVE: A 6 bp deletion polymorphism in the thymidylate synthase (TS) gene was investigated in order to determine its function. METHODS: A luciferase system was used to investigate the function of the 6 bp/1494 polymorphism in vitro. A group of 43 patients with colorectal carcinoma were evaluated for the 6 bp/1494 polymorphism and for intratumoral TS mRNA levels in vivo. RESULTS: The 3'UTR of TS containing the +6 bp polymorphism resulted in an approximate 35% decrease in luciferase activity and mRNA levels, while the TS-3'UTR bearing the -6 bp deletion resulted in an approximate 70% decrease in luciferase activity and mRNA levels. The TS-3'UTR construct containing the -6 bp/1494 deletion also had a higher rate of message degradation compared to the +6 bp/1494 construct. Individuals homozygous for the insertion (+6 bp/+6 bp) had significantly higher TS mRNA levels compared to individuals that were homozygous for the deletion (-6 bp/-6 bp) (P < 0.007). We determined the frequency of the -6 bp/1494 deletion polymorphism to be 41% in non-Hispanic whites, 26% in Hispanic whites, 52% in African-Americans and 76% in Singapore Chinese. CONCLUSIONS: These results suggest that the -6 bp/1494 deletion polymorphism in the 3'UTR of TS is associated with decreased mRNA stability in vitro and lower intratumoral TS expression in vivo. Further, the 6 bp/1494 polymorphism varies greatly within different ethnic populations and is in linkage disequilibrium with the TS 5' tandem repeat enhancer polymorphism. Taken together, these data suggest that the 6 bp/1494 polymorphism may be a useful screening tool in predicting TS mRNA expression. PMID- 15115919 TI - A novel polymorphism in the promoter region of human UGT1A9 gene (UGT1A9*22) and its effects on the transcriptional activity. AB - The human UDP-glucuronosyltransferase, UGT1A9, catalyses glucuronidations of various endobiotics and xenobiotics. In the present study, all exons, exon-intron junctions, and the 5'-flanking region (-273 bp) of the UGT1A9 gene in a Japanese subject were sequenced. One base insertion of thymidine in a promoter region of the UGT1A9 gene resulting in A(T)10AT was identified compared to the reference sequence of AF297093 (A(T)9AT). The allele was termed UGT1A9*22. A polymerase chain reaction-single strand conformation polymorphism method was developed to genotype the allele. The allele frequencies of the mutation in 87 Japanese, 50 Caucasian and 50 African-American subjects were 60%, 39% and 44%, respectively. The significance of the polymorphism was investigated by the construction of luciferase reporter plasmids containing 170 bp of the 5'-flanking region of the gene transfected into human hepatoma HepG2 cells. The luciferase activity of the promoter construct containing the A(T)10AT sequence was 2.6-fold higher than that of the construct containing the A(T)9AT sequence. In conclusion, the mutant allele with one base insertion in the promoter region of the UGT1A9 gene would alter the level of enzyme expression and the metabolism of those drugs that are substrates of UGT1A9. PMID- 15115920 TI - Colitis: problems in pathologic and clinical diagnosis: indeterminate, microscopic, and diverticulitis. Yale University Workshop Report. October, 2003. Norwalk, Connecticut, USA. PMID- 15115921 TI - The natural history of diverticulitis: fact and theory. AB - Epidemiological and anatomic evidence indicates that approximately 60% of humans of westernized societies living into the sixth decade will develop diverticulosis of the colon. The cause remains unknown, but epidemiological studies indicate it is a combination of decreased dietary fiber intake and increased intracolonic pressure. The intraluminal pressure exerted on the wall causes a diverticular outpocketing at any one of the three areas in which vessels enter the wall. In this paper, we advance a hypothesis that fiber deficiency not only leads to diverticula formation but also causes a change in the microecology that results in decreased colon immune response and permits a low-grade chronic inflammatory process that precedes a full-blown acute diverticulitis. Pathophysiologic studies reveal that complications do not occur until there is microperforation through the wall of the diverticulum into the pericolic tissue. The perforation might be small and cause a microabscess, or extend to a phlegmon, or extend to a large abscess formation. Free perforation occurs rarely, but fistulization does occur and most commonly to the bladder. The clinical findings vary. Most often, the clinical picture is one of fever, abdominal pain, a change in bowel habit, and localizing findings associated with leukocytosis. Computerized tomography scanning has become the procedure of choice to evaluate the symptoms since it is of less risk than a barium enema and obtains more information. The differential diagnosis may be difficult but usually can be made with accuracy. Medical treatment is preferred with appropriate antibiotic therapy and variations in fiber intake. When abscess occurs, percutaneous drainage may be tried, but when it is unsuccessful, surgical intervention is necessary. Sudden hemorrhage from a vessel in diverticula may also occur. It is estimated that approximately 20% of all patients that develop diverticula will have either inflammatory or bleeding episodes. In conclusion, fiber deficiency results in diverticular formation and a chronic inflammation that may progress to acute or chronic diverticulitis that can be treated medically but may require surgical intervention. PMID- 15115922 TI - The overlap of inflammatory bowel disease and diverticular disease. AB - An overlap of inflammatory bowel disease and diverticular disease has long been recognized. Crohn's disease and diverticulitis share clinical and radiologic features but usually differ in histopathologic findings. There is a suggestion, however, that even the characteristic pathology of Crohn's disease can be a secondary reaction to diverticulitis. It is possible, moreover, that the presence of Crohn's disease in association with diverticulosis predisposes to the development of diverticulitis. Finally, an apparently distinct form of segmental colitis associated with sigmoid diverticula has been described further contributing to the overlap of inflammatory bowel disease and diverticular disease. PMID- 15115923 TI - The pathology of diverticulosis coli. AB - Left-sided diverticulosis coli is a common condition in western communities, with 30% to 50% of adults over the age of 60 being affected. It predominantly involves the sigmoid colon. The diverticula (pseudodiverticula) are pockets of mucosa bounded by muscularis mucosae and invested with a thin layer of submucosa, that are forced out through weak points in the muscularis propria, the tips ending in the colonic subserosa. The weak points in the muscle coat are the sites of entry of the nutrient vessels of the colonic mucosa. Diverticulosis is attributed to increased colonic intraluminal pressure while straining at stool in individuals who eat low-fiber diets. Muscular hypertrophy, shortening of the bowel, and thickened mucosal folds due to mucosal redundancy are characteristic of this condition. Complications of diverticulosis include bleeding, diverticulitis, peridiverticular abscess, perforation, stricture, and fistula formation. However, most individuals with diverticulosis are asymptomatic, without evidence of complications. Mucosal changes in the diverticula in uncomplicated diverticulosis include an increased lymphoid infiltrate, development of lymphoglandular complexes, mucin depletion, mild cryptitis, architectural distortion, Paneth cell metaplasia, and ulceration. The mucosa of the remainder of the sigmoid colon (ie, the nondiverticular mucosa) is usually normal, but in about 1% of cases it has features that are indistinguishable from ulcerative colitis or from Crohn's disease (segmental colitis associated with diverticular disease, SCAD). Such cases pose a difficult diagnostic challenge as patients with SCAD respond to medical or surgical therapy for diverticular disease, whereas those with ulcerative colitis or Crohn's disease will develop other manifestations of their disease in time and require different treatment. In SCAD, the mucosal changes are confined to the area of diverticulosis; therefore, histologic evaluation of the rectum (which is unaffected by diverticulosis) and more proximal bowel can be helpful in the differential diagnosis. PMID- 15115924 TI - Diverticulitis and IBD. PMID- 15115925 TI - Microscopic colitis: pathologic considerations, changing dogma. AB - Microscopic colitis as an entity was first recognized in 1976, and has become one of the most frequent diseases to exclude on colonic mucosal biopsies. In some pathology practices, up to 30% of colonic biopsies received are from patients in whom microscopic colitis is the clinical question. In this review, the evolution of the terminology and early studies describing the pathology of microscopic colitis are discussed. The pathology of lymphocytic and collagenous colitis is reviewed in detail, including common diagnostic pitfalls, and what is currently known about the pathogenesis of these diseases. The differential diagnosis of microscopic colitis includes other idiopathic inflammatory bowel diseases (Crohn's and ulcerative colitis), infections, and drug reactions. The distinction between these entities and microscopic colitis is discussed in detail. Finally, recent studies have revealed new histopathologic changes in microscopic colitis that challenge the currently held concepts of how microscopic colitis fits into the spectrum of inflammatory bowel diseases. PMID- 15115926 TI - Diagnosis and management of microscopic colitis syndrome. AB - Microscopic colitis syndrome consists of chronic watery diarrhea, a normal or near-normal gross appearance of the colonic mucosa, and a specific histologic picture described as either lymphocytic colitis or collagenous colitis. The cause of microscopic colitis is unknown, but recent work suggests some immunologic similarities to celiac disease, suggesting that luminal antigens may be important in its pathogenesis. Diarrhea in microscopic colitis seems to be directly related to the extent of inflammation, suggesting that inflammatory mediators are responsible for reduced water absorption by the colon. Microscopic colitis is a frequent diagnosis in patients with chronic diarrhea seen at referral centers. It is often associated with other immune-mediated conditions and frequently is complicated by fecal incontinence. The differential diagnosis is broad, comprising all causes of watery diarrhea. Evaluation is straightforward with the key aspect being review of colon biopsy specimens by an experienced pathologist. Treatment is still being defined: symptomatic management with antidiarrheal agents, 5-aminosalicylate drugs, corticosteroids, especially budesonide, bile acid-binding resins, and bismuth subsalicylate all can be effective. The prognosis is good with no evidence of conversion to classic inflammatory bowel disease or of development of neoplasia over time. PMID- 15115927 TI - The clinical and pathologic significance of microscopic colitis. PMID- 15115928 TI - Postcolectomy ileitis. AB - A case is presented of a patient who had classic ulcerative colitis, both clinically and on detailed pathologic study, that underwent an ileo-pouch anal anastomosis and then developed pouchitis followed by ileitis. The patient has done fairly well on routine medications, but this case represents ileitis developing post colectomy that did not exist previous to colectomy and raises the questions for the following presentations and discussions in this workshop symposium. PMID- 15115929 TI - Classic pathology of ulcerative and Crohn's colitis. AB - The distinction between ulcerative colitis and Crohn's colitis has become crucially important since the introduction of pouch surgery. The classic pathologic findings in ulcerative colitis and Crohn's colitis are described, as well as the subtleties causing the difficulty in differentiation. In difficult cases, it is essential to review the whole patient, all available biopsy material, and the colectomy specimen. The diverted rectum in ulcerative colitis usually becomes more inflamed, whereas in Crohn's disease inflammation usually resolves. Upper gastrointestinal tract biopsies with apparent normal mucosa may also be contributory. PMID- 15115930 TI - Pathology of indeterminate colitis. AB - The term indeterminate colitis (IC) is an interim, or preliminary, descriptive term used by pathologists for cases of inflammatory bowel disease (IBD) in which a definite diagnosis of ulcerative colitis (UC) or Crohn's disease (CD) cannot be established based on the information available at the time of surgical sign-out. Most cases are due to fulminant ulcerative colitis, a condition in which the classic pathologic features of UC are often obscured and may overlap with CD. For instance, fulminant UC may show early superficial fissuring ulceration, transmural lymphoid aggregates and relative rectal sparing, simulating CD. Other common causes for establishing a diagnosis of IC include confusion of backwash ileitis in UC for terminal ileal involvement in CD, failure to accept hard criteria, such as granulomas, or segmental disease, as representative of CD, and failure to recognize unusual variants of UC that can cause CD-like patchiness of disease. Also, a diagnosis of IC should be avoided on biopsy studies and should not be established until all available clinical, endoscopic, radiologic and pathologic information are available for review. Some cases of IC may represent other forms of colitis as well, such as chronic ischemic or infectious colitis. The natural history of IC more closely resembles that of UC than CD. In fact, most cases of IC represent UC upon long-term clinical follow-up. Although, in some instances, serologic testing for ANCA or ASCA may be helpful in separating UC from CD in patients with IC, there is much overlap in the results of these assays for cases in which CD involves the colon in a UC-like pattern. Approximately 20% of IC patients develop severe pouch complications, which is intermediate in frequency between that seen in UC or CD. The risk of pouch complications, such as perianal fistulas or abscesses, and the risk of pouch breakdown is, overall, quite similar between IC and UC patients, supporting the notion that most patients with IC probably have UC and can safely undergo an ileal pouch-anal anastomosis procedure and have a reasonably good chance of having a good outcome. PMID- 15115931 TI - Indeterminate colitis: clinical spectrum of disease. AB - A diagnosis of indeterminate colitis (IC) is based on endoscopic, histologic, and radiologic findings when the criteria for either Crohn's colitis or ulcerative colitis (UC) cannot be definitively established. Population-based studies have demonstrated that the average annual incidence of IC ranges 1.6 to 2.4/100,000 versus 7.3 to 13.6/100,000 for UC. At the time of initial diagnosis of inflammatory bowel disease, up to 10% to 15% of patients will be diagnosed as having IC. Over time, greater than 50% of patients with IC will be given a diagnosis of UC or Crohn's, with the majority being diagnosed with UC. Therefore, approximately 4% to 5% of all patients with inflammatory bowel disease will be left with the diagnosis of IC. Clinically, patients with IC seem to have a more severe course with a greater chance of colectomy and pouch failure. No studies have been undertaken to determine the optimum treatment regimen for IC. Recent studies have shown that patients with IC are still appropriate candidates for ileal pouch anal anastomosis. In conclusion, the current data support the premise that IC may be a separate entity, but future studies will have to focus on the genotypic and phenotypic characterization of these patients. PMID- 15115932 TI - The diagnosis and treatment of pouchitis in inflammatory bowel disease. AB - The ileal pouch anal anastomosis (IPAA) procedure has become the preferred surgical option for most patients with ulcerative colitis who require surgical removal of the colorectum. The vast majority of patients with this new anatomy will either not develop pouchitis or develop a few discrete episodes of acute pouchitis. However approximately one fourth of patients will develop recurrent pouchitis, with 5% being categorized as chronic pouchitis requiring maintenance therapy or, on rare occasion, pouch excision. Factors that are associated with an increased risk of pouchitis include primary sclerosing cholangitis, extraintestinal manifestations, and nonsmokers. Controversy surrounds other risk factors such as extent of colitis, backwash ileitis, preoperative pANCA levels, and carrying a specific allele for IL-1 receptor antagonist. The etiology of pouchitis is unknown, but theories range from genetic susceptibility, bacterial overgrowth, ischemia, and fecal stasis, to a recurrence of ulcerative colitis in the pouch, a missed diagnosis of Crohn's disease, or possibly a novel third form of inflammatory bowel disease. Some patients with symptoms of pouchitis will not have inflammation of the pouch, but rather, irritable pouch syndrome. Thus, endoscopic investigation with biopsy is important for declaring whether a patient has pouchitis. Indeed, the more commonly used scores, such as the pouch disease activity index, incorporate both endoscopic and histologic criteria. Not surprisingly, treatment options for patients with pouchitis resemble that of regular inflammatory bowel disease, although there have only been a few controlled trials. Antibiotics are the mainstay of therapy, with metronidazole and ciprofloxacin demonstrating benefit in controlled trials. Probiotics are effective for maintaining remission of pouchitis. Mesalamine, corticosteroids, and immunomodulators have been used with some success. Occasionally, patients with well-documented ulcerative colitis as the indication for IPAA will develop what appears to be Crohn's disease of the pouch, on the basis of granulomatous inflammation, pre-pouch ileitis, or fistulae. The treatment is similar to Crohn's disease, including the use of infliximab. Dysplasia within the pouch mucosa itself is quite rare. Reports of dysplasia occurring in patients with IPAA are usually due to neoplastic change within the residual cuff of rectal or transition zone mucosa just below the pouch, rather than in the ileal mucosa of the pouch. With further elucidation of the genetic basis for inflammatory bowel disease, we should be able to more accurately classify patients with ulcerative colitis and Crohn's disease genotypically. Hopefully, this will also bring more clarity to the heterogeneous population of patients with pouchitis and allow for more focused therapeutic strategies. PMID- 15115933 TI - Do serological markers and cytokines determine the indeterminate? AB - There is a general sense that indeterminate colitis (IC) runs an aggressive clinical course, is medically refractory, and is associated with higher pouch failure rates following restorative proctocolectomy. The question has been raised whether IC can be assigned to a diagnosis of Crohn's disease or ulcerative colitis through the characterization of immunogenetic similarities, or whether IC may represent a distinct subgroup within these heterogeneous disorders. In this article, the use of serologic markers, genetics, and immune responses to understand the pathogenesis of inflammatory bowel disease (IBD) and define clinically important subgroups of patients will be discussed. Then, how these scientific advances have been applied to the entity of IC will be reviewed. Importantly, a recent prospective study suggests that the absence of IBD associated serologic markers defines the majority of IC as a separate entity within the spectrum of IBD. The development of serologic, genetic, and immune response markers will allow for rational description of clinically important subgroups, redefine natural history, and predict responses to therapy in IBD. PMID- 15115934 TI - Indeterminate colitis. PMID- 15115943 TI - Taking another look at schizophrenia. PMID- 15115945 TI - Social cognition and its neural correlates in schizophrenia and autism. AB - The study of social cognition in psychiatric disorders has become increasingly popular in recent years. This is due to the its proposed link to social functioning and the inability of general neurocognitive skills to explain the spectrum of impairments observed in patients. This article reviews research into two of the processes thought to underlie social cognition (emotion perception and theory of mind) in schizophrenia and autism. This is followed by a look at neuroimaging studies and their efforts to localize the neural correlates of emotion perception and theory of mind in the two disorders. We concluded that while a specific impairment in emotion perception and theory of mind skills cannot be generalized to all individuals with autism and schizophrenia, there are subpopulations that have lingering deficits of social cognition tasks. Neuroimaging work consistently points to the involvement of the fusiform gyrus and amygdala in emotion processing, while the medial prefrontal and frontal cortex are implicated in tasks invoking theory of mind. We propose that deficits of social cognition may benefit from cognitive remediation therapy and pharmacological cognitive enhancers. PMID- 15115946 TI - Trail making and olfaction in schizophrenia: implications for processing speed. AB - BACKGROUND: Previous research has established a relationship between smell identification deficits (SID) and particular aspects of cognitive function among patients with schizophrenia. OBJECTIVE: To expand the extant literature, we examined the relationship between SID and the Trail Making Test to determine if processing speed is related to SID. METHODS: Our sample included 60 inpatients from the New York State Psychiatric Institute's Schizophrenia Research Unit. We considered age, deficit syndrome, verbal intelligence quotient, and education in our analyses due to their documented relationship to smell identification ability. RESULTS: Trails A errors and Trails A seconds accounted for a significant amount of the variance in University of Pennsylvania Smell Identification Test scores in a regression analysis (R2=.10, P=.008 and R2=.05, P=.04). CONCLUSION: Linking neurocognition to smell identification deficits may prove to be an essential marker for schizophrenia research. PMID- 15115947 TI - Potential noradrenergic targets for cognitive enhancement in schizophrenia. AB - Substantial evidence suggests that alterations in noradrenergic function contribute to the cognitive impairments of schizophrenia. Activation of post junctional alpha 2a-adrenergic receptors in the prefrontal cortex by the alpha 2a selective agonist guanfacine has demonstrated some preliminary benefit in subjects with schizophrenia treated with atypical antipsychotics. alpha 1 adrenergic receptor activity may be less important in mediating the cognitive impairments of schizophrenia. beta-adrenergic receptors may serve as another potential target for cognitive remediation in schizophrenia. However, the potential increase in memory consolidation in schizophrenia patients produced by beta-adrenergic agonists may be outweighed by the impairment in cognitive flexibility and executive functioning produced by beta-adrenergic agonists. Finally, norepinephrine reuptake inhibitors, such as atomoxetine, hold promise as potential cognitive enhancers in schizophrenia because of their ability to indirectly but selectively increase extracellular dopamine concentrations in the prefrontal cortex. PMID- 15115948 TI - Improvement in prosocial functioning after a switch to ziprasidone treatment. AB - BACKGROUND: Cognitive, social, and affective impairments are major features of schizophrenia, despite not being represented in the formal diagnostic criteria. These impairments are associated with major reductions in quality of life for patients with schizophrenia. Treatment with newer antipsychotic medications has been reported to improve all of these areas of functioning, but most studies have not examined the direct association between changes in cognitive functioning and other aspects of the illness. OBJECTIVES: The goal of this analysis was to examine relationships between cognitive and affective symptoms and their impact on social impairments in patients switched to ziprasidone treatment. METHODS: In this study, which is a re-analysis of previously published data, 270 patients were switched from previous treatment with conventional antipsychotics, risperidone, or olanzapine to treatment with ziprasidone. Patients were tested with a cognitive assessment battery, rated with the Positive and Negative Syndrome Scale (PANSS), and received other assessments of safety and tolerability. PANSS scores were divided into factors based on previously published factor analyses, with a focus on the cognitive, prosocial, and anxiety depression factors. RESULTS: Statistically significant improvements for global cognitive functioning and the three PANSS subscales were found across the studies. When the data were pooled for a path analysis, changes in cognitive functioning indexed by the PANSS cognitive subscales was the primary predictor of improvements in PANSS prosocial functions, with changes in anxiety-depression accounting for much less variance. CONCLUSION: These results suggest a direct relationship between improvements in cognitive and prosocial functioning in patients with schizophrenia and indicate that treatments that enhance cognitive functioning, such novel antipsychotics, have the potential to improve aspects of outcome in schizophrenia even in short-term treatment studies. This issue should be addressed in future double-blind studies of the effects of atypical medications in schizophrenia. PMID- 15115949 TI - Effects of low-dose risperidone and low-dose zuclopenthixol on cognitive functions in first-episode drug-naive schizophrenic patients. AB - BACKGROUND: Studies on the effects of antipsychotics on cognitive deficits in schizophrenia mostly suggest a superior effect of atypical over typical compounds, although findings are inconsistent and effect sizes small. Several methodological issues, such as heterogeneous patient samples, incomparable drug doses, effects of prior medication, construct validity, and retest effects on neuropsychological tasks, confound most results and the comparability between studies. Consequently, the conclusion concerning effects of antipsychotics on cognition is still equivocal. OBJECTIVE: The present randomized clinical trial examined the effects on cognition of comparatively low doses of a typical antipsychotic (zuclopenthixol) and an atypical antipsychotic (risperidone) in a homogeneous group of drug-naive first-episode schizophrenic patients in a longitudinal setting. METHODS: First-episode schizophrenic patients who had never previously been exposed to antipsychotic treatment (N=25) were randomly allocated to treatment with flexible doses of zuclopenthixol or risperidone in an open label design. Cognitive functions were examined both when patients were drug naive, and after 13 weeks of treatment. A comprehensive neuropsychological battery was used in order to optimize construct validity, and principal components of cognitive functions were extrapolated in order to reduce type I errors. A healthy control group was tested at baseline and after 13 weeks, in order to examine retest effects. The cognitive domains studied were executive functions, selective attention, and reaction time. RESULTS: The patients showed considerable cognitive deficits when drug-naive. There were few differential effects of risperidone and zuclopenthixol on cognitive deficits, except for a differential significance, respectively, tendency towards improved reaction and movement times in the risperidone group, and a lack of such in the zuclopenthixol group. These differences were no longer significant after covarying for extrapyramidal side effects and anticholinergic medication that were more prevalent in the zuclopenthixol group and the increases after medication were comparable with retest effects in controls. CONCLUSION: The study underscores the importance of examining impact of factors, such as clinical improvement, extrapyramidal side effects, anticholinergic medication and retest effects in longitudinal efficacy studies. This study does not support efficacy of either risperidone or zuclopenthixol on cognitive functions in drug-naive schizophrenia patients after 3 months of medication, because neither could be distinguished from retest effects of the healthy control group. PMID- 15115950 TI - Repetitive transcranial magnetic stimulation improves depersonalization: a case report. AB - Depersonalization disorder is a poorly understood and treatment-resistant condition. This report describes a patient with depersonalization disorder who underwent six sessions of repetitive transcranial magnetic stimulation on the left dorsolateral prefrontal cortex. Repetitive transcranial magnetic stimulation produced a 28% reduction on depersonalization scores. PMID- 15115951 TI - The clinical efficacy and safety of galantamine in the treatment of Alzheimer's disease. AB - Alzheimer's disease is a progressive, neurodegenerative condition characterized by deficits in cognition, inability to perform activities of daily living, and alterations in behavior. Galantamine hydrobromide is the newest acetylcholinesterase inhibitor (AChEI) approved in the United States for the treatment of mild-to-moderate AD. The safety and efficacy of galantamine has been demonstrated in multiple randomized, Phase III trials of >2,600 patients with mild-to-moderate AD. Studies have found that galantamine improved or maintained performance in all domains of AD (cognition, function, behavior, and caregiver burden) in the short term and slowed the decline in performance or maintained baseline performance through 12 months. The dual mechanism of action may make galantamine a reasonable treatment option for both newly diagnosed patients and patients who have not benefitted from or have poorly tolerated current therapy. PMID- 15115953 TI - Cholestasis: human disease and experimental animal models. AB - Cholestasis may result from a failure in bile secretion in hepatocytes or ductular cells, or from a blockade to the free bile flow. Human cholestasis may be induced by many drugs, being antibiotics the more common. Other types of cholestasis seen in humans are a group of familial cholestatic disorders, obstructive cholestasis, primary biliary cirrhosis, extrahepatic biliary atresia, primary sclerosing cholangitis, cholestasis of pregnancy, oral contraceptive induced cholestasis, and sepsis-induced cholestasis. Experimental animal models allow the understanding of pathophysiological mechanisms involved and their clinical correlates. The most common experimental models of intrahepatic cholestasis are estrogen-induced, endotoxin-induced and drug-induced cholestasis. A well known model of extrahepatic biliary obstruction is common bile duct ligation. Drug-induced cholestasis were described using different drugs. On this regard, alpha naphthylisothiocyanate treatment has been extensively used, permitting to describe not only cholestatic alterations but also compensatory mechanisms. Congenital defficiency of transport proteins also were studied in natural rat models of cholestasis. The experimental animal models allow to define down-regulated alterations of hepatocyte transport proteins, and up-regulated ones acting as compensatory mechanisms. In conclusion, animal model and transport protein studies are necessary for the progressive understanding of congenital and acquired human cholestasis, and regulatory mechanisms that operate on liver cells. PMID- 15115954 TI - Liver fibrosis and inflammation. A review. AB - Hepatic fibrosis, is a wound healing process characterized by accumulation of extracellular matrix proteins (ECM) especially collagen types I and III, as well as an increase in other extracellular matrix constituents such as proteoglycans, fibronectin and laminin in response to liver injury. Recruitment of leukocytes takes place after the insult and requires several adhesion molecules. Monocytes and macrophages are involved in inflammatory actions by producing nitric oxide and inflammatory cytokines. As a consequence of chronic tissue damage stellate cells (SC) as well as extracellular matrix producting cells, undergo a process of activation characterized by proliferation, motility, contractility, and synthesis of extracellular matrix. Activation of SC is regulated by several soluble factors, including cytokines, chemokines, growth factors, and products of oxidative stress. TGF - b and IL- 6 are the two main fibrogenic cytokines. Potential regulatory factors of the activation of SC are important targets for future antifibrogenic treatments. PMID- 15115955 TI - New therapies on the horizon for hepatitis C. AB - Therapy of chronic HCV infection has greatly improved in recent years with the addition of ribavirin to alpha interferon and has further improved more with the use of PEG-interferons. However, more than half of patients do not achieve lasting benefits from these therapies. The future therapeutic developments may include one or more of the following approaches: understanding the HCV genomic organization, elucidating the viral life cycle and HCV replication strategy and understanding the immune mechanisms required for viral propagation or infectivity. The development of novel antiviral strategies and a preventive vaccine against HCV infection remains a major challenge for the future, and will depend on progress on both molecular biology as well as clinical studies. Unfortunately, the low replication of the virus in culture, the lack of convenient animal models, and the high genome variability present mayor challenges for drug development. PMID- 15115956 TI - Transthyretin familial amyloidotic polyneuropathy: histopathological study of the explanted livers. AB - FAP is an autosomal dominant inherited disease, characterized by systemic deposition of amyloid fibrils in various tissues. The purpose of this study is to describe the gross and microscopic findings of the explanted livers for FAP.10 patients were transplanted for FAP at our institution. Diagnosis was supported by positive familiar history, clinical data and detection of mutated TTR by electrospray ionization mass spectrometry with Val30Met mutation verified by PCR. All the explanted livers were photographed, fixed in formol and processed according to protocol. Later they were examined with HyE, reticulin, PAS diastasa, Masson trichromic, Congo red with polarised light and immunoreactivity against TTR. The gross aspect was normal. We obtained multiple samples representative of the organ and the hepatic hilium. All of the patients presented with deposits of amyloid substance in the lymph nodes and the nerves of the hepatic hilium These deposits were Congo red positive with a greenish birefringence to polarized light Deposits show immunoreactivity with antihuman TTR. Whereas liver transplantation restores hepatic function in patients with cirrhosis, liver transplantation cures the FAP patient of their genetic defect. Domino transplantation is a procedure in which the index patient receives an organ, while the explanted organ is reused for transplantation into another patient. In conclusion, exclusion of hepatic amyloid deposits which can cause functional alterations in the FAP liver is vital; and is important to study the explanted livers of patients with FAP to confirm the results of the scarce published series. PMID- 15115957 TI - Ketorolac pharmacokinetics in experimental cirrhosis by bile duct ligation in the rat. AB - The purpose of the present work was to study the pharmacokinetics of ketorolac, a poorly metabolized drug, in experimental cirrhosis. Cirrhosis was induced by bile duct ligation (BDL) for four weeks in male Wistar rats. Ketorolac was given intravenously (1 mg/kg ) or orally (3.2 mg/kg) to control (sham-operated) and BDL rats. Determination of ketorolac in plasma was carried out by HPLC and estimation of pharmacokinetic parameters was performed by non-compartmental analysis. Indicators of liver damage and liver fibrosis were significantly increased (p < 0.05) in BDL compared to control rats. Experimental cirrhosis did not induce any significant alteration in intravenous ketorolac pharmacokinetics. Volume of distribution, clearance, AUC and t1/2 were similar in BDL and control animals. Notwithstanding, oral ketorolac bioavailability was significantly altered in BDL rats. AUC and Cmax were reduced, while tmax was prolonged, suggesting that both, the extent and the rate of ketorolac absorption were decreased. Results show that liver cirrhosis may result in significant pharmacokinetic alterations, even for poorly bio-transformed drugs, but that alterations may vary with the route of administration. In conclusion, uncritical generalizations on the effect of liver damage on drug kinetics should be avoided and systematic studies for every drug and every route of administration are thus recommended. PMID- 15115959 TI - Hypertransaminasemia and severe hepatic steatosis without inflammation. A case report. AB - Nonalcoholic fatty liver disease (NAFLD) is a medical condition that may progress to end-stage liver disease. The spectrum of NAFLD is wide and ranges from simple fat accumulation in hepatocytes (steatosis), to fat accumulation plus necroinflammatory activity with or without fibrosis (steatohepatitis). In addition, NAFLD is the most common cause of abnormal liver-test results among adults with a prevalence of 13%-23%. This case report is an example of a patient with asymptomatic hypertransaminasemia and severe hepatic steatosis without inflammation in which the diagnosis was made by liver biopsy. PMID- 15115958 TI - Hepatic fascioliasis. PMID- 15115961 TI - Current therapies for chronic hepatitis B. PMID- 15115962 TI - New concepts of mechanisms of intestinal cholesterol absorption. AB - The small intestine is a unique organ providing dietary and reabsorbed biliary cholesterol to the body. However, the molecular mechanisms whereby cholesterol is absorbed have not yet been fully understood. Recent research suggests that the newly identified ATP-binding cassette (ABC) transporters ABCG5 and ABCG8 are apical cholesterol export pumps that promote partial efflux of cholesterol and nearly complete efflux of plant sterols from enterocytes into the intestinal lumen after their absorption. This provides an explanation why cholesterol absorption is a selective process in that plant sterols and other non-cholesterol sterols are absorbed poorly or not at all. Furthermore, a putative cholesterol import protein has been proposed, but remains uncharacterized. The identification of such a gene should yield new insights into the mechanisms that potentially regulate the influx of cholesterol across the apical brush border membrane of the enterocyte. Combination therapy using a novel and potent cholesterol absorption inhibitor (ezetimibe) and an HMG-CoA reductase inhibitor (statins) offers an efficacious new approach to the prevention and treatment of hypercholesterolemia. PMID- 15115964 TI - Liver transplantation for Wilson s disease: our experience with review of the literature. AB - BACKGROUND/AIMS: Orthotopic liver transplantation is being used with more frequency as the treatment for Wilson s disease. The experience at the Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran with orthotopic liver transplantation for Wilson s disease is reported. We perform an extensive literature review for this treatment modality. METHODS: Between january 2000 and june 2003, 23 orthotopic liver transplants were performed at our institution, 2 of them for Wilson s disease. Both the patients presented with chronic advanced liver disease and one presented neurologic dysfunction. RESULTS: Both the patients were transplanted without any major complication and are alive 43 and 22 months after the transplant respectively. To our knowledge 370 liver transplants have been reported in the international literature since 1994 for the treatment of Wilson s disease. CONCLUSIONS: Currently, orthotopic liver transplantation should be considered as a major option for the treatment of chronic liver disease in patients with Wilson s disease. Although it is well known that the transplant only partially corrects the defective metabolism in patients with Wilson s disease, it does convert the copper kinetics of a homozygous to that of a heterozigote, thus, providing an effective phenotypic cure. PMID- 15115963 TI - Hepatic encephalopathy: a review. AB - Hepatic encephalopathy (HE) is a complication that presents in as many as 28% of patients with cirrhosis, and reported up to ten years after the diagnosis of cirrhosis. Commonly, it is observed in patients with severe hepatic failure and is characterized by neuropsychiatric manifestations that can range in severity from a mild alteration in mental state to a coma; additionally, some neuromuscular symptoms can be observed. This complication of either acute or chronic hepatic disease is the result of a diminished hepatic reservoir and inability to detoxify some toxins that originate in the bowel. Today, the role of astrocytes, specifically the Alzheimer type II cells, is known to be very important in the pathogenesis of the hepatic encephalopathy, and will be reviewed later. In conclusion, the objectives of this review are: To understand the pathogenesis of hepatic encephalopathy, To recognize the precipitating factors, as well as preventive measures for the development of the hepatic encephalopathy, To describe the new classification of hepatic encephalopathy and its clinical implications, To recognize the clinical manifestations and stages of the disease, To understand the main diagnostic tests used to detect the hepatic encephalopathy, To describe the main therapeutic treatments of hepatic encephalopathy. PMID- 15115965 TI - Peginterferon alfa-2a plus ribavirin for treating chronic hepatitis C virus infection: analysis of Mexican patients included in a multicenter international clinical trial. AB - Treatment with polyethylene glycol-modified interferon alfa-2a (peginterferon) alone produces significantly higher sustained antiviral responses than treatment with interferon alfa-2a alone in patients with chronic hepatitis C virus (HCV) infection. We compared the efficacy and safety of peginterferon alfa-2a plus ribavirin, interferon alfa-2b plus ribavirin, and peginterferon alfa-2a alone in the initial treatment of chronic hepatitis C. Thirty-two patients were randomly assigned to treatment, and received at least one dose of medication consisting of 180 microg of peginterferon alfa-2a once weekly plus daily ribavirin (1,000 or 1,200 mg, depending on body weight) (n = 14), weekly peginterferon alfa-2a plus daily placebo (n = 6), or three million units of interferon alfa-2b thrice weekly plus daily ribavirin for 48 weeks (n = 12). More patients who received peginterferon alfa-2a plus ribavirin had a sustained virologic response (defined as the absence of detectable HCV RNA 24 weeks after cessation of therapy) than patients who received interferon alfa-2b plus ribavirin (7/14 vs. 4/12) or peginterferon alfa-2a plus placebo (0/6). The overall safety profiles of the three treatment regimens were similar. In conclusion, for patients with chronic hepatitis C, once-weekly peginterferon alfa-2a plus ribavirin was tolerated as well as interferon alfa-2b plus ribavirin and produced significant improvements in the rate of sustained viral reduction compared with interferon alfa-2b plus ribavirin or peginterferon alfa-2a alone. PMID- 15115966 TI - "Tear-drop shaped" HCC. PMID- 15115967 TI - Hemobilia. A case report. AB - We present the case of a 88 years old male, with a history of melena, demonstrated by repeated endoscopies to be secondary to hemobilia, and after endoscopic retrograde cholangiopancreatography we could demonstrate the presence of bleeding intraductal choledocus polyps with histopathological report of intraepithelial adenoma. Hemobilia is a rare cause of upper gastrointestinal hemorrhage with an increasing incidence because of the widespread use of invasive hepatobiliary procedures and improved recognition. In the majority of cases the cause is iatrogenic. The classical presentation of hemobilia is with biliary colic, jaundice, hematemesis, and melena. The diagnosis and evaluation of hemobilia is facilitated by the use of sonography, computed tomography and endoscopic retrograde cholangiopancreatography. Persistent bleeding sometimes requires urgent therapeutic intervention, such as angiography or surgery. PMID- 15115969 TI - Dynamics of hepatitis C virus infection. AB - Viremia shows only minor fluctuations in untreated patients chronically infected with hepatitis C virus. The steady state situation of balanced viral production and clearance in untreated patients can be disturbed by active antiviral treatment. After initiating interferon- alpha therapy, a typical biphasic decline of viremia can be observed and analyzed. Evaluation of mathematical models of viral dynamics during the initial phase of antiviral treatment shows high turnover rates of pre-treatment viral production and clearance of about 10(11) 10(13) virions each day and in-vivo half-lives of a few hours for free hepatitis C virions. During the first 24 to 48 hours of therapy, a dose-dependent first phase of interferon-alpha induced viral kinetics is characterized by a rapid exponential decline of serum viral load. Then viral decline enters a second phase of a relatively slow exponential decay during the following weeks of therapy which mainly reflects the death rate of infected hepatocytes. This second phase decay is predictive for the virologic end-of-treatment and even more the sustained response. Non-responding patients typically show constant viremia or even a rebound during this second phase. PMID- 15115970 TI - From blood to bile: recent advances in hepatobiliary transport. AB - Transport of endogenous and exogenous substances from blood to bile is an essential function of the liver. In the last decade a still growing number of specific transport proteins present at the sinusoidal and canalicular membrane domains of hepatocytes and cholangiocytes have been cloned and functionally characterized. Studies assessing the molecular expression and function of these hepatobiliary transport proteins under different experimental conditions has helped to define the adaptive responses of hepatocytes to certain physiological states and to cholestatic liver injury and to a better understanding of the physiology of bile formation and of the pathophysiology of certain cholestatic diseases. Particularly relevant is the elucidation of the molecular bases of several forms of inherited cholestatic liver disease, which may help to the development of better diagnostic tools or to the design of new therapeutic strategies. In the present review we summarize recent experimental and clinical data involving hepatobiliary transport mechanisms. PMID- 15115971 TI - Pathophysiology, diagnosis and treatment of ascites in cirrhosis. AB - The mechanism by which ascites develops in cirrhosis is multifactorial Severe sinusoidal portal hypertension and hepatic insufficiency are the initial factors. They lead to a circulatory dysfunction characterized by arterial vasodilation, arterial hypotension, high cardiac output and hypervolemia and to renal sodium and water retention. There are evidences that arterial vasodilation in cirrhosis occurs in the splanchnic circulation and is related to an increased synthesis of local vasodilators. Vascular resistance is normal or increased in the remaining major vascular territories (kidney, muscle and skin and brain). Splanchnic arterial vasodilation not only impairs systemic hemodynamics and renal function but also alters hemodynamics in the splanchnic microcirculation. The rapid and high inflow of arterial blood into the splanchnic microcirculation is the main factor increasing hydrostatic pressure in the splanchnic capillaries leading to an excessive production of splanchnic lymph over lymphatic return. Lymph leakage from the liver and other splanchnic organs is the mechanism of fluid accumulation in the abdominal cavity. Continuous renal sodium and water retention perpetuates ascites formation. Large volume paracentesis associated with albumin infusion is the treatment of choice of tense ascites because it is very effective and rapid and is associated with fewer complications that the traditional treatment (sodium restriction and diuretics). However, diuretic should be given after paracentesis to prevent reaccumulation of ascites. In patients with moderate ascites diuretics should be preferred as initial therapy. Patients with refractory ascites could be treated by paracentesis or percutaneous transjugular portacaval shunt (TIPS). TIPS is more effective in the long term control of ascites but may impair hepatic function and induce chronic hepatic encephalopathy. PMID- 15115972 TI - Development of a liver unit in Latin America. AB - The Liver Unit at the "Dr. Jose E. Gonzalez" University Hospital and School of Medicine of the Autonomous University of Nuevo Leon in Monterrey was founded in 1983. Over the years, it has become a referral center for the northeast of Mexico. The frequency of diagnosis has changed: in 1983, the most common liver disease seen was alcoholic liver disease, today it is chronic hepatitis C. Amebic liver abscess, which used to be common, was hardly seen in 2001. Non-alcoholic fatty liver disease was unidentified 18 years ago, whereas in 2001 it was seen in 10% of patients. The development of five laboratories within the unit has allowed us to implement basic and clinical research trials, and to offer a high quality diagnostic service. The experimental liver transplant program started in 1987 and a clinical program in humans in 1991: four patients received an orthotopic liver transplantation in its first phase. In the second phase, 20 patients received allografts from September 1999 to March 2002. Technical complications have been encountered in only one patient, with a biliary leak, and there have been three perioperative deaths. Infections occurred in eight patients; all resolved. Acute postoperative rejection occurred in two patients, and in the first seven months in another five; all of them resolved. The two-year survival rate is 80%. This unit offers a highly specialized diagnosis, standardized specialized laboratory services and a transplant program that guarantees a higher quality of medical attention to patients with liver diseases. PMID- 15115973 TI - Hepatic apolipoprotein A-I gene expression in patients with cholesterol gallstones treated with ursodeoxycholic acid. AB - OBJECTIVE: It has been suggested that apo A-I can inhibit cholesterol crystal nucleation in vitro, and ursodeoxycholic acid (UDCA) is a safe and effective treatment for selected patients with cholesterol gallstones the aim of this study was to investigate the effect of UDCA on the steady-state levels (SSL) of apo A-I mRNA in the liver, as well as serum apo A-I, in patients with cholesterol gallstones. DESIGN: Twenty Mexican patients with symptomatic radiolucent gallstones were randomized and assigned in a double blind fashion to groups that were administered either UDCA (4 mg/kg per day) or placebo for 10 to 15 days before cholecystectomy. Apo A- I mRNA levels in liver and gallbladder tissues were determined by northern blot and serum levels of apo A- I by turbidimetric method. METHODS: Apo A- I mRNA levels were higher in nine of the 10 patients who received UDCA and in comparison to those to the placebo group. In the gallbladder apo A- I mRNA levels were undetected. Serum levels (mg/dL) of apo A- I were similar in both UDCA and placebo groups after treatment (111.7 +/- 29.8 vs 115.6 +/- 25.4). CONCLUSIONS: The results of this study shown that apo A- I mRNA gene express at the mRNA level in the liver but not in the gallbladder of patients with cholesterol gallstones treated with UDCA. PMID- 15115974 TI - Amebic liver abscess open to the inferior vena cava. PMID- 15115975 TI - Epithelioid granulomas in a patient with hepatitis C virus. AB - Hepatitis C virus infection causes an epidemic disease. The morphologic aspects of hepatitis C infection (HCV) are well established with regards to necroinflammatory processes and consequences like fibrosis, cirrhosis, and related neoplasms. However, the presence of epithelioid granulomas has not been well described for this infection. We report a patient with HCV and granulomas without any other co-infection or history of drug abuse. PMID- 15115976 TI - Who moved my heart? Adaptive responses to disruptive challenges. PMID- 15115977 TI - Heart failure overview. PMID- 15115978 TI - Is it important to examine gender differences in the epidemiology and outcome of severe heart failure? PMID- 15115979 TI - Transplant and ventricular assist devices: gender differences in application and implementation. PMID- 15115980 TI - The Boston Circulatory Arrest Study: an analysis. PMID- 15115981 TI - Ethyl pyruvate preserves cardiac function and attenuates oxidative injury after prolonged myocardial ischemia. AB - OBJECTIVE: Myocardial injury and dysfunction following ischemia are mediated in part by reactive oxygen species. Pyruvate, a key glycolytic intermediary, is an effective free radical scavenger but unfortunately is limited by aqueous instability. The ester derivative, ethyl pyruvate, is stable in solution and should function as an antioxidant and energy precursor. This study sought to evaluate ethyl pyruvate as a myocardial protective agent in a rat model of ischemia-reperfusion injury. METHODS: Rats underwent 30-minute ischemia and 30 minute reperfusion of the left anterior descending coronary artery territory. Immediately prior to both ischemia and reperfusion, animals received an intravenous bolus of either ethyl pyruvate (n = 26) or vehicle control (n = 26). Myocardial high-energy phosphate levels were determined by adenosine triphosphate assay, oxidative injury was measured by lipid peroxidation assay, infarct size was quantified by triphenyltetrazolium chloride staining, and cardiac function was assessed in vivo. RESULTS: Ethyl pyruvate administration significantly increased myocardial adenosine triphosphate levels compared with control (87.6 +/ 29.2 nmol/g vs 10.0 +/- 2.4 nmol/g, P =.03). In ischemic myocardium, ethyl pyruvate reduced oxidative injury compared with control (63.8 +/- 3.3 nmol/g vs 89.5 +/- 3.0 nmol/g, P <.001). Ethyl pyruvate diminished infarct size as a percentage of area at risk (25.3% +/- 1.5% vs 33.6% +/- 2.1%, P =.005). Ethyl pyruvate improved myocardial function compared with control (maximum pressure: 86.6 +/- 2.9 mm Hg vs 73.5 +/- 2.5 mm Hg, P <.001; maximum rate of pressure rise: 3518 +/- 243 mm Hg/s vs 2703 +/- 175 mm Hg/s, P =.005; maximal rate of ventricular systolic volume ejection: 3097 +/- 479 microL/s vs 2120 +/- 287 microL/s, P =.04; ejection fraction: 41.9% +/- 3.8% vs 31.4% +/- 4.1%, P =.03; cardiac output: 26.7 +/- 0.9 mL/min vs 22.7 +/- 1.3 mL/min, P =.01; and end systolic pressure-volume relationship slope: 1.09 +/- 0.22 vs 0.59 +/- 0.2, P =.02). CONCLUSIONS: In this study of myocardial ischemia-reperfusion injury, ethyl pyruvate enhanced myocardial adenosine triphosphate levels, attenuated myocardial oxidative injury, decreased infarct size, and preserved cardiac function. PMID- 15115982 TI - Safety of deliberate intraoperative and postoperative hypothermia for patients undergoing coronary artery surgery: a randomized trial. AB - BACKGROUND: Hypothermia in the perioperative period is associated with adverse effects, particularly bleeding. Before termination of cardiopulmonary bypass, rewarming times and perfusion temperatures are often increased to avoid post cardiopulmonary bypass hypothermia and the presumed complications. This practice may, however, also have adverse effects, particularly cerebral hyperthermia. We present safety outcomes from a trial in which patients undergoing coronary artery surgery were randomly assigned to normothermia or hypothermia for the entire surgical procedure. METHODS: Consenting patients over the age of 60 years presenting for a first, elective coronary artery surgery with cardiopulmonary bypass were randomly assigned to having their nasopharyngeal temperature maintained at either 37 degrees C (group N; 73 patients) or 34 degrees C (group H; 71 patients) throughout the intraoperative period, with no rewarming before arrival in the intensive care unit. All received tranexamic acid. RESULTS: There was no clinically important difference in intraoperative blood product or inotrope use. Temperatures on arrival in the intensive care unit were 36.7 degrees C +/- 0.38 degrees C and 34.3 degrees C +/- 0.38 degrees C in groups N and H, respectively. Blood loss during the first 12 postoperative hours was 596 +/- 356 mL in group N and 666 +/- 405 mL in group H (mean difference +/- 95% confidence interval, 70 +/- 126 mL; P =.28). There was no significant difference in blood product utilization, intubation time, time in the hospital, myocardial infarction, or mortality. The mean time in the intensive care unit was 8.4 hours less in the hypothermic group (P =.02). CONCLUSIONS: Our data support the safety of perioperative mild hypothermia in patients undergoing elective nonreoperative coronary artery surgery with cardiopulmonary bypass. These findings suggest that complete rewarming after hypothermic cardiopulmonary bypass is not necessary in all cases. PMID- 15115983 TI - Vein graft arterialization causes differential activation of mitogen-activated protein kinases. AB - OBJECTIVE: Vascular injury results in activation of the mitogen-activated protein kinases-extracellular-signal regulated kinases, c-jun N-terminal kinase, and p38(MAPK)-which have been implicated in cell proliferation, migration, and apoptosis. The goal of this study was to characterize mitogen-activated protein kinase activation in arterialized vein grafts. METHODS: Carotid artery bypass using reversed external jugular vein was performed in 29 dogs. Vein grafts were harvested after 30 minutes and 3, 8, and 24 hours, and 4, 7, 14, and 28 days. Contralateral external jugular vein and external jugular vein interposition vein to-vein grafts were used as controls. Vein graft extracts were analyzed for extracellular-signal regulated kinases, c-jun N-terminal kinase, and p38(MAPK) activation. Proliferating cell nuclear antigen expression was investigated as a parameter of cell proliferation. Apoptosis was assessed by terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate nick end labeling staining and intimal hyperplasia by morphometric examination of tissue sections. RESULTS: Significant intimal hyperplasia was observed at 28 days. Over the time points studied, vein graft arterialization resulted in bimodal activation of both extracellular-signal regulated kinase and p38(MAPK) (30 minutes through 3 hours; 4 days) but did not induce activation of c-jun N terminal kinase. Proliferating cell nuclear antigen expression increased from days 1 through 28, and apoptosis increased between 8 and 24 hours. CONCLUSION: Vein graft arterialization induces bimodal activation of extracellular-signal regulated kinase and p38(MAPK); however, in contrast with what is described in arterial injury, it does not induce c-jun N-terminal kinase activation. These results provide the first comprehensive characterization of the mitogen-activated protein kinase signaling pathways activated in vein graft arterialization and identify mitogen-activated protein kinases as potential mediators of vein graft remodeling and subsequent intimal hyperplasia. PMID- 15115984 TI - Inhaled nitric oxide decreases pulmonary soluble guanylate cyclase protein levels in 1-month-old lambs. AB - BACKGROUND: Inhaled nitric oxide produces potent pulmonary vasodilation by activating soluble guanylate cyclase and increasing smooth muscle cell concentrations of cyclic guanosine monophosphate. However, responses are often nonsustained, and clinically significant increases in pulmonary vascular resistance have been noted on its acute withdrawal. In vitro and in vivo data suggest that inhaled nitric oxide decreases endogenous nitric oxide synthase activity. The effects of inhaled nitric oxide on the downstream mediators of the nitric oxide/cyclic guanosine monophosphate cascade, soluble guanylate cyclase and phosphodiesterase 5, have not been investigated. We sought to determine the effects of inhaled nitric oxide on endogenous cyclic guanosine monophosphate levels, soluble guanylate cyclase, and phosphodiesterase 5 protein levels in the intact lamb. METHODS: Eleven 1-month-old lambs were mechanically ventilated. In 7 lambs, inhaled nitric oxide (40 ppm) was administered for 24 hours and then acutely withdrawn. Intermittent lung biopsy samples were obtained for cyclic guanosine monophosphate concentrations and soluble guanylate cyclase and phosphodiesterase 5 protein levels (Western blot analysis). RESULTS: Initiation of nitric oxide decreased left pulmonary vascular resistance by 26.2%, and withdrawal rapidly increased pulmonary vascular resistance by 77.8% (P <.05). Tissue cyclic guanosine monophosphate concentrations initially increased during nitric oxide therapy but were not maintained during the 24-hour exposure. In addition, cyclic guanosine monophosphate concentrations rapidly decreased after nitric oxide withdrawal (P <.05). The alpha soluble guanylate cyclase (-45.7%) and beta soluble guanylate cyclase (-48.4%) protein levels decreased during nitric oxide therapy (P <.05), whereas phosphodiesterase 5 proteins levels were unchanged. CONCLUSIONS: These data suggest a role for decreased soluble guanylate cyclase and its resulting decrease in cyclic guanosine monophosphate concentrations in the nonsustained response to nitric oxide and the rebound pulmonary hypertension noted on its acute withdrawal. Phosphodiesterase 5 inhibitors may be a useful adjunct therapy during inhaled nitric oxide to preserve cyclic guanosine monophosphate levels and thereby preserve nitric oxide responsiveness and prevent rebound pulmonary hypertension. PMID- 15115985 TI - Xenoreactivity and engraftment of human mesenchymal stem cells transplanted into infarcted rat myocardium. AB - OBJECTIVE: It is thought that adult human mesenchymal stem cells do not induce immunoreactivity even to xenografts. We wanted to study whether adult human mesenchymal stem cells survive and engraft in experimentally induced ischemic rat myocardium. METHODS: Bone marrow-derived adult human mesenchymal stem cells (2.5 x 10(6)) were injected into the myocardium of 4 Sprague-Dawley rats. One week after injection, peripheral blood rat lymphocytes were added to adult human mesenchymal stem cells in a mixed lymphocyte reaction. Furthermore, an infarction was created by left anterior descending artery ligation of 8 Sprague-Dawley rats, 4 of which were immunosuppressed with tacrolimus (0.1 mg/kg/d) and 4 RNU athymic rats. One week after left anterior descending artery ligation, 2.5 to 3.5 x 10(6) adult human mesenchymal stem cells were injected around the infarcted area. The adult human mesenchymal stem cells were identified with fluorescence in situ hybridization technique and myocardial antigens by immunohistochemistry. The immune response was studied by hematoxylin and eosin staining and by antibodies directed toward macrophages. RESULTS: Significant rat lymphocyte proliferation was observed when adult human mesenchymal stem cells were added to peripheral blood from Sprague-Dawley rats previously exposed to adult human mesenchymal stem cells. No reactivity was seen in lymphocytes from untreated Sprague-Dawley rats and athymic rats. Adult human mesenchymal stem cells could only be identified in the myocardium of athymic rats. Further, in normal Sprague-Dawley rats, there was a significant myocardial infiltration of round cells, mostly macrophages, in the area of injection of adult human mesenchymal stem cells. In RNU rats, this reaction was less intense. CONCLUSION: Adult human mesenchymal stem cells did not induce xenoreactivity in vitro in previously unexposed immunocompetent Sprague Dawley rats. However, although mesenchymal stem cells are transplantable across allogeneic barriers, transplant rejection can occur in a xenogenic model. When transplanted into an immunoincompetent host, adult human mesenchymal stem cells showed persistent engraftment. PMID- 15115986 TI - Hypoxia: unique myocardial morphology? AB - OBJECTIVE: The objective of this study was to investigate the effects of chronic and intermittent hypoxia on myocardial morphology. METHODS: Rats randomly divided into 3 groups (n = 14 per group) were exposed to room air (Fio(2) = 0.21), chronic hypoxia (Fio(2) = 0.10), and intermittent hypoxia (chronic hypoxia with 1 hour per day of room air) for 2 weeks. Weight, blood gas analysis, hematocrit, hemoglobin, red cells, and right and left ventricular pressures were measured. Hearts excised for morphologic examination were randomly divided into 2 groups (9 per group for gross morphologic measurements and 5 per group for histologic and morphometric analysis). The weight ratio of right to left ventricles plus interventricular septum, myocyte diameter, cross-sectional area, and free wall thickness in right and left ventricles were measured. RESULTS: Despite the same polycythemia, the right ventricle pressure (P <.05) and ratio of right to left ventricle pressures (P <.02) were higher after chronic hypoxia than intermittent hypoxia. The ratio of heart weight to total body weight and the ratio of right to left ventricles plus interventricular septum was higher (P <.01) in chronic and intermittent hypoxia than in normoxia. Myocyte diameter was not different between the right and left ventricles in normoxia, whereas right ventricle myocytes were larger than left ventricle myocytes in chronic hypoxia (P <.05) and intermittent hypoxia (P <.0005). There was marked dilatation of right ventricle size (P <.001) and marked reduction of left ventricle (P <.001) size in chronic and intermittent hypoxia compared with normoxia. The total ventricular area (right ventricle plus left ventricle area) remained the same in all groups. The wall thickness ratio in chronic hypoxia and intermittent hypoxia was increased (P <.001) compared with normoxia in the right ventricle but not in the left ventricle. CONCLUSIONS: Intermittent reoxygenation episodes do not induce a lesser ventricular hypertrophic response than observed with chronic hypoxia. The functional myocardial preconditioning consequence of intermittent reoxygenation is not supported by structural differences evident with the available techniques. PMID- 15115987 TI - Bridging to transplant with the HeartMate left ventricular assist device: The Columbia Presbyterian 12-year experience. AB - OBJECTIVE: Implantation of a left ventricular assist device as a bridge to transplantation has become an acceptable approach for patients with end-stage heart failure. Our long-term results with 3 Thoratec HeartMate devices are presented to outline improvements in successful bridging to transplantation and post-transplant survival. METHODS: From August 1990 through January 2003, 243 patients underwent implantation of Thoratec HeartMate devices as a bridge to transplantation. This included 52 (21.4%) pneumatic devices, 17 (7.0%) dual-lead vented electric devices, and 174 (71.6%) single-lead vented electric devices. RESULTS: Mean age was 49.7 +/- 13.7 years. Mean support time was 78.1 +/- 82.9 days (0-541). Bridging success increased from 63.5% (n = 33) for pneumatic devices to 64.7% (n = 11) for dual-lead vented electric devices and 72.4% (n = 126) for single-lead vented electric devices (P =.005). Posttransplant 1-, 3-, and 5-year actuarial survival increased from 87.5%, 78.1%, and 71.9% in patients with pneumatic devices to 91.5%, 86.9%, and 81.3%, respectively, for patients with single-lead vented electric devices. Device infection and malfunction occurred in 17.7% (n = 43) and 12.8% (n = 31) of patients, respectively. CONCLUSIONS: Successful bridging to transplantation and posttransplant survival has improved over time. Left ventricular assist devices have become increasingly more effective in bridging patients with end-stage heart failure to transplantation. This is likely due to a combination of better patient selection, improvements in clinical practice, and evolution in device design. PMID- 15115988 TI - The endothelin 1A receptor antagonist BSF 302146 is a potent inhibitor of neointimal and medial thickening in porcine saphenous vein-carotid artery interposition grafts. AB - OBJECTIVE: Late saphenous vein graft failure after coronary artery bypass graft surgery is initiated by medial thickening and neointima formation, both of which are mediated by the proliferation of vascular smooth muscle cells. Because porcine vein grafts contain high levels of endothelin 1 receptor subtypes and endothelin 1 promotes the proliferation of vascular smooth muscle cells, the effect of administration of the endothelin 1(A) receptor antagonist BSF 302146 ([+]-[S]-2-[4,6-dimethyl-pyrimidin-2-yloxy]-3,3-diphenyl-butanoic acid) on porcine vein graft thickening was investigated. METHODS: Saphenous vein-carotid artery interposition grafting was performed in 4 groups of large white pigs (30 35 kg, n = 10 for each group). BSF 302146 was administered orally (3, 10, and 30 mg x kg(-1) x d(-1)) for 4 weeks to one group of pigs, and placebo was administered to the other group (control animals). Pigs were then anesthetized, and the grafts were removed and fixed at 100 mm Hg with 4% paraformaldehyde. Histologic sections were prepared, and graft morphometry was carried out by using computer-aided planimetry. RESULTS: In vein grafts from animals treated with BSF 302146 compared with grafts from control animals (untreated), there were significant dose-dependent reductions in the increase in medial thickness and neointimal thickness, an increase in luminal area, and a decrease in proliferating cell nuclear antigen-positive cells in the medial-intimal area. CONCLUSIONS: The administration of BSF 302146 reduces graft thickening and promotes positive remodeling through an endothelin 1(A)-mediated effect on vascular smooth muscle cell replication. The administration of this endothelin 1(A) receptor antagonist might therefore be therapeutically effective in preventing late vein graft failure in patients undergoing coronary artery bypass grafting. PMID- 15115989 TI - Lung cancer resection combined with lung volume reduction in patients with severe emphysema. AB - OBJECTIVE: Certain patients with resectable lung cancer and severe respiratory limitation due to emphysema may have a suitable operative risk by combining cancer resection with lung volume reduction surgery. The purpose of this study is to review our experience with such patients. METHODS: A review was conducted on 21 patients with lung cancer in the setting of severe emphysema who underwent an operation designed to provide complete cancer resection and volume reduction effect. RESULTS: In the 21 patients, the mean preoperative forced expiratory volume in 1 second was 0.7 +/- 0.2 L (29% predicted), residual volume was 5.5 +/- 1.0 L (271%), and diffusing capacity for carbon monoxide was 8.0 +/- 2.2 mL/min/mm Hg (34% predicted). In 9 patients, the cancer was located in a severely emphysematous lobe and the lung volume reduction surgery component of the procedure was accomplished with lobectomy alone. In the remaining 12 patients, the cancer resection lobectomy (n = 9) and wedge resection (n = 3) were supplemented with lung volume reduction surgery. Final pathologic staging was stage I in 16 patients, stage II in 2 patients, and stage III in 2 patients. One patient was found to have stage IV disease due to multifocal tumors in separate lobes. There were no hospital deaths. Postoperative complications included prolonged air leak in 11 patients, atrial fibrillation in 6 patients, and reintubation for ventilatory assistance in 2 patients. All patients showed improved lung function postoperatively. Survival was 100% and 62.7% at 1 and 5 years, respectively. CONCLUSIONS: Patients with severe emphysema and resectable lung cancer who have a favorable anatomy for lung volume reduction surgery may undergo a combined cancer resection and lung volume reduction surgery with an acceptable risk and good long-term survival. PMID- 15115990 TI - Expression profiling of non-small cell lung carcinoma identifies metastatic genotypes based on lymph node tumor burden. AB - OBJECTIVE: This study hypothesized that non-small cell lung carcinoma cells from primary tumors isolated by laser capture microdissection would exhibit gene expression profiles associated with graded lymph node metastatic cell burden. METHODS: Non-small cell lung carcinoma tumors (n = 15) were classified on the basis of nodal metastatic cell burden by 2 methods, obtaining 3 groups: no metastasis, micrometastasis, and overt metastasis. We then performed microarray analysis on microdissected primary tumor cells and identified gene expression profiles associated with graded nodal tumor burden using a correlation-based selection algorithm coupled with cross-validation analysis. Hierarchical clustering showed the regrouping of tumor specimens; the classification inference was assessed with Fisher's exact test. We verified data for certain genes by using another independent assay. RESULTS: The 15 specimens clustered into 3 groups: cluster A predominated in specimens with overt nodal metastasis; cluster B had more specimens with nodal micrometastases; and cluster C included only specimens without nodal metastases. Cluster assignment was based on a validated 75-gene discriminatory subset. Notably, genes not previously associated with positive non-small cell lung carcinoma lymph node status were encountered in the profiling analysis. CONCLUSIONS: Microdissection, combined with microarray analysis, is a potentially powerful method to characterize the molecular profile of tumor cells. The 75-gene expression profiles representative of clusters A and B may define genotypes prone to metastasize. Overall, the 3 groups of tumor specimens clustered separately, suggesting that this approach may identify graded metastatic propensity. Further, genes singled out in clustering may yield insights into underlying metastatic mechanisms and may represent new therapeutic targets. PMID- 15115991 TI - Experimental study of extracorporeal lung resection in dogs: ex situ sleeve resection and autotransplantation of the pulmonary lobe after extended pneumonectomy for central lung cancer. AB - OBJECTIVE: Extracorporeal lung resection as an alternative to pneumonectomy for central lung cancer is a procedure in which the unilateral lung is extirpated, removing the pulmonary lobe with the cancers and replanting the residual pulmonary lobe. The aim of this study was to investigate whether extracorporeal lung resection for lung cancer can be performed safely. METHODS: Nineteen dogs were divided into the control and extracorporeal lung resection groups. The former (n = 5) underwent lung autotransplantion, and the latter was subdivided into ND1 (n = 7) and ND2 (n = 7) groups on the basis of the manner of lymph node dissection. By comparing the 3 groups, the adverse effects of lymph node dissection were examined. RESULTS: All dogs in the control group had no complications. Four dogs in the ND1 group survived for 90 to 630 days after the operation. In the ND2 group 5 dogs succumbed within 30 days after the operation, although the other 2 dogs survived for 391 and 573 days, respectively. Bronchopulmonary fistulas were seen in 1 of the ND1 dogs and 3 of the ND2 dogs. Two of the latter were free of thrombus formation in the pulmonary arteries and veins of the autografts. In the ND2 group, compared with the control and ND1 group, the tissue blood flow at the bronchial anastomotic site indicated reduction between the 3rd and 14th postoperative days. CONCLUSION: The extensive lymph node dissection had severe adverse effects on bronchial anastomotic healing in extracorporeal lung resection. Therefore extracorporeal lung resection can be applied to only a very limited number of patients with N0 or N1 disease. PMID- 15115992 TI - Safety and efficacy of median sternotomy versus video-assisted thoracic surgery for lung volume reduction surgery. AB - BACKGROUND: The National Emphysema Treatment Trial, a randomized trial comparing lung volume reduction surgery with medical therapy for severe emphysema, included randomized and nonrandomized comparisons of the median sternotomy and video assisted thoracoscopic approaches for lung volume reduction surgery. METHODS: Lung volume reduction surgery was performed by median sternotomy only at 8 centers and video-assisted thoracoscopy only at 3 centers; 6 centers randomized the approach to lung volume reduction surgery. Mortality, morbidity, functional status, and costs were assessed. RESULTS: In the nonrandomized comparison, 359 patients received lung volume reduction surgery by median sternotomy, and 152 patients received lung volume reduction surgery by video-assisted thoracoscopy. The 90-day mortality was 5.9% for median sternotomy and 4.6% for video-assisted thoracoscopy (P =.67). Overall mortality was 0.08 deaths per person-year for median sternotomy and 0.10 deaths per person-year for video-assisted thoracoscopy (video-assisted thoracoscopy-median sternotomy risk ratio, 1.18; P =.42). Complication rates were low and not statistically different for the 2 approaches. The median hospital length of stay was longer for median sternotomy than for video-assisted thoracoscopy (10 vs 9 days; P =.01). By 30 days after surgery, 70.5% of median sternotomy patients and 80.9% of video-assisted thoracoscopy patients were living independently (P =.02). Functional outcomes were similar for median sternotomy and video-assisted thoracoscopy at 12 and 24 months. Costs for the operation and the associated hospital stay and costs in the 6 months after surgery were both less for video-assisted thoracoscopy than for median sternotomy (P <.01 in both cases). Similar results were noted for the randomized comparison. CONCLUSIONS: Morbidity and mortality were comparable after lung volume reduction surgery by video-assisted thoracoscopy or median sternotomy, as were functional results. The video-assisted thoracoscopic approach to lung volume reduction surgery allowed earlier recovery at a lower cost than median sternotomy. PMID- 15115993 TI - Surgical management of childhood bronchiectasis due to infectious disease. AB - BACKGROUND: The purpose of this study was to estimate operative risk and to identify indicators of adverse prognosis in patients undergoing resection for childhood bronchiectasis. METHODS: From January 1985 to February 2001, patients undergoing resection for bronchiectasis were studied. The indications for operation were failure of medical therapy in 33 patients (94.2%) and hemoptysis in 2 (5.7%). The mean duration of symptoms was 4.2 years (range, 1-9 years). Surgical treatment included lobectomy in 17 patients (48.5%), pneumonectomy in 7 (20%), lobectomy plus segmentectomy in 5 (14.2%), bilobectomy in 2 (5.7%), and segmentectomy in 4 (11.4%). RESULTS: The operative mortality rate was 2.8%, and the morbidity rate was 17.6%. The mean follow-up in 34 patients was 5.4 years (range, 1-12 years). Overall, 22 patients (64.7%) were asymptomatic after surgery. Clinical improvement was noticed in 8 patients (23.5%), and no improvement was noticed in 4 (11.7%). Complete resection resulted in a significantly better clinical outcome than incomplete resection (P <.05). CONCLUSIONS: Surgery for childhood bronchiectasis can be performed with low mortality and morbidity. Complete resection should be performed when possible. PMID- 15115994 TI - Cost-effectiveness of pulmonary resection and systemic chemotherapy in the management of metastatic soft tissue sarcoma: a combined analysis from the University of Texas M. D. Anderson and Memorial Sloan-Kettering Cancer Centers. AB - BACKGROUND: We sought to determine the cost-effectiveness of different treatment strategies for patients with pulmonary metastases from soft tissue sarcoma. METHODS: We constructed a decision tree to model the outcomes of 4 treatment strategies for patients with pulmonary metastases from soft tissue sarcoma: pulmonary resection, systemic chemotherapy, pulmonary resection and systemic chemotherapy, and no treatment. Data from 1124 patients with pulmonary metastases from soft tissue sarcoma were used to estimate disease-specific survival for pulmonary resection and no treatment. Outcomes of systemic chemotherapy and pulmonary resection and of systemic chemotherapy were estimated by assuming a 12 month improvement in disease-specific survival with chemotherapy; this was done on the basis of the widely held but unproven assumption that chemotherapy provides a survival benefit in patients with metastatic soft tissue sarcoma. Direct costs were examined for a series of patients who underwent protocol-based pulmonary resection or doxorubicin/ifosfamide-based chemotherapy. RESULTS: The mean cost of pulmonary resection was 20,339 dollars per patient; the mean cost of 6 cycles of chemotherapy was 99,033 dollars. Compared with no treatment and assuming a 12-month survival advantage with chemotherapy, the incremental cost effectiveness ratio was 14,357 dollars per life-year gained for pulmonary resection, 104,210 dollars per life-year gained for systemic chemotherapy, and 51,159 dollars per life-year gained for pulmonary resection and systemic chemotherapy. Compared with pulmonary resection, the incremental cost effectiveness ratio of pulmonary resection and systemic chemotherapy was 108,036 dollars per life-year gained. Sensitivity analyses showed that certain patient and tumor features, as well as the assumed benefit of chemotherapy, affected cost effectiveness. CONCLUSIONS: For patients with pulmonary metastases from soft tissue sarcoma who were surgical candidates, pulmonary resection was the most cost-effective treatment strategy evaluated. Even with favorable assumptions regarding its clinical benefit, systemic chemotherapy alone, compared with no treatment, was not a cost-effective treatment strategy for these patients. PMID- 15115995 TI - Should the transverse aortic arch be replaced simultaneously with aortic root replacement for annuloaortic ectasia in Marfan syndrome? AB - OBJECTIVE: The purpose of this study was to determine, on the basis of the late fate of the intact aortic arch with abnormal tissue after aortic root replacement, whether the intact aortic arch should be replaced prophylactically at the time of aortic root replacement for annuloaortic ectasia in Marfan syndrome. METHODS: A retrospective review was performed in 85 patients with Marfan syndrome who underwent aortic root replacement for annuloaortic ectasia with or without aortic dissection (mean age 37 years, range 19-61 years). These 85 patients were divided into four groups according to the postoperative condition of the residual aorta. In group I (n = 47), the patients underwent aortic root replacement for annuloaortic ectasia with or without localized dissection in the ascending aorta. In these patients the residual aorta, including the aortic arch, was therefore intact. In group II (n = 10), the aortic arch was intact, although the descending thoracic aorta was dissected because of the preoperative type B dissection. In groups III and IV, the patients had type A dissection involving the transverse arch associated with annuloaortic ectasia. In group III (n = 13), residual dissection existed in the descending thoracic aorta after concomitant total arch replacement. In group IV (n = 15), the aortic arch and the descending thoracic aorta were dissected. RESULTS: There were 5 early deaths (3 in group I, 1 in group II, and 1 in group III). Subsequent operations were required in 10, 5, 6, and 7 cases in groups I, II, III, and IV, respectively. Regarding the aortic arch, only 2 of 53 survivors of the initial hospitalization with an intact aortic arch (groups I and II) underwent subsequent total arch replacement for the onset of dissection in the aortic arch, and 4 of 14 survivors of the initial hospitalization with a residual dissecting arch (group III) needed subsequent total arch replacement. Actuarial freedom from arch repair among patients with an intact aortic arch (91% at 15 years) was significantly higher than that among patients with a residual dissecting arch (49% at 15 years, P =.0078). CONCLUSIONS: The incidence of new dissection in the residual intact arch after aortic root replacement was extremely low. Therefore prophylactic replacement of the intact arch does not appear to be necessary at aortic root replacement for annuloaortic ectasia in Marfan syndrome. PMID- 15115996 TI - Long-term durability of resection and end-to-end anastomosis for ascending aortic aneurysms. AB - BACKGROUND: Ascending aortic aneurysms with normal sized sinotubular junction are generally treated by resection of the dilated aorta and replacement with tubular graft. Aortic resection and direct end-to-end anastomosis has been applied to repair aortic coarctation, interrupted aortic arch, and traumatic aortic rupture. No data exist regarding the long-term durability of this approach in ascending aortic aneurysms. The aim of this case-control study was to illustrate the durability of this operation by presenting our entire experience and the long term follow up of a cohort of 34 patients who underwent ascending aortic aneurysm resection and primary end-to-end anastomosis between January 1990 and March 2003 in Caen University Hospital (Caen, France). METHODS: The mean age of patients was 61.5 +/- 12.5 years, and there were 18 male and 16 female patients. The operative technique included extensive mobilization of the arch, supra-aortic trunks, and inferior vena cava to enable approximation of the aortic ends, thus avoiding tension on the suture lines. Associated aortic valve replacement was performed in 27 patients; mechanical valves were used in 19. A bicuspid aortic valve was present in 9 patients; in 3 cases the valve was regurgitant. Aortic valve regurgitation was present in a total of 7 patients. Patients were followed up at regular intervals; total follow-up was 2187 patient-months, with a median follow up time of 72 months per patient (25th-75th percentile 10.5-102.7 months). RESULTS: One patient died 10 days after the operation of aortic rupture related to suture infection caused by mediastinitis. Late deaths occurred in 3 patients, who died 12, 62, and 71 months after the operation, but none of these deaths were attributable to late aortic repair failure. No patient in this series required reoperation, including patients with aortic regurgitation or bicuspid aortic valve. Follow-up was 91.1% complete at the closing date of April 1, 2003. The Kaplan-Meier estimate of survival for all patients was 120.4 months (95% confidence interval 105.1-135.7 months). The median of preoperative maximal aortic diameter was 55.1 mm (range 50.3 to 67.5 mm, 25th-75th percentile 50.5 56.8 mm). The median immediate postoperative diameter was 40.3 mm (range 33.4 46.4 mm, 25th-75th percentile 37.2-42.0 mm, P <.0001 relative to preoperative diameter), and the median length of the resected aortic segment was 52 mm (range 48-76 mm, 25th-75th percentile 50.1-66.4 mm). The median decrease of aortic diameter was 24.9 mm (range 8.9-32.6 mm, 25th-75th percentile 18.2-26.6 mm). The median aortic diameter at the end of the follow-up was 41.0 mm (range 34.6-46.1 mm, 25th-75th percentile 37.0-43.2 mm, P =.6 relative to immediate postoperative diameter). CONCLUSIONS: Ascending aorta aneurysm resection and primary end-to-end anastomosis provides effective long-term outcome and in selected cases represents a good alternative to aortic interposition grafting. Aortic regurgitation and bicuspid aortic valve do not represent a contraindication for this treatment. PMID- 15115997 TI - Prosthetic valve thrombosis: twenty-year experience at the Montreal Heart Institute. AB - BACKGROUND: Prosthetic valve thrombosis is a life-threatening complication. We reviewed the incidence, risk factors, and treatment strategies of this rare complication. METHODS: From February 1981 through January 2001, 5430 valve operations were performed in 4924 patients at the Montreal Heart Institute. Of this cohort, 39 patients presented with prosthetic valve thrombosis and had complete follow-up data obtained from our prospective valve clinic database. RESULTS: In this series 82% of patients were women, and the mean age was 58 +/- 11 years. The underlying pathology involved the mitral valve in 75% of cases. Most prosthetic valve thromboses occurred with mechanical prostheses (95%). The time interval from first valve replacement to prosthetic valve thrombosis was 39 +/- 42 months. The most frequent clinical presentation was severe congestive heart failure (44%). On prosthetic valve thrombosis presentation, the international normalized ratio was less than 2.5 in 54%, with inadequate anticoagulation management in 26% and poor compliance in 26%. Eighty-two percent of patients underwent a surgical procedure, consisting of thrombectomy in 47%, mitral valve replacement in 47%, and aortic valve replacement in 6% of patients. The 30-day operative mortality and total in-hospital mortality after prosthetic valve thrombosis were 25% and 41%, respectively. The 10-year actuarial survival after prosthetic valve thrombosis was 46% +/- 10%. CONCLUSION: Inadequate level of anticoagulation is the most important factor involved in the pathogenesis of prosthetic valve thrombosis. The overall mortality rate despite surgical treatment remains high. This study underscores the importance of meticulous surveillance of anticoagulation therapy in patients with prosthetic valves. PMID- 15115998 TI - Penetrating atherosclerotic ulcer of the descending thoracic aorta and arch. AB - OBJECTIVE: The clinical behavior of penetrating atherosclerotic ulcers of the aorta is controversial. We reviewed our experience with this entity over a 25 year interval. METHODS: Cases were identified using the Department of Radiology database searching for the diagnoses of aortic dissection, intramural hematoma, or penetrating ulcer between 1977 and 2002. Available imaging studies were reviewed by a vascular radiologist to confirm the diagnosis of penetrating ulcer and perform serial measurements. RESULTS: One hundred five patients with penetrating atherosclerotic ulcers of the descending thoracic aorta or arch with (n = 85) or without (n = 20) associated intramural hematoma were confirmed. Two patients with ulcers in the ascending aorta were excluded. There were 73 men and 32 women with a mean age of 72 +/- 9 years. Comorbidities included hypertension in 97 (92%), tobacco use in 81 (77%), and coronary artery disease in 48 (46%). Of nonoperated patients with follow-up studies, the mean thickness of the intramural hematoma decreased at 1 month in 89% and completely resolved at 1 year in 85%. There were 3 deaths (4%) within 30 days among 76 patients treated medically and 6 deaths (21%) among 29 patients treated surgically (P <.05). Failure of medical therapy defined as surgery or death was predicted by rupture at presentation (odds ratio = 20.6) and era of treatment (before 1990, odds ratio 9.9) but not aortic diameter, ulcer size, or extent of hematoma. CONCLUSION: Although careful follow-up is necessary, many penetrating atherosclerotic ulcers of the thoracic aorta can be managed nonoperatively in the acute setting. PMID- 15115999 TI - Early quantitative coronary angiography of saphenous vein grafts for coronary artery bypass grafting harvested by means of open versus endoscopic saphenectomy: a prospective randomized trial. AB - OBJECTIVE: Endoscopic saphenectomy is associated with a decreased incidence of wound complications without an increase in histologic trauma or endothelial dysfunction in published reports. Concern remains about the patency of saphenous vein grafts harvested endoscopically and the development of early intimal hyperplasia. The purpose of this study was to compare early quantitative coronary analysis of saphenous vein grafts used for coronary artery bypass grafting harvested with the open versus endoscopic techniques. METHODS: Forty patients undergoing primary coronary artery bypass grafting surgery with at least 1 saphenous vein graft were randomized preoperatively to open versus endoscopic saphenectomy with bipolar cauterization of side branches. Quantitative coronary angiography was performed a mean of 3 months (range, 1-9 months) after the operation. RESULTS: There was no statistically significant difference in the patency rates of internal thoracic artery grafts between the open and endoscopic groups and no statistically significant difference in the patency rates of saphenous vein grafts between both groups (85.2% vs 84.4%, P =.991). Quantitative coronary angiography showed no difference in graft stenosis (>or=50% of the internal diameter of the graft) in the body of the saphenous vein grafts in the open versus endoscopic saphenectomy groups (3.7% vs 0%, P =.280). CONCLUSION: Angiographic appearance and patency rates of saphenous vein grafts harvested with the endoscopic technique are similar to those of saphenous vein grafts harvested with the open technique. These results support the use of endoscopic saphenectomy because of the known lower incidence of wound and infectious complications and superior functional results. PMID- 15116000 TI - Single versus bilateral internal thoracic artery grafts with concomitant saphenous vein grafts for multivessel coronary artery bypass grafting: effects on mortality and event-free survival. AB - BACKGROUND: The issue of superiority of single internal thoracic artery grafting versus bilateral internal thoracic artery grafting remains unresolved. The aim of this study was to compare the long-term outcome of single and bilateral internal thoracic artery grafting with concomitant saphenous vein grafting for multivessel coronary artery bypass grafting. METHODS: Between March 1985 and April 1995, 6650 patients underwent primary isolated coronary artery bypass grafting with internal thoracic artery grafts, including 4382 patients with multivessel bypass grafting requiring at least 3 grafts. Outcomes of patients undergoing single internal thoracic artery plus saphenous vein grafting (n = 2547) and bilateral internal thoracic artery plus saphenous vein grafting (n = 1835) were obtained at a mean follow-up of 11 +/- 3 years. RESULTS: Patients with bilateral internal thoracic artery grafting were younger, were mostly male, and had less diabetes, hypertension, unstable angina, and recent myocardial infarction than patients undergoing single internal thoracic artery grafting. Thirty-day mortality was 2.3% for the group undergoing single internal thoracic artery grafting versus 1.2% for those undergoing bilateral internal thoracic artery grafting (P =.007). Survival probability at 10 years was 88% for the single-graft group compared with 93% for the bilateral-graft group (P <.001). Multivariate analysis with propensity scoring showed that bilateral internal thoracic artery grafting decreased the risk of death (hazard ratio, 0.74; 95% confidence interval, 0.60 0.90), myocardial infarction (hazard ratio, 0.79; 95% confidence interval, 0.67 0.93), and coronary reoperation (hazard ratio, 0.41; 95% confidence interval, 0.21-0.80) throughout the follow-up period. Other significant predictors of death were diabetes, prior myocardial infarction, need for intra-aortic balloon pump, chronic heart failure, and peripheral vascular disease. CONCLUSION: Patients undergoing bilateral internal thoracic plus saphenous vein grafting appear to have a significantly better long-term clinical outcome than patients undergoing single internal thoracic artery plus saphenous vein grafting for multivessel coronary artery bypass grafting. PMID- 15116001 TI - Replacement of the aortic root for acute prosthetic valve endocarditis: prosthetic composite versus aortic allograft root replacement. AB - OBJECTIVE: Aortic root replacement for prosthetic aortic valve endocarditis with accompanying destruction of the aortic root is a well-established surgical intervention. However, there is still no consensus whether prosthetic material or allogeneic material should be used. Here we report on our experience with prosthetic composite and aortic allograft root replacement in such patients during a 10-year interval. METHODS: From 1991 through 2001, 29 patients with prosthetic aortic valve endocarditis combined with aortic root destruction underwent reoperation at our institution. Sixteen patients received aortic root replacement with a cryopreserved aortic root allograft (group A) and 13 with a prosthetic composite graft (group B). The interval between the initial operation and reoperation was 29 months (range, 5-168 months) in group A and 55 months (range, 7-248 months) in group B. RESULTS: Hospital mortality was 18.5% (n = 5 patients, 3 in group A and 2 in group B). Median follow-up was 21 months (range, 1-48 months) for group A and 34 months (range, 1-152 months) for group B (P >.2). Survival at 1 and 5 years was 81% +/- 10% and 81% +/- 10% in group A and 85% +/- 10% and 85% +/- 10% in group B, respectively. No patient underwent reoperation for recurrent prosthetic aortic valve endocarditis. CONCLUSIONS: Our results indicate that excellent long-term results can be achieved regardless of the material used for aortic root replacement in patients with prosthetic aortic valve endocarditis. PMID- 15116002 TI - Early bypass occlusion after deployment of nitinol connector devices. AB - BACKGROUND: Reducing the negative side effects associated with extracorporeal circulation is the major advantage of off-pump revascularization. However, side clamping of a calcified aorta for proximal anastomoses can cause emboli, resulting in neurologic damage. This problem has been addressed by introducing a mechanical anastomosis device (Symmetry, St Jude Medical) that allows vein-to aorta anastomosis without manipulating the aorta. This report describes our experience with this device. METHODS: Between June 2001 and April 2002, 77 connectors (1.3 per patient) were deployed in 61 patients (51 men and 10 women; mean age, 68 +/- 8.6 years) undergoing off-pump coronary artery bypass grafting or beating-heart revascularization. Intraoperative quality assessment included transit-time flow measurement (Medistim) and indocyanine green-based angiography (Spy, Novadaq). RESULTS: The surgeons were meticulously trained in loading of the device. No postoperative neurologic deficits were detected. Fifty-three patients had an uneventful course. However, 8 (13.1%) patients with 12 implanted connectors were symptomatic within 8 months (1 day to 8 months). Angiography revealed significant (95%) stenosis or even occlusion of the proximal vein-to aorta anastomosis at the level of all connectors. Four patients underwent reoperation (2 dilated-stented and 2 treated with drugs). CONCLUSION: On the basis of these observations, the routine use of the connector was halted at our institution. At the moment, the use of this therapy is reserved for patients with severely calcified aortas with no technical alternative. Further investigations appear necessary to evaluate the clinical patterns of this otherwise promising technology. PMID- 15116003 TI - Cryorecanalization: a new approach for the immediate management of acute airway obstruction. AB - OBJECTIVES: Endobronchial cryotherapy is an established recanalization method for stenoses of the respiratory tract. However, previous applications of cryotherapy have not been immediately effective, requiring a second clean-up procedure several days later to finally obtain sufficient recanalization. In this study we demonstrate a newly developed cryoprobe allowing recanalization of tumor stenoses during a single intervention. METHODS: In this prospective study flexible bronchoscopy was used for cryorecanalization of 60 patients with high-grade stenoses of the respiratory tract from exophytic tumors. Tumor tissue was frozen on the tip of the probe and subsequently removed from the surrounding respiratory tract tissue through retraction of the probe. Procedures were carried out on sedated, spontaneously breathing, intubated patients. RESULTS: Fifty (83%) of 60 patients were successfully or partially successfully treated. Tumor bleeding occurred in 6 patients but was stopped with argon plasma coagulator treatment in all patients not requiring rigid bronchoscopy. CONCLUSION: Cryorecanalization with the newly developed cryoprobe permits effective, safe, and inexpensive therapy of endobronchial stenoses of the respiratory tract. PMID- 15116004 TI - Psychosocial and sexual concerns of patients with implantable left ventricular assist devices: a pilot study. AB - OBJECTIVE: We sought to determine the psychosocial and sexual concerns of patients discharged from the hospital with implantable left ventricular assist devices. METHODS: Bridge-to-transplant patients with the Heartmate left ventricular assist device received a psychosocial and sexual survey at 1 month after discharge from the hospital. The survey consisted of 3 parts, with 5 questions in each category. The patients were asked to complete the survey by circling the responses and to provide more detailed answers when necessary. RESULTS: There were 8 male patients who completed the questionnaire. Psychologically, all patients expressed a positive mood and found support from family or religious sources. The majority described a change in attitude and behavior. Socially, all patients described a change in lifestyle, as well as a change in the reaction of family and friends. No patient went back to work or had the desire to return to work until after transplantation. The minority of patients admitted to smoking or drinking. Sexually, the majority had the desire or participated in sexual activities. The majority of patients either used or were interested in using sexual stimulants. There were no mechanical problems with the left ventricular assist devices. CONCLUSIONS: Psychosocial and sexual conditions are altered in patients with heart failure who are discharged with implantable left ventricular assist devices. At 1 month's time interval, there is an improvement in mood, an adjustment in lifestyle, and a positive shift in relations with family and friends. There is sexual desire, with interest in sexual-enhancing medication. The most common concern is related to the pump, such as the durability of the device or damage to the components. PMID- 15116005 TI - Insulin-like growth factor 1 improves the relationship between systemic oxygen consumption and delivery in piglets after cardiopulmonary bypass. AB - OBJECTIVE: We sought to assess the effects of insulin-like growth factor 1 on the balance between systemic oxygen consumption and oxygen delivery after cardiopulmonary bypass in piglets. METHODS: Twelve piglets weighing 4.5 to 8.3 kg undergoing hypothermic (28 degrees C) cardiopulmonary bypass for 70 to 120 minutes with 40 minutes of aortic crossclamping were studied before and during the first 6 hours after cardiopulmonary bypass. Oxygen consumption was continuously measured by an indirect calorimeter, Deltatrac II MBM-200 Metabolic Monitor (Datex Division Instrumentarium, Helsinki, Finland). Oxygen delivery and cardiac output were calculated from oxygen consumption and the arterial and mixed venous oxygen contents sampled before and every 30 minutes after cardiopulmonary bypass. Oxygen extraction ratio was derived by the ratio of oxygen consumption to oxygen delivery. Arterial blood lactate was measured before and every 30 minutes after cardiopulmonary bypass. Six animals were randomly assigned to receive an intravenous infusion of insulinlike growth factor 1 at 1.2 mg/h from 1 to 6 hours after cardiopulmonary bypass; the remaining 6 served as a control group. RESULTS: Relative to the control group, intravenous infusion of insulin-like growth factor 1 significantly reduced oxygen consumption (P =.02) and increased cardiac output (P =.016) and oxygen delivery (P =.049) during the first 6 hours after surgery with hypothermic cardiopulmonary bypass. As a result, oxygen extraction was significantly decreased (P =.012). CONCLUSIONS: Intravenous infusion of insulin like growth factor 1 improved oxygen transport by reducing oxygen consumption as well as increasing cardiac output and oxygen delivery during the first 6 hours after cardiopulmonary bypass in piglets. This may have important clinical implications for the care of critically ill children after surgery with cardiopulmonary bypass. PMID- 15116006 TI - Pulmonary expression of the hepatocyte growth factor receptor c-Met shifts from medial to intimal layer after cavopulmonary anastomosis. AB - OBJECTIVE: Pulmonary arteriovenous malformations occur in up to 60% of patients after cavopulmonary anastomosis. We compared the effects of cavopulmonary anastomosis and pulmonary artery banding on lung gene expression in an ovine model to study the abnormal pulmonary vascular remodeling after the exclusion of inferior vena caval blood independent of reduced pulmonary blood flow. We previously demonstrated by contrast echocardiography that pulmonary arteriovenous malformations develop by 8 weeks after cavopulmonary anastomosis but not after pulmonary artery banding. Hepatocyte growth factor, a pleiotropic factor with morphogenic, mitogenic, and angiogenic activities, signals via its specific receptor c-Met to induce the antiapoptotic factor Bcl-2. In this study, we examined pulmonary artery expression of these factors and their potential role in pulmonary artery remodeling after cavopulmonary anastomosis and pulmonary artery banding. METHODS: Eighteen lambs aged 35 to 45 days were placed into 3 groups: cavopulmonary anastomosis, pulmonary artery banding, and control (n = 6/group). In the cavopulmonary anastomosis group, the superior vena cava was anastomosed to the right pulmonary artery in an end-to-end fashion. In the pulmonary artery banding group, the left pulmonary artery was banded to reduce blood flow to 20% of control. The control group had a simple right pulmonary artery clamp for 30 minutes. Lung was harvested for Western blot, reverse transcriptase-polymerase chain reaction, and immunostaining at 2 weeks (n = 3/group) and 5 weeks (n = 3/group) after surgery. RESULTS: The expression of c-Met mRNA after cavopulmonary anastomosis was increased by twofold compared with the control or pulmonary artery banding group. The total lung expression of c-Met by Western blot was also up regulated at 2 weeks (P <.05). However, total lung expression of hepatocyte growth factor and Bcl-2 by Western and reverse transcriptase-polymerase chain reaction was not different from the control and pulmonary artery banding groups at both 2 and 5 weeks after surgery. Immunohistochemical analysis revealed that c Met expression was localized to the intimal layer of the pulmonary artery in the cavopulmonary anastomosis, while its expression in the control and pulmonary artery banding lungs was localized to the medial layer. Localization of Bcl-2 on the intimal layer in lambs with cavopulmonary anastomosis followed the same pattern as c-Met. CONCLUSIONS: After cavopulmonary anastomosis, pulmonary artery expression of the hepatocyte growth factor receptor c-Met and one of its downstream effectors, Bcl-2, had increased in the intimal layer and decreased in the medial layer. Because the hepatocyte growth factor signaling promotes increased endothelial cell survival, it may have a role in pulmonary artery remodeling following cavopulmonary anastomosis. In addition, the change of c-Met expression in the medial layer after cavopulmonary anastomosis suggests a possible mechanism for the smooth muscle cell alteration related to abnormal angiogenesis. PMID- 15116007 TI - Hypoxic pulmonary vasoconstriction disappears in a rabbit model of cavopulmonary shunt. AB - BACKGROUND: Cavopulmonary shunt is widely known as an interim staging procedure in patients with single-ventricle physiology. However, the physiologic characteristics of the pulmonary arterial system after cavopulmonary shunt are not clearly understood. In this article, we developed a rabbit cavopulmonary shunt model and studied the morphologic changes and physiologic characteristics (namely, hypoxic pulmonary vasoconstriction) of pulmonary arteries after cavopulmonary shunt. METHODS: Male Japanese white rabbits aged 12 to 16 weeks were used for the study. In 5 rabbits, the superior vena cava was anastomosed to the right pulmonary artery in an end-to-side fashion, followed by a proximal side ligation of the right pulmonary artery (cavopulmonary shunt group). In 4 rabbits, the superior vena cava and the right pulmonary artery were dissected and clamped for 10 minutes without making a cavopulmonary shunt (sham group). Two weeks after the operation, we then measured the internal diameter of the acinar (internal diameter, 164 +/- 7 microm), the lobular (305 +/- 13 microm), and the segmental (669 +/- 16 microm) pulmonary arteries in both controlled and hypoxic conditions by using a specially designed x-ray television system. Also, morphometric measurements were made in the pulmonary arteries around the terminal bronchioles. RESULTS: Two weeks after the operation, the arterial oxygen tension under room air conditions was significantly lower in the cavopulmonary shunt group than in the sham group (68.2 +/- 2.2 mm Hg vs 91.1 +/- 1.9 mm Hg; P =.01). The baseline internal diameters in the acinar and the lobular (resistance), but not the segmental (conduit), pulmonary arteries on the anastomosed side of the cavopulmonary shunt group were significantly larger than those of pulmonary arteries on the nonanastomosed side of the cavopulmonary shunt group and the sham group. Moreover, the pulmonary arteries on the anastomosed side of the cavopulmonary shunt group did not respond to hypoxia, whereas those on the nonanastomosed side of the cavopulmonary shunt and sham groups did have local internal diameter reductions in the acinar and lobular arteries (-1.1% +/- 1.0% in the anastomosed side vs -17.7% +/- 3.5% in the nonanastomosed side vs -20.9% +/- 6.1% in the sham group; P =.03). In the morphometric studies, the internal diameter of the pulmonary artery accompanying the terminal bronchiole in the anastomosed side of the cavopulmonary shunt group was significantly larger, and the ratio of medial thickness relative to the outer diameter was smaller compared with ratios in the nonanastomosed side of the cavopulmonary shunt group and the sham group. CONCLUSIONS: We developed a rabbit cavopulmonary shunt model. In the anastomosed side of the cavopulmonary shunt group, the peripheral pulmonary arteries, which contributed greatly in regulating the pulmonary vascular resistance, had a local reduction in the basal vascular tone and no hypoxic vasoconstriction 2 weeks after the operation. PMID- 15116008 TI - Phosphorylcholine or heparin coating for pediatric extracorporeal circulation causes similar biologic effects in neonates and infants. AB - OBJECTIVE: Cardiac surgery for complex congenital malformations with use of extracorporeal circulation predisposes to an excessive systemic inflammatory response and a consecutive capillary leak syndrome. In a prospective randomized study the influence of 2 oxygenators especially designed for pediatric use on inflammatory markers and clinical outcome was investigated. METHODS: Forty neonates and infants (body surface area, <0.36 m(2)) undergoing cardiac surgery with extracorporeal circulation were randomized into one of 3 groups: in the first group (n = 14) the Medtronic Minimax Oxygenator and in the second group (n = 12) the Dideco Lilliput 1 Oxygenator, both with a 750-mL priming volume, were used. In the third group the Dideco Lilliput 1 Oxygenator was filled with a reduced priming volume of 450 mL. Parameters of interest for evaluation of a systemic inflammatory response after extracorporeal circulation were interleukin 6, tumor necrosis factor alpha, neutrophil elastase, complement C3, and free hemoglobin. In addition, erythrocyte, leukocyte, and thrombocyte counts and hemoglobin and C-reactive protein values were determined at different measurement points before, during, and after the operation. RESULTS: In all 3 groups peak values for tumor necrosis factor alpha were observed during the operation, whereas interleukin 6, elastase, and free hemoglobin values peaked in the first 4 hours. The highest values for leukocytes and C-reactive protein were obtained between 24 and 72 hours after the operation. Erythrocyte and thrombocyte counts, as well as hemoglobin values, were lowest at extracorporeal circulation onset, normalizing under substitution in the first 4 hours after the operation. By using the Lilliput/750 oxygenator, higher interleukin 6 values 1 and 4 hours after the operation and higher tumor necrosis factor alpha values during and 1 hour after the operation could be observed compared with results with the Minimax and Lilliput/450 oxygenators. In spite of our randomization protocol, patients in the Lilliput/750 group were significantly smaller and younger than those in the Minimax group. However, the statistical analysis showed no correlation between age and interleukin 6 or tumor necrosis factor alpha values, but it did show a correlation between younger age and the occurrence of capillary leak syndrome. Accordingly, the number of children with clinically complicated course (capillary leak, longer duration of catecholamine therapy, and ventilation) was higher in the Lilliput/750 group than in the Minimax group. CONCLUSION: By using an adequate priming volume, the systemic inflammatory response is similar after use of the Dideco Lilliput 1 Oxygenator and the Medtronic Minimax Oxygenator. Tip-to tip surface coating of the extracorporeal circulation with either heparin or phosphorylcholine seems to have similar biologic effects in neonates and infants undergoing cardiac surgery. PMID- 15116009 TI - Management of infants with large, unrepaired ventricular septal defects and respiratory infection requiring mechanical ventilation. AB - OBJECTIVES: We sought to describe the hospital management and early outcome of critically ill infants presenting with large ventricular septal defects and pneumonia requiring mechanical ventilation at a referral center in a developing country. Infants with large ventricular septal defects who have pneumonia might present with respiratory failure requiring mechanical ventilation. In the developing world this presentation is relatively common, but few data exist describing patient management strategies. METHODS: Hospital data of consecutive infants admitted with large ventricular septal defects and pneumonia requiring mechanical ventilation were reviewed and analyzed. RESULTS: We identified 18 infants (mean age, 3.6 +/- 3.0 months). On admission, all the infants were significantly malnourished, and echocardiography showed bidirectional shunting (predominantly right-to-left shunting) in 6 infants. Thirteen (72%) patients improved with intensive medical management that included mechanical ventilation for 1 to 16 days (median, 6.5 days); unequivocal left-to-right shunting was subsequently documented by means of echocardiography in all 13 patients. Twelve patients underwent surgical repair, and 11 (91.6%) were discharged after median mechanical ventilation of 100 hours (range, 42-240 hours) and intensive care unit stay of 8 days (range, 4-15 days). Five of 6 unoperated patients died, 4 of them within a few hours of admission. One child with multiple ventricular septal defects was discharged and subsequently underwent pulmonary artery banding. CONCLUSION: Corrective cardiac surgery for selected critically ill infants with large ventricular septal defects, severe malnutrition, and pneumonia requiring mechanical ventilation is feasible and should be considered a viable management strategy. PMID- 15116010 TI - The influence of right anterolateral thoracotomy in prepubescent female patients on late breast development and on the incidence of scoliosis. AB - BACKGROUND: It is assumed that a right anterolateral thoracotomy for correction of simple congenital cardiac defects (ie, atrial septal defect) achieves more favorable cosmetic results than a standard median sternotomy. METHODS: Ninety five patients, 72 with right anterolateral thoracotomy and 23 with median sternotomy, who had corrective transatrial operations when they were younger than 12 years of age were contacted by questionnaire. The mean follow-up time was 23.1 years. Of these, 61 patients (46 thoracotomy and 15 sternotomy) were investigated clinically. Volume differences of the breasts were measured by 3-dimensional surface scanning. By using photographs of the upper chest, breast symmetry was described by an index. The degree of scoliosis was measured by clinical examination. RESULTS: According to the questionnaire analysis, 76% (thoracotomy group) versus 39% (sternotomy group) thought that the cosmetic result was excellent (P =.008). Breast volume measurement showed a volume difference greater than 20% (left side larger than right) in 55% (thoracotomy) versus 0% (sternotomy). With our index, asymmetry in the lower part of the right breast occurred in 61% (thoracotomy) versus 0% (sternotomy; P <.001). A total of 6.6% of the patients had scoliosis, without any differences between groups. CONCLUSIONS: Because our long-term follow-up in prepubescent female patients after right anterolateral thoracotomy revealed significantly impaired unilateral breast development, we propose to abandon right anterolateral thoracotomy in this subgroup of patients, although the subjective satisfaction with the cosmetic result was high. To avoid potential damage of future breast tissue, other surgical approaches, such as right posterior thoracotomy, should be considered. According to the orthopedic investigation, the surgical approach does not cause a higher rate of scoliosis. PMID- 15116011 TI - Survival of patients removed from the heart transplant waiting list. AB - OBJECTIVE: End-stage heart failure has been associated with high mortality in the absence of transplantation. We evaluated the outcome of patients receiving optimal medical therapy who were removed from the cardiac transplant waiting list to determine survival and predictors of mortality. METHODS: We performed a retrospective review of 27 patients removed from the cardiac transplant waiting list from 1999 to 2001 at our institution. RESULTS: Mean age was 53 +/- 11 years; 16 of the patients were male. Status was IB in 3 cases and II in 24. Median time on the list was 32 months, and median follow-up was 2.9 years. Patients were removed from the transplant list because of either clinical improvement (group A, n = 18) or deterioration (group B, n = 9). In group A, 13 patients had improved functional status and 10 were in New York Heart Association class 1 or 2; 16 had improved echocardiographic left ventricular function. Survivals at 3 years were 100% in group A and 44% in group B (P <.01). CONCLUSION: Patients with end-stage heart failure who have clinical response to medical therapy have excellent 3-year survival. These data suggest the necessity of close evaluation of patients waiting for transplantation, with a low threshold for inactivation if persistent clinical improvement is observed. PMID- 15116012 TI - Heart transplantation in diabetic recipients: a decade review of 161 patients at Columbia Presbyterian. AB - OBJECTIVE: Diabetes is considered by some transplant centers to be a relative contraindication for cardiac transplantation because of concerns regarding decreased survival, as well as increased incidence of infection and transplant coronary artery disease. We evaluated our experience with diabetic recipients over the last 10 years. METHODS: From January 1992 through June 2002, 881 patients underwent cardiac transplantation at New York Presbyterian Hospital. Of these, 161 (18.3%) were diabetic patients. Diabetic recipients were compared with a control group of 161 nondiabetic recipients matched for age, sex, cause of heart failure, United Network for Organ Sharing status, and immunosuppression era. Outcome measures included posttransplantation survival, incidence of infection, rejection, and transplant coronary artery disease. RESULTS: There was no statistically significant difference in survival between diabetic and nondiabetic recipients, with actuarial survival at 1, 5, and 10 years of 89.3%, 66.9%, and 45.6%, respectively, for diabetic patients and 87.4%, 78.8%, and 59.1%, respectively, for nondiabetic patients (P =.168). There was no significant difference in freedom from infection, rejection, or transplant coronary artery disease between the groups. By using Cox proportional hazard models, development of infection, rejection, and transplant coronary artery disease were independent predictors of decreased survival (P <.001, P =.004, and P =.004, respectively). CONCLUSIONS: These results demonstrate similar short-term and long-term survivals, as well as similar risks for infection and transplant coronary artery disease, in diabetic and nondiabetic patients undergoing cardiac transplantation. The trend toward worse survival in the diabetic cohort, however, raises the possibility that if a greater number of diabetic patients were evaluated, a significant difference in survival might be observed, suggesting the need for a multicenter analysis to validate these outcomes. PMID- 15116013 TI - Twenty-year experience of lung transplantation at a single center: Influence of recipient diagnosis on long-term survival. AB - OBJECTIVES: The objective of this study was to examine the long-term patient outcomes of lung transplantation in a single center. METHODS: Between 1983 and 2003, 521 lung transplants were performed in 501 patients. Major indications were cystic fibrosis (n = 124), chronic obstructive pulmonary disease (n = 88), alpha 1 antitrypsin deficiency (n = 63), pulmonary fibrosis (n = 97), primary pulmonary hypertension (n = 35), Eisenmenger syndrome (n = 21), and miscellaneous end-stage lung diseases (n = 93). RESULTS: The 5-, 10-, and 15-year survivals for all recipients were 55.1% (95% confidence interval: +/-5%), 35.3% (+/-6%), and 26.5% (+/-11%), respectively. The most common causes of death were sepsis and bronchiolitis obliterans syndrome. Despite an increased postoperative mortality rate, patients with primary pulmonary hypertension achieved the best long-term survival (10-year survival: 59%). Recipients with cystic fibrosis without Burkholderia cepacia infection achieved significantly better long-term survival (10-year survival: 52%) than those with Burkholderia cepacia infection (10-year survival: 15%). The 10-year survival was also significantly better in recipients with chronic obstructive pulmonary disease (43%) than in recipients with alpha-1 antitrypsin deficiency (23%). Although the incidence of bronchiolitis obliterans syndrome was similar between recipients with chronic obstructive pulmonary disease (39%) and alpha-1 antitrypsin deficiency (46%), recipients with alpha-1 antitrypsin deficiency died of sepsis more frequently than recipients with chronic obstructive pulmonary disease (27% vs 6%, respectively; P =.0003). CONCLUSIONS: Although bronchiolitis obliterans syndrome and sepsis still limit the durability of the benefit, lung transplantation returns many patients with end-stage lung disease to active and productive lives. Differences in the complications and long-term survival show the important contribution of the recipient diagnosis to the success of lung transplantation. PMID- 15116014 TI - Early tumor necrosis factor-alpha release from the pulmonary macrophage in lung ischemia-reperfusion injury. AB - OBJECTIVE: Tumor necrosis factor-alpha is a proinflammatory mediator required for the development of experimental lung ischemia-reperfusion injury. The alveolar macrophage is a rich source of tumor necrosis factor-alpha in multiple models of acute lung injury. The present study was undertaken to determine whether the alveolar macrophage is an important source of tumor necrosis factor-alpha in lung ischemia-reperfusion injury and whether suppression of its function protects against injury. METHODS: Left lungs of Long-Evans rats underwent normothermic ischemia for 90 minutes and reperfusion for up to 4 hours. Treated animals received gadolinium chloride, a rare earth metal that inhibits macrophage function. Injury was quantitated via lung tissue neutrophil accumulation (myeloperoxidase content), lung vascular permeability, and bronchoalveolar lavage fluid leukocyte, cytokine, and chemokine content. Separate samples were generated for immunohistochemistry. RESULTS: Tumor necrosis factor-alpha secretion occurred at 15 minutes of reperfusion and was localized to the alveolar macrophage by immunohistochemistry. In gadolinium-treated animals, lung vascular permeability was reduced by 66% at 15 minutes (P <.03) of reperfusion and by 34% at 4 hours (P <.02) of reperfusion. Suppression of macrophage function resulted in a 35% reduction in lung myeloperoxidase content (P <.03) and similar reductions in bronchoalveolar lavage leukocyte accumulation. Tumor necrosis factor-alpha and microphage inflammatory protein-1alpha protein levels were markedly reduced in the bronchoalveolar lavage of gadolinium-treated animals by enzyme-linked immunosorbent assay. CONCLUSIONS: The alveolar macrophage secretes tumor necrosis factor-alpha protein by 15 minutes of reperfusion, which orchestrates the early events that eventually result in lung ischemia-reperfusion injury at 4 hours. Gadolinium pretreatment markedly reduces tumor necrosis factor-alpha elaboration, resulting in significant protection against lung ischemia-reperfusion injury. PMID- 15116015 TI - Thyroid dysfunction after pediatric cardiac surgery. PMID- 15116016 TI - Stage III empyema caused by Actinomyces meyeri: a plea for decortication. PMID- 15116017 TI - Thoracic endometriosis: a case report and literature review. PMID- 15116018 TI - Ectopic intra-aortic insertion of a subclavian central venous catheter: case report and review of the literature. PMID- 15116019 TI - Coronary-coronary free internal thoracic artery graft on a single, distal, left anterior descending artery lesion. PMID- 15116020 TI - Thrombolytic therapy for prosthetic valve thrombosis in children: two case reports and review of the literature. PMID- 15116021 TI - Cardiopulmonary bypass elicits a prominent innate immune response in children with congenital heart disease. PMID- 15116022 TI - Modified minimally invasive coronary artery bypass after radical treatment for left breast cancer. PMID- 15116023 TI - Orthotopic mitral valve replacement with autologous pulmonary valve in a porcine model. PMID- 15116024 TI - Direct cerebral perfusion and myocardial protection with moderate systemic hypothermic arrest for high descending aortic aneurysm. PMID- 15116025 TI - Cotton-type collagen insertion at mediastinoscopy for easier node dissection after induction chemoradiotherapy for lung cancer. PMID- 15116026 TI - Safe bilateral use of skeletonized internal thoracic artery in patients with diabetes. PMID- 15116027 TI - Saphenectomy wound complications: the real story. PMID- 15116029 TI - The conversion partial denture: a clinical report. AB - The treatment alternative described maximizes the benefit of remaining teeth while allowing simplified alteration of the prosthesis if abutments are lost during the life span of the removable partial denture (RPD). A conversion partial is an RPD whose tooth-frame assembly components are individually fabricated and then joined with an acrylic resin major connector. The conversion RPD optimizes retention and stabilization of a terminal dentition and can be easily converted to an immediate complete denture. PMID- 15116030 TI - Rehabilitation of an irradiated mandible after mandibular resection using implant/tooth-supported fixed prosthesis: a clinical report. AB - Patients undergoing mandibular resection often have facial asymmetry and cosmetic disfigurement, which include a retruded, deviated mandible, motor and sensory deficiencies, and abnormal intermaxillary relationship and occlusion. An implant supported fixed prosthesis can be an optimal treatment modality. However, there is a problem in creating a repeatable, stable plane of occlusion and maxillomandibular relationship. This report describes the treatment sequence and considerations to rehabilitate a patient undergoing mandibular resection and radiotherapy with an implant-supported fixed prosthesis. PMID- 15116031 TI - Hybrid gate design frameworks for the rehabilitation of the maxillectomy patient. AB - A removable partial denture (RPD) for the restoration of maxillary defects must incorporate sound design principles to reduce potential damage to the abutment teeth and supporting periodontal tissue, while enabling acceptable functional levels of speech, mastication, and deglutition. These functional demands and potential stresses on the abutment teeth may exceed those encountered by patients without maxillary defects. This article describes the indications and design considerations for a hybrid gate design framework that incorporates both conventional cast direct retainers and the gate design concept in the same framework. PMID- 15116032 TI - Influence of occlusal forces on stress distribution in preloaded dental implant screws. AB - STATEMENT OF PROBLEM: Abutment and prosthetic loosening of single and multiple screw-retained, implant-supported fixed partial dentures is a concern. PURPOSE: The purpose of this study was to investigate stress distribution of preloaded dental implant screws in 3 implant-to-abutment joint systems under simulated occlusal forces. MATERIAL AND METHODS: Three abutment-to-implant joint systems were simulated by using the 3-dimensional finite element analysis method: (1) Branemark external hexagonal screw-retained abutment, (2) ITI 8-degree Morse tapered cemented abutment, and (3) ITI 8-degree Morse tapered plus internal octagonal screw-retained abutment. A thermal load and contact analysis method were used to simulate the preload resulting from the manufacturers' recommended torques in implant screw joint assemblies. The simulated preloaded implants were then loaded with 3 simulated static occlusal loads (10 N; horizontal, 35 N; vertical, 70 N; oblique) on the crown position onto the implant complex. RESULTS: Numeric and graphical results demonstrated that the stresses increased in both the abutment and prosthetic screws in the finite element models after simulated horizontal loading. However, when vertical and oblique static loads were applied, stresses decreased in the external hexagonal and internal octagonal plus 8-degree Morse tapered abutment and prosthetic screws with the exception of the prosthetic screw of ITI abutment after 70-N oblique loading. Stresses increased in the ITI 8 degree Morse tapered cemented abutment after both vertical and oblique loads. CONCLUSION: Although an increase or decrease was demonstrated for the maximum calculated stress values in preloaded screws after occlusal loads, these maximum stress values were well below the yield stress of both abutment and prosthetic screws of 2 implant systems tested. The results imply that the 3 implant-to abutment joint systems tested may not fail under the simulated occlusal forces. PMID- 15116033 TI - Effect of lateral cyclic loading on abutment screw loosening of an external hexagon implant system. AB - STATEMENT OF PROBLEM: Efforts to reduce the recurrence of abutment screw loosening with single tooth implant-supported restorations have been reported. However, the current knowledge about the role of the implant external hexagon is incomplete. PURPOSE: This in vitro study investigated the effect of lateral cyclic loading with different load positions on abutment screw loosening of an external hexagon implant system. MATERIAL AND METHODS: Fifteen Branemark implant assemblies were divided equally into 3 groups, A, B, and C. Each assembly consisted of a Mark IV implant (4 x 10 mm) mounted in a brass block, a CeraOne abutment (3 mm), and an experimental cement-retained superstructure. For group A, a cyclic load of 50 N was applied centrally and perpendicular to the long axis of the implant, whereas for group B, the same load was applied eccentrically (at a distance of 4 mm) in a loosening direction. A target of 1.0 x 10(6) cycles (40 months of simulated function) was defined. Group C (control) was left unloaded for the same loading time period as groups A and B. Reverse torque was recorded before and after loading and the difference was calculated. The data were analyzed with 1-way analysis of variance and compared with the Tukey test (alpha=.05). RESULTS: Group A exhibited a significant difference in the reverse torque difference values ([-5.6 to -3.4] +/- 0.86 N.cm) compared with groups B ([ 1.9 to 0.5] +/- 0.99 N.cm) and C ([-0.7 to 0.0] +/- 0.26 N.cm) (P<.001). Groups B and C were not significantly different from each other. CONCLUSION: Within the limitations of this study, reverse torque values of the screw joint were preserved under eccentric lateral loading, as compared with centric loading (P<.001). PMID- 15116034 TI - Diametral tensile strength of a resin composite core with nonmetallic prefabricated posts: an in vitro study. AB - STATEMENT OF PROBLEM: A number of prefabricated nonmetallic posts are currently available for use in conjunction with resin composite cores before fabrication of crowns for endodontically treated teeth. Information is needed regarding the strength of the composite and the nature of attachment between its components. PURPOSE: The aim of this study was to determine the influence of different types of posts on the fracture resistance of a resin composite core material using the diametral tensile strength (DTS) test. MATERIAL AND METHODS: Cylindrical specimens, 6 mm in diameter and 3 mm high, were prepared from resin composite (Tetric Ceram) and a group of prefabricated posts (n=10) as follows: resin composite only (control); Vectrispost (VTS); FiberKor (FKR); AEstheti-Plus post (ATP); Light-Post (LTP); Dentorama post (DRM), and Para-Post (PRP) as a second control. Specimens were stored for 7 days in water at 37 degrees C and then subjected to DTS test in a universal testing machine until failure occurred and load was recorded (N). Mean values and SD for DTS values (MPA) were calculated, and data were analyzed statistically with 1-way analysis of variance, followed by the Tukey test (alpha=.05). Representative specimens from each group were examined with SEM to determine nature of failure. RESULTS: Mean values (SD) in MPa for DTS were as follow: CONTROL GROUP: 49.64 (3.36); VTS: 29.77 (3.36); FKR: 31.9 (2.39); ATP: 28.92 (2.2); LTP: 34.26 (3.37); DRM: 33.45 (2.46), and PRP: 27.90 (2.40). Analysis of variance indicated significant differences among the groups (P<.05). SEM examination indicated that for PRP failure was adhesive in nature, whereas with all nonmetallic posts, cohesive failure was more predominant. CONCLUSION: The use of posts did not result in reinforcement of resin composite core when diametral tensile force was applied. When used with the core material, LTP, DRM, and FKR resulted in the highest DTS values, whereas PRP resulted in the lowest values. PMID- 15116035 TI - Fracture resistance and reliability of new zirconia posts. AB - STATEMENT OF PROBLEM: The radicular portion of zirconia endodontic posts often need to be reshaped to achieve a definitive form and may be airborne-particle abraded to improve adhesion during luting. Therefore, the surface of the tetragonal zirconia ceramics may be transformed and damaged, influencing the mechanical properties of the material. PURPOSE: This study compared the fracture resistance of prefabricated zirconia posts with a new retentive post-head after different surface treatments. MATERIAL AND METHODS: Experimental zirconia posts of 2 different diameters, 1.3 mm and 1.5 mm, were produced from commercially available zirconia powder. A cylindro-conical outline form was used for the root portion of the system and a post-head with 3 retentive rings was designed. Sixty posts of each diameter were divided into 3 groups (n=20). Group 1 was ground with a coarse grit diamond bur; Group 2 was airborne-particle abraded with 110-microm fused alumina particles, and Group 3 was left as-received (controls). Posts were luted into the root-shaped artificial canals with the Clearfil adhesive system and Panavia 21 adhesive resin luting agent. The posts were loaded in a universal testing machine at an inclination of 45 degrees with the constant cross-head speed of 1 mm/min. The fracture load (N) necessary to cause post fracture was recorded, and the statistical significance of differences among groups was analyzed with 1-way ANOVA followed by the Fischer LSD test (alpha=.05). The variability was analyzed using Weibull statistics. RESULTS: Load to fracture values of all zirconia posts depended primarily on post diameter. Mean fracture loads (SD) in Newtons were 518.4 (+/-101.3), 993.6 (+/-224.1), and 622.7 (+/ 110.3) for Groups 1 through 3, respectively, for thicker posts, and 385.9 (+/ 110.3), 627.0 (+/-115.1), and 451.2 (+/-81.4) for Groups 1 through 3, respectively, for thinner posts. Airborne-particle-abraded posts exhibited significantly higher resistance to fracture (P<.05) than those in the other 2 groups for diameters 1.3 mm and 1.5 mm. Grinding reduced Weibull modulus compared with controls, and the values were 4.1 and 6.5 for thicker and thinner posts, respectively. CONCLUSION: Within the limitations of this study, the results suggest that grinding leads to a significant drop in load to fracture of zirconia posts, whereas airborne-particle abrasion increased the fracture load. PMID- 15116036 TI - Shear strength of core-veneer interface in bi-layered ceramics. AB - STATEMENT OF PROBLEM: Delamination of veneering porcelain from underlying ceramic substrates has been reported for all-ceramic restorations. Whether this phenomenon is an inherent weakness of the veneering porcelain due to a weak interface between the veneering and the core porcelains, or merely a fracture through the veneering porcelain itself, has not been explored. PURPOSE: The purpose of this study was to investigate the strength of the substructure and veneering porcelain interface in all-ceramic systems. METHODS: The all-ceramic systems tested with their respective veneering porcelains were IPS-Empress2 with Eris (IE), Procera AllCeram with AllCeram (PA), Procera AllZircon with CZR (PZ), and DC-Zircon with Vita D (DC). The veneering porcelain recommended by the manufacturer for each material was fired to the ceramic core. A metal ceramic (MC) combination was tested as a control group. Sixty specimens, 12 for each system and control, were made from 1 master die. A cylinder of veneering porcelain 2.4 mm in diameter was applied using a specially designed aluminum split mold. After firing, the specimens were placed in a mounting jig and subjected to shear force in a universal testing machine. Load was applied at a crosshead speed of 0.50 mm/min until failure. Average shear strengths (MPa) were analyzed with a 1-way analysis of variance and the Tukey test (alpha=.05). The failed specimens were examined microscopically at original magnification x20 to classify the mode of failure as cohesive in the core, cohesive in the veneer, or adhesive at the interface. RESULTS: The mean shear strengths (+/-SD) in MPa were MC control 30.16 +/- 5.88; IE bonded to Eris 30.86 +/- 6.47; PZ bonded to CZR 28.03 +/- 5.03; DC bonded to Vita D 27.90 +/- 4.79; and PA bonded to AllCeram 22.40 +/- 2.40. IE, PZ, and DC were not significantly different from the MC control. Microscopic examination showed that adhesive failure, or complete delamination, did not occur between the compatible ceramic core and veneering materials. Failure primarily occurred near the interface with residual veneering porcelain remaining on the core. IE with Eris exhibited cohesive failure in both the core and the veneer. CONCLUSION: The bond strengths of 3 of the tested all ceramic materials (IE, PZ, and DC) were not significantly different from the control (MC) group. PMID- 15116037 TI - In vitro evaluation of shear bond strengths of resin to densely-sintered high purity zirconium-oxide ceramic after long-term storage and thermal cycling. AB - STATEMENT OF PROBLEM: The few available studies on the resin bond to zirconium oxide ceramic recommend airborne-particle abrasion and modified resin luting agents containing adhesive monomers for superior and long-term durable bond strengths. It is unknown whether this regimen can also be successfully applied to the intaglio surface of a commercial zirconia-based all-ceramic system. PURPOSE: The purpose of this study was to evaluate and compare bond strengths of different bonding/silane coupling agents and resin luting agents to zirconia ceramic before and after artificial aging. MATERIAL AND METHODS: Composite cylinders (2.9 mm x 3.0 mm) were bonded to airborne-particle-abraded intaglio surfaces of Procera AllZirkon specimens (n=80) with either Panavia F (PAN) or Rely X ARC (REL) resin luting agents after pretreatment with Clearfil SE Bond/ Porcelain Bond Activator (Group SE). In another group, Rely X ARC was used with its bonding/silane coupling agent (Single Bond/Ceramic Primer, Group SB). PAN without any bonding/silane agent (Group NO) was the control. Subgroups of 10 specimens were stored in distilled water for either 3 or 180 days before shear bond strength was tested. One hundred eighty-day-old specimens were repeatedly thermal cycled for 12,000 cycles between 5 and 60 degrees C with a 15-second dwell time. Data were analyzed with 1- and 2-way analysis of variance and the Tukey multiple comparisons test (alpha=.05). Failure modes were examined under original magnification x25. RESULTS: After 3 days, SE-REL (25.15 +/- 3.48 MPa) and SE-PAN (20.14 +/- 2.59 MPa) groups had significantly superior mean shear bond strengths (P=.0007) compared with either NO-PAN (17.36 +/- 3.05 MPa) or SB-REL (16.90 +/- 7.22 MPa). SE-PAN, NO-PAN, and SB-REL groups were not significantly different. Artificial aging significantly reduced bond strengths. After 180-day storage, SE PAN (16.85 +/- 3.72 MPa), and SE-REL (15.45 +/- 3.79 MPa) groups demonstrated significantly higher shear bond strengths than NO-PAN (9.45 +/- 5.06 MPa) or SB REL (1.08 +/- 1.85 MPa) groups. The modes of failure varied among 3-day groups but were 100% adhesive at the ceramic surfaces after artificial aging. CONCLUSION: Artificial aging significantly reduced bond strength. A bonding/silane coupling agent containing an adhesive phosphate monomer can achieve superior long-term shear bond strength to airborne-particle- abraded Procera AllZirkon restorations with either one of the 2 resin luting agents tested. PMID- 15116038 TI - Dentists' molar restoration choices and longevity: a web-based survey. AB - STATEMENT OF PROBLEM: The increasing trend toward esthetics has led to controversy as to the longevity of esthetic restorations and the wisdom of having traditional restorations replaced for esthetic reasons. With the assumption that the way dentists treat their own teeth may be a good indication of the accumulated wisdom of the profession, knowledge of dentists' own restorative choices may provide guidance in selecting the most appropriate restorations. PURPOSE: The purpose of this article was to determine restorative choices of dentists for personal molars and estimate restoration longevity. MATERIAL AND METHODS: Information about dentists' molars was obtained from a Web-based survey designed to receive dentists' demographic data and charting of 8 molar teeth with estimated longevity. Approximately 12,000 e-mails asking to access the website were sent to dentists' addresses randomly selected from a commercial database. Nonrespondents received 1 reminder. The responses were statistically analyzed with a chi-square analysis (alpha=.05). RESULTS: Seven hundred fifty-seven valid replies provided information for 6,034 teeth (22% unrestored, 75% restored, 2% missing). Restorations reported included amalgam (36%), gold inlay/onlay (13%), complete veneer crown (CVC) (10%), metal-ceramic crown (MCC) (8%), and composite (7%). Other esthetic options accounted for less than 3%. Restorations with more than 20 years longevity included amalgam restorations (58%), gold inlays/onlays (48%) and crowns (23%). Fifty-six percent of esthetic restorations and 5% of amalgams were placed in the last 5 years. The following restorations were placed in the last year (n=186): 38% esthetic, 29% crowns (18% MCC; 11% CVC), and 17% gold inlay/onlay or amalgam. Significant differences (P<.001) were identified for dentist's gender, year of graduation and practice location. CONCLUSIONS: Most dentists have not replaced traditional metallic restorations with esthetic alternatives. Dentists still choose nonesthetic options for significant numbers of their own restorations. PMID- 15116039 TI - In vitro effect of microwave irradiation on the retentive force of magnets. AB - STATEMENT OF PROBLEM: Few studies have addressed the possible effect(s) of microwave irradiation on the magnetic properties of "permanent" magnets during the fabrication of dental and maxillofacial prostheses. PURPOSE: The objective of this study was to investigate the influence of microwave irradiation energy on the retentive force of a magnetic attachment system used in maxillofacial prosthetic rehabilitation. MATERIAL AND METHODS: A heat-polymerized PMMA disk (6 cm in diameter) was fabricated. Seven magnets were placed around the wafer in a circumferential fashion: 1 in the center and 6 surrounding it. The 7 magnets were spaced 2 cm from their respective centers. Seven heat-polymerized PMMA cylinders were also used, and a magnet (counter-magnet) was placed in the center of each even with the cylinder's surface. Once the investment had set (45 minutes after mixing), the 7 counter-magnets in the cylinders were placed against the 7 magnets in the acrylic wafer. A second mixture of investment material was added. The flasks were separated, and the acrylic wafer was removed to accommodate the 7 counter-magnets in the base mold in the same geometric configuration and to serve as a "spacer" for the silicone material. A 1:1 mixture of medical grade elastomer (MDX4-4210) and medical adhesive silicone (type A) was packed and compressed, the molds were reclamped, and the excess silicone was removed. The elastomer/silicone wafer was packed and compressed into the test and base molds, the molds were reclamped, and excess silicone was removed. The first group of magnets, designated Group A, received microwave irradiation for 5 minutes at low power (112 W). This procedure was repeated for each group of magnets at the following polymerizing times (n=14): Group B, 10 minutes; Group C, 15 minutes; Group D, 20 minutes; Group E, 25 minutes; Group F, 30 minutes; and Group G, 35 minutes. Measurements of retentive force (N) at 10 mm/min ramp rate of speed of separation was conducted. The specimen rate read 5.0 points/second. Data were analyzed using a 1-way analysis of variance (alpha=.05); individual mean values were compared using the Tukey test (alpha=.05). RESULTS: There were no statistical differences in retentive force between groups D, E, and F (20, 25, and 30 minutes, respectively) or between groups A, B, C, D, F, and G (5, 10, 15, 20, 30, and 35 minutes, respectively). When the microwave-irradiated groups A through G were compared with the control group, there was a significant difference (P<.05) in retentive force (N). Group E (25 minutes) showed the largest reduction of retentive force (0.3 N, a reduction of 12%). CONCLUSION: If a prosthesis is processed using a microwave and contains samarium cobalt magnets, the retentive force may be reduced up to 12% under specific conditions. PMID- 15116040 TI - Comparison of cast Ti-Ni alloy clasp retention with conventional removable partial denture clasps. AB - STATEMENT OF PROBLEM: The elastic property of Ti-Ni alloy may be a desirable property for cast removable partial dentures. However, little is known about the retentive properties of cast Ti-Ni alloy clasps. PURPOSE: This in vitro study investigated the retentive force of various types of clasps during repeated cycles of placement and removal to determine whether Ti-Ni alloy clasps maintain their initial retentive force under varied conditions, including different retentive undercut depths and clasp size (thickness and width). MATERIAL AND METHODS: The test models were developed using a mandibular dentiform with a horizontal plane of occlusion. Two complete metal abutment crowns were made for the left second molar, differing only in retentive undercut depths. A total of 98 clasps (n=7) were fabricated, including 14 wrought wire clasps and 28 clasps from each of the following alloys: Co-Cr alloy (Biosil), Type IV gold alloy (Cast-4), and Ti-Ni alloy (TN-10). Clasps were made to engage one of 2 retentive undercut depths (0.25 mm or 0.75 mm) and were made in 2 sizes (0.8 mm or 1.4 mm). Each clasp was radiographically examined for casting defects and porosity. The force (N) required to remove the clasps was measured using a universal testing machine with a crosshead speed of 10 mm/min. After measuring the retentive force, a masticatory simulator was used to cycle the clasps on and off the metal crown 500 times, simulating the insertion and removal of a removable partial denture clasp. The cycling sequence was repeated 10 times, and retention force was measured after each sequence. The internal surface of the clasp arms was examined for evidence of metal fatigue using scanning electron microscopy. Data were subjected to 2-way and 4-way analysis of variance, followed by Scheffe's multiple comparison test (alpha=.05). RESULTS: Co-Cr alloy and gold alloy clasps in the 0.25-mm retentive undercut groups experienced a gradual decrease in retentive force measurements (P<.001). In contrast, the Ti-Ni alloy clasps maintained a retentive force of approximately 1.8 N and 2.6 N for the 0.8 mm and 1.4 mm clasp groups, respectively. Although Ti-Ni alloy clasps' retentive force was reduced slightly after the first cycling sequence, it was negligible compared with other clasps tested. The wrought wire clasps also retained their retentive force until the final cycling sequence. A similar trend was found in the clasp groups engaging 0.75-mm retentive undercuts. CONCLUSION: Although the end-point retention for all the clasps was similar, there was less change in the retentive force of the cast Ti-Ni alloy clasps after repeated cycling sequences of simulated placement and removal. PMID- 15116041 TI - Fabrication of a closed hollow obturator. AB - This article describes a technique for fabricating a closed hollow bulb obturator. It allows for control of the bulb's wall thickness and weight while not requiring any additional materials or time-consuming steps to the conventional processing procedures. PMID- 15116042 TI - A variation on split-cast mounting for complete denture construction. AB - This article describes a variation of the split-cast mounting technique wherein the border of the definitive cast is wrapped with masking tape to form a container for the dental plaster normally used to affix the cast to the articulator. The entire inferior surface of the cast is coated with a thin film of petroleum jelly, and the cast is mounted in the articulator. After the dental plaster has set, the cast is retained by means of the masking tape. The cast is separated from the dental plaster simply by removing the masking tape. PMID- 15116043 TI - In vivo quantitative evaluation of tooth color with hand-held colorimeter and custom template. AB - This article presents a technique for quantitatively evaluating the color of teeth, as well as color change in restorations and tooth surfaces. Through use of a custom template made of a thermoplastic polymer and a dental colorimeter, tooth surface color can be recorded periodically at the same location intraorally. PMID- 15116044 TI - A procedure for fitting a fixed partial denture to an existing removable partial denture. AB - An indirect technique for fitting a new cast gold crown or fixed partial denture (FPD) to an existing removable partial denture (RPD) is presented. This method uses an acrylic resin coping made on a definitive cast fit directly to the patient's RPD intraorally, with the new FPD subsequently completed on the cast. The patient does not have to relinquish the RPD for laboratory procedures, although an appointment is required to fit the coping intraorally where contact is made with the RPD. PMID- 15116045 TI - A technique for making a diagnostic and surgical template. AB - Implant-supported restorations can restore function and esthetics provided that the implants are placed in the predetermined location and properly angulated. This article describes the fabrication of a diagnostic template incorporating a silicone radiopaque marker as a guide for achieving 3-dimensional evaluation of bone without artifacts using computed tomography. The diagnostic template may be easily transformed into a surgical template by removing the silicone marker. PMID- 15116046 TI - A modified periapical radiographic holder used for standardized implant assessment. PMID- 15116047 TI - Simplifying the use of a reline jig. PMID- 15116048 TI - Cytochrome P450 2D6 and antidepressant toxicity and response: what is the evidence? PMID- 15116049 TI - CYP2C9 and oral anticoagulation therapy with acenocoumarol and warfarin: similarities yet differences. PMID- 15116050 TI - 3-Hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) and genetic variability (single nucleotide polymorphisms) in a hepatic drug uptake transporter: what's it all about? PMID- 15116051 TI - CYP2D6 genotype: impact on adverse effects and nonresponse during treatment with antidepressants-a pilot study. AB - OBJECTIVE: Treatment with antidepressants is frequently associated with adverse effects or insufficient clinical response. Several antidepressants are metabolized by cytochrome P450 (CYP) 2D6. The activity of this enzyme markedly varies among individuals from poor to ultrarapid metabolism on the basis of the polymorphism of the CYP2D6 gene. This association study investigated whether the CYP2D6 genotype distribution differs from that of the German white population either in patients with marked adverse effects or in nonresponders during treatment with antidepressants metabolized by CYP2D6. METHODS: By use of a retrospective, naturalistic approach, outpatient practices and hospitals in southern Germany were asked to report on patients who either had had adverse drug effects or were nonresponsive during treatment with CYP2D6-dependent antidepressants. CYP2D6 genotyping was performed by a panel of polymerase chain reaction techniques. Poor and intermediate metabolizer alleles, as well as allelic duplications of CYP2D6, were detected. RESULTS: Of 28 patients with adverse effects during treatment with a CYP2D6-dependent antidepressant, 8 (29%) had 2 inactive alleles and thus were poor metabolizers. This is a 4-fold increase as compared with the German population (P <.0001). Amplification of fully functional alleles (associated with ultrarapid metabolism) was found in 3 (19%) of the 16 nonresponders (approximately 5.0-fold higher in nonresponders than in the population) (P =.0012). CONCLUSION: The results suggest that the CYP2D6 genotype is associated with the occurrence of adverse effects and clinical nonresponse in psychiatric patients treated with CYP2D6-dependent antidepressants. PMID- 15116052 TI - Acenocoumarol stabilization is delayed in CYP2C93 carriers. AB - OBJECTIVE: Our objective was to assess whether there is an association between the presence of allelic variants of the gene for cytochrome P450 (CYP) 2C9 and anticoagulation problems during the initial 3 to 6 months of acenocoumarol treatment. METHODS: A prospective follow-up study was performed at 2 anticoagulation clinics in the Netherlands. Included subjects started with a standard dose regimen as follows: 6 mg on the first day, 4 mg on the second day, and 2 mg on the third day. CYP2C9 genotypes were assessed, and data on international normalized ratio (INR), comedication, and comorbidity were collected. RESULTS: The CYP2C9 genotype of 231 subjects was assessed. Of these, 147 (63.6%) were wild-type subjects (CYP2C9*1/*1), 38 (16.5%) were carriers of CYP2C9*2, and 46 (19.9%) were carriers of CYP2C9*3. Compared with wild-type subjects, carriers of the CYP2C9*3 allele had (1) a lower chance to achieve stability in the first 6 months of therapy (hazard ratio, 0.62; 95% confidence interval, 0.42-0.91; P <.05) and (2) an increased risk of severe overanticoagulation (INR >6.0) (hazard ratio, 3.80; 95% confidence interval, 1.54 9.39; P <.01). For both outcomes, there was no significant difference between carriers of the CYP2C9*2 allele and wild-type subjects. CONCLUSION: In carriers of the CYP2C9*3 allele more difficulties in terms of stabilization and overanticoagulation were found as compared with wild-type subjects or CYP2C9*2 carriers. CYP2C9 genotyping could be useful to identify potential candidates for more frequent INR controls to minimize problems with acenocoumarol anticoagulation status. PMID- 15116053 TI - Pharmacogenetics of acenocoumarol pharmacodynamics. AB - OBJECTIVE: The aim of this study was to investigate the respective contribution of the different cytochrome P450 (CYP) 2C9 genetic polymorphisms to the interindividual variability of acenocoumarol pharmacodynamic response. METHODS: A total of 263 healthy volunteers were genotyped for CYP2C9*2, CYP2C9*3, CYP2C9*4, and CYP2C9*5 alleles, as well as for the nicotinamide adenine dinucleotide phosphate, reduced, quinone oxidoreductase 1 genetic polymorphism (NQO1*2). Moreover, the 5'-flanking region of the CYP2C9 gene was investigated for new polymorphisms, and haplotype analysis was then performed. Finally, CYP2C9 phenotype was evaluated after a single oral dose of 4 mg of acenocoumarol. Factor VII coagulant activity was measured before and 24 hours after acenocoumarol intake. RESULTS: The CYP2C9*3 allele was the only nonsynonymous single nucleotide polymorphism (SNP) influencing acenocoumarol pharmacodynamics; the percentages of remaining factor VII were 60% +/- 19%, 39% +/- 17%, and 17% for CYP2C9*1/CYP2C9*1, CYP2C9*1/CYP2C9*3, and CYP2C9*3/CYP2C9*3 subjects, respectively (P =.001). Among the white subjects, the CYP2C9 promoter showed the existence of 6 SNPs at positions G-1538A, T-1189C, G-1097A, G-982A, T-640 del, and G-620T with allelic frequencies of 0.085, 0.0398, 0.136, 0.086, 0.005, and 0.0138, respectively. Four major haplotypes could be inferred among white subjects. The haplotype that contains the CYP2C9*3 allele was the only one influencing acenocoumarol pharmacodynamics, explaining 14.3% of its interindividual variability. Body weight explained 5% of acenocoumarol pharmacodynamic variability, whereas the NQO1*2 allele had no significant effect. CONCLUSION: Overall, CYP2C9-related genetic variability accounts for 14% of the interindividual variability in acenocoumarol pharmacodynamic response. The information found by haplotype analysis is mainly related to the CYP2C9*3 SNP. PMID- 15116054 TI - Evidence for inverse effects of OATP-C (SLC21A6) 5 and 1b haplotypes on pravastatin kinetics. AB - OBJECTIVE: We compared the pharmacogenetic effects of OATP-C (organic anion transporting polypeptide C) *1a, *1b (A388G), and *5 (T521C) haplotypes on single dose pharmacokinetics of pravastatin in white subjects. METHODS: Thirty healthy white male subjects were grouped according to their OATP-C haplotype. Each group contained 10 individuals who were either homozygous or heterozygous carriers of the *1a, *1b, or *5 haplotype. After a single oral dose of 40 mg pravastatin, we analyzed kinetic parameters of pravastatin disposition. RESULTS: Values for the area under the plasma concentration-time curve from time 0 to 6 hours [AUC(0-6)] in *1a/*1a, *1a/*1b or *1b/*1b, and *1a/*5 individuals were 114.5 +/- 68.6 microg. L(-1). h, 74.8 +/- 35.6 microg. L(-1). h, and 163.0 +/- 64.6 microg. L( 1). h, respectively, with highly significant differences across all 3 study groups (P =.006) and between subjects carrying the *1b and *5 haplotype (P =.002). Strikingly, values of AUC(0-6) from the OATP-C *1b group were more than 60% lower than those derived from carriers of the wild-type OATP-C *1a haplotype, although this difference failed to reach statistical significance. However, the amount of pravastatin excreted into the urine from time 0 to 12 hours [Ae(0-12)] was significantly diminished in the OATP-C *1b haplotype group (1729 +/- 907 microg) compared with *1a wild-type control subjects (2974 +/- 1590 microg) (P =.049). CONCLUSION: There was a significant effect of tested OATP-C variant haplotypes on pravastatin disposition. Whereas *5 expression delayed the hepatocellular uptake of pravastatin, *1b expression seemed to accelerate OATP-C dependent uptake of the drug. PMID- 15116055 TI - CYP3A5 and MDR1 genetic polymorphisms and cyclosporine pharmacokinetics after renal transplantation. AB - BACKGROUND: The immunosuppressive drug cyclosporine (INN, ciclosporin), whose pharmacokinetic characteristics vary greatly among individuals, is a substrate for cytochrome P450 (CYP) 3A and P-glycoprotein, the product of the multidrug resistance 1 (MDR1) gene. Some of the single nucleotide polymorphisms (SNPs) in these genes are associated with deficient protein expression and reduced in vivo activity. We postulated that, in renal transplant recipients, these SNPs could be associated with interindividual variations in cyclosporine pharmacokinetics. PURPOSE: In 106 renal transplant patients, we evaluated retrospectively the effects of 4 MDR1 SNPs [T-129C, C1236T, G2677(T,A), and C3435T] and of the CYP3A5*1/*3 SNP on cyclosporine pharmacokinetic parameters and exposure indices. RESULTS: The CYP3A5*1 allele was present in 8.5% of patients. The MDR1 C1236T, G2677(T,A), and C3435T SNPs were frequent (17.9%, 18.9%, and 33%, respectively, for the variant homozygous genotype) and exhibited incomplete linkage disequilibrium. None of the cyclosporine pharmacokinetic parameters were associated with the CYP3A5 genetic polymorphism. Patients with the wild-type genotype in MDR1 C1236T SNP had slightly but significantly lower dose-adjusted peak drug concentrations (-16%) (P <.02) and dose-adjusted area under the concentration-time curve (AUC) values over the first 4 hours (-14%) (P <.05) as compared with mutated allele carriers. Haplotype analysis including MDR1 C1236T, G2677(T,A), and C3435T SNPs showed no significant association between haplotypes and cyclosporine pharmacokinetics or systemic exposure, although there was a nonsignificant trend toward higher dose-adjusted AUC values over the first 4 hours and AUC over the 12-hour administration interval for the T-T-T haplotype. CONCLUSION: The presence of the CYP3A5 SNP does not explain the high variability of cyclosporine pharmacokinetics in stable renal transplant patients. Despite the weak association found for the MDR1 C1236T SNP, MDR1 SNPs are unlikely to be useful for cyclosporine dose optimization in clinical practice. PMID- 15116056 TI - Clinical efficacy and toxicity profile of tacrolimus and mycophenolic acid in relation to combined long-term pharmacokinetics in de novo renal allograft recipients. AB - INTRODUCTION: Tacrolimus and mycophenolate mofetil are effective drugs characterized by specific toxicity profiles that may compromise their long-term use in renal transplant recipients. Clinicians, therefore, need reliable drug monitoring tools for relating efficacy and toxicity to drug exposure. Study design We conducted a prospective 12-month pharmacokinetic study of tacrolimus and mycophenolic acid in 100 de novo recipients. The aim was to examine whether tacrolimus and mycophenolic acid exposure parameters (predose trough blood concentration [C(0)], area under the concentration curve from 0 to 12 hours [AUC(0-12)], maximum blood or plasma concentration [C(max)], and dose) would reflect clinical efficacy and toxicity at different time points after transplantation (7 days, 6 weeks, and 3, 6, and 12 months). RESULTS: Initially, after grafting, the tacrolimus AUC(0-12) was higher in recipients with infection (P =.01 on day 7, P =.02 at week 6), whereas the mycophenolic acid AUC(0-12) was not different. There was no difference in tacrolimus exposure between patients who had arterial hypertension or hyperlipidemia and those who did not. Patients with tacrolimus nephrotoxicity received a higher drug dose (P =.03) and had higher drug clearance (P =.02). From 3 months, recipients with anemia or leukopenia had higher mycophenolic acid AUC(0-12) (anemia, P =.03 at month 3 and P =.01 at month 12; leukopenia, P =.01 at month 3 and P =.04 at 1 year) and C(0) (anemia, P =.001 at month 3 and P =.001 at month 12; leukopenia, P =.01 at month 3 and P =.04 at 1 year). Finally, for recipients who did not simultaneously have a target tacrolimus AUC(0-12) of 150 ng x h/mL and a mycophenolic acid AUC(0-12) of 45 mg x h/L by day 7, the incidence of acute rejection tended to be higher (26.3%) compared with patients who reached both target values (7.7%) (P =.07). CONCLUSIONS: Pharmacokinetic exposure parameters of tacrolimus and mycophenolic acid are related to specific drug-induced side effects in a time-dependent fashion. In addition, this study has provided a conceptual basis for defining a combined target therapeutic window for tacrolimus and mycophenolic acid based on sparse AUC(0-12) measurements. PMID- 15116057 TI - Effect of cytochrome P450 3A4 inhibition on the pharmacokinetics of docetaxel. AB - OBJECTIVE: In vitro studies indicate that the anticancer drug docetaxel is primarily eliminated by cytochrome P450 (CYP) 3A4-mediated metabolism. Coadministration of drugs that modulate the activity of CYP3A4 is, therefore, likely to have undesirable clinical consequences. We investigated the effects of the potent CYP3A4 inhibitor ketoconazole on the pharmacokinetics of docetaxel in patients with cancer. METHODS: Seven patients were treated in a randomized crossover design with docetaxel (100 mg/m(2)), followed 3 weeks later by docetaxel (10 mg/m(2)) given in combination with orally administered ketoconazole (200 mg once daily for 3 days), or the reverse sequence. Plasma concentration time data were analyzed by noncompartmental analysis. RESULTS: Ketoconazole coadministration resulted in a 49% decrease in clearance of docetaxel (P =.018). The mean (+/-SD) clearance values were 35.0 +/- 11.8 L/h (95% confidence interval, 24.1-45.9 L/h) for docetaxel alone and 18.2 L/h (95% confidence interval, 9.22-27.1 L/h) in the presence of ketoconazole, respectively. The docetaxel clearance ratio in the presence and absence of ketoconazole was weakly related to the area under the curve of ketoconazole (R(2) = 0.529, P =.064). CONCLUSION: Inhibition of CYP3A4 by ketoconazole in vivo results in docetaxel clearance values that have previously been shown to be associated with a several fold increase in the odds for febrile neutropenia at standard doses. Caution should be taken and substantial dose reductions are required if docetaxel has to be administered together with potent inhibitors of CYP3A4. PMID- 15116058 TI - The effect of gemfibrozil on the pharmacokinetics of rosuvastatin. AB - BACKGROUND: Coadministration of statins and gemfibrozil is associated with an increased risk for myopathy, which may be due in part to a pharmacokinetic interaction. Therefore the effect of gemfibrozil on rosuvastatin pharmacokinetics was assessed in healthy volunteers. Rosuvastatin has been shown to be a substrate for the human hepatic uptake transporter organic anion transporter 2 (OATP2). Inhibition of this transporter could increase plasma concentrations of rosuvastatin. The effect of gemfibrozil on rosuvastatin uptake by cells expressing OATP2 was also examined. METHODS: In a randomized, double-blind, 2 period crossover trial, 20 healthy volunteers were given oral doses of gemfibrozil, 600 mg, or placebo twice daily for 7 days. On the fourth morning of each dosing period, a single oral dose of rosuvastatin, 80 mg, was coadministered. Plasma concentrations of rosuvastatin, N-desmethyl rosuvastatin, and rosuvastatin-lactone were measured. In addition, the effect of gemfibrozil on the uptake of radiolabeled rosuvastatin by OATP2-transfected Xenopus oocytes was studied. RESULTS: Gemfibrozil increased the rosuvastatin area under the plasma concentration-time curve from time 0 to the time of the last quantifiable concentration [AUC(0-t)] 1.88-fold (90% confidence interval, 1.60-2.21) and the maximum observed rosuvastatin plasma concentration (C(max)) 2.21-fold (90% confidence interval, 1.81-2.69) compared with placebo. N-desmethyl rosuvastatin AUC(0-t) and C(max) decreased by 48% and 39%, respectively. Pharmacokinetics of rosuvastatin-lactone was unchanged. The in vitro results indicate that the maximum gemfibrozil inhibition of rosuvastatin OATP2-mediated uptake was 50%; the inhibition constant for the inhibitory process was 4.0 +/- 1.3 micromol/L. CONCLUSIONS: Gemfibrozil increased rosuvastatin plasma concentrations approximately 2-fold, which is similar to the effect of gemfibrozil on pravastatin, simvastatin acid, and lovastatin acid plasma concentrations and substantially less than the effect observed for cerivastatin. Gemfibrozil inhibition of OATP2-mediated rosuvastatin hepatic uptake may contribute to the mechanism of the drug-drug interaction. Care is warranted when gemfibrozil is coadministered with rosuvastatin and other statins. PMID- 15116059 TI - The effect of isotretinoin on the pharmacokinetics and pharmacodynamics of ethinyl estradiol and norethindrone. AB - BACKGROUND: Isotretinoin is a known teratogen, and when it is prescribed to women of childbearing potential, 2 forms of contraception must be used, commonly including hormonal contraception. Although isotretinoin and estradiol are metabolized largely by cytochrome P450 (CYP) 3A4 and glucuronidation, the potential for clinical drug interaction, with subsequent pharmacodynamic impact, has not been evaluated. METHODS: We enrolled 26 healthy women who were to receive isotretinoin for the treatment of severe, recalcitrant nodular acne and who were taking or planning to take oral contraceptives. The pharmacokinetics of ethinyl estradiol and norethindrone (INN, norethisterone) (the components of Ortho Novum 7/7/7; Ortho-McNeil Pharmaceutical, Inc, Raritan, NJ) and pharmacodynamic assessments of oral contraceptive effectiveness (concentrations of serum progesterone, luteinizing hormone, and follicle-stimulating hormone) were determined on days 6 and 20 of 2 separate oral contraceptive cycles, before and during isotretinoin treatment. RESULTS: The addition of isotretinoin to the oral contraceptive regimen resulted in small and inconsistent, although statistically significant (P <.04), decreases in the concentrations of both ethinyl estradiol (9% decrease in area under the plasma concentration-time curve from time 0 to 24 hours after the dose on day 6) and norethindrone (11% decrease in maximum plasma concentration on day 20). Isotretinoin did not cause any statistically significant increases in pharmacodynamic markers, although a majority of women had increases in these measures. Although there was no correlation between isotretinoin (or metabolite) levels and oral contraceptive levels (P >.05), there was a correlation between progesterone level and oral contraceptive levels (P <.05). Variability was large for both pharmacokinetic measures (median coefficients of variation of 44%-69% [for each time point within a study period]) and pharmacodynamic measures (median coefficients of variation of 64%-114%). One woman in each study phase, one before and one during isotretinoin treatment, had a progesterone elevation consistent with possible ovulation. No serious or unexpected adverse events were observed. CONCLUSIONS: The small reduction in ethinyl estradiol and norethindrone levels associated with isotretinoin was not associated with any pharmacodynamic changes in our study. The combination of the teratogenic risk of isotretinoin and the large variability of and correlation between oral contraceptive levels and pharmacodynamic measures, however, strongly reinforces the necessity of additional contraceptive methods during concomitant administration of these drugs. PMID- 15116060 TI - Simvastatin treatment for inflammatory attacks of the hyperimmunoglobulinemia D and periodic fever syndrome. AB - Hyperimmunoglobulinemia D (hyper-IgD) and periodic fever syndrome, a hereditary autoinflammatory syndrome, is characterized by lifelong recurrent episodes of fever and inflammation. No effective treatment is known. It is caused by a defect of mevalonate kinase, an enzyme that follows 3'-hydroxy-3'-methylglutaryl coenzyme A (HMG-CoA) reductase in the isoprenoid pathway. We wanted to test the hypothesis that inhibition of HMG-CoA reductase would ameliorate the inflammatory attacks. Six patients with hyper-IgD syndrome and proven mevalonate kinase deficiency were followed up for 2 treatment periods with either simvastatin, 80 mg/d, or placebo for 24 weeks, separated by a 4-week washout period in a double blind fashion. Simvastatin resulted in a drop in urinary mevalonic acid concentration in all patients and decreased the number of febrile days in 5 of 6 patients. No side effects were observed. These data offer preliminary evidence for the hypothesis that simvastatin may improve inflammatory attacks in the hyper IgD syndrome. This highlights the anti-inflammatory properties of HMG-CoA reductase inhibition. PMID- 15116061 TI - Treatment of urinary incontinence with anticholinergics in patients taking cholinesterase inhibitors for dementia. AB - Clinicians often encounter patients with dementia and urge incontinence who might benefit from both an anticholinergic medication and a cholinesterase inhibitor. At first glance, this combination would seem to violate basic principles of geriatric pharmacology, as the drugs appear to be working at cross-purposes and anticholinergic medications are notorious for worsening cognitive function in susceptible patients. A case is presented and discussed in which this combination was clinically effective and pharmacologically sound. PMID- 15116062 TI - Folic acid and neuroblastoma: too soon to tell. PMID- 15116063 TI - Sildenafil citrate significantly improves nocturnal penile erections in sildenafil non-responding patients with psychogenic erectile dysfunction. AB - Effects of sildenafil citrate on nocturnal penile tumescence and rigidity (NPTR) were evaluated among sildenafil non-responding patients with psychogenic erectile dysfunction. All patients (n=30), equally divided into groups I and II, completed four consecutive nights using the RigiScan Plus device. Sildenafil citrate (50 mg) was given in the third night in group I and in the fourth in group II, whereas a placebo was given in the remaining nights. Additional patients (n=12) receiving only a placebo served as a control group. Results of NPTR recordings revealed neither significant differences between the control and non-sildenafil nights of both test groups, nor between the corresponding values of both groups (P>0.05). On the other hand, when sildenafil citrate nights of groups I and II taken together were compared with placebo nights, a significant increase of total events duration (P<0.001), average rigidity of the tip (P<0.05) and base (P<0.01), and rigidity activity unit (RAU) and tumescence activity unit (TAU) of tip and base (P<0.001) was observed. These results suggest that performance anxiety may be responsible for failure of response during awakening. PMID- 15116064 TI - Incidentally diagnosed Peyronie's disease in men presenting with erectile dysfunction. AB - The aim of this study was to analyze characteristics of patients with Peyronie's disease (PD) diagnosed during a standard evaluation for erectile dysfunction (ED) and compare them with patients presenting with the classical complaints of PD. During a 10-y period, a total of 448 patients were evaluated at our two outpatient clinics, directed by the same author (AK). They were divided into two groups: group I consisted of patients, who presented with only ED and were unaware of their penile deformity, and group II consisted of patients with the classical features of the disease. The clinical characteristics, penile deformities, erectile status and the presence of comorbidities were determined in the two groups. Of 448 Peyronie's patients, 16% (n=71) were detected during diagnostic work-up for ED. In this group of patients, ED was the presenting symptom for a mean period of 31.3+/-9.7 months. The mean age of men was 57.54+/ 8.75 and 52.21+/-10.27 y in groups I and II, respectively (P=0.0001). The mean degree of deformity was 31.5+/-12.66 degrees in group I and 41.16+/-19.14 degrees in group II (P=0.0001). In group I (n=71), 69% (n=49) of the patients had a poor erectile response to the combined injection and stimulation (CIS) test. Also, in this group, the mean degrees of deformity in CIS-positive and -negative patients were 27.05+/-12.50 and 33.80+/-12.03 degrees , respectively (P=0.033). Diabetes mellitus (40%) was the leading comorbidity in group I, while at least one comorbidity was observed in 73% of the cases (P=0.001). A remarkable percent of Peyronie's patients (16%) were detected during a standard evaluation for ED. This study analyzed, for the first time, the frequency and the characteristics of incidentally diagnosed Peyronie's patients who presented with only ED. Our data indicate that one should always consider the possibility of PD in older patients with diabetes, presenting with only ED. PMID- 15116065 TI - Regulated, electroporation-mediated delivery of pro-opiomelanocortin gene suppresses chronic constriction injury-induced neuropathic pain in rats. AB - We previously reported that intrathecal pro-opiomelanocortin gene electroporation could reduce pain sensitivity induced by chronic constriction injury (CCI) of the sciatic nerve. For optimal use of antinociceptive gene therapy, it might be important to control the expression of the transfected gene extrinsically. For this purpose, a doxycycline-controlled transrepressor system composed of two plasmids coding, respectively, for pro-opiomelanocortin gene (pTRE2-POMC) and the silencer (pTel-off) was employed. The regulation of beta-endorphin expression was first assessed in spinal neuronal culture, then we electrotranfected this plasmid into the spinal cord of mononeuropathic rats and evaluated the analgesic potential of this therapy in vivo by thermal and mechanical withdrawal latency. Intraperitoneal injections of various doses of doxycycline were made to elucidate the possible exogenous downregulation of transfected beta-endorphin gene expression in vivo. The levels of beta-endorphin were analyzed by intrathecal microdialysis and radioimmunoassay. Intrathecal pTRE2-POMC/pTel-off electroporation elevated spinal beta-endorphin levels, as manifested in a significantly elevated pain threshold for chronic constriction injury limbs. Intraperitoneal doxycycline decreased the antinociceptive effect and spinal beta endorphin levels in a dose-dependent manner. We concluded that intrathecal pTRE2 POMC/pTel-off electroporation alleviates CCI-induced limb pain, and can be controlled by intraperitoneal doxycycline administration. PMID- 15116066 TI - Mosaics of gene variations in the Interleukin-10 gene promoter affect interleukin 10 production depending on the stimulation used. AB - Interleukin-10 (IL-10), a cytokine involved in many aspects of the immune response shows interindividual variations in their expression. However, genetic variations of the 5'-flanking region of the IL-10 gene (PIL-10) are poorly characterised with respect to different stimuli. New extended haplo- and genotypes are identified present at differing frequencies in three geographically separated populations. Their influence on IL-10 expression have been assessed in vitro after stimulation of leukocytes with lipopolysaccharide (LPS), dibutyryl cAMP or following immortalisation with Epstein-Barr virus (lymphoblastoid cell line (LCL)). Interindividual differences of IL-10 production were found to be related to single-nucleotide polymorphisms (SNP) haplotype -6752/-6208 in LCLs (P<0.02), and for haplotypes comprising SNPs -6752/-6208/-3538 after LPS stimulation (P<0.03). Carriers of the IL10.G microsatellite with 22, 24 or 26 dinucleotide repeats linked with the -1087G SNP, exhibited the highest levels of IL-10 expression. Contrasting IL-10 secretion patterns were found for IL10.R microsatellite alleles characterised by 15 dinucleotide repeats: after LPS stimulation this allele was associated with high IL-10 production (P<0.007), but with low IL-10 levels in LCLs (P< 0.038). Thus, the effects of mosaics of genetic elements in the PIL-10 on the capacity of leukocytes to produce IL-10 depend on the agent inducing IL-10 expression. PMID- 15116067 TI - Entry of newly synthesized GLUT4 into the insulin-responsive storage compartment is GGA dependent. AB - Following biosynthesis, both GLUT1 and VSV-G proteins appear rapidly (2-3 h) at the plasma membrane, whereas GLUT4 is retained in intracellular membrane compartments and does not display any significant insulin responsiveness until 6 9 h. Surprisingly, the acquisition of insulin responsiveness did not require plasma membrane endocytosis, as expression of a dominant-interfering dynamin mutant (Dyn/K44A) had no effect on the insulin-stimulated GLUT4 translocation. Furthermore, expression of endocytosis-defective GLUT4 mutants or continuous surface labeling with an exofacial specific antibody demonstrated that GLUT4 did not transit the cell surface prior to the acquisition of insulin responsiveness. The expression of a dominant-interfering GGA mutant (VHS-GAT) had no effect on the trafficking of newly synthesized GLUT1 or VSV-G protein to the plasma membrane, but completely blocked the insulin-stimulated translocation of newly synthesized GLUT4. Furthermore, in vitro budding of GLUT4 vesicles but not GLUT1 or the transferrin receptor was inhibited by VHS-GAT. Together, these data demonstrate that following biosynthesis, GLUT4 directly sorts and traffics to the insulin-responsive storage compartment through a specific GGA-sensitive process. PMID- 15116068 TI - The in vivo role of PtdIns(3,4,5)P3 binding to PDK1 PH domain defined by knockin mutation. AB - We generated homozygous knockin ES cells expressing a form of 3-phosphoinositide dependent protein kinase-1 (PDK1) with a mutation in its pleckstrin homology (PH) domain that abolishes phosphatidylinositol 3,4,5-tris-phosphate (PtdIns(3,4,5)P3) binding, without affecting catalytic activity. In the knockin cells, protein kinase B (PKB) was not activated by IGF1, whereas ribosomal S6 kinase (RSK) was activated normally, indicating that PtdIns(3,4,5)P3 binding to PDK1 is required for PKB but not RSK activation. Interestingly, amino acids and Rheb, but not IGF1, activated S6K in the knockin cells, supporting the idea that PtdIns(3,4,5)P3 stimulates S6K through PKB-mediated activation of Rheb. Employing PDK1 knockin cells in which either the PtdIns(3,4,5)P3 binding or substrate docking 'PIF pocket' was disrupted, we established the roles that these domains play in regulating phosphorylation and stabilisation of protein kinase C isoforms. Moreover, mouse PDK1 knockin embryos in which either the PH domain or PIF pocket was disrupted died displaying differing phenotypes between E10.5 and E11.5. Although PDK1 plays roles in regulating cell size, cells derived from PH domain or PIF pocket knockin embryos were of normal size. These experiments establish the roles of the PDK1 regulatory domains and illustrate the power of knockin technology to probe the physiological function of protein-lipid and protein-protein interactions. PMID- 15116069 TI - Ring-shaped architecture of RecR: implications for its role in homologous recombinational DNA repair. AB - RecR, together with RecF and RecO, facilitates RecA loading in the RecF pathway of homologous recombinational DNA repair in procaryotes. The human Rad52 protein is a functional counterpart of RecFOR. We present here the crystal structure of RecR from Deinococcus radiodurans (DR RecR). A monomer of DR RecR has a two domain structure: the N-terminal domain with a helix-hairpin-helix (HhH) motif and the C-terminal domain with a Cys4 zinc-finger motif, a Toprim domain and a Walker B motif. Four such monomers form a ring-shaped tetramer of 222 symmetry with a central hole of 30-35 angstroms diameter. In the crystal, two tetramers are concatenated, implying that the RecR tetramer is capable of opening and closing. We also show that DR RecR binds to both dsDNA and ssDNA, and that its HhH motif is essential for DNA binding. PMID- 15116070 TI - The Caenorhabditis elegans MAPK phosphatase VHP-1 mediates a novel JNK-like signaling pathway in stress response. AB - Mitogen-activated protein kinases (MAPKs) are integral to the mechanisms by which cells respond to physiological stimuli and to a wide variety of environmental stresses. MAPK cascades can be inactivated at the MAPK activation step by members of the MAPK phosphatase (MKP) family. However, the components that act in MKP regulated pathways have not been well characterized in the context of whole organisms. Here we characterize the Caenorhabditis elegans vhp-1 gene, encoding an MKP that acts preferentially on the c-Jun N-terminal kinase (JNK) and p38 MAPKs. We found that animals defective in vhp-1 are arrested during larval development. This vhp-1 defect is suppressed by loss-of-function mutations in the kgb-1, mek-1, and mlk-1 genes encoding a JNK-like MAPK, an MKK7-type MAPKK, and an MLK-type MAPKKK, respectively. The genetic and biochemical data presented here demonstrate a critical role for VHP-1 in the KGB-1 pathway. Loss-of-function mutations in each component in the KGB-1 pathway result in hypersensitivity to heavy metals. These results suggest that VHP-1 plays a pivotal role in the integration and fine-tuning of the stress response regulated by the KGB-1 MAPK pathway. PMID- 15116071 TI - Ssn6-Tup1 requires the ISW2 complex to position nucleosomes in Saccharomyces cerevisiae. AB - The Imitation SWItch (ISWI) chromatin remodeling factors have been implicated in nucleosome positioning. In vitro, they can mobilize nucleosomes bi-directionally, making it difficult to envision how they can establish precise translational positioning of nucleosomes in vivo. It has been proposed that they require other cellular factors to do so, but none has been identified thus far. Here, we demonstrate that both ISW2 and TUP1 are required to position nucleosomes across the entire coding sequence of the DNA damage-inducible gene RNR3. The chromatin structure downstream of the URS is indistinguishable in Deltaisw2 and Deltatup1 mutants, and the crosslinking of Tup1 and Isw2 to RNR3 is independent of each other, indicating that both complexes are required to maintain repressive chromatin structure. Furthermore, Tup1 repressed RNR3 and blocked preinitiation complex formation in the Deltaisw2 mutant, even though nucleosome positioning was completely disrupted over the promoter and ORF. Our study has revealed a novel collaboration between two nucleosome-positioning activities in vivo, and suggests that disruption of nucleosome positioning is insufficient to cause a high level of transcription. PMID- 15116072 TI - A moderately high intake compared to a low intake of zinc depresses magnesium balance and alters indices of bone turnover in postmenopausal women. AB - OBJECTIVES: To determine whether moderately high or low intakes of zinc adversely affect the copper status of postmenopausal women to result in unfavorable changes in calcium and magnesium metabolism and other indicators of bone turnover. DESIGN: After a 10-day equilibration period in which the diet provided 31.5 micromol (2 mg) Cu and 137.7 micromol (9 mg) Zn/8.4 MJ (2000 kcal), the subjects were randomly divided into two groups, with one group fed the basal diet supplemented to provide 15.7 micromol (1 mg) Cu/8.4 MJ, and the other group fed the same diet supplemented to provide 47.2 micromol (3 mg) Cu/8.4 MJ. After equilibration, both groups were fed the basal diet with no zinc supplemented (provided 45.9 micromol [3 mg] Zn/8.4 MJ) for 90 days; this was followed by another 10-day equilibration period before the basal diet was supplemented with zinc to provide 811 micromol (53 mg)/8.4 MJ for 90 days. SETTING: The metabolic unit of the Grand Forks Human Nutrition Research Center, Grand Forks, ND, USA. SUBJECTS: A total of 28 postmenopausal women recruited by advertisement throughout the United States of America. Among them, 25 women (64.9+6.7 y) completed the study; 21 as designed. RESULTS: The moderately high intake compared to the low intake of zinc increased the excretion of magnesium in the feces and urine, which resulted in a decreased magnesium balance. In the women fed low dietary copper, plasma osteocalcin was higher during the low-zinc than high-zinc dietary period. The urinary excretion of N-telopeptides was increased and the serum calcitonin concentration was decreased by high dietary zinc regardless of dietary copper. CONCLUSIONS: A moderately high intake of zinc (811 micromol/day; 53 mg/day) did not induce changes in copper metabolism that resulted in unfavorable changes in bone or mineral metabolism. However, low dietary zinc (45.9 micromol/day; 3 mg/day) apparently resulted in undesirable changes in circulating calcitonin and osteocalcin. As a moderately high intake of zinc decreased magnesium balance, further study of the possibility that a high intake of zinc is a health concern for individuals consuming less than the recommended amounts of magnesium is warranted. PMID- 15116073 TI - Milk drinking, ischaemic heart disease and ischaemic stroke I. Evidence from the Caerphilly cohort. AB - OBJECTIVE: To test the hypothesis that milk drinking increases the risk of ischaemic heart disease (IHD) and ischaemic stroke in a prospective study. DESIGN: In the Caerphilly Cohort Study dietary data, including milk consumption, were collected by a semiquantitative food frequency questionnaire in 1979-1983. The cohort has been followed for 20-24 y and incident IHD and stroke events identified. SUBJECTS: A representative population sample in South Wales, of 2512 men, aged 45-59 y at recruitment. MAIN OUTCOME MEASURES: In total, 493 men had an IHD event and 185 an ischaemic stroke during follow-up. RESULTS: After adjustment, the hazard ratio in men with a milk consumption of one pint (0.57 l) or more per day, relative to men who stated that they consumed no milk, is 0.71 (0.40-1.26) for IHD and 0.66 (0.24-1.81) for ischaemic stroke. At baseline, 606 men had had clinical or ECG evidence of vascular disease, and in these the vascular risk was even lower (0.37; 0.15-0.90). The hazard ratio for IHD and ischaemic stroke combined is 0.64 (0.39-1.06) in all men and 0.37 (0.15-0.90) in those who had had a prior vascular event. CONCLUSION: The data provide no convincing evidence that milk consumption is associated with an increase in vascular disease risk. Evidence from an overview of all published cohort studies on this topic should be informative. SPONSORSHIP: : The Medical Research Council, the University of Wales College of Medicine and Bristol University. Current support is from the Food Standards Agency. PMID- 15116075 TI - Conclusions about differences in linear growth between Bangladeshi boys and girls depend on the growth reference used. AB - OBJECTIVE: To examine sex differences in height-for-age z-scores and the percentage stunting among Bangladeshi children estimated using three growth references. DESIGN, SETTING AND SUBJECTS: Data collected between 1990 and 1999 by Helen Keller International's nutritional surveillance system in rural Bangladesh were analyzed for 504 358 children aged 6-59 months. Height-for-age z-scores were estimated using the 1977 NCHS, 2000 CDC and 1990 British growth references. RESULTS: The shape of the growth curves for Bangladeshi boys and girls, and their positions relative to one another, depend on which of the three growth references is used. At 6 months of age the British reference showed no sex difference whereas the NCHS and CDC showed girls to have higher average z-scores than boys by 0.14 and 0.28 s.d., respectively. While all references showed a faster deterioration of girls' z-scores from 6 to 24 months, the magnitude and direction of the sex differences, and how they changed with age, were different. There was greater disagreement about girls' z-scores than boys. Discontinuities at 24 months in the NCHS and CDC produced jagged curves whereas the British curves were smooth. CONCLUSIONS: The assessment of sex differences in linear growth depends on the growth reference used. Reasons for the different results need to be determined and may aid the final development of the new WHO international growth reference and the guidelines for its use. The findings suggest that anthropometry as a tool to explore the effects of societal gender inequality must be used with caution. PMID- 15116074 TI - Milk drinking, ischaemic heart disease and ischaemic stroke II. Evidence from cohort studies. AB - OBJECTIVE: Milk consumption is considered a risk factor for vascular disease on the basis of relevant biological mechanisms and data from ecological studies. The aim was to identify published prospective studies of milk drinking and vascular disease, and conduct an overview. DESIGN: The literature was searched for cohort studies, in which an estimate of the consumption of milk, or the intake of calcium from dairy sources, has been related to incident vascular disease. MAIN OUTCOME MEASURES: Ischaemic heart disease and ischaemic stroke. RESULTS: In total, 10 studies were identified. Their results show a high degree of consistency in the reported risk for heart disease and stroke, all but one study suggesting a relative risk of less than one in subjects with the highest intakes of milk. A pooled estimate of relative odds in these subjects, relative to the risk in subjects with the lowest consumption, is 0.87 (95% CI 0.74-1.03) for ischaemic heart disease and 0.83 (0.77-0.90) for ischaemic stroke. The odds ratio for any vascular event is 0.84 (0.78-0.90). CONCLUSIONS: Cohort studies provide no convincing evidence that milk is harmful. While there still could be residual confounding from unidentified factors, the studies, taken together, suggest that milk drinking may be associated with a small but worthwhile reduction in heart disease and stroke risk. SPONSORSHIP: The University of Wales College of Medicine and Bristol University. Current support is from the Food Standards Agency. PMID- 15116076 TI - Evidence for a protective (synergistic?) effect of B-vitamins and omega-3 fatty acids on cardiovascular diseases. AB - The results of dietary intervention trials favor the hypothesis that higher intakes of B-vitamins (folate, vitamin B(6) and B(12)), and subsequently lower total homocysteine (tHcy) concentrations, are causally associated with a decreased risk of vascular disease in patients with cardiovascular diseases (CVD). The same is true for a higher intake of omega-3 fish fatty acids. Yet, the lack of hard end points and/or appropriate study designs precludes a definitive conclusion about causality. In the future, intervention trials with hard end points and randomized double-blind placebo-controlled designs should be able to elucidate the causality problem. There are several pathways by which B-vitamins and omega-3 fatty acids may exert their protective effect on CVD, a common pathway is a beneficial effect on the endothelial function and hemostasis. With respect to synergy between B-vitamins and omega-3 fatty acids, there is no evidence that fish oils have a tHcy-lowering effect beyond the effect of the B vitamins. Nevertheless, animal studies clearly illustrate that vitamin B(6)- as well as folate-metabolism are linked with those of long-chain omega-3 fatty acids. Furthermore, a human study indicated synergistic effects of folic acid (synthetic form of folate) and vitamin B(6) together with omega-3 fatty acids on the atherogenic index and the fibrinogen concentration. Although these results are promising, they were produced in very small selective study populations. Thus, confirmation in large well-designed intervention trials is warranted. PMID- 15116077 TI - Number of days needed to assess energy and nutrient intake in infants and young children between 6 months and 2 years of age. AB - OBJECTIVE: To estimate the minimum number of days of recorded dietary intake needed to place infants and young children into thirds of a population distribution with an acceptable degree of accuracy. DESIGN: Dietary intake data collected from 5-day weighed food records for 72 infants and young children up to 2 y of age, collected during a cross-sectional study, were analysed to estimate the number of recording days necessary to assess intake of energy and 10 nutrients. SETTING: Community study among healthy infants and children. SUBJECTS: Parents attended recruitment sessions in local community areas. In total, 72 subjects were entered into the study and all completed the 5-day dietary assessment period. INTERVENTIONS: A 5-day weighed record of children's dietary intake was made by the parents or regular carer. RESULTS: Mean within subject standard deviations were smaller than mean between subject standard deviations with respect to energy (778 vs 824 kJ/day), macronutrient subclasses: protein (8 vs 9.4 g/day); fat (9 vs 10 g/day); and carbohydrate (26 vs 29 g/day) and for specific micronutrients: calcium (163 vs 236 mg/day); phosphorus (143 vs 270 mg/day); magnesium (25 vs 43 mg/day); iron (2 vs 3 mg/day); zinc (1 vs 1.3 mg/day); ascorbic acid (27 vs 64 mg/day) and retinol equivalents (281 vs 424 microg/day). Estimated number of days of food records necessary to assess intake of energy, protein, fat and carbohydrate with acceptable degree of accuracy were 5,4,4 and 3, respectively. For all the micronutrients included in this analysis 2 days of recording were necessary. CONCLUSION: Compared with adults and older children, fewer days are needed to classify this age group into thirds of the distribution with an acceptable degree of accuracy according to intake of energy and specific nutrients. SPONSORSHIP: The data collection phase of this study was supported by a grant from the Procter and Gamble Company, Cincinnati, OH, USA. PMID- 15116078 TI - Risk factors for coronary heart disease in two similar Indian population groups, one residing in India, and the other in Sydney, Australia. AB - OBJECTIVE: To identify the prevalence of coronary risk factors among South Asian Indians in Australia and India. DESIGN: Cross-sectional intercountry comparison. SUBJECTS: Healthy volunteers aged 23-75 y recruited from the Indian community in Sydney Australia (n=125), and their nominated relatives in India, (n=125). RESULTS: The two groups were of similar background with over 90% of the group in India being siblings, parents or relatives of the group in Australia. There was no difference in the populations between India and Australia with regard to mean age (40+/-11.5 vs 39+/-10.3 y), body mass index (BMI) (25+/-3.3 vs 25+/-3.5 kg/m(2)), lipoprotein (a) (178 vs 202 mg/l), total cholesterol (5.3+/-1.3 vs 5.3+/-1.2 mmol/l) or triglyceride (1.7+/-0.8 vs 1.7+/-0.8 mmol/l). The group in India had higher insulin (median values) (139 vs 83 pmol/l, P=0.0001), waist-to hip ratio (WHR) (0.88+/-0.08 vs 0.85+/-0.09, P=0.01), exercise time (23.7+/-32.7 vs 17.2+/-23.2 h/week, P=0.07), lower waist (83+/-10.0 vs 85+/-11.1 cm, P=0.05) and high-density lipoprotein (0.9+/-0.3 vs 1.1+/-0.6 mmol/l, P=0.02). Women in India had lower BMI (22.7+/-2.9 vs 25.3+/-4.2 kg/m(2), P<0.001), higher insulin (182 vs 90 pmol/l, P<0.001), WHR (0.86+/-0.08 vs 0.77+/-0.06, P<0.001)) and prevalence of abdominal obesity (% WHR >0.8, 73 vs 23%, P<0.001; odds of waist >90 cm=2.3, P<0.05). Men in India had the same BMI, lower waist (85.5+/-8.8 vs 92.9+/-7.2 cm, P<0.001) and WHR (0.89+/-0.09 vs 0.93+/-0.05, P<0.01) but higher insulin (137 vs 76 pmol/l). CONCLUSION: The group in Australia (especially women) have a more favourable disease risk profile than those in India. The fact that the groups are of such similar background and partly related, make it unlikely that changes due to migration have a strong genetic bias. In contrast to other studies, the absence here of excessive weight gain on migration may be a key factor in disease risk prevention. PMID- 15116079 TI - No relations between breast cancer risk and fatty acids of erythrocyte membranes in postmenopausal women of the Malmo Diet Cancer cohort (Sweden). AB - OBJECTIVE: To examine the fatty acid composition of erythrocyte membranes, in relation to obesity indexes and breast cancer risk. DESIGN: A nested case-control study. SETTING: The Malmo Diet Cancer cohort, Sweden. SUBJECTS: Among women 50 y or older at baseline (n=12 803), incident breast cancer cases (n=237) were matched to controls (n=673) on age and screening date. METHODS: A diet history method, a structured questionnaire, anthropometrics and blood samples provided data. Analysis included partial correlation coefficients between dietary fatty acids (DFA) and fatty acids of erythrocyte membranes (EFA), and Spearman's rank order correlations between EFA and four obesity indexes. Conditional logistic regression examined breast cancer risks related to EFA. RESULTS: DFA and EFA from fish and milk, and DFA and EFA linoleic acid, show significant positive associations. Relations are negative between indexes of obesity and "milk" EFA, but positive between indexes of obesity and indexes of delta9- and delta6 desaturase enzyme activity. No significant relations were observed between EFA and breast cancer risk. CONCLUSIONS: Similar to other studies, dietary fish and milk fatty acids, and linoleic acid, are related to the corresponding EFA. Breast cancer risk was not significantly related to EFA in this study. However, the findings suggest positive relations between body mass index, body fat per cent and indexes of desaturase activity, and negative relations between central obesity and milk EFA. SPONSORSHIP: The Swedish Cancer Society, the Swedish Medical Research Council, the European Commission, the Swedish Dairy Association and the City of Malmo. PMID- 15116080 TI - Validity and reproducibility of self-reported intake of fruit and vegetable among 6th graders. AB - OBJECTIVE: Reproducible and valid methods for measuring fruit and vegetable consumption among young school children are urgently needed. The aim of this study was to test the reproducibility and the validity of a newly developed questionnaire on the intake of fruit and vegetable among Norwegian 6th graders. The questionnaire consisted of a 24-h recall part and a food frequency part. DESIGN: A total of 114 pupils completed the questionnaire two times 14 days apart, and another 85 pupils completed the questionnaire and 7-day food diaries. SUBJECTS: Pupils of 6th grade with a mean age of 11.9 y. RESULTS: Spearman correlation coefficients between the frequency part of the questionnaire administered two times varied from 0.62 for fruit to 0.83 for potato, and no difference was seen between the average intakes from the two 24-h recalls. The 24 h recall part of the questionnaire gave higher estimates for the average intake of fruit and juice compared to the 7-day record, while no difference was observed for vegetable intake. Spearman correlation coefficients between the frequency part and the records varied from 0.21 for fruit and potato to 0.32 for the total intake of fruit and vegetable. CONCLUSION: Both the 24-h recall and the frequency part gave a consistent response on separate occasions over the test-retest study period. The 6th graders were capable of recording yesterday's intake of vegetable, but overestimated the intake of fruit and juice. The ability to rank subjects based on the frequency part was rather low. PMID- 15116081 TI - A transferable programme of nutritional counselling for rehabilitation following myocardial infarction: a randomised controlled study. AB - OBJECTIVE: To evaluate the response to simple innovative dietary counselling in post myocardial infarction patients. DESIGN: Randomised controlled trial. SETTING: Cardiac rehabilitation programmes of two acute hospitals in Lanarkshire, Scotland. PATIENTS: A total of 69 men and 29 women aged 35-75 y who survived acute myocardial infarction and participated in the cardiac rehabilitation programmes of the study hospitals between 1st September 1997 and 1st August 1998. RESULTS: At 12 weeks follow-up, diet composition had improved significantly in intervention subjects, but no such change was evident in the control group. The target of five portions of fruit and vegetables per day was achieved by 65% of intervention subjects but only 31% of control subjects (P=0.004). Between-group differences in food intakes were no longer evident at 1 y. Diet composition did however remain in line with current dietary targets in intervention subjects. CONCLUSION: Compared to conventional cardiac rehabilitation, this intervention, focused on targets known to improve mortality, improved diet in post myocardial infarction patients. However, a more sustained programme is required to maintain improvements. Delivery of the intervention was expensive and further research is required to determine the feasibility of a group approach using the same package. SPONSORSHIP: The study was supported by Chief Scientist Office of the Scottish Executive Department of Health. PMID- 15116082 TI - Nutritional intakes of 1072 French free-living men with and without diagnosed cardiovascular risk factors. AB - OBJECTIVE: To compare the dietary intakes of free-living people with and without previously diagnosed diet-modifiable cardiovascular risk factors (hypertension, hypercholesterolaemia and diabetes). DESIGN: Cross-sectional survey on cardiovascular risk factors including a three-consecutive-day food record. SETTING: Multicentre setting in Lille (northern France), Strasbourg (north-east) and Toulouse (south-west) areas. SUBJECTS: A total of 1072 middle-aged men randomly selected from the general population: group 1 (504 men without previously diagnosed diet-modifiable risk factor), group 2 (377 men with one previously diagnosed diet-modifiable risk factor) and group 3 (191 men with two or three previously diagnosed diet-modifiable risk factors). INTERVENTIONS: None. RESULTS: Total daily energy intake equalled 10731 kJ/day (standard error: 119), 9991 (138) and 9737 (166) in groups 1, 2 and 3, respectively (P<0.0001 for ANOVA comparing the three groups), and daily energy intake without alcohol equalled 9860 (115), 9096 (132) and 8654 (159) kJ/day (P<0.0001). The proportion of calories from animal proteins (in daily energy intake without alcohol) increased from group 1 to 3 (P<0.0001), whereas the proportion from oligosaccharides decreased (P<0.0001). The proportion of calories from alcohol (in total daily energy intake) increased with the number of risk factors (P<0.0001). These results remained significant after adjustment for confounders. No significant group differences were found in the proportions of energy from polysaccharides, saturated, monounsaturated and polyunsaturated fats. CONCLUSIONS: As compared with subjects without risk factor, significant quantitative and qualitative changes are observed in individuals with diagnosed hypertension, hypercholesterolaemia or diabetes. However, lower consumptions of saturated fats and alcohol are needed. PMID- 15116083 TI - Subclinical vitamin A deficiency in Israeli-Bedouin toddlers. AB - OBJECTIVE: This study was designed to estimate the prevalence of and evaluate risk factors for subclinical vitamin A deficiency in Arab-Bedouin children at age 18 months, followed from birth. DESIGN: Community-based, prospective, cohort study conducted in Rahat, a large Arab-Bedouin township, located near the city of Beer Sheva in the Negev region of southern Israel. SUBJECTS: Healthy Bedouin infants (n=117) from the township, born at Soroka University Medical Center (SUMC) in Beer Sheva, were randomly recruited at birth. Enrollment was restricted to well infants born weighing >2500 g at birth. RESULTS: More than 15% of the children had serum retinol concentrations below 0.7 micromol/l. Male sex (odds ratio (OR) 4.17 [1.14-15.32], P=0.031), stunting at age 12 months (OR 10.09 [2.00 50.97], P=0.05) and warm season at age 18 months (OR 6.20 [1.36-28.28], P=0.018) were associated with vitamin A deficiency. Maternal education decreased the risk of vitamin A deficiency (OR 0.81 [0.68-0.95], P=0.011). CONCLUSIONS: Study results indicate a significant vitamin A deficiency problem among Bedouin children. Deficiency may be prevented by increasing dietary intake of vitamin A, especially during the warm season. Other interventions include preventing and controlling diarrheal diseases in order to avert nutritional stunting, and providing nutritional education to women of childbearing age. SPONSORSHIP: This study received financial support from the National Institute of Allergy and Infectious Diseases (AI-26497), the US-Israel Bi-national Science Foundation (BSF 90-00257), and the National Academy of Sciences/Institute of Medicine (AID/ANE 0158-G-SS-9035-00). PMID- 15116084 TI - Lycopene and beta-carotene are bioavailable from lycopene 'red' carrots in humans. AB - OBJECTIVE: The purpose of this study was to determine if lycopene and beta carotene are bioavailable from lycopene red carrots and if lycopene absorption is affected by carrot fiber. DESIGN: Two crossover studies in humans attempted to compare the relative bioavailability of lycopene and beta-carotene from tomato paste to a genetically selected lycopene red carrot during chronic feeding. Each study contained three treatment groups. The vehicle of administration was muffins. INTERVENTION AND METHODS: Study 1 (n=9) used white carrots (0 mg lycopene/day), red carrots (5 mg/day), and tomato paste (20 mg/day). Study 2 (n=10) used red carrots (2.6 mg/day), tomato paste (5 mg/day), and tomato paste plus white carrots (5 mg/day). Each intervention lasted 11 days with a 10-day washout period between treatments. Serum lycopene and beta-carotene were measured by HPLC. RESULTS: Statistical analysis indicated a significant effect of muffin type in study 1 (P<0.001), and a significant treatment by sequence interaction in study 2 (P=0.04). The response to increasing amounts of lycopene is linear at the levels fed in these studies (r=0.94). The data suggest that maintenance of serum lycopene concentrations at 0.3 micromol/l occurs at about 2 mg/day of lycopene from mixed dietary sources and a serum plateau occurs at >/=20 mg/day. CONCLUSIONS: These results show that lycopene and beta-carotene are bioavailable from red carrots and lycopene absorption seems to be affected by carrot fiber. Making inferences from both studies, the lycopene in the red carrot is about 44% as bioavailable as that from tomato paste. Red carrots provide an alternative to tomato paste as a good dietary source of lycopene and also provide bioavailable beta-carotene. PMID- 15116085 TI - Association between size at birth, truncal fat and obesity in adult life and its contribution to blood pressure and coronary heart disease; study in a high birth weight population. AB - OBJECTIVE: The aim of the study was to assess the relationship between size at birth and obesity as well as truncal fat, and its contribution to cardiovascular risk in a high birth weight population. DESIGN: Cohort-study with retrospectively collected data on size at birth. SETTING: Reykjavik, Iceland. SUBJECTS: A total of 1874 men and 1833 women born in Reykjavik during 1914-1935. MAIN OUTCOME MEASURES: Size at birth. Adult weight, height and skinfold thickness measurements, systolic and diastolic blood pressure, fatal and nonfatal coronary heart disease (CHD). RESULTS: Birth weight was positively related to adult body mass index (BMI) in both genders (B=0.35+/-0.14 kg/m(2), adj. R(2)=0.015, P=0.012 and B=0.34+/-0.17 kg/m(2), adj. R(2)=0.055, P=0.043 in men and women, respectively). However, high birth weight was not a risk factor for adult obesity (BMI>/=30 kg/m(2)). In the highest birth weight quartile, the odds ratio (95% CI) for being above the 90th percentile of truncal fat was 0.7 (0.6-1.0, P=0.021) for men and 0.4 (0.3-0.8, P=0.002) for women, compared with the lowest birth weight quartile. Truncal fat and BMI were positively related to blood pressure in both genders (P<0.05), but not to CHD. The regression coefficient for the inverse association between birth weight and blood pressure hardly changed when adding truncal fat to the model. CONCLUSION: In this high birth weight population, high birth weight was related to higher BMI in adulthood without being a risk factor for adult obesity. The inverse association between birth weight and truncal fat in adulthood suggests a role for foetal development in determining adult fat distribution. The inverse relationship of birth weight to blood pressure seems not to be mediated through the same pathway as to truncal fat. PMID- 15116086 TI - Effect of moderate changes in dietary fatty acid profile on postprandial lipaemia, haemostatic and related CVD risk factors in healthy men. AB - OBJECTIVE: To investigate the effect of moderate changes in dietary fatty acid profile on postprandial risk factors for cardiovascular disease (CVD). DESIGN: Double-blind, randomised, crossover, intervention trial. SETTING: : University of Auckland Human Nutrition Unit, New Zealand. SUBJECTS: A total of 18 lean healthy men. INTERVENTION: A dairy butter fat modified to reduce the saturated:unsaturated fatty acid ratio and a conventional high saturated butter fat were given on two separate occasions as a high-fat test meal (59+/-4 g fat; 71 en% fat) at breakfast. A fat exclusion lunch, dinner and snacks were also given. Blood samples were collected at 0 (baseline), 1, 3, 6, 10 and 24 h. RESULTS: Maximum peak in total triacylglycerol (TAG) occurred 3 h postprandially and was highest on modified treatment (diet, P<0.05) due predominantly to increased TAG within the chylomicron-rich fraction. Transient peaks in total-, LDL- and HDL-cholesterol occurred postprandially, but did not differ between dietary treatments (P>0.05). There were no differential effects of diet on postprandial free fatty acids, apo A, apo B, glucose, insulin, amylin or haemostatic clotting factors (P>0.05). CONCLUSIONS: In a group of healthy young men, replacement of 16% of total saturated fatty acids by mono- and polyunsaturated fats within a dairy lipid did not induce postprandial changes in CVD risk that may be considered beneficial for health. SPONSORSHIP: Fonterra, Wellington; New Zealand. PMID- 15116088 TI - The effect of docosahexaenoic acid-containing food administration on symptoms of attention-deficit/hyperactivity disorder-a placebo-controlled double-blind study. PMID- 15116087 TI - Patterns of physical activity in free-living adults in the Southern United States. AB - OBJECTIVE: To examine the relationship between the amount and patterns of physical activity (PA), body fatness, and age in a heterogeneous adult population in the free living. DESIGN: Cross-sectional study of the amount of PA over a 1 week period. The amount of body movements during PA (PA counts*10(3)) and time spent on various PA intensity categories were calculated from a triaxial accelerometer and compared with subject characteristics, including body fat from hydrodensitometry. PARTICIPANTS: Adult healthy men (n=48) and women (n=72) were recruited from the Nashville, Tennessee area and their PA was monitored in their free-living environment. RESULTS: The average weekday PA counts (176.5+/-60.3, P=0.002, r(2)=0.294), PA counts day-to-day variability (47.3+/-32.7, P=0.002, r(2)=0.286), daily maximum PA counts (241.9+/-89.2, P=0.001, r(2)=0.327), minute to-minute variability on weekdays (0.281+/-0.091, P=0.001, r(2)=0.362), and the difference between maximum and minimum daily PA counts (130.6+/-78.3, P=0.008, r(2)=0.243) were significantly and negatively correlated with body fatness. During awake time, both men and women spent 10-12 h on low intensity (1.0-2.9 metabolic equivalents (METs)) PA, approximately 1 h on moderate (3.0-5.9 MET), and less than 10 min on vigorous (>6.0 MET) PA each day. On weekends, men and women spent more time at rest (1 MET), less time on low-intensity PA, and men spent more time on moderate PA than on weekdays. CONCLUSIONS: In adults living in the Southern US the amount of free-living PA was negatively correlated with body fatness. Both men and women spent the majority of active time on low and moderate PA. PA patterns on weekends were different than on weekdays and were related to sex and age, but not to body fatness. SPONSORSHIP: National Institutes of Health, US. PMID- 15116089 TI - Blocking of FGFR signaling inhibits breast cancer cell proliferation through downregulation of D-type cyclins. AB - Overexpression of fibroblast growth factor receptor (FGFR) tyrosine kinases has been found in many human breast cancers and has been associated with poor patient prognosis. In order to understand the mechanism by which FGFR mediates breast cancer cell proliferation, we used a low molecular weight compound, PD173074, that selectively inhibits FGFR tyrosine kinase activity and autophosphorylation. This potential anticancer agent caused a G1 growth arrest of MDA-MB-415, MDA-MB 453 and SUM 52 breast cancer cells. Our analyses revealed that FGFR signaling links to the cell cycle machinery via D-type cyclins. PD173074-mediated inhibition of FGFR activity caused downregulation of cyclin D1 and cyclin D2 expression, inhibition of cyclin D/cdk4 activity and, as a consequence, reduction of pRB phosphorylation. Retroviral-mediated ectopic expression of cyclin D1 prevented pRB hypophosphorylation and the cell cycle G1 block in PD173074-treated cells, suggesting a central role for D cyclins in proliferation of FGFR-driven breast cancer cells. The repression of FGFR activity caused downregulation of MAPK in MDA-MB-415 and MDA-MB-453 cells. In SUM 52 cells, both MAPK and PI3K signaling pathways were suppressed. In conclusion, results shown here describe a mechanism by which FGFR promotes proliferation of breast cancer cells. PMID- 15116090 TI - Bone morphogenetic protein 3B silencing in non-small-cell lung cancer. AB - Bone morphogenetic protein 3B (BMP3B) is a member of the TGF-beta superfamily. The BMP3B promoter sequence was previously identified as a target for aberrant DNA methylation in non-small-cell lung cancer (NSCLC). Aberrant DNA hypermethylation in the BMP3B promoter is associated with downregulation of BMP3B transcription in both primary human lung cancers as well as lung cancer cell lines. In order to understand the mechanisms of BMP3B silencing in lung cancer, a sample set of 91 primary NSCLCs was used to detect aberrant BMP3B promoter methylation, mutations in the coding sequence of BMP3B, and loss of heterozygosity (LOH). Our results showed that 45 of 91 (or 49.5%) tested primary NSCLCs exhibited increased promoter methylation, and 40% demonstrated LOH in at least one of the flanking microsatellite markers sJRH and D10S196 (63 kb upstream or 3.338 Mbp downstream of BMP3B). The lung cancer cell line A549, a type II alveolar epithelial human lung cancer cell line, is characterized by aberrant DNA promoter methylation. We used retroviral vector constructs containing the BMP3B cDNA to re-express the gene in A549 cells and to investigate the effects on cell growth. No change in the cell growth rate was observed after BMP3B re-expression, as compared to the vector controls. Although the number of colonies formed in anchorage-dependent assays was only slightly decreased, the colony-forming ability of A549 cells after BMP3B expression in anchorage-independent assays in soft agar was significantly reduced to 10% (P<0.005, t-test). Moreover, the in vivo tumorigenicity assay in nude mice indicated that cells re-expressing BMP3B grew significantly slower than cells not expressing BMP3B (P<0.05, t-test). In conclusion, this study provides evidence that BMP3B expression is repressed by different mechanisms in lung cancer, and that the silencing of BMP3B promotes lung tumor development. PMID- 15116091 TI - Stat3 activation regulates the expression of matrix metalloproteinase-2 and tumor invasion and metastasis. AB - The expression of matrix metalloproteinase-2 (MMP-2) has been linked with tumor invasion, angiogenesis, and metastasis. However, the molecular basis for MMP-2 overexpression in tumor cells remains unclear. In this study, by using K-1735 melanoma system, we demonstrated that highly metastatic C4, M2, and X21 tumor cells express elevated MMP-2 mRNA and enzymatic activity, whereas poorly metastatic C10, C19, and C23 tumor cells express much lower levels. Moreover, a concomitant elevated Stat3 activity has been detected in these metastatic tumor cells that overexpress MMP-2. Transfection of constitutively activated Stat3 into poorly metastatic C23 tumor cells directly activated the MMP-2 promoter, whereas the expression of a dominant-negative Stat3 in highly metastatic C4 tumor cells inhibited the MMP-2 promoter. A high-affinity Stat3-binding element was identified in the MMP-2 promoter and Stat3 protein bound directly to the MMP-2 promoter. Blockade of activated Stat3 through expression of a dominant-negative Stat3 significantly suppressed MMP-2 expression in the metastatic tumor cells. Therefore, overexpression of MMP-2 in the metastatic melanoma cells can be attributed to elevated Stat3 activity, and Stat3 upregulates the transcription of MMP-2 through direct interaction with the MMP-2 promoter. Furthermore, blockade of activated Stat3 in highly metastatic C4 cells significantly suppressed the invasiveness of the tumor cells, inhibited tumor growth, and prevented metastasis in nude mice. Collectively, these studies suggest that Stat3 signaling directly regulates MMP-2 expression, tumor invasion, and metastasis, and that Stat3 activation might be a crucial event in the development of metastasis. PMID- 15116092 TI - Transgenic overexpression of Reg protein caused gastric cell proliferation and differentiation along parietal cell and chief cell lineages. AB - Reg (regenerating gene product) was originally identified as a growth factor involved in pancreatic regeneration. During the healing course of gastric erosion, Reg expression is highly increased in the enterochromaffin-like (ECL) cells surrounding the ulcer crater, suggesting its role as a regulator of gastric mucosal regeneration. However, there has been no direct in vivo evidence of a growth-promoting role of Reg for the gastric mucosal cells. In the current study, Reg-transgenic mice were created and gastric mucosa were analysed for histological changes. Transgenic mice showed a marked increase in the thickness of the fundic mucosa. Anti-proliferating cell nuclear antigen (PCNA) staining of the fundic mucosa demonstrated the enlargement of the proliferating neck zone and the lower PCNA-negative zone. Histological analysis employing antibodies against cell-type markers revealed expansion of the chief cell and parietal cell populations and no change in the number of surface mucus-producing cells, ECL cells, or G cells. In conclusion, Reg has a growth-promoting effect on gastric progenitor cells and an activity to direct the differentiation of the cells into chief cell and parietal cell lineages. This was in contrast to other factors, all of which had been shown to drive differentiation towards mucus producing cells in vivo. In the injured gastric mucosa, Reg may play a unique and important part in the reconstruction of the properly organized mucosal architecture. PMID- 15116093 TI - Phosphorylation of p53 on Thr55 by ERK2 is necessary for doxorubicin-induced p53 activation and cell death. AB - We recently reported that exposure of human cervical carcinoma cells to doxorubicin results in extracellular signal-regulated kinase (ERK)2 activation, which in turn phosphorylates p53 on a previously uncharacterized site, Thr55. This study sought to clarify the biological significance of doxorubicin-induced Thr55 phosphorylation. In breast carcinoma MCF7 cells, doxorubicin (300 nM) activated ERK2 and induced phosphorylation of p53 on Thr55 residues. Pretreatment of MCF7 cells with an ERK2 chemical inhibitor, PD98059 or U0126, blocked doxorubicin-induced p53 activation and suppressed phosphorylation of p53Thr55. MCF55a cells were established by transfection of full-length p53 carrying Thr55 mutation (Thr to Ala) into MCF7 cells. Doxorubicin (500 nM) could not induce p53 activation in MCF55a cells, which showed significantly increased drug resistance toward doxorubicin. While the expression of the apoptotic protein, Bax, showed no difference between MCF7 and MCF55a cells, Bcl-2, an antiapoptotic protein, was constitutively expressed in MCF55a cells. The increase of Bcl-2 protein and/or Bcl-2/Bax ratio might at least partly contribute to the drug resistance of MCF55a cells. In summary, our results suggest that phosphorylation of p53Thr55 by ERK2 is important for doxorubicin-induced p53 activation and cell death. PMID- 15116094 TI - Disruption of 14-3-3 binding does not impair Protein 4.1B growth suppression. AB - Meningiomas are common central nervous system tumors; however, the mechanisms underlying their pathogenesis are largely unknown. Collaborative studies from our laboratory demonstrated a direct association of 14-3-3 with the meningioma tumor suppressor Protein 4.1B, which was not observed with other members of the Protein 4.1 family, including the NF2 meningioma tumor suppressor, merlin/schwannomin. Given the role of 14-3-3 in the regulation of cell proliferation and apoptosis, we sought to determine the functional significance of 14-3-3 binding to Protein 4.1B growth suppression. Based on comparative binding studies performed with additional members of the Protein 4.1 family, we generated specific missense mutations within the minimal growth suppressor fragment of Protein 4.1B (DAL-1, differentially expressed in adenocarcinoma of the lung). Complementary in vitro GST affinity chromatography and in vivo interaction experiments demonstrated that the F359Y mutation abrogated binding to 14-3-3, but did not impair DAL-1 binding to other known Protein 4.1B interacting proteins. Similar to wild-type DAL-1, the expression of the F359Y DAL-1 14-3-3-binding mutant resulted in reduced Protein 4.1B-deficient IOMM-Lee and CH157-MN meningioma cell line colony formation. Moreover, similar to wild-type DAL-1, the stable expression of the DAL-1 F359Y mutant significantly reduced cell proliferation in independently isolated IOMM Lee clones, as assessed by thymidine incorporation. Collectively, these results suggest that binding to 14-3-3 is not essential for the growth suppressor function of Protein 4.1B in meningiomas. PMID- 15116095 TI - Arsenic inhibition of the JAK-STAT pathway. AB - The Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway is an essential cascade for mediating normal functions of different cytokines in the development of the hematopoietic and immune systems. Chronic exposure to arsenic has been found to cause immunotoxicity and has been associated with the suppression of hematopoiesis (anemia and leukopenia). Here, we report the novel finding of arsenic-mediated inactivation of the JAK-STAT signaling pathway by its direct interaction with JAK tyrosine kinase. Pretreatment with sodium arsenite strongly inhibited IL-6-inducible STAT3 tyrosine phosphorylation in HepG2 cells and did not affect its serine phosphorylation. As a result, sodium arsenite completely abolished STAT activity dependent expression of suppressors of cytokine signaling (SOCS). Both cellular and subcelluar experiments showed that the inhibition of JAK-STAT signaling resulted from JAK tyrosine kinase's direct interaction with arsenite, and that arsenic's suppression of JAK tyrosine kinase activity also occurred in the interferon gamma (IFNgamma) pathway. The ligand-independent inhibition by arsenic indicates that JAK was the direct target of arsenic action. Other inflammatory stimulants, stress agents, and metal cadmium failed to induce similar effects on the tyrosine phosphorylation of STAT3 as arsenic does. Our experiments also revealed that arsenic inactivation of the JAK-STAT pathway occurred independent of arsenic activation of MAP kinases. Taken together, our findings indicate that arsenic directly inhibits JAK tyrosine kinase activity and suggest that this direct interference in the JAK-STAT pathway may play a role in arsenic-associated pathogenesis. PMID- 15116096 TI - Enhanced tenascin-C expression and matrix deposition during Ras/TGF-beta-induced progression of mammary tumor cells. AB - Overexpression of tenascin-C (TN-C) in breast carcinomas has been associated with a migratory or even invasive tumor cell phenotype. The mechanisms regulating expression and matrix deposition of TN-C in normal and cancerous breast tissues are, however, little understood. Here, we demonstrate that mouse mammary epithelial cells (EpH4) transformed by oncogenic Ha-Ras (EpRas) overexpress TN-C, which accumulates in the cytoplasm. When EpRas cells undergo epithelial mesenchymal transition (EMT) in response to TGFbeta1, they secrete TN-C into the culture medium. In EpRas cells undergoing TGFbeta1-induced EMT in three dimensional (3D)-collagen gel cultures, TN-C was deposited into an extracellular matrix (ECM) already containing fibronectin and perlecan. Under less physiological 2D plastic cultures, EpRas cells undergoing EMT failed to deposit TN-C into an (apparently incomplete) ECM. Ras-downstream signaling was dissected by pharmacological inhibitors and effector-specific Ras mutants (V12S35, V12C40), specifically inhibiting or activating ERK/MAPK or PI3K signaling, respectively. We showed that TN-C overexpression required a hyperactive ERK/MAPK-signaling pathway, while elevated PI3K signaling did not enhance TN-C expression. Similarly, tumors induced by cells exhibiting hyperactive ERK/MAPK signaling showed expression of TN-C in the tumor cells themselves, while only endothelial cells expressed TN-C in tumors caused by the V12C40 mutant (incapable of EMT in vivo). Taken together, our data indicate that hyperactive ERK/MAPK signaling causes enhanced expression of TN-C, while its secretion is induced by TGFbeta1 and both signals cooperate in TN-C matrix deposition. Importantly, both signals also cooperate to induce EMT in vitro and tumor progression/metastasis in vivo. PMID- 15116097 TI - A negative role of SHP-2 tyrosine phosphatase in growth factor-dependent hematopoietic cell survival. AB - SHP-2 tyrosine phosphatase is highly expressed in hematopoietic cells; however, the function of SHP-2 in hematopoietic cell processes is not fully understood. Recent identification of SHP-2 mutations in childhood leukemia further emphasizes the importance of SHP-2 regulation in hematopoietic cells. We previously reported that SHP-2 played a positive role in IL-3-induced activation of Jak2 kinase in a catalytic-dependent manner. Interestingly, enforced expression of wild-type (WT) SHP-2 in Ba/F3 cells enhanced growth factor deprivation-induced apoptosis. Biochemical analyses revealed that although IL-3 activation of Jak2 kinase was increased, tyrosyl phosphorylation of its downstream substrate STAT5 was disproportionately decreased by the overexpression of SHP-2. Following IL-3 deprivation, the tyrosyl phosphorylation of STAT5 that is required for its antiapoptotic activity was rapidly diminished in SHP-2 overexpressing cells. As a result, reduction of the putative downstream targets of STAT5-Bcl-X(L) and pim-1 was accelerated by overexpression of SHP-2. Further investigation showed that SHP 2 associated with STAT5, and that it was indeed able to dephosphorylate STAT5. Finally, overexpression of SHP-2 in primary bone marrow hematopoietic progenitor cells compromised their differentiative and proliferative potential, and enhanced growth factor withdrawal-induced cell death. And, the effect of SHP-2 overexpression on growth factor-dependent survival was diminished in STAT5 deficient hematopoietic cells. Taken together, these results suggest that SHP-2 tyrosine phosphatase negatively regulates hematopoietic cell survival by dephosphorylation of STAT5. PMID- 15116098 TI - Cancer predisposition in mice deficient for the metastasis-associated Mts1(S100A4) gene. AB - Metastasis-promoting Mts1(S100A4) protein belongs to the S100 family of Ca(2+) binding proteins. A mouse strain with a germ-line inactivation of the S100A4 gene was generated. The mice were viable and did not display developmental abnormalities in the postnatal period. However, an abnormal sex ratio was observed in the litters with the S100A4-/- genotype, raising the possibility of a certain level of embryonic lethality in this strain. In all, 10% of 10-14-month old S100A4-null animals developed tumors. This is a characteristic feature of mouse strains with inactivated tumor suppressor genes. Spontaneous tumors of S100A4-/- mice were p53 positive. Recently, we have shown that S100A4 interacts with p53 tumor suppressor protein and induces apoptosis. We proposed that impairment of this interaction could affect the apoptosis-promoting function of p53 that is involved in its tumor suppressor activity. The frequency of apoptosis in the spleen of S100A4-/- animals after whole-body gamma-irradiation was reduced compared to the wild-type animals. The same was true for the transcriptional activation of the p53 target genes - waf/p21/cip1 and bax. Taken together, these observations indicate that spontaneous tumors in S100A4-/- mice are a result of functional destabilization of p53 tumor suppressor gene. PMID- 15116099 TI - HST-1/FGF-4 plays a critical role in crypt cell survival and facilitates epithelial cell restitution and proliferation. AB - The fibroblast growth factor-4 (HST-1/FGF-4) is a heparin-binding growth factor that influences on epithelial and many other cells through interaction with FGF receptors. It has been demonstrated that the HST-1/FGF-4 gene protects mice from lethal irradiation by preventing bone marrow damage and intestinal tract damage. However, the radioprotective mechanism is unknown. In this study, we have investigated the expression of Hst-1/Fgf-4 in mouse small intestine after irradiation, and determined the role of HST-1/FGF-4 in mouse intestinal crypt cell survival and epithelial cell proliferation and restitution. We found the induction of endogenous Hst-1/Fgf-4 expression in intestine when mice are exposed to 9.0 Gy irradiation. Laser-captured microdissection (LCM) coupled with RT-PCR analysis revealed that expression of Hst-1/Fgf-4 was found in epithelial cell of the villi and crypt cells. Pretreatment of HST-1/FGF-4 caused an increase in the number of surviving crypt cells, and clearly suppresses the radiation-induced apoptosis of the crypt cells. Moreover, exogenous HST-1/FGF-4 enhances epithelial cell restitution and proliferation in an in vitro model. These data suggest that HST-1/FGF-4 is induced by irradiation injury, and that HST-1/FGF-4 will find a therapeutic role in the prevention of intestinal cell toxicity following intensive chemotherapy and radiation therapy protocols, and in allogenic cell transplantation. PMID- 15116100 TI - Gastrin-mediated activation of cyclin D1 transcription involves beta-catenin and CREB pathways in gastric cancer cells. AB - Gastrin and its precursors promote proliferation in different gastrointestinal cells. Since mature, amidated gastrin (G-17) can induce cyclin D1, we determined whether G-17-mediated induction of cyclin D1 transcription involved Wnt signaling and CRE-binding protein (CREB) pathways. Our studies indicate that G-17 induces protein, mRNA expression and transcription of the G(1)-specific marker cyclin D1, in the gastric adenocarcinoma cell line AGSE (expressing the gastrin/cholecystokinin B receptor). This was associated with an increase in steady-state levels of total and nonphospho beta-catenin and its nuclear translocation, indicating the activation of the Wnt-signaling pathway. In addition, G-17-mediated increase in cyclin D1 transcription was significantly attenuated by axin or dominant-negative (dn) T-cell factor 4(TCF4), suggesting crosstalk of G-17 with the Wnt-signaling pathway. Mutational analysis indicated that this effect was mediated through the cyclic AMP response element (CRE) (predominantly) and the TCF sites in the cyclin D1 promoter, which was also inhibited by dnCREB. Furthermore, G-17 stimulation resulted in increased CRE responsive reporter activity and CREB phosphorylation, indicating an activation of CREB. Chromatin immunoprecipitation studies revealed a G-17-mediated increase in the interaction of beta-catenin with cyclin D1 CRE, which was attenuated by dnTCF4 and dnCREB. These results indicate that G-17 induces cyclin D1 transcription, via the activation of beta-catenin and CREB pathways. PMID- 15116101 TI - cDNA cloning and characterization of a novel gene encoding the MLF1-interacting protein MLF1IP. AB - Myelodysplasia/acute myeloid leukemia (MDS/AML) is characterized by a t(3;5)(q25.1;q34) chromosomal translocation that forms a fusion gene between nucleophosmin (NPM) and MDS/myeloid leukemia factor 1 (MLF1). We identified a novel protein, MLF1-interacting protein (MLF1IP), that specifically associates with MLF1 by yeast two-hybrid analysis and in pulldown assays, and colocalizes with it in both the nuclei and cytoplasm of cells. The MLF1IP gene locus is at chromosome 4q35.1 and is composed of 14 exons spanning 75.8 kb of genomic DNA. The MLF1IP cDNA encodes a 46-kDa protein that contains two bipartite and two classical nuclear localization signals, two nuclear receptor-binding motifs (LXXLL), two leucine zippers, two PEST residues and several potential phosphorylation sites. MLF1IP transcripts are expressed in a variety of tissues (e.g. fetal liver, bone marrow, thymus and testis). MLF1IP appears to be a lineage-specific gene whose expression is confined exclusively to the CFU-E erythroid precursor cells, but not in mature erythrocytes. These observations, together with previous data demonstrating a role for MLF1 in suppressing red cell maturation, suggest a possible role for MLF1IP and MLF1 deregulation in the genesis of erythroleukemias. PMID- 15116102 TI - Expression of CD109 in human cancer. AB - It was recently reported that the human CD109 gene encodes a glycosyl phosphatidylinositol-anchored glycoprotein that is a member of the alpha(2) macroglobulin/C3, C4, C5 family of thioester-containing proteins. In this study, we found that the expression of mouse CD109 gene was upregulated in NIH3T3 cells expressing RET tyrosine kinase with a multiple endocrine neoplasia 2B mutation. Northern blot analysis showed a high level of expression of the CD109 gene only in the testis in normal human and mouse tissues. In addition, its expression was high in some human tumor cell lines, which included squamous cell carcinoma and glioblastoma cell lines, whereas it was undetectable in neuroblastoma and small cell lung carcinoma cell lines. When CD109 expression was examined in 33 cases of human lung cell carcinomas by quantitative RT-PCR, a significant high expression of CD109 was detected in about half of squamous cell carcinomas examined, but not in adenocarcinoma, large-cell carcinoma and small-cell carcinoma. Similarly, upregulation of CD109 was observed in nine out of 17 esophageal squamous cell carcinomas. Thus, these results suggested that CD109 might be a useful molecular target for the development of new therapeutics for malignant tumors, such as squamous cell carcinoma. PMID- 15116103 TI - p73-alpha is capable of inducing scotin and ER stress. AB - p73, like its family member p53, can induce programmed cell death following DNA damage. Here, we report that this mechanism also involves endoplasmic reticulum (ER) stress and the transactivation of scotin, a protein identified recently as a p53 target able to induce ER stress. By using Tet-On inducible cell lines (Saos 2 osteosarcoma cells that lack p53), we observed that TAp73alpha elicits significant alterations in the morphology of the ER system, namely in the fine subcellular localization of calnexin. We found that both TAp73alpha and p53 are strong inducers of scotin. On the other hand, the transcriptionally deficient short isoforms DeltaNp73alpha did not upregulate the steady-state mRNA level of scotin, as evaluated by real-time RT-PCR. Following the induction of scotin, ER staining with calnexin showed evidence of morphological alteration, with variations in the intracellular concentration of free calcium, visualized by fluo 3 staining. The induction of ER stress by p73 was further supported by the transcriptional induction of Gadd 153, a transcription factor induced under ER stress conditions. In conclusion, the data reported indicate the ability of TAp73alpha and p53 (not DeltaNp73alpha) to elicit scotin transactivation and ER stress. This molecular mechanism might contribute to the effector events inducing apoptosis downstream of p73. PMID- 15116104 TI - p38 MAP kinase signaling is necessary for rat chondrosarcoma cell proliferation. AB - Chondrosarcomas represent the second most frequent class of primary skeletal malignancies. This tumor type is highly resistant to radiation therapy and currently available chemotherapies, thereby limiting treatment choice to surgical resection. Identifying the mechanisms responsible for chondrosarcoma cell proliferation is therefore crucial for the development of new treatment strategies. Here, we demonstrate a significant reduction in rat chondrosarcoma cell proliferation following treatment with pharmacological inhibitors (SB202190 and PD169316) of p38 mitogen-activated protein (MAP) kinases. In an attempt to dissect possible mechanisms, we investigated the effect of p38 inhibition on promoter activity of cell-cycle genes. Surprisingly, p38 inhibition resulted in upregulation of the activities of all three D-type cyclin promoters. In addition, p38 inhibitors induced increased transcription of the cell-cycle inhibitor p21(waf1/cip1). As expected, promoter activity of the cyclin A gene, which lies downstream of D-type cyclins and p21 in cell-cycle progression, was strongly reduced by p38 inhibitors. These effects were independent of a cyclic AMP response element and conferred by the proximal 150 nucleotides of the cyclin A promoter. Decreased transcription was accompanied by greatly reduced cyclin A protein levels upon p38 inhibition. These observations indicate complex regulation of chondrosarcoma cell-cycle progression by p38 signaling, and suggest that components of p38 MAP kinase pathways may be effective targets in the treatment of these tumors. PMID- 15116105 TI - Help wanted--to immunize the world. PMID- 15116106 TI - Australians say yes to more research funding. PMID- 15116107 TI - A healthier climate for the funding of vaccine research. AB - Vaccination is a marvel of scientific endeavor that benefits the masses. Yet the laissez-faire economy may not provide a sufficient push for vaccine research and development. The current climate that drives this globally important venture is examined here. PMID- 15116108 TI - Vaccine development strategies for improving immunization: the role of modern immunology. AB - An ideal vaccine has certain biological and physical characteristics. Technological advances have provided new strategies that may help the design of such a vaccine. PMID- 15116109 TI - Ethics of vaccine research. AB - Vaccination has attracted controversy at every stage of its development and use. Ethical debates should consider its basic goal, which is to benefit the community at large rather than the individual. PMID- 15116110 TI - Resurrecting CD8+ suppressor T cells. PMID- 15116111 TI - Dissecting worm immunity. PMID- 15116112 TI - Toll signaling: RIPping off the TNF pathway. PMID- 15116113 TI - Nature's fortress against infection. PMID- 15116114 TI - Paradigm switching in the germinal center. PMID- 15116117 TI - An autonomous molecular computer for logical control of gene expression. AB - Early biomolecular computer research focused on laboratory-scale, human-operated computers for complex computational problems. Recently, simple molecular-scale autonomous programmable computers were demonstrated allowing both input and output information to be in molecular form. Such computers, using biological molecules as input data and biologically active molecules as outputs, could produce a system for 'logical' control of biological processes. Here we describe an autonomous biomolecular computer that, at least in vitro, logically analyses the levels of messenger RNA species, and in response produces a molecule capable of affecting levels of gene expression. The computer operates at a concentration of close to a trillion computers per microlitre and consists of three programmable modules: a computation module, that is, a stochastic molecular automaton; an input module, by which specific mRNA levels or point mutations regulate software molecule concentrations, and hence automaton transition probabilities; and an output module, capable of controlled release of a short single-stranded DNA molecule. This approach might be applied in vivo to biochemical sensing, genetic engineering and even medical diagnosis and treatment. As a proof of principle we programmed the computer to identify and analyse mRNA of disease-related genes associated with models of small-cell lung cancer and prostate cancer, and to produce a single-stranded DNA molecule modelled after an anticancer drug. PMID- 15116118 TI - Generation of B-cell chronic lymphocytic leukemia (B-CLL)-reactive T-cell lines and clones from HLA class I-matched donors using modified B-CLL cells as stimulators: implications for adoptive immunotherapy. AB - Allogeneic stem cell transplantation following reduced-intensity conditioning is being evaluated in patients with advanced B-cell chronic lymphocytic leukemia (B CLL). The curative potential of this procedure is mediated by donor-derived alloreactive T cells, resulting in a graft-versus-leukemia effect. However, B-CLL may escape T-cell-mediated immune reactivity since these cells lack expression of costimulatory molecules. We examined the most optimal method to transform B-CLL cells into efficient antigen-presenting cells (APC) using activating cytokines, by triggering toll-like receptors (TLRs) using microbial pathogens and by CD40 stimulation with CD40L-transfected fibroblasts. CD40 activation in the presence of IL-4 induced strongest upregulation of costimulatory and adhesion molecules on B-CLL cells and induced the production of high amounts of IL-12 by the leukemic cells. In contrast to primary B-CLL cells as stimulator cells, these malignant APCs were capable of inducing the generation of B-CLL-reactive CD8(+) CTL lines and clones from HLA class I-matched donors. These CTL lines and clones recognized and killed primary B-CLL as well as patient-derived lymphoblasts, but not donor cells. These results show the feasibility of ex vivo generation of B-CLL-reactive CD8(+) CTLs. This opens new perspectives for adoptive immunotherapy, following allogeneic stem cell transplantation in patients with advanced B-CLL. PMID- 15116119 TI - Acute promyelocytic leukemia cell line AP-1060 established as a cytokine dependent culture from a patient clinically resistant to all-trans retinoic acid and arsenic trioxide. AB - AP-1060 is a newly established acute promyelocytic leukemia (APL) cell line from a multiple-relapse patient clinically resistant to both all-trans retinoic acid (ATRA) and arsenic trioxide (ATO). The line was initially derived as a granulocyte colony-stimulating factor-dependent strain that underwent replicative senescence and, following ethylnitrosourea treatment, as a phenotypically similar immortalized line. Immortalization was associated with broadened cytokine sensitivity but not growth autonomy, in contrast to three previously derived APL lines. Both the AP-1060 strain and line had shortened telomeres and low telomerase activity, while the line had higher expression of many genes associated with macromolecular synthesis. The karyotype was 46,XY,t(3;14)(p21.1;q11.2),t(15;17)(q22;q11)[100%]; the unique t(3;14) was observed in 4/9 t(15;17)-positive metaphase cells at previous relapse on ATRA therapy. The PML-RARalpha mRNA harbored a missense mutation in the RARalpha region ligand-binding domain (Pro900Ser). This was associated with a right-shift and sharpening of the ATRA-induced maturation response compared to ATRA-sensitive NB4 cells, which corresponded to the transcriptional activation by PML RARalphaPro900Ser of a cotransfected ATRA-targeted reporter vector in COS-1 cells. AP-1060 also manifested relative resistance to ATO-induced apoptosis at >/=1 microM, while 0.25 microM ATO stimulated limited atypical maturation. These findings suggest that AP-1060 will be useful for further assessing molecular elements involved in APL progression and drug response/resistance. PMID- 15116120 TI - Expression of APO2.7, bcl-2 and bax apoptosis-associated proteins in CD34- bone marrow cell compartments from patients with myelodysplastic syndromes. PMID- 15116121 TI - Topoisomerase IIalpha expression in mantle cell lymphoma: a marker of cell proliferation and a prognostic factor for clinical outcome. AB - Mantle cell lymphoma (MCL) is a malignant lymphoma associated with a relatively aggressive clinical course and a median overall survival time of 3-4 years. Treatment usually consists of combination chemotherapy, often including topoisomerase (topo) inhibitors such as doxorubicin, etoposide and mitoxantrone. Topo IIalpha is an enzyme that is needed whenever uncoiling of DNA is necessary during the cell cycle. The enzyme is a marker of cell proliferation. We analyzed the expression of topo IIalpha in relation to Ki-67 and the clinical outcome in patients with MCL. Biopsy specimens from 95 untreated patients enrolled in two multicenter trials (1975-1985) were investigated immunohistochemically with monoclonal antibodies against topo IIalpha (Ki-S4) and Ki-67 (Ki-S5). Patients with low (0-10%) topo IIalpha expression had a median overall survival time of 49.0 months, compared to 17.0 months for patients with high (more than 10%) topo IIalpha expression. The Kaplan-Meier analysis showed a significant difference in the overall survival time related to the percentage of topo IIalpha (P<0.001) and Ki-67 (P<0.001) positive tumor cells. Multivariate Cox regression analysis revealed the expression of topo IIalpha as the most important prognostic factor (P<0.001) in MCL superior to the international prognostic index (IPI), the Ki-67 index and other clinical characteristics. PMID- 15116123 TI - Valproic acid inhibits proliferation and induces apoptosis in acute myeloid leukemia cells expressing P-gp and MRP1. AB - The multidrug resistance (MDR) phenotype, induced by the overexpression of several ABC transporters or by antiapoptotic mechanisms, has been identified as the major cause of drug resistance in the treatment of patients with acute myeloid leukemia (AML). In this study, we have shown that valproic acid (VPA) (a histone deacetylase inhibitor) can inhibit the proliferation of both P glycoprotein (P-gp)- and MDR-associated protein 1 (MRP1)-positive and -negative cells. VPA also induced apoptosis of P-gp-positive cells. VPA induced apoptosis in K562 cells led to decrease in Flip (FLICE/caspase-8 inhibitory protein) expression with Flip cleavage, which could not be observed in HL60 cells. In HL60/MRP cell line, which proved to be resistant to apoptosis by VPA, we observed an abnormal expression of apoptotic regulatory proteins, overexpression of Bcl-2 and absence of Bax. Also, the Bcl-2 antagonist HA14-1 rapidly restored apoptosis in this cell line. Cotreatment with cytosine arabinoside induced very strong apoptosis in both K562/DOX and HL60/DNR cell lines. VPA also induced apoptosis in AML patient cells expressing P-gp and/or MRP1. Our findings show VPA as an interesting drug that should be tested in clinical trials for overcoming the MDR phenotype in AML patients. PMID- 15116122 TI - The histone deacetylase inhibitor MS-275 induces caspase-dependent apoptosis in B cell chronic lymphocytic leukemia cells. AB - MS-275 is a histone deacetylase (HDAC) inhibitor that has been reported to mediate its cytotoxic effect through generation of reactive oxygen species (ROS) in proliferating hematopoietic cell lines. We examined efficacy of MS-275 in nonproliferating chronic lymphocytic leukemia (CLL) cells from patients. In these cells, MS-275 demonstrated an in vitro LC(50) that was one log lower than for normal mononuclear cells. Following MS-275 treatment, histones H3 and H4 showed increased acetylation and HDAC enzymatic activity was reduced. Caspase-8, -9, and -3 were activated, and caspase substrates PARP and BID were cleaved. Additionally, FLICE-inhibitory protein (FLIP) was downmodulated following MS-275 incubation. MS-275 treatment caused detectable ROS generation after 15 h of incubation, which was blocked by the caspase inhibitor Z-VAD-fmk. Overexpression of Bcl-2 protein protected against MS-275-induced apoptosis. These data demonstrate that MS-275 is a promising therapy for the treatment of CLL, but that in contrast to previous reports, ROS generation does not precede commitment to apoptosis. Similar to many other therapeutic targets, MS-275-mediated apoptosis is reduced by overexpression of Bcl-2, justifying strategies to combine HDAC inhibitors with Bcl-2 antagonists. PMID- 15116124 TI - Optimizing long-term care by administration of influenza vaccine to parents of NICU patients. AB - The neonatal intensive care unit (NICU) team has traditionally invested itself in maintaining the health of its patients upon discharge from high-acuity care. Historically, this has included the administration of vaccinations to the patients and more recently, Palivizumab--a monoclonal antibody directed against respiratory synctial virus (RSV). With increasing awareness of the ill-effects associated with influenza virus and recommendations those in close contact with high-risk infants receive the vaccine, the NICU may be an ideal arena to capture parents of high-risk infants for vaccination. This would potentially decrease exposure of the neonatal patient group to influenza virus and may decrease morbidity and mortality associated with the disease. NICUs should work in concert with their associated Departments of Obstetrics to immunize pregnant mothers when appropriate, educate parents regarding influenza and its potential effects in infants and offer influenza vaccine in-season to parents as part of comprehensive care. PMID- 15116125 TI - Use of recombinant factor VIIa in infants with severe coagulopathy. AB - The risk of hemorrhage in infants with severe coagulopathies unresponsive to fresh frozen plasma (FFP) infusions may preclude therapeutic invasive interventional procedures. We describe the successful use of recombinant factor VIIa (rFVIIa) in two such infants, the first with cirrhosis requiring paracentesis and the second with necrotizing enterocolitis requiring laparotomy. This report reviews the current concepts on the mechanism of action of the drug rFVIIa and considers its expanded use in infants unresponsive to FFP replacement. PMID- 15116126 TI - Chloride channel CICN7 mutations in a Korean patient with infantile malignant osteopetrosis initially presenting with neonatal thrombocytopenia. PMID- 15116127 TI - Serendipitous diagnosis of infracardiac total anomalous pulmonary venous return by umbilical venous catheter blood gas samples. AB - Total anomalous pulmonary venous return (TAPVR) is a rare congenital heart defect that occurs when all four pulmonary veins connect to the systemic venous circulation. We describe a full-term male neonate who presented with cyanosis and mild tachypnea shortly after birth. One umbilical artery and the umbilical vein were catheterized, and oxygen treatment was provided. Four echocardiograms indicated severe pulmonary hypertension and were negative for any congenital heart defects. After the umbilical artery catheter was removed, high partial pressure of oxygen was detected in blood samples drawn from the umbilical venous catheter that was positioned below the diaphragm. Based on this finding, TAPVR was suspected and confirmed with angiography through a central venous catheter. The neonate underwent a successful surgical repair to correct the cardiac defect. PMID- 15116128 TI - Intramural bronchogenic cyst in the carina observed in a neonate and treated by needle aspiration: a case report. AB - We report the first case to be observed in a neonate of an intramural bronchogenic cyst in the carina. Considering the age of the infant, it was decided to administer curative treatment by needle aspiration. A rigid bronchoscopy was used. The outcome was favorable. PMID- 15116129 TI - A rare cause of intestinal perforation in an extreme low birth weight infant- gastrointestinal mucormycosis: a case report. AB - We report an extreme low birth weight infant with gastrointestinal mucormycosis (GM) mimicking necrotizing enterocolitis on day 15 of age. Emergency laparotomy confirmed ileal perforations. Subsequently, tissue histology confirmed GM. This is the most premature infant who survived after GM and is the third reported neonatal survivor in English literature. PMID- 15116130 TI - The Fanconi-Bickel syndrome: a case of neonatal onset. AB - A male newborn infant was recognized having Fanconi-Bickel syndrome (FBS) in the neonatal period. The presenting clinical findings were hyperglycemia and polyuria detected during an episode of acute enteritis. Physical examination was normal, biochemical analyses were suggestive of FBS: glycosuria, proteinuria, phosphaturia, generalized aminoaciduria, and increased levels of urinary beta 2 microglobulin, serum glucose and serum alkaline phosphatase. The molecular genetic analysis showed homozygosity for mutations within the gene of the glucose transporter 2 (Glut 2), 1213 C>T. The patient demonstrated improved clinical and metabolic status following institution of diet with frequent small meals and galactose-free-milk as well as pharmacological treatment with phosphate and vitamin alpha-OH-D3. In conclusion, infants showing hyperglycemia and polyuria may be considered having FBS also in the neonatal period. Early institution of adequate caloric intake and replacement of electrolytes and vitamin D may avoid or reduce metabolic complications. PMID- 15116131 TI - Mixed infection with unusual fungi and staphylococcal species in two extremely premature neonates. AB - Systemic fungal infections are being recognized with increasing frequency in extremely premature neonates. We report two such infants with late-onset mixed infection with Staphylococcal species and unusual fungi. These cases are of interest in view of recent reports on the interaction of Staphylococcal cell wall components and neutrophils, as damaged skin sites could form a nidus and portal of entry for saprophytic fungal pathogens. It is also important to consider fungal infection as a possibility in sick premature infants with necrotic skin lesions even when the systemic signs have an alternative explanation with ongoing bacteremia. PMID- 15116132 TI - Haemangioma presenting as perianal ulcer in neonates. AB - Perianal haemangiomas are not uncommon, however, their presentation in the neonate as an ulcer prior to the development of a clinical haemangioma is extremely rare. Two such cases are reported. PMID- 15116133 TI - Sirenomelia with an angiomatous lumbosacral myelocystocele in a full-term infant. AB - Sirenomelia, also known as the mermaid syndrome, is a rare congenital malformation of uncertain etiology. It is characterized by fusion of the lower limbs and commonly associated with severe urogenital and gastrointestinal malformations. In this report, we describe the first case of an infant with sirenomelia and a massive angiomatous lumbosacral myelocystocele. PMID- 15116135 TI - Commentary regarding attempt to obtain hyperbaric oxygen treatment and the decision to sue is poorly researched and inaccurate. PMID- 15116137 TI - Predicting outcome in neonatal hypoxic respiratory failure with the score for neonatal acute physiology (SNAP) and highest oxygen index (OI) in the first 24 hours of admission. AB - OBJECTIVE: To determine the clinical utility of SNAP score versus the highest oxygen index (OI) in first 24 hours of admission in predicting outcome of HRF. STUDY DESIGN: All admissions (1991 to 1999) > or =36 weeks gestation, ventilated for > or =12 hours with FiO(2)> or =0.50, without congenital anomalies were reviewed. Primary outcome measure was survival (without ECMO) versus ECMO and/or death. RESULTS: From 184 infants with HRF, 148 survived (without ECMO) versus 36 died and/or received ECMO. SNAP score and highest OI were similar in predicting outcome of HRF (area under ROC curve: 0.813+/-0.037 versus 0.814+/-0.041; P=0.72). Death and/or ECMO requirement were best predicted by a SNAP score of 19 (Sensitivity 75.0%, Specificity 71%) or an OI of 28 (Sensitivity 75.0%, Specificity 76.4%). CONCLUSION: Although both, the SNAP score and highest OI, are useful and similar in predicting outcome of HRF, OI is preferable because of its ease of use. We believe the predictive value of these parameters should be evaluated in a multicenter setting. PMID- 15116138 TI - Hospital survival of very-low-birth-weight neonates from 1977 to 2000. AB - OBJECTIVE: To determine patterns of survival for very low birth weight (VLBW, birth weight 501 to 1500 g) neonates over 23 years. STUDY DESIGN: Data for 4873 VLBW neonates born from 1977 to 2000 were divided into five epochs. The primary outcome was survival to hospital discharge. Birth weight-specific survival rates were estimated by race and gender for each epoch. Presence of comorbidities and congenital anomalies, delivery mode, and provision of artificial ventilation were investigated to determine whether they could explain observed survival patterns. RESULTS: From 1977 to 1995, survival increased from 50.2% to 81.0% as the proportion of VLBW neonates receiving artificial ventilation rose from 59.0% to 80.9%. Survival was unchanged between 1990 to 1995 and 1996 to 2000. Black females maintained a survival advantage over the entire study period. Survival improved for neonates with congenital anomalies over time, but had little impact on race/gender survival patterns. Survival patterns also could not be explained by comorbidity status, delivery mode, or access to artificial ventilation. CONCLUSION: The survival advantage of VLBW black females persists and remains unexplained. PMID- 15116139 TI - Increased morbidity in severe early intrauterine growth restriction. AB - OBJECTIVE: To determine the relative frequencies of complications in severe early intrauterine growth-restricted (IUGR) infants. METHODS: All infants 32 weeks gestation or less with birth weight less than the fifth percentile admitted from January 1991 to December 1998 were identified retrospectively. Two infants were identified for each IUGR case: the subsequent admission with birth weight +/-100 g of the case, and the subsequent admission with the same gestational age. Infants with multiple congenital anomalies, congenital infections or admission after 48 hours of age were excluded. Maternal and neonatal demographic data, neonatal morbidity and mortality until discharge were gathered by chart review. RESULTS: A total of 39 IUGR identified infants met criteria, with 41 gestational age infants and 33 birth weight infants. Mean birth weights and gestational ages for the IUGR group, gestational age group, and birth weight group were 744 g and 29.6 weeks, 1370 g and 29.7 weeks, and 781 g and 25.5 weeks respectively. Mortality was higher for IUGR infants than gestational age infants (20.5 vs 0%), but less than the birth weight infants (30%). In surviving infants, total ventilator days, total oxygen days, days to full feeds, and patent ductus arteriosis, were higher for IUGR infants than gestational age infants, but less than birth weight infants. Hypoglycemia, direct hyperbilirubinemia, necrotizing enterocolitis (NEC), thrombo-cytopenia, chronic lung disease and feeding difficulties occurred more frequently in IUGR infants than in both other groups. Length of stay for survivors and incidence of retinopathy of prematurity (ROP) was similar for the IUGR and birth weight infants. CONCLUSIONS: Infants born prematurely who are also severely IUGR have higher neonatal morbidity and mortality when compared to infants of similar gestational age. The surviving IUGR infants had less intraventricular hemorrhage and periventricular leukomalacia than less mature infants of comparable birth weight, but a similar incidence of ROP and length of stay. They had a higher incidence of NEC, direct hyperbilirubinemia and chronic lung disease, probably due to end-organ damage in utero from chronic placental insufficiency. These findings highlight the unique pattern of mortality and morbidity seen in infants with severe early IUGR. PMID- 15116140 TI - Nosocomial infection in the NICU: a medical complication or unavoidable problem? AB - Nosocomial sepsis is a serious problem for neonates who are admitted for intensive care. As it is associated with increases in mortality, morbidity, and prolonged length of hospital stay, both the human and fiscal costs of these infections are high. Although the rate of nosocomial sepsis increases with the degree of both prematurity and low birth weight, no specific lab test has been shown to be very useful in improving our ability to predict who has a "real" blood-stream infection and, therefore, who needs to be treated with a full course of antibiotics. As a result, antibiotic use is double the rate of "proven" sepsis and we are facilitating the growth of resistant organisms in the neonatal intensive care unit. The purpose of this article is to review the topic of nosocomial infections in neonates. PMID- 15116141 TI - Dye-determined amniotic fluid volume and intrapartum/neonatal outcome. AB - OBJECTIVE: To ascertain if a dye-determined amniotic fluid volume was predictive of intrapartum and perinatal outcome. MATERIALS AND METHODS: The low and normal amniotic fluid volumes (< 5th percentile and > or =5th percentile for gestational age) and the raw dye-determined amniotic fluid distributions were correlated with 10 clinical outcome measures in 74 pregnancies. RESULTS: In this observational study, median gestational age at delivery was 36 weeks (range 26 to 41) and 16 deliveries were for fetal distress (14 Cesarean and two forceps). There were no differences between the outcomes of pregnancies with low and normal amniotic fluid volumes for any of the clinical outcomes (variable decelerations influencing delivery, p=0.381; late decelerations, p=0.875; Cesarean births for fetal intolerance of labor, p=0.259; intrauterine growth restriction, p=0.998; or umbilical cord arterial pH< 7.2, p=0.259). Analogous results were obtained when the gestational age-adjusted amniotic fluid volumes were compared directly between the pregnancies with normal and abnormal outcomes. There was no difference between the mean amniotic fluid volumes in those pregnancies with variable decelerations influencing delivery (p=0.287), late decelerations (p=0.555), Cesarean births for fetal intolerance of labor (p=0.310), intrauterine growth restriction (p=0.267) or umbilical cord arterial pH< 7.2, and the pregnancies without these intrapartum events. Reduced variability was more commonly observed in pregnancies with higher amniotic fluid volumes (p=0.038, 771 ml, 95% CI 468 to 1269, compared to those without normal variability 444 ml, 95% CI 374 to 526). CONCLUSIONS: Dye-determined amniotic fluid volume does not appear to be predictive of adverse intrapartum and neonatal outcome. PMID- 15116142 TI - An economic evaluation of the Irbesartan in Diabetic Nephropathy Trial (IDNT) in a UK setting. AB - There are substantial healthcare costs associated with the provision of renal replacement therapy. Patients with diabetes mellitus are the largest and fastest growing group developing end-stage renal disease (ESRD) in the United Kingdom (UK). Treatment leading to a slowing of progression to ESRD in diabetic patients could lead to considerable cost savings. Using treatment-specific probabilities derived from the Irbesartan in Diabetic Nephropathy Trial (IDNT), the cost effectiveness of treating patients with hypertension, type II diabetes and nephropathy with irbesartan, amlodipine or control was calculated using a Markov model. UK-specific ESRD-related data were retrieved from published sources to reflect local management practices, ESRD outcomes and costs. Mean 10-year costs and changes in life expectancy due to ESRD delayed or avoided were calculated. Future costs and clinical benefits were discounted at 6.0 and 1.5% per annum and extensive sensitivity analyses were performed. Delay in the onset of ESRD with irbesartan led to cost savings of pound sterling 5125 and pound sterling 2919/patient and improvements in projected discounted life expectancy of 0.07 and 0.21 years over 10 years vs amlodipine and control, respectively. The costs of treatment of ESRD were the main contributor to the total costs. The cost of trial medications had only a minor impact. These results were robust in a wide range of plausible assumptions. Given that the IDNT efficacy results could be translated to a UK setting, treating patients with hypertension, type II diabetes and overt nephropathy with irbesartan was cost saving over a 10-year period compared to amlodipine and control. PMID- 15116143 TI - Baroreflex sensitivity, heart rate, and blood pressure variability in hypertensive pregnancy disorders. AB - Hypertensive pregnancy disorders are a leading cause of perinatal and maternal morbidity and mortality. Heart rate variability (HRV), blood pressure variability (BPV), and baroreflex sensitivity (BRS) are relevant predictors of cardiovascular risk in humans. The aim of the study was to evaluate whether HRV, BPV, and BRS differ between distinct hypertensive pregnancy disorders. Continuous heart rate and blood pressure recordings were performed in 80 healthy pregnant women as controls (CON), 19 with chronic hypertension (CH), 18 with pregnancy-induced hypertension (PIH), and 44 with pre-eclampsia (PE). The data were assessed by time and frequency domain analysis, nonlinear dynamics, and BRS. BPV is markedly altered in all three groups with hypertensive disorders compared to healthy pregnancies, whereby changes were most pronounced in PE patients. Interestingly, this increase in PE patients did not lead to elevated spontaneous baroreflex events, while BPV changes in both the other hypertensive groups were paralleled by alterations in baroreflex parameters. The HRV is unaltered in CH and PE but significantly impaired in PIH. We conclude that parameters of the HRV, BPV, and BRS differ between various hypertensive pregnancy disorders. Thus, distinct clinical manifestations of hypertension in pregnancy have different pathophysiological, regulatory, and compensatory mechanisms. PMID- 15116144 TI - Prevalence and correlates of orthostatic hypotension in middle-aged men and women in Korea: the Korean Health and Genome Study. AB - To examine the prevalence and correlates of orthostatic hypotension (OH) in middle-aged adults enrolled in the Korean Health and Genome Study. Participants were 8908 individuals aged 40-69 years. Supine blood pressure (BP) was measured three times at 30-s intervals after at least 5 min of rest in the supine position and single standing BP was measured at 0 and 2 min after standing, respectively. OH was defined as a reduction in systolic BP or diastolic BP > or = 20 and 10 mmHg, respectively. The prevalence of OH at 0 and 2 min after standing was 12.3 and 2.9%, respectively. At 0 min of standing, OH frequency increased significantly with age from 6.4% in those aged 40-44 years to 23.1% in those aged 65-69 (P < 0.001). After adjustment for age and other characteristics, hypertension was associated with a 1.7-fold excess in the odds of OH in men and a 1.6-fold excess in women (P < 0.001). In contrast, an increase in body mass index (BMI) on the order of 5 kg/m2 was associated with a 20-30% reduction in the odds of OH (P < 0.001). Diabetes in women was also associated with a 1.4-fold excess in the odds of OH (P < 0.05). An increase in triglyceride by 136 mg/dl in men was associated with an increase in the odds of OH (P < 0.05). In conclusion, the prevalence and correlates of OH other than diabetes and triglycerides were notably similar in men and women. While the association between hypertension and OH has been observed elsewhere, low BMI in Korean adults with OH may be an important marker for subclinical morbidity or coexisting risk factors that need to be identified. PMID- 15116145 TI - Strategies for improvement of awareness, treatment and control of hypertension: results of a panel discussion. AB - High blood pressure (BP) is a major risk factor for coronary heart disease, heart failure, stroke, chronic kidney disease, end stage renal disease, and a variety of other clinically important outcomes. Results from the surveys described in this issue and elsewhere underscore a common finding that hypertension is both highly prevalent and insufficiently treated and controlled. Recognizing the differences in sampling and survey measurement techniques, the reported prevalence of hypertension (SBP/DBP >/=140/90 mmHg or treatment with antihypertensive medication) in adults exceeded 25% in all of the surveys reported in this issue. In Latvia, the prevalence of hypertension for 25-64-year old adults in the general population was 46.1%. Control of hypertension with medication to an SBP/DBP <140/90 mmHg in the general population ranged from as low as 12% to a high of only 29%. Data from other parts of the world provide an equally distressing picture of what is (not) being accomplished in treatment and control of hypertension at the level of the general population. These data provide testimony to an urgent need for greater attention to the treatment and control of hypertension in populations around the world. This was the basis for a panel discussion at the International Society of Hypertension satellite conference The Epidemiology of Hypertension-Regional Differences in Treatment and Control. Panel participants included Drs P Whelton, S Sonkodi, DG Beevers, JG Fodor, H Elliot, R Cifkova, A Nissinen, A Javor, and there was active participation of other symposium attendees. The following summarizes key elements of the discussion and recommendations of the panel. PMID- 15116146 TI - [The "rehabilitation" of the National Health Service]. PMID- 15116147 TI - [Microbiological monitoring activities: a report in a food-crockery-cleaning factory]. AB - The purpose of this report is to describe the checks, the methods and the results obtained in a crockery-cleaning and polishing factory. Investigating the most important biological index for food contamination (e.g. T.B.C., Yeast, Mould), in order to determine the presence/absence of biological agent and to obtain good results in terms of hygiene and health. PMID- 15116148 TI - [Effectiveness of the maintenance operations on the air conditioning systems of a university building in relation to the microbiological quality of the air indoor]. AB - The microbiological quality of the air indoor is influenced from various factors and one of the most important is represented from the maintenance of the conditioning systems. In this study it has been estimated the effectiveness of an intervention of cleaning and maintenance on the systems of conditioning of an university building executing sampling before and after such intervention. The two results were confronted and it is observed as the maintenance of the air conditioners has influenced on the quality of the air indoor. PMID- 15116149 TI - [Adherence to the screening program for HBV infection in pregnant women and application of the immunoprophylaxis protocol in babies born to HBV carrier women]. AB - The aim of the study was to evaluate the frequency of the screening for HBV infection in pregnant women and the application of immunoprophylaxis in newborns to HBV carrier women. The study, carried out in 2001 for 2 months, involved 1596 pregnant women consecutively recruited in public and private hospitals of the Sardinia. Information has been collected by a questionnaire: 90.5% of the women underwent HBV screening during pregnancy and 2.6% of them were found positive to HBsAg test. Among the newborns to HBsAg carrier mothers, 89.2% received the immunoprophylaxis protocol (specific immunoglobulin and the first dose of vaccine within 24 hours from birth). Two variables resulted statistically associated with the lack of adherence to HBV screening: the original family size of women (more than 4 members in the household) and the delivery in a private hospital. These findings point out a satisfactory adherence to HBV screening during pregnancy and the application of immunisation protocol in most of the newborns to HBsAg carrier mothers. However, the increase of information programs turned to the population is required to reach the total application of the prevention tools available in Italian public health. PMID- 15116150 TI - [Tetanus in Italy and in the Campania region: retrospective analysis. The importance of vaccinations]. AB - The authors intend to draw the attention on the issue of tetanus infections in adult patients whose history is often unknown or incomplete. For these people, current regulations (Presidential Decree 7th Nov 2001 no. 464) provides for more extended vaccination coverage. The input data were drawn by different sources (ISS; MS; National Statistics Institute; Campania Epidemiological Observatory) and allowed for the tetanus infections having occurred in Italy and Campania in the last ten years. PMID- 15116151 TI - [Waiting lists: case management and an alternative reimbursement paradigm]. AB - Long-waiting lists to undergo preliminary medical examinations and treatments are a tough problem for western countries. Many actions and programs have been taken to offer more health services, which initially shortened the waiting lists. However, after this initial alleviation, the later demand for further diagnostic examinations inevitably creates subsequent increases in diagnostic service waiting times. In this article the authors suggest some solutions to manage this problem: creating organized procedures for case management, homogeneous waiting list reporting systems, outpatient procedures for white triage and diagnostic services opening overtime during non-working days. An alternative form of reimbursement is also suggested. PMID- 15116152 TI - [The Romanov Report on the future of Canadian health care]. PMID- 15116153 TI - [Policies and norms on obligatory vaccination]. PMID- 15116154 TI - Acute myocardial infarction in the elderly. Is primary coronary angioplasty the treatment of choice? In-hospital follow-up results. AB - BACKGROUND: Acute myocardial infarction (AMI) is one of the main causes of death in the elderly, however, the optimal therapy of AMI in this age-group has not yet been established. AIM: To compare the early outcome of patients with AMI aged > or =75 years who underwent primary percutaneous coronary interventions (PCI) or were treated conservatively. METHODS: In-hospital outcome of 180 patients aged > or =75 years, hospitalised due to AMI, was retrospectively analysed. Ninety two consecutive patients, admitted with AMI between May 2001 and October 2002, underwent primary PCI whereas 88 patients, treated for AMI between 1993 and 2002, received standard pharmacological therapy without thrombolysis. CONCLUSIONS: Primary PCI in the elderly with AMI is significantly more effective than conservative therapy, except with patients with cardiogenic shock in whom mortality is similar regardless of the therapy used. PMID- 15116155 TI - Coronary flow reserve in hypertrophic obstructive cardiomyopathy assessed by intracoronary Doppler catheter. AB - BACKGROUND: In spite of progress in diagnosis and treatment, hypertrophic obstructive cardiomyopathy (HOCM) remains a serious medical problem. Among many issues, the pathophysiology of the coronary circulation in HOCM has not yet been fully examined. AIM: To assess coronary flow reserve in HOCM. METHODS: The study group consisted of 15 patients (6 males, 9 females, mean age 51+/-15 years) with typical echocardiographic signs of HOCM and without stenosis of the coronary arteries. Using an intracoronary Doppler catheter, the average peak velocity and the absolute coronary flow reserve were determined in the proximal, medial and distal parts of the left anterior descending (LAD) and the circumflex coronary artery (Cx) following administration of papaverine, substance P, and during pacing. The coronary square plane was calculated angiographically after substance P injection. The retrograde coronary flow and the relationship between the increase of the coronary square plane and the coronary flow reserve were also examined. RESULTS: Under pharmacological stimulation, higher values of the average peak velocity were observed compared to pacing. A retrograde flow was observed in 8 of 10 patients in the LAD and in 3 of 8 patients in the Cx. The coronary flow reserve was higher under pharmacological stress than during pacing. No relationship was found between the increase of the coronary square plane and the coronary flow reserve. CONCLUSIONS: No decrease in the coronary flow reserve was observed in our patients with HOCM which, however, does not exclude the possibility of ischaemia based on subordinate vessels and microcirculation changes. In the majority of patients a retrograde flow was detected. PMID- 15116156 TI - Effects of coronary artery by-pass grafting on haemostatic factors. AB - BACKGROUND: Atherogenesis is triggered by functional or structural endothelial injury which leads to humoral and secretion alterations. The haemostatic system plays a significant role in the development of atherosclerosis and its complications, especially acute coronary syndromes. AIM: To assess the effects of coronary artery by-pass grafting (CABG) on some haemostatic factors in patients with stable coronary artery disease (CAD). METHODS: The study group consisted of 45 patients (11 females, 34 males, mean age 60.4+/-9.4 years, range 35-75 years). Endothelial function marker - von Willebrand factor (vWF), and thrombin generation parameters - thrombin-antithrombin complexes (TAT) and prothrombin fragments - F1+2 were measured before as well as 3 days, 7 days and 3 months after CABG. The control group consisted of 33 healthy volunteers (17 females, 16 males, mean age 48.8+/-17.2 years). RESULTS: Serum concentration of vWF as well as TAT and F1+2 levels were significantly higher in the CABG group than in controls. Compared with baseline values, the vWF levels were significantly higher 3 and 7 days after CABG. CONCLUSIONS: There is a hypercoagulability state in patients with CAD who undergo CABG. The use of extracorporeal circulation causes a significant increase in the vWF level which suggests a marked endothelial injury caused by CABG procedure. PMID- 15116158 TI - The biphasic course of changes of left ventricular outflow gradient after alcohol septal ablation for hypertrophic obstructive cardiomyopathy. AB - BACKGROUND AND AIM: Alcohol septal ablation (PTSMA) decreases left ventricular outflow gradient (LVOG) and relieves symptoms in patients with hypertrophic obstructive cardiomyopathy (HOCM). The time course of early changes of LVOG has not been clearly determined up to now. METHODS: Doppler echocardiography was used to determine the maximal LVOG. Thirty-nine consecutive patients (24 women; age 55+/-14) were examined at baseline, immediately after the procedure, and 3-5 days, 3 weeks and 3 months thereafter. RESULTS: The baseline LVOG decreased immediately after PTSMA from 73+/-49 mmHg to 13+/-16 mmHg (p<0.01). During the hospital stay (postprocedural period) LVOG increased from 13+/-16 to 37+/-35 mmHg (p<0.01). All the patients were discharged 5-10 days after the procedure. At three-week examination LVOG decreased from 37+/-35 to 25+/-12 mmHg (p<0.01). Three-month survival was 97%. One patient died suddenly one month after PTSMA. At three-month examination LVOG decreased from 25+/-12 to 17+/-14 mmHg (NS). All the patients reported an improvement in symptoms at follow-up. CONCLUSIONS: The immediate decrease of LVOG after PTSMA procedure caused by myocardial necrosis and stunning, was followed by a significant LVOG increase during the early postprocedural period and continuous LVOG decrease at the short-term follow-up. PMID- 15116157 TI - Effects of cardiopulmonary bypass on the expression of adhesive molecules in children undergoing cardiac surgery due to congenital heart disorders. AB - BACKGROUND: Infants and small children who undergo cardiac surgery due to congenital heart disorders, are at risk of developing inflammatory complications and multi-organ failure. AIM: To assess the changes in the expression of adhesive molecules on neutrophils and in peripheral blood plasma in children undergoing cardiac surgery with or without the use of cardiopulmonary bypass (CPB). METHODS: In 18 children who underwent surgery with the use of CPB and in 7 children who underwent surgery without CPB, the expression of CD11b, CD11c and CD62L molecules on neutrophils and soluble adhesive molecules - L-selectin and ICAM-1, was assessed before, during and after cardiac surgery. RESULTS: A significant increase in the expression of CD11b on neutrophils at the time of the initiation of CPB was observed. The most pronounced decrease in the expression of CD11c was detected at the end of surgical procedure. There was also a negative correlation between L-selectin concentration measured at the end of CPB and CPB duration as well as between ICAM-1 concentration and duration of hypothermia. CONCLUSIONS: Cardiac surgery influences the expression of adhesive molecules on neutrophils and in blood plasma. These changes are more pronounced in children who undergo CPB, and depend on CPB duration as well as the use of hypothermia. PMID- 15116159 TI - [Pericarditis as a consequence and first symptom of microcellular lung carcinoma- case report]. AB - A case of a patient suffering from Streptococcal suppurative pericarditis with cardiac tamponade and acute renal failure as a first symptom of disseminated lung cancer is presented. The examination of pericardial fluid only confirmed Streptococcal infection as a cause of exudative pericarditis. The neoplasmatic cells were not found in the examined fluid. Next, chest CT and bronchoscopy were performed due to the lack of clinical improvement, recurrence of severe pericardial effusion and enlargement of the upper mediastinum found on chest radiogram. CT and bronchoscopy allowed to establish the final diagnosis - disseminated lung cancer. PMID- 15116160 TI - [Infective endocarditis caused by Erysipelothrix rhusiopathiae--case report]. AB - Infective endocarditis (IE) is an infectious disease, which leads to death when is untreated. In most cases IE is caused by typical bacteria. IE caused by atypical bacteria is rare. In this paper, we describe a female patient with IE caused by Erysipelothrix rhusiopathiae (ER). Due to inflammation and leaflets' damage she underwent double-valve implantation (aortic and mitral ones). A long term antibiotic therapy was given with good outcome. We also describe the organism, types of human diseases caused by ER and treatment options. PMID- 15116161 TI - [Pheochromocytoma mimicking acute myocarditis]. AB - Pheochromocytoma, a relatively rare (0.1-0.8% of hypertensives), catecholamine secreting tumor, is almost always lethal unless recognised and appropriately treated. Histological changes in myocardium can be often found in patients with hypertension crisis due to pheochromocytoma, however, only a few patients with clinical manifestations of acute myocarditis were described. We report a 18-year old man hospitalised with symptoms of acute myocarditis. Precise clinical assessment allowed to diagnose pheochromocytoma. PMID- 15116162 TI - [Calcium cycling in myocytes of normal and failing heart. New perspectives for therapy]. PMID- 15116164 TI - [Cough cardiopulmonary resuscitation]. PMID- 15116163 TI - [Natriuretic peptides in atrial fibrillation. State of art and new concepts]. PMID- 15116166 TI - [Cardiologic health care in Poland--report from cardiology consultants in the year 2003]. PMID- 15116168 TI - [Perforated aneurysm of posterior mitral leaflet]. PMID- 15116169 TI - [Electrocardiogram of the month]. PMID- 15116170 TI - [Role of electrophysiologic properties of the A-V node and His-Purkinje system in triggering inappropriate ICD discharges in a patient with arrhythmogenic right ventricular dysplasia. Effective treatment with RF ablation]. PMID- 15116171 TI - [Angiogram of the month]. PMID- 15116173 TI - [Letters to the editor]. PMID- 15116172 TI - [Usefulness of baroreflex sensitivity in clinical cardiology]. PMID- 15116175 TI - Enantioselective chemo- and bio-catalysis in ionic liquids. AB - Recent developments in the enantioselective chemo- and bio-catalysis in ionic liquids are reviewed. In many cases, the use of ionic liquids provides many advantages over reactions in conventional organic solvents in terms of activity, enantioselectivity, stability and the reusability of the solvent-catalyst systems. PMID- 15116177 TI - A novel ligand-unsupported 3D framework polymer of trimeric copper(I) and its NLO property. AB - A novel 3-D coordination polymer with trimeric copper (I) unit, [Cu(3)(CN)(IN)(2)](n)(IN = isonicotinate), was hydrothermally synthesized by the reaction of Cu(NO(3))(2).3H(2)O with isonicotinic and terephthalic acids. The structure was characterized to be a twofold interpenetrated 3-D coordination network polymer with two-coordinated copper(I). It has a powder SHG efficiency about that of KDP. PMID- 15116176 TI - Synthesis of peptide-nanotube platinum-nanoparticle composites. AB - Nanotubes prepared by the self-assembly of D-Phe-D-Phe molecules are investigated by electron microscopy and Monte Carlo simulations; the nanotubes appear to be porous and are capable of forming novel peptide-nanotube platinum-nanoparticle composites. PMID- 15116178 TI - Ferromagnetic NiII-GdIII interactions in complexes with NiGd, NiGdNi, and NiGdGdNi cores supported by tripodal ligands. AB - Dinuclear [(NiL)Gd(hfac)(2)(EtOH)](H(3)L = 1,1,1-tris(N salicylideneaminomethyl)ethane, Hhfac = hexafluoroacetylacetone), trinuclear [(NiL)(2)Gd(NO(3))], and tetranuclear [(NiL)Gd(CH(3)CO(2))(2)(MeOH)](2) complexes, were prepared by treating [Ni(HL)] with [Gd(hfac)(3)(H(2)O)(2)], Gd(NO(3))(3).6H(2)O, and Gd(CH(3)CO(2))(3).4H(2)O, respectively, in the presence of Et(3)N. All the complexes show that ferromagnetic interactions occur between the Ni(II) and Gd(III) ions. PMID- 15116179 TI - A novel fluoride sensor based on fluorescence enhancement. AB - A novel halide sensor, which yields greater fluorescence upon binding to fluoride, has been synthesized and characterized. PMID- 15116180 TI - Solid-state caging of 1,10-phenanthroline pi-pi stacked dimers by calix[4]arene dihydroxyphosphonic acid. AB - The calix[4]arene dihydroxyphosphonic acid-1,10-phenanthroline complex shows caging of the guest molecules as a pi-pi stacked dimer in a cavity formed by intermolecular hydrogen bonds and aromatic walls formed by the calixarene. PMID- 15116181 TI - Reactions of N-heterocyclic carbenes (NHCs) with one-electron oxidants: possible formation of a carbene cation radical. AB - One-electron oxidation of N-heterocyclic carbenes (NHCs) has been carried out using oxidising agents such as tetracyanoethylene (TCNE) and ferrocenium [Cp(2)Fe](+); the formation of carbene radical cations is postulated. PMID- 15116182 TI - Self-assembly of a novel pentanuclear centred-tetrahedral silver species. AB - The reaction of 3,6-di(2-pyridyl)-4-trimethylsilylpyridazine with silver(I) tetrafluoroborate gives the pentanuclear complex cation [Ag(5)L(4)](5+), in which the silver centres are arranged as a centred tetrahedron. PMID- 15116183 TI - Helical and network coordination polymers based on a novel C2-symmetric ligand : SHG enhancement through specific metal coordination. AB - Cu(I) and Ag(I) coordination polymers of an axially chiral "push-pull" ligand possess respectively 2-D network and helical structures and the coordination mode strongly influences the solid state SHG reduction/enhancement with respect to the free ligand. PMID- 15116185 TI - A surprising steric effect on a tandem cycloaddition/ring-opening reaction: rapid syntheses of difluorinated analogues of (hydroxymethyl)conduritols. AB - Difluorinated analogues of (hydroxymethyl)conduritols can be synthesised from selected furans and a difluorinated dienophile in two reaction steps. PMID- 15116184 TI - Octaethylporphyrin and expanded porphyrin complexes containing coordinated BF2 groups. AB - In contrast to octaethylporphyrin, which forms a very labile bis-BF(2) complex, treatment of the hexa- and octapyrrolic expanded porphyrins amethyrin and [32]octaphyrin with BF(3).Et(2) under standard reaction and work-up conditions gives rise to stable, non-labile mono- and bis-BF(2) complexes; these were readily characterised by, inter alia, X-ray diffraction analyses. PMID- 15116186 TI - DNA-selective hybridization and dual strand invasion of short double-stranded DNA using pyren-1-ylcarbonyl-functionalized 4'-C-piperazinomethyl-DNA. AB - Incorporation of a novel pyren-1-ylcarbonyl-functionalized 4'-C-piperazinomethyl DNA monomer into oligodeoxynucleotides leads to increased thermal stability of duplexes with DNA complements but reduced thermal stability of duplexes with RNA complements. This DNA-selective hybridization is explored for recognition of double-stranded DNA by a novel dual strand invasion approach. PMID- 15116187 TI - Exploiting phenyl embraces and pi-stacking in the assembly of arrays of tetraphenylphosphonium p-sulfonatocalix[4]arene. AB - Phenyl embraces involving tetraphenylphosphonium cations feature in complexes of p-sulfonatocalix[4]arene where a phenyl ring of a cation is included in the cavity of the calixarene. The overall structures are based on pseudo-polymorphic supramolecular arrays and their formation is templated or induced by lanthanide ions. PMID- 15116188 TI - Hexakis(isopropylthio)-1,5-hexadien-3-yne and its fluorescent Ag(I) coordination polymers: assembly of helicates with thioether sites. AB - A new thioether-rich ligand with a conjugated dienyne backbone and its fluorescent Ag(I) coordination networks have been synthesized and characterized by single crystal X-ray diffraction studies, which reveal that the supramolecular architectures of the networks contain assembled helicates with thioether sites. PMID- 15116190 TI - Conversion of alcohols into N-alkyl anilines via an indirect aza-Wittig reaction. AB - Iridium catalysed oxidation of alcohols provides the aldehydes required for in situ aza-Wittig reactions and the so-formed imines are reduced to amines under the reaction conditions. PMID- 15116189 TI - Carbon monoxide solubility in ionic liquids: determination, prediction and relevance to hydroformylation. AB - The solubility of carbon monoxide in 37 ionic liquids and in some organic solvents has been determined using high-pressure (13)C NMR spectroscopy; a method for predicting the CO solubility is demonstrated, and it was shown that the rate of the hydroformylation of 5-hexen-2-one does not correlate with the CO solubility, as expected from the determined relative solubility of CO compared to H(2). PMID- 15116191 TI - Ultrathin hollow nanoshells of manganese oxide. AB - Novel hollow nanoshells of Mn(2)O(3) with controllable ultrathin shell thickness have been fabricated through layer-by-layer assembly of exfoliated MnO(2) nanosheets and polyelectrolytes on polymer bead templates, followed by removal of the polymer cores via calcination. PMID- 15116192 TI - Sulfonium ylide epoxidation reactions: methylene transfer. AB - Using a D-mannitol derived chiral sulfide, terminal epoxides are formed in up to 76% ee; the first example of double asymmetric induction in a sulfonium methylide epoxidation is reported and an improved method of generating sulfonium ylides is detailed. PMID- 15116193 TI - Straightforward synthesis of gem-phosphonate-phosphate containing compounds via one-pot reaction of thioesters with diethyl phosphite. AB - A convenient straightforward, three step, one-pot reaction of diethyl phosphite with thioesters in the presence of catalytic amounts of potassium tert-butoxide in toluene or DMF as solvent provided various phosphonate-phosphate containing compounds in excellent yields. PMID- 15116194 TI - An iterative approach to novel polyamines via nucleophilic ring-opening of aziridinium ions with beta-amino alcohols. AB - An iterative procedure for the synthesis of a novel class of synthetic polyamines has been developed, utilising the regioselective ring-opening of aziridinium ion intermediates; facile N-allyl deprotection of intermediate polyamines allows the rapid construction of high molecular weight, stereochemically defined compounds in a convergent manner. PMID- 15116195 TI - Consideration of spin states in determining the structure and decomposition of the transition metal pentazoles FeClN5, Fe(N5)2, Fe(H2O)4ClN5, and Fe(NH3)4ClN5. AB - Optimised geometries and energies have been calculated for the unidentate, bidentate, and ferrocene-like structures of Fe(II) pentazoles of the form MN(5) and M'(N(5))(2), (M = FeCl, M'= Fe) using the UB3LYP DFT method with basis sets up to 6-311+G(3df) in order to study the effect of an unfilled d shell on the stability of the singlet, triplet, and quintet states; the ground state is a quintet and the quintet has a lower barrier to decomposition than the singlet. PMID- 15116196 TI - Use of an ionic liquid in a two-phase system to improve an alcohol dehydrogenase catalysed reduction. AB - Due to favourable partition coefficients the highly enantioselective reduction of 2-octanone, catalysed by an alcohol dehydrogenase from Lactobacillus brevis, is faster in a biphasic system containing buffer and the ionic liquid [BMIM][(CF(3)SO(2))(2)N] compared to the reduction in a biphasic system containing buffer and methyl tert-butyl ether. PMID- 15116197 TI - A new entry to Amaryllidaceae alkaloids from carbohydrates: total synthesis of (+)-vittatine. AB - The stereoselective and chiral synthesis of the Amaryllidaceae alkaloid, (+) vittatine 1, is described; the quaternary carbon in 1 was generated by Claisen rearrangement of a cyclohexenol derived from D-glucose by way of a Ferrier's carbocyclisation reaction and a hexahydroindole skeleton was effectively constructed by intramolecular aminomercuration-demercuration, followed by Chugaev reaction. PMID- 15116198 TI - Model systems for flavoenzyme activity: an electrochemically tuneable model of roseoflavin. AB - The electrochemically tuneable hydrogen bonding interactions between roseoflavin analogue and apoenzyme mimic are described. PMID- 15116199 TI - Porphyrin ring contraction: a one-pot reaction leading to divalent corroles. AB - Reaction of nickel meso-tetraarylporphyrins with benzoic anhydride and tin tetrachloride, followed by air oxidation under basic conditions, results in ring contraction, yielding divalent corroles. PMID- 15116200 TI - Organization of branched rod-coil molecules into a 3-D tetragonally perforated lamellar mesophase. AB - Tetramerization of coil-rod-coil ABC triblock copolymers to a tetrabranched molecule induces an unusual 3-D tetragonally perforated layered liquid crystalline phase as an intermediate structure between 1-D lamellar and 2-D hexagonal columnar phases. PMID- 15116201 TI - DPA-substituted coumarins as chemosensors for zinc(II): modulation of the chemosensory characteristics by variation of the position of the chelate on the coumarin. AB - The sensory capabilities of two novel di(2-picolyl)amine (DPA)-substituted coumarins are described and it is shown that the variation of the point of attachment of the DPA group to the coumarin framework controls their sensing behavior: the 4-substituted system is a CHEF-type sensor that shows a significant increase in fluorescence intensity upon Zn(2+) binding, whereas the 3-substituted system is a ratiometric sensor. PMID- 15116202 TI - Supported choline hydroxide (ionic liquid) as heterogeneous catalyst for aldol condensation reactions. AB - Choline hydroxide was used as a basic catalyst for aldol condensation reactions to produce new C-C bonds between several ketones and aldehydes. Choline supported on MgO exhibits higher TOF values than other well known basic catalysts in these reactions. PMID- 15116203 TI - Involvement of heteroatoms in charge transfer of Keggin-type heteropolyanion. AB - Spectroscopic measurements of an electrochemically deposited film from a Keggin type heteropolyanion solution indicate that electron transfer takes place not only on polyatoms but also on heteroatoms of silicon. PMID- 15116204 TI - Two mixed-valence copper(I,II) imidazolate coordination polymers: metal-valence tuning approach for new topological structures. AB - A new metal-valence tuning synthetic approach has been utilized to generate two new mixed-valence Cu(I,II) coordination polymers Cu(2)(im)(3) and Cu(3)(im)(4)(Him = imidazole), which are an unprecedented uninodal 4-connected 4.8(5) topological net and a 4-connected (4,4) net, respectively. PMID- 15116205 TI - A new tetrameric CuII cluster with square topology exhibiting ferro- and antiferromagnetic magnetic pathways : which is which? AB - [Cu(4)L(2)(bpy)(4)(H(2)O)(3)](ClO(4))(4).2.5H(2)O, 1, a new tetranuclear Cu(II) cluster showing square planar geometry, formed with aspartate bridging ligand (L) has been synthesized. The global magnetic coupling is ferromagnetic but theoretical DFT/B3LYP calculations are necessary to assign which Cu-L-Cu side is ferro or antiferromagnetically coupled. PMID- 15116206 TI - Synthesis of gold nanoplates by aspartate reduction of gold chloride. AB - Single crystal nanoplates with thickness less than 30 nm, characterized by hexagonal and truncated triangular shapes bounded mainly by [111] facets, were obtained in large quantities by aspartate reduction of gold chloride. PMID- 15116207 TI - Synthesis of silver nanotubes by electroless deposition in porous anodic aluminium oxide templates. AB - An electroless deposition method has been employed for the synthesis of silver nanotubes using porous anodic aluminium oxide as templates, by which high-yield silver nanotubes with length over ten microns have been synthesized. PMID- 15116208 TI - Baylis-Hillman adducts in rhodium-catalyzed 1,4-additions: unusual reactivity. AB - In the presence of a rhodium catalyst, unactivated Baylis-Hillman adducts reacted with arylboronic acids to afford trisubstituted alkenes with good yields. This highly efficient reaction (aerobic conditions, low temperature) is believed to proceeds via an unexpected mechanism involving 1,4-addition/beta-hydroxy elimination steps and not pi-allyl type rhodium intermediates. PMID- 15116209 TI - Two-dimensional molecular ordering of Os(II) complexes in organo-clay hybrid ultrathin films. AB - An ordered structure of [OsL(3)](2+)(L = 2, 2'-bipyridine or 1, 10 phenanthroline) layer in an ultrathin film composed of the complex cations, clay nanosheets and alkylammonium cations is reported on the basis of its in-plane X ray diffraction and polarized electronic spectral data. PMID- 15116210 TI - Towards configurationally stable bisindolylmaleimide cyclophanes: potential tools for investigating protein kinase function. AB - The effect of macrocycle size and substitution on the configurational stability of some bisindolylmaleimide cyclophanes was determined. PMID- 15116211 TI - Synthesis and noncovalent polymerization of self-complementary hydrogen-bonding supramolecular synthons: N,N'-disubstituted 4,6-diamino-pyrimidin-2(1H)-ones. AB - A series of self-complementary N,N'-disubstituted 4,6-diaminopyrimidin-2(1H)-ones self-assemble in organic solvents to form a robust and linear supramolecular polymer network via DDA.AAD hydrogen-bonding. PMID- 15116212 TI - Palladium(II) chloride catalyzed selective acetylation of alcohols with vinyl acetate. AB - PdCl(2) can catalyze the acetylation of primary and secondary alcohols with vinyl acetate. The reaction is selective and mild with high yields. Tertiary alcohols, phenols and amines are unaffected under these reaction conditions. PMID- 15116213 TI - Ligand effects on the electrochemical and spectroscopic behaviors of methano[60]fullerene derivatives. AB - Effects of the ligand chemical structures on the electronic energy properties of two methano[60]fullerene derivatives were investigated voltammetrically and spectroscopically. PMID- 15116214 TI - Analysis of liposomal membrane composition using Raman tweezers. AB - We have developed a methodology for the analysis of liposomal membranes and their contents using near-IR Raman spectroscopy on liposomes held in an optical trap. We were able to detect a variety of membrane components including lipids, cholesterol, and small molecule solutes such as ethanol, DMSO and hexafluoroisopropanol. The methodology is able to distinguish between solutes that equilibrate across the liposomal membrane from those that partition selectively into the lipid bilayer. PMID- 15116215 TI - Stabilization of enhanced chirality from pyrene-containing l-glutamide lipid in methyl methacrylate by photo-induced polymerization. AB - Enhanced CD spectra based on chirally-oriented structures of pyrene-containing L glutamic acid-derived lipid were observed in polymerizable monomers and CD strength was maintained after photo-induced polymerization of the monomer solvents. PMID- 15116216 TI - Palladium-catalysed coupling reaction of allenic alcohols with aryl- and alkenylboronic acids. AB - The direct coupling of aryl- and alkenylboronic acids with allenic alcohols has been achieved using a palladium catalyst to yield various substituted dienes and trienes in high yields. PMID- 15116217 TI - Soluble mu-Fi bridged niobium clusters: synthesis and crystal structures of (Et4N)6[Nb6Fi6Bri6(NCS)a6]Br2 and Cs1.6K2.4[Nb6Fi6Ii6(NCS)a6]. AB - Two new anions [Nb(6)F(i)(6)X(i)(6)(NCS)(a)(6)](4-)(X = Br, I) based on octahedral niobium clusters with edge-bridging F ligands have been prepared by reaction of Cs(3)Nb(6)F(6)Br(12) and Cs(4)Nb(6)F(8.5)I(9.5) with aqueous solution of KSCN. The anions were isolated as (Et(4)N)(6)[Nb(6)F(6)Br(6)(NCS)(6)]Br(2) (1)and Cs(1.6)K(2.4)[Nb(6)F(6)I(6)(NCS)(6)] (2) salts. PMID- 15116218 TI - Double asymmetric induction as a mechanistic probe: conjugate addition for the asymmetric synthesis of a pseudotripeptide. AB - Double asymmetric induction as a mechanistic probe indicates that, for the conjugate addition of (R)- and (S)-lithium N-benzyl-N--alpha-methylbenzylamide to (S)-3'-phenylprop-2'-enoyl-4-benzyloxazolidinone, the reactive conformation of the N-acyl oxazolidinone is the anti-s-cis form, facilitating the asymmetric synthesis of a pseudotripeptide. PMID- 15116219 TI - Novel cobalt-free oxygen permeable membrane. AB - A series of cobalt-free and low cost perovskite oxygen permeable membranes based on BaCe(x)Fe(1-x)O(3-delta)(BCF) oxides was successfully synthesized and the membrane showed both high oxygen permeability and high stability under reductive atmosphere, which will be most suitable for constructing a membrane reactor for selective oxidation of light hydrocarbons to syngas or high value corresponding oxygenates. PMID- 15116220 TI - First example of the chemical, oxidative cleavage of the C-P bond in aminophosphonate chemistry. The oxidation of 1-amino-1-(3,4 dihydroxyphenyl)methylphosphonic acid by NaIO4. AB - Oxidative, chemical cleavage of the C-P bond in 1-amino-1-(3,4 dihydroxyphenyl)methylphosphonic acid upon the action of NaIO(4) have been the subject of the NMR, EPR and UV-Vis investigations in acidic and basic conditions. PMID- 15116221 TI - Desymmetrization of meso-dienyne by asymmetric Pauson-Khand type reaction catalysts. AB - Desymmetrization of the meso dienynes, such as propargyl 1-vinylallyl N tosylamides (1a-c) and propargyl 1-vinylallyl ethers (1d-e), by asymmetric Pauson Khand type reaction catalysts was studied. The corresponding vinyl substituted bicyclic pentenones (2 and 3) were obtained with high diastereoselectivity and enantioselectivity. PMID- 15116222 TI - Novel fluorescent biosensor for pathogenic toxins using cyclic polypeptide conjugates. AB - This work describes a two-step conjugate synthesis of a new fluorescent analog of microcystin-LR and its subsequent utilization for the development of an optical biosensor for cyanobacteria toxins. The biosensor concept is based on the competitive binding between the native microcystin and its fluorescent analog at immobilized alkaline phosphatase enzymes. PMID- 15116223 TI - Synthesis of branched poly(methyl methacrylate)s via controlled/living polymerisations exploiting ethylene glycol dimethacrylate as branching agent. AB - With appropriate choice of reaction composition and conditions, copolymerisation of methyl methacrylate and ethylene glycol dimethacrylate using Cu-based ATRP or GTP methodologies yields soluble branched polymers in facile one-pot reactions. PMID- 15116224 TI - Mechanism of the catalytic oxidation of hydrocarbons by N-hydroxyphthalimide: a theoretical study. AB - The mechanism of the recently proposed catalytic oxidation of hydrocarbons by oxygen in the presence of N-hydroxyphthalimide (NHPI) was established by quantum chemical calculations, consistent with experiments. PMID- 15116225 TI - 7Li,31P Shift correlation. Application to the structural assignment of benzyllithium complexes of N-methyl-N-benzylphosphinamide. AB - A correlation experiment between (7)Li and (31)P nuclei through scalar coupling is described for the first time. The utility of the method is demonstrated by identifying the species formed in the benzylic lithiation of N-benzyl-N methyldiphenylphosphinamide in Et(2)O solution. PMID- 15116226 TI - Synthesis and resolution of the planar chirality of ester-functionalised phospharuthenocenes. AB - Menthyl phospholide ester anions provide an operationally simple and high yielding entry to the first planar chiral enantiopure phospharuthenocene derivatives. PMID- 15116227 TI - Determination of arsenic species: a critical review of methods and applications, 2000-2003. AB - We review recent research in the field of arsenic speciation analysis with the emphasis on significant advances, novel applications and current uncertainties. PMID- 15116228 TI - Determination of lower sub ppt levels of environmental analytes using high powered concentration system and high-performance liquid chromatography with fluorescence detection. AB - The proposed method was successful in the determination and separation of lower sub ppt levels of polycyclic aromatic hydrocarbons (PAHs). In this study, a new phase separation phenomenon (i.e., homogeneous liquid-liquid extraction) was developed that rapidly and simply separated an immiscible liquid phase from water miscible organic solvents. One liter of sample solution was preconcentrated by a solid phase extraction method, then the obtained eluate (5 mL) was further preconcentrated to just 20 microL by this homogeneous liquid-liquid extraction. The 20 [micro sign]L sedimented phase was directly injected into the high performance liquid chromatography with fluorescence detection (FL-HPLC). The entire preconcentration factor was 50,000-fold. Six kinds of PAHs were determined in the range of 3.0 x 10(-18) approximately 4.5 x 10(-11) mol L(-1). These chemicals were also satisfactorily separated. PMID- 15116229 TI - Application of selected reaction monitoring tandem mass spectrometry to the quantitative determination of an arsenic-containing nucleoside in a crude biological extract. AB - HPLC electrospray selected reaction monitoring tandem mass spectrometry was used for the determination of a key arsenic metabolite, 5'-dimethylarsinoyladenosine, at trace levels (ca. 20 microg As l(-1)) in a crude extract of kidney from the clam Tridacna derasa. PMID- 15116230 TI - Fabrication of carbon microelectrodes with a micromolding technique and their use in microchip-based flow analyses. AB - In this paper, we report a new technique to pattern carbon microelectrodes for use in microfluidics. This technique, termed micromolding of carbon inks, uses poly(dimethylsiloxane)(PDMS) microchannels to define the size of the microelectrode. First, PDMS microchannels of the approximate dimensions desired for the microelectrode are made by soft lithography. The PDMS is then reversibly sealed to a substrate and the microchannels are filled with carbon ink. After a heating step the PDMS mold is removed, leaving a carbon microelectrode with a size slightly smaller than the original PDMS microchannel. The resulting microelectrode (27 microm wide and 6 microm in height) can be reversibly sealed to a PDMS-based flow channel. Fluorescence microscopy showed that no leakage occurred around the chip/electrode seal, even up to flow rates of 10 microL min( 1). The electrode was characterized by microchip-based flow injection analysis. Injections of catechol in Hank's Balanced Salt Solution (pH 7.4), showed a linear response from 2 mM to 10 microM (r(2)= 0.995), with a sensitivity of 56.5 pA microM(-1) and an estimated limit of detection of 2 microM (0.27 picomole, S/N=3). Reproducibility of the electrode response was shown by repeated injections (n= 10) of a 500 microM catechol solution, resulting in a RSD of 4.6%. Finally, selectivity was demonstrated by coating the microelectrode with Nafion, a perfluoronated cation exchange polymer. Dopamine exhibited a response at the modified microelectrode while ascorbic acid was rejected by the Nafion-coating. These electrodes provide inexpensive detectors for microfluidic applications while also being viable alternatives to use of other carbon microelectrode materials, such as carbon fibers. Furthermore, the manner in which the microelectrodes are produced will be of interest to researchers who do not have access to state of the art microfabrication facilities. PMID- 15116231 TI - Sub-micrometre imaging of metal surface corrosion by scanning Kelvin nanoprobe. AB - The detection of the initial changes in the surface microstructure and local chemical properties connected to the corrosion process is possible using a high resolution scanning Kelvin nanoprobe. This technique provides the simultaneous imaging of the topographical features and potential distribution across a surface at the sub-micrometer level. Nanoprobe measurements performed on samples of Al, Cu, Fe, Ni, Ag and Pt before and after exposure to different corroding solutions reveal the significant changes that take place during the first stages of the corrosion process. The similarities of the images obtained subsequent to corrosion are reflective of surface fractal behavior. PMID- 15116232 TI - Quaternary ammonium substituted agarose as surface coating for capillary electrophoresis. AB - A novel positively charged polymer of quaternary ammonium substituted agarose (Q agarose) has been synthesized and explored for use as a coating in capillary electrophoresis. The fast and simple coating procedure is based on a multi-site electrostatic interaction between the polycationic agarose polymer and the negatively charged fused-silica surface. By simply flushing fused-silica capillaries with hot polymer solution a positively charged, hydrophilic deactivation layer is achieved. The polymer surface provides an intermediate electroosmotic flow of reversed direction, over a range of pH 2-11, compared to unmodified fused-silica. The coating procedure was highly reproducible with an RSD of 4%, evaluated as the electroosmotic flow mobility for 30 capillaries prepared at 10 different occasions. The application of Q-agarose coated capillaries in separation science was investigated using a set of basic drugs and model proteins and peptides. Due to the intermediate electroosmotic flow generated, the resolution of basic drugs could be increased, compared to using bare fused-silica capillaries. Moreover, the coating enabled separation of proteins and peptides with efficiencies up to 300.000 plates m(-1). PMID- 15116233 TI - Analysis of size-segregated aerosol-bound polycyclic aromatic hydrocarbons with high time resolution using two-step laser mass spectrometry. AB - Polycyclic aromatic hydrocarbons are known for their mutagenic and carcinogenic properties. They are mainly emitted into the atmosphere by anthropogenic, incomplete combustion sources. Their trends over the course of a day are of interest in air quality management. A new combination of methods has been developed for the qualitative monitoring of polycyclic aromatic hydrocarbons in ambient aerosols with high time and size resolution. This has been accomplished by combining sampling with a rotating drum impactor and the analysis two-step laser mass spectrometry (L2MS). A validation for this method was carried out. Essential features of these combined techniques are (i) continuous, automatic sampling and (ii) selective as well as sensitive analysis due to the low detection limits which can be achieved with the L2MS analysis instrument. Analysis of a field sample taken in downtown Zurich, Switzerland, underlines the usefulness of this combined method, for example for following diurnal cycles of polycyclic aromatic hydrocarbons. PMID- 15116234 TI - Capturing of acidic macromolecules from biological samples using a temperature responsive polymer modified with poly-l-lysine. AB - We synthesized a temperature-responsive polymer, N-(isopropylacrylamide) methacrylic acid copolymer, to which poly-l-lysine was introduced. The synthesized polymer as well as the parent polymer showed reversible soluble insoluble changes in response to temperature changes across the lower critical solution temperature at 32 degree C in an aqueous solution. We found that the polymer efficiently captured acidic bio-macromolecules such as RNA, glycosaminoglycans and mucin-type glycoproteins in biological samples, and the captured molecules were recovered using aqueous NaCl solutions at high concentration. The target acidic molecules thus obtained will be employed for further studies such as structural analysis after brief desalting procedure. The proposed method does not require any chromatographic separations, but only needs a small volume of an aqueous salt solution for releasing captured molecules. Overall procedures are quite easy and simple, and are completed at least within 1 h. We show a few examples for capturing RNA and glycosaminoglycans from cultured cells using the polymer. PMID- 15116235 TI - Amperometric detection of glucose using self-catalytic carbon paste electrodes. AB - The analytical detection of d-glucose by means of self-catalytic carbon paste electrodes is described. In the construction of these electrodes, carbon powder has been modified with the redox liquid n-butylferrocene, which simultaneously serves both to help bind the paste together whilst also acting as a mediator in the enzymatic oxidation of d-glucose by glucose oxidase. The sensor then functions by monitoring the electrochemical oxidation of the constituent n butylferrocene itself. Through testing in model glucose solutions, the electrodes were found to yield a linear response over a d-glucose concentration range of 2 20 mM. They were also successfully employed in the determination of d-glucose levels in a spiked blood sample, giving a detection limit of 0.8 mM (based on the 3sigma criterion). PMID- 15116236 TI - QCM array for on-line-monitoring of composting procedures. AB - Six QCM resonators forming a sensor array were coated with different molecularly imprinted polymers for the on-line monitoring of composting procedures. Four key analytes are traced, namely water, 1-propanol, ethyl acetate and limonene. Trendlines obtained on-line by the sensor during measurements in a commercial composter give a distinct pattern: the signal for the alcohols first decreases according to an increase in ethyl acetate concentration, and increases again, when obviously no more acetic acid is formed. Limonene is detected in later stages of composting. Similar trends could also be observed by GC-MS. Additionally, chromatographic and sensor data for limonene could be correlated with each other. PMID- 15116237 TI - Matrix effects in immunobiosensor determination of clenbuterol in urine and serum. AB - The construction of immunochemical inhibition assays for beta-agonist and hormone residues have previously been described. In the present work the beta-agonist assay was further optimised for application to biological samples, using urine as the main model matrix. Matrix interferences with the antigen-antibody interaction and non-specific binding (NSB) of matrix components to the sensor surface were systematically studied. A full factorial design experiment was employed for evaluating the effects of assay buffer composition. In addition, the influence of antibody concentration and sample dilution on the matrix background was investigated. NSB from urine was highly affected by buffer pH and salt concentration, while buffer composition had little effect on matrix interferences with the antigen-antibody interaction. Ultra-filtration efficiently prevented NSB from urine and serum samples. Increased antibody dilution reduced the matrix background while sample dilution had an opposite effect. PMID- 15116238 TI - Scanning electrochemical microscopy imaging of rhodochrosite dissolution using gold amalgam microelectrodes. AB - Gold/mercury amalgam (Au/Hg) microelectrodes with a diameter of 25 microm were developed for the detection of environmentally relevant analytes such as manganese and iron by scanning electrochemical microscopy (SECM), and applied to investigate the controlled dissolution of manganese carbonate (MnCO(3); rhodochrosite) in acidic conditions. Characterization of the amalgam electrode geometry via approach curves recorded during SECM experiments revealed Au/Hg microelectrodes with sphere cap geometry. Quantitative determination of Mn(2+) has been achieved by calibration of the Au/Hg microelectrode in bulk solution experiments. Subsequent SECM imaging experiments confirm the applicability of amalgam microelectrodes for imaging of Mn(2+) production during dissolution of MnCO(3) at pH 3.9. This study confirms feasibility and provides the fundamental basis of SECM imaging with amalgam microelectrodes to address biogeochemically relevant questions. PMID- 15116239 TI - Reference materials for analytical toxicology including doping control: freeze dried urine samples. AB - The feasibility of freeze-dried urine samples containing doping agents to be used in intercomparison exercises and/or as reference materials has been evaluated. Freeze-dried urine samples containing caffeine, ephedrine derivatives (ephedrine, methylephedrine, norephedrine, pseudoephedrine and norpseudoephedrine), amphetamine derivatives (amphetamine, metamphetamine, 3,4 methylenedioxyamphetamine and 3,4-methylenedioxymethamphetamine) and testosterone and epitestosterone glucuronides have been evaluated. For preparation of the samples, blank urines previously subjected to filtration for clarification were fortified with standard solutions of the corresponding compounds and filtered under sterile conditions. Some aliquots of the sterile liquid samples were used for homogeneity testing, others were stored at -80 degree C for reference purposes, and the rest were subjected to lyophilisation. Freeze-dried urine samples were stored at 4-8 degree C and their stability was assessed for a period up to 18 months. Results obtained showed minimal differences (lower than 5%) between lyophilised and non-lyophilised aliquots (stored at -80 degree C) at all time periods except for amphetamine (up to 18%) and norpseudoephedrine (up to 10%). Nevertheless, such differences remained constant over the entire period of study, indicating that the loss of analytes was due to the initial lyophilisation process. The loss of analytes in freeze-dried samples was due to their volatility. Furthermore, an increase in pH by 1 unit was observed following reconstitution of samples prepared from drug-free urine of commercial origin. PMID- 15116240 TI - The roles of template complexation and ligand binding conditions on recognition in bupivacaine molecularly imprinted polymers. AB - A model for the molecular basis for ligand recognition in bupivacaine imprinted methacrylic acid-ethylene glycol dimethacrylate co-polymers has been developed based upon a series of (1)H-NMR studies in conjunction with HPLC and radioligand binding analyses. (1)H-NMR studies indicated that functional monomer-template complexes survive the polymerisation process, at least up until the stage of gelation. Polymers were synthesised and characterised by surface area analysis (BET), FT-IR and SEM. A combination of zonal and frontal chromatographic studies in aqueous and non-polar media indicate that selectivity arises from a combination of hydrophobic and electrostatic interactions. However, in the concentration regime employed for LC-based studies, ligand recognition in aqueous media was shown to be predominantly non-specific and hydrophobic in character. Radioligand binding studies, in lower ligand binding concentration regimes, permitted closer examination of the higher affinity binding sites. It was shown that the presence of a polar modifier in a non-polar solvent, or an organic modifier in water, produced enhanced selectivity. Variable temperature studies showed that the temperature of binding influences selectivity as well as the apparent number of sites available and that this effect is different in organic and aqueous environments. This indicates that the system studied is more complex in character than is generally appreciated. A comparison of the techniques employed here indicates that although chromatographic studies provide a valuable first-round screen for polymer-ligand selectivities, the level of detail obtainable using radioligand binding studies (lower concentrations and true equilibrium binding) makes them superior for detailed evaluations of molecularly imprinted polymers. PMID- 15116241 TI - Monitoring of a heterogeneous reaction by acoustic emission. AB - The feasibility of monitoring the reaction of itaconic acid and 1-butanol by non invasive acoustic emission measurements has been assessed. A piezoelectric transducer with a resonant mode at 90 kHz was attached to the external wall of a 1 L jacketed glass reactor. Acoustic emission from the oil jacket, stirrer and toluene was insignificant in comparison to that produced by the itaconic acid particles, which was transmitted through the glass walls and heating oil to the transducer. The transducer responded to acoustic emission from itaconic acid up to approximately 300 kHz, with the region around 90 kHz having the highest sensitivity. The effect of particle concentration and size on the acoustic emission generated has also been investigated, with higher concentrations and larger particles giving the greater signals. The detection limit for itaconic acid particles was 14 g dm(-3) of toluene. The effect of 1-butanol concentration and temperature on the progression of reactions was monitored using acoustic emission. It was possible to detect differences in the rate and extent of the reaction under different conditions, and also to identify when a combination of the concentration and/or size of itaconic acid particles had reached a steady state. However, it was not possible to differentiate between changes in particle size and concentration using the resonant transducer. PMID- 15116242 TI - Flow-injection determination of amine contaminants in cyclamate samples based on temperature for controlling selectivity. AB - This paper describes a flow-injection (FI) method for the simultaneous determination of aniline and cyclohexylamine impurities in cyclamate products. The method consists of the derivatization of amines with 1,2-naphthoquinone-4 sulfonate under selective and non-selective conditions. Here, the selectivity is achieved by working at 20 degree C, at which only aniline reacts, whilst higher temperatures (80 degree C) lead to a non-selective reaction of the two analytes. The FI manifold is composed of two flow cells for the spectrophotometric detection of derivatives at 480 nm. Experimental conditions have been optimized by factorial design and multicriteria making approach. Quantification is accomplished by differential analysis of the analyte contributions in the double peaks generated when the sample reaches cell 1 and cell 2. Results obtained with the proposed method are in satisfactory agreement with those provided by the standard method for the analysis of cyclamate samples. PMID- 15116243 TI - Thrombosis and nitric oxide donor drugs. PMID- 15116244 TI - A review of the therapeutic uses of thrombin. AB - Thrombin is the product of the hemostatic response essential to the conversion of fibrinogen to fibrin. In addition, it is also responsible for the aggregation of blood platelets in the formation of the "platelet plug" as well as the activation of factor VIII, factor V, factor XI, factor XIII and protein C. The action of thrombin is not confined to the hemostatic response as it also has a critical function in the wound healing process by stimulating 'mitogenic' events through interaction with cell surface receptors. In this review, we consider the various biological activities of thrombin as they relate to current therapeutic use. While there has been considerable interest in the development of fibrin sealant products, there has been considerably less interest in documenting the continuing use of thrombin as a therapeutic. The use of thrombin for topical hemostasis and the treatment of pseudoaneurysms will be discussed in detail. It is concluded that the use of thrombin as a drug will not only continue but also will significantly increase. However, the availability of a safe human thrombin preparation will be critical for the continued use of thrombin as a therapeutic. PMID- 15116245 TI - Plasminogen activator inhibitor-type 1: its plasma determinants and relation with cardiovascular risk. AB - The habitual level of PAI-1 is influenced by many factors, of which obesity and insulin resistance are the most important. It is possible to reduce plasma PAI-1 by changes in life style, e.g. weight reduction and physical activity. Data on potential interactions between environmental and metabolic variables on one hand, and the 4G/5G-polymorphism on the other hand, are still scarce. It becomes more and more clear that PAI-1 may possibly not be a major (causal) factor in cardiovascular disease, but its role in inflammation deserves further attention. In the presence of the 4G-allele not only the PAI-1 response was more pronounced, but also the response of other acute-phase reactants, which implies that the increases of these reactants are secondary to the increase in PAI-1. A myocardial infarction also provokes an acute phase response. It can thus be hypothesized that the 4G-allele might exacerbate tissue injury during the acute phase after a myocardial infarction, and thereby negatively affect the prognosis. PMID- 15116246 TI - Injection of recombinant activated factor VII can induce transient increase in circulating procoagulant microparticles. AB - Recombinant activated factor VII (rFVIIa) is an effective haemostatic treatment in haemophiliacs with inhibitors. In vitro, FVIIa concentrations corresponding to those obtained with therapeutic doses of rFVIIa have been shown to induce normal thrombin generation and platelet activation in the absence of factors VIII or IX. To further study the in vivo haemostatic changes induced by rFVIIa, circulating procoagulant microparticles (MP) were measured in patients treated with discontinuous injections of Novoseven. In 6 out of 15 patients, a transient peak of procoagulant MP was observed after injection, occurring 15 min to 2 h after infusion. It was composed primarily of platelet-derived MP and was of very short duration. This peak was not observed in haemophiliacs without inhibitor, who were treated with conventional replacement therapies. Our results provide further in vivo evidence that rFVIIa specifically activates platelets, either directly or as a consequence of a burst of thrombin generation that could account for its haemostatic efficacy. PMID- 15116247 TI - AGEs, macrophage colony stimulating factor and vascular adhesion molecule blood levels are increased in patients with diabetic microangiopathy. AB - In vitro experiments and animal models indicate that advanced glycation end products (AGEs) may play a crucial role in the vascular dysfunctions observed in patients with diabetes mellitus. These results prompted us to study subrogate markers of inflammation or vascular dysfunction in type II diabetic patients. Monocyte count and activation are dependent upon macrophage colony stimulating factors (M-CSF). Soluble vascular cell adhesion molecule (sVCAM-1) blood levels have been proposed as a marker for endothelium activation. To explore a possible relationship between these factors in diabetic patients, we measured a chemically defined AGE, N(carboxymethyl)lysine-protein (CML-protein) in a group of normal subjects (n = 55) and of diabetic patients (n = 40) using ELISA. Simultaneously, we determined M-CSF and sVCAM-1 blood levels. We found that CML-protein blood levels were significantly higher in patients with diabetes compared to non diabetic subjects (40.2 +/- 4.7 and 7.9 +/- 0.7 pmol/mg protein respectively, p < 0.0001). M-CSF was increased while sVCAM-1 blood levels were normal in the group of diabetics. M-CSF blood level was correlated to CML-protein blood level (p < 0.05). In addition CML-protein, M-CSF and sVCAM-1 were increased in patients with micro-angiopathy. These results suggest that AGE may contribute to vascular dysfunction including microangiopathy. PMID- 15116248 TI - Penthalaris, a novel recombinant five-Kunitz tissue factor pathway inhibitor (TFPI) from the salivary gland of the tick vector of Lyme disease, Ixodes scapularis. AB - Tick saliva is a rich source of molecules with antiinflammatory, antihemostatic and immunosupressive properties. In this paper, a novel tick salivary gland cDNA with sequence homology to tissue factor pathway inhibitor (TFPI) and coding for a protein called Penthalaris has been characterized from the Lyme disease vector, Ixodes scapularis. Penthalaris is structurally unique and distinct from TFPI or TFPI-like molecules described so far, including Ixolaris, NAPc2, TFPI-1 and TFPI 2. Penthalaris is a 308-amino-acid protein (35 kDa, pI 8.58) with 12 cysteine bridges and 5 tandem Kunitz domains. Recombinant Penthalaris was expressed in insect cells and shown to inhibit factor VIIa (FVIIa)/tissue factor(TF)-induced factor X (FX) activation with an IC50 of approximately 100 pM. Penthalaris tightly binds both zymogen FX and enzyme FXa (exosite), but not FVIIa, as demonstrated by column gel-filtration chromatography. At high concentrations, Penthalaris attenuates FVIIa/TF-induced chromogenic substrate (S2288) hydrolysis and FIX activation. In the presence of DEGR-FX or DEGR-FXa, but not des-Gla-DEGR FXa as scaf-folds, tight and stoichiometric inhibition of FVIIa/TF was achieved. In addition, Penthalaris blocks cell surface-mediated FXa generation by monomer (de-encrypted), but not dimer (encrypted) TF in HL-60 cells. Penthalaris may act in concert with Ixolaris and other salivary anti-hemostatics in order to help ticks to successfully feed on blood. Penthalaris is a novel anticoagulant and a tool to study FVIIa/TF-initiated biologic processes. PMID- 15116249 TI - Association after linkage analysis indicates that homozygosity for the 46C-->T polymorphism in the F12 gene is a genetic risk factor for venous thrombosis. AB - In a family-based study called GAIT (Genetic Analysis of Idiopathic Thrombophilia) that included a genome-wide scan we demonstrated that a polymorphism (46C-->T) in the F12 locus jointly influences variability of plasma (Factor XII) FXII levels and susceptibility to thrombotic disease. It then became germane to determine the prevalence of the 46C-->T polymorphism and its relative risk of thrombotic disease. We followed up evidence for genetic linkage with a case-control study, including 250 unrelated consecutive Spanish patients suffering from venous thrombotic disease and 250 Spanish subjects matched for sex and age as a controls. We measured FXII levels and genotyped the 46C-->T polymorphism, as well as a number of classical risk factors for thrombotic disease. We confirmed that individuals with different genotypes for this polymorphism showed significant differences in their FXII levels. Most importantly, the mutated T allele in the homozygous state (genotype T/T) was associated with an increased risk of thrombosis (adjusted OR of 4.82; 95% CI 1.5 15.6), suggesting that the polymorphism itself is an independent risk factor for venous thromboembolism. This study confirms that the 46C-->T polymorphism is a genetic risk factor for venous thrombosis in the Spanish population. In addition, our results confirm that a genome-wide scan coupled with a classical case-control association study is an extremely valuable approach to identify DNA variants that affect complex diseases. PMID- 15116250 TI - Contribution of polymorphisms in the endothelial protein C receptor gene to soluble endothelial protein C receptor and circulating activated protein C levels, and thrombotic risk. AB - Endothelial cell protein C receptor (EPCR) enhances the generation of activated protein C (APC) by the thrombin-thrombomodulin complex. A soluble form of EPCR (sEPCR), which is generated by metalloprotease activity, is present in plasma. The distribution of sEPCR levels in healthy populations is bimodal. Previously, we described two polymorphisms in exon 4 of the EPCR gene, 4600A/G that encodes the substitution of Ser219 by Gly in the transmembrane region of EPCR and 4678G/C in the 3'-UT region. The aim of this study was to investigate the relationship between these two polymorphisms and plasma sEPCR and APC levels and risk of venous thrombosis. We genotyped 401 healthy controls from the Spanish population and measured their plasma sEPCR and APC levels. Carriers of the 4600AG genotype had significantly higher sEPCR levels than those with the AA genotype, while the 4678CC genotype was associated, to a lesser extent, with elevated APC levels. To assess the effect of these polymorphisms on the risk of thrombosis, we genotyped 405 patients with venous thromboembolism. The frequency of the 4600AG genotype was very similar in patients and controls (p=0.975), whereas the 4678CC genotype was significantly more frequent in controls than in patients (p=0.008). In multivariate analysis, carriers of the 4678CC genotype had a decreased risk of thrombosis (OR=0.61, p=0.009). These data indicate that individuals carrying the 4600AG genotype have high sEPCR levels but do not have an increased risk of thrombosis, whereas individuals carrying the 4678CC genotype have higher APC levels and lower risk of venous thromboembolism. PMID- 15116251 TI - The anticoagulant action of recombinant human activated protein C (rhAPC, Drotrecogin alpha activated): comparison between cord and adult plasma. AB - The present study was performed to compare the anti-coagulant efficiency of recombinant human activated protein C (rhAPC) in cord with that in adult plasma. RhAPC is a promising candidate to improve the outcome of severe sepsis. However, different anticoagulant efficiency of rhAPC in cord compared with adult plasma has to be expected due to physiological low plasma levels of tissue factor pathway inhibitor (TFPI) and antithrombin (AT) present in neonates, two inhibitors known to markedly influence the anticoagulant action of APC. Clot formation was induced in our experiments by addition of high (30 micro M) or low (20 pM) amounts of lipidated tissue factor (TF). High amounts of TF are conventionally applied in standard clotting assays, whereas plasma activation with low amounts of TF probably better matches the conditions in vivo. We demonstrate that under low coagulant challenge increasing amounts of rhAPC (0.1 0.5 micro g/ml final plasma concentration) dose-dependently prolonged clotting time and suppressed thrombin potential and prothrombin fragment 1+2 generation in both cord and adult plasma. The same was true for experiments performed under high coagulant challenge when 4-16 micro g/ml of rhAPC were added. Whereby, cord plasma was significantly more susceptible to addition of rhAPC in the presence of high amounts of TF and adult plasma was significantly more susceptible to addition of rhAPC in the presence of low amounts of TF. We demonstrate that increased anticoagulant efficiency of rhAPC in adult plasma under low coagulant challenge is attributable to the physiological high levels of TFPI and AT present in adults. PMID- 15116252 TI - Orally administered heparins prevent arterial thrombosis in a rat model. AB - Our previous studies demonstrated that orally administered heparins prevent thrombosis in a rat jugular vein thrombosis model, where bovine unfractionated heparin (UFH) and the low molecular weight heparin tinzaparin reduced thrombotic incidence by 50% at 7.5 and 0.1 mg/kg, respectively. Our objectives were to determine if similar antithrombotic effects of oral heparin could be observed in an arterial thrombosis model. In this model, filter paper soaked in 30% ferric chloride was applied to the exposed rat carotid artery. A flowmeter recorded blood flow over a 60 min period determining time when the thrombus began forming (TTB) and time till occlusion (TTO). Immediately following, the thrombus was removed, dried and weighed 24 h later. Bovine UFH (7.5 mg/kg), tinzaparin (0.1 mg/kg) or saline was administered by stomach tube at 2, 5 and 25 h prior to thrombus initiation. TTB was significantly increased when UFH was given at 5 and 25 h but not 2 h prior, and when tinzaparin was given at 5 but not 2 or 25 h prior compared to rats given oral saline. TTO was significantly increased for both UFH and tinzaparin when given 5 and 25 h but not 2 h prior (one-way ANOVA). There was no difference in TTO and TTB between UFH and tinzaparin treated groups. A trend in reduction in thrombus weight was observed for UFH at 5 and 25 h prior and tinzaparin at 5 h prior to thrombus initiation (one-way ANOVA). Although no significant changes were observed in activated partial thromboplastin times, Heptest or anti-Xa activity from plasma of heparin treated rats, endothelial heparin concentrations were significantly greater than controls for UFH at 5 h and for tinzaparin at 2, 5, and 24 h. Thus, heparins administered by the oral route are effective antithrombotic agents in arterial as well as venous models. PMID- 15116253 TI - Localization of heparin and low-molecular-weight heparin in the rat kidney. AB - Unfractionated heparin (UFH) and low-molecular-weight heparin (LMWH) are cleared, at least in part, by the kidneys through a poorly understood process. This study was undertaken to explore the mechanism of renal clearance of these drugs. Rats were given fluorescein-5-isothiocyanate (FITC)-labeled UFH or LMWH intravenously. At intervals after injection, rats were euthanized and the kidneys were harvested and subjected to immunohistochemical analysis and fluorescence microscopy. Both UFH and LMWH were localized to renal tubular cells and no immunoperoxidase staining or fluorescence was detected in glomeruli. Autoradiography demonstrated similar intracellular distribution of radio-labeled UFH suggesting that this phenomenon is independent of the method used to label heparin. Fluorescence in the tubules increased as a function of time after UFH injection, but reached a plateau after LMWH injection suggesting that the rate of renal tubular uptake depends on the molecular size of the heparin. When administered prior to FITC labeled UFH or LMWH, probenecid, a renal organic anion inhibitor, decreased the renal tubular uptake of the heparins, whereas cimetidine, a renal organic cation inhibitor, had no effect. These findings suggest that renal excretion of UFH and LMWH primarily reflects tubular uptake via an organic anion transport mechanism. PMID- 15116254 TI - Prospective evaluation of coagulation activation in pregnant women receiving low molecular weight heparin. AB - Pregnancy is a major risk factor for venous thromboembolism (VTE), and low molecular weight heparin (LMWH) seems to be safe and effective in pregnant women. Normal pregnancy is accompanied by a state of hypercoagulability, indicated by an increase in markers of coagulation activation. In a prospective cohort study, we followed 61 women who received LMWH thromboprophylaxis throughout pregnancy because of a history of VTE, hereditary thrombophilia and/or previous pregnancy related complications. The control group consisted of 113 healthy pregnant women without antithrombotics. D-Dimer, prothrombin fragment F1+2 (F1+2) and the resistance to activated protein C (APC-ratio) were measured in all women during the first, second and third trimester. Patients and controls did not significantly differ with regard to baseline characteristics and pregnancy outcome. A (recurrent) VTE was seen in one patient despite LMWH. D-Dimer levels significantly increased among patients and controls during pregnancy (p < 0.0001), and were significantly higher among patients compared with the controls (p <0.0001) [395 ng/ml (95% CI 340-458) and 249 ng/ml (95%CI 234-266); 710 ng/ml (95% CI 602-838) and 475 ng/ml (95% CI 431-523); 1089 ng/ml (95% CI 931-1273) and 822 ng/ml (95% CI 741-911); respectively]. Levels of F1+2 significantly increased while the APC-ratio significantly decreased during pregnancy among patients and controls. Despite LMWH, pregnancy is accompanied by a substantial activation of the coagulation system. PMID- 15116255 TI - Lower limb venous haemodynamic impairment on dependency: quantification and implications for the "economy class" position. AB - The role of stasis in venous disease is undisputed, yet surprisingly, its haemodynamic quantitation remains largely undefined. We investigated the phenomenon of venous stasis in the lower limb upon sitting and standing and project its implications to economy class aircraft passengers. 26 normal limbs, 13 subjects, age 29-54, selected after duplex, plethysmography and ABPI, had peak[V(peak)], mean[V(mean)] and minimum[V(min)] velocities, volume flow[Q(venous)], pulsatility index [PI(venous)] and diameter obtained on horizontal, sitting (as in economy aircraft seats) and standing with duplex, at popliteal, femoral[FV] and common femoral[CFV]veins [differences in median %]. V(peak), V(mean) and Q(venous) decreased from horizontal to sitting in the CFV [57%, 71%, 31%, respectively], FV [51%, 70%, 34%] and popliteal [31%, 58%, 42%] (all, p<.001). V(peak),V(mean) and Q(venous) decreased further from sitting to standing in the CFV [26%, 44%, 25%, respectively], FV [21%, 42%, 27%] and popliteal [14%, 42%, 20%] (all, p <.001). Diameter, V(min) and PI(venous) increased from horizontal to sitting in the CFV [50%, 63%, 38%, respectively], FV [39%, 23%, 66%] and popliteal [21%, 14%, 84%] (all, p <.001)]. Diameter, V(min) and PI(venous) increased further from sitting to standing in CFV [10%, 22%, 19%, respectively; p =.004], FV [12%, 68%, 2%[ns]; p <.001)] and popliteal [14%, 50%, 24%; p =.017]. In all postures: V(peak), V(mean), Q(venous) and diameter at CFV exceeded FV (p <.025) and popliteal (p <.001) ones; also those at FV exceeded the popli-teal ones (p =.003), except for the diameter on horizontal. V(min) in popliteal was higher than in CFV (p =.003) or FV (p <.025), on horizontal and standing. PI(venous) in CFV was lower than in FV or popliteal (p <.025) on sitting. Right to left differences non-significant. [Wilcoxon(+Bonferroni) test: significance at p <.025] A shift from horizontal to sitting generates a most significant attenuation in Q(venous),V(peak) and V(mean) linked to a reciprocal increase in V(min), PI(venous) and vein diameter, with further exacerbation on standing. V(peak),V(mean) and Q(venous) decline with distance from groin enhancing venous stasis in the periphery. By restricting activation of the natural venous pumps, sitting cramped during long flights may protract the status of haemodynamic stagnation sustained on dependency which paired with marked venous dilatation generates a milieu that may promote thrombogenesis. PMID- 15116256 TI - GPVI down-regulation in murine platelets through metalloproteinase-dependent shedding. AB - Platelet interaction with subendothelial collagen is crucial for hemostasis and thrombosis. Under conditions of elevated shear, platelet adhesion and activation on collagen requires the coordinated action of glycoprotein (GP) Ib and GPVI, which may be physically and functionally linked in the platelet membrane. While the surface expression of GPIb can be down-regulated by internalization and/or proteolytic cleavage of a 130 kDa fragment of GPIb (glycocalicin, GC), very little is known about the cellular regulation of GPVI. We have recently shown that GPIb on platelets is cleaved by metalloproteinase-dependent mechanisms in response to mitochondrial injury. In the current study, we examined a possible role of platelet metalloproteinases in the regulation of GPVI. Mitochondrial injury induced by incubation of mouse platelets with carbonyl cyanide m chlorophenyl-hydrazone (CCCP) severely affected the cells' responses to collagen or the GPVI-specific agonist, collagen related peptide (CRP), but not to thrombin or the stable thromboxane A(2) analog U46619. This defect was due to a rapid proteolytic cleavage of GPVI, as shown by the release of the 55 kDa extracellular domain into the supernatant. Both the proteolysis of GPVI and the loss of its activity were inhibited in the presence of the broad range metalloproteinase inhibitor, GM6001. Platelet stimulation with thrombin or CRP, however, resulted in marked metalloproteinase-dependent shedding of GPIbalpha, but not GPVI suggesting that different metalloproteinases are involved in the regulation of the two receptors or, alternatively, an additional signal is required to render GPVI susceptible to cleavage. PMID- 15116257 TI - The novel human platelet septin SEPT8 is an interaction partner of SEPT4. AB - Septins are a family of GTP-binding proteins, which are essential for active membrane movement such as cytokinesis and vesicle trafficking. In non-dividing cells (such as platelets and neurons) septins are implicated in exocytosis. Platelets from a SEPT5 knockout mouse showed an altered serotonin secretion and platelet aggregation, suggesting that SEPT5 is involved in secretion in platelets. Septins form complexes consisting of multiple septin polypeptides. Using the yeast two-hybrid system we had demonstrated that SEPT5 partners with SEPT8. The aim of this study was to identify other interaction partners of the human platelet septin SEPT8. Using the yeast two-hybrid system with SEPT8 as bait protein we identified the human septin SEPT4 as an interaction partner of SEPT8. The interaction between SEPT4 and SEPT8 was confirmed by immunoprecipitation. Expression analysis revealed that SEPT4 is also expressed in human platelets. Thus, SEPT4 is the third described platelet septin besides SEPT5 and SEPT8. Transmission electron microscopy of platelets revealed that SEPT8 and SEPT4 are localized surrounding alpha-granules (as it had been shown for the septin SEPT5) suggesting that the three septins may be components of the septin complex in platelets and contribute in such a way to platelet biology. Activation of platelets by agonists resulted in the translocation of SEPT4 and SEPT8 to the platelet surface indicating a possible functional role of these proteins in platelet granular secretion. PMID- 15116259 TI - The role of platelets and recombinant factor VIIa on thrombin generation, platelet activation and clot formation. AB - In the present study we assessed the effect of platelet counts and rFVIIa on thrombin generation, platelet activation and clot formation after tissue factor pathway activation in human plasma aiming to investigate the mechanism by which rFVIIa induces haemostasis in patients with severe thrombocytopenia. Plasma samples with platelet counts from 5 x 10(9)/l to 150 x 10(9)/l were spiked with rFVIIa (1 micro g/ml) or buffer. Clotting was initiated in the presence of diluted thromboplastin. Thrombin generation was assessed using the Thrombogram Thrombinoscope trade mark assay. The kinetics of platelet activation was assessed using flow cytometry to measure the expression the P-selectin on platelet membrane of washed platelets suspended in defibrinated homologous PPP. Thromboelastography was used to evaluate the effect of platelets and rFVIIa on the kinetics of clot formation and clot's firmness. In the presence of low platelet counts the endogenous thrombin potential (ETP) and the maximum concentration of generated thrombin (Cmax) were reduced by 60%-70%. The lag-time of thrombin generation and the time required to reach the Cmax (Tmax) were prolonged, the velocity of platelet activation was decreased and thrombus formation was delayed. Recombinant FVIIa accelerated thrombin generation and platelet activation but it did not significantly modify ETP or Cmax. Recombinant FVIIa enhanced platelet activation in a TF and thrombin dependent manner since its effect on the studied parameters was abolished when TF was omitted or when hirudin was added into the experimental system respectively. Recombinant FVIIa normalized the velocity of clot formation but it did not modify clot firmness, which depended mainly on platelets' count. In conclusion, in experimental conditions simulating severe thrombocytopenia rFVIIa in the presence of low amounts of TF, accelerates thrombin generation, without increasing the maximum amount of generated thrombin, thus leading in enhanced platelet activation and rapid clot formation. PMID- 15116258 TI - Strong correlation between the prevalence of cerebral infarction and the presence of anti-cardiolipin/beta2-glycoprotein I and anti-phosphatidylserine/prothrombin antibodies--Co-existence of these antibodies enhances ADP-induced platelet activation in vitro. AB - Cerebral infarction is the most common arterial thromboembolic complication in the anti-phospholipid antibodies (aPL) syndrome. In an effort to clarify the roles of aPL in the pathogenesis of cerebral infarction in patients with SLE, we examined the levels of anti-cardiolipin/2-glycoprotein I antibodies (anti CL/beta2-GPI) and anti-phosphatidylserine/prothrombin anti-bodies (anti-PS/PT) in addition to lupus anticoagulant (LA) activity in 126 patients with SLE (35 with cerebral infarction and 91 without thrombosis). Both anti-CL/beta2-GPI and anti PS/PT strongly correlated with the presence of LA activity. The prevalence of cerebral infarction was obviously higher in the patients who had both anti CL/beta2-GPI and anti-PS/PT (76.5% [26/34 cases], p<0.0001) than in the other patients having anti-CL/beta2-GPI or anti-PS/PT alone or neither of them (9.8% [9/92 cases]). Furthermore, we studied the in vitro effects of anti-CL/beta2-GPI and/or anti-PS/PT on the enhancement of platelet activation induced by stimulation with a low concentration of adenosine diphosphate (ADP). The purified IgG containing both anti-CL/beta2-GPI and anti-PS/PT caused significant enhancement of platelet activation caused by ADP. However, the purified IgG containing either anti-CL/beta2-GPI or anti-PS/PT had no enhancing effects on it. Furthermore, platelet activation was generated by the mixture of anti-CL/beta2 GPI-IgG and anti-PS/PT-IgG prepared from individual patients, but not by each fraction alone. These results indicate that anti-CL/beta2-GPI and anti-PS/PT may cooperate to promote platelet activation, which may contribute to the risk of cerebral infarction in patients with SLE. PMID- 15116260 TI - Pharmacogenetics of the CD14 endotoxin receptor polymorphism and progression of coronary atherosclerosis. AB - Atherosclerosis is at least in part an inflammatory disease. CD14 is an endotoxin receptor that after binding of lipopolysaccharides evokes endothelial activation and secretion of several cytokines. A polymorphism of CD14 has been associated with myocardial infarction. We evaluated the role of the -159 T/C polymorphism in the promoter region of the CD14 gene in relation to severity and progression of coronary atherosclerosis and response to the HMG CoA reductase inhibitor pravastatin. We recruited patients from the multi-center double-blind randomized placebo controlled REGRESS trial and genotyped the -159T/C CD14 polymorphism. DNA and angiographic follow-up were available from 759 patients with objectivated coronary artery disease. We measured changes in mean segment diameter (MSD) and minimum obstruction diameter (MOD) with quantitative coronary angiography and noted the occurrence of major adverse cardiac events. The genotype distribution was 28% TT, 49% CT, 23% CC. We did not find any association between genotype and MSD and MOD at baseline, frequency of previous myocardial infarction, changes in MSD and MOD or major clinical events. Treatment with the HMG CoA reductase inhibitor pravastatin reduced progression of coronary atherosclerosis and adverse events equally for all genotypes. We conclude, that the -159T/C polymorphism in the CD14 monocyte receptor gene was not associated with progression of coronary atherosclerosis in this population nor did it influence the efficacy of pravastatin in the treatment of atherosclerosis. PMID- 15116261 TI - Human alpha -defensins neutralize fibrinolytic activity exerted by staphylokinase. AB - Defensins, cationic peptides with bacteriolytic properties, are abundantly found at inflammation sites and in human coronary vessels. Vascular occlusive diseases, such as myocardial infarction, pulmonary embolism, and peripheral arterial occlusion are presently treated by thrombolytic intervention using staphylokinase, a plasminogen activator of bacterial origin. In this study we assessed a possible interaction between defensins and staphylokinase, both molecules being present in an acutely ill patient. Using an ELISA-based system, we found that staphylokinase and defensins displayed a strong and dose-dependent binding. In contrast, urokinase, another plasminogen activator of endogenous origin, displayed only minimal binding to defensins. Next, we proved that interaction between staphylokinase and defensins led to functional consequences resulting in a significant decrease (p<0.002) of plasminogen activation capacity upon complex formation. In contrast, urokinase retained most of its activity even in 10-fold molar excess of defensins. Finally, we found that staphylokinase triggered lysis of fibrin was efficiently inhibited in the presence of defensins. To assess structural requirements for staphylokinase/defensin interaction, six staphylokinase mutant variants were studied. Inactivation pattern of the tested staphylokinase variants suggested a direct binding of defensins to serine protease-like domain of staphylokinase. In conclusion, we show complex formation between staphylokinase and alpha-defensins resulting in a significant reduction of fibrinolytic activity. This finding may have clinical implications, since fibrinolytic effects of staphylokinase may be downregulated at the site of vascular occlusion. PMID- 15116262 TI - Influence of low frequency electric fields on anti- and pro-coagulability of the vascular endothelium: new insights into high-voltage electrical injury. AB - After high-voltage electric injury, patients often show tissue necrosis and thrombosis of blood vessels even remote from entry and exit site of electrical current. In this study, plasma levels of TAT, F(1+2), PAI-1, and t-PA were determined in vivo in three patients with high-voltage injury for 96 hours after trauma. In order to analyse a possible effect on haemostasis related to endothelial cell damage, protein S, TF, ET-1, PGI(2), NO, t-PA, and PAI-1 were determined for 72 hours in vitro in cell culture supernatant of HUVECs that had been exposed to 1, 10, 30, and 50 electric field periods of 50 Hz with field strength of 60 V/cm and duration of 20 ms. Furthermore, expression of thrombomodulin was immunohistochemically analysed. Clotting activation could be observed in our patients by increased levels of F(1+2) and TAT between 12 and 72 hours after injury, whereas fibrinolysis was disturbed due to high PAI-1. One patient presented thrombosis of vessels by day 3. In vitro, PAI-1 increased significantly (p<0.05) in medium of cells with an application of 30 and 50 periods between 2 and 48 hours. Between 4 and 72 hours, the concentration of t-PA was significantly lower (p<0.05) in the medium of HUVECs exposed to 10, 30, and 50 periods, whereas there was a significant increase (p<0.05) in the concentration of TF in the cell groups with an application of 30 and 50 periods. 24, 48, and 72 hours after injury, there was just weak or no staining for thrombomodulin in HUVECs with an application of 30 and 50 periods. The disturbed balance between clotting system and fibrinolysis seen in vitro after electric injury might explain the clinical observation of a progressive thrombosis of blood vessels after electric injury leading to tissue loss. PMID- 15116263 TI - Binding of heparin to plasma proteins and endothelial surfaces is inhibited by covalent linkage to antithrombin. AB - Unfractionated heparin (UFH) and low molecular weight heparin (LMWH) are used for prophylaxis and treatment of thrombosis. However, UFH has a short plasma half life and variable anticoagulant response in vivo due to plasma or vessel wall protein binding and LMWH has a decreased ability to inactivate thrombin, the pivotal enzyme in the coagulation cascade. Covalent linkage of antithrombin to heparin gave a complex (ATH) with superior anticoagulant activity compared to UFH and LMWH, and longer intravenous half-life compared to UFH. We found that plasma proteins bound more to UFH than ATH, and least to LMWH. Also, UFH bound significantly more to endothelial cells than ATH, with 100% of UFH and 94% of ATH binding being on the cell surface and the remainder was endocytosed. Competition studies with UFH confirmed that ATH binding was likely through its heparin moiety. These findings suggest that differences in plasma protein and endothelial cell binding may be due to available heparin chain length. Although ATH is polydisperse, the covalently-linked antithrombin may shield a portion of the heparin chain from association with plasma or endothelial cell surface proteins. This model is consistent with ATH's better bioavailability and more predictable dose response. PMID- 15116264 TI - Evidence of a net release of tissue-type plasminogen activator across the human cerebral vasculature. AB - We have earlier described models for measuring local net release rates of tissue type plasminogen activator (t-PA) in vivo across skeletal, coronary, pulmonary, and splanchnic vascular beds. Aim of the present study was to investigate whether there is a net release of t-PA across the human cerebral vascular bed and whether an acute regulated release can be induced by sympathoadrenal activation. Fourteen male subjects undergoing elective coronary artery bypass grafting were investigated prior to surgery and during sternotomy-induced sympathoadrenal activation. Blood samples were obtained simultaneously from the radial artery and the jugular bulb. Blood flow velocity in the middle cerebral artery (V(MCA)) was determined by transcranial Doppler. Cerebral net release of t-PA was calculated as the arterio-venous concentration gradient times V(MCA). Prior to surgery there was a significant cerebral net release of t-PA (131 and 42 ng/min for t-PA antigen and activity, respectively). The release was significantly induced by sternotomy (to 271 and 80 ng/min, respectively). No significant cerebral net release of plasminogen activator inhibitor type 1 (PAI-1) was detected throughout the experiment. The results show that there is a basal net release of t-PA across the human cerebral vascular bed and that sympathoadrenal activation induces a local regulated release of t-PA. PMID- 15116265 TI - Adiponectin is inversely related to plasminogen activator inhibitor type 1 in patients with stable exertional angina. AB - Adipose tissue is a secretory organ producing a variety of bioactive substances, such as adiponectin. Adiponectin has antiatherogenic properties while plasminogen activator inhibitor type 1 (PAI-1) is closely involved in the development of atherosclerosis. The relationship between adiponectin and PAI-1 in patients with coronary artery disease (CAD) has not been clarified. This study examined plasma levels of adiponectin and PAI-1 in 64 patients with stable exertional angina (SEA) and 65 patients with the chest pain syndrome (CPS). Plasma log-adiponectin levels were significantly lower in patients with SEA (0.62+/-0.08 micro g/dL) compared to those with CPS (0.86+/-0.05 micro g/dL) (p<0.0001). The plasma levels of log-PAI-1 were significantly higher in patients with SEA (1.23+/-0.18 ng/mL) compared to those with CPS (1.15+/-0.22 ng/mL) (p<0.05). Plasma log-adiponectin levels correlated negatively with diabetes mellitus (DM), body mass index (BMI), log-PAI-1 (r=-0.284, p<0.001), triglyceride (TG), and remnant-like particles cholesterol (RLP-C), and positively with high-density lipoprotein cholesterol (HDL-C) levels. Plasma levels of log-PAI-1 correlated positively with DM, BMI,TG and RLP-C levels, and negatively with HDL-C levels. Multiple logistic regression analysis identified sex, angina pectoris, and PAI-1 as independent determinants of hyperadiponectinemia (p<0.05). Adiponectin is inversely related to PAI-1. DM, BMI,TG, HDL-C, and RLP-C are common mediators between adiponectin and PAI-1, and treatment for common mediators may prevent the development of CAD by reducing PAI 1 and increasing adiponectin levels. PMID- 15116266 TI - Factor V Leiden and prothrombin G20210A mutations in young adults with cryptogenic ischemic stroke. AB - The association between factor V Leiden (FVL) and prothrombin G20210A (PT 20210) mutations and ischemic stroke remains controversial, particularly in young adults with cryptogenic stroke. Prevalence of FVL (4.1%) and PT 20210 (8.2%) mutations was assessed in 49 patients under 50 years with cryptogenic stroke and compared with controls. Odd ratio (OR) for cryptogenic stroke was 2.62 (95% CI, 0.49 13.95) for FVL and 3.75 (95% CI, 1.05-13.34) for PT 20210 and 3.28 (95% CI, 1,17 9.20) for some recognized genetic thrombophilic defect. Moreover, the OR for cryptogenic stroke in young women using oral contraceptives (OC) was 3.59 (95% CI, 1.28-10.5). When some genetic thrombophilic defect was associated with OC, the OR was much higher (OR: 14.27; 95% CI, 0.66-309.99). Our results suggest that in the Mediterranean populations the PT 20210 mutation, but not FV Leiden, is a risk factor for cryptogenic stroke in young adults. OC use is also a significant risk factor for cryptogenic stroke, which is increased in women with some genetic thrombotic risk factor. PMID- 15116267 TI - A novel anti-ischemic nitric oxide donor inhibits thrombosis without modifying haemodynamic parameters. AB - Platelets are involved in the clinical presentations of ischemic heart disease. Our objective was to study the antithrombotic effects of a new nitric oxide donor (LA419), a neutral sugar organic nitrate with a protected thiol group in its molecular structure. Animals were randomly distributed in three groups: I) oral administration of LA419 (0.9-1.8-3.6-5 mg/kg/d, 10 days); II) oral administration of standard IS-5-MN (0.9-1.8 mg/kg/d, 10 days); III) non-treated group (control). In catheterized pigs, thrombosis was studied under controlled rheological conditions by radioisotopic evaluation of deposited platelets on damaged vessel wall, placed in an extracorporeal perfusion chamber. Changes in blood pressure, heart rate, and platelet aggregation were evaluated. Results have shown that LA419 significantly decreased thrombus formation according to the degree of vascular damage, and shear rate conditions in a dose-dependent manner (p<0.005), without significant modifications on blood pressure and/or elevation of liver enzymes. In contrast, IS-5-MN only showed a significant reduction on platelet deposition at the high dose, that was associated to hypotension and elevation of liver enzymes. Therefore, we conclude that this new anti-ischemic NO-donor (NOd) LA419 that inhibits platelet function without modifying blood pressure may be a highly efficacious strategy to passivate platelet activation induced by a damaged vessel wall. PMID- 15116268 TI - Lepirudin therapy for thrombotic complications in congenital afibrinogenaemia. PMID- 15116269 TI - Intravenous Iloprost treatment for severe bone pain caused by sickle cell crisis. PMID- 15116270 TI - Mechanism of action of drotrecogin alfa activated (rhAPC). PMID- 15116271 TI - PPARalpha deficiency does not modify age dependency but prevents high fat diet increase in plasma PAI-1 as well as insulin resistance. PMID- 15116272 TI - Absence of thromboembolic complications in patients with hereditary bleeding disorders undergoing major orthopaedic surgery without antithrombotic prophylaxis. PMID- 15116273 TI - Genetic diversity between Japanese and Chinese threeline grunt (Parapristipoma trilineatum) examined by microsatellite DNA markers. AB - Threeline grunt (Parapristipoma trilineatum) distributes around the southwestern coast of Japan and the east coast of China. The Chinese P. trilineatum was imported by Japan as an aquacultural seed because of its rapid growth compared with that of the Japanese P. trilineatum. The Japanese P. trilineatum differs from the Chinese P. trilineatum in some quantitative traits, and it has been suggested that these two P. trilineatum populations are genetically different. In order to identify the population structures around Japan and China, 5 local populations of the Japanese P. trilineatum and 2 local populations of the Chinese P. trilineatum were analyzed using 4 microsatellite DNA markers. Significant differences were detected between Japanese and Chinese P. trilineatum and among samples of Chinese P. trilineatum; however, among the samples of Japanese P. trilineatum, no significant differences were detected. These results suggest that care must be taken to prevent the escape of the Chinese P. trilineatum from culture cages around the Japanese coast, in order to preserve the genetically different population structures of Japanese and Chinese P. trilineatum. PMID- 15116275 TI - A study on patients treated with polyacrylamide hydrogel injection for facial corrections. AB - Polyacrylamide hydrogel (PAAG) has been used as a tissue filler in facial corrective surgery and for breast augmentation in Kiev, Ukraine, for more than 10 years with reportedly very good results. These results, however, have not been published in peer-reviewed journals. A Danish/Swedish group of plastic surgeons with special interest in facial corrective surgery did a retrospective, systematic, pre-planned investigation of 104 patients treated at the center in Kiev. All data were entered into a pre-programmed database for data processing. The mean age of this population was 37.4 years and the mean time since the gel injection was 3.9 years. An average of 5.7 ml of PAAG was injected prior to the investigation. The gel was well tolerated and assessment of the outcome was judged to be very good by 78% and good by 22%, by both physicians and patients. It is concluded that PAAG is well tolerated and seems to be a promising product for facial corrective surgery. Currently, the product (Aquamid) is being studied in several prospective clinical trials, one of which is completed and in the process of preparation for publication. PMID- 15116276 TI - Postoperative nausea and vomiting in the plastic surgery patient. AB - Despite great advances in modern medicine, postoperative nausea and vomiting (PONV) continue to be a significant problem without a definitive cure. This article describes a prospective study conducted with 143 plastic surgery patients at a single institution between 1998 and 2000 to identify preoperative risk factors for PONV that may be specific to plastic surgery patients. In addition, a prophylactic regimen was identified that demonstrated a decrease in the incidence of PONV from the published incidence of 22% to 3%. The regimen includes the use of multiple antiemetic agents including ondansetron (Zofran) promethazine (Phenergan), and metoclopramide (Reglan), with the addition of dexamethasone (Decadron) in selected cases PMID- 15116277 TI - Bone chips and diced cartilage: an anatomically adopted graft for the nasal dorsum. AB - Since 1996, cranial bone chips or septal bone chips harvested during septal deviation surgery and small chips of ear or septal cartilage have been used in 67 patients for dorsal nasal augmentation or for smoothing dorsal nasal irregularities. In this study, 59 overresections of ostecartilaginous nose structures during previous aesthetic nose surgeries and 8 primary rhinoplasties occasioned the use of bone or cartilage grafts. For 57 patients both bone and ear cartilage grafts were used for the reconstruction. Bone grafts were used for seven cases and cartilage grafts for three cases. The results from 7 years, of experience with this method of nasal dorsum reconstruction were satisfactory and durable. The most important advantage of this method is that the bony side of the nose is reconstructed with bone and the cartilage side with cartilage. Another advantage is that the bone chips are incorporated with both nasal bones, building a strong dorsal nasal bony mono-black. This technique also is useful for augmenting mild saddle nose deformity and dorsal nasal projection deficiency on the bony part, cartilage part, or both parts. PMID- 15116278 TI - Transdermal scopolamine use for post-rhytidectomy sialocele. AB - BACKGROUND: Post-rhytidectomy sialocele is a rare complication. The usual treatment uses prolonged drainage. The use of antisialogogues has been described previously, but has potential side effects. Transdermal scopolamine SCOPODERM TTS is indicated for the prevention of motion sickness and also has antisialogugue action. METHODS: An interventional case report is presented. RESULTS: The application of transdermal scopolamine SCOPODERM TTS resulted in resolution of a post-rhytidectomy sialocele within 6 days. CONCLUSION: Transdermal scopolamine delivery should be considered as an option for the treatment of post-rhytidectomy parotid fistulas. PMID- 15116279 TI - Bacterial competition in activated sludge: theoretical analysis of varying solids retention times on diversity. AB - A mechanistic model for activated sludge sewage treatment was developed to predict exploitative competition of six aerobic heterotrophic bacterial species competing for three essential resources. The central hypothesis of the model is that in a multispecies/limiting resource system the number of coexisting bacterial species, N, exceeds the number of limiting resources, K, available for them. The explanation for this is that for certain species combinations, the dynamics of the competition process generate oscillations in the abundances of species, and these oscillations allow the coexistence of greater number of species than the number of limiting resources ( N > K). This result is a direct contradiction of an existing activated sludge steady state competition theory, "the principle of competitive exclusion," which states that the competition process proceeds to equilibrium, allowing only N 5000 copies/mL, previous therapy, and very poor adherence to the medication regimen. RESULTS: Eighty-five patients were included (mean viral load, 4.4 log/mL; mean CD4, 240 cells/mL; IDUs, 78%; methadone maintenance program, 42%; AIDS, 28%). Number of previous therapies: one, 53%; two, 28%; three or more, 19%. In the intent-to-treat analysis at 1 year, 38 patients (44.7%) achieved viral load below 500 copies/mL. Adherence greater than 90% of prescribed drugs was reported in 49% of patients, adverse events were reported in 17.6%, mortality in 6%, and lost to follow-up in 26%. The factors associated with virologic failure were nonadherence (odds ratio [OR], 4.4; 95% CI 1.5-12.3), baseline CD4 cell count <200 cells/mL (OR, 3.4; 95% CI 1.3 8.9; p =.01), and more than one previous treatment (OR, 2.7; 95% CI 1.1-6.9). CONCLUSION: Regarding previously very nonadherent patients, this simplified combination therapy containing three NRTIs obtained satisfactory results in ART experienced patients. However, more aggressive interventions to enhance adherence are needed to improve results. PMID- 15116282 TI - Measuring adherence to antiretroviral therapy in a diverse population using a visual analogue scale. AB - PURPOSE: To examine the performance of an instrument to assess adherence based on a visual analogue scale, compared to an instrument based on 3-day recall, using unannounced pill counts in the place of residence as the gold standard. METHOD: We prospectively assessed adherence to antiretroviral therapy in 84 marginally housed indigent HIV-infected patients who were receiving stable antiretroviral therapy in San Francisco, California, with three adherence assessments over no more than 4 months. RESULTS: Mean adherence using the visual analogue scale, 3 day recall, and unannounced pill count methods were 82.5%, 84.2%, and 75.9%, respectively. The correlation between visual analogue scale and unannounced pill count was high (r = 0.76) and was not statistically different from that between 3 day recall and unannounced pill count (r = 0.71; p =.52). Both methods were also similarly inversely correlated with HIV viral load (r = -0.49 and -0.34, respectively; p =.22 for the difference in the correlations). The visual analogue scale correlation with unannounced pill count was stable over time and remained high in all subpopulations examined. CONCLUSION: A visual analogue scale to assess adherence was performed as well as a more complicated 3-day recall instrument in this diverse population. Given its simplicity, the visual analogue scale adherence instrument will be useful in research and may be useful in routine patient care. PMID- 15116283 TI - Interferon-gamma therapy activates human monocytes for enhanced phagocytosis of Mycobacterium avium complex in HIV-infected individuals. AB - Defective immunological function of cells of the macrophage lineage contributes to the pathogenesis of HIV-1 infection. Because monocyte/macrophage function is enhanced by cytokines such as interferon-gamma (IFN-gamma), the use of this immunomodulator is of potential clinical interest as adjunctive immunotherapy in immunosuppressed individuals. In this study, we show that adjunctive IFN-gamma treatment in an HIV-infected individual with Mycobacterium avium complex (MAC) infection increased phagocytosis of MAC by blood monocytes when compared to cells from an HIV-infected patient who was receiving standard chemotherapy alone. Enhanced phagocytic efficiency resulting from IFN-gamma therapy was associated with increased surface expression of MHC II (HLA-DR), a phagocytic receptor (CD16), and the activation marker (CD69), although the levels of activation markers were dissimilar at baseline in the two participants. These results imply that IFN-gamma may be useful in restoring antimycobacterial function in immunosuppressed patients. PMID- 15116284 TI - Benfluorex as a therapeutic option for insulin resistance in HIV lipodystrophy syndrome. AB - BACKGROUND: HIV lipodystrophy syndrome, characterized by a significant excess of visceral adiposity and a reduced subcutaneous fat mass in association with insulin resistance and dyslipidemia, still affects the majority of antiretroviral treated HIV-infected patients. The therapeutic management of this syndrome has not yet been well established. Benfluorex is known to decrease insulin resistance with no side effects on lactate levels in HIV-negative patients. METHOD: We conducted an open-label study of benfluorex (150 mg, 2-3 times a day) that was prescribed for 60 HIV-infected patients who were diagnosed with glucose metabolism abnormalities by oral glucose tolerance test (OGTT); 47 of these patients had visceral fat accumulation measured by computed tomography (VAT). Median follow-up was 12 months (interquartile range [IQR] = 6-12 months). The great majority of patients (90%) were treated with at least triple therapy (in 70% the therapy included at least one PI), with a nonsignificant change over the study period. RESULTS: Added to antiretroviral therapy, benfluorex improved OGTT in 47/60 cases, including total normalization in 34/60 without lactate concentration modification. A trend toward a decrease in VAT distribution was observed (p =.06). No significant difference was observed in subcutaneous fat distribution, although an increase in subcutaneous thigh adipose tissue was observed in 17/47 (36.2%) cases and 6 patients (12.7%) presented both subcutaneous fat increase and VAT decrease. PMID- 15116285 TI - A phase I study of the pharmacokinetics and safety of passive immunotherapy with caprine anti-HIV antibodies, (PE)HRG214, in HIV-1-infected individuals. AB - PURPOSE: To establish the pharmacokinetics and safety of single-dose polyclonal caprine anti-HIV antibodies ((PE)HRG214)in HIV-1-infected individuals. DESIGN: A phase 1, open-label, nonrandomized, dose-escalating study. METHOD: HIV-1-infected patients with CD4+ T-cell counts of < or =200 cells/microL and plasma HIV viral load (VL)of > or =5,000 copies/mL received a single intravenous dose of HRG. Dosing began at 6,000 U/kg HRG with proposed step-wise escalation to 96,000 U/kg. RESULTS: Eleven males were enrolled; median CD4+T-cell count and VL were 96 cells/microL and 126,200 copies/mL, respectively. HRG exhibited linear pharmacokinetics across the dosing range studied. The mean terminal elimination half-life (t(1/2)) was 136.6 +/- 44.6 hours (range, 52.6-198 h). Serum sickness occurred in one 48,000 U/kg HRG recipient. One 6,000 U/kg and two 24,000 U/kg HRG recipients developed a mild rash. Between baseline and day 60, VL remained unchanged (n = 6), increased by 0.67 log(10) copies/mL (n = 1), or declined by 0.34-1.55 log(10) copies/mL (n = 4). CONCLUSION: Single-dose HRG exhibited linear kinetics and a long half-life. Although numbers in each dosing group were very small (n = 3), HRG was generally well tolerated in doses below 48,000 U/kg. Multiple dosing with HRG in the HIV-salvage setting may be complicated by immune complex formation. PMID- 15116286 TI - Initial therapy for human immunodeficiency virus: broadening the options. AB - The goal of investigation into new therapeutic options for HIV/AIDS is to further the achievements of highly active antiretroviral therapy by developing new drugs with improved efficacy. Although several therapies are currently available for initial therapy in HIV-infected patients, ongoing research focuses on additions to existing and novel drug classes that might have improved pharmacokinetic and tolerability profiles, as well as on new therapeutic combinations that might result in synergistic activity. To retain activity against resistant strains, novel drugs need to target the numerous critical points in the life cycle of HIV, inhibiting different enzyme subsites than those affected by antiretroviral agents currently in use. An improvement in patient adherence to therapy is another key objective of efforts in HIV treatment, as suboptimal drug levels are a main determinant of antiretroviral regimen failure. This article reviews the current classes of antiretroviral agents in development, describing the clinical data obtained to date. These agents may have potential use as initial therapy in HIV patients. PMID- 15116287 TI - Adherence and HIV drug resistance. PMID- 15116289 TI - Refinement of a therapeutic Shiga toxin-binding probiotic for human trials. AB - We have previously constructed a recombinant bacterium expressing a modified lipopolysaccharide (LPS) mimicking the Shiga toxin receptor, which binds toxin with high avidity. This involved cloning Neisseria galactosyl transferase genes (lgtC and lgtE) in pK184 in a derivative of Escherichia coli R1 (CWG308). Such constructs have considerable potential for prevention of disease caused by Shiga toxin-producing E. coli (STEC). However, neither the E. coli host strain nor the expression plasmid is suitable for human use, because the former is derived from a clinical isolate and the latter contains a kanamycin-resistance gene. We have constructed, as a prelude to human trials, a nonpathogenic E. coli K-12 C600 derivative with deletions in waaO and waaB, such that it has the same LPS core structure as CWG308. We also deleted the thyA gene from this strain, rendering it thymine dependent. The kanamycin-resistance gene was also deleted from pK184 and was replaced with Salmonella typhimurium thyA. Neisseria lgtCE was then cloned into this plasmid and transformed into C600 Delta waaOB Delta thyA. The plasmid was stably maintained, and the construct produced a modified LPS and neutralized Stx1 and Stx2c. Moreover, mice challenged with an otherwise fatal dose of STEC were completely protected by oral administration of the novel construct. PMID- 15116290 TI - Ability of blood group A-active glycosphingolipids to act as Escherichia coli heat-labile enterotoxin receptors in HT-29 cells. AB - We examined the ability of blood group A-active glycoconjugates to act as receptors for Escherichia coli heat-labile type I enterotoxin (LT-I) in HT-29 cells. These cells contained ~4 times more specific binding sites for LT-I than for cholera toxin (CT). Binding of LT-I could not be blocked by the B subunit of CT (CT-B), indicating the existence of LT-I receptors in addition to the glycosphingolipid GM1. LT-I was able to increase levels of cyclic adenosine monophosphate (AMP), even in the presence of CT-B. Helix pomatia and anti-blood group A antibody caused a dose-dependent inhibition of binding of LT-I to cells and production of cyclic AMP. LT-I recognized several complex blood group A active glycosphingolipids from cells, and this interaction was also interfered with by H. pomatia. Treatment of cells with D,L-threo-1-phenyl-2 hexadecanoylamino-3-morpholino-1-propanol diminished surface expression of blood group A-active glycosphingolipids and binding of LT-I to non-GM1 receptors. These observations suggest that blood group A-active glycosphingolipids can function as alternative receptors for LT-I in HT-29 cells. PMID- 15116291 TI - Community-onset methicillin-resistant Staphylococcus aureus associated with antibiotic use and the cytotoxin Panton-Valentine leukocidin during a furunculosis outbreak in rural Alaska. AB - BACKGROUND: Community-onset methicillin-resistant Staphylococcus aureus (CO-MRSA) reports are increasing, and infections often involve soft tissue. During a CO MRSA skin infection outbreak in Alaska, we assessed risk factors for disease and whether a virulence factor, Panton-Valentine leukocidin (PVL), could account for the high rates of MRSA skin infection in this region. METHODS: We conducted S. aureus surveillance in the outbreak region and a case-control study in 1 community, comparing 34 case patients with MRSA skin infection with 94 control subjects. An assessment of traditional saunas was performed. S. aureus isolates from regional surveillance were screened for PVL genes by use of polymerase chain reaction, and isolate relatedness was determined by use of pulsed-field gel electrophoresis (PFGE). RESULTS: Case patients received more antibiotic courses during the 12 months before the outbreak than did control subjects (median, 4 vs. 2 courses; P=.01) and were more likely to use MRSA-colonized saunas than were control subjects (44% vs. 13%; age-adjusted odds ratio, 4.6; 95% confidence interval, 1.7-12). The PVL genes were present in 110 (97%) of 113 MRSA isolates, compared with 0 of 81 methicillin-susceptible S. aureus isolates (P<.001). The majority of MRSA isolates were closely related by PFGE. CONCLUSION: Selective antibiotic pressure for drug-resistant strains carrying PVL may have led to the emergence and spread of CO-MRSA in rural Alaska. PMID- 15116292 TI - Reduced adherence and host cell invasion by methicillin-resistant Staphylococcus aureus expressing the surface protein Pls. AB - Pls, the surface protein of methicillin-resistant Staphylococcus aureus (MRSA), prevents adhesion of clinical strain 1061 to immobilized fibronectin (Fn) and immunoglobulin G (IgG). Invasion of mammalian cells by S. aureus depends on Fn mediated binding of staphylococcal Fn-binding proteins to host cell beta (1) integrins. In the present study, we show that, for 10 clinical Pls-positive (Pls(+)) MRSA strains, adhesion to immobilized Fn, fibrinogen (Fg), IgG, and laminin, as well as binding to soluble Fn and Fg, was less efficient than adhesion and binding associated with 4 Pls-negative (Pls(-)) MRSA strains. However, binding to soluble IgG was comparable among both types of strains. For 293 cells, Pls(+) strains were less invasive than were Pls(-) strains (median [range], 35% [22%-70%] and 110% [89%-141%], respectively, compared with strain Cowan 1). Disruption of the pls gene of strain 1061 increased invasiveness, but it did not affect binding of soluble Fn, Fg, and IgG. Complementation restored the low level of invasiveness, but it did not restore the low level of adhesion to immobilized Fn. In conclusion, the reduced adhesiveness and invasiveness of MRSA appear to generally correlate with expression of Pls. PMID- 15116293 TI - Secular trends in hospital-acquired Clostridium difficile disease in the United States, 1987-2001. AB - We reviewed Clostridium difficile-associated disease (CDAD) data from the intensive care unit (ICU) and hospital-wide surveillance components of the National Nosocomial Infections Surveillance System hospitals during 1987-2001. ICU CDAD rates increased significantly only in hospitals with >500 beds (P<.01) and correlated with the duration of ICU stay (r=0.82; P<.05). Hospital-wide (non ICU) rates increased only in hospitals with <250 beds (P<.01) and in general medicine patients versus surgery patients (P<.0001). CDAD predominated in general hospitals versus other facility types, and rates were significantly higher during winter versus nonwinter months (P<.01). Thus, prevention efforts should be targeted to high-risk groups in these settings. PMID- 15116294 TI - Relationship between fluoroquinolone area under the curve: minimum inhibitory concentration ratio and the probability of eradication of the infecting pathogen, in patients with nosocomial pneumonia. AB - Our objective was to prospectively determine the factors influencing the probability of a good microbiological or clinical outcome in patients with nosocomial pneumonia treated with a fluoroquinolone. Levofloxacin was administered as an infusion of 500 mg/h for 1.5 h (total dose, 750 mg). For patients with Pseudomonas aeruginosa or methicillin-resistant Staphylococcus aureus, a second drug was added (ceftazidime or piperacillin/tazobactam for P. aeruginosa and vancomycin for methicillin-resistant S. aureus). Population pharmacokinetic studies of 58 patients demonstrated that this population handled the drug differently from populations of volunteers. Multivariate logistic regression analysis (n=47 patients) demonstrated that only the age of the patient and the achievement of an area under the curve: minimum inhibitory concentration ratio of > or =87 had a significant effect on eradication of the pathogen (P<.001). Achieving the breakpoint made the patient 4 times more likely to achieve eradication. The effect was greatest in patients > or =67 years old. PMID- 15116295 TI - Dominance of CD86, transforming growth factor- beta 1, and interleukin-10 in Mycobacterium tuberculosis secretory antigen-activated dendritic cells regulates T helper 1 responses to mycobacterial antigens. AB - We report that stimulation of Mycobacterium tuberculosis (M. tuberculosis) secretory antigen (MTSA)-differentiated dendritic cells (DCs) and MTSA-matured DCs with M. tuberculosis cell extract (CE) down-regulated proinflammatory responses to CE-primed T (CE-T) cells by increasing surface expression of CD86 after CE stimulation. CE stimulation also decreased interleukin (IL)-12p40 and interferon (IFN)- gamma levels and increased IL-10 and transforming growth factor beta 1 (TGF- beta 1) levels from these DCs. Blocking either CD86, IL-10, or TGF- beta with monoclonal antibodies before CE stimulation restored the attenuated T helper 1 (Th1) responses of CE-T cells. Conversely, treatment of these DCs with IL-12p70 and/or IFN- gamma completely restored Th1 responses from CE-T cells. These results indicate that M. tuberculosis secretory antigens down-regulate proinflammatory Th1 responses to mycobacteria by differentially modulating the cytokine profiles and surface densities of costimulatory molecules on DCs. Of importance, this down-regulation is independent of the maturation status of MTSA activated DCs and can be rescued after treatment of DCs with IFN- gamma or IL-12. PMID- 15116296 TI - Phagosomal acidification is not a prerequisite for intracellular multiplication of Legionella pneumophila in human monocytes. AB - Legionella pneumophila is able to multiply in a variety of eukaryotic cells. Contradictory results have been published on the significance of phagosomal acidification in the intracellular multiplication of Legionella species in monocytes. Therefore, we analyzed the phagosomal pH values in 2 different types of human monocytes throughout the intracellular-replication cycles of 2 Legionella species that have different rates of intracellular multiplication. Our results show that phagosomal acidification is not a prerequisite for intracellular replication of Legionella pneumophila in human monocytes. PMID- 15116297 TI - Absence of cytomegalovirus-resistance mutations after valganciclovir prophylaxis, in a prospective multicenter study of solid-organ transplant recipients. AB - We investigated the emergence of cytomegalovirus (CMV) ganciclovir-resistance mutations in 301 high-risk solid-organ transplant (SOT) recipients after oral prophylaxis, for 100 days, with either valganciclovir or ganciclovir. For patients treated with ganciclovir, the incidence of CMV UL97 mutations was 1.9% (2/103) at the end of prophylaxis and 6.1% (2/33) for patients with suspected CMV disease up to 1 year after transplantation. No resistance mutations were detected in samples from valganciclovir-treated patients. Dual polymerase (UL54) and UL97 resistance mutations were not seen. Valganciclovir was associated with negligible risk of resistance and thus constitutes a useful alternative to ganciclovir prophylaxis for CMV in high-risk SOT recipients. PMID- 15116298 TI - Antiviral CD8 T cells in the control of primary human cytomegalovirus infection in early childhood. AB - Human cytomegalovirus (CMV) establishes persistent infection, with control of replication thought to be mediated by CMV-specific CD8 T cells. Primary CMV infection commonly affects young children and causes prolonged viral shedding in saliva and urine. We investigated whether this virus-host interaction pattern reflects a developmental deficiency of antiviral CD8 T cell-mediated immunity during childhood. CMV-specific CD8 T cell responses in asymptomatic children with active infection were not different from adults with recent or long-term infection in frequency and functional analyses. High urine CMV concentrations were detected, despite these CMV-specific CD8 T cell responses. We conclude that delayed resolution of primary CMV infection in young children is not caused by a deficient CMV-specific CD8 T cell response. Because these healthy children continue to have local CMV replication, we suggest that CD8 T cells may function primarily to prevent symptomatic, disseminated disease. PMID- 15116299 TI - Activation of porcine cytomegalovirus, but not porcine lymphotropic herpesvirus, in pig-to-baboon xenotransplantation. AB - Tissue-invasive disease due to porcine cytomegalovirus (PCMV) has been demonstrated after pig-to-baboon solid-organ xenotransplantation. Porcine lymphotropic herpesvirus (PLHV)-1 is associated with B cell proliferation and posttransplant lymphoproliferative disorder after allogeneic bone marrow transplantation in swine but has not been observed in pig-to-primate xenotransplantation. Activation of PCMV and PLHV-1 was investigated in 22 pig-to baboon xenotransplants by use of quantitative polymerase chain reaction. PCMV was found in all xenografts; increased viral replication occurred in 68% of xenografts during immunosuppression. PLHV-1 was found in 12 xenografts (55%); no increases in viral replication occurred during immunosuppression. Control immunosuppressed swine coinfected with PCMV and PLHV-1 had activation of PCMV but not PLHV-1. PCMV, but not PLHV-1, is activated in solid-organ xenotransplantation. PMID- 15116300 TI - Intracellular distribution of hepatitis B virus core protein expressed in vitro depends on the sequence of the isolate and the serologic pattern. AB - Intracellular localization of hepatitis B core antigen (HBcAg) in vivo varies with liver cell damage. Localization of HBcAg was studied using transfection of cloned HBcAg variants. Twenty-six samples were obtained from 14 patients with liver disease; 10 were hepatitis B e antigen positive, and 16 were anti-hepatitis B e (HBe) positive. In hepatitis B e antigen (HBeAg)-positive patients, HBcAg predominantly localized in the nucleus; in anti-HBe-positive patients, it accumulated mainly in the cytoplasm. Of the 13 samples with nuclear localization, 9 were HBeAg positive; 5 of 13 had C-terminus and/or B cell epitope mutations. All but 1 of the 13 samples with predominantly cytoplasmic localization were anti HBe positive; all 13 had mutations. Reversion of mutant sequences with cytoplasmic expression back to the wild type led to a shifting back to nuclear distribution. Thus, the pattern of HBcAg localization in vitro depends on sequence and the serologic pattern of chronic infection, paralleling the situation in vivo. PMID- 15116301 TI - Quantification and functional analysis of plasmacytoid dendritic cells in patients with chronic hepatitis C virus infection. AB - BACKGROUND: Plasmacytoid dendritic cells (PDCs) are the major producers of interferon (IFN)- alpha within peripheral blood mononuclear cells (PBMCs). METHODS: We analyzed whether chronic hepatitis C virus (HCV) infection could be linked to a defective function or number of PDCs. We evaluated the capacity of PBMCs from 5 cohorts of subjects to produce IFN- alpha after viral stimulation. We concomitantly analyzed the frequency of PDCs and the levels of IFN- alpha transcripts within the PBMCs from the same cohorts. RESULTS: PBMCs from patients with chronic HCV infection receiving antiviral therapy displayed a reduced capacity to release IFN- alpha, compared with those from healthy individuals, those from long-term responders to therapy, and those from nontreated patients. This defect was significantly correlated with the percentage of PDCs. In addition, PDCs from patients with chronic HCV infection receiving therapy displayed a reduced intrinsic capacity to produce IFN- alpha, which could be linked to the level of IFN- alpha transcripts. CONCLUSION: Our observations point to an effect of the therapy on either the survival or the localization of PDCs, rather than a direct detrimental effect due to the viral infection during chronic HCV infection. PMID- 15116302 TI - Effect of Epstein-Barr virus replication on Langerhans cells in pathogenesis of oral hairy leukoplakia. AB - Epstein-Barr virus (EBV) replicates productively in oral hairy leukoplakia (HLP). One characteristic of human immunodeficiency virus (HIV)-associated HLP is a decreased oral epithelial Langerhans cell count. This prospective study tested the hypothesis that oral epithelial EBV replication decreases oral Langerhans cell counts. EBV replication in HLP was highly correlated with decreased oral Langerhans cell counts. Inhibition of EBV replication restored oral Langerhans cell counts to normal control levels, and the return of EBV replication after treatment resulted in a recurrent decline in oral Langerhans cell counts. Decreased oral Langerhans cell counts occurred independently of HIV infection, as demonstrated in HLP of otherwise healthy HIV-seronegative individuals. These results support the tested hypothesis and suggest that EBV manipulates and evades the mucosal immune response in oral epithelial infection. This novel EBV strategy for eliminating oral Langerhans cells may facilitate the persistence of oral epithelial EBV and may contribute to the pathogenesis of HLP. PMID- 15116303 TI - Isolation and molecular characterization of a novel type 3 reovirus from a child with meningitis. AB - Mammalian reoviruses are non-enveloped viruses that contain a segmented, double stranded RNA genome. Reoviruses infect most mammalian species, although infection with these viruses in humans is usually asymptomatic. We report the isolation of a novel reovirus strain from a 6.5-week-old child with meningitis. Hemagglutination and neutralization assays indicated that the isolate is a serotype 3 strain, leading to the designation T3/Human/Colorado/1996 (T3C/96). Sequence analysis of the T3C/96 S1 gene segment, which encodes the viral attachment protein, sigma 1, confirmed the serotype assignment for this strain and indicated that T3C/96 is a novel reovirus isolate. T3C/96 is capable of systemic spread in newborn mice after peroral inoculation and produces lethal encephalitis. These results suggest that serotype 3 reoviruses can cause meningitis in humans. PMID- 15116304 TI - Monophyletic relationship between severe acute respiratory syndrome coronavirus and group 2 coronaviruses. AB - Although primary genomic analysis has revealed that severe acute respiratory syndrome coronavirus (SARS CoV) is a new type of coronavirus, the different protein trees published in previous reports have provided no conclusive evidence indicating the phylogenetic position of SARS CoV. To clarify the phylogenetic relationship between SARS CoV and other coronaviruses, we compiled a large data set composed of 7 concatenated protein sequences and performed comprehensive analyses, using the maximum-likelihood, Bayesian-inference, and maximum-parsimony methods. All resulting phylogenetic trees displayed an identical topology and supported the hypothesis that the relationship between SARS CoV and group 2 CoVs is monophyletic. Relationships among all major groups were well resolved and were supported by all statistical analyses. PMID- 15116305 TI - A prospective, blinded study of quantitative touch-down polymerase chain reaction using oral-wash samples for diagnosis of Pneumocystis pneumonia in HIV-infected patients. AB - Oral-wash samples obtained during 113 episodes of suspected Pneumocystis pneumonia (PCP) in human immunodeficiency virus-infected patients were tested by use of a quantitative touch-down PCR (QTD PCR) assay. QTD PCR had a sensitivity of 88% and a specificity of 85%. Treatment for PCP prior to oral wash collection had an impact on the sensitivity, and PCR-positive oral-wash samples obtained within < or =1 day of treatment from patients without PCP had significantly fewer copies per tube than did those from patients with PCP; thus, application of a post hoc cut-off value of 50 copies/tube increased the specificity to 100%. QTD PCR of oral-wash samples can be an accurate and noninvasive method for diagnosis of PCP. PMID- 15116306 TI - Increase in prevalence of Pneumocystis carinii mutations in patients with AIDS and P. carinii pneumonia, in the United States and China. AB - This study of Pneumocystis carinii dihydropteroate synthase (DHPS) mutations in patients with AIDS who have P. carinii pneumonia compares the change in the prevalence of such mutations in the United States, where sulfa-drug prophylaxis is widespread, to that in China, where it is infrequent. The DHPS gene from 145 US patients presenting during 1983-2001 and from 15 Chinese patients presenting during 1998-2001 was amplified by polymerase chain reaction and was sequenced. In the United States, 40% of patients had DHPS mutations; 38% received sulfa-drug prophylaxis. Mutation prevalence increased to 70% during 2000-2001, from 25% during 1994-1995 (P<.01). In China, 7% of patients had DHPS mutations; none received sulfa-drug prophylaxis. The prevalence of P. carinii DHPS mutations has markedly increased in the United States but remains low in China. PMID- 15116307 TI - Mutations in HIV-1 reverse transcriptase potentially associated with hypersusceptibility to nonnucleoside reverse-transcriptase inhibitors: effect on response to efavirenz-based therapy in an urban observational cohort. AB - BACKGROUND: Hypersusceptibility to nonnucleoside reverse-transcriptase inhibitors (NNRTIs) was described in association with reverse-transcriptase (RT) mutations conferring resistance to nucleoside reverse-transcriptase inhibitors (NRTIs). We evaluated the effect of RT mutations associated with hypersusceptibility to NNRTIs on the response to efavirenz-based therapy. METHODS: We analyzed an observational database of patients for whom highly active antiretroviral therapy failed and who received genotypic resistance testing-guided therapy, either efavirenz or protease inhibitor (PI) based. Study end points were achievement of virus load <80 copies/mL, achievement of virus load <80 copies/mL without rebound to >500 copies/mL, and changes in CD4 cell counts. RESULTS: The baseline RT mutations M41L, M184V, L210W, and T215Y and the M41L/T215Y and M41L/T215Y/M184V combinations were associated with virological suppression for efavirenz-treated patients, whereas, for PI-treated patients, only the M184V mutation was associated with virological suppression, and the L210W mutation showed a negative correlation; no correlation was found between any mutation and virological response without rebound. CONCLUSIONS: The M41L, M184V, L210W, and T215Y mutations were associated with a better, although transient, virological outcome in patients treated with efavirenz-based regimens. PMID- 15116308 TI - HLA-DRB1 and -DQB1 alleles and haplotypes in Zambian couples and their associations with heterosexual transmission of HIV type 1. AB - In 292 initially human immunodeficiency virus (HIV)-1-serodiscordant and cohabiting Zambian couples, HLA-DRB1 and -DQB1 variants were associated with HIV 1 transmission events during a 7-year follow-up period. Initially seronegative partners with either DRB1*0301-DQB1*0201 (relative hazard [RH], 1.60; P=.009) or DRB1*1503-DQB1*0602 (RH, 1.67; P=.03) showed accelerated seroconversion. Carriage of DRB1*1301 in initially seropositive partners led to delayed transmission of HIV to their spouses (RH, 0.54; P=.05). The combined groups of seroprevalent and seroincident partners (n=433) also differed from those who remained seronegative (n=151), with regard to 2 common haplotypes, DRB1*1302-DQB1*0604 (relative odds [RO], 0.28; P=.003) and DRB1*1503-DQB1*0602 (RO, 1.81; P=.02). Statistical adjustments for other host factors (age, sex, genital ulcer, and index partner's virus load) known to influence transmission of HIV-1 seldom altered the genetic relationships. Overall, associations of HLA class II polymorphisms with both HIV transmission and acquisition are not as readily interpretable as are effects reported for other loci. PMID- 15116309 TI - Distinct patterns of peripheral HIV-1-specific interferon- gamma responses in exposed HIV-1-seronegative individuals. AB - It is unclear how human immunodeficiency virus (HIV) type 1-specific immune responses in exposed seronegative (ESN) individuals differ from those in HIV-1 infected subjects. By use of overlapping peptides spanning Gag, Tat, Nef, Vif, Vpr, and Vpu, peripheral blood mononuclear cells from ESN individuals, their seropositive (SP) partners, and unexposed seronegative control subjects were screened for interferon- gamma production. Responses were more frequent (95.7% vs. 20%), of a higher magnitude (9-fold), and of wider breadth (median number of peptides recognized, 18 vs. 2.5) in SP than in ESN individuals. Peptides recognized by ESN individuals were less frequently recognized by their SP partners. SP subjects infrequently recognized peptides from Vif, and such responses were subdominant; among ESN individuals, this HIV-1 protein was most frequently recognized. Immunodominant peptides recognized by SP subjects tended to be from relatively conserved regions, whereas peptides recognized by ESN individuals were associated with slow disease progression. PMID- 15116310 TI - Alterations in expression of monocyte chemotactic protein-1 in the simian immunodeficiency virus model of disseminated Mycobacterium avium complex. AB - Mycobacterium avium complex (MAC) infection is the most common disseminated bacterial infection in untreated patients with acquired immunodeficiency syndrome (AIDS). We investigated the potential role of monocyte chemotactic protein-1 (MCP 1) in the pathogenesis of disseminated MAC, using the simian immunodeficiency virus (SIV)/macaque model of AIDS. Macaques were inoculated with SIV, followed by challenge with a pathogenic AIDS isolate of M. avium 14 days later. After challenge with M. avium, marked increases in serum MCP-1 levels were detected in SIV-infected macaques, a finding that was duplicated in coinoculated bronchoalveolar macrophages. MCP-1 levels were significantly higher in SIV infected macaques than in non-SIV-infected controls (327.1 vs. 151.5 pg/mL, respectively; P=.04), suggesting that up-regulation of MCP-1 contributes to the development of progressive mycobacterial disease. Similarly, morphometric analysis revealed increased expression of MCP-1 in hepatic microgranulomas from SIV-infected macaques. We conclude that the pronounced increases in MCP-1 levels demonstrated in tissue and serum samples after M. avium inoculation may play a role in the development of disseminated mycobacterial disease. PMID- 15116311 TI - Age-associated decline in resistance to Babesia microti is genetically determined. AB - BACKGROUND: Although infection by the protozoan Babesia microti is rarely symptomatic in immunocompetent young people, healthy individuals aged >50 years may experience life-threatening disease. To determine the basis for this age relationship, we developed a mouse model of babesiosis using a novel clinical isolate of B. microti. METHODS: Mice were infected at 2, 6, 12, or 18 months. Parasitemia was monitored on Giemsa-stained blood smears or by flow cytometry. RESULTS: In DBA/2 mice, early and persistent parasitemias increased with age at infection. BALB/c and C57BL/6 mice were resistant, regardless of age, which indicates that allelic variation determines resistance to B. microti. Unlike immunocompetent mice, SCID mice, which retain an innate immune system but lack the lymphocytes needed for adaptive immunity, developed high and persistent levels of parasitemia that were markedly reduced by transfer of naive BALB/c or DBA/2 splenocytes. BALB/c cells reduced the persistent parasitemia to a greater extent than did age-matched DBA/2 cells. Of importance, there was an age associated loss of protection by cells of both strains. CONCLUSION: The resistance to B. microti infection conferred by the adaptive immune system is genetically determined and associated with age. We postulate that there are age related differences in the expression of alleles critical for adaptive immunity to B. microti. PMID- 15116312 TI - Polymerase chain reaction using noninvasively obtained samples, for the detection of Leishmania infantum DNA in dogs. AB - A polymerase chain reaction (PCR) procedure using noninvasively obtained samples, for the identification of Leishmania infantum in canine tissues, was evaluated and compared with serologic testing and culture. A total of 92% of naturally infected, symptomatic, seropositive dogs were found to be positive by use of DNA from conjunctival swabs. Spleen or lymph node aspirates were found to be positive by PCR in 86% and by culture in 74% of these dogs. The sensitivity and specificity of conjunctival PCR were 92% and 100%, respectively. Experimentally infected dogs were found to be positive by conjunctival PCR already at 45 days of infection (83%) and before seroconversion. PCR using noninvasively obtained conjunctival samples will be useful for epidemiological studies and for direct diagnosis of canine visceral leishmaniasis. PMID- 15116313 TI - PapG-dependent adherence breaks mucosal inertia and triggers the innate host response. AB - Mucosal pathogens differ from normal flora constituents in that they provoke a host response that upsets mucosal integrity. We investigated whether the elaboration of discrete adherence factors is sufficient to break the inertia of the mucosal barrier. PapG-mediated adherence was selected as an example, because P fimbrial expression characterizes uropathogenic Escherichia coli and because adherence starts the attack on the mucosal barrier. Patients were inoculated intravesically with transformed nonvirulent E. coli strains expressing functional P fimbriae (E. coli pap(+)) or mutant fimbriae lacking the adhesin (E. coli Delta papG). E. coli pap(+) was shown to activate the innate host response, and adherent gfp(+) bacteria were observed on excreted uroepithelial cells. E. coli Delta papG failed to trigger a response and was nonadhesive. We conclude that PapG-mediated adherence breaks mucosal inertia in the human urinary tract by triggering innate immunity and propose that this activation step differentiates asymptomatic carriage from infection. PMID- 15116314 TI - Is anti-tumor necrosis factor therapy associated with increased mortality in patients with severe sepsis caused by pneumonia? PMID- 15116315 TI - Experimental investigations regarding the etiology of dengue fever. 1907. PMID- 15116316 TI - Offspring gender ratio and the rate of recurrent spontaneous miscarriages in jewish women at high risk for breast/ovarian cancer. AB - BRCA1/BRCA2 germline mutations are associated with an increased breast/ovarian cancer risk. Offspring gender ratios may be skewed against male births in BRCA1 mutation carriers. In addition, the lack of viable homozygous BRCA1/BRCA2 mutation carriers implies that recurrent miscarriages may be associated with homozygous fetuses. Jewish Israeli high-risk women who were tested for being carriers of the predominant BRCA1/BRCA2 mutations in Jewish high-risk families were analyzed for the sex of offspring and the rate of spontaneous miscarriages. Overall, 817 women participated: 393 BRCA1/BRCA2-mutation carriers (229 with breast/ovarian cancer) and 424 high-risk noncarriers (208 with breast/ovarian cancer). No differences between the male-to-female offspring ratios of all study groups were noted. Among mutation carriers, the offspring male-to-female ratio was 0.97 (444 : 460), and among mutation carriers with cancer it was 0.92 (262 : 284). Similarly, no offspring gender skewing was noted among high-risk noncarriers, regardless of health status. The rates of three or more spontaneous miscarriages among participants with at least one live birth were 4.37% (15/343) among mutation carriers and 3% (12/401) among high-risk women (P = not significant). In conclusion, the offspring gender ratio is similar in high-risk Jewish families and in the general population. The issue of the rate of recurrent miscarriages in high-risk Jewish women is unresolved. PMID- 15116317 TI - Telomerase: promise and challenge. PMID- 15116318 TI - Telomerase and human papillomavirus as diagnostic adjuncts for cervical dysplasia and carcinoma. AB - Telomerase and human papillomavirus (HPV) DNA were evaluated as potential markers of high-grade dysplasia in cervical cytological specimens. Cytology specimens were collected from patients at the time of colposcopic evaluation for management of a previous abnormal cytology test result. Telomerase activity was evaluated by the telomeric repeat amplification protocol (TRAP), and HPV DNA was detected by polymerase chain reaction with L1 consensus-sequence primers and filter hybridization genotyping. Telomerase was detected in 8 of 97 (8.2%) cases with normal cytology or benign cellular changes, in 7 of 98 (7.1%) cases of atypical squamous cells of undetermined significance (ASCUS), in 3 of 95 (3.2%) cases of low-grade squamous intraepithelial lesion (LSIL), and in 17 of 48 (35.4%) cases with high-grade squamous intraepithelial lesion (HSIL). High-risk HPVs were detected in 23 of 97 (23.7%) cases with normal/reactive cellular changes (RCC) cytology, in 28 of 98 (28.6%) cases of ASCUS, in 69 of 95 (72.6%) cases of LSIL, and in 35 of 48 (72.9%) cases of HSIL. Telomerase expression did not correlate with the detection of high-risk HPVs in any cytological diagnostic categories. Telomerase and HPV test results of cytological specimens were correlated with the histological diagnoses of concurrent cervical biopsy specimens. Telomerase showed a sensitivity of 29.9% and a specificity of 94.0% for biopsy-confirmed cervical intraepithelial neoplasia (CIN) II/III. In contrast, high-risk HPVs were detected in 70.1% of cases with underlying CIN II/III, with a specificity of 62.5%. A relatively high proportion of normal/RCC or ASCUS cases with telomerase-positive test results had underlying high-grade dysplasia on cervical biopsy. Thus, technical and practical limitations of the TRAP assay in cervical cytology specimens limit the practical application of telomerase as a diagnostic adjunct in cervical cytopathology. PMID- 15116320 TI - Myocytes of chorionic vessels from placentas with meconium-associated vascular necrosis exhibit apoptotic markers. AB - Meconium-associated vascular necrosis (MAVN) is a histological abnormality of human placental chorionic vessels that is associated with poor neonatal outcome. We tested the hypothesis that MAVN shows apoptosis in the walls of chorionic vessels. Archival placental specimens with MAVN (n = 5) were compared with specimens from uncomplicated pregnancies at term (n = 5) and from placentas with intense chorionic vasculitis associated with acute chorioamnionitis with (n = 5) or without (n = 5) a clinical history of meconium in the amniotic fluid. Sections from all placentas were processed by the TUNEL method, and 2 observers who were blinded to specimen diagnosis quantified the immunofluorescent TUNEL staining in both the amnion-facing and villous-facing walls of the larger chorionic vessels in each specimen. Compared with the other 3 groups, only the amnion-facing wall of chorionic vessels in MAVN showed a significantly greater number of apoptotic cells. This was verified by morphological criteria and caspase 3 staining. There were limited or no detectable TUNEL-stained cells in either the villous-facing walls of vessels in the MAVN specimens or in any of the vessels of the placentas from uncomplicated pregnancies. There was a negligible level of apoptosis in chorionic vessels of placentas with intense chorionic vasculitis, with or without meconium, despite the inflammatory response or presence of meconium. We conclude that apoptosis contributes to the pathophysiology of MAVN. PMID- 15116319 TI - Telomere length, telomerase activity, and expression of human telomerase reverse transcriptase mRNA in growth plate of epiphyseal articular cartilage in femoral head during normal human development and in thanatophoric dysplasia. AB - Telomeres are important in chromosome structure and function, protecting against their degradation. However, few studies have examined telomeres in growth plates within articular cartilage during normal development. We investigated frozen sections that were obtained from 57 reference autopsy cases (aged from 16 weeks of gestation to 91 years) and from 2 patients with thanatophoric dysplasia. In the reference cases, telomere length was significantly longer in growth plates obtained from the 10 cases that were aged from 16 weeks of gestation to 10 years than in those from 47 of the adult cases (aged 20 to 91 years). In fetal, neonatal, and child cases, telomerase activity was significantly higher in the hypertrophied zone (HZ) in growth plates than in the other 3 zones. The hTERT mRNA staining intensity (staining area) was stronger (larger) in HZ and the proliferating zone than in the calcified zone and resting zone. In thanatophoric dysplasia, telomere length and telomerase activity were short and low, respectively, compared with those of normal growth plates at an equivalent age, and expression of hTERT mRNA was negative or weakly positive in all 4 zones within growth plates. These results suggest that telomere length and telomerase activity have significant effects in the growth plates of articular cartilage, particularly at developmental ages from fetus to child. We speculate that short telomere length and low telomerase activity may be important for chondrocyte differentiation in rhizomeric shortening of the limbs in thanatophoric dysplasia. PMID- 15116321 TI - Loss of heterozygosity in clonal evolution with genetic progression and divergence in spindle cell carcinoma of the gallbladder. AB - Spindle cell carcinoma (SpCC) of the gallbladder is a rare neoplasm that shows carcinoma with a variable component of sarcomatoid spindle cells. The clinical and pathological features of this neoplasm have been well documented, but the histogenesis has long been a matter of speculation. In an attempt to clarify the clonality and genetic relationships involved in the evolution of this neoplasm, we microdissected a total of 18 carcinomatous and sarcomatous foci from 2 gallbladder SpCCs and analyzed the allelic status with 42 microsatellite markers on chromosomal arms 1p, 1q, 3p, 4q, 5q, 6q, 8p, 9p, 10q, 11p, 11q, 13q, 16q, 17p, 17q, 18q, and 22q. The 2 cases examined had a polypoid tumor in the gallbladder, in which both adenocarcinomatous and sarcomatoid spindle cell components were identified histologically. In both SpCCs, homogenous allelic losses were identified in both the carcinomatous and sarcomatous components; 17p, 18q, and 5q in case 1 and 17p and 11q in case 2. These indicated that both SpCCs had a single clonal origin. In case 1, additional loss of heterozygosity (LOH; 6q) consisting of genetic progression occurred in both the carcinomatous and sarcomatoid components. In case 2, there was additional LOH (9p) in the carcinomatous components and additional microsatellite instability at D5S644 in both the carcinomatous and sarcomatoid components, indicating a monoclonal neoplasm with genetic progression and divergence. In the 2 cases, the genetic changes indicated that an original clone of a pure adenocarcinoma apparently acquired sarcomatoid spindle cell phenotype by successive genetic changes. On the other hand, we saw no evidence of tumors in which a sarcomatoid spindle cell appeared to give rise to a carcinomatous subclone in the examined cases. In conclusion, the current study includes the first LOH analyses of SpCC of the gallbladder. Our data support the concept that gallbladder SpCC is derived from a single clone originating from a carcinoma. Furthermore, we showed genetic heterogeneity accompanying the phenotypic divergence, with patterns of genetic alterations that are consistent with both the progression and divergence within the individual tumors. PMID- 15116322 TI - Overexpression of metastatic tumor antigen 1 in hepatocellular carcinoma: Relationship to vascular invasion and estrogen receptor-alpha. AB - The morbidity and mortality experienced by cancer patients is mainly due to the invasion and metastasis of the primary tumor. Recently, a potential metastasis associated gene and its product, the metastatic tumor antigen 1 (MTA1), were identified; this gene has been found to be overexpressed in a variety of cancers. MTA1 is also known as a potent co-repressor of estrogen receptor element transcription in breast cancer cells. The expression of MTA1 in hepatocellular carcinoma (HCC) and its potential relationship to metastasis and to estrogen receptor alpha (ER-alpha) expression has not been examined, forming the basis for this study. Paraffin sections of 45 HCC specimens, 4 different HCC cell lines, and normal hepatocyte cell line (h NHeps) were immunostained with MTA1 and ER alpha antibodies. In addition, we examined, by Western blotting, the MTA1 and ER alpha expression levels in 4 human HCC lines (HepG2 [wild p53], HLE, HLF, and HuH 7 [mutant p53]). MTA1 was overexpressed in HCC cells versus nonmalignant hepatocytes in 31 of 45 HCC specimens (69%). Its expression was predominantly localized to the nucleus or cytoplasm of HCC cells. Nineteen of 20 HCC (95%) specimens with vascular invasion displayed strong MTA1 expression. Overexpression of MTA1 also significantly correlated with large tumor size. The cytoplasmic and nuclear immunoreactivity for ER-alpha was present in HCC specimens in 46% and 12%, respectively. Expression of MTA1 inversely correlated with the nuclear localization of ER-alpha. There was no marked difference in MTA1 and ER-alpha expression levels between HCC cell line expressing wild-type p53 and cell line with mutated p53 HCC. In conclusion, these findings indicate that overexpression of MTA1 is associated with HCC growth and vascular invasion. Nuclear translocation of ER-alpha inversely correlated with MTA1 expression, suggesting negative regulatory mechanisms. PMID- 15116323 TI - Assessment of donor liver steatosis: pathologist or automated software? AB - Steatosis in donor liver biopsy specimens has been shown to correlate with graft dysfunction after orthotopic liver transplantation. This 2-part (laboratory pilot, clinical retrospective) study compared the traditional interpretation of steatosis by a pathologist with an automated measurement determined by an image analysis system. In our pilot study, Sprague-Dawley rats were studied prospectively by feeding them a choline-deficient diet for up to 7 days. In our clinical group, data from 49 consecutive recipients of cadaveric liver transplantation were reviewed retrospectively. In both studies, the percentages of microvesicular fat, macrovesicular fat, and total fat content within liver biopsy specimens were determined by an automated image analysis software program and a pathologist using the same set of slides. The association between fat content of the donor liver and patient survival and graft survival, along with levels of aspartate aminotransferase, alanine aminotransferase, prothrombin time, and total bilirubin after transplantation, were also examined in the clinical study. A direct correlation was observed between levels of macrovesicular fat determined by a pathologist and the automated software using livers from rats fed a choline-deficient diet and livers from deceased donors. A significant association was observed between macrovesicular fat content in the donor liver biopsy and graft survival by both techniques. We conclude that an image analysis system can be used to automate the determination of fat content in liver biopsy specimens, and that its findings correlate with both the visual interpretation by a pathologist and graft survival. Further study is needed to determine the role of an automated technique in the evaluation of donor livers for transplantation. PMID- 15116324 TI - Ultrastructural localization of thyroid peroxidase, hydrogen peroxide-generating sites, and monoamine oxidase in benign and malignant thyroid diseases. AB - Despite thyroid tissue heterogeneity, biochemical and morphological features have been associated with certain thyroid diseases. We analyzed the ultracytochemical localization of thyroperoxidase (TPO), TPO-associated hydrogen peroxide generating sites (H(2)O(2) sites), and monoamine oxidase (MAO) in terms of morphology and biochemical TPO activity in abnormal thyroids. We examined 11 cases of nontoxic multinodular goiter, 5 cases of Hashimoto's thyroiditis, 1 case of oncocytic (Hurthle or oxyphilic cell) adenoma, 5 cases of Graves' disease, 4 cases of papillary carcinoma, and 4 cases of perinodular normal tissue. In the perinodular tissue, TPO was detected mainly in the nuclear envelope, rough endoplasmic reticulum (RER), and subapical vesicles, but not in the apical surface. In multinodular goiter, heterogeneous TPO reactivity ranging from almost null to strongly positive was detected in similar locations as in the perinodular tissue, and was absent in the microvilli. Follicular cells from Hashimoto's thyroiditis displayed TPO in the nuclear envelope and the scarce RER. Remarkably, oncocytic cells from both Hashimoto's thyroiditis and oncocytic adenoma, typically packed with mitochondria, displayed evident TPO reaction exclusively in mitochondrial cristae. In Graves' disease, the nuclear envelope, enlarged RER, and apical vesicles were strongly TPO positive, and microvilli also exhibited TPO activity. Papillary carcinoma cells were negative for TPO. The localization and characteristics of TPO activity in the H(2)O(2) sites were similar to that of TPO in all tissues. MAO was positive in mitochondria of perinodular tissues, multinodular goiter, and oncocytes and negative in Hashimoto's thyroiditis and Graves' disease. Interestingly, MAO was intensely positive in the nuclear envelope of papillary carcinoma but unreactive in mitochondria. Biochemical TPO activity was increased in multinodular goiter and Graves' disease. In conclusion, several changes in ultracytochemical characteristics of TPO, H(2)O(2) sites, and MAO were associated with thyroid disease. Nonmalignant oncocytic cells exhibited an unusual mitochondrial location of TPO and H(2)O(2) sites. The distribution of MAO in nuclear envelope of papillary carcinoma cells could be a further feature of malignancy. PMID- 15116325 TI - Lack of PAX5 rearrangements in lymphoplasmacytic lymphomas: reassessing the reported association with t(9;14). AB - A t(9;14)(p13;q32) involving the PAX5 and IGH genes has been described in association with lymphoplasmacytic lymphoma. Although often described as common, the incidence of this translocation in nodal lymphoplasmacytic lymphoma has never been investigated. Recent studies of patients with Waldenstrom's macroglobulinemia (often corresponding to marrow-based lymphoplasmacytic lymphoma) have failed to identify the t(9;14). These studies have suggested that either nodal and marrow-based lymphoplasmacytic lymphomas have distinct pathogenetic mechanisms or that the t(9;14) is less frequent in lymphoplasmacytic lymphoma than was believed previously. We therefore analyzed a series of nodal or other extramedullary lymphoplasmacytic lymphomas for the presence of the t(9;14) with paraffin section interphase fluorescence in situ hybridization. We developed a BAC contig probe spanning all previously described PAX5 breakpoints and validated this assay with the KIS-1 cell line that expresses a t(9;14). Analysis with the PAX5 probe showed a lack of PAX5 rearrangements in all cases that were analyzed successfully. Similarly, analysis by an IGH fluorescence in situ hybridization probe showed no evidence of translocations involving the IGH locus. These findings indicate that the t(9;14) is at least uncommon in lymphoplasmacytic lymphoma and should no longer be considered a characteristic finding in this type of lymphoma as defined by World Health Organization criteria. PMID- 15116326 TI - Absence of Epstein-Barr virus in anaplastic large cell lymphoma: a study of 64 cases classified according to World Health Organization criteria. AB - The frequency of Epstein-Barr virus (EBV) in anaplastic large cell lymphoma (ALCL) has been controversial. The interpretation of previous studies is complicated by the use of nonuniform EBV detection methods and the inclusion of cases of CD30-positive diffuse large B-cell lymphoma and so-called "ALCL, Hodgkin like," as defined in the Revised European-American Lymphoma classification scheme. In the current World Health Organization (WHO) classification system, both of these tumors are excluded from the ALCL category. Also, recently developed antibodies (eg, the antibody specific for PAX-5/B-cell-specific activator protein [BSAP]) provide new, sensitive tools for identifying neoplasms of B-cell lineage that can morphologically resemble ALCL. In this study we evaluated 64 cases of ALCL of T- or null-cell lineage, defined according to the WHO classification system, for the presence of EBV. All tumors were negative for B-cell antigens, including PAX-5/BSAP and CD20 or CD79a. The study group included 27 (42%) anaplastic lymphoma kinase (ALK)-positive (18 T-cell and 9 null-cell) and 37 (58%) ALK-negative (30 T-cell and 7 null-cell) tumors analyzed by in situ hybridization for EBV-encoded RNA (EBER) or immunohistochemistry for EBV-latent membrane protein type 1. All 64 cases were negative for EBV. We conclude, based on the current definition of ALCL in the WHO classification, there is no role for EBV in ALCL arising in Western patients. We suggest that published reports of EBV in a small proportion of ALCL cases in Western patients can be explained by the inclusion of tumors no longer considered to be in the current classification of ALCL, such as CD30-positive anaplastic tumors of B-cell origin. PMID- 15116327 TI - Loss of heterozygosity on chromosome 9q22.3 in microdissected basal cell carcinomas around the Semipalatinsk Nuclear Testing Site, Kazakhstan. AB - A high incidence of skin cancers has been noted around the Semipalatinsk Nuclear Testing Site (SNTS) in Kazakhstan. Recently, basal cell carcinoma (BCC) susceptibility genes, human homolog of the Drosophila pathed gene (PTCH), and the xeroderma pigmentosa group A-complementing gene (XPA), have been cloned and localized on chromosome 9q22.3. To clarify the effect of low-dose irradiation on the occurrence of BCC, we used microdissection and polymerase chain reaction to identify loss of heterozygosity (LOH) at 9q22.3 using BCC samples obtained from this region. Ten Japanese samples were analyzed as controls. LOH with at least 1 marker was identified in 5 of 14 cases from around SNTS, whereas only 1 case with 1 marker was identified among the 10 Nagasaki cases. The total number of LOH alleles from SNTS (8 of 45) was significantly higher than the number from Nagasaki (1 of 26) (P = 0.03). The higher incidence of LOH on 9q22.3 in BCC from around SNTS suggests involvement of chronic low-dose irradiation by fallout from the test site as a factor in the cancers. PMID- 15116328 TI - Immunohistochemical detection of p53 homolog p63 in solid cell nests, papillary thyroid carcinoma, and hashimoto's thyroiditis: A stem cell hypothesis of papillary carcinoma oncogenesis. AB - Most models suggest that the cell of origin of papillary carcinoma is the mature thyroid follicular epithelial cell. In a recent study, p63 was detected in papillary carcinoma, Hashimoto's thyroiditis, and in squamoid aggregates and solid cell nests (SCNs), embryonic remnants found sporadically in the fully developed thyroid. In the present study, the relationship between solid cell nests and papillary carcinoma was investigated further. Four-micrometer sections from 88 routinely fixed and processed archival thyroidectomy specimens were pretreated with citric acid pH 6.0 for antigen retrieval, then incubated overnight with anti-p63 monoclonal antibody 4A4. Slides were stained with a streptavidin-biotin kit and diaminobenzidine as chromogen and were counterstained with hematoxylin. Squamoid aggregates or SCNs were noted in 21 specimens. Several morphologic variants of SCNs were found, all of which displayed p63 positivity. These included undifferentiated SCNs and those displaying commitment toward squamoid and ciliated glandular differentiation. Small, morphologically inconspicuous aggregates of p63-positive cells were commonly found in Hashimoto's thyroiditis. Commitment of p63-positive undifferentiated cells toward thyroid follicular epithelial differentiation was occasionally noted. One SCN variant, also associated with Hashimoto's thyroiditis, was a floretlike arrangement of p63 positive cells with fusiform nuclei. p63 staining was strong and uniform in some SCNs, but in other SCNs it was compartmentalized and homologous to stem cell staining patterns in normal squamous or bronchial epithelia. Stem cell-like staining, associated with compartmentalized p63 staining or p63-positive undifferentiated cells, was noted in 7 of 27 papillary carcinomas. p63 immunostaining is a highly sensitive means of detecting SCNs. p63 expression patterns in SCNs and a subset of papillary carcinomas are closely homologous to stem cell-associated p63 staining patterns that have been described elsewhere in squamous and bronchial epithelia. We propose a stem-cell-associated model of papillary carcinoma oncogenesis that suggests that (1) p63-positive embryonal remnants rather than mature follicular cells are the cells of origin of a subset of papillary carcinomas; (2) these p63-positive cells are pluripotent and may stay undifferentiated or undergo benign squamoid or glandular maturation, may undergo thyroid follicular epithelial differentiation, may undergo oncogenic change leading to papillary carcinoma, or may trigger an immune reaction, resulting in lymphoid infiltration and Hashimoto's thyroiditis; and (3) Hashimoto's thyroiditis and papillary carcinoma may therefore be linked etiologically, because both disorders may be initiated by the same population of pluripotent p63-positive embryonal stem cell remnants. PMID- 15116329 TI - Fungal sinusitis: histologic spectrum and correlation with culture. AB - Fungi are important etiologic agents of sinusitis. However, features of fungal sinusitis including the histologic spectrum, diagnostic mishaps, incidence, and fungal types have not been systematically studied. From 1996 through 2001, a total of 788 surgical pathology sinus specimens from 384 cases was retrieved. Fungal sinusitis was diagnosed in 58 specimens (7%) from 47 cases (12%). Four histologic categories of fungal sinusitis were identified: (1) allergic fungal sinusitis in 34 cases (copious mucin, abundant eosinophils, Charcot-Leyden crystals (so-called allergic mucin), with rare noninvasive fungal hyphae); (2) mycetoma/fungus ball in 11 cases (tightly packed fungal hyphae without allergic mucin or tissue invasion); (3) chronic invasive fungal sinusitis in 1 case (tissue granulomas with fungal hyphae); and (4) acute fulminant fungal sinusitis in 1 case (fungal vascular invasion). The diagnosis was initially missed in 16/34 (47%) cases of allergic fungal sinusitis despite typical features; incorrect classification was noted in 47% of cases. Sixty-seven percent of cases had positive fungal cultures, dematiaceous fungi being the most common. Allergic fungal sinusitis accounted for the majority of fungal sinusitis. Although misdiagnosis or incorrect classification is rather frequent for fungal sinusitis, awareness of the distinctive morphologic features of this entity may prevent these errors. PMID- 15116330 TI - Microdissection genotyping of archival fixative treated tissue for Gaucher disease. AB - The genetic diagnosis of Gaucher disease by molecular methods is complicated by the existence of a highly homologous transcribed pseudogene (96% identity) that is found in close proximity to the true gene on chromosome 1q21. In addition, the pseudogene sequence can mimic disease-causing mutations in the true gene. Selective polymerase chain reaction (PCR) amplification of the true gene can be accomplished in extracted DNA from fresh-frozen samples by designing oligonucleotide primers to hybridize to defined regions that are not present in the pseudogene. This standard molecular approach, which entails amplification of relatively long segments of intact DNA, is not feasible in archival, paraffin embedded, solid-tissue specimens in which the negative effects of chemical fixation result in DNA strand scission and breakdown of nucleic acid. A novel approach, specifically created for use with archival, fixative-treated tissue specimens, was developed for detection and characterization of common mutations of Gaucher disease. Three separate robust PCR reactions were formulated, 2 for selective amplification of portions of only the true gene exons 2 and 9, with a third reaction targeting exon 10, wherein both the true and pseudogene were coamplified. In the latter, DNA sequencing was used to determine the presence of true and pseudogene allele content in addition to identification of base sequence alterations. This method, requiring a single, 4-microm-thick histologic section, was successfully applied to archival paraffin block tissue specimens that had been in storage for up to 75 years. It was capable of accurately genotyping common Gaucher disease mutations as well as discovering a novel mutation and genetic polymorphism. We recommend our approach when only fixative-treated tis sue is available for molecular genotyping. PMID- 15116331 TI - Cyclooxygenase-2 expression correlates with angiogenesis and apoptosis in gastric cancer tissue. AB - In vitro studies suggest that cyclooxygenase-2 (COX-2) induces angiogenesis by stimulating angiogenic growth factors while inhibiting apoptosis in cancer cell lines. A series of 107 gastric adenocarcinoma cases that had undergone gastrectomy was studied to determine the correlation between COX-2 expression, angiogenesis, and apoptosis in human gastric cancer tissue. COX-2, vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and Bcl 2 were stained by single and dual immunoassaying methods. Microvessel density was determined by immunostaining for CD34. Apoptosis was evaluated with the TUNEL assay. COX-2 expression was positive exclusively in cancer cells in 46 cases (43%). COX-2 expression significantly correlated with VEGF and PDGF expression. Dual staining for COX-2 and VEGF showed that colocalization of these proteins was most frequent at the advancing edge of cancer cells. Microvessel density was higher in COX-2-and VEGF-positive cases than in COX-2- and VEGF-negative cases. In addition, COX-2 expression correlated with Bcl-2 expression. The apoptotic index was lower in COX-2-positive cancer cells than in COX-2-negative cases. Multivariate analysis revealed that coexpression of COX-2 and VEGF, age, lymph node status, and serosal invasion were independent prognostic factors for overall survival in gastric cancer patients. Therefore, these data suggest that COX-2 contributes to gastric cancer development by promoting angiogenesis and inhibiting apoptosis. PMID- 15116332 TI - Vasculogenic mimicry has no prognostic significance in pT3 and pT4 cutaneous melanoma. AB - The concept of vasculogenic mimicry has been introduced to define periodic acid Schiff (PAS)-positive channels and loops lined by tumor cells, instead of endothelium, able to contribute to microcirculation in uveal melanomas. Previous studies have shown that the PAS-positive patterns are associated with a poor prognosis in uveal melanoma. The aim of the current study was to investigate whether vasculogenic mimicry has a prognostic impact in pT3 and pT4 cutaneous melanoma. Fifteen patients with pT3 and pT4 cutaneous melanoma who did not experience progression after 10 years of follow-up and 30 matched controls who underwent progression were selected. Tumor sections were stained with PAS reaction, omitting the nuclear counterstaining. For immunohistochemistry, sections were stained with CD31, CD105 (endoglin), and laminin. Differences in the distribution of the PAS-positive patterns and a series of clinicopathological variables were evaluated by the Pearson chi(2) and Mann-Whitney U tests. We observed PAS-positive linear sheets, arcs, elliptical loops, and networks encircling roundish to oval aggregates of melanoma cells. The overall distribution of the PAS-positive patterns did not match with the blood microvessels' architecture as detected by immunohistochemical analysis. No statistically significant differences in the distribution of PAS-positive patterns were found between cases and controls. The presence of a parallel pattern correlated significantly with thickness (P = 0.04), whereas an inverse correlation was found with vessel area (P = 0.05). In conclusion, our results suggest that there is a mismatch between vasculogenic mimicry and tumor angiogenesis and do not support any prognostic role of vasculogenic mimicry in thick cutaneous melanoma. PMID- 15116333 TI - Immunohistochemical and ultrastructural markers suggest different origins for cuboidal and polygonal cells in pulmonary sclerosing hemangioma. AB - We report the morphological characteristics of 30 cases of sclerosing hemangioma (SH) of the lung and explore the histological origin of the major cells in these tumors. In addition to routine light and electron microscopy, immunohistochemistry was performed by using 12 monoclonal primary and 5 polyclonal primary antibodies. These included surfactant protein B (SP-B), thyroid transcription factor-1 (TTF-1), mast cell trypsin, CD68, epithelial antigen markers (high molecular weight cytokeratin, low molecular weight cytokeratin [CK-L], epithelial membrane antigen [EMA], cancer embryonic antigen), mesothelial antigen, neuroendocrine markers (neuron-specific enolase [NSE], chromogranin A, synaptophysin, calcitonin, adrenocorticotropic hormone, human growth hormone [hHG]), vimentin, and CD34. Surface cuboidal cells have short microvilli and have lamellar bodies in their cytoplasm. They can sometimes merge into multinuclear giant cells. Immunohistochemical results showed that these cells are strongly positive for SP-B, TTF-1, CK-L, EMA, and cancer embryonic antigen, whereas polygonal cells, previously also described as round or pale cells, were strongly positive for vimentin and TTF-1, and positive or weakly positive for 2 to 3 kinds of neuroendocrine markers. Sparse neuroendocrine granules and abundant microfilaments were observed in their cytoplasm. Some cell clusters in the solid regions were positive for SP-B and EMA. Mast cells existed sparsely in almost every field. Both cuboidal and polygonal cells were negative to CD34 and mesothelial antigen staining. We conclude that cuboidal cells of SH originate from reactive proliferating type II pneumocytes, which can fuse into multinuclear giant cells. Polygonal cells, as true tumor cells, likely originate from multipotential primitive respiratory epithelium and possess the capability for multipotential differentiation. The antibodies of SP-B, TTF-1, vimentin, and CK-L are very helpful to diagnosis and differential diagnosis of SH. PMID- 15116334 TI - Histological changes after the use of mycophenolate mofetil in autoimmune hepatitis. AB - Corticosteroids and azathioprine are considered standard treatment for autoimmune hepatitis. There are, however, well-known systemic side effects that may preclude continuation with these immunosuppressive therapies, and small trials of the anti metabolite mycophenolate mofetil have been reported. We report liver biopsy findings in a follow-up biopsy of a patient who had been switched from azathioprine to mycophenolate mofetil for treatment of presumed, steroid responsive autoimmune hepatitis. Both cytoplasmic features of adaptation and nuclear alterations were noted in hepatocytes. Possible mechanisms for these findings are discussed. PMID- 15116335 TI - Gliosarcoma arising from an anaplastic ependymoma: a case report of a rare entity. AB - We report the first case of a gliosarcoma arising from an anaplastic ependymoma and the second case of gliosarcoma arising from any type of ependymal neoplasm. The patient was a 48-year-old woman with a solid and cystic, peripherally enhancing, 7-cm right frontal mass lesion. Histologically, the lesion displayed characteristics of anaplastic ependymoma (grade III, World Health Organization scale). The tumor recurred despite multiple cycles of postoperative radiation and chemotherapy. After the fourth recurrence, the tumor displayed a biphasic pattern of differentiation. The first pattern was similar to the original anaplastic ependymoma, whereas there was a second new sarcomatous pattern resembling fibrosarcoma that was admixed with and overrunning the ependymal component. The spectrum of gliosarcoma is therefore expanded to include not only astrocytic and oligodendroglial components but ependymal components as well. PMID- 15116336 TI - Placental transmogrification of the lung: clinicopathologic, immunohistochemical and molecular study of two cases, with particular emphasis on the interstitial clear cells. AB - Two cases of placental transmogrification of the lung are reported. The lesions presented in the left lung, in one case as a giant bulla of the upper lobe and in the other as a cystic nodule of the lower lobe. A segmentectomy was performed in both cases, and the patients were alive and well 5 years and 2 months after surgery, respectively. In our opinion, pulmonary placental transmogrification is not a variant of emphysema, as generally considered, but rather probably represents a benign proliferation of immature interstitial clear cells with secondary cystic change. This report presents a histological, immunohistochemical, ultrastructural and molecular study of these peculiar cells, together with a review of the literature. PMID- 15116337 TI - Primary diagnosis of Whipple's disease in bone marrow. AB - Whipple's disease (WD) is a chronic systemic inflammatory disease of infectious origin caused by Tropheryma whipplei (TW). Abdominal pain and recurrent diarrhea are usually the main symptoms leading to the suspicion of a primary bowel disease. Systemic manifestations can mimic hematologic disorders. A 49-year-old man presented with fever, weight loss, long-standing arthralgia, and diarrhea. A duodenal biopsy was unremarkable. Bone marrow histology provided no evidence of a malignant hematological disorder but revealed noncaseating granulomas. TW was detected in the bone marrow trephine by polymerase chain reaction. This is the first report to describe TW-associated granulomatous myelitis as the initially recognized organ manifestation of WD, proven at the molecular level. This observation is relevant for the differential diagnosis of patients with systemic symptoms and granulomatous diseases affecting the bone marrow, emphasizing that WD should be considered in cases of unexplained granulomatous myelitis, even when small bowel biopsy specimens are negative. PMID- 15116338 TI - Intraprostatic fat: does it exist? PMID- 15116339 TI - Structural analysis of glucoamylase encoded by the STA1 gene of Saccharomyces cerevisiae (var. diastaticus). AB - The sequence of the STA1-encoded glucoamylase of amylolytic Saccharomyces cerevisiae (var. diastaticus) strains shows two well-defined regions: an amino terminal part rich in serine and threonine residues and a carboxy-terminal part very similar to the catalytic domain of other fungal glucoamylases. A version of the enzyme in which most of the amino-terminal region was deleted still has glucoamylase activity, indicating that the remaining carboxy-terminal part forms a functional catalytic domain. Homology-based models of the two parts of the protein have been obtained. As expected, the shortened form of the enzyme is very similar to the catalytic domain of related glucoamylases of known structure. However, the amino-terminal part yielded a structure revealing an unexpected similarity to bacterial invasins, suggesting functional connections between several yeast proteins homologous to STA1-encoded glucoamylase and invasins. A characteristic of Saccharomyces glucoamylase in its native form is its extreme degree of glycosylation. Despite its high molecular mass (about 300 kDa), and in contrast with what occurs with other extracellular glycoproteins produced by yeast, the enzyme does not remain attached to the cell wall, being fully and efficiently secreted into the medium, even when it is produced in large amounts by overexpression of its gene. PMID- 15116340 TI - The absence of the Isw2p-Itc1p chromatin-remodelling complex induces mating type specific and Flo11p-independent invasive growth of Saccharomyces cerevisiae. AB - The Isw2p-Itc1p chromatin remodelling complex of Saccharomyces cerevisiae is a member of the ISWI class of ATPases with a nucleosome spacing activity, involved in regulation of expression of a broad spectrum of genes. Its absence causes derepression of a-specific genes and aberrant morphology in alpha-mating type cells. We report here that the deletion of the ISW2 gene in the originally non invasive BY strain induces mating type-specific invasive growth strongly affected by nitrogen starvation. Although the Flo11 protein was postulated to be critical for haploid invasive growth, we showed that the invasive growth caused by the isw2 and itc1 deletions in alpha-mating type cells was Flo11p-independent. This type of invasive growth was proved to be a consequence of the activation of the pheromone response pathway. Our results suggest that Isw2 and Itc1 proteins do not have the same impact on the described phenomenon. PMID- 15116341 TI - Sodium and potassium transport in the halophilic yeast Debaryomyces hansenii. AB - Debaryomyces hansenii, a halophile yeast found in shallow sea waters and salty food products grows optimally in 0.6 M of either NaCl or KCl, accumulating high concentrations of Na(+) or K(+). After growth in NaCl or KCl, a rapid efflux of either accumulated cation was observed if the cells were incubated in the presence of KCl or NaCl, respectively, accompanied by a slower accumulation of the cation present in the incubation medium. However, a similar, rapid efflux was observed if cells were incubated in buffer, in the absence of external cations. This yeast shows a cation uptake activity of both (86)Rb(+) and (22)Na(+) with saturation kinetics, and much higher affinity for (86)Rb(+) than for (22)Na(+). The pH dependence of the kinetics constants was similar for both cations, and although K(m) values were higher at pH 8.0, there was also an increase in the V(max) values. The accumulation of (22)Na(+) was found to be increased in cells grown in the presence of 0.6 M NaCl. (86)Rb(+) was also accumulated more in these cells, but to a slightly greater extent. The inhibition kinetics of the uptake of (22)Na(+) by K(+), and that of (86)Rb(+) by Na(+) was found to be non competitive. It can be concluded that Na(+) in D. hansenii is not excluded but instead, its metabolic systems must be resistant to high salt concentrations. PMID- 15116342 TI - Characterization of the transcriptional response to cell wall stress in Saccharomyces cerevisiae. AB - The cell wall perturbants Calcofluor white and Zymolyase activate the Pkc1-Rho1 controlled Slt2p MAP kinase pathway in Saccharomyces cerevisiae. A downstream transcription factor of this pathway, Rlm1p, is known to control expression of about 20 cell wall-related genes. Global transcript analysis of Calcofluor white and Zymolyase treatment was performed to determine whether cell wall stress affects transcription of these and other genes. Transcript profiles were analysed using two recently developed algorithms, viz. REDUCE, which correlates upstream regulatory motifs with expression, and Quontology, which compares expression of genes from functional groups with overall gene expression. Both methods indicated upregulation of Rlm1p-controlled cell wall genes and STRE-controlled genes, and downregulation of ribosomal genes and rRNA genes. Comparison of these expression profiles with the published profiles of two constitutively active upstream activators of the Slt2p-MAP kinase pathway, viz. Pkc1-R398A and Rho1-Q68A, revealed significant similarity. In addition, a new putative regulatory motif, CCC(N)(10)GGC, was found. In Zymolyase-treated cells a regulatory site was identified, ATGACGT, which resembles the AFT/CRE binding site. Interestingly, Sko1p, a downstream regulator of the high osmolarity pathway is known to bind to the AFT/CRE binding site, suggesting a possible role for the Hog1 pathway in the response to cell wall stress. Finally, using REDUCE, an improved version of the Rlm1 binding motif, viz. TA(W)(4)TAGM, was discovered. We propose that this version can be used in combination with REDUCE as a sensitive indicator of cell wall stress. Taken together, our data indicate that cell wall stress results in activation of various signalling pathways including the cell wall integrity pathway. PMID- 15116343 TI - Cassettes for the PCR-mediated construction of regulatable alleles in Candida albicans. AB - The recent availability of genome sequence information for the opportunistic pathogen Candida albicans has greatly facilitated the ability to perform genetic manipulations in this organism. Two important molecular tools for studying gene function are regulatable promoters for generating conditional mutants and fluorescent proteins for determining the subcellular localization of fusion gene products. We describe a set of plasmids containing promoter-GFP cassettes (P(MET3)-GFP, P(GAL1)-GFP, and P(PCK1)-GFP), linked to a selectable nutritional marker gene (URA3). PCR-mediated gene modification generates gene-specific promoter, or gene-specific promoter-GFP, fusions at the 5'-end of the gene of interest. One set of primers can be used to generate three strains expressing a native protein of interest, or an amino-terminal GFP-tagged version, from three different regulatable promoters. Thus, these promoter cassette plasmids facilitate construction of conditional mutant strains, overexpression alleles and/or inducible amino-terminal GFP fusion proteins. PMID- 15116344 TI - Cloning and functional characterization of the SUR2/SYR2 gene encoding sphinganine hydroxylase in Pichia ciferrii. AB - Saccharomyces cerevisiae sphinganine C4-hydroxylase encoded by the SUR2 gene catalyses the conversion of sphinganine to phytosphingosine. We isolated the SUR2 gene from Pichia ciferrii using nucleotide sequence homology to S. cerevisiae SUR2 to study hydroxylation of sphinganine in the sphingoid base overproducing yeast P. ciferrii. A positive clone was confirmed by nucleotide sequencing. A syringomycin-E resistance phenotype of a S. cerevisiae sur2-null mutant was complemented by expression of the cloned P. ciferrii SUR2 gene. Restoration of phytosphingosine production in the complemented strain was also confirmed, indicating that the cloned gene is a functional homologue of S. cerevisiae SUR2. . PMID- 15116345 TI - Molecular characterization of the glutathione-dependent formaldehyde dehydrogenase gene FLD1 from the methylotrophic yeast Pichia methanolica. AB - In this paper, we describe molecular characterization of the FLD1 gene, which encodes glutathione-dependent formaldehyde dehydrogenase (FLD), from the methylotrophic yeast Pichia methanolica. The P. methanolica FLD1 gene contains two exons corresponding to a gene product of 380 amino acid residues and a 225 bp intron, respectively, and its deduced amino acid sequence shows high similarity to those of Fld1ps from other methylotrophic yeasts (80-88%). In P. methanolica, FLD activity is mainly induced by methanol, and this induction is not completely repressed by glucose. Moreover, the expression of the PmFLD1 is strictly regulated, mainly at the mRNA level, its expression increasing with increasing methanol concentrations in the medium. These results suggest that FLD1 is involved in the detoxification of formaldehyde in methanol metabolism, and Fld1p coordinates the formaldehyde level in methanol-grown cells according to the methanol concentration on growth. PMID- 15116346 TI - Using Latent Mixed Markov Models for the choice of the best pharmacological treatment. AB - The choice of the best pharmacological treatment for an individual patient is crucial to optimize convalescence. Due to their effects on pharmacokinetics variables like gender and age are important factors when the pharmacological regimen is planned. By means of an example from anaesthesiology the usefulness of Latent Mixed Markov Models for choosing the optimal anaesthetic considering patient characteristics is demonstrated. Latent Mixed Markov models allow to predict and compare the quality of recovery from anaesthesia for different patient groups (defined by age and gender and treated with different anaesthetic regimens) in a multivariate non-parametric approach. On the basis of observed symptoms immediately after surgery and a few days later the probabilities for the respective dynamic latent status (like health or illness) and the probabilities for transition from one status to another are estimated depending on latent class membership (patient group). PMID- 15116347 TI - What to add to nothing? Use and avoidance of continuity corrections in meta analysis of sparse data. AB - OBJECTIVES: To compare the performance of different meta-analysis methods for pooling odds ratios when applied to sparse event data with emphasis on the use of continuity corrections. BACKGROUND: Meta-analysis of side effects from RCTs or risk factors for rare diseases in epidemiological studies frequently requires the synthesis of data with sparse event rates. Combining such data can be problematic when zero events exist in one or both arms of a study as continuity corrections are often needed, but, these can influence results and conclusions. METHODS: A simulation study was undertaken comparing several meta-analysis methods for combining odds ratios (using various classical and Bayesian methods of estimation) on sparse event data. Where required, the routine use of a constant and two alternative continuity corrections; one based on a function of the reciprocal of the opposite group arm size; and the other an empirical estimate of the pooled effect size from the remaining studies in the meta-analysis, were also compared. A number of meta-analysis scenarios were simulated and replicated 1000 times, varying the ratio of the study arm sizes. RESULTS: Mantel-Haenszel summary estimates using the alternative continuity correction factors gave the least biased results for all group size imbalances. Logistic regression was virtually unbiased for all scenarios and gave good coverage properties. The Peto method provided unbiased results for balanced treatment groups but bias increased with the ratio of the study arm sizes. The Bayesian fixed effect model provided good coverage for all group size imbalances. The two alternative continuity corrections outperformed the constant correction factor in nearly all situations. The inverse variance method performed consistently badly, irrespective of the continuity correction used. CONCLUSIONS: Many routinely used summary methods provide widely ranging estimates when applied to sparse data with high imbalance between the size of the studies' arms. A sensitivity analysis using several methods and continuity correction factors is advocated for routine practice. PMID- 15116348 TI - GEE approaches to marginal regression models for medical diagnostic tests. AB - The evaluation of a new medical diagnostic test may focus on two different scientific questions: (1). The new test may replace an existing one because of lower cost or higher validity. A related question would be the selection of the 'best' test(s) from a bundle of new or established measurements. (2). The new test may be used supplementary to other new or established procedures. In a recent publication, Leisenring and co-workers (Stat Med 1997; 16:1263-1281) developed a general marginal regression model for comparisons of diagnostic tests focussing on question (1). i.e. on the selection of the 'best' procedure. They applied the GEE approach of Liang and Zeger (Biometrika 1987; 73:13-22) to adjust for the correlation of data as a nuisance parameter. Using the general framework provided by Leisenring et al., we extend their approach and apply the GEE methodology to question (2). i.e. to the investigation of which of several diagnostic tests should be used supplementary to each other. We analyse data from a longitudinal study concerning pathogenesis, diagnosis and long-term course of the eye disease glaucoma. We find a dependence of the correlation structure of several diagnostic measurements on the severity of the disease. This result may be useful in clinical applications as regards the selection of subsets of diagnostic measurements in individual diagnostic processes but also in investigations concerning the relationship of the pathogenic process and the rationales of the different diagnostic procedures. PMID- 15116349 TI - The 'Fishing License' method for analysing the time course of effects in repeated measurements. AB - Inference on the time of onset and the duration of a treatment effect is a challenging problem in biomedical research. These studies often generate repeated measurements from subjects in a treated group and a control group during the same time period. We propose a simple approach, called the 'Fishing License' method, to test for a treatment effect occurring during an unspecified time interval. The method is based on a statistic of the largest absolute value of the t statistic between two groups obtained by considering every possible time interval in the observed time period. A bootstrapped null distribution of the test statistic is used to determine the critical value. The method also provides estimates of onset and ending times, when the null hypothesis of no effect is rejected. Simulated and real experimental data sets were generated for assessing the performance of the method. PMID- 15116350 TI - Construction of group sequential designs in clinical trials on the basis of detectable treatment differences. AB - The treatment effect sizes that can be detected with sufficient power up to the different interim analyses constitute a clinically meaningful criterion for the selection of a group sequential test for a clinical trial. For any pre-specified sequence of effect sizes, it is possible to construct group sequential boundaries such that the trial has a pre-specified power to reject the null-hypothesis at or before the corresponding interim analysis under the respective treatment effect. The principle of constructing group sequential designs on the basis of detectable treatment effects is presented. The application in common situations such as two armed trials with continuous or binary outcome or censored survival times is described. We also present an effective algorithm. PMID- 15116351 TI - A tree based lack-of-fit test for multiple logistic regression. AB - Several omnibus tests have been developed to assess the fit of a regression model. But many of these lack-of-fit tests focus on the simple regression setting. Here, we focus on multiple logistic regression. Pearson's well-known chi square test statistic and the deviance statistic are no longer valid in the case that the model contains one or more continuous covariates. To overcome this difficulty, Hosmer and Lemeshow proposed a Pearson type statistic based on groups defined by the so-called deciles of risk. We propose a test statistic that is similar in approach to the Hosmer and Lemeshow statistic in that the observations are classified into distinct groups. In the procedure proposed here however, the grouping is not according to probabilities fitted under the null model. We use a recursive partitioning algorithm to divide the sample space into different groups. This generally allows for a more powerful assessment of the model fit. Simulations are carried out to compare the results of the proposed test to that of Hosmer and Lemeshow. Three data examples illustrate the performance of the tree based lack-of-fit test, in comparison to several other tests. PMID- 15116352 TI - Tree and spline based association analysis of gene-gene interaction models for ischemic stroke. AB - In the biology of complex disorders, such as atherothrombosis, interactions among genetic factors may play an important role, and theoretical considerations suggest that gene-gene interactions are quite common in such diseases. We used a nested case-control sample from the Physicians' Health Study, a randomized trial assessing the effects of aspirin and beta-carotene on cardiovascular disease and cancer among 22071 US male physicians, to examine these relationships for ischemic stroke. Data were available on 92 polymorphisms from 56 candidate genes related to inflammation, thrombosis and lipid metabolism, assessed in 319 incident cases of ischemic stroke and 2090 disease-free controls. We used classification and regression trees (CART) and multivariate adaptive regression spline (MARS) models to explore the presence of genetic interactions in these data. These models offer advantages over typical logistic regression methods in that they may uncover interactions among genes that do not exhibit strong marginal effects. Final models were selected using either the Bayes Information Criterion or cross-validation. Model fit was assessed using 10-fold cross validation of the entire selection process. Both the CART and two-way MARS-logit models identified an interaction between two polymorphisms linked to inflammation, the P-selectin (val640leu) and interleukin-4 (C(582) T) genes. Internal validation of these models, however, suggested that effects of these polymorphisms are additive. Although further external validation of these models is necessary, these methods may be valuable in exploring and identifying potential gene-gene as well as gene-environment interactions in association studies. PMID- 15116353 TI - Handling drop-out in longitudinal studies. AB - Drop-out is a prevalent complication in the analysis of data from longitudinal studies, and remains an active area of research for statisticians and other quantitative methodologists. This tutorial is designed to synthesize and illustrate the broad array of techniques that are used to address outcome-related drop-out, with emphasis on regression-based methods. We begin with a review of important assumptions underlying likelihood-based and semi-parametric models, followed by an overview of models and methods used to draw inferences from incomplete longitudinal data. The majority of the tutorial is devoted to detailed analysis of two studies with substantial rates of drop-out, designed to illustrate the use of effective methods that are relatively easy to apply: in the first example, we use both semi-parametric and fully parametric models to analyse repeated binary responses from a clinical trial of smoking cessation interventions; in the second, pattern mixture models are used to analyse longitudinal CD4 counts from an observational cohort study of HIV-infected women. In each example, we describe exploratory analyses, model formulation, estimation methodology and interpretation of results. Analyses of incomplete data requires making unverifiable assumptions, and these are discussed in detail within the context of each application. Relevant SAS code is provided. PMID- 15116354 TI - Geometry optimization in density functional methods. AB - The geometry optimization in delocalized internal coordinates is discussed within the framework of the density functional theory program deMon. A new algorithm for the selection of primitive coordinates according to their contribution to the nonredundant coordinate space is presented. With this new selection algorithm the excessive increase in computational time and the deterioration of the performance of the geometry optimization for floppy molecules and systems with high average coordination numbers is avoided. A new step selection based on the Cartesian geometry change is introduced. It combines the trust radius and line search method. The structure of the new geometry optimizer is described. The influence of the SCF convergence criteria and the grid accuracy on the geometry optimization are discussed. A performance analysis of the new geometry optimizer using different start Hessian matrices, basis sets and grid accuracies is given. PMID- 15116355 TI - Evaluation of the protein solvent-accessible surface using reduced representations in terms of critical points of the electron density. AB - The aim of our study is the development of a method for calculating the interface of dimerization of protein-protein complexes based on simplified medium resolution structures. In particular, we wished to evaluate if the existing concepts for the computation of the Solvent-Accessible Surface Area (SASA) of macromolecules could be applied to medium-resolution models. Therefore, we selected a set of 140 protein chains and computed their reduced representations by topological analysis of their electron density maps at 2.85 A crystallographic resolution. This procedure leads to a limited number of critical points (CPs) that can be identified and associated to backbone and side-chain parts. To evaluate the SASA and interfaces of dimerization of the reduced representations, we chose and modified two existing programs that calculate the SASA of atomic representations, and tested (1) several radii tables of amino acids, (2) the influence of the backbone and side-chain points, and (3) the radius of the solvent molecule, which rolls over the surface. The results are shown in terms of relative error compared to the values calculated on the corresponding atomic representations of the proteins. PMID- 15116356 TI - Conformational energies for 2-substituted butanes. AB - The conformational free energies for some 2-substituted butanes where X = F, Cl, CN, and CCH were calculated using G3-B3, CBS-QB3, and CCSD(T)/6-311++G(2d,p) as well as other theoretical levels. The above methods gave consistent results with free energies relative to the trans conformers as follows: X = CCH, g+ = 0.77 +/- 0.05 kcal/mol. g- = 0.88 +/- 0.05 kcal/mol; X = CN, g+ = 0.85 +/- 0.05 kcal/mol, g- = 0.75 +/- 0.05 kcal/mol; X = Cl, g+ = 0.70 +/- 0.05 kcal/ml, g- = 0.80 +/- 0.05 kcal/mol; and X = F, g+ = 0.53 +/- 0.05 kcal/mol, g- = 0.83 +/- 0.05 kcal/mol. The conformational free energies also were estimated using the observed liquid phase IR spectra and intensities calculated using B3LYP/6-311++G** and MP2/6-311++G**. The rotational free energy profiles for all of the compounds were estimated at the G3-B3 level. PMID- 15116357 TI - Sampling protein conformations and pathways. AB - Protein flexibility and rigidity can be analyzed using constraint theory, which views proteins as 3D networks of constraints involving covalent bonds and also including hydrophobic interactions and hydrogen bonds. This article describes an algorithm, ROCK (Rigidity Optimized Conformational Kinetics), which generates new conformations for these complex networks with many interlocked rings while maintaining the constraints. These new conformations are tracked for the flexible regions of a protein, while leaving the rigid regions undisturbed. An application to HIV protease demonstrates how large the flap motion can be. The algorithm is also used to generate conformational pathways between two distinct protein conformations. As an example, directed trajectories between the closed and the occluded conformations of the protein dihydrofolate reductase are determined. PMID- 15116358 TI - Multicut-HDMR with an application to an ionospheric model. AB - A new High Dimensional Model Representation (HDMR) tool, Multicut-HDMR, is introduced and applied to an ionospheric electron density model. HDMR is a general set of quantitative model assessment and analysis tools for improving the efficiency of deducing high-dimensional input-output system behavior. HDMR describes an output [f(x)] in terms of its input variables (x = [x(1), x(2), em leader, x(n)]) via a series of finite, hierarchical, correlated function expansions. Various forms of HDMR are constructed for different purposes such as modeling laboratory or field data, or reproducing a complicated mathematical model. The Cut-HDMR technique, which expresses f(x) with respect to a specified reference point x in the input space, is appropriate when the input space is sampled in an orderly fashion. However, if the desired domain of the input space is too large, the HDMR function expansion may not converge, and Cut-HDMR will be unable to accurately approximate f(x). The new Multicut-HDMR technique addresses this problem through the use of multiple reference points in the input space. PMID- 15116359 TI - Development and testing of a general amber force field. AB - We describe here a general Amber force field (GAFF) for organic molecules. GAFF is designed to be compatible with existing Amber force fields for proteins and nucleic acids, and has parameters for most organic and pharmaceutical molecules that are composed of H, C, N, O, S, P, and halogens. It uses a simple functional form and a limited number of atom types, but incorporates both empirical and heuristic models to estimate force constants and partial atomic charges. The performance of GAFF in test cases is encouraging. In test I, 74 crystallographic structures were compared to GAFF minimized structures, with a root-mean-square displacement of 0.26 A, which is comparable to that of the Tripos 5.2 force field (0.25 A) and better than those of MMFF 94 and CHARMm (0.47 and 0.44 A, respectively). In test II, gas phase minimizations were performed on 22 nucleic acid base pairs, and the minimized structures and intermolecular energies were compared to MP2/6-31G* results. The RMS of displacements and relative energies were 0.25 A and 1.2 kcal/mol, respectively. These data are comparable to results from Parm99/RESP (0.16 A and 1.18 kcal/mol, respectively), which were parameterized to these base pairs. Test III looked at the relative energies of 71 conformational pairs that were used in development of the Parm99 force field. The RMS error in relative energies (compared to experiment) is about 0.5 kcal/mol. GAFF can be applied to wide range of molecules in an automatic fashion, making it suitable for rational drug design and database searching. PMID- 15116360 TI - Improvement of parallelization performance of GAMESS: global sum and (semi )direct integral calculation in multireference perturbation calculation. AB - Performance benchmark tests of the multireference perturbation theory (MRPT2) calculation in the GAMESS suite of programs have been performed on a 128-node PC cluster system. The benchmark results point to two kinds of problems with GAMESS: network delay and slow access to disk files. To overcome these bottlenecks, we employ (semi-)direct atomic orbital integral calculations and a new algorithm for the "global sum" operation. These modifications produce remarkable performance improvements, especially in the 128-node PC cluster system, which was almost twice as fast in our version as in the original version. The modification of "global sum" also improved the performance of other types of molecular orbital calculations in GAMESS, such as density functional theory. PMID- 15116361 TI - Theoretical study on the mechanism of the 1CHCl + NO2 reactions. AB - The radical-molecule reaction mechanism of (1)CHCl with NO(2) has been explored theoretically at the B3LYP/6-311G(d, p) and CCSD(T)/6-311G(d, p) (single-point) levels of theory. Thirteen minimum isomers and 29 transition states are located. The initial association between (1)CHCl and NO(2) proceeds most likely through the carbon-to-middle-nitrogen attack leading to an energy-rich adduct a (HClCNO(2)), which is found to be a barrierless process. Staring from a, the most feasible channel is to undergo a concerted O-shift and C--N bond rupture leading to product P(2) (NO + HClCO). The minor product pathways are the direct O extrusion of a to P(3) (O + HClCNO-cis) as well as the 1,3-H-shift of a to isomer b (ClCNOOH) followed by a concerted OH-shift leading to d (HOClCNO), which will dissociate to product P(8) (NO + ClCOH) via C--N cleavage. Because the transition states and isomers involved in the most feasible channel all lie below the reactants, the title reaction is expected to be rapid, as is consistent with the measured rate constant at 296 K. The comparison with the analogous reactions (3)CH(2) + NO(2) are discussed. The present study may be useful for further experimental investigation of the title reaction. PMID- 15116362 TI - A precise boundary element method for macromolecular transport properties. AB - A very precise boundary element numerical solution of the exact formulation of the hydrodynamic resistance problem with stick boundary conditions is presented. BEST, the Fortran 77 program developed for this purpose, computes the full transport tensors in the center of resistance or the center of diffusion for an arbitrarily shaped rigid body, including rotation-translation coupling. The input for this program is a triangulation of the solvent-defined surface of the molecule of interest, given by Connolly's MSROLL or other suitable triangulator. The triangulation is prepared for BEST by COALESCE, a program that allows user control over the quality and number of triangles to describe the surface. High numerical precision is assured by effectively exact integration of the Oseen tensor over triangular surface elements, and by scaling the hydrodynamic computation to the precise surface area of the molecule. Efficiency of computation is achieved by the use of public domain LAPACK routines that call BLAS Level 3 hardware-optimized subroutines available for most processors. A protein computation can be done in less than 10 min of CPU time in a modern Pentium IV processor. The present work includes a complete analysis of the sources of error in the numerical work and techniques to eliminate these errors. The operation of BEST is illustrated with applications to ellipsoids of revolution, and Lysozyme, a small protein. The typical numerical accuracy achieved is 0.05% compared to analytical theory. The numerical precision for a protein is better than 1%, much better than experimental errors in these quantities, and more than 10 times better than traditional bead-based methods. PMID- 15116363 TI - Improved model core potentials for the second- and third-row transition metals. AB - New nonrelativistic and scalar-relativistic pseudopotentials for the second- and third-row transition metals have been developed. These improved Model Core Potentials were used in calculations for a variety of transition metal complexes to test their ability to reproduce experimental structures and vibrational frequencies. PMID- 15116364 TI - Sir John A. Pople, 1925-2004. PMID- 15116366 TI - Botulinum toxins in neurological disease. AB - Botulinum toxins are among the most potent neurotoxins known to humans. In the past 25 years, botulinum toxin has emerged as both a potential weapon of bioterrorism and as a powerful therapeutic agent, with growing applications in neurological and non-neurological disease. Botulinum toxin is unique in its ability to target peripheral cholinergic neurons, preventing the release of acetylcholine through the enzymatic cleavage of proteins involved in membrane fusion, without prominent central nervous system effects. There are seven serotypes of the toxin, each with a specific activity at the molecular level. Currently, serotypes A (in two preparations) and B are available for clinical use, and have been shown to be safe and effective for the treatment of dystonia, spasticity, and other disorders in which muscle overactivity gives rise to symptoms. This review focuses on the pharmacology, electrophysiology, immunology, and application of botulinum toxin in selected neurological disorders. PMID- 15116365 TI - Structural and functional changes in spastic skeletal muscle. AB - This review summarizes current information regarding the changes in structure or function that occur in skeletal muscle secondary to spasticity. Most published studies have reported an increase in fiber size variability in spastic muscle. There is no general agreement regarding any shift in fiber type distribution secondary to spasticity. Mechanical studies in whole limbs as well as in isolated single cells support the notion of an intrinsic change in the passive mechanical properties of muscle after spasticity in addition to the more widely reported neural changes that occur. Evidence is presented for changes within both the muscle cell and extracellular matrix that contribute to the overall changes in the tissue. Taken together, the literature supports the notion that, although spasticity is multifactorial and neural in origin, significant structural alterations in muscle also occur. An understanding of the specific changes that occur in the muscle and extracellular matrix may facilitate the development of new conservative or surgical therapies for this problem. PMID- 15116367 TI - Nerve excitability properties in lower-limb motor axons: evidence for a length dependent gradient. AB - In this study, nerve excitability protocols were adapted for lower-limb recordings in 25 healthy subjects to enable comparison of excitability parameters between proximal and distal recording sites of the same nerve and between different nerves. Excitability parameters (stimulus-response curves, strength duration properties, threshold electrotonus, a current-threshold relationship, and the recovery cycle) were recorded from tibialis anterior, extensor digitorum brevis, and abductor hallucis. Excitability recordings were technically possible from each site, and normative values were established for lower-limb nerves. In this process, inter- and intranerve differences in excitability properties were demonstrated: stimulus intensity and rheobase were reduced in recordings from proximal sites; the relative refractory period and late subexcitability were increased; superexcitability was reduced; and a relative "fanning-in" occurred for threshold electrotonus curves recorded from proximal sites. Such a length dependent gradient in nerve excitability may underlie the greater tendency for ectopic activity to arise from the proximal segments of motor axons and may contribute to the length-dependent involvement of motor axons in the development of peripheral neuropathy. PMID- 15116368 TI - Effects of high-intensity endurance exercise training in the G93A mouse model of amyotrophic lateral sclerosis. AB - The G93A transgenic mouse has a mutation in copper/zinc superoxide dismutase (CuZnSOD) that results in oxidative stress and motor neuron loss. Endurance exercise training is known to increase antioxidant capacity in skeletal muscle. Therefore, we hypothesized that endurance training may extend onset of disease or survival in the G93A mouse. We examined the effects of high-intensity endurance exercise training (45 min/day, 5 times/week, progressive increase from 9 to 22 m/min) on disease onset and survival in G93A mice. Endurance training did not affect clinical onset, although it hastened death in male mice (P < 0.05). Endurance-trained males had a statistically significant decrease in rotarod performance at 112 days (P < 0.05), whereas sedentary males decreased at 119 days (P < 0.05). Endurance-trained and sedentary females decreased at 126 days and 129 days, respectively (P < 0.05). Female mice lived longer than males (P < 0.05), and there was a trend for hastened clinical onset in males (P = 0.062). We conclude that high-intensity endurance exercise training does not affect onset of clinical symptoms in G93A mice but hastens a decrease in motor performance and death following onset of clinical symptoms in male mice only. In light of a recent report describing increased survival following low-intensity endurance training, it appears that training intensity is an important determinant of survival in the G93A mouse. PMID- 15116369 TI - Different short-term modulation of cortical motor output to distal and proximal upper-limb muscles during painful sensory nerve stimulation. AB - The pattern of upper-limb muscle activation following painful stimulation has not been clarified in detail. We investigated the short-term inhibitory and excitatory effects of painful electrical digital stimulation on the motoneuron pools of distal and proximal upper-limb muscles. Transcranial magnetic stimulation (TMS) was used as test stimulus, and painful digital nerve stimulation as conditioning stimulus for motor evoked potential (MEP) recordings over the abductor digiti minimi (ADM), abductor pollicis brevis (APB), biceps brachii (BB), and deltoid muscles. Inhibition of the conditioned MEP response was most prominent in the distal muscles, whereas BB and deltoid muscles were only weakly inhibited. The mean MEP response over APB decreased with painful cutaneous stimuli, showing maximum inhibition (by 82%) at interstimulus intervals (ISIs) of 50 ms. Inhibition in the ADM was maximal (49%) but less pronounced at an ISI of 40 ms. The BB and deltoid muscles showed inhibition by 25% and 29%, respectively. Significant facilitation was present in BB and deltoid muscles by 43% and 41% at an ISI of 100 ms, but not in the smaller hand muscles. The observed pattern of upper-limb muscle activation corresponds to the protective withdrawal reflex and the neuronal basis of the observed short-term modulation of motor activity is compatible with a spinal or brainstem pathway. PMID- 15116370 TI - Exon 17 skipping in CLCN1 leads to recessive myotonia congenita. AB - Mutations in CLCN1, the gene encoding the ClC-1 chloride channel in skeletal muscle, lead to myotonia congenita. The effects on the intramembranous channel forming domains have been investigated more than that at the intracellular C terminus. We have performed a mutation screen involving the whole CLCN1 gene of patients with myotonia congenita by polymerase chain reaction (PCR), single strand conformation polymorphism studies, and sequencing. Two unrelated patients harbored the same homozygous G-to-T mutation on the donor splice site of intron 17. This led to the skipping of exon 17, as evidenced by the reverse transcriptase PCR. When the exon 17-deleted CLCN1 was expressed in Xenopus oocytes, no chloride current was measurable. This function could be restored by coexpression with the wild-type channel. Our data suggest an important role of this C-terminal region and that exon 17 skipping resulting from a homozygous point mutation in CLCN1 can lead to recessive myotonia congenita. PMID- 15116371 TI - Absence of sensory neuropathy among workers with occupational exposure to chlorpyrifos. AB - Several studies have reported the occurrence of sensory neuropathy with exposure to chlorpyrifos and other organophosphorus insecticides, at levels not associated with overt toxicity. We evaluated 113 chemical workers, including 53 of 66 (80%) eligible chlorpyrifos workers and 60 of 74 (81%) randomly selected referent workers, to identify evidence of sensory neuropathy or subclinical neuropathy. Compared to referents, chlorpyrifos subjects had significantly longer duration of work in chlorpyrifos-exposed areas (9.72 vs. 0.01 years; P < 0.0001), greater cumulative chlorpyrifos exposure (64.16 vs. 0.69 mg/m(3). day; P < 0.0001), higher urine 3,5,6-trichloro-2-pyridinol (TCP) excretion (108.6 vs. 4.3 microg/g creatinine; P < 0.0001), and lower plasma butyrylcholinesterase (BuChE) activity (7281 vs. 8176 mU/ml; P = 0.003). Despite exposures among chlorpyrifos subjects to levels at which well-described physiological effects on B-esterases exist, the frequency of symptoms or signs of neuropathy did not differ significantly between groups, and the only 2 subjects fulfilling criteria for confirmed neuropathy were both in the referent group. Mean nerve conduction study results were comparable to established control values and did not differ significantly between groups. We found no evidence of sensory neuropathy or isolated peripheral abnormalities among subjects with long-term chlorpyrifos exposure at levels known to be associated with the manufacturing process. PMID- 15116372 TI - Effect of creatine supplementation on skeletal muscle of mdx mice. AB - Dystrophic mice (mdx) and their controls (C57/Bl10) were fed for 1 month with a diet with or without creatine (Cr) enrichment. Cr supplementation reduced mass (by 19%, P < 0.01) and mean fiber surface (by 25%, P < 0.05) of fast-twitch mdx muscles. In both strains, tetanic tension increased slightly (9.2%) without reaching statistical significance (P = 0.08), and relaxation time increased by 16% (P < 0.001). However, Cr had no protective effect on the other hallmarks of dystrophy such as susceptibility to eccentric contractions; large numbers of centrally nucleated fibers in tibialis anterior; and elevated total calcium content, which increased by 85% (P = 0.008) in gastrocnemius mdx muscles. In conclusion, Cr may be a positive intervention for improving function of dystrophic muscle. PMID- 15116373 TI - Motor unit number estimation by decomposition-enhanced spike-triggered averaging: control data, test-retest reliability, and contractile level effects. AB - Decomposition-enhanced spike-triggered averaging (DE-STA) has been developed as a method for obtaining a motor unit number estimate (MUNE). We describe the method and report control data for the first dorsal interosseous/adductor pollicis and thenar muscles and reliability in the thenar muscles. Seventeen subjects (ages 20 50 years) took part in the study. The maximum M potential was elicited with supramaximal stimulation of the ulnar or median nerve at the wrist. Surface and intramuscularly detected electromyographic signals were then collected simultaneously during mild to moderate contractions. Decomposition algorithms were used to detect and sort the individual motor unit potential (MUP) occurrences of several concurrently active motor units in the needle-detected signals. The MUP occurrences were used as triggering sources to estimate their corresponding surface-detected MUPs (S-MUPs) using STA. The mean S-MUP size was calculated and divided into the maximum M-potential size to derive a MUNE. The MUNE values were consistent with those previously reported with other methods, and thenar MUNEs for the two trials were similar (249 +/- 78 and 246 +/- 90), with high test-retest reliability (r = 0.94, P < 0.05). DE-STA thus appears to be a valid and reliable method to obtain MUNEs. PMID- 15116374 TI - Proposed modification to data analysis for statistical motor unit number estimate. AB - Motor unit number estimation (MUNE) is an important electrophysiological technique for quantitative measurement of motor neuron loss. Although commonly used, there is no consensus concerning the optimal procedure for statistical MUNE, particularly regarding several operator-dependent variables. To assess the variables, we analyzed 500 sequential, submaximal compound muscle action potential (CMAP) responses at three or four stimulus intensities in 10 controls and 10 patients with amyotrophic lateral sclerosis (ALS). In both controls and ALS patients, we found that posttest filtering data based on 20% or 25% windows or 2, 2.5, or 3 SD excludes <5% of data. Windows of 10% or 15% excluded <5% of data in controls but not in ALS patients. Excluding data based on +/-2 SD, the coefficient of variation for final MUNE was 12% in controls and 6% in ALS patients. Group sizes of 30 or 50 and sample sizes of 300 to 500 sequential CMAP responses per run yielded the lowest coefficient of variation. We propose that statistical MUNE data should be analyzed based on excluding data >2 SD from the mean, because this is operator independent, includes the majority of data, effectively excludes clearly outlying data, such as fasciculations or movement artifact, and has a reasonable coefficient of variation. PMID- 15116375 TI - Continuous myofiber remodeling in uninjured extraocular myofibers: myonuclear turnover and evidence for apoptosis. AB - Unlike normal mature limb skeletal muscles, in which satellite cells are quiescent unless the muscle is injured, satellite cells in mammalian adult extraocular muscles (EOM) are chronically activated. This is evidenced by hepatocyte growth factor, the myogenic regulatory factor, Pax-7, and the cell cycle marker, Ki-67, localized to the satellite cell position using serial sections and the positional markers laminin and dystrophin. Bromodeoxyuridine (brdU) labeling combined with dystrophin immunostaining showed brdU-positive myonuclei, presumably the result of fusion of activated satellite cells into existing myofibers. One new myonucleus was added to every 1000 myofibers in cross section using a 12-hour brdU-labeling paradigm. The EOM thus appear to retain a stable nuclear population by an opposing process of apoptosis that results in myonuclear removal as visualized by terminal deoxynucleotidyltransferase-mediated nick end labeling (TUNEL). Activated caspase-3 was present in localized cytoplasmic domains extending from 10 to 210 microm within individual myofibers, suggesting segmental cytoplasmic reorganization. Understanding the cellular mechanisms that maintain this process of continuous myonuclear addition and removal in normal adult EOM may suggest new hypotheses to explain the preferential involvement or sparing of these muscles in skeletal muscle disease. PMID- 15116376 TI - Effect of aging on sensory nerve conduction study parameters. AB - Research evaluating the changes in nerve conduction with time has been limited to cross-sectional studies. We present a cohort study estimating how sensory nerve conduction study (NCS) parameters change with time when subjects are measured at two time-points. We evaluated 440 working adults by performing median and ulnar antidromic sensory NCS of both hands on two occasions, about 5.4 years (range, 4.3-7.0 years) apart. The rate of change in the NCS parameters was estimated using a mixed-models analysis controlling for each hand, gender, age, and body mass index (BMI). After controlling for gender, age, height, and BMI, the amplitudes of the median sensory nerve action potentials (SNAP) decreased by about 2.3 microV, peak latencies increased by 0.11 ms, onset latencies increased by 0.07 ms, and conduction velocities decreased by 1.1 m/s over 5 years. Corresponding values for the ulnar nerve were 1.75 microV, 0.06 ms, 0.04 ms, and 0.71 m/s, respectively. The findings are consistent with the findings of previous cross-sectional studies. The rate of change over time was not affected by hand (dominant versus nondominant hand), gender, age, or BMI at baseline. The rate of change seen with some of the median nerve parameters was significantly greater than that with the ulnar nerve. PMID- 15116377 TI - Novel dysferlin mutations and characteristic muscle atrophy in late-onset Miyoshi myopathy. AB - Miyoshi myopathy is characterized by weakness of the calf muscles during early adulthood. We report a case of late-onset Miyoshi myopathy presenting at 48 years of age, with novel mutations in the dysferlin gene. Muscle computed tomography clearly revealed severe atrophy in the soleus and medial gastrocnemius muscles. Even older patients with atrophy in the posterior compartment of the distal lower extremities and a relatively high serum creatine kinase level should be examined for the dysferlin gene. PMID- 15116378 TI - Compound muscle action potentials during repetitive nerve stimulation. AB - When using repetitive nerve stimulation to examine neuromuscular transmission, the change in compound muscle action potential (CMAP) size is usually assessed by measurement of negative-peak or peak-to-peak amplitude. Technological developments now allow automatic measurement of CMAP area, but some patients show increment of CMAP amplitude and decrement of CMAP area. This study systematically analyzed the changes in these CMAP parameters in 23 neurologically healthy subjects. CMAPs were recorded when the ulnar nerve was stimulated at frequencies of 1, 2, 5, 10, 20, 30, and 50 HZ (five pulses per train). CMAP amplitude showed significant increment within a train when stimulus frequency was above 5 HZ (probably due to increased muscle-fiber conduction velocity), whereas CMAP area hardly changed at any frequencies. Measurement of CMAP area produces less ambiguous results than amplitude measurement in repetitive nerve stimulation studies. PMID- 15116379 TI - Association of spinal and bulbar muscular atrophy with myotonic dystrophy type 1. AB - A man with spinal and bulbar muscular atrophy (SBMA) had a short (CTG)n expansion in the myotonic dystrophy protein kinase gene as well as (CAG)n expansion in the androgen receptor gene in leukocytes. The patient had the characteristic clinical findings of SBMA, but none of myotonic dystrophy type 1 (DM1). All of his three children (a son and two daughters) had the DM1 phenotype with long (CTG)n expansions. The daughters also had heterozygous long (CAG)n expansions. Postmortem examination of the patient revealed the characteristic pathological changes of SBMA as well as muscle degeneration compatible with DM1. Gene analysis of the organs disclosed unstable long expansions of the (CTG)n repeats, in contrast to the stable (CAG)n expansions. We have assumed that SBMA and DM1 developed independently in our patient, but cannot exclude the possibility that interactive gene effects increased somatic instability. PMID- 15116380 TI - Technology literature review: quantitative sensory testing. AB - The development of the personal computer has simplified the process of quantitating sensory thresholds using various testing algorithms. We reviewed the technical aspects and reproducibility of different methods to determine threshold for light touch-pressure, vibration, thermal, and pain stimuli. Clinical uses and limitations of quantitative sensory testing (QST) were also reviewed. QST is a reliable psychophysical test of large- and small-fiber sensory modalities. The results of QST are highly dependent on methodology and the full cooperation of the subject. QST has been shown to be reasonably reproducible over a period of days or weeks in normal subjects. The use of QST in research and patient care should be limited to instruments and their corresponding methodologies that have been shown to be reproducible. Literature data do not allow conclusions regarding the relative merits of individual QST instruments. PMID- 15116381 TI - Mycophenolate mofetil in dysimmune neuropathies: a preliminary study. PMID- 15116383 TI - Cytoarchitectonic and connectional organization of the ventral lateral geniculate nucleus in the cat. AB - The ventral lateral geniculate nucleus is a small extrageniculate visual structure that has a complex cytoarchitecture and diverse connections. In addition to small-celled medial and lateral divisions, we cytoarchitectonically defined a small-celled dorsal division. A large-celled intermediate division intercalated between the three small-celled divisions, which we divided into medial and lateral intermediate subdivisions. In WGA-HRP injection experiments, the different cytoarchitectonic divisions were shown to have connections with different nuclei. The medial division was reciprocally connected to the pretectum and projected to the superficial layers of the superior colliculus and the intralaminar nuclei. The medial intermediate division received projections from the intermediate layer of the superior colliculus and the lateral and interpositus posterior cerebellar nuclei, and projected to the intermediate layer of the superior colliculus, the periaqueductal gray of midbrain, and the intralaminar nuclei. The lateral intermediate divisions received projections from the pretectum, the intermediate layer of the superior colliculus, and the lateral and interpositus posterior cerebellar nuclei, and projected to the pretectum, superficial layers of the superior colliculus, and the pulvinar. The lateral division received projections from superficial layers of the superior colliculus and had reciprocal connections with the pretectum. The dorsal division received projections from the pretectum and had reciprocal connections with the periaqueductal gray of midbrain. The different cytoarchitectonic divisions of the ventral lateral geniculate nucleus are thus suggested to play different functional roles related to vision, eye and head movements, attention, and defensive reactions. PMID- 15116384 TI - Organization of connectivity of the rat presubiculum: I. Efferent projections to the medial entorhinal cortex. AB - The organization of the laminar and topographical projections from the presubiculum to the entorhinal area was studied in the rat by anterograde labeling with Phaseolus vulgaris leucoagglutinin and retrograde labeling with horseradish peroxidase conjugated to wheat germ agglutinin. We found that the pattern of presubiculo-entorhinal projections differs between the superficial and deep layers of the presubiculum. The superficial layers (layers II and III) of the presubiculum gave rise to bilateral projections to layers I-VI of the medial entorhinal area (MEA). Many terminals were distributed in layer III, fewer in layer II and the deep portion of layer I, and many fewer terminals in the deep layers (layers V and VI) of MEA. In contrast, the deep layers (layers V and VI) of the presubiculum gave rise to ipsilateral projections to the entorhinal area. Many axon terminals were distributed in layers V and VI of MEA and the most superficial portion of layer I of MEA, but very few in layers II and III. In addition, the ramifications in layer I extended to the lateral entorhinal area (LEA). Using two-dimensional unfolded maps of parahippocampal cortices, we elucidated the distinct topographical relationship in the presubiculo-entorhinal projection: 1) The septotemporal or longitudinal axis of the presubiculum corresponded to the axis on the MEA/LEA boundary, where the septal presubiculum projected toward the rhinal fissure and the temporal presubiculum projected away from the fissure. 2) The proximodistal axis of the presubiculum corresponded to the axis from the MEA/LEA boundary to the MEA/parasubiculum boundary that was virtually perpendicular to the MEA/LEA boundary, where the proximal portion of the presubiculum (close to the subiculum) projected to the region near the MEA/LEA boundary. PMID- 15116385 TI - Increase in motoneurons in the spinal nucleus of the bulbocavernosus of prepubertally castrated male Mongolian gerbils following delayed treatment with testosterone. AB - Sexual dimorphism in the spinal nucleus of the bulbocavernosus (SNB) of the Mongolian gerbil is achieved by two periods of postnatal increase, one in the first month after birth and one at puberty. The pubertal increase in motoneuron number is of particular interest because it occurs in a nearly adult animal. The purpose of this research was threefold. The first was to determine the response of the SNB in prepubertally castrated male gerbils receiving delayed hormone replacement as adults. Testosterone propionate (TP) treatment resulted in numbers of SNB motoneurons comparable to those seen in intact males, whereas androgen metabolites were less effective. The second purpose was to determine the latency of motoneurons to appear in response to TP. New SNB motoneurons appeared within 2 days of delayed TP replacement in prepubertally castrated males, and 16 days of treatment did not further increase SNB motoneuron numbers. The response of the motoneurons to TP appeared more rapid than the response of the bulbocavernosus (BC) muscle, scent gland, and seminal vesicles. The third purpose was to determine whether the new cells were connected to a target muscle. After 16 days of TP treatment, more motoneurons were labeled in the SNB following injection of a retrograde tract tracer into the BC muscle compared with the number seen in control animals. Thus, new motoneurons appeared in the SNB of prepubertally castrated male Mongolian gerbils within 2 days of the start of delayed TP treatment and were connected to a target within 16 days of TP treatment. PMID- 15116386 TI - Members of the Wnt, Fz, and Frp gene families expressed in postnatal mouse cerebral cortex. AB - The functions of Wingless-Int (Wnt) signaling, studied intensely in embryonic brain development, have been comparatively little investigated in the postnatal brain. We report remarkably patterned gene expression of Wnt signaling components in postnatal mouse cerebral cortex, lasting into young adulthood. Wnt genes are expressed in gene-specific regional and lamina patterns in each of the major subdivisions of the cerebral cortex: the olfactory bulb (OB), the hippocampal formation, and the neocortex. Genes encoding Frizzled (Fz) Wnt receptors, or secreted Frizzled-related proteins (sFrps), are also expressed in regional and lamina patterns. These findings suggest that Wnt signaling is active and regulated in the postnatal cortex and that different cortical cell populations have varying requirements for a Wnt signal. The OB, in particular, shows gene expression of a large variety of Wnt signaling components, making it a prime target for future functional studies. The penultimate components of the canonical Wnt pathway are the Tcf/Lef1 transcription factors, which regulate transcription of Wnt signaling target genes. Surprisingly, we found little Tcf/Lef1 expression in the postnatal neocortex. These observations suggest that noncanonical Wnt pathways predominate, which will require functional testing. However, Lef1 is widely expressed in the dorsal thalamus, and Wnt ligands and receptors are expressed, respectively, in cortical areas and thalamic nuclei that are interconnected. Thus, canonical Wnt signaling could be utilized in a major cortical input by Fz- and Lef1-expressing thalamic cells that innervate the Wnt expressing cortex. PMID- 15116387 TI - Expression pattern of lacZ reporter gene representing connexin36 in transgenic mice. AB - Targeted deletion of the connexin36 (Cx36) gene in the mouse genome leads to visual transmission defects, weakened synchrony of rhythmic inhibitory potentials in the neocortex, and disruption of gamma-frequency network oscillations. We have generated transgenic mice in which a reporter protein consisting of the exon1 coded N-terminal part of Cx36 fused to beta-galactosidase (N36-beta-gal) is expressed instead of Cx36. Here, we have used these mice for a detailed analysis of the reporter gene expression. By beta-gal staining of adult retina, we found expression of the lacZ reporter gene in the ganglion cell layer, in two rows of the inner nuclear layer, and in the photoreceptor layer. In the brain, beta-gal staining was present in gamma-aminobutyric acid (GABA)ergic neurons of the cerebellar nuclei, in non-GABAergic neurons of the inferior olive, in mitral cells of the olfactory bulb, and in parvalbumin-positive cells of the cerebral cortex. Outside the central nervous system, N36-beta-gal signals were detected in insulin producing beta-cells of the pancreas and in the medulla of the adrenal gland of adult Cx36(+/del[LacZ]) mice. This expression pattern suggests that Cx36 fulfills functional roles not only in several types of neurons in the retina and central nervous system but also in excitable cells of the pancreas and adrenal gland. PMID- 15116388 TI - A crustacean serotonin receptor: cloning and distribution in the thoracic ganglia of crayfish and freshwater prawn. AB - Serotonin (5-HT) is involved in regulating important aspects of behavior and a variety of systemic physiological functions in both vertebrates and invertebrates. These functions are mediated through binding to 5-HT receptors, of which approximately 13 have been characterized in mammals. In crustaceans, important model systems for the study of the neural basis of behaviors, 5-HT is also linked with higher-order behaviors, associated with different 5-HT receptors that have been identified at the physiological and pharmacological levels. However, no crustacean 5-HT receptors have been identified at the molecular level. We have cloned a putative 5-HT(1) receptor (5-HT(1crust)) from crayfish, prawn, and spiny lobster and have raised antibodies that recognize this protein in all three organisms. 5-HT(1crust) immunoreactivity (5-HT(1crust)ir) was observed surrounding the somata of specific groups of neurons and as punctate staining within the neuropil in all thoracic ganglia of crayfish and prawn. In the crayfish, 5-HT(1crust)ir was also found in boutons surrounding the first and second nerves of each ganglion and on the 5-HT cells of T1-4. In the prawn, 5 HT(1crust)ir was also found in axons that project across the ganglia and along the connectives. We found examples of colocalization of 5-HT(1crust) with 5-HT, consistent with the short-term modulatory role of 5-HT, as well as cases of serotonergic staining in the absence of a 5-HT(1crust) signal, which might imply that other 5-HT receptors are found at these locations. We also observed receptors that did not possess counterpart 5-HT staining, suggesting that these may also mediate long-term neurohormonal functions of serotonin. PMID- 15116389 TI - Expression of Trk receptors in the oculomotor system of the adult cat. AB - We examined the expression of the three Trk receptors for neurotrophins (TrkA, TrkB, and TrkC) in the extraocular motor nuclei of the adult cat by using antibodies directed against the full-Trk proteins in combination with horseradish peroxidase retrograde tracing. The three receptors were present in all neuronal populations investigated, including abducens motoneurons and internuclear neurons, medial rectus motoneurons of the oculomotor nucleus, and trochlear motoneurons. They were also present in the vestibular and prepositus hypoglossi nuclei. TrkA, TrkB, and TrkC immunopositive cells were found in similar percentages in the oculomotor and in the trochlear nuclei. In the abducens nucleus, however, a significantly higher percentage of cells expressed TrkB than the other two receptors, among both motoneurons (81.8%) and internuclear neurons (88.4%). The percentages obtained for the three Trk receptors in identified neuronal populations pointed to the colocalization of two or three receptors in a large number of cells. We used confocal microscopy to elucidate the subcellular location of Trk receptors. In this case, abducens motoneurons and internuclear neurons were identified with antibodies against choline acetyltransferase and calretinin, respectively. We found a different pattern of staining for each neurotrophin receptor, suggesting the possibility that each receptor and its cognate ligand may use a different route for cellular signaling. Therefore, the expression of Trk receptors in oculomotor, trochlear, and abducens motoneurons, as well as abducens internuclear neurons, suggests that their associated neurotrophins may exert an influence on the normal operation of the oculomotor circuitry. The presence of multiple Trk receptors on individual cells indicates that they likely act in concert with each other to regulate distinct functions. PMID- 15116390 TI - Postnatal development of dendritic spines on olfactory bulb granule cells in rats. AB - Postnatal morphological changes in granule cell dendritic spines and filopodia (collectively referred to as "spines/filopodia") were examined in the rat main olfactory bulb to characterize the development of the neural circuitry for olfaction. Granule cells were labeled with a membrane dye and confocal laser scanning microscope images of labeled spines/filopodia were acquired in the following three dendritic domains: apical dendrites in the external plexiform layer, those in the granule cell layer, and basal dendrites. In all three domains the proportion of typical spines slightly increased during development, with a concomitant decrease in the proportion of "stubby" spines lacking a neck; the proportion of filopodia remained unchanged, accounting for 20-40% of all protrusions. The mean diameter and length of the spine/filopodium population were nearly constant throughout development. On the other hand, the developmental pattern of the spine/filopodium density varied markedly, depending on the domain of the dendrites. In the external plexiform layer, the density did not change remarkably during development. The density in apical dendrites in the granule cell layer increased during the initial 2 postnatal weeks, then gradually decreased. The spine/filopodium density in basal dendrites, however, continued to increase until 4 weeks of age, and then began to decrease. These results suggest that a substantial amount of input-specific synaptic remodeling occurs in granule cells during development, which proceeds from superficial dendritic domains to deeper ones, occurring most prominently in the basal dendrites. PMID- 15116391 TI - Relationship between morphoelectrotonic properties of motoneuron dendrites and their trajectory. AB - The distribution and geometry of the dendritic trees of spinal motoneurons obey several well-established rules. Some of these rules are based on systematic relationships between quantitative geometrical features (e.g., total dendritic length) and the three-dimensional trajectory followed by dendrites from their origin to their termination. Because dendritic geometry partially determines the transmission of current and voltage signals generated by synapses on the dendritic tree, our goal was to compare the efficacy of signal transmission by dendritic trajectories that followed different directions. To achieve this goal, we constructed detailed compartmental models of the dendritic trees of three intracellularly stained biventer cervicis/complexus (BCCM) motoneurons and calculated the electronic properties of 361 dendritic paths. Each trajectory was classified according to its orientation, e.g., rostral, rostral-dorsal-lateral. The attenuation of current and voltage signals en route to the soma was strongly related to trajectory orientation. Trajectories with similar attenuation factors formed functional subunits that were arranged in distinct domains within the ventral horn. Changes in R(m) or R(i) had little effect on which trajectories belonged to each functional subunit. However, differences in the efficacy of signal transmission between subunits increased during high network activity (mimicked by decreases in R(m)). The most efficient subunit delivered two times more current and four times more voltage to the soma than the least efficient subunit. These results indicate that the input-output properties of motoneurons depend on the direction of the path taken by dendrites from their origin at the cell body to their terminals. PMID- 15116392 TI - Differential expression of Homer family proteins in the developing mouse brain. AB - Homer acts as a postsynaptic adaptor protein that links multiple targets, such as proteins involved in glutamate receptor signaling. We report the differential expression of the long form of Homer proteins produced from three distinctive genes during postnatal development of the mouse brain. Homer 1b/c and Cupidin/Homer 2a/b are widespread throughout the developing brain and are down regulated in hindbrain-origin regions, such as the cerebellum, pons, and medulla oblongata. In contrast, Homer 3a/b is restricted to the cerebellum, hippocampus, and neonatal olfactory bulb. In the cerebellum, Homer 1b/c and Cupidin/Homer 2a/b predominate in the postsynapses of developing granule cells, whereas Homer 3a/b is concentrated in the dendritic spines of Purkinje cells and their axons. The down-regulation of Homer 1b/c and Cupidin/Homer 2a/b is in marked contrast to the up-regulation of Homer 3a/b between the first and the second postnatal weeks. In the hippocampus, Homer 1b/c and Cupidin/Homer 2a/b are largely located in the CA1 region and the CA1-CA2 region, respectively, whereas Homer 3a/b is largely distributed in the CA2-CA3 region and peaks around the third postnatal week. In hippocampal cell cultures, Homer 1b/c and Cupidin/Homer 2a/b are expressed in inhibitory and excitatory neurons, whereas Homer 3a/b is largely expressed in excitatory neurons but not in inhibitory neurons. In the developing olfactory bulb, Homer 1b/c and Cupidin/Homer 2a/b are up-regulated in the granular, external plexiform, and glomerular layers, whereas Homer 3a/b drastically decreases in these regions within the first postnatal week. Cupidin/Homer 2a/b is also expressed in olfactory sensory neurons within a distinct olfactory epithelial zone and is then widely distributed to both the axons in the olfactory nerve layer and the cilia in the olfactory epithelium. These results demonstrate that Homer family members have distinct regional, cellular, and subcellular distributions in time and space during postnatal brain development. PMID- 15116394 TI - Estrogen receptor-beta in oxytocin and vasopressin neurons of the rat and human hypothalamus: Immunocytochemical and in situ hybridization studies. AB - Topographical distribution of estrogen receptor-beta (ER-beta)-synthesizing oxytocin (OT) and vasopressin (VP) neurons was studied in the hypothalamic paraventricular and supraoptic nuclei (PVH; SO) of ovariectomized rats. In distinct subregions, 45-98% of OT neurons and 88-99% of VP neurons exhibited ER beta immunoreactivity that was confined to cell nuclei. Neuronal populations differed markedly with respect to the intensity of the ER-beta signal. Magnocellular OT neurons in the PVH, SO, and accessory cell groups typically contained low levels of the ER-beta signal; in contrast, robust receptor labeling was displayed by OT cells in the ventral subdivision of medial parvicellular subnucleus and in the caudal PVH (dorsal subdivision of medial parvicellular subnucleus and lateral parvicellular subnucleus). Estrogen receptor-beta signal was generally more intense and present in higher proportions of magnocellular and parvicellular VP vs. OT neurons of similar topography. Immunocytochemical observations were confirmed via triple-label in situ hybridization, an approach combining use of digoxigenin-, fluorescein-, and 35S-labeled cRNA hybridization probes. Further, ER-beta mRNA was also detectable in corticotropin-releasing hormone neurons in the parvicellular PVH. Finally, double-label immunocytochemical analysis of human autopsy samples showed that subsets of OT and VP neurons also express ER-beta in the human. These neuroanatomical studies provide detailed information about the topographical distribution and cellular abundance of ER-beta within subsets of hypothalamic OT and VP neurons in the rat. The variable receptor content may indicate the differential responsiveness to estrogen in distinct OT and VP neuronal populations. In addition, a relevance of these findings to the human hypothalamus is suggested. PMID- 15116395 TI - FAD-linked sulfhydryl oxidase QSOX: topographic, cellular, and subcellular immunolocalization in adult rat central nervous system. AB - The distribution of the sulfhydryl oxidase QSOX in the rat brain was mapped using immunohistochemistry. QSOX is specifically expressed by neurons throughout the rostrocaudal extent of the brain as well as in the spinal cord. Although a majority of neurons express QSOX, different intensities of labeling were observed depending on the area: the strongest labeling was observed in the olfactory bulbs, isocortex, hippocampus, basal telencephalon, several thalamic and hypothalamic nuclei, cerebellum, and numerous brainstem nuclei. This study also describes the ultrastructural localization of QSOX in neuronal cells and demonstrates that the enzyme is associated with the Golgi apparatus. Finally, selected double immunohistochemistry showed that in the hypothalamus the highest levels of QSOX labeling were colocalized in neuron populations that express disulfide-bounded neuropeptides. These observations are consistent with a role of the enzyme in secreted peptide/protein folding. Data presented herein will serve as a basis for further investigations of the physiological function of QSOX in the central nervous system. PMID- 15116396 TI - Axon diameters and intradural trajectories of the dural innervation in the rat. AB - Neurophysiological studies have characterized the sensory responses of primary afferent nociceptors that innervate the intracranial dura. The present study used anatomical methods to examine in greater detail the axonal trajectories within the dura, as well as the axonal size distribution of the dural innervation. Immunostaining for CGRP in dural wholemounts revealed a network of fibers extending across the entire dura, with an especially dense plexus running along the borders of the transverse and superior sagittal sinuses. The plexus along the caudal border of the transverse sinus partially overlapped the dural area that shows the greatest density of mast cells. Visualization of axon bundles by DiI application in formalin-fixed tissue revealed two separate systems of fibers in the dura that could be distinguished by the orientation of their trajectories: one that runs parallel to the middle meningeal artery (MMA), and another with a more or less orthogonal orientation that runs rostromedially from the transverse sinus across the MMA. Axons traversed large distances across the dura, but the majority of the branching and arborization was usually concentrated in the distal part of the trajectory. In separate animals, measurement of myelinated axon diameters with electron microscopy showed that approximately one-third of the myelinated axons in the nerves supplying the dura (nervus spinosus and tentorial nerves) could be classified as A-beta, since they were comparable in size to the majority of axons in the trochlear nerve and the upper end of the size range in the trigeminal nerve (i.e., > 5 microm). PMID- 15116393 TI - Chemoarchitectonic subdivisions of the songbird septum and a comparative overview of septum chemical anatomy in jawed vertebrates. AB - Available data demonstrate that the avian septal region shares a number of social behavior functions and neurochemical features in common with mammals. However, the structural and functional subdivisions of the avian septum remain largely unexplored. In order to delineate chemoarchitectural zones of the avian septum, we prepared a large dataset of double-, triple-, and quadruple-labeled material in a variety of songbird species (finches and waxbills of the family Estrildidae and a limited number of emberizid sparrows) using antibodies against 10 neuropeptides and enzymes. Ten septal zones were identified that were placed into lateral, medial, caudocentral, and septohippocampal divisions, with the lateral and medial divisions each containing multiple zones. The distributions of numerous immunoreactive substances in the lateral septum closely match those of mammals (i.e., distributions of met-enkephalin, vasotocin, galanin, calcitonin gene-related peptide, tyrosine hydroxylase, vasoactive intestinal polypeptide, substance P, corticotropin-releasing factor, and neuropeptide Y), enabling detailed comparisons with numerous chemoarchitectonic zones of the mammalian lateral septum. Our septohippocampal and caudocentral divisions are topographically comparable to the mammalian septohippocampal and septofimbrial nuclei, respectively, although additional data will be required to establish homology. The present data also demonstrate the presence of a medial septal nucleus that is histochemically comparable to the medial septum of mammals. The avian medial septum is clearly defined by peptidergic markers and choline acetyltransferase immunoreactivity. These findings should provide a useful framework for functional and comparative studies, as they suggest that many features of the septum are highly conserved across vertebrate taxa. PMID- 15116398 TI - Is the songbird Area X striatal, pallidal, or both? An anatomical study. AB - Anatomical and neurophysiological studies have established that Area X, a songbird nucleus essential for vocal learning, is a basal ganglia structure, with mammalian striatal properties. However, Area X also sends a gamma-aminobutyric acid (GABA)ergic projection to the medial portion of the dorsolateral thalamus (DLM), a projection characteristic of the pallidum. These findings suggested that Area X contains both striatal and pallidal neurons. To test this hypothesis further, we investigated the neurochemistry and connectivity of Area X and its projections by using neurotransmitter antibodies, in combination with tracing studies. Like the mammalian striatum, Area X contains small enkephalin- and substance P-immunopositive neurons. Choline acetyltransferase-positive cells of Area X do not retrogradely label from DLM and are probably cholinergic interneurons similar to those in mammals. Like pallidal cells, large GABAergic cells project from Area X to the thalamus, but they also contain enkephalin, a characteristic of striatal neurons projecting to indirect pathway pallidal neurons. Moreover, many Area X cells are labeled with the pallidal marker Nkx2.1, but these do not include any thalamus-projecting neurons, suggesting that the projection cells are not of pallidal embryonic origin. Thus, although Area X combines both striatal and pallidal features, it is not a simple recapitulation of the mammalian circuit or of the avian lateral striatopallidal pathway: some individual Area X neurons may function as pallidal-like projection neurons but have striatal characteristics as well. Such heterogeneity of basal ganglia circuitry, both within and across species, may be facilitated by the developmental history of basal ganglia, which involves extensive migration and cellular intermixing. PMID- 15116399 TI - Optimization of bone-tissue engineering in goats. AB - Successful bone-tissue engineering (TE) has been reported for various strategies to combine cells with a porous scaffold. In particular, the period after seeding until implantation of the constructs may vary between hours and several weeks. Differences between these strategies can be reduced to (a) the presence of extracellular matrix, (b) the differentiation status of the cells, and (c) the presence of residual potentially immunogenic serum proteins. These parameters are investigated in two types of calcium phosphate scaffolds in a goat model of ectopic bone formation. Culture-expanded bone-marrow stromal cells from eight goats were seeded onto two types of hydroxyapatite granules: HA60/400 (60% porosity, 400-microm average pore size) and HA70/800. Scaffolds seeded with cells and control scaffolds were cultured for 6 days in medium containing autologous or semisynthetic serum, in the presence or absence of dexamethasone. Other scaffolds were seeded with cells just before implantation in medium with or without serum. All conditions were implanted autologously in the paraspinal muscles. After 12 weeks, bone had formed in 87% of all TE constructs, as demonstrated by histology. Histomorphometry indicated significantly more bone in the HA70/800 scaffolds. Furthermore, a significant advantage in bone formation was found when the constructs had been cultured for 6 days. In conclusion, both scaffold characteristics (porosity) and TE strategy (culturing of the constructs) were demonstrated to be important for bone TE. PMID- 15116400 TI - Mechanical strength of laser-welded cobalt-chromium alloy. AB - The purpose of this study was to investigate the effect of the output energy of laser welding and welding methods on the joint strength of cobalt-chromium (Co Cr) alloy. Two types of cast Co-Cr plates were prepared, and transverse sections were made at the center of the plate. The cut surfaces were butted against one another, and the joints welded with a laser-welding machine at several levels of output energy with the use of two methods. The fracture force required to break specimens was determined by means of tensile testing. For the 0.5-mm-thick specimens, the force required to break the 0.5-mm laser-welded specimens at currents of 270 and 300 A was not statistically different (p > 0.05) from the results for the nonwelded control specimens. The force required to break the 1.0 mm specimens double-welded at a current of 270 A was the highest value among the 1.0-mm laser-welded specimens. The results suggested that laser welding under the appropriate conditions improved the joint strength of cobalt- chromium alloy. PMID- 15116401 TI - Resistance to tearing of calf and ostrich pericardium: Influence of the type of suture material and the direction of the suture line. AB - The tearing of the valve leaflet of a cardiac bioprosthesis can cause early failure of this device, which is employed to replace a diseased native valve. This report involves the study of the behavior of 312 tissue samples (152 of calf pericardium and 160 of ostrich pericardium) treated with glutaraldehyde and subsequently subjected to tear testing. The samples were cut in the two principal directions: longitudinally, or root to apex, and transversely. They included a series of control samples that were left unsutured, and the remaining samples were repaired with the use of two different suture techniques: a running suture in the direction of the load and a telescoping suture perpendicular to the load. Four commercially available suture materials were employed: Pronova, nylon, Gore Tex, or silk. The unsutured control samples of both types of pericardium exhibited a similar anisotropic behavior in the tear test. The mean resistance to tearing of the calf pericardium was 24.29 kN m in samples cut longitudinally and 34.78 kN m in those cut transversely (p =.03); the values were 28.08 kN m and 37.12 kN m (p =.002), respectively, in ostrich pericardium. The series repaired with the telescoping suture always exhibited greater resistance to tearing, with values that ranged between 44.34 and 64.27 kN for the samples of calf pericardium and from 41.65 to 47.65 kN for those obtained from ostrich. These assays confirm the anisotropic behavior of calf and ostrich pericardium treated with glutaraldehyde when subjected to tear testing, as well as the loss of this behavior in ostrich pericardium after suturing. Suturing techniques, such as the telescoping model, that provide a greater resistance to tearing should be studied for use in the design of the valve leaflets of cardiac bioprostheses made of biological materials. PMID- 15116402 TI - The effect of dehydration history on PVA/PVP hydrogels for nucleus pulposus replacement. AB - The feasibility of the use of a copolymer gel prepared from blends of polyvinyl alcohol (PVA) and polyvinyl pyrrolidone (PVP) for endoscopic replacement of the nucleus pulposus of a lumbar intervertebral disc was examined in this study. Hydrogels were processed with the use of three freeze/thaw cycles to induce crystallinity. As-prepared samples were dehydrated to various levels: 70.4, 46.3, 25.1, and 10.3% of their as-prepared masses and subsequently rehydrated. The dehydration history controlled the dimensions upon swelling and caused distortion of the material, with major distortion occurring when the hydrogel reached around 25% of the initial hydrated mass. The dehydration history affected the mechanical behavior of the rehydrated gels. Increased dehydration resulted in increased compressive modulus for the reswollen gels. Experiments were performed to investigate the formation of a skin layer that was found on the hydrogels during the dehydration process. The skin was found to dehydrate quickly and form a barrier to further dehydration from the core. Rubber elasticity theory was used to describe the differences in the network characteristics between the skin and the core of a drying hydrogel. The dehydration/rehydration process used in this study and an implantation of a cadaver model demonstrate the feasibility of endoscopic nucleus replacement. PMID- 15116397 TI - Revised nomenclature for avian telencephalon and some related brainstem nuclei. AB - The standard nomenclature that has been used for many telencephalic and related brainstem structures in birds is based on flawed assumptions of homology to mammals. In particular, the outdated terminology implies that most of the avian telencephalon is a hypertrophied basal ganglia, when it is now clear that most of the avian telencephalon is neurochemically, hodologically, and functionally comparable to the mammalian neocortex, claustrum, and pallial amygdala (all of which derive from the pallial sector of the developing telencephalon). Recognizing that this promotes misunderstanding of the functional organization of avian brains and their evolutionary relationship to mammalian brains, avian brain specialists began discussions to rectify this problem, culminating in the Avian Brain Nomenclature Forum held at Duke University in July 2002, which approved a new terminology for avian telencephalon and some allied brainstem cell groups. Details of this new terminology are presented here, as is a rationale for each name change and evidence for any homologies implied by the new names. Revisions for the brainstem focused on vocal control, catecholaminergic, cholinergic, and basal ganglia-related nuclei. For example, the Forum recognized that the hypoglossal nucleus had been incorrectly identified as the nucleus intermedius in the Karten and Hodos (1967) pigeon brain atlas, and what was identified as the hypoglossal nucleus in that atlas should instead be called the supraspinal nucleus. The locus ceruleus of this and other avian atlases was noted to consist of a caudal noradrenergic part homologous to the mammalian locus coeruleus and a rostral region corresponding to the mammalian A8 dopaminergic cell group. The midbrain dopaminergic cell group in birds known as the nucleus tegmenti pedunculopontinus pars compacta was recognized as homologous to the mammalian substantia nigra pars compacta and was renamed accordingly; a group of gamma aminobutyric acid (GABA)ergic neurons at the lateral edge of this region was identified as homologous to the mammalian substantia nigra pars reticulata and was also renamed accordingly. A field of cholinergic neurons in the rostral avian hindbrain was named the nucleus pedunculopontinus tegmenti, whereas the anterior nucleus of the ansa lenticularis in the avian diencephalon was renamed the subthalamic nucleus, both for their evident mammalian homologues. For the basal (i.e., subpallial) telencephalon, the actual parts of the basal ganglia were given names reflecting their now evident homologues. For example, the lobus parolfactorius and paleostriatum augmentatum were acknowledged to make up the dorsal subdivision of the striatal part of the basal ganglia and were renamed as the medial and lateral striatum. The paleostriatum primitivum was recognized as homologous to the mammalian globus pallidus and renamed as such. Additionally, the rostroventral part of what was called the lobus parolfactorius was acknowledged as comparable to the mammalian nucleus accumbens, which, together with the olfactory tubercle, was noted to be part of the ventral striatum in birds. A ventral pallidum, a basal cholinergic cell group, and medial and lateral bed nuclei of the stria terminalis were also recognized. The dorsal (i.e., pallial) telencephalic regions that had been erroneously named to reflect presumed homology to striatal parts of mammalian basal ganglia were renamed as part of the pallium, using prefixes that retain most established abbreviations, to maintain continuity with the outdated nomenclature. We concluded, however, that one-to-one (i.e., discrete) homologies with mammals are still uncertain for most of the telencephalic pallium in birds and thus the new pallial terminology is largely devoid of assumptions of one-to-one homologies with mammals. The sectors of the hyperstriatum composing the Wulst (i.e., the hyperstriatum accessorium intermedium, and dorsale), the hyperstriatum ventrale, the neostriatum, and the archistriatum have been renamed (respectively) the hyperpallium (hypertrophied pallium), the mesopallium (middle pallium), the nidopallium (nest pallium), and the arcopallium (arched pallium). The posterior part of the archistriatum has been renamed the posterior pallial amygdala, the nucleus taeniae recognized as part of the avian amygdala, and a region inferior to the posterior paleostriatum primitivum included as a subpallial part of the avian amygdala. The names of some of the laminae and fiber tracts were also changed to reflect current understanding of the location of pallial and subpallial sectors of the avian telencephalon. Notably, the lamina medularis dorsalis has been renamed the pallial-subpallial lamina. We urge all to use this new terminology, because we believe it will promote better communication among neuroscientists. Further information is available at http://avianbrain.org PMID- 15116403 TI - Effect of slide track shape on the wear of ultra-high molecular weight polyethylene in a pin-on-disk wear simulation of total hip prosthesis. AB - Prosthetic joints appear to show a strong relationship between the type of relative motion and wear, requiring careful consideration in the design of wear simulators. This relationship was studied with a 12-station pin-on-disk device, specifically adapted for the wear simulation of prosthetic hip joints. Each station had a unique motion, characterized by the so-called slide track, the track of the pin on the disk. The slide track shapes included 10 ellipses, their aspect ratio (AR) varying from 1.1 to 11.0, and a circle and a straight line as extreme cases. Hence for the first time in hip wear simulation, the motion was systematically varied over a wide range. Conventional UHMWPE pins were tested against polished CoCr disks in diluted calf serum three times for 3 million cycles. Below the AR value of 5.5, the polyethylene wear factor and wear mechanisms agreed with clinical observations. Above this value, the wear factor decreased to unrealistically low values, and the wear surface topography differed from that of retrieved acetabular cups. The wear particles, however, were similar to those isolated from periprosthetic tissues, irrespective of the AR value. In conclusion, it is recommended that the AR value be kept well below the critical point of 5.5. PMID- 15116404 TI - Stripe wear rates in alumina THR--comparison of microseparation simulator study with retrieved implants. AB - Alumina-on-alumina hip implants with microseparation were run in a hip simulator for comparison of the nonseparation simulator mode and retrievals. The 28-, 32-, and 36-mm Biolox-forte implants were run to 5 million cycles with the use of 50% newborn calf serum. Howmedica Osteonics Trident cups with titanium backing were used in all sets. In standard (STD) and microseparation (MSX) mode, the typical biphasic wear trend was evident, but the MSX test mode had much higher magnitudes. There was a 5-fold increase for run-in wear and up to a 35-fold increase in steady-state wear. The stripe wear on the ball formed early, but did not progress in grade beyond 0.6 Mc. The locations of the stripes were similar in retrieved and simulator balls. However, the stripes from the simulator were narrower than short-term retrievals and much narrower than some long-term retrievals. The long-term retrieved balls had a grade of wear greater than the simulators. In vivo a broader range of motion occurs and this may lead to the wider stripe observed on the retrievals. These observations suggested that simulators could produce the loading and kinematics similar to a patient walking but not necessarily the variety of motions possible in the in vivo situation. PMID- 15116405 TI - Hierarchically biomimetic bone scaffold materials: nano-HA/collagen/PLA composite. AB - A bone scaffold material (nano-HA/ collagen/PLA composite) was developed by biomimetic synthesis. It shows some features of natural bone both in main composition and hierarchical microstructure. Nano-hydroxyapatite and collagen assembled into mineralized fibril. The three-dimensional porous scaffold materials mimic the microstructure of cancellous bone. Cell culture and animal model tests showed that the composite material is bioactive. The osteoblasts were separated from the neonatal rat calvaria. Osteoblasts adhered, spread, and proliferated throughout the pores of the scaffold material within a week. A 15-mm segmental defect model in the radius of the rabbit was used to evaluate the bone remodeling ability of the composite. Combined with 0.5 mg rhBMP-2, the material block was implanted into the defect. The segmental defect was integrated 12 weeks after surgery, and the implanted composite was partially substituted by new bone tissue. This scaffold composite has promise for the clinical repair of large bony defects according to the principles of bone tissue engineering. PMID- 15116406 TI - Long-term strength of ceramics for biomedical applications. AB - The influence of slow crack growth on the initiation of radial cracks at the lower surfaces of ceramic layers bonded to polymeric substrates is studied, with particular relevance to biomechanical systems, e.g., dental crowns and hip replacement prostheses. Critical loads are measured as a function of loading rate (dynamic fatigue) for model bilayers fabricated by epoxy-bonding selected clinical ceramics to polycarbonate bases. Radial crack initiation is observed in situ by viewing from below the transparent base during loading. Declines in the critical loads with diminishing load rate are consistent with slow crack growth of intrinsic flaws prior to radial crack pop in. A simple fracture mechanics relation incorporating a crack velocity function is used to analyze the data. Extrapolation beyond the data range enables long-lifetime (10 yr) estimates of sustainable loads. The procedure provides a basis for ranking ceramic types, and in particular for eliminating vulnerable candidate materials, for use in biomechanical systems. While slow crack growth is an important factor in failure, other mechanisms could operate in concert and even dominate under severe testing conditions, especially under cyclic loading. PMID- 15116407 TI - Absorbable microparticulate cation exchanger for immunotherapeutic delivery. AB - An absorbable microparticulate cation exchanger was synthesized as a versatile carrier for biologically active proteins. In this work, acid-terminated polyglycolide (or polyglycolic acid) microparticulates (PG-MP) were surface modified for either sustained release of cytokines or as a platform for immunomodulation. The intended goal was to achieve in situ recruitment/maturation of dendritic cells and activation of T cells for tumor immunotherapy. PG-MP were prepared with a volume weighted mean diameter of 7.02 micro (range: 2.09-14.58 micro). Accessible carboxylic acid groups were determined to be 0.3 mmol/g with a corresponding zeta potential of -21.87 mV in phosphate-buffered saline. Under low magnification, scanning electron microscopy (SEM) revealed a highly textured surface due to processing from repetitive jet milling. However, a moderately porous architecture was noted at higher magnification. Electron spectroscopy for chemical analysis was used to characterize the PG-MP surface before and after adsorption of human granulocyte-macrophage colony stimulating factor (GM-CSF). Adsorption of GM-CSF on PG-MP (PG-GMCSF) resulted in a modest increase in the surface atomic concentration of nitrogen (0.97%). Pretreating the surface with poly-L-lysine (PG/Lys-GMCSF) prior to adding GM-CSF produced a nearly threefold increase in the surface nitrogen concentration (4.20% compared to 1.47%). This manipulation not only increased loading content, but also prolonged the release of GM-CSF released from 6 days to 26 days. ESCA on the post-release PG-MP samples (PG-GMCSF and PG/Lys-GMCSF) revealed a similar residual surface nitrogen concentration (2.26% vs. 2.35%). The observation was consistent with irreversibly adsorbed GM-CSF. It is postulated that irreversibly bound GM-CSF is released over time as a function of microparticulate degradation. Biological activity of released GM-CSF was confirmed by the proliferation of a GM-CSF-dependent cell line (TF-1) in the presence of microparticulates. PG-MP mediated activation of T cells was achieved through irreversible adsorption of either antimouse cd3 plus antimouse cd28 monoclonal antibodies (alpha-cd3/cd28-MP) or antihuman CD3 plus antihuman CD28 monoclonal antibodies (alpha-CD3/CD28-MP) on PG-MP. Irreversibly adsorbed antibodies were capable of activating both resting mouse and human T cells. Intracellular flow cytometry on mouse T cells revealed that nearly 50% of the activated cells produced interferon-gamma (IFN-gamma). This was consistent with a TH-1 or cell-mediated response. In vivo efficacy was evaluated in a mouse flank tumor model showing a significant antitumor effect both alone and in combination. Combination therapy was most effective at preventing tumor implantation (8/8 mice) and was able induce tumor regression (4/7 mice) and/or stable disease (3/7 mice) in a regression model. In these studies, immunohistochemistry was used to confirm local recruitment of dendritic cells. In conclusion, the PG-MP represents a novel absorbable cation exchanger that can be readily manipulated to deliver biologically active proteins for immunotherapy. PMID- 15116408 TI - Properties of super stainless steels for orthodontic applications. AB - Orthodontic stainless-steel appliances are considered to be corrosion resistant, but localized corrosion can occur in the oral cavity. This study was undertaken to evaluate the properties of super stainless steels in orthodontic applications. Accordingly, the metallurgical properties, mechanical properties, corrosion resistance, amount of the released nickel, cytotoxicity, and characteristics of the passive film were investigated. Corrosion resistances of the specimens were high and in the following order: super austenitic stainless steel (SR-50A) > super ferritic stainless steel (SFSS) = super duplex stainless steel (SR-6DX) > 316L SS > super martensitic stainless steel (SR-3Mo) in artificial saliva, 37 degrees C. At 500 mV (SCE), current densities of SR-50A, SFSS, SR-6DX, 316L SS, and SR-3Mo were 5.96 microA/cm(2), 20.3 microA/cm(2), 31.9 microA/cm(2), 805 microA/cm(2), and 5.36 mA/cm(2), respectively. Open circuit potentials of SR-50A, 316L SS, SR-6DX, SR-3Mo, and SFSS were - 0.2, - 0.22, - 0.24, - 0.43, and - 0.46 V (SCE), respectively. SR-50A, SFSS, and SR-6DX released below 3 ng/ml nickel for 8 weeks, and increased a little with immersion time, and 316L SS released about 3.5 ng/ml nickel, but SR-3Mo released a large amount of nickel, which increased with immersion time. The study demonstrated that SR-50A, SR-6DX, and SFSS have high corrosion resistance and mild or no cytotoxicity, due to the passive film enhanced by synergistic effect of Mo + N or by high addition effect of Cr + W. All super stainless steels showed very low cytotoxicity regardless of their nickel contents, although SR-3Mo was found to be relatively cytotoxic. From these studies, these steels are considered suitable for orthodontic applications. PMID- 15116409 TI - The effect of cesium-containing leucite additions on the thermal and mechanical properties of two leucite-based porcelains. AB - It has been demonstrated that a Cs(2)O-stabilized leucite porcelain was susceptible to a stress-induced phase transformation, but toughening was not observed.1 It was hypothesized that if Cs(2)O-stabilized leucite core particles were added to a cesium-free matrix porcelain, selected or designed to enhance the toughening mechanism, toughening would occur. A commercial porcelain (VP) and a synthesized leucite-based porcelain (NP) were the matrix materials. Core particles of Cs(2)O (0.0-2.0 mol%) containing synthetic leucite were mixed with the two cesium-free matrix porcelains and vacuum fired into specimens for testing. The toughness of both types of matrix-based materials was dependent upon cesium content of the added core particles with a maximum toughness reached for those containing 0.75 mol% Cs(2)O. The toughness of the 0.75 mol% specimens (1.42 MNm(-3/2) for VP based and 2.15 MNm(-3/2) for NP based) was statistically (p <.02) higher than either of the matrix materials alone or the matrix materials containing added core particles of synthetic cesium-free leucite. However, the toughest materials (0.75 mol% Cs(2)O) were not the strongest materials, most likely because of large internal flaws. The results suggested that transformation toughening was possible. PMID- 15116410 TI - Migration of polyethylene particles around nonloosened cemented femoral components from a total hip arthroplasty-an autopsy study. AB - Three cemented femoral components from total hip arthroplasties retrieved at autopsy were examined in regard to the migration of birefringent polyethylene particles across the fixation interfaces. They were obtained from two patients who died from unrelated causes. They had been in place, respectively, for 8 and 9 years (first patient) and for 9 years (second patient). Osteolysis was present in the acetabular side only. The femoral components were not loosened. Histologic sections were performed at four levels of the femoral components, and polyethylene particles were counted with the use of a semiautomatic image analyzer. Sections below the tip of the stem were treated by deorganification and the lysate was centrifuged in the attempt to isolate wear debris. Particles were found at both interfaces (bone-cement and metal-cement) and in the cancellous bone between the trabeculae. They were also present in the Haversian canals of the cortical bone. Particles of various sizes were found in the medullary canal beneath the tip of the stem. In scanning electron microscopy, they appeared to have irregular shapes. Wear debris can migrate across the cement-bone interface of nonloosened implants. They appear to progress through the porosity of the cancellous bone. PMID- 15116411 TI - Chitosan membrane as a wound-healing dressing: characterization and clinical application. AB - Chitosan prepared from natural biopolymer chitin and cast into membranes has been tested as wound dressing at the skin-graft donor site in patients. Bactigras, a commonly used impregnated tulle gras bandage, served as a control. Chitosan membrane, prepared with a 75% degree of deacetylation and a thickness of 10 microm, was used in nonmesh or mesh form. The progress in wound healing was compared by clinical and histological examination. Itching and pain sensitivity of the wound dressed area was scored with the use of a visual analogue scale. Mesh chitosan membrane in contrast to the nonmesh membrane allowed blood to ooze into the surrounding gauze. After 10 days, the chitosan-dressed area had been healed more promptly as compared with the Bactigras dressed area. Moreover, the chitosan mesh membrane showed a positive effect on the re-epithelialization and the regeneration of the granular layer. The data confirm that chitosan mesh membrane is a potential substitute for human wound dressing. PMID- 15116412 TI - Comparative corrosion performance of black oxide, sandblasted, and fine-drawn nitinol wires in potentiodynamic and potentiostatic tests: effects of chemical etching and electropolishing. AB - The corrosion performance of sandblasted (SB) and smooth fine-drawn (FD) medical use nitinol wires was compared with the performance of wires with black oxide (BO) formed in air during their manufacture. Potentiodynamic and ASTM F746 potentiostatic tests in a 0.9 % NaCl solution were conducted on wires in their as received, chemically etched, aged in boiling water, and electropolished states. As-received wires with various surface finishes revealed breakdown potentials in the range from -100 mV to +500 mV; similar passive current density, 10(-6) A/cm(2); and a wide hysteresis on the reverse scan, demonstrating strong susceptibility to localized corrosion. Chemically etched wires with original black oxide displayed consistent corrosion performance and surpassed, in corrosion resistance, electropolished wires that showed significantly lower breakdown (400-700 mV) and localized corrosion potentials ( approximately -50 to +113 mV). Sandblasted and fine-drawn wires exhibited rather inconsistent corrosion behavior. In potentiodynamic tests these wires could perform with equal probability either on the level of pretreated BO wires or rather similar to as received wires. Both SB and FD wires revealed low breakdown potentials in the PS regime. SEM analysis performed before tests indicated that sandblasting was not efficient for the complete removal of the original scaling, and fine drawing aggravated the situation, resulting in a persistent scaling that contributed to the inferior corrosion performance. Inclusions (oxides, carbides, and oxidized carbides) inherited from the bulk and retained on electropolished surfaces are the cause of their inferior performance compared to chemically etched surfaces. In electropolished wires corrosion was initiated around inclusions. PMID- 15116413 TI - An experimental analogue to model the fibrous tissue layer in cemented hip replacements. AB - Fibrous tissue at the bone-cement interface of cemented joint replacements has been reported frequently in cases of revisions made necessary by aseptic loosening. This work describes the development of in vitro specimens suitable for biomechanical modeling of cemented femoral hip replacements with a fibrous tissue layer at the bone-cement interface. In particular, a series of uniaxial compression tests were performed on silicone elastomer specimens to identify a suitable analogue with similar mechanical characteristics to those reported for the fibrous tissue layer. A method was developed to apply the silicone elastomer at the bone-cement interface. This was examined for two types of cemented hip replacements implanted in composite femurs. The selected thickness of the elastomer layers was in the range of those found in clinical cases of aseptic loosening. Specimens produced by these methods could be used in preclinical biomechanical tests (such as stability or stress shielding tests) to assess the effects of a soft-tissue layer, to model in vitro a long-term-implant scenario, and to provide validation for similar finite element studies. PMID- 15116414 TI - Alpha-crystallin-incorporated collagen matrices as an aid for dermal wound healing. AB - This study evaluated the effects of noncovalently incorporated crystallin into the collagen matrix for dermal wound-healing processes in rats. Crystallin incorporated collagen matrix (CIC) showed better healing when compared to wounds treated with collagen matrix (CS) and without collagen (CR). Biochemical parameters and histological analysis revealed that increased wound contraction enhanced cell proliferation and efficient radical scavenging in the CIC group. The higher shrinkage temperature of CIC films when compared to CS groups suggested increased hydrothermal stability for the former material. An in vitro release study of CIC has showed sustained and time-dependent release of crystallin from the collagen matrix. These results demonstrate the possibility of using crystallin as therapeutic protein in the wound-healing process. PMID- 15116415 TI - Use and re-use of orthopedic implants in developing countries. PMID- 15116416 TI - Polydimethylsiloxane particles are not experimental in the human larynx. PMID- 15116417 TI - Dried-droplet probe preparation on AnchorChip targets for navigating the acquisition of matrix-assisted laser desorption/ionization time-of-flight spectra by fluorescence of matrix/analyte crystals. AB - We have developed a dried-droplet probe preparation method for peptide mass fingerprinting by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS), which uses AnchorChip targets and alpha-cyano-4 hydroxycinnamic acid (CHCA) as a matrix. Upon drying of a matrix and analyte mixture on the AnchorChip, salts and low molecular weight contaminants were pooled at the hydrophilic metal anchor, whereas 10-50 microm matrix/peptide crystals firmly adhered at the surface of a hydrophobic polymer and the entire target could be subsequently washed by submerging it in 5% formic acid for 2-3 min. Epifluorescence microscopy suggested that peptides were completely co localized with CHCA crystals at the AnchorChip surface. Fluorescent images of the probes were of good contrast and were background-free, compared with images taken by a video camera built into the ion source. CHCA/peptide crystals were easy to recognize at the surface and peptide mass maps were acquired from them without further adjustment of the position of the laser beam. These crystals were remarkably stable towards the laser depletion and almost no matrix-related ions were typically observed in the low m/z region of peptide mass maps. The sensitivity of the peptide mass mapping was at the low-femtomole level. PMID- 15116418 TI - A fragmentation study of a flavone triglycoside, kaempferol-3-O-robinoside-7-O rhamnoside. AB - A mass spectrometric method based on the combined use of electrospray ionization, collision-induced dissociation and tandem mass spectrometry has been applied to the structural characterization of the flavone triglycoside, robinin (3,5,7,4' tetrahydroxyflavone-3-O-robinoside-7-O-rhamnoside). The deprotonated molecule fragments by loss of the rhamnose glycan residue to yield the Y(7) (-) ion (m/z 593) and by scission of the robinose glycan residue to yield the radical anion [Y(3,0)-H](-.) (m/z 430). The Y(7) (-) ion fragments by scission of the robinose glycan residue to yield the radical anion of Y(7)[Y(3,0)-H](-.) (m/z 284). The [Y(3,0)-H](-.) radical anion fragments by loss of the rhamnose glycan residue to yield the radical anion Y(7)[Y(3,0)-H](-.) (m/z 284) and by scission to yield [Y(7)-H][Y(3,0)--H](-) (m/z 283). A fragmentation mechanism has been proposed. PMID- 15116419 TI - Mass spectra of nitro-beta,beta-dihalostyrenes. AB - The electron ionization (EI) fragmentation of nitro-beta,beta-dihalostyrenes depends strongly on the specific halogens present as well as on the position of the nitro group. Beta,beta-difluorostyrenes yielded mainly fluoroacetylene ions as the base peaks and o-nitro-beta,beta-dihalostyrene possibly furnished the ion with a structure similar to that of benzo[a]isoxazole (11). The structural identity of the ion was verified by comparing the collision-induced dissociation (CID) spectra of m/z 119 ions from p- and o-nitro-beta,beta-difluorostyrene, o nitro-beta,beta-dibromostyrene and compound 11. PMID- 15116420 TI - Detection of phenolic oxidation products in cider apple juice by high-performance liquid chromatography electrospray ionisation ion trap mass spectrometry. AB - Juice was prepared from cider apples of the cultivar "Kermerrien" under oxidative conditions. After isolation by solid-phase extraction, the phenolic fraction was subjected to high-performance liquid chromatography/electrospray ionisation mass spectrometry. SIM scans were performed at m/z values obtained in model solutions. The oxidation products, resulting from coupling between a molecule of caffeoylquinic acid and caffeoylquinic acid, catechin or dimeric flavan-3-ol, were detected. PMID- 15116421 TI - Multivariate approaches for efficient detection of potential metabolites from liquid chromatography/mass spectrometry data. AB - This work describes a novel method for rapid screening of unknown metabolites in urine samples that narrows down the list of potential metabolites. Prior to analysis by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS), urine samples were prepared using solid-phase extraction (SPE). Automatic curve resolution was used for deconvolution of the LC/MS data, followed by peak alignment. Preprocessed data were then used for metabolite pattern recognition using principal component analysis (PCA), parallel factor analysis (PARAFAC), and multilinear partial least squares (N-PLS). This approach enabled the rapid detection of metabolites of citalopram in urine by maximizing the information extracted. The metabolites thus identified were compared with earlier studies on the metabolism of citalopram. In addition, new, unreported metabolites were found and characterized by LC/MS/MS and accurate mass measurements. A combination of data from positive and negative ionization enhanced the identification of metabolites. PMID- 15116422 TI - Electron ionization mass spectral fragmentation of derivatized 4,5- and 5,6 epoxysterols. AB - The electron ionization (EI) mass spectral fragmentation of derivatized 4,5- and 5,6-epoxysterols was investigated. Interesting fragmentation processes involving a transannular cleavage of the epoxide ring after transfer of the trimethylsilyl group are significant in the case of 4,5-epoxysterol trimethylsilyl ethers (affording abundant fragment ions at m/z 403 and 404). Different pathways, which have been substantiated by deuterium labelling, are proposed in order to explain the formation of these ions. In contrast, this transfer is not significant in the case of 5,6-epoxysterol trimethylsilyl ethers. The EI mass spectra of these latter compounds appear to be very complex and to differ slightly according to the stereochemistry of the epoxy group. Acetate and trifluoroacetate derivatives of 4,5-epoxysterols display interesting EI mass spectra dominated by a fragment ion at m/z 332 resulting from cleavage of the steroid ring A. PMID- 15116423 TI - Tandem mass spectrometry of half-generation PAMAM dendrimer anions. AB - Ions derived from negative electrospray ionization of polyamidoamine (PAMAM) dendrimer generation 0.5 were subjected to ion trap tandem mass spectrometry. Ion/ion proton transfer reactions were used to manipulate the charge states of PAMAM precursor ions to form lower charge states from those initially formed by electrospray, as well as to facilitate the interpretation of the product ion mass spectra. Most of the products derived from dendrimer precursor ions could be rationalized by retro-Michael decomposition reactions. The dominant fragmentation channels are highly dependent on the composition of the counter-ions, which in this case are restricted to different numbers of sodium ions and protons, and whether the precursor ion is multiply charged or singly charged. An interpretation is given that is consistent with all of the observations made with the various anions associated with this study. The nature of the structural information that can be obtained via ion trap tandem mass spectrometry of the dendrimers is dependent on the types of precursor ions subjected to study. The tandem mass spectrometry data also provided information about the structure of faulty synthesis products present in the PAMAM dendrimer sample. PMID- 15116424 TI - A rapid and sensitive method for quantitation of nucleosides in human urine using liquid chromatography/mass spectrometry with direct urine injection. AB - Oxidized nucleosides are biochemical markers for tumors, aging, and neurodegenerative diseases. However, during the last decade, the analytical methods for nucleosides by gas chromatography/mass spectrometry (GC/MS) and high performance liquid chromatography (HPLC) with single-parameter detectors like electron-capture detection (ECD) have not been sufficiently rapid or reliable to detect nucleosides in urine and to analyze clinical samples. It has been reported (Dudley et al., Rapid Commun Mass Spectrom. 2000; 14: 1200) that liquid chromatography with electrospray mass spectrometry (LC/ESI-MS) is more specific and sensitive for analysis of nucleosides than HPLC with conventional detectors; however, this method required complex extraction steps. In the present work a direct LC/ESI-MS method for nucleosides without extraction of urine samples has been developed. Analysis of nucleosides using positive-ion mode with selected reaction monitoring effectively eliminated potential interferences from endogenous constituents of the urine. This highly selective and sensitive method made it possible to analyze urinary nucleosides with a lower limit of quantitation of 0.2 nmol/mL. The method has been validated, with both excellent linearity and reproducibility, in the calibration range from 0.2-400 nmol/mL. The correlation coefficients of the calibration curves were higher than 0.987. The coefficients of variation were in the range 0.03-14.92% (inter-day) and 0.54 14.39% (intra-day), respectively. PMID- 15116425 TI - A mass spectrometric and molecular orbital study of H2O loss from protonated tryptophan and oxidized tryptophan derivatives. AB - Protonated N-acetyltryptophan, oxindolylalanine (a mono-oxidized derivative of tryptophan), and N-acetyloxindolylalanine, as well as several di- and tripeptide derivatives containing oxindolylalanine, undergo a range of fragmentation reactions in the gas phase, including the loss of water. In order to elucidate the sites of water loss within these ions, and to determine the mechanisms associated with these processes, we have conducted a series of experiments employing multistage tandem mass spectrometry (MS/MS and MS(3)) in a quadrupole ion trap mass spectrometer, regiospecific structural labeling, and independent solution-phase syntheses of proposed product ion structures, coupled with the use of molecular orbital calculations at the B3LYP/6-31G* level of theory. We demonstrate that the loss of H(2)O from the amide carbonyl group of protonated N acetyltryptophan O-methyl ester occurs via a "side-chain-backbone" neighboring group reaction to yield a protonated carboline derivative. In contrast, the loss of water from the O-methyl ester of protonated oxindolylalanine results in the formation of a tricyclic structure by "backbone-side-chain" nucleophilic attack from the amino nitrogen to the C2 position of the indole ring. The O-methyl ester of protonated N-acetyloxindolylalanine was found to dissociate via the loss of water from both possible sites, i.e. from the side-chain indolyl oxygen and the backbone amide carbonyl group. An estimate of the relative preference for water loss from each site was obtained from the abundances of product ions formed from MS(3) analysis of regiospecifically labeled derivatives of N acetyloxindolylalanine, and from the results of molecular orbital calculations. These studies indicate the absence of a characteristic 'signature' ion or neutral loss for peptides containing oxindolylalanine residues under low-energy ion trap CID conditions. PMID- 15116426 TI - Enhancing capillary liquid chromatography/tandem mass spectrometry of biogenic amines by pre-column derivatization with 7-fluoro-4-nitrobenzoxadiazole. AB - This paper describes a capillary liquid chromatography/tandem mass spectrometry (LC/MS/MS) determination of biogenic amines enhanced by pre-column derivatization with 7-fluoro-4-nitrobenzoxadiazole (NBD-F). Biogenic amines including tryptamine, N-methylsalsolinol, histamine, and agmatine were studied. The biogenic NBD-amine derivatives could be quantitatively enriched in-line on 20 x 0.25 mm capillary columns packed in-house with 5 microm C(8) silica particles. In an electrospray ionization (ESI) source these derivatives were ionized effectively, and collision-induced dissociation (CID) produced predominant characteristic ions allowing sensitive MS/MS detection. Agmatine, a potential neurotransmitter/modulator, was taken as a reference compound to study the analytical figures of merit of the procedure. The detection limit of agmatine was estimated to be 0.6 ng/mL (signal-to-noise (S/N) = 3). A linear calibration curve in the range 15-1000 ng/mL agmatine with an r value of 0.9997 was obtained. Tissue samples of rat brain, stomach, and intestine were analyzed. Minimum sample pre-treatment was needed. Each analysis was accomplished within ca. 12 min. The concentration of agmatine was found to be 0.246, 3.31, and 0.058 microg/g wet tissue in the brain, stomach, and intestine, respectively. PMID- 15116427 TI - Effect of mutated transporters associated with antigen-processing 2 on characteristic major histocompatibility complex binding peptides: analysis using electrospray ionization tandem mass spectrometry. AB - A novel allele of transporters associated with the antigen-processing (TAP) 2 gene, TAP2*Bky2 (Val(577)), is significantly increased in Japanese patients with Sjogren's syndrome (SS), and has a strong association with SS-A/Ro autoantibody production in SS and autoantibody including anti-SS-A/Ro and anti-U1 RNP antibody in systemic lupus erythematosus (SLE). To determine the influence of this natural mutated TAP on peptides loaded onto MHC class I, we analyzed the repertoire of peptides loaded onto MHC class I on transfectants with TAP1 and TAP2 or mutated TAP2 by electrospray ionization tandem mass spectrometry (ESI-MS/MS). After comparison of the peptide profiles we identified three peptides from only mutated TAP transfectants. Moreover, one of these peptides is derived from snRNP A, which is a target for anti-U1 RNP antibody. To our knowledge this is the first report to show that the natural mutation of TAP2 changes the peptide profile loaded onto MHC class I molecules. PMID- 15116428 TI - Informed use of proteolytic inhibitors in biomarker discovery. PMID- 15116429 TI - Screening for novel essential genes of Saccharomyces cerevisiae involved in protein secretion. AB - We describe here a screening procedure devised for searching new genes involved in protein secretion in Saccharomyces cerevisiae. The screening procedure takes advantage of yeast strains constructed within the EUROFAN project, in which the promoters of the novel essential genes were replaced by the doxycycline-regulated tetO(7)-CYC1 promoter. This promoter is active in normal growth medium but results in downregulation of the gene in the presence of doxycycline. The yeast cells were grown in the presence or absence of doxycycline, and both the growth and secretion of the heat shock protein, Hsp150p, into the culture medium were determined. In seven strains there was a specific effect on protein secretion. In a strain in which the RPN5 gene was downregulated, the level of secreted Hsp150p was increased compared to the control culture. When RER2 was downregulated, cells secreted Hsp150p that was not of the mature size. In five strains, secretion was more severely reduced than cell growth. One of these downregulated genes, YGL098w, was recently reported to encode an ER-located t-SNARE, USE1. Four of the genes detected, NOG2, NOP15, RRP40 and SDA1, encode proteins involved in ribosome assembly, suggesting a possible new signalling pathway between ribosome biogenesis and production of secreted proteins. The results obtained here indicate that the present screen could be successfully used in larger scale to identify novel secretion-related genes. PMID- 15116430 TI - Functional analysis of the Candida albicans ALS1 gene product. AB - ALS1 encodes a cell surface protein that mediates adherence of Candida albicans to endothelial cells. The predicted Als1p has an N-terminal region, which contains a signal peptide; a middle region, which contains 20 36-amino acid tandem repeats; and a C-terminal region, which contains a glycosylphosphotidylinositol-anchorage sequence. We used site-directed mutagenesis to delineate the regions in Als1p required for endothelial cell adherence and cell surface expression of the protein. Mutant alleles of ALS1 containing either deletions or insertions were expressed in the normally non adherent Saccharomyces cerevisiae. These transformants were analysed for endothelial cell adherence and cell surface expression of Als1p. We found that mutations centred around amino acid 285 in the N-terminus completely abolished adherence, but had no effect on cell surface expression of Als1p. Deletion of 15 of the tandem repeats reduced adherence by 50%, whereas deletion of all abolished adherence completely, even though cell surface expression of the N-terminus of Als1p was maintained. Insertions into the C-terminus at amino acids 413 and 254 upstream of the stop codon resulted in a modest loss of adherence, while cell surface expression of Als1p was maintained. An insertion at amino acid 249 in the C-terminus caused complete loss of both adherence and cell surface expression, even though the glycosylphosphotidylinositol-anchorage sequence remained intact. These data suggest a model of Als1p in which the endothelial cell binding region is localized within its N-terminus, the tandem repeats are essential for the proper presentation of the binding site, and the C-terminus is required for localizing Als1p to the cell surface. PMID- 15116431 TI - Interaction of 6-phosphofructokinase with cytosolic proteins of Saccharomyces cerevisiae. AB - Hetero-octameric 6-phosphofructokinase (Pfk-1) from Saccharomyces cerevisiae is composed of two types of subunits, alpha and beta, which are encoded by the unlinked genes PFK1 and PFK2. Pfk single deletion mutants expressing only one type of subunit exhibit Pfk-1 activity in vivo which, however, is completely lost immediately after cell disruption. In order to elucidate the preconditions of the in vivo activity of the mutant enzymes composed of either alpha- or beta subunits, we have investigated their potential interaction with selected heat shock and cytoskeletal proteins, employing co-immunoprecipitation and immunofluorescence microscopy. Western blot analysis identified the mitochondrial chaperonin Hsp60, as well as the cytoskeleton proteins alpha-tubulin and actin, in complexes with Pfk-1 that were co-precipitated from a cell-free extract of a pfk2 single deletion mutant expressing only the alpha-subunit. The interaction of the corresponding mutant enzyme and Hsp60 was found to depend on the ATP concentration of the extract. Immunofluorescence microscopy displayed a conspicuously filamentous arrangement of the Pfk-1 mutant protein, exclusively in the pfk2 single deletion mutant. The analysis of structure and activity of Pfk-1 expressed in S. cerevisiae mutant strains defective in various heat shock proteins (TRiC/CCT, Hsp70, Hsp 104) and in the respective wild-type background did not reveal significant differences. PMID- 15116432 TI - Identification of genes encoding putative nucleoporins and transport factors in the fission yeast Schizosaccharomyces pombe: a deletion analysis. AB - In a systematic approach to study genes that are related to nucleocytoplasmic trafficking in the fission yeast Schizosaccharomyces pombe, the open reading frames (ORFs) of 26 putative nucleoporins and transport factors were deleted. Here we report the initial characterization of these deletion mutants. Of the 26 putative genes deleted, 14 were found to be essential for viability. Null mutations of essential genes resulted in failure to either complete one round or to sustain cell division. Four of the 14 essential genes, SPBC582.11c, SPBC17G9.04c, SPBC3B9.16c and SPCC162.08c, encode putative nucleoporins and a myosin-like protein with homologues NUP84, NUP85, NUP120 and MLP1, respectively, that are not required for viability in Saccharomyces cerevisiae, suggesting that their gene products perform critical functions in Sz. pombe. On the basis of combined drug sensitivity assays and genetic analysis we have identified five non essential null mutants that were hypersensitive to the microtubule depolymerizing drug thiabendazole (TBZ) and exhibited a cut phenotype upon TBZ treatment, suggesting possible involvement in microtubule function. Three of the corresponding ORFs, SPCC18B5.07c, nup40 and SPAC1805.04, encode putative nucleoporins with low similarity to the S. cerevisiae nucleoporins NUP2p, NUP53p and NUP133p, respectively. Further genetic analysis revealed that one of the nucleoporin genes, nup40, and another gene, SPCC1322.06, encoding a putative importin-beta/Cse1p superfamily protein may have a spindle checkpoint function. PMID- 15116433 TI - The KlSRB10 gene from Kluyveromyces lactis. AB - We report the cloning and sequencing of a gene from Kluyveromyces lactis with high homology to the SRB10 gene (alias UME5, SSN3, GIG2, NUT7, RYE5) from Saccharomyces cerevisiae and other organisms. The KlSRB10 gene is located in a similar configuration to that found in S. cerevisiae, flanked by NOT4 and a gene with high similarity to YPL041c. The translated protein contains 593 amino acids and the characteristic domains of kinases from the CMGC subgroup. The functional relationship to yeast SRB10 is demonstrated by complementation of mutant phenotypes in a haploid S. cerevisiae strain containing a null allele. PMID- 15116434 TI - Differential regulation by glucose and fructose of a gene encoding a specific fructose/H+ symporter in Saccharomyces sensu stricto yeasts. AB - Saccharomyces cerevisiae transports fructose through a facilitated diffusion system common to other hexoses and mediated by the Hxt proteins. The related species S. pastorianus (carlsbergensis) and S. bayanus produce, in addition, a specific fructose/H(+) symporter. We have previously cloned a gene (FSY1) encoding the active fructose symporter from S. pastorianus PYCC 4457. Expression of Fsy1p in a S. cerevisiae mutant (hxt-null) devoid of the facilitated diffusion system allows growth on fructose but not on glucose. Here we present results concerning the regulation of Fsy1p expression, both in S. pastorianus and S. bayanus, where it occurs naturally, and in suitably engineered S. cerevisiae transformants. To that purpose, we made use of both Northern blot analysis and a Fsy1p-GFP fusion protein. The expression of Fsy1p is strongly regulated by both the carbon source and its concentration in the growth medium. In S. pastorianus, as well as in S. bayanus, very low concentrations of either fructose or glucose induced expression but higher sugar concentrations prevented transcription of the gene. Glucose was considerably more effective than fructose in repressing FSY1 expression. Proper regulation of the gene in S. cerevisiae seems to be exquisitely dependent on sugar transport. Analysis of Fsy1 expression in S. cerevisiae mutants shows that repression is mainly dependent on Mig1p, the final effector of the main glucose repression pathway. Interestingly, Mig1p also seems to mediate repression of FSY1 expression by high maltose concentrations. PMID- 15116435 TI - Changes in Kaletra labelling. PMID- 15116436 TI - Rapid HIV test approved. PMID- 15116438 TI - Community setting challenges best practices. PMID- 15116439 TI - Know your boundaries in sexual assault litigation. AB - Four cases in which health care workers sexually assaulted patients show the "gray area" of hiring or supervisory negligence. PMID- 15116440 TI - What does IT have in store for nursing? AB - Globalization, our culture of change, and the information age shape 21st-century information technology. Find out the applications and implications for nursing. PMID- 15116441 TI - How to allocate the right staff mix across shifts, Part 2. AB - In Part 1, the author explained how to calculate, balance, and adjust your direct caregiver staffing needs. This month, read about how to include fixed staff in your budget for your unit's specific conditions. PMID- 15116442 TI - How do we withhold or withdraw life-sustaining therapy? AB - Nurse managers face many issues that concern withholding and withdrawing life sustaining therapy in acute care. The author offers a framework for clinical decision making, an overview of the legal and ethical principles involved, and strategies to enrich communication between patients, families, and the health care team. PMID- 15116443 TI - Managing pressure ulcers: a multisite CQI challenge. AB - Think preventing and tracking pressure ulcers is expensive? Treatment may cost more. The Nurse Executive Council of VHA Alabama partnered with KCI USA, Inc., to implement a CQI project to manage pressure ulcer incidence. PMID- 15116444 TI - Navigate a clinical pathway for uncomplicated MI patients. AB - At Naval Medical Center San Diego, a clinical pathway for uncomplicated myocardial infarction patients decreased length of stay, standardized practice, and significantly improved documentation and discharge practices. PMID- 15116445 TI - Expect truly unannounced surveys (and more) from the Joint Commission. AB - Prepare your unit for Joint Commission survey changes effective January 1, 2000. They come following an Office of the Inspector General investigation that found strengths and weaknesses in the survey process. PMID- 15116446 TI - Timely admits, transfers, and discharges. AB - Nurse managers implemented a critical care patient coordinator program that's improved patient flow, staffing, and problem identification. The hospital has increased the new department from 7 to 12 critical care nurses and applied the program to other units. PMID- 15116447 TI - When a domestic violence victim reaches out ... take her hand. AB - Health care professionals are often the first people domestic violence victims turn to for help, but many don't know how to respond. Be ready to embrace this educational opportunity with policies that incorporate the right screening, assessment, and interventions. PMID- 15116448 TI - Wade through wound care's healing technologies. AB - Tips to help you pick the right wound-care products, including dressings, prescription treatments, adjunctive therapies and products, and tissue load management products. PMID- 15116449 TI - Osteoradionecrosis, oral health and dental treatment. AB - Osteoradionecrosis (ORN) is a potentially debilitating complication of radiation therapy in the treatment of carcinoma in and around the oral cavity. Treatment of ORN is by surgical resection of all necrotic tissue and primary closure of the mucosa. This is supplemented with HBO given in 90-minute sessions at approximately 2.5 ATM of pure oxygen for 20 sessions prior to surgical intervention and followed by 10 additional postoperative dives. Antibiotics, such as penicillins or similar spectrum antibiotics, may be utilized preoperatively and for approximately one week postoperatively. PMID- 15116450 TI - Making your practice more productive. PMID- 15116451 TI - Professional liability for ADAA members. PMID- 15116452 TI - During economic uncertainty, patient comfort is king. PMID- 15116453 TI - Army's "look for xylitol first" program. AB - Xylitol is a sugar substitute not well known in the United States. This sugar substitute is not only low in calories but can also help prevent dental caries. The U.S. Army Dental Command's Health Promotion Program is constantly seeking additional prevention measures to enhance the oral health of America's Army. The Dental Command has created the "Look for Xylitol First" initiative aimed at training all members of the dental care team on the positive benefits of xylitol and to teach patients how to be smart consumers and evaluate products for their xylitol content. PMID- 15116454 TI - Atrial tachycardia ablation in a patient with double outlet right ventricle corrected by surgery. AB - The development of surgical and percutaneous techniques for treatment or palliation of congenital heart disease has prolonged survival in these patients and has increased late complications, particularly arrhythmias. Such arrhythmias are more frequently refractory to medical therapy, requiring percutaneous ablation. We present the clinical case of a 14-year-old child with complex congenital heart disease (double outlet right ventricle) who underwent two corrective surgeries (Rastelli operation and subsequent replacement of the homograft in the conduit connecting the right ventricle to the pulmonary artery; ventricular septal defect closure and tricuspid valve repair). After the second surgery the patient presented with wide complex syncopal tachycardia, refractory to medical therapy. Electrophysiologic study (EPS) identified an isthmus dependent atrial flutter that was successfully treated by radiofrequency (RF) ablation (a linear block was created along the cavo-tricuspid isthmus). Three months later a new episode of tachycardia occurred, but without syncope. The second EPS revealed an atrial tachycardia originating from the lateral wall of the right atrium, which was treated by ablation with focal application of RF energy. Four months after the last EPS the child remains free of arrhythmic symptoms, under no anti-arrhythmic therapy. PMID- 15116455 TI - Cardiac Doppler variation with volume status changes in general intensive care. AB - The authors studied the effect of volume status modification on cardiac Doppler features, with negative fluid balance and corresponding central venous pressure change. This was carried out in 64 patients admitted to the Intensive Care Unit, 24 of whom were under mechanical ventilation. With volume status change, the mitral E/A ratio showed a tendency to decrease, mitral E wave deceleration time decreased, isovolumic relaxation time increased, and the expiratory diameter of the inferior vena cava reduced and its inspiratory collapse increased. No significant correlation was observed between the parameters studied and volume changes, or between central venous pressure and fluid balance. Volume changes in critical care patients modify certain features of Doppler echocardiography, but the magnitude of such variations is unpredictable. PMID- 15116456 TI - Early detection of sympathetic myocardial denervation in patients with familial amyloid polyneuropathy type I. AB - INTRODUCTION: Type I familial amyloid polyneuropathy (FAP I) is an autosomal dominant inherited disorder due to a genetic defect in transthyretin and is characterized by deposition of amyloid in various organs and tissues. The principal manifestations are related to polyneuropathy and dysautonomia. The aim of this study was to assess cardiac involvement and to correlate the findings with neurological status. METHODS: 34 patients with FAP (15 male and 19 female; mean age 43 +/- 15 years) underwent I123-labeled metaiodobenzylguanidine (MIBG) myocardial scintigraphy in order to evaluate cardiac sympathetic innervation. In addition they underwent ambulatory blood pressure monitoring (ABPM) and two dimensional and Doppler echocardiography. Neurological involvement was quantified according to a neurophysiologic score (EMG; 0 = no abnormality and 100% = maximal disability). RESULTS: The mean value of cardiac MIBG uptake was 1.75 +/- 0.5 (normal = 2.6 +/- 0.3) and correlated inversely with the EMG score (r = -0.67; p = 0.001). In 27 (79%) of the 34 patients there was a decrease in MIBG accumulation, in 18 (53%) an alteration in the circadian BP pattern and/or an increase in systolic and/or diastolic BP loads at night, and in 17 (50%) left ventricular hypertrophy and/or diastolic dysfunction. Twenty-two patients were symptomatic and had a mean EMG score of 37.7 +/- 25% (group I). The remaining 12 were asymptomatic and without neurological involvement (group II). Group I was characterized by older age (48 +/- 15 vs. 33 +/- 10.2 years, p = 0.01), lower MIBG uptake (1.5 +/- 0.4 vs. 2.2 +/- 0.5, p = 0.001), higher systolic (129 +/- 16 vs. 119 +/- 6 mmHg, p = 0.01) and diastolic daytime BP (82 +/- 10 vs. 76 +/- 6 mmHg, p = 0.05), and higher systolic (119 +/- 17 vs. 105 +/- 7 mmHg, p = 0.01) and diastolic nocturnal BP (71 +/- 11 vs. 62 +/- 9 mmHg, p = 0.01) than patients in group II. In 21/22 patients in group I and in 6/12 in group II there was a decrease in cardiac MIBG activity. Sixteen patients in group I and 2 in group II had abnormal circadian BP pattern. Left ventricular hypertrophy was only seen in group I. CONCLUSIONS: Patients with FAP have a high incidence of cardiac denervation and an abnormal circadian BP pattern. These alterations in cardiac autonomic function precede the development of clinical manifestations and may be an important factor in determining the optimal timing for liver transplantation, which is currently the only way to control the progression of the disease. PMID- 15116457 TI - Brugada syndrome complicated with sudden death. AB - Brugada syndrome is an electrocardiographic diagnosis that is increasingly recognized as a cause of sudden cardiac death. The authors present a clinical case of a patient with a family history of sudden death, in whom a diagnosis of Brugada syndrome had been established, and who died suddenly. They also present a brief review of the main findings of this entity, particularly the diagnostic criteria and treatment of choice, since it is recognized that its prevalence will rise in the coming years. PMID- 15116458 TI - Double orifice mitral valve in an asymptomatic adult with an unusual combination of congenital malformations: a case report. AB - We report a case of an asymptomatic adult patient, with several congenital malformations including an infrequent variant of double orifice mitral valve, postductal aortic coarctation, bicuspid aortic valve and an aneurysm of the right Valsalva sinus. The loss of support of the right coronary cusp of the aortic valve caused major aortic regurgitation. With the exception of the mitral valve, which was left untouched because it was neither stenotic nor regurgitant, all the other abnormalities were successfully corrected, in a two-step surgical approach. PMID- 15116459 TI - Fibrinolysis in cardiac arrest. AB - Cardiac arrest is a classic contraindication for fibrinolysis, due to possible hemorrhagic complications. Most cardiac arrests, particularly those occurring out of-hospital, are caused by vascular thrombosis, including myocardial infarction and pulmonary embolism, in which fibrinolysis has proved to be an effective treatment. Indeed, there are several reports and clinical trials suggesting that fibrinolysis may be a safe and effective therapeutic option in patients with cardiac arrest of presumed cardiovascular cause. Based on a case of successful fibrinolytic treatment of a pulmonary embolism patient with cardiac arrest, these questions are reviewed. PMID- 15116461 TI - Congenital heart disease and pulmonary hypertension. PMID- 15116460 TI - Burden of disease from hypercholesterolemia in Portugal. AB - Cardiovascular diseases are the main cause of death in Portugal, in developed countries and indeed, worldwide. Hypercholesterolemia is a major risk factor for these diseases. What are the potential health gains to be obtained by reducing the prevalence of hypercholesterolemia in the Portuguese population? How are they to be estimated and quantified? Studies of the burden of disease aim to measure and evaluate the impact of a disease or group of diseases on overall levels of health. Although they do not strictly speaking constitute an economic evaluation, since no specific interventions are analyzed, burden of disease studies do provide an accurate picture of a specific health problem and its magnitude, as well as an indication of changes in health policy and in preventive or corrective measures that might lead to improvement. This paper reports the results of a study of the disease burden of hypercholesterolemia in Portugal, calibrated for data from the year 2000. In this study, the disease burden is estimated as the component attributable to hypercholesterolemia in DALYs (disability-adjusted life years). DALYs are a measure used by international organizations such as the World Health Organization and the World Bank. PMID- 15116462 TI - Neonatal cardiac surgery. PMID- 15116464 TI - Pulsed tissue Doppler: contributions of the study of patients with hypertrophic cardiomyopathy. PMID- 15116463 TI - Myocardial infarction in a patient with congenital coronary anomaly. AB - The authors describe the clinical case of a 38-year-old patient, with a history of smoking and hypercholesterolemia, who was admitted for non-Q wave acute myocardial infarction, and in whom coronary angiography revealed severe coronary disease and a congenital coronary anomaly. Unlike many congenital coronary anomalies that are manifested in ischemic disease, the nature of this patient's anomaly may have contributed to its benign clinical evolution, and influenced the therapeutic approach. PMID- 15116465 TI - Continuing education--what is all the fuss about? AB - Advances in medicine and technology are demanding increased, and more diverse, education and learning. To provide quality patient care perioperative nurses need to be up to date. Is it possible to accomplish this task in our era of healthcare staff shortages and budget cuts? Step outside the traditional classroom and see where it leads. Continuing education, or mandatory education hours are also requirements for a growing number of professional bodies. Can we continue to meet this demand? PMID- 15116466 TI - Leadership and mentoring. PMID- 15116467 TI - Giving effective presentations. AB - Apprehension about oral communication, or public speaking is rated as the number one fear among most individuals. Developing skill in, and comfort with, public speaking is important whether we are presenting oral reports and proposals, responding to questions, or training co-workers. Effective speakers are able to communicate information in a way that stimulates interest, helps the audience to understand and remember, and influences attitudes and behaviours. Many of us think that effective speakers are born rather than made. In truth most successful speakers work hard and invest a great deal of time and effort in to improving their speaking capabilities. Effective public speaking is a learned skill and activity that requires lots of practice. Like other learned skills, having a strategy with clear action steps can help you achieve your goal. PMID- 15116468 TI - Thinking outside the box: perioperative preceptorship. PMID- 15116469 TI - [Prevention and promotion in primary care consultations: priorities and effectiveness]. PMID- 15116470 TI - [Population survey to determine perceptions on preventive activities]. PMID- 15116471 TI - [Cardiovascular preventive activities in primary care]. PMID- 15116472 TI - [Recommendations on life style]. PMID- 15116473 TI - [Cancer prevention]. PMID- 15116474 TI - [Prevention of infectious diseases]. PMID- 15116475 TI - [Primary care prevention of mental health disorders]. PMID- 15116476 TI - [Preventive activities in the elderly]. PMID- 15116477 TI - [Prevention of unwanted pregnancy]. PMID- 15116478 TI - [Preventive activities in pregnancy]. PMID- 15116479 TI - [Preventive activities in climacteric]. PMID- 15116481 TI - Quo vadis CPD? PMID- 15116480 TI - [Prevention in childhood and adolescence]. PMID- 15116483 TI - Replacement of missing teeth with fixed prostheses. AB - This article explores the various treatment options available to replace missing teeth with a fixed prosthesis. It discusses the requirements for each treatment modality and its associated advantages and disadvantages. PMID- 15116482 TI - Preparing anterior teeth for indirect restorations. AB - Practitioners have many techniques available for the indirect restoration of anterior teeth. There is some confusion, however, as to what preparation technique is appropriate for each type of restoration, let alone which material to select for the production of the restoration. This article considers the different types of preparation recommended for veneers and resin-bonded, all ceramic and porcelain fused to metal crowns and gives an indication when each technique may be used. PMID- 15116484 TI - Management of a non-vital central incisor tooth with three root canals. AB - A macrodont permanent central incisor tooth with unusual root canal morphology became non-vital 18 months following trauma. Two root canals were initially identified and filled, but the patient continued to have symptoms and radiographic examination indicated apical periodontitis. Careful radiographic and clinical examination revealed a third root canal, which was subsequently treated, resulting in the resolution of symptoms and periapical healing. PMID- 15116485 TI - Evaluating the cost-effectiveness of orthodontic provision. AB - The assessment of orthodontic provision is important to determine if treatment was necessary and undertaken appropriately. The ICON objectively quantifies orthodontic treatment need, complexity and outcome and is a valuable occlusal index in the assessment effectiveness of orthodontic care. It is possible to develop cost-effectiveness models by analysing the costs and effectiveness of orthodontic treatment. Several methods are illustrated to compare the orthodontic provision of specialist orthodontists. PMID- 15116486 TI - Psychological aspects of bulimia and anorexia. AB - Basic information relating to eating disorders (EDs) will be presented. After first outlining incidence, mortality rates and diagnostic criteria, the actual experience of living with an ED will be explored. Finally, various casual theories will be discussed as well as treatment principles and strategies. PMID- 15116487 TI - MFGDP(UK): the Key Skills Portfolio. AB - The purpose of this paper is to enable general dental practitioners to understand how to compile an evidence-based portfolio for the MFGDP(UK) examination. The principles of evidence-based portfolios are outlined. There are six key skills that a candidate needs to understand in order to complete this portfolio. The six Key Skills are: medical emergencies in general practice, infection control, record keeping, radiography, legislation as applied to general practice and staff training. Specific advice is given on each key skill in order to complete the Portfolio. PMID- 15116488 TI - Rheumatoid arthritis of TMJ--a diagnostic dilemma? AB - Rheumatoid arthritis is a chronic disease affecting the musculo-skeletal connective tissue of the body with a strong predilection for the joints. The temporomandibular joint (TMJ) is commonly affected with rheumatoid arthritis in adults and children, but it is usually among the last joints to be affected. A case report is presented of a patient undergoing orthodontic treatment who developed unilateral condylar changes and whose serum was also positive for rheumatoid factor. The involvement of TMJ in several systemic disorders is a well established fact. When such disorders manifest in an atypical manner, it poses a diagnostic dilemma. This article aims to discuss various disorders causing condylar erosions. PMID- 15116489 TI - Case report: anterior lingual mandibular cortical bone concavity. AB - This paper presents a case of an anterior mandibular bone defect (synonym: Stafne's bone cavity, static bone cyst, lingual mandibular salivary gland depression) mistaken for periapical pathology and referred for treatment. PMID- 15116490 TI - Guidelines for the management of Sjogren's syndrome in secondary care. PMID- 15116491 TI - Revisiting the role of dentists in prescribing antibiotics. PMID- 15116492 TI - Clinical governance in primary dental care--getting started. PMID- 15116493 TI - Physical signs for the general dental practitioner. Case 13. Cushing's disease. PMID- 15116494 TI - Well-connected partnerships. PMID- 15116495 TI - A nurse-led service to identify and treat depression in primary care. AB - Setting up a GP mental health liaison service has led to clients at risk of depression being identified earlier, enabling them to access treatments in a primary care setting and easing pressure on other services. PMID- 15116496 TI - Ankylosing spondylitis: an overview. PMID- 15116498 TI - Acute or chronic? Pitfalls of using standard criteria to define wounds. AB - Specific terms such as infection, odour and exudate may be helpful for informing the course of patient care. However, classification of wounds as acute or chronic, although frequently used in practice, is a less useful way of characterising wounds. This paper challenges us to review the way we think about the types of wounds that are generally classified as either acute or chronic. PMID- 15116497 TI - Improving diabetes care through the NSF and GMS contract. AB - In response to the GMS Contract, which comes into effect this month, and the National Service Framework for Diabetes, one GP practice undertook an audit of patients with the condition. This revealed some gaps in the data recorded. The practice team worked to improve the information they gathered and entered on the computer system, and thus improve patient care in this area. PMID- 15116499 TI - The role of health professionals in supporting expert patients schemes. AB - A diagnosis of chronic disease can be devastating. Health-care professionals are one part of the team centred on supporting the person through that illness. Lay led-self-management programmes are another strand. This paper focuses on expert patients programmes and the importance of integrating self-management into current health-care provision. PMID- 15116500 TI - The value of thorough assessment in the management of cancer pain. AB - The number of patients likely to require management of cancer pain is increasing. However, although modern analgesia can help most patients achieve complete pain relief, some studies report widespread under-treatment of all types of pain. This paper describes approaches that health professionals can employ to ensure optimum pain management. PMID- 15116501 TI - Nurses and IT: a survey of use among community clinical staff. AB - A survey investigated the use of information technology among community nurses and other staff in one primary care trust. Provision of IT was poor across all the staff studied, with health visitors and community nurses lagging slightly behind others. The reasons for the low levels of use need further research so that barriers to using IT can be overcome. PMID- 15116502 TI - A nurse-surgical post cuts waiting times and extends nurses' skills base. AB - Government initiatives, including the reduction in junior doctors' hours, has resulted in nurses exploring new ways of expanding their roles to address unmet patient needs. This paper reports on the development of a nurse biopsy role in a dermatology department, which has not only extended the skills base of those involved, but also improved the care provided. PMID- 15116503 TI - Building a culture for safer systems: the experience of ambulance trusts. AB - This paper discusses the organisational process model--an approach that can be used to develop a flexible workplace culture to improve patient safety. Using the example of ambulance trusts and the aviation industry, the author considers the relationship between strategic management, line managers and the workforce--and how this affects patient safety. PMID- 15116504 TI - Latest guidelines on COPD: an overview. PMID- 15116505 TI - Using the benchmarking process to improve care after barium enema. AB - The benchmarking process in a multidisciplinary radiology department began by identifying the after-care needs of patients undergoing a barium enema. The benchmark focused on privacy and dignity and covered the related issues of nutrition, continence and bowel care. The resultant benchmark has changed practice and provided tools for ongoing assessment and audit. PMID- 15116506 TI - Understanding patients' beliefs and goals in medicine-taking. AB - The traditional paternalistic approach to health care ignores the individual needs of patients and results in many of them failing to take up treatment. Concordance, which centres on enabling patients to make informed decisions about all aspects of health care with the support of health professionals, is one approach increasingly being introduced in health-care settings. PMID- 15116507 TI - The importance of informed consent. PMID- 15116508 TI - Documentation in the dental office. PMID- 15116509 TI - Pennsylvania dental establishments in 2001. AB - Census Bureau reports for 2001 were used to develop data for "average" dental establishments in the counties of the state of Pennsylvania. There have been marked decreases in the number of residents per establishment and the proportional representation of smaller dental establishments (less than five employees) since 1980. Dental establishment "average" employee salaries ranged from $17,400 to more than $37,100 in the various counties. The need to monitor dental establishment activities beyond the confines of one's own facility is emphasized. PMID- 15116510 TI - How long are dental offices required to maintain hazardous and infectious waste records? PMID- 15116511 TI - Where Medicare goes ... the rest of the system may well follow CMS pay-for performance example. PMID- 15116512 TI - Consumers as directors. A revised, more active role for healthcare recipients may mean changes for the industry. PMID- 15116513 TI - Evidence-based medicine in 2006. A survey of health plan leaders identifies current and emerging strategies. PMID- 15116514 TI - Data mining. Stronger computer tools allow deeper analysis of medical research, patient care and insurance data. PMID- 15116515 TI - Robostaff. Some pharmacy and nursing tasks don't need the human touch. PMID- 15116516 TI - No alteration without authorization. Configuration management can select and arrange data as desired--while also protecting its content. PMID- 15116517 TI - Controlled medical vocabularies. A long-fallow field gets ready to bear important fruit. PMID- 15116518 TI - Remote access simplified. A large healthcare provider shares some practical lessons learned. PMID- 15116519 TI - Preventing surgical infections by keeping patients warmed. PMID- 15116520 TI - What can you say in checking references for new employees? PMID- 15116521 TI - Is spinal fusion surgery overused? PMID- 15116522 TI - Safe options for suction canister waste. PMID- 15116523 TI - Practical guide for improving performance. AB - Research has established that health care is not error free. The question facing perioperative units, as well as all health care services, is how to minimize the human factors that impact quality and safety. This two-part series on performance improvement in perioperative services was intended to help answer this question. Achieving performance excellence starts with a supportive work culture. The human issues of teamwork, communication, and leadership are crucial to achieving performance excellence. Next, perioperative caregivers must accept that all people make mistakes so systems and processes can be designed to be more "forgiving" of errors. Last, a planned and systematic approach must be used to measure, analyze, and improve performance. Successful implementation of performance improvement calls for strong partnerships between physicians, managers, and staff members. Performance excellence requires that everyone work together to ensure that perioperative care is safe, effective, appropriate, customer focused, and efficient. PMID- 15116524 TI - Fine-tuning your coding with modifiers. PMID- 15116525 TI - Good as (un)golden. A set of tactics and some understanding will help you get through to those difficult employees. PMID- 15116526 TI - Companion peace. This employee monitors residents' health, helps train staff and works for a small fee--and will never call in sick. PMID- 15116527 TI - The community of choice strategy. Ten steps to creating unmistakable value at your senior living facility. PMID- 15116528 TI - Dining done right. Keeping your facility full by revamping your dining service and menu. PMID- 15116529 TI - Improving employee efficiency and job satisfaction through intranets. PMID- 15116530 TI - The Internet's role in brand awareness. PMID- 15116531 TI - Allowing nonmembers into your member portal. PMID- 15116532 TI - Apple pie. PMID- 15116533 TI - Transparency builds trust. PMID- 15116534 TI - HIPAA processing woes. PMID- 15116535 TI - Workforce. Health care high. PMID- 15116536 TI - Patient safety. Making strides. PMID- 15116537 TI - Access & coverage. Common cause. PMID- 15116538 TI - Workforce. Here a nurse, there a nurse. PMID- 15116539 TI - Change your charges? PMID- 15116540 TI - End-of-life care. Keeping company. PMID- 15116541 TI - The data page. The cause + effect of medication errors. PMID- 15116542 TI - The last dot-com. An insider focuses on the front door of health care. PMID- 15116543 TI - How ready are we for reform? Without a shared vision, getting there will be difficult. AB - Providers, policy-makers and other stakeholders from across the political spectrum hope to harness election-year attention about the need for change into a true national discussion that yields a vision of what the American health care system should be and what strategies we need to take to make that vision a reality. PMID- 15116544 TI - Best practices and safety issues in the ICU. AB - Intensive care is the most complex and costly care provided in hospitals. These case studies look at what hospitals are doing to better staff their ICUs, prevent errors, incorporate information technology, involve family members and reduce unnecessary costs. PMID- 15116545 TI - Strategic savings. As supply costs climb, hospitals rethink their purchasing strategies. AB - Surging prices for supplies are intensifying the pressure on hospitals already reeling from skyrocketing liability, staffing and technology costs. The prices for certain supplies far outstrip general inflation and, executives say, could undermine the financial viability of some service lines. That's forcing some hospitals to rethink their purchasing strategies. PMID- 15116546 TI - 2003 Foster G. McGaw winner: Phoebe Putney Memorial Hospital, Albany GA. AB - By "doing the right thing and doing the smart thing," Foster McGaw Prize-winner Phoebe Putney Memorial Hospital in Albany, Ga., has made great strides in enhancing health care in a region that is among the 10 poorest congressional districts in the country. PMID- 15116547 TI - Report examines U.S. health behaviors by age, gender, race and other factors. PMID- 15116548 TI - Lack of financial incentives hinders Leapfrog efforts to improve patient safety. PMID- 15116551 TI - Despite obesity epidemic, children's efforts to lose weight cause for concern. PMID- 15116552 TI - A shared vision. PMID- 15116553 TI - Risk management. This is becoming a facilitywide activity. PMID- 15116554 TI - Middle Eastern nutrition. PMID- 15116555 TI - The technology trap. PMID- 15116556 TI - Heat-induced complex formation in solutions of alpha- and beta L-crystallins: a small-angle X-ray scattering study. PMID- 15116557 TI - The possibility of involvement of the pools of alpha-ketoglutaric acid in the biosynthesis of carotenoids in chloroplasts. PMID- 15116558 TI - Multiple deceleration of DNA synthesis during the S phase of cell cycle: study by flow cytometry method. PMID- 15116559 TI - The effect of melafen on the growth and energy processes in the plant cell. PMID- 15116560 TI - Conditions causing wavefront instability in a growing colony of bacterial cells with chemotactic activity. PMID- 15116561 TI - The modeling of the structure of the cysteine-rich domain of metabotropic glutamate receptors. PMID- 15116562 TI - Substitution of isoleucine M206 residue by histidine in the Rhodobacter sphaeroides reaction centers causes changes in the structure of the special bacteriochlorophyll pair molecule. PMID- 15116563 TI - Reciprocal dependence between pectinmethylesterase gene expression and tobamovirus reproduction effectiveness in Nicotiana benthamiana. PMID- 15116564 TI - A synthetic Au-rutin complex as a functional model of the active site of the Au protein from Micrococcus luteus. PMID- 15116565 TI - Oligonucleotides conjugated with acridine: a new type of fluorescence probes for DNA hybridization assay. PMID- 15116566 TI - Transgenic potato (Solanum tuberosum L.) plants expressing the gene of secretory nuclease from Serratia marcescens. PMID- 15116567 TI - Effect of natural and artificial defoliation on concentration and composition of extractive substances in leaves of birch (Betula pendula Roth.): methods of analysis and results. PMID- 15116568 TI - Molecular cloning and characteristics of allele variants (GATA)n, the microsatellite locus Du281 of parthenogenetic caucasian rock lizard (Darevskia unisexualis) genome. PMID- 15116569 TI - The study of the mechanism of binding of human ML1A melatonin receptor ligands using molecular modeling. PMID- 15116570 TI - Medium effect on the enzymatically triggered electron-exchange luminescence of the biologically important dioxetane. PMID- 15116571 TI - [Proper use of carbapenem antibiotics based on the feature in the treatment of infectious diseases (discussion)]]. PMID- 15116572 TI - [Antimicrobial prophylaxis in surgery]. AB - Antimicrobial prophylaxis is widely performed in any surgical procedures to prevent postoperative infections. However, we have neither double-blind placebo controlled studies nor sufficient surveillance of postoperative infections that are common in Europe and the United States, and therefore there is little convincing scientific basis accounting for the validity of this therapy. In addition, prophylactic agent is still uncovered by medical insurance despite the persistent arguments as to its necessity. To establish the guidelines in our own country, a greater deal of evidence needs to be accumulated. Strategies for antimicrobial prophylaxis should be determined based on the types of possible postoperative infections and the classifications of operations according to contamination levels in individual operative fields. This process may involve the precise selection of prophylactic agents for suspected contaminating bacterial species in each operative organ and their administration regimens suitable for the individual surgery. Upon selection of prophylactic agents for postoperative infections, various conditions should be considered: e.g., susceptibility, resistance, blood concentrations, urinary excretion, transition into body fluid and tissues, and adverse reactions. The first and second generations of cephem and cephamycin derivatives can be the first choice, but the use of various other antibacterial agents may be necessary for resistant bacterial strains such as methicillin-resistant Staphylococcus aureus (MRSA) and penicillin-resistant Streptococcus pneumoniae (PRSP). Cyclic therapy based on penicillins (including mixtures), cephems (including cephamycins) and phosphomycins also seems useful for such resistant strains. At present, there is only limited evidence supporting the importance of prophylactic agents. Controlled trials employing well-designed protocols that endure scientific criticism must be done with due consideration for medical economics. PMID- 15116573 TI - [Bacteria isolated from surgical infections and its susceptibilities to antimicrobial agents--special references to bacteria isolated between April 2002 and March 2003]. AB - Tendency of isolated bacteria from infections in general surgery during the period from April 2002 to March 2003 were investigated in a multicenter study in Japan, and the following results were obtained. In this series, 334 strains were isolated from 131 (75.3%) of 174 patients with surgical infections. One hundred and seventy-one strains were isolated from primary infections, and 163 strains were isolated from post-operative infections. From primary infections, anaerobic Gram-positive bacteria were predominant, while aerobic Gram-positive bacteria were predominant from postoperative infections. Among aerobic Gram-positive bacteria, although the isolation rate of Staphylococcus aureus was the highest, followed by that of Enterococcus faecalis from primary infections, the isolation rate of E. faecalis was the highest from postoperative infections. Among anaerobic Gram-positive bacteria, the isolation rate of Peptostreptococcus spp. was the highest from both types of infections. Among aerobic Gram-negative bacteria, Escherichia coli was the most predominantly isolated from primary infections, followed by Klebsiella pneumoniae and Pseudomonas aeruginosa in this order, and from postoperative infections, E. coli was the most predominantly isolated, followed by P. aeruginosa, Enterobacter cloacae, and Citobacter freundii. Among anaerobic Gram-negative bacteria, the isolation rate of Bacteroides fragilis group was the highest from both types of infections. The isolation rate of aerobic Gram-negative bacteria from primary infections and that of aerobic Gram-positive bacteria from postoperative infections were high in the last several years. We noticed no vancomycin-resistant Gram-positive cocci nor P. aeruginosa producing metallo-beta-lactamase. But we noticed cefazolin-resistant E. coli probably producing extended spectrum beta-lactamase. PMID- 15116575 TI - [Isolation-frequency and antibiotic susceptibility of Streptococcus pneumoniae, Haemophilus influenzae, and Pseudomonas aeruginosa isolated in regional core hospitals in the Nagaoka district of Niigata Prefecture (changes for 3 years)]. AB - We measured MICs of various antimicrobial agents against Streptococcus pneumoniae, Haemophilus influenzae, and Pseudomonas aeruginosa isolated in the Nagaoka district of Niigata Prefecture in 2000 (March-May), 2001 (January-May), and 2002 (March-May). S. pneumoniae: Fifty-six strains were isolated in 2000, 119 strains in 2001, and 88 strains in 2002. In 2000, 2001, and 2002, 24 strains (42.9%), 58 strains (48.7%), and 40 strains (45.5%), respectively, were penicillin-intermediate S. pneumoniae (PISP), and 4 strains (7.1%), 12 strains (10.1%), and 7 strains (8.0%), respectively, were penicillin-resistant S. pneumoniae (PRSP). Carbapenems had the most excellent antimicrobial activity, followed by penicillin G, against penicillin-susceptible S. pneumoniae (PSSP), PISP, and PRSP. H. influenzae: Seventy-six strains were isolated in 2000, 154 strains in 2001, and 91 strains in 2002. In 2000, 2001, and 2002, 6 strains (7.9%), 8 strains (5.2%), and 7 strains (7.7%), respectively, were beta-lactamase producing ampicillin (ABPC)-resistant strains (MIC > or = 2 micrograms/ml), showing no increase, and 14 strains (18.4%), 70 strains (45.5%), and 31 strains (34.1%), respectively, were beta-lactamase-non-producing ABPC-resistant strains (MIC > or = 2 micrograms/ml), showing a slight increase. Ceftriaxon, meropenem (MEPM), and levofloxacin had excellent antimicrobial activity against these resistant strains. P. aeruginosa: In 2000, 2001, and 2002, 135, 74, and 91 strains, respectively, were isolated, and 14 strains (10.4%), 17 strains (23.0%), and 24 strains (26.4%), respectively, were imipenem-resistant (MIC > or = 16 micrograms/ml), showing a slight increase. MEPM, biapenem, and ciprofloxacin had excellent antimicrobial activity against P. aeruginosa. PMID- 15116574 TI - [Nationwide surveillance of parenteral antibiotics containing meropenem activities against clinically isolated strains in 2002]. AB - The antibacterial activity of meropenem (MEPM) and other parenteral antibiotics against clinical isolates of 899 strains of Gram-positive bacteria, 1500 strains of Gram-negative bacteria, and 158 strains of anaerobic bacteria obtained from 28 medical institutions during 2002 was measured. The results were as follows; 1. MEPM was more active than other carbapenem antibiotics against Gram-negative bacteria, especially against enterobacteriaceae and Haemophilus influenzae. MIC90 of MEPM against Pseudomonas aeruginosa was the lowest of the drugs tested. MEPM showed low cross-resistant rate against both imipenem-resistant P. aeruginosa and ciprofloxacin-resistant P. aeruginosa. MEPM was active against most of the species tested in Gram-positive and anaerobic bacteria, except for multi-drug resistant strains including methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis (MRSE). 2. The proportion of extended-spectrum beta-lactamase (ESBL) strains was 3.1% (4 strains) in Escherichia coli and 1.9% (2 strains) in Klebsiella pneumoniae. Carbapenems including MEPM were active against these ESBL strains. In conclusion, the results from this surveillance study suggest that MEPM retains its potent and broad antibacterial activity and therefore is a clinically useful carbapenem; at present, 7 years after available for commercial use. PMID- 15116576 TI - [Transfer of cadrofloxacin, a novel fluoroquinolone antibacterial, into the gallbladder tissue and bile]. AB - Clinical pharmacological studies of a novel fluoroquinolone antibacterial drug, cadrofloxacin, were performed in 12 surgical patients by investigating the transfer of cadrofloxacin into the gallbladder tissue and bile. They were scheduled for cholecystectomy (n = 6) or had undergone a percutaneous transhepatic biliary drainage (n = 5) or gallbladder drainage (n = 1) at the Second Department of Surgery, Wakayama Medical University. A single dose of cadrofloxacin was orally administered to all patients at a dose of 200 mg. The results were as follows: 1) Serum and gallbladder tissue levels of cadrofloxacin after 2.9-5.7 hours were 0.6-2.5 micrograms/ml and 0.6-8.6 micrograms/ml, respectively. The gallbladder/serum ratios of drug concentration were 0.6-3.4. 2) Levels of cadrofloxacin in bile reached a peak of 2.5-12.9 micrograms/ml after individually different period of time. Cumulative bile recoverlies of cadrofloxacin (unchanged compound + glucuronide conjugate) within the first 6 hours were 0.02-0.49%, and 0.08-0.75% within 0-12 hours. The mean value of [glucuronide conjugate/unchanged compound] in bile was 37%, although the values depended on the patients. PMID- 15116577 TI - Concentration of clarithromycin and 14-R-hydroxy-clarithromycin in plasma of patients with Mycobacterium avium complex infection, before and after the addition of rifampicin. AB - Clarithromycin (CAM) and rifampicin (RFP) have both been recognized to be effective antibiotic agents against Mycobacterium avium complex (MAC) infection. Rifamycin derivatives including RFP and rifabutin modulate the CAM metabolism by inducing the hepatic cytochrome p-450 3A4. To clarify the effect of RFP on the CAM metabolism, we measured the plasma concentration of CAM and 14-R hydroxyclarithromycin (M-5), the major metabolite of CAM, in 9 patients suffering from MAC infection before and after the addition of RFP. After the addition of RFP, the mean plasma concentration of CAM significantly decreased, while that of M-5 did not. In addition, the amount of CAM + M-5 concentration also significantly decreased. As M-5 is less effective against MAC infection than CAM, more attention should thus be paid to the plasma CAM concentration in patients administered CAM and RFP concomitantly. PMID- 15116578 TI - [The effects of terrorist attacks on European Holocaust survivors compared to Greek Holocaust survivors]. AB - This research aims to examine whether there are differences in the level of anxiety, the perception of danger, the reliance on security forces and signs of psychological distress during times of war between two groups of Holocaust survivors: Jews of Greece and Jews of Eastern and Western Europe, while assessing their psychological ability to cope with the wave of terrorism against the Israeli population. The Jews of Greece have been portrayed as being different, both by the Nazis and by Jews from other countries, as well as by the Greeks themselves. Their strength of spirit, encouraging them to rebel and resist, as well as their physical strength, were renowned in the concentration camps. Each of these traits has been supported by specific documentation in history books dealing with the Jews of Greece during the Holocaust. Fifty-eight years after the Holocaust and before it disappears from historical record, we scientifically examined the psychological or mental capacity of these people to withstand the wave of terrorism. The study included 33 European Jewish Holocaust survivors and 38 Greek Jewish Holocaust survivors. The subjects completed four questionnaires. The main results are: 1. The average level of anxiety among Greek Holocaust survivors is clearly lower than that of other Holocaust survivors--an average of 10.00 compared to 16.48 (t = 4.83, p < 0.001). 2. The average level of psychological distress during times of war among Greek Holocaust survivors is 2.10 compared to 2.65 among other Holocaust survivors (t = 4.24, p < 0.001). 3. The average level of trust in the security forces among Greek Holocaust survivors is 3.67 compared to 2.70 among the other Holocaust survivors (t = 4.354, p < 0.001). 4. The average level of perception of danger among Greek Holocaust survivors is 2.75 compared to 3.39 among other Holocaust survivors (t = 2.60, p < 0.01). 5. The readiness to emigrate from Israel is 1.02 among Greek Holocaust survivors compared to 2.09 among other Holocaust survivors (t = 4.06, p < 0.001). The findings of the research support the theory that the Greek Holocaust survivors statistically demonstrate clear and more substantial psychological or mental immunity as compared to European Holocaust survivors. The differences between Greek Holocaust survivors and other Holocaust survivors increase when applied to a subgroup of former concentration camp prisoners. In conclusion, Holocaust survivors do not constitute a homogeneous group which responds uniformly to traumatic events. They are not to be regarded as one unit, since they bear different social and cultural burdens, as well as the universal values which they absorbed in their countries of origin. PMID- 15116579 TI - [What is the role of compulsory ECT therapy today?]. AB - Electro-convulsive therapy (ECT) has been effective for years, but it arouses opposition among patients and especially in the general public. ECT treatment is limited and compared to other medical treatment it is considered exceptional by the law, regulations and treatment personnel. A question arises as to the position of therapists regarding compulsory ECT treatment. A questionnaire was sent on this subject to all the units utilizing ECT in Israel. Opinions ranged from complete negation of compulsory ECT, to regarding such treatment as possible in cases when the patient is compulsorily hospitalized and/or when the patient's guardian supports this treatment. The authors' opinion is that the Law of Patients' Rights regarding special treatment when the patient is in extreme danger must be followed. The law requires that three physicians agree to the treatment, and compulsory treatment is no longer applicable when the danger passes. ECT treatment is important and imperative in certain conditions, especially conditions endangering patients' lives. In these conditions the law provides the authority to physicians to make decisions regarding treatment. PMID- 15116580 TI - [Intravenous leiomyomatosis]. AB - Intravenous leiomyomatosis is a rare tumor described only in women, in which benign smooth muscle cells grow into the pelvic venous channels. The tumor may cause life-threatening symptoms if there is involvement of the inferior vena cava or the right atrium. This is a case study of intravenous leiomyomatosis with left ovarian extension in a 42 years old woman. The patient was admitted for total abdominal hysterectomy due to uterus myomatosus and menorrhagia. During the operation a frozen section was sent for examination revealing intravenous leiomyomatosis. Thereupon, bilateral salpingo-oophorectomy was also performed. This article reviews the cause, disease, presentation, diagnosis, treatment and recurrence of intravenous leiomyomatosis. PMID- 15116581 TI - [Methemoglobinemia in children as a differential diagnosis of cyanosis]. AB - Methemoglobinemia is characterized by cyanosis with various degrees of severity. Symptoms range from asymptomatic to unconsciousness and death. Although cyanosis caused by methemoglobinemia is well-documented in the literature, it is rare and the consequences can be fatal. The cause of methemoglobinemia can be environmental, acquired, congenital or a combination of the above. Among the potential sources for methemoglobinemia are local anaesthetics that are in common use in hospitals and clinics, some of which can be purchased without prescription. Although these drugs are considered to be safe, they can still induce methemoglobinemia and can be life-threatening. In this review we describe patients admitted to the Pediatric Intensive Care Unit during the year 2000 suffering from cyanosis and diagnosed as having methemoglobinemia. PMID- 15116582 TI - [The impact of the annual scientific meetings of the Israel Society of Rheumatology as measured by publication rates of the abstracts in peer-reviewed journals]. AB - OBJECTIVES: We aimed to examine the impact and quality of the research presented in the Israel Society of Rheumatology (ISR) annual scientific meetings by measuring publication rates of the abstracts in peer-reviewed journals and investigating the factors that influenced publication. METHODS: We examined the outcome of all 79 abstracts submitted to the ISR for the 1998-2000 annual meetings. A MEDLINE search of all abstracts, by authors, topics and keywords was performed. Senior authors of abstracts not found to be published in this search were interviewed regarding publication and factors influencing submission. We described the effect of variable factors on the rate of publication. RESULTS: As of September 2002, 63 (80%) abstracts were published in peer-reviewed journals or are currently in-press. Most abstracts were published in prominent journals (with a high impact factor). The majority of the abstracts (61%) were published in rheumatologic journals, 65% of the studies originated from tertiary centers and 19% of the studies were multicenter. The most common diseases studied were antiphospholipid syndrome (20%), systemic lupus erythematosus (19%) and inflammatory arthritis (18%). Most of the studies were of disease pathogenesis (35%) and clinical manifestations (33%). The most common study designs were basic science (34%). An overall 57% of the studies reported "positive" results and 9% reported "negative" results. None of the factors studied were associated with publication or non-publication. The main cause cited by authors for not publishing their abstract was lack of time to prepare a full paper or a desire to further expand the study. Within this group of 16 authors of abstracts, 11 authors still plan to submit a paper. CONCLUSION: The ISR annual meetings have an important clinical scientific impact as measured by the high rate of abstracts published as full length articles in leading peer-reviewed journals. PMID- 15116583 TI - [A new definition for acute myocardial infarction]. AB - The recently introduced cardiac troponins are highly sensitive and specific markers for myocardial cell necrosis. The availability of these markers required a change in the classical definition of acute myocardial infarction. This definition of acute myocardial infarction, issued in 2000 by the American College of Cardiology and the European Society of Cardiology, requires elevation of cardiac biomarkers (preferably troponins). The new definition has multiple medical, epidemiological, social, occupational and other implications. As many as 30% of patients previously diagnosed with unstable angina now receive the diagnosis of myocardial infarction. These additional patients change the epidemiology of acute myocardial infarction. New approaches to issues of insurance and employment following myocardial infarction are necessary following the introduction of the new definition. A few aspects of the new definition of myocardial infarction remain controversial, especially those regarding mild myocardial damage following percutaneous coronary intervention. The Israel Heart Society has recently joined other such Societies in the world and endorsed the new definition. While some further refinement of the definition is probably required, we believe it is the best tool available today for the working diagnosis of myocardial infarction and call upon physicians to learn it and use it on a routine basis. PMID- 15116584 TI - [Morbidity in the concentration camps of the Third Reich]. AB - Life in the concentration camps of the Third Reich was like living on another planet. The prisoners, stripped of all rights, experienced constant humiliation, uncertain survival and endless terror. Living conditions were harsh, characterized by crowding, poor sanitation and personal hygiene, lack of proper clothing and heating. The days began early with long marches and slave labor. Sleep was short and interrupted, and fatigue was constant and severe. Above all hoovered the dark cloud of ever-present famine. The prisoners were given about a fourth of the daily calorie requirements, and the food lacked vital components such as vitamins and other essential ingredients. The psychological stress was extreme, yet morbidity and mortality were mainly due to infections, injuries and hunger. Lice, scabies and other skin diseases were common. Typhus fever was ever present, both endemic and epidemic, with a fatal outcome. Many suffered from tuberculosis, typhoid, dysentery, pneumonia and other infections diseases. Injuries were common, caused by beating, punitive whiplashing and other forms of physical abuse, gunshot wounds and dog-bites. Skull injuries with brain contusions and hemorrhages were prevalent, as well as fractured limbs, ribs and pelvic bones. Blunt injuries to chest and abdomen often had fatal outcomes due to the perforation of viscera and peritonitis or as a result of massive hemorrhage from ruptured blood vessels. The harsh winters were marked by frozen gangrenous limbs and hypothermia. Yet, the most ominous condition was the "hunger disease" with its multiple clinical expressions which, in their extreme form, led to the emaciated "musleman" and eventual death. PMID- 15116585 TI - [Changing the therapeutic approach to acute otitis media in children]. AB - Acute Otitis Media (AOM) is the most common reason for pediatrician's visits and for antibiotic prescription in childhood. A significant rise in bacterial resistance to antibiotic treatment has been detected in recent years. Accordingly, the attitude towards antibiotic treatment for AOM has been re evaluated. Due to various difficulties in ear examination, physicians overdiagnosis Otitis Media with Effusion (OME) as AOM, leading to unnecessary prescription of antibiotics. The natural history of AOM shows spontaneous improvement without complications. Studies that have examined antibiotic treatment versus placebo in AOM have shown only minimal advantage for the antibiotic therapy in symptom reduction. Critical appraisal of the literature according to Evidence-based Medicine (EBM) criteria has led to several meta analyses that showed only a minor advantage for antibiotics over placebo in AOM. In the Netherlands, the approach to AOM is that of delayed prescribing: symptomatic therapy is given for the first 24-72 hours and an antibiotic drug is prescribed only if symptoms persist after this initial period. This review examines the difficulties in reaching an accurate diagnosis of AOM and describes the natural history of AOM and evaluates the studies and meta-analyses comparing antibiotics to placebo. The Dutch approach to AOM will be discussed as an option and a recommended basis for reduction in antibiotic prescriptions for AOM. PMID- 15116586 TI - [Primary care approach to children with special needs]. AB - Children with special needs have medical, educational, social and emotional needs which are significantly greater than the average needs of their healthy peers. They and their families have to struggle with the physical, economical and emotional burden imposed on them by their special needs. The role of primary physicians is to lead a team that is capable of diagnosing, treating and coordinating treatment and support for the child and the family. Case management and anticipatory guidance are the cornerstone of the art of dealing with children who have special needs. Primary community physicians are expected to conduct a survey in order to identify the children with special needs in the community and to prepare an intervention plan for each of them. Early detection and proper management of the problems are expected to help these children to maximize their potential and integration in society. PMID- 15116587 TI - [A psychosocial view of a number of Jewish mourning rituals during normal and pathological grief]. AB - This article describes the three stages of normal and pathological mourning, emphasizing the constellation embodied in Judaism for this process. These stages are: shock, acute mourning, working through and reconciliation. We present the important question: "How to define pathological mourning?" It is certainly not only a matter of extending beyond the accepted time limits of the mourning process, but also a question of the intensity of mourning in ones daily life, the degree of being preoccupied with it, and the degree of priority that this mourning process has in an individual's life. A number of forms of pathological mourning, during the three mentioned stages, are described, with special attention to Jewish mourning rituals, especially: The "rending of the garments" (Kriyah), the Kaddish, the Shiva, and the termination of mourning after a fixed period of time. One of the possible interpretations of these rituals is that they prevent and neutralize manifestations of aggression and violence. This is an analogue to the function of biological (genetic) rituals which according to the theory of Konrad Lorenz, also minimize the dangerous aggression between the species in nature. The religious ritual converts an aggressive behavior to a minimal and symbolic action, often re-directed, so that an originally dangerous behavior becomes a ritual with an important communicative function. PMID- 15116588 TI - [The geriatric department in a general hospital]. AB - Geriatric medicine encompasses different aspects including geriatric evaluation, geriatric rehabilitation, chronic institutional care and acute geriatric care. In the last few years, the place of the geriatric department in the realm of the general hospital has come under discussion. In Israel, most general hospitals have a geriatric department, but its characteristics vary from one hospital to another. It would seem that a geriatric department in a general hospital is obligatory, but it can only be complementary to the internal medicine department, not replace it. The geriatric department should provide services for elderly patients with special needs such as multi-faceted illness, acute illness combined with a loss in general and cognitive function and facilitate services for elderly patients with expectations of extended stay and/or chronic hospitalization, etc. In addition, integration should be encouraged between the department and chronic care institutions and it should act as an "umbrella consulting firm" to the different hospital departments that provide treatment for most of the patients that are 65 years old and above. Among the important functions of the geriatric department is the training of interns and the teaching of geriatrics in nursing schools. The existence of geriatric departments in general hospitals will help in preserving the status of internal medicine. PMID- 15116589 TI - [Waiver of medical confidentiality]. AB - The duty to maintain medical confidentiality is one of the fundamental and well known obligations imposed on physicians, and is incorporated in the Law of Patient's Rights 1996 (the Law). Medical confidentiality applies to information conveyed to a physician for medical purposes trusting that the physician will not disclose it. The patient's identity and financial arrangements are not confidential. Any physician who breaches this duty may face civil, disciplinary or criminal lawsuits. The Law, as well as specific provisions in other rulings, defines the exceptions to the duty of upholding medical confidentiality. According to these provisions there are special circumstances in which a physician may, and even should, transfer medical data relating to his/her patients. One of these exclusions is the patient's consent to reveal the information. This article presents the legal infrastructure of medical confidentiality, and the means and circumstances in which medical confidentiality can be waived. Waiver of medical confidentiality does not constitute permission to convey all the information to one and all, and the Law determines that only the data required for specific purposes should be transferred. The way by which the patient waives his right for confidentiality is not defined by the Law or by regulations. A patient's consent to waive his right for confidentiality can be allegedly obtained in writing, orally or by behavior. It seems that the appropriate manner of waiving medical confidentiality is in writing. The waiver form should specify the nature and extent of information that can be conveyed, and it must be signed by the patient. The doctor is also responsible to ensure privacy during the transmission of the data. Specific subjects which are associated with the duty to maintain medical confidentiality are also discussed in this paper. In cases when a man's life or safety is at stake, the duty of a physician to protect life overweighs his duty to entrusted confidentiality. In such cases, the physician should transfer the information even if the patient did not waive his right for secrecy. This obligation also applies to patients who suffer from specific diseases, for example HIV. The duty to maintain medical confidentiality is relevant to the patient's relatives. Obtaining written consent to convey the information to family members is not practical. However, it is recommended that a physician should provide the information only to relatives he knows, and only when he can reasonably assume that the patient will agree. PMID- 15116590 TI - [Ivan Illich's revenge and death: on iatrogenesis and related phenomena]. AB - Dr Ivan Illich, the devoted fighter for the elimination of the various complications of modern medicine, died last year. This caused us to re-read and to remind the readers about the various ingredients of this iatrogenic phenomenon: polyphragmasia, the false diagnosis of "non-diseases", over-activity and over confidence of the medical establishment, non-adherence to logical work up of diagnostic algorithm, etc. Although medicine has progressed immensely, we often encounter the various maladies of modern medicine. In rehabilitation medicine practice, we also often face a few more aspects of this subject. PMID- 15116591 TI - [The annual scientific meeting of the Israel Society of Rheumatology in Ashkelon, December 18-19th, 2003]. PMID- 15116592 TI - Foster G. McGaw Prize. Phoebe Putney Memorial Hospital's "upstream investment" wins prestigious national award. PMID- 15116593 TI - Memorial Health University Medical Center named in Fortune's list of best companies to work for. PMID- 15116594 TI - OIG issues additional advisory opinions. PMID- 15116595 TI - Glenoid resurfacing in shoulder arthroplasty: indications and contraindications. AB - The indications for glenoid resurfacing are controversial. Advantages of glenoid resurfacing include decreased glenoid pain from metal-on-bone articulation, increased stability provided by the conforming glenoid component in the presence of asymmetric glenoid wear, and lateralization of the joint line providing for improved range of motion and strength. Proponents of hemiarthroplasty claim that function and pain relief are equivalent to glenoid resurfacing without the concomitant risk of glenoid loosening and loss of glenoid bone stock. In addition, hemiarthroplasty requires less surgical time and is less expensive than total shoulder arthroplasty. Total shoulder arthroplasty has been shown to be superior to hemiarthroplasty with regard to pain relief, range of motion, and function. Resurfacing of the glenoid has been indicated in older patients with primary osteoarthritis. In addition, patients with rheumatoid arthritis generally have better improvement in pain and function when treated with total shoulder arthroplasty than with hemiarthroplasty. Patients with severe rotator cuff disease, persistent instability, and lack of glenoid bone stock should be treated with hemiarthroplasty alone because glenoid resurfacing is associated with early loosening and failure. Patients with osteonecrosis isolated to the humeral head should be treated with hemiarthroplasty to preserve the native, congruent glenoid. PMID- 15116596 TI - Arthroscopic acromioplasty: history, rationale, and technique. AB - Subacromial impingement or supraspinatus syndrome has been known to exist in some form since the early 1900s. Several early studies have discussed subacromial impingement or supraspinatus syndrome. Arthroscopic acromioplasty has evolved as a useful surgical treatment. Pertinent physical examination and radiographic evaluation findings must be identified. In order for the surgery to be successful, it is important to understand the rationale for treatment of impingement syndrome and the technical aspects of arthroscopic acromioplasty. PMID- 15116597 TI - Conservative surgical treatment of wrist arthritis. AB - The diagnosis and treatment of intermediate stage wrist arthritis remains a challenge. In the earliest stages, the goal of treating prearthritic conditions, such as carpal instability caused by ligament injuries or carpal or distal radius fractures, is clear: anatomic reduction and healing of the fracture and correction of the instability to forestall degeneration of the wrist. Similarly, in established arthritis with severe pain and limitation of motion caused by advanced destruction of the articular surface, treatment usually involves some form of arthroplasty or arthrodesis. However, the patient with wrist pain and established but mild, moderate, or localized degenerative changes and good motion remains a treatment dilemma. PMID- 15116598 TI - Osteoarthritis of the wrist. AB - Osteoarthritis of the wrist is one of the most common conditions encountered by the orthopaedic surgeon and requires careful analysis and specific indications and rationale for appropriate treatment. Radiographic analysis and surgical approaches for reconstruction of the wrist consist of technical caveats that will improve outcome, including limited wrist fusions, proximal row carpectomy, and total wrist fusion. PMID- 15116599 TI - Surgical management of the arthritic wrist. AB - Wrist destruction by rheumatoid arthritis adversely affects hand function and increases the risk of finger deformities. Surgical management is indicated not only for salvage of end-stage wrist deformity but to alleviate pain and reduce progression of deformity at earlier stages. Available procedures include arthroscopic synovectomy, partial and complete arthrodesis, and implant arthroplasty. Procedure selection is highly dependent on disease characteristics and patient needs. PMID- 15116600 TI - Bearing surface options for total hip replacement in young patients. AB - Encouraging results and new implant developments have allowed total hip replacement to be performed in increasingly younger and more active patients. In young patients, however, outcomes are not comparable to those seen in older patients. The inflammatory reaction to polyethylene wear particles is one of the main causes of aseptic loosening and subsequent revision surgery and can limit the longevity of an arthroplasty in young and active patients. The wear resistance of polyethylene has been improved by cross-linking; however, greater degrees of cross-linking are associated with progressive decreases in other material properties, which theoretically increase the risk of component failure from very high or localized stresses. Ceramic femoral heads have been associated with lower in vivo polyethylene wear rates, which have been variable and up to 50% lower than with metallic heads. Metal-on-metal bearings have been reintroduced with improved materials, design, and manufacturing, although theoretical concerns remain regarding long-term exposure to metal particles and ions. Improved ceramic-on-ceramic bearings also are available and have the lowest wear rates. They offer a very small risk of in vivo fracture; however, they are the most expensive bearing option and wear rate is more sensitive to implant position. Long-term clinical studies are needed to show a reduction in revision surgery associated with the use of this current generation of bearings. The use of any of these bearings has specific benefits and risks that should be considered on a patient-by-patient basis. PMID- 15116601 TI - The diagnosis and treatment of nontraumatic osteonecrosis of the femoral head. AB - Osteonecrosis of the femoral head is a potentially debilitating disease that frequently affects young patients in the third through fifth decades of life. If untreated, this disease will result in total destruction of the hip joint; therefore, early diagnosis and intervention are essential to optimize outcome. Although the etiology of osteonecrosis of the femoral head is poorly understood, awareness of well-established risk factors and associated disorders can assist in early detection and possible prevention of hip joint destruction. All patients with symptomatic hip disease should undergo standard radiographic evaluation; however, MRI remains the most sensitive diagnostic modality. Depending on the stage of the disease, options for treatment range from minimally invasive procedures such as core decompression to total joint arthroplasty. PMID- 15116602 TI - Evaluation of the unstable total hip arthroplasty. AB - Despite advances in total hip arthroplasty, dislocation persists as a troublesome complication for orthopaedic surgeons to manage, second only to prosthetic loosening as a cause of revision. Although this complication has received considerable attention, evaluation and treatment of the unstable total hip arthroplasty remain poorly understood. Therefore, it is important to evaluate such factors as underlying patient comorbidities, the direction of dislocation, soft-tissue tension, surgical technique, implant design, and component position. For recurrent hip instability, a careful history to determine the mechanism of dislocation is necessary, and a review of preoperative imaging studies can help clarify whether gross component malpositioning is present. Examination under anesthesia and intraoperative inspection are also important. Taking the hip through a full range of motion while directly visualizing the anatomy can help diagnose component-to-component impingement, inadequate offset soft-tissue tension, extra-articular impingement, and other possible contributing factors. Even when a seemingly obvious cause of dislocation, such as component malposition, is diagnosed, surgical results have been somewhat disappointing. Therefore, surgical planning should include all possible revision options, and the temptation to find the quick fix should be resisted. Patients should be extensively counseled regarding realistic expectations both before primary hip arthroplasty and in the face of a revision surgery for recurrent dislocations because long-term results are less than optimal. PMID- 15116603 TI - Periprosthetic fractures of the acetabulum. AB - Periprosthetic fractures of the acetabulum after total hip arthroplasty are uncommon, but are increasing in number and severity. These fractures may occur intraoperatively, during the perioperative period, or many years after the total hip arthroplasty. Periprosthetic fractures of the acetabulum vary in severity and may involve stress fractures of the pubis or medial wall, significant bone loss secondary to osteolysis and subsequent loss of column integrity, or complete pelvic discontinuity. Treatment differs depending on the complexity of the fracture and the stability of the acetabular prosthesis. Surgical treatment for an unstable acetabulum should stabilize the bony columns of the acetabulum, provide bone grafting of defects, and should maintain adequate bone stock for replacement of a stable acetabular implant. Strict adherence to the principles of fracture surgery is required to achieve bony union of the acetabular columns and provide a stable environment for reimplantation of an acetabular component. PMID- 15116604 TI - Cable plates and onlay allografts in periprosthetic femoral fractures after hip replacement: laboratory and clinical observations. AB - Fractures of the femur after total hip replacement are an increasingly common and technically challenging problem. The results of nonsurgical treatment are poor. When the general condition of the patient allows, these injuries should be treated surgically. Several surgical treatments can be used to treat these fractures, and classification of the fracture assists the surgeon in the choice of procedure. Over the past decade, cable plate fixation systems and onlay strut allografts have become two of the most commonly used methods of fixation for fractures associated with hip prostheses. The ideal method of fixation is yet to be determined. However, laboratory studies have shown that dual fixation using either a lateral plate and anterior strut graft or two strut grafts produces the strongest construct. Cables rather than smooth wires should be used for fixation, and fixation strength increases with the number of cables used. The use of screws proximally produces a strong fixation but there are theoretical disadvantages to using screws around a femoral implant. Clinical data show high rates of fracture union using cable plate fixation, cortical onlay allograft fixation, and combined fixation methods for fractures that occur around well-fixed implants. Results have been less encouraging when these techniques have been used to fix fractures around prostheses that are either loose or malaligned; such fractures are better managed by revision of the femoral component to a long-stemmed device. Care should also be taken when there has been previous periosteal stripping of the femur because this may predispose to fracture nonunion. Periprosthetic fractures of the femur are a complex surgical problem and require specialized training in a range of surgical techniques. PMID- 15116605 TI - Periprosthetic fractures of the femur after total hip arthroplasty. AB - The incidence of periprosthetic femur fracture has increased recently, and these fractures have become of a great concern to the reconstructive orthopaedic surgeon. Intraoperative fractures are usually stable. To prevent intraoperative fracture, careful preoperative planning and gentle surgical techniques are essential. In managing unstable intraoperative and late postoperative periprosthetic fractures, the surgeon should know the exact pattern of fracture, prosthesis stability, and bone quality. Loose prostheses should be revised and displaced fractures should be reduced and adequately fixed. PMID- 15116606 TI - Extended trochanteric osteotomy: planning, surgical technique, and pitfalls. AB - The extended trochanteric osteotomy is appropriate for a number of surgical indications. It facilitates removal of well-fixed cement mantles with a loose or well-fixed stem and of extensively porous-coated or tapered cementless stems. This exposure is particularly valuable in the presence of varus remodeling of the proximal femur, permitting correction of proximal femoral deformity and reducing the risk of fracture of the greater trochanter. It is also indicated in the removal of a well-fixed cemented stem that is complicated by infection, where it is vital to extract all foreign material for successful eradication of the infection. In addition, when the osteotomy is required for femoral exposure, it enhances acetabular exposure to allow even the most complex reconstruction. Also, as the soft-tissue attachments to the bone fragment are preserved in this approach, abductor muscle tension can be adjusted. PMID- 15116607 TI - Cementing constrained acetabular liners in revision hip replacement: clinical and laboratory observations. AB - During revision hip arthroplasty, removal of a well-fixed, ingrown metal acetabular component may not be possible. Therefore, a new polyethylene liner can be cemented into the existing shell via the cement locking mechanism. This technique is well recognized, and the cement locking mechanism has proved to be sufficiently strong and durable for clinical use. A constrained polyethylene liner is designed to reduce the risk of hip dislocation by capturing the femoral head. However, there are increased shear forces created at the liner interface as the dislocation is resisted. If a constrained liner is cemented into an ingrown acetabular component, then there is the theoretical risk that these increased shear forces will damage the cement locking mechanism, thus leading to failure of the construct. There are a few clinical series in which a constrained liner has been used with the cement locking mechanism. Overall, the failure rate of the cement locking mechanism is no greater if a constrained liner instead of a standard liner is used. The cement locking mechanism can be strengthened by roughening the backside of a smooth polyethylene liner to improve the cement polyethylene interface, or by using an all-polyethylene acetabular component that is designed to be used with cement. Whether a smooth metal shell needs to be roughened as well is a matter of debate. PMID- 15116608 TI - The mini-incision approach to total hip arthroplasty. AB - Total hip arthroplasty (THA) usually involves an incision of approximately 20 to 25 cm long. Surgeons recently have begun to question the need for such a long incision and are developing less invasive approaches to THA. A mini-incision approach to THA promotes shorter hospital stays, faster rehabilitation, and an increase in patient satisfaction without a detrimental effect on outcome. The surgical technique is based on the classic Moore approach to the hip. The incisions measure 6 to 10 cm in length, and the surgery involves significantly less deep soft-tissue disruption. All the basic tenets of THA are respected and visualization is maintained throughout the procedure for proper placement of the components. Mini-incision THA offers a safe and effective alternative to THA performed through a larger incision. Obese patients typically are not good candidates. Patients with severe hip dysplasia or those undergoing revision surgery require a wider exposure to perform the arthroplasty to ensure the best possible result. In properly selected patients, however, THA can be performed through an abridged mini-incision without compromising the results or exposing the patient to additional risk. PMID- 15116609 TI - The technique of minimally invasive total hip arthroplasty using the two-incision approach. AB - The two-incision technique for total hip arthroplasty is a minimally invasive procedure that avoids the transection of any muscle or tendon. This technique has the potential to reduce surgical trauma, pain, and morbidity and to speed patient recovery. Fluoroscopy-assisted, two-incision total hip arthroplasty uses several new instruments that have been developed to facilitate exposure of the surgical site and component placement. Standard implants with well-established designs are used to maintain the present expectation for implant durability. PMID- 15116610 TI - Less invasive total hip arthroplasty using navigational tools. AB - Although traditional total hip arthroplasty offers good visualization of bony landmarks and allows for the accurate orientation and fixation of implants, these benefits are achieved at the expense of extensive soft-tissue dissection and can result in postoperative complications and a delayed return to full function. To address these disadvantages, navigational tools were coupled with a mini-incision technique that allowed accurate bone preparation and orientation of the implant components without direct visualization of the bony landmarks. Additionally, image-guided systems provide three-dimensional information before and during surgery, making it possible to know, in real time, the orientation of implants and to visualize the full bony anatomy. This "computer-enhanced vision" allows surgeons to perform less invasive and eventually minimally invasive total hip arthroplasty with improved accuracy. PMID- 15116611 TI - Indications for patellar resurfacing in total knee arthroplasty. AB - The management of the patella in total knee arthroplasty (TKA) traditionally has been one of three options: always resurface, never resurface, or selectively resurface the patella. Historically, implant design and surgical technique did not completely address the patellofemoral articulation. Increased understanding of patellofemoral anatomy, biomechanics, implant design, and surgical technique have led to an improvement in the previously reported high rate of patellofemoral complications associated with TKA. Traditional indications for patellar resurfacing, including age, weight, gender, patellar anatomy, quality of articular cartilage, radiographic findings, and the presence of rheumatoid arthritis deformity and preoperative anterior knee pain continue to be debated. Anterior knee pain before and after TKA must not always be presumed to be secondary to a patellofemoral resurfacing/nonresurfacing etiology, and other factors may play a role in the dynamic development of anterior knee pain after TKA. The decision to resurface the patella in TKA remains controversial, and the results of longer-term randomized controlled trials will improve understanding of this complex issue in the future. PMID- 15116612 TI - Implant design and techniques for patellar resurfacing in total knee arthroplasty. AB - There are two basic techniques for resurfacing of the patella in total knee arthroplasty. The inset patellar component recesses a circular dome with a single fixation peg into a reamed patella while the onlay component is placed onto the cut surface of patella. Onlay patellar components have a wide variety of shapes, designs, and methods of fixation. There are differences in technique and possible complications with the inset and onlay patellar components. However, neither method has proved to be superior. Fixation of an onlay patellar component is usually with cement, but successful cementless porous-coated designs exist. Fixation of an all-polyethylene patellar component occurs by cementing a component with either a single central peg or three smaller peripheral pegs; although one method is not preferred over the other, there have been reports of breakage with three pegs. The design and fabrication of these components are important to prevent peg breakage with repeated shear stress. There are few clinical data implicating the different shapes or methods of patella component fixation with the complications of anterior knee pain, wear, patella fracture, and component breakage. Attention to detail during surgery is likely the most critical factor to the success of patellar resurfacing in total knee arthroplasty. PMID- 15116613 TI - Extensor mechanism complications in total knee arthroplasty. AB - Recent improvements in total knee arthroplasty (TKA) surgical technique, instrumentation, and prosthetic design have decreased the incidence of extensor mechanism complications following TKA. The majority of these complications continue to be secondary to errors in surgical technique. Extensor mechanism complications following TKA include soft-tissue impingement, instability, patellar fractures, patellar component failures, and extensor mechanism rupture. Both nonsurgical and surgical treatment may be used in the management of these complications. However, the majority of these complications are avoidable with attention to proper surgical technique and selection of appropriate patellar components in TKA. PMID- 15116614 TI - Revision of failed patellar components. AB - In the majority of revision knee replacements, the existing patellar component is well fixed and it is possible to retain the existing patellar component when specific criteria are met. If the patellar component is loose, malpositioned, or damaged, it should be revised. The decision regarding whether to implant another prosthesis or to perform an alternative procedure is dependent on an assessment of remaining patellar bone stock. Patellae with severe bone deficiency, which preclude adequate fixation of another patellar implant, occur in approximately 10% of revision knee replacements. Specific techniques include patellar resection arthroplasty, the gull-wing osteotomy, structural bone grafting of the patella, morcellized bone grafting of the patella, or use of a porous metal augmentation baseplate. Whether the patellar revision being performed is an isolated procedure or is part of revision arthroplasty, optimal positioning of the femoral and tibial component is important for proper patellar kinematics. PMID- 15116615 TI - Managing instability in total knee arthroplasty with constrained and linked implants. AB - Instability is a relatively common cause of failure of total knee arthroplasty. In the management of the unstable total knee, the need for constraint must be anticipated. Careful preoperative planning including history and physical examination of the ligamentous support of the knee is mandatory. A thorough study of current and previous radiographs is important to determine wear, component migration, and bone loss. As a general rule, it is recommended that the minimum amount of constraint necessary to achieve stability should be used. Most of the time, stability can be achieved using a posterior-stabilized or a nonlinked implant, such as a varus-valgus constrained or constrained-condylar knee implant. Occasionally, a hinge prosthesis may be indicated, particularly in a patient with an absent and nonreconstructable medial collateral ligament. Increasing constraint is not without its problems, however, as forces across the knee may be transmitted to the stem-bone interface, resulting in radiographic loosening of stemmed components. Fortunately, these observations of radiographic loosening around stems of revision components have not so far correlated with reports of clinical failure. PMID- 15116616 TI - Periprosthetic fractures of the tibia and patella in total knee arthroplasty. AB - Periprosthetic fractures of the tibia and patella associated with total knee arthroplasty are less common than periprosthetic fractures of the distal femur and therefore have been less frequently reported in the literature. Because more total knee replacements are now being performed and implant and patient survival has improved, periprosthetic fractures of the tibia and patella may be recognized more frequently in association with both primary and revision total knee replacement. Several risk factors associated with both types of fractures have been identified, and addressing patient factors, surgical techniques, and implant designs that may contribute to increased risk is an important aspect of management. PMID- 15116617 TI - Treatment of the unstable total knee arthroplasty. AB - Instability is one of the leading causes of clinical failure after total knee arthroplasty. Instability can be categorized according to three basic patterns: AP or flexion space instability, varus/valgus or extension space instability, and global instability. Surgical options for treating instability include polyethylene exchange of a modular component, revision to a more constrained component, or revision to a hinged component. The results achieved with these surgical options vary for each type of instability. In general, the use of more constrained components to manage unstable knees is favored. PMID- 15116618 TI - Epidemiology of osteolysis: backside implant wear. AB - Osteolysis is an emerging problem in patients who undergo total hip arthroplasty and total knee arthroplasty (TKA). Before the introduction of modular components, osteolysis was not as prevalent in patients who underwent TKA. Although polyethylene wear and the generation of small-particle debris were linked to the presence of osteolysis in patients who have undergone total hip arthroplasty, the fatigue-type wear and larger wear particles often seen with TKA were not thought to invoke the cellular response that causes osteolysis. Despite the lack of an identifiable cause for the osteolysis, the number of knees that developed lesions continued to increase. Therefore, modular components were examined to determine whether this innovation in TKA was actually contributing to the manifestation of osteolysis. PMID- 15116619 TI - Sources of osteolysis around total knee arthroplasty: wear of the bearing surface. AB - Once mostly a problem in total hip arthroplasty, osteolysis has now emerged as a significant problem around total knee arthroplasty (TKA). The development of osteolysis is often related to wear of the polyethylene bearing surface with production of biologically active particulate debris. There are many structural and design factors related to the polyethylene bearing surface that have been shown to affect the extent of wear that occurs over time. For instance, the processing, manufacturing, and sterilization methods of polyethylene all are critical factors in determining its mechanical and wear-resistant properties. Cross-linking of polyethylene bearing surfaces in TKA, however, may not demonstrate the same degree of improved resistance to wear as seen in total hip arthroplasty. Optimization of polyethylene bearing surface thickness, with a goal to insert at least 8 mm of polyethylene, has been shown to minimize contact stresses and subsequent fatigue failure of total knee implants. Selecting a well designed component with minimal counter surface roughness is also important in minimizing the generation of polyethylene wear debris and subsequent osteolysis. Finally, proper surgical technique, including restoration of mechanical axis and rotational alignment, is critical to minimizing wear and maximizing the longevity of TKA. PMID- 15116620 TI - Recognizing and identifying osteolysis around total knee arthroplasty. AB - Osteolysis around total knee arthroplasty can be difficult to identify because it typically occurs in cancellous bone of the distal femur and proximal tibia, which has a low radiodensity, and because it is often obscured on radiographs by the metallic prosthesis. Careful evaluation is warranted in circumstances where osteolysis is likely: radiographically evident polyethylene wear, high-activity young patients with a prosthesis in place for a long time period, or implant designs associated with osteolysis. Osteolysis of the distal femur is best seen on lateral radiographs and often involves the posterior condyles; on anteroposterior radiographs it may be seen as radiolucency extending proximally into either condyle. Osteolysis of the tibia often occurs along access tracks such as screws or around the periphery of well-fixed implants. Three-dimensional imaging modalities with new metal suppression technology hold the promise of improving accuracy of osteolysis detection in the near future. PMID- 15116621 TI - Different surgical options for monocompartmental osteoarthritis of the knee: high tibial osteotomy versus unicompartmental knee arthroplasty versus total knee arthroplasty: indications, techniques, results, and controversies. AB - The decision of what procedure to perform for the treatment of monocompartmental osteoarthritis of the knee when nonsurgical treatment methods fail remains controversial. Recent advances using osteotomy, unicompartmental knee replacement, and total knee replacement have been reported. For example, there are new concepts for performing high tibial osteotomies rather than the traditional Coventry method. Many techniques now involve osteotomies below the tibial tubercle. Unicompartmental knee replacement can be done using a standard approach, but less invasive approaches exist, along with minimally invasive approaches for total knee replacement, rather than the standard large incision, that promote decreased soft-tissue destruction. PMID- 15116622 TI - Keratotic disorders of the plantar skin. AB - Keratotic lesions on the plantar aspect of the foot develop beneath an osseous prominence and can result in substantial disability. This occurs because, during normal gait, the metatarsal head area is subjected to more prolonged stress than any other area on the plantar aspect of the foot. In the treatment of this disorder, it is imperative to establish the etiology, among many possibilities, and then address the specific pathology accordingly. PMID- 15116623 TI - The bunionette deformity. AB - The bunionette deformity results in pain about the lateral and/or the plantar aspect of the fifth metatarsal head. It is important to carefully assess the deformity anatomically in order to select the proper surgical procedure to correct this painful affliction. Knowledge of pathoanatomy and certain surgical procedures can be used to correct the deformity. PMID- 15116624 TI - Evaluation and treatment of chronic ankle pain. AB - The evaluation and treatment of chronic ankle pain presents a challenge to the orthopaedic surgeon. A detailed history helps to determine causative factors resulting from earlier trauma or surgery. A careful physical examination and radiographic studies also are helpful in making an accurate diagnosis, which is the basis for choosing a specific and effective treatment regimen. PMID- 15116625 TI - Lumbar arthrodesis for degenerative conditions. AB - There is significant disagreement among spine surgeons regarding the optimal technique of arthrodesis for treatment of degenerative disorders of the lumbar spine. Degenerative conditions of the lumbar spine include degenerative disk "disease," post-decompression degeneration, degenerative spondylolisthesis, junctional degeneration, spondylolis, and low-grade lytic spondylolisthesis. Although it is impossible to develop strict evidence-based criteria for the selection of one surgical approach over another, some generalizations are possible based on empiric process, anecdotal experience, and published surgical series. Patient selection, cessation of nicotine use, and use of autologous bone graft are factors that influence clinical outcome after lumbar arthrodesis. PMID- 15116626 TI - Cervical spine and spinal cord injuries: recognition and treatment. AB - Cervical spine injuries are frequently encountered in any practice treating patients in the emergency department or outpatient trauma victims. When upper or lower cervical spine injuries are suspected, immediate immobilization, physical and neurologic examination, and radiographic evaluation are imperative. For spinal cord injuries, knowledge of microscopic and cellular pathology helps to determine appropriate management. PMID- 15116627 TI - Diagnosis and management of thoracolumbar spine fractures. AB - The lack of robust clinical studies has contributed to controversy regarding optimal treatment for patients with injuries to the thoracolumbar spine. The transitional anatomy of the thoracolumbar spine makes it vulnerable to injury resulting from high-energy motor vehicle collisions and falls; osteoporosis is an underlying factor in most of the compression fractures identified in elderly patients. The formulation of a treatment plan for patients with injuries to the thoracolumbar spine depends on the presence and extent of neurologic injury and deformity and an estimate concerning spinal stability. Both nonsurgical and surgical treatment options are available to achieve the goals of preservation of neurologic function and restoration of spinal stability. PMID- 15116628 TI - Diagnosis and management of sacral spine fractures. AB - The sacrum is the mechanical hub of the axial skeleton, serving as the base for the spinal column and keystone for the pelvic ring. Both surgical and nonsurgical options are available to treat sacral spine fractures; however, because these fractures are relatively rare and heterogeneous in nature and because there is little evidence-based literature, choosing the optimal treatment is challenging. The timing of intervention and the choice of surgical technique need to be determined on an individual basis, with the goal of producing the best outcome for the patient. PMID- 15116629 TI - Current concepts in the treatment of distal radial fractures. AB - Surgical indications for the treatment of distal radial fractures are evolving. It is important to identify the various articular fragments and their significance to facilitate optimal surgical treatment of these fragments from the standpoint of both internal and external fixation. New techniques in the visualization and stabilization of the articular surface and the treatment of defects in the metaphysis, including the use of cement to buttress the articular surface, have been brought to the forefront. A treatment algorithm for associated injuries to the distal radioulnar joint is also helpful. PMID- 15116631 TI - Prevention of hip fractures: medical and nonmedical management. AB - Hip fractures can have a significant detrimental effect on morbidity and mortality. Medical and nonmedical management approaches both may be used to help decrease the risk of hip fracture. Medical management includes the use of antiresorptive agents such as the bisphosphonates, calcium and vitamin D, selective estrogen receptor modulators, and anabolic agents such as parathyroid hormone, which strengthen bone. Nonmedical management includes fall prevention programs and hip protectors. Physicians caring for patients at risk for hip fracture should be cognizant of these management approaches to most effectively minimize fracture risk. PMID- 15116630 TI - Nonprosthetic management of proximal humeral fractures. AB - Many proximal humeral fractures can be treated without the need for hemiarthroplasty. Treatment choice is affected by fracture location and pattern, as well as by patient factors including age, activity level, quality of bone, and ability to comply with a regimen of therapy. Successful diagnosis and treatment of proximal humeral fractures is dependent on good-quality radiographs, but in some cases, intraoperative assessment of the fracture pattern is required for a complete and accurate diagnosis of the fracture pattern and severity. A discussion of nonsurgical and surgical treatment options and techniques needed to achieve anatomic reduction and stable fixation is important. PMID- 15116633 TI - Osteoporotic pertrochanteric hip fractures: management and current controversies. AB - The treatment of intertrochanteric hip fractures has evolved over the past 80 years because of a better understanding of fracture anatomy, application of biomechanical principles, and novel technologic advances. Surgical treatment of intertrochanteric hip fractures is the current standard of care, with short-term goals of fracture stabilization and early patient mobilization and the long-term objective of restoring patients to their previous level of independence and function. Treatment for stable intertrochanteric hip fractures includes use of percutaneous intramedullary devices and open reduction and internal fixation using a sliding hip screw. To date, none of these devices has shown any clear clinical advantage over the dynamic hip screw. Intramedullary fixation has multiple theoretical advantages for the treatment of unstable fracture patterns; however, it remains unclear if ultimate functional outcome warrants the added expense of such treatment. For patients with a reverse obliquity fracture pattern, the advantage of the intramedullary construct has been shown. PMID- 15116632 TI - Osteoporotic femoral neck fractures: management and current controversies. AB - Osteoporosis is a pervasive disease among the growing elderly population. Femoral neck fractures are often a direct result of osteoporosis and are challenging to treat. Surgical interventions seek to return the patient to preinjury function as quickly as possible, but many obstacles exist. Disruption of the blood supply occurs regardless of the fracture pattern, and in the active elderly population, reduction and fixation should be done as soon as possible to minimize healing problems. Closed reduction with percutaneous cannulated screw instrumentation is currently the fixation method of choice, but even with meticulous technique, moderate complication rates persist. Newer devices and biologic bone augmentation cement show promise in decreasing postoperative fracture collapse. Patients in whom a stable reduction cannot be achieved or who have a limited life expectancy should undergo arthroplasty. Unipolar and bipolar arthroplasty have both been effective in restoring function and have been the standard of care in these patients. Recent evidence suggests that active elderly patients who have acetabular disease or severely displaced fractures may benefit most from primary total hip arthroplasty. PMID- 15116634 TI - Management of troublesome stress fractures. AB - Stress fractures can be challenging to treat. It is important to educate the clinician in the pathophysiology, etiology, evaluation, and treatment of stress fractures with special emphasis on troublesome and upper extremity stress fractures, along with the pathophysiology of fatigue failure of bone and the etiology of clinically significant stress fractures. A classification system has been outlined to facilitate management of troublesome stress fractures. PMID- 15116635 TI - Minimally invasive percutaneous plate osteosynthesis of fractures of the distal tibia. AB - Fractures of the distal tibia are notoriously difficult to treat, and traditional methods of fixation are often fraught with soft-tissue complications. With recent emphasis on meticulous handling and preservation of the soft-tissue envelope, minimally invasive percutaneous plate osteosynthesis has become a safe and reliable method of treating these fractures. This technique involves conventional open reduction and internal fixation of the fibula and spanning external fixation of the tibia until the soft-tissue swelling subsides. Subsequently, limited open reduction and internal fixation of displaced articular fragments is performed through small incisions based on CT evaluation. This is followed by minimally invasive percutaneous plate osteosynthesis of the tibia, in which the plafond is attached to the tibial shaft using a variety of commercially available plates. PMID- 15116636 TI - Kyphosis of the thoracic and thoracolumbar spine in the pediatric patient: normal sagittal parameters and scope of the problem. AB - As measured by the Cobb angle, normal sagittal kyphosis is 20 degrees to 40 degrees, which encompasses most of the angulated consecutive vertebrae in the thoracic region of the spine. With pathologic kyphosis, however, the segmental analysis of different regions of the thoracic spine plays an important role. Methods of determining sagittal measurements as well as the causes of kyphosis of the thoracic and thoracolumbar spine, including Scheuermann's disease, spinal cord injury, laminectomy, neurofibromatosis, genetic origins, Marfan syndrome, and tuberculosis, are also important in determining treatment. PMID- 15116637 TI - Nonsurgical treatment of kyphosis. AB - An increase in thoracic kyphosis in children and adolescents is usually the result of postural kyphosis or Scheuermann's kyphosis. Although no structural deformity of the spine is observed in postural kyphosis, wedging of vertebral bodies and disk space narrowing are noted radiographically in patients with Scheuermann's kyphosis. Effective interventions for adolescents with postural kyphosis include exercises to relieve lower extremity contractures and strengthen abdominal musculature coupled with practiced normal posture in stance and in sitting. Skeletally immature patients with Scheuermann's kyphosis benefit from a similar exercise program but also require the use of a spinal orthosis. Bracing of the spine in patients with Scheuermann's kyphosis results in permanent correction of vertebral deformity, unlike bracing in patients with idiopathic scoliosis. The evaluation of children and adolescents with increased thoracic kyphosis is an important aspect of the decision process used to determine appropriate interventions. PMID- 15116638 TI - Kyphosis of the thoracic and thoracolumbar spine in the pediatric patient: surgical treatment. AB - Kyphosis of the thoracic or thoracolumbar spine is a common deformity in pediatric and adolescent populations. When it progresses to the point at which nonsurgical treatment is no longer an option, surgery is indicated. Surgical options available for the treatment of different types of pediatric kyphosis of the thoracic and thoracolumbar spine include posterior instrumentation and fusion, posterior instrumentation and fusion combined with anterior fusion, and anterior instrumentation and fusion. PMID- 15116639 TI - Kyphosis of the thoracic and thoracolumbar spine in the pediatric patient: prevention and treatment of surgical complications. AB - The successful outcome of surgical treatment of pediatric kyphosis depends on careful preoperative identification of the etiology of the problem as well as identification of any associated medical conditions or syndromes that may render surgical reconstruction more challenging. Many perioperative surgical factors can lead to an unsuccessful surgical outcome, including inadequate preoperative patient or kyphosis assessment; inappropriate selection of proximal and/or distal instrumentation and fusion levels; inadequate spinal fixation applied at the ends of the posterior construct where tension forces are greatest; inadequate performance of a meticulous posterior spinal fusion; absence and/or an inadequate performance of an anterior spinal fusion when required either before or after the posterior procedure; overcorrection of the kyphotic deformity based on the ability of the spine above and below to compensate for the correction; a higher risk of neurologic complications with correction of kyphotic deformities; and inadequate postoperative support with an orthosis. Revision surgery for failed pediatric kyphosis surgeries requires careful reexamination of all these factors to correct any shortcomings. In addition, adjunctive procedures such as spinal osteotomies, perioperative traction, and/or anterior fusion techniques may be required to optimize spinal alignment, balance, and ultimate successful fusion. When following these guidelines, pediatric spinal kyphosis disorders can be successfully treated and complications avoided. PMID- 15116640 TI - Optimization of walking ability of children with cerebral palsy. AB - A new paradigm based on an appreciation of the biomechanics of normal and pathologic gait and a better understanding of muscle-tendon unit anatomy and physiology has emerged for orthopaedic clinical decision making to optimize the ambulatory abilities of children with cerebral palsy. This quantitative, biomechanically based approach has been accepted as a research and teaching tool and as an instrument of outcome assessment; however, controversy remains concerning the expense of using this approach and about its accuracy and repeatability. This paradigm is used within a diagnostic matrix consisting of five data sources. Members of the clinical and technical teams from the motion analysis laboratory interpret data from the clinical history, physical examination, diagnostic imaging, quantitative gait analysis, and examination under anesthesia. The certainty of intervention selection is proportional to the consistency of the data within the diagnostic matrix. When inconsistencies in the data exist, input from both the clinical and technical teams is needed to resolve discrepancies. Working within the framework of the diagnostic matrix, it is possible to identify the indications used in the selection and recommendation of musculoskeletal surgical interventions to optimize gait in children with cerebral palsy. It is important to examine indications and controversies for surgical intervention related to iliopsoas recession, femoral rotational osteotomy, medial hamstring lengthening, rectus femoris transfer, and gastrocnemius recession. PMID- 15116641 TI - Developmental hip dysplasia and dislocation: Part I. AB - A thorough knowledge of the normal growth and development of the hip, the causes of abnormal development, and the structural and functional changes that result from developmental hip dysplasia and dislocation provide needed information for treating these conditions. Ultrasonography, newborn screening, and radiographic evaluation are important diagnostic tools. PMID- 15116642 TI - Developmental hip dysplasia and dislocation: Part II. AB - Both nonsurgical and surgical options are available for the treatment of developmental hip dysplasia and dislocation. The advantages, pitfalls, and techniques for using the Pavlik harness should be thoroughly examined before treatment. Other closed and open treatments are aimed at concentric reduction and prevention of residual subluxation and dysplasia. Early diagnosis and treatment lead to the best long-term results for these conditions. PMID- 15116643 TI - Arthroscopic treatment of anterior glenohumeral instability: indications and techniques. AB - The arthroscopic treatment of anterior glenohumeral instability is becoming increasingly accepted as a viable treatment option because reported success rates parallel those of open stabilization techniques. This improved success rate is largely the result of advances in surgical techniques and technology. An improved understanding of the pathoanatomy associated with shoulder instability and continuing education initiatives have also been instrumental in expanding the indications for arthroscopic stabilization of the unstable shoulder. Important considerations during arthroscopy include identifying all pathology, mobilizing soft tissue, enhancing the local biology to promote soft-tissue healing to bone or to itself, securing anatomic fixation, and respecting the healing period during postoperative rehabilitation efforts. Principal contraindications include significant bone deficits and the inability to repair capsular avulsions or rupture. Adherence to these basic principles should lead to excellent results with arthroscopic stabilization of the unstable shoulder. PMID- 15116644 TI - Arthroscopic versus open treatment of anterior shoulder instability. AB - Open repair is the accepted mainstay of treatment for anterior shoulder instability, and consistently good clinical results have been reported in the literature. With the development of newer arthroscopic imbrication techniques, however, arthroscopy may eventually provide equivalent long-term efficacy. PMID- 15116645 TI - Multidirectional instability: surgical decision making. AB - Although previous authors have described multidirectional instability of the shoulder, it was not until 1980 that Neer and associates solidified the current understanding and treatment of inferior and multidirectional instability. They emphasized the importance of differentiating this condition from the more common unidirectional instabilities and introduced the concept of an inferior capsular shift to globally tension the capsule anteriorly, inferiorly, and posteriorly, while thickening and reinforcing it on the side of greatest instability (i.e., anterior or posterior). Since the time of this initial description, the diagnosis and treatment of multidirectional instability has been fraught with difficulty and confusion. More recently, the advent of shoulder arthroscopy has blazed the trail for minimally invasive techniques to correct multidirectional instability, including arthroscopic thermal capsulorrhaphy and suture plication. However, despite strong opinions regarding these surgical techniques, when comparing open approaches to arthroscopic techniques for the treatment of multidirectional instability, long-term outcomes of arthroscopic techniques have yet to replicate those of open surgery. PMID- 15116646 TI - The diagnosis and treatment of labral and chondral injuries. AB - Hip arthroscopy is an excellent way to evaluate, diagnose, and treat acetabular labral and chondral lesions. It is minimally invasive and can be done on an outpatient basis. Candidates for hip arthroscopy will have functionally limiting symptoms and reproducible physical findings. Patients will often have mechanical symptoms such as clicking, catching, locking, or giving way. Symptoms may be preceded by a traumatic event such as a fall or twisting injury or may have an insidious onset. Radiographic studies are not sensitive enough to diagnose intra articular hip pathology; however, contrast agents used in conjunction with CT and MRI may aid in the diagnosis. Therefore, a high level of clinical suspicion and prudent clinical judgment are warranted. A thorough knowledge of positioning and anatomic relationships to portal placement is necessary to prevent potential neurovascular complications from occurring during hip arthroscopy. Labral tears are the most common cause of mechanical hip symptoms. Tears occur anteriorly, and associated chondral lesions are common. The severity of the chondral lesion is highly correlated with the surgical outcome. The most frequently observed chondral lesion is the watershed lesion, which consists of a labral tear with separation of the labrum from the articular surface at the labral-cartilage junction. The difficulty in identifying these lesions as well as their effect on outcome provides a convincing rationale for arthroscopic hip surgery. Arthroscopic treatment of these tears involves judicious debridement back to a stable base while carefully preserving the capsular labral tissue. Eliminating the source of mechanical symptoms secondary to labral pathology should alleviate the patient's discomfort. Chondral defects are drilled or treated with a microfracture technique to enhance fibrocartilage formation. Patient outcomes are directly dependent on the stage or extent of the labral and chondral lesion. PMID- 15116647 TI - Medial collateral ligament tears in the throwing athlete. AB - Medial collateral ligament (MCL) injuries of the elbow in throwing athletes are part of a spectrum of valgus extension overload injuries. Clinicians should consider reconstruction of the injured MCL in those patients who are unable to return to sports activities or work after an interval of rest and rehabilitation. Surgical treatment of this disorder has continued to evolve from the original technique developed by Jobe and associates. The authors' surgical technique involves reconstruction of the MCL using a single ulnar tunnel and single humeral tunnel that is performed through a muscle-splitting approach that usually obviates the need for ulnar nerve transposition. Knowledge of the biology and biomechanics of MCL function is important in the diagnosis and treatment of MCL injuries of the elbow in the throwing athlete. PMID- 15116648 TI - Tennis elbow tendinosis (epicondylitis). AB - Tennis elbow tendinosis (epicondylitis) is most commonly caused by tendon overuse and failed tendon healing. The pathoanatomy of overuse tendinopathy is noninflammatory "angiofibroblastic tendinosis." The specific areas of elbow abnormality include the extensor carpi radialis brevis-extensor digitorum communis complex laterally, the pronator teres, flexor carpi radialis medially, and triceps posteriorly. The primary goal of nonsurgical treatment is to revitalize the unhealthy tissue that produces pain. Successful nonsurgical treatment comprises rehabilitative resistance exercise and progression of the exercise program. If rehabilitation fails, surgical treatment can be quite successful. PMID- 15116649 TI - Osteochondritis dissecans of the elbow. AB - Osteochondritis dissecans of the elbow is a localized condition of the articular surface that is commonly seen in the young athlete. This disorder refers primarily to lesions of the capitellum and can be difficult to treat. Although trauma and ischemia play significant roles, the exact etiology remains unknown. The natural history is poorly understood and long-term sequelae include degenerative arthritis. The integrity of the articular surface and the stability of the lesion can be carefully evaluated with MRI and arthroscopy. Management is based mainly upon these two factors, yet no good universal outcomes exist among the varied treatment options. Stable lesions identified early appear to have the best prognosis with conservative management. Indications for surgery include persistent or worsening symptoms despite prolonged conservative care, loose bodies, or evidence of instability. Whether to excise and debride or to fix an unstable fragment is a highly controversial topic. The clinician should recognize osteochondritis dissecans of the elbow as a potentially disabling condition where the prognosis for return to sport is guarded. PMID- 15116650 TI - Posterolateral rotatory instability of the elbow. AB - Posterolateral rotatory instability (PLRI) of the elbow is recurrent elbow instability caused by injury to the radial ulnohumeral ligament or lateral ulnar collateral ligament. Deficiencies of the radial ulnohumeral ligament and laxity of the lateral capsule allow the proximal radioulnar joint to rotate and the radial head to subluxate posteriorly when stressed. Whether isolated or in conjunction with other injuries, PLRI represents the initial stage in the pathology of the unstable elbow. The diagnosis of PLRI can be difficult because the provocative tests are challenging to perform. Diagnostic arthroscopy is an excellent tool to demonstrate this instability. Treatment options include bracing for acute injuries, and primary repair or reconstruction for chronic injuries. Although arthroscopic plication and repair have been as effective as open techniques, the clinician should be prepared for open reconstruction if needed. PMID- 15116651 TI - Hip arthroscopy: when it is and when it is not indicated. AB - Problems that occur in the hip joint, such as a loose body or labral tear, can be treated arthroscopically or with conventional arthrotomy. Hip arthroscopy can facilitate both comprehensive access to and treatment of an evolving series of conditions that affect the hip joint. Candidates for hip arthroscopy should have reproducible symptoms and physical findings that limit function, and a history of mechanical symptoms. Following total hip replacement, when unexplained symptoms persist despite appropriate conservative treatment combined with a negative workup, arthroscopy can be valuable. Intra-articular third bodies often can be successfully removed arthroscopically. Arthroscopy can be valuable after trauma for evacuation of hematomas and removal of chondral loose bodies, and repair of labral injuries. Contraindications to arthroscopy include advanced osteoarthritis, osteonecrosis with femoral head collapse, an ankylosed joint, significant protrusio or grade III or IV heterotopic bone. PMID- 15116652 TI - Soft-tissue lumps and bumps. AB - Soft-tissue masses of the extremities and torso are a common problem encountered by the orthopaedic surgeon. Although these soft-tissue masses are often benign, the orthopaedic surgeon must be able to recognize the features key to differentiating benign and malignant masses. An understanding of the epidemiology and clinical presentation of soft-tissue masses is needed in order to outline a practical approach to evaluation and surgical management. PMID- 15116653 TI - The biopsy. AB - The biopsy of a musculoskeletal lesion is an important event, the outcome of which guides patient management and helps determine patient prognosis. The principles of biopsy include complete radiologic staging before the biopsy, thorough prebiopsy planning including consultation with the pathologist and radiologist, determining the most appropriate method of biopsy (fine needle, core needle, open surgical biopsy), placing the biopsy tract appropriately, and making sure the biopsy tract can be removed at the time of resection, avoiding contamination of uninvolved structures, avoiding transverse incisions, preventing pathologic fracture, handling biopsy tissue appropriately, and considering referral before biopsy. The common errors of biopsy include sampling errors, postbiopsy hematomas, the use of transverse incisions, tumor implantation, and the treatment of an unsuspected sarcoma with prophylactic fixation. Thoughtful prebiopsy planning and careful completion of the biopsy can result in an expedient and accurate diagnosis. If the treating physician lacks significant expertise in performing biopsy and management of patients with musculoskeletal lesions, then referral to a musculoskeletal oncologist before biopsy should be considered. PMID- 15116654 TI - Enchondroma and chondrosarcoma of bone: clinical, radiologic, and histologic differentiation. AB - Intraosseous cartilage tumors are commonly encountered by orthopaedic surgeons during evaluation of a patient for undetermined pain; however, the possibility of missing a malignancy during examination is cause for concern. Surgery for enchondromas is not generally necessary but when clinical and radiographic features suggest a potential chondrosarcoma, prompt referral is necessary. Chondrosarcoma almost always occurs in adults. Patients usually present with pain and have characteristic findings on staging studies. Lucencies developing within the calcification, periosteal reactions, and a soft-tissue mass all are characteristic features of chondrosarcoma. Biopsies are not typically indicated because of the histologic similarity of benign and malignant cartilage lesions. PMID- 15116655 TI - What to do with deep lipomatous tumors. AB - Although most deep soft-tissue sarcomas are fairly easily recognized as heterogenous masses that are bright on T2-weighted MRI and dark on T1-weighted MRI, liposarcomas are often difficult to distinguish from lipomas and other benign fatty tumors. Because clinical features overlap considerably between lipomas and liposarcomas, the radiographic features are of utmost importance in differentiating these entities. Key MRI features, such as homogeneity of fatty signal, absence of gadolinium enhancement, and low signal on short tau inversion recovery images, strongly support the diagnosis of benign lipoma, but the absence of these findings is not specific for liposarcoma. Unfortunately, biopsy frequently results in false negative findings; therefore, early referral to an orthopaedic oncologist is indicated when any radiographic features suggest that the mass could represent a liposarcoma. PMID- 15116656 TI - The rodded metastasis is a sarcoma: strategies to prevent inadvertent surgical procedures on primary bone malignancies. AB - Inadvertent fixation of a presumed bone metastasis may result in tumor contamination, an increased risk of local recurrence, and possibly the loss of limb or life. Although metastatic disease is far more common than primary sarcomas of bone, a rational and consistent approach is important to avoid compromising the patient's outcome. The strategy to make a diagnosis must be separate from that for fixation and reconstruction. Rarely is it possible to combine a biopsy with the reconstruction in a single procedure. Frozen section of a biopsy specimen should be performed before fixation even in patients with a previous history of carcinoma. This is especially important for the first site of osseous metastases. PMID- 15116657 TI - An approach to the management of the patient with metastatic bone disease. AB - Patients with primary sarcomas of bone and soft tissue are generally treated on protocol by a multidisciplinary team at tertiary referral centers. Conversely, patients with metastatic disease to the skeleton are often evaluated by general orthopaedic surgeons in the community. Skeletal metastases affect a growing percentage of the elderly population and can contribute to a marked decrease in their quality of life. Treatment includes stabilization with the latest prosthetic implants and new devices. Advances in the field of radiation oncology and bisphosphonate use in patients with bone metastasis are other management approaches. PMID- 15116658 TI - The art and science of reviewing manuscripts for orthopaedic journals: Part I. Defining the review. AB - The peer review process is integral to the functioning of all scientific journals and plays a pivotal role in the publication of new scientific material. Timeliness, freedom from bias, and proper review etiquette are essential for an effective review. Each type of scientific article demands a slightly different reviewing technique. Clinical research articles require that a proper question be asked and that the study methodology allows the question to be answered effectively. All studies dealing with human subjects need to be approved by the Institutional Review Board. Clinical review articles have specific criteria to determine how they should be constructed and when they should be published, whereas basic science research articles should be evaluated for their experimental method as well as the relevance of the conclusions to the data. Finally, case reports have an important place in the scientific literature, but the rationale for publishing any individual case needs to be clearly established. PMID- 15116659 TI - The art and science of reviewing manuscripts for orthopaedic journals: Part II. Optimizing the manuscript: practical hints for improving the quality of reviews. AB - Manuscripts submitted to musculoskeletal journals have several key components that need to be critically evaluated. There are specific methods to assess the abstract, illustrations, references, and other major elements of a manuscript under review. If each of these elements is assessed methodically, not only does the quality of the review improve, but it becomes more useful for the journal editor. Additionally, the method in which the review is conveyed has a marked impact on its usefulness. There should be a concise evaluation of the entire work, stating whether a publication should or should not be pursued. For poor manuscripts, several bulleted points that indicate the fatal flaw(s) are sufficient, but for good manuscripts, a systematic itemization of weaknesses will improve the quality of the manuscript. Reviews should not be derogatory and should be prompt and to the point. PMID- 15116660 TI - A registered nurse's shocking admission. PMID- 15116661 TI - Covering the uninsured. PMID- 15116662 TI - ANA fights mercury pollution. PMID- 15116663 TI - On the cutting edge. Florida nurses participate in NASA training program. PMID- 15116664 TI - Building pressure. PMID- 15116665 TI - HIV care blow. PMID- 15116666 TI - Flexible frenzy. PMID- 15116667 TI - Beating the bugs. PMID- 15116668 TI - A clean transition. PMID- 15116669 TI - In safe hands. PMID- 15116670 TI - The cost of unhealthy habits. PMID- 15116671 TI - Student Kathryn Haworth learns to cope in emergencies. PMID- 15116673 TI - A practical approach to catheter-associated problems. AB - This article examines some commonly found catheter-related problems and discusses approaches to managing and preventing them. PMID- 15116672 TI - Percutaneous vertebroplasty. AB - Percutaneous vertebroplasty is a minimally invasive procedure used to stabilise vertebral compression fractures caused by osteoporosis, haemangioma, myeloma, metastases and bone cysts. Acrylic bone cement is injected into the vertebral body to relieve pain and structurally reinforce the fracture. Interest in percutaneous vertebroplasty has grown as a result of technical procedural advances in radiology and the publication of an appraisal of, and guidelines for, the procedure by the National Institute for Clinical Excellence (2003). Nurses should be aware of the potential benefits of vertebroplasty and be involved in patient selection, and care of the patient before, during and after the procedure. Nurses should also be involved in audit analysis of the results of the procedure. More research into the effects of vertebroplasty is required and should involve nurses caring for this patient group. PMID- 15116674 TI - Stress urinary incontinence: treatment and support. AB - Stress urinary incontinence (SUI) is the involuntary leakage of urine. It is a common condition with medical and psychological consequences. Many women feel too embarrassed to seek help, therefore nurses must be aware of the symptoms and proactively identify patients. Most women with SUI would prefer to be treated in primary care settings. Specialists become involved when surgery, pelvic floor re education and drug-based treatments are used. PMID- 15116675 TI - Retrieving an assignment. PMID- 15116676 TI - Managing learning. PMID- 15116677 TI - Captive mothers. PMID- 15116679 TI - Extended independent and supplementary prescribing: an update. PMID- 15116678 TI - Skin integrity assessment in neonates and children. PMID- 15116680 TI - Planning a research project. PMID- 15116681 TI - Understanding smoking behaviour in children and adolescents. PMID- 15116682 TI - Implementing equipment guidance in the community. PMID- 15116683 TI - Adolescents with diabetes: a health action zone project. PMID- 15116684 TI - Pain assessment in children with neurological impairment. PMID- 15116685 TI - Clinical leaders in paediatric nursing: a pilot study. AB - This pilot study explored the appropriateness and suitability of a questionnaire for discovering who the clinical nurse leaders are in a paediatric unit of a large NHS trust and what the qualities and characteristics of clinical leaders might be. Thirteen respondents made 24 separate nominations for clinical leaders, 20 of whom were 'F' grade junior sisters who were seen as having the clinical leadership qualities of: coping well with change; integrity; being supportive; considering relationships valuable; being flexible; and clinical competence. These results highlight aspects of clinical leadership in paediatrics and the value of conducting a pilot study. PMID- 15116686 TI - Origin and organization of brainstem catecholamine innervation in the rat. AB - The catecholamine (CA) innervation of the rat brainstem was studied by biochemical analysis of discrete nuclei or areas and by glyoxylic acid formaldehyde freeze dry fluorescence histochemistry. CA assays demonstrate that the highest norepinephrine (NE) content in brainstem is present in the trigeminal motor nucleus, nucleus tractus solitarius, dorsal motor nucleus of the vagus and nucleus raphe dorsalis. Bilateral locus coeruleus (LC) lesions do not significantly alter NE content in these nuclei but do decrease NE content in the superior and inferior colliculi, medial geniculate body, interpeduncular nucleus, pontine nuclei and the main sensory trigeminal nucleus (60-75%). Dopamine (DA) and epinephrine (E) are found in significant concentration in only a few of the nuclei examined. Fluorescence histochemical analysis indicates that two groups of NE axons innervate rat brainstem. LC neuron axons with a distinctive morphology principally innervate sensory and association nuclei of the brainstem. These disappear completely after bilateral LC lesions. The second group of axons originates from lateral and dorsal tegmental NE cell groups. Primary motor and visceral nuclei are densely innervated by fine and thick axons from these groups. Lesions of LC do not alter the NE innervation in any of the nuclei which contain axons of the second group. These results indicate that the brainstem NE innervation is divided into two major systems. The locus coeruleus complex innervates mainly primary sensory and association nuclei whereas the lateral tegmental NE neurons innervate primary motor and visceral nuclei. Although some overlap is present, the LC and lateral tegmental NE systems predominantly innervate separate and functionally distinct areas of the brainstem. DA and E neurons provide a very minor component of the brainstem CA innervation. PMID- 15116687 TI - Pikeperch horizontal cells identified by intracellular staining. AB - Micropipettes filled with Procion yellow dye were used to record from and to stain pikeperch horizontal cells intracellularly. Three major types were found: a distal layer (H1) of relatively small cells which were luminosity or L-type; a second and more proximal layer of larger L-type cells (H2); and a third and yet more proximal layer of very stellate chromatic or C-type cells (H3). A few anucleate processes which displayed slow-potentials were found in the proximal area of the inner nuclear layer. Cells of each of the three layers were shown to be cone related by both anatomical and physiological methods. L-type cells were further categorized by the area over which each exhibited spatial summation and the relative sensitivity of each to red and green lights. Receptive field sizes of H2's were found to range from less than 2 mm to greater than 5 mm in diameter, whereas those of the few H1's tested were all less than 2 mm. Results from spectral screening tests indicate that most H1 and H2 cells are maximally sensitive to orange light, whereas the H3 cells hyperpolarize maximally to green and depolarize maximally to red. A small percentage of sampled C-cells displayed an additional depolarization to violet. PMID- 15116688 TI - Synaptic connections linking cones and horizontal cells in the retina of the pikeperch (Stizostedion vitreum). AB - Cones and horizontal cells of the pikeperch retina were studied with morphological and physiological techniques. Gap junctions were observed between cone pedicles and basal processes emitted by neighboring cones. Intracellular recordings showed that the light-evoked hyperpolarizing cone response was enhanced by light falling upon neighboring receptors within a radius of 50 microns. We suggest that the network of gap junctions between cones mediates the summative lateral interaction described. Three sub-classes of horizontal cells (H1, H2, H3) send dendrites to cones; H1 and H2 cells appear to contact twin cones, exclusively or preferentially, whereas H3 cells appear to synapse only with single cones. Horizontal cells of the same sub-class are joined by gap junctions between dendrites or at the lateral faces of perikarya. These unions extend over several micron 2 and as seen in transmission electron microscopy consist of patches of close apposition alternating with areas of membrane separation, folding and occasional zonulae adherents. Freeze-fracture profiles of horizontal gap junctions show localized areas of dense particle aggregation on the P-face and pits on the E-face flanked by regions of unspecialized membrane. These morphological findings provide support for the known spatial and color coding properties of pikeperch horizontal cells. PMID- 15116689 TI - An autoradiographic study of the retinotectal projection in the golden hamster. AB - Intraocular injections of tritiated leucine and proline were used to examine the retinotectal projection of the golden hamster. In the contralateral superior colliculus intense and complete label was seen in the stratum zonale, stratum griseum superficiale and the upper portion of the stratum opticum, with relatively less dense label in the lower part of the optic layer. On the ipsilateral side no label was found in the most rostral portion of the tectum. This area comprised about 10% of the rostro-caudal extent of the colliculus, and most likely, it receives a crossed input from the temporal retina (as demonstrated in the cat by Harting and Guillery, '76). Very sparse label was observed in an anterior segment of the ipsilateral colliculus. In coronal sections it appeared as discrete clumps or patches which were confined to the stratum opticum. Within this layer there was a tendency for the clumps to be located more dorsally with increasing laterality. There was considerable variability between and within animals in the size of the clumps as well as the distance between clumps. Reconstruction of coronal sections showed that the ipsilateral label forms discontinuous ribbons which extend up to 180 microns in the rostro-caudal dimension. No label was seen on the ipsilateral side in the remaining tectum (caudal 60%). PMID- 15116690 TI - Membrane specializations and cytoplasmic channels of Schwann cells in mammalian peripheral nerve as seen in freeze-fracture replicas. AB - Mammalian Schwann cells in rat, rabbit and human fetal nerves were studied using several cryoprotective agents for electron microscopic study of freeze-fracture replicas. The findings in fixed and unfixed tissue reveal surface plasmalemma caveolar specializations and the outer layer membrane junctional complexes found in non-mammalian species. The plasmalemma also reveals a complex arrangement of contours outlining cytoplasmic channel networks distinct from the long-recognized Schmidt-Lanterman incisures and paranodal cytoplasmic loops. A specialized interconnected channel system in the outer "loose" myelin layer displays relatively uniform dimensions comparable in diameter to nodal microvilli, paranodal loops and some incisures. An adaxonal tubular channel system constituting the "axon-Schwann network" is found in the internodal region in addition to other variants of the adaxonal Schwann plasmalemma. The several forms of sequestration of Schwann cell cytoplasm presumably underlie the specialized needs of cytoplasmic continuity in a dynamic functional entity in which large domains of cytoplasm have been displaced by the formation of compact myelin. PMID- 15116691 TI - A golgi study of the class V cell in the visual thalamus of the cat. AB - Golgi methods were used to study class V cells within the cat visual thalamus. Counterstaining was combined with Golgi staining to assess the distribution of dendrites relative to cytoarchitectural boundaries. Class V cells were encountered within all laminae of the lateral geniculate nucleus, the medial interlaminar nucleus, and the lateral posterior complex. The cells possess medium sized perikarya and smooth and varicose or moniliform dendrites. Dendritic appendages are sparse and occur as single or serial swellings on thin processes. Many class V cells exhibit large, sparse dendritic arbors which span laminar or nuclear borders; dendrites were seen to lie within and to cross the interlaminar zones of the lateral geniculate nucleus, and extend beyond this nucleus into the perigeniculate nucleus and medial interlaminar nucleus. Class V cells of the lateral posterior complex send dendrites into the external medullary lamina. Indirect evidence favors the interpretation that the class V cells are thalamo cortical relay cells. PMID- 15116692 TI - The connections of the septal region in the rat. AB - The efferent, afferent and intrinsic connections of the septal region have been analyzed in the rat with the autoradiographic method. The lateral septal nucleus, which can be divided into dorsal, intermediate and ventral parts, receives its major input from the hippocampal formation and projects to the medial septal diagonal band complex. The ventral part of the nucleus also sends fibers through the medial forebrain bundle to the medial preoptic and anterior hypothalamic areas, to the lateral hypothalamic area and the dorsomedial nucleus, to the mammillary body (including the supramammillary region), and to the ventral tegmental area. The medial septal nucleus/diagonal band complex projects back to the hippocampal formation by way of the dorsal fornix, fimbria, and possibly the cingulum. Both nuclei also project through the medial forebrain bundle to the medial and lateral preoptic areas, to the lateral hypothalamic area, and to the mammillary complex. The medial septal nucleus also sends fibers to the midbrain (the ventral tegmental area and raphe nuclei) and to the parataenial nucleus of the thalamus, while the nucleus of the diagonal band has an additional projection to the anterior limbic area. Ascending inputs to the medial septal nucleus/diagonal band complex arise in several hypothalamic nuclei and in the brainstem aminergic cell groups. The posterior septal nuclei (the septofimbrial and triangular nuclei) receive their major input from the hippocampal formation, and project in a topographically ordered manner upon the habenular nuclei and the interpeduncular nuclear complex. The bed nucleus of the stria terminalis receives its major input from the amygdala (Krettek and Price, '78); but other afferents arise from the ventral subiculum, the ventromedial nucleus, and the brainstem aminergic cell groups. The principal output of the bed nucleus is through the medial forebrain bundle to the substantia innominata, the nucleus accumbens, most parts of the hypothalamus and the preoptic area, the central tegmental fields of the midbrain, the ventral tegmental area, the dorsal and median nuclei of the raphe, and the locus coeruleus. The bed nucleus also projects to the anterior nuclei of the thalamus, the parataenial and paraventricular nuclei, and the medial habenular nucleus, and through the stria terminalis to the medial and central nuclei of the amygdala, and to the amygdalo-hippocampal transition area. PMID- 15116693 TI - Lessons from the 11th conference on retroviruses and opportunistic infections. PMID- 15116694 TI - Two patients with a similar non-AIDS-defining malignancy. PMID- 15116695 TI - Pegasys-Copegus combination treats both HIV and hepatitis C. PMID- 15116696 TI - Marijuana eases HIV-related nerve pain. PMID- 15116697 TI - Antiretroviral simplification: multiple benefits, some concerns. PMID- 15116698 TI - HIV-associated musculoskeletal involvement. AB - Various rheumatic manifestations associated with HIV infection have been recognized, ranging from infectious conditions, such as septic arthritis, to seronegative spondyloarthropathy. Other musculoskeletal manifestations include lupus-like and Sjogren-like diseases and HIV-related malignancy. The introduction of HAART has changed the spectrum of the clinical manifestations of rheumatic disease seen today, with infections and articular involvement being the most frequently observed. PMID- 15116699 TI - Editorial comment: HIV-1 infection and the rheumatic diseases--new issues, new challenges, same old disparities. PMID- 15116700 TI - Effects of adjuvant radiotherapy for testicular cancer on CD4+ cell count in HIV positive patients: a case report. AB - The occurrence of testicular seminoma is more frequent in HIV-positive than in HIV-negative men. Management involves orchiectomy and radiotherapy for early stage seminoma. Radiotherapy is typically well tolerated, with minimal side effects. This Case Report presents an HIV-positive patient who experienced a decrease in CD4+ cell count and an alteration of his clinical HIV disease course following radiotherapy for seminoma. PMID- 15116701 TI - Editorial comment: adjuvant radiotherapy for testicular cancer--balancing cancer treatment with HIV management. PMID- 15116702 TI - Optimal delivery of HAART during hospitalization. PMID- 15116703 TI - Inherited ossifying diseases. AB - Inherited ossifying diseases are relatively uncommon diseases leading ta a great disability and life-threatening complications. Fibrodysplasia Ossificans Progressiva is characterized by the association of skeletal abnormalities mainly in great toes, and enchondral ossifications in tendons and muscles. BMP dysregulation seems to be the main underlying mechanism of the heterotopic ossifications. The genetic basis remain controversial between a mutation on chromosome 4 or 17. Progressive Osseous Heteroplasia (HOP), more recently described, shares some similarities with Albrights hereditary osteodystrophy. In HOP, the intramembranous ossifications progressively developped from the dermis to the deeper layer. The genetic abnormality involved the GNAS 1 gene leading to an inactivation of the alpha subunit of the G protein-complex. Some therapeutic approaches have been tried: angiogenesis inhibition, mast cell inhibition; others remained in project: BMP 4 inhibition; actually there is no proved efficacy of any of them. PMID- 15116704 TI - Osteoporosis and breast cancer. AB - Osteoporosis affects one in three women after the menopause and the incidence of osteoporotic fractures increases steadily throughout life. Breast cancer is the most common cancer in women, both before and after the menopause. In younger women, recovery from breast cancer has been achieved using aggressive chemotherapy and radiotherapy that can adversely affect bone tissue or induce premature menopause. In postmenopausal women, breast cancer and osteoporosis are common, and although both are dependent on estrogens this leads to conflicting implications for the diagnosis and treatment: estrogens reduce the risk of fractures but increase the risk of breast cancer. Estrogen supplementation is, therefore, contraindicated in patients with a history of breast cancer. Selective estrogen response modifiers (SERMs) hold great promise, as they decrease both the fracture risk via an estrogen-agonist effect on bone and the breast cancer risk via an estrogen-antagonist effect on the breast tissue. SERMs can be used after successful treatment for breast cancer. Bisphosphonates, which are potent bone resorption inhibitors, are widely used both in cancer patients and in the prevention and treatment of spinal and peripheral osteoporotic fractures. Contraindications are exceedingly rare, and the satisfactory safety profile of these agents can be expected to improve further with newly developed modes of administration. Whether the bisphosphonates currently used to treat osteoporosis (alendronate and risendronate) have beneficial effects on skeletal events related to cancer progression remains to be determined, however. In sum, selection of the optimal treatment for osteoporosis in a patient with breast cancer involves assessment of the risk/benefit ratio of each treatment option, based on patient age, other risk factors for osteoporosis, and the stage of breast cancer progression. PMID- 15116705 TI - Management of primary hyperthyroidism: toward minimal access surgery. AB - Fifteen years ago, bilateral exploration of the neck was dogma in parathyroid surgery. Now, less invasive procedures can be used to target lesions identified by new tests such as dual-phase Sestamibi scanning or intraoperative documentation of parathyroid hormone (PTH) level changes after removal of a parathyroid gland. A hand-held gamma probe can be used for intraoperative detection of high-uptake lesions, and video-assisted endoscopic surgery has been used successfully. With these new techniques, surgical exploration can be confined to one side of the neck through smaller incisions associated with better cosmetic results. The operating time is reduced, and in some cases the procedure can be done under local anesthesia. The objective of this article is to describe recent changes in the management of parathyroid adenoma requiring surgery. PMID- 15116706 TI - Prairie dog: cuddly pet or Trojan horse? PMID- 15116707 TI - Early defervescence and SARS recovery. PMID- 15116708 TI - Babesiosis in Fairfield County, Connecticut. PMID- 15116709 TI - Migratory thrombophlebitis and acute Q fever. PMID- 15116710 TI - Report of the activities of the American Board of Surgery (ABS) to the Association of Program Directors in Surgery (APDS). PMID- 15116711 TI - Residency Review Committee (RRC) report. PMID- 15116712 TI - West Nile poliomyelitis. PMID- 15116714 TI - Schistosoma haematobium infection and Buruli ulcer. PMID- 15116713 TI - Typhus group Rickettsiae antibodies in rural Mexico. PMID- 15116715 TI - 1998 dengue hemorrhagic fever epidemic in Taiwan. PMID- 15116716 TI - Rift Valley fever encephalitis. PMID- 15116717 TI - Anthrax toxin: can a little be a good thing? PMID- 15116718 TI - The development of Ca2+ indicators: a breakthrough in pharmacological research. AB - The development, beginning in 1979, of fluorescent Ca2+-specific indicators as research tools has revolutionized transmembrane signaling studies. In this article, the state of the art in the 'pre-Ca2+-indicator' era and the rationale for the development of indicators trapped in the cytosol to investigate the cytosolic concentration of Ca2+ in mammalian cells are summarized. Subsequent extension of these studies to the level of the single cell, together with the unique impact that Ca2+ indicators have had on signaling research and the introduction of specific, fluorescent gene constructs that provide direct, high resolution information about the intracellular concentration of Ca2+, are also discussed. PMID- 15116719 TI - Diacylglycerol's affair with protein kinase C turns 25. AB - This year marks the 25-year anniversary of the discovery by Nishizuka and co workers that diacylglycerol activates the ubiquitous signal transducer protein kinase C. This discovery placed the lipid second messenger-protein kinase C signaling pathway center stage alongside the cAMP-protein kinase A pathway, which was already established as a fundamental mechanism for transducing extracellular signals. PMID- 15116720 TI - HIV-1 Vif versus APOBEC3G: newly appreciated warriors in the ancient battle between virus and host. PMID- 15116721 TI - p53: 25 years after its discovery. AB - Since its discovery 25 years ago, the p53 protein has emerged as a key tumor suppressor protein at the crossroads of cellular stress response pathways. Through these pathways, which can lead to cell-cycle arrest, DNA repair, cellular senescence, differentiation and apoptosis, p53 facilitates the repair and survival of damaged cells or eliminates severely damaged cells from the replicative pool to protect the organism. Because of these dynamic and multiple functions of p53, which are largely lost following mutations in the gene encoding p53, this molecule continues to be studied intensively in biomedical research, including the fields of toxicology and pharmacology. In this article, we briefly review the first 25 years of research on p53. PMID- 15116722 TI - Gene duplication and biased functional retention of paralogs in bacterial genomes. PMID- 15116723 TI - Bugs N the 'hood. PMID- 15116724 TI - Market-driven department benchmarks. PMID- 15116725 TI - [Better care provided for adults with congenital heart defects]. PMID- 15116726 TI - Special issue dedicated to the Memory of Dr. Donald Reis. PMID- 15116727 TI - Controversy on adult stem-cell plasticity and rules governing the use of fetal tissue in China. PMID- 15116728 TI - [Life style and cost explosion in health care]. PMID- 15116729 TI - [Life style and cost explosion in health care]. PMID- 15116730 TI - [Life pictures. Ursula Ravens]. PMID- 15116731 TI - Proceedings of the Sixth International Congress on Pediatric Pulmonology. February 28-March 2, 2004. Lisbon, Portugal. PMID- 15116732 TI - Soybeans. PMID- 15116733 TI - Teamwork. PMID- 15116734 TI - Abstracts from the 12th European Workshop on Neonatology. PMID- 15116735 TI - Process validation for manufacturing of biologics and biotechnology products. Proceedings of a meeting. September 6-7, 2001. Berlin, Germany. PMID- 15116736 TI - Selective COX-2 inhibition endothelial function in coronary artery disease. Chenevard R, Hurlimann D, Bechir M et al Circulation 2003; 107: 405-09. PMID- 15116737 TI - Short reviews and abstracts from the 11th International Vasculitis and ANCA Workshop. October 2-5, 2003, Prague, Czech Republic. PMID- 15116738 TI - New genetic and metabolic insights into animal models of diabetes. Proceedings of the 9th International Workshop on Lessons from Animal Diabetes. June 17-21, 2003. Bar Harbor, Maine, USA. PMID- 15116739 TI - Mortality trends of rectal cancer in Japan: 1960-2000. PMID- 15116740 TI - Hepatitis C virus ultrastructure and morphogenesis. AB - Details of the ultrastructure of hepatitis C virus (HVC) virion remain unclear because it has proved extremely difficult to visualise virus particles from infected serum and tissues directly. In addition, although much is known about the viral genome, first cloned in 1989, little is known about HCV morphogenesis, due to the lack of an efficient in vitro culture system for HCV propagation. Virus-like particles (VLPs) obtained by expressing genes encoding the HCV structural proteins in mammalian cells can be used as an alternative model for studying HCV morphogenesis. In particular, this HCV-LP model has made it possible to demonstrate that HCV budding occurs at the ER membrane and that the core protein drives this process. The HCV-LP model opens up new possibilities for the investigation of viral morphogenesis and virus-host cell interactions, which may make it possible to establish the long-awaited in vitro culture system for HCV. PMID- 15116741 TI - [Ultrasound and enchondral osteogenesis: an experimental study]. PMID- 15116742 TI - [Dynamics of acid base balance in the oral cavity of patients with orthodontic constructions]. PMID- 15116744 TI - [Clinical morphological characteristics of changes in the minor salivary glands of tobacco smokers]. PMID- 15116743 TI - Cardiovascular fitness and neurocognitive function in older adults: a brief review. AB - We provide a brief review of the extant research on the influence of cardiovascular fitness training on brain and cognition. The review includes an examination of the non-human animal literature that has reported molecular, cellular, and behavioral consequences of fitness interventions. We relate this literature to human studies of the relationship between fitness and cognition, as well as the nascent literature on fitness influences on human brain structure and function with state-of-the art neuroimaging techniques. We also consider the important topic of participant adherence in clinical exercise trials. Finally, we suggest future directions for studies of cardiovascular fitness, aging, and neurocognitive function. PMID- 15116745 TI - [Long-term results of vital and devital pulpectomy]. PMID- 15116746 TI - [Results of treatment of patients with pericoronaritis depending on the disease form and treatment method]. PMID- 15116747 TI - [Stages in repair of maxillodental deformations in combined therapy of congenital facial clefts]. PMID- 15116748 TI - Software supplier guide. PMID- 15116749 TI - [Osteoplastic operations in combined treatment of maxillary deformations in congenital cleft of the upper lip]. PMID- 15116750 TI - [Quantitative physico-chemical characteristics of the oral liquid in preschool children]. PMID- 15116751 TI - [Role of Professor R.D. Novoselov in the development of childhood dentistry]. PMID- 15116752 TI - [Contribution of the stomatology department of the Tver State Medical Academy to the development of stomatological science in Russia]. PMID- 15116753 TI - [The research progress of fibronectin EDA's expression and functions]. PMID- 15116754 TI - Migration of substances from food packaging materials to foods. AB - The employment of novel food packaging materials has increased the number of occurring hazards due to the migration from packaging material to the packaged food. Although polymers have mainly monopolized the interest of migration testing and experimentation, recent studies have revealed that migration also occurs from "traditional" materials generally considered to be safe, such as paper, carton, wood, ceramic, and metal. The regulations and the directives of the EU tend to become stricter in this respect. The emphasis is on reaching a consensus in terms of food simulants and testing conditions for migration studies. Furthermore, the list of hazardous monomers, oligomers, and additives continues to augment in order to ensure that the consumer safety is in current agreement with the HACCP, which is continuously gaining ground. PMID- 15116755 TI - Useful byproducts from cellulosic wastes of agriculture and food industry--a critical appraisal. AB - Cellulose, an important cell wall polysaccharide, which is replenished constantly in nature by photosynthesis, goes waste in a lion's share in the form of pre harvest and post-harvest agricultural losses and wastes of food processing industry. These cellulose wastes have an immense potential to be utilized for the production and recovery of several products and ingredients in food application. In this present study, a wide spectrum of researches in the arena of properties of cellulose, hemicellulose and lignin; their degradation; sources and composition of cellulosic and lignocellulosic wastes of agriculture and food industry; present status of converting them into value-added products of food applications; constraints in their conversions and future prospects therein has been reviewed in details. The study has encompassed production of biomass for various utilization and production and recovery of protein and amino acids, carbohydrates, lipids, organic acids, foods & feeds and other miscellaneous products. PMID- 15116756 TI - Aloe vera: a valuable ingredient for the food, pharmaceutical and cosmetic industries--a review. AB - Scientific investigations on Aloe vera have gained more attention over the last several decades due to its reputable medicinal properties. Some publications have appeared in reputable Scientific Journals that have made appreciable contributions to the discovery of the functions and utilizations of Aloe- "nature's gift." Chemical analysis reveals that Aloe vera contains various carbohydrate polymers, notably glucomannans, along with a range of other organic and inorganic components. Although many physiological properties of Aloe vera have been described, it still remains uncertain as to which of the component(s) is responsible for these physiological properties. Further research needs to be done to unravel the myth surrounding the biological activities and the functional properties of A. vera. Appropriate processing techniques should be employed during the stabilization of the gel in order to affect and extend its field of utilization. PMID- 15116757 TI - Biological properties of curcumin-cellular and molecular mechanisms of action. AB - Curcuminoids, a group of phenolic compounds isolated from the roots of Curcuma longa (Zingiberaceae), exhibit a variety of beneficial effects on health and on events that help in preventing certain diseases. A vast majority of these studies were carried out with curcumin (diferuloyl methane), which is a major curcuminoid. The most detailed studies using curcumin include anti-inflammatory, antioxidant, anticarcinogenic, antiviral, and antiinfectious activities. In addition, the wound healing and detoxifying properties of curcumin have also received considerable attention. As a result of extensive research on the therapeutic properties of curcumin, some understanding on the cellular, molecular, and biochemical mechanism of action of curcumin is emerging. These findings are summarized in this review. PMID- 15116758 TI - The relative effect of milk base, starter, and process on yogurt texture: a review. AB - Yogurt is a milk curd produced all over the world, obtained by a lactic fermentation of a milk base enriched with milk proteins, and sometimes sugars and thickeners. One of the most important sensorial attributes for yogurt is texture, which could be assessed by sensory or instrumental analysis. A lot of work has been published in studying the contribution of milk base, starter, and process on yogurt texture in order to develop new textures, or simply to reduce fat content, or the level of addition of protein and thickener in milk. However, these studies are limited to only a few factors. The topic of this review was to synthesize the data of literature, with the aim of extracting and classifying factors on the basis of their influence on yogurt texture. Three factors, milk base heating, starter, and yogurt shearing after fermentation, respectively, play a key role in the elaboration of texture. The control of these three parameters allows the improvement of the textural attributes of yogurts by 2 to 15 times. PMID- 15116759 TI - Catastrophe and what to do about it if you are a bacterium: the importance of frameshift mutants. AB - Key problems that bacteria have historically faced are the challenges of the lack of essential nutrients and the presence of antibiotics produced naturally, but there are many other challenges. It appears that for many of these challenges the bacteria have mechanisms encoded in their genomes that are not usually functioning, but may be "turned on" when needed, even if the need only occurs once in hundreds of thousands of generations. Such mechanisms at other times somehow need to be "turned off" because they may cause a slight disadvantage, or even a grave disadvantage to the cell compared with wild-type cells during the time the population is not being challenged. On the other hand, a gene cannot simply be discarded because it might be needed again. How do microorganisms solve the problem of responding to challenges that only occur rarely? I suggest that in most cases, the mutation must occur by the existence of a readily reversible mutation. The mutation in likely the result of a frameshift mutation that caused the response and later another frameshift occurs to return the genome to its original state. PMID- 15116760 TI - Bacterial separation and concentration from complex sample matrices: a review. AB - The use of many rapid detection technologies could be expanded if the bacteria were separated, concentrated, and purified from the sample matrix before detection. Specific advantages of bacterial concentration might include facilitating the detection of multiple bacterial strains; removal of matrix associated assay inhibitors; and provision of adequate sample size reduction to allow for the use of representative food sample sizes and/or small media volumes. Furthermore, bacterial concentration could aid in improving sampling techniques needed to detect low levels of pathogens or sporadic contamination, which may perhaps reduce or even eliminate the need for cultural enrichment prior to detection. Although bacterial concentration methods such as centrifugation, filtration, and immunomagnetic separation have been reported for food systems, none of these is ideal and in many cases a technique optimized for one food system or microorganism is not readily adaptable to others. Indeed, the separation and subsequent concentration of bacterial cells from a food sample during sample preparation continues to be a stumbling block in the advancement of molecular methods for the detection of foodborne pathogens. The purpose of this review is to provide a detailed understanding of the science, possibilities, and limitations of separating and concentrating bacterial cells from the food matrix in an effort to further improve our ability to harness molecular methods for the rapid detection of foodborne pathogens. PMID- 15116761 TI - Extended-spectrum beta-lactamases (ESbLs): characterization, epidemiology and detection. AB - Beta-lactamases of Enterobacteriaceae are the most important mechanism of resistance against beta-lactam drugs. Two types of beta-lactamases can confer resistance against 3rd generation cephalosporins. Chromosomally mediated beta lactamases are inducible and are not inhibited by clavulanic acid. Resistance due to these enzymes is non-transferable. The 2nd type of enzyme is plasmid-mediated beta-lactamases, which are inhibited by clavulanic acid. These enzymes are more important clinically as these can be transferred between various species of Enterobacteriaceae. These enzymes are called extended-spectrum beta-lactamases (ESBLs). ESBL-producing Enterobacteriaceae have been responsible for numerous outbreaks of infection throughout the world and pose challenging infection control issues. Antibacterial choice is often complicated by multi-resistance. ESBLs can confer resistance against all beta-lactam drugs except carbapenems and cephamycins. Nursing home patients may be an important reservoir of ESBL containing multiple antibiotic-resistant organisms. Use of broad-spectrum oral antibiotics and probably poor infection control practices may facilitate spread of this plasmid-mediated resistance. In addition to known populations at risk, ambulatory patients with chronic conditions represent another patient population that may harbor ESBL-producing organisms. Various methods can be used for detection of ESBLs in the laboratory. These tests include double disc diffusion test, Vitek ESBL test, E Tests, MIC Determination, genetic method, and isoelectric focusing (IEF). PMID- 15116762 TI - Bacterial insecticidal toxins. AB - Over the years it has been important for humans to control the populations of harmful insects and insecticides have been used for this purpose in agricultural and horticultural sectors. Synthetic insecticides, owing to their various side effects, have been widely replaced by biological insecticides. In this review we attempt to describe three bacterial species that are known to produce insecticidal toxins of tremendous biotechnological, agricultural, and economic importance. Bacillus thuringiensis (BT) accounts for 90% of the bioinsecticide market and it produces insecticidal toxins referred to as delta endotoxins. The other two bacteria belong to the genera Photorhabdus and Xenorhabdus, which are symbiotically associated with entomopathogenic nematodes of the families Heterorhabditidae and Steinernematidae respectively. Whereas, Xenorhabdus and Photorhabdus exist in a mutualistic association with the entomopathogenic nematodes, BT act alone. BT formulations are widely used in the field against insects; however, over the years there has been a gradual development of insect resistance against BT toxins. No resistance against Xenorhabdus or Photorhabdus has been reported to date. More recently BT transgenic crops have been prepared; however, there are growing concerns about the safety of these genetically modified crops. Nematodal formulations are also used in the field to curb harmful insect populations. Resistance development to entomopathogenic nematodes is unlikely due to the physical macroscopic nature of infection. Xenorhabdus and Photorhabdus transgenes have not yet been prepared; but are predicted to be available in the near future. In this review we start with an overview of the synthetic insecticides and then discuss Bacillus thuringiensis, Xenorhabdus nematophilus, and Photorhabdus luminescens in greater detail. PMID- 15116763 TI - [Friendliness and quality assurance]. PMID- 15116764 TI - [Bacterial meningitis]. PMID- 15116765 TI - [Epidemiology and prevention of meningococcal illnesses--Europe, Germany]. PMID- 15116766 TI - [Vaccination--myths and reality]. PMID- 15116767 TI - [Tuberculosis--the 3rd most frequent infectious disease for health care personnel]. PMID- 15116769 TI - [Therapy of advanced ovarian carcinoma]. PMID- 15116768 TI - [Pegvisomant decreases growth hormone to nothing]. PMID- 15116770 TI - [Recent progress in therapy of gastrointestinal tumors]. PMID- 15116771 TI - [Better quality of life for tumor patients]. PMID- 15116772 TI - [Tick-borne meningoencephalitis at maximum prevalence]. PMID- 15116773 TI - [Importance of early diagnosis and therapy]. PMID- 15116774 TI - [Rapid hyposensitization]. PMID- 15116775 TI - [Obesity]. PMID- 15116777 TI - [Risk of infection is underestimated]. PMID- 15116776 TI - [Gentle motion to control decubitus ulcer]. PMID- 15116778 TI - [One more good resolution...]. PMID- 15116779 TI - [Miele disinfection OxiVario Plus equipment removes the last protein residues]. PMID- 15116780 TI - [Recognition of involvement to benefit schizophrenic patients]. PMID- 15116781 TI - [Depression in the elderly--frequently this treatable illness is not recognized]. PMID- 15116782 TI - [Success with engaged educational work]. PMID- 15116783 TI - [Better coordination of patient care by nurses and physicians]. PMID- 15116784 TI - [A helping hand for family caregivers]. PMID- 15116785 TI - [Medical coding assistants]. PMID- 15116786 TI - [Women and (secret) addiction (1)]. PMID- 15116787 TI - [Interview with Rolf Hullenhorst, director of the German Central Department for Addiction Questions e.V., Hamm]. PMID- 15116789 TI - [What can one do in such a case?]. PMID- 15116790 TI - Alpha and omega of microbes. Antibiotics and probiotics: judicious use is the key. PMID- 15116791 TI - Specialists really do need family physicians. PMID- 15116792 TI - Charting made easier and faster. PMID- 15116793 TI - Summarizing ordinal data. What is appropriate? PMID- 15116794 TI - New guidelines on concussion management overlooked. PMID- 15116795 TI - New guidelines on concussion management overlooked. PMID- 15116796 TI - Debating the values of family medicine. PMID- 15116797 TI - Prescriptions of sleep. PMID- 15116799 TI - Dermacase. Allergic contact dermatitis. PMID- 15116798 TI - Hypothyroidism during pregnancy. AB - QUESTION: I have a 27-year-old patient diagnosed with hypothyroidism in the 8th week of pregnancy. She received conflicting opinions regarding risk for her baby and wants to get more information. I also found conflicting information in the literature. How should I advise her? ANSWER: Pregnant patients with untreated hypothyroidism are at increased risk of obstetric complications. Adequate treatment with thyroid hormones greatly reduces the frequency of these complications. Observational studies suggest that children whose mothers had hypothyroxinemia in early pregnancy have lower IQs than matched controls. Another study has shown that even if levothyroxine therapy is started after the first trimester, there is an excellent chance children will have normal neuropsychologic development. PMID- 15116800 TI - Practice tips. Colorectal cancer screening in my practice. PMID- 15116801 TI - Approach to managing diabetic foot ulcers. AB - INTRODUCTION: Of an estimated 1.7 to 2 million Canadians with diabetes, approximately 10% will present each year to their family doctors with plantar ulcers. Nearly 3500 will require major lower extremity amputations. SOURCES OF INFORMATION: Most of the recommendations outlined in this paper are based on level I evidence from excellent bench research and epidemiologic studies. MAIN MESSAGE: Both insulin-dependent and non-insulin-dependent diabetics develop foot infections. These patients are on average 60 years old and have had diabetes for more than 10 years. Physicians who insist on excellent blood sugar control, provide ongoing patient education on diabetic foot care, prescribe appropriate shoes, and practise an aggressive multidisciplinary approach to wound care can reduce the rate of lower extremity amputations by more than 50%. CONCLUSION: Foot problems remain one of the main challenges associated with diabetes, but family physicians can manage them successfully. PMID- 15116802 TI - Antibiotics for lower respiratory tract infections. Still too frequently prescribed? AB - OBJECTIVE: To examine how Canadian family physicians currently prescribe for lower respiratory tract infections (LRTIs). DESIGN: Prospective assessment of adults with symptoms of LRTIs. SETTING: Offices of 120 community-based members of the College of Family Physicians of Canada. PARTICIPANTS: Four hundred seven adults (16 years and older). MAIN OUTCOME MEASURES: Clinical findings, diagnoses, tests ordered, and prescriptions for antibiotics were documented on a standardized form. RESULTS: Antibiotics were prescribed to 58.4% of patients presenting with symptoms of LRTIs. Prescribing was higher (77.9%) for those diagnosed with acute bronchitis, which accounted for 70.3% of prescriptions. Physicians were often uncertain about the need for antibiotics, were concerned that patients could become sicker, and felt pressured by patients to prescribe antibiotics. Macrolides were most frequently prescribed; no tests were ordered in 85.0% of encounters. CONCLUSION: The number of antibiotic prescriptions for adults with LRTIs remains high in Canada. Rates of prescribing are increased by diagnosis of acute bronchitis, clinical uncertainty, pressure from patients to receive antibiotics, and concern that patients will deteriorate if left untreated. PMID- 15116804 TI - Caveat emptor. "Probiotics" might not be what they seem. AB - OBJECTIVE: To assess whether commercially prepared probiotic products contain viable organisms, as claimed by their manufacturers, particularly whether products labeled as containing Lactobacillus did so. To identify and quantify as many species as feasible and to compare them with the contents listed on labels. DESIGN: Randomized, double-blind trial. SETTING: Community hospital in Chilliwack, BC. MAIN OUTCOME MEASURES: Trial of 10 randomly chosen brands of probiotic preparations bought over-the-counter in British Columbia's lower mainland. Only products claiming to contain lactobacillus were included in this study. Viable organisms in each probiotic brand and quantities of Lactobacillus in each product. RESULTS: None of the 10 products tested matched their labeled microbiologic specifications. Two brands grew nothing aerobically or anaerobically. No Lactobacillus grew in five brands, although their labels stated that this was the main species. Eight brands contained viable cells, but only 10% of the number stated by their manufacturers. CONCLUSION: Most product labels did not adequately identify or quantify microbes. Use of probiotics should not be recommended at this time. PMID- 15116803 TI - Short report: Ceftriaxone for cat and dog bites. Simple outpatient treatment. PMID- 15116805 TI - 2003 Canadian hypertension recommendations. It's not all old hat. PMID- 15116806 TI - Colorectal cancer. Summary of Canadian screening guidelines. PMID- 15116807 TI - Which hand-held computer is better for doctors? Part 2: Comparing models with Microsoft operating systems. PMID- 15116808 TI - Moving our children to health. Active play every day. PMID- 15116809 TI - You and the college. PMID- 15116810 TI - Physicians at the bedside. Family physicians caring for hospital inpatients. PMID- 15116811 TI - Ghostwriters for doctoral thesis. PMID- 15116812 TI - Quantitative and qualitative aspects of diagnosing acute appendicitis. PMID- 15116813 TI - Terror bombing-mechanisms, consequences and implications. AB - Our objective in this review is to describe the unique features of bombing injury and to outline some special elements of their management. This is to allow the timely improvement and adjustment of existing mass casualty protocols. Forensic studies, detonation and explosion, mechanisms of injury in explosion and their bodily effects, chemical effects of the explosive, site of the explosion and the wounding potential, the Multidimensional Injury Pattern, diagnostic evaluation of Multidimensional Injury Pattern, and surgical and treatment dilemmas associated with it are described and discussed. PMID- 15116814 TI - The potential use of recombinant activated factor VII in trauma and surgery. AB - It is possible that recombinant activated factor VII (rFVIIa) could revolutionise the medical and surgical management of haemorrhage following trauma and surgery due to its ease of administration and mechanism of action. This article reviews the evidence for the use of rFVIIa as a pro-coagulant, its mechanism of action, safety and recent research into its use in blunt and penetrating trauma and haemorrhage. The potential role of rFVIIa, both in the pre-hospital environment and in the emergency room or operating theatre, is discussed. Administration of rFVIIa at the roadside, on the battlefield or in a trauma centre may have significant potential implications, especially for the paramedical and surgical teams and even for strategic planners. Finally, the areas where further research is needed to provide objective evidence of its efficacy are elaborated. The potential thromboembolic complications of systemic administration of rFVIIa need to be carefully monitored but the high cost of this drug is likely to be the limiting factor in its widespread use. PMID- 15116815 TI - Convalescence after colonic resection with fast-track versus conventional care. AB - BACKGROUND: Multi-modal rehabilitation programmes may improve early postoperative body composition, pulmonary function, exercise capacity, and reduce hospital stay. So far, no data are available on convalescence after discharge. AIM: The objectives were to compare convalescence data (fatigue, sleep, time to resume normal activities, and functional capabilities) and need for nursing care and contact to general practitioner with fast-track multi-modal rehabilitation compared with conventional care after colonic surgery. METHODS: Non-randomised, prospective controlled study in 30 consecutive patients undergoing fast-track rehabilitation with continuous epidural analgesia, enforced oral nutrition, mobilisation, planned early discharge, and 30 consecutive patients undergoing conventional care. Patients were interviewed preoperatively and 14 and 30 days postoperatively. RESULTS: Median hospital stay was 2 vs. 8 days in the fast-track vs. conventional care group, respectively (p < 0.01). Fourteen days postoperatively, total and mid-day sleep were increased in the conventional care group when compared with the fast-track group (p < 0.01). Fatigue was increased significantly at 14 days (p < 0.05) and throughout the study period compared with the fast-track group (p < 0.01). Similarly, ability to walking stairs, cooking, house keeping, shopping and walking outdoor was significantly less reduced at 14 days in the fast-track group, who also regained leisure activities earlier (p < 0.05). There was no significant difference between groups at 30 days or between need for nursing care and visits to general practitioners. Readmission for surgery-related events occurred more frequently (5 vs. 1 patient) in the fast track group. CONCLUSION: Fast-track rehabilitation with early discharge after colonic surgery results in earlier resumption of normal activities with reduced fatigue and need for sleep postoperatively compared to conventional care, and without increased need for nursing care or visits to general practitioners. However, readmissions may occur more frequently. PMID- 15116816 TI - Acute pancreatitis in Bergen, Norway. A study on incidence, etiology and severity. AB - BACKGROUND: Studies on the incidence and etiology of acute pancreatitis show large regional differences. This study was performed to establish incidence, etiology and severity of acute pancreatitis in the population of Bergen, Norway. METHODS: A study of all patients with acute pancreatitis admitted to Haukeland University Hospital over a 10-year period was performed. Information was obtained about the number of patients with acute pancreatitis admitted to the Deaconess Hospital in Bergen. RESULTS: A total of 978 admissions of acute pancreatitis were recorded in these two hospitals giving an incidence of 30.6 per 100,000. Haukeland University Hospital had 757 admissions of acute pancreatitis in 487 patients. Pancreatitis was severe in 20% (96/487) of patients, more often in males (25%) than in females (14%). Mortality due to acute pancreatitis was 3% (16/487). Gallstones were found to be an etiological factor in 48.5% and alcohol consumption in 19% of patients. The risk of recurrent pancreatitis was 47% in alcohol induced and 17% in gallstone induced pancreatitis. The last five years of the study period, endoscopic sphincterotomy of patients with gallstone pancreatitis, resulted in drop in relapse rate from 33% to 1.6%. CONCLUSION: The incidence of acute pancreatitis was found to be 30.6 per 100,000 with 48.5% associated with gallstones and 17% alcohol induced. Incidence of first attack was 20/100,000. Pancreatitis was classified as severe in 20% of cases with a mortality of 3%. PMID- 15116817 TI - Equal results with laparoscopic and Shouldice repairs of primary inguinal hernia in men. Report from a prospective randomised study. AB - AIM: To compare the laparoscopic and Shouldice techniques for repair of inguinal hernia. MATERIAL: 261 healthy men over 50 years with primary, unilateral inguinal hernia were randomly allocated to laparoscopic (total extraperitoneal approach, TEP) treatment (n = 131) or to a modified Shouldice technique (n = 130). RESULTS: Apart from a longer operative time in the laparoscopic group, there were no significant differences between the two methods with regard to perioperative complications, hospital stay, recurrencies or pain in the groin. CONCLUSIONS: Results following the total extraperitoneal laparoscopic and the Shouldice technique do not differ significantly 2 years after hernia repair. PMID- 15116819 TI - Primary hyperparathyroidism: intraoperative PTH-measurements. AB - BACKGROUND: With the development of rapid assays and intraoperative measurement of intact parathyroid hormone (PTH), new strategies in the handling of patients with primary hyperparathyroidism (pHPT) have evolved. AIM: The aim of our study was to illustrate the performance of the intraoperative PTH measurement as a predictor of successful cure. MATERIAL AND METHODS: From September 1999 to April 2002 143 patients with pHPT underwent a parathyroid operation (bilateral neck exploration with identification of all parathyroid glands) with intraoperative measurements of plasma PTH (immediately prior to surgery (T0) and 5 minutes after gland excision (T5)). A positive test result was defined as plasma PTH values at T5 below 20% of T0 or a value in the normal range below 7.6 pmol/l. Hence T5 values above 20% of T0 and above 7.6 pmol/l were considered test negative. RESULTS: 122 patients (85%) were test positive and cured, 11 patients (8%) were test negative but cured, and 10 patients (7%) were test negative and not cured by the primary operation. Consequently, the sensitivity of the test was 0.92 and the specificity 1.00. CONCLUSIONS: The rapid PTH test used is a reliable predictor of a successful outcome in pHPT patients undergoing parathyroid surgery. PMID- 15116818 TI - Outcome of surgery for oesophageal carcinoma in a low volume centre, with and without preoperative chemoradiotherapy. AB - AIMS: To critically assess the outcome of surgery for oesophageal carcinoma, with or without neoadjuvant chemoradiotherapy. METHODS: Since April 1998 until August 2002 resectable oesophageal cancer patients referred to us have received multimodal treatment, consisting of two courses of fluorouracil, cisplatin and hydroxyurea and 2 x 20 Gy of radiotherapy followed by surgery. The outcome of this treatment was compared to the outcome of a historical group of oesophageal cancer patients, treated with surgery alone in the time period 1994 to 1998. The patients represent a consecutive series of 20 resectable oesophageal carcinomas, referred to us since 1994. Four patients (20%) were treated for squamocellular carcinoma, 16 (80%) patients for adenocarcinoma. RESULTS: Treatment related toxicity was low and there was no death attributable to the chemoradiotherapy. Postoperative hospital mortality (< 30 days) and morbidity rates were 10% and 50%, respectively. A complete pathological response (T0) occurred in two of the nine patients in the multimodal group (22%). Overall median survival was 11 months. Median survival among patients in the multimodal group was 14 months, as compared with 7 months in the group treated with surgery alone (P = 0.041). CONCLUSIONS: Despite low volume, outcome of surgery for oesophageal carcinoma was acceptable. PMID- 15116820 TI - Dome down laparosonic cholecystectomy. AB - BACKGROUND AND AIMS: In cases of cholecystitis or other difficult situations conversion to laparotomy may be necessary. Furthermore, some cases of bile duct injury may be caused by anatomical variations and/or injuries caused by electrocautery. A new laparoscopic technique using a fundus-first technique with ultrasonic dissection ("dome down laparosonic cholecystectomy") may potentially solve these problems. MATERIAL AND METHODS: Twenty non-selected and consecutive patients with gallstone disease had laparoscopic cholecystectomy by the laparosonic dome down technique. Effect parameters were operative findings, duration of surgery and clinical course. RESULTS: Six patients had fibrotic chronic cholecystitis and one had acute cholecystitis. Three patients had intraoperative cholangiography because of elevated liver enzymes. All operations were completed by the intended laparosonic dome down technique. Median duration of surgery in the 20 patients was 35 minutes (range 20-60). Median duration of surgery in the first 5 patients was 55 minutes (35-60), and in the last 15 patients it was 30 minutes (20-50). All patients had uneventful clinical courses with a median hospital stay of 0 days (0-1). CONCLUSIONS: The technique was easy to learn and this new operative technique seems promising especially in cases of inflammation or fibrosis. PMID- 15116821 TI - Endovascular vs open AAA repair: similar effects on renal proximal tubular function. AB - AIM: To compare the effect of open and endovascular repair on renal function. MATERIALS AND METHODS: In a prospective, non-randomized study twenty-four abdominal aortic aneurysms (AAA) treatable with either method were repaired, 15 using endovascular device (ENDO group) and nine with open surgery with infrarenal aortic cross-clamping (OPEN group). All the patients had standardised general anaesthesia, intravascular fluid therapy and monitoring. Renal function tests and cardiovascular measurements were performed at predetermined intervals. RESULTS: N acetyl-beta-D-glucosaminidase indexed to urinary creatinine (U-NAG/crea), a sensitive marker of renal proximal tubular damage, increased similarly in both groups at the end of surgery (two-way ANOVA, p < 0.05). No patient developed clinical renal impairment, on the contrary, creatinine clearance was increased, serum cystatin C (a sensitive marker of renal glomerular filtration) and serum creatinine concentration decreased at 24 hours postoperatively (Wilcoxon paired test, p < 0.05). Intraoperative blood loss and the amount of administered crystalloids were higher in the OPEN than in the ENDO group (Mann-Whitney U-test, p < 0.05). The cardiovascular measurements were comparable between the groups. The mean (SD) amount of radio-contrast media given was 3.1 (1.1) ml/kg in the ENDO group. CONCLUSIONS: Our results indicate that endovascular AAA repair does not protect renal proximal tubular function. A temporary renal tubular dysfunction was found both in open and in endovascular AAA repair which did not lead to permanent changes in renal function. PMID- 15116822 TI - Surgical treatment of popliteal artery aneurysms. AB - OBJECTIVES: To report the results of surgical treatment of popliteal aneurysms with respect to symptoms and aneurysm size. DESIGN: A retrospective study based on prospectively registered data, in a single vascular unit. PATIENTS AND METHODS: Forty-nine patients were subjected to 57 operations from May 1974 to June 2000. Patency and limb salvage rates are compared for limbs with and without symptoms of ischaemia, and for small (2 cm or less) and large (> 2 cm) aneurysms. The long-term survival rate was calculated and compared with that of an age and sex-matched population. RESULTS: The overall 5 year graft patency was 60%. It was 83% for asymptomatic limbs and 49% for limbs with ischaemic symptoms. This difference was significant (p < 0.05). The overall 5 year limb salvage rate was 76%. It was 100% for asymptomatic and 64% for symptomatic limbs and this difference was significant (p < 0.05). Twenty-one of the aneurysms were 2 cm or less in diameter and 85% of these caused symptoms of ischemia. The operative mortality was 4%. The 5-year survival rate was 57% and significantly lower than that of a demographically matched population. CONCLUSION: The results of prophylactic operations for popliteal aneurysms on asymptomatic limbs are significantly better than those of operations done on limbs with ischaemic symptoms. The aneurysm size at which to recommend surgery is still not settled. PMID- 15116823 TI - Combined abdominal aortic aneurysmectomy and other abdominal operations. AB - BACKGROUND AND AIM: Co-existence of intra-abdominal non-vascular disease with an abdominal aortic aneurysm (AAA) poses a difficult surgical challenge. MATERIAL AND METHODS: Review of hospital records of 602 patients undergoing elective surgery for AAA during a 9-year period identified 61 (10.3%) patients with a co existent intra-abdominal non-vascular disease requiring surgery. RESULTS: The concomitant operations were 26 cholecystectomies, 11 inguinal hernia repairs, 2 small bowel resections, 5 left and 5 right hemicolectomies and 1 low anterior resection for colorectal carcinoma, 1 gastrectomy for gastric carcinoma, 5 nephrectomies, one salvage cystectomy for renal carcinoma and 1 left liver lobectomy for hepatrocellular carcinoma. Additional procedures for benign diseases prolonged the operative time by a mean of 35 (range 20-105) minutes and the major operations for malignancy by 120 (range 60-225) minutes. The overall hospital mortality and morbidity rates in the whole series of AAA (n = 602) remained as low as 0.66% and 13.6% respectively. There was no mortality and only two complications occurred in patients undergoing the combined procedure (n = 61). During a follow up period of 4-70 months, no graft infections were detected. CONCLUSION: In selected patients, the one stage approach is safe and effective. Attention should be given to the technical details and the rules of antisepsis. In elderly patients with AAA, a co-existent malignancy should be actively excluded. PMID- 15116824 TI - Cast vs external fixation: a comparative study in elderly osteoporotic distal radial fracture patients. AB - AIM: This study compared fracture treatment with plaster cast vs external fixation. METHODS: Forty elderly female osteoporotic wrist fracture patients were randomized to be treated with either plaster cast (Group A) or external fixation (Group B). Bone mineral density less than -2.5 T-score was among the inclusion criteria. RESULTS: In Group A, four redisplacements occurred, whereas in Group B there were none (p = 0.005). Horesh score was higher in Group B (p < 0.006) than in Group A. Volar angle deformity (p < 0.0005) and radial angle deformity (p = 0.008) were lower in Group B. CONCLUSIONS: This study shows that external fixation improves stability in elderly osteoporotic wrist fracture patients. PMID- 15116825 TI - The epidemiology and treatment patterns of postoperative adhesion induced intestinal obstruction in Varsinais-Suomi Hospital District. AB - BACKGROUND AND AIMS: The epidemiology and treatment patterns of postoperative adhesion induced intestinal obstruction have been poorly investigated in Finland. This study evaluated the epidemiology and treatment patterns of postoperative adhesion induced intestinal obstruction in a well defined geographical area (Hospital District). MATERIAL AND METHODS: All inpatient episodes between 1.1.1999 and 31.12.1999 due to postoperative adhesion induced intestinal obstruction in Varsinais-Suomi Hospital District were evaluated retrospectively using individual patient records. RESULTS: 123 hospitalizations due to postoperative adhesion-related intestinal obstruction were observed during the study period. The total number of preceding operations was 176 considering altogether 101 patients. The most prevalent single initial operations causing adhesion induced intestinal obstruction were colorectal, upper abdominal, and female reproductive system procedures. Of all treatment episodes 32% were operative and mortality was 2%. The median days of hospital stay (range) of all inpatient episodes, operative episodes, and conservative episodes were 6 (1-58), 11 (2-34) and 4 (1-58), respectively. Patient dependent factors associated with increased likelihood to operative treatment of obstruction were: female gender (40% in females vs 23% in males, P = 0.042) and previous gynaecological surgery (70% of the patients, P = 0.032). Intraoperative findings were obstruction in 70%, strangulation in 20%, necrosis in 8%, and perforation in 2% of operations. Bowel resection was needed in 38% of operations. Preceding gynaecological surgery increased the likelihood of bowel strangulation as an intraoperative finding. CONCLUSION: The epidemiology, treatment patterns and results of postoperative adhesion induced intestinal obstruction are of the average international level in the Varsinais-Suomi Hospital District. The treatment patterns among the different hospitals in the Hospital District are similar. Female gender is associated with increased risk for operative treatment of adhesive obstruction. Previous gynaecological surgery increases the likelihood of operative treatment and complicated obstruction. PMID- 15116826 TI - Glove perforations in open and laparoscopic abdominal surgery: the feasibility of double gloving. AB - BACKGROUND AND AIMS: According to the traditional view, the glove protects the patient from the bacterial growth of the surgeons' hands and doing so prevents infections. Today, with growing incidences of HIV and Hepatitis B and C, surgical gloves are also important as protection for the surgeon. We compared the safety of double indicator gloves to standard single surgical gloves by investigating how often surgical gloves are punctured in laparoscopic and open gastrointestinal surgery. STUDY: As study material we gathered all gloves that had been used in gastrointestinal surgery in Satakunta Central Hospital during two months. 814 gloves from 274 operations were tested by using standardized water filling test method. RESULTS: In open surgery 67 gloves out of 694 had been punctured (9.6 percent). Puncture occurred in 22.5 percent of operations (53 out of 236). During open surgery 24 holes out of 35 were undetected with single gloves (69 percent). With double indicator gloves, only 3 out of 31 holes were unnoticed (10 percent). Long duration of operation increased the risk of puncture. In laparoscopic operations 4 gloves out of 120 had been perforated (3.3 percent). CONCLUSION: Double surgical gloves give markedly better protection in surgery. This is important especially in high risk operations. PMID- 15116828 TI - Cooking spews out ultrafine particles. PMID- 15116827 TI - Morgagni hernia in adults: results in 7 patients. AB - OBJECTIVE: Morgagni's hernia is a relatively uncommon diaphragmatic hernia with a potential for considerable morbidity, if the diagnosis is delayed or missed. This review of cases of Morgagni's hernia was undertaken in order to emphasize methods of diagnosis and treatment. METHODS: From 1992 through 2002, seven patients with Morgagni's hernia (5 right, 2 left) were surgically treated at our hospital. We investigated the patients preoperatively including chest roentgenogram, chest CT scan, and contrast studies of the upper gastrointestinal tract. Operative repair was accomplished with the transabdominal or transthoracic approach. Basic spirometric tests had been carried out on patients presented for elective surgery. RESULTS: The majority of patients experienced dyspnea and two patients presented with acute abdomen due to peritonitis. Diagnosis for Morgagni's hernia was made preoperatively in all but one patient. In cases with uncertain diagnosis or peritonitis, a transabdominal approach was preferred. One patient had died of septic multi-organ failure in the early postoperative course. Following elective repair of Morgagni's hernia, improvement in basic spirometric values was seen. CONCLUSIONS: We conclude that repair for Morgagni's hernia can be performed safely and effectively by using different surgical approaches. The risk of progression and incarceration makes clinical awareness, early diagnosis, and surgical treatment warranted. Improvement in lung function can be expected postoperatively. PMID- 15116829 TI - White House denies scientific manipulation. PMID- 15116830 TI - Administration drops pesticide consultations. PMID- 15116831 TI - Mercury woes appear to grow. PMID- 15116832 TI - Direct identification of trace metals in fine and ultrafine particles in the Detroit urban atmosphere. AB - Exposure to airborne particulates containing low concentrations of heavy metals, such as Pb, As, and Se, may have serious health effects. However, little is known about the speciation and particle size of these airborne metals. Fine- and ultrafine particles with heavy metals in aerosol samples from the Detroit urban area, Michigan, were examined in detail to investigate metal concentrations and speciation. The characterization of individual particles was completed using high angle annular dark-field scanning transmission electron microscopy (HAADF-STEM) combined with conventional high-resolution TEM techniques. The trace elements, Pb, As, La, Ce, Sr, Zn, Cr, Se, Sn, Y, Zr, Au, and Ag, were detected, and the elemental distributions were mapped in situ atthe nanoscale. The crystal structures of the particles containing Pb, Sr, Zn, and Au were determined from their electron diffraction patterns. Based on the characterization of the representative trace element particles, the potential health effects are discussed. Most of the trace element particles detected in this study were within a range of 0.01-1.0 microm in size, which has the longest atmospheric residence time (approximately 100 days). Increased chemical reactivity owing to the size of nanoparticles may be expected for most of the trace metal particles observed. PMID- 15116833 TI - Biomagnification of alpha- and gamma-hexabromocyclododecane isomers in a Lake Ontario food web. AB - The extent of bioaccumulation of hexabromocyclododecane (HBCD) isomers (alpha, beta, and gamma) was determined in the Lake Ontario pelagic food web using liquid chromatography tandem mass spectrometry (LC/MS/MS). Concentrations of the alpha isomer were consistently higher than that of the gamma-isomer. The beta-isomer was below method detection limits in all samples. Whole body concentrations (ng/g, wet wt) of alpha- and gamma-HBCD were highest in the top predator lake trout samples ranging from 0.4 to 3.8 ng/g for the alpha-isomer and 0.1 to 0.8 ng/g for the gamma-isomer. For the prey fish species, the trends in alpha- and gamma-HBCD levels were slimy sculpin > smelt > alewife. Mean concentrations of total (sigma) HBCD (sum of alpha- and gamma-isomers) in the macrozooplankter Mysis relicta (0.14 +/- 0.02 ng/g wet wt) and in the benthic invertebrate Diporeia hoyi (0.16 +/- 0.02 ng/g, wet wt) were similar and approximately twice as high as in plankton (0.06 +/- 0.02 ng/g, wet wt). A strong positive linear relationship was found between sigmaHBCD concentrations (wet wt) and trophic level based on delta15N suggesting that HBCD biomagnifies in the Lake Ontario food web. The trophic magnification factor (TMF = 6.3) derived from the slope of the sigmaHBCD - trophic level relationship was slightly higher than TMFs for p,p' DDE (6.1) and sigmaPCBs (5.7) found previously. Biomagnification factors (BMF, calculated as the ratio of lipid corrected concentration in predator/lipid corrected concentration in prey) were variable between feeding relationships and ranged from 0.4 to 10.8 for the alpha-isomer and from 0.2 to 10 for gamma isomers. PMID- 15116834 TI - Source strengths of ultrafine and fine particles due to cooking with a gas stove. AB - Cooking, particularly frying, is an important source of particles indoors. Few studies have measured a full range of particle sizes, including ultrafine particles, produced during cooking. In this study, semicontinuous instruments with fine size discriminating ability were used to calculate particle counts in 124 size bins from 0.01 to 2.5 microm. Data were collected at 5 min intervals for 18 months in an occupied house. Tracer gas measurements were made every 10 min in each of 10 rooms of the house to establish air change rates. Cooking episodes (N = 44) were selected meeting certain criteria (high concentrations, no concurrent indoor sources, long smooth decay curves), and the number and volume of particles produced were determined for each size category. For each episode, the particle decay rate was determined and used to determine the source strength for each size category. The selected cooking episodes (mostly frying) were capable of producing about 10(14) particles over the length of the cooking period (about 15 min), more than 90% of them in the ultrafine (< 0.1 microm) range, with an estimated whole house volume concentration of 50 (microm/cm)3. More than 60% of this volume occurred in the 0.1-0.3 microm range. Frying produced peak numbers of particles at about 0.06 microm, with a secondary peak at 0.01 microm. The peak volume occurred at a diameter of about 0.16 microm. Since the cooking episodes selected were biased toward higher concentrations, the particle concentrations measured during about 600 h of morning and evening cooking over a full year were compared to concentrations measured during noncooking periods at the same times. Cooking was capable of producing more than 10 times the ultrafine particle number observed during noncooking periods. Levels of PM2.5 were increased during cooking by a factor of 3. Breakfast cooking (mainly heating water for coffee and using an electric toaster) produced concentrations about half those produced from more complex dinnertime cooking. Although the number and volume concentrations observed depend on air change rates, house volume, and deposition rates due to fans and filters, the source strengths calculated here are independent of these variables and may be used to estimate number and volume concentrations in other types of homes with widely varying volumes, ventilation rates, and heating and air-conditioning practices. PMID- 15116835 TI - Global pollution monitoring of polybrominated diphenyl ethers using skipjack tuna as a bioindicator. AB - To elucidate the global distribution of polybrominated diphenyl ethers (PBDEs), these chemicals were determined in the muscle of skipjack tuna (Katsuwonus pelamis) collected from offshore waters of various regions in the world (Japan, Taiwan, Philippines, Indonesia, Seychelles, and Brazil, and the Japan Sea, East China Sea, South China Sea, Indian Ocean, and North Pacific Ocean). PBDEs were detected in almost all the skipjack tuna collected from the locations surveyed (from < 0.1 to 53 ng/g of lipid), indicating widespread contamination by these compounds in the marine environment. Residue levels of PBDEs in these samples from the northern hemisphere seem to be higher than those from the southern hemisphere, which is plausibly due to larger usage of these compounds in the northern hemisphere. Higher concentrations of PBDEs were detected in the samples from waters around the East China Sea (up to 53 ng/g of lipid). Developing countries around the East China Sea are supposedly the "hot spots" releasing these chemicals into the marine environment. With regard to the composition of PBDE congeners, the percentage contribution by lower brominated congeners (BDE15, -28, and -47) showed an increasing trend with increasing latitude. On the other hand, higher brominated congeners (BDE153, -154, and -183) showed a reverse trend. These patterns suggest that lower brominated congeners of PBDEs (di-, tri , and tetra-BDEs) were preferentially transported from pollution sources to northern colder regions through the atmosphere. PBDEs may have a high potency to cause global pollution like PCBs. PMID- 15116836 TI - Receptor model comparisons and wind direction analyses of volatile organic compounds and submicrometer particles in an arid, binational, urban air shed. AB - The relationship between continuous measurements of volatile organic compounds sources and particle number was evaluated at a Photochemical Assessment Monitoring Station Network (PAMS) site located near the U.S.-Mexico Border in central El Paso, TX. Sources of volatile organic compounds (VOCs) were investigated using the multivariate receptor model UNMIX and the effective variance least squares receptor model known as Chemical Mass Balance (CMB, Version 8.0). As expected from PAMS measurements, overall findings from data screening as well as both receptor models confirmed that mobile sources were the major source of VOCs. Comparison of hourly source contribution estimates (SCEs) from the two receptor models revealed significant differences in motor vehicle exhaust and evaporative gasoline contributions. However, the motor vehicle exhaust contributions were highly correlated with each other. Motor vehicle exhaust was also correlated with the ultrafine and accumulation mode particle count, which suggests that motor vehicle exhaust is a source of these particles at the measurement site. Wind sector analyses were performed using the SCE and pollutant data to assess source location of VOCs, particle count, and criteria pollutants. Results from this study have application to source apportionment studies and mobile source emission control strategies that are ongoing in this air shed. PMID- 15116837 TI - Long-term recovery of PCB-contaminated surface sediments at the Sangamo-westonl Twelvemile Creek/lake Hartwell Superfund Site. AB - Natural recovery of contaminated sediments relies on burial of contaminated sediments with increasingly clean sediments over time (i.e., natural capping). Natural capping reduces the risk of resuspension of contaminated surface sediments, and it reduces the potential for contaminant transport into the food chain by limiting bioturbation of contaminated surface or near-surface sediments. This study evaluated the natural recovery of surface sediments contaminated with polychlorinated biphenyls (PCBs) at the Sangamo-Weston/Twelvemile Creek/Lake Hartwell Superfund Site (Lake Hartwell), Pickens County, SC. The primary focus was on sediment recovery resulting from natural capping processes. Total PCB (t PCB), lead-210 (210Pb), and cesium-137 (137Cs) sediment core profiles were used to establish vertical t-PCB concentration profiles, age date sediments, and determine surface sedimentation and surface sediment recovery rates in 18 cores collected along 10 transects. Four upgradient transects in the headwaters of Lake Hartwell were impacted by historical sediment releases from three upgradient sediment impoundments. These transects were characterized by silt/ clay and sand layering. The highest PCB concentrations were associated with silt/clay layers (1.8-3.5% total organic carbon (TOC)), while sand layers (0.05-0.32% TOC) contained much lower PCB concentrations. The historical sediment releases resulted in substantial burial of PCB-contaminated sediment in the vicinity of these four cores; each core contained less than 1 mg/kg t-PCBs in the surface sand layers. Cores collected from six downgradient Lake Hartwell transects consisted primarily of silt and clay (0.91-5.1% TOC) and were less noticeably impacted by the release of sand from the impoundments. Vertical t-PCB concentration profiles in these cores began with relatively low PCB concentrations at the sediment-water interface and increased in concentration with depth until maximum PCB concentrations were measured at approximately 30-60 cm below the sediment-water interface, ca. 1960-1980. Maximum t-PCB concentrations were followed by progressively decreasing concentrations with depth until the t-PCB concentrations approached the detection limit, where sediments were likely deposited before the onset of PCB use at the Sangamo-Weston plant. The sediments containing the maximum PCB concentrations are associated with the period of maximum PCB release into the watershed. Sedimentation rates averaged 2.1 +/- 1.5 g/(cm2 yr) for 12 of 18 cores collected. The 1994 Record of Decision cleanup requirement is 1.0 mg/kg; two more goals (0.4 and 0.05 mg/kg t PCBs) also were identified. Average surface sedimentation requirements to meet the three goals were 1.4 +/- 3.7, 11 +/- 4.2, and 33 +/- 11 cm, respectively. Using the age dating results, the average recovery dates to meet these goals were 2000.6 +/- 2.7, 2007.4 +/- 3.5, and 2022.7 +/- 11 yr, respectively. (The 95% prediction limits for these values also are provided.) Despite the reduction in surface sediment PCB concentrations, PCB concentrations measured in largemouth bass and hybrid bass filets continue to exceed the 2.0 mg/kg FDA fish tolerance level. PMID- 15116839 TI - Preliminary assessment of U.K. human dietary and inhalation exposure to polybrominated diphenyl ethers. AB - This study reports concentrations of BDEs 47, 99, 100, 153, and 154 in outdoor air [median sigmaPBDE (sum of BDEs 47, 99, 100, 153, and 154) = 18 pg m(-3)] in air from a range of office and home indoor microenvironments (median sigmaPBDE = 762 pg m(-3)) and vegan and omnivorous duplicate diet samples (median sigmaPBDE = 154 and 181 pg g(-1) dryweightforvegan and omnivorous diets, respectively). Median daily human exposure to sigmaPBDE via inhalation is 6.9 ng/person and 90.5 ng/person via diet but the relative significance of these pathways may vary considerably between individuals. Median concentrations in indoor air were higher in workplace (sigmaPBDE = 1082 pg m(-3)) than in domestic (sigmaPBDE = 128 pg m( 3)) microenvironments, and substantial differences in concentrations in air from different rooms in the same office building were found. When data from the only mechanically ventilated room was excluded, a significant positive correlation (p < 0.001) was observed between PBDE concentrations and both the number of electrical appliances and polyurethane foam-containing chairs. Concentrations of sigmaPBDE and BDEs 47 and 99 were significantly higher (p < 0.1) in omnivorous diet samples than in vegan diet samples, implying that while plant-based foods contribute appreciably, higher exposure occurs via ingestion of animal-based comestibles. PMID- 15116838 TI - Polybromodiphenyl ether flame retardants in fish from lakes in European high mountains and Greenland. AB - Individual polybromodiphenyl ethers (PBDEs) were investigated in liver and muscle tissue of trout from 11 high mountain lakes in Europe and one in Greenland. Trouts in these lakes [brown trout (Salmo trutta), brook trout (Salvelinus fontinalis) and arctic char (Salvelinus alpinus)] are important sentinel species because they are located in the top of the food chain and pollution can only reach these ecosystems by atmospheric transport. The major PBDE congeners were BDE 47 and BDE 99, followed by BDE 100, BDE 153, BDE 154, and BDE 28. These compounds were found in all the samples examined. Their average concentrations [110-1300 and 69-730 pg g(-1) wet weight (ww) in liver and muscle or 2400-40000 and 2900-41000 pg g(-1) lipid weight (lw), respectively] were in the lower range when compared with those of fish from other less remote locations. The highest levels of PBDEs in liver and muscle are found in Lochnagar, Scotland: 11000 and 1200 pg g(-1) ww, respectively (366 000 and 177000 pg g(-1) lw, respectively). Male specimens exhibited higher PBDE concentrations in liver than female. The concentrations of most PBDEs in liver were correlated with fish age (p < 0.01) and, inversely, with condition factor (p < 0.01). Muscle PBDE concentrations did not correlate with age, and only some congeners showed significant positive correlations with condition factor (p < 0.05). The main differences between species were found in the accumulation of the more abundant PBDEs, brook trout showing the highest concentrations in muscle and the lowest in liver. No correlation between the occurrence of these compounds in high mountain fish and altitude, latitude, or temperature was observed. This fact and the lack of correlation between muscle concentrations and age suggest that the fluxes of PBDEs arriving at high mountain lakes are still not constant. In view of the present use of these compounds, they are probably increasing. PMID- 15116840 TI - Effects of dissolved organic matter surrogates on the partitioning of 17beta estradiol and p-nonylphenol between synthetic membrane vesicles and water. AB - Partition coefficients of two estrogenic compounds, 17beta-estradiol (E2) and p nonylphenol (NP), between synthetic membrane vesicles (K'lipw values) and water were determined using equilibrium dialysis to evaluate the potential biological uptake of these compounds with and without the presence of dissolved natural organic matter (DOM) surrogates, Suwannee River fulvic acid, dialyzed Aldrich humic acid, and polyphenolic tannic acid. Overall, K'lipw values for E2 and NP reduced by 20-30% with the increase of DOM concentration from 0 to approximately 4 mg of C/L, a typical DOM concentration in the aquatic environment. This trend for E2 and NP is similar to that obtained by other researchers for moderately hydrophobic compounds including polycyclic aromatic hydrocarbons with three or four rings. A three-phase compartment model with three independent interactions, the sorption of E2 or NP by DOM surrogate (Koc), the sorption of DOM surrogates by membrane vesicles (KlipDOM), and the partition of E2 or NP by membrane vesicles (Klipw), was proposed, and K'lipw values for E2 or NP in the presence of DOM were calculated. The model predicted the decrease in Klipw values with the increase of DOM concentration, and the predictions using the three linear interactions agreed satisfactorily with the experimental results at relatively lower concentration of DOM. PMID- 15116841 TI - Biodegradation of linear alkylbenzene sulfonates and their degradation intermediates in seawater. AB - A study has been made of the aerobic biodegradation of linear alkylbenzene sulfonates (C12 and C11 homologues) and sulfophenylcarboxylic acids (C5 and C11 homologues) in seawater at concentrations of the same order as those detected ones in coastal waters influenced by wastewater effluents, at different temperatures, and both with and without the addition of an inoculum adapted to the presence of linear alkylbenzene sulfonate (LAS). The biodegradation of C12LAS, C11LAS, C5SPC, and C11SPC exceeds 99% in all tests performed and can be satisfactorily fitted to a second-degree polynomial without an independent term. The kinetic of degradation of LAS presents a clear seasonal component, since the process is considerably inhibited at lower temperatures; it is also kinetically enhanced by the presence of the inoculum. The intermediates detected for all the cases are sulfophenylcarboxylic acids (SPCs), the most abundant being those intermediates produced by the omega- and beta-oxidations of the parent compound, although intermediates produced by the alpha-oxidation have also been detected. The kinetic of the SPCs generated can be described using a model composed of two terms that represent the formation and the degradation of these intermediates. The total disappearance of the SPCs in all cases indicates that the degradation of LAS in seawater at the tested concentrations in aerobic conditions is complete. PMID- 15116842 TI - Arsenic removal from aqueous solution via ferrihydrite crystallization control. AB - Removal of arsenate anion from aqueous solution by coprecipitation with ferrihydrite has been studied under conditions in which the Fe/As ratio is maintained at a constant level, while the degree of supersaturation with respect to the iron oxide precipitate is varied. An Fe/As ratio of 12 was chosen, and supersaturation was controlled by varying the iron concentration or the pH. The relationship between supersaturation and arsenic removal was found to follow an exponential curve, with greater arsenic removal occurring at higher supersaturation ratios for each of the pH values tested. Higher supersaturation ratios were required to achieve a given level of arsenic removal at pH 7 than would be required to achieve the same level of removal at pH 3.5. The results provide important guidelines for selection of appropriate concentrations of iron(III) required for arsenic removal under various circumstances. Powder XRD analysis of the arsenate-ferrihydrite precipitates showed an increasing degree of structural order with decreasing levels of supersaturation. TEM images of the precipitates revealed that aggregates with a morphology similar to that of schwertmannite are formed in some samples at low supersaturation levels. The results described in this paper indicate that the overall efficiency of arsenic removal involves a combination of both supersaturation and pH effects, with pH controlling the affinity of arsenate for the ferrihydrite surface, and supersaturation controlling the surface area and physical properties of the ferrihydrite product. PMID- 15116843 TI - Fate of elemental mercury in the Arctic during atmospheric mercury depletion episodes and the load of atmospheric mercury to the Arctic. AB - Atmospheric mercury depletion episodes (AMDEs) were studied at Station Nord, Northeast Greenland, 81 degrees 36' N, 16 degrees 40' W, during the Arctic Spring. Gaseous elemental mercury (GEM) and ozone were measured starting from 1998 and 1999, respectively, until August 2002. GEM was measured with a TEKRAN 2735A automatic mercury analyzer based on preconcentration of mercury on a gold trap followed by detection using fluorescence spectroscopy. Ozone was measured by UV absorption. A scatter plot of GEM and ozone concentrations confirmed that also at Station Nord GEM and ozone are linearly correlated during AMDEs. The relationship between ozone and GEM is further investigated in this paper using basic reaction kinetics (i.e., Cl, ClO, Br, and BrO have been suggested as reactants for GEM). The analyses in this paper show that GEM in the Arctic troposphere most probably reacts with Br. On the basis of the experimental results of this paper and results from the literature, a simple parametrization for AMDE was included into the Danish Eulerian Hemispheric Model (DEHM). In the model, GEM is converted linearly to reactive gaseous mercury (RGM) over sea ice with temperature below -4 degrees C with a lifetime of 3 or 10 h. The new AMDE parametrization was used together with the general parametrization of mercury chemistry [Petersen, G.; Munthe, J.; Pleijel, K.; Bloxam, R.; Vinod Kumar, A. Atmos. Environ. 1998, 32, 829-843]. The obtained model results were compared with measurements of GEM at Station Nord. There was good agreement between the start and general features periods with AMDEs, although the model could not reproduce the fast concentration changes, and the correlation between modeled and measured values decreased from 2000 to 2001 and further in 2002. The modeled RGM concentrations over the Arctic in 2000 were found to agree well with the temporal and geographical variability of the boundary column of monthly average BrO observed by the GOME satellite. Scenario calculations were performed with and without AMDEs. For the area north of the Polar Circle, the mercury deposition increases from 89 tons/year for calculations without an AMDE to 208 tons/year with the AMDE. The 208 tons/year represent an upper limit for the mercury load to the Artic. PMID- 15116844 TI - Climate effects on stream nitrate concentrations at 16 forested catchments in south central Ontario. AB - Increased nitrate (NO3) concentrations in streamwaters draining forested catchments are reportedly an early indicator of nitrogen (N) saturation. Nitrate concentrations in streams draining 16 forested catchments in south central Ontario were monitored over a 16-year period, during which time N bulk deposition was relatively constant (approximately 9 kg ha(-1) yr(-1)). Mean annual NO3 concentrations in streams were both highly variable among catchments and among years, although patterns of annual concentration were similar among many catchments. Coherence analysis identified two stream groupings. Shallow soils, moderate slopes, low NO3 concentration, and a large wetland component characterized the first group. The second group had primarily upland characteristics including deeper soils, steeper slopes, higher NO3 concentrations, and a much smaller wetland component. Patterns in NO3 concentration in wetland-influenced streams appeared to be related to summer drought and cumulative frost depth, whereas NO3 concentrations in upland-draining streams appeared to be related to both mean annual air temperature and summer drought. Because a number of different climate parameters as well as the physical character of the catchments apparently influence NO3 export, NO3 concentrations in streams are not a good indicator of N saturation in this region. PMID- 15116845 TI - Environmental fate of bisphenol A and its biological metabolites in river water and their xeno-estrogenic activity. AB - Monitoring of bisphenol A [BPA; 2,2-bis(4-hydroxyphenyl)propane] and its biological metabolites [4,4'-dihydroxy-alphamethylstilbene (DHMS), 2,2-bis(4 hydroxyphenyl)-1-propanol (BPA-OH), 2,2-bis(4-hydroxyphenyl)propanoic acid (BPA COOH), and 2-(3,4-dihydroxyphenyl)-2-(4-hydroxyphenyl)propane (3-OH-BPA)] in river waters was performed by solid-phase extraction and GC/MS determination. The concentrations of BPA, BPA-COOH, BPA-OH, and 3-OH-BPA in the river water ranged from 2 to 230 (8.8 x 10(-12) to 1.0 x 10(-9) M), from 5 to 75 (1.9 x 10(-11) to 2.9 x 10(-10) M), from 3 to 16 (1.2 x 10(-11) to 6.6 x 10(-11) M), and from 3 to 11 (1.2 x 10(-11) to 4.5 x 10(-11) M) ng L(-1), respectively. DHMS, an intermediate in the main degradation pathway of BPA, was not detected in any water sample. Under the aerobic conditions in the river water, BPA disappeared within 8 d of incubation, but BPA-COOH, BPA-OH, and tetraol remained in the supernatant after 14 d of incubation. For the xeno-estrogenic activity of BPA and the metabolites, their ability to bind to recombinant human estrogen receptor alpha in competition with fluorescence-labeled 17beta-estradiol was measured. Fifty percent inhibitory concentrations (IC50) of BPA, DHMS, 3-OH-BPA, and BPA-OH were approximately 1 x 10(-5), 1 x 10(-6), 3 x 10(-5), and 1 x 10(-2) M, respectively. In human cultured MCF-7 breast cancer cells, BPA increased cell numbers in a dose-dependent manner at concentrations from 10(-7) to 10(-5) M. For the BPA metabolites, DOHMS, 3-OH-BPA, and BPA-COOH caused the cells proliferation at concentrations from 10(-9) to 10(-6), from 10(-7) to 10(-6), and from 10(-5) to 10(-4) M, respectively. BPA-OH did not cause MCF-7 cells proliferation. These results indicate that BPA is mainly metabolized through oxidative rearrangement by bacteria in the river water, and intermediate bisphenols via minor metabolic pathways exist in river water. The presence of the bisphenols having the xeno estrogenic effect suggests the necessity of monitoring those in river water, in the effluent waters from sewage plants, or in landfill leachate. PMID- 15116846 TI - Metal speciation dynamics and bioavailability: bulk depletion effects. AB - Under conditions of bulk depletion, the speciation and bioavailability of trace metals must be considered at two different time scales: (i) the time scale of the biouptake flux, as determined by diffusion of the bioactive free metal, dissociation of the bioinactive complex species, and the internalization rate; and (ii) the time scale of depletion of the bulk medium. The implications of these two time scales for the speciation dynamics are discussed in terms of experimental conditions. The geometry of the system is taken into accountvia a spherical cellular model. It considers a spherical organism depleting a spherical volume in a nonstirred medium and assumes linear adsorption of the metal atthe biointerface and first-order internalization kinetics. In cases where the rate of biouptake is fully controlled by the internalization step, concentration gradients in the medium are insignificant. Then the biouptake becomes independent of the geometry of the system, and the model has a much simpler solution. Examples of trace metal uptake by microorganisms are analyzed: (i) cobalt uptake by Prochlorococcus in the presence of NTA, under conditions where bulk depletion is the controlling process due to the large number of organisms and high internalization rates, (ii) silver uptake by Chlamydomonas reinhardtii with significant effects of bulk depletion, due to the high internalization rate; (iii) lead uptake by Chlorella vulgaris with pratically negligible bulk depletion due to the low internalization rate of the metal; and (iv) lead uptake by intestinal Caco-2 cells, illustrating the simplification of the bulk depletion model for a system with different geometry where internalization is the rate controlling step. PMID- 15116847 TI - Bioaccumulation potential of persistent organic chemicals in humans. AB - A model was used to explore the influence of physicalchemical properties on the potential of organic chemicals to bioaccumulate in humans. ACC-HUMAN, a model of organic chemical bioaccumulation through the agricultural and aquatic food chains to humans, was linked to a level I unit world model of chemical fate in the physical environment and parametrized for conditions in southern Sweden. Hypothetical, fully persistent chemicals with varying physical-chemical properties were distributed in the environment, and their bioaccumulation to humans was calculated. The results were evaluated using the environmental bioaccumulation potential (EBAP), defined as the quotient of the chemical quantity in a human divided by the quantity of chemical in the whole environment. Since the latter is closely related to emissions, EBAP is potentially a more useful tool for comparative risk assessment of chemicals than currently used medium-specific measures such as the fish-water bioaccumulation factor. A high environmental bioaccumulation potential, defined as > 10% of the maximum EBAP, was found for chemicals with 2 < log KOW < 11 and 6 < log KOA < 12. While these chemical partitioning properties clearly influenced bioaccumulation at each trophic level, these effects tended to equalize over the food web. The fact that the transfer from the environment as a whole to humans was quite uniform over a large chemical partitioning space suggests that these partitioning properties are relatively unimportant determinants of human exposure compared to other factors such as the substance's persistence in the environment and in the food web. PMID- 15116848 TI - Uncertainty in particle number modal analysis during transient operation of compressed natural gas, diesel, and trap-equipped diesel transit buses. AB - The relationships between transient vehicle operation and ultrafine particle emissions are not well-known, especially for low-emission alternative bus technologies such as compressed natural gas (CNG) and diesel buses equipped with particulate filters/traps (TRAP). In this study, real-time particle number concentrations measured on a nominal 5 s average basis using an electrical low pressure impactor (ELPI) for these two bus technologies are compared to that of a baseline catalyst-equipped diesel bus operated on ultralow sulfur fuel (BASE) using dynamometer testing. Particle emissions were consistently 2 orders of magnitude lower for the CNG and TRAP compared to BASE on all driving cycles. Time resolved total particle numbers were examined in terms of sampling factors identified as affecting the ability of ELPI to quantify the particulate matter number emissions for low-emitting vehicles such as CNG and TRAP as a function of vehicle driving mode. Key factors were instrument sensitivity and dilution ratio, alignment of particle and vehicle operating data, sampling train background particles, and cycle-to-cycle variability due to vehicle, engine, after treatment, or driver behavior. In-cycle variability on the central business district (CBD) cycle was highest for the TRAP configuration, but this could not be attributed to the ELPI sensitivity issues observed for TRAP-IDLE measurements. Elevated TRAP emissions coincided with low exhaust temperature, suggesting on road real-world particulate filter performance can be evaluated by monitoring exhaust temperature. Nonunique particle emission maps indicate that measures other than vehicle speed and acceleration are necessary to model disaggregated real-time particle emissions. Further testing on a wide variety of test cycles is needed to evaluate the relative importance of the time history of vehicle operation and the hysteresis of the sampling train/dilution tunnel on ultrafine particle emissions. Future studies should monitor particle emissions with high resolution real-time instruments and account for the operating regime of the vehicle using time-series analysis to develop predictive number emissions models. PMID- 15116849 TI - Measurement of contemporary and fossil carbon contents of PM2.5 aerosols: results from Turtleback Dome, Yosemite National Park. AB - The impact of aerosol particulate matter of mean mass aerodynamic diameter < or = 2.5 microm (PM2.5 aerosols) on health, visibility, and compliance with the U.S. EPA's regional haze regulations is a growing concern. Techniques that can help better characterize particulate matter are required to better understand the constituents, causes, and sources of PM2.5 aerosols. Measurement of the 14C/C ratio of the PM2.5 aerosols, the absence of 14C in fossil carbon materials, and the known 14C/C levels in contemporary carbon materials allow the use of a two component model to derive contemporary and fossil carbon contents of the particulate matter. Such data can be used to estimate the relative contributions of fossil fuels and biogenic aerosols to the total aerosol loading. Here, the methodology for performing such an assessment using total suspended particulate hi-vol aerosol samplers to collect PM2.5 aerosols on quartz fiber filters and the technique of accelerator mass spectrometry to measure 14C/C ratios is presented and illustrated using PM2.5 aerosols collected at Yosemite National Park. PMID- 15116850 TI - Adsorption of arsenic from water using activated neutralized red mud. AB - In this paper activated seawater-neutralized red mud, herein referred to as activated Bauxsol (AB), is used as a novel adsorbent for removing inorganic arsenic (As) from water. The adsorption of As onto AB is studied as a function of contact time, particle size, pH, initial As concentration, AB dosage, and temperature. Kinetic data indicate that the process pseudoequilibrates in 3 and 6 h for As(V) (arsenate) and As(III) (arsenite), respectively, and follows a pseudo first-order rate expression. Within the range tested, the optimal pH for As(V) adsorption is 4.5, and close to 100% removal can be achieved irrespective of the initial As(V) concentration. Desorption of As(V) is greatest at pH 11.6 where a maximum of 40% can be achieved. In contrast, the optimum pH for As(III) removal is 8.5, and the removal efficiency changes with the initial As(III) concentration. The adsorption data fit the Langmuir isotherm and its linearized form well, with thermodynamic data indicating the spontaneous and endothermic nature of the process. The FITEQL (V.4) and PHREEQC (V.2) computer programs are used to predict As(V) adsorption at various pH values (based on diffuse double layer models). The modeling results fit the experimental results very well and indicate that surface complexation modeling is useful in describing the complex AB surface during the adsorption process. This study shows that As(III) needs to be oxidized to As(V) for a favorable removal using AB and that AB can be a very efficient unconventional adsorbent for removing As(V) from water. PMID- 15116851 TI - Removal of ionic dyes from water by solvent extraction using reverse micelles. AB - Several methods (e.g., UV/H2O2 oxidation, adsorption, flocculation-precipitation) are normally employed to remove dye from water. A new technique based on liquid/liquid extraction using reverse micelles is proposed whereby recovery of solvent and reuse of dye is possible. Experiments were conducted by mixing a known quantity of dye in aqueous phase and solvent-containing surfactants in a simple mixer. The separation of solvent phase, containing encapsulated dye in reverse micelles, from aqueous phase due to gravity results in separation of dye from water. The removal of different ionic dyes (e.g., eosin yellow, methylene blue, malachite green, methyl orange, orange G) from aqueous phase in the presence of different cationic and anionic surfactants [e.g., sodium dodecylbenzene sulfonate, sodium bis(2-ethylhexyl) sulfosuccinate, hexadecyltrimethylammonium bromide, and cetyl pyridinium chloride] in different solvents (e.g., amyl alcohol, benzyl alcohol, methyl benzoate, and isooctane) were studied by conducting experiments. The percentage removal of dye from aqueous phase increases with the decrease in dye concentration or with the increase in surfactants concentration. Furthermore, the percentage COD removal of dye is increased with the increase in surfactant concentration. The nature of solvent has minimal effect on percentage removal of dye. The ratio of solventto aqueous phase volume required for the removal of dye decreases with the increase in surfactant concentration. It is possible to back-extract dye into aqueous phase and recover solvent by using counterionic surfactants. The separation of aqueous phase from the aqueous-phase solvent dispersion is faster for amyl alcohol as compared to benzyl alcohol and methyl benzoate. A theoretical model based on ion-exchange reaction between surfactants and dye is used to analyze the experimental data. PMID- 15116853 TI - Landfarm performance under arid conditions. 1. Conceptual framework. AB - The primary disposal method for oily sludge in the Kingdom of Saudi Arabia, which is a major oil-exporting country in the world, is landfarming. It is an attractive method of oily sludge disposal in hot arid climatic conditions. Although landfarming technology was introduced to Saudi Arabia in 1982, no scientific studies have been conducted within the Kingdom to support this decision. The results presented in this paper are based on a comprehensive field experiment conducted under Saudi Arabian environmental conditions. Details of experimental setup and conceptual framework of degradation process based on field observations are presented in this paper. The paper also addresses kinetics of oily sludge degradation in landfarm cells under natural and enhanced conditions in the presence of water, nutrients, and tilling. The 12-month field study showed that weathering (evaporation) and not biodegradation is the overall dominant degradation mechanism occurring in landfarms in the study area. The results of this study showed that up to 76% of the oil and grease (O&G) in the sludge has been lost from soil as a result of weathering. However, the results of this study also indicated the primary mechanism for the loss of C17 and C18 alkanes as compared to branched alkanes was due to biodegradation. PMID- 15116852 TI - Enhanced accumulation of phosphate by Lolium multiflorum cultivars grown in phosphate-enriched medium. AB - Agricultural and animal husbandry practices combined with soil composition have caused phosphate overloading of farmlands in different parts of the U.S. and Europe. Movement of soluble phosphates (Pi) from phosphorus enriched soils results in degradation of natural aquatic systems, triggering serious environmental problems. Remediation of such sites using plants that tolerate and accumulate high concentrations of Pi in their aerial parts may be an attractive remediation technology. In the present study, Pi transport and accumulation potential of Marshall and Gulf ryegrass (Lolium multiflorum cultivars) were determined using a solution culture of seedlings. Ryegrass seedlings accumulated phosphorus (P) in excess of 2% of dry weight in their aerial parts when supplied with 5 g/L KH2PO4 in medium. Phosphorus accumulation was positively correlated with the concentration of phosphate (0-5 g/L KH2PO4) in medium. Plants grew well on medium containing 5 g/L KH2PO4, but concentrations above 5 g/L caused symptoms of toxicity. Scanning electron microscopy and energy-dispersive X-ray spectroscopy confirmed high P accumulation in different cell types of grass roots and shoots. Phosphate starvation and replenishment experiments point to the unique ability of these grasses to concentrate phosphate in the above-ground parts. It is hypothesized that the unique ability of these ryegrass cultivars may be due to the presence of efficient phosphate transport and sequestration mechanisms. PMID- 15116854 TI - Landfarm performance under arid conditions. 2. Evaluation of parameters. AB - Various parameters that influence the degradation processes in landfarming operations include moisture content, microbial density and composition, nutrients, and tilling. In this paper, a detailed evaluation of each parameter is presented based on field experiments. This paper also addresses kinetic of weathering, volatilization, and biodegradation mechanisms under natural attenuation and enhanced conditions including aeration in the soil pores by tilling; combined effects of tilling and nutrients in the soil; tilling and addition of waterto maintain a minimum moisture level; and combination of tilling, water, and nutrients. The effect of double loading under enhanced condition was also studied in this paper. The study shows that the tilling is very effective to to enhance the loss of oil and grease (O&G) fractions under arid conditions. However hydrocarbon loss in the absence of fertilizer and water was mainly due to weathering (volatilization). In the absence water and nutrients, the microbial counts were found to be low. The addition of water and fertilizer combined with the tilling helped in the significant reduction of O&G through both volatilization and biodegradation mechanisms with early reduction of n-alkanes through a biodegradation process followed by weathering. However, in order to determine the contribution of each of these two processes to the whole degradation, further work is required. The high loading rate resulted in retaining moisture content in the soil, and it delayed weathering and biodegradation. The high loading rate caused bacterial counts to increase, as it provided them with a plentiful source of food and water; however, it did not stimulate the biodegradation process for almost 6 months after the highest rate of sludge application to the soil. PMID- 15116855 TI - Hydrate composite particles for ocean carbon sequestration: field verification. AB - This paper reports on the formation and dissolution of CO2/seawater/CO2 hydrate composite particles produced during field experiments in Monterey Bay, CA using a CO2 injector system previously developed in the laboratory. The injector consisted of a coflow reactor wherein water was introduced as a jet into liquid CO2, causing vigorous mixing of the two immiscible fluids to promote the formation of CO2 hydrate that is stable at ambient pressures and temperatures typical of ocean depths greater than approximately 500 m. Using flow rate ratios of water and CO2 of 1:1 and 5:1, particulate composites of CO2 hydrate/liquid CO2/seawater phases were produced in seawater at depths between 1100 and 1300 m. The resultant composite particles were tracked by a remotely operated vehicle system as they freely traveled in an imaging box that had no bottom or top walls. Results from the field experiments were consistent with laboratory experiments, which were conducted in a 70 L high-pressure vessel to simulate the conditions in the ocean at intermediate depths. The particle velocity and volume histories were monitored and used to calculate the conversion of CO2 into hydrate and its subsequent dissolution rate after release into the ocean. The dissolution rate of the composite particles was found to be higher than that reported for pure CO2 droplets. However, when the rate was corrected to correspond to pure CO2, the difference was very small. Results indicate that a higher conversion of liquid CO2 to CO2 hydrate is needed to form negatively buoyant particles in seawater when compared to freshwater, due primarily to the increased density of the liquid phase but also due to processes involving brine rejection during hydrate formation. PMID- 15116856 TI - Characteristics of trapping copper ions with scrolled ferritin reactor in the flowing seawater. AB - Native liver ferritin of Dasyatis akajei (DALF), apoDALF, and reconstituted DALF were employed to construct a ferritin reactor, respectively. An apparatus consisting of a mixer, a ferritin reactor, and a magnetic stirrer was constructed to study capacity and feasibility of trapping Cu2+ in the flowing seawater. The experimental results showed that the numbers of trapping Cu2+ with DALF reactor were higher than these with the reactors of apoDALF and reconstituted DALF, respectively, giving the maximal numbers of 98 +/- 5 Cu2+ per molecular DALF in 120 h. We found that the iron layer with a high ratio of phosphate to ion on the surface of the ferritin core played an important role in increasing numbers of trapping Cu2+. In addition, we found two positive relations of dependence of trapping Cu2+ numbers with the reactor on the incubation time and on the Cu2+ concentration in the flowing seawater. Another apparatus consisting of a buoyage, an isolation basket equipped with griddling, and a scrolled ferritin reactor was constructed to study the feasibility of trapping Cu2+ in the sea area. Moreover, the present studies indicated that this apparatus had been used to not only analyze and evaluate the concentration variety of various heavy metal ions such as Cu2+ and Pb2+ diluting by the seawater but also monitor the formation of pollution degree by various small organic molecules during the climax and the neap. PMID- 15116857 TI - Influence of ion accumulation on the emulsion stability and performance of semi synthetic metalworking fluids. AB - The metalworking industry is one of the largest in the United States. Although metalworking fluids (MWFs) are ubiquitous in manufacturing as coolants and lubricants, these emulsified fluids have a significant environmental impact over their life cycle. Accordingly, it has become necessary to better understand emulsion destabilization mechanisms that lead to MWF deterioration and disposal so that MWF formulations can be designed for increased longevity. This paper investigates the impact of pH and a wide range of hard water salts on MWF emulsion stability. While expected trends from the emulsion science literature are observed, it is shown that MWF destabilization can lead to an increase in the microbial load that the MWF can sustain while only slightly improving manufacturing performance as measured by the tapping torque test. Experimental observations also indicate that these trends are strongly correlated with increased emulsion particle size, regardless of whether increased particle size is achieved by aging, by reductions in pH, or by the addition of hard water salts. In MWF systems, these conditions typically result from the accumulation of divalent and trivalent cations over time due to hard water additions and exposure to metal workpieces and tools. While MWFs are formulated with EDTA to avoid emulsion destabilization due to cation accumulation, it is shown that EDTA can be ineffective or highly inefficient for this purpose due to direct interactions between EDTA and the MWF emulsifier system. Given the ineffectiveness of EDTA and commonly utilized MWF emulsifier systems to maintain stable emulsion size in the presence of high concentrations of hard water salts, a more effective and environmentally preferable technological change to the MWF formulation design is proposed and successfully demonstrated. PMID- 15116858 TI - Experimental study of the adsorption of an ionic liquid onto bacterial and mineral surfaces. AB - Ionic liquids are being developed as a replacement for volatile organic solvents in a range of industrial applications. These liquids have a vanishingly small vapor pressure, making them an attractive alternative to the volatile organic solvents. However, a thorough assessment of the environmental impact of the use of ionic liquids requires a more complete understanding of their fate and transport in environmental systems. Toward this end, we measured the adsorption of the ionic liquid 1-butyl, 3-methylimidazolium chloride (Bmim CI) onto a range of surfaces meant to represent those commonly found in the near-surface environment. We measured adsorption onto the Gram-positive soil bacterial species Bacillus subtilis, onto gibbsite, onto quartz, and onto Na-montmorillonite. We conducted experiments as a function of pH, solid:solute ratio, time, and ionic strength. The experimental results reveal that Bmim CI is unstable in water below pH 6 and above pH 10 and that it exhibits pH independent and ionic strength dependent adsorption onto Na-montmorillonite with 0.4, 0.8, 1.0, 1.2, and 2.0 g/L of clay. We observed no adsorption of the Bmim CI onto B. subtilis (3.95 or 7.91 g (dry weight) bacteria/L) at pH 5.5-8.5 or onto gibbsite (500 or 1285 g/L) or quartz (1000 and 2000 g/L) over the pH range 6-10. Calculated distribution coefficient (KD) values for Bmim CI onto the Na-montmorillonite change as a function of ionic strength; the 10(-4) M ionic strength KD value is 1735 +/- 269 L/Kg, and the 10(-1) M ionic strength KD is 1133 +/- 291 L/Kg. Our results suggest that the geologic retardation of this class of ionic liquid, if present as a dissolved contaminant in the subsurface, would be significant when a significant fraction of interlayer clays are present. However, adsorption onto other common geologic and biological surfaces is likely to be minimal, and the ionic liquids may travel unimpeded in groundwater systems in which these types of surfaces dominate. PMID- 15116859 TI - Comment on "History and environmental impact of mining activity in Celtic Aeduan territory recorded in a peat bog (Morvan, France)". PMID- 15116860 TI - [Acute coronary syndromes without ST-segment elevation. From randomized clinical trials, to consensus guidelines, to clinical practice in Italy: need to close the circle]. AB - Recent therapeutic advances in the treatment of acute ischemic heart disease have been proven by randomized clinical trials and approved by formal practice guidelines. This rigorous approach has led to a sizable reduction in mortality and morbidity across the spectrum of acute coronary syndromes (ACS). However, contemporary registries of non-ST-elevation ACS set up by the cardiological community in Italy, as well as in the rest of Europe and in America, have shown only limited compliance to the general indication of treating high-risk patients by an early invasive approach protected by the use of glycoprotein IIb/IIIa receptor blockers. This partial failure in the process of improving patient care may be attributed to several reasons, including the suspect that practice guidelines may be biased by conflict of interest, concern about the applicability of the results of clinical trials to the real world, unrealistic expectations about treatment effects and, finally, logistic and economic obstacles including the availability of cath-labs and the high cost of platelet receptor blockers. Although the practice guidelines may provide a cultural support for translating the results of clinical research into patient care, and national and local cardiological associations can help in increasing awareness of the real benefits of an early aggressive approach in high-risk patients, the health care managers should remove bureaucratic obstacles and reallocate resources from treatments of unproven benefit to those that have been clearly shown to reduce mortality and the risk of reinfarction in ACS patients. PMID- 15116861 TI - [The global cardiovascular risk chart]. AB - BACKGROUND: Risk charts, built through risk functions deriving from longitudinal studies, are used in order to identify individuals at high risk for cardiovascular disease. For this reason the function has been identified and the global cardiovascular risk chart of the CUORE Project has been prepared, using Italian data coming from different cohorts enrolled between the '80s and the '90s, whose risk factors had been collected by standardized procedures. METHODS: The following risk factors have been used: age (10-year period, 40-49, 50-59, 60 69 years), gender (men and women), systolic blood pressure (< or = 129, 130-149, 150-169, > or = 170 mmHg), serum cholesterol (< or = 173, 174-212, 213-251, 252 290, > or = 291 mg/dl), smoking habit (yes, no) and presence of diabetes (yes, no); the first coronary or cerebrovascular event in people aged 40-69 years with no other previous cardiovascular events was considered as endpoint; survival has been assessed up to December 1998. RESULTS: Out of 18,028 people aged 40-69 years with no previous cardiovascular events, 647 first major cardiovascular events have been identified and validated, 449 coronary and 198 cerebrovascular. Charts are divided according to men and women and to diabetics and non-diabetics respectively; the different colors represent the percent level of risk and go from light green (< 5% in 10 years for men, < 1% in 5 years for women), dark green (between 5 and 10% for men, between 1 and 3% for women), yellow (between 10 and 15% for men, between 3 and 5% for women), orange (between 15 and 20% for men, between 5 and 7% for women), red (between 20 and 30% for men, between 7 and 10% for women), violet (> 30% for men, > 10% for women). CONCLUSIONS: The risk charts were built with data collected in recent years on men and women with a median follow-up of 10 years for men and 5 years for women, considering the first major fatal or non-fatal cardiovascular event as endpoint. Such a tool is easy to be applied by general practitioners and cardiologists in order to achieve a fast and objective evaluation of global cardiovascular risk. PMID- 15116862 TI - [Cardiac event recorder yields more diagnoses than 24-hour Holter monitoring in patients with palpitations]. AB - BACKGROUND: Palpitations are a common symptom that sometimes results from a substantial cardiac arrhythmia. A 24-hour Holter monitoring is usually used, but the yield of this instrument is low in patients whose symptoms occur infrequently. The aim of this study was to compare the diagnostic yield and the cost-effectiveness of transtelephonic event recorder (TER) with those of Holter monitoring in patients with intermittent palpitations. METHODS: Three hundred and ten patients with intermittent palpitations were allocated to the study and randomly assigned to receive a TER or 24-hour Holter monitoring. TER was given to patients until recording was obtained while symptoms occurred or was used at most for 7 days. At enrollment, a basal trace was recorded. Patients with palpitations recorded the one lead ECG trace and sent it by phone (fixed or mobile) to the telemedicine call center where a trained nurse compared the trace with the basal one and checked the patient's symptoms. The cardiologist reported "on-line" all the traces sent in the presence of an arrhythmic event and "stored and forwarded" all the other traces. Standard methods were used for Holter recording and reading. RESULTS: Patients with palpitations during the examination were 119 (76.8%) in the group of TER and 74 (47.8%) in the Holter group (p < 0.000) with an efficacy increase of 29% for TER. In symptomatic patients there were no differences between the two groups about the presence or absence of arrhythmias checked in the ECG traces; the time necessary to make a presence/absence diagnosis of arrhythmias was 2.97 +/- 2.74 days with the event recorder. The total cost of 155 tests made with Holter was altogether 9605.35 Euro (costs per test 61.97 Euro), while the one of TER was 6019.2 Euro (cost par test 38.83 Euro). The cost-effectiveness analysis was 129.80 Euro for Holter and 50.57 Euro for TER, with a saving of 79.23 Euro for every diagnosis made. CONCLUSIONS: TER allows to detect intermittent palpitations in real time; it is more useful and effective than Holter; moreover this effectiveness was also confirmed by the cost analysis in which TER resulted less expensive. PMID- 15116863 TI - [Telecardiology: perspectives and limitations]. PMID- 15116864 TI - [Concomitant treatment of atrial fibrillation in open-heart surgery patients: late rhythm and functional results]. AB - BACKGROUND: In recent years left atrial approaches have become popular because of technical simplicity, reproducibility and minor surgical trauma compared to the standard maze procedure. We analysed mid-term results of intraoperative left radiofrequency ablation of atrial fibrillation. METHODS: From February 1998 to August 2002, 206 patients (mean age 59.1 +/- 10.2 years) scheduled for open-heart surgery underwent combined treatment of atrial fibrillation at our Institution. In 163 patients (79.1%) a standard left epicardial approach was performed while 43 patients (20.9%) needed a totally endocardial approach. RESULTS: All patients but 3 underwent mitral valve surgery (104 repair and 99 replacement). Hospital mortality was 1.9%. Actuarial freedom from atrial fibrillation was 78% at 2 years and 74% at 4 years. At 4 years survival was 94% and freedom from neurological events was 98%. All patients with stable sinus rhythm 3 months after surgery recovered biatrial contractility. CONCLUSIONS: Left radiofrequency ablation of atrial fibrillation is effective in restoring sinus rhythm and normal atrial function. Combined atrial fibrillation treatment should be considered in all patients undergoing open-heart surgery. PMID- 15116865 TI - [A new approach to the closure of patent ductus arteriosus in premature infants]. AB - Minimally invasive surgical techniques have been employed successfully for a wide spectrum of cardiothoracic procedures. These approaches were proven to be as safe and effective as traditional surgical techniques. Unfortunately, it remains a lack of adequate instrumentations and anatomic features, particularly in the pediatric pool of patients. We have employed a new surgical approach for closing a patent ductus arteriosus in low weight newborn (< 2.5 kg), performing a superior extrapleuric approach. This report demonstrates that this technique is safe and reproducible, and is our opinion that it may substitute the medical management of patent ductus arteriosus. PMID- 15116866 TI - [Risk stratification after acute myocardial infarction: limitations of dobutamine stress echocardiography in females]. AB - BACKGROUND: Dobutamine stress echocardiography (DSE) is an imaging test widely used for risk stratification of patients after acute myocardial infarction. METHODS: We evaluated the sensitivity of DSE with respect to the gender and the stenotic coronary artery in patients who survived a myocardial infarction and with angiographic evidence of single-vessel coronary artery disease. RESULTS: The sensitivity of DSE was generally low. In particular, it was significantly lower in the presence of stenosis of the left circumflex and right coronary arteries with respect to the left anterior descending coronary artery. In females it was lower, especially when the stenosis involved the right coronary and left circumflex arteries. CONCLUSIONS: Our data suggest that in female gender the use of other imaging tests and particularly of coronarography should be strongly recommended for risk stratification after acute myocardial infarction. PMID- 15116867 TI - [New ways to become embroiled in a medico-legal suit. Encounters of the third type between cardiologists and lawyers in the era of guidelines and of molecular biology]. PMID- 15116868 TI - [Atrial septal defect in a child with Ito syndrome]. AB - Hypomelanosis of Ito is a rare neurocutaneous disorder described for the first time in 1952. It is characterized by depigmented skin areas often associated with ocular, musculoskeletal and neurological abnormalities. We report the case of a newborn affected by hypomelanosis of Ito with an atrial septal defect. PMID- 15116869 TI - [Type A acute aortic dissection presenting as mental status confusion: is it an atypically really uncommon presentation?]. AB - The most common initial symptom of acute aortic dissection is chest or abdominal pain. Nevertheless, in a minority of cases, it may have an atypical presentation, making the diagnosis clinically challenging. This article reports on a case of acute type A aortic dissection presenting as mental status confusion. The diagnostic suspicion for a cerebrovascular accident may have catastrophic consequences in a clinical condition, which is associated with a high mortality during the first 48 hours after the onset of symptoms if untreated. The right diagnosis was made by combining a careful physical examination, echocardiography, and computed tomography. The available literature about neurologic manifestations, their pathophysiology and prevalence as the initial symptom of acute type A aortic dissection is reviewed. PMID- 15116870 TI - [The use of a "transgastric oblique" transesophageal echocardiographic view to visualize the right ventricular inflow-outflow tract. Clinical applications]. AB - This paper describes the transesophageal echocardiographic "oblique transgastric" view to assess the right ventricular inflow-outflow tract. With this approach it is possible to view the right atrium, tricuspid valve, right ventricular inflow and outflow tracts, and the main pulmonary artery. In 2 clinical cases we evaluated the clinical feasibility of this approach in the diagnostic phase; in 31 patients who underwent cardiac transplantation we examined the advantages of this noninvasive hemodynamic monitoring. An advantage of transgastric view compared with esophageal standard views at 30-60 degrees, is the feasibility to obtain the velocity-time integral of the outflow tract systolic flow to calculate cardiac output by the pulsed Doppler sample volume orientation, which in this view is parallel to pulmonary arterial flow; moreover, it is possible to evaluate pulmonary artery pressures from pulmonary and tricuspid regurgitation. PMID- 15116871 TI - Studies on the pretreatment of zeolite clinoptilolite in packed beds. AB - The effect of volumetric flow rate, ranging from 5 to 45 Bed Volumes per hour (BV h(-1)) and temperature, ranging from 25 to 59 degrees C, during pretreatment of clinoptilolite on its effective capacity has been investigated. Pretreatment tests have been performed in an upflow ion exchange bed. Increased temperatures were found to increase the effective capacity of clinoptilolite. Effective capacity was maximal at low volumetric flow rates, indicating an influence of contact time and complete saturation of the zeolite bed at flow rates lower than 10 BV h(-1). Furthermore, a comparison between upflow and downflow operation at the same operating conditions showed that better results are obtained in upflow conditions, probably due to the better wetting of the material and the absence of liquid maldistribution. PMID- 15116872 TI - Mercury (II) removal from water by coconut shell based activated carbon: batch and column studies. AB - This study was undertaken to investigate adsorption behavior of Hg (II) from aqueous systems on activated carbon in static and dynamic mode by varying initial Hg (II) concentration, adsorbent dose and pH. Langmuir and Freundlich adsorption isotherm were applied to model the adsorption data. Removal of mercury obeyed the Langmuir and Freundlich adsorption isotherm models. The extent of removal of Hg (II) was found to be dependent on sorbent dose, pH and initial Hg (II) concentration. Mercury uptake increased from 72 to 100 percent with increasing pH from 2 to 10. A set of desorption studies was also performed for the metal ions with the aim of investigating the mechanism involved. Moreover, the competing effects of various ions like Pb (II) and Cu (II) is also described. The column capacity for a column diameter of 20 mm, bed height of 0.4 m, hydraulic loading rate of 7.5 m3 h(-1) m(-2) and a feed concentration of 3 mg l(-1) were found to be 3.02 mg g(-1). Breakthrough curves were plotted for the adsorption of mercury on the adsorbent using continuous-flow column operation by varying different operating parameters like hydraulic loading rate (3-10.5 m3 h(-1) m(-2)), bed height (0.3-0.5 m), and feed concentrations (2-6 mg l(-1)). The aim was to assess the effect of bed height, hydraulic loading rate and initial feed concentration on breakthrough time and adsorption capacity, which helped in ascertaining the practical applicability of the adsorbent. At the end an attempt has been made to develop empirical relationship from the data generated from column studies for designing the adsorption column, based on the Bohart-Adams model. PMID- 15116873 TI - Degradation of polycyclic aromatic hydrocarbons in soil: the Fenton reagent versus ozonation. AB - The ozonation and the Fenton treatment of soil spiked with a mixture of eleven polycyclic aromatic hydrocarbons (PAH) were studied. The efficiency of the treatment was strongly dependent on the matrix of soil (sand or peat). PAH adsorbed on sand undergo degradation more easily and require less oxidants (ozone, hydrogen peroxide) than PAH adsorbed on peat. Soil ozonation and the Fenton treatment were effective not only for the removal of 3-ring PAH, but could effectively degrade also 4-, 5- and more ring PAH. PAH removal from soil with the Fenton treatment in slurry was found to be dependent on the ratio of H2O2/soil/Fe2+, the manner of addition of hydrogen peroxide, and the treatment time. Three-phase ozonation of PAH contaminated soil resulted in a lower PAH removal and required higher ozone doses than two-phase ozonation. An improvement of the biodegradability during the chemical oxidation favours the implementation of combined chemical treatment and biodegradation for remediation of PAH contaminated soil. PMID- 15116874 TI - Full scale comparison of heterotrophic and nitrifying RBC biofilms. AB - Heterotrophic and nitrifying biofilms, from the secondary and tertiary full-scale Rotating Biological Contactor (RBC) wastewater treatment plants (WWTP) were investigated. Physical properties and structural features, such as, water content, organic portion (VS/TS), density, thickness, porosity, and fractal characteristics, including boundary/surface roughness, and Sierpinski and internal pore boundary fractals were examined. The results indicated that the heterotrophic biofilms were thicker and had higher porosity, larger internal voids, and faster growth and sloughing rates than the nitrifying biofilms. The nitrifying biofilms had much higher density(TS) and VS/TS ratio than the heterotrophic biofilms. In contrast to earlier results from laboratory scale studies, full-scale biofilms had structures and properties (porosity, density, and fractal dimensions) of more irregular spatial distribution. PMID- 15116875 TI - Ion trap LC/MS characterisation of toxic polar organic pollutants in colour photographic wastewaters and monitoring of their chemical degradation. AB - Liquid chromatography/electrospray ionisation-ion trap mass spectrometry (LC/ESI ITMS) with positive mode of operation was successfully applied to the characterisation of aromatic amines and chelating agents in colour photographic wastewaters. In addition to residual ingredients, monomers and dimers of sulphonated aromatic amines were the main toxic polar organic pollutants found. Oxidation of wastewater components by the Fenton-like reagent (Fe3+ + H2O2) was investigated by continuously pumping a solution of hydrogen peroxide. Iron concentration, present in the wastewater as ferric carboxylate complexes, was typically above 1 g l(-1), and therefore addition of Fe3+ was not necessary for treatment. Operating variables like reagent feeding concentration and flowrate, temperature and pH were studied. The overall chemical oxygen demand (COD) removal reached 90% after 7.5 h of treatment when the dosage of hydrogen peroxide was 230 g per litre of effluent, the pH was about 4 and the temperature was 60 degrees C. The absence of toxics in the treated effluents was confirmed by the Photobacterium phosphoreum luminescence reduction test. Monitoring of the chemical degradation of aromatic amines and chelating agents by LC/ESI-ITMS proved that the Fenton's like reagent was effective in degrading them. Propylenediamine tetraacetic acid (PDTA) was found to be the more recalcitrant compound, however about 97% of degradation was achieved after 7.5 hours of treatment. PMID- 15116877 TI - Effects of toxicants on nitrifying biomass in flocs vs. carriers. AB - Effects of chemical toxicants on nitrifying bacteria in flocs and porous carriers were compared. Mean inhibitory concentrations (IC50s) of toxicants were invariably higher in carrier suspensions than floc suspensions. The greatest differences were seen with cyanide, nickel and copper, whereas small differences were observed for the organic toxicants Methomyl and thiourea. Decreasing the size of the carriers lessened their protective effect. The results suggest that carriers would be advantageous for nitrification systems subjected to short-term inputs of heavy metals or cyanide but not helpful for systems subjected to transient inputs of organic chemicals. PMID- 15116876 TI - Removal of phosphorus from water using lignocellulosic material modified with iron species from acid mine drainage. AB - Lignocellulosic biosorbents, including juniper fiber, show promise as sorbents for removing pollutants from wastewater run-off because of their low cost. In this study, juniper fiber modified with iron species from acid mine drainage (AMD) was tested for its capacity to remove phosphorus from water compared to unmodified juniper fiber. In batch tests, the maximum adsorbate loading (Qmax) of phosphorus onto the modified adsorbent was 1.83 mg g(-1) at pH 4.0, obtained by fitting the isotherm results to the Langmuir isotherm model. This value is similar to the sorption capacity of other conventional adsorbents such as goethite, which implies that the modified lignocellulosic material would be effective as a sorbent for removing phosphorus from water. In the kinetic test, the pseudo-second order kinetic model fitted well the sorption of phosphorus onto the modified filter medium, showing the kinetic constant (k) of 8.09 x 10(-2) g(mg min)(-1) at initial phosphorus concentration of 10 mg l(-1). PMID- 15116878 TI - Aerobic biotransformation of octylphenol polyethoxylate surfactant in soil microcosms. AB - The biotransformation of octylphenol polyethoxylate surfactant (Triton X-100) and bacterial communities in soil microcosms was investigated. The soil microcosms were designed to simulate real sites of bioremediation. The soil used in this study was contaminated with pesticides and alkylphenol polyethoxylate surfactants over a long period of time. The nitrogen source, (NH4)2SO4 and the mineral salt basal solution were added to the microcosm and the water content was adjusted to 50% with distilled water. The microcosms were aerated using an aeration system with an air flow rate of 0.3 l min(-1). The exogenous bacterial strain Pseudomonas sp. SH4 was added to the microcosms to increase the rate of biotransformation of the Triton X-100. The number of microorganisms and the theoretical extent of formation of carbon dioxide were estimated to evaluate the biotransformation of Triton X-100 in the microcosms. The analytical results revealed that the exogenous bacteria could increase the rate of transformation of Triton X-100 by approximately 50%. Aeration of the microcosm increased the biotransformation of Triton X-100 by 45%. Bacterial count of 6.8x10(10) MPN g(-1) of soil was achieved in the M5 microcosm. Analysis of the bacterial community by 16S rDNA sequences revealed that Pseudomonas sp. SH4 could dominate all the microcosms to which it was added as an exogenous bacteria. The quantity of the indigenous bacterial strains Stenotrophomonas maltophilia, Stenotrophomonas sp. and clone 4-70 were also enhanced in the microcosms by the aeration, and the addition of carbon and nitrogen source. PMID- 15116879 TI - Nutrient removal in a sequencing batch biofilm reactor (SBBR) using a vertically moving biofilm system. AB - In this paper, the performance of a sequencing batch biofilm reactor (SBBR) using an anoxic/anaerobic phase followed by an aerobic phase for nutrient removal from wastewater was investigated. In the laboratory SBBR unit, biofilm was grown on a plastic biofilm media module, which was vertically moved up into the air and down into the bulk fluid during the aerobic phase. The vertical movement of the biofilm module supplied oxygen to the microorganisms. The module was submerged in the wastewater during the anoxic/anaerobic phase. The percentage removals of total chemical oxygen demand (COD), total nitrogen (TN) and soluble orthophosphate (PO4-P) were 95.4%, 68.3% and 89.5% respectively at influent concentrations of COD 773 mg l(-1), TN 49.9 mg l(-1) and PO4-P 16.5 mg l(-1). The effluent COD was 35 mg l(-1), NH4-N 10.2 mg l(-1), NO3-N 5.5 mg l(-1), soluble PO4-P 1.7 mg l(-1) and suspended solids (SS) 19 mg l(-1). PMID- 15116880 TI - Competitive adsorption of metals and organics onto a low cost natural polysaccharide. AB - Metals removal onto a low-cost natural polysaccharide is performed. Firstly, some specific characteristics of adsorbent are determined. Potentiometric titrations of the sorbent have been realised. The values of the point of zero net proton charge (pznpc) and the cation exchange capacity (CEC) deduced from these experiments have given respectively pH=5 and 0.575 meq g(-1). Secondly, sorption of cations from single metallic solution, have been performed and the equilibrium fixation capacities are 0.37 mmol g(-1) for Pb2+, 0.28 mmol g(-1) for Cu2+ and 0.2 mmol g(-1) for Ni2+. In multi-metals solutions of equimolar concentration, Ni2+ ions present the greatest fixation decrease in the presence of the two other cations (-61%), Pb2+ and Cu2+ seem to compete similarly. In the presence of a constant organic load composed of either benzaldehyde, benzoic acid or phenol and expressed as 100 mg l(-1) total organic carbon (TOC), benzoic acid induces the largest reduction of the copper equilibrium fixation capacity (-30%). PMID- 15116881 TI - Hydrogen-based tubular catalytic membrane for removing nitrate from groundwater. AB - A porous tubular ceramic membrane coated with palladium-cupper (Pd-Cu) catalyst on its surface was prepared and evaluated for catalytic reduction of nitrate from groundwater. Nitrate reduction activity and selectivity with the catalytic membrane were compared with Pd-Cu/Al2O3 catalyst particles. The catalytic membrane reactor exhibited a better selectivity by enabling an effective control of hydrogen gas, thus minimizing ammonium production. No leaching of palladium and copper into aqueous phase was observed, thereby indicating a high chemical stability of the metallic ions on the carrier support. This was also evidenced by the X-ray photoelectron spectroscopy (XPS) profiles of fresh and used catalysts, which showed no significant difference in surface compositions. Due to its higher selectivity in nitrate reduction and better flexibility in terms of operating conditions, the tubular catalytic ceramic membrane could be useful in removing nitrate from groundwater. PMID- 15116882 TI - Removal of metals in leachate from sewage sludge using electrochemical technology. AB - Heavy metals in acidic leachates from sewage sludge are usually removed by chemical precipitation, which often requires high concentration of chemicals and induces high metallic sludge production. Electrochemical technique has been explored as an alternative method in a laboratory pilot scale reactor for heavy metals (Cu and Zn) removal from sludge leachate. Three electrolytic cell arrangements using different electrodes materials were tested: mild steel or aluminium bipolar electrode (EC cell), Graphite/stainless steel monopolar electrodes (ER cell) and iron-monopolar electrodes (EC-ER cell). Results showed that the best performances of metal removal were obtained with EC and EC-ER cells using mild steel electrodes operated respectively at current intensities of 0.8 and 2.0 A through 30 and 60 min of treatment. The yields of Cu and Zn removal from leachate varied respectively from 92.4 to 98.9% and from 69.8 to 76.6%. The amounts of 55 and 44 kg tds(-1) of metallic sludge were respectively produced using EC and EC-ER cells. EC and EC-ER systems involved respectively a total cost of 21.2 and 13.1 CAN dollars per ton of dry sludge treated including only energy consumption and metallic sludge disposal. The treatment using EC-ER system was found to be effective and more economical than the traditional metal precipitation using either Ca(OH)2 and/or NaOH. PMID- 15116883 TI - Quantifying uncertainties in the assessment of sediment quality: statistical criteria and guidelines for sediment quality assessments. AB - Current sediment quality guidelines generally adopt a tiered approach in order to assess sediment quality more cost-effectively. The uncertainties involved in the tiered approach of an integrative assessment, however have not been quantified resulting in a risk of committing type I error or type II error at the final confirmatory stage. This study develops statistical criteria and guidelines for the sediment chemistry component of an integrative assessment of sediment quality. At the tier 1 screening stage, historical data or an initial survey is required to determine the minimum sample numbers that will be required to be representative of the study site. Understanding the guiding principles which have underpinned the setting of sediment quality criteria for contaminant is an important factor in tier 1 evaluations. To reduce cost and uncertainty in data, sampling should include the least number of samples necessary to minimise uncertainties by estimating the probability distribution function (which represents the variability of the natural environment at the location of concern), with subsequent application of kriging and sequential simulation methods on data obtained. Implementation of statistical criteria and guidelines in sediment quality assessments can provide a foundation for a further quantitative cost/benefit analysis and decrease the risk of committing type I and type II statistical errors at later classification stages. PMID- 15116884 TI - What is your diagnosis? Spondylosis of the lumbosacral junction. PMID- 15116885 TI - Low intensity heart murmurs in boxer dogs: inter-observer variation and effects of stress testing. AB - Inter-observer variation in the detection and grading of low intensity heart murmurs in boxer dogs was investigated. Six veterinarians with different levels of experience examined 27 boxers by cardiac auscultation. The dogs were auscultated before and after exercise, and the results were compared with phonocardiographic and echocardiographic examinations performed at rest and during two different stress tests. A subvalvular aortic ridge was identified in six dogs on two-dimensional echocardiography. Using dogs with low intensity murmurs or dogs free of heart murmurs, inter-observer agreement was positively correlated to the level of experience at rest (weighted kappa [kappa] 0.14 to 0.75), while the agreement was poor after exercise (weighted kappa 0.01 to 0.36). The presence of a subvalvular aortic ridge was associated with higher aortic flow velocities (P<0.002) and higher auscultatory murmur grading (P<0.001). There was an increase in murmur duration during one kind of stress test (P<0.001) and in aortic flow velocity during the other (P=0.001). PMID- 15116886 TI - Examination of serum total IgG1 concentration in atopic and non-atopic dogs. AB - In this study, serum immunoglobulin G1 (IgG1) concentrations were examined in atopic and non-atopic dogs receiving different levels of parasite control. Significantly lower serum total IgG1 concentrations were found in non-atopic dogs receiving stringent parasite control than in atopic dogs or non-atopic dogs receiving less stringent parasite control. Examination of serum total IgG1 concentrations of atopic dogs after six months of allergen specific immunotherapy (ASIT) showed a significant increase in serum total IgG1 concentrations. It is proposed that serum total IgG1 concentrations are affected by parasitism, atopic dermatitis and ASIT. PMID- 15116887 TI - Percutaneous fine-needle biopsy of deep thoracic and abdominal masses in dogs and cats. AB - Percutaneous fine-needle biopsy was used to investigate thoracic and abdominal masses in the dog and cat. One hundred and thirty-two cases were included in the study; 20 cases were excluded from the comparative study due to poor cellularity or blood contamination (retrieval rate 86.8 per cent). One hundred samples (56 dogs and 44 cats) were classified by cytology as neoplastic. All the cytological diagnoses of neoplasia were confirmed by histological samples obtained either by non-surgical methods, at surgery or during postmortem examination. No false positive diagnoses of neoplasia were made. Thirty-two samples were cytologically classified as 'negative for neoplasia'. Subsequent histological examination revealed 18 true negative and 14 false negative results. The procedure had an overall 89.4 per cent (118 cases out of 132) agreement between the diagnosis of inflammatory disease versus neoplasia, with a sensitivity of 87.8 per cent, a specificity of 100 per cent, a predictive value of a positive test of 100 per cent and a predictive value of a negative test of 56.3 per cent. PMID- 15116889 TI - Unusual presentations of cowpox infection in cats. AB - Cowpox virus infections are reported typically to cause focal ulcerated, crusted skin lesions, sometimes with mild systemic illness and concurrent oral lesions. Severe systemic illness usually only occurs in young or immunosuppressed individuals. This report describes four cases of cowpox infection in cats which illustrate variations to the usual presentation of the virus. The poxvirus infections were confirmed histopathologically, serologically and by PCR analysis. PMID- 15116888 TI - Acute paraplegia associated with vasculitis in a dog with leishmaniasis. AB - A 14-month-old female crossbreed dog with leishmaniasis, receiving allopurinol, was presented with acute paraplegia. A diagnosis of renal failure with pelvic limb lower motor neuron signs was made and the dog was euthanased. Histopathological examination demonstrated leukocytoclastic vasculitis in multiple organs. Malacia and haemorrhage affecting the spinal cord was associated with multiple foci of vasculitis within the nervous tissue. Rupture and thrombosis of inflamed vessels caused haemorrhage in the spinal cord and subsequent paralysis. PMID- 15116890 TI - Reversible pulmonary hypertension presenting simultaneously with an atrial septal defect and angiostrongylosis in a dog. AB - A one-year-old female neutered beagle was presented with marked abdominal effusion. Echocardiography showed marked dilatation of the right cardiac chambers, an atrial septal defect and severe tricuspid insufficiency. Systolic pulmonary arterial pressure (sPAP), evaluated by continuous wave Doppler echocardiography, was very high (80 mmHg), with a right to left interatrial shunt. The radiographic images were compatible with widespread pneumonitis. Numerous larvae of Angiostrongylus vasorum were visible on direct faecal examination. The animal was given fenbendazole for 15 days, combined with diuretics, an antibiotic and a vasodilator. Two weeks later, the dog showed a marked improvement. The treatment, except the anthelmintic, was continued for seven weeks and then stopped. At that stage, Doppler echocardiography revealed that the sPAP had returned to normal (20 mmHg) and the interatrial shunt had reversed (left to right). Eighteen months later, clinical and Doppler echocardiographic examinations were normal. PMID- 15116891 TI - Bilateral mandibular dentigerous cysts in a dog. AB - Cysts in the dental cavities of dogs are rare. This report documents the case of a young boxer that was referred after having had treatment for bilateral oral swellings. Dentigerous cysts were tentatively diagnosed. Surgery was performed to aspirate the fluid and remove the first premolars, which were positioned in the cysts. Histopathological examination of tissue collected during the surgical procedure confirmed the diagnosis. PMID- 15116892 TI - Pilot study to investigate the feasibility of surveillance of small animals in the UK. AB - This report presents a summary of the pilot Petsavers Companion Animal Disease Surveillance study that was initiated by the BSAVA to investigate the feasibility of veterinarians in practice collecting surveillance data from animals presented for treatment or routine procedures. Fifteen practices collected data using a standard questionnaire on up to four days over a period from July 2000 to April 2001. A total of 2631 questionnaires relating to first consultations were completed. There were errors in filling in the questionnaires and errors in data entry at a level that is standard for this type of data collection procedure. Some questions had a 30 per cent non-response rate. Approximately 60 per cent of the questionnaires contained data on dogs and 33 per cent on cats; other species were covered in the remaining 7 per cent of questionnaires. No questionnaires were returned for fish. Further analysis of the questionnaires indicated that 75 per cent of the dogs presented were pedigree, with 132 breeds listed, and 25 per cent were crossbreeds. The commonest disease group presented was ear disease: 4 per cent of dogs had one of five ear conditions. The conclusion from this pilot study is that a more robust technique for collection and preparation of data, that is less time consuming and more accurate, is required. PMID- 15116893 TI - Primary secretory otitis media in Cavalier King Charles spaniels. PMID- 15116894 TI - Practice standards: one scheme for all. PMID- 15116895 TI - Cutting edge of reptilian medicine. PMID- 15116896 TI - Sensitivity of finite helical axis parameters to temporally varying realistic motion utilizing an idealized knee model. AB - Various uses of the screw or helical axis have previously been reported in the literature in an attempt to quantify the complex displacements and coupled rotations of in vivo human knee kinematics. Multiple methods have been used by previous authors to calculate the axis parameters, and it has been theorized that the mathematical stability and accuracy of the finite helical axis (FHA) is highly dependent on experimental variability and rotation increment spacing between axis calculations. Previous research has not addressed the sensitivity of the FHA for true in vivo data collection, as required for gait laboratory analysis. This research presents a controlled series of experiments simulating continuous data collection as utilized in gait analysis to investigate the sensitivity of the three-dimensional finite screw axis parameters of rotation, displacement, orientation and location with regard to time step increment spacing, utilizing two different methods for spatial location. Six-degree-of freedom motion parameters are measured for an idealized rigid body knee model that is constrained to a planar motion profile for the purposes of error analysis. The kinematic data are collected using a multicamera optoelectronic system combined with an error minimization algorithm known as the point cluster method. Rotation about the screw axis is seen to be repeatable, accurate and time step increment insensitive. Displacement along the axis is highly dependent on time step increment sizing, with smaller rotation angles between calculations producing more accuracy. Orientation of the axis in space is accurate with only a slight filtering effect noticed during motion reversal. Locating the screw axis by a projected point onto the screw axis from the mid-point of the finite displacement is found to be less sensitive to motion reversal than finding the intersection of the axis with a reference plane. A filtering effect of the spatial location parameters was noted for larger time step increments during periods of little or no rotation. PMID- 15116897 TI - Is the wear factor in total joint replacements dependent on the nominal contact stress in ultra-high molecular weight polyethylene contacts? AB - The exact dependence of wear factor on contact stress, load and apparent contact area is much disputed in the literature. This study attempts to solve this dispute. Pin-on-plate studies of ultra high molecular weight polyethylene against stainless steel were conducted under different combinations of load (33-250 N), nominal stress (0.56-12.73 MPa) and face diameter, as well as two tests where both stress and load were kept constant, while the diameter was changed. For these tests the centre of the pin face was bored out to create four different average pin diameters with similar face areas. Diameter and load were found to have no significant effect on the wear factor, while the wear factor decreased with increasing contact stress according to the relation K = 2 x 10(-6) sigma( 0.84). PMID- 15116898 TI - An investigation into the effects of the hierarchical structure of tendon fascicles on micromechanical properties. AB - During physiological loading, a tendon is subjected to tensile strains in the region of up to 6 per cent. These strains are reportedly transmitted to cells, potentially initiating specific mechanotransduction pathways. The present study examines the local strain fields within tendon fascicles subjected to tensile strain in order to determine the mechanisms responsible for fascicle extension. A hierarchical approach to the analysis was adopted, involving micro and macro examination. Micro examination was carried out using a custom-designed rig, to enable the analysis of local tissue strains in isolated fascicles, using the cell nuclei as strain markers. In macro examination, a video camera was used to record images of the fascicles during mechanical testing, highlighting the point of crimp straightening and macro failure. Results revealed that local tensile strains within a collagen fibre were consistently smaller than the applied strain and showed no further increase once fibres were aligned. By contrast, between group displacements, a measure of fibre sliding, continued to increase beyond crimp straightening, reaching a mean value of 3.9 per cent of the applied displacement at 8 per cent strain. Macro analysis displayed crimp straightening at a mean load of 1 N and sample failure occurred through the slow unravelling of the collagen fibres. Fibre sliding appears to provide the major mechanism enabling tendon fascicle extension within the rat-tail tendon. This process will necessarily affect local and cellular strains and consequently mechanotransduction pathways. PMID- 15116899 TI - The application of force-sensing resistor sensors for measuring forces developed by the human hand. AB - Most attempts to measure forces developed by the human hand have been implemented by placing force sensors on the object of interaction. Other researchers have placed sensors just on the subject's fingertips. In this paper, a system is described that measures forces over the entire hand using thin-film sensors and associated electronics. This system was developed by the authors and is able to obtain force readings from up to 60 thin-film sensors at rates of up to 400 samples/s per sensor. The sensors can be placed anywhere on the palm and/or fingers of the hand. The sensor readings, together with a video stream containing information about hand posture, are logged into a portable computer using a multiplexer, analogue-to-digital converter and software developed for the purpose. The system has been successfully used to measure forces involved in a range of everyday tasks such as driving a vehicle, lifting saucepans and hitting a golf ball. In the latter case, results are compared with those from an instrumented golf club. Future applications include the assessment of hand strength following disease, trauma or surgery, and to enable quantitative ergonomic investigations. PMID- 15116900 TI - Development and experimental validation of a three-dimensional finite element model of the human scapula. AB - A new modelling approach, using a combination of shell and solid elements, has been adopted to develop a realistic three-dimensional finite element (FE) model of the human scapula. Shell elements were used to represent a part of the compact bone layer (i.e. the outer cortical layer) and the very thin and rather flat part of the scapula--infraspinous fossa and supraspinous fossa respectively. Solid elements were used to model the remaining part of the compact bone and the trabecular bone. The FE model results in proper element shapes without distortion. The geometry, material properties and thickness were taken from quantitative computed tomography (CT) data. A thorough experimental set-up for strain gauge measurement on a fresh bone serves as a reference to assess the accuracy of FE predictions. A fresh cadaveric scapula with 18 strain gauges fixed at various locations and orientations was loaded in a mechanical testing machine and supported at three locations by linkage mechanisms interconnected by ball joints. This new experimental set-up was developed to impose bending and deflection of the scapula in all directions unambiguously, in response to applied loads at various locations. The measured strains (experimental) were compared to numerical (FE) strains, corresponding to several load cases, to validate the proposed FE modelling approach. Linear regression analysis was used to assess the accuracy of the results. The percentage error in the regression slope varies between 9 and 23 per cent. It appears, as a whole, that the two variables (measured and calculated strains) strongly depend on each other with a confidence level of more than 95 per cent. Considering the complicated testing procedure on a fresh sample of scapula, the high correlation coefficients (0.89-0.97), the low standard errors (29-105 micro epsilon) and percentage errors in the regression slope, as compared to other studies, strongly suggest that the strains calculated by the FE model can be used as a valid predictor of the actual measured strain. The model is therefore an alternative to a rigorous three-dimensional model based on solid elements only, which might often be too expensive in terms of computing time. PMID- 15116901 TI - A kinematic method to calculate the workspace of the trapeziometacarpal joint. AB - The specific aim of this study was to develop a quantitative method and a kinematic method to evaluate the maximal workspace of the trapeziometacarpal (TM) joint. Six fresh-frozen human cadaver hands were disarticulated 4 cm proximal to the wrist joint and used in this experiment. The three-dimensional motion data of the TM joint was collected by an electromagnetic tracking device at 30 Hz. The workspace was reconstructed according to a complete set of motion data included circumduction, flexion-extension and abduction-adduction. A spherical fitting technique was used to obtain a sphere encompassing all the motion trajectories and estimating the centre of the sphere. The surface area of the maximal TM workspace, located on the one part of the sphere surface, was calculated by surface integration. The interclass correlation coefficient values for the reliability estimation of the repeated measurements of the radius and surface area of all specimens were 0.91 and 0.98 respectively. The mean coefficients of variance of the measured radius and the surface area were 2.04 per cent and 3.65 per cent respectively. The results also showed that using a spherical model to calculate the maximal workspace as an index for assessing TM joint impairment is practical. PMID- 15116902 TI - The benefits of using low accelerating voltage to assess endodontic instruments by scanning electron microscopy. AB - Scanning electron microscopy is often used to evaluate surface contamination and machining defects in dental and other medical instruments. Knowledge of the operating conditions of the SEM, in particular the accelerating voltage, is essential to properly interpret images of such material. We demonstrate the importance of using low accelerating voltages to detect surface features including contamination on NiTi rotary and hand files, and conclude that even recent studies may have significantly underestimated the amount of non-metallic debris (from the manufacturing process or from biological contamination) present on the surface of such instruments. PMID- 15116903 TI - Microbial aetiology of endodontic treatment failure and pathogenic properties of selected species. PMID- 15116904 TI - Unusual maxillary lateral incisors: case reports. AB - Root canal morphology is often complex and the number of root canals may vary for any type of tooth. Abnormalities in the root canal morphology of maxillary lateral inciors are rare. Maxillary lateral incisors can have two root canals, even though the dental literature supports their 100% single-canal anatomy. It is vital to consider the possibility of extra root canal(s), even in teeth with a low frequency of abnormal root canal anatomy. This report presents two cases of maxillary lateral incisors with two root canals. PMID- 15116905 TI - Evaluation of the antimicrobial effect of Er:YAG laser irradiation versus 1% sodium hypochlorite irrigation for root canal disinfection. AB - This laboratory study evaluated Er:YAG laser antibacterial action in infected root canals. Forty-eight maxillary central incisors were used. After canal preparation, the teeth were autoclaved and divided into four groups: (1) non treated teeth (control group); (2) teeth treated with NaOCl; (3) teeth irradiated with Er:YAG laser (7 Hz, 100 mJ, 80 pulses/canal, 11 sec) to the working length; (4) teeth irradiated similarly to, but 3 mm short, of the apex. The root canals from Groups 2, 3 and 4 were inoculated with 4 bacteria: Bacillus subtillus, Enterococcus faecalis, Pseudomonas aeruginosa, and Staphylococcus aureus, together with Candida albicans, and maintained for 24 h at 37 degrees C. All suspensions were adjusted to tube 2 of the MacFarland scale. The intracanal material was then collected with sterile paper points, which were placed in the canals for 5 min and then immersed in 5 ml of BHI medium. This was then seeded onto agar and stained by Gram's method. The NaOCl solutions and the Er:YAG laser irradiation to working length were effective against all five micro-organisms; however, 70% of the specimens irradiated 3 mm short of the apex remained infected. PMID- 15116907 TI - Clinical application of computer-aided rapid prototyping for tooth transplantation. PMID- 15116906 TI - The influence of filling technique on depth of tubule penetration by root canal sealer: a study using light microscopy and digital image processing. AB - The purpose of this study was to compare the depth of sealer penetration into dentinal tubules by three root-filling techniques using light microscopy and digital image processing. Thirty-two maxillary central incisors were prepared. Two teeth were separated for the control group. The rest were divided into three equal groups and obturated as following--G1: lateral condensation; G2: warm vertical compaction of gutta-percha and G3: Thermafil system. Each sample was sectioned longitudinally and prepared for microscopic analysis. A sequence of photomicrographs with magnifications of X50, X200 and X500 were taken. Through digital image analysis and processing, measurements for each field were obtained. A non-parametric ANOVA Kruskal-Wallis analysis was used to determine whether there were significant differences among the groups. Significant differences between G2 and G1 (p = 0.034) and between G3 and G1 (p = 0.021) were identified. There were no significant differences between G2 and G3 (p > 0.05). The results of this research suggest that samples root-filled by thermoplasticised gutta percha techniques lead to deeper penetration of the root canal sealer into the dentinal tubules. PMID- 15116908 TI - A letter from America. PMID- 15116909 TI - Capillary microextraction on sol-gel dendrimer coatings. AB - Sol-gel capillary microextraction (CME) is a new direction in the solventless sample preparation for the preconcentration of trace analytes, and presents significant interest in environmental, pharmaceutical, petrochemical, biomedical, agricultural, food, flavor, and a host of other important areas. It utilizes advanced material properties of organic-inorganic hybrid sol-gel polymers to perform efficient extraction and preconcentration of target compounds from a wide variety of matrices. In the present work, a novel benzyl-terminated dendron-based sol-gel coating was developed for CME. A detailed investigation was conducted to evaluate the performance of the newly developed sol-gel dendrimer coatings to perform solventless extraction of a wide range of polar and nonpolar analytes. The characteristic branched architecture of dendrons makes them structurally superior extraction media compared with their traditional linear polymeric counterparts. Sol-gel chemistry was used to chemically immobilize dendritic macromolecules on fused silica capillary inner surface. Due to the strong chemical bonding with the capillary inner walls, sol-gel dendron coatings showed excellent thermal and solvent stability in capillary microextraction in hyphenation with chromatographic analysis. Efficient extraction of a wide range of analytes from their aqueous solutions was accomplished using sol-gel dendron coated fused silica capillaries. Low parts per trillion level detection limits were achieved in CME-GC for both polar and nonpolar analytes including polyaromatic hydrocarbons (PAHs), aldehydes, ketones, phenols, and alcohols. PMID- 15116910 TI - Sequential ethoxycarbonylation, methoximation and tert-butyldimethylsilylation for simultaneous determination of amino acids and carboxylic acids by dual-column gas chromatography. AB - Amino acids (AAs) in alkaline solution were first ethoxycarbonylated with subsequent methoximation of keto acids (KAs). After acidification and solid-phase extraction, tert-butyldimethylsilylation was performed for direct analysis by gas chromatography (GC) on dual-columns with different polarities, which provided simultaneous separation of multiple amino acids, carboxylic acids (CAs) and keto acids, facilitating accurate peak confirmation based on matching with retention index sets characteristic of each analyte. The present method was linear (r2 > or = 0.9955) with good precision (0.1-9.4%) and accuracy (-8.6 to 9.9%), allowing simultaneous screening for diagnostic amino acids along with carboxylic acids and keto acids in urine from a phenylketonuria patient. PMID- 15116911 TI - Multiresidue determination of fluoroquinolones in milk by column liquid chromatography with fluorescence and ultraviolet absorbance detection. AB - Column liquid chromatography with fluorescence (FLD) and UV-diode array detection (UV-DAD) was used for the simultaneous determination of ciprofloxacin (CIPRO), enrofloxacin (ENRO), marbofloxacin (MARBO), danofloxacin (DANO) and sarafloxacin (SARA) residues in milk, using norfloxacin (NOR) as internal standard. Two solid phase extraction (SPE) cartridges, were evaluated for sample clean-up and preconcentration, Strata X, based on a modified styrene-divinylbenzene polymer, and Strata Screen A, a mixed anion exchanger/C8 reversed-phase sorbent. The fluoroquinolones (FQs) were separated on a polar endcapped column (AQUA C18). The recoveries for raw milk spiked with the antibiotics at three concentrations close to the maximum residue limit (MRL), were 80-103% for ENRO, CIPRO and DANO, with relative standard deviations (R.S.D.) lower than 6.6%. SARA recoveries were 70% (R.S.D. = 7%) and values in the order of 95% (R.S.D. = 1.5%) were obtained for MARBO at the MRL level. The quantification limits ranged from 2.4 to l0 ng ml(-1) and are below the MRL established for these drugs by the European Union. The method was successfully applied to the analysis of ENRO and its metabolite CIPRO in an incurred milk sample. PMID- 15116912 TI - Rapid multiresidue extraction method of organochlorinated pesticides from fish feed. AB - A rapid multiresidue extraction method for organochlorinated pesticides from fish feed was developed, which is based on the extracted fat treatment by n-hexane, concentrated sulphuric acid and ENVI-carb, a graphitized non-porous carbon material. The final residue, obtained in about 50 min, was dissolved in isooctane and analysed by gas chromatography with an electron capture detector (GC/ECD). The presence of the extracted pesticides was confirmed by gas chromatography-mass spectrometry (GC/MS). Concentration of sulphuric acid and amount of ENVI-carb were optimized in order to improve analytes recovery, accuracy and detection limits. This simple and relatively fast method allowed a high recovery of the HCB, Lindane, HEPO, p,p'-DDE, p,p'-DDD, p,p'-DDT residues, with mean values in the range 68-124% at four fortification levels (12.5, 25.0, 50.0, 100.0 ng/g), and coefficients of variation between 1.9 and 20.2%. Detection limit were equal to 3.0 ng/g, related to fat, for all pesticides, and calibration curves were linear (r > 0.999) in the range of explored concentrations from the detection limit to 100 ng/g. For all pesticides a good repeatability was obtained (CV% values in the range 0.23-4.16%) when a sequence of six injections of the isooctane extraction solution was performed. The usefulness of the proposed method has been tested by the analysis of fish feed samples. PMID- 15116913 TI - New insights on the retention mechanism of non-polar solutes in reversed-phase liquid chromatographic columns. AB - A clarification of the retention mechanism of non-polar solutes in octadecyl reversed-phase chromatographic columns is attempted based on a systematic comparison of the retention in C18 and C2 columns under the assumption that the retention in C2 columns is due to adsorption. The comparison involves curve fitting procedures and tests based on the properties of special functions suggested in the present paper. For the application of this approach the retention behaviour of six non-polar solutes, benzene, toluene, ethylbenzene, propylbenzene, isopropylbenzene and tert-butylbenzene, is studied from aqueous mobile phases modified with methanol, isopropanol, acetonitrile and tetrahydrofuran using C18 and C2 reversed-phase columns. It was found that the retention mechanism in C18 columns is not the same in the four modifiers. In particular, our results show that the adsorption mechanism has a significant contribution in mobile phases modified by acetonitrile and tetrahydrofuran, the partition mechanism is likely to predominate in isopropanol-water mobile phases provided that the mole fraction of isopropanol is higher than 0.2, whereas the case of MeOH is rather obscure, since the various tests did not give a clear picture about the retention mechanism in methanol-water mobile phases. PMID- 15116914 TI - Charge regulation in protein ion-exchange chromatography: development and experimental evaluation of a theory based on hydrogen ion Donnan equilibrium. AB - An extension of the stoichiometric displacement (SD) model for the ion-exchange adsorption of dilute proteins is developed which accounts for the effects of hydrogen ion Donnan equilibrium on the protein charge. The ability of the new model to fit retention data when the fluid phase pH is near the protein pI and the effects of hydrogen ion Donnan equilibrium are important is examined using four different proteins and four different column packings. The results indicate that the model is able to fit retention data using values for the protein pI and the change in protein charge with pH at the pI, i.e., (dz/dpH)pI, that are significantly closer to the values of these parameters determined by isoelectric focusing and acid-base titrametry in free solution, respectively, as compared to the values obtained by determining the characteristic binding change as a function of pH using the traditional stoichiometric displacement model. This suggests that when the fluid phase pH is near the protein pI, charge regulation is an important cause of the discrepancy between the electrical charge of a protein in free solution and the characteristic binding charge from the stoichiometric displacement model. The results also indicate that for the case where the fluid phase pH is near the protein pI, the new model accounts for the effect of charge regulation during protein ion-exchange adsorption more accurately than previous models in the literature. PMID- 15116916 TI - Molar mass distribution of a commercial aliphatic hyperbranched polyester based on 2,2-bis(methylol)propionic acid. AB - The determination of absolute molar mass averages (MMA) and molar mass distribution (MMD) of the fourth generation hyperbranched polyester Boltorn H40 (Perstorp Specialty Chemicals AB), synthesized from 2,2-bis(methylol)propionic acid (bis-MPA) as the AB2 monomer and ethoxylated pentaerythritol as the B4 core molecule was studied in dependence on the type of solvent, preparation procedure and solution concentration. Due to a large number of polar hydroxyl groups, ester, and also some residual carboxyl groups, a very stable H-bond network is formed at room temperature, that can-not be completely disrupted by dissolving the sample in solvents such as tetrahydrofuran (THF), N,N-dimethylacetamide (DMAc), a mixture of THF and methanol (9:1, v/v), and a solution of 0.7% LiBr in DMAc. The H-bonds between the polar groups break down completely and the dissolution of Boltorn H40 on the molecular level is achieved only when the sample is thermally pretreated at a minimum 140 C for at least 20 min prior to dissolution in solvents THF/methanol or LiBr/DMAc. Thus, determined MMA and molar mass distribution (MMD) of Boltorn H40 are independent on the kind of the solvent and solution concentration. PMID- 15116915 TI - Measurement and simulation of tailing zones of a cationic dye in analytical-scale reversed phase chromatography. AB - A quantitative physical description of tailing is reported here for analytical scale reversed phase chromatography with Type B silica. Simulations of experimental chromatograms for a cationic dye, 1,1'-didodecyl-3,3,3'3' tetramethylindocarbocyanine perchlorate (DiI) were performed as a function of DiI concentration and flow rate, revealing nonlinear tailing due to a bi-Langmuir adsorption isotherm. The strong site comprises less than 0.1% of the residual silanols, the desorption rate constant of DiI from this type of site is (85 +/- 8 ms)(-1), and the free energy of the silanophilic interaction is 16 kJ/mol, indicating hydrogen bonding or another strong electrostatic interaction. PMID- 15116917 TI - Comparison of chromatographic ion-exchange resins. III. Strong cation-exchange resins. AB - A comparative study was performed on strong cation-exchangers to investigate the pH dependence, efficiency, binding strength, particle size distribution, static and dynamic capacity, and SEM pictures of chromatographic resins. The resins tested included: SP Sepharose XL, Poros 50 HS, Toyopearl SP 550c, SP Sepharose BB, Source 30S, TSKGel SP-5PW-HR20, and Toyopearl SP 650c. Testing was performed with four different proteins: anti-FVII Mab (IgG), aprotinin, lysozyme, and myoglobin. Dependence of pH on retention was generally very low for proteins with high pI. An unexpected binding at pH 7.5 of anti-FVII Mab with pI < 7.5 was observed on several resins. Efficiency results show the expected trend of higher dependence of the plate height with increasing flow rate of soft resins compared to resins for medium and high-pressure operation. Determination of particle size distribution by two independent methods, Coulter counting and SEM, was in very good agreement. The mono-dispersed nature of Source 30S was confirmed. Binding to cation-exchange resins as a function of ionic strength varies depending on the specific protein. Generally, binding and elution at high salt concentration may be performed with Toyopearl SP 550c and Poros 50 HS, while binding and elution at low salt concentration may be performed with Toyopearl SP 650c. A very high binding capacity was obtained with SP Sepharose XL. Comparison of static capacity and dynamic capacity at 10% break-through shows in general approximately 50-80% utilisation of the total available capacity during chromatographic operation. A general good agreement was obtained between this study and data obtained by others. The results of this study may be used for selection of resins for testing in process development. The validity of experiments and results with model proteins were tested using human insulin precursor in pure state and in real feed stock on Toyopearl SP 550c, SP Sepharose BB, and Toyopearl SP 650c. Results showed good agreement with experiments with model proteins. PMID- 15116918 TI - Characterization of the selectivity of a phenytoin imprinted polymer. AB - The selectivity of analytical methods based on molecularly imprinted polymers (MIPs) is due to the preferential adsorption of the analyte(s) as compared to other substances (interferences). This paper shows the theoretical and practical difficulties, which have to be considered and solved when real samples need to be analysed in a wide range of analyte and interferant concentrations. It is shown that the estimation of interference effects requires either many measurements or a realistic model of the adsorption equilibrium in mixed solutions of the analyte and the interferences. Examples are shown for positive (cooperative) interference effects, for better experimental design and interpretation of binary isotherm measurements and for establishing the chemical model of interference from selectivity measurements. The usual MIP model consisting of a cavity, which closely fits the shape of the template from all sides, appears unsuitable for this MIP, and it is replaced with a more realistic, more open model. The applicability of the results to using non-imprinted polymers as selective sorbents and to screening drug candidates is also shown. PMID- 15116919 TI - Synthesis of chiral stationary phases with radical polymerization reaction of cellulose phenylcarbamate derivatives and vinylized silica gel. AB - Cellulose phenylcarbamate derivatives having methacrylate groups were synthesized with regioselective and non-regioselective procedures. These derivatives were chemically immobilized onto a vinylized silica gel, respectively, via a radical co-polymerization reaction. The immobilization was efficiently attained using a small amount of AIBN. The chiral recognition abilities of the prepared chiral stationary phases (CSPs) were evaluated by HPLC resolution of test enantiomers. It was observed that most of the enantiomers were completely resolved with markedly high column efficiency of 30,000-40,000 plates per metre for the eluted peaks. The effect of the amount of methacrylolyl chloride used for preparation on resolution was investigated. A direct comparison of the chiral recognition ability was made on the regioselectively and non-regioselectively prepared CSPs. In addition, the chemically bonded-type of CSPs were found to be relatively stable with addition of solvents such as tetrahydrofuran (THF) and chloroform into the mobile phase, which can lead to the dissolution of cellulose derivatives on the coated CSPs. Thus the choice of solvents used as the mobile phase is greatly extended and better resolution of several test enantiomers was observed on the prepared CSPs with THF and chloroform as a composition in the mobile phase. The batch-to-batch and run-to-run reproducibility was also discussed on the newly prepared CSPs. PMID- 15116920 TI - Comparison study of Chiralpak AD-H with AD columns in chromatographic enantioseparation of dihydropyrimidinone acid and its methyl ester. AB - This paper reports a comparison study of the difference between Chiralpak AD-H and AD columns in enantioseparation of dihydropyrimidinone (DHP) acid and its methyl ester under normal phase LC conditions. Unlike those of the AD phase, the van't Hoff plots of retention factors for DHP acid on the AD-H phase were linear. The cyclic van't Hoff plots of selectivity factors for DHP acid on the AD-H phase were non-linear and slightly non-superimposable. No conformational transition was observed on the AD-H phase in the whole temperature range. A single-step temperature program on the AD-H phase showed that the selectivity factors of DHP acid only increased approximately 1.7% in 24 h (versus approximately 50% on the AD phase). For DHP ester, the single-step temperature program showed that the selectivity factors on the AD-H phase remained the same in 24 h while those on the AD phase increased around 3.1%. The enantioselectivity of DHP acid on the AD H phase was lower than that on the AD phase while the enantioselectivity of DHP ester on the AD-H phase was higher than that on the AD phase. The resolution of DHP acid on the AD-H phase was about the same as that on the AD phase while the resolution of DHP ester on the AD-H phase was much higher than that on the AD phase. The results of DHP acid are opposite of what the vendor suggested while the results of DHP ester are the same as the vendor's application notes. This indicates that the differences between Chiralpak AD-H and AD columns are not only in their particle size, but also in the solvated conformations. PMID- 15116921 TI - Characterization of implant device materials using size-exclusion chromatography with mass spectrometry and with triple detection. AB - A more complete understanding of the raw materials used for making implant device materials becomes increasingly important in the medical device industry. Often such detailed information requires utilization of a combination of analytical techniques. In this work, we characterize a poly(dimethyl siloxane) (PDMS) material using on-line size exclusion chromatography (SEC) with electrospray ionization (ESI) and matrix assisted laser desorption ionization (MALDI) mass spectrometry (MS) techniques. Here, we obtain detailed molecular compositional information such as repeat units, end group chemistry, and identification of impurities in both the high and low mass range. SEC with light scattering, viscosity, and refractive index detection (triple detection) is used to obtain information on a small quantity of high mass impurity that was undetected by both SEC-ESI and MALDI MS techniques. SEC with triple detection measures absolute molecular weights and molecular weight distributions. We compare average molecular weight values of the implantable device polymer obtained by SEC with triple detection, SEC-ESI, and SEC-MALDI MS techniques. PMID- 15116923 TI - Rapid analysis of nucleotide-activated sugars by high-performance liquid chromatography coupled with diode-array detection, electrospray ionization mass spectrometry and nuclear magnetic resonance. AB - A generally applicable method for HPLC analysis of sugar nucleotides was established. Separation was achieved using ion-pair chromatography on a reversed phase column. Ion-pair reagents were selected and various parameters optimized with respect to separation of 11 of the most important sugar nucleotides and compatibility with on-line detection by electrospray ionization MS and NMR. The method was applied to the on-line analysis of the GDP-D-mannose-4,6-dehydratase (Gmd) and GDP-4-keto-6-deoxy-D-mannose reductase (Rmd) catalyzed conversion of GDP-D-mannose to GDP-D-rhamnose. By LC-NMR, the intermediate product of the reaction was shown to be a mixture of GDP-4-keto-6-deoxy-D-mannose and GDP-3-keto 6-deoxy-D-mannose. Nucleotide co-factors of enzymatic reactions such as ATP and NADH did not interfere with the analysis of nucleotide-activated sugars. PMID- 15116922 TI - Separation and detection of oxidation products of fluorodeoxyglucose and glucose by high-performance liquid chromatography-electrospray ionisation mass spectrometry. AB - 2-Deoxy-2-[18F]fluoro-D-glucose ([18F] FDG), the most popular positron emitting radiopharmaceutical, may oxidise by autoradiolysis in aqueous solution. The aim of this work was to use LC-MS for determination of the oxidation products of fluorodeoxyglucose and glucose (Glc) obtained by oxidation with Fenton's reagent. Asahipak NH2P-50 polyamide silica column and acetonitrile-0.025% aqueous ammonium formate (80:20 (v/v)) eluent were utilised with an Agilent 1100 HPLC-MS instrument. Ten major oxidation products of FDG and Glc were separated and identified by mass spectrometry: 2-fluorogluconic acid, 2-fluoroglucuronic acid, 2-oxoerythronic acid, arabinose, arabonic acid, araburonic acid, erythrose, erythrulose, gluconic acid, and glucuronic acid. The most intensive electrospray ionisation signals were found in the negative ion spectra and were due to HCOO- adducts, the other acids being in their lactone forms. PMID- 15116924 TI - Purification of the precursor for the automated radiosynthesis of [18F]FCWAY by counter-current chromatography. AB - Radiolabeled FCWAY (N-(2-[4-(2-methoxyphenyl)piperazino])-N-(2-pyridinyl) trans-4 fluorocyclohexanecarboxamide) was prepared for human positron emission tomography (PET) studies by a simple one-step radiosynthesis. The LC-MS analysis of the products indicated that it contained impurities which may interfere with FCWAY uptake of 5-HT1A receptors and that these impurities were derived from an impurity originally present in the precursor preparation. Since preparative HPLC failed to resolve one of the impurities from the precursor, preparative-scale high-speed counter-current chromatography (HSCCC) was used for purification of this FCWAY precursor. A suitable two-phase solvent system composed of cyclohexane ethyl acetate-methanol-water at a volume ratio of 1:1:1:1 or 4:5:4:5 was selected based on the partition coefficients of the precursor and impurity as determined by a LC-MS method. Using the second solvent ratio of 4:5:4:5 with the organic phase as a mobile phase, a 2.57 g amount of precursor preparation was successfully purified yielding 2.2 g of the pure precursor by a single run. PMID- 15116925 TI - Ion exchange using poorly activated supports, an easy way for purification of large proteins. AB - Ion-exchange chromatography using commercial ionic supports is a commonly used technique for protein purification. However, selective adsorption of a target protein from a given extract onto commercial ion exchangers seems to be quite complex since they are designed to adsorb the maximum percentage of proteins with the opposite charge. In this paper, ion-exchanger supports with different activation degrees (from 1 to 40 micromol of amino groups per g of agarose) have been prepared and used for the purification of large proteins. These kinds of proteins have large surfaces to interact by many points with the support. Therefore, it was possible to purify large proteins as beta-galactosidase from Thermus sp. strain T2 from a crude extract from Escherichia coli or bovine liver catalase from a commercial preparation, with tailor-made ion-exchanger supports. A simple step of adsorption/desorption on lowly activated supports rendered both enzymes rather pure as confirmed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Moreover, this strategy makes also easy the desorption step that requires rather low NaCl concentrations, which may become a serious problem for desorption of large proteins when using conventional supports, due to their ability of generating a very strong adsorption. PMID- 15116926 TI - Enantioseparation of four cis and trans diastereomers of 2',3'-didehydro-2',3' dideoxythymidine analogs, by high-performance liquid chromatography and capillary electrophoresis. AB - Compounds 1-4 are the four stereoisomers of a synthetic new potential antiviral agent (d4T analog) containing two chiral centers and a base (uracil). Both high performance liquid chromatography (HPLC) and capillary electrophoresis (CE) techniques were used to separate and quantify enantiomers with high resolution. The determination of enantiomeric purity of the compounds was developed using both amylose chiral stationary phase by HPLC and anionic cyclodextrins (highly S CD) as chiral selectors in CE. The HPLC method was found to be superior in sensitivity to the CE method. PMID- 15116927 TI - Determination of the adsorption model of alkenes and alcohols on sulfonic copolymer by inverse gas chromatography. AB - The determination of a number of adsorption sites on sulfonated styrene divinylbenzene copolymer for alkenes (propene, 1-butene, 1-pentene, 1-hexene, 1 heptene, isobutene, 2-methyl-1-butene, 2-methyl-2-butene, 2-methyl-1-pentene, 2 methyl-2-pentene and 2-methyl-2-hexene) and alcohols (methanol, ethanol and n propanol, n-butanol, 2-butanol and tert-butanol) was performed by the saturation copolymer with vapors of adsorbate, by removing the excess of adsorbate from copolymer by blowing the inert gas through copolymer bed and by the desorption of adsorbed alcohol in the programmed increase of temperature. The adsorption measurements were performed on sulfonated ion-exchange resin (Amberlyst 15) with different concentrations of the acid group, which means with a varying number of adsorption sites. The following adsorption models for alkenes were suggested: the first in which one molecule of alkene is adsorbed by two sulfonic groups, for linear alcohols, the second in which one sulfonic group can adsorb one molecule of alcohol and for non-linear alcohols the third where one molecule of alcohol is adsorbed by two or more sulfonic groups. PMID- 15116928 TI - Identification of 2,3-butanedione (diacetyl) as the compound causing odor events at trace levels in the Llobregat River and Barcelona's treated water (Spain). AB - A study of organic compounds imparting sweet and buttery odor problems in the Llobregat River (northeast Spain) and in treated water was conducted. Solid-phase microextraction (SPME), gas chromatography-olfactometry, and flavor profile analysis (FPA) were used as analytical methodologies to identify the compound responsible for odor incidents. 2,3-Butanedione (diacetyl) with a concentration range of 0.90-26 microg/l in river water samples entering the water treatment plant was identified as the compound causing the odor events. Flavor profile analysis establishes 0.05 microg/l as its odor threshold concentration (OTC) in water, with an odor recognition concentration of 0.20 microg/l. The analyses were carried out with SPME-GC-MS and parameters affecting SPME extraction such as selection of the fiber (carboxen-polydimethylsiloxane), extraction time (30 min), temperature (60 degrees C), and ionic strength were evaluated. Quality parameters of the optimized method gives good linearity (r2 > 0.999), a limit of detection (0.08 microg/l) similar to the OTC of the compound, and good reproducibility (R.S.D. < 20%). The SPME method was applied to identify the compound causing the odor. PMID- 15116929 TI - Characterisation of different clones of Picea abies (L.) Karst using head-space sampling of cortical tissues combined with enantioselective capillary gas chromatography for the separation of chiral and non-chiral monoterpenes. AB - Head-space sampling (HS) has been combined with enantioselective gas chromatography (GC) for the analysis of chiral and non-chiral monoterpenes present in the cortical tissues of five different Norway spruce clones. (1S)-(-) alpha-Pinene, (1S,5S)-(-)sabinene, (1S)-(-)-beta-pinene, and (4S)-(-)limonene dominated over (1R)-(+)-alpha-pinene, (1R,5R)-(+)-sabinene, (1R)-(+)-beta-pinene, and (4R)-(+)-limonene. Results showed a large variation in the enantiomeric composition of cortical tissues between different clones. The development of HS GC greatly increased the speed of precise analyses of chiral monoterpenes in small samples and therefore offer excellent opportunities in studies on the ecophysiological and chemotaxomic roles of these chiral components. PMID- 15116930 TI - Multielemental speciation analysis of organometallic compounds of mercury, lead and tin in natural water samples by headspace-solid phase microextraction followed by gas chromatography-mass spectrometry. AB - The development of a simple and rapid multielemental speciation method is described with the ultimate goal to simultaneously determine various organometallic compounds of mercury, lead and tin (inorganic mercury, methylmercury, trimethyllead, triethyllead, monobutyl-, dibutyl- and tributyltin) in natural water samples. The analytical method consists on the ethylation with NaBEt4, simultaneous headspace-solid phase microextraction (HS-SPME) of the derivatives and final gas chromatographic-mass spectrometric (GC-MS) analysis. After optimization of important process parameters, like SPME fiber coating, extraction time and extraction temperature, the analytical characteristics were evaluated. Detection limits in the low ng l(-1) level, linearity over three orders of magnitude and repeatability in the range of 3-20% were achieved for all compounds under study. The accuracy of the method in terms of average percentage recovery of the compounds in spiked river water and seawater samples was better than 90%. Finally, application of the proposed method to real natural aqueous samples enabled the simultaneous determination of all the compounds under study in seawater samples obtained from the marina area of Gijon (Asturias, Spain). PMID- 15116931 TI - Application of comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry in the analysis of volatile oil of traditional Chinese medicines. AB - This paper reports comprehensive two-dimensional gas chromatography-time-of flight mass spectrometry (GC x GC-TOF MS) analysis of Pogostemon cablin Benth (Cablin Parchouli) volatile oil. The suitable column system and operation conditions were chosen on the basis of the properties of composition of the volatile oil. One-dimensional gas chromatography (ID-GC) and GC x GC, GC-MS and GC x GC-TOF MS were compared under appropriate conditions, and the enhanced sensitivity and superior resolution of GC x GC were demonstrated. 394 components were tentatively identified by GC x GC-TOF MS. PMID- 15116932 TI - Separation of derivatized alcohol ethoxylates and propoxylates by low temperature packed column supercritical fluid chromatography using ultraviolet absorbance detection. AB - Supercritical fluid chromatography (SFC) is capable of separating oligomers of alcohol ethoxylates (AEOs) and propoxylates (APOs) samples with pure carbon dioxide. The instrumental conditions, however, needed for separation necessitate both high temperature and high pressure. Derivatization of alcohol polyether samples with an UV absorbing agent has been achieved with a phenylated disilazane in hopes of employing a solvent-modified CO2 mobile phase in conjunction with both lower CO2 pressure and lower temperature for oligomer separation. A silylether containing a single phenyl group was formed via the derivatization of the hydroxyl termini of AEO and APO samples. The derivatized polyethers were detected at 215 nm with little or no interference from the mobile phase. Octadecylsilica (ODS) and a polar embedded alkyl bonded silica stationary phase were studied with the organic solvent-modified CO2 mobile phase. The combination of an ODS phase and the polar embedded phase, tandemly stacked, produced the best chromatographic separation of oligomeric species. Data from SFC-UV separations combined with peak assignments from SFC with electrospray ionization-mass spectrometric (ESI-MS) detection produced average molar oligomer values for each surfactant sample. PMID- 15116933 TI - Separation of quaternary ammonium diastereomeric oligomers by capillary electrophoresis. AB - The separation of novel diastereomeric trimers (3M) and pentamers (5M), derived from quaternary ammonium salts, was studied in conventional, uncoated and coated capillaries using capillary zone electrophoresis (CZE) with a variety of buffers and additives. Resolution of 5M diastereomers was best achieved using gamma cyclodextrin (gamma-CD) as a chiral selector, while no diastereomeric resolution was realized for the 3M material. PMID- 15116935 TI - Use of dynamically coated capillaries for the determination of heroin, basic impurities and adulterants with capillary electrophoresis. AB - Rapid, precise, accurate, and reproducible methodology using capillary electrophoresis (CE) with dynamically coated capillaries for the analysis of heroin and its basic impurities and adulterants is presented. Highly selective determination of the above solutes is obtained by analyzing the same sample preparation by two CE methods. For the determination of heroin, its basic impurities and basic adulterants, dynamic coating of the capillary surface is accomplished using a commercially available reagent kit with an added cyclodextrin ((CD) polycation coating followed by polyanion coating with dimethyl beta-cyclodextrin or hydroxypropyl-beta-cyclodextrin). The addition of a cyclodextrin to the run buffer significantly improves the separation of these solutes. Neutral, acidic, and weakly basic adulterants which migrate near or after t0 do not interfere with the more mobile basic solutes. The determination of neutral, acidic, and weakly basic adulterants in heroin is accomplished using a modification of the above commercially available reagent kit. After first coating with a polycation, a negative coating is obtained using a surfactant sodium dodecyl sulfate. Micellar electrokinetic chromatography (MEKC) with dynamically coated capillaries gives an excellent separation of the neutral, acidic, and weakly basic solutes, with considerably shorter run times compared to conventional MEKC. In addition for this system, most basic solutes in heroin have longer migration times than the uncharged and acidic compounds. PMID- 15116934 TI - Separation of ethoxylated bisphenol A dimethacrylates in dental composite after derivatisation to ionisable amines by capillary zone electrophoresis. AB - Bisphenol A ethoxylate dimethacrylates (Bis-EMA) are transformed into ionisable amines by derivatisation in order to make the analytes applicable to capillary electrophoresis. For this goal, piperidine was added onto the C=C double bond of the alpha,beta-unsaturated ester group forming a tertiary amine with pKa values between 9 and 10. Formation of the derivatives was confirmed by electrospray ionisation MS. Commercial Bis-EMA is a mixture of homologues with different number of ethoxy groups; it is characterised by the average number of the ethoxy groups in the chains. These homologues were resolved by capillary zone electrophoresis at pH 4. It is shown for the product with an average of four ethoxy groups per Bis-EMA molecule that about seven homologues can be baseline separated when differing by only one ethoxy group. For Bis-EMA with 30 ethoxy groups in average, about 23 homologues could be differentiated. The high resolution power of capillary zone electrophoresis enables characterisation of commercial dental composite material concerning the Bis-EMA constituents. PMID- 15116936 TI - Ultrasound-assisted Soxhlet extraction: an expeditive approach for solid sample treatment. Application to the extraction of total fat from oleaginous seeds. AB - Conventional Soxhlet extraction assisted in the cartridge by ultrasound has been developed and used to extract the total fat content from oleaginous seeds such as sunflower, rape and soybean seeds. The application of ultrasound to the sample cartridge enormously decreases the number of Soxhlet extraction cycles needed for quantitative extraction of the fat, thus reducing the extraction time at least to half the time needed by the conventional procedures. The results agree well with those obtained by conventional Soxhlet extraction and the ISO reference method, both in terms of efficiency and precision. The repeatability of the proposed approach, expressed as relative standard deviation, was 0.9%; the within laboratory reproducibility was 1.3%. Qualitative analysis of the extracted fat showed that the application of ultrasound does not change the composition of the oil. PMID- 15116937 TI - Determination of epichlorohydrin by sulfite derivatization and ion chromatography: characterization of the sulfite derivatives by ion chromatography mass spectrometry. AB - This work is an upgrade of a previously developed method (J. Chromatogr. A 884 (2000) 251] for epichlorohydrin determination by ion chromatography (IC) and conductivity detection. Here, an ion chromatography-mass spectrometry (IC-MS) coupling has been employed for the separation and the identification of products of epichlorohydrin when reacted with the nucleophilic agent SO3(2-). The high capacity column (IonPac AS11-HC) used for separation provided good resolution. This allowed evaluation of the IC behavior and mass spectrometric identification of epichlorohydrin sulfite derivatives. By using atmospheric pressure interfaces (ESI and APCI) the following species were tentatively identified: 2,3-dihydroxy-1 propanesulfonic, 2,3-epoxy-1-propanesulfonic,1,3-dihydroxy-2-propanesulfonic and 3-oxetanesulfonic acids and 2-hydroxy-1,3-propanedisulfonic acid (or its isomer 3 hydroxy-1,2-propanedisulfonic acid). The study showed that chlorine atoms are displaced from epichlorohydrin during the reaction, while mass spectrometry confirmed that none of the products formed contains chlorine atoms. PMID- 15116938 TI - Methods for the analysis of cannabinoids in biological materials: a review. AB - Various methods for the analysis of cannabinoids in biological materials, including plant and human body materials, are reviewed. Chromatographic methods, such as TLC, GC and HPLC, and non-chromatographic methods, mainly immunoassays, are discussed and compared. Chromatography is most commonly used in the analysis of plant material, with GC apparently offering the most advantages. Immunoassays, such as radioimmunoassay and fluorescence polarisation immunoassay, and enzyme immunoassay methods, such as enzyme multiplied immunoassay technique and enzyme linked immunosorbent assay, can be used for human body materials; however, GC-MS is still necessary for confirmation and accurate quantification. Preferred methods are suggested for various specific purposes. PMID- 15116939 TI - Qualitative analysis and HPLC isolation and identification of procyanidins from Vicia faba. AB - The soluble proanthocyanidins of the coloured seed coats of Vicia faba L. were isolated and separated by solvent partition. The chemical characteristics of the proanthocyanidins were elucidated by total oxidation and partial degradation in the presence of phloroglucinol followed by HPLC analysis. The native extract of proanthocyanidins contained (+)-gallocatechin, (-)-epigallocatechin, (+)-catechin and (-)-epicatechin units. Oligomeric procyanidins were purified by chromatography on Sephadex LH-20 and the accessible compounds were isolated by RP HPLC using a Licrospher Li 100 Column. The structures of the purified oligomeric procyanidins were elucidated using a procedure involving TLC, UV spectroscopy, ESI-MS and HPLC analysis of the products from the phloroglucinol reaction. The major condensed tannins of Vicia faba comprise six compounds identified as two A type procyanidin dimers, the procyanidin dimers B1, B2 and B3, and a procyanidin trimer. PMID- 15116940 TI - Methods of phytochemical standardisation of rhizoma Cimicifugae racemosae. AB - The methanolic extract of roots of Cimicifuga racemosa and its methanolysis products have been analysed by GC-MS. 2-Hexylcyclopropaneoctanoic acid (9,10 methylenehexadecanoic acid) was found to be the marker most specific for the identification for this herb. For the phytochemical standardisation of Rhizoma Cimicifugae racemosae, validated methods for the quantitative analysis of formononetin (by TLC-fluorometry), of isoferulic acid (by GC-MS of the methyl ester), and of total triterpene glycosides (transformed to coloured complexes and measured photometrically) in roots of C. racemosa have been developed. The contents of formononetin, isoferulic acid and total triterpene glycosides (measured as actein) in the herb ranged from 0.0031 to 0.0035, from 1.22 to 1.35 and from 20.09 to 22.06 mg/g dry weight, respectively. PMID- 15116941 TI - The determination of n-alkanes in the cuticular wax of leaves of Ludwigia adscendens L. AB - An n-hexane extract of fresh, mature leaves of Ludwigia adscendens, containing a thin layer of epicuticular waxes, has been analysed for the first time by TLC, IR and GC using standard hydrocarbons. The leaves contained 22 identified long chain (C15-C36) n-alkanes, accounting for 74.27% of the hydrocarbons present, and an unknown number of unidentified branched chain alkanes. The predominant n-alkane was C25 (11.02%), whilst C18 (7.62%), C20 (6.14%), C29 (5.36%) and C27 (5.29%) n alkanes were moderately abundant: the C35 homologue was present only in minor amounts (0.22%). PMID- 15116942 TI - Development of a one-step immunochromatographic strip test for the detection of sennosides A and B. AB - An immunochromatographic strip test was developed to detect sennoside A (1) and sennoside B (2) using anti-1 and anti-2 monoclonal antibodies. The qualitative assay was based on a competitive immunoassay in which the detector reagent consisted of colloidal gold particles coated with the respective sennoside antibodies. The capture reagents were 1- and 2-human serum albumin (HSA) conjugates immobilised on a nitrocellulose membrane on the test strip. The sample containing 1 and 2, together with detector reagent, passed over the zone where the capture reagents had been immobilised. The analytes in the sample competed for binding to the limited amount of antibodies in the detector reagent with the immobilised 1- and 2-HSA conjugates on the membrane and hence positive samples showed no colour in the capture spot zone. Detection limits for the strip test were 125 ng/mL for both sennosides. The assay system is useful as a rapid and simple screening method for the detection of 1 and 2 in plants, drugs and body fluids. PMID- 15116943 TI - Identification of the major vanilloid component in Capsicum extract by HPLC-EC and HPLC-MS. AB - A sensitive multi-channel HPLC-electrochemical (EC) method has been developed to determine the vanilloid content in the complex Capsicum annuum extract Capsibiol. Chromatographic separation was achieved within 10 min using a YMC Basic S5 column with a mobile phase containing chloroacetic acid, heptane sulphonic acid and acetonitrile. The multi-channel detector simultaneously applied four potentials between +500 and +800 mV (referenced to a silver/silver chloride electrode) to four glassy carbon working electrodes. The most abundant (0.94 mg/g) vanilloid analogue in the Capsibiol sample demonstrated an electrochemical reactivity and retention time similar to that of vanillic acid in HPLC-EC analysis. Its identity was confirmed by HPLC-MS using a Zorbax SB-CN column with a mobile phase containing formic acid and methanol. PMID- 15116946 TI - Current awareness in phytochemical analysis. PMID- 15116944 TI - Determination of potato glycoalkaloids using high-pressure liquid chromatography electrospray ionisation/mass spectrometry. AB - A method for quantifying two toxic glycoalkaloids, alpha-solanine and alpha chaconine, in potato (Solanum tuberosum) tuber tissue was developed using HPLC electrospray ionisation (ESI)/MS. Potato samples were extracted with 5% aqueous acetic acid, and the extracts were subjected directly to HPLC-ESI/MS after filtration. By determining the intensities of the protonated molecules of alpha solanine (m/z 868) and alpha-chaconine (m/z 852) using selected ion monitoring (positive ion mode), a sensitive assay was attained with detection limits of 38 and 14 ppb for the two glycoalkaloids, respectively. The high sensitivity and selectivity of MS detection effectively reduced the time of analysis thus enabling a high throughput assay of glycoalkaloids in potato tubers. PMID- 15116945 TI - Steroids and triterpenes from Eleocharis acutangula and E. sellowiana (Cyperaceae). AB - From the hexane extract of the underground parts of Eleocharis acutangula (Roxb.) Schult., lup-20(29)-ene-3beta,16beta-diol and a mixture of campesterol, stigmasterol and sitosterol were isolated. The hexane extracts of aerial and underground parts of E. sellowiana Kunth furnished two new substances, namely neohop-13(18)-en-3alpha-ol and stigmast-22-en-3beta,6beta,9alpha-triol, together with a mixture of steroids, betulinic acid, stigmast-4-en-6beta-ol-3-one and fern 9(11)-en-3alpha-ol. The molecular structures were determined by spectral analysis (1D- and 2D-NMR experiments and MS) and comparison with literature data. PMID- 15116947 TI - Childhood growth hormone deficiency (GHD) and idiopathic short stature (ISS): how far can a consensus go? PMID- 15116948 TI - Management of Turner's syndrome. PMID- 15116949 TI - How effective and safe is growth hormone therapy in children born small for gestational age? PMID- 15116950 TI - Transition from paediatric to adult endocrine care in patients with childhood onset growth hormone deficiency. PMID- 15116951 TI - An anti E. coli O157:H7 antibody-immobilized microcantilever for the detection of Escherichia coli (E. coli). AB - A silicon microcantilever sensor was developed for the detection of Escherichia coli O157:H7. The microcantilever was modified by anti-E. coli O157:H7 antibodies on the silicon surface of the cantilever. When the aquaria E. coli O157:H7 positive sample is injected into the fluid cell where the microcantilever is held, the microcantilever bends upon the recognition of the E. coli O157:H7 antigen by the antibodies on the surface of the microcantilever. A negative control sample that does not contain E. coli O157:H7 antigen did not cause any bending of the microcantilever. The detection limit of the sensor was 1 x 10(6) cfu/mL when the assay time was < 2 h. PMID- 15116952 TI - Porphyrin hybrid complex ([2Fe-2S]2TPPS) as a catalyst for determination of hydrogen peroxide. AB - A new promising mimetic enzyme, a [2Fe-2S] cluster-porphyrin hybrid complex ([2Fe 2S]2TPPS), has been synthesized and applied to the determination of hydrogen peroxide. Under the optimum condition, the calibration graph has a linear range of 8.0 x 10(-8) - 1.0 x 10(6) mol/l H2O2 with a detection limit (3sigma, N = 9) of 5.3 x 10(-9) mol/l. PMID- 15116953 TI - Detection of H2O2 released from TiO2 photocatalyst to air. AB - H2O2 generated and released from TiO2 photocatalysts to the gas phase was detected. A flow-through cell packed with TiO2-coated glass beads was irradiated with UV light, and the gas flowing out of the cell was flushed through a collecting solution containing 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulforic acid) diammonium salt (ABTS) and peroxidase. Oxidative coloration of ABTS was observed only in the absence of catalase, indicating the presence of H2O2 in the gas. The quantum yield of the H2O2 generation was estimated to be > 1 x 10(-7). The detected amount of H2O2 decreased as the TiO2 thickness decreased. H2O2 was not detected when dry air or nitrogen was used. PMID- 15116954 TI - The Human Genome Project: the role of analytical chemists. AB - The Human Genome Project (HGP) is the most ambitious and important effort in the history of biology. It has provided a complete genetic blueprint for human life, and will provide important insights into human health and development. HGP involves a huge amount of data that is stored on computers all over the world. More than just vast amounts of DNA sequences, the project is about developing sets of integrated maps that involve genetic, physical, and sequence data. The data can be sorted, annotated and organized in many different ways using different types of database software, different analysis algorithms and different forms of interfaces. The genomic sequences of the human and the substantial portions of the mouse genome are expected to be finished by 2005. Analytical chemists took the opportunity, addressing the problem of achieving a high throughput with good sensitivity. This paper discusses how analytical chemists saved the Human Genome Project or at least gave it a helping hand. PMID- 15116955 TI - Third-generation biosensors based on the direct electron transfer of proteins. AB - Recent progress in third-generation electrochemical biosensors based on the direct electron transfer of proteins is reviewed. The development of three generations of electrochemical biosensors is also simply addressed. Special attention is paid to protein-film voltammetry, which is a powerful way to obtain the direct electron transfer of proteins. Research activities on various kinds of biosensors are discussed according to the proteins (enzymes) used in the specific work. PMID- 15116956 TI - Screening of endocrine disrupting chemicals using a surface plasmon resonance sensor. AB - Because concern over endocrine disrupting reactions caused by chemicals to humans and animals is growing, a rapid and reliable screening assay for endocrine disrupting chemicals is required. We have developed an in vitro screening assay based on a hormone receptor mechanism using a surface plasmon resonance (SPR) sensor. The interaction between an estrogen receptor alpha (ER) and an estrogen response element (ERE) is monitored in real time, when ER is injected over the SPR sensor chip on which a DNA fragment containing ERE is immobilized. In the presence of a chemical with estrogenic activity, the ER-ERE interaction is enhanced and the kinetic parameters are altered. We have validated the assay in terms of its specificity, dose dependency, optimal reaction conditions and reproducibility. It has been shown that the assay is very reliable as a rapid and quantitative screening method to judge the estrogenic activities of chemicals. PMID- 15116957 TI - Isotopic analysis of Fe in human red blood cells by multiple collector-ICP-mass spectrometry. AB - Precise 56Fe/54Fe and 57Fe/54Fe isotopic ratios on human red blood cell (RBC) samples have been measured using multiple collector-ICP-mass spectrometry (MC ICPMS). The mass spectrometric interferences on Fe isotopes (e.g., 56ArO+ and 57ArOH+) were successfully minimized by a dry plasma condition achieved by a desolvating nebulizer sample-introduction technique. In order to eliminate possible variations in the measured isotopic ratios due to non-mass spectrometric interferences, Fe was separated from remaining organic compounds and major co existing elements using an ion chromatographic technique. The resulting precisions of the 56Fe/54Fe and 57Fe/54Fe ratio measurements were 0.12 per thousand and 0.20 per thousand, respectively, which were high enough to detect the isotopic variation of Fe in nature. For an interlaboratory comparison, all of the Fe isotopic ratio data were normalized by the ratios for the IRMM-014 international isotopic standard. A series of 12 RBC samples were collected from one person through monthly-based sampling over a period of one year. These were analyzed to test possible seasonal changes in the 56Fe/54Fe and 57Fe/54Fe ratios. Moreover, in order to test possible variations in the 56Fe/54Fe and 57Fe/54Fe ratios among different people, RBC samples were collected from five volunteers (four males and one female). The 56Fe/54Fe and 57Fe/54Fe ratios for a series of 12 RBC samples collected over a one-year period show 3.06 per thousand and 4.51 per thousand lower than the values of IRMM-014, and no significant seasonal change could be found in the ratios. The lack in seasonal changes in the Fe isotopic ratios could be explained by a small contribution of the daily net intake of Fe (1 - 2 mg/day) onto the total amount of Fe in the human body (2 - 4 g). The 56Fe/54Fe and 57Fe/54Fe ratios for RBC samples collected from four male samples did not vary measurably, whereas the Fe isotopic ratios for a female RBC were 0.3 per thousand/amu heavier than the mean value of four male samples. This difference in Fe isotopes among the individuals can be the result of a difference in uptake efficiency of the Fe through a dietary process from the digestive tract. The data obtained here demonstrate that the isotopic ratios of trace metals can provide new information about metabolic efficiencies of the metallic elements. PMID- 15116958 TI - Estimation of kinetics parameters for the adsorption of human serum albumin onto hydroxyapatite-modified silver electrodes by piezoelectric quartz crystal impedance analysis. AB - The adsorption of human serum albumin onto hydroxyapatite-modified silver electrodes has been in situ investigated by utilizing the piezoelectric quartz crystal impedance technique. The changes of equivalent circuit parameters were used to interpret the adsorption process. A kinetic model of two consecutive steps was derived to describe the process and compared with a first-order kinetic model by using residual analysis. The experimental data of frequency shift fitted to the model and kinetics parameters, k1, k2, psi1, psi2 and qr, were obtained. All fitted results were in reasonable agreement with the corresponding experimental results. Two adsorption constants (7.19 kJ mol(-1) and 22.89 kJ mol( 1)) were calculated according to the Arrhenius formula. PMID- 15116959 TI - Aluminum facilitation of the iron-mediated oxidation of DOPA to melanin. AB - Aluminum, a trivalent cation unable to undergo redox reactions, is shown to faciliate iron-initiated DOPA oxidation in the melanin pathway under acidic condition of pH 5.5, which is a favored medium for aluminum facilitation of iron induced lipid peroxidation. In the process of oxidation of DOPA to melanin in the presence of the metal ions, Fe3+ and H2O2 oxidize DOPA to dopachrome (DC), then Al3+ catalyzes the conversion of DC to 5,6-dihydroxyindole (DHI) and finally Fe3+ oxidizes DHI to indole-5,6-quinone (IQ), which polymerizes immediately to melanochrome and melanin. The reactions involve the intermediate complexes of metal ions and DOPA or its derivative. The present results indicate that aluminum can enhance the oxidative stress on iron-mediated DOPA oxidation in melanin pathway under acidic condition through the cooperation of iron and aluminum ions. PMID- 15116960 TI - Direct electrochemistry of xanthine oxidase at a gold electrode modified with single-wall carbon nanotubes. AB - The direct electrochemistry of xanthine oxidase (XOD) was accomplished at a gold electrode modified with single-wall carbon nanotubes (SWNTs). A pair of well defined redox peaks was obtained for XOD with the reduction peak potential at 0.478 V and a peak potential separation of 28 mV at pH 7.0. Both FT-IR spectra and the dependence of the reduction peak current on the scan rate revealed that XOD adsorbed onto the SWNT surfaces. The redox wave corresponds to the redox center of the flavin adenine dinucleotide (FAD) of the XOD adsorbate. Compared to other types of carbonaceous electrode materials, the electron transfer rate of XOD redox reaction was greatly enhanced at the SWNT-modified electrode. The peak potential was shown to be pH dependent. Spectral methods verified that the attachment of XOD onto SWNTs does not perturb the XOD conformations drastically. PMID- 15116961 TI - Adsorptive stripping voltammetric determination of azithromycin at a glassy carbon electrode modified by electrochemical oxidation. AB - The adsorptive and electrochemical behaviors of azithromycin were investigated on a glassy carbon electrode that was electrochemically treated by anodic oxidation at +1.8 V, following potential cycling in the potential range from -0.8 to +1.0 V. The resulting electrode showed good activity to improve the electrochemical response of the drug. An adsorptive stripping voltammetric method for the determination of azithromycin at an electrochemically activated glassy carbon electrode has been developed. Azithromycin was accumulated in phosphate buffer, pH 6, at a potential of +0.3 V (vs. Ag/AgCl electrode) for a certain time, and then determined by differential pulse voltammetry. The oxidative peak current at +0.82 V, at a scan rate of 20 mV s(-1), was a linear function of the concentration in the ranges of 0.25 - 2 microg mL(-1) and 1 - 10 microg mL(-1) using a 240 or 60 s(-1) preconcentration time, respectively. Application of the method to the determination of azithromycin in pharmaceuticals resulted in an acceptable deviation from the stated concentration. The preconcentration medium exchange approach was utilized for the selective determination of the drug in spiked urine samples with satisfactory results. The peak current was linear with the drug concentration in the range of 0.5 - 3.5 microg per mL urine. The detection limit was 0.2 microg mL(-1) urine. The recovery levels of the method reached 96.3%. PMID- 15116962 TI - Sequential determination of iron(II) and iron(III) in pharmaceutical by flow injection analysis with spectrophotometric detection. AB - A flow injection procedure for the sequential spectrophotometric determination of iron(II) and iron(III) in pharmaceutical products is described. The method is based on the catalytic effect of iron(II) on the oxidation of iodide by bromate at pH = 4.0. The reaction was monitored spectrophotometrically by measuring the absorbance of produced triiodide ion at 352 nm. The activating effect for the catalysis of iron(II) was extremely exhibited in the presence of oxalate ions, while oxalate acted as a masking agent for iron(III). The iron(III) in a sample solution could be determined by passing through a Cd-Hg reductor column introduced in the FIA system to reduce iron(III) to iron(II), which allows total iron determination. Under the optimum conditions, iron(II) and iron(III) could be determined over the range of 0.05 - 5.0 and 0.10 - 5.0 microg ml(-1), respectively with a sampling rate of 17 +/- 5 h(-1). The experimental limits of detection were 0.03 and 0.04 microg ml(-1) for iron(II) and iron(III), respectively. The proposed method was successfully applied to the speciation of iron in pharmaceutical products. PMID- 15116963 TI - Potentiometric determination of dopamine in pharmaceutical preparations by crown ether-PVC membrane sensors. AB - Two simple, rapid and sensitive sensors for the assay of dopamine hydrochloride have been developed. The methods are based upon the formation of the membrane sensors 12-crown-4-phosphotungestic acid (crown ether-PTA)-dopamine and 12-crown 4-tetraphenylborate (crown ether-TPB)-dopamine as neutral carriers. The sensors were stable and showed fast potential responses of 10 s, and near-Nernstian cationic slopes of 53.3 - 56.2 mV/decade of activity between pH 2.2 - 6 for the monovalent dopamine cation over a wide range concentrations 1 x 10(-5) - 1 x 10( 1) M. The selectivity coefficients of the developed sensors indicated excellent selectivity for dopamine over a large number of organic and inorganic species and pharmaceutical excipients. The mediator o-nitrophenyloctyl ether significantly affected the lifetime of the fabricated sensors of dopamine. Satisfactory results were obtained for the determination of dopamine in dosage form by the proposed sensors with an average recovery of 99.85% for the nominal concentration. PMID- 15116964 TI - Relation between electrophoretic behavior and molecular shapes of aromatic anions. AB - The capillary electrophoretic behavior of 44 aromatic organic ions was investigated. The observed ionic radii (r(obs0)) for the aromatic organic ions were obtained from the electrophoretic mobilities of sodium tetraborate (pH 9.2), potassium tetraborate (pH 9.2), ammonium borate (pH 9.2), and trisodium phosphate (pH 11.7) buffers with zero ionic strength. The linear relationships between the r(obs0)) values and the ionic radii (r(calc)), calculated by either the AM1 or PM3 method, were determined for benzyltrialkylammonium and aromatic sulfonate ions. However, the r(obs0)) values were constant for the aromatic carboxylate ions in buffers, in spite of the different r(calc) values. This indicates that aromatic carboxylate ions, such as benzenecarboxylate, pyridinecarboxylate, naphthalenecarboxylate, and anthracenecarboxylate ions, migrate as planar ions in buffers, whereas aromatic sulfonate ions could migrate as approximately spherical ions. PMID- 15116965 TI - Optimization of nonequilibrium liquid-phase microextraction for the determination of nitrobenzenes in aqueous samples by gas chromatography-electron capture detection. AB - In the present work, a novel method for the determination of nitrobenzenes in water has been described. It is based on nonequilibrium liquid-phase microextraction and gas chromatography-electron capture detection (GC-ECD). Extraction conditions such as solvent selection, organic solvent dropsize, stirring rate, content of NaCl and extraction time were found to have significant influence on extraction efficiency. The optimized conditions were 1.5 microl toluene and 20 min extraction time at 400 rpm stirring rate without NaCl addition. The linear range was 0.1 - 50 microg l(-1) for most nitrobenzenes. The limits of detection (LODs) ranged from 0.02 microg l(-1) (for 2.6-DNT) to 0.4 microg l(-1) (for NB); and relative standard deviations (RSD) for most of the nitrobenzenes at the 10 microg l(-1) level, except for 2,6-DNT in 3 microg l(-1), were below 10%. Natural samples collected from Miyun Reservoir and tap water samples from a laboratory were successfully analyzed using the proposed method, but none of the analytes were detected. The relative recoveries of spiked water samples (at the 10 microg l(-1) level except for 2,6-DNT in 3 microg l(-1)) were from 82.6 to 118.7%. PMID- 15116966 TI - Solid-phase microextraction of phenol compounds using a fused-silica fiber coated with beta-cyclodextrin-bonded silica particles. AB - A novel solid-phase microextraction (SPME) fiber was prepared by coating an HPLC beta-cyclodextrin bonded silica stationary phase (CDS) on the surface of a fused silica fiber. The fiber was evaluated for the determination of five phenol compounds (phenol, 2,4-dimethylphenol, 4-nitrophenol, 3-chlorophenol, 4 methylphenol). Compared with commercially available polymer coatings, the CDS coating showed high sensitivity and fast velocity of mass transfer for phenol compounds because of its porous structure and a unique molecular structure of beta-cyclodextrin. In addition, the CDS coating was proved to be very stable at a relatively high temperature (up to 300 degrees C). The method was suitable for the determination of phenol compounds in aqueous samples. The determination of 4 nitrophenol in soil by microwave-assisted extraction (MAE) coupled to solid-phase microextraction was also investigated. PMID- 15116967 TI - Chiral separation of binaphthol enantiomers on molecularly imprinted polymer monolith by capillary electrochromatography. AB - A novel enantioseparational monolithic stationary phase for binaphthol based on a molecular imprinting method was introduced and evaluated in capillary electrochromatography (CEC). The monolithic stationary was prepared by the in situ copolymerization of methacrylic acid and ethylene glycol dimethacrylate in a porogenic solvent (toluene or toluene-isooctane) in the presence of an imprinting molecule, (R)-1,1'-bi-2,2'-naphthol. Such stationary phases could separate the enantiomers of binaphthol. The influence of several parameters on the column permeability was investigated. These parameters included the polymerization time, the molar ratio of the functional monomer to the imprinting molecule and the content of porogen. The influence of the polymerization condition and the electrochromatographic parameters on the enantiomer separation was also studied. Initial studies showed that a higher molecular ratio of the imprinted molecule to the functional monomer, a higher content of porogen, a higher content of acetonitrile, a higher pH, as well as the addition of Tween 20, gave a higher enantiomer selectivity. PMID- 15116968 TI - Sequential injection analysis of lead using time-based colorimetric detection and preconcentration on an anionic-exchange resin. AB - The development of a sequential injection analysis manifold for the colorimetric determination of lead in water samples is described The concentration of lead was assessed from its catalytic effect on the reaction of resazurine reduction caused by sulfide in an alkali medium. To that effect, the reaction zone was stopped at the detector, and the time interval required for the attainment of an absorbance decrease of 0.800 at the wavelength of 610 nm was estimated. Interference of other transition metals of the samples was minimized by adding potassium iodide to the sample and retaining the iodocomplexes formed in an on-line anionic resin (AGI X8). Elution was made with a 2 mol/L sodium hydroxide solution. The relationship [SIA] microg/L = 0.99 (+/- 0.11) x [ETAAS] microg/L + 0 (+/- 4) was obtained upon comparing the results given by the proposed system and by electrothermal atomization atomic absorption spectrometry (ETAAS) after the analysis of ten water samples. PMID- 15116969 TI - Preconcentration of gallium by coprecipitation with synthetic zeolites prior to determination by electrothermal atomic absorption spectrometry. AB - Synthetic zeolites were dissolved in nitric acid, and the resulting solution used as a coprecipitant for the preconcentration of trace amounts of gallium in water samples prior to determination by electrothermal atomic absorption spectrometry (ETAAS). The gallium preconcentration conditions and the ETAAS measurement conditions were optimized. Gallium was quantitatively concentrated with the zeolites coprecipitate from pH 6.0 to 8.0. The coprecipitate was easily dissolved in nitric acid, and an aliquot of the resulting solution was introduced directly into a tungsten metal furnace. The atomic absorbance of gallium in the resulting solution was measured by ETAAS. An ashing temperature of 400 degrees C and an atomizing temperature of 2600 degrees C were selected. The calibration curve was linear up to 3.0 microg of gallium and passed through the origin. The detection limit (S/N > or = 3) for gallium was 0.08 microg/100 cm3. The relative standard deviation at 1.0 microg/100 cm3 was 3.0% (n = 5). The proposed method has been successfully applied to trace gallium analysis in environmental water samples. PMID- 15116970 TI - Simplification and evaluation of a gold-deposited SPR optical fiber sensor. AB - The structure of the sensing element of a gold-deposited optical fiber sensor was simplified and quantitative analyses of various alcohols with the sensor showed improvement of the performance. The sensor uses surface plasmon resonance (SPR) at the interface of a sample solution and a thin (10 - 70 nm) gold film deposited on half of the exposed core of the optical fiber. The sensor with a film thickness of 45 nm can detect a small change of 5.6 x 10(-5) refractive index (RI) units in the refractivity. The response time is less than 0.5 min and the relative standard deviation for measurements is less than or equal to 1%. A straight line with a correlation coefficient of 0.9995 was obtained below 10%, v/v in the calibration curve for methanol solutions of benzyl alcohol. The minimum of the response curve due to the maximum excitation of SPR in the refractivity range from 1.33 to 1.44 RI units shifts to a lower refractivity as the film becomes thicker. The response curves of the sensors were calculated from SPR theoretical equations while considering of the distribution in the thickness of the deposited gold films. The improvement in the performance of the sensor is discussed. PMID- 15116971 TI - The interference effect of a mixture of magnesium, aluminium, sulfate and chloride on the atomization and vaporization of manganese in graphite furnace atomic absorption spectrometry. AB - In this study, the interference effects of Al3+, Mg2+, Cl- and SO4(2-) ions on the determination of manganese by graphite furnace atomic absorption spectrometry (GFAAS) were investigated. At first, the interferences caused by Al2(SO4)3, AlCl3, MgCl2 and MgSO4, which are the most possible major compounds for the combinations of the ions mixed, were individually considered. Then, the effects caused by mixtures containing various amounts of MgSO4 and AlCl3 were studied. If the pyrolysis temperature is below 800 degrees C, AlCl3 changes the vaporization mechanism of manganese. These interferences disappear at higher pyrolysis temperatures. At the same time, aluminum salts may cause the formation of refractory compounds between aluminum and manganese (like spinel MnAl2O4) that shift the absorption signals of manganese to higher temperatures. Magnesium sulfate, by itself, does not cause any depression of manganese signals. In fact, it acts as a modifier, preventing volatilization losses of manganese during the pyrolysis step. A conclusion was reached that detailed investigation of the interferences in a complex media is a very difficult experimental and theoretical task. To solve practical problems, one may better follow the general notions developed in GFAAS toward complex matrices. PMID- 15116972 TI - Laser defocusing effects on laser ablation inductively coupled plasma-atomic emission spectrometry: different ablation interactions between the laser and low alloy steel, Fe pellets, and a pond sediment pellet. AB - The ablation interaction between a laser and solid samples, which affects the analytical performance for laser ablation inductively coupled plasma atomic emission spectrometry (LA-ICP-AES), was studied. The emission intensities of elements observed by LA-ICP-AES (LA-ICP-AES element signal intensities) for different solid samples were measured under different laser defocusing conditions with a fixed laser output energy. It was found that the optimum laser defocusing conditions were dependent on the different solid samples with different sample characteristics, and also on the different elements with different elemental characteristics in each solid sample. A low-alloy steel, pellets containing different Fe concentrations (0 - 100% Fe pellet), and a pond sediment pellet were used as different solid samples. The variations of the LA-ICP-AES Fe signal intensities observed under different laser defocus conditions were completely different between the low-alloy steel and the pond sediment pellet. The changes in the LA-ICP-AES Fe signal intensities for 90 and 100% Fe pellets were similar to that of the low-alloy steel. However, pellets with lower Fe concentrations (less than 70%) showed different trends and the defocusing behavior became closer to that of the pond sediment pellet. The LA-ICP-AES signal intensities of other elements were also evaluated, and were compared for different solid samples and different defocusing behavior. It was observed that the changes in the LA-ICP-AES signal intensities of almost all elements in the pond sediment pellet showed a similar trend to those of Fe for different laser defocus positions; that is, the elemental fractionation for these elements in the pond sediment pellet seemed to be relatively small. On the contrary, it was found that the LA-ICP-AES Si, Ti, and Zr signal intensities for low-alloy steel showed different trends compared to those of other elements, including Fe, under different defocusing conditions; that is, the elemental fractionation observed for the low-alloy steel was larger than that of the pond sediment pellet. From these results, different ablation interactions between the laser and the different solid samples were considered, and attributed to the sample characteristics, such as the matrix, hardness, and conductivity. Elemental fractionation was attempted to be explained by using elemental characteristics, such as the melting point and ionization energy of the elements. PMID- 15116973 TI - Spectrofluorometric determination of hydrogen peroxide based on oxidative catalytic reactions of p-hydroxyphenyl derivatives with metal complexes of thiacalix[4]arenetetrasulfonate on a modified anion-exchanger. AB - The peroxidase-like catalytic activity of metal complexes of thiacalix[4]arenetetrasulfonate (TCAS[4]) on a modified anion-exchanger (Me(n+) TCAS[4]A-500; Me(n+) = H2, Fe3+, Fe2+, Mn3+, Co3+, Co2+, Cu2+, Zn2+, Ni2+) for the oxidation of p-hydroxyphenyl derivatives to produce fluorescent substances in the presence of hydrogen peroxide has been investigated. Among the Me(n+) TCAS[4]A-500 tested, Fe(3+)-TCAS[4]A-500 exhibited the highest level of catalytic activity for the oxidation of p-acetoamidophenol in a carbonate buffer solution of pH 10. The catalytic activity of Fe(3+)-TCAS[4]A-500 was then used for the spectrofluorometric determination of hydrogen peroxide. The calibration curve for the Fe(3+)-TCAS[4]A-500 method was linear over a range spanning from 0.1 to 5.0 microg of hydrogen peroxide in a 1.0 ml sample solution. PMID- 15116974 TI - Electrochemical and phosphorescent properties of new Ir(III) complexes coordinated by various bipyridine derivatives. AB - Four useful polypyridine iridium(III) complexes in the form of [IrCl2L2]+ were prepared and their spectroscopic and electrochemical properties as well as X-ray crystallography were investigated. The ligands used were L = 2,2'-bipyridine, 4,4'-dimethyl-2,2'-bipyridine, 4,4'-diphenyl-2,2'-bipyridine, 1,10 phenanthroline, 4,7-diphenyl-1,10-phenanthroline, and 2,2'-biquinoline. Synthetic methods were developed by a sequential ligand-replacement, which occurred in the reaction vessel using a microwave oven. All complexes showed that LUMOs are based on the pi-system contribution of the polypyridine ligand for [IrCl2(bpy)2]+, [IrCl2(dmbpy)2]+, [IrCl2(dpbpy)2]+, [IrCl2(phen)2]+, [IrCl2(dpphen)2]+ and [IrCl2(bqn)2]+. The HOMOs are also localized on the polypyridine ligand in the iridium complexes. It was found that [IrCl2L2]+ emits intense phosphorescence at room temperature. In particular, the use of dpbpy as ancillary ligands extends the lifetime (660 ns) of the 3(pi-pi*) excited states of Ir(III) polypyridine complexes. The complex [IrCl2(bqn)2]+ with electron acceptor substituents shows a large red-shift to 622 nm. It is noticed that iridium polypyridine complexes show intense emissions at various colors, such as yellow for [IrCl2(dmbpy)2]+ and red for [IrCl2(bqn)2]+ which can be applied to photosensitizers. The spectroscopic and electrochemical details are also reported herein. PMID- 15116975 TI - Systemic analysis of structures and contents of nitrogen-containing compounds and other non-hydrocarbons in crude oils in conjunction with chemometric resolution technique. AB - A method is described for the systemic identification and quantitative analysis of nitrogen-containing compounds and other non-hydrocarbons in crude oils. The pre-fractionation of a crude oil sample into 7 fractions was performed by di adsorption column chromatography using neutral aluminum oxide and silica gel. A subsequent high-resolution separation of individual components was achieved by using capillary column gas chromatography, and compound types were detected by a mass spectrometer. In conjunction with a chemometric method, the compounds in the fractions were further resolved or separated, which made it possible to identify some nitrogen-containing compounds and other non-hydrocarbons in crude oils. To a certain extent, this method could relieve the difficulty of classical analysis in identifying those species with very low contents or incompletely separation, particularly in the cases where authentic standards were not available for addition into the unknown samples in order to reveal what indeed existed in them. The structures and contents of 168 nitrogen-containing compounds in one crude sample and 60 non-nitrogen-containing compounds in one of non-hydrocarbon fractions of this oil sample were determined, and the addition-recovery examination of some standard compounds showed that the analytical veracity was satisfactory. PMID- 15116976 TI - Novel method for the fast preconcentration and monitoring of a ppt level of lead and copper with a modified hexagonal mesoporous silica compound and inductively coupled plasma atomic emission spectrometry. AB - A novel method for the rapid extraction and determination of a ppt level of Pb2+ and Cu2+ ions using partial silylated MCM-41 modified by a new salophen and inductively coupled plasma atomic emission spectrometry (ICP-AES) is introduced. The preconcentration factor of the method is 500, and the detection limits of Pb2+ and Cu2+ are 335 and 34 ng L(-1), respectively. The time and efficiency of extraction, the pH and flow rate, the type and minimum amount of acid for stripping of Pb2+ and Cu2+ from modified MCM-41 and the break-through volume were investigated. The maximum capacity of 4 mg of silylated MCM-41 modified by salophen used was found to be 150 +/- 4 and 117 +/- 3 microg of Pb2+ and Cu2+, respectively. PMID- 15116978 TI - Co-extraction of lanthanoid(III) with various metal ions in the chelate extraction system with acetylacetone. AB - The co-extraction phenomenon was found in a typical chelate extraction system, in which the extraction of lanthanoid ion (Ln3+) with acetylacetone (Hacac) was highly enhanced by various metal ions (M(n+)) such as Cu2+, Al3+, and Zr4+. This phenomenon was ascribed to the formation of the 1:1 adduct between Ln(acac)3 and the M(acac),, extracted into the organic phase. The co-extraction occurred more readily for La3+ than that for Lu3+, and increased in the order of Cu2+ < Al3+ < Zr4+. This work elucidated that the co-extraction due to the adduct formation is a rather common phenomenon in the chelate extraction. PMID- 15116977 TI - Improvement in reliability of probabilistic test of significant differences in GeneChip experiments. AB - A probabilistic test (FUMI theory) for GeneChip experiments has been proposed for selecting the genes which show significant differences in the gene expression levels between a single pair of treatment and control. This paper describes that the reliability of the judgment by the FUMI theory can be enhanced, when the selected genes are referred to biomolecular-functional networks of a commercial database. The genes judged as being differently expressed are grouped into a cluster in the biomolecular networks. It is also demonstrated that false positive genes have a trend in the networks to be isolated from each other, and also away from the clustered genes, since the false positive genes are randomly selected. PMID- 15116979 TI - Determination of vanadium in heavy oils by atomic absorption spectrometry using a graphite furnace coated with tungsten. AB - A simple and rapid method was established for the direct determination of vanadium in an oil sample using the tungsten-coated graphite-furnace AAS. The interference of the sulfur compound could be suppressed by choosing the ashing temperature. If the sulfur concentration in the sample is diluted to 1.0 wt% or less, the interference can be suppressed. This proposed method should make important contributions to the quality control of petroleum refineries. PMID- 15116980 TI - Oxidation of isoniazid by quinolinium dichromate in an aqueous acid medium and kinetic determination of isoniazid in pure and pharmaceutical formulations. AB - The kinetics of oxidation of isoniazid in acidic medium was studied spectrophotometrically. The reaction between QDC and isoniazid in acid medium exhibits (4:1) stoichiometry (QDC:isoniazid). The reaction showed first order kinetics in quinolinium dichromate (QDC) concentration and an order of less than unity in isoniazid (INH) and acid concentrations. The oxidation reaction proceeds via a protonated QDC species, which forms a complex with isoniazid. The latter decomposes in a slow step to give a free radical derived from isoniazid and an intermediate chromium(V), which is followed, by subsequent fast steps to give the products. The reaction constants involved in the mechanism are evaluated. Isoniazid was analyzed by kinetic methods in pure and pharmaceutical formulations. PMID- 15116982 TI - "One-thousand one... one-thousand two...": chronometric counting violates the scalar property in interval timing. AB - Weber's law applied to interval timing is called the scalar property. A hallmark of timing in the seconds-to-minutes range, the scalar property is characterized by proportionality between the standard deviation of a response distribution and the duration being timed. In this temporal reproduction study, we assessed whether the scalar property was upheld when participants chronometrically counted three visually presented durations (8, 16, and 24 sec) as compared with explicitly timing durations without counting. Accuracy for timing and accuracy for counting were similar. However, whereas timing variability showed the scalar property, counting variability did not. Counting variability across intervals was accurately modeled by summing a random variable representing an individual count. A second experiment replicated the first and demonstrated that task differences were not due to presentation order or practice effects. The distinct psychophysical properties of counting and timing behaviors argue for greater attention to participant strategies in timing studies. PMID- 15116981 TI - How a cognitive psychologist came to seek universal laws. AB - My early fascination with geometry and physics and, later, with perception and imagination inspired a hope that fundamental phenomena of psychology, like those of physics, might approximate universal laws. Ensuing research led me to the following candidates, formulated in terms of distances along shortest paths in abstract representational spaces: Generalization probability decreases exponentially and discrimination time reciprocally with distance. Time to determine the identity of shapes and, provisionally, relation between musical tones or keys increases linearly with distance. Invariance of the laws is achieved by constructing the representational spaces from psychological rather than physical data (using multidimensional scaling) and from considerations of geometry, group theory, and symmetry. Universality of the laws is suggested by their behavioral approximation in cognitively advanced species and by theoretical considerations of optimality. Just possibly, not only physics but also psychology can aspire to laws that ultimately reflect mathematical constraints, such as those of group theory and symmetry, and, so, are both universal and nonarbitrary. PMID- 15116983 TI - Geometrical haptic illusions: the role of exploration in the Muller-Lyer, vertical-horizontal, and Delboeuf illusions. AB - This article surveys studies of the occurrence, in the haptic modality, of three geometrical illusions well known in vision, and it discusses the nature of the processes underlying these haptic illusions. We argue that the apparently contradictory results found in the literature concerning them may be explained, at least partially, by the characteristics of manual exploratory movements. The Muller-Lyer illusion is present in vision and in haptics and seems to be the result of similar processes in the two modalities. The vertical-horizontal illusion also exists in vision and haptics but is due partly to similar processes (bisection) and partly to processes specific to each modality (anisotropy of the visual field and overestimation of radial vs. tangential manual exploratory movements). The Delboeuf illusion seems to occur only in vision, probably because exploration by the index finger may exclude the misleading context from tactile perception. The role of these haptic exploratory movements may explain why haptics is as sensitive as vision to certain illusions and less sensitive to others. PMID- 15116984 TI - Spatial stimulus-response compatibility and negative priming. AB - According to Kornblum's (1992) dimensional overlap model, when an incongruent response to a stimulus is required, automatic activation of the congruent response must first be inhibited. Shiu and Kornblum (1996a) provided evidence for such inhibition in an incongruent symbolic negative priming task. Reaction time was longer when a trial's correct response was the name of the stimulus from the previous trial than when it was not. We report three experiments that test this inhibition hypothesis for spatial stimuli and responses. In Experiment 1, which used a spatial mapping analogous to the symbolic mapping used by Shiu and Kornblum (1996a), a similar negative priming effect was found. However, in Experiments 2 and 3, which used mappings that were conducive to simple transformational rules, a positive priming effect was obtained. The results suggest that inhibition in response selection may depend on the complexity of the relations between the stimuli and responses. PMID- 15116985 TI - Spatial Simon effects with nonspatial responses. AB - Recent studies have shown that spatial Simon effects can be modulated by short term associations that are set up as a result of task instructions. I examined whether spatial Simon effects can also be produced by short-term associations even when the responses are unrelated to spatial position. Participants were to say "cale" or "cole" on the basis of the direction of arrows (i.e., left or right), the meaning of words (i.e., left or right), and the color of squares presented left or right of the screen center. Responses to squares were faster when the correct response was associated with the same position as the irrelevant position of the square (e.g., say "cale" to a square on the left when "cale" was assigned to the word left and the left arrow). This new type of stimulus-response compatibility effect provides the first evidence for short-term associations that involve mode-independent representations. PMID- 15116986 TI - Spatial perception and control. AB - We investigated whether the perceived vanishing point of a moving stimulus becomes more accurate as one's degree of control over the stimulus increases. Either alone or as a member of a pair, participants controlled the progression of a dot stimulus back and forth across a computer monitor. They did so via right and left buttonpresses that incremented the dot's velocity rightward and leftward, respectively. The participants in the individual condition had control of both buttons. Those in the group condition had control of only one. As the participants slowed the dot to change its direction of travel, it unexpectedly disappeared. Localizations of the vanishing point became more accurate as the participants' control over the dot increased. The data bridge a gap between accounts of localization error that rely solely on stimulus and cognitive factors, and accounts derived from research on action and spatial perception, which tend to rely on action-planning factors. PMID- 15116987 TI - A new estimation of the duration of attentional dwell time. AB - How rapidly can attention move from one object to the next? Previous studies in which the dwell time paradigm was used have estimated attentional switch times of 200-500 msec, results incompatible with the search rate estimates of 25-50 msec shown in numerous visual search studies. It has been argued that dwell times are so long in the dwell time paradigm because the attentional shifts measured are unlike those used in visual search. In the present experiment, a variation of a visual search task was used, in which serial endogenous (volitional) deployments of attention were measured directly by means of a probe reaction time task. The experiment revealed a dwell time of about 250 msec, consistent with the faster estimates from other dwell time studies. This result suggests that endogenous shifts of attention may be relatively slow and that the faster attentional shifts estimated from visual search tasks may be due to the involvement of bottom-up processes. PMID- 15116988 TI - Top-down search strategies cannot override attentional capture. AB - Bacon and Egeth (1994) have claimed that color singletons do not interfere with search for a shape singleton when, instead of using a singleton detection mode, participants are forced to use a feature search mode. Bacon and Egeth induced a feature search mode by adding different shape singletons to the display so that observers could not simply respond to uniqueness to find the target. We did exactly the same but used larger display sizes to ensure that the target and distractor singletons remained salient. The results show that under these conditions, an irrelevant color singleton interferes with search for a shape singleton. It is argued that the notion of differential search modes may be incorrect and that the results can be explained in terms of bottom-up salience signals. PMID- 15116989 TI - Search for multiple targets: evidence for memory-based control of attention. AB - There are two opposing models with regard to the function of memory in visual search: a memory-driven model and a memory-free model. Recently, Horowitz and Wolfe (2001) investigated a multiple-target search task. Participants were required to decide whether or not there were at least n targets present. They demonstrated that the reaction time x n function has a positive and accelerated curve. They argued that the memory-free model predicts this curve, whereas the memory-driven model predicts a linear function. In this study, I varied the total set sizes of a multiple-target search task and fitted the models separately for each n condition. The model fit indicated that the memory-driven model is more appropriate than the memory-free model in each n condition. These results suggest that an amnesic process does not cause the positive accelerated curve of the reaction time x n function but that it is the result of the time needed to examine each additional n item. PMID- 15116990 TI - Dissociating sources of dual-task interference using human electrophysiology. AB - In the psychological refractory period (PRP) paradigm, two unmasked targets are presented, each of which requires a speeded response. Response times to the second target (T2) are slowed when T2 is presented shortly after the first target (T1). Electrophysiological studies have previously shown that the P3 event related potential component is not delayed during T2 response slowing in the PRP paradigm, but that the lateralized readiness potential is delayed, which suggests a bottleneck on response selection operations but not on stimulus identification. Recently, researchers (Arnell & Duncan, 2002; Jolicoeur & Dell'Acqua, 1999) observed T2 response slowing in an encoding-speeded response (ESR) paradigm where T2 followed a masked T1 that required identification but not a speeded response. T2 response slowing in the ESR paradigm is often indistinguishable from that in the PRP paradigm, prompting some researchers to postulate a common processing bottleneck for the two paradigms. With the use of the ESR paradigm, we observed T2 response slowing and, in contrast to the PRP paradigm, we also observed corresponding P3 delays. The results suggest that dissociable bottlenecks underlie the dual-task costs from the two paradigms. PMID- 15116991 TI - Hemispheric performance in object-based attention. AB - The goal of the present study was to investigate whether object-based attention effects differ across the cerebral hemispheres. Previous research has suggested that object-based attention is preferentially lateralized to the left hemisphere (Egly, Driver, & Rafal, 1994; Egly, Rafal, Driver, & Starrveveld, 1994). However, work by Vecera (1994) has suggested that these previous studies may have failed to obtain a pure measure of object-based attention. The present study applied modified versions of Duncan's (1984) seminal object-based attention paradigm. Subjects were typically presented with one target object to a single visual field (one-object display), two target objects to the same visual field (two-object unilateral display), or two target objects to different visual fields (two-object bilateral display). In all three experiments, response accuracy was higher for the one-object displays than for the two-object displays. Most important, this object-based cost was especially severe when selection of two target elements was isolated to the right visual field (left hemisphere). We confirmed that this effect was specific to object-based attention in three different ways: Experiment 1 manipulated stimulus distance, as recommended by Vecera; Experiment 2 ensured that target selection was based on nonspatial attributes; and Experiment 3 used overlapping displays, as in Duncan (1984). Collectively, the data are in accord with previous conclusions that object-based attention is a specialized form of orienting subserved by lateralized cortical brain mechanisms. However, contrary to previous research, it appears that it is the right hemisphere, and not the left hemisphere, that is preferentially biased for committing object-based attention to elements in the visual environment. PMID- 15116992 TI - Is the asymmetry in young infants' categorization of humans versus nonhuman animals based on head, body, or global gestalt information? AB - Quinn and Eimas (1998) reported an asymmetry in the exclusivity of the category representations that young infants form for humans and nonhuman animals: category representations for nonhuman animal species were found to exclude humans, whereas a category representation for humans was found to include nonhuman animal species (i.e., cats, horses). The present experiment utilized the familiarization/novelty preference procedure with 3- and 4-month-olds to determine the perceptual cues (i.e., whole stimulus, head alone, body alone) that provided the basis for this asymmetry. The data revealed the asymmetry to be observable only with the whole animal stimuli and not when infants were provided with information from just the head or the body of the exemplars. The results indicate that the incorporation of nonhuman animal species into a category representation for humans is based on holistic information. PMID- 15116993 TI - The Beck effect is back, now in color: a demonstration. AB - It has been repeatedly found (e.g., Beck, 1982; Beck & Ambler, 1973) that a tilted T is detected better than an L when presented on the background of a number of upright Ts, although in isolation both are discriminated from an upright T about equally well. We demonstrate that an analogous interaction between context presence and stimulus type can be obtained with color stimuli: A light green was discriminated from a dark green better than an olive brown was, yet that occurred only in the context of a background of three dark green disks. This indicates that the effects, both original and novel, probably are not due to anything peculiar to form perception, such as line arrangement or supraelement configurations. It is suggested that the presence of collateral background stimuli probably enables the perceptual system to capitalize on relative judgment involving contrasts or deviations from a presented anchor value. PMID- 15116994 TI - Testing quantitative models of backward masking. AB - We analyzed the relationship between U-shaped and monotonic-shaped masking functions, using both computer simulations of quantitative models and experimental data Our analysis revealed that quantitative models of backward masking predict that U-shaped masking functions should appear for weak masks and monotonic masking functions should appear for strong masks. The models predict, moreover, that for a fixed target and experimental task, as the mask changes it is possible to go from U-shaped to monotonic-shaped masking functions. Significantly, the models predict that at each stimulus onset asynchrony between the target and the mask, the U-shaped function must have weaker masking than the monotonic-shaped function. Contrary to the predictions of the models, we show an experimental situation that generates masking functions that violate this prediction. PMID- 15116995 TI - Pigeons group time intervals according to their relative duration. AB - In the present research, we asked whether pigeons tended to judge time intervals not only in terms of their absolute value but also relative to a duration from which they must be discriminated (i.e., longer or shorter). Pigeons were trained on two independent temporal discriminations. In one discrimination, sample durations of 2 and 8 sec were associated with, for example, red and green hue comparisons, respectively, and in the other discrimination, sample durations of 4 and 16 sec were associated with vertical and horizontal line comparisons, respectively. If pigeons are trained on a temporal discrimination and tested with intermediate durations, the subjective midpoint typically occurs close to the geometric mean of the two trained values. The 4- and 8-sec values were selected to be the geometric mean of the two values in the other discrimination. When a 4 sec test sample was presented with the comparisons from the 2- and 8-sec discrimination, the pigeons preferred the comparison associated with the shorter sample. Similarly, when an 8-sec test sample was presented with the comparisons from the 4- and 16-sec discrimination, the pigeons preferred the comparison associated with the longer sample. Thus, a relative grouping effect was found. That is, durations that should have produced indifferent choice were influenced by their relative durations (shorter than or longer than the alternative) during training. PMID- 15116996 TI - Decomposing serial learning: what is missing from the learning curve? AB - Our current understanding of serial learning relies on the form of the learning curve and on changes in the serial position curve over repeated study-test trials (Ward, 1937). The averaging of data that produces these functions obscures the detailed history of memory for individual items over the course of study-test trials. Extending Tulving's (1964) analysis of free recall learning, we present a new analysis of serial learning that tracks the acquisition and forgetting of item and order information at the level of individual items. Applying this analysis to two large data sets on serial list learning allows us to discern among hypotheses that are indistinguishable solely on the basis of the learning curve. PMID- 15116997 TI - Retrieval-induced forgetting occurs in tests of item recognition. AB - Using the retrieval-practice paradigm (Anderson, R. A. Bjork, & E. L. Bjork, 1994), we tested whether or not retrieval-induced forgetting could be found in item recognition tests. In Experiment 1, retrieval practice on items from semantic categories depressed recognition of nonpracticed items from the same categories. Similar results were found in Experiment 2 in a more stringent source test for practiced, nonpracticed, and new items. These results conceptually replicate those of previous retrieval-induced forgetting studies done with cued recall (e.g., Anderson et al., 1994). Our findings are inconsistent with the hypothesis that item-specific cues during retrieval will eliminate retrieval interference in the retrieval-practice paradigm (Butler, Williams, Zacks, & Maki, 2001). We discuss our results in relation to other retrieval interference and inhibition effects in recall and recognition. PMID- 15116998 TI - Self-evaluation as a moderating factor of strategy change in directed forgetting benefits. AB - In list method directed forgetting, instructing people to forget a studied word list usually results in better recall for a newly studied list. Sahakyan and Delaney (2003) have suggested that these benefits are due to a change in encoding strategy that occurs between the study of the first list and the study of the second list. To investigate what might mediate such strategy change decisions, in two experiments we induced both forget and remember participants to evaluate their memory performance on the two lists. In Experiment 1, they were asked to explicitly recall the items from the first list before studying the second list. In Experiment 2, after the study of the first list, the participants provided a rapid aggregate judgment of learning. Evaluation eliminated the differences between the forget and remember groups for the second list in both experiments, because the remember group achieved recall levels comparable to those for the forget group. The role of performance evaluation in mediating directed forgetting benefits is discussed. PMID- 15116999 TI - False recognition without intentional learning. AB - Asked to memorize a list of semantically related words, participants often falsely recall or recognize a highly related semantic associate that has not been presented (the critical lure). Does this false memory phenomenon depend on intentional word reading and learning? In Experiment 1, participants performed a color identification task on distractor words from typical false memory lists. In Experiment 2, participants read the same words. In both experiments, the primary task was followed by a surprise recognition test for actually presented and unpresented words, including the critical lures. False alarms to critical lures were robust and quite equivalent across the two experiments. These results are consistent with an activation/monitoring account of false memory, in which processing of semantic associates can evoke false memories even when that processing is incidental. PMID- 15117000 TI - Effects of perceptual modality on verbatim and gist memory. AB - In two experiments, predictions of the fuzzy-trace theory of memory were tested. Perceptual information may play a role in retrieval and recognition processes for verbatim, but not for gist, memory. Perceptual modality effects were assessed in the present study by presenting three-sentence stories (e.g., The bird is in the cage. The cage is over the table. The bird is yellow) and then testing recognition of probes that varied on three dimensions: (1) semantic accuracy (true vs. false), (2) wording (all original words vs. one novel word included), and (3) sentence type (premise vs. inference). In Experiment 1, study modality (auditory vs. visual) was manipulated, and in Experiment 2, both study and test modalities were manipulated. Despite replicating a number of findings consistent with fuzzy-trace theory (e.g., instruction and probe type effects), the results of both experiments failed to support the idea that perceptual information plays a role in performance on verbatim memory tests. PMID- 15117001 TI - The role of spurious feature familiarity in recognition memory. AB - In two experiments, we investigated the role of perceptual information in spurious recognition judgments. Participants viewed lists of words in various unusual fonts. The frequency with which each font was presented was manipulated at study: Each font was presented with 1 or 12 different words in Experiment 1 and with 1 or 20 words in Experiment 2. Although the participants were instructed in a word recognition test to judge only on the basis of the word, regardless of font, there were significantly more false alarms for new words seen in a previously presented font than for new words presented in a novel (not seen at study) font in Experiment 1. In Experiment 2, the participants were significantly more likely to make a false alarm to a new word seen in a font that had been used to present 20 words during study than to a font that had been used to present only 1 word during study. The data show a mirror effect, in which words tested in low-frequency fonts produced more hits and fewer false alarms than did words tested in high-frequency fonts. These results show that irrelevant perceptual information plays a role in recognition judgments by providing spurious sources of familiarity and, thus, provide evidence that perceptual information is represented and processed in the same way as semantic information. PMID- 15117002 TI - Body-based senses enhance knowledge of directions in large-scale environments. AB - Previous research has shown that inertial cues resulting from passive transport through a large environment do not necessarily facilitate acquiring knowledge about its layout. Here we examine whether the additional body-based cues that result from active movement facilitate the acquisition of spatial knowledge. Three groups of participants learned locations along an 840-m route. One group walked the route during learning, allowing access to body-based cues (i.e., vestibular, proprioceptive, and efferent information). Another group learned by sitting in the laboratory, watching videos made from the first group. A third group watched a specially made video that minimized potentially confusing head-on trunk rotations of the viewpoint. All groups were tested on their knowledge of directions in the environment as well as on its configural properties. Having access to body-based information reduced pointing error by a small but significant amount. Regardless of the sensory information available during learning, participants exhibited strikingly common biases. PMID- 15117003 TI - Sensorimotor simulations underlie conceptual representations: modality-specific effects of prior activation. AB - According to the perceptual symbols theory (Barsalou, 1999), sensorimotor simulations underlie the representation of concepts. Simulations are componential in the sense that they vary with the context in which the concept is presented. In the present study, we investigated whether representations are affected by recent experiences with a concept. Concept names (e.g., APPLE) were presented twice in a property verification task with a different property on each occasion. The two properties were either from the same perceptual modality (e.g., green, shiny) or from different modalities (e.g., tart, shiny). All stimuli were words. There was a lag of several intervening trials between the first and second presentation. Verification times and error rates for the second presentation of the concept were higher if the properties were from different modalities than if they were from the same modality. PMID- 15117004 TI - The self-choice effect from a multiple-cue perspective. AB - The self-choice effect refers to the fact that self-chosen items are remembered better than experimenter-assigned items (Takahashi, 1991). The present study investigated the hypothesis that (a) response choice involves relational processing as activation of both target and context items, and (b) such activated context items are effective as potential retrieval cues for recall of target items. In the experiment, participants chose (choice condition) or were assigned (force condition) a target to remember for each trial. Prior to free recall of the target items, context words, related new words, or unrelated words were presented in a recognition task as potential retrieval cues. The results of a subsequent free recall test indicated that the incidental cues were more effective in the choice condition than in the force condition. Also, recognition resulted in a greater rate of successfully recognized context words at the cost of increasing falsely recognized related new words in the choice condition in comparison with the force condition. These results indicated that response choice activates context items at encoding, which operate as potential retrieval cues for recall of target items. Such cuing mechanisms operative in the self-choice effect are consistent with the multiple-cue theory proposed by Soraci et al. (1994; see also Soraci et al., 1999) for generative processing. PMID- 15117005 TI - The effects of accountability on bias in physician decision making: going from bad to worse. AB - Members of the Iowa Academy of Family Physicians participated in a survey study in which they were asked to make hypothetical decisions in either high- or low conflict treatment conditions. In the low-conflict treatment condition the options were a common medication with a referral or a referral only. In the high conflict treatment condition the same two options were given plus another attractive medication. In addition, we manipulated accountability by asking half of the participants to provide a written defense of their treatment options, which they would then agree to discuss at a later time. The results showed that physicians in the high-conflict condition chose the referral-only option significantly more than the physicians in the low-conflict condition, thus violating the normative rule of regularity. Moreover, that pattern was significantly amplified for physicians who were held accountable for their treatment decisions. These findings replicate and extend previous research and are discussed within the framework of reason-based choice. PMID- 15117006 TI - Randomness and inductions from streaks: "gambler's fallacy" versus "hot hand". AB - Sometimes people believe that a run of similar independent events will be broken (belief in the gambler's fallacy) but, other times, that such a run will continue (belief in the hot hand). Both of these opposite inductions have been explained as being due to belief in a law of small numbers. We argue that one factor that distinguishes these phenomena is people's beliefs about the randomness of the underlying process generating the events. We gave participants information about a streak of events but varied the scenarios in such a way that the mechanism generating the events should vary in how random the participants would judge it to be. A manipulation check confirmed our assumptions about the scenarios. We found that with less random scenarios, the participants were more likely to continue a streak. PMID- 15117007 TI - Protected values: no omission bias and no framing effects. AB - Previous studies have suggested that people holding protected values (PVs) show a bias against harmful acts, as opposed to harmful omissions (omission bias). In the present study, we (1) investigated the relationship between PVs and acts versus omissions in risky choices, using a paradigm in which act and omission biases were presented in a symmetrical manner, and (2) examined whether people holding PVs respond differently to framing manipulations. Participants were given environmental scenarios and were asked to make choices between actions and omissions. Both the framing of the outcomes (positive vs. negative) and the outcome certainty (risky vs. certain) were manipulated. In contrast to previous studies, PVs were linked to preferences for acts, rather than for omissions. PVs were more likely to be associated with moral obligations to act than with moral prohibitions against action. Strikingly, people with strong PVs were immune to framing; participants with few PVs showed robust framing effects. PMID- 15117008 TI - AIC model selection using Akaike weights. AB - The Akaike information criterion (AIC; Akaike, 1973) is a popular method for comparing the adequacy of multiple, possibly nonnested models. Current practice in cognitive psychology is to accept a single model on the basis of only the "raw" AIC values, making it difficult to unambiguously interpret the observed AIC differences in terms of a continuous measure such as probability. Here we demonstrate that AIC values can be easily transformed to so-called Akaike weights (e.g., Akaike, 1978, 1979; Bozdogan, 1987; Burnham & Anderson, 2002), which can be directly interpreted as conditional probabilities for each model. We show by example how these Akaike weights can greatly facilitate the interpretation of the results of AIC model comparison procedures. PMID- 15117009 TI - Treatments for patients exposed to bioterrorism agents. PMID- 15117010 TI - Alternatives to ear syringing for removal of earwax. PMID- 15117011 TI - Metabolic syndrome: early identification and a proactive approach are warranted. PMID- 15117012 TI - Abnormal uterine bleeding. AB - Abnormal uterine bleeding is a common presenting symptom in the family practice setting. In women of childbearing age, a methodical history, physical examination, and laboratory evaluation may enable the physician to rule out causes such as pregnancy and pregnancy-related disorders, medications, iatrogenic causes, systemic conditions, and obvious genital tract pathology. Dysfunctional uterine bleeding (anovulatory or ovulatory) is diagnosed by exclusion of these causes. In women of childbearing age who are at high risk for endometrial cancer, the initial evaluation includes endometrial biopsy; saline-infusion sonohysterography or diagnostic hysteroscopy is performed if initial studies are inconclusive or the bleeding continues. Women of childbearing age who are at low risk for endometrial cancer may be assessed initially by transvaginal ultrasonography. Postmenopausal women with abnormal uterine bleeding should be offered dilatation and curettage; if they are poor candidates for general anesthesia or decline dilatation and curettage, they may be offered transvaginal ultrasonography or saline-infusion sonohysterography with directed endometrial biopsy. Medical management of anovulatory dysfunctional uterine bleeding may include oral contraceptive pills or cyclic progestins. Menorrhagia is managed most effectively with nonsteroidal anti-inflammatory drugs or the levonorgestrel intrauterine contraceptive device. Surgical management may include hysterectomy or less invasive, uterus-sparing procedures. PMID- 15117013 TI - Information from your family doctor. Abnormal uterine bleeding. PMID- 15117014 TI - Identifying the vector of Lyme disease. AB - Lyme disease is the most common vector-borne illness in the United States. It is caused by the spirochete Borrelia burgdorferi, which is transmitted by the deer tick. Deer ticks have a four-stage life cycle (egg, larva, nymph, and adult), and nymphal ticks transmit B. burgdorferi to humans more frequently than adult ticks. Transmission of this spirochete typically requires a minimum of 24 to 48 hours of tick attachment. Early stages of Lyme disease are characterized by a hallmark rash, erythema migrans. The overall risk of acquiring Lyme disease is low in a person who has a deer tick bite. If erythema migrans develops at the site of the bite, treatment may include doxycycline in persons who are at least eight years of age. Administration of amoxicillin is appropriate for pregnant women or children younger than eight years. For those who are allergic to these medications, cefuroxime axetil may be used. PMID- 15117015 TI - Hand and wrist injuries: Part I. Nonemergent evaluation. AB - Diagnosis of upper extremity injuries depends on knowledge of basic anatomy and biomechanics of the hand and wrist. The wrist is composed of two rows of carpal bones. Flexor and extensor tendons cross the wrist to allow function of the hand and digits. The ulnar, median, and radial nerves provide innervation of the hand and wrist. A systematic primary and secondary examination of the hand and wrist includes assessment of active and passive range of motion of the wrist and digits, and dynamic stability testing. The most commonly fractured bone of the wrist is the scaphoid, and the most common ligamentous instability involves the scaphoid and lunate. PMID- 15117016 TI - Hand and wrist injuries: Part II. Emergent evaluation. AB - Primary care physicians must be able to recognize wrist and hand injuries that require immediate attention. A complete history and physical examination, including assessment of distal limb function, are essential. Hemorrhage control is necessary in patients with vessel lacerations and amputations. Amputations require an understanding of the indications and contraindications in the management of the amputated limb. High-pressure injection injuries and compartment syndromes require a high index of suspicion for early recognition. Infectious entities include "fight bite," open fractures, purulent tenosynovitis, animal bites, and retained foreign bodies. Tendon disruptions should be recognized early to optimize management. PMID- 15117018 TI - Information from your family doctor. Taking steps to prevent type 2 diabetes. PMID- 15117017 TI - Impaired glucose tolerance and impaired fasting glucose. AB - Impaired glucose tolerance and impaired fasting glucose form an intermediate stage in the natural history of diabetes mellitus. From 10 to 15 percent of adults in the United States have one of these conditions. Impaired glucose tolerance is defined as two-hour glucose levels of 140 to 199 mg per dL (7.8 to 11.0 mmol) on the 75-g oral glucose tolerance test, and impaired fasting glucose is defined as glucose levels of 100 to 125 mg per dL (5.6 to 6.9 mmol per L) in fasting patients. These glucose levels are above normal but below the level that is diagnostic for diabetes. Patients with impaired glucose tolerance or impaired fasting glucose have a significant risk of developing diabetes and thus are an important target group for primary prevention. Risk factors for diabetes include family history of diabetes, body mass index greater than 25 kg per m2, sedentary lifestyle, hypertension, dyslipidemia, history of gestational diabetes or large for-gestational-age infant, and polycystic ovary syndrome. Blacks, Latin Americans, Native Americans, and Asian-Pacific Islanders also are at increased risk for diabetes. Patients at higher risk should be screened with a fasting plasma glucose level. When the diagnosis of impaired glucose tolerance or impaired fasting glucose is made, physicians should counsel patients to lose 5 to 7 percent of their body weight and engage in moderate physical activity for at least 150 minutes per week. Drug therapy with metformin or acarbose has been shown to delay or prevent the onset of diabetes. However, medications are not as effective as lifestyle changes, and it is not known if treatment with these drugs is cost effective in the management of impaired glucose tolerance. PMID- 15117019 TI - Screening for obesity in adults: recommendations and rationale. PMID- 15117020 TI - Preoperative evaluation for noncardiac surgery. PMID- 15117022 TI - Photo quiz. Acute shortness of breath after transfusion. PMID- 15117021 TI - Teriparatide (Forteo) for osteoporosis. PMID- 15117023 TI - ACS guidelines for early detection of cancer. PMID- 15117024 TI - Selective stimulation of the canine hypoglossal nerve using a multi-contact cuff electrode. AB - Electrical activation of the tongue protrusor muscle has been demonstrated as an effective technique for alleviating upper airway (UAW) obstructions and is considered a potential treatment for obstructive sleep apnea (OSA). Recent studies, however, have shown marked improvements in UAW patency by coactivating the tongue protrudor and retractor muscles. As such, selective stimulation of the hypoglossal nerve (XII) using a single implantable device presents an attractive approach for treating OSA. In order to demonstrate the feasibility of such a device, the maximum achievable stimulation selectivity of the Flat Interface Nerve Electrode (FINE) was investigated. The XII nerve of beagles was stimulated with an acutely implanted FINE, while the corresponding neural and muscular responses were recorded and analyzed. The overall performance of the FINE, as depicted by the average of the maximum target-specific selectivity values, S(i), confirmed that high degrees of selectivity can be achieved at both the fascicular and muscular levels: 0.93 +/- 0.03 (n = 5) and 0.88 +/- 0.03 (n = 4), respectively. The results of this study demonstrate the feasibility of the FINE for selective stimulation of the XII nerve branches and the innervated tongue muscles. PMID- 15117025 TI - A computational tensegrity model predicts dynamic rheological behaviors in living cells. AB - Rheological properties of living cells play a key role in the control of cell shape, growth, movement, and contractility, yet little is known about how these properties are governed. Past approaches to understanding cell mechanics focused on the contributions of membranes, the viscous cytoplasm, and the individual filamentous biopolymers that are found within the cytoskeleton. In contrast, recent work has revealed that the dynamic mechanical behavior of cells depends on generic system properties, rather than on a single molecular property of the cell. In this paper, we show that a mathematical model of cell mechanics that depicts the intracellular cytoskeleton as a tensegrity structure composed of a prestressed network of interconnected microfilaments, microtubules, and intermediate filaments, and that has previously explained static cellular properties, also can predict fundamental dynamic behaviors of living cells. PMID- 15117026 TI - Cell membrane fluidity changes and membrane undulations observed using a laser scattering technique. AB - Local transversal micromotions of cell membranes have been reported. These may provide the physical basis for changes in membrane fluidity. In this study, tissue scattering properties are used to measure the magnitude of transversal micromotions under varying stimuli. Laser light is directed at the cell surface at a low incident angle of 12 degrees so that reflected or refracted light does not enter the microscope objective. Scattered light showed strong low-frequency intensity fluctuations with random-walk behavior. Fluctuations were quantified by the median coefficient of variation over the entire observed cell area. Incubation of the cells with protein crosslinkers (paraformaldehyde) and metabolic suppressors (sodium azide) suppressed these fluctuations by 62 and 44%, respectively. The application of hypoosmotic media caused an increase of fluctuation magnitude by 23% (p < 0.005). Agents that increase membrane fluidity (2% ethanol and xenon) increased fluctuation magnitude by 15 and 31%, respectively (p < 0.05). Cessation of the ethanol and xenon exposure led to partial recovery of the fluctuation magnitude, which was nonsignificant for ethanol. This study shows a strong link between membrane fluidity and transversal membrane undulations and provides an important step in the understanding of the mechanosensing function of the cell membrane. PMID- 15117027 TI - The effects of a shear flow on the uptake of LDL and acetylated LDL by an EC monoculture and an EC-SMC coculture. AB - To elucidate the mechanisms of localized genesis and development of atherosclerosis and anastomotic intimal hyperplasia in man, a coculture of bovine aortic endothelial cells (ECs) and smooth muscle cells (SMCs) was prepared, and the effects of a shear flow on the uptake of lipoproteins by the cells was studied by incubating the EC-SMC coculture as well as an EC monoculture with a culture medium containing either DiI-LDL or DiI-Ac-LDL and subjecting to a laminar shear flow. It was found that in both the presence and absence of a shear flow that imposed the ECs an area mean shear stress of 13.3 dynes/cm2, the uptake of LDL by an EC-SMC coculture was much greater than that by an EC monoculture, whereas that of Ac-LDL was almost the same. The uptake of LDL by an EC monoculture increased slightly by being exposed to a shear flow, whereas that by an EC-SMC coculture did not. In contrast to this, the uptake of Ac-LDL by both an EC monoculture and an EC-SMC coculture decreased drastically by a shear flow, suggesting that the action of a shear flow on the uptake of Ac-LDL by vascular cells is very different from that of LDL. PMID- 15117029 TI - Effects of constant static pressure on the biological properties of porcine aortic valve leaflets. AB - An understanding of how mechanical forces impact cells within valve leaflets would greatly benefit the development of a tissue-engineered heart valve. In this study, the effect of constant ambient pressure on the biological properties of heart valve leaflets was evaluated using a custom-designed pressure system. Native porcine aortic valve leaflets were exposed to static pressures of 100, 140, or 170 mmHg for 48 h. Collagen synthesis, DNA synthesis, sulfated glycoaminoglycan (sGAG) synthesis, alpha-SMC actin expression, and extracellular matrix (ECM) structure were examined. Results showed that elevated pressure caused an increase in collagen synthesis. This increase was not statistically significant at 100 mmHg, but at 140 mmHg and 170 mmHg collagen synthesis increased by 37.5 and 90%, respectively. No significant difference in DNA or sGAG synthesis was observed at elevated pressures, with the exception that DNA synthesis at 100 mmHg decreased. A notable decline in alpha-SMC actin was observed over the course of the experiments although no significant difference was observed between the pressure and control groups. It was concluded that elevated pressure caused a proportional increase in collagen synthesis of porcine aortic valve leaflets, but was unable to preserve alpha-SMC actin immunoreactive cells. PMID- 15117028 TI - Alteration in fluid mechanics in porcine femoral arteries with atheroma development. AB - Although the relationship between the local wall shear stresses (WSS) and atheroma development has been previously studied, the effect of variable regional wall distensibility with early atherosclerotic lesions and its effect on local hemodynamics have not been well investigated. We induced early atherosclerotic lesion development in one femoral artery in a Yucatan miniswine model with the contralateral artery serving as control. Eight weeks following atheroma formation, hemodynamic and intravascular ultrasound image data were obtained. Using the measured regional arterial distension as the moving boundary condition, unsteady laminar incompressible computational analysis was performed on the reconstructed arterial segments. With the development of early atherosclerotic lesions, there was increased wall distensibility and an increase in the computed normalized wall shear stress amplitude (NWSA). Alterations in the local fluid mechanics and mass transport at these sites may need to be considered in our understanding of the continued growth of these lesions. PMID- 15117030 TI - Stress relaxation preconditioning of porcine aortic valves. AB - In uniaxial tensile testing, load preconditioning is used to generate repeatable load/elongation curves and set a "reference state" for subsequent tensile tests. We have observed however, that for porcine aortic valve (PAV) tissues, preconditioning does not lead to repeatable stress relaxation curves. We thus investigated possible experimental protocols that could be used to generate repeatable load/elongation and stress relaxation curves. To quantify repeatability of stress relaxation, we compared normalized loads at the same time points from repeated stress relaxation curves and computed a repeatability ratio. We found that PAV specimens can generate repeatable stress relaxation curves (repeatability ratio >0.95) if they are subjected to at least five cycles of repeated load preconditioning and stress relaxation. We also found that a single cycle of loading/unloading prior to each stress relaxation phase is sufficient to generate repeatable stress relaxation curves. Stress relaxation preconditioning is therefore required to generate repeatable load/elongation and stress relaxation curves. It is expected that such curves will generate more accurate material constants for the characterization and modeling of PAV mechanics. PMID- 15117031 TI - Novel noncontact catheter system for endocardial electrical and anatomical imaging. AB - The study objective was to integrate noncontact mapping and intracardiac echocardiography (ICE) in a single catheter system that enables both electrical and anatomical imaging of the endocardium. We developed a catheter system on the basis of a 9-F sheath that carried a coaxial 64-electrode lumen-probe on the outside and a central ICE catheter (9 F, 9 MHz) on the inside. The sheath was placed in the right atrium (RA) of 3 dogs, and in the left ventricle (LV) of 3 other dogs. To construct cardiac anatomy, the ICE catheter was pulled back over several beats inside the sheath starting from the tip and two-dimensional tomographic images were continuously acquired. To recover endocardial electrograms, the probe was advanced over the sheath and single-beat noncontact electrograms were simultaneously recorded. Endocardial contact electrodes were placed at select sites for validation as well as for pacing. Three-dimensional electrical-anatomical images reconstructed during sinus and paced rhythms correctly associated RA and LV activation sequences with underlying endocardial anatomy (overall activation error = 3.4 +/- 3.2 ms; overall spatial error = 8.0 +/- 3.5 mm). Therefore, accurate fusion of electrical imaging with anatomical imaging during catheterization is feasible. Integrating single-beat noncontact mapping with ICE provides detailed, three-dimensional electrical-anatomical images of the endocardium, which may facilitate management of arrhythmias. PMID- 15117032 TI - Modeling RBC and neutrophil distribution through an anatomically based pulmonary capillary network. AB - An anatomically based finite element model of the human pulmonary microcirculation has been created and applied to simulating regional variations in blood flow. A geometric mesh of the capillary network over the surface of a single alveolar sac is created using a Voronoi meshing technique. A pressure-flow relationship that describes blood cell transit is implemented in the network. Regional flow is investigated by imposing gravity-dependent transpulmonary and transmural boundary conditions. Comparisons of red and white blood cell transit times in the upper, mid, and lower lung showed physiologically consistent trends of a decreasing average transit time and an increased homogeneity of transit time distributions as a result of increasing average capillary diameter and flow down the height of a vertical lung. The model was found to reproduce experimentally consistent trends in red blood cell transit times and relative blood flows with respect to lung height. This model enables flow properties and cell transit time behavior in the pulmonary microcirculation under varying conditions, for example in different "zones" of the lung, to be explored. PMID- 15117033 TI - Servo-controlled pneumatic pressure oscillator for respiratory impedance measurements and high-frequency ventilation. AB - The ability to provide forced oscillatory excitation of the respiratory system can be useful in mechanical impedance measurements as well as high frequency ventilation (HFV). Experimental systems currently used for generating forced oscillations are limited in their ability to provide high amplitude flows or maintain the respiratory system at a constant mean pressure during excitation. This paper presents the design and implementation of a pneumatic pressure oscillator based on a proportional solenoid valve. The device is capable of providing forced oscillatory excitations to the respiratory system over a bandwidth suitable for mechanical impedance measurements and HVF. It delivers high amplitude flows (> 1.4 l/s) and utilizes a servo-control mechanism to maintain a load at a fixed mean pressure during simultaneous oscillation. Under open-loop conditions, the device exhibited a static hysteresis of approximately 7%, while its dynamic magnitude and phase responses were flat out to 10 Hz. Broad band measurement of total harmonic distortion was approximately 19%. Under closed loop conditions, the oscillator was able to maintain a mechanical test load at both positive and negative mean pressures during oscillatory excitations from 0.1 to 10.0 Hz. Impedance of the test load agreed closely with theoretical predictions. We conclude that this servo-controlled oscillator can be a useful tool for respiratory impedance measurements as well as HFV. PMID- 15117034 TI - Modeling water vapor and heat transfer in the normal and the intubated airways. AB - Intubation of the artificially ventilated patient with an endotracheal tube bypasses the usual conditioning regions of the nose and mouth. In this situation any deficit in heat or moisture in the air is compensated for by evaporation and thermal transfer from the pulmonary airway walls. To study the dynamics of heat and water transport in the intubated airway, a coupled system of nonlinear equations is solved in airway models with symmetric geometry and anatomically based geometry. Radial distribution of heat, water vapor, and velocity in the airway are described by power-law equations. Solution of the time-dependent system of equations yields dynamic airstream and mucosal temperatures and air humidity. Comparison of model results with two independent experimental studies in the normal and intubated airway shows a close correlation over a wide range of minute ventilation. Using the anatomically based model a range of spatially distributed temperature paths is demonstrated, which highlights the model's ability to predict thermal behavior in airway regions currently inaccessible to measurement. Accurate representation of conducting airway geometry is shown to be necessary for simulating mouth-breathing at rates between 15 and 100 l x min(-1), but symmetric geometry is adequate for the low minute ventilation and warm inspired air conditions that are generally supplied to the intubated patient. PMID- 15117035 TI - Hypothermic machine preservation in liver transplantation revisited: concepts and criteria in the new millennium. AB - To overcome the present shortage of liver donors by expansion of the existing donor pool and possibly lengthening of the storage time, hypothermic machine perfusion of the liver as a dynamic preservation method is revisited. The three most important aspects are defined to be the type of preservation solution, the characteristics of perfusion dynamics, and the oxygen supply. Reviewing hypothermic liver machine perfusion experiments, the University of Wisconsin machine preservation solution is the solution most used. It is also found that nothing conclusive can be said about the optimal perfusion characteristics, since either perfusion pressure or perfusion flow is reported. The best estimation is perfusion of the liver in a physiological manner, i.e. pulsatile arterial perfusion and continuous portal venous perfusion. The applied pressures could be chosen to be somewhat lower than physiological pressures to prevent possible endothelial cell damage. Oxygen supply is necessary to achieve optimal preservation of the liver. The minimal amount of partial oxygen pressure required is inversely related to the normalized flow. Incorporating these features in a system based on existing standard surgical and organ sharing procedures and which is able to work stand-alone for 24 h, weighing less than 23 kg, could successfully implement this technique into every day clinical practise. PMID- 15117036 TI - Vascular loop compressing facial nerve in hemifacial spasm: demonstrated by 3D phase contrast magnetic resonance angiography in 101 patients. AB - BACKGROUND: Vascular compression of the facial nerve is deemed to be the common cause of hemifacial spasm producing emphatic transmission. Although facial nucleus supersensitivity is more accepted as the main cause of hemifacial spasm. PURPOSE: To determine the vascular loop compression of the facial nerve in patients with hemifacial spasm by 3D-phase contrast (PC) magnatic resonance imaging (MRI). MATERIAL AND METHOD: A retrospective study of 101 patients with hemifacial spasm who went MRI and magnetic resonance angiography (MRA) of the brain was done. The magnitude images of the 3D-PC MRA was evaluated in axial and oblique coronal reconstruction planes blindly from symptomatic information. RESULTS: Among 101 patients, 53 affected the left side, 48 patients were right sided and none had bilateral involvement. Vascular loop compressing on the symptomatic side was found in 61 (60.4%) patients. For the asymptomatic side, there were 14 (13.86%) with vascular loop contact. Five patients (4.9%) had bilateral vascular compression. The proportion of vascular contact of the symptomatic and asymptomatic side was significantly different (with p < 0.001). The offending vessels were vertebral artery (32, 52.46%), posterior inferior cerebellar artery (7, 6.93%), anterior inferior cerebellar artery (6, 5.94%) and artery of uncertain origin (16, 26.23%). CONCLUSION: The study implied the usefulness of this simple technique to demonstrate the neurovascular contact of the facial nerve. PMID- 15117038 TI - Red blood cell vesicles in thalassemia. AB - Vesicles are part of the red blood cells membrane which can be found in a small number in normal apoptotic process and increased in some diseases. In the present study, the authors measured the percentage of red blood cell vesicles in healthy subjects (n = 7), patients with alpha-thalassemia or Hemoglobin (Hb) H disease (n = 7), beta-thal/Hb E with nonsplenectomized (n = 5) and splenectomized (n = 7) before and after induction heated at 48.6 degrees C by using flow cytometry. It was found that the percentage of vesicles in every group were not statistically significantly different (p > 0.05) between pre and post incubation at 5 min. The percentage of vesicles of healthy subjects, beta-thal/Hb E nonsplenectomized patients and splenectomized patients were highest when induced by heating for 60 min. For patients with Hb H disease, the percentage of vesicles was maximum at 30 min when compared with healthy subjects, beta-thal/Hb E nonsplenectomized patients and splenectomized patients, respectively. In the present study, the authors report the significant increase of the percentage of vesicles in Hb H disease, beta-thal/Hb E nonsplenectomized and splenectomized after induction by heat when compared with healthy subjects. These findings may support the different pathology of the red blood cells found in alpha- and beta-thalassemia. PMID- 15117037 TI - Stent placement compared with balloon angioplasty for obstructed coronary artery disease in Thai elderly patients: initial result and 6 months follow-up. AB - Percutaneous Old Balloon Angioplasty (POBA) is accepted worldwide for the treatment of obstructive coronary artery disease because this technique is safe, and quick and the patient may return to work earlier than with bypass surgery (particularly elderly patients). But the major problem with POBA is restenosis which occurs between 20-40 per cent. Stent placement has been reported to reduce the restenosis rate to 10-20 per cent. OBJECTIVE: The purpose of this study was to compare the effects of stent placement with those of balloon angioplasty on clinical and angiographic outcomes in elderly Thai patients with obstructive coronary artery disease. METHOD: The study was a randomized controlled trial. The sample size was 45 lesions in 42 patients who were assigned into 2 groups; 23 in the balloon angioplasty group and 22 in the stent placement group (Crown Stent). Clinical information and coronary angiography were recorded and performed at the time of the index procedure and six months later. RESULTS: There was 100 per cent procedural success in 22 lesions treated in the stent group and 82.6 per cent in the 23 lesions treated in the balloon angioplasty group. Patients in the stent group had a lesser degree of stenosis immediately after the procedure (8.78 +/- 8.63 vs 30.92 +/- 9.01%, p < 0.001) and a greater minimal luminal diameter (MLD) (3.04 +/- 0.44 vs 2.15 +/- 0.33 mm, p < 0.001). There were no major complications in either group during the procedure or during their hospital stay. These were not maintained at the six months follow-up. (26.88 +/- 16.23 vs 33.82 +/- 14.63 mm, p = 0.19, 2.28 +/- 0.67 vs 2.01 +/- 0.51 mm, p = 0.17) for the degree of stenosis and the MLD respectively. The restenosis rate, which was the primary endpoint of the study, was 4.5 per cent in the stent group and 21 per cent in the balloon angioplasty group (p = 0.10). CONCLUSIONS: Stenting in Thai elderly ischemic patients has a higher procedural success rate when compared with balloon angioplasty. The restenosis rate of stenting is also lower than that of balloon angioplasty but did not reach statistical significance. However, both techniques had no major complications either during the procedure or in-hospital. PMID- 15117039 TI - Results of radiotherapy in non round cell spinal metastasis. AB - Spinal metastases are commonly encountered by physicians in a variety of clinical fields. There are some controversies in choice of treatment between surgery and radiotherapy. This report is a study of the outcomes of radiotherapy for metastatic nonround cell tumors of the spine. Medical records and films of 31 patients who were treated with radiotherapy at Songklanakarind Hospital were retrospectively reviewed. The most common primary tumors were prostate and breast. One patient had spinal metastases from malignant serous cystadenoma of the fallopian tube of which no previous report has been published. This patient had excellent results after radiotherapy. Back and neck pain were the primary symptoms of the patients, while motor or sensory deficits (or both) were found in 58 per cent of the cases. Seven patients had neurological recovery and 18 patients had pain relief after radiotherapy. Cause of compression is the only factor effecting the result from univariate and multivariate analysis. Spinal cord compressed by a tumor had a better recovery than those which were compressed by a bony fragment or intervertebral disc. The authors concluded that radiotherapy remains a good treatment for patient with non round cell spinal metastasis. Cause of spinal cord compression is the only factor predicting the result of treatment. PMID- 15117040 TI - Comparison of topical lomefloxacin 0.3 per cent versus topical ciprofloxacin 0.3 per cent for the treatment of presumed bacterial corneal ulcers. AB - PURPOSE: To compare the efficacy and safety of topical lomefloxacin 0.3 per cent with topical ciprofloxacin 0.3 per cent for treating mildly severe suspected bacterial corneal ulcers. METHOD: This prospective, randomized, double-masked controlled clinical trial was conducted on 41 patients (41 eyes) with suspected bacterial corneal ulcers who were randomized into 2 groups: 23 patients were in the lomefloxacin group and 18 patients in the ciprofloxacin group. All of these corneal ulcers were scraped for gram's stain, KOH preparation and microbiologic cultures before starting treatment. The clinical success rate, the time to cure, the rates of treatment failures, ocular signs and symptoms and the adverse effects of the study medication were evaluated. RESULTS: Topical lomefloxacin is equivalent clinically and statistically to topical ciprofloxacin. No statistically significant treatment differences were found between lomefloxacin (100%) and ciprofloxacin (100%) in terms of success rate. Similarly, no differences were noted in the time to cure (p > 0.05), the treatment failure, or the resolution of the clinical signs and symptoms (p > 0.05). The adverse effects of lomefloxacin were superficial punctate keratitis (26.1%) and irritation (8.7%), whereas those of ciprofloxacin were superficial punctate keratitis (22.2%), white precipitate (11.1%) and irritation (11.1%). However, no statistically significant differences of these adverse effects were found between the two groups (p > 0.05). CONCLUSION: Lomefloxacin ophthalmic solution (0.3%) is equivalent clinically and statistically to ciprofloxacin ophthalmic solution (0.3%) for the treatment of mildly severe presumed bacterial corneal ulcers without statistically significant differences in the adverse effects and discomfort. PMID- 15117041 TI - The effects of potassium and magnesium supplementations on urinary risk factors of renal stone patients. AB - The effects of potassium and magnesium supplementation on urinary risk factors for renal stone disease were studied in 61 renal stone patients. The subjects were divided into four groups and supplemented for a period of one month with potassium chloride (KCl, Group 1), potassium sodium citrate (K Na citrate, Group 2), magnesium glycine (Mg glycine, Group 3) and potassium magnesium citrate (K Mg citrate, Group 4) with a daily dose of 42 mEq potassium, 21 mEq magnesium or sodium and 63 mEq citrate, accordingly. The results showed that serum potassium and magnesium of all four groups normalized after the supplementation. Though urinary potassium significantly increased in all three groups supplemented with elemental potassium containing solutions [i.e. KCl (p < 0.001), K Na citrate (p < 0.001) and K Mg citrate (p < 0.001)] only K Na citrate and K Mg citrate, caused a significant increase in urinary pH and citrate but decrease in calcium. Supplementation with Mg glycine in Group 3 although caused a significant increase in urinary magnesium, its effects on urinary pH, citrate and calcium, however, were similar to KCl, in that they caused a significant decrease in urinary pH without any change in urinary citrate or calcium. Supplementation with K Mg citrate in Group 4 seems to have given the best results, as far as lowering stone risk factors in that it caused an increase in urinary pH, potassium and citrate and decreased calcium excretions similar to K Na citrate in Group 2. In addition, K Mg citrate also caused the enrichment of urine with magnesium, another inhibitor of calcium-containing stones. Although the four supplements had no effect on urinary saturation of calcium oxalate salt, their effects on the saturations of brushite (CaHPO4 x 2H2O), octacalcium phosphate (Ca8H2 (PO4)6 x 5H2O) and uric acid were clearly associated with changes in urinary pH. Therefore, in Group 1 and 3, subjects having a decrease in urinary pH, also experienced a significant increase in uric acid saturation. Though the saturation of brushite and octacalcium phosphate in Group 2 and 4 and the sodium acid urate in Group 2 were significantly increased, these urinary risk factors could be overcome, however, by the concomitant increase in urinary citrate. The present results demonstrate that for those stone vulnerable subjects having a high risk of potassium and magnesium depletion, to obtain the best therapeutic results, they should be provided supplementations of both potassium and magnesium together and also in the forms that would result in the delivery of an alkali loading effect. PMID- 15117042 TI - The effect of ephedrine on the onset time of rocuronium in Thai patients. AB - The aim of this study was to determine the effect of ephedrine on the onset time of rocuronium. The study population was 60 ASA physical status 1 and 2 patients, aged 15-60 years scheduled for elective surgery under general anesthesia at Ramathibodi Hospital. The patients were randomly assigned into 2 groups. Group I (ephedrine group), ephedrine 70 microg/kg was given 1 minute before induction and group II (control group), saline was given instead of ephedrine and midazolam 7.5 mg was given orally 30-60 minutes before the induction. Anesthesia was induced with fentanyl 1 microg/kg and sodium thiopentone 3-5 mg/kg. The patient was intubated with 0.9 mg/kg of rocuronium. The intubation time (from rocuronium administration to the time of intubation) was predetermined by the Dixon's up and down method (with 5 seconds as a step size) for each patient and started at 60 seconds for the first patient in each group. The intubation time in the ephedrine group (39.41 +/- 4.64 seconds) was significantly different from the control group (59.17 +/- 9.00 seconds); p-value < 0.01. The hemodynamics were similar in both groups. CONCLUSION: Intravenous ephedrine shortened the onset time of rocuronium with no significant adverse hemodynamic effects. As an alternative to suxamethonium for rapid intubation, the authors recommend the use of ephedrine 70 microg/kg at one minute before induction followed by 0.9 mg/kg of rocuronium intravenously in healthy patients. The intubation could be achieved at 40 seconds after the administration. PMID- 15117043 TI - Associated risk factors of human papillomavirus cervical infection among human immunodificiency virus-seropositive women at Siriraj Hospital. AB - This cross sectional type sub-study was established to assess the potential risk factor associated with human papillomavirus (HPV) cervical infection in Human immunodeficiency virus (HIV)-seropositive women. The series of 178 HIV seropositive women was enrolled in the Department of Obstetrics & Gynecology, Siriraj Hospital. Demographic, obstetrical and behavioral risk factors were interviewed. Laboratory results were recorded. Clinical gynecologic examination was performed including Pap smear. The patients were assigned into two groups, HPV and non-HPV group. The comparison of the potential risk factors between the groups was calculated statistically. It was found that the prevalence of HPV infection was reported in 17 patients (9.6%). HIV-seropositive women, who were infected with HPV, had a significantly more probability to have a single partner in their lifetime than those who were not infected. It could be that HPV cervical infection and HIV-seropositive women share common potential risk factors, as well as, the recognition of sexual intercourse as the important route of HPV transmission. PMID- 15117044 TI - Nasal polyps: the relationship to allergy, sinonasal infection and histopathological type. AB - OBJECTIVE: To evaluate the relationship of nasal polyps to allergy, sinonasal infection and histopathological type by examining the prevalences of these factors among nasal polyps patients. STUDY DESIGN: Prospective descriptive study. MATERIAL AND METHOD: A total of 73 patients were enrolled between October 1st, 1999 and August 31st, 2002 at the Allergy and Rhinology Clinic, Faculty of Medicine, Songklanagarind Hospital. The medical history was recorded. Allergy skin prick test, nasal endoscopy with biopsy and plain film paranasal sinus were performed. Positive allergy skin test was defined by at least 1 aeroallergen with a wheal size > or = 3 mm greater than the negative control. Rhinosinusitis was diagnosed by clinical symptoms, positive nasal endoscopy and/or positive plain film paranasal sinus. Histopathological investigation was classified as eosinophil- or neutrophil-dominated inflammation. RESULTS: 68.5 per cent of patients with nasal polyps had a positive allergy skin test, 67.1 per cent had rhinosinusitis. Eosinophil-dominated inflammation was presented in 69.9 per cent and neutrophil-dominated inflammation in 30.1 per cent, respectively. Within each histopathological type, 62.7 per cent of patients with eosinophil-dominated inflammation and 81.8 per cent of patients with neutrophil-dominated inflammation had a positive allergy skin test. There was no statistically significant difference in prevalence of positive allergy skin test between eosinophil- and neutrophil-dominated inflammations (p = 0.107). 60.8 per cent of patients with eosinophil-dominated inflammation and 81.8 per cent of patients with neutrophil dominated inflammation had rhinosinusitis. There was no statistically significant difference in prevalence of rhinosinusitis between eosinophil- and neutrophil dominated inflammations (p = 0.079). CONCLUSION: Nasal polyps had association with positive allergy skin test (68.5%), rhinosinusitis (67.1%) and eosinophil dominated inflammation (69.9%). There were no statistically significant differences in prevalence of positive allergy skin test and rhinosinusitis between eosinophil- and neutrophil-dominated inflammations (p = 0.107 and p = 0.079, respectively). PMID- 15117046 TI - A comparison of nutritional screening tools in the prediction of post-operative infectious and wound complications in the elderly patients undergoing abdominal operations. AB - OBJECTIVE: To compare four nutritional screening tools--the Short Form Mini Nutritional Assessment (MNA-SF), the Nutrition Risk Classification (NRC), the Malnutrition Screening Tool (MST), and the Nutrition Risk Score (NRS)--in the prediction of post-operative infectious and wound complications in elderly patients undergoing abdominal surgery. PATIENTS AND METHOD: During the nine-month period from April 2002 to December 2002 nutritional screening was performed on 190 patients aged 60 years or over who underwent major abdominal surgery. Each patient was classified as either at risk or not at risk of malnutrition. This classification, for each screening tool, was tested for association with the occurrence of post-operative infectious and wound complications. Each screening tool was calculated and compared under the area of Receiver Operating Characteristic (ROC) curve. RESULT AND CONCLUSION: The NRC classification was the best prediction for the occurrence of post-operative infectious and wound complications in elderly surgical patients. PMID- 15117045 TI - The preemptive analgesic effect of celecoxib for day-case diagnostic laparoscopy. AB - In a randomized trial, the preemptive analgesic effect of celecoxib in 110 infertile women undergoing day-case diagnostic laparoscopy was studied at King Chulalongkorn Memorial Hospital, Bangkok, Thailand. The patients randomly received either 200 mg celecoxib or placebo orally 2 hours before diagnostic laparoscopy. The post-operative shoulder pain and wound pain were self assessed and recorded, using Visual Analogue Scores (VAS) at 1, 2, 4, 12, and 24 hours. Total post-operative analgesic requirements were recorded at 24 hours. The mean Visual Analogue Scores (VAS) of shoulder pain in celecoxib group was statistically lower than those of the placebo group (p = 0.04). Nevertheless, the mean VAS of wound pain and the total post-operative analgesic requirements were not significantly different. It was concluded that the preemptive celecoxib in day-case diagnostic laparoscopy might have the advantage of decreasing post laparoscopic shoulder pain. PMID- 15117047 TI - Accuracy in diagnosis of acute appendicitis by comparing serum C-reactive protein measurements, Alvarado score and clinical impression of surgeons. AB - OBJECTIVE: To compare the accuracy of a surgeon's clinical diagnosis of acute appendicitis with Alvarado's predictive model and C-reactive protein (CRP) measurements. METHOD: The records of 231 adult patients between 14-75 years admitted to the hospital with suspected appendicitis from August 1999 to November 2001 were studied prospectively. Serum CRP measurements (217 patients) and Alvarado scores (231 patients) were performed before operations but were not taken into account prior to the decision to perform a laparotomy to compare the surgeon's clinical diagnosis. RESULTS: Based on the surgeon's clinical diagnosis, 193 patients underwent surgery, and 38 patients were observed. Histopathologic findings found acute appendicitis, confirming the surgeon's clinical impression, in 178 patients (positive predictive value = 92%) and normal appendix in 15 patients. Of the observed patients, 8 subsequently underwent operation for appendicitis (negative predictive value = 79%). Compared with the surgeon's clinical diagnosis (sensitivity 96% and specificity 67%), diagnosis based on an Alvarado score of > or = 7 had a lower sensitivity (79%) and that based on CRP of > 10 mg/l a much lower sensitivity (62%) and lower specificity (56%). Overall accuracy of these three diagnostic modalities were 90 per cent, 72 per cent and 61 per cent, respectively. However, median serum CRP value increased from 5 mg/l (range 3-188 mg/l) in patients with normal appendix, to 14 mg/l (range 3-222 mg/l) in patients with non-perforated appendicitis and 65 mg/l (range 3-213 mg/l) in patients with perforated or gangrenous appendicitis. CONCLUSION: The clinical assessment in diagnosing appendicitis by an experienced surgeon remains reliable and superior to either Alvarado score or CRP measurement. Nevertheless, Alvarado score and serum CRP measurements may be of value to the inexperienced surgeon, and a high Alvarado score and serum CRP should not be ignored. PMID- 15117048 TI - Transplant renal artery stenosis in Thailand. AB - Transplant renal artery stenosis (TRAS) is one of the common vascular complications post kidney transplantation. A retrospective study of TRAS among transplant recipients at a single transplant center in Thailand was performed from February 1986 to December 2002. Among 750 cases, 16 cases (2.1%) of TRAS were identified. Twelve cases (3.3%) were from cadaveric donors and four cases (1%) were from living-related donors (p-value = 0.034). Most cases presented with progressive deterioration of kidney graft with or without refractory hypertension. Doppler ultrasonography was used for initial screening followed by renal angiography. Fifteen cases were treated by Percutaneous Transluminal Angioplasty (PTA) with a 73 per cent success rate. Five cases underwent surgical revascularization with an 80 per cent success rate. Two cases (13%) of successful PTA showed recurrent stenosis with 46 months follow-up which were successfully treated by repeated PTA with stents. PMID- 15117049 TI - Conservative regimen for chronic critical limb ischemia. AB - OBJECTIVE: The objective of this study was to determine the effectiveness of the treatment of chronic critical limb ischemia by conservative regimen. METHOD: Data for all patients who underwent a conservative regimen at a single institution from January 1997 to December 2001 were entered into the registry. Conservative regimen consisted of cilostazol (Pletaal) 200 mg/day, a vegetarian diet, had completely stopped smoking and had progressive walking training. RESULTS: A total of 53 patients (59 limbs) with chronic critical limb ischemia were treated with a conservative regimen. The conservative regimen failed in 19 limbs (32.2%). In the failed limbs, infrainguinal bypass was performed on 8 limbs, aortoiliac endarterectomy was performed on 1 limb and 6 had primary amputation. The other four limbs were treated conservatively until death because of very poor cardiac function. Post-operatively, 2 grafts had thrombosis and led to amputation. CONCLUSION: These early results appear to be promising with 67.8 per cent limb saving. This conservative regimen may be appropriately performed in selected chronic critical limb ischemia, especially those who presented with clinical severe claudication, rest pain or nonhealed ulcer. Cilostazol administration may play a positive role in gangrenous limbs. PMID- 15117050 TI - Evaluation of safety and efficacy of transdermal therapeutic system-fentanyl in adult patients with gynecological cancer-related pain. AB - OBJECTIVES: To assess the analgesic safety and efficacy of Transdermal Therapeutic System (TTS)-fentanyl in the treatment of chronic gynecological cancer-related pain. BACKGROUND: TTS-fentanyl is a Transdermal Therapeutic System, which contains a rate-limiting membrane that provides constant release of fentanyl. TTS-fentanyl can be properly used to control pain. Therefore, this trial was designed to establish the analgesic efficacy and safety of TTS-fentanyl in the treatment of chronic gynecological cancer-related pain. MATERIAL AND METHOD: Thirty patients were recruited into the study. This open study was comprised of two phases. Phase 1: an oral morphine stabilization phase where eligible patients, who took other opioids and/or analgesic drugs, were entered into the stabilization phase and should be converted to oral morphine according to the conversion chart. The patients were then titrated to a stable oral morphine dose. Phase 2: an open TTS-fentanyl treatment phase where the daily dose of oral morphine was switched to TTS-fentanyl according to the conversion chart. The efficacy parameters of pain score were assessed by visual analogue scale (VAS) and global assessments. The safety was evaluated by monitoring the patient's clinical conditions and adverse events. RESULTS: TTS-fentanyl was generally well tolerated. Only one patient was dropped out from the study due to lacking enrollment in the stabilization phase. The most frequent adverse events were mild nausea or vomiting (46%) and constipation (33%). The median pain VAS during TTS-fentanyl treatment was decreased from 8 to 3 and global assessments at the end of the treatment were better than at the start of the treatment. CONCLUSION: The results suggest that TTS-fentanyl is safe and effective in managing chronic gynecological cancer-related pain. PMID- 15117051 TI - Correlation of uterine cervical length measurement from transabdominal, transperineal and transvaginal ultrasonography. AB - OBJECTIVE: To investigate the feasibility of transperineal ultrasonography for uterine cervical assessment by determining the correlation of uterine cervical length measurement from transabdominal, transperineal and transvaginal ultrasonography and comparing discomfort arising from each technique. MATERIAL AND METHOD: Fifty pregnant women of 37 weeks' gestation or later who gave consent participated in this research. They had no exclusion criteria, which were listed as the following: preterm premature rupture of membranes, previous cervical surgery, undiagnosed vaginal bleeding, and true labor pain. They all underwent transabdominal (3.5-MHz curvilinear transducer), transperineal (3.5-MHz curvilinear transducer) and transvaginal ultrasonography (7.5-MHz curvilinear endovaginal transducer). The uterine cervical length was measured from the straight line between the external and internal os. If either of the external or internal os was not clearly demonstrated, the authors would justify the cervical length as non-measurable. Measurement in each technique was performed twice and the mean cervical length was used for data analysis. Discomfort arising from each technique was evaluated by visual analog scale. RESULTS: Uterine cervical length was measurable in 23 (46%), 49 (98%) and in all cases by transabdominal, transperineal and transvaginal ultrasonography respectively. In the transabdominal technique, no significant differences in woman's age, weight, body mass index and parity were observed between measurable and non-measurable cases. Significant correlation was demonstrated between transperineal and transvaginal ultrasound (r = 0.73, p < 0.01). A significantly higher discomfort score was demonstrated in transvaginal ultrasonography, but no significant difference in discomfort score was found between transabdominal and transperineal ultrasonography. CONCLUSION: Transperineal ultrasonography is feasible for acceptable uterine cervical visualization with only slight discomfort to the patients. PMID- 15117052 TI - Acute lupus hemophagocytic syndrome: report of a case and review of the literature. AB - The authors reported a case of systemic lupus erythematosus (SLE) with an unusual presentation. The patient presented with acute febrile illness along with progressive pancytopenia related to increasing hemophagocytic activity of histiocytes in the bone marrow. Concomitant polyarthritis, myositis, nephritis, high titer of antinuclear factor (1:2,560) and positive test for anti-DNA antibody made him fit the diagnostic criteria of SLE. No definite evidence of associated infections was confirmed by bacteriologic, serologic and viral studies. He did not respond to empiric antibiotic therapy but dramatically responded to corticosteroid treatment. Therefore, diagnosis of acute lupus hemophagocytic syndrome was made. The clinical presentation, laboratory diagnosis, and management of the patient are discussed and the literature was reviewed and presented. PMID- 15117053 TI - Examination on ethics 2: Friday August 22, 2003. PMID- 15117054 TI - The dog in the fight. PMID- 15117055 TI - Double-masked, placebo-controlled, randomized trial of lutein and antioxidant supplementation in the intervention of atrophic age-related macular degeneration: the Veterans LAST study (Lutein Antioxidant Supplementation Trial). AB - BACKGROUND: Age-related macular degeneration (ARMD) is the leading cause of vision loss in aging Westem societies. The objective of the lutein antioxidant supplementation trial (LAST) is to determine whether nutritional supplementation with lutein or lutein together with antioxidants, vitamins, and minerals, improves visual function and symptoms in atrophic ARMD. METHODS: The study was a prospective, 12-month, randomized, double-masked, placebo-controlled trial conducted at an urban midwestern Veterans Administration Hospital from August 1999 to May 2001. Ninety patients with atrophic ARMD were referred by ophthalmologists at two Chicago-area veterans medical facilities. Patients in Group 1 received lutein 10 mg (L); in Group 2, a lutein 10 mg/antioxidants/vitamins and minerals broad spectrum supplementation formula (L/A); and in Group 3, a maltodextrin placebo (P) over 12 months. RESULTS: In Groups 1 L and 2 L/A, mean eye macular pigment optical density increased approximately 0.09 log units from baseline, Snellen equivalent visual acuity improved 5.4 letters for Group 1 L and 3.5 letters for Group 2 L/A, and contrast sensitivity improved. There was a net subjective improvement in Amsler grid in Group 1 L. VFO-14 questionnaires conceming subjective glare recovery were nearly significant at 4 months for Group 2 L/A. Patients who received the placebo (Group 3) had no significant changes in any of the measured findings. CONCLUSION: In this study, visual function is improved with lutein alone or lutein together with other nutrients. Further studies are needed with more patients, of both genders, and for longer periods of time to assess long-term effects of lutein or lutein together with a broad spectrum of antioxidants, vitamins, and minerals in the treatment of atrophic age-related macular degeneration. PMID- 15117056 TI - Comparisons of dynamic retinoscopy measurements with a print card, a video display terminal, and a PRIO System Tester as test targets. AB - BACKGROUND: Dynamic retinoscopy is a common clinical test for assessment of visual function in patients with symptoms related to computer use. It has been suggested that performing dynamic retinoscopy with a fixation target resembling a computer screen may be valuable in the examination of computer users. However, such targets differ in luminance and other characteristics from the printed paper test cards typically used for dynamic retinoscopy. METHOD: To address the effect of varying test targets on dynamic retinoscopy results, this study compared lag of accommodation results from dynamic retinoscopy with three different fixation targets: (a) a printed paper target, (b) a video display terminal, and (c) the PRIO vision testing device, which is designed to simulate a computer screen. MEM (monocular estimate method) dynamic retinoscopy was performed with the print target and with the PRIO device. RESULTS: The difference between results with the different test conditions were generally small, with a statistically significant 0.32 D lower lag found on MEM retinoscopy with the PRIO target IMEM-PRIO) than on MEM retinoscopy with a print target (MEM-PRINT). Mean lag measurements on MEM with a print target and on Nott retinoscopy with the video display terminal (NOTT VDT) were only 0.04 D different. The 95% limits of agreement for the difference between MEM-PRINT and MEM-PRIO (+/- 0.6 D) was similar to the 95% limits of agreement for the interexaminer reliability of the two procedures (+/- 0.6 D for MEM-PRINT and +/- 0.7 D for MEM-PRIO). CONCLUSION: There was a small but statistically significant difference in lag of accommodation measured with MEM dynamic retinoscopy on the PRIO testing device when compared to a print card. PMID- 15117057 TI - Full-time occlusion compared to part-time occlusion for the treatment of amblyopia. AB - BACKGROUND: Occlusion of the better-seeing eye as a method of treatment for amblyopia has long been a standard of care. A difference exists between practitioners on using either full-time occlusion or part-time occlusion. METHODS: A retrospective review of pediatric patients (ages 3 to 7 years) with amblyopia was performed. The patients were prescribed either full-time occlusion or part-time occlusion for the primary treatment of their amblyopia. Exclusions were made for patients with an organic or structural component of their amblyopia. Twenty-one patients were included in the full-time occlusion group and 24 patients were included in the part-time occlusion group. RESULTS: Seventy-six percent of patients who completed full-time occlusion achieved 20/40 acuity or better and 67% achieved 20/30 or better, with an average treatment time of six weeks. Fifty-eight percent of patients who completed part-time occlusion achieved 20/40 acuity or better and only 46% achieved 20/30 or better, with an average treatment time of 26 weeks. For patients with 20/80 or worse amblyopia, 82% achieved 20/40 or better with full-time occlusion. Only 40% of part-time occluders with 20/80 or worse achieved 20/40. Occlusion amblyopia did not develop in any patient. CONCLUSION: A higher percentage of amblyopic patients treated with full-time occlusion achieved 20/30 acuity in the amblyopic eye over a shorter duration of treatment. PMID- 15117059 TI - Developing the marketing mix. PMID- 15117058 TI - Pseudoexfoliation syndrome and pseudoexfoliative glaucoma. AB - BACKGROUND: Pseudoexfoliative glaucoma (PXG) is a common secondary glaucoma that occurs when pigment and abnormal basement membrane material from anterior segment anatomical structures deposit in the trabecular meshwork. This anomaly arises from the predisposing condition, pseudoexfoliation syndrome (PXE). Complications are common, making this one of the most difficult glaucomas to manage. PURPOSE: The literature is reviewed in order to facilitate understanding of this condition. PMID- 15117060 TI - Selecting a business entity: more tax and non tax considerations. PMID- 15117061 TI - CMS to end 90-day grace period for discontinued HCPCS codes next year. PMID- 15117062 TI - [Audimont--a scientific research expedition to Mount Cho Oyu in the Himalayas]. AB - Even though hearing and vestibular disorders at high altitude have been reported since 1938, their reasons are still unknown. During the Audimont Research Expedition the risk for cochlear and vestibular sensory cells has been quantified by otoacoustic emissions and videonystagmography. Vestibular disorders could not be observed up to 7050 meters. However, the outer hair cells in the inner ear showed a reduction of emissions at increasing height. The pattern of the inner ear reply, depending on the altitude, complies with an increase of the perilymphatic pressure. As the perilymphatic space corresponds directly to the subarachnoid space, the limitation of hearing thus appears to be a direct consequence of raised intracranial pressure. PMID- 15117063 TI - [The effect of ginkgo biloba on healthy elderly subjects]. AB - BACKGROUND AND AIM: Over the past 25 years, numerous studies have confirmed the positive effect of the special ginkgo extract EGb 761 on the mental ability and emotional well-being of patients with cognitive disorders of vascular genesis, and Alzheimer-type dementia. The following study investigated the short-term effect of the special ginkgo extract EGb 761 on the subjective emotional well being of healthy elderly subjects. STUDY POPULATION AND METHOD: The study was designed as a randomized double-blind, monocenter study with parallel groups. It included 66 healthy subjects of both sexes aged between 50 and 65 with no age related cognitive impairments. For a period of 4 weeks, 34 subjects received a daily dose of 240 mg EGb 761, and 32 a placebo. Prior to starting medication and after 28 days of treatment, subjects completed the following scales and questionnaires to establish subjective emotional well-being: the Profile of Mood States (POMS), the Self Rating Depression Scale (SDS), three Visual Analog Scales to assess the quality of life (VAS-QoL), general health (VAS-GH) and mental health (VAS-MH), and a new instrument for assessing changes in general subjective well-being, the Subjective Intensity Score Mood (SIS Mood). Depending on the underlying distribution of the variables analyzed, parametric (t-tests) or nonparametric tests (U-tests) were performed to compare mean values and distributions both within and between the treatment groups. RESULTS: The final examination revealed a statistically significant difference between the two groups for the VAS mental health and quality of life, as also for SIS Mood at the telephone interview in week 2. A comparison of baseline with the final examination within the groups showed a statistically significant improvement in the EGb 761 group for the variables: depression, fatigue, anger and SDS. For none of the variables investigated was a worsening observed in the EGb 761 group. CONCLUSIONS: The results suggest a positive effect of EGb 761 on the subjective emotional well-being of healthy elderly persons. PMID- 15117064 TI - [Flupirtine in acute and chronic pain associated with muscle tenseness. Results of a postmarket surveillance study]. AB - BACKGROUND: Flupirtine with its specific mode of action as Selective Neuronal Potassium Channel Opener (SNEPCO) shows analgesic, muscle tone normalizing properties and iprotects from chronification. By means of an open multicentre drug monitoring study new data should be accumulated in terms of efficacy and patient's tolerance of flupirtine upon treatment of acute and chronic pain associated with muscle tenseness. RESULTS: Regarding acute and chronic pain, significant reduction of pain intensity, pain upon pressure and muscular tenseness could be proved. Simultaneously, decrease of sleep disorders due to pain and strong reduction of restrictions in daily life caused by pain improved distinctly patient's quality of life. Efficacy and tolerance has mainly been rated as good and very good respectively by physicians and patients, the number of undesired drug reactions being under 1%. CONCLUSION: Data of this study confirm the analgesic and muscle tone normalizing efficacy of flupirtine upon treatment of acute and chronic pain associated with muscle tenseness and show, that flupirtine has to be considered as a substance of first choice for the treatment of these disorders. PMID- 15117065 TI - [Treatment of chronic wounds with an alginate dressing containing calcium zinc and manganese]. AB - BACKGROUND: Treatment of chronic wounds comprises adequate debridement, reduction in the bacterial burden, and application of a dressing to absorb excess exudate. METHODS: In a prospectively documented, multicenter observational study involving 1285 chronic wounds of varying etiologies, carried out in 314 doctor's offices in Germany, the effect of the new alginate dressing, containing calcium zinc and manganese was investigated. RESULTS: After twelve weeks of treatment with the dressing, 95% of all wounds had improved appreciably, and 52.5% had healed completely. The drop out rate was 4.4% and side effects were observed in 2.6% of the cases. 73.8% of the patients reported markedly improved tolerability in comparison with previous treatment. 604 patients (47.0%) considered handling of the dressing to be "much better", and 411 (32.0%) to be "better" than previous treatment. The frequency of dressing changes decreased from 5.2 to 3.2 a week. PMID- 15117066 TI - [Antihypertensive first-line and add-on treatment with a fixed controlled release combination of metoprololsuccinate/hydrochlorothiazide. Prospective doctor's office observational study in 14,964 patients]. AB - AIM: To evaluate the efficacy and tolerability of a single daily dose of a fixed combination of 95 mg metoprololsuccinate (MS) and 12.5 mg hydrochlorothiazide (HCTZ) in the first-line treatment of non-pretreated hypertensives, or additional (add-on) to ongoing antihypertensive medication. METHOD: 14,964 patients aged 18 years or older treated by 2808 family doctors in Germany were included in a noncontrolled observational study. Most patients had at least one concurrent disease or concomitant medication of one kind or another. The primary target parameters for efficacy was the lowering of the systolic blood pressure (SBP) and diastolic blood pressure (DBP) after 8 weeks and for tolerability the number of patients reporting adverse events (AE). RESULTS: 65.4% of the patients received MS/HCTZ in the form of first-line treatment, the remainder as add-on therapy. The mean blood pressure decrease for the overall group by the end of the study was 24.5/-13.6 mmHg (baseline: 166.7/97.3 mmHg; p < 0.0001 for SBP and DBP). 92.2% of the patients experienced a decrease in SBP of > or = 10 mmHg. The mean heart rate decreased by 10.2 beats (baseline 81.4; p < 0.0001). The blood pressure decreased both in patients receiving MS/HCTZ alone and in those receiving it as an add-on to other antihypertensives. Only 1.4 of the patients reported AE. CONCLUSION: The MS/HCTZ controlled release combination was safe, efficacious and well-tolerated both as first-line and add-on therapy for essential hypertension. PMID- 15117067 TI - Purge-and-trap capillary gas chromatography with atomic emission detection for volatile halogenated organic compounds determination in waters and beverages. AB - A method for the simultaneous determination of 10 volatile halogenated organic compounds (VHOCs), including four trihalomethanes (THMs), in waters and beverages was developed. The analytes were stripped from the aqueous sample by a flow of helium, preconcentrated in a capillary trap and thermally desorbed using a purge and-trap (PT) system. This was followed by capillary gas chromatography with microwave-induced plasma atomic emission spectrometry (GC-AED). For element specific detection, three wavelengths were monitored, corresponding to chlorine (479 nm), bromine (478 nm) and iodine (193 nm). Each chromatographic run took 21 min, including the purge time. After careful choice of the experimental conditions, the performance of the system was evaluated. Calibration curves were obtained by plotting peak area versus concentration and the correlation coefficients for linear calibration were at least 0.9987. Detection limits, calculated for 5 ml sample volume, ranged from 0.05 microg l(-1) for chloroform to 0.5 microg l(-1) for tetrachloromethane. The method was successfully applied to the quantitative analysis of water samples of different origin and also of several beer and juice samples. The tap water samples analyzed contain variable concentrations of the four trihalomethanes, ranging from 1.0 to 66.5 microg l( 1), depending of the compound. Whereas chloroform, bromodichloromethane and bromoform were found in some of the juice samples, only chloroform was detected in the beer samples. The method is reliable and can be used for routine monitoring in water and beverages. PMID- 15117068 TI - Optimizing the determination of haloacetic acids in drinking waters. AB - Three methods are currently approved by the US Environmental Protection Agency for the compliance monitoring of haloacetic acids in drinking waters. Each derivatizes the acids to their corresponding esters using either acidic methanol or diazomethane. This study was undertaken to characterize the extent of methylation of these analytes by these methods, and to fully optimize methylation chemistries to improve analytical sensitivity, precision and accuracy. The approved methods were shown to have little to no esterification efficiencies for the brominated trihaloacetic acids (HAA3). Methylation with acidic methanol was determined to be more efficient and rugged than methylation with diazomethane. A new higher boiling solvent, tertiary-amyl methyl ether, is reported which has significantly improved methylation efficiencies for HAA3. Additional modifications to the method have been made that improve method ruggedness. The revised method, EPA Method 552.3, outperforms the currently approved methods, especially for HAA3. PMID- 15117069 TI - Determination of benzene, toluene, ethylbenzene and xylenes in soils by multiple headspace solid-phase microextraction. AB - Multiple headspace-solid phase microextraction (MHS-SPME) is a recently developed technique for the quantification of analytes in solid samples that avoids the matrix effect. This method implies several consecutive extractions from the same sample. In this way, the total area corresponding to complete extraction can be directly calculated as the sum of the areas of each individual extraction when the extraction is exhaustive, or through a mathematical equation when it is not exhaustive. In this paper, the quantitative determination of benzene, toluene, ethylbenzene and xylene isomers (BTEX) in a certified soil (RTC-CRM304, LGC Promochem) and in a contaminated soil by multiple HS-SPME coupled to a gas chromatography-flame ionisation detector (GC-FID) is presented. BTEX extraction was carried out using soil suspensions in water at 30 degrees C with a 75 microm carboxen-polydimethylsiloxane (CAR-PDMS) fibre and calibration was carried out using aqueous BTEX solutions at 30 degrees C for 30 min with the same fibre. BTEX concentration was calculated by interpolating the total peak area found for the soils in the calibration graphs obtained from aqueous solutions. The toluene, ethylbenzene, o-xylene and m,p-xylene concentrations obtained were statistically equal to the certified values. PMID- 15117070 TI - Reversed-phase retention thermodynamics of pure-water mobile phases at ambient and elevated temperature. AB - The use of pure water at superheated temperatures, between 100 and 200 degrees C, as a mobile phase for reversed-phase separations is explored. The thermodynamics of the retention process at low temperature (15-55 degrees C) are compared to the thermodynamics at elevated temperature (125-175 degrees C). Significant differences in the enthalpy of the retention process are observed between the two temperature ranges. This is possibly due to changes in the hydrogen-bond network of the pure-water mobile phase, which would change the solvation, and therefore retention, of non-polar solutes. The change in thermodynamic values between the two temperature regions invalidates extrapolation of retention as a function of temperature between the two temperature regions for the prediction of room temperature pure-water retention factors. The thermodynamic changes observed as the temperature is increased are similar to those seen when mobile phase composition is changed (by adding organic modifier) at constant temperature. PMID- 15117071 TI - Prediction of soil organic carbon partition coefficients by soil column liquid chromatography. AB - To avoid the limitation of the widely used prediction methods of soil organic carbon partition coefficients (KOC) from hydrophobic parameters, e.g., the n octanol/water partition coefficients (KOW) and the reversed phase high performance liquid chromatographic (RP-HPLC) retention factors, the soil column liquid chromatographic (SCLC) method was developed for KOC prediction. The real soils were used as the packing materials of RP-HPLC columns, and the correlations between the retention factors of organic compounds on soil columns (ksoil) and KOC measured by batch equilibrium method were studied. Good correlations were achieved between ksoil and KOC for three types of soils with different properties. All the square of the correlation coefficients (R2) of the linear regression between log ksoil and log KOC were higher than 0.89 with standard deviations of less than 0.21. In addition, the prediction of KOC from KOW and the RP-HPLC retention factors on cyanopropyl (CN) stationary phase (kCN) was comparatively evaluated for the three types of soils. The results show that the prediction of KOC from kCN and KOW is only applicable to some specific types of soils. The results obtained in the present study proved that the SCLC method is appropriate for the KOC prediction for different types of soils, however the applicability of using hydrophobic parameters to predict KOC largely depends on the properties of soil concerned. PMID- 15117072 TI - Direct high-performance liquid chromatographic method for enantioselective and diastereoselective determination of selected pyrethroic acids. AB - This study reports on the direct HPLC stereoisomer separation of selected pyrethroic acids employing commercial cinchona alkaloid derived chiral stationary phases (CSPs). cis/trans-Chrysanthemic acid (cis/trans-CA), cis/trans chrysanthemum dicarboxylic acid (cis/trans-CDCA), cis/trans-permethrinic acid (cis/trans-PA), and fenvaleric acid (FA) were resolved into the individual stereoisomers, i.e. enantiomers and diastereomers as well. To achieve satisfactory baseline separation an optimisation of the variables of the chromatographic method including chemical structure of the cinchona carbamate CSP, mobile phase composition, and flow rate was required. All four stereoisomers of PA were successfully separated in a single run (alphacis = 1.20, alphatrans = 1.26, critical Rs = 1.65) with an acetonitrile (ACN)-based polar-organic eluent. The complete baseline resolution of all CA stereoisomers succeeded in polar organic (alphacis = 1.20, alphatrans = 1.35, critical Rs = 3.03) as well as in acetonitrile-based reversed-phase media (alphacis = 1.24, alphatrans = 1.22, critical Rs = 2.73). The latter elution mode was also found to be suitable for the enantio- as well as diastereoselective resolution of CDCA (alphacis = 1.09, alphatrans = 1.50, critical Rs = 1.43), which is to the best of our knowledge the first reported enantiomer separation of this analyte. The enantiomers of FA could be baseline separated employing also reversed-phase mode (alpha = 1.16, Rs = 2.91). These separation methods may be applied for quality control processes in the production of stereoisomerically pure insecticides as well as stereoselective toxicokinetic studies, as CDCA, PA, and FA are suitable biomarkers for monitoring human pyrethroid burden. PMID- 15117073 TI - Application of preparative high-speed counter-current chromatography for the separation of flavonoids from the leaves of Byrsonima crassa Niedenzu (IK). AB - The methanolic extract of the leaves of the medicinal plant Byrsonima crassa (Malpighiaceae) contain flavonoids with antioxidant activity. They were separated in a preparative scale using high-speed counter-current chromatography. The optimum solvent system used was composed of a mixture of ethyl acetate-n-propanol water (140:8:80 (v/v/v)) and led to a successful separation between monoglucosilated flavonoids (quercetin-3-O-alpha-L-arabinoside, quercetin-3-O beta-D-galactoside) and the biflavonoid amentoflavone in only 3.5 h. The purities of quercetin-3-O-alpha-L-arabinoside (95 mg), quercetin-3-O-beta-D-galactoside (16 mg) and the biflavonoid amentoflavone (114 mg) were all isolated at purity over 95%. Identification was performed by 1H NMR, 13C NMR and UV analyses. PMID- 15117074 TI - Analysis of iso-alpha-acids and reduced iso-alpha-acids in beer by direct injection and liquid chromatography with ultraviolet absorbance detection or with mass spectrometry. AB - A liquid chromatography (LC) method is described for the simultaneous analysis of iso-alpha-acids and reduced iso-alpha-acids in beer. Volatile mobile phase additives were selected to enable hyphenation to mass spectrometric (MS) operated in the atmospheric pressure chemical ionization (APCI) mode. Contrary to other recent LC optimization procedures for the same compounds, an alkaline pH was selected hereby improving peak shape and selectivity. Both UV and MS detection are sensitive enough to analyze beers without sample pre-concentration. All major bitter acids are separated within 65 min with exception of cis dihydroisoadhumulone, which co-elutes with trans-isocohumulone. Due to the selectivity of the MS, these compounds could be differentiated according to their m/z value. The performance in terms of quantification of bitter acids by LC-UV and LC-MS are compared for standard solutions and a selection of 14 beers. PMID- 15117075 TI - Simultaneous separation and quantitation of amino acids and polyamines of forest tree tissues and cell cultures within a single high-performance liquid chromatography run using dansyl derivatization. AB - The objective of the present study was to develop a rapid HPLC method for simultaneous separation and quantitation of dansylated amino acids and common polyamines in the same matrix for analyzing forest tree tissues and cell cultures. The major modifications incorporated into this method as compared to previously published HPLC methods for separation of only dansyl amino acids include: use of a 10 cm column to reduce the total run time by approximately 15 min; modification of the dansyl derivatization process and gradient profile to elute amino acids and common polyamines within the same run; addition of steps for column cleaning within each run; shorter re-equilibration time; and finally, column cleaning and physically reversing the column at the end of a loop of samples. These changes improved peak resolution and increased column longevity by several-fold. Over 1000 foliar samples from mature forest trees could be analyzed with the same column as compared to only 200-250 samples before the incorporation of these changes. This method eluted 22 amino acids within 40 min plus all three common polyamines between 44 and 47 min. The total run time is 53.6 min for amino acids only and 55.6 min for both amino acids and polyamines. PMID- 15117076 TI - Sensitive determination of aliphatic amines in water by high-performance liquid chromatography with chemiluminescence detection. AB - A sensitive method has been developed for liquid chromatographic determination of short aliphatic amines in water samples. Analytes are preconcentrated and dansylated on solid sorbents (C18 solid-phase extraction cartridges). The dansyl derivatives are chromatographed and post-column mixed with peroxyoxalate (TCPO) and H2O2 in order to perform chemiluminescence detection. Optimal results have been obtained using a sample volume of 5 ml. The method has been applied to the quantification or screening of several aliphatic amines: methylamine, ethylamine, butylamine, diethylamine, pentylamine and hexylamine. The screening procedure has been developed including also polyamines (putrescine, cadaverine, spermidine and spermine). The results obtained by using chemiluminescence (CL) detection have been compared with other detection systems (fluorescence and UV). The sensitivity can increase from 3 to 75 times respect UV detection and from 2 to 10 times respect fluorescence detection depending on the amine. The detection limits achieved were between 0.15 and 0.9 microg/l. PMID- 15117077 TI - Purification of recombinant green fluorescent protein by three-phase partitioning. AB - The technique of three-phase partitioning (TPP) was used to purify the green fluorescent protein (GFP) in a single step. TPP uses a combination of ammonium sulphate and tert-butanol to precipitate proteins from their crude extracts. In the first round of TPP with 20% ammonium sulphate saturation at the ratio of crude to tert-butanol 1:1 (v/v), most of the GFP remains in the lower aqueous phase. When subjected to a second round of TPP with 60% ammonium sulphate saturation at the ratio of crude to tert-butanol 1:2 (v/v) gives 78% recovery of GFP with a 20-fold purification. The sodium dodecyl sulphate-polyacrylamide gel electrophoretic (SDS-PAGE) analysis of purified preparation shows single band. The fluorescence excitation and emission spectra agreed with values reported in literature. PMID- 15117078 TI - Fluorimetric determination of pantothenic acid in foods by liquid chromatography with post-column derivatization. AB - A method to determine the content of free pantothenic acid in various foods by reverse phase liquid chromatography-fluorimetry is reported. It includes a purification of the samples by successive passages through anion and cation exchange cartridges and a post-column derivatization of pantothenic acid as the fluorescent 1-alkylthio-2-alkylisoindole (reaction of beta-alanin, formed by hot alkaline hydrolysis of pantothenic acid, with orthophthaldialdehyde in the presence of 3-mercaptopropionic acid). An enzymatic hydrolysis prior to the purification step (pepsin at 50 degrees C for 3 h, then pantetheinase and alkaline phosphatase at 20 degrees C for 18 h) made it possible to release the bound pantothenic acid and thus to obtain the total Vitamin B5 content of these foodstuffs. The method proposed for the determination of free and bound pantothenic acid gives a good recovery rate (96-101%) and a satisfactory repeatability (R.S.D.r less than 8%). Owing to its low detection limit (0.65 microg g(-1)) and the good resolution of the pantothenic acid peak, it could most probably be applied to the determination of this vitamin in any foodstuff. PMID- 15117080 TI - Simultaneous determination of alkylphenol ethoxylates and their biotransformation products by liquid chromatography/electrospray ionisation tandem mass spectrometry. AB - Reversed-phase LC-MS/MS is used to determine major estrogenic alkylphenol ethoxylates (APEOs) and their biotransformation products. It allows the simultaneous analysis of eight APEOs, alkylphenoxy carboxylates (APECs) and alkylphenols (APs) in sewage treatment plant (STP) effluents in the same extract after solid-phase enrichment on polymeric Oasis HLB. As precursor ions, [APEO + NH4]+, [APEC - H]- and [AP - H]- were monitored. Instrumental limits of detection (LOD) were 2-600 pg, corresponding to sample concentrations of 0.04-12 ng l(-1), without correction for overall method recoveries. Matrix-induced signal suppression during electrospray ionisation (ESI) and extraction as well as overall method recoveries were assessed and the suitability of deuterated surrogates as internal standards was evaluated. PMID- 15117079 TI - Characterisation of botulinum toxins type C, D, E, and F by matrix-assisted laser desorption ionisation and electrospray mass spectrometry. AB - In a follow-up of the earlier characterisation of botulinum toxins type A and B (BTxA and BTxB) by mass spectrometry (MS), types C, D, E, and F (BTxC, BTxD, BTxE, BTxF) were now investigated. Botulinum toxins are extremely neurotoxic bacterial toxins, likely to be used as biological warfare agent. Biologically active BTxC, BTxD, BTxE, and BTxF are comprised of a protein complex of the respective neurotoxins with non-toxic non-haemagglutinin (NTNH) and, sometimes, specific haemagglutinins (HA). These protein complexes were observed in mass spectrometric identification. The BTxC complex, from Clostridium botulinum strain 003-9, consisted of a 'type C1 and D mosaic' toxin similar to that of type C strain 6813, a non-toxic non-hemagglutinating and a 33 kDa hemagglutinating (HA 33) component similar to those of strain C-Stockholm, and an exoenzyme C3 of which the sequence was in full agreement with the known genetic sequence of strain 003-9. The BTxD complex, from C. botulinum strain CB-16, consisted of a neurotoxin with the observed sequence identical with that of type D strain BVD/-3 and of an NTNH with the observed sequence identical with that of type C strain C Yoichi. Remarkably, the observed protein sequence of CB-16 NTNH differed by one amino acid from the known gene sequence: L859 instead of F859. The BTxE complex, from a C. botulinum isolated from herring sprats, consisted of the neurotoxin with an observed sequence identical with that from strain NCTC 11219 and an NTNH similar to that from type E strain Mashike (1 amino acid difference with observed sequence). BTxF, from C. botulinum strain Langeland (NCTC 10281), consisted of the neurotoxin and an NTNH; observed sequences from both proteins were in agreement with the gene sequence known from strain Langeland. As with BTxA and BTxB, matrix-assisted laser desorption/ionisation (MALDI) MS provided provisional identification from trypsin digest peptide maps and liquid chromatography electrospray (tandem) mass spectrometry (LC-ES MS) afforded unequivocal identification from amino acid sequence information of digest peptides obtained in trypsin digestion. PMID- 15117081 TI - Trace level determination of acrylamide in cereal-based foods by gas chromatography-mass spectrometry. AB - A quantitative method has been developed for the determination of trace levels (<50 microg/kg) of acrylamide in cereal-based foods. The method is based on extraction of acrylamide with water, acidification and purification with Carrez I and II solutions, followed by bromination of the acrylamide double bond. The reaction product (2,3-dibromopropionamide) is extracted with ethyl acetate/hexane (4:1, v/v), dried over sodium sulfate, and cleaned up through a Florisil column. The derivative is then converted to 2-bromopropenamide by dehydrobromination with triethylamine and analyzed by gas chromatography coupled to mass spectrometry (GC MS), employing (13C3)acrylamide as internal standard. In-house validation data for commercial and experimental cereal products showed good precision of the method, with repeatability and intermediate reproducibility relative standard deviations below 10%. The limit of detection and limit of quantitation are estimated at 2 and 5 microg/kg, respectively, and recoveries of acrylamide from samples spiked at levels of 5-500 microg/kg ranged between 93 and 104% after correction of analyte loss by the internal standard. Finally, a comparative test organized with two independent laboratories provided additional confidence in the good performance of the method, particularly at very low concentration levels. PMID- 15117082 TI - Determination of ionization constants of N-imidazole derivatives, aromatase inhibitors, using capillary electrophoresis and influence of substituents on pKa shifts. AB - Capillary electrophoresis (CE) was used as a method to determine the acidity constants of eight aromatase inhibitors. This method was validated by comparison of results obtained with a traditional method, UV spectroscopy, and additionally with computational calculations. We confirmed here, with our series of compounds, that capillary electrophoresis is an attractive method for pKa measurements which is based on migration time or mobilities of the ionic species over a range of pH values. The precision of pKa measurements of N-imidazole derivatives is useful to observe pKa shifts induced by chemical modifications introduced on adjacent aromatic rings such as heterocycle (benzoxa- or benzothiazolinone) or substituted benzyle. The knowledge of these pKa values is a great interest to predict migration of solutes and qualitative interactions with ionized cyclodextrines as chiral selectors in further enantioseparative CE studies. PMID- 15117083 TI - Development of a novel analytical method for determination of chondroitin sulfate using an in-capillary enzyme reaction. AB - A novel analytical method for determination of total amount of chondroitin sulfate (CS) based on its conversion to desulfated chondro-disaccharide via an enzyme-catalyzed reaction, was developed. Using the in-capillary enzyme reaction, the method was also applied to the successful construction of an on-line analytical system. Within this system, electrophoretic migration was used to mix zones containing the enzyme mixture (chondroitinase ABC, chondro-4-sulfatase, chondro-6-sulfatase and 2-o-sulfatase) and the substrate (CS). The reaction was then allowed to proceed in the presence of a weak electric field and, finally, the product (desulfated chondro-disaccharide) of enzyme reaction migrated to the detector under the influence of an applied electric field. A polyvinyl alcohol coated capillary was used to reduce protein adsorption. Desulfated chondro disaccharide was successfully migrated toward the anode in 10 mM Tris-acetate buffer (pH 7.0) under reversed polarity and detected at 232 nm. The established method was validated and demonstrated to be applicable in the determination of total amount of CS in a commercial ophthalmic solution. No interference from the formulation excipients was observed. Good linearity was obtained, with correlation coefficients above 0.999. Recoveries and precisions ranged from 100.0 to 100.5%, and from 0.2 to 0.6% of the relative standard deviation, respectively. Good agreement was obtained between the established method and traditional photometric method based on carbazole reaction. In this study, application of the method to disaccharide compositional analysis was also performed. PMID- 15117084 TI - Simultaneous determination of iodide and iodate in seawater by transient isotachophoresis-capillary zone electrophoresis with artificial seawater as the background electrolyte. AB - We developed capillary zone electrophoresis with transient isotachophoresis (ITP) as an on-line concentration procedure for simultaneous determination of iodide and iodate in seawater. The effective mobility of iodide was decreased by addition of 20 mM cetyltrimethylammonium chloride to an artificial seawater background electrolyte so that transient ITP functioned for both iodide and iodate. Limits of detection for iodide and iodate were 4.0 and 5.0 microg/l (as iodine) at a signal-to-noise ratio of 3. Values of the relative standard deviation of peak area, peak height, and migration times for iodide and iodate were 2.9, 1.3, 1.0 and 2.3, 2.1, 1.0%, respectively. The proposed method was applied to simultaneous determination of iodide and iodate in seawater collected at a pond at our university. PMID- 15117085 TI - Weight adjustment in calculating surface ligand coverage or density for chromatographic bonded phases. AB - A method based on successive approximation is proposed for the calculation of surface coverage or ligand density of chemically bonded stationary phases from elemental analysis data. The approach could be used to calculate surface coverage in many stationary phase preparations, including end-capping and stepwise couplings. Key in this procedure is the use of successive approximation to simplify the adjustment of weight changes in stationary phase preparation. The method does not involve complicated mathematic equations. PMID- 15117086 TI - Improved coupled-column liquid chromatographic method for the determination of glyphosate and aminomethylphosphonic acid residues in environmental waters. AB - An existing method for the determination of glyphosate and its main metabolite aminomethylphosphonic acid (AMPA) in water has been improved. It is based on precolumn derivatization with the fluorescent reagent 9 fluorenylmethylcloroformate (FMOC) followed by large-volume injection in a coupled-column LC system using fluorescence detection (LC-LC-FD). The derivatization step was slightly modified by changing parameters such as volume and/or concentration of sample and reagents to decrease the limits of quantification (LOQ) of glyphosate and AMPA to 0.1 microg/l. Additionally, the use of Amberlite IRA-900 for preconcentration of glyphosate, prior to the derivatization step, was investigated; the LOQ of glyphosate was lowered to 0.02 microg/l. Drinking, surface and ground water spiked with glyphosate and AMPA at 0.1-10 microg/l concentrations were analysed by the improved LC-LC-FD method. Recoveries were 87-106% with relative standard deviations lower than 8%. Drinking and ground water spiked with glyphosate at 0.02 and 0.1 microg/l were analysed after preconcentration on the anion-exchange resin with satisfactory recoveries (94-105%) and precision (better than 8%). PMID- 15117087 TI - Vardenafil--PDE5 inhibitor number 3. PMID- 15117088 TI - Sustained efficacy and tolerability with vardenafil over 2 years of treatment in men with erectile dysfunction. AB - This randomised, double-blind study assessed the long-term efficacy and tolerability of vardenafil 10 and 20 mg in men with erectile dysfunction (ED). A total of 566 men who completed an initial 12-month treatment period entered a 12 month extension. In these men, both doses of vardenafil produced improvement in scores for the 'erectile function' Domain of the International Index of Erectile Function, evident from week 4 and maintained through 2 years. Sexual Encounter Profile diary responses indicated that following treatment, penetration was achieved on 92-94% of attempts and erections that lasted long enough for successful intercourse were achieved on 87-89% of attempts. In response to the General Assessment Question, 90-92% of patients reported improved erections with vardenafil. Most treatment-emergent events were mild and transient with no cardiovascular safety concerns. These results support the long-term efficacy, reliability and tolerability of vardenafil 10 and 20 mg in men with ED. PMID- 15117089 TI - Urinary beta-2 microglobulin in renal dysfunction associated with hypothyroidism. AB - Hypothyroidism is associated with a number of functional renal disorders primarily affecting water and salt handling. In this study, we aim to investigate functional renal disorders in overt and subclinical hypothyroidism. We utilised urinary beta-2 microglobulin (beta2M) as a sensitive marker of tubular dysfunction. Urinary beta2M excretion and tests for renal functions were repeated before and after T4 replacement therapy. Forty-four patients (28 with overt disease, 16 with subclinical disease) and 31 healthy control subjects were involved in the study. There was a significant increase in urinary beta2M in both overt and subclinical hypothyroidism (p < 0.05 in both). TSH levels were correlated with beta2M excretion. Effects of hypothyroidism on renal functions were readily reversible by 3 weeks of thyroid hormone replacement therapy. Urinary beta2M was a sensitive marker of renal tubular dysfunction associated with overt and subclinical hypothyroidism. PMID- 15117090 TI - Analgesic efficacy of a single dose of lumiracoxib compared with rofecoxib, celecoxib and placebo in the treatment of post-operative dental pain. AB - This randomised, double-blind, placebo-controlled, parallel-group study compared the efficacy and tolerability of lumiracoxib (a novel COX-2 selective inhibitor) with rofecoxib, celecoxib and placebo in patients with moderate-to-severe post operative dental pain. Following third molar extraction, patients received single oral doses of lumiracoxib 400 mg, rofecoxib 50 mg, celecoxib 200 mg or placebo (n = 355). Additional patients from a similar study, assigned to lumiracoxib, rofecoxib or placebo (n = 155), were included for analysis of the primary variable, Summed Pain Intensity Difference over the first 8 h post dose (SPID-8). For SPID-8, lumiracoxib was superior to rofecoxib (p < 0.05), celecoxib (p < 0.001) and placebo (p < 0.001). Lumiracoxib demonstrated the fastest onset of analgesia and the longest time to rescue medication use. Patient global evaluation of lumiracoxib was comparable to rofecoxib and superior to celecoxib and placebo. All treatments were well tolerated. Lumiracoxib 400 mg provides rapid, effective and sustained relief of post-operative dental pain, comparable or superior to rofecoxib. PMID- 15117091 TI - Analgesic efficacy of single oral doses of lumiracoxib and ibuprofen in patients with postoperative dental pain. AB - This randomised, double-blind study compared single dose lumiracoxib (a cyclooxygenase-2 selective inhibitor) 100 and 400 mg, ibuprofen 400 mg and placebo in patients with postoperative dental pain over 12 h. The primary efficacy variable was pain intensity difference. Lumiracoxib 400 mg and ibuprofen were superior to placebo from 1 to 12 h post dose while lumiracoxib 100 mg was superior from 1.5 to 9 h. Lumiracoxib 400 mg demonstrated the fastest median time to onset of analgesia (37.4 min) followed by ibuprofen (41.5), and lumiracoxib 100 mg (52.4; all p < or = 0.001 vs. placebo). Median time to rescue medication (h) was longer for lumiracoxib 400 mg (> or = 12), lumiracoxib 100 mg (approximately 7) and ibuprofen (approximately 8) than placebo (approximately 2; all p < or = 0.001 vs. placebo). Patients rated lumiracoxib 400 mg superior to the other active treatments (p < 0.05); lumiracoxib 100 mg was comparable with ibuprofen and superior to placebo (p < 0.001). Lumiracoxib provided rapid, effective and well-tolerated analgesia. PMID- 15117092 TI - What role do community pharmacists currently play in the management of headache? A hospital-based perspective. AB - Of one hundred consecutive patients with headache referred by general practitioners to general neurology outpatient clinics, only 15 had at any time consulted a community pharmacist for advice about headache or its treatment; none spontaneously volunteered this information. Community pharmacists may represent an underused resource in the management of headaches in primary care, but would require specific training to fulfil such a role. PMID- 15117093 TI - Combined use of autologous transfusion techniques to avoid allogeneic transfusion in spinal fusion surgery with instrumentation. AB - This study conducted a retrospective review of the medical records of 321 patients to delineate the efficacy of the combined use of autologous transfusion (AT) techniques. Transfusion profiles between an AT and homologous transfusion (HT) group were compared. A much lower proportion of patients were exposed to allogeneic blood in the AT group (13%) than in the HT group (98%, p<0.001). In the AT group, a significantly smaller proportion of patients were exposed to allogeneic blood in patients transfused with three or four AT techniques (8%) than those with one or two techniques (29%, p<0.05). A febrile reaction (11% of patients) after a reinfusion of post-operatively shed blood was the only side effect associated with an AT. In conclusion, an AT is effective for preventing the exposure of allogeneic blood in spinal fusion surgery. The combined use of multiple AT techniques may further improve its efficacy. PMID- 15117094 TI - Community pharmacists' knowledge and dispensing recommendations for treatment of acute diarrhoea in Trinidad, West Indies. AB - Using a hypothetical case presentation of a patient with acute diarrhoea, community pharmacists in Trinidad were asked about their knowledge and dispensing recommendations to manage acute diarrhoea. Oral rehydration salts (ORS) were recommended by 86% (79), but more pharmacists would recommend ORS as the first choice therapy alone, for children (70%) than adults (33%) (p < 0.01). Antimotility agents as a first choice therapy alone or with ORS would be given to more adults (60%) than children (10%) (p < 0.01), and more adults (59%) than children (33%) would receive cotrimoxazole. Pharmacists (93%) would counsel on preparation, storage and treatment schedule for ORS, but not on discontinuing (32%) or continuing ORS (4%). Despite 51 pharmacists knowing the WHO guidelines to treat acute diarrhoea, only 23 dispensed in accordance. Educational re enforcement to manage acute diarrhoea and dispensing practices of medications are necessary for pharmacists who are the first patient contact in Trinidad. PMID- 15117095 TI - What is the impact of PRIME on real-life diabetic nephropathy? AB - Type 2 diabetes is increasing globally and is a major cause of conditions such as cardiovascular disease, retinopathy and nephropathy. The Diabetes Control and Complications Trial and the UK Prospective Diabetes Study demonstrated that the progression of renal disease could be slowed by tight glycaemic control and treating any associated hypertension with angiotensin-converting enzyme inhibition. Recent clinical trials have supported the use of angiotensin II receptor antagonists in the treatment of diabetic nephropathy, resulting in the approval of new therapeutic indications in the United States and Europe. The objective of this review is to demonstrate how results from the Program for Irbesartan Mortality and morbidity Evaluation studies apply to clinical practice, and to show how the benefits of irbesartan therapy can be realised at any stage of renal disease in patients with diabetes. PMID- 15117096 TI - Current perspectives on the treatment of venous thromboembolism: need for effective, safe and convenient new antithrombotic drugs. AB - Treatment of venous thromboembolism (VTE) has evolved significantly over the last decade. Low-molecular-weight heparins have largely replaced unfractionated heparin in the treatment of deep-vein thrombosis (DVT) but the majority of patients with pulmonary embolism (PE) continue to be treated with unfractionated heparin. Fondaparinux is the first synthetic selective inhibitor of factor Xa. It has recently been proved to be more effective than, and as safe as, a low molecular-weight heparin for the prevention of VTE after major orthopaedic surgery. The two large randomised MATISSE trials demonstrated that fondaparinux was at least as effective and as safe as previous reference heparin therapies in the treatment of VTE. Fondaparinux should further simplify the treatment of this frequent disease since a single once-daily fixed dosage regimen may effectively and safely treat both DVT and PE, an important point especially considering the frequent though clinically silent concomitance of these two thrombotic events. PMID- 15117097 TI - Bupropion as an aid to smoking cessation: a review of real-life effectiveness. AB - Clinical trials rigorously demonstrate the efficacy of new products and justify their marketing. However, it is only after use in real-life settings that the clinical value (effectiveness) of a new treatment is fully known. The purpose of this review was to summarise the effectiveness data for bupropion SR as an aid to smoking cessation. Available reports of effectiveness data for bupropion SR were obtained from the literature, presentations at smoking cessation meetings and from the manufacturer. Twelve sources of effectiveness data were found and included clinical practice trials, observational studies/surveys, motivational support programme results and employer-based cessation programme results. The 6 month point prevalence smoking cessation rates ranged from 25 to 49%. There is a growing body of evidence supporting the effectiveness of bupropion SR as an aid to smoking cessation. Real-life quit rates for bupropion SR are similar to those seen in the original clinical trial programme. PMID- 15117098 TI - Fatal meningitis due to Listeria monocytogenes in elderly patients with underlying malignancy. AB - Adult patients with malignancies are considered to be at a high risk for Listeria monocytogenes meningitis. The Microbiology Laboratory's database of the University Hospital of Ioannina, Greece, was searched for cases of L. monocytogenes during the period from January 1990 to December 2002. Listerial meningitis occurred in three patients: one with brain tumour, one with chronic lymphocytic leukaemia, and one with non-Hodgkin's lymphoma. All the patients were older than 70 and they were actively receiving therapy for their malignancy. L. monocytogenes type 4b was isolated from blood and cerebrospinal fluid. All were treated with ampicillin and gentamicin, but they died shortly after the initiation of the treatment. Experience with the three present cases indicated the high mortality rate due to listerial meningitis in this immunosuppressed population. So, listeriosis should be suspected in patients with meningitis and underlying malignancy. Since meningitis due to L. monocytogenes is not distinguishable clinically from other types of bacterial meningitis, it is recommended to cover Listeria in the initial empirical therapy of bacterial meningitis in immunosuppressed patients. PMID- 15117099 TI - Eicosapentaenoic acid-rich essential fatty acid supplementation in chronic fatigue syndrome associated with symptom remission and structural brain changes. AB - Lateral ventricular enlargement has been reported in chronic fatigue syndrome, while cerebral neurospectroscopy has recently indicated that essential fatty acid treatment may be of value in this condition. An essential fatty acid supplement rich in eicosapentaenoic acid (EPA) was therefore given daily to a female patient with a 6-year history of unremitting symptoms of chronic fatigue syndrome. Cerebral magnetic resonance scanning was carried out at baseline and 16 weeks later. The EPA-rich essential fatty acid supplementation led to a marked clinical improvement in her symptoms of chronic fatigue syndrome, starting within 6-8 weeks. Accurate quantification of the lateral ventricular volumes in the baseline and 16-week follow-up registered images of high-resolution magnetic resonance imaging structural scans showed that the treatment was accompanied by a marked reduction in the lateral ventricular volume during this period, from 28,940 23,660 mm3. PMID- 15117100 TI - An analysis of an open access general practitioner bone densitometry service. AB - General practitioners (GPs) are increasingly involved in the selective opportunistic case finding of osteoporosis and treatment of this condition. An open access bone densitometry service has existed for GPs in the Cardiff area since 1993. The purpose of this study was to assess the effectiveness of this service, particularly with respect to the appropriateness of bone densitometry requests and the impact of the service on hospital outpatient referrals. Over a period of 12 months, 560 patients were referred to the open access service by 154 GPs. A total of 229 (41%) patients were given a clinical diagnosis of osteoporosis. Four hundred and seventy-three (84%) referrals followed the clinical referral criteria. In 85% of cases, GPs claimed that they would have referred patients to the hospital outpatient department, if there were no open access bone densitometry service. The majority of the referrals were appropriate, and the service appears to have reduced GP referrals to the specialist clinic. PMID- 15117101 TI - Mucin gene expression in cardiac myxoma. AB - Myxoma is the most common benign neoplasm of the heart. This work is the first to present an unusual left atrium and mitral valve cardiac myxoma which cannot be completely resected. This cardiac myxoma was also associated with abundant mucopolysaccharidic matrix, including mucin. Mucin gene expression is cell- and tissue-specific, with variations during cell differentiation and inflammation, and is altered during carcinogenesis. The expression of mucin genes in cardiac myxoma has never been elucidated previously. Detailed immunohistochemical analysis of MUC1, MUC2 and MUC5AC has been performed in this left atrium and mitral valve myxoma. Notably, the expressions of mucins in cardiac myxoma must be further evaluated. PMID- 15117102 TI - Pseudoaneurysm formation after flexible ureterorenoscopy and electrohydraulic lithotripsy. AB - Although electrohydraulic lithotripsy (EHL) is regularly used to fragment urinary calculi, it has the potential to cause significant renal tract damage. We report a complication of EHL use during flexible ureterorenoscopy, where recovery was complicated by delayed, prolonged haematuria secondary to the formation of a pseudoaneurysm in an interlobar renal artery. Embolisation was achieved by the insertion of a coil. PMID- 15117103 TI - Steroid responsiveness in a case of Riedel's thyroiditis and retroperitoneal fibrosis. AB - Riedel's thyroiditis is a rare chronic inflammatory disease of the thyroid characterised by an invasive fibrotic process. We present a lady with newly diagnosed hypothyroidism, rapidly enlarging hard, fixed goitre, strongly positive thyroid antibodies and raised erythrocyte sedimentation rate (ESR). A tru-cut biopsy confirmed Riedel's struma. Regression of the goitre and reduction of antibody titres occurred after starting prednisolone, which was stopped after 10 months. Six months later, she presented with renal failure due to retroperitoneal fibrosis that was successfully treated by reinstitution of steroids and ureteric stenting. Very high titres of thyroid antibodies and hypothyroidism predating development of goitre suggest coexistence of Hashimoto's thyroiditis and Riedel's thyroiditis. Tru-cut biopsy obviated the need for open thyroidectomy. A predominantly inflammatory as opposed to fibrotic thyroid histology may predict good response to steroids. Relapse following steroid withdrawal may not only be in the thyroid but also at other sites. PMID- 15117104 TI - Meningioma presenting with only urinary symptoms which is diagnosed by magnetic resonance imaging followed by urodynamic study. AB - Neurological tumour processes when involving bladder organs or innervations may give rise to urological symptoms. Depending on the organ or nerve tract affected, and emptying capacity of the bladder, the clinical signs manifested may be extremely varied, simulating different pathological entities that may lead to unnecessary treatment. PMID- 15117105 TI - Avulsion of the gallbladder following trivial injury. AB - Traumatic cholecystectomy is a rare condition that has always been described in the context of major trauma and associated liver or biliary injuries. We present a case of isolated traumatic cholecystectomy following a trivial injury which resulted in both a delayed presentation and a difficult diagnosis. PMID- 15117106 TI - Patients with migraine prefer zolmitriptan orally disintegrating tablet to sumatriptan conventional oral tablet. PMID- 15117107 TI - Epidemiology of postmenopausal hypertension. AB - Hypertension is more common in younger men than women but this trend is inverted at approximately 60 years of age--thereafter hypertension is more common in women. Menopause's contribution to this phenomenon is complex. Oestrogen deficiency after menopause precipitates a number of factors and these have established the 'menopausal metabolic syndrome' as a concept in postmenopausal women. However, studies have indicated that changes in the prevalence of hypertension, and overall cardiovascular risk profiles in postmenopausal women, might be due to ageing and not oestrogen deficiency. Undoubtedly, there is a strong multicolinearity between the two phenomena. Furthermore, hormone replacement therapy (HRT) may reduce age-induced blood pressure increases, thus decreasing cardiovascular risks. However, recent results have questioned HRT's role in cardiovascular disease (CVD) prevention in postmenopausal women and trials have unequivocally shown that CVD risk in postmenopausal women with hypertension can be effectively reduced by common antihypertensive drugs. PMID- 15117108 TI - Basic research into the mechanisms responsible for postmenopausal hypertension. AB - Women typically experience increased cardiovascular disease (CVD) following menopause. Major risk factors for CVD include hypertension and, after menopause, blood pressure increases in women. The mechanism(s) responsible for this increase are not determined. Changes in oestrogen/androgen ratios, possible activation of the renin-angiotensin system, and increases in endothelin and oxidative stress, are hallmarks in postmenopausal women. Obesity, type 2 diabetes and activation of the sympathetic nervous system may also play important roles. However, progress in clarifying the mechanisms responsible for postmenopausal hypertension has been hampered by lack of a suitable animal model. We have recently characterised the ageing female spontaneously hypertensive rat (SHR) as a model of postmenopausal hypertension, since this strain exhibits many of the humoral characteristics of postmenopausal women. This review discusses some of the mechanisms that could play a role in postmenopausal hypertension, as well as the characteristics of the ageing female SHR as a model. PMID- 15117109 TI - The metabolic syndrome in women: implications for therapy. AB - It is becoming increasingly clear that hypertension and metabolic risk factors in women are inter-related and often share underlying causes. Menopause acts explicitly as a risk factor by reducing the direct beneficial effect of ovarian hormones upon cardiovascular functions and indirectly by negatively influencing other risk factors for coronary artery disease--i.e. hyperinsulinaemia, blood cholesterol, blood pressure, coagulation etc. Adverse changes in one factor may induce adverse changes in a variety of other risk factors and it is important to consider co-ordinated changes when evaluating these patients rather than attempt to isolate independent factors. Similarly with treatment, the prevalence of metabolic syndrome in postmenopausal hypertensive women has important implications and some antihypertensive drugs may worsen the already altered metabolic profile of these patients while others may be beneficial. Centrally acting sympatholytic agents, e.g. moxonidine, are therefore important to consider in hypertensive postmenopausal women who experience other symptoms of metabolic syndrome. PMID- 15117110 TI - Treatment of postmenopausal hypertension with moxonidine, a selective imidazoline receptor agonist. AB - This study compared the effects of two sympatholytic agents--one central (moxonidine) and one peripheral (atenolol)--on blood pressure and other metabolic syndrome factors in postmenopausal hypertensive women who were not taking hormone replacement therapy. Atenolol and moxonidine led to a statistically significant reduction in diastolic blood pressure of 9.5 mmHg and 5.5 mmHg, respectively. A clear rebound effect was observed in the atenolol patients whereas the moxonidine group exhibited a slightly further decrease in blood pressure. Moxonidine also caused a profound decrease in both mean plasma-glucose area under the curve (AUC) during oral glucose tolerance test (-0.96 mmol/L x H, NS) and mean plasma-insulin AUC (-6.15 mU/L x H). Therefore, moxonidine displayed a slightly less potent antihypertensive effect than atenolol in hypertensive postmenopausal women, but it demonstrated a better metabolic effect. To conclude, moxonidine could benefit hypertensive postmenopausal women who display other signs of metabolic syndrome. PMID- 15117111 TI - The epidemiologist, genetics and system biology. PMID- 15117112 TI - Confounding and effect-measure modification in the evaluation of immunogenic agents. PMID- 15117114 TI - What do negative associations between potential risk factors and illness in analytical epidemiological studies of infectious disease really mean? AB - In epidemiological studies of infectious diseases it is not unusual to find that some potential risk factors are negatively associated with risk of illness. The mechanisms generating these associations are unclear in many cases, though one explanation is immunity due to prior exposure. We derive mathematical models for the proportion of a population who are infected with a disease and the proportion who are susceptible in any year of life when individuals are at risk of exposure through more than one route. It is shown that risk of illness declines with increasing age and that this risk declines most rapidly in those groups at increased exposure. In high exposure groups, the relative risk of illness, compared to a group with lower exposure, also declines with age, eventually becoming less than one. The threshold age at which the relative risk is less than 1, i.e., factor B becomes protective decreases with higher exposure rates. Epidemiological studies may substantially underestimate the importance of risk factors where exposure is consistent over many years. PMID- 15117113 TI - Genetic epidemiological approaches in the study of risk factors for cardiovascular disease. AB - The importance of genetic factors for the susceptibility to disease has been widely recognized in the last years. Genes have been identified for monogenic diseases and the challenge lying ahead is the identification of genetic components of importance and the environments in which they are expressed for complex diseases, that is, multiple genetic factors act and interact with each other or environmental factors to add to the complexity. This paper gives a brief overview of some genetic epidemiological approaches, concepts and recent methodological developments related to the study of risk factors for cardiovascular disease in twin and family studies. PMID- 15117115 TI - Increased risk for cardiovascular mortality in psoriasis inpatients but not in outpatients. AB - We have conducted a historical cohort study to assess cardiovascular mortality among psoriasis patients. Using the Swedish Inpatient Registry, we selected 8991 patients hospitalized for psoriasis at dermatological wards. To represent an outpatient cohort, 19,757 members of the Swedish Psoriasis Association were selected. Mortality from cardiovascular diseases was compared with the general population. We found no increased cardiovascular mortality among outpatients with psoriasis (standardized mortality ratio, SMR 0.94; 95% confidence interval, CI: 0.89-0.99). The overall risk among inpatients admitted at least once was increased by 50% (SMR 1.52; 95% CI: 1.44-1.60). The excess risk increased with increasing number of hospital admissions (p for trend <0.001). Cardiovascular mortality was higher among those admitted at younger ages (p for trend <0.001; SMR 2.62, 95% CI: 1.91-3.49, for patients aged 20 to 39 years at first admission). Young age at first admission appeared to further increase the risk among those who were repeatedly admitted. We conclude that a diagnosis of psoriasis per se does not appear to increase the risk for cardiovascular mortality. Severe psoriasis, however, here measured as repeated admissions, and early age at first admission, is associated with increased risk for cardiovascular death. PMID- 15117116 TI - Acute myocardial infarction population incidence and in-hospital management factors associated to 28-day case-fatality in the 65 year and older. AB - BACKGROUND AND OBJECTIVES: The myocardial infarction (MI) incidence rate, prognosis and hospitalisation rate in the population 65 and over are rarely studied. We sought to determine MI hospitalisation and incidence rates, and 28 day case-fatality, in the 65 year and older population, and to analyse whether their management and prognosis differed from that of younger patients. METHODS: All residents in Gerona (Spain) older than 24 years with suspected fatal or non fatal MI were investigated and included in a population registry. RESULTS: MI mortality, incidence, and case-fatality dramatically increased with age after 64. Smoking, thrombolysis, antiplatelet and betablocker drug use, coronary angiograms, and coronary revascularisation decreased with age. The risk of death of patients between 75 and 84 years (OR: 4.15, 95% confidence interval, CI: 1.70 10.15) and between 85 and 94 years (OR: 4.68, 95% CI: 1.62-13.52) was higher than in the 34-64 years age group, independently of any patient characteristic. CONCLUSIONS: The magnitude of the impact of MI in the elderly at population and hospital levels is substantially higher than in those younger than 65 years of age. After this age patients receive less treatments and procedures than their younger counterparts. PMID- 15117117 TI - Validity of self-reported information on cancer: determinants of under- and over reporting. AB - The first aim of the present study was to assess the validity of self-reported information concerning previous cancer. The second aim was to investigate determinants for under- and over-reporting of cancer. The Malmo Diet and Cancer Study is a prospective cohort study including 28,098 subjects. A questionnaire assessed lifestyle and medical conditions, e.g. previous cancer (yes/no). Cancer follow-up was made using the Swedish Cancer Registry (SCR). Questionnaire information concerning previous cancer was considered as test, and SCR information as golden standard (any tumour in the SCR, and specific sites were investigated). The validity analysis assessed agreement, sensitivity and specificity. Under- and over-reporting were studied in relation to lifestyle and socio-demographic factors. Sensitivity of the questionnaire in relation to malignant tumours was 0.82. It was 0.53 when cancer in situ of the breast and uterine cervix together with benign lesions of the urinary bladder and the nervous system were included in the cancer definition. Under- and over-reporting (in relation to malignant tumours) were more common among women and old people. Obesity, size of household, place of birth, smoking, social participation, educational level, type of employment, alcohol consumption and poor well-being was associated with under- or over-reporting in relation to at least one of the studied endpoints in either men or women. We conclude that there was a considerable disagreement between self-reported information and cancer registry data. PMID- 15117118 TI - Uterine fibroids risk and history of selected medical conditions linked with female hormones. AB - To understand the role of several medical conditions on the risk of uterine fibroids, we analysed the findings of a large case-control study. Cases were 843 women aged 54 or less (median age 43 years, range 21-54) with histologically confirmed uterine fibroids, whose clinical diagnosis dated back no more than 2 years. Indications for surgery were recurrent menorrhagia or ultrasound evidence of fibroids larger than 10 cm in diameter. Controls were 1557 women aged 54 years or less of comparable quinquennia of age (median age 43 years, range 21-54) who had not undergone hysterectomy and were admitted for acute, non-gynecologic, non hormonal, non-neoplastic conditions to a network of hospitals with a similar catchment area. Clinical history of severe overweight was inversely associated with the risk of fibroids (multivariate odds ratio (OR), OR: 0.6, 95% confidence intervals (CI): 0.5-0.8). An increase in the frequency of fibroids was seen in women with a history of benign breast disease (OR: 1.2, 95% CI: 0.9-1.6) and particularly of breast biopsies (OR: 2.0, 95% CI: 1.2-3.5). The results of this large dataset indicate that medical conditions known or likely to be related to female hormones are not important determinants of the risk of fibroids. PMID- 15117119 TI - Neonatal screening for glucose-6-phosphate dehydrogenase deficiency fails to detect heterozygote females. AB - We examined glucose-6-phosphate dehydrogenase (G6PD) deficiency in north-eastern Italian Caucasian neonates detected by neonatal screening, in order to measure the incidence of heterozygote females detected by neonatal screening, and to estimate the near-true total incidence. A total of 85,437 Caucasian neonates, born between January 2000 and December 2001, have been enclosed in the study. The total incidence of the disease, measured by fluorescent method, is 0.9 per thousand; the total incidence, calculated by Hardy-Weinberg law, is 4.8 per thousand. The frequency of missed females is 93% of total females expected with G6PD deficiency; most of them are very likely heterozygous females. The sensitivity of the fluorescent method might be not sufficient to detect all females. Since heterozygote females might develop the symptoms of G6PD deficiency later, these results suggest that the G6PD neonatal screening may not be helpful in preventing disease in females. PMID- 15117120 TI - Epidemiological evidence for vaccinating wastewater treatment plant workers against hepatitis A and hepatitis B virus. AB - Exposed to sewage workers of a wastewater treatment plant and a control group in Thessaloniki, Greece, were examined for antibodies against hepatitis A (anti-HAV) and hepatitis B virus (HBV) infection markers. The main objective of this study was to investigate for epidemiological evidence so as to recommend vaccination of the occupationally exposed workers against these viral infections. Antibodies against hepatitis A virus were detected in 65.7% of the wastewater treatment plant workers and in 32.6% of the control group. The prevalence of anti-HAV was significantly higher in less educated persons and was increasing with age (p < 0.001), whereas in logistic regression analysis the adjusted seroprevalence of wastewater treatment plant workers was 3.5 times higher (p < 0.01) than the control population. Serologic evidence of past HBV infection was observed in 32.4% of the exposed to sewage population and in 5.8% of the controls. Multivariate analysis showed that variables significantly and independently related to previous HBV infection was occupational exposure to sewage (OR: 5.81; 95% CI: 2.07-16.29) and age over 40 years old (OR: 4.49; 95% CI: 1.51-13.33). These results support the vaccination policy of young and sensitive to hepatitis A and B virus wastewater treatment plant workers in our region. PMID- 15117121 TI - Ocular involvement in cutaneous leishmaniasis four cases with blepharoconjunctivitis. AB - PURPOSE: To determine the rate of ocular involvement in cutaneous leishmaniasis (CL). METHODS: Nine hundred and eighty seven cutaneous leishmaniasis patients diagnosed and identified by clinical examination and parasitological methods in Harrankapi Health Centre between January 1996 and November 1997 were examined at the Harran University School of Medicine Eye Clinic, Sanliurfa, Turkey. RESULTS: One thousand seven hundred and three CL lesions were identified in 987 patients (484 women and 503 men) with ages ranging from 0 to 78 years (mean +/- SD 16.70 +/- 13.47 years). A total of 33 lesions were located on the eyelid and in the periorbital region, making up 1.93% of all cutaneous lesions and 3.57% of facial lesions. In three cases with ocular involvement, blepharoconjunctivitis was identified, and in one case with ocular involvement, mechanical ptosis, lagophthalmos and blepharoconjunctivitis were identified. CONCLUSIONS: It was observed that the size of the lesions and their vicinity to the marginal free edge of the eyelid played an important role in the occurrence of ocular symptoms. It was concluded that many ocular findings could be missed because of the failure to conduct ophthalmological examination of CL cases with eyelid involvement, and hence, proper follow up and treatment may not be carried out. PMID- 15117122 TI - The epidemiology of hepatitis a virus infection in children, in Edirne, Turkey. AB - Turkey is a middle endemic area with respect to Hepatitis A virus (HAV) infection. However, the frequency of this infection varies due to socio-economic differences in various regions. The aim of this study was to detect the most likely age of exposure to HAV and factors affecting infection rates among children living in Edirne. A sample of 645 children between the ages of 0-19 living in Edirne were tested for total anti-HAV levels using ELISA method. A questionnaire on socio-economic status (SES), possible risk factors, and place of residence was completed for each child. Anti-HAV seropositivity was found to be 4.4, 25, 37.3 and 43.2%, in 2-5, 6-10, 11-14 and 15-19 age groups, respectively. Seropositivity was found to be increasing with age (p < 0.05). According to logistic regression analysis results; mother's education, SES of family, history of hepatitis in primary family members and the number of brothers or sisters were determined as factors increasing the seropositivity of HAV. Furthermore, HAV infection risk was found to be a decreasing function of income that is higher the income less likely the infection. These results showed that HAV infection rate in Edirne is in middle endemicity and the most likely way of exposure is transmission from family members. Although it requires further cost-effectiveness studies, our results indicate that applying the HAV vaccination in early childhood would be beneficial to decrease the prevalence of the infection and prevent HAV epidemics. PMID- 15117124 TI - Prevalence of HIV, HTLV-I and HTLV-II among female sex workers in Spain, 2000 2001. AB - Using an unlinked anonymous survey the seroprevalence of HIV, HTLV-I and HTLV-II was analysed among female sex workers. They were surveyed when they attended sexually transmitted disease clinics in six Spanish cities during the period 2000 2001. Fifty-eight percent of the 3149 women analysed came from Latin America or sub-Saharan Africa. The total prevalence of HIV was 0.7%, rising to 15.9% amongst injecting drug users (IDUs). When this group was not included, the prevalence amongst the Latin-Americans or sub-Saharan Africans was 0.8% and amongst the women from other origins 0.3% (p = 0.148). 33.3% of the women infected with HIV already knew about their infection. The prevalences of HTLV-I and HTLV-II were 0.3% and 0.2%, respectively. The prevalences of these three infections in this collective were low if evaluated without the IDUs. PMID- 15117125 TI - Management of severe falciparum malaria in African children. PMID- 15117123 TI - Age-specific seroprevalence of hepatitis A infection among children visited in pediatric hospitals of Tehran, Iran. AB - BACKGROUND: Hepatitis A is an enterically transmitted disease that still remains endemic in many developing countries. In some countries improvements in living conditions have recently led to changing in epidemiology of hepatitis A virus (HAV) infection. In our country there are very few reports on prevalence of HAV infection. OBJECTIVE: To determine the seroprevalence of anti-HAV IgG among children visited in pediatric hospitals of Tehran, Iran. METHODS: The study group included 1018 children who were 6 months-14.9 years of age. These children were visited in four major pediatric hospitals of Tehran. The children were separated to three age groups: Group 1 (6 months-4.9 years; n = 469), Group 2 (5.0-9.9 years; n = 290), and Group 3 (10.0-14.9 years; n = 259). Serum anti-HAV IgG was tested with commercial ELISA kits. The data were tested for statistical significance with chi2 test. RESULTS: In all subjects, seroprevalence of hepatitis A was 22.3% (95% CI: 19.7, 24.9). There was no significant difference between genders (22.2% vs. 22.5% in males and females, respectively) and among age groups (Group I was 22.1% and Group 3 was 25.9; p > 0.05). CONCLUSIONS: In summary, it seems that HAV infection is not highly endemic at least in some urban areas of Iran. On the basis of this epidemiologic data, post exposure prophylaxis would be necessary for children and young adults, and hepatitis A vaccination strategy should be revised. PMID- 15117126 TI - The Medicines Crossing Borders Project--improving the quality of donations. PMID- 15117127 TI - Fluid management of severe falciparum malaria in African children. AB - Most African children with severe malaria who die do so on the day of admission as a result of the complications of falciparum malaria. We highlight the value of a rapid structured triage assessment to look for emergency signs that will prioritize initial management and implementation of basic life support. This can be delivered with few resources and by non-specialist medical personnel. Reduction in case fatality can only come through the wider appreciation of the need for and application of supportive therapies to treat the life-threatening complications. Hypovolaemia has emerged as a common feature of children presenting with severe malaria complicated by acidosis. Early recognition and prompt treatment may lead to improvements in outcome. We discuss the new evidence supporting the role of hypovolaemia in severe malaria and potential treatment options whilst awaiting the results of clinical trials. PMID- 15117128 TI - Management of seizures in children with falciparum malaria. AB - Falciparum malaria is the most common cause of convulsions in children admitted to hospital in malaria endemic areas. Detection of convulsions requires close monitoring and one should be on the look out for signs of subtle convulsions. About a third of acute seizures in children with cerebral malaria, do not manifest as convulsions, but as changes in eye deviation, salivation and/or eye deviation. It is important to terminate convulsions lasting more than 5 min, since prolonged convulsions are associated with neurological deficits in survivors of children with severe malaria. Initial management should include putting the child into the left lateral position, checking the blood glucose and administering oxygen if hypoxic. The benzodiazepines, particularly diazepam, are used as the initial anticonvulsants. Phenobarbital and phenytoin are used as second-line treatments. Prompt and effective management of falciparum malaria associated convulsions may contribute to a better outcome in children with severe malaria. PMID- 15117129 TI - Epidemiology of dog bites: a community-based study in India. AB - Rabies is a deadly disease with no cure which is mainly caused by dog bites. Data on dog bites and knowledge, attitude and practices (KAP) related to dog bites are not generally available to rural communities. We made a cross-sectional study of a rural community of ten villages served by a primary health centre selected by random sampling. Ten per cent of households selected by systematic random sampling were interviewed for KAP regarding dog bites. The dog bite rate was 25.7/1000 population per year. The rate for males was higher than for females; half of the bites were in summer; 40% did not go for any prophylaxis; half of the victims treated their wounds with chilli powder. Eighty per cent of the bites were class II or III and leg bites were most common. The majority of the community knew dog bites can cause death but they did not know the correct incubation period. Most knew that injections were available to prevent rabies. Only half said they would visit a hospital for treatment. Around 50% received tetanus toxoid. PMID- 15117130 TI - Traditional healers' practices and the spread of HIV/AIDS in south eastern Nigeria. AB - A 3-year (1997-1999) prospective study was carried out to evaluate the possible role of traditional healers (THs) in the spread of HIV/AIDS in south eastern Nigeria. Their patients' knowledge of HIV infection was also ascertained during the review period. Four contiguous states in south eastern Nigeria with high prevalent rates of HIV infection were chosen for the survey. Sixty-nine patients ofTHs were randomly selected and contacted from the patients' list of five selected THs in each state. Thus, a total of 20 THs and 69 patients formed subjects of the study. The study involved a face-to-face interview of the subjects by trained health personnel in each state using a prepared structured questionnaire. The questionnaire elicited information on their biodata, past/present medical history, social/family history, nature and mode of treatment received or administered, and their knowledge of HIV infection, where applicable. The results showed that there was a serious HIV/AIDS related risk inherent in the practices of Nigerian THs patronized by over 60% of the population. This was suggested by the continuous usage of unsterilized instruments and cross contamination with patients' blood and body fluid in their practices. It was further revealed that, the patients' HIV/AIDS related knowledge was poor (30%). Basic education on HIV/AIDS must be given to all groups in Nigeria including THs, their patients and general public. The World Health Organization and other agencies involved in the fight against HIV infection should focus on the safety of THs' practices as a possible mode of transmission of HIV infection in Africa. Unless this aspect is taken seriously along with other preventive strategies already adopted, the epidemic of the disease condition may continue to spread in Africa. PMID- 15117131 TI - Buruli ulcer in the Democratic Republic of Congo: epidemiology, presentation and outcome. PMID- 15117132 TI - Primary healthcare workers' perceptions about barriers to health services in Zambia. PMID- 15117133 TI - Community perception and readiness for anti-helminth programmes in rural Nepal. AB - We report on the community perception of helminth infection and readiness to participate in and pay for a community-based anti-helminth programme in rural Nepal. For data collection, 28 focus group discussions (FGDs) were conducted in 28 communities. The FGD results revealed that community members were lacking in scientific knowledge about helminth infection. They also had an unusual perception about it--they believed that a 'mul juka' (vital worm) should reside in every person's stomach from birth, and must never be killed. Nonetheless, the results showed that the community members were willing to participate in and were ready to contribute their resources to the anti-helminth programmes as long as modern medicine would not kill the 'mul juka'. Such community readiness provides us with hope for developing a sustainable anti-helminth programme at the community level through effective health education. PMID- 15117135 TI - Retraction of Sama MT, Homeida M, Ngang P, Liese BH, Amazingo U, Seketeli A. A multi-centre study of community-directed ivermectin distributors' (CDDs') involvement in other healthcare and development programme activities in Cameroon, Togo, Sudan, Nigeria and Uganda (Tropical Doctor 2003; 33: 237-241). PMID- 15117134 TI - Knowledge, attitudes and practices towards malaria among primary school pupils in Ndirande, Blantyre, Malawi. AB - A qualitative cross-sectional study was conducted at Ndirande Primary School in the peri-urban area of Blantyre, Malawi in 2002. The aim was to describe the knowledge, attitudes and practices towards malaria among pupils. Forty pupils (20 boys and 20 girls) were recruited into four focus group discussions. There was very good knowledge about the role of the female Anopheles mosquito as an agent for transmission of malaria. Respondents were also aware of the clinical features of malaria and what measures could be taken to prevent the disease. However, the survey showed that many of the environmental modification measures were difficult to implement in the overcrowded peri-urban areas. Respondents did not believe that they could influence parents or other authorities in instituting environmental modification interventions. We conclude that children are unlikely to be effective change agents if the social environment does not offer them the opportunity to influence key community figures. PMID- 15117136 TI - An accurate and portable system for glycated haemoglobin measurement in the tropics. AB - Measurement of glycated haemoglobin (HbA1c) is vital to provide meaningful diabetic care, but the assay is difficult and expensive, making its availability limited in resource-poor countries. We have field-tested a novel near-patient HbA1c meter (Glycosal; Provalis Diagnostics Ltd, UK) in northern Ethiopia. The machine was easy to use and gave results which correlated well (r = -0.96) with standard laboratory methods. The meter also performed well and retained accuracy at high ambient temperature (34.0 degrees C). Though still relatively expensive (pound 4 per test), this meter does give the opportunity for practical and appropriate HbA1c testing in tropical climes, and should be considered for at least intermittent use in diabetic patients. PMID- 15117137 TI - Atraumatic sutures can be made locally. AB - The usefulness of fishing line in surgical practice has previously been described in this journal. Now, this usefulness is further enhanced by the possibility of producing an atraumatic suture using a pair of modified pliers. PMID- 15117138 TI - Post-partum practices among women in Calabar, Nigeria. AB - We carried out a study on post-partum practices amongst women in Calabar at the maternal and child health clinic at University of Calabar Teaching Hospital between 1 March and 31 May 2002. Three hundred women were interviewed. One hundred and ninety-three (64.3%) resumed coitus prior to 6 weeks of puerperium. There was no significant difference in parity (P = 0.20) and educational status (P = 0.10) between women who resumed coitus earlier than 6 weeks and those who resumed later. One out of every 10 women resumed coitus without any contraception. The majority [212 (70.7%)] sat in hot water (sitz bath) to aid perineal wound healing and improve vaginal tone. PMID- 15117139 TI - Prevalence of hepatitis B surface antigen (HBsAG) in pregnant women in South Kanara District, Karnataka State, India. PMID- 15117140 TI - Availability of donor corneal tissue for transplantation. AB - The utilization rate of available donor corneas is very low almost everywhere. The present work was undertaken to study the rate of acceptability of donor corneas in a city eye bank in India. Donor eyes were evaluated under slit lamp examination and tissue quality was categorized as: very good; good; fair; and poor following the guidelines laid down by the Eye Bank Association of America in 1992. One hundred and eight eyes (from 54 donors) were studied. The mean age of the donors was 75.9 years (range 30-97 years). Thirty-nine (36%) eyes were of very good to good quality and of these 37 (34.3%) were acceptable for grafting either for optical or therapeutic purposes. Sixty-one (56.4%) eyes were not acceptable owing to fair to poor tissue quality. Four (3.7%) eyes were discarded on medical grounds and six (5.6%) were discarded both because of fair to poor tissue quality and on medical grounds. Twenty-three of 50 tissues (46%) available from donors aged 75 years or younger were acceptable while 14 of 58 (24.1%) tissues from donors over 75 years were acceptable. PMID- 15117141 TI - Helicobacter pylori eradication: efficacy of conventional therapy in India. PMID- 15117143 TI - Adhesion intestinal obstruction in children in northern Nigeria. AB - In developing countries, reports on adhesion intestinal obstruction in children are scanty. We report 30 children managed for adhesion intestinal obstruction during a 16-year period. The age range was 10 weeks-14 years (median 9 years). There were 24 boys and 6 girls. Postoperative adhesion was the cause in 13 (43%) patients, inflammatory in 11 (37%), and in 5 (17%) no cause could be identified. In one patient, adhesion followed missed ileal perforation from blunt abdominal trauma. Duration of symptoms was 1-21 days (median 4 days). Only four of the 13 patients with postoperative adhesion obstruction were managed conservatively initially, but this failed in all; one was found to have an intestinal perforation at laparotomy. The remaining nine had immediate laparotomy owing to presence of features of strangulation at presentation; two required intestinal resection for gangrene. All other patients had laparotomy soon after resuscitation. The resection rate for gangrene was 13% and 3% required closure of perforations. Postoperatively six (20%) patients developed eight infective complications. There was recurrence in three of 27 (11%) surviving patients within 3 months. Mortality was three (10%). The presentation of adhesion intestinal obstruction in children in northern Nigeria is late and morbidity and mortality are high. Early presentation should improve the outcome. PMID- 15117142 TI - Bacteriological profile in chronic suppurative otitis media in Eastern Nepal. AB - Aural swabs were collected from 250 patients with chronic suppurative otitis media (CSOM) (tubotympanic disease). Duration of ear discharge was more than 6 months for culture and sensitivity test. The disease was unilateral in 178 (71.2%) and bilateral in 72 (28.8%) patients. From a total of 322 aural swab cultures, 324 organisms were isolated with predominance of Pseudomonas aerugimosa (36.4%) followed by Staphylococcus aureus (30.2%). All the isolated Gram-negative organisms showed in vitro sensitivity to ciprofloxacin, amikacin, ofloxacin, and gentamycin whereas cloxacillin, ofloxacin, erythromycin, ciprofloxacin, gentamicin and tetracycline were more effective against Gram-positive isolates. Most isolates were resistant to amoxycillin. It is concluded that ciprofloxacin is preferred to aminoglycosides as an important tool in the treatment of CSOM because of its lower cost, lack of ototoxicity and availability as topical oral preparations. PMID- 15117144 TI - CA 19-9 in various gastrointestinal malignancies in an Indian population. PMID- 15117145 TI - Umbilical hernia coexisting with mobile caecum in Nigerian children. PMID- 15117146 TI - Hysterectomies amongst teenagers in Calabar, Nigeria. AB - A 10-year retrospective study of hysterectomies performed on teenagers in Calabar was carried out. PMID- 15117147 TI - Recurrence of multicystic ameloblastoma in soft tissue. AB - Ameloblastoma is characteristically considered to be a benign but locally aggressive and infiltrative odontogenic tumour with a high recurrence rate. Most recurrences of the solid or multicystic ameloblastoma occurred at the edges of the initial tumour or at the bony cavity left after the excision of a tumour. A case of an isolated soft tissue recurrence of ameloblastoma of the mandible in a 58-year-old man is presented. The report illustrates the need to take into consideration the adjacent soft tissue in cases of tumour with cortical bone perforation during surgical treatment. This is especially important in an environment where patients are not likely to attend follow-up review for a long period. It also demonstrates the ability of intraosseous ameloblastoma to grow in soft tissues. PMID- 15117148 TI - Periorbital infection in the infant. PMID- 15117149 TI - Urinary trichomoniasis among sexually active Thai females. PMID- 15117151 TI - Age-specific prevalence of hepatitis B infection among the Karen in the Andaman and Nicobar Islands, India. PMID- 15117150 TI - Conflict and home birth increase in the West Bank. PMID- 15117152 TI - Utilization of hearing aids by the hearing-impaired in Ghana. PMID- 15117153 TI - Spectrum of tuberculosis in HIV-positive patients in Uttaranchal, India. PMID- 15117154 TI - Penicillin-resistant Streptococcus pneumoniae at a tertiary care centre in Pakistan. PMID- 15117155 TI - A preliminary study on extended-spectrum beta-lactamase producing Klebsiella species from a South Indian hospital. PMID- 15117156 TI - Pericardial effusion and tamponade in Zaria: the association of HIV/AIDS. PMID- 15117157 TI - Suicide in Nepal: facts and figures. PMID- 15117158 TI - A psychosocial intervention programme for malnourished Haitian children. PMID- 15117159 TI - Absence of antibody to HTLV-1 in sera of Asian Indian patients with systemic autoimmune rheumatic diseases. PMID- 15117160 TI - Cancer registration in a developing country: problems and prospects of the new UDUTH Cancer Registry, Sokoto, Nigeria. PMID- 15117161 TI - Trapped head after fetal demise. PMID- 15117162 TI - Reasons for non-compliance with antihypertensive medication in southwest Nigeria. PMID- 15117163 TI - Birds, viruses, and history: the current 'genuine adventure'. PMID- 15117164 TI - Hyaluronic acid minimally effective for knee degenerative joint disease. PMID- 15117165 TI - Avian influenza: a wake-up call from birds to humans. PMID- 15117166 TI - For low platelets, how low is dangerous? PMID- 15117167 TI - Chest pain relief by nitroglycerin doesn't predict active coronary artery disease. PMID- 15117168 TI - Major bleeding risk with warfarin for deep vein thrombosis. PMID- 15117169 TI - Antioxidant studies need a change of direction. PMID- 15117170 TI - Minimally invasive surgery for morbid obesity. AB - In patients with morbid obesity, surgery is the only treatment known to produce sustained weight loss and to reduce comorbidities. Traditional (open) gastric bypass surgery is associated with postoperative complications that include wound infection, pulmonary embolism, and pneumonia. Laparoscopic techniques achieve similar long-term results as does open surgery, with fewer postoperative complications. PMID- 15117171 TI - Premenstrual dysphoric disorder: a review for the treating practitioner. AB - Premenstrual dysphoric disorder (PMDD), a severe form of premenstrual syndrome (PMS), is characterized by physical and behavioral symptoms that cause marked social impairment during the last half of the menstrual cycle. Symptoms are believed to result from the interaction of central neurotransmitters and normal menstrual hormonal changes. Treatment usually begins with lifestyle changes, over the-counter medications, and if needed, selective serotonin reuptake inhibitors. Physicians should be aware of the risks of many of the alternative therapies commonly touted in the popular press. PMID- 15117172 TI - Patient information. What is premenstrual dysphoric disorder (PMDD)? PMID- 15117173 TI - Tricyclics, not SSRIs, effective for chronic back pain. PMID- 15117174 TI - Recent trials of antioxidant therapy: what should we be telling our patients? AB - The current evidence does not support the indiscriminate use of vitamins A, C, or E or beta carotene to prevent or reduce cardiovascular disease. Despite a plausible theory that antioxidants can prevent diseases caused by oxidative damage, trials thus far have not proven this. In fact, some studies found antioxidants may be harmful in some people. We review important studies of the effects of four antioxidants (vitamins A, C, and E, and beta carotene) and analyze whether the current evidence supports or confirms or rejects the presumed protective role. PMID- 15117175 TI - Hypoglycemia in diabetes: common, often unrecognized. AB - Hypoglycemic episodes in patients with diabetes often go unrecognized, and over time, patients may lose the ability to sense hypoglycemia, increasing their risk. Intensive diabetes control is beneficial for patients with diabetes, but it increases their risk of hypoglycemia, underscoring the complexity of diabetes management. PMID- 15117176 TI - Patient information. Hypoglycemia. PMID- 15117177 TI - Perioperative nutrition support: who and how. AB - Perioperative nutrition support can reduce postoperative complications in some malnourished patients, but there are risks, such as a greater risk of infection. The decision to use nutrition support--either total parenteral nutrition or enteral feedings--before and after surgery depends on how severely the patient is malnourished, the type of surgical procedure, and whether the surgery is elective. PMID- 15117178 TI - Head injury: which patients need imaging? Which test is best? AB - Some patients with head injuries definitely need to undergo an imaging study- usually computed tomography (CT). Most, however, are in a category of "apparently mild" injury, and controversy continues about which of them need to undergo imaging studies to rule out intracranial injuries. PMID- 15117179 TI - Multistep microreactions with proteins using electrocapture technology. AB - A method to perform multistep reactions by means of electroimmobilization of a target molecule in a microflow stream is presented. A target protein is captured by the opposing effects between the hydrodynamic and electric forces, after which another medium is injected into the system. The second medium carries enzymes or other reagents, which are brought into contact with the target protein and react. The immobilization is reversed by disconnecting the electric field, upon which products are collected at the outlet of the device for analysis. On-line reduction, alkylation, and trypsin digestion of proteins is demonstrated and was monitored by MALDI mass spectrometry. PMID- 15117180 TI - Factors affecting quantitative analysis in laser desorption/laser ionization mass spectrometry. AB - Microprobe laser desorption/laser ionization mass spectrometry (microL(2)MS) is a sensitive and selective technique that has proven useful in the qualitative and semiquantitative detection of trace organic compounds, particularly polycyclic aromatic hydrocarbons (PAHs). Recent efforts have focused on developing microL(2)MS as a quantitative method, often by measuring the ratio of signal strength of an analyte to an internal standard. Here, we present evidence of factors that affect these ratios and thus create uncertainty and irreproducibility in quantification. The power and wavelength of the desorption laser, the delay time between the desorption and ionization steps, the power of the ionization laser, and the ionization laser alignment are all shown to change PAH ratios, in some cases by up to a factor of 24. Although changes in the desorption laser parameters and the delay time cause the largest effects, the ionization laser power and alignment are the most difficult parameters to control and thus provide the most practical limitations for quantitative microL(2)MS. Variation in ratios is seen in both synthetic poly(vinyl chloride) membranes and in "real-life" samples of Murchison meteorite powder. Ratios between similar PAHs vary less than those between PAHs that differ greatly in mass and structure. This finding indicates that multiple internal standards may be needed for quantification of samples containing diverse PAHs. PMID- 15117181 TI - Top-down characterization of nucleic acids modified by structural probes using high-resolution tandem mass spectrometry and automated data interpretation. AB - A top-down approach based on sustained off-resonance irradiation collision induced dissociation (SORI-CID) has been implemented on an electrospray ionization (ESI) Fourier transform mass spectrometer (FTMS) to characterize nucleic acid substrates modified by structural probes. Solvent accessibility reagents, such as dimethyl sulfate (DMS), 1-cyclohexyl-3-(2 morpholinoethyl)carbodiimide metho-p-toluenesulfonate (CMCT), and beta-ethoxy alpha-ketobutyraldehyde (kethoxal, KT) are widely employed to reveal the position of single- vs double-stranded regions and obtain the footprint of bound proteins onto nucleic acids structures. Established methods require end-labeling of the nucleic acid constructs, probe-specific chemistry to produce strand cleavage at the modified nucleotides, and analysis by polyacrylamide gel electrophoresis to determine the position of the susceptible sites. However, these labor-intensive procedures can be avoided when mass spectrometry is used to identify the probe induced modifications from their characteristic mass signatures. In particular, ESI-FTMS can be directly employed to monitor the conditions of probe application to avoid excessive alkylation, which could induce unwanted distortion or defolding of the substrate of interest. The sequence position of the covalent modifications can be subsequently obtained from classic tandem techniques, which allow for the analysis of individual target adducts present in complex reaction mixtures with no need for separation techniques. Selection and activation by SORI CID has been employed to reveal the position of adducts in nucleic acid substrates in excess of 6 kDa. The stability of the different covalent modifications under SORI-CID conditions was investigated. Multiple stages of isolation and activation were employed in MS(n)() experiments to obtain the desired sequence information whenever the adduct stability was not particularly favorable, and SORI-CID induced the facile loss of the modified base. A new program called MS2Links was developed for the automated reduction and interpretation of fragmentation data obtained from modified nucleic acids. Based on an algorithm that searches for plausible isotopic patterns, the data reduction module is capable of discriminating legitimate signals from noise spikes of comparable intensity. The fragment identification module calculates the monoisotopic mass of ion products expected from a certain sequence and user defined covalent modifications, which are finally matched with the signals selected by the data reduction program. Considering that MS2Links can generate similar fragment libraries for peptides and their covalent conjugates with other peptides or nucleic acids, this program provides an integrated platform for the structural investigation of protein-nucleic acid complexes based on cross-linking strategies and top-down ESI-FTMS. PMID- 15117182 TI - An operator-independent approach to mass spectral peak identification and integration. AB - A mathematical algorithm is presented that locates and calculates the area beneath peaks from real data using only reproducible mathematical operations and no user-selected parameters. It makes no assumptions about peak shape and requires no smoothing or preprocessing of the data. In fact, it is shown that for matrix-assisted laser desorption time-of-flight mass spectra noise exists at all frequency ranges making the smoothing of data without distortion of peak areas impossible. The algorithm is based on a time-series segmentation routine that reduces the data set to groups of three strategic points where each group defines the beginning, center, and ending of each peak located. The peak areas are found from the strategic points using a commonplace polygonal area calculation routine. Peaks with statistically insignificant height or area are then discarded. The performance of the algorithm is demonstrated on a polystyrene mass spectrum with varying degrees of noise added either mathematically or experimentally. An on line implementation of the method, termed MassSpectator, for public use can be found at www.nist.gov/maldi. PMID- 15117183 TI - A liquid chromatography--tandem mass spectrometry multiresidue method for quantification of specific metabolites of organophosphorus pesticides, synthetic pyrethroids, selected herbicides, and deet in human urine. AB - The ability to estimate low-dose human exposure to commonly used pesticides often is requested in epidemiologic studies. Therefore, fast and robust methods are necessary that can measure many analytes in the same sample. We have developed a method for high-throughput analysis of 19 markers of commonly used pesticides in human urine. The analytes were seven specific metabolites of organophosphorus pesticides, five metabolites of synthetic pyrethroids, six herbicides or their metabolites, and one insect repellant. Human urine (2 mL) was spiked with stable isotopically labeled analogues of the analytes, enzymatically hydrolyzed, extracted using solid-phase extraction, concentrated, and analyzed using high performance liquid chromatography-tandem mass spectrometry. The sample was divided into two portions and analyzed on two different mass spectrometers, one using atmospheric pressure chemical ionization (APCI) and the other using turbo ion spray atmospheric pressure ionization (TIS). All analytes except the pyrethroid metabolites were analyzed using APCI. The detection limits for all analytes ranged from 0.1 to 1.5 ng/mL of urine, with the majority (17) below 0.5 ng/mL. The analytical precision for the different analytes, estimated as both the within-day and between-day variation, was 3-14 and 4-19%, respectively. The extraction recoveries of the analytes ranged from 68 to 114%. The throughput, including calibration standards and quality control samples, is approximately 50 samples a day. However, the analysis time with the TIS application is much shorter, and if only pyrethroid metabolite data are of interest, the throughput can be increased to 100-150 samples/day. PMID- 15117184 TI - Atmospheric pressure MALDI with pulsed dynamic focusing for high-efficiency transmission of ions into a mass spectrometer. AB - The atmospheric pressure (AP) matrix-assisted laser desorption/ionization (MALDI) technique described to date has proven to be a convenient and rapid method for soft ionization of biomolecules. However, this technique, like other AP ionization methods, has so far suffered from a low efficiency in transmitting ions from atmospheric pressure into the vacuum of the mass spectrometer (MS). In this work, a novel technique we termed pulsed dynamic focusing, or PDF, which improves the ion transmission efficiency and sensitivity of AP-MALDI by over an order of magnitude, is described. Pulsed dynamic focusing operates on the basis of pulsing a high-voltage extraction field to zero, when ions are just outside of the MS entrance, to allow the intake gas flow of the MS to effectively entrain the ions into the MS. Results from application of the PDF technique to an AP MALDI ion trap MS demonstrated that in comparison to static AP-MALDI operation (1). up to 2.1 times more ions from a given laser shot could be transferred into the MS, (2). applying higher voltages in combination with the switching scheme yielded up to 1.6-times-higher ion intensities, and (3). a 3-times-larger laser spot area could be utilized. The combination of these factors produced an enhancement in throughput and sensitivity, as measured by the ions detected per unit time, of over 12 times for a digest sample of bovine serum albumin. In addition, the PDF technique proved to make AP-MALDI less sensitive to laser positioning, creating a more robust ion source in comparison to static AP-MALDI. PMID- 15117185 TI - Tuning sensitivity and selectivity of complementary metal oxide semiconductor based capacitive chemical microsensors. AB - New details on selectivity and sensitivity of fully integrated CMOS-based capacitive chemical microsensor systems are revealed. These microsystems have been developed to detect volatile organics in ambient air and rely on polymeric sensitive layers. The sensitivity and selectivity changes induced by thickness variation of the sensitive polymer layer allow for tuning of the layer parameters to achieve desired sensor features. Cross-sensitivity to interfering agents can be drastically reduced, as is shown for two important cases: (a). rendering the capacitive sensor insensitive to a low-dielectric-constant analyte (lower than that of the polymer) and (b). reducing the influence of a high-dielectric constant analyte, such as water, on the sensor response. The second case is of vital importance for capacitive sensors, since water is omnipresent and evokes large capacitive sensor signals. The thickness-induced selectivity is explained as a combination of dielectric constant change and swelling and has been confirmed by measurements. Experimentally determined sensitivities qualitatively and quantitatively coincide with the calculated values implying understanding of the sensing mechanism. PMID- 15117186 TI - Detection of femtomolar concentrations of HF Using an SiO(2) microcantilever. AB - Femtomolar concentrations of hydrogen fluoride, a decomposition component of nerve agents, were detected using a SiO(2) microcantilever. The microcantilever underwent bending due to the reaction of HF with SiO(2). The microcantilever deflection increased as the concentration of HF increased. Other acids, such as HCl, had no effect on the deflection of the cantilever. The mechanism of reaction induced bending and the correlation of microcantilever deflection with the HF concentration are discussed. The deflection in response to HF of a commercially available silicon cantilever was also studied, and its response was compared with that of the SiO(2) cantilever. Much less bending amplitude and sensitivity were observed for the silicon cantilever. PMID- 15117187 TI - Development of a microfabricated palladium decoupler/electrochemical detector for microchip capillary electrophoresis using a hybrid glass/poly(dimethylsiloxane) device. AB - The fabrication and evaluation of a palladium decoupler and working electrode for microchip capillary electrophoresis (CE) with electrochemical detection is described. The use of the Pd decoupler allows the working electrode to be placed directly in the separation channel and eliminates the band-broadening characteristic of the end-channel configuration. The method used for fabrication of the decoupler and working electrode was based on thin-layer deposition of titanium followed by palladium onto a glass substrate. When employed as the cathode in CE, palladium absorbs the hydrogen gas that is generated by the hydrolysis of water. The effect of the decoupler size on the ability to remove hydrogen was evaluated with regard to reproducibility and longevity. Using boric acid and TES buffer systems, 500 microm was determined to be the optimum decoupler size, with effective voltage isolation lasting for approximately 6 h at a constant field strength of 600 V/cm. The effect of distance between the decoupler and working electrode on noise and resolution for the separation of dopamine and epinephrine was also investigated. It was found that 250 microm was the optimum spacing between the decoupler and working electrode. At this spacing, laser-induced fluorescence detection at various points around the decoupler established that the band broadening due to pressure-induced flow that occurs after the decoupler did not significantly affect the separation efficiency of fluorescein. Limits of detection, sensitivity, and linearity for dopamine (500 nM, 3.5 pA/microM, r(2) = 0.9996) and epinephrine (2.1 microM, 2.6 pA/microM, r(2) = 0.9996) were obtained using the palladium decoupler in combination with a Pd working electrode. PMID- 15117188 TI - Direct manipulation and observation of the rotational motion of single optically trapped microparticles and biological cells in microvortices. AB - This paper describes a method for manipulating and monitoring the rotational motion of single, optically trapped microparticles and living cells in a microvortex. To induce rotation, we placed the microparticle at the center of rotation of the vortex and used the recirculating fluid flow to drive rotation. We have monitored the rotation of single beads (which ranged in diameter from a few micrometers to tens of micrometers) and living cells in a microvortex. To follow the rotation of a smooth and symmetrically shaped bead, we first ablated a small region ( approximately 1 microm) on the bead. An Ar(+) laser was then tightly focused ( approximately 0.5-microm spot size) onto the bead, and rotation was tracked by recording changes in the level of backscattered laser light as the ablated region repeatedly transited the laser focus. Using this method, we have followed bead rotation that varied in frequency from 0.15 to 100 Hz and have studied the effect of bead diameter on the rate of rotation at a given fluid flow rate. To monitor the rotation of single living cells, we selectively stained portions of B-lymphocytes with the fluorescent dye DiOC(6). We observed rotation by following changes in the fluorescence signal as the dye-stained region transited the laser focal volume. This technique provides a simple and sensitive method for controlling and monitoring the rotational motion of microparticles in a microfluidic environment. PMID- 15117189 TI - Real-time measurements of dissolved oxygen inside live cells by organically modified silicate fluorescent nanosensors. AB - Optical PEBBLE (probes encapsulated by biologically localized embedding) nanosensors have been developed for dissolved oxygen using organically modified silicate (ormosil) nanoparticles as a matrix. The ormosil nanoparticles are prepared via a sol-gel-based process, which includes the formation of core particles with phenyltrimethoxysilane as a precursor followed by the formation of a coating layer with methyltrimethoxysilane as a precursor. The average diameter of the resultant particles is 120 nm. These sensors incorporate the oxygen sensitive platinum porphyrin dye as an indicator and an oxygen-insensitive dye as a reference for ratiometric intensity measurement. Two pairs of indicator dye and reference dye, respectively, platinum(II) octaethylporphine and 3,3' dioctadecyloxacarbocyanine perchlorate, and platinum(II) octaethylporphine ketone and octaethylporphine, were used. The sensors have excellent sensitivity with an overall quenching response of 97%, as well as excellent linearity of the Stern Volmer plot (r(2) = 0.999) over the whole range of dissolved oxygen concentrations (0-43 ppm). In vitro intracellular changes of dissolved oxygen due to cell respiration were monitored, with gene gun injected PEBBLEs, in rat C6 glioma cells. A significant change was observed with a fluorescence ratio increase of up to 500% after 1 h, for nine different sets of cells, which corresponds to a 90% reduction in terms of dissolved oxygen concentration. These results clearly show the validity of the delivery method for intracellular studies of PEBBLE sensors, as well as the high sensitivity, which is needed to achieve real-time measurements of intracellular dissolved oxygen concentration. PMID- 15117190 TI - Detection of glutamate in optically trapped single nerve terminals by Raman spectroscopy. AB - Glutamate is the major excitatory neurotransmitter and is of particular interest in light of current models of memory and learning. The paper describes the first in situ detection of glutamate in single nerve terminals (synaptosomes), which is achieved by using laser trapping Raman spectroscopy. The near-infrared laser light captures a single synaptosome obtained from a Wister rat brain. The release of glutamate in a single laser-trapped synaptosome was detected by subtracting the Raman spectrum before depolarization from that after depolarization with the addition of the K(+)-channel blocker, 4-aminopyridine. The result indicated that the single synaptosome released approximately 3 amol of glutamate and that the release rate depended on the 4-aminopyridine concentration. PMID- 15117191 TI - Two- and three-dimensional van krevelen diagrams: a graphical analysis complementary to the kendrick mass plot for sorting elemental compositions of complex organic mixtures based on ultrahigh-resolution broadband fourier transform ion cyclotron resonance mass measurements. AB - Ultrahigh-resolution electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry has resolved and identified the elemental compositions of over 10000 organic constituents of coal and petroleum crude oil. A plot of Kendrick mass defect versus Kendrick nominal mass sorts compounds into homologous series according to compound class (i.e., numbers of N, O, and S heteroatoms), type (number of rings plus double bonds), and degree of alkylation (number of CH(2) groups), to yield unique elemental assignments from ultrahigh resolution mass measurements in the 200-900 Da range. Interpretation of such a vast compilation requires a simple (preferably graphical) means to differentiate between complex organic mixtures of different origin or processing. In an extension of the recently revived van Krevelen plot, each elemental composition is projected onto two or three axes according to its H/C, O/C, and/or N/C atomic ratios. The H/C ratio separates compounds according to degree of saturation, whereas O/C or N/C ratios separate according to O and N classes. We show that the three-dimensional van Krevelen diagram can completely separate different classes in pyridine-extracted coal or petroleum samples and can also graphically distinguish fossil fuels according to their nature (coal vs petroleum), maturation (coals of different rank), and processing (the same coal at two stages of liquefaction). The van Krevelen diagram thus appears well suited to amplifying and exposing compositional differences within and between complex organic mixtures. PMID- 15117192 TI - Real-time, on-line characterization of diesel generator air toxic emissions by resonance-enhanced multiphoton ionization time-of-flight mass spectrometry. AB - The laser-based resonance-enhanced multiphoton ionization time-of-flight mass spectrometry (REMPI-TOFMS) technique has been applied to the exhaust gas stream of a diesel generator to measure, in real time, concentration levels of aromatic air toxics. Volatile organic compounds, as well as several polycyclic aromatic hydrocarbons were detected in the concentration range of 10-200 ppb in the steady state diesel generator exhaust. The results were verified and compared with conventional extractive sampling and analytical techniques using gas chromatography/mass spectrometry (GC/MS). The high isomer selectivity of the REMPI-TOFMS instrument provided data for individual xylene isomers that are otherwise (partially) coeluting in standard GC/MS analyses. Good agreement was observed between results for volatile and semivolatile organic compounds obtained with REMPI-TOFMS and conventional extractive sampling. Transient events, such as cold start-ups of the diesel generator, resulted in sharp (less than 15 s) peak emissions that were, for benzene, up to a factor of 90 higher than the predominately constant concentrations observed during steady-state operation; warm restarts resulted in lower peak concentrations by a factor of 2.5. These fast transient emissions are only detectable using a real-time approach (1-s resolution) as demonstrated here using REMPI-TOFMS. PMID- 15117193 TI - Comprehensive two-dimensional normal-phase (adsorption)-reversed-phase liquid chromatography. AB - A comprehensive two-dimensional HPLC system has been developed. It is based on the use of a microbore silica column operated in normal-phase (adsorption) mode (NP) in the first dimension and a monolithic type C18 column operated in reversed phase (RP) mode in the second dimension. The interface was a 10-port, 2-position valve equipped with two storage loops. The first column was operated at a flow rate of 20 microL/min in isocratic mode, while the monolithic column flow rate was 4 mL/min and was operated in gradient mode. The sample loops had a volume of 20 microL each, and the analysis time in the second dimension was 1 min. In this way, every fraction from the first dimension was transferred on-line to the second dimension switching the automated valve every minute. A photodiode array detector has been used after the secondary column. The use of normal- and reversed-phase mode in the two dimensions can be helpful in the separation of complex mixtures of a natural origin that contain uncharged molecules of comparable dimension, different in polarity and hydrophobicity. The use of a microbore column in the first dimension permits the injection of a small volume in the secondary column, making the transfer of incompatible solvents from the first to the second dimension possible. Since the mobile phase in the NP separation is always stronger than the mobile phase at the head of the secondary column operated in RP mode, the initial eluent strength is important in order to obtain an effective focusing of the sample. The use of a monolithic type column in the second dimension permits the performance of very fast analysis operating at higher flow rates without loss of resolution, due to a higher permeability and increased mass-transfer properties in comparison to conventional particulate columns. Due to the brief reconditioning time necessary for monolithic columns, repetitive gradients can be carried out, extending the field of application to mixtures that contain components with different polarities. The utility of the system has been demonstrated in the analysis of the oxygen heterocyclic fraction of cold-pressed lemon oil, made up of coumarins and psoralens. These components may contain hydroxyl, methoxyl, isopentenyl, isopentenyloxyl, and geranyloxyl groups and oxygen-containing modification of the terpenoid side-chain groups, such as epoxides or vicinal diol groups. The relative location of the components in the 2D plane varied in relation to their chemical structure and allowed positive peak identification. The UV spectra recorded with the photodiode array detector supplied additional information that was used for the characterization of the studied sample. PMID- 15117194 TI - Characterization of a focal plane camera fitted to a Mattauch-Herzog geometry mass spectrograph. 2. Use with an inductively coupled plasma. AB - A novel charge-sensitive detector array, termed the focal plane camera (FPC), has been coupled to a Mattauch-Herzog mass spectrograph (MHMS) with an inductively coupled plasma ionization source. The FPC employs an array of gold Faraday cups, each with its own charge-integrating circuit that allows the simultaneous detection of several m/z ratios. The ion-sampling interface of the MHMS has been redesigned to provide better heat transfer away from the sampler and skimmer cones and to reduce the negative effects of turbulent gas flows around the plasma. The instrument has produced limits of detection in the tens to hundreds of parts per quadrillion regime and isotope ratio accuracy and precision of 5% error and 0.007% RSD, respectively. Limits of detection with the FPC are comparable to those obtained with a single-channel secondary electron multiplier (SEM). However, the isotope ratio accuracy and precision are better with the FPC than when the SEM is employed. The dynamic range has been shown to be linear over 7 orders of magnitude. PMID- 15117195 TI - Mechanistic studies on enzymatic reactions by electrospray ionization MS using a capillary mixer with adjustable reaction chamber volume for time-resolved measurements. AB - Mass spectrometry (MS)-based techniques have enormous potential for kinetic studies on enzyme-catalyzed processes. In particular, the use of electrospray ionization (ESI) MS for steady-state measurements is well established. However, there are very few reports of MS-based studies in the pre-steady-state regime, because it is difficult to achieve the time resolution required for this type of experiment. We have recently developed a capillary mixer with adjustable reaction chamber volume for kinetic studies by ESI-MS with millisecond time resolution (Wilson, D. J.; Konermann, L. Anal. Chem. 2003, 75, 6408-6414). Data can be acquired in kinetic mode, where the concentrations of selected reactive species are monitored as a function of time, or in spectral mode, where entire mass spectra are obtained for selected reaction times. Here, we describe the application of this technique to study the kinetics of enzyme reactions. The hydrolysis of p-nitrophenyl acetate by chymotrypsin was chosen as a simple chromophoric model system. On-line addition of a "makeup solvent" immediately prior to ionization allowed the pre-steady-state accumulation of acetylated chymotrypsin to be monitored. The rate constant for acetylation, as well as the dissociation constant of the enzyme-substrate complex obtained from these data, is in excellent agreement with results obtained by conventional stopped-flow methods. Bradykinin was chosen to illustrate the performance of the ESI-MS-based method with a nonchromophoric substrate. In this case, the unfavorable rate constant ratio for acylation and deacylation of the enzyme precluded measurements in the pre-steady-state regime. Steady-state experiments were carried out to determine the turnover number and the Michaelis constant for bradykinin. The methodologies used in this work open a wide range of possibilities for future ESI MS-based kinetic assays in enzymology. PMID- 15117196 TI - Ultraflat carbon film electrodes prepared by electron beam evaporation. AB - A facile method for the preparation of thin-film carbon electrodes by electron beam evaporation onto highly doped silicon is presented. The physical and electrochemical properties of these films both before and after postdeposition pyrolysis are investigated. Raman spectroscopy establishes the amorphous structure of the nonpyrolyzed carbon films and confirms the formation of graphitic carbon after pyrolysis at 1000 degrees C. Scanning force microscopy reveals the root-mean-square roughness of nonpyrolyzed films to be approximately 1 A, while pyrolyzed films exhibit an increased roughness of approximately 4 A. The electrochemical behavior of the electrodes resembles glassy carbon, with measured heterogeneous electron-transfer rate constants among the highest measured for thin carbon films. These carbon film electrodes will potentially find applications in such fields as molecular electronics and scanning probe microscopy of adsorbed species. PMID- 15117197 TI - Electrochemical performance of diamond thin-film electrodes from different commercial sources. AB - The electrochemical properties of two commercial (Condias, Sumitomo) boron-doped diamond thin-film electrodes were compared with those of two types of boron-doped diamond thin film deposited in our laboratory (microcrystalline, nanocrystalline). Scanning electron microscopy and Raman spectroscopy were used to characterize the electrode morphology and microstructure, respectively. Cyclic voltammetry was used to study the electrochemical response, with five different redox systems serving as probes (Fe(CN)(6)(3)(-)(/4)(-), Ru(NH(3))(6)(3+/)(2+), IrCl(6)(2)(-)(/3)(-), 4-methylcatechol, Fe(3+/2+)). The response for the different systems was quite reproducibile from electrode type to type and from film to film for electrodes of the same type. For all five redox systems, the forward reaction peak current varied linearly with the scan rate(1/2) (nu), indicative of electrode reaction kinetics controlled by mass transport (semi infinite linear diffusion) of the reactant. Apparent heterogeneous electron transfer rate constants, k degrees (app), for all five redox systems were determined from deltaE(p)-nu experimental data, according to the method described by Nicholson (Nicholson, R. S. Anal. Chem. 1965, 37, 1351.). The rate constants were also verified through digital simulation (DigiSim 3.03) of the voltammetric i-E curves at different scan rates. Good fits between the experimental and simulated voltammograms were found for scan rates up to 50 V/s. k degrees (app) values of 0.05-0.5 cm/s were observed for Fe(CN)(6)(3)(-)(/4)(-), Ru(NH(3))(6)(3+/2+), and IrCl(6)(2)(-)(/3)(-) without any extensive electrode pretreatment (e.g., polishing). Lower k degrees (app) values of 10(-)(4)-10(-)(6) cm/s were found for 4-methylcatechol and Fe(3+/2+). The voltammetric responses for Fe(CN)(6)(3)(-)(/4)(-) and Ru(NH(3))(6)(3+/2+) were also examined at all four electrode types at two different solution pH (1.90, 7.35). Since the hydrogen terminated diamond surfaces contain few, if any, ionizable carbon-oxygen functionalities (e.g., carboxylic acid, pK(a) approximately 4.5), the deltaE(p), i(p)(ox), and i(p)(red) values for the two systems were, for the most part, unaffected by the solution pH. This is in contrast to the typical behavior of oxygenated, sp(2) carbon electrodes, such as glassy carbon. PMID- 15117198 TI - Durable microfabricated high-speed humidity sensors. AB - We describe a durable microfabricated humidity sensor made of interdigitated rhodium electrodes on a silicon substrate covered with a sensing film of Nafion perfluorosulfonate ionomer. Rhodium electrodes are much less prone to oxidative degradation compared to previously described gold electrode-based sensors. Even with dc excitation, Rh electrode sensors exhibit excellent long-term response stability. It has been found that low-amplitude (+/-1 V) square wave excitation can prolong the usability of gold electrode-based sensors to at least several months; however, this mode of interrogation cannot provide subsecond response times. Rhodium deposition on the microsensors is much more difficult than that of gold. We were able to attain crack-free Rh deposits by adaptation of pulsed electroplating techniques. At excitation voltages of >2 V dc, the Rh sensors respond to moisture with 10 <--> 90% rise and fall times of 30-50 ms. These are the fastest microfabricated water vapor sensors reported to date. We demonstrate applications as a breath monitor. Such sensors should also be of utility in atmospheric eddy measurements. Short-term repeatability is better than 0.6% RSD (n = 7). PMID- 15117199 TI - Quantitative mass spectrometric determination of methylphenidate concentration in urine using an electrospray ionization source integrated with a polymer microchip. AB - We have demonstrated the use of a simple microfabricated electrospray ionization source for coupling microfluidic chips to mass spectrometry (MS). A polymer-based microchip, coupled to a triple quadrupole mass spectrometer, has been employed for direct infusion quantitative bioanalysis of methylphenidate (Ritalin) extracted from human urine samples. The approach used a microfabricated polymer electrospray emitter to couple a microfluidic channel to a stable electrospray ionization source. The microchip was fabricated from cycloolefin plastic plate by hot embossing and thermal bonding. This microfluidic chip contained two independent microfluidic channels, integrated with two corresponding electrospray emitters and an internal gold electrode. Liquid-liquid extraction was used to prepare urine samples, spiked with methylphenidate. A trideuterated analogue of methylphenidate (methylphenidate-d(3)) was used as the internal standard for the analysis. The system showed good electrospray stability and reproducibility with different spray tips. Four different electrospray tips were used to analyze the same sample, and the results showed very small variation with a relative standard deviation of 1.4%. A standard curve prepared for methylphenidate in urine (R(2) = 0.999) was linear over the range of 0.4-800 ng/mL. The precision of the quality control samples for three different concentrations ranged from 19.1% at 20 ng/mL, 3.2% at 200 ng/mL, to 3.5% at 667 ng/mL while the accuracy was 96.3% at 20 ng/mL, 101.2% at 200 ng/mL, and 101.6% at 667 ng/mL. No system carryover was detected even when the same device was used for sequential analysis. These results suggest the potential of this microdevice for MS-based quantitative analysis in drug discovery and development. PMID- 15117200 TI - Calibration-free estimates of batch process yields and detection of process upsets using in situ spectroscopic measurements and nonisothermal kinetic models: 4-(dimethylamino)pyridine- catalyzed esterification of butanol. AB - In this paper, we report the use of an NIR fiber-optic spectrometer with a high speed diode array for calibration-free monitoring and modeling of the reaction of acetic anhydride with butanol using the catalyst 4-(dimethylamino)pyridine in a microscale batch reactor. Acquisition of spectra at 5 ms/scan gave information relevant for modeling these fast batch processes with a single multibatch kinetic model. Nonlinear fitting of a first-principles model directly to the reaction spectra gave calibration-free estimates of time-dependent concentration profiles and pure component spectra. The amount of catalyst was varied between different batches to permit accurate estimation of its effect in the multiway model. A wide range of different models with increasing complexity could be fit to each batch individually with low residuals and apparent low lack of fit. However, only one model properly estimated the concentration profiles when all five batches were fitted simultaneously in a multiway kinetic model. Inclusion of on-line temperature measurements and use of an Arrhenius model for the estimated rate constant gave significantly improved model fits compared to an isothermal kinetic model. Augmentation of prerun batches with data from an additional batch permitted model-based forecasts of reaction trajectories, reaction yield, reaction end points, and process upsets. One batch with added water to simulate a process upset was easily detected by the calibration free process model. PMID- 15117201 TI - Pure component selectivity analysis of multivariate calibration models from near infrared spectra. AB - A novel procedure is proposed as a method to characterize the chemical basis of selectivity for multivariate calibration models. This procedure involves submitting pure component spectra of both the target analyte and suspected interferences to the calibration model in question. The resulting model output is analyzed and interpreted in terms of the relative contribution of each component to the predicted analyte concentration. The utility of this method is illustrated by an analysis of calibration models for glucose, sucrose, and maltose. Near infrared spectra are collected over the 5000-4000-cm(-)(1) spectral range for a set of ternary mixtures of these sugars. Partial least-squares (PLS) calibration models are generated for each component, and these models provide selective responses for the targeted analytes with standard errors of prediction ranging from 0.2 to 0.7 mM over the concentration range of 0.5-50 mM. The concept of the proposed pure component selectivity analysis is illustrated with these models. Results indicate that the net analyte signal is solely responsible for the selectivity of each individual model. Despite strong spectral overlap for these simple carbohydrates, calibration models based on the PLS algorithm provide sufficient selectivity to distinguish these commonly used sugars. The proposed procedure demonstrates conclusively that no component of the sucrose or maltose spectrum contributes to the selective measurement of glucose. Analogous conclusions are possible for the sucrose and maltose calibration models. PMID- 15117202 TI - Detection of cytochrome C in a single cell using an optical nanobiosensor. AB - In this work, the intracellular measurement of cytochrome c using an optical nanobiosensor is demonstrated. The nanobiosensor is a unique fiberoptics-based tool which allows the minimally invasive analysis of intracellular components. Cytochrome c is a very important protein to the process which produces cellular energy. In addition, cytochrome c is well-known as the protein involved in apoptosis, or programmed cell death. delta-Aminolevulinic acid (5-ALA) was used to induce apoptosis in MCF-7 human breast carcinoma cells. 5-ALA, a photodynamic therapy (PDT) drug in cells was activated by a HeNe laser beam. After the PDT photoactivation, the release of cytochrome c from the mitochondria to the cytoplasm in a MCF-7 cell was monitored by the optical nanobiosensor inserted inside the single cell and followed by an enzyme-linked immunosorbent assay (ELISA) outside the cell. The combination of the nanobiosensor with the ELISA immunoassay improved the detection sensitivity of the nanobiosensor due to enzymatic amplification. Our results lead to the investigation of an apoptotic pathway at the single cell level. PMID- 15117203 TI - Transfer of multivariate calibrations between four near-infrared spectrometers using orthogonal signal correction. AB - The transfer of partial least squares (PLS) calibration models among four near infrared spectrometers was investigated for the quantitative analysis of thermoset resin polymers. A comparative study of second derivatives, multiplicative scatter correction, finite impulse response filtering, slope and bias correction, model updating (MU), and orthogonal signal correction (OSC) was conducted to determine which processing methods achieved model transferability. It is shown that OSC and MU were superior to the other calibration transfer methods, leading to very robust PLS models with enhanced predictive ability. It is also shown that the transfer results obtained with OSC were not significantly different from those obtained with model updating. PMID- 15117204 TI - Evaluation of the uniformity of analytical-size chromatography columns prepared by the downward packing of particulate slurries. AB - The axial heterogeneity of downward slurry-packed chromatography columns was evaluated. A series of columns were prepared that varied in total length from 5 to 30 cm. Each column was packed in stainless steel tubing that was sectioned to allow the column to be divided into 5-cm sections after packing. Each 5-cm section of these columns was then tested for chromatographic performance. In total, 22 such column sections were tested. The results of the study show that the consolidation of the packed bed is a very complex problem, yet systematic variations of the column performance were observed, depending on the location of the section tested from within the entire column. For example, when columns longer than 20 cm were packed, the most homogeneous and best performing section of the column was the lower midsection. Whereas for columns shorter than 20 cm in length, the best sectioned region of the bed was the lower and outlet region. In all cases, the most poorly packed region of the bed was the column inlet section, irrespective of the bed length. The phenomenon associated with this axial heterogeneity undoubtedly results from a complex interplay between wall friction, particle momentum, and the pressure pulsations resulting from the packing process. PMID- 15117205 TI - Systematization of the mass spectra for speciation of inorganic salts with static secondary ion mass spectrometry. AB - The analytical use of mass spectra from static secondary ion mass spectrometry for the molecular identification of inorganic analytes in real life surface layers and microobjects requires an empirical insight in the signals to be expected from a given compound. A comprehensive database comprising over 50 salts has been assembled to complement prior data on oxides. The present study allows the systematic trends in the relationship between the detected signals and molecular composition of the analyte to be delineated. The mass spectra provide diagnostic information by means of atomic ions, structural fragments, molecular ions, and adduct ions of the analyte neutrals. The prediction of mass spectra from a given analyte must account for the charge state of the ions in the salt, the formation of oxide-type neutrals from oxy salts, and the occurrence of oxidation-reduction processes. PMID- 15117206 TI - Fingerprinting metal-containing biomolecules after reductive displacement of iron by gallium and subsequent column-switched LC-ICPMS analysis applied on siderophores. AB - Column-switching liquid chromatography followed by low-resolution ICPMS was evaluated as a tool for speciation analysis of metal-containing biomolecules. The strategy was applied on siderophores, strong iron chelators of low molecular weight (M(w) < 1500). Prior to the LC-ICPMS analysis, reductive displacement of iron by gallium was performed using ascorbate as the reducing agent to increase the sensitivity. Different experimental conditions during the exchange reaction were tested using ferrichrysin and ferrichrome for evaluation. A reaction time of 30 min and a pH of 3.9 gave an exchange yield of 27 and 83% for ferrichrysin and ferrichrome, respectively. A gradient elution profile was also developed to separate gallium-chelated siderophores on a PGC column. Detection limits for standard solutions of ferrichrysin and ferrichrome in the low-nanomolar range were obtained by monitoring the gallium-69 isotope. The combined use of LC-ICPMS and LC-ESI-MS/MS was also evaluated as a tool to identify unknown metal complexes, here siderophores, in field soil solution samples. PMID- 15117207 TI - Hyphenation of capillary HPLC to microcoil (1)H NMR spectroscopy for the determination of tocopherol homologues. AB - Highly selective reversed phases (C(30) phases) are self-packed in 250 microm inner diameter fused-silica capillaries and employed for capillary HPLC separation of shape-constrained natural compounds (tocopherol homologues, vitamin E). Miniaturized hyphenated systems such as capillary HPLC-ESI-MS (positive ionization mode) and, with special emphasis, continuous-flow capillary HPLC- NMR are used for structural determination of the separated compounds. Despite the small amount of sample available (1.33 microg of each tocopherol), the authors have been able to monitor the capillary HPLC separation under continuous-flow (1)H NMR conditions, thus allowing an immediate peak identification. Further structural assignment was carried out in the stopped-flow NMR mode as shown, for example, by a 2D (1)H,(1)H COSY NMR spectrum of alpha-tocopherol. We demonstrate in this paper the considerable potential of hyphenated capillary separations coupled to MS and NMR for the investigation of restricted amounts of sample. PMID- 15117208 TI - Design, fabrication, and evaluation of microfabricated columns for gas chromatography. AB - The design, fabrication, and performance of gas chromatography columns etched in silicon substrates are described. Deep reactive-ion etching formed the 3-m-long, 150-microm-wide, 240-microm-deep rectangular cross section channels. A glass cover plate was anodically bonded to the remaining surface of the substrate forming the gastight channel. For some of the columns, the silicon channels were oxidized before the channels were sealed with the glass plates. Fused-silica capillary connecting tubes were sealed into ports on the edge of the 3.2-cm x 3.2 cm substrate chips. Dynamic coating was used to deposit a film of nonpolar dimethyl polysiloxane or moderately polar trifluoropropylmethyl polysiloxane stationary phase. The columns were evaluated in a conventional benchtop GC instrument with split injection and flame ionization detection. Column efficiency was evaluated by the use of plots of height equivalent to a theoretical plate versus average carrier gas velocity using both hydrogen and air as carrier gases. The number of theoretical plates measured at the average carrier gas velocity giving the minimum plate height ranged from 4600 to 8200 plates for the dimethyl polysiloxane columns and from 3500 to 5500 plates for the trifluoropropylmethyl polysiloxane columns. Minimum plate height was significantly smaller with air as carrier gas. For the nonpolar phase, the nonoxidized surface gave approximately 1500 plates more than the oxidized surface for both carrier gases. For the polar phase, the oxidized surface gave approximately 200 plates more than the nonoxidized surface. Isothermal chromatograms of a 20-component multifunctional mixture and temperature-programmed chromatograms of a normal alkane mixture are presented. PMID- 15117209 TI - HPLC and MALDI-TOF MS analysis of highly branched polystyrene: resolution enhancement by branching. AB - Temperature gradient interaction chromatography (TGIC) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) were applied for the characterization of highly branched polystyrenes (PS) prepared by linking living polystyryl anions using 4-chlorodimethylsilylstyrene. Reversed phase (RP)-TGIC showed an unexpectedly high resolution according to the number of branches despite significant overlap of the molecular weight as confirmed by MALDI-TOF MS. The enhancement of the resolution is ascribed to the contribution of the nonpolar groups in the branched PS: the dimethylsilyl groups in the branching unit as well as the sec-butyl initiator groups. As the number of branches increases, the number of nonpolar groups increases, which in turn increases the RP-TGIC retention synergistically with increasing molecular weight. In contrast, a poorer resolution was found in normal-phase-TGIC, in which the nonpolar groups reduce the retention. The resolution in RP-TGIC appears superior to that of liquid chromatography at the chromatographic critical condition (LCCC) of PS. It is seemingly due to the synergistic contribution of the incremental PS molecular weight to the functionality in the branched PS in RP-TGIC while only the functionality contributes to the separation in LCCC. This type of resolution enhancement could be utilized efficiently for the analysis of highly branched polymers such as dendrimers or hyperbranched polymers. PMID- 15117210 TI - Simultaneous quantitative determination of benzene, toluene, and xylenes in water using mid-infrared evanescent field spectroscopy. AB - Attenuated total reflection mid-infrared spectroscopy is applied for simultaneous detection and quantification of the environmentally relevant analytes benzene, toluene, and the three xylene isomers. The analytes are enriched into a thin polymer membrane coated onto the surface of an internal reflection waveguide, which is exposed to the aqueous sample. Direct detection of analytes permeating into the polymer coating is performed by utilizing evanescent field spectroscopy in the fingerprint range (>10 microm) of the mid-infrared (MIR) spectrum (3-20 microm) without additional sample preparation. All investigated compounds are characterized by well-separated absorption features in the evaluated wavelength regime. Hence, data evaluation was performed by integration of the respective absorption peaks. Limits of detection lower than 20 ppb (v/v) for all xylene isomers, 45 ppb (v/v) for benzene, and 80 ppb (v/v) for toluene have been achieved. The straightforward experimental setup and the achieved detection limits for these environmentally relevant volatile organic compounds in the low ppb concentration range reveal a substantial potential of MIR evanescent field sensing devices for on-line in situ environmental analysis. PMID- 15117211 TI - Evaluation of 2-methacryloyloxyethyl phosphorylcholine polymeric nanoparticle for immunoassay of C-reactive protein detection. AB - To prepare novel 2-methacryloyloxyethyl phosphorylcholine (MPC)-polymeric nanoparticle (MPC-PNP), water-soluble amphiphilic phospholipid polymer, poly [MPC co-n-butyl methacrylate (BMA)-co-p-nitrophenyloxycarbonyl poly(ethylene glycol) methacrylate (MEONP) (PMBN)], which has active ester groups for bioconjugation on the side chains, was synthesized. MPC-PNP was prepared by a solvent evaporation technique where the poly(l-lactic acid) was used as core and PMBN was applied as an emulsifier and a surface modifier under systematical design of well-arranged phospholipids polar groups in its surface. Characteristics for MPC-PNP were thoroughly investigated with dynamic light scattering, electrophoresis light scattering, X-ray photoelectron spectroscopy, and field emission scanning electron microscopy measurements. Through a protein adsorption test, the phosphorylcholine group on the surface of MPC-PNPs, which had their active ester groups substituted by glycine, were shown to suppress the nonspecific adsorption of bovine serum albumin. These particles were used for C-reactive protein (CRP) detection, where anti-CRP monoclonal antibodies were immobilized on the MPC-PNP using the active ester group, while the remaining active ester groups were thoroughly reacted with glycine. The detection limit about serum-free CRP in the calibration curve was shown to extend from 0.01 to 10 mg/dL when anti-CRP antibody immobilized MPC-PNP was used for serum-free CRP detection. This compares favorably with measurement using polystyrene nanoparticles that were shown to detect from 0.1 to 10 mg/dL by an immunoagglutination technique. Also, for the detection of CRP in serum, MPC-PNP was shown to give the same calibration curve explained by the efficient suppression of nonspecific binding. Furthermore, denaturation of immobilizing anti-CRP antibody on the MPC-PNP hardly occurred despite increasing the temperature. It is concluded that MPC-PNP is unique due to the design of its interfacial properties, also it will perform well in a diagnostic immunoassay because of its optimized material properties. PMID- 15117212 TI - Performance optimization of spectroscopic process analyzers. AB - To increase the power and the robustness of spectroscopic process analyzers, methods are needed that suppress the spectral variation that is not related to the property of interest in the process stream. An approach for the selection of a suitable method is presented. The approach uses the net analyte signal (NAS) to analyze the situation and to select methods to suppress the nonrelevant spectral variation. The empirically determined signal-to-noise of the NAS is used as a figure of merit. The advantages of the approach are (i). that the error of the reference method does not affect method selection and (ii). that only a few spectral measurements are needed. A diagnostic plot is proposed that guides the user in the evaluation of the particular suppression method. As an example, NIR spectroscopic monitoring of a mol-sieve separation process is used. PMID- 15117213 TI - A theoretical approach to some analytical properties of heterogeneous enzymatic assays. AB - Heterogeneous enzymatic assays (HEA), where an enzyme in solution acts upon an immobilized substrate, are been increasingly used. Given their high throughput and versatility they hold great potential for developing massive enzyme inhibitor screening. However, current HEA lack, in general, rigorous quantitative use. This is in part due to technical problems as a multiplicity of suboptimal substrate populations achieved with traditional immobilization techniques but, more importantly, is due to a poor understanding of the particular kinetic behavior of these systems. This paper addresses the kinetic features of HEA that arise from the very low amount of solid-phase substrate and the resulting inalterability of the free enzyme concentration during the assay, which classify HEA as enzyme quasi-saturable systems (EQSS). We assessed the optimal enzyme concentration working range and time of reaction. We also considered certain attributes of HEA for evaluating isosteric inhibitors. These studies were done on the basis of a simplified model for the kinetics of EQSS and a formal splitting of the functional factor of the analytical sensitivity of an enzymatic assay into [E(o)]/K(m)-dependent and temporal components. PMID- 15117214 TI - Direct determination of the peptide content in microspheres by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - A quantitative determination of peptides incorporated into poly(d,l-lactide-co glycolide) microspheres by matrix-assisted laser desorption/ionization time-of flight mass spectrometry (MALDI-TOF MS) was accomplished in a single step without pretreatment for extracting the peptide from the microsphere. The conventional extraction methods often underestimate the actual amount of peptide because of incomplete extraction from the microspheres or loss during the procedures. In this study, the microspheres dissolved in acetonitrile containing 0.1% trifluoroacetic acid were mixed with matrix solution containing the internal standard, and the peptide content was directly determined by MALDI-TOF MS. The drug content values determined by MALDI-TOF MS in both the leuprolide- and salmon calcitonin-incorporated microspheres were closer to the theoretical contents than those determined by the conventional extraction method. This method using MALDI TOF MS could be a good alternative to time-consuming and less-accurate conventional methods. PMID- 15117215 TI - On the absence of evidence that the Vinland map is medieval. AB - The article, Evidence That the Vinland Map Is Medieval (Anal. Chem. 2003, 75, 6745-6747), is constructed with conjecture and with false logic. It imagines that the ink of the Vinland Map is an iron-gall ink where conclusive evidence shows the ink to be not an iron-gall ink but a carbon-based ink. It asserts that the ink was homogeneous where conclusive evidence shows that it is not. Twenty-nine years ago, the Vinland Map was shown incontrovertibly to be modern. That conclusion stands today. PMID- 15117219 TI - States' initial response to the President's New Freedom Initiative: slowest rates of deinstitutionalization in 30 years. PMID- 15117222 TI - Thoughts after the fatal beating of Ricky Whistnant. PMID- 15117223 TI - Living outside the system: the ups and downs of getting on with our lives. PMID- 15117224 TI - Community inclusion of individuals with behavioral challenges: who supports the careproviders? PMID- 15117225 TI - Ethical analysis of the concept of disability. AB - The concept of disability from an ethical viewpoint was examined. Whether disability results from the way society is built and the way ideas and judgments considering disability are established in a communal interaction was discussed (i.e., is disability socially created and constructed?). Although views on disability emphasizing its social nature are basically sound, they are also insufficient because they fail to consider the normative dimension attached to the concept of disability. The core of the concept of disability is ethical, which is why a moral philosophical examination of the concept is needed. Three possible ethical theories of disability (universal, objective, subjectivist, and communitarian) are briefly presented and discussed and a preliminary outline of a basis for a sound ethical model of disability presented. PMID- 15117226 TI - Intellectual and physical disabilities in prehistory and early civilization. AB - This paper is focused on three basic questions: The first concerns when specific disabilities first appeared during human evolution. The second question has to do with causes of disabilities. The third question concerns social responses to people with disabilities. Discussions on each of the issues are presented. PMID- 15117227 TI - Perspectives of physicians, families, and case managers concerning access to health care by individuals with developmental disabilities. AB - This study of the status of medical care for Kansans with developmental disabilities consists of reports from physicians, service providers, and family members. Overall, these three groups indicated satisfaction with medical care across the four criteria of availability, accessibility, appropriateness, and affordability. The bases for these results are outlined, and suggestions for improving satisfaction with health care are presented. PMID- 15117228 TI - Person-centered services and organizational context: taking stock of working conditions and their impact. AB - Although the concept of person-centered services has gained much popularity, there are significant discrepancies in the extent to which service agencies are actually implementing it. Eight organizations, each of which was successfully providing person-centered services to at least some individuals, were examined. Some agencies were implementing the approach with significantly higher proportions of individuals receiving their services than were others. Interviews and participant observations were used to examine how the various organizational contexts contributed to these differences. Some organizations were found to be well-suited to the approach, whereas others posed significant limitations to the number of individuals who benefited or the rate at which they were likely to benefit from this approach. PMID- 15117230 TI - Osteoblasts subjected to spaceflight and simulated space shuttle launch conditions. AB - To understand further the effects of spaceflight on osteoblast-enriched cultures, normal chicken calvarial osteoblasts were flown aboard shuttle flight STS-77, and the total number of attached cells was determined. Spaceflight and control cultures were chemically fixed 3 h and 3 d after launch. These fixed cultures were processed for scanning electron microscopy (SEM). The SEM analysis showed that with just 3 d of exposure to spaceflight, coverslip cultures contained 300 +/- 100 cells/mm2, whereas 1G control samples contained a confluent monolayer of cells (2400 +/- 200 cells/mm2). Although the cultures flown in space experienced a drastic decline in cell number in just 3 d, without further experimentation it was impossible to determine whether the decline was a result of microgravity, the harsh launch environment, or some combination of these factors. Therefore, this research attempted to address the effect of launch by subjecting osteoblasts to conditions simulating shuttle launch accelerations, noise, and vibrations. No differences, compared with controls, were seen in the number of total or viable cells after exposure to these various launch conditions. Taken together, these data indicate that the magnitude of gravitational loading (3G maximum) and vibration (7.83G rms maximum) resulting from launch does not adversely affect osteoblasts in terms of total or viable cell number immediately, but launch conditions, or the microgravity environment itself, may start a cascade of events that over several d contributes to cell loss. PMID- 15117231 TI - Establishment of hepatic stem-like cell lines from normal adult porcine liver in a poly-D-lysine-coated dish with NAIR-1 medium. AB - The existence, origin, and bipotency of the hepatic stem cell (HeSC) have been investigated. However, the isolation and culture of HeSCs from adult liver tissue is not yet well established, and the mechanism by which HeSCs differentiate into mature cells remains unclear. On the other hand, the development of HeSC isolating and -culturing methods and the in vitro clonal analysis of their mechanism of differentiation are required to enable clinical applications of regenerative medicine in the liver. For the purpose of providing HeSCs for these studies, we attempted to establish an HeSC line from a normal adult porcine liver using a unique culture system, a poly-D-lysine-coated culture dish with NAIR-1 medium (the PDL-NAIR-1 culture system). Moreover, we examined the differentiating capacity of HeSCs in vitro. We demonstrated that it was possible in the culture system that immature epithelial cells capable of proliferating grew selectively into aggregates and that two hepatic stem-like cell lines, PHeSC-A1 and PHeSC-A2, were established. The results from our data suggest that these hepatic stem-like cell lines were capable of self-renewing and differentiating into hepatocytes or biliary epithelial cells and show that the PDL-NAIR-1 culture system offers the immense advantage of isolating and culturing HeSCs from a normal adult liver. Furthermore, because of the ability to use a clonal analysis in vitro, these cell lines are useful for the investigation of various mechanisms in which HeSCs seem to participate and their application in the study of regenerative medicine in the liver. PMID- 15117232 TI - Differential expression of mammalian or viral promoter-driven gene in adherent versus suspension cells. AB - Although expression vectors using viral and mammalian promoters constitutively express genes of interest in adherent cells, few studies have examined whether the function of these vectors in suspended cells, such as in over-agar or soft agar assay (an in vitro cell transformation assay), is as robust as when they are in adherent cells. The selection of appropriate expression vector to optimally express genes in suspended cells would be useful in determining whether these genes play a critical role in maintaining colony formation or cell transformation. To compare promoter-driven expression vector function in adherent versus suspension cells, we performed transient transfection assays using viral (simian virus 40 [SV40] and cytomegalovirus [CMV]) and mammalian (beta-actin) promoters fused to luciferase or beta-galactosidase reporter gene. Over-agar assay was used to suspend cells on top of agar, which allowed cell retrieval and analysis. We found that beta-actin and SV40 promoters exhibited suppressed gene expression of 70 and 56%, respectively, in cells suspended on agar compared with those attached on plates. The suppressed response by the exogenous beta-actin promoter in suspension was consistent with the response of the endogenous beta actin promoter activity because the steady-state level of beta-actin messenger ribonucleic acid in suspended cells was significantly reduced by 50% relative to that expressed in attached cells. In contrast to SV40 promoter, CMV promoter activity was not decreased in cells suspended in over-agar when compared with adherent cells. These studies show that regardless of mammalian or viral vectors, one cannot assume that all expression vectors behave similarly in both suspension and adherent state. PMID- 15117233 TI - A continuous cell line from the cupreous chafer, Anomala cuprea Hope (Insecta, Coleoptera, Scarabaeidae). AB - A continuous cell line was obtained from the culture of embryonic cells of the cupreous chafer, Anomala cuprea Hope. The cells showed substrate-dependent growth and formed loose networks. Population doubling time was about 4.5 d. The mode of chromosome number was about 32 (4n). The cell line was designated FRI-AnCu-35. PMID- 15117235 TI - Cholelitiasis and gastroesophageal reflux. PMID- 15117236 TI - Non-complicated cholelithiasis associated with GERD. Results of combined laparoscopic surgery in low risk patients. AB - BACKGROUND: the aim of this study was to evaluate the efficacy of combined laparoscopic surgery for non-complicated cholelithiasis and gastroesophageal reflux disease (GERD) in patients with low surgical risk. METHODS: a total of 680 cholecystectomies performed by means of laparoscopic surgery were retrospectively studied from February 1991 to February 2002. A total of 442 patients that fulfilled the inclusion criteria were divided into two groups: group A: non complicated cholelithiasis (cholecystectomy alone), consisting of a total of 362 patients, and group B: non-complicated cholelithiasis and GERD (cholecystectomy and Toupet's fundoplication in all cases) in 80 patients. Demographic and clinical data, intraoperatory incidences, and post-surgical complications were prospectively collected and compared for all patients. The results of reflux surgery (group B) were evaluated at 6 months by means of 24-hour pH-metry. RESULTS: in spite of the fact that the group undergoing combined surgery consisted of patients with greater weight and older age (p < 0.05), no significant differences were found in the number of intraoperative incidences and post-surgical complications between both groups (NS). Significant differences were only found in the duration of surgery: 48 +/- 25 min (10-150) in group A compared to 112 +/- 23 min (80-180) in group B (p<0.001), and in the return to normal daily activities (5.8 +/- 0.9 days vs 6.5 +/- 1 days in group B) (p< 0.001). In the latter group a normalization of 24-hour pH-metry values and an absence of symptoms associated with reflux were observed in all cases. CONCLUSIONS: in patients younger than 75 years with low surgical risk and non complicated cholelithiasis and GERD, both illnesses can be resolved during the same surgical procedure by laparoscopy with no increased risk or postoperative complications. PMID- 15117237 TI - Tissue CA-19.9 content in colorectal adenomas and its value in the assessment of dysplasia. AB - BACKGROUND: occasionally, the risk of malignant transformation may be difficult to establish in adenomatous polyps due to the fact that they contain areas with variable grades of dysplasia. A measurement of tissue tumor markers may be useful to recognize these adenomas. OBJECTIVES: the aims of this study were: to established firstly the relationship between carbohydrate antigen 19.9 (CA-19.9) content in the colorectal mucosa and the characteristics of polyps, and secondly, the diagnostic value of the formers measurement. PATIENTS AND METHODS: tissue CA 19.9 concentration was measured in 155 colorectal samples obtained from 145 patients (21 normal mucosa; 113 adenomatous polyps; 21 adenocarcinoma). Cytosol CA-19.9 content was determined by enzyme-linked immunoadsorbant assay, and the measurement of this protein was achieved by quantitative assay. Tissue samples were also processed for histological examination. RESULTS: we demonstrated that CA-19.9 levels in adenomatous polyps and adenocarcinomas were significantly higher than in the normal mucosa. These levels varied significantly according to polyp size, histological type, and grade of dysplasia. CA-19.9 contents were higher in polyps with a high risk of malignant transformation than in those with a low risk of severe dysplasia. The cut-off value 214 U/mg of protein properly differentiated both types of risk. The area under the receiver operating characteristic (ROC) curves showed that cytosol CA-19.9 levels allow classifying polyps according to their histological features. CONCLUSIONS: we concluded that the measurement of CA-19.9 content in adenomatous polyps may be useful to classify these tumors and confirm the feasibility to separate adenomas into two groups: low and high risk of malignant change. PMID- 15117238 TI - Topical glyceryl trinitrate in the treatment of anal fissure. AB - AIM: the aim of this study was to assess the effectiveness of topical glyceryl trinitrate (GTN) in the treatment of anal fissure. PATIENTS: eighty consecutive patients were randomized to receive 0.2% GTN gel twice a day during a minimum of 6 weeks. Of these 80 patients 34 were males and 46 female, with ages between 20 and 78 years. All patients underwent a work-up that included gender, age, symptom duration, site, previous treatment attempts, pain score, associated diseases, and maximum anal resting pressure. These patients were followed at regular intervals of 4, 8, and 12 weeks to assess symptom outcome, rate of healing, adverse effects, and recurrence rate. RESULTS: pain scores were significantly reduced during the treatment period in 65% of cases. After 4 weeks, 55% of patients had healed, and 78% after 9 weeks. In 18 patients (22%), anal fissures did not heal even with 6 additional weeks of treatment, and 12 of them (15%) underwent lateral sphincterotomy. Sixty one percent of patients had flushing and 15% severe headaches. CONCLUSION: the results of this study have demonstrated the significant benefit of topical GTN when administered to patients suffering from anal fissures. PMID- 15117239 TI - Usefulness of jejunal biopsy in the study of intestinal malabsorption in the elderly. AB - BACKGROUND: small bowel structure and function are not different between elderly people and young people. Thus, in principle it is advisable to perform diagnostic investigation of elderly patients as well as younger patients when they present with symptoms suggestive of intestinal malabsorption. A key test for the etiologic diagnosis of intestinal malabsorption, jejunal biopsy, has not been specifically examined to assess its usefulness and risk of complications in this advanced age patients. AIM: to establish the usefulness of jejunal biopsy with the Watson's capsule in the elderly patients with suspected intestinal malabsorption. PATIENTS: patients older than 65 years referred to our Unit for performance of a jejunal biopsy from 1996 to 2001 for suspicion of intestinal malabsorption. RESULTS: forty-seven patients were included. Appropriate biopsy sample was obtained in 45 cases, although in 3 patients a second try was required. Histologic findings: partial villous atrophy in 10 cases (22.2%), complete villous atrophy in 5 cases (11.1%), intraepithelial lymphocytosis in 5 cases (11.1%), and single cases of intestinal lymphangiectasia, amyloidosis, unspecific jejunitis, and Whipple's disease. Histology was normal in 19 cases (42%). Definitive diagnosis was celiac disease in 14 patients, bacterial overgrowth in 3, jejunitis in 3, Whipple's disease in 1, lymphangiectasia in 1, atrophic gastritis in 3, amyloidosis in 1, and ischemic colitis in 1. Jejunal biopsy achieved an etiologic diagnosis in 20 patients. There were no cases of perforations or bleeding. CONCLUSION: jejunal biopsy is a useful and safe test for the etiologic diagnosis of intestinal malabsorption in elderly patients. PMID- 15117245 TI - Serendipitous meanderings and adventures with molecular beams. AB - This is the story of a native-born American who came as a postdoc to the country of his parents, Germany. There, by good fortune, he could participate in the revival and the rebuilding of the physical sciences following the ravishments of the Second World War, becoming at the age of 38, the director of a Max-Planck Institut in Gottingen. Working under nearly ideal conditions, he carried out basic research using molecular beams. Aided by many active, youthfully impulsive, yet perceptive and imaginative, students and experienced knowledgeable guest scientists from many countries, he enjoyed exciting adventures into unknown landscapes in the fields of molecular gas-phase interactions and solid-surface phenomena and, most recently, in the realms of quantum liquids and solids. PMID- 15117246 TI - Surface chemistry and tribology of MEMS. AB - The microscopic length scale and high surface-to-volume ratio, characteristic of microelectro-mechanical systems (MEMS), dictate that surface properties are of paramount importance. This review deals with the effects of surface chemical treatments on tribological properties (adhesion, friction, and wear) of MEMS devices. After a brief review of materials and processes that are utilized in MEMS technology, the relevant tribological and chemical issues are discussed. Various MEMS microinstruments are discussed, which are commonly employed to perform adhesion, friction, and wear measurements. The effects of different surface treatments on the reported tribological properties are discussed. PMID- 15117247 TI - Formation of novel rare-gas molecules in low-temperature matrices. AB - Progress in the study of a new class of chemically bound compounds of noble-gas atoms is reviewed. The focus is on rare-gas molecules of the form HNgY, where Ng is a noble-gas atom and Y is an electronegative group, prepared by photolysis of HY in the rare-gas matrix. Other related types of new molecules of noble-gas atoms are discussed as well. Topics discussed in this review include: (a) The nature of bonding and the energetic stability of the compounds. (b) The vibrational spectroscopy of the molecules, and its role in identification of the species. (c) The mechanism and dynamics of photochemical formation of HNgY in the matrix, and the pathways for thermal and infrared (IR)-induced decomposition. Specifically, attention is given to the issue of "direct" formation following photolysis of HY versus "delayed" formation involving H atom diffusion. (d) Molecules of the lighter rare gases Ar, Ne, and He, focusing on the experimentally prepared HArF and on theoretical predictions suggesting the existence of other molecules. (e) The most-recently discovered photochemically induced insertion compounds of Ng into hydrocarbons, such as HXeCCH. (f) Clusters of HNgY with other molecules. The possible existence of neat aggregates and crystals of HNgY. The reviewed state-of-the-art suggests this field is at an early stage of development with major open questions bearing on the surprising properties of the molecules and on the formation mechanisms. These are part of the challenge for the future. PMID- 15117248 TI - Single-molecule fluorescence spectroscopy and microscopy of biomolecular motors. AB - The methods of single-molecule fluorescence spectroscopy and microscopy have been recently utilized to explore the mechanism of action of several members of the kinesin and myosin biomolecular motor protein families. Whereas ensemble averaging is removed in single-molecule studies, heterogeneity in the behavior of individual motors can be directly observed, without synchronization. Observation of translocation by individual copies of motor proteins allows analysis of step size, rate, pausing, and other statistical properties of the process. Polarization microscopy as a function of nucleotide state has been particularly useful in revealing new and highly rotationally mobile forms of particular motors. These experiments complement X-ray and biochemical studies and provide a detailed view into the local dynamical behavior of motor proteins. PMID- 15117249 TI - Dynamics of single biomolecules in free solution. AB - Instrumental advances have allowed the continuous observation of single-molecule trajectories in free solution. Diffraction-limited spectral resolution at video frame rates is routinely achieved by using commercial, intensified, charge coupled device cameras, low-power continuous-wave lasers, and standard optical microscopes. Either the native fluorescence from large biomolecules or emission from conjugated fluorescence labels can be employed to follow multiple molecules over many seconds. Both molecular motion at the liquid/solid interface and in bulk solution can be recorded. The former reveals adsorption and desorption probabilities that are related to chromatographic retention processes and to the applicability of biocompatible materials. The latter allows the manipulation of particles and large biomolecules to facilitate separation and identification. PMID- 15117250 TI - Beyond Born-Oppenheimer: molecular dynamics through a conical intersection. AB - Nonadiabatic effects play an important role in many areas of physics and chemistry. The coupling between electrons and nuclei may, for example, lead to the formation of a conical intersection between potential energy surfaces, which provides an efficient pathway for radiationless decay between electronic states. At such intersections the Born-Oppenheimer approximation breaks down, and unexpected dynamical processes result, which can be observed spectroscopically. We review the basic theory required to understand and describe conical, and related, intersections. A simple model is presented, which can be used to classify the different types of intersections known. An example is also given using wavepacket dynamics simulations to demonstrate the prototypical features of how a molecular system passes through a conical intersection. PMID- 15117251 TI - Functional oxide nanobelts: materials, properties and potential applications in nanosystems and biotechnology. AB - Nanobelt is a quasi-one-dimensional structurally controlled nanomaterial that has well-defined chemical composition, crystallographic structure, and surfaces (e.g., growth direction, top/bottom surface, and side surfaces). This article reviews the nanobelt family of functional oxides, including ZnO, SnO2, In2O3, Ga2O3, CdO, and PbO2 and the relevant hierarchical and complex nanorods and nanowires that have been synthesized by a solid-vapor process. The nanobelts are single crystalline and dislocation free, and their surfaces are atomically flat. The oxides are semiconductors that have been used for fabrication of nanosize functional devices of key importance for nanosystems and biotechnology, such as field-effect transistors, gas sensors, nanoresonators, and nanocantilevers. The structurally controlled ZnO nanobelts that exhibit piezoelectric properties are also reviewed. By controlling growth kinetics, we show the success of growing nanobelt-based novel structures whose surfaces are dominated by the polarized + (0001) facets. Owing to the positive and negative ionic charges on the zinc- and oxygen-terminated +-(0001) surfaces, respectively, a spontaneous polarization is induced across the nanobelt thickness. As a result, helical nanostructures and nanorings are formed by rolling up single-crystal nanobelts; this phenomenon is a consequence of minimizing the total energy contributed by spontaneous polarization and elasticity. The polar surface-dominated ZnO nanobelts are likely to be an ideal system for understanding piezoelectricity and polarization-induced ferroelectricity at nano-scale and they could have applications as one dimensional nano-scale sensors, transducers, and resonators. PMID- 15117252 TI - Adsorption and reaction at electrochemical interfaces as probed by surface enhanced Raman spectroscopy. AB - Over the past three decades, surface-enhanced Raman spectroscopy (SERS) has gone through a tortuous pathway to develop into a powerful surface diagnostic technique for in situ investigation of surface adsorption and reactions on electrodes. This review presents the recent progress achieved mainly in our laboratory on the improvement of detection sensitivities as well as spectral, temporal, and spatial resolutions. Various surface roughening procedures for electrodes of different metals coupled with maximum use of a high-sensitivity confocal Raman microscope enable us to obtain good-quality SER spectra on the electrode surfaces made from net Pt, Ni, Co, Fe, Pd, Rh, Ru, and their alloys that were traditionally considered to be non-SERS active. A novel technique called potential-averaged SERS (PASERS) has been developed for the quantitative study of electrochemical sorption. Applications are exemplified on extensively studied areas such as coadsorption, electrocatalysis, corrosion, and fuel cells, and several advantages of in situ electrochemical SERS are demonstrated. Finally, further developments in this field are briefly discussed with emphasis on the emerging methodology. PMID- 15117253 TI - Molecular beam studies of gas-liquid interfaces. AB - Molecular beam scattering experiments provide a way to disentangle the elementary steps involved in energy transfer and chemical reactions between gases and liquids. After surveying the history and recent progress in this field, we review studies of the kinematics of gas-liquid collisions and proton exchange of HCl, DCl, and HBr with supercooled sulfuric acid and liquid glycerol. These experiments help to clarify the role of the surface region in controlling trapping and interfacial- and bulk-phase reactions. PMID- 15117254 TI - Charge transport at conjugated polymer-inorganic semiconductor and conjugated polymer-metal interfaces. AB - Charge transport at conjugated polymer interfaces with metals and inorganic semiconductors is reviewed. Experiments on the equilibrium properties and DC current-voltage behavior of four specific classes of interfaces-metal-doped conjugated polymer, inorganic semiconductor-doped conjugated polymer, metal intrinsic conjugated polymer, and metal-intrinsic conjugated polymer/electrolyte are discussed. To facilitate this discussion, classic models of equilibration at ideal interfaces between electronic conductors and free-electron transport are introduced and their limitations discussed. Particular emphasis is placed on the charge distributions and interfacial potential profiles expected at various types of electroactive interfaces. PMID- 15117255 TI - Semiclassical description of molecular dynamics based on initial-value representation methods. AB - Recent progress in the development of semiclassical methods to describe quantum effects in molecular dynamics is reviewed. Focusing on rigorous semiclassical methods that are based on the initial-value representation of the semiclassical propagator, we discuss several promising schemes that have been developed in the past few years to extend the applicability of semiclassical approaches to complex molecular systems. In particular, integral-filtering techniques and forward backward methods are surveyed. Furthermore, recently proposed approaches that allow the semiclassical description of nonadiabatic molecular dynamics are discussed. The potential and efficiency of these methods is illustrated by selected applications. PMID- 15117256 TI - Quantitative prediction of crystal-nucleation rates for spherical colloids: a computational approach. AB - This review discusses the recent progress that has been made in the application of computer simulations to study crystal nucleation in colloidal systems. We discuss the concept and the numerical methods that allow for a quantitative prediction of crystal-nucleation rates. The computed nucleation rates are predicted from first principles and can be directly compared with experiments. These techniques have been applied to study crystal nucleation in hard-sphere colloids, polydisperse hard-sphere colloids, weakly charged or slightly soft colloids, and hard-sphere colloids that are confined between two-plane hard walls. PMID- 15117257 TI - Proton-coupled electron transfer: a reaction chemist's view. AB - Proton-coupled electron transfer (PCET) reactions involve the concerted transfer of an electron and a proton. Such reactions play an important role in many areas of chemistry and biology. Concerted PCET is thermochemically more favorable than the first step in competing consecutive processes involving stepwise electron transfer (ET) and proton transfer (PT), often by >=1 eV. PCET reactions of the form X-H + Y X + H-Y can be termed hydrogen atom transfer (HAT). Another PCET class involves outersphere electron transfer concerted with deprotonation by another reagent, Y+ + XH-B Y + X-HB+. Many PCET/HAT rate constants are predicted well by the Marcus cross relation. The cross-relation calculation uses rate constants for self-exchange reactions to provide information on intrinsic barriers. Intrinsic barriers for PCET can be comparable to or larger than those for ET. These properties are discussed in light of recent theoretical treatments of PCET. PMID- 15117258 TI - Neutron reflection from liquid interfaces. AB - Recent applications of neutron reflectometry to the study of wet interfaces are described. An outline is given of the basic principles that allow the techniques to determine composition and structure in a variety of situations. These are the adsorption of surfactant molecules at air/liquid and solid/liquid interfaces, the shape of the segment-density profiles of different types of polymer, including block copolymers and polyelectrolytes, adsorption in mixed surfactant and polymer/surfactant systems, and interfacial systems of biophysical interest. PMID- 15117259 TI - Time-dependent density functional theory. AB - Time-dependent density functional theory (TDDFT) can be viewed as an exact reformulation of time-dependent quantum mechanics, where the fundamental variable is no longer the many-body wave function but the density. This time-dependent density is determined by solving an auxiliary set of noninteracting Schrodinger equations, the Kohn-Sham equations. The nontrivial part of the many-body interaction is contained in the so-called exchange-correlation potential, for which reasonably good approximations exist. Within TDDFT two regimes can be distinguished: (a) If the external time-dependent potential is "small," the complete numerical solution of the time-dependent Kohn-Sham equations can be avoided by the use of linear response theory. This is the case, e.g., for the calculation of photoabsorption spectra. (b) For a "strong" external potential, a full solution of the time-dependent Kohn-Sham equations is in order. This situation is encountered, for instance, when matter interacts with intense laser fields. In this review we give an overview of TDDFT from its theoretical foundations to several applications both in the linear and in the nonlinear regime. PMID- 15117260 TI - Theory of single-molecule spectroscopy: beyond the ensemble average. AB - Single-molecule spectroscopy (SMS) is a powerful experimental technique used to investigate a wide range of physical, chemical, and biophysical phenomena. The merit of SMS is that it does not require ensemble averaging, which is found in standard spectroscopic techniques. Thus SMS yields insight into complex fluctuation phenomena that cannot be observed using standard ensemble techniques. We investigate theoretical aspects of SMS, emphasizing (a) dynamical fluctuations (e.g., spectral diffusion, photon-counting statistics, antibunching, quantum jumps, triplet blinking, and nonergodic blinking) and (b) single-molecule fluctuations in disordered systems, specifically distribution of line shapes of single molecules in low-temperature glasses. Special emphasis is given to single molecule systems that reveal surprising connections to Levy statistics (i.e., blinking of quantum dots and single molecules in glasses). We compare theory with experiment and mention open problems. Our work demonstrates that the theory of SMS is a complementary field of research for describing optical spectroscopy in the condensed phase. PMID- 15117261 TI - Optically detected magnetic resonance studies of colloidal semiconductor nanocrystals. AB - The review describes the studies of the magneto-optical properties of II-VI and III-V semiconductor nanocrystals (NCs) capped with organic or inorganic epitaxial shells. The investigations focused on the chemical identification of localization sites (core, shell, or interface) of photogenerated carriers in spherical NCs and elucidated the influence of the surface/interface quality on the optical properties of the materials. Optically detected magnetic resonance (ODMR) spectroscopy was used for the study of the proposed physical properties. The ODMR method provides the means to identify the surface/interface sites and correlate them with specific optical transition. In addition, this method reveals information about the spin multiplicity of band edge and trapped states and the electron-hole exchange interaction, determines the spectroscopic g-factors, distinguishes between the radiative and nonradiative characteristic of a trapping site, and evaluates the spin-lattice relaxation times. PMID- 15117262 TI - Amorphous water. AB - After providing some background material to establish the interest content of this subject, we summarize the many different ways in which water can be prepared in the amorphous state, making clear that there seems to be more than one distinct amorphous state to be considered. We then give some space to structural and spectroscopic characterization of the distinct states, recognizing that whereas there seems to be unambiguously two distinct states, there may be in fact be more, the additional states mimicking the structures of the higher-density crystalline polymorphs. The low-frequency vibrational properties of the amorphous solid states are then examined in some detail because of the gathering evidence that glassy water, while difficult to form directly from the liquid like other glasses, may have some unusual and almost ideal glassy features, manifested by unusually low states of disorder. This notion is pursued in the following section dealing with thermodynamic and relaxational properties, where the uniquely low excess entropy of the vitreous state of water is confirmed by three different estimates. The fact that the most nearly ideal glass known has no properly established glass transition temperature is highlighted, using known dielectric loss data for amorphous solid water (ASW) and relevant molecular glasses. Finally, the polyamorphism of glassy water, and the kinetic aspects of transformation from one form to the other, are reviewed. PMID- 15117263 TI - Single-molecule optics. AB - We review recent developments in single-molecule spectroscopy and microscopy. New optical methods provide access to the absorption, emission, or excitation spectra of single nano-objects and can determine either the positions of these objects with subwavelength accuracy or the full three-dimensional orientation of their transition dipole moments. Recent work aims at using single molecules as nanoparts or nanoelements in a variety of molecular-scale devices, from triggered sources of single photons to single-molecular switches. A prominent new direction explores the various interactions between molecules within individual multichromophoric systems obtained by chemical synthesis. These systems are the models for natural self-assembled systems such as the light-harvesting proteins of bacteria and green plants, which are currently studied on a single-molecule basis. Another important class of multichromophoric systems are conjugated polymers. The combination of microscopy with time- and frequency-resolved spectroscopy is opening a wide field of new and exciting applications to individual nano-objects. PMID- 15117264 TI - Biomimetic nanoscale reactors and networks. AB - Methods based on self-assembly, self-organization, and forced shape transformations to form synthetic or semisynthetic enclosed lipid bilayer structures with several properties similar to biological nanocompartments are reviewed. The procedures offer unconventional micro- and nanofabrication routes to yield complex soft-matter devices for a variety of applications for example, in physical chemistry and nanotechnology. In particular, we describe novel micromanipulation methods for producing fluid-state lipid bilayer networks of nanotubes and surface-immobilized vesicles with controlled geometry, topology, membrane composition, and interior contents. Mass transport in nanotubes and materials exchange, for example, between conjugated containers, can be controlled by creating a surface tension gradient that gives rise to a moving boundary or by induced shape transformations. The network devices can operate with extremely small volume elements and low mass, to the limit of single molecules and particles at a length scale where a continuum mechanics approximation may break down. Thus, we also describe some concepts of anomalous fluctuation-dominated kinetics and anomalous diffusive behaviours, including hindered transport, as they might become important in studying chemistry and transport phenomena in these confined systems. The networks are suitable for initiating and controlling chemical reactions in confined biomimetic compartments for rationalizing, for example, enzyme behaviors, as well as for applications in nanofluidics, bioanalytical devices, and to construct computational and complex sensor systems with operations building on chemical kinetics, coupled reactions and controlled mass transport. PMID- 15117284 TI - Don't call me nuts: an international perspective on the stigma of mental illness. PMID- 15117283 TI - Domain structure of bi-functional selenoprotein P. AB - Human selenoprotein P (SeP), a selenium-rich plasma glycoprotein, is presumed to contain ten selenocysteine residues; one of which is located at the 40th residue in the N-terminal region and the remaining nine localized in the C-terminal third part. We have shown that SeP not only catalyses the reduction of phosphatidylcholine hydroperoxide by glutathione [Saito, Hayashi, Tanaka, Watanabe, Suzuki, Saito and Takahashi (1999) J. Biol. Chem. 274, 2866-2871], but also supplies its selenium to proliferating cells [Saito and Takahashi (2002) Eur. J. Biochem. 269, 5746-5751]. Treatment of SeP with plasma kallikrein resulted in a sequential limited proteolysis (Arg-235-Gln-236 and Arg-242-Asp 243). The N-terminal (residues 1-235) and C-terminal (residues 243-361) fragments exhibited enzyme activity and selenium-supply activity respectively. These results confirm that SeP is a bi-functional protein and suggest that the first selenocysteine residue is the active site of the enzyme and the remaining nine residues function as a selenium supplier. PMID- 15117285 TI - Novel antipsychotics in bipolar and schizoaffective mania. AB - OBJECTIVE: Novel antipsychotics are increasingly used in the treatment of bipolar and schizoaffective mania. This paper presents an overview of the controlled studies in this field. METHOD: Using cross-references, a computerized search was performed on MEDLINE and EMBASE psychiatry covering the period 1990-2002. RESULTS: Olanzapine and risperidone, added to mood stabilizers, and olanzapine as monotherapy enjoy the most evidential support in terms of efficacy and side effect profile for their use in acute bipolar mania. The use of modern antipsychotics in bipolar prophylaxis and in both the short- and long-term treatment of schizomania has not been widely studied yet. CONCLUSION: More controlled trials are still needed comparing modern antipsychotics as monotherapy and adjunctive to mood stabilizers with conventional antipsychotics, lithium, anticonvulsants and with each other in short-term and, especially, maintenance treatment of (schizo)mania. Partly based on controlled studies, olanzapine, risperidone and other modern antipsychotics could become preferable for these indications. PMID- 15117286 TI - Effects of labelling on public attitudes towards people with schizophrenia: are there cultural differences? AB - OBJECTIVE: A representative survey which was recently conducted in Germany came to the conclusion that labelling as mental illness has an impact on public attitudes towards people with schizophrenia, with negative effects clearly outweighing positive effects. In this study, we will examine whether this result can be replicated in other countries. METHOD: In the summer of 2002, representative surveys were carried out in Novosibirsk (Russia) and in Ulaanbaatar (Mongolia), using the same sampling procedure and interview as in Germany. RESULTS: As in Germany, in Novosibirsk and Ulaanbaatar labelling as mental illness was positively correlated with the endorsement of the belief that the individual depicted in the vignette is in need for help. However, unlike in Germany, labelling had no significant effect on the endorsement of the stereotype of dangerousness. CONCLUSION: Our findings support the notion that labelling effects are culture-related. Therefore, anti-stigma efforts need to be tailored to the specific conditions in a particular country. PMID- 15117287 TI - Pretreatment attrition and dropout in an outpatient clinic for anxiety disorders. AB - OBJECTIVE: Treatment attrition represents a considerable problem for efficient delivery of care for mental disorders. The present study examined rates and predictors of pretreatment attrition and dropout from outpatient treatment for anxiety disorders. METHOD: The influence of clinical, demographic, clinician and system variables on pretreatment attrition (treatment refusal or non-attendance) and dropout were analysed in a consecutive sample of 731 clients treated at an anxiety disorders clinic in Sydney. RESULTS: Pretreatment attrition was common (30.4%) but dropout once clients had commenced treatment was rare (10.3%). Milder pretreatment symptoms were associated with treatment dropout and comorbid depression or depressive symptoms were associated with higher probability of both pretreatment attrition and dropout. Demographic, clinician and system influences were present but less important. CONCLUSION: Targeting of individuals with comorbid anxiety and depression may reduce treatment attrition rates among individuals with anxiety disorders presenting for treatment. PMID- 15117288 TI - Outcome of postpartum disorders: a 10 year follow-up of hospital admissions. AB - OBJECTIVE: To determine readmission and recurrence rates of women admitted to psychiatric hospital with their babies, and index factors predicting subsequent course. METHOD: A total of 66 women were followed up after 10 years. Information was obtained from structured interviews or other personal contact, and from general practitioners and hospital notes. Psychiatric diagnoses were made according to Research Diagnostic Criteria (RDC). Outcome and predictive factors were investigated using survival analysis. RESULTS: A recurrence rate of 87.2%, and a readmission rate of 63.3% were found. Readmissions were more common after index schizophrenia than other diagnoses. The strongest predictor of readmission was previous psychiatric history. Among 27 subsequent pregnancies, puerperal recurrence was high among those with index psychoses (75-80%), but lower among those with index depression (27.3%). CONCLUSION: The recurrence rate is high, with a moderate readmission rate predicted by past psychiatric history. Continuing clinical vigilance over the long-term is required for these patients. PMID- 15117289 TI - Psychosocial predictors of successful delivery after unexplained recurrent spontaneous abortions: a cohort study. AB - OBJECTIVE: To examine psychosocial predictors of successful pregnancy after recurrent spontaneous abortions (RSA). METHOD: We administered two waves of semi structured interview and self-report questionnaire battery to a consecutive series of 46 couples who had had two RSAs (baseline before third pregnancy, and immediately after third pregnancy was ascertained), and followed them through their third pregnancy. RESULTS: Of the 46 couples, four miscarried for karyotypal abnormalities and six without any known cause. When the latter six were compared with the remaining 36 women, they reported less social support satisfaction, a more stable attribution for the causes of past abortions, and more depressed mood in the preceding year. Taken altogether, this psychosocial model was able to predict 93% of the pregnancy outcomes correctly. CONCLUSION: Psychosocial variables (depression, attribution and social support) are robust predictors of the prospective pregnancy and suggest possible points of intervention in couples with RSA. PMID- 15117290 TI - Attachment security and parental bonding in adults with obsessive-compulsive disorder: a comparison with depressed out-patients and healthy controls. AB - OBJECTIVE: This study examines concurrent associations of attachment security, psychopathology and recollections of early parental interactions, in adults with obsessive-compulsive disorder (OCD), depression, and in healthy controls. METHOD: Thirty-six out-patients with OCD, 16 depressed out-patients and 26 controls were asked to fill out the Revised Adult Attachment Scale and the Parental Bonding Instrument (PBI). RESULTS: OCD and depressed groups were more insecure than controls. The depressed group recalled less caring mothers than the OCD group, while the OCD group was indistinguishable from controls on PBI measures. Married status was associated with greater security, but also with recollections of greater parental control, and lower maternal care. CONCLUSION: OCD and depressed groups demonstrated greater attachment insecurity than controls. No clear relationship emerged between security and PBI recollections. The PBI may not measure aspects of early interactions essential for later attachment security, or recollections may be biased according to diagnosis or attachment style. PMID- 15117291 TI - Mood changes during pregnancy and the postpartum period: development of a biopsychosocial model. AB - OBJECTIVE: Women are vulnerable to mood changes during pregnancy and the postpartum period. We set out to empirically test the hypothesis that biological and psychosocial variables interact to result in this vulnerability. METHOD: Using structural equation modeling techniques, we developed an integrative model of perinatal mood changes from clinical, psychosocial, hormone and mood data collected from 150 women in late pregnancy and at 6-weeks postpartum. RESULTS: In the prenatal model, biological variables had no direct effect on depressive symptoms. However, they did act indirectly through their significant effects on psychosocial stressors and symptoms of anxiety. The same model did not fit the postpartum data, suggesting that different causal variables may be implicated in postpartum mood. CONCLUSION: This model demonstrates the importance of considering both biological and psychosocial variables in complex health conditions such as perinatal mood disorders. PMID- 15117296 TI - Treating depression in the developing world. PMID- 15117297 TI - The effectiveness of active population screening and treatment for sleeping sickness control in the Democratic Republic of Congo. AB - BACKGROUND: The human African trypanosomiasis (HAT) control programme of the Democratic Republic of Congo (DRC) uses mass screening with the card agglutination test for trypanosomes (CATT). We looked at the contribution of CATT and improved parasitological confirmation to the effectiveness of screening and treatment. METHOD: The effectiveness of the screening and treatment process is measured by the percentage of HAT cases that is effectively cured after a single round of screening. The process is analysed in five steps: (i) the attendance at the screening, (ii) the sensitivity of the screening procedure, (iii) the sensitivity of the parasitological confirmation, (iv) the proportion of the confirmed cases that effectively receive treatment and (v) the cure rate of the treatment. We used a simplified model that multiplies proportions of infected persons that go through each step. We estimated these parameters using a combination of routine data collected by the national control programme over the period January 1997 to December 1998 and published data. For varying attendance rates we compared the effectiveness of screening strategies based on CATT or on CATT combined with improved parasitological confirmation by mini anion exchange column technique (mAECT) with the previously used strategy based on palpation of neck glands and microscopy alone. RESULTS: The model shows that overall effectiveness of the active case detection and treatment strategy is <50% under most scenarios. Attendance rates averaged 74% but showed considerable regional variability and are a major problem in some areas of DRC. The CATT and replacing traditional parasitology by mAECT increases the sensitivity of the screening but a substantial part of the gains are lost at other stages of the screening process. CONCLUSION: Improvements of the HAT screening process such as introduction of CATT or mAECT only make sense if other parameters and attendance rate in particular are optimized at the same time. PMID- 15117298 TI - Split-drug regimens for the treatment of patients with sputum smear-positive pulmonary tuberculosis--a unique approach. AB - OBJECTIVE: To evaluate the efficacy of split-drug regimens for treatment of patients with sputum smear-positive pulmonary tuberculosis in south India. DESIGN: Randomized controlled clinical trial where eligible patients were randomly allocated to: (i) 2RE(3)HZ(3)(alt)/4RH(2) (split I): rifampicin plus ethambutol given on one day and isoniazid plus pyrazinamide the next day for first 2 months followed by rifampicin plus isoniazid twice weekly for 4 months, or (ii) 3RE(3)HZ(3)(alt)/3RH(2) (split II): similar to regimen 1, except duration was 3 months in each phase, or (iii) 2REHZ(3)/4RH(2) (control): rifampicin, isoniazid, ethambutol and pyrazinamide, given thrice weekly for 2 months followed by isoniazid and rifampicin twice weekly for 4 months. All patients were followed up clinically and bacteriologically every month up to 2 years and every 6 months for up to 5 years. RESULTS: A favourable response (cultures negative for Mycobacterium tuberculosis during the last 2 months of treatment) was observed in 91% of 407 patients in split I, 94% of 415 in split II and 89% of 418 in the control regimen. Ninety-one per cent of 370 patients in split I, 93% of 389 in split II and 90% of 370 in control regimens had quiescent disease at the end of 60 months. Gastrointestinal symptoms were more frequent under the control regimen (P = 0.01). CONCLUSION: Split-drug regimens were as effective as the control regimen in terms of favourable response at the end of treatment and quiescent disease at 5 years, and caused fewer gastrointestinal side-effects. PMID- 15117299 TI - Direct observation of treatment for tuberculosis: a randomized controlled trial of community health workers versus family members. AB - We implemented community-based direct observation of treatment, short course (DOTS), including a randomized controlled trial of direct observation either by community health workers (CHWs) or family members, under operational conditions in a region of Swaziland. There was a high death rate of 15%, due to the high HIV rates in the region. There was no significant difference in the cure and completion rate between direct observation of treatment by CHWs and family members [2% difference (95% CI -3% to 7%), exact P = 0.52]. A before-and-after comparison of outcomes demonstrated that the cure and treatment completion rate improved from a baseline of 27-67% following implementation of community-based DOTS. We conclude that community-based tuberculosis DOTS can improve successful outcomes of treatment. However, direct observation can be undertaken effectively using either daily family or CHW supervision. The choice of treatment supporter should be based on access, patient preference and availability of CHW resource. PMID- 15117300 TI - A randomized trial on acceptability of voluntary HIV counselling and testing. AB - OBJECTIVES: To examine factors affecting readiness for and acceptability of voluntary HIV counselling and testing (VCT). METHODS: Participants in a population-based HIV survey conducted in an urban population in Zambia in 1996 were offered VCT. Although 29% of them expressed interest in being tested (readiness), only 4% of this group used the services (i.e. acceptability). When the survey was repeated 3 years later, VCT was designed differently to assess acceptability. At the cluster level the participants were randomly allocated to VCT either at the local clinic (similar to 1996, n = 1102) or at an optional location (n = 1343). RESULTS: Readiness varied significantly by age group (47% in age group 20-24 years vs. 18% in age group 40-49 years). There were contrasts between young (15-24 years) and older age groups (25-49 years) regarding the main factors associated with readiness. Whereas self-perceived risk of being HIV infected was the only significant factor among the young, poor self-rated health and ever HIV tested were important factors among the older. The acceptability was 11.8% among the group allocated to VCT at the local clinic compared with 55.8% for the group allocated to an optional location (RR, 4.7). CONCLUSIONS: Perceived risk of HIV infection had a major influence on VCT readiness among young people, whereas declining general health status, as indicated by self-rated health, was most evident among those of older age. A strong effect of placement on acceptability of VCT was demonstrated, indicating this barrier to be important in explaining low demands for VCT in the past. Differences in perceptions of how confidentiality is handled at the two locations might be an important underlying factor. PMID- 15117301 TI - Evaluation of an algorithm for integrated management of childhood illness in an area of Vietnam with dengue transmission. AB - OBJECTIVES: To determine whether nurses, using the WHO/UNICEF algorithm for integrated management of childhood illness (IMCI), modified to include dengue infection, satisfactorily classified children in an area endemic for dengue haemorrhagic fever (DHF). METHODS: Nurses assessed and classified, using the modified IMCI algorithm, a systematic sample of 1250 children aged 2 months to 10 years (n = 1250) presenting to a paediatric hospital in Dong Nai Province, Vietnam. Their classification was compared with that of a paediatrician, blind to the result of the nurses' assessment, which could be modified in the light of simple investigations, e.g. dengue serology. RESULTS: In children aged 2-59 months (n = 859), the nurses were able to classify, using the modified chart, the presenting illness in >99% of children and found more than one classification in 70%. For the children with pneumonia, diarrhoea, dengue shock syndrome, severe DHF and severe disease requiring urgent admission, the nurse's classification was >60% sensitive and >85% specific compared with that of the paediatrician. For the nurse's classification of DHF the specificity was 50-55% for the children <5 years and in children with definitive dengue serology. Alterations in the DHF algorithm improved specificity at the expense of sensitivity. CONCLUSION: Using the IMCI chart, nurses classified appropriately many of the major clinical problems in sick children <5 years in southern Vietnam. However, further modifications will be required in the fever section, particularly for dengue. The impact of using the IMCI chart in peripheral health stations remains to be evaluated. PMID- 15117302 TI - Genetic heterogeneity of the dengue vector Aedes aegypti in Martinique. AB - In Martinique, Aedes aegypti, the vector of dengue viruses has been the target of insecticide control for more than 35 years. Despite significant control efforts, dengue has become a major disease of public health concern. We conducted a population genetic analysis based on isoenzyme variations combined with an estimation of infection rate to a dengue virus among 26 Ae. aegypti samples. Aedes aegypti samples could be differentiated for their susceptibility to dengue infection (infection rates ranging from 42.8% to 98.6%) and showed important genetic variation (significant F(ST) values). PMID- 15117303 TI - Prevalence of Opisthorchis viverrini infection and incidence of cholangiocarcinoma in Khon Kaen, Northeast Thailand. AB - Liver cancer is the most common cancer in Khon Kaen, Northeast Thailand, because of the high incidence of cholangiocarcinoma (CHCA). Opisthorchis viverrini (OV), a liver fluke, is endemic in the area, and has been evaluated as a cause of CHCA by International Agency for Research on Cancer. Residents of 20 districts in the province were invited to attend a mobile screening programme between 1990 and 2001. Of 24 723 participants, 18 393 aged 35-69 years were tested for OV infection, by examining stools for the presence of eggs. Prevalence of infection in each district was estimated from the sample of the population who had been tested. The incidence of liver cancer in 1990-2001 was obtained for each district from the cancer registry. The average crude prevalence of OV infection in the sample subjects was 24.5%, ranging from 2.1% to 70.8% in different districts. Truncated age-standardized incidence of CHCA at ages >35 years varied threefold between districts, from 93.8 to 317.6 per 100,000 person-years. After adjustment for age group, sex and period of sampling, there was a positive association between prevalence of OV infection and incidence of CHCA at the population level. Associations between CHCA and active OV infection in individuals have become hard to demonstrate, because of effective anti-OV treatment. The relationship may, however, be clear in comparisons between populations, which, for infectious diseases, take into account the contextual effects of group exposure in determining individual outcome. The cancer registry is an appropriate tool for disease monitoring in small areas. PMID- 15117304 TI - High prevalence of ectopic kidney in Coast Province, Kenya. AB - OBJECTIVE: To establish the prevalence of congenital urinary tract abnormalities in a full population-based ultrasound survey of an area of coastal Kenya. METHODS: Ultrasound examination of 3118 residents of 912 households, including all available subjects over 2 years of age, residing in five contiguous rural villages 50 km south of Mombasa. RESULTS: Survey findings indicated simple renal ectopia in 11 of 3118 subjects (0.35%) and renal agenesis in three (0.096%). No cases of horseshoe kidney or complex urinary anomaly were detected, and no cases of multiple congenital anomaly were found. Ectopia cases were evenly distributed between men and women, and across the five study villages. None of the individuals affected by renal ectopia were closely related (i.e. <5th-degree relations). CONCLUSION: There is an unusually high prevalence of ectopia among unrelated subjects in this area. In this setting, the findings suggest either a common exposure to teratogenetic factors, or a hereditary condition with variable penetrance, where more severely affected individuals are not observed because of foetal/infant mortality. PMID- 15117305 TI - Evidence for increased metabolism of chloroquine during the early third trimester of human pregnancy. AB - OBJECTIVE: To examine the possibility of a different extent of chloroquine (CQ) metabolism in human pregnancy by determining blood level profiles of the drug and its major metabolite, desethylchloroquine (CQM). METHODS: Five women in the early third trimester of pregnancy and five non-pregnant women received each a single 600 mg oral dose of CQ and blood samples were collected at pre-determined intervals following drug administration. Plasma concentrations of CQ and CQM were analysed by an established HPLC method. RESULTS: The C(max) and AUC(0-48 h) of CQM were significantly higher in the pregnant than the non-pregnant group (P = 0.009). The ratio AUC(CQ)/AUC(CQM) ranged from 0.09 to 0.35 among pregnant women, and from 1.70 to 4.81 among non-pregnant women. CONCLUSION: Results from this preliminary study indicate an occurrence of induction of metabolism of CQ in the early third trimester of pregnancy. In view of toxicological importance of CQ metabolites, it is suggested that caution should be exercised in evaluation of higher dosage regimen of CQ in pregnant women. PMID- 15117306 TI - Open randomized study of pyrimethamine-sulphadoxine vs. pyrimethamine sulphadoxine plus probenecid for the treatment of uncomplicated Plasmodium falciparum malaria in children. AB - BACKGROUND: Increasing drug resistance in Plasmodium falciparum has necessitated renewed search for cheap, effective alternatives to commonly available antimalarials, chloroquine and pyrimethamine-sulphadoxine, for the treatment of malaria in Africa. Probenecid, an inhibitor of organic anion transporters and multiresistance-associated proteins, can chemosensitize P. falciparum to pyrimethamine and sulphadoxine in vitro, but the clinical significance is unclear. We assessed the safety, treatment efficacy, and effects on gametocyte carriage of adding probenecid to pyrimethamine-sulphadoxine. METHODS: We evaluated 151 children aged 12 years or younger who had uncomplicated P. falciparum malaria. Patients were randomly assigned pyrimethamine-sulphadoxine (25 mg/kg of the sulphadoxine component) or pyrimethamine-sulphadoxine as above plus probenecid 20-25 mg/kg of bodyweight in two divided doses daily for 3 days. The primary endpoints were parasitological cure rates on days 14 and 28. RESULTS: Both regimens were well tolerated; no child was withdrawn because of drug intolerance. Fever (1.9 +/- 1.1 vs. 2.4 +/- 1.2 days, P = 0.02) and parasite clearance (2.3 +/- 0.9 vs. 2.7 +/- 1.1 days, P = 0.04) were significantly shorter, and the parasitological cure rate on day 14 (96.2%vs. 83.5%, P = 0.02) but not day 28 (79.4%vs. 72.6%, P = 0.4), was significantly higher in children treated with pyrimethamine-sulphadoxine-probenecid than in those treated with pyrimethamine-sulphadoxine. Gametocyte carriage was similar with both treatment regimens. CONCLUSIONS: The combination of pyrimethamine-sulphadoxine, and probenecid, at a relatively moderate dose, improved treatment efficacy but had no effect on gametocyte carriage. The pyrimethamine-sulphadoxine-probenecid combination merits further evaluation as a potential treatment for use in Nigeria. PMID- 15117307 TI - Rapid epidemiologic assessment of glucose-6-phosphate dehydrogenase deficiency in malaria-endemic areas in Southeast Asia using a novel diagnostic kit. AB - We recently reported a new rapid screening method for glucose-6-phosphate dehydrogenase (G6PD) deficiency. This method incorporates a new formazan substrate (WST-8) and is capable of detecting heterozygous females both qualitatively and quantitatively. Here, we report its evaluation during field surveys at three malaria centres and in malaria-endemic villages of Myanmar and Indonesia, either alone or in combination with a rapid on-site diagnosis of malaria. A total of 57 severe (45 males and 12 females) and 34 mild (five males and 29 females) cases of G6PD deficiency were detected among 855 subjects in Myanmar whilst 30 severe (25 males and five females) and 23 mild (six males and 17 females) cases were found among 1286 subjects in Indonesia. In all cases, severe deficiency was confirmed with another formazan method but due to limitations in its detection threshold, mild cases were misdiagnosed as G6PD normal by this latter method. Our results indicate that the novel method can qualitatively detect both severely deficient subjects as well as heterozygous females in the field. The antimalarial drug, primaquine, was safely prescribed to Plasmodium vivax-infected patients in Myanmar. Our new, rapid screening method may be essential for the diagnosis of G6PD deficiency particularly in rural areas without electricity, and can be recommended for use in malaria control programmes. PMID- 15117308 TI - Relationship between age, molecular markers, and response to sulphadoxine pyrimethamine treatment in Kampala, Uganda. AB - Sulphadoxine/pyrimethamine (SP) has become the first-line treatment of uncomplicated malaria in a number of African countries. Molecular surveillance of resistance-mediating mutations in Plasmodium falciparum dihydrofolate reductase (dhfr) and dihydropteroate synthase (dhps) has been proposed as a means of predicting SP treatment outcomes, but optimal methods of surveillance in different populations have not been well established. To investigate the relationship between molecular markers of SP resistance, host immunity, and response to therapy, we evaluated the association between the presence of five key dhfr and dhps mutations at enrollment and clinical outcome in children and adults treated with SP for uncomplicated malaria in Kampala, Uganda. Clinical treatment failure was 11% at 14 days, increasing to 30% at 28 days, after excluding new infections. Outcomes varied markedly based on the number of dhfr and dhps mutations and on the age of treated subjects. All infections with less than two dhfr/dhps mutations were successfully treated. Treatment failure associated with any two, three, or four dhfr/dhps mutations occurred in nine of 24 (38%) children up to 5 years, but not in older patients (0/20). In the presence of all five mutations, treatment failure occurred equally in children aged 5 years or younger [7/16 (44%)] and in older patients [8/16 (50%)]. Our results showed that age, a surrogate marker of antimalarial immunity, had a major impact on the relationship between polymorphisms in SP target enzymes and treatment outcomes. The use of molecular markers of SP resistance to predict treatment failure rates should take age into account. PMID- 15117309 TI - Implementation of intermittent preventive treatment with sulphadoxine pyrimethamine for control of malaria in pregnancy in Kisumu, western Kenya. AB - OBJECTIVE: In 1998, the Kenyan Ministry of Health introduced intermittent preventive treatment (IPT) with sulphadoxine-pyrimethamine (SP), one treatment dose in the second trimester (16-27 weeks) and one treatment dose between 28 and 34 weeks of gestational age, for the control of malaria in pregnancy. We evaluated the coverage and determinants of receipt of IPT after its introduction in the Provincial Hospital in Kisumu, western Kenya. METHODS: Information on the use of IPT in pregnancy was collected from women who attended the antenatal clinic (ANC) and delivered in the same hospital. In exit interviews, we assessed patterns of IPT use in the ANC. RESULTS: Of 1498 women who delivered between June 1999 and June 2000, 23.7%, 43.4% and 32.9% received > or =2, 1 or no dose of SP, respectively. Late first ANC attendance was the most important factor contributing to incomplete IPT; 45% of the women started attending ANC in the third trimester. More women received at least one tetanus toxoid immunization than at least one dose of IPT (94%vs. 67%, P < 0.05). In exit interviews, 74% correctly associated IPT with treatment of malaria; however, knowledge on the need for the second dose was poor. Three per cent of the administrations were given despite contraindications. The agreement between gestational age by date of last menstrual period and by palpation was low (kappa = 0.1). CONCLUSIONS: Education of pregnant women and ANC staff to increase earlier attendance for ANC has the potential to substantially increase the proportion of women receiving two doses of IPT with SP. PMID- 15117310 TI - Quality of sexually transmitted infections services for female sex workers in Abidjan, Cote d'Ivoire. AB - OBJECTIVES: To assess the quality of sexually transmitted infections (STI) care in health care facilities in Abidjan attended by female sex workers. METHODS: A cross-sectional study was conducted in June 2000 in the 29 health care facilities and 10 pharmacies, which were reported as points of first encounter for STI care by female sex workers in a previous study on health seeking behaviour. Evaluation components included: (1) checklists of equipment and STI drugs in the facilities; (2) interviews with health care providers and pharmacists; (3) direct observation of the provider/client interaction; (4) exit interviews with women attending with STI or genital problems. RESULTS: Private health care facilities were more expensive, had fewer clients, and had less equipment and medical staff than public facilities, with the exception of the special female sex worker clinic. A total of 60 health care providers and 29 pharmacists were interviewed. There was no difference in their scoring on syndromic approach case studies, with the exception of the nurse assistants, who scored less. Overall scores for correct treatment were lowest for the pharmacists. We observed 513 provider-client interactions, of which 161 related to STIs or genital problems in women. Questions about recent sexual contacts were asked in only 20% and preventive messages were given in only 9% of the cases with STI/genital problems. Of 161 clients interviewed, 44% complained about a long waiting time, and 39% thought the health care provider had adequately explained the problem to them. CONCLUSIONS: The opportunity for improvement of STI case management in health care facilities in Abidjan where female sex workers go for STI care is enormous. Public and private health care facilities should be made more accessible for sex workers, and their services should be upgraded to better respond to the sexual health needs of high risk women. PMID- 15117311 TI - Maternal perceptions of acute respiratory infections in children under 5 in rural South Africa. AB - OBJECTIVES: To assess maternal ability to recognize respiratory distress and to identify local beliefs and practices around respiratory infections in rural KwaZulu/Natal, South Africa. METHODS: A descriptive study: 15 knowledgeable mothers were shown a video of 10 children with respiratory distress and four normal children. Mothers were asked to describe perceived types, signs, symptoms, causes of and actions taken for each child. Sensitivity and specificity were calculated for maternal recognition of respiratory distress (chest indrawing, fast breathing) shown on the video. A focus group discussion with six mothers was held to corroborate and refine individually reported local concepts. FINDINGS: Maternal recognition of respiratory distress was good (sensitivity 91.3%, 95% CI: 86.8-95.8%; specificity 95%, 95%CI: 89.5-100%), with little variation between mothers (kappa = 0.704). Mothers described 12 'local types of respiratory illness', each with its own name and its own well-defined set of signs and symptoms. They were classifiable into five causative categories: supernatural, natural, tuberculosis, cold weather and unknown, indicating that perceptions of causation differed greatly from biomedical concepts. For illnesses of perceived supernatural causation, mothers were reluctant to seek medical care and antibiotics were deemed inappropriate. Traditional remedies were preferred instead. CONCLUSION: Knowledge of local vocabulary and concepts about respiratory infections is essential to design strategies for health care workers to communicate with mothers about respiratory disease, so that mothers will seek timely medical care, provide continued safe, supportive home care and comply with antibiotic treatment. PMID- 15117312 TI - Short communication: Misleading microscopy in amoebiasis. AB - High prevalences of intestinal amoebiasis are commonly reported by microscopy in Ethiopia. In order to confirm the actual occurrence of Entamoeba histolytica we collected 108 stool specimens from different hospitals & health centers from patients in whom haematophagous trophozoites were believed to be found. We detected only a single E. histolytica case while 77 (71.3%) were E. dispar and the remaining 30 samples were negative for both species by real-time PCR based on the small subunit ribosomal RNA gene sequence of E. histolytica and E. dispar. The tradition of microscopy in a routine diagnostic set-up appears unsatisfactory to reliably differentiate rbc-engulfing amoeba from non-invasive amoeba in wet smears. PMID- 15117313 TI - Rab GTPases and myosin motors in organelle motility. AB - The actin cytoskeleton is essential to ensure the proper location of, and communication between, intracellular organelles. Some actin-based myosin motors have been implicated in this process, particularly members of the class V myosins. We discuss here the emerging role of the Ras-like GTPases of the Rab family as regulators of myosin function in organelle transport. Evidence from yeast secretory vesicles and mitochondria, and mammalian melanosomes and endosomes suggests that Rab GTPases are crucial components of the myosin organelle receptor machinery. Better understood is the case of the melanosome where Rab27a recruits a specific effector called melanophilin, which in turn binds myosin Va. The presence of a linker protein between a Rab and a myosin may represent a general mechanism. We argue that Rabs are ideally suited to perform this role as they are exquisite organelle markers. Furthermore, the molecular switch property of Rabs may enable them to regulate the timing of the myosin association with the target organelle. PMID- 15117314 TI - N-terminal kinesins: many and various. AB - Molecular motors are a fascinating group of proteins that have vital roles in a huge variety of cellular processes. They all share the ability to produce force through the hydrolysis of adenosine triphosphate, and fall into classes groups: the kinesins, myosins and the dyneins. The kinesin superfamily itself can be split into three major groups depending on the position of the motor domain, which is localized N-terminally, C-terminally, or internally. This review focuses on the N-terminal kinesins, providing a brief overview of their roles within the cell, and illustrating recent key developments in our understanding of how these proteins function. PMID- 15117315 TI - VisBio: a computational tool for visualization of multidimensional biological image data. AB - New laser scanning microscopy techniques enable biologists to acquire larger, more complex image datasets. Emerging imaging modalities such as multispectral, harmonic, and fluorescence lifetime can generate data with six or more dimensions; however, existing software is not well suited to the visualization or analysis of such data. To address these concerns, we have developed VisBio, an application and toolkit for visualization and analysis of multidimensional, biological image data of any dimensionality. PMID- 15117316 TI - Requirement for the Rac GTPase in Chlamydia trachomatis invasion of non phagocytic cells. AB - Chlamydiae are gram-negative obligate intracellular pathogens to which access to an intracellular environment is paramount to their survival and replication. To this end, chlamydiae have evolved extremely efficient means of invading nonphagocytic cells. To elucidate the host cell machinery utilized by Chlamydia trachomatis in invasion, we examined the roles of the Rho GTPase family members in the internalization of chlamydial elementary bodies. Upon binding of elementary bodies on the cell surface, actin is rapidly recruited to the sites of internalization. Members of the Rho GTPase family are frequently involved in localized recruitment of actin. Clostridial Toxin B, which is a known enzymatic inhibitor of Rac, Cdc42 and Rho GTPases, significantly reduced chlamydial invasion of HeLa cells. Expression of dominant negative constructs in HeLa cells revealed that chlamydial uptake was dependent on Rac, but not on Cdc42 or RhoA. Rac but not Cdc42 was found to be activated by chlamydial attachment. The effect of dominant negative Rac expression on chlamydial uptake is manifested through the inhibition of actin recruitment to the sites of chlamydial entry. Studies utilizing Green Fluorescent Protein fusion constructs of Rac, Cdc42 and RhoA, showed Rac to be the sole member of the Rho GTPase family recruited to the site of chlamydial entry. PMID- 15117317 TI - Misfolding of the prion protein at the plasma membrane induces endocytosis, intracellular retention and degradation. AB - Suramin induces misfolding of the cellular prion protein (PrP(C)) and interferes with the propagation of infectious scrapie prions. A mechanistic analysis of this effect revealed that suramin-induced misfolding occurs at the plasma membrane and is dependent on the proximal region of the C-terminal domain (aa 90-158) of PrP(C). The conformational transition induces rapid internalization, mediated by the unstructured N-terminal domain, and subsequent intracellular degradation of PrP(C). As a consequence, PrP Delta N adopts a misfolded conformation at the plasma membrane; however, internalization is significantly delayed. We also found that misfolding and intracellular retention of PrP(C) can be induced by copper and that, moreover, copper interferes with the propagation of the pathogenic prion protein (PrP(Sc)) in scrapie-infected N2a cells. Our study revealed a quality control pathway for aberrant PrP conformers present at the plasma membrane and identified distinct PrP domains involved. PMID- 15117318 TI - Insights into the phosphoregulation of beta-secretase sorting signal by the VHS domain of GGA1. AB - BACE (beta-site amyloid precursor protein cleaving enzyme, beta-secretase) is a type-I membrane protein which functions as an aspartic protease in the production of beta-amyloid peptide, a causative agent of Alzheimer's disease. Its cytoplasmic tail has a characteristic acidic-cluster dileucine motif recognized by the VHS domain of adaptor proteins, GGAs (Golgi-localizing, gamma-adaptin ear homology domain, ARF-interacting). Here we show that BACE is colocalized with GGAs in the trans-Golgi network and peripheral structures, and phosphorylation of a serine residue in the cytoplasmic tail enhances interaction with the VHS domain of GGA1 by about threefold. The X-ray crystal structure of the complex between the GGA1-VHS domain and the BACE C-terminal peptide illustrates a similar recognition mechanism as mannose 6-phosphate receptors except that a glutamine residue closes in to fill the gap created by the shorter BACE peptide. The serine and lysine of the BACE peptide point their side chains towards the solvent. However, phosphorylation of the serine affects the lysine side chain and the peptide backbone, resulting in one additional hydrogen bond and a stronger electrostatic interaction with the VHS domain, hence the reversible increase in affinity. PMID- 15117319 TI - Sorting of H,K-ATPase beta-subunit in MDCK and LLC-PK cells is independent of mu 1B adaptin expression. AB - The cytoplasmic tail of the H,K-ATPase beta-subunit contains a putative tyrosine based motif that directs the beta-subunit's basolateral sorting when it is expressed in Madin-Darby Canine Kidney (MDCK) cells. When expressed in LLC-PK(1) cells, however, the beta-subunit is localized to the apical membrane. Several proteins that contain tyrosine-based motifs, including the low-density lipoprotein and transferrin receptors, show a similar sorting 'defect' when expressed in LLC-PK(1) cells. For low-density lipoprotein and transferrin receptors, this behavior is due to the differential expression of the mu 1B subunit of the AP-1B clathrin adaptor complex. mu 1B is expressed by MDCK cells, but not LLC-PK(1) cells, and transfection of mu 1B into LLC-PK(1) cells restores basolateral localization of low-density lipoprotein and transferrin receptors. For the beta-subunit, however, mu B expression in LLC-PK(1) cells does not induce its basolateral expression. We found that the beta-subunit interacts with both mu 1B and mu 1A in vitro and in vivo. The capacity to participate in a mu 1B interaction therefore is not sufficient to program the beta-subunit's basolateral localization in MDCK cells. Our data suggest that the H,K-ATPase beta-subunit's basolateral sorting signal is either masked in certain epithelial cells, or requires an interaction with sorting machinery other than AP-1B for delivery to the basolateral plasma membrane. PMID- 15117321 TI - Prevention of hepatitis C virus infection. AB - In spite of advances made in our understanding of the biology of the hepatitis C virus (HCV), the epidemiology and natural history of HCV infection, and the treatment of chronic hepatitis C, the development and worldwide implementation of a comprehensive prevention and control strategy remains necessary. A World Health Organization informal consultation with the Viral Hepatitis Prevention Board was convened and met in Geneva, Switzerland, 13-14 May 2002, to review epidemiological and public health aspects of HCV infection, and the various prevention and control strategies that are currently in place. Based on the presentations and discussions, a number of specific recommendations were made, which should be considered in conjunction with previously published recommendations. PMID- 15117320 TI - Epoetin alfa treatment for acute anaemia during interferon plus ribavirin combination therapy for chronic hepatitis C. AB - Infection with the hepatitis C virus (HCV) remains chronic in 75% of infected individuals, in whom it can cause liver inflammation and progressive fibrosis leading to cirrhosis in 20% of patients. A sustained viral response (SVR) to HCV therapy, i.e. undetectable plasma HCV RNA 6 months after the end of treatment, leads to permanent eradication of the virus in 98.3% of patients. The current treatment of choice is combination therapy with pegylated interferon alfa (PEG IFN alfa), 2a or 2b, and ribavirin (RBV), which achieves an SVR in 54-56% of patients. In patients with HCV genotype 1, RBV doses of 1000-1200 mg/day are associated with a higher SVR than 800 mg/day (51 vs 40%). However, RBV also causes dose-dependent reversible haemolytic anaemia that, in combination with the myelosuppressive effects of PEG-IFN, results in a mean drop in haemoglobin (Hb) level of 3.7 g/dL within 4 weeks. Conventionally, this acute anaemia has been managed with RBV dose reductions. However, this may result in a decreased SVR rate. Alternatively, this anaemia can be managed with administration of epoetin alfa at 40 000 IU once weekly. In a randomized placebo-controlled trial, treatment with epoetin alfa has been shown to raise Hb levels and maintain RBV doses. Furthermore, the increase in Hb level was associated with improved quality of life. Anaemia in patients treated with interferon plus RBV combination therapy can be managed effectively and safely with once weekly epoetin alfa without sacrificing optimal dosing of RBV. PMID- 15117322 TI - Effects of interferon and ribavirin combination therapy on CD4+ proliferation, lymphocyte activation, and Th1 and Th2 cytokine profiles in chronic hepatitis C. AB - We studied the relationship between immunological markers such as CD4+ proliferation, cytokines profile and lymphocyte activation markers in patients with chronic hepatitis C, having different responses to interferon (IFN) and ribavirin (RBV) treatment. A prospective study of 20 patients was conducted, six had received IFN-alpha-2b alone and 14 IFN in combination with RBV. The proliferative immune responses of peripheral blood mononuclear cells to hepatitis C virus peptides and the lymphocyte activation markers (CD25+, CD38+ and CD69+) were assessed before treatment, at 1 week, and 1, 3 and 6 months of treatment. Cytokines interleukin (IL)-2, IFN-gamma, IL-4 and IL-10 were determined in supernatants before onset of treatment and at 1 and 6 months thereafter. Stimulation indices (SI) were higher in the sustained responders (SR), in comparison with those with no response (NR), before treatment (5.2 +/- 3.7 to 3.3 +/- 1.9, P = 0.028) and also at 6 months (7.8 +/- 1.9 to 4.1 +/- 1.2, P = 0.021). Patients with SR also had high SI to NS3 when compared with those with transitory response or no response (NR) (4.9 +/- 2.5 and 3.3 +/- 1.1, P = 0.033). At 1 month, SR had higher supernatant IL-2 than those with NR (133.8 +/- 119.2 to 56.0 +/- 89.3 pg/mL, P = 0.023) and lower levels of IL-10 (13.8 +/- 10.1 and 167.1 +/- 272.0 pg/mL, P = 0.023) in response to NS3. Combination therapy induced a higher percentage of the lymphocyte activation markers CD69+ and CD38+. In conclusion, we found that SR is associated with higher CD4+ proliferation particularly in response to the NS3 region, promoting a T-helper (Th)1/Th0 profile of cytokines, and that combination therapy induced a higher percentage of lymphocyte activation than therapy with IFN alone. PMID- 15117323 TI - Selective functional deficit in dendritic cell--T cell interaction is a crucial mechanism in chronic hepatitis B virus infection. AB - A defect in specific T cell immunity has long been assumed to be the central mechanism of persistent Hepatitis B virus (HBV) infection. Recent studies on HBV transgenic mice have suggested, however, that functional deficit of dendritic cells (DC) was an underlying cause for the T cell dysfunction. The functions of monocyte-derived DC were determined by studying 75 subjects that included chronic hepatitis B patients with low or high HBV load; antibody to hepatitis B surface antigen (anti-HBs) positive individuals who had recovered completely from previous acute HBV infection; healthy donors who had received hepatitis B vaccination and were anti-HBs positive; and immunologically naive to HBV or the vaccine individual. Impaired interactions between monocyte-derived DC and T cells were shown in chronic HBV infection patients, especially in those with active virus replication. The dysfunctions included: (i) failure of DC to increase human leukocyte antigen (HLA-II), B7 expression and interleukin-12 secretion in responses to hepatitis B surface antigen (HBsAg), (ii) defective induction of T cell proliferative response to HBsAg, (iii) failure to activate T cells to produce cytokines and (iv) deficit in the induction of antigen specific cytotoxic T lymphocytes (CTLs). In vitro treatment of DC with tumour necrosis factor-alpha improved HLA-II and B7 expression, as well as Th cell and CTL responses. It is concluded that defective DC-T cell interactions may account for the specific T cell immune defects in chronic HBV infection. Immunotherapy that aims at restoring DC functions could offer a new opportunity for effectively managing persistent HBV infections. PMID- 15117324 TI - Polymorphisms of NS5B protein relates to early clearance of hepatitis C virus by interferon plus ribavirin: a pilot study. AB - Although randomized trials have shown enhancement of efficacy for combination therapy with interferon (IFN) alpha-2b and ribavirin compared with IFN monotherapy as first-line treatment for chronic hepatitis C, infection with genotype 1b and high viremia are still associated with significantly low response rates compared with non-1 genotypes and low viremia. We analysed amino acid sequences of the viral RNA-dependent RNA polymerase (RdRP) or nonstructural protein 5B (NS5B), responsible for ribavirin misincorporation into RNA products in patients with genotype 1b-related chronic hepatitis C and high viremia, and examined the relationship between such RdRp polymorphisms, and the initial decline in viral load induced by combination therapy with IFN-alpha and ribavirin. Substitution of glutamic acid to lysine at the 124th position (E124K) and of isoleucine to valine at the 85th position (I85V) were found to be closely associated with a potent decline of viral load and viral clearance at 8 weeks of treatment (five of five patients, coincidence rate 100%). In conclusion, our results suggest that the polymorphisms of E124K and I85V identified in NS5B protein are crucial for early viral clearance in patients with genotype 1b and high viremia by combination therapy with IFN and ribavirin, and that detection of amino acid sequence motifs might enable prediction of clinical efficacy. PMID- 15117325 TI - Hepatitis C virus replication kinetics in chimpanzees with self-limited and chronic infections. AB - The availability of molecular beacon-based, real time polymerase chain reaction (PCR) and a semi-automated sample extraction procedure have made it possible for us to retrospectively examine HCV replication kinetics in HCV naive chimpanzees infected during the past 20 years. We compared these in 17 animals that developed chronic infection, and in 21 that developed self-limited infection. No differences were found in infecting dose, or replication kinetics in the acute phase between these two types of infection. An unanticipated finding was the fact that 10 of 17 animals developing chronic infection partially controlled virus replication for 48 +/- 48 weeks after typical acute phase viraemia, and prior to development of chronic infection. Twenty-nine out of 30 (29/30) sera, which were negative by quantitative PCR during the downregulated period, were, however, positive by the more sensitive Genprobe isothermal transcription-mediated amplification (TMA) assay. Thus, downregulation was not complete. Ten animals showing self-limited infection showed complete resolution of viraemia by TMA assay. Quasispecies analysis revealed that in all, except one case, the virus reappearing after downregulation was essentially identical to that of the originally infecting virus. PMID- 15117326 TI - Changes in haemoglobin during interferon alpha-2b plus ribavirin combination therapy for chronic hepatitis C virus infection. AB - Interferon alpha and ribavirin (RBV) combination therapy is associated with decreases in haemoglobin (Hb) concentrations and anaemia. The aim of this analysis was to better characterize the magnitude and frequency of Hb changes and risk factors. This retrospective analysis evaluated treatment-related changes in Hb in 677 patients who participated in either of two interferon alpha-2b plus RBV studies for chronic hepatitis C virus (HCV) infection. Study 1 included 192 interferon alpha-naive patients randomized to receive RBV 1000-1200 mg/day plus interferon alpha-2b 3 million IU daily or three times weekly for 48 weeks. Study 2 included 485 interferon alpha-experienced patients randomized to receive RBV 1000-1200 mg daily plus interferon alpha-2b 3 million IU daily or three times weekly for 4 weeks, followed by three times weekly dosing for 44 weeks. More than 50% of all patients experienced a decrease in Hb > or =30 g/L. Women were 4.4 times as likely as men to experience a Hb level of <100 g/L; however, men were at a 40% higher risk to experience a Hb decline of >30 g/L from baseline. Daily use of interferon alpha-2b did not impact the magnitude of Hb decrease. In this pooled analysis, RBV dose reduction resulted in increases in Hb concentration of approximately 10 g/L. Lower baseline creatinine clearance, higher baseline Hb levels and increased age were independently associated with increased risk of Hb decreases of >27.7%. Lower baseline weight was not associated with increased risk of Hb decrease. Substantial Hb decreases occur frequently with interferon alpha/RBV combination therapy. Sex, the magnitude of the Hb decline and renal function are potentially important factors to consider in patients receiving RBV. Further research is needed to determine the impact on virological response and to develop strategies to manage the medical consequences. PMID- 15117327 TI - Serum immunoglobulins predict the extent of hepatic fibrosis in patients with chronic hepatitis C virus infection. AB - Recently, we documented that immunoglobulins stimulate the proliferative activity of rat hepatic stellate cells in vitro. The aim of the present study was to determine whether there is any association between serum immunoglobulin levels and hepatic fibrosis in patients with chronic hepatitis C virus (HCV) infection. Charts from 116 patients with biochemical, serologic, virologic and histologic evidence of chronic hepatitis C infection and serum immunoglobulin levels (IgA, IgG, IgM and total) were reviewed. The mean (+/-SD) age of the study population was 46 +/- 11 years and 67 (58%) were male. There were significant correlations between serum IgA (r = 0.39, P = 0.00001), IgG (r = 0.49, P = 0.000002) and total (r = 0.51, P = 0.000003) immunoglobulin levels and the stage of hepatic fibrosis. When serum immunoglobulin levels were included into logistic regression analysis with variables known to be associated with advanced disease (male gender, age >40 years at onset of infection, duration of infection beyond 20 years and concurrent alcohol abuse) only IgA, IgG and total immunoglobulin levels (P < 0.05, <0.05 and <0.005, respectively) emerged as independent predictors of hepatic fibrosis. Our data indicate a strong association between serum immunoglobulin levels (IgA, IgG and total) and hepatic fibrosis in patients with HCV infection. This finding supports the need to further investigate whether immunoglobulins independently promote disease progression in patients with chronic HCV infection. PMID- 15117328 TI - Does an 'autoimmune' profile affect the clinical profile of chronic hepatitis C? An Italian multicentre survey. AB - Nonorgan-specific autoantibodies (NOSA) are common in patients with chronic hepatitis C virus infection. It is unclear whether serological markers of autoimmunity segregate in a cohort of cases with more severe liver damage. We assessed the relationship between NOSA and demographic, biochemical and histological features in 502 subjects with anti-HCV positive, HCV-RNA positive, HBsAg negative chronic hepatitis consecutively referred to four Italian liver units. Percutaneous liver biopsy was performed in all subjects. A single pathologist scored the biopsies using histology activity index classification. The overall prevalence of positivity for any NOSA was 36.9%. Antinuclear antibodies, anti-smooth muscle antibodies, and anti-liver/kidney microsomal antibodies were found in 15.7, 27.3 and 2.2% of cases. Multivariate analysis showed that gamma-globulin >2 g/dL was the only independent predictor of the likelihood of NOSA positivity (OR, 2.1; 95% CI, 1.3-3.4). No other clinical (age, gender, ALT, HCV genotype) or histological features (grading and staging score, bile ductular damage) were linked to NOSA. Antiviral therapy in 155 subjects with NOSA did not cause any adverse events related to autoimmunity during and after treatment. The presence of NOSA in patients with chronic HCV hepatitis is not related to specific demographic features and has no impact on the biochemical and histological profile of the liver disease at presentation and the response to antiviral treatment. PMID- 15117329 TI - Immunohistochemistry for the detection of swine hepatitis E virus in the liver. AB - Swine hepatitis E virus (HEV) antigen was detected immunohistochemically in formalin-fixed, paraffin-embedded hepatic tissue from 30 naturally infected pigs. Thirty pigs from 30 different herds were selected on the basis of positive results for reverse transcription-polymerase chain reaction. Positive cells typically exhibited a red reaction product in the cytoplasm without any observable background staining. Swine HEV antigen was consistently detected in liver from all 30 pigs tested. A strong immunohistochemical signal was seen within a variable number of hepatocytes in multifocal lobules. The signal involved the majority of hepatocytes diffusely or was confined to foci of liver cells. Positive immunohistochemical signals were also detected in small and large intestine, lymph node, tonsil, spleen, and kidney. The immunohistochemistry technique developed in this study proved useful for the detection of swine HEV in formalin-fixed, paraffin-embedded tissues taken from naturally infected pigs and may be a valuable tool in studying the pathogenesis of swine HEV infection. PMID- 15117330 TI - Infection with human herpes virus type 8 in an area at high prevalence for hepatitis C virus infection in southern Italy. AB - The Campania Region is a geographical area of southern Italy characterized by high incidence rates of hepatocellular carcinoma and of classic Kaposi's sarcoma. Epidemiological investigations carried out among different population groups in this region have found high prevalence rates of both hepatitis C virus (HCV) and human herpesvirus type 8 (HHV-8). To assess co-infection rates of HCV and HHV-8, we carried out a cross-sectional seroepidemiological study prevalence in Pomigliano d'Arco, a Health District of Campania located 20 km away from Naples. The overall rate of HCV/HHV-8 co-infection was 3.1%, 3.5% among men and 2.7% among women. No difference emerged in the HCV/HHV-8 co-infection rates according to seropositivity for HCV infection, either overall (Mantel Haenszel odds ratio = 1.2, 95% CI: 0.6-2.6) or when the analysis was stratified by gender. These findings support the hypothesis that in Campania common routes of transmission are rarely shared by HCV and HHV-8 infections. Local factors may result in different epidemiological patterns for these two viral infections. However, our findings have important public health implications, especially in Mediterranean countries where HCV and HHV-8 infections are endemic. PMID- 15117331 TI - Single nucleotide polymorphism of the MxA gene promoter influences the response to interferon monotherapy in patients with hepatitis C viral infection. AB - The biological activity of interferon (IFN) is mediated by the induction of intracellular antiviral proteins, such as 2'-5' oligoadenylate synthetase, dsRNA activated protein kinase and MxA protein. Among these, MxA protein is assumed to be the most specific surrogate parameter for IFN action. This study was performed to elucidate whether a single nucleotide polymorphism (SNP) (G/T at nt-88) in the promoter region of the MxA gene influences the response to IFN therapy in patients with chronic hepatitis C virus (HCV) infection. Polymorphisms of the MxA gene in 235 HCV patients were determined by polymerase chain reaction-restriction fragment length polymorphism. The frequency of SNP was compared between sustained responders (n = 78) and nonresponders (n = 157), as determined by biochemical and virological responses to IFN. Multivariate analysis showed that among all patients, HCV genotype, HCV RNA level and the SNP of the MxA gene were independent and significant determinants of the outcome of IFN therapy [odds ratio 3.8 (95% confidence interval 2.0-7.0), P < 0.0001; 0.27 (0.15-0.50), P < 0.0001; 1.8 (1.0-3.4), P = 0.0464, respectively]. Furthermore, among patients with a low viral load (< or =2.0 Meq/mL), MxA-T-positive patients were more likely to show a sustained response compared with MxA-T-negative patients [2.87 (1.3-6.3); 62%vs 36%; P = 0.0075]. Our findings suggested that the SNP of the MxA gene is one of the important host factors that independently influences the response to IFN in patients with chronic HCV infection, especially those with a low viral load. PMID- 15117332 TI - Hepatitis A virus and injecting drug misuse in Aberdeen, Scotland: a case-control study. AB - To describe an epidemiological investigation of an outbreak of hepatitis A virus (HAV) infection among injecting drug users in Aberdeen, Scotland. A case-control study to determine whether transmission was facilitated by poor personal hygiene or through sharing injecting equipment. Cases were more likely to report not washing their hands after using the toilet [odds ratio (OR) = 12.9, 95% confidence interval (CI) = 1.58-105.89] or before preparing food (OR = 4.0, 95% CI = 1.01-15.8), and less likely to have washed their hands prior to preparing drugs (OR = 10.67, 95% CI = 2.14-53.07). Cases were also more likely to report recipient sharing of needles/syringes (OR = 8.27, 95% CI = 1.68-40.57), and to have had injecting contact with someone who was jaundiced (OR = 29.4, 95% CI = 3.18-271.44). The results indicate that the lack of hygiene within the context of individuals gathering to prepare and inject drugs provides ample opportunity for the transmission of HAV. Although the promotion of good hygiene and the avoidance of sharing injecting equipment are important measures in preventing HAV transmission, they are unlikely to effect major behavioural change. Such measures should, therefore, be reinforced by routinely offering HAV vaccine to injectors. PMID- 15117333 TI - Restoration of immunity and reactivation of hepatitis B virus after immunosuppressive therapy in a patient with severe aplastic anaemia. AB - We recently treated a patient with severe aplastic anaemia (SAA) who also had chronic hepatitis B virus (HBV) infection. The HBV serological status at the time of diagnosis of SAA was HBsAg(+) and HBeAg(+). Subsequent analysis of the precore region of HBV DNA showed wild-type. He received anti-thymocyte globulin (ATG) and cyclosporin A (CsA) therapy twice. After each course of ATG infusion and during CsA therapy he developed lymphopenia for 1 and 2.5 months, respectively. His serum alanine aminotransferase (ALT) became normalized during the period of lymphopenia, but the serum HBV viral load increased. When his peripheral lymphocytes count recovered, his ALT became elevated again. Lamivudine was effective to normalize his elevated ALT and suppress viral replication. The phenomenon observed in this case supports the prevailing notion that hepatitis B flare-up in HBV carriers after chemotherapy is caused by an immune-mediated mechanism. Meanwhile, this is the first documented case of SAA who developed HBV reactivation upon recovery of lymphopenia after immunosuppressive therapy. This also highlights the necessity of pre-emptive therapy with lamivudine in SAA/HBsAg(+) patients to receive immunosuppressive therapy with ATG/CsA. PMID- 15117334 TI - Urocortin III, a brain neuropeptide of the corticotropin-releasing hormone family: modulation by stress and attenuation of some anxiety-like behaviours. AB - Following its discovery 20 years ago, corticotropin-releasing hormone (CRH) has been postulated to mediate both hormonal and behavioural responses to stressors. Here, we characterize and describe a behavioural role for the murine gene, UcnIII, which encodes a recently discovered CRH-related neuropeptide, urocortin III. We found that mouse UcnIII is expressed predominantly in regions of the brain known to be involved in stress-related behaviours, and its expression in the hypothalamus increases following restraint. In addition, we found that intracerebroventricular administration of mUcnIII stimulates behaviours that are associated with reduced anxiety, including exploration of an open field and decreased latency to enter the lit compartment of a dark-light chamber, but has no effect on the elevated-plus maze. Finally, we found that mUcnIII does not exert any effects on the hormonal stress response. Based upon our findings, UcnIII may be an endogenous brain neuropeptide that is modulated by stress and stimulates behaviours associated with reduced anxiety. In this capacity, UcnIII may attenuate stress-related behaviours, which may be useful both to help cope with stressful situations as well as to avoid pathology associated with excessive reaction to stressors. PMID- 15117335 TI - Effect of environmental enrichment on stress related systems in rats. AB - The aim of this study was to test whether environmental enrichment alters the status and responsiveness of pituitary-adrenocortical and sympathetic adrenomedullary hormones in rats. Previous studies have shown that rats kept in an enriched environment differ from those kept in standard cages in dendritic branching, synaptogenesis, memory function, emotionality and behaviour. In male Wistar rats kept in an enriched environment for 40 days, we studied basal concentrations of hormones, endocrine responses to 5-HT(1A) challenge and responsiveness and adaptation to repeated handling. Environmental enrichment consisted of large plexiglass cages with 10 rats per cage, which contained variety of objects exchanged three times a week. Rats kept in this enriched environment had higher resting plasma concentrations of corticosterone, larger adrenals and increased corticosterone release to buspirone challenge compared to controls. Lower adrenocorticotropic hormone, corticosterone and adrenaline responses to handling were noticed in rats kept in an enriched environment. Exposure to repeated handling led to a more rapid extinction of corticosterone responses in rats kept in an enriched environment. Thus, environmental enrichment leads to pronounced changes in neuroendocrine regulation, including larger adrenals and increased adrenocortical function, which are so far considered to be indication of chronic stress. PMID- 15117336 TI - Gonadotroph heterogeneity, density and distribution, and gonadotroph-lactotroph associations in the pars distalis of the male equine pituitary gland. AB - The intrapituitary mechanisms regulating gonadotrophin secretion in the horse remain unclear. Here, we examined seasonal and gonadal effects on the gonadotroph and lactotroph populations of male horses with the aim of defining a possible morphological basis for the differential release of gonadotrophins. Pituitaries were collected from: (i) gonadal-intact horses in the breeding season (GBS); (ii) orchidectomized horses in the breeding season (OBS); and (iii) orchidectomized horses in the nonbreeding season (ONBS). Immunohistochemistry was performed using antibodies to the luteinizing hormone (LH) beta subunit, follicle-stimulating hormone (FSH) beta subunit and prolactin. In all groups, gonadotrophs were distributed throughout the pars distalis, with dense populations detected near the borders with the pars tuberalis and pars intermedia. The numbers of LH monohormonal, FSH-monohormonal and bihormonal cells/field were greater in GBS than in OBS and ONBS horses. Similarly, the proportion of gonadotrophs in relation to all pituitary cells was larger in gonadal-intact than orchidectomized horses. In the absence of the gonads, no effects of season were observed on these variables. Interestingly, the relative proportions of gonadotroph subtypes and the LH/FSH gonadotroph ratio were similar among groups. Furthermore, while specific gonadotroph-lactotroph associations were identified in all groups, significant gonadal effects within the breeding season and direct effects of season within orchidectomized horses were detected for the number of lactotrophs. This study reveals a gonadal-independent effect of season on the lactotroph, but not the gonadotroph population of the equine pituitary, and a clear gonadal stimulation of both cell types within the breeding season. We suggest that alterations in intercellular arrangements, rather than changes in the incidence of gonadotroph subtypes, may contribute to the differential release of gonadotrophins and, accordingly, to the intrapituitary control of fertility throughout the male equine annual reproductive cycle. PMID- 15117337 TI - Changes of atrial natriuretic peptide in rat supraoptic neurones during pregnancy. AB - To better understand the role of atrial natriuretic peptide (ANP) in the central regulation of hydro-mineral homeostasis, we analysed its expression in rat hypothalamic neurones during gestation and postpartum. These physiological events are characterized by opposing body fluid regulations. Quantitative in situ hybridization analysis showed that starting from mid-pregnancy, ANP mRNA declined in neurones of the preoptic area, periventricular area, lateral hypothalamus and endorhinal nucleus, and remained low at postpartum. By contrast, magnocellular cells in the supraoptic nucleus (SON) showed four- and 10-fold more ANP mRNA in sections from preterm and postpartum rats, respectively, compared to nonpregnant controls (P < 0.001). Oxytocin mRNA paralleled ANP mRNA expression in the SON, whereas vasopressin mRNA rose in early pregnancy and declined thereafter. High hypothalamic ANP concentration at day 21 of gestation versus nonpregnant rats (3.1 +/- 0.5 versus 1.8 +/- 0.4 ng/mg protein, P < 0.05) suggested that ANP transcript accumulation in the SON is associated with increased utilization of the peptide. The elevation of hypothalamic ANP (two-fold) and ANP receptors by treatment of ovariectomized rats with 17beta-oestradiol (25 micro g/rat, 10 days) was abolished by coadministration of progesterone. Thus, we concluded that elevated oestradiol at term stimulates ANP synthesis and paracrine ANP activation in the hypothalamus. Overall, we provide experimental, anatomical and molecular evidence for ANP regulation in hypothalamic neurones at preterm and after 17beta oestradiol stimulation. Our study supports the concept that ANP expressed in the SON acts as a peptidergic neurotransmitter involved in water and salt regulation during pregnancy and postpartum. PMID- 15117338 TI - Subtypes of alpha1- and alpha2-adrenoceptors mediating noradrenergic modulation of spontaneous inhibitory postsynaptic currents in the hypothalamic paraventricular nucleus. AB - Noradrenergic inputs to the hypothalamic paraventricular nucleus (PVN) play important roles in the regulation of neuroendocrine and autonomic functions. Previous reports show that noradrenaline increases the frequency of spontaneous inhibitory postsynaptic currents (sIPSCs) in a subpopulation of type II neurones, acting via alpha(1)-adrenoceptors (ARs), but reduces this frequency in most type I and another subpopulation of type II neurones, via alpha(2)-ARs on presynaptic GABA neurones. Here, we identified the subtypes of alpha-ARs mediating noradrenaline-induced increases and decreases in the sIPSC frequency of PVN neurones, by using slice patch recordings from PVN neurones. In both type I and II neurones, the noradrenaline-induced decrease in sIPSC frequency was completely blocked by BRL44408 (alpha(2A)-AR antagonist) at 1-3 micro M, which is approximately 1/100 of its equilibrium dissociation constant (pA(2) = 8.0), but not by prazosin (20-100 micro M, alpha(2B/C)-AR antagonist; pA(2) = 7.5). The effect of noradrenaline was mimicked by guanfacine (alpha(2A)-AR agonist) with an EC(50) of 0.1 micro M. In type II neurones, the noradrenaline-induced increase in sIPSC frequency was not blocked by any of the following antagonists: RS17053 (10 micro M, alpha(1A)-AR antagonist), BMY7378 (2 micro M, alpha(1D)-AR antagonist), prazosin (0.1 micro M, alpha(1)-AR antagonist; pA(2) = 10.5), or chloroethylclonidine (10 micro M, alpha(1B/D)-AR antagonist). However, the effect of noradrenaline was blocked by higher concentrations of prazosin (1 micro M) or RS17053 (100 micro M), suggesting the involvement of alpha(1L)-subtype, a low affinity form of alpha(1A)-ARs. Collectively, our results indicate that the alpha(2A)-, or alpha(1L)-ARs on the GABA neurones mediate the noradrenaline induced decreases, or increases in the frequencies of the sIPSCs of PVN neurones, respectively. PMID- 15117339 TI - Diverse effects of stress and additional adrenocorticotropic hormone on digitalis like compounds in normal and nude mice. AB - Digitalis-like compounds (DLC) are steroidal hormones that are synthesized in, and released from, the adrenal gland, whose regulation may be directed by the hypothalamic-pituitary-adrenal (HPA) axis. Increasing evidence points to antitumour properties of these compounds and we hypothesized that the establishment of tumours in athymic nude mice may be facilitated by an abnormal synthesis or secretion of DLC. To explore this hypothesis, DLC concentrations were determined in the plasma, and in adrenal and hypothalamic tissues of nude compared to normal mice under basal conditions, and 30 min after a stress stimulus (i.p. injection of 100 micro l saline) with or without additional adrenocorticotropic hormone (ACTH) 1 micro g/per animal. Simultaneously, plasma corticosterone and serum adrenocorticotropic hormone (ACTH) concentrations were analysed. The basal DLC concentrations were similar in the plasma and the hypothalamus of both strains, whereas the basal adrenal DLC concentration was significantly lower in the nude mice compared to normal mice. The stress stimulus induced in normal mice a significant increase in DLC concentrations in the adrenal gland, the plasma and the hypothalamus. However, in nude mice, it caused an increase only in the adrenal gland and the hypothalamus, whereas the plasma DLC concentration was not affected. In both strains, the administration of ACTH in addition to injection stress did not provoke a further increase in DLC concentrations while inducing a significant increase in plasma corticosterone concentration. Regardless of the applied stimulus, the nude mice expressed significant lower DLC concentrations in the adrenal gland and the plasma compared to normal mice. The low basal adrenal DLC concentration in nude mice and their impaired DLC response towards stress- and ACTH stimulation both support an involvement of DLC in tumorigenesis. PMID- 15117340 TI - Acute intracarotid glucose injection towards the brain induces specific c-fos activation in hypothalamic nuclei: involvement of astrocytes in cerebral glucose sensing in rats. AB - The detection of changes in glucose level constitutes the first step of the control of glucose homeostasis. Glucose sensors are therefore expected to be present in different parts of the body and particularly in the central nervous system. Some studies have already attempted to determine glucose-sensitive cerebral structures either after a glucoprivic stimulus or after prolonged hyperglycaemia. By analogy to beta cells, it was postulated that the glucose sensors in the brain could involve GLUT2, glucokinase and/or ATP-sensitive K(+) channels. Surprisingly, GLUT2 was mainly found in astrocytes. Thus, the aims of the present investigation were to determine, in awake rats: (i) the hypothalamic areas that respond to acute hyperglycaemic condition induced by an intracarotid injection of glucose and (ii) the involvement of astrocytes in glucose-sensing by the use of a glial drug, methionine sulfoximine. Rats were given intracarotid injections of glucose solution to trigger a transient insulin secretion without change in peripheral glycaemia, thus involving only central nervous regulation. Hypothalamic activation was determined by immunodetection of the immediate early gene c-fos protein. Acute glucose injection induces significant activation of arcuate and paraventricular nuclei. This stimulation mainly affects neurones in both nuclei, but also astrocytes in the former as illustrated by double immunohistochemistry (Fos and neuronal nuclei or glial fibrillary acidic protein). After specific impairment of astrocyte metabolism by methionine sulfoximine, cerebral activation disappears in the arcuate nucleus, correlated with the lack of cerebral glucose-induced insulin secretion. Therefore, arcuate and paraventricular hypothalamic nuclei are able to detect acute cerebral hyperglycaemia, leading to a peripheral stimulation of insulin secretion. Arcuate nucleus and more especially astrocytes in this nucleus play a pivotal role in glucose-sensing. PMID- 15117341 TI - Modulation of oestrogen receptor-beta mRNA expression in rat paraventricular and supraoptic nucleus neurones following adrenal steroid manipulation and hyperosmotic stimulation. AB - Magnocellular neurosecretory neurones in the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei express oestrogen receptor beta (ERbeta) but not ERalpha. In the PVN, ERbeta is strongly expressed in the ventromedial parvocellular neurones projecting to the brainstem. We used quantitative in situ hybridization, with (35)S-labelled riboprobes, to study heterologous regulation by manipulating adrenal steroid hormones (72 h after adrenalectomy +/- corticosterone replacement; repeated stress: halothane inhalation, environmental cold, immobilization, each daily for 3 days) in male rats. Adrenalectomy increased ERbeta mRNA expression in the magnocellular PVN and SON, by 2.2 and 2.5 fold, respectively, with no effect in the ventromedial parvocellular PVN neurones. Corticosterone replacement partially prevented the increases in ERbeta mRNA expression in magnocellular PVN and SON neurones. Repeated stress over 72 h had no effect on ERbeta mRNA expression in the magnocellular PVN or SON, but increased expression 1.4-fold in the ventromedial parvocellular PVN neurones. Although consequences of hydromineral balance derangement after adrenalectomy may stimulate magnocellular neurones, strongly stimulating the neurones by giving intact male rats 2% saline to drink for 72 h decreased ERbeta mRNA expression in the magnocellular PVN and SON neurones by approximately 60%, and in the ventromedial parvocellular PVN neurones by 13%. Thus, ERbeta mRNA expression is negatively regulated by basal glucocorticoid secretion in magnocellular PVN and SON neurones, and positively regulated by stress in ventromedial parvocellular PVN neurones. However, ERbeta mRNA expression in magnocellular neurones is negatively linked to hyperosmotic stimulation of the neurones. The 6.25-fold variation in ERbeta mRNA expression in magnocellular neurones from salt-loading to adrenalectomy could alter their sensitivity to oestrogens. Consequently, regulation of oxytocin and vasopressin neurone activity via ERbeta is expected to vary according to their functional state and, in particular, on basal glucocorticoid actions. PMID- 15117342 TI - Relationships between aromatase activity in the brain and gonads and behavioural deficits in homozygous and heterozygous aromatase knockout mice. AB - The present study was carried out to determine whether aromatase knockout (ArKO) mice are completely devoid of aromatase activity in their brain and gonads and to compare aromatase activity in wild-type and ArKO mice, as well as in heterozygous (HET) mice of both sexes that were previously shown to display a variety of reproductive behaviours at levels intermediate between wild-type and ArKO mice. Aromatase activity was extremely low, and undetectable by the tritiated water assay, in homogenates of the preoptic area-hypothalamus of adult wild-type mice, but was induced following a 12-day treatment with testosterone. The induction of aromatase activity by testosterone was significantly larger in males than in females. Even after 12 days exposure to testosterone, no aromatase activity was detected in the brain of ArKO mice of either sex whereas HET mice showed intermediate levels of activity between ArKO and wild-type. Aromatase activity was also undetectable in the ovary of adult ArKO females but was very high in the wild-type ovary and intermediate in the HET ovary. In wild-type mice, a high level of aromatase activity was detected on the day of birth even without pretreatment with testosterone. This neonatal activity was higher in males than in females, but females nevertheless appear to display a substantial level of oestrogen production in their brain. Aromatase activity was undetectable in the brain of newborn ArKO males and females and was intermediate between wild-type and ArKO in HET mice. In conclusion, the present study confirms that ArKO mice are unable to synthesize any oestrogens, thereby validating the ArKO mouse as a valuable tool in the study of the physiological roles of oestradiol. In addition, it demonstrates that the intermediate behaviour of HET mice presumably reflects the effect of gene dosage on aromatase expression and activity, that aromatase activity is sexually differentiated in mice during the neonatal period as well as in adulthood and, finally, that the neonatal female brain produces substantial amounts of oestrogens that could play a significant role in the sexual differentiation of the female brain early in life. PMID- 15117344 TI - Nurse, patient and Mr D--dancing the decision-making polka. PMID- 15117346 TI - Consequences count: against absolutism at the end of life. AB - BACKGROUND: There has been a considerable amount of debate in the nursing literature about euthanasia, and especially the distinctions between acts and omissions, and killing and letting die. These distinctions are required by opponents of euthanasia to justify allowing some cases of passive euthanasia while forbidding all cases of active euthanasia. AIM: This paper adds to the debate by arguing that the position that absolutely forbids euthanasia is theoretically inconsistent. METHODS: The paper first considers the place of moral theory in analysing moral problems, within the framework of the principles of biomedical ethics. It is argued that despite a moral pluralism that operates in many areas, the legal status of euthanasia is based upon an absolute deontological position against deliberate killing, which cannot be overridden by appeals to favourable consequences. In order that certain forms of passive euthanasia can be allowed, this position allows distinctions within three pairs of concepts--acts and omissions, killing and letting die, and ordinary and extraordinary means. A further method of justifying certain actions near the end of life is the doctrine of double effect. These paired concepts and the doctrine of double effect are analysed with special reference to their consequences. CONCLUSION: The application of the doctrine of double effect and the three distinctions relies on consideration of their consequences, allowing in practice what in theory is denied. This is important because it weakens the absolute case against euthanasia, which disallows any direct consequentialist appeal. If consequences count in the application of the doctrine and the distinctions, then they should also count directly prior to their application. This strengthens the argument for active euthanasia in certain cases. PMID- 15117347 TI - Cancer symptom transition periods of children and families. AB - BACKGROUND: Children with cancer are reported to experience many symptoms during the cancer trajectory. However, minimal qualitative research has been conducted that explores children's and families' experiences of symptoms. An understanding of the symptom trajectory, grounded in children's and families' experiences, is essential to providing comprehensive and sensitive care to children with cancer and their families. AIM: This paper reports a study designed to explore and describe the symptom course in childhood cancer as experienced by children and their families. DESIGN: Guided by the philosophy of interpretive interactionism, a longitudinal qualitative study was undertaken. A purposive sample of 39 families of children with cancer who resided in Western Canada participated. The children ranged in age from 4.5 to 18 years and varied in their cancer diagnoses. METHODS: Multiple data collection methods included formal and informal interviewing and participant observation. Data were analysed by the constant comparative method. Development of illness narratives added to an understanding of children's and families' experiences. FINDINGS: A substantive theory entitled 'Children's and Families' Lived Experience of Childhood Cancer Symptoms' emerged from the findings. This depicts the experience of cancer in relation to children's changing symptom trajectory. A core category of the theory, 'passage through the transition periods', shows how changing symptom experiences affected children's and families' ways of being in the world. These were reflected in six transition periods: (1) it is just the flu; (2) it is more than the flu; (3) it hits home; (4) it is nasty; (5) it is not so bad, it is pretty good; and (6) it is 'dragsville'. The changing roles and responsibilities of family members, and how the family existed in the cancer world, varied depending on the transition period through which they were passing. CONCLUSIONS: Transition periods not only reinforce the dynamic nature of the experience of childhood cancer but, more importantly, show how symptoms can greatly affect the quality of children's and families' day-to-day living. Interpreting cancer in the context of the symptom trajectory provides nurses with a new perspective for understanding childhood cancer, and will assist in the development of symptom relief strategies that will help to contain symptoms and improve overall quality of life for children and families. PMID- 15117348 TI - Factors influencing decision to breastfeed. AB - BACKGROUND: The low prevalence and short duration of breastfeeding in international studies have highlighted the need for more investigations into the problems. International studies have identified certain factors that affect breastfeeding intentions. Due to insufficient published studies on the local breastfeeding situation, this study specifically investigated factors influencing the breastfeeding decision among primipara women in Hong Kong. AIM: The aim of this paper is to report a study investigating the personal, social, cultural, facilities and environmental and other factors contributing to women's decisions to breastfeed, and to exploring first-time mothers' knowledge of breastfeeding and its influence on their breastfeeding intentions. METHODS: Three private and 10 public hospitals in Hong Kong participated in the study. Both quantitative and qualitative data were collected through questionnaires and subsequently by in depth interviews with 230 first-time mothers 24-48 hours after delivery. RESULTS: The results indicated that personal, cultural, social, and environmental factors are common influencing factors in the decision to breastfeed. Mother's knowledge and attitudes, followed by husband's support, were identified as important in influencing infant feeding choice. Expected local cultural differences were also identified. DISCUSSION: Acknowledgement of the influencing factors could lead to appropriate management of the promotion of breastfeeding. Further research into husband's role, recognition and support for infant feeding and the issue of 'inadequate breast milk' is necessary for a successful breastfeeding campaign. Some other interesting findings typical of the local culture, such as family communication and living environment relating to breastfeeding, are also discussed. CONCLUSIONS: The study has identified the influencing factors that affect women's decision to breastfeed. Recommendations are made for health care professionals and the community to create a more baby-friendly atmosphere and environment for the promotion of breastfeeding. PMID- 15117349 TI - Comfort on a ward for older people. AB - BACKGROUND: Comfort is often considered to be a central part of nursing, although the value placed on it may have diminished over time. Many views of comfort are expressed in the literature but it still remains a diffuse concept that requires further clarification. Research evidence about older people and comfort is limited, and further work is needed to find out how staff and patients view comfort and how it is achieved in practice. AIMS: This paper reports a study to investigate what comfort means both to older people in hospital and their health care workers. RESEARCH METHODS: Ethnography was the methodology chosen, and data were collected using in-depth interviews with 19 older people and 27 staff members, and 130 hours of participant observation, complemented by additional weekly visits to the study ward. FINDINGS: Three themes were identified: the nature of comfort/discomfort; key determinants of comfort/discomfort; and the underlying factors that influence the achievement of comfort/discomfort. DISCUSSION: In this hospital setting, the focus of nursing on relief of discomfort suggested a tendency to react to problems, rather than proactively to create an environment that facilitated comfort. Staff were aware of ideals of practice but found these difficult to achieve in reality. In this study comfort was not consistently provided, and some ways of working actively promoted discomfort. As comfort is central to nursing and nursing is central to the care of hospitalized older people, it is crucial that practitioners are enabled to fulfil their potential in this area. PMID- 15117350 TI - Nurses' Observation Scale for Inpatient Evaluation: reliability update. AB - BACKGROUND: The Nurses' Observation Scale for Inpatient Evaluation is a 30-item scale designed in 1965 for behavioural and observational rating of psychiatric inpatients. AIM: This paper reports a re-evaluation of the Nurses' Observation Scale for Inpatient Evaluation carried out to confirm that it remains reliable in a modern United Kingdom (UK) setting. METHOD: The scale is a joint-rated one and we tested the degree of agreement between two individual raters and not, as in previous studies, between two pairs of raters. A total of 100 patients were each rated by two nurses, and a least-squares simple regression model was used to describe the average level of agreement between the pairs of ratings. RESULTS: The correlation in total scale scores was 0.76 (F = 136, P < 0.0001). The correlations for Negative and Positive Factors were 0.68 (P < 0.001) and 0.75 (P < 0.001), respectively. CONCLUSION: This study has shown that the Nurses' Observation Scale for Inpatient Evaluation retains satisfactory inter-rater reliability with current clinical populations, and we suggest that it remains a useful tool for everyday clinical practice and a basis for meaningful communication between staff about patient status. PMID- 15117351 TI - The ethics of non-intervention in a study of patients awaiting coronary artery bypass surgery. AB - BACKGROUND: Nurses conducting clinical research frequently encounter ethically challenging situations that require careful analysis if the decisions taken are to be in the best interests of participants, researchers and society. There is a lack of literature which discusses the ethical aspects of the nurse's role in clinical research studies. AIM: The aim of this paper is to discuss the ethical conflicts and decisions taken during a combined qualitative and quantitative study of patients' experiences whilst awaiting coronary bypass surgery. DISCUSSION: Ethical conflict arose because, despite having gained ethics approval for a non-interventional study, the researcher became concerned for the health of some research participants, whose condition was observed to have deteriorated. During the course of the study four of the 70 participants died. As a result, changes to the original research protocol were negotiated and subsequently the researcher intervened in cases where participants' clinical condition had worsened. CONCLUSION: Nurses conducting clinical research studies can face serious ethical dilemmas, particularly if participants' health is at risk. This paper demonstrates the potential for both role conflict and role convergence in nursing research. We contend that since the roles of researcher and clinician are not mutually exclusive, the interface between the two requires further discussion. The paper may to help inform other researchers who struggle with the issue of non-intervention when presented with research participants in need of professional nursing care. PMID- 15117352 TI - Nursing resistance as ethical action: literature review. AB - BACKGROUND: Much has been written about nursing as a predominantly female profession whose members display passivity, submission, obedience and powerlessness. Alternatively, some authors have presented evidence of nurses' capacity to exercise power, revealing the possible relationship between powerlessness and ethical compromise. Thus, empowerment strategies for nurses can yield ethical action. AIM: The aim of this paper is to use analysis of the literature to demonstrate how the actions and responses of nurses to ethical concerns are examples of nurses exercising power. METHOD: Empirical studies published in the nursing literature between 1990 and 2003 have been analysed to illustrate how nurses' actions of resistance can ensure that moral values are realized in practice. Foucauldian notions of power relations and feminist ethics provide the theoretical framework. CONCLUSIONS: Nurses were found to resist in situations where they experienced moral conflicts in relation to the actions of health professionals; however, instances were cited where they did not. Consequently, strategies for nursing education and management are proposed to increase nurses' understanding of the potential acts of resistance that they could employ in situations of moral conflict or concern. PMID- 15117353 TI - Home care needs of patients with long-term conditions: literature review. AB - BACKGROUND: There is a widely felt need to improve the match between long-term patients' care needs and actual use of home care. As this match is not always adequate, it is important to know what factors influence it. AIM: The aim of this paper is to provide insight into long-term patients' need and actual use of home care, and the factors influencing these. METHOD: A literature review was carried out, based on database searches in PubMed, CINAHL and the Nivel online library catalogue. A total of 114 papers were retrieved, but only 13 clearly dealt with use of professional home care (rather than informal home care or residential care) by people with long-term conditions. RESULTS: There is a dearth of publications on factors influencing the match between care need and actual use of professional home care among people with long-term conditions. Most of the 13 publications reviewed concerned determinants of professional home care use, rather than the match between patients' felt needs and the home care delivered. From these studies, a profile of people with long-term conditions who used home care emerged. In general, older, non-white women, with multiple chronic diseases and impairments, and who had recently had inpatient care, tended to make more use of professional home care. CONCLUSION: Future research in this field is recommended, particularly into system- and patient-related characteristics that may be responsible for the mismatch between care need and use. PMID- 15117354 TI - New low back pain in nurses: work activities, work stress and sedentary lifestyle. AB - BACKGROUND: Low back pain is common among nurses. Previous studies have shown that the risk of low back pain increases rapidly with greater amounts of physical work and psychological stress, but is inversely related to leisure activities. However, these previous studies were predominantly retrospective in design and not many took account of three factors simultaneously. AIMS: This 12-month prospective study examined the relationships between work activities, work stress, sedentary lifestyle and new low back pain. METHODS: A total of 144 nurses from six Hong Kong district hospitals completed a face-to-face baseline interview, which was followed-up by a telephone interview. The main study measures were demographic characteristics, work activities, work stress, physical leisure activities and the nature of new low back pain during the 12-month follow up period. Level of work stress, quality of relationships at work, level of enjoyment experienced at work, and work satisfaction were self-reported. RESULTS: Fifty-six (38.9%) nurses reported experiencing new low back pain. Sedentary leisure time activity was not associated with new low back pain. Being comparatively new on a ward (adjusted relative risk 2.90), working in bending postures (adjusted relative risk 2.76) and poor work relationships with colleagues (adjusted relative risk 2.52) were independent predictors of new low back pain. CONCLUSION: The findings of this study suggest that low back pain is a common problem in the population of nurses in Hong Kong. Being comparatively new on a ward, bending frequently during work and having poor work relationships with colleagues are independent predictors of new low back pain. Training for high risk work activities and ergonomic assessment of awkward work postures are essential. Moreover, relaxation and team-building workshops for nurses, especially those who are less experienced in the type of work on their current ward, are recommended. PMID- 15117355 TI - Uptake of guidelines to avoid and report exposure to blood and body fluids. AB - BACKGROUND: Mucocutaneous and percutaneous exposure to blood and body fluids (inoculation injury) are major risk factors for occupational acquisition of bloodborne infection in health care professionals. Compliance with prescribed 'universal precautions' during exposure-prone procedures has been shown to reduce the risk of acquiring bloodborne viral infection. In addition, reporting such exposures facilitates prophylaxis. AIM: The aim of this paper is to report a study to identify strategies to minimize professionals' risks of acquiring bloodborne infections during exposure-prone procedures. METHOD: All surgeons, theatre nurses who scrub for surgery and midwives employed in general operating theatres and delivery suites within one UK National Health Service trust (n = 276) were surveyed by postal questionnaire. Data were analysed using univariate and bivariate techniques in SPSS version 10. Content analysis was undertaken on the one open-ended question. FINDINGS: The response rate was 72.5% (200/276). Only 1.5% (3/200) of respondents adopted universal precautions for all patients irrespective of whether their bloodborne viral status was known. On average, only half the recommended theatre-specific precautions were always adopted (mean 3.725/7, SD = 1.385). Most respondents (63.3%) admitted making judgements related to nationality, lifestyle or sexual orientation when making decisions about protective clothing. Many respondents (74%, 145/196) reported sustaining an inoculation injury in the 10 years prior to the study. However, under-reporting of injuries was common, and 32.4% (47/145) admitted failing to report injuries. Guideline adherence was influenced by profession, but not by time since qualification. CONCLUSIONS: The findings suggest that strategies must be developed to improve compliance with universal precautions and reporting guidelines by all health care professionals. The extent of, and reasons for, non compliance with both local and national guidelines remain relatively unexplored. PMID- 15117356 TI - Response to: Watson's guest editorial, 'Scientific methods are the only credible way forward for nursing research', Journal of Advanced Nursing (2003) 43, pp. 219 220, and subsequent JAN forum pieces, 44, pp. 546-548. PMID- 15117361 TI - Role of oxidative stress and the antioxidant network in cutaneous carcinogenesis. AB - Melanoma and nonmelanoma skin cancers are among the most prevalent cancers in the human population. Solar ultraviolet radiation is considered a major etiological factor but the relationship between dose, timing, and nature of exposure to tumor development is still unclear. Free radicals are generated by normal physiologic processes, including aerobic metabolism and inflammatory response, but may inflict cellular damage when generation is increased and antioxidant defense mechanisms are overwhelmed. Important findings supporting the free radical hypothesis in skin carcinogenesis are: (1) Reactive oxygen species (ROS) are generated in UVA- and UVB-irradiated skin in excessive doses, (2) the natural cutaneous antioxidant defense is impaired upon UV-exposure, (3) free radicals are involved in all steps of carcinogenesis, (4) supplementation with antioxidants can inhibit skin carcinogenesis, and (5) conditions that increase ROS generation enhance photocarcinogenesis. These findings provide a promising rationale for the development of powerful new antioxidant strategies in the prevention and therapy of skin cancer. PMID- 15117362 TI - Neurofibromatosis type 2 with multiple plexiform schwannomas. AB - Multiple plexiform schwannomas (PS) or neurilemmomas are very rare nerve sheath tumors, showing multiple schwannomas of the skin and other soft tissues, brain, or spinal cord with histopathologic characteristic of interwoven Antonio-A type tissues(1) but without other signs of neurofibromatosis type 2 (NF2). Neurofibromatosis type 2 is much less common than neurofibromatosis type 1 (NF1), with a symptomatic prevalence of 1 in 210,000,(2) and, to the authors' knowledge, only six cases of NF2 with multiple plexiform schwannomas have been reported in the literature.(3-5) We report a rare case of multiple cutaneous plexiform schwannomas associated with characteristic features of NF2 including bilateral acoustic neurilemomas and intracranial meningioma. PMID- 15117364 TI - A comparative study of antibody titers of blister fluid and serum in patients with subepidermal immunobullous diseases. AB - BACKGROUND: Subepidermal autoimmune bullous diseases (SABD) comprise several disorders, such as bullous pemphigoid (BP), cicatricial pemphigoid (CP), epidermolysis bullosa acquisita (EBA), herpes gestationis (HG), and linear immunoglobulin A (IgA) dermatosis (LAD), and are characterized by antibody production against the basement membrane structures of the skin and mucosa. Although indirect immunofluorescence (IIF) on serum is a routine test for the detection of basement membrane zone antibodies, there have only been a few studies related to IIF on blister fluid. Aim To perform IIF on blister fluid and to compare the results with those of serum. METHODS: IIF on salt-split skin was performed on the serum and blister fluid of 35 patients with SABD (25 bp, three EBA, three HG, three LAD, and one bullous systemic lupus erythematosus) with conjugated IgG, IgA, and C3. RESULTS: Twenty-eight of the 35 patients showed IIF positive blister fluid with a titer similar or less than that of serum. In 25 patients with BP, the most common disease in this study, 23 cases (92%) had positive IIF on serum, 23 cases (92%) on blister fluid, and 24 cases (96%) on either serum or blister fluid. Immunoreactant titers in BP blister fluid and serum did not show significant differences (P > 0.05). Epidermal binding of immunoreactants was the most prevalent staining pattern of IIF on salt-split skin (92%) in BP. CONCLUSIONS: From the findings of this study, the blister fluid of patients with SABD can be used for IIF. Although IIF sensitivity on blister fluid is no more than that on serum, the performance of this test on blister fluid in addition to serum may reduce the number of false negative results of IIF found using either of these two substrates alone. PMID- 15117363 TI - IgG class antibodies from psoriasis patients recognize the 60-KDa heat-shock protein of Streptococcus pyogenes. AB - BACKGROUND: We have previously found that psoriatic patients have IgG autoantibodies that recognize lesions but not autologous normal skin. The reactivity of the autoantibodies can be adsorbed with streptococcal antigens. METHODS: IgG antibodies were determined by immunoblot and ELISA to streptococcal antigens and by ELISA to the recombinants HSP60Sp, HSP70Sp, HSP60Ec and HSP60Hu, in plaque (PP) and guttate (GP) psoriasis patients, in healthy subjects (HC) and in individuals with streptococcal throat infections and high ASO titers, but without history of dermatological disease (ISp). RESULTS: We found by immunoblot that the IgG response to 71-, 60-, and 14-kDa protein fractions of Streptococcus pyogenes is important in psoriasis. We also found by ELISA that the response to the rHSP60Sp in PP was higher than in all the other three groups studied (P < 0.05) with an odds ratio of 11.11 (CI95% of 4.33-28.49). The PP infected with S. pyogenes had higher titers of the antirHSP60Sp, high ASO, and high PASI. The PP patients did not significantly recognize the HSP60Ec or the HSP60Hu. The GP patients had a higher response to the rHSP60Sp than the healthy controls or ISp patients (P < 0.05) but showed no association with the disease. The response of the ISp patients to the HSP60Sp was similar to the healthy controls. The response to the rHSP70Sp was similar in the PP patients and the healthy controls. CONCLUSION: Results suggest that a high response to the HSP60Sp could be associated with the chronic form of psoriasis. PMID- 15117365 TI - The role of psychological factors in alopecia areata and the impact of the disease on the quality of life. AB - BACKGROUND: The role of psychological factors in the pathogenesis of alopecia areata (AA) has long been the subject of debate. Numerous studies have provided controversial results. AIM: This case-control study was undertaken to determine the significance of stressful life events and other psychological factors in the etiopathogenesis of AA. The impact of the disease on the quality of life was also assessed. METHODS: Fifty-two adult patients (18 females and 34 males) diagnosed with AA and 52 age- and sex-matched individuals selected from hospital staff without any hair loss (control group) were evaluated using the major life events scale, Beck depression and Beck anxiety inventories, and the Short Form-36 health survey (SF-36). In addition, comparison was performed between two patient groups created according to whether or not they linked emotional trauma with their AA attack. RESULTS: There was no statistically significant difference between the patient and control groups with regard to the total scores of stressful major life events, depression, and anxiety. Of the eight subscales on SF-36, vitality and mental health scores were higher in the control group, whereas social functioning scores were higher in the patients. The total number of stressful life events was higher in patients who attributed their disease to a stressful life event than in those who did not. CONCLUSIONS: It appears unlikely that anxiety and depression play a major role in the etiopathogenesis of AA, but stressful life events may act as a trigger in the onset and/or exacerbation of the disease. Furthermore, AA seems to have a partly negative impact on the health related quality of life. PMID- 15117366 TI - Skin eruption associated with Hymenolepis nana infection. PMID- 15117367 TI - Lipoid proteinosis in two brothers with multiple organ involvement from Saudi Arabia. PMID- 15117368 TI - Postinflammatory hyperpigmentation: a common but troubling condition. PMID- 15117369 TI - Congenital triangular alopecia in association with congenital heart diseases, bone and teeth abnormalities, multiple lentigines and cafe-au-lait patches. PMID- 15117370 TI - Lipoid proteinosis: a case report. PMID- 15117371 TI - Kikuchi-Fujimoto disease: immediate remission with ciprofloxacin. PMID- 15117372 TI - Annular lichen planus involving the esophagus: a rare diagnosis. PMID- 15117373 TI - Cutis laxa-like pseudoxanthoma elasticum with ossification. PMID- 15117374 TI - Epidermolysis bullosa acquisita associated with vitiligo, Graves' disease and nephrotic syndrome. PMID- 15117375 TI - Double-blind, randomized, vehicle-controlled clinical trial of once-daily benzoyl peroxide/clindamycin topical gel in the treatment of patients with moderate to severe rosacea. AB - BACKGROUND: Systemic antibiotics such as tetracycline are well accepted as effective in treating the inflammatory papular/pustular phase of rosacea but may be associated with systemic side-effects. Few controlled data on the use of topical antibiotics in rosacea are available. OBJECTIVE: We evaluated the efficacy and tolerability of a fixed combination of 5% benzoyl peroxide and 1% clindamycin in a topical gel for the treatment of rosacea. Methods This was a 12 week, double-blind, vehicle-controlled, randomized, prospective, parallel-group study in 53 patients with moderate to severe rosacea. RESULTS: The mean percentage reduction in papules and pustules from baseline to the end of treatment was 71.3% in the benzoyl peroxide/clindamycin group (n = 26) and 19.3% in the vehicle group (n = 26; P = 0.0056). A significant (P = 0.0141) difference in favor of benzoyl peroxide/clindamycin was evident by the third week of treatment. Severity scores for erythema, papules/pustules, and flushing/blushing decreased more with benzoyl peroxide/clindamycin than with vehicle. Overall rosacea severity, Physician Global Assessment, and Patient's Global Assessment at the end of treatment were all significantly improved with benzoyl peroxide/clindamycin compared with vehicle (P = 0.0101, 0.0026, and 0.0002, respectively). Application site reactions were reported in four patients (14.8%) in the benzoyl peroxide/clindamycin group. CONCLUSION: A once-daily topical application of a combination of 5% benzoyl peroxide and 1% clindamycin is effective and well tolerated in patients with moderate to severe rosacea. PMID- 15117377 TI - Why is psoriasis uncommon in Africans? The influence of dietary factors on the expression of psoriasis. AB - Psoriasis is uncommonly seen in Africans, probably partly due to genetic factors. However, the dietary habits of Africans may provide another explanation, which is explored in this paper. Maize, the staple diet in most parts of Africa, is high in linoleic acid but low in other polyunstaturated fatty acids and riboflavin. Linoleic acid is a precursor of prostaglandin E2 (PGE2) and its high intake, especially in the absence of other polyunsaturated fatty acids (PUFAs) and riboflavin, results in high tissue production of PGE2. PGE2 is known to suppress cellular immunity, resulting in decreased expression of psoriasis. PMID- 15117376 TI - Localized blastomycosis-like pyoderma with good response to cotrimoxazol and cryotherapy. AB - Blastomycosis-like pyoderma is an unusual, exaggerated, vegetative-tissue reaction to a prolonged primary or secondary bacterial infection. It is a rare disease, usually seen in immunocompromized patients. We report a case of localized blastomycosis-like pyoderma responding poorly to classic treatments, but that gave a dramatic response to a combination treatment of cotrimoxazol and cryotherapy. PMID- 15117378 TI - Coexistence of pemphigus vulgaris with systemic lupus erythematosus. PMID- 15117379 TI - Tinea capitis and topical Terbinafine cream. PMID- 15117380 TI - The roots of cosmetic medicine: hair cosmetics in Byzantine times (AD 324-1453). PMID- 15117383 TI - When I'm 64: Health choices. PMID- 15117384 TI - Evaluating decision aids--where next? AB - Decision aids have been developed to help patients become involved in decision making about their individual health care. During the evaluation of a particular decision aid in maternity care--a set of 10 'Informed Choice' leaflets--we considered the lessons learnt for evaluation of decision aids in the future. Decision aids have been tested mainly in explanatory trials and have been found to be effective. We argue that existing decision aids should be subjected to more pragmatic trials to test their effectiveness in the real world. The small amount of evidence on their use in the real world shows that they face challenges, resulting in poor implementation. Therefore, we propose that implementation strategies are developed which take heed of the findings of research on getting evidence into practice, and in particular address structural barriers such as the lack of time available to health professionals. We recommend that these 'decision aid implementation packages' are developed in conjunction with both health professionals and patients, and identify and address potential barriers to both the delivery of patient involvement in decision-making, and the use of decision aids, in the real world. These 'packages' can then be submitted to pragmatic evaluation. PMID- 15117385 TI - A matter of trust--patient's views on decision-making in colorectal cancer. AB - OBJECTIVES: To determine which aspects of the treatment decision process, therapy and outcomes are most important to patients with colorectal cancer (CRC). DESIGN: Cross-sectional survey. PARTICIPANTS: A total of 102 men and 73 women who had completed primary treatment for CRC in two teaching hospitals in Central Sydney, Australia. MAIN OUTCOMES MEASURES: Patient's rating of the importance of the decision-making aspects and outcomes of treatment for CRC. RESULTS: Trust in their surgeon and confidence of specialty training are of paramount importance to CRC patients. Patients also have a strong desire to get on with treatment quickly and rate the risk of disease recurrence and quality of life as being very important in their treatment decisions. Gender, age and whether the patient had undergone adjuvant radiotherapy were all significant predictors of preferred mode of treatment decision-making. Fifty-eight per cent of women preferred a shared decision-making role compared with 36% of men, whilst older patients and those who had undergone adjuvant radiotherapy were significantly more likely to prefer that their surgeon decide upon treatment when compared with younger patients and respondents who have not had radiotherapy. CONCLUSIONS: Regardless of whether a patient prefers an active or more passive role in decision-making, having a surgeon explain treatment options in a clear, unhurried and open manner is vital to how patients feel about their treatment. Whilst acknowledging that individual patients will have different needs for information and preferences for treatment, there are several factors amongst many in the process of decision-making which are considered very important by patients with CRC. A surgeon who adopts a consultation style that is open and informative, that offers patients the chance to participate in the process of decision-making and clearly explain treatment options and outcomes will engender trust with their patient. PMID- 15117386 TI - Agreement between prostate cancer patients and their clinicians about utilities and attribute importance. AB - PURPOSE: To examine the agreement between prostate cancer patients' utilities for selected health states and their rankings of the importance of six attributes of the health states and the clinicians' judgements of what would be in the patients' best interests. METHOD: Patients with newly diagnosed localized prostate cancer individually completed a time trade-off utility assessment shortly after being diagnosed. The health states evaluated were constructed from a multi-attribute utility model that incorporated six aspects of living with the disease and outcomes of treatment. Each patient assessed his current health state and three hypothetical states that might occur in the future, and provided rankings of the importance of the six attributes. The clinicians caring for each patient independently provided their views of what utilities and importance rankings would be in the patient's best interest. RESULTS: The across-participant correlations between patients' and clinicians' utilities were very low and not statistically significant. Across-participant correlations between patient and clinician importance rankings for the six attributes were also low. Across-health state and across-attribute correlations between utilities or importance rankings were highly variable across patient-clinician pairs. CONCLUSION: In the clinical settings studied, there is not a strong relationship between valuations of current and possible future health states by patients with newly diagnosed prostate cancer and their clinicians. Implications of these results for substituted judgement, when clinicians advise their patients or recommend a treatment strategy, are discussed. PMID- 15117387 TI - Patient-focussed decision-making in early-stage prostate cancer: insights from a cognitively based decision aid. AB - PURPOSE: To study the cognitive processes of early-stage prostate cancer patients as they determined which treatment they preferred, using our cognitively based decision aid. METHOD: The aid was a one-to-one interview that included the structured presentation of information, listing exercises in which the patient identified attributes important to his decision, and trade-off exercises to help him weigh and integrate those attributes together. At various points of the interview, patients identified the attributes they felt were important to their decision, rated their treatment options and completed standardized assessments relating to their decision. In addition, patients participated in a follow-up interview at the time they made their actual treatment decision and again 3 months later. RESULTS: Sixty of 70 (86%) of the invited patients participated in the study. Participating patients identified a median of four important attributes (range 1-10); 36 different attributes were identified at some point in the interview by the group. During the interview, 78% of patients changed which attributes they considered important, and 72% changed their treatment ratings. Stability of treatment choice after the interview and lack of regret after the decision were each positively associated with increasing differentiation between treatment options over time. CONCLUSIONS: The decision process appears to be dynamic for the patients with great variability across patients in what is important to the decision. Increasing stability of choice and lack of regret appear to be related positively to increasing difference over time in how attractive the preferred option is over its closest competitor, rather than to the size of the difference at any one point in time. PMID- 15117388 TI - How is patient-centred care understood by the clinical, managerial and lay stakeholders responsible for promoting this agenda? AB - AIMS AND OBJECTIVES: This study explores how the term patient-centred care is understood, particularly by those who are involved in translating the concept from a theoretical idea into a practical application. It examines the ways in which intermediate level stakeholders such as health service managers, educationalists, professional leaders and officers of patient bodies understand and promote patient-centred care among health professionals actually delivering patient care. DESIGN: Qualitative interview study. SETTING AND PARTICIPANTS: Interviewees were drawn from groups and organizations from four categories: health agencies and regulatory bodies, Royal Colleges and other professional bodies, educational institutions, patient and user groups and consumer organizations. MAIN VARIABLES STUDIED: The meanings and understandings of patient centred care, commitment to implementing patient-centred care and barriers and opportunities to implementation. RESULTS AND CONCLUSIONS: Patient-centred care covers a range of activities from patient involvement in individual care to public involvement in health policy decisions. Current Department of Health policy has made patient-centred care a priority, but has not clarified exactly what it means. Thus, health professionals, educationalists, managers and patient representatives have all developed different meanings of patient-centred care to reflect their own particular backgrounds and roles. The individual aspects of patient-centred care have been neglected in policy terms and important research findings have not been incorporated into policies to change the attitudes and behaviours of health professionals. Developing a shared understanding of patient centred care which encompasses all its components is an important role for the new Commission for Patient and Public Involvement. PMID- 15117389 TI - Consulted but not heard: a qualitative study of young people's views of their local health service. AB - OBJECTIVES: The objective of this study was to identify what children and young people in a health district in a large urban area experience as positive--and not so positive--about their local health services, in the light of a growing expectation that users play a more central role in the design and delivery of services. DESIGN: A qualitative study incorporating a range of methods, including interviews, play techniques and a website. SETTING: Schools, nurseries, community groups, in- and outpatient settings in an inner London health authority. PARTICIPANTS: Young people aged between 4 and 19 years, from community (n = 92) and clinical (n = 57) settings. This included 'hard-to-reach' children, including those leaving care, those in touch with the criminal justice system, asylum seekers, and those with learning disabilities. RESULTS: Alongside planning and environment issues, young people particularly emphasized the impact of communication and relationships with staff on their experience of health services. DISCUSSION AND CONCLUSIONS: Using a range of flexible and age appropriate techniques, young people, even those as young as 4 or 5, are able to comment helpfully on their experiences of service provision. What children had to say is revealing but not astonishing to those working in the National Health Service (NHS). Clinicians and managers, to whom our findings were fed back, made this clear. If we have known for so long that the issues raised here are problems, why are we so poor at acting on this knowledge? The authors suggest four explanations. PMID- 15117390 TI - Developing consumer involvement in rural HIV primary care programmes. AB - OBJECTIVES: As part of a broader medical and psychosocial needs assessment in a rural region of northern California, USA, five focus groups were conducted to explore innovative approaches to creating a system of consumer involvement in the delivery of HIV primary care services in the region. DESIGN: A total of five focus groups (n = 30) were conducted with clients from three of five counties in the region with the highest number of HIV patients receiving primary care. SETTING AND PARTICIPANTS: Participants were recruited by their HIV case managers. They were adults living with HIV, who were receiving health care, and who resided in a rural mountain region of northern California. VARIABLES STUDIED: Group discussions explored ideas for new strategies and examined traditional methods of consumer involvement, considering ways they could be adapted for a rural environment. RESULTS: Recommendations for consumer involvement included a multi method approach consisting of traditional written surveys, a formal advisory group, and monthly consumer led social support/informal input groups. Specific challenges discussed included winter weather conditions, transportation barriers, physical limitations, confidentiality concerns, and needs for social support and education. CONCLUSIONS: A multiple-method approach would ensure more comprehensive consumer involvement in the programme planning process. It is also evident that methods for incorporating consumer involvement must be adapted to the specific context and circumstances of a given programme. PMID- 15117392 TI - Building on the best--choice, responsiveness and equity in the NHS. PMID- 15117391 TI - Ensuring quality information for patients: development and preliminary validation of a new instrument to improve the quality of written health care information. AB - BACKGROUND: Despite the recent focus on improving the quality of patient information, there is no rigorous method of assessing quality of written patient information that is applicable to all information types and that prescribes the action that is required following evaluation. OBJECTIVE: The aims of this project were to develop a practical measure of the presentation quality for all types of written health care information and to provide preliminary validity and reliability of the measure in a paediatric setting. METHODS: The Ensuring Quality Information for Patients (EQIP) tool was developed through a process of item generation, testing for concurrent validity, inter-rater reliability and utility. Patient information managers and health care professionals tested EQIP in three annual audits of health care leaflets produced by a children's hospital. RESULTS: The final tool comprised 20 items. Kendall's tau B rank correlation between EQIP and DISCERN was 0.56 (P = 0.001). There was strong agreement between intuitive rating and the EQIP score (Kendall's tau B = 0.78, P = 0.009). Internal consistency using Cronbach's alpha was 0.80. There was good agreement between pairs of raters (mean kappa = 0.60; SD = 0.18) with no differences based on types of leaflets. Audits showed significant improvement in the number of leaflets achieving a higher quality EQIP rating over a 3-year period. CONCLUSIONS: EQIP demonstrated good preliminary validity, reliability and utility when used by patient information managers and healthcare professionals for a wide variety of written health care information. EQIP uniquely identifies actions to be taken as a result of the quality assessment. Use of EQIP improved the quality of written health care information in a children's hospital. Wider evaluation of EQIP with written information for other populations and settings is recommended. PMID- 15117396 TI - The feasibility of neuropsychological endophenotypes in the search for genes associated with bipolar affective disorder. AB - OBJECTIVE: Efforts to identify genetic loci for bipolar disorder (BPD) have thus far proved elusive. The identification of processes mediating between genotype and phenotype (endophenotypes) may help resolve the carrier status of family members in genetic studies of polygenetic disorders with imperfect penetrance, such as BPD. We reviewed the literature to determine if neuropsychological measures could be used as effective endophenotypes to aid molecular genetic studies searching for genes predisposing to BPD. METHODS: Four prerequisites for endophenotypic markers are described, and a critical review of relevant literature was undertaken to determine if neurocognitive measures satisfy these four requirements in BPD. RESULTS: We found evidence that executive functions and declarative memory may be candidate neurocognitive endophenotypes for BPD. However, we cannot exclude other areas of cognition as being affected by BPD susceptibility genes, given the limits of the current knowledge of the neuropsychology of BPD. In particular, the paucity of studies measuring cognition in healthy relatives of BPD patient limits conclusion regarding familial aggregation of particular neurocognitive deficits (i.e. attention). Furthermore, the effects of clinical state and/or medication usage on cognitive functioning in BPD probands should be further explored. CONCLUSIONS: Molecular genetic studies of BPD may benefit from the application of select neuropsychological measures as endophenotypic markers. The use of these markers, once defined, may improve power for detecting genes predisposing to BPD and may help to better define diagnostic criteria. PMID- 15117397 TI - A functional magnetic resonance imaging study of cortical asymmetry in bipolar disorder. AB - OBJECTIVES: Individuals with bipolar disorder (BPD) exhibit motor, perceptual, and cognitive disturbances involving predominantly right hemisphere dysfunction. This asymmetry has been used to advance the hypothesis that the pathogenesis of bipolar disorder may be related to disturbances of the right cerebral hemisphere. We employed functional magnetic resonance imaging to examine hemispheric asymmetries in manic and depressed BPD. A secondary goal of the study was to examine effects of psychotropic medications on blood volume changes in the motor cortices. METHODS: We studied 18 right-handed BPD and 13 right-handed normal healthy comparison subjects. Blood oxygen level dependent (BOLD) responses in the primary motor area (M1) and supplementary motor area (SMA) of both hemispheres were elicited during reaction time (RT) tasks. RESULTS: Healthy subjects activated the SMA in a reciprocal fashion with significantly greater activity in the left SMA for right hand trials and the right SMA for left hand trials. Depressed BPD subjects failed to show this normal reciprocity indicating a failure to suppress unwanted activity in the ipsilateral right SMA, whereas manic BPD subjects failed to suppress unwanted ipsilateral SMA activity in both hemispheres. Manic and depressed BPD subjects exhibited greater activity in the left primary motor area suggesting increased cortical excitability. BPD subjects treated with antipsychotics or mood-stabilizing medications exhibited longer RTs, lower BOLD responses in M1 and SMA, and a loss of normal hemispheric asymmetry in the SMA than untreated subjects. CONCLUSIONS: The presence of a right hemisphere disturbance in BPD is consistent with the hypothesis that the right hemisphere may be dominant in mood regulation. The presence of both left and right hemisphere disturbances in mania may explain the coexisting psychotic and affective symptoms observed in this condition. PMID- 15117398 TI - Abnormal frontal white matter tracts in bipolar disorder: a diffusion tensor imaging study. AB - OBJECTIVES: Prefrontal white matter has been hypothesized to be integral to the pathophysiology of bipolar disorder. Recent morphometric studies however, have not observed changes in white matter in bipolar patients. We hypothesized that changes in prefrontal function in bipolar disorder, widely reported in the literature, may be related to a loss of white matter tract integrity with a resultant dysconnectivity syndrome. In this study we utilized diffusion tensor imaging (DTI) to examine prefrontal white matter in patients with bipolar disorder. METHODS: Nine patients with bipolar disorder and nine healthy controls were recruited. DTI and localizing anatomic data were acquired, and regions of interest (ROIs) identified in the prefrontal white matter at 15, 20, 25, and 30 mm superior to the anterior commissure (AC). Fractional anisotropy (FA) and trace apparent diffusion coefficient (TADC) were compared by ROI between study groups. RESULTS: The FA of ROIs 25 and 30 mm above the AC was significantly reduced in patients with bipolar disorder; FA of all ROIs showed high-medium to large effect sizes. No significant group differences were identified in TADC. CONCLUSIONS: Our findings suggest that a loss of bundle coherence is present in prefrontal white matter. This loss of coherence may contribute to prefrontal cortical pathology in patients with bipolar disorder. PMID- 15117399 TI - Impulsivity: a link between bipolar disorder and substance abuse. AB - BACKGROUND: Substance abuse is present in most patients with bipolar disorder and associated with poor treatment outcome and increased risk of suicide. Increased impulsivity may be a link between bipolar disorder and substance abuse. METHODS: First, we compared impulsivity as a stable trait (Barratt Impulsiveness Scale, BIS) and as state-dependent behavioral laboratory performance (Immediate Memory Delayed Memory task, derived from the Continuous Performance Task) in interepisode bipolar and non-bipolar subjects with and without substance abuse. Secondly, we compared impulsivity in interepisode and manic bipolar subjects with and without substance abuse. RESULTS: The BIS scores were increased in interepisode bipolar disorder and in subjects with histories of substance abuse, and were increased further in interepisode bipolar subjects with substance abuse. Performance impulsivity was increased in subjects with substance abuse, regardless of whether they had bipolar disorder. Among subjects with bipolar disorder, after correction for age, BIS scores were increased in those with substance abuse. Performance impulsivity was increased in manic compared with interepisode subjects, regardless of substance abuse history, and was increased in interepisode subjects with substance abuse similarly to manic subjects without substance abuse. These differences could not be accounted for by age, gender, or course of illness. CONCLUSIONS: Trait impulsivity is increased additively in bipolar disorder and substance abuse. Performance impulsivity is increased in interepisode bipolar disorder only if a history of substance abuse is present. This increased predisposition to impulsivity when not manic may contribute to the decrement in treatment outcome and compliance, and increased risk for suicide and aggression, in bipolar disorder with substance abuse. PMID- 15117400 TI - Quetiapine with lithium or divalproex for the treatment of bipolar mania: a randomized, double-blind, placebo-controlled study. AB - OBJECTIVE: Evaluate the efficacy and tolerability of quetiapine (QTP) combined with lithium (Li) or divalproex (DVP) in the treatment of acute mania. METHODS: Patients were randomized to 21 days of double-blind treatment with QTP plus Li/DVP, or placebo (PBO) plus Li/DVP. QTP was rapidly dosed up to a maximum of 800 mg/day; Li was dosed to 0.7-1.0 mEq/L; or DVP to 50-100 microg/mL. RESULTS: Fifty-six of 91 (61.5%) individuals in the QTP + Li/DVP group compared with 49 of 100 (49%) taking PBO + Li/DVP completed the study. A significantly greater mean reduction in total Young Mania Rating Scale (YMRS) score was observed at end point in patients receiving QTP + Li/DVP compared with those in the PBO + Li/DVP group (-13.76 versus -9.93; p = 0.021). The response rate (> or =50% YMRS improvement) was significantly higher in the QTP + Li/DVP group than in PBO + Li/DVP-treated patients (54.3% versus 32.6%; p = 0.005), as was the proportion of patients achieving clinical remission (YMRS < 12) (45.7% versus 25.8%; p = 0.007). Patients receiving QTP + Li/DVP also had a significantly greater improvement in Clinical Global Impressions-Bipolar (CGI-BP) Severity of Illness scores (-1.38 versus -0.78; p = 0.001). The mean last-week dose of QTP was 584 mg/day in patients meeting response criteria. Common adverse events (at least 10% and twice the rate of Li/DVP) in the QTP + Li/DVP group included somnolence, dry mouth, asthenia, and postural hypotension. CONCLUSIONS: Quetiapine combined with either Li or DVP has superior efficacy compared with Li or DVP monotherapy for treating patients with bipolar mania. Combination therapy was well-tolerated and most adverse events were mild, withdrawal because of adverse events being only 5% compared with 6% on Li or DVP monotherapy. PMID- 15117401 TI - Cognitive impairment in euthymic bipolar patients: implications for clinical and functional outcome. AB - OBJECTIVE: Cognitive impairment in bipolar disorder may be a stable characteristic of the illness, although discrepancies have emerged with regard to what dysfunctions remain during remission periods. The aim of this study was to ascertain whether euthymic bipolar patients would show impairment in verbal learning and memory and in executive functions compared with healthy controls. Secondly, to establish if there was a relationship between clinical data and neuropsychological performance. METHODS: Forty euthymic bipolar patients were compared with 30 healthy controls through a battery of neuropsychological tests assessing estimated premorbid IQ, attention, verbal learning and memory, and frontal executive functioning. The effect of subsyndromal symptomatology was controlled. RESULTS: Remitted bipolar patients performed worse than controls in several measures of memory and executive function, after controlling for the effect of subclinical symptomatology, age and premorbid IQ. Verbal memory impairment was related to global assessment of function scores, as well as to a longer duration of illness, a higher number of manic episodes, and prior psychotic symptoms. CONCLUSIONS: Results provide evidence of neuropsychological impairment in euthymic bipolar patients, after controlling for the effect of subsyndromal depressive symptoms, suggesting verbal memory and executive dysfunctions. Cognitive impairment seems to be related to a worse clinical course and poor functional outcome. PMID- 15117402 TI - Episodic memory impairment in bipolar disorder and obsessive-compulsive disorder: the role of memory strategies. AB - BACKGROUND: There is evidence that individuals with bipolar disorder exhibit neuropsychological impairments not only during mood episodes but also when they are euthymic. One of the most consistently reported cognitive problems in euthymic individuals with bipolar disorder is an impairment in verbal episodic memory. Verbal learning and memory depend on individuals' ability to organize verbal information appropriately during learning. The purpose of the present study was (i) to determine whether episodic memory impairment in euthymic individuals with bipolar disorder is mediated by impairments in organization of verbal information during learning and (ii) to compare the characteristics of memory impairment in bipolar disorder with that previously found in obsessive compulsive disorder (OCD). METHODS: Study participants were 30 individuals with DSM-IV bipolar I disorder (BP-I), 30 individuals with DSM-IV OCD and 30 normal control participants matched for age, gender and education. Participants completed the California Verbal Learning Test (CVLT), a well-established measure of verbal learning and memory that enables assessment of verbal organization strategies during learning. RESULTS: Compared with control subjects, both BP-I and OCD participants showed impaired performance in long-delayed free recall and verbal organization strategies during learning. BP-I participants showed greater long-delay free recall difficulties but not greater verbal organization difficulties during learning than OCD participants. For OCD participants, the long-delay recall impairment was mediated by difficulties using verbal organizational strategies during learning. In contrast, the group difference in long-delayed free recall between BP-I and control participants remained significant even when semantic clustering was introduced as a mediator. This indicated that BP-I participants' long-delayed free recall difficulties were mediated to a lesser extent by difficulties using verbal organizational strategies than for OCD participants. CONCLUSIONS: Verbal episodic memory problems in individuals with bipolar I disorder and OCD are mediated to different degrees by difficulties using semantic clustering encoding strategies compared with control participants. PMID- 15117403 TI - Bipolar-panic disorder comorbidity within bipolar disorder families: a study of siblings. AB - OBJECTIVES: Although anxiety disorders often co-occur with bipolar disorder in clinical settings, relatively few studies of bipolar disorder have looked specifically at panic comorbidity. This report examines lifetime panic comorbidity within a sample of families with a history of bipolar disorder. METHODS: One hundred and nine probands with bipolar disorder and their 226 siblings were interviewed as part of a family-genetic study. Logistic regression was used to model bipolar disorder as a predictor of comorbid panic in those with affective disorder, with age at interview and gender included as covariates. RESULTS: The percentage with panic attacks was low in those without affective disorder (3%) compared with those with unipolar depression (22%) or bipolar disorder (32%). Panic disorder was found only in those with affective disorder (6% for unipolar, 16% for bipolar). When bipolar disorder and unipolar disorder were compared, controlling for age and sex, having bipolar disorder was associated with panic disorder (OR = 3.0, 95% CI = 1.1, 7.8) and any panic symptoms (OR = 2.0, CI = 1.0,3.8) and more weakly with the combination of panic disorder and recurrent attacks (OR = 1.8, CI = 0.9, 3.5). CONCLUSIONS: The absence of panic disorder and the low prevalence of any panic symptoms in those without bipolar or unipolar disorder suggest that panic is associated primarily with affective disorder within families with a history of bipolar disorder. Furthermore, panic disorder and symptoms are more common in bipolar disorder than in unipolar disorder in these families. PMID- 15117404 TI - Menstrually related symptom changes in women with treatment-responsive bipolar disorder. AB - OBJECTIVE: In the present study, we aimed to evaluate menstrually related symptom changes in euthymic women with treatment-responsive bipolar disorder (BD) compared with healthy control subjects and investigate the presence of premenstrual dysphoric disorder (PMDD). METHODS: Thirty-four euthymic women with treatment-responsive BD on mood-stabilizers (lithium and/or valproate) for at least 6 months and 35 control subjects with no history of medical/mental disorder between ages of 18 and 35 years with regular menstrual cycles were prospectively followed up for at least two consecutive menstrual cycles using the Daily Record of Severity of Problems-Short Form (DRSP). Each subject was administered the retrospective self-report questionnaire, Premenstrual Assessment Form (PAF), in the first postmenstrual phase of the menstrual cycle. Venous blood samples were collected between 19 and 22 days of menstrual cycle to evaluate ovulation by measuring the serum progesterone levels. RESULTS: The differences in mean age, age of onset of menses, cycle length and bleeding length did not appear to be significantly meaningful between groups. In the retrospective assessment of premenstrual symptom changes, controls complained more than women with BD. More controls showed a 30% change in DRSP and in depressive and physical sub-groups than the women with BD. Controls demonstrated a significant increase compared with treatment-responsive BD patients in total, depressive, anxiety and attention sub-group scores of DRSP from the postmenstrual to the premenstrual phase, whereas the scores of vegetative symptoms of controls and women with BD did not differ significantly during one cycle or both. Significant menstrual cycle effect was observed in both groups. CONCLUSION: Within the limitations of the study, the results suggest that ongoing mood-stabilizing treatment may have a prophylactic effect against premenstrual symptom changes in women with treatment-responsive BD. PMID- 15117405 TI - Resistant bipolar disorder precipitated by Behcet's syndrome. AB - OBJECTIVE: To report a case of a patient with Behcet's syndrome who developed treatment resistant bipolar disorder (BD) several years after the onset of Behcet's syndrome. METHODS: A 62-year-old woman suffering from Behcet's syndrome since the age of 38, who developed a typical BD 6 years after the first manifestations of the syndrome was described. RESULTS: Once BD occurred, Behcet's syndrome became milder, while BD deteriorated and evolved into a rapid cycling illness. Lithium and carbamazepine were ineffective in controlling the affective symptoms, while sodium valproate combined with low doses of carbamazepine and olanzapine resulted in sufficient stabilization of her mood state. CONCLUSIONS: Behcet's syndrome may have been the organic substrate for BD in this case. The appearance of BD in the setting of an organic-immune disorder, like Behcet's syndrome, suggests that such disorders may be the neurobiologic substrate or contributor for BD, at least in certain cases. PMID- 15117406 TI - Clinical management of obsessive-compulsive-bipolar comorbidity: a case series. AB - OBJECTIVE: To assess the clinical characteristics and the optimal treatment of obsessive compulsive symptoms in patients with familial background, current or past symptoms consistent with a bipolar spectrum diagnosis. METHODS: Longitudinal clinical observation of seven cases with obsessive-compulsive-bipolar comorbidity. RESULTS: In the cases presented that met the DSM-IV-TR criteria for obsessive compulsive disorder (OCD), the concurrent symptoms and the clinical course pointed to a different principal diagnosis and suggested therapeutic strategies different from those currently recommended in OCD. Clear-cut onset of symptoms, high prevalence of mood disorders in relatives, revaluation of their history, switch into mania induced by antidepressants, and clinical follow-up suggested a hierarchical priority of bipolar diagnosis. CONCLUSIONS: The presence of the aforementioned atypical symptoms should alert the physician about the possibility of bipolar comorbidity in OCD patients. If the suspicion is confirmed, bipolar diagnosis should have priority, at least from a therapeutic point of view. The first choice in treatment should be with mood stabilizers or second generation antipsychotics. Mood stabilization should be achieved as a first objective. PMID- 15117407 TI - Endometrial response to IVF hormonal manipulation: comparative analysis of menopausal, down regulated and natural cycles. AB - BACKGROUND: Uterine luminal epithelial cell response to different hormonal strategies was examined to determine commonality when an endometrium attains a receptive, stimulated, morphological profile that may lead to successful implantation. METHODS: Endometrial biopsies from 3 cohorts of patients were compared. The tissue samples taken from these patients were categorized into 8 different groups according to their baseline and the hormone regime used. RESULTS: Pre-treatment natural cycle tissue was variable in appearance. Downregulation with a GnRH analogue tissue appeared menopausal in character. HRT after downregulation resulted in tissue uniformity. HRT in menopause resulted in a 'lush' epithelial surface. HST in the natural cycle improved the morphology with significant difference in secretion between the two regimes examined. CONCLUSIONS: Down regulation plus HRT standardized surface appearance but tissue response is significantly different from the natural cycle, natural cycle plus HRT or menopause plus HRT. HRT in menopause reinstates tissue to a state similar to a natural cycle but significantly different from a natural cycle plus HST. HST with a natural cycle is similar to tissue from the natural cycle but significant differences reflect the influence of the particular hormones present (at any point) within the cycle. PMID- 15117408 TI - Arterial heparan sulfate is negatively associated with hyperglycemia and atherosclerosis in diabetic monkeys. AB - BACKGROUND: Arterial proteoglycans are implicated in the pathogenesis of atherosclerosis by their ability to trap plasma lipoproteins in the arterial wall and by their influence on cellular migration, adhesion and proliferation. In addition, data have suggested an anti-atherogenic role for heparan sulfate proteoglycans and a pro-atherogenic role for dermatan sulfate proteoglycans. Using a non-human primate model for human diabetes, studies examined diabetes induced changes in arterial proteoglycans that may increase susceptibility to atherosclerosis. METHODS: Control (n = 7) and streptozotocin-induced diabetic (n = 8) cynomolgous monkeys were assessed for hyperglycemia by measurement of plasma glycated hemoglobin (GHb). Thoracic aortas obtained at necropsy, were extracted with 4 M guanidine HCL and proteoglycans were measured as hexuronic acid. Atherosclerosis was measured by enzymatic analysis of extracted tissue cholesterol. Glycosaminoglycan chains of arterial proteoglycans were released with papain, separated by agarose electrophoresis and analysed by scanning densitometry. RESULTS: Tissue cholesterol was positively associated with hexuronic acid content in diabetic arteries (r = .82, p < .025) but not in control arteries. Glycosaminoglycan chain analysis demonstrated that dermatan sulfate was associated with increased tissue cholesterol in both control (r = .8, p < 0.05) and diabetic (r = .8, p < .025) arteries, whereas a negative relationship was observed between heparan sulfate and tissue cholesterol in diabetic arteries only (r = -.7, p < .05). GHb, which was significantly higher in diabetic animals (8.2 +/- 0.9 vs 3.8 +/- 0.2%, p < .0005) was negatively associated with heparan sulfate in diabetic arteries (r = -.7, p < .05). CONCLUSIONS: These data implicate hyperglycemia induced modifications in arterial proteoglycans that may promote atherosclerosis. PMID- 15117409 TI - [HCO3-]-regulated expression and activity of soluble adenylyl cyclase in corneal endothelial and Calu-3 cells. AB - BACKGROUND: Bicarbonate activated Soluble Adenylyl Cyclase (sAC) is a unique cytoplasmic and nuclear signaling mechanism for the generation of cAMP. HCO3- activates sAC in bovine corneal endothelial cells (BCECs), increasing [cAMP] and stimulating PKA, leading to phosphorylation of the cystic fibrosis transmembrane conductance regulator (CFTR) and increased apical Cl- permeability. Here, we examined whether HCO3- may also regulate the expression of sAC and thereby affect the production of cAMP upon activation by HCO3- and the stimulation of CFTR in BCECs. RESULTS: RT-competitive PCR indicated that sAC mRNA expression in BCECs is dependent on [HCO3-] and incubation time in HCO3-. Immunoblots showed that 10 and 40 mM HCO3- increased sAC protein expression by 45% and 87%, respectively, relative to cells cultured in the absence of HCO3-. Furthermore, 40 mM HCO3- up regulated sAC protein expression in Calu-3 cells by 93%. On the other hand, sAC expression in BCECs and Calu-3 cells was unaffected by changes in bath pH or osmolarity. Interestingly, BCECs pre-treated with10 microM adenosine or 10 microM forskolin, which increase cAMP levels, showed decreased sAC mRNA expression by 20% and 30%, respectively. Intracellular cAMP production by sAC paralleled the time and [HCO3-]-dependent expression of sAC. Bicarbonate-induced apical Cl- permeability increased by 78% (P < 0.01) in BCECs cultured in HCO3-. However for cells cultured in the absence of HCO3-, apical Cl- permeability increased by only 10.3% (P > 0.05). CONCLUSION: HCO3- not only directly activates sAC, but also up regulates the expression of sAC. These results suggest that active cellular uptake of HCO3- can contribute to the basal level of cellular cAMP in tissues that express sAC. PMID- 15117410 TI - Sexual responsiveness is condition-dependent in female guppies, but preference functions are not. AB - BACKGROUND: Variation in mate choice behaviour among females within a population may influence the strength and form of sexual selection, yet the basis for any such variation is still poorly understood. Condition-dependence may be an important source of variation in female sexual responsiveness and in the preference functions for male display traits that she expresses when choosing. We manipulated food intake of female guppies (Poecilia reticulata), and examined the effect on several measures of condition and various components of mate choice behaviour. RESULTS: Diet significantly influenced four measures of female condition: standard length, weight, reproductive status and somatic fat reserves. Diet also significantly affected female sexual responsiveness, but not preference functions: females in good and poor condition prefer the same males. CONCLUSIONS: Variation in female condition within populations is therefore unlikely to influence the direction of sexual selection imposed by female choice. It may, however, influence the strength of sexual selection due to its effects on female responsiveness. The relative importance of female choice as a sexually selective force may also covary with female condition, however, because low responsiveness may result in sneak copulations being relatively more important as a determinant of the paternity of offspring. Differences among populations in mean condition may also influence geographic differences in the strength of sexual selection. PMID- 15117411 TI - A gene expression system offering multiple levels of regulation: the Dual Drug Control (DDC) system. AB - BACKGROUND: Whether for cell culture studies of protein function, construction of mouse models to enable in vivo analysis of disease epidemiology, or ultimately gene therapy of human diseases, a critical enabling step is the ability to achieve finely controlled regulation of gene expression. Previous efforts to achieve this goal have explored inducible drug regulation of gene expression, and construction of synthetic promoters based on two-hybrid paradigms, among others. RESULTS: In this report, we describe the combination of dimerizer-regulated two hybrid and tetracycline regulatory elements in an ordered cascade, placing expression of endpoint reporters under the control of two distinct drugs. In this Dual Drug Control (DDC) system, a first plasmid expresses fusion proteins to DBD and AD, which interact only in the presence of a small molecule dimerizer; a second plasmid encodes a cassette transcriptionally responsive to the first DBD, directing expression of the Tet-OFF protein; and a third plasmid encodes a reporter gene transcriptionally responsive to binding by Tet-OFF. We evaluate the dynamic range and specificity of this system in comparison to other available systems. CONCLUSION: This study demonstrates the feasibility of combining two discrete drug-regulated expression systems in a temporally sequential cascade, without loss of dynamic range of signal induction. The efficient layering of control levels allowed by this combination of elements provides the potential for the generation of complex control circuitry that may advance ability to regulate gene expression in vivo. PMID- 15117412 TI - Roles of hyaluronan in bone resorption. AB - BACKGROUND: Hyaluronan, an unsulfated glycosaminoglycan, while being closely linked to osteoclast function several years ago, has received little attention lately. Given recent new knowledge of hyaluronan's possible cell binding abilities, it is important to re-examine the role of this polysaccharide in bone homeostasis. DISCUSSION: Previously published data demonstrating a linkage between induction of hyaluronan synthesis and osteoclast-mediated bone resorption are reviewed. Suggestions are made involving the cell binding ability of hyaluronan and its potential to mediate osteoclast binding to bone surfaces and its potential to serve as a diffusion barrier and participate in the sealing zone required for osteoclast-mediated bone resorption. SUMMARY: This brief article summarizes previous studies linking HA to bone resorption and suggests roles for hyaluronan in the process of bone resorption. PMID- 15117413 TI - Ventilatory drive and the apnea-hypopnea index in six-to-twelve year old children. AB - BACKGROUND: We tested the hypothesis that ventilatory drive in hypoxia and hypercapnia is inversely correlated with the number of hypopneas and obstructive apneas per hour of sleep (obstructive apnea hypopnea index, OAHI) in children. METHODS: Fifty children, 6 to 12 years of age were studied. Participants had an in-home unattended polysomnogram to compute the OAHI. We subsequently estimated ventilatory drive in normoxia, at two levels of isocapnic hypoxia, and at three levels of hyperoxic hypercapnia in each subject. Experiments were done during wakefulness, and the mouth occlusion pressure measured 0.1 seconds after inspiratory onset (P0.1) was measured in all conditions. The slope of the relation between P0.1 and the partial pressure of end-tidal O2 or CO2 (PETO2 and PETCO2) served as the index of hypoxic or hypercapnic ventilatory drive. RESULTS: Hypoxic ventilatory drive correlated inversely with OAHI (r = -0.31, P = 0.041), but the hypercapnic ventilatory drive did not (r = -0.19, P = 0.27). We also found that the resting PETCO2 was significantly and positively correlated with the OAHI, suggesting that high OAHI values were associated with resting CO2 retention. CONCLUSIONS: In awake children the OAHI correlates inversely with the hypoxic ventilatory drive and positively with the resting PETCO2. Whether or not diminished hypoxic drive or resting CO2 retention while awake can explain the severity of sleep-disordered breathing in this population is uncertain, but a reduced hypoxic ventilatory drive and resting CO2 retention are associated with sleep-disordered breathing in 6-12 year old children. PMID- 15117414 TI - Abnormal systemic venous connection possibly associated with a persistent right umbilical vein; a case report. AB - BACKGROUND: Abnormal venous connections involving a persistent right umbilical vein are rare. In a minority of cases the liver is entirely bypassed and the condition is associated with multiple congenital malformations. CASE PRESENTATION: The described case illustrates a systemic venous drainage that was severely abnormal in a newborn girl with a truncus arteriosus type II congenital heart defect. Injection of contrast medium through the umbilical vein catheter revealed a very peculiar venous connection that passed anterio-laterally through the right hemithorax before crossing in an oblique fashion towards the superior vena cava. CONCLUSIONS: This venous drainage may be the result of a persistent right umbilical vein connecting with the superior vena cava. PMID- 15117415 TI - The Leeds Evaluation of Efficacy of Detoxification Study (LEEDS) project: an open label pragmatic randomised control trial comparing the efficacy of differing therapeutic agents for primary care detoxification from either street heroin or methadone [ISRCTN07752728]. AB - BACKGROUND: Heroin is a synthetic opioid with an extensive illicit market leading to large numbers of people becoming addicted. Heroin users often present to community treatment services requesting detoxification and in the UK various agents are used to control symptoms of withdrawal. Dissatisfaction with methadone detoxification 8 has lead to the use of clonidine, lofexidine, buprenorphine and dihydrocodeine; however, there remains limited evaluative research. In Leeds, a city of 700,000 people in the North of England, dihydrocodeine is the detoxification agent of choice. Sublingual buprenorphine, however, is being introduced. The comparative value of these two drugs for helping people successfully and comfortably withdraw from heroin has never been compared in a randomised trial. Additionally, there is a paucity of research evaluating interventions among drug users in the primary care setting. This study seeks to address this by randomising drug users presenting in primary care to receive either dihydrocodeine or buprenorphine. METHODS/DESIGN: The Leeds Evaluation of Efficacy of Detoxification Study (LEEDS) project is a pragmatic randomised trial which will compare the open use of buprenorphine with dihydrocodeine for illicit opiate detoxification, in the UK primary care setting. The LEEDS project will involve consenting adults and will be run in specialist general practice surgeries throughout Leeds. The primary outcome will be the results of a urine opiate screening at the end of the detoxification regimen. Adverse effects and limited data to three and six months will be acquired. PMID- 15117416 TI - The rating reliability calculator. AB - BACKGROUND: Rating scales form an important means of gathering evaluation data. Since important decisions are often based on these evaluations, determining the reliability of rating data can be critical. Most commonly used methods of estimating reliability require a complete set of ratings i.e. every subject being rated must be rated by each judge. Over fifty years ago Ebel described an algorithm for estimating the reliability of ratings based on incomplete data. While his article has been widely cited over the years, software based on the algorithm is not readily available. This paper describes an easy-to-use Web-based utility for estimating the reliability of ratings based on incomplete data using Ebel's algorithm. METHODS: The program is available public use on our server and the source code is freely available under GNU General Public License. The utility is written in PHP, a common open source imbedded scripting language. The rating data can be entered in a convenient format on the user's personal computer that the program will upload to the server for calculating the reliability and other statistics describing the ratings. RESULTS: When the program is run it displays the reliability, number of subject rated, harmonic mean number of judges rating each subject, the mean and standard deviation of the averaged ratings per subject. The program also displays the mean, standard deviation and number of ratings for each subject rated. Additionally the program will estimate the reliability of an average of a number of ratings for each subject via the Spearman-Brown prophecy formula. CONCLUSION: This simple web-based program provides a convenient means of estimating the reliability of rating data without the need to conduct special studies in order to provide complete rating data. I would welcome other researchers revising and enhancing the program. PMID- 15117417 TI - What is the value of social values? The uselessness of assessing health-related quality of life through preference measures. AB - BACKGROUND: The use of preference-based measures in the evaluation of health outcomes has extended considerably over the last decade. Their alleged advantage over other types of general instruments in the evaluation of health related quality of life (HRQOL), supposedly lies in the fact that preference measures incorporate values or utilities that reflects the value of social preferences through health states. The objective of this study was to determine whether the use of social preference weights or utilities makes any real difference when calculating scores for the Euroqol (EQ5-D) questionnaire, a HRQOL preference based measure. METHODS: Responses to the EQ5-D of a sample of 10,972 patients from 10 countries enrolled in an observational study of the treatment of schizophrenia in Europe were used for this purpose. Two different methods of scoring the EQ-5D where compared: 'weighting the items' of the questionnaire through the UK official weight coefficients, and 'non-weighting the items'. Pearson's, Spearman's, and two-way mixed parametric intraclass correlation coefficients were used to estimate the association of the scores obtained in both ways. RESULTS: The association between weighted and unweighted Euroqol scores was extremely high (Pearson's r = 0.91), as was the association between their ranks (Spearman's rho = 0.93). The intraclass correlation coefficient obtained (0.89) also suggested that the concordance between the score distributions was prominent. CONCLUSIONS: A non-weighted approach to score the EQ5-D is enough to explain a high proportion of variance in scores obtained through the use of utilities. The differential contribution of weights based on population preference values is therefore minimal and, in our opinion, negligible. PMID- 15117418 TI - Targeted mutagenesis of the Sap47 gene of Drosophila: flies lacking the synapse associated protein of 47 kDa are viable and fertile. AB - BACKGROUND: Conserved proteins preferentially expressed in synaptic terminals of the nervous system are likely to play a significant role in brain function. We have previously identified and molecularly characterized the Sap47 gene which codes for a novel synapse associated protein of 47 kDa in Drosophila. Sequence comparison identifies homologous proteins in numerous species including C. elegans, fish, mouse and human. First hints as to the function of this novel protein family can be obtained by generating mutants for the Sap47 gene in Drosophila. RESULTS: Attempts to eliminate the Sap47 gene through targeted mutagenesis by homologous recombination were unsuccessful. However, several mutants were generated by transposon remobilization after an appropriate insertion line had become available from the Drosophila P-element screen of the Bellen/Hoskins/Rubin/Spradling labs. Characterization of various deletions in the Sap47 gene due to imprecise excision of the P-element identified three null mutants and three hypomorphic mutants. Null mutants are viable and fertile and show no gross structural or obvious behavioural deficits. For cell-specific over expression and "rescue" of the knock-out flies a transgenic line was generated which expresses the most abundant transcript under the control of the yeast enhancer UAS. In addition, knock-down of the Sap47 gene was achieved by generating 31 transgenic lines expressing Sap47 RNAi constructs, again under UAS control. When driven by a ubiquitously expressed yeast transcription factor (GAL4), Sap47 gene suppression in several of these lines is highly efficient resulting in residual SAP47 protein concentrations in heads as low as 6% of wild type levels. CONCLUSION: The conserved synaptic protein SAP47 of Drosophila is not essential for basic synaptic function. The Sap47 gene region may be refractory to targeted mutagenesis by homologous recombination. RNAi using a construct linking genomic DNA to anti-sense cDNA in our hands is not more effective than using a cDNA-anti-sense cDNA construct. The tools developed in this study will now allow a detailed analysis of the molecular, cellular and systemic function of the SAP47 protein in Drosophila. PMID- 15117419 TI - A new set of BXD recombinant inbred lines from advanced intercross populations in mice. AB - BACKGROUND: Recombinant inbred (RI) strains are an important resource for mapping complex traits in many species. While large RI panels are available for Arabidopsis, maize, C. elegans, and Drosophila, mouse RI panels typically consist of fewer than 30 lines. This is a severe constraint on the power and precision of mapping efforts and greatly hampers analysis of epistatic interactions. RESULTS: In order to address these limitations and to provide the community with a more effective collaborative RI mapping panel we generated new BXD RI strains from two independent advanced intercrosses (AI) between C57BL/6J (B6) and DBA/2J (D2) progenitor strains. Progeny were intercrossed for 9 to 14 generations before initiating inbreeding, which is still ongoing for some strains. Since this AI base population is highly recombinant, the 46 advanced recombinant inbred (ARI) strains incorporate approximately twice as many recombinations as standard RI strains, a fraction of which are inevitably shared by descent. When combined with the existing BXD RI strains, the merged BXD strain set triples the number of previously available unique recombinations and quadruples the total number of recombinations in the BXD background. CONCLUSION: The combined BXD strain set is the largest mouse RI mapping panel. It is a powerful tool for collaborative analysis of quantitative traits and gene function that will be especially useful to study variation in transcriptome and proteome data sets under multiple environments. Additional strains also extend the value of the extensive phenotypic characterization of the previously available strains. A final advantage of expanding the BXD strain set is that both progenitors have been sequenced, and approximately 1.8 million SNPs have been characterized. This provides unprecedented power in screening candidate genes and can reduce the effective length of QTL intervals. It also makes it possible to reverse standard mapping strategies and to explore downstream effects of known sequence variants. PMID- 15117420 TI - Visualising very large phylogenetic trees in three dimensional hyperbolic space. AB - BACKGROUND: Common existing phylogenetic tree visualisation tools are not able to display readable trees with more than a few thousand nodes. These existing methodologies are based in two dimensional space. RESULTS: We introduce the idea of visualising phylogenetic trees in three dimensional hyperbolic space with the Walrus graph visualisation tool and have developed a conversion tool that enables the conversion of standard phylogenetic tree formats to Walrus' format. With Walrus, it becomes possible to visualise and navigate phylogenetic trees with more than 100,000 nodes. CONCLUSION: Walrus enables desktop visualisation of very large phylogenetic trees in 3 dimensional hyperbolic space. This application is potentially useful for visualisation of the tree of life and for functional genomics derivatives, like The Adaptive Evolution Database (TAED). PMID- 15117421 TI - Hybrid clustering for microarray image analysis combining intensity and shape features. AB - BACKGROUND: Image analysis is the first crucial step to obtain reliable results from microarray experiments. First, areas in the image belonging to single spots have to be identified. Then, those target areas have to be partitioned into foreground and background. Finally, two scalar values for the intensities have to be extracted. These goals have been tackled either by spot shape methods or intensity histogram methods, but it would be desirable to have hybrid algorithms which combine the advantages of both approaches. RESULTS: A new robust and adaptive histogram type method is pixel clustering, which has been successfully applied for detecting and quantifying microarray spots. This paper demonstrates how the spot shape can be effectively integrated in this approach. Based on the clustering results, a bivalence mask is constructed. It estimates the expected spot shape and is used to filter the data, improving the results of the cluster algorithm. The quality measure 'stability' is defined and evaluated on a real data set. The improved clustering method is compared with the established Spot software on a data set with replicates. CONCLUSION: The new method presents a successful hybrid microarray image analysis solution. It incorporates both shape and histogram features and is specifically adapted to deal with typical microarray image characteristics. As a consequence of the filtering step pixels are divided into three groups, namely foreground, background and deletions. This allows a separate treatment of artifacts and their elimination from the further analysis. PMID- 15117422 TI - B.E.A.R. GeneInfo: a tool for identifying gene-related biomedical publications through user modifiable queries. AB - BACKGROUND: Once specific genes are identified through high throughput genomics technologies there is a need to sort the final gene list to a manageable size for validation studies. The triaging and sorting of genes often relies on the use of supplemental information related to gene structure, metabolic pathways, and chromosomal location. Yet in disease states where the genes may not have identifiable structural elements, poorly defined metabolic pathways, or limited chromosomal data, flexible systems for obtaining additional data are necessary. In these situations having a tool for searching the biomedical literature using the list of identified genes while simultaneously defining additional search terms would be useful. RESULTS: We have built a tool, BEAR GeneInfo, that allows flexible searches based on the investigators knowledge of the biological process, thus allowing for data mining that is specific to the scientist's strengths and interests. This tool allows a user to upload a series of GenBank accession numbers, Unigene Ids, Locuslink Ids, or gene names. BEAR GeneInfo takes these IDs and identifies the associated gene names, and uses the lists of gene names to query PubMed. The investigator can add additional modifying search terms to the query. The subsequent output provides a list of publications, along with the associated reference hyperlinks, for reviewing the identified articles for relevance and interest. An example of the use of this tool in the study of human prostate cancer cells treated with Selenium is presented. CONCLUSIONS: This tool can be used to further define a list of genes that have been identified through genomic or genetic studies. Through the use of targeted searches with additional search terms the investigator can limit the list to genes that match their specific research interests or needs. The tool is freely available on the web at http://prostategenomics.org1, and the authors will provide scripts and database components if requested mdatta@mcw.edu PMID- 15117424 TI - Pathogenesis of malignant mesothelioma. AB - Malignant mesothelioma (MM) is a very aggressive tumor that is caused by environmental, biologic, and genetic factors. Among these factors, asbestos plays a major role. The link between asbestos and MM has been firmly established through numerous epidemiologic studies conducted during the past 40 years. However, the causal role of chrysotile asbestos compared with crocidolite asbestos in MM, the method of correctly establishing asbestos exposure, the amount of asbestos necessary to cause MM, and the mechanisms of asbestos tumorigenicity are still being debated. Along with asbestos, Simian virus 40 (SV40), a DNA monkey virus, has recently been implicated in the etiology of MM. Simian virus 40 large T antigen (Tag) and small t antigen (tag) are largely responsible for the carcinogenicity of the virus, and it is possible that SV40 and asbestos are cocarcinogens. Finally, a genetic factor identified in 3 villages in Cappadocia, Turkey, where 50% of individuals die of MM, appears to be the cause of a high incidence of the disease. In these villages, genetic predisposition for MM works together with erionite, a nonasbestos fiber found in the stones used in construction of houses. The diagnosis of MM is made histologically and confirmed through electron microscopy and immunohistochemistry. Currently available therapies for MM prolong survival by a few months at most. An SV40 vaccine is being developed for human use and it is hoped that it may reduce the incidence of MM in asbestos workers. PMID- 15117425 TI - Pharmacology and mechanism of action of pemetrexed. AB - Pemetrexed is a novel multitargeted antifolate that inhibits > or = 3 enzymes involved in folate metabolism and purine and pyrimidine synthesis. These enzymes are thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyltransferase. This agent has broad antitumor activity in phase II trials in a wide variety of solid tumors, and is approved in combination with cisplatin for the therapy of malignant mesothelioma. In a recent phase III trial, pemetrexed demonstrated equivalent efficacy to docetaxel, but with significantly less toxicity, in second-line treatment of non-small-cell lung cancer. The most common and serious toxicities of pemetrexed--myelosuppression and mucositis--have been significantly ameliorated by folic acid and vitamin B12 supplementation. More important, vitamin supplementation has not been shown to adversely affect efficacy in some tumor types. Tumors with codeletion of the methylthioadenosine phosphorylase gene, as a consequence of p16 deletions, may be particularly sensitive to pemetrexed. In this review, the biochemistry and mechanism of action of pemetrexed are discussed. PMID- 15117426 TI - Pemetrexed alone and in combination with platinum compounds in the management of malignant mesothelioma. AB - Malignant pleural mesothelioma is an aggressive but rare malignancy with a dismal prognosis. It is traditionally resistant to chemotherapy. Antifolate agents have recently shown promising data in the treatment of this malignancy. Pemetrexed is a multitargeted antifolate inhibitor of thymidylate synthase and other folate dependent enzymes that has emerged as one of the most active agents in this disease. Several phase I/II trials of pemetrexed as a single agent or in combination with a platinum drug have demonstrated considerable activity in mesothelioma. In a recently published phase III randomized study, pemetrexed/cisplatin showed a significant improvement in survival, response rate, and quality of life compared with single-agent cisplatin. In addition, several trials reported that folic acid and vitamin B12 supplementation significantly reduced the toxicity observed with the use of pemetrexed without affecting the efficacy of the drug. PMID- 15117427 TI - Novel combinations using pemetrexed in malignant mesothelioma. AB - Malignant mesothelioma is an uncommon malignancy that is locally invasive and rapidly fatal. The majority of patients with mesothelioma are not candidates for curative surgical resection. Chemotherapy has yielded only modest results in these patients. Pemetrexed is a multitargeted antifolate that is being evaluated in many tumor types. Recent single-agent data and data in combination with cisplatin have suggested that pemetrexed has therapeutic benefits in patients with malignant mesothelioma. This article summarizes the data regarding pemetrexed in the treatment of malignant mesothelioma and the potential novel combinations involving the drug that may be used in the future for the treatment of the disease. PMID- 15117428 TI - Pemetrexed in the treatment of advanced non-small-cell lung cancer: a review of the clinical data. AB - Pemetrexed is a novel multitargeting antimetabolite that has first-line and second-line activity against non-small cell lung cancer (NSCLC). Phase II studies have shown significant efficacy and a favorable toxicity profile of the combination of pemetrexed plus platinum as first-line therapy for NSCLC. Second line activity against NSCLC was demonstrated in a phase III trial comparing single-agent pemetrexed with docetaxel; in that trial, survival was comparable between these agents but side effects were significantly less for patients who received pemetrexed. Pemetrexed is also an active agent against mesothelioma. A phase III trial comparing pemetrexed plus cisplatin with cisplatin alone showed for the first time a regimen that improves survival in this disease and led to FDA approval of pemetrexed in combination with cisplatin for mesothelioma. As a radiosensitizer, pemetrexed has been well-tolerated when given concurrent with chest radiation, and a phase I study is under way assessing its tolerability in combination with carboplatin in this setting. Pemetrexed is clearly a useful agent in the treatment of thoracic malignancies, and is worthy of further study in combination with other drugs having novel mechanisms of action. PMID- 15117429 TI - Second-line chemotherapy for non-small-cell lung cancer: recent data with pemetrexed. AB - Platinum-based chemotherapy offers a modest survival advantage over best supportive care (BSC) in chemotherapy-naive patients with a good performance status and advanced/metastatic non-small-cell lung cancer (NSCLC). Based on 2 landmark studies that reported improved survival times and quality of life when comparing docetaxel with ifosfamide, vinorelbine, or BSC alone, docetaxel at 75 mg/m(2) given once every 3 weeks has been the standard of care as second-line chemotherapy since 2000. Docetaxel given at this dose and schedule resulted in significant hematologic toxicity, with many patients at risk for neutropenic fever. Pemetrexed is a novel multitargeted antifolate agent with single-agent activity in first- and second-line treatment of NSCLC. A phase III study in 571 patients comparing pemetrexed with docetaxel demonstrated clinically equivalent therapeutic outcomes in second-line treatment of patients with recurrent NSCLC; however, patients on the pemetrexed arm had a more favorable hematologic toxicity profile, with fewer episodes of neutropenia, neutropenic fever, and infections and less use of granulocyte colony-stimulating factor support. Based on these data, pemetrexed is a reasonable second-line chemotherapy option for patients with recurrent, advanced NSCLC. PMID- 15117430 TI - Information management and informatics: need for a modern pathology service. AB - Requirements for information technology in pathology now extend well beyond the provision of purely analytical data. With the aim of achieving seamless integration of laboratory data into the total clinical pathway, "informatics"- the art and science of turning data into useful information--is becoming increasingly important in laboratory medicine. Informatics is a powerful tool in pathology--whether in implementing processes for pathology modernization, introducing new diagnostic modalities (e.g. proteomics, genomics), providing timely and evidence-based disease management, or enabling best use of limited and often costly resources. Providing appropriate information to empowered and interested patients--which requires critical assessment of the ever-increasing volume of information available--can also benefit greatly from appropriate use of informatics. General trends in medical informatics are reflected in current priorities for laboratory medicine, including the need for unified electronic records, computerized order entry, data security and recovery, and audit. The increasing demands placed on pathology information systems in the context of wider developmental change in healthcare delivery are explored in this paper. PMID- 15117431 TI - Value of laboratory studies in assessment of dehydration in children. AB - BACKGROUND: Due to the lack of a reliable way of clinically measuring dehydration, laboratory tests are usually used to improve the accuracy of clinical assessment of dehydration in children. The purpose of this study was to compare the relationship between clinical and laboratory parameters in the assessment of dehydration and to evaluate the improvement of those parameters over time. METHODS: We conducted a retrospective study to assess the relationship between clinical assessment of dehydration and laboratory findings. RESULTS: Three hundred children were eligible for the study. Twenty-six per cent of those with mild dehydration had serum urea concentrations greater than 14.3 mmol/L, compared with 38% and 5% of those with moderate or no dehydration, respectively. Urea concentration showed a good specificity, 95%. Creatinine concentrations and mean pH were similar whether or not dehydration was present. Bicarbonate and base excess concentrations decreased with the increasing severity of dehydration and were significantly greater in subjects with moderate dehydration than in those without. The sensitivity (71%) and specificity (74%) of both tests were rather poor. All groups had an abnormal anion gap, which was significantly greater in those with mild or moderate dehydration. CONCLUSION: This study confirms that there is a discrepancy between clinical assessment and laboratory parameters of dehydration. Urea showed good specificity, and anion gap was the most sensitive laboratory parameter for assessment of dehydration. These findings need further validation. PMID- 15117432 TI - Subclinical hypothyroidism: a comparison of strategies to achieve adherence to treatment guidelines. AB - BACKGROUND: Subclinical hypothyroidism is an entity based on the laboratory findings of a raised serum thyrotrophin (TSH) concentration and a normal free thyroxine (FT(4)) concentration. Patients with subclinical hypothyroidism who also have anti-thyroid peroxidase (TPO) antibodies have a higher conversion to overt hypothyroidism than those without, and treatment with thyroxine is recommended. METHOD: We audited anti-TPO assay requests within two NHS Trust hospitals, against consensus standards, to ascertain whether a cascade approach to anti-TPO testing and direct advice leads to more appropriate prescribing of thyroxine in general practice. RESULTS: Our data show that where anti-TPO status was automatically tested for and clear advice for treatment given, >85% of patients were treated according to the standard required by the consensus document, with >90% of those recommended to be commenced on thyroxine actually doing so. In contrast, where anti-TPO was not routinely assessed, treatment was started in 46% of patients, without clear evidence that this was appropriate. CONCLUSION: In order to better advise clinicians and in accordance with the agreed protocol, laboratory-generated cascade testing for anti-TPO antibodies should be an integral part of the investigation of subclinical hypothyroidism, and reports should contain appropriate interpretation and advice. PMID- 15117433 TI - Comparison of total, complexed and free prostate-specific antigens and their ratios in the detection of prostate cancer in a non-screened population. AB - BACKGROUND: The role of prostate-specific antigen (PSA) isoforms in the detection of prostate cancer in a non-screened population in the UK remains to be determined. METHODS: Consecutive patients undergoing diagnostic transrectal ultrasound-guided biopsy of the prostate were studied. Prior to biopsy, a blood sample was obtained and total, complexed and free PSA concentrations measured. RESULTS: Of the 171 patients included in the study, 103 were found to have prostate cancer. There were significant differences in total and complexed PSA concentrations and in the ratio of free-to-total PSA (all P <0.001) between patients with prostate cancer and those with benign disease. Receiver operating characteristics (ROC) curve analysis showed that the corresponding areas under the curves were similar. Restricting the analysis to the 77 patients who had total PSA concentrations between 2 and 10 micro g/L, ROC curve analysis showed that total and complexed PSA concentrations failed to discriminate between benign and malignant disease. In contrast, the areas under the ROC curve were greater for the free-to-total ratio (P = 0.033). CONCLUSION: These results show that in patients with total PSA concentrations between 2 and 10 micro g/L, the free-to total PSA ratio was superior to total PSA concentration in discriminating between patients with benign and malignant disease. PMID- 15117434 TI - Comparability of serum prostate-specific antigen measurement between the Roche Diagnostics Elecsys 2010 and the Abbott Architect i2000. AB - BACKGROUND: Most prostate cancers are characterized by a significant rise in the production of serum prostate-specific antigen (PSA), which is used widely in screening for prostate cancer. Within New Zealand, PSA is measured on automated immunoassay analysers, including models by Abbott Laboratories (Architect i2000) and Roche Diagnostics (Elecsys 2010). These assays produce similar, but not identical, results. OBJECTIVE: To compare the measurement of PSA between the Elecsys 2010 and the Architect i2000 assay. METHODS: We measured PSA concentrations in each of 194 serum samples using both the Roche Elecsys 2010 and the Abbott Architect i2000 and compared the results using various statistical methods. RESULTS: PSA concentrations measured on the Architect i2000 system were less than those on the Elecsys 2010 system, by an average of 11%. CONCLUSION: Because the results from the two assays are different, reference intervals appropriate to the method of PSA measurement should be used. PMID- 15117435 TI - Composition of LDL particle discriminates between hypercholesterolaemic persons with and without symptoms of coronary heart disease. AB - BACKGROUND: It is well known that high total and LDL cholesterol concentrations are not prerequisites for ischemic heart disease. This study aimed to differentiate between patients with coronary heart symptoms and healthy subjects with high cholesterol concentrations, using other potential risk factors such as oxidation state, body iron status and the oxidative state of the LDL particle. METHODS: Healthy persons were divided in low cholesterol (<6 mmol/L) (LC) and high cholesterol (> or =6 mmol/L) (HC) groups. An additional group consisted of patients with symptoms of coronary heart disease (P). The lipid profile, body iron status, plasma antioxidants and LDL oxidation status were measured. RESULTS: No significant differences could be observed between the LC and HC groups or between the lipid profiles, plasma antioxidants and parameters of body iron status of the HC and patient groups. Some parameters measured on isolated LDL differed between the HC and patient groups, such as LDL oxidative potential (134.47 vs 93.70 min) and LDL alpha-tocopherol (21.83 vs 11.73 micro mol/mmol cholesterol). CONCLUSION: The oxidative state of the LDL particle discriminated between patients with symptoms of coronary heart disease and hypercholesterolaemic persons without symptoms of coronary heart disease. PMID- 15117436 TI - Longitudinal changes of insulin-like growth factors and their binding proteins throughout normal pregnancy. AB - BACKGROUND: Insulin-like growth factors (IGFs) are anabolic proteins that are essential regulators of cell division, differentiation and growth. We describe the longitudinal changes in IGF-I, IGF-II and the binding proteins IGFBP-1, -2 and -3 before and during normal pregnancy. METHOD: Serum samples were taken before conception and then at 12, 24 and 36 weeks of gestation in 41 healthy women with uncomplicated pregnancies. We measured IGF-I using an automated chemiluminescent method, IGF-II and IGFBP-2 using in-house radioimmunoassays (RIAs), and IGFBP-1 and IGFBP-3 using commercial enzyme-linked immunosorbent assay (ELISA) and RIA kits, respectively. Because of the potential haemodilution effects during pregnancy, albumin was also measured in all samples. RESULTS: There was a significant fall in IGF-I during the first (36%) and second trimesters (21%) followed by an increase of 25% at 36 weeks. During pregnancy, the mean IGF-II concentrations fell by 12% at 12 weeks, 8% at 24 and 8% at 36 weeks compared with pre-conception values. When IGF-II results were adjusted for the haemodilution of pregnancy, its concentrations increased. During pregnancy, there was a rapid increase in mean IGFBP-1 levels by 17-fold (12 weeks), 24-fold (24 weeks) and 25-fold (36 weeks). IGFBP-2 concentrations fell after conception but started to increase towards term. This increase was more significant when adjusted for haemodilution. In contrast, IGFBP-3 concentrations increased significantly throughout pregnancy. CONCLUSION: Our data on the physiological changes of IGFs and their binding proteins add further evidence of the vital roles of these hormones throughout normal pregnancy. PMID- 15117437 TI - Can the addition of interpretative comments to laboratory reports influence outcome? An example involving patients taking thyroxine. AB - INTRODUCTION: There is little evidence that the addition of interpretative comments to biochemistry reports can influence outcome for patients. Interpretative comments on thyroid function test (TFT) requests were introduced in Hull in August 1999, providing the opportunity to determine whether feedback on hypothyroid patients taking thyroxine could lead to a reduction in the proportion whose thyroxine was inadequately replaced. PATIENTS AND METHODS: The study comprised 15 584 TFT requests, made from 1 August 1999 to 30 August 2002 by general practitioners (GPs), for 8281 patients taking thyroxine. Under replacement of thyroxine, defined as a TSH concentration above the upper reference limit (i.e. 4.7 mU/L), was usually commented on in the biochemical report. RESULTS: In the first, second and third years following introduction of interpretative comments, the proportions of samples with a TSH concentration of >4.7 mU/L were 21.3%, 17.6% and 16.6%, respectively (chi(2)(trend) = 43.1, P <0.0001). The proportion with a TSH concentration of <0.1 mU/L showed a more modest change, from 12.5% in year 1 to 14.0% and 14.8% in years 2 and 3, respectively (chi(2)(trend) = 22.3, P <0.0001). CONCLUSION: This study shows that in the three years following the introduction of interpretative comments there was a 22% reduction in the number of GPs' samples indicating thyroxine under replacement. It seems likely that these data provide evidence that comments can indeed influence the biochemical outcome of patients. PMID- 15117438 TI - Faecal calprotectin: a new marker for Crohn's disease? AB - BACKGROUND: Gastroenterologists are often hampered by the lack of a reliable, non invasive index of bowel inflammation when establishing a differential diagnosis for patients presenting with chronic diarrhoea. Investigations aim to distinguish between inflammatory bowel disease (IBD) (e.g. Crohn's disease, ulcerative colitis) and irritable bowel syndrome (IBS). As an acute phase protein, faecal calprotectin measurement may be useful in this context. METHODS: A new ELISA based assay for calprotectin was evaluated. The ability of calprotectin to distinguish between patients with IBS and Crohn's disease was studied. RESULTS: The assay showed adequate inter- and intra-batch imprecision and was suitable for routine use in the laboratory. Calprotectin concentration was significantly greater in patients with Crohn's disease compared with controls (n = 25, P <0.001) and patients with IBS (n = 25, P <0.001). CONCLUSION: A single calprotectin measurement may aid gastroenterologists in the differential diagnosis of Crohn's disease and IBS. Its use could decrease the number of invasive or radiological investigations undertaken in the latter group of patients. PMID- 15117439 TI - Dimercaptosuccinic acid loading test for assessing mercury burden in healthy individuals. AB - BACKGROUND: Oral chelation tests have been used to try to define mercury toxicity in individuals with dental amalgams, who are suffering from a variety of non specific symptoms. METHODS: Self-reported healthy individuals volunteered to undergo an oral chelation test using dimercaptosuccinic acid (DMSA) at a dose of 30 mg/kg body weight. Urinary mercury : creatinine ratios were measured pre-dose and 3 h post-dose. RESULTS: Urinary mercury : creatinine ratios were similar to levels previously reported in individuals with symptoms that could have been attributed to mercury toxicity. One volunteer suffered a serious reaction to DMSA. CONCLUSION: The oral chelation test using DMSA may lead to misleading diagnostic advice regarding potential mercury toxicity and can be associated with serious side effects. PMID- 15117440 TI - Pneumatic tube system induced haemolysis: assessing sample type susceptibility to haemolysis. AB - BACKGROUND: The pneumatic tube system (PTS) has been implicated in inducing haemolysis. It is not known whether certain sample types are more susceptible to haemolysis than others. We assessed the level of haemolysis in commonly used sample types in the clinical biochemistry department when transported through the PTS. METHOD: Blood was collected in pairs for different sample types and sent to the laboratory via the pneumatic tube or delivered by a porter. Haemolysis indices were measured spectrophotometrically and compared for each pair of sample type. RESULTS: Our results suggest that plain serum samples are more susceptible to haemolysis than the other sample types when sent through our PTS (P <0.0001). Compared with serum with gel samples, plain serum samples are more prone to haemolysis (P <0.001). This suggests that gel may confer some protection against haemolysis. CONCLUSION: Different hospitals will have varying system configurations and use different sample types. We recommend that each hospital investigate their own system to assess whether haemolysis is a recurring problem in any of the sample types transported. PMID- 15117441 TI - Hyperhomocysteinaemia in a young woman presenting with stroke, associated with methylene tetrahydrofolate reductase C677T homozygosity. AB - We present a case of ischaemic stroke in a 23-year-old woman, associated with homozygous methylene tetrahydrofolate reductase (MTHFR)-C677T and hyperhomocysteinaemia. Her other risk factors for stroke were ostium secundum atrial septal defect and use of oral contraceptives. This case illustrates the need to include plasma homocysteine (Hcy) measurement in investigations following stroke. In the presence of hyperhomocysteinaemia, the MTHFR genotype should be determined. If the index case has the polymorphism, then all first-degree relatives should also be investigated by measurement of plasma Hcy and determination of MTHFR genotype. PMID- 15117442 TI - Limitations of 17-hydroxyprogesterone in investigating neonatal hyponatraemia. AB - Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is a possible cause of hyponatraemia in the neonate. Elevated 17-hydroxyprogesterone (17-OHP) is considered diagnostic of the condition, although there have been reports of anomalous high concentrations, up to 110 nmol/L, in premature, sick infants subsequently shown to have normal adrenocortical function. We describe a case of a 6-week-old girl with a chest infection and hyponatraemia whose plasma 17-OHP concentration was 300 nmol/L, well within the range associated with 21 hydroxylase deficiency. However, there was no genital ambiguity and plasma cortisol was also significantly elevated, raising the possibility of generalized adrenal hyperstimulation rather than CAH. The patient was treated with antibiotic and saline infusions but no steroids. CAH was subsequently excluded by normal 17 OHP and cortisol responses to Synacthen stimulation. In sickness, an increased plasma 17-OHP concentration may not be synonymous with 21-hydroxylase deficiency, even when grossly raised. Simultaneous measurement of plasma cortisol could aid interpretation and avoid potential misdiagnosis, especially in male infants. PMID- 15117443 TI - Testicular germ cell tumour presenting as thyrotoxicosis. AB - A case is reported of a patient who presented to his family doctor with a short history of cough with signs and symptoms of thyrotoxicosis. Carbimazole treatment had little effect and his symptoms worsened to include severe shortness of breath. He was investigated further and found to have multiple lung and liver metastases from an unknown primary site. Biopsy and subsequent post-mortem investigations revealed a testicular tumour and a grossly elevated serum human chorionic gonadotrophin (hCG) concentration. The biochemical and clinical thyrotoxicosis is presumed to be due to the thyrotrophic activity of excess hCG secretion, in a situation analogous to that seen in hydatidiform mole or in hyperemesis gravidarum. PMID- 15117445 TI - Non-invasive assessment of hepatic iron stores by MRI. PMID- 15117447 TI - The lack of epidemiological link between the HIV type 1 infections in Hong Kong and Mainland China. AB - Blood samples were collected from 139 newly reported HIV-1 infections in Hong Kong over a 3-year period between 1999 and 2001, representing 22.8% of all reported cases. A majority of the patients were male (85.6%), Chinese (74%), and adult (97.1%) and acquired HIV-1 through sexual transmission (88.2%). The B and CRF01_AE were the major subtypes detected--49.6% and 44.6%, respectively. Over time, the frequency of CRF01_AE subtype increased, the B subtype decreased, and new subtypes of C (4) 2.8%, B' (1) 0.7%, and CRF07_BC (3) 2.2% emerged. The CRF01_AE subtype was commoner in female, Chinese, heterosexuals, and injection drug users whereas B subtype was commoner in male, white, and people with homosexual/bisexual contacts. There was no common source of infection from the analysis except a discernible cluster of Vietnamese injection drug users with the CRF01_AE subtype. The molecular findings did not suggest an epidemiological link between HIV infection in Hong Kong and Mainland China. Hong Kong's longstanding and extensive methadone treatment network may have contributed to the phenomenon. PMID- 15117448 TI - CD8+CD38+ T cells but not HIV type 1 RNA viral load predict CD4+ T cell loss in a predominantly minority female HIV+ adolescent population. AB - The purpose of this study was to evaluate predictors of HIV-1 disease progression in a cohort of predominantly female and minority adolescents who had acquired their HIV-1 infections through sexual risk behaviors. Subjects were identified from the REACH cohort who were not on antiretroviral therapy for at least 1 year and whose baseline CD4(+) T cells were >300 cells/mm(3). Biomedical and demographic characteristics of the subjects at the start of the study period were evaluated as predictors of CD4(+) T cell loss in univariate and multivariate models. Two-thirds of the 99 subjects meeting the selection criteria were female and 87% were black or Hispanic similar to the REACH cohort as a whole. Higher absolute CD8(+) CD38(+) T cell counts at the start of the assessment period were associated with a greater rate of loss of CD4(+) T cells. HIV-1 RNA viral load was among other potential predictors of HIV-1 disease progression that had no association with the rate of CD4(+) T cell loss in this cohort. This study extends the observed association of higher CD8(+) CD38(+) T cells numbers being predictive of HIV-1 disease progression into predominantly female, minority youth. PMID- 15117449 TI - Changes in mitochondrial DNA copy number in blood cells from HIV-infected patients undergoing antiretroviral therapy. AB - Mitochondrial DNA (mtDNA) copy number was measured in peripheral blood mononuclear cells (PBMCs) from 69 individuals using a real-time NASBA quantitative assay. Patients with HIV infection harbored significantly lower mtDNA copy number in PBMC than HIV-negative controls. Besides, subjects on stavudine-containing regimens showed significantly lower median mtDNA amounts than HIV-positive patients receiving other antiretroviral drugs, and this was associated with higher lactate levels. Thus, either HIV infection itself or treatment with stavudine-containing regimens might induce mtDNA depletion and related metabolic disturbances as hyperlactatemia. PMID- 15117450 TI - Prediction of virological response to lopinavir/ritonavir using the genotypic inhibitory quotient. AB - The predictive value of virological response to lopinavir (LPV)/ritonavir (r) was assessed in 126 HIV-infected patients who failed antiretroviral therapy and had begun a rescue intervention based on LPV/r. At 3 months, subjects with < or =6 protease (PRO) resistance mutations showed a higher rate of virological response (HIV-RNA drop > 1 log or to <50 copies/ml) than patients with >6 PRO resistance mutations (77% versus 48%; p = 0.01). On the other hand, virological responders had greater mean LPV plasma trough levels than nonresponders (6.4 versus 3.9 microg/ml; p = 0.02). A positive correlation was found between LPV trough concentration and viral load reductions at 3 months under LPV/r (r = 0.23; p = 0.017). Overall, virological response was seen in 80.8% of patients with LPV trough levels >4.8 microg/ml while in only 52.5% of patients with lower LPV trough concentrations (p = 0.002). In the multivariate analysis, both < or =6 PRO resistance mutations and LPV trough levels >4.8 microg/ml were independent predictors of virological response to salvage therapy with LPV/r. A genotypic inhibitory quotient (GIQ) was estimated for each patient based on the ratio between LPV trough levels and the number of PRO resistance mutations. A positive strong correlation was found between GIQ and viral load reductions (r = 0.42; p = 0.002). Virological response was seen in 78% of patients with a GIQ >0.7 but only in 41.6% of those with lower GIQ (p = 0.004). When LPV trough levels >4.8 microg/ml, PRO resistance mutations < or =6, and GIQ >0.7 were all included in a stepwise multivariate analysis, GIQ remained as the main independent predictor of response to LPV/r. PMID- 15117451 TI - HIV type 1 pol gene diversity and archived nevirapine resistance mutation in pregnant women in Rwanda. AB - This study aimed to find out whether genetic polymorphisms were present in positions potentially affecting susceptibility to antiretrovirals in non-B subtypes from HIV-1-infected patients in Rwanda. Viral pol gene diversity was investigated by direct sequencing in 43 treatment-naive women. In addition, 10 DNA sequences from uncultured peripheral blood mononuclear cells were analyzed 6 weeks after a single dose of nevirapine (prevention of mother-to-child transmission program). Phylogenetic analyses have shown 34 subtype A1, 6 subtype C, and 2 subtype D strains. In addition, an A/C recombinant between the protease (PR) (subtype A1) and the reverse transcriptase (RT) (subtype C) was identified. In the PR coding region, high numbers of polymorphisms were found, including substitutions in secondary PR resistance sites. PR 35D, 36I, and 37N were always present within subtype A as were PR 93L in subtype C strains. PR 10I/V, 20R, 33F, and 77V were found in subtype A whereas PR 36I was highly prevalent in subtype C strains. The A/C recombinant displayed substitutions related to resistance (PR 10, 33, 36 and RT 118). One nevirapine resistance mutation (RT 181Y/C) was found in proviral DNA after 6 weeks. In conclusion, subtypes A and C are predominant in this cohort in Rwanda. Substitutions similar to secondary protease inhibitor resistance mutations are common before treatment whereas major resistance mutation may be archived after a single dose of nevirapine. Accordingly, the hypothesis of a genetic background effect in non-B strains has to be further addressed in programs of introduction of antivirals in Africa. PMID- 15117452 TI - Long HIV type 1 reverse transcripts can accumulate stably within resting CD4+ T cells while short ones are degraded. AB - We utilized quantitative methods to compare the efficiency of reverse transcription and stability of viral DNA within resting and activated T cells. Highly purified resting CD4(+) T cells and activated T cells from healthy donors were spinoculated with HIV-1(YU-2), then cultured in conditions that maintain both the viability and the quiescence of the resting cells. Spreading infection was suppressed, then kinetic PCR was used to relate the rates of synthesis of short (strong-stop, RU5) and long (gag or U3-gag second strand transfer) viral DNA to the mean number of virions initially bound to each type of cell. As shown previously, activated cells support an initial burst of high-level reverse transcription, which is then followed by a approximately 10-fold decay in cDNA levels over 4.5 days. In resting T cells, although the synthesis of late reverse transcripts was initially approximately 1000-fold less efficient than in activated T cells, the number of these cDNAs per bound input virion rose 10-fold as culture was extended to 4.5 days. The number of late reverse transcripts remained constant for 3 days after the addition of efavirinez, reflecting enhanced stability. In contrast, the short strong-step reverse transcripts were mostly degraded. Thus, late HIV-1 reverse transcripts can accumulate stably in resting T cells in the absence of detectable T cell activation. Defining the underlying basis for the stabilization of late reverse transcripts, and their associated nucleoprotein complexes, may be pertinent to the accumulation of reservoirs of latent HIV-1 in patients, and could provide a target for future therapies. PMID- 15117453 TI - Zn2+ binding to cysteine-rich domain of extracellular human immunodeficiency virus type 1 Tat protein is associated with Tat protein-induced apoptosis. AB - The Tat protein has several functional domains, one of which is the cysteine-rich domain that is a highly conserved region in spite of the presence of many subtypes of human immunodeficiency virus type 1 (HIV-1). Although the cysteine rich domain is a potential site for Zn(2+) binding, it is controversial whether Zn(2+) is substantially essential for the structure and activities of the Tat protein. To study the significance of Zn(2+) in the cysteine-rich domain of the Tat protein particularly released to the extracellular space, we raised the monoclonal antibody (MAb) 5A4, which has an attractive property of recognizing the Zn(2+)-binding Tat(20-41) peptide but not the apo-Tat(20-41) peptide. MAb 5A4 inhibited the trans-activation of the HIV long terminal repeat (LTR) in HeLa-CD4 LTR/beta-gal cells induced by treatment with the recombinant Tat protein, indicating that MAb 5A4 can recognize the full-length Tat protein and inhibit its trans-activity. The antibody also inhibited the apoptosis of Jurkat cells induced by treatment with the released native-Tat-protein-containing supernatant from the culture of HIV-1(JRFL)-infected cells. These results suggest that Zn(2+), whose structure is closely associated with not only the trans-activation of HIV-LTR but also the induction of apoptosis, binds to the extracellular native Tat protein. The Zn(2+)-binding cysteine-rich domain therefore can be a molecular target in the development of an anti-Tat vaccine and agents for the control of extracellular-Tat-protein-mediated pathogenesis leading to the progression of acquired immunodeficiency syndrome. PMID- 15117454 TI - Effects of human immunodeficiency virus type 1 infection on CCR5 and CXCR4 coreceptor expression on CD4 T lymphocyte subsets in infants and adolescents. AB - HIV-1 infection alters expression of CCR5 and CXCR4 on CD4 T cells in adults, although an effect by virus on expression of coreceptor genes in pediatric subjects is unknown. We designed an exploratory study to evaluate surface expression of CXCR4 and CCR5 on CD45RA and CD45RO subsets of CD4 T lymphocytes from 17 HIV-1-infected infants and adolescents and 16 healthy age-matched individuals. While age in the absence of HIV-1 infection was unrelated to coreceptor expression, infection affected coreceptor expression differentially in infants and adolescents. Among infected adolescents, CCR5 and CXCR4 expression was significantly increased on CD4 CD45RO T cells, while CXCR4 was diminished in the CD4 CD45RA subset. Although HIV-1 infection in infants was also associated with increased CXCR4 expression on the CD4 CD45RO subset, in contrast to adolescents, infection in infants had no impact on coreceptor expression within the CD45RA CD4 subset. The proportion of CD4 T cells coexpressing CD45RA and CD45RO was increased by infection in both infants and adolescents. The CD45RA CD45RO subset in culture expressed high levels of CD4, CXCR4, and CD69, an early activation marker, and was highly susceptible to HIV-1 infection and replication. Infection of transitional CD4 T cells coexpressing CD45RA and CD45RO could contribute in part to provirus in either CD45RA or CD45RO subsets. Deleterious effects by HIV-1 infection on CD4 T cell homeostasis were greater in infants then adolescents, indicating that adolescence may be an optimal age group for assessing vaccines to prevent or treat HIV-1 infection. PMID- 15117456 TI - Interaction of mannose-binding lectin with HIV type 1 is sufficient for virus opsonization but not neutralization. AB - Mannose-binding lectin (MBL), a microbe-recognition protein in serum, binds to high mannose glycans on HIV-1 gp120 and has been reported to neutralize the cell line-adapted strain HIV(IIIB). Because HIV primary isolates (PI) are generally more resistant to neutralization by antibodies and considering that PI are produced in primary cells that could alter the number of high mannose glycans on HIV relative to cell lines, we assessed the ability to MBL to neutralize HIV PI. MBL at concentrations up to 50 microg/ml mediated relatively little neutralization (<20%) of HIV PI infection of peripheral blood mononuclear cells (PBMCs). MBL-neutralizing activity was slightly higher for cell line-adapted HIV infection of the H9 T cell line (up to 64% at 50 microg/ml). However, this effect was specific for H9 cells since MBL did not neutralize cell line-adapted virus infection of PBMCs, HIV PI infection of the GHOST cell line, or VSV pseudotyped with HIV gp160 from cell line-derived virus or PI. In contrast to its low activity in neutralization assays, MBL efficiently bound infectious HIV PI and opsonized HIV PI for uptake by monocytic cells. These results show that both PI and cell line-adapted HIV, despite binding of MBL, are relatively resistant to neutralization by levels of MBL normally present in serum. However, binding and opsonization of HIV by MBL may alter virus trafficking and viral-antigen presentation during HIV infection. PMID- 15117455 TI - Fine specificity and cross-clade reactivity of HIV type 1 Gag-specific CD4+ T cells. AB - Despite growing evidence that HIV-1-specific CD4(+) T helper (Th) cells may play a role in the control of viremia, discrete Th cell epitopes remain poorly defined. Furthermore, it is not known whether Th cell responses generated using vaccines based on clade B virus sequences will elicit immune responses that are effective in regions of the world where non-clade B viruses predominate. To address these issues we isolated CD4(+) T cell clones from individuals with vigorous HIV-1-specific Th cell responses and identified the minimum epitopes recognized. The minimum peptide length required for induction of CD4(+) T cell proliferation, IFN-gamma secretion, and cytolytic activity ranged from 9 to 16 amino acids in the five epitopes studied. Cross-clade recognition of the defined epitopes was examined for variant peptides from clades A, B, C, D, and AE. Over half the variant epitopes (17 of 32) exhibited impaired recognition, defined as less than 50% of the IFN-gamma secretion elicited by B clade consensus sequence. There was no evidence for antagonistic activity mediated by the variant peptides, and despite strong responses there was no escape of autologous virus from Th responses in the epitopes we studied. Abrogated recognition of variant CD4(+) T cell epitopes presents a potential obstacle to vaccine development. PMID- 15117457 TI - Determination of HIV type 1 CRF01_AE gag p17 and env-V3 consensus sequences for HIV/AIDS vaccine design. AB - A molecular epidemiological study of the gag p17 and env-V3 regions on HIV infected drug users and blood donors was carried out in northern Thailand from 1998 through 2002 to determine the predominant subtype and consensus sequence (CS) for circulating HIV-1 strains. CRF01_AE was concluded to be a predominant strain and the nucleotide CSs in gag p17 and env-V3 showed only 1.26% and no difference from CS in the Los Alamos database, respectively. Our env-V3 CS was identical to the previously published CSs, suggesting that the CS was very conserved from 1990 through 2002 in Thailand. Gag p17 and env-V3 nucleotide sequences of seroconvertors in our subjects were quite similar to the CS and conserved for at least 9 and 6 years postinfection, respectively. These results suggest that the CS approach to the HIV-1 antigen design could overcome HIV diversity and help us develop an effective HIV/AIDS vaccine. PMID- 15117458 TI - HIV-1 CRF01_AE in intravenous drug users in Hanoi, Vietnam. AB - To investigate the molecular epidemiology of HIV-1 among intravenous drug users (IDUs) in Hanoi we collected 17 samples from individuals living in 12 locations in and around Hanoi. The HIV-1 env V3 and gag p17 regions were directly sequenced from the proviral PBMC population. The majority of the IDUs were infected with HIV-1 CRF01_AE and one individual carried a p17/V3 CRF01/subtype C recombinant. The CRF01 viruses found among these individuals did not seem to be directly epidemiologically linked to each other. The sequences were, however, related to previously reported CRF01 sequences from Vietnam and China. Thus, IDUs in Hanoi seem to have derived their infections in Vietnam, but not from the same source. The discovery of the CRF01/C recombinant shows that new viral forms easily can be generated in IDU transmission chains. PMID- 15117459 TI - Are fusion inhibitors active against all HIV variants? AB - Genetic sequence alignment of the transmembrane region from HIV-1 group O and HIV 2 isolates was performed to examine their potential susceptibility to fusion inhibitors enfuvirtide (T-20) and T-1249. A high genetic diversity within the HRI and HR2 domains was found, which should compromise any antiviral effect of T-20 on HIV-2 and HIV-1 group O viruses. However, conserved sequences in the gp41 regions from HIV-1 group O involved in T-1249 susceptibility might result in a much broader antiviral effect of T-1249 on HIV-1 variants. In contrast, genetic diversity in those regions make unlikely any activity of these compounds on HIV 2. PMID- 15117460 TI - Tularemia in otolaryngology: a forgotten but not gone disease and a possible sign of bio-terrorism. PMID- 15117461 TI - Microbiology of cerumen in patients with recurrent otitis externa and cases with open mastoidectomy cavities. AB - This study investigated the common flora of human cerumen in patients with recurrent otitis externa, and subjects who had been operated on and had an open mastoidectomy cavity from chronic otitis media. Cerumen samples were collected from three groups; group A (n = 20) consisted of patients with recurrent otitis externa, group B (n = 20) consisted of patients with an open cavity and group C (n = 30) consisted of healthy subjects. The mean of the microbial count was 3.4 x 10(4) in group A, 3.08 x 10(4) in group B and 2.48 x 10(4) in group C. The most commonly isolated microorganism from the three groups was Staphylococcus epidermidis. No growth was observed in five cases (25 per cent) in group A and in three cases (10 per cent) in group C. In group B antimicrobial growth was observed in all samples. In 46 (65 per cent) of the cerumen samples, the isolates were monomicrobial and 24 (35 per cent) of the cerumen samples were polymicrobial. The isolates were polymicrobial in 65 per cent of group A, 20 per cent in group B and 23.3 per cent in group C. In the process of investigating the microbial flora of cerumen in all the three groups, microbial growth was observed from all the samples from patients with an open cavity, unlike the other groups, and it was determined that the group with recurrent external otitis had the most abundant microbial flora. PMID- 15117462 TI - Early experience with the Mills sleeve prosthesis for reconstruction of the incus long process. AB - This paper describes the Mills sleeve technique of ossicular reconstruction for defects of the incus long process and reviews the results of 27 procedures. A retrospective review of operations was performed by eight otologists. The results are compared with those from three other reconstruction techniques (cortical bone graft, cortical bone sleeve and incus autograft). Prostheses were supplied to surgeons who expressed an interest. Pre- and post-operative audiological data forms were analysed for each case along with a questionnaire about use of the prosthesis. The mean post-operative air bone gap (ABG) was compared with results from cortical bone sleeve, incus autograft and simple cortical bone graft reconstruction cases previously performed by the senior author. Twenty-seven procedures were performed. Closure of the ABG to within 10 dB was achieved for 44.4 per cent of Mills sleeve cases compared with 44.7 per cent for the cortical bone sleeve, 52.9 per cent for ossicular and 26.9 per cent for cortical bone grafts. The responses to a questionnaire sent to participating surgeons are discussed. For the current follow-up period (three months to three years) the Mills sleeve prosthesis appears to be safe and easy to use with audiological results at least as good as other reconstructive techniques. PMID- 15117463 TI - Concomitant nasal procedures in endoscopic dacryocystorhinostomy. AB - Dacryocystorhinostomy (DCR) is the preferred treatment for lacrimal duct blockage. Endoscopic DCR has been practised increasingly in recent years as it avoids a facial scar and can be performed as a day procedure. Recent improvements in endonasal surgical technique led to success rates of up to 90 per cent. However, the endonasal approach often requires septal or turbinate surgery to optimze access to the lacrimal area. The incidence of concomitant procedures was investigated in 256 patients undergoing endonasal nonlaser DCR. In this study 55 out of 256 patients (21.5 per cent) required additional endonasal procedures to improve access to the lacrimal area. It is therefore advisable that otolaryngologists are involved in this procedure. PMID- 15117464 TI - Upper airway surgery benefits patients with obstructive sleep apnoea who cannot tolerate nasal continuous positive airway pressure. AB - Nasal continuous positive airway pressure (CPAP) is the mainstay of treatment for patients with moderate to severe obstructive sleep apnoea (OSA). However, tolerance and compliance are poor. An audit using the Christchurch Hospital ORL surgery database identified patients who underwent upper airway surgery for OSA. Tracheostomy and bimaxillary advancement patients were excluded. Adults with moderate to severe OSA (Desaturation Index (DI) >10 n.h(-1)), who had failed a trial of nasal CPAP, and had pre-operative and post-operative sleep study data were identified. Objective (DI) and Subjective (Epworth Sleepiness Score (ESS)) outcome measures were recorded. The database identified 69 patients who underwent surgery for snoring or OSA; of these, 25 patients formed the study group. Sixteen out of 25 improved (64 per cent) after surgery, seven out of 25 showed no change (28 per cent), two patients (eight per cent) showed deterioration in their DI. Forty-eight per cent of patients had >50 per cent post-operative improvement in DI. Fourteen out of 25 (56 per cent) had a post-operative DI <20 n.h(-1). Seven out of 25 (28 per cent) had a post-operative DI <10 n.h(-1). Upper airway surgery has a role in the management of selected patients with OSA who cannot tolerate nasal CPAP. PMID- 15117465 TI - Paediatric obstructive sleep apnoea: is a polysomnogram always necessary? AB - Obstructive sleep apnoea (OSA) is a common entity in children, most present with sleep disturbances such as snoring, choking during sleep, enuresis, restless sleep, or apnoeic spells. Other symptoms include poor school performance, hyperactivity, failure to thrive, heart failure and cor pulmonale. Most authors would concur that the polysomnogram (PSG) is the gold standard for the diagnosis of OSA, and that adenotonsillectomy is the surgical procedure of choice, with high curative rates and relatively low morbidity. Close post-operative monitoring of all children with OSA cannot be over-emphasized. The focus has been, traditionally, to anticipate post-operative airway and respiratory complications in this group of children. We present 73 children with clinical OSA and 36 children with proven OSA on PSG, with only one child having respiratory complications (mixed apnoea), and all with uneventful recovery. In view of our low complication rates, low post-operative morbidity, cost and facility factor, the need for a mandatory overnight PSG pre-operatively is questioned, and clinical criteria for performing a PSG preoperatively are suggested. PMID- 15117466 TI - Management of laryngeal amyloidosis. AB - Amyloidosis of the upper aerodigestive tract is rare. However, the larynx is the commonest site within the upper airways to be affected. Three cases of uncomplicated primary amyloidosis of the larynx and one case of laryngeal amyloidosis with more generalized dissemination throughout the head and neck are presented. In this article the different presentations of laryngeal amyloidosis and principles of management of this condition at that site are highlighted. PMID- 15117467 TI - Quality of life and functional evaluation after supracricoid partial laryngectomy with cricohyoidoepiglottopexy in Mexican patients. AB - This study assessed the functional results in patients treated primarily through supracricoid partial laryngectomy (SCPL) with cricohyoidoepiglottopexy (CHEP). Fifteen patients with a diagnosis of epidermoid carcinoma of the glottis region admitted to the Instituto Nacional de Cancerologia (Mexico) between June 2001 and September 2002 were studied. Three patients were at stage I, five at stage II, six at stage III, and one at stage IV. Both cricoarytenoid units were preserved in 12 patients, and only one in three. Each case was assessed through the clinical grading postoperative aspiration (CGPA) scale, the performance status scale for head and neck cancer (PSS-HNC), and the Karnofsky Performance Scale (KPS). Likewise, voice quality of the patients was assessed regarding tone and intensity using the SpeechViewer version 1 (IBM) and data were obtained with the Cool Edit 2000 software. Twelve patients received phoniatric rehabilitation and three were left without rehabilitation. The average time for decannulation was 12 days and 23 days for removal of the nasogastric catheter. The degree of aspiration was 0 in four patients and one in 11. According to PSS-HNC, the mean for normalcy in the diet was 95 and the mean for those eating in public was 91. Intelligibility reached an average of 90. Karnofsky's assessment was related to the disease and not to the treatment, as it remained at 100 per cent in most patients and was never below 80 per cent. The mean intensity of quality of voice was -18 dB below normal; however, the mean frequency was 243.7 Hz. SCPL and CHEP allows the preservation of the basic function of the larynx; however, a clear alteration in voice occurs after the procedure, although normal frequency is kept when both arytenoids are preserved. Likewise, preservation of both arytenoids shortens the time needed for cannula and feeding catheter removal. PSS-HNC, Karnofsky, and CGPA assessments demonstrated that patients can reach an almost normal bio-psycho-social integration. It is recommended that all patients be subjected to phoniatric rehabilitation. PMID- 15117468 TI - Determination of the changes in the hypoglossal nerve function after suspension laryngoscopy with needle electromyography of the tongue. AB - The purpose of this study was to determine changes in the hypoglossal nerve function after suspension laryngoscopy with needle electromyography of the tongue. This study also attempted to determine the possible relationship between the predictive factors of intubation difficulty by using the intubation difficulty scale, which was introduced by Adnet et al., duration of suspension laryngoscopy and changes in hypoglossal nerve function after suspension laryngoscopy. The study was performed on 39 patients who underwent suspension laryngoscopy for benign glottic pathology. Pre-operative airway assessment was evaluated by the intubation difficulty scale and the duration of suspension laryngoscopy was recorded. Needle electromyography of the tongue was performed three or four weeks after the suspension laryngoscopy. After needle electromyography of the tongue, increased polyphasia was found in 13 patients (33 per cent), bilaterally in three of them. The interference pattern was reduced in two of these 13 patients. There was no statistically significant difference in predictive factors of intubation difficulty and the duration of the operation between these 13 patients with increased polyphasia and the remaining 26 patients with completely normal electromyography findings. These findings show that, in spite of normal clinical tongue function, subclinical changes can be detected by needle electromyography of the tongue after suspension laryngoscopy. PMID- 15117469 TI - Role of repeat fine needle aspiration cytology in head and neck lesions: preliminary study. AB - In this preliminary prospective study the value of repeating fine needle aspiration cytology (FNAC) in patients with head and neck lesions was investigated. Few reports exist on the significance of repeating the procedure in head and neck patients. Fifty-seven patients have been sampled twice for the first and second (repeat) FNAC. The second aspirate was performed in the operating theatre under general anaesthesia prior to a surgical procedure. The cytological results were compared with the histology of the 57 resected lesions. It was found that the overall diagnostic results improved after repeating the FNAC. It can be concluded that repeating FNAC is useful and should be considered under some circumstances, especially in the case of non-diagnostic cervical lymph node aspirates. PMID- 15117470 TI - Use of a ureteric pigtail stent as a self-retaining frontal sinus stent. AB - There have been many frontal sinus stents described in the medical literature. Here we describe the first application of a ureteric pigtail stent to an extensively operated frontal sinus and lateral mucocele cavity. There are two properties of the design of this stent that make it ideal for this case. Firstly, it is self-retaining, eliminating the need for fixation sutures. Secondly, the stent has a considerable available length (220-300 mm between pigtails), allowing ventilation of a distant mucocele cavity. PMID- 15117471 TI - Far advanced otosclerosis and intractable benign paroxysmal positional vertigo treated with combined cochlear implantation and posterior semicircular canal occlusion. AB - This paper presents a combined procedure for the management of intractable benign paroxysmal positional vertigo (BPPV) and profound hearing loss in a patient with far advanced otosclerosis. The procedure comprised of a posterior semicircular canal occlusion and cochlear implantation as one combined procedure in the same ear. The combined approach added little to the operative morbidity and proved effective in this patient's management. A search of the literature reveals this to be a unique case. PMID- 15117472 TI - Nasal polyposis: unusual indication for tracheostomy. AB - A rare case of nasal polyposis requiring urgent tracheostomy is reported. A 70 year-old male presented to the hospital with stridor. He had a 10-year history of nasal obstruction. A large proliferative mass was found to be occupying both the nasal cavities. An emergency tracheostomy and biopsy of the lesion was performed. A computerized axial scan (CT) demonstrated that the lesion occupied the pharynx reaching up to the laryngeal inlet. To our knowledge this is the first case reported in the literature. Upper airway obstruction from nasal polyps is uncommon but can cause significant morbidity if not appropriately managed. PMID- 15117473 TI - Masquerade syndrome: sebaceous carcinoma presenting as an unknown primary with pagetoid spread to the nasal cavity. AB - Sebaceous carcinoma of the eyelid is an uncommon tumour with unusual modes of presentation. It can remain occult at the primary site, without producing any mass, masquerading as chronic blepharoconjunctivitis, while setting up metastases in the regional lymph nodes especially in the pre-auricular group. We report here a case that not only masqueraded as chronic blepharoconjunctivitis with nodal metastases from an 'unknown primary' in the neck, but whose tumour spread in a pagetoid manner along the nasolacrimal duct producing a nasal tumour that was believed to be the 'unknown primary'. This case emphasizes the need for ophthalmologists, ENT surgeons and pathologists to keep sebaceous carcinoma in mind while evaluating patients with chronic blepharoconjunctivitis and cervical node metastases from 'unknown primary'. Histological clues for picking up a sebaceous carcinoma at a metastatic site include a tumour with comedo or ductal growth pattern and intracytoplasmic lipid. PMID- 15117474 TI - Spontaneous enophthalmos: silent sinus syndrome. AB - Spontaneous enophthalmos unrelated to trauma or surgery is rare. The term 'silent sinus syndrome' has been used to describe this process where, in particular, there is an absence of any sino-nasal symptoms. The enophthalmos and hypoglobus that occurs in these subjects is caused by atelectasis of the maxillary antrum, which itself appears to be due to chronic maxillary hypoventilation. We report a case of silent sinus syndrome that arose following insertion of a nasogastric tube. Whilst acute paranasal sinusitis is a well-described sequela of nasal intubation, this association with a rare, and as yet unexplained, phenomenon may go some way to explain its aetiology. PMID- 15117475 TI - Surgery-induced thyroiditis following laryngectomy. AB - A 77-year-old euthyroid man developed atrial fibrillation on the fourth post operative day following a total laryngectomy and right radical neck dissection including a hemi-thyroidectomy for a squamous cell carcinoma of the larynx with nodal metastases. The episode of atrial fibrillation coincided with an elevated serum free thyroxine of 3.36 ng/dl (43.3 pmol/L) (reference values: 0.71-1.85 ng/dl or 9.1-23.8 pmol/L), normal total triiodothyronine of 104 ng/dl (1.6 nmol/L) (reference values: 46-137 ng/dl or 0.7-2.1 nmol/L) and a suppressed thyroid-stimulating hormone (TSH) of 0.05 mIU/L (reference values: 0.35-5.0 mIU/L). These values, in relation to those prior and subsequent, suggested a surgery-induced thyroiditis. The limited literature about this controversial entity is reviewed. PMID- 15117476 TI - Gastrointestinal stromal tumour of the pharynx. AB - This documents the case of a 55-year-old female presenting with a solitary polypoidal tumour of the pharynx. Histological examination revealed features consistent with a gastrointestinal stromal tumour. Although well described elsewhere in the gastrointestinal tract, from our literature search, this is the first reported case of such a tumour occurring in the pharynx PMID- 15117477 TI - Follicular dendritic cell sarcoma arising from the hypopharynx. AB - Follicular dendritic cell (FDC) tumours have been described recently as malignant tumours arising from accessory cells of the lymph nodes. They are rare tumours with fewer than 70 cases occurring worldwide. They usually present in cervical or abdominal lymph nodes, with very few occurring extranodally. We present the first case of an FDC tumour to occur in the hypopharynx with simultaneous cervical node metastases. The pathology is discussed and the literature reviewed. PMID- 15117478 TI - Left atrial myxoma manifesting as acute isolated vertigo. PMID- 15117479 TI - Revolutions and revelations. PMID- 15117480 TI - Efficacy of combined photodynamic and hyperthermic therapy with a new light source in an in vivo osteosarcoma tumor model. AB - OBJECTIVE: In this study, we investigated the efficacy of Super Lizer (SL) as a new light source in photodynamic therapy (PDT) with hyperthermia in an in vivo osteosarcoma tumor model. MATERIALS AND METHODS: Nude mice in three study groups (PDT only, PDT with hyperthermia in low energy, and PDT with hyperthermia in high energy) and three control groups (no treatment, photosensitizer only, and hyperthermia only) were implanted subcutaneously with human osteosarcoma cells and injected with a photosensitizing hematoporphyrin derivative (HPD) at a total dose of 10 mg/kg, in all study groups and in control group 2. At 72 h after light treatment, mice were sacrificed. RESULTS: The tumor volume growth rates in the heat-only (1.50) and PDT-only (1.40) groups were significantly lower than the growth rate in the no-treatment group (1.82). Further, the tumor volume growth rate in the PDT with hyperthermia in high-energy group (1.19) was significantly lower than in the heat- or PDT-only groups. CONCLUSION: Although non-laser PDT, including SL-PDT, may be beneficial only in the treatment of superficial tumors because of limited light penetration, PDT combined with hyperthermia may extend the utility of PDT in antitumor treatment. The use of SL as a new light source in PDT may significantly advance antitumor therapy due to its simplicity, ease, and cost benefit. PMID- 15117481 TI - Bactericidal effect of a 980-nm diode laser in the root canal wall dentin of bovine teeth. AB - OBJECTIVE: The aim of this in vitro study was to investigate the antibacterial depth effect of continuous wave laser irradiation with a wavelength of 980 nm in the root canal wall dentin of bovine teeth. BACKGROUND DATA: The long-term success of an endodontic therapy often fails due to remaining bacteria in the root canal or dentin tubules, which cannot be sufficiently eliminated through the classical root canal preparation technique nor through rinsing solutions. MATERIALS AND METHODS: A total of 102 slices of bovine root dentin of different thicknesses (100, 300 and 500 micro m) were prepared. The samples were inoculated from one side with 5 micro L of an enterococcus faecalis suspension of defined concentration. Four slices per slice thickness served as a control group; the rest of the 30 slices per thickness were subjected to laser irradiation - 10 each of these slices were irradiated with distal outputs of 1.75, 2.3, and 2.8 Watts (W). After drying them for 30 sec, the back of the inoculated dentin slice was irradiated for 32 seconds with a 200- micro m fiber optical waveguide under constant movement of the fibers. The remaining bacteria were then detached in NaCl under vibration. The eluate produced by this was - taking account of the degree of dilution - plated out on sheep blood agar plates. After 24 h of incubation, the grown bacterial colonies were able to be counted out and evaluated. By doing so, they were compared with the non-irradiated, but otherwise identically treated control group. RESULTS: With a slice thickness of 100 micro m, the 980-nm diode laser achieved a maximum bacterial reduction of 95% at 1.75 W, 96% at 2.3 W, and 97% at 2.8 W. With a slice thickness of 300 micro m, a maximum of 77% of the bacteria was destroyed at 1.75 W, 87% at 2.3 W, and 89% at 2.8 W. The maximum bacterial reduction with a slice thickness of 500 micro m was 57% at 1.75 W, 66% at 2.3 W, and 86% at 2.8 W. CONCLUSION: The results of this research show that the 980-nm diode laser can eliminate bacteria that have immigrated deep into the dentin, thus being able to increase the success rate in endodontic therapy. PMID- 15117482 TI - The comparison of effects between pulsed and CW lasers on wound healing. AB - OBJECTIVE: In order to evaluate the effects of pulsed continuous wave (CW) laser and detect the role of wound healing in rats using both pulsed and CW 635-nm low level laser therapy (LLLT), a pilot study was undertaken. BACKGROUND DATA: Some acceleration effects of wound healing on animals were found after treatment using various lasers with CW. There are other reports, however, using pulsed CW laser to evaluate the effects of wound healing in rats. MATERIALS AND METHODS: An elliptic wound was created aseptically with a scalpel on the shaved back of the rats after anesthesia. The rats treated were restrained in a Plexiglas cage without anesthesia during the laser irradiation period. An Erchonia pulse laser (635 nm) was used in the experiment. The laser beam was delivered through an expander. The percentage of relative wound healing was calculated. RESULTS: The percentage of relative wound healing was 4.32 in 100 Hz, 3.21 in 200 Hz, 3.83 in 300 Hz, 2.22 in 400 Hz, 1.73 in 500 Hz and 4.81 in CW. CONCLUSION: LLLT using pulsed, CW laser at the appropriate dosimetry and frequency can provide acceleration in wound healing in rats. The 100-Hz frequency had a better effect than other pulse frequencies used in the study. The effects of treatment using CW laser was higher than pulse frequency. The frequency of pulsed CW laser was not found to increase wound healing in rats compared with normal CW laser, as reported in our previous studies. PMID- 15117483 TI - Dose and wavelength of laser light have influence on the repair of cutaneous wounds. AB - OBJECTIVE: The objective of the present study was to compare histologically the effect of GaAlAs (lambda 830 nm, phi approximately 2 mm(2), 35 mW) and InGaAlP (lambda 685 nm, phi approximately 2 mm(2), 35 mW) lasers, alone or in association with doses of 20 or 50 J/cm(2) on cutaneous wounds in the dorsum of the Wistar rat. BACKGROUND DATA: The healing time of surgical wounds is of extreme importance and it is usually associated with a post-operative period free of infection and with less pain and inflammation. MATERIALS AND METHODS: Sixty Wistar rats were divided into seven groups: Group I - control (non-irradiated); Group II - lambda 685 nm, 20 J/cm(2); Group III - lambda 830 nm, 20 J/cm(2); Group IV - lambda 685 nm and lambda 830 nm, 20 J/cm(2); Group V - lambda 685 nm, 50 J/cm(2)); Group VI - lambda 830 nm, 50 J/cm(2); and Group VII - lambda 685 nm and 830 nm, 50 J/cm(2). The animals were sacrificed 3, 5, and 7 days after surgery. RESULTS: Light microscopic analysis using H&E and Picrosirius stains showed that, at the end of the experimental period, irradiated subjects showed increased collagen production and organization when compared to non-irradiated controls. Inflammation was still present in all groups at this time. CONCLUSION: Group IV (lambda 830 nm and lambda 685 nm, 20 J/cm(2)) presented better results at the end of the experimental period. It is concluded that low-level light therapy (LLLT) can have a positive biomodulatory effect on the repair of cutaneous wounds. PMID- 15117484 TI - Management of oral leukoplakia by laser surgery: relation between recurrence and malignant transformation and clinicopathological features. AB - OBJECTIVE: The aim of the present study was to assess the clinical usefulness of laser surgery for oral leukoplakia. BACKGROUND: Recurrence and/or malignanT transformation of oral leukoplakia have occasionally been observed following laser surgery. It is reported that the rate of recurrence was 7.7-38.1%, while malignant transformation was 2.6-9%. MATERIALS AND METHODS: A total of 154 oral leukoplakias from 116 patients were treated in our department. Comparisons with the rate of recurrence, malignant transformation and clinical futures, epithelial dysplasia, location, and treatment procedure were examined. To evaluate recurrence and malignant transformation, the cases were restricted to those with a minimum follow-up of 6 months. RESULTS: A total of 97 lesions fulfilled this criterion. The rate of recurrence had no association with the location or the epithelial dysplasia, although it differed with the treatment procedure. The rate of recurrence in laser surgery was approximately 29%. Malignant transformation was observed in four of 97 lesions. They were observed in 13.6% of tongue cases and 1.8% of gingival cases. Three of four lesions were treated with excision surgery, and the remaining one treated with laser vaporization. Only 1.2% malignant transformation in laser surgery was revealed. CONCLUSION: Laser excision is suitable for leukoplakia cases on non-keratinized epithelia (i.e., the tongue and buccal mucosa), while laser vaporization is suitable for the gingival cases. Management of oral leukoplakia prevents not only recurrence and malignant transformation, but also postoperative dysfunction. We believe that laser surgery is an excellent procedure. PMID- 15117485 TI - SEM analysis of dentin treated with the Er:YAG laser: a pilot study of the consequences resulting from laser use on adhesion mechanisms. AB - OBJECTIVES: The purpose of this study was to compare the morphological characteristics of lased dentinal surfaces obtained by chemical etching, which was applied either before or after laser treatment. The adhesive interface after the application of an adhesive-composite system on the above mentioned dentinal surfaces was also evaluated. MATERIALS AND METHODS: Eighteen caries-free permanent molars were transversally cut to obtain the dentinal surfaces. Ten of these surfaces were divided into three main groups according to their preparation modality: laser only, laser followed by chemical etching, and chemical etching before laser. The Er:YAG laser was used with two different energy outputs: 200 and 400 mJ at 1 Hz. The eight remaining surfaces were divided into two groups and followed the same preparation as the first three groups. The only difference was a change in frequency: 4 Hz was used for treating these surfaces. An adhesive system (Scotchbond 1, 3M), a thin layer of flowable composite (Tetric Flow, Vivadent), and a regular composite (Z250, 3M) were applied to each dentinal surface and then separately light cured. In order to evaluate the dentine restoration interface, the samples were cut longitudinally and prepared for SEM observation. RESULTS: The dentinal surfaces that were etched by the laser treatment did not demonstrate the characteristics necessary for guaranteeing a good bond with the adhesive materials. A closer observation of the dentine restoration interface revealed some micro-fractures located just below the lased dentinal surface. CONCLUSION: The decision to use the Er:YAG laser as an alternative to conventional techniques of acid etching is currently arguable. Given that the available adhesive systems require acid conditioning, it would not be advisable to skip this operative step. PMID- 15117486 TI - Dentin evaluation after Nd:YAG laser irradiation using short and long pulses. AB - OBJECTIVE: Several reports have demonstrated the advantages of using the Nd:YAG laser to reduce dentin permeability by melting the dentin surface. A comparative study using different pulse durations can be useful to obtain further information about the laser-hard tissue interaction. MATERIALS AND METHODS: The present study pursues the evaluation of the morphological and chemical changes in human dentin surface resulting from Nd:YAG laser (lambda = 1064 nm) irradiation, with a total energy of 0.9 J distributed in 1, 2, 3, and 6 pulses with different pulse durations to promote surface melting and dentinal tubule occlusion. After irradiation, the samples were submitted to scanning electron microscopy (SEM) analysis for morphological study and energy-dispersive spectrometry (EDS) analysis for evaluation of the concentration of calcium and phosphorous in the melted layer. RESULTS: SEM analysis of the irradiated dentin surface showed surface structural changes due to laser irradiation, where the morphological changes are dependent on the laser pulse duration. EDS analysis showed an increase of calcium and phosphorous concentrations after Nd:YAG laser exposure, but no correlation with the number of pulses or pulse duration was found. CONCLUSION: Our results suggest that longer interaction times resulted in more evident effects with more melted substrate than shorter pulses, and in both cases the resultant melted layer contains a greater concentration of inorganic substances than non-irradiated dentin. PMID- 15117487 TI - Study of microleakage at class V cavities prepared by Er:YAG laser using rewetting surface treatment. AB - OBJECTIVE: This study was conducted to analyze microleakage in Class V cavity preparation, using rewetting (or not) just after burr or Er:YAG laser preparation of enamel and dentin walls in permanent teeth. BACKGROUND DATA: Several studies reported microleakage around composite restorations when cavity preparation was done or treated by Er:YAG laser. As the hybridized laser is removed when this laser is used to cut dental hard tissue, there is a need for new materials or techniques to minimize gaps and microleakage. RESULTS: Primer solution showed significant effect in enamel and dentin, at the level of 5%, when Er:YAG laser was used as a cutting tool. Using primer solution after phosphoric acid in preparations with the laser, microleakage was similar in degree to when cavities were prepared with the burr. CONCLUSION: Re-wetting surface just after Er:YAG irradiation and chemical treatment with phosphoric acid using HEMA aqueous solution seems to improve the quality of bioattachment between the adhesive system and enamel/dentin, showing similarities between restoration behaviors independently of the cutting tool, whether burr or laser. PMID- 15117488 TI - Complications associated with cervical endoscopic discectomy with the holmium laser. AB - OBJECTIVES: Our aim was to determine the rate of surgical complications associated with cervical endoscopic discectomy (CED). BACKGROUND DATA: There are no studies that state the degree of complications after CED. MATERIALS AND METHODS: Forty-one patients underwent CED with holmium laser. RESULTS: Two out of 41 patients incurred vascular compromise during the procedure. One patient developed recurrent laryngeal nerve damage. One patient developed discitis, and two patients complained of a "clicking" sensation postoperatively. CONCLUSIONS: Although CED has a relatively high success rate, there is a 15% rate of complications associated with the procedure. Most of the complications were minor (such as vascular compromise, recurrent laryngeal nerve injury, and postoperative "clicking" sensations), but there was one case of severe discitis, and there is the potential of serious complication from both vascular compromise and neural injury. PMID- 15117489 TI - Effects of low-intensity polarized visible laser radiation on skin burns: a light microscopy study. AB - OBJECTIVE: This study was carried out to investigate the influence of low intensity polarized visible laser radiation on the acceleration of skin wound healing. BACKGROUND DATA: Low-level laser therapy (LLLT) at adequate wavelength, intensity, and dose can accelerate tissue repair. However, there is still unclear information about light characteristics, such as coherence and polarization. Some studies indicate that linearly polarized light can survive through long propagation distance in biological tissue. MATERIALS AND METHODS: Three burns about 6 mm in diameter were created on the back of rats with liquid N(2). Lesion "L(//)" was irradiated by He-Ne laser (lambda = 632.8 nm), D= 1.0 J/cm(2), with linear polarization parallel to the spinal column of the rat. Lesion "L(inverted v)" was irradiated using the same laser and dose, but the light polarization was aligned perpendicularly to the relative orientation. Lesion "C" was not irradiated in order to be considered as control. The animals were sacrificed at day 3-17 after lesion creation. Samples were collected and prepared for histological analysis. RESULTS: Histological analysis showed that the healing of irradiated wounds was faster than that of non-irradiated wounds. Moreover, it was observed that skin wound repair is dependent on polarization orientation with respect to a referential axis as the animal's spinal column. Consequently, "L(//)" was completely healed after 17 days, whereas "L (perpendicular) " showed a moderate degree of healing after the same period. CONCLUSIONS: These results indicate that the relative direction of the laser polarization plays an important role in the wound healing process when highly coherent He-Ne laser is used. PMID- 15117491 TI - Laser literature watch. PMID- 15117492 TI - Cover up or cool it? Sexual intercourse during therapy for bacterial sexually transmitted infections--a discussion of evidence for efficacy of condom use preventing transmission during an acute bacterial STI. AB - Effective treatment of sexually transmitted infections (STIs) such as gonorrhoea and chlamydia needs sexual behaviour modification, in addition to antibiotics, to protect the index patient and their sexual partner from re-infection during treatment. This may mean advice to avoid sex, or to use condoms for intercourse, until all current sexual partners have completed treatment. This article discusses the effectiveness of condoms as re-infection protection during therapy of gonorrhoea and chlamydial infection. Indirect evidence from studies on primary prevention indicates that, although physically, condoms are impermeable to STI pathogens, the risk of failure depends on the experience of the user and the frequency of use. Health care workers may over-estimate the protective value of condoms in this situation. Patients should be made aware of the risk of infection present, even in experienced condom users, if they choose to continue protected sexual intercourse during therapy for acute bacterial STIs. PMID- 15117493 TI - Hepatitis C and HIV co-infection. AB - HIV accelerates progression of hepatitis C virus (HCV)-related liver disease. There are conflicting data on the effect of HCV on the risk of HIV progression and CD4 response to highly active antiretroviral therapy (HAART). Long-term prospective cohort studies are clearly required to resolve these issues. The optimal management of the co-infected patient is also unclear. For the co infected patient, the optimal HAART regimen for best immune CD4 recovery and least adverse reactions remains unclear. Unfortunately, current HCV treatment is associated with significant side effects and a considerable proportion of HIV co infected patients are poor candidates for HCV treatment. Better and more effective treatment for HCV (preferably not based on interferon) is urgently required for this group of patients. Patients with good CD4 cell count and with HCV genotypes 2 and 3 are likely to have a reasonable response to treatment. PMID- 15117495 TI - Getting the guidance right: optimizing the quality of the UK national guidelines on sexually transmitted infections and closely related conditions. PMID- 15117496 TI - Specifications for the development of guidelines on the management of sexually transmitted infections and closely related conditions. PMID- 15117497 TI - What is the role of scrotal ultrasound scans in genitourinary medicine? AB - The objective of the study was to review the indications for scrotal ultrasound scans and to assess the impact on patient management. We therefore performed a retrospective analysis. Case notes of all males referred for a scrotal ultrasound between April 1998 and March 2001 were studied. Data were extracted for the following: age, presenting complaints, physical findings on examination, results of a full sexual screen, treatment, ultrasound result and the designation of the person requesting the scan. All data were tabulated and summated using 'Windows Excel' software. One hundred and fifteen men were referred for an ultrasound in this period of time. None had an ultrasound more than once. Of these, 25 subjects could not be included as they either failed to attend for their scan (n=8) or their notes could not be traced (n=17). Twelve subjects were excluded as they did not fit the selection criteria (n=12). Median age was 30 years (Range 19-61 years). The commonest reason for referral was testicular pain (n=43) followed by testicular lump (n=19). The commonest abnormality on examination was an inflamed epididymis (n=18). Forty-two of the ultrasound scans were normal. A testicular mass was detected in only five of the 78 patients (6.4%), of which one was a malignancy (1.2%). A direct referral to a urologist for further management was made in only nine patients. Scrotal ultrasound for pain has limited impact on patient management apart from reassuring a worried patient. However, it remains an important investigation in the management of a suspected testicular lump. PMID- 15117498 TI - Emergency contraception: who are the users? AB - The objective was to describe the demographic and sexual characteristics of clients attending a Sexual Health Clinic for emergency contraception (EC). Information about women attending the Parramatta Sexual Health Clinic (PSHC) who received EC between January 1999 and July 2002 was derived from the clinic database. Age-matched controls were randomly selected. Univariate and logistic regression analysis was performed to establish which factors were associated with use of EC. Two hundred and sixty-seven women requesting EC, and an equal number of controls, were studied. Factors that were independently associated with EC use were being a student, (OR=1.7 [95% CI 1.02-2.69]) and having a regular sexual partner (OR=2.3 [95% CI 1.14-4.73]). Women requiring EC were significantly less likely to have had a sexually transmitted infection (STI) (OR=0.3 [95% CI 0.16 0.60]) or a previous pregnancy (OR=0.2 [95% CI 0.09-0.67]) than controls. We concluded that users of EC are at low-risk for STIs, but need counselling about safer sex. PMID- 15117499 TI - Private prescription costs for sildenafil within the NHS: a telephone survey. AB - Because of the restrictions on prescribing for impotence within the NHS, doctors routinely write private prescriptions for sildenafil. The aim of this study was to determine the variation in cost of a private prescription of four 100 mg tablets of sildenafil. A selection of different pharmacy types within five areas in England was surveyed. We telephoned a total of 86 pharmacies and we were quoted prices ranging from pounds 28.20 to pounds 42.33. There was a significant difference in price between area and between pharmacy type. Best prices are not necessarily found at the major pharmacy chains or hospital pharmacies, as might be expected. NHS doctors and patients need to be aware of this significant difference in cost. PMID- 15117500 TI - Patient assessment of anogenital warts and the success of treatment with home applied therapy. AB - Home treatment with podophyllotoxin or imiquimod are commonly prescribed therapies for anogenital warts. It is important to ascertain if patients are locating all lesions for treatment and if they know when they are clear of them. We set out to assess patients' ability to determine the number and location of their genital warts and compare their observation with that of their examining doctor or nurse. Following instruction on the use of home treatment and being given an instruction leaflet patients were reviewed in four weeks' time. One hundred and fifty-five patients enrolled in the study--31% (48) male, 69% (107) female. At initial assessment 62.5% (30) of male patients and 59.8% (64) of female patients underestimated the extent of their disease: 10.5% (5) of male patients and 10.3% (11) of female patients overestimated their disease burden with some mistaking skin tags for genital warts. At review 29.4% (5) of male patients and 44.4% (20) of female patients still underestimated the extent of their infection. Patients undertaking home treatment for warts not only need detailed instruction on its use but should be reviewed to assess the success of treatment. PMID- 15117501 TI - Retrospective review of the correlation of symptoms, signs and microscopy with the diagnosis of Chlamydia trachomatis in men. AB - We have investigated the correlation of symptoms, signs and microscopy on the detection of Chlamydia trachomatis infection in men attending our clinic. One hundred and eleven men diagnosed with chlamydia over an 11-month period in our clinic were reviewed. Of these, 70 (63%) were symptomatic and 41 (37%) were asymptomatic. 84% (59/70) of symptomatic and 51% (21/41) of asymptomatic patients met the UK clinical criteria for diagnosis of non-gonococcal urethritis (NGU) and received treatment appropriate for chlamydia and NGU at their initial clinic visit. Our findings show that 19% (21/111) of men with Chlamydia trachomatis infection would leave the clinic untreated if a urethral smear preparation was not performed on those who were clinically asymptomatic at presentation. We feel that this is a sufficient argument to warrant continuing the current practice of testing asymptomatic men attending GUM clinics for NGU. PMID- 15117502 TI - Evaluation of possible effects of continued drug use on HIV progression among women. AB - Data from a prospective, multi-centred study of HIV infection in women (HIV Epidemiology Research Study [HERS]) was analysed to investigate the effect of continued injection drug use behaviours on progression to AIDS. All women enrolled in the HERS had at enrollment and at six-month intervals, a face-to-face interview which included specific injection drug use, a physical exam, and specimen collection that included T-cell subset analysis and HIV plasma RNA detection. Six hundred and thirty-nine HIV-infected women contributed 3021 person years of observation during 7.25 years of follow-up, and 299 of these women progressed to AIDS (46.8%). In multivariable analysis, there was no significantly increased risk of progression to AIDS for women reporting pre-baseline injection drug use [hazard ratio (HR)=1.07 (0.78, 1.47)] or reported injection drug use during follow-up [HR=0.89 (0.66, 1.21)] compared with never injecting. In a separate multivariable-model, comparing women who reported no injection in past six months to active injection drug users, the frequency of injection during the previous six months measured by daily injection [HR=0.97 (0.61, 1.55)] or less than daily injection [HR=0.84 (0.54, 1.33)] was not associated with progression to AIDS. Being in drug treatment was independently associated with a slower progression to AIDS [HR=0.41 (0.28, 0.59)]. Neither injection drug use, nor frequency of injection drug use was associated with progression to AIDS among HIV infected women. Initiation of antiretroviral therapy among drug users should be based on readiness for treatment rather than concern about faster progression. PMID- 15117503 TI - Response of HIV-infected patients with asymptomatic syphilis to intensive intramuscular therapy with ceftriaxone or procaine penicillin. AB - The objective of this prospective pilot study was to evaluate the response of HIV infected patients with asymptomatic syphilis to one of two intensive antibiotic treatment regimens. Thirty-one HIV-infected patients with serum rapid plasma reagin titre > or =1:4 and no clinical findings of syphilis were randomized to receive daily intramuscular injections of ceftriaxone or procaine penicillin (plus oral probenecid) for 15 days; 24 returned for follow-up study. Seven of 10 (70%) procaine penicillin-treated patients and 10 of 14 (71%) ceftriaxone-treated patients had a > or =4-fold decline in RPR (P=0.94); two penicillin-treated and one ceftriaxone-treated patient relapsed. Two patients failed ceftriaxone therapy. Three penicillin-treated, and two ceftriaxone-treated patients were serofast. Serological responses were similar in those patients with and without asymptomatic neurosyphilis. There was no difference in the serologic response to daily treatment with ceftriaxone vs that with procaine penicillin plus probenecid; both treatments were associated with comparatively high rates of serological non-response and relapse. PMID- 15117504 TI - Pharmacy clerks' prescribing practices for STD patients in Porto Alegre, Brazil: missed opportunities for improving STD control. AB - STDs are a significant public health problem in Brazil. A primary control strategy is the immediate treatment of symptomatic individuals. When services are unavailable, STD patients seek care in alternative settings. Probably the most frequently used settings are commercial pharmacies, where pharmacy clerks provide treatment, although Brazilian law prohibits selling antibiotics without prescription. Our objective was to evaluate prescribing practices by pharmacy clerks for STDs. We performed a cross-sectional study. Trained medical students visited 62 pharmacies in the city of Porto Alegre during March 2002. These were randomly chosen from a list of 863 registered pharmacies. The students presented to the pharmacy complaining of dysuria and urethral discharge. After obtaining a prescription, or not, they asked for additional instructions to be followed. Immediately after leaving the premises, the instructions were anonymously recorded. Of the 62 pharmacies visited, a clerk in 56 (90.3%, 95% confidence interval [CI]: 80.1%-96.4%) provided a prescription. Most frequently prescribed drugs were ampicillin with probenecide (29/51.8%) and rosoxacin (11/19.6%). Ministry of Health-recommended treatment was not suggested by any of the clerks. Forty-six additional recommendations were given. The use of condoms was the most frequent additional advice (42/46). Prescribing by pharmacy clerks is very prevalent in Porto Alegre. This may represent a lost opportunity for more comprehensive prevention effort (counselling, partner management, and diagnosing other STDs). Additionally, the most frequently prescribed drugs are not recommended by international or national health authorities for treatment of STDs, and none of these drugs covers chlamydia. We conclude that pharmacy clerks are a potentially important source of STD treatment and control but that their practices are in need of vast improvement. PMID- 15117505 TI - Transfusion-associated HIV infection in Mexico related to paid blood donors; HIV epidemic. AB - The objective of the study was to describe the clinical, epidemiological profile and conditional incubation period in a group of transfusion-associated HIV infected (TAHI) patients seen in five national tertiary care centres in Mexico from 1983 to April 1998. Date of transfusions, AIDS diagnoses, opportunistic infections and malignancies were collected. The incubation period was estimated through a non-parametric conditional analysis. One hundred and fifty-seven TAHI cases were analysed. The frequency of TAHI by year of transfusion was: 0.6% in 1980 and 1981, 4.5% in 1984, 22.4% in 1985, 54.5% in 1986, 10.3% in 1987, 0.6% in 1988, 1.9% in 1989 and 1990, 1.3% in 1993 and 0.6% in 1994 and 1996. The median incubation period was 4.3 years. A well-defined epidemic period of HIV-infection among blood-recipients was identified that coincided with the HIV-epidemic among paid donors. TAHI patients in Mexico developed AIDS in a shorter time than that described for other populations. PMID- 15117506 TI - Drug use and sexual risk behaviours among female Russian IDUs who exchange sex for money or drugs. AB - Countries of the former Soviet Union are experiencing the steepest increases in annual HIV incidence in the world. Over 80% of registered HIV cases in Russia have occurred among intravenous drug users (IDUs), but current conditions set the stage for a heterosexually-transmitted epidemic. IDUs who also trade sex for money or drugs may serve as a conduit, or 'bridge' group, through which HIV could make inroads into the general Russian population. The present study examined the prevalence of sex trading among female Russian IDUs, and further examined drug use, sexual behaviour, and perceived vulnerability in this group. Female IDUs (n=100) in St Petersburg, Russia participated; 37% reported a history of sex trading. This group reported a mean of 49.5 male sexual partners in the previous month and an average of 15.4 unprotected vaginal intercourse acts in the previous 30 days. A significant minority (44%) also reported sharing injection equipment with others. Mathematical models to calculate risk estimates for HIV seroconversion indicated that participants were at significant risk of contracting HIV and infecting sexual partners. Despite significant rates of risk behaviours, most participants perceived themselves to be at little risk of contracting HIV. Effective HIV prevention programmes targeted at this group are urgently needed and are likely to be a cost-effective step in curtailing the spread of HIV in the region. PMID- 15117508 TI - Audit of the management of early syphilis at North Manchester General Hospital. AB - The city of Manchester has seen a sustained increase in reported cases of early (infectious) syphilis since the late 1990s. We audited the management of patients presenting with early syphilis to North Manchester General Hospital, with reference to the UK national guidelines. Between January 1999 and December 2001 72 cases of early syphilis were identified. Most (90%) occurred in men who have sex with men, 50% of whom were HIV-positive. Serology and polymerase chain reaction testing of lesions were useful diagnostic tests. Treatment regimens followed the national guidelines in 63% of cases, with adherence to the guidelines improving as the outbreak continued. The majority of patients were treated with intramuscular penicillin (78%), with only three discontinuing this treatment due to side effects. Only 4% of sexual contacts identified were traced and screened, an indication of high levels of both anonymous sex and partner change in this group. PMID- 15117507 TI - The association of neoplasms and HIV infection in the correctional setting. AB - HIV-associated immunosuppression has been linked to an increased risk of a number of cancers, including Kaposi sarcoma (KS), non-Hodgkin's lymphoma (NHL), and invasive cervical cancer. Because prison inmates constitute one of the highest HIV/AIDS prevalent populations in the US, understanding the link between HIV infection and cancer in the correctional setting holds particular public health relevance. The study population consisted of 336,668 Texas Department of Criminal Justice inmates who were incarcerated, for any duration, between 1 January 1999 and 31 December 2001. Inmates diagnosed with HIV infection exhibited elevated rates of KS, NHL, anal cancer, and Hodgkin's disease, after adjusting for age and race. The elevated rates of cancer among HIV-infected individuals, particularly prison inmates, may be mediated, in part, by high-risk behaviours. HIV-associated risk behaviours, including unsafe sexual practices, injection drug use, and prostitution may be associated with cancer-related risk behaviours, such as smoking, excessive alcohol consumption, and poor diet. It will be important for future investigators to examine the association between HIV infection and cancer risk with sufficiently large study cohorts and appropriate longitudinal designs. PMID- 15117510 TI - Promotion and support of optimal feeding practices for infants and young children: a global challenge, a global responsibility. PMID- 15117511 TI - Efforts to promote breastfeeding in the United States: development of a national breastfeeding awareness campaign. PMID- 15117512 TI - Response to "Breastfeeding and human rights" (J Hum Lact. 2003; 19:357-361). PMID- 15117513 TI - Associations between human milk SIgA and maternal immune, infectious, endocrine, and stress variables. AB - Fifty breastfeeding mothers were studied at 4, 5, or 6 weeks postpartum. Morning hind milk, serum samples, and stress and infection data were collected to examine whether milk secretory immunoglobulin A (SIgA) is related to demographic, stress, immune, and infection factors. The authors investigated relationships between milk SIgA and maternal stress, mood, symptoms of infection, serum proinflammatory and proimmune cytokines, and cortisol. Older women had lower SIgA, and smokers and lower-income women had higher SIgA. There was a relationship between postpartum infection symptoms and SIgA concentration. The Profile of Mood States anger score was correlated with higher SIgA and perceived stress with lower SIgA. Positive life events were correlated with higher SIgA. In regression analysis, age, postpartum infections, and serum interferon (IFN)-gamma were significant predictors of milk SIgA. The data suggest that milk SIgA is primarily affected by maternal immune/infection status, but several other stress and mood variables may potentially influence the concentration of milk SIgA. PMID- 15117515 TI - Breast milk expression in the workplace: a look at frequency and time. AB - The objective of this article is to study a barrier for breastfeeding women working full-time outside the home: breast milk expression in the workplace. Data are from a large corporation that provides employee benefits. Mothers express breast milk about twice a day when infants are 4 months old (x = 2.2 +/- 0.8) and 6 months old (x = 1.9 +/- 0.6), with a significant decline in frequency (P <.05) comparing the 2 age groups. Most mothers spend 1 hour or less expressing breast milk when infants are 3 (82%) or 6 months old (96%), with a significant difference (P <.05) between the 2 age groups. Mothers of younger infants were no more likely to work fewer days per week than were mothers of older infants. Most women can express breast milk for 3- and 6-month-old infants in less than an hour, distributed in about 2 separate portions, in an employment environment supportive of breastfeeding. PMID- 15117516 TI - Breastfeeding practices in Colorado businesses. AB - The Colorado Worksite Infant Feeding Survey was used to examine breastfeeding support policies and practices within Colorado businesses. Only 28.2% of survey respondents reported that their organization provided breastfeeding support services. However, the majority of businesses did offer benefits and services conducive to breastfeeding. There was a low frequency of positive responses related to incentives for increasing the provision of breastfeeding support services. The incentives receiving the highest frequency of positive responses were those related to employer benefits and needs, such as provision of information on successful breastfeeding support programs within similar organizations; provision of information on the benefits of breastfeeding to employers, including cost savings; and availability of tax credits for providing breastfeeding support services. Multiple approaches are indicated for increasing work-site breastfeeding support and for removing barriers to the achievement of Healthy People 2010 breastfeeding objectives for employed mothers. PMID- 15117518 TI - Factors associated with the initiation and duration of breastfeeding by Chinese mothers in Perth, Western Australia. AB - To identify determinants of the initiation and duration of breastfeeding by Chinese Australian mothers, a cross-sectional survey of 506 Mandarin-speaking women in Perth, Western Australia, was conducted. Doctors' support of breastfeeding was positively associated with the initiation of breastfeeding both in the mothers' home countries (odds ratio [OR], 9.94; 95% confidence interval [CI], 3.17-31.18) and in Australia (OR, 16.78; 95% CI, 7.12-39.55) and with duration. Mother's level of education was positively associated with the initiation of breastfeeding in the mother's home country (OR, 3.04; 95% CI, 1.36 6.80) and positively associated with the duration of breastfeeding both in the mother's home country and Australia. Father's preference for breastfeeding was positively associated with the initiation of breastfeeding of the mother giving birth in Australia (OR, 4.96; 95% CI, 1.93-12.66). Health professionals can provide invaluable support for mothers initiating and continuing breastfeeding in this population. Prenatal education also needs to emphasize the ways in which fathers can support and contribute to breastfeeding. PMID- 15117517 TI - Mothers performing in-home measurement of milk intake during breastfeeding of their preterm infants: maternal reactions and feeding outcomes. AB - This study compares feeding outcomes and perceptions of mothers using in-home test weights and those who did not use test weights to manage breastfeeding of their preterm infants during the first month after hospital discharge. There were no significant differences in daily weight gain between the 2 groups during the study period. Maternal concerns cited in both groups were similar, namely, knowing how much milk infant is taking, infant gaining adequate weight, and infant getting enough milk. All women in the experimental group and two-thirds in the control group reported that in-home measurement of milk intake by test weighing had been or would have been helpful. This prospective randomized study demonstrated that mothers of premature infants who performed in-home test weighing procedures found the technique to be helpful and experienced no increased stress or lower achievement of breastfeeding goals when compared to mothers not performing test weighs. PMID- 15117519 TI - Family characteristics associated with duration of breastfeeding during early infancy among primiparas. AB - This investigation examined the association of relationship characteristics and parental gender roles with duration of breastfeeding among primiparous women during the early postpartum period. An ethnically diverse group of urban middle class women (n = 115) who initiated breastfeeding were individually surveyed by telephone along with their male partners. Results of the Cox regression analyses confirmed earlier findings reported in the literature. Reporting higher relationship distress was predictive of early breastfeeding cessation at a marginal level of significance. However, levels of maternal responsibility for household tasks and infant care were highly significant predictors of breastfeeding cessation but in opposite directions. These results suggest that the effects of paternal gender roles on breastfeeding success may vary depending on the area of involvement. Findings have implications for further research examining breastfeeding duration, gender roles, and relationship quality and for clinical practice that facilitates adaptive roles and relationships among new parents. PMID- 15117520 TI - Breastfeeding information in pediatric textbooks needs improvement. AB - The objective of this study is to determine if breastfeeding information in general pediatric textbooks used in the United States is accurate and up-to-date. Seven pediatric textbooks published between 1999 and 2002 were reviewed. Three reviewers examined breastfeeding content in each text independently. Using a standardized scoring sheet, each book was evaluated for inclusion of 15 basic breastfeeding criteria. Of the 15 criteria scored, the mean number present in each textbook was 11. The mean number of criteria that were correct was 7.6 (3.4 for incorrect or inconsistent criteria). The mean number of criteria omitted was 4. For each of the texts, the number of correct responses divided by the number of criteria present was 7/11 (64%), 11/14 (79%), 5/11 (45%), 9/11 (82%), 11/13 (85%), 8/9 (89%), and 2/8 (25%). Thus, breastfeeding information in these texts, when not omitted, is highly variable and at times inaccurate and inconsistent. PMID- 15117521 TI - Treating sore, possibly infected nipples. PMID- 15117522 TI - Treating postpartum breast edema with areolar compression. AB - This case study describes an occurrence of gross edema in the breast and areolar tissue of a mother in the first 2 days postpartum that interfered with the early initiation of breastfeeding. The mother developed severe generalized fluid retention during labor and early postpartum. Her breasts were naturally large. The edema in her breasts made the areola and nipple tissue firm and nonpliable. The mother successfully latched her newborn onto her breast after being shown areolar compression (AC), a technique developed and named by the authors. AC reduces nipple and areola edema by using gentle positive pressure on the areola. The baby continued to successfully latch onto the breast after AC was used and taught to the mother. PMID- 15117523 TI - Reverse pressure softening: a simple tool to prepare areola for easier latching during engorgement. AB - Successful breastfeeding requires efficient milk transfer through the nipple areolar complex, which includes subareolar tissue. Subareolar tissue resistance increases during engorgement, when expanded circulation and excess interstitial fluid compete for space with increasing milk volumes. Physiologic and iatrogenic events often combine to produce distortion of breast anatomy. Resulting latch difficulty, delayed milk ejection reflex, poor milk transfer, pain, and nipple damage discourage many mothers. The rationale and technique for a simple intervention developed in practice are described: reverse pressure softening (RPS) before latching significantly reduces resistance of subareolar tissue, temporarily freeing it to interact more efficiently with the baby's mouth. RPS also triggers the milk ejection reflex promptly. The health care provider can perform RPS or teach the mother and her significant others, even by telephone. PMID- 15117524 TI - ILCA's inside track. Dealing with mastitis. PMID- 15117531 TI - Recent references. PMID- 15117533 TI - Can videogames be good for your health? PMID- 15117534 TI - Why did he die? The attributions of cause of death among women widowed in later life. AB - The study aimed to examine the causal attributions of death made spontaneously by older widowed women to explain the deaths of their husbands. The data presented are from two qualitative interview studies using the same methodology. There were 65 widows aged between 55 and 93 years old who had been widowed between 0.25 and 60 years. Data were analysed using a grounded theory method and content analysis. Nearly half of the women gave causal explanations for why their husbands died. These explanations can be best understood in terms of lay illness attributions. These results indicate that a broad analysis of cause of death attributions is important in understanding the bereavement experience. The function of attributions in post-bereavement experiences is also examined. PMID- 15117535 TI - Prospective research in health service settings: health psychology, science and the 'Hawthorne' effect. AB - Health service providers sometimes express concern about the impact of prospective survey research upon patient behaviour. To date, there is little available evidence from which to estimate the likelihood of any 'Hawthorne' effect on patient behaviour in health service settings. We analysed data from one of our own surveys to investigate whether inviting people to participate in research had any impact on their subsequent uptake of a screening service. Findings showed that people sent a questionnaire were slightly faster to take up screening than those not sent a questionnaire. We obtained no significant difference in absolute service uptake rate at six months. PMID- 15117536 TI - 'Does my bump look big in this?' The meaning of bodily changes for first-time mothers-to-be. AB - Research on the impact of bodily changes during the transition to motherhood is contradictory. The aim of the study reported here was to provide more useful insights by employing an inductive qualitative approach. Interviews with six women in the latter stages of pregnancy were analysed drawing upon aspects of interpretative phenomenological analysis and Foucauldian discourse analysis. These analyses suggest generally negative consequence, and discursive constructions that have a greater potential to be limiting than empowering. The impact of gender ideologies on women's ways of being while pregnant is highlighted, as is the importance of developing alternative representations of the female, and the pregnant body, which do not pathologize women. PMID- 15117537 TI - The prediction of health-related behaviour in elementary school children. AB - This study investigates the importance of psychological concepts with regard to health-related behaviour in elementary school children based on self-report data of 802 second- and fourth-graders. The results show significant relations of self efficacy and the choice of appropriate coping strategies to positive health related behaviours. The amount of stress experiences (measured by the Youth Self Report) is associated to health-risky behaviours for the total sample and for the gender subgroups, while the other predictor variables show inconsistent relations for different subgroups. The significance of the results for health-promotion during early developmental stages is discussed. PMID- 15117538 TI - HIV/AIDS among adolescents in Eastern Europe: knowledge of HIV/AIDS, social representations of risk and sexual activity among school children and homeless adolescents in Russia, Georgia and the Ukraine. AB - The two studies reported here focus on knowledge and representations of HIV/AIDS (study 1) plus sexual behaviour and hedonistic values (study 2) among 14-17-year old school children and similar aged shelter children. Results indicate that shelter children are more sexually active, less knowledgeable about means of HIV transmission and are more likely to hold stereotyped representations of those most at risk of infection. Russian respondents were the most sexually active, a finding which could at least be partly explained by their higher levels of hedonistic values. These findings are discussed in the context of a climate of continuing social change in this region. PMID- 15117539 TI - Women's experiences of appearance concern and body control across the lifespan: challenging accepted wisdom. AB - This study adopted a lifespan approach to women's experiences of appearance concern and body control. Thirty-two women (aged 16 to 77) were interviewed about their exercise and food regulation. Results of the grounded theory analysis challenge social constructions of appearance concern as associated principally with the reproductive years, and of the body as malleable, and highlight the complexity of the relationship between appearance concern and body control. Despite frequent persistence of (or increase in) appearance concern beyond young adulthood, 'healthier' responses to appearance concern occurred due to changing priorities and increasing awareness. Findings highlight the utility of an inclusive and qualitative approach, and the absence of simple and sovereign factors determining an individual's levels of appearance concern or body control. PMID- 15117540 TI - Men and chronic illness: a qualitative study of LUTS. AB - In order to determine how men react to and manage chronic conditions 16 men with Lower Urinary Tract Symptoms (LUTS) were interviewed to examine the relationship between detecting symptoms and seeking medical help, perceptions of symptoms and the impact of symptoms on daily life. Transcripts were analysed employing IPA and four major themes identified: Uncertainty/Trying to understand, Implications, Ways of coping and Self-concept/Identity. Uncertainty about the nature and origins of their condition and its gradual onset contributed to the delay in seeking medical advice. Men adopted a range of coping strategies to manage their symptoms. Implications for primary health care consultations are considered. PMID- 15117541 TI - Psychosocial factors in medical and psychological treatment avoidance: the role of the doctor-patient relationship. AB - A community sample of 1106 adults was examined to assess the impact of the doctor patient relationship on participants' avoidance of treatment for a recognized medical or psychological problem. Of five aspects of participants' previous experience with their physicians, all but waiting time predicted participants' self-reported treatment avoidance. In two logistic regression models participants who felt their physicians listened more to their concerns were less likely to avoid treatment for both medical and psychological problems during the previous 12 months. These findings suggest that patients' perceptions of how they are treated by physicians may help explain why many people delay or avoid healthcare treatment, even when faced with a significant health problem. PMID- 15117542 TI - Faith-based and secular pathways to hope and optimism subconstructs in middle aged and older cardiac patients. AB - This study was designed to fill gaps in the new field of positive psychology. Using data from two sequential interviews, this study examined the effect of faith-based and secular pathways to hope and optimism among 226 middle-aged and older patients facing a major medical crisis-cardiac surgery. Structural equation modeling demonstrated that religious faith factors contributed to the agency component of hope and dispositional optimism indirectly through the use of prayer as a coping strategy. Other sociodemographically resourcable factors affected both the agency and pathway components of hope as well as dispositional optimism and dispositional pessimism directly or indirectly through their effects on emotional distress. PMID- 15117543 TI - An assessment of health needs of chronic low back pain patients from general practice. AB - The perceived health needs of 50 chronic low back pain patients were explored within an interview procedure. Patients described symptom and treatment histories, the problems they encountered living with back pain and their severity. Problems as goals for recovery were prioritized. The last two tasks were aided by the use of a Problem Board. Interviews were analysed both quantitatively and qualitatively. Results indicated that the use of this patient centred procedure could be helpful in outlining individual concerns, translatable into recovery goals for intervention, regardless of concurrent medical treatment. Implications of this approach for individual needs assessment in chronic conditions and for planning service development are discussed. PMID- 15117545 TI - Immunomodulating activities of polysaccharides isolated from Panax ginseng. AB - Panax ginseng C.A. Meyer has been traditionally used for the prevention and treatment of various chronic diseases and infections. Ginseng marc is a fibrous and insoluble by-product remaining after the extraction process of ginseng. In this research an extrusion process was employed to disintegrate the insoluble ginseng marc structure, and water-soluble ginseng marc polysaccharide (GMP) was isolated. GMP was examined for immunomodulatory effects in murine peritoneal macrophages. GMP significantly increased the lysosomal phosphatase activity and the phagocytic index of peritoneal macrophages (P<.05). The peritoneal macrophages treated with GMP also produced significantly more H(2)O(2) and nitrite than the control without GMP treatment (P<.05). In addition, GMP (100 microg/mL) significantly increased the cell viability of peritoneal macrophages (P<.05). These results suggest that GMP is an effective nonspecific immunomodulatory agent, and its immunostimulating effects may be due to its ability to stimulate the production of reactive oxygen intermediates. PMID- 15117546 TI - Effect of cacao liquor extract on tumor marker enzymes during chemical hepatocarcinogenesis in rats. AB - This study investigated the effect of cacao liquor extract (CLE) on tumor marker enzymes--alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT), glutathione-S-transferase (GST), and glutathione reductase (GR) activities--in plasma and/or liver of hepatocarcinogenic rats, which were induced with diethylnitrosamine and 2-acetylaminofluorene. Twenty-nine male Sprague-Dawley rats (weighing 150-330 g) were divided into four groups (n = 6-8): normal control group (N), normal group + CLE (NE), cancer group (C), and cancer group + CLE (CE). Analysis of variance showed significant differences (P<.05) in the specific activities of ALP, GGT, and GST between the C and N groups. However, GR activity for the C group was not significantly different compared with the N group. In the CE group, the specific activities of ALP, GGT, GST, and GR were significantly lower (P<.05) compared with the C group. The findings showed that CLE could lower the activity of tumor marker enzymes of rats during hepatocarcinogenesis. Based on the results obtained, polyphenol compounds present in the cacao liquor, extracted by using ethanol, have the potential in decreasing the severity of hepatocarcinogenesis. PMID- 15117547 TI - Differentiation-promoting activity of pomegranate (Punica granatum) fruit extracts in HL-60 human promyelocytic leukemia cells. AB - Differentiation refers to the ability of cancer cells to revert to their normal counterparts, and its induction represents an important noncytotoxic therapy for leukemia, and also breast, prostate, and other solid malignancies. Flavonoids are a group of differentiation-inducing chemicals with a potentially lower toxicology profile than retinoids. Flavonoid-rich polyphenol fractions from the pomegranate (Punica granatum) fruit exert anti-proliferative, anti-invasive, anti-eicosanoid, and pro-apoptotic actions in breast and prostate cancer cells and anti-angiogenic activities in vitro and in vivo. Here we tested flavonoid-rich fractions from fresh (J) and fermented (W) pomegranate juice and from an aqueous extraction of pomegranate pericarps (P) as potential differentiation-promoting agents of human HL-60 promyelocytic leukemia cells. Four assays were used to assess differentiation: nitro blue tetrazolium reducing activity, nonspecific esterase activity, specific esterase activity, and phagocytic activity. In addition, the effect of these extracts on HL-60 proliferation was evaluated. Extracts W and P were strong promoters of differentiation in all settings, with extract J showing only a relatively mild differentiation-promoting effect. The extracts had proportional inhibitory effects on HL-60 cell proliferation. The results highlight an important, previously unknown, mechanism of the cancer preventive and suppressive potential of pomegranate fermented juice and pericarp extracts. PMID- 15117548 TI - Effects of germinated brown rice extracts with enhanced levels of GABA on cancer cell proliferation and apoptosis. AB - In the present work we investigated the effects of brown rice extracts on proliferation and apoptosis of cancer cells. Brown rice extracts were prepared using nongerminated brown rice versus germinated brown rices. Mouse leukemia L1210 cells, human acute lymphoblastic leukemia Molt4 cells, and human cervical cancer HeLa cells were treated with either nongerminated brown rice extract (N ex), water-germinated extract (W ex), chitosan-germinated extract (C ex), glutamic acid-germinated brown rice extract (G ex), or chitosan/glutamic acid germinated brown rice extract (CG ex). The concentrations of gamma-aminobutyric acid (GABA) in the G ex and CG ex were three and 3.3 times higher than the GABA concentration in the N ex, respectively. The G ex and CG ex retarded significantly the proliferation rates of L1210 and Molt4 cells, and the highest retardation rate was with CG ex. In addition, the G ex and CG ex enhanced significantly apoptosis of the cultured L1210 cells, but no significant apoptosis was seen with the other extracts, which have lower concentrations of GABA than G ex and CG ex. These results show that brown rice extracts with enhanced levels of GABA have an inhibitory action on leukemia cell proliferation and have a stimulatory action on the cancer cell apoptosis. PMID- 15117549 TI - Ethanol impairs insulin's actions through phosphatidylinositol 3-kinase. AB - Insulin plays an important role in cell metabolism and proliferation. In the present study, we examined the effect of ethanol on insulin actions such as glucose uptake, DNA synthesis, and c-Jun gene expression. Acute treatment with ethanol (200 mM) for 60 minutes inhibited insulin-stimulated 2-deoxyglucose uptake by 50% in 3T3-L1 adipocytes. Insulin-induced DNA synthesis and c-Jun protein expression were also reduced by ethanol treatment in Rat-1 fibroblasts overexpressing normal human insulin receptor. Ethanol has no effect on tyrosine phosphorylation of the insulin receptor and insulin receptor substrate (IRS)-1. However, association of the insulin receptor and IRS-1 with the Src homology 2 domain of the p85 subunit of phosphatidylinositol 3-kinase (PI3-kinase) was reduced by ethanol. Pretreatment with the antidiabetic drug troglitazone, an insulin-sensitizer, reversed ethanol's inhibition. These results suggest that ethanol specifically inhibits the association of the insulin receptor and IRS-1 with the p85 subunit of PI3-kinase, which is required for increased glucose uptake, DNA synthesis, and c-Jun expression by insulin. PMID- 15117550 TI - Effect of kaikasaponin III obtained from Pueraria thunbergiana flowers on serum and hepatic lipid peroxides and tissue factor activity in the streptozotocin induced diabetic rat. AB - We investigated the effect of kaikasaponin III (KS-III) on Phase I and II enzymes and tissue factor (TF) activity to elucidate the pharmacological actions of this immunosuppressive saponin in the diabetic rat. This compound was obtained from the flower of Pueraria thunbergiana (Leguminosae) by chromatographic isolation. This crude drug (Puerariae Flos) has been used as a therapeutic agent for diabetes mellitus in traditional Korean medicine. KS-III prolonged the bleeding time and plasma clotting time in streptozotocin (STZ)-treated rats and increased the TF activity, suggesting that this compound has anti-thrombosis activity in STZ-induced rats. It also inhibited the formation of malondialdehyde (MDA) and hydroxy radicals in serum and liver, but promoted superoxide dismutase (SOD) activity. Low MDA concentrations and low xanthine oxidase and aldehyde oxidase activities were observed in the KS-III-treated rats, suggesting that such Phase I enzyme activities are the major source of lipid peroxidation. However, KS-III increased Phase II enzyme activities such as SOD, glutathione peroxidase, and catalase, suggesting the activation of free radical-scavenging enzymes. These results suggest that KS-III may exhibit its hypoglycemic and hypolipidemic effects by up-regulating or down-regulating antioxidant mechanisms via the changes in Phase I and II enzyme activities. PMID- 15117551 TI - The depletion of sodium nitrite by lactic acid bacteria isolated from kimchi. AB - Nitrites, whether added or naturally occurring in foods, are potential carcinogens, and controlling their concentrations is important for maintaining a safe food supply. In this study we investigated the depletion of sodium nitrite (150 microg/mL) during the fermentation in Lactobacilli MRS broth at 5, 10, 15, 20, 25, 30, and 36 degrees C by lactic acid bacteria (LAB-A, -B, -C, and -D) isolated from kimchi and Leuconostoc mesenteroides strain KCTC3100. The four species of lactic acid bacteria isolated from kimchi were identified as L. mesenteroides, and all produced depletion of less than 20% of sodium nitrite after 10 days of incubation at 5 degrees C. There was less than 40% depletion after 9 days at 10 degrees C, 86.4-92.8% after 7 days at 15 degrees C, 81.4-87.8% after 4 days and more than 90.0% after 5 days at 20 degrees C, 76.3-85.7% after 3 days and more than 90.0% after 5 days at 25 degrees C, and more than 90.0% after 2 days at 30 and 36 degrees C. The depletion by LAB isolates was similar or higher than that by L. mesenteroides strain KCTC3100, and in particular, the LAB D strain showed the highest depletion effect of all the strains tested, up to 15 degrees C. From these results, the strains isolated from kimchi were very effective for the depletion of sodium nitrite at high temperature, and all sodium nitrite was depleted at the initial period of incubation (1-2 days) at 30 and 36 degrees C. But as the temperature was lowered, the depletion effect of sodium nitrite was decreased in all the strains tested from kimchi. This illustrates that the depletion of nitrite by each strain is subject to the influence of temperatures. PMID- 15117552 TI - Effects of tocopherols and tocotrienols on body composition and bone calcium content in adrenalectomized rats replaced with dexamethasone. AB - Long-term glucocorticoid treatment is associated with severe side effects, such as obesity and osteoporosis. A palm oil-derived vitamin E mixture had been shown previously to be protective against osteoporosis in rats given 120 microg/kg dexamethasone daily for 12 weeks. In this study we determined the effects of two isomers of vitamin E (i.e., palm oil-derived gamma-tocotrienol and the commercially available alpha-tocopherol, 60 mg/kg of body weight/day) on body composition and bone calcium content in adrenalectomized rats replaced with two doses of dexamethasone, 120 microg/kg and 240 microg/kg daily. Treatment period was 8 weeks. gamma-Tocotrienol (60 mg/kg of body weight/day) was found to reduce body fat mass and increase the fourth lumbar vertebra bone calcium content in these rats, while alpha-tocopherol (60 mg/kg of body weight/day) was ineffective. Therefore, in conclusion, palm oil-derived gamma-tocotrienol has the potential to be utilized as a prophylactic agent in prevention of the side effects of long term glucocorticoid use. PMID- 15117553 TI - Inhibitory effects of polyphenols in leaves of Artemisia princeps PAMP on protein fragmentation by Cu(II)-H2O2 in vitro. AB - The leaves of Artemisia princeps PAMP have traditionally been used as teas and foods in Japan. Polyphenols in Artemisia plants have been shown to have inhibitory effects against biological damages. The inhibitory effects of polyphenols in the leaves of A. princeps PAMP were investigated on protein fragmentation induced by Cu(II)-H(2)O(2) in vitro. The total polyphenol content in the leaves of A. princeps PAMP was 4.58%. The condensed tannin content was 0.62% by vanillin assay and 0.14% by proanthrocyanidin assay. The polyphenols in the leaves of A. princeps PAMP inhibited bovine albumin fragmentation by Cu(II) H(2)O(2). The effects of polyphenols in the leaves of A. princeps PAMP were similar to those of tannic acid, studied as a related polyphenol. These results demonstrated that the leaves of A. princeps PAMP have inhibitory effects on protein fragmentation damage. PMID- 15117554 TI - Protective effects of a mixture of dietary agents against 7,12 dimethylbenz[a]anthracene-induced genotoxicity and oxidative stress in mice. AB - We investigated the effects of pretreatment with tomato, garlic, and turmeric, alone and in combination, against 7,12-dimethylbenz[a]anthracene (DMBA)-induced genetic damage and oxidative stress in male Swiss mice. Measurement of the incidence of bone marrow micronuclei as well as the extent of lipid peroxidation and the status of the antioxidants reduced glutathione, glutathione peroxidase, and glutathione-S-transferase in the liver and erythrocytes were used as biomarkers of chemoprotection. In DMBA-treated animals, increased frequency of bone marrow micronuclei was accompanied by enhanced lipid peroxidation and antioxidant depletion. Pretreatment with tomato, garlic, and turmeric alone and a combination of these agents significantly reduced the frequencies of DMBA-induced bone marrow micronuclei as well as the extent of lipid peroxidation. These changes may be mediated by the antioxidant-enhancing effects of the dietary agents. The results of the present study suggest that a diet containing even low levels of different naturally occurring compounds is effective in exerting antigenotoxic effects by inhibiting DMBA-induced oxidative stress. PMID- 15117555 TI - Hypoglycemic effect of Aloe vera gel on streptozotocin-induced diabetes in experimental rats. AB - In the present study an attempt has been made to evaluate the presence of hypoglycemic activity in the alcoholic extract of Aloe vera gel. Effects of oral administration of A. vera extract at a concentration of 200 and 300 mg/kg of body weight on (a) normal fasted rats, (b) oral glucose-loaded rats, and (c) streptozotocin-induced diabetic rats have been studied. A. vera extract maintain the glucose homeostasis by controlling the carbohydrate metabolizing enzymes. PMID- 15117556 TI - Saponins from edible legumes: chemistry, processing, and health benefits. AB - Demand for bean products is growing because of the presence of several health promoting components in edible bean products such as saponins. Saponins are naturally occurring compounds that are widely distributed in all cells of legume plants. Saponins, which derive their name from their ability to form stable, soaplike foams in aqueous solutions, constitute a complex and chemically diverse group of compounds. In chemical terms, saponins contain a carbohydrate moiety attached to a triterpenoid or steroids. Saponins are attracting considerable interest as a result of their diverse properties, both deleterious and beneficial. Clinical studies have suggested that these health-promoting components, saponins, affect the immune system in ways that help to protect the human body against cancers, and also lower cholesterol levels. Saponins decrease blood lipids, lower cancer risks, and lower blood glucose response. A high saponin diet can be used in the inhibition of dental caries and platelet aggregation, in the treatment of hypercalciuria in humans, and as an antidote against acute lead poisoning. In epidemiological studies, saponins have been shown to have an inverse relationship with the incidence of renal stones. Thermal processing such as canning is the typical method to process beans. This study reviews the effect of thermal processing on the characteristics and stability of saponins in canned bean products. Saponins are thermal sensitive. During soaking and blanching, portions of saponins are dissolved in water and lost in the soaking, washing, and blanching liquors. An optimum thermal process can increase the stability and maintain the saponins in canned bean products, which is useful for assisting the food industry to improve thermal processing technology and enhance bean product quality. PMID- 15117557 TI - Differential metabolic effects on mitochondria by silica hydride using capillary electrophoresis. AB - Working as an extension of a newly developed method for a capillary electrophoretic analysis of purine nucleotides, nucleosides, bases, and catabolism, an assay of the differential metabolic properties by a novel organosiliceous anionic hydride compound, silica hydride, was evaluated with Chinese hamster ovary mitochondria using a 50-microm poly(acryloylaminopropanol) coated, fused-silica capillary. The results of this organellar differential analysis indicate a correlation of increased redox pair of NADH to NAD(+) ratios by two times and an increase in ATP levels in the assayed mitochondria by six times. Glucose levels in the organelles were half of the original values. This study validates the electrophoretic method utilizing live organelle fractions for differential metabolic analysis and additionally illustrates some of the emerging novel properties of silica hydride. As confirmation of the results obtained in this assay, additional methods of standard protocol were used to monitor the mitochondrial metabolic activity. PMID- 15117558 TI - Peroxynitrite scavenging activity of indole derivatives: interaction of indoles with peroxynitrite. AB - One of the products of nitrogen-derived free radicals, peroxynitrite (ONOO(-)), is formed by the reaction of superoxide anion (O(2)(*-)) with nitric oxide (NO). ONOO(-) can cause damage to proteins and DNA through nitration. In particular, proteins and their constituent amino acids have been proven to be extremely sensitive to ONOO(-). However, the lack of specific endogenous defense enzymes to protect against ONOO(-) has prompted many researchers to search for endogenous scavengers. We previously found 5-hydroxytryptamine (HT), which is an indole derivative (ID), to be an efficient ONOO(-) scavenger. In the present study, the interaction of several other indoles was further investigated: tryptophan (TRP), 5-hydroxyL-tryptophan (HLT), HT, N-acetyl-5-hydroxytryptamine (AHT), 5 methoxyindole-3-acetate (MIA), 5-methoxytryptamine (MT), and melatonin. The ONOO( ) scavenging activity of ID was assayed by measuring the formation of oxidized dihydrorhodamine-123 (DHR-123). The scavenging efficacy was expressed as the IC(50), denoting the concentration of each indole required to cause 50% inhibition of DHR-123 formation. In a separate in vitro study, the protective effect of IDs against ONOO(-)-induced nitration of bovine serum albumin was investigated. Nitration was quantified using an immunoassay with a monoclonal anti-nitrotyrosine antibody, and a horseradish peroxidase-conjugated anti-mouse secondary antibody from sheep. The results revealed that the inhibitory activities of indoles were as follows: HLT, IC(50) = 0.73 microM; HT, IC(50) = 1.03 microM; and AHT, IC(50) = 0.98 microM), showing relatively strong activities against ONOO(-). Interestingly, TRP, MIA, MT, and melatonin were less effective. Regarding the protection of albumin by IDs, the data showed that the formation of ONOO(-) was inhibited in a dose-dependent manner. Further probing of the mode of the interaction of indoles revealed that the hydroxyl groups in IDs are required for the enhanced scavenging action. It was concluded that several indole derivatives with hydroxyl groups are effective scavengers against ONOO(-), and that the scavenging efficacy depends on the presence of hydroxyl groups located within the indole ring structure. PMID- 15117559 TI - An in vitro study on antimicrobial activity of propolis from Mugla province of Turkey. AB - Antibacterial and antifungal activities of acetone and dimethyl sulfoxide (DMSO) extracts of 45 different propolis samples from the Mugla province of Turkey were investigated. Antimicrobial activity of propolis varied depending on propolis sample, dosage of propolis, and the extraction solvents for all test microorganisms. Antimicrobial activity of all propolis samples increased with increasing dosage without reaching a plateau at the highest dosage tested. Except for Brucella melitensis, the DMSO extracts of all propolis samples were more active than the acetone extracts of the same samples. For B. melitensis, the acetone extracts of all propolis samples showed greater activity. The most sensitive microorganism to propolis was Shigella sonnei in the gram-negative group and Streptococcus mutans in the gram-positive group. The least sensitive microorganism was Candida albicans. A control test run with standard antibiotics revealed that propolis samples from the Mugla province of Turkey has a similar or greater inhibitory effect on S. mutans, Salmonella typhi, Pseudomonas aeruginosa, S. sonnei, and C. albicans growth. PMID- 15117560 TI - Effects of different processing methods on induction of quinone reductase by dietary broccoli in rats. AB - Broccoli belongs to a group of cruciferous vegetables characterized by its content of glucosinolates, secondary metabolites that, upon hydrolysis, release bioactive isothiocyanates (ITCs). Sulforaphane, the major ITC from broccoli, is believed to protect the body from cancer by induction of detoxification enzymes such as quinone reductase (QR). Sulforaphane provides powerful protection against carcinogenesis, mutagenesis, and other forms of toxicity by electrophiles and reactive forms of oxygen. The purpose of this study was to investigate the effects of processing methods on the ability of broccoli to induce QR in various rat tissues. Male F344 rats (four per group) received an AIN 76B-40 diet containing either 0% or 20% broccoli processed by different methods (dehydrated, freeze-dried, or freeze-dried and hydrolyzed) for 5 days. Colon tissues of rats receiving dehydrated, freeze-dried, and hydrolyzed broccoli diets showed QR induction of 9.1-, 10.5-, and 6.4-fold, respectively. Induction of QR by dehydrated broccoli in the liver and kidney was significantly less robust than in colon, being 2.3- and 1.6-fold over control, respectively. These results suggest that freeze-drying and dehydration are promising approaches for providing the public with the functional benefits of broccoli consumption. PMID- 15117561 TI - Natural honey lowers plasma glucose, C-reactive protein, homocysteine, and blood lipids in healthy, diabetic, and hyperlipidemic subjects: comparison with dextrose and sucrose. AB - This study included the following experiments: (1) effects of dextrose solution (250 mL of water containing 75 g of dextrose) or honey solution (250 mL of water containing 75 g of natural honey) on plasma glucose level (PGL), plasma insulin, and plasma C-peptide (eight subjects); (2) effects of dextrose, honey, or artificial honey (250 mL of water containing 35 g of dextrose and 40 g of fructose) on cholesterol and triglycerides (TG) (nine subjects); (3) effects of honey solution, administered for 15 days, on PGL, blood lipids, C-reactive protein (CRP), and homocysteine (eight subjects); (4) effects of honey or artificial honey on cholesterol and TG in six patients with hypercholesterolemia and five patients with hypertriglyceridemia; (5) effects of honey for 15 days on blood lipid and CRP in five patients with elevated cholesterol and CRP; (6) effects of 70 g of dextrose or 90 g of honey on PGL in seven patients with type 2 diabetes mellitus; and (7) effects of 30 g of sucrose or 30 g of honey on PGL, plasma insulin, and plasma C-peptide in five diabetic patients. In healthy subjects, dextrose elevated PGL at 1 (53%) and 2 (3%) hours, and decreased PGL after 3 hours (20%). Honey elevated PGL after 1 hour (14%) and decreased it after 3 hours (10%). Elevation of insulin and C-peptide was significantly higher after dextrose than after honey. Dextrose slightly reduced cholesterol and low-density lipoprotein-cholesterol (LDL-C) after 1 hour and significantly after 2 hours, and increased TG after 1, 2, and 3 hours. Artificial honey slightly decreased cholesterol and LDL-C and elevated TG. Honey reduced cholesterol, LDL-C, and TG and slightly elevated high-density lipoprotein-cholesterol (HDL-C). Honey consumed for 15 days decreased cholesterol (7%), LDL-C (1%), TG (2%), CRP (7%), homocysteine (6%), and PGL (6%), and increased HDL-C (2%). In patients with hypertriglyceridemia, artificial honey increased TG, while honey decreased TG. In patients with hyperlipidemia, artificial honey increased LDL-C, while honey decreased LDL-C. Honey decreased cholesterol (8%), LDL-C (11%), and CRP (75%) after 15 days. In diabetic patients, honey compared with dextrose caused a significantly lower rise of PGL. Elevation of PGL was greater after honey than after sucrose at 30 minutes, and was lower after honey than it was after sucrose at 60, 120, and 180 minutes. Honey caused greater elevation of insulin than sucrose did after 30, 120, and 180 minutes. Honey reduces blood lipids, homocysteine, and CRP in normal and hyperlipidemic subjects. Honey compared with dextrose and sucrose caused lower elevation of PGL in diabetics. PMID- 15117562 TI - The potential beneficial effect of glycine on the carbohydrate moieties of glycoproteins in an experimental model of alcohol-induced hepatotoxicity. AB - Glycine is known to have a protective role against alcohol-induced liver damage. The aim of our study was to evaluate the effect of glycine on liver and brain glycoproteins in alcohol-fed rats. Administering ethanol (7.9 g/kg of body of weight) every day to Wistar rats for 60 days resulted in significantly elevated levels of liver and brain hexosamine, fucose, and sialic acid and significantly reduced levels of total hexoses as compared with those of the control rats. Simultaneous glycine supplementation (0.6 g/kg of body weight) during the last 30 days of the experiment to rats given alcohol normalized the levels of hexosamine, fucose, and sialic acid and elevated the levels of total hexoses in the liver and brain significantly as compared with unsupplemented alcohol-treated rats. Microscopic examination of alcohol-fed rat liver showed inflammatory cell infiltrates and fatty changes, which were reversed on treatment with glycine. Similarly, alcohol-treated rat brain demonstrated edema, which was markedly reduced on treatment with glycine. Thus glycine administration plays a significant role in reducing the toxicity of ethanol. PMID- 15117563 TI - Inhibition of Aspergillus parasiticus NRRL 2999 by pollen and propolis extracts. AB - The inhibitory effects on mycelial growth of Aspergillus parasiticus strain NRRL 2998 of pollen and propolis extracts from different regions of Turkey were investigated in culture media. The least active concentration towards the tested fungus was 2% of both extracts. But, the inhibitory effects of all propolis extracts on mycelial growth were higher when compared with pollen extracts. While the effect of a 5% level of Antakya propolis extract increased with increasing incubation period, the antifungal effect of pollen extract from the same region on mycelial growth was less than that of the control group. As a result, none of the extracts tested completely inhibited mycelial growth. The highest inhibition rate was established at the 5% level of Taskent and Alanya propolis samples. PMID- 15117564 TI - Binding affinities of ocular hypotensive beta-blockers levobetaxolol, levobunolol, and timolol at endogenous guinea pig beta-adrenoceptors. AB - The current study determined the relative affinities and selectivities of numerous beta-adrenoceptor antagonists at the endogenous beta(1)- and beta(2) adrenoceptors in guinea pig heart and lung, respectively, using [(3)H]-CGP12177. Specific binding of [(3)H]-CGP12177 comprised 80 +/- 0.2% (n = 11) and 94 +/- 0.2% (n = 16) of the total binding in washed heart and lung homogenates, respectively. Concentration-dependent displacement of [(3)H]-CGP12177 binding from beta-adrenoceptors in both preparations was observed with nine different beta-adrenoceptor antagonists. Levobetaxolol, betaxolol, CGP-20712A, levobunolol, and timolol yielded bi-phasic (two-site-fit) competition curves in the heart, while CGP-20712A, ICI-118551 and levobunolol produced bi-phasic curves in the lung preparation. The high-affinity component of [(3)H]-CGP12177 binding in the heart and lung reflected binding to beta(1)-receptors and beta(2)-receptors, respectively. The binding inhibition parameters (IC(50)s) for displacement of [(3)H]-CGP12177 from these predominantly high-affinity sites were: levobetaxolol (24.9 +/- 1.6 nM heart, 4810 +/- 367 nM lung), racemic betaxolol (37.9 +/- 8.7 mM heart; 8840 +/- 424 mM lung), CGP-20712A (4.6 +/- 0.9 nM heart; 171,000 +/- 109,000 nM lung), ICI-118551 (9230 +/- 3240 nM heart; 2.9 +/- 0.6 nM lung), levobunolol (42 +/- 15 nM heart, 0.3 +/- 0.2 nM lung), (l)-timolol (3.1 nM heart, 2.9 +/- 1.5 nM lung), ICI-215001 (5840 +/- 114 nM heart; 26100 +/- 3200 nM lung), BRL-37344 (83,300 +/- 2660 nM heart; 13,200 +/- 1250 lung). These data indicated that while levobetaxolol and betaxolol possessed a 193-233-fold selectivity for beta(1)-receptors, levobunolol exhibited a 140-fold beta(2)-receptor selectivity and (l)-timolol was essentially nonselective. PMID- 15117565 TI - Prostaglandins F2alpha and E2 in aqueous humor of patients with cataract surgery. AB - To understand the role of prostaglandins F(2alpha) and E(2) in aqueous humor under the normal condition, their concentrations were measured by enzyme immunoassay in aqueous fluid obtained from either eye of 60 patients during cataract surgery and correlated with clinical factors as the age and intraocular pressure. The concentrations of prostaglandin F(2alpha) in the aqueous of all patients were below the level of detection. The concentrations of prostaglandin E(2) in the aqueous were below the level of detection in 37 patients while ranged from 9 to 48 pg/mL (median, 31) in 23 patients. The levels of prostaglandin E(2) did not have correlation with the age of the patients or the intraocular pressure of the eyes. In conclusions, the concentrations of prostaglandin E(2) were apparently higher than the concentrations of prostaglandin F(2alpha) in the aqueous. This study could not prove the relationship between prostaglandin levels and the intraocular pressure or the age. PMID- 15117566 TI - Effects of some natural flavonoids on retinal function recovery after ischemic insult in the rat. AB - PURPOSE: There are six natural flavonoids studied recently and their effects on ocular blood flow measured with colored microsphere technique. It was found that three out of six compounds showed strong positive effects in increasing the ocular blood flow. In this study, we tried to find out whether these results can be translated on their effects to improve retinal function recovery after ischemic insult. METHODS: Electroretinography was used to measure the b-wave recovery as an indication of retinal function recovery. RESULTS: Naringenin, hesperetin, and rutin were found to produce marked positive effects on b-wave recovery, whereas naringin, hesperidin, and quercetin showed poor recovery of b wave after ischemic insult of the retina. CONCLUSION: It was found that the compounds that showed strong increase of ocular blood flow also showed marked increase of retinal function recovery, whereas those that showed poor increase of ocular blood flow also showed poor effects on the retinal function recovery. PMID- 15117567 TI - Efficacy of bimatoprost 0.03 percent in untreated glaucoma and ocular hypertension patients: results from a large community-based clinical trial. AB - PURPOSE: To evaluate the intraocular pressure- (IOP-) lowering efficacy of bimatoprost 0.03% (Lumigan, Allergan, Inc.) monotherapy in the treatment of patients with glaucoma or ocular hypertension not currently using ocular hypotensives. METHODS: Open-label, community-based, multicenter evaluation. Patients (n = 6767) who, according to their physicians, required IOP lowering were prescribed bimatoprost for 2 months. Subgroup analyses of the results, stratified by treatment history and use of concomitant medications, were performed. This report focuses on the subgroup of patients that was not being treated with antiglaucoma medications at baseline (n = 1946, 29%). All of these patients were placed on bimatoprost monotherapy. RESULTS: The mean IOP at the untreated baseline was 23.8 mmHg. Bimatoprost provided a mean IOP reduction of 7.5 mmHg (30%, p < 0.001) from baseline after 2 months of monotherapy. Further, bimatoprost allowed patients to achieve low target pressures. For example, 41.5% of patients achieved target IOPs of < or =15 mmHg after 2 months of bimatoprost monotherapy, and 75.8% of patients reached IOPs of < or =18 mmHg. The most commonly reported adverse event was conjunctival hyperemia (7.9%). CONCLUSION: Bimatoprost monotherapy was well tolerated and reduced IOP by an average of 30% in a large population of untreated patients. PMID- 15117568 TI - Sterility of glaucoma medications among chronic users in the community. AB - PURPOSE: To evaluate the sterility of topical glaucoma medications among chronic glaucoma medication users in the community. SETTING: Glaucoma service, Sanz Medical Center, Laniado Hospital, Netanya. Research mode: Cross-sectional laboratory and clinical study. PATIENTS AND METHODS: Chronic glaucoma patients were asked to submit their topical glaucoma preparations to the microbiology laboratory at Laniado hospital. Samples taken from the interior of the bottle and the tip were cultured using MacConkey agar, blood agar, and chocolate agar plates. RESULTS: Sixty-two bottles of topical glaucoma medications used by 27 patients were tested. Bacterial growth was detected in eight (12.9%) preparations, three of which revealed Pseudomonas aeruginosa, three Staphylococcus epidermidis, one Streptococcus viridans, and one Klebsiella. During the study, acute conjunctivitis was found in one patient, possibly due to the use of infected drops. In another case, two identical medications, which had been used simultaneously for more than 6 weeks by the same patient, were found to be infected. CONCLUSIONS: Glaucoma topical preparations are generally found safe in terms of sterility, though bacterial growth may be found in a small percentage. In most cases, the cause of the loss of sterility could not be determined. However, in two preparations, contamination was related to the noncompliance of the patient who continued using the same preparation longer than instructed. It is imperative to increase the awareness of glaucoma patients to the fact that improper use can lead to eye-drop contamination. PMID- 15117569 TI - Mitomycin C alters corneal stromal wound healing and corneal haze in rabbits after argon-fluoride excimer laser photorefractive keratectomy. AB - PURPOSE: To investigate the effects of mitomycin C (MMC) on rabbit cornea wound healing after photorefractive keratectomy (PRK). MATERIALS AND METHODS: Rabbit corneas were stained with dichlorotriazinyl aminofluorescein immediately after PRK. MMC was applied to the right eye and phosphate-buffered salt solution (PBS) to the left. Corneal epithelial wound healing rate and corneal haze were examined. Ultrasound pachymetry was performed. Stromal collagen regeneration was evaluated by fluorescent microscopy. We used terminal deoxyribonucleotidyl transferase-mediated D-uridine 5'-triphosphated-digoxigenin nick-end labeling (TUNEL) assay and transmission electron microscopy (TEM) to evaluate keratocyte apoptosis. RESULTS: In eyes treated with MMC, there was no delay to the healing rate of corneal epithelial wound, and less haze 4 weeks after PRK. Ultrasound pachymetry showed thinner corneal thickness in MMC-treated eyes at week 4. Corneal stromal thickness regression was less in MMC-treated eyes observed by fluorescent microscope at week 4. Keratocyte apoptosis was noted in both MMC- and PBS-treated eyes by TUNEL assay and TEM observation. This study discovered the phenomenon that MMC prolongs keratocyte apoptosis. CONCLUSIONS: Applying MMC after PRK is an effective method to decrease haze formation and corneal stromal thickness regression in rabbit corneas. The effect may be related to MMC prolonging keratocyte apoptosis. PMID- 15117570 TI - A comparative trial of the safety and efficacy of 0.1 percent pemirolast potassium ophthalmic solution dosed twice or four times a day in patients with seasonal allergic conjunctivitis. AB - PURPOSE: To compare efficacy and safety between twice-daily and four-times-daily regimens of pemirolast 0.1% in allergic conjunctivitis patients. METHODS: This investigator-masked trial recruited 169 patients, with a positive skin prick test, +2 itching and hyperemia. Patients were randomized to two times daily (t.i.d.) or four times daily (q.i.d.) treatment during allergy season. Evaluation was at 0, 2 and 4 weeks, with itching and hyperemia at week 4 as the primary endpoints. Analysis used last observation carried forward (LOCF) and analysis of variance (ANOVA) for efficacy, factoring treatment and center variations. The basis of the statistical evaluation was to confirm parity between two treatments, via noninferiority hypothesis testing. A 95% confidence interval (CI) with an upper limit of < or = 0.5 was set to assess non-inferiority or to conclude if schedules were statistically similar. RESULTS: B.i.d. and q.i.d. baselines were similar, respectively, for itching (2.6 and 2.8) and hyperemia (2.3 and 2.2). Week 4 itching was statistically non-inferior between treatments (1 b.i.d. versus 0.8 q.i.d.), with a mean treatment difference of 0.17 (-0.13, 0.47, Delta < or = 0.5). Week 4 hyperemia was comparable (1.2 for b.i.d. versus 1.0 for q.i.d). Week 2 scores and mean change from baseline (weeks 2 and 4), patient diary data, and investigator assessments were comparable. Both regimens were well tolerated with no differences in adverse events were observed. CONCLUSIONS: B.i.d. dosing was statistically non-inferior to q.i.d. dosing with respect to itching and hyperemia. Both regimens were similarly well tolerated in allergic conjunctivitis patients. PMID- 15117571 TI - Effects of hyaluronan on free-radical formation, corneal endothelium damage, and inflammation parameters after phacoemulsification in rabbits. AB - Free-radical formation may play a role in postoperative complications of phacoemulsification (e.g., corneal endothelium damage from mechanical injury). The present experiments were aimed at investigating whether different molecular weight ranges (2000-2600, 2600-3200, or 3200-3800 kDa) of hyaluronan may influence free radical formation, corneal endothelium damage, and inflammation parameters after phacoemulsification in the rabbit eye. The viscoelastic substance was injected in the anterior chamber of rabbits' eyes before phacoemulsification, at a 2.5% concentration. The formation of free radicals was determined by adding luminol to the irrigation media and measuring the chemoluminescence in eyes. The corneal endothelial damage was evaluated by measuring the corneal central thickness by pachimetry. The inflammation parameters were measured by calculation in aqueous humor of peak levels of leukocytes and prostaglandin E(2) (PGE(2)) and evaluation in uveal tissue of myeloperoxidase activity. Hyaluronan decreased by about 58-60% free-radical formation during phacoemulsification, reduced by about 76-80% modifications in mean corneal thickness and by about 54-61% the corneal endothelial cell loss in all molecular weight ranges used. No difference was found among various molecular weight ranges. The highest molecular weight range showed to be more potent than the lowest range for reduced number of inflammation cells and level of PGE(2) in aqueous humor. Thus, hyaluronan reduces free-radical formation, exerts protection on the corneal endothelium and exerts anti-inflammation properties after phacoemulsification in rabbits. The latter effect seems to depend on the molecular weight of the substance. PMID- 15117572 TI - Scleral permeability of a small, single-stranded oligonucleotide. AB - Developing more effective ocular drug delivery systems is essential to improving the treatment of posterior segment eye disease. The large target area provided by the sclera and potentially less vision threatening complications are advantages of transscleral administration compared to more traditional modalities of drug delivery to the posterior segment. We aimed to determine the permeability coefficient for the in vitro diffusion of a small, single-stranded, oligonucleotide across human sclera. Transscleral permeability was measured by placing 100 microL of 2.96 x 10(-4) mol single-stranded, fluorescein-labeled oligonucleotide (MW = 7998.3) on the episcleral surface of sclera mounted in a perfusion chamber. Fractions of choroidal perfusate were collected hourly for 24 hours. The permeability constant or K(trans) for the transscleral diffusion of the naked, single-stranded, fluorescein-labeled oligonucleotide was 7.67 +/- 1.8 x 10(-7) cm/s (mean +/- SEM, N = 7). The permeability constant or K(trans) after intrascleral injection of the same fluorescein-labeled oligonucleotide was 1.32 +/- 0.42 x 10(-7) (mean +/- SEM, N = 4). This analysis demonstrates that diffusion of a naked, 24-base, single-stranded, fluorescein-labeled oligonucleotide can be accomplished by both of the described methods. The ability to deliver single-stranded oligonucleotides across the sclera may prove to be advantageous given the development of several novel therapeutic strategies that use similar molecules. PMID- 15117573 TI - Comparison of liposome-encapsulated acyclovir with acyclovir ointment: ocular pharmacokinetics in rabbits. AB - A positively charged liposomal formulation for topical administration of acyclovir (ACV) was investigated in comparison with a commercial ACV ointment, by determining the pharmacokinetic profile of the drug in the aqueous humor of rabbits after topical administration. The ointment was tested at two different strengths: undiluted (3.0%) and diluted to the same ACV concentration as the liposomal vehicle (0.12%). A liquid formulation containing ACV plus "empty" liposomes and an isotonic aqueous ACV solution were also tested. The applied ACV dose was 0.18 mg, except for the full-strength (3.0%) ointment, in which case it was 1.5 mg. The ACV liposomal dispersion (LIPO-ACV) produced a significantly higher drug concentration profile in the aqueous with respect the three reference formulations containing the same ACV concentration, and showed a 90-minute plateau. The aqueous humor ACV concentration maintained by LIPO-ACV during the plateau was in the upper range of the ID(50)s (0.01 to 0.7 microg/mL) reported for Herpes simplex type 1. In spite of the much higher dose (1.5 versus 0.18 mg), the area under curve (AUC) produced by the full-strength 3.0% ointment was only 1.6 times greater than that corresponding to the liposomal vehicle. In vitro release tests through a cellophane membrane substantiated the concept that positively charged liposomal formulations owe their efficacy to interactions with the positively charged corneal epithelium. PMID- 15117574 TI - Ocular hemorrhage possibly the result of HMG-CoA reductase inhibitors. AB - This retrospective case series describes the association between ocular hemorrhage and 3-hydroxy-3-methylglutaryl conenzyme A (HMG-CoA) reductase inhibitors (statins). The clinical characteristics of 95 case reports submitted to the World Health Organization (WHO), the Food and Drug Administration, and the National Registry of Drug-Induced Ocular Side Effects are summarized with classification of this ocular side effect according to WHO criteria. The average time to onset of ocular hemorrhage was 300 days with 11 positive dechallenge reports and 2 positive rechallenge cases. Some patients also received medications known to increase bleeding times. From the collected data, ocular hemorrhage is "possibly" due to statin therapy. PMID- 15117576 TI - Alarms based on real-time sensor glucose values alert patients to hypo- and hyperglycemia: the guardian continuous monitoring system. AB - The purposes of this study were to demonstrate the accuracy and effectiveness of the Guardian Continuous Monitoring System (Medtronic MiniMed, Northridge, California) and to demonstrate that the application of real-time alarms to continuous monitoring alerts users to hypo and hyperglycemia and reduces excursions in people with diabetes. A total of 71 subjects with type 1 diabetes, mean hemoglobin A1c of 7.6 +/- 1.1%, age 44.0 +/- 11.4 years, and duration of diabetes 23.6 +/- 10.6 years were enrolled in this two-period, randomized, multicenter study. Subjects were randomized into either an Alert group or a Control group. The accuracy of the Guardian was evaluated by treating the study data as a single-sample correlational design. Effectiveness of the Guardian alerts was evaluated by comparing the Alert group with the Control group. The mean (median) absolute relative error between home blood glucose meter readings and sensor values was 21.3% (17.3%), and the Guardian, on average, read 12.8 mg/dL below the concurrent home blood glucose meter readings. The hypoglycemia alert was able to distinguished glucose values < or =70 mg/dL with 67% sensitivity, 90% specificity, and 47% false alerts. The hyperglycemia alert showed a similar ability to detect sensor values > or =250 mg/dL with 63% sensitivity, 97% specificity, and 19% false alerts. The Alert group demonstrated a median decrease in the duration of hypoglycemic excursions (-27.8 min) that was significantly greater than the median decrease in the duration of hypoglycemic excursions in the Control group (-4.5 min) (P = 0.03). A marginally significant increase in the frequency of hyperglycemic excursions (P = 0.07) between Period 1 and Period 2 was accompanied by a decrease of 9.6 min in the duration of hyperglycemic excursions in the Alert group. Glucose measurements differ between blood samples taken from the finger and interstitial fluid, especially when levels are changing rapidly; however, these results demonstrate that the Guardian is reasonably accurate while performing continuous glucose monitoring. The subjects' responses to hypoglycemia alerts resulted in a significant reduction in the duration of hypoglycemic excursions; however, overtreating hypoglycemia may have resulted in a marginally significant increase in the frequency of hyperglycemic excursions. PMID- 15117577 TI - The role of new technology in the early detection of hypoglycemia. PMID- 15117578 TI - Impact of particle size and aerosolization time on the metabolic effect of an inhaled insulin aerosol. AB - The effects were compared of varying aerosol particle size and aerosolization time within each breath on the metabolic effect elicited by inhalation of a liquid insulin aerosol in comparison with that after subcutaneous injection (s.c.) of regular insulin. In this single-center, open-label euglycemic glucose clamp study, 13 healthy non-smoking subjects received five administrations of insulin in randomized order on separate study days, once by s.c. (0.15 U/kg of regular insulin) and four times by inhalation. Subjects inhaled 1.5 U/kg of liquid insulin aerosol administered by the Aerodose Insulin Inhaler (Aerogen Inc., Mountain View, CA) configured to deliver two aerosol particle sizes--fine [F, 4.4 +/- 0.3 microm (mean +/- SD)] or very fine (VF, 3.5 +/- 0.2 microm)--and two aerosolization times (aerosol released for the first 2 or 4 s after the start of each 5-s inhalation). Glucose infusion rate (GIR) values necessary to keep blood glucose concentrations constant at 5.0 mmol/L were determined over a 6-h period following insulin administration. After inhalation of insulin, the onset of action was substantially more rapid on all four inhalation study days than after s.c. insulin, and the time to maximal action [t(GIRmax) (min)] was reached earlier: F/2 s, 127 +/- 54; F/4 s, 128 +/- 55; VF/2 s, 158 +/- 91; VF/4 s, 132 +/ 72; s.c., 175 +/- 69 (P < 0.0001). The longer aerosolization time (4 vs. 2 s) resulted in higher maximal metabolic action [GIR(max) (mg/kg/min), F/4 s 8.1 +/- 3.6, VF/4 s 8.4 +/- 2.7 vs. F/2 s 6.6 +/- 2.4, VF/2 s 7.2 +/- 2.4 (P = 0.01 for 4 s vs. 2 s, grouped data)], total metabolic activity [area under the curve of GIR 0-6 h (g/kg), F/4 s 1.97 +/- 0.92, VF/4 s 2.14 +/- 0.86 vs. F/2 s 1.56 +/- 0.68, VF/2 s 1.78 +/- 0.60 (P = 0.01)], and relative biopotency [F/4 s 10.6 +/- 4.0%, VF/4 s 11.7% +/- 4.1% vs. F/2 s 8.5 +/- 3.2%, VF/2 s 9.7 +/- 2.4% (P = 0.01)]. None of these summary measures was significantly affected by particle size. No drug- or device-related adverse events were observed. This study shows that aerosolization time, but not particle size, in the ranges studied, had an impact on the metabolic effect elicited by inhaled insulin, allowing rational selection of delivery parameters for further clinical testing. Based on the observed biopotency and the rapid onset of action, inhalation of a liquid insulin aerosol generated by the Aerodose Insulin Inhaler shows promise for covering prandial insulin requirements. PMID- 15117579 TI - Cardiac and vascular function in adolescents and young adults with type 1 diabetes. AB - The objective was to prospectively define cardiac and vascular function in adolescents and young adults with type 1 diabetes over a 3-year period. Thirty three subjects with type 1 diabetes were studied in 1998, and 28 returned for follow-up in 2001. There were 28 controls without diabetes. All subjects underwent ultrasonography of the carotid artery to determine distensibility, compliance, and intimal-medial thickness, as well as echocardiographic studies, lipid profile, and measurement of hemoglobin A1c (HbA1c) levels and urinary albumin excretion rate. The diabetes group showed no significant change over the 3-year follow-up in lipid profile, HbA1c level, or albumin excretion rate, but a significant increase in body mass index. The diabetes and control groups were similar in age, lipid profile, and albumin excretion rate. Carotid artery distensibility and compliance in the diabetes group increased significantly from baseline to follow-up but did not differ significantly from controls. The intimal medial thickness of the diabetes group remained unchanged over time but tended to be higher than controls. Echocardiographic studies showed no difference between the diabetes and control groups, but there was a small change in left ventricular posterior diastolic wall diameter in the subjects with diabetes over the 3-year period. There was an overall improvement in carotid function and no change in cardiac measures in subjects with diabetes over the 3 years, such that cardiac and vascular function were similar to those of controls. The reason for the improvement is uncertain, but may relate to the improvement of insulin sensitivity at the end of the pubertal growth period, rather than to changes in metabolic control or lipid profiles. PMID- 15117580 TI - The effect of local subcutaneous delivery of vascular endothelial growth factor on the function of a chronically implanted amperometric glucose sensor. AB - The foreign body capsule that forms around implanted devices such as glucose sensors is hypovascular and has limited permeability to glucose. Such a capsule may function better if well vascularized. We hypothesized that capsular vascularization achieved by local release of vascular endothelial growth factor (VEGF) would lead to enhanced function. Amperometric glucose sensor array disks, each with four indicating electrodes, were implanted into rats. Animals received local subcutaneous infusions of VEGF(165) via osmotic pumps at a location on the sensor face 2 mm from one of the electrodes ("near units"). "Intermediate" electrode units were 15 mm, and "distant" units were 22 mm, from the VEGF source. Every 2 weeks, a glucose infusion was given to assess sensor function by telemetry. Near units demonstrated a lower lag duration (delay after blood glucose) than intermediate and distant units. The mean absolute relative difference for near units was less than for distant units. The percentage of data pairs in the A region of the Clarke error grid of the near sensing units was greater than that of the distant units. Values for the functional measures for saline controls fell between near and distant VEGF values. Glucose sensor function was found to be more favorable in units immediately adjacent to the VEGF infusion port. The most likely cause for this finding is increased neovessel growth in the surrounding foreign body capsule. Slow release of angiogenic growth factors may be a potential method for chronically enhancing the function of a subcutaneously implanted biosensor. PMID- 15117581 TI - Relating glucose clamp profiles to reduction of blood glucose after insulin administration. AB - A model was established allowing prediction of blood glucose response from glucose clamp results performed in healthy volunteers. Data from published studies performed in healthy volunteers were used to establish, test, and validate a model for the evaluation of glucose reductions from glucose clamp results. Studies included those that measured blood glucose and glucodynamic response over time after administration of 0.05 U/kg of regular human insulin (HR) and insulin lispro (LP) with and without the benefit of a glucose clamp procedure. An inhibitory effect E(max) model was used to describe the relationship; the model differed between the HR and LP responses by the intensity of the counterregulatory response as assessed by glucagon measurements. The relationships were used to predict blood glucose responses from a clamp study assessing NPH insulin and HR administrations. Glucose concentrations measured after administration of NPH insulin and HR without a clamp were compared to the model-predicted results to assess the accuracy of the model predictions. The E(max) model successfully correlated the glucose clamp results with the blood glucose depressions in the presence and absence of a counterregulatory response. However, predictions of glucose depression were only accurately modeled in the absence of a counterregulatory glucagon response. The correlations established with a minimal counterregulatory response underscore the value of glucose clamp procedures in defining the time-activity profiles of insulins when the clamp is established at fasting glucose concentrations. PMID- 15117582 TI - Automated insulin infusion trials in the intensive care unit. AB - The objective is to demonstrate the effectiveness of a simple automated insulin infusion for controlling the rise and duration of blood glucose excursion following a glucose challenge in critically ill patients with impaired glucose tolerance. A two-compartment model of the glucose regulatory system was developed for intravenous infusion control design. On two subsequent days a critically ill patient with impaired glucose tolerance was given a 75 g oral glucose tolerance test (OGTT), and the glucose level was measured every 15 min. The first day's data were used to design a heavy-derivative insulin infusion controller for the second day. Ethics approval was granted for this test. Five patients were studied. In four patients, the magnitude and duration of blood glucose excursion were reduced over 50%. Fasting level was reduced 15%, from an average of 7.2 mmol/L to 6.1 mmol/L. The fifth patient's results showed a diminished response due to the antagonistic effects of hydrocortisone on insulin, a data point not provided prior to testing. Modeling to account for this effect yielded better correlation with the test. The automated algorithm provided rapid, effective control of the blood glucose rise in response to an OGTT input. These results highlight the effectiveness of automated infusions for regulating blood glucose rise and excursions, and the potential of this approach for non-hospitalized individuals. PMID- 15117583 TI - Advances in the treatment of diabetic foot infections. AB - Lower extremity infections are frequent causes of substantial morbidity and mortality in the diabetic population, and these infections consume a large portion of resources expended on diabetic complications. Gram-positive cocci, particularly Staphylococcus aureus, are the most important pathogens in diabetic foot infections. These organisms are predominant both in mild infections (which are often monomicrobial), as well as in more severe and chronic infected wounds that more often have a polymicrobial cause. Appropriate clinical assessment and culturing of infections are critical in establishing the presence and severity of infection, in detecting osteomyelitis, and in directing the optimal treatment approach. Following necessary debridement and other surgical interventions (e.g., bone resection, revascularization), appropriate antibiotic therapy is a cornerstone of managing the infected lower extremity. Peripheral vascular (i.e., arterial) insufficiency and the increasing prevalence of antibiotic resistance are primary barriers to successfully managing these infections. Fortunately, alternative delivery systems (e.g., antibiotic beads, impregnated sponges) and novel antibiotics (e.g., levofloxacin, linezolid) are providing possible solutions to the challenges posed by this physically, emotionally, and financially devastating condition. PMID- 15117584 TI - Durability of glycemic control: a feature of the thiazolidinediones. AB - Type 2 diabetes is a rapidly growing disorder that affects millions of Americans. It usually results from a combination of insulin resistance and a beta-cell secretory defect leading to hyperglycemia and microvascular and macrovascular complications, including cardiovascular disease. With the increasing number of options available for the treatment of type 2 diabetes, it can be difficult to determine which medication to prescribe for each patient. Ideally, an agent that effectively lowers glucose concentrations while also minimizing disease progression should be chosen. The thiazolidinediones (TZDs), a relatively newer class of antidiabetic agents, have been shown to be efficacious in lowering glucose concentrations, maintaining glycemic control, and improving other cardiovascular risk factors. These include reduction of visceral adiposity, alteration of lipoprotein concentrations with a favorable distribution of cholesterol subfractions, and decreasing markers of inflammation and endothelial dysfunction. Overall, the TZDs appear to be a promising therapeutic option for consistent control of glucose levels and may slow the progression of type 2 diabetes. PMID- 15117585 TI - Fluorescence resonance energy transfer-based near-infrared fluorescence sensor for glucose monitoring. AB - A novel near-infrared (NIR) fluorescence affinity sensor for continuous glucose monitoring was developed and characterized. The sensor operates by fluorescence resonance energy transfer between a NIR chromophore linked to concanavalin A (ConA) and an NIR fluorophore linked to free dextran. The binding of dextran with ConA in the absence of glucose results in low fluorescence due to quenching; however, the quenching is reversed by competitive displacement of dextran from ConA by glucose. In order to increase thermodynamic stability and the lifetime of the sensor, ConA was immobilized within a macroporous bead matrix. The sensor was contained within a sealed hollow dialysis fiber (o.d. 215 microm, wall thickness 20 microm), preventing the macromolecules from leaking out and enabling glucose to rapidly enter the fiber lumen. A glucose-insensitive reference fluorophore was also incorporated to allow for ratiometric measurements, resulting in a robust sensor output that is independent of positional and/or light intensity changes. The response of the fluorescence affinity sensor to glucose was tested continuously in an automated test chamber at 37 degrees C. The sensor showed good dynamic range within physiologically relevant glucose concentration range (15% change over 2.5-30 mM, no hysteresis), fast response time (2-4 min), and a remarkable long-term stability (6 months). We interpret the improved longevity of this sensor to be the result of an optimized photo exposure regime and immobilization of ConA to the matrix. Its small size, ratiometric output, and NIR fluorescence make this sensor well suited for dermal implantation and continuous transdermal monitoring. PMID- 15117586 TI - Novel biosensors for long-term in vivo physiological monitoring. PMID- 15117591 TI - Response to Biermann: pilot randomised controlled trial results for evaluating the teaching utility of the AIDA diabetes software simulator--an overview. PMID- 15117592 TI - Daily glucose trend analyses and treatment adjustments using the continuous glucose monitoring system (CGMS, Medtronic MiniMed): "tweaking". PMID- 15117595 TI - Can serum HCG values be used in the differential diagnosis of pregnancy complicated by hypertension? AB - OBJECTIVE: The aim of our study is to determine whether the serum human chorionic gonadotropin (hCG) level is helpful in the differential diagnosis and in the clinical management and follow-up of preeclampsia, superimposed preeclampsia, and chronic hypertension during the third trimester. MATERIAL AND METHODS: Eighty hypertensive pregnant patients, who had been hospitalized, and 25 normotensive pregnant patients, who attended the outpatient perinatology clinic in Zeynep Kamil Women and Pediatric Diseases Education and Research Hospital between June 2001 and September 2001 were enrolled in the study. These patients were evaluated in five groups: mild preeclamptic, severe preeclamptic, superimposed preeclamptic, chronic hypertensive, and normotensive groups. The geometric means of hCG levels of these groups were compared with each other and cutoff levels for differential diagnosis were determined. RESULTS: The geometric mean of hCG levels was established as 17,361.31 mIU/mL in the mild preeclamptic group, 49,817.59 mIU/mL in the severe preeclamptic group, 41,101.09 mIU/mL in the superimposed preeclamptic group, 12,558.57 mIU/mL in the chronic hypertensive group, and 9647.98 mIU/mL in the normotensive group. When the geometric mean of the severe preeclamptic group was compared with the results of the normotensive patients, mild preeclamptic patients, chronic hypertensive patients, and superimposed preeclamptic patients, the mean hCG value of severe preeclamptic group was statistically significantly higher than all of the other groups (p < 0.001) except for the latter. The geometric mean of hCG levels of severe preeclamptic patients was compared with the geometric mean of hCG levels of superimposed preeclamptic patients (p > 0.05). The geometric mean of hCG levels in the chronic hypertensive group was lower than that of the superimposed preeclamptic group and the difference was statistically significant (p < 0.001). The geometric mean of hCG levels of the chronic hypertensive group was not significantly different from the results of the mild preeclamptic group and the normotensive group. There was, however, a statistically significant difference between the geometric means of hCG levels of mild preeclamptic patients and normotensive group (p < 0.001). The cutoff value of hCG was determined as 25,000 mIU/mL in differentiation of chronic hypertension from the severe preeclampsia, as 20,000 mIU/mL in differentiation of chronic hypertension from the superimposed preeclampsia, and as 30,000 mIU/mL in differentiation of severe preeclampsia from mild preeclampsia. CONCLUSION: The maternal serum hCG level is a useful laboratory tool when managing and treating hypertensive disorders that complicate pregnancy. The serum hCG level is especially significant in severe preeclampsia and superimposed preeclampsia. Therefore, a high serum hCG level can be a helpful marker in the diagnosis and clinical management by preventing possible complications resulting from severe and superimposed preeclampsia. PMID- 15117596 TI - An unusual cause of hypertension in pregnancy. AB - Aortic coarctation is an unusual cause of hypertension in pregnancy. We report the case of a 34-year-old woman with severe hypertension after surgical repair of aortic coarctation in childhood. An MRI showed a residual stenosis of the aortic arch and a small aneurysm. Pregnant postcoarctectomy patients are at an increased risk for developing hypertension during pregnancy due to residual aortic gradients and abnormal vascular reactivity of the precoarctation vessels. Women after repair of aortic coarctation should be closely monitored for blood pressure during pregnancy. PMID- 15117597 TI - Preeclampsia with abnormal liver function tests is associated with cholestasis in a subgroup of cases. AB - OBJECTIVE: To investigate whether women with preeclampsia and abnormal liver function tests have raised serum bile acids. DESIGN: Measurement of serum bile acids in serum specimens collected at the John Radcliffe Hospital, Oxford. SETTING: Imperial College School of Medicine. SAMPLE: Stored sera from 37 women with preeclampsia and abnormal liver function tests and from 19 controls. METHODS: Enzymic total bile acid assay. MAIN OUTCOME MEASURES: Total bile acid levels. RESULTS: Women with preeclampsia and abnormal liver function tests had higher median bile acid levels than controls (5.7 vs. 3.2, p = 0.01). The reason for the raised median serum bile acid levels in the patient group is that three (8%) women with preeclampsia had markedly raised serum bile acids levels. There were no obvious clinical or biochemical features specific to these patients. CONCLUSIONS: The pathological mechanisms causing hepatic impairment in some women with preeclampsia may predispose to cholestasis. As some women with preeclampsia and abnormal liver function complain of pruritus, we recommend checking the serum bile acids in this group of women. If these acids are raised the fetal prognosis may be adversely affected. PMID- 15117598 TI - Pregnancy outcome after previous pregnancy complicated by hepatic rupture. PMID- 15117599 TI - The effect of glyceryl trinitrate on hypertension in women with severe preeclampsia, HELLP syndrome, and eclampsia. AB - OBJECTIVE: The goal of this study is to evaluate the effect of glyceryl trinitrate (GTN) in the management of hypertension in women with preeclampsia, eclampsia, and HELLP syndrome. STUDY DESIGN: Fifty five women with preeclampsia, eclampsia, and HELLP syndrome administered GTN infusion for the management of hypertension were studied. Demographic, clinical, and perinatal outcome findings were collected for analyses. We recorded initial and maintenance doses of GTN, and duration of its use in prepartum and postpartum periods. We collected systolic and diastolic blood pressures (BPs) at admission and before the administration of GTN infusion. During the GTN infusion, we calculated average diastolic and systolic blood pressures 6 hours apart on the first day, 12 hours apart on the second day, and 24 hours apart on the third day. RESULTS: Of 55 women, 24 with severe preeclampsia, 16 with HELLP syndrome, and 15 with eclampsia were included in this study. In severe preeclampsia group, GTN infusion significantly reduced systolic and diastolic BPs beginning from the second quarter and third quarter, respectively, of first day (p < 0.05). In the HELLP syndrome group, GTN infusion significantly decreased systolic and diastolic blood pressures beginning from the third quarter and second quarter, respectively, of the first day (p < 0.05). In the eclampsia group, GTN infusion significantly reduced systolic and diastolic blood pressures beginning from the third quarter and first quarter, respectively, of the first day (p < 0.05). CONCLUSION: In women with severe preeclampsia, eclampsia, and HELLP syndrome, infusion of GTN can be used as an alternative agent to well-known drugs and causes no significant adverse effect to the mother and fetus. PMID- 15117600 TI - The effects of preeclampsia and oxygen environment on endothelial release of matrix metalloproteinase-2. AB - BACKGROUND: There is evidence of altered vascular endothelial function in women with preeclampsia as well as in the endothelial cells from umbilical vessels of preeclamptic pregnancies. Matrix metalloproteinase (MMP)-2 is elevated in the plasma of preeclamptic women and is a mediator of vascular reactivity; however, whether MMP-2 release is altered in preeclamptic endothelial cells is unknown. We hypothesize that MMP-2 release is enhanced in endothelial cells from preeclamptic compared with uncomplicated pregnancies and that this phenomenon may be mediated by an oxygen-dependent mechanism. Our specific hypothesis is that cells from normal pregnancies will demonstrate enhanced MMP-2 release at low oxygen (< 0.5%, 2%) compared to high oxygen (20%), thus mimicking the behavior of preeclamptic cells. METHODS: Human umbilical vein endothelial cells (HUVECs) from preeclamptic pregnancies (n = 4) and normal pregnancies (n = 4) were incubated for 12 hr in standard culture conditions (20% oxygen). In a separate series of experiments, HUVECs from normal pregnancies (n = 6) were incubated for 12 hr at < 0.5%, 2%, and 20% oxygen. Supernatants were analyzed for MMP-2 and tissue inhibitors of metalloproteinases (TIMP)-1 and -2. RESULTS: The HUVECs from women with preeclampsia demonstrated significantly enhanced release of MMP-2 (p < 0.05), TIMP-1 (p < 0.001), and TIMP-2 (p = 0.01) compared to normal cells. MMP-2 release from HUVECs from uncomplicated pregnancies was significantly elevated at 2% oxygen compared to < 0.5% and 20% oxygen (p < 0.05). TIMP-1 and -2 secretion was not altered with varying oxygen. CONCLUSIONS: Preeclamptic endothelial cells demonstrate significantly enhanced MMP-2, TIMP-1 and TIMP-2 release compared to normal cells. Our data show that there are significant effects of oxygen tension on MMP-2 release from normal cells; however, the magnitude of the enhanced release is small when compared to the differences in MMP-2 release in cells from preeclamptic and normal pregnancies. Furthermore, TIMP-1 and -2 release is not affected by changes in oxygen. It is unlikely that oxygen is a key mediator of the enhanced MMP-2, TIMP-1 and TIMP-2 release observed in preeclamptic cells. PMID- 15117601 TI - A survey of Canadian practitioners regarding the management of the hypertensive disorders of pregnancy. AB - BACKGROUND: How Canadian practitioners are managing the hypertensive disorders of pregnancy (HDP) is not known, particularly in relation to the 1997 guidelines published by the Canadian Hypertension Society (CHS). METHODS: A survey, with French and English versions (and covering diagnosis, evaluation, and management of pregnancy hypertension), was mailed to all members of the Society of Obstetricians and Gynaecologists of Canada (SOGC) (N = 1757, including obstetricians, family doctors practicing obstetrics, and midwives). Additionally, internists [i.e., all nephrologists (N = 191) and a random sample of 25% of general internists (N = 450)] registered with the Royal College of Physicians and Surgeons of Canada were sampled. The survey was distributed in two mailings and one reminder card. Data were entered into Microsoft Access, and Graph Pad Prism used to summarize responses [N (%)]. Differences in practice between specialties were examined, with a Bonferroni correction used to calculate a significant p value based on the number of comparisons and alpha of 0.05. RESULTS: Respondents numbered 1187 (49.5%), with 466 not informative for the purpose of the study (due to retirement, or practices that do not include pregnant women with hypertension). The final analysis included 721 completed surveys. For all types of HDP, most internists, family doctors, and midwives initiate nonpharmacological therapy (most common advice to quit work) at dBP 80-89 mmHg (i.e., primary prevention). Only for preeclampsia do obstetricians most frequently use this threshold; otherwise, dBP 90-99 mmHg is usually chosen. For nonsevere hypertension, antihypertensive drug therapy (most commonly methyldopa or labetalol) is started by most practitioners at dBP 90-99 mmHg, although obstetricians are more likely to choose a higher threshold (p < 0.0001). There is little agreement about dBP treatment goal; most internists and family doctors normalize dBP, whereas obstetricians appear to be divided on dBP goals of 80-89 (46-51%) vs. 90-99 mmHg (41-44%) for all HDP (p = 0.66). Severe hypertension is commonly treated with parenteral hydralazine, labetalol, or magnesium sulphate. Short-acting or sustained release nifedipine is used rarely/never by most practitioners. Approximately one-third of obstetricians and family doctors use diazepam to treat eclampsia. The vast majority use MgSO4 prophylactically in women with preeclampsia. INTERPRETATION: This survey has clarified current stated management of women with HDP, and identified the need for both research into the dBP treatment goal that optimizes pregnancy outcomes among women with HDP, and translation of definitive studies into clinical practice. PMID- 15117602 TI - Effects of maternally administered dexamethasone and acute hypoxemia at 0.7 gestation on blood pressure and placental perfusion in sheep. AB - Glucocorticoid administration to women in premature labor significantly decreases preterm infant morbidity and mortality. Fetal exposure to maternally administered glucocorticoids in late gestation causes fetal hypertension. We determined the effects of a single course (4 injections at 12-hr intervals) of dexamethasone (DM; 2 mg, a weight-adjusted dose equivalent to one-third the dose administered to pregnant women) or saline (S) in sheep at 103-104 days of gestation (dGA; term 149 dGA) on maternal and fetal blood pressure (BP). We also determined the BP and placental perfusion effects of acute maternal hypoxemia. Venous and arterial catheters were placed in 10 ewes and fetuses (DM = 6, S = 4) at 96 +/- 1 dGA. Maternal and fetal placental perfusion was determined with fluorescent microspheres. Dexamethasone increased fetal but not maternal BP; maternal and fetal placental blood flow and vascular resistance (VR) were unchanged. At 105 dGA, hypoxemia was induced for 1 hr by maternal nitrogen gas inhalation to decrease fetal PaO2 by 40%. Hypoxemia increased BP in DM but not S fetuses or mothers in either group. Hypoxemia decreased maternal placental blood flow by 39 +/- 7% and 51 +/- 9% and increased maternal placental VR by 65 +/- 7% and 69 +/- 6% in S and DM mothers, respectively. Hypoxemia did not alter fetal placental blood flow or VR in either treatment group. In summary, at 0.7 gestation, DM induces a hypertensive response to fetal hypoxemia that is characteristic of older fetuses but does not alter hypoxemia-induced reductions in maternal placental blood flow. PMID- 15117603 TI - Evidence of increased oxidative stress, unexplained by lipid changes, is present in nulliparous black women from early gestation. AB - OBJECTIVE: We tested the hypothesis that race-specific lipid changes in pregnancy could predispose to oxidative stress and might increase the risk of preeclampsia for black women. STUDY DESIGN: Blood samples were obtained at five time points in pregnancy and postpartum for 15 black and 15 white women with normal pregnancy. Serum or plasma samples were analyzed for cholesterol, triglycerides, LDL cholesterol, HDL total and subfractions, free fatty acids, uric acid, and malondialdehyde (MDA). RESULTS: Cholesterol and LDL-cholesterol values were higher in white women than in black women (RMANOVA, p = 0.04) while MDA values were higher in blacks throughout pregnancy (RMANOVA, p = 0.03). Although not significantly different, black women had lower mean triglycerides, while mean total HDL and subfractions HDL2 and HDL3 were higher for black women. Both races evidenced smaller, denser LDL particle size with advancing gestation and a return to early pregnancy LDL particle size by 6 weeks' gestation. CONCLUSION: These data do not support quantitative lipid change as the explanation for the increased incidence of preeclampsia in black women. Despite this, black women manifest increased evidence of oxidative stress, postulated to be an important contributor to the pathogenesis of preeclampsia from early gestation. PMID- 15117604 TI - Prospective analysis of placenta growth factor (PlGF) concentrations in the plasma of women with normal pregnancy and pregnancies complicated by preeclampsia. AB - OBJECTIVE: The aim of this study was to analyze if levels of plasma PlGF in the second half of pregnancy have predictive value for the identification of women destined to develop preeclampsia or another complication of pregnancy. MATERIAL AND METHODS: A bank of 1.543 randomly collected plasma samples (22-29 weeks of gestation) was established and PlGF concentrations were quantitated in a prospective longitudinal study in all pregnant women who developed a complication of pregnancy in late gestation (177 of 1.543) and the same number of gestational age matched pregnancies with normal outcome. RESULTS: Plasma PlGF levels in pregnant women rise steadily throughout pregnancy from the level of nonpregnant women (< 50 pg/mL) to levels exceeding 500 pg/mL after 30 weeks of gestation. Just 7.3% of pregnant women with normal outcome of pregnancy had PlGF levels of less than 200 pg/mL beyond 22 weeks of gestation (3.7% beyond 25 weeks of gestation). The rise in plasma PlGF in the second half of pregnancy was significantly attenuated in pregnancies that were complicated by preeclampsia in late gestation. Of all women who developed preeclampsia, 27.3% (12 of 44) had plasma PlGF levels below 200 pg/mL. The attenuation of the rise in plasma PlGF was not evident in other complications of pregnancy (transient hypertension, fetal retardation, pregnancy diabetes, premature contractions, proteinuria without hypertension, infections during pregnancy). CONCLUSION: The rise in plasma PlGF levels observed in normal pregnancies is significantly attenuated in pregnancies complicated by preeclampsia. Yet, due to the low sensitivity and specificity, plasma PlGF levels in the second half of pregnancy have no predictive value for the identification of individual women destined to develop preeclampsia. PMID- 15117605 TI - Analysis of the S810L point mutation of the mineralocorticoid receptor in patients with pregnancy-induced hypertension. AB - OBJECTIVE: A missense mutation at codon 810 (Ser --> Leu) of the mineralocorticoid receptor was recently observed in a family with early manifestation of hypertension. Our objective was to determine if this mineralocorticoid receptor alterations is prevalent in patients with pregnancy induced hypertension. METHODS: Thirty-eight women with hypertension during pregnancy were tested for the mineralocorticoid receptor gene mutation. DNA was extracted out of blood leucocytes. PCR and automated DNA sequencing were used to analyze exon 6 for the S810L missense mutation. Anamnestical data concerning cardiovascular risk factors and family history were evaluated with a questionnaire. Pregnancy course and outcome were documented in all cases. RESULTS: In 33 patients with pregnancy-induced hypertension and in five patients with exacerbation of preexisting hypertension in pregnancy no point mutations were found at codon 810 in exon 6. CONCLUSIONS: Our data suggest that the S810L missense mutation of the mineralocorticoid receptor does not play a major role in the etiology of pregnancy-induced hypertension in a German /Turkish population. PMID- 15117606 TI - Septic pelvic thrombophlebitis and preeclampsia are related disorders. AB - OBJECTIVE: To elicit factors associated with the postpartum development of septic pelvic thrombophlebitis in a single large referral tertiary patient population. METHODS: A nine-year single institution retrospective case review of all patients with enigmatic fever and septic pelvic thrombophlebitis was analyzed. RESULTS: A total of 55 patients with septic pelvic thrombophlebitis were provided care during the study interval. The average gestational age at delivery was 36.8 +/- 4.3 weeks. The most prevalent concurrent medical complication of pregnancy was preeclampsia (45%) while chorioamnionitis affected only 13%. The average length of ruptured membranes was 22.8 +/- 56.8 hours (median 10.5, 95% confidence interval [CI] 7.0-38.7 hours), with 22% of patients undergoing amnion rupture at the time of cesarean delivery. Prolonged (>24 hours) amnion rupture occurred in only 9% of patients. Most affected patients were delivered abdominally (91%) but a minority delivered vaginally (9%). Antibiotic therapy for presumed infection was initiated at 27.4 +/- 24.6 hours postpartum. Subsequently intravenous heparin therapy was initiated 128.9 +/- 54.2 hours thereafter enigmatic fever defervesed 37.2 +/- 36.8 hours later (median 34.0, 95% CI 27.2-47.3 hours). Patients received 6.3 +/- 1.8 days of heparin therapy. CONCLUSION: In this series, septic pelvic thrombophlebitis was frequently preceded by cesarean delivery and commonly associated with preeclampsia. Unexpectedly, a small number of patients suffered prolonged rupture of membranes or chorioamnionitis. We speculate that the cesarean delivery of a population of at-risk patients with preeclampsia may predispose them to develop septic pelvic thrombophlebitis. PMID- 15117607 TI - [Proteinuria and urinary beta 2-microglobulin as markers of tubular malfunction in the assessment of severity of acute pancreatitis]. AB - INTRODUCTION: Proteinuria is a common finding in acute pancreatitis (AP). Increased urinary beta 2-microglobulin can be explained by renal tubular malfunction induced by substances released from the pancreas. The degree of renal tubular malfunction may reflect the severity of AP. AIM: To assess proteinuria and urinary beta 2-microglobulin as prognostic factors in AP. PATIENTS AND METHODS: We retrospectively studied patients with AP with symptom onset within 24 hours before admission. Random urine specimens were obtained on days 1, 2 and 3 after admission. In a subgroup of 25 patients, urine samples could be obtained within 24 hours of symptom onset on day 1. The severity of AP was established using the Atlanta criteria. Proteinuria and beta 2-microglobulin were determined and were adjusted by urinary creatinine concentrations. RESULTS: We studied 51 patients with AP (26 men and 25 women; age: 59.6 (+/-16.7 years). Fifteen cases of AP were severe and 36 were mild. The most frequent etiology was gallstones (60.1%). Levels of proteinuria were (median and interquartile range) in mg/g creatinine: day 1: 180.5 (84.0-250.9), day 2: 164.3 (16.7-421.7), and day 3: 136.7 (24.0-371.29). Differences between severe and mild AP were significant on day 2 of admission: 339.7 (191.7-471.8) versus 120,1 (11.0-382.6); p = 0.04. Levels of urinary beta 2-microglobulin in AP on days 1 to 3 postadmission were: 9.7 (1.1-93.3), 27.6 (4.7-421.4) and 88.3 (7.3-415.2) microg/mg of creatinine, respectively. When urinary beta 2-microglobulin was compared between severe and mild AP, no significant differences were found among days 1, 2 and 3. Selection of only the subgroup of patients whose urine samples were obtained within 24 h of symptom onset, did not improve the results of these urine markers for the group as a whole. CONCLUSIONS: 1) Proteinuria was slightly increased in severe AP and was able to discriminate between mild and severe episodes on day 2 of admission. 2) Urinary beta 2-microglobulin as a tubular malfunction marker did not discriminate between mild and severe AP in patients in our study. PMID- 15117608 TI - [Burden of gastroesophageal reflux disease. Evidence from a recent methodological study in Spain]. AB - INTRODUCTION: The clinical and socioeconomic burden of gastroesophageal reflux disease (GERD) is considerable. AIM: The aim of this study was to describe the impact of heartburn on patients' health-related quality of life (HRQL) in Spain, using validated generic and disease-specific instruments to measure patient reported outcomes. PATIENTS AND METHODS: Patients with symptoms of heartburn completed the Spanish versions of the Gastrointestinal Symptom Rating Scale (GSRS), the Quality of Life in Reflux and Dyspepsia questionnaire (QOLRAD), the Short Form-36 (SF-36) and the Hospital Anxiety and Depression (HAD) scale. The frequency and severity of heartburn in the previous 7 days were also recorded. RESULTS: One hundred fifty-eight patients completed the assessments (mean age: 51 years, SD = 16; 58% female). Sixty-one percent of the patients had moderate symptoms and 73% had symptoms on three or more days in the previous week. Patients were most bothered by symptoms of reflux (mean GSRS score of 3.8 on a scale of 1 [not bothered] to 7 [very bothered]), abdominal pain (3.1) and indigestion (3.1). As a result of their symptoms, patients experienced problems with food and drink (mean QOLRAD score of 4.5 on a scale of 1 to 7, where 1 represents the most severe impact on daily functioning), impaired vitality (4.7) and emotional distress (4.8). This led to impaired overall HRQL across all domains (mean SF-36 score of this heartburn population compared with a general population in Spain). The HAD scale showed that 28% of patients were anxious and 13% were depressed. CONCLUSION: There is consistent evidence that GERD substantially impairs all aspects of HRQL. PMID- 15117609 TI - [Wilson's disease with severe neurological manifestations: response to trientine plus zinc therapy]. AB - In patients with Wilson's disease and neurological manifestations, treatment with D-penicillamine can cause worsening of neurological symptoms, usually in the first few weeks of treatment. Because the neurological damage can be severe and irreversible, the use of D-penicillamine is controversial, and several authors believe that it should be avoided. Studies of the use of ammonium tetrathiomolybdate as an alternative chelating agent for the initial treatment of neurologic Wilson's disease are still in the experimental phase. Published experience on the simultaneous use of trientine, another chelating agent, and zinc, which blocks intestinal absorption of copper, is promising but limited. We present the case of a 17 year-old boy with severe neurologic Wilson's disease that had first presented six years previously. The patient showed a complete recovery after six months of treatment with a combination of trientine and zinc acetate. PMID- 15117610 TI - [Multiple angiodysplasia of the small intestine. A diagnostic and therapeutic challenge]. AB - Small bowel bleeding is infrequent and presents a challenge to the clinician. Approximately 30-40% of gastrointestinal bleeding localized in the small bowel is due to angiodysplasia, a vascular malformation. We present the case of a patient with multiple angiodysplasia of the small bowel who required push enteroscopy and capsule endoscopy to establish the diagnosis. Treatment with subcutaneous octreotide was successful. In conclusion, in doubtful cases or in patients with persistent hemorrhage, capsule endoscopy can improve the diagnostic yield of enteroscopy in bleeding gastrointestinal vascular lesions such as angiodysplasia. Endoscopic treatment (laser coagulation) and drug therapy (somatostatin or analogs) are valid alternatives in inoperable or non-resectable cases. PMID- 15117611 TI - [Familial hepatic hemangiomas]. AB - Cavernous hemangioma is the most frequent benign hepatic tumor. These tumors are usually small, solitary or multiple, and asymptomatic but they can be large. In the majority of cases, these tumors are sporadic. We describe a family in which six members from three successive generations presented cavernous hepatic hemangiomas. One of the women, who was taking oral contraceptives, presented a giant cavernous hemangioma causing pain in the right hypochondriac region and biochemical abnormalities while the remaining cases were asymptomatic. We found only one study in the literature that reports familial cavernous hepatic hemangioma; a further two studies describe hepatic hemangiomas in members of families with angiomas in other sites. We believe that description of a new family with this disease is of interest. The findings could support autosomal dominant inheritance as the mode of transmission of these tumors, as well as the existence of proliferative factors in these tumors. PMID- 15117612 TI - [Liver abscess and Crohn' disease. Report of 3 cases]. AB - Liver abscess is a rare complication of Crohn's disease. Its prevalence and mortality are higher in patients with Crohn's disease than in the general population. Owing to its nonspecific clinical presentation, which may be mistaken for reactivation of Crohn's disease or be masked by simultaneous steroid therapy, a high index of suspicion is required for an early diagnosis and prompt treatment. We report 3 cases of Crohn's disease complicated with liver abscess in which the only common features were the absence of clinical or even endoscopic activity of Crohn's disease at diagnosis and the presence of an anastomotic leak due to right ileocolectomy in the previous year. In all patients, outcome was satisfactory with antibiotic therapy and percutaneous catheter drainage. PMID- 15117613 TI - [Molecular mechanisms in bile formation]. PMID- 15117614 TI - [Latin-American consensus document on irritable bowel syndrome]. PMID- 15117615 TI - [Methotrexate in the treatment of autoimmune hepatitis]. PMID- 15117617 TI - [Inferior vena cava filter update]. PMID- 15117616 TI - [Massive lower gastrointestinal bleeding treated with superselective embolization]. PMID- 15117618 TI - [Prevalence of sleep-disordered breathing in patients with acute ischemic stroke: influence of onset time of stroke]. AB - OBJECTIVE: To analyze the prevalence of sleep-disordered breathing in patients with acute ischemic stroke and the influence of the characteristics of the stroke and time of onset. PATIENTS AND METHODS: Polysomnography was performed with an Autoset Portable Plus II in 139 patients within 72 hours of the onset of symptoms. Standard polysomnographic data, signs and symptoms related with sleep apnea-hypopnea syndrome (SAHS) prior to ischemic stroke, vascular risk factors, and characteristics and onset time (day/night) of ischemic stroke were recorded. The polysomnographic data were compared with results published for subjects of a similar age in the general population. RESULTS: The mean age was 73.6 (SD 11.1) years (59% of the patients were men). Prior to the stroke, 64.7% of the patients snored, 21.6% presented repetitive sleep apneas, and 35.6% had daytime sleepiness. The mean apnea-hypopnea index (AHI) was 29.1 (17.9) episodes/hour, the obstructive component of which was 20.1 (15.7) episodes/hour. Five patients presented Cheyne-Stokes breathing. The AHI (for all cut-points from 5 to 50), chronic snoring, and daytime sleepiness were significantly greater than those published for the general population. The stroke characteristics showed no significant differences between daytime and nighttime onset. Nighttime stroke (60.4%) was associated with a significantly higher AHI (33.3 compared to 24.7 episodes/ hour) mainly because of obstructive apneas. Nighttime stroke was also associated with a greater nighttime desaturation and a greater probability of SAHS symptoms prior to stroke (odds ratio, 2.62). In contrast, there were no differences in vascular risk factors between daytime and nighttime stroke onset. CONCLUSION: The prevalences of sleep-disordered breath- ing with clinical signs and symptoms of SAHS were high in this population of patients with acute ischemic stroke. Patients with nighttime stroke had more obstructive sleep-disordered breathing and a higher clinical probability of obstructive SAHS before stroke. These findings support the hypothesis that obstructive SAHS is a risk factor for ischemic stroke, particularly for strokes presenting at night. PMID- 15117619 TI - [Pleural mesothelioma: experience with 62 cases in 9 years]. AB - OBJECTIVES: To describe the diagnostic approach, clinical and radiological characteristics, and survival of patients with pleural mesothelioma treated in our hospital over a 9-year period. PATIENTS AND METHOD: All patients with a diagnosis of pleural mesothelioma diagnosed in our hospital from January 1992 through December 2000 were studied. RESULTS: Sixty-two patients (49 men) with a mean age of 65 years (range, 45-85) were diagnosed. Probable or known contact with asbestos was established for 41 patients (66%). Ninety-four percent of the patients had chest pain or dyspnea at the onset of clinical assessment. The tumor was situated in the right hemithorax in 33 patients; 59 patients had pleural effusion, and 3 only had pleural thickening. The pleural fluid was bloody in 19% of patients, glucose levels were less than 60 mg/dL in 44%, and the pH of pleural fluid was less than 7.20 in 19%. The diagnosis was established by pleural biopsy for 52%, and by thoracoscopy or thoracotomy for 44%. The median survival was 11 months (95% confidence interval, 8-15); the probability of survival was 0.22 after 2 years, and 0.09 after 5. For the subgroup of patients with epithelial tumors the probability of survival was 0.31 after 2 years and 0.16 after 5 years. In the univariate analysis the predictors of survival were general clinical status (Karnofsky scale), platelet count, serum albumin level, pleural pH, glucose and lactate dehydrogenase levels, and histological type. CONCLUSIONS: The clinical, radiological, and biochemical characteristics of the pleural fluid from patients with pleural mesothelioma and their survival rate were described. PMID- 15117620 TI - [Analysis of respiratory muscle structure and tumor necrosis and insulin-like growth factor expression in chronic obstructive pulmonary disease: are samples valid if obtained during thoracotomy performed because of localized pulmonary neoplasia?]. AB - OBJECTIVE: Various methods have been used to obtain samples to study the structure of human respiratory muscles and the expression of diverse substances in them. Samples are most often obtained from autopsies, from muscle biopsies during thoracotomy performed because of a localized pulmonary lesion (TLL), and from ambulatory thoracoscopic biopsy in patients free of comorbidity (AT). The disadvantage of the first 2 of these methods lies in the possibility of interference from factors related to the patient's death in the first case or from the disease that necessitated surgery in the second. Although AT is free from the disadvantages of the other 2 methods, it is impossible to obtain samples of the diaphragm the principal respiratory muscle with this procedure. The objective of this study was to analyze the fibrous structure of the external intercostal muscle of patients with chronic obstructive pulmonary disease and to quantify the expression of the principal inflammatory cytokine tumor necrosis factor alpha (TNF-alpha)- and of insulin-like growth factor (IGF-1) in the same muscle, comparing the results obtained with TLL and AT samples. METHODS: Prospective and consecutive samples were taken of the external intercostal muscle (fifth space, anterior axillary line) in 15 patients with chronic obstructive pulmonary disease (mean [SD] age 66 [6] years; forced expiratory volume in 1 second 49% [9%] of predicted; PaO2 75 [9] mm Hg). Samples were taken during TLL (8 patients, all with pulmonary neoplasms but carefully selected in order to rule out systemic effects) or TA (7 patients). Patients with serious comorbidity were excluded from the second group. Samples were processed for structural analysis of fibers (immunohistochemical and enzymatic histochemical) and genetic expression of TNF-alpha and IGF-1 (real-time polymerase chain reaction). RESULTS: No differences in the structure of fibers were found between the 2 groups. No differences were observed in the expression of TNF-alpha or IGF-1. CONCLUSIONS: Using rigorous criteria, the TLL method appears to be suitable for studying the structural characteristics and expression of inflammatory cytokines and growth factors in the external intercostal muscle. Moreover, it can also be inferred that TLL is probably also useful for obtaining samples of the diaphragm, a muscle which cannot currently be sampled by any alternative method. PMID- 15117621 TI - [Bronchial carcinoid tumor: a retrospective analysis of 62 surgically treated cases]. AB - OBJECTIVE: To evaluate the results of surgical treatment for lung carcinoid tumor. PATIENTS AND METHOD: The medical records of 62 patients who underwent surgical intervention for lung carcinoid tumor between May 1985 and October 2000 were reviewed. RESULTS: Fifty-two patients had typical carcinoid tumors and 10 had atypical carcinoid tumors. Hilar or mediastinal lymph node metastases were present in 9 patients. Distant metastasis occurred in 5 patients and was significantly more frequent in those with the atypical carcinoid histological subtype. The overall survival rate at 15 years was 70%, with a mean survival rate of 138 (SD 11) months, calculated with the Kaplan-Meier method. We found no statistically significant correlation between smoking and the development of carcinoid tumors. CONCLUSIONS: Although carcinoid tumors behave like low-grade malignant tumors, they should be treated in the same way as other malignant lung tumors. Curative surgical resection is the technique of choice whenever possible. PMID- 15117622 TI - [Nasal nitric oxide]. PMID- 15117623 TI - [Infection by influenza virus in childhood: a call for broader influenza vaccination]. PMID- 15117624 TI - [Video-assisted thoracoscopic surgery in 3 cases of adult cystic adenomatoid malformation]. AB - Congenital cystic adenomatoid malformation involving the lung is a rare hamartomatous condition that is usually diagnosed in the neonatal period. The presentation of this malformation in older patients is exceptional and usually manifests in a series of recurrent lung infections affecting a single lobe or segment. The treatment of choice is complete surgical exeresis. This report of 3 cases of late presentation focuses on the surgical approach used and the unusual manifestation of recurrent spontaneous pneumothoraces in 1 patient. The patients were females aged 15, 16, and 25 years with histories of various respiratory diseases (extrinsic asthma, recurrent pneumonias, and pneumothoraces). The patients were referred to us for surgery with suspected diagnoses that were different from the final diagnoses in all cases. All underwent diagnostic video assisted thoracoscopy to explore the affected hemothorax, and definitive treatment was possible during the procedure for 2 patients (a lobectomy and an atypical segmentectomy) by video-assisted surgery. The third patient underwent lobectomy by lateral thoracotomy after exploratory video-assisted thoracoscopy. Short- and long-term outcomes were excellent for all 3 patients. PMID- 15117625 TI - [Bronchiolitis obliterans associated with paraneoplastic pemphigus: a paraneoplastic autoimmune multiorgan syndrome]. AB - INTRODUCTION: Paraneoplastic pemphigus is a mucocutaneous disease characterized by well defined clinical and immunopathological features associated with neoplasia. Recent evidence of bronchial epithelium involvement has led to the suggestion that this process is a paraneoplastic autoimmune multiorgan syndrome. CLINICAL OBSERVATION: We report the case of a patient with lichenoid eruptions on the skin and mucous membranes who later developed progressive dyspnea. With a suspected diagnosis of paraneoplastic autoimmune multiorgan syndrome, the following diagnostic tests were performed: histology and immunofluorescence of the skin, oral mucosa, and bronchial epithelium; indirect immunofluorescence of serum; pulmonary function tests; and evaluation for an occult neoplasm. Findings of pathology and immunofluorescence confirmed the suspected diagnosis. The computed thoracoabdominal tomography revealed signs of bronchiolitis and the presence of a retroperitoneal tumor. CONCLUSIONS: Awareness of the mucocutaneous manifestations of paraneoplastic autoimmune multiorgan syndrome, and confirmation of this diagnosis by simple laboratory techniques can facilitate the early detection of occult neoplasia and forestall respiratory involvement. PMID- 15117626 TI - [Facial ecchymosis in a thrombocytopenic patient after fiberoptic bronchoscopy]. PMID- 15117627 TI - [Pseudo-pancoast syndrome caused by a solitary fibrous tumor of the pleura]. PMID- 15117628 TI - [The most rapid and complete guide to primary care diagnosis]. PMID- 15117629 TI - [Information given to patients and their participation in clinical decision making]. AB - OBJECTIVES: To evaluate the information provided by physicians during consultations, the information requested by patients, and patients' participation in decision-making. DESIGN: Descriptive, cross-sectional study based on observations of visits to the doctor's office and subsequent telephone interview. SETTING: Primary care centers in Toledo, Spain.Participants. 152 clinical interviews.Interventions. Questionnaire with items on age, sex, types of information provided by the physician, information requested by the patient, and evaluation of the patient's participation in decision-making and degree of satisfaction. MAIN MEASURES AND RESULTS: Mean age of the patients was 41.1 years, and 55.9% were women. The type of information given most frequently to patients was related to treatment (88.3%). Of the 152 patients whose visits were observed, 55 (36.2%) did not request additional information. The information requested most frequently when not provided spontaneously by the physician concerned treatment (35.3%) and cause of the symptoms (29.7%). Almost all patients (94.0%) considered the information received to be sufficient. However, 22.7% stated that when they left the doctor's office there was something they wished they had asked about, and 18.6% said they understood the doctor's explanations "in part." According to the observers, 69.4% of the patients did not take part in the decision about their treatment. CONCLUSIONS: The information provided by the physician was not as complete as it might have been. Patients usually ask few questions, and a large percentage of patients had something they wished they had asked about, or did not fully understand the information. Patients' participation in decision making was low. PMID- 15117630 TI - [Commentary: The challenge of patient information]. PMID- 15117631 TI - [Cancer diagnosis: do we want to know the truth?]. AB - OBJECTIVE: To find people's desire for information and their other attitudes when faced with a cancer diagnosis. DESIGN: Transversal, descriptive study. SETTING: Irun, Onati and Asteasu Health Centres, Gipuzkoa, Spain. PARTICIPANTS: From the appointments for 19 clinics in March-May 2002, a randomised, prospective and systematic sample, stratified by lists, was extracted (n=725). A modified Fernandez Diaz questionnaire was administered. It had personal details and 8 questions with closed replies on aspects of communication (5) and attitudes to death (3). RESULTS: 81.3% (95% CI, 78.4-84.1) wanted to know their diagnosis. 68.9% (95% CI, 65.5-72.2) chose the doctor to tell them. Of these, 36.6% chose the specialist and 26.6%, the general practitioner. If the person affected was a family member, 46.6% (95% CI, 42.9-50.2) would always tell the person; 38.6% (95% CI, 35-42.1), only if the person asked; and 9.6%, never (95% CI, 7.4-11.7). 49.3% (95% CI, 45.6-52.9) would prefer to die at home. The greatest fear for 44% (95% CI, 40.3-47.6) was pain. CONCLUSIONS: Most people want to be informed of their diagnosis. Neither an urban or rural environment nor having had family members with cancer affects their view. If the person affected by cancer is a family member, very few people favour not telling him/her at all. Sufferers want the doctor to tell them the information, prefer to die at home and pain is what worries them most. PMID- 15117632 TI - [Sexual behaviour and use of condoms by adolescents in our environment]. AB - OBJECTIVE: To identify sexual behaviour in adolescents. DESIGN: Transversal, descriptive study, by means of an anonymous structured questionnaire. SETTING: Urban secondary school. PARTICIPANTS: Simple random sample of 84 students (4th year of ESO--c. 15/16 years old). MAIN MEASUREMENTS: Personal details, sexual practices, use of condoms. RESULTS: 61.9% were girls. Average age was 16.4 (95% CI, 16.2-16.6). They had a girl/boy-friend or casual partner (33.7%) and stable partner (60.7%). 82.2% had total or sufficient trust. Boys' first masturbation was between 14 and 16 (46.8%). 100% had done so by the age of 16. 72.3% of girls had never masturbated at this age (P<.001).Boys' first kiss on the lips occurred between 14 and 16 (46.8%); 100% had done so by 16. 60.7% of girls had done so between 14 and 16, and 92.2% by 16.30.1% of boys had had their first experience of coitus by the age of the questionnaire, whereas 22.5% of girls had. Both sexes used a condom on 71.4% of occasions.38.1% had sex with penetration, 46.2% masturbated. 22.2% had had 2 sexual partners, 8.9% had had 3, and 40.0% had had 4 or more. Condoms were used in 83.3% of relationships with penetration. CONCLUSIONS: This is a sexually active population, susceptible to sexually transmitted diseases (STD) and unwanted pregnancies, as it does not use condoms in every relationship involving penetration. Health education activities need to be encouraged. PMID- 15117633 TI - [Do we forget asthma as a chronic illness in our primary care consultations?]. AB - OBJECTIVE: To describe the characteristics of the asthmatic population at our centre through the information gathered from clinical records, the morbidity caused by asthma and the health education given. DESIGN: Retrospective, descriptive study. SETTING: Urban primary care team. PARTICIPANTS: Asthma sufferers over 14 registered at the centre (n=284). MAIN MEASUREMENTS: Review of the clinical records and analysis of data on diagnosis, follow-up and control of the illness. RESULTS: Prevalence of asthmatics at our centre ran at 1.8%. 73.6% (68.5-78.7) were women, whose average age was 51.25 (21.57). 19% were illiterate. The most common kind of asthma was intermittent (14.1%, 10-18.1). Respiratory infections sparked off acute attacks in 15% (11.3-19.7) of cases. 39.8% (31.4 45.5) (n=113) were diagnosed at the health centre. Spirometry was used as a diagnostic test in 23% (15.2-30.8) (n=26) of these 113 patients. The PC doctor participated in monitoring 81.7% (77.2-86.2) of asthma cases. Spirometry was used in the previous 3 years on 45.4% (39.6-51.2). 95% of spirometry tests were conducted at the hospital. As to health education, there was no record of explanations about the concept of asthma or of breathing techniques in 90.1% (86.1-93.4) and 81% (76.4-85.5) of clinical records, respectively. CONCLUSIONS: High percentage of asthmatics monitored by PC doctors. Under-recording in clinical records of asthma-related action taken. Few additional monitoring tests were conducted. There were few data on health education. PMID- 15117634 TI - [Regulation of residents working day as a prerequisite for quality training]. PMID- 15117635 TI - [Navigating on the internet in search of health information: all that glitters is not gold]. PMID- 15117637 TI - [Primary hypothyroidism in the elderly]. PMID- 15117638 TI - [Diagnosis of acute lymphoblastic leukaemia from chest pain]. PMID- 15117639 TI - [Cervical-arthritic myelopathy of sub-acute evolution]. PMID- 15117640 TI - [Keratosis plantar sulcatum]. PMID- 15117642 TI - [Tables of cardiovascular risk and evidence. In defence of the calibrated Framingham table]. PMID- 15117643 TI - [Global muscle dysfunction and exacerbation of COPD: a cohort study]. AB - BACKGROUND AND OBJECTIVE: The purpose of this study was to evaluate on a prospective fashion the effects of clinical relapses of chronic obstructive pulmonary disease (COPD) on both peripheral and respiratory skeletal muscle functions. PATIENTS AND METHOD: We included 49 patients (males, 63 [11] years) who were assigned to three cohorts: a) COPD patients who were hospitalized in a conventional ward because of a relapse of their disease; b) patients hospitalized in conventional wards because of another lung disease or a pulmonary nodule; and c) COPD patients whose disease was stabilized (outpatients). Sequential measurements were made by means of anthropometry, serum biochemistry and body bioimpedance (BIA). In COPD patients with a disease relapse, we assessed changes in the function of peripheral muscles [force (Fhand) and resistance (Tlimhand) of hands], inspiratory muscles (PImax) and respiratory muscles (PEmax). RESULTS: Patients were evaluated during a 6 [2] days period. Patients with a COPD relapse displayed a global and progressive functional muscle impairment, which was expressed as a decrease of PEmax (17 [12]%), F hand-D (6 [9]%), F hand-ND (7 [8]%), Tlim hand-D (28 [26]%) and Tlim hand-ND (23 [16]%). These changes showed a linear trend. BIA exhibited a loss of lean mass (7 [6]%, p < 0.05) which would have been unnoticeable if only the body weight was quantified. Pneumonia cases showed similar changes in BIA. On the other hand, the cohort of patients with stable COPD did not have changes in both muscle function and BIA. CONCLUSIONS: COPD exacerbation is associated with an acute and global impairment of the function of respiratory and peripheral skeletal muscles. It is possible that these changes are related to an acute loss of muscular mass (proteolysis). This muscle dysfunction is not detected if only the inspiratory muscular function is evaluated--possibly because of the coexistence of transitory mechanic factors. PMID- 15117644 TI - [Clinical benefit following the implementation of a specialized urgent stroke care system]. AB - BACKGROUND AND OBJECTIVE: Several strategies as stroke teams, stroke code teams, or stroke units development have demonstrated to improve stroke care quality. The potential benefit of implementing them as a whole has not been studied. We aimed to test the clinical efficacy of a specialized and urgent stroke assistance system in a University Hospital, as well as the specific impact of each part of the system on several clinical indicators. PATIENTS AND METHOD: The implementation of the system followed three consecutive steps: stroke team and stroke code development, stroke unit creation and finally on-call stroke neurologists incorporation. Several clinical indicators to evaluate results have been selected. We compared data available before system onset (1992-1997) with data obtained during the system implementation (1998-2002). Modification in the results indicators following each of the individual steps of the system was also evaluated. RESULTS: During the five years of the system implementation, 5843 stroke patients have been prospectively studied. Admission and readmission necessities were reduced up to 34.2% and 81.8% respectively. Length of stay progressively decreased from 18 (pre-1998) to 7 days (2002). In-hospital mortality and institutionalization necessities were reduced to 50.1% and 50.5% respectively. The third step, in which on-call stroke neurologist were incorporated to the system, has demonstrated to be the most efficient in decreasing the length of stay, hospital mortality and institutionalization necessities. CONCLUSIONS: The creation of a specialized urgent stroke care system, protocol based and developed in stroke units, improves the medical assistance quality for stroke patients. Stroke neurologists on-call have a relevant role in the system working. PMID- 15117645 TI - [Clinical significance of increased serum ferritin levels]. AB - BACKGROUND AND OBJECTIVE: We aimed to determine the diagnostic alternatives indicated by serum ferritin levels (2000 ng/ml, and to establish the clinical processes associated with very high levels (5000-10 000 ng/ml). PATIENTS AND METHOD: We retrospectively analyzed cases with serum levels of ferritin serum equal to or greater than 2000 ng/ml between March 2000 and November 2001. Data were obtained from the laboratory's computerized database. Patients' medical records were reviewed by means of a protocol which established the clinical conditions associated with these serum ferritin values. RESULTS: The study involved 135 patients with ferritin levels equal to or greater than 2000 ng/ml. Clinical syndromes included hematological diseases (45.9%), liver diseases (23%), chronic renal failure (17.78%), neoplastic diseases (10.4%), systemic inflammatory diseases (7.4%), chronic transfusions (7.4%), and non-HIV systemic infections (5.9%). Syndromes which are not usually associated with extreme serum ferritin levels were identified in 3.7% of the patients. The highest concentrations were seen in the systemic inflammatory disease group: 5856 (2492) ng/ml. Within this group, four patients with adult onset Still's disease (AOSD) displayed the highest mean ferritin levels: 11 322 (5474) ng/ml. CONCLUSIONS: Elevated serum ferritin levels act as a non-specific marker for a large number of disorders. In certain inflammatory diseases such as adult onset Still's disease (AOSD), this finding may be an important tool. PMID- 15117646 TI - [COPD: not just the airway matters]. PMID- 15117647 TI - [Stroke units: better patients attention, higher efficacy and lower cost]. PMID- 15117648 TI - [Dying peacefully: evaluation of its influencing factors]. PMID- 15117649 TI - [Obesity studies in candidate genes]. AB - There are more than 430 chromosomic regions with gene variants involved in body weight regulation and obesity development. Polymorphisms in genes related to energy expenditure--uncoupling proteins (UCPs), related to adipogenesis and insulin resistance--hormone-sensitive lipase (HLS), peroxisome proliferator activated receptor gamma (PPAR gamma), beta adrenergic receptors (ADRB2,3), and alfa tumor necrosis factor (TNF-alpha), and related to food intake--ghrelin (GHRL)--appear to be associated with obesity phenotypes. Obesity risk depends on two factors: a) genetic variants in candidate genes, and b) biographical exposure to environmental risk factors. It is necessary to perform new studies, with appropriate control groups and designs, in order to reach relevant conclusions with regard to gene/environmental (diet, lifestyle) interactions. PMID- 15117650 TI - [The novel non-A, non-E hepatitis viruses and their pathogenic effect]. AB - Molecular techniques have allowed the identification of new viruses in a number of patients with cryptogenic hepatitis. Whether they are clinically inapparent or true hepatitis agents remains unknown for some of them. Latest described viruses include GBV, TTV and SENV. However, based on the limited data available, they do not seem to be contenders for the new hepatitis virus title. However, researchers are looking for a role of these viruses in other chronic and acute human diseases. Only a careful evaluation of the data and the scientific concordance of all the evidence will resolve the question of whether they are only commensal viruses or pose a real pathogenic potential. PMID- 15117651 TI - [Low participation rate in breast cancer prevention initial screening: possible contributing factors]. PMID- 15117652 TI - [Prevalence of thrombotic polymorphisms in the healthy population in Southern Spain]. PMID- 15117653 TI - [Clinical outcome of beta blocker therapy in a patient with long QT syndrome with a critical mutation in the pore of potassium channel]. PMID- 15117654 TI - [Polymerase chain reaction and immunohistochemistry usefulness in refractory sprue and enteropathy-associated T cell lymphoma]. PMID- 15117655 TI - [Quality of clinical reports in emergency care]. PMID- 15117656 TI - [About cardiovascular risk assessment in the new clinical trials with statins]. PMID- 15117657 TI - Kinetic attraction during bimanual coordination. AB - Two experiments examined the effects of independent variations in kinetic and kinematic requirements on interlimb coupling during a bimanual task. The goal of the investigation was to provide preliminary evidence regarding one general class of physical variables that constrains discrete bimanual movements. Subjects attempted to execute a smooth unidirectional movement with the left arm, along with a three-segment reversal movement with the right arm. The first experiment manipulated the torque required to produce the reversal action, while movement duration and average angular velocity were held constant for both limbs. Several indications of increased interlimb coupling, due to the kinetic variation, were evident. The converse manipulation was used in the second experiment, with movement time and kinematics (velocity, acceleration) changed independently of joint torque requirements for the reversal limb. No clear effect of kinematics on coupling strength was noted. The results suggest that one variable influencing interlimb attraction toward common spatiotemporal trajectories may be kinetic in nature. PMID- 15117658 TI - Practice and the dynamics of handwriting performance: evidence for a shift of motor programming load. AB - Sixteen adult subjects were served in an experiment in which the writing of six unfamiliar graphemes was practiced. To investigate the learning process, we analyzed the absolute and relative changes of movement time of the first three consecutive segments as a function of practice. The results showed that movement time of all three segments decreased. This decrease was significantly less in the first segment that it was in the second and third segment, however. We interpret these effects of practice, from an information-processing viewpoint, as follows: (a) Initially separate response segments become integrated in more comprehensive response chunks, and (b) the preparation of later segments of the grapheme is realized more and more during the real-time execution of the initial segment. The results further revealed that these learning effects were more pronounced in graphemes composed of familiar segments than in graphemes that contained unfamiliar segments. Finally, it turned out that similarity between initial and final segments hindered the writing speed of the first segment; the effect of similarity was independent of the above-mentioned effects of practice. The latter effect is interpreted as confirming evidence for the view that the preparation of later segments of a grapheme is reflected by changes of movement time of the first segments of a grapheme. PMID- 15117659 TI - Understanding movement control in infants through the analysis of limb intersegmental dynamics. AB - One important component in the understanding of the control of limb movements is the way in which the central nervous system accounts for joint forces and torques that may be generated not only by muscle actions but by gravity and by passive reactions related to the movements of limb segments. In this study, we asked how the neuromotor system of young infants controls a range of active and passive forces to produce a stereotypic, nonintentional movement. We specifically analyzed limb intersegmental dynamics in spontaneous, cyclic leg movements (kicking) of varying intensity in supine 3-month-old human infants. Using inverse dynamics, we calculated the contributions of active (muscular) and passive (motion-dependent and gravitational) torque components at the hip, knee, and ankle joints from three-dimensional limb kinematics. To calculate joint torques, accurate estimates were needed of the limb's anthropometric parameters, which we determined using a model of the human body. Our analysis of limb intersegmental dynamics explicitly quantified the complex interplay of active and passive forces producing the simple, involuntary kicking movements commonly seen in 3-month-old infants. our results revealed that in nonvigorous kicks, hip joint reversal was the result of an extensor torque due to gravity, opposed by the combined flexor effect of the muscle torque and the total motion-dependent torque. The total motion-dependent torque increased as a hip flexor torque in more vigorous kicks; an extensor muscle torque was necessary to counteract the flexor influences of the total motion-dependent torque and, in the case of large ranges of motion, a flexor gravity torque as well. Thus, with changing passive torque influences due to motions of the linked segments, the muscle torques were adjusted to produce a net torque to reverse the kicking motion. As a consequence, despite considerable heterogeneity in the intensity, range of motion, coordination, and movement context of each kick, smooth trajectories resulted from the muscle torque, counteracting and complementing not only gravity but also the motion-dependent torques generated by movement of the linked segments. PMID- 15117660 TI - Temporal regularity of tapping by the left and right hands in timed and untimed finger tapping. AB - The temporal characteristics of repetitive finger tapping by the left and right hands were examined in two experiments. In the first experiment, interresponse intervals (IRIs) were recorded while right-handed male subjects tapped in synchrony with an auditory timing pulse (the synchronization phase) and then attempted to maintain the same tapping rate without the timing pulses (the continuation phase). The left and right hands performed separately, at four different rates (interpulse intervals of 250, 500, 750, and 1500 ms). There was no asymmetry of the asynchronies of the timing pulses and the associated responses in the synchronization phase or of the IRIs in either phase, but there was an asymmetry of chronization phase or of the IRIs in either phase, but there was an asymmetry in the temporal dispersion of the responses in both phases. in the second experiment, right-handed males tapped separately with each hand at three different speeds: as quickly as possible, at a fast but steady rate, and at a slow rhythmical rate. The speed asymmetry present when tapping as quickly as possible (with the preferred hand tapping more quickly ) was reduced when tapping at the fast steady rate and was absent when tapping at the slow rhythmical rate. The temporal dispersion of the IRIs produced by the nonpreferred hand was greater than the temporal dispersion of those produced by the preferred hand in all speed conditions. These results show smaller temporal dispersion of tapping by the preferred hand in right-handed males under different conditions, including submaximal speeds at which both hands respond at the same rate. This suggests that the motor system controlling the preferred hand in right-handers had more precise timing of response output than that controlling the nonpreferred hand. PMID- 15117661 TI - The interaction of criterion and feedback information in learning a drawing task. AB - Experiments were designed to examine the influence of criterion and feedback information in the learning of a two-dimensional drawing task. Experiment 1 showed that when the task criterion is well known to the subject, the combined presentation of criterion information and information feedback facilitates the rate of acquisition of the skill but not its overall performance level of achievement. Experiment 2 showed that when the task criterion information is not well known to the subject, presentation of criterion information facilitates both the rate of acquisition and the overall performance level and, furthermore, is essential if configuration information feedback is to be utilized effectively. Experiment 3 showed that it is the combined presentation of criterion and configuration information feedback rather than the isolate presentation of either type of information alone, that facilitates learning and performance. Collectively, the findings from the three experiments suggest an interactive effect of prior knowledge by the learner and type of augmented information in facilitating the acquisition of skill, according to the constraints imposed in the task. The data are consistent with the proposal that the degrees of freedom in the information available to support motor skill learning must match the degrees of freedom to be constraint in the perceptual-motor workspace. PMID- 15117662 TI - Motor performance of stutterers: a search for mechanisms. AB - A continuing program of research, which is concerned with identifying brain mechanisms underlying stuttering through an analysis of manual motor control, is described. Clear evidence has been found that the neural mechanisms associated with sequential responding (and, by implication, with speech) are lateralized in stutterers as they are in nonstutterers. Although no gross or general incoordination has been found in motor performance by most stutterers, their left hemisphere mechanisms appear to be inefficient for organizing and initiating new sequences of responses and vulnerable to interference from other neural activities. Results of research on bimanual coordination in stutterers are consistent with a model that attributes the interference, in part, to interhemispheric processes, possibly involving the supplementary motor area. One implication of the research is that the disfluency of stuttering is only one manifestation of a more general disfunction in motor and cognitive organization and planning. PMID- 15117663 TI - A visual representation and the control of manual aiming movements. AB - Three experiments were conducted to determine if a representation of the movement environment is functional in the organization and control of limb movements, when direct visual contact with the environment is prevented. In Experiment 1, a visual rearrangement procedure was employed to show that a representation of the environment that provides inaccurate information about the spatial location of a target can disrupt manual target aiming. In Experiment 2, we demonstrated that spatial information about the position of a target can be destroyed by a visual pattern mask, supporting our claim that the representation is visual. A target cuing procedure was used in Experiment 3 to show that representation of target position can be useful for premovement organization in a target-aiming task. Together our findings suggest that a short-lived visual representation of the movement environment may serve a useful role in the organization and control of limb movements. PMID- 15117664 TI - The effect of perceived locomotor constraints on distance estimation. AB - Two experiments were conducted to examine the ways in which the visual estimation of distance to a target is affected by constraints perceived to be placed on the subsequent locomotion to the target without vision. We hypothesized that an appraisal of impending effort would play a role in ascertaining the distance to be walked. In Experiment 1, the amount of resistance to walking was variable and unpredictable. One group of subjects performed against relatively low resistance, whereas another group performed against substantially greater resistance. In the low-resistance condition, no significant differences in CE, VE, time to target, or number of steps to target were found between any of the eight combinations of predictable or unpredictable resistances during walking. In the high-resistance condition, however, significant differences were found for CE and number of strides to target when resistance varied unpredictably during walking. Experiment 2 was similar in design but required subjects to walk with combinations of normal or short steps after they had viewed the target knowing only the gait type that would be used to begin locomotion. No differences in CE, VE, or time to target were found between four different combinations of gait type and predictability, under subjectively controlled conditions. When the step constraints were externally imposed, however, differences were found for CE. None of the results from either experiment, in which the number of strides needed to reach the target or the predictability of gait did not change from normal, supported the hypothesis that motor output requirements are necessary in forming a mental representation of the target position that can be used to walk to the target with eyes closed. Whichever locomotor technique was used to walk the estimated distance in these cases, the representation was able to be used independently. When walking mechanics were altered by externally imposed constraints, however, the success at reaching the estimated target position was reduced. These latter results are consistent with those obtained using up, down, and level walking and support the premise that mental representations used in blind walking are linked to the locomotor mechanics afforded by environmental conditions. PMID- 15117665 TI - Effects of spatial and motor demands in handwriting. AB - Four experiments were conducted to study response programming in handwriting tasks. Twelve right-handed subjects wrote acoustically presented words and phrases, and their handwriting was digitally recorded. Changes in latency, movement time, trajectory length, and pen pressure were studied as a function of response complexity (i.e., word length, complexity of initial letter, and spacing distance). The lengthening of the spatiotemporal parameters preceding the more complex structures is interpreted to be a reflection of the effects of mental load. The results further indicate that the choice of a programming strategy is dependent on the structural complexity of the task. Writing pressure decreased as a function of increased sequence length. The findings support a hierarchical model of handwriting. PMID- 15117666 TI - Arm-tracking performance with and without visual feedback in children and adults: developmental changes. AB - Tracking performance was investigated in children (aged 6-7 and 10-11) and in adult subjects. Target signals, moving unpredictably along a straight line, were tracked with the preferred arm, alternately with and without visual feedback. Qualitative observations indicate that tracking is based on continuous adjustments of the ongoing response to the continuously changing target position. No step-and-hold strategy could be detected in any of the three age groups. Tracking performance was described with four simple parameters, derived from linear systems analysis: (a) the delay between target signal and tracking movement (DL); (b) performance at the low-frequency range (LF), (c) performance at the high-frequency range (HF); and (d) a measure of tracking quality or overall similarity in the shape of target signal and tracking movement (Q). There was a considerable improvement in tracking performance with age, even after the age of 10-11, which was mainly demonstrated by a decrease in DL and increases in HF and Q. Tracking performance decreased only to a small extent when visual feedback was withdrawn. Age-related differences in the contribution of visual feedback to tracking performance could not be demonstrated. PMID- 15117668 TI - The First Conference on Motor Control in Down Syndrome. PMID- 15117667 TI - Perceptuomotor compatibility in pursuit tracking of two-dimensional movements. AB - In a previous article, we reported an investigation of visuomanual pursuit tracking of unpredictable two-dimensional targets. This article extends the study to the tracking of predictable stimuli. In both investigations, the target trajectory was elliptical. The experimental factors we varied were the orientation of the major axis of the ellipses (horizontal or vertical), the period of the movement (9.65 to 1.61 s), and the law of motion (natural vs. transformed). In the natural condition (L), the motion results from the combination of harmonic functions, as would be the case if the target were generated by a human. In the transformed (T) condition, the law of motion departs systematically from this natural model. The main results of the study are as follows: (a) Satisfactory performance is achieved only in the natural condition. Pursuit movements obey the same constraints observed in spontaneous movements. (b) Predictability affects significantly the average delay between target and pursuit. (c) Each component of the pursuit movements depends on both components of the targets. Thus, two-dimensional tracking generalizes significantly the classical one-dimensional condition. (d) The simple model developed previously to describe performance with unpredictable targets can be generalized to cover the present case as well. PMID- 15117669 TI - The effects of instructions to subjects on the programming of visually directed reaching movements. AB - Numerous studies of human motor control have examined the effects of constraints on the programming and execution of visually directed limb movements. Only a few studies, however, have explored how the subject's objective in making the movement affects the coordinated sequence of eye and limb movements that unfolds as the subject points to or grasps an object in space. In the present study, the characteristics of the targets and the environment remained constant while the demands for speed and accuracy were varied across blocks of trials by changing the instructions to the subject. In other words, the constraints operating in the situation were kept constant, but the objective of the movement was systematically varied by changing the relative demands for speed and accuracy. All subjects were required to point to visual targets presented on a screen in front of them. Eye position was monitored by infrared reflection. The position of each subject's hand in three-dimensional space was reconstructed by a computer assisted analysis of the images provided by two rotary-shutter video cameras. The speed and accuracy demands of the task were varied in blocks of trials by requiring the subjects to point to the target "as quickly as you can" (speed condition); "as accurately as you can" (accuracy condition); or both "quickly and accurately" (speed/accuracy condition). The time to initiate an eye movement to the target was found to be reduced by increasing either the speed or accuracy demands of the task although the time to initiate the hand movement was reduced only in the speed condition. While the duration of the acceleration phase of the reach remained constant in real time, the duration of the deceleration phase was increased with increased demands for accuracy. As expected, both variable and absolute errors were largest in the speed condition. The findings indicated that the programming of the limb movement and its coordination with the associated eye movements were affected by varying the objective of the task. PMID- 15117670 TI - Analysis of the transition from upright stance to steady state locomotion in children with under 200 days of autonomous walking. AB - The aim of this paper was to study, from a developmental perspective, the transient phase of gait during the period between the standing posture and the achievement of steady state gait, using temporal and biochemical parameters. Eight children who had been walking autonomously for 90 to 200 days were observed. A total of 64 sequences of steps were analyzed. A sequence of steps began with the child standing still and was executed on a large force plate. From the determination of the instantaneous velocity of center of gravity results establish that, unlike adults, progression velocity in children end of the first step, but after two to four steps. The gait initiation process does not depend on the steady state velocity, but results from an initial fall. The duration of the movement up to the end of the first step is independent of the progression velocity but depends only upon the body mass and moment of inertia of the children. PMID- 15117671 TI - Manual asymmetries: feedback processing, output variability, and spatial complexity-resolving some inconsistencies. AB - There is considerable evidence to document the case that the preferred hand is demonstrably superior for a number of manual tasks, although the mechanisms underlying this effect are less clear. Two perspectives have been dominant; one that emphasizes differential efficiency of feedback processing and one that suggests that asymmetries are a function of increased variability of output for the nonpreferred hand. This review considers the mediating effect of the complexity of the visual space in which aimed movements occur. Some inconsistencies may be resolved by noting the superiority of the right cerebral hemisphere for manipulation of spatial relationships. A multilevel, transactional perspective, which must then be adopted, may accommodate both feedback processing and motor output variability. PMID- 15117672 TI - Training for transfer of a movement timing skill. AB - Previous findings by Langley and Zelaznik (1984) suggested two hypotheses why segmental (phasing) timing training produced a more superior transfer than nonsegmental (duration) timing training. One view (the higher order variable hypothesis) suggested that segmental training developed a timing skill that was flexible for various types of transfer tasks. Another view (the contextual interference hypothesis) was that the difficulty associated with segmental training was sufficient to provide this flexibility for later transfer. The present study contrasted these hypotheses by comparing transfer following phasing or duration training but which was low in contextual interference. The acquisition results favor a contextual interference explanation. The transfer results, however, are clearly a function of the development of a higher order timing skill. These findings are discussed in terms of the development of a timing skill that is best suited for flexibility of transfer. PMID- 15117673 TI - Effects of moment of inertia on simple reaction time. AB - Two experiments examined the effect of altering the moment of inertia within an anatomical unit on simple reaction time (SRT), premotor time (PMT), and motor time (MOT) during the initiation of a discrete rapid movement. In Experiment 1 (N = 14), moment of inertia of the forearm was increased with the addition of a weighted cuff fastened around the wrist. In Experiment 2 (N = 7), moment of inertia was altered by the addition of a weighted sleeve to the index finger prior to rapid extension of the digit. Results from both experiments were unequivocal. An increase in the moment of inertia resulted in a significant increase in SRT and MOT but had no significant effect on PMT. Within selected anatomical unites (forearm and index finger), an increase in the moment of inertia does not appear to require additional neuromotor programming time but does influence the overall duration of response initiation. PMID- 15117674 TI - Discrete visual samples may control locomotor equilibrium and foot positioning in man. AB - The static or dynamic visual cues required for equilibrium as well as for foot guidance in visually guided locomotion in man were studied using a variety of locomotion supports and illumination and visual conditions. Stroboscopic illumination (brief flashes) and intermittent lighting (longer flashes) were used to control and to vary the visual sampling frequency of static (positional/orientational) visual cues. There were three main findings: First, visual control of foot positioning during locomotion over a narrow support depends mainly upon the availability of high frequency static visual cues (up to about 12 Hz); and third, static visual cues required for equilibrium control are extracted from both the peripheral and the central visual field. Assuming that discrete demands for feedback occur, a simple probabilistic model was proposed, according to which the mean time that elapses following presentation of static visual cues about positions or changes of position accounts for the differences in the difficulty of the various illumination conditions. PMID- 15117675 TI - On task and theory specificity. AB - One of the significant limitations of the motor control and skill acquisition domain is that the theories, models, and hypotheses are, in most cases, task specific. Many lines of theorizing fail to hold up under even small changes in task constraints, although clearly the field does have some robust phenomena. It is proposed that a broader consideration of the role of task constraints, which is grounded in the methodology of nonlinear dynamics, may help to formulate a more general action theory of coordination and control. PMID- 15117676 TI - Muscularis inflammation and the loss of interstitial cells of Cajal in the endothelin ETB receptor null rat. AB - Endothelin receptor null rats [ETB(-/-)] are a model for long-segment Hirschsprung's disease. These animals have significant intestinal distension (megaileum) proximal to a constricted region of the gastrointestinal tract lacking enteric ganglia. Experiments were performed to determine the pathophysiological changes that occur in these animals and to examine the tunica muscularis as a unique, immunologically active compartment. We observed abnormal intestinal flora in ETB(-/-) rats, which included a marked increase in gram negative aerobes (Enterobacteriaceae) and anaerobes (Bacteroidaceae) in the distended region of the small intestine. Histochemical observations showed that neutrophilic infiltration was rarely or not observed, but the number of ED2 positive macrophages was increased in the tunica muscularis. Expression of IL 1beta and IL-6 mRNA was also significantly increased, and the level of CD14 (LPS receptors) were increased significantly in the tunica muscularis. Spontaneous phasic contractions were irregular in the distended intestinal regions of ETB(-/ ) rats, and this was associated with an increased number of macrophages and damage to interstitial cells of Cajal (ICC) as revealed by using Kit-like immunoreactivity and electron microscopy. These results suggest that ED2-positive resident macrophages may play an important role in the inflammation of tunica muscularis in ETB(-/-) rats. Increased numbers and activation of macrophages may result in damage to ICC networks leading to disordered intestinal rhythmicity in regions of the gut in which myenteric ganglia are intact. PMID- 15117677 TI - PKC-delta and -epsilon regulate NF-kappaB activation induced by cholecystokinin and TNF-alpha in pancreatic acinar cells. AB - Although NF-kappaB plays an important role in pancreatitis, mechanisms underlying its activation remain unclear. We investigated the signaling pathways mediating NF-kappaB activation in pancreatic acinar cells induced by high-dose cholecystokinin-8 (CCK-8), which causes pancreatitis in rodent models, and TNF alpha, which contributes to inflammatory responses of pancreatitis, especially the role of PKC isoforms. We determined subcellular distribution and kinase activities of PKC isoforms and NF-kappaB activation in dispersed rat pancreatic acini. We applied isoform-specific, cell-permeable peptide inhibitors to assess the role of individual PKC isoforms in NF-kappaB activation. Both CCK-8 and TNF alpha activated the novel isoforms PKC-delta and -epsilon and the atypical isoform PKC-zeta but not the conventional isoform PKC-alpha. Inhibition of the novel PKC isoforms but not the conventional or the atypical isoform resulted in the prevention of NF-kappaB activation induced by CCK-8 and TNF-alpha. NF-kappaB activation by CCK-8 and TNF-alpha required translocation but not tyrosine phosphorylation of PKC-delta. Activation of PKC-delta, PKC-epsilon, and NF-kappaB with CCK-8 involved both phosphatidylinositol-specific PLC and phosphatidylcholine (PC)-specific PLC, whereas with TNF-alpha they only required PC-specific PLC for activation. Results indicate that CCK-8 and TNF-alpha initiate NF-kappaB activation by different PLC pathways that converge at the novel PKCs (delta and epsilon) to mediate NF-kappaB activation in pancreatic acinar cells. These findings suggest a key role for the novel PKCs in pancreatitis. PMID- 15117678 TI - Inflammatory gene expression by human colonic smooth muscle cells. AB - Intestinal mucosal cells and invading leukocytes produce inappropriate levels of cytokines and chemokines in human colitis. However, smooth muscle cells of the airway and vasculature also synthesize cytokines and chemokines. To determine whether human colonic myocytes can synthesize proinflammatory mediators, strips of circular smooth muscle and smooth muscle cells were isolated from human colon. Myocytes and muscle strips were stimulated with 10 ng/ml of IL-1beta, TNF-alpha, and IFN-gamma, respectively. Expression of mRNA for IL-1beta, IL-6, IL-8, and cyclooxygenase-2 (COX-2) was induced within 2 h and continued to increase for 8 12 h. Regulated on activation, normal T cell-expressed and -secreted (RANTES) mRNA expression was slower, appearing at 8 h and increasing linearly through 20 h. Expression of all five mRNAs was inhibited by 0.1 microM MG-132, a proteosome inhibitor that blocks NF-kappaB activation. Expression of IL-1beta, IL-6, IL-8, and COX-2 mRNA was reduced by 30 microM PP1, an Src family tyrosine kinase inhibitor, and by 25 microM SB-203580, a p38 MAPK inhibitor. MAPK/extracellular regulated kinase-1 inhibitor PD-98059 (25 microM) was much less effective. In conclusion, human colonic smooth muscle cells can synthesize and secrete interleukins (IL-1beta and IL-6) and chemokines (IL-8 and RANTES) and upregulate expression of COX-2. Regulation of cytokine, chemokine, and COX-2 mRNA depends on multiple signaling pathways, including Src-family kinases, extracellular regulated kinase, p38 MAPKs, and NF-kappaB. SB-203580 was a consistent, efficacious inhibitor of inflammatory gene expression, suggesting an important role of p38 MAPK in synthetic functions of human colonic smooth muscle. PMID- 15117679 TI - Feeding activates protein synthesis in mouse pancreas at the translational level without increase in mRNA. AB - To determine the mechanism of meal-regulated synthesis of pancreatic digestive enzymes, we studied the effect of fasting and refeeding on pancreatic protein synthesis, relative mRNA levels of digestive enzymes, and activation of the translational machinery. With the use of the flooding dose technique with L [3H]phenylalanine, morning protein synthesis in the pancreas of Institute for Cancer Research mice fed ad libitum was 7.9 +/- 0.3 nmol phenylalanine.10 min( 1).mg protein(-1). Prior fasting for 18 h reduced total protein synthesis to 70 +/- 1.4% of this value. Refeeding for 2 h, during which the mice consumed 29% of their daily food intake, increased protein synthesis to 117.3 +/- 4.9% of the control level. Pancreatic mRNA levels of amylase, lipases, trypsins, chymotrypsin, elastases, as well as those for several housekeeping genes tested were not significantly changed after refeeding compared with fasted mice. By contrast, the major translational control pathway involving Akt, mTOR, and S6K was strongly regulated by fasting and refeeding. Fasting for 18 h decreased phosphorylation of ribosomal protein S6 to almost undetectable levels, and refeeding highly increased it. The most highly phosphorylated form of the eIF4E binding protein (4E-BP1) made up the 14.6% of total 4E-BP1 in normally fed animals, was only 2.8% after fasting, and was increased to 21.4% after refeeding. This was correlated with an increase in the formation of the eIF4E-eIF4G complex after refeeding. By contrast, feeding did not affect eIF2B activity. Thus food intake stimulates pancreatic protein synthesis and translational effectors without increasing digestive enzyme mRNA levels. PMID- 15117680 TI - Tobacco sales to minors: has familiarity bred contempt for youth access programs? PMID- 15117681 TI - Oral health care for the elderly: more than just dentures. PMID- 15117682 TI - Oral health care services for older adults: a looming crisis. PMID- 15117683 TI - Strengthening the oral health safety net: delivery models that improve access to oral health care for uninsured and underserved populations. PMID- 15117684 TI - What's new about the "new public health"? AB - From its origins, when public health was integral to societies' social structures, through the sanitary movement and contagion eras, when it evolved as a separate discipline, to the "new public health" era, when health promotion projects like Healthy Cities appear to be steering the discipline back to society's social structure, public health seems to have come full circle. It is this observation that has led some to ask, "What's new about the 'new public health'?" This article addresses the question by highlighting what is new about the health promotion era-including adapted components of previous eras that have been incorporated into its core activities-and its suitability in addressing established and emerging public health threats. PMID- 15117685 TI - Professional and hospital discrimination and the US Court of Appeals Fourth Circuit 1956-1967. AB - A series of court cases litigated by the National Association for the Advancement of Colored People Legal Defense and Education Fund between 1956 and 1967 laid the foundation for elimination of overt discrimination in hospitals and professional associations. The landmark case, Simkins v Moses H. Cone Memorial Hospital (1963), challenged the use of public funds to expand segregated hospital care. The second case, Cypress v Newport News Hospital Association (1967), reaffirmed the federal government's application of Medicare certification guidelines to force hospitals to open up patient admissions, education programs, and staff privileges to all citizens and physicians. Pursuit of a legal strategy against racist policies was an essential element in a national campaign to eliminate discrimination in health care delivery in the United States. PMID- 15117686 TI - Popularizing the toothbrush. PMID- 15117687 TI - An address on tooth culture. PMID- 15117688 TI - Sir James Crichton-Browne: Victorian psychiatrist and public health reformer. PMID- 15117689 TI - Child health: reaching the poor. AB - In most countries, rates of mortality and malnutrition among children continue to decline, but large inequalities between poor and better-off children exist, both between and within countries. These inequalities, which appear to be widening, call into question the strategies for child mortality reduction relied upon to date. We review (1) what is known about the causes of socioeconomic inequalities in child health and where programs aimed at reducing inequalities may be most effectively focused and (2) what is known about the success of actual programs in narrowing these inequalities. We end with lessons learned: the need for better evidence, but most of all for a new approach to improving the health of all children that is evidence based, broad, and multifaceted. PMID- 15117690 TI - John Henryism and self-reported physical health among high-socioeconomic status African American men. AB - We performed a cross-sectional survey of high-socioeconomic status (SES) African American men and their health to examine the relationship between John Henryism (the strong behavioral predisposition to directly confront barriers to upward social mobility) and self-reported physical health status. We found a positive association between John Henryism and better physical health among high-SES African American men. The study of social and behavioral implications of health of men of differing SES is required to develop strategies to improve the health of African American men. PMID- 15117691 TI - Swimmer's itch: incidence and risk factors. AB - Swimmer's itch (cercarial dermatitis) affects people engaged in open-water activities. We report incidence and risk factors for a US lake. Water exposures and swimmer's itch experience were reported daily for riparian household residents and guests at Douglas Lake, Michigan, in July 2000. Incidence of swimmer's itch was 6.8 episodes per 100 water exposure days. Positive risks were (1) exposures in shallow water and in areas with onshore winds and (2) more days of lake use in July. Further epidemiological studies will help public health agencies address this bothersome problem at recreational lakes. PMID- 15117692 TI - Cancer burden from arsenic in drinking water in Bangladesh. AB - We assessed the potential burden of internal cancers due to arsenic exposure in Bangladesh. We estimated excess lifetime risks of death from liver, bladder, and lung cancers using an exposure distribution, death probabilities, and cancer mortality rates from Bangladesh and dose-specific relative risk estimates from Taiwan. Results indicated at least a doubling of lifetime mortality risk from liver, bladder, and lung cancers (229.6 vs 103.5 per 100 000 population) in Bangladesh owing to arsenic in drinking water. PMID- 15117693 TI - Computer access and Internet use among urban youths. AB - This report presents data on computer access, Internet use, and factors associated with health information seeking on the Internet among a sample of youths aged 15 to 30 years in New York City. Findings from street intercept surveys indicate substantial computer access at home (62%) and frequent (everyday or a few times a week) Internet use (66%). Fifty-five percent of the sample reported seeking health information on the Internet, which was associated with positive beliefs about getting a health checkup and frequent Internet use. PMID- 15117694 TI - Prevalence of multiple chemical sensitivities: a population-based study in the southeastern United States. AB - We examined the prevalence of multiple chemical sensitivities (MCS), a hypersensitivity to common chemical substances. We used a randomly selected sample of 1582 respondents from the Atlanta, Ga, standard metropolitan statistical area. We found that 12.6% of our sample reported the hypersensitivity and that, while the hypersensitivity is more common in women, it is experienced by both men and women of a variety of ages and educational levels. Our prevalence for MCS is similar to that (15.9%) found by the California Department of Health Services in California and suggests that the national prevalence may be similar. PMID- 15117696 TI - Oral disease burden in Northern Manhattan patients with diabetes mellitus. AB - OBJECTIVES: We explored the association between diabetes mellitus and oral disease in a low-socioeconomic-status urban population. METHODS: Dental records of 150 adults with diabetes and 150 nondiabetic controls from the dental clinic at Columbia University in Northern Manhattan matched by age and gender were studied. RESULTS: There was a 50% increase in alveolar bone loss in diabetic patients compared with nondiabetic controls. Diabetes, increasing age, male gender, and use of tobacco products had a statistically significant effect on bone loss. CONCLUSIONS: Our findings provide evidence that diabetes is an added risk for oral disease in this low-income, underserved population of Northern Manhattan. Oral disease prevention and treatment programs may need to be part of the standards of continuing care for patients with diabetes PMID- 15117695 TI - Social factors and periodontitis in an older population. AB - OBJECTIVES: We assessed the prevalences of periodontitis by education and income levels among US adults with data from the third National Health and Nutrition Examination Survey. METHODS: The study was limited to non-Hispanic Blacks, Mexican Americans, and non-Hispanic Whites 50 years of age or older with a complete periodontal assessment during the dental examination. RESULTS: Blacks with higher education and income levels had a significantly higher prevalence of periodontitis than their White and Mexican-American counterparts. The relationship between income level and periodontitis was modified by race/ethnicity. High-income Blacks exhibited a higher prevalence of periodontitis than did low-income Blacks and high-income Whites. CONCLUSIONS: Our findings call attention to the importance of recognizing socioeconomic status-related health differences across racial/ethnic groups within the social, political, and historical context. PMID- 15117697 TI - Dental insurance visits and expenditures among older adults. AB - OBJECTIVES: We examined the effect of age, income, and coverage on dental service utilization during 1996. METHODS: We used data from the 1996 Medical Expenditure Panel Survey. RESULTS: Edentulous and poorer older adults are less likely to have coverage and less likely to report a dental visit than dentate or wealthier older adults. CONCLUSIONS: These analyses help to describe the needs of older adults as they cope with diminishing resources as a consequence of retirement, including persons previously accustomed to accessing oral health services with dental insurance. PMID- 15117698 TI - Dental care use and self-reported dental problems in relation to pregnancy. AB - OBJECTIVES: We examined the relationships between risk factors amenable to intervention and the likelihood of dental care use during pregnancy. METHODS: We used data from the Washington State Department of Health's Pregnancy Risk Assessment Monitoring System. RESULTS: Of the women surveyed, 58% reported no dental care during their pregnancy. Among women with no dental problems, those not receiving dental care were at markedly increased risk of having received no counseling on oral health care, being overweight, and using tobacco. Among women who received dental care, those with dental problems were more likely to have lower incomes and Medicaid coverage than those without dental problems. CONCLUSIONS: There is a need for enhanced education and training of maternity care providers concerning oral health in pregnancy. PMID- 15117699 TI - Effects of WIC participation on children's use of oral health services. AB - OBJECTIVES: We estimated the effects of the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) on dental services use by Medicaid children in North Carolina. METHODS: We used linked Medicaid claims and enrollment files, WIC files, and the area resource file to compare dental services use for children enrolled in WIC with those not enrolled. We used multivariate models that controlled for child clustering and employed 2-step methodology to control for selection bias. RESULTS: Children who participated in WIC had an increased probability of having a dental visit, were more likely to use preventive and restorative services, and were less likely to use emergency services. CONCLUSIONS: Children's WIC participation improved access to dental care services that should lead to improved oral health. PMID- 15117700 TI - Does supplemental private insurance affect care of Medicare recipients hospitalized for myocardial infarction? AB - OBJECTIVES: We sought to determine whether supplemental private insurance coverage among Medicare recipients alters patterns of health care or outcomes associated with acute myocardial infarction. METHODS: Medicare patients hospitalized after a myocardial infarction were identified from New York City hospitalization records. Patients who had only Medicare coverage were compared with those who had supplemental private or public insurance coverage. RESULTS: Patients with supplemental private insurance exhibited increased rates of revascularization and decreased rates of in-hospital mortality relative to patients with either Medicare only or Medicare and public insurance. Moreover, Blacks and women were less likely to undergo revascularization and exhibited higher in-hospital mortality rates. CONCLUSIONS: Despite Medicare, private insurance coverage appears to influence the likelihood of coronary revascularization among older patients hospitalized for acute myocardial infarction. PMID- 15117701 TI - Delays and unmet need for health care among adult primary care patients in a restructured urban public health system. AB - OBJECTIVES: We estimated the prevalence and determinants of delayed and unmet needs for medical care among patients in a restructured public health system. METHODS: We conducted a stratified cross-sectional probability sample of primary care patients in the Los Angeles County Department of Health Services. Face-to face interviews were conducted with 1819 adult patients in 6 languages. The response rate was 80%. The study sample was racially/ethnically diverse. RESULTS: Thirty-three percent reported delaying needed medical care during the preceding 12 months; 25% reported an unmet need for care because of competing priorities; and 46% had either delayed or gone without care. CONCLUSIONS: Barriers to needed health care continue to exist among patients receiving care through a large safety net system. Competing priorities for basic necessities and lack of insurance contribute importantly to unmet health care needs. PMID- 15117702 TI - Assessment of household food security among food stamp recipient families in Maryland. AB - OBJECTIVES: We assessed the relationship between food security status and various sociodemographic characteristics among households that include children and that receive food stamps. METHODS: A modified version of the US Food Security Survey Module was implemented by telephone survey with Maryland food stamp recipients. RESULTS: Of the 245 households, 66% experienced food insecurity. Food security status was associated with participation in the Special Supplemental Food Program for Women, Infants, and Children, the summer food program, and a food bank. Food security status was not associated with the number of months households received food stamps. There was no difference between the food security status of households living in urban and rural counties. CONCLUSIONS: A gap exists between the food stamp support provided and some households' nutritional and economic needs. PMID- 15117703 TI - Use of preventive services by men enrolled in Medicare+Choice plans. AB - OBJECTIVES: We examined the effect of demographic and socioeconomic factors on use of preventive services (prostate-specific antigen testing, colorectal cancer screening, and influenza vaccination) among elderly men enrolled in 2 Medicare+Choice health plans. METHODS: Data were derived from administrative files and a survey of 1915 male enrollees. We used multivariate logistic regression to assess the effects of enrollee characteristics on preventive service use. RESULTS: Age, marital status, educational attainment, and household wealth were associated with receipt of one or more preventive services. However, the effects of these variables were substantially attenuated relative to earlier studies of Medicare. CONCLUSIONS: Some Medicare HMOs have been successful in attenuating racial and socioeconomic disparities in the use of preventive services by older men. PMID- 15117704 TI - Racial differences in survival among men with prostate cancer and comorbidity at time of diagnosis. AB - OBJECTIVES: This study evaluated the effect of comorbidity at diagnosis on racial differences in survival among men with prostate cancer. METHODS: Clinical and demographic data were abstracted from records of 864 patients diagnosed at 4 Chicago area hospitals between 1986 and 1990. Comorbidity was scored on the basis of clinical information in the Charlson index. Cause-specific relative mortality adjusted for age, stage, differentiation, and treatment was compared across Charlson scores with Cox proportional hazards functions. RESULTS: Blacks had significantly greater mortality from prostate cancer and other causes (vs Whites, relative risk [95% confidence interval] = 1.84 [1.22, 2.79] and 1.69 [1.33, 2.29], respectively; P <.001). However, differences disappeared as initial comorbidity increased (1.75 [1.33, 2.31] vs 0.90 [0.59, 1.29] for scores = 0 and > or =5, respectively). CONCLUSIONS: Absence of a significant preexisting medical diagnosis is associated with a higher risk for excess mortality among Black men diagnosed with prostate cancer. PMID- 15117705 TI - Effect of discrimination on mental health service utilization among Chinese Americans. AB - OBJECTIVES: We examined the association between discrimination and mental health service use among a representative sample of Chinese Americans. METHODS: Our data were derived from the 2-wave Chinese American Psychiatric Epidemiological Survey, a strata-cluster survey conducted in 1993 and 1994 in a western American city. RESULTS: Language-based discrimination was associated with higher levels of use of informal services and seeking help from friends and relatives for emotional problems. Negative attitudes toward professional mental health services were associated with greater use of informal services. CONCLUSIONS: The findings suggest that language-based discrimination influences patterns of mental health service use among Chinese Americans. Implications for service providers and policymakers are discussed. PMID- 15117706 TI - Factors associated with colorectal cancer screening among the US urban Japanese population. AB - OBJECTIVES: The author examined the prevalence and predictors of colorectal cancer screening among the urban Japanese population of the United States. METHODS: A sample of Japanese residents of major US metropolitan areas completed a self-administered mailed survey. RESULTS: Physician recommendation, acculturation, and perceived psychological costs were consistent predictors of screening for colorectal cancer. Gender and marital status were related to screening via fecal occult blood testing; age, susceptibility, and health insurance were related to sigmoidoscopy/colonoscopy screening. CONCLUSIONS: Colorectal cancer screening among the urban Japanese population could be increased with interventions seeking to promote physician recommendations for screening, alleviate perceived psychological costs among patients, and improve physician-patient communication. PMID- 15117707 TI - Vision impairment and hearing loss among community-dwelling older Americans: implications for health and functioning. AB - OBJECTIVES: We investigated the health, activity, and social participation of people aged 70 years or older with vision impairment, hearing loss, or both. METHODS: We examined the 1994 Second Supplement on Aging to determine the health and activities of these 3 groups compared with those without sensory loss. We calculated odds ratios and classified variables according to the International Classification of Functioning, Disability and Health framework. RESULTS: Older people with only hearing loss reported disparities in health, activities, and social roles; those with only vision impairment reported greater disparities; and those with both reported the greatest disparities. CONCLUSIONS: A hierarchical pattern emerged as impairments predicted consistent disparities in activities and social participation. This population's patterns of health and activities have public health implications. PMID- 15117708 TI - Self-reported changes in drug and alcohol use after becoming homeless. AB - OBJECTIVES: We identified substance use patterns and factors associated with increased substance use after users become homeless. METHODS: We carried out a 2 city, community-based survey that used population-proportionate sampling of 91 sites with random selection at each site. RESULTS: Five hundred thirty-one adults were interviewed; 78.3% of them met Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition criteria for substance abuse or dependence. Most of those who met the criteria reported using drugs and alcohol less since they became homeless, commonly because they were in recovery. Factors independently associated with increased use were no health insurance (odds ratio [OR] = 1.6; 95% confidence interval [CI] = 1.02, 2.58), alcohol abuse or dependence (OR = 3.5; 95% CI = 1.85, 6.78), and selling plasma (OR = 2.6; 95% CI = 1.32, 5.14) or panhandling (OR = 3.0; 95% CI = 1.65, 5.55) to acquire drugs. CONCLUSIONS: Becoming homeless plays a role in self-reported substance use. Multiservice treatment programs and tailored interventions for homeless persons are needed. PMID- 15117709 TI - The limited relevance of drug policy: cannabis in Amsterdam and in San Francisco. AB - OBJECTIVES: We tested the premise that punishment for cannabis use deters use and thereby benefits public health. METHODS: We compared representative samples of experienced cannabis users in similar cities with opposing cannabis policies Amsterdam, the Netherlands (decriminalization), and San Francisco, Calif (criminalization). We compared age at onset, regular and maximum use, frequency and quantity of use over time, intensity and duration of intoxication, career use patterns, and other drug use. RESULTS: With the exception of higher drug use in San Francisco, we found strong similarities across both cities. We found no evidence to support claims that criminalization reduces use or that decriminalization increases use. CONCLUSIONS: Drug policies may have less impact on cannabis use than is currently thought. PMID- 15117710 TI - Draw the line/respect the line: a randomized trial of a middle school intervention to reduce sexual risk behaviors. AB - OBJECTIVES: This study evaluated the long-term effectiveness of Draw the Line/Respect the Line, a theoretically based curriculum designed to reduce sexual risk behaviors among middle school adolescents. METHODS: The randomized controlled trial involved 19 schools in northern California. A cohort of 2829 sixth graders was tracked for 36 months. RESULTS: The intervention delayed sexual initiation among boys, but not girls. Boys in the intervention condition also exhibited significantly greater knowledge than control students, perceived fewer peer norms supporting sexual intercourse, had more positive attitudes toward not having sex, had stronger sexual limits, and were less likely to be in situations that could lead to sexual behaviors. Psychosocial effects for girls were limited. CONCLUSIONS: The program was effective for boys, but not for girls. PMID- 15117711 TI - Adolescents and firearms: a California statewide survey. AB - OBJECTIVES: We assessed the prevalence and correlates of adolescents' reports regarding firearms in their homes, of their own, of close friends, and of same aged peers. METHODS: Random-digit-dialed interviews were conducted with 5801 adolescents as part of the California Health Interview Survey. RESULTS: One fifth (19.6%) of California adolescents reported having a firearm in their homes; few (3.0%) reported having their own gun. Characteristics associated with having one's own gun and with perceptions regarding others' guns generally were consistent with characteristics associated with having a firearm in the home. The 2 exceptions were related to socioeconomic status and to ethnicity. CONCLUSIONS: The source from which adolescents obtain guns, especially adolescents from less wealthy households, merits further investigation. Further research is needed to ascertain the accuracy of Black and Latino adolescents' perceptions regarding handguns among their peers. PMID- 15117712 TI - Can better mental health services reduce the risk of juvenile justice system involvement? AB - OBJECTIVES: We evaluated how improved mental health services affect justice involvement among juveniles treated in the public mental health system. METHODS: Our analyses were based on administrative and interview data collected in 2 communities participating in the evaluation of a national initiative designed to improve mental health services for children and youths. RESULTS: Results derived from Cox proportional hazard models suggested that better mental health services reduced the risks of initial and subsequent juvenile justice involvement by 31% and 28%, respectively. Effects were somewhat more pronounced for serious offenses. CONCLUSIONS: Our findings suggest that improved mental health services reduce the risk of juvenile justice involvement. PMID- 15117713 TI - Overcoming language barriers in health care: costs and benefits of interpreter services. AB - OBJECTIVES: We assessed the impact of interpreter services on the cost and the utilization of health care services among patients with limited English proficiency. METHODS: We measured the change in delivery and cost of care provided to patients enrolled in a health maintenance organization before and after interpreter services were implemented. RESULTS: Compared with English speaking patients, patients who used the interpreter services received significantly more recommended preventive services, made more office visits, and had more prescriptions written and filled. The estimated cost of providing interpreter services was $279 per person per year. CONCLUSIONS: Providing interpreter services is a financially viable method for enhancing delivery of health care to patients with limited English proficiency. PMID- 15117714 TI - Vitamin C deficiency and depletion in the United States: the Third National Health and Nutrition Examination Survey, 1988 to 1994. AB - OBJECTIVES: We sought to determine prevalence rates of vitamin C deficiency and depletion in the United States. METHODS: We used data from the Third National Health and Nutrition Examination Survey to assess intake of dietary, supplemental, and serum vitamin C. RESULTS: Mean intakes and serum levels of vitamin C were normal; however, vitamin C deficiency and depletion were common (occurring among 5%-17% and 13%-23% of respondents, respectively). Smokers, those who did not use supplements, and non-Hispanic Black males had elevated risks of vitamin C deficiency, while Mexican Americans had lower risks. CONCLUSIONS: Health professionals should recommend consumption of vegetables and fruits rich in vitamin C and should recommend supplementation for individuals at risk of vitamin C deficiency. PMID- 15117715 TI - Highway repair: a new silicosis threat. AB - OBJECTIVES: We describe an emerging public health concern regarding silicosis in the fast-growing highway repair industry. METHODS: We examined highway construction trends, silicosis surveillance case data, and environmental exposure data to evaluate the risk of silicosis among highway repair workers. We reviewed silicosis case data from the construction industry in 3 states that have silicosis registries, and we conducted environmental monitoring for silica at highway repair work sites. RESULTS: Our findings indicate that a large population of highway workers is at risk of developing silicosis from exposure to crystalline silica. CONCLUSIONS: Exposure control methods, medical screenings, protective health standards, and safety-related contract language are necessary for preventing future occupational disease problems among highway repair workers. PMID- 15117716 TI - Comparison of Ca2+ release and uptake characteristics of the sarcoplasmic reticulum in isolated horse and rabbit cardiomyocytes. AB - Both the cardiac action potential duration (APD) (0.6-1 s) and resting heart rate (30-40 beats/min) in the horse are significantly different from humans and smaller mammals, including the rabbit. This would be anticipated to have consequences for excitation-contraction (EC) coupling and require adaptation of the individual processes involved. The sarcoplasmic reticulum (SR) is one of the main components involved in EC coupling. This study examines and compares the activity of this organelle in the horse with that of the rabbit. In particular, the study focuses on SR Ca2+ release via the Ca2+ release channel/ryanodine receptor (RyR2) and Ca2+ uptake via the sarco(endo)plasmic reticulum Ca(2+) ATPase (SERCA) pump. Isolated cardiomyocytes from both horse and rabbit hearts were permeabilized, bathed in a mock intracellular solution, and exposed to a specified [Ca2+]. Rabbit cardiomyocytes exposed to 260 nM [Ca2+] produced spontaneous Ca2+ release and propagated Ca2+ waves. Horse cells failed to produce Ca2+ waves; instead, only local release in the form of Ca2+ sparks was evident. However, at 550 nM [Ca2+], Ca2+ waves were produced in both species. Ca2+ waves were four times less frequent yet approximately 1.5 times greater in amplitude in the horse compared with the rabbit. Ca2+ wave velocity was comparable between the species. The reason for this disparity in Ca2+ wave characteristics is unknown. Separate measurements of oxalate-supported Ca2+ uptake into the SR suggest that both horse and rabbit cardiomyocytes have comparable levels SERCA activity. The possible reasons for the observed differences in SR Ca2+ release between the horse and rabbit are discussed. PMID- 15117717 TI - Regional blood volume and peripheral blood flow in postural tachycardia syndrome. AB - Variants of postural tachycardia syndrome (POTS) are associated with increased ["high-flow" POTS (HFP)], decreased ["low-flow" POTS (LFP)], and normal ["normal flow" POTS (NFP)] blood flow measured in the lower extremities while subjects were in the supine position. We propose that postural tachycardia is related to thoracic hypovolemia during orthostasis but that the patterns of peripheral blood flow relate to different mechanisms for thoracic hypovolemia. We studied 37 POTS patients aged 14-21 yr: 14 LFP, 15 NFP, and 8 HFP patients and 12 healthy control subjects. Peripheral blood flow was measured in the supine position by venous occlusion strain-gauge plethysmography of the forearm and calf to subgroup patients. Using indocyanine green techniques, we showed decreased cardiac index (CI) and increased total peripheral resistance (TPR) in LFP, increased CI and decreased TPR in HFP, and unchanged CI and TPR in NFP while subjects were supine compared with control subjects. Blood volume tended to be decreased in LFP compared with control subjects. We used impedance plethysmography to assess regional blood volume redistribution during upright tilt. Thoracic blood volume decreased, whereas splanchnic, pelvic, and leg blood volumes increased, for all subjects during orthostasis but were markedly lower than control for all POTS groups. Splanchnic volume was increased in NFP and LFP. Pelvic blood volume was increased in HFP only. Calf volume was increased above control in HFP and LFP. The results support the hypothesis of (at least) three pathophysiologic variants of POTS distinguished by peripheral blood flow related to characteristic changes in regional circulations. The data demonstrate enhanced thoracic hypovolemia during upright tilt and confirm that POTS is related to inadequate cardiac venous return during orthostasis. PMID- 15117718 TI - High-resolution imaging reveals a limit in spatial resolution of blood flow measurements by microspheres. AB - Density of 15-microm microspheres after left atrial application is the standard measure of regional perfusion. In the heart, substantial differences in microsphere density are seen at spatial resolutions <5 ml, implying perfusion heterogeneity. Microsphere deposition imaging permits a superior evaluation of the distribution pattern. Therefore, fluorescent microspheres (FMS) were applied, FMS deposition in the canine heart was imaged by epifluorescence microscopy in vitro, and the patterns were observed compared with MR images of iron oxide microspheres (IMS) obtained in vivo and in vitro. FMS deposition in myocardial slices revealed the following: 1) a nonrandom distribution, with sequentially applied FMS of different color stacked within the same vessel, 2) general FMS clustering, and 3) rather large areas devoid of FMS (n = 3). This pattern was also seen in reconstructed three-dimensional images (<1 nl resolution) of FMS distribution (n = 4). Surprisingly, the deposition pattern of sequentially applied FMS remained virtually identical over 3 days. Augmenting flow by intracoronary adenosine (>2 microM) enhanced local microsphere density, but did not alter the deposition pattern (n = 3). The nonrandom, temporally stable pattern was quantitatively confirmed by a three-dimensional intermicrosphere distance analysis of sequentially applied FMS. T2-weighted short-axis MR images (2-microl resolution) of IMS revealed similar patterns in vivo and in vitro (n = 6), as seen with FMS. The observed temporally stable microsphere patterns are not consistent with the notion that microsphere deposition is solely governed by blood flow. We propose that at high spatial resolution (<2 microl) structural aspects of the vascular network dominate microsphere distribution, resulting in the organized patterns observed. PMID- 15117719 TI - Influence of structural geometry on the severity of bicuspid aortic stenosis. AB - Doppler-derived gradients may overestimate total pressure loss in degenerative and prosthetic aortic valve stenosis (AS) due to unaccounted pressure recovery distal to the orifice. However, in congenitally bicuspid valves, jet eccentricity may result in a higher anatomic-to-effective orifice contraction ratio, resulting in an increased pressure loss at the valve and a reduced pressure recovery distal to the orifice leading to greater functional severity. The objective of our study was to determine the impact of local geometry on the total versus Doppler-derived pressure loss and therefore the assessed severity of the stenosis in bicuspid valves. On the basis of clinically obtained measurements, two- and three dimensional computer simulations were created with various local geometries by altering the diameters of the left ventricular outflow tract (LVOT; 1.8-3.0 cm), orifice diameter (OD; 0.8-1.6 cm), and aortic root diameter (AR; 3.0-5.4 cm). Jet eccentricity was altered in the models from 0 to 25 degrees. Simulations were performed under steady-flow conditions. Axisymmetric simulations indicate that the overall differences in pressure recovery were minor for variations in LVOT diameter (<3%). However, both OD and AR had a significant impact on pressure recovery (6-20%), with greatest recovery being the larger OD and the smaller recovery being the AR. In addition, three-dimensional data illustrate a greater pressure loss for eccentric jets with the same orifice area, thus increasing functional severity. In conclusion, jet eccentricity results in greater pressure loss in bicuspid valve AS due to reduced effective orifice area. Functional severity may also be enhanced by larger aortic roots, commonly occurring in these patients, leading to reduced pressure recovery. Thus, for the same anatomic orifice area, functional severity is greater in bicuspid than in degenerative tricuspid AS. PMID- 15117720 TI - Radicicol activates heat shock protein expression and cardioprotection in neonatal rat cardiomyocytes. AB - Heat shock proteins (HSPs) constitute an endogenous cellular defense mechanism against environmental stresses. In the past few years, studies have shown that overexpression of HSPs can protect cardiac myocytes against ischemia-reperfusion injury. In an attempt to increase the HSPs in cardiac tissue, we used the compound radicicol that activates HSP expression by binding to the HSP 90 kDa (HSP90). HSP90 is the main component of the cytosolic molecular chaperone complex, which has been implicated in the regulation of the heat shock factor 1 (HSF1). HSF1 is responsible for the transcriptional activation of the heat shock genes. In the present study, we show that radicicol induces HSP expression in neonatal rat cardiomyocytes, and this increase in HSPs confers cardioprotection to these cardiomyocytes. We also show that radicicol induction of the HSP and cardioprotection is dependent on the inhibition of HSP90 in cardiomyocytes. These results indicate that modulation of the active HSP90 protein level plays an important role in cardioprotection. Therefore, compounds, such as radicicol and its possible derivatives that inhibit the function of HSP90 in the cell may represent potentially useful cardioprotective agents. PMID- 15117721 TI - Effect of neonatal dexamethasone exposure on growth and neurological development in the adult rat. AB - Until recently, the synthetic glucocorticoid dexamethasone was commonly used to lessen the morbidity of chronic lung disease in premature infants. This practice diminished as dexamethasone use was linked to an increased incidence of cerebral palsy and short-term neurodevelopmental delay. Of more concern is the fact that we know little regarding dexamethasone effects on long-term neurodevelopment. To study the effects of neonatal dexamethasone exposure on long-term neurodevelopment, we have developed a rat model where newborn pups are exposed to tapering doses of dexamethasone at time points corresponding to the neurodevelopmental age when human infants are traditionally exposed to this drug in the neonatal intensive care unit. Using a within-litter design, pups were assigned to one of three groups on postnatal day 2 (P2): handled controls, saline injected controls, and animals receiving intramuscular dexamethasone between P3 and P6. Somatic growth was decreased in dexamethasone-treated animals. Dexamethasone-treated animals demonstrated slight delays in indexes of neurodevelopment and physical maturation at P7 and P14, but not P20. In adolescence (P45), there was no difference between groups in an open field test. However, as adult dexamethasone-treated animals were less active in the open field and spent more time in closed arms of the elevated plus maze. The serum corticosterone response to crowding stress in dexamethasone-treated animals was no different from controls, but they demonstrate a delay in return of corticosterone levels to baseline. These differences in behavior and hormonal stress responsiveness suggest that neonatal dexamethasone exposure may permanently alter function of the neuroendocrine stress axis. PMID- 15117722 TI - Effect of acute exercise and exercise training on VEGF splice variants in human skeletal muscle. AB - The present study investigated the effect of an acute exercise bout on the mRNA response of vascular endothelial growth factor (VEGF) splice variants in untrained and trained human skeletal muscle. Seven habitually active young men performed one-legged knee-extensor exercise training at an intensity corresponding to approximately 70% of the maximal workload in an incremental test five times/week for 4 wk. Biopsies were obtained from the vastus lateralis muscle of the trained and untrained leg 40 h after the last training session. The subjects then performed 3 h of two-legged knee-extensor exercise, and biopsies were obtained from both legs after 0, 2, 6, and 24 h of recovery. Real-time PCR was used to examine the expression of VEGF mRNA containing exon 1 and 2 (all VEGF isoforms), exon 6 or exon 7, and VEGF(165) mRNA. Acute exercise induced an increase (P < 0.05) in total VEGF mRNA levels as well as VEGF(165) and VEGF splice variants containing exon 7 at 0, 2, and 6 h of recovery. The increase in VEGF mRNA was higher in the untrained than in the trained leg (P < 0.05). The results suggest that in human skeletal muscle, acute exercise increases total VEGF mRNA, an increase that appears to be explained mainly by an increase in VEGF(165) mRNA. Furthermore, 4 wk of training attenuated the exercise-induced response in skeletal muscle VEGF(165) mRNA. PMID- 15117723 TI - Salutary effects of androstenediol on cardiac function and splanchnic perfusion after trauma-hemorrhage. AB - Recent studies have shown that dehydroepiandrosterone (DHEA) administration after trauma-hemorrhage (T-H) improves cardiovascular function and decreases cytokine production in male animals. Although androstenediol, one of the metabolites of DHEA, is reported to have estrogen-like activity, it remains unknown whether androstenediol per se has any salutary effects on cytokines and cardiovascular function after T-H. To examine this effect, male Sprague-Dawley rats underwent laparotomy and were bled to and maintained at a mean arterial blood pressure of 35-40 mmHg for approximately 90 min. The animals were resuscitated with four times the volume of maximal bleedout volume in the form of Ringer lactate. Androstenediol (1 mg/kg body wt i.v.) or vehicle was administered at the end of resuscitation. Twenty-four hours after resuscitation, cardiac function and organ blood flow were measured by using (85)Sr-microspheres. Circulating levels of nitrate/nitrite and IL-6 were also determined. Cardiovascular function and organ blood flow were significantly depressed after T-H. However, these parameters were restored by androstenediol treatment. The elevated plasma IL-6 levels after T-H were also lowered by androstenediol treatment. In contrast, plasma levels of nitrate/nitrite were the highest in the androstenediol-treated T-H animals. Because androstenediol administration after T-H decreases cytokine production and improves cardiovascular function, this agent appears to be a novel and useful adjunct for restoring the depressed cardiovascular function and for cytokine production in males after adverse circulatory conditions. PMID- 15117724 TI - Constitutive roles for inducible genes: evidence for the alteration in expression of the inducible hsp70 gene in Antarctic notothenioid fishes. AB - Previous research on the Antarctic notothenioid fish Trematomus bernacchii demonstrated the loss of the heat shock response (HSR), a classical cellular defense mechanism against thermal stress, characterized by the rapid synthesis of heat shock proteins (Hsps). In the current study, we examined potential mechanisms for the apparent loss of the HSR in Antarctic notothenioids and, in addition, compared expression patterns of two genes from the 70-kDa Hsp family (hsc71 and hsp70) in tissues from T. bernacchii to expression patterns in tissues of two closely related temperate notothenioid fishes from New Zealand, Bovichtus variegatus and Notothenia angustata. The results showed that transcript for both the constitutive and inducible genes in the Hsp70 gene family were expressed in detectable levels in all three species. However, only the cold-temperate New Zealand fishes displayed the ability to upregulate the inducible transcript, hsp70. Although hsp70 was present in detectable levels in several tissues of the Antarctic notothen T. bernacchii, in vitro thermal stresses failed to produce a significant increase in mRNA levels. In all species, the expression of the constitutive transcript hsc71 was variable and nonresponsive to temperature increases, even at temperatures as high as 10 degrees C above the ecologically relevant range for the species under study. Field-collected tissues from T. bernacchii (sampled immediately after capture) indicated that hsp70 mRNA was expressed at high levels in field-acclimatized fishes. Thus upregulation of molecular chaperones suggested that low-temperature stress may be significantly denaturing to cellular proteins in Antarctic fish, an observation that was supported by elevated levels of ubiquitin-conjugated protein. PMID- 15117725 TI - Hypoxic control of the development of the surfactant system in the chicken: evidence for physiological heterokairy. AB - The surfactant system, a complex mixture of lipids and proteins, controls surface tension in the lung and is crucial for the first breath at birth, and thereafter. Heterokairy is defined as plasticity of a developmental process within an individual. Here, we provide experimental evidence for the concept of heterokairy, as hypoxia induces a change in the onset and rate of development of surfactant, probably via endogenous glucocorticoids, to produce individuals capable of surviving early hatching. Chicken eggs were incubated under normoxic (21% O(2)) conditions throughout or under hypoxic (17% O(2)) conditions from day 10 of incubation. Embryos were sampled at days 16, 18, and 20 and also 24 h after hatching. In a second experiment, dexamethasone (Dex), tri-iodothyronine (T(3)), or a combination (Dex + T(3)) was administered 24 and 48 h before each time point. Both hypoxia and Dex accelerated maturation of the surfactant lipids by increasing total phospholipid (PL), disaturated phospholipid (DSP), and cholesterol (Chol) in lavage at days 16 and 18. Maturation of surfactant lipid composition was accelerated, with day 16 %DSP/PL, Chol/DSP, and Chol/PL resembling the ratios of day 20 control animals. The effect of Dex + T(3) was similar to that of Dex alone. Hypoxia increased plasma corticosterone levels at day 16, while plasma T(3) levels were not affected. Hence, exposure to hypoxia during critical developmental windows accelerates surfactant maturation, probably by increasing corticosterone production. This internal modulation of the developmental response to an external stimulus is a demonstration of physiological heterokairy. PMID- 15117726 TI - Spontaneous beat-by-beat fluctuations of total peripheral and cerebrovascular resistance in response to tilt. AB - Beat-by-beat estimates of total peripheral resistance (TPR) can be obtained from continuous measurements of cardiac output by using Doppler ultrasound and noninvasive mean arterial blood pressure (MAP). We employed transfer function analysis to study the heart rate (HR) and vascular response to spontaneous changes in blood pressure from the relationships of systolic blood pressure (SBP) to HR (SBP-->HR), MAP to total peripheral resistance (TPR) and cerebrovascular resistance index (CVRi) (MAP-->TPR and MAP-->CVRi), as well as stroke volume (SV) to TPR in nine healthy subjects in supine and 45 degrees head-up tilt positions. The gain of the SBP-->HR transfer function was reduced with tilt in both the low- (0.03-0.15 Hz) and high-frequency (0.15-0.35 Hz) regions. In contrast, MAP-->TPR transfer function gain was not affected by head-up tilt, but it did increase from low- to high-frequency regions. The phase relationships between MAP-->TPR were unaffected by head-up tilt, but, consistent with an autoregulatory system, changes in MAP were followed by directionally similar changes in TPR, just as observed for the MAP-->CVRi. The SV-->TPR had high coherence with a constant phase of 150-160 degrees. Together, these data that showed changes in MAP preceded changes in TPR, as well as a possible link between SV and TPR, are consistent with complex interactions between the vascular component of the arterial and cardiopulmonary baroreflexes and intrinsic properties such as the myogenic response of the resistance arteries. PMID- 15117727 TI - Comparison of effects of exercise and diuretic on left ventricular geometry, mass, and insulin resistance in older hypertensive adults. AB - To compare the effects of exercise training and hydrochlorothiazide on left ventricular (LV) geometry and mass, blood pressure (BP), and hyperinsulinemia in older hypertensive adults, we studied 28 patients randomized either to a group (age 66.4 +/- 1.3 yr; n = 16) that exercised or to a group (age 65.3 +/- 1.2 yr; n = 12) that received hydrochlorothiazide for 6 mo. Endurance exercise training induced a 15% increase in peak aerobic power. The reduction in systolic BP was twofold greater with thiazide than with exercise (26.6 +/- 12.2 vs. 11.5 +/- 10.9 mmHg). Exercise and thiazide reduced LV wall thickness, LV mass index (14% in each group), and the LV wall thickness-to-radius ratio (h/r) similarly (exercise: before 0.48 +/- 0.2, after 0.42 +/- 0.01; thiazide: before 0.47 +/- 0.04, after 0.40 +/- 0.04; P = 0.017). The reductions in systolic BP and h/r were correlated in the exercise group (r = 0.70, P = 0.005) but not in the thiazide group. Exercise training reduced glucose-stimulated hyperinsulinemia (before: 13.65 +/- 2.6 vs. 9.84 +/- 1.5 mU.ml(-1).min; P = 0.04) and insulin resistance. Thiazide did not affect plasma insulin levels. The results suggest that although exercise is less effective in reducing systolic BP than thiazide, it can induce regression of LV hypertrophy similar in magnitude to thiazide. Unlike hydrochlorothiazide, exercise training can improve insulin resistance and aerobic capacity in older hypertensive people. PMID- 15117728 TI - Effect of intracerebroventricular angiotensin II on body weight and food intake in adult rats. AB - We recently reported that intracerebroventricular infusions of ANG II decreased food intake and increased energy expenditure in young rats. The aim of the present study was to determine if intracerebroventricular ANG II has similar effects in adult rats. The time course of the effect was also investigated with the idea that at earlier time points, a potential role for increased hypothalamic expression of corticotropin-releasing hormone (CRH) in the anorexia could be established. Finally, the contribution of ANG II-induced water drinking to the decrease in food intake was directly investigated. Rats received intracerebroventricular saline or ANG II using osmotic minipumps. Food intake, water intake, and body weight were measured daily. Experiments were terminated 2, 5, or 11 days after the beginning of the infusions. ANG II (approximately 32 ng.kg(-1).min(-1)) produced a transient decrease in food intake that lasted for 4 5 days although body weight continued to be decreased for the entire experiment most likely due to increased energy expenditure as evidenced by increased uncoupling protein-1 mRNA expression in brown adipose tissue. At 11 and 5 days, the expression of CRH mRNA was decreased. At 2 days, CRH expression was not suppressed even though body weight was decreased. The decrease in food intake and body weight was identical whether or not rats were allowed to increase water consumption. These data suggest that in adult rats ANG II acts within the brain to affect food intake and energy expenditure in a manner that is not related to water intake. PMID- 15117729 TI - Novel method for measuring effects of gas compression on expiratory flow. AB - During forced vital capacity maneuvers in subjects with expiratory flow limitation, lung volume decreases during expiration both by air flowing out of the lung (i.e., exhaled volume) and by compression of gas within the thorax. As a result, a flow-volume loop generated by using exhaled volume is not representative of the actual flow-volume relationship. We present a novel method to take into account the effects of gas compression on flow and volume in the first second of a forced expiratory maneuver (FEV(1)). In addition to oral and esophageal pressures, we measured flow and volume simultaneously using a volume displacement plethysmograph and a pneumotachograph in normal subjects and patients with expiratory flow limitation. Expiratory flow vs. plethysmograph volume signals was used to generate a flow-volume loop. Specialized software was developed to estimate FEV(1) corrected for gas compression (NFEV(1)). We measured reproducibility of NFEV(1) in repeated maneuvers within the same session and over a 6-mo interval in patients with chronic obstructive pulmonary disease. Our results demonstrate that NFEV(1) significantly correlated with FEV(1), peak expiratory flow, lung expiratory resistance, and total lung capacity. During intrasession, maneuvers with the highest and lowest FEV(1) showed significant statistical difference in mean FEV(1) (P < 0.005), whereas NFEV(1) from the same maneuvers were not significantly different from each other (P > 0.05). Furthermore, variability of NFEV(1) measurements over 6 mo was <5%. We concluded that our method reliably measures the effect of gas compression on expiratory flow. PMID- 15117730 TI - Chronic in ovo hypoxia decreases pulmonary arterial contractile reactivity and induces biventricular cardiac enlargement in the chicken embryo. AB - Although chronic prenatal hypoxia is considered a major cause of persistent pulmonary hypertension of the newborn, experimental studies have failed to consistently find pulmonary hypertensive changes after chronic intrauterine hypoxia. We hypothesized that chronic prenatal hypoxia induces changes in the pulmonary vasculature of the chicken embryo. We analyzed pulmonary arterial reactivity and structure and heart morphology of chicken embryos maintained from days 6 to 19 of the 21-day incubation period under normoxic (21% O(2)) or hypoxic (15% O(2)) conditions. Hypoxia increased mortality (0.46 vs. 0.14; P < 0.01) and reduced the body mass of the surviving 19-day embryos (22.4 +/- 0.5 vs. 26.6 +/- 0.7 g; P < 0.01). A decrease in the response of the pulmonary artery to KCl was observed in the 19-day hypoxic embryos. The contractile responses to endothelin 1, the thromboxane A(2) mimetic U-46619, norepinephrine, and electrical-field stimulation were also reduced in a proportion similar to that observed for KCl induced contractions. In contrast, no hypoxia-induced decrease of response to vasoconstrictors was observed in externally pipped 21-day embryos (incubated under normoxia for the last 2 days). Relaxations induced by ACh, sodium nitroprusside, or forskolin were unaffected by chronic hypoxia in the pulmonary artery, but femoral artery segments of 19-day hypoxic embryos were significantly less sensitive to ACh than arteries of control embryos [pD(2) (= -log EC(50)): 6.51 +/- 0.1 vs. 7.05 +/- 0.1, P < 0.01]. Pulmonary vessel density, percent wall area, and periarterial sympathetic nerve density were not different between control and hypoxic embryos. In contrast, hypoxic hearts showed an increase in right and left ventricular wall area and thickness. We conclude that, in the chicken embryo, chronic moderate hypoxia during incubation transiently reduced pulmonary arterial contractile reactivity, impaired endothelium-dependent relaxation of femoral but not pulmonary arteries, and induced biventricular cardiac hypertrophy. PMID- 15117731 TI - Apolipoprotein A-IV stimulates duodenal vagal afferent activity to inhibit gastric motility via a CCK1 pathway. AB - Apolipoprotein A-IV (apo A-IV), a peptide expressed by enterocytes in the mammalian small intestine and released in response to long-chain triglyceride absorption, may be involved in the regulation of gastric acid secretion and gastric motility. The specific aim of the present study was to determine the pathway involved in mediating inhibition of gastric motility produced by apo A IV. Gastric motility was measured manometrically in response to injections of either recombinant purified apo A-IV (200 microg) or apo A-I, the structurally similar intestinal apolipoprotein not regulated by triglyceride absorption, close to the upper gastrointestinal tract in urethane-anesthetized rats. Injection of apo A-IV significantly inhibited gastric motility compared with apo A-I or vehicle injections. The response to exogenous apo A-IV injections was significantly reduced by 77 and 55%, respectively, in rats treated with the CCK(1) receptor blocker devazepide or after functional vagal deafferentation by perineural capsaicin treatment. In electrophysiological experiments, isolated proximal duodenal vagal afferent fibers were recorded in vitro in response to close-arterial injection of vehicle, apo A-IV (200 microg), or CCK (10 pmol). Apo A-IV stimulated the discharge of duodenal vagal afferent fibers, significantly increasing the discharge in 4/7 CCK-responsive units, and the response was abolished by CCK(1) receptor blockade with devazepide. These data suggest that apo A-IV released from the intestinal mucosa during lipid absorption stimulates the release of endogenous CCK that activates CCK(1) receptors on vagal afferent nerve terminals initiating feedback inhibition of gastric motility. PMID- 15117732 TI - AIF-1 expression modulates proliferation of human vascular smooth muscle cells by autocrine expression of G-CSF. AB - OBJECTIVE: Allograft inflammatory factor-1 (AIF-1) is associated with vascular smooth muscle cell (VSMC) activation and vascular injury. The purpose of this study was to characterize the molecular mechanism of AIF-1 growth-enhancing effects in human VSMC. METHODS AND RESULTS: Primary human VSMCs were stably transduced with AIF-1 retrovirus (RV). Impact on cell growth was evaluated by the increase in cell number, and the effects on gene expression were determined by cDNA microarray analysis. AIF-RV overexpressing cells grew significantly more rapidly than empty-RV control cells in growth medium and serum-reduced medium (P<0.01 and 0.02, respectively). cDNA microarray analysis and Western blotting on serum-starved AIF-1-transduced VSMCs identified increased mRNA expression of several cell cycle proteins and, surprisingly, the cytokine G-CSF. Addition of G CSF caused a 75% increase in proliferation of VSMCs in the absence of serum growth factors. The proliferative effects of AIF-1 were abrogated by neutralizing antibodies to G-CSF (P<0.05), and AIF-1-transduced VSMCs are chemotactic for human monocytes. Increased expression of G-CSF and colocalization with AIF-1 positive cells were seen in diseased, not normal human coronary arteries. CONCLUSIONS: This study indicates that AIF-1 enhances VSMC growth by autocrine production of G-CSF, and AIF-1 expression may influence VSMC-inflammatory cell communication. PMID- 15117733 TI - Unique morphology and focal adhesion development of valvular endothelial cells in static and fluid flow environments. AB - BACKGROUND: The influence of mechanical forces on cell function has been well documented for many different cell types. Endothelial cells native to the aortic valve may play an important role in mediating tissue responses to the complex fluid environment, and may therefore respond to fluid flow in a different manner than more characterized vascular endothelial cells. METHODS AND RESULTS: Porcine endothelial cells of aortic and aortic valvular origin were subjected to 20 dynes/cm2 steady laminar shear stress for up to 48 hours, with static cultures serving as controls. The aortic valve endothelial cells were observed to align perpendicular to flow, in direct contrast to the aortic endothelial cells, which aligned parallel to flow. Focal adhesion complexes reorganized prominently at the ends of the long axis of aligned cells. Valvular endothelial cell alignment was dependent on Rho-kinase signaling, whereas vascular endothelial cell alignment was dependent on both Rho-kinase and phosphatidylinositol 3-kinase signal pathways. CONCLUSIONS: These differences in response to mechanical forces suggest a unique phenotype of valvular endothelial cells not mimicked by vascular endothelial cells, and could have implications for cardiovascular cell biology and cell-source considerations for tissue-engineered valvular substitutes. PMID- 15117734 TI - Analysis of apolipoprotein A5, c3, and plasma triglyceride concentrations in genetically engineered mice. AB - OBJECTIVE: Both the apolipoprotein A5 and C3 genes have repeatedly been shown to play an important role in determining plasma triglyceride concentrations in humans and mice. In mice, transgenic and knockout experiments indicate that plasma triglyceride levels are strongly altered by changes in the expression of either of these 2 genes. In humans, common polymorphisms in both genes have also been associated with plasma triglyceride concentrations. These similar findings raised the issue of the relationship between these 2 genes and altered triglycerides. METHODS AND RESULTS: To address this issue, we generated independent lines of mice that either overexpressed ("double transgenic") or completely lacked ("double knockout") both apolipoprotein genes. We report that both "double transgenic" and "double knockout" mice display normal triglyceride concentrations compared with overexpression or deletion of either gene alone. Furthermore, we find that human ApoAV plasma protein levels in the "double transgenic" mice are approximately 500-fold lower than human ApoCIII levels, supporting ApoAV as a potent triglyceride modulator despite its low concentration. CONCLUSIONS: Together, these data support that APOA5 and APOC3 independently influence plasma triglyceride concentrations but in an opposing manner. PMID- 15117735 TI - R-cadherin:beta-catenin complex and its association with vascular smooth muscle cell proliferation. AB - OBJECTIVE: Vascular smooth muscle cell (VSMC) proliferation is an important component of atherosclerosis, restenosis after angioplasty and stent placement, and vein graft failure. Outside-in signaling from the cadherin:beta-catenin complex can increase transcription of the cell-cycle gene cyclin D1; however, its role in VSMC proliferation has only recently been considered. METHODS AND RESULTS: We examined the involvement of R-cadherin and beta-catenin in VSMC proliferation in balloon-injured carotid arteries in vivo and aortic rings in vitro. The number of medial VSMCs positive for R-cadherin was significantly reduced by 32%+/-5%, 52%+/-10%, and 23%+/-2% at 0.25, 24, and 48 hours after injury in vivo, respectively. These changes in cadherin expression coincided with the detection of nuclear beta-catenin and elevated cyclin D1 expression. Furthermore, loss of R-cadherin expression was associated with medial VSMC proliferation. Inhibition of classical cadherin function with a HAV peptide and R cadherin neutralizing antibodies significantly increased proliferation by 4.3+/ 1.0-fold and 4.1+/-0.98-fold, and increased the number of cells with beta-catenin in the nucleus and expressing cyclin D1 in aortic rings. CONCLUSIONS: These results suggest that R-cadherin expression and beta-catenin signaling may be associated with increased cyclin D1 expression and VSMC proliferation and may therefore play an important role in vascular disease. PMID- 15117736 TI - Role of tissue factor in hemostasis, thrombosis, and vascular development. AB - Tissue factor (TF) is best known as the primary cellular initiator of blood coagulation. After vessel injury, the TF:FVIIa complex activates the coagulation protease cascade, which leads to fibrin deposition and activation of platelets. TF deficiency causes embryonic lethality in the mouse and there have been no reports of TF deficiency in humans. These results indicate that TF is essential for life, most likely because of its central role in hemostasis. In addition, aberrant TF expression within the vasculature initiates life-threatening thrombosis in various diseases, such as sepsis, atherosclerosis, and cancer. Finally, recent studies have revealed a nonhemostatic role of TF in the generation of coagulation proteases and subsequent activation of protease activated receptors (PARs) on vascular cells. This TF-dependent signaling contributes to a variety of biological processes, including inflammation, angiogenesis, metastasis, and cell migration. This review focuses on the roles of TF in hemostasis, thrombosis, and vascular development. PMID- 15117737 TI - Susceptibility to early atherosclerosis in male mice is mediated by estrogen receptor alpha. AB - OBJECTIVE: Vascular tissues express 2 types of estrogen receptors (ERs): ERalpha and ERbeta. Their role in early atherosclerosis remains poorly understood, particularly in males. We developed and characterized an atherosclerosis model in ERalpha knockout male mice to investigate directly its role in atheroma. METHODS AND RESULTS: Cholesterol-fed ERalpha knockout and wild-type mice developed early atheroma characterized by fatty streaks and foam cells. ERalpha wild-type mice developed 3.8-fold greater lesion area, more advanced lesions, more extensive lesion distribution, twice the number of lesions, and at a 2.2-fold faster rate than ERalpha knockout mice. Lesion development and atheroma susceptibility in ERalpha wild-type and knockout mice were independent of serum cholesterol, triglycerides, high-density lipoproteins, 17beta-estradiol, and testosterone levels. In contrast, castration eliminated the predilection of ERalpha wild-type mice for atheroma, suggesting that testosterone mediates ERalpha-dependent atheroma formation in males. CONCLUSIONS: This study is the first to report that the ERalpha mediates susceptibility to early atherosclerosis in male mice by a testosterone-dependent pathway, suggesting that local production of estrogen from testosterone in the vessel wall may promote atheroma formation in ERalpha males. Our findings may have implications for selective targeting of ERalpha in atherosclerotic disease. PMID- 15117738 TI - Hypoxia-induced mitogenic factor has antiapoptotic action and is upregulated in the developing lung: coexpression with hypoxia-inducible factor-2alpha. AB - Hypoxia-induced mitogenic factor (HIMF), also called FIZZ1 or RELMalpha, was a newly found cytokine. Hypoxia caused robust HIMF induction in the lung, and HIMF has potent pulmonary vasoconstrictive, proliferative, and angiogenic properties. To investigate the role of HIMF in lung development, we determined its spatial and temporal expression. From embryonic day (E)16 to postnatal day (P)28, HIMF was strongly expressed in the cytoplasm of bronchial epithelial cells, type II cells, endothelial cells, and primitive mesenchymal cells. Treatment with HIMF resulted in a significant reduction of apoptosis in cultured embryonic lung, thus revealing a previously unknown function of HIMF. Because HIMF gene is upregulated by hypoxia and contains a hypoxia-inducible transcription factor (HIF) binding site, we subsequently investigated whether HIMF was coexpressed with HIF-2alpha or HIF-1alpha. HIF-1alpha expression was temporally distinct from HIMF expression. In contrast, HIF-2alpha was present in endothelial cells, bronchial epithelial cells, and type II cells from E18 to P28. Thus, HIMF and HIF-2alpha were temporally and spatially coexpressed in the developing lung. These results indicate a role for HIMF in lung development, possibly under the control of HIF 2, and suggest that HIMF regulates apoptosis and may participate in lung alveolarization and maturation. PMID- 15117739 TI - Angiotensin converting enzyme genotype and strength in chronic obstructive pulmonary disease. AB - Quadriceps muscle weakness is an important contributor to exercise limitation in patients with chronic obstructive pulmonary disease. The deletion allele of the angiotensin converting enzyme gene polymorphism has previously been associated with a greater response to strength training in healthy subjects and might, therefore, protect against detraining in these patients. In 103 stable outpatients (mean [SD] FEV(1) 34.4 [16.5] % predicted), the angiotensin deletion allele was associated with greater isometric quadriceps strength; mean (SD) 31.4 (10.8) kg for insertion homozygotes, 34.1 (13.0) kg for heterozygotes, and 38.3 (11.6) kg for deletion homozygotes (p = 0.04 linear trend). Adjusted for fat-free mass, the relationship was stronger (linear trend p = 0.007). There was no correlation between strength and genotype in a group of 101 age-matched healthy control subjects. Twitch quadriceps force in response to magnetic femoral nerve stimulation, measured in 39 patients, was also genotype dependent; 8.3 (2.6) kg for insertion homozygotes, 10.1 (3.6) kg for heterozygotes, and 12.4 (3.5) kg for deletion homozygotes (p = 0.002 linear trend). Body mass index and fat-free mass did not differ significantly between genotypes in either group. There was no association in either patients or control subjects between genotype and inspiratory muscle strength. In chronic obstructive pulmonary disease the deletion allele is associated with greater quadriceps strength independent of confounding factors. PMID- 15117740 TI - Dietary fiber and reduced cough with phlegm: a cohort study in Singapore. AB - Smoking is the major risk factor for chronic respiratory symptoms, but dietary factors may also play a role. Most studies of diet and lung disease have been cross-sectional and conducted in populations with a Western-style diet. We analyzed the relation between dietary intake at baseline and new onset of cough with phlegm in a population-based cohort of 63,257 middle-aged Chinese men and women initiated in Singapore between 1993 and 1998. Beginning in 1999, we ascertained respiratory symptoms by telephone interview and have identified 571 incident cases of cough with phlegm among the 49,140 cohort members with completed follow-up. Nonstarch polysaccharides, a major component of dietary fiber, total fruit, and soy isoflavones had the strongest associations. Odds ratios comparing highest and lowest quartiles after adjustment for age, sex, dialect group, total energy intake, and smoking were 0.61 (95% confidence interval [CI]: 0.47, 0.78; p for trend < 0.001) for nonstarch polysaccharides, 0.67 (95% CI: 0.52, 0.87; p for trend = 0.006) for fruit, and 0.67 (95% CI: 0.53, 0.86; p for trend = 0.001) for soy isoflavones. These data suggest that a diet high in fiber from fruit and, possibly, soyfoods may reduce the incidence of chronic respiratory symptoms. Associated nutrients, such as flavonoids, may contribute to this association. PMID- 15117741 TI - Association of chronic obstructive pulmonary disease severity and Pneumocystis colonization. AB - Factors modulating the variable progression of chronic obstructive pulmonary disease (COPD) are largely unknown, but infectious agents may play a role. Because Pneumocystis has previously been shown to induce a CD8(+) lymphocyte- and neutrophil-predominant response similar to that in COPD, we explored the association of the organism with accelerated disease progression. We examined Pneumocystis colonization rates in lung tissue obtained during lung resection or transplantation in smokers with a range of airway obstruction severity and in a control group with lung diseases other than COPD. Using nested polymerase chain reaction, Pneumocystis colonization was detected in 36.7% of patients with very severe COPD (Global Health Initiative on Obstructive Lung Disease [GOLD] Stage IV) compared with 5.3% of smokers with normal lung function or less severe COPD (Stages 0, I, II, and III) (p = 0.004) and with 9.1% of control subjects (p = 0.007). Colonized subjects exhibited more severe airway obstruction (median FEV(1) = 21% predicted versus 62% in noncolonized subjects, p = 0.006). GOLD IV was the strongest predictor of Pneumocystis colonization (odds ratio = 7.3, 95% confidence interval = 2.4-22.4, p < 0.001) and was independent of smoking history. We conclude that there is a strong association between Pneumocystis colonization and severity of airflow obstruction in smokers, suggesting a possible pathogenic link with COPD progression. PMID- 15117742 TI - Persistent colonization by Haemophilus influenzae in chronic obstructive pulmonary disease. AB - Nontypeable Haemophilus influenzae colonizes the respiratory tract of adults with chronic obstructive pulmonary disease (COPD) and causes intermittent exacerbations. Isolates of H. influenzae collected monthly in a prospective study were subjected to molecular typing. During a 7-year study spanning 345 patient months of observation, 122 episodes of negative cultures lasting 1 month or more, and that were preceded and followed by isolation of an apparently identical strain of H. influenzae, were found. Seventeen such episodes of negative cultures, lasting 6 months or more and spanning 203 patient-months, were studied in detail to test the hypothesis that these periods of negative cultures represented continuous colonization by the same strain of H. influenzae. Molecular typing by three independent methods established that the strains preceding and following the episodes of negative cultures were indeed identical. Strain-specific H. influenzae DNA was detected in some of the sputum samples that had yielded negative cultures. These results indicate that some patients with COPD are persistently colonized with H. influenzae and that sputum cultures underestimate the frequency of colonization of the respiratory tract by H. influenzae in COPD. This observation has a significant impact on understanding bacterial colonization in COPD. PMID- 15117743 TI - Differential cytokine gene expression in the diaphragm in response to strenuous resistive breathing. AB - Strenuous resistive breathing induces plasma cytokines that do not originate from circulating monocytes. We hypothesized that cytokine production is induced inside the diaphragm in response to resistive loading. Anesthetized, tracheostomized, spontaneously breathing Sprague-Dawley rats were subjected to 1, 3, or 6 hours of inspiratory resistive loading, corresponding to 45-50% of the maximum inspiratory pressure. Unloaded sham-operated rats breathing spontaneously served as control animals. The diaphragm and the gastrocnemius muscles were excised at the end of the loading period, and messenger ribonucleic acid expression of tumor necrosis factor-alpha, tumor necrosis factor-beta, interleukin (IL)-1alpha, IL-1beta, IL 2, IL-3, IL-4, IL-5, IL-6, IL-10, IFN-gamma, and two housekeeping genes was analyzed using multiprobe RNase protection assay. IL-6, IL-1beta, and, to lesser extents, tumor necrosis factor-alpha, IL-10, IFN-gamma, and IL-4 were significantly increased in a time-dependent fashion in the diaphragms but not the gastrocnemius of loaded animals or in the diaphragm of control animals. Elevation of protein levels of IL-6 and IL-1beta in the diaphragm of loaded animals was confirmed with immunoblotting. Immunostaining revealed IL-6 protein localization inside diaphragmatic muscle fibers. We conclude that increased ventilatory muscle activity during resistive loading induces differential elevation of proinflammatory and antiinflammatory cytokine gene expression in the ventilatory muscles. PMID- 15117744 TI - Pigment epithelium-derived factor in idiopathic pulmonary fibrosis: a role in aberrant angiogenesis. AB - Pigment epithelium-derived factor (PEDF) is a 50-kD protein with angiostatic and neurotrophic activities that regulates vascular development within the eye. PEDF expression was increased in the lungs of patients with idiopathic pulmonary fibrosis (IPF) based on microarray analyses. Angiogenesis has been implicated in the pathogenesis of fibrotic lung diseases, we therefore hypothesized that regional abnormalities in vascularization occur in IPF as a result of an imbalance between PEDF and vascular endothelial growth factor. We demonstrated that vascular density is regionally decreased in IPF within the fibroblastic foci, and that within these areas PEDF was increased, whereas vascular endothelial growth factor was decreased. PEDF colocalized with the fibrogenic cytokine, transforming growth factor (TGF)-beta 1, particularly within the fibrotic interstitium and the fibroblastic focus, and prominently within the epithelium directly overlying the fibroblastic focus. This suggested that TGF beta 1 might regulate PEDF expression. Using 3T3-L1 fibroblasts and human lung fibroblasts, we showed that PEDF was indeed a TGF-beta 1 target gene. Collectively, our findings implicate PEDF as a regulator of pulmonary angiogenesis and an important mediator in IPF. PMID- 15117745 TI - Discordant extracellular superoxide dismutase expression and activity in neonatal hyperoxic lung. AB - Antioxidant defenses in the neonatal lung are required to adapt to the oxygen (O(2))-rich postnatal environment, and oxidant/antioxidant imbalance is a predisposition to lung injury when high concentrations of inspired O(2) are used in neonatal lung diseases. The lung's main extracellular enzymatic defense against superoxide, extracellular superoxide dismutase (EC-SOD), is closely regulated during development. In testing the hypothesis that developmental change in EC-SOD expression and activity in the immature lung would be disrupted by hyperoxia, we found a doubling of lung EC-SOD protein in newborn rats exposed to 95% O(2) for 1 week. Furthermore, EC-SOD protein secretion increased, but EC-SOD enzyme activity did not change with O(2) exposure. EC-SOD mRNA did not change at multiple points between 6 hours and 8 days. Lung EC-SOD recovered by immunoprecipitation after 1 week of O(2) showed strong increases in protein nitrotyrosine and variable, nonsignificant differences in protein carbonyl content. These data provide the first direct evidence that EC-SOD is itself a target of nitration in hyperoxia, and offer a plausible explanation for low EC SOD activity despite its increased secretion by O(2)-exposed neonatal lung. PMID- 15117746 TI - A prospective study of Fel d1 and Der p1 exposure in infancy and childhood wheezing. AB - The impact of domestic exposure to cat allergen (Fel d1) and house dust mite (Der p1) on wheezing from birth to the age of 4 years was investigated in a multicenter prospective birth cohort; 1,611 mothers were recruited before delivery in Ashford, England, and Barcelona and Menorca, Spain. Exposures were gathered via dust sample collection at children's home in their first year of life. Families provided complete outcome data (wheezing status in all 4 years) for 1,289 children. Domestic allergen levels varied substantially between centers. Six hundred three (47%) children never wheezed during their first 4 years of life. Der p1 did not correlate with any type of wheezing outcome. Fel d1 significantly increased the risk of wheezing in 3- and 4-year-olds in comparison to 1-year-olds. Distinct risk profiles were found for wheezing at different ages. Multivariate analysis revealed an interaction between Fel d1 and maternal asthma among children who wheeze in Year 4 (relative risk = 2.77; 95% confidence interval = 1.19-6.46). Our data support the idea that several patterns of wheezing with different risk profiles exist among young children. The effect of Fel d1 exposure varied according to age and maternal asthma. PMID- 15117747 TI - A link between lung androgen metabolism and the emergence of mature epithelial type II cells. AB - Lung maturation is delayed in male fetuses compared with female fetuses, which has been attributed to higher levels of androgens in the male lung. Our previous studies demonstrated that the genes encoding for the 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) type 5 (androstenedione --> testosterone) and type 2 (the opposite reaction) are expressed in human epithelial type II (PTII)-like A549 cells and in human lung fibroblasts, respectively. Here, we aim to explain the physiological relevance of androgen synthesis by PTII cells. We showed that both 17 beta-HSD type 2 and type 5 genes are upregulated in correlation with the emergence of mature PTII cells in both male and female developing lungs of the mouse. In contrast, the androgen receptor gene is expressed equally in both sexes with no temporal regulation. We conclude that the expression profile of the 17 beta-HSD type 5 gene does not explain the presence of higher levels of androgen in the male fetal lung, but that androgen synthesis must be a normal feature of mature PTII cells for both sexes. The production of androgens after the emergence of mature PTII cells should negatively regulate PTII cell maturation and, thus, a role for androgens in cell reprogramming is suggested. PMID- 15117748 TI - YAdumper: extracting and translating large information volumes from relational databases to structured flat files. AB - Downloading the information stored in relational databases into XML and other flat formats is a common task in bioinformatics. This periodical dumping of information requires considerable CPU time, disk and memory resources. YAdumper has been developed as a purpose-specific tool to deal with the integral structured information download of relational databases. YAdumper is a Java application that organizes database extraction following an XML template based on an external Document Type Declaration. Compared with other non-native alternatives, YAdumper substantially reduces memory requirements and considerably improves writing performance. PMID- 15117749 TI - HPID: the Human Protein Interaction Database. AB - The Human Protein Interaction Database (http://www.hpid.org) was designed (1) to provide human protein interaction information pre-computed from existing structural and experimental data, (2) to predict potential interactions between proteins submitted by users and (3) to provide a depository for new human protein interaction data from users. Two types of interaction are available from the pre computed data: (1) interactions at the protein superfamily level and (2) those transferred from the interactions of yeast proteins. Interactions at the superfamily level were obtained by locating known structural interactions of the PDB in the SCOP domains and identifying homologs of the domains in the human proteins. Interactions transferred from yeast proteins were obtained by identifying homologs of the yeast proteins in the human proteins. For each human protein in the database and each query submitted by users, the protein superfamilies and yeast proteins assigned to the protein are shown, along with their interacting partners. We have also developed a set of web-based programs so that users can visualize and analyze protein interaction networks in order to explore the networks further. AVAILABILITY: http://www.hpid.org. PMID- 15117750 TI - SIMCOAL 2.0: a program to simulate genomic diversity over large recombining regions in a subdivided population with a complex history. AB - We present an extension of the program SIMCOAL, which allows for simulation of the genomic diversity of samples drawn from a set of populations with arbitrary patterns of migrations and complex demographic histories, including bottlenecks and various modes of demographic expansion. The main additions to the previous version include the possibility of arbitrary and heterogeneous recombination rates between adjacent loci and multiple coalescent events per generation, allowing for the simulation of very large samples and recombining genomic regions, together with the simulation of single nucleotide polymorphism data with frequency ascertainment bias. AVAILABILITY: http://cmpg.unibe.ch/software/simcoal2/. PMID- 15117751 TI - Efficient combination of multiple word models for improved sequence comparison. AB - MOTIVATION: Studies of efficient and sensitive sequence comparison methods are driven by a need to find homologous regions of weak similarity between large genomes. RESULTS: We describe an improved method for finding similar regions between two sets of DNA sequences. The new method generalizes existing methods by locating word matches between sequences under two or more word models and extending word matches into high-scoring segment pairs (HSPs). The method is implemented as a computer program named DDS2. Experimental results show that DDS2 can find more HSPs by using several word models than by using one word model. AVAILABILITY: The DDS2 program is freely available for academic use in binary code form at http://bioinformatics.iastate.edu/aat/align/align.html and in source code form from the corresponding author. PMID- 15117752 TI - Interactively optimizing signal-to-noise ratios in expression profiling: project specific algorithm selection and detection p-value weighting in Affymetrix microarrays. AB - MOTIVATION: The most commonly utilized microarrays for mRNA profiling (Affymetrix) include 'probe sets' of a series of perfect match and mismatch probes (typically 22 oligonucleotides per probe set). There are an increasing number of reported 'probe set algorithms' that differ in their interpretation of a probe set to derive a single normalized 'signal' representative of expression of each mRNA. These algorithms are known to differ in accuracy and sensitivity, and optimization has been done using a small set of standardized control microarray data. We hypothesized that different mRNA profiling projects have varying sources and degrees of confounding noise, and that these should alter the choice of a specific probe set algorithm. Also, we hypothesized that use of the Microarray Suite (MAS) 5.0 probe set detection p-value as a weighting function would improve the performance of all probe set algorithms. RESULTS: We built an interactive visual analysis software tool (HCE2W) to test and define parameters in Affymetrix analyses that optimize the ratio of signal (desired biological variable) versus noise (confounding uncontrolled variables). Five probe set algorithms were studied with and without statistical weighting of probe sets using the MAS 5.0 probe set detection p-values. The signal-to-noise ratio optimization method was tested in two large novel microarray datasets with different levels of confounding noise, a 105 sample U133A human muscle biopsy dataset (11 groups: mutation-defined, extensive noise), and a 40 sample U74A inbred mouse lung dataset (8 groups: little noise). Performance was measured by the ability of the specific probe set algorithm, with and without detection p value weighting, to cluster samples into the appropriate biological groups (unsupervised agglomerative clustering with F-measure values). Of the total random sampling analyses, 50% showed a highly statistically significant difference between probe set algorithms by ANOVA [F(4,10) > 14, p < 0.0001], with weighting by MAS 5.0 detection p-value showing significance in the mouse data by ANOVA [F(1,10) > 9, p < 0.013] and paired t-test [t(9) = -3.675, p = 0.005]. Probe set detection p-value weighting had the greatest positive effect on performance of dChip difference model, ProbeProfiler and RMA algorithms. Importantly, probe set algorithms did indeed perform differently depending on the specific project, most probably due to the degree of confounding noise. Our data indicate that significantly improved data analysis of mRNA profile projects can be achieved by optimizing the choice of probe set algorithm with the noise levels intrinsic to a project, with dChip difference model with MAS 5.0 detection p value continuous weighting showing the best overall performance in both projects. Furthermore, both existing and newly developed probe set algorithms should incorporate a detection p-value weighting to improve performance. AVAILABILITY: The Hierarchical Clustering Explorer 2.0 is available at http://www.cs.umd.edu/hcil/hce/ Murine arrays (40 samples) are publicly available at the PEPR resource (http://microarray.cnmcresearch.org/pgadatatable.asp http://pepr.cnmcresearch.org Chen et al., 2004). PMID- 15117753 TI - Class discovery and classification of tumor samples using mixture modeling of gene expression data--a unified approach. AB - MOTIVATION: The DNA microarray technology has been increasingly used in cancer research. In the literature, discovery of putative classes and classification to known classes based on gene expression data have been largely treated as separate problems. This paper offers a unified approach to class discovery and classification, which we believe is more appropriate, and has greater applicability, in practical situations. RESULTS: We model the gene expression profile of a tumor sample as from a finite mixture distribution, with each component characterizing the gene expression levels in a class. The proposed method was applied to a leukemia dataset, and good results are obtained. With appropriate choices of genes and preprocessing method, the number of leukemia types and subtypes is correctly inferred, and all the tumor samples are correctly classified into their respective type/subtype. Further evaluation of the method was carried out on other variants of the leukemia data and a colon dataset. PMID- 15117754 TI - Identification of DNA regulatory motifs using Bayesian variable selection. AB - MOTIVATION: Understanding the mechanisms that determine gene expression regulation is an important and challenging problem. A common approach consists of identifying DNA-binding sites from a collection of co-regulated genes and their nearby non-coding DNA sequences. Here, we consider a regression model that linearly relates gene expression levels to a sequence matching score of nucleotide patterns. We use Bayesian models and stochastic search techniques to select transcription factor binding site candidates, as an alternative to stepwise regression procedures used by other investigators. RESULTS: We demonstrate through simulated data the improved performance of the Bayesian variable selection method compared to the stepwise procedure. We then analyze and discuss the results from experiments involving well-studied pathways of Saccharomyces cerevisiae and Schizosaccharomyces pombe. We identify regulatory motifs known to be related to the experimental conditions considered. Some of our selected motifs are also in agreement with recent findings by other researchers. In addition, our results include novel motifs that constitute promising sets for further assessment. AVAILABILITY: The Matlab code for implementing the Bayesian variable selection method may be obtained from the corresponding author. PMID- 15117755 TI - The database of epoxide hydrolases and haloalkane dehalogenases: one structure, many functions. AB - The epoxide hydrolases and haloalkane dehalogenases database (EH/HD) integrates sequence and structure of a highly diverse protein family, including mainly the Asp-hydrolases of EHs and HDs but also proteins, such as Ser-hydrolases non-heme peroxidases, prolyl iminopetidases and 2-hydroxymuconic semialdehyde hydrolases. These proteins have a highly conserved structure, but display a remarkable diversity in sequence and function. A total of 305 protein entries were assigned to 14 homologous families, forming two superfamilies. Annotated multisequence alignments and phylogenetic trees are provided for each homologous family and superfamily. Experimentally derived structures of 19 proteins are superposed and consistently annotated. Sequence and structure of all 305 proteins were systematically analysed. Thus, deeper insight is gained into the role of a highly conserved sequence motifs and structural elements. AVAILABILITY: The EH/HD database is available at http://www.led.uni-stuttgart.de PMID- 15117756 TI - A mixture model-based strategy for selecting sets of genes in multiclass response microarray experiments. AB - MOTIVATION: Multiclass response (MCR) experiments are those in which there are more than two classes to be compared. In these experiments, though the null hypothesis is simple, there are typically many patterns of gene expression changes across the different classes that led to complex alternatives. In this paper, we propose a new strategy for selecting genes in MCR that is based on a flexible mixture model for the marginal distribution of a modified F-statistic. Using this model, false positive and negative discovery rates can be estimated and combined to produce a rule for selecting a subset of genes. Moreover, the method proposed allows calculation of these rates for any predefined subset of genes. RESULTS: We illustrate the performance our approach using simulated datasets and a real breast cancer microarray dataset. In this latter study, we investigate predefined subset of genes and point out interesting differences between three distinct biological pathways. AVAILABILITY: http://www.bgx.org.uk/software.html PMID- 15117757 TI - Predicting allergenic proteins using wavelet transform. AB - MOTIVATION: With many transgenic proteins introduced today, the ability to predict their potential allergenicity has become an important issue. Previous studies were based on either sequence similarity or the protein motifs identified from known allergen databases. The similarity-based approaches, although being able to produce high recalls, usually have low prediction precisions. Previous motif-based approaches have been shown to be able to improve the precisions on cross-validation experiments. In this study, a system that combines the advantages of similarity-based and motif-based prediction is described. RESULTS: The new prediction system uses a clustering algorithm that groups the known allergenic proteins into clusters. Proteins within each cluster are assumed to carry one or more common motifs. After a multiple sequence alignment, proteins in each cluster go through a wavelet analysis program whereby conserved motifs will be identified. A hidden Markov model (HMM) profile will then be prepared for each identified motif. The allergens that do not appear to carry detectable allergen motifs will be saved in a small database. The allergenicity of an unknown protein may be predicted by comparing it against the HMM profiles, and, if no matching profiles are found, against the small allergen database by BLASTP. Over 70% of recall and over 90% of precision were observed using cross-validation experiments. Using the entire Swiss-Prot as the query, we predicted about 2000 potential allergens. AVAILABILITY: The software is available upon request from the authors. PMID- 15117758 TI - RRE: a tool for the extraction of non-coding regions surrounding annotated genes from genomic datasets. AB - RRE allows the extraction of non-coding regions surrounding a coding sequence [i.e. gene upstream region, 5'-untranslated region (5'-UTR), introns, 3'-UTR, downstream region] from annotated genomic datasets available at NCBI. AVAILABILITY: RRE parser and web-based interface are accessible at http://www.bioinformatica.unito.it/bioinformatics/rre/rre.html PMID- 15117759 TI - Genome wide identification and classification of alternative splicing based on EST data. AB - MOTIVATION: Alternative splicing is currently seen to explain the vast disparity between the number of predicted genes in the human genome and the highly diverse proteome. The mapping of expressed sequences tag (EST) consensus sequences derived from the GeneNest database onto the genome provides an efficient way of predicting exon-intron boundaries, gene structure and alternative splicing events. However, the alternative splicing events are obscured by a large number of putatively artificial exon boundaries arising due to genomic contamination or alignment errors. The current work describes a methodology to associate quality values to the predicted exon-intron boundaries. High quality exon-intron boundaries are used to predict constitutive and alternative splicing ranked by confidence values, aiming to facilitate large-scale analysis of alternative splicing and splicing in general. RESULTS: Applying the current methodology, constitutive splicing is observed in 33,270 EST clusters, out of which 45% are alternatively spliced. The classification derived from the computed confidence values for 17 of these splice events frequently correlate (15/17) with RT-PCR experiments performed for 40 different tissue samples. As an application of the confidence measure, an evaluation of distribution of alternative splicing revealed that majority of variants correspond to the coding regions of the genes. However, still a significant fraction maps to non-coding regions, thereby indicating a functional relevance of alternative splicing in untranslated regions. AVAILABILITY: The predicted alternative splice variants are visualized in the SpliceNest database at http://splicenest.molgen.mpg.de PMID- 15117760 TI - Gene-Ontology-based clustering of gene expression data. AB - The expected correlation between genetic co-regulation and affiliation to a common biological process is not necessarily the case when numerical cluster algorithms are applied to gene expression data. GO-Cluster uses the tree structure of the Gene Ontology database as a framework for numerical clustering, and thus allowing a simple visualization of gene expression data at various levels of the ontology tree. AVAILABILITY: The 32-bit Windows application is freely available at http://www.mpibpc.mpg.de/go-cluster/ PMID- 15117761 TI - Immune activation set point during early HIV infection predicts subsequent CD4+ T cell changes independent of viral load. AB - Although generalized T-cell activation is an important factor in chronic HIV disease pathogenesis, its role in primary infection remains poorly defined. To investigate the effect of immune activation on T-cell changes in subjects with early HIV infection, and to test the hypothesis that an immunologic activation "set point" is established early in the natural history of HIV disease, a prospective cohort of acutely infected adults was performed. The median density of CD38 molecules on CD4+ and CD8+ T cells was measured longitudinally in 68 antiretroviral-untreated individuals and 83 antiretroviral-treated individuals. At study entry, T-cell activation was positively associated with viremia, with CD8+ T-cell activation levels increasing exponentially at plasma HIV RNA levels more than 10,000 copies/mL. Among untreated patients, the level of CD8+ T-cell activation varied widely among individuals but often remained stable within a given individual. CD8+ T-cell activation and plasma HIV RNA levels over time were independently associated with the rate of CD4+ T-cell loss in untreated individuals. These data indicate that immunologic activation set point is established early in HIV infection, and that this set point determines the rate at which CD4+ T cells are lost over time. PMID- 15117762 TI - Pleiotropic consequences of Bruton tyrosine kinase deficiency in myeloid lineages lead to poor inflammatory responses. AB - Bruton tyrosine kinase (Btk), a non-receptor-associated tyrosine kinase of the Tec family, appears to participate in many myeloid cell functions. We show that macrophages from X-linked immunodeficient (XID) mice lacking functional Btk cannot generate efficient bursts of reactive oxygen intermediates (ROIs). The induction of apoptotic cell death by inflammatory stimuli is also enhanced in XID macrophages. Phagocytosis of bacterial particles is only marginally affected in them. In vivo, XID mice show reduced severity of inflammatory diseases in models of experimental autoimmune encephalomyelitis (EAE), dextran sulfate sodium (DSS) induced colitis, and carrageenan-induced acute edema. Also, polymorphonuclear neutrophil granulocytes (PMNs) in XID mice show poor ROI and nitric oxide (NO) induction, along with a reduction in PMN recruitment to peritoneal inflammation. XID mice show reduction in PMN numbers in peripheral blood, and their bone marrow shows a reduction in the numbers of both monocytic and granulocytic lineages, extending to the earliest progenitor populations. Thus, Btk is likely to play a significant role at multiple points during the development and functioning of the myeloid lineages, affecting the outcome of many infectious as well as noninfectious inflammatory events in vivo. PMID- 15117763 TI - Risk factors for syngeneic graft-versus-host disease after adult hematopoietic cell transplantation. AB - Syngeneic graft-versus-host disease (sGVHD) has been described after hematopoietic cell transplantation (HCT) but remains poorly defined. We retrospectively reviewed adult syngeneic HCTs at our center (1980-2002) for sGVHD to investigate incidence, morbidity, and risk factors with a primary focus on parity. Among 119 transplantations, there were 21 cases of biopsy-proven sGVHD. The cumulative incidence was 18%, with multiorgan involvement in 6 cases and 1 death. sGVHD was more frequent when the donor was parous (32%) than nulliparous (9%) or male (13%; P =.03) and when the recipient was parous (31%) than nulliparous (7%) or male (13%; P =.02). Other univariable risk factors included older age (P <.01), busulfan/melphalan/thiotepa conditioning (P <.01), interleukin-2 (P =.02), HLA-A26 (P =.03), and more recent transplantation year (P <.01). Overall, risk factors were similar to those described in GVHD. Although an independent effect of parity could not be completely separated from other factors, donor and recipient pregnancy history merits further investigation. PMID- 15117764 TI - Results of the MRC pilot study show autografting for younger patients with chronic lymphocytic leukemia is safe and achieves a high percentage of molecular responses. AB - We have assessed autologous stem cell transplantation after treatment with fludarabine in previously untreated patients with chronic lymphocytic leukemia (CLL). This study is the first to enroll previously untreated patients and follow them prospectively. The initial response rate to fludarabine was 82% (94 of 115 patients). Stem cell mobilization was attempted in 88 patients and was successful in 59 (67%). Overall 65 of 115 patients (56%) entered into the study proceeded to autologous transplantation. The early transplant-related mortality rate was 1.5% (1 of 65 patients). The number of patients in complete remission after transplantation increased from 37% (24 of 65) to 74% (48 of 65), and 26 of 41 patients (63%) who were not in complete remission at the time of their transplantation achieved a complete remission after transplantation. The 5-year overall and disease-free survival rates from transplantation were 77.5% (CI, 57.2%-97.8%) and 51.5% (CI, 33.2%-69.8%), respectively. None of the variables examined at study entry were found to be predictors of either overall or disease free survival. Sixteen of 20 evaluable patients achieved a molecular remission on a polymerase chain reaction (PCR) for immunoglobulin heavy-chain gene rearrangements in the first 6 months following transplantation. Detectable molecular disease by PCR was highly predictive of disease recurrence. It is of concern that 5 of 65 (8%) patients developed posttransplant acute myeloid leukemia/myelodysplastic syndrome. PMID- 15117765 TI - Killing of human immunodeficiency virus-infected primary T-cell blasts by autologous natural killer cells is dependent on the ability of the virus to alter the expression of major histocompatibility complex class I molecules. AB - In the current study, we evaluated whether the capacity of HIV to modulate major histocompatibility complex (MHC) class I molecules has an impact on the ability of autologous natural killer (NK) cells to kill the HIV-infected cells. Analysis of HIV-infected T-cell blasts revealed that the decrease in MHC class I molecules on the infected cell surface was selective. HLA-A and -B were decreased on cells infected with HIV strains that could decrease MHC class I molecules, whereas HLA C and -E remained on the surface. Blocking the interaction between HLA-C and -E and their corresponding inhibitory receptors increased NK cell killing of T-cell blasts infected with HIV strains that reduced MHC class I molecules. Moreover, we demonstrate that NK cells lacking HLA-C and -E inhibitory receptors kill T-cell blasts infected with HIV strains that decrease MHC class I molecules. In contrast, NK cells are incapable of destroying T-cell blasts infected with HIV strains that were unable to reduce MHC class I molecules. These findings suggest that NK cells lacking inhibitory receptors to HLA-C and -E kill HIV-infected CD4+ T cells, and they indicate that the capacity of NK cells to destroy HIV-infected cells depends on the ability of the virus to modulate MHC class I molecules. PMID- 15117766 TI - The implications for health of European Union enlargement. PMID- 15117767 TI - Prevention of respiratory syncytial virus infection in infants. PMID- 15117768 TI - Raised cardiac troponins. PMID- 15117769 TI - Data protection, informed consent, and research. PMID- 15117771 TI - Hepatitis C carriers must be found and treated to avert crisis. PMID- 15117770 TI - NICE advises against caesarean section on demand. PMID- 15117773 TI - Cardiovascular disease threatens developing countries. PMID- 15117775 TI - WHO urges Africa to use more effective malaria medicines. PMID- 15117779 TI - Human Tissue Bill could jeopardise research, scientists warn. PMID- 15117780 TI - Chinese authorities on alert as SARS breaks out again. PMID- 15117781 TI - GMC regrets failure to act on police warning about gynaecologist. PMID- 15117782 TI - AstraZeneca digs in over advertising criticisms. PMID- 15117783 TI - Paediatrician cleared of serious professional misconduct. PMID- 15117784 TI - Living wills will have to specify treatments that patient is refusing. PMID- 15117785 TI - Squeezing academic research into a commercial straitjacket. PMID- 15117786 TI - Creator of Dolly the sheep applies for therapeutic cloning licence. PMID- 15117787 TI - Children are main victims of trafficking in Africa. PMID- 15117789 TI - Drug industry to fight New Zealand's move to ban direct to consumer advertising. PMID- 15117791 TI - Is there a north-south divide in social class inequalities in health in Great Britain? Cross sectional study using data from the 2001 census. AB - OBJECTIVE: To examine individual social class inequalities in self rated general health within and between the constituent countries of Great Britain and the regions of England. DESIGN: Cross sectional study using data from the 2001 national census. SETTING: Great Britain. PARTICIPANTS: Adults aged between 25 and 64 in Great Britain and enumerated in the 2001 population census (n = 25.6 million). MAIN OUTCOME MEASURES: European age standardised rates of self rated general health, for men and women classified by the government social class scheme. RESULTS: In each of the seven social classes, Wales and the North East and North West regions of England had high rates of poor health. There were large social class inequalities in self rated health, with rates of poor health generally increasing from class 1 (higher professional occupations) to class 7 (routine occupations). The size of the health divide varied between regions: the largest rate ratios for routine versus higher professional classes were for Scotland (2.9 for men; 2.8 for women) and London (2.9 for men; 2.4 for women). Women had higher rates of poor health compared to men in the same social class, except in class 6 (semi-routine occupations). CONCLUSIONS: A northwest-southeast divide in social class inequalities existed in Great Britain at the start of the 21st century, with each of the seven social classes having higher rates of poor health in Wales, the North East and North West regions of England than elsewhere. The widest health gap between social classes, however, was in Scotland and London, adding another dimension to the policy debate on resource allocation and targets to tackle the health divide. PMID- 15117792 TI - Reliability of symptoms to determine use of bone scans to identify bone metastases in lung cancer: prospective study. PMID- 15117793 TI - Acute cough in children. PMID- 15117794 TI - Managing nocturia. PMID- 15117795 TI - Pain in the hip and knee. PMID- 15117796 TI - Genetics, race, ethnicity, and health. PMID- 15117797 TI - The logrank test. PMID- 15117798 TI - Response to radiation incidents and radionuclear threats: medical treatment should be given only when safe to do so. PMID- 15117799 TI - Response to radiation incidents and radionuclear threats: other threats may be more serious. PMID- 15117800 TI - Response to radiation incidents and radionuclear threats: renunciation of nuclear weapons could lessen the threat. PMID- 15117801 TI - Penetrating ocular trauma from an exploding microwaved egg. PMID- 15117802 TI - Management of anorexia nervosa revisited: emphasis needs to continue to shift to outpatient care. PMID- 15117803 TI - Management of anorexia nervosa revisited: drug treatment suggestions are questionable. PMID- 15117804 TI - Aspirin induced asthma: clinical relevance of finding was not explained. PMID- 15117805 TI - Aspirin induced asthma: effect size needs to be clarified. PMID- 15117806 TI - E is for equivocal in EBM. PMID- 15117807 TI - Revalidation: swallow hard. PMID- 15117808 TI - Continuing medical education: does "no evidence" trump all? PMID- 15117809 TI - PC cell-derived growth factor (PCDGF/GP88, progranulin) stimulates migration, invasiveness and VEGF expression in breast cancer cells. AB - Metastasis is a multi-step process involved in the progression of breast cancer to a disease with poor prognosis. Growth factor and/or growth factor receptor over- expression have been reported to play an important role in this process. The 88 kDa glycoprotein PC cell-derived growth factor (PCDGF/GP88), also known as progranulin, has been shown to play a major role in breast tumorigenesis by stimulating proliferation, mediating survival and conferring resistance to tamoxifen. In the present paper, the metastatic potential of PCDGF/GP88 was examined in breast cancer. Using MCF-7 cells, we showed that PCDGF/GP88 over expression stimulated anchorage-independent cell growth and accelerated cell migration through matrigel. Similar results were obtained with MCF-7 cells treated exogenously with PCDGF/GP88. Furthermore, gelatin zymograph and immunoblot revealed that matrix metalloprotease-9 was up-regulated by PCDGF/GP88. PCDGF/GP88 stimulated VEGF expression in MCF-7 cells. These results suggest that PCDGF/GP88 could act to promote metastasis and angiogenesis in human breast cancer cells in addition to stimulating their proliferation and survival. PMID- 15117811 TI - Polymorphisms of folate metabolic genes and susceptibility to bladder cancer: a case-control study. AB - Epidemiological studies have shown an association between low folate intake and an increased cancer risk. Major genes involved in folate metabolism include methylene-tetrahydrofolate reductase (MTHFR) and methionine synthase (MS). We investigated joint effects of polymorphisms of the MTHFR (677 C-->T, 1298A-->C) and MS genes (2756 A-->G), dietary folate intake and cigarette smoking on the risk of bladder cancer in a case-control study. The study population consisted of 457 bladder cancer patients and 457 healthy controls, matched to the cases in terms of age, gender and ethnicity. Genotype data were analyzed in a subset of 410 Caucasian cases and 410 controls. Compared with individuals carrying the MTHFR 677 wild-type (CC) and reporting a high folate intake, those carrying the variant genotype (CT or TT) and reporting a low folate intake were at a significantly 3.51-fold increased risk of bladder cancer (95% CI: 1.59-6.52). In contrast, individuals carrying a variant genotype and reporting a high folate intake were at only a 1.39-fold increased risk (95% CI: 0.71-2.70), and those carrying the wild-type and reporting a low folate intake were at only 1.56-fold increased risk (95% CI: 0.82-2.97). The interaction between genetic polymorphisms and folate intake was significant on the multiplicative scale (P = 0.01). When analyzed in the context of smoking status, compared with never smokers with the MTHFR 677 wild-type, the risk increased to 6.56-fold (95% CI: 3.28-13.12) in current smokers carrying the variant genotype. Analyses of the MTHFR 1298, MS 2756 genes revealed similar results. In addition, age at cancer onset in former smokers increased as the proportion of the heteromorphic haplotype in the individual increased (P = 0.005). Our results strongly suggest that polymorphisms of the MTHFR and MS genes act together with low folate intake and smoking to increase bladder cancer risk. These results have important implications for cancer prevention in susceptible populations. PMID- 15117810 TI - Aldehydic DNA lesions in calf thymus DNA and HeLa S3 cells produced by bacterial quinone metabolites of fluoranthene and pyrene. AB - There is increasing concern that compounds formed during the chemical or biological transformation of pollutants in the environment may be more detrimental to human and environmental health than the original pollutant. In this study, two bacterial transformation products of polycyclic aromatic hydrocarbons (PAHs), pyrene-4,5-quinone (P45Q) and fluoranthene-2,3-quinone (F23Q), were evaluated for mutagenicity by measuring aldehydic DNA lesions (ADL) in calf thymus DNA and HeLa S3 cells. Both quinones caused oxidative DNA damage in vitro through a copper-mediated redox cycle and subsequent production of reactive oxygen species (ROS). Hydrogen peroxide and copper were essential for causing oxidative DNA damage and glutathione (GSH) prevented DNA damage from F23Q better than from P45Q. In experiments using HeLa cells, F23Q decreased cell viability, but did not produce measurable levels of ADL or base oxidation. To test the hypothesis that DNA damage was being prevented by conjugation of F23Q with GSH, GSH-depleted cells were treated with both quinones. GSH depletion did not increase the toxicity of F23Q or cause it to oxidize DNA. Treatment of HeLa cells with metal chelators did not decrease F23Q toxicity. It is therefore possible that F23Q affected cell viability through a ROS-independent mechanism, either by conjugation with essential cellular proteins or through cellular or mitochondrial membrane damage, which precluded oxidation of DNA. In contrast, P45Q caused both ADL and base oxidation in cells. Neocuproine reduced the amount of ADL caused by P45Q, indicating that copper was still important for the intracellular generation of damaging oxidants. P45Q is a novel metabolite and its effects on DNA have not been investigated previously. This study exemplifies the importance of considering not only primary environmental pollutants, but also their biologically or chemically generated transformation products. PMID- 15117812 TI - Age-associated changes in the expression pattern of cyclooxygenase-2 and related apoptotic markers in the cancer susceptible region of rat prostate. AB - Senescence-associated changes in the prostate are believed to play an important role in the genesis of prostate cancer. In order to provide further information on how aging increases the prostate susceptibility to cancer, we examined the pattern of cyclooxygenase (COX)-2 expression and the concomitant alterations in prostaglandin E(2) (PGE(2)) synthesis in the prostate glands of 4-, 10-, 50- and 100-week-old Fischer 344 rats. This was carried out in the prostatic areas where hormone-induced tumors arise, namely the periurethral ducts of the dorsolateral prostate (DLP). Age-associated changes were also evaluated for pro- and anti apoptotic factors linked to COX-2 signaling and known to be involved in the normal development of the prostate gland as well as in carcinogenesis. COX-2 expression was increased in the DLP in an age-dependent manner where senescent rats had >3-4-fold higher COX-2 mRNA and protein levels than their juvenile counterparts (P<0.05). The age-related changes in COX-2 were accompanied by a similar up-regulation in the PGE(2) synthesis. Evaluation of mediators of apoptotic signaling showed a significant (P<0.05) decline in the expression levels of the pro-apoptotic BAX (>6-fold) and peroxisome proliferator-activated receptor gamma (>3-fold) and in caspase-3 activity (>2-fold) and an up-regulation of the anti-apoptotic Bcl(2) (>8-fold), PKCalpha (>2-fold) and pAkt (>4-fold) in the 100-week-old rats versus the 4-week-old animals. There was an approximately 15-fold age-dependent decrease in the pro-apoptotic ratio BAX:Bcl(2) and an increase in the anti-apoptotic variable PKCalpha(*)Bcl(2)/BAX in the senescent rats compared with the juvenile ones. These results suggest that increased COX-2 expression can be linked to the decline in the pro-apoptotic signaling in the prostate gland during aging. Subsequently, COX-2 inhibitors can be considered as a promising class of agents to attenuate the increased cell survival and, hence, protect against tumorigenesis in the aging prostate. PMID- 15117813 TI - Fat and K-ras mutations in sporadic colorectal cancer in The Netherlands Cohort Study. AB - Associations between dietary intake of various fats and specific K-ras mutations in colorectal cancer (CRC) were investigated within the framework of The Netherlands Cohort Study on diet and cancer (NLCS). After 7.3 years of follow-up and with exclusion of the first 2.3 years, 448 colon and 160 rectal cancer patients and 2948 subcohort members (55-69 years at baseline) were available for data-analyses. Mutation analysis of the K-ras gene was performed on all archival colon and rectal adenocarcinoma specimens. Case-cohort analyses were used to compute adjusted incidence rate ratios (RR) and 95% confidence intervals (CI) for colon and rectal cancer cases and for K-ras mutation subgroups. The intake of total, saturated and monounsaturated fat was not significantly associated with colon or rectal cancer. High intake of dietary polyunsaturated fat and, specifically, linoleic acid is associated with an increased risk of mutated K-ras colon tumours. The RRs for 1 SD of increase of polyunsaturated fat and linoleic acid were 1.21 (95% CI 1.05-1.41) and 1.22 (95% CI 1.05-1.42), respectively, and similar associations were observed for both G > A transitions and G > T or G > C transversions in the colon. In contrast, no significant associations were observed with rectal cancer risk, overall nor with specific K-ras mutation status. A high intake of polyunsaturated fat, in particular linoleic acid, may be an important dietary risk factor for K-ras mutated colon tumours, possibly by generating G > A transitions or G > T or G > C transversions in the K-ras oncogene. PMID- 15117814 TI - Cell cycle arrest, apoptosis induction and inhibition of nuclear factor kappa B activation in anti-proliferative activity of benzyl isothiocyanate against human pancreatic cancer cells. AB - Benzyl isothiocyanate (BITC), a cruciferous vegetable-derived compound, has been shown to inhibit chemically induced cancer in animal models. Moreover, epidemiological studies have provided compelling evidence to suggest that cruciferous vegetables may be protective against cancer risk. Here, we report that BITC significantly inhibits growth of human pancreatic cancer BxPC-3 cells in a concentration-dependent manner with an IC(50) of approximately 8 micro M, a concentration that can be generated through dietary intake of cruciferous vegetables. Treatment of BxPC-3 cells with growth suppressive concentrations of BITC resulted in G(2)/M phase cell cycle arrest that was associated with a marked decline in protein levels of G(2)/M regulatory proteins including cyclin dependent kinase 1 (Cdk1), cyclin B1 and cell division cycle 25B (Cdc25B). Further, BITC-mediated growth inhibition of BxPC-3 cells correlated with apoptosis induction that was characterized by an increase in Bax/Bcl-2 ratio, cleavage of procaspase-3 and poly(ADP-ribose)polymerase (PARP), and an increase in cytoplasmic histone-associated DNA fragmentation. Interestingly, BITC treatment caused inhibition of nuclear factor kappaB (NF-kappaB) activation, which is constitutively activated in human pancreatic cancer. Western blotting revealed concentration-dependent decrease in NF-kappaB/Rel-p65 protein level in BxPC-3 cells upon exposure to BITC. An increase in protein level of inhibitory subunit kappaB (IkappaBa) in association with reduced serine-32 phosphorylation was also observed in BITC-treated BxPC-3 cells. Consistent with these findings, BITC treatment caused a decrease in nuclear translocation of NF-kappaB as reflected by reduced DNA-binding capacity of NF-kappaB. Furthermore, the protein level of cyclin D1, a transcriptional target of NF-kappaB, was reduced significantly in BITC-treated BxPC-3 cells. To the best of our knowledge, this study is the first published report to implicate suppression of NF-kappaB activation as a potential mechanism for anti-proliferative activity of BITC against human pancreatic cancer cells. PMID- 15117815 TI - Silibinin causes cell cycle arrest and apoptosis in human bladder transitional cell carcinoma cells by regulating CDKI-CDK-cyclin cascade, and caspase 3 and PARP cleavages. AB - Bladder cancer is the fourth and eighth most common cancer in men and women in the USA, respectively. Flavonoid phytochemicals are being studied for both prevention and therapy of various human malignancies including bladder cancer. One such naturally occurring flavonoid is silibinin isolated from milk thistle. Here, we assessed the effect of silibinin on human bladder transitional cell carcinoma (TCC) cell growth, cell cycle modulation and apoptosis induction, and associated molecular alterations, employing two different cell lines representing high-grade invasive tumor (TCC-SUP) and high-grade TCC (T-24) human bladder cancer. Silibinin treatment of these cells resulted in a significant dose- and time-dependent growth inhibition together with a G(1) arrest only at lower doses in TCC-SUP cells but at both lower and higher doses in T-24 cells; higher silibinin dose showed a G(2)/M arrest in TCC-SUP cells. In other studies, silibinin treatment strongly induced the expression of Cip1/p21 and Kip1/p27, but resulted in a decrease in cyclin-dependent kinases (CDKs) and cyclins involved in G(1) progression. Silibinin treatment also showed an increased interaction between cyclin-dependent kinase inhibitors (CDKIs)-CDKs and a decreased CDK kinase activity. Further, the G(2)/M arrest by silibinin in TCC-SUP cells was associated with a decrease in pCdc25c (Ser216), Cdc25c, pCdc2 (Tyr15), Cdc2 and cyclin B1 protein levels. In additional studies, silibinin showed a dose- and a time-dependent apoptotic death only in TCC-SUP cells that was associated with cleaved forms of caspase 3 and poly(ADP-ribose) polymerase. Together, these results suggest that silibinin modulates CDKI-CDK-cyclin cascade and activates caspase 3 causing growth inhibition and apoptotic death of human TCC cells, providing a strong rationale for future studies evaluating preventive and/or intervention strategies for silibinin in bladder cancer pre-clinical models. PMID- 15117816 TI - Thrombosis and neointima formation in vein grafts are inhibited by locally applied aspirin through endothelial protection. AB - Vein graft failure within the first month after bypass surgery is largely because of thrombosis. However, systemic study of thrombus formation in vein grafts is still lacking, and few effective techniques are available to prevent this event. Herein, we analyzed the kinetics of thrombosis and tested the effectiveness of locally applied aspirin on prevention of the disease in a mouse model. En face analysis of vein grafts revealed that 67+/-12% and 54+/-17% of the surface areas were covered by microthrombi at 1 and 3 days, respectively. Thrombus generation was also identified by labeling of platelets and fibrin, which occurred in 35 grafts examined at 1 and 3 days and 1, 2, 4, and 8 weeks. In a fifth of grafts, the thrombus occluded the vessel lumen by > or =1/4. Furthermore, a significant loss of endothelial cells was evidenced by beta-gal staining for vein grafts in transgenic mice expressing LacZ gene controlled by TIE2-endothelial specific gene promoter. Following thrombosis, neointimal lesions were significantly increased by 4-fold 2 weeks after the operation. When vein grafts were treated locally with aspirin in pluronic gel-127, the thrombus area was significantly reduced (P<0.005) at 1, 4, and 8 weeks. Interestingly, neointimal lesions were markedly reduced in the local, but not oral, aspirin-treated group at 4 and 8 weeks by 50% to 70% (P<0.005). The mechanism of reduced lesions by locally applied aspirin involved the protection of vein graft endothelium. Thus, we provide strong evidence that thrombus formation occurs before the development of neointimal lesions in vein grafts and that local aspirin treatment successfully reduces vein graft arteriosclerosis through endothelial protection, resulting in reduction of thrombosis. PMID- 15117817 TI - Enhanced contractility of renal afferent arterioles from angiotensin-infused rabbits: roles of oxidative stress, thromboxane prostanoid receptors, and endothelium. AB - We tested the hypothesis that cyclooxygenase (COX), thromboxane A2 synthase (TxA2 S), thromboxane prostanoid receptors (TP-Rs), or superoxide anion (O2-) mediates enhanced contractions of renal afferent arterioles (Aff) of angiotensin II (Ang II)-infused rabbits. Rabbits were infused with vehicle (sham), Ang II 60 ng x kg( 1) x min(-1) (Ang II 60) or 200 ng x kg(-1) x min(-1) (Ang II 200). There was a selective enhanced vasoconstriction of Affs from Ang II 60 rabbits to Ang II (Deltadiameter-78+/-8% versus -43+/-9%; P<0.01) that was normalized by a TP-R antagonist but not by a superoxide dismutase (SOD) mimetic. Affs from Ang II 200 rabbits had increased (P<0.01) mRNA for COX-2 and enhanced vasoconstriction to Ang II, U-46 619 (TP-R mimetic), and endothelin-1 that was normalized by ifetroban plus tempol together. Endothelium removal enhanced Ang II responses of Affs from sham rabbits but blunted responses from Ang II 200 rabbits and abolished responses to ifetroban. Affs from Ang II 200 rabbits had an endothelium dependent contraction factor (EDCF) response to that was blunted (P<0.001) by a SOD mimetic or antagonists of COX-1 or TxA2-S but normalized by antagonists of COX-2 or TP-R. Thus, enhanced Ang II responses in Affs from rabbits infused with slow pressor Ang II are mediated independently by O2- in the vascular smooth muscle cells and by an EDCF that is principally a vasoconstrictor prostaglandin generated by COX-2 >-1 activating TP-Rs, whereas enhanced responses in rabbits infused with a lower Ang II dose are dependent on TP-R but not O2-. PMID- 15117818 TI - Endothelin-1-dependent nuclear factor of activated T lymphocyte signaling associates with transcriptional coactivator p300 in the activation of the B cell leukemia-2 promoter in cardiac myocytes. AB - Endothelin-1 (ET-1) is a potent survival factor that protects cardiac myocytes from apoptosis. ET-1 induces cardiac gene transcription and protein expression of antiapoptotic B cell leukemia-2 (bcl-2) in a calcineurin-dependent manner. A cellular target of adenovirus early region 1A (E1A) oncoprotein, p300 also activates bcl-2 transcription in cardiac myocytes and is required for their survival. p300 acts as a calcineurin-regulated nuclear factors of activated T lymphocytes (NFATc), downstream targets of calcineurin. In addition, the bcl-2 promoter contains multiple NFAT consensus sequences. These findings prompted us to investigate the role of NFATc in ET-1-dependent and p300-dependent bcl-2 transcription in cardiac myocytes. In primary cardiac myocytes prepared from neonatal rats, mutation of 2 NFAT sites within the bcl-2 promoter completely abolished the ET-1- and p300-induced increases in the activity of this promoter. We show here that p300 markedly potentiates the binding of NFATc1 to the bcl-2 NFAT element by interacting with NFATc1 in an E1A-dependent manner. On the other hand, stimulation of cardiac myocytes with ET-1 causes nuclear translocation of NFATc1, which interacts with p300 and increases DNA binding. Expression of E1A did not change the cardiac nuclear localization of NFATc1 but blocked its interaction with p300, DNA binding, and bcl-2 promoter activation. These findings suggest that ET-1-dependent NFATc signaling associates with p300 in the transactivation of bcl-2 gene in cardiac myocytes. PMID- 15117819 TI - Chromosome 1q21.1 contiguous gene deletion is associated with congenital heart disease. AB - Congenital heart disease (CHD), comprising structural or functional abnormalities present at birth, is the most common birth defect in humans. Reduced expression of connexin40 (Cx40) has been found in association with atrial fibrillation, and deletion of Cx40 in a mouse model causes various structural heart abnormalities in 18% of heterozygotes. We screened 505 unrelated CHD cases for deletions or duplications of the Cx40 gene (GJA5) by real-time quantitative PCR, in order to determine whether altered copy number of this gene may be associated with a cardiac phenotype in humans. Dosage of Cx40 flanking genes (ACPL1 and Cx50 gene, GJA8) was determined by real-time PCR for all apparent positive cases. In total, 3 cases were found to carry deletions on chromosome 1q21.1 spanning ACPL1, Cx40, and Cx50 genes. Absence of heterozygosity was observed in all 3 index cases over a 1.5- to 3-Mb region. Samples from the parents of two cases were obtained, and microsatellites across 1q21.1 were genotyped. One of the apparently unaffected parents was found to carry this deletion. All 3 index cases presented with obstruction of the aortic arch as the common structural cardiac malformation, and had no consistent dysmorphic features. Genotyping of 520 unrelated normal controls for this deletion was negative. We hypothesize that this 1q21.1 multigene deletion is associated with a range of cardiac defects, with anomalies of the aortic arch being a particular feature. PMID- 15117820 TI - Paired-related homeobox gene Prx1 is required for pulmonary vascular development. AB - Herein, we show that the paired-related homeobox gene, Prx1, is required for lung vascularization. Initial studies revealed that Prx1 localizes to differentiating endothelial cells (ECs) within the fetal lung mesenchyme, and later within ECs forming vascular networks. To begin to determine whether Prx1 promotes EC differentiation, fetal lung mesodermal cells were transfected with full-length Prx1 cDNA, resulting in their morphological transformation to an endothelial-like phenotype. In addition, Prx1-transformed cells acquired the ability to form vascular networks on Matrigel. Thus, Prx1 might function by promoting pulmonary EC differentiation within the fetal lung mesoderm, as well as their subsequent incorporation into vascular networks. To understand how Prx1 participates in network formation, we focused on tenascin-C (TN-C), an extracellular matrix (ECM) protein induced by Prx1. Immunocytochemistry/histochemistry showed that a TN-C rich ECM surrounds Prx1-positive pulmonary vascular networks both in vivo and in tissue culture. Furthermore, antibody-blocking studies showed that TN-C is required for Prx1-dependent vascular network formation on Matrigel. Finally, to determine whether these results were relevant in vivo, we examined newborn Prx1 wild-type (+/+) and Prx1-null (-/-) mice and showed that Prx1 is critical for expression of TN-C and lung vascularization. These studies provide a framework to understand how Prx1 controls EC differentiation and their subsequent incorporation into functional pulmonary vascular networks. PMID- 15117821 TI - Endothelial lipase promotes the catabolism of ApoB-containing lipoproteins. AB - Endothelial lipase (EL) has been found to be a key enzyme in high-density lipoprotein (HDL) metabolism in mice, leading to the concept that inhibition of EL could be a novel strategy for raising HDL cholesterol levels. However, mice are "HDL animals" and the effect of EL on atherogenic apoB-containing lipoproteins has not been elucidated. We previously found that EL is capable of hydrolyzing very low-density lipoprotein (VLDL) and LDL lipids ex vivo. To investigate the role of EL in the metabolism of apoB-containing lipoproteins in vivo, we expressed human EL in three mouse models of elevated apoB-containing lipoproteins: apoE-deficient, LDL receptor-deficient, and human apoB transgenic mice. Unexpectedly, hepatic expression of EL resulted in markedly decreased levels of VLDL/LDL cholesterol, phospholipid, and apoB accompanied by significantly increased LDL apolipoprotein and phospholipid catabolism. To determine whether lipolytic activity is required for this effect, we also expressed a catalytically inactive form of human EL (ELS149A); unexpectedly, expression of ELS149A did not lower and in fact increased plasma lipids. Coexpression and coimmunoprecipitation studies suggested that catalytically inactive ELS149A inhibits endogenous mouse EL, accounting for the increased lipid levels. We conclude that (1) in addition to its known effects on HDL metabolism, EL influences the metabolism of apoB-containing particles; (2) catalytic activity of EL is required for its effects on apoB-containing lipoproteins; and (3) overexpressed catalytically inactive EL inhibits endogenous mouse EL, resulting in increased levels of plasma lipids. In light of these results, inhibition of EL has the potential to raise levels of atherogenic lipoproteins in addition to HDL C levels. PMID- 15117822 TI - Proangiogenic action of thyroid hormone is fibroblast growth factor-dependent and is initiated at the cell surface. AB - The effects of thyroid hormone analogues on modulation of angiogenesis have been studied in the chick chorioallantoic membrane model. Generation of new blood vessels from existing vessels was increased 3-fold by either l-thyroxine (T4; 10( 7) mol/L) or 3,5,3'-triiodo-l-thyronine (10(-9) mol/L). T4-agarose reproduced the effects of T4, and tetraiodothyroacetic acid (tetrac) inhibited the effects of both T4 and T4-agarose. Tetrac itself was inactive and is known to block actions of T4 on signal transduction that are initiated at the plasma membrane. T4 and basic fibroblast growth factor (FGF2) were comparably effective as inducers of angiogenesis. Low concentrations of FGF2 combined with submaximal concentrations of T4 produced an additive angiogenic response. Anti-FGF2 inhibited the angiogenic effect of T4. The proangiogenic effects of T4 and FGF2 were blocked by PD 98059, a mitogen-activated protein kinase (MAPK) pathway inhibitor. Endothelial cells (ECV304) treated with T4 or FGF2 for 15 minutes demonstrated activation of MAPK, an effect inhibited by PD 98059 and the protein kinase C inhibitor CGP41251. Reverse transcription-polymerase chain reaction of RNA extracted from endothelial cells treated with T4 revealed increased abundance of FGF2 transcript at 6 to 48 hours, and after 72 hours, the medium of treated cells showed increased FGF2 content, an effect inhibited by PD 98059. Thus, thyroid hormone is shown to be a proangiogenic factor. This action, initiated at the plasma membrane, is MAPK dependent and mediated by FGF2. PMID- 15117823 TI - Increased vulnerability to atrial fibrillation in transgenic mice with selective atrial fibrosis caused by overexpression of TGF-beta1. AB - Studies on patients and large animal models suggest the importance of atrial fibrosis in the development of atrial fibrillation (AF). To investigate whether increased fibrosis is sufficient to produce a substrate for AF, we have studied cardiac electrophysiology (EP) and inducibility of atrial arrhythmias in MHC TGFcys33ser transgenic mice (Tx), which have increased fibrosis in the atrium but not in the ventricles. In anesthetized mice, wild-type (Wt) and Tx did not show significant differences in surface ECG parameters. With transesophageal atrial pacing, no significant differences were observed in EP parameters, except for a significant decrease in corrected sinus node recovery time in Tx mice. Burst pacing induced AF in 14 of 29 Tx mice, whereas AF was not induced in Wt littermates (P<0.01). In Langendorff perfused hearts, atrial conduction was studied using a 16-electrode array. Epicardial conduction velocity was significantly decreased in the Tx RA compared with the Wt RA. In the Tx LA, conduction velocity was not significantly different from Wt, but conduction was more heterogeneous. Action potential characteristics recorded with intracellular microelectrodes did not reveal differences between Wt and Tx mice in either atrium. Thus, in this transgenic mouse model, selective atrial fibrosis is sufficient to increase AF inducibility. PMID- 15117824 TI - Thioredoxin-2 inhibits mitochondria-located ASK1-mediated apoptosis in a JNK independent manner. AB - Apoptosis signal-regulating kinase 1 (ASK1) mediates cytokines and oxidative stress (ROS)-induced apoptosis in a mitochondria-dependent pathway. However, the underlying mechanism has not been defined. In this study, we show that ASK1 is localized in both cytoplasm and mitochondria of endothelial cells (ECs) where it binds to cytosolic (Trx1) and mitochondrial thioredoxin (Trx2), respectively. Cys 250 and Cys-30 in the N-terminal domain of ASK1 are critical for binding of Trx1 and Trx2, respectively. Mutation of ASK1 at C250 enhanced ASK1-induced JNK activation and apoptosis, whereas mutation of ASK1 at C30 specifically increased ASK1-induced apoptosis without effects on JNK activation. We further show that a JNK-specific inhibitor SP600125 completely blocks TNF induced JNK activation, Bid cleavage, and Bax mitochondrial translocation, but only partially inhibits cytochrome c release and EC death, suggesting that TNF induces both JNK-dependent and JNK-independent apoptotic pathways in EC. Mitochondria-specific expression of a constitutively active ASK1 strongly induces EC apoptosis without JNK activation, Bid cleavage, and Bax mitochondrial translocation. These data suggest that mitochondrial ASK1 mediates a JNK-independent apoptotic pathway induced by TNF. To determine the role of Trx2 in regulation of mitochondrial ASK1 activity, we show that overexpression of Trx2 inhibits ASK1-induced apoptosis without effects on ASK1-induced JNK activation. Moreover, specific knockdown of Trx2 in EC increases TNF/ASK1-induced cytochrome c release and cell death without increase in JNK activation, Bid cleavage, and Bax translocation. Our data suggest that ASK1 in cytoplasm and mitochondria mediate distinct apoptotic pathways induced by TNF, and Trx1 and Trx2 cooperatively inhibit ASK1 activities. PMID- 15117825 TI - PPARgamma agonists ameliorate endothelial cell activation via inhibition of diacylglycerol-protein kinase C signaling pathway: role of diacylglycerol kinase. AB - Subject- Peroxisome proliferator-activated receptor (PPAR)-gamma agonists are emerging as potential protectors against inflammatory cardiovascular diseases including atherosclerosis and diabetic complications. However, their molecular mechanism of action within vasculature remains unclear. We report here that PPARgamma agonists, thiazolidinedione class drugs (TZDs), or 15-deoxy-Delta12,14 prostaglandin J2 (15d-PGJ2) were capable of activating diacylglycerol (DAG) kinase (DGK), resulting in attenuation of DAG levels and inhibition of protein kinase C (PKC) activation. The PPARgamma agonist-induced DGK was completely blocked by a dominant-negative mutant of PPARgamma, indicating an essential receptor-dependent action. Importantly, the suppression of DAG-PKC signaling pathway was functional linkage to the anti-inflammatory properties of PPARgamma agonists in endothelial cells (EC), characterized by the inhibition of proinflammatory adhesion molecule expression and adherence of monocytes to the activated EC induced by high glucose. These findings thus demonstrate a novel molecular action of PPARgamma agonists to suppress the DAG-PKC signaling pathway via upregulation of an endogenous attenuator, DGK. PMID- 15117826 TI - Chronic production of angiotensin IV in the brain leads to hypertension that is reversible with an angiotensin II AT1 receptor antagonist. AB - Angiotensin IV (Ang IV) is a metabolite of the potent vasoconstrictor angiotensin II (Ang II). Because specific binding sites for this peptide have been reported in numerous tissues including the brain, it has been suggested that a specific Ang IV receptor (AT4) might exist. Bolus injection of Ang IV in brain ventricles has been implicated in learning, memory, and localized vasodilatation. However, the functions of Ang IV in a physiological context are still unknown. In this study, we generated a transgenic (TG) mouse model that chronically releases Ang IV peptide specifically in the brain. TG mice were found to be hypertensive by the tail-cuff method as compared with control littermates. Treatment with the angiotensin-converting enzyme inhibitor captopril had no effect on blood pressure, but surprisingly treatment with the Ang II AT1 receptor antagonist candesartan normalized the blood pressure despite the fact that the levels of Ang IV in the brains of TG mice were only 4-fold elevated over the normal endogenous level of Ang peptides. Calcium mobilization assays performed on cultured CHO cells chronically transfected with the AT1 receptor confirm that low-dose Ang IV can mobilize calcium via the AT1 receptor only in the presence of Ang II, consistent with an allosteric mechanism. These results suggest that chronic elevation of Ang IV in the brain can induce hypertension that can be treated with angiotensin II AT1 receptor antagonists. PMID- 15117827 TI - The mouse through the looking glass: a new door into the pathophysiology of pulmonary hypertension. PMID- 15117828 TI - Myocardial hibernation: a double-edged sword. PMID- 15117829 TI - Imaging microdomain Ca2+ in muscle cells. AB - Ca2+ ions passing through a single or a cluster of Ca2+-permeable channels create microscopic, short-lived Ca2+ gradients that constitute the building blocks of cellular Ca2+ signaling. Over the last decade, imaging microdomain Ca2+ in muscle cells has unveiled the exquisite spatial and temporal architecture of intracellular Ca2+ dynamics and has reshaped our understanding of Ca2+ signaling mechanisms. Major advances include the visualization of "Ca2+ sparks" as the elementary events of Ca2+ release from the sarcoplasmic reticulum (SR), "Ca2+ sparklets" produced by openings of single Ca2+-permeable channels, miniature Ca2+ transients in single mitochondria ("marks"), and SR luminal Ca2+ depletion transients ("scraps"). As a model system, a cardiac myocyte contains a 3 dimensional grid of 104 spark ignition sites, stochastic activation of which summates into global Ca2+ transients. Tracking intermolecular coupling between single L-type Ca2+ channels and Ca2+ sparks has provided direct evidence validating the local control theory of Ca2+-induced Ca2+ release in the heart. In vascular smooth muscle myocytes, Ca2+ can paradoxically signal both vessel constriction (by global Ca2+ transients) and relaxation (by subsurface Ca2+ sparks). These findings shed new light on the origin of Ca2+ signaling efficiency, specificity, and versatility. In addition, microdomain Ca2+ imaging offers a novel modality that complements electrophysiological approaches in characterizing Ca2+ channels in intact cells. PMID- 15117830 TI - The dystrophin glycoprotein complex: signaling strength and integrity for the sarcolemma. AB - The dystrophin glycoprotein complex (DGC) is a specialization of cardiac and skeletal muscle membrane. This large multicomponent complex has both mechanical stabilizing and signaling roles in mediating interactions between the cytoskeleton, membrane, and extracellular matrix. Dystrophin, the protein product of the Duchenne and X-linked dilated cardiomyopathy locus, links cytoskeletal and membrane elements. Mutations in additional DGC genes, the sarcoglycans, also lead to cardiomyopathy and muscular dystrophy. Animal models of DGC mutants have shown that destabilization of the DGC leads to membrane fragility and loss of membrane integrity, resulting in degeneration of skeletal muscle and cardiomyocytes. Vascular reactivity is altered in response to primary degeneration in striated myocytes and arises from a vascular smooth muscle cell-extrinsic mechanism. PMID- 15117831 TI - Bone marrow-derived cells and vascular growth. PMID- 15117832 TI - Surgical reconstruction of occluded pulmonary arteries in patients with congenital heart disease: effects on pulmonary artery growth. AB - BACKGROUND: This study was undertaken to determine outcomes and best strategies for treatment of occluded pulmonary arteries in patients with congenital heart disease. METHODS AND RESULTS: Between 1998 and 2002, occlusion of a previously patent pulmonary artery was established in 23 patients. Data were obtained retrospectively. Diagnoses were pulmonary atresia and ventricular septal defect in 11, tetralogy of Fallot in 6, and other forms of pulmonary stenosis or atresia in 6. Median age and weight at diagnosis were 9 years (range, 6 days to 43 years) and 24 kg (range, 2.6 to 60 kg). Fourteen patients had had a previous surgery. The occluded pulmonary artery was visualized at angiography by wedge injection or injection into the collateral circulation. The left pulmonary artery was occluded in 20 patients and the right pulmonary artery in 3. Criteria for reconstruction were estimated duration of occlusion <6 months and ratio of occluded to contralateral artery >0.2. Twelve patients fulfilled these criteria and underwent pulmonary artery reconstruction at a mean interval of 2 months (range, 6 days to 6 months) from evidence of occlusion. Six patients had pericardial patch reconstruction, 3 terminoterminal anastomosis, 2 thrombectomy, and 1 a Blalock Taussig shunt. There was 1 late death. At a median follow-up of 4 years (2 months to 5 years), all patients underwent cardiac catheterization: in 8 patients the reconstructed artery was patent, in 3 reoccluded. Hypoplasia of the occluded artery was reversed in 6 patients. CONCLUSIONS: Our data show that in selected patients, reconstruction of an occluded pulmonary artery can restore pulmonary vascularization and reverse hypoplasia. Strict surveillance is mandatory to prevent pulmonary artery loss. PMID- 15117833 TI - Independent factors associated with outcomes of parachute mitral valve in 84 patients. AB - BACKGROUND: Parachute mitral valve (PMV) is defined as a unifocal attachment of the mitral valve chordae to a single or dominant papillary muscle and may cause subvalvar obstruction. We sought to determine factors associated with outcomes. METHODS AND RESULTS: Patients (n=84; 64% male) who presented between 1977 and 2001 at a median age of 3 days (range, birth to 5.4 years) were assessed with PMV (without atrioventricular septal defect). Associated cardiac anomalies in 99% included aortic coarctation in 68%, atrial septal defect in 54%, ventricular septal defect in 46%, aortic valve stenosis in 32%, subaortic stenosis in 20%, and left ventricular hypoplasia in 19%, with complex anomalies in 14%. Noncardiac anomalies were noted in 32%. Survival (n=18 deaths) was 82% at 1 year and 79% at 10 years, with independent risk factors including left ventricular hypoplasia (P<0.001) and atrial septal defect (P<0.003). Freedom from surgical mitral valvotomy (n=11 patients) was 95% at age 6 months and 80% at 10 years, with independent risk factors including the absence of aortic coarctation (P<0.02) and the presence of subaortic stenosis (P<0.04). There was no significant increase in mean gradient of the PMV over time, but higher gradient was independently associated with the presence of supravalvar mitral stenosis (P<0.001), absence of atrial septal defect (P<0.04), presence of ventricular septal defect (P<0.02), and subsequent mitral valvotomy (P<0.01). CONCLUSIONS: Outcomes for patients with PMV are dependent on the spectrum of associated cardiac lesions. The degree of mitral valve obstruction remains stable, and the majority will not require valvotomy. PMID- 15117834 TI - Magnetic resonance low-dose dobutamine test is superior to SCAR quantification for the prediction of functional recovery. AB - BACKGROUND: Low-dose dobutamine challenge (DSMR) by MRI was compared with delayed enhancement imaging with Gd-DTPA (SCAR) as a predictor of improvement of wall motion after revascularization (RECOVERY). METHODS AND RESULTS: In 29 patients with coronary artery disease (68+/-7 years of age, 2 women, 32+/-8% ejection fraction), wall motion was evaluated semiquantitatively by MRI before and 3 months after revascularization. SCAR and DSMR were performed before revascularization. The transmural extent of scar was assessed semiquantitatively. Binary prediction of RECOVERY was performed by logistic regression in 288 segments with wall motion abnormalities at rest. Receiver operating characteristic-area under curve (AUC) statistics were used to compare different models. Low-dose DSMR (AUC 0.838) was superior to SCAR (AUC 0.728) in predicting RECOVERY. SCAR did not improve accuracy of prediction by DSMR. Subgroup analysis showed superiority of DSMR for 1% to 74% transmural extent of infarction. CONCLUSIONS: Low-dose DSMR is superior to SCAR in predicting RECOVERY. This advantage is largest in segments with a delayed enhancement of 1% to 74%. PMID- 15117835 TI - Angiotensin II type 2 receptor-mediated vasodilation in human coronary microarteries. AB - BACKGROUND: Angiotensin (Ang) II type 2 (AT2) receptor stimulation results in coronary vasodilation in the rat heart. In contrast, AT2 receptor-mediated vasodilation could not be observed in large human coronary arteries. We studied Ang II-induced vasodilation of human coronary microarteries (HCMAs). METHODS AND RESULTS: HCMAs (diameter, 160 to 500 microm) were obtained from 49 heart valve donors (age, 3 to 65 years). Ang II constricted HCMAs, mounted in Mulvany myographs, in a concentration-dependent manner (pEC50, 8.6+/-0.2; maximal effect [E(max)], 79+/-13% of the contraction to 100 mmol/L K+). The Ang II type 1 receptor antagonist irbesartan prevented this vasoconstriction, whereas the AT2 receptor antagonist PD123319 increased E(max) to 97+/-14% (P<0.05). The increase in E(max) was larger in older donors (correlation DeltaE(max) versus age, r=0.47, P<0.05). The PD123319-induced potentiation was not observed in the presence of the NO synthase inhibitor L-NAME, the bradykinin type 2 (B2) receptor antagonist Hoe140, or after removal of the endothelium. Ang II relaxed U46619-preconstricted HCMAs in the presence of irbesartan by maximally 49+/-16%, and PD123319 prevented this relaxation. Finally, radioligand binding studies and reverse transcription polymerase chain reaction confirmed the expression of AT2 receptors in HCMAs. CONCLUSIONS: AT2 receptor-mediated vasodilation in the human heart appears to be limited to coronary microarteries and is mediated by B2 receptors and NO. Most likely, AT2 receptors are located on endothelial cells, and their contribution increases with age. PMID- 15117836 TI - Increased reactive oxygen species in rostral ventrolateral medulla contribute to neural mechanisms of hypertension in stroke-prone spontaneously hypertensive rats. AB - BACKGROUND: Oxidative stress increases in hypertension. The aim of this study was to determine whether reactive oxygen species (ROS) are increased in the rostral ventrolateral medulla (RVLM) in the brainstem, where the vasomotor center is located, in stroke-prone spontaneously hypertensive rats (SHRSP), and, if so, to determine whether the increased ROS contribute to neural mechanisms of hypertension in SHRSP. METHODS AND RESULTS: We measured ROS levels in the RVLM of SHRSP and compared them with those in Wistar-Kyoto rats (WKY). Thiobarbituric acid-reactive substances were increased in SHRSP compared with WKY. ROS were measured by electron spin resonance (ESR) spectroscopy. The ESR signal decay rate in the RVLM of SHRSP was significantly increased compared with that in WKY, and this increase was abolished by dimethylthiourea (a hydroxyl radical scavenger). The increased ESR signal decay rate was reduced to the same extent in the presence of desferrioxamine, catalase, and Tiron, indicating that hydroxyl radicals are derived from superoxide anions and hydrogen peroxide. In addition, total superoxide dismutase (SOD) activity in the RVLM was decreased in SHRSP compared with WKY. Furthermore, bilateral microinjection of tempol into the RVLM decreased blood pressure in SHRSP but not in WKY, and MnSOD overexpression in the RVLM of SHRSP decreased blood pressure and inhibited sympathetic nerve activity. CONCLUSIONS: These results suggest that superoxide anions in the RVLM, which generate hydroxyl radicals, are increased in SHRSP and contribute to the neural mechanisms of hypertension in SHRSP. PMID- 15117837 TI - Intramural virtual electrodes in ventricular wall: effects on epicardial polarizations. AB - BACKGROUND: Intramural virtual electrodes (IVEs) are believed to play an important role in defibrillation, but their existence in intact myocardium remains unproven. Here, IVEs were detected by use of optical recordings of shock induced transmembrane potential (V(m)) changes (DeltaV(m)) measured from the intact epicardial heart surface. METHODS AND RESULTS: To detect IVEs, isolated porcine left ventricles were sequentially stained with a V(m)-sensitive dye by 2 methods: (1) surface staining (SS) and (2) global staining (GS) via coronary perfusion. Shocks (2 to 50 V/cm) were applied across the ventricular wall in an epicardial-to-endocardial direction during the action potential plateau via transparent mesh electrodes, and shock-induced DeltaV(m) were measured optically from the same epicardial locations after SS and GS. Optical recordings revealed significant differences between DeltaV(m) of 2 types that became more prominent with increasing shock strength: (1) for weak shocks, SS-DeltaV(m) were larger and faster than GS-DeltaV(m); (2) for intermediate shocks, cathodal GS-DeltaV(m) became multiphasic, whereas SS-DeltaV(m) remained monophasic; and (3) for strong shocks, cathodal GS-DeltaV(m) became uniformly negative, whereas SS-DeltaV(m) typically remained positive. The radical differences in the shape and polarity of SS and GS polarizations can be explained by the contribution of subepicardial IVEs to optical signals. Histological examination revealed a dense network of collagen septa in the subepicardium, which could form the IVE substrate. CONCLUSIONS: Intramural virtual electrodes are reflected in optical measurements of shock-induced DeltaV(m) on the intact epicardial surface. These IVEs could be a result of microscopic resistive discontinuities formed by collagen septa. PMID- 15117838 TI - Glucocorticoid treatment prevents progressive myocardial dysfunction resulting from experimental coronary microembolization. AB - BACKGROUND: The frequency and importance of microembolization in patients with acute coronary syndromes and during coronary interventions have recently been appreciated. Experimental microembolization induces immediate ischemic dysfunction, which recovers within minutes. Subsequently, progressive contractile dysfunction develops over several hours and is not associated with reduced regional myocardial blood flow (perfusion-contraction mismatch) but rather with a local inflammatory reaction. We have now studied the effect of antiinflammatory glucocorticoid treatment on this progressive contractile dysfunction. METHODS AND RESULTS: Microembolization was induced by injecting microspheres (42-microm diameter) into the left circumflex coronary artery. Anesthetized dogs were followed up for 8 hours and received placebo (n=7) or methylprednisolone 30 mg/kg IV either 30 minutes before (n=7) or 30 minutes after (n=5) microembolization. In addition, chronically instrumented dogs received either placebo (n=4) or methylprednisolone (n=4) 30 minutes after microembolization and were followed up for 1 week. In acute placebo dogs, posterior systolic wall thickening was decreased from 20.0+/-2.1% (mean+/-SEM) at baseline to 5.8+/-0.6% at 8 hours after microembolization. Methylprednisolone prevented the progressive myocardial dysfunction. Increased leukocyte infiltration in the embolized myocardium was prevented only when methylprednisolone was given before microembolization. In chronic placebo dogs, progressive dysfunction recovered from 5.0+/-0.7% at 4 to 6 hours after microembolization back to baseline (19.1+/-1.6%) within 5 days. Again, methylprednisolone prevented the progressive myocardial dysfunction. CONCLUSIONS: Methylprednisolone, even when given after microembolization, prevents progressive contractile dysfunction. PMID- 15117839 TI - Independent association between plasma leptin and C-reactive protein in healthy humans. AB - BACKGROUND: C-reactive protein (CRP) is synthesized from the liver and is regulated by cytokines, especially interleukin-6. Leptin, the adipocyte-derived protein product of the ob gene, is related to amount of body fat. The long form of the leptin receptor resembles cytokine receptors, which include the interleukin-6 receptor. Both leptin and CRP may be increased in women, in obesity, and in inflammation, and both have been linked to cardiovascular pathophysiological processes and increased cardiovascular risk. We tested the hypothesis that leptin is associated with CRP levels independently of the influences of gender, body mass index (BMI), waist-to-hip ratio, and other variables. METHODS AND RESULTS: We studied 100 healthy volunteers (48 men, and 52 women). For all subjects, leptin was independently associated with CRP after adjustment for age, gender, BMI, waist-to-hip ratio, smoking, and alcohol consumption (F=12.39, P=0.0007). There was a strong and significant positive relationship between leptin and CRP in both women (R=0.61, P<0.0001) and men (R=0.55, P<0.0001) considered separately. The association between leptin and CRP was significant even after adjustment for age, BMI, waist-to-hip ratio, smoking, and alcohol consumption in women (F=7.13, P=0.01) and men (F=5.69, P=0.02). When only subjects with BMI <25 kg/m2 were considered (n=47), CRP was not linked to BMI (R=0.02, P=0.96), but a significant association between leptin and CRP was still evident (R=0.55, P<0.0001). CONCLUSIONS: Leptin and CRP levels are independently associated in normal humans, providing further evidence linking metabolic and inflammatory cardiovascular disease mechanisms. PMID- 15117840 TI - Ghrelin inhibits proinflammatory responses and nuclear factor-kappaB activation in human endothelial cells. AB - BACKGROUND: Ghrelin is a novel growth hormone-releasing peptide that has been shown to improve cachexia in heart failure and cancer and to ameliorate the hemodynamic and metabolic disturbances in septic shock. Because cytokine-induced inflammation is critical in these pathological states and because the growth hormone secretagogue receptor has been identified in blood vessels, we examined whether ghrelin inhibits proinflammatory responses in human endothelial cells in vitro and after administration of endotoxin to rats in vivo. METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) were treated with or without tumor necrosis factor-alpha (TNF-alpha), and induction of proinflammatory cytokines and mononuclear cell adhesion were determined. Ghrelin (0.1 to 1000 ng/mL) inhibited both basal and TNF-alpha-induced cytokine release and mononuclear cell binding. Intravenous administration of ghrelin also inhibited endotoxin-induced proinflammatory cytokine production in rats in vivo. Ghrelin inhibited H2O2-induced cytokine release in HUVECs, suggesting that the peptide blocks redox-mediated cellular signaling. Moreover, ghrelin inhibited basal and TNF-alpha-induced activation of nuclear factor-kappaB. Des-acyl ghrelin had no effect on TNF-alpha-induced cytokine production in HUVECs, suggesting that the antiinflammatory effects of ghrelin require interaction with endothelial growth hormone secretagogue receptors. CONCLUSIONS: Ghrelin inhibits proinflammatory cytokine production, mononuclear cell binding, and nuclear factor-kappaB activation in human endothelial cells in vitro and endotoxin-induced cytokine production in vivo. These novel antiinflammatory actions of ghrelin suggest that the peptide could play a modulatory role in atherosclerosis, especially in obese patients, in whom ghrelin levels are reduced. PMID- 15117841 TI - Angiotensin type 1 receptor blockers induce peroxisome proliferator-activated receptor-gamma activity. AB - BACKGROUND: Angiotensin type 1 receptor (AT(1)R) blockers (ARB) have been shown to reduce the incidence of type 2 diabetes mellitus by an unknown molecular mechanism. The peroxisome proliferator-activated receptor-gamma (PPARgamma) is the central regulator of insulin and glucose metabolism improving insulin sensitivity. We investigated the regulation of PPARgamma function by ARBs. METHODS AND RESULTS: The ARBs irbesartan and telmisartan (10 micromol/L) potently enhanced PPARgamma-dependent 3T3-L1 adipocyte differentiation associated with a significant increase in mRNA expression of the adipogenic marker gene adipose protein 2 (aP2), as measured by quantitative real-time polymerase chain reaction (irbesartan: 3.3+/-0.1-fold induction; telmisartan: 3.1+/-0.3-fold induction; both P<0.01). Telmisartan showed a more pronounced induction of aP2 expression in lower, pharmacologically relevant concentrations compared with the other ARBs. The ARB losartan enhanced aP2 expression only at high concentrations (losartan 100 micromol/L: 3.6+/-0.3-fold induction; P<0.01), whereas eprosartan up to 100 micromol/L had no significant effects. In transcription reporter assays, irbesartan and telmisartan (10 micromol/L) markedly induced transcriptional activity of PPARgamma by 3.4+/-0.9-fold and 2.6+/-0.6-fold (P<0.05), respectively, compared with 5.2+/-1.1-fold stimulation by the PPARgamma ligand pioglitazone (10 micromol/L). Irbesartan and telmisartan also induced PPARgamma activity in an AT1R-deficient cell model (PC12W), demonstrating that these ARBs stimulate PPARgamma activity independent of their AT(1)R blocking actions. CONCLUSIONS: The present study demonstrates that a specific subset of ARBs induces PPARgamma activity, thereby promoting PPARgamma-dependent differentiation in adipocytes. The activation of PPARgamma demonstrates new pleiotropic actions of certain ARBs, providing a potential mechanism for their insulin sensitizing/antidiabetic effects. PMID- 15117842 TI - Phosphatidylinositol-3-kinase signaling is required for erythropoietin-mediated acute protection against myocardial ischemia/reperfusion injury. AB - BACKGROUND: Parenteral administration of recombinant human erythropoietin (rhEPO) to rats induces protection against myocardial ischemia/reperfusion injury 24 hours later. However, the mechanisms by which rhEPO mediates protection have not been determined. METHODS AND RESULTS: rhEPO was perfused into isolated rat hearts over 15 minutes immediately before 30 minutes of no-flow ischemia and 45 minutes of reperfusion. Compared with saline-perfused control hearts, recovery of left ventricular developed pressure was increased in rhEPO-perfused hearts. rhEPO also increased AKT activity and decreased apoptosis. All of these effects were blocked when the phosphatidylinositol-3-kinase inhibitor wortmannin was infused with rhEPO. CONCLUSIONS: rhEPO provides immediate protection against ischemia/reperfusion injury in the isolated perfused rat heart that is mediated by the phosphatidylinositol-3-kinase pathway. PMID- 15117843 TI - Thrombocytopenia caused by abciximab or tirofiban and its association with clinical outcome in patients undergoing coronary stenting. AB - BACKGROUND: Thrombocytopenia is a possible complication of treatment with glycoprotein (GP) IIb/IIIa antagonists during percutaneous coronary interventions, but it is not clear whether different GP IIb/IIIa inhibitors carry a different risk of thrombocytopenia, and its relation to clinical outcome is unknown. METHODS AND RESULTS: We analyzed data from the Do Tirofiban and Reopro Give Similar Efficacy Outcomes (TARGET) study, which compared the safety and efficacy of abciximab and tirofiban in patients undergoing coronary stenting. Platelets were measured at baseline and 6 and 24 hours after the beginning of treatment. Thrombocytopenia (nadir platelet count <100x10(9) cells/L) developed in 2.4% of patients treated with abciximab and 0.5% of those treated with tirofiban (P<0.001). The variables independently associated with thrombocytopenia were treatment with abciximab within the previous 6 months (OR, 4.4; 95% CI, 1.7 to 11.2), baseline creatinine levels of > or =0.8 mg/dL (OR, 3.8; 95% CI, 1.7 to 8.8), previous transient ischemic attack (OR, 3.2; 95% CI, 1.4 to 7.6), female gender (OR, 1.9; 95% CI, 1.2 to 3.1), and history of peripheral vascular disease (OR, 1.78; 95% CI, 1.0 to 3.1). Severe bleeding occurred more frequently in patients with thrombocytopenia (5.1% versus 0.7%, P=0.001), who also more frequently received blood transfusions (6.1% versus 1.4%, P=0.001). At the 30-day follow-up, 2.0% of patients with thrombocytopenia and 0.4% of those without (P=0.022) had died; myocardial infarction occurred in 9.13% versus 6.11% (P=NS); and target vessel revascularization occurred in 6.07% versus 0.60% (P<0.001). CONCLUSIONS: During coronary stenting, abciximab and other risk factors are independently associated with thrombocytopenia. Regardless of the cause, thrombocytopenia is associated with more ischemic events, bleedings, and transfusions. PMID- 15117844 TI - Detection of acutely impaired microvascular reperfusion after infarct angioplasty with magnetic resonance imaging. AB - BACKGROUND: Despite the reopening of the infarct-related artery (IRA) with infarct angioplasty, complete microvascular reperfusion does not always ensue. METHODS AND RESULTS: We performed cardiovascular MRI (CMR) in 20 acute myocardial infarction (AMI) patients within 24 hours of successful infarct angioplasty and 10 control patients without obstructive coronary artery disease on a clinical 1.5 T CMR scanner. Three-month follow-up CMR in AMI patients evaluated the impact of abnormal reperfusion on recovery of function. Infarction was localized by delayed contrast hyperenhancement and impaired systolic thickening. Microvascular perfusion was assessed at rest by first-pass perfusion CMR after a bolus of gadolinium-DTPA by use of the time to 50% maximum myocardial enhancement. Whereas contrast wash-in was homogeneous in control patients, AMI patients exhibited delays in the hypokinetic region subtended by the IRA compared with remote segments in 19 of 20 patients, with a mean contrast delay of 0.9+/-0.1 seconds (95% CI, 0.6 to 1.2 seconds). At follow-up, the mean recovery of systolic thickening was lower in segments with a contrast delay of 2 seconds or more (10+/ 7% versus 39+/-4%, P<0.001). A contrast delay > or =2 seconds and infarction >75% transmurally were independent predictors of impaired left ventricular systolic thickening at 3 months (P=0.002 for severe contrast delay, P=0.048 for >75% for transmural infarction). CONCLUSIONS: CMR detects impaired microvascular reperfusion in AMI patients despite successful infarct angioplasty, which when severe is associated with a lack of recovery of wall motion. PMID- 15117845 TI - Attenuation of oxidative stress and remodeling by cardiac inhibitor of metalloproteinase protein transfer. AB - BACKGROUND: Matrix metalloproteinase (MMP) and cardiac inhibitor of metalloproteinase (CIMP) are coexpressed in the heart. Although it is known that oxidative stress activates MMP and CIMP inhibits MMP, it is unclear whether CIMP administration attenuates oxidative stress and MMP-mediated cardiac dilatation. METHODS AND RESULTS: Arteriovenous fistula (AVF) was created in C57BL/J6 mice, and CIMP was administered to AVF and sham mice by protein transfer into peritoneal cavity by minipump for 4 weeks. Mice were grouped as follows: sham; sham+CIMP; AVF; and AVF+CIMP (n=6). In vivo left ventricular (LV) pressure was measured. Plasma and LV tissue levels of CIMP were measured by Western analysis. LV levels of NADPH oxidase activity, marker of oxidative stress, were increased in AVF mice and decreased in AVF mice treated with CIMP. Compared with sham, CIMP was decreased in AVF mice, and CIMP protein transfer increased plasma and LV tissue levels of CIMP in AVF mice; there was no increase in sham animals. In situ zymography demonstrated a robust increase in MMP activity in the hearts from AVF mice compared with sham, and treatment with CIMP decreased MMP activity. In AVF mice, the cardiac pressure-length relationship was similar to that observed in sham mice after administration of CIMP. Contractile responses of normal LV rings were measured in the presence and absence of CIMP. CIMP shifted the pressure length relationship to the left, attenuated LV dilatation, and had no effect on CaCl2-mediated contraction. CONCLUSIONS: Treatment of AVF mice with CIMP significantly abrogated the contractile dysfunction and decreased the oxidative stress in volume overload-induced heart failure. PMID- 15117846 TI - Propensity analysis of long-term survival after surgical or percutaneous revascularization in patients with multivessel coronary artery disease and high risk features. AB - BACKGROUND: Although most randomized clinical trials have suggested that long term survival rates after percutaneous coronary intervention (PCI) or surgical multivessel coronary revascularization (CABG) are equivalent, some post hoc analyses in high-risk groups and adjustment for severity of coronary disease have suggested higher mortality after PCI. METHODS AND RESULTS: We studied 6033 consecutive patients who underwent revascularization in the late 1990s. PCI was performed in 872 patients; 5161 underwent CABG. Half the patients had significant left ventricular dysfunction or diabetes. Propensity analysis to predict the probability of undergoing PCI according to 22 variables and their interactions was used. The C-statistic for this model was 0.90, indicating excellent discrimination between treatments. There were 931 deaths during 5 years of follow up. The 1- and 5-year unadjusted mortality rates were 5% and 16% for PCI and 4% and 14% for CABG (unadjusted hazard ratio, 1.13; 95% CI, 1.0 to 1.4; P=0.07). PCI was associated with an increased risk of death (propensity-adjusted hazard ratio, 2.3; 95% CI, 1.9 to 2.9; P<0.0001). This difference was observed across all categories of propensity for PCI and in patients with diabetes or left ventricular dysfunction. Other independent predictors of mortality (P< or =0.01 for all) were renal dysfunction, age, diabetes mellitus, chronic lung disease, peripheral vascular disease, left main trunk stenosis, and extent of coronary disease (Duke angiographic score). CONCLUSIONS: In patients with multivessel coronary artery disease and many high-risk characteristics, CABG was associated with better survival than PCI after adjustment for risk profile. PMID- 15117847 TI - Natriuretic peptides predict symptom-free survival and postoperative outcome in severe aortic stenosis. AB - BACKGROUND: The prognostic value of natriuretic peptides in aortic stenosis (AS) remains unknown. METHODS AND RESULTS: B-type natriuretic peptide (BNP), N terminal BNP (NtBNP), and N-terminal atrial natriuretic peptide (NtANP) were determined in 130 patients with severe AS (mean age, 70+/-12 years; mean gradient, 64+/-21 mm Hg; valve area, 0.64+/-0.15 cm2) who were followed up for 377+/-150 days. Natriuretic peptides increased with NYHA class and with decreasing ejection fraction (EF). Even asymptomatic patients frequently had elevated neurohormones. Asymptomatic patients who developed symptoms during follow-up had higher BNP and NtBNP levels at entry compared with those remaining asymptomatic (median for NtBNP, 131 pmol/L [interquartile range, 50 to 202 pmol/L] versus 31 pmol/L [range, 19 to 56 pmol/L]; P<0.001). Symptom-free survival at 3, 6, 9, and 12 months for patients with NtBNP <80 versus > or =80 pmol/L was 100%, 88+/-7%, 88+/-7%, and 69+/-13% compared with 92+/-8%, 58+/-14%, 35+/-15%, and 18+/-15%, respectively (P<0.001). Seventy-nine patients eventually underwent surgery because of symptoms. Considering preoperative neurohormone levels, age, NYHA class, aortic valve area, EF, and presence of coronary artery disease, we found that neurohormones, EF, and NYHA class predicted survival; neurohormones predicted postoperative symptomatic status; and neurohormones and preoperative EF predicted postoperative EF. However, by multivariate analysis, NtBNP was the only independent predictor of outcome. CONCLUSIONS: In severe AS, natriuretic peptides provide important prognostic information beyond clinical and echocardiographic evaluation. NtBNP independently predicts symptom-free survival, and preoperative NtBNP independently predicts postoperative outcome with regard to survival, symptomatic status, and left ventricular function. Thus, neurohormones may gain particular importance for timing of surgery in asymptomatic severe AS. PMID- 15117848 TI - Regionalization of treatment for subarachnoid hemorrhage: a cost-utility analysis. AB - BACKGROUND: Previous studies have shown that for the treatment of subarachnoid hemorrhage (SAH), outcomes are improved but costs are higher at hospitals with a high volume of admissions for SAH. Whether regionalization of care for SAH is cost-effective is unknown. METHODS AND RESULTS: In a cost-utility analysis, health outcomes for patients with SAH were modeled for 2 scenarios: 1 representing the current practice in California in which most patients with SAH are treated at the closest hospital and 1 representing the regionalization of care in which patients at hospitals with <20 SAH admissions annually (low volume) would be transferred to hospitals with > or =20 SAH admissions annually (high volume). Using a Markov model, we compared net quality-adjusted life-years (QALYs) and cost per QALY. Inputs were chosen from the literature and derived from a cohort study in California. Transferring a patient with SAH from a low- to a high-volume hospital would result in a gain of 1.60 QALYs at a cost of 10,548 dollars/QALY. For transfer to result in only borderline cost-effectiveness (50,000 dollars/QALY), differences in case fatality rates between low- and high volume hospitals would have to be one fifth as large (2.2%) or risk of death during transfer would have to be 5 times greater (9.8%) than estimated in the base case. CONCLUSIONS: Transfer of patients with SAH from low- to high-volume hospitals appears to be cost-effective, and regionalization of care may be justified. However, current estimates of the impact of hospital volume on outcome require confirmation in more detailed cohort studies. PMID- 15117849 TI - Osteoprotegerin is a risk factor for progressive atherosclerosis and cardiovascular disease. AB - BACKGROUND: Osteoprotegerin is a novel member of the tumor necrosis factor receptor superfamily and a soluble decoy receptor of the receptor activator of nuclear factor-kappaB ligand. Recent experimental research has implicated osteoprotegerin in atherogenesis, but epidemiological confirmation of this concept is sparse. METHODS AND RESULTS: As part of the prospective, population based Bruneck Study, severity, initiation, and progression of atherosclerosis were assessed in carotid arteries. Cases of incident cardiovascular disease and vascular mortality were carefully recorded over a 10-year period (1990 to 2000). Osteoprotegerin levels were measured in samples obtained at baseline and during follow-up. Serum osteoprotegerin showed a strong association with numerous vascular risk factors, including age, diabetes, markers of systemic inflammation, chronic infection, and smoking. In multivariate analyses, osteoprotegerin was significantly related to severity and 10-year progression of carotid atherosclerosis. Furthermore, a high level of osteoprotegerin was an independent risk factor for incident cardiovascular disease (adjusted relative risk for the top versus bottom tertile group for osteoprotegerin 2.2 [1.3 to 3.8]; P=0.001) and vascular mortality (adjusted relative risk for the top versus bottom tertile group for osteoprotegerin 3.1 [1.2 to 8.2]; P=0.010) but not for mortality due to nonvascular causes. CONCLUSIONS: Osteoprotegerin is an independent risk factor for the progression of atherosclerosis and onset of cardiovascular disease. PMID- 15117850 TI - Serum antibodies against Chlamydia pneumoniae outer membrane protein cross-react with the heavy chain of immunoglobulin in the wall of abdominal aortic aneurysms. AB - BACKGROUND: Chlamydia pneumoniae (Cp) has been demonstrated in arteries and abdominal aortic aneurysms (AAAs). However, the validity of the methods used is questioned, and antibiotic treatment trials have thus far shown disappointing results. Nevertheless, antibodies against the Cp outer membrane proteins (OMPs) have been associated with progression of atherosclerosis and AAAs. The aim of this study was to detect Cp OMPs in the wall of AAA patients by use of purified serum antibodies directed against Cp OMP and to assess potential cross-reacting proteins in AAA walls. METHODS AND RESULTS: Seventeen patients undergoing infrarenal AAA repair were studied. Full AAA thickness tissue was collected from the anterior wall of the aneurysm. Anti-OMP was extracted from seropositive AAA patients by use of an ELISA kit (Labsystems). Analysis was performed by use of 2D polyacrylamide gel electrophoresis, immunoblotting, and mass spectrometric protein identification. OMP antigens were not detected in 16 of 17 AAA walls. However, 3 major AAA proteins cross-reacted with anti-OMP. The proteins were all identified as heavy chains of human immunoglobulin. CONCLUSIONS: We could not find evidence of Cp OMP in 16 of 17 AAA walls, but instead, all samples showed a strong cross-reaction between Cp OMP antibodies and human immunoglobulin. This might indicate that AAA is an autoimmune disease, perhaps triggered by an initial Cp infection. PMID- 15117851 TI - Regulation of proangiogenic factor CCN1 in cardiac muscle: impact of ischemia, pressure overload, and neurohumoral activation. AB - BACKGROUND: CCN1, a potent proangiogenic factor, is induced in the vasculature by tissue injury, angiotensin II (Ang II), and growth factor stimulation. Because these conditions occur in myocardial ischemia and pressure overload, we investigated the regulation of CCN1 in cardiomyocytes in vitro and in the heart in vivo. METHODS AND RESULTS: Ang II, signaling via the angiotensin type 1 (AT1) receptor, and alpha1-adrenergic stimulation with phenylephrine induced CCN1 expression in ventricular cardiomyocytes isolated from 1- to 3-day-old rats. Cell culture supernatant of Ang II-treated cardiomyocytes induced migration of smooth muscle cells, which was abolished by neutralizing antibody to CCN1. Ang II- and phenylephrine-mediated induction of CCN1 expression in cardiomyocytes was completely abolished by inhibition of MEK/extracellular signal-regulated kinases (ERK) or protein kinase C (PKC). Likewise, mechanical stretch induced CCN1 expression in cardiomyocytes, an effect that was prevented by AT1 receptor blockade or PKC inhibition. Similarly, pressure overload in vivo upregulated myocardial CCN1 expression levels via AT1 receptor- and PKC-dependent mechanisms. After myocardial infarction in mice, CCN1 expression was strongly induced in both ischemic and remote left ventricular myocardium. Marked CCN1 protein expression was noted in cardiomyocytes of patients with end-stage ischemic cardiomyopathy but was almost absent in nonfailing human myocardium. CONCLUSIONS: Pressure overload, ischemia, and neurohormonal factors, such as Ang II or alpha1 adrenergic stimuli, induce myocardial expression of CCN1, a potent proangiogenic factor, supporting the notion that CCN1 may play an important role in the adaptation of the heart to cardiovascular stress. PMID- 15117852 TI - Sympathetic drive in anterior and inferior uncomplicated acute myocardial infarction. AB - BACKGROUND: The sympathetic activation that follows acute myocardial infarction (AMI) has been associated with increased morbidity and mortality. Because the prognosis after anterior AMI (ant-AMI) is worse than that after inferior AMI (inf AMI), we planned to determine whether the magnitude of sympathetic hyperactivity differs between the two. METHODS AND RESULTS: Thirty-nine patients with uncomplicated AMI, comprising 2 matched groups of 17 patients with ant-AMI, and 22 patients with inf-AMI were examined. Measurements were obtained 2 to 4 days after AMI and compared with 20 normal subjects (NC) who were matched in terms of age and body weight to the AMI groups. Resting muscle sympathetic nerve activity was quantified from multiunit bursts (MSNA) and from single units (s-MSNA). Both groups of AMI patients were matched with regard to hemodynamic variables, left ventricular function, and infarct size. Both groups had greater (at least P<0.01) sympathetic nerve activity than NC (60+/-4.3 bursts/100 cardiac beats and 68+/ 4.9 impulses/100 cardiac beats), but the magnitude of sympathetic nerve hyperactivity in ant-AMI (81+/-4.0 bursts/100 cardiac beats and 91+/-4.9 impulses/100 cardiac beats) was similar (P>0.05) to that in inf-AMI (80+/-3.2 bursts/100 cardiac beats and 90+/-4.0 impulses/100 cardiac beats) CONCLUSIONS: Both ant-AMI and inf-AMI resulted primarily in a similar magnitude of sympathetic nerve hyperactivity. These findings suggest that the worse prognosis after ant AMI compared with after inf-AMI would not be related primarily to the degree of sympathetic hyperactivity. PMID- 15117853 TI - Serum levels of the interleukin-1 receptor family member ST2 predict mortality and clinical outcome in acute myocardial infarction. AB - BACKGROUND: Mechanically overloaded cardiomyocytes secrete a soluble interleukin 1 receptor family member called ST2. Serum levels of ST2 are associated with prognosis in nonischemic heart failure, but the predictive value of ST2 in patients with acute myocardial infarction is unknown. METHODS AND RESULTS: ST2 levels were measured in serum from 810 patients with acute myocardial infarction in the Thrombolysis In Myocardial Infarction (TIMI) 14 (362 patients) and Enoxaparin and TNK-tPA With or Without GPIIb/IIIa Inhibitor as Reperfusion Strategy in STEMI (ENTIRE)-TIMI 23 (448 patients) clinical trials. Baseline levels of ST2 were significantly higher in those patients who died (0.379 versus 0.233 ng/mL, P=0.0001) or developed new congestive heart failure (0.287 versus 0.233 ng/mL, P=0.009) by 30 days. In an analysis of outcomes at 30 days by ST2 quartiles, both death (P=0.001) and the combined death/heart failure end point (P=0.001) showed a significant graded association with levels of ST2; furthermore, in-hospital death (P=0.003) and death/heart failure (P=0.004) were also significantly associated with higher ST2 levels. In a logistic regression analysis that controlled for important clinical factors, increasing levels of ST2 remained associated with death at 30 days (P=0.047). ST2 levels rose during the first day after infarction and were maximal at 12 hours; ST2 levels at 12 hours were also independently associated with death at 30 days (P<0.001). CONCLUSIONS: Serum levels of the interleukin-1 receptor family member ST2 predict mortality and heart failure in patients with acute myocardial infarction. These data suggest that ST2 may be a useful biomarker and that this novel inflammatory receptor may play a role in cardiac pathophysiology. PMID- 15117854 TI - Endothelium-derived hyperpolarizing factor-mediated renal vasodilatory response is impaired during acute and chronic hyperhomocysteinemia. AB - BACKGROUND: Endothelial dysfunction is an early event in the development of vascular complications in hyperhomocysteinemia. Endothelial cells release a number of vasodilators, including NO and prostacyclin. Several lines of evidence have indicated the existence of a third vasodilator pathway, mediated by endothelium-derived hyperpolarizing factor (EDHF). EDHF is a major determinant of vascular tone in small resistance vessels. The influence of hyperhomocysteinemia on EDHF is unknown. The present in vivo study evaluates the integrity of the EDHF pathway in the renal microcirculation of rats with acute and chronic hyperhomocysteinemia. METHODS AND RESULTS: EDHF-mediated vasodilation was evaluated as the renal blood flow (RBF) response to intrarenal acetylcholine during systemic NO synthase and cyclooxygenase inhibition. Acute hyperhomocysteinemia induced by intravenous homocysteine did not affect EDHF mediated vasodilation. In contrast, intravenous methionine with subsequent hyperhomocysteinemia impaired the EDHF-mediated RBF response. When the methionine infusion was preceded by adenosine periodate oxidized to prevent the cleavage of S-adenosylhomocysteine to homocysteine and adenosine, a similar impairment of EDHF was observed, but with normal homocysteine levels. Animals with chronic hyperhomocysteinemia induced by a high-methionine, low-B vitamin diet during 8 weeks had a severely depressed EDHF-mediated vasodilation compared with those on a standard diet. Endothelium-independent vasodilation to deta-NONOate and pinacidil was not affected in acute and chronic hyperhomocysteinemia, demonstrating intact vascular smooth muscle reactivity. CONCLUSIONS: EDHF dependent responses are impaired in the kidney of hyperhomocysteinemic rats. Because EDHF is a major regulator of vascular function in small vessels, these findings have important implications for the development of microangiopathy in hyperhomocysteinemia. PMID- 15117855 TI - Rapid intraoperative immunoassay of parathyroid hormone and other hormones: a new paradigm for point-of-care testing. AB - BACKGROUND: The first description of the use of a rapid assay for the measurement of intact parathyroid hormone (PTH) in patients undergoing parathyroidectomy for hyperparathyroidism was reported in 1988. Subsequent improvements in the analytical performance of the rapid intraoperative PTH assay allowed the establishment of its clinical utility in the surgical management of hyperparathyroidism. These modifications also allowed the assay to be performed in or near the operating suite. METHODS: We searched MEDLINE, using the following key words: intraoperative, rapid, quick, parathyroid hormone, hormone, and immunoassay. Relevant articles that focused on the analytical aspects and clinical utility of rapid intraoperative hormone immunoassays were selected for this review. CONTENT: On the basis of the positive impact that the rapid intraoperative PTH test has had on both patient outcomes and cost savings, other rapid intraoperative hormone immunoassays for the diagnosis and/or treatment of other endocrine-hormone-secreting tumors have been developed. These hormones share certain characteristics that make them suitable for use as rapid intraoperative tests, i.e., short analyte half-life and/or large analyte concentration gradient, rapid analysis time, and positive clinical utility. Initial studies with cortisol, gastrin, insulin, adrenocorticotropic hormone, and testosterone have shown promising results in preoperative localization studies and/or for assessing the effectiveness of tumor resection during surgery. CONCLUSION: The emergence of these rapid intraoperative immunoassays indicates that this test format is likely to provide future opportunities to improve patient care by advances in clinical laboratory testing. PMID- 15117856 TI - Simultaneous determination of alpha-fetoprotein and free beta-human chorionic gonadotropin by element-tagged immunoassay with detection by inductively coupled plasma mass spectrometry. AB - BACKGROUND: An inductively coupled plasma mass spectrometry (ICP-MS)-based immunoassay has been proposed independently by Baranov et al. (Anal Chem 2002;74:1629-36) and our group, but the applicability of this method for multianalyte analysis in clinical samples has not been fully illustrated. We developed a dual-label immunoassay method for the simultaneous determination of alpha-fetoprotein (AFP) and free beta-human chorionic gonadotropin (hCGbeta) in human serum. METHODS: Monoclonal antibodies immobilized on microtiter plates captured AFP and hCGbeta, which were detected by use of Eu(3+)-labeled anti-AFP and Sm(3+)-labeled anti-hCGbeta monoclonal antibodies. Eu(3+) and Sm(3+) were dissociated from the immunocomplex with HNO(3) solution (10 mL/L) and delivered by peristaltic pump to the ICP mass spectrometer. RESULTS: The measurable ranges of AFP and hCGbeta were 4.6-500 and 5.0-170 microg/L, respectively, with detection limits of 1.2 and 1.7 microg/L (3 SD above mean of zero calibrator), respectively. The intraassay imprecision (CV) for AFP was 8.3%, 4.0%, and 2.7% at 16.3, 86, and 354 microg/L, respectively, and the interassay CV was 10%, 5.7%, and 3.5%. For hCGbeta, the intraassay CV was 5.4%, 6.4%, and 3.1%, respectively, at 10.5, 45.2, and 105 microg/L, and the interassay CV was 7.2%, 8.0%, and 3.7%. Comparison with IRMAs for AFP and hCGbeta yielded correlation coefficients (r(2)) of 0.97 and 0.95. CONCLUSIONS: Two proteins can be measured simultaneously by immunoassays using two rare earth elemental tags (Eu(3+) and Sm(3+)) and ICP-MS detection. The multielement capability and the multiple potential elemental labels make ICP-MS attractive for multianalyte immunoassays. Implementation of ICP-MS-linked immunoassays may be relatively straightforward because the labeling and immunoreaction procedures have been well developed for clinical time-resolved immunofluorometric assays. PMID- 15117857 TI - Pioglitazone reduces atherogenic index of plasma in patients with type 2 diabetes. AB - BACKGROUND: Insulin resistance is often associated with increased triglyceride (TG) and decreased HDL-cholesterol (HDL-C) concentrations and increased small LDL particles. The Atherogenic Index of Plasma (AIP), defined as log(TG/HDL-C), has recently been proposed as a marker of plasma atherogenicity because it is increased in people at higher risk for coronary heart disease and is inversely correlated with LDL particle size. We studied the effect of pioglitazone, a thiazolidinedione that reduces insulin resistance, on the AIP of patients with type 2 diabetes. METHODS: The data for the analysis of AIP in this report were obtained from four randomized, double-blind, multicenter, parallel-group, placebo controlled clinical trials. Pioglitazone was used as monotherapy in one study and in combination therapy in three studies. Fasting glucose, insulin, HDL-C, and TGs plus glycohemoglobin (HbA(1C)) were measured at baseline and various points during each study. RESULTS: Patients in this study population with type 2 diabetes had high AIP values at baseline. Pioglitazone treatment significantly decreased AIP from baseline in each of the study groups. Pioglitazone treatment groups had a significantly lower AIP compared with their respective placebo controls. Finally, AIP was inversely and significantly correlated with measures of insulin sensitivity, such as the homeostasis model assessment and quantitative insulin sensitivity check index. In contrast, AIP was not significantly correlated with HbA(1C). CONCLUSIONS: Pioglitazone reduced AIP when used as monotherapy or in combination therapy with sulfonylurea, metformin, or insulin. AIP was inversely correlated with measures of insulin sensitivity. PMID- 15117858 TI - A chicken in every pot and a drug-eluting stent in every lesion. PMID- 15117859 TI - The childhood obesity epidemic: impact on endothelial function. PMID- 15117860 TI - C-reactive protein: structure affects function. PMID- 15117861 TI - Elevated remnant-like particle cholesterol concentration: a characteristic feature of the atherogenic lipoprotein phenotype. PMID- 15117862 TI - Images in cardiovascular medicine. Univentricular heart. PMID- 15117863 TI - Physical activity and exercise recommendations for stroke survivors: an American Heart Association scientific statement from the Council on Clinical Cardiology, Subcommittee on Exercise, Cardiac Rehabilitation, and Prevention; the Council on Cardiovascular Nursing; the Council on Nutrition, Physical Activity, and Metabolism; and the Stroke Council. PMID- 15117864 TI - Images in cardiovascular medicine. Proximal subclavian artery stenosis diagnosis and repair documented by both myocardial perfusion imaging and angiography. PMID- 15117865 TI - Serum myoglobin in pulmonary embolism. PMID- 15117866 TI - Why did the anti-C5 complement antibody pexelizumab not reduce infarct size but influence clinical outcomes positively when applied as adjunctive therapy to primary percutaneous coronary intervention? PMID- 15117867 TI - Sildenafil use in patients with the Eisenmenger syndrome. PMID- 15117868 TI - Unique female indicators of coronary heart disease: do they exist? PMID- 15117869 TI - Hyperinsulinemia, hyperleptinemia, and nitric oxide in the regulation of membrane micoviscosity . PMID- 15117870 TI - Left ventricular noncompaction suggests myopathy. PMID- 15117872 TI - Developmental regulation of gonadotropin-releasing hormone neurons by fibroblast growth factor signaling. AB - GnRH neurons are central to the initiation and maintenance of reproductive function in diverse vertebrates. The formation of a functional GnRH system during development is a highly complex event that likely requires extensive guidance by neurotrophic factors. In this study, we examined whether members of the fibroblast growth factor (FGF) family are critically involved in the development of endogenous GnRH neurons. Immunocytochemistry revealed the presence of FGF receptors (FGFRs) 1, 2, and 3, but not 4, in embryonic day (E) 10.5 medial nasal placode, an area and time consistent with the first appearance of GnRH neurons in mice. Dual immunocytochemistry confirmed the presence of FGFRs 1 and 3, but not 2 and 4, in a substantial fraction of E15.5 and postnatal day (P) 3 GnRH neurons. To examine whether FGF signaling was essential for the specification of GnRH neuronal fate, a nasal explant culture that supported the in vitro emergence of GnRH neurons from E10.5 noses was established. In this system, the addition of SU5402, a FGFR antagonist, suppressed the emergence of GnRH neurons. Lastly, we investigated whether FGF signaling altered the extension of neurites in cultures of dispersed GnRH neurons. The addition of FGF2 to E15.5 and P3 GnRH neurons expressing the green fluorescent protein significantly stimulated neurite outgrowth (E15.5 and P3) and branching (P3), suggesting a regulatory role of FGFs in GnRH axon targeting. Together these results demonstrated that FGF signaling critically regulates multiple phases of development in a neuroendocrine system essential for vertebrate reproduction. PMID- 15117873 TI - Metabolic effects of transgenic melanocyte-stimulating hormone overexpression in lean and obese mice. AB - The proopiomelanocortin-derived peptide, alpha-MSH, inhibits feeding via melanocortin receptors in the hypothalamus and genetic defects inactivating the melanocortin system have been shown to lead to obesity in experimental animals and humans. To determine whether long-term melanocortinergic activation has significant effects on body weight and composition and insulin sensitivity, transgenic mice overexpressing N-terminal proopiomelanocortin, including alpha- and gamma(3)-MSH, under the control of the cytomegalovirus-promoter were generated. The transgene was expressed in multiple tissues including the hypothalamus, in which both alpha-MSH and gamma(3)-MSH levels were increased approximately 2-fold, compared with wild-type controls. Transgene homozygous mice were also crossed with obese leptin receptor-deficient db(3J) and obese yellow A(y) mice. MSH overexpression led to uniform, dose- dependent darkening of coat color. MSH overexpression reduced weight gain and adiposity and improved glucose tolerance in lean male mice. In female transgenic mice, there was no significant effect on body weight, but there was a significant decrease in insulin levels. Obesity was attenuated in obese db(3J)/db(3J) male and female mice, but there was no improvement in glucose metabolism. In contrast, the MSH transgene improved glucose tolerance in male A(y) mice. These results support the hypothesis that long-term melanocortinergic activation could serve as a potential strategy for anti-obesity and/or antidiabetic therapy. PMID- 15117874 TI - Fate of internalized thyrotropin-releasing hormone receptors monitored with a timer fusion protein. AB - Trafficking of TRH receptors was studied in a stable HEK293 cell line expressing receptor fused to a Timer protein (TRHR-Timer) that spontaneously changes from green to red over 10 h. Cells expressing TRHR-Timer responded to TRH with an 11 fold increase in inositol phosphate formation, increased intracellular free calcium, and internalization of 75% of bound [(3)H][N(3)-methyl-His(2)]TRH within 10 min. After a 20-min exposure to TRH at 37 C, 75-80% of surface binding sites disappeared as receptors internalized. When TRH was removed and cells incubated in hormone-free medium, approximately 75% of [(3)H][N(3)-methyl-His(2)]TRH binding sites reappeared at the surface over the next 2 h with or without cycloheximide. Trafficking of TRHR-Timer was monitored microscopically after addition and withdrawal of TRH. In untreated cells, both new (green) and old (red) receptors were seen at the plasma membrane, and TRH caused rapid movement of young and old receptors into cytoplasmic vesicles. When TRH was withdrawn, some TRHR-Timer reappeared at the plasma membrane after several hours, but much of the internalized receptor remained intracellular in vesicles that condensed to larger structures in perinuclear regions deeper within the cell. Strikingly, receptors that moved to the plasma membrane were generally younger (more green) than those that underwent endocytosis. There was no change in the red to green ratio over the course of the experiment in cells exposed to vehicle. The results indicate that, after agonist-driven receptor internalization, the plasma membrane is replenished with younger receptors, arising either from an intracellular pool or preferential recycling of younger receptors. PMID- 15117875 TI - Signal transducer and activator of transcription (Stat)-6-dependent, but not Stat4-dependent, immunity is required for the development of autoimmunity in Graves' hyperthyroidism. AB - The role of T helper (Th) cells in experimental models of Graves' hyperthyroidism is still somewhat controversial. To further investigate the role of Th1- and Th2 dependent immunity during the development of Graves' hyperthyroidism, we tested mice with targeted deletion of signal transducer and activator of transcription-4 (Stat4) or Stat6 genes that, respectively, have impaired Th1 and Th2 immunity. We immunized wild-type BALB/c, Stat4(-/-), or Stat6(-/-) mice with human embryonic kidney cells (293 cells) expressing the extracellular domain of human TSH receptor (293-TBP cells). Fifty percent of wild-type BALB/c and Stat4(-/-) mice developed Graves' hyperthyroidism with elevated serum T(4) levels and thyroid stimulatory antibodies. In contrast, Stat6(-/-) mice resisted development of the disease. Stat4(-/-) mice exhibited a dominant Th2 immune response characterized by the production of IL-4 and IgG1 anti-TSH receptor antibodies. However, Stat6( /-) mice displayed a strong Th1 immune response characterized by the production of interferon-gamma and IgG2a antibodies. Hyperthyroid mice showed enlargement of thyroid glands with hypertrophy and decreased amounts of colloid material, all characteristics of Graves' disease. These data demonstrate that in this model, Stat6-dependent Th2 immunity is critical for the development of Graves' hyperthyroidism. PMID- 15117876 TI - Liver receptor homolog-1 stimulates the progesterone biosynthetic pathway during follicle-stimulating hormone-induced granulosa cell differentiation. AB - FSH-stimulated granulosa cell differentiation is associated with the induction of the LH receptor (LHr) as well as induction of the estrogen and progesterone biosynthetic pathways. Although activation of the cAMP-protein kinase A pathway is sufficient to stimulate progesterone production, additional pathways are required for the induction of the LHr and p450 aromatase. The orphan nuclear receptor, liver receptor homolog-1 (LRH-1), is expressed in granulosa cells and has been shown to synergize with the cAMP signaling system to regulate the gonadal type II aromatase promoter in transient transfection assays. To determine whether LRH-1 can interact with the cAMP pathway in the induction of aromatase and the LHr, we examined the effects of an adenoviral vector that directs the expression of human LRH-1 (Ad-LRH-1) on FSH-stimulated granulosa cell differentiation. Infection of undifferentiated granulosa cells with LRH-1 alone had no effect on estrogen production, progesterone production, or the expression of the LHr. However, combination of FSH stimulation and Ad-LRH-1 infection led to significantly greater progesterone production and increases in mRNA for p450 side chain cleavage and 3beta-hydroxysteroid dehydrogenase than granulosa cells stimulated by FSH alone. However, infection with Ad-LRH-1 did not stimulate estradiol production or increases in mRNA for p450 aromatase or the LHr above that seen with FSH treatment alone. Moreover, infection with Ad-LRH-1 was able to overcome H-89 inhibition of FSH-stimulated progesterone but not estrogen production. Collectively, these observations support a direct role for LRH-1 in the induction of the progesterone but not the estrogen biosynthetic pathway during granulosa cell differentiation. PMID- 15117877 TI - Circulating triglycerides impact on orexigenic peptides and neuronal activity in hypothalamus. AB - Little is known about the impact of circulating lipids on brain processes. Building on evidence that chronic fat consumption stimulates hypothalamic peptides in close association with elevated triglycerides (TG), this study examined whether an acute rise in TG levels induced by fat emulsion can affect these hypothalamic systems. In normal weight rats, ip injection of Intralipid (20%, 5 ml) during the first 4 h after injection produced a robust increase in TG levels and nonesterified fatty acids, but had no impact on glucose, insulin, or leptin levels. This was accompanied by a marked increase in the expression of particular orexigenic peptides, galanin, orexins, and the opioid, enkephalin, which are known to be positively related to fat ingestion. This effect, similarly induced by 4 h of high fat diet consumption, was detected in the paraventricular nucleus (PVN) for galanin, in the perifornical hypothalamus (PFH) for orexins, and in the PVN, PFH, as well as the arcuate nucleus (ARC) for enkephalin. It was not seen, however, for neuropeptide Y and agouti-related protein localized in the ARC, which are unaffected or reduced by dietary fat. This site specificity was confirmed by c-Fos immunostaining, a marker of neuronal activity, which was increased by Intralipid in the PVN and PFH, but not in the ARC, and was detected in 20% of orexin-expressing neurons in the PFH. These findings suggest that circulating lipids, through different mechanisms, may stimulate hypothalamic neurons, which synthesize specific feeding stimulatory peptides that possibly contribute to hyperphagia during consumption of a fat-rich diet. PMID- 15117878 TI - The basic residues in the membrane-proximal C-terminal tail of the rat melanin concentrating hormone receptor 1 are required for receptor function. AB - Melanin-concentrating hormone (MCH) is a hypothalamic neuropeptide that plays a key role in food intake. It acts through two G protein-coupled receptors (GPCRs), MCH1R and MCH2R, of which MCH1R is the primary regulator of food intake. We have previously reported that N-linked glycosylation of the extracellular domain of MCH1R is necessary for cell surface expression and signal transduction. We now report a role for the rat MCH1R C-terminal region. We constructed serial C terminal truncation mutants and determined the resulting changes in protein expression, cell surface expression, ligand binding, and MCH-stimulated calcium influx. By analyzing two mutants, deltaT317 (deletion of 36 C-terminal amino acids) and deltaR321 (deletion of 32 C-terminal amino acids), we found that the region between Phe(318) and Arg(321)) was responsible for signal transduction. A more detailed analysis was performed with single or multiple residue mutations. Single mutations of Arg(319), Lys(320), or Arg(321) exhibited a decrease in the cell surface expression, whereas mutations of either Arg(319) or Lys(320), but not Arg(321), showed a significant reduction in the calcium influx. Furthermore, simultaneous mutations of Arg(319) and Lys(320) produced a pronounced decrease in the efficacy of calcium influx stimulation compared with single mutations. A computational analysis revealed a dibasic amino acid motif that is conserved among many class 1 GPCRs and may be part of the amphiphilic cytoplasmic helix 8 (an eight-cytoplasmic helix). Our results therefore provide new insights into the role of the putative helix 8 in the regulation of GPCR function. PMID- 15117879 TI - The role of cysteines and charged amino acids in extracellular loops of the human Ca(2+) receptor in cell surface expression and receptor activation processes. AB - The Ca(2+) receptor is a plasma-membrane bound G protein-coupled receptor stimulated by extracellular calcium [Ca(2+)](o) and other di- and poly-cations. We investigated the role in receptor activation of all the charged amino acid residues and cysteines in the three extracellular loops (EL1, 2, and 3) of the human Ca(2+) receptor by alanine-scanning mutagenesis. The mutant receptors were transiently expressed in HEK-293 cells, and cell surface expression patterns were analyzed by endoglycosidase-H digestion, immunoblotting, intact cell ELISA, and hydrolysis of phosphoinositides (PI) induced by [Ca(2+)](o.) The mutation of Cys677 and Cys765 located in EL1 and EL2, respectively, ablated PI hydrolysis completely, showed less than 5% cell surface expression of the wild-type receptor, and were not properly glycosylated. Replacement of the charged residues by using a single mutation or multiple alanine mutations in EL1, 2, and 3 produced only minor changes in receptor activation, except for Glu767 and Lys831. The E767A and K831A mutations in EL2 and EL3, respectively, showed gain-of function by significantly enhancing apparent [Ca(2+)](o) affinity. E767A and K831A exhibited EC(50) values of 2.1 and 2.8 mm, respectively, for [Ca(2+)](o) stimulated PI hydrolysis as opposed to EC(50) value of 4.2 mm for the wild-type receptor. Like E767A, substitutions of Glu767 with Gln and Lys was similarly activating, whereas Asp substitution displayed wild-type [Ca(2+)](o) sensitivity. Substitution of Lys831 with Glu but not with Gln showed similar activating effect as Ala replacement. A double-mutant E767K/K831E in which charged residues were switched positions showed impaired cell surface expression and failed to respond to [Ca(2+)](o.) Taken together, these results suggest that in ELs, two cysteines form critical disulfide links, and the side chains of Glu767 and Lys831 are probably involved in ionic interactions with other prospective oppositely charged residues. Some of these interactions could be important for receptor folding and also may contribute to keep the Ca(2+) receptor transmembrane helix bundle in an inactive conformation. PMID- 15117880 TI - Crucial role of activin a in tubulogenesis of endothelial cells induced by vascular endothelial growth factor. AB - The present study was conducted to elucidate the role of activin A in tubulogenesis of vascular endothelial cells. Activin A was produced in bovine aortic endothelial cells (BAEC). These cells also expressed the type I and type II activin receptors. When added to BAEC cultured in a collagen gel, activin A induced capillary formation. Activin A was as potent as vascular endothelial growth factor (VEGF) and markedly enhanced VEGF-induced tubulogenesis. To examine the role of endogenous activin A, we added follistatin, an inhibitor of activin A. Follistatin nearly completely blocked the VEGF-induced tubulogenesis, and the effect of follistatin was reproduced by transfection of the dominant-negative type II activin receptor gene. In BAEC, activin A increased the expression of VEGF and the VEGF receptors, Flt-1 and Flk-1. On the other hand, VEGF increased the production of activin A. Finally, addition of follistatin, which blocks the action of endogenous activin A, reduced the expression of Flt-1 and Flk-1. These results indicate that an autocrine factor activin A amplifies the effect of VEGF by up-regulating VEGF and its receptors. This effect of activin A is critical in the VEGF-induced tubulogenic morphogenesis in BAEC. PMID- 15117881 TI - Aggregation and lack of secretion of most newly synthesized proinsulin in non beta-cell lines. AB - Myoblasts transfected with HB10D insulin secrete more hormone than those transfected with wild-type insulin, as published previously, indicating that production of wild-type insulin is not efficient in these cells. The ability of non-beta-cells to produce insulin was examined in several cell lines. In clones of neuroendocrine GH(4)C(1) cells stably transfected with proinsulin, two thirds of (35)S-proinsulin was degraded within 3 h of synthesis, whereas (35)S-prolactin was stable. In transiently transfected neuroendocrine AtT20 cells, half of (35)S proinsulin was degraded within 3 h after synthesis, whereas (35)S-GH was stable. In transiently transfected fibroblast COS cells, (35)S-proinsulin was stable for longer, but less than 10% was secreted 8 h after synthesis. Proinsulin formed a concentrated patch detected by immunofluorescence in transfected cells that did not colocalize with calreticulin or BiP, markers for the endoplasmic reticulum, but did colocalize with membrin, a marker for the cis-medial Golgi complex. Proinsulin formed a Lubrol-insoluble aggregate within 30 min after synthesis in non-beta-cells but not in INS-1E cells, a beta-cell line that normally produces insulin. More than 45% of (35)S-HB10D proinsulin was secreted from COS cells 3 h after synthesis, and this mutant formed less Lubrol-insoluble aggregate in the cells than did wild-type hormone. These results indicate that proinsulin production from these non-beta-cells is not efficient and that proinsulin aggregates in their secretory pathways. Factors in the environment of the secretory pathway of beta-cells may prevent aggregation of proinsulin to allow efficient production. PMID- 15117882 TI - Do human vascular endothelial cells produce aldosterone? AB - Extraadrenal production of aldosterone has been reported in several tissues, including vascular endothelial cells. The implications of local production of aldosterone in certain nonepithelial target tissues in normal and pathological physiology could be very important and merits further investigation. Human vascular endothelial cells have been reported to synthesize aldosterone under the regulation of angiotensin II. However, discrepancies are noted upon close scrutiny, the most important of which are the relative large efficiency of deoxycorticosterone conversion to aldosterone and the rate of aldosterone production in comparison to the adrenal zona glomerulosa cells. We investigated the production of aldosterone in three different human vascular endothelial cell lines, two from human umbilical veins, one from human pulmonary artery endothelial cells using a very sensitive ELISA method. Cells were incubated with the secretagogues angiotensin II, ACTH, and K(+), at various physiological concentrations with and without 1 microm deoxycorticosterone as additional substrate. In addition, RT-PCR was used to detect expression of the mRNA for the aldosterone synthase gene using a protocol developed by us that detects very low expression in subregions of the human brain. Our results failed to demonstrate mRNA for the aldosterone synthase gene or aldosterone biosynthesis in human endothelial cells. PMID- 15117883 TI - Is peptide bond cis/trans isomerization a key stage in the chemo-mechanical cycle of motor proteins? AB - Motor proteins such as myosin and kinesin are responsible for actively directed movement in vivo. The physicochemical mechanism underlying their function is still obscure. A novel and unifying model concerning the motors driving mechanism is suggested here. This model resides within the framework of the well-studied "swinging lever-arm" hypothesis, stating that cis/trans peptide bond isomerization (CTI) is a key stage in the chemo-mechanical coupling within actomyosin--the complex of the motor (myosin) and its specific track (actin). CTI is suggested to propel myosin's lever-arm swing. The model addresses on the submolecular level a broad spectrum of actomyosin's functional characteristics, such as kinetics, energetics, force exertion, stepping, and directionality. The model may be tested first with relative ease in kinesin--a smaller motor that could be specifically modified with unnatural amino acids using bacterial expression. Suggested modifications may be used for labeling and functional decoupling. PMID- 15117884 TI - Cyclooxygenases: new forms, new inhibitors, and lessons from the clinic. AB - The beneficial actions of nonsteroidal anti-inflammatory drugs (NSAIDs) have been linked to their ability to inhibit inducible COX-2 at sites of inflammation, and their side effects (e.g., gastric damage) to inhibition of constitutive COX-1. Selective inhibitors of COX-2, such as celecoxib, etoricoxib, lumiracoxib, rofecoxib, and valdecoxib have been developed and the greatest recent growth in our knowledge in this area has been come from the clinical use of these compounds. Although clinical data indicate that COX-2 selectivity is associated with a reduction in severe gastrointestinal events, they also reveal there are roles for constitutive COX-2 within tissues such as the brain, kidney, pancreas, intestine, and blood vessels. We now better understand the roles of COX-1 and COX 2 in functions as disparate as the perception of pain and the progression of cancers. Clinical use of COX-2-selective compounds has ignited strong debates regarding potential side effects, most notably those within the cardiovascular system such as myocardial infarctions, strokes, and elevation in blood pressure. This review will discuss how the latest studies help us understand the roles of COX-1 and COX-2 and what clinically proven benefits the newer generation of COX-2 selective inhibitors offer PMID- 15117885 TI - Modulatory effects of HMG-CoA reductase inhibitors in diabetic microangiopathy. AB - 3-hydroxy-3-methyl-glutaryl CoA (HMG-CoA) reductase inhibitors or statins are competitive inhibitors of the rate-limiting enzyme in cholesterol biosynthesis. Several large landmark clinical studies have shown a marked reduction of cardiovascular mortality and morbidity in patients treated with statins. Because of the strong association between serum cholesterol levels and coronary artery disease, investigators initially assumed that the predominant beneficial effects of statins result from their lipid-lowering properties. However, more recent observations have suggested that the clinical benefits of statins may be in part independent of their cholesterol-lowering effects. The pleiotropic or cholesterol independent effects of statins might result from preventing the production of isoprenoids. Isoprenoids serve as important lipid attachments for the post translational modification of a variety of proteins such as small GTP binding proteins implicated in intracellular signaling. The list of different pleiotropic effects of statins is still growing and, among others, includes the modulatory effects of statins on endothelial function, oxidative stress, coagulation, plaque stability, and inflammation. The pleiotropic effects of statins represent an area of great interest in prevention and therapy of cardiovascular and other chronic diseases. An area of particular interest is the potential beneficial effects of statins in diabetes and its micro/macrovascular complications. This review summarizes our current understanding of the pleiotropic effects of statins in diabetes and the modulatory effects of statins in various pathobiological pathways involved in diabetes and its complications. PMID- 15117886 TI - TGF-beta signaling and the fibrotic response. AB - The cause of fibrotic diseases, pathologies characterized by excessive production, deposition, and contraction of extracellular matrix, is unknown. To understand the molecular basis of fibrotic disease, it is essential to appreciate how matrix deposition is normally controlled and how this process is dysregulated in fibrogenesis. This review discusses the current state of knowledge concerning interactions among the profibrotic proteins transforming growth factor-beta (TGF beta), connective tissue growth factor (CTGF, CCN2), and ED-A fibronectin (ED-A FN) and the antifibrotic proteins tumor necrosis factor-alpha (TNF-alpha) and gamma-interferon (IFN-gamma). PMID- 15117887 TI - Prevention of cardiac fibrosis and left ventricular dysfunction in diabetic cardiomyopathy in rats by transgenic expression of the human tissue kallikrein gene. AB - Diabetic cardiomyopathy includes fibrosis. Kallikrein (KLK) can inhibit collagen synthesis and promote collagen breakdown. We investigated cardiac fibrosis and left ventricular (LV) function in transgenic rats (TGR) expressing the human kallikrein 1 (hKLK1) gene in streptozotocin (STZ) -induced diabetic conditions. Six weeks after STZ injection, LV function was determined in male Sprague-Dawley (SD) rats and TGR(hKLK1) (n=10/group) by a Millar tip catheter. Total collagen content (Sirius Red staining) and expression of types I, III, and VI collagen were quantified by digital image analysis. SD-STZ hearts demonstrated significantly higher total collagen amounts than normoglycemic controls, reflected by the concomitant increment of collagen types I, III, and VI. This correlated with a significant reduction of LV function vs. normoglycemic controls. In contrast, surface-specific content of the extracellular matrix, including collagen types I, III, and VI expression, was significantly lower in TGR(hKLK1)-STZ, not exceeding the content of SD and TGR(hKLK1) controls. This was paralleled by a preserved LV function in TGR(hKLK1)-STZ animals. The kallikrein inhibitor aprotinin and the bradykinin (BK) B2 receptor antagonist icatibant reduced the beneficial effects on LV function and collagen content in TGR(hKLK1) STZ animals. Transgenic expression of hKLK1 counteracts the progression of LV contractile dysfunction and extracellular matrix remodeling in STZ-induced diabetic cardiomyopathy via a BK B2 receptor-dependent pathway. PMID- 15117888 TI - Insulin resistance in adult cardiomyocytes undergoing dedifferentiation: role of GLUT4 expression and translocation. AB - Myocardium undergoing remodeling in vivo exhibits insulin resistance that has been attributed to a shift from the insulin-sensitive glucose transporter GLUT4 to the fetal, less insulin-sensitive, isoform GLUT1. To elucidate the role of altered GLUT4 expression in myocardial insulin resistance, glucose uptake and the expression of the glucose transporter isoforms GLUT4 and GLUT1 were measured in adult rat cardiomyocytes (ARC). ARC in culture spontaneously undergo dedifferentiation, hypertrophy-like spreading, and return to a fetal-like gene expression pattern. Insulin stimulation of 2-deoxy-D-glucose uptake was completely abolished on day 2 and 3 of culture and recovered thereafter. Although GLUT4 protein level was reduced, the time-course of unresponsiveness to insulin did not correlate with altered expression of GLUT1 and GLUT4. However, translocation of GLUT4 to the sarcolemma in response to insulin was completely abolished during transient insulin resistance. Insulin-mediated phosphorylation of Akt was not reduced, indicating that activation of phosphatidylinositol 3 kinase (PI3K) was preserved. On the other hand, total and phosphorylated Cbl was reduced during insulin resistance, suggesting that activation of Cbl/CAP is essential for insulin-mediated GLUT4 translocation, in addition to activation of PI3K. Pharmacological inhibition of contraction in insulin-sensitive ARC reduced insulin sensitivity and lowered phosphorylated Cbl. The results suggest that transient insulin resistance in ARC is related to impairment of GLUT4 translocation. A defect in the PI3K-independent insulin signaling pathway involving Cbl seems to contribute to reduced insulin responsiveness and may be related to contractile arrest. PMID- 15117889 TI - In vivo transfer of soluble TNF-alpha receptor 1 gene improves cardiac function and reduces infarct size after myocardial infarction in rats. AB - Increased circulating and cardiac TNF-alpha levels during myocardial ischemia have been found in both experimental animals and patients with ischemic heart disease and advanced heart failure. Soluble TNF-alpha receptor 1 (sTNFR1) is an antagonist to TNF-alpha. In the present study, we examined whether sTNFR1 improves cardiac function in rats after myocardial infarction. Male Wistar rats were subjected to left coronary artery (LCA) ligation. Immediately after the ligation, a total of 200 microg of either the sTNFR1 or LacZ plasmid was injected into three different sites in the left ventricular wall. From 1 to 21 days after LCA ligation, TNF-alpha bioactivity in the heart was higher in rats receiving LacZ plasmid than in sham-operated rats, whereas sTNFR1 plasmid significantly suppressed the increase. The LV diastolic dimension was significantly lower, and the fractional shortening was significantly higher in rats treated with the sTNFR1 plasmid than in those treated with the LacZ plasmid. At 21 days after LCA ligation, the LV end-diastolic pressure was also significantly lower in the rats treated with the sTNFR1 plasmid. In addition, the sTNFR1 expression plasmid had significantly reduced the infarct size. In conclusion, TNF-alpha bioactivity in the heart increased during the early stage of infarction and remained elevated. This elevation seemed partially responsible for the impairment of LV function and the increased infarct size. Suppression of TNF-alpha bioactivity from the early stage of infarction with the sTNFR1 plasmid improved cardiac function and reduced infarct size. PMID- 15117890 TI - Long-term effectiveness and safety of GnRH agonist plus raloxifene administration in women with uterine leiomyomas. AB - BACKGROUND: Our aim was to evaluate the long-term effectiveness and safety of GnRH agonist plus raloxifene administration in women with symptomatic uterine leiomyomas. METHODS: Fifty pre-menopausal women with uterine leiomyomas were treated with leuprolide acetate depot at dose of 3.75 mg/28 days and raloxifene hydrochloride at 60 mg/day for 18 cycles. At admission and after each six cycles of treatment, bone mineral density (BMD), uterine, leiomyoma and non-leiomyoma dimensions, serum bone metabolism markers, lipid, glucose and insulin levels were evaluated. Leiomyoma-related and climacteric-like symptoms were assessed using a daily diary. RESULTS: Throughout the study, no significant change in BMD or in any bone metabolism markers was observed. A significant decrease in uterine, leiomyoma and non-leiomyoma sizes was detected in comparison with baseline already after 6 months. No other significant change was observed at the successive follow-up visits. No significant change in lipid and glucose profile was detected throughout the study. The treatments were well tolerated. All treatment withdrawals (16%, eight out of 50) were due to lack of compliance, and none to drug-related adverse experiences. CONCLUSION: GnRH agonist plus raloxifene administration is an effective and safe treatment for pre-menopausal women with uterine leiomyomas. PMID- 15117891 TI - Expression of androgen receptors in upper human fetal reproductive tract. AB - BACKGROUND: Androgens play a key role in human fetal development. All androgens act through a single intracellular androgen receptor (AR), which is encoded by a single copy gene on the X chromosome. ARs are expressed as early as 9 weeks in the epithelium and mesenchyme of the urogenital sinus, paramesonephric (Mullerian) and mesonephric (Wolffian) ducts. METHODS: Using immunohistochemistry, we investigated the distribution of ARs in the gonads and lower genital tracts of 54 human fetuses at 8-11 weeks of gestation. Gender was determined by PCR. RESULTS: The AR was expressed in a similar pattern in both male and female fetuses. There appears to be no difference in expression in the mesonephros or the mesonephric ducts when male and female pelvises were compared. Expression in the female paramesonephric duct was within the epithelium, whereas, in the male pelvises, expression was in the mesenchyme of the paramesonephric duct. When AR expression was compared in the ovary and testes, both gonads seem to express AR at 9 weeks, but this expression was extended into the 10th week of gestation in the male. CONCLUSION: The specific pattern of AR expression implies a key role in gonadal development. However, the pattern of staining was similar in the gonads at 8 and 9 weeks in both sexes, although staining persisted longer in the testis until the 10th week. AR expression, therefore, is not a key determinant of human gonadal differentiation. PMID- 15117892 TI - Pregnancy following chemical activation of oocytes in a couple with repeated failure of fertilization using ICSI: case report. AB - We report our attempts to achieve a successful pregnancy outcome with calcium ionophore A23187 and puromycin oocyte activation using sperm from a normozoospermic husband of a patient with previous repeated failed fertilization following ICSI. Oocytes from the female partner of a couple with a 4 year history of unexplained primary infertility with repeated failed fertilization following ICSI were used. In the latest ICSI attempt, oocytes were activated by treatment with calcium ionophore (5 min) and puromycin (5 h), then cultured. In this cycle, assisted oocyte activation with calcium ionophore and puromycin after ICSI resulted in a satisfactory fertilization rate (8/12; 66.7%); in prior cycles only one of 71 oocytes (1.4%) was fertilized. The outcome was a Caesarean section delivery of a healthy male infant without congenital abnormalities at 41 weeks, 2 days of gestation. In conclusion, the use of calcium ionophore and puromycin for oocyte activation was found to be a useful method in a case of repeated failed fertilization after ICSI. PMID- 15117893 TI - Single top quality embryo transfer as a model for prediction of early pregnancy outcome. AB - BACKGROUND: Single embryo transfer (particularly of a top quality embryo) is an excellent model to correlate embryo quality in terms of morphological criteria to early pregnancy. We investigated whether this model could provide us with more information on what happens after implantation in the first trimester of pregnancy. METHODS: The outcome of 370 consecutive single top quality embryo transfers in patients younger than 38 years was analysed for pregnancy and first trimester pregnancy loss (FTPL) before 13 weeks of gestation. Analysis was done on each cohort of embryos from which the transferred top quality embryo was selected. Serum HCG levels were measured on day 8 and day 12 after day 3 embryo transfer. The HCG index was calculated as the level of HCG on day 12/HCG on day 8. RESULTS: The pregnancy rate after single top quality embryo transfer was 51.9%. This was independent of the patients' age. FTPL, however, appeared to be age dependent: 15.4% for the whole group, 9% in patients younger than 30 years and 19% in patients above 30 years. The pregnancy rate was 50% in IVF cycles and 52% in ICSI cycles; FTPL was 19% in IVF cycles and 10% in ICSI cycles. Multiple regression analysis showed that these differences originated from age differences between both populations rather than from technique-related factors. An HCG level >or=45 IU/l on day 12 was predictive for ongoing pregnancy with 75.6% sensitivity and 100% specificity; an HCG index >or=3.5 similarly predicted ongoing pregnancy with 72.3% sensitivity and 100% specificity. CONCLUSIONS: These data show that embryo selection for transfer on day 3 can be used as an excellent tool for prediction of pregnancy but not for prediction of FTPL. The pregnancy rate of a single top quality embryo is not related to age, whereas FTPL is age dependent. PMID- 15117894 TI - The predictive value of sperm chromatin structure assay (SCSA) parameters for the outcome of intrauterine insemination, IVF and ICSI. AB - INTRODUCTION: Sperm chromatin integrity assessment has been suggested as a fertility predictor. The aim of this study was to examine the relationship between the results of sperm chromatin structure assay (SCSA) and the outcome of IVF, ICSI and intrauterine insemination (IUI). METHODS: A total of 306 consecutive couples undergoing assisted reproduction were included. IUI was performed in 131, IVF in 109 and ICSI in 66. SCSA results were expressed as DNA fractionation index (DFI) and highly DNA stainable (HDS) cell fractions. Reproductive outcome parameters were biochemical pregnancy (BP), clinical pregnancy (CP) and delivery (D). RESULTS: For IUI, the chance of pregnancy/delivery was significantly higher in the group with DFI 27% or HDS >10%. The odds ratios (ORs) (95% confidence intervals) were 20 (2.3-117), 16 (1.9-137) and 14 (1.6-110) for BP, CP and D, respectively. No statistical difference between the outcomes of IVF versus ICSI was observed in the group with DFI 27% group, however, the results of ICSI were significantly better than those of IVF. Comparing ICSI with IVF, the OR (95% CI) for BP was 26 (1.9-350). CONCLUSIONS: SCSA is a useful method for prediction of the outcome of assisted reproduction. PMID- 15117895 TI - Detection of structural and numerical chromosomal abnormalities by ACM-FISH analysis in sperm of oligozoospermic infertility patients. AB - BACKGROUND: Modern reproductive technologies are enabling the treatment of infertile men with severe disturbances of spermatogenesis. The possibility of elevated frequencies of genetically and chromosomally defective sperm has become an issue of concern with the increased usage of ICSI, which can enable men with severely impaired sperm production to father children. Several papers have been published reporting aneuploidy in oligozoospermic patients, but relatively little is known about chromosome structural aberrations in the sperm of these patients. METHODS: We examined sperm from infertile, oligozoospermic individuals for structural and numerical chromosomal abnormalities using a multicolour ACM fluorescence in situ hybridization (FISH) assay that utilizes DNA probes specific for three regions of chromosome 1 to detect human sperm that carry numerical chromosomal abnormalities plus two categories of structural aberrations: duplications and deletions of 1pter and 1cen, and chromosomal breaks within the 1cen-1q12 region. RESULTS: There was a significant increase in the average frequencies of sperm with duplications and deletions in the infertility patients compared with the healthy concurrent controls. There was also a significantly elevated level of breaks within the 1cen-1q12 region. There was no evidence for an increase in chromosome 1 disomy, or in diploidy. CONCLUSIONS: Our data reveal that oligozoospermia is associated with chromosomal structural abnormalities, suggesting that oligozoospermic men carry a higher burden of transmissible, chromosome damage. The findings raise the possibility of elevated levels of transmissible chromosomal defects following ICSI treatment. PMID- 15117896 TI - Height, weight, and motor-social development during the first 18 months of life in 126 infants born to 109 mothers with polycystic ovary syndrome who conceived on and continued metformin through pregnancy. AB - BACKGROUND: We prospectively assessed growth and motor-social development during the first 18 months of life in 126 live births (122 pregnancies) to 109 women with polycystic ovary syndrome (PCOS) who conceived on and continued metformin (1.5-2.55 g/day) through pregnancy. METHODS: The lengths and weights of PCOS neonates were compared with gender-specific Centers for Disease Control and Prevention (CDC) infant data. Gestational diabetes (GD) and pre-eclampsia in women with PCOS were compared with 252 healthy women without PCOS who had >or=1 live birth (262 live births). RESULTS: There were 101 out of 126 (80%) term (>or=37 gestational weeks) PCOS births, which was not significantly different (P = 0.7) from controls, 206 out of 252 (81.7%). There were two (1.6%) birth defects. GD occurred in nine out of 119 PCOS pregnancies (7.6%) versus 40 out of 251 (15.9%) controls, P = 0.027. The prevalence of pre-eclampsia did not differ in PCOS versus control pregnancies (4.1 versus 3.6%, P = 0.8). The birth length and weight of the 52 male neonates did not differ (P > 0.05) from those of CDC males; the 74 female neonates were shorter than CDC females (48.9 +/- 5.4 versus 50.6 +/- 2.7 cm, P = 0.006) and weighed less (3.09 +/- 0.85 versus 3.29 +/- 0.52 kg, P = 0.04). There were no systematic differences in growth between PCOS and CDC infants over 18 months. At 3, 6, 9, 12 and 18 months, of a potential 100% motor-social development score, scores (+/-SD) were 95 +/- 13, 98 +/- 8%, 95 +/- 10, 97 +/- 8 and 94 +/- 16%; no infants had motor-social developmental delays. CONCLUSIONS: Metformin reduced development of GD, was not teratogenic and did not adversely affect birth length and weight, growth or motor-social development in the first 18 months of life. PMID- 15117897 TI - Ovarian tissue banking for cancer patients: reduction of post-transplantation ischaemic injury: intact ovary freezing and transplantation. AB - Despite reasonable achievements in different animal species, the debate about many technical aspects of ovarian tissue banking is continuing. Human ovarian tissue banks are increasingly established around the world without a clear plan about how to make the best use of such tissue. One of the important challenges facing this growing technology is to determine the ideal method for the use of this cryopreserved ovarian tissue. It is not uncommon in medicine to introduce a technology without a clear understanding of the consequences. If it is decided that ovarian tissue is to be autotransplanted, what is the most suitable place? Which technique should be implemented? As a part of the ongoing debate on ovarian tissue banking in cancer patients, this paper supports the notion that cryopreservation of an intact ovary with its vascular pedicle may be a viable alternative to the currently available techniques. Research in the development of technology to cryopreserve whole organs as well surgical techniques for the auto transplantation of an ovary with its vascular pedicle should be encouraged. PMID- 15117898 TI - The GnRH antagonist cetrorelix reduces cyclophosphamide-induced ovarian follicular destruction in mice. AB - BACKGROUND: It has remained controversial whether and in what way suppression of the pituitary-gonadal axis using GnRH analogues can reduce the destructive effects of chemotherapy on ovarian primordial follicles and thus prevent ovarian failure. GnRH antagonists suppress gonadotrophin levels immediately after administration. In this study we determined whether administration of the GnRH antagonist cetrorelix before exposure to increasing doses of cyclophosphamide (Cy) affected the number of surviving primordial follicles (PMF) in the mice ovary. METHODS: Highly inbred young Balb/c mice (114 females) were injected with 0, 50 and 75 mg/kg of Cy. In each treatment group, half of the females were injected daily with cetrorelix starting 9 days before and 7 days post administration of Cy. In serial sequential ovarian sections the total number of PMF in both ovaries was counted. RESULTS: Ovaries exposed to Cy at doses of 50 and 75 mg/kg had significantly fewer PMF than those in the control group (P < 0.01). In each of the Cy groups used, pretreatment with cetrorelix resulted in significantly higher numbers of PMF: in the 50 mg/kg Cy group only 14% were destroyed (cetrorelix group) versus 53% (P < 0.001), while in the 75 mg/kg Cy group only 35% of PMF were destroyed versus 54% in animals treated only with Cy (P < 0.004). The interaction between the effect of cetrorelix and the different doses of Cy did not reach statistical significance (P = 0.089, two-way ANOVA). CONCLUSIONS: Administration of the GnRH antagonist cetrorelix to mice significantly decreases the extent of ovarian damage induced by the chemotherapeutic agent Cy. The use of different substerilizing doses of Cy suggested that the extent of protection achieved by the antagonist is dose dependent and decreases with increasing Cy doses. The results of this study may suggest a possible similar beneficial effect in women undergoing chemotherapy, can explain the discrepancy in results of existing clinical studies and indicate possible pathways for ovarian GnRH agonist protection. Further research and clinical studies are needed in order to confirm these results. PMID- 15117899 TI - Progesterone withdrawal causes endothelin release from cultured human uterine microvascular endothelial cells. AB - BACKGROUND: Current theories on the physiology of menstrual bleeding in humans offers an explanation for the shedding of the endometrium as a result of a breakdown of the extracellular matrix due to an inflammatory reaction. The link between the fall in progesterone levels and these events is not clear. Neither has an explanation been presented for the vasoconstriction in the coiled arteries occurring during menses. We have hypothesized a chain of events where the fall in progesterone levels induces an upregulation of the thrombin receptor in the small uterine arteries leading to an increased thrombin response and subsequent endothelin release. METHODS: Endothelial cells from human umbilical cord (HUVECs) and from human small uterine arteries (UtMVECs) were cultured under conditions partly resembling the female hormonal cycle with progesterone withdrawal. RESULTS: Following progesterone increase and subsequent withdrawal, we found an increased production of thrombin receptor and an increased release of endothelin from UtMVECs compared with HUVECs. CONCLUSION: Endothelin release in response to progesterone withdrawal in UtMVECs can offer an explanation for the vasoconstriction seen in the coiled arteries during menses in humans. PMID- 15117900 TI - Suppression of the high endogenous levels of plasma FSH in infertile men are associated with improved Sertoli cell function as reflected by elevated levels of plasma inhibin B. AB - BACKGROUND: In vitro continuous stimulation of Sertoli cells with FSH leads to a desensitization of these cells to FSH action. To evaluate the presence of a desensitization of FSH receptor on Sertoli cells in vivo, we performed a controlled clinical study in 97 men affected by severe oligozoospermia. METHODS: On the basis of FSH and inhibin B plasma concentrations, these subjects were divided into three groups: group A, 33 subjects with high FSH and low inhibin B plasma levels; group B, 32 subjects with high FSH plasma levels and inhibin B concentrations at the lower limit of the normal range; and group C, 32 subjects with normal FSH and inhibin B plasma levels. Patients with high FSH plasma levels (groups A and B) were prospectively randomized into two subgroups, called A1, A2, B1 and B2. Patients of groups A1 and B1 were treated with a GnRH agonist, leuprolide acetate, to induce a hypogonadotrophic state and then were treated with recombinant human FSH (r-hFSH; 100 IU/day) and hCG (2000 IU/twice a week) for 2 months. Subjects of groups A2, B2 and C were treated only with r-hFSH for the same period. RESULTS: In patients of group A1, inhibin B remained unmodified during the whole period of study, whereas in subjects of group B1, we observed a significant reduction of this hormone during the hypogonadotrophic period and then an increase of inhibin B plasma levels that were higher that those observed before therapy. In patients of groups A2 and B2, FSH treatment did not induce a significant increase in inhibin B concentrations. In patients of group C, FSH induced a significant increase in inhibin B plasma levels. CONCLUSIONS: In infertile men, suppression of the high endogenous levels of plasma FSH associated with much lower exogenous FSH levels is able to evoke higher inhibin B production, which may indicate improved Sertoli cell function and the possibility that this could have a positive effect on spermatogenesis. PMID- 15117901 TI - Associations between androgen receptor CAG repeat length and sperm morphology. AB - BACKGROUND: The number of (trinucleotide) CAG repeats within the androgen receptor (AR) gene is inversely correlated with transcriptional activity of testosterone-target genes. Although abnormally long CAG repeats are strongly associated with male infertility, it is unclear whether CAG repeat length polymorphism can affect androgen receptor activity and sperm parameters. To explore the previously suggested association between CAG repeats and male fertility, we conducted this prospective cohort study. METHODS: We enrolled 172 men attending the IVF unit in Shaare-Zedek Medical Center. Sperm concentration, motility and morphology and the number of CAG repeats in the AR gene were measured. RESULTS: Mean CAG repeat length was greater in teratozoospermia (<14% normal forms, strict criteria) than in the normal morphology group [mean +/- 95% confidence interval (CI) 22.19 +/- 0.38 versus 21.25 +/- 0.28, P = 0.02]. Logistic regression models showed that longer CAG repeats were associated with abnormal sperm morphology [odds ratio for percentage of normal forms per unit increase in CAG repeat length 1.14 (95% CI 1.01-1.28), P = 0.04]. No association was found between CAG repeat length and sperm concentration or motility. CONCLUSIONS: We found a positive correlation between CAG repeat length and teratozoospermia. This finding validates the concept that AR function is inversely regulated by length of its CAG repeat tract. PMID- 15117902 TI - Metformin treatment before IVF/ICSI in women with polycystic ovary syndrome; a prospective, randomized, double blind study. AB - BACKGROUND: Our aim was to investigate the effect of pre-treatment with metformin in women with polycystic ovary syndrome (PCOS) scheduled for IVF stimulation. METHODS: Seventy-three oligo/amenorrhoeic women with polycystic ovaries and at least one of the following criteria: hyperandrogenaemia, elevated LH/FSH ratio, hyperinsulinism, decreased SHBG levels or hirsutism, were studied. Normal weight and overweight patients were randomized separately in a prospective, randomized, double blind study. All patients were treated for at least 16 weeks with metformin (1000 mg bid) or placebo ending on the day of HCG injection. RESULTS: No differences were found in the primary end-points: duration of FSH stimulation 14.4 (13.1-15.7) versus 14.2 (12.6-15.7) days or estradiol on the day of HCG injection 6.8 (5.3-8.2) versus 7.6 (5.6-9.6) nmol/l in the metformin and placebo groups, respectively. The secondary end-points number of oocytes, fertilization rates, embryo quality, pregnancy rates and clinical pregnancy rates were equal. However, in the normal weight subgroup (BMI <28 kg/m(2), n = 27), pregnancy rates following IVF were 0.71 (0.63-0.79) versus 0.23 (0.15-0.31) in the metformin and placebo groups, respectively (P = 0.04). Overall clinical pregnancy rates were equal: 0.51 (0.34-0.68) versus 0.44 (0.27-0.62) in the metformin and placebo groups, respectively. However, in the normal weight subgroup, clinical pregnancy rates were 0.67 (0.43-0.91) and 0.33 (0.06-0.60), respectively (P = 0.06). CONCLUSIONS: Pre-treatment with metformin prior to conventional IVF/ICSI in women with PCOS does not improve stimulation or clinical outcome. However, among normal weight PCOS women, pre-treatment with metformin tends to improve pregnancy rates. Further studies in subgroups of PCOS women are required. PMID- 15117903 TI - Recovery of fertility after grafting of cryopreserved germinative tissue in female rabbits following radiotherapy. AB - BACKGROUND: Many cancer survivors face infertility as a consequence of the aggressive treatment they must undergo. Cryopreservation of ovarian tissue before chemotherapy or radiotherapy may allow for tissue transplantation after the treatment, and restoration of fertility. We tested the potential of an orthotopic autografting of cryopreserved germinative tissue in female rabbits with ovarian failure following radiotherapy. METHODS: Ten adult multiparous female rabbits were randomly allocated into two groups, five in group I (control) and five in group II (transplant). All rabbits underwent right oophorectomy with cryopreservation of the germinative tissue, followed by sterilization of the remaining left ovary by radiotherapy. Later, group II rabbits received in the irradiated left ovary an implant of the frozen germinative tissue from the right ovary, whose small pieces were freely spread intracortically in a procedure we named 'intracortical sowing of germinative tissue' (ISGT). RESULTS: All group II rabbits conceived following spontaneous mating within 6 months of the transplant, whereas none of the remaining rabbits in group I had conceived up to 11 months after transplant. CONCLUSIONS: This study suggests that fertility can be restored in rabbits by sowing cortical tissue in a previously irradiated ovary. The clinical feasibility of this technique remains to be determined. PMID- 15117904 TI - Full-term pregnancies achieved with ICSI despite high levels of sperm chromatin damage. AB - BACKGROUND: Sperm DNA integrity is essential for the accurate transmission of genetic information. The clinical significance of this assessment lies in its association with not only natural conception rates, but also the success of assisted reproduction technology (ART). It has been reported that sperm chromatin structure assay (SCSA) identified thresholds for negative pregnancy outcome after ART when the DNA fragmentation index (DFI), previously known as COMPalphat, was >30%. METHODS: In a prospective clinical study, we examined 34 male infertile patients, the husbands of women undergoing conventional IVF or ICSI. SCSA and ART were carried out on semen aliquots taken from the same ejaculate. Fertilization rate, embryo quality and pregnancy rates were correlated to SCSA parameters, DFI and highly DNA stainable (HDS) cells. RESULTS: No differences were seen in SCSA parameter values between patients initiating pregnancies and not doing so in either ICSI or conventional IVF. Pregnancies and normal delivery were obtained even with high levels of DFI. CONCLUSIONS: There is still controversy over whether analytical techniques currently in use are able to identify the level of damage to spermatozoa. Large-scale studies should be conducted in different clinical settings to determine the effects of sperm DNA damage on the outcome of ART. PMID- 15117905 TI - Sperm FISH studies in seven male carriers of Robertsonian translocation t(13;14)(q10;q10). AB - BACKGROUND: Robertsonian translocation t(13;14) is one of the most common structural reorganization in humans, but meiotic segregation studies in these carriers are still limited. The segregation pattern of the chromosomes involved, the possible influence of the translocated chromosomes on the synapsis and disjunction of other chromosome pairs [interchromosomal effects (ICE)] and the rates of unbalanced spermatozoa produced still deserve attention, not only to obtain a better characterization of the meiotic behaviour of this reorganization, but also to offer carrier couples accurate genetic counselling. METHODS: Multicolour fluorescence in-situ hybridization was used to analyse the segregation of chromosomes 13 and 14 and the possible occurrence of ICE (on chromosomes 18, 21, 22, X and Y) in seven male carriers of a t(13;14)(q10;q10). RESULTS AND CONCLUSIONS: The individuals analysed showed a homogeneous segregation pattern, with a clear predominance of alternate segregations resulting in the production of normal/balanced spermatozoa (83-88.23%). A significant increase in the disomy rates for the sex chromosomes, which could be considered as a positive ICE, was observed in two of the carriers analysed. PMID- 15117906 TI - An algorithm to predict pregnancy in assisted reproduction. AB - BACKGROUND: Male fertility potential cannot be measured by conventional parameters for the assisted reproduction technique; ICSI. This study determines the relationship between testicular and ejaculated sperm mitochondrial (mt) DNA deletions, nuclear (n) DNA fragmentation, and fertilization and pregnancy rates in ICSI. METHODS: Ejaculated sperm were obtained from 77 men and testicular sperm from 28 men with obstructive azoospermia undergoing ICSI. Testicular sperm were retrieved using a Trucut needle. mtDNA was analysed using a long PCR. The alkaline Comet assay determined nDNA fragmentation. RESULTS: Of subjects who achieved a pregnancy (50%) using testicular sperm, only 26% had partners' sperm with wild-type (WT) mtDNA. Of pregnant subjects (38%) using ejaculated sperm, only 8% had partner sperm with WT mtDNA. In each, the successful group had less mtDNA deletions and less nDNA fragmentation. There were inverse relationships between pregnancy and mtDNA deletion numbers, size and nDNA fragmentation for both testicular and ejaculated sperm. No relationships were observed with fertilization rates. An algorithm for the prediction of pregnancy is presented based on the quality of sperm nDNA and mtDNA. CONCLUSION: In both testicular and ejaculated sperm, mtDNA deletions and nDNA fragmentation are closely associated with pregnancy in ICSI. PMID- 15117907 TI - Pesticide exposure in male farmers and survival of in vitro fertilized pregnancies. AB - BACKGROUND: Male-mediated spontaneous abortion is well described among animals, but less well documented in humans. Studies that include early pregnancy experience are expensive to conduct, but IVF provides new opportunities. MATERIALS AND METHODS: The Danish IVF register covers all IVF treatments performed after 1993. We mailed a questionnaire to 5879 women (response rate 68.2%). A subgroup of exposed male partners received a questionnaire on specific exposure to pesticides and growth retardants (n = 128 men, response rate 81.3%). Information on outcome was collected from national health registers. Survival of the first HCG-positive pregnancy was analysed using Cox regression. RESULTS: The proportion of pregnancies terminated by spontaneous abortion before 28 gestational weeks was 19.7% (n = 66 pregnancies), 19.7% (n = 61), 21.3% (n = 47) and 22.2% (n = 18) in pregnancies with paternal exposure to herbicides, fungicides, pesticides and growth retardants. respectively. In the reference group of 2925 pregnancies, the abortion rate was 28.4%. The differences in survival were not statistically significant. CONCLUSIONS: We found no increased risk of spontaneous abortion in IVF-treated women attributable to paternal agricultural application of pesticides and growth retardants. Exposure to potentially harmful pesticides in Denmark is relatively low, and the findings are restricted to countries with similar standards of protection. PMID- 15117908 TI - Blood pressure control and physician management of hypertension in hospital hypertension units in Spain. AB - Goal blood pressure (BP) was defined by the Sixth Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC-VI) and the World Health Organization-International Society of Hypertension (WHO/ISH) as <140 mm Hg systolic and <90 mm Hg diastolic for the general and <130 mm Hg systolic and <85 mm Hg diastolic for special high-risk populations. However, there are few reports that address BP control among special subgroups of hypertensives by reference to targeted BP. We therefore conducted a study to evaluate BP control of 4049 hypertensives in 47 hospital-based hypertension units in Spain. Overall, 42% of patients achieved goal BP (<140 mm Hg systolic and <90 mm Hg diastolic). Only 13% of diabetic patients and 17% of those with renal disease achieved the BP goal (<130 mm Hg systolic and <85 mm Hg diastolic), and only 10% and 12%, respectively, achieved the even more rigorous goal (<130 mm Hg systolic and <80 mm Hg diastolic). Likewise, only 18% of patients in JNC-VI risk group C and 17% of WHO/ISH high-risk patients attained a goal BP <130 mm Hg systolic and <85 mm Hg diastolic. BP control (<125 mm Hg systolic and <75 mm Hg diastolic) was extremely low (2%) in patients with proteinuria >1 g/d. Poorer BP control was observed among patients at high risk, with diabetes, renal disease, or obesity, than in lower-risk groups. BP control was lower for systolic than for diastolic BP. In >50% of uncontrolled patients, no measures were taken by doctors to optimize pharmacologic treatment, and approximately one-third of patients were still using drug monotherapy. Control of BP, particularly of systolic BP, is still far from optimal in hospital-based hypertension units. Patients at high risk, with diabetes or proteinuria, warrant focused attention. Moreover, a more aggressive behavior of doctors treating uncontrolled hypertension is needed. PMID- 15117909 TI - Reduced uteroplacental blood flow alters renal arterial reactivity and glomerular properties in the rat offspring. AB - Fetal malnutrition and hypoxia may modify organ system maturation and result in cardiovascular diseases in the adult. We tested whether intrauterine stress (IUS) leads to persistent alterations of renal biology. In rats, intrauterine stress was induced by ligation of the uterine arteries at day 17 of pregnancy. Renal arteries of the 21-day-old male offspring were isolated to study pharmacological reactivity. Kidneys were dissected to analyze renal structure and beta adrenoceptor expression. At 21 days of age, half of the animals underwent unilateral left nephrectomy. At the age of 12 weeks, rats were instrumented for blood pressure monitoring, blood sampling, and renal function measurements. After IUS, litter size and birth weight were reduced, whereas the hematocrit was increased. Renal arterial responses to beta-adrenergic stimulation and sensitivity to adenylyl cyclase activation were increased, along with the renal expression of beta2-adrenoceptors. At 21 days and at 6 months of age, the number and density of the glomeruli were reduced, whereas their size was increased. The filtration fraction and urinary albumin concentration were increased 12 weeks after intrauterine stress. In control rats, removal of the left kidney at 21 days of age did not affect kidney function and blood pressure. However, after IUS, the remaining right kidney failed to compensate for the loss of the left kidney, and blood pressure was increased. In conclusion, prenatal stress transiently modifies renal arterial reactivity and results in long-lasting adverse effects on renal structure and function and on renal compensatory mechanisms. PMID- 15117910 TI - Homocysteine enhances endothelial apoptosis via upregulation of Fas-mediated pathways. AB - Hyperhomocysteinemia is an independent risk factor for the development of atherosclerosis. However, the underlying mechanism of endothelial cell injury in hyperhomocysteinemia has not been elucidated. In this study, we examined the effect of homocysteine (Hcy) on Fas-mediated apoptosis in endothelial cells. Hcy induced upregulation of Fas in endothelial cells (ECs) in a dose-dependent manner. At the same time, Hcy increased intracellular peroxide in ECs. Hcy induced Fas expression was inhibited by the treatment with catalase. Hcy increased NF-kappaB DNA binding activity, and adenovirus-mediated transfection of a Ikappa-B mutant (Ikappa-B mt) gene inhibited Hcy-induced Fas expression. ECs were sensitive to Fas-mediated apoptosis when exposed to Hcy. Under these condition, Ikappa-B mt protected ECs from Fas-mediated apoptosis. In addition, Hcy inhibited expression of the caspase-8 inhibitor FLICE-inhibitory protein (FLIP). Adenovirus-mediated transfection of constitutively active Akt gene abolished the Hcy-mediated downregulation of FLIP. These data suggest that upregulation of Fas expression and downregulation of FLIP is a mechanism through which Hcy induces EC apoptosis. PMID- 15117911 TI - Variability in response to antihypertensive drug treatment. PMID- 15117912 TI - Determinants of blood pressure response to quinapril in black and white hypertensive patients: the Quinapril Titration Interval Management Evaluation trial. AB - Race has been considered an important factor in determining blood pressure response to treatment and selection of antihypertensive drug therapy. Data collected during a clinical trial that evaluated rapidity of medication up titration with blood pressure response to monotherapy with the angiotensin converting enzyme (ACE) inhibitor quinapril were used to characterize response in 533 black and 2046 white participants. Our objectives were to examine the influence of race and other factors on blood pressure response and to assess the degree to which nonrace factors account for apparent racial differences in response. Average systolic and diastolic blood pressure responses (baseline minus follow-up) to treatment were assessed with treatment groups combined. Crude systolic and diastolic blood pressure responses averaged 4.7 and 2.4 mm Hg less, respectively, in black compared with white participants; however, the response distributions largely overlapped. In multivariate linear regression models adjusted for study design variables and measured participant characteristics, the racial difference in systolic response was reduced by 51% to 2.3 mm Hg, and diastolic response by 21% to 1.9 mm Hg. In these models, participant characteristics, including age, gender, body size, and pretreatment blood pressure severity, significantly predicted either attenuated or enhanced blood pressure response to treatment. Our findings demonstrate that a large source of variability of blood pressure response to treatment is within, not between, racial groups, and that factors that vary at the level of the individual contribute to apparent racial differences in response to treatment. PMID- 15117914 TI - Geneticism of essential hypertension. PMID- 15117913 TI - Eplerenone prevents salt-induced vascular remodeling and cardiac fibrosis in stroke-prone spontaneously hypertensive rats. AB - We examined the effect of different levels of salt intake on the role of aldosterone on cardiac and vascular changes in salt-loaded stroke-prone spontaneously hypertensive rats (SHRSP). Eleven-week-old SHRSP were fed high-salt (4.2% NaCl), normal-salt (0.28%), or low-salt (0.03%) diets with or without eplerenone (100 mg/kg per day, in food) for 5 weeks. A group of high-salt SHRSP was also treated with hydralazine (25 mg/kg per day). Blood pressure increased more in high-salt rats than in other groups (P<0.001). Eplerenone prevented further blood pressure rise in salt-loaded rats, with little effect on control and low-salt SHRSP. Increased media-to-lumen ratio of mesenteric resistance arteries induced by salt (P<0.01) was prevented by eplerenone (P<0.01). Maximal acetylcholine-induced vasodilation was impaired under salt loading (P<0.01), but improved under eplerenone (P<0.01). Eplerenone prevented (P<0.01) increased heart weight and left and right ventricular collagen deposition induced by high salt. Blood pressure lowering by hydralazine in high-salt SHRSP did not influence endothelial function or left ventricular collagen. Our study demonstrates salt dependency of aldosterone effects on severity of hypertension, endothelial dysfunction, and cardiac and vascular remodeling in SHRSP. These effects were attenuated by eplerenone, particularly in the salt-loaded state, underlining the pathophysiological role of aldosterone in salt-sensitive hypertension. PMID- 15117916 TI - Tooth loss is associated with an increased risk of hypertension in postmenopausal women. AB - Tooth loss has been associated with an increased risk of vascular diseases such as coronary heart disease and cerebrovascular disease. Little is known whether hypertension is an important factor linking 2 phenomena in postmenopausal women. We compared an incidence of hypertension and traditional risk factors for vascular diseases between 2 age-matched groups: 67 postmenopausal women with missing teeth and 31 without missing teeth. In addition to blood pressure, serum concentration of total cholesterol, high- and low-density lipoprotein cholesterol and triglycerides, plasma angiotensin-converting enzyme activity, plasma angiotensin II concentration, plasma renin activity, and resting heart rate were measured as traditional risk factors for vascular diseases. Subjects without missing teeth had significantly lower diastolic blood pressure than did subjects with missing teeth (P=0.021). The former tended to have lower systolic blood pressure than did the latter (P=0.058). There were no significant differences in other variables between subjects with and without missing teeth. The odds ratio of having hypertension in subjects with missing teeth was 3.59 (95% confidence interval, 1.10 to 11.7) after adjustment of obesity, hypercholesterolemia, and hypertriglyceridemia. Our results suggest that hypertension may be an important factor linking tooth loss and an increased risk of vascular diseases in postmenopausal women. PMID- 15117915 TI - Blood pressure differences between northern and southern Chinese: role of dietary factors: the International Study on Macronutrients and Blood Pressure. AB - Blood pressure and prevalence of high blood pressure are greater for northern than southern Chinese. Reasons for these differences are unclear. Relationships of north-south blood pressure differences with multiple dietary factors were investigated in 839 Chinese participants, International Study on Macronutrients and Blood Pressure (INTERMAP), 561 northern, 278 southern, aged 40 to 59 years. Daily nutrient intakes were determined from four 24-hour dietary recalls and 2 timed 24-hour urine collections. Average systolic/diastolic pressure levels were 7.4/6.9 mm Hg higher for northern than southern participants. Southern participants had lower body mass index, sodium intake, sodium/potassium ratio, and higher intake of calcium, magnesium, phosphorus, and vitamins A and C. Considered singly, with control for age and gender, several dietary variables (eg, body mass index, urinary sodium/potassium ratio, urinary sodium, dietary phosphorus, and magnesium) reduced north-south blood pressure differences by > or =10%. Controlled for age and gender, nondietary variables had little effect on north-south blood pressure differences. With inclusion in regression models of multiple dietary variables (sodium, potassium, magnesium or phosphorus, body mass index), north-south blood pressure differences became much smaller (systolic 1.1, diastolic 1.6 mm Hg) and statistically nonsignificant. In conclusion, multiple dietary factors accounted importantly for north-south blood pressure differences. Efforts are needed to improve nutrition in China, especially in the north, as well as in other populations including those in the United States, for prevention and control of adverse blood pressure levels and major adult cardiovascular disease. PMID- 15117917 TI - Cardiac angiotensin AT2 receptor: what exactly does it do? PMID- 15117918 TI - Profibrotic effects of angiotensin II in the heart: a matter of mediators. PMID- 15117919 TI - Susceptibility of porcine cytomegalovirus to antiviral drugs. AB - OBJECTIVES: Re-activation of porcine cytomegalovirus (PCMV) in the xenograft has been reported in pig-to-baboon models of xenotransplantation and is associated with invasive disease and consumptive coagulopathy. If xenotransplantation of porcine organs into human recipients is to proceed, donor organs will have to be free from a wide range of infectious agents including PCMV. However, it is prudent to characterize the antiviral susceptibility of this virus. We therefore investigated the effect of selected antiviral agents, currently licensed for the treatment of human herpesvirus infections, on PCMV replication. METHODS: Antiviral susceptibility was determined using real-time PCR and indirect immunofluorescence measurements in a porcine fallopian tube cell line infected with PCMV. RESULTS: PCMV replication was significantly inhibited by ganciclovir and cidofovir (both EC(50) < 1 mg/L) and to a lesser extent by foscarnet (EC(50) within range 25-50 mg/L) and aciclovir (EC(50) > 25 mg/L). CONCLUSIONS: These results show that, if it proves necessary, ganciclovir and cidofovir should be considered as first-line drugs to treat PCMV infections in xenograft recipients. PMID- 15117920 TI - Defined daily doses of antimicrobials reflect antimicrobial prescriptions in ambulatory care. AB - BACKGROUND: Data on actual antimicrobial prescriptions are rarely made available and the World Health Organization (WHO) recommends using the number of defined daily doses (DDDs) as a measurement unit for inter-country comparisons of antimicrobial use in outpatients. However, because antimicrobials are given for limited periods of time and the DDDs may not represent the doses used in practice, this measurement unit has been blamed for poorly reflecting the number of antimicrobial prescriptions in outpatients. METHODS: For 11 out of 15 European countries, data on national outpatient sales of antimicrobials and on antimicrobial prescriptions to outpatients in 1997 were purchased from the Institute for Medical Statistics (IMS) Health. For two additional countries, i.e. Sweden and Denmark, similar data were obtained from the National Corporation of Swedish Pharmacies and the Danish Medicines Agency, respectively. The relationships between the number of DDDs and the number of prescriptions, on the one hand, and the antimicrobial use density (DDD per 1000 inhabitant-days) and the prescription rate (prescriptions per 1000 inhabitants), on the other hand, were assessed with the two-tailed Spearman coefficient for non-parametric correlations. RESULTS: The number of DDDs, as defined by the WHO, and the number of prescriptions of antimicrobials to outpatients in European countries in 1997 were strongly correlated. Similarly, the antimicrobial use density and the prescription rate in these countries were strongly correlated. These relationships were found for total systemic antimicrobials and for all major antimicrobial classes. CONCLUSION: Our results confirm the relevance of the number of DDDs per 1000 inhabitant-days as a measurement unit to compare outpatient antimicrobial use among countries or regions. PMID- 15117921 TI - Characterization of the molecular mechanisms of quinolone resistance in Yersinia enterocolitica O:3 clinical isolates. AB - OBJECTIVES: The aim of this study was to determine the roles of mutations in the gyrA and parC genes and the overexpression of efflux pump(s) as mechanisms of resistance to quinolones. Forty-five Yersinia enterocolitica O:3 clinical isolates (41 nalidixic acid-resistant, three nalidixic acid-susceptible and one nalidixic acid-resistant strain obtained in vitro) were analysed. RESULTS: All the nalidixic acid-resistant strains showed mutations in the gyrA gene and none in the parC gene. The presence of the inhibitor produced decreases in the MIC values of nalidixic acid by two to six serial dilution steps in 37 of the 41 nalidixic acid-resistant strains. Meanwhile, the MIC value of ciprofloxacin was affected in two strains whose values diminished three serial dilution steps. The nalidixic acid-resistant mutant obtained in vitro was also affected by the inhibitor decreasing the MIC value of nalidixic acid three serial dilutions steps whereas the MICs for the nalidixic acid-susceptible strains were not affected. CONCLUSIONS: Our results show that the high level of resistance to nalidixic acid is likely due to an overexpression of an efflux pump plus a mutation in the gyrA gene, whereas decreased susceptibility to ciprofloxacin is only associated with the presence of a mutation in the gyrA gene. PMID- 15117922 TI - Synthetic furanones inhibit quorum-sensing and enhance bacterial clearance in Pseudomonas aeruginosa lung infection in mice. AB - INTRODUCTION: Antibiotics are used to treat bacterial infections by killing the bacteria or inhibiting their growth, but resistance to antibiotics can develop readily. The discovery that bacterial quorum-sensing regulates bacterial virulence as well as the formation of biofilms opens up new ways to control certain bacterial infections. Furanone compounds capable of inhibiting bacterial quorum-sensing systems have been isolated from the marine macro alga Delisea pulchra. OBJECTIVES: Two synthetic furanones were tested for their ability to attenuate bacterial virulence in the mouse models of chronic lung infection by targeting bacterial quorum-sensing without directly killing bacteria or inhibiting their growth. METHODS: Study I. Mice with Escherichia coli MT102 [luxR PluxI-gfp(ASV)] lung infection were injected intravenously with N-acyl homoserine lactones with or without furanones to test the interference of furanones with quorum-sensing. Study II. Mice with lung infection by Pseudomonas aeruginosa PAO1 [dsred, lasR-PlasB-gfp(ASV)] were injected intravenously with furanones to evaluate their inhibiting effects on quorum-sensing. Study III. Mice with P. aeruginosa PAO1 lung infection were treated with different doses of furanones to evaluate the therapeutic effects of furanones on the lung infection. RESULTS: Furanones successfully interfered with N-acyl homoserine lactone and suppressed bacterial quorum-sensing in lungs, which resulted in decreases in expression of green fluorescent protein. Furanones accelerated lung bacterial clearance, and reduced the severity of lung pathology. In a lethal P. aeruginosa lung infection, treatment with furanone significantly prolonged the survival time of the mice. CONCLUSION: Synthetic furanone compounds inhibited bacterial quorum-sensing in P. aeruginosa and exhibited favourable therapeutic effects on P. aeruginosa lung infection. PMID- 15117923 TI - New aminoglycoside acetyltransferase gene, aac(3)-Id, in a class 1 integron from a multiresistant strain of Vibrio fluvialis isolated from an infant aged 6 months. AB - OBJECTIVES: To characterize the molecular basis of antibiotic resistance in a multidrug-resistant clinical isolate of Vibrio fluvialis H-08942. PATIENT AND METHODS: V. fluvialis H-08942 was isolated from a hospitalized infant aged 6 months suffering from cholera-like diarrhoea in India in 2002. The broth microdilution method was used to determine the MICs of a range of antibiotics for this strain. PCR, DNA sequencing, Southern hybridization, cloning and expression were used to characterize the molecular basis of antibiotic resistances. RESULTS: V. fluvialis H-08942 showed resistance to chloramphenicol, streptomycin, spectinomycin, co-trimoxazole, ampicillin, furazolidone, nalidixic acid and gentamicin. A class 1 integron that contains a novel aminoglycoside acetyltransferase gene, aac(3)-Id, and aminoglycoside adenyltransferase gene, aadA7, was characterized. The aac(3)-Id gene product was found to share 50%, 45% and 44% identity to AAC(3)-Ic, AAC(3)-Ia, and AAC(3)-Ib, respectively. Both aac(3)-Id and aadA7 genes were cloned and expressed in Escherichia coli. Phylogenetic analysis suggested that the aac(3)-Id represents a fourth evolutionary lineage in the aminoglycoside acetyltransferase genes. Southern hybridization showed that this integron is located in the chromosome. CONCLUSIONS: In this study we identified a new type of aminoglycoside acetyltransferase gene, aac(3)-Id. In addition, this is the first report of identification of antibiotic resistance genes and a class 1 integron in V. fluvialis. PMID- 15117924 TI - Antimicrobial susceptibility patterns and macrolide resistance genes of viridans group streptococci from blood cultures in Korea. AB - OBJECTIVES: Our aim was to study the antimicrobial susceptibilities and macrolide resistance mechanisms of viridans group streptococci (VGS) in a Korean tertiary hospital. METHODS: MICs of five antimicrobials were determined for 106 VGS isolated from blood cultures. The macrolide resistance mechanisms of erythromycin non-susceptible isolates were studied by the double-disc test and PCR. RESULTS: In all, 42.4% of the isolates were susceptible to penicillin. Nine of 61 penicillin non-susceptible isolates were fully resistant (MIC >/= 4 mg/L). Rates of non-susceptibility to erythromycin, clindamycin and ceftriaxone were 33.9%, 17.9% and 9.4%, respectively. Twenty-two (61.1%) of 36 erythromycin non susceptible isolates expressed constitutive resistance to macrolide-lincosamide streptogramin B antibiotics (a constitutive MLS(B) phenotype); 13 isolates (36.1%) expressed an M phenotype; and one isolate, a Streptococcus bovis isolate, had an inducible MLS(B) resistance phenotype. erm(B) was found in isolates with constitutive/inducible MLS(B) phenotypes, and mef(A) in isolates with the M phenotype. In three isolates (two isolates with a constitutive MLS(B) phenotype and in one isolate with the M phenotype), none of erm(A), erm(B), erm(C) or mef(A) was detected by PCR. CONCLUSIONS: Penicillin non-susceptible VGS were more resistant to erythromycin, clindamycin and ceftriaxone than were penicillin susceptible isolates. A constitutive MLS(B) phenotype associated with erm(B) was the predominant mechanism of macrolide resistance among erythromycin non susceptible isolates from this Korean hospital. PMID- 15117925 TI - In vitro susceptibility of recent antibiotic-resistant urinary pathogens to ertapenem and 12 other antibiotics. AB - BACKGROUND: The treatment of complicated urinary tract infections may require the use of a parenteral antibiotic with potent activity against the most common urinary pathogens. Ertapenem is a broad-spectrum 1beta-methyl carbapenem with a long plasma half-life that allows administration of a single daily dose. METHODS: The purpose of this work was to test the in vitro susceptibility to ertapenem, ampicillin, cefazolin, cefuroxime, cefotaxime, co-amoxiclav, piperacillin/tazobactam, imipenem, gentamicin, amikacin, fosfomycin, ciprofloxacin and co-trimoxazole of 482 strains of urinary pathogens of the family Enterobacteriaceae isolated from patients in the community of Madrid (40% from males). The distribution was as follows: Escherichia coli (n = 315), Proteus mirabilis (n = 42), Klebsiella spp. (n = 14) and AmpC-producing Enterobacteriaceae (n = 111). The strains studied were selected based on their resistance to quinolones and aminoglycosides, and their production of extended spectrum beta-lactamases (ESBLs) or AmpC-type beta-lactamases. RESULTS: All the strains were susceptible to ertapenem, imipenem and amikacin. The MIC(90) of ertapenem ranged from a minimum of 0.03 mg/L for Proteus vulgaris and a maximum of 1 mg/L for Enterobacter spp. Ertapenem was the most active of all drugs tested in all cases. On comparing antibiotic resistance among ESBL-producing strains of E. coli (n = 35) and E. coli strains not producing ESBLs (n = 280), statistically significant differences were obtained for ciprofloxacin (P = 0.002) and gentamicin (P = 0.011). Regarding ertapenem, only a slight increase in MIC(50) was seen, the value being 0.015 mg/L for strains not producing ESBLs versus 0.03 mg/L for ESBL-producing strains. CONCLUSIONS: In view of its significant antibiotic potency against antibiotic-resistant Enterobacteriaceae, ertapenem may constitute a good therapeutic alternative in urinary infections caused by these pathogens. PMID- 15117926 TI - Antimicrobial activity of clofazimine is not dependent on mycobacterial C-type phospholipases. AB - We have used a phospholipase C (PLC)-deletion mutant (plcABC) of the H37Rv strain of Mycobacterium tuberculosis (MTB), as well as a plcA-insertion mutant of Mycobacterium smegmatis, to investigate the possible involvement of PLCs in clofazimine-mediated inhibition of mycobacterial K(+) transport and growth. Inactivation of the PLCs of MTB and insertion of the plcA gene into M. smegmatis resulted in a substantial reduction and increase in hydrolysis of phosphatidylcholine (PC), respectively. However, both the mutant and wild-type strains of MTB and M. smegmatis were equally sensitive to the inhibitory effects of clofazimine on K(+) uptake and growth. These observations demonstrate that the PLCs of MTB are not involved in the antimicrobial activity of clofazimine. PMID- 15117928 TI - Interaction between HIV-1 and HCV infections: towards a new entity? AB - Since human immunodeficiency virus (HIV) and hepatitis C virus (HCV) share the same modes of transmission, HIV-HCV co-infected patients are relatively common. Until recently, the clinical course of HCV in co-infected patients was overshadowed by the high morbidity and mortality of HIV disease. Recent reductions in morbidity and mortality among HIV-infected patients due to the advent of highly active antiretroviral therapy (HAART), have contributed to the emergence of HCV as a significant viral pathogen in this population. This article reviews the current evidence on the epidemiology and clinical implications of an interaction between HIV-1 and HCV infections. PMID- 15117927 TI - Treatment of acute Chlamydia pneumoniae infection with telithromycin in C57BL/6J mice. AB - OBJECTIVES: The efficacy of telithromycin, a new ketolide antibiotic, was investigated in the treatment of acute Chlamydia pneumoniae infection in a mouse model. METHODS: C57BL/6J mice were inoculated intranasally, and the effects of three different doses of telithromycin (25, 50 and 100 mg/kg) were assessed after 5 and 10 days of treatment. Lungs for culture, PCR, histopathology, and blood for serum samples were collected immediately after each treatment period and at 3 weeks post-inoculation. C. pneumoniae-specific antibodies were analysed, and the effect of treatment was assessed by culture, detection of C. pneumoniae DNA and determination of histopathological inflammatory changes in mouse lungs. RESULTS: Culture negativity in the lungs was achieved with the higher doses, 50 and 100 mg/kg, after 10 days of treatment. C. pneumoniae DNA was not totally eradicated with the treatments, but the groups treated with 50 and 100 mg/kg doses for 10 days had the lowest DNA positivity rates (10%) 3 weeks after the inoculation. In lung histopathology, the efficacy of telithromycin on inflammatory changes was also dose-dependent: higher doses were more effective in reducing the inflammatory reaction. Overall, the 25 mg/kg dose had a weaker effect compared with the others. CONCLUSIONS: Telithromycin had both time- and dose-dependent effects on the eradication of chlamydia and on reducing infection-induced inflammatory changes in mouse lungs. PMID- 15117929 TI - Severe bilateral optic neuritis associated with prolonged linezolid therapy. PMID- 15117930 TI - Effects of triclosan incorporation into ABS plastic on biofilm communities. AB - OBJECTIVE: This study compared the attached biofilm populations on acrylonitrile butadiene-styrene (ABS) plastic with and without the incorporation of the antimicrobial triclosan [5-chloro-2-(2,4-dichlorophenoxy) phenol] after 1-3 weeks of exposure to drinking water. METHODS: Biofilms were cultivated on triclosan incorporated (TP) and control plastics (CP) in continuous flow culture reactors with drinking water as the growth medium and inoculum. After 1-3 weeks of exposure, the plastics were removed and the biofilms aseptically harvested. The attached communities were examined with respect to direct cell counts, culturability, triclosan resistance and community composition. RESULTS: Based on these analyses, no significant differences were observed between the populations attached to TP and CP surfaces. Results from both a bioavailability assay and gas chromatography mass spectrometry analyses, revealed that only trace amounts of triclosan desorbed from the plastic. The lack of biofilm community difference, coupled with this limited desorption of triclosan from the TP indicates that the ABS plastic studied was no more effective at controlling bacterial populations than the control plastic because the antimicrobial was not bioavailable. CONCLUSIONS: These results call into question the long-term utility of triclosan incorporation into ABS plastic and highlight the need for proof of efficacy regarding the antimicrobial properties of such materials. PMID- 15117931 TI - Class 1 integrons in various Salmonella enterica serovars isolated from animals and identification of genomic island SGI1 in Salmonella enterica var. Meleagridis. AB - OBJECTIVES: To determine the prevalence of integron-mediated antibiotic resistance in a diverse sample set of Salmonella enterica isolated from animals. MATERIALS AND METHODS: Multiplex PCR was used to detect class 1 integron gene sequences, and integron gene cassettes were identified by PCR mapping. Susceptibility to 18 antibiotics or antibiotic combinations commonly used in either human or veterinary medicine was measured using a microdilution method, and statistical comparisons of the frequency of resistance between groups were made using Fisher's two-sided probability test. Genotypic comparisons of isolates were made following pulsed-field gel electrophoresis of genomic DNA. RESULTS: Thirty-two (30.8%) of 104 isolates contained class 1 integron sequences. Integron positive isolates represented 15 different S. enterica serovars, were obtained from nine different animal species and had a higher frequency of non-integron mediated antibiotic resistance (P < 0.05) compared with integron-negative isolates. One non-Typhimurium isolate (S. enterica Meleagridis) contained an SGI1 genomic island, including the antibiotic resistance gene cluster. CONCLUSIONS: These data demonstrate that integron-mediated antibiotic resistance is common among diverse Salmonella serovars, many of them rare. In addition, SGI1 is not limited to Salmonella enterica Typhimurium DT104 or other commonly isolated serovars. PMID- 15117932 TI - Antibiotic treatment of gram-positive bone and joint infections. AB - Gram-positive organisms, particularly staphylococci and streptococci, are responsible for the majority of bone and joint infections. Treatment of these infections can be difficult, usually involving a prolonged course of antibiotics, often with surgical intervention. The selection of antibiotics depends on sensitivity profile, patient tolerance and long-term goals, e.g. cure or suppression, but there are few randomized controlled trials in patients comparing efficacy of different antibiotics. Different degrees of bone penetration and clinical outcome for specific antibiotics, e.g. the beta-lactams, clindamycin and quinolones, have been described, although the methodology in these studies is not standardized and findings cannot always be applied directly to patients. The effect of attaining minimum serum bactericidal concentrations in patients has also been studied but this is no longer routinely recommended in clinical practice. Comparative clinical trials are few but have demonstrated efficacy of oral fluoroquinolones in combination with either rifampicin or fusidic acid for selected Gram-positive infections. In the past decade, increasingly resistant organisms, e.g. methicillin-resistant Staphylococcus aureus and vancomycin resistant enterococci have been recognized as causes of orthopaedic infection. Individual case reports describe successful treatment using the newer antibiotics, e.g. linezolid and quinupristin/dalfopristin, but results of clinical trials are awaited. PMID- 15117933 TI - Towards targeted prescribing: will the cure for antimicrobial resistance be specific, directed therapy through improved diagnostic testing? AB - The discovery of antimicrobial agents was one of the major events of the twentieth century. However, with the 'antibiotic era' barely five decades old, we are now faced with the global problem of emerging resistance in virtually all pathogens. Guidelines and admonishments to improve prescribing have had little effect. At this point, in the twenty-first century, we are on the threshold of another era of discovery-that of molecular diagnostics. We postulate that the development and use of new molecular microbiological testing, coupled with an ever-improving understanding of how best to use these precious drugs in the treatment of infection, offers the greatest hope yet for physician prescribing that can retard, or perhaps even reduce, the development of drug resistance in many microbial species. This diagnostic advance could preserve the utility of antimicrobial agents well into the future for the benefit of all people. PMID- 15117934 TI - Clinical efficacy of ketolides in the treatment of respiratory tract infections. AB - Ketolides are a new class of semi-synthetic agents derived from erythromycin A designed to overcome erythromycin A resistance in Streptococcus pneumoniae. Telithromycin (HMR 3647) is the first member of this new class to be approved for clinical use. Cethromycin (ABT-773) has been developed up to Phase III, but its further development seems questionable at the moment. Other ketolides are only in the first stages of preclinical development and may not be available within the foreseeable future. Ketolide compounds inhibit bacterial protein synthesis by interacting with the peptidyl transferase site of the 50S ribosomal subunit, and interact closely with domains II at A752 and V at A2058 and A2059 of the 23S rRNA. These compounds also inhibit the formation of the 50S subunit of the ribosome. Ketolides show good activity against the Gram-positive bacteria responsible for respiratory tract infections including penicillin G- and erythromycin A-resistant S. pneumoniae. The 15 clinical trials with telithromycin published to date include four randomized, double-blind comparative trials and three open-label studies in community-acquired pneumonia, three randomized double blind trials in acute exacerbation of chronic bronchitis, two randomized double blind trials in pharyngitis, and two double-blind comparative trials and one open label trial in acute maxillary sinusitis. Clinical response rates were favourable in all clinical trials, with eradication rates in patients with pneumococcal bacteraemia and penicillin G- and erythromycin A-resistant pneumococcal infections at least as high as those of comparators. As resistance to macrolides continues to emerge, the availability of other ketolides besides telithromycin and a development programme for the application of ketolides in children would appear to be warranted to obtain a new class of antibiotics that may one day replace macrolides. PMID- 15117935 TI - Selection for high-level resistance by chronic triclosan exposure is not universal. AB - OBJECTIVES: To investigate the effect of triclosan exposure on the antimicrobial susceptibilities of numerically important dental bacteria. METHODS: A gradient plate technique was used to expose Fusobacterium nucleatum, Lactobacillus rhamnosus, Neisseria subflava, Porphyromonas gingivalis, Actinomyces naeslundii, Prevotella nigrescens, Streptococcus oralis, Streptococcus sanguis, Streptococcus mutans and Veillonella dispar repeatedly to escalating, sublethal concentrations of triclosan. Escherichia coli ATCC 8739 was included as an organism showing the triclosan resistance development trait. MIC values towards chlorhexidine, metronidazole and tetracycline were determined before and after biocide exposure. RESULTS: N. subflava, Pr. nigrescens Po. gingivalis and E. coli were highly susceptible to triclosan (MIC range 0.1-3.9 mg/L), whereas the lactobacillus and S. mutans were less susceptible (MIC range 15.6-20.8 mg/L). Triclosan exposure resulted in a highly significant ( approximately 400-fold) reduction in triclosan susceptibility (P < 0.01) for the positive control E. coli, although its MICs towards chlorhexidine, metronidazole and tetracycline were not significantly altered. Minor ( approximately two-fold) decreases in triclosan susceptibility (MIC) occurred for Pr. nigrescens and in S. sanguis and S. oralis (MBC). Mean changes in susceptibilities (MIC and MBC) of the oral species to chlorhexidine, metronidazole and tetracycline did not exceed two-fold, although chlorhexidine MBCs for S. sanguis were markedly, but transiently, increased. CONCLUSIONS: These data fail to demonstrate biologically significant drug resistance in triclosan exposed bacteria and suggest that markedly decreased triclosan susceptibility, although confirmed for E. coli, is not a universal phenomenon. Other bacteria possibly possess more susceptible targets than FabI that are highly conserved, which may govern triclosan activity. PMID- 15117936 TI - Cloning and characterization of a novel mannose-binding protein of Acanthamoeba. AB - Acanthamoebae produce a painful, blinding infection of the cornea. The mannose binding protein (MBP) of Acanthamoeba is thought to play a key role in the pathogenesis of the infection by mediating the adhesion of parasites to the host cells. We describe here the isolation and molecular cloning of Acanthamoeba MBP. The MBP was isolated by chromatography on the mannose affinity gel. Gel filtration experiments revealed that the Acanthamoeba lectin is a approximately 400-kDa protein that is constituted of multiple 130-kDa subunits. Cloning and sequencing experiments indicated that the Acanthamoeba MBP gene is composed of 6 exons and 5 introns that span 3.6 kb of the amoeba genome and that MBP cDNA codes for a precursor protein of 833 amino acids. That the cloned cDNA encodes authentic MBP was demonstrated by showing that: (i). recombinant MBP possesses mannose binding activity, and (ii). polyclonal antibodies prepared against Acanthamoeba MBP bound to the recombinant protein. Sequence analysis revealed that the MBP contains a large N-terminal extracellular domain, a transmembrane domain, and a short C-terminal cytoplasmic domain. Despite extensive BLAST searches using the MBP sequence, no significant matches were retrieved. The most striking feature of the Acanthamoeba MBP sequence is the presence of a cysteine rich region containing 14 CXCXC motifs within the extracellular domain. In summary, we have isolated, cloned, and characterized a novel MBP from Acanthamoeba. Because the presence of antibodies to MBP in tears provides protection against infection, the availability of the MBP cDNA sequence and rMBP should help develop: (i). a tear-based test to identify individuals who are at risk of developing the keratitis and (ii). strategies to immunize high-risk individuals. PMID- 15117937 TI - Identification and activity of a series of azole-based compounds with lactate dehydrogenase-directed anti-malarial activity. AB - Plasmodium falciparum, the causative agent of malaria, relies extensively on glycolysis coupled with homolactic fermentation during its blood-borne stages for energy production. Selective inhibitors of the parasite lactate dehydrogenase (LDH), central to NAD(+) regeneration, therefore potentially provide a route to new antimalarial drugs directed against a novel molecular target. A series of heterocyclic, azole-based compounds are described that preferentially inhibit P. falciparum LDH at sub-micromolar concentrations, typically at concentrations about 100-fold lower than required for human lactate dehydrogenase inhibition. Crystal structures show these competitive inhibitors form a network of interactions with amino acids within the active site of the enzyme, stacking alongside the nicotinamide ring of the NAD(+) cofactor. These compounds display modest activity against parasitized erythrocytes, including parasite strains with known resistance to existing anti-malarials and against Plasmodium berghei in BALB/c mice. Initial toxicity data suggest the azole derivatives have generally low cytotoxicity, and preliminary pharmoco-kinetic data show favorable bioavailability and circulation times. These encouraging results suggest that further enhancement of these structures may yield candidates suitable for consideration as new therapeutics for the treatment of malaria. In combination these studies also provide strong support for the validity of targeting the Plasmodium glycolytic pathway and, in particular, LDH in the search for novel anti-malarials. PMID- 15117938 TI - RNA sequence elements required for high affinity binding by the zinc finger domain of tristetraprolin: conformational changes coupled to the bipartite nature of Au-rich MRNA-destabilizing motifs. AB - Tristetraprolin (TTP) binds AU-rich elements (AREs) encoded within selected labile mRNAs and targets these transcripts for rapid cytoplasmic decay. RNA binding by TTP is mediated by an approximately 70-amino acid domain containing two tandemly arrayed CCCH zinc fingers. Here we show that a 73-amino acid peptide spanning the TTP zinc finger domain, denoted TTP73, forms a dynamic, equimolar RNA.peptide complex with a 13-nucleotide fragment of the ARE from tumor necrosis factor alpha mRNA, which includes small but significant contributions from ionic interactions. Association of TTP73 with high affinity RNA substrates is accompanied by a large negative change in heat capacity without substantial modification of RNA structure, consistent with conformational changes in the peptide moiety during RNA binding. Analyses using mutant ARE substrates indicate that two adenylate residues located 3-6 bases apart within a uridylate-rich sequence are sufficient for high affinity recognition by TTP73 (K(d) <20 nm), with optimal affinity observed for RNA substrates containing AUUUA or AUUUUA. Linkage of conformational changes and binding affinity to the presence and spacing of these adenylate residues provides a thermodynamic basis for the RNA substrate specificity of TTP. PMID- 15117939 TI - The X-ray structure of human mannan-binding lectin-associated protein 19 (MAp19) and its interaction site with mannan-binding lectin and L-ficolin. AB - MAp19 is an alternative splicing product of the MASP-2 gene comprising the N terminal CUB1-epidermal growth factor (EGF) segment of MASP-2, plus four additional residues at its C-terminal end. Like full-length MASP-2, it forms Ca(2+)-dependent complexes with mannan-binding lectin (MBL) and L-ficolin. The x ray structure of human MAp19 was solved to a resolution of 2.5 A. It shows a head to tail homodimer held together by interactions between the CUB1 module of one monomer and the EGF module of its counterpart. A Ca(2+) ion bound to each EGF module stabilizes the dimer interfaces. A second Ca(2+) ion is bound to the distal end of each CUB1 module, through six ligands contributed by Glu(52), Asp(60), Asp(105), Ser(107), Asn(108), and a water molecule. Compared with its counterpart in human C1s, the N-terminal end of the MAp19 CUB1 module contains a 7-residue extension that forms additional inter-monomer contacts. To identify the residues involved in the interaction of MAp19 with MBL and L-ficolin, point mutants were generated and their binding ability was determined using surface plasmon resonance spectroscopy. Six mutations at Tyr(59), Asp(60), Glu(83), Asp(105), Tyr(106), and Glu(109) either strongly decreased or abolished interaction with both MBL and L-ficolin. These mutations map a common binding site for these proteins located at the distal end of each CUB1 module and stabilized by the Ca(2+) ion. PMID- 15117940 TI - Clathrin-mediated endocytosis of m3 muscarinic receptors. Roles for Gbetagamma and tubulin. AB - Receptors as well as some G protein subunits internalize after agonist stimulation. It is not clear whether Galpha(q) or Gbetagamma undergo such regulated translocation. Recent studies demonstrate that m3 muscarinic receptor activation in SK-N-SH neuroblastoma cells causes recruitment of tubulin to the plasma membrane. This subsequently transactivates Galpha(q) and activates phospholipase Cbeta1. Interaction of tubulin-GDP with Gbetagamma at the offset of phospholipase Cbeta1 signaling appears involved in translocation of tubulin and Gbetagamma to vesicle-like structures in the cytosol (Popova, J. S., and Rasenick, M. M. (2003) J. Biol. Chem. 278, 34299-34308). The relationship of this internalization to the clathrin-mediated endocytosis of the activated m3 muscarinic receptors or Galpha(q) involvement in this process has not been clarified. To test this, SK-N-SH cells were treated with carbachol, and localization of Galpha(q), Gbetagamma, tubulin, clathrin, and m3 receptors were analyzed by both cellular imaging and biochemical techniques. Upon agonist stimulation both tubulin and clathrin translocated to the plasma membrane and co localized with receptors, Galpha(q) and Gbetagamma. Fifteen minutes later receptors, Gbetagamma and tubulin, but not Galpha(q), internalized with the clathrin-coated vesicles. Coimmunoprecipitation of m3 receptors with Gbetagamma, tubulin, and clathrin from the cytosol of carbachol-treated cells was readily observed. These data suggested that Gbetagamma subunits might organize the formation of a multiprotein complex linking m3 receptors to tubulin since they interacted with both proteins. Such protein assemblies might explain the dynamin dependent but beta-arrestin-independent endocytosis of m3 muscarinic receptors since tubulin interaction with dynamin might guide or insert the complex into clathrin-coated pits. This novel mechanism of internalization might prove important for other beta-arrestin-independent endocytic pathways. It also suggests cross-regulation between G protein-mediated signaling and the dynamics of the microtubule cytoskeleton. PMID- 15117941 TI - Subfertility, uterine hypoplasia, and partial progesterone resistance in mice lacking the Kruppel-like factor 9/basic transcription element-binding protein-1 (Bteb1) gene. AB - Progesterone receptor (PR), a ligand-activated transcription factor, is a key regulator of cellular proliferation and differentiation in reproductive tissues. The transcriptional activity of PR is influenced by co-regulatory proteins typically expressed in a tissue- and cell-specific fashion. We previously demonstrated that basic transcription element-binding protein-1 (BTEB1), a member of the Sp/Kruppel-like family of transcription factors, functionally interacts with the two PR isoforms, PR-A and PR-B, to mediate progestin sensitivity of target genes in endometrial epithelial cells in vitro. Here we report that ablation of the Bteb1 gene in female mice results in uterine hypoplasia, reduced litter size, and increased incidence of neonatal deaths in offspring. The reduced litter size is solely a maternal genotype effect and results from fewer numbers of implantation sites, rather than defects in ovulation. In the early pregnant uterus, Bteb1 expression in stromal cells temporally coincides with PR-A isoform dependent decidual formation at the time of implantation. Expression of two implantation-specific genes, Hoxa10 and cyclin D3, was decreased in uteri of early pregnant Bteb1-null mutants, whereas that of Bteb3, a related family member, was increased, the latter possibly compensating for the loss of Bteb1. Progesterone responsiveness of several uterine genes was altered with Bteb1-null mutation. These results identify Bteb1 as a functionally relevant PR-interacting protein and suggest its selective modulation of cellular processes that are regulated by PR-A in the uterine stroma. PMID- 15117942 TI - Dual role of sumoylation in the nuclear localization and transcriptional activation of NFAT1. AB - The nuclear import of nuclear factor of activated T cells (NFAT) transcription factors is critical for regulating NFAT activity. Here we demonstrate that the sumoylation of NFAT1 defines a novel mechanism of the nuclear anchorage and transcriptional activation downstream from the known mechanism of calcineurin mediated dephosphorylation and nuclear import. We show that Lys(684) and Lys(897) of NFAT1 can be sumoylated. The sumoylation at Lys(684) is required for NFAT1 transcriptional activity and subsequent sumoylation of Lys(897), whereas the sumoylation of Lys(897) is only required for nuclear anchorage. Because Lys(897) of NFAT1 is not conserved among other members of the NFAT family, we propose that sumoylation of Lys(897) may provide a mechanism for NFAT1 isotype-specific regulation of nuclear anchorage and transcriptional activation. Furthermore, we found that treatment with both ionomycin and phorbol 12-myristate 13-acetate ensured efficient nuclear anchorage with the recruitment of NFAT1 into the SUMO-1 bodies, whereas treatment with ionomycin alone induced nuclear translocation of NFAT1 but not recruitment into the SUMO-1 bodies. Our results suggest that the recruitment of NFAT1 into SUMO-1 bodies may be required for the progressive transcriptional activity of NFAT1 upon co-stimulation with ionomycin and phorbol 12-myristate 13-acetate, whereas anergic transcription stimulated by ionomycin alone may occur without recruitment into the SUMO-1 bodies. PMID- 15117943 TI - Sem1p is a novel subunit of the 26 S proteasome from Saccharomyces cerevisiae. AB - The 26 S proteasome, which catalyzes degradation of polyubiquitinated proteins, is composed of the 20 S proteasome and the 19 S regulatory particle (RP). The RP is composed of the lid and base subcomplexes and regulates the catalytic activity of the 20 S proteasome. In this study, we carried out affinity purification of the lid and base subcomplexes from the tagged strains of Saccharomyces cerevisiae, and we found that the lid contains a small molecular mass protein, Sem1. The Sem1 protein binds with the 26 S proteasome isolated from a mutant with deletion of SEM1 but not with the 26 S proteasome from the wild type. The lid lacking Sem1 is unstable at a high salt concentration. The 19 S RP was immunoprecipitated together with Sem1 by immunoprecipitation using hemagglutinin epitope-tagged Sem1 as bait. Degradation of polyubiquitinated proteins in vivo or in vitro is impaired in the Sem1-deficient 26 S proteasome. In addition, genetic interaction between SEM1 and RPN10 was detected. The human Sem1 homologue hDSS1 was found to be a functional homologue of Sem1 and capable of interacting with the human 26 S proteasome. The results suggest that Sem1, possibly hDSS1, is a novel subunit of the 26 S proteasome and plays a role in ubiquitin-dependent proteolysis. PMID- 15117944 TI - Phosphorylation of alphaB-crystallin alters chaperone function through loss of dimeric substructure. AB - Phosphorylation is the most common posttranslational modification of the alpha crystallins in the human lens. These phosphorylated forms are not only important because of their abundance in aging lenses and the implications for cataract but also because they have been identified in patients with degenerative brain disease. By using mimics corresponding to the reported in vivo phosphorylation sites in the human lens, we have examined the effects of phosphorylation upon the chaperone-like properties and structure of alphaB-crystallin. Here we show that phosphorylation of alphaB-crystallin at Ser-45 results in uncontrolled aggregation. By using an innovative tandem mass spectrometry approach, we demonstrate how this alteration in behavior stems from disruption of dimeric substructure within the polydisperse alphaB-crystallin assembly. This structural perturbation appears to disturb the housekeeping role of alphaB-crystallin and consequently has important implications for the disease states caused by protein aggregation in the lens and deposition in non-lenticular tissue. PMID- 15117945 TI - The Human hydroxyacylglutathione hydrolase (HAGH) gene encodes both cytosolic and mitochondrial forms of glyoxalase II. AB - In yeast and higher plants, separate genes encode the cytosolic and mitochondrial forms of glyoxalase II. In contrast, although glyoxalase II activity has been detected both in the cytosol and mitochondria of mammals, only a single gene encoding glyoxalase II has been identified. Previously it was thought that this gene (the hydroxyacylglutathione hydrolase gene), comprised 8 exons that are transcribed into mRNA and that the resulting mRNA species encoded a single cytosolic form of glyoxalase II. Here we show that this gene gives rise to two distinct mRNA species transcribed from 9 and 10 exons, respectively. The 9-exon derived transcript encodes two protein species: mitochondrially targeted glyoxylase II, which is initiated from an AUG codon in a previously uncharacterized part of the mRNA sequence, and cytosolic glyoxalase II, which is initiated by internal ribosome entry at a downstream AUG codon. The transcript deriving from 10 exons has an in-frame termination codon between the two initiating AUG codons and hence only encodes the cytosolic form of the protein. Confocal fluorescence microscopy indicates that the mitochondrially targeted form of glyoxalase II is directed to the mitochondrial matrix. Analysis of glyoxalase II mRNA sequences from a number of species indicates that dual initiation from alternative AUG codons is conserved throughout vertebrates. PMID- 15117946 TI - Maltose-binding protein is open in the catalytic transition state for ATP hydrolysis during maltose transport. AB - The maltose transport complex of Escherichia coli, a member of the ATP-binding cassette superfamily, mediates the high affinity uptake of maltose at the expense of ATP. The membrane-associated transporter consists of two transmembrane subunits, MalF and MalG, and two copies of the cytoplasmic ATP-binding cassette subunit, MalK. Maltose-binding protein (MBP), a soluble periplasmic protein, delivers maltose to the MalFGK(2) transporter and stimulates hydrolysis by the transporter. Site-directed spin labeling electron paramagnetic resonance spectroscopy is used to monitor binding of MBP to MalFGK(2) and conformational changes in MBP as it interacts with MalFGK(2). Cysteine residues and spin labels have been introduced into the two lobes of MBP so that spin-spin interaction will report on ligand-induced closure of the protein (Hall, J. A., Thorgeirsson, T. E., Liu, J., Shin, Y. K., and Nikaido, H. (1997) J. Biol. Chem. 272, 17610 17614). At least two different modes of interaction between MBP and MalFGK(2) were detected. Binding of MBP to MalFGK(2) in the absence of ATP resulted in a decrease in motion of spin label at position 41 in the C-terminal domain of MBP. In a vanadate-trapped transition state intermediate, all free MBP became tightly bound to MalFGK(2), spin label in both lobes became completely immobilized, and spin-spin interactions were lost, suggesting that MBP was in an open conformation. Binding of non-hydrolyzable MgATP analogs or ATP in the absence of Mg is sufficient to stabilize a complex of open MBP and MalFGK(2). Taken together, these data suggest that closure of the MalK dimer interface coincides with opening of MBP and maltose release to the transporter. PMID- 15117947 TI - Acrylodan-conjugated cysteine side chains reveal conformational state and ligand site locations of the acetylcholine-binding protein. AB - We undertook cysteine substitution mutagenesis and fluorophore conjugation at selected residue positions to map sites of ligand binding and changes in solvent exposure of the acetylcholine-binding protein from Lymnaea stagnalis, a nicotinic receptor surrogate. Acrylodan fluorescence emission is highly sensitive to its local environment, and when bound to protein, exhibits changes in both intensity and emission wavelength that are reflected in the degree of solvent exclusion and the effective dielectric constant of the environment of the fluorophore. Hence, cysteine mutants were generated based on the acetylcholine-binding protein crystal structure and predicted ligand binding sites, and fluorescence parameters were assayed on the acrylodan-conjugated proteins. This approach allows one to analyze the environment around the conjugated fluorophore side chain and the changes induced by bound ligand. Introduction of an acrylodan-cysteine conjugate at position 178 yields a large blue shift with alpha-bungarotoxin association, whereas the agonists and alkaloid antagonists induce red shifts reflecting solvent exposure at this position. Such residue-selective changes in fluorescence parameters suggest that certain ligands can induce distinct conformational states of the binding protein, and that mutually exclusive binding results from disparate portals of entry to and orientations of the bound alpha-toxin and smaller acetylcholine congeners at the binding pocket. Labeling at other residue positions around the predicted binding pocket also reveals distinctive spectral changes for alpha-bungarotoxin, agonists, and alkaloid antagonists. PMID- 15117948 TI - The SANT domain of human MI-ER1 interacts with Sp1 to interfere with GC box recognition and repress transcription from its own promoter. AB - To gain insight into the regulation of hmi-er1 expression, we cloned a human genomic DNA fragment containing one of the two hmi-er1 promoters and consisting of 1460 bp upstream of the translation initiation codon of hMI-ER1. Computer assisted sequence analysis revealed that the hmi-er1 promoter region contains a CpG island but lacks an identifiable TATA element, initiator sequence and downstream promoter element. This genomic DNA was able to direct transcription of a luciferase reporter gene in a variety of human cell lines, and the minimal promoter was shown to be located within-68/+144 bp. Several putative Sp1 binding sites were identified, and we show that Sp1 can bind to the hmi-er1 minimal promoter and increase transcription, suggesting that the level of hmi-er1 expression may depend on the availability of Sp1 protein. Functional analysis revealed that hMI-ER1 represses Sp1-activated transcription from the minimal promoter by a histone deacetylase-independent mechanism. Chromatin immunoprecipitation analysis demonstrated that both Sp1 and hMI-ER1 are associated with the chromatin of the hmi-er1 promoter and that overexpression of hMI-ER1 in cell lines that allow Tet-On-inducible expression resulted in loss of detectable Sp1 from the endogenous hmi-er1 promoter. The mechanism by which this occurs does not involve binding of hMI-ER1 to cis-acting elements. Instead, we show that hMI-ER1 physically associates with Sp1 and that endogenous complexes containing the two proteins could be detected in vivo. Furthermore, hMI-ER1 specifically interferes with binding of Sp1 to the hmi-er1 minimal promoter as well as to an Sp1 consensus oligonucleotide. Deletion analysis revealed that this interaction occurs through a region containing the SANT domain of hMI-ER1. Together, these data reveal a functional role for the SANT domain in the action of co-repressor regulatory factors and suggest that the association of hMI-ER1 with Sp1 represents a novel mechanism for the negative regulation of Sp1 target promoters. PMID- 15117949 TI - Rad23 and Rpn10 serve as alternative ubiquitin receptors for the proteasome. AB - The selective recognition of ubiquitin conjugates by proteasomes is a key step in protein degradation. The receptors that mediate this step have yet to be clearly defined although specific candidates exist. Here we show that the proteasome directly recognizes ubiquitin chains through a specific subunit, Rpn10, and also recognizes chains indirectly through Rad23, a reversibly bound proteasome cofactor. Both binding events can be observed in purified biochemical systems. A block substitution in the chain-binding ubiquitin interacting motif of RPN10 when combined with a null mutation in RAD23 results in a synthetic defect in protein degradation consistent with the view that the direct and indirect recognition modes function to some extent redundantly in vivo. Rad23 and the deubiquitinating enzyme Ubp6 both bind proteasome subunit Rpn1 through N-terminal ubiquitin-like domains. Surprisingly, Rad23 and Ubp6 do not compete with each other for proteasome binding. Thus, Rpn1 may act as a scaffold to assemble on the proteasome multiple proteins that act to either bind or hydrolyze multiubiquitin chains. PMID- 15117950 TI - Regulation of ubiquitin protein ligase activity in c-Cbl by phosphorylation induced conformational change and constitutive activation by tyrosine to glutamate point mutations. AB - c-Cbl down-regulates receptor tyrosine kinases by conjugating ubiquitin to them, leading to receptor internalization and degradation. The ubiquitin protein ligase activity of c-Cbl (abbreviated as E3 activity) is mediated by its RING finger domain. We show here that the E3 activity of c-Cbl is negatively regulated by other domains present in the amino-terminal half of the protein (the TKB and linker helix domains) and that this negative regulation is removed when the protein is phosphorylated on tyrosine residues. Protease digestion studies indicate that tyrosine phosphorylation alters the conformation of c-Cbl. We also show that mutation of certain conserved tyrosine residues to glutamate can constitutively activate the E3 activity of c-Cbl. In particular, a Y371E mutant shows constitutive E3 activity while retaining the ability to bind epidermal growth factor receptor (EGFR). The Y371E mutant also has altered protease sensitivity from wild type, instead resembling the proteolytic pattern seen with tyrosine-phosphorylated c-Cbl. Mutation of the homologous tyrosine residue in Cbl b to glutamate also leads to E3 activation while retaining EGFR-binding ability. These studies argue that Tyr-371 plays a key role in activating the E3 activity of c-Cbl and that the Y371E mutant may partially mimic phosphorylation at that site. However, Tyr-371 point mutants of c-Cbl are still able to undergo phosphorylation-induced E3 activation, and we show that Tyr-368 can also be phosphorylated in addition to Tyr-371, and contributes to activation. PMID- 15117951 TI - The insect hemolymph protein HP19 mediates the nongenomic effect of ecdysteroids on acid phosphatase activity. AB - The activity of acid phosphatase (ACP) in insect fat bodies is stimulated by the steroid hormone 20-hydoxyecdysone (20E) in vivo. However, in fat bodies kept in culture, a factor from the hemolymph is required to enhance the ACP activity. We identified the factor as a protein with a molecular mass of 19 kDa (HP19) from the hemolymph of a lepidopteran insect, the rice moth, Corcyra cephalonica. Western analysis of hemolymph proteins with denaturing and non-denaturing PAGE using antibodies raised against HP19 suggest that this protein exists as a monomer. It is synthesized by the hind gut-associated lobular fat body of the larvae and is released into the hemolymph. The stimulatory effect of HP19 on the ACP activity is developmentally regulated and exhibits its maximal effect shortly before the onset of metamorphosis. We cloned the HP19 cDNA by immunoscreening a hind gut-associated lobular fat body cDNA expression library. Analysis of the amino acid sequence shows that HP19 belongs to the family of glutathione S transferase (GST) like proteins. However, affinity-purified GST from Corcyra failed to show any mediation effect on 20E-stimulated ACP activity, and HP19 lacks GST enzymatic activity. Notably, HP19 mediates the hormone-stimulated ACP activity in intact fat body tissue and homogenates even in the presence of inhibitors of transcription and translation, suggesting a nongenomic mode of action. In addition, we show that HP19 inhibits the 20E-induced phosphorylation of the hexamerin receptor protein. PMID- 15117952 TI - Crystal structure of hormone-bound atrial natriuretic peptide receptor extracellular domain: rotation mechanism for transmembrane signal transduction. AB - A cardiac hormone, atrial natriuretic peptide (ANP), plays a major role in blood pressure and volume regulation. ANP activities are mediated by a single span transmembrane receptor carrying intrinsic guanylate cyclase activity. ANP binding to its extracellular domain stimulates guanylate cyclase activity by an as yet unknown mechanism. Here we report the crystal structure of dimerized extracellular hormone-binding domain in complex with ANP. The structural comparison with the unliganded receptor reveals that hormone binding causes the two receptor monomers to undergo an intermolecular twist with little intramolecular conformational change. This motion produces a Ferris wheel-like translocation of two juxtamembrane domains in the dimer with essentially no change in the interdomain distance. This movement alters the relative orientation of the two domains by a shift equivalent to counterclockwise rotation of each by 24 degrees. These results suggest that transmembrane signaling by the ANP receptor is initiated via a hormone-induced rotation mechanism. PMID- 15117953 TI - Requirement for aspartate-cleaved bid in apoptosis signaling by DNA-damaging anti cancer regimens. AB - Lymphoid malignancies can escape from DNA-damaging anti-cancer drugs and gamma radiation by blocking apoptosis-signaling pathways. How these regimens induce apoptosis is incompletely defined, especially in cells with nonfunctional p53. We report here that the BH3-only Bcl-2 family member Bid is required for mitochondrial permeabilization and apoptosis induction by etoposide and gamma radiation in p53 mutant T leukemic cells. Bid is not transcriptionally up regulated in response to these stimuli but is activated by cleavage on aspartate residues 60 and/or 75, which are the targets of caspase-8 and granzyme B. Bid activity is not inhibitable by c-Flip(L), CrmA, or dominant negative caspase-9 and therefore is independent of inducer caspase activation by death receptors or the mitochondria. Caspase-2, which has been implicated as inducer caspase in DNA damage pathways, appeared to be processed in response to etoposide and gamma radiation but downstream of caspase-9. Knock down of caspase-2 by short interfering RNA further excluded its role in Bid activation by DNA damage. Caspase-2 was implicated in the death receptor pathway however, where it contributed to effector caspase processing downstream of inducer caspases. Granzyme B-specific serpins could not block DNA damage-induced apoptosis, excluding a role for granzyme B in the generation of active Bid. We conclude that Bid, cleaved by an undefined aspartate-specific protease, can be a key mediator of the apoptotic response to DNA-damaging anticancer regimens. PMID- 15117954 TI - Asr1p, a novel yeast ring/PHD finger protein, signals alcohol stress to the nucleus. AB - During fermentation, yeast cells are exposed to increasing amounts of alcohol, which is stressful and affects both growth and viability. On the molecular level, numerous aspects of alcohol stress signaling remain unresolved. We have identified a novel yeast Ring/PHD finger protein that constitutively shuttles between nucleus and cytoplasm but accumulates in the nucleus upon exposure to ethanol, 2-propanol, or 1-butanol. Subcellular localization of this protein is not altered by osmotic, oxidative, or heat stress or during nitrogen or glucose starvation. Because of its exclusive sensitivity to environmental alcohol, the protein was called Asr1p for Alcohol Sensitive Ring/PHD finger 1 protein. Nuclear accumulation of Asr1p is rapid, reversible, and requires a functional Ran/Gsp1p gradient. Asr1p contains two N terminally located leucine-rich nuclear export sequences (NES) required for nuclear export. Consistently, it accumulates in the nucleus of xpo1-1 cells at restrictive temperature and forms a trimeric complex with the exportin Xpo1p and Ran-GTP. Deletion of ASR1 leads to sensitivity in growth on medium containing alcohol or detergent, consistent with a function of Asr1p in alcohol-related signaling. Asr1p is the first reported protein that changes its subcellular localization specifically upon exposure to alcohol and therefore represents a key element in the analysis of alcohol-responsive signaling. PMID- 15117955 TI - Novel mechanisms of pH sensitivity in tuna hemoglobin: a structural explanation of the root effect. AB - The crystal structure of hemoglobin has been known for several decades, yet various features of the molecule remain unexplained or controversial. Several animal hemoglobins have properties that cannot be readily explained in terms of their amino acid sequence and known atomic models of hemoglobin. Among these, fish hemoglobins are well known for their widely varying interactions with heterotropic effector molecules and pH sensitivity. Some fish hemoglobins are almost completely insensitive to pH (within physiological limits), whereas others show extremely low oxygen affinity under acid conditions, a phenomenon called the Root effect. X-ray crystal structures of Root effect hemoglobins have not, to date, provided convincing explanations of this effect. Sequence alignments have signally failed to pinpoint the residues involved, and site-directed mutagenesis has not yielded a human hemoglobin variant with this property. We have solved the crystal structure of tuna hemoglobin in the deoxy form at low and moderate pH and in the presence of carbon monoxide at high pH. A comparison of these models shows clear evidence for novel mechanisms of pH-dependent control of ligand affinity. PMID- 15117956 TI - Transcriptional mechanisms regulating alveolar epithelial cell-specific CCL5 secretion in pulmonary tuberculosis. AB - CCL5 (or RANTES (regulated upon activation, normal T cell expressed and secreted)) recruits T lymphocytes and monocytes. The source and regulation of CCL5 in pulmonary tuberculosis are unclear. Infection of the human alveolar epithelial cell line (A549) by Mycobacterium tuberculosis caused no CCL5 secretion and little monocyte secretion. Conditioned medium from tuberculosis infected human monocytes (CoMTB) stimulated significant CCL5 secretion from A549 cells and from primary alveolar, but not upper airway, epithelial cells. Differential responsiveness of small airway and normal human bronchial epithelial cells to CoMTB but not to conditioned medium from unstimulated human monocytes was specific to CCL5 and not to CXCL8. CoMTB induced CCL5 mRNA accumulation in A549 cells and induced nuclear translocation of nuclear factor kappaB (NFkappaB) subunits p50, p65, and c-rel at 1 h; nuclear binding of activator protein (AP)-1 (c-Fos, FosB, and c-Jun) at 4-8 h; and binding of NF-interleukin (IL)-6 at 24 h. CCL5 promoter-reporter analysis using deletion and site-specific mutagenesis constructs demonstrated a key role for AP-1, NF-IL-6, and NFkappaB in driving CoMTB-induced promoter activity. The IL-1 receptor antagonist inhibited A549 and small airway epithelial cell CCL5 secretion, gene expression, and promoter activity. CoMTB contained IL-1beta, and recombinant IL-1beta reproduced CoMTB effects. Monocyte alveolar, but not upper airway, epithelial cell networks in pulmonary tuberculosis cause AP-1-, NF-IL-6-, and NFkappaB-dependent CCL5 secretion. IL-1beta is the critical regulator of tuberculosis-stimulated CCL5 secretion in the lung. PMID- 15117957 TI - Crystal structure of the membrane fusion protein, MexA, of the multidrug transporter in Pseudomonas aeruginosa. AB - The MexAB-OprM efflux pump of Pseudomonas aeruginosa is central to multidrug resistance of this organism, which infects immunocompromised hospital patients. The MexA, MexB, and OprM subunits were assumed to function as the membrane fusion protein, the body of the transporter, and the outer membrane channel protein, respectively. For better understanding of this important xenobiotic transporter, we show the x-ray crystallographic structure of MexA at a resolution of 2.40 A. The global MexA structure showed unforeseen new features with a spiral assembly of six and seven protomers that were joined together at one end by a pseudo 2 fold image. The protomer showed a new protein structure with a tandem arrangement consisting of at least three domains and presumably one more. The rod domain had a long hairpin of twisted coiled-coil that extended to one end. The second domain adjacent to the rod alpha-helical domain was globular and constructed by a cluster of eight short beta-sheets. The third domain located distal to the alpha helical rod was globular and composed of seven short beta-sheets and one short alpha-helix. The 13-mer was shaped like a woven rattan cylinder with a large internal tubular space and widely opened flared ends. The 6-mer and 7-mer had a funnel-like structure consisting of a tubular rod at one side and a widely opened flared funnel top at the other side. Based on these results, we constructed a model of the MexAB-OprM pump assembly. The three pairs of MexA dimers interacted with the periplasmic alpha-barrel domain of OprM via the alpha-helical hairpin, the second domain interacted with both MexB and OprM at their contact site, and the third and disordered domains probably interacted with the distal domain of MexB. In this fashion, the MexA subunit connected MexB and OprM, indicating that MexA is the membrane bridge protein. PMID- 15117958 TI - 90-kDa ribosomal S6 kinase is a direct target for the nuclear fibroblast growth factor receptor 1 (FGFR1): role in FGFR1 signaling. AB - Fibroblast growth factor receptor 1 (FGFR1) is a transmembrane protein capable of transducing stimulation by secreted FGFs. In addition, newly synthesized FGFR1 enters the nucleus in response to cellular stimulation and during development. Nuclear FGFR1 can transactivate CRE (cAMP responsive element), activate CRE binding protein (CREB)-binding protein (CBP) and gene activities causing cellular growth and differentiation. Here, a yeast two-hybrid assay was performed to identify FGFR1-binding proteins and the mechanism of nuclear FGFR1 action. Ten FGFR1-binding proteins were identified. Among the proteins detected with the intracellular FGFR1 domain was a 90-kDa ribosomal S6 kinase (RSK1), a regulator of CREB, CBP, and histone phosphorylation. FGFR1 bound to the N-terminal region of RSK1. The FGFR1-RSK1 interaction was confirmed by co-immunoprecipitation and colocalization in the nucleus and cytoplasm of mammalian cells. Predominantly nuclear FGFR1-RSK1 interaction was observed in the rat brain during neurogenesis and in cAMP-stimulated cultured neural cells. In TE671 cells, transfected FGFR1 colocalized and coimmunoprecipitated, almost exclusively, with nuclear RSK1. Nuclear RSK1 kinase activity and RSK1 activation of CREB were enhanced by transfected FGFR1. In contrast, kinase-deleted FGFR1 (TK-), which did not bind to RSK1 failed to stimulate nuclear RSK1 activity or RSK1 activation of CREB. Kinase inactive FGFR1 (K514A) bound effectively to nuclear RSK1, but it failed to stimulate RSK1. Thus, active FGFR1 kinase regulates the functions of nuclear RSK1. The interaction of nuclear FGFR1 with pluripotent RSK1 offers a new mechanism through which FGFR1 may control fundamental cellular processes. PMID- 15117959 TI - Herpes simplex virus type 1 capsid protein VP26 interacts with dynein light chains RP3 and Tctex1 and plays a role in retrograde cellular transport. AB - Cytoplasmic dynein is the major molecular motor involved in minus-end-directed cellular transport along microtubules. There is increasing evidence that the retrograde transport of herpes simplex virus type 1 along sensory axons is mediated by cytoplasmic dynein, but the viral and cellular proteins involved are not known. Here we report that the herpes simplex virus outer capsid protein VP26 interacts with dynein light chains RP3 and Tctex1 and is sufficient to mediate retrograde transport of viral capsids in a cellular model. A library of herpes simplex virus capsid and tegument structural genes was constructed and tested for interactions with dynein subunits in a yeast two-hybrid system. A strong interaction was detected between VP26 and the homologous 14-kDa dynein light chains RP3 and Tctex1. In vitro pull-down assays confirmed binding of VP26 to RP3, Tctex1, and intact cytoplasmic dynein complexes. Recombinant herpes simplex virus capsids were constructed either with or without VP26. In pull-down assays VP26+ capsids bound to RP3; VP26-capsids did not. To investigate intracellular transport, the recombinant viral capsids were microinjected into living cells and incubated at 37 degrees C. After 1 h VP26+ capsids were observed to co-localize with RP3, Tctex1, and microtubules. After 2 or 4 h VP26+ capsids had moved closer to the cell nucleus, whereas VP26-capsids remained in a random distribution. We propose that VP26 mediates binding of incoming herpes simplex virus capsids to cytoplasmic dynein during cellular infection, through interactions with dynein light chains. PMID- 15117960 TI - Nerve growth factor promotes the survival of sympathetic neurons through the cooperative function of the protein kinase C and phosphatidylinositol 3-kinase pathways. AB - The signaling pathways activated by nerve growth factor (NGF) that account for its ability to promote the survival of neurons are not completely understood. Phosphatidylinositol 3-kinase (PI3K) is critical for the survival of several cell types, including neurons. To determine whether additional signaling pathways cooperate with PI3K to promote survival, we examined other pathways known to be activated by NGF. NGF activated protein kinases C (PKCs) in sympathetic neurons, and pharmacologic PKC activation rescued neurons from apoptosis induced by the withdrawal of NGF. Inhibition of PKCs did not inhibit the survival of NGF maintained neurons. Similarly, inhibition of PI3K caused only a modest attrition of neurons in the presence of NGF. In contrast, the simultaneous inhibition of both PKCs and PI3K induced the apoptotic death of NGF-maintained sympathetic neurons. Inhibition of both PI3K and PKCs promoted the expression and phosphorylation of the proapoptotic transcription factor c-Jun, indicating that these pathways inhibit programmed cell death at the stage of proapoptotic gene expression. In culture conditions under which PI3K inhibition alone kills NGF maintained neurons, PKC inhibition also led to a significant loss of viability, indicating that both pathways are required. Therefore, PKC and PI3K, regardless of the culture conditions, cooperate to promote the NGF-dependent survival of sympathetic neurons. PMID- 15117961 TI - Nestin is a potential mediator of malignancy in human neuroblastoma cells. AB - Amplification of the N-myc proto-oncogene signifies aggressive behavior in human neuroblastoma. Likewise, overexpression of the intermediate filament nestin, a neuroectodermal stem cell marker, is linked to increased aggressiveness in several nervous system tumors. We investigated the interaction of these two proteins in human neuroblastoma cells. Neuroblastic cell variants with high levels of N-Myc protein have significantly higher nestin protein levels than non amplified cell lines, suggesting that the transcription factor N-Myc may regulate nestin expression. Stable transfection of a nestin antisense sequence into neuroblastic, N-myc-amplified, LA1-55n cells results in a 2-fold reduction in nestin protein without altering N-Myc expression. However, cell functions attributed to N-Myc (growth rate, anchorage-independent growth, and motility) all decrease significantly. Transfection studies that modulate N-Myc levels also result in commensurate changes in nestin mRNA and protein amounts as well as in cell proliferation and motility. Thus, nestin appears to be downstream of and regulated by N-Myc. Gel mobility shift assays show that N-Myc binds specifically to E-box sequences in the regulatory second intron of the nestin gene and nuclear run-off studies show that increases in N-Myc protein up-regulate nestin transcription rate. Subcellular fractionation and immunoblot studies indicate that nestin is present in the nucleus as well as in the cytoplasm of neuroblastoma cell lines. Finally, DNA cross-linking experiments show that nestin binds DNA in N-myc-amplified N-type cell lines. Thus, nestin may be one mediator of N-myc-associated tumor aggressiveness of human neuroblastoma. PMID- 15117963 TI - Crystal structures of Staphylococcus aureus sortase A and its substrate complex. AB - The cell wall envelope of staphylococci and other Gram-positive pathogens is coated with surface proteins that interact with human host tissues. Surface proteins of Staphylococcus aureus are covalently linked to the cell wall envelope by a mechanism requiring C-terminal sorting signals with an LPXTG motif. Sortase (SrtA) cleaves surface proteins between the threonine (T) and the glycine (G) of the LPXTG motif and catalyzes the formation of an amide bond between threonine at the C-terminal end of polypeptides and cell wall cross-bridges. The active site architecture and catalytic mechanism of sortase A has hitherto not been revealed. Here we present the crystal structures of native SrtA, of an active site mutant of SrtA, and of the mutant SrtA complexed with its substrate LPETG peptide and describe the substrate binding pocket of the enzyme. Highly conserved proline (P) and threonine (T) residues of the LPXTG motif are held in position by hydrophobic contacts, whereas the glutamic acid residue (E) at the X position points out into the solvent. The scissile T-G peptide bond is positioned between the active site Cys(184) and Arg(197) residues and at a greater distance from the imidazolium side chain of His(120). All three residues, His(120), Cys(184), and Arg(197), are conserved in sortase enzymes from Gram-positive bacteria. Comparison of the active sites of S. aureus sortase A and sortase B provides insight into substrate specificity and suggests a universal sortase-catalyzed mechanism of bacterial surface protein anchoring in Gram-positive bacteria. PMID- 15117962 TI - The LIM-only proteins FHL2 and FHL3 interact with alpha- and beta-subunits of the muscle alpha7beta1 integrin receptor. AB - FHL1, FHL2, and FHL3 are members of the four and one-half LIM domain protein subclass that are expressed in striated muscles. Here we show that FHL2 and FHL3 are novel alpha(7)beta(1) integrin-interacting proteins. They bind both the alpha and the beta-subunit as well as different splice isoforms. The minimal binding sites for FHL2 and FHL3 on beta(1A)-chain overlap, whereas on alpha(7A) and alpha(7B) subunits they are situated adjacent. Determining the binding sites for integrins on FHL2 or FHL3 revealed that the suprastructure of the whole molecule is important for these associations, rather than any single LIM domain. Immunofluorescence studies with cells expressing full-length FHL proteins or their deletion mutants showed that FHL2 and FHL3 but not FHL1 colocalize with integrins at cell adhesion sites. Further, their recruitment to the membrane results from binding to either the alpha- or the beta-chain of the integrin receptor. The association of FHL2 or FHL3 with integrin receptors neither influences attachment of cells to different substrates nor changes their migration capacity. However, in cardiac and skeletal muscles, FHL2 and FHL3, respectively, are colocalized with alpha(7)beta(1) integrin receptor at the periphery of Z-discs, suggesting a role in mechanical stabilization of muscle cells. PMID- 15117965 TI - pH dependence of tryptophan synthase catalytic mechanism: I. The first stage, the beta-elimination reaction. AB - The pyridoxal 5'-phosphate-dependent beta-subunit of the tryptophan synthase alpha(2)beta(2) complex catalyzes the condensation of L-serine with indole to form L-tryptophan. The first stage of the reaction is a beta-elimination that involves a very fast interconversion of the internal aldimine in a highly fluorescent L-serine external aldimine that decays, via the alpha-carbon proton removal and beta-hydroxyl group release, to the alpha-aminoacrylate Schiff base. This reaction is influenced by protons, monovalent cations, and alpha-subunit ligands that modulate the distribution between open and closed conformations. In order to identify the ionizable residues that might assist catalysis, we have investigated the pH dependence of the rate of the external aldimine decay by rapid scanning UV-visible absorption and single wavelength fluorescence stopped flow. In the pH range 6-9, the reaction was found to be biphasic with the first phase (rate constants k(1)) accounting for more than 70% of the signal change. In the absence of monovalent cations or in the presence of sodium and potassium ions, the pH dependence of k(1) exhibits a bell shaped profile characterized by a pK(a1) of about 6 and a pK(a2) of about 9, whereas in the presence of cesium ions, the pH dependence exhibits a saturation profile characterized by a single pK(a) of 9. The presence of the allosteric effector indole acetylglycine increases the rate of reaction without altering the pH profile and pK(a) values. By combining structural information for the internal aldimine, the external aldimine, and the alpha-aminoacrylate with kinetic data on the wild type enzyme and beta-active site mutants, we have tentatively assigned pK(a1) to betaAsp-305 and pK(a2) to betaLys-87. The loss of pK(a1) in the presence of cesium ions might be due to a shift to lower values, caused by the selective stabilization of a closed form of the beta-subunit. PMID- 15117964 TI - Gastrointestinally distributed UDP-glucuronosyltransferase 1A10, which metabolizes estrogens and nonsteroidal anti-inflammatory drugs, depends upon phosphorylation. AB - Among gastrointestinal distributed isozymes encoded at the UGT1 locus, UDP glucuronosyltransferase 1A10 (UGT1A10) metabolizes a number of important chemicals. Similar to broad conversion of phytoestrogens (Basu, N. K., Ciotti, M., Hwang, M. S., Kole, L., Mitra, P. S., Cho, J. W., and Owens, I. S. (2004) J. Biol. Chem. 279, 1429-1441), UGT1A10 metabolized estrogens and their derivatives, whereas UGT1A1, -1A3, -1A7, and -1A8 differentially exhibited reduced activity toward the same. UGT1A10 compared with UGT1A7, -1A8, and -1A3 generally exhibited high activity toward acidic nonsteroidal anti-inflammatory drugs and natural benzaldehyde derivatives, while UGT1A3 metabolized most efficiently aromatic transcinnamic acids known to be generated from flavonoid glycosides by microflora in the lower gastrointestinal tract. Finally UGT1A10, -1A7, -1A8, and -1A3 converted plant-based salicylic acids; methylsalicylic acid was transformed at high levels, and acetylsalicylic (aspirin) and salicylic acid were transformed at moderate to low levels. Atypically UGT1A10 transformed estrogens between pH 6 and 8 but acidic structures preferentially at pH 6.4. Furthermore evidence indicates UGT1A10 expressed in COS-1 cells depends upon phosphorylation; UGT1A10 versus its single, double, and triple mutants at three predicted protein kinase C phosphorylation sites incorporated [(33)P]-orthophosphate and showed a progressive decrease with no detectable label or activity for the triple T73A/T202A/S432G-1A10 mutant. Single and double mutants revealed either null/full activity or null/additive activity, respectively. Additionally UGT1A10-expressing cultures glucuronidated 17beta-[(14)C]estradiol, whereas cultures containing null mutants at protein kinase C sites showed no estrogen conversion. Importantly UGT1A10 in cells supported 10-fold higher glucuronidation of 17beta-estradiol than UGT1A1. In summary, our results suggest gastrointestinally distributed UGT1A10 is important for detoxifying estrogens/phytoestrogens and aromatic acids with complementary activity by UGT1A7, -1A8, -1A3, and/or -1A1 evidently dependent upon phosphorylation. PMID- 15117966 TI - Poly(A)+ RNAs roam the cell nucleus and pass through speckle domains in transcriptionally active and inactive cells. AB - Many of the protein factors that play a role in nuclear export of mRNAs have been identified, but still little is known about how mRNAs are transported through the cell nucleus and which nuclear compartments are involved in mRNA transport. Using fluorescent 2'O-methyl oligoribonucleotide probes, we investigated the mobility of poly(A)+ RNA in the nucleoplasm and in nuclear speckles of U2OS cells. Quantitative analysis of diffusion using photobleaching techniques revealed that the majority of poly(A)+ RNA move throughout the nucleus, including in and out of speckles (also called SC-35 domains), which are enriched for splicing factors. Interestingly, in the presence of the transcription inhibitor 5,6-dichloro-1-beta D-ribofuranosylbenzimidazole, the association of poly(A)+ RNA with speckles remained dynamic. Our results show that RNA movement is energy dependent and that the proportion of nuclear poly(A)+ RNA that resides in speckles is a dynamic population that transiently interacts with speckles independent of the transcriptional status of the cell. Rather than the poly(A)+ RNA within speckles serving a stable structural role, our findings support the suggestion of a more active role of these regions in nuclear RNA metabolism and/or transport. PMID- 15117967 TI - Fascin-mediated propulsion of Listeria monocytogenes independent of frequent nucleation by the Arp2/3 complex. AB - Actin-dependent propulsion of Listeria monocytogenes is thought to require frequent nucleation of actin polymerization by the Arp2/3 complex. We demonstrate that L. monocytogenes motility can be separated into an Arp2/3-dependent nucleation phase and an Arp2/3-independent elongation phase. Elongation-based propulsion requires a unique set of biochemical factors in addition to those required for Arp2/3-dependent motility. We isolated fascin from brain extracts as the only soluble factor required in addition to actin during the elongation phase for this type of movement. The nucleation reaction assembles a comet tail of branched actin filaments directly behind the bacterium. The elongation-based reaction generates a hollow cylinder of parallel bundles that attach along the sides of the bacterium. Bacteria move faster in the elongation reaction than in the presence of Arp2/3, and the rate is limited by the concentration of G-actin. The biochemical and structural differences between the two motility reactions imply that each operates through distinct biochemical and biophysical mechanisms. PMID- 15117968 TI - Sequential exocytosis of insulin granules is associated with redistribution of SNAP25. AB - We have investigated sequential exocytosis in beta cells of intact pancreatic islets with the use of two-photon excitation imaging of a polar fluorescent tracer, sulforhodamine B, and a fusion protein comprising enhanced cyan fluorescent protein (ECFP) and the SNARE protein SNAP25 (synaptosome-associated protein of 25 kD) transfected with an adenoviral vector. Sequential exocytosis was found to account for <10% of exocytic events in beta cells stimulated either with glucose under various conditions or by photolysis of a caged-Ca2+ compound. Multigranular exocytosis, in which granule-to-granule fusion occurs before exocytosis, was rarely found. We detected redistribution of ECFP-SNAP25 from the plasma membrane into the membrane of the fused granule occurred in a large proportion (54%) of sequential exocytic events but in only a small fraction (5%) of solitary fusion events. Removal of cholesterol in the plasma membrane by methyl-beta-cyclodextrin facilitated both redistribution of ECFP-SNAP25 and sequential exocytosis by threefold. These observations support the hypothesis that SNAP25 is a plasma membrane factor that is responsible for sequential exocytosis. PMID- 15117969 TI - Autocrine CSF-1R activation promotes Src-dependent disruption of mammary epithelial architecture. AB - Elevated coexpression of colony-stimulating factor receptor (CSF-1R) and its ligand, CSF-1, correlates with invasiveness and poor prognosis of a variety of epithelial tumors (Kacinski, B.M. 1995. Ann. Med. 27:79-85). Apart from recruitment of macrophages to the tumor site, the mechanisms by which CSF-1 may potentiate invasion are poorly understood. We show that autocrine CSF-1R activation induces hyperproliferation and a profound, progressive disruption of junctional integrity in acinar structures formed by human mammary epithelial cells in three-dimensional culture. Acini coexpressing receptor and ligand exhibit a dramatic relocalization of E-cadherin from the plasma membrane to punctate intracellular vesicles, accompanied by its loss from the Triton insoluble fraction. Interfering with Src kinase activity, either by pharmacological inhibition or mutation of the Y561 docking site on CSF-1R, prevents E-cadherin translocation, suggesting that CSF-1R disrupts cell adhesion by uncoupling adherens junction complexes from the cytoskeleton and promoting cadherin internalization through a Src-dependent mechanism. These findings provide a mechanistic basis whereby CSF-1R could contribute to invasive progression in epithelial cancers. PMID- 15117970 TI - Mesenchymal-epithelial interactions in the skin: increased expression of dickkopf1 by palmoplantar fibroblasts inhibits melanocyte growth and differentiation. AB - We investigated whether or not the topographic regulation of melanocyte differentiation is determined by mesenchymal-epithelial interactions via fibroblast-derived factors. The melanocyte density in palmoplantar human skin (i.e., skin on the palms and the soles) is five times lower than that found in nonpalmoplantar sites. Palmoplantar fibroblasts significantly suppressed the growth and pigmentation of melanocytes compared with nonpalmoplantar fibroblasts. Using cDNA microarray analysis, fibroblasts derived from palmoplantar skin expressed high levels of dickkopf 1 (DKK1; an inhibitor of the canonical Wnt signaling pathway), whereas nonpalmoplantar fibroblasts expressed higher levels of DKK3. Transfection studies revealed that DKK1 decreased melanocyte function, probably through beta-catenin-mediated regulation of microphthalmia-associated transcription factor activity, which in turn modulates the growth and differentiation of melanocytes. Thus, our results provide a basis to explain why skin on the palms and the soles is generally hypopigmented compared with other areas of the body, and might explain why melanocytes stop migrating in the palmoplantar area during human embryogenesis. PMID- 15117971 TI - Somatic hypermutation is limited by CRM1-dependent nuclear export of activation induced deaminase. AB - Somatic hypermutation (SHM) and class switch recombination (CSR) are initiated in activated B lymphocytes by activation-induced deaminase (AID). AID is thought to make lesions in DNA by deaminating cytidine residues in single-stranded DNA exposed by RNA polymerase during transcription. Although this must occur in the nucleus, AID is found primarily in the cytoplasm. Here we show that AID is actively excluded from the nucleus by an exportin CRM1-dependent pathway. The AID nuclear export signal (NES) is found at the carboxyl terminus of AID in a region that overlaps a sequence required for CSR but not SHM. We find that AID lacking a functional NES causes more hypermutation of a nonphysiologic target gene in transfected fibroblasts. However, the NES does not impact on the rate of mutation of immunoglobulin genes in B lymphocytes, suggesting that the AID NES does not limit AID activity in these cells. PMID- 15117972 TI - Defective suppressor function of human CD4+ CD25+ regulatory T cells in autoimmune polyglandular syndrome type II. AB - In autoimmune polyglandular syndromes (APS), several organ-specific autoimmune diseases are clustered. Although APS type I is caused by loss of central tolerance, the etiology of APS type II (APS-II) is currently unknown. However, in several murine models, depletion of CD4(+) CD25(+) regulatory T cells (T(regs)) causes a syndrome resembling human APS-II with multiple endocrinopathies. Therefore, we hypothesized that loss of active suppression in the periphery could be a hallmark of this syndrome. T(regs) from peripheral blood of APS-II, control patients with single autoimmune endocrinopathies, and normal healthy donors showed no differences in quantity (except for patients with isolated autoimmune diseases), in functionally important surface markers, or in apoptosis induced by growth factor withdrawal. Strikingly, APS-II T(regs) were defective in their suppressive capacity. The defect was persistent and not due to responder cell resistance. These data provide novel insights into the pathogenesis of APS-II and possibly human autoimmunity in general. PMID- 15117973 TI - A critical temporal window for selectin-dependent CD4+ lymphocyte homing and initiation of late-phase inflammation in contact sensitivity. AB - Contact sensitivity (CS) is an inflammatory disorder characterized by early and late phases of leukocyte recruitment. We used a noninvasive intravital microscopy technique allowing for the direct visualization of leukocyte rolling and adhesion on blood vessel endothelium. By blocking specific adhesion molecules, we elucidated the molecular mechanisms mediating early leukocyte recruitment to be E and P-selectin and demonstrated that leukocyte recruitment in the late phase had a different adhesive profile (mainly alpha(4)-integrin). Complete blockade of E- and P-selectin within the first 2 h of leukocyte-endothelial cell interactions (but not later) eliminated selectin-independent leukocyte recruitment at 24 h. Despite the predominance of neutrophils in the early phase, specific elimination of CD4(+) lymphocytes in the early phase eliminated the late response. CD4(+) lymphocytes homed to skin via E- and P-selectin within the early phase and induced the late phase response. Addition of these same CD4(+) lymphocytes 2 h after antigen challenge was too late for these cells to home to the skin and induce late phase responses. Our data clearly demonstrate that the antigen challenged microenvironment is only accessible to CD4(+) lymphocytes for the first 2 h, and that this process is essential for the subsequent recruitment of other leukocyte populations in late phase responses. PMID- 15117974 TI - Arylamine N-acetyltransferase is required for synthesis of mycolic acids and complex lipids in Mycobacterium bovis BCG and represents a novel drug target. AB - Mycolic acids represent a major component of the unique cell wall of mycobacteria. Mycolic acid biosynthesis is inhibited by isoniazid, a key frontline antitubercular drug that is inactivated by mycobacterial and human arylamine N-acetyltransferase (NAT). We show that an in-frame deletion of Mycobacterium bovis BCG nat results in delayed entry into log phase, altered morphology, altered cell wall lipid composition, and increased intracellular killing by macrophages. In particular, deletion of nat perturbs biosynthesis of mycolic acids and their derivatives and increases susceptibility of M. bovis BCG to antibiotics that permeate the cell wall. Phenotypic traits are fully complemented by introduction of Mycobacterium tuberculosis nat. We infer from our findings that NAT is critical to normal mycolic acid synthesis and hence other derivative cell wall components and represents a novel target for antituberculosis therapy. In addition, this is the first report of an endogenous role for NAT in mycobacteria. PMID- 15117975 TI - Legionella subvert the functions of Rab1 and Sec22b to create a replicative organelle. AB - Legionella pneumophila is a bacterial pathogen that infects eukaryotic host cells and replicates inside a specialized organelle that is morphologically similar to the endoplasmic reticulum (ER). To better understand the molecular mechanisms governing transport of the Legionella-containing vacuole (LCV), we have identified host proteins that participate in the conversion of the LCV into a replicative organelle. Our data show that Rab1 is recruited to the LCV within minutes of uptake. Rab1 recruitment to the LCV precedes remodeling of this compartment by ER-derived vesicles. Genetic inhibition studies demonstrate that Rab1 is important for the recruitment of ER-derived vesicles to the LCV and that inhibiting Rab1 function abrogates intracellular growth of Legionella. Morphological studies indicate that the Sec22b protein is located on ER-derived vesicles recruited to the LCV and that Sec22b is delivered to the LCV membrane. Sec22b function was found to be important for biogenesis of the specialized organelle that supports Legionella replication. These studies demonstrate that Legionella has the ability to subvert Rab1 and Sec22b function to facilitate the transport and fusion of ER-derived vesicles with the LCV, resulting in the formation of a specialized organelle that can support bacterial replication. PMID- 15117976 TI - CD43 regulation of T cell activation is not through steric inhibition of T cell APC interactions but through an intracellular mechanism. AB - CD43 is a large heavily glycosylated protein highly expressed on T cells and actively excluded from the immunological synapse through interactions with ezrin radixin-moesin proteins. Due to its size and charge, it has been proposed that the CD43 ectodomain acts as a physical barrier to T cell-APC interactions. We have addressed this hypothesis by studying the effect of reconstituting CD43 mutants into the hyperproliferative CD43(-/-) T cells. Reintroduction of full length CD43 reversed the CD43(-/-) T cell hyperproliferation. Interestingly, despite the lack of exclusion from the interaction site, a mutant containing the CD43 ectodomain on a glycosylphosphatidylinositol linkage was ineffective. Additionally, T cell-APC conjugate formation was not affected by this ectodomain only construct. In contrast, CD43(-/-) T cell hyperproliferation was reversed by an intracellular-only CD43 fused to the small ectodomain of hCD16. Mutation of this intracellular-only CD43 such that it could not move from the T cell-APC contact site had no further affect on proliferation than the moveable CD43 but did dramatically reduce interleukin-2 production. Thus, the exclusion of the CD43 intracellular region from the immunological synapse is required for CD43 regulation of interleukin-2 production, but the presence of the cytoplasmic tail, independent of its location, is sufficient to reverse CD43(-/-) T cell hyperproliferation. PMID- 15117977 TI - The ethics of early stopping rules: who is protecting whom? PMID- 15117978 TI - Phase I and pharmacokinetic study of the ribonucleotide reductase inhibitor, 3 aminopyridine-2-carboxaldehyde thiosemicarbazone, administered by 96-hour intravenous continuous infusion. AB - PURPOSE: 3-aminopyridine-2-carboxaldehyde thiosemicarbazone (3-AP; Triapine; Vion Pharmaceuticals Inc, New Haven, CT) is a potent inhibitor of ribonucleotide reductase, with activity in preclinical tumor model systems. A phase I trial was initiated to determine the dose-limiting toxicities, maximum-tolerated dose, and pharmacokinetics of a 96-hour intravenous (IV) continuous infusion in patients with advanced cancer. PATIENTS AND METHODS: Initially, courses were administered every 3 weeks, using an accelerated titration design. Subsequently, courses were administered every 2 weeks, and the dose was escalated in cohorts of three to six patients. RESULTS: Twenty-one patients were enrolled, seven on the every-3-week schedule and 14 on the every-other-week schedule. Three of six patients at 160 mg/m(2)/d developed dose-limiting toxicities including neutropenia, hyperbilirubinemia, and nausea or vomiting. Based on these initial results, the dose for 3-AP was re-escalated beginning at 80 mg/m(2)/d but administered every 2 weeks. At 120 mg/m(2)/d, three of seven patients had dose-limiting but reversible asthenia, hyperbilirubinemia, and azotemia or acidosis; however, in the case of renal and hepatic adverse events, the events were related to pre-existing borderline abnormal organ function. Therefore, the recommended phase II dose for 3-AP administered by 96-hour IV infusion is 120 mg/m(2)/d every 2 weeks. Detailed pharmacokinetic studies demonstrated linear kinetics up to 160 mg/m(2), with substantial inter-patient variability. There was no correlation between dose and clearance (R(2) = 0.0137). There were no objective responses, but there was prolonged stabilization of disease or decreases in serum tumor markers associated with stable disease in four patients. CONCLUSION: The 96-hour infusion of 3-AP is safe and well tolerated at the recommended phase II doses. Phase II trials of Triapine are ongoing. PMID- 15117979 TI - Prognostic value of proliferation, apoptosis, defective DNA mismatch repair, and p53 overexpression in patients with resected Dukes' B2 or C colon cancer: a North Central Cancer Treatment Group Study. AB - PURPOSE: Molecular studies of colon cancer have provided insights into pathogenesis, yet it is unclear how important these markers are in predicting prognosis. This study investigated the prognostic significance of TUNEL, bcl-2, p53, proliferation marker Ki-67 and DNA mismatch repair (MMR) status in patients with Dukes' stage B2 and C colorectal adenocarcinomas. PATIENTS AND METHODS: Tumor tissue from 366 patients (75% Dukes' C, 25% Dukes' B2) from four randomized North Central Cancer Treatment Group phase III surgical adjuvant trials were used. Eighty-one percent of patients received adjuvant treatment, which was primarily fluorouracil (FU) based (90%). Tumor location was predominantly (87%) the colon. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), Ki-67, p53, bcl-2, and MMR were assayed using immunohistochemistry. Stage, grade, MMR, Ki-67, and previously determined flow cytometry markers (ploidy and S phase) were explored for associations with each other and with overall survival (OS) and disease-free survival (DFS). RESULTS: Univariately, stage B2, low grade, diploid, Ki-67 more than 27%, normal p53, and FU-based adjuvant treatment were significantly associated with improved OS and DFS (P <.05). After adjusting for stage, grade, and ploidy in multivariate analysis, Ki 67 remained significantly related to both OS and DFS (P <.01). Active FU-based adjuvant treatment was significant only for OS in this multivariate model. Neither bcl-2 nor TUNEL were significant. CONCLUSION: This retrospective study indicates that Ki-67 and ploidy may have stronger prognostic impact on OS and DFS than other parameters investigated after adjusting for stage and tumor grade. Prospective studies to elucidate the mechanism and prognostic significance of these findings are necessary. PMID- 15117980 TI - Randomized phase III trial of pemetrexed versus docetaxel in patients with non small-cell lung cancer previously treated with chemotherapy. AB - PURPOSE: To compare the efficacy and toxicity of pemetrexed versus docetaxel in patients with advanced non-small-cell lung cancer (NSCLC) previously treated with chemotherapy. PATIENTS AND METHODS: Eligible patients had a performance status 0 to 2, previous treatment with one prior chemotherapy regimen for advanced NSCLC, and adequate organ function. Patients received pemetrexed 500 mg/m(2) intravenously (i.v.) day 1 with vitamin B(12), folic acid, and dexamethasone or docetaxel 75 mg/m(2) i.v. day 1 with dexamethasone every 21 days. The primary end point was overall survival. RESULTS: Five hundred seventy-one patients were randomly assigned. Overall response rates were 9.1% and 8.8% (analysis of variance P =.105) for pemetrexed and docetaxel, respectively. Median progression free survival was 2.9 months for each arm, and median survival time was 8.3 versus 7.9 months (P = not significant) for pemetrexed and docetaxel, respectively. The 1-year survival rate for each arm was 29.7%. Patients receiving docetaxel were more likely to have grade 3 or 4 neutropenia (40.2% v 5.3%; P <.001), febrile neutropenia (12.7% v 1.9%; P <.001), neutropenia with infections (3.3% v 0.0%; P =.004), hospitalizations for neutropenic fever (13.4% v 1.5%; P <.001), hospitalizations due to other drug related adverse events (10.5% v 6.4%; P =.092), use of granulocyte colony-stimulating factor support (19.2% v 2.6%, P <.001) and all grade alopecia (37.7% v 6.4%; P <.001) compared with patients receiving pemetrexed. CONCLUSION: Treatment with pemetrexed resulted in clinically equivalent efficacy outcomes, but with significantly fewer side effects compared with docetaxel in the second-line treatment of patients with advanced NSCLC and should be considered a standard treatment option for second line NSCLC when available. PMID- 15117981 TI - First-line chemotherapy with cisplatin plus fractionated temozolomide in recurrent glioblastoma multiforme: a phase II study of the Gruppo Italiano Cooperativo di Neuro-Oncologia. AB - PURPOSE: Cisplatin and temozolomide (TMZ) are active in glioblastoma multiforme (GBM), with different profiles of toxicity. A bid regimen of TMZ achieves a strong inhibition of O(6)-alkylguanine DNA-alkyl transferase (AGAT), and cisplatin reduces AGAT activity in vitro, suggesting a possible synergic interaction. The primary end point of the present multicenter phase II study was progression-free survival (PFS) at 6 months (PFS-6); secondary end points included response, toxicity, and overall survival. PATIENTS AND METHODS: Chemotherapy-naive patients with GBM who experienced disease recurrence or progression after surgery and standard radiotherapy were eligible. Chemotherapy cycles consisted of cisplatin 75 mg/m(2) on day 1, TMZ 130 mg/m(2) bolus followed by nine doses of 70 mg/m(2) every 12 hours (total of 5 days) from day 2 every 4 weeks. In the absence of hematologic toxicity, TMZ was escalated to 1,000 mg/m(2) in 5 days. RESULTS: A total of 50 patients (median age, 53.4 years; range, 27 to 70 years; median Karnofsky performance status, 80; range, 60 to 100) were accrued in the study. PFS-6 was 34% (95% CI, 23% to 50%), and PFS-12 was 4% (95% CI, 0.3% to 16%). Median PFS was 18.4 weeks (95% CI, 13 to 25.9 weeks). Among 49 assessable patients, one complete response and nine partial responses were obtained, with an overall response rate of 20.4% (95% CI, 7.7% to 33%). Among 203 treatment cycles delivered, the most common grade 3 or grade 4 events included granulocytopenia in 7.9% of cycles, thrombocytopenia in 4%, and neurologic toxicity in three patients (6%). CONCLUSION: The new cisplatin plus bid TMZ regimen appears active in chemotherapy-naive patients with recurrent GBM and incurs an acceptable toxicity. PMID- 15117982 TI - Comparison of fulvestrant versus tamoxifen for the treatment of advanced breast cancer in postmenopausal women previously untreated with endocrine therapy: a multinational, double-blind, randomized trial. AB - PURPOSE: To evaluate the efficacy and tolerability of fulvestrant (Faslodex; AstraZeneca Pharmaceuticals LP, Wilmington, DE), a new estrogen receptor (ER) antagonist that downregulates ER and has no agonist effects, versus tamoxifen, an antiestrogen with agonist and antagonist effects, for the treatment of advanced breast cancer in postmenopausal women. PATIENTS AND METHODS: In this multicenter, double-blind, randomized trial, patients with metastatic/locally advanced breast cancer previously untreated for advanced disease were randomly assigned to receive either fulvestrant (250 mg, via intramuscular injection, once monthly; n = 313) or tamoxifen (20 mg, orally, once daily; n = 274). Patients' tumors were positive for ER (ER+) and/or progesterone receptor (PgR+), or had an unknown receptor status. RESULTS: At a median follow-up of 14.5 months, there was no significant difference between fulvestrant and tamoxifen for the primary end point of time to progression (TTP; median TTP, 6.8 months and 8.3 months, respectively; hazard ratio, 1.18; 95% CI, 0.98 to 1.44; P =.088). In a prospectively planned subset analysis of patients with known ER+ and/or PgR+ tumors ( approximately 78%), median TTP was 8.2 months for fulvestrant and 8.3 months for tamoxifen (hazard ratio, 1.10; 95% CI, 0.89 to 1.36; P =.39). The objective response rate for the overall population was 31.6% with fulvestrant and 33.9% with tamoxifen, and 33.2% and 31.1%, respectively, in the known hormone receptor-positive subgroup. Both treatments were well tolerated. CONCLUSION: In the overall population, between-group differences in efficacy end points favored tamoxifen, and statistical noninferiority of fulvestrant could not be demonstrated. However, in patients with hormone receptor-positive tumors, fulvestrant had similar efficacy to tamoxifen and was well tolerated. PMID- 15117983 TI - Clinical relevance of different sequencing of doxorubicin and cyclophosphamide, methotrexate, and Fluorouracil in operable breast cancer. AB - PURPOSE: To assess the clinical relevance of different sequences of doxorubicin (DOX) and cyclophosphamide, methotrexate, and fluorouracil (CMF) in patients with operable breast cancer at risk of disease relapse. PATIENTS AND METHODS: Two randomized trials were activated in the early 1980s. The first study, in patients with one to three involved nodes, was intended to assess the effectiveness of intravenous (i.v.) CMF given every 3 weeks for 12 courses versus eight courses of the same CMF regimen followed by four courses of full-dose DOX (CMF-->DOX). The second study, in patients with more than three involved nodes, compared four courses of full-dose DOX sequentially followed by eight courses of i.v. CMF (DOX- >CMF) versus alternating two courses of the same CMF regimen with one course of DOX (CMF/DOX) for a total of 12 courses. RESULTS: After a median observation of 210 months, no statistically significant difference was documented in the first study (relapse-free survival hazard rate [HR], 1.06; total survival HR, 1.03). In contrast, the delivery of DOX first, followed by CMF significantly reduced the risk of disease relapse (HR, 0.68; 95% CI, 0.54 to 0.87; P =.0017) and death (HR, 0.74; 95% CI, 0.57 to 0.95; P =.018) compared with the alternating regimen. CONCLUSION: Anthracycline-containing regimens can further reduce the odds of relapse and death compared with CMF. However, the findings observed in our trials emphasize that the relative merits of anthracycline adjuvant programs also can depend on the modality of administration and must be assessed in properly designed trials in which the magnitude of the benefits can be weighed against potential risks. PMID- 15117984 TI - Phase II trial of trastuzumab followed by weekly paclitaxel/carboplatin as first line treatment for patients with metastatic breast cancer. AB - PURPOSE: To determine the response rate of trastuzumab as first-line therapy in patients with HER-2 overexpressing metastatic breast cancer. To assess the feasibility and toxicity of weekly paclitaxel/carboplatin with or without trastuzumab following initial treatment with trastuzumab. PATIENTS AND METHODS: Sixty-one patients received trastuzumab (8 mg/kg followed by 4 mg/kg/wk) for 8 weeks. Responding patients received 8 additional weeks of trastuzumab (4 mg/kg/wk), and then proceeded to receive trastuzumab (2 mg/kg) in combination with paclitaxel 70 mg/m(2) and carboplatin (area under the curve, 2) weekly for 6 weeks followed by 2 weeks rest. Stable patients after the initial 8 weeks of trastuzumab proceeded to treatment with trastuzumab, paclitaxel, and carboplatin. Patients with disease progression during the initial 8 weeks had the trastuzumab discontinued and were treated with weekly paclitaxel/carboplatin. RESULTS: Weekly paclitaxel/carboplatin with or without trastuzumab was well tolerated. Fifty-two patients were assessable for response and all 61 patients were assessable for survival. Seventeen (33%) of 52 patients experienced a minor/partial response to single-agent trastuzumab and received 8 additional weeks of single-agent trastuzumab. Fifteen (29%) of 52 patients had stable disease and proceeded to receive paclitaxel/carboplatin/trastuzumab. Thirty-one patients with measurable disease were assessable for response after initial single-agent trastuzumab followed by paclitaxel/carboplatin/trastuzumab. An overall response rate of 84% (eight complete responses/18 partial responses), median time to progression of 14.2 months, and median overall survival of 32.2 months was reported with the triplet combination. In the patients treated with paclitaxel/carboplatin alone after disease progression on initial single-agent trastuzumab, an overall response rate of 69% (one complete response/10 partial responses), median time to progression of 8.3 months, and median overall survival of 22.2 months was reported. Median time to progression for all 61 patients is 10 months and the median overall survival is 26.7 months. CONCLUSION: This trial confirms the activity and tolerability of weekly paclitaxel/carboplatin alone or in combination with trastuzumab in women with HER-2 overexpressing metastatic breast cancer. PMID- 15117985 TI - Ten-year outcomes in a population-based cohort of node-negative, lymphatic, and vascular invasion-negative early breast cancers without adjuvant systemic therapies. AB - PURPOSE: To discuss the absolute benefits from adjuvant systemic therapy knowledge of long-term outcomes and baseline risks of relapse and disease specific survival are required. We assessed the 10-year outcomes in a population based cohort of node-negative (N-) lymphovascular negative (LV-) early breast cancers diagnosed from 1989 to 1991 who did not receive adjuvant systemic therapy. METHODS: One thousand one hundred eighty-seven cases of pT(1-2)N(0) LV- breast cancers with a median follow-up of 10.4 years were reviewed. Kaplan-Meier survival curves for relapse free survival (RFS), breast cancer-specific survival (BCSS) and overall survival (OS) were compared with log-rank tests with cohorts stratified for tumor size and grade. RESULTS: The median age of this series was 62 years. Four hundred thirty tumors were < or = 1 cm in diameter (cohort 1), 507 were 1.1-2 cm (cohort 2), and 250 were 2.1 to 5 cm in diameter (cohort 3). The 10 year outcomes for cohorts 1, 2, and 3, respectively, were significantly different: RFS, 82%, 75%, and 66%; BCSS, 92%, 90%, and 77%; and OS, 79%, 78%, and 66%. Tumor grade significantly altered outcome within size cohorts, particularly in pT(1)N(0) breast cancers. CONCLUSION: This study provides detailed information on the continued relapse and breast cancer death rate to 10 years of follow-up. Specifically, without adjuvant systemic therapy, patients with LV-, N - breast cancer had a > or = 25% 10-year risk of relapse and a corresponding 10-year breast cancer death rate of > or = 10% if they had either a grade 3 tumor < or = 1 cm, a grade 2 to 3 tumor from 1.1 to 2 cm, or any grade tumor greater than 2 cm. PMID- 15117986 TI - Incidence of BRCA1 and BRCA2 mutations in young Korean breast cancer patients. AB - PURPOSE: The prevalence of BRCA-associated breast carcinoma in the Korean population has not been evaluated extensively. METHODS: Sixty Korean women who developed breast cancer by age 40 years were studied. Lymphocyte specimens from peripheral blood were processed for BRCA1 and BRCA2 by complete sequencing. Family history through three generations was obtained. Available paraffin embedded tissue blocks were processed for immunohistochemical staining. RESULTS: In the cohort of 60 patients, nine patients with 11 deleterious mutations (six in BRCA1 and five in BRCA2) and seven missense mutations of unknown significance were found. Two patients had deleterious mutations in both BRCA1 and BRCA2 (double mutant). One half of the mutations were novel, and no founder mutations were observed in this cohort. Most of the BRCA-associated patients had no family history of breast and/or ovarian cancer. The expression of HER-2/neu, cyclin D1, and hormone receptors was less common, and p53 overexpression was more common in BRCA-associated tumors. CONCLUSION: The prevalence of BRCA1 and BRCA2 mutations in Korean women with breast cancer at a young age was high. However, the penetrance, as evidenced by the low frequency of breast and ovarian cancers in family members, appears to be low. These data suggest that there may be different genetic and etiologic factors affecting transmission and penetrance of the BRCA genes in Korean patients with breast cancer diagnosed at a young age. PMID- 15117987 TI - Use of the humanized anti-epidermal growth factor receptor monoclonal antibody h R3 in combination with radiotherapy in the treatment of locally advanced head and neck cancer patients. AB - PURPOSE: To evaluate safety and preliminary efficacy of the humanized anti epidermal growth factor receptor monoclonal antibody h-R3 in combination with radiotherapy (RT) in unresectable head and neck cancer patients. Secondary end points were the measurement of h-R3 serum levels and the assessment of the potential mechanisms of antitumor effect on patient biopsies. Anti-idiotypic response to h-R3 was assessed. To predict pharmacologic effect, a mathematical model for antibodies recognizing antigens expressed in tumors and normal tissues was built. PATIENTS AND METHODS: Twenty-four patients with advanced carcinomas of the head and neck received six once-weekly infusions of h-R3 at four dose levels in combination with RT. Pretreatment tumor biopsies were obtained to evaluate epidermal growth factor receptor expression as an enrollment criterion. Second biopsies were taken to evaluate the proliferative activity and angiogenesis in comparison with the pretreatment samples. Patient serum samples were collected to measure h-R3 levels and anti-idiotypic response. RESULTS: The combination of h-R3 and RT was well tolerated. Antibody-related adverse events consisted in infusion reactions. No skin or allergic toxicity appeared. Overall survival significantly increased after the use of the higher antibody doses. Immunohistochemistry studies of tumor specimens before and after treatment revealed that antitumor response correlated with antiproliferative and antiangiogenic effect. One patient developed antibodies to h-R3. The mathematical model predicted that the maximum difference between the area under the curve in tumors and normal tissues is reached when the antibody has intermediate affinity. CONCLUSION: h-R3 is a well tolerated drug that may enhance radiocurability of unresectable head and neck neoplasms. PMID- 15117988 TI - Association of preoperative plasma levels of vascular endothelial growth factor and soluble vascular cell adhesion molecule-1 with lymph node status and biochemical progression after radical prostatectomy. AB - PURPOSE: Angiogenesis is a critical process for cancer progression. We tested whether elevated circulating levels of the angiogenesis-related markers vascular endothelial growth factor (VEGF) and/or soluble vascular cell adhesion molecule-1 (sVCAM-1) are associated with prostate cancer diagnosis, stage, progression, and metastasis. PATIENTS AND METHODS: Plasma levels of VEGF and sVCAM-1 were measured on frozen, archival plasma obtained preoperatively from 215 consecutive patients who underwent radical prostatectomy for clinically localized disease, nine men with untreated prostate cancer metastatic to bones, and 40 healthy men without cancer. RESULTS: Plasma levels of both VEGF and sVCAM-1 were highest in patients with bone metastases (P <.001). VEGF levels were higher in patients with clinically localized disease than in healthy controls (P <.001). VEGF levels were elevated in patients with biopsy and final Gleason sum > or = 7 (P =.036 and P =.020, respectively) and extraprostatic extension (P =.047). Higher preoperative VEGF was independently associated with metastases to lymph nodes (P <.001). Both VEGF and sVCAM-1 were independently associated with biochemical progression after adjustment for the effects of standard preoperative features (P =.014 and P =.039, respectively). VEGF remained independently associated with biochemical progression after adjustment for standard postoperative features (P =.019). CONCLUSION: Plasma levels of VEGF increased incrementally from healthy controls to patients with clinically localized disease to patients with lymph node and skeletal metastases. Higher preoperative VEGF was independently associated with metastases to lymph nodes and biochemical progression after surgery in both pre- and postoperative models. Plasma sVCAM-1 was elevated in men with bone metastases and was associated with biochemical progression in a preoperative model. PMID- 15117989 TI - Influence of biologic markers on the outcome of Hodgkin's lymphoma: a study by the Spanish Hodgkin's Lymphoma Study Group. AB - PURPOSE: Current therapies fail to cure a significant proportion of patients with Hodgkin's lymphoma (HL). Predictive systems for stratification of the disease and selection of treatment based on sets of clinical variables, such as the international prognostic score (IPS), are of relatively small practical value. The predictive use of biologic parameters has so far provided limited and inconsistent results. Here we explore the influence of a set of molecular markers on the outcome of HL. PATIENTS AND METHODS: Forty molecular markers involved in B cell differentiation and activation, signal transduction, cell cycle, and apoptosis control were analyzed in 259 classic HL patient cases by using tissue microarrays. Univariate analysis was performed to evaluate the influence of markers on favorable outcome (complete remission of > 12 months). Significant variables were included in a multivariate logistic regression analysis, and the probability of favorable outcome was estimated. RESULTS: Univariate analysis revealed four molecular markers that predicted outcome, and the multivariate analysis showed p53, Bcl-X(L), and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick-end labeling (TUNEL) to have independent significance. The combination of these factors determined two groups of patients (group I, zero to one factor; group II, two to three factors) with a probability of a favorable outcome of.948 and.687, respectively. A multivariate Cox's model shows that these biologic risk groups have special predictive power in low-IPS patients. CONCLUSION: The data from this exploratory study suggest that the accumulation of molecular events seems to influence the outcome of HL, particularly in the low-IPS group. PMID- 15117990 TI - Survivin expression predicts poorer prognosis in anaplastic large-cell lymphoma. AB - PURPOSE: Survivin, a member of the inhibitor of apoptosis (IAP) family, is not detected in normal adult tissues but is overexpressed in various cancers, including some types of lymphoma. The frequency and prognostic significance of survivin expression in anaplastic large-cell lymphoma (ALCL) is unknown. MATERIALS AND METHODS: We assessed for survivin expression in 62 ALCL tumors (30 anaplastic lymphoma kinase [ALK]-positive and 32 ALK-negative) obtained before doxorubicin-based chemotherapy. Given that survivin is a target of the STAT3 signaling pathway and STAT3 is activated in ALCL, survivin expression was also correlated with STAT3 activation. RESULTS: Survivin was expressed in 34 tumors (55%) and did not correlate with ALK. A significant association between survivin expression and STAT3 activation was observed (P =.007, Fisher's exact test). For the ALK-positive group, the 5-year failure-free survival (FFS) was 34% for patients with survivin-positive ALCL compared with 100% for patients with survivin-negative ALCL (P =.009, log-rank test). For the ALK-negative group, the 5-year FFS was 46% for patients with survivin-positive tumors compared with 89% for patients with survivin-negative tumors (P =.03, log-rank test). Overall survival was similarly worse for patients with survivin-positive tumors in both the ALK-positive and ALK-negative groups. Furthermore, multivariate analysis confirmed the independent prognostic value of survivin expression, along with age older than 60 years and Ann Arbor stage III or IV. CONCLUSION: Survivin is expressed in approximately half of ALCL tumors and independently predicts unfavorable clinical outcome. Modulation of survivin expression or function may provide a novel target for experimental therapy in patients with ALCL. PMID- 15117991 TI - Localized pelvic neuroblastoma: excellent survival and low morbidity with tailored therapy--the 10-year experience of the French Society of Pediatric Oncology. AB - PURPOSE: To assess the results and morbidity of treatment of children with localized pelvic neuroblastoma (NB). PATIENTS AND METHODS: All consecutive cases of localized pelvic NB registered in the French multicenter prospective studies NBL90 and NBL94 between 1990 and 1999 were reviewed. Resectability was decided on the basis of clinical and radiologic evaluation. In unresectable tumors, primary chemotherapy (combinations of carboplatin-etoposide and vincristine cyclophosphamide-doxorubicine) was administered before surgery. RESULTS: Forty seven children (with 26 resectable tumors and 21 unresectable) were included in this study. At the end of treatment, 31 children were in complete remission (66%). Long-term neurologic sequelae were observed in seven patients (15%), directly attributable to surgery in three cases. After a median follow-up of 48 months (range, 13 to 129 months), 44 patients are alive. Six children experienced local relapse; four of these children achieved subsequent remission. The projected overall survival and event-free survival (EFS) rates at 5 years are, respectively, 93% +/- 4% and 84% +/- 5%. Survival of children treated with preoperative chemotherapy are similar to those treated by primary surgery (80% and 88% respectively). The extent of surgical resection seemed to have no influence on the outcome (EFS rates 76% and 89% in case of gross residue and complete resection or microscopic residue, respectively). CONCLUSION: Our data confirm the excellent survival of localized pelvic NBs. Considering the efficacy of preoperative chemotherapy, patients with pelvic NB should be carefully screened for primary surgery. The risk of neurologic impairment during radical excision should be balanced with the good survival of children with minimal residual disease. PMID- 15117992 TI - Increasing mixed chimerism is an important prognostic factor for unfavorable outcome in children with acute lymphoblastic leukemia after allogeneic stem-cell transplantation: possible role for pre-emptive immunotherapy? AB - PURPOSE: We recently reported that children with acute leukemias who show increasing mixed chimerism (MC) after allogeneic stem-cell transplantation have a significantly enhanced risk of relapse. Here we present the results of a prospective multicenter study to investigate (1) whether relapse of acute lymphoblastic leukemia (ALL) can be determined in advance by serial analysis of chimerism, and (2) if outcome can be influenced by withdrawal of immunosuppression and/or by low-dose donor lymphocyte infusion when increasing MC is detected. PATIENTS AND METHODS: Serial and quantitative analysis of chimerism was performed using a fluorescent-based short-tandem-repeat-polymerase chain reaction in 163 children with ALL. RESULTS: One hundred one patients revealed complete chimerism (CC) or low-level MC (CC/low-level MC); increasing MC was found in 46 patients; and decreasing MC, in 16 patients. Relapse was significantly more frequent in patients with increasing MC (26 of 46) than in patients with CC/low-level MC (eight of 101) or in patients with decreasing MC (0 of 16; P <.0001). The probability of 3-year event-free survival (EFS) was 54% for all patients, 66% for patients with CC/low-level MC (n = 101), 66% for patients with decreasing MC (n = 16), and 23% for patients with increasing MC (n = 46; P <.0001). Of the 46 patients with increasing MC, 31 received immunotherapy. This group had a significantly higher 3-year EFS estimate (37%) than the 15 patients who did not receive immunotherapy (0%; P <.001). CONCLUSION: Serial analysis of chimerism reliably identifies patients at highest risk to relapse. The 3-year EFS of patients with increasing MC without immunotherapy was 0%, by which overt relapse could be prevented in a considerable group of patients. PMID- 15117993 TI - Laboratory and clinical evidence of synergistic cytotoxicity of sequential treatment with gemcitabine followed by docetaxel in the treatment of sarcoma. AB - PURPOSE: A recent report of the combination of gemcitabine and docetaxel described favorable results in patients with uterine leiomyosarcoma. The objective of this report is to describe experience with this combination in a variety of histologic subtypes of sarcoma. Additionally, cell-culture studies were performed to assess the effect of the sequence of drug administration on colony formation. PATIENTS AND METHODS: A medical record review of 35 patients receiving the gemcitabine/docetaxel combination was undertaken. Gemcitabine 675 mg/m(2) intravenously was administered over 90 minutes on days 1 and 8, and docetaxel 100 mg/m(2) intravenously was administered over 60 minutes on day 8 of a 21-day cycle. Cell culture studies using the SAOS-2 osteosarcoma cell line and MCF-7 breast cancer cell line were also performed. Gemcitabine and docetaxel were added to cells either simultaneously for 24 hours, gemcitabine for 24 hours followed by docetaxel for 24 hours, or the reverse sequence. RESULTS: Thirty-five patients were treated. Five complete responses and 10 partial responses were observed for an overall response rate of 43%. Responses occurred in uterine, extremity, and retroperitoneal leiomyosarcoma, osteosarcomas, angiosarcomas, malignant fibrous histiocytomas, malignant peripheral-nerve sheath tumors, and Ewing's sarcoma. In the cell culture studies, gemcitabine followed by docetaxel provided synergy. In contrast, the administration of drugs simultaneously resulted in antagonism, and docetaxel followed by gemcitabine provided mixed results. CONCLUSION: The combination of gemcitabine and docetaxel seems to be active in a variety of sarcomas. A multicenter, randomized clinical trial in soft tissue sarcoma comparing gemcitabine alone with this combination, is ongoing. PMID- 15117994 TI - Randomized clinical trial of the effectiveness of a self-care intervention to improve cancer pain management. AB - PURPOSE: This randomized clinical trial tested the effectiveness of the PRO-SELF Pain Control Program compared with standard care in decreasing pain intensity scores, increasing appropriate analgesic prescriptions, and increasing analgesic intake in oncology outpatients with pain from bone metastasis. PATIENTS AND METHODS: Patients were randomly assigned to the PRO-SELF intervention (n = 93) or standard care (n = 81). Patients in the standard care arm were seen by a research nurse three times and were called three times by phone between the home visits. PRO-SELF group patients were seen by specially trained intervention nurses and received a psychoeducational intervention, were taught how to use a pillbox, and were given written instructions on how to communicate with their physician about unrelieved pain and the need for changes in their analgesic prescriptions. Patients were coached during two follow-up home visits and three phone calls on how to improve their cancer pain management. RESULTS: Pain intensity scores decreased significantly from baseline (all P <.0001) in the PRO-SELF group (ie, least pain, 28.4%; average pain, 32.5%; and worst pain, 27.0%) compared with the standard care group (ie, least increased by 14.6%, average increased by 1.9%, and worst decreased by 1.2%). The percentage of patients in the PRO-SELF group with the most appropriate type of analgesic prescription increased significantly from 28.3% to 37.0% (P =.008) compared with a change from 29.6% to 32.5% in the standard care group. CONCLUSION: The use of a psychoeducational intervention that incorporates nurse coaching within the framework of self-care can improve the management of cancer pain. PMID- 15117995 TI - Cancer patient preferences for communication of prognosis in the metastatic setting. AB - PURPOSE: To identify preferences for and predictors of prognostic information among patients with incurable metastatic cancer. PATIENTS AND METHODS: One hundred twenty-six metastatic cancer patients seeing 30 oncologists at 12 outpatient clinics in New South Wales, Australia, participated in the study. Patients were diagnosed with incurable metastatic disease within 6 weeks to 6 months of recruitment. Patients completed a survey eliciting their preferences for prognostic information, including type, quantity, mode, and timing of presentation; anxiety and depression levels; and information and involvement preferences. RESULTS: More than 95% of patients wanted information about side effects, symptoms, and treatment options. The majority wanted to know longest survival time with treatment (85%), 5-year survival rates (80%), and average survival (81%). Words and numbers were preferred over pie charts or graphs. Fifty nine percent (59%) wanted to discuss expected survival when first diagnosed with metastatic disease. Thirty-eight percent and 44% wanted to negotiate when expected survival and dying, respectively, were discussed. Patients with higher depression scores were more likely to want to know shortest time to live without treatment (P =.047) and average survival (P =.049). Lower depression levels were significantly associated with never wanting to discuss expected survival (P =.03). Patients with an expected survival of years were more likely to want to discuss life expectancy when first diagnosed with metastases (P =.02). CONCLUSION: Most metastatic cancer patients want detailed prognostic information but prefer to negotiate the extent, format, and timing of the information they receive from their oncologists. PMID- 15117996 TI - Acupuncture for postchemotherapy fatigue: a phase II study. AB - PURPOSE: To determine whether improvement in postchemotherapy fatigue following acupuncture treatment is substantial enough to warrant a controlled trial. PATIENTS AND METHODS: We accrued patients at Memorial Sloan-Kettering Cancer Center who had completed cytotoxic chemotherapy but experienced persisting fatigue. Patients with severe anemia, clinical depression, or Karnofsky performance status score less than 70 were excluded. Thirty-seven patients were registered in two cohorts; 31 provided follow-up data. Patients received acupuncture either twice per week for 4 weeks (25 patients) or once per week for 6 weeks (12 patients). The primary end point was change in score on the Brief Fatigue Inventory between baseline and 2 weeks after the final treatment. A baseline Brief Fatigue Inventory score of four or greater was an eligibility requirement for the trial. RESULTS: Patients had completed cytotoxic chemotherapy an average of more than 2 years previously. Baseline fatigue scores were high, with approximately half of the sample scoring in the "severe" range. Mean improvement following acupuncture was 31.1% (95% CI, 20.6% to 41.5%), meeting our prespecified criterion for declaring acupuncture worthy of further study. Increasing age was associated with poorer response and failure to complete the study. There was no important difference in improvement following once-weekly and twice-weekly treatments. CONCLUSION: Acupuncture is worthy of further study in the treatment of postchemotherapy fatigue. PMID- 15117997 TI - Individuals with an increased risk of colorectal cancer: perceived benefits and psychological aspects of surveillance by means of regular colonoscopies. AB - PURPOSE: To evaluate the psychological consequences of genetic counseling followed by a surveillance program using colonoscopy among individuals with increased risk of colorectal cancer. PATIENTS AND METHODS: Two hundred sixty-five individuals, participating in a surveillance program with colonoscopy, were mailed a survey questionnaire that assessed their experience of the surveillance program and their perception of the risk of colorectal cancer. The Hospital Anxiety and Depression scale and the Swedish Short Form-36 Health Survey was also included. RESULTS: Two hundred forty individuals completed the questionnaire and were divided into the following risk groups: risk group 1, an individual with a mutation in hMLH1 or hMSH2 and a lifetime colorectal cancer risk of 80% (n = 28); risk group 2, a lifetime colorectal cancer risk of 40% (n = 129); and risk group 3, a lifetime colorectal cancer risk of 20% (n = 83). Among all individuals, the mean for perceived benefit was 8.0, and the perception of discomfort was 3.3 on the visual analog scale (1-10). In risk group 1, 61% underestimated personal risks as being 40% or less. Approximately 50% of the subjects in risk groups 2 and 3 either under- or overestimated their lifetime risk. According to the Swedish Short Form-36 Health Survey and the Hospital Anxiety and Depression scale, the study sample resembled the reference population. CONCLUSION: A majority of the study sample understood why they were under surveillance, and regular colonoscopies were well-tolerated. The wide range of risk perception as well as low-risk perception in mutation positive subjects is acceptable, as long as these individuals adhere to surveillance programs and do not demonstrate increased levels of anxiety or depression. PMID- 15117998 TI - The expanding role of systemic therapy in head and neck cancer. AB - Treatment of squamous cell carcinoma of the head and neck (SCCHN) has evolved greatly in the last two decades, owing to the integration of chemotherapy, advances in radiotherapeutic techniques, and organ-preserving surgery. Several randomized trials have established new standards of care that should be adopted. Current efforts are building on these earlier trials in order to improve survival and quality of life. Coincident with this, investigators are developing molecularly targeted approaches that hold promise for the future. This review will focus on current therapy for locally advanced, recurrent, and metastatic SCCHN and discuss controversies and directions for future research. PMID- 15117999 TI - Germ cell tumors. Case 1. Intraocular metastases from testicular cancer. PMID- 15118000 TI - Germ cell tumors. Case 2. Unusual course of pure testicular seminoma. PMID- 15118001 TI - Germ cell tumors. Case 3. Primary yolk sac tumor of the liver. PMID- 15118002 TI - Treatment decision aids in advanced cancer: when the goal is not cure and the answer is not clear. PMID- 15118003 TI - Operate on my stomach cancer? Oh, no--not you, or not yet! PMID- 15118005 TI - The field of radiation therapy may effect health-related quality of life in patients with operable breast cancer. PMID- 15118007 TI - Is docetaxel now an essential component of neoadjuvant breast chemotherapy? PMID- 15118009 TI - Use of gabapentin in the prevention of taxane-induced arthralgias and myalgias. PMID- 15118010 TI - Time to stop progesterone receptor testing in breast cancer management. PMID- 15118012 TI - General slowing or decreased inhibition? Mathematical models of age differences in cognitive functioning. AB - Researchers have attempted to explain age-related decrements in cognitive performance in terms of reduced processing speed or decreased ability to inhibit irrelevant thoughts. We present these ideas in the context of a dynamic model derived from extensions of the classical predator-prey equation. Reduced processing speed among older adults is represented by use of delays in the dynamic model, whereas the interference imposed by distractors is captured by use of the predator-prey interaction term. We demonstrate the versatility of this modeling approach, and its pertinence to age-related behavioral change, by means of numerical simulations. In showing the applicability of these models, we identify several unresolved methodological and measurement issues that have to be addressed. PMID- 15118013 TI - Personality and mortality in old age. AB - We examined the relation of personality to mortality in 883 older Catholic clergy members (69% women). At baseline, they completed the NEO Five-Factor Inventory, which assesses the five principal dimensions of personality. They were followed for a mean of 5.1 years, during which 182 deaths occurred. Risk of death was nearly doubled in those with a high neuroticism score (90th percentile) compared with a low score (10th percentile) and was approximately halved in those with a high conscientiousness score compared with a low score. Findings for extraversion were mixed, and neither agreeableness nor openness was strongly related to mortality. The results suggest that personality is associated with mortality in old age. PMID- 15118014 TI - The complexity of emotions in later life. AB - The complexity of positive and negative emotions was examined in a sample of 40 adults between the ages of 60 and 85 years. Participants' emotional experiences were assessed by use of a 30-day assessment protocol. Results suggest that different vulnerability and resilience factors are implicated in the intraindividual experience of positive and negative emotions. Individual differences in perceived stress and neuroticism were associated with less differentiation and fewer co-occurrences of positive and negative emotional experiences. In contrast, dispositional resilience predicted greater differentiation and more co-occurrences of affective states. Findings are interpreted within the framework of life-span theories of emotions. PMID- 15118015 TI - Spirituality and well-being in frail and nonfrail older adults. AB - Previous studies have identified that spiritual beliefs contribute to psychological well-being (PWB) in older people, but limited research has considered the effects of spirituality on PWB when physical health deteriorates and people become frail. We recruited 233 British participants from warden controlled retirement housing to complete interviewer-administered questionnaires. Results showed that, after we controlled for marital status, age, education, other health problems, and gender, degree of frailty had a negative effect on PWB. Spirituality was also a significant predictor of PWB and moderated the negative effects of frailty on PWB. Therefore, this study suggests that spirituality is a resource in maintaining PWB, and that the use of this resource is more significant for individuals with greater levels of frailty. PMID- 15118016 TI - The extent of stability and change in episodic and semantic memory in old age: demographic predictors of level and change. AB - Structural stability and change in semantic and episodic memory performance as well as interindividual differences in 5-year changes in these constructs are examined within a sample of older adults (age rangeT1 = 60-80; n = 361). Interindividual differences in change were limited but significant. Stability coefficients were higher for semantic memory (.95) than for episodic memory (.87). Changes in episodic and semantic memory performance were strongly associated (r =.68). Across time, variances and covariances increased, and a tendency toward dedifferentiation in terms of increasing correlations was found. Chronological age was related to both level and change, but gender and education were only related to level of memory performance. Collectively, these results depict relatively high degrees of structural stability and stability of interindividual differences in declarative memory in old age. PMID- 15118017 TI - Age-related differences in arithmetic problem-verification strategies. AB - To test age-related differences in split and problem-difficulty effects, adults between the ages of 20 and 80 years (N = 138) performed a simple and a complex inequality verification task (e.g., 6 + 3 < 11, 271 + 182 < 458; true or false?). Split effects in verification tasks (i.e., better performance for large-split than for small-split problems) reflect strategy selection between nonexhaustive verification (e.g., evaluation of plausibility; estimation) and exhaustive verification (e.g., retrieval; calculation). Problem-difficulty effects (i.e., better performance for easy than hard problems) reflect calculation processing. Results showed decreased split effects across age groups, particularly in the complex task. Moreover, problem-difficulty effects did not vary across age groups. Age-related changes were mostly mediated by age-related declines in processing speed. PMID- 15118018 TI - Decline in material family support for older people in urban Ghana, Africa: understanding processes and causes of change. AB - OBJECTIVES: Material family support for older people in Ghana, as in other African countries, has declined in recent decades, exposing increasing numbers especially of urban elderly to destitution and poverty. The nature and causes of this decline remain poorly understood, in particular the relative role of growing material constraints, as proposed by political economy perspectives, or weakening traditional values, as suggested by modernization perspectives. This article develops an interpretively grounded understanding of the processes underpinning the decline specifically in Accra, Ghana's capital. METHODS: A qualitative investigation of the perspectives of a three-generational respondent sample, spanning major income, ethnic, and gender groups, was conducted. RESULTS: The decline has been underpinned by two major shifts: (a) a declining resource capacity of the young to provide support and (b) a shift in the basis of filial support toward an increasing dependence on parents' past conduct and the principle of reciprocity. Normative expectations emphasizing self-reliance in old age are emerging as a result of the decline. DISCUSSION: The shifts have been caused by a complex interaction between growing resource constraints and changing values not captured by existing accounts. The dominant factor driving the change in support norms and patterns has been the change in families' material circumstances. PMID- 15118019 TI - Can culture help explain the physical health effects of caregiving over time among African American caregivers? AB - OBJECTIVES: The purposes of this study were to longitudinally examine the health outcomes of 107 African American caregivers who provided care to their elderly dependent family members and to determine the role of culture in predicting health outcomes. METHODS: With use of the stress and coping model of Pearlin and colleagues (1990) as a guide, the direct effects of background characteristics and stressors and the direct and mediating effects of resources (including culture) on two caregiver health outcomes (i.e., psychosocial health and physical functioning) were analyzed with hierarchical multiple regression analyses. RESULTS: Similar to other studies, we found that combinations of caregiver background characteristics, stressors, and resources at wave 1 had direct effects on African American caregivers' health outcomes at wave 3. Unlike previous studies, where culture was not measured, we found that cultural beliefs and values did help to explain health outcomes for African American caregivers. Specifically, culture justifications for caregiving, baseline psychosocial health, and caregiving mastery predicted wave 3 psychosocial health. Caregiver education, number of morbidities, and physical functioning at wave 1 were associated with physical functioning at wave 3. DISCUSSION: The findings from this study have implications for future studies, particularly in regard to cultural beliefs and values among African American caregivers. PMID- 15118020 TI - A longitudinal study of black-white differences in social resources. AB - OBJECTIVES: This study examined the differences in levels of social networks and social engagement between older Blacks and Whites and racial differences in rates of change in social resources over time. METHODS: The sample consisted of 5,102 Blacks and Whites, aged 65 and older, from the Chicago Health and Aging Project, who underwent up to three interviews during an average of 5.3 years of follow-up. Repeated measures of social networks were based on the number of children, relatives, and friends seen at least once a month. Social engagement was measured at each interview with four items related to social and productive activity. Random effects models were fitted to test the effect of race on social resources and change in resources over time. RESULTS: Blacks had smaller social networks and lower levels of social engagement than Whites. Racial differences in these resources were stable during follow-up. Socioeconomic status reduced racial differences in social engagement but not social networks. DISCUSSION: The findings provide evidence for lower levels of social resources among older Blacks. There was little evidence that racial differences were the result of greater decline during old age itself. PMID- 15118021 TI - Work at older ages in Japan: variation by gender and employment status. AB - OBJECTIVES: This study describes the correlates of labor force participation among Japanese men and women aged 60-85 and examines differences by gender and employment status. METHODS: Using four waves of data collected from a national sample of older Japanese between 1990 and 1999, we estimate multinomial logistic regression models for three measures of labor force participation (current labor force status, labor force exit, and labor force re-entry) as a function of individual and family characteristics measured 3 years earlier. RESULTS: Labor force participation is significantly associated with socioeconomic status, longest occupation, and family structure. The strength and nature of these relationships differ markedly for men and women and for wage employment and self employment. DISCUSSION: The emphasis on life course experiences and work-family interdependence characterizing recent research on retirement in the United States is clearly relevant in Japan as well. To better understand later-life labor force participation in Japan, subsequent research should incorporate more direct measures of life course experiences and family relationships and attempt to make explicit cross-national comparisons of these relationships. PMID- 15118022 TI - The effects of residential mobility on ADL and IADL limitations among the very old living in the community. AB - OBJECTIVES: We attempt to understand the influence of residential mobility on changes in objective activity of daily living (ADL) and instrumental ADL (IADL) limitations, while taking into account the subjective reason for the move. METHODS: We examine noninstitutionalized adults aged 70 and older who are in the Longitudinal Study of Aging. We use bivariate regression to identify differences between nonmovers and various types of movers in 1988 and 1990 ADL and IADL limitations. Multivariate residualized regression models estimate the effect of residential mobility on the changes in limitations between 1984-1988 and 1988 1990. We give particular attention to the timing and reason for the move. RESULTS: Controlling for demographic, social support, health status, and social integration characteristics, we find that residential mobility is associated with a short-term increase in ADL and IADL limitations. However, an additional analysis reveals that this short-term increase in limitations is only experienced by older adults who move for subjective health reasons. Compared with nonmovers, movers do not have significantly different changes in limitations over the long term. DISCUSSION: Older adults and service providers need to be concerned with short-term increases in objective ADL and IADL limitations that accompany residential relocations, especially for those who move for subjective health reasons. However, from a long-term perspective, residential relocation may serve as a mechanism for accommodating age-related changes that threaten effective functioning. PMID- 15118023 TI - Depression among recipients of informal care: the effects of reciprocity, respect, and adequacy of support. AB - OBJECTIVES: The objective of this work was to examine the relationship of perceived quality of care to depression among recipients of informal long-term care. METHODS: Generalized estimating equations were used to generate population average logistic regression models of prevalent depression, using a sample of 420 disabled community-dwelling women aged 65 or older receiving informal care obtained from the Women's Health and Aging Study Caregiving Survey. RESULTS: Findings confirm a substantial prevalence of depression among older women with disabilities and support the hypothesis that perceived reciprocity and respect afforded by one's primary caregiver as well as adequacy of instrumental support all were associated with a lower likelihood of being categorized as depressed, even after controlling for sociodemographic, health, and psychosocial characteristics that are known to be related to depression. DISCUSSION: Perceived quality of informal care arrangements has a bearing on the psychological health of care recipients. Individuals in more reciprocal relationships and in relationships where they felt respected and valued were less likely to be depressed than their counterparts. PMID- 15118024 TI - The effect of the duration of follow-up in mortality analysis: the temporal pattern of different predictors. AB - OBJECTIVES: This study presents a model of the mechanisms affecting how time since baseline affects the correlation between mortality and commonly used predictors. METHODS: In 1986, 421 persons (aged 75 years or older) in a Swedish community were interviewed. Fifteen-year mortality rates were analyzed by using hazard regressions. Rather than using average risk over the whole follow-up time, this study looks at temporal differences in predictor strength. RESULTS: All studied health variables, living conditions, and life satisfaction were much stronger predictors of mortality during the first 1 or 2 years of follow-up than during later years. Gender, social contacts, and mental status were about equally correlated to mortality throughout the period. DISCUSSION: Of the presented mechanisms affecting predictive strength, results suggest the importance of the instability of predictors over time. Especially in old populations, predictors that can change rapidly (e.g., health) are strongest for the short term, revealing a lower average mortality risk for longer follow-ups. Rather stable variables (e.g., gender or social contacts) are not affected by the length of follow-up. When average risk is studied over a longer follow-up, insignificant results may hide significant effects during a part of the follow-up. These findings are relevant for studies that examine any kind of outcome after a follow up. PMID- 15118025 TI - Impact of self-assessed hearing loss on a spouse: a longitudinal analysis of couples. AB - OBJECTIVES: Hearing loss is increasingly common among older persons and is negatively associated with health and well-being. Its impact on spouses, however, is poorly researched. This study analyzed the relationship between a spouse's self-assessed hearing loss and his or her partner's physical, psychological, and social well-being 5 years later. METHODS: Subjects were 418 older married couples from the Alameda County Study. Hearing loss and adjustment variables were assessed in 1994 and outcomes in 1999. Longitudinal analyses included multivariate statistical models using generalized estimating equations to adjust for paired data and partners' hearing loss, age, gender, chronic conditions, and financial problems. RESULTS: Spouse hearing loss increased the likelihood of subsequent poorer physical, psychological, and social well-being in partners. The negative impact of husbands' hearing loss on wives' well-being appears stronger than the reverse. DISCUSSION: Findings suggest that early diagnosis and treatment of hearing loss constitute important clinical strategies to enhance the well being of both hearing-impaired individuals and their spouses and support policy change to cover hearing devices by insurance. Further research incorporating theoretical perspectives from communication theory and qualitative methodology would enhance understanding of how hearing loss impacts older couples and support refinement of interventions to promote quality of life. PMID- 15118026 TI - Longevity and health care expenditures: the real reasons older people spend more. PMID- 15118028 TI - Low-back pain following surgery for lumbar disc herniation. A prospective study. AB - BACKGROUND: Lumbar disc herniation often causes sciatica. Although surgery may provide relief of sciatic pain, it is uncertain how surgery affects the relief of low-back pain. The purpose of the present prospective study was to assess the efficacy of discectomy in the treatment of low-back pain associated with lumbar disc herniation. METHODS: Between 1998 and 2001, forty consecutive patients with single-level, unilateral lumbar disc herniation were treated surgically. The first twenty patients (Group 1) underwent standard discectomy, and the second twenty (Group 2) underwent microendoscopic discectomy. Curettage of the disc space was not performed. All forty patients were prospectively followed, and clinical outcomes were evaluated with use of a questionnaire. The mean duration of follow-up was forty months. RESULTS: All forty patients were satisfied with the outcome. Leg pain decreased rapidly (within one month) in all patients and continued to decrease at the time of the latest follow-up. There was no significant difference between the two groups in terms of leg pain, with the numbers available (p = 0.39). A significant decrease in the mean low-back pain score was noted at the time of the latest follow-up (p = 0.0007). CONCLUSIONS: Excision of a herniated disc for relief of sciatica provided rapid relief of sciatica and low-back pain. The findings of the present small study suggest that lumbar disc herniation might be a possible cause of low-back pain. PMID- 15118029 TI - The fate of patients not returning for follow-up five years after total knee arthroplasty. AB - BACKGROUND: Patients who do not return for follow-up after total knee arthroplasty are often assumed to have had a worse outcome. We postulated that the Internet may be useful for locating patients who do not return for follow-up. The purpose of this study was to compare outcomes between patients who attended prescribed postoperative appointments and those who did not. METHODS: We retrospectively reviewed the outcomes for 161 patients (200 knees) at a minimum of five years after a total knee replacement. All patients who had had no contact of any type with their surgeon in any manner beyond six months following the date of the surgery were classified as not having returned for follow-up. When patients could not be located with use of all available information in their chart, a standardized Internet search algorithm was employed with use of readily available and free search engines. RESULTS: Thirty patients with thirty-five treated knees met the definition of not returning for follow-up. Thirteen patients could not be located with use of the contact information given at their last visit. All were located with use of the Internet. None of the patients had had knee surgery elsewhere. The reasons for not adhering to the prescribed postoperative regimen were identified. The group had improved Knee Society pain and function scores compared with the preoperative values, and the patients were not significantly different from those who did return for follow-up. CONCLUSIONS: The patients who did not attend follow-up appointments in this series had not had any additional surgery and did not have any significant differences in measured outcome variables when compared with patients who had complied with a follow-up protocol. The Internet proved to be a valuable tool for locating patients who had not returned for follow-up postoperatively. PMID- 15118030 TI - American Academy of Orthopaedic Surgeons lower limb outcomes assessment instruments. Reliability, validity, and sensitivity to change. AB - BACKGROUND: The American Academy of Orthopaedic Surgeons (AAOS) has developed an array of outcomes assessment instruments designed for the efficient collection of outcomes data from patients of all ages with musculoskeletal conditions in all body regions. The Lower Limb Instruments were developed through a process of literature review, consensus-building, and field-testing. METHODS: The instruments were distributed to a total of 290 subjects in twenty orthopaedic practices throughout the United States and Canada. Of the 290 patients, seventy each had a diagnosis in the categories of foot and ankle, sports/knee, and hip and knee and forty each had a diagnosis in the categories of trauma and rehabilitation. Retests to be taken twenty-four hours after the first test were distributed to subsamples of patients for each instrument. Seventy-one one-year follow-up questionnaires (twenty-five Sports/Knee, twenty-five Foot and Ankle, sixteen Hip and Knee, and five Lower Limb Core instruments) were returned. RESULTS: The Lower Limb Core Scale and the Hip and Knee Core Scale, each consisting of seven items addressing pain, stiffness and swelling, and function, performed at an acceptable level. Additional Sports/Knee and Foot and Ankle Modules proved to have internal and retest reliability of 0.80 or better, comparable with the values for well-established measures such as the Short Form 36 (SF-36). All of the new scales were moderately to strongly correlated with other measures of pain and function, such as physician ratings, the SF-36, and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC). Seventy-one patients provided follow-up information for the analysis of sensitivity to change. The Lower Limb Core was found to contribute independently to the prediction of the transition score based on the patient and physician assessments of change. CONCLUSIONS: The AAOS Lower Limb Instruments for outcomes assessment are highly reliable and are correlated with other measures for similar constructs. They are also sensitive to change in patient status. The Lower Limb Core Scale may be used with attribution of pain either to the lower limb or to a specific joint or side without sacrificing reliability. Combined with the SF-36, the AAOS outcomes assessment instruments comprehensively and efficiently measure outcomes in orthopaedic patients with lower-limb conditions. PMID- 15118031 TI - Vascular injuries in knee dislocations: the role of physical examination in determining the need for arteriography. AB - BACKGROUND: Popliteal artery injury is frequently associated with knee dislocation following blunt trauma, an injury that is being seen with increasing frequency. The primary purpose of the present study was to evaluate the use of physical examination to determine the need for arteriography in a large series of patients with knee dislocation. The secondary purpose was to evaluate the correlation between physical examination findings and clinically important vascular injury in the subgroup of patients who underwent arteriography. METHODS: One hundred and thirty consecutive patients (138 knees) who had sustained an acute multiligamentous knee injury were evaluated at our level-1 trauma center between August 1996 and May 2002 and were included in a prospective outcome study. Four patients (four knees) were lost to follow-up, leaving 126 patients (134 knees) available for inclusion in the study. The results of the physical examination of the vascular status of the extremities were used to determine the need for arteriography. The mean duration of follow-up was nineteen months (range, eight to forty-eight months). Physical examination findings, magnetic resonance imaging findings, and surgical findings were combined to determine the extent of ligamentous damage. RESULTS: Nine patients had flow-limiting popliteal artery damage, for an overall prevalence of 7%. Ten patients had abnormal findings on physical examination, with one patient having a false-positive result and nine having a true-positive result. The knee dislocations in the nine patients with popliteal artery damage were classified, according to the Wascher modification of the Schenck system, as KD-III (one knee), KD-IV (seven knees), and KD-V (one knee). CONCLUSIONS: Selective arteriography based on serial physical examinations is a safe and prudent policy following knee dislocation. There is a strong correlation between the results of physical examination and the need for arteriography. Increased vigilance may be justified in the case of a patient with a KD-IV dislocation, for whom serial examinations should continue for at least forty-eight hours. PMID- 15118032 TI - Topical glyceryl trinitrate treatment of chronic noninsertional achilles tendinopathy. A randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: Noninsertional Achilles tendinopathy is a degenerative overuse disorder. No method has been universally successful in treating this condition. Topically applied nitric oxide has been shown, in animal models, to be effective for the treatment of fractures and cutaneous wounds through mechanisms that may include stimulation of collagen synthesis in fibroblasts. The goal of the present study was to determine if topical glyceryl trinitrate improves clinical outcome measures in patients with Achilles tendinopathy. METHODS: A prospective, randomized, double-blind, placebo-controlled trial involving a total of sixty five patients (eighty-four Achilles tendons) was performed to compare continuous application of topical glyceryl trinitrate (at a dosage of 1.25 mg per twenty four hours) with rehabilitation alone for the treatment of noninsertional Achilles tendinopathy. RESULTS: Compared with the control group, the glyceryl trinitrate group showed reduced pain with activity at twelve weeks (p = 0.02) and twenty-four weeks (p = 0.03), reduced night pain at twelve weeks (p = 0.04), reduced tenderness at twelve weeks (p = 0.02), decreased pain scores after the hop test at twenty-four weeks (p = 0.005), and increased ankle plantar flexor mean total work compared with the baseline level at twenty-four weeks (p = 0.04). Twenty-eight (78%) of thirty-six tendons in the glyceryl trinitrate group were asymptomatic with activities of daily living at six months, compared with twenty (49%) of forty-one tendons in the placebo group (p = 0.001, chi-square analysis). The mean effect size for all outcome measures was 0.14. CONCLUSIONS: Topical glyceryl trinitrate significantly reduced pain with activity and at night, improved functional measures, and improved outcomes in patients with Achilles tendinopathy. PMID- 15118033 TI - Demographics of traumatic amputations in children. Implications for prevention strategies. AB - BACKGROUND: The demographics of traumatic amputations in children are not well known. The purpose of this review was to identify those demographics for use as a possible guide for prevention strategies. METHODS: The study was a retrospective review of the data on children with traumatic amputation who had received care at one center in the upper midwestern United States from 1980 to 2000. The child's gender and age at the time of the amputation, the date and etiology of the amputation, and the amputation level were tabulated. Statistical analyses of seasonal variations were performed. RESULTS: There were 256 amputations in 235 children. The mean age (and standard deviation) at the time of the amputation was 7.9 +/- 5.0 years. The amputation involved one extremity in 217 children, two extremities in sixteen, and three and four extremities in one child each. Of the 256 amputations, 165 involved the lower extremity. The traumatic amputation was caused by a lawnmower in sixty-nine children, farm machinery in fifty-seven, a motor-vehicle accident in thirty-eight, a train in twenty, and miscellaneous mechanisms in fifty-one. The mean age at the time of the injury varied according to the mechanism of injury and ranged from 1.9 years for burns to 11.5 years for boating injuries. Fifty-four (78%) of the sixty-nine children with a lawnmower amputation were five years of age or less. There were significant seasonal variations: the mean date of the lawnmower injuries was June 10, the mean date of the farming injuries was September 2, and the mean date of the motor-vehicle related injuries was July 16. CONCLUSIONS: There are common patterns of traumatic amputations in children based on the mechanism of injury, the season, and the age of the child. The ideal time for an educational campaign for the prevention of lawnmower injuries appears to be March and April and should be directed toward parents. The best times for such a campaign for the prevention of farming-related accidents appear to be both the spring and the early fall, and the campaign should be directed toward both parents and older children. PMID- 15118034 TI - Cementing a liner into a stable cementless acetabular shell: the double-socket technique. AB - BACKGROUND: During revision hip replacement surgery, the cementless acetabular shell is often well fixed but the locking mechanism may be ineffective. Cementing a new liner into the existing acetabular shell (the double-socket technique) can provide a simple solution. The purposes of the present study were to review our initial clinical results and to define the potential limitations of this technique. METHODS: Thirty-two hips with a preexisting well-fixed acetabular socket that had been in situ for an average of 8.6 years were treated with the insertion of a new polyethylene liner (seventeen hips) or a metal liner (fifteen hips) with use of cement. The indication for this technique was a deficient locking mechanism in twenty-two hips and the unavailability of a matching liner in ten hips. Anteroposterior radiographs of all hips were analyzed by a single independent reviewer. RESULTS: The mean duration of follow-up was 5.1 years. Six hips required a reoperation after a mean of 29.7 months; the reasons for the reoperations included aseptic failure of the acetabular construct (four hips), instability (one hip), and sepsis (one hip). The University of California at Los Angeles hip scores improved significantly (p < 0.001) compared with the preoperative values; specifically, the mean score improved from 6.2 to 9.1 for pain, from 6.3 to 8.3 for walking, from 6.2 to 7.8 for function, and from 4.7 to 5.8 for activity. The prevalence of dislocation was 22%. Kaplan-Meier analysis with revision as the end point revealed a five-year survival rate of 78% (95% confidence interval, 55% to 91%). CONCLUSIONS: The double-socket technique is a good alternative to acetabular socket removal for suitable candidates who have a well-fixed cementless socket with an inner diameter that is larger than the outer diameter of the cemented liner. This technique preserves acetabular bone stock and permits conversion to alternate bearing surfaces. We believe, however, that removal of a well-fixed acetabular shell or the use of a constrained liner should be strongly considered for patients with a history of hip instability. PMID- 15118035 TI - Long-term changes of the nonresurfaced patella after total knee arthroplasty. AB - BACKGROUND: The most common complications of total knee arthroplasty involve the patellofemoral joint. However, the long-term fate of the nonresurfaced patella after total knee arthroplasty has seldom been reported. The purpose of this study was to evaluate the long-term changes of the nonresurfaced patella after total knee arthroplasty. METHODS: We retrospectively evaluated the nonresurfaced patella in 227 knees (181 patients) at an average of 8.5 years after total knee arthroplasty. Functional results were correlated with radiographic changes, and risk factors leading to abnormal changes of the patella were analyzed. RESULTS: Preoperatively, 186 (82%) of the patellae tracked centrally, thirty-nine (17%) displaced or tilted laterally, and two (1%) subluxated laterally. At the time of the latest follow-up, 133 (59%) of the patellae still tracked centrally with preservation of the cartilage thickness, fifteen (7%) showed early lateral tilt, sixty-eight (30%) had progressive loss of lateral cartilage thickness with lateral tilt and/or displacement, seven (3%) had progressed to lateral subluxation, and four (2%) tilted medially. An abnormal patellofemoral joint did not affect the knee and functional scores (p = 0.90 and 0.89, respectively). However, symptoms such as difficulty rising from a chair or reluctance to use the involved lower limb while climbing stairs were noted. Preoperative patellar maltracking was identified as the only risk factor leading to postoperative patellar abnormalities (relative risk, 2.7; 95% confidence interval, 2.21 to 3.30; p = 0.003). CONCLUSIONS: At the time of follow-up, at an average of 8.5 years, patellar tracking and the patellofemoral joint remained normal after approximately 60% of the total knee arthroplasties performed without resurfacing of the patella. Progressive degenerative changes of the nonresurfaced patella (mainly on the lateral facet) and patellar maltracking were the most common abnormal radiographic changes. Patients with preoperative patellar maltracking were at risk for the development of these changes and clinical symptoms. Resurfacing of the patella during total knee arthroplasty may benefit such patients. PMID- 15118036 TI - Distal humeral fractures treated with noncustom total elbow replacement. AB - BACKGROUND: The purpose of this study was to review the cases of patients with a distal humeral fracture that was treated with a noncustom total elbow arthroplasty. We hypothesized that, on the basis of the functional and clinical outcome, total elbow replacement is a reliable option for the treatment of elderly patients with a severe, comminuted fracture of the distal part of the humerus. METHODS: We retrospectively reviewed forty-nine acute distal humeral fractures in forty-eight patients who were treated with total elbow arthroplasty as the primary option. The average age of the patients was sixty-seven years. Forty-three fractures were followed for at least two years. According to the AO classification, five fractures were type A, five were type B, and thirty-three were type C. The average age of the forty-three patients was sixty-nine years and the average duration of follow-up was seven years. Fourteen patients died during the review period. Postoperative clinical function was assessed with use of the Mayo elbow performance score, and anteroposterior and lateral radiographs made at follow-up examinations were reviewed. RESULTS: At the latest follow-up examination, the average flexion arc was 24 degrees (range, 0 degrees to 75 degrees ) to 131 degrees (range, 100 degrees to 150 degrees ) and the Mayo elbow performance score averaged 93 of a possible 100 points. Heterotopic ossification was present to some extent in seven elbows, with radiographic abutment noted in two. Thirty-two (65%) of the forty-nine elbows had neither a complication nor any further surgery from the time of the index arthroplasty to the most recent follow up evaluation. Fourteen elbows (29%) had a single complication, and most of them did not require further surgery. Ten additional procedures, including five revision arthroplasties, were required in nine elbows; five were related to soft tissue and five were related to the implant or bone. CONCLUSIONS: Complex distal humeral fractures should be assessed primarily for the reliability with which they can be reconstructed with osteosynthesis. When osteosynthesis is not considered to be feasible, especially in patients who are physiologically older and place lower demands on the joint, total elbow arthroplasty can be considered. This retrospective review supports a recommendation for total elbow arthroplasty for the treatment of an acute distal humeral fracture when strict inclusion criteria are observed. PMID- 15118037 TI - Long-term results of segmental prosthesis fixation by extracortical bone-bridging and ingrowth. AB - BACKGROUND: The technique of extracortical bone-bridging and ingrowth fixation with a porous coating over the shoulder region of the implant and augmentation by autogenous bone-grafting was introduced to improve the longevity of implant fixation. The potential advantages of this technique are that new-bone formation across the bone-prosthesis junction may share stress and may prevent osteolysis by sealing off this critical region against the infiltration of wear particles. The objectives of this study were to examine the prevalence of stem-loosening with use of the extracortical bone-bridging and ingrowth technique, the amount of bone formation over the porous-coated region of this prosthesis, and the characteristics of bone formation over the porous-coated region and adjacent bone. METHODS: Forty-three patients who had prosthetic reconstruction with the extracortical bone-bridging and ingrowth technique from 1976 to 1990 were included in this retrospective study. The mean length of follow-up was 9.7 years (range, two to twenty-one years). All but one patient were managed with autogenous bone graft; five, with allograft and autograft; and one, with allograft only. Extracortical bone formation was measured over a 2-cm length of the porous-coated region of the prosthesis in four zones (the medial and lateral aspects on anteroposterior radiographs and the anterior and posterior aspects on lateral radiographs) and was reported as the percentage of the total length (8 cm) covered by extracortical bone with a thickness of >1 mm. The Spearman rank coefficient was used to assess the correlation between pairs of continuous variables. RESULTS: The final average percentage of the porous-coated region that was covered by extracortical bone formation was 76% +/- 34% for all patients and anatomical sites of reconstruction. Use of bone cement was associated with less bone formation (p = 0.04), and this value remained lower at the final measurement (p = 0.06). One stem had aseptic loosening, but no sign of osteolysis was found. The radiographic appearance of the bone formation had stabilized at two years of follow-up. All patients with allograft augmentation had greater bone formation. The amount of extracortical bone formation did not differ in relation to the type of porous coating, anatomical sites, pathological disorder, sex or age of the patient, or length of reconstruction. CONCLUSIONS: As shown by the low prevalence of stem-loosening (two of fifty-six stems or one of forty-three patients), the use of the extracortical bone-bridging and ingrowth fixation technique is associated with improved stem fixation in segmental bone-replacement prostheses applied for limb salvage. In the demanding biomechanical environment and with the risk of stress and particle-related bone resorption, the extracortical bone bridging and ingrowth fixation is an attractive method to provide long-lasting implant fixation. PMID- 15118038 TI - Differentiation between septic arthritis and transient synovitis of the hip in children with clinical prediction algorithms. AB - BACKGROUND: Differentiation between septic arthritis and transient synovitis of the hip in children can be difficult. Kocher et al. recently developed a clinical prediction algorithm for septic arthritis based on four clinical variables: history of fever, non-weight-bearing, an erythrocyte sedimentation rate of >or=40 mm/hr, and a serum white blood-cell count of >12000/mm(3) (>12.0 x 10(9)/L). The purpose of this study was to apply this clinical algorithm retrospectively to determine its predictive value in our patient population. METHODS: A retrospective review was performed to identify all children who had undergone a hip arthrocentesis for the evaluation of an irritable hip at our institution between 1992 and 2000. One hundred and sixty-three patients with 165 involved hips satisfied the criteria for inclusion in the study and were classified as having true septic arthritis (twenty hips), presumed septic arthritis (twenty seven hips), or transient synovitis (118 hips). RESULTS: Patients with septic arthritis (true and presumed; forty-seven hips) differed significantly (p < 0.05) from patients with transient synovitis (118 hips) with regard to the erythrocyte sedimentation rate, differential of serum white blood-cell count, total white blood-cell count and differential in the synovial fluid, gender, previous health care visits, and history of fever. If the four independent multivariate predictors of septic arthritis proposed by Kocher et al. were present, the predicted probability of the patient having septic arthritis was 59% in our study, in contrast to the 99.6% predicted probability in the patient population described by Kocher et al. Statistical analyses demonstrated that the best model to describe our patient population was based on three variables: a history of fever, a serum total white blood-cell count of >12000/mm(3) (>12.0 x 10(9)/L), and a previous health-care visit. When all three variables were present, the predicted probability of the patient having septic arthritis was 71%. CONCLUSIONS: Although the use of a clinical prediction algorithm to differentiate between septic arthritis and transient synovitis may have improved the utility of existing technology and medical care to facilitate the diagnosis at the institution at which the algorithm originated, application of the algorithm proposed by Kocher et al. or of our three-variable model does not appear to be valid at other institutions. PMID- 15118039 TI - Health-related quality of life in total hip and total knee arthroplasty. A qualitative and systematic review of the literature. AB - BACKGROUND: Total hip and total knee arthroplasties are well accepted as reliable and suitable surgical procedures to return patients to function. Health-related quality-of-life instruments have been used to document outcomes in order to optimize the allocation of resources. The objective of this study was to review the literature regarding the outcomes of total hip and knee arthroplasties as evaluated by health-related quality-of-life instruments. METHODS: The Medline and EMBASE medical literature databases were searched, from January 1980 to June 2003, to identify relevant studies. Studies were eligible for review if they met the following criteria: (1). the language was English or French, (2). at least one well-validated and self-reported health-related quality of life instrument was used, and (3). a prospective cohort study design was used. RESULTS: Of the seventy-four studies selected for the review, thirty-two investigated both total hip and total knee arthroplasties, twenty-six focused on total hip arthroplasty, and sixteen focused on total knee arthroplasty exclusively. The most common diagnosis was osteoarthritis. The duration of follow-up ranged from seven days to seven years, with the majority of studies describing results at six to twelve months. The Short Form-36 and the Western Ontario and McMaster University Osteoarthritis Index, the most frequently used instruments, were employed in forty and twenty-eight studies, respectively. Seventeen studies used a utility index. Overall, total hip and total knee arthroplasties were found to be quite effective in terms of improvement in health-related quality-of-life dimensions, with the occasional exception of the social dimension. Age was not found to be an obstacle to effective surgery, and men seemed to benefit more from the intervention than did women. When improvement was found to be modest, the role of comorbidities was highlighted. Total hip arthroplasty appears to return patients to function to a greater extent than do knee procedures, and primary surgery offers greater improvement than does revision. Patients who had poorer preoperative health-related quality of life were more likely to experience greater improvement. CONCLUSIONS: Health-related quality-of-life data are valuable, can provide relevant health-status information to health professionals, and should be used as a rationale for the implementation of the most adequate standard of care. Additional knowledge and scientific dissemination of surgery outcomes should help to ensure better management of patients undergoing total hip or total knee arthroplasty and to optimize the use of these procedures. PMID- 15118040 TI - Coronoid process and radial head as posterolateral rotatory stabilizers of the elbow. AB - BACKGROUND: The purpose of this study was to evaluate the role of the radial head and the coronoid process as posterolateral rotatory stabilizers of the elbow and to determine the stabilizing effect of radial head replacement and coronoid reconstruction. METHODS: The posterolateral rotatory displacement of the ulna was measured after application of a valgus and supinating torque (1). in seven intact elbows, (2). after radial head excision, (3). after sequential resection of the coronoid process, (4). after subsequent insertion of each of two different types of metal radial head prostheses (a rigid implant and a bipolar implant with a floating cup), and (5). after subsequent reconstruction of the coronoid with each of two different techniques in the same cadaveric elbow. RESULTS: The posterolateral rotatory laxity averaged 5.4 degrees in the intact elbows. The surgical approach used in this study insignificantly increased the mean laxity to 9 degrees. Excision of the radial head in an elbow with intact collateral ligaments caused a mean posterolateral rotatory laxity of 18.6 degrees (p < 0.0001). Additional removal of 30% of the height of the coronoid fully destabilized the elbows, always resulting in ulnohumeral dislocation despite intact ligaments. Implantation of a rigid radial head prosthesis stabilized the elbows. However, a mean laxity of 16.9 degrees persisted after insertion of a floating prosthesis (p < 0.0001). The elbows with a defect of 50% or 70% of the coronoid, loss of the radial head, and intact ligaments could not be stabilized by radial head replacement alone, but additional coronoid reconstruction restored stability. CONCLUSIONS: The results of this study suggest that the coronoid and the radial head contribute significantly to posterolateral rotatory stability. PMID- 15118041 TI - Knee stability following anterior cruciate ligament rupture and surgery. The contribution of irreducible tibial subluxation. AB - BACKGROUND: Knee stability after anterior cruciate ligament reconstruction is generally determined by measuring total anteroposterior tibial motion. In spite of a decrease in excessive anteroposterior tibial motion after anterior cruciate ligament reconstruction, problems can still develop. In the present study, we sought to define the tibiofemoral relationship more accurately with use of stress radiographs of human knees after anterior cruciate ligament rupture and after anterior cruciate ligament reconstruction. METHODS: A previously described radiographic technique was used to evaluate the position of the tibia relative to the femur with the application of an anteriorly directed tibial force and subsequently with the application of a posteriorly directed tibial force. Tibial position and total tibial translation were calculated from these radiographs. In addition, KT-1000 measurements were obtained. Three groups of patients were studied: Group 1 included twenty-eight patients with an untreated anterior cruciate ligament rupture, Group 2 included nineteen patients who had undergone a clinically successful anterior cruciate ligament reconstruction, and Group 3 included twenty-five control subjects with normal knees. RESULTS: KT-1000 testing showed that the average side-to-side differences in Group 1 (5.8 mm) and Group 2 (2.7 mm) were significantly different from that in Group 3 (0.8 mm) (p < 0.01 and p < 0.05, respectively). Stress radiographs showed that the average total tibial translation in Group 1 (9.8 mm) was significantly different from those in Group 2 (5.6 mm) and Group 3 (4.3 mm) (p < 0.05 and p < 0.001, respectively). Within Group 1, knees with radiographic signs of osteoarthritis were more stable, with an average total tibial excursion of 6.8 mm. The improved stability of the reconstructed knees in Group 2 and the osteoarthritic knees in Group 1 was not entirely the result of decreased anterior tibial translation; it was, in part, due to an irreducible anterior subluxation of the tibia. A posteriorly directed stress in these knees did not reduce the tibia to the anatomic position relative to the femur; the osteoarthritic knees in Group 1 were 9.9 mm short of full reduction and the knees in Group 2 were 3.1 mm short of full reduction (p < 0.01) CONCLUSIONS: Irreducible tibial subluxation can be present in the knee following surgical reconstruction of the anterior cruciate ligament. Osteoarthritic changes following an untreated anterior cruciate ligament rupture are also associated with uncorrectable tibial subluxation along with a decrease in instability. The irreducible tibial subluxation could explain why osteoarthritic changes still may develop in stable, reconstructed knees in spite of the improved stability. Currently used arthrometric measurements, such as KT-1000 scores, do not measure this phenomenon. PMID- 15118042 TI - Extracapsular placement of distal tibial transfixation wires. AB - BACKGROUND: Treatment of tibial plafond fractures with external fixation may involve use of transfixation wires within the periarticular region. Pin track infections that develop along wires placed intracapsularly may lead to joint infection. To our knowledge, there have been no previous investigations assessing the circumferential reflection of the ankle capsule or the potential for communication between the distal tibiofibular joint and the tibiotalar joint. The purpose of this study was to define these anatomic entities to provide guidelines for safe extracapsular placement of distal tibial wires. METHODS: Twelve fresh frozen cadaveric ankles and three ankles of living human volunteers were utilized for this study. High-resolution magnetic resonance imaging was performed on each ankle after pressurized distention of the joint capsule with gadolinium solution. The perpendicular distance from the subchondral bone at the joint line to the capsular synovial reflection was measured with use of a verified technique. The cadaveric ankles were sectioned, the capsular synovial reflections were measured by investigators who were blinded to the imaging results, and the corresponding measurements were compared. RESULTS: The anterolateral capsular synovial region displayed the most proximal reflection in all specimens (mean, 9.3 mm; maximum, 12.2 mm). The anteromedial region displayed less reflection (mean, 3.3 mm; maximum, 5.5 mm). All posteromedial and posterolateral synovial reflections were 12.2 mm from the subchondral surface of the plafond avoids penetration of the capsule. The distal tibiofibular joint communicates with the tibiotalar joint and thus should not be penetrated, to ensure extracapsular placement of the wires. PMID- 15118043 TI - The influence of the size and condition of the reamers on bone temperature during intramedullary reaming. AB - BACKGROUND: The objectives of this study were to determine whether intramedullary reaming increases bone temperature in vivo to a level that is high enough to produce bone necrosis and to determine the influence of the size and the condition of the reamers on the temperature increase. METHODS: Bone temperature variations were recorded in vivo during intramedullary tibial reaming in fourteen minipigs. The left tibiae were progressively reamed from 6 to 9 mm. The right tibiae were reamed with only 8 and 9-mm reamers. The variables studied were the initial and final temperature and the increase in the temperature. Two weeks after reaming, the tibiae were removed and studied histologically. RESULTS: Intramedullary reaming produced an average increase (and standard deviation) in bone temperature of 6.9 degrees C +/- 4.1 degrees C. The peak temperatures ranged from 34.9 degrees C to 49.4 degrees C. The average maximum bone temperature was 38.1 degrees C +/- 2.8 degrees C when the reaming was done progressively from 6 to 9 mm and 41.1 degrees C +/- 4.4 degrees C when the reaming was done only with the 8 and 9-mm reamers. The mean increase in the temperature in the second group of animals (8.2 degrees C +/- 4.3 degrees C) was greater than that in the first group (5.4 degrees C +/- 3.5 degrees C). Reaming with sharper reamers in the first seven animals resulted in a smaller mean increase in temperature than did reaming with less sharp reamers in the last seven animals (4.6 degrees C compared with 9.2 degrees C; p = 0.001). Histological examination of the tibiae showed periosteal proliferation and an altered disposition of the osteons at the inner cortex with occasional necrotic bone fragments in the medullary canal. CONCLUSIONS: Intramedullary reaming in the minipig increased bone temperature. When the reamer initially used was larger than the diameter of the medullary canal and when the reamers were blunted by repeated use, the maximum temperature reached by the bone was higher. This increase in temperature with use of typical medullary reaming techniques did not exceed the limits that would produce bone necrosis. PMID- 15118044 TI - A comparison of the microarchitectural bone adaptations of the concave and convex thoracic spinal facets in idiopathic scoliosis. AB - BACKGROUND: A limited number of studies have assessed the changes in bone microarchitecture in spinal facets with use of light microscopy but not with use of electron microscopy techniques. The purpose of this study was to analyze the facets in patients with scoliosis to determine whether there are differences in the bone microarchitecture of contralateral facets at the same anatomic level. METHODS: In eight patients undergoing posterior spinal arthrodesis for the treatment of idiopathic scoliosis, biopsy specimens of facet pairs at matched anatomic levels were obtained from three locations: (1). the curve apex, (2). one level cephalad to the apex, and (3). one level caudad to the apex. The facets were analyzed for cortical bone porosity and thickness with use of scanning electron microscopy and National Institutes of Health imaging software. The concave and convex facets were compared with use of a paired t test. RESULTS: The mean porosity (and standard deviation) for the concave and convex facets was 16.5% +/- 5.8% and 24.1% +/- 6.2%, respectively. Those on the convex side were significantly more porous than those on the concave side (p 1 month in the spinal cord of each animal. ES cell-derived axonal growth was inhibited by myelin, and this inhibition was overcome by administration of dibutyryl cAMP (dbcAMP) or a Rho kinase inhibitor in vitro and in vivo. In transplanted rats infused with dbcAMP, approximately 80 ES cell-derived motor axons were observed within the ventral roots of each animal, whereas none were observed in transplanted rats not treated with dbcAMP. Because these cells replicate many of the developmental and mature features of true motoneurons, they are an important biological tool to understand formation of motor units in vitro and a potential therapeutic tool to reconstitute neural circuits in vivo. PMID- 15118095 TI - Metabolic syndrome without obesity: Hepatic overexpression of 11beta hydroxysteroid dehydrogenase type 1 in transgenic mice. AB - In obese humans and rodents there is increased expression of the key glucocorticoid (GC) regenerating enzyme, 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1), in adipose tissue. This increased expression appears to be of pathogenic importance because transgenic mice overexpressing 11beta-HSD1 selectively in adipose tissue exhibit a full metabolic syndrome with visceral obesity, dyslipidemia, insulin-resistant diabetes, and hypertension. In this model, while systemic plasma GC levels are unaltered, GC delivery to the liver via the portal vein is increased. 11beta-HSD1 is most highly expressed in liver where inhibition or deficiency of its activity improves glucose and lipid homeostasis. To determine the potential contribution of elevated intrahepatic GCs alone toward development of insulin-resistant syndromes we generated transgenic mice expressing increased 11beta-HSD1 activity selectively in the liver under transcriptional control of hepatic regulatory sequences derived from the human apoE gene (apoE-HSD1). Transgenic lines with 2- and 5-fold-elevated 11beta-HSD1 activity exhibited mild insulin resistance without altered fat depot mass. ApoE HSD1 transgenic mice exhibited fatty liver and dyslipidemia with increased hepatic lipid synthesis/flux associated with elevated hepatic LXRalpha and PPARalpha mRNA levels as well as impaired hepatic lipid clearance. Further, apoE HSD1 transgenic mice have a marked, transgene-dose-associated hypertension paralleled by incrementally increased liver angiotensinogen expression. These data suggest that elevated hepatic expression of 11beta-HSD1 may relate to the pathogenesis of specific fatty liver, insulin-resistant, and hypertensive syndromes without obesity in humans as may occur in, for example, myotonic dystrophy, and possibly, the metabolically obese, normal-weight individual. PMID- 15118096 TI - Negative selection of semimature CD4(+)8(-)HSA+ thymocytes requires the BH3-only protein Bim but is independent of death receptor signaling. AB - T cell receptor/CD3 ligation induces apoptosis in semimature CD4(+)8(-)HSA+ thymocytes, and this helps establish immunological tolerance and constitutes one of the safeguards against autoimmune disease. We analyzed several knockout and transgenic mouse lines and found that T cell receptor/CD3-ligation-induced killing of semimature thymocytes occurred independently of Fas and "death receptor" signaling in general but required the proapoptotic BH3-only protein Bim and could be inhibited by Bcl-2. Loss of Apaf-1 or caspase-9, which act downstream of the Bcl-2 family protein family, provided only minor protection, indicating that the "apoptosome" functions as an amplifier rather than as an essential initiator of this death program. These results reveal the mechanisms of apoptosis in negative selection of semimature thymocytes and have implications for immunological tolerance and autoimmunity. PMID- 15118097 TI - Membrane protease proteomics: Isotope-coded affinity tag MS identification of undescribed MT1-matrix metalloproteinase substrates. AB - By proteolytic modification of low abundant signaling proteins and membrane receptors, proteases exert potent posttranslational control over cell behavior at the postsecretion level. Hence, substrate discovery is indispensable for understanding the biological role of proteases in vivo. Indeed, matrix metalloproteinases (MMPs), long associated with extracellular matrix degradation, are increasingly recognized as important processing enzymes of bioactive molecules. MS is now the primary proteomic technique for detecting, identifying, and quantitating proteins in cells or tissues. Here we used isotopecoded affinity tag labeling and multidimensional liquid chromatography inline with tandem MS to identify MDA-MB-231 breast carcinoma cell proteins shed from the cell surface or the pericellular matrix and extracellular proteins that were degraded or processed after transfection with human membrane type 1-MMP (MT1-MMP). Potential substrates were identified as those having altered protein levels compared with the E240A inactive MT1-MMP mutant or vector transfectants. New substrates were biochemically confirmed by matrix-assisted laser desorption ionization-time-of flight MS and Edman sequencing of cleavage fragments after incubation with recombinant soluble MT1-MMP in vitro. We report many previously uncharacterized substrates of MT1-MMP, including the neutrophil chemokine IL-8, secretory leukocyte protease inhibitor, pro-tumor necrosis factor alpha, death receptor-6, and connective tissue growth factor, indicating that MT1-MMP is an important signaling protease in addition to its traditionally ascribed roles in pericellular matrix remodeling. Moreover, the high-throughput and quantitative nature of isotope-coded affinity tag labeling combined with tandem MS sequencing is a previously undescribed degradomic screen for protease substrate discovery that should be generally adaptable to other classes of protease for exploring proteolytic function in complex and dynamic biological contexts. PMID- 15118098 TI - Modular construction of a signaling scaffold: MORG1 interacts with components of the ERK cascade and links ERK signaling to specific agonists. AB - Signal transduction occurs by the reversible assembly of oligomeric protein complexes that include both enzymatic proteins and proteins without known enzymatic activity. These nonenzymatic components can serve as scaffolds or anchors and regulate the efficiency, specificity, and localization of the signaling pathway. Here we report the identification of MORG1 (mitogen-activated protein kinase organizer 1), a member of the WD-40 protein family that was isolated as a binding partner of the extracellular signal-regulated kinase (ERK) pathway scaffold protein MP1. MORG1 specifically associates with several components of the ERK pathway, including MP1, Raf-1, MEK, and ERK, and stabilizes their assembly into an oligomeric complex. MORG1 facilitates ERK activation when cells are stimulated with lysophosphatidic acid, phorbol 12-myristate 13-acetate, or serum, but not in response to epidermal growth factor. Suppression of MORG1 by short interfering RNA leads to a marked reduction in ERK activity when cells are stimulated with serum. We propose that MORG1 is a component of a modular scaffold system that participates in the regulation of agonist-specific ERK signaling. PMID- 15118099 TI - Development of innate CD4+ alpha-chain variable gene segment 24 (Valpha24) natural killer T cells in the early human fetal thymus is regulated by IL-7. AB - Natural killer (NK) T cells are innate CD1d-restricted immune cells involved in regulation of immune tolerance, tumor immunity, and immunity to infectious pathogens. Human alpha-chain variable gene segment 24 (Valpha24) NK T cells exist in the periphery as two functionally distinct subsets: one CD4+ and one CD4- subset. However, the developmental pathway of human Valpha24 NK T cells is not well understood. Here, we show that Valpha24 NK T cells develop in the fetal thymus. The relative number of intrathymic NK T cell precursors decline in a linear manner with gestational age, and they are very rare in the neonatal thymus, indicating that these cells preferentially develop in the early fetal thymus. Their restriction element, CD1d, is expressed by a vast majority of thymocytes. A majority of intrathymic Valpha24 NK T cell progenitors are CD4+, whereas a minority are CD4/8(+/+). CD4+ Valpha24 NK T cell precursors show features of mature NK T cells, such as high levels of their semiinvariant T cell receptor and CD3 and some expression of CD161, whereas the CD4/8(+/+) precursors seem less mature. The cytokine IL-7 shows a biphasic effect on Valpha24 NK T cell progenitors in fetal thymic organ culture, with high doses driving proliferation of immature CD161-progenitors and low doses supporting survival and maturation. Thus, the data demonstrate that human Valpha24 NK T cells of the CD4+, but not the CD4-, subset develop in the early fetal thymus. Furthermore, data suggest an intrathymic pathway of CD4+ Valpha24 NK T cell development that is regulated by IL-7. PMID- 15118100 TI - Structural and functional analysis of Aplysia attractins, a family of water-borne protein pheromones with interspecific attractiveness. AB - Mate attraction in Aplysia involves a long-distance water-borne signal (the protein pheromone attractin), which is released during egg laying. Aplysia californica attractin attracts species that produce closely related attractins, such as Aplysia brasiliana, whose geographic distribution does not overlap that of A. californica. This finding suggests that other mollusks release attractin related pheromones to form and maintain breeding aggregations. We describe four additional members of the attractin family: A. brasiliana, Aplysia fasciata, Aplysia depilans (which aggregates with A. fasciata aggregations), and Aplysia vaccaria (which aggregates with A. californica aggregations). On the basis of their sequence similarity with A. californica attractin, the attractin proteins fall into two groups: A. californica, A. brasiliana, and A. fasciata (91-95% identity), and A. depilans and A. vaccaria (41-43% identity). The sequence similarity within the attractin family, the conserved six cysteines, and the compact fold of the NMR solution structure of A. californica attractin suggest a common fold for this pheromone family containing two antiparallel helices. The second helix contains the IEECKTS sequence conserved in Aplysia attractins. Mutating surface-exposed charged residues within this heptapeptide sequence abolishes attractin activity, suggesting that the second helix is an essential part of the receptor-binding interface. PMID- 15118101 TI - Sensory neuron-specific receptor activation elicits central and peripheral nociceptive effects in rats. AB - The sensory neuron-specific G protein coupled receptors (SNSRs) have been described as a family of receptors whose expression in small diameter sensory neurons in the trigeminal and dorsal root ganglia suggests an implication in nociception. To date, the physiological function(s) of SNSRs remain unknown. Hence, the aim of the present study was to determine the effects of rat SNSR1 activation on nociception in rats. The pharmacological characterization of rat SNSR1 was initially performed in vitro to identify a specific ligand, which could be used subsequently in the rat for physiological testing. Among all ligands tested, gamma2-MSH was the most potent at activating rat SNSR1. Structure activity relationship studies revealed that the active moiety recognized by rat SNSR1 was the C-terminal part of gamma2-MSH. The radiolabeled C-terminal part of gamma2-MSH, gamma2-MSH-6-12, bound with high affinity to membranes derived from rat skin and spinal cord, demonstrating the presence of receptor protein at both the proximal and distal terminals of dorsal root ganglia. To investigate the physiological role of SNSR, specific ligands to rat SNSR1 were tested in behavioral assays of pain sensitivity in rats. Selective rat SNSR1 agonists produced spontaneous pain behavior, enhanced heat and mechanical sensitivity when injected intradermally, and heat hypersensitivity when injected centrally, consistent with the localization of rat SNSR1 protein at central and peripheral sites. Together, these results clearly indicate that the SNSR1 plays a role in nociception and may provide novel therapeutic opportunities for analgesia. PMID- 15118102 TI - Mechanism of homophilic adhesion by the neural cell adhesion molecule: use of multiple domains and flexibility. AB - The extracellular regions of adhesion proteins of the Ig superfamily comprise multiple, tandemly arranged domains. We used directforce measurements to investigate how this modular architecture contributes to the adhesive interactions of the neural cell adhesion molecule (NCAM), a representative of this protein class. The extracellular region of NCAM comprises five immunoglobulin and two fibronectin domains. Previous investigations generated different models for the mechanism of homophilic adhesion that each use different domains. We use force measurements to demonstrate that NCAM binds in two spatially distinct configurations. Igdomain deletion mutants identified the domains responsible for each of the adhesive bonds. The measurements also confirmed the existence of a flexible hinge that alters the orientation of the adhesive complexes and the intermembrane distance. These results suggest that a combination of multiple bound states and internal molecular flexibility allows for sequentially synergistic bond formation and the ability to accommodate differences in intercellular space. PMID- 15118103 TI - Close agreement between the orientation dependence of hydrogen bonds observed in protein structures and quantum mechanical calculations. AB - Hydrogen bonding is a key contributor to the exquisite specificity of the interactions within and between biological macromolecules, and hence accurate modeling of such interactions requires an accurate description of hydrogen bonding energetics. Here we investigate the orientation and distance dependence of hydrogen bonding energetics by combining two quite disparate but complementary approaches: quantum mechanical electronic structure calculations and protein structural analysis. We find a remarkable agreement between the energy landscapes obtained from the electronic structure calculations and the distributions of hydrogen bond geometries observed in protein structures. In contrast, molecular mechanics force fields commonly used for biomolecular simulations do not consistently exhibit close correspondence to either quantum mechanical calculations or experimentally observed hydrogen bonding geometries. These results suggest a route to improved energy functions for biological macromolecules that combines the generality of quantum mechanical electronic structure calculations with the accurate context dependence implicit in protein structural analysis. PMID- 15118104 TI - Dual architectural roles of HU: formation of flexible hinges and rigid filaments. AB - The nucleoid-associated protein HU is one of the most abundant proteins in Escherichia coli and has been suggested to play an important role in bacterial nucleoid organization and regulation. Although the regulatory aspects of HU have been firmly established, much less is understood about the role of HU in shaping the bacterial nucleoid. In both functions (local) modulation of DNA architecture seems an essential feature, but information on the mechanical properties of this type of sequence-independent nucleoprotein complex is scarce. In this study we used magnetic tweezers and atomic force microscopy to quantify HU-induced DNA bending and condensation. Both techniques revealed that HU can have two opposing mechanical effects depending on the protein concentration. At concentrations <100 nM, individual HU dimers induce very flexible bends in DNA that are responsible for DNA compaction up to 50%. At higher HU concentrations, a rigid nucleoprotein filament is formed in which HU appears to arrange helically around the DNA without inducing significant condensation. PMID- 15118105 TI - Imaging correlates of brain function in monkeys and rats isolates a hippocampal subregion differentially vulnerable to aging. AB - The hippocampal formation contains a distinct population of neurons organized into separate anatomical subregions. Each hippocampal subregion expresses a unique molecular profile accounting for their differential vulnerability to mechanisms of memory dysfunction. Nevertheless, it remains unclear which hippocampal subregion is most sensitive to the effects of advancing age. Here we investigate this question by using separate imaging techniques, each assessing different correlates of neuronal function. First, we used MRI to map cerebral blood volume, an established correlate of basal metabolism, in the hippocampal subregions of young and old rhesus monkeys. Second, we used in situ hybridization to map Arc expression in the hippocampal subregions of young and old rats. Arc is an immediate early gene that is activated in a behavior-dependent manner and is correlated with spike activity. Results show that the dentate gyrus is the hippocampal subregion most sensitive to the effects of advancing age, which together with prior studies establishes a cross-species consensus. This pattern isolates the locus of age-related hippocampal dysfunction and differentiates normal aging from Alzheimer's disease. PMID- 15118106 TI - Genome-wide transcription profile of field- and laboratory-selected dichlorodiphenyltrichloroethane (DDT)-resistant Drosophila. AB - Genome-wide microarray analysis (Affymetrix array) was used (i) to determine whether only one gene, the cytochrome P450 enzyme Cyp6g1, is differentially transcribed in dichlorodiphenyltrichloroethane (DDT)-resistant vs. -susceptible Drosophila; and (ii) to profile common genes differentially transcribed across a DDT-resistant field isolate [Rst(2)DDT(Wisconsin)] and a laboratory DDT-selected population [Rst(2)DDT(91-R)]. Statistical analysis (ANOVA model) identified 158 probe sets that were differentially transcribed among Rst(2)DDT(91-R), Rst(2)DDT(Wisconsin), and the DDT-susceptible genotype Canton-S (P < 0.01). The cytochrome P450 Cyp6a2 and the diazepam-binding inhibitor gene (Dbi) were over transcribed in the two DDT-resistant genotypes when compared to the wild-type Drosophila, and this difference was significant at the most stringent statistical level, a Bonferroni correction. The list of potential candidates differentially transcribed also includes 63 probe sets for which molecular function ontology annotation of the probe sets did not exist. A total of four genes (Cyp6a2, Dbi, Uhg1, and CG11176) were significantly different (P < 5.6 e(-06)) between Rst(2)DDT(91-R) and Canton-S. Additionally, two probe sets encoding Cyp12d1 and Dbi were significantly different between Rst(2)DDT(Wisconsin) and Canton-S after a Bonferroni correction. Fifty-two probe sets, including those associated with pesticide detoxification, ion transport, signal transduction, RNA transcription, and lipid metabolism, were commonly expressed in both resistant lines but were differentially transcribed in Canton-S. Our results suggest that more than Cyp6g1 is overtranscribed in field and laboratory DDT-resistant genotypes, and the number of commonalities suggests that similar resistance mechanisms may exist between laboratory- and field-selected DDT-resistant fly lines. PMID- 15118107 TI - Identification of an inhibitor-binding site to HIV-1 integrase with affinity acetylation and mass spectrometry. AB - We report a methodology that combines affinity acetylation with MS analysis for accurate mapping of an inhibitor-binding site to a target protein. For this purpose, we used a known HIV-1 integrase inhibitor containing aryl di-O-acetyl groups (Acetylated-Inhibitor). In addition, we designed a control compound (Acetylated-Control) that also contained an aryl di-O-acetyl group but did not inhibit HIV-1 integrase. Examination of the reactivity of these compounds with a model peptide library, which collectively contained all 20 natural amino acids, revealed that aryl di-O-acetyl compounds effectively acetylate Cys, Lys, and Tyr residues. Acetylated-Inhibitor and Acetylated-Control exhibited comparable chemical reactivity with respect to these small peptides. However, these two compounds differed markedly in their interactions with HIV-1 integrase. In particular, Acetylated-Inhibitor specifically acetylated K173 at its inhibitory concentration (3 microM) whereas this site remained unrecognized by Acetylated Control. Our data enabled creation of a detailed model for the integrase:Acetylated-Inhibitor complex, which indicated that the inhibitor selectively binds at an architecturally critical region of the protein. The methodology reported herein has a generic application for systems involving a variety of ligand-protein interactions. PMID- 15118108 TI - PIKE-A is amplified in human cancers and prevents apoptosis by up-regulating Akt. AB - PIKE-A (PIKE-activating Akt), an isoform of PIKE GTPase that enhances phosphatidylinositol 3-kinase (PI3-kinase) activity, specifically binds to active Akt but not PI3-kinase. PIKE-A stimulates Akt activity in a GTP-dependent manner and promotes invasiveness of cancer cell lines. Here, we show that PIKE-A is amplified in a variety of human cancers and that amplified PIKE-A directly stimulates Akt and inhibits apoptosis compared to cells with normal PIKE-A copy number. Overexpression of PIKE-A wild-type but not dominant-negative mutant stimulates Akt activity and prevents apoptosis. Moreover, knockdown of PIKE-A diminishes Akt activity and increases apoptosis. Our findings suggest that PIKE-A amplification contributes to cancer cell survival and progression by inhibiting apoptosis through up-regulating Akt. PMID- 15118109 TI - The cleft lip and palate defects in Dancer mutant mice result from gain of function of the Tbx10 gene. AB - Cleft lip and palate (CL/P) is a common disfiguring birth defect with complex, poorly understood etiology. Mice carrying a spontaneous mutation, Dancer (Dc), exhibit CL/P in homozygotes and show significantly increased susceptibility to CL/P in heterozygotes [Deol, M. S. & Lane, P. W. (1966) J. Embryol. Exp. Morphol. 16, 543-558 and Trasler, D. G., Kemp, D. & Trasler, T. A. (1984) Teratology 29, 101-104], providing an animal model for understanding the molecular pathogenesis of CL/P. We genetically mapped Dc to within a 1-cM region near the centromere of chromosome 19. In situ hybridization analysis showed that one positional candidate gene, Tbx10, is ectopically expressed in Dc mutant embryos. Positional cloning of the Dc locus revealed an insertion of a 3.3-kb sequence containing the 5' region of the p23 gene into the first intron of Tbx10, which causes ectopic expression of a p23-Tbx10 chimeric transcript encoding a protein product identical to a normal variant of the Tbx10 protein. Furthermore, we show that ectopic expression of Tbx10 in transgenic mice recapitulates the Dc mutant phenotype, indicating that CL/Pin Dc mutant mice results from the p23 insertion induced ectopic Tbx10 expression. These results identify gain of function of a T box transcription factor gene as a mechanism underlying CL/P pathogenesis. PMID- 15118111 TI - Bladder cancer: analysis of multi-detector row helical CT enhancement pattern and accuracy in tumor detection and perivesical staging. AB - PURPOSE: To evaluate the enhancement pattern of bladder cancer and the accuracy of multi-detector row helical computed tomography (CT) in the detection and staging of bladder cancer. MATERIALS AND METHODS: In 20 patients, the attenuation value of bladder cancer was measured on dynamic contrast material-enhanced multiphasic CT images obtained with scanning delays of 40, 60, 80, and 100 seconds. In 67 patients, CT data were obtained with a 60-second scanning delay that covered the bladder (section thickness, 2.5 mm; beam pitch, 1.5) and a 180 second scanning delay that covered the abdomen (section thickness, 5 mm; beam pitch, 1.5). We prospectively evaluated CT images and compared findings at CT with findings at histologic examination. We evaluated cancer detection rate, positive predictive value of cancer detection, and sensitivity and specificity in the diagnosis of perivesical invasion. RESULTS: The attenuation value of bladder cancers was significantly higher on 60- (105 HU +/- 16) and 80-second (97 HU +/- 15) delayed CT images than on the other images (P <.05). The cancer detection rate and positive predictive value for cancer detection were 97% and 95%, respectively, in 67 patients and increased to 100% and 100%, respectively, in 44 patients with a time interval of 7 or more days between transurethral resection of the bladder (TURB) and CT examination. Sensitivity and specificity in the diagnosis of perivesical invasion were 89% and 95%, respectively, in 67 patients and increased to 92% and 98%, respectively, in 44 patients with a time interval of 7 or more days between TURB and CT examination. CONCLUSION: Bladder cancer tends to show peak enhancement with the 60-second scanning delay. Multi-detector row helical CT is useful in the detection and staging of bladder cancer. PMID- 15118110 TI - Cerebral perfusion CT: technique and clinical applications. AB - Perfusion computed tomography (CT) is a relatively new technique that allows rapid qualitative and quantitative evaluation of cerebral perfusion by generating maps of cerebral blood flow (CBF), cerebral blood volume (CBV), and mean transit time (MTT). The technique is based on the central volume principle (CBF = CBV/MTT) and requires the use of commercially available software employing complex deconvolution algorithms to produce the perfusion maps. Some controversies exist regarding this technique, including which artery to use as input vessel, the accuracy of quantitative results, and the reproducibility of results. Despite these controversies, perfusion CT has been found to be useful for noninvasive diagnosis of cerebral ischemia and infarction and for evaluation of vasospasm after subarachnoid hemorrhage. Perfusion CT has also been used for assessment of cerebrovascular reserve by using acetazolamide challenge in patients with intracranial vascular stenoses who are potential candidates for bypass surgery or neuroendovascular treatment, for the evaluation of patients undergoing temporary balloon occlusion to assess collateral flow and cerebrovascular reserve, and for the assessment of microvascular permeability in patients with intracranial neoplasms. This article is a review of the technique, clinical applications, and controversies surrounding perfusion CT. PMID- 15118112 TI - Nontuberculous mycobacterial pulmonary infection in immunocompetent patients: comparison of thin-section CT and histopathologic findings. AB - PURPOSE: To identify and describe the thin-section computed tomographic (CT) findings of nontuberculous mycobacterial (NTM) pulmonary infection in immunocompetent patients and to compare these findings with histopathologic findings. MATERIALS AND METHODS: Between April 2002 and March 2003, the thin section chest CT findings in and histopathologic lung tissue specimens from 22 patients who fulfilled the American Thoracic Society diagnostic criteria for NTM pulmonary infection were retrospectively reviewed. The lung lesion patterns (ie, small nodules, branching centrilobular nodules [ie, tree-in-bud pattern], consolidation, cavities, bronchiectasis, and volume loss) seen at CT at the sites of transbronchial lung biopsy (n = 22) or lobectomy (n = 1) were compared with the histopathologic findings. RESULTS: Thirteen of the 22 patients were found to have Mycobacterium abscessus pulmonary infection; seven, to have Mycobacterium avium-intracellulare complex infection; and two, to have Mycobacterium fortuitum infection. Regardless of the specific infective mycobacterial species, bilateral small nodules (in 22 [100%] lung locations), cylindric bronchiectasis (in 20 [91%] locations), and branching centrilobular nodular lesions (in 17 [77%] locations) were the most common CT findings seen at the biopsy sites. All of the transbronchial lung biopsy specimens showed a thickened bronchiolar wall and bronchiolar and peribronchiolar inflammation at histopathologic analysis. Dilated bronchioles were identified in 19 (86%) patients, and epithelioid granulomas with or without caseation were seen in seven (32%). CONCLUSION: Regardless of the specific infective mycobacterial species, the most common thin-section CT findings of NTM pulmonary infection are bilateral small nodules, cylindric bronchiectasis, and branching centrilobular nodules. These findings correspond histopathologically to bronchiolectasis and bronchiolar and peribronchiolar inflammation with or without granuloma formation. PMID- 15118113 TI - Chemical shift MR imaging of hyperattenuating (>10 HU) adrenal masses: does it still have a role? AB - PURPOSE: To evaluate chemical shift magnetic resonance (MR) imaging for the characterization of hyperattenuating adrenal masses. MATERIALS AND METHODS: Adrenal MR images obtained from January 1998 to February 2003 were reviewed. Patients were excluded if they did not undergo unenhanced computed tomography or did not have an adrenal mass with attenuation higher than 10 HU, adequate follow up, or pathologic diagnosis for use as a reference standard. A diagnosis of adenoma required at least 24 weeks of stability on images. Thirty-eight masses in 36 patients were identified (27 adenomas, nine metastases, one adrenocortical oncocytoma, and one pheochromocytoma). Signal intensity (SI) decrease between in phase and opposed-phase MR images was measured for the entire mass and normalized to the renal parenchymal SI. In 21 of 36 (58%) patients, dual-echo single-breath hold MR imaging was used to eliminate misregistration. RESULTS: The attenuation of 61% (23 of 38) of all masses and 70% (19 of 27) of adenomas was 10-30 HU. With a threshold of 20% SI decrease, the sensitivity of chemical shift MR imaging for hyperattenuating adenoma was 67% (18 of 27 masses). When considering masses with attenuation of 10-30 HU, the sensitivity for adenoma was 89% (17 of 19 masses) and remained reasonable at 75% (six of eight masses) for adenomas with attenuation of 20-30 HU. Only one adenoma with attenuation higher than 30 HU had SI decrease of more than 20%. Specificity for diagnosis of adenoma was 100% (11 of 11). CONCLUSION: In certain circumstances, chemical shift MR imaging is a reasonable second imaging test for further characterization of a hyperattenuating adrenal mass. PMID- 15118114 TI - Evidence for spleen-specific uptake of a microbubble contrast agent: a quantitative study in healthy volunteers. AB - PURPOSE: To evaluate the pharmacokinetics of the microbubble contrast agent BR1. MATERIALS AND METHODS: Twenty healthy volunteers were injected via arm vein with a 1.2-mL bolus of BR1. Ultrasonographic images of liver and right kidney and of spleen and left kidney were obtained intermittently for 5 minutes with low mechanical-index software (to minimize microbubble destruction) that shows stationary microbubbles in green. Percentage total uptake was calculated as the number of green pixels in the region of interest for each organ over time, divided by the total pixels. Relative uptake, the ratio of total uptake in liver to that in right kidney and of total uptake in spleen to that in left kidney, and differential uptake, the difference in total uptake between liver and right kidney and between spleen and left kidney, were calculated. Total uptake for each organ was plotted against time, and the gradient of a best-fit straight line was calculated. Wilcoxon signed rank test was used to compare mean uptake values in each subject. Mann-Whitney U test was used for comparisons in sex and age. RESULTS: Total uptake declined over 5 minutes in left and right kidney and in liver (from 88% +/- 10% [1 minute] to 67% +/- 14% [5 minutes]), but not in spleen (range, 90%-99%). Mean relative uptake +/- 1 SD for spleen increased from 2.3 +/- 0.7 (1 minute) to 3.7 +/- 2.3 (5 minutes) (P =.005) but for liver was constant: 2.1 +/- 0.9 (1 minute) and 2.3 +/- 0.4 (5 minutes) (P =.06). Mean differential uptake +/- 1 SD for spleen increased from 51.3% +/- 14.9% (1 minute) to 65.0% +/- 9.1% (5 minutes) (P =.002). Significant difference was seen over time in total uptake gradients between spleen and left kidney (P =.014) but not between liver and right kidney or right and left kidney. No difference was seen between men and women or with age. CONCLUSION: BR1 produces spleen-specific enhancement that is longer (5 minutes) than the blood pool phase. PMID- 15118115 TI - Relation between aneurysm volume, packing, and compaction in 145 cerebral aneurysms treated with coils. AB - PURPOSE: To assess the relation between aneurysm volume, packing, and compaction in cerebral aneurysms treated with coils. MATERIALS AND METHODS: The volumes of 145 aneurysms that were treated with coils were calculated with biplanar angiographic images and a custom-designed method. Partially thrombosed aneurysms were excluded. Packing was defined as the ratio between the volume of the inserted coils and the volume of the aneurysm and was calculated for all 145 aneurysms. Results at 6-month follow-up angiography were dichotomized into presence or absence of compaction. RESULTS: Aneurysm volume, packing, and compaction at 6-month follow-up were closely related. Large aneurysm volume was associated with low packing and frequent compaction. High packing prevents compaction. If the aneurysm volume was packed for 24% or more with coils, compaction did not occur in aneurysms with a volume of less than 600 mm(3). In small aneurysms with volumes of less than 200 mm(3), compaction did not occur when packing was above 20%. CONCLUSION: The common practice of inserting as many coils as possible in cerebral aneurysms is sensible in trying to avoid compaction. In aneurysms with packing of 24% or more, no compaction occurred at 6 month angiographic follow-up. In aneurysms with a volume of more than 600 mm(3), high packing could not be achieved, which resulted in compaction in the majority of aneurysms. PMID- 15118116 TI - Molecular targeting of lymph nodes with L-selectin ligand-specific US contrast agent: a feasibility study in mice and dogs. AB - PURPOSE: To evaluate the feasibility of using intravenously administered L selectin ligand-specific polymer-stabilized air-filled microparticles (MPs) for active targeting of peripheral lymph nodes under normal conditions in animal models. MATERIALS AND METHODS: L-selectin ligand-specific MPs and two control substances (immunoglobulin M-isotype MPs and native MPs) were each administered in three conscious mice as a single intravenous bolus injection (1.4 x 10(7) MPs/kg). All mice were sacrificed 30 minutes after administration. Lymph nodes (cervical, inguinal, axillary, popliteal, mesenteric), spleen (positive control), and kidney (blood pool control) were removed and examined for MP-related stimulated acoustic emission (SAE) signals by using harmonic color Doppler ultrasonography (US) in a tank containing degassed water. A second experiment was performed in six anesthetized beagle dogs by using the same MP formulation. Each of the MP formulations was administered in two anesthetized dogs as a single intravenous bolus injection (1 x 10(7) MPs/kg). The popliteal lymph nodes, spleen (positive control), and kidney (blood pool control) were examined in vivo with US for MP-related SAE signals 30 minutes after administration. Fisher exact test for the one-side alternative was used for mouse data analysis. RESULTS: The lymph nodes of all mice (P =.05) and the popliteal lymph nodes of both dogs treated with L-selectin ligand-specific MPs showed clear MP-related SAE signals, whereas the lymph nodes of all mice and the popliteal lymph nodes of four dogs that received the control substances did not show any SAE signals. CONCLUSION: Use of an intravenously administered L-selectin ligand-specific US contrast agent is feasible for active lymph node targeting in mice and dogs. PMID- 15118117 TI - Polytetrafluoroethylene-covered nitinol stent-graft for transjugular intrahepatic portosystemic shunt creation: 3-year experience. AB - PURPOSE: To prospectively evaluate the use of a recently developed expanded polytetrafluoroethylene (PTFE)-covered nitinol stent-graft in preventing the need for repeated intervention after transjugular intrahepatic portosystemic shunt (TIPS) creation. MATERIALS AND METHODS: Fifty-three consecutive patients underwent TIPS procedures between January 2000 and February 2002. Minimum patient follow-up was 9 months (mean, 16.3 months). Fifty-six stent-grafts were implanted in 53 patients; eight of the devices were 8 mm in diameter and 48 were 10 mm in diameter. The stent length varied from 4 to 7 cm. Indications for the procedure included recurrence of bleeding after sclerotherapy (28 patients with cirrhosis, one patient without), refractory ascites or hydrothorax (21 patients with cirrhosis, one patient without), and Budd-Chiari syndrome (two patients). RESULTS: A technical success rate of 100% was obtained, with an early clinical success rate of 96.2%. During the follow-up period, the recurrence rate was 3.4% (one of 29 patients) for bleeding and 9.0% (two of 22 patients) for ascites. Shunt malfunction occurred in nine of 53 patients (16.9%); in one of these nine patients, shunt occlusion was evident after revision, and a parallel shunt was created. The 1-year primary and secondary patency rates were 83.8% and 98.1%, respectively. In this series, the incidence of encephalopathy (included even as a single short-lived episode) was 47.1% (25 of 53 patients). The 30-day mortality rate was 3.8% (two of 53), and the late mortality rate was 17.3% (eight of 46), excluding seven patients who underwent transplantation. CONCLUSION: The new PTFE covered nitinol stent-graft used appears to be excellent in preventing the need for repeated interventions. A primary patency rate of 83.8% and a secondary patency rate of 98.1% were achieved. PMID- 15118118 TI - Acute myocardial infarction: contrast-enhanced multi-detector row CT in a porcine model. AB - PURPOSE: To assess the role of contrast material-enhanced retrospectively electrocardiographically (ECG) gated multi-detector row computed tomography (CT) in the detection of acute myocardial infarction in a porcine model of total coronary occlusion. MATERIALS AND METHODS: Seven Yorkshire farm pigs were studied with contrast-enhanced retrospectively ECG-gated multi-detector row CT 3 hours after total occlusion of the distal left anterior descending artery (n = 5) or the second diagonal branch (n = 2). Reformatted short-axis end-systolic and end diastolic CT data sets were assessed for myocardial perfusion deficits, coronary occlusion, and abnormal myocardial wall motion. Perfusion deficits were compared with microsphere-determined blood flow and triphenyltetrazolium chloride (TTC) stained tissue samples for infarct assessment by using Bland-Altman analysis and analysis of variance. RESULTS: Myocardial perfusion deficits, occlusion of the left anterior descending artery or second diagonal branch, and akinesis of the infarcted segment were identified in all five animals that completed the study. One animal died, and one data set had nondiagnostic image quality. The CT end diastolic (mean, 16.1% +/- 4.8 [SD]; range, 8.6%-22.2%) and end-systolic (mean, 17.0% +/- 6.4; range, 8.7%-26.8%) volume of perfusion deficit was similar to that of infarcted tissue at TTC staining (mean, 13.6% +/- 6.0; range, 7.8%-30.9%). Infarcted myocardium at CT demonstrated a 76.1% reduction in microsphere determined blood flow and a significant reduction of myocardial CT attenuation compared with normal myocardium (P <.01). Myocardial wall motion analysis demonstrated absence of systolic wall thickening in infarcted myocardium. CONCLUSION: Multi-detector row CT with retrospective ECG gating permits the detection and further characterization of acute myocardial infarction in a porcine model of complete coronary occlusion. PMID- 15118119 TI - Carotid perfusion CT with balloon occlusion and acetazolamide challenge test: feasibility. AB - Carotid balloon test occlusion (BTO) is used to assess the collateral circulation and cerebrovascular reserve in patients in whom carotid artery occlusion is contemplated. Eight patients in whom the test was successful were evaluated with perfusion computed tomography (CT) in the resting state and after acetazolamide challenge. Three of the patients showed symmetric blood flow and normal response to acetazolamide. One of them underwent permanent carotid occlusion and did not develop any delayed ischemic stroke. The remaining five patients showed asymmetric blood flow. One of them had markedly low blood flow and abnormal response to acetazolamide. The patient developed ipsilateral hemispheric stroke following permanent carotid occlusion after the superficial temporal artery to middle cerebral artery bypass graft occluded. In the other four patients, the steal phenomenon was seen in ipsilateral and contralateral hemispheres. Although definitive quantitative values for perfusion CT are not yet standardized, it may be feasible to predict that the patients with symmetric blood flow and normal acetazolamide-enhanced challenge test results will do well after permanent carotid occlusion. Patients with asymmetric blood flow and abnormal response to the acetazolamide challenge test may require a revascularization procedure to protect them from delayed ischemic stroke. PMID- 15118120 TI - Quantitative and qualitative evaluations of fetal lung with MR imaging. AB - PURPOSE: To measure both volume and signal intensity of the fetal lung at magnetic resonance (MR) imaging and to evaluate the clinical use of this method to predict fetal pulmonary hypoplasia. MATERIALS AND METHODS: A total of 87 fetuses evaluated with MR imaging at 24-39 weeks of gestation were classified into a control group with good respiratory outcome (group A, n = 58) or a poor outcome group with severe respiratory disturbance after birth (group B, n = 29). Planimetric measurement of total lung volume and calculation of the ratio of lung signal intensity to spinal fluid signal intensity (L/SF) were performed on MR images by using region-of-interest analysis. Regression analysis, analysis of covariance, analysis of variance, and receiver operating characteristic (ROC) analysis were performed. RESULTS: The best fit for group A lung volume was represented by the regression line V = (2.41 x G) - 37.6 (r = 0.537, P <.001), in which V is lung volume and G is gestational weeks; that for group B, by V = (0.97 x G) - 14.0 (r = 0.378, P <.05). Results of analysis of covariance with gestational weeks used as a covariate showed a significant difference in lung volume between the two groups (P <.001). Mean +/- SEM for L/SF ratio was 0.817 +/ 0.013 and 0.598 +/- 0.019 in groups A and B, respectively (P <.001). For prediction of postnatal respiratory outcome, the area under the ROC curve for lung volume and L/SF ratio combined was 0.990, significantly higher than that for lung volume alone (P <.05). CONCLUSION: Simultaneous measurement of fetal lung volume and signal intensity on MR images is a promising method for predicting fetal pulmonary hypoplasia. PMID- 15118121 TI - Infrared spectroscopic evidence for protonated water clusters forming nanoscale cages. AB - Size-dependent development of the hydrogen bond network structure in large sized clusters of protonated water, H+(H2O)n (n = 4 to 27), was probed by infrared spectroscopy of OH stretches. Spectral changes with cluster size demonstrate that the chain structures at small sizes (n less, similar 10) develop into two dimensional net structures (approximately 10 < n < 21), and then into nanometer scaled cages (n >/= 21). PMID- 15118122 TI - Infrared signature of structures associated with the H+(H2O)n (n = 6 to 27) clusters. AB - We report the OH stretching vibrational spectra of size-selected H+(H2O)n clusters through the region of the pronounced "magic number" at n = 21 in the cluster distribution. Sharp features are observed in the spectra and assigned to excitation of the dangling OH groups throughout the size range 6 or =140% or 12.5 cm3. RESULTS: Hematoma enlargement occurred in 16 patients. Maximum SBP was significantly associated with hematoma enlargement (P=0.0074). A logistic regression model for predicting hematoma enlargement was constructed with the use of maximum SBP, hematoma volume, and Glasgow Coma Scale score at admission. After adjustment for these factors, maximum SBP was independently associated with hematoma enlargement (odds ratio per mm Hg, 1.04; 95% CI, 1.01 to 1.07). Target SBPs of > or =160 mm Hg were significantly associated with hematoma enlargement compared with those of < or =150 mm Hg (P=0.025). CONCLUSIONS: Our findings suggest that elevated BP increases the risk of hematoma enlargement. Efforts to lower SBP below 150 mm Hg may prevent this risk. PMID- 15118170 TI - Cigarette smoking and risk of stroke and its subtypes among middle-aged Japanese men and women: the JPHC Study Cohort I. AB - BACKGROUND AND PURPOSE: We examined sex-specific relationships of smoking with risk of total stroke and stroke subtypes in Asian populations because of the limited data available. METHODS: A total of 19,782 men and 21,500 women aged 40 to 59 years who were free of prior diagnosis of stroke, coronary heart disease, or cancer and reported their smoking status were followed in the Japan Public Health Center-based Prospective Study on Cancer and Cardiovascular Disease (JPHC Study) from 1990 to 1992 to the end of 2001. RESULTS: During a 461,761 person year follow-up, 702 total strokes were documented among men, of which 619 were confirmed by imaging studies, including 219 intraparenchymal hemorrhages, 73 subarachnoid hemorrhages, and 327 ischemic strokes. The respective numbers of cases among women were 447, 411, 129, 106, and 176. Multivariate relative risks (95% CIs) for current smokers compared with never-smokers after adjustment for cardiovascular risk factors and public health center were 1.27 (1.05 to 1.54) for total stroke, 0.72 (0.49 to 1.07) for intraparenchymal hemorrhage, 3.60 (1.62 to 8.01) for subarachnoid hemorrhage, and 1.66 (1.25 to 2.20) for ischemic stroke. The respective multivariate relative risks among women were 1.98 (1.42 to 2.77), 1.53 (0.86 to 4.25), 2.70 (1.45 to 5.02), and 1.57 (0.86 to 2.87). There was a dose-response relation between the number of cigarettes smoked and risks of ischemic stroke for men. A similar positive association was observed between smoking and risks of lacunar infarction and large-artery occlusive infarction, but not embolic infarction. CONCLUSIONS: Smoking raises risks of total stroke and subarachnoid hemorrhage for both men and women and risk of ischemic stroke, either lacunar or large-artery occlusive infarction, for men. PMID- 15118171 TI - Impact of surgical clipping on survival in unruptured and ruptured cerebral aneurysms: a population-based study. AB - BACKGROUND AND PURPOSE: The management of aneurysms is controversial because little is known about the impact of clipping on long-term outcome. This study was designed to evaluate long-term survival of patients with aneurysms undergoing clipping in a statewide population. METHODS: We used a retrospective design using an administrative database to identify patients hospitalized with aneurysms (1987 to 2001). Time-to-event analysis was used to determine the risk of death from all causes and from neurological causes. RESULTS: 4619 patients (mean age 54.7+/ 15.3, 66.3% female) were hospitalized with cerebral aneurysms. Survival among patients with ruptures was significantly lower compared with patients with unruptured aneurysm (P<0.001) with adjusted hazard ratio (HR) of death after clipping 40% higher (HR: 1.4; 95% CI: 1.2, 1.7) in patients with rupture compared with those that were unruptured. Survival estimates for unruptured patients who underwent clipping were significantly higher than among those unruptured patients who did not undergo clipping (P<0.001), with adjusted HR of death 30% higher in patients with unruptured aneurysm that were not clipped compared with unruptured patients who were clipped (HR: 1.3; 95% CI: 1.1, 1.6). Patients with unruptured aneurysm who underwent clipping and survived beyond the 30-day postoperative period were less likely to die from neurologically related causes (5.6 versus 2.3%, P<0.001). Patients with ruptures and aneurysms who underwent clipping have a higher rate of death compared with the general population in the long-term. CONCLUSIONS: Short-term and long-term mortality after clipping of cerebral aneurysms is higher than previously reported. Patients with unruptured aneurysms who undergo clipping have improved survival compared with those who do not undergo clipping. This study supports the use of early intervention in the management of patients with unruptured aneurysms. PMID- 15118172 TI - Recurrent stroke in patients with symptomatic carotid artery occlusion is associated with high-volume flow to the brain and increased collateral circulation. AB - BACKGROUND AND PURPOSE: To investigate whether the risk of recurrent ipsilateral ischemic stroke in patients with symptomatic carotid artery occlusion (CAO) is related to (1) volume flow in the contralateral internal carotid artery (ICA), basilar artery (BA), and middle cerebral arteries (MCAs), and (2) intracranial collateral flow to the symptomatic side, measured in the first 6 months after the qualifying symptoms occurred. METHODS: We prospectively studied 112 patients with symptomatic CAO. Quantitative volume flow was measured with magnetic resonance angiography (MRA) and collateral flow via the circle of Willis with MRA, via the ophthalmic artery (OA) with transcranial Doppler sonography, and via leptomeningeal anastomoses with conventional angiography. RESULTS: During 49+/-14 months of follow-up (mean+/-SD), 7 patients had recurrent ipsilateral ischemic stroke. Compared with patients without recurrent stroke, these patients had significantly higher total flow to the brain, ie, ICA+BA flow (mean 536 mL/min versus 410 mL/min; P<0.05), and significantly higher contralateral ICA flow (355 mL/min versus 209 mL/min; P<0.001), whereas BA and MCA flow showed no significant differences. Also, they more often had Willisian collateral flow (P<0.05), mainly caused by increased collateral flow via the posterior communicating artery (PCoA; 71% versus 28%; P<0.05), whereas collateral flow via the OA and leptomeningeal anastomoses did not differ significantly. CONCLUSIONS: Recurrent ipsilateral ischemic stroke in patients with symptomatic CAO is associated with high volume flow to the brain and increased collateral PCoA flow. PMID- 15118173 TI - Agreement between ambulance paramedic- and physician-recorded neurological signs with Face Arm Speech Test (FAST) in acute stroke patients. AB - BACKGROUND AND PURPOSE: Patients with suspected stroke first assessed by ambulance paramedics require early recognition to facilitate appropriate triage and early treatment. We determined paramedic's accuracy in detecting acute stroke signs by comparing agreement between neurological signs recorded in the Face Arm Speech Test (FAST), a stroke recognition instrument, by paramedics on the scene and by stroke physicians after admission. METHODS: Suspected stroke patients admitted by ambulance paramedics directly to an acute stroke unit through a rapid ambulance protocol were examined by a trainee stroke neurologist or admitting stroke physician over a 1-year period. Recorded neurological signs (facial weakness, arm weakness, speech disturbance) in confirmed acute stroke/transient ischemic attack (TIA) cases were compared between paramedics and the stroke neurologist/physician. RESULTS: Ambulance crews referred 278 suspected stroke patients of whom 217 (78%) had confirmed stroke (n=189) or TIA (n=28); 95% were examined by the stroke neurologist (median 18 hours after paramedic assessment). Recorded signs and agreement between paramedics and stroke physicians in confirmed stroke group were: facial weakness, 68% versus 70% (kappa=0.49; 95% CI: 0.36 to 0.62); arm weakness, 96% versus 95% (kappa=0.77; 95% CI: 0.55 to 0.99); and speech disturbance, 79% versus 77% (kappa=0.69; 95% CI: 0.56 to 0.82). Interrater agreement was complete for arm weakness in 98% cases. CONCLUSIONS: Recognition of neurological deficits by ambulance paramedics using FAST shows good agreement with physician assessment, even allowing for temporal evolution of deficits. The high prevalence and good agreement for arm weakness suggest that this sign may have the greatest usefulness for prehospital ambulance triage and paramedic-based neuroprotective trials. PMID- 15118174 TI - Internal carotid artery occlusive disease and polymorphisms of fractalkine receptor CX3CR1: a genetic risk factor. AB - BACKGROUND AND PURPOSE: Fractalkine (FKN), a chemokine expressed by inflamed endothelium, induces leukocyte adhesion and migration via the receptor CX3CR1. The polymorphisms V249I and T280M affect receptor expression and function. The role of FKN in atherosclerosis has been recently demonstrated. The aim of this study was to investigate a possible association between CX3CR1 polymorphisms and increased risk of internal carotid artery (ICA) occlusive disease. METHODS: We studied 108 patients consecutively recruited for ICA occlusive disease, 84 of whom underwent operation for carotid endarterectomy, and 204 subjects without ICA occlusive disease (controls). Polymorphic genotypes were determined by polymerase chain reaction and sequencing analysis. RESULTS: The adjusted odds ratio (OR) associated with the presence of the M280 (TM+MM versus TT genotype) was 0.55 (95% CI: 0.29 to 0.99; P=0.037). Therefore, this allele is associated with a reduced risk of ICA occlusive disease. No significant differences were observed in I249 distribution. The frequency of I249 allele was significantly higher in cases of hard plaques, which are considered more stable than soft ones (OR: 0.38; 95% CI: 0.13 to 1.05; P=0.037). Multiple logistic regression analysis using the common risk factors and the I249 and M280 allele variants revealed that the M280 allele was an independent risk factor for ICA stenosis (P=0.047). CONCLUSIONS: The results show that the CX3CR1 M280 is an independent genetic risk factor for ICA occlusive disease and that I249 is involved in the stability of carotid plaques. Even if obtained from a relatively limited patient series, these results might have relevant implications for treatment of ICA stenosis and possibly prevention of carotid related stroke. Further prospective cross-sectional studies are needed to confirm these results. PMID- 15118175 TI - Cerebral blood flow and metabolism during infusion of norepinephrine and propofol in patients with bacterial meningitis. AB - BACKGROUND AND PURPOSE: In patients with severe bacterial meningitis, norepinephrine is often infused to increase mean arterial pressure (MAP). This increases cerebral blood flow (CBF), but it is unknown if this increase is caused by impaired cerebral autoregulation or by a cerebral effect of norepinephrine through increased cerebral metabolism. The latter possibility implies a CBF metabolism coupling. This has not been studied during meningitis. We studied the effect of norepinephrine and propofol on CBF and oxidative metabolism in patients with severe bacterial meningitis. METHODS: In seven patients with pneumococcal meningitis and 7 healthy subjects, norepinephrine was infused intravenously; patients also underwent intravenous propofol infusion. Global CBF was measured by the Kety-Schmidt technique; cerebral oxidative metabolism and net flux of norepinephrine and epinephrine were calculated from measured arterial-to-jugular venous concentration differences (a-vD). RESULTS: During norepinephrine infusion, MAP increased from a median value of 79 (range, 70 to 89) to 99 (98 to 129) mm Hg in patients, and from 87 (72 to 103) to 123 (112 to 132) mm Hg in controls. CBF increased in patients (51 [48 to 60] to 59 [54 to 77] mL/100 g per minute) but remained unchanged in controls. The cerebral metabolic rate of oxygen (CMRO2) decreased in patients and remained unchanged in controls. No cerebral net flux of norepinephrine or epinephrine was found at any time in the 2 groups. During propofol infusion, CMRO2, and the a-vDO2 decreased whereas CBF was unchanged. CONCLUSIONS: In patients with severe bacterial meningitis, norepinephrine increases both MAP and CBF but not CMRO2, indicating impaired autoregulation. Propofol reduces CBF relatively less than cerebral metabolism, suggesting a resetting of the CBF-CMRO(2) relationship. PMID- 15118176 TI - Effects of urban community intervention on 3-year survival and recurrence after first-ever stroke. AB - BACKGROUND AND PURPOSE: For the past 2 decades, stroke has been a principal cause of death in China, and stroke incidence tends to increase with the increase of stroke-related risk factors. The purpose of this study was to evaluate the effects of urban community-based intervention on 3-year survival and recurrence after first-ever stroke. METHODS: Two communities with a registered population of approximately 50 000 each were selected as either intervention or control communities in Beijing during 1991 to 2000. Comprehensive intervention measures including the management of high-risk population and the health education of whole community population were regularly implemented. Then the influence of community intervention on 3-year survival and recurrence after initial stroke was evaluated. RESULTS: Within 3 years, 41.85% of 736 patients in the intervention community died whereas 40.34% of 818 patients in the control community died. Of 223 cases from the intervention community, 26 (11.66%) had a recurrent stroke within 3 years versus 52 (20.80%) of 250 cases from the control community. The statistical difference was found. Compared with the control community, the death risk of first-ever stroke in the intervention community decreased by 26% (relative risk [RR]=0.74; 95% confidence interval [CI]: 0.61 to 0.89; P=0.002); especially, that of hemorrhagic stroke decreased by 39% (RR=0.61; 95%CI: 0.46 to 0.81; P=0.001). Compared with the control community, the recurrence risk of first ever stroke from the intervention community decreased by 42% (RR=0.58; 95% CI: 0.34 to 1.00; P=0.048). CONCLUSIONS: Community intervention may be effective and beneficial to the recurrence prevention and survival improvement of stroke, especially hemorrhagic stroke. PMID- 15118177 TI - Oxidative stress is associated with XIAP and Smac/DIABLO signaling pathways in mouse brains after transient focal cerebral ischemia. AB - BACKGROUND AND PURPOSE: The interaction of X chromosome-linked inhibitor-of apoptosis protein (XIAP) with second mitochondria-derived activator of caspase (Smac)/direct inhibitor-of-apoptosis protein-binding protein with low pI (DIABLO) contributes to regulation of apoptosis after a variety of cell death stimuli, and in our reported in vivo transient focal cerebral ischemia (tFCI) model. We have also reported that overexpression of copper/zinc superoxide dismutase (SOD1) reduces apoptotic cell death after tFCI. Our present study was designed to clarify the relationship between the XIAP signaling pathway and oxidative stress in the regulation of apoptosis after tFCI. METHODS: We used a tFCI model of SOD1 transgenic mice and wild-type littermates to examine the expression of XIAP and Smac/DIABLO by Western blotting and the interaction of XIAP with Smac/DIABLO (XIAP/Smac) or caspase-9 (XIAP/caspase-9) by coimmunoprecipitation. The direct oxidation of carbonyl groups, an indication of oxidative injury to total and individual proteins caused by tFCI, was examined using a 2,4 dinitrophenylhydrazone reaction assay. RESULTS: Direct oxidative injury to cytosolic and mitochondrial proteins was reduced by SOD1 after tFCI. The individual oxidized carbonyls in XIAP, mitochondrial Smac/DIABLO, and caspase-9 were also reduced by SOD1. Expression of XIAP and XIAP/caspase-9 was promoted, whereas translocation of Smac/DIABLO and XIAP/Smac was reduced, by SOD1 after tFCI. CONCLUSIONS: These results suggest that overexpression of SOD1 may affect the XIAP pathway after tFCI by reducing the direct oxidative reaction to XIAP regulators after reperfusion injury. PMID- 15118178 TI - Critical closing pressure in subarachnoid hemorrhage: effect of cerebral vasospasm and limitations of a transcranial Doppler-derived estimation. AB - BACKGROUND AND PURPOSE: Critical closing pressure (CCP) is thought to be jointly influenced by intracranial pressure and cerebrovascular tone. We examined how CCP is affected by cerebral vasospasm after subarachnoid hemorrhage (SAH). METHODS: In 15 patients with vasospasm of the middle cerebral artery, CCP was calculated using 2 methods previously reported (ad modem Aaslid and Michel, indexed CCP(Aaslid) and CCP(Michel), respectively) based on data of arterial blood pressure and flow velocity (FV) as assessed by transcranial Doppler. RESULTS: CCP decreased significantly (P<0.05) during vasospasm (CCP(Aaslid)=6.3+/-22.9 mm Hg, CCP(Michel)=14.9+/-16.5 mm Hg, mean+/-SD) as compared with baseline (CCP(Aaslid)=24.4+/-20.3 mm Hg, CCP(Michel)=27.8+/-19.4 mm Hg). This was not attributable to ICP, which remained unaffected by vasospasm. In addition, CCP was significantly lower on the side of vasospasm (CCP(Aaslid)=11.9+/-24.2 mm Hg, CCP(Michel)=18.4+/-19.6 mm Hg) as compared with the contralateral nonvasospastic side (CCP(Aaslid)=24.7+/-22.3 mm Hg, CCP(Michel)=28.2+/-18.0 mm Hg). CONCLUSIONS: Assuming that autoregulation-related distal vasodilatation outweighs proximal vasospasm, CCP should decrease. Alternatively, CCP might have increased during vasospasm as the tension of big vessels increase, but the turbulence occurring during vasospasm may have impaired the linear relationship between pressure and FV, thus leading to a marked underestimation of CCP. In conclusion, interpretation of CCP in vasospasm is difficult and may be overshadowed by nonlinear hemodynamic effects. PMID- 15118179 TI - Aortic arch atherosclerotic lesions and the recurrence of ischemic stroke. AB - BACKGROUND AND PURPOSE: Aortic arch atherosclerotic lesions are often associated with embolic brain infarction. We investigated the relationship between stroke recurrence and the characteristics of aortic arch atherosclerotic lesions. METHODS: Among 487 stroke patients who underwent transesophageal echocardiography, 283 patients with brain embolism diagnosed without significant occlusive lesions (> or =50%) in their cerebral arteries were included in this study. We measured the intima-media thickness (IMT) and evaluated the extension and mobility of the aortic arch atherosclerotic lesions. During a mean follow-up period of 3.4 years, we investigated the relationship between stroke recurrence and the various characteristics of the aortic arch atherosclerotic lesions. RESULTS: An IMT > or =4.0 mm was found in 67 patients (25.3%). In 51 of these patients, the aortic lesions extended to the origin of the branches of the arch. Recurrences of cerebral ischemic events were found in 32 patients (recurrence group) and not in the other 251 (nonrecurrence group). Aortic atheroma > or =4.0 mm (41% versus 22%), aortic atheroma extending to the branches (63% versus 39%), and both (38% versus 16%) were more frequently seen in the recurrence group than in the nonrecurrence group (P<0.05, P<0.1, P<0.01, respectively). After adjustment for age and the presence of hypertension, an aortic atheroma that was > or =4.0 mm as well as extending to the branches was found to be an independent predictor of ischemic stroke recurrence (hazard ratio=2.42, P<0.05). CONCLUSIONS: Stroke recurrence is associated with the severity of the atheroma (IMT > or =4.0 mm) and plaque extension to the branches. PMID- 15118180 TI - Role of p38 mitogen-activated protein kinase on cerebral vasospasm after subarachnoid hemorrhage. AB - BACKGROUND AND PURPOSE: Inflammatory cytokines are involved in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH). This study was conducted to examine the role of p38 mitogen-activated protein kinase (MAPK) in the development of vasospasm and cytokine production. METHODS: We measured the expression levels of genes and proteins related to inflammation in human vascular smooth muscle cells (hVSMCs) treated with hemolysate and FR167653 (FR) (1 micromol/L), a selective p38MAPK inhibitor, for 48 hours by TaqMan real-time reverse transcription-polymerase chain reaction (RT-PCR) and ELISA. Twenty-one dogs were assigned to 3 groups of 7 animals: control, placebo, and FR-treated (1 mg/kg/d) groups in a double-hemorrhage model. The effects were assessed through the caliber of the basilar artery, and the changes in gene expressions and the activation of p38MAPK were assessed by Western blot analysis. RESULTS: Treatment of hVSMCs with hemolysate induced significant upregulation of interleukin (IL) 1alpha, IL-1beta, and IL-8 gene and protein expressions, which was suppressed significantly with FR. The mean vessel caliber on day 7, as a percentage of that of day 0, was 49% in the placebo, and 74% in the FR group (P=0.0001). The gene expression levels of IL-1alpha, IL-1beta, and IL-8 in the arterial wall were extremely elevated in the placebo, and significantly suppressed in the FR group (P=0.0027, 0.0002, and 0.0073). p38MAPK phosphorylation was stimulated in the placebo and hemolysate in vitro, and suppressed in the FR group. CONCLUSIONS: These results suggest that p38MAPK is activated in the arterial wall after SAH, leading to the development of vasospasm, possibly through the upregulation of inflammatory cytokines. PMID- 15118181 TI - Predictive values of lacunar transient ischemic attacks. AB - BACKGROUND AND PURPOSE: We postulated that a lacunar syndrome occurring with transient ischemic attacks (TIAs) or progressive nonsudden onset predicts a brain infarction (BI), presumably caused by a small artery disease (ie, lacunar BI) better than a lacunar syndrome with sudden onset. METHODS: We included 510 patients with BI. BI was classified into etiologic groups including lacunar BI group. We identified the patients with lacunar or nonlacunar syndrome, and those with TIAs preceding the BI or with symptoms of nonsudden onset. RESULTS: Nonlacunar syndrome had a negative predictive value for a lacunar BI of 95%. A lacunar syndrome had a positive predictive value (PPV) of 57% for lacunar infarction (n=109), and the PPV increased to 79% in the case of recent TIAs preceding the lacunar syndrome. Hypertension was present in 95% of cases with lacunar TIAs (odds ratio: 10.69; 95% confidence interval: 1.34 to 84.82; P=0.02). CONCLUSIONS: Lacunar TIAs are almost always associated with history of arterial hypertension and have a high PPV for lacunar BI. This subgroup of patients may reflect different underlying mechanisms than the group of patient with lacunar syndrome of sudden onset. PMID- 15118182 TI - Health care resource use after acute stroke in the Glycine Antagonist in Neuroprotection (GAIN) Americas trial. AB - BACKGROUND AND PURPOSE: To compare 3-month stroke outcomes and stroke-related health care resource use between the US and Canada in the Glycine Antagonist in Neuroprotection (GAIN) Americas study. Delivery of medical care for stroke patients, often driven by efforts to curb costs, varies substantially between countries. Data on the potential impact of these variations on clinical outcomes are sparse. METHODS: The analysis of health care resource included total length of stay (LOS) in hospital, intensive care unit (ICU), and acute-care ward or rehabilitation unit, or both; number of outpatient rehabilitation sessions and visits to a physician; place of residence after discharge; and employment status. Cox proportional hazards models and logistic regression were used to calculate survival hazards and predictors of favorable functional outcome (Barthel Index of 95 to 100). RESULTS: One thousand six hundred four patients who were independent before stroke (mean age: 69.9+/-12.7 years, 53% men, 85% ischemic stroke, 69% in the US) were included. Three-month survival and functional outcome did not differ between the US and Canada. Survival rate was 80% in both countries. Favorable functional outcome was achieved in 43% of Canadian and 47% of US patients. Fewer Canadian patients received treatment in ICU (19% versus 63% in the US), and Canadians had longer stays in hospital or rehabilitation facility (median: 33 days versus 16 days in the US). CONCLUSIONS: Despite similar 3-month survival and functional outcome, patterns of health care resource varied substantially between the US and Canada. US patients had more intensive early care; Canadian patients had longer hospitalizations and rehabilitation care. Further research is required to determine the most cost-effective treatment and rehabilitation plan for people who have a stroke. PMID- 15118184 TI - Long-term relative survival in elderly patients after carotid endarterectomy. PMID- 15118185 TI - Stroke is best managed by neurologists. PMID- 15118186 TI - White matter hyperintensities are associated with impairment of memory, attention, and global cognitive performance in older stroke patients. AB - BACKGROUND: The importance of white matter hyperintensities (WMH) for cognitive performance in older stroke patients is largely unknown. We hypothesized that processing speed and executive dysfunction will be associated with frontal WMH whereas impaired memory will be associated with temporal WMH. METHODS: Neuropsychological assessments using the Cambridge Cognitive Examination (CAMCOG) and the Cognitive Drug Research (CDR) were completed for 96 stroke survivors aged older than 75 and 23 age-matched controls. Magnetic resonance imaging whole-brain axial FLAIR images were undertaken to visualize WMH and an automated threshold technique was used to determine their volume. RESULTS: In comparison to controls, the stroke patients had significantly greater volume of WMH in all key areas. Within the stroke group, a consistent pattern of significant association was identified between total and frontal WHM volumes and attention and processing speed tasks (eg, choice reaction time [right: R=0.24 P=0.02; left: R=0.26, P=0.01]), but not with executive function. There were significant associations between memory and temporal WMH volumes (right: R=0.27, P=0.008; left: R=0.20, P=0.047). CONCLUSIONS: In older stroke patients, cognitive processing speed and performance on measures of attention are significantly associated with WMH volume, particularly in the frontal lobe regions, whereas memory impairment is associated with the volume of temporal lobe WMH. PMID- 15118187 TI - Moderate intensity resistance exercise, plus or minus soy intake: effects on serum lipid peroxides in young adult males. AB - Lipid peroxides can be both a product and an initiator of oxidant stress. Conceivably, exercise can either increase concentrations of lipid peroxides (by causing oxidant stress), or decrease them (by accelerating peroxide breakdown). The net effect could depend on exercise intensity and nutritional intake of antioxidants. The present study examined the response of serum lipid peroxides to the combination of moderate intensity, weight resistance exercise plus intake of soy protein, a source of antioxidant phytochemicals. Recreationally trained, young adult men (N = 18) consumed soy protein or antioxidant-poor whey protein for 4 weeks (40 g protein/d) before a session of moderate intensity, weight resistance exercise. In the soy group, exercise decreased values for serum lipid peroxides at 5 min, 3 h, and 24 h post-exercise. The whey group showed the depression only at 24 h. In both the soy and whey groups, a small rise was seen for interleukin-8, which is consistent with the idea that the exercise session induced a moderate muscle stress. In summary, a moderate intensity, weight resistance exercise session, despite inducing mild inflammation, depressed plasma serum peroxide values, especially when combined with 4 weeks of soy consumption. PMID- 15118188 TI - Effects of vitamin-mineral supplementation on cardiac marker and radical scavenging enzymes, and MDA levels in young swimmers. AB - The relationship among the enzyme activities of cardiac markers, the antioxidant defense system, and erythrocyte membrane malonyldialdehyde (MDA) levels related to vitamin-mineral supplementation in swim exercise was investigated. Swimmers aged 11 - 13 years were divided into 2 separate groups as control and vitamin mineral supplemented. Swimmers participated in a monthly swimming program (4 times/wk) and swam approximately 2- 2.5 km/d. Cardiac markers such as creatine kinase (CK), creatine kinase-MB (CK - MB), glutamic oxaloacetic transaminase [GOT (AST)], lactate dehydrogenase (LDH), and anti-oxidant enzymes such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activities in post-training samples were found to be significantly (p<.05) higher than in pre-training samples. Except for GOT (AST), the activity increases in CK, CK-MB, and LDH in female and male supplemented groups were significantly (p<.05) lower than those of control groups during the 1-month period of swim training. Antioxidant enzyme activity increases in the male vitamin-mineral group were significantly (p <.05) higher when compared with the other groups. Post-training MDA levels were significantly (p <.001) higher than pre-training MDA levels in the control groups, whereas no significant (p<.05) differences were found between the vitamin-mineral supplemented groups. Vitamin-mineral supplementation was found to attenuate cardiac and muscle damage markers while also enhancing antioxidant levels and reducing membrane LPO levels in response to 1 month of swim training. PMID- 15118189 TI - Antioxidant diet supplementation influences blood iron status in endurance athletes. AB - OBJECTIVE: The aim of this work was to check the effects of antioxidant supplementation (vitamins E and C, and beta-carotene) on the basal iron status of athletes prior to and following their training and competition season (3 months). DESIGN: Eighteen amateur trained male athletes were randomly distributed in 2 groups: placebo (lactose) and antioxidant supplemented (vitamin E, 500 mg/d; vitamin C, 1 g/d; and beta -carotene, 30 mg/d). The study was double blind. Hematological parameters, dietary intake, physical activity intensity, antioxidant status (GSH/GSSG ratio), and basal iron status (serum iron, transferrin, ferritin, and iron saturation index) were determined before and after the intervention trials. RESULTS: Exercise decreased antioxidant defenses in the placebo group but not in the antioxidant-supplemented group. No changes were found in the number of erythrocytes, hematocrit, or hemoglobin concentration, or in values of serum iron parameters, after taking the antioxidant cocktail for 3 months, in spite of the exercise completed. The placebo group showed a high oxidative stress index, and decreases in serum iron (24%) and iron saturation index (28%), which can neither be attributed to aspects of the athletes' usual diet, nor to hemoconcentration. CONCLUSIONS: Antioxidant supplementation prevents the decrease of serum iron and the iron saturation index, and a link between iron metabolism and oxidative stress may also be suggested. PMID- 15118190 TI - Reduced neuromuscular activity with carbohydrate ingestion during constant load cycling. AB - The purpose of this study was to determine the effect of carbohydrate ingestion before and during intense constant load cycling to volitional fatigue on surface electromyographic (sEMG) activity from the vastus lateralis (VL) and vastus medialis (VM) muscles. After 24-h diet and training control, 8 well-trained subjects (maximal O(2 )uptake (VO(2max )) 66+/- 2 ml. kg-1. min-1; mean plusmn; SD) ingested 8 ml. kg-1 of either a 6.4% carbohydrate-electrolyte (CHO) or a placebo (PLA) solution immediately before, followed by 2 ml. kg-1 of the same solution every 15 min while cycling to exhaustion at 84+/- 1% of VO(2max.) Exercise time to fatigue was 13% longer with CHO ingestion compared to PLA (58:54+/- 8:48 vs. 51: 18 +/- 5:54 min:s, NS). VO(2 ) (4.22 +/- 0.11 vs. 4.20 +/- 0.14 L. min-1 ), heart rate (172 +/- 4 vs. 176 +/- 4 beats. min-1), ratings of perceived effort (18 +/- 0.1 vs. 19+/- 0.1), and rates of carbohydrate oxidation (314 +/- 28 vs. 324 +/- 26 ?mol. kg-1. min -1) were similar for both PLA and CHO at exhaustion. There was no main treatment effect of CHO ingestion on blood glucose or lactate concentrations, nor plasma prolactin levels either during exercise or at fatigue. However, CHO ingestion attenuated the rise in EMG root mean square (RMS) activity during the latter stages (>45 min) of exercise and at the point of exhaustion for both VM (0.325 +/- 0.010 vs. 0.403 +/- 0.020 mV; =p.006) and VL (0.298+/- 0.011 vs. 0.370 +/- 0.007 mV; p =.0004). We conclude that in well-trained subjects, the ingestion of carbohydrate attenuated the increase in surface electromyographic activity during intense, constant load cycling leading to exhaustion in approximately 1 h. The precise mechanism(s) underlying this effect cannot be attributed to alterations in CHO availability but, instead, may be linked to changes in afferent sensory input. PMID- 15118191 TI - Inadequate carbohydrate intake following prolonged exercise does not increase muscle soreness after 15 minutes of downhill running. AB - In Delayed Onset Muscle Soreness (DOMS), muscles become sore 24 to 48 hours after eccentric and unaccustomed activity. Fiber stiffness, due to decreased muscle glycogen, may predispose muscle to greater damage during eccentric exercise. This study sought to determine if inadequate carbohydrate intake following a protocol to decrease muscle glycogen would increase DOMS after 15 min of downhill running. Thirty-three male subjects (age, 18-35 years) were randomized into 3 groups for testing over a 7-day period. The depletion (DEP) group (n= 12) underwent a glycogen depletion protocol prior to a 15-min downhill run designed to induce DOMS. The repletion (FED) group (n = 10) underwent a glycogen depletion protocol followed by a carbohydrate repletion protocol (>80% CHO) prior to downhill running. The third (ECC) group (n = 11) performed only the downhill running protocol. Subjective muscle soreness, isometric force production, relaxed knee angle, and thigh circumference were measured pretreatment and on days 1, 2, 3, 4, and 6 post treatment. Subjective muscle soreness for all groups increased from 0 cm pretreatment to 3.05 +/- 0.72 cm (on a 10-cm scale) on day 1 post treatment (p<.05). All groups were significantly different from baseline measurements until day 4 post treatment. Each group experienced a decline in isometric force from 281 +/- 45 N pre- to 253 +/- 13 N on day 1 post treatment (p <.05). The decrease in isometric force persisted in all groups for 4 days post treatment. Increases in thigh circumference and relaxed knee angle elevations in all 3 groups were statistically different (p <.05) from pretreatment until day 4. No differences were noted between groups for any of the parameters examined. In the current study, 15 min of downhill running is sufficient to cause DOMS with the associated functional and morphological changes; however, inadequate carbohydrate intake after a glycogen depleting exercise does not appear to exacerbate DOMS and the associated symptoms. PMID- 15118192 TI - Effect of active versus passive recovery on metabolism and performance during subsequent exercise. AB - This study tested the hypothesis that active recovery between bouts of intense aerobic exercise would lead to better maintenance of exercise performance in the second bout of exercise. Seven trained men on 2 separate occasions (VO(2peak) = 58.3+/- 9.4 ml x kg(-1) x min(-1)) performed as much work as possible during two 20-min cycling exercise bouts, separated by a 15-min recovery period. During passive recovery (PR), subjects rested supine, while during active recovery (AR) subjects continued to cycle at 40% VO(2peak). Muscle biopsies and blood samples were obtained. Neither muscle glycogen or lactate was different when comparing AR with PR at any point. In contrast, plasma lactate concentration was higher (p<.05) in PR versus AR during the recovery period, such that subjects commenced the second bout of intense exercise with a lower (p <.05) plasma lactate concentration in AR (4.4 +/- 0.7 vs. 7.7 +/- 1.4 mmol. L(-1) following AR and PR, respectively). Work performed in Bout 2 was less than that performed in Bout 1 in both trials (p<.01), with no difference in work performed between trials. These data do not support the benefit of AR when compared to PR in the maintenance of subsequent intense aerobic exercise performance. PMID- 15118193 TI - Effect of ingested fluid composition on exercise-related transient abdominal pain. AB - The present study investigated the effect of ingested fluid composition on the experience of exercise-related transient abdominal pain (ETAP). Forty subjects, susceptible to ETAP, completed 4 treadmill exercise trials: a no-fluid trial and flavored water (FW, no carbohydrate, osmolality = 48 mosmol/L, pH = 3.3), sports drink (SD, freshly mixed Gatorade, 6% total carbohydrate, 295 mosmol/L, pH = 3.3), and reconstituted fruit juice (FJ, BERRI trade mark orange, 10.4 % total carbohydrate, 489 mosmol/L, pH= 3.2) trials. Measures of the experience of ETAP and gastrointestinal disturbances, particularly bloating, were quantified. The FJ was significantly (p =.01) more provocative of both ETAP and bloating than all other trials. There was no difference among the no-fluid, FW, and SD in the severity of ETAP experienced, although the difference between the no-fluid and SD approached significance at the.05 level (p =.056). There was a significant relationship between both the mean (r = 0.40, p =.01) and peak (r= 0.44, p=.01) levels of ETAP and bloating. When the level of bloating was controlled for, the FJ remained significantly (p =.01) more provocative of ETAP than the other conditions, with no difference between the FW and SD (p =.37). The results indicate that in order to avoid ETAP, susceptible individuals should refrain from consuming reconstituted fruit juices and beverages similarly high in carbohydrate content and osmolality, shortly before and during exercise. Further, the mechanism responsible for the heightened experience of ETAP in the FJ trial extends beyond a gastric mass explanation. PMID- 15118195 TI - Evaluation of nutrition knowledge and dietary recommendations by coaches of adolescent Brazilian athletes. AB - The objectives of this study were to describe the dietary practices recommended by coaches working with adolescent athletes and to assess their nutritional knowledge. During a regional competition in the state of Sao Paulo, Brazil, 55 coaches were interviewed. These coaches represented 22 cities with athletes enrolled in Olympic gymnastics, tennis, swimming, and judo events. A 3-section questionnaire was used to obtain data on demographic characteristics, dietary recommendations, and nutrition knowledge. Results showed that all coaches recommended general dietary practices during training, with no specific strategies for pre-, during-, and post-training periods. The main objectives of the recommendations for the training period were weight control and muscle mass gain. Deleterious weight control practices were recommended by 27% of the coaches. Specific dietary practices pre and post competition were recommended by 93% and 46% of the coaches, respectively. Participants responded correctly to 70% (SD = 3.2) of the nutrition knowledge questions, with no significant differences (p =.61) between sports. The knowledge test identified a tendency to over-value proteins, excessively low-fat diets, and food myths. These findings indicate the importance of developing strategies that will enhance the nutritional training of coaches. PMID- 15118194 TI - Reliability and validity of a brief questionnaire to assess calcium intake in female collegiate athletes. AB - Calcium intake often is inadequate in female collegiate athletes, increasing the risk for training injuries and future osteoporosis. Thus, a brief and accurate assessment tool to quickly measure calcium intake in athletes is needed. We evaluated the reliability and validity, compared to 6 days of diet records (DRs), of the Rapid Assessment Method (RAM), a self-administered calcium checklist. Seventy-six female collegiate athletes (mean age = 18.8 yrs, range= 17- 21; 97 % Caucasian) were recruited from basketball, cross-country, field hockey, soccer, and volleyball teams. Athletes completed a RAM at the start of the training season to assess calcium intake during the past week. Two weeks later, a second RAM was completed to assess reliability, and athletes began 6 days of diet records (DRs) collection. At completion of DRs, athletes completed a final RAM, corresponding to the same time period as DRs, to assess agreement between the 2 instruments. The RAM demonstrated adequate test-retest reliability over 2 weeks (n= 56; Intraclass correlation [ICC] = 0.54, p < 0.0001) and adequate agreement with DRs (n = 34; ICC = 0.41, p = 0.0067). Calcium intake was below recommended levels, and mean estimates did not differ significantly on the RAM (823 +/- 387 mg/d) and DRs (822 +/- 330 mg/d; p = 0.988). Adequacy of calcium intake from both DRs and the RAM was classified as "inadequate" (<1000 mg/d) and "adequate" (> or = 1000 mg/d). Agreement between the RAM and DRs for adequacy classification was fair (ICC= 0.30, p = 0.042), with the RAM identifying 84% of athletes judged to have inadequate calcium intake based on DRs. The RAM briefly and accurately estimates calcium intake in female collegiate athletes compared to DRs. PMID- 15118196 TI - Cordyceps Sinensis (CordyMax Cs-4) supplementation does not improve endurance exercise performance. AB - It is purported that supplementation with Cordyceps Sinensis (CordyMax Cs-4) will improve oxidative capacity and endurance performance. The intent of this investigation was to examine the effects of CordyMax Cs-4 supplementation on VO<(2peak,) ventilatory threshold, and endurance performance in endurance-trained cyclists. Twenty-two male cyclists participated in 5 weeks of supplementation with CordyMax Cs-4 tablets (3 g/d). Training intensity was maintained by weekly documentation and reporting throughout the 5-week period. Subjects completed a VO(2peak) test and work-based time trial prior to and following the supplementation period. VO(2peak) was similar within and between placebo (PLA) and treatment (CS) groups prior to (59.9 +/- 5.9 vs. 59.1 +/- 5.4 ml/kg/min, respectively) and following (60.1 +/- 5.5 vs. 57.1 +/- 5.8 ml/kg/min, respectively) the supplementation period. Ventilatory threshold (VT) was measured at 72 +/- 10% of VO(2peak) in P and T prior to supplementation and did not change in either group following the supplementation. PLA completed the time trial in 61.4+/- 2.4 min compared to 62.1+/- 4.0 min in T. Time trial measurements did not differ between groups, nor did they change in response to supplementation. It is concluded that 5 weeks of CordyMax Cs-4 supplementation has no effect on aerobic capacity or endurance exercise performance in endurance-trained male cyclists. PMID- 15118197 TI - Bimanual coordination: an unbalanced field of research. AB - Using more than one limb to perform functional, goal-directed actions is arguably one of the most important abilities that human beings possess. In many everyday tasks, the hands, in particular, must be used to accomplish all manner of goals. From buttoning a shirt to opening a jam jar and driving to work, good bimanual coordination is of great utility. In addition to the tasks mentioned above, there are also other tasks involving the functional use of more than one limb, including walking or cycling and typing a report. With a little thought, it becomes apparent that there is at least one important difference between these categories of coordination tasks. On one hand, in some tasks the effectors must perform markedly different motor outputs that are bound together in some functionally defined and usually object-oriented manner (e.g., buttoning a shirt) yet, in others, the effectors produce very similar motor outputs but in a specific temporal order, which may or may not repeat itself periodically (e.g., walking and cycling compared to typing or drumming). In this short article, I will argue that the second category of coordination task and, in particular, cyclical coordination, has been studied extensively and, at least at the level of behavior, is relatively well understood. In contrast the former category of bimanual task is seldom studied and, even at the descriptive level, is rather poorly understood. One of the reasons for this may be the complexity of such tasks and the technical difficulties involved in attempting to study them. By highlighting some key studies, I hope to illustrate that such tasks can be fruitfully studied in the laboratory. Last, since the neural control processes underlying both classes of coordination task are not yet well known, I aim to draw attention to the potential value of the interventional technique of Transcranial Magnetic Stimulation (TMS) as a tool for investigating the functions of brain regions contributing to bimanual coordination. PMID- 15118198 TI - Effect of movement termination in single- and dual-phase pointing tasks. AB - Two experiments are reported that focus on manipulating both the context and the spatial precision of a computer-pointing task. Single goal-directed actions are compared to dual-phase tasks, where participants are required to sequentially attain two goal locations. Results support the idea that for movements in series, movement planning, and online feedback, control can occur simultaneously. Additionally, for single-phase tasks and the final phase of dual-phase tasks, the termination requirement influences the temporal components of the movement. The effects of termination and movement context appear to hold regardless of the spatial precision of the task. This suggests that the effects of spatial precision and movement termination are independent, although both have an impact on the deceleration time for goal-directed movements. PMID- 15118199 TI - Disruption of coordination between arm, trunk, and center of pressure displacement in patients with hemiparesis. AB - To determine how arm movements influence postural sway in the upright position after stroke, interactions between arm, trunk, and center of pressure (CoP) displacements in the sagittal direction were investigated in participants with hemiparesis and healthy subjects. Participants swung both arms sagittally in either of 2 directions (in-phase, anti-phase) and at 2 speeds (preferred, fast) while standing on separate force plates. Variables measured included amplitude and frequency of arm swinging, shoulder and trunk range of motion, CoP displacements under each foot and of the whole body, and the relationships between the arm, trunk, and CoP displacements. CoP displacements under the non paretic leg were greater than those under the paretic leg, which may in part be related to the larger amplitude of swinging of the non-paretic arm. CoP displacements under each foot were not related to arm swinging during in-phase swinging at the preferred speed in healthy subjects. When speed of arm swinging was increased, however, the CoP moved in a direction opposite to the arm movement. In contrast, in individuals with hemiparesis, CoPs and arms moved in the same direction for both speeds. During anti-phase swinging in healthy subjects, the trunk counterbalanced the arm movements, while in participants with hemiparesis, the trunk moved with the affected arm. Results show that stroke resulted in abnormal patterns of arm-trunk-CoP interactions that may be related to a greater involvement of the trunk in arm transport, an altered pattern of coordination between arm and CoP displacements, and an impaired ability of the damaged nervous system to adapt postural synergies to changes in movement velocity. PMID- 15118200 TI - Adaptations in bilateral mechanical power patterns during obstacle avoidance reveal distinct control strategies for limb elevation versus limb progression. AB - The present study investigated the adaptations of specific power bursts during the combined contexts of the proximity (lead vs. trail limb) and height of an obstruction in relation to limb elevation versus progression. Ten young, adult, male subjects walked at their natural speed during unobstructed walking and the bilateral avoidance of moderate and high obstacles. Hip flexor generation power was unaffected by obstacle height for the leading limb and always delayed for the trailing limb. The knee extensor absorption power burst at toe-off was also eliminated for the trailing limb and was found to reappear in mid-swing. Few differences were seen for ankle push-off power. The results suggest that the hip joint is dedicated to limb advancement only, while the knee joint is directly involved in limb elevation and the control of multiarticular effects. PMID- 15118201 TI - Task-specific stabilization of postural coordination during stance on a beam. AB - Surfaces shorter in extent than the feet elicit multi-joint coordination that differs from what is elicited by stance on extensive surfaces. This well-known effect arises from the mechanics of the actor-environment interaction. Multi joint control of stance is also known to be influenced by non-mechanical aspects of a situation, including participants' task or intention. Intentional constraints do not originate in mechanics, and for this reason one might suppose that constraints imposed by mechanics would dominate constraints imposed by intentions, when the two were in conflict. We evaluated this hypothesis by varying participants' supra-postural task during stance on a short surface. While standing on a 10-cm wide beam, participants were exposed to optic flow generated by fore-aft oscillations of a moving room. Participants faced a target attached to the front wall of the moving room and were asked either to look at the target (with no instruction to move) or intentionally to track it with their head (i.e., to keep the target-head distance constant). Within trials, we varied the frequency of room (and target) motion, from 0.15 to 0.75 Hz, in steps of 0.05 Hz. In both conditions, ankle and hip rotations exhibited anti-phase coordination, but behavior was not identical across conditions. Coupling between motion of the room and the head was stronger for the tracking task than for the looking task, and the stability of ankle-hip coordination was greater during tracking than during looking. These results indicate that the influence of support surface mechanics did not eliminate the influence of the supra-postural task. Environment based and task-based constraints interacted in determining the coordination of hips and ankles during stance. PMID- 15118202 TI - Multiple time scales and multiform dynamics in learning to juggle. AB - To study the acquisition of perceptual-motor skills as an instance of dynamic pattern formation, we examined the evolution of postural sway and eye and head movements in relation to changes in performance, while 13 novices practiced 3 ball cascade juggling for 9 weeks. Ball trajectories, postural sway, and eye and head movements were recorded repeatedly. Performance improved exponentially, both in terms of the number of consecutive throws and the degree of frequency and phase locking between the ball trajectories. These aspects of performance evolved at different time scales, indicating the presence of a temporal hierarchy in learning. Postural sway, and eye and head movements were often 3:2 and sometimes 3:1 frequency locked to the ball trajectories. As a rule, the amplitudes of these oscillatory processes decreased exponentially at rates similar to that of the increase in the degree of phase locking between the balls. In contrast, the coordination between these oscillatory processes evolved exponentially at different time scales, apart from some erratic evolutions. Collectively, these findings indicate that skill acquisition in the perceptual-motor domain involves multiple time scales and multiform dynamics, both in terms of the development of the goal behavior itself and the evolution of the processes subserving this goal behavior. PMID- 15118203 TI - Contextual interference: single task versus multi-task learning. AB - This experiment examined contextual interference in producing a bimanual coordination pattern of 90 degree angle relative phase. Acquisition, retention, and transfer performance were compared in a single-task control group and groups that performed 2 tasks in either a blocked or random presentation. Surprisingly, acquisition data revealed that both the random and control groups outperformed the blocked group. Retention data showed a typical CI effect for performance variability, with the random group outperforming the blocked group. Neither the random nor blocked groups outperformed the control group, suggesting interference of a second task may be as beneficial to learning as extra practice on the initial task. No group effects were found during transfer performance. Results suggest that random practice is beneficial for learning only one task. PMID- 15118204 TI - Investigation of atypical bacteria and virus antigens in respiratory tract infections by use of an immunofluorescence method. AB - In this study an immunofluorescence (IF) method was used to investigate the antigens of viruses and atypical bacteria in respiratory tract infections (RTI) in pediatric and adult age groups. In this prospective study of 2 years (1998 2000), IF was used to investigate the antigens of 7 viral and 3 atypical bacteria to be used for the etiological diagnosis of RTI. Sputum (33.6%) and nasopharyngeal aspirate specimens were obtained from pediatric patients (Group I, 76 cases) and adults (Group II, 135 cases) with RTI symptoms. Antigen detection rates were found to be 44.7% in Group I and 67.4% in Group II (P < 0.05). The following rates for specific antigens in Groups I and II, respectively, were as follows: Chlamydia pneumoniae, 17.1 and 13.3% (P > 0.05); Mycoplasma pneumoniae, 0 and 9.6% (P < 0.05); influenza A virus, 3.9 and 16.3% (P < 0.05); adenovirus, 3.9 and 14.8% (P < 0.05); parainfluenza virus type 1, 5.3 and 7.4% (P > 0.05); respiratory syncytial virus, 9.2 and 1.5% (P < 0.05); parainfluenza virus type 2, 3.9 and 3%(P > 0.05); and influenza B virus, 1.3 and 1.5% (P > 0.05). Mixed agents were found at a rate of 2.6 and 3.7% (P > 0.05) in Groups I and II, respectively. Parainfluenza virus type 3 and Legionella pneumophila antigens were not found. Since detecting etiological agents provides an important guide for determining the most appropriate antibiotic therapy, this IF method could be applied in clinical practice for arriving at a correct diagnosis and administration of effective treatment. PMID- 15118205 TI - An epidemiological analysis of Stenotrophomonas maltophilia strains in a university hospital. AB - The aim of this investigation was to evaluate the epidemiology of Stenotrophomonas maltophilia in a university hospital of Turkey. From June 2000 to December 2001, S. maltophilia strains were collected, clinical presentations were noted, and MIC determinations were performed by means of E-test. Enterobacterial repetitive intergenic consensus sequences-PCR (ERIC-PCR) was used for molecular typing of the strains. Forty-four strains of S. maltophilia were isolated from 41 hospitalized patients in a teaching hospital. The majority of specimens were from the blood and respiratory tract. Antimicrobial sensitivities of these strains were as follows: 97.7 % trimethoprim-sulfamethoxazole, 15.9% ticarcillin, and 95.4% ticarcillin- clavulanate. The strains were evaluated using the ERIC-PCR method. It was of interest to note that epidemiological typing revealed three small outbreaks that were caused by a total of 12 strains. The remaining isolates generated singular DNA patterns. DNA amplification was possible in 38 isolates and yielded 26 different patterns in a period of 20 months, leading to the suggestion that commensal bacteria becomes selected in the presence of a suitable host. PMID- 15118206 TI - Intestinal parasitic infections in HIV/AIDS and HIV seronegative individuals in a teaching hospital, Ethiopia. AB - The magnitude of intestinal parasitic infection in human immunodeficiency virus (HIV) /AIDS patients requires careful consideration in the developing world. However, there have been very few studies addressing this issue in Ethiopia. This study was conducted to determine the prevalence of intestinal parasitic infection in HIV/AIDS patients at Jimma Hospital, Southwest Ethiopia, between January and February 2002. Stool specimens from HIV/AIDS patients and control groups were screened for intestinal parasitic infections using direct and formalin-ether sedimentation concentration methods. Out of 78 HIV/AIDS patients, 52.6% (41/78), and out of 26 HIV-negative individuals, 42.3% (11/26), were infected with one or more types of intestinal protozoa and/or helminthes. The parasites detected among HIV/AIDS patients included Ascaris lumbricoides (30.8%), Blastocystis spp. (14.1%), Entamoeba histolytica (10.3%), Trichuris trichiura (6.4%), Strongyloides stercoralis (5.1%), Giardia lamblia (3.8%), Schistosoma mansoni (2.5%), hookworm species (2.5%), and Taenia spp. (1.3%). Multiple infections were more common among HIV/AIDS patients. Blastocystis spp. were found to be significantly higher in HIV/AIDS patients than in controls (P < 0.05). The magnitude of intestinal parasitic infection was high both in HIV/AIDS patients and in controls. Routine examinations of stool samples for parasites would significantly benefit the HIV infected and uninfected individuals by contributing to reduce morbidity. PMID- 15118207 TI - Infection of cerebrospinal fluid shunts: causative pathogens, clinical features, and outcomes. AB - This retrospective chart review describes the clinical features, pathogens, and outcomes of 46 patients with cerebrospinal fluid (CSF) shunt infections collected over 16 years. The overall CSF shunt infection rate was 2.1%, broken down into 1.7 and 9.3% in adult and pediatric groups, respectively. Fever and progressive consciousness disturbance were the most clinical features in the adult patient group, whereas disturbance of consciousness and abdominal symptoms and signs were the two most common clinical features in the pediatric patient group. The most frequently isolated microorganisms were of the Staphylococcus spp., including Staphylococcus aureus and coagulase negative Staphylococcus, which accounted for 47% of the episodes. Furthermore, increases in polymicrobial and Gram-negative bacilli infections were observed in our study. Due to the high proportion of oxacillin-resistant Staphylococcus spp. and polymicrobial infections, we recommend initial empirical antibiotics with both vancomycin and a third generation cephalosporin for cases in which the causative bacteria has not been identified or for which the results of antimicrobial susceptibility tests are not available. For patients who develop smoldering fevers, progressive disturbed consciousness, seizures, or abdominal fullness after ventriculoperitoneal shunt procedures, CSF shunt infections should be suspected. Although some infections have been managed successfully with antimicrobial therapy alone, the timely use of appropriate antibiotics according to antimicrobial susceptibility testing and the removal of the shunt apparatus are essential for successful treatment. PMID- 15118208 TI - Markers for transfusion-associated hepatitis in north Indian blood donors: prevalence and trends. AB - Transfusion-associated hepatitis is a great problem in developing countries including India due to endemic hepatitis infections and a lack of voluntary donors, trained personnel, and funds. The prevalence of post-transfusion hepatitis B and C in India is about 1-5% and 1%, respectively. A total of 128,589 blood donors were screened for hepatitis B surface antigen (HBsAg) and 76,089 donors were screened for anti-hepatitis C virus (HCV) from 1997 - 2002. Data were tabulated annually. Out of the total 83.6% were replacement donors. Our study concluded that the prevalence of HBsAg and antibodies for HCV ranged between 1.7 2.2% and 0.25 - 0.9%, respectively among all of the donors. Seropositivity was definitely higher in replacement donors than in voluntary donors. Based on these results, we recognize an urgent need to establish a non-remunerated voluntary donor base in India. A stringent deferral system should be developed. The use of sensitive laboratory tests and the addition of core antigen (anti-HBc) to the mandatory screening test list would further reduce the incidence of post transfusion hepatitis. PMID- 15118209 TI - Bactericidal effects of acidic electrolyzed water on the dental unit waterline. AB - Many studies have been conducted in the United States regarding the microbial contamination of dental unit waterline, but not in Japan. Recently, acidic electrolyzed water has been used in the medical and dental fields. In this study, we investigated the bactericidal effects of the temporary inflow of acidic electrolyzed water on microbial contamination of the dental unit waterline. First, in order to observe the daily bacterial contamination of the dental unit waterline, water samples were collected at the end of handpieces and three-way syringes before the inflow of acidic electrolyzed water. They were cultured to detect viable bacteria. Later, the inflow of acidic electrolyzed water was conducted through the piping box of the dental unit. Before starting operation on next day, water samples were collected and cultured, as described above. The mean viable bacteria count was 910 -/+ 190 CFU/ml at the end of handpieces, and 521 /+ 116 CFU/ml at the end of three-way syringes before the inflow of acidic electrolyzed water. However, bacteria were detected in only small numbers at the end of handpieces and three-way syringes on the next day. These results indicated that acidic electrolyzed water could be applied as an appropriate measure against bacterial contamination of the dental unit waterline. PMID- 15118210 TI - Possible horizontal transmission of crimean-congo hemorrhagic Fever virus from a mother to her child. AB - The case of a child with Crimean-Congo hemorrhagic fever (CCHF) presumably infected with CCHF virus from her 27-year-old mother is described. The mother with CCHF was treated with ribavirin and did not present with any symptoms of obvious hemorrhage. The child developed fever on the 5th day after the mother's onset. The partial virus genome was amplified by RT-PCR, and nested PCR from the child and the genome sequence were identical to that from the mother, indicating possible transmission of the virus from mother to child. This case indicates the importance of preventive measures for in-house outbreaks of CCHF. PMID- 15118211 TI - Endotoxin content in Haemophilus influenzae type b vaccine. AB - Haemophilus influenzae type b (Hib) is a major cause of bacterial meningitis among children. Hib conjugate vaccines have effectively prevented Hib infection, and routine immunization with Hib conjugate vaccine has diminished the incidence of the disease in the United States and European countries. Introduction of Hib conjugate vaccines is also required in Japan. However, endotoxin that can carry over from Gram-negative H. influenzae with a purified component may contribute to adverse events following Hib vaccination. In the present study, we examined the endotoxin content in Hib conjugate vaccines. The Hib conjugate vaccine batches, which were produced by a European vaccine manufacturer, were shown to have considerably high endotoxin activity and to vary from 13.9 to 173.7 endotoxin units/dose. These results suggest that it is necessary to monitor the endotoxin content of the vaccine batches to ensure the quality and safety of the vaccines. PMID- 15118212 TI - A comparative survey of serum androgenic hormones levels between male patients with dermatophytosis and normal subjects. AB - Fungal growth can be influenced by human physiological mediators such as androgenic hormones. The aim of this study was to investigate a possible relationship between androgenic hormones and susceptibility to dermatophytosis. To this purpose we measured the levels of testosterone, androstendione, and dehydroepianderosterone sulfate (DHEA-S) in 60 male patients with dermatophytosis due to Epidermophyton floccosum and Trichophyton rubrum by enzyme link immunoassay. Serum testosterone concentration was found to be significantly lower in patients with E. floccosum than in healthy subjects. No significant differences in androstendione and DHEA-S levels were noted between the patients and the healthy individuals. The results showed that testosterone concentration can be considered a predisposing factor for tinea cruris infection. PMID- 15118213 TI - An unexpected outbreak of Japanese encephalitis in the Chugoku district of Japan, 2002. AB - Six patients unexpectedly presented with Japanese encephalitis (JE) from early August to mid-September 2002 in the Chugoku district of Japan. The mean age was 67.5 years (range 42 - 89 years); the onset period in two patients shifted to the middle of September. The JE virus was isolated from the cerebrospinal fluid samples from two patients, and the strain isolated in the one was identified as genotype III. Neurologically, consciousness impairment, meningeal signs, rigidity, hemiparesis, tetraparesis, and convulsive seizures were commonly observed. Magnetic resonance imaging uniformly revealed high signal intensities in the bilateral thalami, brainstem (substantia nigra), hippocampi, and brain cortices. In all patients, acyclovir was used, due to the unexpected outbreak of JE. Five patients, except for one without sequelae, had a severe outcome, including one death. This report indicates that JE in Japan is still a threat to adults and the elderly with decreased or absent immunity to the JE virus. PMID- 15118214 TI - Seroepidemiologic study on pertussis, diphtheria, and tetanus in the Fukuoka area of southern Japan: seroprevalence among persons 0-80 years old and vaccination program. AB - In Japan, mass vaccination for diphtheria, pertussis, and/or tetanus has been mandated by the Vaccination Law since 1948. In order to evaluate the efficacy of this vaccination policy, we conducted seroepidemiological studies on pertussis, diphtheria, and tetanus among individuals aged 0 - 80 years. The pertussis toxin seropositive rates of the vaccine-eligible groups and vaccine-ineligible groups were 55.0 and 57.9%, respectively. The seropositive rate of each group for diphtheria antitoxin was 76.3 and 75.7%, respectively. The tetanus antitoxin seropositive rates were 91.7 and 10.5%, respectively, showing a significant difference between the two groups (P < 0.001). For the three diseases, variations were seen between age groups in the geometric mean antibody titers due to changes of the vaccination program. The results of this study show that natural Bordetella pertussis infection has occurred more frequently than expected. In order to establish the most appropriate vaccination program for the control of pertussis, diphtheria, and tetanus in Japan, further evaluation is necessary. PMID- 15118215 TI - Regional distribution of acute flaccid paralysis cases in Ethiopia in 2000-2002. PMID- 15118216 TI - Prevalence of erythromycin-, tetracycline-, and aminoglycoside- resistance genes in methicillin-resistant Staphylococcus aureus in hospitals in Tokyo and Kumamoto. PMID- 15118217 TI - Molecular epidemiology of Serratia marcescens in a hospital. PMID- 15118218 TI - Clinical pictures of children with human metapneumovirus infection: comparison with respiratory syncytial virus infection. PMID- 15118219 TI - Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a Tokyo hospital in 2003. PMID- 15118220 TI - Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a Kumamoto hospital in 2003. PMID- 15118221 TI - Molecular epidemiology of methicillin-resistant Staphylococcus aureus in a Sendai hospital in 2003. PMID- 15118222 TI - Resetting mechanism of central and peripheral circadian clocks in mammals. AB - Almost all organisms on earth exhibit diurnal rhythms in physiology and behavior under the control of autonomous time-measuring system called circadian clock. The circadian clock is generally reset by environmental time cues, such as light, in order to synchronize with the external 24-h cycles. In mammals, the core oscillator of the circadian clock is composed of transcription/translation-based negative feedback loops regulating the cyclic expression of a limited number of clock genes (such as Per, Cry, Bmal1, etc.) and hundreds of output genes in a well-concerted manner. The central clock controlling the behavioral rhythm is localized in the hypothalamic suprachiasmatic nucleus (SCN), and peripheral clocks are present in other various tissues. The phase of the central clock is amenable to ambient light signal captured by the visual rod-cone photoreceptors and non-visual melanopsin in the retina. These light signals are transmitted to the SCN through the retinohypothalamic tract, and transduced therein by mitogen activated protein kinase and other signaling molecules to induce Per gene expression, which eventually elicits phase-dependent phase shifts of the clock. The central clock controls peripheral clocks directly and indirectly by virtue of neural, humoral, and other signals in a coordinated manner. The change in feeding time resets the peripheral clocks in a SCN-independent manner, possibly by food metabolites and body temperature rhythms. In this article, we will provide an overview of recent molecular and genetic studies on the resetting mechanism of the central and peripheral circadian clocks in mammals. PMID- 15118223 TI - Modulation of a feeding neural circuit by microinjection of K+ channel expression genes into a single identified neuron in Aplysia kurodai. AB - In Aplysia buccal ganglion expression genes for voltage-dependent K(+) channels (AKv1.1a) were injected into one of four electrically coupled multi-action (MA) neurons that directly inhibit jaw-closing (JC) motor neurons and may cooperatively generate their firing pattern during the feeding response. Following the DNA injection, the firing threshold increased and the spike frequency at the same current decreased in the current-induced excitation of the MA neuron; indicating a decrease in excitability of the MA neuron. This procedure also reduced the firing activity of MA neurons during the feeding-like rhythmic responses induced by the electrical nerve stimulation. Moreover, the firing pattern in JC motor neurons was remarkably changed, suggesting the effective contribution of a single MA neuron or electrically coupled MA neurons to the generation of the firing pattern in the JC motor neurons. This method appears useful for exploring the functional roles of specific neurons in complex neural circuits. PMID- 15118224 TI - Intracellular responses of antennal chordotonal sensilla of the American cockroach. AB - The responses of mechanoreceptor neurons in the antennal chordotonal organ have been examined in cockroaches by intracellular recording methods. The chordotonal organ was mechanically stimulated by sinusoidal movement of the flagellum. Stimulus frequencies were varied between 0.5 and 150 Hz. Receptor neurons responded with spike discharges to mechanical stimulation, and were classed into two groups from plots of their average spike frequencies against stimulus frequency. Neurons in one group responded to stimulation over a wide frequency range (from 0.5 to 150 Hz), whereas those in a second group were tuned to higher frequency stimuli. The peak stimulus frequency at which receptor neurons showed maximum responses differed from cell to cell. Some had a peak response at a stimulus frequency given in the present study (from 0.5 to 150 Hz), whereas others were assumed to have peak responses beyond the highest stimulus frequency examined. The timing for the initiation of spikes or of a burst of spikes plotted against each stimulus cycle revealed that spike generation was phase-locked in most cells. Some cells showed phase-independent discharges to stimulation at lower frequency, but increasing stimulus frequencies spike initiation began to assemble at a given phase of the stimulus cycle. The response patterns observed are discussed in relation to the primary process of mechanoreception of the chordotonal organ. PMID- 15118225 TI - Seasonal physiology of the wild raccoon dog (Nyctereutes procyonoides). AB - The raccoon dog (Nyctereutes procyonoides) is a canid omnivore with autumnal fattening and winter sleep. Farmraised raccoon dogs have elevated plasma leptin and growth hormone levels in the winter and depressed plasma cortisol and insulin concentrations during wintertime food deprivation. However, these parameters were not previously tested in the wild population. In the present study 37 wild raccoon dogs were sampled at different seasons and diverse biochemical variables were determined. The results mostly confirmed previous observations on farmraised raccoon dogs. The liver glycogen stores increased during the autumnal fattening period but were low in the winter. The liver glycogen phosphorylase activity decreased but lipase activity increased in the winter indicating the use of fat as the principal metabolic fuel. The plasma insulin concentrations were low in the winter allowing the release of fatty acids from adipose tissue. Low wintertime cortisol and thyroid hormone levels could contribute to protein sparing. Unlike on farms, wild raccoon dogs did not show seasonal fluctuations in their plasma ghrelin or growth hormone levels. The observed physiological phenomena emphasise the adaptation of the species to long periods of food scarcity in the winter. PMID- 15118226 TI - HNF-1 regulates the promoter activity of the HP-27 gene. AB - The hibernation-specific HP-27 gene is expressed specifically in the liver of the chipmunk, a hibernating species of the squirrel family, and exists as a pseudogene in the tree squirrel, a nonhibernating species. In the promoter region, the chipmunk gene has a potential HNF-1 binding site, and the tree squirrel gene has two base substitutions in the corresponding sequence. In this paper, we investigated the role of HNF-1 in the HP-27 gene promoter activity. Gel retardation assays with in vitro-translated HNF-1 and super-shift assays using HepG2 nuclear extracts and an anti-HNF-1 antibody revealed that HNF-1 bound to the chipmunk gene sequence. HNF-1 also bound to the tree squirrel sequence, but with much lower affinity. In HepG2 cells, HNF-1 activated transcription from the chipmunk HP-27 gene, but not from the tree squirrel gene. In addition, the tree squirrel-type base substitutions in the HNF-1 binding site greatly reduced the promoter activity of the chipmunk HP-27 gene. These results indicate that HNF-1 is required for the promoter activity of the chipmunk HP-27 gene, and that the base substitutions in the HNF-1 binding site are involved in the lack of HP-27 gene expression in the tree squirrel. PMID- 15118227 TI - Developmental process of genital ducts in the medaka, Oryzias latipes. AB - The morphological development of genital ducts both intra-gonadal (ovarian cavity and efferent duct) and extra-gonadal (oviduct and sperm duct) was investigated in a model teleost, medaka Oryzias latipes. The results showed that the extra gonadal genital ducts contained two structural units, the anterior and posterior parts, in both sexes. Of special interest is a newly discovered process for the development of a posterior part of the oviduct. The anterior part of oviduct extended continuously from the ovarian cavity at the posterior end of the ovary. Then the posterior part of oviduct, which termed genital pore lip (GPL) in this study, was formed. This part results from invagination and cavitation of the cortex of urinogenital papillae (UGP) and forms the wall of the oviduct opening. We also suggest that the ventral region of urethra mesenchyme has an important role in extra-genital ducts formation. PMID- 15118228 TI - Inhibition of the canonical Wnt signaling pathway in cytoplasm: a novel property of the carboxyl terminal domains of two Xenopus ELL genes. AB - The Wnt signaling pathways are important in many developmental events. The canonical Wnt pathway is one of the three major Wnt-mediated intracellular signaling pathways and is thought to activate Dvl followed by the stabilization of beta-catenin. In Xenopus, this pathway is involved in dorsal determination, anterior-posterior patterning during gastrulation, and neural induction. Here we describe a role for the Xenopus ELL (Eleven-nineteen Lysine-rich Leukemia) gene product in canonical Wnt signaling. Translocation of ELL has been associated with acute myeloid leukemia and the protein possesses three functional domains. We identified rELL-C from a rat brain cDNA library as a binding factor for Dishevelled (Dvl); it represents a partial sequence of rat ELL lacking the pol II elongation domain and has been shown to suppress canonical Wnt signaling. Next, we isolated two Xenopus homologs of ELL, xELL1 and xELL2. No obvious phenotypes were observed with microinjection of full-length xELL1 or xELL2 mRNA, however, microinjection with their occludin homology domain inhibited Wnt signaling at the level of Dvl and upstream of beta-catenin. Intracellular localization of microinjected xELL1- and xELL2-GFP mRNAs showed localization of the full-length products in the nucleus and the occludin-homology domain products in cytoplasm. These results raise the possibility that ELL, which is thought to function as a transcription factor in nuclei, can serve other, novel roles to suppress canonical Wnt signaling in the cytoplasm. PMID- 15118230 TI - Melatonin binding sites in the brain of European sea bass (Dicentrarchus labrax). AB - Characteristics, day-night changes, guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) modulation, and localization of melatonin binding sites in the brain of a marine teleost, European sea bass Dicentrarchus labrax, were studied by radioreceptor assay using 2-[(125)I]iodomelatonin as a radioligand. The specific binding to the sea bass brain membranes was rapid, stable, saturable and reversible. The radioligand binds to a single class of receptor site with the affinity (Kd) of 9.3 +/-0.6 pM and total binding capacity (Bmax) of 39.08 +/-0.86 fmol/mg protein (mean+/-SEM, n=4) at mid-light under light-dark (LD) cycles of 12:12. Day-night changes were observed neither in the Kd nor in the Bmax under LD 12:12. Treatment with GTPgammaS significantly increased the Kd and decreased the Bmax both at mid-light and mid-dark. The binding sites were highly specific for 2 phenylmelatonin, 2-iodomelatonin, melatonin, and 6-chloromelatonin. Distribution of melatonin binding sites in the sea bass brain was uneven: The Bmax was determined to be highest in mesencephalic optic tectum-tegmentum and hypothalamus, intermediate in telencephalon, cerebellum-vestibulolateral lobe and medulla oblongata-spinal cord, and lowest in olfactory bulbs with the Kd in the low picomolar range. These results indicate that melatonin released from the pineal organ and/or retina plays neuromodulatory roles in the sea bass brain via G protein-coupled melatonin receptors. PMID- 15118229 TI - Characterization of two types of cytochrome P450 aromatase in the serial-sex changing gobiid fish, Trimma okinawae. AB - To investigate the role of estrogen in the serial-sex changing fish Trimma okinawae, we isolated complementary DNAs encoding two distinct cytochrome P450 aromatase isoforms from adult ovary and brain (termed P450aromA and P450aromB, respectively). Sequence and phylogenic analyses showed that the goby P450arom forms belong to two separate CYP19 subfamilies. Transient expression of these cDNAs in HEK293 cells caused conversion of exogenous testosterone to estradiol 17beta. RT-PCR showed that P450aromA was expressed in the brain, spleen, testis and ovary. P450aromB was expressed in the brain, liver, testis and ovary. In situ hybridization studies showed that P450aromA mRNA, but not P450aromB mRNA, was present in both ovary and testis. Positive signals were restricted to granulosa cells of vitellogenic follicles and interstitial cells of mature testis. Ovarian expression of both P450arom genes during the spawning cycle was examined by quantitative real-time RT-PCR. P450aromA transcripts increased during vitellogenesis and decreased prior to spawning. In contrast, P450aromB transcripts were barely detectable and did not correlate with ovarian development. These findings suggest that P450aromA, but not P450aromB, is involved in regulating ovarian vitellogenesis in goby. PMID- 15118231 TI - Development of gonadotropes in the chicken embryonic pituitary gland. AB - Although a number of immunohistochemical studies have been carried out on the differentiation of chicken gonadotropes during embryogenesis, the temporal and spatial properties of appearance of gonadotropes are not clear. In this study, we studied the appearance and morphological characteristics of gonadotropes in the embryonic and adult chicken anterior pituitary glands using RT-PCR, in situ hybridization and immunohistochemistry. For this purpose, we raised specific antisera against chicken follicle-stimulating hormone beta-subunit (cFSHbeta) and chicken luteinizing hormone beta-subunit (cLHbeta) based on each putative amino acid sequence. RT-PCR analysis revealed that cFSHbeta mRNA was expressed from embryonic day 7 (E7). Chicken FSHbeta mRNA-expressing (-ex) and -immunopositive ( ip) cells started to appear in the ventral part of the caudal lobe in the anterior pituitary gland at E8. Chicken LHbeta-ip cells were also first observed there at E8, but cLH mRNA expression was confirmed from E4 by RT-PCR analysis. The distribution of these chicken gonadotropin-ex and -ip cells spread from the ventral part to dorsal part in the caudal lobe around E10 and subsequently expanded to the cephalic lobe from E12 to E20. These cells were morphologically classified into two types (round- and club-shaped cells). It was found that the density of gonadotropin-ip cells in the caudal lobe was always higher than that in the cephalic lobe throughout the period of development. To the best of our knowledge, this is the first report focusing on the differentiation of chicken gonadotropes by assessment of both protein and mRNA of chicken gonadotropin. PMID- 15118232 TI - Comparative morphology of the stolonic vessel in a didemnid ascidian and some related tissues in colonial ascidians. AB - The stolonic vessel is a tubular projection of the epidermis from the anterior part of the abdomen in the didemnid ascidians, and the vessel has been supposed to be closely related to the stolons, vascular appendages, and the posterior ends of the abdomen in other aplousobranch ascidians. We compared the morphology of the stolonic vessels of Diplosoma virens with similar or related tissue in other colonial ascidians, e.g. stolons of Clavelina, vascular appendages of Distaplia and Eudistoma, tunic vesicle of Aplidium, and vascular ampullae of Botrylloides. The epidermis of the stolonic vessel is composed of cuboidal cells in lateral wall and columnar cells at the distal tip of the vessel. The cuboidal cells have microvilli that probably anchor the stolonic vessel to the tunic. The columnar cells contain round granules that may concern with the secretion of some tunic components. The secretion of the granules, however, could not observed in this study. The stolonic vessel of D. virens is similar in morphology to the vascular ampullae of Botrylloides and the tunic vesicle of Aplidium rather than the other tissue examined here. Since the cell morphology is supposed to reflect its function but not the phylogenetic relationship, the present study could not provide conclusive evidences to prove the homology and the phylogenetic relationship among the tubular, epidermal projections in the colonial ascidians. PMID- 15118233 TI - Two new species of Spiochaetopterus (Annelida: Polychaeta) from Sagami Bay and Tokyo Bay, Central Japan with a comparative table of species from Japanese and adjacent waters. AB - Two new species of Spiochaetopterus (Chaetopteridae: Polychaeta), S. sanbanzensis, from Sanbanze, off Ichikawa and Funabashi Cities, Tokyo Bay, and S. izuensis from the shallow waters of Sagami Bay, were described. The most obvious difference is the number of segments in region A: 9 in S. sanbanzensis and 10 in S. izuensis. In addition, Spiochaetopterus sanbanzensis has elliptical light brown or blackish eye-spots, asymmetrical cordate specialized A4 chaetae, and a color pattern consisting of many dispersed brown spots on both ventral and dorsal faces of region A. In Spiochaetopterus izuensis, a brown band extends from each eye-spot to the level of the A1 chaetae; the convex ventral edge of the head of the specialized A4 chaeta has an oblique section and the color pattern of the body is absent. A comparison is established between these two new species and other known species from Indo-Pacific Ocean. PMID- 15118234 TI - A new species of the subterranean genus Krenedrilus dumnicka (Oligochaeta, Tubificidae) from the deep bottom of an oligotrophic caldera lake in Japan. AB - A new tubificid oligochaete, Krenedrilus towadensis, is described from deep bottoms of the oligotrophic caldera Lake Towada in northern Japan. It resembles K. realis Martinez-Ansemil and Collado, 1996, in the combination of somatic setae, but it is different from all other congeners by the lack of spermathecal setae and epidermal papillae in X, in the structures of penial and supernumerary setae, and in the location of spermathecal pores. The definition of Krenedrilus is revised. PMID- 15118235 TI - Bottleneck effects on the sika deer Cervus nippon population in Hokkaido, revealed by ancient DNA analysis. AB - The population size of the sika deer Cervus nippon on Hokkaido Island of Japan had been remarkably reduced because of heavy hunting pressure since the beginning of Meiji Period and effects of heavy snow in 1879 and 1881. After that, the number of sika deer in Hokkaido has increased gradually due to the protection by the Hokkaido government. In the present study, in order to investigate the bottleneck effects, we analyzed ancient mitochondrial DNA (mtDNA) on sika deer bones excavated from archaeological sites just before Meiji Period. On 86 of 113 bones from 13 archaeological sites of Ainu Culture Period (17-19th centuries), 602 base-pair fragments of the mtDNA control region were successfully sequenced. Consequently, we found three new haplotypes (g-, h- and i-types) which had not been identified in modern sika deer. In addition, four haplotypes (a-, b-, c- and d-types) identified from modern sika deer were also found in the archaeological deer. The new haplotypes and previously reported hapoltypes from sika deer of Hokkaido were phylogenetically much closer to each other, compared with those of modern sika deer from Honshu, Kyushu and the Chinese continent. Geographical distribution patterns of haplotypes of the ancient population were different from those of the modern population in Hokkaido. Our findings indicated that their genetic diversity was reduced through the bottleneck and that population structures of sika deer were changed widely in Hokkaido due to genetic drift. PMID- 15118236 TI - Geographic variation and diversity of the cytochrome b gene in wild populations of medaka (Oryzias latipes) from Korea and China. AB - We analyzed the polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLPs) of the mitochondrial cytochrome b (cyt b) gene in wild populations of medaka from Korea and China. We surveyed 258 wild specimens from 75 different sites, and identified 17 mitotypes. Sequencing analysis of the complete cyt b gene (1141-bp) was subsequently carried out to infer the phylogenetic relationships among these mitotypes. Phylogenetic trees indicated two major clades, D and E, which were different from the Japanese clades (A, B and C). These two clades were completely identical to two clusters previously identified by RFLP analysis of entire mitochondrial DNAs. The geographic distribution of the mitotypes in clades D and E was consistent with the China West Korean Population and the East Korean Population as defined by allozymic and karyological analyses. This agreement among different analyses suggests long-term isolation between the two groups. In the region where the distributions of two major clades overlapped, a limited extent of gene flow was observed. These results suggested the existence of some reproductive isolation mechanisms between the two clades, or the introgression between them followed by a random drift in each local population. Furthermore, clade D was subdivided into three subclades (D-I to D-III). The phylogenetic relationship and distribution pattern of subclade D-II suggested a dispersal event of medaka from China to southwest Korea. Our results also showed that the East Korean Population has recently expanded its distribution area because little diversity was observed in clade E. PMID- 15118237 TI - Proton pumping ATPases and diverse inside-acidic compartments. AB - Proton-translocating ATPases are essential cellular energy converters that transduce the chemical energy of ATP hydrolysis into transmembrane proton electrochemical potential differences. The structures, catalytic mechanism, and cellular functions of three major classes of ATPases including the F-type, V type, and P-type ATPase are discussed in this review. Physiological roles of the acidic organelles and compartments contained are also discussed. PMID- 15118238 TI - Development of cell model with specific functions and its application to the study of global gene expression. AB - The use of in vitro cell culture models has been of central importance in the development of our understanding of the cellular and molecular biology of organs and tissues. Transgenic mice and rats harboring temperature-sensitive simian virus 40 large T-antigen genes are useful for establishing cell lines from organs and tissues that have proved difficult to culture in vitro. Many conditionally immortalized cell lines with normal specific functions were generated from transgenic animals. DNA microarray technology has broad applications and is directed toward the study of global gene expression. Using established cell lines and DNA microarrays, we identified many genes that were up- and down-regulated in the process of the cell differentiation or cell death. In this review, the characteristics of established cell lines and possible applications of the study of global gene expression are discussed. PMID- 15118239 TI - Structure-activity relationship studies on cyclic RGD peptides utilizing novel alkene dipeptide isosteres. AB - A structure-activity relationship study was performed on cyclic RGD peptides using a combination of multisubstituted alkene dipeptide isosteres. To clarify the effects on bioactivity of a valine N-methyl group in the cyclo(-Arg-Gly-Asp-D Phe-MeVal-) peptide developed by Kessler's group, novel D-Phe-Val-type isosteres with methyl-substituting groups on the olefin were designed and synthesized. Syntheses of D-Phe-psi[(E)-CH=CMe]-Val-type isosteres were carried out in essentially identical fashion to the previously reported preparation of psi[(E) CH=CH]-type congeners. Alternatively, D-Phe-psi[(E)-CMe=CX]-Val-type isosteres (X=H or Me) were synthesized via stereoselective alkylation of beta-(1,3 oxazolidin-2-on-5-yl)-alpha,beta-enoates using organocopper reagents. The resulting four isosteres were utilized in either solution- or solid-phase peptide synthesis to afford the cyclic RGD peptidomimetics, cyclo(-Arg-Gly-Asp-D-Phe psi[(E)-CX=CX]-Val-) (X=H or Me). alpha(V)beta(3) and alpha(IIb)beta(3) integrin antagonistic activities of the peptidomimetics along with Kessler's peptides were comparatively evaluated. In addition, structural calculations of these compounds by simulated annealing/energy minimization using dihedral and distance restraints derived from (1)H-NMR data in DMSO gave insight into the effects of the valine N methyl group as well as the D-phenylalanine carbonyl oxygen. PMID- 15118240 TI - Development and evaluation of pharmaceutical services in the ICU/CCU by medical staffs. AB - Questionnaires were sent out to the staffs (13 physicians, 52 nurses and 5 medical engineers) of the ICU/CCU at the University of Tokyo Hospital, to evaluate pharmaceutical services by analyzing problems in the services offered. Four components of pharmaceutical services were evaluated: inventory control of drugs, check of drug usage and doses, mixing of injections, and offering drug information. Almost all responses from medical staffs evaluated pharmaceutical services overall as "good". The high response rate (96%) from the nursing staff was attributed to the fact that they were familiar with the pharmacist's role with drug inventory, and mixing injections, when nursing was not available for these tasks. Although 50% of physicians rated the pharmaceutical services of providing drug information as "good", this value was lower than responses on other items of the questionnaires, which suggests some dissatisfaction. The occurrences of drug information obtained by passive offering (121 subjects) was 4 times as common as drug information obtained by active offering (30 subjects). From this finding, and comments on the questionnaires from physicians, it suggests that physicians require more drug information for dosage regimens, and prefer the drug information to be provided more actively. Further, an important comment from physicians and nurses was that the services of pharmacists are not available on all shifts/all days of the week to provide consultation for drug information and mixing of injections. Although having a pharmacist available daily around the clock is desirable and ideal to the medical team, the number of pharmacists under the present system cannot support this. As a solution, we think that it is crucial that pharmacists educate medical staff when they are present to in order to optimize therapy and patient care over time. PMID- 15118241 TI - A study of transdermal fentanyl in cancer pain at Aichi-Cancer Center. AB - In Japan, transdermal fentanyl (Durotep Patch) was launched in March 2002, and it was regarded as making opioid rotation possible. When changing from morphine to transdermal fentanyl, the efficacy ratio of 1:150 is used in Japan as well as in many other countries. However, the ratio of 1:100 is used in Germany. As a result, a dose increase in transdermal fentanyl is often required to control pain. We studied transdermal fentanyl use in the Aichi Cancer Center (ACC) to investigate the actual conversion ratio and appropriate switching by following up 144 patients (81 men, 63 women) who had received transdermal fentanyl in the ACC from March 19, 2002, to April 30, 2003. Transdermal fentanyl improved pain control in patients who had difficulty in tolerating oral medication or in continuing morphine because of side effects. Regression analysis indicated that the efficacy ratio of oral morphine to transdermal fentanyl was 1:78. As the fentanyl dosage was excessive even in some patients who followed the recommended morphine/fentanyl conversion of 150:1, it is dangerous to use the conversion ratio of 78:1 at first. Morphine side effects were reduced in some patients who changed to transdermal fentanyl, but there was no reduction in those who needed high-dose morphine for rescue analgesia. Therefore it is safe and effective to use low-dose transdermal fentanyl in the beginning and to control pain promptly using rescue morphine based on the present recommended dosage. For opioid rotation, quick-acting opioids other than morphine are expected to be launched in Japan. PMID- 15118242 TI - Pre-implantation conceptus and maternal uterine communications: molecular events leading to successful implantation. AB - Implantation, a critical step for mammals in establishing pregnancy, requires successful completion of sequential events such as maternal uterine development, conceptus development and attachment, and placental formation. To reach the stage of placental formation, synchronized development of the conceptus and uterus throughout the implantation period is absolutely required. A number of factors expressed at the uterine endometrium and/or conceptus, which are associated with peri-implantation development, have been identified. In addition to a temporal and spatial expression of these factors, their roles in intra- and inter-cellular interactions make it difficult to fully understand physiological roles played during the critical period. This paper focuses on early conceptus development, maternal preparation for implantation and uterine-conceptus communication during the pre-implantation period, rather than the subsequent events such as conceptus attachment to the maternal endometrium. New aspects of pre-implantation processes are evaluated through simultaneous expressions of transcription factors as they possibly regulate the complex processes of implantation events in murine species and ruminant ungulates. PMID- 15118243 TI - Activation with ethanol improves embryo development of ICSI-derived oocytes by regulation of kinetics of MPF activity. AB - Developmental potential of bovine embryos that are not artificially activated after intracytoplasmic sperm injection (ICSI) is generally very low. In this study, we investigated effects of artificial activation with ethanol on kinetics of maturation promoting factor (MPF) activity (p34(cdc2) kinase activity) and development of bovine oocytes following ICSI. Treatment of oocytes with ethanol at 4 h after ICSI improved their first cleavage and further preimplantation development (51% vs. 13%, 14% vs. 4%: treatment with vs. without ethanol, respectively). MPF activity of oocytes was lowered until at least 2 h after ICSI. In oocytes without activation after ICSI, MPF activity temporarily elevated at 6 h after ICSI, whereas this phenomena was not observed in the oocytes treated with ethanol. Furthermore, MPF activity was elevated 20 h after ICSI in oocytes activated with ethanol, whereas this elevation of MPF activity was not shown in oocytes without activation. These results indicate that the stimulus of sperm was sufficient to lower MPF activity of oocytes following ICSI, and moreover the activation treatment of bovine oocytes with ethanol after ICSI served to maintain the low levels of MPF activity until the next cell cycle started. PMID- 15118244 TI - A phenotypic study of murine oocyte death in vivo. AB - Most studies of oocyte apoptosis have been performed in vitro and have employed the method of artificial induction of apoptosis by an anti-cancer agent. However, the process of oocyte death in vivo has not been clearly identified. To investigate the death process in unfertilized oocytes in vivo, we examined the cytochemical change of oocytes collected by oviduct flushing at various intervals after hCG injection. At each collection time, the collected oocytes were phenotypically classified under the microscope into four groups: single-cell oocytes (non-activated and without a nucleus and cytokinesis), activated oocytes (single-, 2- or 4-cell with a nucleus), fragmented oocytes, and dead oocytes. The number of single oocytes decreased and dead oocytes increased with the lapse of time, but the number of activated oocytes or fragmented oocytes did not. Also, most of the dead oocytes observed were single cell. At each time point, single oocytes were stained with anti-tubulin antibody to examine their spindle status. At 24 h after hCG injection, all ovulated oocytes had a normal bipolar spindle, while at 64 h all single-cell oocytes had no spindle. From these observations, we concluded that most oocyte deaths in vivo occur in the single oocyte stage, not in activated or fragmented oocytes. PMID- 15118245 TI - Anesthesia and acoustic stress-induced intra-uterine growth retardation in mice. AB - Stress interferes with reproduction, adversely influencing implantation and fetal growth, and sometimes even leading to abortion. Here, we attempted to evaluate the early gestational effects of uncomfortable sound on pregnant mice and their offspring. Ten-week-old pregnant Jcl:ICR mice were exposed to sound (100 dB, random frequency between 9-34 kHz) for 8 hours on the 3(rd), 5(th) and 7(th) gestational days (GD). The effects of general anesthesia were also investigated, with or without acoustic stress. All groups were examined on the 18(th) GD for fetal growth. Fetal weight, number of ossified sacrococcygeal vertebrae and placental weight were all significantly reduced (P<0.0001) when stress was induced on the 7(th) GD, but not on the 3(rd) or 5(th) GD. This intra-uterine growth retardation (IUGR) was significantly inhibited by general anesthesia (P<0.0001), although general anesthesia alone induced significant IUGR (P<0.0001) when compared with control mice. This suggests that acoustic exposure indirectly exerts an effect on fetal growth, possibly via a psycho-maternal pathway. We also found that analysis of the number of ossified sacrococcygeal vertebrae is the most sensitive tool for the study of IUGR. PMID- 15118246 TI - Donor and recipient rat strains affect full-term development of one-cell zygotes cultured to morulae/blastocysts. AB - The present study was conducted to examine the developmental potential to offspring of rat embryos cultured from 1-cell to morula/blastocyst stage. Pronuclear zygotes from Wistar x Wistar or (SD x DA) x Wistar strains were cultured in modified rat 1-cell embryo culture medium (mR1ECM) for 96 h in 5% CO(2) in air at 37 C. The proportion of the 3-way cross hybrid zygotes developing into morula/blastocyst stage (74%) was higher than that of the Wistar zygotes (66%). Day-5 morulae/blastocysts developed in vitro were transferred into Day-3 or -4 pseudopregnant recipients of Wistar or SD x DA strain. The transfer of cultured embryos resulted in the birth of offspring at 13-59%, while that of non cultured control blastocysts showed birth rates of 35-65%. The best offspring rate of cultured embryos (59%) was obtained when the hybrid 1-cell zygotes were cultured in mR1ECM medium and transferred into the 2-days earlier uteri of SD x DA recipients. These results suggest that genetic background of recipients as well as donors is a possible factor affecting full-term development of rat morulae/blastocysts derived from 1-cell stage zygotes cultured in vitro. PMID- 15118247 TI - Body growth and plasma concentrations of metabolites and metabolic hormones during the pubertal period in female Shiba goats. AB - It has been shown in various species that the onset of puberty is closely associated with body growth and nutritional state rather than age. The present study was conducted to determine the timing of puberty and to clarify body growth and metabolic changes around the pubertal period in female Shiba goats. Blood samples were collected between 10 to 38 weeks of age from 12 female goats, and plasma concentrations of progesterone, metabolites (glucose, nonesterified fatty acid, ketone body and acetic acid) and metabolic hormones (insulin and insulin like growth factor-I (IGF-I)) were analyzed. Physical parameters (body weight, withers height and body length) were also measured at the blood sampling. The week when plasma progesterone concentrations first exceeded 1.0 ng/ml was designated as the onset of puberty. The results showed that the average age of the onset of puberty was 27.0 +/- 0.9 (mean +/- SEM) weeks in female Shiba goats. When the goats reached puberty, the average values of body weight and goat body mass index ((body weight (kg)/withers height (cm)/body length (cm)) x 10(3)) were 12.2 +/- 0.5 kg and 5.7 +/- 0.2, respectively. No particular change associated with puberty was apparent for plasma concentrations of the metabolites examined. Plasma insulin concentrations were maintained at lower levels until the onset of puberty, and then they began to gradually increase. Plasma IGF-I concentrations began to gradually increase 1 to 4 weeks before the onset of puberty and this increase continued throughout the peripubertal period. These results imply that IGF-I acts as a peripheral nutritional signal to trigger the onset of puberty in Shiba goats. PMID- 15118248 TI - Transgenic expression of testis-specific poly(A) polymerase TPAP in wild-type and TPAP-deficient mice. AB - We have previously identified a testis-specific poly(A) polymerase, TPAP (PAPbeta), involved in poly(A) tail extension of specific mRNAs in the cytoplasm of round spermatids. Targeted disruption of the mouse TPAP gene resulted in the arrest of spermiogenesis due to reduced expression of haploid-specific genes required for morphogenesis of germ cells. To further elucidate the role(s) of TPAP in spermatogenesis, transgenic mice expressing an exogenous TPAP transgene on wild-type and TPAP-deficient backgrounds were generated and characterized. The transgenic mice overexpressing TPAP exhibited normal spermatogenesis and fertility. The sizes of some transcription factor mRNAs as the substrates of TPAP were also unaffected. Transgenic expression of the TPAP gene in the TPAP deficient mice complemented both the incomplete elongation of the poly(A) tails of specific transcription factor mRNAs, and reduced expression of haploid specific genes, resulting in the resumption of normal spermiogenesis. These data conclusively show that spermatogenesis requires the cytoplasmic elongation of the mRNA poly(A) tails catalyzed by TPAP, and imply the presence of a regulatory mechanism(s) defining the extent of the cytoplasmic mRNA polyadenylation. PMID- 15118249 TI - Withdrawal of ovarian steroids stimulates prostaglandin F2alpha production through nuclear factor-kappaB activation via oxygen radicals in human endometrial stromal cells: potential relevance to menstruation. AB - The present study was undertaken to investigate whether withdrawal of estrogen and progesterone (EP-withdrawal) stimulates prostaglandin F2alpha (PGF2alpha) production through oxygen radical (ROS)-induced NF-kappaB activation in human endometrial stromal cells (ESC). To study the EP-withdrawal, ESC that had been treated with estradiol (E, 10(-8) M) and medroxyprogesterone acetate (MPA, 10(-6) M) for 12 days were then incubated with or without E+MPA for a further 11 days. PGF2alpha concentrations in the medium and cyclooxygenase-2 (COX-2) mRNA levels were significantly increased after EP-withdrawal, while they were unchanged by the continuous treatment with E+MPA. When ESC were incubated with N-acetyl-L cysteine (Nac, 50 mM), an antioxidant, during EP-withdrawal, Nac blocked the increases in PGF2alpha production and COX-2 mRNA expression caused by EP withdrawal. Next, we examined whether ROS generated in response to EP-withdrawal acted through NF-kappaB activation. Electrophoretic mobility shift assay revealed that EP-withdrawal caused marked increases in NF-kappaB DNA binding activity, which was completely suppressed by Nac. Furthermore, when ESC were incubated with MG132 (3 microM), which inhibits NF-kappaB activation, during EP-withdrawal, MG132 blocked the increases in PGF2alpha production and COX-2 mRNA expression caused by EP-withdrawal. In conclusion, EP-withdrawal stimulates COX-2 expression and PGF2alpha production through ROS-induced NF-kappaB activation, suggesting a possible mechanism for menstruation. PMID- 15118250 TI - Effects of trichostatin a, a histone deacetylase inhibitor, on mouse gonadal development in vitro. AB - Sry, Sox9 and M33 are thought to act as architectural transcription factors or as a chromatin regulator in gonadal development. However, the direct relationship between chromatin structure and sex determination has not yet been revealed. To clarify the effect of chromatin structural change on gonadal development, we examined the effects of trichostatin A, a histone deacetylase inhibitor, on mouse gonadal development in vitro. In the 0.1 microM treated testicular explants, the size of the gonad was significantly decreased, although the testicular cord formation occurred normally. In the 1.0 microM treated explants, the gonads revealed one or two large testicular cords. Sox9 and MIS expressions suggest that Sertoli cell differentiation is induced normally within the testicular cord, while Dnmt3b expression suggests that several immature Sertoli cells are located on the outside of the testicular cord. The 3beta-hsd expression indicates that Leydig cell differentiation occurs normally. On the other hand, germ cell loss was observed in the treated testicular explants. In the treated ovarian explants, the number of premeiotic germ cells was reduced without gonadal size change. Thus, trichostatin A affects the development of germ cells, but does not affect sex determination. PMID- 15118251 TI - Improved isolation and culture of embryonic stem cells from Chinese miniature pig. AB - Pigs serve as a better research model for human beings than other species. The Chinese laboratory miniature pig is a new laboratory animal and is expected to be applicable in many medical research fields. This study was to establish effective technologies to isolate and culture ES cells in Chinese miniature pigs. For isolation of the inner cell mass from blastocysts, an enzyme-digestive method was compared with the traditional immunosurgery. Isolated ICM were cultured in three feeder cell layers: mouse embryonic fibroblasts (MEF), porcine embryonic fibroblasts (PEF) and a continuous cell line of mouse embryonic fibroblasts (STO). Microtubule activity of the three feeder cells was further examined by immunofluorescence. ICM were successfully isolated from 85% of blastocysts by the enzyme-digestive method, compared to only 40% by immunosurgery. When ICM were cultured in three feeder layers for two to three days, 75%, 65% and 20% of ICMs formed primary cell colonies in MEF, PEF and STO, respectively. Colonies were also formed during subcultures after 9, 5 and 1 passage in MEF, PEF and STO, respectively. Microtubules in STO cells were significantly fewer than those in MEF and PEF. When the ES-like cells were cultured in a differentiation medium, they differentiated to neuron-like cells and other types of cells. These results indicate that healthier ICM can be obtained with the enzyme-digestive method. Successful culture of ICM to ES-like cells has been achieved not only in MEF, but also in homologous (pig) feeder layer. The ES cells obtained in the present study were pluripotent. PMID- 15118252 TI - Assessing estrogenic activity of pyrethroid insecticides using in vitro combination assays. AB - Pyrethroid insecticides are among the most commonly used classes of insecticides worldwide, but their endocrine disrupting activities remain unclear. Therefore, in the present study, we examined the estrogenic activities of pyrethroid insecticides in E-screen and competition binding assays. In addition, we measured estrogen receptor (ER) protein and pS2 mRNA levels in human breast cancer cells (MCF-7 BUS) to clarify the mechanism of their estrogenicity. Seven pyrethroid insecticides (bioallethrine, cypermethrin, deltamethrin, fenvalerate, permethrin, sumithrin, and tetramethrin) were tested because of their worldwide usage. In addition, 17beta-estradiol was tested as a positive control. As expected, 17beta estradiol significantly increased MCF-7 BUS cell proliferation at concentrations of 10(-11) M and above. Of the pyrethroid insecticides tested, only sumithrin increased MCF-7 BUS cell proliferation in a dose-dependent manner; the maximum induction of cell proliferation was observed at a dose of 10(-5) M. In the anti estrogenic activity test, bioallethrin, fenvalerate, and permethrin significantly inhibited 17beta-estradiol-induced MCF-7 BUS cell proliferation at 10(-6) M, a concentration comparable to the effective dose (10(-9) M) of ICI 182,780, a pure ER antagonist. However, none of the pyrethroid insecticides competitively inhibited the binding of [(3)H]estradiol to rat uterus ERs in competition binding assays. Both 17beta-estradiol (10(-10) M) and sumithrin (10(-5) M) decreased the levels of cytosolic ERalpha and ERbeta protein expression significantly as compared with the vehicle control. In addition, 17beta-estradiol (10(-10) M) increased pS2 mRNA expression markedly, and sumithrin significantly increased pS2 mRNA levels in a dose-dependent manner. The other six compounds tested in the present study did not affect ER protein levels or pS2 mRNA levels. These results suggest that certain pyrethroid insecticides may be considered to be estrogen like chemicals that act through pathways other than direct ER binding, and may function as endocrine modulators in both wildlife and humans. PMID- 15118253 TI - Progesterone stimulation by LH involves the phospholipase-C pathway in bovine luteal cells. AB - Luteinizing hormone (LH)-stimulated steroidogenesis in luteal cells is known to be mediated through the activation of cyclic AMP (cAMP)-dependent protein kinase, and to be also modulated by calcium-dependent mechanisms. In the present study, we tested the hypothesis that LH stimulates progesterone (P4) production in bovine luteal cells through activation of phospholipase (PL) C by using a cell culture system. Bovine mid-luteal cells (Days 8-12 of the estrous cycle) were cultured for 24 h and then exposed to a PLC inhibitor (U-73122; 10 microM) with or without LH (10 ng/ml) for 4 h. U-73122 blocked LH-stimulated P4 production without affecting cAMP accumulation. Moreover, exposure of luteal cells to PLC increased P4 production in a dose-dependent manner. These results support the hypothesis that the luteotropic action of LH in bovine luteal cells is mediated not only by activation of adenylate cyclase but also by activation of PLC. PMID- 15118254 TI - Effects of nuclear transfer procedures on ES cell cloning efficiency in the mouse. AB - Enucleated oocytes receiving mouse embryonic stem (ES) cells develop into fertile young. The developmental potential to young is low, however, and the rate of postnatal death is high. We examined the effect of various nuclear transfer procedures on the in vitro and in vivo developmental potential of nuclear transferred oocytes. The potential of oocytes receiving ES cells at M phase to develop into blastocysts after fusion by Sendai virus was high compared with that after direct injection (67% vs. 30%). The developmental potential of oocytes receiving ES cells at the M phase is higher than that of oocytes receiving ES cells at the G(1) phase (30-67% vs. 2-5%). Developmental ability to live young was low in all groups (0-4%). Different activation protocols affected the potential to develop into blastocysts to a different extent (27-62%), but did not affect the potential to develop into live young (0-3%). The present study demonstrated that the various conditions examined did not affect the potential of nuclear-transferred oocytes receiving ES cells to develop into live young or the incidence of postnatal death. PMID- 15118255 TI - [Taurine is a possible anti-atherosclerotic agent]. AB - Atherosclerosis-related ischemic heart diseases are the principal cause of death in the last few years. Recently, several reports implicated that taurine, sulfur containing beta-amino acid, prevented the progression of atherosclerosis through various anti-pathogenetic modifications. Firstly, taurine treatment inhibited lipid peroxidation and/or lowered serum LDL/VLDL cholesterol and elevated HDL, and as a result, it prevented lipid accumulation on the aortic valve in hypercholesterolaemic animals. Secondly, taurine administration prevented endothelial dysfunction, one of the initial events in the formation of lesions of atherosclerosis, through the amelioration of the impairment of monocyte function. Thirdly, while it is well known that taurine scavenges hypochlorous acid (HOCl) produced by myeloperoxidase in neutrophils and macrophages, recent studies revealed that HOCl was one of the major factors oxidizing LDL, implying that the anti-oxidative role of taurine contributes to the anti-atherosclerotic effect. Additionally, TauCl, produced by the reaction of taurine with HOCl, inhibits the activation of NF-kappaB followed by the inhibition of the production of the pro inflammatory mediators. PMID- 15118256 TI - [Mechanisms of psychotropics' action in relation to CNS neurogenesis]. AB - Newly available psychotropics seem to put the practical psychiatry to a stage of reform. In addition, the recent advancements in the study of neurogenesis in the adult brain force the change of the therapeutic strategy of mental disorders. The fact that the central nervous tissues can repair even after the maturation and that the replacement of neurons continues during adulthood will alter our understanding about their pathogenesis. The action of several psychiatric medications such as antidepressants, mood stabilizers, antipsychotics, and electroconvulsive therapy is converging at neurogenesis and/or neuroprotection. When the validity of the "neurogenesis/neuroprotection hypothesis" of psychiatric medication will be proved in psychiatric practice, we can establish a more rational and more effective treatment of mental disorders. PMID- 15118257 TI - [The research method for investigating the role of the mitochondrial permeability transition pore in cell death]. AB - Mitochondria play a key role in the regulation of cell death, necrosis, and apoptosis. It is well known that the opening of the mitochondrial permeability transition pore (PTP) induces the cytochrome C release and results in apoptosis. Also, it has been reported that the inhibition of PTP by cyclosporin A prevents the ischemia/reperfusion-induced necrosis. Thus, the opening of mitochondrial PTP may be a central coordinating event of apoptotic and necrotic cell death. However, the relationship between PTP opening and both modes of cell death remains elusive. In this paper, we report the recent state of our research method and the role of mitochondrial PTP in cardiac cell death. PMID- 15118258 TI - [Recent development of new drugs for the treatment of allergic diseases]. AB - Due to the prevalence of allergic diseases such as bronchial asthma, allergic rhinoconjunctivitis and dermallergosis, efforts at the discovery of novel and effective medications for prevention and treatment of these conditions have been reinforced. Recently, it has been recognized that these allergic diseases are a chronic inflammatory disorder of the lower and upper airways and skin. In this article, we reviewed the recent development of the following new antiallergic therapies: anti-Th2 cytokine antibodies, decoy receptors, receptor antibodies, anti-IgE antibodies, anti-cell adhesion molecules antibodies, antisense oligonucleotides, keratinocyte modulators, inhibitors of phosphodiesterase 4, tachykinin receptor antagonists, and anti-histaminic drugs. Most of these new agents are aimed to inhibit various components of allergic inflammation. The future use of allergic disease therapies hold great promise and excitement. PMID- 15118259 TI - [Pharmacological and pharmacokinetic features and clinical effects of pitavastatin (Livalo Tablet)]. AB - Today 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors (statins) are the most often prescribed drugs among the therapeutics for hypercholesterolemia. Pitavastatin is a novel statin that has been developed entirely in Japan from the biological screening to clinical studies persuing more efficatious statin than hitherto known. Preclinical studies on drug metabolism revealed that pitavastatin is distributed selectively to the liver, excreted into bile without metabolic modification, and efficiently re-circulates to the liver to show a prolonged plasma half-life. In guinea pigs, pitavastatin enhanced hepatic LDL receptor activity and reduced VLDL secretion in a liver perfusion study, and it lowered plasma total cholesterol (TC) levels at 0.3 mg/kg and triglyceride (TG) levels at 1 mg/kg, respectively, and more. From these results, pitavastatin is assumed to lower LDL cholesterol (LDL-C) by promoting LDL receptor expression and further potentiate the cholesterol-lowering effect and exert TG-lowering effect by reducing VLDL secretion. (14)C-Pitavastatin is metabolized with CYP2C9 to 8-hydroxy derivative, but its Vmax /Km was about 2 micro l/min/mg, about 1/8 to 1/100 in comparison to the reported values of other statins, indicating that pitavastatin is hardly metabolized. Also, other human P450 species were not inhibited by pitavastatin. Therefore, pitavastatin is considered to have little interaction with drugs through P450. In the summarized clinical results with 862 patients, pitavastatin lowered TC and LDL-C by 28% and 40%, respectively. There was no difference in the frequency of side effects and no serious adverse effect was observed for pitavastatin. Pitavastatin possesses superior plasma lipid-improving effects, induces little drug interaction, and is expected to make a good contribution to the medication of hypercholesterolemia. PMID- 15118260 TI - [Effect of 4-ethylamino-2-butynyl(2-cyclohexyl-2-phenyl) glycolate, metabolite of oxybutynin, on intra-artery administered acetylcholine-induced urinary bladder contraction in anesthetized dogs]. AB - Oxybutynin has been used for neurogenic bladder disorders in clinic and known to have anti-cholinergic and spasmolytic properties. Metabolite of oxybutynin, 4 ethylamino-2-butynyl(2-cyclohexyl-2-phenyl) glycolate (N-desethyloxybutynin: DEOB) has been known to have similar anti-cholinergic and spasmolytic properties. However, the effect of DEOB on the urinary bladder has not been clarified in situ. Therefore, in the present study, we studied the effect of DEOB on acetylcholine-induced urinary bladder contraction in comparison with oxybutynin in anesthetized dogs. Intravenously administered DEOB dose-dependently inhibited acetylcholine-induced contractions. Oxybutynin also showed similar efficacy. From the Schild plot, it was found that the slope of DEOB and oxybutynin were 0.78 (95% confidence limit: 0.45-1.11) and 1.49 (95% confidence limit: 0.91-2.08), respectively. The dose of DEOB or oxybutynin needed to shift the concentration dependent curve of acetylcholine rightward to a two times higher dose was calculated. The doses of DEOB and oxybutynin were 6.4 micro g/kg (95% confidence limit: 1.7-12.8 micro g/kg) and 13.9 micro g/kg (95% confidence limit: 6.3-24.5 micro g/kg), respectively. From the above results, it was found that DEOB has the same anti-cholinergic property as oxybutynin and that its activity was almost equipotent to that of oxybutynin. Therefore, DEOB was suggested to play an important role during oxybutynin therapy for neurogenic bladder disorder. PMID- 15118261 TI - Onset age is associated with outcome of radioiodine therapy in Graves' disease. AB - Controversy remains regarding the optimal dose calculation with radioiodine therapy for patients with Graves' disease. Here, we focused our analysis on data concerning the patient's pretreatment background, an empirically set dose of radioiodine and the post-treatment thyroid function, and investigated those factors that affected the outcome. The subjects consisted of 38 patients diagnosed as having Graves' disease. All patients were hospitalized to undertake radioiodine therapy between 1989 and 1998 at our hospital. At the follow-up periods of 6-, 12-, and 36-months after therapy, we divided the patients into two groups: one group those who had hypothyroid function, and the other those who had normal or hyperthyroid function. At 6- and 12-months, 50% of the patients belonged to the hypothyroid function group, whereas at 36-months, 55% of them had hypothyroid function. Logistic regression analysis, with the objective variable being the post-treatment hypothyroidism after 12 months, revealed that the significant factor was the onset age. We suggested that the age at onset should be considered in patients with Graves' disease to determine the optional radioiodine dose for the therapy. PMID- 15118262 TI - K(ATP) channel knockout mice crossbred with transgenic mice expressing a dominant negative form of human insulin receptor have glucose intolerance but not diabetes. AB - Impaired insulin secretion and insulin resistance are thought to be two major causes of type 2 diabetes mellitus. There are two kinds of diabetic model mice: one is a K(ATP) channel knockout (Kir6.2KO) mouse which is defective in glucose induced insulin secretion, and the other is a transgenic mouse expressing the tyrosine kinase-deficient (dominant-negative form of) human insulin receptor (hIR(KM)TG), and which has insulin resistance in muscle and fat. However, all of these mice have no evidence of overt diabetes. To determine if the double mutant Kir6.2KO/hIR(KM)TG mice would have diabetes, we generated mutant mice by crossbreeding, which would show both impaired glucose-induced insulin secretion and insulin resistance in muscle and fat. We report here that: 1) blood glucose levels of randomly fed and 6 h fasted double mutant (Kir6.2KO/hIR(KM)TG) mice were comparable with those of wild type mice; 2) in intraperitoneal glucose tolerance test (ipGTT), Kir6.2KO/hIR(KM)TG mice had an impaired glucose tolerance; and 3) during ipGTT, insulin secretion was not induced in either Kir6.2KO/hIR(KM)TG or Kir6.2KO mice, while the hIR(KM)TG mice showed a more prolonged insulin secretion than did wild type mice; 4) hyperinsulinemic euglycemic clamp test revealed that Kir6.2KO, Kir6.2KO/hIR(KM)TG and hIR(KM)TG mice, showed decreased whole-body glucose disposal compared with wild type mice; 5) Kir6.2KO, but not Kir6.2KO/hIR(KM)TG mice had some obesity and hyperleptinemia compared with wild type mice. Thus, the defects in glucose-induced insulin secretion (Kir6.2KO) and an insulin resistance in muscle and fat (hIR(KM)TG) were not sufficient to lead to overt diabetes. PMID- 15118263 TI - Human growth hormone induces SOCS3 and CIS mRNA increase in the hypothalamic neurons of hypophysectomized rats. AB - The activation of the growth hormone (GH) receptor is followed by activation of the JAK2-STAT system in peripheral tissues, which in turn induces the expression of suppressors of cytokine signaling (SOCS) and/or cytokine-inducible SH2 protein (CIS) to achieve the attenuation of the signaling. To examine whether GH involves the SOCS/CIS system as intracellular negative regulators in the hypothalamus, we observed the effects of human GH on the gene expression of SOCS/CIS in the rat hypothalamus. The mRNAs of CIS, SOCS2, and SOCS3 in the hypothalamus of hypophysectomized male rats were examined by Northern analysis following the intravenous administration of recombinant human GH (hGH), 50 microg/100 g BW. The SOCS3 and CIS mRNAs were increased transiently with maximum expression at 1 h after hGH administration. The intravenous hGH did not induce SOCS2 mRNA expression in the hypothalamus. In situ hybridization demonstrated the increase of SOCS3 and CIS mRNAs in the arcuate nucleus after hGH administration, and the increase of SOCS3 mRNA in the periventricular nucleus. The hGH applied to primary cultured hypothalamic neurons at 500 ng/ml induced transient increase of SOCS3 and CIS mRNAs, but not SOCS2 mRNA. The results show that hGH acts directly on the neurons in the hypothalamus, and increases SOCS3 and CIS mRNAs, suggesting that these negative regulators may be involved in the mechanism that turns off the hGH action in the hypothalamic neurons. PMID- 15118264 TI - A rare case of Hurthle cell carcinoma with endobronchial metastasis. AB - Endobronchial metastases (EBM) are frequently seen in breast, renal and colon carcinomas. However, to our knowledge, only one case has ever been reported as EBM secondary to Hurthle cell carcinoma (HCC) in the literature. A 57-year-old woman had a bilateral total thyroidectomy for thyroid mass in 1990 that was diagnosed as HCC. She was admitted to our outpatient clinic in August 1999, with symptoms of cough, sputum, and right-sided pleuritic pain for the last seven months. In the bronchoscopic examination, two endobronchial lesions were seen. Pathological evaluation of the bronchoscopic samples was diagnosed as "Hurthle cell carcinoma" of thyroid. We suggest that, although rare, HCC should be considered in the differential diagnosis of the endobronchial metastasis. PMID- 15118265 TI - Effects of aging and postmenopausal hypoestrogenism on skin elasticity and bone mineral density in Japanese women. AB - Skin collagen content and bone mass decrease with aging. Loss of collagen from the skin might decrease its elasticity. We investigated associations between skin elasticity, bone mineral density (BMD), age, and menopausal hypoestrogenism. Thirty-eight healthy Japanese postmenopausal women were studied (mean age, 55.7 +/- 5.9 yr; range, 48 to 71). Skin elasticity was measured using a suction device applied to the dorsal right forearm. BMD values of L2 to 4 vertebral bodies were measured by dual-energy X-ray absorptiometry. Age showed significant negative correlations with both skin elasticity and BMD (r = -0.57, p<0.001 and r = -0.40, p<0.05, respectively). Years since menopause also showed significant negative correlations with both skin elasticity and BMD (r = -0.51, p<0.01 and r = -0.41, p<0.05, respectively). We also found a positive correlation between skin elasticity and BMD in these postmenopausal women (r = 0.44, p<0.01). In conclusion, we demonstrated declining skin elasticity and bone mass in postmenopausal women to possibly be age- and estrogen-related. Additionally, decreased skin elasticity might serve as a predictor of bone loss in postmenopausal women. PMID- 15118266 TI - Positive relationship between androgen and the endocrine disruptor, bisphenol A, in normal women and women with ovarian dysfunction. AB - This study was performed to investigate the serum levels of bisphenol A (BPA), an endocrine disruptor, in women with ovarian dysfunction and obesity. Fasting serum samples were obtained from 19 non-obese and 7 obese women with normal menstrual cycles: 7 patients with hyperprolactinemia, 21 patients with hypothalamic amenorrhea, and 13 non-obese and 6 obese patients with polycystic ovary syndrome (PCOS). BPA was measured by an enzyme-linked immunosorbent assay. BPA was detected in all human sera. Serum BPA concentrations were significantly higher in both non-obese and obese women with polycystic ovary syndrome (1.05 +/- 0.10 ng/ml, 1.17 +/- 0.16 ng/ml; p<0.05, respectively) and obese normal women (1.04 +/ 0.09 ng/ml, p<0.05) compared with those in non-obese normal women (0.71 +/- 0.09 ng/ml). There was no difference among women with hyperprolactinemia, women with hypothalamic amenorrhea, and non-obese normal women. There were significant positive correlations between serum BPA and total testosterone (r = 0.391, p<0.001), free testosterone (r = 0.504, p<0.001), androstenedione (r = 0.684, p<0.001), and DHEAS (r = 0.514, p<0.001) concentrations in all subjects. These findings show that there is a strong relationship between serum BPA and androgen concentrations, speculatively due to the effect of androgen on the metabolism of BPA. PMID- 15118267 TI - Effects of GABA and bicuculline on the electrical activity of rat olfactory placode neurons derived at E13.5 and cultured for 1 week on multi-electrode dishes. AB - The present study was performed to record the electrical activity of olfactory placode neurons and to check the effect of GABA and bicuculline on it. Olfactory placodes obtained at day 13.5 of gestation were cultured for 1 week on multi electrode dishes. Olfactory placode neurons showed spontaneous firing, with firing rates of 0.77 +/- 0.05 Hz (0.03-3.82 Hz, n = 12), but there was no bursting activity. Perfusion with 10 microM GABA almost immediately inhibited 8 of 11 firing activities (we could not test it in 1 activity). In contrast, perfusion with 10 microM bicuculline induced facilitation in 5 of 12 activities and did not induce any change in 7 other activities. Statistical analysis by chi(2)-test showed a significant difference in the response of neurons to the two drugs. Fisher's exact probability test showed that the inhibitory effect of GABA was significant (p<0.05) whereas neither the facilitatory effect nor the lack of effect of bicuculline was significant (p>0.1). These results suggest that cultured olfactory placode neurons, even in a probably immature stage, respond to GABA with inhibition, as generally observed at mature stages. PMID- 15118268 TI - Significant increase in maternal plasma leptin concentration in induced delivery: a possible contribution of pro-inflammatory cytokines to placental leptin secretion. AB - Maternal plasma leptin concentration is significantly increased during pregnancy. However, its roles in pregnancy, especially in labor, have not been fully clarified. We measured plasma leptin concentrations in pregnant women during the course of induced labor, just after spontaneous vaginal delivery and Cesarean section at term. We also studied the regulation of leptin secretion from term placental tissue and BeWo cells, a trophoblastic cell-line. Plasma leptin concentrations increased significantly during labor (58.9 +/- 9.2 ng/ml) compared to those before labor induction (37.5 +/- 5.8 ng/ml, P<0.05), then decreased 3-6 days postpartum (14 +/- 3 ng/ml, n = 6, P<0.0001) to the levels of normal nonpregnant women. Leptin concentrations within an hour and 24 hours after spontaneous vaginal delivery were significantly higher than those after Cesarean section (P<0.05 for both comparisons). Similarly, leptin mRNA expression in placental tissues obtained after spontaneous vaginal delivery was significantly greater than that in those obtained after Cesarean section without labor (P<0.05). IL-1alpha and TNF-alpha treatment significantly stimulated leptin secretion and leptin mRNA expression in explant culture of human term placental tissue and in BeWo cells as compared with those in vehicle controls (P<0.05, for all comparisons). By contrast, oxytocin and prostaglandin F(2alpha) treatment had no effects on leptin secretion from explant culture of human term placental tissue or from BeWo cells. These data indicate that pro-inflammatory cytokines might stimulate placental leptin secretion, thus finally contributing to the increase in plasma leptin concentration during labor. PMID- 15118269 TI - Constitutionally tall stature with morphological abnormality of the pituitary gland. AB - We describe two male pubertal cases of constitutionally tall stature (CTS) with an enlarged pituitary gland or pituitary microadenoma. Both patients' basal serum growth hormone (GH) levels were normal. Neither had oversecretion of any other pituitary hormones. However, their serum GH levels were paradoxically increased in response to an intravenous administration of thyrotropin-releasing hormone (TRH). A later GH response to an intravenous administration of luteinizing hormone-releasing hormone (LHRH) was also found in one patient. In addition, oral glucose loading (OGL) caused a late increase in serum GH at 180 min in both patients. Our present findings suggest that endocrinological and morphological abnormalities of the pituitary gland are found in some subjects with CTS during puberty. It is speculated that these paradoxical GH responses are associated with dysregulation of GH secretion. Finally, it is important to investigate whether tall children with morphological abnormalities of the pituitary gland such as our patients are preacromegalic, in which case, given that overproduction of GH secretion may occur in the future, further observation is necessary. PMID- 15118270 TI - Statural growth in 31 Japanese patients with SHOX haploinsufficiency: support for a disadvantageous effect of gonadal estrogens. AB - Although gonadal estrogens are known to facilitate the development of skeletal lesion in SHOX haploinsufficiency, controversy exists as to whether gonadal estrogens are disadvantageous to pubertal growth. To clarify this matter, we analyzed growth pattern in 31 Japanese patients with a normal karyotype and molecularly confirmed SHOX haploinsufficiency. The mean height SD score at the diagnosis of SHOX haploinsufficiency was similar between patients identified in childhood and those identified in adulthood (-2.7 +/- 0.8 [n = 15] vs. -2.4 +/- 0.7 [n = 16], P = 0.36), and was significantly lower in patients identified by the studies for short stature than in those ascertained by the familial studies of the probands both in childhood (-3.0 +/- 0.6 [n = 11] vs. -1.8 +/- 0.5 [n = 4], P = 0.0051) and in adulthood (-3.0 +/- 0.9 [n = 5] vs. -2.2 +/- 0.5 [n = 11], P = 0.040). Analysis of longitudinal paired growth data obtained in seven females showed a significantly different mean height SD score between childhood and adulthood (-2.3 +/- 0.5 vs. -2.9 +/- 0.8, P = 0.0060). The results imply that gonadal estrogens have a deleterious effect on pubertal growth in SHOX haploinsufficiency, and that the growth disadvantage is recognizable by longitudinal rather than cross-sectional growth studies. PMID- 15118271 TI - Reversible pituitary dysfunction in a patient with Cushing's syndrome discovered as adrenal incidentaloma. AB - We report a 45-year-old woman with Cushing's syndrome showing reversible pituitary dysfunction. Left adrenal tumor was incidentally discovered by a screening examination of abdominal computed tomography. Although this patient lacked typical Cushingoid features except hypertension and leg edema, endocrine examinations revealed moderate suppression of plasma ACTH (~6.3 pg/ml) with relatively high levels of serum cortisol (~22.9 microg/dl) without normal circadian rhythm. Plasma ACTH failed to respond to either CRH or metyrapone, and dexamethasone failed to suppress her daily steroid production. Surgical removal of left adrenocortical adenoma and 6-month replacement of hydrocortisone have ameliorated both ACTH and cortisol responses to CRH loading test. Postoperative responses of TSH and GH to TRH and GRH, respectively, were two fold higher than the preoperative levels. In contrast, basal and TRH-induced levels of serum PRL were decreased after surgery although both the basal and stimulated PRL levels were markedly high before surgery. In addition, gonadotropin response to GnRH examined in the same ovarian cycle was decreased in accordance with an increase in serum estradiol and progesterone levels after surgery. Improvement of hypercortisolemia even in a moderate case of Cushing's syndrome not only ameliorates hypertension, obesity and glucose intolerance, but also restores the accompanying dysfunctions of anterior pituitary, suggesting the clinical importance of early discovery and treatment of functioning adrenocortical incidentalomas. PMID- 15118272 TI - A rare case of primary hyperparathyroidism with clear cell adenoma. AB - We report a 52-year-old woman who was noted to have elevated alkaline phosphatase (ALP), hypercalcemia (Ca: 11.7 mg/dL), and intact parathyroid hormone (intact PTH: 643.1 pg/mL), and then referred to our hospital with suspected hyperparathyroidism. Ultrasound examination of the neck and magnetic resonance imaging showed a mass region in the posterior aspect of the left lobe of the thyroid, and Tl-Tc subtraction scintigraphy showed Tl uptake at the same location. Based on laboratory and imaging studies, she was diagnosed with primary hyperparathyroidism. The excised parathyroid was a large mass measuring 6.8 x 2.8 x 1.9 cm in diameter and weighing 15.4 g. It was soft, covered with a thin capsule, did not infiltrate the thyroid parenchyma, and showed no evidence of malignant process. Histopathological examination showed that it was clear cell adenoma. There was no evidence of metastasis from the parathyroid tumor in other organs. The post-operative course was excellent, and serum PTH, Ca, and ALP levels returned to normal. Among parathyroid tumors, large adenomas are commonly considered to be more likely malignant, but in this case it was benign despite measuring more than 6 cm in diameter. The histopathological type of the adenoma was clear cell adenoma, a very rare type. We report a clear cell adenoma of the parathyroid gland, which has not been described previously in Japan. PMID- 15118273 TI - Transient hyperthyroidism after withdrawal of antithyroid drugs in patients with Graves' disease. AB - The development of silent thyroiditis in patients with a history of Graves' disease is common, especially in the postpartum period. We describe herein patients with Graves' disease who developed transient hyperthyroidism but not silent thyroiditis after withdrawal of antithyroid drug (ATD). If such patients are diagnosed as recurrence of Graves' disease, they may receive ATD or radioiodine therapy unnecessarily. We investigated the characteristics of these patients to prevent unnecessary therapy. We retrospectively studied 22 patients with Graves' disease who showed transient thyrotoxicosis after withdrawal of ATD. Two of 22 patients were male and the mean ages (+/- SD) were 33.7 +/- 12.6 yr. We observed these patients for 28.5 +/- 12.8 (mean +/- SD; range 12-53) months after transient thyrotoxicosis, and measured TSH, FT4, and TSH binding inhibitor immunoglobulin in sera. Radioiodine uptake was measured in 6 of them. The radioiodine uptake in the 4 patients was not suppressed (27.5%, 28.0%, 32.7%, 38.1%). These uptake levels indicate that their thyrotoxicosis was not caused by silent thyroiditis. Most of the 22 patients became euthyroid within 6 months. This study suggests a new therapeutic option as follows: in the case of young patients with mild thyrotoxicosis after withdrawal of ATD, physicians should follow them up for one month without medication unless they have unbearable symptoms or complications. PMID- 15118274 TI - Follicular thyroid carcinomas with lung metastases: a 23-year retrospective study. AB - Limited clinical information is specified in the presentations, results of treatment and prognostic factors of follicular thyroid carcinoma with lung metastases. In order to better characterize the information, we retrospectively analyzed the data of 2,003 thyroid cancer patients who received treatment and follow-up at the Chang Gung Memorial Hospital during the period from January 1979 to December 2002. There were 1,516 cases of papillary and 272 cases of follicular thyroid carcinomas. In the study, lung metastases of the follicular thyroid carcinomas were defined as post-operative or follow-up chest X-ray, diagnostic or therapeutic (131)I scan with positive finding of lung metastases. Serum thyroglobulin (Tg) levels under thyroxine treatment of patients with lung metastases had to be over 1.5 ng/mL. Of the follicular thyroid carcinomas, there were 70 (25.7%) with lung metastases including 50 females (mean age 54.1 +/- 12.6 years old) and 20 males (mean age 59.4 +/- 12.0 years old). Of the 70 patients, there were 53 patients (75.7%) who presented with lung metastases at the time of diagnosis. Of the 70 patients of follicular thyroid carcinoma with lung metastases, 30 patients (42.9%) died at the end of the follow-up, and only 4 patients improved to disease free status. The 5, 10, 15, and 20 year survival rates in these patients were 68.5%, 54.0%, 41.6%, 27.7%, respectively. Age, post operative Tg level and tumor size are important prognostic factors which are demonstrated to be significantly different statistically between lung metastases group and the group of the patients without distant metastasis. Otherwise, only the tumor size and accumulative dose of (131)I therapy demonstrate a significant difference between survival and mortality groups. Seventeen of the 70 patients developed lung metastases during the follow-up period. Mean period between diagnosis and recurrence of these patients was 3.6 +/- 0.9 years. Over 75% of follicular thyroid carcinoma with lung metastases was diagnosed at the time of presentation. Forty percent (28/70 cases) of the follicular thyroid carcinoma with lung metastases had history of thyroid surgery in this study. If lung metastases are diagnosed in follicular thyroid carcinoma, it will be followed by a poor prognosis. Older patients, higher postoperative Tg, and larger tumor size in follicular thyroid carcinoma need aggressive postoperative treatment. PMID- 15118275 TI - Somatostatin (SRIF) receptor subtype 2 and 5 gene expression in growth hormone secreting pituitary adenomas: the relationship with endogenous srif activity and response to octreotide. AB - To investigate the potential pathophysiologic role of human SRIF receptor gene expression in GH-secreting adenomas in acromegalic patients, we studied the relationship between the SRIF receptor gene expression, endogenous SRIF activity and exogenous response to octreotide in 16 acromagalic patients. Hypothalamic somatostatinergic activity (HSA) was assessed by glucose-induced suppression of TRH-stimulated TSH secretion. As an indicator of somatotrope sensitivity to HSA, glucose-induced suppression of TRH-stimulated GH secretion was determined. For the acute octreotide response, a 100 microg bolus of octreotide was injected intravenously and GH was measured hourly for 6 hr. Pituitary tumor SRIF receptor subtype 2 and 5 (sst2 and sst5) mRNA levels were measured by real-time RT-PCR. Gsp oncogene was also detected by direct PCR sequencing. Sst2 and sst5 mRNA levels were detected in all tumors. Sst2 mRNA levels positively correlated with that of sst5. Sst2 and sst5 mRNA levels did not show any correlation with basal GH values (nadir or peak). Expression of sst2, but not sst5, showed a positive correlation with the GH response to HSA, while the octreotide response positively correlated with the sum of sst2 and sst5 mRNA levels. Individuals with gsp positive tumors were more responsive to octreotide than those with gsp-negative tumors but sst2 and sst5 mRNA levels did not differ between these two groups. These results suggest common transcriptional and/or post-transcriptonal regulatory mechanisms for these SRIF receptor subtypes within GH-secreting pituitary adenomas. The functional observations suggest that the degree (or level) of sst2 and sst5 expression is critical for the ultimate GH response of somatotropinomas to endogenous SRIF tone and exogenous SRIF analogue therapy. However, sst2 and sst5 mRNA levels are not the only factors mediating the response to SRIF. PMID- 15118276 TI - A case of malignant thymoma mimicking thyroid carcinoma: a pitfall in fine-needle aspiration. AB - A case of malignant thymoma presenting as an anterior neck mass is reported. The tumor extended from the thyroid gland to the superior mediastinum. It did not accumulate Tc-99m pertechnetate, but continued to accumulate Tl-201 at the late phase. A fine-needle aspiration cytology from the tumor showed tight clusters of epithelial cells with crowded ovoid nuclei. The tumor was initially diagnosed as thyroid carcinoma, clinically and cytologically. A thymoma with a dominant epithelial component has to be considered in the differential diagnosis of a suspected papillary carcinoma of the thyroid. PMID- 15118277 TI - Aldosterone stimulates gene expression of hepatic gluconeogenic enzymes through the glucocorticoid receptor in a manner independent of the protein kinase B cascade. AB - Primary aldosteronism is associated with glucose intolerance and diabetes, which is due in part to impaired insulin release caused by reduction of potassium, although other possibilities remain to be elucidated. To evaluate the in vivo effects of aldosterone on glucose metabolism, a single dose of aldosterone was administered to mice, which resulted in elevation of the blood glucose level. In primary cultured mouse hepatocytes, the gene expression of gluconeogenic enzymes such as glucose-6-phosphatase (G6Pase), fructose-1,6-bisphosphatase and phosphoenolpyruvate carboxykinase increased in response to aldosterone in a dose dependent manner even at a concentration similar to a physiological condition (10(-9) M). The inhibitory effect of insulin on G6Pase gene expression was partially suppressed by aldosterone. Furthermore, aldosterone enhanced G6Pase promoter activity in human hepatoma cell line HepG2, which was prevented by co treatment with a glucocorticoid antagonist RU-486, but not a mineralocorticoid antagonist spironolactone. In contrast, aldosterone had no effects on major insulin signaling pathways including insulin receptor substrate-1, protein kinase B, and forkhead transcription factor. These results suggest that aldosterone may affect the inhibitory effect of insulin on hepatic gluconeogenesis through the glucocorticoid receptor, which may be one of the causes of impaired glucose metabolism in primary aldosteronism. PMID- 15118278 TI - The importance of communication between multiple physicians treating the same patient. PMID- 15118279 TI - Relation of serum total cholesterol and other risk factors to risk of coronary events in middle-aged and elderly Japanese men with hypercholesterolemia: the Kyushu Lipid Intervention Study. AB - BACKGROUND: The role of serum total cholesterol (TC) in the development of coronary heart disease (CHD) may differ in different age groups. METHODS AND RESULTS: The relation of serum TC and other risk factors to CHD events was examined in middle-aged (<65 years) and elderly (> or =65 years) men separately in the Kyushu Lipid Intervention Study (KLIS). Subjects were 4,349 men aged 45-74 years with serum TC of 220 mg/dl or greater who had no history of myocardial infarction, coronary angioplasty, or stroke. There were 123 CHD events (ie, myocardial infarction, coronary bypass surgery, coronary angioplasty, cardiac death, and sudden death) in a 5-year follow-up period. The Cox proportional hazards model was used with baseline and follow-up serum TC, baseline high density lipoprotein (HDL) cholesterol, hypertension, diabetes mellitus, and other factors as covariates. Serum TC concentration during the follow-up, not at baseline, was associated with an increased risk of CHD events, especially in elderly men. High concentrations of serum HDL cholesterol were associated with a modest, statistically nonsignificant decrease in the risk among middle-aged men. An increased risk of CHD events associated with diabetes mellitus was greater in middle-aged men. Hypertension and smoking were not measurably related to the risk in either middle-aged or elderly men. CONCLUSIONS: Both the serum TC concentration during follow-up and diabetes mellitus are important predictors of CHD events in Japanese men with moderately elevated serum TC. PMID- 15118280 TI - Aspirin attenuates the incidence of silent brain lesions in patients with nonvalvular atrial fibrillation. AB - BACKGROUND: Abnormal findings, including silent cerebral infarction, are frequently observed by magnetic resonance imaging (MRI) in patients with nonvalvular atrial fibrillation (NVAF); however, the prevalence and prevention strategy for these lesions have not been extensively studied. In the present study the preventive effects of aspirin on silent ischemic lesions was investigated. METHODS AND RESULTS: Silent lesions were counted using cranial MRI performed in 78 neurologically normal adults with sinus rhythm and in 212 patients with NVAF without a history of stroke. MRIs were repeated twice in the NVAF patients at 12-month intervals. During the first year, patients received neither antiplatelet agent nor anticoagulant; in the second year, aspirin (330 mg daily) was administered. The prevalence of lesions in the initial MRI was higher in NVAF patients (86.4%) than in sinus rhythm subjects (53.8%; p<0.001). After 12 months without aspirin, new lesions were seen in 20.6% of NVAF patients. The yearly occurrence of new lesions was decreased to 9.6% during the year of treatment with aspirin (p=0.014). CONCLUSIONS: In patients with NVAF, abnormal lesions are frequently observed by MRI and aspirin treatment may be effective in preventing further small silent lesions. PMID- 15118281 TI - Attitudes of Japanese cardiologists toward anticoagulation for nonvalvular atrial fibrillation and reasons for its underuse. AB - BACKGROUND: Although warfarin reduces embolic events in patients with nonvalvular atrial fibrillation (NVAF), it is used less frequently in Japan and so the aim of the present study was to determine the attitudes of Japanese cardiologists toward antithrombotic therapy for NVAF patients. METHODS AND RESULTS: Subjects were NVAF patients enrolled in a prospective study in 1999. Clinical characteristics, type of NVAF and antithrombotic therapy, risk factors for embolism, and contraindications to warfarin were analyzed. Risk factors included advanced age (>75 years), hypertension, diabetes mellitus, congestive heart failure, and prior embolic events. Contraindications to warfarin included bleeding tendency, malignant tumors and others. Among 509 patients (66.6+/-10.3 years old), 359 had at least one risk factor for embolism and of these 359 patients, 200 (55.7%) received warfarin (ie, modest adherence to the guideline for antithrombotic therapy). There were 159 patients who had at least one risk factor but did not receive warfarin; 70.4% of these received antiplatelet drugs. Contraindications were found in only 22.6% and paroxysmal nature of NVAF seemed a possible reason for non-use of warfarin in 47.2% of 159 patients. CONCLUSIONS: In Japan warfarin is not used extensively for treatment of NVAF patients having risk factors and the reasons for not using antithrombotic therapy seemed inappropriate in most of patients. PMID- 15118282 TI - Effect of reperfusion therapy on cardiac rupture after myocardial infarction in Japanese. AB - BACKGROUND: Cardiac rupture after acute myocardial infarction (AMI) is unpredictable and almost always fatal, so the present study reviewed all the clinical characteristics of patients with cardiac rupture to determine if its occurrence can be predicted. METHODS AND RESULTS: The clinical characteristics of 1,296 consecutive AMI patients from January 1992 to February 2002 were retrospectively evaluated using multi-logistic analysis. Cardiac rupture occurred in 45 patients (3.5%), comprising left ventricular free wall rupture (n=23), ventricular septal perforation (n=20) and papillary muscle rupture (n=2). Early phase rupture (within 72 h after AMI onset) was associated with anterior infarction. Of the 45 patients who experienced ruptures, 24 (53.3%) survived to discharge; 21 (46.6%) of the 45 ruptures occurred after admission. Successful reperfusion therapy was performed within 24 h for 840 patients. The incidence of rupture was significantly lower for reperfused patients than for non-reperfused patients (0.6% vs 3.5%, respectively; p<0.01). Peak C-reactive protein concentration was a reliable predictor of late-phase rupture (p=0.01), but not of early-phase rupture. CONCLUSIONS: Reperfusion therapy appears to aid in the prevention of cardiac rupture, especially late-phase rupture. PMID- 15118283 TI - Analysis of chronic heart failure registry in the Tohoku district: third year follow-up. AB - BACKGROUND: Because the real prognosis of Japanese patients with chronic heart failure (CHF) is still unknown, the aim of the present study was to clarify the prognosis and predictors for mortality of CHF patients using the Chronic Heart failure Analysis and Registry in Tohoku district (CHART). METHODS AND RESULTS: As of February 2003, 1,154 stable CHF patients with optimum standard therapy have been enrolled in the registry since February 2000 and of these, 175 died of some cause during the follow-up period (mean follow-up period, 1.9+/-0.9 years), giving 1- and 3-year all-cause mortality rates of 7.3% and 20.9%, respectively. Multivariate analysis showed that age at entry, diabetes, ventricular tachycardia, plasma concentration of brain natriuretic peptide (BNP), New York Heart Association (NYHA) functional class and rural residence were significantly associated with all-cause mortality (hazard ratio: 1.028, 1.940, 1.650, 1.001, 1.713, and 2.226). The 1-year all-cause mortality rates of CHF patients with a left ventricular ejection fraction <25%, left ventricular end-diastolic diameter > or =60 mm, BNP > or =500 pg/ml, NYHA > or = III, or with underlying coronary artery disease were 15.0%, 11.4%, 16.8%, 16.3%, and 10.8%, respectively. CONCLUSIONS: The prognosis of Japanese CHF patients with these predictors remains poor. A stratified approach is necessary to improve their survival and quality of life. PMID- 15118284 TI - Influence of CD36 deficiency on heart disease in children. AB - BACKGROUND: The physiological role of the CD36 molecule in pediatric heart disease has not been fully investigated. METHODS AND RESULTS: The CD36 antigen in platelets and monocytes was measured by flow cytometry in 189 patients with various heart diseases; 15 (7.9%) had a diagnosis of CD36 deficiency (type I: 2[1 boy, 1 girl], type II: 13 [6 boys, 7 girls]). The prevalence in each heart disease was as follows: group A (congenital heart disease) 7.6% (9/118, type II: 9 [6 boys, 3 girls]); group B (myocardial disease) 20.0% (3/15, I: 1 girl, II: 2[1 boy, 1 girl]), group C (Kawasaki disease) 4.9% (2/41, II: 2 [1 boy, 1 girl]), group D (arrhythmia): 6.7% (1/15, I: 1 boy). Three patients in group B had transient myocardial damage, which was thought to be related to abnormal myocardial long-chain fatty acid metabolism. CONCLUSION: The frequency of CD36 deficiency in childhood heart disease was almost identical to that of healthy individuals. Some patients with CD36 deficiency may be susceptible to myocardial damage in the presence of disadvantageous conditions, such as serious infections or massive steroid therapy. PMID- 15118285 TI - Pathogenetic significance of myxomatous degeneration in fenestration-related massive aortic regurgitation. AB - BACKGROUND: Aortic valvular regurgitation has several mechanisms and the present study investigated its clinicopathological correlations with aortic valve fenestration. METHODS AND RESULTS: Six male patients with massive regurgitation and enlarged fenestrations or ruptured fenestrated fibrous cords underwent aortic valve replacement. The clinicopathological features showed many similarities. Four cases had family histories of aortic regurgitation. All six patients showed moderate to severe myxomatous degeneration of the aortic valve and enlargement of aortic annulus. Four patients had 1-2 ruptured fibrous cords, located at the right coronary cusp. Echocardiographic examinations showed an abnormal fibrous cord attached to the prolapsing cusp in 3 cases with ruptured fenestrated valve and detailed examination by transesophageal echocardiography showed an intact fenestrated fibrous cord at the commissure in 1 case. CONCLUSION: Extensively fenestrated cusps with an enlarged aortic annulus because of myxomatous degeneration can cause chronic regurgitation or sudden deterioration after rupture of the fibrous cord. There is an increasing incidence of fenestration related aortic regurgitation in the Japanese population. An important pathogenetic factor in male patients is myxomatous degeneration of the aortic cusp and annulus. PMID- 15118286 TI - Neurohumoral profiles in patients with hypertrophic cardiomyopathy: differences to hypertensive left ventricular hypertrophy. AB - BACKGROUND: Patients with hypertrophic cardiomyopathy (HCM) or hypertensive heart disease (HHD) have increased concentrations of various neurohumoral factors. Thus, the aim of the present study was to evaluate the differences in the neurohumoral profiles of HCM and HHD. METHODS AND RESULTS: Plasma concentrations of epinephrine, norepinephrine, atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), angiotensin II and endothelin-1 were measured in 40 patients with HCM, 35 with HHD, and 15 controls. Additionally, the concentrations of these neurohumoral factors in the coronary sinus and aortic root were measured in 12 HCM patients and 10 controls. Plasma concentrations of norepinephrine, ANP and BNP were significantly higher in HCM than HHD and controls. In HCM, there was no significant correlation between the left ventricular mass index and any neurohumoral factor. The plasma BNP concentration significantly correlated with left intraventricular pressure gradient in HCM. There were significant differences in the plasma concentrations of ANP and BNP between HCM with and without left ventricular diastolic dysfunction. Transcardiac production of BNP was significantly higher in patients with obstructive HCM than in those with non obstructive HCM. CONCLUSIONS: The significant neurohumoral differences between HCM and HHD were the plasma concentrations of norepinephrine, ANP and BNP. In HCM patients, the plasma BNP concentration may reflect the intraventricular pressure gradient and left ventricular diastolic dysfunction whereas the plasma ANP concentration reflects only the left ventricular diastolic dysfunction. PMID- 15118287 TI - Increases in interleukin-6 and matrix metalloproteinase-9 in the infarct-related coronary artery of acute myocardial infarction. AB - BACKGROUND: To elucidate the involvement of inflammation in coronary artery occlusion, the regional changes in cytokines and matrix metalloproteinases (MMPs) in the infarct-related coronary artery were determined in patients with acute myocardial infarction. METHODS AND RESULTS: Cardiac catheterization was carried out within 24 h of the onset of infarction in 36 patients. Blood samples were collected from the infarct-related coronary artery, the ascending aorta and the peripheral vein. Plasma interleukin (IL)-6 concentrations were elevated in all 3 samples. Particularly, the plasma IL-6 concentrations were 14.4 pg/ml in the infarct-related coronary artery, a value significantly greater than the respective 8.0 pg/ml and 6.5 pg/ml in the ascending aorta and peripheral vein. Plasma IL-6 concentrations in the infarct-related coronary artery had positive correlation with all of the following references measured by intravascular ultrasound: external elastic membrane cross-sectional area (EEM-CSA) (r=0.47, p<0.01), lesion EEM-CSA (r=0.51, p<0.01) and plaque area (r=0.48, p<0.01). MMP-9 was increased regionally in the infarct-related coronary artery, at 11.8 ng/ml vs 8.2 ng/ml in the ascending aorta (p<0.001). CONCLUSIONS: The present findings suggest that IL-6 at least is released from the ruptured vulnerable atherosclerotic plaque and that regional activation of macrophages is involved in the occlusive process of coronary artery in acute myocardial infarction. PMID- 15118288 TI - Stenting after directional coronary atherectomy compared with directional coronary atherectomy alone and stenting alone: a serial intravascular ultrasound study. AB - BACKGROUND: Directional coronary atherectomy prior to stent implantation (DCA stent) is expected to be an effective approach to reduce restenosis. The purpose of this study was to determine whether DCA-stent has advantages over DCA alone or stenting alone using serial intravascular ultrasound (IVUS). METHODS AND RESULTS: Serial (pre-, post- and follow-up) IVUS was performed in 187 native coronary lesions treated with each of the 3 strategies. External elastic membrane cross sectional area (CSA), lumen CSA and plaque CSA were measured. Baseline characteristics were similar. Postprocedural lumen CSA was largest after DCA stent (11.2+/-2.7 mm2) and DCA (10.8+/-2.5 mm2) than stenting alone (9.0+/-2.9 mm2) (p<0.0005). Follow-up lumen loss was similar. As a result, follow-up lumen CSA was largest after DCA-stent (DCA-stent: 9.1+/-3.4 mm2, DCA: 7.8+/-4.2 mm2, stent: 6.3+/-2.6 mm2, p<0.0005). There was a trend toward a lower rate of restenosis with DCA-stent (DCA-stent, 12.5%; DCA, 18.3%; stent, 18.8%; p=0.57). CONCLUSIONS: DCA-stent is superior to both DCA alone and stent alone in terms of the ability to gain a larger lumen as assessed by IVUS. PMID- 15118289 TI - Usefulness of stress myocardial perfusion imaging for evaluating asymptomatic patients after coronary stent implantation. AB - BACKGROUND: Stent implantation in coronary angioplasty has reduced the rate of restenosis, but many patients still undergo follow-up coronary angiography (CAG). The present study was a multi-center retrospective analysis of the usefulness of stress single photon emission computed tomography (SPECT) compared with follow-up CAG in stent-implanted patients who remained asymptomatic during the follow-up period. METHODS AND RESULTS: The study group of 103 patients underwent both SPECT and CAG at 4-9 months after stent implantation. Restenosis occurred in 20 (19%) of 106 vessel territories, and a reversible perfusion defect was found in 32 (30%) territories. Sensitivity, specificity, positive and negative predictive values, and accuracy of SPECT were 65%, 78%, 41%, 91%, and 76%, respectively. The accuracy was lower in territories with a prior myocardial infarction (71%), in the left circumflex artery (58%), and in cases with three-vessel disease (63%). The negative predictive value was high, but 7 false negative cases included 4 cases with prior myocardial infarction, and 2 cases with reversible defects in other vessel territories. CONCLUSIONS: Stress SPECT imaging is a useful tool for following up patients with coronary stent implantation, and follow-up CAG could be omitted in patients with negative SPECT imaging, no prior myocardial infarction, one- or two-vessel disease, and sufficient stress loading. PMID- 15118290 TI - Minimally-diluted blood cardioplegia supplemented with potassium and magnesium for combination of 'initial, continuous and intermittent bolus' administration. AB - BACKGROUND: The present study was designed to examine the hypothesis that minimally-diluted blood cardioplegia (BCP) supplemented with potassium and magnesium provides superior myocardial protection in comparison with the standard diluted BCP for a combination of 'initial, continuous, and intermittent bolus' BCP administration. METHODS AND RESULTS: Seventy patients undergoing elective coronary revascularization between 1997 and 2001 (M : F =55:15, mean age 67.6+/ 7.5 years) were randomly divided into 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 min of initial antegrade BCP infusion, followed by continuous retrograde BCP infusion. Intermittent antegrade BCP was infused every 30 min for 2 min. 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). The number of patients showing spontaneous heart beat recovery after reperfusion was significantly larger in Group M (28 vs 15, p=0.0029), and the number of patients suffering from atrial fibrillation during the postoperative period was significantly smaller in Group M (n=3 vs 11, p=0.034). Both the postoperative maximum dopamine dose (3.57+/-2.46 vs 5.44+/-2.23 microg/kg per 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 less in Group M. The number of patients showing paradoxical movement of the ventricular septum in the early postoperative echocardiography was significantly smaller in Group M (9 vs 24, p=0.0007). CONCLUSIONS: These results suggest that 'initial, continuous and intermittent bolus' administration of minimally-diluted BCP supplemented with potassium and magnesium is a reliable and effective technique for intraoperative myocardial protection. PMID- 15118291 TI - Sinus node function in patients with Brugada-type ECG. AB - BACKGROUND: Some studies have shown that patients with Brugada syndrome (BS) have atrioventricular conduction disturbance, but their sinus node function has not been evaluated. METHODS AND RESULTS: The patients group consisted of 59 male patients and 1 female patient with BS. Supraventricular and ventricular programmed electrical stimulation (PES) was performed. Ventricular fibrillation (VF) or sustained polymorphic ventricular tachycardia was induced by ventricular PES in 26 patients with BS (VF group), but was not induced in the other 34 patients (non-VF group). Sinus node function and conduction of the atrioventricular (AV) node in the control group, non-VF group and VF group were evaluated. Sinus node function was attenuated and the His - ventricle interval was prolonged in the VF group (corrected sinus node recovery time: 452+/-126 ms (VF group), 324+/-146 ms (non-VF group), Sino-atrial conduction time: 179+/-60 ms (VF group), 127+/-60 ms (non-VF group), His-ventricle interval: 41+/-9 ms (VF group), 35+/-8 ms (non-VF group)). CONCLUSION: The function of both the sinus node and AV node are attenuated in patients with PES-induced VF. PMID- 15118292 TI - B-type natriuretic peptide as a marker of resuscitation in patients with cardiac arrest outside the hospital. AB - BACKGROUND: Although the circulating concentration of B-type natriuretic peptide (BNP) has both a prognostic and diagnostic value in heart disease, no data are available regarding its resuscitative value for out-of-hospital cardiac arrest. METHODS AND RESULTS: The present study was a prospective study of 401 patients whose BNP was measured on arrival at the emergency room after an out-of-hospital cardiac arrest with a cardiac cause. The primary endpoint was survival to hospital discharge. The unadjusted rate of survival to hospital discharge decreased in a stepwise fashion among patients in increasing quartiles of BNP concentration (p<0.001). After adjusting for independent predictors of resuscitation, the odds ratios for survival to hospital discharge in the second, third and fourth quartiles of BNP were 0.13 (95% confidence interval (CI), 0.04 0.46), 0.10 (95% CI, 0.03-0.41), and 0.004 (95% CI, 0.00-0.16), respectively. The BNP cutoff value of 100 pg/ml for survival had a sensitivity of 83% and a negative predictive value of 96%. CONCLUSIONS: The measurement of BNP was found to provide valuable predictive information for survival to hospital discharge in patients with out-of-hospital cardiac arrest of cardiac etiology. PMID- 15118293 TI - Diagnosis of multivessel coronary vasospasm by detecting postischemic regional left ventricular delayed relaxation on echocardiography using color kinesis. AB - BACKGROUND: It is not known whether multivessel coronary spasm occurs spontaneously in patients who have variant angina (VA) with demonstrated multivessel spasm induced by intracoronary injection of acetylcholine (ACh). Regional left ventricular (LV) diastolic dysfunction or wall motion abnormality may persist after an episode of coronary vasospasm. Color kinesis (CK) is a recent development that facilitates the echocardiographic evaluation of regional diastolic wall motion. METHODS AND RESULTS: Regional diastolic wall motion was evaluated using CK in 26 patients with VA within 1 week of the last episode of angina. The LV segmental filling fraction in the short-axis view during the first 30% of the diastolic filling time, expressed as a percentage, was used to objectively identify postischemic diastolic endocardial motion asynchrony. Diastolic asynchrony or regional LV delayed relaxation was noted in all 26 (100%) patients and in 14 (54%) it was detected in multiple vascular territories, suggesting multivessel spasm. Multivessel spasm was induced by ACh in 11 (79%) of the patients with suspected multivessel spasm by CK. In 11 (92%) of the 12 patients with multivessel spasm induced by ACh multiple regions of delayed relaxation had been noted by CK. The regions of delayed relaxation were largely consistent with the territories perfused by the arteries reacting to ACh (sensitivity: 96%, specificity: 91%). CONCLUSION: ACh induced spasm in the same coronary arteries as those perfusing the regions with delayed diastolic wall motion detected by CK in most of the patients with VA, suggesting that multivessel spasm does occur spontaneously in patients with susceptible arteries. PMID- 15118294 TI - Vascular endothelial growth factor is an autocrine growth factor for cardiac myxoma cells. AB - BACKGROUND: Cardiac myxomas are generally considered benign, but malignant tumors have been reported. Vascular endothelial growth factor (VEGF), an angiogenic factor, plays a role in the growth, progression, and metastasis of solid tumors and it has been reported that VEGF expression is upregulated in cardiac myxomas that have a high microvessel density. The purpose of this study was to determine whether cardiac myxoma cells possess a VEGF-autocrine system that regulates tumor growth. METHODS AND RESULTS: Immunohistochemical analyses revealed the presence of VEGF and its receptors, VEGFR-1 (flt-1) and VEGFR-2 (KDR/flk-1), in the cytoplasm of tumor cells from 18 of 18 myxoma tissue specimens examined. Two different myxoma cell lines were established and constitutively secreted large amounts of VEGF as determined by enzyme-linked immunosorbent assay. The expression of VEGF, VEGFR-1, and VEGFR-2 mRNA was detected in both cell lines by reverse-transcriptase polymerase chain reaction. Myxoma cell proliferation, as determined by thymidine incorporation, was enhanced by the addition of VEGF in a dose-dependent manner, and cell proliferation was inhibited in a dose-dependent manner by the addition of a neutralizing VEGF antibody. CONCLUSIONS: These results indicate that cardiac myxoma cells possess a VEGF-autocrine system, which could contribute to the malignant potential of histologically benign myxomas through direct stimulation of tumor cell growth as well as through induction of angiogenesis. PMID- 15118295 TI - Oral verapamil attenuates the progression of pacing-induced electrical and mechanical remodeling of the atrium. AB - BACKGROUND: Calcium overload plays a major role in the development of electrical and mechanical remodeling during atrial fibrillation, but the potential of verapamil, a Ca blocker, for preventing atrial electrical remodeling remains controversial. METHODS AND RESULTS: Pacing and recording electrodes were sutured to the right atrium in 16 dogs. After a 5-day recovery period, rapid atrial pacing at 400 ppm was initiated in 8 dogs (control group). In the remaining 8 dogs, oral administration of verapamil (8 mg/kg per day) was started 1 week before the initiation of rapid pacing (verapamil group). On the day before and at 2, 7, 14 days after rapid pacing, electrophysiological (EP) and transesophageal echocardiographic (TEE) studies were performed under autonomic blockade. In response to rapid pacing, EP and TEE parameters changed progressively in the control group (p<0.05 vs day 0), whereas in the verapamil group, no significant changes in the various parameters were observed for the first 7 days. However, verapamil failed to prevent progression of both types of remodeling after 14 days of pacing. CONCLUSION: Verapamil can attenuate the progression of electrical and mechanical remodeling of the atrium for at least 7 days. PMID- 15118296 TI - Successful pregnancy in a patient with double outlet left ventricle after a Rastelli operation using a prosthetic valve. AB - A woman with double outlet left ventricle (DOLV) had undergone a Rastelli operation using a prosthetic Bjork Shiley valve and who was receiving anticoagulant drug delivered a healthy male infant. Oral warfarin was replaced by heparin from the 5th to the 13th week of gestation and for the last 5 weeks of gestation. Successful pregnancy in patients with DOLV after a Rastelli operation using a prosthetic valve is possible with careful maintenance. PMID- 15118297 TI - Double primary left ventricular and aortic valve papillary fibroelastoma. AB - Papillary fibroelastomas of the heart are relatively rare benign tumors and although they can be symptom free, symptoms such as cerebral ischemia and cardiac infarction can be lethal. It is important to diagnose this tumor using echocardiography, because lethal embolisms can be prevented by surgical resection. Recurrence of this tumor has not been reported and multiple tumors are rare. The present report is a case of double primary aortic valve fibroelastoma with aortic regurgitation diagnosed 4 years after surgery for a left ventricular fibroelastoma with mitral valvular disease. PMID- 15118298 TI - Replacement of an aortic Starr-Edwards ball valve prosthesis 28 years after implantation. AB - A 53-year-old woman who had undergone aortic valve replacement with a Starr Edwards (S-E) valve (Model 1260) and open mitral commissurotomy 28 years previously was hospitalized with cardiac failure. Echocardiography showed mitral stenosis, mitral regurgitation, and a normally functioning S-E prosthesis. At reoperation, the mitral and aortic valves were replaced with St Jude bileaflet mechanical prostheses. Examination of the explanted S-E prosthesis revealed no structural abnormality other than lipid infiltration of the silastic ball. PMID- 15118299 TI - Gitelman's syndrome with exercise-induced ventricular tachycardia. AB - A 62-year-old female with palpitations was admitted to hospital where she recorded 12,299 monofocal ventricular premature contractions (VPCs) in 24 h and nonsustained ventricular tachycardia (VT) on exertion. She had hypokalemia with renal potassium wasting, a chloride-resistant metabolic alkalosis, elevated plasma renin, elevated plasma aldosterone (relative to the serum K concentration), hypomagnesemia with renal magnesium wasting, decreased urine calcium excretion, and normal blood pressure. The hypokalemia and hypomagnesemia were thought to have precipitated the VT. The coronary angiogram showed normal coronary arteries; however, the left ventriculogram revealed akinesis of the posterolateral wall. Because the VT could not be induced by programmed electrical stimulation either before or during intravenous administration of isoproterenol, the VPC with the same QRS morphology as the VT became the target of radiofrequency catheter ablation (RF-CA). Intracardiac mapping showed that the earliest activation site was situated in the asynergic area of the left ventricle (LV) and radiofrequency catheter ablation directed at the LV asynergy area completely eliminated the VPCs without any complications. During the follow-up period (6 months), she was free from palpitation and VT was not clinically documented. PMID- 15118300 TI - Multi-detector computed tomography for imaging of subendocardial infarction: prediction of wall motion recovery after reperfused anterior myocardial infarction. AB - BACKGROUND: After reperfusion therapy for acute myocardial infarction (AMI), evaluation of transmural myocardial microcirculation can indicate prognosis. The aim of the present study was to determine whether the evaluation of transmural myocardial microcirculation by newly developed 4-slice computed tomography (CT) can estimate the recovery of left ventricular function. METHODS AND RESULTS: Seventeen consecutive patients who had anterior AMI with a total occlusion in the proximal left anterior descending artery (LAD) and who had undergone successful balloon reperfusion therapy within 24 h of the onset of AMI were examined. Four slice CT was performed 10-14 days after AMI onset. The median of the epicardial perfusion ratio (infarcted anterior epicardial CT number/intact lateral epicardial CT number ratio = 92%) was used to categorize the cases into 2 groups: the transmural infarction group (n=8) and the subendocardial infarction group (n=9). Although no significant difference was observed between myocardial enhancement by CT in the acute phase and anterior wall motion or ejection fraction in the acute phase, the transmural infarction group showed poor recovery of anterior wall motion at 6 months after AMI onset, whereas the subendocardial infarction group exhibited good recovery of regional and global left ventricular function. CONCLUSIONS: Transmural myocardial microcirculation imaged by 4-slice CT can predict wall motion recovery after AMI. PMID- 15118301 TI - Identification of a collagen production-promoting factor from an extract of royal jelly and its possible mechanism. AB - We have previously shown that royal jelly (RJ) promoted collagen production by skin fibroblasts in the presence of ascorbic acid-2-O-alpha-glucoside (AA-2G). In this study, we purified the honeybee RJ-derived collagen production-promoting factor (HBRJ-CPF) from an alkali-solubilized fraction of RJ by C18 reverse-phase column chromatography. The elution profile by the C18 column chromatography and the molecular mass of the purified HBRJ-CPF material coincided with those of 10 hydroxy-2-decenoic acid (10H2DA). We then examined the collagen production promoting activities of several commercially available fatty acids contained in RJ. We found that 10H2DA and 10-hydroxydecanoic acid increased the collagen production in a dose-dependent manner. Furthermore, 10H2DA induced the fibroblast cell line, NHDF, to produce transforming growth factor-beta 1 (TGF-beta 1) which is an important factor for collagen production. As expected, the collagen production-promoting activity of 10H2DA was neutralized by the anti-TGF-beta 1 antibody. These result suggest that HBRJ-CPF identified as 10H2DA promoted the collagen production of AA-2G-treated fibroblasts by inducing TGF-beta 1 production. PMID- 15118302 TI - Characterization of sericin powder prepared from citric acid-degraded sericin polypeptides of the silkworm, Bombyx Mori. AB - Acid-degraded sericin powder (AC-SP) was prepared from aqueous solution containing citric acid-degraded sericin polypeptides of Bombyx mori. The morphological and biochemical properties of AC-SP were compared with those of alkali-degraded sericin powder (AL-SP) and hot-water degraded sericin powder (HW SP). Based on an SEM analysis, AC-SP showed a thin film structure of 10-100 microm with good dispersity while AL-SP and HW-SP had a much larger thin film structure (<500 microm). The extract of AC-SP showed stronger trypsin inhibitor activity due to cocoon shell trypsin inhibitor (CSTI-IV) than that of HW-SP. The extract of AL-SP showed no CSTI-IV activity. It was found that AC-SP was a trypsin inhibitor complex powder and that the release of CSTI-IV from AC-SP depended on pH and ion strength. Similar powder materials were obtained when such organic acids as tartaric acid and succinic acid were used. These results suggest that the acid-degraded sericin polypeptides work as a protein matrix to which CSTI-IV may bind ionically. PMID- 15118303 TI - Effects of a novel gaseous antioxidative system containing a rosemary extract on the oxidation induced by nitrogen dioxide and ultraviolet radiation. AB - Rosemary is commonly used as a spice and a flavoring agent in food processing. Although the antioxidative properties of its extracts have been investigated, there have been few reports on the volatile components of rosemary. We designed a novel antioxidative system which can generate the volatile constituents in the gaseous phase from a rosemary extract and evaluated the gaseous antioxidative activities against both lipid peroxidation and cell death induced by nitrogen dioxide and ultraviolet radiation. The antioxidative effects of the major volatile components on the oxidation of linoleic acid induced by azo compounds were also investigated in a solution. The volatile components in the novel antioxidative system suppressed the Jurkat cell death induced by nitrogen dioxide and the intracellular formation of reactive oxygen species in fibroblast cells induced by ultraviolet radiation. 1,8-Cineole among the volatile components exerted an antioxidative effect against the oxidation of linoleic acid in a solution induced by azo compounds and ultraviolet radiation. These data suggest that the volatile constituents of a rosemary extract had antioxidative properties and that gaseous exposure antioxidant is a promising method for promoting health. PMID- 15118304 TI - Multiplicity of 2,3-dihydroxybiphenyl dioxygenase genes in the Gram-positive polychlorinated biphenyl degrading bacterium Rhodococcus rhodochrous K37. AB - Rhodococcus rhodochrous K37, a Gram-positive bacterium grown under alkaline conditions, was isolated for its ability to metabolize PCBs. Analysis revealed that it has eight genes encoding extradiol dioxygenase, which has 2,3 dihydroxybiphenyl 1,2-dioxygenase activity, and these genes were designated bphC1 to bphC8. According to the classification of extradiol dioxygenases [Eltis, L. D., and Bolin, J. T., J. Bacteriol., 178, 5930-5937 (1996)], BphC3 and BphC6 belong to the type II enzyme group. The other six BphCs were classified as members of the type I extradiol dioxygenase group. BphC4 and BphC8 were classified into a new subfamily of type I, family 3. Two linear plasmids, 200 kb and 270 kb in size, were found in K37, and the bphC6 and bphC8 genes were located in the 200 kb linear plasmid. Northern hybridization analysis revealed that the bphC1, bphC2, and bphC7 genes were induced in the presence of testosterone, the bphC6 gene was induced by fluorene, and the bphC8 gene was induced by biphenyl. All eight BphC products exhibited much higher substrate activity for 2,3 dihydroxybiphenyl than for catechol, 3-methylcatechol, or 4-methylcatechol. PMID- 15118305 TI - Relationship between the preference for sake (Japanese rice wine) and the movements of metabolic parameters coinciding with sake intake. AB - Selection of foods by animals largely depends on their physiological condition. In this paper, we reported how physiological changes in rats after ingestion of sake (Japanese rice wine) affected their preference for different kinds of sake. Rats could discriminate among various kinds of sakes in a two-bottle choice test, even after adjustment of the glucose concentration or alcohol concentration of the sake, suggesting that the choice of a sake by animals was based on more than one ingredient. To identify effect of a rat's physiological condition on the selection of sake, we monitored the levels of blood glucose, nonesterified fatty acids (NEFA), and ketone bodies in serum in mice after forced intragastric ingestion of sake that had previously been offered to the rats in a two-bottle choice test. Blood glucose levels in mice were not different between the rats fed the palatable and unpalatable sakes, and the NEFA level and ketone level were high in rats fed the unpalatable sake. To further clarify the relationship between physiological condition and preference for sake, rats were offered eight kinds of Junmai-syu, which is made with only rice and no sub-ingredient. Rats could still discriminate among Junmai-syu. Furthermore, with two exceptions, the preference for a sake was significantly correlated with the ketone level. The order in the ratio of blood insulin level to blood glucagon level, which is an indicator of metabolism, was correlated with the order in preference to sake. These results suggest that some physiological factors besides oral stimulation also are important factors in the selection of a sake. PMID- 15118306 TI - Transient assay system for the analysis of PR-1a gene promoter in tobacco BY-2 cells. AB - In order to develop a rapid and versatile assay system suitable for the analysis of regulated expression of tobacco pathogenesis-related protein 1a (PR-1a) gene, we investigated the use of the transient gene expression system in tobacco BY-2 cells by microprojectile bombardment. Using dual luciferase assay as a reporter gene expression detection system, we observed significant induction of PR-1a promoter activity by salicylic acid (SA) treatment. On the other hand, treatment with 4-hydroxybenzoic acid (4-HBA) resulted in no detectable increase in luciferase activity. Co-expression of a trans-acting factor, the NPR1/NIM1 protein of Arabidopsis, resulted in the induction of higher expression levels of the PR-1a promoter. These results suggest that the assay system is applicable for the analysis of factors involved in the regulated expression of SA-inducible defense-related genes. PMID- 15118307 TI - Construction and characterization of chimeric proteins composed of type-1 and type-2 periplasmic binding proteins MglB and ArgT. AB - The respective type-1 and type-2 periplasmic binding proteins (PBPs) MglB and ArgT are believed to have evolved from a common ancestor into siblings showing topological differences in their main chain connectivity. At first glance, they show similar structure. But, more detailed examination reveals that the chain connectivity of ArgT is more convoluted than that of MglB. Reflecting that complexity, the folding of ArgT is complicated and involves intermediate folds. On the other hand, the folding of MglB is a simple two-state transition. In the present study, we constructed and characterized several chimeras made up of various subdomains of MglB and ArgT with the aim of gaining insight into the evolution of protein folding and protein structure. Although these chimeras did not fold as compactly as their parental proteins, some did exhibit cooperative folding, which suggests that novel proteins with new connectivity and new folding pathways could have emerged at a fairly high rate throughout the evolution of proteins. PMID- 15118308 TI - Alkali- and halo-tolerant catalase from Halomonas sp. SK1: overexpression in Escherichia coli, purification, characterization, and genetic modification. AB - A catalase gene, ohktA, from an alkali- and halo-tolerant bacterium, Halomonas sp. SK1, on the pKK223-3, was expressed in the catalase-lacking Escherichia coli strain UM2. Highly purified catalase showing a single band on SDS-PAGE was obtained by two liquid chromatography steps on DEAE-Toyopear1 and Chelating Sepharose Fast Flow. The enzyme, oHktA, shows high catalase activity with a pH optimum at 10, and the activity was stable in 4 M KC1. This enzyme is thermo sensitive, showing a significant loss of activity within 5 minutes at 37 degrees C. To modify the stability of the catalase, the addition of domain II of the heat stable Mn catalase from Thermus thermophilus to the C-terminus was made. When coexpressed with a chaperone (PhFKBP29) gene product, peptidyl-prolyl cis-trans isomerase, from a thermophilic bacterium, a chimeric catalase was produced in the soluble fraction. The stability of this catalase in the range of 37 degrees -45 degrees C was improved and it was stable for more than 1 h at 37 degrees C. PMID- 15118309 TI - New brasiliamide congeners, brasiliamides C, D and E, from Penicillium brasilianum Batista JV-379. AB - Three new brasiliamide congeners, brasiliamides C, D and E, were isolated from okara fermented with Penicillium brasilianum Batista JV-379. Their structures were elucidated on the basis of spectral data and chemical evidence. NMR spectra of these brasiliamides exhibited a mixture of four or two conformers due to the restricted rotation of an amide bond in a solution. The (1)H- and (13)C-NMR spectral data were analyzed for a major rotamer at an appropriate temperature, since the signals were broadened at room temperature. Both brasiliamides C and D showed convulsive activity against silkworms with an ED(50) value of 400 microg/g of diet, whereas brasiliamide E showed less activity than the others. PMID- 15118310 TI - Effects of physical fatigue in mice on learning performance in a water maze. AB - We investigated the effects of physical fatigue produced by swimming exercise on learning the Morris water maze in BALB/c mice. We measured the escape latency in the maze immediately after the swimming exercise. The control group was soaked in the water but not fatigued. For easier tasks, like one with an obvious cue flag, the escape latency was not changed by exercise fatigue. However, escape latency was increased after exercise fatigue for more difficult tasks of spatial learning. These results appear to suggest that physical fatigue impaired learning performance. The effects of swimming exercise fatigue on learning efficiency were then investigated. Mice were continuously fatigued during the spatial learning period. This increased escape latency between the first and third sessions. The results suggest that learning efficiency was impaired by exercise fatigue. This system may be useful for screening new foods used to enhance brain function during exercise. PMID- 15118311 TI - In vivo and in vitro interactions of the Bombyx mori chymotrypsin inhibitor b1 with Escherichia coli. AB - Various chymotrypsin inhibitors occur in the hemolymph of silkworm larvae. Interaction of chymotrypsin inhibitor b1 (CI-b1) with Escherichia coli was examined from the viewpoint of action against invading bacteria. Injection of dead E. coli cells into larva reduced the CI-b1 content of the hemolymph, suggesting in vivo binding of CI-b1 to the outer membrane of the cell. Results from incubation of E. coli in cell-free hemolymph in the presence or absence of lipopolysaccharide indicated that CI-b1 is the only CI bound to E. coli and that it interacts with lipopolysaccharide. CI-b1 formed a complex with lipopolysaccharide in vitro; the value of the dissociation constant was relatively large. Inhibitory activity of CI-b1 changed insignificantly in mixture with lipopolysaccharide. CI-b1 affected the growth of E. coli but never worked lethally. CI-b1 is speculated to be a mediator that scavenges intruding bacteria rather than a direct anti-bacterial factor. This is the first report confirming that CI-b1 is a lipopolysaccharide binding protein. PMID- 15118312 TI - Involvement of tyrosines at fucose-binding sites of Aleuria aurantia lectin: non equal response to site-directed mutagenesis among five sites. AB - Since the involvement of Tyr residues in the fucose-binding of Aleuria aurantia lectin (AAL) was proved by chemical modification using the Tyr-specific reagent tetranitromethane, site-directed mutagenesis was attempted. Since the tertiary structure of AAL was determined recently to be a six-bladed beta-propeller fold, and five fucose-binding sites per subunit were found, based on positions of Tyr residues in the tertiary structure, three classes of mutants were constructed: 1) Tyr on the 2nd beta-strand of each blade (beta-2 mutants), 2) Tyr or Trp on the 3rd beta-strand (beta-3 mutants), and 3) Tyr outside of binding sites (other-Y mutants). The mutagenized cDNA was expressed in Escherichia coli as His-tag-AAL, and the hemagglutinating activity was assayed. Among 14 mutants, three beta-2 mutants (Y26A, Y79A, and Y181A), and three beta-3 mutants (Y92A, W149A, and Y241A) showed decreased activity. These mutated residues resided at Sites 1, 2, and 4, at the same locations relatively in the binding sites. Mutagenesis of Tyr or Trp at the corresponding locations in Sites 3 and 5 did not lead to a reduction in activity. Results indicate that the properties of Sites 1, 2, and 4 are different from those of Sites 3 and 5, and that the contribution of these two sites to the hemagglutination reaction was minor. PMID- 15118313 TI - Zooxanthellactone, a novel gamma-lactone-type oxylipine from dinoflagellates of Symbiodinium sp.: structure, distribution, and biological activity. AB - A novel fatty acid derivative named zooxanthellactone (ZL) was isolated from several strains of symbiotic microalgae, dinoflagellates of the genus Symbiodinium. The metabolite is structurally related to docosahexaenoic acid (DHA) and seems to be biosynthesized by oxidation and subsequent lactonization. The absolute stereochemistry was determined from the specific rotation of the perhydro derivative. The distribution of ZL within several Symbiodinium isolates was quantitatively analyzed by HPLC techniques and suggested a relationship between the productivity of this metabolite and the Symbiodinium phylogeny. The cytotoxicity of ZL was evaluated by using human squamous cell carcinoma cell lines in comparison with that of DHA and other common fatty acids, suggesting that the long unsaturated chain was important rather than the gamma-lactone moiety. PMID- 15118314 TI - No detectable transfer of dietary lactoferrin or its functional fragments to portal blood in healthy adult rats. AB - We investigated the transfer of dietary bovine lactoferrin (LF) and its functional lactoferricin (LFcin) B-containing fragments to the portal blood of healthy adult rats by using several techniques. After a single administration of (125)I-labeled LF, radioactive bands were detected in autoradioluminograms of the portal blood, but similar bands were also observed after the administration of [(125)I]NaI. Although ovalbumin was detected by ELISA at 3-18 ng/ml in the portal blood plasma after an overnight administration, no LF was detected (< or =1.5 ng/ml). The antibody-captured ovalbumin fragments, but not the LF fragments, were detected in the plasma by surface-enhanced laser desorption/ionization affinity mass spectrometry (SELDI affinity MS). We finally attempted to detect the LFcin B containing fragments by SELDI affinity MS with on-chip LFcin B-conversion, but could not detect them (< or =1 ng/ml) in the portal blood after the LF ingestion. The level of LF or its functional fragments transferred to the portal blood was therefore extremely low, if any. PMID- 15118315 TI - Preparation and structural analysis of actinidain-processed atelocollagen of yellowfin tuna (Thunnus albacares). AB - Pepsin-hydrolyzed collagen (atelocollagen) is a trimer, consisting of alpha 1 and alpha 2 monomers, and shows molecular species corresponding to a monomer, dimer (beta chain), and trimer (gamma chain) by SDS-polyacrylamide gel electrophoresis. Atelocollagen was purified from yellowfin tuna (Thunnus albacares) by salt precipitation and cation-exchange chromatography. Enzymatic hydrolysis of the atelocollagen by actinidain, a cysteine protease purified from kiwifruit, was analyzed by SDS-polyacrylamide gel electrophoresis. The triple helical structure unique to collagen was retained in the atelocollagen as judged by circular dichroism spectra. The actinidain-processed atelocollagen showed only monomeric alpha 1 and alpha 2 chains, with no beta and gamma chains, by SDS-polyacrylamide gel electrophoresis; nevertheless, it retained the typical triple helical structure. It is suggested that actinidain cleaved the atelocollagen molecule at specific sites on the inside of the inter-strand cross-linking peptides. PMID- 15118316 TI - Fractionation and anti-tumor activity of the mycelia of liquid-cultured Phellinus linteus. AB - All the fractions of Phellinus linteus mycelia showed anti-tumor activity toward solid tumors planted in mice. The highest anti-tumor activity of 81.2% was observed in the protein-glucan complex obtained by precipitating the 24% NaOH extract at pH 6.0. This protein-glucan complex consisted of 39.3% polysaccharide and 49.4% protein. Its (13)C- and (1)H-NMR data showed that the main glucan part of the complex was simple alpha-1,3-glucan chains. PMID- 15118317 TI - Enhanced resistance to blast fungus and bacterial blight in transgenic rice constitutively expressing OsSBP, a rice homologue of mammalian selenium-binding proteins. AB - The rice Oryza sativa selenium-binding protein homologue (OsSBP) gene encodes a homologue of mammalian selenium-binding proteins, and it has been isolated as one of the genes induced by treating a plant with a cerebroside elicitor from rice blast fungus. The possible role of OsSBP in plant defense was evaluated by using a transgenic approach. Plants overexpressing OsSBP showed enhanced resistance to a virulent strain of rice blast fungus as well as to rice bacterial blight. The expression of defense-related genes and the accumulation of phytoalexin after infection by rice blast fungus were accelerated in the OsSBP overexpressors. A higher level of H(2)O(2) accumulation and reduced activity of such scavenging enzymes as ascorbate peroxidase and catalase were seen when the OsSBP overexpressing plants were treated with the protein phosphatase 1 inhibitor, calyculin A. These results suggest that the upregulation of OsSBP expression conferred enhanced tolerance to different pathogens, possibly by increasing plant sensitivity to endogenous defense responses. Additionally, the OsSBP protein might have a role in modulating the defense mechanism to biotic stress in rice. PMID- 15118318 TI - Immuno-potentiating effects of the antler-shaped fruiting body of Ganoderma lucidum (Rokkaku-Reishi). AB - The immuno-potentiating effects of the antler-shaped fruiting body of Ganoderma lucidum (Rokkaku-Reishi, RR), which has been used as a traditional supplement for human health, were investigated in mice. BALB/c mice were administered orally with RR for 3 days at a dose of 50 mg/kg or 500 mg/kg, and interferon-gamma (IFN gamma) production by splenocytes in response to lipopolysaccharide (LPS) was examined on day 4. The oral administration of 500 mg/kg of RR resulted in a significant increase (p<0.05) in IFN-gamma production. Stimulation of splenic adherent cells from these mice with LPS also resulted in a significant increase (p<0.05) in interleukin-12 (IL-12) production compared with that from the control mice, suggesting that splenic macrophages were activated by RR administration. Furthermore, 500 mg/kg of RR administered for 14 days resulted in a significant increase (p<0.05) in IFN-gamma production by splenocytes in response to both LPS and concanavalin A (Con A). These results suggest that not only splenic macrophages but also T cells were activated by the long-term treatment with RR in vivo. On the other hand, the production of interleukin-4 (IL-4), which is known as an allergic disease-related cytokine, was not affected by the long-term treatment with RR. Our results suggest that the oral administration of RR resulted in Th1-associated immuno-potentiating activities in vivo. PMID- 15118319 TI - Inverse correlation between the nitrogen balance and induction of rat liver serine dehydratase (SDH) by dietary protein. AB - Rats of different ages (3 to 15-wk-old) were fed on a 25% casein diet for one week, and the nitrogen balance and liver serine dehydratase (SDH, EC 4.2.1.13) activity were then determined. The value for nitrogen balance decreased with the age of the rats, while the liver SDH activity increased. A statistical analysis showed clear inverse correlation between the two factors (R(2) = 0.7372, p < 0.01). This result suggests that SDH was induced by response to the amount of surplus amino acids from dietary protein taken beyond the body's requirement. The increase in SDH activity was accompanied by an increase in the level of SDH mRNA. Since the half-life of this mRNA did not change significantly, the induction was mainly controlled at the level of transcription. In addition, the induction seems not to be related to gluconeogenesis, since the mRNA levels of tyrosine aminotransferase (TAT) and phosphoenolpyruvate carboxykinase (PEPCK), other gluconeogenic enzymes, were not changed under these experimental conditions. PMID- 15118320 TI - Characterization of murine grancalcin specifically expressed in leukocytes and its possible role in host defense against bacterial infection. AB - Such phagocytic leukocytes as macrophages and neutrophils are the key cellular components of innate immunity. The actin cytoskeleton is essential for their recruitment and activation in infected tissues. We have previously identified p65/L-plastin with Ca(2+)-, calmodulin-, and beta-actin-binding domains in macrophages. In order to further investigate the p65/L-plastin-involved cellular functions, we cloned the cDNA for murine grancalcin, a possible binding partner of p65/L-plastin. According to the sequence, grancalcin is a member of the penta EF-hand protein family. We prepared recombinant (r) grancalcin for functional studies and found that it exhibited Ca(2+)-dependent precipitation. High-titer antibodies against the protein enabled us to detect intracellular grancalcin. A flow cytometric analysis revealed grancalcin to be highly expressed in macrophages and neutrophils. The protein was particularly abundant in those cells recovered from bacteria-infected sites. Immunohistochemical studies clarified that grancalcin was translocated to the actin cytoskeleton in macrophages upon exposure to bacterial lipopolysaccharide. These findings suggest that grancalcin plays a key role in leukocyte-specific functions that are responsible for host defense. PMID- 15118321 TI - Novel lipoxygenase inhibitors, tetrapetalone B, C, and D from Streptomyces sp. AB - Three novel lipoxygenase inhibitors, tetrapetalone B (2, C(28)H(35)NO(9)), C (3, C(26)H(34)NO(8)), and D (4, C(28)H(36)NO(10)), were isolated from a culture broth of Streptomyces sp. USF-4727 that produced a lipoxygenase inhibitor tetrapetalone A (1) simultaneously. Each chemical structure was revealed by spectroscopic evidence, this suggests that these three compounds are structurally related to 1. They had a tetracyclic skeleton and a beta-D-rhodinosyl moiety. Tetrapetalone B, C, and D inhibited soybean lipoxygenase with IC(50): 320, 360, and 340 microM respectively. PMID- 15118323 TI - Simple determination of trace amounts of anionic surfactants in river water by spectrophotometry combined with solid-phase extraction. AB - A simple and sensitive spectrophotometric method combined with solid-phase extraction (SPE) for the simultaneous determination of sodium linear dodecylbenzenesulfonate (DBS) and sodium dodecyl sulfate (SDS) is described. The C2 (ethyl group bonded silicagel) cartridge could be repeatedly used more than 500 times for SPE, and it enabled the anionic surfactants to be concentrated by 50-fold. The calibration graph for DBS was linear in the range from 1.6 x 10(-8) M to 5.0 x 10(-7) M and for SDS from 2.0 x 10(-9) M to 3.0 x 10(-7) M. The relative standard deviation (n=5) for 5.0 x 10(-7) M DBS was 3.1% and for 2.5 x 10(-7) M SDS was 1.7%. The proposed method was applied to the simultaneous determination of DBS and SDS in river-water samples. PMID- 15118322 TI - RSC Nucleosome-remodeling complex plays prominent roles in transcriptional regulation throughout budding yeast gametogenesis. AB - RSC is a nucleosome-remodeling complex of Saccharomyces cerevisiae essential for growth that can alter histone-DNA interaction by using the energy of ATP hydrolysis. Nps1p/Sth1p is an ATPase subunit of RSC. A mutation in the conserved ATPase domain of Nps1p causes a sporulation defect with decreased expression of early meiotic genes, especially IME2. This defect is partially suppressed by the overexpression of either IME1 or IME2. A homozygous diploid of a novel temperature-sensitive nps1 mutation, nps1-13, harboring amino acid substitutions within the bromodomain, was unable to sporulate. Overexpression of IME, IME2, or both of these genes allowed the completion of meiosis I and meiosis II in nps1-13 but not the formation of mature asci. In nps1-13 carrying YEpIME1, the expression of a group of sporulation-specific genes, which express at the middle stages of sporulation and are required for spore-wall formation, notably diminished, and several late sporulation genes expressed at the early stages of sporulation. These results suggest that Nps1p/RSC plays important roles during the spore development process by controlling gene expression for initiating both meiosis and spore morphogenesis, and ensures proper expression timing of late meiotic genes. PMID- 15118324 TI - Interaction between a type-II dockerin domain and a type-II cohesin domain from Clostridium thermocellum cellulosome. AB - The interaction between the type-II dockerin domain of the scaffoldin protein CipA and the type-II cohesin domain of the outer layer protein SdbA is the fundamental mechanism for anchoring the cellulosome to the cell surface of Clostridium thermocellum. We constructed and purified a dockerin polypeptide and a cohesin polypeptide, and determined affinity constants of the interaction between them by the surface plasmon resonance method. The dissociation constant (K(D)) value was 1.8 x 10(-9) M, which is a little larger than that for the combination of a type-I dockerin and a type-I cohesin. PMID- 15118325 TI - Effect of the rate of temperature increase on water quality during heating in electromagnetic- and gas-heated pans. AB - More rapid increases in the pH value and hardness during electromagnetic heating of a pan of water were observed than when the pan was heated by LNG or LPG. The water quality changed universally in several tap water samples across Japan. This quality change was closely correlated with the rate of temperature increase, irrespective of heating by electromagnetic induction, LNG or LPG. PMID- 15118326 TI - Heterologous mevalonate production in Streptomyces lividans TK23. AB - Mevalonate is a ubiquitous biosynthetic intermediate of terpenoids and is used as a moisturizer in cosmetics and a chemical for biochemical research. In this study, we have achieved a heterologous production of this useful compound by expression in Streptomyces lividans TK23 of 3-hydroxy-3-methylglutaryl-CoA synthase and 3-hydroxy-3-methylglutaryl-CoA reductase genes, which were cloned from Streptomyces sp. strain CL190. PMID- 15118327 TI - Suppression of dioxin mediated aryl hydrocarbon receptor transformation by ethanolic extracts of propolis. AB - Present study demonstrated that the ethanolic extracts of propolis containing higher concentrations of flavonoids suppressed 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD)-induced aryl hydrocarbon receptor transformation in a dose dependent manner. The IC(50) values of propolis group 3 and group 12 were 1.2 and 3.6 microg/ml, respectively, indicating that propolis showed stronger antagonistic effects as compared with vegetable extracts. PMID- 15118328 TI - In vivo bioconversion of tetrahydroisoquinoline by recombinant coclaurine N methyltransferase. AB - Coclaurine N-methyltransferase from Coptis japonica catalyzes the N-methylation of coclaurine as well as simple tetrahydroisoquinoline. We examined the possibility of converting 6,7-dimethoxy-1,2,3,4-tetrahydroisoquinoline into its N methylated product using transgenic Escherichia coli, which expressed recombinant coclaurine N-methyltransferase, without the addition of a methyl-group donor. Transgenic E. coli successfully N-methylated the substrate added to the medium and excreted the product. Limitation of bioconversion by the supply of methyl group donor is discussed. PMID- 15118329 TI - A new bipyrrole and some phenolic constituents in prunes (Prunus domestica L.) and their oxygen radical absorbance capacity (ORAC). AB - Isolation and structural elucidation of prune constituents were performed and total 10 compounds were determined by NMR and MS analyses. A novel compound was identified to be 2-(5-hydroxymethyl-2',5'-dioxo-2',3',4',5'-tetrahydro-1'H-1,3' bipyrrole)carbaldehyde, and 7 phenolic compounds were isolated from prunes for the first time. In addition, antioxidant activity of them was evaluated on the basis of the oxygen radical absorbance capacity (ORAC). PMID- 15118330 TI - Substrate specificity of aminopeptidase from the mid-gut gland of the scallop (Patinopecten yessoensis). AB - An action for various peptides and a kinetic study for amino acid p-nitroanilides (pNAs) and 4-methylcoumaryl-7-amides (MCAs) were performed with purified aminopeptidase from the mid-gut of the scallop. The enzyme preferred dipeptides having Ala, Met, and Phe in the amino-terminal or the penultimate position from the amino-termini. The catalytic efficiencies, k(cat)/K(m) values for Ala-pNA and MCA were the highest in the tested substrates, and those for pNA and MCA substrates having Met or Phe were the next highest. The enzyme was found to be a new alanine-specific aminopeptidase. PMID- 15118331 TI - Improved expression of novel red- and green-emitting luciferases of Phrixothrix railroad worms in mammalian cells. AB - Luciferases are widely used for the quantitative monitoring of gene expression in a variety of organisms. We successfully expressed novel red- and green-emitting luciferases of Phrixothrix railroad worms in mammalian cells in combination with the Kozak sequence and the CAG promoter. The characteristic properties of these luciferases indicate that they are appropriate reporter genes for the simultaneous monitoring of two gene expressions. PMID- 15118332 TI - Brassinosteroid selectively regulates PIN gene expression in Arabidopsis. AB - When brassinosteroid (BR)-deficient mutant (det2) or wild-type (WT) seedlings were treated with brassinolide (BL), the most active BR, for 3 h, the abundance of PIN4 and PIN7 transcripts decreased, and there were fewer PIN4 and PIN7 transcripts in det2 than in the WT. This suggests that BL selectively regulates the PIN gene in a complex manner. PMID- 15118333 TI - Zooxanthellamide B, a novel large polyhydroxy metabolite from a marine dinoflagellate of Symbiodinium sp. AB - Zooxanthellamide B, C(128)H(220)N(2)O(53)S(2), a polyhydroxy secondary metabolite, was isolated from a cultured marine dinoflagellate of the genus Symbiodinium. A detailed 2D NMR analysis revealed the chemical structure as a delta-lactone analogue of zooxanthellamide A, which had previously been isolated from the same dinoflagellate by us. The relative configuration of the delta lactone moiety was determined by NOE experiments and a coupling constant analysis, and that of other ring systems was found to be the same as zooxanthellamide A by the chemical correlation between zooxanthellamides A and B. PMID- 15118335 TI - Thr/Ser-rich domain of Aspergillus glucoamylase is essential for secretion. AB - The recombinant Aspergillus awamori strain carrying the mutant glucoamylase encoding gene in which the entire Thr/Ser-rich Gp-I domain was deleted abolished secretion of mutant glucoamylase. The transcription of the Bip-encoding bipA was low in the wild type (wt) strain, but elevated in the recombinant strain under the condition of glaA expression. The results indicate that the Gp-I domain is vital for glucoamylase secretion. PMID- 15118334 TI - Properties of mycelial aggregate-specific lectin of Pleurotus cornucopiae produced in Pichia pastoris. AB - cDNA of a mycelial aggregate-specific lectin of Pleurotus cornucopiae was expressed in Pichia pastoris, and the expression product was purified and characterized. The product was functional, and the hemagglutinating activity was inhibited most strongly by the addition of N-acetyl-D-galactosamine as was the native lectin. The native lectin is a glycoprotein having five glycosylation recognition signals, and the expression product showed slightly larger molecular mass than that of the native one due to further glycosylation. PMID- 15118336 TI - 3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]indole induces apoptosis and necrosis with activation of different caspases in rat splenocytes. AB - A dietary carcinogen, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) at 20 microM activates caspase-3-like proteases as an apoptotic marker in rat splenocytes. The present study demonstrated 100 microM Trp-P-1 induced necrosis with activation of caspase-3-like proteases. The activation in necrosis and apoptosis resulted from the activation of caspase-9 and caspase-8, respectively. Thus, Trp-P-1 induces apoptosis and necrosis with the activation of different caspases. PMID- 15118337 TI - Effect of ethanol on cell growth of budding yeast: genes that are important for cell growth in the presence of ethanol. AB - The budding yeast Saccharomyces cerevisiae has been used in the fermentation of various kinds of alcoholic beverages. But the effect of ethanol on the cell growth of this yeast is poorly understood. This study shows that the addition of ethanol causes a cell-cycle delay associated with a transient dispersion of F actin cytoskeleton, resulting in an increase in cell size. We found that the tyrosine kinase Swe1, the negative regulator of Cdc28-Clb kinase, is related to the regulation of cell growth in the presence of ethanol. Indeed, the increase in cell size due to ethanol was partially abolished in the SWE1-deleted cells, and the amount of Swe1 protein increased transiently in the presence of ethanol. These results indicated that Swe1 is involved in cell size control in the presence of ethanol, and that a signal produced by ethanol causes a transient up regulation of Swe1. Further we investigated comprehensively the ethanol-sensitive strains in the complete set of 4847 non-essential gene deletions and identified at least 256 genes that are important for cell growth in the presence of ethanol. PMID- 15118338 TI - Differential expression of three plastidial sigma factors, OsSIG1, OsSIG2A, and OsSIG2B, during leaf development in rice. AB - We isolated and characterized two rice nuclear genes, OsSIG2A and OsSIG2B, encoding the putative sigma-factor of the plastid RNA polymerase. Deduced protein sequences predicted a plastid-localizing signal in the N-terminus and subsequent polypeptides similar to known SIG2 proteins. Gene expression analysis revealed that the OsSIG2A transcript is more abundant than the OsSIG2B transcript in all tissues tested and that both rice SIG2s are expressed from earlier stages of leaf development than that in the case of OsSIG1. These results indicate differential expression of SIG genes in leaf morphogenesis, suggesting the existence of tissue and stage-specific functions of SIG proteins for transcriptional regulation of chloroplast genes in plant development. PMID- 15118339 TI - Unusual transcription regulation of the niaD gene under anaerobic conditions supporting fungal ammonia fermentation. AB - The niaD gene of the fungus Aspergillus nidulans encodes an assimilatory nitrate reductase and exogenous ammonium represses its expression. Under anoxic conditions, however, A. nidulans expressed niaD even in the presence of ammonium and used the gene product for dissimilatory nitrate reduction (ammonia fermentation). This transcription regulation mechanism under anaerobiosis is critical for the fungus to ferment ammonium. PMID- 15118340 TI - Pulse pressure and left ventricular diastolic dysfunction in hypertensive patients. AB - BACKGROUND: Impaired left ventricular diastolic function is a common finding in essential hypertension. METHODS: In order to investigate possible relationships between flow velocity through the mitral valve (E/A; index of left ventricular diastolic function) and 24-hour blood pressure and heart rate variations, 198 patients with mild to moderate essential hypertension were studied by Doppler echocardiography and ambulatory blood pressure monitoring. They were divided according to age into group 1 (n = 88, age 40-54 years) and group 2 (n = 110, age 55-79 years). Each group was divided into subgroups with (1a, 2a) or without (1b, 2b) left ventricular hypertrophy according to the end-diastolic posterior wall thickness and/or the interventricular septum thickness. RESULTS: In a multivariate stepwise regression analysis, age (beta = -0.25, p < 0.0001), posterior wall thickness (beta = -0.31, p < 0.0057) and mean heart rate during the day (beta = -0.34, p < 0.0284) were the independent predictors of E/A in the pooled population. In group 1a (young subjects with left ventricular hypertrophy), mean systolic blood pressure during the night (beta = -0.33, p < 0.041) was the only independent predictor of E/A. In the elderly group without left ventricular hypertrophy (group 2b), the mean heart rate during the day (beta = -0.44, p < 0.0000) and mean pulse pressure during the night (beta = -0.60, p < 0.0007) were the independent predictors of E/A. CONCLUSIONS: The new finding provided by this study is that in elderly hypertensive patients without left ventricular hypertrophy, a large pulse pressure at night may serve as an independent predictor of abnormal left ventricular diastolic filling. PMID- 15118341 TI - Hypoplasia of the parabrachial/kolliker-fuse complex in perinatal death. AB - We report the first observation of perinatal death attributable to morphological alterations of the parabrachial/Kolliker-Fuse complex. In an infant, born at 41 weeks of gestation with severe signs of asphyxia and dead 20 h after delivery, we observed the presence of rare and immature neurons in the brain stem areas of both lateral and medial parabrachial nuclei and the absence of the characteristic neurons of the Kolliker-Fuse nucleus. Such hypoplasia was associated with severe hypoplasia of the arcuate nucleus and pulmonary hypoplasia. PMID- 15118342 TI - Transferrin microheterogeneity in fetal blood. AB - OBJECTIVES: To investigate the distribution of microheterogeneous subfractions of transferrin in fetal blood and the influence of highly sialylated transferrins on fetal growth. STUDY METHOD: Serum transferrin concentrations were determined by a standard turbidimetric assay. Microheterogeneous transferrin subfractions were assessed by crossed immunoisoelectric focusing. RESULTS: In normal term infants, total serum transferrin concentrations and percent distribution of highly sialylated transferrins (> or = 5-sialo-transferrins) were markedly lower; the percent distributions of hyposialylated transferrins (0- and 1-sialo transferrins) were apparently higher than those in non-pregnant and pregnant women. There was no significant positive correlation between the serum concentrations of total transferrin or highly sialylated transferrins in infants' blood and birth weights (r = 0.187, p = 0.582; r = 0.374, p = 0.257, respectively). CONCLUSION: The transferrin microheterogeneity pattern shifted towards reduced glycosylation and sialylation in addition to a decrease in total transferrin concentration in fetal blood compared to that of non-pregnant and pregnant women. The concentrations of serum total transferrin and the highly sialylated transferrins in fetal blood, if higher than a certain level, did not seem to have any influence on normal fetal growth. PMID- 15118343 TI - Pretransplant mixed lymphocyte culture still has an impact on graft survival. AB - BACKGROUND/AIM: Mixed lymphocyte culture (MLC) is an important in vitro test for studying allograft reaction. The recipient-donor MLC reflects donor specific hypo or hyperimmune response. The various studies have been correlated donor-specific MLC reactivity with graft survival in cadaver kidney transplantation. This retrospective study reports the relationship between of MLC hyper-responsiveness and graft survival in living-donor kidney transplantation. METHODS: The study included 477 patients who underwent live-donor kidney transplantation between Marsh 1976 and January 2002. They were divided according to the relative response (RR) of pre transplant one way MLC into hypo responders (RR <4) and hyper responders (RR >4). The demographic and follow-up data for both groups were recorded. The duration of follow-up ranged from 44 to 84 months. RESULTS: The two groups were homogeneous regarding age, sex, donor source, HLA-A, B-and -DR mismatches, number of blood transfusion and type of 3 primary immunosuppressions. Acute rejection (AR) episodes were seen in 182 (44.8%) patients in the hypo responders group, of whom 10 were steroid resistant, while in the hyper-responder recipients, AR episodes occurred in 33 patients (46.5%) of whom 5 patients (15.2%) were steroid resistant (p < 0.05). Chronic allograft nephropathy occurred in 14.5% and 27.3% and the actuarial 5-year graft survival was 79% and 60% (p = 0.03).in the hypo- and hyper-responder groups, respectively. CONCLUSIONS: It is concluded that MLC reaction may predict high-risk patients for immunological graft failure and that more potent immunosuppression should be considered in MLC hyper-responders. PMID- 15118344 TI - Induction of manganese-superoxide dismutase by YS 51, a synthetic 1-(beta naphtylmethyl)6,7-dihydroxy- 1,2,3,4-tetrahydroisoquinoline alkaloid: implication for anti-inflammatory actions. AB - The effect of YS 51, a synthetic 1-(beta-naphtylmethyl)6,7-dihydroxy-1,2,3,4 tetrahydroisoquinoline alkaloid, on the expression of manganese-superoxide dismutase (Mn-SOD), an antioxidant enzyme, was examined in sheep pulmonary artery endothelial cells (SPAEC) and a human cervical carcinoma cell line (Hela). YS 51 alone or in combination with cytokines enhanced the expression of Mn-SOD mRNA in SPAEC and Hela cells. YS 51 also showed synergistic effects on the induction of Mn-SOD mRNA with phorbol-12-myristate-13-acetate (TPA) and/or tumor necrosis factor-alpha (TNF-alpha). In Hela cells, the induction of Mn-SOD mRNA by YS 51 was in a time- and dose-dependent manner and the expression of Mn-SOD mRNA was increased to a maximum of 4-fold in 9 h. Enhancement of Mn-SOD mRNA by YS 51 was completely abolished by actinomycin D but not cycloheximide, suggesting that the induction of Mn-SOD mRNA byYS 51 is independent of new protein synthesis. Pretreatment of curcumin, an inhibitor of c-jun N-terminal kinase (JNK), dose dependently suppressed the induction of Mn-SOD mRNA by YS 51, but not by 2'-amino 3'-methoxyflavone (PD98059) and 4-(4-fluorophenyl)-2-(4-methylsulfonylphenyl)-5 (4-pyridyl)imidazol (SB203580), inhibitors of mitogen-activated protein kinase. Also, YS 51 induced the phosphorylation activity of JNK in a time-dependent manner without affecting the phosphorylation activity of the extracellular signal regulated kinase 1 (ERK1) and p38 MAP kinase. These results implicated that the JNK pathway appears to play a crucial role in mediating the YS 51-induced Mn-SOD gene expression, and that up-regulation of Mn-SOD would contribute to the anti inflammatory actions mediated by YS 51. PMID- 15118345 TI - Vasoconstrictor responses to 5-hydroxytryptamine in the autoperfused hindquarters of spontaneously hypertensive rats. AB - In this work we studied the responses and receptors involved in the effects of intra-arterial 5-hydroxytryptamine (5-HT) in the in situ autoperfused hindquarters of spontaneously hypertensive rats (SHR). Intra-arterial administration of the highest doses (50-1,000 ng/kg) produced a vasoconstrictor effect that was inhibited by ritanserin (a selective 5-HT2 receptor antagonist), SB 206553 (a selective 5-HT(2B/2C) receptor antagonist) and spiperone (a nonspecific 5-HT(1/2A) receptor antagonist), and was mimicked by alpha-methyl-5 HT (a selective 5-HT2 receptor agonist) and m-CPP (a selective 5-HT2C receptor agonist), but not by the intra-arterial administration of BW 723C86, a selective 5HT2B receptor agonist. SB 206553 and spiperone inhibited alpha-methyl-5HT induced vasoconstriction in the hindquarters of SHR. Our data suggest that the vasoconstrictor response induced by 5-HT in the autoperfused hindquarters of SHR is mainly mediated by the activation of 5-HT2A and 5-HT2C receptors. PMID- 15118346 TI - Increased tracheal responsiveness to beta-adrenergic agonist and antagonist in ovalbumin-sensitized guinea pigs. AB - Despite the controversy of bronchial responsiveness to beta2-agonist drugs in asthma, in a previous study we have shown increased responsiveness of asthmatic tracheobronchial tree to isoprenaline. Therefore, in the present study, tracheal responsiveness to isoprenaline and also beta-adrenergic receptor blockade were studied in sensitized guinea pigs. An experimental model of asthma was induced in guinea pigs by sensitization of animals with injection and inhalation of ovalbumin (OA). The responses of tracheal chains of sensitized and control animals to cumulative concentrations of isoprenaline (I) in the absence and presence of 10 nmol/l propranolol were measured, and the effective concentration of I causing 50% of maximum response (EC50 I) was obtained. The propranolol blockade (CR - 1) was calculated by: (post-propranolol EC50 I/EC50 I) - 1. Tracheal responses of sensitized and control animals to cumulative concentrations of methacholine (M) were also measured and EC50 M were obtained. The tracheal responses of sensitized guinea pig to isoprenaline was significantly higher than that of the control animals (EC50 I for sensitized and control animals were 0.24 +/- 0.04 and 0.58 +/- 0.07 micromol/l, respectively; p < 0.001). The beta adrenergic receptor blockade by propranolol (CR - 1) was also significantly higher in sensitized guinea pigs than that of the control animals (p < 0.001). The results of this study indicate an increased tracheal response to beta adrenergic-stimulating drug and enhancement of beta-adrenergic blockade by propranolol in the sensitized guinea pig. PMID- 15118347 TI - Protein kinase C regulates alpha(2A/D)-adrenoceptor constitutive activity. AB - We investigated a possible role for protein kinases in the constitutive activity of alpha(2A/D) adrenoceptors in membranes from transfected PC12 cells, using a [35S]GTPgammaS binding assay. After treatment of intact cells with various protein kinase inhibitors, constitutive activity was assessed by the reduction in basal GTP binding caused by the inverse agonist rauwolscine (RAU). Inhibitors of protein kinase C (PKC) caused the loss of RAU-sensitive GTP binding, while inhibitors of other protein kinases were ineffective. Anti-G(alpha) antibody treatments showed that constitutive alpha(2A/D)-receptor activity is directed toward different G proteins than agonist-stimulated activity. T373A mutant receptors exhibited increased constitutive activity, including a component that was insensitive to PKC inhibition. Since T373 is located within a putative G(i/o) activator sequence, these results suggest that PKC-dependent phosphorylation of T373 increases alpha(2A/D)-adrenergic receptor constitutive activity and causes a switch in G protein preference. PMID- 15118348 TI - Hypotensive effects of eugenosedin-A with serotonin, alpha- and beta-adrenoceptor antagonistic activities in spontaneously hypertensive and normotensive rats. AB - Eugenosedin-A is a newly synthesized compound with special serotonergic, alpha- and beta1-adrenergic blocking actions. Intravenous injection of eugenosedin-A significantly caused dose-dependent decreases in the mean arterial blood pressure and heart rate in normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR). The effects of eugenosedin-A-decreased blood pressure and heart rate in SHR were more potent than in WKY. In in vitro experiments, eugenosedin-A competitively antagonized the serotonin-, norepinephrine- and clonidine-induced vasocontraction in a concentration-dependent manner in isolated thoracic aorta of WKY and SHR. We also observed that eugenosedin-A competitively antagonized the isoproterenol-induced positive inotropic effects in a concentration-dependent manner in the isolated left atrium of WKY and SHR. These findings clearly suggested that eugenosedin-A possesses alpha1/alpha2, beta1 and 5-HT2A receptor blocking activities. The order of pA2 values in isolated tissues of WKY was 5 HT2A > alpha1/alpha2 > beta1. However, the order of pA2 values in isolated tissues of SHR was alpha1/alpha2 > 5-HT2A > beta1. Similarly, we found that the in vitro functional activity of eugenosedin-A is quite different between WKY and SHR. On the other hand, in the isolated rabbit ear artery sensitized with 16 mmol/l K+, eugenosedin-A antagonized 5-nonyloxytryptamine- and serotonin-induced vasocontractions, indicating that it also blocked 5-HT1B and 5-HT2A receptors. In radioligand binding experiments, eugenosedin-A had significant binding affinities on alpha1/alpha2, beta1, 5-HT1B and 5-HT2A receptors. Finally, we suggest that the hypotensive effects of eugenosedin-A can be attributed to its multiple actions on the blockade of 5-HT1B, 5-HT2A, alpha and beta1 receptors in both WKY and SHR strains. PMID- 15118349 TI - Effects of phosphodiesterase V inhibition on nitric oxide-mediated relaxation responses in guinea pig trachea. AB - In the present study, we aimed to evaluate the effects of PDE V inhibition on NO mediated relaxation responses in isolated guinea pig trachea. Under the NANC conditions, tracheal preparations were contracted with histamine (100 microm/l). When contraction had reached a plateau, relaxation responses to electrical field stimulation (EFS, 60 V, 0.5 ms, 5-10 Hz) were determined before and after incubation of the tracheal ring with L-NAME (1 mmol/l), a NO synthase inhibitor. L-NAME significantly inhibited the relaxation responses and this inhibitory effect was reversed by L-arginine (1 mmol/l), a precursor of NO, but was not affected by D-arginine. In addition, cumulative application of the NO donors, 3 morpholino-sydnonimine (SIN-1) and sodium nitroprusside (SNP), caused concentration-dependent relaxation of tissues precontracted with histamine. The selective PDE type V inhibitor zaprinast at EC50 concentration (30 micromol/l) significantly potentiated EFS-induced NANC relaxations and relaxant responses to SIN-1 and SNP. In conclusion, these data support the hypothesis that NO is a mediator of NANC relaxations of guinea pig tracheal rings and PDE V inhibition potentiates NO-mediated relaxation. PMID- 15118350 TI - Mechanism of heme oxygenase-1 gene induction by quercetin in rat aortic smooth muscle cells. AB - We previously reported that adenovirus-mediated gene transfer of heme oxygenase-1 (HO-1) inhibits the development of atherosclerosis in apolipoprotein E-deficient mice. This finding implies that HO-1 induction is beneficial for protecting blood vessels. We also found that quercetin, a common polyphenolic compound in foods of plant origin, induces HO-1 expression in RAW264.7 cells. This study was aimed at examining the potency of quercetin as a HO-1 inducer and its regulation in rat aortic smooth muscle cells (RASMCs). We showed that quercetin-induced HO-1 production was in time- and dose-dependent fashions, and that this regulation occurred at both transcription and translation levels. Quercetin increased p38 mitogen-activated protein kinase (p38MAPK), but inhibited extracellular signal regulated kinase in RASMCs. The level of quercetin-induced HO-1 expression was attenuated by SB202190 (a p38MAPK inhibitor). Taken together from the data in this study, we suggest that quercetin induced HO-1 expression, at least in part, through p38MAPK. PMID- 15118352 TI - Association between norepinephrine transporter gene polymorphism and major depression. AB - Noradrenergic and serotonergic abnormalities have long been implicated in patients with major depression. The novel selective norepinephrine reuptake inhibitor reboxetine has been shown to be at least as effective as imipramine, desipramine and fluoxetine in the treatment of major depression. It is suggested that the dysfunction of the norepinephrine transporter (NET) may be related to major depression. Although the transcriptional activity related to the NET gene expression is little known, it may be a good candidate gene for major depression. Therefore, we investigated whether the T-182C polymorphism of the NET gene is associated with major depression in a Korean sample of 112 major depression patients compared with 136 healthy controls. We found a significantly lower frequency in TT genotype in patients with major depression than in normal controls when the genotypes of T-182C polymorphism were classified into two groups: TT group versus TC + CC group (p = 0.019). This result suggests that the T-182C polymorphism in the NET gene might be associated with major depression. PMID- 15118351 TI - Cytochrome P-450 2D6*10 C188T polymorphism is associated with antipsychotic induced persistent tardive dyskinesia in Chinese schizophrenic patients. AB - Typical antipsychotic treatment had been postulated to be a risk factor for the susceptibility to tardive dyskinesia (TD). The cytochrome P-450 debrisoquine/sparteine hydroxylase (CYP2D6) metabolizes a majority of antipsychotics and exhibits various phenotypes on enzymatic activities from poor metabolizers to ultrarapid metabolizers. The various phenotypes are encoded by polymorphic genetic variants on the CYP2D6 gene. Although several studies had explored the association between the CYP2D6*10 C188T polymorphism, which encodes the phenotype intermediate metabolizers, and TD in Orientals, the findings were inconclusive. In the present study, we examined the relationship between the CYP2D6*10 C188T polymorphism and the TD occurrence in 216 Chinese schizophrenic patients (113 patients with TD and 103 patients without TD) and explored the correlation between the TD severity assessed by the Abnormal Involuntary Movement Scale (AIMS) and each C188T genotype in the 113 TD patients. Using logistic regression analysis, we found a modest association (p = 0.045) between TD and C188T genotypes. This positive finding was only observed in male patients (p = 0.001), but not in females. Our findings also support the correlation between AIMS scores and C188T polymorphism within the TD group after adjusting for confounding effects with the multiple regression analysis (p = 0.033). We concluded that the CYP2D6*10 C188T polymorphism may be associated with the susceptibility to the occurrence of TD induced by typical antipsychotics, especially in male patients, and may also be correlated with AIMS scores in TD patients. PMID- 15118353 TI - Association study of a brain-derived neurotrophic factor (Val66Met) genetic polymorphism and panic disorder. AB - Brain-derived neurotrophic factor (BDNF), a member of the neurotrophic factor family, plays an important role in the development, maintenance and function of several neuronal systems. Recent studies have demonstrated that antidepressants, commonly used for panic disorder treatment, can increase central BDNF. In addition, animals with BDNF deficits have higher levels of anxiety when exposed to stressors in comparison to normal controls. The present study tested the hypothesis that the BDNF gene Val66Met polymorphism is associated with panic disorder. In this study, therefore, the incidence of this polymorphism was compared in 103 panic disorder patients and 180 normal controls. The genotype and allele frequencies for the BDNF gene Val66Met polymorphism did not differ comparing the two groups. Furthermore, no association was demonstrated between this BDNF polymorphism and either mitral valve prolapse or agoraphobia in panic disorder patients. These findings suggest that the investigated BDNF polymorphism does not play a major role in the pathogenesis of panic disorder in this Chinese population. Further studies exploring the relationship between genetic variations of BDNF and the cerebral atrophy associated with, and antidepressant treatment response in, panic disorder may be appropriate. PMID- 15118354 TI - No association of a functional polymorphism in the serotonin transporter gene promoter and anxiety-related personality traits. AB - Serotonergic neurotransmission, which is involved in many psychiatric disorders, is mediated by the serotonin transporter protein. Gene coding for the serotonin transporter protein is designated SLC6A4, which has been differentially associated with anxiety-related behavioral traits and neuroticism in healthy subjects. To confirm the association between the serotonin transporter (5-HTT) gene-linked polymorphic region (5-HTTLPR) and anxiety-related personality traits, we examined 228 healthy unrelated participants (age 38.6 +/- 12.8 years; 115 male, 113 female) of German origin, who were carefully examined with respect to psychiatric health. The self-ratable State-Trait Anxiety Inventory (STAI) and the NEO Five Factor Inventory (NEO-FFI) were performed. No significant association was observed between the 5-HTTLPR genotype and STAI 1 (state, chi2 = 0.82, p < 0.66, d.f. = 2), STAI 2 (trait, chi2 = 2.7, p < 0.25, d.f. = 2) and NEO-FFI scores of any of the 5 major axes, including neuroticism (chi2 = 3.35, p < 0.18, d.f. = 2) in all subjects. Given the small effect of this 5-HTT polymorphism on behaviour in previous studies, a lack of significant genotype differences in these tests could be due to considerable individual variability in these measures. PMID- 15118355 TI - BanI polymorphism of the cytosolic phospholipase A2 gene and mood disorders in the Korean population. AB - Membrane phospholipid abnormalities have been proposed to be involved in the pathogenesis of mood disorders, and in signal transduction and neurotransmitter uptake. Cytosolic phospholipase A2 (cPLA2) is not only an essential enzyme in the metabolism of fatty acids but also in signaling process. Therefore, we examined the association between the BanI polymorphism of the cPLA2 gene and mood disorders. Sixty-two patients with major depressive disorder (MDD), 50 patients with bipolar I disorder (BID) and 117 healthy controls participated in this study. Genotyping was performed by using PCR-based methods. Genotype and allele distributions in MDD patients were significantly different from those of the controls. In particular, the A2 allele was associated with increased risk of MDD development (p = 0.007, odds ratio = 1.827; confidence interval = 1.141-2.927). However, the polymorphism was not different between BID patients and controls in genotype and allele distribution. This preliminary study indicates the need for further studies on the potential role of the cPLA2 gene polymorphism in the susceptibility to mood disorders. PMID- 15118356 TI - Altered 5-HT2A and 5-HT4 postsynaptic receptors and their intracellular signalling systems IP3 and cAMP in brains from depressed violent suicide victims. AB - Serotonin 5-HT2A and 5-HT4 binding parameters and their second messengers 1,4,5 inositol triphosphate (IP3) and cyclic adenosyl monophosphate (cAMP) were studied in the frontal cortex, hippocampus, caudate nucleus and amygdala of 19 control subjects and 19 antidepressant-free, violent suicide victims. A significantly higher number of 5-HT4 receptors and higher second messenger cAMP concentrations were found in the frontal cortex and caudate nucleus of the depressed suicide victims as compared with the control group. Furthermore, significantly increased 5-HT2A binding sites and IP3 concentrations were noted in the caudate nucleus of the suicide victims, together with a significantly reduced number of 5-HT2A binding sites, higher binding affinity and increased IP3 concentrations in the hippocampus. No significant alterations in 5-HT4 and cAMP or in 5-HT2A and IP3 concentrations were observed in the amygdala. The caudate nucleus of depressed suicide victims seems to be the brain region with the highest alteration of the serotonergic system, and hence with the most diagnostic sensitivity. Further studies on suicidality and depression should focus on the functionality of the caudate nucleus. PMID- 15118357 TI - Association study of a functional catechol-O-methyltransferase genetic polymorphism with age of onset, cognitive function, symptomatology and prognosis in chronic schizophrenia. AB - The gene coding for catechol-O-methyltransferase (COMT), which is involved in the metabolism of catecholamines, has long been implicated as a candidate gene for schizophrenia. This study aimed to assess the relationship between a functional polymorphism (Val158Met) of the COMT gene and age of onset (AOO), symptomatology, global cognitive function and prognosis in patients with schizophrenia. The study enrolled 154 patients with schizophrenia from chronic wards. Results failed to show a significant association between the Val158Met polymorphism and the Brief Psychiatric Rating Scale scores, Mini-Mental State Examination scores, and Social and Occupational Functioning Assessment Scale scores, but COMT Val158Met heterozygotes had a later AOO than homozygous patients. However, by further expanding the number of patients to 228 patients, the differences in AOO among the three COMT genotypic groups was not significant. The COMT Val158Met polymorphism did not appear to significantly affect susceptibility, symptomatology, global cognitive function and prognosis in Chinese patients with schizophrenia, but the possible association with AOO merits further investigation. PMID- 15118358 TI - Quetiapine treatment for behavioral and psychological symptoms in patients with senile dementia of Alzheimer type. AB - The purpose of this study was to assess the effect of quetiapine in the treatment of behavioral and psychological symptoms of dementia (BPSD) in patients with senile dementia of Alzheimer type (SDAT). Sixteen SDAT patients with BPSD were recruited and quetiapine (25- 200 mg/day) was prescribed for 8 weeks. BPSD were evaluated with the Behavioral Pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD) and Cohen-Mansfield Agitation Inventory (CMAI) at week 0 (baseline) and week 8 (endpoint). The severity of the extrapyramidal symptoms was also assessed by the Drug-Induced Extrapyramidal Symptoms Scale (DIEPSS) at baseline and endpoint. Significant improvements were seen in the CMAI total score and in the BEHAVE-AD subscales of delusions, activity disturbances, aggressiveness, diurnal rhythm disturbances and in the BEHAVE-AD overall severity. There was no significant difference between the baseline and endpoint in the DIEPSS score. These data indicate that quetiapine is effective in controlling BPSD with favorable adverse-event profiles. PMID- 15118359 TI - Declarative memory impairments following a military combat course: parallel neuropsychological and biochemical investigations. AB - The aim of this study was to investigate the impact on several forms of memory and metabolism of a 5-day combat course including heavy and continuous physical activities and sleep deprivation. Mnemonic performance and biochemical parameters of 21 male soldiers were examined before and at the end of the course. Our results showed that short-term memory (memory span, visual memory, audiovisual association) and long-term memory were significantly impaired, whereas short-term spatial memory and planning tasks were spared. Parallel biochemical analysis showed an adaptation of energy metabolism. The observed decrease in glycaemia may be partly responsible for the long-term memory impairment, whereas the decreases in plasma cholinesterases and choline may be involved in the short-term memory deterioration. However, there are also many other reasons for the observed memory changes, one of them being chronic sleep deprivation. PMID- 15118360 TI - Nonconvulsive status epilepticus during cephalosporin therapy. AB - Cephalosporins may induce nonconvulsive status epilepticus (NCSE), a potentially reversible condition. Despite the wide use of these antibiotics, there are only few reported cases, because this condition is probably underestimated. We report two new cases of NCSE occurring during treatment with cefepime and ceftazidime, and emphasize the utility of emergent electroencephalogram in patients with an acute altered state of consciousness while receiving treatment with cephalosporins, particularly when there is evidence of impaired renal function. PMID- 15118361 TI - Protective effect of quercetin on the evolution of cisplatin-induced acute tubular necrosis. AB - BACKGROUND: The mechanism of cisplatin-induced nephrotoxicity is unknown, but has been associated with renal lipid peroxidation. The bioflavonoid quercetin may be a potential alternative to reduce cisplatin-induced nephrotoxicity. The aim of this study was to evaluate the effect of quercetin on the evolution of cisplatin induced acute tubular necrosis. METHODS: One hundred and three male Wistar rats were injected with cisplatin (5 mg/kg, i.p.), 43 of them received quercetin (50 mg/kg, by gavage) before cisplatin injection. Blood and urine were collected 5 and 20 days after the injection for the determination of plasma creatinine, urine volume and osmolality. The kidneys were removed for the determination of renal malondialdehyde (MDA) and for histological and immunohistochemical studies. The renal expression of fibronectin, alpha-smooth muscle actin, vimentin, Jun N terminal kinase, nuclear factor-kappaB, and macrophages during the evolution of the acute tubular necrosis induced by cisplatin and the histological changes observed in the kidneys were analyzed. RESULTS: Cisplatin-treated rats presented a transitory increase in plasma creatinine levels, tubular cell necrosis and increased immunostaining for vimentin, alpha-SM-actin, fibronectin, ED1, NF kappaB, and p-JNK in the renal cortex and outer medulla. These alterations were less intense in animals treated with quercetin. CONCLUSION: Quercetin treatment attenuated the functional, histological and immunohistochemical alterations induced by cisplatin. PMID- 15118365 TI - Albert Montgomery Kligman. MD, PhD, Dr.h.c.mult., Professor of Dermatology. PMID- 15118362 TI - Protein methylation activates reconstituted ryanodine receptor-ca release channels from coronary artery myocytes. AB - Ryanodine receptors (RyR) play an important role in the regulation of intracellular Ca(2+) concentration and in the control of vascular tone. However, the mechanism regulating the activity of RyR is poorly understood. The present study determined whether protein methylation participates in the control of RyR activity. Using a planar lipid bilayer clamping system, S-adenosyl-L-methionine (SAM), a methyl donor, significantly increased the activity of a 245-pS reconstituted Ca(2+) release channel from coronary arterial smooth muscle (CASM) in a concentration-dependent manner. Addition of the protein methylation blockers, 3-deazaadenosine, S-adenosylhomocysteine or sinefungin into the cis solution markedly attenuated SAM-induced activation of RyR/Ca(2+) release channels. By Western blot analysis, arginine N-methyltransferase (PRMT1) and FK506 binding protein (FKBP) were detected in the SR used for reconstitution of RyR. In the presence of anti-PRMT1 antibody (1:100), SAM-induced activation of RyR/Ca(2+) channel was completely abolished. In addition, this SAM-induced increase in RyR/Ca(2+) channel activity was blocked by 30 microM ryanodine and by FK506 (100 microM), a ligand for the RyR accessory protein. These results suggest that protein methylation activates RyR/Ca(2+) release channels and may participate in the control of intracellular Ca(2+) mobilization in CASM cells by transferring a methyl group to the arginine moiety of the RyR accessory protein, FKBP 12. PMID- 15118363 TI - Arteriolar endothelial dysfunction is restored in ischaemic muscles by chronic electrical stimulation. AB - Chronic intermittent electrical stimulation (15 min on, 85 min off, seven times per day) eliminated endothelial dysfunction of pre-capillary arterioles in ischaemic rat ankle flexor muscles. Responses to acetylcholine were restored from constriction to dilation, and the reduced dilation to bradykinin was corrected by 1 week of stimulation. Administration of the NOS inhibitor N(omega)-nitro-L arginine for 1 week impaired arteriolar reactivity in a similar way to ischaemia, and dilator function was likewise restored by chronic stimulation. This suggests that nitric oxide production in the microcirculation is depressed by chronic ischaemia and that chronic electrical stimulation can specifically reverse this deficit. Stimulation applied to ischaemic muscles for 2 weeks also increased the numbers of microvessels immunostained for alpha-smooth muscle actin and the numbers of eNOS-positive microvessels and capillaries. These findings help to elucidate the mechanism of the beneficial effect of exercise in the treatment of peripheral vascular diseases by showing that muscle activity can improve both function and structural capacity of the microvasculature. PMID- 15118366 TI - A personal critique on the state of knowledge of Rosacea. PMID- 15118367 TI - Pentoxifylline in the treatment of actinic prurigo. A preliminary report of 10 patients. AB - BACKGROUND: Actinic prurigo (AP) is a chronic familial photodermatosis usually seen in Latin-American Mestizo and Indian populations. It frequently begins in childhood and is more prevalent in females. The pathogenesis of AP has not been clearly elucidated, but previous studies have suggested an immune-mediated condition. Many drugs have been employed to treat AP patients with variable success. OBJECTIVE: To assess the clinical efficacy of pentoxifylline (PTX) in the treatment of AP patients by measuring its effects on lesions and pruritus. METHODS: 10 patients with severe AP were included to receive PTX in a 6-month open-label uncontrolled study. RESULTS: Clinical improvement of lesions was evident in all patients. Relief in pruritus was evident after 1 month of treatment and was maintained while receiving PTX. Five patients were followed up for 2 years, 2 obtained complete remission, and 3 had an important reduction in the use of corticosteroids. CONCLUSION: PTX was useful in the treatment of our actinic prurigo patients. It may induce a complete or partial remission of lesions and allow a decrease in the use of topical corticosteroids. PMID- 15118368 TI - Diagnostic and surgical accuracy and economic aspects of dermatological surgery - a pilot study. AB - BACKGROUND: Good skin care for oncological dermatological patients includes efficacious excision to achieve tumour control and economically reasonable costs. This field concerns dermatologists as much as other surgical specialities. METHODS: Of 944 excisions and biopsies, we studied 114 epidermal carcinomas excised by plastic surgeons and dermatologists. This allowed us to compare the accuracy concerning oncological surgical aspects as well as the extrapolated costs produced by these two specialities. RESULTS: Dermatologists are significantly more accurate concerning total excision of epidermal tumours compared to plastic surgeons. CONCLUSION: From an economic point of view, plastic surgeons are increasingly more expensive than dermatologists. Most expenses are due to the use of a hospital operating room. PMID- 15118369 TI - Systematic overview of the pharmacological management of postherpetic neuralgia. An evaluation of the clinical value of critically selected drug treatments based on efficacy and safety outcomes from randomized controlled studies. AB - OBJECTIVE: This study systematically reviews current evidence on drug treatments commonly used in postherpetic neuralgia. METHODS: Randomized controlled trials were critically selected using predefined search criteria. Efficacy was evaluated as percentage of improvement in pain intensity between baseline and endpoint, tolerability by number of study discontinuations because of adverse events and incidence of adverse events. RESULTS: Currently published trials enrolled different patient populations and small patient numbers. The great variability in doses, titration schemes, designs and washout periods together with other design flaws made comparison between different studies scientifically impossible. CONCLUSIONS: There is a real need for well-performed clinical trials with standardization in design and reporting. Development of adequate and validated questionnaires for evaluation and comparison of efficacy and safety of treatments is also needed. Based on the evaluation of individual studies, it is concluded that only gabapentin is studied in large (over 200 patients), placebo-controlled studies showing good efficacy and safety. PMID- 15118370 TI - c-kit Mutation in generalized lentigines associated with gastrointestinal stromal tumor. AB - We describe the case of a 59-year-old Japanese woman presenting with generalized lentigines without systemic anomalies. She had a medical history of gastrointestinal stromal tumors (GISTs), in which gain-of-function mutations of the c-kit gene had recently been found. We detected a point mutation at codon 557 in exon 11 of leukocyte DNA from the patient. The stem cell factor-type III receptor tyrosine kinase pathway plays important roles in the regulation of melanocyte proliferation and differentiation. We speculate that the generalized lentigines of the patient may be caused by melanocyte proliferation due to the c kit gene mutation. PMID- 15118371 TI - CCR4 Expression by atypical T cells in systemic pilotropic lymphoma: its behavior under treatment with interferon gamma, topical PUVA and systemic retinoid. AB - We describe an 88-year-old Japanese patient with pilotropic T cell lymphoma involving the peripheral blood as well as lymph nodes. This patient presented with multiple red follicular papules, confluent, infiltrated erythematous plaques and nodules. Moreover, he was conspicuous for the presence of total alopecia of the scalp and eyebrows. Histopathologically, the lesional skin showed dense follicular and perifollicular infiltrates of atypical lymphocytes. The flow cytometry disclosed the presence of weakly CD4+ CCR4+ cell populations that would not be detected in the peripheral blood from healthy controls. The patient responded well to topical PUVA and systemic etretinate (retinoid-PUVA) and intravenous IFN-gamma. Parallel with the decrease in atypical cells in the peripheral blood, the percentage of weakly CD4+ CCR4+ T cells declined. However, about 1 week after we discontinued this treatment because of the side effects, the lymph node swelling became prominent, and, 4 weeks later, the patient died before restarting any specific chemotherapy. PMID- 15118372 TI - Folliculitis decalvans associated with Micronychia. PMID- 15118373 TI - Eosinophilic pustular folliculitis with a butterfly rash-like distribution. PMID- 15118374 TI - Cutaneous angiomyolipoma. PMID- 15118375 TI - Photodistributed lichenoid drug eruption with rhabdomyolysis occurring during leflunomide therapy. PMID- 15118376 TI - Cafe-au-lait macules: are these markers of neoplasia of genetic origin? PMID- 15118377 TI - Fixed drug eruption of the penis due to zolmitriptan. PMID- 15118378 TI - Use of duplex ultrasonography in the treatment of thromboangiitis obliterans with iloprost. AB - We present a 34-year-old patient with digital necrosis due to thromboangiitis obliterans. He was successfully treated with iloprost, a prostaglandin analogue. Duplex ultrasonography was performed during the perfusion of iloprost to optimize the doses and the treatment duration. A complete revascularization was observed after 10 days. Iloprost perfusions were stopped, and a slow regression of the necroses was observed in the subsequent days. With the use of duplex ultrasonography, unnecessary high doses of iloprost and long periods of treatment can be avoided reducing side effects and treatment costs. PMID- 15118379 TI - Dermoscopy for the in vivo detection of sarcoptes scabiei. AB - We report the case of an 80-year-old patient who had intense pruritus which did not respond to a 3-month treatment with topical corticosteroids. On dermoscopy examination of the excoriations, we found the typical dermoscopic aspect of the scabies mite at a distance. Dermoscopy allows identifying a triangular structure which corresponds to the anterior section of the mite including the mouth part and the 2 pairs of front legs. This aspect has been described as resembling a jetliner with its trail, a delta glider or a spermatozoid. Traditional diagnostic methods for scabies failed in this case because the mites were at a distance from the burrows. This was due to the fact that the reaction to the mite was less pronounced and the diagnosis is frequently missed. Dermoscopy is a useful tool for the diagnosis of scabies either as a diagnostic test or to guide the traditional diagnostic tests. PMID- 15118381 TI - Progressive respiratory failure in paraneoplastic pemphigus associated with chronic lymphocytic leukemia. AB - A 70-year-old Caucasian man with chronic lymphocytic leukemia suffered from widespread, histologically proven cutaneous lichen planus responding to topical corticosteroids. 2 years later, he presented with painful erosive stomatitis and increasing dyspnea. Histology, direct and indirect immunofluorescence were diagnostic for paraneoplastic pemphigus. A full diagnostic workup could not disclose the cause of the progressive respiratory insufficiency. Despite aggressive treatment of the lymphocytic leukemia and the paraneoplastic pemphigus, the patient died 3 months after diagnosis. Paraneoplastic pemphigus may lead to pulmonary failure which is refractory to treatment and has a fatal outcome. PMID- 15118380 TI - First report of Arthroderma benhamiae in Switzerland. AB - BACKGROUND: Dermatophytes are usually identified on the basis of macroscopic characteristics and microscopic examination of the cultures. Identification of dermatophytes often remains difficult or uncertain because there are variations from one isolate to another and overlapping characteristics between species. OBJECTIVE: To identify dermatophyte species producing numerous microconidia and resembling Trichophyton mentagrophytes by DNA sequence analysis. METHODS: The complete ITS1 + 5.6s + ITS2 rDNA region of various dermatophytes isolated in culture was amplified by PCR and sequenced. RESULTS: Nine isolates of a fast growing dermatophyte species were identified as Arthroderma benhamiae by DNA sequencing. Retrospective investigations revealed that the isolates were from 8 children and 1 adult suffering from inflammatory dermatophytosis. Eight of the 9 patients had had previous contact with rodents, mostly guinea pigs. CONCLUSION: It is the first time that A. benhamiae is reported in Switzerland. In cases of dermatophytosis attributed to A. benhamiae, a rodent is the most likely cause of infection. PMID- 15118382 TI - Micronodular Kaposi's sarcoma - a new variant of classic-sporadic Kaposi's sarcoma. AB - Kaposi's sarcoma (KS) in its classic-sporadic form is a rare vascular neoplasm affecting predominantly elderly men of Jewish or Mediterranean origin. Women are very rarely affected by KS. The tumor manifests itself most commonly with brownish macular or infiltrated oval lesions on the lower legs in both genders. Human herpesvirus 8 (HHV-8) has recently been demonstrated to be etiologically linked to KS. We report a 51-year-old HIV-seronegative Caucasian woman with an unusual, previously not described variant of KS which presented with small (3 mm in diameter) scattered firm red papules on her right arm. The histology with nodular spindle cell tumor expressing endothelial markers, the demonstration of serum antibodies against HHV-8 and the presence of viral nucleic acids in the lesional tissue proved the diagnosis of the nodular stage of KS. An indolent course characterized by clinically identical seasonal recurrences, even after surgical treatment and cryosurgery, was observed during the follow-up period of 9 years. We propose the term 'micronodular KS' for this unusual clinical variant of KS mimicking capillary hemangioma. PMID- 15118383 TI - Subcutaneous infection with Mycobacterium abscessus in a renal transplant recipient. AB - Transplant recipients show a high incidence of infections, often with atypical manifestations mainly because of drug-related immunosuppression. Mycobacterial infections can be life-threatening in immunosuppressed patients due to the risk of dissemination. There have been only few reports of cutaneous infections with Mycobacterium abscessus among immunosuppressed patients. We present a rare case with a sporotrichoid skin infection with M. abscessus in a renal recipient. The value of immunohistochemical detection of mycobacteria using an anti-BCG antibody in the diagnostic work-up skin biopsies is demonstrated. Long-term antibiotic treatment resulted in complete remission, although recurrences with atypical presentation were observed. PMID- 15118385 TI - Pancreatic panniculitis in a patient with an acinar cell cystadenocarcinoma of the pancreas. AB - We describe a 60-year-old man with pancreatic panniculitis associated with arthritis and peripheral eosinophilia in whom the skin symptoms led to a diagnosis of an underlying acinar cell cystadenocarcinoma. The panniculitis involved initially the legs, but soon thereafter lesions developed on the trunk and upper extremities. In the literature, only 5 cases of pancreatic acinar cell cystadenocarcinoma have been reported, none of these in association with panniculitis. PMID- 15118384 TI - Purpura fulminans in a child as a complication of chickenpox infection. AB - Purpura fulminans is a thrombotic disease that can occur during infections, disseminated intravascular coagulation or in the context of an acquired or congenital protein C or S deficiency. Here we report the case of a 4-year-old child who developed, 5 days after a chickenpox infection, large painful ecchymotic, necrotizing and retiform plaques on the lower extremities. Laboratory analyses revealed very low protein S levels as well as anticardiolipin antibodies. Aggressive treatment by low-molecular-weight heparin, steroids, intravenous immunoglobulins and fresh frozen plasma was able to prevent the extension of the lesions and to correct the coagulation abnormalities. No lesions required skin grafting. As in our patient, an acquired protein S deficiency is probably responsible for most cases of purpura fulminans occurring after varicella, but the concomitant presence of antiphospholipid antibodies may also play a role. PMID- 15118386 TI - Cutaneous myiasis due to Dermatobia hominis. AB - Cutaneous myiasis caused by the human botfly Dermatobia hominis involves the infestation of tissue with dipterous fly larvae and is common in the neotropical region of the New World. We report a case of D. hominis imported in Switzerland from Costa Rica. In the past, various approaches to extract the botfly larva have been reported. PMID- 15118387 TI - Giant fibrokeratoma of the heel. AB - We describe a 77-year-old patient with a giant acquired fibrokeratoma on the heel. The size and the localization of the tumor was unusual. Simple shave excision was curative. PMID- 15118388 TI - Localized childhood vulval pemphigoid treated with tacrolimus ointment. AB - BACKGROUND: Localized vulval childhood pemphigoid is a rare variant within the pemphigoid group. Although its prognosis seems favorable, the best therapeutic strategy remains unclear. OBSERVATION: We here describe the case of an 8-year-old girl presenting with a 5-year history of relapsing vulval pain and lesions suggestive of lichen sclerosus. Clinical features, light microscopy and direct immunofluorescence microscopy were consistent with vulval cicatricial pemphigoid, although the autoantigen(s) involved could not be characterized. Her disease responded to treatment with topical tacrolimus ointment 0.1% within 3 months without any evidence for disease activity, except for slight residual scarring. After 12 months, her treatment was stopped without relapse. CONCLUSION: This observation suggests that in this rare immune-mediated blistering disease topical tacrolimus is an interesting therapeutic option without the adverse effects associated with topical steroids. PMID- 15118389 TI - Lupus-like syndrome associated with statin therapy. AB - Statins are among the most widely prescribed drugs. An increasing number of lupus like syndrome has recently been reported with these lipid-lowering agents. We describe a new case associated with simvastatin therapy. The presence of anti dsDNA antibodies in the serum is for the first time reported confirming that statins may also induce a systemic autoimmune reaction. Statin-induced lupus-like syndrome is characterized by the long delay between the beginning of therapy and the skin eruption. Antinuclear antibodies may persist for many months after drug discontinuation. The causal relationship may be therefore difficult to establish, and probably many cases are unrecognized. Early diagnosis may avoid unnecessary immunosuppressive therapy. PMID- 15118390 TI - Nephrogenic fibrosing dermopathy treated with extracorporeal photopheresis. AB - Nephrogenic fibrosing dermopathy (NFD) is a rare fibrosing skin disease of unknown etiology occurring in patients with terminal renal disease. It was first described in the year 2000. The histology of NFD shows an increased number of dendritic cells, fibroblasts and thickened collagen fibers resembling scleromyxedema. It can be distinguished from scleromyxedema by a different distribution pattern of the skin lesions with indurated plaques mainly on the extremities and the absence of paraproteinemia. As yet, no treatment for NFD has been proven to be uniformly efficient. We describe the case of a 40-year old patient with renal insufficiency who was treated with hemodialysis and who had undergone kidney transplantation. Two years after transplantation, she developed sclerodermiform brownish plaques on her extremities. The induration improved significantly after 4 cycles of extracorporeal photopheresis. PMID- 15118391 TI - Familial cutaneous mycosis fungoides: successful treatment with a combination of gemcitabine and alemtuzumab. AB - We report a familial cutaneous T-cell lymphoma in father and son. After different treatment modalities without lasting responses, the son was treated with gemcitabine as single agent and due to insufficient effect with alemtuzumab monotherapy. Only after the two drugs had been combined did we observe a remarkable response of the skin lesions and disappearance of enlarged lymph nodes. The combined treatment with gemcitabine and alemtuzumab was well tolerated, and no increased toxicity was noted. The combination of these two active agents may provide an additional option in the treatment of cutaneous T cell lymphoma. PMID- 15118392 TI - Fulminant herpetic sycosis: atypical presentation of primary herpetic infection. AB - Fulminant herpetic sycosis is a rare but well-known manifestation of herpes simplex virus (HSV) infection occurring in the context of viral recurrence in immunodepressed patients. We present here the case of a 32-year-old male patient, without notable medical history, who developed papulovesicular lesions of the beard accompanied by fever, painful cervical lymphadenopathy and odynophagia, with a clinical evolution that was initially unfavourable under antibiotic treatment. The diagnosis of herpetic sycosis was established by means of direct immunofluorescence and culture which confirmed positivity for HSV-1 and serologies compatible with a primary viral infection. No sign for a latent immune deficit was found at the time of investigations. The clinical evolution was rapidly favourable with administration of intravenous aciclovir for 1 week. To our knowledge, herpetic sycosis as a presentation of primary viral infection has not been reported previously. The possibility of a herpetic sycosis of the beard must be considered in the case of non-response to antibiotic or antifungal treatment. PMID- 15118393 TI - Cutaneous plasmablastic lymphoma in an HIV-positive male: an unrecognized cutaneous manifestation. AB - Plasmablastic lymphoma (PBL) is a rare and relatively new entity originally described in HIV-infected individuals. This subset of Epstein-Barr-virus (EBV) related non-Hodgkin lymphomas is now regarded as a distinct clinicopathological category of AIDS-associated lymphomas occurring preferentially in the oral cavity and showing a poor prognosis. We describe for the first time an EBV-associated PBL with an isolated cutaneous distribution on the lower extremities in an HIV infected heterosexual male and point to the unique clinical, morphological and immunophenotypic characteristics of this lymphoma. The patient presented with fast growing solid and livid nodules on both legs. The large, blastic tumor cells showed the following immunophenotype: CD138+, CD45+, CD20-, CD10-, CD3-, CD30-, bcl-2-, bcl-6-, LMP-1- and EMA-. The proliferation fraction (Mib-1) was >90%. EBV association was demonstrated by in situ hybridization (EBV-encoded RNAs 1/2). Polymerase-chain-reaction-based DNA analysis demonstrated a clonal IgH rearrangement in the absence of a bcl-2/IgH translocation. PBL in HIV patients may occur not only in the oral cavity, but can probably involve any other organs including the skin. PMID- 15118394 TI - Technologies in the evaluation of irritable bowel syndrome. AB - During a meeting in The Hague, The Netherlands, the IBiS Club evaluated the most important techniques that can be used in the investigation of irritable bowel syndrome, either in the context of scientific research or as a clinical diagnostic tool. In each of these, the relevance of findings made in irritable bowel syndrome was balanced against the applicability of the technique. The discussion of the group is summarized in this paper. PMID- 15118395 TI - Extended microsatellite analysis in microsatellite stable, MSH2 and MLH1 mutation negative HNPCC patients: genetic reclassification and correlation with clinical features. AB - BACKGROUND: Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant disorder predisposing to predominantly colorectal cancer (CRC) and endometrial cancer frequently due to germline mutations in DNA mismatch repair (MMR) genes, mainly MLH1, MSH2 and also MSH6 in families seen to demonstrate an excess of endometrial cancer. As a consequence, tumors in HNPCC reveal alterations in the length of simple repetitive genomic sequences like poly-A, poly-T, CA or GT repeats (microsatellites) in at least 90% of the cases. AIM OF THE STUDY: The study cohort consisted of 25 HNPCC index patients (19 Amsterdam positive, 6 Bethesda positive) who revealed a microsatellite stable (MSS)--or low instable (MSI-L)--tumor phenotype with negative mutation analysis for the MMR genes MLH1 and MSH2. An extended marker panel (BAT40, D10S197, D13S153, D18S58, MYCL1) was analyzed for the tumors of these patients with regard to three aspects. First, to reconfirm the MSI-L phenotype found by the standard panel; second, to find minor MSIs which might point towards an MSH6 mutation, and third, to reconfirm the MSS status of hereditary tumors. The reconfirmation of the MSS status of tumors not caused by mutations in the MMR genes should allow one to define another entity of hereditary CRC. Their clinical features were compared with those of 150 patients with sporadic CRCs. RESULTS: In this way, 17 MSS and 8 MSI-L tumors were reclassified as 5 MSS, 18 MSI-L and even 2 MSI-H (high instability) tumors, the last being seen to demonstrate at least 4 instable markers out of 10. Among all family members, 87 malignancies were documented. The mean age of onset for CRCs was the lowest in the MSI-H-phenotyped patients with 40.5 +/- 4.9 years (vs. 47.0 +/- 14.6 and 49.8 +/- 11.9 years in MSI-L- and MSS phenotyped patients, respectively). The percentage of CRC was the highest in families with MSS-phenotyped tumors (88%), followed by MSI-L-phenotyped (78%) and then by MSI-H-phenotyped (67%) tumors. MSS tumors were preferentially localized in the distal colon supposing a similar biologic behavior like sporadic CRC. MSH6 mutation analysis for the MSI-L and MSI-H patients revealed one truncating mutation for a patient initially with an MSS tumor, which was reclassified as MSI L by analyzing the extended marker panel. CONCLUSION: Extended microsatellite analysis serves to evaluate the sensitivity of the reference panel for HNPCC detection and permits phenotype confirmation or upgrading. Additionally, it confirms the MSS status of hereditary CRCs not caused by the common mutations in the MMR genes and provides hints to another entity of hereditary CRC. PMID- 15118396 TI - Keratin 2e: a marker for murine nipple epidermis. AB - Mesenchyme-derived signals influence the unique keratinization and appendage formation programs in specialized skin regions. Interactions between primary mammary mesenchyme and epidermal cells result in the formation of the nipple; however, it is unclear whether this represents a site of regionally specialized epidermis. We profiled the ultrastructure and keratin expression of the murine nipple, and the ventral skin of the K14-parathyroid hormone-related protein (PTHrP) transgenic mouse, which models nipple formation. We found the murine nipple and ventral K14-PTHrP epidermis display expanded suprabasal and granular layers, as well as a thickened cornified layer compared to ventral skin of wild type littermates. We also observed increased levels of filaggrin in extracts from the ventral epidermis of the K14-PTHrP mouse when compared to that of wild-type littermates. Keratin 2e, previously reported to be expressed in various specialized epidermal sites in the mouse, is expressed in the nipple and the ventral skin of the K14-PTHrP mouse. Keratinocytes grown from the ventral epidermis of the K14-PTHrP mouse or wild-type littermates exhibited identical expression of epidermal markers in vitro, suggesting that the modulated differentiation profile observed in the nipple or the ventral K14-PTHrP skin was dependent on interactions with fibroblasts. The lack of appendages, altered stratification pattern and expression of a specialized keratin suggests that the murine nipple is an example of regionally specialized epidermis. PMID- 15118397 TI - Pattern of myosin heavy chain isoforms in different fibre types of canine trunk and limb skeletal muscles. AB - The aim of this study was to determine the pattern of myosin heavy chain (MHC) isoform expressions within the muscle fibres of functionally diverse trunk and limb dog muscles using monoclonal antibodies that are specific to MHC isoforms. We found that three MHC isoforms are expressed in dog skeletal muscles. The pattern of their expressions determined the existence of 'pure' fibres, i.e. I and IIa, both expressing only one MHC isoform, and 'hybrid' fibres, i.e. I/IIa and IIa/x, that co-expressed two MHC isoforms. While the MHCI, MHCIIa and MHCI/IIa fibres corresponded to the myofibrillar ATPase type fibres I, IIA and IIC, respectively, the hybrid MHCIIa/x fibres mostly behaved like the IIDog fibre type in myofibrillar ATPase reaction as described by Latorre et al. No pure MHCIIx fibres were found. Though MHCIIa/x fibres were quite numerous, their presence varied not only within different muscles but within the same muscle of different animals as well. We suggest that the discrepancies in the classification of fibre types according to their myofibrillar ATPase activity between different studies of dog skeletal muscles are probably a consequence of the variable content of the MHCIIa and MHCIIx isoforms in the MHCIIa/x hybrid fibres. Estimating the histochemical metabolic profile of fibres we found that in all fast fibres oxidative-glycolytic metabolism prevailed, whereas in slow fibres oxidative metabolism was more pronounced. PMID- 15118398 TI - Apocrine glands in the eyelid of primates contribute to the ocular host defense. AB - Apocrine glands of Moll are regular components of primate eyelids. We studied the distribution and localization of these glands in three different primate species, the common marmoset, the rhesus monkey, and the hamadryas baboon. In addition, we tested the primate glands of Moll with antibodies against antimicrobial proteins, cytoskeletal proteins and the androgen receptor. The glands of Moll differ in abundance and distribution in different monkeys. In the common marmoset, a representative of the New World monkeys, Platyrrhini, the apocrine glands are frequently found at the lid margin and in the overlying epidermis of the lid. In the rhesus monkey and the hamadryas baboon, representatives of Old World monkeys, Catarrhini, apocrine glands are rarer and located predominantly at the margin of the lid. The immunohistochemical analysis indicates the presence of a variety of antimicrobial proteins, e.g. lysozyme, beta-defensin-2, adrenomedullin, lactoferrin, and IgA, in these glands. Interestingly, there are basically no androgen receptors in the nuclei of apocrine glands at the lid margin in all three monkey species. In the common marmoset, however, androgen receptors are found in apocrine glands of the overlying epidermis of the lid. We speculate that the glands of Moll are derived from apocrine glands as found in the skin of the entire body in New World monkeys which developed at the lid margins of higher primates and humans into specialized glands secreting agents of host defense in the eye. PMID- 15118399 TI - Reevaluation of the morphological parameters according to 11 different duplications of the fetal vertebral artery at prevertebral (V1) and intracranial (V4) parts. AB - It is noted that the vertebral artery lies buried in a sea of generalities, prejudices, fear, and ignorance in the clinical literature. This fact is the reason why this observation is based on 11 cases, selected from 206 fetal cases, of various duplications of the vertebral artery. Aberrant or anastomotic vessels are illustrated and their definition is largely modified according to the recent anatomic terminology. Duplications of the vertebral artery indicate the 'segmental duplication' as a first form, and the presence of an abnormal 'fenestra' in the vessel's trunk as a second form of duplication. There are 9 (5 at prevertebral and 4 at intracranial parts) or 4.36% cases of 'segmental duplication' of the vertebral artery and 2 or 0.98% 'arteria vertebralis unifenestrata' at the intracranial part of all 206 cases. Successive morphological parameters of aberrant or anastomotic vessels are described and discussed according to personal cases and cases in the literature. PMID- 15118400 TI - Macro- and microanatomical characterization of the cat brachial plexus. AB - The anatomy of the cat brachial plexus is poorly documented despite its importance for reparative surgery. The present work aims to study the anatomy of the brachial plexus of 20 European cats and to analyze the radicular constituents of 10 of them using histological techniques. The plexus radicular constitution is very homogeneous and is mainly composed of the ventral rami (also called ventral branches) arising from C6, C7, C8 and T1 spinal nerves. The fascicular and axonal structure is more variable. C8 is the ventral ramus that contains the largest number of axons (16,673 +/- 3,307), and is composed of 6.7 +/- 3.5 fascicles. The radial nerve is the peripheral nerve that contains the largest number of axons (11,245 +/- 2,217), and is composed of 7.3 +/- 3.8 fascicles. The number of nervous fibers does not differ proportionally to the weight of the cat, or between the right or the left side. Our data could help neurotization and entubulation repair after brachial plexus injury. PMID- 15118407 TI - Efficiency, fidelity and enzymatic switching during translesion DNA synthesis. AB - More than half of the 16 human DNA polymerases may have some role in DNA replication and potentially modulate the biological effects of DNA template lesions that impede replication fork progression. As one approach to understand how multiple polymerases are coordinated at the fork, we recently quantified the efficiency and fidelity with which one particular translesion synthesis enzyme, human DNA polymerase eta, copies templates containing cis-syn thymine dimers. Several observations from that study were unanticipated. Here we discuss the structural and biological implications of those results in light of earlier studies of translesion synthesis. PMID- 15118408 TI - p53 designer genes for the modern mouse. AB - Major efforts are underway to develop molecular strategies that target the p53 pathway for the treatment of cancer. Mouse strains with humanized p53 sequences that present the precise human DNA-binding domain as mutation target could be informative models to test p53 rescue drugs, and to explore experimentally the causes of human tumor mutations. PMID- 15118409 TI - Maintenance of G1 checkpoint controls in telomerase-immortalized endothelial cells. AB - Here we report the characterization of a series of telomerase-immortalized human umbilical vein endothelial cell lines (i-HUVEC). These cells maintain endothelial characteristics such as marker expression, dependence on basic fibroblast growth factor for proliferation, and the ability to form tube structures on Matrigel. In addition, these cells do not show signs of tumorigenic transformation because their growth is contact-inhibited, serum-dependent, and anchorage-dependent. In addition, i-HUVEC do not grow or survive when implanted subcutaneously in immunocompromised mice. Notably, the i-HUVEC lines maintain normal p53-dependent checkpoint control, inducing expression of p21(Cip1/Waf1) in response to DNA damage. These cells subsequently decrease phosphorylation of pRb and arrest in G1. Furthermore, the i-HUVEC lines maintain normal p53-independent checkpoint control, inducing expression of p27(Kip1) in response to lovastatin treatment, with a subsequent decrease in pRb phosphorylation. Lovastatin-treated i-HUVEC lines undergo a G1 arrest that can be reversed with comparable kinetics to that of low passage HUVEC. Together these data demonstrate that telomerase immortalized endothelial cells can retain normal phenotypes and cell cycle regulation. This result could have significant implications in the study of angiogenic processes such as tumor growth, wound healing, and the vascularization of engineered tissue. PMID- 15118410 TI - The use of CDK inhibitors in oncology: a pharmaceutical perspective. AB - To date dozens of pharmacophores with CDK-inhibitory properties have been discovered and the first compounds possessing such activity are now under clinical evaluation. Because the real therapeutic utility of these agents should be able to be assessed in the foreseeable future, it is unfortunate that CDK biology remains a moving target. Until recently is has been held that inhibiting CDK2, presumed master of the 10 known CDK isoforms, should be most beneficial in terms of achieving potent and selective antiproliferative effects in transformed cells. Recent findings, however, suggest that CDK2 may not be the key cell cycle player previously assumed, after all. It has also now become clear that CDKs have functions in physiological processes other than coordination of cell cycle progression, particularly regulation of DNA transcription. The implications of these new biological insights for the discovery and development of CDK inhibitors in oncology are discussed. PMID- 15118411 TI - HAUSP is required for p53 destabilization. AB - p53 ubiquitination is the principal mechanism by which p53 levels are regulated in the cell. HAUSP (also known as USP7) has been proposed to serve as a substrate specific deubiquitinase of p53, and an increase in p53 levels was reported upon overexpression of HAUSP. We have disrupted the HAUSP genomic locus by homologous recombination and shown that HAUSP ablation results in a phenotype opposite to that predicted. Rather than decreasing p53 levels associated with increased p53 ubiquitination, the absence of HAUSP resulted in p53 accumulation accompanied by decreased p53 ubiquitination. The p53 protein in HAUSP-deficient cells was active, as assessed by the induction of its transcriptional targets and growth arrest. The basis for this phenotype was traced to the increased ubiquitination of MDM2, a negative regulator of p53 levels. These results demonstrate that MDM2, rather than p53, is the substrate for HAUSP under physiologic conditions and document a fascinating and unexpected twist to the regulation of the p53/MDM2 axis. PMID- 15118412 TI - Methods to classify BRCA1 variants of uncertain clinical significance: the more the merrier. PMID- 15118413 TI - An effective method for selecting siRNA target sequences in mammalian cells. AB - RNA interference is a gene-silencing phenomenon triggered by dsRNA (double stranded RNA) and has been widely used for studying gene functions. The short interfering RNA (siRNA) responsible for RNA interference, however, varies markedly in its gene-silencing efficacy. Because this efficacy depends on the selected target sequences, we developed an effective selection method based on the gene degradation measure (priority score) defined by positional features of individual nucleotides. We tested this method experimentally by using it to select new siRNA target sequences in the homo sapiens cyclin B1 gene (CCNB1) and confirmed that it selected highly effective gene-silencing sequences. The proposed method will therefore be useful for selecting new siRNA target sequences in mammalian cells. PMID- 15118414 TI - Multiple roles of canonical Wnt signaling in cell cycle progression and cell lineage specification in neural development. AB - Signaling by the canonical Wnt pathway has multiple functions in stem cells. It can either control stem cell expansion or--as we have recently demonstrated with neural crest stem cells-influence cell lineage decisions by promoting specific fates at the expense of others. Thus, the role of canonical Wnt in stem cells is dependent on cell--intrinsic properties that determine how a cell responds to Wnt. The molecular basis for the functional diversity of Wnt in different stem cell types remains to be elucidated. PMID- 15118415 TI - DCX's phosphorylation by not just another kinase (JNK). AB - The mammalian cortex is generally subdivided into six organized layers, which are formed during development in an organized fashion. This organized cortical layering is disrupted in case of mutations in the doublecortin (DCX) gene. DCX is a Microtubule Associated Protein (MAP). However, besides stabilization of microtubules, it may be involved in additional functions. The participation of this molecule in signal transduction is beginning to emerge via discovery of interacting molecules and its regulation by phosphorylation using several different kinases. We raise the hypothesis, that the combinatorial phosphorylation of DCX by different kinases and at different sites may be a molecular regulatory switch in the transition of a migrating neuron through multiple phases of migration. Our recent research has suggested the involvement of DCX in the JNK (Jun-N-terminal Kinase) pathway. The JNK pathway is linked to the reelin pathway, known to regulate cortical layering. Positioning of DCX in this signaling pathway opens up additional possibilities of understanding how migrating neurons are controlled. PMID- 15118416 TI - hCDC4 and genetic instability in cancer. AB - About twenty years ago, scientists began to discover that colorectal cancers are caused by the sequential acquisition of genetic alterations in specific genes. To this day, we are still dissecting the genome of colorectal cancers to identify specific "culprit" genes that play a role in tumorigenesis. At the same time, we have more recently begun to turn our attention to the features of cancer cells that distinguish them from normal cells and that may be targeted therapeutically. Aneuploidy is one such hallmark of cancers, but its role in tumorigenesis is heretofore undetermined. Our efforts have focused on elucidating the fundamental mechanisms underlying aneuploidy. The assertion that a genetic basis for aneuploidy would imply its importance in tumorigenesis, and consequently make it a potential therapeutic target, represents the rationale for our pursuit of this line of research. For the last few years, we have been trying to determine whether there is a genetic cause underlying this attribute of cancers. Our recently published work entitled, "Inactivation of hCDC4 can cause chromosomal instability," attempts to address this issue and raises more questions about the cause, mechanism, timing, and therapeutic potential of genetic instability. PMID- 15118417 TI - Rambling thoughts about doing and thinking research. PMID- 15118418 TI - Nocturnal enuresis: an international evidence based management strategy. PMID- 15118419 TI - Monosymptomatic nocturnal enuresis is associated with abnormal nocturnal bladder emptying. AB - PURPOSE: We investigate the nature of enuresis episodes in monosymptomatic nocturnal enuresis using a fluid provocation model. MATERIALS AND METHODS: The study included 18 children 7 to 13 years old with monosymptomatic nocturnal enuresis. Based on basal home recordings patients were subgrouped into those with a normal nocturnal urine output and those with nocturnal polyuria (mean nocturnal urine production on wet nights exceeding 130% of functional bladder capacity, normal functional bladder capacity for age provided). Children were admitted to the hospital for 4 consecutive nights. After an adaptation night all children received orally 25 ml/kg water, 30 minutes before bedtime on the remaining 3 nights. A cordless alarm device enabled registration of enuretic episodes from another room and diapers allowed the measurement of enuresis volumes. Post-void residual volumes were measured by ultrasound. Pelvic floor electromyography was continuously recorded throughout the night, and its association to bladder emptying was investigated. RESULTS: A total of 95 enuresis and 14 nocturia episodes were recorded. Significantly more enuresis episodes were registered on nights with oral fluid load, whereas no increase in number of nocturia episodes was seen. Of the enuresis episodes 46 were associated with incomplete bladder emptying (post-void residual volume greater than 10% of total bladder volume at time of enuresis). No difference between patient groups regarding post-void residual volume was seen. Abnormal bursts of electromyography activity were associated with incomplete micturitions. CONCLUSIONS: Enuresis nocturna episodes in polyuric and nonpolyuric patients are frequently incomplete micturitions. The present findings question the definition of nocturnal enuresis episode as normal complete voiding. PMID- 15118420 TI - Sodium fraction excretion rate in nocturnal enuresis correlates with nocturnal polyuria and osmolality. AB - PURPOSE: We verify the sodium fraction excretion rate (FE Na) and potassium fraction excretion (FE K) rates in monosymptomatic nocturnal enuresis. We also correlate FE Na and FE K to urinary osmolality, nocturnal polyuria and vasopressin in the same population. MATERIALS AND METHODS: A total of 438 children 6 to 15 years old (mean age 9.7) presenting with monosymptomatic nocturnal enuresis were recruited from different centers. Inclusion criteria were 3 or greater wet nights a week, no daytime incontinence and no treatment in the previous 2 months. Exclusion criteria were cardiopathy, endocrinopathy, psychiatric problems and urinary tract abnormalities. Micturition chart, diurnal (8 am to 8 pm) and nocturnal (8 pm to 8 am) urine collection, including separate diuresis volumes, (Na, K and Ca) electrolytes and osmolality were evaluated, as well as serum electrolytes, creatinine and nocturnal (4 am) vasopressin. Diurnal and nocturnal FE K and FE Na were calculated. ANOVA test, chi-square test, Student's t test and Pearson correlation test were used for statistical analysis. RESULTS: : Nocturnal polyuria (diurnal to nocturnal diuresis ratio less than 1) was found in 273 children (62.3%, group 1 and nocturnal urine volumes were normal in 165 with enuresis (37.7%, group 2). Nocturnal FE Na was abnormal in 179 children (40.8%), including 118 in group 1 (43.2%) and 61 in group 2 (36.9%) (chi square not significant). FE Na was also increased in nocturnal versus daytime diuresis (Student's t test p <0.001). In group 1 nocturnal FE Na correlated with nocturnal diuresis (Pearson correlation p = 0.003, r = +0.175), while daytime FE Na and nocturnal FE Na correlated with diurnal diuresis (Pearson correlation p = 0.001, r = +0.225 and Pearson correlation p = 0.001, r = +0.209, respectively). In group 2 nocturnal FE Na did not correlate with diuresis (Pearson correlation p = 0.103, r = +0.128) but correlated with vasopressin values (Pearson correlation p = 0.042, r = -0.205). Urine osmolality was reduced in 140 children (31.9%) and correlated with nocturnal diuresis (Pearson correlation p = 0.003, r = -0.321). Vasopressin was decreased in 332 children (75.8%, 62.6% in group 1 and 13.2% in group 2). No significant difference was found between sexes and age of enuretic subgroups. CONCLUSIONS: Nocturnal FE Na correlates with nocturnal diuresis, whereas daytime FE Na does not. FE K in daytime and nighttime diuresis does not statistically differ in nocturnal polyuric and nonpolyuric enuretic groups. Osmolality correlates with nocturnal diuresis, and vasopressin at 4 am was lower in the nocturnal polyuric group. The hypothesis of a subset of enuretic patients presenting with nocturnal polyuria associated with high nocturnal natriuria and low vasopressin values has been confirmed. PMID- 15118421 TI - The circadian rhythm of urine production, and urinary vasopressin and prostaglandin E2 excretion in healthy children. AB - PURPOSE: In adults and adolescents the transition from day to night is followed by a pronounced decrease in diuresis, as well as reduction in the amount of osmotically active substances excreted. We investigate the circadian variations in urine production in healthy children 3 to 14 years old. MATERIALS AND METHODS: A total of 92 children completed urine collections in 2 consecutive days to be analyzed for electrolytes, urea, creatinine, osmolality, vasopressin and prostaglandin E2. RESULTS: We found a marked reduction in urine output during the night (43.41 +/- 18.53 to 25.69 +/- 12.71 ml per hour) accompanied by a decrease in the amount of electrolytes excreted (sodium 4.44 +/- 2.09 to 2.66 +/- 1.55 mmol per hour and potassium 2.38 +/-0.96 to 0.90 +/- 0.54 mmol per hour). Age and gender did not influence the observed circadian rhythm in the quantity and quality of urine production. Urinary excretion of vasopressin did not seem to reflect the circadian variations previously described for the plasma levels of the hormone. Prostaglandin E2 showed a clear circadian variation with a 30% decrease at night (32.2 +/- 19.0 to 22.0 +/- 12.6 ng/mmol creatinine). CONCLUSIONS: Healthy children exhibit pronounced circadian variations in the amount and composition of urine output with a decrease in nocturnal diuresis and excretion of osmotically active solutes. In the age range of 3 to 14 years neither age nor gender seems to affect this rhythm. Vasopressin-to-prostaglandin E2 excretion ratio appears to be of importance for regulation of urine production. PMID- 15118422 TI - Attention deficit/hyperactivity disorder in children with nocturnal enuresis. AB - PURPOSE: Although the relationship between enuresis and psychopathology has been studied intensively, little is known about the prevalence of specific psychiatric disorders. We investigate the prevalence of attention deficit/hyperactivity disorder (ADHD) in children with nocturnal enuresis and correlate these data with clinical subtypes of enuresis/incontinence. MATERIALS AND METHODS: A total of 120 children with nocturnal enuresis 6 to 12 years old participated in a prevalence study. A diagnostic interview was conducted with parents, questionnaires were completed by parents and teachers, and medical files were consulted. RESULTS: Of all enuretic children 15% were diagnosed with the full syndrome of ADHD and 22.5% met the criteria of the ADHD inattentive subtype. Data revealed that the older the children (9 to 12 years), the higher prevalence of attention deficit disorder or ADHD. Nocturnal polyuria had a significantly higher incidence in hyperactive/impulsive children but there was no significant difference in bladder function between enuretic children with or without a comorbid diagnosis of ADHD. CONCLUSIONS: The prevalence of attention deficit disorder or ADHD in nocturnal enuresis is significantly increased, especially in older children. The incidence of nocturnal polyuria is slightly increased in children who meet at least the criteria of ADHD hyperactive/impulsive subtype. No other associations between enuresis and ADHD were found. PMID- 15118423 TI - An assessment of internalizing problems in children with enuresis. AB - PURPOSE: We examine the internalizing problems (anxiety and depression) and self esteem among 9 to 12-year old children with enuresis to determine whether enuretic children are more in the clinical range, and to study the correlation between child and parent report questionnaires. MATERIALS AND METHODS: A total of 84 children with daytime and/or nighttime wetting were compared to 70 without enuresis using 5 psychometric instruments adjusted for gender and type of enuresis. Differences in mean scores, percentages of children beyond the clinical range, and correlations between child and parent report questionnaires were evaluated. RESULTS: Parental report revealed more internalizing problems ("withdrawn" and "anxious/ depressive") for children with enuresis compared to controls. A higher percentage of the study group were in the clinical range of the "total problem" scale of the Child Behavior Checklist. Child report inventories yielded no differences between groups. Moderate agreement was found between child and parent report. CONCLUSIONS: There is no evidence of internalizing problems (anxiety/depression) and low self-esteem in the self report of enuretic children. In contrast parents rate enuretic children as having more internalizing problems. Different explanations for this contradictory data are offered. Further research is necessary to explain why parents report psychological symptoms in children with enuresis. PMID- 15118424 TI - Diuretic treatment of nocturnal enuresis: preliminary results of an open pilot study. AB - PURPOSE: Since antidiuretic treatment with desmopressin in the evening has proven effective against enuresis, it would not be surprising if diuretic treatment in the afternoon would have similar effects. We are currently testing this in an open pilot investigation. MATERIALS AND METHODS: A total of 33 children including 12 girls with a mean age +/-SD of 7.6 +/- 2.3 years with monosymptomatic enuresis were recruited from an outpatient primary care setting. Wet and dry nights were recorded for 2 weeks without medication, 2 weeks with 0.4 mg desmopressin orally at night and 2 weeks with 1 mg/kg furosemide 4 to 5 hours before bedtime. RESULTS: The mean number of wet nights +/-SD during the 3 periods was 10.2 +/- 3.0, 6.7 +/- 4.7 and 7.8 +/- 4.5, respectively. The effects of desmopressin and of furosemide were statistically significant (p <0.0001 and p = 0.001), although the 2 treatments did not differ (p = 0.08). Only 6 children had a complete response (ie greater than 90% reduction in wet nights) to desmopressin and 5 responded to furosemide. Interestingly, 2 children had a clearly better response on furosemide than on desmopressin. CONCLUSIONS: The treatment of enuresis with diuretic medication in the afternoon may be beneficial but needs to be tested in randomized controlled trials. PMID- 15118425 TI - Desmopressin has an influence on the arousability of children with primary nocturnal enuresis. AB - PURPOSE: Several studies provide evidence regarding the effect of desmopressin (DDAVP) on the sleep of adults. Therefore, we investigate whether this effect has a role in children with primary nocturnal enuresis treated intranasally with DDAVP. MATERIALS AND METHODS: A prospective, randomized, placebo controlled, double-blind, crossover study was performed. Patients were assigned to 2 groups by lottery. Arousability was determined by a special bell apparatus with an adjustable sound pressure level. The wet nights per week and the results of the arousal tests were compared using the signed rank test. RESULTS: A total of 20 children with primary nocturnal enuresis 6 to 15 years old were enrolled in the study, 2 of whom had to be excluded. There were no marked differences in age or weight between the groups. The number of wet nights per week decreased significantly with DDAVP treatment. Moreover 14 patients slept more soundly with DDAVP and only 4 were more difficult to awake after the medication. This difference was significant. CONCLUSIONS: This study revealed an effect of DDAVP on arousability of enuretic children as well as its previously known action for the treatment of primary nocturnal enuresis. This result is consistent with the known action of DDAVP on sleep of elderly adults. It suggests that the cause of primary nocturnal enuresis lies in the structure of sleep of the affected patients. PMID- 15118426 TI - Ultrasound bladder measurements in patients with primary nocturnal enuresis: a urodynamic and treatment outcome correlation. AB - PURPOSE: Accurate assessment of bladder dysfunction associated with voiding dysfunctions often necessitates invasive urodynamic (UD) studies. We evaluate the use of a special ultrasound (US) protocol for the assessment of bladder dysfunction compared with urodynamic findings, and for prediction of treatment outcome in children with primary nocturnal enuresis (PNE). MATERIALS AND METHODS: US measurements were performed on 514 children 5 to 18 years old (mean age 11.2) with PNE, and compared with those of 339 normal age matched children. A US protocol was specially designed for the evaluation of bladder parameters using bladder volume and wall thickness index (BVWI %), and expected percentage bladder volume index for kidney volume. Of the enuretic children 218 had severe enuretic symptoms with more than 3 wet nights a week. They underwent urodynamic studies for detailed assessment of any underlying bladder dysfunction. A standard 4-week course of desmopressin was given to these children after the US and UD studies. The US bladder parameters were then correlated with the UD findings and treatment response to desmopressin. RESULTS: Comparing the BVWI in normal and enuretic children in correlation with functional bladder capacities we were able to delineate bladder wall thickness and capacity as BVWI less than 70-small capacity bladder with thick wall, BVWI 70 to 130-normal bladder capacity with normal wall thickness and BVWI greater than 130-large bladder capacity with thin wall. There were statistically significant correlations between BVWI and treatment response. In addition, there was a high predictive value of normal bladder function with a normal BVWI. Patients with good response to treatment had normal BVWI, whereas poor response to treatment was significantly associated with pathological bladder conditions, that is small bladder capacity with thick bladder wall or large bladder capacity with thin bladder wall (p <0.0001). Of note, abnormalities detected by UD correlated well with bladder abnormalities measured by US. CONCLUSIONS: PNE comprises a diverse spectrum of conditions resulting in a mismatch of nocturnal urine production in excess of nocturnal functional bladder capacity, and underlying bladder dysfunction has an important role in the pathophysiology especially in refractory cases. This US protocol can provide useful predictive clues, which may be helpful to differentiate treatment subtypes, guide clinical management and minimize the need for invasive urodynamic studies. PMID- 15118427 TI - Urodynamic findings in adults with primary nocturnal enuresis. AB - PURPOSE: We evaluate bladder function in adults with primary nocturnal enuresis (PNE) since childhood. MATERIALS AND METHODS: We recruited all patients older than 16 years who presented to our PNE clinic with persistent enuretic symptoms. All patients were assessed by a detailed voiding diary, uroflowmetry and ultrasonography of the urinary tract. Urodynamic studies were performed for those with moderate or severe PNE (more than 3 wet nights a week). RESULTS: The study included 18 males and 29 females with a mean age of 20 years (range 16 to 43). Of these patients 37 (79%) had moderate or severe symptoms and 17 (38%) also had daytime urinary symptoms. Urodynamic studies were conducted in 30 patients, including 12 males and 16 females (93%) with detrusor overactivity. In addition, 73% of patients had urodynamic evidence of functional bladder outflow obstruction, including dysfunctional voiding and detrusor sphincter or detrusor pelvic discoordination. Two male patients (6.7%) had an obstructive pattern on urodynamics and subsequent cystoscopic examination confirmed the presence of congenital obstructive urethral lesions. Sixteen patients (53%) had significantly reduced bladder capacity of less than 300 ml. CONCLUSIONS: Unlike in early childhood, the majority (79%) of enuretic adults had significant symptoms, and more than a third also had daytime urinary symptoms. We believe that PNE in adults is not only a psychologically disturbing condition, but also a urological disorder with significant underlying bladder dysfunction that warrants special attention to management. PMID- 15118428 TI - Long-term efficacy and predictive factors of full spectrum therapy for nocturnal enuresis. AB - PURPOSE: We determine the effect and predictive factors of relapse 1 year after combination therapy of an enuresis alarm, bladder training, motivational therapy and retention control training for nocturnal enuresis. MATERIALS AND METHODS: In 68 of 77 children the relapse rate 1 year after full spectrum therapy for nocturnal enuresis was investigated. Gender, age, sleep arousal, family history, monosymptomatic, bladder capacity, overactive bladder, nighttime polyuria, duration of treatment, over learning and psychosocial factors were investigated. RESULTS: The relapse rate during the whole year was 50%, with 33.8% of subjects being dry and 16.2% sometimes wet. The relapse rate after 1 year was 16%. Nine patients could not be reached and, thus, were considered dropouts. Only overactive bladder and psychosocial problems were significantly related to relapse. CONCLUSIONS: The relapse rate during the year was high while the relapse rate after 1 year was low. Psychosocial problems and overactive bladder were the only 2 predictive factors for relapse. PMID- 15118429 TI - Intermittent oral desmopressin therapy for monosymptomatic primary nocturnal enuresis. AB - PURPOSE: In a prospective study we evaluate the efficacy of intermittent desmopressin (DDAVP) every other day for patients with nocturnal enuresis relapse who need additional therapy. MATERIALS AND METHODS: Between January 2000 and August 2001, 71 boys and 52 girls 6 to 22 years old (mean age 12.5) were treated with 0.2 mg DDAVP daily for monosymptomatic nocturnal enuresis. After an initial 2 weeks of dose titration the nonresponders were given 0.4 mg DDAVP daily. Those who did not respond to this dose were excluded from study. The remaining patients took desmopressin for 3 months. Patients with persistent enuresis after 3 months of treatment were given intermittent 0.2 or 0.4 mg DDAVP every other day. Followup was performed 6 weeks later. RESULTS: Of 123 patients 92 completed the study. Mean followup after beginning intermittent DDAVP therapy was 9.2 months (range 6 to 18). Of the 92 patients 45 responded to the 0.2 mg daily dose (group 1) and continued treatment for at least 3 months, while the dose was titrated to 0.4 mg for the remaining 47 (group 2). There were 23 patients who did not respond to 0.4 mg DDAVP and they were excluded from the study. After cessation of the drug 21 group 1 patients (46.6%) and 13 group 2 patients (54%) still had enuresis, and they were placed on intermittent therapy. After 6 weeks 15 of these 34 patients had complete and 13 of the remaining 19 had partial response, while the 6 nonresponders continued on daily DDAVP. Overall the complete and partial response rate of intermittent treatment was 20 of 21 group 1 patients (95%) and 8 of 13 group 2 (61.5%). CONCLUSIONS: For some enuretic patients with relapse after cessation of initial 3-month therapy, intermittent DDAVP may be an effective alternative long-term treatment. PMID- 15118430 TI - Imipramine for therapy resistant enuresis: a retrospective evaluation. AB - PURPOSE: Imipramine has a proven effect in enuresis but is now seldom used. We reviewed the records of children who had been treated with imipramine to determine prognostic indicators. MATERIALS AND METHODS: The records of all enuretic children treated with imipramine at our clinic were retrospectively evaluated, with special emphasis on history, and for most children bladder volume, renal concentrating capacity and urine production. RESULTS: All 49 children included in the study had previously received desmopressin, alarm and anticholinergic treatment without success. Of the children 31 (64.6%) were responders (R), with at least 50% reduction in enuresis frequency (22 became completely dry) and 17 (35.4%) were nonresponders (NR). Older age (R 11.4 +/- 3.4, NR 8.7 +/-1.8 years, p = 0.004) and low spontaneous bladder capacity (R 2.6 +/- 0.9, NR 3.4 +/- 0.9 ml/kg body weight, p = 0.03) were prognostically favorable, whereas constipation (p = 0.02) and a history of daytime incontinence (p = 0.04) indicated poor prognosis. Ten children experienced nausea or other minor problems. Seven children with attention deficit and hyperactivity became more focused during treatment. CONCLUSIONS: Imipramine is a useful treatment for enuresis when everything else has failed, especially among older children. Factors related to detrusor function provide prognostic information. PMID- 15118432 TI - Enuresis alarm treatment as a second line to pharmacotherapy in children with monosymptomatic nocturnal enuresis. AB - PURPOSE: We evaluate the effectiveness of enuresis alarm as second line therapy for partial or nonresponders to pharmacotherapy. MATERIALS AND METHODS: We recommended enuresis alarm treatment for 67 partial or nonresponders to pharmacotherapy for monosymptomatic nocturnal enuresis. Of these patients 28 were evaluable by inclusion and exclusion criteria. We analyzed the effectiveness of enuresis alarm therapy using the response criteria defined by the reduction rate of wet nights, defined as complete response (greater than 90%), partial response (50% to 90%) and no response (less than 50%). We defined initial and lasting cure when patients showed persistent full response for 4 weeks and 6 months after cessation of all treatments, respectively. RESULTS: After pharmacotherapy partial responders showed a mean response of 81.2%. On the other hand, nonresponders exhibited a mean response of 26.4% with more than 20 wet nights in 4 weeks. After using second line enuresis alarm treatment 90.5% (19 of 21) of partial responders became full responders and 71.4% (15 of 21), 61.9% (13 of 21) of partial responders showed initial and lasting cure, respectively. In addition, 71.4% (5 of 7) of nonresponders became full responders and 57.1% (4 of 7) exhibited initial and lasting cure. CONCLUSIONS: Enuresis alarm as second line therapy for monosymptomatic nocturnal enuresis is effective for lasting cure as well as high initial full response rate in partial and nonresponders to pharmacotherapy. Therefore, enuresis alarm is a reasonable second line therapeutic option for partial or nonresponders to pharmacotherapy. PMID- 15118431 TI - The effect of alarm treatment on the functional bladder capacity in children with monosymptomatic nocturnal enuresis. AB - PURPOSE: We studied the characteristics of a group of monosymptomatic nocturnal enuretics successfully treated with the alarm system, with special reference to changes in functional bladder capacity. MATERIALS AND METHODS: The diaries of 7 girls and 19 boys 7 to 13 years old with severe nocturnal enuresis, small daytime bladder capacity (70% or less of expected capacity for age) and poor or absent response to desmopressin were analyzed. Patients were treated with an alarm until complete dryness was achieved for 21 consecutive nights before ending therapy. Immediately after the treatment they recorded a 1-week followup diary of voiding and fluid intake. RESULTS: Mean duration of the alarm treatment was 82 days, and there was no change in nocturnal or 24-hour diuresis from baseline to followup. Nocturia developed during the alarm treatment in 48% of the children. The nocturnal diuresis on nocturia nights was significantly higher than on nights without nocturia. Daytime functional bladder capacity increased significantly in children with and without nocturia. CONCLUSIONS: Treatment with an alarm system increases daytime functional bladder capacity significantly in children with and without nocturia. A higher nocturnal urine production on nocturia nights explains why some children have nocturia and others do not. PMID- 15118433 TI - Increased urinary calcium excretion in enuretic children treated with desmopressin. AB - PURPOSE: The use of desmopressin in the treatment of primary nocturnal enuresis (PNE) is accepted and based on the fact that this drug leads to renal water reabsorption. However, recent findings have also implicated that desmopressin regulates other molecules, such as sodium and potassium. We investigate if desmopressin influences renal Ca2+ handling. MATERIALS AND METHODS: A total of 32 children with PNE were enrolled in a prospective study. Patients received a standard 30 microg desmopressin intranasally before going to bed. All patients were treated for at least 4 weeks. Desmopressin was then withdrawn and reintroduced after 2 weeks. Urine samples were collected during all 3 phases of the study. Ca2+ measurement was performed in single morning spot urines as well as in 24-hour collections. Additionally, blood was sampled for analysis of Ca2+. The Wilcoxon signed rank test was used for statistical analysis. RESULTS: Wet nights decreased an average of 4.75 to 1.0 per week with desmopressin treatment. While blood concentrations did not change with or without medication, urinary Ca2+ excretion was significantly higher while patients were treated with desmopressin. This significant result was the same in single spot as well as in 24-hour samples. CONCLUSIONS: This study demonstrated the increased excretion of Ca2+ by desmopressin treatment in children with PNE. Since Ca2+ is a crucial molecule in growth and development, this finding indicates the necessity of larger followup studies concerning Ca2+ handling and growth in children on long term desmopressin treatment. PMID- 15118434 TI - Is prophylactic antimicrobial treatment necessary after hypospadias repair? AB - PURPOSE: We evaluate the complication rate after hypospadias repair with and without the use of antimicrobial prophylaxis. MATERIALS AND METHODS: A total of 101 boys who underwent tubularized incised plate urethroplasty with urethral catheter placement during a 16-month period were randomly divided into group 1-52 treated with cephalexin from day 1 after surgery to 2 days after catheter removal and group 2-49 who did not receive prophylaxis. All children received cefonicid before surgery. RESULTS: Average patient age was 2.3 years (range 11 months to 6.5 years). Hypospadias was coronal in 54 boys, penile in 33, glanular in 9 and penoscrotal in 5, the distribution of which was similar in both groups. Median time to urethral catheter removal was 8.6 days in group 1 and 8.3 in group 2. Overall bacteriuria was noted in 11 children in group 1 and 25 in group 2. The most common pathogen was Pseudomonas aeruginosa in group 1 and Klebsiella pneumoniae in group 2. Urethrocutaneous fistula developed in 3 boys in group 1 and 9 in group 2, meatal stenosis occurred in 1 boy in group 1 and 4 in group 2, and 1 boy in group 1 had meatal regression. Three boys in group 1 and 12 in group 2 had a complicated urinary tract infection (p <0.05). There was no difference in the number of surgical complications between boys for whom this was the first operation or a repeat hypospadias repair. CONCLUSIONS: A broad-spectrum antibiotic is recommended before and antimicrobial prophylaxis after hypospadias repair. This protocol may decrease the risk of complicated urinary tract infections after surgery, and probably reduce meatal stenosis and urethrocutaneous rates. PMID- 15118435 TI - Do patch procedures prevent complications of the Mathieu technique? AB - PURPOSE: The effects of V-incision sutured (MAVIS) and dartos patch reinforcement techniques in reducing side effects and complications of the Mathieu procedure and improving the appearance of the meatus are evaluated. MATERIALS AND METHODS: A total of 74 patients with distal shaft hypospadias without chordee or with minimal chordee were operated on by a single surgeon using Mathieu's technique. While only the classic Mathieu repair was performed in group 1, MAVIS and dorsal dartos patch reinforcement was included with the procedure in group 2. A silicone urethral catheter and foam silicone dressing were used in both patient groups. RESULTS: There were 32 cases in group 1 and 42 in group 2. Median patient age at operation was 3.2 and 3.4 years in groups 1 and 2, respectively. Mean postoperative period for the silicone catheter was 4.9 days in both groups. Followup ranged from 52 to 128 months (mean 76) for group 1 and 13 to 43 months (25.4) for group 2. Five fistulas and 1 meatal stricture developed in group 1 and all the children in this group required meatal calibration with dilatation in some. Neither fistula nor stricture occurred in group 2 and there was no need for calibration. A slit-like meatus was achieved in all children in group 2 but in only 12 in group 1. CONCLUSIONS: The MAVIS technique and dorsal dartos patch reinforcement may prevent complications of the classic Mathieu procedure. A slit like meatus was obtained in all cases with this approach. PMID- 15118436 TI - Periurethral constrictor in pediatric urology: long-term followup. AB - PURPOSE: We report long-term followup of a new device for the treatment of urinary incontinence in children. MATERIALS AND METHODS: A periurethral constrictor was implanted in 29 boys and 13 girls 3 to 17 years old (mean age 10.2, median age 10) during the last 9 years. Of the patients 29 had neurogenic bladder, 12 had bladder exstrophy and 1 had megalourethra. Bladder augmentation was done simultaneously in 34 patients. The device was implanted around the bladder neck in 41 cases and at the bulbous urethra in 1. Continence was achieved in all cases when the device remained in situ. RESULTS: Followup ranged from 4 to 104 months (mean 63, median 75). In 23 patients (82.1%) in the neurogenic group and the patient with megalourethra the device remains in situ and continence is preserved. Clean intermittent catheterization is performed without difficulty. In 4 patients the device was extracted due to erosion or infection. The device was extracted due to erosion and urine extravasation in 10 exstrophy group patients. Two patients have the device and perform post-void catheterization. CONCLUSIONS: Based on the data from this long-term study we conclude that the periurethral constrictor is a safe alternative for the treatment of urinary incontinence in children. Like other devices it must be used with caution in cases of bladder exstrophy. PMID- 15118437 TI - Computer assisted pyeloplasty in children: the retroperitoneal approach. AB - PURPOSE: We describe the first series of computer assisted retroperitoneoscopic pyeloplasty in children using the Da Vinci Surgical System (Intuitive Surgical, Inc., Mountainview, California) with regard to setup, method, operation time, complications and preliminary outcome. The small space in the retroperitoneum of children and larger instruments of the computer assisted surgical system make modification of the retroperitoneal access necessary. MATERIALS AND METHODS: In 13 children with a median age of 6.7 years (range 3.5 to 16.2) and ureteropelvic junction obstruction 15 pyeloplasties were performed with the Da Vinci Surgical System. With the patient in a lateral semiprone position the retroperitoneal space was developed by blunt and balloon dissection. Three ports were placed for the computer assisted system and 1 for assistance. Pyeloplasty was performed with the mounted system placed behind the patient. RESULTS: The procedures were completed in all patients with the computer assisted system. Median operative time was 173 minutes (range 76 to 215) and there were no perioperative complications. Median postoperative hospital stay was 2 days (range 1 to 3). Two patients had postoperative complications related to the Double-J catheter (Cook Urological Inc., Spencer, Indiana). In 1 patient the catheter was displaced with its lower end in the distal ureter, and the other patient was rehospitalized with occlusion of the catheter and treated with nephrostomy for a few days. All patients had a satisfying outcome during the preliminary followup period of 1 to 7 months. CONCLUSIONS: In this the first series of computer-assisted retroperitoneal pyeloplasty in children the method seems feasible with shorter operative time and similar complications as in standard retroperitoneoscopic procedures but easier handling of the instruments and shorter training for the surgeon. The costs for the system are still high and only longer followup will reveal whether the more precise placement of sutures and the advantage of the magnified 3-dimensional view lead to at least the same results as the open procedure. PMID- 15118438 TI - Laparoscopic vaginal reconstruction using a sigmoid colon segment: a preliminary report. AB - PURPOSE: Sigmoid vaginoplasty is an alternative technique for vaginal replacement. We describe the successful use of laparoscopy for reconstruction of a vagina in patients with the Mayer-Rokitansky-Kuster-Hauser syndrome. MATERIALS AND METHODS: Laparoscopic surgery was performed in 5 girls with a mean age of 17 years and 5 months. We used 4 ports, the first of which was inserted supraumbilically by the Hasson technique. The next 10 mm port was introduced through the right lower quadrant under visual control. Then 5 and 15 mm ports were inserted at the left lower quadrant. The scope was introduced through the supraumbilical port. After isolating a segment of the sigmoid using 2 endoscopic staplers an incision was made in the deepest part of the vestibule. Performing a blunt dissection a 2-finger wide space was created between the urethra, bladder and rectum. In the top of the space the peritoneum was incised under laparoscopic control, which allowed passage of the forceps from the perineum and enabled descent of the isolated sigmoid segment. The graft was fixed to the mucosa of the vaginal vestibule using a single suture. The continuity of the intestinal tract was restored using a circular mechanical suture through the rectum. RESULTS: Average operative time was 5 hours. None of the cases needed conversion to open surgery. No significant blood loss or any other complication was noted. Average postoperative stay was 7 to 9 days. CONCLUSIONS: This preliminary report suggests that laparoscopic sigmoid vaginal replacement is feasible and safe, and may be used as an alternative to open surgery. PMID- 15118439 TI - Nonsecretory intestinocystoplasty: a 10-year experience. AB - PURPOSE: We present long-term results on the use of demucosalized intestine for reconstructive surgery of the bladder. MATERIALS AND METHODS: A total of 129 bladder augmentations with demucosalized intestine were performed in 123 patients (55% males and 45% females) 3 months to 53 years old during the last 10 years. Of the patients 82 presented with neurogenic bladder, 40 with bladder exstrophy, 3 each with tuberculosis and posterior urethral valves, and 1 with female hypospadias. Sigmoid was used in 104 cases and ileum in 25. In 105 cases a silicone balloon was left inside the augmented bladder for 2 weeks. In the remaining cases bladder mucosa was preserved and no mold was used. A silicone inflatable bladder neck cuff was implanted at the same time as augmentation in 32 patients. RESULTS: Followup ranged from 3 to 135 months (median 51.0). There was a 329% increase in bladder capacity and compliance increased 7-fold. There were 13 (10.1%) cases considered failures, which were treated with reaugmentation using demucosalized ileum (6), different forms of augmentation (5) and no reaugmentation (2). There was no mucus formation, bladder perforation or neoplasia. CONCLUSIONS: Based on our study we conclude that demucosalized intestine is a safe alternative for bladder augmentation and can be used for the same indications as total bowel segments. PMID- 15118440 TI - Functional constipation in children. AB - PURPOSE: Constipation in children increases the likelihood of urinary incontinence, bladder overactivity, dyscoordinated voiding, a large capacity, poorly emptying bladder, recurrent urinary tract infection and deterioration of vesicoureteral reflux. We present a consensus related to the assessment, diagnosis and treatment of children with bowel dysfunction coexisting with a known disorder of urinary continence or voiding coordination. MATERIALS AND METHODS: A panel of international multidisciplinary clinicians working on pediatric continence care was invited to participate in the First International Children's Continence Society Bowel Dysfunction Workshop. The seminar sought to address the interrelationship of bowel dysfunction with disorders of urinary continence or voiding mechanics. RESULTS: Constipation is an end point defined by a constellation of symptoms, including infrequent passage of stool, difficulty passing stool, feces that are either large and hard or in small pieces, abdominal pain, palpable stool in the abdomen, stool in the rectal vault, loading on x-ray or fecal soiling. Assessment was done to identify potential organic causes of constipation, clarify symptoms, and identify altered motor behavior and abdomino/pelvic floor muscle incoordination. Whether the underlying problem was one of stool consistency, poor cognition, motivation or fear on the part of the child, or whether it related to gut motility, rectal sensation, stool retention or disordered emptying mechanics, the definitive therapy begins with rectal emptying of impacted stool followed by maintenance of regular soft stools to eliminate fear of pain with defecation. CONCLUSIONS: Constipation is a challenge to the clinician but with comprehensive assessment and systematic intervention children can achieve independent bowel emptying, which positively impacts bladder function. PMID- 15118441 TI - Comorbidity of functional urinary incontinence and encopresis: somatic and behavioral associations. AB - PURPOSE: Functional urinary incontinence and encopresis are common comorbid disorders in childhood. We analyze the specific somatic and behavioral symptoms associated with functional enuresis/urinary incontinence and encopresis when they occur together. MATERIALS AND METHODS: A total of 167 consecutive children 5 to 10 year olds, with day and/or night wetting were examined prospectively with ultrasound, uroflowmetry, electroencephalography, the Child Behavior Checklist, Culture Fair Intelligence Test and ICD-10 child psychiatric diagnoses. RESULTS: The main findings for the comorbid group (20 patients) with wetting and soiling were a significantly higher rate of daytime incontinence and micturition problems, thickened bladder walls and pathological electroencephalography. There were higher, although not significant, rates of previous urinary tract infections, antibiotic prophylaxis, residual volume and abnormal uroflow curves in this group. Behaviorally, hyperkinetic syndromes, and emotional and conduct disorders (according to ICD-10) were more common. Of the 20 patients 65% had a Child Behavior Checklist total score (greater than 90th percentile) in the clinical range. The externalizing, internalizing, delinquent and anxious/depressed problem scales were also significantly higher. CONCLUSIONS: This risk group requires detailed assessment and specific treatment. In addition to the symptomatic treatment of the wetting and soiling, many of these children are in need of specific behavioral, psychotherapeutic and pharmacological treatment. PMID- 15118442 TI - Intravesical electrical stimulation improves neurogenic bowel dysfunction in children with spina bifida. AB - PURPOSE: We observed concomitant improvement in fecal incontinence in children with myelomeningocele undergoing intravesical electrical stimulation (IVES) to decrease uninhibited bladder contractions and increase bladder capacity and/or bladder sensation. We retrospectively reviewed the effect of intravesical electrical stimulation on neurogenic bowel dysfunction in these children. MATERIALS AND METHODS: A total of 9 boys and 15 girls 3.9 to 13.2 years old (mean age 8.1) completed a mean of 30.3 daily sessions (range 10 to 69) of IVES. Evaluation forms were used to record frequency of fecal incontinence, daily bowel movement and diaper use before and after IVES. RESULTS: The mean number of overall fecal incontinence episodes decreased significantly from 7.36 to 4.8 a week after IVES (p <0.05). Greater than 50% decrease in the episodes of fecal incontinence was observed in 75% of the patients. However, there was no significant change in the number of daily bowel movements before (1.8 daily) and after (1.55 daily) IVES. CONCLUSIONS: These results demonstrate the therapeutic effect of IVES in children with neurogenic bowel dysfunction and spina bifida. We believe that IVES is another viable option for controlling fecal incontinence in these children. PMID- 15118443 TI - Voiding dysfunction in x-linked adrenoleukodystrophy: symptom score and urodynamic findings. AB - PURPOSE: Adrenoleukodystrophy is an x-linked genetic disorder characterized by the breakdown of the myelin sheath surrounding nerve cells in the brain and progressive dysfunction of the adrenal gland with 6 possible phenotypes. Lower urinary tract symptoms affect the quality of life of these patients but only sporadic cases have been previously studied by urodynamics. We report to our knowledge the first series in the literature looking at the prevalent pattern. MATERIALS AND METHODS: A total of 14 children and adults 8 to 53 years old (average age 30) with different phenotypes were referred for evaluation. Of the patients 6 had adrenomyeloneuropathy without cerebral involvement (C-) 3 had the adult cerebral form, 3 had the childhood cerebral form, 1 had cerebral involvement (C+) and 1 was asymptomatic. All patients were submitted to a screening protocol, consisting of the self-administered short form questionnaire International Conference on Incontinence Questionnaire-Short Form (ICIQ-SF) and a 3-day voiding diary. Patients were divided in group 1-no urinary symptoms, ICIQ SF score 0 and no further investigation (3); group 2-frequency/urgency, ICIQ-SF score 1 to 5, uroflowmetry and post-voiding residual urine (4); and group 3 voiding symptoms and incontinence, ICIQ-SF score greater than 5, standard urodynamic examination (7). RESULTS: A neurogenic overactive bladder was the prevalent urodynamic finding (6 of 7 patients), while hypocontractility was found in 1 case. CONCLUSIONS: Vesicosphincteric dysfunction can be explained by spinal and/or cerebral involvement in the different neurological forms of adrenoleukodystrophy. Attempts at treatment with anticholinergics and neuromodulation in 4 of our patients provided temporary benefit with clear subjective amelioration of symptoms and improved quality of life. PMID- 15118444 TI - The outcome of detrusor myotomy in children with neurogenic bladder dysfunction. AB - PURPOSE: We evaluate the outcome of detrusor myotomy for neurogenic bladder dysfunction (NBD) in children. MATERIALS AND METHODS: A retrospective analysis was performed of data compiled from medical and urodynamic records of children younger than 16 years with NBD who had undergone detrusor myotomy from 1992 to 2000 at our department. RESULTS: Surgery was performed in 14 children with a mean age +/- SD of 6.7 +/- 4.3 years (range 0.9 to 14.2) and mean followup of 5.9 +/- 1.7 years. All patients were diagnosed with NBD, which was the result of myelomeningocele in 9, sacral agenesis in 2, lumbosacral lipoma in 1, multiple vertebral anomalies in 1 and spinal neuroblastoma in 1. Main indications for surgery included urinary incontinence in 11 cases and high pressure/low capacity bladders with vesicoureteral reflux and impending renal damage in 8. No major postoperative complications were recorded. Although mean maximal cystometric bladder capacity was unchanged 1 month postoperatively (89.7 +/- 70.6 ml) compared to preoperatively (92.5 +/- 75.1 ml, p = 0.87), significant increments of 216%, 237% and 292% were measurable at 3 months, 1 year and 5 years, respectively. Ultimately most of the patients approached age specific capacities. Complete continence on clean intermittent catheterization was achieved by 8 of 11 patients and improved markedly in 1. Reflux was alleviated in 6 cases and improved in 1. Kidney function developed normally in all but 1 patient with persistent reflux. CONCLUSIONS: When feasible, detrusor myotomy offers a safe and effective alternative for the management of pharmacologically intractable NBD in children. PMID- 15118445 TI - Variability in normative urine flow rates. AB - PURPOSE: Parameters derived from uroflowmetry are frequently used in the evaluation and reassessment of children presenting with lower urinary tract dysfunction. Since current nomograms have been constructed from 1 to 2 voids per child, variability of flow parameters is unknown. We evaluate intraindividual variability of flow parameters in children free of lower urinary tract symptoms. MATERIALS AND METHODS: Children between 4 and 16 years old (mean age 9.8) who were hospitalized but not suffering from urinary tract infection, pyelonephritis or neurological disturbance, and who had neither structural abnormality of the urinary tract nor acute pain or disorientation voided spontaneously over a flowmeter on 4 to 6 occasions. Post-void urine was estimated by ultrasound within 5 minutes of micturition to confirm complete emptying. RESULTS: A total of 98 subjects provided a mean of 4.4 traces each. Neither maximum nor average flow rate showed great intraindividual variability (maximum flow rate r >0.8, average flow rate r >0.74). Flow rates significantly correlated with flow duration, volume voided and patient age. Initial flow curves were bell-shaped in 63% of cases, staccato in 30% and intermittent in 6%. These proportions did not change with subsequent voids. There was no significant difference between genders with respect to staccato voiding, although boys demonstrated 70% of intermittent voids and were significantly older than girls. CONCLUSIONS: In normal children there is minimal variability in flow rates and the phenomenon of staccato voiding is seen approximately 30% of the time, despite no significant levels of post-void residual urine. PMID- 15118446 TI - Male fetal pig lower urinary tract function. Part II: free voiding pattern close to term and in the newborn. AB - PURPOSE: We assessed the development of natural voiding function late in gestation and in the immediate postnatal period in a porcine model. MATERIALS AND METHODS: The study comprised 7 male fetal minipigs (median age 94 days, 0.88 gestation) and 7 male newborn pigs. In all pigs an ultrasonic probe was placed around the subcutaneous urethra and pressure catheters were placed in the bladder and rectum and in the amnionic cavity in the fetal pigs. The catheters were connected to pressure transducers as the flow probe was connected to a flow meter, and the recordings were sampled at a rate of 10 Hz on a personal computer. RESULTS: The newborns had a median voiding frequency of 3.3 times per hour while the fetuses voided a median of 5.9 times per hour (p = 0.16). Both groups voided with a staccato flow at a frequency of 1 to 2 Hz, indicating voiding dyscoordination between the detrusor and urethra. Maximum flow rate increased from median 4.4 ml per minute (range 2.0 to 8.8) in the fetal group to median 10.9 (5.4 to 18.3) in the newborns (p = 0.07). While the minimum opening pressure remained unchanged (median 7.0 cm H2O, range 5.7 to 13.0) vs median 7.6, (range 7.7 to 14.0, p = 0.57) the detrusor pressure at maximum flow decreased from median 22.4 cm H2O (range 16.6 to 39.0) in the fetal pigs to 12.1 cm H2O (8.3 to 22.3) in the newborns. CONCLUSIONS: Fetal and newborn pigs have dyscordinated voiding with a staccato flow. While the urethral opening pressure appears to be unchanged, the detrusor pressure at maximum flow decreases during the last period of gestation, indicating decreased urethral resistance. These findings are in accordance with observations made in human infants. PMID- 15118447 TI - Is spina bifida occulta associated with lower urinary tract dysfunction in children? AB - PURPOSE: Of congenital malformations of the central nervous system 46% are abnormalities of the spinal cord, which includes spina bifida occulta. The occurrence and significance of spina bifida occulta in children with lower urinary tract and bowel dysfunction were evaluated prospectively. MATERIALS AND METHODS: Between January 1999 and February 2001, 158 consecutive children presenting with lower urinary tract and bowel dysfunction were prospectively evaluated for associated spinal abnormalities. Plain abdominal x-ray showed either the presence or absence of fusion of the posterior elements of the lumbar and/or sacral spinous processes. Of the 158 patients 57 (36%) had spina bifida occulta (group 1) and 101 (64%) did not (group 2). RESULTS: There was no correlation between clinical outcome in group 1 with spina bifida occulta and lower urinary tract dysfunction and the pattern of spina bifida occulta. Significant correlation was present between lower urinary tract dysfunction and the presence of spina bifida occulta on plain abdominal x-ray. No significant correlation was found between the presence of spina bifida occulta and spinal cord abnormalities by magnetic resonance imaging scan. There was no direct causal relation between the radiological finding and lower urinary tract dysfunction. CONCLUSIONS: Children presenting with spina bifida occulta, and lower urinary tract and bowel dysfunction have normal genitourinary tracts and absent spinal cord abnormalities. Spina bifida occulta is probably a coincidental finding and its true significance in this cohort was not established. PMID- 15118448 TI - An empirical treatment algorithm for incontinent children. AB - PURPOSE: Successful response rates of monotherapeutic strategies in urge incontinent children are limited. We evaluate whether adjuvant treatment improves outcomes. MATERIALS AND METHODS: Incontinent children were evaluated according to International Consultation on Incontinence standards. Propiverine (0.4 mg/kg) was applied 2 times daily for 4 weeks (treatment period 1) before reevaluation. Primary outcome was achievement of continence and secondary outcome was improvement of functional bladder capacity. In partial responders an alternative adjuvant treatment was initiated for another 12 weeks (treatment period 2). RESULTS: Of 70 enrolled patients 29 achieved continence (responders) and 35 responded partially and were assigned to adjuvant treatment, which consisted of selective alpha-blocker for functional bladder outflow obstruction (6), desmopressin for excessive nocturnal urine production (19) and biofeedback for increased pelvic floor activity during micturition (10). Only 6 nonresponders (9%) were assigned to specialized management. After treatment 2, 20 of the 35 partial responders achieved continence, thus avoiding specialized management. CONCLUSIONS: Propiverine monotherapy for incontinent children is effective. However, applying adjuvant treatment modalities to partial responders increases overall efficacy rates. PMID- 15118449 TI - Review of panel discussions. PMID- 15118452 TI - Infusion therapy: a multifaceted approach to teaching in nursing. AB - A multifaceted method of instruction for infusion therapy is presented in this article. The traditional use of mannequin arms is included and virtual reality is incorporated. The union of these methods, both of which use a hands-on approach, complements the use of text and laboratory demonstration for teaching infusion therapy. This study shows that with increased use of the virtual reality system, the student has increased probability of success at the first attempt during the return demonstration of the skill of initiating infusion therapy. With the emphasis on enhancing student preparedness for this advanced skill, the authors have found that the inclusion of all methods will better meet students' learning needs and their preferred methods of learning. PMID- 15118453 TI - Contamination control in nursing with filtration: part 2: emerging rationale for bedside (final) filtration of prestorage leukocyte-reduced blood products. AB - The first part of this 2-part series focused on the manufacture of filters and the application of filtration technology to intravenous fluids and point-of-care hospital water. This second part describes an apparent emerging potential for final filtration defined as bedside filtration of blood and component blood products leukocyte-reduced at the blood center prior to storage. Final filtration serves to further reduce the leukocyte burden in a previously leukocyte-reduced blood product. Another target for final filtration includes putative soluble mediators of morbidity.Selected patients may be at greater risk for alloimmunization and refractory to the benefits afforded by transfusion of blood leukocyte reduced to the current established standards. Multiparous patients who subsequently find themselves in need of a transplanted organ are alloimmunized by exposure to fetal proteins and may be further alloimmunized by transfusion. Such effects can put them at risk for increased latency for donor organ availability and organ rejection. Kidney transplant patients find themselves the recipients of transfused blood products particularly during end-stage renal disease and recent data suggest such patients are not benefited by the levels of leukoreduction prescribed by current standards and may need more dramatic leukocyte removal. The process of blood production is described and affords a greater appreciation for the levels of white cells found in component blood products. The development of alloimmunization is reviewed and fosters greater appreciation for a discussion of the potential for therapeutic value of more dramatic leukocyte reduction and blood conditioning accomplished through the removal of soluble mediators of morbidity. PMID- 15118454 TI - Pediatric parenteral nutrition: differences in practice from adult care. AB - Pediatric and adult parenteral nutrition have more similarities than differences. The differences between the two practices become apparent in the time frame for starting support, a few pediatric-specific products, and growth monitoring. The biggest challenge facing the practitioner is to familiarize him- or herself with the wide range of requirements for macro- and micronutrients. PMID- 15118455 TI - The use of alteplase for restoring patency to occluded central venous access devices in infants and children. AB - A 21-month retrospective review was completed at the Lucile Packard Children's Hospital to assess the experience of 22 infants and children who received alteplase for the clearance of occluded central venous access devices. After the first dose, 86% (n = 19) of the catheters cleared. Two additional catheters cleared with a second dose. With alteplase treatment, 95% (n = 21) of the catheters cleared. No adverse events were noted within 24 hours after the alteplase was received. Infusion of alteplase appeared to be safe and effective in restoring patency to occluded central venous access devices in infants and children. PMID- 15118456 TI - Incidence and nature of epidemic nosocomial infections. AB - Nosocomial bloodstream infections continue to be a major cause of morbidity and mortality. Approximately 8% of all nosocomial infections reported in the United States are primarily bloodstream infections. These infections prolong hospital length of stay, increase mortality, and raise the overall cost of healthcare. A contaminated infusate administered through a central venous catheter is one of the commonly identified causes of nosocomial bacteremia. In most cases, contamination of the infusate occurs extrinsically during manipulation of the fluid before its administration to the patient. Failure to use aseptic technique and poor hand washing often are the cause. In addition to improved staff education, surveillance for nosocomial bloodstream infections continues to be the cornerstone of prevention. PMID- 15118457 TI - Peripherally inserted central catheter malposition in a persistent left superior vena cava. AB - Persistent left superior vena cava (PLSVC) is the most common congenital anomaly of the superior vena cava system. Left superior vena cava exists in early embryonic development, but this vessel degenerates as the cardiovascular system matures. Failure of this process to occur results in PLSVC. If not associated with other cardiac malformations, PLSVC has no clinical signs or symptoms. This article discusses a case of PLSVC diagnosed after placement of two peripherally inserted central venous catheters that appeared to malposition outside the central circulation. After unsuccessful bedside placement of the two catheters, the patient was referred to interventional radiology for placement of a Hickman catheter. A contrast injection study showed the presence of a PLSVC with the absence of a normal right superior vena cava. The tip of the peripherally inserted central venous catheter was found to be at a satisfactory location within the PLSVC. PMID- 15118459 TI - Cumulative incidence of converting from clinically unilateral to bilateral exfoliation syndrome. AB - PURPOSE: To obtain long-term cumulative incidence estimates of time to conversion of clinically unilateral to bilateral exfoliation syndrome (ES). METHODS: A retrospective, longitudinal cohort study of 35 patients with clinically unilateral ES from the private office of one author. Diagnosis of ES was made between 1968 and 1998. On return visits a careful search for ES in the nonexfoliative fellow eye was made by the same author in a semidark room using a slit lamp and maximal pupillary dilatation. Cumulative incidence of converting to bilateral ES was calculated, taking death into account as a competing risk. RESULTS: Cumulative incidence of conversion to bilateral ES was 0.11 (95% CI, 0.03-0.25) at 5 years, 0.36 (95% CI, 0.15-0.57) at 10 years, and 0.52 (95% CI, 0.27-0.93) at 15 years when the day of examination was taken to be the day of conversion, and 0.14 (95% CI, 0.04-0.29) at 5 years and 0.52 (95% CI, 0.26-0.73) at 10 years when analysis was based on conversion taking place at the midpoint between the last two visits. The median age at conversion was 72 years (range, 66 86), and the median age of patients remaining clinically unilateral was 77 years (range, 57-90). Factors associated with time to conversion depended on the strategy of the analysis and included higher age at first visit and larger IOP difference between the involved and uninvolved eye. CONCLUSION: A large proportion of patients with clinically unilateral ES will not convert to bilateral involvement in long-term follow-up. Immunohistochemical studies suggest that local factors must either expedite ES or slow it down in one eye to explain the frequently longstanding asymmetric involvement, and a difference in initial aqueous outflow facility might be examined as one candidate. PMID- 15118460 TI - Corneal thickness in congenital glaucoma. AB - PURPOSE: To compare central corneal thickness between eyes with congenital glaucoma and normal controls and to correlate this parameter with corneal diameter and axial length. METHODS: Eyes of consecutive children with congenital glaucoma with previous glaucoma surgery and eyes of children with inadequacy of lacrimal drainage system with age less than 3 years old were examined under inhalatory general anesthesia. Complete ophthalmologic examination, central corneal thickness, axial length, and corneal diameter measurements were performed. All patients presented with intraocular pressure (IOP) less than 21 mm Hg and no clinical sign of corneal edema. RESULTS: Fifty-five eyes of 55 patients (30 congenital glaucoma and 25 controls) were examined (mean age = 16.6 +/- 10.6 months; 20 female/ 35 males). There was no significant difference in age and gender between glaucoma patients and normal subjects. Mean IOP was higher in glaucomatous eyes (P = 0.02). Corneal diameter and axial length between glaucomatous eyes and controls were significantly different (P < 0.0001 for both). Central corneal thickness was significantly thinner in glaucomatous eyes (P = 0.01). There was a significant correlation between corneal diameter and central corneal thickness and also between central corneal thickness and axial length (r2 = 0.32 and r2 = 0.18, respectively; P < 0.0001 for both). CONCLUSION: Central corneal thickness was significantly thinner in children with congenital glaucoma. This finding may be another confounding factor when measuring IOP in those patients. Pachymetry should be considered during their examination. PMID- 15118461 TI - Association of magnetic resonance imaging of anterior optic pathway with glaucomatous visual field damage and optic disc cupping. AB - PURPOSE: To investigate the association of magnetic resonance imaging (MRI) of anterior optic pathway with glaucomatous visual field damage and optic disc cupping. SUBJECTS AND METHODS: Twenty-three healthy volunteers (controls) and 31 glaucoma patients (14 with primary open angle glaucoma and 17 with normal tension glaucoma) were enrolled. All the participants showed no abnormal signs in their intracranial space and optic tract causing optic nerve atrophy and visual field defect, as confirmed by MRI. Multislice T1-weighted spin-echo imaging was performed in the sagittal plane followed by the coronal plane. MRI enabled the evaluation of the diameter of the optic nerve located in the retro-bulb space and the height of the optic chiasm in an observer-masked fashion. The MRI data were compared with the mean deviation (MD) score of the full threshold static visual field test and the optic cup-disc ratio (C/D ratio). RESULTS: The optic nerve diameter was significantly smaller in glaucoma patients (2.25 +/- 0.33 mm) than in controls (2.47 +/- 0.24 mm) and the height of the optic chiasm was significantly shorter in glaucoma patients (2.12 +/- 0.37 mm) than in controls (2.77 +/- 0.36 mm). The optic nerve diameter showed significant correlation with MD score (r = 0.547, P = 0.001) and C/D ratio (r = 0.407, P = 0.009). These correlations are similar to that between MD score and C/D ratio (r = 0.490, P = 0.001). The height of the optic chiasm showed significant correlation with MD score (r = 0.503, P = 0.01) and low correlation with C/D ratio (r = 0.339, P = 0.113). CONCLUSION: Glaucoma affects the anterior visual pathway anterogradely at least up to the optic chiasm, and these morphologic changes in the anterior visual pathway are correlated with glaucomatous optic nerve damage. MRI of the anterior visual pathway may be a good tool for evaluating glaucomatous damage objectively. PMID- 15118462 TI - Inhibitory effects of anti-glaucoma drugs on corneal epithelial migration in a rabbit organ culture system. AB - PURPOSE: The possible effects of the anti-glaucoma drugs latanoprost, timolol maleate, and nipradilol on corneal epithelial wound healing were investigated in a rabbit organ culture system. METHODS: Corneal blocks (approximately 2 x 4 mm) isolated from albino Japanese rabbits were cultured for 24 hours in medium containing various concentrations (0.01 to 10 mM) of latanoprost, timolol maleate, or nipradilol. The specimens were then fixed and embedded in paraffin, and thin sections were prepared. After the removal of paraffin, the sections were stained with hematoxylin-eosin and observed under a light microscope. Images were recorded digitally with a CCD camera, and the distance over which the epithelium had migrated down the sides of each block was calculated with a computer-assisted morphometric program. RESULTS: Corneal epithelial migration was inhibited by each anti-glaucoma drug in a dose-dependent manner. The 50% inhibitory doses (ID50) for latanoprost, timolol maleate, and nipradilol were 0.362, 0.585, and 4.89 mM, respectively. Relative to the concentrations present in commercially available eyedrops, ID50 corresponded to concentration ratios for timolol, nipradilol, and latanoprost were of 0.037, 0.64 and 3.1, respectively. CONCLUSIONS: The anti glaucoma drugs examined inhibited corneal epithelial migration in a rabbit organ culture system. However, it is unlikely that the corneal surface would be exposed to these agents at inhibitory concentrations for a prolonged period in the normal clinical setting. PMID- 15118463 TI - Antiglaucoma eye drop pulses--increased interleukin-6 secretion by Tenon's capsule fibroblast cultures. AB - PURPOSE: Long-term antiglaucoma eye drop therapy prior to trabeculectomy is a risk factor for surgical failure resulting from Tenon's capsule fibrosis at the fistula site. The study tested the hypothesis that secretion of the proinflammatory cytokine interleukin (IL)-6 by wounded Tenon's capsule fibroblasts is elevated by prior long-term antiglaucoma eye drop treatment. METHODS: Fibroblast cultures were established from Tenon's capsule biopsies during trabeculectomy. Twice daily and for four and a half days, confluent secondary (2-4 passages) cultures were treated (30 minutes at 37 degrees C) with the following drugs (diluted at 1:400-1:100): 0.2% brimonidine-tartrate (Alphagan), 2.0% dorzolamide-HCl (Trusopt), 0.5% timolol-maleate (Timoptic), 2.0% dorzolamide-HCl/0.5% timolol-maleate (Cosopt), 2 and 4% pilocarpine-HCl (Akarpine and Pilocar), 0.005% latanoprost (Xalatan), placebos for Trusopt and Timoptic, and 0.01% benzalkonium chloride. Subsequently, cultures were wounded by removing cells grown on half of each culture dishes along with the medium and conditioned for 20 hours in serum-free growth medium, which was then collected for ELISA for IL-6 (and TNF-alpha and IL-1 beta). Cultures were grown for four additional days to show the maintenance of culture sterility. RESULTS: Latanoprost, pilocarpine HCl, and timolol-maleate increased IL-6 levels in the conditioned medium in a dilution factor-dependent manner (P < 0.05, ANOVA). IL-6 concentrations were increased most significantly by latanoprost and were (pg/ml; mean +/- SEM; N = 3 cultures) 186 +/- 37, 187 +/- 33, 295 +/- 46 and 336 +/- 76 in cultures treated at 1:400, 1:250, 1:150, and 1:100 dilutions, respectively, whereas those of six control cultures averaged 80 +/- 9. Benzalkonium chloride, brimonidine-tartrate, dorzolamide-HCl, Cosopt placebo, Timoptic placebo, and dorzolamide-HCl/timolol maleate did not significantly elevate IL-6 concentrations. IL-1 beta and TNF alpha were not detected in the medium of control cultures and those treated with pilocarpine (1:200). CONCLUSIONS: The present study demonstrated for the first time that the level of IL-6 secretion by wounded Tenon's capsule fibroblast cell cultures was increased by repeat pulsing of these cultures with some, but not all, antiglaucoma eye drops prior to wounding. PMID- 15118464 TI - Effects on aqueous flow of dorzolamide combined with either timolol or acetazolamide. AB - PURPOSE: To determine the effect on aqueous flow of topical dorzolamide 2%, topical timolol 0.5%, or oral acetazolamide 250 mg when used alone or when dorzolamide is combined with either timolol or acetazolamide. METHODS: In 30 patients with ocular hypertension, aqueous flow and intraocular pressure (IOP) were determined at baseline and on the following combinations of drugs in a crossover design: (1) vehicle alone, (2) dorzolamide alone, (3) acetazolamide alone, (4) timolol alone, (5) dorzolamide + acetazolamide, and (6) dorzolamide + timolol. Treated eyes were compared with control eyes and comparisons were made between treatments. RESULTS: Compared with baseline, significant (P < 0.04) IOP reductions in the order of efficacy were: dorzolamide + timolol > dorzolamide + acetazolamide = acetazolamide = timolol > dorzolamide. Aqueous flow was reduced more by dorzolamide + timolol than by each drug alone (P < 0.04) and more by dorzolamide + acetazolamide than by dorzolamide alone (P < 0.04). CONCLUSION: The combination of dorzolamide and timolol demonstrated significant aqueous flow additivity and had greater IOP efficacy than the combination of dorzolamide and acetazolamide. PMID- 15118465 TI - Postoperative pressure regulation in glaucoma shunt surgery: focal tube constriction is not the answer. AB - PURPOSE: To evaluate, in a laboratory setting, the accuracy and the clinical use of focal tube constriction as a means to regulate intraocular pressure after glaucoma drainage implant surgery. MATERIALS AND METHODS: A silicone tube identical to the one used in the Baerveldt and other glaucoma drainage implants was connected to a syringe-pump delivering a continuous flow of demineralized water at a rate of 2.5 microliters per minute. Focal constriction of the tube was obtained by a ring made of a shape-memory metal alloy designed for this purpose. After complete occlusion of the tube lumen by crimping the ring with calibrated pliers, the ring was opened in a stepwise manner by heating it with an argon laser beam. In a second experiment the tube lumen was constricted by placing the tube between the jaws of a micrometer. RESULTS: In both experiments a stepwise lowering of the pressure could be obtained. The resulting pressure levels, however, lacked consistency and predictability to such an extent that clinical application is not feasible. CONCLUSION: It is not possible to regulate pressure in a reliable and predictable way merely by constricting the tube lumen of glaucoma drainage implants. PMID- 15118466 TI - Trends in glaucoma surgery before and after the introduction of new topical glaucoma pharmacotherapies. AB - OBJECTIVE: Recent advances in pharmacotherapy that have improved the ability to effect sustained reductions in intraocular pressure may delay or obviate the need for surgery for open-angle glaucoma. This analysis explored this possibility by evaluating the frequency of surgeries for open-angle glaucoma in the US Medicare population before and after the 1996 introduction of latanoprost, a topical prostaglandin analogue with potent ocular antihypertensive efficacy and a better safety profile than older topical glaucoma medications. METHODS: Data from the Medicare 5% Standard Analytical File were used to develop national estimates of the frequency of glaucoma surgeries in the US yearly from 1994 through 1999. RESULTS: While the number of US Medicare patients with a diagnosis of glaucoma remained constant from 1994 through 1999, the number of inpatient and outpatient glaucoma surgeries declined with a particularly sharp drop between 1996 and 1997. In 1999 relative to 1994, the number of annual glaucoma surgeries among unique patients with a diagnosis of glaucoma was reduced by 72% for inpatient procedures and 42% for outpatient procedures. CONCLUSIONS: The number of glaucoma surgeries among US Medicare patients markedly decreased from 1994 to 1999, during which time the prevalence of glaucoma remained stable in this population. The decrease in surgeries coincided with the introduction of improved topical pharmacotherapies for the management of glaucoma. Although the ability to infer a causal relationship between introduction of new pharmacotherapy and the decrease in surgeries is limited, the consistency of these data with those of several other studies renders the findings compelling. PMID- 15118467 TI - Use of Tisseel fibrin sealant to manage bleb leaks and hypotony: case series. PMID- 15118468 TI - Comparison between results of trabeculectomy in primary congenital glaucoma with and without the use of mitomycin C. AB - PURPOSE: To evaluate the results of trabeculectomy with and without the use of mitomycin C in children with primary congenital glaucoma. METHODS: The authors retrospectively studied the data of 91 patients who underwent the first trabeculectomy for primary congenital glaucoma, with 19 years of follow-up. Success criteria were set with two distinct values of intraocular pressure: lower or equal to 15 mm Hg or lower than 21 mm Hg and separately analyzed. RESULTS: Among the studied patients, 61 had undergone trabeculectomy without mitomycin C and 30 with mitomycin C. The comparison between the groups of patients showed age homogeneity (P = 0.152) and did not demonstrate any difference in preoperative (P = 0.234) and postoperative (P = 0.907) intraocular pressure. Success rates through time, for both pressure limits was not different between the groups. Both age and the presence of previous trabeculotomy did not influence the success of trabeculectomy through time, for the two pressure limits considered. The complication rate was higher among the patients who received mitomycin C (P = 0.010). CONCLUSIONS: The success of trabeculectomy for primary congenital glaucoma with mitomycin C was not different than that of trabeculectomy with mitomycin C in the studied patients. Mitomycin C was associated with a higher incidence of complications. PMID- 15118469 TI - A prospective, comparative study between endoscopic cyclophotocoagulation and the Ahmed drainage implant in refractory glaucoma. AB - PURPOSE: To compare endoscopic cyclophotocoagulation (ECP) and the Ahmed drainage implant in the treatment of refractory glaucoma. METHODS: Sixty-eight eyes of 68 patients with refractory glaucoma were prospectively assigned to either ECP or Ahmed tube shunt implantation. All procedures were performed by a single surgeon. Eyes that were included were pseudophakic with a history of at least one trabeculectomy with antimetabolite, an intraocular pressure (IOP) equal to or above 35 mm Hg on maximum tolerated medical therapy, and a visual acuity better than light perception. Exclusion criteria included eyes that had had previous glaucoma drainage device implantation or a cyclodestructive procedure. Success was defined as an IOP more than 6 mm Hg and less than 21 mm Hg, with or without topical anti-hypertensive therapy. RESULTS: The mean follow-up was 19.82 +/- 8.35 months and 21.29 +/- 6.42 months, for the Ahmed and ECP groups, respectively (P = 0.4). The preoperative IOP, 41.32 +/- 3.03 mm Hg (Ahmed) and 41.61 +/- 3.42 mm Hg (ECP) (P = 0.5), and the mean postoperative IOP, at 24 months follow-up, 14.73 +/ 6.44 mm Hg (Ahmed) and 14.07 +/- 7.21 mm Hg (ECP) (P = 0.7), were significantly different from baseline in both groups (P < 0.001). Kaplan-Meier survival curve analysis showed a probability of success at 24 months of 70.59% and 73.53% for the Ahmed and ECP groups, respectively (P = 0.7). Complications included choroidal detachment (Ahmed 17.64%, ECP 2.94%), shallow anterior chamber (Ahmed 17.64%, ECP 0.0%), and hyphema (Ahmed 14.7%, ECP 17.64%). CONCLUSION: There was no difference in the success rate between the Ahmed Glaucoma Valve and ECP in refractory glaucoma. The eyes that underwent Ahmed tube shunt implantation had more complications than those treated with ECP. PMID- 15118470 TI - Macular and retinal nerve fiber layer thickness measurement reproducibility using optical coherence tomography (OCT-3). AB - PURPOSE: To assess the reproducibility of retinal nerve fiber layer thickness and macular thickness measurements using OCT-3. METHODS: Randomly chosen eyes of healthy individuals were scanned following pupillary dilation by two trained operators (RGO, RV) using OCT-3 (software version A1.1, Carl Zeiss Meditec, Inc., Dublin, CA), three times on separate days within a one-month period. Fast and regular macula (128 A-scans), and fast and regular RNFL (256 A-scans) scanning protocols were performed. Intra- and interoperator measurement reproducibility was evaluated. RESULTS: Ten eyes of 10 subjects (6 females, 4 males) were enrolled. Mean age was 32 +/- 11.2 years (range, 21 to 52 years). Intraoperator reproducibility was high for both macular and RNFL thickness measurements. Mean coefficients of variation (CV) for mean total RNFL thickness measurements ranged from 6.9 +/- 6.4% to 8.0 +/- 3.5% for operators 1 and 2 in fast and regular RNFL protocols. Mean CV for mean macular thickness measurements ranged from 4.7 +/- 2.6% to 6.4 +/- 5.5% for operator 1 and 2 in fast and regular macula protocols. There was no difference in mean total RNFL and mean foveal thickness measurements performed on different days (P > 0.05 for all measurements in all protocols, for operators 1 and 2, ANOVA). Interoperator reproducibility was high for both macular and RNFL thickness measurements (P > 0.05 for all measurements in all protocols, paired t test). CONCLUSION: OCT-3 RNFL and macular thickness measurements are reproducible in normal eyes. These results should be validated in ocular hypertensive and glaucomatous eyes. PMID- 15118471 TI - Validity of rim area measurements by different reference planes. AB - PURPOSE: Reference plane description of the neuroretinal rim in scanning laser tomography should correctly represent optic nerve morphology. We evaluated how well rim area analysis by different reference planes agreed with the appearance of rim area in disc images. METHODS: Three expert observers subjectively and repeatedly analyzed rim area in Heidelberg Retina Tomograph (HRT) images so that each optic disc was measured six times in 100 eyes, 50 normal and 50 glaucoma. Rim area was evaluated globally and in 30 degree sectors. Agreement between rim appearance, as subjectively analyzed, and objective analysis by an experimental reference plane, the standard reference plane, and a reference plane fixed 320 microm below the reference ring was assessed in HRT images. RESULTS: Subjective analysis of rim area in HRT images was consistent between expert observers. Their analysis of rim appearance agreed more closely with experimental reference plane analysis than analysis by the standard or 320-microm reference planes; this was true globally and in every region of the nerve (P = 0.000). The experimental reference plane yielded higher estimates of rim area than did the standard or 320 microm reference planes. CONCLUSION: There was closer correspondence between the appearance of the neuroretinal rim in images and description by the experimental reference plane compared with description by the standard and 320-microm reference planes. PMID- 15118472 TI - Encapsulated blebs following primary standard trabeculectomy: course and treatment. AB - PURPOSE: To investigate the incidence and course of encapsulated filtering blebs (EB) following primary standard trabeculectomy in eyes with non-complicated glaucoma. MATERIALS AND METHODS: Records of patients who were being followed-up in Ankara Numune Hospital, 1st Eye Clinic Glaucoma Department were analyzed and 183 eyes of 183 consecutive patients who underwent primary trabeculectomy without antimetabolites were enrolled in the study. RESULTS: Encapsulation developed in 14 (7.6%) of 183 eyes and identification of bleb encapsulation occurred at a mean follow-up time of 5.1 +/- 5.0 weeks (median = 4 weeks). Mean intraocular pressure (IOP) levels before, during, and after the encapsulated bleb phase were 14.1 +/- 3.6, 23.4 +/- 6.9, and 14.4 +/- 3.9 mm Hg respectively. All eyes responded to conservative medical treatment consisting of aqueous suppressants without digital massage, and encapsulation resolved after a mean duration of 8.3 +/- 4.6 weeks (median = 8 weeks). Patient age, gender, glaucoma diagnosis, and preoperative IOP levels were not significantly related with the occurrence of EB formation. There was no significant difference in the use of topical beta-blockers and miotics in patients that developed EB versus those who did not; however, EB developed only in eyes with prior beta-blocker therapy and no encapsulation occurred in the 16 eyes that had not received topical beta-blockers. Mean duration of follow-up was 45.3 +/- 30.7 months (12 to 127 months) in the group without encapsulation and 44.6 +/- 22.7 months (18 to 84 months) in the EB group. Overall success rates of trabeculectomies in eyes with and without EB were not significantly different (100% and 91.7%, respectively). Complete surgical success, however, without antiglaucomatous medications at the last visit was significantly lower in eyes with prior encapsulation (35.7% with EB, 63.9% without EB) (P = 0.014). CONCLUSION: Filtering bleb encapsulation following primary trabeculectomy responds well to conservative medical IOP-lowering therapy in eyes with glaucoma associated with elevated IOP. PMID- 15118473 TI - Bleb revision for hypotony maculopathy after trabeculectomy. AB - PURPOSE: To assess the efficacy of a bleb revision technique with regard to improvement in intraocular pressure (IOP) and visual acuity after hypotony maculopathy and to evaluate the influence of duration of hypotony on visual outcome. PATIENTS AND METHOD: : Retrospective review of a series of patients who underwent bleb revision for hypotony maculopathy (IOP less than 6 mm Hg and loss of two or more lines of central vision) following trabeculectomy. Bleb revision included excision of avascular bleb tissue, dissection posteriorly between conjunctiva and Tenon's capsule, and advancement and suturing of the conjunctiva at the limbus. RESULTS: Fourteen patients were included in the study. The average age was 49.8 years (range 18-85) and eight were female. Before trabeculectomy, the mean IOP was 23.2 +/- 11.8 mm Hg, and the visual acuity was 20/30-2 or better in 13 patients and 20/80 in one patient. The mean IOP before bleb revision was 1.07 +/- 0.73 mm Hg, and 12 patients had lost an average of 4.33 +/- 2.96 lines of visual acuity from baseline, while one had count fingers and another had hand motion vision. The final mean IOP after bleb revision was 11.07 +/- 3.08 mm Hg. Compared with visual acuity just prior to bleb revision, 12 patients regained an average best corrected vision of 3.08 +/- 2.67 lines, with seven returning to the pre-trabeculectomy vision level. The duration of hypotony prior to bleb revision was 1 to 24 months (average 8.36 months) with no correlation (correlation coefficient -0.13) between duration and visual outcome. CONCLUSION: Bleb revision for hypotony maculopathy following trabeculectomy is an effective technique for raising IOP and limiting visual loss, which is not influenced by duration of hypotony within the time frame of the study. PMID- 15118474 TI - Outcome of eyes undergoing trabeculectomy after intravitreal injections of triamcinolone acetonide. PMID- 15118475 TI - In memoriam: Louis Lasagna, MD 1923-2003. PMID- 15118477 TI - Subjective response to antipsychotics of schizophrenia patients treated in routine clinical practice: a naturalistic comparative study. AB - In routine practice, subjective response to antipsychotics is becoming a critical outcome measure among schizophrenia patients. This study sought to compare subjective response to atypical (risperidone and olanzapine) and typical antipsychotic drugs. Using a naturalistic cross-sectional design, we examined subjective response to antipsychotics (satisfaction with medication and subjective tolerability), psychopathology, side effects, emotional distress, and awareness in schizophrenia patients stabilized on atypical (n = 78) and typical (n = 55) drugs. Analysis of variance and multiple regression analysis were applied. We found that atypical drugs were superior to typical antipsychotics in both measures of subjective response, which were positively correlated (r = 0.52, P < 0.001). Poor subjective response was associated with severity of emotional distress, negative, and activation symptoms in the atypical group and with extrapyramidal side effects and positive symptoms in the typical group. Awareness of treatment is a positive factor that accounted for 20% and 34% of variation in the subjective responses to atypical and typical antipsychotic drugs, respectively. Demographic variables, age of onset, illness duration, and adjunctive drugs did not relate significantly to subjective response to antipsychotic drugs. Thus, atypical drugs are characterized by better subjective response compared with typical antipsychotics; their determinants differed considerably. Satisfaction with medication together with subjective tolerability needs to be considered in clinical trials. PMID- 15118478 TI - A study of the safety, efficacy, and tolerability of switching from the standard delayed release preparation of divalproex sodium to the extended release formulation in patients with schizophrenia. AB - OBJECTIVE: To assess the safety, efficacy, and tolerability of switching from a multiple dose preparation of divalproex sodium delayed release (DR) to once-daily dosing with divalproex sodium extended release (ER) in patients with schizophrenia already receiving the standard DR formulation. METHOD: Thirty subjects with schizophrenia were switched from divalproex DR to a 4-week open label treatment trial of the ER formulation. Baseline plasma levels of valproate were obtained 12 hours postdose. Patients were converted from divalproex DR to ER on a 1.0:1.0 mg basis (rounded up to the nearest 500-mg increment) if baseline valproate plasma levels were > or =85 microg/mL; otherwise, the conversion rate was 1.0:1.2 mg rounded up. Measured at baseline and end point were the Brief Psychiatric Rating Scale and the Udvalg for Kliniske Undersogelser Side Effect Rating Scale. End point plasma levels were obtained at both 12 and 24 hours postdose. RESULTS: Patients who switched from divalproex DR to ER had a small (and probably clinically insignificant) improvement noted on the total Brief Psychiatric Rating Scale at end point (mean change +/- SD = -2.3 +/- 5.4; t = 2.2538; df = 28; P = 0.0322) and on the Udvalg for Kliniske Undersogelser (mean change+/- SD = -2.2+/- 4.1; t = -2.7361; df = 26; P = 0.0111). Baseline and end point trough plasma levels were 80.1 +/- 20.4 and 73.1 +/- 24.2 microg/mL, respectively. Patients who converted on a 1.0:1.0 mg basis had lower end point valproate trough plasma levels than at baseline but did not experience deterioration on their psychopathology. For all patients, end point valproate peak and trough plasma levels were statistically significantly different (t = 3.8706; df = 27; P = 0.0006), but these differences were small in magnitude (mean +/- SD = 14.6 +/-19.6 microg/mL). Seven patients experienced spontaneously reported adverse events, but none required early termination from the protocol. CONCLUSIONS: Switching to a once-daily formulation of ER divalproex can be accomplished without a deterioration in psychopathology. The ER formulation of divalproex sodium appears well tolerated. A parallel group design will be necessary to confirm these findings. PMID- 15118479 TI - Treatment outcome in patients with chronic schizophrenia during long-term administration with risperidone. AB - Treatment outcome was evaluated in outpatients with chronic schizophrenia during long-term administration of risperidone in a study reflecting clinical practice. This UK multicenter, noncomparative, open trial was conducted in 79 patients. Risperidone treatment for 52 weeks commenced at 2 mg/d, with the option to titrate to 6 mg/d. The primary efficacy variable was study failure (study discontinuation for relapse, adverse events, insufficient response, withdrawn consent, lost to follow-up, or noncompliance). The most common dosage was 6 mg/d. Of the 79 patients in the intent-to-treat analysis, 38 completed the study (sustained treatment success), 29 were classified as treatment failure, and 12 were "not evaluable." When the intent-to-treat population was reclassified into study success or study failure, there were 40 study successes (38 treatment successes and 2 ineligible to continue) and 39 study failures (29 treatment failures, 3 lost to follow-up, and 7 noncompliant). Among patients considered to be treatment failures, only 10 had relapse and 10 had adverse events, 2 had insufficient response, and 7 withdrew consent. Raw mean time to treatment failure was 101.7 +/- 90.9 days, and median time was 77 days (range 7 to 284). From Kaplan-Meier curves, mean time to treatment failure was 213.3 +/- 12.2 days; the median was longer than the study period. There was significant improvement (reduction in severity of symptoms) for the intent-to-treat population in total and all Positive and Negative Syndrome Scale subscale scores (P < or =0.0119), Clinical Global Impression Severity (P = 0.0003), cognitive function "letter fluency totals" (P = 0.0044), Abnormal Involuntary Movement Scale (P < 0.0001), and Targeting Abnormal Kinetic Effects scale (measures abnormal kinetic effects; P < 0.0001) at study end point. Most patients considered the treatment at least "acceptable" during the study, and mean change at study end corresponded to between "acceptable" and "quite acceptable." After 1 year of risperidone treatment in a naturalistic setting reflecting usual UK clinical practice, patients with chronic schizophrenia showed improvement in symptoms and reduction in disease severity, and only 10 of 79 had relapse; 48.1% of patients were considered to be sustained treatment successes, 36.7% as treatment failures, and 15.2% as not "evaluable." Fifty-one percent of patients were considered to be study successes and 49% as study failures. PMID- 15118480 TI - Lack of pharmacologic interaction between paroxetine and alprazolam at steady state in healthy volunteers. AB - This investigation aimed to provide evidence on the lack of pharmacokinetic interaction of paroxetine (20 mg/d) and alprazolam (1 mg/d) in combined therapy. In addition, the central effects of both drugs when administered alone and in combination were assessed to rule out any relevant synergistic depressant central effect. Twenty-five healthy young adult volunteers participated in a double blind, double-dummy, placebo-controlled, repeated dose (15 days), 4-period crossover study. Each subject received each of 4 treatment sequences (ie, paroxetine-alprazolam placebo, alprazolam-paroxetine placebo, paroxetine alprazolam, and paroxetine placebo-alprazolam placebo) in randomized order. The ratios for area under the curve within a dosing interval and maximum plasma concentration of the paroxetine plus alprazolam sequence to single agent paroxetine were 1.07 (90% confidence interval = 0.99 to 1.16) and 1.05 (90% confidence interval = 0.97 to 1.13), respectively, with no statistically significant differences between the 2 treatments. Similarly, for alprazolam, ratios for the combined to the single treatment sequence were 0.99 (90% confidence interval = 0.93 to 1.05) and 1.00 (90% confidence interval = 0.94 to 1.07) for area under the curve within a dosing interval and maximum plasma concentration, respectively, showing no evidence for interaction. Comparative pharmacodynamics on the combination was assessed using 6 Psychomotor Performance Tests and 5 Visual Analogue Scales focused on mood variables. Alprazolam and paroxetine plus alprazolam induced similar and significant performance impairment and sedation after both single and repeated dose administration, being less evident on day 15. After dosing, paroxetine plus alprazolam showed a lower recovery pattern than alprazolam alone, especially on day 15. No treatment sequence showed cumulative effects after repeated dose administration. Psychomotor Performance Tests and Visual Analogue Scales data suggested lack of pharmacodynamic interactions. Accordingly, study results showed no evidence for pharmacologic interactions between paroxetine and alprazolam at steady state. The most commonly reported adverse event was drowsiness, with a higher incidence under both single and combined alprazolam treatments. PMID- 15118481 TI - Modulation of hyperreactivity to 35% CO2 after one week of treatment with paroxetine and reboxetine: a double-blind, randomized study. AB - The effects of short treatments (7 days) with paroxetine and with reboxetine on the reactivity to inhalations of 35% carbon dioxide (CO2)/65% oxygen (O2) were compared in 28 patients with Panic Disorder who had positive responses to 35% CO2 inhalations. A double-blind, randomized design was applied. Each patient was given the 35% CO2 challenge on days 0 (before starting the treatment) and 7. Anxiety reactivity to CO2 decreased significantly with both drugs but the decrease was significantly stronger in the group treated with paroxetine. The rate of patients whose reactivity has reduced of at least 50% after 7 days was significantly higher in the group treated with paroxetine (10/14, 71.5%) than in the one treated with reboxetine (3/14, 21.5%). These results indicate that the modulation of the serotonergic system is more relevant for CO2 hyperreactivity than the modulation of the noradrenergic one. PMID- 15118482 TI - Stereoselective metabolism of citalopram in plasma and cerebrospinal fluid of depressive patients: relationship with 5-HIAA in CSF and clinical response. AB - Plasma and cerebrospinal fluid (CSF) concentrations of the enantiomers of citalopram (CIT), its N-demethylated metabolite demethylcitalopram (DCIT) and its deaminated metabolite citalopram propionic acid derivative (CIT-PROP) were measured in plasma and CSF in 22 depressed patients after a 4-week treatment with 40 mg/d citalopram, which was preceded by a 1-week washout period. CSF 5 hydroxyindoleacetic acid (5-HIAA) and homovanillic acid (HVA) were measured at baseline and after the 4-week CIT medication period. Patients were assessed clinically, using the Hamilton Depression Rating Scale (21-item HAM-D): at baseline and then at weekly intervals. CSF concentrations of S-CIT and R-CIT were 10.6 +/- 4.3 and 20.9 +/- 6 ng/mL, respectively, and their CSF/plasma ratios were 52% +/- 9% and 48% +/- 6%, respectively. The CIT treatment resulted in a significant decrease (28%) of 5-HIAA (P < 0.0001) and a significant increase (41%) of HVA in the CSF. Multiple linear regression analyses were performed to identify the impact of plasma and CSF CIT enantiomers and its metabolites on CSF monoamine metabolites and clinical response. There were 10 responders as defined by a > or =50% decrease of the HAM-D score (DeltaHAM-D) after the 4-week treatment. DeltaHAM-D correlated (Spearman) significantly with CSF S-CIT (r = - 0.483, P < 0.05), CSF S-CIT-PROP (r = -0.543, P = 0.01) (a metabolite formed from CIT by monoamine oxidase [MAO]) and 5-HIAA decrease (Delta5-HIAA) (r = 0.572, P = 0.01). The demonstrated correlations between pharmacokinetic parameters and the clinical outcome as well as 5-HIAA changes indicate that monitoring of plasma S CIT, CSF S-CIT and CSF S-CIT-PROP may be of clinical relevance. PMID- 15118483 TI - A placebo-controlled study of nefazodone for the treatment of chronic posttraumatic stress disorder: a preliminary study. AB - Nefazodone is a unique serotonergic antidepressant that acts as both a presynaptic serotonin reuptake inhibitor and a postsynaptic 5-hydroxytryptamine 2A receptor antagonist. Based on the positive results of open-label trials of nefazodone, including one from our group, we tested nefazodone's efficacy in the treatment of posttraumatic stress disorder (PTSD) under placebo-controlled conditions. Forty-one patients with chronic PTSD, predominantly male combat veterans, were enrolled in a randomized, double-blind, placebo-controlled 12-week trial of nefazodone. The primary outcome measure was the Clinician-Administered PTSD Scale. Fifteen patients were randomized to placebo and 26 were randomized to nefazodone. In a repeated-measures analysis of variance with last observation carried forward, patients on nefazodone showed a significant improvement in the percentage change of Clinician-Administered PTSD Scale Total score from baseline compared with those on placebo (P = 0.04; effect size = 0.6). Sample size was not powered to test group differences in the Clinician-Administered PTSD Scale criterion B, C, or D subscale. However, the criterion D subscale showed significant improvement in patients treated with nefazodone compared with those treated with placebo (P = 0.007). In addition, the Hamilton Rating Scale for Depression showed significant improvement compared with placebo (P = 0.008). The nefazodone group also reported an improvement on the PTSD Checklist (self-report scale; P = 0.08) and the Clinician-Administered Dissociative States Scale (P = 0.06). This pilot study supports the efficacy of nefazodone for the treatment of PTSD. However, larger placebo-controlled studies in more diverse patient population are warranted. PMID- 15118484 TI - Clinical rating scales and instruments: how do they compare in assessing abnormal, involuntary movements? AB - OBJECTIVE: Recent studies have shown that quantitative instrumental measurements are more sensitive than clinical rating scales to subclinical dyskinesia and parkinsonism. We therefore hypothesized that an instrumental assessment would be more sensitive to the presence of dyskinetic and parkinsonian movements than the Abnormal Involuntary Movement Scale (AIMS), the Dyskinesia Identification Scale, Condensed User Version (DISCUS), and the Simpson-Angus Scale (SAS). We also hypothesized that the DISCUS, by virtue of its more detailed protocol, would be more sensitive than the AIMS. METHOD: Using blinded raters, we compared the clinical rating scales with instrumental measurements in 100 patients referred to a movement disorders clinic. We collected demographic data, risk factors for tardive dyskinesia, current medication use, Axis I and III disorders, and an estimate of cognitive functioning using the Mini-Mental Status Examination. RESULTS: There was no significant difference between the AIM and the DISCUS in the identification of dyskinesia. However, an instrumental assessment revealed a significantly greater prevalence of dyskinesia. The Mini-Mental Status Examination was the most prominent predictor of both instrumental and clinical measurements of parkinsonian and dyskinetic movements. CONCLUSIONS: It appears that even trained raters, utilizing standard rating scales, may underestimate the prevalence of some motor abnormalities. Instrumental ratings may be helpful to both the clinician and investigator, particularly when abnormal movements are not clinically obvious. The relationship between cognitive impairment and motor abnormalities remains an important area for further research. PMID- 15118485 TI - Effect of Delta-9-tetrahydrocannabinol and cannabidiol on nocturnal sleep and early-morning behavior in young adults. AB - The effects of cannabis extracts on nocturnal sleep, early-morning performance, memory, and sleepiness were studied in 8 healthy volunteers (4 males, 4 females; 21 to 34 years). The study was double-blind and placebo-controlled with a 4-way crossover design. The 4 treatments were placebo, 15 mg Delta-9 tetrahydrocannabinol (THC), 5 mg THC combined with 5 mg cannabidiol (CBD), and 15 mg THC combined with 15 mg CBD. These were formulated in 50:50 ethanol to propylene glycol and administered using an oromucosal spray during a 30-minute period from 10 pm. The electroencephalogram was recorded during the sleep period (11 pm to 7 am). Performance, sleep latency, and subjective assessments of sleepiness and mood were measured from 8:30 am (10 hours after drug administration). There were no effects of 15 mg THC on nocturnal sleep. With the concomitant administration of the drugs (5 mg THC and 5 mg CBD to 15 mg THC and 15 mg CBD), there was a decrease in stage 3 sleep, and with the higher dose combination, wakefulness was increased. The next day, with 15 mg THC, memory was impaired, sleep latency was reduced, and the subjects reported increased sleepiness and changes in mood. With the lower dose combination, reaction time was faster on the digit recall task, and with the higher dose combination, subjects reported increased sleepiness and changes in mood. Fifteen milligrams THC would appear to be sedative, while 15 mg CBD appears to have alerting properties as it increased awake activity during sleep and counteracted the residual sedative activity of 15 mg THC. PMID- 15118486 TI - Alzheimer patients treated with an AchE inhibitor show higher IL-4 and lower IL-1 beta levels and expression in peripheral blood mononuclear cells. AB - The study evaluates the expression and production of cytokines in peripheral blood mononuclear cells of patients with Alzheimer disease treated or not treated with acetylcholinesterase inhibitor, which enhances neuronal transmission. Cytokines associated with brain inflammation such as interleukin (IL)-1beta, IL 6, and tumor necrosis factor-alpha have been implicated in the regulation of amyloid peptide protein synthesis. The anti-inflammatory cytokine, IL-4, may suppress the activity of IL-1beta. Patients were assessed for clinical and immunologic features at baseline and after 1 month of treatment with Donepezil, an acetylcholinesterase inhibitor. Peripheral blood mononuclear cells were cultured with and without phytohemagglutinin stimulation. IL-1beta and IL-4 levels were measured by enzyme-linked immunosorbent assay. Reverse transcriptase polymerase chain reaction was used to determine the expression of cytokines in peripheral mononuclear cells. Compared with untreated patients and healthy control subjects, IL-1beta levels and expression decreased in Alzheimer disease patients treated with Donepezil (P < 0.001). In contrast, IL-4 levels and expression were significantly higher in Alzheimer patients treated with the acetylcholinesterase inhibitor. This increment was observed in both unstimulated and phytohemagglutinin-stimulated peripheral blood mononuclear cells. PMID- 15118487 TI - The relationship between polycystic ovary syndrome and antiepileptic drugs: a review of the evidence. AB - Polycystic ovary syndrome (PCOS) is a serious endocrine disorder characterized by ovulatory dysfunction and hyperandrogenism that is thought to have a higher prevalence in women with epilepsy and perhaps bipolar disorder. Various theories have been offered to explain this higher prevalence of PCOS and other reproductive disorders in these patient populations, including the effects of the disease itself and of antiepileptic drugs, especially valproate, which may directly cause PCOS or indirectly lead to the disorder by causing weight gain that triggers insulin resistance, increased testosterone levels, and other reproductive abnormalities. A prospective, longitudinal study with larger cohorts in newly diagnosed women with epilepsy or bipolar disorder is needed to definitively characterize the relationship between antiepileptic drugs and PCOS. Until data from such a study are available, physicians need to be aware that there is a possibility of developing symptoms of PCOS in women of reproductive age who are treated with antiepileptic drugs. Despite this concern, the choice of antiepileptic drug for women with epilepsy or bipolar disorder should be based on the most effective agent for controlling neurologic symptoms. PMID- 15118488 TI - Risperidone in the treatment of acute schizophrenia. AB - INTRODUCTION: Atypical antipsychotics have proven efficacy and tolerability in the treatment of schizophrenia. While a lot is known about maintenance treatment with atypical antipsychotics, less is known about their role in the management of acute psychotic decompensations. To evaluate the efficacy of the atypical antipsychotic risperidone, we conducted an open-label observational study among admissions to a secure unit. METHOD: Treatment with risperidone was offered to acutely psychotic schizophrenic patients with the requirement of a minimum score of > or =4 on 2 items of the PANSS positive symptoms subscale. Subjects were treated with 4 to 8 mg risperidone in divided doses (mean dose 5.7 +/- 1.5 mg/d). Benzodiazepines and anticholinergics were allowed as co-medications. Clinical Global Impression ratings and Positive and Negative Syndrome Scale ratings were obtained weekly for 4 weeks. RESULTS: Forty-eight subjects (25 males, 23 females; mean age 36.9 +/- 13.4 years, range 18 to 68 years) participated in this study. The mean duration of treatment was 13.4 +/- 9.7 days (range 1 to 31 days). Risperidone was well tolerated, and 26 (54%) patients completed the study. Reasons for discontinuation were need of intramuscular antipsychotic medication (8 subjects), switch to a different antipsychotic (11 subjects), side effects (1 subject), and noncompliance (2 subjects). Treatment with risperidone reduced the mean CGI score from 5.9 to 4.8 (P < 0.001). Likewise, the total PANSS score improved from 110.9 to 86.0 (P < 0.001) with similar reductions in all subscales. DISCUSSION: Our results demonstrate that risperidone is an effective and well tolerated medication for the pharmacologic management of acutely psychotic schizophrenic subjects. PMID- 15118489 TI - Bupropion sustained release for the treatment of hypoactive sexual desire disorder in premenopausal women. AB - Premenopausal women meeting operational criteria for idiopathic, acquired, global hypoactive sexual desire disorder were studied in a randomized, double-blind, placebo-controlled, multiple-site escalating dose 112-day trial of bupropion sustained release. Outcome was measured by investigator-rating and self administered questionnaires. All measures indicated greater sexual responsiveness in women receiving bupropion. The Changes in Sexual Functioning Questionnaire indicated that bupropion had significant effects on increasing measures of sexual arousal, orgasm completion, and sexual satisfaction. PMID- 15118490 TI - Quetiapine overdose and severe rhabdomyolysis. PMID- 15118491 TI - Pancreatitis associated with quetiapine use. PMID- 15118492 TI - A comparison of incidence of diabetes mellitus between atypical antipsychotic drugs: a survey for clozapine, risperidone, olanzapine, and quetiapine. PMID- 15118493 TI - QTc prolongation in multiple drug overdose. PMID- 15118494 TI - Improvement of pathologic gambling symptoms after administration of sertraline: a case report. PMID- 15118495 TI - A pilot naturalistic follow-up of extended sertraline treatment for severe premenstrual syndrome. PMID- 15118496 TI - Nefazodone-induced liver failure. PMID- 15118497 TI - Effects of donepezil on motor function in patients with Alzheimer disease. PMID- 15118498 TI - Prediction of the development of a first major depressive episode with a rapid eye movement sleep induction test using the cholinergic agonist RS 86. PMID- 15118499 TI - Osteopenia associated with divalproex sodium. PMID- 15118500 TI - One eye versus two: a value-based approach. PMID- 15118501 TI - Value-based medicine and vitreoretinal diseases. AB - PURPOSE OF REVIEW: The purpose of the review is to examine the role of value based medicine and its impact, or potential impact, on vitreoretinal interventions. RECENT FINDINGS: Value-based medicine integrates evidence-based data from clinical trials with the patient-perceived improvement in quality of life conferred by an intervention. Cost-utility analysis, the healthcare economic instrument used to create a value-based medicine database, is being increasingly used to study the cost-effectiveness of vitreoretinal interventions. SUMMARY: Vitreoretinal interventions are generally cost-effective because of the great value they impart to patients. Laser surgical procedures, such as for diabetic retinopathy, threshold retinopathy of prematurity, and exudative macular degeneration appear to be especially cost-effective as a group. PMID- 15118502 TI - Dollars and sight: the economics of ophthalmology. AB - PURPOSE OF REVIEW: The purpose of this paper is to detail the economics of ophthalmology including trends in utilization, cost, and expenditure. RECENT FINDINGS: Eye care accounts for $23.1 billion, or approximately 1.93% of total health care expenditures annually. In 1991, eye care service costs in those 65 years or older were $5.5 billion, comprising 4.7% of Medicare spending. By 1999, the cost of eye care services in the 65 or older group had increased to $6 billion, but comprised only 2.8% of Medicare spending. SUMMARY: Although it is imperative that national health expenditure be controlled, ophthalmology has shouldered a disproportionate share of the cost cutting in recent years. When we consider the advances in ophthalmology and the contributions of ophthalmology to improved quality of life, the financial burden absorbed by ophthalmology is striking. PMID- 15118503 TI - Age-related macular degeneration and depression: a review of recent research. AB - PURPOSE OF REVIEW: The purpose of this review is to summarize current research findings regarding relationships between depression and age-related macular degeneration (AMD). RECENT FINDINGS: The current literature indicates that the prevalence of depression among patients with AMD is fairly high (approximately 30%) and that depression is a major cause of disability among patients with AMD, even when severity of vision loss is considered. Interventions to alleviate the emotional distress associated with vision loss are also discussed. SUMMARY: Several studies indicate that AMD is a risk factor for depression, and this has serious consequences for the quality of life among patients with AMD. PMID- 15118504 TI - Retinal complications after laser-assisted in situ keratomileusis (LASIK). AB - PURPOSE OF REVIEW: This paper reviews the retinal complications that may occur after laser-assisted in situ keratomileusis (LASIK). RECENT FINDINGS: During the review period (1 year), several nonrandomized retrospective studies and case reports/series were published. One study was performed to determine the efficacy and safety of prophylactic laser photocoagulation for retinal breaks in patients with myopia undergoing LASIK. Retinal breaks were identified and treated in 39 eyes (2%). None of the patients developed a rhegmatogenous retinal detachment (RRD) (except one trauma case). Another group studied retinal disease observed in 9239 consecutive eyes after refractive surgery (including LASIK) and found RRD in 11 eyes (0.36%) and choroidal neovascularization (CNV) in 10 eyes (0.33%). Three reports described a total of 16 patients with a previously placed encircling scleral buckle for a RRD who had LASIK to correct myopia. In all patients, the visual acuity (VA) improved. Another study reported the characteristics and surgical outcomes of RRD in myopic eyes after LASIK (33 eyes of 27 patients; frequency 0.08% [33/38, 823]). They found that 45.8% lost two or more lines of VA after vitreoretinal surgery. Two letters described the characteristics and potential mechanisms of a macular lacquer crack (one with subsequent development of subfoveal CNV) in a myopic patients corrected by LASIK. SUMMARY: Serious complications after LASIK are infrequent. A dilated fundus examination is very important before LASIK and in every patient whose VA after LASIK is not as good as expected to avoid delayed referral to a vitreoretinal specialist if necessary. Only prospective studies can determine whether the procedure exacerbates myopic pathology. PMID- 15118505 TI - Combining phacoemulsification and vitrectomy in patients with proliferative diabetic retinopathy. AB - PURPOSE OF REVIEW: Modern vitrectomy and cataract extraction techniques have reached levels not imagined 20 years ago. Combining pars plana vitrectomy with phacoemulsification and posterior capsulectomy now has the potential to condense three separate procedures: pars plana vitrectomy, phacoemulsification, and YAG capsulotomy into one procedure. Interest in combined surgery is increasing as reports in the literature documenting its safety and efficacy continue to be published. The purpose of this article is to review the recent trends in combined surgery. RECENT FINDINGS: Studies have shown that when comparing sequential surgery to combined surgery, there is no significant difference in visual outcome. Multiple reports confirm the safety of combined surgery in patients with proliferative diabetic retinopathy. SUMMARY: In select patients with diabetes, combining vitrectomy with phacoemulsification and posterior capsulectomy allows patients who often have bilateral vitreoretinal disease to come to a stable postoperative vision in the eye requiring vitreous surgery much earlier than three separate procedures spread out over months or years. PMID- 15118506 TI - Myopic choroidal neovascularization: natural course and treatment. AB - PURPOSE OF REVIEW: Choroidal neovascularization (CNV) is the most common vision threatening complication of high myopia. Myopic CNV has recently attracted a lot of attention, mainly because of a variety of newly developed treatments. To evaluate the efficacy of these new treatments against myopic CNV, we need to know more precisely the natural course of myopic CNV. The results of most previously reported studies regarding the natural course of myopic CNV, however, are somewhat contradictory. In this review, we describe the recently reported long term prognosis of myopic CNV and the effectiveness of newly developed treatments as well as an overview of possible future treatments. RECENT FINDINGS: A recent study evaluating the visual outcome of myopic CNV for at least 10 years after onset in a large series of patients reported that the visual prognosis of myopic CNV without treatment is extremely poor, mainly because of the secondary development of chorioretinal atrophy around the area of regressed CNV. Although conventional treatments against myopic CNV, such as laser photocoagulation or surgical extraction of CNV, have not been effective, newly developed treatments such as foveal translocation or photodynamic therapy have had favorable results on myopic CNV in the short-term. Pharmacologic interventions to treat or prevent CNV are also expected in the future. SUMMARY: Because the natural prognosis of myopic CNV is extremely poor, treatment to prevent further visual loss is necessary. Based on the steady and gradual visual decrease in myopic CNV over the long term, however, long-term results are needed before the true effectiveness of the newly developed active treatments can be determined. PMID- 15118507 TI - Update on the ocular manifestations of systemic arterial hypertension. AB - PURPOSE OF REVIEW: In a society with a steadily increasing population of patients with hypertension, a significant mortality risk factor, it is important that clinicians be cognizant of the changes seen in ophthalmic examination. This paper demonstrates both the acute and chronic stages of the disease. RECENT FINDINGS: This paper first presents the history of classifying different stages of hypertensive retinopathy, a condition associated with systemic arterial hypertension, as defined by Keith-Wagener-Barker. The most recent recommendations published in the Seventh Report of the Joint National Committee (JNC) on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure are also described; this report defines the current status and classification of hypertension in the United States, as well as recommendations for its treatment and management. Finally, disease management strategies and currently accepted treatment options are presented. SUMMARY: Since systemic arterial hypertension is a key modifiable risk factor for various diseases such as myocardial infarction and stroke, it is crucial that we detect early signs of hypertension to promote better health for our patient population. PMID- 15118508 TI - Corticosteroids in posterior segment disease: an update on new delivery systems and new indications. AB - PURPOSE OF REVIEW: Corticosteroids are traditionally used for inflammatory disorders because of their ability to diminish neutrophil transmigration, limit access to sites of inflammation, and decrease cytokine production. More recently, however, investigators have focused on the angiostatic and antipermeability properties of corticosteroids for posterior segment diseases such as age-related macular degeneration (AMD), diabetic retinopathy, and macular edema. Both new angiostatic and traditional corticosteroids are currently undergoing evaluation as new delivery techniques such as intravitreal injection and intraocular sustained-release devices facilitate high local angiostatic and antipermeability concentrations while minimizing extraocular toxicity. The purpose of this review is to discuss recent work concerning both the mechanism and effectiveness of these newer treatments. RECENT FINDINGS: Steroids may exert a beneficial effect in AMD-related choroidal neovascular membranes (CNVM) through inhibition of CNVM promoting macrophages and direct inhibition of angiogenic growth factors. They may also alter extracellular matrix turnover and inhibit matrix metalloproteinases involved in CNVM formation. Intravitreal steroid injections potently inhibit experimental CNVM in primates and rats and have shown promise in some early human pilot trials. In proliferative diabetic retinopathy, steroids may directly inhibit growth factors such as vascular endothelial derived growth factor and inhibit leukocytes that play an important role in early microvascular alterations. Intravitreal steroid injections inhibit experimental preretinal neovascularization in pigs and rats, and rubeosis in some early human studies. In addition, the effect of steroids on vascular permeability has led to their use for macular edema from many causes such as diabetes and venous occlusive disease. SUMMARY: The use of steroids to treat a number of retinal diseases is gaining wide spread acceptance. The apparent short-term success must be balanced by the fact that the long-term safety and efficacy have yet to be determined for any of these approaches. A number of large randomized prospective clinical trials of steroid compounds and new delivery systems are currently under way for AMD, diabetic retinopathy, uveitis, and other retinovascular diseases, and hopefully these studies will provide guidance about the use of these new modalities. PMID- 15118509 TI - A review of treatments for macular degeneration: a synopsis of currently approved treatments and ongoing clinical trials. AB - PURPOSE OF REVIEW: The purpose of this report is to review recent literature and ongoing clinical trials of the treatment of neovascular age-related macular degeneration. RECENT FINDINGS: Advances in the understanding of the pathogenesis of age-related macular degeneration and choroidal neovascularization have resulted in the emergence of pharmacotherapies targeting various aspects of angiogenesis. Preliminary results with the new agents have been encouraging. Moreover, longer follow-up and subgroup analyses of the Treatment of Age-Related Macular Degeneration with Photodynamic Therapy and Verteporfin in Photodynamic Therapy studies have suggested new potential indications for photodynamic therapy. SUMMARY: Currently a patient with wet macular degeneration may have multiple investigational options that were unavailable several years ago. Ongoing clinical trials are aimed at determining the long-term safety and efficacy of these new pharmacologic and combination therapy modalities. PMID- 15118510 TI - Age-related macular degeneration and quality of life: how to interpret a research paper in health-related quality of life. AB - PURPOSE OF REVIEW: To review how to critically appraise a research article pertaining to changes in health-related quality of life (HRQoL) related to interventions for age-related macular degeneration (AMD). RECENT FINDINGS: We searched PubMed using a strategy that combined the text-words, "macular degeneration" and "quality of life" (n = 73; January 17, 2004), while limiting the search to "clinical trials" (n = 6; of which 3 were published within the past year). A randomized clinical trial evaluating the efficacy of self-management as an intervention for AMD has been selected to introduce the reader to the concept of how to critically review a research paper pertaining to HRQoL in AMD. Other pertinent articles used in this review include recent results published from the Age-Related Eye Disease Study and the Submacular Surgery Trial. SUMMARY: The NEI VFQ is a reliable, valid, and responsive tool when applied to patients with AMD. Self-management of patients with AMD has been demonstrated to improve their HRQoL by way of an internally valid randomized clinical trial. In this issue of Current Opinion in Ophthalmology, we confront the issue of how to assess the validity and importance of a research paper pertaining to the issue of quality of life. To introduce this topic, we will present a real world clinical example to better understand how quality of life may aid in medical decision making. PMID- 15118511 TI - Advances in knowledge and treatment: an update on endophthalmitis. AB - PURPOSE OF REVIEW: In an area of increased willingness to perform surgery as surgical techniques are refined, comes the unavoidable complication of endophthalmitis. Because complete elimination of postsurgical endophthalmitis appears unattainable, strategies directed at optimal prevention, diagnosis, and management are constantly pursued. RECENT FINDINGS: Recent reports of the various aspects of endophthalmitis were analyzed from the leading peer-reviewed ophthalmic journals. SUMMARY: The author reviews the recent work concerning the factors that affect visual prognosis in postsurgical endophthalmitis. PMID- 15118512 TI - Disparities between ophthalmologists and their patients in estimating quality of life. AB - PURPOSE OF REVIEW: Numerous types of instruments can measure quality of life in a given state of health. This article reviews studies that have measured quality of life of patients and physicians for various types of disease states by using a generic preference-based method known as the time tradeoff method of utility analysis. Of particular interest are comparisons of quality of life estimates between patients and physicians for common ophthalmologic conditions. RECENT FINDINGS: In nearly all of the studies identified, the quality of life estimates of patients differed significantly from those of physicians. In studies involving macular disease, cataracts, and numerous other types of medical conditions, physicians generally underestimated the impact of the condition on patients' quality of life. Less of a disparity between patients and physicians has been reported for estimates of quality of life with glaucoma. Differences in quality of life estimates may be related to poor physician-patient communication. SUMMARY: It is important to appreciate the differences in preferences of patients and physicians. Because only patients themselves know their values and needs regarding treatments and outcomes, physicians should routinely elicit patients' preferences and allow them to participate in decision making about treatment. At the societal levels, patients' preferences should help guide decisions including how to allocate finite health care resources. An important tool for this purpose is cost-utility analysis, which incorporates data on patient preferences, along with survival and cost data, to compare the values of competing health care interventions. PMID- 15118513 TI - Recent developments in the management of choroidal melanoma. AB - PURPOSE OF REVIEW: To describe recent advances in the management of choroidal melanoma. RECENT FINDINGS: During the past 10 years, there has been a trend in the management of choroidal melanoma from enucleation to conservative nonenucleation treatments such as thermotherapy, plaque radiotherapy, charged particle radiotherapy, and local resection. Recently published information regarding conservative management has revealed that plaque radiotherapy combined with thermotherapy offers 97% tumor control, similar to charged-particle radiotherapy. Additionally, patient survival after treatment of medium-size melanoma is similar when comparing plaque radiotherapy versus enucleation. Risk factors identifying small choroidal melanoma include patient symptoms, tumor thickness more than 2 mm, presence of associated subretinal fluid, presence of orange pigment on the tumor surface, and location of the tumor margin at the optic disc. These factors assist in early detection of choroidal melanoma. SUMMARY: The management of choroidal melanoma continues to improve with earlier detection of tumors and better treatment methods. Local tumor control is excellent with radiotherapeutic methods. Future studies should investigate control of subclinical metastasis. PMID- 15118514 TI - Bibliography. Current world literature. Retinal, vitreous and macular disorders. PMID- 15118516 TI - Protein C pathways: bedside to bench. Proceedings of the Fifth Margaux Conference on Critical Illness. December 9-11, 2003. Atlanta, Georgia, USA. PMID- 15118517 TI - Protein C/activated protein C pathway: overview of clinical trial results in severe sepsis. AB - OBJECTIVE: To review the results from clinical trials of treatments for severe sepsis involving the protein C/activated protein C pathway. DATA SOURCE: Published research and review articles (PubMed, from 1985 to 2003) relating to clinical trials of compounds involving the protein C pathway. DATA EXTRACTION AND SYNTHESIS: Protein C is converted to activated protein C when thrombin complexes with thrombomodulin. Sepsis is associated with rapid depletion of protein C and blunted endogenous protein C activation. Treatment with protein C concentrate is followed by increased activated protein C plasma levels and a dose-dependent decrease in d-dimer levels in children with purpura fulminans. This supplementation is safe. A phase III trial of recombinant human activated protein C (drotrecogin alfa [activated]) in severe sepsis demonstrated a 6.1% absolute reduction in 28-day mortality compared with placebo. The short- and long-term survival rates associated with drotrecogin alfa (activated) were better in patients at high risk of death associated with a better cost/effectiveness ratio. Treatment with drotrecogin alfa (activated) was associated with an increased risk of serious bleeding compared with placebo during the 28-day study period (3.5% vs. 2.0%). CONCLUSIONS: Treatment with protein C concentrate is followed by an improvement of the coagulopathy and is safe in children with purpura fulminans; however, a large trial involving a high dose is required to determine its effect on mortality and morbidity. Treatment with drotrecogin alfa (activated) leads to substantial reduction in mortality and has an acceptable risk/benefit ratio in septic patients at high risk of death. PMID- 15118518 TI - Human models of endotoxemia and recombinant human activated protein C. AB - OBJECTIVES: To review human models of endotoxemia with an emphasis on the inflammatory and coagulopathic effects of lipopolysaccharide to describe the possible mechanisms of clinical benefit of recombinant human activated protein C (drotrecogin alfa [activated]) in severe sepsis. DATA SOURCE: A selected review of published literature in English on human endotoxemia models with special attention to studies using drotrecogin alfa (activated) in concert with endotoxemia. DATA EXTRACTION AND SYNTHESIS: After intravenous administration of purified lipopolysaccharide, subjects report constitutional influenza-like symptoms associated with an increase in temperature. Within 1 hr of lipopolysaccharide infusion, total leukocyte counts decline. Sustained leukocytosis, due to an increase in neutrophils counts, follows. An increase in markers of neutrophil activation (e.g., serum elastase) accompanies this leukocytosis. Production of tumor necrosis factor-alpha, interleukin-6, and interleukin-8 is also increased. In association with an inflammatory response, lipopolysaccharide administration also activates the coagulation system. Lipopolysaccharide infusion generates thrombin and initially activates fibrinolysis but subsequently generates inhibitors of the fibrinolytic system. Serum levels of endogenous inhibitors of coagulation (e.g., antithrombin, protein C, tissue factor pathway inhibitor) are not affected. Data are available for 40 subjects who have participated in placebo-controlled studies of drotrecogin alfa (activated) infusion before an intravenous dose of lipopolysaccharide. In these subjects, drotrecogin alfa (activated) has minimal effects on measured variables, including physiologic variables, markers of inflammation, and measures of sepsis induced coagulopathy. CONCLUSION: Human endotoxemia is, at best, an incomplete model of human sepsis. In two studies using recombinant human activated protein C in the setting of human endotoxemia, there were minimal effects on hemodynamics, inflammation, thrombin generation, fibrinolysis, and markers of cellular activation. Other putative mechanisms of recombinant human activated protein C, such as inhibition of apoptosis and leukocyte recruitment, remain to be studied. PMID- 15118519 TI - Endpoints in sepsis trials: more than just 28-day mortality? AB - To determine whether an intervention, either therapeutic or diagnostic, is effective, it needs to be assessed according to a predefined endpoint (or outcome measure), the choice of which will vary according to the aims of the study in question and the anticipated effects of the intervention being tested. Studies can have one of several functions (which are not always mutually exclusive), including providing evidence of biological efficacy, determining a clinically important benefit, and achieving regulatory approval. In trials of therapeutic efficacy in sepsis, mortality rates are a good endpoint because death is common and mortality rates are an unambiguous measure: patients either survive or they do not. However, the time at which mortality should be recorded is less clear cut, and this single endpoint provides no information regarding the biological activity or disease modification effects of the agent under investigation. In this article, we will briefly discuss some of the potential alternative endpoints that could be used in the assessment of antisepsis agents. PMID- 15118520 TI - Endogenous protein C activation in patients with severe sepsis. AB - OBJECTIVE: To review how endogenous protein C activation might change in disease states and to discuss the implications of these findings in the context of severe sepsis. DATA SOURCE: A review of the published literature in PubMed together with data from abstracts from 2001 to present. DATA EXTRACTION AND SYNTHESIS: Activated protein C (APC) supplementation has been shown to significantly reduce mortality in patients with severe sepsis, presumably by virtue of its ability to down-regulate coagulation, inflammation, and apoptosis. In vivo, endogenous APC is generated in the circulation when protein C is activated by the thrombin thrombomodulin complex. Protein C activation is augmented by the endothelial cell protein C receptor. Thus, thrombomodulin and the endothelial cell protein C receptor are components of the endothelium-based "machinery" required for efficient activation of protein C. In healthy individuals, the amount of APC formed is proportional to thrombin levels. In vitro studies have shown that thrombomodulin and the endothelial cell protein C receptor are down-regulated by inflammatory cytokines, and the levels of these receptors are reduced in the endothelium of skin-biopsy specimens in children with severe meningococcal sepsis. However, endothelial studies of excised blood vessels provide only a partial picture of the APC pathway in vivo. Knowledge of endogenous plasma levels of protein C, thrombin, and APC may be helpful in assessing the functional status of the protein C pathway in the systemic circulation. To date, there are few reports available on endogenous APC levels in patients with severe sepsis, perhaps due to the lack of available assays that permit both rapid and accurate measurements. A unique feature of our study is that we have developed an APC assay that, for the first time, permits rapid and accurate measurements of plasma APC levels. Preliminary studies using this assay suggest that adult patients with severe sepsis vary markedly in their ability to generate APC endogenously. These results are intriguing because they suggest that, depending on individual defects in the protein C pathway, some patients have impaired protein C activation and might require APC therapy, whereas others may benefit from administration of protein C. Although the clinical efficacy of recombinant human APC (drotrecogin alfa [activated]) in severe sepsis has been reported in a phase III clinical trial, the efficacy of protein C in severe sepsis remains to be determined. CONCLUSIONS: Preliminary results suggest that adult patients with severe sepsis vary markedly in their ability to convert endogenous protein C to APC. Additional research is required to establish whether endogenous APC activation profiles are useful in the clinical management of patients with severe sepsis. PMID- 15118521 TI - Why do animal models (sometimes) fail to mimic human sepsis? AB - OBJECTIVE: To describe potential mechanisms that may account for the observation that drugs that work in treating sepsis in animal models often fail in sepsis trials in patients. DATA SOURCE: MEDLINE searches were used to provide the key information. DATA SUMMARY: Most animal studies are performed acutely in young healthy animals, whereas a significant percentage of the clinical population is elderly with many secondary complications (e.g., diabetes, systemic vascular disease, high blood pressure, immune suppression, cancer). Furthermore, unlike the acute challenge presented in most animal studies, many of the septic patients' clinical histories indicate a relatively slow onset of the disease. In many animal studies, intervention occurs before or during the very early stages of sepsis, when inflammatory cytokine levels are still rising and both organ damage and vascular leakage are minimal. In contrast, current treatment strategies are started when many (probably most) patients are switching from a proinflammatory cytokine response to an anti-inflammatory response and organ damage is already apparent. Patients are also generally receiving some form of supportive therapy (e.g., fluids, vasopressors, ventilators). Because these are seldom used in the animal model, their effect on a particular drug response is difficult to assess. In the animal model, a well-defined bacterial strain, endotoxin challenge, or, in the most complex case, cecal ligation puncture at a defined site is employed to bring about the onset of sepsis. Generally, bacterial proliferation can be controlled in these situations by selecting the appropriate antibiotic (if desired). In human sepsis, the pathogenic bacteria are often not known, mixed infections involving both Gram-negative and Gram-positive bacteria are common, and antibiotic treatment is incomplete or ineffective. Anti inflammatory strategies that impair bacterial killing may be helpful in cases in which antibiotics were effective and harmful when they were not. Thus, an intervention in human sepsis is attempted at a later stage and under very different conditions than it is during efficacy testing in animal models. CONCLUSIONS: Differences in the nature of the initiating agent causing sepsis and the lack of co-morbidities in the animal models probably contribute to some of the differences in animal studies and clinical trials in sepsis. PMID- 15118522 TI - Severe protein C deficiency predicts early death in severe sepsis. AB - OBJECTIVE: To explore the relationship between measures of baseline disease severity and survival time in patients with severe sepsis. DATA SOURCE: A retrospective evaluation of the placebo group from a large placebo-controlled phase III clinical trial (PROWESS) comprising a total of 840 patients with severe sepsis from 164 medical centers. DATA EXTRACTION AND SYNTHESIS: Data collected included baseline demographics and disease severity measurements, baseline protein C and interleukin-6 levels, 28-day and in-hospital survival rates, and cause of death to 28 days. The survival curve for the placebo patients can be divided into three segments during which the rate of death seemed to be different: the rapid alpha phase (day 0 to day 5), the beta phase (day 6 through day 15), and the gamma phase (day 16 to day 28). The risk of death during each phase was statistically significantly different. More patients died of refractory shock during the alpha phase than in the beta and gamma phases, whereas more patients died of respiratory failure during the beta and gamma phases than during the alpha phase. Multiple organ failure was a frequent cause of death during all phases. Protein C levels at the start of each time interval were highly predictive of outcome within that phase, with continued protein C deficiency being associated with mortality. Patients who died during either the alpha or beta phases had higher interleukin-6 levels at baseline than those who died later or who eventually survived. CONCLUSIONS: The rate and cause of death for patients with severe sepsis differs during the 28-day postdiagnosis period. Severe protein C deficiency (<40% of the level of protein C in pooled normal human plasma) and high interleukin-6 levels were associated with early death that resulted predominantly from refractory shock and multiple organ dysfunction. PMID- 15118523 TI - Plasma protein C levels in patients with acute lung injury: prognostic significance. AB - Decreased circulating protein C is a marker of a prothrombotic state that has been associated with poor clinical outcomes in sepsis. However, protein C has not been measured in patients with acute lung injury and acute respiratory distress syndrome (ALI/ARDS). In this study, we measured circulating and intra-alveolar concentrations of protein C in 45 patients with ALI/ARDS from septic and nonseptic causes. Plasma protein C levels were lower in ALI/ARDS compared with normal controls. Lower levels of plasma protein C were associated with worse clinical outcomes, including death, fewer ventilator-free days, and more nonpulmonary organ failures, even when only patients without sepsis were analyzed. In summary, the protein C system is markedly disrupted in patients with ALI/ARDS from both septic and nonseptic causes. The protein C system may be a therapeutic target in patients with ALI/ARDS. PMID- 15118524 TI - Factor V Leiden polymorphism modifies sepsis outcome: evidence from animal studies. AB - OBJECTIVE: The high prevalence of the factor V Leiden mutation in certain populations has prompted speculation that the mutation may have been subject to positive selection during evolution, either by providing a survival benefit or by directly enhancing reproductive performance. We investigated the hypothesis that heterozygous factor V Leiden carrier status might protect against the lethal consequences of severe inflammatory disease. DATA SOURCE: Two mouse models (thrombomodulin-deficient TMPro mice and factor V Leiden mice), in which the endogenous protein C anticoagulant pathway is disrupted either at the level of protein C activation (TMPro mice) or at the level of factor V proteolysis by activated protein C (factor V Leiden mice), were employed. The mutant mouse strains were subjected to lethal doses of bacterial lipopolysaccharide. The effects of these two mutations on coagulation activation and inflammatory cytokine elaboration were observed and compared with those in wild-type mice. DATA SUMMARY: As has already been shown, heterozygous factor V Leiden carrier status improves the survival of mice subjected to endotoxemia induced by bacterial lipopolysaccharide. The survival of homozygous factor V Leiden mice did not differ from that of normal mice. The survival benefit derived from heterozygous factor V Leiden carrier status was only evident at doses of lipopolysaccharide producing death in approximately 50% of wild-type animals. At higher (LD90) or lower (LD10) doses of lipopolysaccharide, the survival of heterozygous factor V Leiden mice did not differ from that of wild-type mice. Concomitant administration of an LD90 dose of lipopolysaccharide and therapeutic heparin abolished the relative survival advantage of heterozygous factor V Leiden mice. Analysis of systemic coagulation and cytokine variables failed to provide conclusive evidence for altered coagulation activation or inflammatory cytokine production as the basis for the survival advantage associated with heterozygous factor V Leiden carrier status. CONCLUSIONS: The improved survival of mice heterozygous for the factor V Leiden mutation complements results from the analysis of the factor V Leiden subgroup of patients enrolled in the PROWESS trial. Such convergent findings in two different species strongly suggest that the factor V Leiden mutation is indeed a potent modifier of the response to severe inflammatory disease. The striking magnitude of the factor V Leiden survival benefit in the initial PROWESS population, and in mice, suggests that the as-yet unknown mechanism conferring this benefit is a rather potent endogenous modifier of the pathogenic pathways engaged in sepsis. Delineation of this pathway will be important for understanding the therapeutic mechanisms, or absence thereof, of agents designed to act at the interface of coagulation and inflammation. PMID- 15118525 TI - Effect of factor V Leiden polymorphism in severe sepsis and on treatment with recombinant human activated protein C. AB - OBJECTIVE: Coagulation activation is part of the acute innate host response to infection that, when uncontrolled, may contribute to organ dysfunction and death. Activated protein C limits excessive coagulation activation by inactivating factors Va and VIIIa. The factor V Leiden mutation (R506Q), a prothrombotic gene polymorphism, disrupts the activity of this natural anticoagulant by rendering factor Va partially resistant to inactivation by activated protein C. Previous findings in the mouse factor V Leiden endotoxemia model and in patients with severe sepsis suggest that factor V Leiden constitutes a rare example of a balanced gene polymorphism that may provide a survival advantage for heterozygous carriers with severe sepsis. We sought to confirm that carriers of this prothrombotic factor V Leiden mutation do not have an increased risk of developing severe sepsis and that carriers with severe sepsis derive similar treatment benefit from recombinant human activated protein C (drotrecogin alfa [activated]) as non-factor V Leiden carriers. DESIGN: Prospective collection of factor V Leiden status from two clinical studies of severe sepsis (PROWESS and ENHANCE). SETTING: : A total of 447 clinical sites across 25 countries. PATIENTS: A total of 3894 adult patients with severe sepsis. INTERVENTION: Either 24 microg x kg x hr drotrecogin alfa (activated) (n = 3063) or placebo (n = 800) for 96 hrs or no exposure to the study drug (n = 31). MAIN RESULTS: The effect of the factor V Leiden carrier status in severe sepsis in the PROWESS study has been previously reported. The combined data on factor V Leiden status from 3894 adult patients with severe sepsis from the PROWESS and ENHANCE (a single-arm, open-label study of drotrecogin alfa [activated]) studies are reported here. At study entry, 3.9% of patients (150/3894) presenting with severe sepsis were heterozygous carriers. No homozygous factor V Leiden carriers were identified. The proportion of factor V Leiden carriers in patients with severe sepsis differs slightly from that predicted (allelic frequency of 2.5%) by the Hardy-Weinberg equation for the general white population (p =.05). There was no significant difference in baseline disease severity (Acute Physiology and Chronic Health Evaluation II score or number of organ dysfunctions) between heterozygous carriers and non Leiden carriers. There was no significant difference in serious bleeding or thrombotic event rates with drotrecogin alfa (activated) treatment between heterozygous carriers and non-Leiden carriers. The 28-day mortality rates for heterozygous carriers and non-Leiden carriers with drotrecogin alfa (activated) treatment were 20.3% and 24.9%, respectively (risk ratio, 0.82; 95% confidence interval, 0.57-1.17). CONCLUSIONS: : Compared with non-Leiden carriers, factor V Leiden heterozygous carriers may have a slightly decreased risk of developing severe sepsis from infection, do not seem to have increased mortality in severe sepsis, and derive similar benefit and risk profiles from drotrecogin alfa (activated) treatment. Therefore, factor V Leiden carriers should not be excluded from this new sepsis therapy. PMID- 15118526 TI - Activated protein C and ischemic stroke. AB - OBJECTIVE: To summarize clinical observations, animal model experimentation, and in vitro studies that advance knowledge of the protein C system, including activated protein C (APC), in the setting of ischemic stroke. DATA SOURCE: Narrative review of selected published primary basic and clinical literature from MEDLINE for 2000-2003. DATA SYNTHESIS: Low levels of plasma APC and a poor response to APC in clotting assays may be markers or risk factors for ischemic stroke. Ischemia during routine endarterectomy causes APC generation in the affected region of the human brain. The prospective epidemiologic Atherosclerosis Risk in Communities (ARIC) study reported that plasma protein C may be protective for ischemic stroke. In murine models of focal cerebral ischemia, APC provided remarkable anti-inflammatory and neuroprotective effects in vivo and increased survival at 24 hrs. Recent in vitro and in vivo studies provide remarkable insights into mechanisms of the neuroprotective activities of APC. Independent of its well-known anticoagulant activity, APC acts directly on cells and alters gene expression profiles, inhibits apoptosis, and down-regulates inflammation. These effects require protease-activated receptor-1 and the endothelial protein C receptor. In an in vitro model involving hypoxia-induced apoptosis of human brain endothelial cells, protease-activated receptor-1 and endothelial protein C receptor were required for APC to exert its anti-apoptotic effects. In these cells, APC blunts hypoxia-induced increases in p53 messenger RNA and protein, reduces pro-apoptotic Bax, and increases anti-apoptotic Bcl-2, thereby inhibiting mitochondrial-dependent apoptosis. Murine ischemic stroke model studies have provided in vivo evidence for the physiologic roles of protease-activated receptor-1 and endothelial protein C receptor in the neuroprotective activities of APC. Because the low doses required for recombinant murine APC to provide neuroprotection do not cause observable anticoagulant effects, the in vivo neuroprotective action of APC seems, at least in part, to be independent of its anticoagulant activity and is likely to involve its anti-apoptotic activity. CONCLUSIONS: There is compelling evidence that ischemic stroke is an attractive target for therapy with APC. PMID- 15118527 TI - Novel functions of thrombomodulin in inflammation. AB - The objective of this study was to review the mechanisms by which thrombomodulin (TM) may modulate inflammation. The data were taken from published research performed by other laboratories and our own experimental results. TM is a transmembrane glycoprotein receptor and cofactor for thrombin in the protein C anticoagulant system. Recent studies have revealed that TM has activities, both dependent and independent of either protein C or thrombin, that affect biological systems beyond the coagulation pathway. This review highlights recent insights, provided by in vitro and in vivo analyses, into how the unique structural domains of TM effectively modify coagulation, fibrinolysis, and inflammation in health and disease. A paradigm is presented to describe how these apparently distinct functions are integrated to maintain homeostasis under stress conditions. Finally, we explore the potential diagnostic and therapeutic utility of dissecting out the structure-function correlates of TM. We conclude that TM plays a central role in regulating not only hemostasis but also inflammation, thus providing a close link between these processes. Elucidation of the molecular mechanisms by which TM functions will likely provide novel targets for therapeutic intervention. PMID- 15118528 TI - Protective role of activated protein C in lung and airway remodeling. AB - Recent studies have implicated the protein C pathway in the mechanism of lung and airway remodeling. The effector enzyme of this pathway is activated protein C (APC). Clinical studies have shown that APC generation is decreased in patients with lung injury and airway inflammation and that this decrease is associated with increased collagen deposition in the lung. In line with these findings, low APC activity has been observed in the bronchoalveolar lavage fluid in animal models of lung injury and airway inflammation. Treatment with APC significantly inhibits the development of lung fibrosis in bleomycin-induced lung injury and the development of airway hyperresponsiveness and allergic inflammation in ovalbumin-induced bronchial asthma. APC may protect the lung from fibrosis and airway remodeling by suppressing activation of coagulation, decreasing the secretion of inflammatory cytokines and platelet-derived growth factor, and promoting fibrinolysis. APC inhibits the expression of cytokines by decreasing the nuclear translocation of signal transducer and activator of transcription 6 and the nuclear factor-kappaB family of transcription factors. In view of its multiple functions, APC constitutes a potential therapeutic agent for inflammatory disorders of the lung and airways. PMID- 15118529 TI - Expression of endothelial protein C receptor and thrombomodulin in the intestinal tissue of patients with inflammatory bowel disease. AB - OBJECTIVE: Inflammatory bowel diseases are characterized by disorders of immunity, thrombosis of large vessels, and microthrombosis of mucosal vessels. The expression of endothelial protein C receptor (EPCR) and thrombomodulin-two receptors of the protein C pathway involved in thrombin scavenging and inflammation-was studied in intestinal resection specimens or mucosal biopsies from patients with inflammatory bowel disease and from controls. The soluble forms of the receptors in plasma were measured. DATA SOURCE: This study involved patients from two large university hospitals. After surgery or biopsy, tissue samples were either frozen or fixed in formalin and embedded in paraffin. Sections for immunohistochemistry examination were cut and tested with the specific antibodies to EPCR and thrombomodulin. RNA was extracted from frozen tissue for amplification via reverse-transcriptase polymerase chain reaction. Normal intestinal and diverticulitis tissue was used as a control. Resection samples from 36 patients with ulcerative colitis, 38 with Crohn's disease, 38 with colonic cancer, and 32 with diverticulitis were studied by immunohistochemistry, and frozen sections from the same patients were studied by immunofluorescence. Twelve biopsy specimens of adjacent intestinal areas from six patients with inflammatory bowel disease were included in the study for reverse transcriptase polymerase chain reaction. Soluble receptors were measured in the plasma of 52 inflammatory bowel disease patients and 52 controls. DATA SUMMARY: EPCR and thrombomodulin were expressed on the mucosal endothelium of controls, and the intensity of the signal decreased in inflammatory bowel disease patients. EPCR was expressed by dendritic-like cells in controls, which also stained positive for CD21. The EPCR/CD21 dendritic-like cells were not as commonly observed in sections from ulcerative colitis patients as they were in sections from control patients (12.0 +/- 3.6 cells per high-power field vs. 23.8 +/- 10.4 cells per high-power field, p =.03), and this decrease was less evident in sections from Crohn's disease patients. Levels of messenger RNA for EPCR paralleled protein expression. Soluble thrombomodulin and EPCR levels were both higher in patients than in controls: 41.5 vs. 26.0 ng/mL (p <.0001) and 141 vs. 130 ng/mL (p <.05), respectively. CONCLUSIONS: EPCR expression on dendritic-like cells that bear the key complement receptor CD21 suggests a role for EPCR in innate immunity. The reduced expression of thrombomodulin and EPCR in the mucosal vessels in inflammatory bowel disease impairs protein C activation, favoring microthrombosis. PMID- 15118530 TI - Endothelium as an organ system. AB - The endothelium is a highly dynamic cell layer that is involved in a multitude of physiologic functions, including the control of vasomotor tone, the trafficking of cells and nutrients, the maintenance of blood fluidity, and the growth of new blood vessels. Over the past several decades, advances in basic research of the endothelium have far outstripped those in the clinic. One explanation for this growing bench-to-bedside chasm relates to the inflexible and largely outdated nature of the present-day medical infrastructure. The constraints of medical subspecialization have created a conceptual blind spot, namely, the inability to appreciate the endothelium for what it is: a cell layer that is teeming with life, every bit as active as any other organ in the body. The overall goal of this review is to bring the endothelium "to life" and to argue that future breakthroughs in biomedicine are contingent on acceptance of the endothelium as a bona fide organ system. PMID- 15118531 TI - Leukocyte and endothelial cell interactions in sepsis: relevance of the protein C pathway. AB - OBJECTIVE: To give an overview of leukocyte and endothelial cell interactions in sepsis and to explore the role of the protein C pathway in modulating the innate immune response via its anti-inflammatory properties. DATA SOURCE: Novel in vitro data and a MEDLINE search for the terms "activated protein C," "recombinant human activated protein C," "inflammation," "leukocyte adhesion," and "sepsis" were used, along with clinical trial databases from the PROWESS trial and a phase I human endotoxin trial evaluating recombinant human activated protein C (drotrecogin alfa [activated]). DATA EXTRACTION AND SYNTHESIS: The protein C pathway is positioned at the interface between the endothelium and the leukocyte response of the innate immune system. Activated protein C (APC) possesses profibrinolytic, anti-inflammatory, and anti-apoptotic properties, acting as an endothelial cell and microvascular modulator in opposition to thrombin and the proinflammatory cytokines. Distribution of the receptor for APC, endothelial protein C receptor, was detected on effector cells of the innate immune response. This suggests a further role for the protein C pathway in regulating inflammation. In neutrophils and eosinophils, an endothelial protein C receptor mediated APC response leads to reduced migration in response to cytokine gradients. Endothelial protein C receptor may also suppress the apoptotic response in monocytes and enhance the expression of the adhesion integrin CD11b in granulocytes. The microvascular, anti-inflammatory influence of APC in sepsis is supported by suppression of endothelial cell adhesion molecules and the ability of APC to protect the endothelium from inflammatory insult. CONCLUSIONS: The coordinated effects of the protein C pathway on the endothelium and the leukocyte response of the innate immune system are supported by potential restriction of endothelial protein C receptor expression to cells of the innate immune system and by suppression of adhesion molecule expression on the endothelium by APC. Reduced neutrophil migration in response to cytokines is also mediated by endothelial protein C receptor. Further clinical studies will be needed to define the intrinsic role of the protein C pathway in coordinating the innate immune response in endothelium-based inflammation. PMID- 15118532 TI - Protease-activated receptor signaling in the regulation of inflammation. AB - OBJECTIVE: To discuss recent studies addressing the relationship between protease activated receptor signaling, coagulation, and inflammation. DATA EXTRACTION AND SYNTHESIS: This review article covers relevant original articles published until October 2003 dealing with animal models, clinical trial data, and in vitro experiments. CONCLUSIONS: Although activation of protease-activated receptors has been implicated in the proinflammatory effects of the coagulation cascade, current data provide evidence that protease-activated receptor signaling plays a more complex role in the regulation of inflammation and endothelial homeostasis. Sensitive assays for coagulation activation have provided clear evidence that targeting the coagulation pathway effectively reduces the coagulopathy in sepsis. However, the effect of these anticoagulant agents on sepsis-associated inflammation is less clear. Further insight into this question will require the development or use of additional biomarkers for assessing pharmacologic interference with coagulation-related cell-signaling pathways. PMID- 15118533 TI - Tissue factor, coagulation proteases, and protease-activated receptors in endotoxemia and sepsis. AB - Inhibition of the tissue factor-factor VIIa complex reduces coagulation and inflammation in animal models of endotoxemia and sepsis and in patients with severe sepsis. However, the mechanism by which tissue factor-dependent activation of the coagulation cascade enhances inflammation is not known. We tested the hypothesis that coagulation proteases enhance inflammation during endotoxemia by activating protease-activated receptors (PARs) within the vasculature. We found that genetically modified mice expressing low levels of tissue factor exhibited reduced interleukin-6 expression and increased survival in a mouse model of endotoxemia compared with control mice. In contrast, hirudin inhibition of thrombin or a deficiency in either PAR-1 or PAR-2 did not affect interleukin-6 expression or mortality. However, combining hirudin treatment to inhibit thrombin signaling through PAR-1 and PAR-4 with PAR-2 deficiency reduced lipopolysaccharide-induced interleukin-6 expression and increased survival. Taken together, our results suggest that activation of multiple PARs by coagulation proteases enhances inflammation during endotoxemia. PMID- 15118534 TI - Structure and functions of the endothelial cell protein C receptor. AB - The endothelial cell protein C receptor (EPCR) plays a critical role in augmenting protein C activation by the thrombin-thrombomodulin complex and in modulating the functions of the protein C pathway to aid in preventing organ damage due to various challenges. EPCR exhibits a sequence and three-dimensional homology with the major histocompatibility class 1/CD1 family of proteins. This family of proteins is characterized by having a deep groove that is usually used in antigen presentation. In the case of CD1c and CD1d, this groove is filled with a lipid antigen, usually a glycolipid. Like the CD1 series, EPCR has a lipid in the corresponding groove. In this case, the lipid is usually phosphatidylcholine, but it may be phosphatidylethanolamine. The bound lipid contributes to protein C binding, but its structure suggests a role in maintaining EPCR structure rather than contributing directly to protein C binding. Potential roles for EPCR in hematopoiesis are suggested by the finding that EPCR is located on hematopoietic stem cells at reasonably high concentrations. The structure and the lipid antigen suggest that EPCR may be involved in preventing autoimmunity, which would be consistent with findings in CD1d knockout mice. Complete deletion of EPCR function results in embryonic death, at least in part due to placental thrombosis. In adult animals, the anticoagulant and anti-inflammatory responses to endotoxin increase with increasing EPCR expression. Some of the anti inflammatory activity is likely to be due to EPCR's interactions with the integrin Mac-1 (CD11b/CD18) on leukocytes, an interaction that probably limits tight adhesion of leukocytes to activated endothelium. Thus, available data suggest a potential role of EPCR in hematopoiesis, autoimmunity, and the control of both the coagulation and inflammation responses to infection and trauma. PMID- 15118535 TI - Effect of drotrecogin alfa (activated) on human endothelial cell permeability and Rho kinase signaling. AB - OBJECTIVE: To explore whether the improvement in organ function and the vasoactive effect observed in the clinical studies of drotrecogin alfa (activated) (recombinant human activated protein C, rhAPC) in sepsis are a result of rhAPC's effect on endothelial cell (EC) permeability and modulation of the intracellular cytoskeleton via the Rho kinase signaling pathway. DESIGN: Findings regarding dose and duration of exposure to the drug with sequential addition of rhAPC and mediators (thrombin, histamine, interleukin-1 beta). SETTING: Research laboratory in a pharmaceutical company. SUBJECTS: Cultured primary human EC from different tissues and vascular beds. INTERVENTIONS: A monolayer of EC was incubated with either rhAPC, thrombin, histamine, or interleukin-1 beta alone or with rhAPC in combination with thrombin or interleukin-beta. The effect of rhAPC and mediators on EC permeability was monitored with measurement of electrical resistance. The effect on Rho kinase pathway signaling was monitored by the levels of phosphorylated myosin light chain and blockage with the Rho kinase specific inhibitor, Y27632. MEASUREMENTS AND MAIN RESULTS: Thrombin alone induced an early, concentration-dependent, and transient leakiness of EC. Interleukin-1 beta (0.5 ng/mL) induced an early, irreversible leakiness of EC. rhAPC (0.05-0.2 microg/mL, approximate median therapeutic blood levels) alone had no effect on EC permeability. rhAPC at > or=1 microg/mL induced an early EC leakage. rhAPC (0.19 microg/mL) attenuated the leakage induced by 0.5 ng/mL interleukin-1beta on microvascular EC derived from lung and skin and partially attenuated the leakage induced by 0.25 nM thrombin on human coronary arterial ECs. Levels of phosphorylated myosin light chain increased rapidly in human coronary arterial ECs when stimulated with thrombin or rhAPC (about 100-fold less potent) in a concentration-dependent manner via the Rho kinase signaling pathway. Short (5 mins) preconditioning of human coronary arterial ECs with 0.19 microg/mL rhAPC partially blocked the increase in phosphorylated myosin light chain levels induced by thrombin (0.06-0.2 nM). CONCLUSIONS: At concentrations exceeding physiologic and therapeutic levels, rhAPC increases EC permeability, an effect not seen at lower concentrations. The data suggest that interpretation of published in vitro and in vivo data of rhAPC and EC permeability should take into consideration the concentrations of rhAPC used or achieved. Other preliminary novel observations suggest that studying the effects of rhAPC on EC permeability and intracellular cytoskeletal organization may provide understanding of the effect of rhAPC on EC function. PMID- 15118536 TI - Beyond sepsis: activated protein C and ischemia-reperfusion injury. AB - OBJECTIVE: To review potential clinical situations beyond sepsis in which activated protein C might be an effective treatment. DATA SOURCE: Published articles between 1970 and 2003 on experimental and clinical studies of activation of both coagulation and inflammation in various disease states. DATA SYNTHESIS AND CONCLUSION: The efficacy of activated protein C in sepsis might rely on the fact that it can modulate both coagulation and inflammation. Therefore, administration of activated protein C could be beneficial in disease states that are also characterized by the simultaneous activation of these systems. Ischemia reperfusion injury of various organs may represent such a state. Indeed, the involvement of the protein C system has been demonstrated in various experimental studies of ischemia-reperfusion, including studies in renal ischemia-reperfusion syndromes, coronary atherosclerosis and acute coronary syndromes, and intestinal ischemia and reperfusion. In some of these models, activated protein C administration, or other interventions in the protein C system, was shown to be beneficial. PMID- 15118537 TI - Importance of hemostatic gene polymorphisms for susceptibility to and outcome of severe sepsis. AB - Individuals vary considerably in their susceptibility to infection and in their ability to recover from apparently similar infectious processes. These differences can be partially explained by polymorphisms of the genes encoding proteins involved in mediating and controlling the innate immune response, the inflammatory cascade, coagulation, and fibrinolysis. It is evident from experimental studies that dysregulation of the coagulation system, which is characteristic of the pathophysiology of septic shock (a procoagulant and antifibrinolytic state), contributes to systemic inflammation and death in sepsis. Several genetic variations in proteins that increase coagulation or impair anticoagulation and fibrinolysis have been described. Thus, polymorphisms have been reported in prothrombin, fibrinogen, factor V, tissue factor, endothelial protein C receptor, and plasminogen activator inhibitor-1 genes. Some of them are associated with an increased risk of pulmonary emboli, acute myocardial infarction, stroke, and severe sepsis. Hence, the deletion polymorphism (4G) within the promoter region of the plasminogen activator inhibitor-1 gene leads to impaired fibrinolysis and influences the severity and outcome of meningococcal disease and the susceptibility to severe sepsis and multiple organ failure after trauma. The factor V Leiden mutation is associated with thrombotic events and has been reported to exacerbate purpura fulminans in meningococcal infection. Surprisingly, this genetic variant seems to provide a survival advantage in severe sepsis, underlying the extreme complexity of the interaction between inflammation and coagulation. The study of genetic polymorphisms might provide important insights into the pathogenesis of severe sepsis and could make it possible to identify individuals who are at risk of developing or dying of severe infections. As genetic associations are discovered, medical practice can become more preemptive, using the predictive ability of genetics to anticipate disease and recommend therapy. PMID- 15118538 TI - Thrombin activatable fibrinolysis inhibitor: not just an inhibitor of fibrinolysis. AB - OBJECTIVE: To review the activation of thrombin activatable fibrinolysis inhibitor (TAFI) and activity of activated TAFI (TAFIa) as it relates to the regulation of both fibrinolytic and proinflammatory substances. DATA SOURCE: Published articles and reviews (from PubMed, published between 1962 and 2003) on experimental studies of coagulation, fibrinolysis, and inflammation. DATA SYNTHESIS AND CONCLUSIONS: The principal physiologic role of TAFI is still a matter of debate. Although TAFI activation can result from proteolysis by a number of proteases, the most likely physiologic activators are thrombin (in complex with the cofactor thrombomodulin) and plasmin (in complex with polysaccharide cofactors). The activated enzyme, TAFIa, displays carboxypeptidase B-like activity and probably regulates both fibrinolysis and inflammation in response to injury and infection. At present, there is limited understanding of the role that TAFI plays in the interrelationships between coagulation, fibrinolysis, and inflammation. Although the potential therapeutic value of TAFIa inhibition/TAFI activation awaits further investigation, the data gathered to date suggest that, like activated protein C, TAFIa may play a pivotal role in regulating the crosstalk between coagulation, fibrinolysis, and inflammation. PMID- 15118539 TI - Radiation injury and the protein C pathway. AB - OBJECTIVE: To summarize current knowledge regarding the role of the thrombomodulin (TM)-protein C system in acute and chronic radiation responses in normal tissues. DATA SOURCE: Studies published in the biomedical literature during the past three decades and cited in PubMed and unpublished clinical and laboratory data from our own research program. STUDY SUMMARY: The risk of injury to normal tissues limits the cancer cure rates that can be achieved with radiation therapy. Microvascular injury is a prominent feature of normal tissue radiation injury and plays a critical role in both acute (inflammatory) and chronic (fibrotic) radiation responses. Evidence from our own and other laboratories strongly suggests that dysfunction of the TM-protein C system plays a key role in the pathogenesis of radiation-induced injury to normal tissue. Exposure of normal tissues to ionizing radiation causes a pronounced, sustained deficiency of endothelial TM. This is likely to be due to a combination of initial inactivation of TM by reactive oxygen species, reduced transcription of TM, and release of TM into the circulation. Deficient levels of endothelial TM cause loss of local vascular thrombo-resistance, excessive activation of protease activated receptor-1 by thrombin, and insufficient activation of protein C. These changes are presumed to be critically involved in many aspects of acute radiation toxicity and in sustaining the fibroproliferative processes that lead to chronic radiation-induced organ dysfunction and clinical complications. CONCLUSION: Injury of vascular endothelium may be key to the acute responses of normal tissues to ionizing radiation and to the progressive nature of chronic radiation fibrosis. Restitution of the TM-protein C pathway is an appealing strategy by which to prevent or treat normal tissue toxicity associated with radiation treatment of cancer. PMID- 15118540 TI - Platelet factor 4 modulation of the thrombomodulin-protein C system. AB - OBJECTIVE: To review published studies of the influence of platelet factor 4 (PF4) and other cationic proteins on the generation of activated protein C (APC) by the thrombomodulin-protein C system. DATA SOURCE: Using the PubMed citation index, literature published from 1973 to 2003 regarding cationic proteins, PF4, and the thrombomodulin-protein C system was reviewed. DATA SYNTHESIS: All other cationic proteins studied to date either impair or do not affect APC generation via the thrombomodulin-protein C system; however, the platelet alpha-granule protein PF4 causes a 25-fold increase in the ability of thrombomodulin polypeptides to generate APC and a ten-fold increase in the ability of cultured endothelial cell-associated thrombomodulin to generate APC. The mechanism underlying this phenomenon depends on binding of the cationic PF4 to the anionic, vitamin K- dependent gamma-carboxyglutamic acid domain of protein C. The extent of PF4's stimulation of APC generation is further increased by its interaction with the anionic glycosaminoglycan moiety that is variably expressed through posttranslational, O-linked glycosylation of thrombomodulin. In an in vivo thrombin-infusion model of thrombomodulin activation in cynomolgus monkeys, previous intravenous infusion of pharmacologic amounts of PF4 resulted in circulating APC levels and APC-dependent prolongation of activated partial thromboplastin times that were two- to three-fold greater than those observed in saline-infused control animals. CONCLUSIONS: These findings raise the possibility that PF4 plays a hitherto unsuspected physiologic role in enhancing APC generation in vivo. They also provide a rationale for considering the infusion of PF4 or PF4-related peptides or peptidomimetics as a way of beneficially stimulating "endogenous" APC generation from circulating protein C in pathologic human disease states such as sepsis. PMID- 15118541 TI - Protein S: a conduit between anticoagulation and inflammation. AB - OBJECTIVE: To review the multifaceted roles of the anticoagulant protein S, facilitating a better comprehension of this protein's role in anticoagulation and inflammation pathways and the crosstalk between these pathologic states. DATA SOURCES AND STUDY SELECTION: Original research and review articles published in English pertaining to protein S, sourced from PubMed, during the last 30 yrs. DATA EXTRACTION AND SYNTHESIS: The protein C anticoagulant pathway is an essential mechanism for attenuating thrombin generation by the membrane-bound procoagulant complexes, tenase and prothrombinase. Protein S is a nonenzymatic protein. In the absence of activated protein C, it demonstrates anticoagulant activity; in the presence of activated protein C, it functions as a cofactor for activated protein C-dependent proteolytic inactivation of the coagulation cofactors factor Va and factor VIIIa. However, in plasma, these anticoagulant activities are limited by the concentration of free protein S (approximately 40% of the total protein S plasma concentration). The remaining protein S (approximately 60%) is found in a high-affinity, calcium-stabilized complex with C4b-binding protein, which renders this fraction devoid of anticoagulant function. Several recent investigations have attributed novel activated protein C independent functions of protein S to the association of protein S with C4b binding protein, thus establishing the importance of this fraction of plasma protein S. CONCLUSIONS: Together, these data support a role for protein S in both anticoagulation and inflammation, facilitating a better understanding of the need for both free and C4b-binding protein-bound protein S. Although these physiologic roles are truly dichotomous in terms of functional end point, mechanistically, both involve high-affinity membrane binding to phosphatidylserine-bearing surfaces. This binding is mediated by the n-terminal gamma-carboxyglutamic acid rich domain of this protein. PMID- 15118542 TI - Sternoclavicular septic arthritis: review of 180 cases. AB - We review 170 previously reported cases of sternoclavicular septic arthritis, and report 10 new cases. The mean age of patients was 45 years; 73% were male. Patients presented with chest pain (78%) and shoulder pain (24%) after a median duration of symptoms of 14 days. Only 65% were febrile. Bacteremia was present in 62%. Common risk factors included intravenous drug use (21%), distant site of infection (15%), diabetes mellitus (13%), trauma (12%), and infected central venous line (9%). No risk factor was found in 23%. Serious complications such as osteomyelitis (55%), chest wall abscess or phlegmon (25%), and mediastinitis (13%) were common. Staphylococcus aureus was responsible for 49% of cases, and is now the major cause of sternoclavicular septic arthritis in intravenous drug users. Pseudomonas aeruginosa infection in injection drug users declined dramatically with the end of an epidemic of pentazocine abuse in the 1980s. Sternoclavicular septic arthritis accounts for 1% of septic arthritis in the general population, but 17% in intravenous drug users, for unclear reasons. Bacteria may enter the sternoclavicular joint from the adjacent valves of the subclavian vein after injection of contaminated drugs into the upper extremity, or the joint may become infected after attempted drug injection between the heads of the sternocleidomastoid muscle. Computed tomography or magnetic resonance imaging should be obtained routinely to assess for the presence of chest wall phlegmon, retrosternal abscess, or mediastinitis. If present, en-bloc resection of the sternoclavicular joint is indicated, possibly with ipsilateral pectoralis major muscle flap. Empiric antibiotic therapy may need to cover methicillin resistant Staphylococcus aureus (MRSA). PMID- 15118543 TI - Coccidioidomycosis: a descriptive survey of a reemerging disease. Clinical characteristics and current controversies. AB - Coccidioidomycosis is a fungal disease with protean manifestations endemic to the Lower Sonoran Life Zone, which includes the hot deserts of the southwestern United States and areas of Mexico. Two hundred and twenty-three patients were found to have coccidioidomycosis at our institution from 1994-2002, the largest reported cohort of coccidioidomycosis patients since the 1950s. Of these patients, 58% presented with isolated pulmonary disease, 14% had high (>1:16) complement fixation titers without clear evidence of dissemination, 22% had definite disseminated disease, and 5% had unclassified disease. Enzyme immunoassay was a reliable diagnostic tool in those with symptomatic disease, but had a low specificity in those who were asymptomatic. Complement fixation titers of > or =1:16 were associated with dissemination to bone or skin but were not helpful in evaluating central nervous system disease. Thirteen percent of patients with high complement fixation titers (>1:16) without clear evidence of dissemination on presentation and 7% of those with isolated pulmonary disease eventually progressed to disseminated disease; 30% of Filipino patients with pulmonary disease progressed to disseminated disease. Nonwhite race was a predictor for dissemination; African American patients more often developed disseminated bony disease while Filipinos were more likely to develop cutaneous or central nervous system disease. Relapse of disseminated coccidioidomycosis occurred in 24% of patients; the risk was highest (71%) among those with central nervous system disease. Azole therapy was generally inferior to amphotericin B in disseminated disease. Predictors of permanent disability included African American or Filipino race, central nervous system disease, and bony disease. PMID- 15118544 TI - Habitual betel quid chewing and risk for hepatocellular carcinoma complicating cirrhosis. AB - This case-control study aimed to assess the independent and interactive role of habitual betel quid chewing and known risk factors for hepatocellular carcinoma (HCC). Subjects enrolled included 210 pairs of sex- and age-matched cirrhotic patients with HCC, patients with cirrhosis alone, and healthy controls. Information on risk factors was obtained through serologic examination of hepatitis B surface antigen (HBsAg) and antibodies to hepatitis C virus (anti HCV), and a standardized personal interview with a structured questionnaire. Multivariate analysis indicated that betel quid chewing (odds ratio [OR], 5.81; 95% confidence interval [CI], 2.26-14.94); HBsAg (OR, 37.98; 95% CI, 19.65 73.42); and anti-HCV (OR, 47.23; 95% CI, 18.86-118.25) were independent risk factors for HCC when HCC patients were compared with healthy controls. Using patients with cirrhosis alone as a reference group, multivariate analysis indicated that only betel quid chewing (OR, 1.69; 95% CI, 1.04-2.76) and HBsAg (OR, 1.54; 95% CI, l.01-2.37) were independent risk factors for HCC. There was an additive interaction between betel quid chewing and the presence of either HBsAg (synergy index, 5.22) or anti-HCV (synergy index, 1.35). Moreover, a higher risk of HCC was associated with a longer duration of betel quid chewing and a larger amount of betel quid consumed (each p(for trend) < 0.0001). In conclusion, betel quid chewing is an independent risk factor for cirrhotic HCC. There is an additive interaction between betel quid chewing and chronic hepatitis B and/or hepatitis C virus infection. PMID- 15118545 TI - Cryptococcus neoformans Infection in Patients With Cirrhosis, Including Liver Transplant Candidates. AB - We reviewed the cases of patients with cirrhosis, including liver transplant candidates, at our institution in the last 3 years (n = 5) and those individually described in the literature (n = 28), to assess unique characteristics and outcome of cryptococcosis in these patients. Sixty-four percent (21/33) of the patients had no other recognized immunosuppression. Peritonitis (in 45%, 15/33 of the patients) with modest pleocytosis in the ascitic fluid, was the most common presenting feature. Median time to detection of Cryptococcus in the ascitic fluid cultures was 6 days. Overall mortality rate was 81% (26/32); death was deemed attributable to cryptococcosis in 24/26 patients who died. Evaluation of culture negative neutrocytic ascites in febrile cirrhotic patients warrants consideration of cryptococcal peritonitis. PMID- 15118546 TI - Clinical course and flow cytometric analysis of paroxysmal nocturnal hemoglobinuria in the United States and Japan. AB - : To determine and directly compare the clinical course of white and Asian patients with paroxysmal nocturnal hemoglobinuria (PNH), data were collected for epidemiologic analysis on 176 patients from Duke University and 209 patients from Japan. White patients were younger with significantly more classical symptoms of PNH including thrombosis, hemoglobinuria, and infection, while Asian patients were older with more marrow aplasia. The mean fraction of CD59-negative polymorphonuclear cells (PMN) at initial analysis was higher among Duke patients than Japanese patients. In both cohorts, however, a larger PNH clone was associated with classical PNH symptoms, while a smaller PNH clone was associated with marrow aplasia. Thrombosis was significantly more prevalent in white patients than Asian patients, and was associated with a significantly higher proportion of CD59-negative PMN. For individual patients, CD59-negative populations varied considerably over time, but a decreasing PNH clone portended hematopoietic failure. Survival analysis revealed a similar death rate in each group, although causes of death were different and significantly more Duke patients died from thrombosis. Japanese patients had a longer mean survival time (32.1 yr vs. 19.4 yr), although Kaplan-Meier survival curves were not significantly different. Poor survival in both groups was associated with age over 50 years, severe leukopenia/neutropenia at diagnosis, and severe infection as a complication; additionally, thrombosis at diagnosis or follow-up for Duke patients and renal failure for Japanese patients were poor prognostic factors. These data identify important differences between white and Asian patients with PNH. Identification of prognostic factors will help the design of prospective clinical trials for PNH. PMID- 15118547 TI - [Neuropsychology of Dementia. Proceedings and abstracts from the Spring Meeting of the French Language Neuropsychology Society. Caen, France, 23-24 May 2003]. PMID- 15118549 TI - [Episodic memory: from mind to brain]. AB - Episodic memory is a neurocognitive (brain/mind) system, uniquely different from other memory systems, that enables human beings to remember past experiences. The notion of episodic memory was first proposed some 30 Years ago. At that time it was defined in terms of materials and tasks. It was subsequently refined and elaborated in terms of ideas such as self, subjective time, and autonoetic consciousness. This chapter provides a brief history of the concept of episodic memory, describes how it has changed (indeed greatly changed) since its inception, considers criticisms of it, and then discusses supporting evidence provided by (a) neuropsychological studies of patterns of memory impairment caused by brain damage, and (b) functional neuroimaging studies of patterns of brain activity of normal subjects engaged in various memory tasks. I also suggest that episodic memory is a true, even if as yet generally unappreciated, marvel of nature. PMID- 15118550 TI - [Dysexecutive syndromes and degenerative diseases]. AB - A dysexecutive syndrome is observed not only in frontotemporal lobar degeneration, but also in subcortical degenerative diseases, and even in Alzheimer's disease whose lesions predominate in temporoparietal associative areas. The association between a dysexecutive syndrome and various cerebral localisations may be explained by the fact that cognitive and behavioral organisation recruits anatomofunctional frontostriatal and frontoparietal circuits. Both animal experimentation and human clinical observation argue in favour of a functional continuity and complementarity among these loops. The prefrontal cortex would be particularly needed in new situations, to inhibit old programs of action not adapted to the present context and to elaborate new ones; the basal ganglia would be rather required by the repetition of the situation to progressively transform the new program in routine. If we refer to Shallice model, we can hypothesize that optimal executive functions require the preservation not only of the Supervisory Attentional System, mainly dependent on the prefrontal cortex, but also of the Contention Scheduling, recruiting the basal ganglia, and of the Schemas of Action, represented in parietal and premotor areas. Therefore, the neuropsychological assessment of patients with degenerative diseases contributes to the understanding of the anatomofunctional architecture of executive functions. PMID- 15118551 TI - [Hallucinations and dementia. Prevalence, clinical presentation and pathophysiology]. AB - Hallucinations are a common feature of certain degenerative diseases with a risk of dementia such as Alzheimer's disease, Lewy body dementia, and Parkinson's disease. Obtaining valid epidemiological data is nevertheless quite difficult because of methodological problems. As a rule, hallucinations are more prevalent in Lewy body disease than Parkinson's disease or Alzheimer's disease. The prevalence in parkinsonian dementia is about the same as in Lewy body disease. Complex visual hallucinations predominate, auditory or tactile hallucinations are more exceptional. Minor forms (illusions, sensation of presence) are also observed. Recurrence is common, mainly in the evening or at night. Patients with advanced mental impairment generally take the hallucinations for reality. The hallucinations can be associated with psychological and behavioral disorders such as delusionnal idea or identification disorders. It is important to search for other causes of hallucinations such as drugs, ocular disorders, or depression, but many of these disorders are common comorbidities in elderly patients with degenerative disease. There is no unique model fitting all the hypothesized pathogenic mechanisms. Complex visual hallucinations most likely arise from abnormal activation of the extra-striat temporal associative regions, but only hypothetical mechanisms have been proposed. Genetic studies and functional imaging have not provided convincing evidence. Current focus is placed on an imbalance between deficient cholinergic transmission and preserved or augmented monoaminergic transmission at the cortical level, but other neurotransmission systems could be involved. The dream dysregulation mechanism proposed in Parkinson's disease cannot be generalized. The link between cognitive disorders and hallucination is also poorly understood: hallucinations are associated with more severe cognitive impairments or more rapid cognitive deline in Parkinson's disease and Alzheimer's disease, but the association with specific cognitive disorders remains to be fully explored. PMID- 15118552 TI - [Brain substrates of episodic memory disorders in Alzheimer's disease]. AB - Resting state PET measurement is useful to unravel brain regions whose dysfunction is responsible for impairment of episodic memory in Alzheimer's disease. First, the consistent hypometabolism of posterior cingulate cortex, temporo-parietal cortex and frontal cortex contrasts with the frequent lack of hippocampal hypometabolism, although it is first to be concerned by neurofibrillary tangles. Several hypotheses are proposed to explain this paradoxical result. Second, the correlative approach (correlations between memory performances and metabolic values on a voxel basis) shows that dysfunction of the hippocampal region is responsible for the earliest deficits of episodic memory, and then suggests the recruitment of neocortical temporal areas normally involved in semantic memory, perhaps as a form of a compensatory mechanism. When applied to the study of Mild Cognitive Impairment, this approach is also very fruitful. PMID- 15118553 TI - [At the boundary between normal aging and Alzheimer disease]. AB - Accurately predicting the development of probable Alzheimer's disease (AD) early in the course of the disease would have major implications. Hence, the pre dementia stage of AD has become a major topic of current research. Amnestic 'mild cognitive impairment' (MCI) has recently emerged as the most convenient avenue to address this Issue, since most of MCI patients will progress to AD, at a rate of 10% to 15% per Year, suggesting that MCI represents the clinical manifestation of incipient AD (Petersen et al., 2001). However, all MCI patients would probably not convert to AD, at least in the near future, so that the Issue of prospective longitudinal studies is to detect specific indices of rapid conversion. In this paper, we focus on longitudinal studies using either neuropsychology, or morphological or functional neuroimaging to find predictive markers allowing to distinguish those MCI patients that rapidly convert to AD from those that do not develop the disease during the follow-up period. Whereas functional neuroimaging, and more specifically FDG-PET, seems particularly accurate to predict AD, the combination of multiple approaches is likely to be a promising avenue. PMID- 15118554 TI - [Neuropsychological rehabilitation in early stage Alzheimer's disease: principles, methods and perspectives]. AB - An increasing number of studies have demonstrated the efficiency of various neuropsychological interventions on individuals with early Alzheimer's disease. There are also some encouraging findings suggesting that the interventions can be successfully applied in less controlled real-world contexts and largely provided into caregiving systems. The interventions have to be directly focused on daily life situations and they should include the perspective of the patients by allowing them to retain a capacity to maximize their cognitive abilities, to state specific preferences and needs, and to make intervention-related decisions. PMID- 15118573 TI - Side effects of medical therapy for chronic hepatitis C. AB - The treatment of chronic hepatitis C virus (HCV) has improved greatly over the past decade. Over half of all patients treated with the combination of peginterferon and ribavirin have the opportunity to achieve sustained virologic response. The major factors which interfere with this goal are the side effects of therapy which require that the doses of peginterferon or ribavirin be reduced or that these medications be discontinued. While some of these side effects can be overcome and treatment continued, some side effects are severe and potentially life threatening. Appropriate recognition and management of these side effects will both improve response to therapy and avoid unnecessary morbidity and mortality. PMID- 15118574 TI - Co-regulation of expression of phase II metabolizing enzymes and multidrug resistance-associated protein 2. AB - Treatment of experimental animals with prototypical enzyme inducers represents a useful tool to characterize the role of different isozymes in drug metabolism and to improve our knowledge on factors regulating their synthesis at the transcriptional level. The effect of model enzyme inducers on phase II (conjugating) enzyme families, including UDP-glucuronosyltransferase's and glutathione-S-transferase's, has been well characterized in rodent liver. More recently, the effect of inducers on the expression of canalicular multidrug resistance-associated protein 2 (Mrp2) has been focused upon. The identification of a number of conjugated drugs as Mrp2 substrates suggests that both the conjugation and transport systems act coordinately to improve drug elimination from the body. We provide evidence about circumstances resulting in the simultaneous upregulation of phase II enzymes and Mrp2 in hepatic and extrahepatic tissues, most likely involving activation of common nuclear receptors (e.g. FXR, PXR). Additionally, we provide an analysis of examples of drug-induced toxicity leading to the simultaneous downregulation of both systems. Potential therapeutic strategies based on the modulation of expression of these systems are also briefly commented upon. PMID- 15118575 TI - A concise and structured review of drug-induced toxic hepatic disease. PMID- 15118576 TI - Changes of coagulation inhibitors and fibrinolysis system in newborn infants with transitory neonatal cholestasis. AB - INTRODUCTION: The hemostatic system in newborn is a dynamic evolving process health-status dependent. OBJECTIVE: To explore the changes in coagulation inhibitors and fibrinolytic system in newborn with neonatal cholestasis, according liver damage intensity. METHODS: In a cross-sectional design, we studied fibrinolysis and coagulation inhibitor proteins, and serum ferritin (SF) in patients with neonatal cholestasis. We stratified the cases according the results of ALT or AST < 100 UI (group I) and > or =100 U/L (group II). RESULTS: We included 24 newborn, 8 for Group I and 16 cases for group II. We documented statistical differences in ATIII values (43.4 vs 27.4%, p < 0,01), plasmin inhibitor (93.5 vs 63.6%, p < 0.01), SF (649 vs 1410 ug/L, p 0.01). The group II cases showed association with SF values (f 20.6, p < 0.01) and plasmin inhibitor (f 40.4, p < 0.01). AST and ALT were related significantly to ATIII concentrations and SF. DISCUSSION: We documented tendency to prothrombotic state (lower ATIII and greater plasmin inhibitor activity), with low plasminogen related to the intensity of liver dysfunction in neonatal cholestasis. We need to determine the role of iron overload in physiopathology of the disease. PMID- 15118577 TI - Etiology of liver cirrhosis in Mexico. AB - BACKGROUND: In the last decades it has been suggested that the main cause of liver cirrhosis in Mexico is alcohol. Currently in Western countries hepatitis C virus stage liver disease and liver transplantation. In Mexico, we have no data relative to the etiology of liver cirrhosis. The aim of this study was to investigate the main causes of liver cirrhosis in Mexico. METHODS: Eight hospitals located in different areas of the country were invited to participate in this study. Those hospitals provide health care to different social classes of the country. The inclusion criteria were the presence of either an histological or a clinical and biochemical diagnosis of liver cirrhosis. RESULTS: A total 1,486 cases were included in this study. The etiology of liver cirrhosis was alcohol in 587 (39.5%), HCV 544 (36.6%), cryptogenic 154 (10.4%), PBC 84 (5.7%), HBV 75 (5.0%) and other 42 (2.8%). There was no statistical difference between alcohol and HCV. CONCLUSIONS: We conclude that the main causes of liver cirrhosis in Mexico are alcohol and HCV. PMID- 15118578 TI - Inflammatory pseudotumor of the liver. PMID- 15118580 TI - Surgical rehabilitation of intestine in short bowel syndrome. PMID- 15118579 TI - Symptomatic focal nodular hyperplasia during pregnancy: a case report. AB - A 30-year-old woman presented with hepatomegaly and an audible hepatic bruit at 24 weeks gestation. Non-contrast MRI demonstrated an exophytic 12.6 x 7.8 x 12.8 cm mass arising from the right lobe of the liver with a central scar, suggestive of focal nodular hyperplasia (FNH). Conservative management included monthly abdominal ultrasound examinations until the time of delivery, to assess growth of the mass and monitor for risk of rupture. Seven weeks post partum the patient experienced severe right upper quadrant pain. A CT angiogram of the liver demonstrated a stable mass with no evidence of bleed or rupture and multiple hypervascular masses throughout the liver. Surgical resection of the dominant lesion was performed. Histological examination of the lesion confirmed FNH. The patient is now 22 months post surgery with radiographic evidence of stable multifocal FNH. PMID- 15118581 TI - Skateboards promote world peace. PMID- 15118583 TI - Regulatory T cells generated by the kidney may mediate the beneficial immune effects of combining kidney with heart transplantation. PMID- 15118584 TI - Laparoscopic appendectomy in the elderly. AB - BACKGROUND: Evidence suggests that laparoscopic appendectomy (LA) has advantages over open appendectomy (OA) in the treatment of appendicitis. It remains, however, unclear whether LA is indicated in the elderly patient population. METHODS: Patients with primary International Classification of Diseases, revision 9, procedure codes for LA (n=32406 patients) and OA (n=112884 patients) were selected from the 1998, 1999, and 2000 Nationwide Inpatient Samples. The end points that were under investigation were the length of hospital stay, the rate of routine discharge, and in-hospital morbidity and mortality rates. Multiple linear and logistic regression analyses were performed to assess the risk adjusted association between the surgery type and the patient outcomes. Stratified analyses were performed according to age (65 years and older; less than 65 years old) and to the presence of appendiceal perforation or abscess. RESULTS: After risk adjustment, patients who underwent LA had a significantly shorter mean length of stay (LA, 2.45 days; OA, 3.71 days; P <. 0001), higher rate of routine discharge (odds ratio, 2.80; P <.0001), lower overall complication rate (odds ratio, 0.92; P=.03), and mortality rate (odds ratio, 0.23; P=.001) compared with OA patients. Similar benefits of LA were found in the strata of patients who were less than 65 years old, in elderly patients, and in patients with appendiceal perforation or abscess. CONCLUSION: LA has statistically significant advantages over OA with respect to the length of hospital stay, the rate of routine discharge, and postoperative morbidity and mortality rates for patients who are less than 65 years old, in elderly patients, and in patients with appendiceal abscess or perforation. PMID- 15118585 TI - Health-related quality-of-life in patients with morbid obesity after gastric banding for surgically induced weight loss. AB - BACKGROUND: Physical, emotional, and social functioning are impaired in obesity. It is unknown whether and, if so, to what extent and in which domain obese subjects who lose weight may catch up to normal-weight levels. Our objective was to compare the health-related quality-of-life (HRQL) of obese subjects with that of a normal-weight reference group before and 1 year after a weight loss program that centered around laparoscopic and open gastric banding. METHODS: An HRQL questionnaire consisting of a battery of both generic and specific measures was administered to 50 morbidly obese subjects on 2 occasions and to 100 healthy, normal-weight subjects, matched for age, gender, education, and vocational training. In addition to weight loss and health gain, the influences of achieved weight loss goals, satisfaction with outcome and operative approach (laparoscopy/laparotomy) were assessed. RESULTS: Quality-of-life was significantly impaired in obese subjects. With a substantial weight loss of 35 kg and 42% loss of excessive weight, and correction of disturbed metabolic parameters, they significantly improved in general well-being, health distress, and perceived attractiveness, approaching halfway the values of a normal-weight reference group. Improvement in values for depression and self-regard lagged behind. In physical activity, they bypassed the reference group. Days of sick leave decreased to the level of the reference group. Improvements in HRQL paralleled the rate of weight loss. Personal satisfaction and surgical approach were of minor influence. CONCLUSIONS: The obese subjects' impaired physical and social functioning improved considerably, catching up midway to normal-weight reference values after weight loss. Psychologic amelioration lagged behind. Whether the latter will catch up later and physical/social improvements will be maintained is the subject of further studies. PMID- 15118586 TI - Sentinel lymph node biopsy in rectal cancer--not yet ready for routine clinical use. AB - BACKGROUND: The value of sentinel node biopsy in visceral cancers is uncertain. We evaluated the feasibility and utility of radiocolloid lymphatic mapping and selective lymph node sampling in patients with rectal cancer. METHODS: Forty eight patients with rectal cancer were investigated. Thirty-seven patients had already undergone preoperative radiochemotherapy for locally advanced tumors. Eleven patients underwent primary surgery. An endoscopic injection of 1 mL technetium 99m-sulfur-colloid into the peritumoral submucosa was performed 15 to 17 hours before surgery. Ex vivo identification of the nuclide-enriched "sentinel lymph nodes" (SLNs) was performed using a hand-held gamma-probe. The selected SLNs were then carefully and systematically examined using serial sections and immunohistochemistry. RESULTS: One or more SLNs were found in 46 of the 48 patients. The SLN detection rate was 96%. Sixteen of the 48 patients had lymph node metastases (35%). In 7 of the 16 patients, the SLNs correctly represented the nodal status. In 9 of the 16 patients, the SLN was tumor-free whereas non-SLN harbored metastases. This result represents a sensitivity of only 44%, and a false-negative rate of 56%. Further analysis showed that the method correctly predicted the nodal status only in the small subgroup of 5 patients with early cancer without preoperative radiation. In 4 patients, juxtaregional lymph nodes were excised on the basis of intraoperative radiocolloid detection, leading to upward staging in 1 patient. CONCLUSIONS: Sentinel lymph node biopsy using the radiocolloid technique with ex vivo lymph node identification shows a relatively high detection rate; however, the sensitivity in patients with locally advanced/irradiated rectal cancer is low. Nevertheless, the detection of juxtaregional metastases can improve staging in some patients. Further studies should focus on patients with early rectal cancers where the data were more promising. PMID- 15118588 TI - Single and multiple resections of multiple hepatic metastases of colorectal origin. AB - BACKGROUND: Indications for hepatectomy in patients with 4 or more hepatic colorectal metastases remain controversial. METHODS: A retrospective cohort study was performed with data from 131 patients who underwent a total of 198 hepatectomies. Patients were grouped according to the number of metastases at the initial hepatectomy (analysis 1) or by the total number of metastases removed by multiple hepatectomies (analysis 2). RESULTS: In analysis 1, the risk ratios for death of patients with 4 to 9 and 10 nodules to those with 1 to 3 nodules were 2.12 (95% CI, 0.99-4.23) and 7.32 (95% CI, 2.82-16.9), respectively. In analysis 2, the risk ratios for death were 1.32 (95% CI, 0.66-2.59) and 3.07 (95% CI, 1.41 6.36), respectively. These values in 106 patients with negative surgical margins were 1.52 (95% CI, 0.51-3.73) and 5.40 (95% CI, 1.25-16.5), and 1.06 (95% CI, 0.45-2.32) and 1.70 (95% CI, 0.49-4.61), respectively. In analysis 2, the 5-year survival rates of patients with 1 to 3, 4 to 9, and 10 or more nodules were 51%, 46%, and 25%, respectively. CONCLUSION: Hepatic resection for patients with 4 to 9 nodules clearly is warranted. On the other hand, for patients with 10 or more tumor nodules, surgery cannot be ensured absolutely to be contraindicated in high volume centers at which the surgical mortality rate is nearly zero. PMID- 15118589 TI - Protective effects of exogenous fructose-1,6-biphosphate during small bowel transplantation in rats. AB - BACKGROUND: We assessed the effect of adding exogenous fructose-1,6-biphosphate (F16BP) to the preservation solution (University of Wisconsin storage solution) used during an experimental procedure of small bowel transplantation in rats. METHODS: We studied levels of the nucleotides hypoxanthine/xanthine and adenosine in tissue after cold ischemia, as well as histologic changes and associated deleterious processes such as bacterial translocation produced by the reperfusion associated with the transplantation. RESULTS: The groups of rats treated with F16BP showed the lowest levels of hypoxanthine/xanthine and uric acid, the highest levels of adenosine, and the lowest levels of histologic damage and lactate dehydrogenase release to the bloodstream. Consumption of intestinal hypoxanthine during reperfusion was lowest in the groups treated with F16BP, as was the incidence of bacterial translocation. CONCLUSIONS: This study shows a protective effect of exogenous F16BP added to University of Wisconsin solution during experimental intestinal transplantation in rats. This protective effect, reflected by decreased intestinal damage and bacterial translocation, was related to a decrease in adenosine triphosphate depletion during cold ischemia before intestinal transplantation, and to the reduced availability of xanthine oxidase substrates for free radical generation during reperfusion. PMID- 15118590 TI - Regulation of arginase expression by T-helper II cytokines and isoproterenol. AB - BACKGROUND: Trauma causes a release of catecholamines, transforming growth factor beta (TGF-beta), and T-helper II cytokines (TH2). Individually, these substances also induce arginase in macrophages. The purpose of this study was to determine the synergistic interactions between isoproterenol, TGF-beta, and TH2 cytokines on arginase expression in macrophages. METHODS: Confluent RAW 264.7 macrophages were incubated with various combinations of interleukins 4, 10, and 13 (IL-4, IL 10, IL-13), and TGF-beta with isoproterenol over 48 hours. Arginase activity, as well as arginase I expression by Western blot and reverse transcriptase polymerase chain reaction, were measured. RESULTS: Although isoproterenol, IL-4, IL-10, and IL-13 individually induced arginase, significant synergy between the combination of isoproterenol with either TGF-beta or the TH2 cytokines was observed. All cytokines except IL-10 also induced arginase I protein and mRNA. Arginase II protein was detected in cells exposed to IL-10. CONCLUSIONS: We conclude that isoproterenol synergizes with IL-4, IL-13, and TGF-beta to increase arginase I mRNA and protein, as well as arginase activity in RAW 264.7 macrophages. Further, IL-10 synergizes with isoproterenol to increase arginase activity and arginase II protein. These synergistic mechanisms may compete with nitric oxide synthase for l-arginine substrate, thus shunting away available arginine from nitric oxide production and contributing to cellular immunosuppression observed after trauma. PMID- 15118591 TI - Effects of exogenous ubiquitin in lethal endotoxemia. AB - BACKGROUND: Recent data indicated a potential role for extracellular ubiquitin in hematopoiesis and inflammation. The biological significance and therapeutic potential of these findings in vivo are unknown. Based on its in vitro abilities to inhibit endotoxin-stimulated tumor necrosis factor alpha (TNFalpha) production, we hypothesized that exogenous ubiquitin has salutary effects on sequelae caused by endotoxin in vivo. METHODS: Anesthetized and mechanically ventilated swine were infused with endotoxin for 3 hours. Ubiquitin was administered intravenously either 15 minutes before or 45 minutes after the endotoxin infusion was started. Albumin was administered to a control group. An additional control group received only ubiquitin. Ex vivo endotoxin evoked TNFalpha production was measured using a whole blood assay. Ubiquitin and TNFalpha concentrations were determined by enzyme-linked immunosorbent assay. RESULTS: Ubiquitin reduced mortality (P <.05), prevented development of pulmonary failure (P <.05), reduced fluid requirements (P <.05), and diminished erythema and edema formation. Ubiquitin pretreatment was more effective than treatment 45 minutes after an endotoxin infusion was started. In vivo ubiquitin administration alone inhibited ex vivo endotoxin-evoked TNFalpha secretion, but had no effect on TNFalpha serum levels after endotoxin infusion. CONCLUSION: In vivo ubiquitin administration has salutary actions during lethal endotoxemia and inhibits ex vivo whole blood TNFalpha production upon endotoxin stimulation. The clinical appearance after ubiquitin treatment in endotoxemia indicates the endothelium as another potential target cell population for interactions with ubiquitin. A novel therapeutic approach to a broad variety of diseases, in which endotoxin triggers immune activation, is suggested. PMID- 15118592 TI - Interleukin-1 upregulates anaphylatoxin receptors on mononuclear cells. AB - BACKGROUND: The anaphylatoxins, C3a and C5a, that are generated during trauma, major surgery, or infection are potent proinflammatory mediators that increase interleukin (IL-1) cytokine synthesis. We investigated the effects of IL-1 on anaphylatoxin receptor expression in monocytes. METHODS: A human monocytic cell line, MONO-MAC-6, was used. C3a and C5a binding sites were assayed by competitive binding. Levels of messenger RNA for the C3a and C5a receptors were analyzed by reverse transcriptase-polymerase chain reaction. Changes of free cytosolic Ca(2+) concentration ([Ca(2+)]i) in response to C3a and C5a were measured. RESULTS: Basal MONO-MAC-6 cell sites for C3a and C5a binding were 10900 C3aR/cell (K(d)=2.0 nmol/L), 8700 C5aR/cell (K(d)=0.9 nmol/L). IL-1alpha increased sites for both C3a (61% increase; P <.01) and C5a (71% increase; P <.001). Levels of C3aR and C5aR messenger RNA also increased in IL-1alpha-stimulated cells. Receptors were coupled to functional responses, which were demonstrated by C3a- or C5a-induced [Ca(2+)]i increases. IL-1 receptor antagonist blocked the effects of IL-1alpha upregulation of anaphylatoxin receptors. CONCLUSION: These results suggest that there is an additional link between IL-1 and anaphylatoxins to amplify proinflammatory effects through monocytes and macrophages. Although C3a and C5a can increase the monocyte production of IL-1, IL-1 increases monocyte expression of receptors for these anaphylatoxins, which further amplifies inflammation. PMID- 15118593 TI - Focal adhesion kinase gene silencing promotes anoikis and suppresses metastasis of human pancreatic adenocarcinoma cells. AB - BACKGROUND: Inadequate or inappropriate cell-substrate contact triggers a subset of apoptotic cell death, termed anoikis. Resistance to anoikis is a characteristic of malignant cells that is associated with increased tumorigenesis and metastasis. Focal adhesion kinase (FAK) is an important regulator of cell survival and migration and cell cycle progression. We tested the hypothesis that FAK gene silencing would promote anoikis and reverse acquired anoikis resistance in human pancreatic adenocarcinoma cells. METHODS: FAK expression was assessed by Northern and Western blot analysis. Anoikis was induced in PANC1, BxPC3, MiaPaCa2, and Mia(AR) (an anoikis-resistant derivative of MiaPaCa2) with the use of polyHEMA culture. FAK expression was suppressed by RNA interference. Anoikis was detected by YO-PRO-1/propidium iodide staining and flow cytometry. Fluorometric caspase profiling was performed. Metastasis was assayed in a nude mouse orthotopic xenograft model. RESULTS: The cell lines that were tested showed marked variation in their anoikis resistance, greater resistance being associated with higher levels of FAK expression. FAK gene silencing promoted anoikis in all cell lines and reversed acquired anoikis resistance in Mia(AR), which was associated with increased caspase activation. Suppression of FAK expression also inhibited metastasis in the nude mouse model. CONCLUSION: FAK gene silencing suppresses anoikis resistance in pancreatic adenocarcinoma cells. FAK represents a potential target for novel antimetastatic therapies. PMID- 15118594 TI - Cutaneous lichen amyloidosis in a family with familial medullary thyroid cancer. PMID- 15118603 TI - Newborn screening--should it be part of prenatal care? PMID- 15118604 TI - Antenatal corticosteroids: we continue to learn. PMID- 15118605 TI - Incorporating newborn screening into prenatal care. AB - Research with focus groups of parents suggests that information on newborn metabolic screening needs to be integrated into prenatal care. Such a practice change can only occur with the support of the obstetric community. PMID- 15118606 TI - Choice and dose of corticosteroid for antenatal treatments. AB - Although antenatal glucocorticoids are standard of care for women at risk of preterm delivery before 32 to 34 weeks' gestation, the choice and dosing of the corticosteroid has not been standardized. An analysis of the trial data demonstrates that the risk of neonatal death is decreased with betamethasone, but not dexamethasone. Other clinical data also indicate that betamethasone is the drug of choice for antenatal treatment. The pharmacology of the corticosteroids suggests that a lower total glucocorticoid dose per treatment may be as effective as the current treatment recommendations. However, a change from current practice will require further randomized controlled trials. PMID- 15118607 TI - Is antenatal vitamin K prophylaxis needed for pregnant women taking anticonvulsants? AB - The objective of this cohort study of a consecutive sample of infants exposed during pregnancy to anticonvulsant drugs was to determine if the mother received late pregnancy prophylaxis with vitamin K, and if any infants had signs of hemorrhagic disease. The medical records of 204 neonates exposed to anticonvulsant drugs in utero and 77 unexposed control neonates were retrospectively reviewed. No hemorrhagic disease was observed, and the incidence of bleeding tendencies was not higher in infants exposed to these drugs compared with control infants, despite no prenatal vitamin K supplementation in all but 1 epileptic woman. PMID- 15118609 TI - On the discharge of Hippocratic obligations: challenges and opportunities. PMID- 15118610 TI - Joseph Price oration. The multiple gestation epidemic: the role of the assisted reproductive technologies. AB - Human IVF has transformed so many lives, but there has been one major drawback- namely the so-called epidemic of multiple gestations. PMID- 15118611 TI - Functional role of matrix metalloproteinases in ovarian tumor cell plasticity. AB - OBJECTIVE: We previously demonstrated that aggressive ovarian cancer cells are able to display in vitro vasculogenic mimicry, which is reflected by their ability to form vasculogenic-like networks in 3-dimensional cultures and to express vascular cell-associated markers. The goal of this study was to examine the functional role of specific matrix metalloproteinases in the formation of vasculogenic-like networks and extracellular matrix remodeling in vitro. We also investigated the clinical relevance of matrix metalloproteinase-2 and -9 and membrane type 1-matrix metalloproteinase in human ovarian cancers with evidence of tumor cell-lined vasculature. STUDY DESIGN: Ovarian cancer cells (A2780-PAR, SKOV3, and EG) were seeded onto separate 3-dimensional cultures that contained either Matrigel or type I collagen, in the absence of endothelial cells or fibroblasts. These cultures were treated with either chemically modified tetracycline-3 (general matrix metalloproteinase inhibitor), recombinant tissue inhibitor of metalloproteinase-1 or -2, or function-blocking antibodies to matrix metalloproteinase-2 or -9 or membrane type 1-matrix metalloproteinase. In addition, 78 invasive epithelial ovarian cancers were evaluated for expression of matrix metalloproteinase-2 and -9 and membrane type 1-matrix metalloproteinase and correlated with various clinical parameters. RESULTS: The aggressive ovarian cancer cells (SKOV3 and EG) were able to form in vitro vasculogenic-like networks and contract 3-dimensional collagen I gels, whereas the poorly aggressive A2780 PAR cell line did not. Chemically modified tetracycline-3 completely blocked the network formation. Blocking antibodies to matrix metalloproteinase-2 and membrane type 1-matrix metalloproteinase inhibited the formation of the vasculogenic-like networks and collagen gel contraction, but the antibody to matrix metalloproteinase-9 had no effect on network formation and minimal effect on gel contraction. Treatment of 3-dimensional cultures with tissue inhibitor of metalloproteinase-2 retarded the network formation and only small, partially developed structures were noted that did not form network connections. Tissue inhibitor of metalloproteinase-1 had no appreciable effect on the extent or efficiency of network formation. Human invasive ovarian cancers with evidence of tumor cell-lined vasculature were significantly more likely to have strong epithelial and stromal matrix metalloproteinase-2 and -9 and membrane type 1 matrix metalloproteinase expression (all probability values were <.05). CONCLUSION: Matrix metalloproteinase-2 and membrane type 1-matrix metalloproteinase appear to play a key role in the development of vasculogenic like networks and matrix remodeling by aggressive ovarian cancer cells. Human ovarian cancers with matrix metalloproteinase overexpression are more likely to have tumor cell-lined vasculature. These results may offer new insights for consideration in ovarian cancer treatment strategies. PMID- 15118612 TI - Prediction of optimal versus suboptimal cytoreduction of advanced-stage serous ovarian cancer with the use of microarrays. AB - OBJECTIVE: The purpose of this study was to define gene expression patterns that are associated with the optimal versus suboptimal debulking of advanced-stage serous ovarian cancers. STUDY DESIGN: RNA from 44 advanced serous ovarian cancers (19 optimal, 25 suboptimal) was evaluated with microarrays that contain >22,000 genes. Genes were screened on the basis of their association with debulking status to obtain the top 120 differentially expressed genes. These genes were then used to develop a predictive model for debulking status, which was subjected to out-of-sample cross validation. RESULTS: We found that patterns of expression of 32 genes can distinguish between optimal and suboptimal debulking with 72.7% predictive accuracy. An analysis of the data that were based on clusters of co ordinately expressed genes resulted in only a marginal improvement in predictive accuracy (75%). CONCLUSION: These data support the hypothesis that favorable survival that is associated with optimal debulking of advanced ovarian cancers is due to, at least in part, the underlying biologic characteristics of these cancers. PMID- 15118614 TI - Regulation of expression of tissue plasminogen activator and plasminogen activator inhibitor-1 by dichloroacetic acid in human fibroblasts from normal peritoneum and adhesions. AB - OBJECTIVE: As part of our ongoing studies to understand the biologic mechanisms of wound repair that lead to postoperative adhesions, we have identified characteristics of an adhesion phenotype that differs between fibroblasts that are obtained from human normal peritoneum and adhesions. In this study, we sought to examine whether stimulation of aerobic metabolism would alter differential expression of tissue plasminogen activator and plasminogen activator inhibitor-1, thereby creating a milieu likely to be less favorable to postoperative adhesion development. To examine this issue, we used a compound, dichloroacetic acid, that stimulates the pyruvate dehydrogenase complex, which causes pyruvate to be metabolized in the Kreb's cycle rather than being converted into lactate, thereby switching anaerobic to aerobic metabolism. STUDY DESIGN: Human fibroblasts from normal peritoneum and adhesions were cultured in the absence or presence of dichloroacetic acid (100 microg/mL) for 24 hours, under normal and hypoxic (2% O(2)) conditions. Real-time reverse transcriptase-polymerase chain reaction of tissue plasminogen activator, plasminogen activator inhibitor-1, and a housekeeping gene beta-actin was performed with messenger RNA that was extracted from all treatment points. RESULTS: Dichloroacetic acid stimulated normal peritoneal fibroblast tissue plasminogen activator messenger RNA expression under hypoxic conditions. In adhesion fibroblasts, dichloroacetic acid treatment enhanced tissue plasminogen activator messenger RNA expression under both normoxic and hypoxic conditions. Plasminogen activator inhibitor-1 messenger RNA expression was unaltered by dichloroacetic acid in normoxic normal peritoneal fibroblasts; but during culture under hypoxic conditions, dichloroacetic acid reduced plasminogen activator inhibitor-1 messenger RNA expression. Similarly, in adhesion fibroblasts, dichloroacetic acid reduced plasminogen activator inhibitor 1 messenger RNA expression under both normoxic and hypoxic conditions. As a result, in normal peritoneal fibroblasts under hypoxic conditions and in adhesion fibroblasts under normoxic and hypoxic conditions, dichloroacetic acid greatly increased the tissue plasminogen activator/plasminogen activator inhibitor-1 ratios. CONCLUSION: These findings confirm that fibroblasts from adhesions are characterized by reduced tissue plasminogen activator and increased plasminogen activator inhibitor-1 production. These observations are extended to show the stimulation of oxidative metabolism by dichloroacetic acid increases tissue plasminogen activator expression under hypoxic conditions. Dichloroacetic acid reduces plasminogen activator inhibitor-1 production by hypoxic normal peritoneal fibroblasts and adhesion fibroblasts under hypoxic conditions. The resultant increases in the tissue plasminogen activator/plasminogen activator inhibitor-1 ratios would favor the development of a fibrinolytic milieu, which would be expected potentially to limit postoperative adhesion development. Thus, regulation of metabolic activity of peritoneal cells may provide a target for future interventions for the reduction of the development of postoperative adhesions, particularly as intervention relates to the healing of peritoneal sites that previously had adhesions. (eg, sites of potential adhesion reformation). PMID- 15118616 TI - RAS/RAF mutation and defective DNA mismatch repair in endometrial cancers. AB - OBJECTIVE: Defective DNA mismatch repair is a common genetic abnormality in both colon cancers and endometrial cancers. Cancers with defective DNA mismatch repair have the so-called mutator phenotype and accumulate genetic errors at an increased rate. An early mutational target in cells with defect DNA mismatch repair may be the RAS/RAF pathway. Colon cancers often have KRAS2 mutations and, if not KRAS2 mutations, may have BRAF mutations. This study investigated the spectrum and frequency of mutations in BRAF and KRAS2 in endometrial carcinomas on the basis of mismatch repair status. STUDY DESIGN: Four hundred forty-one patients with endometrial cancer were staged properly and graded and evaluated for mismatch repair status. These patients were then stratified to groups by the degree of microsatellite instability that was observed in their tumors. One hundred forty-six of the selected tumors were then evaluated for KRAS2 and BRAF mutations on the basis of their microsatellite instability. RESULTS: One hundred forty-six endometrioid endometrial cancers were evaluated for KRAS2 and BRAF mutations. Thirty-five cancers (24%) had activating KRAS2 mutations, but only a single BRAF mutation was identified in an microsatellite instability-positive cancer. Twenty-four of 81 microsatellite instability high cancers (29.6%) in which the MLH1 repair gene was methylated had KRAS2 mutations. When compared with the other groups, this finding approached statistical significance (P=.06). KRAS2 mutation status was associated with increasing age at diagnosis (P=.02). CONCLUSION: Despite many similarities between colon and endometrial cancers, the mechanism of the development of endometrial cancers appears to be different from colon cancers in that BRAF is not affected by a mismatch repair problem, because only KRAS2 mutations were seen. In addition, increasing age appears to lead to an increased likelihood that such a mutation will occur. PMID- 15118618 TI - Manipulation of the pill-free interval in oral contraceptive pill users: the effect on follicular suppression. AB - OBJECTIVE: The purpose of this study was to compare follicular suppression that was produced by 3 different oral contraceptive regimens that differ by treatment in the pill-free interval. STUDY DESIGN: In a university setting, 54 women were assigned randomly to receive either 20 microg ethinyl estradiol+100 microg levonorgestrel followed by 7 pill-free days, 20 microg ethinyl estradiol+150 microg desogestrel followed by 2 days of placebo then 10 microg ethinyl estradiol for 5 days, or 28 days of 20 microg ethinyl estradiol plus 150 microg desogestrel. Follicular suppression was evaluated by serial ultrasound scans and by serum and urinary hormone levels during a 2-month study period. Data were analyzed by nonparametric statistical tests. RESULTS: There was a significant difference in follicle count among the 3 groups (P=.005). Women who were treated with a 7-day pill-free interval experienced the least suppression. Estrogen levels were more variable and led to an observation that overweight (body mass index, >25 kg/m(2)) was associated with reduced follicle suppression (relative risk, 1.6; 95% CI, 1.0, 2.7) and higher estrogen levels (relative risk, 5.3; 95% CI, 1.3, 21). CONCLUSION: Contraceptive pill users who were treated with a 7-day pill-free interval demonstrated less follicular suppression than women who were supplemented with either estrogen alone or estrogen plus progestin. Overweight women were less suppressed than women of normal weight. PMID- 15118620 TI - The effects of dexamethasone and betamethasone on surfactant protein-B messenger RNA expression in human type II pneumocytes and human lung adenocarcinoma cells. AB - OBJECTIVE: The purpose of this study was to compare the effect of a single 48 hour exposure to betamethasone or dexamethasone in the NCI-H441 cell line and in human type II pneumocytes. STUDY DESIGN: NCI-H441 cells were exposed 48 hours to varying concentrations of betamethasone or dexamethasone (10(-10) to 10(-7) mol/L) alone or in combination with 1 mmol/L dibutyryl cyclic adenosine monophosphate. Likewise, human type II pneumocytes were exposed 48 hours to varying concentrations of betamethasone or dexamethasone (10(-9) to 10(-7) mol/L) alone or in combination with 1 mmol/L dibutyryl cyclic adenosine monophosphate. The measured outcome was the stimulatory effect on surfactant protein B gene transcription as expressed by surfactant protein B messenger RNA accumulation. The experiment was conducted 5 times in NCI-H441 cells and 6 times in type II cells, in parallel with control. Surfactant protein B messenger RNA was determined at control level and 48 hours after exposure by quantitative reverse transcription-polymerase chain reaction. RESULTS: A similar dose-dependent response in surfactant protein B messenger RNA expression was seen with both betamethasone and dexamethasone. In human type II pneumocytes, the inductive profile of surfactant protein B messenger RNA after 48-hour exposure to betamethasone or dexamethasone was similar to that seen in the NCI-H441 cells. CONCLUSION: Dexamethasone and betamethasone achieved similar dose-response patterns of surfactant protein-B expression in vitro. PMID- 15118622 TI - In utero hematopoietic stem cell transplantation with haploidentical donor adult bone marrow in a canine model. AB - OBJECTIVE: Chimerism can be achieved in a canine model of in utero bone marrow transplantation with > or =1 x 10(8) CD34(+) haploidentical donor cells per kilogram without graft-versus-host disease. STUDY DESIGN: In utero bone marrow transplantation was performed by ultrasound-guided intraperitoneal infusion in 30 to 41-day-old canines with CD34(+) selected cells from paternal bone marrow at doses of 1.3 x 10(8) to 2.5 x 10(10) CD34(+) cells/kg. A method for marking control littermates was developed with intraperitoneal ethiodol. Postnatal studies included histologic, fluorescent in situ hybridization canine Y probe, and polymerase chain reaction-based chimerism analyses. RESULTS: Term survival was 86% to 100% for transplantations > or =34 days versus 14% and 43% at 30 and 31 days. Microchimerism (<1%) was demonstrated in tissues from 4 informative litters that included thymus, liver, skin, spleen, and intestine. Neither gestational age nor donor CD34 cell dosage altered the level of engraftment in these experiments. There was no evidence of graft-versus-host disease. CONCLUSION: In utero bone marrow transplantation in a canine model achieves microchimerism with high CD34(+) cell doses. PMID- 15118624 TI - An unexpected effect of glucocorticoids on stimulation of c-fms proto-oncogene expression in choriocarcinoma cells that express little glucocorticoid receptor. AB - OBJECTIVE: The purpose of this study was to determine the mechanism by which glucocorticoids stimulate c-fms proto-oncogene expression in JAR choriocarcinoma cells, which are reported to lack the glucocorticoid receptor. STUDY DESIGN: Glucocorticoid action on c-fms was tested with the use of ligand binding assays, Northern and Western blotting, immunohistochemistry, quantitative reverse transcriptase-polymerase chain reaction, and nuclear run-off experiments. RESULTS: Dexamethasone stimulated c-fms (EC(50)=1 nmol/L) in JAR cells in a specific manner. Both RU 486 and actinomycin D inhibited dexamethasone stimulation, which suggests receptor-mediated and transcriptionally regulated actions. Neither cytosol or whole cell binding assays nor immunohistochemistry detected glucocorticoid receptor in JAR cells. However, Southern blot analysis of reverse transcriptase-polymerase chain reaction products revealed levels of glucocorticoid receptor messenger RNA in JAR cells that were approximately 100 fold lower than in HeLa control cells. In all but 1 clone among several JAR clones that were tested, there was concordance between presence or absence of glucocorticoid receptor messenger RNA and glucocorticoid sensitivity. CONCLUSION: Some JAR cells contain low levels of glucocorticoid receptor, which mediate dexamethasone stimulation of c-fms expression. Such sensitivity to circulating glucocorticoids confers a survival advantage to these cells by stimulating the c fms-related invasive behavior so characteristic of choriocarcinomas. PMID- 15118626 TI - Accuracy of pelvic examination in the assessment of patients with operable cervical cancer. AB - OBJECTIVE: The purpose of this study was to determine whether pelvic examination identifies factors that suggest the need for radiotherapy after radical hysterectomy for cervical cancer. STUDY DESIGN: This was an observational study that was conducted from July 1, 2000 through December 31, 2002 that comprised 67 patients with stage 1B-2A cervical cancer who underwent primary surgical treatment. Assessments that were made on pelvic examination were compared with pathologic findings. Data were analyzed with the use of descriptive statistics, calculation of sensitivities, specificities, positive and negative predictive values, and determination of odds ratios. RESULTS: The overall spectrum of small to large tumors (<1-8 cm) and cervices (2.5-8 cm) correlated well with examination (r=0.77-0.88). The accuracy of examination was approximately 50% for tumor diameter (+/-25%), 85% for cervical diameter (+/-25%), 80% for outer-third invasion, 80% for endophytic growth, and 90% for vaginal involvement. The likelihood for adjuvant radiotherapy had a significant association with the number of at-risk examination variables that were present. CONCLUSION: For women who undergo radical hysterectomy for stage 1B to 2A cervical cancer, the presence of multiple high-risk factors that are found on pelvic examination is associated significantly with indications for adjuvant postoperative radiotherapy. PMID- 15118628 TI - Continent ileocolonic urinary reservoir (Miami pouch): the University of Miami experience over 15 years. AB - OBJECTIVE: A patient with a recurrent central pelvic malignancy after radiation will require urinary diversion as part of the reconstructive phase of the pelvic exenteration. The aim of our study was to assess the result of our 15-year experience with a continent ileocolonic urinary reservoir, which is known as the Miami pouch. STUDY DESIGN: Since 1988, all patients who received a continent ileocolonic urinary reservoir in the Division of Gynecologic Oncology, University of Miami School of Medicine, were included in the study. Parameters that were evaluated during the study period include functional outcomes, early and late perioperative complications, and their treatment. RESULTS: A total of 90 patients were identified from February 1988 to December 2002. Seventy-eight patients (87%) had a recurrent central pelvic malignancy, and 82 patients (91%) received radiation before the Miami pouch procedure. The non-reservoir-related morbidities were fever (76%), wound complication (30%), pelvic collection (12%), ileus/small bowel obstruction (12%), and postoperative death (11%). The most common reservoir related complications were urinary infection (40%), ureteral stricture (20%), and difficulty with self-catheterization (18%). In our study, the overall complication rate that was related directly to the Miami pouch was 53%. Conservative treatment resolved>80% of these cases. The rate of urinary continence that was achieved in our patients was 93% during our 15-year experience with the Miami pouch. CONCLUSION: The Miami pouch is a good alternative for continent urinary diversion during exenteration or radiation induced damage. The rate of major complications that require aggressive surgical intervention is acceptable. Most postoperative complications (80%) can be corrected with the use of conservative techniques that are associated with fewer deaths than reoperation and thus should be used first. The technique is simple and effective in women who are at high risk, who have undergone previous radiation therapy, and who have a high rate of functional success and is a profound advantage for a woman's psychosocial well-being. PMID- 15118630 TI - Predictive value of the clinical diagnosis of lower genital tract infection in women. AB - OBJECTIVE: We hypothesized that diagnostic approaches to lower genital tract infections are inaccurate and proposed this study to evaluate typical approaches. STUDY DESIGN: Clinical diagnoses were made with symptoms, direct observation, wet mount, vaginal pH, and amines in 598 women with genital complaints. Laboratory testing for N gonorrhoeae, yeast, T vaginalis, C trachomatis, and bacterial vaginosis by Gram stain. RESULTS: The most frequent symptoms were vaginal discharge (64%), change in discharge (53%), malodor (48%), and pruritus (32%). The infection rates were 46% bacterial vaginosis, 29% yeast, 12% trichomoniasis, 11% chlamydia or gonorrhea; 21% of the patients had no infection. The symptoms did not predict laboratory diagnosis. Clinical signs and symptoms with office based tests and microscopy improved the accuracy of diagnoses. Amsel's clinical diagnosis of bacterial vaginosis was the most sensitive at 92%. The sensitivity of wet mount diagnosis of trichomoniasis was 62%, of yeast by microscopy was 22%, and of mucopus for the prediction of gonorrhea and/or chlamydia was 30%. CONCLUSION: Symptoms alone should not be used to direct treatment in instances in which resources permit more complete evaluation with office-based testing that includes microscopy. Treatment failures or diagnostic uncertainty should prompt specific laboratory testing. PMID- 15118632 TI - The obstetric vesicovaginal fistula: characteristics of 899 patients from Jos, Nigeria. AB - OBJECTIVE: The purpose of this study was to describe the characteristics of women with obstetric vesicovaginal fistulas at a hospital in north central Nigeria. STUDY DESIGN: A retrospective record review was conducted of all women who were seen with vesicovaginal fistulas at Evangel Hospital in Jos, Plateau State, Nigeria, between January 1992 and June 1999. RESULTS: A total of 932 fistula cases were identified, of which 899 cases (96.5%) were associated temporally with labor and delivery. The "typical patient" was small and short (44 kg and <150 cm); had been married early (15.5 years) but was now divorced or separated; was uneducated, poor, and from a rural area; had developed her fistula as a primigravida during a labor that lasted at least 2 days and which resulted in a stillborn fetus. CONCLUSION: Obstetric vesicovaginal fistula is extremely common in north central Nigeria. A complex interaction that involves multiple biologic and socioeconomic factors appears to predispose young women to this devastating childbirth injury. PMID- 15118634 TI - Long-term follow-up after conservative surgery for rectovaginal endometriosis. AB - OBJECTIVE: The purpose of this study was to evaluate long-term results in patients who received conservative surgical treatment for rectovaginal endometriosis. STUDY DESIGN: We analyzed the follow-up data for 83 women who underwent surgery for rectovaginal endometriosis. The inclusion criteria were age 20 to 42 years, moderate-to-severe pain symptoms, conservative treatment with retention of the uterus, and at least 1 ovary; the follow-up period was > or =12 months. Kaplan-Meier analysis and Cox regression were used to calculate recurrence rates. RESULTS: The cumulative rates of pain recurrence, clinical or sonographic recurrence, and new treatment were 28%, 34%, and 27%, respectively. The younger patients had the higher risk of recurrence. Pregnancy had protective effects against the recurrence of symptoms and a need for a new treatment. Patients who underwent bowel resection had fewer recurrences. CONCLUSION: Segmental resection and anastomosis of the bowel, when necessary, improves the outcome without affecting chances of conception. Higher recurrence rates in younger patients seems to justify a more radical treatment in this group of women. PMID- 15118635 TI - Patient satisfaction and changes in prolapse and urinary symptoms in women who were fitted successfully with a pessary for pelvic organ prolapse. AB - OBJECTIVE: Our purpose was to estimate, in women after 2 months of pessary use, patient satisfaction and the percentage of prolapse and urinary symptoms that improve or worsen. STUDY DESIGN: In a prospective study, 100 consecutive women with symptomatic pelvic organ prolapse were fitted with a pessary, and 73 women had a successful 2-week pessary fitting trial. Prolapse and urinary symptoms were assessed at baseline and at 2 months. Patient satisfaction was assessed at 2 months. Risk factors for patient dissatisfaction were assessed. RESULTS: Nearly all prolapse symptoms resolved from baseline to 2 months, respectively: bulge (90% to 3%; P<.001), pressure (49% to 3%; P<.001), discharge (12% to 0%; P=.003), and splinting (14% to 0%; P=.001). Among women with concurrent urinary symptoms at baseline, stress incontinence improved in 45%, urge incontinence improved in 46%, and voiding difficulty improved in 53%, after 2 months. However, among women without urinary symptoms at baseline, occult (de novo) stress incontinence occurred in 21%, de novo urge incontinence occurred in 6%, and de novo voiding difficulty occurred in 4%. At 2 months, 92% of the women were satisfied with their pessary. Six women (8%) were dissatisfied and discontinued use of the pessary. Dissatisfaction was associated with occult stress incontinence (odds ratio, 17.1; 95% CI, 1.9, 206; P=.004). CONCLUSION: After 2 months, 92% of women with a successful pessary fitting trial were satisfied. Nearly all prolapse symptoms resolved after 2 months; 50% of urinary symptoms improved, but occult stress incontinence was a common side effect. PMID- 15118636 TI - Does Burch colposuspension cure coital incontinence? AB - OBJECTIVE: The purpose of this study was to evaluate the effect of Burch colposuspension for stress urinary incontinence on concomitant coital incontinence. STUDY DESIGN: The urogynecology database was searched for sexually active women, who experienced coital incontinence on vaginal penetration, orgasm, or both and who had subsequently undergone Burch colposuspension for urodynamic stress incontinence. The women were interviewed or sent a questionnaire on postoperative bladder and sexual function after a minimum follow-up time of 6 months. RESULTS: Thirty of 43 women answered the questionnaire. Preoperatively, 22 women (73%) experienced urinary leakage during penetration, 3 (10%) during orgasm and 5 (17%) at both. Stress incontinence symptoms were successfully treated in 23 (77%). Coital incontinence was cured in 21 of 30 (70%) and improved in 2. CONCLUSION: The results of this small series suggest that coital incontinence is likely to be cured or improved when stress incontinence has been successfully treated by Burch colposuspension. PMID- 15118637 TI - A comparison of anticipatory and postprocedure pain perception in patients who undergo urodynamic procedures. AB - OBJECTIVE: The purpose of this study was to compare anticipatory and postprocedure pain perception in female patients who undergo multichannel urodynamic evaluation in an office setting. STUDY DESIGN: One hundred consecutive patients completed a visual analogue pain scale before and after urodynamic testing. RESULTS: The mean postprocedure pain score of 2.32 cm was significantly lower than the anticipatory pain rating of 4.35 cm (P<.05). The lower postprocedure pain score was not influenced by previous hysterectomy, body mass index, menopausal status, estrogen replacement therapy, or analgesic or psychiatric medication usage. Patients who had undergone previous anti incontinence surgery reported significantly higher levels of pain during the procedure (mean visual analogue pain scale score, 3.10 cm vs 2.06 cm; P=.027). CONCLUSION: Patients who undergo urodynamic testing anticipate higher degrees of discomfort than they perceive during the procedure. Previous anti-incontinence surgery appears to lower the pain threshold. PMID- 15118638 TI - Comparison between primary peritoneal and epithelial ovarian carcinoma: a population-based study. AB - OBJECTIVE: This study was undertaken to characterize primary peritoneal carcinoma (PPC) compared with ovarian carcinoma (OvC). STUDY DESIGN: Within the framework of a nationwide epidemiologic Israeli study, 95 PPC patients were identified and compared with 117 FIGO stage III-IV epithelial OvC patients matched by age and continent of birth. Data were abstracted from medical records and personal interviews. RESULTS: Our data confirm the similarities between PPC and OvC. A higher rate of abdominal distention, volume of ascites, and malignant cells in ascitic fluid and lower rate of pelvic palpable mass and personal breast cancer history were found in the PPC compared with the OvC group. The overall survival was similar in both groups (30-33 months). In optimally cytoreduced patients, survival was better in the OvC group. Diameter of residual disease was associated with better survival only in the OvC group. CONCLUSION: The clinical differences do not enable a preoperative distinction between the neoplasms. PMID- 15118639 TI - Usefulness of human papillomavirus testing in the follow-up of patients with high grade cervical intraepithelial neoplasia after conization. AB - OBJECTIVE: We aimed to define the adjunctive role of human papillomavirus (HPV) DNA testing in the follow-up of high-grade cervical intraepithelial neoplasia (CIN) after conization. STUDY DESIGN: We analyzed a consecutive series of 2,154 patients who received conization. Patients who had cone diagnosis of cervical cancer or CIN 1, a hysterectomy within 12 weeks after conization, and no follow up data were excluded. The remaining 765 patients (monitored by Pap smears, colposcopy with or without high-risk HPV DNA testing) were analyzed. RESULTS: Of the 765 patients, 279 had CIN at cone margin or endocervix (group A) while 486 were both margin- and endocervix-free (group B). The 3-year cumulative rate of residual/recurrent high-grade CIN was 10.3% (95% CI, 6.9-13.7). HPV follow-up status (P=.015), margin status (P=.001), and follow-up cervical cytology (P<.0001) were significant predictors for residual/recurrent high-grade CIN by multivariate analysis. Four high-grade CINs and 1 microinvasive carcinoma of group A were detected initially by HPV testing, while 48.3% (199/410) of those without recurrent/persistent high-grade CIN still had persistent HPV infection. CONCLUSION: HPV DNA testing is useful in the follow-up and understanding of the natural history after conization for high-grade CIN. PMID- 15118641 TI - Sex chromosomal mosaicism in the gonads of patients with gonadal dysgenesis, but normal female or male karyotypes in lymphocytes. AB - OBJECTIVES: In most cases, XX or XY gonadal dysgenesis remains genetically unexplained. In this pilot study we searched for sex-chromosomal mosaicism in gonads of patients with XX or XY gonadal dysgenesis of undetermined origin. STUDY DESIGN: Gonadal tissues were analyzed by cytogenetic and interphase fluorescence in-situ hybridization (FISH) analyses in four patients with gonadal dysgenesis and normal female (46,XX) or male (46,XY) karyotypes in lymphocytes. RESULTS: Cytogenetic and FISH analyses of the gonads demonstrated in three patients a sex chromosomal mosaicism. Cytogenetic analysis of gonadal tissue of the fourth patient confirmed the result of the lymphocytes with 46,XX, but FISH analysis revealed in 17% of nuclei only one X-chromosome. CONCLUSION: Our data indicate that sex-chromosomal mosaicism in gonads may be a frequent cause of gonadal dysgenesis despite of normal karyotypes in lymphocytes. Therefore, cytogenetic and FISH analyses of gonadal tissue can provide important information in unexplained cases of gonadal dysgenesis. PMID- 15118640 TI - Cardiovascular effects of 6 months of hormone replacement therapy versus placebo: differences associated with years since menopause. AB - OBJECTIVE: In response to post-Women's Health Initiative dialog regarding individualized hormone replacement therapy (HRT), this study evaluates cardiovascular and neuroendocrine effects of HRT versus placebo in postmenopausal women grouped according to time since menopause. STUDY DESIGN: Resting and stress blood pressure (BP), hemodynamic, plasma catecholamine, and cholesterol levels were obtained in 69 women randomly assigned to placebo or active HRT in a 6-month double-blind study. Analyses evaluated if treatment effects differed among those postmenopausal less than 5 years versus 5 years or more. RESULTS: Compared with the placebo-treated and HRT > or =5 groups, the HRT < 5 group showed reduced BP (P<.0007) and trends toward reduced vascular resistance and norepinephrine (P<.07). HRT > or =5 group generally did not differ from placebo. CONCLUSION: Reduced BP and sympathetic tone are evident in some HRT users, with diminishing benefit after the initial postmenopausal years. Time since menopause may be an important consideration in making individualized patient treatment decisions. PMID- 15118643 TI - Patient perceptions of breast biopsy procedures for screen-detected lesions. AB - OBJECTIVE: This study was undertaken to compare patient perceptions of 2 common image-guided breast biopsy procedures on 3 main outcomes: decision making about which procedure to undergo, its convenience, and its side effects. METHODS: Women who had either an excisional or ultrasound-guided core needle breast biopsy in 1997 for a screen-detected lesion had telephone interviews 1 to 3 months after the biopsy. Bivariate associations were tested by using chi(2) and t test statistics. Mulitvariate analyses were used to control for effects of demographic characteristics. RESULTS: Most women (66%) could not remember being offered a choice of procedures, and of those who did have a choice, a higher proportion had an excisional biopsy. Only 2% reported being told the cost of the biopsy procedure. Women who had an excisional biopsy compared with those who had undergone a core needle biopsy reported statistically more hours and days off from work and reported more side effects 1 to 3 days after the biopsy (P<.05). Associations between side effects and type of biopsy procedure were unchanged when adjustment was made for demographic characteristics. CONCLUSION: Women who had the ultrasound-guided needle biopsy reported significantly fewer side effects and needed less time off from work. When a suspicious lesion is noticed on a screening mammogram, it is important that women and their physicians discuss the benefits and risks of the various biopsy procedures before deciding how to proceed, allowing for informed choice. PMID- 15118644 TI - Tetanus and diphtheria immunity among females in the United States: are recommendations being followed? AB - OBJECTIVE: The purpose of this study was to examine prevalence and factors associated with tetanus and diphtheria immunity among women in the United States. STUDY DESIGN: Sera from 9411 female participants from the third National Health and Nutrition Examination Survey were tested for diphtheria and tetanus antitoxin. Interview information for adult women was analyzed to examine associations with immunity. RESULTS: Fifty-seven percent of the female subjects who were > or =6 years old were positive for diphtheria, and 64% of the female subjects for tetanus anti-toxin. Among women > or =20 years old, only 41% of the women were protected against both antigens. Older age, birth outside the United States, and less education was associated with lower immunity. Markers for contact with the health care system were not related to higher immunity. CONCLUSION: More than one half of US women > or =20 years old who were tested were not protected fully against diphtheria and tetanus. All physicians, including obstetricians and gynecologists who may be the sole medical providers for women, should be familiar with the current Advisory Committee on Immunization Practices recommendations regarding tetanus and diphtheria toxoid booster vaccines. PMID- 15118645 TI - First trimester ultrasound screening is effective in reducing postterm labor induction rates: a randomized controlled trial. AB - OBJECTIVE: This study was designed to test the null hypothesis that first trimester ultrasound crown-rump length measurement for gestational age determination will result in no difference in the rate of induction of labor for postterm pregnancy, compared with second trimester biometry alone. STUDY DESIGN: Two hundred eighteen women were randomly assigned to receive either first trimester ultrasound screening or second trimester ultrasound screening to establish the expected date of confinement. Sample size was calculated by using a 2-tailed alpha=.05 and power (1-beta)=80%. Data were analyzed with chi(2) and Fisher exact tests. RESULTS: Of 104 women randomly assigned to the first trimester screening group, 41.3% had their gestational age adjusted on the basis of the crown-rump length measurement. Of 92 women randomly assigned to the second trimester screening group, 10.9% were corrected as a result of biometry (P <.001, relative risk=0.26, 95% CI=0.15-0.46). Five women in the first trimester screening group and 12 women in the second trimester screening group had labor induced for postterm pregnancy (P=0.04, relative risk=0.37, 95% CI=0.14-0.96). CONCLUSION: The application of a program of first trimester ultrasound screening to a low-risk obstetric population results in a significant reduction in the rate of labor induction for postterm pregnancy. PMID- 15118646 TI - Experimental intrauterine infection with Prevotella bivia in New Zealand White rabbits. AB - OBJECTIVE: The purpose of this study was to develop a model of chronic intrauterine and fetal infection with Prevotella bivia, an anaerobe of the lower genital tract that is associated often with bacterial vaginosis. STUDY DESIGN: Thirty timed pregnant New Zealand White rabbits on gestational day 21 were inoculated with P bivia or saline solution in a planned ratio of 4:1 (24 P bivia: 6 saline solution). Rabbits were inoculated 6 cm transcervically with 10(5) to 10(8) colony-forming units/uterine horn of P bivia or with saline solution. Necropsy was scheduled on days 4, 6, or 7 after inoculation. Cultures were collected from blood, uterus, amniotic fluid and fetal brain, lung, and heart. Tissues from placenta, uterus, fetal brain, and lung were evaluated with the histologic inflammation score, with a range of 0 to 13. Amniotic fluid was assayed for tumor necrosis factor-alpha by bioassay. Animals with contamination by other organisms were excluded. Categoric data were evaluated with the use of the Fisher exact test, and continuous data were evaluated with the use of the Wilcoxon rank sum. RESULTS: After the exclusion of 8 animals because of contamination with other organisms, 22 animals were evaluated. Of 3 rabbits with an inoculum of 10(8) P bivia colony-forming units/horn, 2 animals (67%) had fever within 24 hours. These results were not compatible with chronic, subclinical infection. Therefore, 14 does had inocula of 10(5-6) P bivia colony-forming units/horn, with necropsy planned at day 4 (n=5 animals), day 6 (n=3 animals), and day 7 (n=6 animals), and 5 animals were inoculated with saline solution. Animals that had been inoculated with P bivia were significantly more likely to have a positive culture than were those animals that were inoculated with saline solution (64% vs 0%; P<.04). Preterm delivery without fever occurred in 21% of does (3/14 does) that were inoculated with P bivia overall and in 33% of the does (3/9 does) that were followed for 6 to 7 days. No saline-solution inoculated animal had preterm birth. There was an increase in amniotic fluid tumor necrosis factor-alpha levels over time in the P bivia group (P=.12). Histologic inflammation scores were not significantly different between P bivia and saline solution groups. CONCLUSION: Inoculation with P bivia at 10(5-6) colony-forming units/horn leads to chronic intrauterine and fetal infection that are accompanied by preterm birth in up to 33% of cases. This model may serve to explore the mechanism of preterm birth that is induced by chronic infection with genital tract anaerobes. PMID- 15118647 TI - Cell-free fetal DNA in the cerebrospinal fluid of women during the peripartum period. AB - OBJECTIVE: The purpose of this study was to determine whether cell-free fetal DNA is detectable in the cerebrospinal fluid of women during pregnancy and after delivery. STUDY DESIGN: Cerebrospinal fluid was collected from 39 women who underwent an indicated spinal anesthesia procedure. Twenty-six samples were from women who carried at least 1 male fetus, and 13 samples were from women with only a female fetus. DNA was analyzed with the use of real-time polymerase chain reaction for DYS-1 (which represented male fetal DNA) and beta-globin (which represented maternal and fetal DNA). RESULTS: beta-Globin DNA was detected in all cerebrospinal samples. DYS-1 gene sequences were detected in 4 cerebrospinal fluid samples from women who had male fetuses (2 samples were from women who underwent cesarean delivery of singleton pregnancies, 1 sample was from a triplet pregnancy, and 1 sample was from a woman after delivery). No male DNA was detected in the cerebrospinal fluid of women who carried female fetuses. CONCLUSION: Male fetal cells and/or cell-free fetal DNA is detectable in the cerebrospinal fluid of some pregnant women or some women after delivery. PMID- 15118648 TI - Obesity, obstetric complications and cesarean delivery rate--a population-based screening study. AB - OBJECTIVE: This study was undertaken to determine whether obesity is associated with obstetric complications and cesarean delivery. METHODS: A large prospective multicenter database was studied. Subjects were divided into 3 groups: body mass index (BMI) less than 30 (control), 30 to 34.9 (obese), and 35 or greater (morbidly obese). Groups were compared by using univariate and multivariable logistic regression analyses. RESULTS: The study included 16,102 patients: 3,752 control, 1,473 obese, and 877 morbidly obese patients. Obesity and morbid obesity had a statistically significant association with gestational hypertension (odds ratios [ORs] 2.5 and 3.2), preeclampsia (ORs 1.6 and 3.3), gestational diabetes (ORs 2.6 and 4.0), and fetal birth weight greater than 4000 g (ORs 1.7 and 1.9) and greater than 4500 g (ORs 2.0 and 2.4). For nulliparous patients, the cesarean delivery rate was 20.7% for the control group, 33.8% for obese, and 47.4% for morbidly obese patients. CONCLUSION: Obesity is an independent risk factor for adverse obstetric outcome and is significantly associated with an increased cesarean delivery rate. PMID- 15118649 TI - Thrombin and PAR1-activating peptide: effects on human uterine contractility in vitro. AB - OBJECTIVE: Thrombin enhances uterine contractions in animal models, which is an effect that is mediated through protease-activated receptors. The aims of this study were to investigate the effects of thrombin on spontaneous human uterine contractility in vitro, in tissue obtained in the presence and absence of pregnancy, and to investigate the effects of a specific protease-activated receptor 1-activating peptide on human pregnant myometrial contractility. STUDY DESIGN: Isometric recordings were performed under physiologic conditions on myometrial strips that were obtained from elective cesarean delivery and premenopausal hysterectomy specimens. The effects of thrombin (0.5 to 5.0 U/mL) and a specific protease-activated receptor 1-activating peptide (1 nmol/L to 10 micromol/L) on integrals of contractile activity were measured and compared with control values. RESULTS: Thrombin exerted a potent stimulatory effect on human myometrial contractility. For pregnant and nonpregnant myometrium, this effect was significant at concentrations of > or =3.0 U/mL and 1.0 U/mL, respectively, with net maximal stimulatory effects of 44.5% (5.0 U/mL, P<.001) and 40.42% (3.0 U/mL, P<.001), respectively. The protease-activated receptor 1-activating peptide also mediated a significant uterotonic effect (55.1% increase) on human pregnant myometrial contractility at 10 micromol/L concentration (P<.001). CONCLUSION: This uterotonic effect of thrombin suggests that it may play a role in the pathophysiologic condition of human uterine contractions in association with intrauterine bleeding. The similar uterotonic effect that is elicited by protease activated receptor 1-activating peptide suggests a major role for protease activated receptor-1 in the thrombin-mediated contractile effect. PMID- 15118650 TI - Randomized controlled trial of prenatal zinc supplementation and the development of fetal heart rate. AB - OBJECTIVES: This study was undertaken to evaluate whether prenatal zinc supplementation affects maturation of fetal cardiac patterns. STUDY DESIGN: A randomized double-blind controlled trial among 242 low-income Peruvian women was performed. Beginning at 10 to 16 weeks' gestation, women received supplements containing 60 mg iron, 250 microg folic acid with or without 25 mg zinc. Fetal heart rate (mean FHR, variability [HRV], number of accelerations) and movements (number and amplitude of movement bouts, time spent moving) were electronically monitored monthly from 20 weeks' gestation. Developmental trends were evaluated by supplement type among 195 women who completed the trial and had no serious complications of pregnancy. RESULTS: Zinc supplementation was associated with lower FHR, greater number of accelerations, and greater HRV. Supplementation effects on HRV and accelerations were more pronounced after 28 weeks' gestation. No differences in motor activity were observed. CONCLUSION: Prenatal supplementation of zinc-deficient mothers may be beneficial to fetal neurobehavioral development. PMID- 15118651 TI - Trial of labor in patients with a previous cesarean section: does maternal age influence the outcome? AB - OBJECTIVE: This study was undertaken to evaluate the effect of maternal age on the rate of vaginal delivery and the rate of uterine rupture in patients undergoing a trial of labor (TOL) after a prior cesarean delivery. STUDY DESIGN: A cohort study of all women with a live singleton fetus undergoing a TOL after a previous low-transverse cesarean delivery was performed between 1988 and 2002 in a tertiary care center. Patients were divided into 3 groups according to maternal age: less than 30 years old, 30 to 34 years old, and 35 years or older. Women with no prior vaginal delivery and with at least 1 prior vaginal delivery were analyzed separately. The rate of vaginal delivery and the rate of symptomatic uterine rupture were calculated. Multivariate logistic regression analyses were performed to adjust for potential confounding variables. RESULTS: Of the 2493 patients who met the study criteria, there were 1750 women without a prior vaginal delivery (659, 721, and 370, respectively) and 743 women with a prior vaginal delivery (199, 327, and 217, respectively). The rate of uterine rupture was comparable between the groups (2.0%, 1.1%, 1.4%, P=.404 and 0%, 0.3%, 0.9%, P=.312). Successful vaginal delivery was inversely related to maternal age (71.9%, 70.7%, 65.1%, P=.063, and 91.5%, 91.1%, 82.9%, P=.005). After adjusting for confounding variables, maternal age equal to or greater than 35 years old was associated with a lower rate of successful vaginal delivery in patients without prior vaginal delivery (odds ratio [OR] 0.73, 95% CI: 0.56-0.94), and in patients with a prior vaginal delivery (OR: 0.47, 95% CI: 0.29-0.74). CONCLUSION: Patients who are 35 years or older are more prone to have a failed TOL after a prior cesarean delivery. PMID- 15118652 TI - Characteristics of patients seen and services provided in primary care visits in obstetrics/gynecology: data from NAMCS and NHAMCS. AB - Debate continues on the role of obstetricians/gynecologists in women's primary care and there is conflicting data on what these physicians actually do. Using data from nationally representative surveys, we found that obstetricians/gynecologists identify themselves as primary care providers for 21.7% of private physician office visits and 22.2% of hospital-based outpatient clinic visits. Primary care visits more often involved pregnancy care and less often included referred patients. Other demographic characteristics had different effects in community office practices versus hospital outpatient clinic visits. In general, preventive services and counseling were more likely to be provided in general/gynecologic examinations not designated as primary care provider visits. However, the more frequent provision of mental health counseling and lower rates of mammography services in primary care visits suggest that these obstetricians/gynecologists may serve a more longitudinal role in care for their patients. Overall, there continues to be strong variation in the role of obstetricians/gynecologists in women's primary care. PMID- 15118653 TI - Different levels of platelet activation in preeclamptic, normotensive pregnant, and nonpregnant women. AB - OBJECTIVE: The aims of our study were to determine the basal platelet activation state in women with preeclampsia compared with normotensive pregnant women and nonpregnant women and to investigate the platelet reactivity on in vitro stimulation with adenosine diphosphate or thrombin receptor activation peptide. STUDY DESIGN: Platelet expression of CD61 (fibrinogen receptor), CD42a (von Willebrand factor receptor), CD62P (P-selectin), CD63 (Glycoprotein 53), and PAC 1 binding (activated fibrinogen receptor) were determined in 20 pairs of women with preeclampsia/normotensive pregnant women and in 12 nonpregnant women, with the use of flow cytometry. RESULTS: Basal platelet expression of CD61, CD42a and CD62P, and adenosine diphosphate-stimulated CD62P expression were increased in women with preeclampsia compared with normotensive pregnant women. Platelets from women with preeclampsia and normotensive pregnant women differed from platelets from nonpregnant women by expressing higher basal CD63 levels and being more responsive to in vitro agonist stimulation, which was demonstrated by increased expression of CD61, CD62P, and CD63. CONCLUSION: This study supports the notion that platelets are important in the pathophysiologic condition of preeclampsia. PMID- 15118654 TI - Sonographic detection of placenta accreta in the second and third trimesters of pregnancy. AB - OBJECTIVE: The purpose of this study was to determine whether ultrasonography can detect placenta accreta reliably in at-risk patients. STUDY DESIGN: All patients with a previous cesarean delivery and an anterior placenta or placenta previa were evaluated prospectively at each visit for sonographic signs of placenta accreta (interruption of the posterior bladder wall-uterine interface, absence of the retroplacental clear zone, and placental lacunae). RESULTS: This evaluation involved 2002 patients over a 12-year period. Of the 14 patients with a confirmed diagnosis of placenta accreta who had ultrasound examinations between 15 and 20 weeks of gestation, the diagnosis was suspected strongly in 86% of the patients (12/14 patients). There were 18 false-positive cases (54.5%; 18/33 patients), most of which were due to a lack of visualization of the echolucent area between the placenta and the myometrium (obliteration of the 'clear space') during the third trimester. The presence of multiple linear irregular vascular spaces within the placenta (placental lacunae) was the diagnostic sign with the highest positive predictive value for placenta accreta. CONCLUSION: Placenta accreta can be detected as early as 15 to 20 weeks of gestation in most at-risk patients by visualization of irregular vascular spaces within the placenta (placental lacunae). Obliteration of the retroplacental 'clear space' is not a reliable diagnostic sign for placenta accreta. PMID- 15118656 TI - A comparative review of the risks and benefits of hormone replacement therapy regimens. AB - The Women's Health Initiative (a large, randomized, placebo-controlled trial) investigated the effect of conjugated equine estrogens combined with medroxyprogesterone acetate on specific potential long-term benefits and risks. A review of the clinical studies that have investigated different types and regimens of estrogens combined with progestins was conducted to assess how applicable the results of the Women's Health Initiative are to hormone replacement therapy regimens in general. The studies that were reviewed were limited to randomized clinical trials and observational studies that have been published over the last 15 years (1987-2002) and to meta-analyses and reviews that may have included the literature before 1987. The increased risks for venous thromboembolism, stroke, coronary heart disease, and breast cancer that were identified in the Women's Health Initiative trial have also been reported with postmenopausal hormone therapies that contain a variety of estrogen and progestin products. The beneficial effects that were noted in the Women's Health Initiative, with respect to reductions in fractures and colorectal cancer, have not been evaluated in large, randomized controlled trials that use different estrogen/progestin combinations; however, observational trials that used a variety of estrogen or hormone replacement therapy products and randomized clinical studies that evaluated bone mineral density (an excellent predictor of fracture risk) with different estrogen/hormone replacement therapy regimens would suggest that results would be similar to those found in the Women's Health Initiative. Although the relief of menopausal symptoms, the primary reason women seek treatment, was not included in the overall benefit/risk analysis of the Women's Health Initiative, numerous trials suggest that all therapies are effective. Overall, these data indicate that the benefit/risk analysis that was reported in the Women's Health Initiative can be generalized to all postmenopausal hormone replacement therapy products. PMID- 15118657 TI - Evidence-based medicine. PMID- 15118658 TI - [Chitin synthase (CHS) gene analysis of dermatophytes]. AB - About 620-bp genomic DNA fragments of CHS1 genes were amplified from 13 species of dermatophytes by polymerase chain reaction (PCR) and sequenced. The phylogenetic analysis of CHS1 gene fragments of these dermatophyte species revealed that 3 genera of Epidermophyton, Microsporum and Trichophyton were genetically different from each other. The molecular analysis of CHS1 genes will provide useful information for the identification of dermatophytes. The species specific primers were designed from the nucleotide sequences of CHS1 gene in 3 teleomorphs of T. mentagrophytes. Using these primers the PCR analysis identified the clinical isolates of T. mentagrophytes from rabbit as A. benhamiae. By PCR analysis with the dermatophyte specific primer pair, dermatophyte DNA could be diagnosed directly and rapidly in clinical skin samples. The full length of CHS1 and CHS2 genes of Arthroderma benhamiae (one of the teleomorphs of T. mentagrophytes) was sequenced by 5'-RACE and 3'-RACE methods using cDNA as a template. The full length cDNA sequences of CHS1 gene (3158bp) and CHS2 gene (3392bp) were proved to encode 890 and 419 amino acids, respectively. The amino acid sequences of A. benhamiae CHS1 and CHS2 in the conserved regions shared, respectively, about 70% and 80% sequence similarity with those of the other filamentous ascomycetes registered in the data base of the GeneBank. RT-PCR analysis suggested that chitin synthase inhibitors (nikkomycin Z and polyoxin D) might stimulate the expression of CHS1 mRNA in A. benhamiae, and not the expression of CHS2 mRNA. PMID- 15118659 TI - [Molecular taxonomy and identification of pathogenic fungi based on DNA sequence analysis]. AB - Although approximately 80,000 fungi are known, less than 1% are associated with human infection. However, their taxonomy has long been insufficient. During the last decade, DNA sequence analysis was introduced to the taxonomy of pathogenic fungi. Taxonomic advances in the field of medical mycology are helping to identify the causative agents of infectious diseases accurately, facilitating diagnosis and treatment. For example, Malassezia furfur was long considered the major microflora in atopic dermatitis, yet recent studies have indicated that this is not the case, as M. furfur is taxonomically heterogeneous and consists of five species. DNA sequence analysis resolved its taxonomic heterogeneity. Similar examples can be seen in "Candida albicans and C. dubliniensis" and "Trichosporon cutaneum and T. asahii". DNA sequence analysis also enables accurate identification of fungi. At present, almost all pathogenic fungi can be identified by determining the D1/D2 26S rDNA and ITS region of rRNA gene. This paper describes the practical taxonomy and identification of pathogenic fungi based on DNA sequence analysis. PMID- 15118660 TI - [Molecular methods as a diagnostic tool for fungal infections and their prospect]. AB - The current status of molecular biological methods to control mycoses was described. New tools for detecting of specific fungal DNA by reaction at a constant temperature with Loop-mediated isothermal amplification of DNA (LAMP, Eiken Chemical Co., Ltd.), and for strain typing by multilocus sequence typing (MLST) will greatly aid in managing complex fungal infection. PMID- 15118661 TI - [An efficient system for functional hyper-expression of multidrug efflux pumps in Saccharomyces cerevisiae]. AB - Clinically important resistance of fungal pathogens to azole antifungal drugs is most frequently caused by the over-expression of energy-dependent drug efflux pumps. These pumps usually belong to either the ATP-binding cassette (ABC) family or the major facilitator superfamily (MFS) class of membrane transporter. Little is known about how these pumps work and there is urgent need to develop pump antagonists that circumvent resistance. The expression system is based on an S. cerevisiae AD1-8u- strain deleted in seven major ABC transporters which has reduced background and endogenous efflux activity. Plasmid pABC3 was engineered to allow functional hyper-expression of foreign proteins in this host. The main advantages of our system are its cloning efficiency: the use of homologous recombination to stably integrate single copy constructs into the host genome under the control of a highly active transcriptional regulator. The expression system has been used to clone and express genes encoding drug efflux pumps from several pathogenic fungi. Furthermore, functional over-expression of human P glycoprotein was also demonstrated. The protein hyper-expression system will be useful for the screening of pump inhibitors and the study of membrane protein pumping mechanisms. This system has been used to screen chemicals for pump inhibitors. It was found that FK506 and milbemycins chemosensitized pump expressing and fluconazole-resistant strains and inhibited pump ATPase activity. PMID- 15118662 TI - [Molecular cloning of fungal allergens and clinical applications of recombinant allergens in fungal allergy]. AB - A large number of fungi are associated with allergic disorders. There are many problems in using non-standardized fungal extracts for diagnosis of fungal allergy. These problems can be solved by using genetically engineered recombinant allergens. At present, more than 70 fungal allergens have been cloned and sequenced, and the recombinant forms of several of these are commercially available. Measurement of IgE antibodies to these commercially available recombinant allergens could provide the tools useful for characterizing the differential sensitization pattern in relation to a particular disease and the allergenic cross-reactivity among fungi. Only a limited number of recombinant allergens are available at present, thus further studies on the molecular biology of fungal allergens are needed so that more recombinant allergens can be used in the clinical field. PMID- 15118665 TI - [Some problems with clinical trials in the field of dermatology]. PMID- 15118663 TI - [Drug discovery in the new era: exploratory research on novel antifungal agents]. AB - The major antifungal agents currently used in clinics fall into classes of either antibiotics or azoles. Recent introduction of a candin-antibiotic, micafungin, into clinical practice is expected to greatly improve the outcome of therapy in deep mycoses. However, there still exist many mycoses which are hard to treat even with application of a variety of antifungal agents. With this situation of chemotherapy in mycoses, development of novel antifungal agents with good profiles in efficacy and safety and superior to those currently available are anticipated to be discovered by exploratory research. The major target worldwide in the research and development of novel antifungal agents is azole-class compounds. However, among the antifungal antibiotics now being developed, several compounds are being subjected to clinical evaluation based on their novel mechanisms of action and on their non-susceptible feature of cross-resistance to existing antifungal agents. PMID- 15118664 TI - [Status and issues of preclinical evaluation of the therapeutic effects of newly developed antifungal agents]. AB - Preclinical evaluation of the efficacy of an antifungal agent is basically conducted by measuring both the in vitro and the in vivo antifungal activity of the drug using appropriate infection models. Although the first requisite for a method measuring in vitro activity is to obtain results with good reproducibility, an additional requirement is that there be good correlation with the in vivo activity, as described later. For the first condition, in recent years the National Committee for Clinical Laboratory Standards in the United States and the Standardization Committee of the Japanese Society for Medical Mycology have proposed reference techniques with the objective of standardizing drug susceptibility testing; these have been used extensively in measuring antifungal activities of novel agents. However, there are several issues involved when these methods are applied to newly developed drugs. First, standard methods are for particular currently available antifungal agents, but MIC determining standards have not been established for other agents. Reproducibility is therefore not guaranteed. Second, there is a question of whether reliable results can be obtained to test an antifungal spectrum with a limited number of fungal species. On the other hand, in vivo evaluation of novel antifungal agents is extremely important to predict the clinical outcome at the preclinical stage. The important requirements for this in vivo experimental system are: use of an animal model of mycosis that resembles the pathophysiology in humans; use of an administration schedule corresponding to that used in clinical studies, and evaluation of the therapeutic effect considering the dose and administration period. This review presents the present status of preclinical evaluation test methods and discusses the issues. PMID- 15118666 TI - [Strategy for research and development of antifungal agents]. AB - The incidence of deep-seated mycosis has recently been increasing, while the number of clinically available antifungal agents is very limited and each agent has some drawbacks. We focused on the triazole class that is known to have good profiles as antifungal agents. After researching them, we found that CS-758 had excellent profiles in terms of antifungal spectrum, activity, and safety. Until this candidate was obtained, we experienced the following difficulties: (1) In vivo activity did not always reflect in vitro activity. (2) The relationship between in vivo activity and pharmacokinetic profile was important in selecting a candidate as an antifungal agent. (3) Suitable infection-models had to be established to predict clinical efficacy. (4) It was necessary to demonstrate superiority over marketed drugs to develop a novel agent. These experiences give us good ideas for future development of novel antifungal agents. PMID- 15118667 TI - Susceptibility of Pseudallescheria boydii and Scedosporium apiospermum to new antifungal agents. AB - Hyalohyphomycoses caused by Pseudallescheria boydii and Scedosporium apiospermum have recently been on the increase. To find the appropriate treatment for this emerging disease, we examined the antifungal susceptibility of 10 isolates of P. boydii and 17 isolates of S. apiospermum, most of which were isolated from clinical specimens. When the NCCLS M38-P microdilution method was used, itraconazole showed strong antifungal activities, while amphotericin B had little efficacy. A new triazole agent, voriconazole showed a strong effect against isolates of P. boydii and S. apiospermum (MIC50 0.06 micro g/ml), whereas micafungin, a newly developed echinocandin, had little effect (MIC50 >16 micro g/ml). There was no significant difference in the susceptibilities between P. boydii and S. apiospermum isolates against any antifungal agents. Our study suggests that voriconazole is a promising new drug in these infections, and that the same antifungal strategy can be employed in the infections by P. boydii and S. apiospermum. PMID- 15118668 TI - Species identification and strain typing of Fonsecaea pedrosoi using ribosomal RNA gene internal transcribed spacer regions. AB - The restriction fragment length polymorphism (RFLP) in the internal transcribed spacer (ITS) region of the nuclear ribosomal RNA gene (rDNA) was analyzed on Fonsecaea pedrosoi isolates kept in the Department of Dermatology, Kanazawa Medical University, Japan. On the bases of the RFLP patterns with Dde I and Msp I, 131 isolates were classified into 5 types (D1-D5) and 4 types (M1-M4), respectively. Combining the RFLP patterns with Dde I and Msp I, the isolates were further classified into 6 rDNA-types which corresponded to the 6 mtDNA-types reported by Kawasaki et al. based on the mtDNA-RFLP patterns, except for a single strain of mtDNA-type 7, which was indistinguishable from mtDNA-type 2. The strains of each rDNA-type formed a clade on the phylogenetic tree constructed from sequences of the ITS regions. ITS-RFLP analysis discriminated F. pedrosoi from 11 other species of pathogenic phaeoid fungi except F. compacta. These results strongly suggest that the typing based on ITS-RFLP is reliable and that F. pedrosoi and F. compacta are conspecific. Compared with mtDNA-RFLP analysis, ITS-RFLP analysis is less tedious, permits simultaneous analysis of many samples and gives equivalent results rapidly. This analysis is therefore useful for typing or epidemiologically investigating F. pedrosoi and for differentiating it from other dematiaceous fungi. PMID- 15118669 TI - Inductions of germ tube and hyphal formations are controlled by mRNA synthesis inhibitor in Candida albicans. AB - Candida albicans is a pathogenic dimorphic fungus. When yeast cells were pre incubated in YPD medium at 25degreesC and released into HFM7 medium containing 4% serum at 37degreesC, germ tubes emerged within 0.5 h. To determine whether mRNA or protein synthesis was necessary for germ tube formation, we examined the effects of mRNA and protein syntheses inhibitors on this formation. In the presence of cycloheximide, cells were unbudded and no germ tube was observed. However, in the presence of actinomycin D, germ tube formation was observed while budding growth and true hyphae elongation were blocked. Next, we measured mRNA or protein accumulation during induction of germ tube formation in the presence of the inhibitors. In the presence of cycloheximide, protein was not synthesized, while in the presence of actinomycin D, mRNA synthesis decreased to 6.3% and protein synthesis to 37.7%. The condition we found which allows only germination but not budding or filamentation might be convenient to use in screening genes involved in the initial stage of morphological change in C. albicans. PMID- 15118670 TI - No genotype left untreated. PMID- 15118671 TI - The genetic association database. PMID- 15118672 TI - Is mismatch repair really required for ionizing radiation-induced DNA damage signaling? PMID- 15118675 TI - Telomeres in dyskeratosis congenita. PMID- 15118677 TI - Src kinases in Ph+ lymphoblastic leukemia. PMID- 15118676 TI - The spreading influence of chromatin modification. PMID- 15118682 TI - Enlightened enlargement. PMID- 15118683 TI - Robots in space. PMID- 15118684 TI - Young biologists rejected as NIH budget squeezes training grants. PMID- 15118685 TI - Trainee let down as allergy institute withdraws support. PMID- 15118686 TI - Livermore plans ignite protests over nuclear weapons. PMID- 15118687 TI - Cardiologists take heart from stem-cell treatment success. PMID- 15118688 TI - Anthropologists rocked by fossil access row. PMID- 15118689 TI - Gates grant helps Africa develop science academies. PMID- 15118690 TI - Britain seeks compromise on animal research. PMID- 15118691 TI - Victims of extremists join together to change the law. PMID- 15118693 TI - Money changes hands in key bank transaction. PMID- 15118692 TI - Fishy predator gets its teeth into ancient climate history. PMID- 15118695 TI - Art conservation: biology for art's sake. PMID- 15118696 TI - Planetary exploration: a job for the droids? PMID- 15118697 TI - Ten-year review of research in South Africa. PMID- 15118698 TI - Injustice of draft law will speed Italy's brain drain. PMID- 15118699 TI - ITER, fusion for humanity. PMID- 15118700 TI - Dreaming of clean nukes. PMID- 15118706 TI - Learning from the Altmeister. PMID- 15118708 TI - Astrophysics: jump-start for a neutron star. PMID- 15118707 TI - Evolutionary biology: the cod that got away. PMID- 15118709 TI - Hearing: tightrope act. PMID- 15118710 TI - Meteoritics: stars in stones. PMID- 15118712 TI - Zoology: nose of moose. PMID- 15118713 TI - Palaeoanthropology: Neanderthal teeth lined up. PMID- 15118714 TI - Obituary: Sivaramakrishna Chandrasekhar (1930-2004). PMID- 15118716 TI - Animal behaviour: geomagnetic map used in sea-turtle navigation. AB - Migratory animals capable of navigating to a specific destination, and of compensating for an artificial displacement into unfamiliar territory, are thought to have a compass for maintaining their direction of travel and a map sense that enables them to know their location relative to their destination. Compasses are based on environmental cues such as the stars, the Sun, skylight polarization and magnetism, but little is known about the sensory mechanism responsible for the map sense. Here we show that the green sea-turtle (Chelonia mydas) has a map that is at least partly based on geomagnetic cues. PMID- 15118717 TI - Environment: whale-call response to masking boat noise. AB - Background noise can interfere with the detection and discrimination of crucial signals among members of a species. Here we investigate the vocal behaviour in the presence and absence of whale-watcher boat traffic of three social groups (pods) of killer whales (Orcinus orca) living in the nearshore waters of Washington state. We find longer call durations in the presence of boats for all three pods, but only in recent recordings made following a period of increasing boat traffic. This result indicates that these whales adjust their behaviour to compensate for anthropogenic noise once it reaches a threshold level. PMID- 15118718 TI - The path to ubiquitous and low-cost organic electronic appliances on plastic. AB - Organic electronics are beginning to make significant inroads into the commercial world, and if the field continues to progress at its current, rapid pace, electronics based on organic thin-film materials will soon become a mainstay of our technological existence. Already products based on active thin-film organic devices are in the market place, most notably the displays of several mobile electronic appliances. Yet the future holds even greater promise for this technology, with an entirely new generation of ultralow-cost, lightweight and even flexible electronic devices in the offing, which will perform functions traditionally accomplished using much more expensive components based on conventional semiconductor materials such as silicon. PMID- 15118719 TI - The geometry of the double-pulsar system J0737-3039 from systematic intensity variations. AB - Two pulsars (PSR J0737-3039A and B) were recently discovered in highly relativistic orbits around one another. The system contains a rapidly rotating pulsar with a spin period of 22.7 ms and a slow companion with a spin period of 2.77 s, referred to here as 'A' and 'B', respectively. A unique property of the system is that the pulsed radio flux from B increases systematically by almost two orders of magnitude during two short portions of its orbit. Here we report a geometrical model of the system that simultaneously explains the intensity variations of B and provides constraints on the spin axis orientation and emission geometry of A. Our model assumes that B's pulsed radio flux increases when illuminated by emission from A. We predict that A's pulse profile will evolve considerably over the next several years owing to geodetic precession until it disappears entirely in 15-20 years. PMID- 15118720 TI - Stardust silicates from primitive meteorites. AB - Primitive chondritic meteorites contain material (presolar grains), at the level of a few parts per million, that predates the formation of our Solar System. Astronomical observations and the chemical composition of the Sun both suggest that silicates must have been the dominant solids in the protoplanetary disk from which the planets of the Solar System formed, but no presolar silicates have been identified in chondrites. Here we report the in situ discovery of presolar silicate grains 0.1-1 microm in size in the matrices of two primitive carbonaceous chondrites. These grains are highly enriched in 17O (delta17O(SMOW) > 100-400 per thousand ), but have solar silicon isotopic compositions within analytical uncertainties, suggesting an origin in an oxygen-rich red giant or an asymptotic giant branch star. The estimated abundance of these presolar silicates (3-30 parts per million) is higher than reported for other types of presolar grains in meteorites, consistent with their ubiquity in the early Solar System, but is about two orders of magnitude lower than their abundance in anhydrous interplanetary dust particles. This result is best explained by the destruction of silicates during high-temperature processing in the solar nebula. PMID- 15118721 TI - Carbon nanotubes as nanoscale mass conveyors. AB - The development of manipulation tools that are not too 'fat' or too 'sticky' for atomic scale assembly is an important challenge facing nanotechnology. Impressive nanofabrication capabilities have been demonstrated with scanning probe manipulation of atoms and molecules on clean surfaces. However, as fabrication tools, both scanning tunnelling and atomic force microscopes suffer from a loading deficiency: although they can manipulate atoms already present, they cannot efficiently deliver atoms to the work area. Carbon nanotubes, with their hollow cores and large aspect ratios, have been suggested as possible conduits for nanoscale amounts of material. Already much effort has been devoted to the filling of nanotubes and the application of such techniques. Furthermore, carbon nanotubes have been used as probes in scanning probe microscopy. If the atomic placement and manipulation capability already demonstrated by scanning probe microscopy could be combined with a nanotube delivery system, a formidable nanoassembly tool would result. Here we report the achievement of controllable, reversible atomic scale mass transport along carbon nanotubes, using indium metal as the prototype transport species. This transport process has similarities to conventional electromigration, a phenomenon of critical importance to the semiconductor industry. PMID- 15118722 TI - Interdecadal variation in the extent of South Pacific tropical waters during the Younger Dryas event. AB - During the Younger Dryas event, about 12,000 years ago, the Northern Hemisphere cooled by between 2 and 10 degrees C (refs 1, 2) whereas East Antarctica experienced warming. But the spatial signature of the event in the southern mid latitudes and tropics is less well known, as records are sparse and inconclusive. Here we present high-resolution analyses of skeletal Sr/Ca and 18O/16O ratios for a giant fossil Diploastrea heliopora coral that was preserved in growth position on the raised reef terraces of Espiritu Santo Island, Vanuatu, in the southwestern tropical Pacific Ocean. Our data indicate that sea surface temperatures in Vanuatu were on average 4.5 +/- 1.3 degrees C cooler during the Younger Dryas event than today, with a significant interdecadal modulation. The amplified annual cycle of sea surface temperatures, relative to today, indicates that cooling was caused by the compression of tropical waters towards the Equator. The positive correlation in our record between the oxygen isotope ratios of sea water and sea surface temperatures suggests that the South Pacific convergence zone, which brings 18O-depleted precipitation to the area today, was not active during the Younger Dryas period. PMID- 15118723 TI - Polysaccharide aggregation as a potential sink of marine dissolved organic carbon. AB - The formation and sinking of biogenic particles mediate vertical mass fluxes and drive elemental cycling in the ocean. Whereas marine sciences have focused primarily on particle production by phytoplankton growth, particle formation by the assembly of organic macromolecules has almost been neglected. Here we show, by means of a combined experimental and modelling study, that the formation of polysaccharide particles is an important pathway to convert dissolved into particulate organic carbon during phytoplankton blooms, and can be described in terms of aggregation kinetics. Our findings suggest that aggregation processes in the ocean cascade from the molecular scale up to the size of fast-settling particles, and give new insights into the cycling and export of biogeochemical key elements such as carbon, iron and thorium. PMID- 15118724 TI - Maturation trends indicative of rapid evolution preceded the collapse of northern cod. AB - Northern cod, comprising populations of Atlantic cod (Gadus morhua) off southern Labrador and eastern Newfoundland, supported major fisheries for hundreds of years. But in the late 1980s and early 1990s, northern cod underwent one of the worst collapses in the history of fisheries. The Canadian government closed the directed fishing for northern cod in July 1992, but even after a decade-long offshore moratorium, population sizes remain historically low. Here we show that, up until the moratorium, the life history of northern cod continually shifted towards maturation at earlier ages and smaller sizes. Because confounding effects of mortality changes and growth-mediated phenotypic plasticity are accounted for in our analyses, this finding strongly suggests fisheries-induced evolution of maturation patterns in the direction predicted by theory. We propose that fisheries managers could use the method described here as a tool to provide warning signals about changes in life history before more overt evidence of population decline becomes manifest. PMID- 15118725 TI - Surprisingly rapid growth in Neanderthals. AB - Life-history traits correlate closely with dental growth, so differences in dental growth within Homo can enable us to determine how somatic development has evolved and to identify developmental shifts that warrant species-level distinctions. Dental growth can be determined from the speed of enamel formation (or extension rate). We analysed the enamel extension rate in Homo antecessor (8 teeth analysed), Homo heidelbergensis (106), Homo neanderthalensis ('Neanderthals'; 146) and Upper Palaeolithic-Mesolithic Homo sapiens (100). Here we report that Upper Palaeolithic-Mesolithic H. sapiens shared an identical dental development pattern with modern humans, but that H. antecessor and H. heidelbergensis had shorter periods of dental growth. Surprisingly, Neanderthals were characterized by having the shortest period of dental growth. Because dental growth is an excellent indicator of somatic development, our results suggest that Neanderthals developed faster even than their immediate ancestor, H. heidelbergensis. Dental growth became longer and brain size increased from the Plio-Pleistocene in hominid evolution. Neanderthals, despite having a large brain, were characterized by a short period of development. This autapomorphy in growth is an evolutionary reversal, and points strongly to a specific distinction between H. sapiens and H. neanderthalensis. PMID- 15118726 TI - Sperm death and dumping in Drosophila. AB - Mating with more than one male is the norm for females of many species. In addition to generating competition between the ejaculates of different males, multiple mating may allow females to bias sperm use. In Drosophila melanogaster, the last male to inseminate a female sires approximately 80% of subsequent progeny. Both sperm displacement, where resident sperm are removed from storage by the incoming ejaculate of the copulating male, and sperm incapacitation, where incoming seminal fluids supposedly interfere with resident sperm, have been implicated in this pattern of sperm use. But the idea of incapacitation is problematic because there are no known mechanisms by which an individual could damage rival sperm and not their own. Females also influence the process of sperm use, but exactly how is unclear. Here we show that seminal fluids do not kill rival sperm and that any 'incapacitation' is probably due to sperm ageing during sperm storage. We also show that females release stored sperm from the reproductive tract (sperm dumping) after copulation with a second male and that this requires neither incoming sperm nor seminal fluids. Instead, males may cause stored sperm to be dumped or females may differentially eject sperm from the previous mating. PMID- 15118727 TI - Detecting selection using a single genome sequence of M. tuberculosis and P. falciparum. AB - Selective pressures on proteins are usually measured by comparing nucleotide sequences. Here we introduce a method to detect selection on the basis of a single genome sequence. We catalogue the relative strength of selection on each gene in the entire genomes of Mycobacterium tuberculosis and Plasmodium falciparum. Our analysis confirms that most antigens are under strong selection for amino-acid substitutions, particularly the PE/PPE family of putative surface proteins in M. tuberculosis and the EMP1 family of cytoadhering surface proteins in P. falciparum. We also identify many uncharacterized proteins that are under strong selection in each pathogen. We provide a genome-wide analysis of natural selection acting on different stages of an organism's life cycle: genes expressed in the ring stage of P. falciparum are under stronger positive selection than those expressed in other stages of the parasite's life cycle. Our method of estimating selective pressures requires far fewer data than comparative sequence analysis, and it measures selection across an entire genome; the method can readily be applied to a large range of sequenced organisms. PMID- 15118728 TI - Regulation of ethylene gas biosynthesis by the Arabidopsis ETO1 protein. AB - Ethylene gas is used as a hormone by plants, in which it acts as a critical growth regulator. Its synthesis is also rapidly evoked in response to a variety of biotic and abiotic stresses. The Arabidopsis ethylene-overproducer mutants eto2 and eto3 have previously been identified as having mutations in two genes, ACS5 and ACS9, respectively; these encode isozymes of 1-aminocyclopropane-1 carboxylic acid synthase (ACS), which catalyse the rate-limiting step in ethylene biosynthesis. Here we report that another ethylene-overproducer mutation, eto1, is in a gene that negatively regulates ACS activity and ethylene production. The ETO1 protein directly interacts with and inhibits the enzyme activity of full length ACS5 but not of a truncated form of the enzyme, resulting in a marked accumulation of ACS5 protein and ethylene. Overexpression of ETO1 inhibited induction of ethylene production by the plant growth regulator cytokinin, and promoted ACS5 degradation by a proteasome-dependent pathway. ETO1 also interacts with CUL3, a constituent of ubiquitin ligase complexes in which we propose that ETO1 serves as a substrate-specific adaptor protein. ETO1 thus has a dual mechanism, inhibiting ACS enzyme activity and targeting it for protein degradation. This permits rapid modulation of the concentration of ethylene. PMID- 15118731 TI - State of the unions. PMID- 15118729 TI - Splicing of oskar RNA in the nucleus is coupled to its cytoplasmic localization. AB - oskar messenger RNA localization at the posterior pole of the Drosophila oocyte is essential for germline and abdomen formation in the future embryo. The nuclear shuttling proteins Y14/Tsunagi and Mago nashi are required for oskar mRNA localization, and they co-localize with oskar mRNA at the posterior pole of the oocyte. Their human homologues, Y14/RBM8 and Magoh, are core components of the exon-exon junction complex (EJC). The EJC is deposited on mRNAs in a splicing dependent manner, 20-24 nucleotides upstream of exon-exon junctions, independently of the RNA sequence. This indicates a possible role of splicing in oskar mRNA localization, challenging the established notion that the oskar 3' untranslated region (3'UTR) is sufficient for this process. Here we show that splicing at the first exon-exon junction of oskar RNA is essential for oskar mRNA localization at the posterior pole. We revisit the issue of sufficiency of the oskar 3'UTR for posterior localization and show that the localization of unrelated transcripts bearing the oskar 3'UTR is mediated by endogenous oskar mRNA. Our results reveal an important new function for splicing: regulation of messenger ribonucleoprotein complex assembly and organization for mRNA cytoplasmic localization. PMID- 15118732 TI - New lease of life for tropical medicine. PMID- 15118733 TI - Back to where it all started: monoamines and behavior--from drug responses to genes. PMID- 15118736 TI - Complex disorders reloaded: causality, action, reaction, cause and effect. PMID- 15118737 TI - A genome screen of 13 bipolar affective disorder pedigrees provides evidence for susceptibility loci on chromosome 3 as well as chromosomes 9, 13 and 19. PMID- 15118739 TI - Indoor air and exposure: selected papers from INDOOR AIR 2002. PMID- 15118740 TI - Personal, indoor, and outdoor VOC exposures in a probability sample of children. AB - As part of the Minnesota Children's Pesticide Exposure Study we measured volatile organic compound (VOC) concentrations in a probability sample of households with children. The 6-day average concentrations for 10 common VOCs were obtained in urban and nonurban residences twice during this multiphase study: screening-phase indoor measurements were collected in 284 households, and in the intensive-phase matched outdoor (O), indoor (I), and personal (P) measurements were collected in a subset (N=72) of the screened households. Screening-phase households with smokers had significantly higher concentrations of benzene and styrene compared to nonsmoking households; households with an attached garage had significantly higher levels of benzene, chloroform, styrene, and m/p- and o-xylene compared to households without an attached garage; and nonurban residences, which had a greater prevalence of smokers and attached garages, had significantly higher 1,1,1-trichloroethane, styrene, and toluene and significantly lower tetrachloroethylene concentrations compared to urban households. The screening phase weighted distributions estimate the mean and variability in indoor VOC concentrations for more than 45,000 households with children in the census tracts sampled. Overall, median indoor concentrations of most VOCs measured in this study were similar to or lower than indoor levels measured previously in the United States. Intensive-phase outdoor VOC concentrations were generally lower than other major metropolitan areas, but urban concentrations were significantly higher than nonurban concentrations for all compounds except 1,1,1 trichloroethylene. A consistent pattern of P>I>O was observed for nine of 10 VOCs, with 1,1,1-trichloroethylene (I>P>O) being the only exception to this pattern. For most children, the indoor at-home microevironment was strongly associated with personal exposure after controlling for important covariates, but the ratio of median to upper bound exposures was smaller than that observed in studies of adults. There are relatively little data on VOC exposures in children, so these results are useful for estimating the central tendency and distribution of VOC exposures in locations where children spend a majority of their time. PMID- 15118741 TI - Exposure assessment for respirable particulates associated with household fuel use in rural districts of Andhra Pradesh, India. AB - Indoor air pollution associated with combustion of solid fuels seems to be a major contributor to the national burden of disease in India, but relatively few quantitative exposure assessment studies are available. This study quantified the daily average concentrations of respirable particulates (50% cut-off at 4 microm) in 412 rural homes selected through stratified random sampling from three districts of Andhra Pradesh, India and recorded time activity data from 1400 individuals to reconstruct 24-h average exposures. The mean 24-h average concentrations ranged from 73 to 732 microg/m(3) in gas- versus solid fuel-using households, respectively. Concentrations were significantly correlated with fuel type, kitchen type, and fuel quantity. The mean 24-h average exposures ranged from 80 to 573 microg/m(3). Among solid fuel users, the mean 24-h average exposures were the highest for women cooks and were significantly different from men and children. Among women, exposures were the highest in the age group of 15 40 years (most likely to be involved in cooking or helping in cooking), while among men, exposures were highest in the age group of 65-80 years (most likely to be indoors). The data are being used to develop a model to predict quantitative categories of population exposure based on survey information on housing and fuel characteristics. This would facilitate the development of a regional exposure database and enable better estimation of health risks. PMID- 15118742 TI - Impact of improved stoves, house construction and child location on levels of indoor air pollution exposure in young Guatemalan children. AB - The goal of this study was to assess the impact of improved stoves, house ventilation, and child location on levels of indoor air pollution and child exposure in a rural Guatemalan population reliant on wood fuel. The study was a random sample of 204 households with children less than 18 months in a rural village in the western highlands of Guatemala. Socio-economic and household information was obtained by interview and observation. Twenty-four hour carbon monoxide (CO) was used as the primary measure of kitchen pollution and child exposure in all homes, using Gastec diffusion tubes. Twenty-four hour kitchen PM(3.5) was measured in a random sub-sample (n=29) of kitchens with co-located CO tubes. Almost 50% of the homes still used open fires, around 30% used chimney stoves (planchas) mostly from a large donor-funded programme, and the remainder of homes used various combinations including bottled gas and open fires. The 24-h kitchen CO was lowest for homes with self-purchased planchas: mean (95% CI) CO of 3.09 ppm (1.87-4.30) vs. 12.4 ppm (10.2-14.5) for open fires. The same ranking was found for child CO exposure, but with proportionately smaller differentials (P<0.0001). The 24-h kitchen PM(3.5) in the sub-sample showed similar differences (n=24, P<0.05). The predicted child PM for all 203 children (based on a regression model from the sub-sample) was 375 microg/m(3) (270-480) for self purchased planchas and 536 microg/m(3) (488-584) for open fires. Multivariate analysis showed that stove/fuel type was the most important determinant of kitchen CO, with some effect of kitchen volume and eaves. Stove/fuel type was also the key determinant of child CO, with some effect of child position during cooking. The improved stoves in this community have been effective in reducing indoor air pollution and child exposure, although both measures were still high by international standards. Large donor-funded stove programmes need to aim for wider acceptance and uptake by the local families. Better stove maintenance is also required. PMID- 15118743 TI - Elevated personal exposure to particulate matter from human activities in a residence. AB - Continuous laser particle counters collocated with time-integrated filter samplers were used to measure personal, indoor, and outdoor particulate matter (PM) concentrations for a variety of prescribed human activities during a 5-day experimental period in a home in Redwood City, CA, USA. The mean daytime personal exposures to PM(2.5) and PM(5) during prescribed activities were 6 and 17 times, respectively, as high as the pre-activity indoor background concentration. Activities that resulted in the highest exposures of PM(2.5), PM(5), and PM(10) were those that disturbed dust reservoirs on furniture and textiles, such as dry dusting, folding clothes and blankets, and making a bed. The vigor of activity and type of flooring were also important factors for dust resuspension. Personal exposures to PM(2.5) and PM(5) were 1.4 and 1.6 times, respectively, as high as the indoor concentration as measured by a stationary monitor. The ratio of personal exposure to the indoor concentration was a function of both particle size and the distance of the human activity from the stationary indoor monitor. The results demonstrate that a wide variety of indoor human resuspension activities increase human exposure to PM and contribute to the "personal cloud" effect. PMID- 15118744 TI - Comparison of biocontaminant levels associated with hard vs. carpet floors in nonproblem schools: results of a year long study. AB - Choosing the appropriate floor surface for a school environment is a complex issue. To assist school personnel in determining which flooring is best for their school, we studied the biocontaminant levels associated with carpeted and hard surface flooring. Two schools were selected, one predominantly tiled and one predominantly carpeted, as similar as possible with the exception of their floor coverings. Neither school was a "problem" building. Multiple biocontaminants were measured. For flooring, there were statistically significant differences for all the tested biocontaminants except fungi. The carpeted surfaces, being strong sinks, generally had higher surface loadings of the biocontaminants, while the airborne levels were significantly higher over tiled floors. Significant differences in airborne levels were found for dust mass, spores, fungi, beta-1,3 glucans, and endotoxins. The results suggest that carpet flooring was not the major contributor to airborne levels of biocontaminants in these two nonproblem schools. PMID- 15118745 TI - Variability of personal chemical exposure in eight office buildings in Sweden. AB - This study focuses on the variability in chemical exposures for individuals working in office buildings. The study involved eight office buildings with 79 participants, and exposures were measured using personal samplers for volatile organic compounds, aldehydes, amines, nitrogen dioxide, ozone, and particles. Ventilation was assessed in each individual office. "Variability among buildings" and "variability among individuals" were evaluated for any component (of the 123) measured in samples from at least 20 persons, using variance component analysis and principal component analysis. Interpersonal differences explained the major part of the variance for 78% of the compounds versus between-buildings differences for 14% of the compounds. For 8% of compounds, the variation was explained in equal amounts by the differences among individuals and among buildings. This study illustrates the necessity for individualised measurements (versus stationary measurements in building) to estimate personal exposures. These results also support the conclusion that in case-referent studies of "sick building syndrome" (SBS), referents to SBS cases can be randomised for building location. PMID- 15118746 TI - Estimating effects of moisture damage repairs on students' health-a long-term intervention study. AB - Health symptom questionnaire responses were collected from upper secondary and high school students (n=245) before comprehensive repairs of moisture damage in the school. The questionnaire study was repeated 1 year (n=227), 3 years (n=256), and 5 years (n=233) after the repairs. The data were analyzed both in cross sectional design including all respondents, and longitudinally including paired observations of those individuals who had responded both before and after the repairs. In addition, the effect of intervention on health symptoms was analyzed using generalized estimating equations (GEEs), taking into account within-subject correlation between repeated measurements. Compared to the situation before the repairs, the situation after the repairs was significantly improved in most of the 20 symptoms studied among the cross-sectional study populations. However, improvement was not so clear in the paired analysis and GEE analysis among the students who responded to three repeated questionnaires. The results indicate that the repairs succeeded in the sense that new cases of symptomatic students were no longer expected. However, the reversibility of symptoms among the group of exposed individuals may need to be considered separately. PMID- 15118747 TI - Relationship between environmental tobacco smoke and urinary cotinine levels in passive smokers at their residence. AB - Studies of the health effects of environmental tobacco smoke (ETS) using measured air concentrations are subject to bias. Cotinine, a nicotine metabolite detected in urine, has been recommended as a quantitative measure of nicotine intake and thus as a marker for ETS exposure in humans. The aim of this study was to correlate home indoor ETS levels with passive smokers' urinary cotinine levels. The urinary cotinine concentrations of 57 non-smoking women who spend >19 h a day at home and the nicotine levels in their living room air were measured over a period of 24 h. Nicotine and urinary cotinine levels were analyzed using GC/MS and HPLC/UV, respectively. In addition, information was collected regarding the smoking habits of the subjects' families. A significant correlation was found between the nicotine levels in indoor air and the urinary cotinine to creatinine ratio of the passive smokers. The smoking habits of the subjects' family members were also correlated to the urinary cotinine levels of the passive smokers. PMID- 15118748 TI - Inhalation of hazardous air pollutants from environmental tobacco smoke in US residences. AB - In the United States, 48 million adults smoke 3.5-5 x 10(11) cigarettes/year. Many cigarettes are smoked in private residences, causing regular environmental tobacco smoke (ETS) exposure to roughly 31 million nonsmokers (11% of the US population), including 16 million juveniles. (Upper bound estimates are 53 million exposed nonsmokers including 28 million juveniles.) ETS contains many chemical species whose industrial emissions are regulated by the US federal government as hazardous air pollutants (HAPs). In this paper, average daily residential exposures to and intakes of 16 HAPs in ETS are estimated for US nonsmokers who live with smokers. The evaluation is based on material-balance modeling; utilizes published data on smoking habits, demographics, and housing; and incorporates newly reported exposure-relevant emission factors. The ratio of estimated average exposure concentrations to reference concentrations is close to or greater than one for acrolein, acetaldehyde, 1,3-butadiene, and formaldehyde, indicating potential for concern regarding noncancer health effects from chronic exposures. In addition, lifetime cancer risks from residential ETS exposure are estimated to be substantial ( approximately 2-500 per million) for each of five known or probable human carcinogens: acetaldehyde, formaldehyde, benzene, acrylonitrile, and 1,3-butadiene. Cumulative population intakes from residential ETS are compared for six key compounds against ambient sources of exposure. ETS is found to be a dominant source of environmental inhalation intake for acrylonitrile and 1,3-butadiene. It is an important cause of intake for acetaldehyde, acrolein, and formaldehyde, and a significant contributor to intake for benzene. PMID- 15118749 TI - Exposure-response relationships between lifetime exposure to residential coal smoke and respiratory symptoms and illnesses in Chinese children. AB - Data collected in a large epidemiologic study were analyzed to examine respiratory health effects of residential coal use in 7058 school children living in the four Chinese cities of Chongqing, Guangzhou, Lanzhou, and Wuhan. A Scenario Evaluation Approach was used to develop two exposure variables, heating coal smoke and cooking coal smoke. Estimated lifetime exposures to heating coal smoke and cooking coal smoke were both classified into four-level ordinal scales, as follows: no reported exposure (control); lightly exposed; moderately exposed; and heavily exposed. Zero-one dummy variables were constructed for each exposure level other than the control level (total six variables). These variables were entered into the analytical model. We tested for exposure-response relationships using logistic regression models, while controlling for other relevant covariates, including an indicator variable of ambient air pollution levels. We observed monotonic and positive exposure-response relationships of exposure to heating coal smoke with modeled odds ratios (ORs) of phlegm, cough with phlegm, and bronchitis. Other health outcomes were not associated with such exposure in a monotonic exposure-response pattern. However, ORs for cough, wheeze, and asthma were all higher in the exposed groups than in the control group. We observed no consistent associations between cooking coal smoke and the examined health outcomes. We conclude that exposure to heating coal smoke could have adverse effects on children's respiratory symptoms and illnesses in these four Chinese cities. PMID- 15118750 TI - Exposure to air pollutants in English homes. AB - BRE has conducted a national representative survey of air pollutants in 876 homes in England, designed to increase knowledge of baseline pollutant levels and factors associated with high concentrations. Homes were monitored for carbon monoxide (CO), nitrogen dioxide (NO(2)), formaldehyde and volatile organic compounds (VOCs). In the majority of the homes, concentrations of the measured pollutants were low. However, some homes have concentrations that would suggest a need for precautionary mitigation. Those factors that are most likely to lead to exposures of concern in homes are identified as gas cooking (for CO and NO(2)), the use of unflued appliances for heating (for CO and NO(2)), emissions from materials in new homes (for total VOC (TVOC) and formaldehyde), and painting and decorating, with a significant increase in risk suspected to exist where there is not a place to store materials away from the living space (for TVOC). It is noteworthy that seasonal effects on CO and NO(2) were largely due to indoor sources. This would need to be considered when interpreting time series studies of the effect of outdoor air pollution on health. It is also of some significance that the critical factors are related much more to sources than to ventilation: source control is therefore, as would be expected, the most appropriate approach to reducing the risk of hazardous exposure to air pollutants in homes. PMID- 15118751 TI - Differences in source emission rates of volatile organic compounds in inner-city residences of New York City and Los Angeles. AB - The Toxics Exposure Assessment Columbia-Harvard (TEACH) Project characterized personal, indoor, and outdoor concentrations of a suite of volatile organic compounds (VOCs) for high school students living in New York City (NYC) and Los Angeles (LA). This paper presents the analysis of VOC measurements collected indoors and outdoors for 46 students' homes in NYC and for 41 students' homes in LA across two seasons. Dual-sorbent thermal desorption tubes were used for the collection of 15 VOCs and C(18) 2,4-dinitrophenylhydrazine-coated cartridges were used for the collection of seven aldehydes. Air-exchange rates (AERs) were also measured using a perfluorocarbon tracer gas method. The AERs were lower in the winter in both cities, averaging 1 h(-1) in NYC and 1.4 h(-1) in LA, compared with 1.8 h(-1) in NYC in the summer and 2.5 h(-1) in LA in the fall. Higher AERs were generally associated with lower indoor-outdoor ratios with significant differences for the compounds with indoor sources, including chloroform, 1,4 dichlorobenzene, and formaldehyde. Using a mass-balance model to account for AER and other housing parameters, effective source emission rates (SER) were calculated for each compound. Based on I/O ratios and source emission rates, VOCs could be divided into: (1). indoor-source-influenced compounds, (2). those with contributions from both indoor and outdoor sources, and (3). those with mostly outdoor sources. Significant indoor sources were found for the following six compounds (mean emission rates presented): chloroform (0.11 mg/h), 1,4 dichlorobenzene (19 mg/h), formaldehyde (5 mg/h), acetaldehyde (2 mg/h), benzaldehyde (0.6 mg/h), and hexaldehyde (2 mg/h). Although chloroform had variable I/O ratios across seasons, SERs, which accounted for AER, were similar in both cities for both seasons (e.g., LA means 0.12 and 0.11 mg/h in winter and fall, respectively). Formaldehyde had substantially higher indoor emission rates in the summer in NYC compared to winter (3.8 vs. 1.6 mg/h) but lower in the fall in LA compared to winter (4.3 vs. 5.0 mg/h). Uncertainty analysis determined that source strength calculations were not sensitive to measurement error for a subset of homes in LA. PMID- 15118752 TI - Childhood asthma and indoor woodsmoke from cooking in Guatemala. AB - We estimated the prevalence and severity of asthma, and the association with cooking on open wood fires, as preparation for a large-scale randomized field trial on effects of indoor air pollution and child health. This is one of the first systematic studies of asthma and indoor wood-smoke pollution and to our knowledge the first asthma study in a purely indigeneous population in Latin America. The mothers of 1058 children aged 4-6 years were interviewed, using the standardized ISAAC (International Study of Asthma and Allergies in Childhood) procedures and questionnaire. The study population is a Mam-speaking (Maya), indigenous group living at relatively high altitude (2000 m) in Western Guatemalan Highlands. We found that asthma prevalence is low among indigenous children in Guatemala, compared to other populations in Latin America. Only 3.3% of the children reported wheezing symptoms in the last 12 months, and 72% wheezing symptoms ever. The majority of the current wheezers had at least one of the criteria for severe asthma. The prevalence of all the symptoms of asthma was higher in children from households that used open fires compared to improved stoves with chimneys. In a logistic regression model, use of open fire for cooking was a significant risk factor for a number of asthma symptoms, with odds ratios varying from 2.0 to 3.5. Among the different cooking technologies (1 improved stove with chimney, 2-mixture of gas and open fire, 3-open fire) trends of higher prevalence with more pollution was found for some of the symptoms. Hence use of open fire for cooking, may be an important risk factor for asthma symptoms and severity. PMID- 15118753 TI - Ambient, indoor and personal exposure relationships of volatile organic compounds in Mexico City Metropolitan Area. AB - Air pollution standards and control strategies are based on ambient measurements. For many outdoor air pollutants, individuals are closer to their sources (especially traffic) and there are important indoor sources influencing the relationship between ambient and personal exposures. This paper examines the relationship between volatile organic compounds (VOCs) measured at central site monitoring stations and personal exposures in the Mexico City Metropolitan Area. Over a 1-year period, personal exposures to 34 VOCs were measured for 90 volunteers from 30 families living close to one of five central monitoring stations. Simultaneous 24-h indoor, outdoor and central site measurements were also taken. Dual packed thermal desorption tubes and C(18) DNPH-coated cartridges were used for sampling VOCs and these were analyzed by GC/MS and HPLC, respectively. A factor analysis of the personal exposure data aided in grouping compounds by the most likely source type: vehicular (BTEX, styrene and 1,3 butadiene), secondary formed or photochemical (most aldehydes), building materials and consumer products (formaldehyde and benzaldehyde), cleaning solvents (tetrachloroethene and 1,1,1-trichloroethane), volatilization from water (chloroform and trichloroethene) and deodorizers (1,4-dichlorobenzene). Mean ambient, indoor and personal concentrations were 7/7/14 microg/m(3) for benzene, 1/3/3 for 1,3-butadiene, 6/20/20 for formaldehyde and 3/9/50 for 1,4 dichlorobenzene. Geometric mean (GM) ambient concentrations of trichloroethene and carbon tetrachloride were similar to GM personal exposures. While outdoor and indoor home GM concentrations for most vehicular related compounds (benzene, MTBE, xylenes and styrene) were comparable, the GM personal exposures were twice as high. Indoor concentrations of 1,3-butadiene, 1,1,1-trichloroethane, tetrachloroethane, chloroform, formaldehyde, valeraldehyde, propionaldehyde and n butyraldehyde were comparable to personal exposures. For certain compounds, such as chloroform, aldehydes, toluene, 1,3-butadiene and 1,4-dichlorobenzene, GM personal exposures were more than two times greater than GM ambient measurements. PMID- 15118754 TI - Effect of environmental interventions to reduce exposure to asthma triggers in homes of low-income children in Seattle. AB - The effectiveness of community health workers (CHWs) assisting families in reducing exposure to indoor asthma triggers has not been studied. In all, 274 low income asthmatic children were randomly assigned to high- or low-intensity groups. CHWs visited all homes to assess exposures, develop action plans and provide bedding encasements. The higher-intensity group also received cleaning equipment and five to nine visits over a year focusing on asthma trigger reduction. The asthma trigger composite score decreased from 1.56 to 1.19 (Delta= 0.37, 95% CI 0.13, 0.61) in the higher-intensity group and from 1.63 to 1.43 in the low-intensity group (Delta=-0.20, 95% CI 0.004, 0.4). The difference in this measure due to the intervention was significant at the P=0.096 level. The higher intensity group also showed improvement during the intervention year in measurements of condensation, roaches, moisture, cleaning behavior, dust weight, dust mite antigen, and total antigens above a cut point, effects not demonstrated in the low-intensity group. CHWs are effective in reducing asthma trigger exposure in low-income children. Further research is needed to determine the effectiveness of specific interventions and structural improvements on asthma trigger exposure and health. PMID- 15118755 TI - Indoor exposures and acute respiratory effects in two general population samples from a rural and an urban area in Italy. AB - A study of indoor air exposures and acute respiratory effects in adults was conducted in the Po Delta (rural) and Pisa (urban) areas of Italy. Indoor exposures were monitored for nitrogen dioxide (NO(2)) and particulate matter <2.5 microm (PM(2.5)) for 1 week during the winter or summer in a total of 421 houses (2/3 in Pisa). Information on house characteristics, subjects' daily activity pattern and presence of acute respiratory symptoms was collected by a standardized questionnaire. Peak expiratory flow (PEF) maneuvers were performed by adult subjects four times daily; maximum amplitude and diurnal variation were taken into account. Indices of NO(2) and PM(2.5) exposures were computed as the product of weekly mean pollutant concentration by the time of daily exposure. Mean levels of pollutants were significantly higher in winter than in summer, regardless of the area. The relationship between exposure indices and acute respiratory symptoms was investigated only in winter. In spite of a slightly lower indoor level in the urban than in the rural area in winter (NO(2): 15 vs. 22 ppb; PM(2.5): 67 vs. 76 microg/m(3)), prevalence rates of acute respiratory symptoms were significantly higher in the urban than in the rural area. Acute respiratory illnesses with fever were significantly associated with indices of NO(2) (odds ratio (OR)=1.66; 95% CI=1.08-2.57) and PM(2.5) exposures (OR=1.62; 95% CI=1.04-2.51), while bronchitic/asthmatic symptoms were associated only with PM(2.5) (OR=1.39; 95% CI=1.17-1.66). PEF variability was positively related only to PM(2.5) exposure index (OR=1.38; 95% CI=1.24-1.54, for maximum amplitude; OR=1.37; 95% CI=1.23-1.53, for diurnal variation). In conclusion, indoor pollution exposures were associated with the presence of acute respiratory symptoms and mild lung function impairment in a rural and an urban area of Northern-Central Italy. PMID- 15118756 TI - Gallbladder cancer treatment using adenovirus expressing the HGF/NK4 gene in a peritoneal implantation model. AB - Gallbladder cancer cells are stimulated by hepatocyte growth factor (HGF) in vitro and in vivo. We constructed an adenovirus vector, AdCMV.NK4, carrying the HGF antagonist HGF/NK4 (NK4) and evaluated whether or not this vector can suppress the peritoneal implantation of gallbladder cancer in a novel peritoneal injury mouse model. A human gallbladder cancer cell line (GB-d1) and human peritoneal mesothelial cells infected with the adenovirus vector produced a substantial level of NK4 protein. An invasion of GB-d1 cells was determined by a coculture with AdCMV.NK4-infected human mesothelial cells in vitro. Both the invasion and migration of GB-d1 cells were dramatically inhibited by this vector in a multiplicity of infection (MOI)-dependent manner. GB-d1 cells were intraperitoneally injected into the nude mice with peritoneal injury, followed by either AdCMV.NK4 or a control vector (AdCMV.LacZ). The incidence and the size of the metastatic tumor drastically decreased by AdCMV.NK4 (MOI 100: n=4, P<.0001). Real-time PCR analysis revealed a transient elevation of mouse HGF mRNA expression at the peritoneal injury sites. AdCMV.NK4 has been suggested to induce the inhibition of the implantation and growth of gallbladder cancer cells in vivo through its anti-HGF activity, and the use of NK4 gene transfer could be an effective modality for preventing peritoneal metastasis of gallbladder cancer. PMID- 15118757 TI - Inhibition of B16BL6 tumor progression by coadministration of recombinant angiostatin K1-3 and endostatin genes with cationic liposomes. AB - Transfection of the antiangiogenic angiostatin and endostatin genes was shown to be an alternative to high-dose administration of angiostatin or endostatin proteins for cancer therapy. We have systematically investigated whether coadministration of the mouse angiostatin kringle 1-3 gene (pFLAG-AngioK1/3) and the endostatin gene (pFLAG-Endo) complexed with cationic liposomes exhibits enhanced therapeutic efficacy. In vitro, the coexpressed mixture of angiostatin K1-3 and endostatin more effectively reduced angiogenesis in chorioallantoic membranes than either angiostatin K1-3 or endostatin alone. In vivo, subcutaneous co-administration of pFLAG-AngioK1/3 and pFLAG-Endo lipoplexes more effectively inhibited vascularization in Matrigel plugs implanted in mice than either one alone. Additionally, subcutaneous administration of these genes inhibited the growth and formation of pulmonary metastases of B16BL6 melanoma cells in mice. Compared to treatment with an empty vector, treatment with pFLAG-AngioK1/3 plus pFLAG-Endo inhibited 81% of tumor growth, while treatment with pFLAG-AngioK1/3 or pFLAG-Endo inhibited tumor growth 70 and 69%, respectively. Cotreatment with the two plasmids after primary tumor excision induced a 90% inhibition of pulmonary metastases versus 79% for pFLAG-AngioK1/3 or 80% for pFLAG-Endo individually. These results suggest that combined administration of angiostatin K1-3 and endostatin genes complexed with cationic liposomes may be an innovated antiangiogenic strategy for cancer therapy. PMID- 15118758 TI - Photochemically enhanced gene transfection increases the cytotoxicity of the herpes simplex virus thymidine kinase gene combined with ganciclovir. AB - Tumor targeting is an important issue in cancer gene therapy. We have developed a gene transfection method, based on light-inducible photochemical internalization (PCI) of a transgene, to improve gene delivery and expression selectively in illuminated areas, for example, in tumors. In the present work, we demonstrate that PCI improved the nonviral vector polyethylenimine (PEI)-mediated transfection of a therapeutic gene, the 'suicide' gene encoding herpes simplex virus thymidine kinase (HSVtk). In U87MG glioblastoma cells in vitro, the photochemical treatment stimulated expression of the HSVtk transgene, and, consequently, enhanced cell killing by the subsequent treatment with the prodrug ganciclovir (GCV). When relatively low doses of DNA (1 microg/ml) and the PEI vector (N/P 4) were used, HSVtk gene transfection followed by the GCV treatment did not have an effect on cell survival unless the photochemical treatment was performed, which potentiated the cytotoxicity to 90%. These findings indicate that photochemical transfection allows: (i) selective enhancement in gene expression and gene-mediated biological effects (cell killing by the Hsvtk/GCV approach) in response to illumination; (ii) the use of low, suboptimal for the nonviral transfection methods without PCI, doses of both DNA and the vector, which may be relevant and advantageous for therapeutic gene transfer in vivo. PMID- 15118759 TI - MIA (melanoma inhibitory activity) promoter mediated tissue-specific suicide gene therapy of malignant melanoma. AB - Suicide gene therapy of malignant melanoma essentially requires efficient gene transfer and highly selective therapeutic gene expression. To achieve this, recombinant adeno-associated virus (rAAV) particles were constructed containing the tissue-specific promoter of the human melanoma inhibitory activity (hMIA) gene combined with four copies of the enhancer element of the murine tyrosinase gene. Three melanoma and one cervix carcinoma cell line were infected with rAAV particles carrying a reporter gene under control of the enhancer/hMIA promoter in order to determine transcriptional activity and specificity of this system. Viral particles containing the enhancer/hMIA promoter mediated reporter gene activity only in melanoma cells, whereas infection with a cytomegalovirus (CMV)-based promoter construct induced unspecific gene expression. Correspondingly, transient transduction with viral particles bearing the HSVtk gene under the control of the enhancer/MIA promoter elements followed by treatment with ganciclovir (GCV) resulted in growth inhibition only in melanoma cells, whereas the CMV promoter based construct induced unspecific cytotoxicity. In vivo experiments in nude mice demonstrated that tumors originating from human melanoma cells disappeared after stable, but not transient transduction with vectors bearing the HSVtk gene under the control of the enhancer/hMIA promoter in response to GCV application. In face of higher transduction efficiency, these rAAV particles might therefore be a useful tool for suicide gene therapy of malignant melanoma. PMID- 15118760 TI - Antiproliferative activity of a triplex-forming oligonucleotide recognizing a Ki ras polypurine/polypyrimidine motif correlates with protein binding. AB - The Ki-ras gene is frequently mutated and/or overexpressed in human cancer. Since it is suspected to play a key role in the pathogenesis of many tumors, there is interest to search for strategies aiming at the specific inhibition of this oncogene. In this paper, we investigated the capacity of a 20 mer G-rich oligonucleotide (ODN20) conjugated to high molecular weight monomethoxy polyethylene glycol (MPEG) to inhibit the expression of the Ki-ras gene and the proliferation of pancreatic cancer cells. The conjugate, MPEG ODN20, was designed to form a triplex with a critical pur/pyr sequence located in the promoter of the Ki-ras gene. To make the conjugate resistant to endogenous and exogenous nucleases, five phosphorothioate linkages were introduced in its backbone. Confocal microscopy and FACS experiments showed that MPEG ODN20 had a higher capacity to penetrate the cell membranes and accumulate in the nucleus of Panc-1 cells than ODN20. Incubation of Panc-1 cells with MPEG ODN20 reduced specifically the levels of Ki-ras mRNA and RAS protein p21RAS. A single-dose administration of MPEG ODN20 was sufficient to inhibit cell proliferation by about 50% compared with control. By contrast, the antiproliferative activity of the unconjugated ODN20 analog was found to be not significant. Band-shift and footprinting experiments showed that MPEG ODN20 formed a weak triplex (Kd approximately 1.5 microM at 37 degrees C, 50 mM Tris-acetate, pH 7.4, 10 mM NaCl, 10 mM MgCl2, 5 mM spermidine) with the Ki-ras pyr/pur motif, suggesting that its bioactivity can hardly be mediated by a triplex-based mechanism. Here, we provide evidence that, in vitro, ODN20 and MPEG ODN20 competitively inhibit the binding to the Ki-ras pur/pyr motif of a nuclear protein, suggesting that the activity of MPEG ODN20 occurs with an aptameric mechanism. The biological implications of this study are discussed. PMID- 15118761 TI - DNA vaccines against the human papillomavirus type 16 E6 or E7 oncoproteins. AB - DNA vaccines expressing the E6 or E7 oncoproteins of human papilloma virus type 16 (HPV-16) in either their wild-type form or fused to sequences that affect intracellular trafficking were tested for induction of protective immunity against tumor cell challenge in two models based on BALB/c and C57Bl/6 mice. The DNA vaccines to E7 gave uniformly disappointing results, while the DNA vaccine that expressed E6 linked to a viral leader sequence protected BALB/c mice against tumor cell challenge given before or after vaccination. The efficacy of this vaccine could be enhanced by a DNA vector prime/viral vector boost regimen. In contrast, priming of mice with the DNA vaccines to E7 reduced the efficacy of a viral vector expressing the same antigen. PMID- 15118762 TI - Histone deacetylase inhibitors upregulate expression of the coxsackie adenovirus receptor (CAR) preferentially in bladder cancer cells. AB - Studies on bladder cancer cell lines have shown that low adenoviral (Ad) infectivity is associated with low-level coxsackie adenovirus receptor (CAR) expression. Recently, we and others demonstrated a tumor stage- and grade dependent downregulation of CAR expression in a large series of clinical bladder cancer specimens. Here, we demonstrate adenoviral gene transfer can be markedly enhanced in bladder cancer cells by upregulation of CAR through the use of certain differentiating agents, including the histone deacetylase inhibitors (HDACI) trichostatin A and sodium phenylbutyrate. CAR upregulation to supraphysiologic levels was demonstrated by quantitative rt-PCR, Western blotting, flow cytometry and adenoviral gene transfer. Normal urothelial cells and CAR-positive papilloma cells (RT4) failed to demonstrate upregulation under the same conditions. Upregulation was cell cycle dependent, associated with increased adenoviral gene transfer and persisted for at least 7 days after a single treatment. Such upregulation, however, appears to be tumor cell specific, as other CAR-negative cell lines failed to demonstrate enhanced adenoviral gene transfer with the same treatments. These results provide a rational basis for combining HDACI therapy with gene therapy as a method of augmenting activity in bladder cancer, but this strategy may not be universally applicable to other cell types. PMID- 15118763 TI - Downregulation of Bcl-2, FLIP or IAPs (XIAP and survivin) by siRNAs sensitizes resistant melanoma cells to Apo2L/TRAIL-induced apoptosis. AB - Melanoma cells are relatively resistant to Apo2L/TRAIL (TNF-related apoptosis inducing ligand). We postulated that resistance might result from higher expression of inhibitors of apoptosis including Bcl-2, FLIP (FLICE-like inhibitory protein) or IAPs such as XIAP (X-linked inhibitor of apoptosis) or survivin. Compared to scrambled or mismatch controls, targeting individual inhibitors with siRNA (si-Bcl-2, si-XIAP, si-FLIP or si-Surv), followed by Apo2L/TRAIL resulted in marked increase in apoptosis in melanoma cells. Compared to Bcl-2 or FLIP, siRNAs against XIAP and survivin were most potent in sensitizing melanoma cells. A similar substantial increase in apoptosis was seen in renal carcinoma cells (SKRC-45, Caki-2), following the inhibition of either XIAP or survivin by siRNAs. Apo2L/TRAIL treatment in IAP-targeted cells resulted in cleavage of Bid, activation of caspase-9 and cleavage of PARP (poly ADP-ribose polymerase). Thus, Apo2L/TRAIL resistance can be overcome by interfering with expression of inhibitors of apoptosis regulating both extrinsic (death receptor) or intrinsic (mitochondrial) pathways of apoptosis in melanoma cells. PMID- 15118764 TI - Camptothecin induced mitochondrial dysfunction leading to programmed cell death in unicellular hemoflagellate Leishmania donovani. AB - The parasites of the order kinetoplastidae including Leishmania spp. emerge from most ancient phylogenic branches of unicellular eukaryotic lineages. In their life cycle, topoisomerase I plays a significant role in carrying out vital cellular processes. Camptothecin (CPT), an inhibitor of DNA topoisomerase I, induces programmed cell death (PCD) both in the amastigotes and promastigotes form of L. donovani parasites. CPT-induced cellular dysfunction in L. donovani promastigotes is characterized by several cytoplasmic and nuclear features of apoptosis. CPT inhibits cellular respiration that results in mitochondrial hyperpolarization taking place by oligomycin-sensitive F0-F1 ATPase-like protein in leishmanial cells. During the early phase of activation, there is an increase in reactive oxygen species (ROS) inside cells, which causes subsequent elevation in the level of lipid peroxidation and decrease in reducing equivalents like GSH. Endogenous ROS formation and lipid peroxidation cause eventual loss of mitochondrial membrane potential. Furthermore, cytochrome c is released into the cytosol in a manner independent of involvement of CED3/CPP32 group of proteases and unlike mammalian cells it is insensitive to cyclosporin A. These events are followed by activation of both CED3/CPP32 and ICE group of proteases in PCD of Leishmania. Taken together, our study indicates that different biochemical events leading to apoptosis in leishmanial cells provide information that could be exploited to develop newer potential therapeutic targets. PMID- 15118765 TI - Early work on the function of Bcl-2, an interview with David Vaux. PMID- 15118766 TI - A novel mechanism for HIV1-mediated bystander CD4+ T-cell death: neighboring dying cells drive the capacity of HIV1 to kill noncycling primary CD4+ T cells. AB - CD4+ T-cell death is a crucial feature of AIDS pathogenesis, but the mechanisms involved remain unclear. Here, we present in vitro findings that identify a novel process of HIV1 mediated killing of bystander CD4+ T cells, which does not require productive infection of these cells but depends on the presence of neighboring dying cells. X4-tropic HIV1 strains, which use CD4 and CXCR4 as receptors for cell entry, caused death of unstimulated noncycling primary CD4+ T cells only if the viruses were produced by dying, productively infected T cells, but not by living, chronically infected T cells or by living HIV1-transfected HeLa cells. Inducing cell death in HIV1-transfected HeLa cells was sufficient to obtain viruses that caused CD4+ T-cell death. The addition of supernatants from dying control cells, including primary T cells, allowed viruses produced by living HIV1-transfected cells to cause CD4+ T-cell death. CD4+ T-cell killing required HIV1 fusion and/or entry into these cells, but neither HIV1 envelope mediated CD4 or CXCR4 signaling nor the presence of the HIV1 Nef protein in the viral particles. Supernatants from dying control cells contained CD95 ligand (CD95L), and antibody-mediated neutralization of CD95L prevented these supernatants from complementing HIV1 in inducing CD4+ T-cell death. Our in vitro findings suggest that the very extent of cell death induced in vivo during HIV1 infection by either virus cytopathic effects or immune activation may by itself provide an amplification loop in AIDS pathogenesis. More generally, they provide a paradigm for pathogen-mediated killing processes in which the extent of cell death occurring in the microenvironment might drive the capacity of the pathogen to induce further cell death. PMID- 15118767 TI - Walking to school: incidental physical activity in the daily occupations of Australian children. AB - Children's participation in physical activity is declining, and nowhere is this more apparent than in the incidental activity of walking to school. The aim of this pilot study was to examine the extent to which Australian children walked to and from primary school, and to survey parents to identify factors influencing this behaviour. Parents of 164 students in Grades 1 7 (mean age 9.1 +/- 2.02 years) from a primary school comprising 360 students responded to a questionnaire regarding psychosocial and environmental factors thought to influence the means by which their children went to and from school. Results indicated that parent perception of the importance of physical activity, parents' individual history of transport to school as well as distance from school were the most statistically significant factors determining children's involvement in walking to and from school. The results of this study highlight the attitudes and experiences of parents in determining the extent to which children are involved in non-motorized access to school. Also implicated are organizational policies about geographical school regions. While this study is limited to one school community, further study is recommended with others to better confirm findings by examining socioeconomic, geographic and policy variables. Occupational therapists are challenged to examine ways in which incidental physical activity can be increased in the lives of young children. PMID- 15118768 TI - Functional predictors of school participation by children with disabilities. AB - This study examined the functional requirements that significantly predicted participation of US elementary school children with a variety of disabling conditions (N = 266), in seven different school settings: Transportation, Transitions, Regular classroom, Special classroom, Mealtime, Bathroom, and Playground. Performance on a number of setting-relevant tasks was expected to be predictive of meaningful participation in each school environment. Stepwise multiple regression analyses were conducted to identify the order of importance of the relevant variables as well as the smaller set of functional tasks that best predicted children's participation in each setting. The results revealed that successful participation in the different elementary school settings was strongly associated with performance of both physical and cognitive/behavioural activities. Furthermore, each setting had a unique set of predictors, suggesting that some aspects of function are context-specific. Findings from this study may inform therapists about the most relevant areas of function that support social and physical participation of children with disabilities who are included in regular schools. PMID- 15118769 TI - The development of the School Function Assessment Chinese version for cross cultural use in Taiwan. AB - The primary purpose of this research project was to translate and adapt the School Function Assessment (SFA), a standardized criterion-referenced instrument that measures school-related functional skills, for its cross-cultural use in Taiwan. The project consisted of four study phases: translation, cultural adaptation, pilot testing, and field testing for standardization. A series of rigorous procedures including the method of translation and back-translation, team consensus for cultural adaptation, and Rasch modelling techniques were used to address various dimensions of cross-cultural equivalence and psychometric properties of the translated SFA. The protocols that were developed as well as technical issues that were addressed in this project provide useful guidelines for international occupational therapists who are interested in translating and adapting instruments for cross-cultural use. PMID- 15118770 TI - Changing practice patterns of school-based occupational therapists in Israel. AB - Following the enactment of the Special Education Law in Israel in 1988, school based occupational therapists (OTs) were expected to modify their practice patterns by, providing services within the classroom or other educational settings, increasing collaboration with other team members, and implementing indirect intervention models such as monitoring and consultation. The purpose of this study was to examine the intervention patterns of school-based OTs in Israel. The study sample included 77 school-based OTs. The survey was carried out through a questionnaire which included: (1) background information, (2) implementation and perceptions towards intervention models, and (3) team-work practices of the OTs. The findings indicated that most of the time (76.9%) OTs still implement the 'direct therapy' model, working outside the classroom and focusing on improving performance components. These findings imply that school based OTs in Israel are still in the process of changing their practice patterns, and are mostly applying the medical model. However, due to the relatively small sample, it is important to repeat this survey using a larger sample of OTs who work in different settings and with various age groups. PMID- 15118771 TI - Occupational therapists' reported experiences using weighted vests with children with specific developmental disorders. AB - The purpose of this study was to identify specific practice patterns of a convenience sample of paediatric occupational therapists and the behavioural changes that they observe when they use weighted vests with children with developmental disorders. Although the practice of weighted vests is accepted clinically, there is little discussion of their use nor is there empirical evidence of their efficacy in the literature. A convenience sample of 51 occupational therapists from different geographic areas of the United States participated in a telephone survey about how they used weighted vests with specific children. This study was a follow-up to a mail survey about paediatric occupational therapists' opinions and general practice patterns with weighted vests. Although the interviewees observed some different behavioural changes in children with various developmental disorders when these children used weighted vests, their practice patterns in using the vests were similar across disabilities. The most common behavioural changes noted were increased attention and staying on task. Participants' opinions and practice patterns related to weighted vests are discussed. This study is not generalizable as it utilized a self-selected group of therapists. However, the findings from the qualitative data can provide direction for future quantitative studies by providing important data about practice patterns that may serve as independent variables examining the effectiveness of weighted vests. PMID- 15118772 TI - School-based occupational therapy: an international perspective. PMID- 15118773 TI - The increasing threat of personal watercraft injuries. AB - A trend of increasing number and severity of injuries associated with use of personal watercraft (PWC) has been noted as the use and popularity of PWC also rises. The rate of injuries secondary to PWC use is greater than that from other water sports. Multiple etiologies of injury have been reported, including closed head trauma, spinal injuries, facial fractures, chest trauma, abdominal injuries, and drowning. These injuries may occur from collision with other objects, falls from the PWC, and encounters with the hydrostatic jet stream. The most significant contributing factors to injury seem to be carelessness and inattention. However, some studies show that substance abuse may also contribute. Recommendations to reduce morbidity and mortality include using U.S. Coast Guard approved personal flotation devices (PFD), limiting use of PWC to trained adults, and improving recognition of significant PWC injury by medical personnel. PMID- 15118774 TI - Appropriate use of antibiotics: focus on acute otitis media. PMID- 15118775 TI - Family environment factors and substance abuse severity in an HMO adolescent treatment population. AB - To examine how parental limit setting, family conflict, and perception of family experience influence severity of alcohol and drug problems, and important gender differences in these relationships, we interviewed consecutive intakes, aged 12 to 18 years, at 4 chemical dependency programs of a large group-model nonprofit health maintenance organization (HMO) (n=419). The Family Conflict, Limit Setting, and Positive Family Experience scales correlated with substance dependence (p<0.01, p<0.01, p<0.05, respectively). Depression also correlated with family conflict (p<0.01), absence of limit setting (p<0.01), poor family experience (p<0.01) and dependence symptoms (p<0.01). Number of substance-using friends correlated with number of dependence symptoms (p<0.01). Gender differences included the following: (1) girls scoring higher in family conflict (p=0.0002), negative perceptions of family experience (p<0.0017), and lower in absence of limit setting (p<0.0001); (2) how family environment predicted problem severity: absence of limit setting was significant for boys and girls but family conflict for boys only; (3) girls had more dependence symptoms (p=<0.0001), psychiatric diagnoses (e.g., depression (p<0.0003), anxiety (p<0.0002), conduct disorder (p=0.07)), and substance-abusing family members (53 % versus 39%; p=0.006). To conclude, family and peers influence severity of alcohol and drug problems in adolescents. PMID- 15118776 TI - Pediatric farm-related injuries: a series of 96 hospitalized patients. AB - Ninety-six children were admitted during a 9-year period to a pediatric level 1 trauma center for treatment of farm-related injuries. The age range was from 6 weeks to 17 years (median, 7.5 years; mean, 7.6 years; standard deviation, 4.4). Thirty-nine patients (40.6%) had an animal-related injury, including 36 children (37.5%) who had an injury associated with a horse. Amish children had an increased risk of horse-related injury when compared with non-Amish children (p=0.04; RR=2.09, 95% CI: 1.18SB203186>serotonin, cisapride, tropisetron>renzapride, 5-MeOT>5-CT). In transiently transfected HEK293 cells cisapride was a partial agonist compared to serotonin at 5-HT4(b), 5-HT4(g) and 5-HT4(i) receptors. In membranes from HEK293 cells stably expressing 5-HT4(g) (3,000 fmol/mg protein) or 5-HT4(i) (500 fmol/mg protein), serotonin and 5-MeOT were full agonists while cisapride was full agonist at 5-HT4(g) and partial agonist at 5-HT4(i), probably due to different receptor expression levels. At both 5-HT4(g) and 5-HT4(i), the behaviour of 5-HT4 receptor antagonists was dependent on receptor level. At high receptor levels, tropisetron and SB207710 and to a variable extent SB203186 and GR113808 displayed some partial agonist activity, whereas GR125487 and SB207266 reduced the AC activity below basal, indicating both receptors to be constitutively active. We conclude that the novel 5-HT4(i) receptor splice variant is pharmacologically indistinguishable from other 5-HT4 splice variants and that the 5-HT4(i) C terminal tail does not influence coupling to AC. PMID- 15118811 TI - Purification and characterization of 2,6-dihydroxybenzoate decarboxylase reversibly catalyzing nonoxidative decarboxylation. AB - A nonoxidative decarboxylase, 2,6-dihydroxybenzoate decarboxylase, was found in Agrobacterium tumefaciens IAM12048. The enzyme activity was induced specifically by 2,6-dihydroxybenzoate. The purified enzyme was a homotetramer of identical 38 kDa subunits. The purified decarboxylase catalyzed the nonoxidative decarboxylation of 2,6-dihydroxybenzoate and 2,3-dihydroxybenzoate without requiring any cofactors. In the presence of KHCO(3), the enzyme also catalyzed the regioselective carboxylation of 1,3-dihydroxybenzene into 2,6 dihydroxybenzoate at a molar conversion ratio of 30%. PMID- 15118809 TI - The effect of the alpha2-adrenoreceptor antagonist idazoxan against 6 hydroxydopamine-induced Parkinsonism in rats: multiple facets of action? AB - The alpha2-adrenoreceptor antagonist idazoxan counteracts catalepsy induced by neuroleptic agents and improves Parkinsonian signs in 1-methyl-4-phenyl 1,2,3,6,tetrahydropyridine-treated monkeys and in patients. The present study addressed the question of whether systemic administration of idazoxan (1.5 mg/kg i.p.) improves Parkinsonian symptoms in a rat model of permanent dopaminergic neurodegeneration. Dopaminergic degeneration was induced by injection of 6 hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB). Parkinsonian behaviour was assessed by catalepsy and open-field exploratory behaviour tests. Since dopaminergic and serotonergic mechanisms are thought to account for the anti-cataleptic/anti-Parkinsonian property of idazoxan, dopamine, 5 hydroxytryptamine (5-HT) and its metabolites in the regions of the basal ganglia and prefrontal cortex were analysed by HPLC. 6-OHDA lesions in the MFB produced catalepsy and hypoactivity in the open field and depleted dopamine and its metabolites in the basal ganglia and prefrontal cortex, but did not affect 5-HT. Treatment with idazoxan counteracted the observed Parkinsonian behaviour in 6 OHDA-lesioned rats and increased the spontaneous open-field activity in control rats. In both 6-OHDA and control animals, idazoxan increased DA level in the prefrontal cortex, but not in any other structures including the striatum. Idazoxan also increased the levels of 5-HT in the anterior striatum, prefrontal cortex and the ventral tegmental area of both 6-OHDA and control animals. These findings indicate that systemic administration of idazoxan counteracts 6-OHDA induced Parkinsonian symptoms in rats and that both dopaminergic and serotonergic mechanisms could contribute to its anti-Parkinsonian effect. PMID- 15118812 TI - Quality of life in ambulatory postmenopausal women: the impact of reduced bone mineral density and subclinical vertebral fractures. AB - Health-related quality of life (HRQOL) in postmenopausal women with osteoporosis has hitherto been mainly assessed in patients with clinically recognized vertebral fractures. Our study aimed to investigate the QOL perception in 361 asymptomatic ambulant postmenopausal women who came to our center for an osteoporosis screening program planned with their general practitioners. The Quality of Life Questionnaire of the European Foundation for Osteoporosis (QUALEFFO) was administered to all subjects. The participants underwent bone mineral density (BMD) measurements by DXA of either the lumbar spine and/or the femoral neck, as well as X-ray examination of the thoracolumbar spine to identify subclinical vertebral fractures. According to the WHO definition, where subjects are subdivided by BMD values into three groups (women with normal BMD, osteopenia, and osteoporosis), a significant difference was found only for the domains which explore general health perception (p<0.01 by ANOVA) and mental function (p<0.001 by ANOVA). When we segregated both osteopenic and osteoporotic women according to whether or not they had vertebral fractures, a significant difference was found only in osteoporotic patients for domains which explore physical function (p<0.001), social function (p<0.001), general health perception (p<0.02), and total QUALEFFO score (p<0.01). Stepwise multiple logistic regression analysis of the whole sample showed that both vertebral fractures and a low femoral BMD impairs QOL perception, while age did not exert a significant influence. ROC curves analysis demonstrated a low discriminating capacity of individual domains and total QUALEFFO score for both vertebral deformities and BMD categorization. Our results showed that QUALEFFO is not able to discriminate between patients with or without subclinical vertebral fractures. However, some aspects of QOL appear to be impaired in patients with subclinical vertebral fractures or reduced BMD. PMID- 15118813 TI - Risk factors for foot fracture among individuals aged 45 years and older. AB - A case-control study undertaken among members of five Northern California Kaiser Permanente medical centers sought to identify risk factors for foot fractures among persons aged 45 years and older. Foot fracture cases (n=920) and frequency matched controls (n=2366) were interviewed between October 1996 and May 2001 using a standardized questionnaire. Foot fractures occurred most often while walking or climbing stairs. While 60% of foot fractures resulted from falls, 20% were attributed to other causes, such as hitting the foot or tripping on sidewalks and curbs. Having a self-reported history of physician-diagnosed diabetes [adjusted odds ratio (OR)=1.45, 95% confidence interval (CI)=1.10-1.91] or cataracts (OR=1.40, 95% CI=1.07-1.83), having a self-reported foot problem (OR=1.38, 95% CI=1.06-1.78 for two or more foot problem versus no foot problems), having difficulty walking in minimum light (OR=1.86, 95% CI=1.14-3.05) and having had a prior fracture (OR=1.20, 95% CI=1.05-1.37) were associated with increased risk. Putative protective factors for osteoporotic fractures, such as menopausal hormone therapy use, thiazide or water pill use, high calcium intake, and high body mass index were not associated with foot fracture risk. These findings suggest that risk factors for foot fractures among older people differ in part from risk factors for other fracture sites generally considered to be osteoporotic, such as the hip, vertebrae, and forearm. PMID- 15118814 TI - Comparative study of knot performance and ease of manipulation of monofilament and braided sutures for arthroscopic applications. AB - Our aim was to identify alternative suture materials that might provide superior knot performance and equivalent ease of manipulation by means of an in vitro experimental study. Although used widely for arthroscopic shoulder stabilisation, absorbable poly( p-dioxanone) (PDS) monofilament sutures can lead to dehiscence and clinical failure due to knot slippage and/or loop elongation at low applied loads. With the objective of identifying alternative suture materials that might provide superior knot performance and equivalent ease of manipulation, an in vitro experiment was undertaken in which both Duncan and Snyder knotted loops were tied from four different suture materials using an arthroscopic knot pusher (Surgeon's 6th Finger) and a wooden practice box (Arthrex). Three monofilament sutures, made from polypropylene (PROLENE), poly(glycolidetrimethylenecarbonate co-dioxanone) (Biosyn) and PDS, and one braided polyester suture (SURGIDAC) were tested. Ease of manipulation was evaluated by measuring the time required to tie the knots, and the knot performance was assessed in terms of loop elongation and loop holding capacity by performing cyclic fatigue testing of the loops while they were immersed in saline on an INSTRON mechanical tester. The PROLENE and SURGIDAC sutures showed improvements in loop elongation and loop holding capacity over PDS, with the Snyder knots being consistently superior to Duncan knots. BIOSYN gave no improvement over PDS. Difficulties were encountered in advancing half-hitch throws of the braided SURGIDAC suture down the cannula, which resulted in longer knotting times. In conclusion, both the polypropylene and polyester sutures gave superior knot performance to PDS, but only the polypropylene gave equivalent ease of manipulation. As a result there may be clinical advantages in replacing PDS with monofilament polypropylene or braided polyester sutures for certain arthroscopic procedures. PMID- 15118815 TI - Influence of nitric oxide on microcirculation in pancreatic ischemia/reperfusion injury: an intravital microscopic study. AB - Recently, protective effects of nitric oxide donors in pancreatic ischemia/reperfusion (IRI) injury have been described. Their role in post ischemic microcirculation was previously not investigated. Ischemia reperfusion was induced in an isolated pancreatic tail segment in situ. Animals were randomized to four experimental groups (n=7 animals/group), the control group (CO) received saline as placebo. Treatment groups received either sodium nitroprusside (SN) 5 min before until 2 h after reperfusion, L-arginine (LA) 30 min before reperfusion until 2 h after reperfusion or sodium nitroprusside and L arginine (SNLA) together. After induction of ischemia (2 h) post-ischemic microcirculation was observed for 2 h by intravital-fluorescence microscopy. Functional-capillary density (FCD), leukocyte adherence in post-capillary venules (LAV) and histological damage were analysed. After reperfusion FCD decreased in all groups (P<0.05). FCD was significantly restored in all groups with administration of nitric oxide donors after reperfusion (P<0.05) as compared to CO without significant difference between the individual nitric oxide donor groups. Leukocyte adherence was significantly increased 1 h and 2 h after reperfusion (P<0.001) as compared to baseline, which was lower in all nitric oxide donor groups. Histological damage in the pancreatic tail-segment was significantly reduced in nitric oxide donor groups (P<0.01). Administration of nitric oxide donors might be useful in ischemia-reperfusion injury of the pancreas by its protective effect on microcirculation and inflammatory reaction. PMID- 15118816 TI - Use of percutaneous tracheostomy in intensive care units in Spain. Results of a national survey. AB - OBJECTIVES: To assess the use of percutaneous tracheostomy in Intensive Care Units (ICU) in Spain, its practice, and current opinions on the technique. DESIGN AND SETTING: An e-mail or post survey was sent to 239 Spanish ICU directors. Pediatric ICUs and coronary units were excluded. MEASUREMENTS AND MAIN RESULTS: One hundred ICUs (41.8%) replied. The 44% ( n=44) of the ICUs that answered belonged to university hospitals and 53% ( n=53) had postgraduate teaching. Eighty-two percent ( n=82) used percutaneous tracheostomy. Griggs' Guide Wire Dilating Forceps and Ciaglia Blue Rhino were the most frequent techniques employed. In 30.5% of ICUs ( n=25) endoscopic guidance was used, in 15.7% ( n= 13) it was routine. In 24.4% ( n=20) some kind of long-term follow-up was carried out, but only in 12.2% ( n=10) was follow-up done routinely. In 58.5% of ICUs ( n=48) in which percutaneous tracheostomy is performed is this technique considered safer than surgical tracheostomy and in 86.4% ( n=70) percutaneous tracheostomy is the first choice for tracheostomy in the critically ill patient. CONCLUSIONS: Percutaneous tracheostomy is a well-established technique in ICUs in Spain, and is considered the technique of choice for tracheostomy in critically ill patients. It is mainly performed without endoscopic guidance and follow-up is not usually carried out. PMID- 15118817 TI - Safe paediatric intensive care. Part 2: workplace organisation, critical incident monitoring and guidelines. AB - In order to optimise safety within the paediatric intensive care unit (PICU), it is essential to optimise organisation, identify problem areas and implement standards and guidelines for safe practice (with appropriate monitoring). Organisational issues have a major impact on safety: the introduction and recently-centralisation of paediatric intensive care, the appointment of dedicated paediatric intensivists, nursing staffing, handovers, rounds, the number of work hours and night shifts with the associated problems of disturbed circadian rhythms. The technique of voluntary, anonymous, non-punitive critical incident reporting has the potential to identify incidents and latent errors before they become self-evident through a major incident. This systems approach focuses on organisational and communication problems. Standards and guidelines may help in weighing up the benefits and risks of invasive procedures, and interventional studies have shown that implementation of standards and guidelines can improve outcome. Mortality prediction models enable us to monitor quality of care and, thus, to investigate the best ways of organising intensive care and monitoring the effects of changes in practice. PMID- 15118818 TI - Does red blood cell transfusion change the near infra red photoplethysmography signal in infants? AB - OBJECTIVE: Transfusion practices for neonates are controversial and based on limited scientific information. We examined the use of the plethysmographic volume pulse to assess anemia. DESIGN: We used near-infrared photoplethysmography (NIRP) to investigate signal strength parameters as area under the curve (AUC) and the first derivate of the amplitude (flux) in anemic infants before and after elective transfusion. SETTING: Tertiary intensive care unit. PATIENTS: Thirty four neonates on day 25+/-15 of life with a pretransfusion hemoglobin level of 8+/-0.9 g/dl. Four infants were excluded for incomplete data. INTERVENTIONS: Transfusion of 10 ml packed red cells per kilogram body weight MEASUREMENTS AND RESULTS: The volume pulse signal was obtained with a sensor attached to the sole of the foot. After transfusion we found a significant decrease in number of bradycardia episodes per hour (0.35 vs. 0.16) and episodes of pulse oximeter desaturation less than 85% per hour (1.8 vs. 1.3) and a significant increase in daily weight gain (12+/-7 vs. 20+/-18 g), but neither of the microcirculatory parameters changed significantly (AUC 102+/-38 vs. 101+/-44; flux 435+/-160 vs. 405+/-120). CONCLUSIONS: Even though infants were transfused at very low levels of hemoglobin with significant clinical effects, microcirculation assessed by NIRP seemed not be affected. Infrared photoplethysmographs present flow not only in the nutritive capillaries but mainly in the subpapillary plexus at a greater depth. This thermoregulatory flow seems not be affected by blood transfusions of anemic infants. PMID- 15118819 TI - [Introduction to the topic: Pain therapy. New knowledge alters orthopedic management strategies]. PMID- 15118820 TI - [Primary neurogenic and myogenic disorders of posture]. AB - Disturbance of posture may occur in a variety of neurological disorders and occasionally is the presenting or even the only sign. In the majority of cases, the head or the trunk or both are bent forward (bent spine syndrome, dropped head syndrome). A feature of these primary neurogenic or myogenic postural disturbances that is in contrast to antalgic contraction or ankylosis is that they are not fixed, but the trunk or head are easily erected by the examiner and show a characteristic sagging. Neuromuscular disorders are a frequent cause. They may be confined to the paraspinal muscles. Axial computed tomography of the spine, electromyography of the involved muscles, and muscle biopsy help to make the diagnosis. However, also central movement disorders may lead to a sagging of the head or trunk or of both due to a lessened tone of the head and trunk extensors. This is frequently seen in the various parkinsonian syndromes which may, however, occur in association with a focal myopathy of the paraspinal muscles. Occasionally, sagging of the trunk is seen as a side effect of neuropharmacologic medication. Sagging of the trunk or head should be differentiated from a pathologically increased innervation of the ventral muscles in dystonic movement disorders such as antecollis or camptocormia. Pathologic reclination of the head or trunk or both is a rare disturbance of posture. It may occur in dystonia (retrocollis) or, occasionally, as a consequence of musculotendinous contractures secondary to certain neuromuscular disorders such as the rigid spine syndrome. PMID- 15118821 TI - [Metal/metal--a new (old) hip bearing system in clinical evaluation. Prospective 7-year follow-up study]. AB - The problems of wear debris of bearing systems containing polyethylene used in hip arthroplasty have led to an increased trend to hard on hard bearing surfaces. Based on our own good experiences with the cementless Zweymuller-Alloclassic system, we implanted the same system with metal-on-metal bearing surfaces (Metasul) in 100 cases between October 1993 and November 1994. In order to record potential side effects, specific questioning and a clinical and radiological survey were carried out before surgery and in regular intervals thereafter. Up to date almost all of the patients have had good and excellent results with an average of more than 90 points in the Harris hip score (47.8 before surgery). There were no cases of revision surgery as a result of aseptic loosening. Two revisions were performed for other reasons (change of the insert, deep infection). No disadvantageous phenomena related to Metasul in the surrounding tissue were found on macroscopic and histologic investigation. The radiological examination did not show any specific signs pertaining to the use of a metal-on metal bearing system. We also have no grounds to suppose that there are systemic reactions due to chromium or cobalt ions. Up to now, the new metal-on-metal bearing system in connection with a proven hip arthroplasty system has lived up to clinical expectations. With regard to less wear debris, it is justified to implant it in younger patients as well. Other open questions, in particular the possibly improved long-term results, have to be answered by continuation of the study. PMID- 15118822 TI - High-resolution genetic mapping of Xa27(t), a new bacterial blight resistance gene in rice, Oryza sativa L. AB - Bacterial blight of rice, caused by Xanthomonas oryzae pv. oryzae ( Xoo) (Ishyama) Dye, is one of the serious diseases prevalent throughout Asia. In a previous study, a resistance ( R) locus was transferred from the tetraploid wild rice Oryza minuta to the cultivated rice species, Oryza sativa L. Here, we report the fine genetic mapping of the R locus, tentatively designated as Xa27(t). We performed disease evaluation with an Xa27(t) near-isogenic line, IRBB27, testing 35 Xoo strains collected from 11 countries. The Xa27(t) locus conferred a high level of resistance to 27 strains and moderate resistance to three strains. Resistance of the Xa27(t) gene was developmentally regulated in IRBB27 and showed semi-dominant or a dosage effect in the cv. CO39 genetic background. As a prelude to cloning Xa27(t), a molecular mapping strategy was employed with a large mapping population consisting of 3,875 gametes. Three molecular markers, M336, M1081, and M1059, closely linked to Xa27(t), were identified to facilitate the mapping of Xa27(t) to the long arm of chromosome 6. The Xa27(t) locus was confirmed by chromosome landing of M1081 and M1095 markers on the rice genome. Markers derived from the genomic sequence of O. sativa cv. Nipponbare were used to further saturate the Xa27(t) genomic region. Xa27(t) was finally located within a genetic interval of 0.052 cM, flanked by markers M964 and M1197, and co segregated with markers M631, M1230, and M449. PMID- 15118823 TI - [Ultrastructural changes in the aging bladder]. AB - Recent prospective, comparative ultrastructural/urodynamic studies on geriatric voiding dysfunction, including its natural evolution, have revealed that abnormalities of detrusor behavior have specific correlates in detrusor microstructure represented by distinctive electron-microscopic patterns. Each electron-microscopic pattern comprises a constellation of features in two or all three tissue compartments of the detrusor (smooth muscle, interstitium, intrinsic nerves). The pattern characteristic of each dysfunction is the same in detrusors of men and women. The electron-microscopic patterns are additive when multiple abnormalities coexist. All distinctive patterns of various clinical voiding dysfunctions can readily be defined qualitatively in endoscopic biopsies of the detrusor. These observations, including those on material studied for the natural evolution of geriatric voiding dysfunction, have led to the proposal of a clinical/structural definition of the normal aging detrusor and the development of an algorithm for pathologic diagnosis of geriatric voiding dysfunction. PMID- 15118824 TI - [Changes in brain structure in bipolar affective disorders]. AB - The neurobiological basis of bipolar affective disorders is unknown. However, neuroanatomic circuits of mood regulation have been hypothesized. Neuroimaging revealed volumetric changes of specific brain structures in these circuits. The most prominent abnormality is enlargement of the amygdala. In addition there might be structural changes in the frontal lobe, cerebellum, and pituitary. The findings in bipolar disorder differ from those in unipolar depression and schizophrenia. For further identification of the neurobiological basis of bipolar disorders, structural neuroimaging combined with functional neuroimaging such as magnetic resonance spectroscopy, neuroendocrinological studies, and genetical analyses are required to subgroup patients with bipolar disorder by diagnostic, prognostic, and therapeutic criteria. PMID- 15118825 TI - [Factors influencing secondary preventive blood pressure control in hypertensive stroke patients]. AB - BACKGROUND: More than half of patients with documented hypertension surviving cerebrovascular events do not have their blood pressure (BP) controlled, despite the use of antihypertensive medication. Data on the possible reasons for poor BP control in stroke survivors are limited. METHODS: We evaluated prospectively parameters influencing the effectiveness of BP control in hypertensive stroke survivors who were followed up. RESULTS: At 3 months after stroke BP was effectively controlled in only 38.8% of patients. Diabetes mellitus had a negative influence on the effectiveness of antihypertensive medication by reducing significantly the chance of achieving a significant BP reduction, while patients suffering from coronary artery disease had an increased chance of getting their BP under control within 3 months after stroke onset. Diuretic medication was found to be an independent positive predictor for effective antihypertensive control. Combined therapy including diuretics was significantly more effective than antihypertensive monotherapy. CONCLUSIONS: Arterial pressure control in stroke survivors is poor. Risk factor profile, manifest heart disease, and the chosen antihypertensive medication are factors of prognostic relevance for effective BP control. PMID- 15118826 TI - [Correlation between clinical apparatus-based differential diagnosis and neuropathological diagnosis in patients with AIDS]. AB - BACKGROUND: During the course of HIV infection, the majority of patients develop opportunistic cerebral neuro-manifestations. If conventional diagnostic tools are not sufficient, a stereotactic biopsy is often necessary. PATIENTS AND METHODS: In order to evaluate the correctness of the clinical diagnosis of cerebral neuro manifestations in HIV-infected patients, we compared the results of cerebral biopsy or autopsy with the previous clinical diagnosis. A total of 19 biopsies and 49 autopsies could be analyzed. RESULTS: Except for HIV-associated encephalopathy, we detected a very high conformity between the clinical and the neuropathological diagnoses. We obtained the best sensitivity for progressive multifocal leukoencephalopathy (PML), whereas for cerebral toxoplasmosis the worst sensitivity and specificity was identified. CONCLUSION: We conclude that the diagnosis of PML can be made on clinical grounds alone, whereas the diagnosis of cerebral toxoplasmosis and lymphoma often requires a biopsy, which should be performed early. PMID- 15118827 TI - [Normalization of the Brief Repeatable Battery of Neuropsychological tests (BRB N) for German-speaking regions. Application in relapsing-remitting and secondary progressive multiple sclerosis patients]. AB - The Brief Repeatable Battery of Neuropsychological Tests (BRB-N) is a neuropsychological screening battery, often applied in multiple sclerosis (MS) patients. It is almost exclusively administered in trials and not in the daily practice routine because of the lack of normative values. Using a stepwise linear regression analysis, the dependence of test results on age, gender, and education of 241 healthy control subjects was investigated. Z-values of -1.68 or less were considered pathological. Based on the normative values, the proportions of cognitively impaired patients with relapsing-remitting MS (RRMS, n=43) and secondary progressive MS patients (SPMS, n=60) were calculated. The regression model explained 2.7-25.0% of the variance of test performances. Cognitive impairment occurred in 38% and in 47% of the RRMS and the SPMS groups, respectively. In both groups attention and concentration impairment was prominent, while in the SPMS group memory was also frequently affected. The proportion of cognitively impaired MS patients reflected the figures that could be found in the literature. PMID- 15118828 TI - [Differences between anteroposterior and posterior spondylodesis in clinical scores]. AB - The present study on spondylitis patients was carried out to compare prospectively results of clinical scores of patients treated with anteroposterior or anterior spondylodesis. A total of 22 patients with spondylitis were enrolled in the randomized trial to undergo an operation with anteroposterior or posterior spondylodesis. For quality of life adjustment, the SF-36 health survey was applied. Pain and disability were measured using the Oswestry low-back pain disability questionnaire and visual analogue scales (VAS). Twelve patients were assigned to undergo anteroposterior spondylodesis and ten patients anterior spondylodesis. Postoperative improvement of spinal function, general health, well being, back pain, and mobility could be observed in both groups of patients. Better results of patients treated with anterior spondylodesis compared with anteroposterior spondylodesis were seen. No statistical difference was found between the two groups at any time. The less invasive nature of anterior spondylodesis could be responsible for that result.Anterior spondylodesis should used if possible and deemed adequate because it causes less discomfort. PMID- 15118829 TI - [Hepatitis as a travel disease]. AB - Every year over 50 million people travel from industrialized countries to areas with high prevalence of oral-fecal and sexually transmissible forms of viral hepatitis. The risk of infection with hepatitis A is associated with the standard of living, the length of stay, and the area of destination. Acute hepatitis E is predominantly transmitted in India and other Asian countries. The main risk factors for the acquisition of hepatitis B are sexual promiscuity and unprotected sexual intercourse. This report provides detailed information on the risk of hepatitis in travelers, available vaccination schedules, clinical and laboratory diagnostic features, and necessary therapeutic aspects in cases of ongoing acute viral hepatitis. PMID- 15118830 TI - Endoscopic third ventriculostomy in infants of less than 1 year of age: which factors influence the outcome? AB - OBJECTIVES: Endoscopic third ventriculostomy (ETV) is a successful method of treatment for obstructive hydrocephalus. In infants, however, it is reported to have a higher failure rate. On the basis of our own data and a meta-analysis of the literature, we try to define factors prognosticating potential failure in infants aged less than 1 year. METHODS: Data were collected retrospectively. Between October 1994 and October 2002, 20 ETVs were performed in 16 patients younger than 1 year. Ages ranged from 8 to 311 days (median 103). Etiology was aqueductal stenosis in all 16 patients (idiopathic in 7, posthemorrhagic in 3, postmeningitic in 3, and related to CNS or vascular malformation in 3). ETV failure was defined as subsequent need for shunt implantation. For non-shunted patients, follow up was 16-52 months (median 25). RESULTS: ETV was successful in 5 patients and eventually failed in 11. There was no mortality or permanent morbidity following ETV. In the successful cases, etiology was idiopathic aqueductal stenosis in 4 and postmeningitic aqueductal stenosis in 1; the median age was 206 days (range 82-311). In the 11 unsuccessful patients, it was idiopathic aqueductal stenosis in 3, posthemorrhagic in 3, postmeningitic in 2 and CNS/vascular malformation in 3 cases; median age was 94 days (range 8-299). Median time interval between (last) ETV and shunt was 38 days (range 2-70). The difference in median age between the success group and the failure group roughly corresponded to data gained from a meta-analysis of the literature. Four patients underwent a second ETV. In intraoperative ventriculoscopy, the stoma was closed or there were new membranes below the floor of the third ventricle and a second ETV was performed. But finally, all re-ETVs failed and the patients needed a shunt. CONCLUSION: Factors indicating potential failure of ETV were very young age and etiology other than idiopathic aqueductal stenosis. Probability of success seems to increase during the first 2 or 3 months of life. Ventriculoscopy with the option of a second ETV should be regularly performed after failure of ETV. PMID- 15118831 TI - Near-infrared fluorescence imaging of HER-2 protein over-expression in tumour cells. AB - The aim of this study was to evaluate in vitro and in vivo imaging of HER-2-over expressing tumours using near-infrared optical imaging. A fluorochrome probe was designed by coupling Cy5.5 to anti-HER-2 antibodies. Cells over-expressing (SK-BR 3 cells) or normally expressing (PE/CA-PJ34 cells) the HER-2 protein were incubated with the probe. After removing unbound probe molecules, fluorescence intensities were determined (a.u.: arbitrary units). Cells were additionally investigated using FACS and laser scanning microscopy. The probe was also injected intravenously into tumours bearing SK-BR-3 ( n=3) or PE/CA-PJ34 ( n=3). Whole-body fluorescence images were generated and analysed. The incubation of SK BR-3 cells with the probe led to higher fluorescence intensities [2,133 (+/-143) a.u.] compared to controls [975 (+/-95) a.u.]. The results from FACS and immunocytochemical analysis were in agreement with these findings. A distinct dependency between the fluorescence intensity and the cell number used in the incubations was detected. In vivo, the relative fluorescence intensities in SK-BR 3 tumours were higher than in PE/CA-PJ34 tumours at 16-24 h after probe application. HER-2-over-expressing tumours were depictable in their original size. Labelling of HER-2 with Cy5.5 is suitable for in vitro and in vivo detection of HER-2-over-expressing tumour cells. PMID- 15118832 TI - Characterization of a multifunctional methyltransferase from the orchid Vanilla planifolia. AB - The final enzymatic step in the synthesis of the flavor compound vanillin (4 hydroxy-3-methoxybenzaldehyde) is believed to be methylation of 3,4 dihydroxybenzaldehyde. We have isolated and functionally characterized a cDNA that encodes a multifunctional methyltransferase from Vanilla planifolia tissue cultures that can catalyze the conversion of 3,4-dihydroxybenzaldehyde to vanillin, although 3,4-dihydroxybenzaldehyde is not the preferred substrate. The higher catalytic efficiency of the purified recombinant enzyme with the substrates caffeoyl aldehyde and 5-OH-coniferaldehyde, and its tissue distribution, suggest this methyltransferase may primarily function in lignin biosynthesis. However, since the enzyme characterized here does have 3,4 dihydroxybenzaldehyde-O-methyltransferase activity, it may be useful in engineering strategies for the synthesis of natural vanillin from alternate sources. PMID- 15118833 TI - Induced mutations in cassava using somatic embryos and the identification of mutant plants with altered starch yield and composition. AB - A cyclic somatic embryogenic system was used to induce mutations in cassava variety PRC 60a in vitro. Globular-stage somatic embryos were selected as suitable experimental materials, and 50 Gy of gamma-rays was determined to be the optimal dose for inducing mutations. During subsequent field trials, more than 50% of the regenerated mutant lines varied morphologically from wild-type plants. Consequently, we used this approach to induce genetic variability for obtaining novel cassava cultivars. Among the different mutant lines obtained, lines S14 and S15 showed large morphological variations. In 10-month-old S14 and S15 mutant lines, storage root yield was reduced 17-fold and 60-fold, respectively, compared to wild-type plants, while the storage roots of S15 mutant plants also exhibited an almost 50% decrease in starch content and a significant reduction (30%) in amylose content. These two features were observed throughout the different developmental stages of the storage roots in S15 plants. PMID- 15118834 TI - Adventitious root formation in Arabidopsis thaliana thin cell layers. AB - This paper describes, for the first time, de novo adventitious root formation from thin cell layers (TCLs) of Arabidopsis thaliana. The objective of the study was to determine the optimal hormonal and light conditions and the optimal exogenous Ca2+ concentration for obtaining adventitious rooting (AR) from A. thaliana TCLs and to identify the tissue(s) involved in the process. The results show that maximum AR was obtained with a single-phase method in the presence of 10 microM indole-3-butyric acid and 0.1 microM kinetin under continuous darkness for 30 days and with 0.6 mM exogenous CaCl2. The endodermis was the only tissue involved in root meristemoid formation. The role of Ca2+ in AR and the importance of using Arabidopsis TCLs in studies on the genetic/biochemical control of AR are discussed. PMID- 15118835 TI - An analysis of the phylogenetic distribution of the pea pathogenicity genes of Nectria haematococca MPVI supports the hypothesis of their origin by horizontal transfer and uncovers a potentially new pathogen of garden pea: Neocosmospora boniensis. AB - The filamentous fungus Nectria haematococca mating population VI (MPVI) contains a cluster of genes required to cause disease on pea. This cluster of pea pathogenicity genes (the PEP cluster) is located on a supernumerary chromosome that is dispensable for normal growth in culture. The genes in the PEP cluster have a different G+C content and codon usage compared with the genes located on the other chromosomes and a non-homogeneous distribution within the species. These features suggest that the PEP cluster may have been acquired by N. haematococca MPVI through horizontal gene transfer (HGT). In this work, we show that homologues of the PEP genes are present in another pea pathogen, Fusarium oxysporum f. sp. pisi, but are not common among fungi that are phylogenetically closely related to N. haematococca MPVI. This phylogenetic discontinuity supports the hypothesis that the PEP cluster originated by HGT. Our analysis has also determined that homologues for all the PEP genes are present in Neocosmospora boniensis. A molecular characterization of the PEP homologues in this fungus shows that they are organized as a cluster, which has a different physical organization from the PEP cluster in N. haematococca. In addition, although no reports have been found to show that N. boniensis is a naturally occurring pea pathogen, we show here that this species is able to cause disease on pea. PMID- 15118836 TI - NY-ESO-1 expression and its serum immunoreactivity in esophageal cancer. AB - PURPOSE: NY-ESO-1, a member of the cancer/testis antigen (CTA) family, elicits humoral and cellular immune responses in patients with advanced cancer. Unresectable or metastatic esophageal carcinoma patients do not benefit from the present multimodality treatment regimens in terms of survival. The objectives of this study were to analyze the antibody response to NY-ESO-1 antigen in patients with esophageal cancer and to determine the potential of NY-ESO-1 for use in tumor-specific immunotherapy. METHODS: Serum from 69 patients with esophageal cancer was investigated for antibody production against NY-ESO-1 by Western blot analysis. Also analyzed by immunohistochemistry were 56 tissue samples from these patients for NY-ESO-1 protein expression. RESULTS: NY-ESO-1 protein expression was found in 18 of 56 (32%) esophageal carcinomas. Serum immunoreactivity specific for NY-ESO-1 was found in 9 patients (13%) of whom 8 were in the advanced stage (stages III and IV). There was no relationship between clinicopathologic features and serum immunoreactivity for NY-ESO-1. NY-ESO-1 protein expression was detected in three of five antibody-positive patients whose tissue was available for analysis. Survival analysis showed no significant difference between antibody-positive and antibody-negative patient groups. CONCLUSIONS: A humoral immune response to NY-ESO-1 antigen was established in patients with advanced esophageal cancer. NY-ESO-1 is a good candidate for vaccine-based immunotherapy for advanced esophageal carcinoma. PMID- 15118837 TI - In vivo imaging of hepatobiliary transport function mediated by multidrug resistance associated protein and P-glycoprotein. AB - Multidrug resistance associated proteins (MRPs) and P-glycoprotein (P-gp) are involved in hepatobiliary transport of various compounds. Our aim was (1) to define transporter specificity of the cholescintigraphic agents 99mTc-HIDA and 99mTc-MIBI, which are used clinically for myocardial perfusion measurements; and (2) to deduce MRP and P-gp functions in vivo from hepatic 99mTc kinetics. Accumulation of radioactivity was measured in the human tumor cell lines GLC4, GLC4/ADR150x (MRP1-overexpressing/P-gp-negative) and GLC4/P-gp (P-gp overexpressing). Bile secretion was quantified in untreated and in glutathione depleted control and MRP2-deficient (GY/TR-) rats. Hepatobiliary transport was measured using a gamma camera in both types of rats. 99mTc-HIDA accumulated 5.8 fold less in GLC4/ADR150x calls than in GLC4 or GLC4/P-gp cells. In GLC4/ADR150x, the cellular 99mTc-HIDA content was increased 3.4-fold by the MRP1,2 inhibitor MK571 (50 microM), while MK571 had no measurable effect in GLC4 and GLC4/P-gp cells. 99mTc-MIBI accumulated less in GLC4/P-gp and GLC4/ADR150x cells than in GLC4 cells. Bile secretion of 99mTc-HIDA was impaired in GY/TR- compared to control rats and not affected by glutathione depletion in GY/TR- rats. Hepatic secretion of 99mTc-HIDA was slower in GY/TR- (t1/2 40 min) than in control rats (t1/2 7 min). Bile secretion of 99mTc-MIBI was similar in both rat strains and impaired by glutathione depletion in control rats only, indicating compensatory activity of additional transporter(s) in GY/TR- rats. 99mTc-HIDA is transported only by MRP1,2 only, while 99mTc-MIBI is transported by P-gp and MRP1,2. The results indicate that hepatic P-gp and MRP1,2 function can be assessed in vivo by sequential use of both radiopharmaceuticals. PMID- 15118838 TI - Three steps to cancer: how phosphorylation of tubulin, tubulin tyrosine ligase and P-glycoprotein may generate and sustain cancer. AB - Transformed cells progress to cancer because they are not eliminated by apoptosis. In this brief minireview I propose, based on published data, that the cell possesses a 'last check point' (LCP) apoptotic step in the form of assembly of nitrotyrosinated alpha-tubulin onto microtubules. This leads to microtubule dysfunction and ultimately apoptosis. I also propose that cells that escape this LCP apoptotic step develop into cancer. Phosphorylation of tubulin tyrosine ligase (TTL) is postulated to cause escape from LCP apoptosis. Phosphorylation also ensures that cancer cells survive a hostile milieu (e.g. chemotherapy). PMID- 15118839 TI - Numeric and symbolic knowledge representation of cerebral cortex anatomy: methods and preliminary results. AB - The human cerebral cortex anatomy describes the brain organization at the scale of gyri and sulci. It is used as landmarks for neurosurgery as well as localization support for functional data analysis or inter-subject data comparison. Existing models of the cortex anatomy either rely on image labeling but fail to represent variability and structural properties or rely on a conceptual model but miss the inner 3D nature and relations of anatomical structures. This study was therefore conducted to propose a model of sulco-gyral anatomy for the healthy human brain. We hypothesized that both numeric knowledge (i.e., image-based) and symbolic knowledge (i.e., concept-based) have to be represented and coordinated. In addition, the representation of this knowledge should be application-independent in order to be usable in various contexts. Therefore, we devised a symbolic model describing specialization, composition and spatial organization of cortical anatomical structures. We also collected numeric knowledge such as 3D models of shape and shape variation about cortical anatomical structures. For each numeric piece of knowledge, a companion file describes the concept it refers to and the nature of the relationship. Demonstration software performs a mapping between the numeric and the symbolic aspects for browsing the knowledge base. PMID- 15118840 TI - Acetabular reconstruction in developmental hip dysplasia using reinforcement ring with a hook. AB - We followed prospectively 27 patients with severe acetabular bone-stock deficiencies due to developmental dysplasia of the hip. Mean preoperative patient age was 56 (34-78) years, mean Harris hip score was 31 (16-66) points and pre operative mean acetabular angle of Sharp was 47 degrees (34 degrees - 61 degrees). Operative acetabular reconstruction was performed in 28 hips using reinforcement ring with a hook placed in the true acetabulum and autologous bone grafting. A 2-year clinical and radiological follow-up was available in 27 hips. Mean Harris hip score improved to 91 (70-100) points. There were no clinical or radiological signs of aseptic loosening in 25 hips, and the bone graft was radiologically incorporated in all hips. In 24 hips, a radiostereometric analysis showed low rates of non-progressive translation and rotation in 21 reinforcement rings. Three rings showed progressive translation and/or rotation at the 2-year follow-up and were rated probably loose. PMID- 15118841 TI - Comparison of wear and osteolysis in hip replacement using two different coatings of the femoral stem. AB - We compared the clinical and radiographic results of two matched series of total hip arthroplasties, one with hydroxyapatite-coated femoral stems, the other with a similar but porous-coated femoral stem. The prevalence of radiographic osteolysis was 16% in hips with hydroxyapatite-coated stems and 43% in hips with porous-coated femoral stems. In hips with hydroxyapatite-coated stems, osteolysis was always limited to Gruen zones 1 and 7. In contrast, distal osteolysis was present around 26% of the porous-coated stems. At 7 years, the survival-free rate of distal osteolysis was 100% in hips with hydroxyapatite-coated stems but 90% in hips with porous-coated stems (p=0.04). Circumferential hydroxyapatite coating of the femoral component reduced the occurrence of osteolysis and eliminated distal osteolysis at 5-10 years of follow-up. In addition, hydroxyapatite coating did not alter the wear rate. PMID- 15118842 TI - Down-regulation of zeta-chain expression in T cells: a biomarker of prognosis in cancer? AB - The zeta chain is a 16-kDa molecule associated with the T-cell receptor (TCR)-CD3 complex in T lymphocytes and FcgammaRIII in CD3(-)CD56(+)CD16(+) natural killer (NK cells). The zeta chain functions as a transmembrane signaling molecule in lymphocytes. Expression of zeta was found to be decreased in CD4(+) and CD8(+) T lymphocytes isolated from the tumor site or from the peripheral circulation of patients with cancer. A quantitative flow cytometry-based assay for zeta-chain expression allows for reproducible serial evaluations of disease- or therapy associated changes in expression of this signaling molecule in phenotypically defined subsets of immune cells. Semiquantitative evaluation of zeta expression in paraffin-embedded tissue specimens can link it to the conventional markers of prognosis or survival. Several distinct mechanisms may be responsible for decreased/absent zeta in T cells of patients with cancer. Monitoring for zeta expression is useful for assessing immune competence in these patients and for following changes in immune competence during anticancer therapies. Correlations made between clinical findings, pathologic results, and zeta expression in immune cells suggest that low/absent zeta is predictive of poor prognosis and survival in patients with cancer. Thus, zeta is emerging as a clinically relevant signaling molecule, which also seems to predict a favorable response to biologic therapies and could be helpful in a selection of patients for immunotherapy trials. Validation studies have yet to be performed for this putative immunologic biomarker. Its consistent use for monitoring under standardized conditions of cancer patients treated with biotherapies may help in confirming a role for zeta as a correlate of prognosis or survival. PMID- 15118843 TI - 18FDG imaging of giant cell arteritis: usefulness of whole-body plus brain PET. PMID- 15118844 TI - MicroPET imaging of brain tumor angiogenesis with 18F-labeled PEGylated RGD peptide. AB - We have previously labeled cyclic RGD peptide c(RGDyK) with fluorine-18 through conjugation labeling via a prosthetic 4-[18F]fluorobenzoyl moiety and applied this [18F]FB-RGD radiotracer for alphav-integrin expression imaging in different preclinical tumor models with good tumor-to-background contrast. However, the unfavorable hepatobiliary excretion and rapid tumor washout rate of this tracer limit its potential clinical applications. The aims of this study were to modify the [18F]FB-RGD tracer by inserting a heterobifunctional poly(ethylene glycol) (PEG, M.W. =3,400) between the 18F radiolabel and the RGD moiety and to test this [18F]FB-PEG-RGD tracer for brain tumor targeting and in vivo kinetics. [18F]FB PEG-RGD was prepared by coupling the RGD-PEG conjugate with N-succinimidyl 4 [18F]fluorobenzoate ([18F]SFB) under slightly basic conditions (pH=8.5). The radiochemical yield was about 20-30% based on the active ester [18F]SFB, and specific activity was over 100 GBq/micromol. This tracer had fast blood clearance, rapid and high tumor uptake in the subcutaneous U87MG glioblastoma model (5.2+/-0.5%ID/g at 30 min p.i.). Moderately rapid tumor washout was observed, with the activity accumulation decreased to 2.2+/-0.4%ID/g at 4 h p.i. MicroPET and autoradiography imaging showed a very high tumor-to-background ratio and limited activity accumulation in the liver, kidneys and intestinal tracts. U87MG tumor implanted into the mouse forebrain was well visualized with [18F]FB PEG-RGD. Although uptake in the orthotopic tumor was significantly lower (P<0.01) than in the subcutaneous tumor, the maximum tumor-to-brain ratio still reached 5.0+/-0.6 due to low normal brain background. The results of H&E staining post mortem agreed with the anatomical information obtained from non-invasive microPET imaging. In conclusion, PEGylation suitably modifies the physiological behavior of the RGD peptide. [18F]FB-PEG-RGD gave improved tumor retention and in vivo kinetics compared with [18F]FB-RGD. PMID- 15118845 TI - Scatter and attenuation correction changes interpretation of gated myocardial perfusion imaging. AB - Attenuation correction may improve the diagnostic accuracy of myocardial perfusion imaging (MPI). However, few studies have dealt with the clinical consequences for reporting. We compared routine reports based on scatter corrected MPI (MPI-routine) with consensus readings of scatter-corrected (MPI scatter) and scatter plus attenuation-corrected studies (MPI-attenuation) to investigate the impact of attenuation correction on reporting. One hundred consecutive stable angina patients (including 55 men) were investigated in a 99mTc-sestamibi 2-day gated protocol with scatter and attenuation correction. With MPI-routine, 53 patients had normal perfusion and 47 abnormal perfusion, compared to 62 and 38 with MPI-attenuation, and 54 and 46, respectively, with MPI scatter. Agreement between MPI-routine and MPI-attenuation with respect to overall diagnosis (normal/abnormal perfusion) was 89% (kappa=0.78) compared to 95% (kappa=0.90) between MPI-routine and MPI-scatter. With MPI-attenuation, the overall routine diagnosis changed in 11 patients, of which ten cases were judged normal after scatter plus attenuation correction. The majority of the "normalised" studies were among patients with apparently single-vessel RCA disease as judged from MPI. Agreement rates with regard to normal, reversible or irreversible defects between MPI-attenuation and MPI-routine for the LAD, LCX and RCA territories were 88%, 97% and 85%, respectively, without significant sex differences. In conclusion, attenuation correction caused a change in diagnosis in approximately 10% of the patients, corresponding to one-fifth of the abnormal studies. In all but one case, the shift was from abnormal to normal, mostly because of a different interpretation in the RCA territory. PMID- 15118846 TI - 186Re-HEDP for metastatic bone pain in breast cancer patients. AB - Two-thirds of patients with metastatic cancer suffer from pain. Pain originating from skeletal metastases is the most common form of cancer-related pain. Bone pain, often exacerbated by pressure or movement, limits the patient's autonomy and social life. Pain palliation with bone-seeking radiopharmaceuticals has proven to be an effective treatment modality in patients with metastatic bone pain. These bone-seeking radiopharmaceuticals are extremely powerful in treating scattered painful bone metastases, for which external beam radiotherapy is impossible because of the large field of irradiation. (186)Re-hydroxyethylidene diphosphonate (HEDP) is a potentially useful radiopharmaceutical for this purpose, having numerous advantageous characteristics. Bone marrow toxicity is limited and reversible, which makes repetitive treatment safe. Studies have shown encouraging clinical results of palliative therapy using (186)Re-HEDP, with an overall response rate of ca. 70% in painful bone metastases. It is effective for fast palliation of painful bone metastases from various tumours and the effect tends to last longer if patients are treated early in the course of their disease. (186)Re-HEDP is at least as effective in breast cancer patients with painful bone metastases as in patients with metastatic prostate cancer. It is to be preferred to radiopharmaceuticals with a long physical half-life in this group of patients, who tend to have more extensive haematological toxicity since they have frequently been pretreated with bone marrow suppressive chemotherapy. This systemic form of radionuclide therapy is simple to administer and complements other treatment options. It has been associated with marked pain reduction, improved mobility in many patients, reduced dependence on analgesics, and improved performance status and quality of life. PMID- 15118847 TI - Degradation pathways of cyclic alkanes in Rhodococcus sp. NDKK48. AB - The degradation pathways for cyclic alkanes (c-alkanes) in Rhodococcus sp. NDKK48 were investigated. Strain NDKK48 used dodecylcyclohexane as a sole carbon and energy source, and five metabolites in the dodecylcyclohexane degradation pathway were detected by gas-chromatography/mass spectra. The metabolites were identified as cyclohexanecarboxylic acid, cyclohexylacetic acid, 1-cyclohexene-1-acetic acid, 4-dodecylcyclohexanol, and 4-dodecylcyclohexanone. The strain degrades dodecylcyclohexane via a ring oxidation pathway and an alkyl side chain oxidation pathway. Cyclohexanecarboxylic acid was further oxidized to muconic acid via 1 cyclohexene-1-carboxylic acid and benzoic acid, and the muconic acid was finally used by strain NDKK48 for growth. Methylcyclohexane and cyclohexane were co oxidized with hexadecane by strain NDKK48. Methylcyclohexane was degraded via a ring oxidation pathway, and the degradation pathway contained part of the Baeyer Villiger oxidation for ring cleavage. Cyclohexane was also degraded by the same pathway as methylcyclohexane. Thus, strain NDKK48 has two pathways for the complete degradation of c-alkanes. PMID- 15118848 TI - Genetic and environmental factors influencing the human factor H plasma levels. AB - Factor H is a plasma protein that plays a critical role in the regulation of complement activation in fluid phase and on cellular surfaces. Over the years numerous reports have illustrated the association of factor H deficiencies with chronic renal and infectious diseases. Plasma levels of factor H show a five-fold range of variation in humans (116-562 microg/ml), which may also be relevant to disease susceptibility. To quantify the effects of the genetic and environmental factors responsible for the variation in the factor H plasma levels, we have applied variance-component methods to a family-based sample. Factor H plasma levels show an age-dependent increase ( P<0.0001) and are decreased in smokers ( P<0.0001). Interestingly, the heritability of the factor H trait is very high ( h(2)=0.62+/-0.07; P<0.0001), indicating that 62% of the factor H phenotypic variance is due to the additive effects of genes. On this premise, we conducted a genome-wide linkage screen in order to identify genes regulating the factor H trait. Three genomic regions (1q32, 2p21-24 and 15q22-24) provided suggestive evidence of linkage (LOD scores 2.03, 2.15 and 2.00, respectively) with the plasma levels of factor H. PMID- 15118849 TI - Sequence and mRNA expression of nonclassical SLA class I genes SLA-7 and SLA-8. AB - Given the prominent position of pig endothelial cells in pig-to-human xenotransplantation and the role of classical and nonclassical MHC class I proteins in T and NK cell recognition, the expression of pig MHC (SLA) class I genes in a pig aortic endothelial cell line (AOC cells) was examined. Using a primer corresponding to a highly conserved region of exon 4, RT-PCR analysis of SLA class I expression in AOC cells revealed not only expression of the classical SLA class I ( SLA-1, -2, and -3) genes, but also SLA class I transcripts corresponding to SLA nonclassical class I (class Ib) genes SLA-6 and SLA-8. Further analysis of SLA class Ib expression in porcine aortic endothelial cells using SLA class I gene-specific primers confirmed SLA-6 and SLA-8 expression and also demonstrated expression of SLA-7. While SLA-6 has been relatively well characterized, no data regarding bona fide SLA-7 and SLA-8 transcripts have been reported. Therefore, cDNAs containing the complete open reading frames of SLA-6, 7, and -8 were obtained. Compared to an SLA-1 protein sequence, the predicted SLA 7 and -8 protein sequences exhibited most sequence divergence in alpha1, alpha2, and cytoplasmic domains. Expression of SLA-6, -7, and -8 was examined by RT-PCR using RNA prepared from a variety of tissues. SLA-6 transcripts were detected in every tissue examined. Except for brain, SLA-8 transcripts were similarly widespread. SLA-7 exhibited more limited tissue distribution. PMID- 15118850 TI - A single amino-acid polymorphism in pocket A of HLA-A*6602 alters the auxiliary anchors compared with HLA-A*6601 ligands. AB - In this study we have sequenced peptides eluted from a truncated recombinant HLA A*6602 molecule, and compared their features with data reported for peptides presented in the A*6601 molecule. A striking change in the amino-acid binding preferences was observed at peptide position P1, which interacts with pocket A of the HLA peptide-binding region. For A*6601, aspartic acid and glutamic acid, both of which possess polar acidic side-chains, have been described as auxiliary anchors. This is in marked contrast to A*6602, where we observed serine, which has a neutral polar side-chain, as auxiliary anchor at P1. Accordingly, this shift in the physico-chemical properties of the auxiliary anchor may be best explained by the HLA amino-acid polymorphism at position 163, where arginine (hydrophilic, alkaline) in A*6601 has been replaced by glutamic acid in A*6602. This amino-acid exchange results in a shift towards higher acidity in pocket A, apparently resulting in the loss of preference for acidic auxiliary anchors, and leading to the preference for the neutral amino acid serine. The change of the auxiliary anchor residue at P1 is likely to alter the spectrum of peptides presented by A*6602 compared with A*6601, which may result in allogenicity in the case of a mismatch in allogeneic stem cell transplantation. PMID- 15118851 TI - Mapping of a new quantitative trait locus for resistance to malaria in mice by a comparative mapping approach with human Chromosome 5q31-q33. AB - A number of linkage studies in human populations have identified a locus ( pfbi) on Chromosome 5q31-q33 controlling Plasmodiun falciparum blood infection levels. This region contains numerous candidate genes encoding immunological molecules such as cytokines, growth factors and growth-factor receptors. We have used an F(11) advance intercross line (AIL) population of mice infected with Plasmodium chabaudi to identify additional mouse quantitative trait loci (QTL) for control of parasitaemia on Chrs 11 and 18, which carry regions homologous to human Chr 5q31-q33. Herein, we report a novel QTL for parasitaemia control ( char8) on the mouse Chr 11, linked to marker D11Mit242, and involved in the clearance stages of the parasites from the bloodstream. Strikingly, several Th2 cytokines that are located within char8 have been identified to play a predominant role in the late stages of the infection. PMID- 15118852 TI - Comparative analysis of different apoptosis detection methods in human testicular cancer. AB - In situ end-labeling (ISEL) of internucleosomal 3' DNA strand breaks and the morphological proof of nuclear chromatin condensation are two widely used methods to investigate and quantify apoptosis. However, it is still unclear whether both processes are linked with each other and if quantifying apoptosis by both methods leads to comparable results. Therefore, internucleosomal DNA fragmentation and chromatin condensation were measured simultaneously on double-fluorescence labeled sections of 62 testicular tumors (47 nonseminomatous tumors and 15 seminomas) using immunofluorescence microscopy. Different apoptotic indices (AI), based on DNA fragmentation and/or morphological criteria were determined. The AI were quantified. Morphologically obtained AI ranged between 1.99% for non seminomatous tumors and 0.88% for seminomas. The detection of DNA fragmentation values ranged between 8.15% for non-seminomatous tumors and 2.70% for seminomas. Only about 30% of all apoptotic cells could be detected with the morphological method compared to 80% using ISEL in both tumor entities. Therefore, the equivalence of investigations using different apoptosis detection methods in human testicular cancer seems questionable. PMID- 15118853 TI - Gastrointestinal stromal tumor: report of two unusual cases. PMID- 15118854 TI - Reduction of PTEN and p27kip1 expression correlates with tumor grade in prostate cancer. Analysis in radical prostatectomy specimens and needle biopsies. AB - The extreme variability of prostate cancer implies latent disease with missing clinical symptoms in some cases. Tumor suppressors PTEN (phosphatase and tensin homolog deleted on chromosome ten) and p27kip1 are frequently mutated in various human cancers. PTEN negatively influences cell growth and induces apoptosis, while p27kip1 binds to cyclin-E-Cdk2 and counteracts mitosis. This study investigated the expression of PTEN and p27kip1 in prostatectomies and needle biopsies in order to determine whether protein localization or expression levels are correlated with tumor grade and whether PTEN and p27kip1 expression in biopsies are valuable predictive tumor markers. Analysis of PTEN demonstrated that weak expression levels were significantly more prevalent in high-grade tumors. Analysis of p27kip1 revealed that high-grade tumors had a higher percentage of cytoplasmic localization of the protein than low-grade tumors, where nuclear localization was more frequent. Furthermore, this study indicated a positive association between PTEN and p27kip1 levels. An increase of high-grade tumors corresponded to a progressive loss of both tumor suppressors in needle biopsies and prostatectomies. p27kip1 and PTEN did not show a higher predictive accuracy of the tumor grade in the surgical specimen than the Gleason score. However, p27kip1 had the same predictive value as the Gleason score in needle biopsies. PMID- 15118855 TI - New cell lines with chondrocytic phenotypes from human chondrosarcoma. AB - In the present study, we investigated chondrocytic characterization for newly established human chondrosarcoma cell lines. A chondrosarcoma cell line, HCS-TG, was established by the implantation of grade-2 human chondrosarcoma into athymic mice. Cloning of HCS-TG cells from passage 17 was performed. After cell cloning, two clonal-cell lines (HCS-TG C3 and E2) with good proliferative activities were obtained. These cell lines in monolayer culture retained a polygonal morphology. Their doubling times were 68 h and 45 h, respectively. mRNA expressions of type I, -II, -X, and -XI collagens and aggrecan core protein were detected on reverse transcription polymerase chain reaction. Protein expression of type-II collagen was confirmed in each cell line using Western blotting. However, there was no expression of type-I collagen. Moreover, gelatin zymography revealed that both cell lines produced extracellular matrices with matrix metalloproteinases 2 and 9. The parental HCS-TG cells had tumorigenicity in athymic mice; however, C3 and E2 were not tumorigenic. New clonal-cell lines HCS-TG C3 and E2 derived from human chondrosarcoma are morphologically chondrocytic in serial monolayer cultures and express chondrocytic phenotypes. PMID- 15118857 TI - Mannan transglycosylase: a novel enzyme activity in cell walls of higher plants. AB - Mannan transglycosylase is a novel cell wall enzyme activity acting on mannan based plant polysaccharides in primary cell walls of monocotyledons and dicotyledons. The enzyme activity was detected by its ability to transfer galactoglucomannan (GGM) polysaccharides to tritium-labelled GGM-derived oligosaccharides generating tritium-labelled GGM polysaccharides. Mannan transglycosylase was found in a range of plant species and tissues. High levels of the enzyme activity were present in flowers of some kiwifruit (Actinidia) species and in ripe tomato (Solanum lycopersicum L.) fruit. Low levels were detected in mature green tomato fruit and activity increased during tomato fruit ripening up to the red ripe stage. Essentially all activity was found in the tomato skin and outermost 2 mm of tissue. Mannan transglycosylase activity in tomato skin and outer pericarp is specific for mannan-based plant polysaccharides, including GGM, galactomannan, glucomannan and mannan. The exact structural requirements for valid acceptors remain to be defined. Nevertheless, a mannose residue at the second position of the sugar chain and the absence of a galactose substituent on the fourth residue (counting from the non-reducing end) appear to be minimal requirements. Mannan-based polysaccharides in the plant cell wall may have a role analogous to that of xyloglucans, introducing flexibility and forming growth-restraining networks with cellulose. Thus mannan transglycosylase and xyloglucan endotransglycosylase, the only other known transglycosylase activity in plant cell walls, may both be involved in remodelling and refining the cellulose framework in developmental processes throughout the life of a plant. PMID- 15118858 TI - Expression of the patatin-related phospholipase A gene AtPLA IIA in Arabidopsis thaliana is up-regulated by salicylic acid, wounding, ethylene, and iron and phosphate deficiency. AB - In Arabidopsis thaliana (L.) Heynh., the cytosolic, patatin-related phospholipase A enzymes comprise a family of ten genes designated AtPLAs thought to be involved in auxin and pathogen signalling [A. Holk et al. (2002) Plant Physiol 130:90 101]. One of these, AtPLA IIA, is investigated here by studying its transcriptional regulation through transgenic Arabidopsis plants containing the AtPLA IIA promoter (PIIA) fused to the beta-glucuronidase (GUS) gene. GUS activity appeared in leaves at 10-12 days and became increasingly stronger with age in all leaves. From the same age on, strong GUS activity was visible in the basal stipules of the rosette leaves. PIIA-dependent GUS activity was found in the older parts of the primary root (from 10 days on) and, later in development, in older parts of side roots, and the root cap. No GUS activity was detected in flower organs. PIIA-dependent GUS expression in 12-day-old plants was up regulated after treatment by salicylic acid, Bion, wounding, 1-aminocyclopropane 1-carboxylic acid (ACC) and jasmonic acid. When transgenic PIIA:: uidA plants were grown devoid of iron, 9-day-old plants exhibited increased GUS activity in the leaves and, when devoid of phosphate, 11-day-old plants had increased GUS activity in the roots. In conclusion, this member of the patatin-related phospholipase A gene family showed properties of a defence and iron-stress and phosphate-stress gene, being transcriptionally up-regulated within hours or days. PMID- 15118859 TI - Interactions of abscisic acid and sugar signalling in the regulation of leaf senescence. AB - Leaf senescence can be triggered by a high availability of carbon relative to nitrogen or by external application of abscisic acid (ABA). Most Arabidopsis mutants with decreased sugar sensitivity during early plant development are either ABA insensitive (abi mutants) or ABA deficient (aba mutants). To analyse the interactions of carbon, nitrogen and ABA in the regulation of senescence, wild-type Arabidopsis thaliana (L.) Heynh. and aba and abi mutants were grown on medium with varied glucose and nitrogen supply. On medium containing glucose in combination with low, but not in combination with high nitrogen supply, senescence was accelerated and sucrose, glucose and fructose accumulated strongly. In abi mutants that are not affected in sugar responses during early development (abi1-1 and abi2-1), we observed no difference in the sugar-dependent regulation of senescence compared to wild-type plants. Similarly, senescence was not affected in the sugar-insensitive abi4-1 mutant. In contrast, the abi5-1 mutant did exhibit a delay in senescence compared to its wild type. As ABA has been reported to induce senescence and ABA deficiency results in sugar insensitivity during early development, we expected senescence to be delayed in aba mutants. However, the aba1-1 and aba2-1 mutants showed accelerated senescence compared to their wild types on glucose-containing medium. Our results show that, in contrast to sugar signalling in seedlings, ABA is not required for the sugar dependent induction of leaf senescence. Instead, increased sensitivity to osmotic stress could have triggered early senescence in the aba mutants. PMID- 15118860 TI - mRNA expression of fibroblast growth factors and hepatocyte growth factor in rat plantaris muscle following denervation and compensatory overload. AB - We addressed the question of whether hypertrophy induced by compensatory overload differs according to innervation status, and how fibroblast growth factors (FGF) and hepatocyte growth factor (HGF) mRNAs are expressed in the rat plantaris muscle during overload (OL) and/or denervation. Male Wistar rats were divided into four groups (Normal-Cont, Normal-OL, Denervated-Cont, and Denervated-OL). according to the plantaris denervation and/or overload. Three weeks later, plantaris weight in Denervated-Cont and Denervated-OL was significantly lower than in the Normal-Cont. The muscle weights in the Normal-OL were higher than in the Normal-Cont. The muscle weights in the Denervated-OL were higher than in the Denervated-Cont. Three days after the treatment, FGF-2, FGF-6, FGF-7 and HGF mRNAs in the Normal-OL were significantly higher than those in the Normal-Cont. FGF-2, FGF-6, FGF-7 and HGF mRNAs in the Denervated-OL were also significantly higher after 3 days than those in the Denervated-Cont. After 7 days, FGF-2, FGF 5, FGF-6, FGF-7 and HGF mRNAs were significantly higher in the Normal-OL than those in the Normal-Cont. At 21 days, FGF-1, FGF-6 and HGF mRNA levels were significantly increased. In the Denervated-OL, FGF-2, FGF-7 and HGF mRNAs at 7 days, and FGF-2 mRNA at 21 days were significantly higher than those in the Denervated-Cont. FGF-2 and FGF-6 mRNA levels decreased significantly following denervation; however, FGF-1, FGF-5, FGF-7 and HGF mRNA levels increased and maintained this increase for the 21-days treatment period. Muscle hypertrophy was thus induced by compensatory overload irrespective of innervation status, possibly in association with certain FGFs and HGF. The differential mRNA expression patterns of FGFs and HGF observed following compensatory overload and/or denervation suggest distinct roles for individual FGFs and HGF in muscle hypertrophy and/or atrophy. PMID- 15118862 TI - Expert assessment of physical ergonomics at video-display unit workstations: repeatability, validity and responsiveness to changes. AB - AIMS: Inter-observer repeatability, validity and responsiveness to change were determined for an expert assessment method for video-display unit (VDU) workstation ergonomics. The aim was to determine to what extent the expert assessment of ergonomics is related to the technical measurements, tidiness and space, work chair ergonomics and responds to changes in these characteristics. METHODS: Technical measurements and video-recordings before and 2 months after an ergonomic intervention were made for 109 VDU office workstations. Two experts in ergonomics analysed and rated the ergonomics of the workstations. A researcher analysed tidiness and available space. A physiotherapist classified the work chairs used according to their ergonomic properties. RESULTS: The intra-class correlation coefficient between the workstation ergonomic ratings of the two experts was 0.74 at the baseline and 0.81 at the follow-up. Workstation tidiness and space, and work chair ergonomics, had a strong effect on the assessments of both experts. For both experts a change in the locations of the mouse, the screen and the keyboard and values of tidiness and space and work chair ergonomics during the intervention showed a significant association with the ratings. CONCLUSION: The assessment method studied can be utilized by an expert in a repeatable manner both in cross-sectional and in longitudinal settings. PMID- 15118861 TI - Volume replacement and microhemodynamic changes in polytrauma. AB - Though fluid administration is one of the most basic concepts in resuscitation, there is ongoing controversy and continuing research on the definition of the ideal fluid for resuscitation of trauma and hemorrhage and for intraoperative volume support. In general, crystalloids and colloids, as well as blood, blood substitutes and oxygen therapeutics, are available. This report briefly revisits the physiological mechanisms underlying resuscitation with crystalloids and colloids, emphasizing colloid-supplemented resuscitation with hypertonic saline. Finally, potential applications of oxygen therapeutics are briefly considered. PMID- 15118864 TI - Dosimetric response of tooth enamel to 14 Mev neutrons. AB - The electron paramagnetic resonance (EPR) response of tooth enamel in a monenergetic neutron beam of 14 MeV was studied, with the aim to evaluate the relative neutron to (60)Co sensitivity. Three samples of tooth enamel powder were irradiated in air. A whole tooth and a powdered sample were irradiated in a geometrical PMMA phantom, in order to simulate the real exposure of a tooth inside a human head. The measured dose in enamel was compared to the dose calculated by Monte Carlo simulation. The relative neutron to (60)Co sensitivity using different reference materials (air, water and enamel) was evaluated as well. Large differences in sensitivity values were found depending on the reference material: the obtained relative neutron to (60)Co sensitivity was 0.47+/-0.09 for enamel and 0.15+/-0.03 for water. A comparison with results in fast neutron fields is reported. PMID- 15118874 TI - Population-based study on incidence, survival rates, and genetic alterations of low-grade diffuse astrocytomas and oligodendrogliomas. AB - We carried out a population-based study on low-grade diffuse gliomas in the Canton of Zurich, Switzerland (population 1.16 million). From 1980 to 1994, 987 astrocytic and oligodendroglial tumors were diagnosed, of which 122 (12.4%) were low-grade (WHO grade II). The incidence rates adjusted to the World Standard Population, per million population per year, were 2.28 for low-grade diffuse astrocytomas, 0.89 for oligoastrocytomas, and 2.45 for oligodendrogliomas. The survival rate (mean follow-up 7.5+/-4.8 years) was highest for patients with oligodendroglioma (78% at 5 years, 51% at 10 years), followed by those with oligoastrocytoma (70% at 5 years, 49% at 10 years) and fibrillary astrocytoma (65% at 5 years, 31% at 10 years). Survival of patients with gemistocytic astrocytoma was poor, with survival rates of 16% at 5 years and 0% at 10 years. Younger patients (<50 years) survived significantly longer than older patients (>50 years; P=0.013). DNA sequencing, performed in 84% of cases, revealed that TP53 mutations were most frequent in gemistocytic astrocytomas (88%), followed by fibrillary astrocytomas (53%) and oligoastrocytomas (44%), but were infrequent (13%) in oligodendrogliomas. The presence of TP53 mutations was associated with shorter survival of patients with low-grade diffuse gliomas (log-rank test; P=0.047), but when each histological type was analyzed separately, an association was observed only for oligoastrocytoma ( P=0.05). Loss on 1p and 19q were assessed by quantitative microsatellite analysis in 67% of cases. These alterations were frequent in oligodendrogliomas (1p, 57%; 19q, 69%), less common in oligoastrocytomas (1p, 27%; 19q, 45%), rare in fibrillary astrocytomas (1p, 7%; 19q, 7%), and absent in gemistocytic astrocytomas. None of these alterations were predictive of survival. These results establish the frequency of key genetic alterations in low-grade diffuse gliomas at a population-based level. Multivariate Cox's regression analysis indicates that only age and histological type, but not genetic alterations, are significant predictive factors. PMID- 15118875 TI - Lung contusion-lacerations after blunt thoracic trauma in children. AB - Chest trauma in children is an indicator of injury severity and is associated with a high mortality rate. The aim of this study was to investigate the impact of pulmonary contusion-laceration on short and long-term outcome of pediatric patients after blunt thoracic trauma. A retrospective analysis of records of 41 children aged 10 months to 17 years who were treated for pulmonary and associated injuries between 1986 and 2000 was done concerning mode of injury, types of injuries, management and outcome. In addition, a follow-up investigation was performed 4.5+/-1 years after injury. Of the patients 27 were involved in motor vehicle accidents (MVA group) and 14 patients suffered other types of accidents (others group). The mean injury severity score (ISS) was 30+/-2 (range 9-75) with no significant difference between the groups. Patients from the MVA group suffered more frequently bilateral pulmonary lesions and needed more often chest tube placement ( p<0.05), 5 patients died (12%) all from the MVA group. The follow-up investigation of 34 patients showed unremarkable chest x-rays and normal lung function in all but 1 patient with bronchial asthma. In conclusion, children who recover after a pulmonary contusion-laceration trauma do not suffer from significant late respiratory problems. PMID- 15118876 TI - Hairy polyp of the tongue: a case report. AB - Hairy polyps or dermoids of the oro- and nasopharynx are benign lesions containing elements of both ectodermal and mesodermal origin. Because of its rarity, we report a case of hairy polyp arising from the tongue in a 40-day-old infant. The lesion was covered by squamous epithelium and a central core of fibroadipose tissue, minor salivary glands, and cartilage. We discuss the clinicopathological features, terminology, etiology, and differential diagnosis of this condition. PMID- 15118877 TI - Molecular aspects of the interaction between amphotericin B and a phospholipid bilayer: molecular dynamics studies. AB - Amphotericin B (AmB) is a polyene macrolide antibiotic used to treat systemic fungal infections. The molecular mechanism of AmB action is still only partly characterized. AmB interacts with cell-membrane components and forms membrane channels that eventually lead to cell death. The interaction between AmB and the membrane surface can be regarded as the first (presumably crucial) step on the way to channel formation. In this study molecular dynamics simulations were performed for an AmB-lipid bilayer model in order to characterize the molecular aspects of AmB-membrane interactions. The system studied contained a box of 200 dimyristoylphosphatidylcholine (DMPC) molecules, a single AmB molecule placed on the surface of the lipid bilayer and 8,065 water molecules. Two molecular dynamics simulations (NVT ensemble), each lasting 1 ns, were performed for the model studied. Two different programs, CHARMM and NAMD2, were used in order to test simulation conditions. The analysis of MD trajectories brought interesting information concerning interactions between polar groups of AmB and both DMPC and water molecules. Our studies show that AmB preferentially took a vertical position, perpendicular to the membrane surface, with no propensity to enter the membrane. Our finding may suggest that a single AmB molecule entering the membrane is very unlikely. PMID- 15118878 TI - Recruitment of divalent metal ions by incorporation of 4-thio-2'-deoxythymidine or 4-thio-2'-deoxyuridine into DNA. AB - The modified nucleosides 4-thio-2'-deoxyuridine (s4dU) and 4-thio-2' deoxythymidine (s4dT) are incorporated into dinucleosides, and s4dT is incorporated into a DNA hairpin loop to provide divalent metal ion binding sites. Binding of two different metal ions to these sites is studied, including Cd(II) as an NMR spectroscopy probe and Cu(II) as a reactive metal ion for DNA cleavage. Binding of Cd(II) to 4-thiouridine (s4U) and s4dT nucleosides, s4dU- and s4dT containing dinucleosides, and a hairpin loop oligonucleotide containing s4dT is monitored by following the change in UV-vis absorbance of the thionucleosides at 340 nm and 21 degrees C in solutions containing 20.0-40 mM buffer, 1.00 M NaCl, and 15.0 mM BaCl2. Cd(II) binds to the N3 deprotonated form of s4dT with a binding constant (K = 1.1 x 10(4) M(-1)) that is similar to that for Cd(II) binding to d(Tps4T) (K = 9.2 x 10(3) M(-1)). Apparent binding constants (Kapp) at pH 7.7 of Cd(II) to dinucleosides d(Gps4T), d(s4TpG), and d(Gps4U) are similar to those of their respective nucleosides s4U and s4dT, suggesting that neither the phosphate diester nor the second nucleoside has a major effect on Cd(II) binding. Binding of Cd(II) to s4U and d(Gps4U) is studied by use of 113Cd NMR and 1H NMR spectroscopy, respectively. Binding strength and stoichiometry of the Cd(II) complex with d(Gps4U) as studied by 1H NMR spectroscopy are similar to that obtained by UV-vis spectroscopy. Cd(II) binds strongly to s4dT in the loop portion of a DNA hairpin loop (Kapp = 2.7 x 10(3) M(-1) at pH 7.7). However, the hairpin loop is moderately destabilized by Cd(II) binding, with a decrease in T(m) of 14 degrees C in the presence of 10.0 mM Cd(II) as determined by optical melting experiments. Cu(II) oxidizes s4dT to form the disulfide of s4dT, limiting the usefulness of further studies with Cu(II). PMID- 15118879 TI - The impact of tumour volume and surgery on the outcome of adults with supratentorial WHO grade II astrocytomas and oligoastrocytomas. AB - BACKGROUND: The study was performed to elucidate the impact of tumour volume and surgical resection on the long-term outcome of patients with supratentorial, diffuse, World Health Organization (WHO) grade II astrocytomas and oligo astrocytomas. METHOD: After analysing 79 adult patients consecutively diagnosed between 1991 and 2000, we selected a group of 42 patients treated by surgery without adjuvant therapy. The tumour volume was defined as the whole region of T2 hyperintensity and measured interactively on pre- and postoperative and follow-up Magnetic Resonance Imaging (MRI) using a dedicated imaging software. Volumetric, clinical, and histological data were analysed for correlation with tumour progression (TP), malignant transformation (MT), drop in functional status (DKPS) and overall survival (OS). FINDINGS: Pre- and postoperative tumour volumes, and the involvement of more than one lobe were strongly associated with worse outcome. Preoperative tumour volume was the strongest predictor of OS (p<0.01) and the only predictor of MT (p<0.05). The absolute and relative volumes of tumour removed by surgery were not significantly associated with outcome. CONCLUSIONS. Initial tumour volume, measured as the volume of T2-hyperintensity on MRI, and tumour extension are the strongest predictors of outcome in patients with supratentorial diffuse astrocytic WHO Grade II tumours. The potential benefit of aggressive tumour resection needs to be investigated in a prospective controlled trial. PMID- 15118880 TI - Protein S-100b as serum marker for prediction of functional outcome in metastatic spinal cord compression. AB - BACKGROUND: To evaluate the significance of protein S-100b as a serum marker for the prediction of functional outcome in the event of symptomatic spinal cord compression due to epidural metastases. METHOD: 34 patients with paresis due to metastatic spinal cord compression were included in this prospective study. Venous blood samples for protein S-100b were taken after admission and regularly after operative decompression. The individual time course of protein S-100b levels was correlated with the clinical outcome by means of motor function. Outcome was considered to be favourable in case of neurological improvement and preservation or retrieval of walking ability whereas non-improvement or further neurological deterioration without restoration of function of ambulation was regarded to be unfavourable. FINDINGS: Patients with favourable outcome had serum levels of S-100b which were either normal all the time or which were initially increased but normalised within 2 to 3 days. Patients with unfavourable outcome, however, had increased levels throughout which showed either a further increase or only a slow decrease within approximately two weeks (p=0.0001). INTERPRETATION: These preliminary results suggest that, analogous to cerebral disorders, protein S-100b might be a promising serum marker to predict functional outcome in symptomatic spinal cord compression. PMID- 15118881 TI - Discharge criteria from intensive care unit in brain injured patients. AB - OBJECTIVE: We investigated the value of information on clinical features and intensity of treatment activity in the Intensive Care Unit (ICU) in predicting the need for further interventions after a patient is discharged from the Intensive Care Unit. Our aim was to assess if this could aid in making decisions about transfer to an Immediate Care Unit (sub-ICU). METHODS: We studied 39 patients with acute brain damage (traumatic or vascular causes). They ranged in age from 15 to 75 years and none had an associated spinal cord injury. The SAPS II, Glasgow Coma Scale (GCS), length of stay in ICU, duration of hospital stay and daily NEMS scores were recorded for each patient. We recorded the activities performed after transfer to an sICU, including complications that required active "life-saving" treatment. The role of each factor was assessed by using the odds ratio (OR), and with linear logistic regression. FINDINGS: 8 of the 39 patients developed a complication in the Sub-ICU. A linear logistic regression analysis demonstrated that the principal features having significant predictive value were:a) age, with an increase in risk of over 10 times for patients that were older than 50 (p=0.011);b) SAPS II scores > or = 50 points, with 24 times an increase in risk (p=0.002); and c) a GCS score < or =5 points, with an increase in risk of almost 7 times (p=0.024). INTERPRETATION: Complications in Sub-ICU are less likely in patients younger than 50 and who have SAPS II and GCS scores within predetermined limits. These indices can help in making decisions about discharge of a patient from ICU to Sub-ICU. PMID- 15118882 TI - Clinical predictors and neuropsychological outcome in severe traumatic brain injury patients. AB - BACKGROUND: The aim of the study was to evaluate the possible significant role of some clinical factors in predicting cognitive outcome in a group of severe traumatic brain injury (TBI) patients, with Glasgow Coma Scale (GCS) lower than 8 and duration of unconsciousness for at least 15 days (prolonged coma). METHOD: A consecutive sample of 25 survivors of severe TBI attending the Physical and Cognitive Rehabilitation program participated in this study. The neuropsychological test battery included: Word-list Learning, Prose recall, Rey Figure Delayed recall, Word fluency, Raven's Progressive Matrices' 47. The clinical variables evaluated in correlation with the neuropsychological outcome were the following: age, duration of unconsciousness, duration of post-traumatic amnesia, interval from head trauma to neuropsychological evaluation, interval from head trauma to recovery of oral feeding, and finally interval from head trauma to first verbal communication. FINDINGS: The clinical variable with a significant predictive value on most neuropsychological scores was the interval from head trauma to the recovery of oral feeding. CONCLUSIONS: If this result is confirmed in larger samples, time interval of oral feeding recovery from head trauma should be considered as a possible predictor of neuropsychological outcome in TBI patients with prolonged coma. PMID- 15118883 TI - Infected CNS infusion pumps. Is there a chance for treatment without removal? AB - BACKGROUND: Intrathecal baclofen administration by means of an implantable pump is nowadays a safe and effective method in the treatment of spasticity. One of the rare but devastating complications of this technique is pump infection, with a variety of Gram (-) and Gram (+) organisms being involved. Treatment of these infections, according to international literature, requires removal of the device and appropriate antibiotic therapy. METHOD: This article reports the authors experience in treating 3 patients with severe, medically intractable spasticity, suffering from infection of the intrathecally-delivering pump. A decision was made not to replace the device, but to treat this complication with pump disinfection and with a new treatment modality that has never been used before, the intra-pocket administration of antibiotics. FINDINGS: In all cases the infection was eradicated and the integrity of the pump maintained. None of the patients required a procedure under general anesthesia. CONCLUSIONS: Removal should no longer be considered the first treatment option in infections of intrathecally delivering pumps, especially those due to non-adherent bacteria, with mild clinical symptomatology. An initial attempt should always be made for conservative treatment. Intra-pocket administration of antibiotics helps in achieving high drugs levels locally, and may prove an important element in our armamentarium against such infections. PMID- 15118884 TI - Treatment of pain from osteoporotic vertebral collapse by percutaneous PMMA vertebroplasty. AB - BACKGROUND: Vertebral compression fractures are common complications in advanced osteoporosis. In general, this disease of the elderly patient is characterized by severe local back pain. Pathophysiologically, bony instability triggers local pain during body movement. Serious pain immobilizes the patients and forces them to bed rest. As a result, complications like thrombosis or pneumonia occur. Invasive treatment with surgical instrumentation for vertebral stabilization is not indicated in elderly patients especially with additional diseases. The purpose of this study was to test the hypothesis that percutaneous polymethylmethacrylate (PMMA) vertebroplasty significantly reduces pain due to vertebral collapse in osteoporotic patients and improves quality of life. METHODS: A total of 38 patients with osteoporotic vertebral compression fractures of the thoracic and lumbar spine were treated by PMMA vertebroplasty. After admission, before discharge from the hospital, six weeks, half a year and one year later patients answered the Oswestry Low Back Pain Disability (OLBPD) Questionnaire for assessment of treatment related change in disability. In all patients percutaneous vertebroplasty was performed under local anesthesia. FINDINGS: A total of 92% of patients reported a significant pain reduction immediately after treatment. Also one year after vertebroplasty pain remained significantly reduced. Vertebroplasty was highly beneficial for patients with pain related to local instability of the spine. Extravasation of PMMA beyond the vertebral margins was observed in 26% of the cases. No treatment related clinical or neurological complications were noticed. INTERPRETATION: PMMA vertebroplasty is a useful and safe method of pain relief which rapidly regains quality of life for patients with osteoporotic vertebral compression. PMID- 15118885 TI - Cell index--a new parameter for the early diagnosis of ventriculostomy (external ventricular drainage)-related ventriculitis in patients with intraventricular hemorrhage? AB - Temporary intraventricular catheters for managing acute obstructive hydrocephalus caused by intraventricular haemorrhage carry a high risk of developing ventriculostomy-related ventriculitis (VRV). The aim of this prospective study was to validate a new parameter for the early detection of an intraventricular infection. METHODS: Patients with external ventricular drainage due to intraventricular haemorrhage were enrolled in this prospective study. Leucocytes and erythrocytes in cerebrospinal fluid (CSF) and peripheral blood as well as bacteriological and chemical analysis of both were examined daily. The ratio of leucocytes to erythrocytes in CSF and leucocytes to erythrocytes in peripheral blood was calculated (so called cell index (CI)) and these values were compared with the "conventionally diagnosed" drain-associated ventriculitis. Furthermore, the CI values of the non-ventriculitis and ventriculitis group were compared using the t-test with adjustment for unequal variances (Welch test). RESULTS: Thirteen patients with an external ventricular drainage (EVD) expected to be in place for more than seven days were enrolled. Seven patients developed a bacteriologically proven VRV (time 0) within 12 days (mean 8.57). Diagnosis of VRV by CI was possible up to 3 days (mean 2.28) prior to conventional diagnosis. P values (Welch test) showed a significant difference on days -3 (P = 0.03), -2 (P = 0.03) and -1 (P = 0.012) - i.e. 3, 2 or 1 day, respectively, prior to the time point when the CSF culture grew staphylococci -, when compared with the mean cell indices of the controls, and a highly significant difference on time 0 (P < 0.001). CONCLUSION: The calculated CI allows the diagnosis of nosocomial VRV in patients with intraventricular haemorrhage at a very early point of time. PMID- 15118886 TI - Inhibition of the arachidonic acid metabolism blocks endothelial cell migration and induces apoptosis. AB - Previous studies have demonstrated that inhibitors of the arachidonic acid metabolism block migration and sensitise human glioma cells to treatment inducing apoptosis. This paradigm may provide a new concept for anti-invasive treatment strategies targeting invasive glioma cells. However, the effect of such treatment on other cellular elements in glial tumours such as endothelial cells is unknown. In this study we have analysed the expression of metabolites of the arachidonic acid pathway in endothelial cells in vitro and in vivo and we have assessed the influence of inhibitors of this pathway on motility, capillary like tube formation, and apoptosis in human endothelial cells. Human endothelial cells (HUVEC) in culture showed expression for thromboxane synthase and both isoforms of cyclo-oxygenase, COX-1 and COX-2. Immunostaining demonstrated low levels of COX-1 expression in capillaries and larger vessels of normal brain and moderately elevated levels of this enzyme in small vessels of brain tumours of various grades. Both thromboxane synthase and COX-2 expression was limited to endothelial cells found in anaplastic gliomas and glioblastomas. Thromboxane synthase inhibitors strongly decreased endothelial cell migration in HUVEC in vitro and capillary like tube formation was strongly inhibited by the compound at a similar dose range. The non-selective cyclo-oxygenase inhibitor ASA and the selective COX 2 inhibitor sulindac only had a minor effect on endothelial cell migration, however, the COX-2 inhibitor sulindac showed a synergistic effect with the thromboxane synthase inhibitor. Thromboxane synthase inhibitors induced apoptosis in endothelial cells as demonstrated by intracellular histone-complexed DNA fragmentation. These data suggest that inhibitors of thromboxane synthase influence migration and apoptosis in both human glioma cells and human endothelial cells. An anti-invasive treatment strategy using this class of compounds may therefore not only sensitise glioma cells to conventional treatments inducing apoptosis but may also be supported by an anti-angiogenic effect. PMID- 15118887 TI - Angiographic, hemodynamic and histological characterization of an arteriovenous fistula in rats. AB - BACKGROUND: Our understanding of the pathogenesis of arteriovenous malformations (AVMs) and arteriovenous fistulas (AVFs) has been limited by the lack of adequate animal models. In this study we evaluate the time course of angiographic, hemodynamic and histopathological changes in an arteriovenous fistula in rats as a potential model. METHODS: An arteriovenous fistula was created by a side-to-end anastomosis of the common carotid artery (CCA) to the external jugular vein (EJV). The animals underwent angiography of the fistula and were sacrificed 1, 7, 21, 42 or 90 days later. Flow and pressure measurements were performed in the CCA and ipsi- and contralateral EJV and detailed histological examination of whole mount sections of the fistula and cranium were done on fixed sections. Immunohistochemistry for CD31, smooth muscle alpha-actin and Ki-67 were performed. FINDINGS: Hemodynamic changes occur immediately after fistula formation creating a stable high flow, low resistant state. This induces a gradual increase in the inner diameter of the EJV and transverse sinus followed by a decrease in size of the transverse sinus. This decrease is associated with increased expression of alpha-actin in the wall of the sinus. The fistula becomes angiographically and histologically stable after 21 days. CONCLUSION: This model describes the time course of hemodynamic and histopathological changes after occur after AVF formation. Stabilization after 21 days makes it an attractive model for mechanistic and therapeutic studies of AVFs. PMID- 15118888 TI - Intradural en-bloc removal of the anterior clinoid process. AB - BACKGROUND: Anterior clinoidectomy is useful in the surgical treatment of paraclinoid and parasellar lesions. Previously reported procedures require expertise in drilling, the alternative method reported here reduces the drilling procedure. METHODS AND RESULTS: En-bloc clinoidectomy is performed intradurally via the standard pterional approach. A 1 to 2 mm-wide narrow drill line is placed with a 1 mm-wide diamond burr through the lesser sphenoid wing. It encircles the medial border of the optic canal and the lateral border of the lesser sphenoid wing over the superior orbital fissure, and is located about 1 cm anterior to the posterior margin of the optic canal. After drilling, one bony piece that includes the anterior clinoid process (ACP) and the optic canal roof remains connected to the basisphenoid bone by the optic strut alone. The optic strut is then fractured easily by applying leverage near its junction with the basisphenoid bone and the piece is removed en bloc with the major part of the optic strut, requiring little or no additional drilling of the residual bony fragments. Of 37 patients who underwent our en-bloc clinoidectomy, only one suffered complications consisting of cerebrospinal fluid (CSF) leakage through the sphenoid sinus. Our procedure requires an average of 20 min. CONCLUSIONS: Intradural en-bloc removal of the ACP with fracture of the optic strut requires minimal drilling, resulting in decreased risk of injury to the optic nerve and a shortened time for clinoidectomy. PMID- 15118889 TI - Intra-suprasellar meningioma mimicking pituitary apoplexy. AB - The clinical and radiological features of an intra-suprasellar meningioma diagnosed as a pituitary macro-adenoma are presented. It is emphasized that this unusual tumour can mimic pituitary apoplexy. Differential diagnosis and surgical treatment of intrasellar meningioma versus pituitary adenoma are discussed with a review of the literature. PMID- 15118890 TI - Severe delayed diffuse cerebral vasospasm and cerebral infarctions following spinal subdural hemorrhage. AB - BACKGROUND: We report a rare case of severe delayed cerebral vasospasm with cerebral infarctions after spinal subdural hemorrhage. CASE REPORT: A 56-year-old woman presented with an acute onset of paraplegia. MR-imaging revealed an extensive intraspinal hemorrhage reaching from T1 to L1. The hematoma was evacuated via a T8-laminectomy. At the 7th postoperative day the patient developed visual disturbances. MR-scanning revealed extensive infarctions and cerebral angiography showed severe diffuse vasospasms. INTERPRETATION: This case demonstrates that cerebral vasospasm may be caused by a spinal subdural hemorrhage, supporting the hypothesis that cerebral vasospasm may be triggered by factors from a remote site and that a direct contact of blood clots with the vessel is not mandatory. PMID- 15118891 TI - Aspergillus pituitary abscess. AB - BACKGROUND: Pituitary abscess is rare and most of the cases are of bacterial origin. True fungal pituitary abscess is extremely rare only five cases have been reported. In this report, we present a case of aspergillus pituitary abscess. Mortality rate in intracranial aspergillosis is close to 100% especially in immunsuppressed patients when undiagnosed and untreated. In focal CNS aspergillosis total cure can be achieved in approximately 30% of the cases by surgical drainage and intensive antifungal therapy. Although this is the first reported case with magnetic resonance imaging examination the definitive diagnosis was established only by histopathological examination. CLINICAL PRESENTATION: A 42 year-old man was referred to our hospital with the diagnosis of sellar suprasellar mass accompanied by frontal headache and decreased visual acuity. His medical history was insignificant. Physical examination was normal and the patient was afebrile. The neurological examination revealed bilateral papilledema and bitemporal hemianopsia but no stiff neck and motor or sensory deficit. In the light of MRI examination, the preoperative diagnosis was pituitary abscess secondary to paranasal sinus infection or hemorrhagic pituitary adenoma. INTERVENTION: The patient was successfully treated by transsphenoidal surgery. Histopathological examination of sphenoid sinus mucosa revealed normal mucosal appearance with inflammation and histopathological examination of the intrasellar mass resulted in the diagnosis of aspergillosis. All cultures obtained from sphenoid sinus were reported as having no growth. However in the second week after the operation fungal culture of the intrasellar mass grew aspergillus. After 8 weeks of amphothericine-B treatment, the patient was discharged. At the last follow up examination two years after the operation, the patient was symptom free with normal pituitary function. CONCLUSION: Aspergillus pituitary abscess should be considered in the differential diagnosis of a pituitary mass. The correct diagnosis of pituitary aspergillosis can only be achieved by histopathological examination because clinical and radiological findings including MRI are not specific and culture results are obtained later. Immediately after the diagnosis, intensive antifungal therapy should be started for a successful treatment. PMID- 15118892 TI - Primary primitive neuro-ectodermal tumor of cauda equina with intracranial seeding. AB - Primitive neuro-ectodermal tumor (PNET) of the cauda equina is a rare entity. 18 cases have been reported in the literature so far, including 4 cases with intracranial seeding. Moreover parenchymal involvement of brain has never been reported as a form of intracranial seeding from PNET of the cauda equina. A 31 year-old female patient, with PNET of cauda equina showing intracranial seeding 6 months after surgery, is presented in this report. To our knowledge, this is the first adult case of spinal cord PNET with parenchymal involvement of brain. The histopathological, clinical and radiological findings as well as treatment of the patient were evaluated. PMID- 15118893 TI - Chronic subdural collection after endoscopic third ventriculostomy. AB - Endoscopic third ventriculostomy (ETV) is considered a safe technique for the treatment of obstructive hydrocephalus. We describe a case of chronic subdural haematoma (CSDH) after ETV, revealed by MRI four weeks after the procedure, and requiring surgical evacuation, in a 69 y.o. asymptomatic male patient. In our opinion, overdrainage may evolve also in endoscopic treatment of obstructive hydrocephalus. This complication could be the starting point of the subdural collection. We review the literature and discuss the causes that may lead to CSDH after ETV procedure. PMID- 15118894 TI - Brain abscess following cerebral infarction. PMID- 15118895 TI - Clivus lymphoma. AB - Lymphoma as primary infiltration of the skull base, especially the clivus, is unusual and rare. However, increasing incidence of atypical lymphoma manifestations have been reported. As cranial base surgery becomes more common, it is important to be aware of lymphoma as a possible differential diagnosis of clivus lesions. This case presents a primary malignant lymphoma of the clivus, histologically confirmed after intra-operative MRI-guided transnasal transsphenoidal biopsy. PMID- 15118896 TI - Post traumatic brain contusions and lacerations. An international focus meeting, September 19-20, 2003, Rimini, Italy. PMID- 15118897 TI - The value of combining single photon emission computerised tomography and computerised tomography in the investigation of spondylolysis. AB - The aim of this study was to assess the diagnostic value of combining single photon emission computerised tomography (SPECT) with reverse gantry computerised tomography (rg-CT) in the investigation of spondylolysis. Patient characteristics and imaging results in 118 patients, aged 8-44 years, with low back pain (LBP) were analysed. SPECT showed increased scintigraphic uptake in 80 patients, and spondylolysis was identified on rg-CT in 53. The Cohen Kappa ratio of 0.362 (95% CI: 0.198-0.526) suggests only fair agreement for the result of increased scintigraphic activity with the finding of spondylolysis on rg-CT. We conclude that these investigations give mutually exclusive information, which leads to four diagnostic categories. When there was increased scintigraphic activity on SPECT, 58.8% (95% CI: 48.0-69.5%) of patients had spondylolysis on rg-CT. With rest from provoking activities, these lesions may heal. We interpret the findings of increased scintigraphic activity, but no spondylolysis demonstrated on rg-CT as indicating a bone stress response. These also require rest from provoking activity to prevent a stress fracture developing. In this study, 84.2% (95% CI: 72.67-95.8%) of those patients without increased activity on SPECT had no spondylolysis identified on rg-CT. These patients may need further investigations such as magnetic resonance imaging (MRI) to diagnose pathology, which typically does not involve the posterior elements--but rest from sport may not be so important. There were five patients in our study, without increased scintigraphic activity, but in whom bilateral chronic-appearing (wide separation, smooth sclerotic bone margins) spondylolyses were identified at L5. These all were anticipated from previous plain radiographs or MRI. This group will almost certainly not heal, and if the spondylolyses are the cause of pain these vertebrae will need stabilisation by surgery if physiotherapy fails. PMID- 15118898 TI - Neurosurgeons' management of lumbosacral radicular syndrome evaluated against a clinical guideline. AB - To establish to what extent neurosurgeons subscribe to the lumbosacral radicular syndrome (LRS) guideline, and to evaluate their current management of patients with LRS against the guideline. All active neurosurgeons in the Netherlands (n=92) were mailed a questionnaire about the guideline and data from 66 responders were analysed. Patients were recruited via seven of the participating neurosurgeons and were interviewed once by telephone. The medical records of the participating patients (n=163) were also examined. Of the 26 propositions in the LRS guideline, seven were not fully endorsed by the neurosurgeons. Three of these seven propositions may need updating based on "new evidence". The time between the onset of the LRS episode and the actual moment of surgery was considerably longer than that recommended in the guideline. Based on their current management of LRS patients, the neurosurgeons largely adhere with the LRS guideline. PMID- 15118899 TI - Impact of cancer patients' quality of life on that of spouse caregivers. AB - GOALS OF WORK: This study aimed to examine the correlation between quality of life (QOL) in cancer patients and that of their spouse caregivers and to identify factors that influence this correlation. PATIENTS AND METHODS: This cross sectional study collected data from 121 cancer patient/spouse caregiver dyads. The Functional Assessment of Cancer Therapy Scale-General (FACT-G) was used to measure patients' QOL, and the Caregiver Quality of Life Index (CQLI) was used to measure spouse caregivers' QOL. Correlation coefficients between patients' and caregivers' QOL were computed for four dimensions of QOL, as well as a total score for QOL. Correlations between patients' and caregivers' total QOL scores were furthered analyzed by three groups of factors: disease-/treatment-related, caregiving-related, and relationship-related variables. MAIN RESULTS: Only the social/family and functional dimensions of patient QOL and total score for patient QOL were associated with each dimension of their caregivers' QOL and with the total score ( r=0.27-0.44). Physical and emotional dimensions of patients' QOL did not significantly influence spouse caregivers' QOL for any dimension nor for the total score. Factors influencing the association between patients' and caregivers' overall QOL included cancer diagnosis, length of hospitalization, caregiving intensity and duration, marital satisfaction, and caregiving self esteem. CONCLUSIONS: Social and functional aspects of patients' QOL play a significant role in determining the QOL of their spouse caregivers. The strength of association between patients' and spouse caregivers' overall QOL can be moderated by some factors. PMID- 15118900 TI - Egg size and offspring performance in the collared flycatcher ( Ficedula albicollis): a within-clutch approach. AB - Adaptive within-clutch allocation of resources by laying females is an important focus of evolutionary studies. However, the critical assumption of these studies, namely that within-clutch egg-size deviations affect offspring performance, has been properly tested only rarely. In this study, we investigated effects of within-clutch deviations in egg size on nestling survival, weight, fledgling condition, structural size and offspring recruitment to the breeding population in the collared flycatcher ( Ficedula albicollis). Besides egg-size effects, we also followed effects of hatching asynchrony, laying sequence, offspring sex and paternity. There was no influence of egg size on nestling survival, tarsus length, condition or recruitment. Initially significant effect on nestling mass disappeared as nestlings approached fledging. Thus, there seems to be limited potential for a laying female to exploit within-clutch egg-size variation adaptively in the collared flycatcher, which agrees with the majority of earlier studies on other bird species. Instead, we suggest that within-clutch egg-size variation originates from the effects of proximate constraints on laying females. If true, adaptive explanations for within-clutch patterns in egg size should be invoked with caution. PMID- 15118901 TI - Effects of elevated atmospheric CO2 on the nutritional ecology of C3 and C4 grass feeding caterpillars. AB - It is plausible that the nutritional quality of C3 plants will decline more under elevated atmospheric CO2 than will the nutritional quality of C4 plants, causing herbivorous insects to increase their feeding on C3 plants relative to C4 plants. We tested this hypothesis with a C3 and C4 grass and two caterpillar species with different diet breadths. Lolium multiflorum (C3) and Bouteloua curtipendula (C4) were grown in outdoor open top chambers at ambient (370 ppm) or elevated (740 ppm) CO2. Bioassays compared the performance and digestive efficiencies of Pseudaletia unipuncta (a grass-specialist noctuid) and Spodoptera frugiperda (a generalist noctuid). As expected, the nutritional quality of L. multiflorum changed to a greater extent than did that of B. curtipendula when grown in elevated CO2; levels of protein (considered growth limiting) declined in the C3 grass, while levels of carbohydrates (sugar, starch and fructan) increased. However, neither insect species increased its feeding rate on the C3 grass to compensate for its lower nutritional quality when grown in an elevated CO2 atmosphere. Consumption rates of P. unipuncta and S. frugiperda were higher on the C3 grass than the C4 grass, the opposite of the result expected for a compensatory response to the lower nutritional quality of the C4 grass. Although our results do not support the hypothesis that grass-specialist insects compensate for lower nutritional quality by increasing their consumption rates more than do generalist insects, the performance of the specialist was greater than that of the generalist on each grass species and at both CO2 levels. Mechanisms other than compensatory feeding, such as increased nutrient assimilation efficiency, appear to determine the relative performance of these herbivores. Our results also provide further evidence against the hypothesis that C4 grasses would be avoided by insect herbivores because a large fraction of their nutrients is unavailable to herbivores. Instead, our results are consistent with the hypothesis that C4 grasses are poorer host plants primarily because of their lower nutrient levels, higher fiber levels, and greater toughness. PMID- 15118902 TI - A critical evaluation of intrapopulation variation of delta13C and isotopic evidence of individual specialization. AB - Individual variation in the diet of consumers is common in many ecological systems and has important implications for the study of population dynamics, animal behavior, and evolutionary or ecological interactions. Ecologists frequently quantify the niche of a population by intensive analyses of gut contents and feeding behaviors of consumers. Inter-individual differences in delta13C signature can indicate long term differences in feeding behavior, often unattainable by a single snapshot analysis of gut contents. If a consumer's food sources have unique delta13C signatures, then the intrapopulation variation in delta13C may be useful for quantifying diet variation and detecting isotopic evidence of individual specialization. However, intrapopulation variation in delta13C can underestimate or overestimate dietary variation, and therefore is not directly equivalent to a dietary based niche. In this paper we show that intrapopulation variability of delta13C in consumers critically depends on the isotopic range and distribution of food sources. Our analyses fundamentally challenge how we interpret the intrapopulation isotopic variance of delta13C, and how we evaluate isotopic evidence of individual specialization. PMID- 15118903 TI - Expression of DNase gamma during Fas-independent apoptotic DNA fragmentation in rodent hepatocytes. AB - Endonuclease-induced DNA fragmentation is a hallmark of apoptosis. DNase gamma (DNase gamma) was recently identified as one of the endonucleases responsible for apoptotic DNA fragmentation. In this study, immunohistochemistry for DNase gamma was performed on paraffin sections of rodent liver in well-defined models of hepatocyte apoptosis induced by Fas antibody (Fas) or cycloheximide (CHX), and necrosis induced by lipopolysaccharide (LPS) or carbon tetrachloride (CCl4). DNase gamma immunoreactivity was compared with TdT-mediated dUTP nick-end labeling (TUNEL) reactivity. Our results showed TUNEL reactivity in both apoptotic and necrotic hepatocytes. DNase gamma immunoreactivity was not detected during LPS-induced or CCl4-induced hepatocyte necrosis. In contrast, it was evident during CHX-induced, but not Fas-induced, apoptotic DNA fragmentation. These findings suggest that DNase gamma plays an important role in Fas independent apoptotic DNA fragmentation in hepatocytes. PMID- 15118904 TI - An intragenic deletion/inversion event in the DMD gene determines a novel exon creation and results in a BMD phenotype. AB - Duchenne and Becker Muscular Dystrophy (DMD and BMD) are caused, in the majority of cases, by deletions in the dystrophin gene ( DMD). Here we describe the unprecedented case of a BMD patient carrying a large out-of-frame intragenic deletion, together with an inversion in the DMD gene, resulting in the inclusion of a novel exon in the transcript. Multiplex PCR amplification revealed the presence of a 48-52 exon deletion, but transcript analysis identified two unexpected products, neither of them including exon 53. The shorter mRNA derived from the juxtaposition of exons 47-54 (in-frame), while the longer one resulted from the inclusion of a novel 73-bp exon between exons 47 and 54. Sequence analysis revealed that the inserted sequence derived from an inverted portion of intron 53; its inclusion is predicted to determine protein truncation. The presence of a genomic inversion involving exon 53 and flanking regions was confirmed, and inversion/deletion breakpoints were sequenced. The inverted 73-bp sequence displays splicing signals at both ends and thus it is probably recognized as a novel exon when the partially inverted hnRNA is processed. These findings highlight the importance of mRNA analysis on patients that, based on routine DNA screenings, do not follow the reading-frame rule. This is the first reported patient carrying both an intragenic deletion and inversion in the DMD locus. This case might provide further insight into both the mechanisms that determine genomic rearrangements in the DMD locus and the molecular signals that drive exon inclusion. PMID- 15118905 TI - Ancestral proportions and their association with skin pigmentation and bone mineral density in Puerto Rican women from New York city. AB - Hispanic and African American populations exhibit an increased risk of obesity compared with populations of European origin, a feature that may be related to inherited risk alleles from Native American and West African parental populations. However, a relationship between West African ancestry and obesity related traits, such as body mass index (BMI), fat mass (FM), and fat-free mass (FFM), and with bone mineral density (BMD) in African American women has only recently been reported. In order to evaluate further the influence of ancestry on body composition phenotypes, we studied a Hispanic population with substantial European, West African, and Native American admixture. We ascertained a sample of Puerto Rican women living in New York ( n=64), for whom we measured BMI and body composition variables, such as FM, FFM, percent body fat, and BMD. Additionally, skin pigmentation was measured as the melanin index by reflectance spectroscopy. We genotyped 35 autosomal ancestry informative markers and estimated population and individual ancestral proportions in terms of European, West African, and Native American contributions to this population. The ancestry proportions corresponding to the three parental populations are: 53.3+/-2.8% European, 29.1+/ 2.3% West African, and 17.6+/-2.4% Native American. We detected significant genetic structure in this population with a number of different tests. A highly significant correlation was found between skin pigmentation and individual ancestry ( R(2)=0.597, P<0.001) that was not attributable to differences in socioeconomic status. A significant association was also found between BMD and European admixture ( R(2)=0.065, P=0.042), but no such correlation was evident with BMI or the remaining body composition measurements. We discuss the implications of our findings for the potential use of this Hispanic population for admixture mapping. PMID- 15118906 TI - The mitochondrial IMP peptidase of yeast: functional analysis of domains and identification of Gut2 as a new natural substrate. AB - The mitochondrial inner membrane peptidase IMP of Saccharomyces cerevisiae is required for proteolytic processing of certain mitochondrially and nucleus encoded proteins during their export from the matrix into the inner membrane or the intermembrane space. The membrane-associated signal peptidase complex is composed of the two catalytic subunits, Imp1 and Imp2, and the Som1 protein. The IMP subunits are thought to function in membrane association, interaction and stabilisation of subunits, substrate specificity, and proteolysis. We have analysed inner membrane peptidase mutants and substrates to gain more insight into the functions of various domains and investigate the basis of substrate recognition. The results suggest that certain conserved glycine residues in the second and third conserved regions of Imp1 and Imp2 are important for stabilisation of the Imp complex and for the proteolytic activity of the subunits, respectively. The non-conserved C-terminal parts of the Imp subunits are important for their proteolytic activities. The C-terminal region of Imp2, comprising a predicted second transmembrane segment, is dispensable for the stability of Imp2 and Imp1, and cannot functionally substitute for the C-terminal segment of Imp1. Alteration of the Imp2 cleavage site in cytochrome c(1) (from A/M to N/D) reveals the specificity of the Imp2 peptidase. In addition, we have identified Gut2, the mitochondrial FAD-dependent glycerol-3-phosphate dehydrogenase, as a new substrate for Imp1. Failure to cleave the Gut2 precursor may contribute to the pet phenotype of certain imp mutants. Gut2 is associated with the inner membrane, and is essential for growth on glycerol-containing medium. Suggested functions of the analysed residues and domains of the IMP subunits, characteristics of the cleavage sites of substrates and implications for the phenotypes of imp mutants are discussed. PMID- 15118907 TI - Navigation within host tissues: cercariae orientate towards dark after penetration. AB - The survival of skin penetrating cercariae depends on information on the direction to move toward deeper layers in the epidermis (the direction of further migration) and toward the surface (direction which must be avoided when migrating). We tested the hypothesis that parasites can use their photo sensitivity for orientation away from the light-exposed skin surface towards darker locations. Cercariae of species invading humans ( Schistosoma mansoni), birds ( Trichobilharzia ocellata) and fish ( Diplostomum spathaceum) oriented towards light sources when free swimming in cuvettes. However, they shifted to a negative photo-orientation when migrating in agar substrates after penetration and transformation to schistosomula. This is a first hint that parasites may use photo-orientation when they navigate in host tissues. PMID- 15118908 TI - Ultrastructural particularities of the spermatozoon of the cestode Electrotaenia malopteruri (Fritsch, 1886) (Proteocephalidae: Gangesiinae), a parasite of Malapterurus electricus (Siluriformes: Malapteruridae) from the river Nile, Egypt. AB - The fine structure of the mature spermatozoon of the tapeworm Electrotaenia malopteruri (Fritsch, 1886), a member of a supposedly primitive group of proteocephalidean tapeworms (Proteocephalidae: Gangesiinae), was studied by transmission electron microscopy for the first time. The mature spermatozoon of E. malopteruri is filiform, tapers at both extremities, and contains two axonemes of 9+"1" trepaxonematan type. A helicoidal crested body (60-150 nm thick) is present at the anterior extremity of the gamete. The twisting cortical microtubules (CM) line the periphery of the spermatozoon continuously. The nucleus, a fine cord of moderately condensed chromatin, occupies the middle part (region III) of the spermatozoon. The slightly electron-dense cytoplasm contains electron-dense granules in regions II and III and becomes more electron-dense at the end of region IV. The anterior and posterior extremities of the spermatozoon contain a single axoneme. Two modes of disorganisation of the axoneme at the posterior end of the mature spermatozoa are described for the first time in cestodes. The present data also indicate that the sperm ultrastructure of E. malapteruri shows some characters typical for onchobothriid tetraphyllideans, but it resembles that of Cyclophyllidea in the arrangement of twisting of the CM. PMID- 15118909 TI - Ultrastructure of spermiogenesis and the spermatozoon of Monorchis parvus Looss, 1902 (Digenea, Monorchiidae), a parasite of Diplodus annularis (Pisces, Teleostei). AB - The present paper describes the characteristic ultrastructural features of spermiogenesis and the spermatozoon of Monorchis parvus (Trematoda, Digenea, Monorchiidae). This spermiogenesis is characterized by a flagellar rotation of about 120 degrees and the formation of a novel ultrastructural element: a centriolar extension. It nevertheless follows the general pattern of digeneans. It begins with the formation of a differentiation zone, comprising striated rootlets associated with two centrioles and an intercentriolar body. The mature spermatozoon presents features allowing the distinction between M. parvus and other digenetic trematodes. It possesses an original anterior extremity. The two central elements of the axonemes appear prior to the peripheral doublets. The external ornamentations of the cell membrane are formed parallel to the central elements. Another peculiarity of the male gamete is the presence of two mitochondria. M. parvus, therefore, differs distinctly from other digenetic trematodes. PMID- 15118910 TI - Serotype distribution and antimicrobial susceptibility of Streptococcus pneumoniae causing invasive infections and acute otitis media in children. AB - A prospective study was conducted to determine the serotypes and antibiotic resistance patterns of pneumococcal isolates from children with invasive pneumococcal disease (IPD) and acute otitis media (AOM). From October 2001 to May 2002, 65 children with IPD (28 bacteraemic pneumonia, 24 bacteraemia without focus, 7 meningitis, 6 other infections) and 78 with AOM were identified. The most common serotypes causing IPD were 14 (32.3%), 6B (20.0%), 1 (18.5%) and 19F (7.7%) whereas the predominant serotypes causing AOM were 19F (35.9%), 14 (16.7%) and 23F (9.1%). Sixty-nine percent of IPD and 70.5% of AOM were caused by vaccine serotypes. The vaccine serotypes were more commonly encountered in meningitis cases and in children younger than 2 years of age. Intermediate resistance to penicillin was observed in 6 of 65 (9.2%) IPD isolates, one of which was intermediately resistant to cefotaxime (1.6%), whereas none exhibited high-level resistance to penicillin or other beta-lactam antibiotics. A higher proportion of antimicrobial resistance was noted in AOM isolates; 29 of 78 (37.4%) exhibited intermediate resistance and 8 (10.2%) high level resistance to penicillin, four of which had intermediate resistance to cefotaxime. Significant resistance was also noted to erythromycin; 38.5% of IPD and 48.7% of AOM isolates were resistant. Multidrug resistance was observed in one IPD and in eight AOM isolates. CONCLUSION: these findings have implications in the potential use of 7 valent conjugate vaccine in our region. PMID- 15118911 TI - Stability and inactivation of SARS coronavirus. AB - The SARS-coronavirus (SARS-CoV) is a newly emerged, highly pathogenic agent that caused over 8,000 human infections with nearly 800 deaths between November 2002 and September 2003. While direct person-to-person transmission via respiratory droplets accounted for most cases, other modes have not been ruled out. Faecal shedding is common and prolonged and has caused an outbreak in Hong Kong. We studied the stability of SARS-CoV under different conditions, both in suspension and dried on surfaces, in comparison with other human-pathogenic viruses, including human coronavirus HCoV-229E. In suspension, HCoV-229E gradually lost its infectivity completely while SARS-CoV retained its infectivity for up to 9 days; in the dried state, survival times were 24 h versus 6 days. Thermal inactivation at 56 degrees C was highly effective in the absence of protein, reducing the virus titre to below detectability; however, the addition of 20% protein exerted a protective effect resulting in residual infectivity. If protein containing solutions are to be inactivated, heat treatment at 60 degrees C for at least 30 min must be used. Different fixation procedures, e.g. for the preparation of immunofluorescence slides, as well as chemical means of virus inactivation commonly used in hospital and laboratory settings were generally found to be effective. Our investigations confirm that it is possible to care for SARS patients and to conduct laboratory scientific studies on SARS-CoV safely. Nevertheless, the agents tenacity is considerably higher than that of HCoV-229E, and should SARS re-emerge, increased efforts need to be devoted to questions of environmental hygiene. PMID- 15118912 TI - Histological organization of roe deer testis throughout the seasonal cycle: variable and constant components of tubular and interstitial compartment. AB - Seasonally regulated breeding in roe deer, Capreolus capreolus, is associated with significant changes in testis mass, structure and function. This study has quantified seasonal changes of morphometric parameters and cellular composition in roe deer testis parenchyma. Tissue samples were collected bimonthly during a complete annual cycle. Morphometric parameters of seminiferous tubules were measured and the number of different cell types was counted using a computer aided image-analyzing system. A scheme of eight tubular epithelium stages for active spermatogenesis was devised according to the spermatid development. Stage I is characterized by the occurrence of new round spermatids, stage IV by spermiation and stage VIII by the meiotic division of spermatocytes. The average diameter of seminiferous tubules varied between 88.4+/-3.6 micro m (February) and 216.8+/-9.2 micro m (June). Also numbers of spermatogonia, spermatocytes and spermatids per tubule cross-section showed considerable seasonal changes. In December and February the germinative epithelium mainly consists of Sertoli cells and spermatogonia. In February, the first differentiated spermatogonia enter meiosis, and in April even spermatids occasionally occur, which reach their highest numbers during the rut in August. Both the expansion and the proportion of tubular and interstitial compartment change seasonally and result in differing cell densities. Assuming numerically constant populations of Sertoli cells and interstitial cells during the entire year, the hypothetical cell numbers per mm(2) of the tubular and interstitial areas were calculated for the seasonally variable total areas of tissue cross-sections. The concordance of these theoretical values with measured cell densities provided evidence that the total numbers of Sertoli cells, as well as interstitial cells, remain really constant throughout the seasonal cycle. The exact quantification of variable and constant components provides basic data for characterization of cell type and stage specific processes of spermatogenesis. PMID- 15118913 TI - Postnatal development of the reproductive system in the grey short-tailed opossum, Monodelphis domestica. AB - Postnatal phenotypic sex differentiation has been investigated in a laboratory marsupial, Monodelphis domestica, as part of a larger study to resolve apparent discrepancies between eutherian and marsupial mammals. These include the formation of sex-specific structures in marsupials prior to gonadal differentiation and the retention in both sexes of structures which are sex specific in eutherians. The time-course and nature of differentiation was investigated in 131 specimens ranging in age from the day of birth to 56 days. Patent wolffian ducts extend to the urogenital sinus in both sexes at birth, while mullerian ducts are identified on day 1 and grow in a cranio-caudal direction to reach the urogenital sinus on day 6. The male mullerian duct shows signs of regression at its cranial end on day 10 and throughout its length on day 12; its lumen has completely disappeared by day 15. By this time the epididymis and vas deferens have developed from the wolffian duct; their histological differentiation occurs between days 26 and 56. Prostatic buds are identifiable in tissue surrounding the male urethra on day 14. In the female, the wolffian duct is larger than the mullerian duct until day 14; thereafter the wolffian duct begins to regress at its cranial end, disappearing by day 17, whereas the mullerian duct begins to enlarge, converging with its fellow at the urogenital sinus by day 19. Lateral vaginae, vaginal culs-de-sac, uteri and oviducts have differentiated from the mullerian ducts by day 25. Gonads of both sexes are elongated in shape at birth, attached along the medial aspect of the large mesonephroi in the abdominal cavity. However, from day 3 onwards the testis becomes more rounded than the ovary. Degeneration of the male mesonephros begins about day 10 and is almost completed by day 19; the female mesonephros is still relatively large at day 14 though it too has almost disappeared by day 19. By postnatal day 13 the abdominal phase of testis descent is underway and the inguinal phase begins at day 15. Testes have reached the scrotal sac by day 24 and achieve their final position at the base of the scrotum by day 28. In summary, postnatal reproductive tract development and gonadal descent has been examined in this important biomedical model, where differentiation of the wolffian and mullerian ducts takes place after gonadal differentiation, according to the normal eutherian pattern. PMID- 15118914 TI - Looking through genomics: concepts and technologies for plant and animal genomics. PMID- 15118915 TI - One-encounter search-image formation by araneophagic spiders. AB - An experimental study of search-image use by araneophagic jumping spiders (i.e., salticid spiders that prey routinely on other spiders) supports five conclusions. First, araneophagic salticids have an innate predisposition to form search images for specific prey from their preferred prey category (spiders) rather than for prey from a non-preferred category (insects). Second, single encounters are sufficient for forming search images. Third, search images are based on selective attention specifically to optical cues. Fourth, there are trade-offs in attention during search-image use (i.e., forming a search image for one type of spider diminishes the araneophagic salticid's attention to other spiders). Fifth, the araneophagic salticid's adoption of search images is costly to the prey (i.e., when the araneophagic salticid adopts a search, the prey's prospects for surviving encounters with the araneophagic salticid are diminished). Cognitive and ecological implications of search-image use are discussed. PMID- 15118916 TI - Up-regulation of transcriptional factor E2F1 in papillary and anaplastic thyroid cancers. AB - Expression of genes in the Rb-E2F signaling pathway is controlled by E2F transcriptional factors originally defined as molecules that bind to the promoter of E2 adenovirus. The E2F gene family consists of six members and is designated E2F1-6. The Rb-E2F signaling pathway is among the main regulators of the cell cycle, hence its importance in differentiation and oncogenesis. We document here up-regulation of E2F1, but not other members of the E2F gene family, in 15 of 18 primary papillary thyroid cancers examined (83%) in comparison to corresponding noncancerous thyroid tissues and in all of 11 anaplastic thyroid cancer (ATC) cell lines (100%). The E2F4 gene, however, was down-regulated in 12 of the papillary thyroid cancers (67%). Immunohistochemical analysis with antibody to E2F1 revealed prominent intracellular E2F1 protein in most of the primary papillary cancers (16 of 18; 89%) but was not detectable in normal thyroid tissues. These data indicated that increased expression of the E2F1 gene might play a significant role in human thyroid carcinogenesis through derangement of the Rb-E2F signaling pathway. PMID- 15118917 TI - [Predictors of outcome in patients with normal pressure hydrocephalus]. AB - Despite emerging knowledge over 40 years, the postoperative results after shunt implantations in patients diagnosed for normal pressure hydrocephalus (NPH) did not improve significantly during the last decade. For that reason predictors have to be identified in order to predict preoperatively the course of disease. From 1982 until 2000 we examined in a prospective study 200 patients diagnosed for NPH. From the patients, who were surgically treated by a shunt implantation we could reexamine 155 (78%) in a meantime interval of seven month after the operation. The NPH was graduated according to the results of the intrathecal infusion test in an early state NPH (without brain atrophy) and late state NPH (with brain atrophy). In our study, we focussed the attention on the possible predictors: patients age, length of disease, clinical signs--like gait ataxia, dementia and bladder incontinence, aetiology idiopathic/secondary as well as implanted valve type and the value of resistance to cerebrospinal fluid outflow. To measure the outcome we used the NPH-Recovery-Rate, as statistical test the Chi square according to Pearson. In 80 patients with an early stage NPH (without cerebral atrophy) a short course of disease (< 1 year), a just slight distinct of dementia and an implanted Miethke-Dual-Switch valve were significant predictors for a positive postoperative outcome. The outflow resistance measured in the intrathecal infusion test showed only a minimal relevance for the outcome. Those 75 patients with a late state NPH (with cerebral atrophy) had a better outcome when dementia was not present, the outflow resistance was above 20 mm Hg*min/ml, the CSF tap-test was positive and a Miethke-Dual-Switch valve was implanted. PMID- 15118918 TI - Immediate postoperative angiography after aneurysm clipping--implications for quality control and guidance of further management. AB - BACKGROUND: To determine the impact of postoperative angiography after aneurysm clipping on quality control and further management. METHODS: A recent consecutive series of n = 296 patients (186 females, 110 males, mean age 51 yrs) who underwent 324 craniotomies to clip 384 aneurysms was reviewed. New irregularities on postoperative angiographies were categorized as unrelated (e. g. vasospasm) or related (= vessel occlusion/residuum) to clipping and expected or unexpected by the surgeon. Therapies prompted by these findings as well as subsequent negative sequelae were analyzed. Factors related to negative events were identified by logistic regression (p < 0.05). RESULTS: Twenty-six (8%) unrelated findings prompted medical or intravascular therapy. Of 36 (9.4%) clip-related findings (n = 17 occlusions, n = 19 residua), 14 (3.6%) were unexpected (n = 9 occlusions, n = 5 residua). This was followed by 9 (2.3%, n = 4 occlusions, n = 5 residua) clip readjustments and 6 aggressive medical therapies. Nine patients were scheduled for angiographic follow-up, of which in five performed so far no change of aneurysm remnants was noted. Permanent sequelae from vessel occlusion occurred in 9 cases. Major premature rupture (p < 0.005), giant size/fusiform configuration (p < 0.001), posterior circulation aneurysms (p < 0.05) and ophthalmic segment location (p < 0.008) were significantly related to adverse postoperative findings. The use of microvascular Doppler significantly reduced the rate of unexpected vessel occlusion (3.3 vs. 0.9%). CONCLUSIONS: Apart from the need for quality control in aneurysm clipping--especially with respect to coiling--our results stress the importance of postoperative angiography, because a small, but significant subset of patients will benefit from immediate intervention and/or deserves long-term follow-up. PMID- 15118919 TI - Clinical application of neuro-navigation in a series of single burr-hole procedures. AB - With recent developments in computer technology and the improvement of neuroimaging, modern optical neuro-navigation systems are increasingly being used in neurosurgery. In this study, we present our experience with 51 operations using a frameless optical navigation system in a variety of single burr-hole procedures. The procedures include neuroendoscopic surgery, frameless stereotactic biopsy, cyst aspiration and catheter placement. Both the VectorVision and the VectorVision(2) neuro-navigation systems (BrainLab AG, Munich, Germany) were used. The reliability and accuracy of the neuro-navigation system, postoperative complications and the clinical usefulness of image-guidance were analyzed. The navigation system worked properly in all 51 neurosurgical cases. Exact planning of the approach and determination of the ideal trajectory were possible in all cases. The mean registration error of the system, given as a computer-calculated value, was 2.1 mm (0.4-3.1 mm). Postoperative clinical evaluations and imaging were performed on every patient in order to confirm the success of the surgical procedure. All patients recovered well and without any postoperative complications. We conclude that image guidance in single burr-hole procedures provides a high degree of accuracy in lesion targeting, permits good anatomical orientation and minimizes brain trauma. The navigation system has proven to be a helpful tool since it increases the safety of single burr-hole procedures. PMID- 15118920 TI - Technical aspects and results of surgery for craniosynostosis. AB - BACKGROUND: The timing and extent of surgery continue to be a matter of discussion in current craniosynostosis management. PATIENTS AND METHODS: We retrospectively reviewed the first 159 children seen from 1996 to 2000 of a total of 243 patients presenting with suspected craniosynostosis. We did not establish a surgical indication in 74 children (47%), either because their malformations were of different origin without associated craniosynostosis (11/7%) or they presented with only mild clinical manifestations of sutural synostosis (63/40%). In 85 cases (53%), we established a surgical indication. Parents did not follow our recommendation in two cases (1%). In 83 cases, we performed diagnosis-related remodeling at the age of 4-12 months. Demographic data, clinical follow-up findings, and regular photo documentation were analyzed. RESULTS: After a mean follow-up of 3 years and 8 months, the results were satisfactory in 68 cases (82%). Of 15 cases (18%) with only limited improvement, two children were submitted to a second intervention. Another two children had to be reoperated on immediately for insufficient hemostasis. In the largest subgroup of children (47) with sagittal synostosis, there was no difference in outcome between surgery performed before and after 6 months of age. DISCUSSION: Surgical indication and assessment of results are based on subjective criteria which limit the comparison with the literature. Our results correspond to those of similar published series. Most of the unfavorable results could be attributed either to inadequate adaptation of the surgical technique to the individual form of the malformation or to a significant secondary loss of correction in multi-sutural synostosis. CONCLUSIONS: The overall results of surgery within the first year of life were satisfactory in 82%. Standard procedures need to be adapted carefully to the individual form of craniosynostosis to avoid unfavorable results. We therefore propose a new surgical technique based on a statistically averaged modular system of skull models to replace freehand remodeling. PMID- 15118921 TI - Planar brain surface reformations for localization of cortical brain lesions. AB - Cortical lesions close to the central region may be difficult to localize on orthogonal MRI slices, especially when space-occupying lesions distort the brain surface relief and obscure anatomical landmarks. The purpose of this study was to evaluate, whether localization can be improved with planar brain surface (Mercator or "pancake") reformations. Three independent MRI readers localized superficial brain lesions of 30 patients on orthogonal MRI slices and on planar brain surface views reformatted from a sagittal 3D T(1)-weighted gradient echo sequence. On orthogonal MRI slices 46% of lesions were considered easy to localize, 37% difficult and 17% could not be localized. Corresponding values for planar brain surface views were 82%, 16% and 2%, respectively. Evaluation of orthogonal MRI slices took 190 min compared to 100 min for planar brain surface views. Thus, planar brain surface reformations are a helpful and time-saving means to localize superficial brain lesions. PMID- 15118922 TI - Lipofundin-induced intracranial pressure rise after severe traumatic brain injury -a case report. AB - Early nutrition is a recognized component of neurosurgical intensive care treatment. The authors present the case of a patient suffering from severe traumatic brain injury who responded with reproducible intracranial pressure (ICP) crisis to infusion of Lipofundin, a fatty soybean oil-based emulsion for parenteral nutrition. During the described ICP rise, the patient remained hemodynamically stable, therefore an anaphylactic reaction seems to be unlikely. An increase of brain tissue oxygenation parallel to the ICP rise in this case is suggestive for increased cerebral blood flow as a cause of ICP elevation after application of Lipofundin. Without multimodal monitoring and data storage, the described side effect of Lipofundin in our patient would have been difficult to identify. PMID- 15118923 TI - Adult eosinophilic granuloma of the lumbar spine with atypical dissemination. Case report: a long-term follow-up. AB - Vertebral Eosinophilic Granuloma (EG) is a frequently reported disease, but further dissemination rarely occurs when the initial lesion is in the lumbar region. We present a case of EG, initially located in L5 vertebra, which disseminated to the skeletal bones and pulmonary system months after total excision in an adult. PMID- 15118924 TI - Lipomas of the internal auditory canal--report of two cases and review of the literature. AB - OBJECTIVE: Lipomas of the internal auditory canal are extremely rare. So far, only 21 cases have been described in the literature. We present here two of our own cases and review the literature to date with special emphasis on the diagnostic and therapeutic options. METHOD: During the last seven years two lipomas of the internal auditory canal were surgically treated in our department. Despite native and gadolinium-enhanced MRI and a thin-sliced temporal bone CT scan they were misdiagnosed as intracanalicular acoustic neurinomas. RESULTS: Total tumour removal could be achieved, the facial nerve function was conserved, but both patients were rendered functionally deaf after surgery. CONCLUSION: Lipomas' radiological behaviour may mimic acoustic neurinomas; without fat suppressed T(1)-weighted images they are often preoperatively misdiagnosed as acoustic neurinomas. Although the postoperative morbidity with respect to cochlear and facial nerve function is much higher than in small acoustic schwannomas, early surgery may be justified, because complete removal is only possible in this stage. No reliable data are available concerning the natural history of this kind of lesion. PMID- 15118925 TI - Commentary on the Article of C. Woertgen, O. W. Ullrich, R. D. Rothoerl, A. Brawanski: Comparison of the Claassen and Fisher CT classification scale to predict ischemia after aneurysmatic SAH. Zentralbl Neurochir 2003; 64: 104-108. PMID- 15118928 TI - Preclinical gene therapy studies for hemophilia using adeno-associated virus (AAV) vectors. AB - Gene therapy offers a potential cure for hemophilia and several gene transfer vectors have been evaluated for their ability to treat this disease. This article reviews the studies that have been performed to evaluate the ability of recombinant adeno-associated virus (AAV) vectors to achieve safely the sustained expression of clotting factors following intramuscular, intravenous, and intrahepatic delivery to several animal models. These routes of administration are all effective in providing sustained and therapeutic levels of factor IX (FIX), although the levels vary. Intrahepatic delivery is more efficacious than intravenous administration, which is superior to intramuscular delivery. The recent development of efficient factor VIII (FVIII) expression cassettes has made AAV-based gene therapy for hemophilia A also within reach. Although no acute toxicity has been observed with any route of administration, an increased risk of antibody formation against FIX has been noted following intramuscular delivery. Biodistribution studies concluded that the vector disseminates to most tissues in a dose-dependent and time-dependent manner, but the majority of the vector resides in the targeted tissue. In addition, the risk of germline transmission has been shown to be low or absent. The relatively recent isolation of new AAV serotypes has resulted in the identification of vectors that have enhanced tropism for certain tissues. This combined with the potential of these new vectors to evade the immune response to AAV2, makes them attractive candidates for gene therapy. Although much progress has been made using AAV to treat hemophilia, there are several outstanding issues that need to be addressed. Delivery of AAV to large animals has not been reproducible, which could be due to nonoptimized delivery and/or immune responses to the vector or transgene product. In addition, a complete understanding of the biology of these vectors is required to assess their long-term safety. Solving these issues will lead to the development of a successful gene therapy product. PMID- 15118929 TI - Preclinical gene therapy studies for hemophilia using adenoviral vectors. AB - Hemophilia A and B are hereditary coagulation defects resulting from a deficiency of factor VIII (FVIII) and factor IX (FIX), respectively. Introducing a functional FVIII or FIX gene could potentially provide a cure for these bleeding disorders. Adenoviral vectors have been used as tools to introduce potentially therapeutic genes into mammalian cells and are by far the most efficient vectors for hepatic gene delivery. Long-term expression of both FVIII and FIX has been achieved in preclinical (hemophilic) mouse models using adenoviral vectors. Therapeutic levels of FVIII and FIX also have been achieved in hemophilic dogs using adenoviral vectors and in some cases expression was long-term. The performance of earlier generation adenoviral vectors, which retained residual viral genes, was compromised by potent acute and chronic inflammatory responses that contributed to significant toxicity and morbidity and short-term expression of FVIII and FIX. The development of improved adenoviral vectors devoid of viral genes (gutless or high-capacity adenoviral vectors) was therefore warranted, which led to a significant reduction in acute and chronic toxicity and more prolonged expression of FVIII and FIX. Strategies aimed at making these vectors safer and less immunogenic and their implications for hemophilia gene therapy are discussed in this review. PMID- 15118930 TI - Onco-retroviral and lentiviral vector-based gene therapy for hemophilia: preclinical studies. AB - Hemophilia A and B gene therapy requires long-term and stable expression of coagulation factor VIII (FVIII) or factor IX (FIX), respectively, and would need to compare favorably with protein replacement therapy. Onco-retroviral and lentiviral vectors are attractive vectors for gene therapy of hemophilia. These vectors have the potential for long-term expression because they integrate stably in the target cell genome. Whereas onco-retroviral vectors can only transduce dividing cells, lentiviral vectors can transduce a broad variety of cell types irrespective of cell division. Several preclinical and clinical studies have explored the use of onco-retroviral and, more recently, lentiviral vectors for gene therapy of hemophilia A or B. Both ex vivo and in vivo gene therapy approaches have been evaluated, resulting in therapeutic FVIII or FIX levels in preclinical animal models. Whereas in vivo gene therapy using onco-retroviral or lentiviral vectors often led to long-term FVIII or FIX expression from transduced hepatocytes, ex vivo approaches were generally hampered by either low or transient expression of FVIII or FIX levels in vivo and/or inefficient engraftment. Furthermore, immune responses against the transgene product remain a major issue that must be resolved before the full potential of these vectors eventually can be exploited clinically. Nevertheless, the continued progress in vector design combined with a better understanding of vector biology may ultimately yield more effective gene therapy approaches using these integrating vectors. PMID- 15118931 TI - Nonviral gene therapy approaches to hemophilia. AB - The goal of hemophilia gene therapy is to obtain long-term therapeutic levels of factor VIII (FVIII) or factor IX (FIX) without stimulating an immune response against the transgene product or the vector. The success of gene therapy is largely dependent on the development of appropriate gene delivery vectors. Both viral vectors and nonviral vectors have been considered for the development of hemophilia gene therapy. In general, viral vectors are far more efficient than nonviral gene delivery approaches and resulted in long-term therapeutic levels of FVIII or FIX in preclinical animal models. However, there are several reasons why a nonviral treatment would still be desirable, particularly because some viral vectors are associated with inflammatory reactions, that render transgene expression transient, or with an increased risk of insertional oncogenesis when random integrating vectors are used. Nonviral vectors may obviate some of these concerns. Since nonviral vectors are typically assembled in cell-free systems from well-defined components, they have significant manufacturing advantages over viral vectors. The continued development of improved nonviral gene delivery approaches offers new perspectives for gene therapy of chronic diseases including hemophilia. PMID- 15118932 TI - Preclinical animal models for hemophilia gene therapy: predictive value and limitations. AB - Hemophilia A and B are excellent candidate disorders for the application of somatic cell gene therapy. One of the major advantages in the preclinical development of hemophilia gene therapy strategies has been the availability of several animal models for both hemophilia A and B. These models recapitulate many of the phenotypic aspects of human hemophilia and have proven to be very informative in exploring the efficacy and safety of gene therapy. Considerable progress has been made in the design of gene therapy protocols, and over the last 5 years it has been shown that long-term phenotypic correction, with sustained therapeutic levels of factor VIII (FVIII) and factor IX (FIX), can be attained in FVIII- and FIX-deficient mice and dogs using various viral vector-mediated gene therapy approaches. These animal models also have elucidated potential complications of gene therapy protocols, including acute vector-associated toxicities and the induction of neutralizing antibodies to the FVIII and FIX transgene products. Nevertheless, although the preclinical paradigm of hemophilic mouse followed by hemophilic dog studies has proven to be extremely helpful in evaluating the efficacy and safety of potential clinical gene therapy protocols, several limitations to these animal models still exist. This review presents a summary of the animal models available for hemophilia gene therapy, and highlights the various strengths and weaknesses of these models. PMID- 15118933 TI - Immune implications of gene therapy for hemophilia. AB - Similar to any novel treatment strategy for hemophilia, gene therapy faces the question of the risk of formation of inhibitory antibodies to the therapeutic factor VIII or factor IX protein. Activation of CD4 (+) or CD8 (+) T cells could lead to antibody formation or cytotoxic T lymphocyte responses to transgene expressing cells. Preclinical studies in animal models of hemophilia A and B with different mutations in the dysfunctional gene shed light on the risk for such immune responses and point toward strategies to avoid immune activation or even promote tolerance induction. The impacts of variables such as choice and design of gene transfer vector, underlying gene mutation, route of vector administration, and transient immune suppression are discussed. Maintenance of immunological hyporesponsiveness to the therapeutic gene product is critical for successful gene therapy. Recent studies provide evidence for tolerance induction to coagulation factor antigens by viral hepatic or neonatal in vivo gene transfer, by in utero gene delivery, and by oral or nasal administration of protein or peptides. PMID- 15118934 TI - Coagulation factors with improved properties for hemophilia gene therapy. AB - Hemophilias A and B are X-linked bleeding disorders that result in a qualitative or quantitative deficiency in coagulation factors VIII (FVIII) and IX (FIX), respectively. Affected patients experience significant morbidity as a result of repeated joint hemorrhages and subsequent arthropathy, and there is increased mortality related to life-threatening bleeding events. The mainstay of therapy is episodic or prophylactic infusions of plasma-derived or recombinant FVIII or FIX. However, gene transfer holds the promise of maintaining plasma levels of FVIII or FIX high enough to prevent the development of joint disease and reduce the risk of life-threatening bleeds or possibly even achieving normal plasma levels. Human gene therapy trials thus far have fallen short of this goal. This review summarizes the inherent limitations in expression of recombinant FVIII and the bioengineering strategies that are currently being explored for constructing novel recombinant FVIII molecules that have improved function. Current strategies for FVIII include increasing mRNA levels, improving secretion efficiency, increasing the rate of thrombin activation, stabilization of the activated form of FVIII, and strategies to prolong FVIII half-life in plasma by disrupting FVIII interaction with its clearance receptors. Strategies to improve the function of FIX include increasing the mRNA levels, reducing interaction with collagen IV, and increasing the specific activity. These novel molecules partnered with advances in gene transfer vector design and delivery may ultimately achieve persistent expression of FVIII and FIX, leading to an effective long-term treatment strategy for the hemophilias. PMID- 15118935 TI - Hemophilia gene transfer: comparison with conventional protein replacement therapy. AB - One of the unanswered questions in hemophilia is whether gene transfer, if successful, will be a safe and effective alternative to standard clotting factor treatment for hemophilia. Despite life-threatening complications of protein-based clotting factor treatment during the last three decades, including acquired immunodeficiency syndrome and hepatitis C, factor infusion is now considered safe, effective, and compatible with a normal lifespan. Thus, protein-based therapy will be the standard against which the safety and efficacy of gene transfer will be judged. Will the potential risks of gene transfer be sufficiently low to justify its use? Should all individuals with hemophilia consider gene transfer? To answer these questions, the known risks and benefits of current protein-based therapy must be compared with the potential risks and benefits of gene transfer. It is anticipated that risks of gene transfer may include the known risks of protein-based therapies, including allergic reactions, inflammatory responses, inhibitor formation, chronic hepatitis, as well as gene transfer-specific risks, including germline transmission, insertional mutagenesis, thrombosis, and potential ethical and psychological issues. This article reviews and compares the risks and benefits of standard protein-based therapy with those of gene transfer, and considers how gene transfer might fit into state-of-the-art management of hemophilia. PMID- 15118936 TI - Clinical gene transfer studies for hemophilia A. AB - The recent advances in gene transfer technology have expedited the development of gene therapy for the treatment of hemophilia A. Three different U.S. Food and Drug Administration-approved phase I clinical trials had been initiated using different gene therapy approaches each with their own advantages and limitations. In the first gene therapy trial for hemophilia A, a non-viral approach was being explored for patients with severe hemophilia A using ex vivo transfected dermal fibroblast expressing B-domain-deleted factor VIII ( BDD-FVIII). There were no serious adverse events and some patients appeared to have experienced fewer bleeding episodes with very low levels of FVIII near baseline. In the second trial, onco-retroviral vectors expressing BDD-FVIII were injected by peripheral intravenous infusion in adult patients suffering from severe hemophilia A. The procedure was safe and in some patients FVIII-transduced cells were detectable in the peripheral blood for more than a year. Although no sustained FVIII expression was detectable, occasional modest changes in FVIII levels were apparent, and in some cases a reduced bleeding frequency occurred compared with historical rates. In another trial, one patient suffering from severe hemophilia A has been treated with a high-capacity (or gutless) adenoviral vector expressing full-length FVIII, which appeared to have resulted in 1% of normal FVIII levels for several months. However, a transient inflammatory response with hematologic and liver abnormalities was observed. In conclusion, although modest improvements in clinical end points have been detected in some patients in these early phase I trials, further improvements in gene delivery technologies are warranted to bring hemophilia A gene therapy one step closer to reality. PMID- 15118937 TI - Clinical gene transfer studies for hemophilia B. AB - Hemophilia B, a deficiency of functional factor IX (FIX), has been extensively explored as a model for gene transfer. Two U.S. Food and Drug Administration approved clinical studies for hemophilia B have been undertaken, both using adeno associated viral vectors (AAV). AAV vectors have tropism for liver, muscle, central nervous system, and the respiratory tract; both skeletal muscle and liver have been used as target tissues in the hemophilia B studies. In both studies, proof of principle was first established in the hemophilia B dog model, with long term expression of canine FIX at therapeutic levels achieved before clinical studies were initiated. In the AAV-FIX muscle trial, vector was introduced into skeletal muscle of the upper and lower extremities of eight human patients by direct intramuscular injection. Muscle biopsies taken 2 to 10 months postinjection demonstrated gene transfer and expression (by Southern blot and immunofluorescence, respectively) in all patients, but circulating FIX levels were generally not >1%, and escalation of dose to levels that proved therapeutic in animals was thwarted by feasibility issues regarding the number of injections required. Nevertheless, the study demonstrated that parenteral injection of AAV FIX was safe at the doses tested, and could result in long-term expression of the transgene. Moreover, the general characteristics of transduction of human muscle were similar to those observed in other animal models. The safety and efficacy data established in the first trial formed the basis for a second trial in which AAV-FIX is administered systemically to target the liver. The liver study is currently ongoing, with six patients enrolled to date. PMID- 15118938 TI - The times they are a-changin': nontraditional treatment approaches to communication disorders. PMID- 15118940 TI - Nature-based therapy: its potential as a complementary approach to treating communication disorders. AB - Nature-based therapy (NBT) has been incorporated into the practice of many medical and mental health professions. Occupational therapists, physical therapists, nurses, social workers, psychologists, and to a lesser extent, speech language pathologists have used NBT practices as a complementary means of treating a variety of physical, emotional, and cognitive disorders. This article includes a description of NBT and the three types that comprise the practice and a review of the literature demonstrating the use of NBT in the general population and, more specifically, with individuals with neurogenic communication disorders. It concludes with a discussion of directions for future research of NBT. PMID- 15118941 TI - A review of select alternative treatment approaches for acquired neurogenic disorders: relaxation therapy and acupuncture. AB - Despite increasing consumer utilization of complementary and alternative medicine (CAM) and growing evidence of its positive effects on certain physical (e.g., chronic pain, insomnia) and psychiatric (e.g., anxiety) disorders, few empirical investigations have explored using CAM to treat neurogenic disorders of communication or cognition. Accordingly, to spur clinical and research interest in CAM, we review two forms of CAM, relaxation therapy and acupuncture, that have been used to address the needs of patients with neurogenic cognitive or communicative disorders. For each technique, we describe general therapy procedures, summarize and critique the extant research, and delineate areas in need of further empirical investigation. PMID- 15118942 TI - Exercise and volunteer work: contexts for AD language and memory interventions. AB - This article describes two novel contexts for language and memory stimulation in individuals with Alzheimer's disease (AD): student-supervised physical exercise and partnered volunteering at community agencies. Information presented is based on 5 years of the authors' experiences at the University of Arizona supervising student-administered interventions with mildly to moderately affected home dwelling AD patients in the Volunteers in Partnership and Elder Rehab programs. The authors report significant mood and fitness improvements by program participants, as measured on direct testing as well as by caregiver perceptions regarding the relative benefits of the four program components. Finally, they advocate for greater use of nontraditional approaches and utilizing students and volunteers when working with persons who have AD. PMID- 15118943 TI - Parkinson's disease: speech and voice disorders and their treatment with the Lee Silverman Voice Treatment. AB - Speech and voice disorders are very common among individuals suffering from idiopathic Parkinson's disease. In this article we review evidence for laryngeal, respiratory, articulatory, and velopharyngeal disorders in this population. We present the essential concepts and outcome data for the Lee Silverman Voice Treatment, a behavioral treatment program that has yielded significant long-term improvement in speech and voice functions in individuals with idiopathic Parkinson's disease. PMID- 15118944 TI - Transcranial magnetic stimulation as a complementary treatment for aphasia. AB - Functional brain imaging with nonfluent aphasia patients has shown increased cortical activation (perhaps "overactivation") in right (R) hemisphere language homologues. These areas of overactivation may represent a maladaptive strategy that interferes with, rather than promotes, aphasia recovery. Repetitive transcranial magnetic stimulation (rTMS) is a painless, noninvasive procedure that utilizes magnetic fields to create electric currents in discrete brain areas affecting about a 1-cm square area of cortex. Slow frequency, 1 Hz rTMS reduces cortical excitability. When rTMS is applied to an appropriate cortical region, it may suppress the possible overactivation and thus modulate a distributed neural network for language. We provide information on rTMS and report preliminary results following rTMS application to R Broca's area (posterior, R pars triangularis) in four stroke patients with nonfluent aphasia (5-11 years after left hemisphere stroke). Following 10 rTMS treatments, significant improvement in naming pictures was observed. This form of rTMS may provide a novel, complementary treatment for aphasia. PMID- 15118945 TI - Can drug therapies improve language functions of individuals with aphasia? A review of the evidence. AB - The neurochemistry of language and the neuropharmacology of aphasia are two domains of cognitive neuroscience still in their infancy. In this article we review what is known about these two domains, especially with regard to treating aphasia with drugs. Selected neurotransmitters can improve language function in certain patients with aphasia. We discuss which neurotransmitters work for which language functions in which patients, and why. PMID- 15118946 TI - [Action mechanisms of inhalation noxae in environment and occupation]. PMID- 15118947 TI - [Toxicological evaluation of inhalation noxae: test methods, assessment of toxic action and hazard potential, threshold limit values]. PMID- 15118948 TI - [Current therapy concepts in allergic and hyperergic diseases of the upper airways]. PMID- 15118949 TI - [Multiple chemical sensitivity (MCS) -- scientific and health policy aspects]. PMID- 15118950 TI - [Expert testimony problems in occupational diseases caused by inhalation noxae]. PMID- 15118951 TI - [Current viewpoints on hearing loss caused by occupational and leisure noise]. PMID- 15118952 TI - [Modern diagnostic methods for suspected glaucoma and glaucoma]. AB - BACKGROUND: Diagnosis of early glaucomatous damage as well as the detection of glaucomatous change are still difficult tasks. During the last years numerous new diagnostic techniques have been investigated and some of them have been introduced to the market. METHODS: This paper reviews the different aims of diagnostic technologies in the field of glaucoma. Methods appearing suitable for a large-scale use by ophthalmologists will be judged according to their suitability to meet the goals of early diagnosis, objective diagnosis, progression analysis and screening. CONCLUSIONS: The new diagnostic techniques have not yet greatly influenced our diagnostic procedure. This is mainly due to the fact that, for the most difficult borderline cases of glaucoma, these methods are of limited value, or have not yet been sufficiently investigated. Nevertheless, the Heidelberg retina tomograph (HRT), the nerve fiber analyzer GDX, short wavelength perimetry (SWAP), and the frequency doubling test (FDT) may play a role in these cases. For follow-up, HRT and GDx have proven valuable, especially in early stages of the disease. The very short testing time of FDT together with good specificity qualifies this test for glaucoma screening. PMID- 15118953 TI - [The headache patient at the ophthalmologist's]. AB - About 50 patients contact an ophthalmologist every month because of headache. Only 20 % can be helped by ophthalmological therapeutic efforts. The vast majority of those patients suffer from migraine. This contribution gives an overview of the most common causes of headache and their therapy. The ophthalmologist should not only exclude an ophthalmological cause of the headache but also contribute to making the correct diagnosis and initiating further treatment. PMID- 15118954 TI - [Acute therapy for eye burns]. AB - BACKGROUND: Eye rinsing is the major therapeutic measure with a decisive impact on prognosis after chemical or thermal eye irritation. Several different major targets are addressed by this measure. AIM OF THE STUDY: Up to now emphasis in research was placed mostly on the neutralisation of chemical products. Newer experimental results on the basis of biological experiments interpreted against the background of basic chemical reactions may enhance clinical treatment by introducing new mechanisms of intervention. MATERIALS AND METHODS: Experiments on the modification of osmolarity, buffering capacity and solubility products show new possibilities of therapy in eye rinsing after eye irritation. These mechanisms are demonstrated by means of key experiments. RESULTS: The official recommendation of rinsing a burnt eye with water seems to be insufficient against the background of actual experimental data. Measurements of pH after eye burns indicate no buffering effects for water, or aqueous saline solution. There was weak buffering for phosphate buffer in alkali burns and a high capacity for neutralisation for diphoterine in alkali and acid burns. Ionic contents and osmolarities of the rinsing solutions have a decisive influence on the ionic composition and osmolarity of the burnt cornea after rinsing. Cellular damage is enhanced in unaffected healthy cell cultures by hyposomolar rinsing. CONCLUSION: We recommend buffered solutions with high buffer capacities for initial eye rinsing. The advantage or disadvantage of the elevated concentration of ions in the buffered rinsing solutions determining osmolarity cannot be confirmed or refuted up to now. It seems to us to be certain that osmolarity is a decisive future factor in initial rinsing. PMID- 15118955 TI - [Rotational stability of the eye in standard photography]. AB - BACKGROUND: After the implantation of a toric IOL (tIOL), postoperative rotations of more than 30 degrees have been reported. Beyond this scope, we investigated the reproducibility of the eye's orientation (rotation stability) in principle. PATIENTS AND METHODS: The rotational stability of the eye was investigated using standard fundus photographs (telecentric fundus camera, Zeiss, Oberkochen). One hundred eyes of 50 patients (28 females) were photographed in a time interval of more than 6 months twice. With the aid of significant markers on the fundus photograph, the axial position of the eye was defined and the rotational angle between the two slides of one eye was measured. RESULTS: The mean absolute rotational angle was 2.4 +/- 1.7 degrees (range 0 to 7.5 degrees ) in all 100 eyes (2.5 +/- 1.6 degrees right eye, 2.4 +/- 1.7 degrees left eye). Only 6 % of the eyes did not rotate. A range of less than 3 degrees rotation was detected in 52 % of the eyes. A rotation of equal to or more than 3 degrees was noted in 42 %. The patient's age did not influence the amount of globe rotation. In- and excyclorotations showed a coincidental distribution and no side differences. CONCLUSIONS: Seven and a half degrees or 3 degrees tIOL rotation would correspond to a residual astigmatism of 26 % or 10 %. When measuring the marked axis of the tIOL, these results are markedly influenced by head inclination, rotation of the head, incyclorotation or excyclorotation. This is transferable, for example, to autorefractor, corneal topography and sectorial analyses for glaucoma diagnostics. PMID- 15118956 TI - [Intraocular non-Hodgkin's lymphoma and its therapy-- a case series of ten patients]. AB - BACKGROUND: The timely and correct diagnosis of intraocular non-Hodgkin's lymphoma represents a huge challenge to clinicians. Two thirds of intraocular lymphomas are a manifestation of a primary CNS lymphoma (PCNSL) arising outside the lymphatic system and are localized in the brain, the meninges or the spinal chord. Ten to twenty percent commence as vitreous or retinal infiltrates mimicking uveitis. Ninety five percent of PCNSL are B cell lymphomas. PATIENTS: Three exemplary cases from a group of ten patients treated between 1998 and 2002 are presented. A table provides a summary of the relevant details of all ten patients. RESULTS: The mean age at presentation was 63.5 years with a female to male ratio of 6 to 4. Nine patients were diagnosed as having intermediate or posterior uveitis, in one patient choroidal metastases were suspected. Six patients had a concomitant CNS lesion while four patients showed isolated intraocular lymphoma only. The presence of a highly malignant B cell lymphoma was proven by vitreous biopsy in nine cases and by stereotactic biopsy of a CNS lesion in one patient. All patients were treated by intravenous chemotherapy, however, no binding recommendations with regard to treatment exist to date. CONCLUSIONS: We give an overview of all current treatment regimens and their pitfalls. At present it is recommended that all patients with proven PCNSL be entered in a multicenter randomized study under the auspices of the Department of Internal Medicine III of the Benjamin-Franklin-University-Hospital, Berlin and the Department of Neurology of the University Hospital of Tuebingen. PMID- 15118957 TI - [Spontaneous dislocation of intraocular lens with capsule as a late complication of cataract surgery in patients with pseudoexfoliation syndrome --five case reports]. AB - PURPOSE: Pseudoexfoliation syndrome (PEX) is associated with zonular weakness and a higher frequency of intraoperative complications during cataract surgery, including rupture of the posterior lens capsule, zonular dialysis and a rise of intraocular pressure occurring postoperatively. Delayed dislocation of an IOL is a rarely reported phenomenon. PATIENTS: Within one year, late dislocation of the lens capsule with the in the bag fixated IOL was observed following cataract surgery in five patients (67, 74, 79, 90 and 92 years old) with pseudoexfoliation syndrome. RESULTS: All patients had an uneventful in the bag implantation of the IOL 6 (three patients), 3 and 11 years ago, respectively. Postoperatively occurring secondary cataract was treated by a YAG-capsulotomy in four cases. No patient had any other predisposing factors that would lead to zonular weakness besides the pseudoexfoliation syndrome. The dislocation of the IOL and capsule occurred spontaneously. In one patient with preexisting glaucoma, the dislocation was followed by an increase of intraocular pressure. All cases were successfully treated with IOL explantation, anterior vitrectomy and placement of an anterior chamber IOL. CONCLUSION: Patients with pseudoexfoliation syndrome undergoing cataract surgery may be at risk not only for intraoperative complications but also for delayed spontaneous dislocation of the IOL and capsule. This possible complication should be considered in surgical planning for patients with pseudoexfoliation syndrome. In these patients it may be better to implant the IOL in the ciliary sulcus. PMID- 15118958 TI - [Phacogenic uveitis with luxation of of Soemmering's ring formation in the vitreous body 63 years after cataract surgery]. AB - BACKGROUND: After extracapsular cataract extraction, the remaining epithelial cells near to the equator may lead to the formation of a Soemmerings ring. Only 4 cases of dislocation of the ring in the vitreous are reported in the literature. A spontaneous dislocation associated with a phacolytic uveitis has not been described before. HISTORY AND SIGNS: In 1999 a 68-year-old man attended our clinic with a vision loss from 0.3 to hand movements in his left eye. For some years his right eye had been blind after unsuccessful retinal detachment surgery. In his left eye conjunctival inflammation, anterior chamber flare and complete occlusion of the pupil were found. Intraocular pressure was normal under hypotensive medication. Ophthalmoscopy was not possible and ultrasound revealed a ring-like dense structure in the vitreous cavity. THERAPY AND OUTCOME: Pars plana vitrectomy showed a Soemmerings ring, which was successfully removed by pars plana fragmentation. Forty-eight months postoperatively the patient had a visual acuity of 0.2 in a healthy and normotensive eye. CONCLUSIONS: Even years after cataract extraction a spontaneous dislocation of Soemmerings ring is possible. PMID- 15118959 TI - [Bilateral visual field defects with optic disc drusen and secondary open angle glaucoma with PEX--clinical correlation with the HRA]. AB - BACKGROUND: Arcuate visual field defects are a typical sign of glaucomatous damage. Elevated intraocular pressure in combination with pseudoexfoliation syndrome (PSX) manifests the diagnosis glaucoma. Beyond this state, in microdiscs with optic disc drusen, the exact classification of the visual field defects is crucial. CASE REPORT: A 57-year-old male with pseudoexfoliation glaucoma was referred because of progressive glaucomatous visual field defects. The visual acuity was right 20/40 and left 20/25. Maximum intraocular pressure was 36 mm Hg. A simple optic nerve atrophy was diagnosed superonasally. The optic disc size was OD 2.24 mm(2) and OS 1.89 mm(2) (HRT I). An Ultrasound B-mode scan demonstrated the diagnosis of optic disc drusen. Over a follow-up of 1 year, a growth tendency was observed, especially in the superonasal quadrant. The mulberry-shaped surface of the drusen was visualized with infrared reflection images (HRA II, 830 nm). Confocal scanning laser ophthalmoscopy (HRA II, excitation 488 nm, 500 nm notch filter) showed an increased intrapapapillary autofluorescence (> 50 % papillary area: OD 1.67 mm(2), OS 1.26 mm(2)). This technique could detect drusen in areas that looked normal in classical retinoscopy. CONCLUSION: The differential diagnosis of arcuate scotomas includes simple optic nerve atrophy and glaucomatous optic nerve atrophy. Optic disc drusen in glaucoma eyes can obscure the main cause of progressive visual field loss. Superficial optic disc drusen can be measured planimetrically over the years. An adequate reduction of intraocular pressure should be realized in these eyes. PMID- 15118961 TI - Cerebral vasoconstriction syndromes. AB - Stroke from reversible cerebral arterial vasoconstriction has been described in a variety of conditions, including migraine, pregnancy, puerperium (postpartum angiopathy), use of vasoconstrictive drugs, Call-Fleming syndrome, and benign angiopathy of the central nervous system. Although vasoconstriction is an important cause of ischemic and hemorrhagic stroke in young individuals, vasoconstriction syndromes have not been well characterized and remain under recognized. Misdiagnosis is common because the clinical and radiological features can overlap with conditions such as primary cerebral vasculitis. With the advent of newer, noninvasive angiography techniques and the escalating use of vasoactive drugs, it is likely that clinicians will encounter more patients with vasoconstriction-induced stroke. This article reviews the history, clinical and radiological characteristics, differential diagnosis, and management of cerebral vasoconstriction syndromes. PMID- 15118962 TI - Hemicraniectomy for massive cerebral infarction. AB - The most important acute phase complication of massive cerebral infarction is cerebral herniation secondary to brain edema. In the past decade, there has been heightened interest in surgical treatment of this problem with the use of the techniques of hemicraniectomy and dural augmentation. In this article, we review the theoretical and clinical data supporting the use of hemicraniectomy in malignant cerebral infarction and the range of outcomes expected in patients who undergo this procedure. PMID- 15118963 TI - Motor recovery strategies after stroke. AB - Impaired motor function after stroke is a major cause of disability in young stroke survivors. The plasticity of the adult human brain provides opportunities to enhance traditional rehabilitation programs for these individuals. Younger stroke patients appear to have a greater ability to recover from stroke and are likely to benefit substantially from treatments that facilitate plasticity mediated recovery. The use of new exercise treatments, such as constraint-induced movement therapy, robot-aided rehabilitation, and partial body weight supported treadmill training are being studied intensively and are likely to ultimately be incorporated into standard poststroke rehabilitation. Medications to enhance recovery, growth factors, and stem cells will also be components of rehabilitation for the young stroke survivor in the foreseeable future. PMID- 15118964 TI - Age and functional outcome after stroke. AB - The effect of age on functional outcome after stroke remains uncertain. Many studies have found that younger patients do better than older patients, whereas others have found minimal or no effect of age on rehabilitation outcomes. We examined the effect of advancing age on FIM trade mark gain, length of stay, length of stay efficiency, and home discharge in 979 stroke rehabilitation patients at a long-term acute care rehabilitation hospital. We found a strong relationship of increasing age to poorer outcome in all measures for patients with admission FIM (AFIM) score <40, a variable relationship in those with AFIM 40-80, and no relationship of age to the outcome measures in patients with AFIM >80. PMID- 15118965 TI - Computer adaptive testing: a strategy for monitoring stroke rehabilitation across settings. AB - Current functional assessment instruments in stroke rehabilitation are often setting-specific and lack precision, breadth, and/or feasibility. Computer adaptive testing (CAT) offers a promising potential solution by providing a quick, yet precise, measure of function that can be used across a broad range of patient abilities and in multiple settings. CAT technology yields a precise score by selecting very few relevant items from a large and diverse item pool based on each individual's responses. We demonstrate the potential usefulness of a CAT assessment model with a cross-sectional sample of persons with stroke from multiple rehabilitation settings. PMID- 15118966 TI - Preventing a second stroke in the young. AB - In the United States, more than 220,000 adults under the age of 45 have had a stroke. Secondary stroke prevention is critical in this population because of the potential impact of a second stroke on productivity and future quality of life. In addition to conventional stroke risk factors, younger patients have special risk factors inherent to genetic and environmental elements, such as the use of illicit drugs and cardiac abnormalities; in women, risk factors include pregnancy, migraine, and the use of oral contraceptives. PMID- 15118967 TI - Long-term functional outcome of pediatric stroke survivors. AB - PURPOSE: To examine the long-term functional, psychosocial, and medical outcome of pediatric stroke survivors. METHOD: This was a descriptive survey performed on patients with childhood stroke who participated in an earlier study. Measures included the Vineland Adaptive Behavior Scales (VABS) and the Diener Satisfaction with Life Scale. Current information on living situation, school placement, employment, and medical outcome were obtained. RESULTS: Twenty-nine (58%) patients participated. The mean age was 19.3 years (SD = 6.6), mean age of onset of stroke was 7.0 years (SD = 5.4), and mean follow-up time was 11.9 years (SD = 3.9). Diagnoses included hemorrhagic (31%) and ischemic (69%) stroke. All but one adult had finished high school, and the majority of participants had gone to college. 60% of patients over age 16 were employed. The average VABS levels for communication, daily living skills, socialization, and adaptive behavior fell into the moderately low range. Use of seizure medications and ADL dependence were the predictors for lower VABS levels (p <.05). Younger age, ischemic stroke, and previous dependence in mobility were risk factors for lower scores for the self care domain, but not for lower life satisfaction. Patients who scored below adequate on VABS tended toward lower life satisfaction. CONCLUSION: Pediatric stroke survivors had good educational and mobility outcomes, but communication, ADL, and socialization fell into the low-moderate range. The different predictors of functional and subjective quality of life outcomes suggest that functional outcomes may mediate the relations between medical factors and satisfaction with life. PMID- 15118968 TI - Closing the gap between research and clinical practice. AB - Evidence-based practice and the application of research findings to practice is a major focus of attention in health care today. The quantification of variations in service delivery and the economic implications of these variations contribute to the pressure on clinicians to provide evidence-based care whenever possible. Awareness of the development of evidence-based practice (EBP), the need for EBP, factors that affect EBP, the methods used to translate research into practice, EBP quality indicators, barriers to EBP, and benefits of EBP will assist clinicians in the effective use of evidence-based information in the care of their patients. PMID- 15118969 TI - Concepts of medical necessity in rehabilitation. PMID- 15118970 TI - What are male caregivers talking about? AB - PURPOSE: Using secondary analysis and Friedemann's framework of systemic organization as a guide, the purpose of this article is to examine the experience of a subsample of five men caring for women with stroke who participated for 3 months in a Web-based support group. METHOD: Analysis of qualitative data (N = 224 entries) obtained from interview questions and discussion amongst the caregivers followed established protocols. RESULTS: Results revealed that these men were dealing with role changes and women's depression and irritability. The stroke event also fostered closer family ties and "being there" for one another. CONCLUSION: These findings begin to explore the complex experience of men caring for women with stroke. PMID- 15118971 TI - Considering powered mobility for individuals with stroke. AB - Power mobility is an option that provides many disabled individuals with a means of locomotion. Stroke is the leading cause of long-term disability in this country. Unfortunately, there is currently a lack of published material that expounds on power mobility as it relates to clients who have had a stroke. We will present the benefits and considerations for choosing power wheelchairs for this client population. A brief review of seating and mobility evaluation and funding considerations is included with an emphasis on specific issues concerning stroke clients. Finally, a call for further research on this topic is presented. PMID- 15118972 TI - Functional abdominal pain syndrome. AB - Functional abdominal pain syndrome (FAPS) is one of the less common functional gastrointestinal disorders, yet it is one with high health care impact. It is best understood from a biopsychosocial perspective as the interaction of biologic and psychosocial factors contributing to symptom experience and behavior. The pain of FAPS relates primarily to dysfunction of central pain modulatory systems; however, disturbances in motility or visceral hypersensitivity may contribute. There are characteristic clinical and behavioral features that typify patients with FAPS, and the medical evaluation must include their recognition through psychosocial assessment, physical examination, and a conservative, cost-effective effort to exclude other diseases. The treatment approach depends on establishing an effective physician-patient relationship, setting reasonable treatment goals, and using multicomponent medical and behavioral strategies that are individualized to patient needs. In refractory cases, referral to a multidisciplinary pain treatment center might be helpful. PMID- 15118973 TI - Recognition of genetic syndromes in families with suspected hereditary colon cancer syndromes. PMID- 15118974 TI - 5-ASA therapy for active Crohn's disease: old friends, old data, and a new conclusion. PMID- 15118975 TI - Oral Pentasa in the treatment of active Crohn's disease: A meta-analysis of double-blind, placebo-controlled trials. AB - BACKGROUND & AIMS: The aim of this study was to perform a meta-analysis of the efficacy results from 3 placebo-controlled multicenter clinical trials of slow release mesalamine (Pentasa) for the acute treatment of mild to moderate Crohn's disease. METHODS: Three trials fulfilled the selection criteria (double-blind, placebo-controlled, randomized studies in adult patients treated with Pentasa 4 g/day for active Crohn's disease). The efficacy and safety was evaluated in these trials by using the Crohn's Disease Activity Index (CDAI) as the primary efficacy variable. The study duration was 16 weeks in all 3 trials. The total numbers of patients were 304 in the Pentasa 4-g/day treatment groups and 311 in the placebo groups. A meta-analysis was performed based on the study reports. RESULTS: For the intent-to-treat patients in the Pentasa groups, the overall mean reduction of the CDAI from baseline to the final visit was -63 points. The corresponding CDAI change in the placebo groups was -45 points; the net difference was -18 points. Compared with placebo, the 4-g/day dose of Pentasa was associated with a statistically significant overall improvement in the CDAI from baseline to the final visit (P = 0.04). When the meta-analysis was restricted to protocol correct patients, the effect of Pentasa became more pronounced (overall mean reduction of -83 CDAI points; P = 0.02, compared with placebo). Contrary to the consistent effects with Pentasa, the trial-specific reductions of the CDAI with placebo differed significantly between the trials. CONCLUSIONS: The meta-analysis of 3 large, double-blind, randomized studies in the treatment of active Crohn's disease confirms that Pentasa 4 g/day is superior to placebo in reducing the CDAI but the clinical significance of the magnitude of this difference is not clear. PMID- 15118976 TI - Predictors of outcome of pneumatic dilation in achalasia. AB - BACKGROUND & AIMS: Graded pneumatic dilation (PD) is a widely accepted treatment for achalasia. We investigated the potential predictors of outcome in a large group of patients with achalasia and tested the hypothesis that graded PD may not be appropriate for all patients. METHODS: Patients undergoing PD from 1992 to 2002 were evaluated retrospectively. Symptom scores (0-15) for dysphagia (0-5), regurgitation (0-5), and chest pain (0-5), as well as degree of esophageal emptying by timed barium swallow, were assessed for all patients. Failure was defined as the return of symptoms resulting in repeated PD or surgical myotomy. Clinical data assessed for short- and long-term predictors of response. RESULTS: Seventy-five patients with achalasia without previous therapy constituted the studied population. Three-year success rates for PD using 3.0-cm, 3.0-cm followed by 3.5-cm, and 3.0-cm and 3.5-cm followed by 4.0-cm Rigiflex balloons were 37% (95% confidence interval [CI], 26-53), 76% (95% CI, 65-88), and 88% (95% CI, 80 97), respectively. Patient age and sex were important treatment outcome predictors. A Cox proportional hazards model of time to additional therapy on sex and 10-year increase in age showed that 3.0-cm PD was significantly (P = 0.04) more likely to fail in younger men than older men (hazard ratio, 0.63; 95% CI, 0.41-0.98). In 25 of 68 patients (37%) initially treated with a 3.0-cm balloon, PD failed within 3 months. Twenty-two of 25 patients (88%) with early failure were men. CONCLUSIONS: (1) Young men have a greater failure rate with 3.0-cm PD than older men or women in general, and (2) graded PD in this group starting initially with the 3.0-cm balloon is more likely to fail. PMID- 15118977 TI - Chronic abdominal wall pain: clinical features, health care costs, and long-term outcome. AB - BACKGROUND & AIMS: Chronic abdominal wall pain (CAWP) often is misdiagnosed. We evaluated CAWP patients regarding diagnosis accuracy, clinical features, comorbidity, referral frequency, use of care, and long-term outcome. METHODS: We reviewed the records of all outpatients referred to a gastroenterologist in 5 years, recorded referral indications, and identified patients initially diagnosed with CAWP or irritable bowel syndrome (IBS). Charts of all CAWP patients were reviewed, and direct costs were estimated for abdominal pain-related physician visits and imaging studies during the 12 months before and 12 months after consultation. We appraised long-term pain status by telephone. RESULTS: Of 2709 patients, CAWP was diagnosed by physical examination in 137 patients; the diagnosis remained unchanged after 47.3 +/- 17.7 (mean +/- SD) months in 133 (97.1%) patients. Women predominated over men 4 to 1, pain was usually upper abdominal, had lasted 25.3 +/- 46.3 months, and obesity and painful comorbidities and depression were common. CAWP and IBS comprised 7.8% and 16.3% of symptomatic referrals, respectively. Prereferral, physicians rarely suspected CAWP and often prescribed therapy for acid-peptic disease. Postconsultation, primary care, emergency and specialist visits, and radiologic examinations markedly decreased (P < 0.001), and estimated annual costs decreased from $1133.87 +/- 953.37 to $541.33 +/- 989.04 (P < 0.0001). Therapy varied, and 44 (47.3%) patients had no pain at follow-up evaluation. CONCLUSIONS: CAWP is a common underrecognized disorder. Comorbidities are frequent, and health care use is high. Diagnosis is accurate and reduces health care costs. Over the long term, pain disappearance and persistence occurs in approximately equal proportions of patients. PMID- 15118978 TI - Improvement of collateral vessels in the vicinity of gastric cardia after endoscopic variceal ligation therapy for esophageal varices. AB - BACKGROUND & AIMS: Endoscopic variceal ligation (EVL) therapy has been performed widely to treat or prevent variceal bleeding. We sought to examine the influence of EVL for esophageal varices on collateral vessels in the vicinity of gastric cardia. METHODS: In 42 patients with esophagogastric varices, conventional endoscopy and endoscopic ultrasonography with a 20-MHz probe (CUP-EUS) were performed before and at every 3 months after EVL for esophageal varices. By using conventional endoscopy, cardial variceal sizes were divided into 3 grades: F0, F1, and F2. The sizes of submucosal, perforating, and paracardial vessels at the cardia also were classified into 3 grades according to CUP-EUS findings. RESULTS: Conventional endoscopy showed cardial varices in 33 (79%) patients before and 23 (55%) patients at 3 months after the treatment (P < 0.05). CUP EUS showed that 29 (69%) patients had severe grade cardial submucosal vessels before EVL, but only 13 (31%) patients did after the treatment (P < 0.01). Nineteen (45%) patients had severe grade cardial perforating vessels before EVL, but only 4 (10%) patients did after the treatment (P < 0.001). Furthermore, patients with severe grade residual submucosal or perforating vessels at the cardia had shorter recurrence free times of esophageal varices (P < 0.01, 0.05, respectively). CONCLUSIONS: Collateral vessels in the vicinity of gastric cardia were improved significantly after EVL, indicating that esophageal varices can be treated by EVL even though they connect with cardial varices. Furthermore, eradication of such collateral vessels by EVL may lead to longer recurrence-free status of esophageal varices. PMID- 15118979 TI - A prospective, quantitative assessment of the effect of ethanol and other variables on the endosonographic appearance of the pancreas. AB - BACKGROUND & AIMS: Studies comparing endosonography with endoscopic pancreatography, histological examination, and functional assays show that the number of endosonographic abnormalities (or "criteria") increases with the severity of chronic pancreatitis. However, it is unclear to what extent such variables as demographics, body habitus, and routine exposure to such pancreatic toxins as ethanol and cigarette smoke can affect pancreatic endosonography. The aim of the study is to quantify the effects of these variables precisely. METHODS: Pancreatic endosonography was performed by a single operator in consecutive patients referred for any indication. The relationship between 8 possible endosonographic criteria and these variables was studied. RESULTS: One thousand one hundred fifty-seven patients were studied (93% of eligible patients). Number of criteria correlated most strongly with ethanol ingestion (r = 0.273; P = 0.0001) and smoking history (r = 0.201; P = 0.0001). It did not correlate with age or body mass index. The strongest independent predictors of severe pancreatic abnormalities (>/=5 criteria) were heavy ethanol ingestion (odds ratio [OR], 5.1; 95% confidence interval [CI], 3.1-8.5), male sex (OR, 1.8; 95% CI, 1.3-2.55), clinical suspicion of pancreatic disease (OR, 1.7; 95% CI, 1.2 2.3), and heavy smoking (OR, 1.7; 95% CI, 1.2-2.4). Severe endosonographic abnormalities were found in only 2 of 51 patients (3.9%) with no risk factors or symptoms of pancreatic disease. CONCLUSIONS: Several variables can affect the endosonographic appearance of the pancreas independently. Severe abnormalities may be asymptomatic. The clinical, functional, and histological significance of endosonographic abnormalities requires clarification. PMID- 15118980 TI - Thiopurine methyltransferase activity influences clinical response to azathioprine in inflammatory bowel disease. AB - BACKGROUND & AIMS: Genetic polymorphism in thiopurine methyltransferase (TPMT) activity may influence clinical responsiveness to azathioprine (AZA) therapy. Our aim was to determine if the measurement of erythrocyte TPMT enzyme activity could be used to optimize clinical responsiveness to AZA therapy in patients with inflammatory bowel disease (IBD). METHODS: A total of 142 consecutive patients were studied. Forty-one patients (32 with Crohn's disease [CD] and 9 with ulcerative colitis [UC]) were enrolled in a 4-month prospective nonrandomized study with AZA, and 101 (65 with CD and 36 with UC) were on either maintenance AZA or 6-mercaptopurine (6-MP). Erythrocyte TPMT activity and AZA metabolite levels were measured blinded to the clinical response. RESULTS: The response rate after 4 months of continuous AZA therapy was 69% (9/13) in those patients with below-average (12 U/mL blood (P < 0.001). Patients with TPMT activity 12 (218 +/- 28), despite similar mean (1.6 mg/kg/day) dosages of AZA (P < 0.001). By multivariate logistic regression analysis, patients with a TPMT level <15.3 U/mL blood were 6.2 times more likely to respond to AZA therapy. A 6-TGn level of >292 pmol/8 x 10(8) RBCs was associated with a positive predictive value of clinical response of 85.7%. CONCLUSIONS: Patients with higher than average TPMT activity (>12) may remain refractory to conventional dosages of AZA, and may require high (>292) 6-TGn levels. Prospective, randomized, controlled trials are needed to determine whether prior TPMT phenotype testing can be used to adjust the dose of AZA effectively to improve clinical response time and rate. PMID- 15118981 TI - Galactose elimination capacity as a prognostic marker in patients with severe acetaminophen-induced hepatotoxicity: 10 years' experience. AB - BACKGROUND & AIMS: Patients with acetaminophen-induced fulminant hepatic failure may have the capacity for recovery if sufficient liver cell mass remains to allow regeneration. We investigated the prognostic potential of the galactose elimination capacity (GEC) as a noninvasive measurement of functioning liver cell mass in severe acetaminophen-induced hepatotoxicity. METHODS: All patients admitted with acetaminophen poisoning during a 10-year period were studied retrospectively. A total of 220 patients who had at least one GEC performed were included in the study. RESULTS: The GEC was lower in patients with than without hepatic encephalopathy (14.5 +/- 5.6 micromol/min/kg vs. 23.2 +/- 6.7 micromol/min/kg; P < 0.0001). Among patients with hepatic encephalopathy, the GEC was significantly higher in spontaneous survivors than in nonsurvivors (16.8 +/- 5.6 micromol/min/kg vs. 12.2 +/- 4.7 micromol/min/kg; P < 0.0001). In a logistic regression analysis, GEC was associated independently with mortality (odds ratio: 1.28 per 1 micromol/min/kg decrease in GEC; 95% confidence interval: 1.14-1.45). A threshold GEC of 16.5 micromol/min/kg to identify nonsurvivors had a sensitivity of 90%, a specificity of 72%, a positive predictive value of 49%, and a negative predictive value of 96%. None of 14 patients with hepatic encephalopathy and a GEC less than 10 micromol/min/kg survived. CONCLUSIONS: The GEC was strongly associated with development of hepatic encephalopathy and death from acetaminophen-induced fulminant hepatic failure. The GEC was too unspecific to be used alone for identification of transplantation candidates, but it may be useful as a supplement to other selection criteria. PMID- 15118982 TI - Effectiveness of interferon alpha-2b and ribavirin combination therapy in the treatment of naive chronic hepatitis C patients in clinical practice. AB - BACKGROUND & AIMS: Studies in many diseases have shown that efficacy in clinical trials often does not translate into effectiveness in clinical practice. The aims of this study were to determine the rate of sustained virological response (SVR) and the factors associated with SVR in therapy naive chronic hepatitis C patients treated with interferon alpha-2b and ribavirin combination therapy at a university outpatient clinic. METHODS: The medical records of 153 consecutive chronic hepatitis C patients treated between June 1998 and May 2001 were reviewed. RESULTS: The mean subject age was 44 years, 64% were men, 85% were white, 56% had HCV genotype 1, and 21% had cirrhosis on biopsy. The overall SVR rate was 42% (29% in genotype 1/4; 65% in genotype 2/3). Side effects resulted in interferon or ribavirin dose reductions in 22% of patients and premature termination of treatment in 10%. The SVR rate was significantly higher in the 102 patients who received >80% of the recommended dose and duration of therapy compared with the 51 patients who did not (53% vs. 20%, P = 0.00008). HCV genotype, subject race, and adherence were independently associated with SVR (P < 0.01). Although the incidence of side effects and medication adherence was similar in blacks and whites, adherent blacks had a significantly lower SVR rate (14% vs. 58%, P < 0.01). CONCLUSIONS: Despite the inclusion of a broader spectrum of patients and less frequent monitoring, combination antiviral therapy in our treatment-naive chronic hepatitis C patients was of similar efficacy to that reported in large multicenter trials. In addition, our data show that medication adherence is an important predictor of SVR in an academic clinical practice. PMID- 15118983 TI - Impact of highly active antiretroviral therapy on the spectrum of liver disease in HCV-HIV coinfection. AB - BACKGROUND & AIMS: Hepatitis C virus (HCV) coinfection is common in patients with human immunodeficiency virus (HIV) infection. The mortality associated with HIV has decreased dramatically with the introduction of highly active antiretroviral therapy (HAART). However, the impact of HAART, including protease inhibitors (PIs), nucleoside reverse-transcriptase inhibitors (NRTIs), and non-NRTIs (NNRTIs), on the spectrum of HCV-related liver disease remains unclear. The purpose of this retrospective analysis is to determine the impact of PI and NNRTI use on liver histological characteristics in patients with stable HIV-HCV coinfection (n = 101) compared with HIV-uninfected controls with HCV infection (n = 302). METHODS: The majority of coinfected patients were men (75%), African American (82%), and had genotype 1 HCV (91%). Mean HIV load was 1.52 log copies/mL, 48% had undetectable HIV RNA and a mean CD4 count of 528 cells/microL, and 11% had a CD4 count < 200 cells/microL. Both mean alanine aminotransferase (ALT) level (83 U/L; 54% had a normal ALT level) and Knodell Histological Activity Index score (7.04; 33% had advanced fibrosis) were similar to those of our control population. Ninety-three percent of patients were administered a mean of 3 antiretroviral medications: NRTIs in 98%, NNRTIs in 45%, and PIs in 54%. RESULTS: There were no significant differences in biochemical or histological parameters between patients administered or not administered PIs or NNRTIs. CONCLUSIONS: In this uncontrolled retrospective analysis, we were unable to show a significant impact of either PI or NNRTI use on the spectrum of liver disease. PMID- 15118984 TI - Differential efficacy of corticosteroids and interferon in a patient with chronic hepatitis C-autoimmune hepatitis overlap syndrome. AB - The chronic hepatitis C-autoimmune hepatitis (AIH) overlap syndrome has been described in the literature, but to date appropriate therapy remains controversial. We report on a 28-year-old woman with hepatitis C-AIH overlap syndrome. The patient was infected with HCV genotype 1b and had laboratory and immunologic findings of AIH type 2 such as increased Igs and a high titer of antibodies against liver-kidney microsomes. Initial liver biopsy specimen demonstrated end-stage liver fibrosis due to chronic hepatitis. After long lasting corticosteroid treatment, only partial remission was achieved. In contrast, short-term antiviral therapy with interferon-alpha2b in combination with ribavirin was followed by complete biochemical and virologic remission. However, 15 months later, a relapse of AIH was observed. After restarting corticosteroid treatment, transaminase levels completely normalized. Surprisingly, in this patient with overlap syndrome, short-term interferon therapy induced complete remission of chronic HCV infection and regression of severe liver fibrosis. PMID- 15118985 TI - Hemorrhagic shock due to mesenteric inflammatory veno-occlusive disease: An underdiagnosed cause of intestinal ischemia? PMID- 15118986 TI - The relationship of anger expression and alexithymia with coronary artery stenosis in patients with coronary artery diseases. AB - This study examined the relationship between anger expression or alexithymia and coronary artery stenosis in patients with coronary artery diseases. 143 patients with coronary artery diseases (104 males and 39 females) were enrolled in this study. The severity of their coronary artery stenosis was measured by angiography. The Anger Expression Scale and the Toronto Alexithymia Scale were used to assess the level of anger expression and alexithymia. The more stenotic group (occluded by 75% or more) exhibited a significantly higher level of alexithymia than the less stenotic group (occluded by less than 25%). Multiple regression analysis on the extent of stenosis also revealed that regardless of gender and age, the coronary artery disease patients with higher alexithymia were likely to show a greater level of stenosis. However, no significant differences were found on either the anger-in or anger-out subscale scores between the two groups. These results suggest that alexithymia is associated with the severity of coronary artery stenosis in patients with coronary artery disease. However, both anger expression and anger suppression were not shown to be associated with the severity of coronary artery stenosis. PMID- 15118987 TI - Apoptosis and proliferation in paired primary colorectal adenocarcinomas and their liver metastases. AB - The proliferation potentials and the level of apoptosis were compared in paired primary colorectal adenocarcinomas and their liver metastases within each individual. From a total of 22 patients 44 specimens of paired primary and metastatic tumors were obtained for analysis. The levels of spontaneous apoptosis (a spontaneous apoptosis index, SAI: % apoptotic nuclei among a total of 1000 nuclei) and of proliferation (KI-67 index: % positively stained cells for KI-67 among a total of 1000 cells) were analyzed between primary and metastatic tumors. Survival rates and its relationship with the clinical parameters were also analyzed. The overall survival rate at 5 years was 16.9% with the median survival time of 45 months. T-stage (p=0.005) and time to liver metastasis (synchronous versus metachronous, p=0.03) showed statistical significance in relation to survival. The mean SAI of primary tumors was 1.35 +/- 0.25, which was not statistically different from the 1.58 +/- 0.18 of metastatic tumors (p=0.33). The mean KI-67 indices in primary and metastatic tumors were 23.9 +/- 3.4 and 16.4 +/ 2.5, respectively, and this difference was statistically significant (p=0.016). Subset analysis showed significant difference in the KI-67 index in the synchronous group but not in the metachronous group. No significant difference was shown in the relative ratios of apoptosis to proliferation between the primary tumor and the metastasis within each individual. The results in this study may partly explain the indolent behavior of liver metastasis from colorectal cancer and provides a rationale for the active treatment of metastatic tumors as well as of primary disease. PMID- 15118988 TI - Discharge decision-making by intensivists on readmission to the intensive care unit. AB - Patients readmitted to the intensive care unit (ICU) have a significantly higher mortality rate. The role of intensivists in judging when to discharge patients from the ICU is very important. We undertook this study to evaluate the effect of the intensivists' discharge decision-making on readmission to ICU. The intensivists actively participated in the discharge decision-making, with the discharge guideline taken into consideration, in respect of group 1 patients, but not in respect of group 2. The readmission rate in group 1 was lower than that in group 2. The readmission in patients in each group was associated with higher mortality rates and longer lengths of stay at the ICU. Respiratory failure was the major cause of readmission. In the non-survivors out of the readmitted patients, the Acute Physiology and Chronic Health Evaluation (APACHE) scores on the initial discharge and readmission, the multiple organ dysfunction syndrome (MODS) scores on the initial admission, discharge and readmission were higher than the corresponding indices in the survivors. We conclude that the readmission rate was lower when intensivists participated in the discharge decision-making, and that APACHE and MODS scores on the first discharge and readmission were significant prognostic factors in respect of the readmitted patients. PMID- 15118989 TI - The evolution of lupus activity among patients with end-stage renal disease secondary to lupus nephritis. AB - The aim of this study was to evaluate the evolution of lupus activity in end stage renal disease (ESRD) patients due to lupus nephritis and to determine the long-term prognosis. We reviewed the clinical courses of 45 patients with ESRD due to systemic lupus erythematosus (SLE). We analyzed the course of SLE following the onset of ESRD, with special attention to the clinical and serological manifestations, survival time on dialysis, and renal transplantation outcome. Disease activity was measured using the SLE Disease Activity Index (SLEDAI). Of the 45 patients, 21 patients were being treated with hemodialysis (HD), 11 were undergoing peritoneal dialysis (PD), and 13 underwent transplantation. Duration of follow- up was 53 +/- 29 months. The SLEDAI score on commencement of renal replacement therapy was not significantly different among the 3 groups (HD: 4.2 +/- 4.2, PD: 4.3 +/- 2.3, Transplant: 3.2 +/- 1.9). However, disease activity scored by follow-up maximal SLEDAI during dialysis or transplantation showed a significant increase after peritoneal dialysis (HD: 5.0 +/- 3.6, PD: 7.4 +/- 3.7, Transplant: 2.2 +/- 1.7, p < 0.05). When the individual changes in the maximal SLEDAI score were considered, a significant increase was apparent after peritoneal dialysis (p < 0.05), but not after either hemodialysis or renal transplantation. There was no significant difference in cumulative survival rate, and also in technique or graft survival rates of the 3 groups. Among the variables tested, follow-up maximal SLEDAI score was the only significant factor associated with patient survival (odds ratio: 1.15, p < 0.05). The incidence (36% versus 19%) of high disease activity was greater, but not significantly, in the peritoneal dialysis group, as compared to the hemodialysis group. Clinical activity of SLE was apparent in 65% of patients in the first year of dialysis, but none showed any activity after the third year of dialysis. We found that although lupus disease activity declined after patients progressed to ESRD, lupus disease activity still affected patients' survival. An incremental increase in postdialysis lupus activity was not uncommon, especially during the first one year of dialysis. During the follow-up period, maximal SLEDAI score increased significantly after peritoneal dialysis. However, the long-term prognosis was not significantly different according to the treatment modality. PMID- 15118990 TI - Alcoholism prevalence and some related factors in Edirne, Turkey. AB - The aim of this research was to estimate the community prevalence of alcoholism and the potential risk factors that affect it in the Edirne provincial centre by using a scanning test. A cross-sectional study was carried out in the Edirne provincial centre. A sample population composed of 500 women and 200 men was selected randomly after the categorisation of the population according to ethnicity, age and sex. Through face-to-face interviews, data collection sheets, which were prepared to analyse potential factors affecting alcoholism frequency, were filled in by the sample population. The Michigan Alcoholism Scanning Test (MAST) was employed. According to MAST's normal grading, individuals with 5 or more points are evaluated as alcoholics. Accordingly, 8.2% of the sample population fit the definition of alcoholic. Alcoholism frequency was considerably higher in gypsies, the self-employed, smokers, and people with higher income. From logistic regression analysis alcoholism frequency was 12.4 times higher in men than in women, 3.2 times higher in gypsies than in others, 1.9 times higher in people who earned an income in the preceding week than in the unemployed, and 3.7 times higher in individuals who had smoked more than 100 cigarettes during their life or those who had smoked at least 1 cigarette for 3 months or for a longer period than in those who hadn't smoked any cigarettes. The prevalence of alcoholism in the Edirne provincial centre was similar to that in other countries in Europe. The most important finding was that alcohol consumption decreased in the unemployed, a finding that differs from that in other parts of the world. Gypsies, who differ in tradition, way of life, and job compared to the other strata of society, also suffered from higher alcohol consumption. This group usually consumed wine and generally did not eat while drinking. PMID- 15118991 TI - Increased serum level of p-selectin in patients with lichen planus. AB - Lichen planus (LP) is a common, pruritic and inflammatory disease of the skin, hair follicles and mucous membranes. Immunologic mechanisms, especially cell mediated immunity, play a major role in triggering the clinical expression of the disease. P-selectin is an adhesion molecule present within endothelial cells and mediates endothelial-leukocyte interactions. Therefore, it is considered that P selectin plays an important role in LP. The aim of our study is to research the relation between P-selectin and LP. Serum P-selectin levels were determined with the enzyme- linked immunosorbent sandwich assay method in sera from 40 LP patients and 40 healthy controls. The serum levels of P-selectin were statistically significantly higher in the patients than in healthy controls (p < 0.05), in female patients (39.32 +/- 11.34 pg/ml) than in male patients (31.93 +/ 9.83 pg/ml) (p < 0.01), and in the patients with eruptive form (40.27 +/- 9.32 pg/ml) than in those with the localised (32.83 +/- 9.93 pg/ml) and hypertrophic (31.72 +/- 8.39 pg/ml) forms (both p < 0.01). In conclusion, we found a meaningful relation between LP and serum P-selectin levels. PMID- 15118992 TI - Possible role of amyloid beta-(1-40)-BSA conjugates in transdifferentiation of lens epithelial cells. AB - We investigated whether amyloid beta(Abeta) aggregates have transforming growth factor beta- like cytokine activity and cause transdifferentiation of lens epithelial cells, leading to certain types of cataract. In order to mimic Abetaaggregates, Abeta-(1-40) was crosslinked to bovine serum albumin (BSA) with disuccinimidyl suberate according to a previously described procedure. When human lens epithelial B-3 (HLE B-3) cells were treated with the Abeta-(1-40)-BSA conjugates, we observed the translocation of Smad-3, as well as the induced mRNA levels of fibronectin (FN), collagen type I (Col I), smooth muscle actin (SMA) and matrix metalloproteinase-2 (MMP-2). In addition, we investigated the morphology of rat whole lens cultured for 5 days in the presence of Abeta-(1-40) BSA, and the immunohistochemical localizations of Abeta-(1-40)/amyloid precursor protein (APP) in human clinical tissues beneath the anterior capsules. In rat whole lens cultures, treatment with Abeta-(1-40)-BSA produced a transformed morphology that had multiple layers of lens epithelial cells. To compare the anterior capsules in anterior subcapsular cataracts with those in nuclear cataracts, immunohistochemical studies of Abeta/APP in human clinical tissues revealed that the predominant immunostaining of Abeta occurs in the anterior epithelial plaques, which likely produces the abnormal extracellular matrix. Thus, these findings suggest that Abeta aggregates in vivo are possibly involved in the regulatory process by which lens epithelial cells may transdifferentiate into fibroblast-like cells, as well as help understand the mechanisms which lead to certain types of cataractogenesis. PMID- 15118993 TI - Head injuries from falls in preschool children. AB - Falls are a leading cause of morbidity and mortality among children. We performed a retrospective analysis of pediatric patients under 7 years of age admitted to our department after a fall from January 1994 through December 1999 to describe the characteristics of fall-related head injury and to determine the clinical parameters influencing outcome. The patients were divided into two groups according to age: group I (babies and toddlers, 0-3 years) and group II (preschool children, 4-6 years). Falls were classified as low and high level. Sixty-eight cases were identified and falls accounted for 35.2% of head injuries. There were more boys than girls, and more low-level falls(LLF) than high-level falls (HLF), particularly in group I. Although more common in HLF, significant intracranial injuries were also sustained from LLF. Calvarial fractures were the most frequent type of head injury and were more common in LLF than HLF. Admission Glasgow Coma Scale score, types of head injury and hypoxia on admission were significantly correlated with Glasgow Outcome Scale score, but age, sex, extracranial injury and height of fall did not influence clinical outcome. From this study, we concluded that the height of fall should not limit the evaluation of patients and that aggressive management is mandatory to improve outcome even in patients with poor prognostic factors. PMID- 15118994 TI - The surgical effect of callosotomy in the treatment of intractable seizure. AB - We evaluated the surgical effects of the callosotomy, particularly with respect to the effect of callosotomy in some seizure types and the extent of surgery. Twenty-one patients with a minimum follow-up of two year were enrolled. The most significant effect of callosotomy was the complete suppression of the generalized seizures associated with drop attack in 12 of 21 patients and seizure reduction of more than 75% in 6 of 21 patients. The surgical effect on the partial seizures was very variable. Transient disconnection syndrome appeared in 4 patients after anterior callosotomy. Total callosotomy by staged operation significantly suppressed generalized seizures associated with drop attack without any disconnection syndrome. Our data show that callosotomy is quite a good approach to the surgical treatment of drop attacks accompanied by disabling generalized seizures. PMID- 15118995 TI - Plastic changes of motor network after constraint-induced movement therapy. AB - The effects of short-term constraint-induced movement (CIM) therapy on the activation of the motor network were investigated with functional magnetic resonance imaging (fMRI). Movement of the less-affected arms of five patients was restricted and intensive training of the affected upper limb was performed. Functional MRI was acquired before and after two-weeks of CIM therapy. All patients showed significant improvement of motor function in their paretic limbs after CIM therapy. For three patients, new activation in the contralateral motor/premotor cortices was observed after CIM therapy. Increased activation of the ipsilateral motor cortex and SMA was observed in the other patient. Our results demonstrated that plastic changes of the motor network occurred as a neural basis of the improvement subsequent to CIM therapy following brain injury. PMID- 15118996 TI - Increased incidence of carotid artery wall changes and associated variables in hemodialysis patients without symptomatic cardiovascular disease. AB - Cardiovascular disease (CVD) is still the major cause of the morbidity and mortality in hemodialysis (HD) patients. The characteristics of major arterial changes, atherosclerosis and related risk factors in HD patients remain unclear. We aimed to evaluate the atherosclerotic process in asymptomatic HD patients and healthy volunteers, and to determine the association between the risk factor(s) and the atherosclerotic process in these groups. 92 HD patients (female: 43, male: 49) and 62 age and sex matched healthy volunteers (female: 27, male: 35) were enrolled in this study. Diabetics, smokers, and patients with symptomatic CVD were excluded. The right and left carotid intima-media thicknesses (CIMTs) were measured and plaque structures were studied by B-mode ultrasound. The mean CIMT in patients and control group were 0.79 +/- 0.16 mm and 0.54 +/- 0.09 mm, respectively. Mean CIMT in HD patients was thicker (p < 0.001) and the presence ratio of plaque was higher in patients group (n=38, %61.2 vs n=9, %17.3) (p < 0.001). Calcified type of plaque was more frequent in HD patients than control group. Age (r=0.48, p < 0.001), left ventricular mass (r=0.42, p < 0.05), and homocysteine (r=0.46, p < 0.01), mean hematocrit (r=-0.36, p < 0.05), plasma CRP (r=0.50, p < 0.001), ESR (r=0.43, p < 0.01) and albumin (r= -0.34, p < 0.05) levels were correlated with the CIMT measurements and plaque presence, significantly. -CIMT as an atherosclerotic process indicator is thicker in asymptomatic HD patients than healthy subjects. We concluded that in addition to various classical risk factors, uremic environment may also contribute to acceleration of the atherosclerotic process. PMID- 15118997 TI - Assessment of extent of myocardial ischemia in patients with non-ST elevation acute coronary syndrome using serum B-type natriuretic peptide level. AB - Since B-type natriuretic peptide (BNP) concentration has been shown by recent studies to be elevated in patients presenting acute coronary syndrome (ACS) even in the absence of overt heart failure, other mechanisms for elevating plasma BNP (p-BNP) concentrations may be suggested to exist. We have studied the correlation between p-BNP level and the extent of myocardial ischemia (EMI) in non-ST elevation (NSTE) ACS and evaluated the BNP level as an objective marker of EMI. In 204 patients with NSTE ACS, we estimated the EMI by the echocardiographic wall motion score index (WMSI) and the coronary angiographic Gensini score. As the positive control group, 44 patients with stable angina were enrolled into the study. We compared their initial p-BNP levels with WMSI and the Gensini score. Additionally, peak troponin-T level was compared with p-BNP level in NSTE myocardial infarction (MI) patients. Using the multiple regression analysis, adjustments for age, left ventricle (LV) wall stress, LV mass amount and ejection fraction (EF) were made. Patients with LVEF < 45% or age > 75 years or underlying diseases that could elevate BNP levels were excluded from the study. P-BNP level was increased in NSTE ACS patients compared with stable angina patients (133.9 +/ 87.4 vs. 12.2 +/- 9.2 pg/ml, p < 0.05). P-BNP levels were found to correlate with WMSI and the Gensini score in unstable angina (r=0.519, p < 0.01; r=0.680, p < 0.01) and NSTEMI (r=0.716, p < 0.01; r=0.684, p < 0.01) patients, respectively. Additionally, p-BNP levels correlated with the peak troponin-T level in patients with NSTEMI (r=0.700, p < 0.01). P-BNP level might be a useful marker in the assessment of EMI. PMID- 15118998 TI - Differential thioredoxin reductase activity from human normal hepatic and hepatoma cell lines. AB - Thioredoxin reductase (TrxR), a component of the thioredoxin system, including thioredoxin (Trx) and NADPH, catalyzes the transfer of electrons from NADPH to Trx, acts as a reductant of disulfide-containing proteins and participates in the defense system against oxidative stresses. In this study, the regulation pattern of TrxR in the presence of various stressful reagents was compared between Chang (human normal hepatic cell) and HepG2 (human hepatoma cell) cell lines. Aluminum chloride (0.5 mM) and zinc chloride (0.5 mM) enhanced the TrxR activity in the Chang cell line to a higher degree than in the HepG2 cell line, but cupric chloride (0.2 mM) and cadmium chloride (0.1 mM) enhanced the TrxR activity in the HepG2 cell line to a greater degree. The TrxR activities in both Chang and HepG2 cell lines were similarly induced by treatment with sodium selenite (0.02 mM) and menadione (0.5 and 1.0 mM). Lipopolysaccharide (2 micro g/m1) increased the TrxR activity upto 4.02- and 2.2-fold in the Chang and HepG2 cell lines, respectively, in time-dependent manners. Hydrogen peroxide (5 mM) markedly enhanced the TrxR activity in the HepG2 cell line, but not in the Chang cell line. NO-generating sodium nitroprusside (3.0 and 6.0 mM) induced TrxR activities in both human liver cell lines. The TrxR activity was also induced in human liver cells under limited growth conditions by serum deprivation. These results imply that the TrxR activities in normal hepatic and hepatoma cell lines are subject to different regulatory responses to various stresses. PMID- 15118999 TI - Gestational age--the most important factor of neonatal ponderal index. AB - Ponderal index (fetal weight in grams X 100 / (fetal length in centimeters)3) (PI) is one of the anthropometric methods used to diagnose impaired fetal growth. Irrespective of the infant's position on the growth-weight-for-gestational age charts, PI is low in malnourished infants and high in obese ones. As fetal growth is affected by ethnicity, geographic location and socioeconomic status, we developed standards for neonatal PI, and assessed the effects of gestational age, sex and maternal parity. Data on 5798 newborns from singleton pregnancies born in the Department of Gynecology and Obstetrics, Split University Hospital, were retrospectively analyzed. Over a 15-month period in 2000/2001, 5596 newborns from 24 to 42 weeks of gestation were born. The other 202 newborns, born from 24 to 34 weeks of gestation in the ten year period, 1990-1999, were added because of the small number of preterm infants; ensuring a minimum of 30 to fill up at least infants in each gestational week. All mothers were of Caucasian origin. Stillbirths and fetuses with congenital malformations were excluded. The 10th, 50th and 90th percentiles, mean values with standard deviation of PI and the 10th, 50th, and 90th percentiles of birth weight and birth length are presented separately at weekly intervals. PI showed linear correlation with gestational age from 24 to 39 weeks, after witch the data plateaued. Sex and parity had no impact on PI in infants born between 24 and 37 weeks. Analysis of variance revealed PI to be significantly higher in female than in male newborns, and in multiparous than in nulliparous infants after 37 weeks of gestation. In conclusion, gestational age is the most important factor of neonatal PI. The effects of sex and parity on PI should only be considered in term neonates. PMID- 15119000 TI - Comparison of perioperative complications between reconstructive pelvic surgery and general gynecologic surgery. AB - The purpose of this study was to compare the perioperative complication rates of reconstructive pelvic surgery and general gynecologic surgery, and to identify the predictive risk factors for perioperative complications in reconstructive pelvic surgery. The medical records of 148 reconstructive pelvic surgery patients and 146 general gynecologic surgery patients were reviewed, and the types of complications, along with their rates and predictive risk factors were examined. The statistical analysis included descriptive statistics and logistic regression. There was no difference in the type of complications between reconstructive pelvic surgery and general gynecologic surgery. The prevalences of perioperative complications were 34.4% in the reconstructive pelvic surgery group and 26.7% in the general gynecologic surgery group. Intraoperative blood loss (p= 0.006) and the duration of surgery (p=0.014) were independent risk factors for perioperative complications in the reconstructive pelvic surgery group. The perioperative complication rates for the patients undergoing reconstructive pelvic surgery were not higher than those of the patients undergoing general gynecologic surgery, even though more procedures were performed and a longer duration of surgery was needed in the former cases. Since the duration of surgery and the amount of blood loss are the major factors affecting the complication rate, decreasing these two factors would be the key to improving the outcomes of patients undergoing reconstructive pelvic surgery PMID- 15119001 TI - The predictive values of various parameters in the diagnosis of stress urinary incontinence. AB - The Maximum Urethral Closure Pressure (MUCP) and Functional Urethral Length (FUL) are significant parameters of the Urethral Pressure Profile (UPP), while the Q tip angle and Bladder Neck Descent (BND) are the significant parameters of urethral hypermobility. We performed a study to evaluate the effects and predictive values of each of these parameters in the diagnosis of Stress Urinary Incontinence (SUI). A retrospective study was done involving 90 SUI patients and 38 non-SUI patients who underwent urodynamic study, Q-tip test and perineal ultrasound at Yonsei Medical Center between January, 1999 and February, 2002. There was no statistical difference between the SUI and non-SUI groups in terms of mean age, delivery history, menopausal age and body mass index. While the FUL and Q-tip angle showed significant differences (33.18 +/- 19.55 vs 33.12 +/- 13.37 mm, p=0.002; 65.94 +/- 21.69 vs 56.45 +/- 26.53 degrees C, p=0.02,respectively) neither the MUCP nor the BND showed any significant difference between the two groups (60.06 +/- 29.92 vs 48.97 +/- 42.95 cmH2O, p > 0.05; 1.09 +/- 0.75 vs 0.85 +/- 0.76 cm, p > 0.05; 0.71 +/- 0.80 vs 0.53 +/- 0.72 cm, p > 0.05). The odds ratios for the FUL and Q-tip angle were 1.038 (1.014, 1.061) and 1.017 (1.001, 1.033), respectively. The FUL and Q-tip angle had cut off values of 1.36 cm (sensitivity: 68.8%, specificity : 54.1%, PPV : 73.8%, NPV : 48.1%) and 20.47 degrees C (sensitivity : 93.3%, specificity : 18.17%, PPV : 68.2%, NPV : 60%), respectively, in the diagnosis of SUI. The area under the curve (AUC) of the FUL and Q-tip angle were on average 0.625 (p=0.0016) and 0.575 (p=0.0012), respectively. Both the FUL and Q-tip angle showed a significant difference between SUI patients and the normal group. However, their value as a diagnostic tool was trivial, and since their sensitivity, specificity, positive predictive value and negative predictive value showed inconsistent results at each cut-off value, it would be difficult to apply them to clinical use. A further study is required to set-up standard diagnostic values of these variables for clinical use. PMID- 15119002 TI - Wear of retrieved UHMWPE hip liners. AB - After the gamma-irradiation sterilization, the most widely used orthopaedic grade polymer bearing liner material for the total joint replacement, ultra-high molecular weight polyethylene (UHMWPE), degrades through the progressive in vivo oxidation. The oxidative degradation makes UHMWPE brittle and leads to reduction of its mechanical properties. In this study, the effect of the in vivo post irradiation ageing time on the wear of UHMWPE was investigated. Twelve retrieved polyethylene hip liners implanted for 3-16 years and then stored in the air for 1.5-8 years were used. Two types of the pin-on-disk wear testing were conducted. The uni-directional repeat pass rotating and the linear reciprocating wear testing were done with stainless steel disks against stationary polyethylene pins under 4MPa at 1Hz with bovine serum lubrication. Wear of the retrieved polyethylene hip liners does not have significant correlation with the in vivo or total ageing time. The linear reciprocal sliding motion generated a more pronounced wear than the uni-directional repeat pass sliding motion. This indicates that the kinematic motion significantly affects the wear of aged UHMWPE, having a brittle, white band region. PMID- 15119003 TI - Tattoo and personality traits in Croatian veterans. AB - To examine whether tattooed patients, treated for posttraumatic stress disorder (PTSD) caused by war at the Ward for Psycho-trauma of the Clinical Hospital Osijek, differ from non-tattooed patients by certain personality traits. The study was conducted on one hundred Croatian veterans who were divided into two groups with respect to the presence/ absence of tattoo. To assess the symptoms of PTSD, the Clinical Administered PTSD Scale (CAPS-2) was used for all subjects. To assess personality traits the following psychology tests were applied: Purdue non verbal IQ test, Minnesota Multiphasic Personality Inventory (MMPI-1), and Eysenck's Personality Questionnaire (EPQ/A and EPQ/IVE). With respect to the examined pre-traumatic variables and PTSD symptoms, the two groups manifested no differences. The non-tattooed group achieved higher scores on the IQ test (IQ=100) than the tattooed group (IQ=95). EPQ test showed results either above or below the norms on all scales that were applied. The tattooed group demonstrated significantly higher levels of impulsiveness, adventurism, empathy and neuroticism than the non-tattooed one (p < 0.05). In the group of 100 Croatian veterans treated for PTSD, 33 had tattoos and 67 did not. The results indicated more impulsiveness, adventurism / risk behavior, empathy and neuroticism in the tattooed group than in the non-tattooed group, while there was no significant difference in the intensity of the PTSD symptoms. PMID- 15119004 TI - Expression of down stream molecules of RET (p-ERK, p-p38 MAPK, p-JNK and p-AKT) in papillary thyroid carcinomas. AB - To evaluate the roles of 4 putative downstream molecules (ERK, p38 MAPK, JNK and AKT) of the RET signal pathway in the tumorigenesis of papillary carcinomas, the expression patterns of RET and phosphorylated forms of ERK, p38 MAPK, JNK and AKT were evaluated in 115 cases of papillary thyroid carcinomas by 3 mm-core tissue microarray based immunohistochemical staining. The prevalence of RET protein expression was 62.6%. No distinct expression of p-ERK and p-p38 MAPK was demonstrated in tumor cells of papillary carcinomas. All papillary carcinomas except 5 cases expressed nuclear p-JNK and p-JNK expression was increased in tumors compared with paired normal tissues (p < 0.05). There was no difference in the p-JNK expression between RET protein-positive and RET protein-negative papillary carcinomas (p > 0.05). Unequivocal nuclear staining for p-AKT was demonstrated only in 10 cases of papillary carcinomas, and all of them showed focal staining. Our results showing constitutive expression of p-JNK in most cases of surgically excised papillary thyroid carcinomas irrespective of RET protein expression status suggest that JNK activation may play a role in the tumorigenesis or survival of sporadic papillary thyroid carcinoma. PMID- 15119005 TI - Differential effect of vitamin K and vitamin D supplementation on bone mass in young rats fed normal or low calcium diet. AB - The purpose of this study was to clarify the differential effect of vitamin K and vitamin D supplementation on bone mass in young rats fed a normal or low calcium diet. Ninety female Sprague-Dawley rats, 6 weeks of age, were randomized by stratified weight method into nine groups with 10 rats in each group: baseline control, and 0.5% (normal) or 0.1% (low) calcium diet, either alone, or with vitamin K (30 mg/100g, food intake), vitamin D (25 micro g/100 g, food intake), or vitamin K + vitamin D. After 10 weeks of feeding, bone histomorphometric analyses were performed on cortical bone of the tibial shaft and cancellous bone of the proximal tibia. Vitamin K supplementation increased the maturation-related cancellous bone gain and retarded the reduction in the maturation-related cortical bone gain in rats fed a low calcium diet, and increased the maturation related cortical bone gain in rats fed a normal calcium diet. Vitamin D supplementation reduced the maturation-related cancellous bone gain, prevented the reduction in periosteal bone gain, and enhanced the enlargement of the marrow cavity, with no significant effect on the reduction in the maturation-related cortical bone gain in rats fed a low calcium diet, and increased the maturation- related cancellous and cortical bone gains with increased periosteal bone gain in rats fed a normal calcium diet. An additive effect of vitamin K and vitamin D on the maturation- related cortical bone gain was found in rats fed a normal calcium diet. This study shows the differential effects of vitamin K and vitamin D supplementation on cancellous and cortical bone mass in young rats fed a normal or low calcium diet, as well as the additive effect on cortical bone under calcium sufficient condition. PMID- 15119006 TI - Occupational asthma due to azodicarbonamide. AB - Azodicarbonamide is a low molecular weight foaming agent for plastics and rubbers. Azodicarbonamide can elicit acute and chronic health related problems due to its potential for pulmonary and cutaneous sensitization. Some cases of occupational asthma associated with exposure to azodicarbonamide have been reported, of which only a few cases were confirmed by specific inhalation challenges. Here, the first case of occupational asthma due to azodicarbonamide in Korea, in which the diagnosis was confirmed by specific inhalation challenge, is reported. PMID- 15119007 TI - Isolated extramedullary relapse of acute myelogenous leukemia as a uterine granulocytic sarcoma in an allogeneic hematopoietic stem cell transplantation recipient. AB - We report an unusual case of acute myelogenous leukemia in a patient who showed an extramedullary relapse in her uterus, without bone marrow recurrence, two years after an allogeneic bone marrow transplant. She complained of irregular vaginal spotting, and magnetic resonance imaging demonstrated a uterine mass. A biopsy revealed a massive infiltration of immature myeloid cells. A variable number of tandem repeats (VNTR) based on an examination of peripheral blood cells showed full donor chimerism. After receiving chemotherapy, her uterine mass had completely resolved. She has remained in complete remission for more than 6 months. This case suggests that physicians should be aware of the possibility of a uterine relapse in female bone marrow transplant recipients with acute myelogenous leukemia. PMID- 15119008 TI - First Turkish patient with floating harbor syndrome with additional findings: cryptorchidim and microcephaly. AB - We report the first Turkish patient with Floating Harbor Syndrome (FHS). The 12 year old male patient exhibited classical dysmorphic features of FHS, mental retardation, celiac disease and additional undescribed findings: microcephaly and cryptorchidism. PMID- 15119009 TI - Phenytoin-induced gingival overgrowth in un-cooperated epilepsy patients. AB - Phenytoin-induced gingival overgrowth is a well-known and frequently reported gingival lesion, which was first detected in 1939. However, there are conflicts in the literature about the agents which affect the severity of the lesion. Un cooperative dental patients are one of the most unsuccessfully treated periodontal patient groups because of the difficulty in maintaining their oral hygiene. This case report consists of two cases with the same characteristics: phenytoin usage, comprehension and speech defects and poor oral hygiene, but each case differs in the duration of the phenytoin therapy. Both of the cases received scaling, root planning and a gingivectomy. PMID- 15119010 TI - Graves' disease associated with Klinefelter's syndrome. AB - Klinefelter's syndrome is one of the most common forms of primary hypogonadism and infertility in males. It is characterized by small and firm testes, gynecomastia, azoospermia, and an elevated gonadotropin level. The frequencies of diabetes mellitus, breast cancer, and germ cell neoplasia increases in Klinefelter's syndrome. We report upon a 35 year-old male patient with Graves' disease in association with Klinefelter's syndrome; as confirmed by chromosome analysis. The patient is being treated with antithyroid medication for Graves' disease and by testosterone replacement for Klinefelter's syndrome. PMID- 15119011 TI - Primary malignant melanoma of the cervical spinal nerve root. AB - The authors report on a case of primary malignant melanoma of the 7th cervical spinal nerve root in a 45-year-old woman. Neuro-radiological features of this extra-dural mass were suggestive of a nerve sheath tumor. The lesion underwent total gross resection through the anterolateral approach. The patient's postoperative course was uneventful. Histopathological investigation confirmed malignant melanoma. There was no evidence of tumor recurrence or other melanotic lesions on regular follow-up examinations until the postoperative eighth month. When treating a common, benign-looking lesion of the cervical spinal nerve root, surgeons should be aware of the potential to encounter such a malignant tumor. PMID- 15119012 TI - A new case of fatal pulmonary thromboembolism associated with prolonged sitting at computer in Korea. AB - A newly reported, variant form of venous thromboembolism (VTE), named as "eThrombosis", occurred in a patient who maintained a prolonged sitting position at a computer. The patient fortunately recovered from the disease through early diagnosis and treatment. Recently however, the author experienced a new case of a 24-year-old Korean man who died due to "eThrombosis" after playing an Internet computer game continuously for about 80 hours. Based on this case, the author would like to warn heavy computer users of the risk of "eThrombosis", as they might also be at risk. Sitting for a long time at the computer results in an immobility that is a new and large risk factor of life-threatening VTE. PMID- 15119013 TI - A case of adult-onset centronuclear myopathy. AB - Centronuclear myopathy (CNM) is a rare congenital myopathy that is characterized by centrally placed nuclei in the muscle fibers. Based on the time of onset and the mode of inheritance, CNM can be divided into three distinct forms: the severe neonatal form, the childhood onset form, and the adult onset form. This paper describes the case of a female patient with CNM, in whom the disease manifested itself in the fifth decade of life, without any prior family history of such disorders. To the best of our knowledge, this is a rare case of late adult-onset CNM. PMID- 15119014 TI - Intestinal obstruction due to a mesenteric cyst. AB - Mesenteric cysts are rarely thought of, may be difficult to diagnose, and are usually asymptomatic except when complicated. Intestinal obstruction is a rarely reported complication of these cysts. A case of mesenteric cyst that was causing obstruction of the large bowel is presented, along with a review of the literature. PMID- 15119015 TI - Cloning of TLR3 isoform. AB - Toll-like receptor (TLR) 3 is a member of the TLR family that confers innate immunity by recognizing viral pathogens. Herein, we report that the TLR3 isoform is expressed on human primary cells and cell lines. This isoform has 2,520 bp cDNAs compared to the 2,712 bp of full cDNA, is produced by deletion of an intron like sequence within exon 4 and is co-expressed with wild type TLR3 in primary human astrocytes and glioblastoma cell lines. This finding suggests the TLR3 isoform in astrocytes may have a different immunological role for binding ligands during the immune response in brain. PMID- 15119016 TI - Proceedings of the 8th International Conference on Primary Therapy of Early Breast Cancer. St. Gallen, Switzerland, 12-15 March 2003. PMID- 15119017 TI - The EAMG position on the regulation of existing products for treatment with special reference to named patient products (NPPs). AB - Allergen products have a long history in both diagnosis and management of allergic disease. It is emphasized that the availability of named patient products presents a valuable and indispensable option for the effective and safe treatment of patients suffering from IgE-mediated allergic diseases. A regulatory climate should be achieved in which these products can survive until a safe and effective alternative is borne out. Within the context of the paper the following definition is used: A named patient product or NPP is an allergen product, prepared with a specific reference number in accordance with a prescription for an individual patient, identified by the name of this patient and the reference number, and delivered after control of consistency with previous treatments. It is stressed that the standards of quality that hold for registered products are also applicable for named patient preparations which, for various reasons, can not be filed for registration. The paper frequently refers to a Position Paper that has recently been adopted by the European Allergen Manufacturers Group, the EAMG. The presentation breaks a lance for a concerted approach of the NPP-concept by striving at harmonization of legal, regulatory, manufacturing, and distribution conditions. PMID- 15119018 TI - Specific immunotherapy in the U.S.A.: general concept and recent initiatives. PMID- 15119019 TI - Sample size considerations for establishing clinical bioequivalence of allergen formulations. AB - Bioequivalence of formulations must be established by proving that the differences between the formulations are within a specified interval according to Equation 1, the Interval Hypothesis. Explicit estimates of sample size determined from Equation 8 and listed in Table 1 are qualitatively larger than those that would be determined from Equation 2, the Hypothesis of No Difference. Equation 8 was derived from the TOST procedure; other valid methods should yield comparable results. In any context, this discussion has illustrated that the failure to demonstrate a difference is not sufficient to demonstrate equivalence, and that a properly powered equivalence study of allergen formulations will generally demand many more than four study subjects. PMID- 15119020 TI - Future aspects of regulation. PMID- 15119021 TI - The manufacturers' perspective: regulation of allergen products in European countries. PMID- 15119022 TI - Overview: biological standardization in Europe and the USA. PMID- 15119023 TI - The CREATE project: a new beginning of allergen standardization based on mass units of major allergens. PMID- 15119024 TI - Mediator release assays based on human and murine IgE: potential and limitations in allergen standardization. PMID- 15119025 TI - In vitro and in vivo characterization of hazelnut skin prick test extracts. AB - RATIONALE: Hazelnut allergy ranks among the most frequently observed food allergies. Clinical symptoms range from the oral allergy syndrome to life threatening anaphylaxis. Diagnosis of hazelnut allergy partially relies on in vivo testing by means of skin prick testing (SPT). The aim of this study was to characterize hazelnut SPT extracts both in vitro and in vivo. METHODS: Hazelnut SPT extracts were investigated for protein concentration and composition. The major hazelnut allergen Cor a 1, lipid transfer protein (LTP) and thaumatin-like protein (TLP) were monitored by competitive RIA and immunoblotting. SPT extracts (n = 6) were analyzed for skin reactivity and the correlation between the SPT extract protein concentration and the mean skin reactivity (HEIC) was determined in a group of hazelnut-allergic patients (n = 30). For one SPT extract, the threshold level for Cor a 1 was determined in Cor a 1-monosensitized patients (n = 5). RESULTS: Protein concentrations ranged from 0.2-14 mg/ml. Although some proteins were present in most extracts (bands at 10, 22-28, 32 and around 48 kDa), clear differences in composition were observed (both intra- and inter variability). The concentration of the major hazelnut allergen Cor a 1 differed up to a factor 50 (0.6-32 micrograms/ml). LTP was virtually absent in 3/9 SPT extracts and variable quantities of TLP were detected by immunoblotting. Some patients (6/30) had a false-negative SPT with 3/6 SPT extracts. There was a clear correlation between the protein concentration and the mean HEIC (RPearson = 0.87). The threshold level for Cor a 1 was +/- 3.2 ng/ml as assessed with one of the products investigated. CONCLUSIONS: Heterogeneous protein concentration/composition of SPT extracts results in variable skin test responses. The absence of potentially severe allergens like LTP may lead to false negative SPT results that jeopardize a patient's safety. From these results it can be concluded that there is a strong need for standardization of products for SPT. PMID- 15119026 TI - Immunochemical and biological quantification of peanut extract. AB - Biological standardization of allergen extracts is one of the steps in the characterization of an extract. The gold standard for determination of biological potency is the skin prick test, but histamine release (HR) has been used as a convenient ex vivo method for analyzing a large number of samples. We describe the use of rabbit basophils as a tool in biological standardization. Using peanut as a model allergen, it is described how rabbits immunized for production of antiserum may become sensitized and their basophils used for histamine release experiments. It is also possible to use rabbit antiserum to passively sensitize basophils derived from naive rabbits, but the sensitivity of this method is so far 100-1000 times lower than the direct histamine release. The rabbit histamine release results are compared to an ELISA developed by means of the same antisera and by passive sensitization of human basophils using serum from a strongly sensitized peanut-allergic patient. The overall sensitivity of the methods were ELISA > HR-human cells > HR-sensitized rabbit cells > HR-passively sensitized rabbit cells. The use of rabbit basophils for biological standardizations will allow for the use of rabbit antisera. PMID- 15119027 TI - Latex: a new target for standardization. PMID- 15119028 TI - Criteria for clinical efficacy--readout and monitoring of clinical studies. PMID- 15119029 TI - Oral, nasal and sublingual immunotherapy: do they work, are they safe? PMID- 15119030 TI - Peptide-based immunotherapy: new developments. PMID- 15119031 TI - Preventive aspects of specific immunotherapy. PMID- 15119032 TI - Developing a manufacturing process and analyses for a recombinant protein drug. PMID- 15119033 TI - Engineering and validation of recombinant proteins for allergen vaccines. PMID- 15119034 TI - Mechanisms of immunotherapy with allergoids: lessons for the development of recombinant allergens with reduced IgE-binding activity. PMID- 15119035 TI - Utility of pure allergen components in the continuing development of in vitro diagnostics for inhalant and food allergies. PMID- 15119036 TI - Comparison of natural grass pollen allergens and their recombinant counterparts expressed in bacterial and mammalian systems. PMID- 15119037 TI - Expression of allergens in E. coli and plants--benefits and drawbacks. AB - Recombinant allergens are quickly becoming the reagents of choice for diagnosis and therapy of type I allergic diseases. Consequently, the different methods for the production of recombinant proteins that are available today are of great interest to allergologists. Without doubt, bacterial expression will continue to play a pivotal role. In addition, plant-based expression systems will be needed to overcome problems inherent in the E. coli systems and to allow the production of glycoallergens or allergens of more complex folding. PMID- 15119038 TI - Approaches based on mutated or modified recombinant grass pollen allergens--in vivo evaluation of the constructs. PMID- 15119039 TI - Recombinant food allergens in the diagnosis of pollen-related food allergy. PMID- 15119040 TI - Hypoallergenic variants of the Parietaria judaica major allergen Par j 1: in vitro and in vivo evaluation. PMID- 15119041 TI - Recombinant mite allergens: validation strategies and clinical trials. PMID- 15119042 TI - Mechanisms of specific immunotherapy: current knowledge. AB - Induction of specific unresponsiveness (tolerance) in peripheral T cells by IL-10 and/or TGF-beta and recovery by cytokines from the tissue microenvironment represent two key steps in specific immunotherapy of allergy and natural exposure to allergens in healthy individuals. IL-10 and TGF-beta elicit tolerance in T cells and thereby control the suppression and development of antigen-specific immunity. Both cytokines also play an important role on the generation of a non inflammatory IgG4 and IgA type of allergen--specific antibodies during the course of specific immunotherapy. Histamine plays an important role in upper and lower airway inflammation. In addition to its well-characterized effects in the acute inflammatory and allergic responses, histamine regulates several aspects of antigen-specific immune response development. Histamine affects the maturation of dendritic cells and alters their T cell polarizing capacity. Histamine regulates antigen specific Th1 and Th2 cells as well as related antibody isotype responses. Histamine and four different known histamine receptors (HR) display a very complex system and their expression changes according to the stage of cell differentiation as well as microenvironmental influences. PMID- 15119043 TI - Allergic rhinitis and its impact on asthma (ARIA)--what does it mean for the future of SIT? PMID- 15119045 TI - Alternative approaches to immunotherapy of allergy. AB - Alternative therapies for allergic diseases may be prescribed by physicians or provided by other practitioners or selected by patients without resorting to consultation with a health advisor. The most popular alternative therapies include dietary manipulation and exaggerated avoidance of allergens. Established medical therapies include herbs (Western and Asian), homeopathy and acupuncture; many publications suggest they are of value in allergic disease management. However, most studies lack sufficient power, do not have adequate controls, fail to duplicate standard clinical practices, or utilize a heterogeneous group of subjects. Careful analysis suggest that popular alternative therapies might offer a minimal benefit over placebos, but further studies are required for more rigorous evaluation of outcomes. Numerous esoteric or non-specific methods of treating allergic diseases are also available which may be promoted by enthusiastic practitioners or supported by satisfied patients. The majority of such therapies will never be scientifically evaluated, and the results of such modalities will be difficult to separate out from those of true placebo treatments. It is evident that while the "prose" of scientifically established therapies is appealing to a declining number of patients, the "poetry" of complementary therapies resonates with an increasing majority of the public who believe in the magic of alternative medicine. PMID- 15119044 TI - New perspectives for the treatment of food allergy (peanut). PMID- 15119046 TI - Parallels between parasite infections and specific immunotherapy. PMID- 15119047 TI - Anti-IgE therapy combined with specific immunotherapy: pro. PMID- 15119048 TI - Anti-IgE therapy combined with SIT: contra. PMID- 15119049 TI - Strategies for the development of safe and effective DNA vaccines for allergy treatment. AB - During the past ten years, a great number of studies have demonstrated that injection of plasmid DNA coding for certain genes results in the induction of humoral and cellular immune responses against the respective gene product. The features of DNA vaccines enable a broad range of applications, including the induction of protective immunity against viral, bacterial, and parasitic infections, and open up new perspectives for the treatment of cancer. Furthermore, based on their Th1-promoting properties, DNA vaccines also turned out to balance Th2-mediated immune reactions, a quality which renders them a promising alternative for immunotherapy against allergy. Their unique immunological properties offer new possibilities for the development of vaccines, which do not cause anaphylactic side effects, a major drawback of specific immunotherapy (SIT). In this review, we present approaches to avoid the translation of native allergenic determinants, thus preventing release of allergy mediators stimulated by crosslinking of pre-existing or vaccine-induced IgE antibodies on mast cells. Three approaches are described, which fulfill these requirements: (i) cutting the allergen gene into overlapping fragments, which lack any antigenic determinant of the native allergen, but display the original repertoire of T cell epitopes, (ii) using hypoallergenic derivatives or (iii) fusing the allergen with ubiquitin, thus fragmenting the antigen and destroying its native structure. The presented experiments demonstrate that DNA vaccines are suitable to balance an allergic response in a protective as well as a therapeutic experimental design, thus demonstrating their potential for allergy treatment. In addition to conventional plasmid DNA vaccines, aspects and perspectives of replicon-based DNA vaccines will be discussed. PMID- 15119050 TI - Protection against infectious diseases by DNA-based vaccination strategies- lessons for allergy treatment? PMID- 15119051 TI - Progress in the development of new methods of immunotherapy: potential application of immunostimulatory DNA-conjugated to allergens for treatment of allergic respiratory conditions. AB - Allergen immunotherapy was first introduced in the early part of the twentieth century. It is widely practiced despite having specific limitations. Considerable effort has been devoted to developing new modified allergens that, compared with conventional allergen immunotherapy, improve efficacy, decrease the time required to achieve effect, reduce inconvenience, and enhance safety. Increased understanding of allergic respiratory inflammation has led to the development of therapeutic modalities that potentially arrest the disease process in asthma or allergic rhinitis. This paper addresses an adjuvant approach in which highly active immunostimulatory phosphorothioate oligodeoxyribonucleotide sequence (i.e. immunostimulatory DNA) are conjugated to the principal allergenic moiety of a relevant aeroallergen. We have recently completed the first human safety studies in patients with allergic rhinitis with Amb a 1-immuno-stimulatory oligonucleotide conjugate (AIC) --a novel therapeutic vaccine comprised of Amb a 1, the principal allergenic protein of ragweed, conjugated specific immunostimulatory oligonucleotides (ISS). The results demonstrate that AIC was several hundred-fold less reactive than a standardized ragweed extract when evaluated by quantitative intradermal skin titration methodology. Furthermore, AIC reduced histamine release from basophils to a similar degree. The DNA vaccine induced IgG antibody production in treated patients. AIC compared with standardized aqueous ragwood exhibited fewer local reactions on skin testing, a finding that suggests that AIC offers the potential for an improved safety profile for immunotherapy. Additional trials to further evaluate the safety, immunologic effect, and therapeutic efficacy of AIC for ragwood-induced allergic rhinitis and asthma are ongoing. PMID- 15119052 TI - History and future of allergen standardization and of the Paul-Ehrlich-Seminar. PMID- 15119053 TI - Bacterial resistance: how to detect three types. AB - Antimicrobial susceptibility testing in the clinical laboratory is becoming more complex. We can no longer be concerned with simply determining accurate susceptibilities. Microbiologists must now possess knowledge of bacterial resistance mechanisms and implement procedures to reliably detect them. The traditional susceptibility test will most likely need to be supplemented with testing methods and software that allow for phenotypic identification of resistance mechanisms. Resistance mechanisms can be present in apparently susceptible bacterial populations. Accurate identification of these mechanisms will help to control emergence of new resistance by encouraging use of the most appropriate antibiotics. PMID- 15119054 TI - Follow a "world-class service" blueprint. Cleveland Clinic Foundation wins the MLO Medical Laboratory of the Year 2004 Award. PMID- 15119055 TI - Gen-Probe's Henry Nordhoff keys in on future trends. Interview by Dottie Dunham. PMID- 15119056 TI - Biochemical markers of bone turnover and their utility in osteoporosis. PMID- 15119057 TI - Another privacy issue: slide disposal. PMID- 15119058 TI - OSHA on phlebotomy disposal. PMID- 15119059 TI - Connectivity. Getting staff to use patient safety tools. AB - Technology is often hailed as the panacea for patient safety problems. But technology is only as good as the people who use it, the programs designed to train users, and the human systems in place to implement new equipment. PMID- 15119060 TI - ED outsourcing: is it good for patient care? AB - A debate rages over outsourcing physician recruitment and staffing of emergency departments. See what's involved for your board should you face a decision about whether or not to go with a contract management group. PMID- 15119061 TI - Conflict of interest and governance: new approaches for a new environment. PMID- 15119062 TI - Getting to know you. AB - When the board makes an effort to listen to the needs and concerns of physicians and shows its appreciation for a difficult job done well, the relationship between the two groups improves dramatically. PMID- 15119063 TI - Journey to excellence: a systemwide culture change begins with the board. AB - When a hospital system decided its goal should be becoming "best in the nation," it determined that the board should lead a systemwide culture change. It started with an innovative "shadowing" program. PMID- 15119064 TI - Patient safety. A prescription for certainty: a new wrong site surgery protocol. PMID- 15119065 TI - Insecticide treated nets: impact on vector populations and relevance of initial intensity of transmission and pyrethroid resistance. AB - Insecticide treated bednets locate a deposit of a quick-acting insecticide of low human toxicity between a sleeper and host-seeking mosquitoes. Thus a chemical barrier is added to the often incomplete physical barrier provided by the net. Treated nets may be considered as mosquito traps baited by the odour of the sleeper. Trials in Assam, Tanzania and elsewhere have shown that when a whole community is provided with treated nets, so many mosquitoes of anthropophilic species are killed by contact with the nets that the density and/or sporozoite rate of the vector population is reduced. In order to gain this "mass" or community effect, in addition to widespread personal protection, and thus to achieve the full potential of the treated net method, a high per cent coverage of the community is needed. This suggests that organised free provision of treated nets, comparable to a house spraying programme, is likely to be more cost effective than trying to market nets and insecticide to very poor rural people. In areas with high malaria transmission, where acquisition of immunity to malaria is very important, it has been argued that vector control (without vector eradication) could, in the long run, make the situation worse by preventing the normal build-up of immunity. However, our data from Tanzania do not support this idea--3-4 years after provision of nets (which are re-treated annually) young children are still showing clear health benefits; older children are not "paying" for this by showing worse impact of malaria. There is less malaria morbidity in a highland area where malaria transmission is about 15x less intense than in a nearby lowland area. The per cent impact of treated nets malaria morbidity in both area was very similar. At present only pyrethroids are used for net treatment which suggested that emergence of pyrethroid resistance would have a disastrous effect. However, in West Africa, where there is now a high frequency of the kdr resistance gene in Anopheles gambiae, it is reported that treated nets continue to have a powerful impact on vector populations. In Tanzania, pyrethroid resistance has not been detected in malaria vectors, but it has emerged in bedbugs after seven years use of treated nets. PMID- 15119066 TI - Malaria genome project and its impact on the disease. AB - Malaria remains uncontrolled to-date due to lack of effective parasite and vector control strategies. With the completion of the host, parasite and vector genome projects more suitable and effective disease control measures can be achieved. Here we have reviewed the Plasmodium falciparum genome project and its impact on malaria research in future. The parasite genome project has revealed certain metabolic pathways which can be targeted to develop antimalarial drugs. It has also identified large number of potential antigens for the future potential vaccines. Now the researchers in the malaria field can plan to take up the studies, which can yield more fruitful results within the limited financial resources using bioinformatics, proteomics, structural, functional and comparative genomics, etc. PMID- 15119067 TI - Malaria morbidity and mortality in Uganda. PMID- 15119068 TI - Biolarvicides in vector control: challenges and prospects. AB - Biolarvicides, based on mosquitocidal toxins of certain strains of Bacillus sphaericus and Bacillus thuringiensis var israelensis H-14 (Bti) are highly effective against mosquito larvae at very low doses and safe to other non-target organisms. During past two decades various biolarvicide formulations produced in India and abroad have been tested at Malaria Research Centre and some formulations have undergone large-scale operational trials. Biolarvicide formulations of B. sphaericus are useful in the control of Culex and certain Anopheles spp, such as An. stephensi and An. subpictus, but not much effective against An. culicifacies and almost ineffective against Aedes aegypti. Repeated application of B. sphaericus in the same habitat, however, results in the development of resistance in larvae of target mosquitoes. In view of its low specificity for An. culicifacies and the potential for resistance in An. stephensi, B. sphaericus has limited prospects for control of malaria vectors. However, with some resistance management, B. sphaericus can still be used against Culex mosquitoes. On the other hand Bti formulations, which have broader spectrum of activity against Aedes, Culex and Anopheles spp, have not shown significant development of resistance in mosquitoes but their activity in field, particularly against surface feeding anopheline larvae is affected by various bioenvironmental factors, thus requiring weekly application in most habitats. To overcome this problem development of slow release formulations and genetically engineered biolarvicides by transplanting mosquitocidal toxin genes of Bti and B. sphaericus in some other environmentally compatible organisms have been investigated by different scientists. PMID- 15119069 TI - Bio-efficacy and operational feasibility of alphacypermethrin (Fendona) impregnated mosquito nets to control rural malaria in northern India. AB - Bio-efficacy and operational feasibility of alphacypermethrin treated nets was evaluated in certain villages of District Ghaziabad (U.P.). Results revealed that poly-filament nylon nets treated with alphacypermethrin suspension concentrate (g/l) formulation @ 25 mg/m2 has shown repellent action (26.5 +/- 8.1), excito repellent action (93.7 +/- 8.1) and killing action (100%) against An. culicifacies landed on treated nets. Significant reduction in indoor resting density of An. culicifacies in human dwellings was also observed in treated nets village (p < 0.05). Instant killing action of treated nets did provide complete protection to inhabitants sleeping inside the net from An. culicifacies bites. The persistent use of nets by the inhabitants has also resulted significant reduction in malaria cases (p < 0.05). It was interesting to note that not even a single case of falciparum malaria was observed after distribution of treated nets in spite of the fact that at no point of time cent per cent compliance of net usage was observed during the study period. Bio-assay tests revealed that treated nets can produce up to 70% mortality in An. culicifacies for about 22 weeks and as such only one treatment with insecticide is required in a year in seasonal transmission area to protect from malaria. The study also revealed that treated nets can be stored at room temperature for about 10 months without loosing their efficacy suggesting thereby that malaria outbreaks can be tackled by the nets if adequate treated nets are stored in core problem districts. It was also revealed that An. culicifacies is a late night biter and as such treated nets can be used successfully against this species. PMID- 15119070 TI - Bioecology of An. philippinensis in Andaman group of islands. AB - Studies on bioecology of An. philippinensis a vector of malaria was carried out in eight islands of the Andaman group. It was found that An. philippinensis preferred to rest and bite outdoors. Maximum biting was observed during 1800 to 2100 hrs on both cattle and human bait. The maximum breeding of An. philippinensis was recorded in slow moving stream followed by ponds with vegetation. The breeding sites infested with Chara and Spirogyra algae were most favourable whereas, with Lemna and Oscillatoria were unfavourable for the proliferation of this species. The breeding association of An. philippinensis was found with other seven anopheline species in different breeding habitats. PMID- 15119071 TI - Evaluation of botanicals as repellents against mosquitoes. AB - Repellent properties of three plant extracts--essential oil (steam distillate) of Zanthoxylum limonella (fruits), Citrus aurantifolia (leaf) and petroleum ether extract of Z. limonella (fruits) were evaluated as repellent against Aedes (S.) albopictus mosquitoes in mustard (Dhara) and coconut (Parachute) oil base under laboratory conditions. Three concentrations--10, 20 and 30% of the repellents were evaluated. Repellents in mustard oil afforded longer protection time against the bites of Aedes (S.) albopictus mosquitoes than those in coconut oil. At 30% concentration, 296-304 min protection time was achieved by the test repellents in mustard oil base while repellents in coconut oil exhibited 223.5-245 min protection time at the same concentration. Oil of Z. limonella gave the highest protection time against the bites of Aedes (S.) albopictus mosquitoes at all the concentrations than other herbal repellents tested both in mustard and coconut oil. PMID- 15119072 TI - Irritant effect, prevention of blood feeding and toxicity of nets impregnated with different pyrethroids on An. stephensi. AB - A resistant strain of An. stephensi was confirmed to have high resistance to knock down and kill by four different pyrethroids in bioassays in comparison with a susceptible strain. Permethrin, lambda-cyhalothrin and deltamethrin were more irritant to the susceptible strain than the resistant as judged by time for first take-off, but nets treated with alphacypermethrin appeared to be more irritant to the resistant An. stephensi than the susceptible. With all the pyrethroids tested, the resistant strain spent a longer time in contact with a treated net, which was in contact with a human arm, than did the susceptible strain. With permethrin the resistant strain fed significantly more successfully through the treated netting than did the susceptible strain. With deltamethrin there was a non-significant tendency in the same direction in comparing the two strains. However, with alphacypermethrin there was a non-significant tendency in the reverse direction. After 15 min in the cage which tested for the ability to feed through a pyrethroid treated net, observed mortality was higher with the susceptible than the resistant strain. Thus there was no sign that the longer resting of the resistant strain on treated netting would compensate for the fact that a higher dose was needed to kill this strain. Such compensation has been suggested with West African An. gambiae where treated nets continue to work well against a highly resistant wild population. However, this does not seem to apply to our resistant An. stephensi. PMID- 15119073 TI - Genetics of resistance to permethrin in Anopheles stephensi. PMID- 15119074 TI - Drug resistant falciparum malaria and the use of artesunate-based combinations: focus on clinical trials sponsored by TDR. AB - Antimalarial drug resistance has now become a serious global challenge and is the principal reason for the decline in antimalarial drug efficacy. Malaria endemic countries need inexpensive and efficacious drugs. Preserving the life spans of antimalarial drugs is a key part of the strategy for rolling back malaria. Artemisinin-based combinations offer a new and potentially highly effective way to counter drug resistance. Clinical trials conducted in African children have attested to the good tolerability of oral artesunate when combined with standard antimalarial drugs. The cure rates of the different combinations were generally dependent on the degree of resistance to the companion drug. They were high for amodiaquine-artesunate, variable for sulfadoxine/pyrimethamine-artesunate, and poor for chloroquine-artesunate. PMID- 15119075 TI - Interrupting malaria transmission by genetic manipulation of anopheline mosquitoes. AB - Malaria ranks among the deadliest infectious diseases that kills more than one million persons every year. The mosquito is an obligatory vector for malaria transmission. In the mosquito, Plasmodium undergoes a complex series of developmental events that includes transformation into several distinct morphological forms and the crossing of two different epithelia--midgut and salivary gland. Circumstantial evidence suggests that crossing of the epithelia requires specific interactions between Plasmodium and epithelial surface molecules. By use of a phage display library we have identified a small peptide SM1--that binds to the surfaces of the mosquito midgut and salivary glands. Transgenic Anopheles stephensi mosquitoes expressing a SM1 tetramer from a blood inducible and gut-specific promoter are substantially impaired in their ability to sustain parasite development and transmission. A second effector gene, phospholipase A2, also impairs parasite transmission in transgenic mosquitoes. These findings have important implications for the development of new strategies for malaria control. PMID- 15119076 TI - Markers for population genetic analysis of human plasmodia species, P. falciparum and P. vivax. AB - Present report deals with the genetic diversity existing among the field isolates of Plasmodium falciparum and P. vivax in India. Isoenzymes and molecular markers were used to analyse field isolates of P. falciparum and P. vivax. High level of length polymorphism was observed in repeat nucleotide sequences of MSP-1, MSP-2 and GLURP in P. falciparum isolates and CSP, GAM-1 and MSP-3 alpha in P. vivax isolates. In study populations a high proportion of isolates (up to 60%) were comprised of more than one genetically distinct parasite type--multiclonal. Presence of identical allelic forms of enzyme and DNA variations in different geographical areas and in different years suggest that isolates belong to a single random mating population of P. vivax and P. falciparum. Observed random combination of alleles in the field isolates suggest the unlinked nature of loci studied. Study supports the feasibility of using molecular markers for the identification of recrudescence in P. falciparum from fresh infection. PMID- 15119077 TI - How specific is the immune response to malaria in adults living in endemic areas? AB - It is documented that people living in malaria endemic areas acquire immunity against malaria after repeated infections. Studies involving passive transfer of IgG from immune adults to the nonimmune subjects have shown that circulating antibodies play an important role, and that immune adults possess protective antibodies, which susceptible malaria patients do not. Through a differential immunoscreen, we have identified several novel cDNA clones, which react exclusively and yet extensively with immune sera samples. Specific antisera raised against the immunoclones inhibit the growth of parasites in culture. The clones studied so far turn out to be novel conserved Plasmodium genes. In order to study the response of sera of adults from malaria endemic areas of India and Africa to these immunogens, we carried out ELISA assays using these immunopeptides, other P. falciparum specific antigens, peptides, antigens from other infections such as mycobacterial infections and other proteins such as BSA. Children from the same areas and normal healthy urban people showed very little activity to each of these categories. A large percentage of adults from endemic areas responded positively to all the malarial immunogens tested. However, the same persons also showed high response to other antigens and proteins as well. The implications of these results are reported in this paper. PMID- 15119078 TI - Assessment of therapeutic efficacy of chloroquine and sulphadoxine-pyrimethamine in uncomplicated falciparum malaria. AB - A standardised protocol has been developed by World Health Organization (CDS/RBM/2002) to assess the efficacy of common antimalarials in the treatment of clinically manifested infection with uncomplicated P. falciparum malaria for areas with low to moderate transmission. The therapeutic efficacy protocol is based on clinical and parasitological responses of the patients and it has the purpose of determining the practical efficacy of the drug regimen in study areas with the ultimate objective of ascertaining its continued usefulness or the necessity for replacing it in the routine treatment. Present study has been conducted at seven sites--Kathiatali and Simonabasti of District Nowgaon, Assam; Sonapur and Boko of District Kamrup, Assam; Keonjhar Town, Padampur and Basudebpur of District Keonjhar, Orissa. In order to reduce the patient recruitment time, health centre close to well-defined community was identified to conduct the activities at peak malaria season by selecting local pockets and organising mobile clinics. Microscopically confirmed cases of P. falciparum were enrolled according to the criteria for inclusion and exclusion. Treatment with recommended drug was given under supervision and a follow-up schedule at various intervals for 28 days was maintained. In chloroquine (CQ) study areas, wherever patients showed treatment failure, they were treated with second line drug- sulphadoxine-pyrimethamine (SP) combination and then followed-up as per study protocol. It was observed that 30% cases showed treatment failure to CQ in District Nowgaon, where revised drug policy has already been introduced. In Kamrup district, treatment failure with CQ was found to be less than 25%, which denotes the said regimen is still effective. Almost all the patients from Padampur and Basudebpur of District Keonjhar responded to CQ, treatment failure was noticed only in two patients (3%). The antifolate combination found to be fully effective as second line and also as first line wherever revised drug policy has been introduced. PMID- 15119079 TI - Evaluation of repellent action of Cymbopogan martinii martinii Stapf var sofia oil against Anopheles sundaicus in tribal villages of Car Nicobar Island, Andaman & Nicobar Islands, India. AB - A field study was carried out to evaluate the mosquito repellent action of Cymbopogan martinii martinii Stapf var sofia oil in tribal village of Car Nicobar Island. Results revealed that application of 1 ml of the oil provided 98.7% protection in indoor and 96.52% in outdoor conditions during 12 h period of observation from the bites of An. sundaicus. Cymbopogan martinii martinii Stapf var sofia oil is an indigenous product, its application is safe and can be widely used for protection from malaria. PMID- 15119080 TI - Testing of newly developed glycophospholipid antigen for the detection of P. falciparum malaria by laser light immunoassay in endemic and non-endemic areas. AB - A glycophospholipid (GPL) antigen isolated from Plasmodium falciparum culture supernatant has been tested for its antigenicity. Detection of malaria positive known blood samples and unknown field samples from endemic and non-endemic areas were compared. In this study laser light scattering immunoassay (LIA) was used for the detection of P. falciparum malaria. Test results of control (malaria negative samples from Surat) were compared with known positive samples and unknown malaria positive field samples. A positive correlation has been observed (97%) in falciparum positive samples from laboratory and unknown samples from endemic area (Haldwani) by LIA method using GPL antigen. From the results of the study it was found that GPL antigen has a better antigenic property and can detect almost all the cases of Pf malaria by LIA method. PMID- 15119081 TI - Hospital based study of malaria in Ratnagiri district, Maharashtra. PMID- 15119082 TI - An inventory of Culicidae diversity in Haryana state. PMID- 15119083 TI - Spotlight on success. PMID- 15119084 TI - Levelling the playing field. PMID- 15119085 TI - Agenda for change: implications for the voluntary sector. PMID- 15119086 TI - Consent to treatment when capacity is absent. Part 2: Need for change. PMID- 15119087 TI - A guide to peer appraisal. AB - The experience of peer appraisal can be a positive one. The process can help to establish a shared understanding of what is needed for individual, team and organisational objectives to be achieved. Traditional appraisals tend to be about 'me'; the peer approach is about 'us'. The benefits can include a greater sense of ownership, empowerment and responsibility, more support within peer groups and better communication. It may be considered for nurses in all fields. PMID- 15119088 TI - Managing diversity. PMID- 15119089 TI - Reference queries. PMID- 15119090 TI - Take it to the top. PMID- 15119091 TI - What's in a name? PMID- 15119092 TI - A pilot study of patients' views of an oral health scoring system. AB - An oral health scoring system (Oral Health Score: OHS) has been designed to provide a numerical measure of the overall state of a patients oral health by means of a series of simple clinical examinations. OBJECTIVE: To assess, by means of a questionnaire, patients opinions of the value of the OHS. METHODS: Eight general dental practitioners who used the OHS were asked to participate in the project. The participating dentists were requested to explain the aims of the project to 50 patients. Patients who agreed to participate were asked to complete a questionnaire after a visit during which the OHS was used. These patients were provided with a reply-paid envelope for the return of the questionnaire. RESULTS: Completed usable questionnaires were received from 315 patients. Of respondents, 97% considered that the OHS gave them a better understanding of the condition of their mouth and 98% considered that the OHS was a good method for communication between dentist and patient. CONCLUSION: The results of the present study indicate that, within the study group, the OHS is considered by a large majority of patients to be a good method for communication of the patient's oral health between dentist and patient. PMID- 15119093 TI - Understanding the issue of 'informed consent' in dental treatment. AB - The concept of consent to treatment is increasingly becoming contested in United Kingdom courts of law. Any practitioners who cannot demonstrate that a patient has properly consented to treatment are laying themselves open to litigation. This paper demonstrates that valid consent is not as straightforward as may be assumed and that a patient can easily challenge a standardised approach to obtaining consent. Current Department of Health guidelines on obtaining consent are discussed, and changes in the USA. Europe and Australia are brought into focus with regard to the situation in the UK. PMID- 15119094 TI - Internet-derived patient information on common oral pathologies: is it readable? AB - BACKGROUND: The evolution of the Internet has made available to the public vast reserves of information, and it represents a tremendous educational resource. However, it is obviously important that information directed at patients is readable and understandable. OBJECTIVE: This study aimed to evaluate the readability of patient-related Internet sites presenting information on dental caries, periodontal disease, oral cancer and mouth ulcers. METHODOLOGY: Four separate Internet searches seeking patient information on these pathologies were conducted in May 2003. In total, 5127 sites were identified. Within each search, the first 25 sites identified as of relevance to patients seeking information were downloaded (100 in total), and the readability determined using the Flesch reading score and the Flesch-Kinkaid reading level. RESULTS: A wide range of reading ease scores was obtained, equating to reading ages from seven years to university entry level. The mean reading age of the sites evaluated was 10-11 years of age, higher than the estimated reading age of the UK population in general (nine years). Scores were broadly similar across different pathologies. CONCLUSIONS: Practitioners and other members of the dental team involved in patient education should be aware that additional advice and support may be required to help patients interpret material downloaded from the Internet. Furthermore, those practices presenting educational material on their practice website should bear in mind the need to ensure that it is pitched at a level appropriate to the intended audience. PMID- 15119095 TI - An investigation of aspects of design of partial dentures. AB - PURPOSE: The purpose of the study was to establish the incidence of the prescription of partial denture design by dentists and also to establish the nature of clasp design, if prescribed. BASIC PROCEDURES: Data relating to cobalt chromium partial dentures were collected from filed slips from a dental laboratory that provided fabricated cobalt-chromium partial dentures for hospital and general dental-practitioners. The data collected related to whether or not there was a design for the prosthesis and, also, to the design of clasp (gingivally or occlusally approaching). MAIN FINDINGS: Only 39% of slips had evidence of a partial denture design and of those, 41.5% involved the prescription of an inappropriate design of clasp. PRINCIPAL CONCLUSIONS: The incidence of partial denture design by a group of dentists using a commercial dental laboratory is unacceptably low. Despite evidence advocating the use of gingivally-approaching clasps on teeth anterior to (and including) premolars, there is still an unacceptably high level of the use of occlusally-approaching designs. PMID- 15119096 TI - Clinical governance. What is it and how can it be delivered in dental practices? AB - This paper describes how a Scottish general dental practitioner carried out a literature search to inform his teaching on the topic of clinical governance in general dental practice. He found relatively few publications on the topic, most of which were descriptive rather than reports of scientific studies. Nevertheless, he has used the results of the literature search to inform his teaching. His opinion on what clinical governance is and how it can be introduced follows a brief description of the literature search. PMID- 15119097 TI - Take courage. PMID- 15119098 TI - Depression: are you treating to remission? PMID- 15119099 TI - Foreign body induced appendicitis. PMID- 15119100 TI - Respiratory failure secondary to methemoglobinemia induced by benzocaine: a case report. PMID- 15119101 TI - Breast cancer--the disease many women fear the most. PMID- 15119102 TI - 2004 shaping up to be another busy year for civil justice reform. PMID- 15119103 TI - The challenges of medical genetics and the primary care practitioner. PMID- 15119104 TI - Can they really take away my hospital privileges? PMID- 15119105 TI - Partisan politics do not belong in Missouri Medicine. PMID- 15119106 TI - Enzyme replacement therapy for lysosomal storage disorders: successful transition from concept to clinical practice. AB - Lysosomal disorders offer a striking example of how basic scientific research has been translated into effective medical intervention. The successful application of enzyme replacement therapy for several lysosomal disorders in animal models and in human disease offers hope for everexpanding abilities to counter the effects of these disorders. PMID- 15119107 TI - Genetic susceptibility to breast and ovarian cancer: risk assessment, genetic counseling, and management issues. AB - Genetic testing for disease susceptibility genes is available for a large number of genetic disorders. With the expected shortage of trained genetic professionals to offer risk assessment, counseling, testing, and management to patients, primary care physicians will be asked to provide these services. In this review, we discuss the genetic basis of breast and ovarian cancer susceptibility and outline specific recommendations for clinical practice. PMID- 15119108 TI - Neurocutaneous syndromes. AB - Neurocutaneous syndromes are progressive multisystem disorders in which the dermatological findings may be apparent before the neurological signs and symptoms appear. Timely diagnosis is important for early detection of serious complications such as malignancies, for prompt interventions for learning or developmental problems, and for genetic counseling. PMID- 15119109 TI - The ethical, legal and psychosocial challenges of genetic testing: implications for primary medical care. AB - Recent advances in genetic information and technology have led to an explosion in molecular testing for mutations that cause human disease. Test results are likely to improve medical management and/or help individuals make important life and reproductive decisions. Unlike other laboratory testing, unique complexities and challenging psychosocial, legal and financial ramifications often accompany genetic tests. Some of the challenges associated with molecular testing and examples illustrating potential ethical, psychosocial and legal complexities are presented here. PMID- 15119110 TI - Prenatal screening for birth defects: an update. AB - This article summarizes current second trimester prenatal screening methods, and describes recently developed first trimester screening tools. The advantages and potential pitfalls of first trimester screening are outlined. PMID- 15119111 TI - Resources of genetic education for non-geneticists. AB - A variety of educational efforts intended for primary care physicians have been developed or are at the stage of planning, from workshops, conferences, CD-ROMs, and educational websites to formal research training. This article summarizes these resources. PMID- 15119112 TI - Ordering errors by first-year residents: evidence of learning from mistakes. AB - OBJECTIVE AND DESIGN: To study patterns of ordering errors, we reviewed charts of patients cared for by first-year residents at our community-based teaching hospital. RESULTS: In month one, 39 of 1248 orders (3.1%) were rated as errors. In contrast, 9 of 1072 orders (0.84%) in month six were rated as errors (p < 0.001). Of the 48 errors identified, 36 (75%) related to medications (choice, dose, route, frequency, or duration). Over half (53%) of these errors involved antimicrobials, pulmonary medications, or diabetes-related drugs. For all errors, half (24/48) were identified by attending physicians, with hospital support staff noting most of the remaining errors (43.8%). Errors were ultimately corrected by the first-year resident in over half of the cases (52%); attending physicians corrected 16 of 39 errors (41%) in month one but only one of nine errors (11%) in month six. Errors by first-year residents were primarily attributed to inattention (45.8%) or deficits in clinical knowledge (43.8%). No adverse effects resulted from any of the ordering errors identified. Primary consequences of errors included inconvenience to staff (50%), delay in treatment or diagnosis (31.3%), or receiving unneeded medication (18.8%). CONCLUSIONS: The number of medical errors decreases with experience. Our results highlight the importance of clinical supervision during the initial months of training. PMID- 15119113 TI - Detection of pathogenic Ehrlichia in ticks collected at acquisition sites of human ehrlichiosis in Missouri. AB - We collected 385 ticks from sites in Missouri associated with human monocytic ehrlichiosis. Using PCR, we detected E. chaffeensis or E. ewingii in 2 of 19 pools of adult Amblyomma americanum, 0 of 32 pools of Dermacentor variabilis, and 6 (18%) of 39 pools of unspeciated nymphal ticks from 3 of 6 sites associated with disease and one site not associated with disease. We also detected a variant of A. phagocytophila in one nymph pool. PMID- 15119114 TI - [Chemoprevention of colorectal cancer]. AB - Although colorectal cancer is one of the most preventable forms of cancer, it remains the second leading cause of cancer death worldwide. Primary prevention involves the identification and elimination of factors, which cause or promote colorectal cancer. The goal of screening is to prevent colorectal cancer mortality through the detection and treatment of premalignant adenomas and curable-stage cancer. Most colorectal cancers are believed to arise from adenomatous polyps. Early identification and removal of adenomas can prevent the development of colorectal cancer. Chemoprevention is the use of specific chemical compounds to prevent, inhibit, or reverse carcinogenesis. Several chemoprevention options have been investigated and confirmed as effective. Aspirin and other nonsteroidal anti-inflammatory drugs are the most widely studied agents, their use has been consistently associated with reduction in the risk of mortality and the incidence of colorectal adenomas and cancers. The selective cyclooxygenase-2 (COX-2) inhibitors (coxibs) have been demonstrated to decrease the number and the size of polyps in patients with familial adenomatous polyposis syndrome. Because the gastrointestinal toxicity of coxibs is lower, it might be safer than aspirin or other non-selective nonsteroidal anti-inflammatory drugs for long-term use. This review aims to summarize the recent theoretical and practical advances in the chemoprevention of colorectal cancer. PMID- 15119115 TI - [Role of lipid peroxidation in non-alcoholic steatohepatitis]. AB - The liver steatosis is a frequent human disease. The most frequent cause of the process is the alcohol consumption. But it also may arise without significant alcohol abuse. This pathogenetic process is called non-alcoholic steatohepatitis (NASH), that is characterized by the same conditions like the alcoholic steatohepatitis. In the pathogenesis of the NASH various factors participate i.e. obesity, diabetes mellitus type II, hyperlipidaemia, pregnancy, variable chemotoxins, parenteral nutrition, jejunoileal bypass, chronic inflammatory diseases, protein deficient nutrition and inborn metabolic diseases. Pathobiochemically the process consists of oxidative stress and lipid peroxidation. This condition comes from the progressive accumulation of the free fatty acids in mitochondria and from the induction of cytochrome P450, CYP 2E1 isoform in hepatocytes and Kupffer cells. The free fatty acids and ketons can cause the induction of CYP 2E1 system, that is why diabetes mellitus and obesity are the two most important factors in the NASH pathogenesis. This article is concerned mainly in the explanation of NASH pathomechanism. PMID- 15119116 TI - [Treatment of acute ST-elevation myocardial infarction with primary percutaneous coronary intervention]. AB - INTRODUCTION: Treatment of acute myocardial infarction has changed significantly in recent years. Since January 1st, 2003 primary coronary intervention has been organized in Budapest. The authors show the results during the implementation of this method in the first 6 months with emphasizing its importance. PATIENTS: In the center which was developed specially for catheter interventions in myocardial infarction, 282 patients were admitted during this period with the suspicion of acute ST-elevation myocardial infarction. 266 (94.3%) of them were proved to have an infarction. Of these, 260 had a lesion amenable for percutaneous intervention. RESULTS: Success rate of catheter-based revascularization was 96.5%. 4 patients underwent open-heart surgery. Adverse events were: severe bleeding 1.1%, in hospital mortality 4.5%. CONCLUSION: In our center the major outcomes are similar to the results of large international centers. PMID- 15119117 TI - [Assessment of patient satisfaction for the improvement of emergency medical services]. AB - Quality expectations of emergency medicine are not fully formed yet. However, client satisfaction examinations proved to be a valid tool for assessment of different parts of medical care. To define process parameters turning to efficiently expand of maintenance-quality the authors developed and applied a client satisfaction questionnaire in Szent Imre Hospital Emergency Department. Answers for client satisfaction questionnaire collation of characteristics of patients' demographic data, sociological situation, and quality of life were studied by multinomial regression analysis in a prospective pilot study. The returned questionnaires were represented in 28% of the examined patient group. To achieve the greatest improvement in patients satisfaction, painkilling, patients information, and inter-relationship between patients and providers would be corrected in the emergency department. Compound client satisfaction questionnaire was proved to be a susceptible tool for identified the relevant pars of the process quality. PMID- 15119118 TI - [Treatment of non-alcoholic steatohepatitis]. PMID- 15119119 TI - [Treatment of diabetic angiopathies]. PMID- 15119120 TI - The road to integrated care: commitment is the key. Tennessee CMHC demonstrates promise of co-located behavioral and primary care. PMID- 15119121 TI - Small project reveals large cultural barriers to integrated care. PMID- 15119122 TI - Energizing young professionals through effective training. NAATP award winner demonstrates a culture of quality. PMID- 15119123 TI - Clinicians say they're twisting in wind during FDA review of SSRIs. PMID- 15119124 TI - When doing the right thing might be wrong. Research questions value of a widely used crisis response. PMID- 15119125 TI - A mission that stresses the importance of home. Interview by Linda Watts Jackim. PMID- 15119126 TI - Are minimum staff ratios needed? PMID- 15119127 TI - Was reclassifying cannabis a wise decision? PMID- 15119128 TI - Patient choice. PMID- 15119129 TI - 'We are people who can get things done'. PMID- 15119130 TI - The importance of improving awareness of colorectal cancer. AB - The function of the bowel and the incidence of bowel cancer are issues that we do not want to think about. Embarrassment around the symptoms, ignorance, and fear, are barriers that need to be overcome in order to reduce the number of people affected by the disease. Colon Cancer Concern's awareness campaign aims to start members of the public talking about this killer disease and acting upon their concerns. PMID- 15119131 TI - What you need to know about ... typhoid. PMID- 15119132 TI - The link between mental health problems and violent behaviour. AB - Fear of violence in relation to people with mental illness is common and affects the way society views and treats such individuals. This article reviews literature on mental illness and violence with the aim of clarifying whether there is a link between the two. While higher rates of violence have been found among people with mental illness, most studies have flaws and their results should be viewed with caution. Further research is required particularly in connection with people with serious mental illness. We also need to develop preventative strategies by providing appropriate support for people with mental health problems. PMID- 15119133 TI - Development of a district-wide teledermatology service. AB - Demand for dermatology services is increasing, resulting in changes in service provision and the role of nurses. Technological advances have led to the development of telemedicine. This article describes how four primary care trusts and the dermatology department at Queen's Medical Centre developed a district wide teledermatology service. The service was led by nurses and a GP with a special interest in dermatology. PMID- 15119134 TI - Why good communication skills are important for theatre nurses. AB - Excellent communication skills, both verbal and nonverbal, are needed throughout the perioperative experience to educate patients undergoing surgery and so ease their anxiety. In addition, good communication with theatre colleagues, the multidisciplinary team, and other departments is vital to achieve technical excellence and individualised, holistic patient care. This article discusses the importance of effective communication and describes how it can be improved in the perioperative setting. PMID- 15119135 TI - The use of maggots as a new treatment in the community. AB - Since maggots were reintroduced into modern wound management practice, published papers have described their mode of action in the treatment of wounds such as pressure sores and leg ulcers (Thomas et al, 1996a; Thomas et al, 1996b), diabetic ulcers (Jones and Thomas, 2000; Johnson, 1999; Murray and Benbow, 1999; Rayman et al, 1998; Evans, 1997; Mumcuoglu et al, 1997; Thomas et al, 1996a), traumatic injuries (Thomas et al, 1996b), burns (Namias et al, 2000) and surgical wounds (Fear et al, 2003; Jones and Champion, 1998; Young, 1997). PMID- 15119136 TI - Guidance on pressure ulcer risk assessment and prevention. AB - Pressure ulcers remain a problem throughout Europe, with prevalence figures ranging from 8.3 per cent to 22.9 per cent (Clark and deFlour, 2002). In 2001 the National Institute for Clinical Excellence issued a clinical guideline on pressure ulcer risk assessment and prevention (NICE, 2001). However, this guidelines has recently been reissued with additional information on pressure relieving devices (NICE, 2003a; NICE, 2003b). PMID- 15119137 TI - The use of a pressure ulcer risk assessment tool for children. AB - The need for pressure ulcer risk assessment has been acknowledged in adult nursing for many years. However, in paediatrics there is a misconception that children do not develop pressure ulcers (Mills, 1998). This misconception has been challenged in recent years with a growing awareness that children have specific risk factors that predispose their skin to pressure damage (Willock et al, 2000; Mills, 1998; Waterlow, 1998, 1997; Jones, 1997; Pickersgill, 1997; Zollo et al, 1996; Bedi, 1993). PMID- 15119138 TI - Delayed wound healing in patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is an inflammatory disease that causes pain, swelling, stiffness, and loss of function in the joints. It occurs in a symmetrical pattern, which means that if one knee or hand is involved, the other is also (Box 1). It is a chronic, progressive, inflammatory tissue disorder of unknown origin and in addition to joint stiffness, ankylosis (fixation of a joint), and associated joint deformity, patients may have systemic involvement of the eyes, kidneys, chest, and lungs (Ryan, 1995). It is thought that the autoimmune component of the disease can have significant multi-system effects, including scleritis (inflammation of the white of the eye, the sclera), pericarditis, pleural effusions, vasculitis (patchy inflammation of the walls of small blood vessels), and skin ulceration (Oliver and Mooney, 2002). PMID- 15119139 TI - How to ... network. PMID- 15119140 TI - Moving forces. PMID- 15119141 TI - Welcome to planet nursing. PMID- 15119142 TI - The patient factor. PMID- 15119143 TI - On the up. PMID- 15119144 TI - Schools of thought. PMID- 15119145 TI - Nurse practitioner education: learning from students. AB - AIM: To describe the experiences of nurse practitioner (NP) students at Bournemouth University. METHOD: Grounded theory methodology guided the data collection and analysis. A theoretical model was constructed that represented 'role transition' for NP students. RESULTS: The social environment of the participants affected role transition and learning and two major categories were evident: experience in the workplace and experience in the academic environment. A third category, 'pioneering spirit', described the participants' personal attributes that influenced their transformation. CONCLUSION: The results have prompted a review of the current curriculum, mode of delivery and teaching resources. The findings have implications for current and future courses and demonstrate the value of eliciting the students' view. PMID- 15119146 TI - Down's syndrome: a biopsychosocial perspective. AB - This article examines the biological, psychological and social effects of Down's syndrome and disability on children who have this condition, and on their families or carers. PMID- 15119147 TI - Managing risk in the perioperative environment. AB - This article provides an overview of managing risk in surgical patients in the perioperative environment. PMID- 15119148 TI - Take it to the top. PMID- 15119149 TI - The tsar's progress. Interviews by Hazel Heath. PMID- 15119150 TI - Seeking refuge. PMID- 15119151 TI - HelpAge International rebuilding older people's lives. PMID- 15119152 TI - Prevention of falls in older people: the Weymouth and Portland project. PMID- 15119153 TI - Touch: a fundamental aspect of communication with older people experiencing dementia. PMID- 15119154 TI - Parkinson's disease, palliative care and older people: Part 2. AB - In the second of two articles on palliative care for people with Parkinson's disease, the authors consider other aspects of care relevant to this complex stage. They conclude that better understanding of its complexity, its innate variability and the roles extended team members can play help to improve the care given to patients. PMID- 15119155 TI - ACE inhibitors. PMID- 15119156 TI - End of an era. PMID- 15119157 TI - Mapping the journey. PMID- 15119158 TI - Clinical observation in 102 cases of chronic pelvic inflammation treated with qi jie granules. AB - OBJECTIVE: To observe the therapeutic effects of Qi Jie Granule [see: symbol text] on chronic pelvic inflammation. METHOD: The therapeutic effect, T lymphocytic subgroups and indexes of blood rheology were observed when 102 cases of chronic pelvic inflammation in the treatment group were treated with Qi Jie Granule, and another 70 cases were treated with Qian Jin Pian as the controls. RESULTS: The total effective rate was 96.08% in the treatment group, but 84.29% in the control group with a significant difference between the two groups (P < 0.01). Qi Jie Granule was also found effective in improving blood viscosity and regulating T-lymphocytic subgroups, and the difference before and after the treatment was also very significant (P < 0.05 or P < 0.01). CONCLUSION: The nature of chronic pelvic inflammation is qi deficiency and blood stasis in accordance with the theory of traditional Chinese medicine (TCM). As a drug that is indicated for chronic pelvic inflammation, its mechanism may be related to the improved blood circulation, accelerated inflammatory absorption and regulated immune function. PMID- 15119159 TI - Dr. Zhu Hongming's experience in treating chronic gastritis. PMID- 15119160 TI - Treatment of chronic bronchitis with modified ma xing shi gan tang and er chen tang. PMID- 15119161 TI - Treatment of post-menopausal urethral syndrome with the formula niao chang shu--a report of 40 cases. PMID- 15119162 TI - The effects of acupuncture in treatment of coronary heart diseases. AB - A clinical study on acupuncture at point Neiguan (PC 6) was conducted on the basis of successful treatment of patients with angina pectoris and acute myocardial infarct. The effective rate (91.3%) was found to be far superior to isosorbide dinitrate and nifedipine (P < 0.01). The experimental studies in animal models demonstrated that electro-acupuncture could decrease the elevated ST segments in ECGs and reduce the infarct areas induced by coronary ligation. PMID- 15119163 TI - Treatment of mental depression due to liver-qi stagnancy with herbal decoction and by magnetic therapy at the acupoints--a report of 45 cases. PMID- 15119164 TI - Treatment of melancholia in Germany by acupuncture method for resuscitation. PMID- 15119165 TI - Clinical application of moxibustion. PMID- 15119166 TI - Treatment of pseudobulbar paralysis by scalp acupuncture and sublingual needling. PMID- 15119167 TI - Effects of electroacupuncture plus intra-carotid drug injection on rheoencephalogram in patients with cerebral infarction. AB - PURPOSE: To investigate the mechanism of electroacupuncture (EA) plus intra carotid drug injection for treating cerebral infarction. METHODS: Rheoencephalogram was recorded with a RG-2B type of bridge rheoencephalograph and findings were compared before and after the treatment. RESULTS: After the treatment, the prolonged rising time was shortened, and the decreased amplitude obviously elevated. CONCLUSION: The therapy can dilate cerebral blood vessels, increase the cerebral blood flow, and improve the elasticity of cerebral blood vessels, leading to sufficient blood and oxygen supply in the ischemic brain tissues and to restoration of their functions. PMID- 15119168 TI - Introduction to the points singly used for stiff neck. PMID- 15119169 TI - Comparison of the effects of electroacupuncture plus cupping with that of the electrical pulse therapy for different types of cervical spondylopathy. AB - OBJECTIVES: To observe and compare the therapeutic effects of electroacupuncture (EA) plus cupping and the electrical pulse therapy on different types of cervical spondylopathy. METHODS: 182 cases of cervical spondylopathy confirmed by X-ray or CT examination were divided into 4 types, and treated with EA plus cupping in Group A of 90 cases and with moderate or low electrical pulses in Group B of 92 cases. The therapeutic effects were compared after 3 courses of treatments. RESULTS: For the vertebroarterial type and the nerve root type of cervical spondylopathy, the effect in Group A was better than that in Group B, while for the sympathetic type of cervical spondylopathy, the effect in Group B was superior to that in Group A. CONCLUSION: Appropriate therapies should be adopted for different types of cervical spondylopathy. PMID- 15119170 TI - Sixty-two cases of simple obesity treated by acupuncture combined with massage. PMID- 15119171 TI - Frequent ventricular extrasystole treated by needling neiguan (PC 6) plus oral administration of mexiletine--a report of 30 cases. AB - PURPOSE: To observe the therapeutic effect of needling at Neiguan (PC 6) plus oral administration of mexiletine. METHODS: 60 cases of frequent ventricular extrasystole were randomly divided into two groups: a treatment group, and a control group. In the control group, mexiletine was orally administered, and in the treatment group needling at bilateral Neiguan (PC 6) plus oral administration of mexiletine were given. 2 weeks of treatments constituted a therapeutic course. RESULTS: Of the 30 cases in the treatment group, 23 cases were markedly effective, 5 cases improved, and 2 cases failed; while in the control group 16 cases were markedly effective, 8 cases improved, 4 cases failed, and 2 cases aggravated. The total effective rates of 90.3% and 80.0%, respectively in the treatment and control groups showed a very significant difference between the two groups (P < 0.01). CONCLUSION: Needling at bilateral Neiguan (PC 6) may enhance the therapeutic effect for frequent ventricular extrasystole. PMID- 15119173 TI - What are the main points in acupuncture treatment of peripheral nervous injuries? PMID- 15119172 TI - Treatment of secondary amenorrhea with abdomen acupuncture. PMID- 15119174 TI - Ocular needling for treatment of sudden deafness in 40 cases. PMID- 15119175 TI - Acupuncture methods for treatment of hemiplegia. AB - There are several acupuncture methods for various types of paralyses, based on different selections of points and needling methods, i.e. needling the points selected by the yin-yang principle; needling the polar points; needling both the local and opposite points; needling the upper points with the lower ones needled as supplementary; and needling by the method of reinforcing the channels and reducing the collaterals. The specific application of each method are described below. PMID- 15119176 TI - Several problems in acupuncture treatment of headache. PMID- 15119177 TI - Acupuncture treatment of 3 difficult cases according to Dr. Xu Benren's experience. PMID- 15119178 TI - Prof. Zhou Yiqiang's experience in treatment of lung cancer. PMID- 15119179 TI - TCM treatment of lumbago due to qi stagnation in the liver and gallbladder. PMID- 15119180 TI - Effects of moxibustion at shenque (CV 8) on serum IL-12 level and NK cell activities in mice with transplanted tumor. AB - The study was to investigate the effect of moxibustion on the serum IL-2, IL-12 levels and NK cell and ascitic tumor cell activities in H-22 mice with tumor and its mechanisms. The method used in the study was that the IL-2 and IL-12 levels were determined by the double antibody method with ELISA, and the NK cell activities by the MTT measure. The results showed that moxibustion elevated the serum IL-2 and IL-12 levels and the NK cell activities of the mice with tumor, and the elevations were of significance as compared with the controls (P < 0.05). It is concluded that moxibustion can inhibit the growth of tumor, which is related to the increase of the serum IL-2 and IL-12 levels and the strengthening of NK cell activities. PMID- 15119181 TI - Effects of tang mai kang capsule on angioneurotic lesions in alloxan-induced diabetic mice. AB - The effects of Tang Mai Kang Capsule ([symbol: see text]) on blood sugar level, gangrene of the tail-tip, pain threshold and learning and memory abilities were investigated in alloxan-induced diabetic mice. The results showed that Tang Mai Kang Capsule could significantly decrease blood sugar level and incidence rate of gangrene of the tail-tip, increase pain threshold, and strengthen learning and memory abilities, suggesting that Tang Mai Kang Capsule functions to decrease blood sugar level and improve the complicated angioneurotic lesions of diabetes. PMID- 15119182 TI - Advances in diagnosis and treatment of hepatic steatosis. AB - Although many breakthroughs have been made in biomedicine, however, a satisfactory treatment is still lacking. On the other hand, treatment with TCM drugs has been proved satisfactory through there is no unified criterion on type differentiation in diagnostics. Hepatic steatosis due to alcoholic hepatitis could be effectively treated with the therapeutic methods of soothing the liver, promoting blood circulation and resolving phlegm. Generally speaking, hepatic steatosis can be treated with the principles of 1) soothing the liver, strengthening the spleen and resolving phlegm; 2) tonifying the liver and kidney, resolving phlegm and dispelling stasis; 3) soothing the liver and promoting blood circulation, strengthening the spleen and excreting dampness; 4) soothing the depressed liver to regulate the circulation of qi, softening hardness and promoting blood circulation; and 5) strengthening the spleen and excreting dampness and soothing the depressed liver to regulate the circulation of qi. PMID- 15119183 TI - How to treat pain in the hepatic region due to chronic hepatitis? PMID- 15119184 TI - Studies on TCM syndrome types of heroin-dependence. PMID- 15119185 TI - [Human brain evoked potentials and discrimination of short acoustic stimuli with various frequency]. AB - We found that the mismatch negativity (MMM) was absent in evoked potentials in passive condition for frequency deviant acoustical stimuli when stimulus duration was only 11-30 ms. But it was shown that it is possible for participants to fairly well discriminate these stimuli and the component N2b in RP was generated without preceding MMN. Processing negativity also was absent in potentials by the minimal stimulus duration (11 ms). Auditory discrimination nevertheless was still possible, but reaction time and number of committed errors increased significantly. PMID- 15119186 TI - [Significance of individual resistance to hypoxia for the correction of the brain trauma sequelae]. AB - In rats, the individual sensitivity to hypoxia plays an prominent role for recovery of animals after mechanic brain injury. Enthomerzol (25 mg/kg intraperitoneally) for three days after brain injury decreased behavioral disorders in rats with different resistance to acute hypoxia, recovered structure of individual behavior, prevented metabolic disorders in brain. Therefore, antihypoxant ethomerzol is effective as a drug against hypoxia due to brain injury. PMID- 15119187 TI - [Effect of alexithymia on the event-related theta-synchronization of human EEG during reception of emotional visual stimuli]. AB - The 62-channel EEG was recorded while control non-alexithymic (n = 21) and alexithymic (n = 20) participants viewed sequentially presented neutral, pleasant and unpleasant pictures and subjectively rated them after each presentation. The event-related synchronization (ERS) to these stimuli was assessed in the theta-1 (4-6 Hr) and theta-2 (6-8 Hz) frequency bands. The obtained findings indicate that alexithymia influences perception of only emotional stimuli. In the upper theta over anterior cortical regions alexithymia vs control individuals in response to both pleasant and unpleasant stimuli manifested decreased left hemisphere ERS in the early test period of 0-200 ms along with enhanced ERS in response to negative vs positive and neutral stimuli in the right hemisphere at 200-600 ms after stimulus onset. The findings provide the first EEG evidence that alexithymia construct, associated with a cognitive deficit in initial evaluation of emotion, is indexed by disrupted early frontal synchronization in the upper theta band that can be best interpreted to reflect disregulation during appraisal of emotional stimuli. PMID- 15119188 TI - [Effect of forced swimming on the memory track retention in mice with various behavioral stereotypes]. AB - The effects of forced swimming on retrieval of the passive avoidance during its extinction were found to depend on aggressive and submissive behavior. In mice without generated behavioral stereotypes, swim stress applied before or after training stabilized retention of the memory trace retrieval. The similar improved influence of forced swimming on memory storage is revealed in submissive, but not aggressive mice. The increase of resistance against extinction under the swim stress can be connected to facilitation of emotional processes. PMID- 15119189 TI - [Effect of a corticoliberin (CRF4-6) on the rat heart rate and behavior]. AB - The effects of tripeptide corticoliberin fragment CRF4-6 (Pro-Pro-Ile) on heart rate and behaviour of rats with simultaneous monitoring of these parameters in free-moving animals in their home cage were investigated. Intracerebroventriculary administered CRF4-6 (6, 30, 150 nmol) induces arousal effect increasing the duration of active behaviour, decreasing the duration of passive behaviour and sleep both. At the same time the tripeptide increases the heart rate during sleep, passive and exploratory activity. CRF4-6 (30, 150 nmol) also increases the heart rate in anaesthetised rats. All observed effects of the tripeptide are dose dependent. Taking into account these facts we suggest that CRF4-6 influences behaviour and heart activities independently. PMID- 15119190 TI - [Correlation between the organ blood flow, substrate absorption from blood, the activity of transport into mammary gland secretory cells and formation milk components in cow]. AB - Epithelial cells of mammary alveoles may he considered as a bioreactor that works being monitored by regulatory system maintaining balance between blood substrate supply and rate of macromolecular synthesis. In the trial performed on lactating cows assigned to feeding regimen with temporally altered level of nutrition, functioning of this system in the course of a transition period was studied. The earliest sign of adaptation was a decrease in volume blood flow through mammary gland provoking decline in uptake of water soluble substrates (glucose, amino N, P-hydroxybutyrate) with increase in arterio-venous difference across mammary gland and extraction efficiency. At the end of deprivation period, an activity of transport into the cell decreased for amino N from 7.5 to 4.5; l/min (p < 0.05), for P-hydroxyhutyrate from 16.0 to 13.8 l/min; activity of glucose transport was not changed. The data obtained indicate existence of defined points in the system monitoring organ blood supply and transport of substrates into the cell being adjusted during adaptation to alteration in the level of nutrition. PMID- 15119191 TI - [Mechanical work of the left ventricle of the human heart]. AB - By means of the echocardiographic techniques, morphometric and functional parameters of the heart left ventricle (HLV) were studied in male humans aged 20 to 23, in resting and under the effect of a physical load. The total ventricle work was found to be about 1 J, the relative one (per 100 g of the LV mass)--0.6 J. The total working power was found to be about 70 Watt, the relative one--about 40 Watt. The physical load resulted in increase of the LV mechanical work parameters due to the heart rate and systolic arterial pressure. PMID- 15119192 TI - [Switching off the estrogen effect and approaches to its correction]. AB - Data are summarized on experimental means of induction of PSEE and its modification/prevention with the aim to achieve optimal ratio of estrogenic effects (as low as possible genotoxicity in combination with satisfactory and excessive hormonal action). Among studied agents were ethanol, tobacco smoke, irradiation, aging (as PSEE inductors) and carnosine, N-acetylcystein, vit. E and C, melatonin, swimming and antiestrogen ICI 182780 (as PSEE modificators). PMID- 15119193 TI - [Characteristics of the K(+)-ion transport in the rat intestine following the use of natural zeolites as food additives]. AB - Effects of zeolites as a food supplement have been studied on Wistar rats both in vivo perfusion experiments in the jejunum and distal colon and Rb fluxes through intestinal wall in the Ussing chamber. It has been found that zeolites decrease the K+ absorption and stimulate K+ secretion in the gut. This effect was due to inhibition of the apical N(+)-K(+)-ATPase and ouabain-sensitive Na(+)-independent K(+)-ATPase as well as the activation of the basolateral N(+)-K(+)-ATPase. PMID- 15119194 TI - [Involvement of kappa-opioid receptors in mechanisms of aggressive and submissive types of behavior in male mice]. AB - Effects of the kappa-opioid receptor agonist U-50.488H (0.0, 0.6, 1.25, 2.5 mg/kg. s.c., 30 min) on behavior of the winner with repeated experience of victories and the losers with repeated experience of social defeat in 20 daily agonistic confrontations as well as the control mice were investigated in the tests estimating exploratory activity (open-field) and communication (partition test). Different effects of drug on behaviors of animals with different social story were shown in both tests. In the losers, all doses of U-50.488H had anxiolytic effect, increasing the communication in the partition test. In the winners, the drug induced an increase of aggressive motivation. The control mice were less sensitive to the treatment. In the open-field test, U-50.488H increased the locomotor and exploratory activity in high anxious losers. Winners significantly differed in their reaction to drug treatment in most behavioral forms in comparison with the controls and losers. It was concluded that kappa opioid receptors are specifically involved into mechanisms of formation of aggressive or submissive types of behaviors under positive or negative social experience. PMID- 15119195 TI - [Effect of the D1-receptor antagonist SCH-23390 on the individual and aggressive behavior in male mice with various aggression experience]. AB - It has been shown that dopaminergic systems are involved in mechanisms of aggressive behavior. Effects of SCH 23390 (dopamine Di receptors antagonist. 0-1 mg/kg, i/p, 30 min) on aggressive and individual behaviors were studied in male C57BL/6J mice with different experience of aggression. SCH 23390 reduced aggressive attacks in animals without preliminary experience of aggression. However total time of hostile behavior (sum of the total time of attacks, aggressive grooming and diggings) didn't changed. No significant effects on behaviors were found in mice with long (20 days) repeated experience of aggression. It was supposed that long aggressive experience produces pharmacological desensitization of Di receptors as a result of enhanced dopaminergic activity shown earlier in aggressive animals. PMID- 15119196 TI - [Role of catecholamines in the glucose absorption regulation in the small intestine]. AB - During the experiments in vivo at white laboratory rats-males Wistar was established that the epinephrine and norepinephrine depending on the dosation causes the stimulation or inhibition of frequency of glucose absorption in the small intestine. The stimulating effect of epinephrine is mediated beta-, inhibitory--as alpha 1- and alpha 2-adrenoreceptors; the potentive effect of norepinephrine--by alpha-, inhibitory--alpha 1--adrenoreceprors. It was found out the personal physiological role of dopamine as inhibitory mediator for absorption of glucose in the small intestine. PMID- 15119197 TI - [Effect of various methods of oxygenation in the isolated small intestine on the tryptophan and glycine accumulation]. AB - The impact of various oxygenation types of intestinal preparations (mucosal, serosal and bilateral) on the transepithelial transport of amino acids and peptide according to the character of amino acids, their stereoisometry, incubation time, intestinal gradient and other factors, was investigated in fowl. Against-the-gradient-transport of L-tryptophan and glycine was observed at various oxygenation types, and its intensity decreased in the following sequence: bilateral, mucosal, serosal oxygenation. At various oxygenation types, the accumulation of L-tryptophan occurs more efficiently than that of DL-tryptophan and D-tryptophan. Using an in vitro bilateral intestinal oxygenation model system, we have demonstrated the plasma membrane of enterocyte microvilli to be rapidly impaired after oxidative stress. Glycine, but not L-tryptophan, attenuates oxidative injury in brush border membrane and alterations in amino acid transport activity. Overall, our data indicates that in vitro serosal oxygenation of the duodenum markedly improves glycine absorption, possibly involving the basolateral transporters. PMID- 15119198 TI - [Indirect evaluation of the transport activity of metabolites into cells in vivo based on measurements of their arteriovenous balance]. PMID- 15119199 TI - [Chronic renal insufficiency: refer in time]. AB - Three patients reaching end-stage renal failure were referred to an internist nephrologist. A 53-year old woman was referred late, with several late complications of chronic renal failure such as secondary hyperparathyroidism and signs of left ventricular hypertrophy. In addition she suffered from complications of a temporary vascular access required for intermittent haemodialysis prior to commencing peritoneal dialysis. A 52-year old man was referred early, resulting in reduced late complications of chronic renal failure and the timely start of peritoneal dialysis. An 84-year old man was referred for optimization of metabolic control and to be informed about the possibilities and consequences of renal replacement therapy, which he subsequently refused. With early consultation or referral to a nephrologist (i.e. when the creatinine clearance is < 30 ml/min) complications associated with chronic renal insufficiency can be identified and treated early, with the intention of preventing or reducing their impact, resulting in reduced morbidity and death, even after starting dialysis treatment. PMID- 15119200 TI - [The practice guideline 'Acute cough' from the Dutch College of General Practitioners; a response from the perspective of general practice]. AB - The practice guideline 'Acute cough' from the Dutch College of General Practitioners stresses the fact that a cough of less than 3 weeks' duration seldom heralds serious pathology. However, for sound reassurance of patients presenting with a cough of short duration, the general practitioner needs to know much about the signs and symptoms connected to low-prevalence serious pathology in these patients. The practice guideline distinguishes upper and lower respiratory tract infections and defines serious lower respiratory tract infection. The diagnostic value of symptoms and laboratory findings like a sedimentation rate or C-reactive protein in order to make such distinctions, is not explained in detail. Antibiotics are reserved for serious lower respiratory tract infection with the exception of acute bronchitis, croup and bronchiolitis, which can be treated without antibiotics. Recommendations for treatment of acute bronchiolitis with bronchodilators or corticosteroids, and croup with corticosteroids are based on consensus. This practice guideline can be considered as a clear and valuable piece of work for all physicians in primary and secondary care. PMID- 15119201 TI - [The practice guideline 'Acute cough' from the Dutch College of General Practitioners; a response from the perspective of pulmonology]. AB - Coughing is one of the most prevalent symptoms for which patients seek medical attention. Acute cough is defined as a symptom that lasts less than 3 weeks, is mostly transient and with minor consequences. Upper airway infections, and especially the common cold, are the main cause of acute cough. Acute cough is only occasionally due to life-threatening causes like severe pneumonia or pulmonary embolism. The Dutch College of General Practitioners has issued a comprehensive, practical guideline for diagnosis and treatment of acute cough. As viral infections are the most prevalent cause of acute cough, the guideline emphasises the need to be as cautious as possible with the prescription of antibiotics. Unfortunately, a paragraph on the causes of reduced cough effectiveness, which may lead to a more complicated course of illness, is missing from this guideline. Hopefully, this guideline will lead to a more standardized approach to patients presenting with acute cough. PMID- 15119202 TI - [Prevention demands a more ambitious approach. Response to the cabinet policy document 'Live healthier for longer 2004-2007; also a matter of healthy behavior']. AB - In the cabinet policy document 'Live healthily for longer 2004-2007; also a matter of healthy behaviour' the Dutch Secretary of State for Health, Welfare and Sport assumes that many health problems are linked to behaviours such as smoking and lack of physical activity. He places the prime responsibility for that with the citizens and also with local councils, trade and industry, care providers and schools. However conscious behavioural choices by individual citizens form only a small part of the cause-effect chain that leads to ill health. It is incorrect to ascribe the differences in unhealthy behaviour to freely made behavioural choices. Moreover it begs the question as to whether the proposed measures will be effective. Over and above all this, the finances that are available to strengthen preventative measures are completely inadequate. The central government should be encouraging the health care sector to devise a more central position for prevention and ensuring that effective opportunities for prevention are used as much as possible. After all, it is expected of the central government that it will look after the collective interests and protect and promote the health of the population. The necessary improvements can only be achieved through a much more ambitious approach than that presented by the cabinet. PMID- 15119203 TI - [Nutrition and health--nutrition and performance in sports]. AB - Optimal nutrition is an important prerequisite for a top-level performance in sports. Of primary importance in this connection is the bioavailability of carbohydrates and fats as a source of energy so that the muscles can produce ATP. The amount of glycogen in the liver and skeletal muscles and possibly the intramuscular triglyceride levels play a pivotal role in this process. Gradually decreasing the amount of training during the last 6-7 days before an important game and simultaneously increasing the amount of dietary carbohydrates results in a higher physical performance. Studies have shown that carbohydrate-electrolyte solutions with 6-8% carbohydrate by weight and an osmolarity of 250-350 mosmol/l result in a maximal carbohydrate oxidation from the solution of about 1.0-1.1 g/min and a significantly enhanced endurance. A combination of carbohydrates and proteins or amino acids has proven to be very effective to accelerate recovery after exhausting exercise. Functional food ingredients such as caffeine and creatine result in a significantly enhanced performance. This is contrast to many other ingredients and products for which the extravagant claims have not been substantiated. Hence, in addition to a few functional sports supplements for which the value has been scientifically demonstrated, sound nutrition remains an absolute prerequisite for an optimal performance in sports. PMID- 15119204 TI - [Podocyte dysfunction and proteinuria]. AB - Podocytes play a central role in the pathogenesis of several glomerular diseases. In recent years, this has been revealed by molecular analysis of a number of rare hereditary renal diseases. Podocytes contain three domains: the domain bound to the glomerular basement membrane (GBM), the domain of the slit diaphragms and the apical domain. The slit diaphragms are situated basolaterally between the pedicles and form together with the GBM a mechanism for the selective filtration of blood to primary urine. The apical cell membrane forms a negatively charged layer which prevents adhesion to the adjacent cell membranes, thus keeping the slit diaphragms and urinary space open. Many podocyte diseases are characterised by foot process effacement, which causes the loss of slit diaphragms, and could lead to podocyte loss. Specific abnormalities have been discovered in the three domains of the podocyte to which a number of glomerular diseases can be attributed. PMID- 15119205 TI - [Treatment of patients with chronic renal insufficiency; a guideline for internists]. AB - Early optimalization of the treatment of patients with chronic renal insufficiency can reduce morbidity and deaths. For each patient with a raised serum creatinine concentration, the creatinine clearance should be measured or calculated. When this is abnormal, the cause thereof should be investigated. Chronic renal insufficiency is often progressive, even when the initiating factors are no longer present. Progression can be delayed by treating the high blood pressure, proteinuria and hyperlipidemia by means of a restricted protein diet and advice not to smoke. Acute deterioration of an existing chronic renal dysfunction through dehydration and underfill or through the use of certain medications or toxic substances such as radio-opaque media should be avoided. In patients with chronic renal insufficiency specific attention should be paid not only to hypertension, lipid disturbances, smoking and weight, but also to the calcium-phosphate balance, anaemia and homocysteine levels. The blood pressure, oedema and weight of patients with a clearance between 30-59 ml/min should be checked 2-3 times a year, in addition to laboratory tests for Hb, Ht, creatinine, urea, potassium, calcium, phosphate, pH, bicarbonate and lipid spectrum. It is recommended that when creatinine clearance (< 50 ml/min) falls a nephrologist should be consulted at least once with respect to the strategy to be followed. Symptoms of chronic renal insufficiency can occur when the creatinine clearance is < 30 ml/min. This relates to: sodium retention, imbalances in the calcium and phosphate levels, anaemia, uraemia, water retention, potassium retention and metabolic acidosis. Referral should take place at a creatinine clearance of < or = 30 ml/min. PMID- 15119206 TI - [Summary of the practice guideline 'Acute cough' from the Dutch College of General Practitioners]. AB - In most cases acute cough has an infectious, often viral, cause. When the coughing lasts longer than 3 weeks, the diagnosis has to be reconsidered. The effectiveness of cough medicines has not been proven. For the management of acute cough it is important to distinguish between non-serious and serious lower respiratory tract infections. A serious lower respiratory tract infection is: a lower respiratory tract infection with a higher risk of a complicated course: when pneumonia is suspected, in infants and the elderly, and in patients with relevant co-morbidity. The prescription of antibiotics for acute cough is not useful in the majority of the patients; antimicrobial therapy can be indicated, but only in the case of lower respiratory tract infections with a higher risk of a complicated course. Specific management has to be considered in the case of: whooping cough, bronchiolitis and croup. In the case of pneumonia, antimicrobial therapy is indicated; follow-up is necessary. In the case of moderate to severe croup, a single dose of corticosteroids is recommended. PMID- 15119207 TI - [Diagnostic image (184). Two women with painful bands after axillary lymph node removal. Lymphatic thrombosis and fibrosis]. AB - Two women aged 49 and 54 years, respectively, developed painful taut bands in their shoulder and elbow area after axillary lymph node dissection. These bands appear to be thrombosed and fibrosed lymphatic vessels. PMID- 15119208 TI - [Extent and quality of anti-coagulation treatment with coumarin derivatives by the Dutch Thrombosis Services]. AB - OBJECTIVE: To obtain an impression of the extent and quality of the anti coagulation treatment with coumarin derivatives carried out by the Thrombosis Services in the Netherlands. DESIGN: Descriptive. METHOD: Data were drawn from the medical annual reports of 62 of the 63 Thrombosis Services in the Netherlands over the period 1998-2002. In 2002 the Thrombosis Services treated 325,072 patients and performed 4,469,730 INR laboratory tests. The half-yearly figures produced by the Thrombosis Services were calculated as an average percentage per year per thrombosis service and then recalculated as a percentage per year. RESULTS: Seventy-three per cent of the patients were treated for an arterial and 27% for a venous indication. Depending on the required intensity of anticoagulation a mean of 74-78% of the long-term treated patients fell within the therapeutic range and a mean of 6-10% below. The mean number of major bleedings per 100 treatment years was 1.0. A mean of 79% of the patients was treated with acenocoumarol and 21% with phenprocoumon. When acenocoumarol was used, a mean of 72-77% fell within the therapeutic range and in the case of phenprocoumon 79-82%. In the last few years the number of patients had increased due to a growing number of patients treated for atrial fibrillation. The percentages of INR within the therapeutic range were unchanged or showed a slight increase. CONCLUSION: The quality of the anticoagulation therapy with coumarin derivatives was good or acceptable. PMID- 15119209 TI - [Spontaneous rupture of the bladder]. AB - A spontaneous bladder rupture was diagnosed using ultrasound and CT scan in 3 patients, 1 woman aged 62 and 2 men aged 77 and 42, presenting with abdominal pain, anuria and prior problems with urination. The younger man had suffered from this previously; he was now suffering from bacterial peritonitis due to infected urine. All 3 patients recovered after treatment with a transurethral catheter and antibiotics. A transurethral resection was performed on the benign enlarged prostate of the older man. The combination of non-specific symptoms, the absence of trauma history and its very rare occurrence mean that initially a spontaneous bladder rupture may not be suspected. However, an untreated bladder rupture can lead to life-threatening situations. Screening the abdomen with ultrasound, an ultrasound-guided puncture and CT scan may yield results that suggest a bladder rupture. In most cases, conservative treatment with a transurethral catheter will be sufficient. Sometimes surgical repair of a bladder rupture may be necessary. PMID- 15119210 TI - [Intestinal incarceration due to a Bochdalek's hernia]. AB - A 67-year-old male presented with abdominal pain followed by respiratory distress. Imaging revealed a right-sided Bochdalek's hernia. At thoracotomy two days later, an incarcerated small bowel segment was removed, with a protracted post-operative course. In adults, a Bochdalek's hernia is rare and usually asymptomatic. However, when symptoms do occur, incarceration of the intestinal contents is frequent and accompanied by high mortality. The severely protracted course after delayed treatment of a symptomatic Bochdalek's hernia described here illustrates the importance of early recognition and surgical intervention. PMID- 15119211 TI - [Retention period for patient data: the health council of the Netherlands pleads for a change in the law]. AB - According to the Dutch Medical Treatment Act (WGBO), patient data must be retained for at least 10 years or as much longer as required for the provision of good medical care. In practice, patient data are often destroyed after 10 years. This may lead to problems, for example if a treatment turns out to have long-term consequences. A committee of the Health Council of the Netherlands has now recommended a change in the law. They propose a longer retention period for the care of the patient, a statutory regulation governing retention for research purposes and supplementary provisions that make it possible to retain hereditary data in the interest of relatives. Until the necessary changes in the law have been made, further destruction of medical data must be prevented. This can be accomplished by revising the existing transition provision in the WGBO, while extending it by 5 years. The scientific societies can use this time to develop guidelines for the longer storage of data in specific areas of care and for the periodic cleaning up of the files. PMID- 15119212 TI - [Infant crying diary: a useful aid in distinguishing between normal and excessive crying behavior]. PMID- 15119213 TI - [Infant crying diary: a useful aid in distinguishing between normal and excessive crying behavior]. PMID- 15119214 TI - [Primary venous mesenterial thrombosis in three patients]. PMID- 15119215 TI - [Percutaneous dilating tracheostomy in intensive-care patients: technique, indications and complications]. PMID- 15119216 TI - [Percutaneous dilating tracheostomy in intensive-care patients: technique, indications and complications]. PMID- 15119217 TI - [Reasons why patients do or do not participate in clinical trials; a systemic review of the literature]. PMID- 15119218 TI - [Early information: a necessity for chronic renal insufficiency]. PMID- 15119219 TI - [Nocardiosis--is it frequently observed after the introduction of new immunosuppressive agents in renal transplantation?]. AB - In our series of 1374 renal transplantations performed between February 1970 and December 2002, we observed 6 cases of infection due to Nocardia asteroides. There were 4 males and 2 females, aged 49.8 +/- 12 years (29 to 63 years). One patient received his first transplantation and the 5 others retransplants. Three patients had PRA > 80%, one 28% and one 40%. One patient was diabetic and two had HCV infection. Two of 6 patients experienced acute rejection episodes. Nocardiosis localisation was pulmonary in 5 cases, cerebral in two and mediastinal in one. All patients recovered after reduction of immunosuppression and appropriate antibiotherapy with trimethoprim-sulfamethoxasole (TMP-SMX). When we analyzed the role of immunosuppression, we observed that only two cases were observed in the 933 recipients transplanted between 1985 and 2002 and receiving cyclosporin, contrasting with 4 cases among 174 recipients transplanted between 1996 and 2002 and receiving tacrolimus. Our data suggest that high immunologic risk patients, heavy immunosuppression, and perhaps tacrolimus-based immunosuppression are risk factors of nocardial infection. Early diagnosis of this severe infection, reduction of immunosuppression and appropriate therapy with TMP-SMX resulted in complete recovery in all our patients. PMID- 15119220 TI - [Hemolytic-uremic syndrome secondary to scorpion envenomation (apropos of 2 cases)]. AB - HUS was recently described following scorpion sting. We report 2 cases of HUS in the intensive care unit of a university hospital. Two children aged respectively 10 months and 1 year were admitted in the ICU after severe scorpion envenomation (with coma and pulmonary oedema) having required dobutamine and mechanical ventilation. Evolution was marked with acute anaemia without bleeding requiring blood transfusion, acute renal failure, low platelets and signs of haemolysis. Our experience and the previously reported case suggest that scorpion sting could be added to the list of causes of the HUS. PMID- 15119221 TI - [A rare cause of pulmonary-renal syndrome with autoantibodies: primary antiphospholipid syndrome]. AB - We present a case of primary antiphospholipid syndrome with acute renal failure and alveolar haemorrhage. He was successfully treated with cyclophosphamide, corticosteroids and plasma exchange. Patients with antiphospholipid syndrome may develop a broad spectrum of pulmonary disease. Pulmonary thromboembolism and pulmonary hypertension are the most common complications, but alveolar haemorrhage have also been reported. Other causes need to be excluded. Despite favourable outcome of many patients with association of plasma exchange, immunosuppressive drugs and anticoagulant therapy, definite conclusions about the best therapeutic regimen could not be draw. PMID- 15119222 TI - [Extramembranous glomerulonephritis and myelodysplastic syndrome]. AB - Membranous glomerulonephritis (MGN) is the main cause of nephrotic syndrome in adults and is usually idiopathic. We report a case of nephrotic MGN associated with a myelodysplastic syndrome (MDS) in a 43 year old man. The initial treatment consisted of oral corticosteroids (1 mg/kg/day). Within 3 months proteinuria decreased from 22.4 g/day to 3.96 g/day and the blood cell count was normalized. Renal biopsy disclosed type I MGN. Ponticelli's protocol was started with a favorable effect: negative proteinuria, normal blood cell count and normal bone marrow cellularity. The association between MGN and MDS is quite rate. The possible links between the two conditions are reviewed. PMID- 15119223 TI - [From speeches to action]. PMID- 15119224 TI - [Wheat to celiac disease]. PMID- 15119225 TI - [Screening for lung cancer using low dosage computer tomography]. PMID- 15119226 TI - [SARS, drugs and direction of Golf-stream. Globalization influences everything, including health]. PMID- 15119227 TI - [Does a doctor care about a cancer patient's pain?]. PMID- 15119228 TI - [Bioabsorbable devices in bone surgery]. PMID- 15119229 TI - [Secrets of sunscreens]. PMID- 15119230 TI - [Asperger syndrome in adults]. PMID- 15119231 TI - [Sudden deterioration and later improvement of an elderly dementia patient]. PMID- 15119232 TI - [Management of severe synovial arthritis with drugs]. PMID- 15119233 TI - [Obesity in adults]. PMID- 15119234 TI - [Absorption and synthesis of cholesterol in patients with type 1 and type2 diabetes mellitus]. PMID- 15119235 TI - [Young women suffering from shortness of breath]. PMID- 15119236 TI - [About the portrait of Johan Wilhelm Runeberg]. PMID- 15119237 TI - [Depression. Practice guideline set up by the Society of Finnish Psychiatrists]. PMID- 15119238 TI - What potential risks might I be exposed to working with anaesthetic gases? PMID- 15119239 TI - All things being equal.... AB - As most of us are aware, Agenda for Change is the new terms and conditions package for all NHS staff. Although it is currently being trialled at designated early implementer (EI) sites in England and Scotland, the agreement has been accepted by the Health departments of all four home countries of the UK. It is to be introduced to all NHS staff in October 2004. PMID- 15119240 TI - Clinical supervision. AB - The NHS Plan (DoH 2000) states that the modern National Health Service (NHS) will be driven by a cycle of continuous quality improvement. This will enhance the efficiency, productivity and performance of the NHS. The aim of this article is to examine clinical supervision as a quality assurance initiative. To achieve this, clinical governance and lifelong learning will be evaluated within a theoretical framework with the focus on clinical supervision. PMID- 15119241 TI - The role of the first assistant in robotic assisted surgery. AB - In part one of this series (BJPN 14 (1) 36-39) the introduction of the da Vinci robotic system was described (Intuitive Surgical, Inc, Mountain View, CA). This second and final part in the series describes the unique role of the first assistant in robotic assisted surgery. In September 2000, the da Vinci robotic system, the first of its kind to be installed in the UK, was introduced to St Mary's Hospital, London. PMID- 15119242 TI - Management of a CJD case. Part 1. Preoperative organisation of the case. AB - Recent years have seen the rise of variant Creutzfeldt-Jacob Disease (vCJD), causing great concern amongst the public at large and healthcare professionals, and the implications of the disease are still to be properly determined. In this, the first of two articles, Beverley McNeil outlines the various risk categories of CJD patients, the planning of the case and the handling of associated instrumentation postoperatively. Next month's article will deal with the practicalities surrounding management of the case. PMID- 15119243 TI - Work-life balance. AB - With the National Midwifery Council reporting that numbers of UK Midwives are at the lowest for ten years, it seems reasonable to question whether this is 'putting lives at risk'. PMID- 15119244 TI - Gravitational platelet separation. Accelerating the body's own healing process. PMID- 15119245 TI - Procreative liberty in the era of genomics. PMID- 15119246 TI - Fool's gold: psychologists using disingenuous reasoning to mislead legislatures into granting psychologists prescriptive authority. PMID- 15119247 TI - Fake pharmaceuticals: how they and relevant legislation or lack thereof contribute to consistently high and increasing drug prices. PMID- 15119248 TI - Picking up the pieces. PMID- 15119249 TI - A nurse at heart. PMID- 15119250 TI - Can we cope? Bioterrorism and the NHS. PMID- 15119251 TI - Scaphoid fractures. PMID- 15119252 TI - Pharmacological techniques for managing acute pain in emergency departments. AB - Pain is a common reason to attend hospital but it is often inadequately managed. This article examines the physiology of pain and suggests the most effective pharmacological techniques to control it. PMID- 15119253 TI - Should smoking be outlawed? PMID- 15119254 TI - Bullying and nursing. PMID- 15119255 TI - Promoting respect and dignity. PMID- 15119256 TI - The nurses improving sexual health. PMID- 15119257 TI - Self-harm: understanding the causes and treatment options. AB - Deliberate self-harm rates in the UK are the highest in Europe, and at present, this problem is poorly understood, as there has been little research available to shed light on why such a considerable number of young people are compelled to hurt themselves. The Mental Health Foundation and The Camelot Foundation recently launched a UK-wide inquiry into self-harm among 11-25-year-olds, which aims to produce a practical and thoughtful approach to the prevention and treatment of self-harm in young people, inform future education and training, and provide strategies for service providers. PMID- 15119258 TI - What you need to know about ... chickenpox. PMID- 15119259 TI - The development of best practice in breaking bad news to patients. AB - There can be few tasks that a health care practitioner has to undertake that impact more upon patients and their relatives, than that of breaking bad news. Consideration needs to be given to the people involved, the communication process, and the environment. This article highlights the issues that make for best practice in this important area of care--including who gives the information, how it is presented, and the nature of the environment where it is given. PMID- 15119260 TI - A trust-wide strategy for the management of anaphylaxis. AB - Anaphylaxis is a life-threatening medical emergency. Although it is uncommon, when it does occur prompt recognition and appropriate treatment are essential. This article describes how a multidisciplinary working party review of practices concerning the management of anaphylaxis in an acute hospital trust led to the introduction of standardised anaphylaxis emergency boxes. PMID- 15119261 TI - Aetiology, signs, symptoms and treatment of trigeminal neuralgia. AB - Trigeminal neuralgia is one of the most common causes of facial pain. It can be excruciating and can severely incapacitate patients and negatively affect their quality of life. First-line treatment is anticonvulsant drugs but other pharmacological options are available. If drug therapy is ineffective, a range of surgical options exist. PMID- 15119262 TI - A role in changing public attitudes. PMID- 15119263 TI - Child-specific specialties increase. PMID- 15119264 TI - [Changing trends in the treatment of nodular thyroid disease--own experience]. AB - If the first surgical intervention for benign euthyroid goiter diseased portion is not totally removed one cannot be certain of the long-term behaviour of the remaining tissue. In one third of these patients there may be a recurrence with new nodules developing, functional disorders or even carcinoma may appear in the remnants. Hormonal activity of the removed tissue can be completely substituted for by levothyroxine. That is why some experienced specialists recommend primarily radical surgery even in benign disease, based on their experience with large number of patients. The authors started performing radical excisions in the past three years. A total of 477 operations were performed between 1st January 2000 and 31st December 2002, 45% were radical: lobectomy, near-total thyroidectomy and bilateral lobectomy. Surgery was carried out according to the standardized morphologically adjusted technique (SMART). Results of these operations are compared to conventional procedures performed in the same period. Complication rate was low with no mortality. There was no significant difference between early results of the two groups. About 2% rate of permanent recurrent laryngeal nerve injury a 3% rate of postoperative tetany (hypocalcaemia) in both groups is acceptable. Based on their initial experiences the authors recommend the radical surgical technique for the experienced surgeons. PMID- 15119265 TI - [Intraoperative ultrasonography for common bile duct exploration during laparoscopic cholecystectomy]. AB - The "Endomedix Laparoscan" and the "Leopard" and "Panther" intraoperative ultrasounds were successfully used for the detection of unsuspected common bile duct stones during laparoscopic cholecystectomy (LC). Out of 60 patients six had common bile duct (CBD) stones and in one patient sludge has been seen. In patients with CBD stones, four small calculi have been observed in one patient, despite negative intraoperative cholangiography (IC). In an other patient a stone in the retropancreatic part of the CBD was detected. Based on preoperative findings CBD stone was unsuspected. We found that intraoperative ultrasound (IOUS) is useful for in investigating the CBD to detect unsuspected common bile duct stones. It can be used for the examination of other organs (liver, pancreas, hepatoduodenal ligament) as well. The method is easy to perform, fairly simple and informative so it can replace IC during laparoscopic cholecystectomy. PMID- 15119266 TI - Videomediastinoscopy for the diagnosis of the diseases of the lung and mediastinum. AB - BACKGROUND: The biopsy of the mediastinum by mediastinoscopy is established as the most accurate method especially in the lung cancer staging, but helps also in the diagnosis of various diseases of the lung and mediastinum. The videomediastinoscopic equipment offers more precise procedure and shorter learning curve in comparison with classical technique due to the sharing of the procedure with other (more experienced) surgeon. The aim of our work was to confirm the safety and reliability of the videomediastionoscopy in our department, where the use of the traditional technique was exceptional. METHODS: Patients with expansion in the mediastinum (or enlargement of the mediastinal lymph nodes) of the unknown aetiology accessible by videomediastinoscopy according to CT were included. Standard videomediastinoscopic technique was used. The attempt to take biopsy of the mediastinal tumor and/or the lymph nodes in stations 1, 2, 4, 7 according the UICC regional lymph node mapping was done. RESULTS: There were 126 patients operated by videomediastinoscopy between January 1998 and June 2003. There were 83 men with a mean age of 50.43 years (range 14 to 78 years) and 43 women with a mean age of 50.79 years (range 18 to 77 years). The diagnosis defined by explicit morphology was made in 95 cases (75.40%). In 25 cases (19.84%) the result of the biopsy was either normal lymphatic tissue or slight inflammatory changes. In three cases (2.38%) the pathologist expressed strong suspicion of the malignant disease, without clear morphology. In two cases (1.59%) all tissue samples were completely necrotic. No biopsy was done once (0.77%). There were four major groups of patients with positive histology: Patients with sarcoidosis, patients with metastases of the epithelial malignity to the mediastinal lymph nodes, patients with lymphoma and the group containing patients with a diagnosis of a low frequency (tuberculosis, sarcoma, Castleman's disease and others). No mortality was related to the procedure. We had one serious surgical complication: bleeding from the right innominate artery successfully treated by suture. CONCLUSION: Videomediastinoscopy is a safe and accurate method with a short learning curve. The key point of the diagnostic success is a proper indication for the procedure. Videomediastinoscopic equipment and its use is to be recommend to each department of general thoracic surgery. PMID- 15119267 TI - [Modified Jatene operation for the repair of left ventricle aneurysm]. AB - INTRODUCTION: 134 left ventricle aneurysm operations were performed at our institute between 1993 and 2000. The original Jatene operation was modified by our workgroup: running sutures and abandoning Teflon pledges for faster operation; an additional linear closure of the remnant aneurysm above the patch to eliminate bleeding complications. OBJECT: The aim of this study was to describe our surgical technique, its advantages against other surgical methods in left ventricle aneurysms. METHOD: We divided our patients into three subgroups: linear techniques, such as plication (PL) n = 54, resection (RE) n = 45, and circular technique modified Jatene operation (MJ) n = 35. We performed a I. retrospective study in these subgroup, and a II. long-term clinical follow-up in MJ group. RESULTS: We did not find any technical difficulties in modified Jatene operations (there was no significant difference in operating times and cross clamping times in linear versus circular techniques). On the other hand, we found significantly better left ventricle geometry after circular method with echocardiography. There was no short-term (< 30 days) mortality in RM group. II. In the RM group long-term clinical follow-up we found further improvement in left ventricle geometry. We lost 1 patient (2.94%) because of cardiac death, and 2 patients (5.88%) had heart transplantation. All patients were in NYHA class I or II. CONCLUSIONS: In left ventricle aneurysm surgery circular techniques should be preferred because the better results in left ventricle geometry. We think that our improvements in Jatene's operation significantly decreased surgical difficulties, and through decreasing operating time short and long term outcome can be positively affected. PMID- 15119268 TI - [Surgical treatment of malignant renal tumors invading the inferior vena cava and right atrium]. AB - Invading the inferior vena cava and right atrium is the most serious, but fortunately not common complication of renal cell carcinoma. Radical nephrectomy with tumor-thrombus extraction is the only way to improve these patients survival. Cardiopulmonary bypass with or without deep hypothermia and total circulatory arrest might be necessary during surgery. Between 1998 and 2003 at the Department of Cardiac Surgery of University of Debrecen, 5 patients, with renal cell carcinoma extending into the right atrium, had radical nephrectomy and thrombectomy. We used cardiopulmonary bypass, in 2 patients in total circulatory arrest, in deep hypothermia. There was no operative death and neurological complications. One patient died 3 years after the operation due to cardiac failure. In average 42 months after surgery, 4 surviving patients are under regular follow up, they have a good quality of life, without recurrence. In our opinion cardiopulmonary bypass and total circulatory arrest, if necessary, gives the best way for surgical resection of renal cell carcinoma extending into the right atrium. PMID- 15119269 TI - [Uncommon case of incomplete left paraduodenal hernia]. AB - Internal hernias are very rare. The hernial orifice, sac and content are situated inside the abdomen. Paraduodenal hernias are relatively rare congenital malformations and account about 50 per cent of all internal hernias. The cause of this malformation is the incomplete rotation of the mid-gut. Right and left paraduodenal hernias are different, varying in anatomic structure and embryological origin. In right paraduodenal hernia the small bowel is partially or completely localised behind the mesocolon of the ascending colon, in left paraduodenal hernia behind the mesocolon of the descending colon. That is why the widely used name "mesocolic" hernia is more convenient, because it refers on the pathogenesis of the disorder. The complaints can vary from recurrent atypical abdominal pains to the complete small bowel obstruction, but often there are no complaints. Abdominal CT scan and the barium meal provide the best diagnostic approach for paraduodenal hernias, but it will be recognised very often only at an emergency operation. A case of small bowel obstruction caused by incomplete left paraduodenal hernia discovered after swallowing a foreign body is described with pathogenesis, diagnosis and possible treatments for the disease. PMID- 15119270 TI - Combined off-pump coronary artery by-pass and right upper lobectomy for lung cancer through median sternotomy. AB - Patients who smoke are at risk of both coronary artery disease and lung cancer, but concomitant lesions of the heart and lung are uncommon. Management of these concomitant lesions is controversial because of the systemic side-effects of a cardiopulmonary by-pass (CPB) and the difficult approach of the lung. An off-pump coronary artery by-pass (OPCAB) can be a successful procedure in the combined treatment of lung cancer and cardiac disease. This study reports on a successful one-stage OPCAB grafting procedure with right upper lobe lobectomy for a primary lung cancer. PMID- 15119271 TI - [To the Editor in Chief]. PMID- 15119272 TI - HIV and host genetics. Complexity and contradiction. PMID- 15119273 TI - Higher Sustiva levels seen in some African-Americans. PMID- 15119274 TI - Best of the retrovirus webcasts. PMID- 15119275 TI - World CAB. Focus on international drug pricing. PMID- 15119276 TI - News from the bench. Resistance to RNAi inhibition of HIV. PMID- 15119277 TI - News from the bench. Assay for less-fit phenotype? PMID- 15119278 TI - Doctors organize to protest Abbott's Norvir price hike. PMID- 15119279 TI - Boosted Reyataz: 48-week results. PMID- 15119280 TI - Fortress NIH. PMID- 15119281 TI - A call to action: strategies for effective advocacy in challenging times. PMID- 15119282 TI - General health maintenance strategies. AB - Have you ever heard someone say, "I don't want HIV to take over my life. My life is more than my HIV status?" Perhaps you've had similar feelings or felt overwhelmed with trying to manage your health. This article provides a different way of thinking about health than what many people may experience at a doctor's office. The goal is to offer a framework for thinking about a big picture of well being and provide a path for developing a long-term strategy to promote and maintain overall general health. PMID- 15119283 TI - Starting anti-HIV therapy. AB - Deciding when to start anti-HIV therapy and what treatments to start with can leave many people feeling overwhelmed with choices. The discussion following, however, will demonstrate that the choices may be fewer and simpler than they appear at first. Charting your course of therapy options up front, outlining what therapies you will start with when you're ready and what you will switch to if that option doesn't work out, is the hallmark of long-term planning. You can then proceed to the next combination with confidence rather than being overwhelmed by the fact that your first choice didn't work out as you had expected. PMID- 15119284 TI - Opportunistic infection strategy. AB - The human immunodeficiency virus (HIV) infects immune cells, impairing their function and eventually destroying cells over time. This gradually weakens the immune system and the body loses the ability to fight disease. While HIV is the culprit, most people who die of AIDS do not die of HIV, per se, but from the numerous infections that the body can no longer control due to the collapse of the immune system. Relatively common infections, which may cause little or no harm in a healthy person, take the opportunity provided by weakened immune defenses to cause disease. This is why they are called opportunistic infections (OIs). PMID- 15119285 TI - What are STIs and what are the goals of STIs? AB - STIs (Structured Treatment Interruptions) involve going off anti-HIV therapy for periods of time in a structured and strategic fashion, typically guided by increased lab and health monitoring. In all, more than two dozen studies of STIs of varying types have been conducted since 1998. It is important to note that interpreting the results of STI research can be challenging. Some of the assumptions about HIV disease that led researchers to investigate treatment interruptions in the first place have yet to be proven conclusively. At least some of the research on STIs, however, has been promising and other research has made clear those areas where interrupting therapy is neither safe nor effective. PMID- 15119286 TI - Third line anti-HIV therapy options. AB - Third line therapy, sometimes called salvage or rescue therapy, is a term describing treatment regimens for people who have few or limited anti-HIV drug options. This includes people who have failed at least two previous anti-HIV regimens and/or people with evidence of HIV resistance to at least one drug in each of three major classes (NRTIs, NNRTIs and PIs, see the Drug ID chart on page 9 for more information on the drugs in each class). True salvage or deep salvage therapy is when a person has literally no viable treatment options. Treatment failure is a general term that encompasses a number of reasons that a regimen is deemed to be not working. The specific reasons for failure determines if or how the individual drugs in a regimen might be used again as part of a future combination. PMID- 15119287 TI - Immune strategy. AB - AIDS is a disease of primary immune dysfunction caused by HIV. To date, all of the approved and proven strategies for treating HIV disease focus on crippling the virus' ability to infect and/or destroy these cells. There are no approved treatments directed toward the immune deficits and dysfunctions caused by HIV. The good news is that when HIV replication is slowed through the use of anti-HIV drugs, the immune system begins to repair itself and there is evidence of a degree of immune restoration when HIV is controlled over time. PMID- 15119288 TI - Antiretroviral pipeline: new-drug reports from retroviruses conference. PMID- 15119289 TI - Micronutrient supplementation shows promise in placebo-controlled trial. AB - A supplement containing 33 vitamins, minerals, and antioxidants seemed to help persons with HIV. PMID- 15119290 TI - Nevirapine precautions published. AB - New instructions tell physicians who is most at risk for rare but serious side effects when starting this important drug. PMID- 15119291 TI - "Poppers," some other drugs, may increase HIV infection risk. AB - Use of amphetamines, hallucinogens, or inhaled nitrites was associated with a higher risk of HIV infection--an incidental finding in data from a recent HIV vaccine trial. PMID- 15119292 TI - New testing for very early HIV diagnosis. AB - Because persons newly infected with HIV may be especially infectious before the body has created antibodies to partially control the infection, there is a new public-health push to also look for the virus itself in routine HIV testing. North Carolina has used this approach statewide for over a year, and discovered the first indications of a new HIV epidemic among college students, especially African-American men. Results were reported at the recent Retroviruses conference. PMID- 15119293 TI - Improving AIDS conferences with online information. AB - Today, scientists and others arrive at major conferences without knowing whom they should meet and talk to outside of their own field. The whole medical research enterprise is damaged when researchers miss these connections. The key to improvement is to have the main data presentations online, allowing conferences to focus on exploration and discussion, instead of lectures that must rush through the new data. PMID- 15119294 TI - Buffalo hump: what the experts suggest. PMID- 15119295 TI - New nevirapine label underscores the risk for hepatotoxicity in key populations. PMID- 15119296 TI - Healthy eating. Just the two of us. Cooking for one or two. PMID- 15119297 TI - About alcohol. The heart-blood glucose connection. PMID- 15119298 TI - Unproven therapies. Evaluating new treatment options. PMID- 15119299 TI - A gut response. The next generation of type 2 drugs. PMID- 15119300 TI - One-pot meals. PMID- 15119301 TI - Defining success. Realistic goals are key to staying motivated about weight loss. PMID- 15119302 TI - Weight loss 101. Follow healthy eating advice or enjoy your meals? With these real-life strategies, you can do both! PMID- 15119303 TI - Rethinking Atkins. New research suggests that the famous low-carb diet may be safe--at least in the short term. PMID- 15119304 TI - Research profile. Seniors and risk. Too much glucose from the liver? Annette M. Chang, MD, MS. PMID- 15119305 TI - Support groups for people with HIV. PMID- 15119306 TI - Sex & sobriety: a support group study. PMID- 15119307 TI - Report from San Francisco: The 11th Conference on Retroviruses and Opportunistic Infections (CROI). The tenacity of NNRTI drug resistance. PMID- 15119308 TI - Report from San Francisco: The 11th Conference on Retroviruses and Opportunistic Infections (CROI). Antiretroviral therapy: the naive patient. PMID- 15119309 TI - Report from San Francisco: The 11th Conference on Retroviruses and Opportunistic Infections (CROI). The treatment of experienced patients and resistance mechanisms. PMID- 15119310 TI - Report from San Francisco: The 11th Conference on Retroviruses and Opportunistic Infections (CROI). Hepatitis: glimpses of optimism tempered by sobering mortality statistics. PMID- 15119311 TI - Report from San Francisco: The 11th Conference on Retroviruses and Opportunistic Infections (CROI). Complications of antiretroviral therapies and HIV. PMID- 15119312 TI - Report from San Francisco: The 11th Conference on Retroviruses and Opportunistic Infections (CROI). Lipodystrophy and metabolic complications. PMID- 15119313 TI - Early treatment of HIV. PMID- 15119314 TI - [New therapeutic strategies for Hodgkin lymphoma in cooperation of radiation oncology and medical oncology]. AB - BACKGROUND: Between 1984 and 2002 more than 9500 pts. were enrolled in the multicentric randomized trials of the German Hodgkin Study Group (GHSG) and are evaluable for response, survival, recurrences, and toxicities. Actually the GHSG evaluates the efficacy of risk-adapted therapy composed of polychemotherapy (CT) and radiotherapy (RT). An extensive RT quality assurance program has been practiced during the study generations and will be continued. PATIENTS AND METHODS: The 4th study generation (1998-2002) includes the following trials: In the HD10 trial (early stages) 4x ABVD are tested against 2x ABVD followed by 20 Gy Involved Field (IF)-RT vs. 30 Gy IF-RT (4 arms). In order to optimize CT regimen and IF-RT dose for pts. with intermediate stage, the HD11 trial compares 4x ABVD with 4x BEACOPP baseline followed by 20 Gy IF-RT vs. 30 Gy IF-RT in a 4 arm design. Concerning advanced stages (HD12), the BEACOPP regimen is to be optimized and the necessity of additive RT is tested. The standard arm (8x BEACOPP escalated) is compared with the toxicity reduced arm (4x BEACOPP escalated + 4x BEACOPP baseline) followed by 30 Gy RT on initial bulky disease and/or residual tumor vs. no RT (4 arms). RESULTS: Interim results (without arm comparisons) with a median follow-up of 18 months for HD10 and HD11 and 20 months for HD12 are as follows: Freedom from Treatment Failure (FFTF) at 18 months is in the HD10 trial (390 pts.) 96.4%, in the HD11 trial (480 pts.) 91.5% and in the HD12 trial (550 pts.) 90.2%.The overall survival (OS) at 18 months is in HD10 98.2%, in HD11 98.5%, in HD12 93.5%. In HD10, HD11 and HD12 respectively, 1.8%, 1.9% and 2.5% of pts. died and 1.0%, 2.5% and 2.2% suffered early progression. CONCLUSION/FURTHER STRATEGY: In order to reduce the relapse rate and toxicity and to improve the quality of life, the new HD13 trial for early stages (Fig. 1a) compares 2x ABVD, 2x ABV, 2x AVD und 2x AV, each followed by 30 Gy IF-RT. For the intermediate stages, the FFTF rate should be improved by intensifying the standard regimen. Therefore the new trial HD14 (Fig. 1b) compares 4x ABVD with 2x BEACOPP escalated + 2x ABVD, each followed by 30 Gy IF-RT. In the new trial for advanced stages HD15 (Fig. 1c),the FFTF/OS rates are to be maintained and the quality of life to be improved. 8x BEACOPP escalated, 6x BEACOPP escalated and 8x BEACOPP baseline with shortened 14-day cycle are compared. The use of PET to decide on additive RT will also be investigated. PMID- 15119315 TI - [Physiopathologic relationship between interstitial cystitis and rheumatic, autoimmune, and chronic inflammatory diseases]. AB - OBJECTIVES: To evaluate the current knowledge about interstitial cystitis pathophysiology and its relationship with rheumatic, autoimmune and chronic inflammatory diseases. METHODS: Literature search under "interstitial cystitis pathophysiology" either clinical or experimental trials and reports, in Medline, PubMed, Digital Urology Journal and Doctor's Guide, in addition to our own clinical research experience results. RESULTS: Both human and experimental trials show resemblances between interstitial cystitis and rheumatic, autoimmune, and chronic inflammatory diseases on clinical presentations, pathophysiology. Some interstitial cystitis patients show the bladder infiltrated with specific mononuclear cells, high incidence of circulating antinuclear antibodies, good response to anti-inflammatory and/or immunosuppressive therapies. Interstitial cystitis in association with rheumatic, autoimmune and chronic inflammatory diseases is very common. Many patients with systemic lupus erythematosus, Sjogren syndrome and fibromyalgia syndrome show antibodies against urothelium and/or muscle cells and/or other connective tissue components of urinary bladder. Systemic lupus erythematosus and Sjogren syndrome are the autoimmune diseases which bear strongest similarity with interstitial cystitis. Moreover, rheumatoid arthritis, chronic pelvic pain syndrome, irritable bowel syndrome, fibromyalgia, chronic fatigue syndrome, Evan's syndrome and atopic dermatitis share some pathogenic characteristics. CONCLUSIONS: Nowadays, interstitial cystitis pathophysiology is unknown. Based on clinical presentations, epidemiology, pathology and laboratory findings and treatment response, there is an important correlation among interstitial cystitis and rheumatic, autoimmune and chronic inflammatory diseases. These disorders may share some pathophysiologic mechanisms. Rigorous studies of pathophysiology of these group of diseases are needed to confirm consistently this approach for such conditions. PMID- 15119316 TI - [The concept of wound bed conditioning in chronic wounds]. PMID- 15119317 TI - Symposium. Foreword. [Joint Conference on Legal/Ethical Issues in the Progression of Dementia]. PMID- 15119318 TI - Ethics, ethnicity, and dementia: a "culture-fair" approach to bioethical advocacy in dementing illness. PMID- 15119319 TI - Decisionmaking at the end of life: patients with Alzheimer's or other dementias. PMID- 15119320 TI - Dementia research: ethics and policy for the twenty-first century. PMID- 15119321 TI - Thinking about life-sustaining treatment late in the life of a demented person. PMID- 15119322 TI - Predictive genetic testing for Alzheimer's disease in long-term care insurance. PMID- 15119323 TI - Dementia and Alzheimer's disease: an overview. PMID- 15119324 TI - Creating and supporting the proxy-decider: the lawyer-proxy relationship. PMID- 15119325 TI - Freedom fading: on dementia, best interests, and public safety. PMID- 15119326 TI - Radioecological software package: an interactive computational system to simulate the behaviour of radionuclides in semi-natural environments. AB - RSP (Radioecological Software Package) is an interactive support system that simulates the behaviour of radionuclides in semi-natural environments and the consequences on the population in terms of the external exposure. RSP consists of three modules: the first one, soil mobility, simulates the vertical transport of radionuclide in soil using the mathematical model RABES. The second module, soil to-plant transfer, simulates the radionuclide soil-to-plant transfer factor reported in the literature. Soil properties, vegetation types and environmental conditions are taken into consideration in the simulation process. In the third module, dose assessment, the dose-rate factor in air at a height of 1 m above ground can be calculated for sources distributed in a slab of finite thickness dn sources which are exponentially distributed with depth. The calculations are performed using DAGES Model, a Monte Carlo algorithm developed to simulated the photon transport for the soil/air configuration. A free copy of RSP can be downloaded from the website: http://imasl-apat.unsl.edu.ar. PMID- 15119327 TI - A brief history of the political work of genetics. AB - The biological sciences have long been used to define distinctions between people and to define inequalities as a natural consequence of essential biological traits. Today, geneticists draw distinctions on the basis of genetic predispositions. Their population-based methods can reinforce stereotypes about race and ethnic differences, providing concepts, validated by science, through which group differences can be interpreted as biologically ordained. Cases suggest how genetic variants can be used in social policies as individuals are differentially treated, not on the basis of their individual condition, but because of predispositions attributed to their group. PMID- 15119328 TI - Perspectives on research in American Indian communities. AB - This article discusses research-oriented responsibilities of the Zuni governor and tribal council to the Zuni people. To reduce potential negative effects and to enhance the lifestyle of the Zuni, these bodies screen and review research in an effort to ascertain compliance with tribal law, to be culturally respectful, and to determine what, if any, beneficial effects the research will have for the Zuni people. As a result, studies concerning high prevalence disease, such as diabetes, are given preference. These principles may apply to other American Indian and Alaskan native communities. PMID- 15119329 TI - Native American recommendations for genetic research to be culturally respectful. AB - This article describes genetic research issues and recommendations identified by inter-tribal Native American groups in meetings with tribal leaders from 1995 through 1999. PMID- 15119330 TI - A caution to Native American institutional review boards about scientism and censorship. AB - Native American Institutional Review Boards (IRBs) promote the health and welfare of tribes by reviewing protocols for research studies that focus on their tribes. The benefits of approved protocols should not be overstated lest good studies disappoint because they do not satisfy unachievable expectations. IRBs also should avoid the temptation to censor the outcomes of those studies. Science relies on candor and clarity about results and methods to move forward. PMID- 15119331 TI - An analysis of research guidelines on the collection and use of human biological materials from American Indian and Alaskan Native communities. AB - American Indian and Alaskan Native communities have expressed concern about the use of human biological materials in research. These concerns have prompted research sponsors and professional organizations to develop guidelines for investigators working with these communities. This paper reviews research guidelines and presents recommendations that reflect "best practices" for working with North American indigenous communities in the collection, storage, and distribution of human biological materials for research. These recommendations strike a reasonable balance between three imperatives in research: (1) minimizing harm, (2) treating sample contributors with respect, and (3) promoting intellectual freedom to pursue a range of research questions. The recommendations can be used in designing appropriate methods of collecting and using human biological materials from members of American Indian and Alaskan Native communities and will likely be applicable to other historically disadvantaged communities as well. PMID- 15119332 TI - A critical appraisal of protections for aboriginal communities in biomedical research. AB - As scientists target communities for research into the etiology, especially the genetic determinants of common diseases, there have been calls for the protection of communities. This paper identifies the distinct characteristics of aboriginal communities and their implications for research in these communities. It also contends that the framework in the Belmont Report is inadequate in this context and suggests a fourth principle of respect for communities. To explore how such a principle might be specified and operationalized, it reviews existing guidelines for protecting aboriginal communities and points out problems with these guidelines and areas for further work. PMID- 15119333 TI - Genetic research and communal narratives. AB - The use of DNA evidence to prove Thomas Jefferson's paternity of Sally Hemmings' children is a powerful example of how genetic research can have an impact upon the communal narratives of families and nations. PMID- 15119334 TI - [Winter viruses]. PMID- 15119335 TI - [Acute infections in the child]. PMID- 15119336 TI - Institutional review boards and social science research. PMID- 15119337 TI - New guidelines for protection of subjects who are prisoners. PMID- 15119338 TI - New waiver of regulations on research with prisoner-subjects. PMID- 15119339 TI - Cloning human cells for research called ethical. PMID- 15119340 TI - Military personnel may have been human subjects without their informed consent. PMID- 15119341 TI - Reducing the burden on many institutional review boards. PMID- 15119342 TI - Institutional review boards (IRBs) failed to use steps to protect vulnerable research subjects. PMID- 15119343 TI - Shaping the future depends on strengthening health systems. PMID- 15119344 TI - Substandard and counterfeit medicines. PMID- 15119345 TI - Adjuvant therapy for early staged (epithelial) cancer. PMID- 15119346 TI - Community engagement. The HSJ panel discussion. PMID- 15119347 TI - [Craniocerebral trauma]. PMID- 15119348 TI - Utilization of the relative complexity measure to construct a phylogenetic tree for fungi. AB - The relative complexity measure (RCM) is a new approach to evaluate relatedness of DNA sequences which eliminates the requirement to align sequences prior to analysis, a step required with standard reference methods. The value of the RCM approach to generate distance matrices for use in phylogenetic analysis of organisms has not been determined. This study compared RCM with the algorithmic and tree searching reference methods for phylogenetic analysis using fungal sequences. Sequences of the cytochrome b gene and the 18S rDNA gene were obtained from the GenBank database to determine feasibility of this method for phylogenetic relatedness. The RCM approach was also used to construct a phylogenetic tree using internal transcribed spacer (ITS) sequences from 23 medically relevant fungal species. The robustness of the RCM and reference approaches was determined by comparing the topology of seven medically relevant fungi within the phylogenetic trees generated after progressive removal of 10, 20, 30, 40 and 50% of the nucleotide bases from either the 5' or 3' end of the three genomic target sequences. The results demonstrated that the RCM method was equivalent to the reference methods for construction of phylogenetic trees from cytochrome b and 18S rDNA gene sequences. The phylogenetic tree constructed using the ITS sequence generated no contradictory topology. The RCM generated trees retained the appropriate topology after removal of up to 50% of the cytochrome b sequence, 40% of the ITS sequence, and 30% of the 18S gene target sequence. Comparatively, the reference methods failed to maintain topology after only a 10% sequence deletion for each genomic target. The results showed the RCM to be a reliable and robust computational approach for use in the construction of fungal phylogenetic trees without the requirement for prior sequence alignment. PMID- 15119349 TI - Pleomorphic conidiation in Claviceps. AB - Types of asexual sporulation in 17 Claviceps species and the closely related Corallocytostroma ornicopreoides were revised in relation to the phylogeny of clavicipitaceous fungi. We observed: (1) enteroblastic conidiation from branched phialidic conidiophores typical of the genus (anamorph Sphacelia) in all species including Corallocytostroma; (2) widespread and often sequential formation of terminal holoblastic secondary conidia on tapering hyphae arising from sphacelial macroconidia: and (3) in addition to sphacelial conidiation, sympodial holoblastic conidiation of the Ephelis-type in cultures of C. zizaniae and in both the culture and sphacelial tissue of C. citrina. Secondary conidiation was not found in C. purpurea, C. citrina and C. sorghicola. During sphacelial fructification, most species produced macroconidia and microconidia. Only macroconidia formed in planta underwent secondary conidiation whereas microconidia did not germinate at all. In C. phalaridis, the formation of holoblastic 2-3 celled appendaged conidia was observed, similar to that of Aciculosporium and Neoclaviceps. In dendrograms based on ITS1-5.8S rDNA-ITS2 sequences, genera and species with appendaged conidia grouped on a highly supported clade with ancestral Corallocytostroma. The clade was placed inside a group of tropical species of Claviceps, without any relationship to Balansiae. PMID- 15119350 TI - Sequence polymorphism and molecular characterization of laccase genes of the conifer pathogen Heterobasidion annosum. AB - The oxidizing enzyme laccase produced by many fungi is generally considered to be active in the biodegradation of lignin, a major plant cell wall component highly resistant to microbial attack. The enzyme is secreted at high levels by the P type of the highly aggressive pathogen Heterobasidion annosum, but at much lower levels by the S-type which correlated with their varying wood decay capability. To investigate the evolutionary relationship between laccase genes of the different H. annosum types from several geographical regions we have compared the nucleotide sequence of the laccase gene from 32 different isolates of the fungus together with two other Asian isolates (H. araucariae, H. insulare). In addition to nucleotide sequence assessment, we have also cloned, characterized and analysed the partial sequences of the laccase gene from the homokaryotic (FSE-7: S-type; Sa16-4: P-type) and heterokaryotic (Faf-8: F-type) strains of H. annosum. Using degenerate primers, two distinct laccase gene fragments of 1.64 and 2 kb were amplified from the genomic DNA of this fungus. DNA sequence analyses showed that the 1.64 kb laccase fragment in all three H. annosum types shared significant homology (86-96%). But comparative analyses of the 1.64 and 2 kb laccase gene fragment revealed only 48% nucleotide sequence similarity. Using the cDNA sequence information, exon regions were predicted and this revealed that about nine small introns interrupted the genomic DNA. Southern hybridization analysis indicated a single copy of the gene in the homokaryotic S-type (FSE-7) examined but presence of double bands in the homokaryotic and heterokaryotic P type strains of the fungus suggest the existence of two laccase genes. Northern analyses revealed that the gene is constitutively expressed but appear to be enhanced several fold with the addition of ferulic acid or oxalic acid. Alignments of the nucleotide sequences and phylogenetic analyses are presented allowing estimations of evolutionary relationships to be made. These comparisons indicate that laccase gene of P-type is distinct from other Heterobasidion sequences, including the outgroups H. araucariae and H. insulare, while the relationship between the S- and F-groups could not be resolved. Comparative phylogenetic analyses using predicted amino acid sequence also showed strong similarity to the laccases from other basidiomycetes (Pleurotus ostreatus, Phlebia radiata and Trametes versicolor) but least similar to laccases from ascomycete fungi. In addition, the results of McDonald-Kreitman test for possible evidence of selection based on analyses of two exon regions of the H. annosum laccase gene are presented and discussed. PMID- 15119351 TI - Double-stranded RNA transmission through basidiospores of Heterobasidion annosum. AB - A search for double-stranded RNA (dsRNA) was conducted among 106 isolates of the pathogenic basidiomycete Heterobasidion annosum. Of these isolates, 47 were tissue isolates from fruit bodies and 59 were isolated from decayed wood. Nucleic acids were extracted from freeze-dried mycelia and dsRNA was separated by cellulose CF-11 chromatography and confirmed by digestions with specific nucleases. dsRNA was present in 19 and 14% of the tissue and wood isolates, respectively. From five of the fruit bodies containing dsRNA basidiospores were investigated and 10-84% of the germinated basidiospores contained dsRNA. On high nutrient media, the germination frequency of basidiospores was reduced by presence of dsRNA in the fruit body (P < 0.05); germination frequencies were 34 and 78% for spores from fruit bodies with and without dsRNA, respectively. The same trend was present also on low nutrient media, although not statistically significant; germination was 3 and 10% for spores from infected and dsRNA free fruit bodies, respectively. Transmission of dsRNA in H. annosum from mycelia into basidiospores together with the lowered germination frequency are likely to play a significant role in the life cycle of the fungus. The relative importance of different transmission routes for dsRNA in H. annosum is discussed. PMID- 15119352 TI - Fusaproliferin, beauvericin and fumonisin production by different mating populations among the Gibberella fujikuroi complex isolated from maize. AB - The production of fumonisins, fusaproliferin and beauvericin by Gibberella fujikuroi different mating populations isolated from maize in Argentina was evaluated. From 203 strains of Fusarium verticillioides (G. fujikuroi mating population A), 193 were fumonisin producers. Among members of mating population A, female fertile strains produced 20% more toxin than female sterile ones. Among 78 Fusarium proliferatum strains (G. fujikuroi mating population D) 65 produced fumonisins. The percentage of strains that were high, intermediate and low level toxin producers varied according to the species evaluated and the area from which the strains were isolated. Fusarium subglutinans (G. fujikuroi mating population E) strains produced low levels or were no fumonisin producers. Strains from both G. fujikuroi mating populations D and E were able to produce fusaproliferin and beauvericin. Among the members of F. subglutinans (G. fujikuroi mating population E) the fusaproliferin production was more constant. Co-production of fumonisin, fusaproliferin and beauvericin among the strains belonging to G. fujikuroi D and E was also observed. The co-production of fumonisin, beauvericin and fusaproliferin in maize need to be considered, since from the toxicological point of view interactions between these toxins could occur. The toxigenic ability of the strains evaluated prompt us that is necessary to determine the natural occurrence of fusaproliferin and beauvericin in Argentinean maize. PMID- 15119353 TI - The biocontrol fungus Pochonia chlamydosporia shows nematode host preference at the infraspecific level. AB - A RAPD-PCR assay was developed and used to test for competitive variability in growth of the nematode biological control fungus Pochonia chlamydosporia. Saprophytic competence in soil with or without tomato plants was examined in three isolates of the fungus: RES 280 (J), originally isolated from potato cyst nematode (PCN) cysts; RES 200 (I) and RES 279 (S), both originally isolated from root knot nematode (RKN) eggs. Viable counts taken at 70 d indicated that I was the best saprophyte followed by S, with J the poorest. RAPD-PCR analysis of colonies from mixed treatments revealed that there was a cumulative effect of adding isolates to the system. This suggested that the isolates did not interact and that they may occupy separate niches in soil and the rhizosphere. To investigate parasitic ability, soils were seeded with two isolates of the fungus: J and S, singly or in combination. Tomato or potato plants were grown in these soils: free of nematodes, or inoculated with PCN or RKN, and incubated for 77 d. The abundance of the PCN isolate J in PCN cysts was significantly greater than that of the RKN isolate S but in RKN egg masses, S was significantly more abundant than J. RAPD-PCR analysis of colonies from mixed treatments confirmed that J was more abundant than S in PCN cysts whereas the converse was observed on RKN egg masses. This substantiates the phenomenon of nematode host preference at the infraspecific level of P. chlamydosporia and highlights its relevance for biological control of plant parasitic nematodes. PMID- 15119354 TI - Histological studies on the mycoparasitism of Cladosporium tenuissimum on urediniospores of Uromyces appendiculatus. AB - Interactions between the mycoparasite Cladosporium tenuissimum and the bean rust Uromyces appendiculatus were studied through light and electron microscopy in vitro at the host-parasite interface. Urediniospore germination decreased on contact with ungerminated C. tenuissimum conidia, possibly due to antibiosis mechanisms. C. tenuissimum grew towards the bean rust spores and coiled around their germ tubes. Penetration of the urediniospores occurred either enzymatically and/or mechanically, through appressorium or infection cushion structures, from which a thin penetrating hypha was generated. Enzyme production by the mycoparasite was suggested by the loosening of the matricial components of the spore wall, which sometimes left chitin fibrils visible. Mycoparasite hyphae grew within the host spore, emptied its content, and emerged profusely forming conidiophores and conidia. C. tenuissimum was able to grow on media containing laminarin, suggesting the ability of producing glucanases, but not when chitin was used as the sole carbon source. Conidia that had been grown on a sugar-rich medium, filtered, and extracted with organic solvents, were found to contain cladosporol and related compounds. Complete control of the bean rust disease was achieved by application of C. tenuissimum culture filtrates but not by conidial suspensions. This is the first report of parasitism by C. tenuissimum on U. appendiculatus. These investigations provide additional observations on a genus besides Melampsora and Cronartium from which this fungus has been isolated and tested to date. The possible role of environmental factors for the exploitation of this organism as a biocontrol agent is also mentioned. PMID- 15119355 TI - Effects of available water on growth and competition of southern pine beetle associated fungi. AB - Competitive, interactions among bark beetle associated fungi are potentially influenced by abiotic factors. Water potential, in particular, undergoes marked changes over the course of beetle colonization of tree hosts. To investigate the impact of water potential on competition among three southern pine beetle associated fungi, Ophiostoma minus, Entomocorticium sp. A and Ceratocystiopsis ranaculosus, we utilized artificial media with water potentials of 0, -5, -10, and -20 MPa. Growth of all three fungi, when grown alone, decreased on media with lower water potentials. Growth rates of all three fungi were likewise reduced in competition experiments. At -5 to -10 MPa, C. ranaculosus (a fungus with beneficial effects toward southern pine beetle) was nearly equal in competitive ability to O. minus (a fungus with antagonistic effects towards southern pine beetle). This was not true on control media, nor at other water potentials tested. The range of water potentials used in our assays was similar to the range of water potentials we measured in loblolly pines within a southern pine beetle infestation. This study indicates that water potential may alter the outcome of competitive interactions among bark beetle-associated fungi in ways that favour bark beetle success. PMID- 15119356 TI - Growth and interspecific interactions of the rare oak polypore Piptoporus quercinus. AB - Growth and interactions of Piptoporus quercinus were studied in artificial culture to give insights into the ecological characteristics of a species which is both rare and widespread. P. quercinus grew slowly under all conditions, the maximum ranging between 1.9 and 3.15 mm d(-1), depending on isolate, at 25 degrees C pH 3.75. Upper limits for growth were 25-30 degrees or above 30 degrees depending on isolate, and pH 6.6. Lower limits for growth were 5-10 degrees, between -1.75 and -2.5 MPa; there was still growth at pH 1.81. Preincubating plates at 5 for 77 d and then incubating at 20 degrees resulted in a significantly greater extension rate (2.89 mm d(-1)) than when incubated continuously at 20 degrees (1.70 mm d(-1)) for one isolate though not for another isolate. P. quercinus survived exposure to low temperature (-18 degrees for 7 d), showing renewed growth, albeit limited, after a lag of 37 d. It was a poor combatant, being unable to replace any of the eight fungi (with a range of ecological roles) against which it was paired, and being replaced or partially replaced by most, on 2% MA at pH 5.49, at pH 3.75 and at -1.25 MPa. It did, however, inhibit extension rate of some other heart rot fungi. Results are discussed in an ecological context. PMID- 15119357 TI - Phylogenetic analysis of the downy mildew pathogen of oilseed poppy in Tasmania, and its detection by PCR. AB - Downy mildew of oilseed poppy (Papaver somniferum) has become a serious disease issue for the Tasmanian poppy industry since its first record in 1996. Previous reports have reported the pathogen as Peronospora arborescens, which is differentiated from the related species P. cristata, also known to infect Papaver spp., by conidium dimensions alone. This study investigated the taxonomic status of the downy mildew pathogen, using both morphological characters and molecular analysis of the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA). The inherent variability of conidium dimensions made differentiation of species difficult. Sequence homology and phylogenetic analyses of the ITS region showed the pathogen to be more closely related to P. cristata than P. arborescens. It is therefore proposed that downy mildew of oilseed poppy in Tasmania be reattributed to the pathogen P. cristata. In addition to this work, PCR primers have been developed for the specific detection of the downy mildew pathogen in Tasmania. PMID- 15119358 TI - Description and affinities of a new sequestrate fungus, Barcheria willisiana gen. et sp. nov. (Agaricales) from Australia. AB - A new sequestrate fungus, Barcheria willisiana gen. et sp. nov., is described and its affinities evaluated using nLSU rDNA sequence data. This unusual fungus has several characters that are reminiscent of species of Agaricus and Lepiota, but with a very reduced basidiome form. The nLSU rDNA of four Australian taxa, Barcheria willisiana, Agaricus xanthodermus, Leucoagaricus naucinus, and Lepiota discolorata, was sequenced for this study. Parsimony analysis of the sequences placed Barcheria within an Agaricus clade. PMID- 15119359 TI - Gram-negative bacillary meningitis after cranial surgery or trauma in adults. AB - In order to assess the clinical features, aetiology, treatment and outcome of post-neurosurgical and post-traumatic Gram-negative bacillary meningitis (GNBM) we performed a retrospective review of all adult patients admitted to the Department of Neurosurgery who had Gram-negative bacilli cultured from cerebrospinal fluid (CSF) following a neurosurgical procedure or traumatic head/spinal injury. During the 12 y of the review 33 patients had CSF isolates of Gram-negative bacilli that were thought to be significant. The median patient age was 47 y (range 22-77 y) and 21 (64%) were male. Klebsiella pneumoniae, Enterobacter cloacae and Escherichia coli were the most common isolates. Minimal inhibitory concentrations (MIC) measured for half the patients' isolates resulted in 5 regimen changes, including 2 patients with E. cloacae meningitis in whom cephalosporin susceptibility decreased during cephalosporin treatment. Our recommended initial treatment was intravenous ceftriaxone and amikacin, subsequently tailored by susceptibility results; approximately half the patients remained on the antibiotics they started and half were changed to an alternate regimen, most often a carbapenem. Five patients (15%) died, 1 dying after cure of his GNBM. There were no failures in those who received more than 12 d of appropriate treatment: treatment for at least 14 d after the last positive CSF culture guaranteed cure. Initial ceftriaxone and amikacin subsequently changing to susceptibility driven alternatives, often a carbapenem, resulted in cure of 85% of our patients with GNBM. PMID- 15119360 TI - Investigation of Burkholderia cepacia nosocomial outbreak with high fatality in patients suffering from diseases other than cystic fibrosis. AB - Over a 1-y period, 26 inpatients at the Jordan University Hospital in Amman were detected with bacteraemia (23 cases) or respiratory tract colonized with B. cepacia (3 cases). A combination of genetic identification and molecular typing has proved that all cases were caused by a single epidemic strain of B. cepacia genomovar IIIa. Nosocomial infections could be documented in 21/26 (81%) patients, mostly with severe underlying or malignant diseases other than cystic fibrosis, but the source of infection was undetected. The overall mortality related to infection with B. cepacia was 42%. All B. cepacia isolates were resistant to ampicillin, amikacin, carbenicillin and gentamicin; and mostly susceptible to piperacillin, chloramphenicol, cotri-moxazole, tetracycline, ceftazidime, and tazocin (62-88%). This study demonstrates the nosocomial and high fatality of B. cepacia genomovar IIIa in Jordanian patients suffering from diseases other than cystic fibrosis. PMID- 15119361 TI - Primary herpes simplex virus type 1 gingivostomatitis during the second and third trimester of pregnancy: foetal and pregnancy outcome. AB - Primary herpetic gingivostomatitis during pregnancy is a relatively rare phenomenon with no clear management guidelines. We describe 4 cases of primary herpetic gingivostomatitis during pregnancy and review the literature. The diagnosis in all cases was based on clinical manifestations and serology and culture findings. Two of the women received intravenous rehydration. Acyclovir, offered in 2 cases, was refused. Delivery was normal in all 4 cases, with good neonatal outcome, similar to findings in the literature (6 cases). Herpetic gingivostomatitis in the second and third trimester of pregnancy does not appear to be associated with adverse foetal effects. More data are needed to define the risk of this infection during pregnancy. PMID- 15119362 TI - Serum ALT levels as a surrogate marker for serum HBV DNA levels in HBeAg-negative pregnant women. AB - In Stockholm, Sweden, the majority of pregnant women positive for hepatitis B surface antigen (HBsAg) are hepatitis Be antigen (HBeAg) negative. Newborns to HBeAg positive mothers receive vaccination and hepatitis B immunoglobulin (HBIg). Newborns to HBeAg negative mothers receive vaccine and HBIg only if the mothers have elevated ALT levels. The aim of this study was to retrospectively evaluate ALT levels as a surrogate marker for HBV DNA levels in HBeAg negative carrier mothers. Altogether 8947 pregnant women were screened for HBV markers from 1999 to 2001 at the Virology Department, Karolinska Hospital. Among mothers screened 192 tested positive for HBsAg (2.2%). 13 of these samples could not be retrieved. Of the remaining 179 sera, 8 (4%) tested positive for HBeAg and 171 (95.5%) were HBeAg negative. Among the HBeAg negative mothers, 9 had HBV DNA levels > 10(5) copies/ml, and of these 7 had normal ALT levels indicating low sensitivity of an elevated ALT level as a surrogate marker for high HBV DNA level. Furthermore, no correlation was found between ALT and HBV DNA levels. Hence, it is concluded that the use of ALT as a surrogate marker for high viral replication in HBeAg negative mothers could be questioned. PMID- 15119363 TI - Body composition changes in 308 Norwegian HIV-positive patients. AB - This is the first study where HIV-associated body composition changes are described in a Scandinavian cohort. All HIV-positive patients living in Oslo who attended our outpatient clinic (n = 407) were invited to participate. 308 patients (78%) were included. Lipodystrophy (LD) prevalence was 37.3% in patients on antiretroviral therapy (ART+) compared to 10.9% in patients without ART (p < 0.001). Prominent veins and combined fat atrophy/accumulation were exclusively found in the ART+ group. Determinants of prominent veins were skin fold thickness, duration of nucleoside reverse transcriptase inhibitor treatment, duration of protease inhibitor treatment and current use of stavudine. When patients with and without LD were compared, breast circumference was 10.6 cm larger in LD+ women than in LD- women (p = 0.003). Chest pain was reported in 26.5% of LD+ compared to 3.9% (p < 0.001) of LD- patients. This may be associated with an increased level of creatin kinase in LD+ compared to LD- patients (161 +/ 179 U/I vs 102 +/- 68, p = 0.004). Eight years after HIV diagnosis 59.1% of the patients with LD had a regular job and 59.4% reported no or small problems with ART. PMID- 15119364 TI - The use of CRP tests in patients with respiratory tract infections in primary care in Sweden can be questioned. AB - A diagnosis-antibiotic prescribing study was performed in 5 counties in Sweden during 1 week in November in 2000 and 2002 respectively. As a part of the study, the use and results of C-reactive protein (CRP) tests in relation to duration of symptoms and antibiotic prescribing in 6778 patients assigned a diagnosis of respiratory tract infections were analysed. In almost half (42%) of the patients, a CRP test was performed. The majority of CRP tests (69%) were performed in patients assigned diagnosis upper respiratory tract infection, where the test is not recommended. Overall, there was a minor decrease in antibiotic prescribing when CRP was used (41%), in comparison to 44% of the patients where no CRP was performed (p < 0.01). Patients assigned diagnoses implying a bacterial aetiology were prescribed antibiotics irrespective of result of CRP or length of symptoms before consultation. For patients assigned viral diagnoses, antibiotic prescribing increased with increasing duration of symptoms and increasing value of CRP. The use of CRP decreased antibiotic prescribing in patients assigned to viral diagnoses and with longstanding symptoms (p < 0.001). However, 59% of the patients assigned viral diagnoses with CRP > or = 25 received antibiotics, which seems to indicate a misinterpretation of CRP and a non-optimal use of antibiotics. PMID- 15119365 TI - Diagnostic value of bronchoalveolar lavage in community-acquired pneumonia in a routine setting: a study on patients treated in a Finnish university hospital. AB - Only a few previous studies have focused on the use or bronchoalveolar lavage (BAL) in patients with community-acquired pneumonia (CAP). Our aim was to evaluate the diagnostic value of BAL in CAP in a routine clinical setting. 71 disease episodes were retrospectively analysed. The patients had undergone BAL for serious or slowly responding pneumonia. All procedures were performed during antimicrobial treatment of the patient. BAL fluid was cultivated for bacteria, fungi, and viruses. In 68 episodes, 1 or several specific polymerase chain reaction tests were performed. Only 1 (1.3%) quantitative bacterial culture was considered diagnostic for CAP, and indicated a change of antimicrobial treatment. The diagnostic yield increased to 9.8% when other methods were used. A respiratory virus was the only aetiology in 3 (6.0%) patients. In slowly responding pneumonia, also hospital-acquired pathogens and malignancies were identified, resulting in a total diagnostic yield of 20.0%. Thus, even when a large array of diagnostic assays was applied, the value of BAL in pretreated patients with CAP was very small, and its therapeutic implications minimal. In a subgroup of slowly responding pneumonia, the procedure was of some usefulness even after commencement of antimicrobial treatment. PMID- 15119366 TI - Diarrhoea in elderly people: aetiology, and clinical characteristics. AB - We examined the characteristics of elderly people attending the Dhaka Hospital of ICDDR,B. The hospital has a diarrhoeal disease surveillance system that enrols a 2% systematic sample of all patients visiting the hospital. We reviewed data of all patients enrolled into the surveillance system (n = 13,782) over the period 1996-2001 to identify patients aged 60 y and above for inclusion into the current study (4% of all surveillance patients; n = 478). V. cholerae O1 was the most common enteric pathogen isolated from faecal culture of the patients (20%), followed by ETEC (13%), Shigella (11%), V. cholerae O139 (10%), Campylobacter jejuni (5%), Salmonella (3%), EPEC (2%), rotavirus (4%), and E. histolytica (2%). The isolation rate of V. cholerae O139 and Shigella was higher among the elderly compared to adults (15-59 y of age, 10% vs 6%, and 11% vs 7% respectively; p < 0.05 for both comparisons). Compared to 15-59-y-olds, a significantly higher proportion of the elderly had visible blood in stools (8% vs 5%), required short stay ward admission (86% vs 82%) or referral (1% vs < 1%) to a health facility. Early initiation of oral or i.v. rehydration therapy, prompt referral, and immediate clinical diagnosis for assessment of the need for antibiotic therapy might be beneficial for the elderly. PMID- 15119367 TI - Cold bone defect on granulocytes labelled with technetium-99m-HMPAO scintigraphy: significance and usefulness for diagnosis and follow-up of osteoarticular infections. AB - We wished to determine the frequency and significance of cold bone defect on granulocytes labelled with technetium-99-m-hexamethylpropyleneamine oxime (99mTc HMPAO-PMN) in non-spinal bone infection. Cold bone defect was investigated as part of a retrospective review during a 2-y period. Patients who had possible osteoarticular infection underwent bone scintigraphy combined with 99mTc-HMPAO PMN for diagnosis and follow-up. Osteomyelitis was confirmed by isolation of the responsible pathogen. Among 210 patients who had possible infection, 17 (8%) demonstrated a cold bone defect. The site of cold bone defect was for all patients the hip. All 17 patients had proven bacterial orthopaedic hardware related infection. The single causative micro-organism was staphylococcus. Whatever the outcome, cold bone defect was constant regardless of follow-up equal to or longer than 18 months. These data suggest that this uncommon scintigraphic pattern is an indication of an infectious process similar to increased uptake. PMID- 15119368 TI - Excessive use of rapid tests in respiratory tract infections in Swedish primary health care. AB - A 1-y retrospective study of problem oriented electronic patient records, for encounters concerning respiratory tract infection, was performed. The aim was to analyse the management of respiratory tract infections in primary health care in terms of diagnostic coding, tests and antibiotic treatment using data from electronic patient records. 12 primary health care centres with a registered population of 102,050 residents in 3 counties in southeast Sweden participated. Data were retrieved electronically from records of patient encounters concerning respiratory tract infections. The data were: patient age and gender, date of contact, diagnostic code, CRP and GABHS tests and results, as well as antibiotic prescriptions. In a total of 19,965 encounters, the most frequent diagnoses were common cold (40%), acute tonsillitis (18%), and acute bronchitis (15%). A total of 4445 GABHS tests (in 22% of encounters) and 6141 CRP tests (31%) were performed, and both tests were done in 1910 encounters (10%). A total of 7934 antibiotic prescriptions were registered. The proportion of patients tested and prescribed an antibiotic varied greatly between centres. We found an excessive, and much varying, use of rapid tests in encounters for respiratory tract infections. Data retrieval from electronic patient record systems was a feasible method to study the use of laboratory tests in relation to pharmacological treatment. PMID- 15119369 TI - Prevalence of non-O157 Escherichia coli strains among shiga-like toxin-producing (SLTEC) isolates in the region of Lower Silesia, Poland. AB - During 5 y (1997-2002) in the region of Lower Silesia, Poland, 55 shiga-like toxin-producing strains were isolated from children with diarrhoea, none of whom developed haemolytic uremic syndrome. Shiga-like toxin production was detected on Vero cells and confirmed by the detection of slt1 and slt2 gene sequences. Most of isolates examined possessed slt2 gene alone (41.8%), or in combination with slt1 gene (38.2%). None of the shiga-like toxin-producing strains was of serotype O157:H7. It appears that non-O157 shiga-like toxin-producing E. coil strains are prevalent as the causal agent of severe diarrhoea in the region. PMID- 15119370 TI - Successful treatment of disseminated cerebritis complicating methicillin resistant Staphylococcus aureus Endocarditis unresponsive to vancomycin therapy with linezolid. AB - A unique case of community acquired methicillin resistant Staphylococcus aureus (MRSA) sepsis, with endocardial and cerebral metastatic seeding, caused by a strain representative of the Italian clone, is described. The patient was a 47-y old man without apparent risk factors for endocarditis and for MRSA infection who developed coma with multiple cerebritis lesions under vancomycin plus amikacin therapy. He was eventually cured with the addition of linezolid to the initial antimicrobial regimen. This observation seems to confirm previous reports of the efficacy of linezolid for the treatment of central nervous system infections caused by multidrug resistant Gram-positive bacteria. To our knowledge, this is the first report of MRSA disseminated cerebritis, a nearly always fatal disease, cured with this oxazolidinone drug. The increase in community acquired MRSA may have some impact on empirical treatment of serious infections caused by this organism. PMID- 15119371 TI - Two case reports of symptomatic visceral leishmaniasis in AIDS patients concomitant with immune reconstitution due to antiretroviral therapy. AB - We report the first 2 cases of visceral leishmaniasis in AIDS patients, which ocurred a few d after initiating antiretroviral therapy. The report raises the question whether a rapid immune reconstitution may convert a latent visceral leishmaniasis into a symptomatic one. PMID- 15119372 TI - Submandibular lymph node enlargement due to cysticercosis infestation. AB - Lymph node enlargement due to cysticercus infestation has not been reported in human subjects. A 7-y-old girl presented with seizures and a right submandibular lymph node enlargement. Investigations showed inflammatory granulomas in the brain. Fine needle aspiration cytology from submandibular lymph node and enzyme linked immunosorbent assay of serum and cerebrospinal fluid showed the presence of cysticercosis infestation. The case underlines the potential of fine needle aspiration cytology for establishing diagnosis in patients having enlarged lymph nodes. PMID- 15119373 TI - Unusual pulmonary Enterocytozoon bieneusi microsporidiosis in an AIDS patient: case report and review. AB - Enterocytozoon bieneusi is an agent of intestinal microsporidiosis leading to chronic diarrhoea in AIDS patients. Pulmonary involvement may occur but remains rare with only 4 cases reported in the literature. We report here the fifth case of pulmonary localization of E. bieneusi in a severe immunocompromized HIV infected patient with intestinal and pulmonary symptoms. PMID- 15119374 TI - Co-occurrence of Epstein-Barr virus infection and ascites in sarcoidosis. AB - We present a young woman with a rare association of Epstein-Barr virus (EBV) infection, sarcoidosis and massive ascites. The temporal relationship between the EBV infection and ascites strongly suggests a pathophysiological relationship, and other causes of ascites were not identified. The prognosis of ascites is excellent in acute EBV infection. PMID- 15119375 TI - Thrombotic microangiopathy and hypothermia in an HIV-positive patient: importance of cytomegalovirus infection. AB - Cytomegalovirus (CMV) infection is a well-known frequent complication in HIV infected patients. We report a case of thrombotic microangiopathy and severe hypothermia in a 21-y-old woman positive for HIV infection. CMV infection was diagnosed by PCR. The symptoms completely resolved after ganciclovir therapy which supports the role of CMV as a causative agent. PMID- 15119376 TI - Group A streptococcal brain abscess. PMID- 15119377 TI - Lesions of the rat nucleus basalis magnocellularis disrupt appetitive-to-aversive transfer learning. AB - Rats with selective lesions of the nucleus basalis magnocellularis (NBM) and sham lesion control animals were tested in an operant appetitive-to-aversive transfer task. We hypothesized that NBM lesions would not affect performance in the appetitive phase, but that performance would be impaired during subsequent transfer to the aversive phase of the task. Additional groups of NBM lesion and control rats were tested in the avoidance condition only, where we hypothesized that NBM lesions would not disrupt performance. These hypotheses were based on the argument that the NBM is not necessary for simple association learning that does not tax attention. Both the appetitive phase of the transfer task and the avoidance only task depend only on simple associative learning and are argued not to tax attention. Consequently, performance in these tasks was predicted to be spared following NBM lesions. Complex, attention-demanding associative learning, however, is argued to depend on the NBM. Performance in the aversive phase of the transfer task is both attentionally demanding and associatively more complex than in either the appetitive or aversive tasks alone; thus, avoidance performance in the NBM lesion group was predicted to be impaired following transfer from prior appetitive conditioning. Results supported our hypotheses, with the NBM lesion group acquiring the appetitive response normally, but showing impaired performance following transfer to the aversive conditioning phase of the transfer task. Impairments were not attributable to disrupted avoidance learning per se, as avoidance behavior was normal in the NBM lesion group tested in the avoidance condition only. PMID- 15119378 TI - Neural substrates of olfactory discrimination learning with auditory secondary reinforcement. I. Contributions of the basolateral amygdaloid complex and orbitofrontal cortex. AB - The basolateral amygdaloid complex (BLA) and orbitofrontal cortex (OFC) share extensive reciprocal connections, and interactions between these regions likely contribute to both mnemonic and affective processes. The present study examined the potential differential contributions of the BLA and OFC to performance of an olfactory discrimination task that incorporates auditory conditioned reinforcement and to expression of immediate post-shock freezing behavior. Damage to the BLA had little effect on performance of the conditioned reinforcement task but abolished immediate post-shock freezing behavior. In contrast, damage to OFC resulted in both a mild but significant performance decrement in the conditioned reinforcement task and a significant attenuation of immediate post-shock freezing behavior. These findings suggest that immediate post-shock freezing behavior is likely critically dependent upon interactions between the BLA and OFC. However, although mnemonic processes underlying accurate performance of the conditioned reinforcement task might be supported by OFC in part, such processes are independent of either the BLA or interactions between these two regions. PMID- 15119379 TI - Idiodynamic profiles of cardiovascular activity: a P-technique approach. AB - A study was conducted to expand the conventional view of cardiovascular (CV) reactivity by using the idiodynamic paradigm for investigation of individuals. Patterns of autonomic CV regulation were assessed in six subjects across diverse laboratory tasks on three separate occasions. Individual CV profiles were derived from these data with P-technique factor analysis, and then group aggregated with chain P-technique. The composite pattern suggested a three-component solution consisting of cardiac rate, cardiac contractility, and peripheral resistance factors. Individual profiles were compared to the composite pattern; these profiles differed in the number of components derived, percentage variance explained by these components, and relative dominance of specific CV components. A hypothesis that emerged is that the subjects differed in the complexity of CV control. It appears that the idiodynamic framework, combined with novel research designs and statistical methods, may help expand the view of CV reactivity beyond the traditional unitary view as response magnitude. PMID- 15119380 TI - A laboratory animal model of human shift work. AB - The purpose of this study was to develop a laboratory animal model of human shift work. Two methods of monitoring circadian rhythms in rats were employed: an activity wheel cage, where number of wheel revolutions (WR) were counted, and an internal radio transmitter, which recorded gross motor activity (GMA) and body temperature (BT). Rats were implanted with biotelemetry transmitters that detected GMA and BT and were placed in activity wheel cages. A 12 hour/12 hour light/dark cycle was maintained. Subjects were subdivided into two groups: control and experimental. Following a habituation period of 15 days, in which animals had ad-libitum access to food and water and unlimited access to the running wheel, the experimental period ensued for 22 days. Control animals were food restricted and their activity wheels were locked during the light; experimental animals were food restricted and their activity wheels were locked during the dark. At the end of the experimental period, animals were returned to the habituation paradigm for 15 days. Recordings of WR, GMA and BT, as well as daily monitoring of body weight and food intake, indicated that experimental animals resembled humans employed in a shift work schedule. In the experiment, the light entrainable oscillator and the food entrainable oscillator were uncoupled in experimental animals, producing alterations in activity/rest cycles, consummatory behavior, and overt behavior. Since similar alterations occur in shift workers, it is proposed that the experimental paradigm presented in this manuscript is a useful model of shift work and provides a framework upon which future experiments may be conducted. PMID- 15119381 TI - Parts per trillion sensitivity for ethane in air with an optical parametric oscillator cavity leak-out spectrometer. AB - Spectroscopic detection of ethane in the 3-microm wavelength region was performed by means of a cw optical parametric oscillator and cavity leak-out. We achieved a minimum detectable absorption coefficient of 1.6 x 10(-10) cm 1/square root of Hz, corresponding to an ethane detection limit of 6 parts per trillion/square root of Hz. For 3-min integration time the detection limit was 0.5 parts per trillion. The levels are to our knowledge the best demonstrated so far. These frequency-tuning capabilities facilitated multigas analysis with simultaneous monitoring of ethane, methane, and water vapor in human breath. PMID- 15119382 TI - Partially coherent nonparaxial beams. AB - The concept of a partially coherent nonparaxial beam is proposed. A closed-form expression for the propagation of nonparaxial Gaussian Schell model (GSM) beams in free space is derived and applied to study the propagation properties of nonparaxial GSM beams. It is shown that for partially coherent nonparaxial beams a new parameter f(sigma) has to be introduced, which together with the parameter f, determines the beam nonparaxiality. PMID- 15119383 TI - High-efficiency broadband diffractive elements based on polarization gratings. AB - A method is introduced for designing paraxial-domain diffractive elements working over a broad frequency range. The method is based on space-variant manipulation of the state of polarization by form-birefringent binary diffractive structures. It is shown that any scalar phase transmission function can be realized by use of such polarization-modulating structures and that at least in some cases it is even possible to exceed the scalar paraxial-domain upper bounds of diffraction efficiency over a broad frequency band. PMID- 15119384 TI - Bandpass engineering of lithographically scribed channel-waveguide Bragg gratings. AB - We propose and demonstrate a powerful approach to spectral bandpass engineering (apodization) of one-dimensional channel-waveguide Bragg reflectors. Bandpass engineering is accomplished by precise photolithographic control of the length and the longitudinal placement of individual grating lines, which provides unique line-by-line diffractive amplitude and phase control. Channel-waveguide gratings that exhibit complex filtering functions have been fabricated and modeled. When a second-order apodization effect that comprises effective waveguide refractive index variation with grating-line length is included in the simulation, extraordinary agreement between predicted and observed spectral passband profiles is obtained. PMID- 15119385 TI - Bragg fiber design for linear polarization. AB - A new design is presented for Bragg fibers that allows low-loss propagation for linearly polarized light. Predictions based on a simple ray model show that approximately doubling the thickness of the first wall layer results in low losses at TM-like boundaries while keeping TE-like boundary losses manageable. This contrasts sharply with conventional quarter-wave designs that are extremely low loss for TE01 modes but very high loss for linear polarization. We fabricate Bragg fibers based on this design concept in a Si/SiO2 system and verify experimentally that they propagate linearly polarized light with losses less than 6 dB/cm over a 60-nm spectral range. PMID- 15119386 TI - Analysis of a widely tunable long-period grating by use of an ultrathin cladding layer and higher-order cladding mode coupling. AB - A widely tunable long-period grating in single-mode fiber is analyzed by use of an ultrathin cladding layer and higher-order cladding mode coupling. The numerical simulation shows that a 225-nm tuning range in the newly designed ultrathin long-period grating (cladding thickness, 35 microm) with third-order cladding mode coupling can be obtained. The analyzed tuning range is seven times wider than those of the other known long-period gratings. We believe that the proposed highly sensitive long-period grating will be widely used as a gain flattening filter for ultrawideband optical amplifiers and fast tunable filters in dynamic optical communication systems. PMID- 15119387 TI - Hydrogen loading of optical waveguides by use of host diluent gases. AB - Recent dramatic improvements in photosensitivity gas mixtures when very low hydrogen concentrations (as low as 0.1%) were used are shown to involve condensation of the host gas. The vapor pressure of the host determines the effective partial pressure of the hydrogen, and this accounts for the bulk of the observed enhancements. PMID- 15119388 TI - Small-core single-mode microstructured polymer optical fiber with large external diameter. AB - A preform sleeving technique is demonstrated that allows the fabrication of single-mode polymer microstructured fiber with the smallest core and hole dimensions yet reported to our knowledge. For a fixed triangular hole pattern a range of fibers is produced by adjustment to the operating conditions of the draw tower. Numerical modeling is carried out for one of the fibers produced with a 570-microm external diameter, a core diameter of 2.23 microm, an average hole diameter of 0.53 microm, and an average hole spacing of 1.38 microm. This fiber was shown to be endlessly single mode. PMID- 15119389 TI - Dispersion management for randomly varying optical fibers. AB - An asymptotic theory for optical pulse propagation in a dispersion-managed (DM) fiber with random dispersion is presented. The validity of the theory is verified with direct numerical simulation. The equations that describe the slow evolution of initial pulses have special solutions that, for fibers with moderate noise in the dispersion profile, perform much better than ideal DM solitons optimized for the unperturbed fiber. PMID- 15119390 TI - Detection and tracking of small moving objects in image sequences by use of nonlinear spatiotemporal optical systems. AB - We demonstrate the capability of localized structures of spatiotemporal nonlinear optical systems for detection and tracking of small moving objects. Such nonlinear dynamics based image-processing systems are readily implemented using an existing parallel optoelectronic architecture designed for real-time operation. PMID- 15119391 TI - Measurements of the indium hyperfine structure in an atmospheric-pressure flame by use of diode-laser-induced fluorescence. AB - We report on what we believe is the first demonstration of laser-induced fluorescence (LIF) in flames by use of diode lasers. Indium atoms seeded into an atmospheric-pressure flame at trace concentrations are excited by a blue GaN laser operating near 410 nm. The laser is mounted in an external-cavity configuration, and the hyperfine spectrum of the 5(2)P1/2 --> 6(2)S1/2 transition is captured at high resolution in single-wavelength sweeps lasting less than one tenth of a second. The research demonstrates the potential of diode-based LIF for practical diagnostics of high-temperature reactive flows. PMID- 15119392 TI - Room-temperature, continuous-wave 1-W green power by single-pass frequency doubling in a bulk periodically poled MgO:LiNbO3 crystal. AB - Continuous-wave high-power green light generation at room temperature is reported in a single-pass frequency-doubling configuration with bulk periodically poled MgO:LiNbO3 crystal placed outside a diode end-pumped Nd:GdVO4 laser. The MgO:LiNbO3 samples of 6.95-microm domain period, uniform periodicity, and 50% duty cycle along the entire crystal length are fabricated by use of a high voltage multipulse poling method. A maximum power of 1.18 W at 531 nm with 16.8% conversion efficiency is obtained from a 2-mm-thick, 25-mm-long MgO:LiNbO3 crystal; the corresponding internal green power and conversion efficiency are 1.38 W and 19.6%, respectively, whereas the normalized conversion efficiency is 3.3%/W. PMID- 15119393 TI - Laser oscillation with low quantum defect in Yb:GdVO4, a crystal with high thermal conductivity. AB - A Yb:GdVO4 single crystal was grown by the Czochralski process. Its thermal conductivity was measured by photothermal analysis and reached 8.1 and 7.1 W m( 1) K(-1) along and perpendicular to the c axis, respectively. These values are in good agreement with the predicted ones. The optical spectroscopy of the crystal, such as its absorption and emission cross sections under the two polarizations, its fluorescence lifetime, and its laser parameters, is investigated. Cw laser oscillation is obtained for what is to our knowledge the first time in this material under titanium sapphire pumping at 984 nm. We obtain 420 mW of output power for a 2% output coupler at 1029 nm. With a thin sample, the laser wavelength decreases to 1015 nm. In this case the quantum defect is as small as 2.9%. The weak heat release of the material suggests good prospects for its use in high-power applications. PMID- 15119394 TI - Lasing characteristics and optimizations of a diode-side-pumped Tm, Ho:GdVO4 laser. AB - A diode-pumped Tm, Ho:GdVO4 laser with a side-pumping configuration is demonstrated for the first time to our knowledge. Optimum Tm and Ho dopant concentrations for GdVO4 are somewhat lower than those for garnet and fluoride crystals. With a 3% Tm, 0.3% Ho:GdVO4 crystal an output energy of 31.2 mJ and a slope efficiency of 14.5% were obtained in normal-mode operation at room temperature. PMID- 15119395 TI - Eigenstates and eigenenergies of four-wave-mixing models. AB - We present explicit analytical expressions of all the eigenenergies and eigenstates for two different kinds of four-wave-mixing model in terms of a parameter lambda without the so-called Bethe ansatz assumption. The parameter lambda is shown to be determined by the roots of a simple polynomial. We also obtain the exact explicit analytical expressions of infinite eigenstates and energies without any unknown parameter. PMID- 15119396 TI - Broadband source generated by stimulated Raman scattering and four-wave mixing in a highly nonlinear optical fiber ring cavity. AB - A novel high-power broadband source based on the combined action of stimulated Raman scattering and parametric four-wave mixing in a highly nonlinear dispersion shifted fiber ring cavity is investigated experimentally. An output spectrum of 1510-1580 nm with a stable power of 91 mW is demonstrated with a dual-wavelength pumping scheme. PMID- 15119397 TI - Dissipative photonic lattice solitons. AB - We show that discrete dissipative optical lattice solitons are possible in waveguide array configurations that involve periodically patterned semiconductor optical amplifiers and saturable absorbers. The characteristics of these low power soliton states are investigated, and their propagation constant eigenvalues are mapped on Floquet-Bloch band diagrams. The prospect of observing such low power dissipative lattice solitons is discussed in detail. PMID- 15119398 TI - Highly efficient narrow-line generation by difference-frequency mixing of a green pump and the stokes seed in RbTiOPO4 crystals: excitation of 943-nm emission. AB - An efficient and compact scheme for diode-pumped Nd:YLF laser wavelength conversion to 943 nm was demonstrated by use of difference-frequency mixing and stimulated Raman scattering. We believe that this is the highest conversion efficiency from the laser fundamental wavelength reported to date. It is shown that RbTiOPO4 crystals are capable of providing highly efficient frequency mixing as a nonlinear medium. PMID- 15119399 TI - Cutoff solitons in axially uniform systems. AB - The optical response of axially uniform nonlinear photonic bandgap fibers is studied theoretically. We observe gap-soliton-like generation and associated bistability, similar to what is typically found in periodically modulated nonlinear structures. This response stems from the nature of the guided-mode dispersion relations, which involve a frequency cutoff at zero wave vector. In such systems, solutions with zero group velocities and minimal coupling to radiation modes come in naturally. We term such solitons "cutoff solitons"; they provide an interesting alternative to gap solitons in periodically index modulated fibers for in-fiber all-optical signal processing. PMID- 15119400 TI - Controlling image size as a function of distance and wavelength in Fresnel transform reconstruction of digital holograms. AB - A method for controlling the size of amplitude and phase images reconstructed from digital holograms by the Fresnel-transform method is proposed and demonstrated. The method can provide a constant reconstruction pixel width in the reconstructed image plane, independent of the recording and reconstruction distance. The proposed method makes it possible to maintain the size of an object for a sequence of digital holograms recorded at different distances and, therefore, to subtract phase maps for an object recorded at different distances. Furthermore, the method solves the problem of superimposition in multiwavelength digital holography for color display and holographic interferometry applications. PMID- 15119401 TI - Digital microscopy using phase-shifting digital holography with two reference waves. AB - A lensless, coherent optical microscope is described that uses a version of phase shifting digital holography (PSDH) in conjunction with a field backpropagation algorithm to form coherent images of transmission-type objects. The PSDH is implemented by use of only two reference waves, in contrast with the usual implementation that requires four quadrature phase-shifting reference waves. Therefore only two digital holograms need to be recorded, and the complexity of the microscopic system is reduced. Experimental results are presented that compare images generated from conventional Gabor digital holography, two reference-wave PSDH, and conventional white-light microscopy. PMID- 15119402 TI - Reducing the out-of-plane radiation loss of photonic crystal waveguides on high index substrates. AB - Two-dimensional photonic crystal linear defect waveguides on semiconductor substrates are studied. It is predicted that the out-of-plane radiation loss can be reduced by shifting one side of the photonic crystal cladding by one-half period with respect to the other along the propagation direction. PMID- 15119403 TI - Design and optimization of one-dimensional photonic crystals for thermophotovoltaic applications. AB - We explore the optical characteristics and fundamental limitations of one dimensional (1D) photonic crystal (PhC) structures as means for improving the efficiency and power density of thermophotovoltaic (TPV) and microthermophotovoltaic (MTPV) devices. We analyze the optical performance of 1D PhCs with respect to photovoltaic diode efficiency and power density. Furthermore, we present an optimized dielectric stack design that exhibits a significantly wider stop band and yields better TPV system efficiency than a simple quarter-wave stack. The analysis is done for both TPV and MTPV devices by use of a unified modeling framework. PMID- 15119404 TI - Refractive microlens array for wave-front analysis in the medium to hard x-ray range. AB - We report an alternative approach to x-ray wave-front analysis that uses a refractive microlens array as a Shack-Hartmann sensor. The sensor was manufactured by self-assembly and electroplating techniques and is suitable for high-resolution wave-front analysis of medium to hard x rays. We demonstrate its effectiveness at an x-ray energy of 3 keV for analysis of x-ray wave-front perturbations caused by microscopic objects. The sensor has potential advantages over other methods for x-ray phase imaging and will also be useful for the characterization of x-ray beams and optics. PMID- 15119405 TI - Optical amplification in a first-generation dendritic organic semiconductor. AB - We report a study of a new class of organic semiconductor as an optical gain medium. We demonstrate amplification of violet light by use of stimulated emission in a solution of a first-generation bis-fluorene-cored semiconducting dendrimer. Amplification is also observed in the solid state by means of amplified spontaneous emission in an optically pumped dendrimer planar waveguide. Gains of 36 dB cm(-1) at 420 nm and 26 dB cm(-1) at 390 nm in solution and 350 dB cm(-1) in the solid state are obtained. These results show that semiconducting dendrimers have potential as visible laser and amplifier materials. PMID- 15119406 TI - Goos-Hanchen shifts at the interfaces between left- and right-handed media. AB - The Goos-Hanchen shift caused by total internal reflection at the interface between two media is analyzed. For two media of the same handedness the Goos Hanchen phase shift opposes the phase variation associated with propagation through the incident medium. The Goos-Hanchen lateral shift is in the same direction as the horizontal component of the incident energy flux. Conversely, for two media of opposite handedness the Goos-Hanchen phase shift reinforces the phase variation associated with propagation through the incident medium. The lateral shift is in the opposite direction of the horizontal component of the incident energy flux. PMID- 15119407 TI - Polarization singularities in optical lattices. AB - Polarization singularities are shown to be unavoidable features of three dimensional optical lattices. These singularities take the form of lines of circular polarization, C lines, and lines of linear polarization, L lines. The polarization figures surrounding a C line (L line) rotate about the line with winding number +/-1/2 (+/-1). C and L lines permeate the lattice, meander throughout the unit cell, and form closed loops. Surprisingly, every point in a linearly polarized optical lattice is found to be a singularity about which the surrounding polarization vectors rotate with an integer winding number. PMID- 15119408 TI - Polarization-sensitive effects of solgel materials containing various chiral media. AB - The polarization-sensitive effects of solgel materials containing various chiral media were measured experimentally. The results show that the solgel material displays optical activity when it contains organic chiral molecules and manifests depolarization when it contains inorganic chiral microcrystals with a particle size of 70 microm. Solgel material containing glass powder that also has a particle size of 70 microm displays a polarization held characteristic (i.e., the polarization of the output light is the same as that of the input light). PMID- 15119409 TI - Transverse spatial coherence of a transient nickellike silver soft-x-ray laser pumped by a single picosecond laser pulse. AB - The degree of spatial coherence in the direction perpendicular to the target surface is reported for a transient nickellike silver x-ray laser at 13.9 nm. An x-ray laser plasma column was produced by irradiating a slab silver target with a single shaped picosecond laser pulse with energy less than 3 J. Young's double slit method was applied to measure the fringe visibility as a function of the slit separation for different target lengths. The diameter of the equivalent incoherent source and the coherence radius of the output radiation were determined as well. PMID- 15119410 TI - Tunable two-dimensional femtosecond spectroscopy. AB - We have developed a two-dimensional (2D) Fourier-transform femtosecond spectroscopy technique for the visible spectral region. Three-pulse photon echo signals are generated in a phase-matched noncollinear four-wave mixing box geometry that employs a 3-kHz repetition-rate laser system and optical parametric amplification. Nonlinear signals are fully characterized in amplitude and phase by spectral interferometry. Unlike for previous setups, we achieve long-term phase stability by employing diffractive optics and interferometric accuracy of excitation-pulse time delays by using movable glass wedges. As an example of this technique, 2D correlation and relaxation spectra at 600 nm are shown for a solution of Nile Blue dye in acetonitrile. PMID- 15119411 TI - Optical light bullets in a pure Kerr medium. AB - We show that small negative fourth-order dispersion can arrest spatiotemporal collapse of ultrashort pulses with anomalous dispersion in a planar waveguide with pure Kerr nonlinearity, resulting in (2 + 1)D optical bullets. Similarly to solitons, these bullets undergo elastic collisions. Since these bullets can self trap from noisy Gaussian input beams and propagate without any power losses, this result may be used to realize experimentally stable, nondissipative optical bullets. PMID- 15119412 TI - Femtosecond pulse-shape modulation at kilohertz rates. AB - We demonstrate a new scanning femtosecond pulse-shaping technique that allows pulse shapes to be modulated at kilohertz rates. This technique is particularly useful for lock-in measurements in which the signal is synchronized with the alternating pulse shapes. The pulse-shape lock-in technique is demonstrated in resonant coherent anti-Stokes Raman scattering, where it is shown to significantly improve the ratio of the resonant signal to both the nonresonant background and to noise. PMID- 15119413 TI - High-order harmonic generation by nonlinear reflection of an intense high contrast laser pulse on a plasma. AB - We demonstrate the use of a plasma mirror to obtain 60-fs 10-TW laser pulses with a temporal contrast of 10(8) on a nanosecond time scale and 10(6) on a picosecond time scale, and we use these high-contrast pulses to generate high harmonics by nonlinear reflection on a plasma with a steep electronic density gradient. Well collimated harmonics up to 20th order are observed for a laser intensity of approximately equal to 3 x 10(17) W/cm2, whereas no harmonics are obtained without the plasma mirror. PMID- 15119414 TI - Terahertz transmission properties of thin, subwavelength metallic hole arrays. AB - We present experimental results of the transmission magnitude and phase change of terahertz pulses through thin metallic films patterned with subwavelength hole arrays on silicon wafers. Terahertz time-domain spectroscopy measurements reveal a sharp phase peak centered on the surface plasmon resonance. Correspondingly, and consistent with the Kramers-Kronig relations, the measured transmission magnitude has the shape of the derivative of this peak. In addition, we determine that the aperture shape has a notable effect on the transmission properties of two-dimensional hole arrays. PMID- 15119415 TI - Direct measurement of wave-front distortion induced during second-harmonic generation: application to breakup-integral compensation. AB - The wave-front distortion of femtosecond laser pulses recorded with a Shack Hartmann analyzer makes it possible to retrieve the nonlinear index of refraction of different glasses and the nonlinear phase shift induced during second-harmonic generation in beta-barium borate (BBO) crystal versus the phase mismatch. It is shown that the nonlinear phase shift induced in a 2-mm-thick BBO crystal allows compensation for up to a 2pi breakup-integral induced in a 4-cm fused-silica glass. The stability of the compensation is reported to be from 10 to 100 GW cm( 2). PMID- 15119416 TI - The power of self-motivation. PMID- 15119417 TI - A commitment to compassion at the end of life. PMID- 15119418 TI - An overview of cancer in the older adult. AB - With the number of elders rising and the incidence of cancer increasing with age, a discussion of cancer in older adults is critical for medical-surgical nurses. An understanding of how aging affects cancer incidence, etiology, biology, diagnosis, treatment, and patient response will help medical-surgical nurses provide quality, comprehensive care to elders with cancer. This includes the ability to assess age-related limitations and initiate intervention strategies as appropriate for older adults with cancer. PMID- 15119419 TI - Informed nursing practice: the administration of oxygen to patients with COPD. AB - Although the Hypoxic Drive Theory has been a long-time influence on the care of patients with chronic obstructive pulmonary disease (COPD), in reality a relatively fine line exists between optimal and excessive oxygen therapy. Medical surgical nurses need a clear understanding of the benefits and the possible complications of oxygen therapy in patients with COPD. PMID- 15119420 TI - Ethical guidelines for assisting patients with end-of-life decision making. AB - The medical-surgical nurse must be prepared to care for patients at the end of life as they face a multitude of complex issues surrounding physical, psychological, and spiritual needs. End-of-life care must not only control symptoms, but also address the social, psychological, and spiritual needs of the patient. By using a decision framework, the nurse will be better prepared to assist patients in exploring options for end-of-life care. PMID- 15119421 TI - Consumer health information on the Web: trends, issues, and strategies. AB - Medical-surgical nurses are at the forefront of the current trends in patient information and choice in medical care. The trends in health information access and quality, and decision support strategies are reviewed. Valuable online resources for nurses to share with their patients and families will be presented and applied using a case study. Elder health is used to illustrate these issues. PMID- 15119422 TI - HIV disease susceptibility in women and the barriers to adherence. AB - The late 1990s saw a decrease in the total number of new cases of HIV (human immunodeficiency virus) and AIDS (acquired immunodeficiency syndrome). However, data often do not reflect the increased incidence of HIV in women, particularly women from minority groups. The purposes of this clinical review are to address the disease susceptibility and disease manifestations of HIV in women, and to discuss selected barriers to adherence to antiretroviral medications such as stigma, social support, and depression. PMID- 15119423 TI - Use of the insertable loop recorder to detect cardiac arrhythmias during syncopal episodes. AB - The Reveal Plus insertable loop recorder (ILR) is an implantable device used to detect cardiac arrhythmias in patients suffering from undiagnosed syncope. It has a high diagnostic yield, is easily inserted, and results in better patient outcomes. As medical-surgical nurses begin to see more patients with ILRs, they need an understanding of device use and care. PMID- 15119424 TI - Cancer screening in women with chronic illness: the unanswered questions. AB - The literature offers conflicting results regarding whether the cancer screening needs of women with chronic illness are being met. Cancer screening in this population of women is in need of further investigation to address its cost effectiveness, the health care system's priority and management of screening the risks of cancer-related illness in women with co-morbidities, and the perceptions of the affected women. While other care providers are often focused on disease management, nurses who pride themselves with providing holistic care can take an active role in ensuring all women receive preventive health screening. PMID- 15119425 TI - The confluence of two clinical specialties: genetics and assisted reproductive technologies. AB - The confluence of genetic and reproductive technologies has significantly expanded the scope of available reproductive options. To ensure appropriate dissemination of information and timely referral for preconception counseling, nurses in all specialties must be aware of the clinical indications for advanced reproductive technologies. PMID- 15119426 TI - Sexuality. PMID- 15119427 TI - Preparing for certification: test-taking strategies. PMID- 15119428 TI - Military medical-surgical nurses in operations other than war: MEDCAP missions during deployments. PMID- 15119429 TI - Enteral nutrition: an update on practice recommendations. PMID- 15119430 TI - Evolution of multihost parasites. AB - Multihost parasites can infect different types of hosts or even different host species. Epidemiological models have shown the importance of the diversity of potential hosts for understanding the dynamics of infectious disease (e.g., the importance of reservoirs), but the consequences of this diversity for virulence and transmission evolution remain largely overlooked. Here, I present a general theoretical framework for the study of life-history evolution of multihost parasites. This analysis highlights the importance of epidemiology (the relative quality and quantity of different types of infected hosts) and between-trait constraints (both within and between different hosts) to parasite evolution. I illustrate these effects in different transmission scenarios under the simplifying assumption that parasites can infect only two types of hosts. These simple but contrasted evolutionary scenarios yield new insights into virulence evolution and the evolution of transmission routes among different hosts. Because many of the pathogens that have large public-health and agricultural impacts have complex life cycles, an understanding of their evolutionary dynamics could hold substantial benefits for management. PMID- 15119431 TI - The diffusive spread of alleles in heterogeneous populations. AB - The spread of genes and individuals through space in populations is relevant in many biological contexts. I study, via systems of reaction-diffusion equations, the spatial spread of advantageous alleles through structured populations. The results show that the temporally asymptotic rate of spread of an advantageous allele, a kind of invasion speed, can be approximated for a class of linear partial differential equations via a relatively simple formula, c = 2 square root of (rD), that is reminiscent of a classic formula attributed to R. A. Fisher. The parameters r and D represent an asymptotic growth rate and an average diffusion rate, respectively, and can be interpreted in terms of eigenvalues and eigenvectors that depend on the population's demographic structure. The results can be applied, under certain conditions, to a wide class of nonlinear partial differential equations that are relevant to a variety of ecological and evolutionary scenarios in population biology. I illustrate the approach for computing invasion speed with three examples that allow for heterogeneous dispersal rates among different classes of individuals within model populations. PMID- 15119432 TI - Power and potential bias in field studies of natural selection. AB - The advent of multiple regression analyses of natural selection has facilitated estimates of both the direct and indirect effects of selection on many traits in numerous organisms. However, low power in selection studies has possibly led to a bias in our assessment of the levels of selection shaping natural populations. Using calculations and simulations based on the statistical properties of selection coefficients, we find that power to detect total selection (the selection differential) depends on sample size and the strength of selection relative to the opportunity of selection. The power of detecting direct selection (selection gradients) is more complicated and depends on the relationship between the correlation of each trait and fitness and the pattern of correlation among traits. In a review of 298 previously published selection differentials, we find that most studies have had insufficient power to detect reported levels of selection acting on traits and that, in general, the power of detecting weak levels of selection is low given current study designs. We also find that potential publication bias could explain the trend that reported levels of direct selection tend to decrease as study sizes increase, suggesting that current views of the strength of selection may be inaccurate and biased upward. We suggest that studies should be designed so that selection is analyzed on at least several hundred individuals, the total opportunity of selection be considered along with the pattern of selection on individual traits, and nonsignificant results be actively reported combined with an estimate of power. PMID- 15119433 TI - Multilevel selection and the partitioning of covariance: a comparison of three approaches. AB - Where the evolution of a trait is affected by selection at more than one hierarchical level, it is often useful to compare the magnitude of selection at each level by asking how much of the total evolutionary change is attributable to each level of selection. Three statistical partitioning techniques, each designed to answer this question, are compared, in relation to a simple multilevel selection model in which a trait's evolution is affected by both individual and group selection. None of the three techniques is wholly satisfactory: one implies that group selection can operate even if individual fitness is determined by individual phenotype alone, whereas the other two imply that group selection can operate even if there is no variance in group fitness. This has significant implications both for our understanding of what the term "multilevel selection" means and for the traditional concept of group selection. PMID- 15119434 TI - Evolutionary dynamics of complex biomechanical systems: an example using the four bar mechanism. AB - Like many phenotypic traits, biomechanical systems are defined by both an underlying morphology and an emergent functional property. The relationship between these levels may have a profound impact on how selection for functional performance is translated into morphological evolution. In particular, complex mechanical systems are likely to be highly redundant, because many alternative morphologies yield equivalent functions. We suggest that this redundancy weakens the relationship between morphological and functional diversity, and we illustrate this effect using an evolutionary model of the four-bar lever system of labrid fishes. Our results demonstrate that, when traits are complex, the morphological diversity of a clade may only weakly predict its mechanical diversity. Furthermore, parallel or convergent selection on function does not necessarily produce convergence in morphology. Empirical observations suggest that this weak form-function relationship has contributed to the morphological diversity of labrid fishes, as functionally equivalent species may nevertheless possess morphologically distinct jaws. We suggest that partial decoupling of morphology and mechanics due to redundancy is a major factor in morphological diversification. PMID- 15119435 TI - Variational and genetic properties of developmental stability in Dalechampia scandens. AB - Because low developmental stability may compromise the precision with which adaptations can be reached, the variability and genetic basis of developmental stability are important evolutionary parameters. Developmental stability is also an important clue to understanding how traits are regulated to achieve their phenotypic target value. However, developmental stability must be studied indirectly through proxy variables, such as fluctuating asymmetry, that are suggested to have noisy and often nonlinear relationships to the underlying variable of interest. In this paper we first show that mean-standardized measures of variance and covariance in fluctuating asymmetry, unlike heritabilities, repeatabilities, and correlations, are linearly related to corresponding measures of variation in underlying developmental stability. We then examine the variational properties of developmental stability in a population of the Neotropical vine, Dalechampia scandens (Euphorbiaceae). By studying fluctuating asymmetry in a large number of floral characters in both selfed and outcrossed individuals in a diallel design, we assemble strong evidence that both additive genetic and individual variation and covariation in developmental stability are virtually absent in this population. PMID- 15119436 TI - An experimental test of the adaptive evolution of phototropins: blue-light photoreceptors controlling phototropism in Arabidopsis thaliana. AB - Phototropins are blue-light photoreceptor molecules mediating the capacity for phototropism or bending toward or away from directional light. Like the red-light sensing phytochromes that control shade avoidance, phototropins modulate developmental plasticity in plant architecture. Yet, unlike phytochromes, the adaptive significance of phototropins has been largely a topic of conjecture. In Arabidopsis thaliana, phototropism of seedling and plant stems is under the control of two paralogous genes, PHOT1 and PHOT2, that encode different phototropins with partially redundant light response qualities. The PHOT1 gene product interacts with the NPH3 gene product to cause phototropic bending over a broad range of light intensity, from very weak light in the soil to stronger light in the aerial environment. The PHOT2 gene product modulates shoot bending in response to light of higher intensity only. We compared the fitness of wild type, phot1, phot2, and nph3 genotypes over a range of light conditions in the field. Seeds were sown in the field on the soil surface and left bare or covered with either gravel or bark mulch chips. Plantings were made under full sun and dense canopy cover. Rates of seedling emergence, survival to flowering, and total seed set were measured. All mutant genotypes had significantly reduced lifetime fitness compared to wild-type. Consistent with their different fluence rate sensitivities, phot1 and phot2 signaling pathways affected fitness at discrete life-cycle stages. Fitness costs of phot1 and nph3 were expressed mainly during seedling emergence from the soil whereas that of phot2 was expressed solely after emergence. Surprisingly, the only significant genotype-by-environment interaction for fitness occurred during emergence: genotypes blind to dim blue light (phot1 and nph3) had poor emergence in the open, but not in the shade. Possibly, the loss of negative phototropism in seedling roots of mutant genotypes reduced establishment success in open (dry soil) conditions. Results show that phototropin-modulated pathways are adaptive and that their evolution has involved functional specialization. However, mechanism(s) of selection on these pathways remain a mystery. PMID- 15119437 TI - Heterochronic developmental shift caused by thyroid hormone in larval sand dollars and its implications for phenotypic plasticity and the evolution of nonfeeding development. AB - Recent work on a diverse array of echinoderm species has demonstrated, as is true in amphibians, that thyroid hormone (TH) accelerates development to metamorphosis. Interestingly, the feeding larvae of several species of sea urchins seem to obtain TH through their diet of planktonic algae (exogenous source), whereas nonfeeding larvae of the sand dollar Peronella japonica produce TH themselves (endogenous source). Here we examine the effects of TH (thyroxine) and a TH synthesis inhibitor (thiourea) on the development of Dendraster excentricus, a sand dollar with a feeding larva. We report reduced larval skeleton lengths and more rapid development of the juvenile rudiment in the exogenous TH treatments when compared to controls. Also, larvae treated with exogenous TH reached metamorphic competence faster at a significantly reduced juvenile size, representing the greatest reduction in juvenile size ever reported for an echinoid species with feeding larvae. These effects of TH on D. excentricus larval development are strikingly similar to the phenotypically plastic response of D. excentricus larvae reared under high food conditions. We hypothesize that exogenous (algae-derived) TH is the plasticity cue in echinoid larvae, and that the larvae use ingested TH levels as an indicator for larval nutrition, ultimately signaling the attainment of metamorphic competence. Furthermore, our experiments with the TH synthesis inhibitor thiourea indicate that D. excentricus larvae can produce some TH endogenously. Endogenous TH production might, therefore, be a shared feature among sand dollars, facilitating the evolution of nonfeeding larval development in that group. Mounting evidence on the effects of thyroid hormones in echinoderm development suggests life history models need to incorporate metamorphic hormone effects and the evolution of metamorphic hormone production. PMID- 15119438 TI - Phylogenetic and molecular evidence for allochronic speciation in gall-forming aphids (Pemphigus). AB - Sympatric populations can diverge when variation in phenology or life cycle causes them to mate at distinctly different times. We report patterns consistent with this process (allochronic speciation) in North American gall-forming aphids, in the absence of a host or habitat shift. Pemphigus populi-transversus Riley and P. obesinymphae Aoki form a monophyletic clade within the North American Pemphigus group. They are sympatric on the eastern cottonwood, Populus deltoides (Salicaceae), but have distinctly different life cycles, with sexual stages offset by approximately six months. Field evidence indicates that intermediate phenotypes do not commonly occur, and mitochondrial and bacterial endosymbiont DNA sequences show no maternal gene flow between the two species. Because a genetically distinct population of P. obesinymphae occurs in the southwestern United States on Populus fremontii, we consider the possibility of an initial allopatric phase in the divergence. We discuss the likely origins of the host use patterns in P. obesinymphae, and the larger sequence of evolutionary changes that likely led to the sympatric divergence of P. populi-transversus and P. obesinymphae. A plausible interpretation at this stage of investigation is that a shift in timing of the life cycle in an ancestral population, correlated with an underlying phenological complexity in its host plant, spurred divergence between the incipient species. PMID- 15119439 TI - Codiversification in an ant-plant mutualism: stem texture and the evolution of host use in Crematogaster (Formicidae: Myrmicinae) inhabitants of Macaranga (Euphorbiaceae). AB - We investigate the evolution of host association in a cryptic complex of mutualistic Crematogaster (Decacrema) ants that inhabits and defends Macaranga trees in Southeast Asia. Previous phylogenetic studies based on limited samplings of Decacrema present conflicting reconstructions of the evolutionary history of the association, inferring both cospeciation and the predominance of host shifts. We use cytochrome oxidase I (COI) to reconstruct phylogenetic relationships in a comprehensive sampling of the Decacrema inhabitants of Macaranga. Using a published Macaranga phylogeny, we test whether the ants and plants have cospeciated. The COI phylogeny reveals 10 well-supported lineages and an absence of cospeciation. Host shifts, however, have been constrained by stem traits that are themselves correlated with Macaranga phylogeny. Earlier lineages of Decacrema exclusively inhabit waxy stems, a basal state in the Pachystemon clade within Macaranga, whereas younger species of Pachystemon, characterized by nonwaxy stems, are inhabited only by younger lineages of Decacrema. Despite the absence of cospeciation, the correlated succession of stem texture in both phylogenies suggests that Decacrema and Pachystemon have diversified in association, or codiversified. Subsequent to the colonization of the Pachystemon clade, Decacrema expanded onto a second clade within Macaranga, inducing the development of myrmecophytism in the Pruinosae group. Confinement to the aseasonal wet climate zone of western Malesia suggests myrmecophytic Macaranga are no older than the wet forest community in Southeast Asia, estimated to be about 20 million years old (early Miocene). Our calculation of COI divergence rates from several published arthropod studies that relied on tenable calibrations indicates a generally conserved rate of approximately 1.5% per million years. Applying this rate to a rate-smoothed Bayesian chronogram of the ants, the Decacrema from Macaranga are inferred to be at least 12 million years old (mid-Miocene). However, using the extremes of rate variation in COI produces an age as recent as 6 million years. Our inferred timeline based on 1.5% per million years concurs with independent biogeographical events in the region reconstructed from palynological data, thus suggesting that the evolutionary histories of Decacrema and their Pachystemon hosts have been contemporaneous since the mid-Miocene. The evolution of myrmecophytism enabled Macaranga to radiate into enemy-free space, while the ants' diversification has been shaped by stem traits, host specialization, and geographic factors. We discuss the possibility that the ancient and exclusive association between Decacrema and Macaranga was facilitated by an impoverished diversity of myrmecophytes and phytoecious (obligately plant inhabiting) ants in the region. PMID- 15119440 TI - Vibrational communication and reproductive isolation in the Enchenopa binotata species complex of treehoppers (Hemiptera: Membracidae). AB - Sexual communication can contribute to population divergence and speciation because of its effect on assortative mating. We examined the role of communication in assortative mating in the Enchenopa binotata species complex of treehoppers. These plant-feeding insects are a well studied case of sympatric speciation resulting from shifts to novel host-plant species. Shifting to hosts with different phenologies causes changes in life-history timing. In concert with high host fidelity, these changes reduce gene flow between populations on ancestral and novel hosts and facilitate a rapid response to divergent natural selection. However, some interbreeding can still occur because of partial overlap of mating periods. Additional behavioral mechanisms resulting in reproductive isolation may thus be important for divergence. In E. binotata, mating pairs form after an exchange of plant-borne vibrational signals. We used playback experiments to examine the relevance of inter- and intraspecific variation in male advertisement signals for female mate choice in a member of the E. binotata species complex. Female signals given in response to male signals provided a simple and reliable assay. Male species and male individual identity were important determinants of female responses. Females failed to respond to the signals of the two most closely related species in the complex, but they responded strongly to the signals of conspecific males, as well as to those of the most basal species in the complex. Communication systems in the E. binotata species complex can therefore play a role in reproductive isolation. Female responses were influenced by among-individual variation in male signals and females, suggesting the involvement of sexual selection in the evolution of these communication systems. PMID- 15119441 TI - Parasitism increases and decreases the costs of insecticide resistance in mosquitoes. AB - Adaptations conferring resistance to xenobiotics (antibiotics, insecticides, herbicides, etc.) are often costly to the organism's fitness in the absence of the selecting agent. In such conditions, and unless other mutations compensate for the costs of resistance, sensitive individuals are expected to out-reproduce resistant individuals and drive resistance alleles to a low frequency, with the rate and magnitude of this decline being proportional to the costs of resistance. However, this evolutionary dynamic is open to modification by other sources of selection acting on the relative fitness of susceptible and resistant individuals. Here we show parasitism not only as a source of selection capable of modifying the costs of organophosphate insecticide resistance in mosquitoes, but also that qualitatively different interactions (increasing or decreasing the relative fitness of resistant individuals) occurred depending on the particular form of resistance involved. As estimates of the parasite's fitness also varied according to its host's form of resistance, our data illustrate the potential for epidemiological feedbacks to influence the strength and direction of selection acting on resistance mutations in untreated environments. PMID- 15119442 TI - Ecological genetics of abdominal pigmentation in Drosophila falleni: a pleiotropic link to nematode parasitism. AB - Drosophila falleni belongs to the quinaria species group, whose species vary considerably in patterns of wing and abdominal pigmentation. Drosophila falleni itself exhibits substantial variation among wild flies in abdominal spotting patterns. A selection experiment revealed that natural populations of D. falleni harbor high levels of genetic variation for spot number: in 10 generations of selection modal spot number within populations declined from 18 (the modal number in wild-caught females) to as low as zero. Rearing flies at different temperatures shows that some of the variation among wild flies is likely to reflect variation in the environmental conditions under which they developed. Fitness assays did not reveal any cost of reduced spot number with respect to development time, adult survival, or female fecundity. However, spotless flies were almost twice as susceptible to infection by the nematode parasite Howardula aoronymphium. Thus, selection exerted by nematode parasites may influence pigmentation patterns and other, genetically correlated traits in natural populations D. falleni. PMID- 15119443 TI - Sexual selection for size and symmetry in a diversifying secondary sexual character in Drosophila bipectinata Duda (Diptera: Drosophilidae). AB - Results of intrapopulation studies of sexual selection and genetic variation and covariation underlying elements of the sex comb of Drosophila bipectinata are presented. The magnitude of the sex comb, a sexual ornament, varies significantly among Australasian populations, motivating research into the evolutionary mechanisms responsible for its incipient diversification. The comb is composed of stout black teeth on the front legs of males arranged in three distinct segments: C1, C2, and C3. Significant sexual selection in field populations in northeastern Queensland, Australia, was detected for increasing C2 and body size, and simultaneously for reducing comb positional fluctuating asymmetry. In contrast, sexual selection was not detected for other comb segments, nor for sternopleural bristle number or symmetry. Selection intensities for C2 and comb positional fluctuating asymmetry were similar in magnitude, and although they were opposite in sign, values across twelve sampling dates, or selection episodes, were uncorrelated. Heritability estimates for C2 were high and significant across years, whereas heritability estimates for comb positional asymmetry were small, and generally nonsignificant. The major sex comb segments (C1 and C2) were significantly and positively correlated genetically, indicating the potential for correlated evolution of these components of the comb under sexual selection. The original finding of a significant positive genetic correlation between the magnitude of this sex trait and its positional asymmetry indicates that the counteracting and independent selection pressures detected could contribute to the maintenance of genetic variation sustaining sexual selection. The study documents the simultaneous presence of sexual selection in nature and of heritable genetic variation underlying expression of the sex comb, fundamental conditions necessary for its adaptive diversification. Drosophila bipectinata may be a valuable model for studies of adaptive diversification and incipient speciation by sexual selection. PMID- 15119444 TI - Can intraspecific competition drive disruptive selection? An experimental test in natural populations of sticklebacks. AB - Theory suggests that frequency-dependent resource competition will disproportionately impact the most common phenotypes in a population. The resulting disruptive selection forms the driving force behind evolutionary models of niche diversification, character release, ecological sexual dimorphism, resource polymorphism, and sympatric speciation. However, there is little empirical support for the idea that intraspecific competition generates disruptive selection. This paper presents a test of this theory, using natural populations of the three-spine stickleback, Gasterosteus aculeatus. Sticklebacks exhibit substantial individual specialization associated with phenotypic variation and so are likely to experience frequency-dependent competition and hence disruptive selection. Using body size and relative gonad mass as indirect measures of potential fecundity and hence fitness, I show that an important aspect of trophic morphology, gill raker length, is subject to disruptive selection in one of two natural lake populations. To test whether this apparent disruptive selection could have been caused by competition, I manipulated population densities in pairs of large enclosures in each of five lakes. In each lake I removed fish from one enclosure and added them to the other to create paired low- and high-population-density treatments with natural phenotype distributions. Again using indirect measures of fitness, disruptive selection was consistently stronger in high-density than low-density enclosures. These results support long-standing theoretical arguments that intraspecific competition drives disruptive selection and thus may be an important causal agent in the evolution of ecological variation. PMID- 15119445 TI - Phylogenetic comparative analysis of life-history variation among populations of the lizard Sceloporus undulatus: an example and prognosis. AB - Over the past 15 years, phylogenetic comparative methods (PCMs) have become standard in the study of life-history evolution. To date, most studies have focused on variation among species or higher taxonomic levels, generally revealing the presence of significant phylogenetic effects as well as residual variation potentially attributable to adaptive evolution. Recently, population level phylogenetic hypotheses have become available for many species, making it possible to apply PCMs directly to the level at which experiments are typically used to test adaptive hypotheses. In this study, we present the results of PCMs applied to life-history variation among populations of the widespread and well studied lizard Sceloporus undulatus. Using S. undulatus (which may represent four closely related species) as an example, we explore the benefits of using PCMs at the population level, as well as consider the importance of several thorny methodological problems including but not limited to nonindependence of populations, lack of sufficient variation in traits, and the typically small sample sizes dictated by the difficulty of collecting detailed demographic data. We show that phylogenetic effects on life-history variation among populations of S. undulatus appear to be unimportant, and that several classic trade-offs expected by theory and revealed by many interspecific comparisons are absent. Our results suggest that PCMs applied to variation in life-history traits below the species level may be of limited value, but more studies like ours are needed to draw a general conclusion. Finally, we discuss several outstanding problems that face studies seeking to apply PCMs below the species level. PMID- 15119446 TI - Climatic and temporal effects on the expression of secondary sexual characters: genetic and environmental components. AB - Despite great interest in sexual selection, relatively little is known in detail about the genetic and environmental determinants of secondary sexual characters in natural populations. Such information is important for determining the way in which populations may respond to sexual selection. We report analyses of genetic and large-scale environmental components of phenotypic variation of two secondary sexual plumage characters (forehead and wing patch size) in the collared flycatcher Ficedula albicollis over a 22-year period. We found significant heritability for both characters but little genetic covariance between the two. We found a positive association between forehead patch size and a large-scale climatic index, the North Atlantic Oscillation (NAO) index, but not for wing patch. This pattern was observed in both cross-sectional and longitudinal data suggesting that the population response to NAO index can be explained as the result of phenotypic plasticity. Heritability of forehead patch size for old males, calculated under favorable conditions (NAO index > or = median), was greater than that under unfavorable conditions (NAO index < median). These changes occurred because there were opposing changes in additive genetic variance (VA) and residual variance (VR) under favorable and unfavorable conditions, with VA increasing and VR decreasing in good environments. However, no such effect was detected for young birds, or for wing patch size in either age class. In addition to these environmental effects on both phenotypic and genetic variances, we found evidence for a significant decrease of forehead patch size over time in older birds. This change appears to be caused by a change in the sign of viability selection on forehead patch size, which is associated with a decline in the breeding value of multiple breeders. Our data thus reveal complex patterns of environmental influence on the expression of secondary sexual characters, which may have important implications for understanding selection and evolution of these characters. PMID- 15119447 TI - Genetic basis of the trade-off between offspring number and quality in the bank vole. AB - One of the main tenets of modern life-history theory is the negative relationship (trade-off) between the number and quality of offspring produced. Theory predicts a negative genetic correlation between these traits since both are closely related to fitness of individuals. However, the genetic basis of the trade-off has only been tested to a limited extent in natural populations. We examined whether size and quality of offspring are negatively related to litter size in the bank vole Clethrionomys glareolus. First, we found a significant negative phenotypic correlation between the number and size of offspring at birth in both laboratory and field populations of the bank vole. Second, a larger size at birth decreased the maturation age of female offspring in the laboratory, and increased the probability of breeding and the size of the first litter in the field. Furthermore, manipulation of offspring size at weaning indicated that structural effects of birth size in mammals have a more profound effect on the expression of life-history traits than weaning size. Finally, in addition to the phenotypic negative correlation between the number and size of offspring, we found evidence for a negative genetic correlation between these two traits, which confirms the genetic basis of the trade-off. This negative genetic covariation may have considerable effects on the rate and direction of evolution of the two related life-historical traits. PMID- 15119448 TI - Sperm-limited fecundity in nematodes: how many sperm are enough? AB - The Bateman principle, which holds that oocytes are the limiting gamete in reproduction, is violated in a variety of species. Self-fertilizing hermaphrodites of the nematode Caenorhabditis elegans provide an example of a system in which sperm number limits lifetime reproductive output, in this species due to the protandrous nature of sperm production that in turn delays the onset of fertilization. This reproductive delay forms the basis of a trade-off between generation time and total fecundity, in which sperm number plays a pivotal role. I use an age-structured population model to describe the number of sperm that maximize fitness, given larval development time and rates of gamete production. The model predicts the evolution of sperm numbers that are consistent with empirical data for C. elegans provided that precocious larval sperm production is taken into account. Several testable hypotheses follow from the model regarding how natural selection and environmental variation may influence patterns of sperm production among populations or species with a similar mode of reproduction. PMID- 15119449 TI - Using comparative genomic data to test for fast-X evolution. AB - Genes may acquire nonsynonymous substitutions more rapidly when X-linked than when autosomal, but evidence for "fast-X evolution" has been elusive. Fast-X evolution could explain the disproportionate contribution of X-linked genes to hybrid sterility and other traits. Here, we use a comparative genomic approach, with sequences of 30-110 genes in four Drosophila species, to test for fast-X evolution. Specifically, the 3L autosome arm in D. melanogaster and D. simulans is homologous to the right arm of the X chromosome in D. pseudoobscura and D. miranda. We executed two paired comparisons to determine how often genes on this chromosome arm exhibit higher rates of nonsynonymous substitution in the D. pseudoobscura species group, as predicted by fast-X evolution. We found a statistically significant pattern consistent with fast-X evolution in one comparison and a similar trend in the other comparison. Variation in functional constraints across genes may have masked the signature of fast-X evolution in some previous studies, and we conclude paired comparisons are more powerful for examining rates of evolution of genes when X-linked over autosomal. PMID- 15119450 TI - Nonlinear growth cost in Menidia menidia: theory and empirical evidence. AB - Juvenile growth is submaximal in many species, suggesting that a trade-off with juvenile growth must exist. In support of this, recent studies have demonstrated that rapid growth early in life results in decreased physiological performance. Theory clearly shows that for submaximal growth in juveniles to be optimal, the cost of growth must be nonlinear. However, nearly all of the empirical evidence for costs of growth comes from linear comparisons between fast- and slow-growing groups. It is consequently unclear whether any known cost can account for the evolution of submaximal juvenile growth. To test whether the cost of growth exhibits the logically necessary nonlinearity, we measured critical swimming speed (Ucrit), the maximum speed sustained in incremental velocity trials, in Atlantic silversides, a species for which the costs and benefits of growth are well studied. To increase our ability to detect a nonlinear relationship between Ucrit, a proxy for juvenile fitness, and growth, we manipulated ration levels to produce a broad range of growth rates (0.16 mm/day(-1) to 1.20 mm/day(-1)). Controlling for size and age, we found that Ucrit decreased precipitously as growth approached the physiological maximum. Using Akaike's information criterion, we show that swimming performance decreases with the square of growth rate, providing the first demonstration of a nonlinear cost of growth. PMID- 15119451 TI - Cryptic female preference for colorful males in guppies. AB - Cryptic female choice (CFC) refers to female-mediated processes occurring during or after copulation that result in biased sperm use in favor of preferred or compatible males. Despite recent empirical support for this hypothesis, evidence that CFC contributes towards the evolution of male body ornaments, in the same way that precopulatory female choice does, is currently lacking. Here, we tested the possibility that CFC selects for increased male attractiveness in the guppy Poecilia reticulata, a freshwater fish exhibiting internal fertilization. Specifically, we examined whether females are able to manipulate the number of sperm transferred or retained at copulation in favor of relatively attractive males. In support of this prediction, we found that following solicited copulations the number of sperm inseminated is influenced exclusively by the female's perception of relative male coloration, independent of any direct manipulation of males themselves. Because females prefer brightly colored males during precopulatory mate choice, our finding that colorful males are also favored as a consequence of enhanced insemination success indicates that cryptic female choice can reinforce precopulatory preferences for extravagant male ornaments. PMID- 15119452 TI - Discerning between recurrent gene flow and recent divergence under a finite-site mutation model applied to North Atlantic and Mediterranean Sea fin whale (Balaenoptera physalus) populations. AB - Genetic divergence among conspecific subpopulations can be due to either low recurrent gene flow or recent divergence and no gene flow. Here we present a modification of an earlier method developed by Nielsen and Wakeley (2001), which accommodates a finite-site mutation model, to assess which of the two models of divergence is most likely given the observed data. We apply the method to nucleotide sequence data collected from the variable part of the mitochondrial control region in fin whales (Balaenoptera physalus) from the Atlantic coast off Spain and the Mediterranean Sea. Our estimations strongly favor a model of recurrent gene flow over a model of recent divergence and zero gene flow. We estimated the migration rate at two females per generation. While the estimated rate is high by evolutionary standards, exchange rates of this order of magnitude is low from an ecological and conservation perspective and entirely consistent with the current paucity of fin whale sightings in the Strait of Gibraltar today. Intensive commercial shore-based whaling during the 1920s removed substantial numbers of fin whales in the Strait of Gibraltar and this local population has seemingly since failed to recover. PMID- 15119453 TI - A lesson in successful implantation. PMID- 15119454 TI - Implant biomechanics in grafted sinus: a finite element analysis. AB - This in vitro study investigated the stress distribution in the bone surrounding an implant that is placed in a posterior edentulous maxilla with a sinus graft. The standard threaded implant and anatomy of the crestal cortical bone, cancellous bone, sinus floor cortical bone, and grafted bone were represented in the 3-dimensional finite element models. The thickness of the crestal cortical bone and stiffness of the graft were varied in the models to simulate different clinical scenarios, representing variation in the anatomy and graft quality. Axial and lateral loads were considered and the stresses developed in the supporting structures were analyzed. The finite element models showed different stress patterns associated with helical threads. The von Mises stress distribution indicated that stress was maximal around the top of the implant with varying intensities in both loading cases. The stress was highest in the cortical bone, lower in the grafted bone, and lowest in the cancellous bone. When the stiffness of the grafted bone approximated the cortical bone, axial loading resulted in stress reduction in all the native bone layers; however, lateral loading produced stress reduction in only the cancellous bone. When the stiffness of the graft was less than that of the cancellous bone, the graft assumed a lesser proportion of axial loads. Thus, it caused a concomitant stress increase in all the native bones, whereas this phenomenon was observed in only the cancellous bone with lateral loading. The crestal cortical bone, though receiving the highest intensity stresses, affected the overall stress distribution less than the grafted bone. The stress from the lateral load was up to 11 times higher than that of the axial load around the implant. These findings suggest that the type of loading affects the load distribution more than the variations in bone, and native bone is the primary supporting structure. PMID- 15119455 TI - Effect of enamel matrix derivative (Emdogain) on bone defects in rabbit tibias. AB - The aim of this study was to assess the effect of an enamel matrix derivative (Emdogain, Biora, AB, Malmo, Sweden) on bone healing. Ten New Zealand rabbits, weighing about 2.5 kg, were used. One 8-mm bone defect was created in each tibia. The defect on the right leg was filled with Emdogain, whereas the defect on the opposite leg was left unfilled as control. A total of 20 defects were created. Five rabbits each were killed at 4 and 8 weeks with an overdose of Tanax. Block sections containing the defects were retrieved and the specimens processed for light microscopy examination. The slides were stained with acid and basic fuchsin and toluidine blue. Histologically, no differences were noted in both groups at each observation period; in the test group, remnants of the implanted Emdogain were not present at 4 weeks. Newly formed bone was detectable in both groups at all observation times. At 8 weeks, both groups showed mature bone, and in the test group the material implanted was not visible. No inflammatory cells were visible in both groups. In conclusion, our results indicate that Emdogain implanted in bone defects is fully resorbed after 4 to 8 weeks and does not adversely affect bone formation. PMID- 15119456 TI - Human histologic analysis of mineralized bone allograft (Puros) placement before implant surgery. AB - Because clinicians are placing more dental implants, it is becoming more important to maintain bone volume after tooth extraction. This article discusses the various bone-augmentation materials available to the clinician and illustrates a case report of particulate mineralized bone allograft (Puros) placement after extraction. Exposure of the grafted site after 5 months revealed a hard bony structure. Human histologic analysis at the light microscopic level revealed nonvital spicules of mature calcified bone having a highly organized matrix surrounded by viable noncalcified immature bone matrix, or osteoid. It was concluded that mineralized human allograft demonstrated the formation or remodeling of bone histologically and was clinically useful to maintain bone volume for implant placement after extraction. To the authors' knowledge, this is the first publication to demonstrate human histology of particulate mineralized bone allograft (Puros) after placement into an extraction site. PMID- 15119457 TI - Technique for achieving a passive framework fit: a clinical case report. AB - The clinical application of a 4-step technique for achieving the passive fit of an implant-supported cast bar or framework is reported. Implant-level transfers were attached, splinted intraorally with pattern resin, picked up in an open-tray elastomeric impression, and used to fabricate a working cast containing implant analogs. A light-cured template was used to verify master cast accuracy. A ceramometal fixed partial denture restoration was fabricated on the working cast with a preliminary try-in of the cast metal framework. A passive prosthesis fit was achieved. Failure to create an accurate working cast can distort the intended fit of a cast framework on the abutments in the patient's mouth. Optimal positioning of the implant analog in the master cast depends on (1) the transfer technique and (2) the ability of the system to maintain precise rotational orientation of the transfer components. The splinted transfer technique developed 20 years ago has sometimes been effective in improving the accuracy of framework fit with external hexagon implant-abutment connections, but it has not been documented with newer implant-abutment connections. Techniques for reestablishing master cast accuracy and correcting the superstructure are presented. The use of the presented technique resulted in a passive-fitting framework. PMID- 15119458 TI - Oral triazolam sedation in implant dentistry. AB - Triazolam can be helpful for sedating dental implant patients when administered orally or sublingually in low dosages of 0.125 or 0.25 mg, but not exceeding 0.5 mg. It is a fast- but short-acting benzodiazepine with few side effects, and it has a long record of successful use. Its effects can be reversed with incremental intravenous flumazenil, although there is a risk of seizure. Triazolam has not been shown to be carcinogenic, and it has a low potential for abuse and addiction. It is contraindicated in patients who are pregnant, breast-feeding, and those concomitantly taking ethanol, macrolid antibiotics, some protease inhibitors, psychotropic medications, ketoconazole, itraconazole, nefaxodone, or other medications that impair oxidative metabolism mediated by cytochrome P450 3A (CYP 3A). Triazolam should be used with caution in patients taking grapefruit juice, cyclosporine, and other drugs such as calcium channel blockers including nifedipine, verapamil, and diltiazem. The lowest effective dose should be used. PMID- 15119459 TI - Measuring outcome measures. PMID- 15119460 TI - Human frequency-following response correlates of the distortion product at 2F1 F2. AB - We characterized the 2F1-F2 distortion product reflected in the human frequency following response (FFR). In the first experiment, we evaluated the input-output growth functions of the distortion product at 2F1-F2 (FFR-DP) for three primary pairs. In the second experiment, we tested the effect of primary tone level variation on the FFR-DP. The results for all three stimulus pairs showed that while the amplitude of FFR-DP increased with stimulus intensity, the slope of the amplitude growth decreased with increasing frequency. Consistent with distortion product otoacoustic emission (DPOAE) data, our observations suggest that there is a distinct region where the separation of the primary tone levels produces maximal distortion. The robust FFR-DP measure could complement the less reliable DPOAE at low frequencies and when middle ear pathology precludes its measurement. PMID- 15119461 TI - Clinical experience with the vestibular evoked myogenic potential. AB - The vestibular evoked myogenic potential (VEMP) is a promising test of the descending vestibulocollic system. Our aim was to determine whether the VEMP can be applied to an older patient population and can detect lesions in descending vestibulospinal pathways. We also compared VEMP clinical performance with that of the standard caloric test. VEMP test performance was retrospectively analyzed in relation to clinical diagnosis and other vestibular test performance in 62 patients (age, 30-85 years) referred for vestibular testing to Mayo Clinic, Jacksonville, Florida. The VEMP was evoked using a 250 Hz tone burst. Results suggest age-related changes in VEMP amplitude and latency in this patient population. VEMP tests were sensitive to lesions not detected by electronystagmography. VEMP and caloric sensitivity and specificity were essentially equal (d' = 1). Combining both tests improved sensitivity. However, VEMP false-positive rates hampered specificity. VEMP testing may be refined to improve false-positive rates and clinical utility. PMID- 15119462 TI - Hearing-in-Noise: comparison of listeners with normal and (aided) impaired hearing. AB - In this study, the performance of 48 listeners with normal hearing was compared to the performance of 46 listeners with documented hearing loss. Various conditions of directional and omnidirectional hearing aid use were studied. The results indicated that when the noise around a listener was stationary, a first- or second-order directional microphone allowed a group of hearing-impaired listeners with mild-to-moderate, bilateral, sensorineural hearing loss to perform similarly to normal hearing listeners on a speech-in-noise task (i.e., they required the same signal-to-noise ratio to achieve 50% understanding). When the noise source was moving around the listener, only the second-order (three microphone) system set to an adaptive directional response (where the polar pattern changes due to the change in noise location) allowed a group of hearing impaired individuals with mild-to-moderate sensorineural hearing loss to perform similarly to young, normal-hearing individuals. PMID- 15119463 TI - Speech evoked cortical potentials: effects of age and stimulus presentation rate. AB - We examined the effects of stimulus complexity and stimulus presentation rate in ten younger and ten older normal-hearing adults. A 1 kHz tone burst as well as a speech syllable were used to elicit the N1 -P2 complex. Three different interstimulus intervals (ISI) were used (510, 910, and 1510 msec). When stimuli were presented at the medium presentation rate (910 msec ISI), N1 and P2 latencies were prolonged for older listeners in response to the speech stimulus but not the tone stimulus. These age effects were absent when stimuli were presented at a slower rate (1510 msec ISI). Results from this study suggest that rapidly occurring stimulus onsets, either within a stimulus or between stimuli, result in prolonged N1 and P2 responses in older adults. This is especially true when processing complex stimuli such as speech. One potential explanation for this age effect might be age-related refractory differences in younger and older auditory systems. Refractory issues might in turn affect synchronized neural activity underlying the perception of critical time-varying speech cues and may partially explain some of the difficulties older people experience understanding speech. PMID- 15119464 TI - Hearing specific and generic measures of the psychosocial impact of hearing aids. AB - Hearing-specific and generic measures of hearing aid outcome were examined in order (a) to determine their relative sensitivity to hearing aid use and (b) to examine the relationship between pre-hearing aid use expectations and post-use outcomes. Ninety-two hearing-impaired individuals completed some combination of the Abbreviated Profile of Hearing Aid Benefit, Expected Consequences of Hearing Aid Ownership (ECHO), Satisfaction with Amplification in Daily Life (SADL), and Psychosocial Impact of Assistive Devices Scale, and provided reports of their daily and lifetime hearing aid use. In general, (a) the longer individuals wear hearing aids, the more positive the reported outcome, and (b) ECHO scores of non hearing aid users are higher than SADL scores of new hearing aid users (six weeks to one year of use) but are similar to those obtained from experienced users (greater than one year of use). Between-questionnaire comparisons showed the generic measure to be as sensitive as the hearing aid specific measures. We suggest that generic measures have some advantages over hearing specific measures but that each has a place in the clinic. PMID- 15119465 TI - Gain and maximum output of two electromagnetic middle ear implants: are real ear measurements helpful? AB - We compared the output of two electronic middle ear implants: the Otologics MET device and the Vibrant Soundbridge device. Both devices were programmed in the linear amplification mode. Aided minus unaided sound pressure levels recorded in the ear canal (objective gain) were compared to unaided minus aided soundfield thresholds (functional gain) in 13 patients with severe sensorineural hearing loss. In addition, input/output characteristics were studied with the help of ear canal measurements. Objective gain was consistently lower than functional gain, with wide variation between patients and frequencies. Using input/output data measured in the ear canal in combination with functional gain data, the mean maximum output of the two devices was estimated, expressed in dB SPL. In comparison to NAL-R target values, (functional) gain was adequate; however, the maximum output was low, especially for the Vibrant Soundbridge device. PMID- 15119466 TI - Detection of ophthalmic impairments indirectly with electronystagmography. AB - The objective was to develop, from a pool of clinical electronystagmography (ENG) data, normal lower limits for the corneo-retinal potential (CRP). The CRP evaluated in the present study was derived as a byproduct of eye movement calibration with a computerized ENG system. The data set was collected from a cohort of patients without history of ophthalmic disease. This normative study was designed to develop upper and lower limits for the CRP recorded indirectly from ENG testing. Subjects were 107 consecutive patients (41 males, mean age 57 years). Subject age did not, but gender did affect significant changes in the CRP. Specifically women showed larger CRP values than men. Case studies are presented that support the contention that the dark-adapted CRP may be helpful in the identification of patients with ophthalmic diseases known to affect the CRP and, thus, augment information normally obtained in the course of the ENG examination. PMID- 15119467 TI - Egyptian Smoking Prevention Research Institue (ESPRI). AB - The overall goal of the ESPRI is to reduce tobacco use in Egypt by carrying out interlocking observational, intervention and policy research, and through capacity building for research, prevention, communications and policy on smoking prevention. Having the ability to rely on ESPRI for the best scientific knowledge will add momentum to the current efforts underway to curb smoking in Egypt. PMID- 15119468 TI - Nicotine dependance among adult male smokers in rural Egypt. AB - Nicotine dependence is a significant public health problem. This study describes the nicotine dependence status among male adults in rural communities in Egypt. A survey was carried out in five rural villages in Egypt to study the smoking prevalence. A total of 938 current smokers were identified and their nicotine dependence status was studied. About 9% of all smokers in the studied villages were found to have heavy dependence to nicotine. Heavy dependence was associated with younger age of smoking initiation (p<0.05) and more smoking in the first hours of the day (p<0.001). Heavy dependent smokers are less likely to quit smoking (p<0.001), lack the confidence to quit by themselves (p<0.001) and less likely to have tried to quit earlier (p<0.001). Dependent smokers are more likely to smoke in the presence of their children (p<0.001). Reasons for smoking included the habit of smoking helping them to keep them going when tired, to make them alert and not knowing what to do with their hands without a cigarette. The main reasons they identified for restarting smoking after quitting were the signs of withdrawal namely headaches, irritability and difficulty in concentration. Nicotine dependence status and attributes were comparable to studies reported in other countries around the world. Enhanced behavioral and medical intervention strategies are needed to motivate helping both low and heavy nicotine dependent smokers to increase the number and effectiveness of quit attempts. PMID- 15119469 TI - Prevalence of smoking among rural secondary school students in Qualyobia governorate. AB - A cross sectional study was carried out in two rural secondary schools in Qualyobia governorate in the academic year 2002-2003 in order to assess the students' knowledge, attitudes and practice toward smoking. An anonymous self administered questionnaire was used. A total of 635 students, 416 males and 219 females were interviewed. The response rate was 100%. The prevalence of ever tried cigarette was 29% (40% among males and 7% among females) with a median age of initiation at 11 years of age. The prevalence of current cigarette smoking among all students was 7.7 (11.5% among males and 0% among females). Waterpipe (Shisha) smoking was reported by 19% (26% among males and 5% among females) with median age of initiation at 12 yearsof age. The risk of smoking is higher among sons of highly educated mothers (OR=3.7, 95% CI=1-12), Boy only school students have almost half the risk of smoking than mixed school male students (OR=0.4, 95% CI=0.2-0.8). On studying the effect of having a smoker family member or friends on students smoking, having a smoker friend increases the risk of male students three time (OR=3, 95%CI=2-5) while others smoking was not statistically significant. More than 90% of the students (both males and females) thought that active and passive smoking are harmful. More females than males thought smoking is addictive (89% of the females and 81% of the males) while more males than females agreed that smokers have more friends (34% of males, 26% of the females) and that smoking makes boys look handsome (8% of males, 3% of females). PMID- 15119470 TI - Review on water pipe smoking. AB - Water-pipe smoking may lead to cancers, genetic damage, diseases of the lungs and other disease conditions. Many of the studies on these subjects are merely anecdotal or lack the necessary rigorous study design or the power needed to be certain of the results. Given the large number of people who smoke waterpipe and the fact that waterpipe smoking has become a fashionable trend in the Middle East and the Western world among the youth, it is essential to study the health effects of waterpipe smoking with renewed emphasis. PMID- 15119471 TI - Water pipe (Sisha) smoking in cafes in Egypt. AB - Shisha cafe patrons in Cairo, Egypt were interviewed to assess their knowledge, attitudes and practices regarding smoking and health. The median age of initiation of Shisha smoking is 20 years. Shisha smokers know about the hazards of smoking and believe that Shisha smoking is less dangerous than cigarette smoking. Over half the Shisha smokers have tried to quit in the past year. The younger adults who smoke Shisha also tend to smoke more often with friends, smoke cigarettes in addition to Shisha and prefer fruit flavored tobacco as compared to tobacco mixed with molasses favored by Shisha smokers who are older. Heavy Shisha smoking was not related to age. PMID- 15119472 TI - Impact of religious rulings (Fatwa) on smoking. AB - An interview survey was carried out in a rural village and two nearby schools in Qalyubia Governorate to assess the pattern of smoking and knowledge about religious ruling (Fatwa) and its impact on the quit attempts. Also, a similar survey was conducted in 6 Shisha cafes in Cairo. The results showed that the majority of respondents (81% among rural adults, 83.2% among Shisha cafe patrons, 73.3% among rural youth and 81.4% among rural students) knew about the Fatwa on smoking. Higher proportions of all participants thought that smoking is a sin (97.3% among rural adults, 80.8% among Shisha cafe patrons, 94.4% among rural youth and 98.4% among rural students). There was a significantly higher knowledge about Fatwa on smoking among men than women. This indicates a successful outreach program targeted mainly to men through mosques. Knowledge about Fatwa on smoking increased significantly with increased exposure to antismoking messages from religious leader. Knowledge about the Fatwa on smoking or belief that smoking is a sin had no significant effect on quit attempts. Our results point to the need for intensive efforts on the part of religious leaders to translate the current belief that smoking is a sin into quitting among smokers. Better results may be achieved through personal interactions in small groups rather than in mosque settings. PMID- 15119473 TI - University advocacy on tobacco prevention in Egypt: partnership between Ain Shams University and the Egyptian Smoking Prevention Research Institute (ESPRI). PMID- 15119474 TI - Smoking prevention and treatment in Egypt. AB - The basic ingredients for a successful prevention program for Egypt are all present but to be successful, these efforts need to be maintained year round with additional resources for prevention and treatment made available to reach out to all regions and neighborhoods in Egypt. We need to expand our treatment capabilities by training primary care physicians to be more active in the area of smoking prevention and treatment. PMID- 15119475 TI - Pharmacologic treatment of autism. AB - Autism is a chronic and lifelong pervasive developmental disorder for which there is yet no effective cure, and medical management remains a major challenge for clinicians. In spite of the possible similarities with conditions that have an established pharmacotherapy, and despite improvements in some associated "problematic behaviors" following the use of available medications, effective medical treatment for the core symptoms involving language and social cognition remains elusive. The purpose of the present article is to review current biologic knowledge about autism in an attempt to correlate clinical trials with known mechanisms of disease. In addition, the need for controlled studies and for the creation of homogeneous subgroups of patients based on clinical and genetic characteristics is emphasized. The application of molecular genetic investigations and pharmacogenetics in the diagnostic work-up of autistic patients can lead to more effective individualized medical care. PMID- 15119476 TI - Treating autistic spectrum disorders in children: utility of the cholinesterase inhibitor rivastigmine tartrate. AB - Rivastigmine tartrate is a dual-action cholinesterase inhibitor shown to improve language, cognition, and global functioning in patients with Alzheimer's disease, likely via increased availability of cerebral acetylcholine. Because cholinergic receptor abnormalities can contribute to the neuropathology of autistic spectrum disorders, rivastigmine tartrate could prove to be an effective therapy for affected children. Observations of improved behavior and language output from prior open-label and double-blind treatment of autistic children with donepezil, another cholinesterase inhibitor, prompted this 12-week open-label study with rivastigmine tartrate of 32 autistic patients. Therapeutic indices were the Childhood Autistic Rating Scale, Gardner's Expressive and Receptive One-Word Picture Vocabulary tests, and the Conners' Parent Rating Scale. Testing administered at baseline, 6 weeks, and 12 weeks showed gains in both expressive speech and overall autistic behavior over baseline. These improvements were statistically significant and supported the hypothesis that treatment with cholinergic enhancing drugs in autistic spectrum disorders yields positive therapeutic effects. PMID- 15119477 TI - Primary generalized epilepsy during infancy and early childhood. AB - The present study delineates a benign generalized epileptic disorder during infancy and early childhood similar to the well-defined syndrome of primary generalized epilepsy in adolescence. The inclusion criteria for the study required infants under the age of 4 years mainly presenting with generalized nonfebrile seizures, requiring corroboration with generalized epileptic discharges on electroencephalograms (EEGs); an unremarkable pregnancy, labor, and perinatal course; a favorable response to antiepilepsy drugs, preferably monotherapy; and a normal cognitive outcome. The clinical features of seizures, EEG correlates, response to medications, developmental outcome, and family history were analyzed. Twenty-five infants fulfilled the inclusion criteria, presenting at ages 4 to 36 months (mean 17 months) with recurrent generalized clonic seizures, which were commonly short-lived, lasting up to 5 minutes; two infants also had status epilepticus. Fourteen infants (56%) had accompanying febrile seizures, which preceded the nonfebrile seizures in 10 of them. A positive family history of seizures was found in 8 (32%) patients. Analysis of the EEG showed generalized epileptiform discharges in the form of 3 to 4 Hz spike wave and normal background activity in 21 patients (84%), with a photosensitive response induced in 3 children. A larger group of 18 infants promptly responded to therapy, mainly valproic acid, which was terminated after 2 years, along with EEG normalization and no recurrence of seizures. A smaller group of 7 patients require prolonged therapy that keeps them seizure free; the EEG remains paroxysmal, and the seizures could recur when treatment is discontinued. All patients are presently seizure free within a follow-up period of 1.5 to 14 years. Their cognition is normal, but 12 patients have short attention and concentration spans, impulsiveness, and learning difficulties. As such, the data presented here delineate an idiopathic generalized epileptic disorder during infancy with a benign course, a rapid response to therapy, and preservation of cognitive skills that may be added to the current classification of the epileptic syndromes. PMID- 15119478 TI - Congenital myasthenic syndrome: presentation, electrodiagnosis, and muscle biopsy. AB - We report 10 children with congenital myasthenic syndromes diagnosed by clinical features, electrodiagnostic studies, and response to acetylcholinesterase inhibitors. Age at diagnosis (mean = 4.4 years; range 0.2-10 years) correlated with age fatigue was recognized. Symptoms at presentation included mild gross motor development delay (7/10), speech articulation difficulty (5/10), and respiratory and feeding difficulties resulting in poor growth in 7 of 10 children. None of the five children with possible presynaptic abnormalities had decremental compound muscle action potential responses to 2 Hz repetitive nerve stimulation. Instead, electrodiagnostic studies showed a more than 100% increment of compound muscle action potential amplitude during 50 Hz repetitive nerve stimulation in two children and sustained compound muscle action potential decrement to 2 Hz repetitive nerve stimulation after depletion (10 Hz stimulation for 10 min) in four children. Muscle biopsies (n = 7) showed mild to severe variation in fiber size. Our experience suggests that many children with congenital myasthenic syndromes might be undiagnosed because of atypical presentation and because additional electrophysiologic studies are required. PMID- 15119479 TI - Risk analyses for the cognitive phenotype in Turner's syndrome: evidence of familial influence as a decisive factor. AB - The interindividual varying cognitive performance in female patients with Turner's syndrome has usually been attributed to the interindividual varying mosaicism with a consecutive variable loss of X-chromosome DNA or to secondary risk factors such as estrogen deficiency owing to ovarian failure. The aim of our study was to determine the specific impact of X chromosome-related features and associated risk factors, on the one hand and familial influences, on the other hand on the interindividual variation in the cognitive phenotype. One hundred and one subjects with Turner's syndrome and 53 sisters as controls for familial influences were examined by comparing the cognitive information processing abilities (Kaufmann Assessment Battery for Children [K-ABC]). Subjects with Turner's syndrome performed at a significantly lower level than sisters on all subscales (eg, Mental Processing Composite: Turner's syndrome 86.4 [SD 15.0] versus sisters 99.3 [SD 10.6]; P < .001). For the neurocognitive phenotype in subjects with Turner's syndrome, a significant correlation was found only with the sisters' cognitive abilities (Mental Processing Composite: r = .38, P < .05). In contrast, neither the individual mosaic status nor the known associated risk factors predicted the neurocognitive phenotype in Turner's syndrome. These results are corroborated in the regression analyses in those subjects with Turner's syndrome with a sister (Simultaneous Processing(sister) for Simultaneous Processing(Turner's syndrome): beta = .346, P < .05, corrected R2 = .049; and Mental Processing Composite(sister) for Mental Processing Composite(Turner's syndrome): beta = .354, P < .05, corrected R2 = .033). The interindividual variation of intellectual abilities in Turner's syndrome seems to be primarily related to familial coinfluences and not to the interindividual varying loss of X chromosome DNA in terms of hidden mosaicism or potential associated risk factors. PMID- 15119480 TI - Guillain-Barre syndrome in children. AB - In industrialized nations with widespread immunization programs, Guillain-Barre syndrome is the most common cause of acute paralytic illness in children and adults. The incidence of the disease has been estimated to range from 0.5 to 1.5 in 100,000 in individuals less than 18 years of age. Approximately 15% of children with Guillain-Barre syndrome develop respiratory failure and require mechanical ventilatory support. Prospective randomized treatment trials in childhood Guillain-Barre syndrome are wanting; however, smaller case series studies using historical controls suggest that both plasmapheresis and administration of human immunoglobulin could be helpful in reducing morbidity in children with Guillain-Barre syndrome. The prognosis for recovery in children is generally excellent, with the majority of children achieving a complete functional recovery within 6 months from the onset of illness. Studies using an animal model of human Guillain-Barre syndrome, experimental allergic neuritis, have expanded our understanding of the pathogenesis of the disease and suggest new directions for exploration in the treatment of this disorder. PMID- 15119481 TI - Congenital club foot with survival of motor neuron 1, telomeric (SMN1) gene deletion. AB - A boy with nonreducible bilateral congenital talipes equinovarus had delayed milestones with early-onset generalized hypotonia and muscular weakness. The condition remained stable until he was 8 years old. A slow worsening of motor abilities, with myopathic signs, was observed thereafter. A homozygous deletion of exons 7 and 8 of the survival of motor neuron 1, telomeric (SMN1) gene was found, without neuronal apoptosis inhibitory protein (NAIP) gene deletion, leading to the diagnosis of spinal muscular atrophy. Independent ambulation was lost when he was 13 years old. The occurrence of congenital clubfoot with early onset of neurologic signs, but with a very slowly progressive course, has not been reported in spinal muscular atrophy until now. PMID- 15119482 TI - Cerebral and cerebellar motor activation abnormalities in a subject with Joubert syndrome: functional magnetic resonance imaging (MRI) study. AB - Joubert syndrome is an autosomal recessive disorder characterized by hypotonia, ataxia, developmental delay, and a distinctive hindbrain malformation involving the cerebellum and brain stem, visualized radiographically on magnetic resonance imaging (MRI) as the "molar tooth sign." In postmortem brains from subjects with Joubert syndrome, there is an apparent absence of decussation of both corticospinal and superior cerebellar tracts, although the functional significance has not been elucidated. We sought to explore the cerebral and cerebellar activation pattern elicited by finger tapping in an adolescent with Joubert syndrome and in a normal control subject using functional MRI. In contrast to the typical highly lateralized activation seen in our control subject, the subject with Joubert syndrome demonstrated striking bilateral activation of the sensorimotor and cerebellar cortex. Although our functional MRI data do not indicate a clear absence of decussation, the abnormal activation pattern observed suggests altered brain functional organization in relation to anatomic differences. Malformation of the hindbrain could result in recruitment of alternative pathways, similar to what has been observed following ischemic injury to the developing or mature central nervous system. PMID- 15119483 TI - Thrombophilia interpretation in childhood stroke: a cautionary tale. AB - Several authors have reported a link between childhood stroke and inherited thrombophilia in recent years. The impact of such a relationship on management and outcome is yet to be determined, as is the potential cost-benefit ratio associated with the performance of thrombophilic screening in children presenting with ischemic stroke. We present a case that highlights the need for clinical and radiologic examinations to remain the definitive criteria used to diagnose stroke in children. The diagnosis should not be influenced by the finding of a thrombophilic marker. PMID- 15119484 TI - Effects of medications on regulation of body temperature of patients with Tourette syndrome. AB - Tourette syndrome is defined and characterized mainly by the presence of motor and phonic tics. Frequently, other medical, psychologic, and psychiatric symptoms coincide with Tourette syndrome. Despite extensive efforts extended over many years of research, the etiology leading to Tourette syndrome remains obscure. A number of hypotheses have been offered in the past to resolve the pathophysiology of Tourette syndrome. Based on the existence of an abnormal body temperature profile in patients with Tourette syndrome, an idiopathic hypothalamic disorder has been proposed to be involved in this ailment. When monitoring the effects of medications employed in the consensus for Tourette syndrome treatments, it became evident that medications ameliorate neurologic tics and rectify the patient's periodic hypothermal body temperature. Correlating the neurologic status to hypothalamic dysregulation, the data presented in this study strengthen the hypothesis of an idiopathic hypothalamic disorder underlying the Tourette syndrome cascade. PMID- 15119485 TI - Acute disseminated encephalomyelitis following pleural empyema owing to Boerhaave's syndrome. AB - Acute disseminated encephalomyelitis is reported in a 14-year-old male patient associated with a pleural empyema caused by an esophageal perforation. During the hospital course, the patient experienced an episode of cardiac arrest, raising the question of hypoxic-ischemic cerebral injury, but the magnetic resonance imaging (MRI) findings and clinical course were most consistent with acute disseminated encephalomyelitis. After a course of methylprednisolone, broad spectrum antibiotics, and bilateral chest tube placement, the patient recovered with only mild residual deficits, along with resolution of his MRI findings. PMID- 15119486 TI - Neuroepithelial cysts in a patient with Joubert syndrome plus renal cysts. AB - Joubert syndrome is a rare genetic neurologic disorder associated with hypoplasia or absence of the cerebellar vermis. The classic form is characterized by ataxia, hypotonia, eye movement abnormalities, developmental delay, and abnormal breathing patterns. In contrast, other patients have the additional feature of kidney cysts. This population could represent a distinct form of Joubert syndrome. One case of Joubert syndrome with subcortical neuroepithelial cysts was recently described. We report a new case of Joubert syndrome with overlapping features, including diffuse progressive central nervous system neuroepithelial cysts and kidney cysts. Our data suggest that neuroepithelial cysts occur in conjunction with Joubert syndrome associated with kidney cysts. PMID- 15119487 TI - Naegleria fowleri hemorrhagic meningoencephalitis: report of two fatalities in children. AB - Two cases of hemorrhagic meningoencephalitis secondary to Naegleria fowleri infection confirmed by postmortem analysis are described. The first patient is a 5-year-old boy who presented with a severe headache, neck stiffness, and lethargy. His neurologic examination was significant for somnolence and nuchal rigidity. Cerebrospinal fluid studies and structural neuroimaging were consistent with hemorrhagic meningoencephalitis. Another 5-year-old boy presented to a different institution 2 miles away in the same week with similar complaints. Both patients declined rapidly and expired within 48 hours of admission secondary to transtentorial herniation caused by the mass effect of inflammation, edema, and hemorrhage with displacement of the brain stem. Histopathologic and immunochemistry analysis of brain tissue revealed the presence of Naegleria trophozoites in both cases. PMID- 15119489 TI - Legislature's actions improve state's healthcare environment; primary election critical. PMID- 15119488 TI - Subacute sclerosing panencephalitis with fulminating course: follow-up magnetic resonance spectroscopy (MRS) findings. AB - The fulminating form of subacute sclerosing panencephalitis is an extremely rare condition. Imaging findings are usually not correlated with clinical staging. We describe a 4-year-old girl with severe neuronal loss, demyelination, and gliosis in subcortical white matter by magnetic resonance spectroscopic examination even though she was diagnosed as clinical stage II. In 2 months' time, her clinical status worsened significantly. Follow-up magnetic resonance spectroscopy revealed findings that were consistent with clinical status. It is our opinion that magnetic resonance spectroscopy could demonstrate a rapidly progressive fulminating course of subacute sclerosing panencephalitis even in the early clinical stages. PMID- 15119490 TI - A short history of pediatric orthopedics in West Virginia. PMID- 15119491 TI - Laparoscopic treatment of Morgagni-Larrey hernia. AB - This report describes the cases of two patients who underwent laparoscopic repair for treatment of Morgagni-Larrey hernia at United Hospital Center in Clarksburg. The first patient was a 40-year-old woman complaining of epigastric discomfort and tenderness. Her chest X-ray revealed an anterior cardiophrenic mass, and a CT scan showed a characteristic Morgagni hernia with incarcerated colon and omentum. After reduction of the incarcerated bowel and omentum, autosuture repair was carried out laparoscopically In the second case, a 22-year-old man with severe GERD was found to have a large Morgagni hernia with incarcerated transverse colon and omentum, which was discovered while he was undergoing an elective laparoscopic Nissen fundoplication. Laparoscopic repair of Morgagni hernia was carried out, in addition to Nissen fundoplication. Both patients did very well postoperatively. Minimally invasive laparoscopic repair can be successfully carried out, and mesh implantation is performed only in some cases. In order to avoid pleural injury, we prefer not to remove the hernia sac before a Morgagni hernia is repaired. Laparoscopic repair should be considered the standard and safest procedure for the treatment of a Morgagni hernia. PMID- 15119492 TI - Rectal function following permanent prostate brachytherapy. AB - The purpose of this study was to review the clinical presentation, dosimetry, quality of life assessment, and management of proctitis following permanent prostate brachytherapy. The permanent prostate brachytherapy literature was reviewed using MEDLINE searches to ensure completeness. Rectal complications primarily consist of mild, self-limited proctitis. Rectal bleeding has been correlated with rectal dose. Dose response analysis has demonstrated that the risk of proctitis strongly correlates with the volume of rectal tissue exposed to a given dose. The reported incidence of rectal ulceration/fistula formation is < or = 1%. Constipation significantly increases the radiation dose to the rectum and potentially may contribute to brachytherapy-related morbidity. Quality of life assessments obtained via patient-administered instruments and by physician interview have illustrated that long-term bowel dysfunction is relatively uncommon. Significant bowel dysfunction following prostate brachytherapy is relatively rare, but rectal bleeding has been correlated with radiation dose. Routine post-brachytherapy biopsies of the anterior rectal wall are strongly contraindicated. The continued elucidation of the etiology of bowel dysfunction should result in refinements in implant technique and decreased brachytherapy related morbidity. PMID- 15119493 TI - Utilizing CIWA-Ar to assess use of benzodiazepines in patients vulnerable to alcohol withdrawal syndrome. AB - To determine whether use of the revised Clinical Institute Withdrawal Assessment (CIWA-Ar) would better guide treatment for the Alcohol Withdrawal Syndrome (AWS), we prospectively studied 16 patients identified as alcohol dependent or with a positive blood alcohol level on admission. All patients were administered the CIWA-Ar. If it was > or = 10, the patient was randomized to a benzodiazepine. If the CIWA-Ar was < 10, the patient was observed and the CIWA-Ar was administered every eight hours for 48 hours. Of the 35 patients screened, 16 were enrolled. Seven patients had a score of > or = 10 and entered a benzodiazepine treatment program. The mean CIWA-Ar score was 18 +/- 10. The remaining nine patients had an initial CIWA-Ar < 10, with a mean score of 3.8 +/- 2.4. We safely withheld detoxification regimens in 9 of 16 patients based on CIWA-Ar scores. The CIWA-Ar may obviate over-utilization of benzodiazepines in patients with AWS. PMID- 15119494 TI - Keratosis pilaris in pregnancy: an unrecognized dematosis of pregnancy? AB - Keratosis pilaris (KP) is a common hyperkeratotic condition that most commonly presents as skin-colored follicular papules surrounded by erythema. These lesions often appear grouped together on the lateral aspect of the arms, thighs, and buttocks. Although the etiology of KP is uncertain, it is thought to be the result of the formation of an orthokeratotic plug, which blocks and dilates the orifice and upper portion of the follicular infundibulum (Figure 1). Histologically, mild perivascular mononuclear cell infiltrates are usually present in the adjacent dermis. Studies have indicated a role for hormonal influences in the development of KP. The five cases presented in this study demonstrate keratosis pilaris as a condition in which the onset or severity of the dermatosis may be linked to the hormonal changes of pregnancy. PMID- 15119495 TI - Legislature passes many key healthcare bills. PMID- 15119496 TI - Confidentiality important in smaller locations. PMID- 15119497 TI - Atherosclerosis, oxidative stress and glutathione peroxidase-1: a new kid on the block. PMID- 15119498 TI - Intracardiac echocardiography. Do we need a new ultrasonographic window? AB - Intracardiac echocardiography constitutes a new ultrasonographic window for the examination of the cardiovascular system, allowing a view of the heart and great vessels through a transvenous approach. Its use in interventional procedures, such as closure of secundum atrial septal defect and patent foramen ovale, transseptal puncture and radiofrequency ablation for arrhythmias is increasing sharply. The intracardiac echocardiography systems available today are of two types: mechanical and electronic scanning systems. Both enhance the visualization of the structures and of the endocardial surface inside the cardiac chamber where they reside, allowing the exclusive possibility of interaction between interventional instruments and the anatomic structures subjected to percutaneous treatment. The evolution of this methodology for the diagnosis and treatment of cardiac pathology requires the acquisition of new knowledge to ensure a correct and efficient use. The aim of this manuscript was to provide the clinical cardiologist with a complete overview on this topic. PMID- 15119499 TI - The care of adult patients with congenital heart defects: a new challenge. AB - The aim of the present article was to point out some of the more common challenges and needs for adult patients with congenital heart defects. We may reasonably calculate a population approximating 80,000 to 100,000 patients in Italy. The profile of this patient population will change over the next few decades. Not all congenital heart defects require the same level of expertise; for this reason an integrated national service is required. Ideally, specialist units should be established in appropriate geographic areas; complex patients need to be grouped according to the expertise, experience and management they require. Less specialized regional centers and outpatient clinics in interconnected districts with GUCH units should be created. Specialist units should accept responsibility for educating the profession, training the specialists, and sharing particular skills between each other. The debate on this subject is far from over; we wish to contribute to and stimulate the discussion. PMID- 15119500 TI - One-minute heart rate recovery after cycloergometer exercise testing as a predictor of mortality in a large cohort of exercise test candidates: substantial differences with the treadmill-derived parameter. AB - BACKGROUND: Previous work showed a strong inverse association between 1-min heart rate recovery (HRR) after exercising on a treadmill and all-cause mortality. The aim of this study was to determine whether the results could be replicated in a wide population of real-world exercise ECG candidates in our center, using a standard bicycle exercise test. METHODS: Between 1991 and 1997, 1420 consecutive patients underwent ECG exercise testing performed according to our standard cycloergometer protocol. Three pre-specified cut-point values of 1-min HRR, derived from previous studies in the medical literature, were tested to see whether they could identify a higher-risk group for all-cause mortality; furthermore, we tested the possible association between 1-min HRR as a continuous variable and mortality using logistic regression. RESULTS: Both methods showed a lack of a statistically significant association between 1-min HRR and all-cause mortality. A weak trend toward an inverse association, although not statistically significant, could not be excluded. CONCLUSIONS: We could not validate the clear cut results from some previous studies performed using the treadmill exercise test. The results in our study may only "not exclude" a mild inverse association between 1-min HRR measured after cycloergometer exercise testing and all-cause mortality. The 1-min HRR measured after cycloergometer exercise testing was not clinically useful as a prognostic marker. PMID- 15119501 TI - Clinical relevance of homocysteine levels in patients receiving coronary stenting for unstable angina. AB - BACKGROUND: We prospectively investigated whether plasma homocysteine (HCY) concentrations are related to target lesion revascularization (TLR) rates in patients with unstable angina undergoing stenting. METHODS: We enrolled 196 consecutive patients with at least one successful coronary stent implantation for unstable angina. RESULTS: The mean vessel diameter was 3.1 +/- 0.5 mm. At follow up (17.8 +/- 7.5 months), patients with higher HCY levels (> 17 micromol/l, 4th quartile) had similar TLR rates to the rest of the sample (11.1 vs 13.2%, p = 0.90). On the other hand, high HCY levels did seem to be associated with higher total (13.3 vs 0.7%, p = 0.001) and cardiac (6.7 vs 0%, p = 0.01) mortality rates. At multivariate analysis, only target vessel diameter independently predicted TLR, while both HCY levels and target vessel size predicted late total mortality. CONCLUSIONS: At least in patients with a mean vessel diameter > 3 mm, HCY levels cannot be taken as a prognostic indicator of in-stent restenosis for patients with unstable angina. However, in spite of successful percutaneous revascularization, HCY values do seem to strongly influence late mortality. PMID- 15119502 TI - Off-pump coronary surgery improves in-hospital and early outcomes in octogenarians. AB - BACKGROUND: A retrospective study on octogenarians who underwent off-pump (OPCAB) or conventional (CCAB) coronary artery bypass surgery undertaken to evaluate the in-hospital and early outcomes in terms of survival and cardiac and neurological events. METHODS: The design of the study was single-institutional, retrospective and comparative. Between January 1997 and May 2003, 114 patients were included and 73 underwent OPCAB, while 41 underwent CCAB. Uni- and multivariate analyses were used to determine the correlation between the pre- and intraoperative data and hospital death or complications. The overall survival and freedom from cardiac events were determined using Kaplan-Meier analysis. The linearized rates of follow-up complications were compared between the two groups. RESULTS: Overall, comparison of CCAB to OPCAB revealed differences in the operative mortality (6 deaths for CCAB, 14.6% vs 5 deaths for OPCAB, 6.8%; p = 0.05). Postoperative complication variables showed that there was no significant difference in the number of patients who suffered from cardiac events and stroke (2.4% CCAB vs 1.4% OPCAB, p = NS) whereas there was a significant difference for minor neurological events (12.2% CCAB vs 2.8% OPCAB, p = 0.04). The mean follow up was 2.1 +/- 1.8 years, for a total follow-up of 234 patient-years. There were 12 late deaths, 6 in the CCAB group and 6 in the OPCAB group. The linearized rate of overall death was 9.5 +/- 2.6%/year in the CCAB group and 13.3 +/- 3.9%/year in the OPCAB group (p = NS). The actuarial survival at 24 months was 84 +/- 7% for OPCAB and 81 +/- 12% for CCAB (p = NS). The actuarial freedom from cardiac events at 24 months was 89 +/- 4% for OPCAB and 90 +/- 6% for CCAB (p = NS). The linearized rate of neurological events was 0.7 +/- 0.7%/year for the CCAB group and 1.1 +/- 1.1%/year for the OPCAB group (p = NS). CONCLUSIONS: Our analysis shows that OPCAB improves the early outcome and that octogenarians can lead event free lives after cardiac surgery. PMID- 15119503 TI - Atrial fibrillation requiring urgent medical care. Approach and outcome in the various departments of admission. Data from the atrial Fibrillation/flutter Italian REgistry (FIRE). AB - BACKGROUND: The atrial Fibrillation/flutter Italian REgistry (FIRE) study was designed to obtain updated information regarding the clinical characteristics of and medical approach to patients requiring urgent medical care for atrial fibrillation (AF) or atrial flutter in a nationwide and representative series of hospitals. METHODS: 4570 consecutive patients admitted to the emergency room for AF/atrial flutter were enrolled in 207 hospitals. Of these, 2838 (61.9%) were hospitalized (median 6 days, 43% in cardiology and 57% in internal medicine departments), and constitute the population of this study. RESULTS: AF/atrial flutter represented 1.5% of all emergency room admissions and 3.3% of all hospitalizations. The mean age was 70 +/- 12 years; 89.9% had AF and 10.1% atrial flutter. In 31% of the hospitalized patients no cardiac disease was present, and in 18% no disease (either cardiac or non-cardiac) could be detected. Predictors of no attempt of cardioversion (37.5% of patients) included: onset of AF > 48 hours, heart failure, increasing age, syncope, admission to a non-cardiology department, stroke or transient ischemic attack (TIA). Predictors of in-hospital mortality (2.2%) included: age, heart failure, diabetes, admission to a non cardiology department, and stroke or TIA. Predictors of the absence of sinus rhythm at discharge (35.6% of patients) included: no attempt of cardioversion, heart failure, chronic anticoagulation, AF duration > 48 hours, increasing age, stroke or TIA, and admission to a non-cardiology department. Transesophageal echocardiography was performed in only 6% of patients. CONCLUSIONS: AF/atrial flutter represent a significant burden on the health care system with a higher than expected hospitalization rate from the emergency room. One out of three discharged patients is not in sinus rhythm. There is still a wide gap between evidence-based medicine and real practice in the treatment of patients with AF. PMID- 15119504 TI - Electrocardiographic features in critical pulmonary embolism. Results from baseline and continuous electrocardiographic monitoring. AB - BACKGROUND: Pulmonary embolism (PE) afflicts millions of individuals worldwide. Electrocardiography along with chest X-ray and arterial blood gas analysis represent the basic examinations to reinforce the clinical suspicion of PE. We describe the electrocardiographic (ECG) features in a series of patients with PE and a critical clinical presentation. METHODS: We report the ECG findings registered at baseline, 48 hours after admission and on continuous ECG monitoring in 51 patients with PE and critical clinical conditions. RESULTS: At admission, the following parameters were recorded: an S1Q3 pattern in 34 patients, a "septal embolic pattern" in 27, anterior lead T-wave inversion in 8, and a new right bundle branch block in 7. At 48 hours after admission a trend toward a regression of the S1Q3 and "septal embolic" patterns was noted together with evident T-wave inversion in the anterior leads. During continuous ECG monitoring no major arrhythmias were recorded, even in case of cardiopulmonary arrest. CONCLUSIONS: Critical PE induces transient ECG abnormalities reflecting right ventricular overload and/or strain. The patient's clinical status is usually not complicated by major ventricular arrhythmias, not even in case of cardiopulmonary arrest. PMID- 15119505 TI - Operations on the thoracic aorta and antegrade selective cerebral perfusion: our experience with 462 patients. AB - BACKGROUND: We retrospectively analyzed the hospital mortality and neurologic outcome after surgery on the thoracic aorta with the aid of antegrade selective cerebral perfusion to determine a predictive risk model. METHODS: Between October 1995 and May 2002, 462 patients (mean age 62.7 +/- 11.7 years) underwent surgery on the thoracic aorta using antegrade selective cerebral perfusion. The indication for surgery was acute type A dissection in 132 patients (28.6%), degenerative aneurysm in 258 (55.8%), and post-dissection aneurysm in 72 (15.6%). One hundred and forty-one patients (30.5%) were operated on urgently; concomitant procedures were performed in 190 patients (41.1%). The mean cerebral perfusion time was 63 +/- 39 min. Predictors of hospital mortality and neurologic outcome were identified by univariate and multivariate analysis of the preoperative and intraoperative variables. RESULTS: The hospital mortality rate was 10.2%. Stepwise logistic regression identified an urgency status (odds ratio--OR 5.2, p = 0.001), a history of a central neurologic event (OR 4.1, p = 0.007) and coronary artery bypass graft (OR 3.2, p = 0.039) as being independent determinants for hospital mortality. The transient neurologic dysfunction rate was 6.2%. An urgency status (OR 3.4, p = 0.003) and a history of a central neurologic event (OR 5.1, p = 0.002) were independent determinants of transient neurologic dysfunction. An urgency status (OR 6.0, p = 0.011) was the only independent determinant for permanent neurologic dysfunction (3.8%). A cerebral perfusion time > 90 min was not associated with an increased risk of hospital mortality and permanent or transient neurologic dysfunction. CONCLUSIONS: Antegrade selective cerebral perfusion proved to be a safe method of brain protection allowing complex aortic repair to be performed with encouraging results in terms of hospital mortality and neurologic outcome. PMID- 15119506 TI - Global cardiovascular risk evaluation in Italy: a cross-sectional survey in general practice. AB - BACKGROUND: The aim of our work was to evaluate, in a general practice setting, the attitude of general practitioners in determining the individual coronary risk. METHODS: The coronary risk was determined among patients aged 30 to 74 years using the following parameters: gender, age, smoking habits, diagnosis of diabetes mellitus, systolic blood pressure, and total cholesterol. We evaluated the records of 446,331 subjects collected by 481 general practitioners working throughout Italy. RESULTS: Except for age, gender and diabetes mellitus, risk factors were largely under-recorded: blood pressure in 37.0% of the total patients, total cholesterol in 34.3%, smoking habits in 21.9%. Recording was substantially low even in patients who were prescribed with antihypertensive drugs and/or lipid-lowering drugs: blood pressure in 80.6% of the patients, total cholesterol in 69.1%, smoking habits in 46.1%. Cardiovascular risk factors were more frequently recorded as age increased and slightly more among women as compared to men. Obviously, it is possible that risk factors had been assessed but not recorded. CONCLUSIONS: Cardiovascular risk factors are substantially under-recorded among Italian general practitioners thus impairing adequate preventive treatment. A systematic, well programmed approach may theoretically lead to evaluate the majority of the target population within a few years. PMID- 15119507 TI - Fulminant myocarditis during HIV seroconversion: recovery with temporary left ventricular mechanical assistance. AB - A 32-year-old male was admitted to our intensive care unit for low cardiac output syndrome. Echocardiography was suggestive of extensive hypokinesia and the ejection fraction was 0.22. Serological tests, including anti-HIV antibodies (ELISA), were negative. The patient was intubated and an intra-aortic balloon pump was inserted. Twenty-four hours after admission a paracorporeal left ventricular assist device (LVAD-MEDOS) was implanted. The left ventricular function showed progressive improvement with normalization of the ejection fraction on day 19. The device was removed on day 20. Before discharge, the patient admitted that he had had unprotected sex with numerous male acquaintances; anti-HIV testing turned positive. The final diagnosis was fulminant myocarditis during HIV seroconversion. PMID- 15119508 TI - Main pulmonary artery aneurysm: a case report and review of the literature. AB - Main pulmonary artery aneurysms are a rare entity with few available published data. As reported in the literature, operative treatment is commonly recommended but the relation between the size of the aneurysm, its localization, and the risk of rupture is not as well defined as for aortic aneurysms. Proximal lesions that involve the main branches of the pulmonary artery are usually apparent on chest radiographs and must be taken into consideration in the differential diagnosis of mediastinal masses. An early diagnosis allows timely surgical treatment. We report an unusual case of a main pulmonary artery aneurysm presenting with persistent non-productive cough and provide a review of the pertinent published data. PMID- 15119509 TI - The triple-orifice repair: a new technique for the treatment of mitral regurgitation in severe Barlow's disease. AB - We report a case of severe Barlow's disease with a very complex pathology, in which we applied the "edge-to-edge" technique, creating a triple-orifice mitral valve. Different techniques should be used to correct a similar valve defect; the combination of different surgical procedures and the valve pathology may influence the post-repair recurrence of regurgitation. We believe that it is better to perform a simple and reproducible repair than to carry on with combined complex procedures that could increase the risk of a suboptimal outcome. PMID- 15119510 TI - Upper extremity deep vein thrombosis and pulmonary embolism after coronary bypass surgery: a case report and preliminary results from a prospective study evaluating patients during cardiac rehabilitation. AB - A 78-year-old woman with unstable angina underwent coronary bypass surgery with complete cardiac revascularization and no immediate postoperative complications. Six days after surgery, during hospitalization for cardiac rehabilitation, the patient developed severe respiratory distress and pulmonary embolism was diagnosed. Color duplex ultrasound revealed the presence of concomitant upper extremity deep vein thrombosis (UEDVT), ipsilateral to the site of placement of a central venous line, in the absence of lower extremity deep vein thrombosis. We describe this case and provide preliminary data from a prospective observational study evaluating the prevalence of catheter-related UEDVT and symptomatic pulmonary embolism (55 and 1.4% respectively) in a series of 71 consecutive coronary bypass surgery patients admitted to a cardiac rehabilitation facility. Catheter-related UEDVT and pulmonary embolism may complicate coronary bypass surgery and should be taken into consideration when managing patients after surgery. PMID- 15119511 TI - Echo-guided surgical anteroseptal left ventricular restoration for end-stage idiopathic dilated cardiomyopathy. PMID- 15119512 TI - Guidelines--let's take a break and then move forward together! AB - With our limited resources, the national colleges of general practice in the Nordic countries should select only a few important topics and only get involved in guideline preparations if they have a primary care perspective. We need not develop our own guidelines from scratch, but should take existing international GP clinical guidelines and literature reviews as a starting point and conduct the first steps in the guideline preparation jointly in the Nordic countries. More effort could then be directed towards the subsequent stages of guideline preparations and the implementation process. Key stakeholders should be involved at all stages in guideline development and at all levels of the health services to ensure commitment and improve the likelihood of implementation. PMID- 15119513 TI - Collection and retrieval of structured clinical data from electronic patient records in general practice. A first-phase study to create a health care database for research and quality assessment. AB - OBJECTIVE: To evaluate prerequisites, practicalities, attitudes and limitations related to the collection of structured clinical data in everyday general practice for use in the future establishment of a national registration network. DESIGN: Prospective study. SETTING: Primary health care centres in south-western Sweden. SUBJECTS: Fourteen participating general practitioners in five primary health care centres. MAIN OUTCOME MEASURES: Feasibility and workload involved in structured data entry and in the retrieval of data from different record systems. The accuracy of clinical data in terms of clinical variables, correctness and representativeness. RESULTS: All four record systems could deliver basic data on the patient population. One centre had to be excluded from further data retrieval because of limitations in the data retrieval export format. Collecting data in everyday practice was feasible with acceptable data accuracy and moderate workload. CONCLUSION: It was feasible to collect, retrieve and store structured clinical data with respect to accuracy and extra workload. Interest in a national registration network and an increasing demand for information about primary health care in order to optimise clinical practices and support research, creates prerequisites for establishing a valid and reliable database. However, developmental work focusing on classification limitations, coding tools and routines for data retrieval is necessary. PMID- 15119514 TI - Positive self-assessed general health in patients with medical problems. A qualitative study from general practice. AB - BACKGROUND: Patient and doctor do not always agree on the status of the patient's health. By underestimating the patient's strong sides, the doctor may be contributing to disempowerment and bypassing knowledge needed for adequate medical diagnosis and management. OBJECTIVE: To understand how our patients with medical problems assess their general health as good. DESIGN AND SETTING: Qualitative observational study based on audiotaped material from general practice consultations in authors' practices (Norway and Denmark). The patients were asked to rate their current state of health on a scale ranging from zero to 100, and then to explain their score. SUBJECTS: From 41 consecutive consultations we compiled a purposeful sample of 12 patients who reported positive self assessed general health although medical problems were present. The 7 women and 5 men were aged between 43 and 96 years, and had been diagnosed with musculoskeletal disorders, heart disease, cancer, depression, headache or severe menopausal symptoms. MAIN OUTCOME MEASURES: Salutogenesis, represented by the authors' Health Resource/Risk Balance Model, and Antonovsky's Sense of Coherence (SOC) concept comprised the theoretical framework. Transcripts from audiotaped consultations were used for qualitative text condensation analysis, inspired by Giorgi's phenomenological method. Analysis was theory-driven, applying comprehensibility, manageability and meaningfulness as entries to elaborate patients' accounts of positive health. RESULTS: Patients' answers demonstrated how a feeling of logical reasoning related to symptom perception could provide comfort and sometimes lead to advantageous coping strategies. Personal and social resources were mentioned as essential means for tolerating and managing the burden of disease. Even fairly extensive endeavours could be experienced as worthwhile when sometimes providing relief, even only temporarily. CONCLUSIONS: Patients' accounts of general health can challenge the traditional medical views on assessment of health and disease. PMID- 15119515 TI - Recurrent antibiotic use in a small child and the effects on the family. AB - OBJECTIVE: To determine the typical characteristics of small children on recurrent therapy with antibiotics (RTA) and the effects RTA have on the families. DESIGN: Stratified randomised cluster sampling. SETTING: An unselected population-based questionnaire study in Finland. SUBJECTS: Parents (n = 1443) expecting their first child were followed prospectively from the mother's early pregnancy until the child was 18 months of age. MAIN OUTCOME MEASURES: The outcome measure was the number of courses of antibiotic therapy (options: 011-51 > or = 6) during the child's first 18 months of life. Six or more courses were considered RTA. Associated variables were child- and family-related factors during the child's first 18 months of life. RESULTS: Children on RTA were taken to see a physician more often than other children when they had fever or a common cold. RTA was significantly associated with the child's day care outside the home, allergy and need for a special diet. The father's severe stress was associated with the child's RTA. Breastfeeding lasting more than 3 months was found to have a protective effect against RTA. CONCLUSION: The threshold for seeking medical help was low in the families of children on RTA, which is best avoided by breastfeeding and day care at home. Health care staff should not forget to support families, especially the fathers, with children experiencing recurrent infections. PMID- 15119516 TI - Patients' and physicians' views on the management of acute maxillary sinusitis. AB - OBJECTIVES: To study the views of patients and physicians on the management of suspected acute maxillary sinusitis and on suggested changes in practice. DESIGN: Cross-sectional survey and focus group interviews of both patients and physicians. SETTING: Primary care in Finland. RESULTS: A total of 175 patients and 245 physicians replied to the survey (response rates 88% and 70%, respectively). Eight focus groups were conducted, four with patients (n = 22) and four with physicians (n = 20). Patients had less trust than physicians in the natural healing process of acute maxillary sinusitis: only 33% as opposed to 95% of physicians agreed that acute maxillary sinusitis might heal without medication. The patients emphasised the importance of careful diagnosis. Physicians experienced the diagnostic uncertainty and the demands from authorities to use fewer antibiotics as problematic. CONCLUSIONS: Most patients with suspected acute maxillary sinusitis primarily want the physician to aim at making a specific diagnosis. Physicians feel they are forced to prescribe too many antibiotics for sinusitis because of patient's expectations and lack of good diagnostic means for acute maxillary sinusitis. Better diagnostic tools, both national and local guidelines, and patient information might help to change practice towards the use of fewer antibiotics. PMID- 15119517 TI - Gender differences in risk factor control and treatment profile in diabetes: a study in 229 swedish primary health care centres. AB - OBJECTIVE: To measure the risk factors and treatment profile of diabetes patients treated in primary health care (PHC) in order to evaluate potential gender differences. DESIGN: Cross-sectional survey of consecutive diabetes patients. SETTING: 229 PHC centres in Sweden. SUBJECTS: 5082 men and 4293 women with diabetes were investigated (1998-2001). MAIN OUTCOME MEASURES: Glycaemic control (HbA1c), blood pressure, lipid levels, prevalence of left ventricular hypertrophy and microalbuminuria. Proportions of patients with previous ischaemic heart disease (IHD) and specific drug treatment. RESULTS: Male patients generally had better blood pressure ( < 140 and/or 85 mmHg) and glycaemic (HbA1c < 6.5%) control than corresponding female patients (44% and 59%, versus 40% and 54% in the 60-75 year age group; p < 0.01). Females showed higher levels of total (p < 0.01) and HDL cholesterol (p < 0.05) than males in all age groups. No gender difference was detected for LDL cholesterol levels in the younger or elderly patients, but in the age group 60-75 years female patients had significantly higher mean LDL cholesterol level than male patients (3.3 vs 3.2 mmol/L; p < 0.05). Previously known manifestations of IHD were more common (p < 0.01) in male patients. CONCLUSION: Elderly male patients with diabetes had a more favourable risk factor control than corresponding female patients. PMID- 15119518 TI - Barriers to implementing cardiovascular risk tables in routine general practice. AB - DESIGN: Qualitative study. GPs were interviewed after analysing two audiotaped cardiovascular consultations. SETTING: Primary health care. SUBJECTS: A sample of 15 GPs who audiotaped 22 consultations. MAIN OUTCOME MEASURES: Barriers hampering GPs from following the guideline. RESULTS: Data saturation was reached after about 13 interviews. The 25 identified barriers were related to the risk table, the GP or to environmental factors. Lack of knowledge and poor communication skills of the GP, along with pressure of work and demanding patients, cause GPs to deviate from the guideline. GPs regard barriers external to themselves as most important. CONCLUSION: Using the risk table as a key element of the high-risk approach in primary prevention encounters many barriers. Merely incorporating risk tables in guidelines is not sufficient for implementation of the guidelines. Time-efficient implementation strategies dealing in particular with the communication and presentation of cardiovascular risk are needed. PMID- 15119519 TI - The first general practitioner hospital in The Netherlands: towards a new form of integrated care? AB - OBJECTIVE: To describe the types of patients admitted to the first Dutch general practitioner (GP) hospital, their health-related quality of life and its substitute function. DESIGN: A prospective observational study. SETTING: The remaining 20-bed ward of a former district general hospital west of Amsterdam; a region with 62000 inhabitants and 26 GPs. SUBJECTS: All patients admitted during the 12 months between 1 June 1999 and 1 June 2000. MAIN OUTCOME MEASURES: Patients' health-related quality of life (Medical Outcome Study 36-item Short Form Health Survey, Groningen Activities Restriction Scale), GPs assessments of severity of illness (DUSOI/WONCA Severity of Illness Checklist) and alternative modes of care. RESULTS: In total, 218 admissions were recorded divided into 3 bed categories: GP beds (n = 131), rehabilitation beds (n = 62) and nursing home beds (n = 25). The mean age of all patients was 76 years. Main reasons for admission were immobilization due to trauma at home (GP beds), rehabilitation from surgery (rehabilitation beds) and stroke (nursing home beds). Overall, patients showed a poor health-related quality of life on admission. If the GP beds had not been available, the GPs estimated that the admissions would have been almost equally divided among home care, nursing home and hospital care. The severity of the diagnosis on admission of the 'hospital-care group' appeared to be significantly higher than the other care groups. CONCLUSION: The GP hospital appears to provide a valuable alternative to home care, nursing home care and hospital care, especially for elderly patients with a poor health-related quality of life who are in need of short medical and nursing care. PMID- 15119520 TI - Deep venous thrombosis: a new task for primary health care. A randomised economic study of outpatient and inpatient treatment. AB - OBJECTIVE: A health economic evaluation of two alternative treatment settings, inpatient care and outpatient care, for acute deep venous thrombosis. DESIGN: A randomised multicentre trial in a defined population in regular clinical practice. SETTING: Hospitals and related health care centres in the Jonkoping county council in Sweden. INTERVENTIONS: Patients were randomised to either an inpatient strategy (n = 66) or an outpatient strategy (n = 65) using low molecular-weight heparin, dalteparin, administered subcutaneously once daily and adjusted for body weight. SUBJECTS: Of 224 eligible patients, 131 entered the trial and 124 completed the economic part of the study. MAIN OUTCOME MEASURES: Direct medical and direct non-medical costs during a 3-month period. RESULTS: Total direct costs were higher for those in the inpatient strategy group, i.e. Swedish Crowns (SEK) 16400 per patient (Euro 1899) compared to SEK 12100 per patient (Euro 1405) in the outpatient strategy group (p < 0.001). More patients in the outpatient group received assistance when they returned home. Few patients in either group reported sick leave. There was no difference in total number of days between the two groups. CONCLUSIONS: Total direct costs were significantly lower for the outpatient treatment strategy for deep venous thrombosis compared to the inpatient treatment strategy. No significant difference in health impact could be detected. Deep venous thrombosis can to a greater extent than previously be treated in primary care, safely, at a lower cost, and in accordance with patient preferences. PMID- 15119521 TI - Adolescent tobacco smoking and associated psychosocial health risk factors. AB - OBJECTIVE: To determine the relationship between tobacco smoking and psychosocial health risk factors in adolescents. DESIGN: An in-class survey of schoolchildren in the final grade of basic school and in all three grades of secondary school. SETTING: Four of the 11 secondary schools in Tartu, Estonia. SUBJECTS: 977 schoolchildren, ages ranging from 14 to 18. MAIN OUTCOME MEASURES: A questionnaire was developed to explore various psychosocial aspects of tobacco smoking behaviour. The 21-item Beck Depression Inventory was used to measure depression. RESULTS: In the total sample, smokers accounted for 24.5% of the girls and 26.5% of the boys, of which 13% of the girls and 19% of the boys were daily smokers. Not enjoying time spent with parents (OR = 0.6), skipping breakfast (OR = 1.3), frequent headache (OR = 1.3) and stomach-ache (OR = 1.4), dislike of school (OR = 0.7), using illicit drugs (OR = 5.0) and having multiple sexual partners (OR = 2.4) were all associated with daily smoking. Higher BDI scores were seen among adolescent smokers, particularly in girls and among pupils whose parents were non-smokers. The girls who smoked daily showed a higher risk of having suicidal thoughts (OR = 2.4) compared with non-smokers. CONCLUSION: Smoking by adolescents is an indicator of risk for depression, distress and risk taking health-damaging behaviours. PMID- 15119522 TI - Consultation with the general practitioner triggered by advice from social network members. AB - OBJECTIVE: To investigate whether advice from a person's social network triggers contact with the general practitioner (GP). DESIGN: Case-crossover design comparing the frequency of advice given to seek medical attention in the period before contact with a GP and the frequency in matching control time periods for the same individual. SETTING: Twenty-one Danish GPs working in single-handed practices. SUBJECTS: 322 patients, aged between 18 and 91 years, were interviewed by telephone after an unscheduled visit to their GP; 148 were interviewed again 3 6 months later. MAIN OUTCOME AND MEASURES: The odds of individuals consulting their GP after receiving advice from network members in the period before they contacted their GP compared with the odds of those consulting their GP in the control period(s). RESULTS: Being advised by others to seek medical attention increased the likelihood of seeking primary health care approximately fivefold- single men received advice significantly less frequently (7%) than women (18%) and cohabiting men (32%). CONCLUSION: Advice from other social network members to seek medical attention is a frequent and influential cue prompting individuals to contact their GP. PMID- 15119523 TI - Estimating incidence and prevalence of episodes of care in general practice. AB - OBJECTIVES: To develop methods for prospective registration and analysis of episodes of care (one or more contacts about the same health problem). To compare estimates of point prevalence and incidence of hypertension among a group of elderly patients by means of an elaborate coding of episodes and a new method based on routine coding of contact diagnoses. DESIGN: Prospective longitudinal cohort study. SETTING: General practice in Denmark. SUBJECTS: 31 GPs and 1722 patients 70+ years of age. MAIN OUTCOME MEASURES: During the course one year, data were registered in the doctors' electronic medical record systems. Registration included ICPC diagnoses and codes for diagnostic and therapeutic procedures. Individual contacts were linked into episodes of care by the recording physician. Prevalence and incidence were calculated from the GPs' registration of episodes of care and from contact registration by means of a new method, "the waiting time distribution". RESULTS: Estimates of incidence (2.1 per 100 patient years) and point prevalence (21.2 per 100 patients) of episodes of hypertension care could be calculated when the GPs actively linked contacts into episodes. Based on simple contact registration it was possible to calculate similar estimates. CONCLUSION: A full registration of episodes of care is time consuming and complicated. Incidence and point prevalence of chronic conditions, however, may be estimated from simple coding of contact diagnoses. PMID- 15119524 TI - Comments on: Brautbar, N. and Howard, J. 2002: Phosphine toxicity: report of two cases and review of the literature. Toxicology and Industrial Health 18, 71-75. PMID- 15119525 TI - A quantitative model of cellular senescence influence on cancer and longevity. AB - Contrary to the paradigm that cancer incidence increases indefinitely with age, significant data now suggest cancer incidence may markedly reduce beyond age 80 years for humans and beyond 800 days for mice, and is not inevitable. We show that increasing cellular senescence with age is a possible cause of this reduction, since senescent cells are removed from the pool of cells that retain proliferative ability necessary for cancer. We further show that animal interventions appearing to alter senescence, p53 mutation and melatonin dosing, support the prediction that increasing senescence rate reduces cancer while reducing lifespan, and vice versa. Studies of environmental agents associated with increased cancer might be re-examined to find if there is an association with longevity increases, which may markedly alter our view of such agents. We also show that if an agent functions by slowing both senescence and carcinogenesis, longevity is increased while reducing cancer. Dietary restriction is the only known intervention that accomplishes this, but there may be others. PMID- 15119526 TI - Effect of mercury vapour exposure on urinary excretion of calcium, zinc and copper: relationship to alterations in functional and structural integrity of the kidney. AB - BACKGROUND: The kidney has a remarkable capacity to concentrate mercury (Hg) and as such is a primary target organ when exposure to Hg occurs, and it is also an organ for Hg excretion. OBJECTIVE: The present work aims to investigate the effect of occupational Hg vapour exposure on the urinary excretion of calcium (Ca), zinc (Zn) and copper (Cu), and the possible association of this excretion to work duration as well as renal alterations. METHODS: 83 non-smoker participants (36 referents, age: 35.6 +/- 9.5 years; 27 Hg vapour-exposed workers with < or = 10 years work duration, age: 33.0 +/- 5.1 years; and 20 Hg vapour exposed workers with > or = 11 years work duration, age: 39.50 +/- 8.50 years) were included in the present study. Urinary levels of microalbumin (U-Malb) and retinol-binding protein (U-RBP) as well as cytosolic glutathione S-transferase activity (U-GST) were measured to assess the glomerular and proximal tubular reabsorption functions as well as structural integrity of proximal tubules; respectively. In addition, blood Hg (B-Hg), serum levels of Hg (S-Hg) and Ca (S Ca), and urinary levels of Hg (U-Hg), Ca (U-Ca), Zn (U-Zn), Cu (U-Cu) and creatinine (U-cr) were estimated. RESULTS: In comparison to referents, all investigated parameters showed significant increase (except S-Ca and U-Zn/U-Cu ratio that significantly decreased among the workers as one group, S-Ca and U Zn/U-Cu ratio that significantly and nonsignificantly decreased; respectively among workers with < or = 10 years work duration, S-Ca and U-Zn/U-Cu ratio that significantly decreased among workers with > or = 11 years work duration). In addition, B-Hg was nonsignificantly increased and S-Ca was significantly decreased; also, both U-Hg and U-Zn/U-Cu were nonsignificantly decreased among workers with > or = 11 years work duration in comparison to those with < or = 10 years work duration. Also, each of U-Hg, U-Ca, U-Zn and U-Cu was related to one another, while each of U-Ca, U-Zn and U-Cu was related to each of U-Malb, U-RBP and U-GST (except U-Zn was not related to U-GST). CONCLUSION: Hg vapour exposure leads to renal alterations which may parallel the change in proteinuria and enzymuria as well as the increased loss in urine of each of Ca, Zn and Cu. The urinary assessment of these metals may be used as a good indicator for renal dysfunction. PMID- 15119527 TI - Therapeutic effects of glucose infusion on monochloroacetic acid exposure in rats. AB - The effects of glucose infusion on monochloroacetate (MCA) exposure were examined in male rats with a view toward effective clinical treatment for MCA intoxication. Rats were injected with 80 mg/kg sodium monochloroacetate (SMCA) (single lethal dose) and then infused with saline (control group) or 5% or 10% glucose solution at 2 mL/hour for ten hours. No animal in the control group survived the total 14-day follow-up period. The survival rate in 5% glucose group was 57% at ten hours; it decreased to 14% at 14 days. The survival rate in 10% glucose group was 79% at ten hours, and all rats that survived the first ten hours also survived the 14 days. Kaplan-Meier analysis showed the survival rate in 10% glucose group to be improved upon in both the 5% glucose group and the control group. Blood glucose and lactate levels were measured every hour during infusion. Blood glucose levels decreased in the control group but remained in the glucose-infused groups. Although the blood lactate level increased in each group, there was an excellent inverse linear relation between blood glucose levels and blood lactate levels. Thus, continuous parenteral infusion of glucose solution at an early stage after exposure may be an effective clinical therapy for the prevention of hypoglycaemia and metabolic lactic acidosis caused by MCA. PMID- 15119528 TI - In vivo ingestion of heavy metal particles of Se, Hg and W by murine macrophages. A study using scanning electron microscopy coupled with X-ray microanalysis. AB - Several heavy metals that are currently employed in industry may become polluters of work and natural environments. As particulate matter, heavy metals are suitable for entering the human body through the respiratory and digestive systems. They often end up inside phagocytes; the size of the microscopic particles modulates both their phagocytosis, and the physiology of macrophages. Here we have adopted an experimental model to investigate the ingestion of particles of three industrial heavy metals (Se, Hg, W) by murine peritoneal macrophages in vivo. The phagocytes were studied by scanning electron microscopy coupled with X-ray elemental microanalysis (SEM-XRM), a method that allows specific identification of Se, W and Hg in cells at high resolution. We found that Hg that was taken up by macrophages was organized into small, round particles (0.31 +/- 0.14 microm). This was in contrast with the larger size of intracellular particles of Se (2.37 +/- 1.84 microm) or W (1.75 +/- 1.34 microm). Ingested particles of Se and W, but not Hg, often caused bulging of the cell surface of macrophages. We conclude that particulate matters of Se, W and Hg are organized in particles of different size inside macrophages. This size difference is likely to be associated with distinct phlogistic activities of these heavy metals, Se and W causing a milder inflammatory reaction than Hg. PMID- 15119529 TI - Dry deposition (downward, upward) concentration study of particulates and water soluble ionic species during daytime, night-time period at the traffic sampling site of Sha-Lu, Taiwan. AB - Ambient suspended particulate (dry deposition, TSP) was collected in the traffic sites Sha-Lu, central Taiwan. In addition, the related water-soluble ionic species (Cl-, NO3(-), SO4(2-), Na+, NH4(+), K+, Mg2+ and Ca2+) were analysed and wind speed, wind direction and temperature were also measured in this study. The downward dry deposition fluxes (averaged 54.07 microg/m2-sec) were about twice that of upward dry deposition fluxes (averaged 26.48 microg/m2-sec) in the daytime period. Furthermore, the average downward dry deposition fluxes (averaged 26.22 microg/m2-sec) were also about twice that of upward dry deposition fluxes (averaged 12.11 microg/m2-sec) in the night-time period. The results showed that the total suspended particulate concentrations of particulate mass in the daytime period (averaged 996.2 microg/m3) were higher than in the night-time period (averaged 560.7 microg/ m3). The results showed that the total suspended particulate concentrations of particulate mass in the daytime period (averaged 996.2 microg/m3) were higher than in night-time period (averaged 560.7 microg/ m3). As for water-soluble ionic species, the average dry deposition order and velocity for downward ionic species were Cl- > Ca2+ > NO3(-) > K+ (2.09 cm/sec > 1.46 cm/sec > 1.46 cm/sec > 1.07 cm/sec) anions during the daytime period. And the average dry deposition order and velocity for downward ionic species were NO3(-) > Cl- > K+ > Ca2+ (2.92 cm/sec > 2.74 cm/sec > 0.96 cm/sec > 0.93 cm/sec) anions during the night-time period. The average dry deposition order and velocity for upward ionic species were Cl- > Ca2+ > K+ > Mg2+ (4.69 cm/sec > 0.62 cm/sec > 0.59 cm/sec > 0.55 cm/sec) anions during the daytime period. And the average dry deposition order and velocity for upward ionic species were Cl- > Ca2+ > Mg2+ > K+ (1.65 cm/sec > 0.43 cm/sec > 0.37 cm/sec > 0.33 cm/sec) anions during the night-time period. The results also indicated that the sodium and chloride concentrations in total suspended particulate were highly positively related, indicating that the sea-salt aerosols were the major contributors for these species at this sampling site of central Taiwan. PMID- 15119530 TI - Omeprazole treatment: genotoxicity biomarkers, and potential to induce CYP1A2 activity in humans. AB - Omeprazole is one of the most used acid-suppressing medications. This fact emphasizes the questions concerning the safety of this compound. Healthy volunteers (n=33) were included in this prospective study. All study subjects were analysed for their CYP2C19 genotype. Of the 33 individuals, 24 were homozygous for the wild type CYP2C19*1 allele, 7 were heterozygous for the CYP2C19*2 variant allele, and 2 were homozygous for the CYP2C19*2 variant allele. Before and after 14 days of omeprazole treatment at a daily dose of 20 mg, one blood sample was taken from each individual to determine five cytogenetic biomarkers of genotoxicity: chromosome aberrations, micronuclei, proliferating rate index, sister chromatid exchanges, and mitotic index. The only significant change was that of a weak increase in micronuclei count after treatment in relation to baseline values (day 0) (P = 0.026). To assess the potential of omeprazole to induce P450 CYP1A2, the urinary ratio AFMU+1X+1U/17U in the interval of 4-5 hours after caffeine intake was calculated twice (days 0 and 15), using the caffeine test in 27 of the 33 individuals. This result suggests that omeprazole does not increase CYP1A2 activity after 14 days of treatment. PMID- 15119531 TI - Risk assessment of lung cancer related to environmental PAH pollution sources. AB - We assessed the lung cancer risk in six localities with aluminium smelting activities and five with other polycyclic aromatic hydrocarbon (PAH) pollution sources, using two quantitative risk assessment (QRA) approaches for PAH mixtures and compared their risk predictions against actual cancer incidence. In the first approach, carcinogen exposure was estimated from animal-derived BaP toxic equivalents (BaPeq) of individual PAHs. The upper bound lifetime risk estimates ranged between 0.012-4.7 x 10(-5) and 0.019-0.94 x 10(-5) in the aluminium and other localities, respectively. The second approach assumed that the potency of PAH mixtures was linked to their BaP content and lifetime lung cancer unit risk gradients were estimated from epidemiological studies based on BaP exposure measurements. Lifetime risks ranged between 0.02-89 x 10(-5) and 0.06-6.8 x 10( 5) in the aluminium and other localities, respectively. Predicted risks were generally higher in smelter towns, and higher when based on epidemiological studies than on BaPeq. In smelting communities, there was a linear relationship (R2 approximately 0.8) between female lung cancer rates and PAH exposure estimates. To conclude, animal/BaPeq-based QRAs predicted lower risks than occupational/BaP-based QRAs. Epidemiological validation of the QRA could be performed for elevated past exposure to PAHs, but not for currently lower concentrations. PMID- 15119532 TI - Effect of lipoic acid on the oxidoreductive status of red blood cells in rats subject to oxidative stress by chronic administration of adriamycin. AB - One of the most intriguing phenomena observed during adriamycin (ADR) toxicity has been attributed to ADR-induced oxidative stress. The study was aimed to assess the protective effect of lipoic acid (LA) against ADR-induced damage to erythrocytes. Male albino rats (Wistar strain) were subjected to ADR (1 mg/kg body weight/day i.v.) once a week for a period of 12 weeks. Haematological indices like haemoglobin levels (Hb) and haematocrit (Ht) were also lowered along with a marked increase in the activities of serum glutamate pyruvate transaminase (SGPT) and serum glutamate oxaloacetate transaminase (SGOT). These rats demonstrated enhanced erythrocyte membrane lipid peroxidation (LPO) and an onslaught in the antioxidant defence armoury, witnessed by lowered activities of superoxide dismutase (SOD), glutathione peroxidase (GPx), vitamin A, vitamin C and vitamin E. Rats administered with ADR showed a marked decline in the activities of membrane-bound ATPases. Abnormal LPO and decreased deformability led to increased osmotic fragility of the red blood cells. Pretreatment with LA (35 mg/kg body weight/day i.p.) 24 hours prior to the administration of ADR once a week for a period of 12 weeks was effective in counteracting these biochemical disturbances, thereby minimizing the toxic side effects of ADR. PMID- 15119533 TI - Nephrotoxicity and its prevention by catechin in ferric nitrilotriacetate promoted oxidative stress in rats. AB - Intraperitoneal injection of ferric nitrilotriacetate (Fe-NTA) to rats and mice results in iron-induced free radical injury and cancer in kidneys. This study was designed to investigate the effect of catechin, a bioflavonoid with antioxidant potential, on Fe-NTA-induced nephrotoxicity in rats. Four groups were employed in the present study. Group I served as control group, Group II animals received Fe NTA (8 mg iron/kg body weight i.p.), Group III animals were given 40 mg/kg catechin p.o. twice a day for 4 days and on the 5th day Fe-NTA was challenged, and Group IV animals received catechin alone for 4 days. Renal function was assessed by measuring plasma creatinine and blood urea nitrogen. The oxidative stress was measured by renal malondialdehyde levels, reduced glutathione levels and by enzymatic activity of catalase, glutathione reductase and superoxide dismutase. One hour after a single intraperitoneal (i.p.) injection of Fe-NTA (8 mg iron/kg), a marked deterioration of renal architecture, renal function and severe oxidative stress was observed. Pretreatment of animals with catechin markedly attenuated renal dysfunction, reduced elevated thiobarbituric acid reacting substances (TBARS), restored the depleted renal antioxidant enzymes and normalized the renal morphological alterations. These results clearly demonstrate the role of oxidative stress and its relation to renal dysfunction, and suggest a protective effect of catechin on Fe-NTA-induced nephrotoxicity in rats. PMID- 15119534 TI - Delayed toxicity following acute ingestion of valpromide. AB - As valpromide is a prodrug of valproic acid (valproate), the clinical presentation of overdoses with either valpromide or valproate sodium is generally considered similar. Whereas plasma peak levels and signs of central nervous system depression occur within a few hours after the acute ingestion of regular release forms of valproate sodium, delayed toxicity and time to peak levels following valpromide ingestion can be seen as shown by the three reported cases. They were initially considered as mild because patients presented with no or only moderate symptoms and serum valproate levels were below or at therapeutic levels on admission more than 3 hours post-ingestion in two of the three patients. Serum valproate levels were not monitored until marked deterioration more than 10 hours after ingestion. At the time of deterioration, serum valproate was at toxic level in the three reported cases. Therefore, large intake of valpromide should be closely monitored because no or moderate symptoms together with low plasma levels in the first few hours after ingestion do not exclude a subsequent severe intoxication. Despite the usual favourable outcome and the poor correlation between plasma levels and toxic symptoms, patients should not be discharged until plasma levels are documented to remain at low levels for at least 10 hours after the ingestion of valpromide and the patient asymptomatic. PMID- 15119535 TI - Effect of cannabinoid ingestion (in the form of bhang) on the immune system of high school and university students. AB - The discovery of cannabinoid receptors in the immune system and a family of endogenous ligands of these receptors provides a basis for understanding the cellular and molecular mechanisms of cannabis-induced immunotoxicity. The present study was conducted on 90 nonsmoker males of high school and university students living in Tanta city of matched age and socioeconomic lifestyle. They were divided into a control group (30 males) and a bhang user group (60 males), which used bhang by eating its sweet juice after boiling with a little water and drying in an oven, 'fola'. The bhang group was divided equally into two subgroups: subgroup 1 used bhang for 6-24 months (average 19 +/- 1.2) and subgroup 2 used bhang for 24-36 months (average 31 +/- 1.7). The immunotoxic effects of using bhang appeared in the form of a significant decrease in serum immunoglobulins (IgG and IgM), and C3 and C4 complement protein concentrations (P < 0.05). In addition, our results demonstrated a significant decrease in the absolute number of functionally different subsets of peripheral blood mononuclear lymphocytes, T and B lymphocytes and natural killer (NK) cells in bhang users as compared to controls (P < 0.05). Moreover, the fatty acid amide hydrolase (FAAH) showed significant decrease in bhang users as compared to controls and in subgroup 2 as compared to subgroup 1 (P < 0.05), indicating that the decrease in FAAH protein level is closely related to the duration of bhang use. Positive correlations were found between FAAH level and the absolute number of mononuclear cells (T, B lymphocytes and NK cells) among bhang user subgroups. The present study is the first study to report on the effect of bhang on complement proteins and immunoglobulins in humans. Our study revealed that bhang-induced immunotoxicity could be attributed to decrease in FAAH protein. PMID- 15119536 TI - Innovations in access to TB and HIV/AIDS care in sub-Saharan Africa: dynamic engagement of the private sector. PMID- 15119537 TI - Alcohol in New Zealand road trauma. AB - Alcohol-impaired driving is one of the major contributing factors to fatal and serious crashes in New Zealand. To curb the high level of road trauma resulting from drink-driving, a compulsory breath test (CBT) programme was introduced in 1993 and a supplementary road safety package (SRSP) in 1995/1996. The SRSP aimed to enhance road safety enforcement and advertising activities, and focused primarily on drink-driving and speeding. These interventions have resulted in a substantial reduction in alcohol-related road trauma. Subsequently, in 1999, the drinking age was lowered from 20 to 18 years. This paper examines the impacts of these drink-driving interventions. The analysis shows that the CBT programme and the SRSP have contributed to the reduction in alcohol-related crashes in recent years. There is also some evidence that, following the lowering of the drinking age, there has been an increase in drink-driving and subsequent alcohol-related crash involvement for drivers under 18 years. PMID- 15119538 TI - Perception-based road hazard identification with Internet support. AB - One of the most important tasks faced by highway agencies is identifying road hazards. Agencies use crash statistics to detect road intersections and segments where the frequency of crashes is excessive. With the crash-based method, a dangerous intersection or segment can be pointed out only after a sufficient number of crashes occur. A more proactive method is needed, and motorist complaints may be able to assist agencies in detecting road hazards before crashes occur. This paper investigates the quality of safety information reported by motorists and the effectiveness of hazard identification based on motorist reports, which were collected with an experimental Internet website. It demonstrates that the intersections pointed out by motorists tended to have more crashes than other intersections. The safety information collected through the website was comparable to 2-3 months of crash data. It was concluded that although the Internet-based method could not substitute for the traditional crash based methods, its joint use with crash statistics might be useful in detecting new hazards where crash data had been collected for a short time. PMID- 15119539 TI - Analysis of design attributes and crashes on the Oregon highway system. AB - This paper estimates the relationship between crash activity and roadway design attributes on the Oregon state highway system. Crash models are estimated from roadway segments distinguished by functional classification and location. A number of design attributes were found to be related to crash activity in the various models, including number of lanes, curve characteristics, vertical grade, surface type, median type, turning lanes, shoulder width and lane width. In selected instances, crash reduction factors (CRFs) calculated from model results are compared with those presently used to evaluate projects in the Oregon Department of Transportation's Safety Improvement Program. The number of design attributes specified in the crash models is limited in comparison with the number of CRFs presently used to evaluate safety improvement projects. However, the attributes included in the crash models represent countermeasures associated with the more costly outlays that states make to improve safety. Thus, crash models of this type provide states with an opportunity to validate the CRFs that are most important economically. PMID- 15119540 TI - Stated preference methods in health care evaluation: an emerging methodological paradigm in health economics. AB - One focus of health economics is the trade-off between limited resources and the (health) needs of a community. Cost-effectiveness analysis (CEA), while being one of the most accepted evaluation methodologies in health economics, does not account for many important costs and benefits of health care interventions. Some health economists have attempted to modify CEA to account for these deficiencies, while others have been working on alternative methodologies. One group of alternative methodologies can be described as stated preference techniques. These aim to measure both health and non-health outcomes (ie costs and benefits), and include qualitative analysis, conjoint analysis (often referred to as discrete choice analysis/modelling) and willingness to pay (or contingent valuation). This paper provides an overview of stated preference techniques in health economics, with particular focus on their strengths as compared with traditional evaluation methods in health care. The limitations and policy implications of these methods are also discussed. PMID- 15119541 TI - County council politicians' choice of hospital payment scheme: a discrete choice study. AB - Various hospital payment schemes exist, but none of the schemes fulfil all the objectives of a public payer simultaneously. This implies that trade-offs are involved in the choice of payment scheme. The purpose of this study is to elicit Danish politicians trade-offs in terms of the objectives involved using two elicitation techniques: simple ranking and discrete choice experiments. The implied rankings of the objectives according to the two techniques are compared and reveal discrepancies. The discrepancies may be due to the nature of the techniques and the transparency of the implied ranking to the respondent. PMID- 15119542 TI - Recent advances and future perspective in neuroimaging in neuropsychiatric systemic lupus erythematosus. AB - Involvement of the central nervous system (CNS) is one of the most important complications of systemic lupus erythematosus (SLE), occurring in 14-75% of SLE patients. Neurological and psychiatric involvement is mainly manifested as cerebrovascular disease, seizures, cognitive impairment, headaches and psychosis. However, diagnosis of brain involvement in SLE (i.e., neuropsychiatric lupus: NPSLE) as well as understanding of pathogenetic mechanisms still remains a difficult challenge. Although a wide range of neurodiagnostic tools have been used in the last decade to assess CNS involvement, no single technique has proven to be definitive or reliable. Since neurometabolic impairment, neurochemistry and perfusion abnormalities in NPSLE may precede anatomic lesions, new functional techniques such as magnetic resonance spectroscopy, diffusion and perfusion weighted imaging, and magnetization transfer imaging may be useful in order to indentify pathologic changes unrevealed by conventional imaging. So these new diagnostic tools could modify diagnostic and therapeutic approaches to this major unsolved problem, also shedding some light on the physiopathology of CNS disease in SLE. PMID- 15119543 TI - Clinical significance of antinuclear antibodies in malignant diseases: association with rheumatic and connective tissue paraneoplastic syndromes. AB - Our objective was to determine the prevalence of antinuclear antibodies (ANAs) in patients with malignancies and to investigate if their presence might be related with development of musculoskeletal symptoms or paraneoplastic rheumatic syndromes. Antinuclear antibodies were determined by indirect immunofluorescence on Hep-2 cells in 274 neoplastic patients and in a control group of 140 age adjusted healthy subjects. Antinuclear antibody specificities (anti-DNA and anti ENA) were investigated in patients with rheumatological symptoms and positive ANA. Antinuclear antibodies were detected in 76 of 274 (27.7%) patients with malignancies and in nine of 140 (6.4%) healthy subjects. Twenty patients reported paraneoplastic rheumatic symptoms or syndromes. Two of them developed clinical symptoms mimicking rheumatoid arthritis (rheumatoid-like arthropathy), one systemic lupus erythematosus (lupus-like syndrome), one dermatomyositis and four cutaneous vasculitides. Musculoskeletal symptoms and paraneoplastic rheumatic symptoms and syndromes were both more frequently observed in patients with positive ANA. Antinuclear antibody specificities were found in only two cases. We can conclude that there is an increased incidence of antinuclear antibodies in malignant conditions. Musculoskeletal symptoms and rheumatic paraneoplastic symptoms and syndromes seem to be more frequent in patients with cancer-related positive ANAs. The failure to find ANA specificities (anti-ENA, anti-DNA) in patients with malignancies and positive ANAs in our study may simply reflect molecular differences between the autoantigens involved in cancer and those characteristically involved in the systemic autoimmune diseases. PMID- 15119544 TI - Absence of association between the MCP-1 gene polymorphism and histological phenotype of lupus nephritis. AB - Lupus nephritis presents two polar histological patterns, diffuse proliferative glomerulonephritis (DPGN) and membranous glomerulonephritis (MGN). In the kidney tissue of DPGN, numerous mononuclear cells were seen in the interstitium and glomeruli; on the other hand in MGN, infiltrating cells were less frequent. Monocyte chemoattractant protein-1 (MCP-1) is a potent chemoattractant for monocytes, T-cells, and natural killer cells. In this study we assessed the significance of the MCP-1 gene in determination of the histological phenotype in lupus nephritis. There was no association between the risk of DPGN and the MCP-1 gene genotype. PMID- 15119546 TI - Antinucleosome antibodies may help predict development of systemic lupus erythematosus in patients with primary antiphospholipid syndrome. AB - Patients with primary antiphospholipid syndrome (PAPS) may evolve to systemic lupus erythematosus (SLE), even many years later. This makes differentiation between primary and secondary antiphospholipid syndrome a difficult task. Studies in murine models of lupus have shown that the development of antinucleosome (anti NCS) antibodies may occur from the early stages of life. We therefore hypothesize that anti-NCS antibodies could help predict development of SLE in patients with PAPS. We studied anti-NCS antibodies in 18 PAPS patients (15 female, three male), followed for a mean of 11 years to evaluate the potential development of SLE. When PAPS was diagnosed, nine patients were positive for anti-NCS antibodies. Six of them developed clinical manifestations of SLE. In contrast, none of the patients who were negative to anti-NCS antibodies developed it. These findings suggest that anti-NCS antibodies could help predict which patients with PAPS may eventually develop SLE. PMID- 15119545 TI - Autologous stem cell transplantation for systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is responsive to treatment with immunosuppressives and steroids, but often pursues a relapsing or refractory course resulting in increasing incapacity and reduced survival. Autologous stem cell transplantation (ASCT) following immunoablative chemotherapy is a newer therapy for autoimmune disease of potential use in severe SLE. A retrospective registry survey was carried out by the European Blood and Marrow Transplant and European League Against Rheumatism (EBMT/EULAR) registry. Data was collected from 53 patients with SLE treated by ASCT in 23 centres. Disease duration before ASCT was 59 (2-155) months (median, range), 44 (83%) were female, and median age was 29 (9-52) years. At the time of ASCT a median of seven American College of Rheumatology (ACR) diagnostic criteria for SLE were present (range 2-10) and 33 (62%) had nephritis. Peripheral blood stem cells were mobilized with cyclophosphamide and granulocyte colony stimulating factor in 93% of cases. Ex vivo CD34 stem cell selection was performed in 42% of patients. Conditioning regimens employed cyclophosphamide in 84%, anti-thymocyte globulin in 76% and lymphoid irradiation in 22%. The mean duration of follow-up after ASCT was 26 (0 78) months. Remission of disease activity (SLEDAI < 3) was seen in 33/50 (66%; 95%CI 52-80) evaluable patients by six months, of which 10/31 (32%; 95%CI 15-50) subsequently relapsed after six (3-40) months. Relapse was associated with negative anti-double stranded DNA (anti-dsDNA) antibodies before ASCT (P = 0.007). There were 12 deaths after 1.5 (0-48) months, of which seven (12%; 95%CI 3-21) were related to the procedure. Mortality was associated with a longer disease course before ASCT (P = 0.036). In conclusion, this registry study demonstrates the efficacy of ASCT for remission induction of refractory SLE, although mortality appeared high. The safety of this procedure is likely to be improved by patient selection and choice of conditioning regimen. The return of disease activity in one-third of patients might be reduced by long-term immunosuppressive therapy post-ASCT. PMID- 15119547 TI - Heterogeneity of antibodies to beta2-glycoprotein 1 from patients with systemic lupus erythematosus. AB - We studied antibodies to beta2-glycoprotein 1 (anti-beta2GP1) from 72 patients with systemic lupus erythematosus (SLE) with or without antiphospholipid syndrome (APS) or with or without anticardiolipin antibodies (aCL). Fifteen patients had APS and positive antiphospholipid antibodies [clinical APS(+)/aPL(+)], 12 patients had APS, negative serum IgG and IgM aCL, antiphosphatidylethanolamine, anti-phosphatidylserine and no lupus anticoagulant [clinical APS(+)/ aPL(-)]. A third group included 16 patients without APS but high aCL levels [clinical APS( )/ aPL(+)]. In a fourth group we studied 29 patients without clinical manifestations of APS or aCL [clinical APS(-)/aPL(-)]. One hundred anticardiolipin and VDRL-negative normal sera were studied as controls. IgG antibodies to cardiolipin proper in a bovine beta2GP-free system, to human beta2GP1 immobilized on cardiolipin or to human beta2GP1 alone were detected in all sera by ELISA using irradiated and nonirradiated plates from two manufacturers. Sera from APS(+)/aPL(+) patients showed IgG binding to CL, CL + beta2GP1 and beta2GP1 in irradiated and nonirradiated plates. APS(+)/ aPL(-) sera had more significant IgG binding to beta2GP1 than normal controls when studied in both irradiated or nonirradiated plates (P = 0.001). This binding was inhibited by solid-phase cardiolipin in a dose-dependent manner. Sera from the APS( )/aPL(+) subgroup had comparable IgG activity in both the CL and CL + beta2GP1 assays, while no anti-beta2GP1 activity was detected in these sera. Sera from the clinical APS(-)/aPL(-) patients were negative in the three ELISA systems. Antibodies to human beta2GP1 from SLE patients recognize various epitopes. Those from APS(+)/ aPL(+) patients appear to react with an epitope boosted by cardiolipin in addition to another one present in the native protein. In contrast, anti-beta2GP1 from patients with APS(+)/aPL(-) are blocked by cardiolipin, suggesting that their epitope is the phospholipid-binding site. PMID- 15119548 TI - Intron 4 polymorphism of the endothelial nitric oxide synthase gene is associated with the development of lupus nephritis. AB - The objective was to investigate whether the functional polymorphism of intron 4 in the endothelial nitric oxide synthase (eNOS) gene is associated with susceptibility to systemic lupus erythematosus (SLE) and its clinical features. The 27-bp repeat polymorphism in intron 4 of the eNOS gene was determined by polymerase chain reaction in 88 SLE patients and 95 healthy control subjects. Clinical manifestations were analysed in each patient and correlated with the genotypes. The genotype distribution of the intron 4 of the eNOS did not differ between SLE patients and control subjects (aa, ab, bb genotypes 0, 15, 73 versus 2, 19, 74 controls respectively, chi-squared = 2.21, 2 df, P = 0.331). In the lupus patients according to the intron 4 genotypes of the eNOS, there was no clinically significant difference in age at onset, anti-dsDNA titre, C3, C4 level, SLEDAI, SLICC/ACR Damage Index, or autoantibodies such as RF, anti-Ro, La, RNP, Sm, or phospholipid antibodies. However, renal involvement was higher in patients with ab genotypes than in those with bb genotypes (53% versus 26%), but it did not reach statistical significance (P = 0.062). Logistic regression showed that having the ab genotype was a significant risk factor for the development of lupus nephritis (odds ratio = 3.28, 95%CI: 1.04-10.2, P = 0.04). In conclusion, our data show that the eNOS ab genotypes may be associated with the development of lupus nephritis, suggesting individuals who carry the 'a' allele are more susceptible to lupus nephritis than those with the 'b' allele. PMID- 15119549 TI - A deficiency of CD4+CD25+ T cells permits the development of spontaneous lupus like disease in mice, and can be reversed by induction of mucosal tolerance to histone peptide autoantigen. AB - It has been repeatedly shown that a subset of CD4+ T cells that constitutively express CD25 on their surface plays a role in the maintenance of self-tolerance. They may directly or indirectly affect the development of autoimmunity in susceptible mice and humans. In this study, we examine the relationship between the percentage of peripheral CD4+CD25+ T cells and the state of disease in spontaneous models of autoimmune disease. We found that both BWF1 and SNF1 mice that spontaneously develop a lupus-like disease have inherently lower percentage of the CD4+CD25+ T cells in their CD4 repertoire compared with normal Balb/c and DBA/1 mice. The percentage of CD4+CD25+ T cells was found to be increased in both normal and lupus-prone mice as they reached 7 to 8 months of age. However, mice with an autoimmune background differed from mice on a normal background in that the number of CD4+CD25+ T cells never reached 5% of the CD4 population. The lower number of the CD4+CD25+ T cells in autoimmune mice was restored to the level seen in normal mice following administration of histone peptide H471 or OVA(323-339) peptide in the absence of adjuvant intranasally but not intradermally. As such transmucosal treatment may ameliorate disease, we conclude that a deficiency in the CD4+CD25+ T cell pool contributes to a susceptibility to develop spontaneous lupus disease. PMID- 15119550 TI - Visceral leishmaniasis complicating a connective tissue disease: three case reports from Italy. AB - We report the occurrence of visceral leishmaniasis in three patients coming from the northern part of Italy treated with immunosuppressive drugs for different rheumatic conditions. This paper highlights the importance of adequate work-up in those patients presenting with serious clinical manifestations that can complicate, exacerbate or mimic a systemic connective tissue disorder. A prompt diagnosis is very important because visceral leishmaniasis has a high mortality and, if untreated, can be fatal. PMID- 15119551 TI - Systemic lupus erythematosus in Tunisia: demographic and clinical analysis of 100 patients. AB - There is a wide variation in the natural history of systemic lupus erythematosus (SLE) among different ethnic and geographical groups. Studies in Arabs are few and those in North Africans and especially in the Tunisian population do not exist. This study aims to demonstrate the demographic, clinical and laboratory characteristics of SLE Tunisian patients and to identify those at high risk for renal and neuropsychiatric involvements. One hundred patients with SLE (American College of Rheumatology criteria), seen at the Department of Internal Medicine of the University Hospital La Rabta in Tunisia over a 15-year period (1987 to 2001) were retrospectively enrolled. There were 92 women and eight men with an average age at the onset of disease of 32 years. Nineteen patients were aged over 50 years at the moment of SLE diagnosis (late-onset SLE). Of the patients, 78% had articular involvement, 53% photosensitivity and 63% malar rash. Serositis occurred in 45 patients of whom 16 had pericarditis and 29 had pleuritis. Nephritis was diagnosed in 43% of the cases and consisted always of glomerular nephritis, in three cases of which tubulointerstitial lesions were also observed. Comparison of patients with and without renal involvement showed that lupus nephritis was significantly associated with pericarditis (P = 0.03), arterial blood hypertension (P < 0.0001), cryoglobulinemia (P = 0.07) and antiphospholipid syndrome (P = 0.03). The SLEDAI at SLE diagnosis was significantly higher for lupus nephritis patients. Twelve patients with lupus nephritis died compared with three patients in the remaining group (P < 0.0001). Neuropsychiatric manifestations were observed in 25% of the cases. The mean age at SLE onset was significantly lower, the mean SLEDAI at SLE diagnosis and the mortality were significantly higher in the neuropsychiatric group than in the remaining group. Immunological features included antinuclear antibodies (100%), anti-DNA antibodies (56%), anti-Sm antibodies (61%), anticardiolipin antibodies (62%), anti-beta2GP1 (13%) anti-Rnp (23%) and hypocomplementemia (48%). The frequencies of pulmonary hypertension (25 versus 2%, P < 0.00001) and vascular thrombosis (25 versus 2%, P < 0.00001) were significantly higher in patients with positive anti beta2GP1 antibodies. The five-year survival rate in our series was 86%. The most frequent causes of death were active SLE and infections. PMID- 15119552 TI - Low-dose pulse methylprednisolone is an effective therapy for severe SLE flares. PMID- 15119553 TI - Systemic lupus erythematosus and coeliac disease. PMID- 15119554 TI - Lupus revealing itself after a stiff-person syndrome. PMID- 15119555 TI - Africa: a dentist's charitable experience of a lifetime. PMID- 15119556 TI - Initial licensure examinations. Morals and ethics. PMID- 15119558 TI - CDA defines evidence-based dentistry. Interview. PMID- 15119559 TI - Scorpion venom may hold key to stemming bone loss. PMID- 15119560 TI - The first picture of a dental forceps in a printed book. AB - Niccolo Leonico Tomeo authored numerous volumes on a variety of themes, and was principally responsible for reintroducing the works of Aristotle in the original Greek. In one of Tomeo's works, he included a picture of forceps holding an extracted tooth. This was the first time a forceps was pictured in a printed book. PMID- 15119561 TI - In the name of Dr. John Ross Callahan. AB - A look back at the original National Dental Association, illustrious researcher Dr. John Ross Callahan and some of the distinguished award recipients who exemplified his dedicated spirit. PMID- 15119563 TI - Why some of the best graduates fail the boards and why incompetent graduates are licensed. PMID- 15119562 TI - The case against one-shot testing for initial dental licensure. AB - High-stakes testing are expected to meet standards for cost-effectiveness, fairness, transparency, high reliability, and high validity. It is questionable whether initial licensure examinations in dentistry meet such standards. Decades of piecemeal adjustments in the system have resulted in limited improvement. The essential flaw in the system is reliance on a one-shot sample of a small segment of the skills, understanding, and supporting values needed for today's professional practice of dentistry. The "snapshot" approach to testing produces inherently substandard levels of reliability and validity. A three-step alternative is proposed: boards should (1) define the competencies required of beginning practitioners, (2) establish the psychometric standards needed to make defensible judgments about candidates, and (3) base licensure decisions only on portfolios of evidence that test for defined competencies at established levels of quality. PMID- 15119564 TI - Maintaining clinical excellence using SOPs. AB - Clinical excellence requires the consideration of many factors--the dentist's own level of expertise and the collaborative effort of the clinical team. The Standard Operating Procedures (SOPs) method of standardization can help a dentist move closer to that objective by allowing the completion of each task according to tested, successful protocols and can mean the difference between occasionally hitting the mark and consistently achieving greatness. PMID- 15119565 TI - The natural tooth pontic; simplified. AB - This paper's objective is to describe a simple, economical and fast method to replace a single tooth. Utilizing an indirectly fabricated composite resin framework reinforced with polyethylene fiber and the existing tooth as pontic. This tooth can be the natural tooth or the restorative crown. In addition, this particular design allows for exact repositioning of the coronal part of the extracted tooth in its original intra-oral 3-dimensional position. This chairside technique does not require laboratory involvement. PMID- 15119566 TI - Scientists espresso their glee over caffeine study. PMID- 15119567 TI - FDA to require bar coding of most pharmaceuticals by mid-2006. PMID- 15119568 TI - FDA declares actions to combat counterfeit drugs. PMID- 15119569 TI - Retrovirus conferees discuss new strategies. PMID- 15119570 TI - ASHP Health-System Pharmacy 2015 Initiative. PMID- 15119571 TI - Privileging, quality improvement, and accountability. PMID- 15119572 TI - Credentialing and privileging for pharmacists. PMID- 15119573 TI - Nutritional support and the surgical patient. AB - PURPOSE: Data on the impact of nutritional support (NS) on outcomes in surgical patients are reviewed. SUMMARY: While most patients will progress to oral nutrition after surgery and require little or no intervention, major surgery or postoperative complications can delay the prescription of an oral diet. In such patients, nutritional requirements are often increased to support wound healing and hypermetabolism associated with surgical recovery. Without adequate nutrition, muscle wasting, immune dysfunction, and declining visceral protein status are observed. While it would seem intuitive that early nutritional intervention is warranted for most patients, the literature to date suggests that early parenteral nutrition (PN) does not improve clinical outcomes. An exception may be for severely malnourished patients if NS is provided for at least seven days preoperatively. Such patients should be identified early through nutritional assessment. Early enteral nutrition (EN) may potentially improve patient outcomes compared with PN, but there are insufficient data to confirm this. High-dose parenteral glutamine may reduce infectious complications and the length of hospitalization for surgical patients, but a significant reduction in mortality has not been observed. Early EN with immune-enhancing formulas appears promising for general surgery patients. However, their use in the critically ill surgical patient is not unanimously supported, and some studies suggest potential harm. CONCLUSION: Surprisingly little evidence is available to support a significant impact of early NS on postoperative clinical outcomes. PMID- 15119574 TI - Model of effect of magnesium prophylaxis on frequency of torsades de pointes in ibutilide-treated patients. PMID- 15119575 TI - Use of Chinese herbal products in Oakland and San Francisco Chinatowns. PMID- 15119576 TI - Current and future considerations for the new classes of biologicals. AB - PURPOSE: Key structural features of biologicals and their development are explained, and the fundamental distinctions between biological and chemical drugs in terms of their discovery, scale-up from research to commercial quantities, quality control, regulatory requirements, and potential for generic substitution are discussed. SUMMARY: Recent advances in biotechnology have accelerated the introduction of biological protein drugs into the marketplace, offering new treatment options and challenges for pharmacists. Because these drugs are produced in living systems and are structurally complex, they are more difficult to manufacture, purify, and evaluate than are traditional chemical drugs. The production of recombinant-DNA-based protein and monoclonal antibody drugs is explained, and the strengths and limitations in selecting one or another host system (i.e., bacteria, yeast, or mammalian cells) for making a given biological drug are explored. Subtle variations in production methods can lead to significant differences in product volume, potential viral or bacterial contamination, bioactivity, and toxicity. Like manufacturers, federal regulators face difficult new challenges because of the structural complexity and in vivo synthesis of biologicals. Pharmacists and regulators alike must determine when and if therapeutic interchange is relevant to biologicals. Because biologicals are so difficult to manufacture and test, noninnovator biologicals must be subject to more oversight than traditional generic drugs. CONCLUSION: Biologicals are complex agents whose production and properties present many considerations that are not associated with traditional chemical drugs. PMID- 15119577 TI - Access to essential drugs in developing countries: a lost battle? PMID- 15119578 TI - Frequency of nausea and vomiting after chemotherapy. PMID- 15119579 TI - Pharmacist involvement in CPOE. PMID- 15119580 TI - Lack of association between fexofenadine and rhabdomyolysis. PMID- 15119581 TI - The influence of storage time and cutting speed on microtensile bond strength. AB - PURPOSE: The aim of this study was to verify the influence of the storage time and the cutting speed during specimen preparation on the bond strength of a single-bottle adhesive to dentin. MATERIALS AND METHODS: A flat dentin surface was exposed in 36 human third molars. The adhesive system (Single Bond) was applied according to the manufacturer's instructions, and composite resin crowns (Z250) were constructed incrementally. Specimens were stored for 10 min, 24 h, or 1 week in distilled water at 37 degrees C before being longitudinally sectioned in both the "x" and "y" directions at different cutting speeds (0.5, 1.6, and 2.6 m/s) to obtain sticks with a cross-sectional area of approximately 0.8 mm2. The specimens were tested in a tensile load machine (0.5 mm/min) and the fracture mode analyzed. A two-way ANOVA with storage time (3 levels) and cutting speed (3 levels) as factors was used to compare the mean microtensile bond strengths. RESULTS: Highly significant main effects and interaction (p < 0.0001) were detected. The highest mean bond strength was obtained with a storage time of 1 week and cutting speed of 2.6 m/s. The lowest mean was found when the specimens were prepared immediately after composite resin placement and sliced at 0.5 m/s. CONCLUSION: Both the storage time and the cutting speed may affect the bond strength results. Therefore, these variables must be controlled in microtensile bond strength tests. PMID- 15119582 TI - Effects of pre- and post-bonding hydration on bond strength to dentin. AB - PURPOSE: This study tested the hypothesis that the hydration condition of the tooth during bonding or storage may affect the bond strength of a single-step, self-etching adhesive to dentin. MATERIALS AND METHODS: Twelve extracted human third molars were transversally sectioned to expose flat, mid-dentin surfaces. Six teeth were dehydrated in ascending ethanol concentrations for several days while 6 teeth were kept hydrated in distilled water. The teeth were bonded with the single-step, self-etching adhesive One-Up Bond F (Tokuyama, Japan) according to directions. Resin composite buildups were constructed incrementally with Z 250 (3M ESPE). The bonded hydrated and dehydrated teeth were then divided into 2 subgroups and stored either in distilled water or in a dry condition for 24 h before being prepared for the microtensile bond strength test. Bonded beams of approximately 0.8 mm2 were tested in tension at 0.5 mm/min in a testing machine. Separate dentin disks were bonded with the adhesive and prepared for transmission electron microscopy (TEM). RESULTS: Bond strengths of specimens stored in water were: 25.0 +/- 10.6 and 18.9 +/- 9.1 MPa for hydrated and dehydrated teeth, respectively. Bond strengths of specimens stored dry were significantly higher (p < 0.001): 46.1 +/- 21.4 and 40.9 +/- 19.1, for hydrated and dehydrated teeth, respectively. Two-way ANOVA showed that previous hydration condition of the teeth had no effect on the bond strength, regardless of the storage condition (p > 0.05). However, storage of the bonded teeth in water caused significant reductions in the bond strength, regardless of the previous hydration condition of the teeth. TEM showed massive silver nitrate impregnation in the adhesive layer of water-stored specimens. CONCLUSIONS: Postbonding water exposure had a highlysignificant effect on bond strength of the single-step, self-etching adhesive. PMID- 15119583 TI - Bonding to cervical sclerotic dentin: effect of acid etching time. AB - PURPOSE: The aim of this in vitro study was to evaluate the microtensile bond strengths (MTBS) of two total-etch one-bottle adhesive systems to natural cervical lesions with 2 different etching times (15 s vs 30 s). MATERIALS AND METHODS: Thirty-two human canines and premolars with saucer-shaped noncarious cervical defects were cleaned and randomly assigned to 4 groups. The adhesive systems - Single Bond (SB; 3M ESPE) or OptiBond Solo Plus (OP; Kerr) - were applied on cervical dentin after acid etching with the proprietary acid gel (35% or 37.5% phosphoric acid) for 15 s (manufacturer's instruction) or for 30 s. The respective hybrid composite was inserted in three increments and light cured. After 24 h water storage, the specimens were cut perpendicularly with a low-speed diamond saw (Isomet) to obtain slices with a cross section of ca 0.8 mm. After that, the slices were trimmed with a diamond bur to obtain a surface area of 0.7 +/- 0.05 mm2 (n = 12). MTBS was measured in a Bencor device with an Instron machine at a crosshead speed of 0.5 mm/min. The data were subjected to two-way ANOVA and Tukey's LSD post-hoc test (p < 0.05). RESULTS: MTBS are given in MPa (mean +/- SD), where means with the same superscript letter are not statistically different at p < 0.05: OP15 = 30.9 +/- 8.8ab, SB15 = 25.6 +/- 9.6bc, OP30 = 19.0 +/- 4.8c, SB30 = 35.9 +/- 11.0a. When data were pooled for "adhesive system", SB resulted in statistically higher bond strengths than OP (p < 0.043). "Acid etching time" had no significant effect (p < 0.766), but the interaction of the two main factors yielded significant differences (p < 0.0001). Extending etching time to 30 s resulted in a more predictable bond to noncarious cervical sclerotic dentin with SB, but resulted in lower MTBS with OP. CONCLUSION: Composite bonding to cervical sclerotic noncarious dentin may depend on acid demineralization capacity and bonding system. Knowing the etching demineralization rates of the commercially available acid etching seems to be an important factor for selecting the best acid-etching time of cervical sclerotic dentin. PMID- 15119584 TI - Marginal integrity of self- and total-etching adhesives in two different application protocols. AB - PURPOSE: The aim of this study was to evaluate the marginal integrity of four self-etching adhesives (Prompt L-Pop III, Resulcin Aqua Prime & MonoBond N, AC Bond, AC Bond + Desensitizer) and three total-etch adhesives (Opti-Bond FL, Excite, Gluma Comfort Bond) in two different application protocols. MATERIALS AND METHODS: Standardized MOD cavities (n = 40) were prepared in 280 extracted human teeth (one cervical margin in cementum, one in enamel). In one application protocol, the adhesive was applied prior to placement of the Tofflemire matrix (n = 20); in the other, the metal matrix was placed first (n = 20). Light curing was carried out from the occlusal aspect. After thermocycling (5000 times, 5 degrees C and 55 degrees C), specimens were immersed in 2% methylene blue for 10 s. Then the median percentages of dye-penetrated margins (surface analysis) were determined separately for the cervical cementum, cervical enamel, and lateral enamel margins of the proximal boxes. RESULTS: At the cervical cementum margins, statistically significantly worse results were obtained for all adhesives when the matrix system was placed prior to application of the adhesive. In the cervical enamel margins, OptiBond FL, Gluma Comfort Bond, and Prompt L-Pop III performed significantly better when the adhesive was applied prior to the matrix. On lateral enamel margins, only OptiBond FL and Resulcin Aqua Prime showed significantly less dye penetration if the matrix was placed first. CONCLUSION: Most of the total-etch materials produced marginal integrity significantly superior to that of the majority of the self-etching materials. Due to a significant influence of the application protocol on marginal integrity, it is strongly recommended to apply the adhesive system before placement of the matrix. PMID- 15119585 TI - Effects of dentin collagen removal on microleakage of bonded restorations. AB - PURPOSE: To evaluate the effects of removing dentin collagen exposed by acid etching on the microleakage of bonded restorations with and without flowable composite application and submitted to thermocycling and long-term water storage. MATERIALS AND METHODS: Standardized Class V cavities were prepared in 180 bovine incisors. They were randomly assigned to three groups according to the adhesive technique used. Conventional group (C): Single Bond was applied according to the manufacturer's instructions and Z250 composite was placed. Hypochlorite group (H): After acid etching a 10% NaOCl aqueous solution was applied for 1 min, then Single Bond and Z250 were applied. Hypochlorite and Flowable group (HF): Similar to group H, but following adhesive application, a thin layer of Natural Flow flowable composite was applied before the Z250. Each group was divided into three subgroups (0, 6, 12), which remained immersed in distilled water for 24 h or 6 or 12 months and underwent 500, 1500 or 2500 thermal cycles, respectively. At the end of each storage time, the specimes were stained with silver nitrate, decalcified, immersed in methyl salicylate for clearing and observed under a stereomicroscope to determine microleakage (scores 0 to 4). The data were analyzed using the Kruskal-Wallis and the Multiple Comparison Tests (alpha = 0.05). RESULTS: After 12 months, every group showed significant increases in microleakage. There was no significant difference between Groups H and HF for the three different periods of time, but they showed statistically less microleakage than Group C. CONCLUSION: The removal of dentinal collagen reduced the marginal microleakage when compared to the conventional technique. The use of the flowable composite did not produce significant effects. No technique was completely effective in preventing microleakage. PMID- 15119586 TI - Microleakage of a pit-and-fissure sealant: effect of air-abrasion compared with classical enamel preparations. AB - PURPOSE: The aim of this study was to assess the microleakage of a pit-and fissure sealant after classical enamel preparations (prophylaxis followed by acid etching alone and mechanical widening with bur) and after air abrasion. MATERIALS AND METHODS: Ninety noncarious freshly extracted third molars were randomly assigned to three groups of 30. In each group, the mesial halves of the fissures were treated with air abrasion and conditioned with acid etching for 15 s. In group 1, the distal halves were treated with acid etching alone. In group 2, the distal halves of the fissures were widened mechanically with a bur and etched for 15 s. In group 3, the distal halves were prepared with air abrasion alone. Then the sealant (Clinpro) was applied on the occlusal fissures of all teeth, according to the manufacturer's recommendations. The teeth were thermocycled and the samples placed in a 1% methylene blue dye solution. The teeth showing microleakage and the means of infiltration were assessed with an image analysis system. RESULTS: The microleakage of sealants prepared with air abrasion alone displayed significantly greater microleakage (80%) (p < 0.0001) than the ones placed after prophylaxis and etching (13.33%), bur and etching (20%), or air abrasion and etching (22.2%). The samples prepared with air abrasion alone also showed the highest mean microleakage (1.14 +/- 1.4 mm) when compared to samples receiving prophylaxis and acid etching (0.12 +/- 0.41), bur and acid etching (0.08 +/- 0.26), or air abrasion and acid etching (0.09 +/- 0.4) (p < 0.05). No significant difference was noted between the three types of enamel preparation when etching was performed. CONCLUSION: Air-abrasion treatment does not eliminate the need for etching the enamel surface before applying the sealant. PMID- 15119587 TI - In vitro marginal adaptation of high-viscosity resin composite restorations bonded to dentin cavities. AB - PURPOSE: The aim of this study was to evaluate the marginal adaptation of high viscosity resin composite restoratives bonded to dentin in a cylindrical cavity model. MATERIALS AND METHODS: The buccal enamel of 64 human premolars was removed and cylindrical cavities 3 mm in diameter and 1.3 mm in depth were prepared on each dentin surface. The cavities were divided into 8 groups of 8 cavities each and restored according to the manufacturers' instructions with the following adhesive/composite systems: Bond 1/Alert, Stae/Glacier, OptiBond Solo/Prodigy Condensable, One-Step/Pyramid, Solidbond/Solitaire, Prime&Bond NT/Surefil, One Coat Bond/Synergy, and Scotchbond 1/Z250. The composite surfaces were pressed against mylar strips, covered with cover slips, and photopolymerized in a single increment for 40 s. The restorations were polished with wet SiC papers of 320 to 1000 grit size to expose dentin margins. The marginal adaptation was evaluated immediately after photopolymerization and again after 1 week of storage in water at 37 +/- 1 degrees C. Evaluation was performed under a metallographic microscope at 200X magnification by recording the frequency of gap-free restorations (GF), the percentage length of the debonded margins relative to the cavity periphery (DM), the width of the maximum marginal gap (MG), and the marginal index (MI = MG x DM / 100). The results were statistically analyzed with one-way ANOVA and the Mann-Whitney U-test at alpha = 0.05. RESULTS: No incidence of gaps was found in 62.5% of One Coat Bond/Synergy and 37.5% of OptiBond Solo/Prodigy Condensable restorations. All the other restorative systems exhibited restorations with gaps. One Coat Bond/Synergy, Scotchbond 1/Z250, and OptiBond Solo/Prodigy Condensable were the groups with the lowest DM values, while Stae/Glacier showed the highest DM values. One Coat Bond/Synergy and OptiBond Solo/Prodigy Condensable revealed the lowest MI values and Stae/Glacier the highest. No statistically significant differences were recorded between immediate evaluation and evaluation after 1 week of water storage. CONCLUSION: The results showed that all the adhesive/composite systems examined failed to achieve gap-free margins with dentin. The restorative systems One Coat Bond/Synergy, Optibond Solo/Prodigy Condensable, and Scotchbond 1/Z250 presented the best marginal adaptation of all the systems tested. No improvement was recorded on the quality of marginal adaptation following 1 week of water storage. PMID- 15119588 TI - Self-etching adhesives improve the shear bond strength of a resin-modified glass ionomer cement to dentin. AB - PURPOSE: Despite the improvements of resin-modified glass-ionomer cements, the dentin bond strength of these materials remains inferior to that provided by "bonding system/composite" combinations and limits the clinical use of RMGIC. The objective of this in vitro study was to investigate the effect of new self etching bonding systems on the adhesion to dentin of Fuji II LC. MATERIALS AND METHODS: Seven recent self-etching systems were studied (Clearfil SE Bond = CSE, Optibond Solo Plus Self-Etching Primer = OSP, One Up Bond F = OUP, Mac Bond II = MB2, Xeno III = XB3, ADPER Prompt-L-Pop = ADP, Unifil Bond = UB) and compared with Cavity Conditioner (C). After the bonding procedure, a resin-modified glass ionomer cylinder (Fuji II LC) was bonded to the surface using a Teflon mold (diameter: 3 mm; height: 5 mm). The different specimens were tested in a shear bond mode (0.5 mm/min). Statistical analyses were performed with one-way ANOVA and Fischer PLSD test (p < 0.05). RESULTS: The one-way ANOVA revealed a significant influence of the surface treatment parameter (p < 0.0001). The lowest shear bond strength values were obtained with Cavity Conditioner. The highest values were obtained with MB2. The multiple comparisons revealed that ADP = OBP = XB3 = OUB = UB = CSE and that UB = CSE = MB2. CONCLUSION: The application of the 7 self-etching adhesives tested in this study improves the dentin shear bond strengths of Fuji II LC. This increase depended on the adhesive system and ranged from 50% for ADP up to 130% for MB2. PMID- 15119589 TI - Glutaraldehyde in bonding systems disinfects dentin in vitro. AB - PURPOSE: To study the dentin disinfecting capacity of glutaraldehyde-containing bonding agents. MATERIALS AND METHODS: Uniformly sized bovine dentin blocks infected with Enterococcus faecalis ATCC 29212 were treated according to manufacturer's instructions with a three-step bonding system (Syntac Classic), its single components, or Gluma Primer. Bacterial growth from dentinal filings obtained immediately after application of the bonding agents was assessed at different dentin depths. Minimal inhibitory concentration and minimal bactericidal concentration of glutaraldehyde against E. faecalis were determined. RESULTS: Both Gluma Primer and Syntac Adhesive disinfected dentin, while the other components failed to do so. This was explained by the high antibacterial efficacy of glutaraldehyde even at low concentrations. Applied in consecutive order, the Syntac bonding system disinfected dentin blocks equally well with or without previous application of Gluma Primer. CONCLUSION: Under the conditions of the current study, bonding agents containing glutaraldehyde appear to be good dentin disinfectants compatible with bonding systems. However, their effect on pulp tissue awaits further clarification. PMID- 15119590 TI - A prospective ten-year clinical trial of porcelain veneers. AB - PURPOSE: The aim of this in vivo study was to evaluate the clinical performance of porcelain veneers after 5 and 10 years of clinical service. MATERIALS AND METHODS: A single operator placed porcelain laminates on 87 maxillary anterior teeth in 25 patients. All restorations were recalled at 5 years and 93% of the restorations at 10 years. Clinical performance was assessed in terms of esthetics, marginal integrity, retention, clinical microleakage, caries recurrence, fracture, vitality, and patient satisfaction. Failures were recorded either as "clinically unacceptable but repairable" or as "clinically unacceptable with replacement needed". RESULTS: Porcelain veneers maintained their esthetic appearance after 10 years of clinical service. None of the veneers were lost. The percentage of restorations that remained "clinically acceptable" (without need for intervention) significantly decreased from an average of 92% (95 CI: 90% to 94%) at 5 years to 64% (95 CI: 51% to 77%) at 10 years. Fractures of porcelain (11%) and large marginal defects (20%) were the main reason for failure. Marginal defects were especially noticed at locations where the veneer ended in an existing composite filling. At such vulnerable locations, severe marginal discoloration (19%) and caries recurrence (10%) were frequently observed. Most of the restorations that present one or more "clinically unacceptable" problems (28%) were repairable. Only 4% of the restorations needed to be replaced at the 10-year recall. CONCLUSION: It was concluded that labial porcelain veneers represent a reliable, effective procedure for conservative treatment of unesthetic anterior teeth. Occlusion, preparation design, presence of composite fillings, and the adhesive used to bond veneers to tooth substrate are covariables that contribute to the clinical outcome of these restorations in the long-term. PMID- 15119591 TI - Photoaffinity scanning in the mapping of the peptide receptor interface of class II G protein-coupled receptors. AB - The family of G protein-coupled receptors constitutes about 50% of the therapeutic drug targets used in clinical medicine today, although the mechanisms of ligand binding, activation and signal transduction for G protein-coupled receptors are not yet well defined. This review discusses ongoing research using the photoaffinity scanning method to map the bimolecular interface between class II G protein-coupled receptors and their ligands. Furthermore the available computer model of class II peptide ligand docking into the receptor, based on the positional constraints imposed by the photoaffinity scanning analyses, will be discussed briefly. The ultimate goal of these efforts is to understand the molecular basis of receptor binding and therefore to generate a template for rational drug design. PMID- 15119592 TI - HP(2-9)-magainin 2(1-12), a synthetic hybrid peptide, exerts its antifungal effect on Candida albicans by damaging the plasma membrane. AB - In our previous study, HP(2-9)-MA(1-12), HP-MA for short, a hybrid peptide incorporating residues 2-9 of Helicobacter pylori ribosomal protein L1 (HP) and residues 1-12 of magainin 2 (MA) was shown to have strong antibacterial activity. In this study the antifungal activity of HP-MA was evaluated using various fungi, and it was shown that the activity was increased when compared with the parent peptides. In order to investigate the fungicidal mechanism(s) of HP-MA its action against fungal cell membranes was examined by the potassium-release test, which showed that HP-MA caused an increase in the amount of K+ released from the cells. Furthermore, HP-MA induced significant morphological changes. These facts suggested that the fungicidal effect of HP-MA involves damaging the fungal cell membranes. CD investigators suggested that the alpha-helical structure of these peptides plays an important role in their antibiotic effect, but that alpha helicity is less directly correlated with the enhanced antibiotic activity of the hybrid. PMID- 15119593 TI - Characterizing bathocuproine self-association and subsequent binding to Alzheimer's disease amyloid beta-peptide by NMR. AB - Aggregated amyloid beta-peptide (A beta) is the primary constituent of the extracellular plaques and perivascular amyloid deposits associated with Alzheimer's disease (AD). Deposition of the cerebral amyloid plaques is thought to be central to the disease progression. One such molecule that has previously been shown to 'dissolve' deposited amyloid in post-mortem brain tissue is bathocuproine (BC). In this paper 1H NMR chemical shift analysis and pulsed field gradient NMR diffusion measurements were used to study BC self-association and subsequent binding to A beta. The results show that BC undergoes self-association as its concentration increases. The association constant of BC dimerization, Ka, was estimated to be 0.64 mM(-1) at 25 degrees C from 1H chemical shift analysis. It was also found that dimerization of BC appeared to be essential for its binding to A beta. From the self-association constant of BC, Ka, the fraction of dimeric BC in the complex was obtained and the dissociation constant, Kd, of BC bound to A beta40 peptide was then determined to be approximately 1 mM. PMID- 15119594 TI - On-resin head-to-tail cyclization of cyclotetrapeptides: optimization of crucial parameters. AB - Cyclotetrapeptides are constrained cyclic peptides whose synthesis is considered a difficult task. A methodology based on on-resin head-to-tail cyclization by anchoring the side chain of a trifunctional amino acid was investigated. A series of model cyclotetrapeptides containing the RGD sequence cyclo(Xaa-Arg-Gly-Asp) (Xaa = Ala, Phe, Phg, D-Ala, D-Phe, D-Phg) was synthesized with no cyclodimerization by-products. An evaluation and optimization study of all of the parameters directly involved in the ring closure was performed. PMID- 15119595 TI - Pesticide residues in food--acute dietary exposure. AB - Consumer risk assessment is a crucial step in the regulatory approval of pesticide use on food crops. Recently, an additional hurdle has been added to the formal consumer risk assessment process with the introduction of short-term intake or exposure assessment and a comparable short-term toxicity reference, the acute reference dose. Exposure to residues during one meal or over one day is important for short-term or acute intake. Exposure in the short term can be substantially higher than average because the consumption of a food on a single occasion can be very large compared with typical long-term or mean consumption and the food may have a much larger residue than average. Furthermore, the residue level in a single unit of a fruit or vegetable may be higher by a factor (defined as the variability factor, which we have shown to be typically x3 for the 97.5th percentile unit) than the average residue in the lot. Available marketplace data and supervised residue trial data are examined in an investigation of the variability of residues in units of fruit and vegetables. A method is described for estimating the 97.5th percentile value from sets of unit residue data. Variability appears to be generally independent of the pesticide, the crop, crop unit size and the residue level. The deposition of pesticide on the individual unit during application is probably the most significant factor. The diets used in the calculations ideally come from individual and household surveys with enough consumers of each specific food to determine large portion sizes. The diets should distinguish the different forms of a food consumed, eg canned, frozen or fresh, because the residue levels associated with the different forms may be quite different. Dietary intakes may be calculated by a deterministic method or a probabilistic method. In the deterministic method the intake is estimated with the assumptions of large portion consumption of a 'high residue' food (high residue in the sense that the pesticide was used at the highest recommended label rate, the crop was harvested at the smallest interval after treatment and the residue in the edible portion was the highest found in any of the supervised trials in line with these use conditions). The deterministic calculation also includes a variability factor for those foods consumed as units (eg apples, carrots) to allow for the elevated residue in some single units which may not be seen in composited samples. In the probabilistic method the distribution of dietary consumption and the distribution of possible residues are combined in repeated probabilistic calculations to yield a distribution of possible residue intakes. Additional information such as percentage commodity treated and combination of residues from multiple commodities may be incorporated into probabilistic calculations. The IUPAC Advisory Committee on Crop Protection Chemistry has made 11 recommendations relating to acute dietary exposure. PMID- 15119596 TI - Paraquat and sustainable agriculture. AB - Sustainable agriculture is essential for man's survival, especially given our rapidly increasing population. Expansion of agriculture into remaining areas of natural vegetation is undesirable, as this would reduce biodiversity on the planet. Maintaining or indeed improving crop yields on existing farmed land, whether on a smallholder scale or on larger farms, is thus necessary. One of the limiting factors is often weed control; biological control of weeds is generally of limited use and mechanical control is either often difficult with machinery or very laborious by hand. Thus the use of herbicides has become very important. Minimum cultivation can also be important, as it reduces the power required to work the soil, limits erosion and helps to maintain the organic matter content of the soil. This last aspect helps preserve both the structure of soil and its populations of organisms, and also sustains the Earth's soil as a massive sink for carbon, an important consideration in the light of global warming. The introduction of the bipyridinium herbicide paraquat in the early 1960s greatly facilitated weed control in many crops. Paraquat has the unusual property of being active only by direct spray onto plants and not by uptake from soil in which strong binding deactivates it. Together with its rapid action in light in killing green plant tissue, such properties allow paraquat to be used in many crops, including those grown by low-tillage methods. This paper reviews the ways in which agricultural systems have been and are being developed to make use of these properties, and provides a risk/benefit analysis of the world-wide use of paraquat over nearly 40 years. PMID- 15119597 TI - A mini-bag technique for evaluation of fungicide effects on Trichoderma spp in mushroom compost. AB - An in vivo technique was developed to observe colonisation of mushroom compost by Trichoderma spp. Isolates of T. harzianum (Th2), T. harzianum (Th1), T. koningii (Tk) and T. viride (Tv) were artificially introduced into compost using a mini bag system. Wheat grains, colonised by Trichoderma spp, were placed centrally on a layer of compost at the bottom of 1-litre polythene bags which were then filled with 350 g of spawned or un-spawned compost, and partially sealed. After 14 and 21 days incubation at 27 degrees C, the bags were assessed for recovery of Trichoderma from middle and top zones using a needle stab re-isolation technique and a visual colonisation scoring system. Visible green mould contamination, similar to that observed in practice, developed within 21 days. The visual colonisation scoring was reliably related to the re-isolation success. In this evaluation, Trichoderma spp showed considerable differences in their relative abilities to colonise spawned and un-spawned compost, with Th2 isolates being consistently superior to the other isolates of Th1, Tk and Tv in colonising spawned compost. This technique was employed to evaluate the effects of fungicides on the colonisation of mushroom compost by three Trichoderma spp: Th2, Th1 and Tk, using 1-litre and 5-litre mini-bag systems. Aqueous suspensions of benomyl, carbendazim, thiabendazole, prochloraz and prochloraz+carbendazim incorporated into the compost at 50 mg litre(-1), or applied to spawn at 50 mg kg(-1), reduced the colonisation by Trichoderma spp. Prochloraz and prochloraz+carbendazim were superior to benomyl, carbendazim or thiabendazole in reducing compost colonisation by Th2, Th1 and Tk, with Th2 being the most persistent type, capable of colonising treated compost in the presence of all five fungicides. The prochloraz+carbendazim mixture, not normally used in mushroom production, was equal to or better than prochloraz alone. The incidence of green mould colonisation by Th2 was as extensive in the 5-litre compost bags as in the 1-litre bags, but colonisation by Th1 and Tk was more apparent in the 5 litre bags. The in vivo mini-bag evaluations using wheat grain Trichoderma inoculum and needle stab re-isolation procedures proved an efficient method for studying colonisation and screening for effectiveness of fungicides applied to mushroom compost or spawn. PMID- 15119599 TI - Fumigant combinations for Cyperus esculentum L control. AB - The phase-out of methyl bromide as a soil fumigant has stimulated research into the use of other soil fumigants for weed control. Methyl bromide, methyl iodide, propargyl bromide, 1,3-dichloropropene (1,3-D) and metam-sodium were tested alone and in combination with chloropicrin in laboratory experiments to determine their efficacy against Cyperus esculentus L (yellow nutsedge) tubers. Propargyl bromide and metam-sodium were the most efficacious fumigants tested, with EC50 values of 3.7 and 6.5 microM, respectively. The relative potencies of methyl iodide and chloropicrin were not significantly different but were 2.6 and 2.9 times more potent than methyl bromide, respectively. The EC50 values for all fumigants other than 1,3-D were significantly lower than that of methyl bromide. Combining each fumigant with 17% chloropicrin resulted in a synergistic interaction. The greatest increase in potency between the expected result and the actual result was a relative potency of 3.8 with the methyl bromide/chloropicrin combination. The smallest increase in efficacy was with propargyl bromide and chloropicrin, with a relative potency of 1.5. There was no significant difference between the EC50 values of methyl bromide/chloropicrin and methyl iodide/chloropicrin combinations. Combining 1,3-D with 17% chloropicrin resulted in an EC50 value for C. esculentus control similar to that of methyl iodide applied alone. PMID- 15119598 TI - Pyrethroid tolerance in Culex pipiens pipiens var molestus from Marin County, California. AB - In May 2001 a sample of Culex pipiens pipiens variety molestus Forskal from Marin County, California, collected as larvae and reared to adults, was found to show reduced resmethrin and permethrin knock-down responses in bottle bioassays relative to a standard susceptible Cx. pipiens quinquefasciatus Say colony (CQ1). Larval susceptibility tests, using CQ1 as standard susceptible, indicated that the Marin mosquitoes had LC50 resistance ratios of 18.3 for permethrin, 12 for deltamethrin and 3.3 for pyrethrum. A colony of Marin was established and rapidly developed higher levels of resistance in a few generations after exposure to permethrin as larvae. These selected larvae were shown to cross-resist to lambda cyhalothrin as well as to DDT. However, adult knock-down time in the presence of permethrin, resmethrin and pyrethrum was not increased after increase in tolerance to pyrethroids as larvae. Partial and almost complete reversion to susceptibility as larvae was achieved with S, S, S-tributylphosphorotrithioate and piperonyl butoxide (PBO), respectively, suggesting the presence of carboxylesterase and P450 monooxygenase mediated resistance. Insensitive target site resistance (kdr) was also detected in some Marin mosquitoes by use of an existing PCR-based diagnostic assay designed for Cx. p. pipiens L mosquitoes. Carboxylesterase mediated resistance was supported by use of newly synthesized novel pyrethroid-selective substrates in activity assays. Bottle bioassays gave underestimates of the levels of tolerance to pyrethroids of Marin mosquitoes when compared with mortality rates in field trials using registered pyrethroid adulticides with and without PBO. This study represents the first report of resistance to pyrethroids in a feral population of a mosquito species in the USA. PMID- 15119600 TI - Efficacy of insecticide mixtures against larvae of Culex quinquefasciatus (Say) (Diptera: Culicidae) resistant to pyrethroids and carbamates. AB - The efficacy of insecticide mixtures of permethrin (pyrethroid) and propoxur (carbamate) was tested by larval bioassays on two strains of Culex quinquefasciatus (Say), one resistant to pyrethroids and the other resistant to carbamates. The method consisted in combining one insecticide at the highest concentration causing no mortality (LC0) with increasing concentrations of the second one. The concentration-mortality regression lines were determined for permethrin and propoxur alone and in combination, and synergism ratios (SR) were calculated in order to determine the magnitude of an increase or decrease in efficacy with use of the mixtures. With the pyrethroid-resistant strain (BK-PER), the results showed that propoxur at LC0 significantly enhanced the insecticidal activity of permethrin (SR50 = 1.54), especially on the upper range of the concentration-mortality regression. Conversely, when permethrin at LC0 was tested with propoxur against the carbamate resistant strain (R-LAB), an antagonistic effect was observed (SR50 = 0.67). With the BK-PER strain, an increased oxidative detoxification (MFO) appeared to be the main mechanism responsible for the synergistic interaction. Nevertheless, antagonism in the R-LAB strain is probably due to a physiological perturbation implying different target sites for pyrethroid (ie sodium channel) and carbamate insecticides [ie acetylcholinesterase (EC 3.3.3.7) and choline acetyltransferase (EC 2.3.1.6)]. PMID- 15119601 TI - Effects of sub-lethal and lethal doses of lambda-cyhalothrin on oviposition experience and host-searching behaviour of a parasitic wasp, Aphidius ervi. AB - In many parasitoid species, the recognition of chemical signals is essential to find specific hosts. This function is often impaired by exposure to insecticides that are usually neurotoxic. The behaviour of the Hymenopterous parasitoid Aphidius ervi (Haliday) (Hymenoptera: Aphidiinae) after surviving low doses of the pyrethroid lambda-cyhalothrin was examined in laboratory conditions. The host aphid was Myzus persicae (Sulzer) (Homoptera: Aphididae) on oilseed rape. Parasitoid females were exposed by contact with dry residues of the active ingredient at a lethal dose, LD20, and a sub-lethal dose, LD0.1. In a four-armed olfactometer, untreated and inexperienced females were attracted by the odour of M. persicae-infested plants and previous oviposition experience increased the duration of the attraction response. The response of inexperienced females decreased after an exposure to LD0.1 but not to LD20. No effect was observed when females had an oviposition experience prior to the olfactometer test. The oviposition activity was significantly decreased in the LD20-treated group but not in the LD0.1-treated one. All effects disappeared within 24h. Our work shows that orientation and oviposition behaviours may be impaired by low doses of lambda-cyhalothrin, depending on the dose, the parasitoid experience and the type of behaviour. PMID- 15119602 TI - Diffusion and emissions of 1,3-dichloro propene in Florida sandy soil in microplots affected by soil moisture, organic matter, and plastic film. AB - The main objective of this study was to determine the influence of soil moisture, organic matter amendment and plastic cover (a virtually impermeable film, VIF) on diffusion and emissions of (Z)- and (E)-1,3-dichloropropene (1,3-D) in microplots of Florida sandy soil (Arredondo fine sand). Upward diffusion of the two isomers in the Arredondo soil without a plastic cover was greatly influenced by soil water content and (Z)-1,3-D diffused faster than (E)-1,3-D. In less than 5 h after 1,3-D injection to 30 cm depth, (Z)- and (E)-1,3-D in air dry soil had diffused to a 10 cm depth, whereas diffusion for the two isomers was negligible in near-water-saturated soil, even 101 h after injection. The diffusion rate of (Z)- and (E)-1,3-D in near-field-capacity soil was between the rates in the two water regimes. Yard waste compost (YWC) amendment greatly reduced diffusion of (Z)- and (E)-1,3-D, even in air-dry soil. Although upward diffusion of (Z)- and (E)-1,3-D in soil with VIF cover was slightly less than in the corresponding bare soil; the cover promoted retention of vapors of the two isomers in soil pore air in the shallow subsurface. More (Z)-1,3-D vapor was found initially in soil pore air than (E)-1,3-D although the difference declined thereafter. As a result of rapid upward movement in air-dry bare soil, (Z)- and (E)-1,3-D were rapidly volatilized into the atmosphere, but emissions from the near-water-saturated soil were minimal. Virtually impermeable film and YWC amendment retarded emissions. This study indicated that adequate soil water in this sandy soil is needed to prevent rapid emissions, but excess soil water slows diffusion of (Z)- and (E) 1,3-D. Thus, management for optimum water in soil is critical for pesticidal efficacy and the environment. PMID- 15119603 TI - Synthesis and structure-activity study of fungicidal anilinopyrimidines leading to mepanipyrim (KIF-3535) as an anti-Botrytis agent. AB - A series of 2-anilinopyrimidines was prepared and their fungicidal activities against Botrytis cinerea Pers were examined. The activity fell sharply with any substitution on the anilinobenzene ring. Substituents at the 5-position of the pyrimidine ring greatly reduced the activity. Substituents such as chloro, methoxy, methylamino, methyl or 1-propynyl were well tolerated at the 4- and 6 positions of the pyrimidine ring. Among these substituents, the combination of methyl and 1-propynyl groups was the most favourable. 2-Anilino-4-methyl-6-(1 propynyl)pyrimidine (KIF-3535), which showed excellent activity and no significant phytotoxicity, was finally selected for development and has been given the common name mepanipyrim. PMID- 15119604 TI - Esterase-mediated tolerance to a formulation of the organophosphate insecticide monocrotophos in the entomopathogenic fungus, Beauveria bassiana (Balsamo) Vuill: a promising biopesticide. AB - The use of biopesticides compatible with chemical pesticides is recommended in pest management as an effective and ecologically sound strategy. The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuill, which is used as a biopesticide, was found to exhibit and lose tolerance to two organophosphorus insecticides widely used in Indian fields. The pattern of response is similar to the esterase-mediated organophosphate tolerance in aphids achieved through esterase gene duplication. Therefore the role of esterase in the tolerance exhibited by isolates of B. bassiana to the organophosphate, monocrotophos, was studied. Both the total soluble protein content and esterase activity were found to increase significantly in B. bassiana cultures that were able to grow in the presence of monocrotophos. With the hitherto established linkage between esterase overproduction and organophosphate tolerance in insects and the observed similarity in response of the insects and B. bassiana to the chemical, it is concluded that tolerance to monocrotophos in B. bassiana may be due to a mechanism similar to that operating in insects. Habituation of the fungus to monocrotophos to initiate expression of esterase gene may help in achieving compatibility between the two. PMID- 15119605 TI - Insecticidal activity of chemical constituents from Aristolochia pubescens against Anticarsia gemmatalis larvae. AB - Acetone and ethanol extracts of the tubercula and several compounds isolated from Aristolochia pubescens (Willd) were bioassayed on velvetbean caterpillars, Anticarsia gemmatalis (Hubner), for evaluation of the insecticidal activities. Of the extracts subjected to bioassay, the acetone extract showed the highest activity. (-)-Cubebin did not show activity against soybean caterpillars, whereas aristolochic acid and ent-kaur-15-en-17-ol increased the larval period. These compounds, and (+)-eudesmin and (+)-sesamin, reduced the viability of this period, giving rise to malformed adults. These extracts and compounds are therefore potential botanical insecticide agents for the control of velvetbean caterpillars in soybean crops. PMID- 15119606 TI - Temperature sensor based on the power reflected by a Bragg grating in a tapered fiber. AB - We present a temperature sensor based on two chirped gratings made in optical fibers tapered by fusion. One of the gratings has a metallic shielding and acts as sensor element, whereas the second grating provides a reference signal. The sensor is interrogated by measuring the power reflected by the two gratings, and the system has an accuracy of 0.05 degrees C over a linear operation range of more than 10 degrees C that can be adjusted in the fabrication process. PMID- 15119607 TI - Adjustable spiral phase plate. AB - A spiral phase retarder phi(r, theta) = mtheta has been constructed with use of a deformed cracked plexiglass plate. By changing the degree of deformation, the retarder can be adjusted for use at any wavelength, and the value of the phase step 2pim at theta = 2pi can be chosen. PMID- 15119608 TI - Digital speckle pattern interferometry for deformation analysis of inner surfaces of cylindrical specimens. AB - Deformation study of curved engineering and technical surfaces, such as pipes and pressure vessels, has gained much importance in the recent past. Speckle interferometric techniques and their electronic and digital analogs, which are whole field techniques, have been effectively applied for practical nondestructive testing applications over the years. However, little work has been done that discusses the speckle fringe formation with a fruitful theoretical formulation to study deformation analysis of curved surfaces. We propose an extended theory for speckle fringe formation on curved surfaces, which can be applied to the study of curved engineering and technical specimens under various loading conditions such as in-plane, out-of-plane, and out-of-plane shear configurations. Simulated contours are generated by use of finite element models with similar loading conditions, and the data are analyzed and compared with the obtained experimental results. PMID- 15119609 TI - Excitation of surface-plasmon polaritons by use of a zeroth-order Bessel beam. AB - We report an experimental result that shows the excitation of surface plasmon polaritons by use of a zeroth-order Bessel beam. From the viewpoint of energy efficiency, the zeroth order Bessel beam is ideal for the local excitation of surface plasmon polaritons on a metal film. We introduce an optical setup using an axicon element in order to carry out the excitation. PMID- 15119610 TI - Microfluorometric detection of catecholamines with multiphoton-excited fluorescence. AB - We demonstrate sensitive spatially resolved detection of physiological chromophores that emit in the ultraviolet (<330 nm). An atypical laser source (a visible wavelength femtosecond optical parametric oscillator), and an unconventional collection geometry (a lensless detector that detects the forward emitted fluorescence) enable this detection. We report the excitation spectra of the catecholamines dopamine and norepinephrine, together with near-UV emitters serotonin and tryptophan, in the range of 550-595 nm. We estimate the molecular two-photon action cross section of dopamine, norepinephrine, and serotonin to be 1.2 mGM (1 GM, or Goppert Mayor, is equal to 10(-58) m4 s(-1) photon(-1)), 2 mGM, and 43 mGM, respectively, at 560 nm. The sensitivity achieved by this method holds promise for the microscopic imaging of vesicular catecholamines in live cells. PMID- 15119611 TI - Optical phase retrieval by use of first Born- and Rytov-type approximations. AB - The first Born and Rytov approximations of scattering theory are introduced in their less familiar near-field versions. Two algorithms for phase retrieval based on these approximations are then described. It is shown theoretically and by numerical simulations that, despite the differences in their formulation, the two algorithms deliver fairly similar results when used for optical phase retrieval in the near and intermediate fields. The algorithms are applied to derive explicit solutions to four phase-retrieval problems of practical relevance to quantitative phase-contrast imaging and tomography. An example of successful phase reconstruction by use of the Born-type algorithm with an experimental x-ray image is presented. PMID- 15119612 TI - Electromagnetic design of an all-diffractive millimeter-wave imaging system. AB - We present the design and electromagnetic analysis of an all-diffractive millimeter-wave imaging system having a field of view of +/- 15 degrees. This system consists of two 16-level diffractive lenses, with the stop in contact with the first lens. By considering the Seidel aberrations for a diffractive lens and applying the corresponding stop shift formula, we established the expressions of third-order wave aberrations for this system. By setting all primary Seidel aberrations to zero and solving the corresponding system of equations, we obtained two sets of solutions for this two-element all-diffractive system, which totally compensate for all Seidel aberrations. To assess image system performance, we apply the finite-difference time-domain technique and a vector plane-wave spectrum method, in combination, to validate the performance of the system. To reduce the computational cost and thereby enable the complete electromagnetic analysis of the system, a four-step analysis procedure has been developed and applied as an electromagnetic system model. PMID- 15119613 TI - Electronic speckle pattern shearing interferometer with a photopolymer holographic grating. AB - A photopolymer holographic grating is used to produce the two sheared images in an electronic speckle pattern shearing interferometer. A ground glass screen following the grating eliminates unwanted diffraction orders and removes the requirement for the CCD camera to resolve the diffraction grating's pitch. The sheared images on the ground glass are further imaged onto the CCD camera. The fringe pattern contrast was estimated to be above 90%. A validation of the system was done by comparing the theoretical phase difference distribution with the experimental data from the three-point bending test. PMID- 15119614 TI - Removing nonlinearity of a homodyne interferometer by adjusting the gains of its quadrature detector systems. AB - Most homodyne interferometers have a quadrature detector system that includes two polarizing beam splitters that cause nonlinearity of the order of a few nanometers by phase mixing. Detectors should have the same gains to reduce nonlinearity under the assumption that there is no loss in optical components. However, optical components exhibit some loss. We show that nonlinearity can be reduced to an order of 0.01 nm when the detector gains are adjusted by simulation to include the optical characteristics. The compensated nonlinearity is 18 times smaller than that when the four detector gains are set to be equal. PMID- 15119615 TI - Three-dimensional measurement with use of a single camera. AB - A new, to our knowledge, measurement technique that uses a single image of a handheld probe on known points, to measure geometric features of an object, is discussed in detail. A new method is also proposed to calculate three-dimensional coordinates of points on the object relative to a reference. Furthermore, the validity of the proposed measuring system is confirmed by experiments. PMID- 15119616 TI - Experimental techniques using optically enabled ring oscillators. AB - We present a method for characterizing transceiver performance in parallel optical data links. By establishing a bidirectional link between two optically enabled chips, the conditions for a ring oscillator are made possible. We propose this technique as a means of measuring the sensitivity of transceiver latency to controlled variables such as temperature, optical output power, supply voltage, and device misalignment. This method is noninvasive and independent of the packaging, circuit topology, and optical medium used. The technique is demonstrated experimentally, and results are compared with a simulation. PMID- 15119617 TI - Simulation of sea surface wave influence on small target detection with airborne laser depth sounding. AB - A theoretical model for simulation of airborne depth-sounding lidar is presented with the purpose of analyzing the influence from water surface waves on the ability to detect 1-m3 targets placed on the sea bottom. Although water clarity is the main limitation, sea surface waves can significantly affect the detectability. The detection probability for a target at a 9-m depth can be above 90% at 1-m/s wind and below 80% at 6-m/s wind for the same water clarity. The simulation model contains both numerical and analytical components. Simulated data are compared with measured data and give realistic results for bottom depths between 3 and 10 m. PMID- 15119618 TI - Aerosol optical thickness from Brewer spectrophotometers and an investigation into the stray-light effect. AB - The Langley method has been applied to the measurements of direct solar radiation made by Brewer spectrophotometers to obtain the aerosol optical thickness (AOT) of the atmosphere in the ultraviolet-B range. In several cases the AOT increased with wavelength, which raises suspicion about the stray-light effect. To investigate the quality of the AOT measurements and the possibility of stray light, we conducted a campaign by using single- and double-monochromator Brewers. The campaign's results have shown that both Brewers' AOT values are in good agreement and that stray light is not an important effect for AOT at wavelengths above 306 nm. PMID- 15119619 TI - Geometrical-optics solution to light scattering by droxtal ice crystals. AB - We investigate the phase matrices of droxtals at wavelengths of 0.66 and 11 microm by using an improved geometrical-optics method. An efficient method is developed to specify the incident rays and the corresponding impinging points on the particle surface necessary to initialize the ray-tracing computations. At the 0.66-microm wavelength, the optical properties of droxtals are different from those of hexagonal ice crystals. At the 11-microm wavelength, the phase functions for droxtals are essentially featureless because of strong absorption within the particles, except for ripple structures that are caused by the phase interference of the diffracted wave. PMID- 15119620 TI - Properties of aqueous methanesulfonic acid: complex index of refraction and surface tension. AB - The complex index of refraction of aqueous methanesulfonic acid (MSA) at room temperature and at concentrations of 1-70 wt. % is presented. The complex index of refraction is obtained in the entire wave-number region from the mid infrared to the ultraviolet-visible regions. An analysis of the infrared spectra reveals a single absorption band, which makes it possible to identify MSA in aqueous H2SO4. The surface tension of 1-99-wt. % aqueous MSA from room temperature to approximately 263 K, depending on the concentration of MSA, is reported. A polynomial parameterization of the surface tension in terms of acid weight fraction and temperature is presented. PMID- 15119621 TI - Detailed study of an efficient blue laser source by second-harmonic generation in a semimonolithic cavity for the cooling of strontium atoms. AB - We have constructed a blue laser source consisting of an amplified, grating tuned diode laser that is frequency doubled by a KNbO3 crystal in a compact standing wave cavity and produces as much as 200 mW of internal second-harmonic power. We have analyzed the unusual characteristics of this standing wave cavity to clarify the advantages and disadvantages of this configuration as an alternative to a ring cavity for second-harmonic generation. We emphasize its efficiency and stability and the fact that it has an inherent walk-off compensation, similar to twin crystal configurations. We demonstrate its utility for laser cooling and trapping of earth alkalis by stabilizing the laser to the 461-nm transition of strontium, using a heat pipe, and then forming a magneto-optic trap of strontium from a Zeeman-slowed atomic beam. PMID- 15119622 TI - Frequency locking of tunable diode lasers to a rubidium-stabilized ring-cavity resonator. AB - We demonstrate a technique for locking the frequency of a tunable diode laser to a ring-cavity resonator. The resonator is stabilized to a diode laser that is in turn locked to an atomic transition in rubidium, thus giving it absolute frequency calibration. The principal advantage of the ring-cavity design is that there is no feedback destabilization of the laser. The cavity has a free-spectral range of 1.3 GHz and Q of approximately 35, which provides robust locking of the laser. The locked laser is able to track large scans of the cavity. PMID- 15119623 TI - Calculation of the radiation trapping force for laser tweezers by use of generalized Lorenz-Mie theory. I. Localized model description of an on-axis tightly focused laser beam with spherical aberration. AB - Calculation of the radiation trapping force in laser tweezers by use of generalized Lorenz-Mie theory requires knowledge of the shape coefficients of the incident laser beam. The localized version of these coefficients has been developed and justified only for a moderately focused Gaussian beam polarized in the x direction and traveling in the positive z direction. Here the localized model is extended to a beam tightly focused and truncated by a high-numerical aperture lens, aberrated by its transmission through the wall of the sample cell, and incident upon a spherical particle whose center is on the beam axis. We also consider polarization of the beam in the y direction and propagation in the negative z direction to be able to describe circularly polarized beams and reflected beams. PMID- 15119624 TI - Calculation of the radiation trapping force for laser tweezers by use of generalized Lorenz-Mie theory. II. On-axis trapping force. AB - The efficiency of trapping an on-axis spherical particle by use of laser tweezers for a particle size from the Rayleigh limit to the ray optics limit is calculated from generalized Lorenz-Mie light-scattering theory and the localized version of a Gaussian beam that has been truncated and focused by a high-numerical-aperture lens and that possesses spherical aberration as a result of its transmission through the wall of the sample cell. The results are compared with both the experimental trapping efficiency and the theoretical efficiency obtained from use of the localized version of a freely propagating focused Gaussian beam. The predicted trapping efficiency is found to decrease as a function of the depth of the spherical particle in the sample cell owing to an increasing amount of spherical aberration. The decrease in efficiency is also compared with experiment. PMID- 15119625 TI - Passively Q-switched microlaser performance in the presence of pump-induced bleaching of the saturable absorber. AB - Unabsorbed pump light in passively Q-switched microlasers leads to suboptimal pulse generation by bleaching the saturable absorber. This mechanism increases the effective unsaturated transmission of the absorber, which leads to a change in the system dynamics that results in increased pulse durations and decreased pulse energies. We report experimental evidence of pump-induced bleaching of the saturable absorber, an increase in the pulse duration from 360 to 880 ps, and develop a simple analytical treatment that includes this effect within the framework of existing passive Q-switching models. PMID- 15119626 TI - Efficient mode transformations of degenerate Laguerre-Gaussian beams. AB - We present an approach for efficient conversion of a single-high-order-mode distribution from a laser to a nearly Gaussian distribution and vice versa. It is based on dividing the high-order mode distribution into equal parts that are then combined together coherently. We implement our approach with several optical arrangements that include a combination of discrete elements and some with single interferometric elements. These arrangements are analyzed and experimentally evaluated for converting the TEM01 mode distribution with Mx(2) = 3 to a nearly Gaussian beam with Mx(2) = 1.045 or Mx(2) = 1.15. The basic principle, design, and experimental results obtained with several conversion arrangements are presented. The results reveal that conversion efficiency is typically greater than 90%, compared with theoretical ones. In addition, some arrangement is exploited for converting the fundamental Gaussian-beam distribution into the TEM01 mode distribution. PMID- 15119627 TI - Penetration of high-intensity Er:YAG laser light emitted by IR hollow optical fibers with sealing caps in water. AB - The penetration depth in water was measured for Er:YAG laser light in a high density relevant to clinical applications. Various types of focusing elements were used to guide the light efficiently. We found that the transmission distance depended strongly on the beam shape in the water. When we used a plano-convex type of focusing cap, the penetration depth was larger than that when a dome- or ball-type cap were used. PMID- 15119628 TI - Effects of noise on lidar data inversion with the backward algorithm. AB - The lidar data-inversion algorithm widely known as the Klett method (and its more elaborate variants) has long been used to invert elastic-lidar data obtained from atmospheric sounding systems. The Klett backward algorithm has also been shown to be robust in the face of uncertainties concerning the boundary condition. Nevertheless electrical noise at the photoreceiver output unavoidably has an impact on the data-inversion process, and describing in an explicit way how it affects retrieval of the atmospheric optical coefficients can contribute to improvement in inversion quality. We examine formally the way noise disturbs backscatter-coefficient retrievals done with the Klett backward algorithm, derive a mathematical expression for the retrieved backscatter coefficient in the presence of noise affecting the signal, and assess the noise impact and suggest ways to limit it. PMID- 15119629 TI - Special visual effect of art glazes explained by the radiative transfer equation. AB - We present the first modeling of the light scattered by a paint layer in a bidirectional configuration. The studied medium is composed of small concentrated pigments embedded in an oil binder. The color is modulated by changing the number of paint layers, called glazes. The radiative transfer equation is established for incoherent light scattered by the pigments with use of a collimated illumination. The equation is solved by use of the auxiliary function method. This new method, applied here for the first time to a practical case, allows for exact computations of the scattered flux for any incident and collected directions. Spectroscopic and goniometric measurements are implemented in bidirectional and back-scattered configurations. The excellent agreement between the measurement and the simulation validates the assumptions used for the glaze model and proves the effectiveness of the auxiliary function method. PMID- 15119630 TI - Comparison of nanosecond and picosecond excitation for two-photon laser-induced fluorescence imaging of atomic oxygen in flames. AB - Two-photon laser-induced fluorescence (LIF) imaging of atomic oxygen is investigated in premixed hydrogen and methane flames with nanosecond and picosecond pulsed lasers at 226 nm. In the hydrogen flame, the interference from photolysis is negligible compared with the LIF signal from native atomic oxygen, and the major limitations on quantitative measurements are stimulated emission and photoionization. Excitation with a nanosecond laser is advantageous in the hydrogen flames, because it reduces the effects of stimulated emission and photoionization. In the methane flames, however, photolytic interference is the major complication for quantitative O-atom measurements. A comparison of methane and hydrogen flames indicates that vibrationally excited CO2 is the dominant precursor for laser-generated atomic oxygen. In the methane flames, picosecond excitation offers a significant advantage by dramatically reducing the photolytic interference. The prospects for improved O-atom imaging in hydrogen and hydrocarbon flames are presented. PMID- 15119631 TI - Will modulation of cell death increase PD technique survival? PMID- 15119632 TI - Effect of glucose degradation products on the peritoneal membrane in a chronic inflammatory infusion model of peritoneal dialysis in the rat. AB - BACKGROUND: Long-term use of the peritoneal membrane as a dialyzing membrane is hampered by its eventual deterioration. One of the contributing factors is glucose degradation products (GDPs) in the dialysis solution. In this study, we evaluated the effect of a low GDP solution on peritoneal permeability, the structural stability of the peritoneal membrane, and vascular endothelial growth factor (VEGF) production in a chronic inflammatory infusion model of peritoneal dialysis (PD) in the rat. METHODS: Male Sprague-Dawley rats were divided into 3 groups: a conventional solution group (group C, n = 12), a test solution group (group T, n = 12), and a normal control group (group NC, n = 8). Group T rats were infused with low GDP solution (2.3% glucose solution with two compartments), and group C rats with conventional dialysis solution (2.3% glucose solution), adjusted to pH 7.0 before each exchange. Animals were infused through a permanent catheter with 25 mL of dialysis solution. In both groups, peritoneal inflammation was induced by infusing dialysis solution supplemented with lipopolysaccharide on days 8, 9, and 10 after starting dialysate infusion. Peritoneal membrane function was assessed before and 6 weeks after initiating dialysis using the 1-hour peritoneal equilibration test (PET) employing 4.25% glucose solution. Both VEGF and transforming growth factor beta1 (TGFbeta1) in the dialysate effluent were measured by ELISA. The number of vessels in the omentum was counted after staining with anti-von Willebrand factor, and the thickness of submesothelial matrix of the trichrome-stained parietal peritoneum was measured. Peritoneal tissue was analyzed for VEGF protein using immunohistochemistry. RESULTS: At the end of 6 weeks, the rate of glucose transport (D/D0, where D is glucose concentration in the dialysate and D0 is glucose concentration in the dialysis solution before it is infused into the peritoneal cavity) was higher in group T (p < 0.05) than in group C. Dialysate-to-plasma ratio (D/P) of protein was lower in group T (p < 0.05) than in group C; D/P(urea), D/P(sodium), and drain volumes did not differ significantly between groups C and T. Dialysate VEGF and TGFbeta levels were lower in group T (p < 0.05) than in group C. Immunohistochemical studies also revealed less VEGF in the peritoneal membranes of group T. There were significantly more peritoneal blood vessels in group C (p < 0.05) than in group T, but the thickness of submesothelial matrix of the parietal peritoneum was not different between the two groups. The VEGF levels in the dialysate effluent correlated positively with the number of blood vessels per field (r = 0.622, p < 0.005). CONCLUSION: Using a chronic inflammatory infusion model of PD in the rat, we show that dialysis with GDP-containing PD fluid is associated with increased VEGF production and peritoneal vascularization. Use of low GDP solutions may therefore be beneficial in maintaining the function and structure of the peritoneal membrane during long-term PD. PMID- 15119633 TI - The effects of peritoneal dialysis solutions on peritoneal host defense. AB - Conventional peritoneal dialysis fluid (PDF) is a bioincompatible solution owing to the acidic pH, the high glucose concentrations and the associated hyperosmolarity, the high lactate concentrations, and the presence of glucose degradation products (GDPs). This unphysiologic composition adversely affects peritoneal host defense and may thus contribute to the development of PD-related peritonitis. The viability of polymorphonuclear leukocytes, monocytes, peritoneal macrophages, and mesothelial cells is severely depressed in the presence of conventional PDF. In addition, the production of inflammatory cytokines and chemoattractants by these cells is markedly affected by conventional PDF. Further, conventional PDF hampers the recruitment of circulating leukocytes in response to an infectious stimulus. Finally, phagocytosis, respiratory burst, and bacterial killing are markedly lower when polymorphonuclear leukocytes, monocytes, and peritoneal macrophages are exposed to conventional PDF. Although there are a few discrepant results, all major PDF components have been implicated as causative factors. Generally, novel PDF with alternative osmotic agents or with alternative buffers, neutral pH, and low GDP content have much milder inhibitory effects on peritoneal host defense. Clinical studies, however, still need to demonstrate their superiority with respect to the incidence of PD-related peritonitis. PMID- 15119634 TI - Transmigration of blood leukocytes into the peritoneal cavity is related to the upregulation of ICAM-1 (CD54) and Mac-1 (CD11b/CD18) adhesion molecules. AB - BACKGROUND: Migration of blood leukocytes into the peritoneal cavity of patients treated with peritoneal dialysis appears to be an important mechanism to prevent and fight peritonitis. To study the role of adhesion molecules in the process of leukocyte transmigration, we compared the expression of several adhesion receptors between peripheral blood monocytes and macrophages isolated from overnight dwell effluents. METHODS: The study was performed in 12, stable, infection-free patients treated with continuous ambulatory peritoneal dialysis (CAPD) and in 9 patients during peritonitis. In another set of experiments, we analyzed the expression of these molecules on blood leukocytes in 10 predialysis chronic renal failure (CRF) patients and 9 healthy controls. Peritoneal cells from an 8-hour dwell were isolated by centrifugation. Expression of adhesion receptors CD11a, CD11b, CD18, CD49d, and CD54 on blood and peritoneal leukocytes was measured using flow cytometry. RESULTS: In macrophages from the uninfected effluents, expression of both subunits of Mac-1 integrin receptor (CD11b and CD18) and intercellular adhesion molecule (ICAM)-1 receptor (CD54) was upregulated compared to peripheral blood monocytes from the same patients. The median value of mean fluorescence intensity in blood and effluent was 760.3 versus 1085.8 for CD11b (p = 0.013), 288.8 versus 448.6 for CD18 (p = 0.003), and 186.1 versus 365.7 for CD54 (p = 0.001). The same adhesion receptors were also significantly upregulated on peritoneal macrophages and neutrophils during peritonitis compared to blood leukocytes. Blood leukocytes from CAPD and CRF patients showed higher expression of CD54 and CD49d molecules compared to leukocytes from healthy controls. CONCLUSIONS: These data suggest that transmigration of blood leukocytes into the peritoneal cavity during uncomplicated dialysis and in peritonitis is related to selective upregulation of ICAM-1 (CD54) and Mac-1 (CD18/CD11b) receptors. PMID- 15119635 TI - Catabolism of newly synthesized decorin in vitro by human peritoneal mesothelial cells. AB - OBJECTIVE: Previous studies have shown that decorin and biglycan account for over 70% of the proteoglycans (PGs) synthesized by human peritoneal mesothelial cells (HPMCs). Since these PGs are involved in the control of cell growth, cell differentiation, and matrix assembly, we investigated their turnover in cultured HPMCs. METHODS: Confluent HPMCs were metabolically labeled with [35S]-sulfate and the labeled products isolated from the cell medium and the cell layer characterized by sensitivity to bacterial eliminases. Experiments were undertaken with exogenous labeled decorin, and its metabolic state was studied. RESULTS: In a 24-hour labeling period, 75% of the newly synthesized chondroitin sulfate/dermatan sulfate (CS/DS) PGs appeared in the culture medium, the majority of which (90%) was decorin. In the cell layer, protein-free glycosaminoglycan (GAG) chains accounted for 21% of the total CS/DS at 24 hours and exhibited constant specific activity at 12-16 hours. The latter material was turned over with a half-life of approximately 2.5 hours. Exogenous decorin underwent receptor mediated endocytosis and subsequent intracellular degradation. Uptake but not degradation could be inhibited by heparin. CONCLUSIONS: HPMCs are distinguished by a rapid turnover of decorin. A characteristic metabolic feature is the existence of a large intracellular pool of protein-free DS-GAGs. Understanding the control of decorin turnover in HPMCs might lead to delineation of its potential role in both the physiology and pathophysiology of the membrane in PD patients. PMID- 15119636 TI - Impact of fill volume on peritoneal clearances and cytokine appearance in peritoneal dialysis. AB - BACKGROUND: Current adequacy guidelines for peritoneal dialysis encourage the use of large fill volumes for the attainment of small solute clearance targets. These guidelines have influenced clinical practice in a significant way, and adoption of higher fill volumes has become common in North America. Several studies, however, have challenged the relevance of increasing small solute clearance; this practice may result in untoward consequences in patients. OBJECTIVE: The present study was designed to explore the relationship between dialysate volume and the clearance of different sized molecules, fluid dynamics, and appearance of peritoneal cytokines. METHODS: Thirteen adult prevalent patients on continuous ambulatory peritoneal dialysis were studied. Three different dialysate volumes (2.0, 2.5, and 3.0 L) were infused on consecutive days in a random order. Several measurements of peritoneal fluid dynamics (intraperitoneal pressure, net ultrafiltration, fluid absorption), solute clearances (urea, creatinine, beta2 microglobulin, albumin, IgG, and transferrin), and appearance of interleukin-6 and tumor necrosis factor alpha (TNFalpha) were assessed. RESULTS: Increase in dialysate fill volume (from 2 to 2.5 to 3 L) was examined in relationship to body surface area (BSA). The dialysate volume/BSA (DV/BSA) ratio increased from 1262 to 1566 to 1871 mL/m2 on 2.0, 2.5, and 3.0 L dialysate volumes, respectively. In parallel, diastolic blood pressure increased from 82.7 +/- 8.8 to 87.0 +/- 9.5 to 92 +/- 8.3 mmHg (p < 0.05). Net ultrafiltration rate also increased, from 0.46 +/ 0.48 to 0.72 +/- 0.42 to 0.97 +/- 0.49 mL/minute (p < 0.01), despite a concomitant increase in fluid absorption, from 1.05 +/- 0.34 to 1.21 +/- 0.40 to 1.56 +/- 0.22 mL/min (p < 0.01). Urea peritoneal clearance increased from 8.27 +/ 0.68 to 9.92 +/- 1.6 to 12.98 +/- 4.03 mL/min (p < 0.01); creatinine peritoneal clearance increased from 6.69 +/- 1.01 to 7.64 +/- 1.12 to 8.69 +/- 1.76 mL/min (p < 0.01). Clearance of the other measured molecules did not change. Appearance of interleukin-6 increased 17% and 43% (p < 0.01), and TNFalpha appearance increased 14% and 50% (p < 0.01) when dialysate volumes of 2.5 and 3.0 L were used, compared with 2.0 L. CONCLUSIONS: These results show that, with higher values of DV/BSA ratio, small solute peritoneal clearance is increased, but clearances of large molecules remain unchanged. With the use of higher volumes, fluid absorption rate and the appearance of proinflammatory cytokines in the dialysate are increased. PMID- 15119638 TI - Abnormalities of body composition in peritoneal dialysis patients. AB - OBJECTIVES: Body composition changes occur in peritoneal dialysis (PD) due to abnormalities in nutrition and hydration. We investigated abnormalities of nutrition and hydration in PD patients compared with healthy controls by measurement of total body potassium (TBK) and body water compartments. DESIGN: Cross-sectional comparison study. METHODS: We measured TBK--an indicator of body cell mass--by whole body counting, total body water (TBW) by deuterium oxide dilution, and extracellular water (ECW) by bromide dilution in 29 PD patients and 32 controls. RESULTS: The absolute mean value of TBK for PD patients was not significantly lower than in controls. The ratios of observed TBK to predicted TBK from prediction formulas were compared. Equations used were those of Boddy, Bruce, Burkinshaw, and Ellis and our own equation derived from a local control database (Leeds). Observed/predicted ratios of TBK were significantly less in PD than in control subjects for all equations. Water volumes did not differ between PD and control groups. Observed/predicted ratios for TBK in PD patients correlated with serum potassium (Boddy r = 0.355, p = 0.06; Bruce r = 0.411, p < 0.05; Burkinshaw r = 0.457, p < 0.01; Leeds r = 0.412, p < or = 0.05; Ellis r = 0.356, p = 0.06) and tended to correlate with serum albumin (Bruce r = 0.343, p = 0.07; Burkinshaw r = 0.421, p < 0.05; Leeds r = 0.357, p = 0.06; Ellis r = 0.310, p = NS). There was no relationship with serum potassium in controls. Serum albumin in PD correlated with TBK (r = 0.445, p < 0.02), TBK/height (r = 0.419, p < 0.05), TBK/weight (r = 0.554, p = 0.002), and TBK/TBW (r = 0.586, p = 0.0001). Extracellular water/intracellular water (ECW/ICW) was inversely related to TBK (r = -0.455, p < 0.02 in PD; r = -0.387, p < 0.05 in controls) and to TBK/height (r = -0.446, p < 0.02 in PD; r = -0.411, p = 0.02 in controls). TBK/weight reduced with age in PD (r = -0.445, p < 0.02), as did TBK/TBW in PD (r = -0.463, p < 0.02). ECW/ICW tended to increase with age in PD (r = 0.351, p = 0.06). CONCLUSIONS: Observed/predicted ratio of TBK is reduced in PD patients relative to healthy controls, indicating reduced body cell mass. Serum albumin and potassium reflect TBK indices in PD. Body water volumes did not differ between PD and controls, implying no overall abnormality in hydration in the PD group. However, ECW is relatively increased compared to ICW with decreasing TBK indices, suggesting relative ECW expansion with reduction in body cell mass. PMID- 15119637 TI - Bone mineral density: serum markers of bone turnover and their relationships in peritoneal dialysis. AB - BACKGROUND: The usefulness of bone mass measurements and bone turnover markers to estimate the risk of fracture and the type of underlying renal osteodystrophy are not well established in patients on peritoneal dialysis (PD). OBJECTIVE: To assess bone mass using total and regional bone densitometry in a group of patients on PD and to determine if serum markers of bone turnover identify patients with low bone mass. METHODS: Bone densitometry was studied by dual energy x-ray absorptiometry (DEXA), and bone turnover using several serum markers, in 65 patients on PD. Bone mass was classified as normal, osteopenic, or osteoporotic according to World Health Organization criteria based on bone mineral density (BMD) T scores. RESULTS: T scores in the osteopenia range were present at the lumbar spine (LS) in 44.6% (45% of men and 44.4% of women) of patients and at the femoral neck (FN) in 56.9% (55% of men and 58% of women). T scores in the osteoporosis range were present at the LS in 13.8% of patients (10% of men and 15.5% of women) and at the FN in 21.5% (30% of men and 17.7% of women). Patients with BMD T scores in the osteoporosis range at both regions had increased serum intact parathyroid hormone (iPTH) levels compared to patients in the osteopenic/normal range. Bone mineral content in the whole skeleton (TBMC) correlated negatively with iPTH (r = -0.34) and with total time on dialysis (r = 0.26); in multivariate analysis, only iPTH correlated negatively with TBMC (B = 0.26, p = 0.03). No correlations were found between the other bone markers and BMD T scores at the FN or LS. There were no significant differences in absolute BMD or BMD T scores at the LS or FN between patients with and patients without fractures. CONCLUSIONS: BMD T scores in the osteopenia/osteoporosis range were observed at the LS in 58.4% of these patients on PD and at the FN in 78.4%. TBMC correlated negatively with iPTH. There were no correlations between markers of bone turnover and bone mass measurements at the two skeletal regions, although patients with BMD T scores in the osteoporosis range had increased serum iPTH levels. Bone mass measurements were not different between patients with and patients without fractures. PMID- 15119639 TI - The Initiating Dialysis Early and Late (IDEAL) study: study rationale and design. AB - OBJECTIVES: The primary objective of the IDEAL study is to determine whether the timing of dialysis initiation has an effect on survival in subjects with end stage renal disease (ESRD). The secondary objectives are to determine the impact of "early start" versus "late start" dialysis on nutritional and cardiac morbidity, quality of life, and economic cost. DESIGN: Prospective multicenter randomized controlled trial. Patients are randomized to commence dialysis at a glomerular filtration rate (by Cockcroft-Gault) of either 10-14 mL/minute/1.73 m2 ("early start") or 5-7 mL/min/1.73 m2 ("late start"), with stratification for dialysis modality (hemodialysis vs peritoneal dialysis), study center, and the presence or not of diabetes mellitus. SETTING: Dialysis units throughout Australia and New Zealand. PATIENTS: Patients with ESRD commencing chronic dialysis therapy. OUTCOME MEASURES: Three years from randomization, all-cause mortality, morbidity, and economic impact; structural and functional cardiac status, nutritional state, and quality of life will be assessed. RESULTS: To date, 388 patients of a minimum 800 patients have been entered and randomized into the study. Current recruitment rates suggest sufficient patients will be enrolled by December 2004 and follow-up completed by December 2007. CONCLUSIONS: The IDEAL study will provide evidence for the optimal time to commence dialysis. PMID- 15119640 TI - Methods to determine drain volume for peritoneal dialysis clearances. AB - OBJECTIVE: To compare the accuracy and convenience of 3 methods for measuring drain volume for peritoneal dialysis (PD) clearance studies. DESIGN: Prospective comparison of both automated PD (APD) and continuous ambulatory PD (CAPD) clearance study methods. SETTING: Adults > or = 18 years old at 2 dialysis clinics. PATIENTS: 28 PD patients with 43 clearance studies, 15 on CAPD and 28 on APD. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Drain volume was determined by 3 methods for each study: (1) graduate-measured volume using a 2-L graduated cylinder; (2) weighed volume, with and without bag weight, using a digital floor scale or spring scale; (3) cycler-measured volume using the initial drain and ultrafiltration indicated by the cycler, plus the prescribed inflow volume without the last fill. RESULTS: There was no statistically significant difference in volumes using the 3 methods studied (all p > 0.89 for APD, all p > 0.97 for CAPD). Effluent volume was more accurate with the weight of the bag subtracted. CONCLUSION: The most convenient and a precise method for APD is to determine the effluent drain volume using the prescription and total ultrafiltration and initial drain, as measured by the cycler. For CAPD, using the weight of drained bags is accurate but the weight of the empty bag must be subtracted. These approaches have the least risk of exposing staff to body fluids. PMID- 15119641 TI - Fibrin glue is useful in preventing early dialysate leakage in children on chronic peritoneal dialysis. AB - OBJECTIVE: To assess if application of fibrin glue sealant to the peritoneal cuff suture is useful in the prevention of early dialysate leakage in children with end-stage renal disease on chronic peritoneal dialysis (CPD). DESIGN: Single center, open-label, prospective randomized study. SETTING: University Pediatric Hospital. METHODS: 52 catheters were implanted in 45 children (mean age 6.2 +/- 4.5 years). Catheters were randomly assigned to either the control group or the sealant group. In the latter group, 1 mL of fibrin glue sealant was applied to the peritoneal cuff suture. 18 catheters were used for the first time within 5 days after implantation (early-used catheters). Leakage, exit-site or tunnel infection, peritonitis, and adverse secondary effects were evaluated during the initial 60 days after implantation. RESULTS: No adverse secondary effects were seen after the application of the fibrin glue sealant. The incidence of exit/tunnel infection and peritonitis was similar in the two groups. The incidence of leakage was significantly lower in the sealant group (p < 0.02). In the early-used catheters, leakage was detected in 9% of the catheters in the sealant group and in 57% of the control group (p < 0.05). CONCLUSIONS: The application of 1 mL of fibrin glue to the peritoneal cuff suture prevented early dialysate leakage without secondary adverse effects in children on CPD. PMID- 15119643 TI - An unusual neurological complication in a CAPD patient. PMID- 15119642 TI - Significance of slime as virulence factor in coagulase-negative staphylococcus peritonitis in CAPD. PMID- 15119644 TI - Peritoneal dialysis-associated peritonitis caused by Staphylococcus auricularis. PMID- 15119645 TI - Klebsiella oxytoca and polymicrobial infection in peritoneal dialysis-related peritonitis. PMID- 15119646 TI - The icodextrin black line sign to confirm a pleural leak in a patient on peritoneal dialysis. PMID- 15119647 TI - Early morning blues--a complication of icodextrin. PMID- 15119648 TI - No beneficial effect of icodextrin on blood glucose control. PMID- 15119649 TI - Literature: March-April 2004. PMID- 15119650 TI - Operative endoscopy will replace almost all open procedures. PMID- 15119651 TI - 1000 office-based hysteroscopies prior to in vitro fertilization: feasibility and findings. AB - BACKGROUND AND OBJECTIVES: Hysteroscopy offers diagnostic accuracy and the ability to treat uterine pathology, but practitioners may be reluctant to perform it without a high index of suspicion because it traditionally requires an operating room. This study reviews the findings and feasibility of office-based diagnostic and operative hysteroscopy in an unselected in vitro fertilization (IVF) population to evaluate whether first-line hysteroscopy should be recommended. METHODS: One thousand consecutive infertile patients scheduled for in vitro fertilization underwent office hysteroscopy. A rigid 20-degree 5-mm hysteroscope, with an operative channel for grasping forceps, scissors, or coaxial bipolar electrode was used. Operative findings, complications, and patient tolerance were noted. RESULTS: Sixty-two percent of patients had a normal uterine cavity. Thirty-two percent had endometrial polyps. Other pathology included submucous fibroids (3%), intrauterine adhesions (3%), polypoid endometrium (0.9%), septum (0.5%) retained products of conception (0.3%), and bicornuate uterus (0.3%). The pathology was treated in all patients without complication. CONCLUSIONS: When hysteroscopy is routinely performed prior to in vitro fertilization, a significant percentage of patients have uterine pathology that may impair the success of fertility treatment. Patient tolerance, safety, and the feasibility of simultaneous operative correction make office hysteroscopy an ideal procedure. PMID- 15119652 TI - Mayo Clinic Scottsdale Experience with laparoscopic nephroureterectomy. AB - OBJECTIVES: To evaluate the efficacy of laparoscopic nephroureterectomy for patients with transitional cell carcinoma of the upper urinary tract. METHODS: Eighteen patients underwent attempted transperitoneal laparoscopic nephroureterectomy between June 2000 and October 2002. Mean patient age was 67.5 years. The specimen was removed intact through a 7- to 9-cm extraction incision in the lower midline. In the majority of patients, the distal ureter was dissected through the extraction incision. RESULTS: Sixteen cases were completed laparoscopically. Two cases required conversion to an open procedure. In these cases, dense fibrosis was present around the renal hilum preventing further dissection. The mean operative time was 180 minutes, and the mean estimated blood loss was 160 mL. The mean length of stay was 3.3 days. Complications included the 2 conversions, and 1 patient with a postoperative Mallory Weiss tear. No port site or distant metastasis occurred; however, 1 patient developed a retroperitoneal recurrence. CONCLUSION: Laparoscopic nephroureterectomy is an alternative to open nephroureterectomy. Cases with high-stage and grade may cause the laparoscopic dissection to be difficult. The extraction incision allows for easy dissection of the distal ureter. PMID- 15119653 TI - Laparoscopy in the management of pediatric intraabdominal tumors. AB - BACKGROUND AND OBJECTIVES: Unresectable intraabdominal tumors pose a challenge to the pediatric oncology team. Tumor tissue is needed for diagnostic and prognostic analyses. Laparotomy is quite invasive and computerized tomography-guided needle biopsies, though less invasive, may not procure enough tissue for a complete analysis. This study aimed to evaluate our experience with laparoscopy in managing these tumors. METHODS: Nine children underwent laparoscopy: 6 for primary tumor biopsy, 2 for tumor resection, and 1 for tumor dissection followed by laparotomy for tumor resection. RESULTS: The study included 7 females and 2 males with a median age of 5 years (range, 3 to 15 years). Three patients had neuroblastoma, 2 had mature ovarian teratomas, 2 had rhabdomyosarcoma, 1 had erythrophagocytic histiocytosis, and 1 had T-cell hepatosplenic lymphoma. In each case, adequate tissue was procured for diagnostic and prognostic analyses. No perioperative complications occurred. Both patients with ovarian teratomas are well. Four patients continue to receive anti-cancer therapy. The patient with erythrophagocytic histiocytosis underwent a cord-blood stem cell transplant. One patient with neuroblastoma was lost to follow-up, and 1 patient with rhabdomyosarcoma died of progressive disease. CONCLUSIONS: Laparoscopy is a valuable technique in the management of pediatric intraabdominal tumors. It allows for tumor biopsy under direct vision, and adequate tissue is procured for all analyses. Moreover, it allows the surgeon to dissect the tumor and determine resectability. PMID- 15119654 TI - Efficacy of laparoscopic cholecystectomy in acalculous gallbladder disease: long term follow-up. AB - OBJECTIVE: Our aim was to determine the efficacy of laparoscopic cholecystectomy in symptomatic patients with ultrasound negative and abnormal gallbladder ejection fractions; Patients with gallbladder ejection fractions less than 35% on hepatobiliary scan were offered laparoscopic cholecystectomy. METHODS: Between January 1995 and January 2001, 1564 patients underwent laparoscopic cholecystectomy at our institution: 256 were confirmed to have acalculous gallbladder disease by pathology report and reconfirmation of abnormal hepatobiliary scan data. A 30-day postoperative follow-up was obtained by retrospective medical record review. For this study, we contacted all 256 patients by mail questionnaire and followed up on nonresponders with telephone interviews; we also reviewed hospital records to verify preoperative symptom patterns. The survey was completed by 154 patients (60%): 48 (31%) by mail and 106 (69%) by telephone interviews. The study included 115 (75%) female and 39 (25%) male patients, and the average age was 42 years (range, 13 to 95). All hepatobiliary laboratory parameters were normal pre- and postoperatively. The survey was completed in December 2001, 1 to 5 years postoperatively (mean 3 years). RESULTS: Preoperatively, 142 patients (92%) had right upper quadrant pain, 114 (74%) had nausea, 88 (57%) had vomiting, 120 (73%) had heartburn, and 118 (77%) had food intolerance. In a 30-day postoperative period, these numbers had reduced to 48 (37%), 14 (90%), 8 (5%), 22 (14%), and 34 (22%), respectively. had laparoscopic cholecystectomy, and 95% stated that they would recommend laparoscopic cholecystectomy to other patients. CONCLUSION: This study shows that patients with acalculous gallbladder disease benefit from laparoscopic cholecystectomy. PMID- 15119655 TI - An evaluation of transanal endoscopic microsurgery for rectal adenoma and carcinoma. AB - BACKGROUND: Transanal endoscopic microsurgery was introduced in the early 1980s. Since then, increasing numbers of rectal adenomas are being excised by this technique. The aim of this study was to evaluate our institution's experience with transanal endoscopic microsurgery for rectal adenoma and carcinoma. METHODS: Seventy-five patients (adenomas, n = 58) underwent more than 90 TEM resections over a period of 5 years. RESULTS: Postoperative complications were minimal with 3% (n = 2) in the adenoma group requiring transfusion and 0% 30-day mortality. One patient in each group developed transient fecal incontinence. During the follow-up period, 6 patients (10%) in the adenoma group underwent further local resections for their recurrences. Two patients in the carcinoma group (1 each of pathological T1 and T2 stage) developed recurrence at 24 months. A female with a T2 tumor was found to have an inoperable lesion and underwent sigmoid colostomy. Five of 17 patients had postoperative radiotherapy, and 2 patients developed radiation enteritis. Four patients died during follow-up due to unrelated reasons. CONCLUSION: The transanal endoscopic microsurgery technique appears to be safe and associated with minimal morbidity. Careful selection of patients with thorough preoperative assessment is necessary for carcinoma patients. Patients with T1 lesions and favorable histology should only be considered for curative resection by this technique. PMID- 15119656 TI - Adhesion formation after laparoscopic ventral incisional hernia repair with polypropylene mesh: a study using abdominal ultrasound. AB - OBJECTIVE: Laparoscopic repair of ventral incisional hernias is feasible and safe. Polypropylene mesh is often preferred because of its ease of handling and lower cost. Complications like adhesion and fistula formation can occur. The goal of this study was to determine whether bowel adhesions and their attendant complications could be prevented by interposition of omentum. METHODS: Thirty patients underwent laparoscopic ventral incisional hernias repair with polypropylene mesh. Omentum was always positioned over the loops of bowel for protection. At a mean follow-up of 14 months, 20 patients underwent ultrasonic examination using the previously described visceral slide technique to detect adhesions. RESULTS: The mean size of the hernias in the study was 50.3 cm2, and the mean size of the mesh applied was 275 cm2. Thirteen patents (65%) had no sonographically detectable adhesions. Five patients demonstrated adhesions between the mesh and omentum, 1 patient developed adhesions between the left lobe of the liver and the mesh, and only 1 case of bowel adhesion to the edge of the mesh was found. CONCLUSION: Laparoscopic ventral incisional hernias repair with polypropylene mesh and omental interposition is not associated with visceral adhesions in the majority of patients. Polypropylene mesh can be used safely when adequate omental coverage is available. PMID- 15119657 TI - The incidence of port-site metastases in gynecologic cancers. AB - OBJECTIVES: To determine the incidence of port-site metastases in patients undergoing laparoscopic procedures for gynecologic cancers. METHODS: The charts of patients treated by laparoscopy for diagnosis, treatment, or staging of gynecologic cancers by the academic faculty attending physicians were studied from July 1, 1997 to June 30, 2001. No patient without a histological or cytological diagnosis of cancer from the index procedure were included. Fisher's exact test was used for statistical analysis. RESULTS: Eighty-three patients were identified accounting for 87 procedures. Types of cancer treated included endometrial (39), ovarian (29), and cervical (14). Twenty procedures were performed for recurrence of ovarian or peritoneal cancer, and ascites was present in 10 cases. Port-site metastases occurred in 2 patients accounting for 8 sites. Five sites were diagnosed in a single patient 13 days after a second-look laparoscopy for stage IIIB ovarian cancer, and 3 sites were diagnosed in a patient 46 days after an interval laparoscopy for stage IIIC primary peritoneal cancer. Ascites was present in both patients. The overall incidences of port-site metastases per procedure and per port placed were 2.3% (2/87) and 2.4% (8/330), respectively. In patients with a recurrence of ovarian or peritoneal cancer, no port-site metastases (0/16) occurred in the absence of ascites, whereas 50% (2/4) of patients with ascites developed port-site metastases (P < .035). CONCLUSIONS: The overall incidence of port-site metastases in gynecologic cancers in our study was 2.3%. The risk of port-site metastases is highest (5%) in patients with recurrence of ovarian or primary peritoneal malignancies undergoing procedures in the presence of ascites. PMID- 15119658 TI - Fifty-five consecutive laparoscopic appendectomy procedures without conversion. AB - BACKGROUND: In patients with suspected appendicitis, laparoscopic appendectomy is gaining increasing acceptance primarily because it is associated with less postoperative pain and a shorter hospital stay. Experience with 55 consecutive laparoscopic appendectomies, performed without conversion by the same surgeon, is herein examined and analyzed. METHODS: The medical records of 55 consecutive patients with suspected appendicitis who underwent laparoscopic exploration (from 2000 to 2002) were analyzed for demographic information, clinical findings, laboratory/computed tomography scan results, intraoperative diagnosis, clinicopathologic correlation, complications, incidental findings, and operative time. RESULTS: Twenty-six males (47%) and 24 females (53%) underwent surgery. Mean age was 25.2 years (range, 6 years to 67 years). Computerized tomography scans obtained in 37 cases (74%) had a sensitivity of 86.7% and a specificity of 62.5%. Average length of stay was 2.3 days (median, 1 day). Average operating room time was 69 minutes (range, 40 to 173 minutes). Five patients experienced postoperative complications: 2 had intraperitoneal abscesses, 1 had urinary retention, and 2 had postoperative ileus. No operative conversions or postoperative wound infections occurred. CONCLUSION: Laparoscopy confirmed the clinical diagnosis of acute appendicitis and allowed the safe, effective treatment of both complicated and uncomplicated appendicitis with minimal hospitalization, recovery and convalescent times, and zero open conversion and wound infections. Laparoscopic appendectomy is advocated as the procedure of choice for patients with clinically suspected appendicitis. PMID- 15119659 TI - Perforated appendicitis: is laparoscopy safe? AB - OBJECTIVE: The purpose of this study was to demonstrate the safety and efficacy of laparoscopy in children with perforated appendicitis. METHODS: This is a retrospective review of consecutive patients under the age of 18, operated on for perforated appendicitis between September 1997 and December 1999. RESULTS: Sixty nine patients were operated on for perforated appendicitis. Eleven appendectomies were performed laparoscopically. Fifty-four patients underwent an open appendectomy. Four laparoscopic appendectomies were converted to an open procedure. The mean operative time was 79 minutes for the laparoscopic group, and 87 minutes for the open group. The mean length of hospital stay was 5.4 days versus 7.6 days for the laparoscopic and open groups, respectively. Neither of these differences was statistically significant. CONCLUSIONS: The above data support the use of laparoscopy in the management of perforated appendicitis in children. In conclusion, laparoscopy is as safe as open appendectomy. Laparoscopy is an effective alternative with a shorter length of hospital stay compared with that for an open appendectomy for perforated appendicitis in children. PMID- 15119660 TI - Laparoscopic appendectomy in children can be done as a fast-track or same-day surgery. AB - BACKGROUND: Laparoscopic surgery has reduced the length of hospital stay for common operations like cholecystectomy, gastric fundoplication, and appendectomy. We have noticed a reduction in length of hospital stay for children undergoing laparoscopic appendectomy. We, therefore, looked at our data to assess whether laparoscopic appendectomy in children could be performed as fast-track or same day surgery (< or = 24-hour postoperative stay). METHODS: We performed a retrospective review of the records of all children who underwent laparoscopic appendectomy for suspected appendicitis during a 3-year period (7/97 to 7/00). RESULTS: Laparoscopic appendectomy was performed in 79 children (44 boys and 35 girls), between 2 to 17 years of age (mean, 11 years). In 4 (5%) children with perforated appendicitis, the laparoscopic appendectomy was converted to an open appendectomy. At operation, 51 (64.5%) had acute appendicitis, 22 (27.8%) had perforated appendicitis, 4 (5%) had ruptured ovarian cysts, and 2 (2.5%) had no pathology. The median operative time was 54 minutes. Total length of stay for all 79 patients was a median of 58 hours, and median postoperative LOS was 35 hours. Complications included wound infection (2), abdominal abscess (4), drug rash (2), and epididymo-orchitis (1). In 57 (72%) children without perforated appendicitis, the total length of hospital stay was a median of 42 hours, while median postoperative length of stay was only 28 hours. Thirty-two (56%) children went home in < or = 24 hours following laparoscopic appendectomy. No significant morbidity was noted in the nonperforated group (drug rash, 1 fever > 24 hrs, 3); and no readmissions or reoperations were necessary on follow-up. CONCLUSION: Laparoscopic appendectomy is safe and effective for treating children with appendicitis. Laparoscopic appendectomy may be safely performed as fast-track or same-day surgery, in select children without perforated appendicitis, with a postoperative stay of < or = 24 hours. PMID- 15119661 TI - Safety of cystic duct clipping in healthy and cirrhotic livers: a cadaveric study. AB - BACKGROUND AND OBJECTIVES: Biliary leakage through the cystic duct stump due to clip dislodgement has been a concern since the advent of the laparoscopic cholecystectomy. The authors proposed a cadaveric model to test the safety of cystic duct clipping in a hypertensive biliary tract in healthy and cirrhotic livers. METHODS: Twenty fresh cadavers were studied (5 cirrhotic, 15 healthy). Open cholecystectomy was performed and the cystic duct clipped with commercially available titanium clips. The distal common bile duct was catheterized to allow infusion of water and pressure measurement. RESULTS: Increased pressure in the bile duct resulted in back diffusion into the liver, preventing reaching high pressure levels. Only 1 clip was dislodged in this situation, in a cirrhotic liver with a large cystic duct. As a second experiment, the hepatic hilum was clamped to allow higher pressures of the biliary tree (500 mm Hg). In this situation, no clip was dislodged. CONCLUSIONS: We have established the safety of cystic duct clipping in healthy and cirrhotic livers; however, bigger clips or alternative methods to seal the duct may be necessary in larger ducts. PMID- 15119662 TI - Ischemic necrosis of small bowel following laparoscopic surgery. AB - BACKGROUND AND OBJECTIVE: Small bowel ischemia following laparoscopy was described recently as a rare fatal complication of the CO2 pneumoperitoneum. Of the 8 cases reported in the surgical literature, 7 were fatal, 1 was not. In this report, we describe the first gynecological case. METHODS: A 34-year-old woman who underwent laparoscopy with extensive adhesiolysis and myolysis was re admitted with an acute abdomen on postoperative day 4. Immediate laparotomy revealed acute peritonitis, extensive adhesions, and a 3-cm defect in the small bowel. Tissue examination showed ischemic necrosis of edematous, but essentially normal, bowel mucosa. The postoperative course was extremely complicated. She was discharged after a 2-month hospital stay in the intensive care unit for rehabilitation. RESULTS: Data are available on 7 patients (including ours). All procedures were described as uneventful. The intraabdominal pressure was set at 15 mm Hg when specified. Some abdominal pain occurred in all, nausea and vomiting in 4, diarrhea in 2, abdominal distention in 1, fever in none. Quick reintervention laparotomy was performed in 2 and delayed in 5 (up to 4 days). DISCUSSION: The CO2 pneumoperitoneum is a predisposing factor for intestinal ischemia as it reduces cardiac output and splanchnic blood flow. However, critical ischemia relies on underlying vasculopathy or an inciting event. CONCLUSION: Patient selection, maintaining intraabdominal pressure at 15 mm Hg or less, and intermittent decompression of the gas represent the best options for preventing this complication. PMID- 15119663 TI - Acute mesenteric venous thrombosis following laparoscopic Roux-en-Y gastric bypass. AB - BACKGROUND AND OBJECTIVES: Acute mesenteric venous thrombosis has not been previously reported as a complication following Roux-en-Y gastric bypass. METHODS: The authors present 3 cases from a single-center experience of over 1500 patients as well as a review of the literature. RESULTS: The presenting symptoms are nonspecific, and the diagnosis is often made after infarction of the intestine has occurred. A high index of clinical suspicion is required for timely diagnosis and treatment. A computed tomography scan combined with diagnostic laparoscopy are the gold standard diagnostic tests, and early anticoagulation is the optimal treatment. Diagnostic laparoscopy is essential to evaluate the degree of bowel ischemia and the need for resection. CONCLUSION: Acute mesenteric venous thrombosis following Roux-en-Y gastric bypass is a severe and potentially life threatening complication that requires early exploration and anticoagulation. PMID- 15119664 TI - Appendiceal enterobius vermicularis infestation associated with right-sided chronic pelvic pain. AB - Parasitic infestation is an uncommon cause of chronic pelvic pain among women of reproductive age. A case of chronic right-sided pelvic pain associated with appendiceal Enterobius vermicularis infestation was managed with appendectomy and antiparasitic therapy resulting in a complete resolution of symptoms. PMID- 15119665 TI - Laparoscopic appendectomy in a female patient with situs inversus: case report and literature review. AB - BACKGROUND: Situs inversus is an uncommon condition caused by a single autosomal recessive gene of incomplete penetration. A potential diagnostic dilemma can occur in the young female patient with a history of situs inversus who presents with pelvic pain. METHODS: A 32-year-old multiparous patient with a known history of situs inversus presented with complaints of pelvic pain. A medical history and full physical examination were indicative of possible endometriosis. RESULTS: The patient underwent an operative laparoscopy, which revealed stage II pelvic endometriosis based on the American Fertility Society Revised Classification for Endometriosis (R-AFS), with appendicular and periappendicular adhesions involving the cecum. Ablation of endometriosis and an appendectomy were performed. CONCLUSION: The authors believe the laparoscopic approach to an appendectomy is ideal in a patient with situs inversus and should be performed at the time of laparoscopy performed for another reason. PMID- 15119666 TI - Thoracoscopic excision of a paraesophageal bronchogenic cyst in a child. AB - BACKGROUND: Bronchogenic cysts are not uncommon in either children or adults. In children, they comprise approximately 6% of all mediastinal masses. Their presentation can range from an asymptomatic incidental finding to sudden respiratory distress. CASE REPORT: Video-assisted thoracoscopy was utilized to remove a bronchogenic cyst that was densely adherent to the adjacent esophagus in a child. This was accomplished with a Harmonic scalpel. The chest tube was removed on postoperative day 1, and the patient was discharged on postoperative day 2. An esophagogram obtained 2 weeks after surgery was normal, and the patient's preoperative symptoms had not returned. CONCLUSIONS: Bronchogenic cysts should be considered in the differential diagnoses for mediastinal masses at any age. Given their benign nature, thoracoscopy offers an excellent alternative to open thoracotomy for their removal. PMID- 15119667 TI - Expert videotape analysis and critiquing benefit laparoscopic skills training of urologists. AB - INTRODUCTION: Teaching laparoscopic skills has become the focus of the latest generation of hands-on laparoscopic courses. METHODS: Thirty-four practicing urologists, ages 31 to 61 years (mean, 46.6 years) with laparoscopic experience (range, 0 to 200, mean, 27.6 cases), 32 of whom had taken prior American Urological Association (AUA) laparoscopy courses, participated in an AUA sponsored hands-on laparoscopic skills course over a 2-day period in August 2002 or March 2003. They all took a knowledge assessment examination and performed standardized tasks (rope passing, ring placement, and laparoscopic suturing and knot tying) at the beginning and the end of the course with a videotape analysis and critique. Prior to the repeat-skills assessment, each participant was individually critiqued and instructed based on a videotape review of their initial performance. The urologists also participated in a porcine laboratory and a pelvic trainer session totaling 6 hours between skills assessments. None of the participants had performed significant laparoscopic suturing prior to the course. RESULTS: Using Wilcoxon's signed rank test, the participants improved from a mean of 119.32 seconds to 98.36 seconds with the rope pass (P = 0.0001), and with the ring placement from a mean of 9.70/minute to 12.09/minute (P = 0.0001). All participants had significantly fewer false passes (mean, 9.35 compared with 5.21) during repeat skills assessments (P = 0.0001). Participants improved from 0.54 sutures/minute to 1.22 sutures/ minute following the video critique and practice (P = 0.0001). Degree of laparoscopic experience (number of cases), age of the urologist, and precourse knowledge (examination score) had no significant bearing on results in the initial skills assessment or in the improvement of task time (Spearman correlation coefficients). CONCLUSION: Urologists with some laparoscopic experience (mean 27.6 cases) can improve laparoscopic skills using mentored videotape analysis and experience gained from a 2-day hands-on course. Prior knowledge, degree of experience, and urologist age had no significant bearing on performance in this setting. PMID- 15119668 TI - Laparoscopic suturing and knot tying: a comparison of standard techniques to a mechanical assist device. AB - BACKGROUND: Suturing and knot tying are basic skills for surgeons. Performing these tasks laparoscopically can be a tedious, time-consuming endeavor associated with much frustration. We evaluated a mechanically assisted suture and pretied knot device (Quik-Stitch) for performing the basic tasks of suturing and knot tying. METHODS: We performed a time study using 1) intracorporeal suturing and knot tying, 2) intracorporeal suturing and extracorporeal knot tying, and 3) a mechanically assisted suture and pretied knot device (Quik-Stitch). From September 2000 through March 2001, time trials were conducted using each of the different techniques. Three attending surgeons, one with much experience and 2 with less experience, and 2 chief residents, with the least experience, participated in the study. RESULTS: For the experienced surgeon, the average times for intracorporeal knot tying, extracorporeal knot tying, and knot tying with Quik-Stitch were 97.3, 103.9, and 67.7 seconds, respectively. For the less experienced surgeons, the times were 237.2, 224.3, and 92.5 seconds, respectively. For the least experienced group, the times were 265.3, 263.0, and 128.7 seconds, respectively. CONCLUSIONS: The mechanically assisted suture device and pretied knot (Quik-Stitch by PARE Surgical, Inc, Englewood, CO, USA) provides significant time-saving to surgeons regardless of experience and thus reduces operating room costs. Less experienced surgeons and surgeons in training benefited the most by the use of this device. PMID- 15119669 TI - Evaluation of specialized laparoscopic suturing and tying devices. AB - BACKGROUND: Laparoscopic suturing and tying constitute advanced minimally invasive surgery skills. Developing proficiency in the standard methods with needle drivers is often an arduous process. Recent advances in laparoscopic instrumentations has allowed for easier methods of suturing and tying. This study investigated the hypothesis that the use of a specialized suturing device and a specialized tying device allows inexperienced medical students to suture and tie laparoscopically. METHODS: Preclinical medical students who had not received any training in open or laparoscopic surgery were included in this investigation. Each student was given a 5-minute demonstration of a specialized suturing device and a specialized tying device. The medical students were not allowed to deploy either device before actual use. After the demonstration, each student was given the device to use in a porcine model. Times were recorded and a subjective grade was given for each student. RESULTS: Twenty medical students were involved in this study. All medical students were able to complete the task of suturing and tying. The average time to suture was 104.6 seconds and the average time to tying was 31.2 seconds. The average subjective performance grade was 90 (out of 100). CONCLUSION: Specialized devices are easy to learn and use for laparoscopic suturing and tying with minimal instruction even for inexperienced medical students. Even surgeons who are not well versed in laparoscopic surgery should be able to suture and tie with certain laparoscopic instruments. PMID- 15119670 TI - Training and assessment of laparoscopic skills. AB - Laparoscopic surgery is gaining popularity among the surgical community. While its prevalence expands, the need for reliable training and assessment tools is becoming increasingly important. Laparoscopic skills are not an innate behavior, nor can they be easily mimicked, and can only be acquired through hands-on training. A consensus exists among physicians that establishment and evaluation of technical skill in surgical training programs are inadequate and in need of improvement. A validated, reliable bench model that could train and assess could be standardized and provide numerous benefits including determination of which medical students should consider a career in surgery, valuable feedback to residents, a tracking mechanism of resident performance, a possible certification and recertification tool, and to allow for interinstitutional comparison. To this end, several potentially successful bench models testing dexterity, hand-eye coordination, and depth perception have been developed. A few models have been proven to be both valid and reliable indicators of technical skill. Although the future remains uncertain, enough groundwork has been laid to begin incorporating technical skill training and assessment into surgical training programs. PMID- 15119671 TI - Replacement of expensive, disposable instruments with old-fashioned surgical techniques for improved cost-effectiveness in laparoscopic hysterectomy. AB - OBJECTIVE: Patients demand that health care and procedures in rural areas be provided by ambulatory surgery centers close to home. However, the reimbursement rate for such procedures in ambulatory centers is extremely low, so a standard classic intrafascial supracervical hysterectomy procedure needs to be more cost effective to be performed there. Instruments and disposable devices can make up > or = 50% of hospital costs for this procedure, so any cost reduction has to focus on this aspect. METHODS: We identified the 3 most expensive disposable devices: (1) an Endostapler, 498 US dollars and 3 staple reloads, 179 US dollars each; (2) a calibrated uterine resection tool 15 mm for encoring of the endocervical canal, 853 US dollars; and (3) a serrated edged macro morcellator for intraabdominal uterus morcellation, 321 US dollars, and substituted them using classic conservative surgical techniques. RESULTS: From September 2001 to September 2002, we performed 26 procedures with this modified technique at an ambulatory surgery center with a follow-up of 6.7 (2 to 14) months. This modified operative technique was feasible; no conversions were necessary, and no complications occurred. Cost savings were 2209 US dollars per procedure; additional costs were 266.33 US dollars for suture material and an Endopouch, resulting in an overall savings of 50 509.42 US dollars. The disadvantage was an increase in operating room time of about 1 hour 20 minutes per case. CONCLUSION: These modifications in the classic intrafascial supracervical hysterectomy technique have proven to be feasible, safe, and highly cost effective, especially for a rural ambulatory surgery center. Long-term follow-up is necessary to further evaluate these operative modifications. PMID- 15119672 TI - Endokomvos: a simple and secure intracorporeal laparoscopic knot. AB - Accurate placing of securely tied knots in laparoscopic surgery is technically demanding and time consuming. Surgeons must face difficulties arising from 2 dimensional vision, spatial limitations, and restricted movement. Issues to be taken into account include security, virtuosity, and cost effectiveness. The authors believe that in spite of advances in instrumentation and optics, training should aim at manual skill development and application of the basic principles of general surgery. PMID- 15119673 TI - Uncementing relations. PMID- 15119674 TI - Autogenous tooth fragment reattachment--association of periodontal surgery and endodontic and restorative procedures: a case report. AB - The occurrence of dental traumas in adult patients is on the rise, according to the literature, and require correct and adequate treatment specific to each fracture in order to preserve the remaining tooth. In this article, two different fracture cases are reported. Based upon these cases, it may be concluded that there are situations that require the clinician to have a multidisciplinary vision of the different dental specializations in order to effectively treat the patient. PMID- 15119675 TI - Clinical application of all-ceramic fixed partial dentures and crowns. AB - The esthetic demands of patients have increased considerably during recent years. Due to this increasing interest in esthetics, as well as concerns about toxic and allergic reactions, the use of all-ceramic restorations has increased. One of the systems introduced for all-ceramic restorations is the IPS Empress system. This article examines one all-ceramic restoration system, IPS Empress, and presents patient cases in which all-ceramic crowns and fixed partial dentures have been utilized. PMID- 15119676 TI - Surface antibacterial properties of packable resin composites: part I. AB - OBJECTIVE: Restorative materials that possess antibacterial properties are considered advantageous. The aim of this study was to test the antibacterial properties of packable resin composite materials using a direct contact test (DCT) and an agar diffusion test (ADT). METHODS AND MATERIALS: Quadruple samples of SureFil, Alert, P-60, Synergy Compact, Pyramid, and Solitair were placed on the sidewalls of wells in a 96-microtiter plate, and polymerized. A suspension of 10-microL growth medium containing ca. 1 x 10(6) viable mutans streptococci cells were placed on the surface of each sample for 1 hour at 37 degrees C. Fresh medium was then added to each well, and the microtiter plates were placed in a temperature-controlled spectrophotometer which also served as an incubator. Bacterial growth was followed by recording the changes in optical density every 30 minutes for 16 hours. The ADT was performed by placing samples in punched wells of inoculated agar plates, and measurement of the inhibition zone was performed after 72 hours. RESULTS: In both tests, none of the materials inhibited growth of S. mutans. In the DCT, three materials enhanced bacterial growth. Composite samples, aged in phosphate buffered saline for 7 and 30 days, showed no differences when on tested material and controls. CONCLUSIONS: Packable composite materials revealed no antibacterial properties. Furthermore, freshly polymerized composites supported bacterial growth. PMID- 15119677 TI - Evaluation of the sanitization effectiveness of a denture-cleaning product on dentures contaminated with known microbial flora. An in vitro study. AB - OBJECTIVES: To see if dentures contaminated with Staphylococcus aureus, Pseudomas aeruginosa, Bacillus cereus, Candida albicans, and herpes simplex virus 1 could be effectively decontaminated by using Medical Tabs for Dentures. METHOD AND MATERIALS: Ten methylmethacrylate dentures with processed soft liners (soft-liner dentures) and 10 methylmethacrylate dentures without processed soft liners (hard dentures) were aseptically fragmented and individually incubated with a target microorganism. Test denture fragments were immersed in Medical for 5 minutes, vortexed for 5 minutes, and serially diluted onto media. The control denture fragments were similarly treated in sterile water. For virus contamination, denture fragments were contaminated with 1.2 x 10(9) tissue culture infective dose (TCID)50/mL. They were treated with either Medical for 5 minutes (test fragments) or water (controls) for 5 minutes. Serial dilutions were performed and viral (TCID)50/mL titers were calculated using the Reed-Muench method. RESULTS: Medical treatments effectively eliminated C. albicans, S. aureus, and P. aeruginosa from soft-liner dentures. Treatment of hard dentures eradicated C. albicans and reduced the numbers of S. aureus and P. aeruginosa to < 10. B. cereus showed a reduction of 10 microorganisms in hard dentures while the soft liner dentures did not show an appreciable reduction. Viral analyses found that both types of dentures retained large amounts of virus when washed with water, but no virus was recovered from any of the 40 samples treated with Medical. CONCLUSION: A single use of Medical Tabs for Dentures is effective in eliminating certain species of microorganisms, including selected viruses, in vitro. PMID- 15119678 TI - Histologic evaluation of adhesive restorations on dentin caries in rat molar teeth. AB - OBJECTIVE: The aim of this study was to evaluate the progress of dentin caries under resin composite and glass-ionomer cement restorations in vivo. METHOD AND MATERIALS: Sixteen rats were subjected to oral inoculation of the bacterial strain, Streptococcus mutans, and experimental dental caries was induced in the rats' molars. The dental caries induced in rat molars was sealed with resin composite or glass-ionomer cement without removal of the caries; the depth and width of bacterial penetration in the lesion were measured from histopathologically stained sections. Inflammatory cell infiltrations within the pulp were also examined. RESULTS: Both bacterial penetration into dentin and caries spread were significantly reduced by sealing with glass-ionomer cement or resin composite. No significant differences in bacterial penetration and caries spread were seen between the sealing materials. Regarding the pulpal reactions, moderate to severe inflammatory cell infiltration was observed even in the sealed teeth. CONCLUSION: The caries lesions could not be completely arrested by sealing alone, although their progress was slowed from an active to a chronic status. PMID- 15119679 TI - Post-traumatic stress disorder: considerations for dentistry. AB - A dental patient with post-traumatic stress disorder (PTSD) may present with greater dental and behavioral challenges than most dental patients. The background review of PTSD's initiating factors, diagnostic criteria, and medical management should help practitioners better understand and manage these challenges. Many of the challenges the clinician may encounter and managing recommendations are described. A case report of a PTSD patient complaining of constant bilateral tooth pain of the maxillary and mandibular bicuspids and molars is presented. Recommended techniques for identifying the tooth pain source and contributing factors are provided. The primary contributing factor for the patient's tooth pain was determined to be his severe tooth clenching activity. A maxillary acrylic appliance provided some pain reduction and a subsequent mandibular soft occlusal appliance worn opposing the maxillary appliance provided additional relief. PMID- 15119680 TI - Association of malocclusion and functional occlusion with temporomandibular disorders (TMD) in adults: a systematic review of population-based studies. AB - The aim of this systematic review of population-based studies was to establish whether or not associations exist between different types of malocclusions, as well as factors of functional occlusion (eg, occlusal interferences, nonworking side occlusal contacts) and temporomandibular disorders (TMD) in adults 20 years or older. Defined criteria were employed in the search of MEDLINE and EMBASE databases, as well as in a manual search. Finally, using inclusion criteria (eg, random sampling from residents' registration office files or census lists, adequate response rates), out of 22 preselected studies, four relevant population based studies on this subject were found. Eighteen studies were excluded because of insufficient description of material and methods (eg, lack or unclear description of sample method, randomization, age distribution), mixed under- and over-20-year-old study population, or different outcome of interest (eg, tooth loss, dentures). The methodologic quality of the selected studies was established with a quality assessment list. The average total methodologic score achieved was 43 out of a possible 100 points. Few associations were reported between malocclusion and parameters of functional occlusion and clinical as well as subjective TMD, and these associations were not uniform. No particular morphologic or functional occlusal factor became apparent. Additionally, the occlusal factors found were partly protective for TMD, ie, subjects with these occlusal parameters showed fewer signs and symptoms of TMD (angle Class II malocclusion, deep bite, anterior crossbite). A positive relationship was only described in two cases-between the number of rotated lateral teeth and subjective symptoms of dysfunction, and between excessive abrasions and clinical dysfunction. In neither case, however, was the strength of the correlation given. In summary, few associations were established between malocclusion or functional occlusion and signs and symptoms of TMD. In view of the small number of randomized studies and their methodologic quality, these results should be verified through further valid representative studies. PMID- 15119681 TI - Psychobiologic views on stress-related oral ulcers. AB - Recurrent aphthous stomatitis (RAS) and oral lichen planus (OLP), two of the most common stress-related ulcerations of the oral mucosa, have distinct etiologies, courses, and histopathologic features. In light of increased understanding of the psychobiology of stress, the authors propose that RAS and OLP fundamentally differ from the perspective of the psychobiologic response to stress. This article presents possible clinical implications. PMID- 15119682 TI - Osteosarcoma of the maxilla and the maxillary sinus: a case report. AB - Osteosarcoma of the jawbones is a rare malignant mesenchymal neoplasm with the tendency for new bone being directly formed by the tumor cells. Clinically, the tumor may be central or peripheral--periosteal--and histologically can be divided into three subtypes: osteoblastic, fibroblastic, and chondroblastic. This report presents a case of a central osteoblastic osteosarcoma of the left maxillary tuberosity and maxillary sinus. Problems related to definitive diagnosis and therapy are described and discussed. PMID- 15119683 TI - Comparison of autogenous mucosal grafts and collagen-based, solvent-preserved allografts for vestibuloplasty. AB - This study was designed to determine whether solvent-preserved dura mater and fascia lata grafts would be as effective as palatal mucosal grafts for vestibuloplasty. All graft systems succeeded in covering the alveolar ridge with firmly attached tissue, which is needed to ensure prosthetic stability. There was no remarkable reduction in area until the stents were removed. However, after 6 months of vestibuloplasty, there was a significant reduction in gained vestibular area in the fascia lata group, whereas no significant difference between dura mater and palatal grafts was observed. These findings suggest that dura mater could serve as a biologic oral dressing for mucosal defects as effectively as palatal grafts in vestibuloplasty. PMID- 15119684 TI - Use of pedicled buccal fat pad in the closure of oroantral communication: analysis of 75 cases. AB - OBJECTIVE: This report evaluates the use of pedicled buccal fat pad for closure of oroantral communications. METHOD AND MATERIALS: Seventy-five patients were treated with pedicled buccal fat pad. Fifty-two were treated immediately after tooth extractions. The remaining 23 had chronic oroantral communication and were treated similarly after irrigation of the maxillary sinus with saline for 7 days. RESULTS: The 6-month follow-up revealed uneventful healing in all of the patients. Though partial necrosis of the flap was observed in three patients, this did not effect the final healing. Total necrosis of the flap was not noted. CONCLUSION: The use of pedicled buccal fat pad is an acceptable and reliable alternative in acute or chronic oroantral communications management and may even be used as a first treatment choice by experienced surgeons. PMID- 15119685 TI - Basal cell carcinoma. PMID- 15119686 TI - Enhancements to the behavioral parent training paradigm for families of children with ADHD: review and future directions. AB - Behavioral parent training (BPT) is one of the empirically supported psychosocial treatments for ADHD. Over many years and in many studies, BPT has been documented to improve both child ADHD behavior and maladaptive parenting behavior. In some studies, BPT has also been found to result in benefits in additional domains, such as parenting stress and child classroom behavior. However, the BPT literature on children selected as having ADHD lags behind research conducted on BPT for children selected as having oppositional defiant and conduct disorders (ODD and CD, respectively) with regard to examination of factors that may limit treatment attainment, compliance, and outcomes, such as single parenthood, parental psychopathology, and child comorbidity. Because of the high degree of comorbidity between ADHD and ODD/CD, it is difficult to separate the two BPT literatures. The parameters of BPT (e.g.. format and setting), parent factors, and child factors that may contribute to treatment outcomes for families of children with ADHD are reviewed here and recommendations for future BPT research in the area of ADHD are made. PMID- 15119688 TI - A critique of the international consensus statement on ADHD. PMID- 15119689 TI - Paroxysmal postural dyspnea related to multiple large organized thrombi in the left atrium. PMID- 15119687 TI - What are the costs of marital conflict and dissolution to children's physical health? AB - Do parental marital conflict and dissolution influence the risk trajectory of children's physical health risk? This paper reviews evidence addressing this question in the context of understanding how early environmental adversities may trigger a succession of risks that lead to poor health in childhood and greater risk for chronic health problems in adulthood. We first review existing evidence linking marital conflict and dissolution to offspring's physical health outcomes. Next, we provide evidence supporting biopsychosocial pathways that may link marital conflict and dissolution with accelerated health risk trajectories across the lifespan. Specifically, we posit that consequential to the stresses associated with marital conflict and disruption, parenting practices are compromised, leading to offspring deficits in affective, behavioral, and cognitive domains. These deficits, in turn, are hypothesized to increase health risk through poor health behaviors and by altering physiological stress-response systems, including neuroendocrine, cardiovascular, and neurotransmitter functioning. On the basis of the available direct evidence and theoretically plausible pathways, it appears that there is a cost of marital conflict and disruption to children's health; however, more comprehensive investigations are needed to further elucidate this relationship. In the final section, we address limitations in the current literature and identify research that is needed to better evaluate the association between marital conflict and dissolution and children's physical health. PMID- 15119690 TI - Obesity is not only an adult problem. PMID- 15119691 TI - Common ancestors: chronic progressive diseases have the same pathogenesis. AB - In multiple organ systems, chronic progressive disease is characterized at the tissue level by increase in inflammatory cytokines, cell apoptosis and progressive fibrosis, suggesting a possible commonality of pathogenesis. This speculation is supported by the observation that elevated systemic levels of cytokines and/or CRP predict the appearance of disease, progression of disease, and disease complications. Some therapeutic interventions that reduce the expression of inflammatory cytokines, such as HMG Co-A reductase inhibitors, slow the progression of a number of chronic diseases, independent of their effect on cholesterol level. Taken together, these data suggest a unifying hypothesis for many chronic progressive diseases. Diverse noxious stimuli activate a normal protective inflammatory response. This response typically defervesces with elimination of the stimulus, reestablishing homeostasis. When homeostasis is not restored, that is, the inflammatory response persists, cell apoptosis and tissue fibrosis can result. The universality of this tissue response derives from the fact that all cells derive from the same blastocyst. Since cell differentiation results in different tissue functions, however, the clinical manifestations of disease are also vastly different, obscuring the underlying disease process. The relevance of this insight is that it provides a potential framework for testing old and new therapies which might inhibit a diverse set of clinical conditions at several levels of the disease process. PMID- 15119692 TI - Role of noninvasive studies in risk stratification for sudden cardiac death. AB - Sudden cardiac death (SCD) is one of the leading causes of mortality in developing countries. To prevent SCD, it is crucial to have effective tools for identifying patients at risk, given that there are now effective devices and medications that can prevent SCD. Two noninvasive electrocardiographic tools have been approved by the Food and Drug Administration and are being used clinically for identifying patients at risk for SCD: the signal-averaged electrocardiogram (SAECG) and T-wave alternans (TWA). This article reviews each of these approaches and includes a discussion of the mechanisms and a summary of the clinical studies published to date. PMID- 15119693 TI - Effects of conventional and aggressive statin treatment on markers of endothelial function and inflammation. AB - BACKGROUND: Atherosclerosis is considered to be a chronic inflammatory disorder. Several large-scale clinical studies demonstrate that markers of inflammation, such as high-sensitivity C-reactive protein (hsCRP), fibrinogen, and soluble CD40 ligand, are potent and independent predictors of vascular risk. HYPOTHESIS: The study was undertaken to investigate the effect of increasing the statin dose from conventional to aggressive treatment on lipids levels, inflammation, and endothelial function in patients with coronary artery disease (CAD). METHODS: We randomized 97 patients to either 20 mg simvastatin or 80 mg atorvastatin. Plasma levels of lipids, hsCRP, fibrinogen, soluble adhesion molecules, and nitric oxide total were analyzed at baseline and after 6 months of treatment. RESULTS: Lipid values were significantly reduced in both treatment groups, but with significantly greater reduction in the aggressively treated group. Furthermore, aggressive statin treatment significantly decreased hsCRP and fibrinogen, while only small reductions were seen in the conventionally treated group, resulting in significant differences between the two treatment groups (p < 0.001). Nitric oxide-total increased significantly in both treatment groups, although the increase was more pronounced in the aggressively treated group (22.6 vs. 15.6%). CONCLUSION: Aggressive statin treatment significantly improved lipid status and reduced markers of inflammation and improved endothelial function compared with conventional treatment in patients with CAD. No interaction was observed, and high-dose treatment did not offer additional benefit compared with standard-dose treatment with respect to soluble adhesion molecules. PMID- 15119694 TI - Dynamic 123I-BMIPP single-photon emission computed tomography in patients with congestive heart failure: effect of angiotensin II type-1 receptor blockade. AB - BACKGROUND: Heart failure is a major and growing public health problem with a high mortality rate. Although recent studies have demonstrated that a variety of metabolic and/or neurohumoral factors are involved in the progression of this syndrome, the precise mechanisms responsible for this complex condition are poorly understood. HYPOTHESIS: To examine 123I-beta-methyl iodophenylpentadecanoic acid (BMIPP) kinetics in the early phase soon after tracer injection in patients with congestive heart failure (CHF), we performed dynamic single-photon emission computed tomography (SPECT). METHODS: Twenty-six patients with CHF and eight control subjects were examined. The consecutive 15 images of 2-min dynamic SPECT were acquired for 30 min after injection. In the early phase after injection (0-4 min), a significant amount of radioactivity existed in the blood pool. After 6 min, the myocardial 123I-BMIPP image was clear and thus the washout rate of 123I-BMIPP from 6 to 30 min was calculated. RESULTS: The washout rate of 123I-BMIPP from the myocardium was faster in patients with CHF than in the controls (8 +/- 4 vs. -5 +/- 3%, p < 0.01). The washout rate of 123I-BMIPP demonstrated positive correlation with left ventricular (LV) end diastolic volume index (R = 0.54, p < 0.02) and inverse correlation with LV ejection fraction (R = 0.53, p <0.02). Patients were given the angiotensin II type-1 receptor antagonist candesartan for 6 months, and dynamic SPECT was repeated. The enhanced washout rate of 123I-BMIPP in CHF was reduced after treatment with candesartan (p < 0.05). CONCLUSION: These data suggest that (1) enhanced washout of 123I-BMIPP was observed soon after injection in patients with CHF, (2) the activation of angiotensin II signaling pathway is involved as an intracellular mechanism for enhanced 123I-BMIPP washout in heart failure, and (3) improvement in fatty acid metabolism may represent a new mechanism for beneficial effects of angiotensin II receptor blockade on cardiac function and survival in patients with heart failure. 123I-BMIPP washout in the early phase obtained from dynamic SPECT may be a new marker for evaluating the severity of heart failure and the effects of medical treatment. PMID- 15119696 TI - Images in cardiology: Unusual anomalies of the left circumflex artery. PMID- 15119695 TI - C-reactive protein plasma levels but not factor VII activity predict clinical outcome in patients undergoing elective coronary intervention. AB - BACKGROUND: Both vascular inflammation as determined by C-reactive protein (CRP) and extrinsic coagulation as measured by factor VII activity (F VII) may predict clinical restenosis rate in patients with stable angina pectoris undergoing elective percutaneous coronary intervention (PCI). HYPOTHESIS: The primary objective of this study was to investigate the associations between baseline CRP levels, F VII activity, and restenosis rate after elective PCI in a 6-month follow-up period. METHODS: This prospective study included 81 patients aged > or = 19 years undergoing PCI for angiographically significant (> or = 70%) stenosis, with or without stenting, and 49 controls. Factor VII activity and CRP were measured in samples collected at angiography and 16-24 h post procedure after overnight fast. Successful PCI was defined as final diameter of < 50% with TIMI 3 flow and no complication within 1 h. After 6 months all patients who had undergone PCI were evaluated via a standardized questionnaire. Clinical restenosis was defined as the occurrence of a major adverse coronary events (MACE), within the follow-up period. RESULTS: Diagnostic angiography led to a significant increase in CRP levels after 16-20 h in patients with discrete CAD (n = 22) but not in patients without any signs of coronary atherosclerosis (n = 27). During a 6-month follow-up after PCI, 17 of 81 (21%) patients developed MACE. Tertiles of CRP levels independently predicted clinical restenosis, as it developed in 33.3% of patients with the highest CRP levels (0.7-4.8 mg/dl), in 16.6% of patients with second tertile CRP levels (0.23-0.69 mg/dl), and in 7.4% of patients with lowest tertile CRP levels (0.0-0.22 mg/dl). There was a significant difference in the restenosis rate between patients from the first and the third tertiles (p = 0.018). Successful PCI was associated with a significant decrease of mean CRP levels after 6 months, whereas PCI in patients suffering from MACE led to no change in CRP levels. There was no association between factor VII activity and clinical outcome after PCI, and F VII activity did not change over a 6-month period. CONCLUSIONS: In patients with stable angina pectoris undergoing elective PCI, increased preprocedural and 6-month follow-up CRP plasma levels are associated with clinical restenosis. Factor VII plasma activity lacks such correlations. PMID- 15119697 TI - Ventricular tachyarrhythmia associated with cardiac sarcoidosis: its mechanisms and outcome. AB - BACKGROUND: Cardiac sarcoidosis is increasingly recognized and is associated with poor prognosis. Ventricular tachycardia (VT) associated with cardiac sarcoidosis is the most likely cause of sudden death in most patients, but the mechanism has not been well established. HYPOTHESIS: This study investigated the mechanisms and outcome of VT associated with cardiac sarcoidosis. METHODS: The study included eight consecutive patients (five men, three women, aged 54 +/- 19 years) who had sustained monomorphic VT associated with cardiac sarcoidosis in our hospital. RESULTS: The average ejection fraction was 43 +/- 11%. Twenty-two VTs were observed in these patients, and mean heart rate during VT was 192 +/- 29 beats/min (range 144-259). The phenomenon of transient entrainment was documented in 10 of 22 (45%) VTs by ventricular pacing (eight in the active phase). Another five (23%) VTs could not be entrained, but could be initiated by programmed stimulation and terminated by rapid pacing, reproducibly. In 3 of the 22 (14%) VTs, cardioversion was required urgently because of the fast rate, while the remaining 4 (18%) could be induced during electrophysiologic study. CONCLUSIONS: In this study, there was a high possibility that the mechanism of 15 (68%) VTs was reentry. Reentrant substrate is formed not only in association with the healing of cardiac granulomas in the inactive phase of cardiac sarcoidosis but also in the active phase. Ventricular tachycardia with cardiac sarcoidosis, even if this mechanism is reentry, has different inducibility between the active and inactive phases in an electrophysiologic study. This makes the therapy for cardiac sarcoidosis (e.g., corticosteroids, antiarrhythmic agents, and catheter ablation) difficult. The implantable cardioverter-defibrillator is an effective treatment for ventricular tachyarrythmia with cardiac sarcoidosis. PMID- 15119698 TI - Images in cardiology: Accidental perforation of the left ventricle during angiography. PMID- 15119699 TI - Effect of chronic stress and sleep deprivation on both flow-mediated dilation in the brachial artery and the intracellular magnesium level in humans. AB - BACKGROUND: Chronic mental and physical stress has been suggested to be a trigger for cardiovascular events. In addition, a reduction in levels of intracellular magnesium has been reported to cause vasoconstriction while enhancing platelet dependent thrombosis. HYPOTHESIS: The purpose of this study was to investigate whether chronic stress affects endothelial function and intracellular magnesium levels in humans. METHODS: Flow-mediated dilation (endothelium-dependent vasodilation) and sublingual nitroglycerin-induced dilation (0.3 mg, endothelium independent vasodilation) were measured in the brachial artery in 30 healthy male college students, aged 22 +/- 1 years, using high-resolution ultrasound both before and immediately after a 4-week final term examination period. Erythrocyte magnesium concentration was measured simultaneously. All students had chronic sleep deprivation for 4 weeks, during which sleep lasted < 80% of that on ordinary days; in addition, the students were under great stress to pass the examination. This condition was considered to be chronic stress. RESULTS: Chronic stress decreased flow-mediated dilation and erythrocyte magnesium concentration (from 7.4 +/- 3.0 to 3.7 +/- 2.3%, p < 0.05; from 5.7 +/- 0.4 to 5.5 +/- 0.4 mg/ml, p < 0.05, respectively). The change in flow-mediated dilation correlated significantly with that of the erythrocyte magnesium concentration (r = 0.43, p < 0.05), but not with nitroglycerin-induced dilation. CONCLUSIONS: Chronic stress was found to attenuate endothelial function, which may also be associated with a reduction in the intracellular magnesium level in humans. PMID- 15119700 TI - Dynamic assessment of myocardial involvement in patients with end-stage renal disease by ultrasonic tissue characterization and serum markers of collagen metabolism. AB - BACKGROUND: Congestive heart failure is the most common cause of mortality in patients with end-stage renal disease (ESRD). However, noninvasive assessment for cardiac involvement in ESRD has not been established. HYPOTHESIS: Assessment of ultrasonic tissue characterization and serum markers of collagen degradation is useful for defining myocardial involvement in ESRD. METHODS: Cyclic variation of ultrasonic integrated backscatter of the ventricular septum (CV-IBS) and the serum levels of free matrix metalloproteinase-I (MMP-I) and tissue inhibitor of metalloproteinase-I (TIMP-I) were measured in 30 patients with ESRD undergoing routine hemodialysis (HD) and in 40 patients with essential hypertension (HTN). RESULTS: Compared with the group with HTN, ESRD (before HD) showed larger left ventricular (LV) mass index (217 +/- 56 vs. 146 +/- 45 g/m2, p < 0.01), worse LV diastolic function (E/A, 0.6 +/- 0.2 vs. 0.9 +/- 0.3, p < 0.05), smaller CV-IBS (9.0 +/- 1.3 vs. 12.4 +/- 0.9 dB, p < 0.01), and larger TIMP-I/MMP-I (46 +/- 10 vs. 34 +/- 10, p < 0.05), in spite of the comparable ventricular wall thickness. Thus, these indices may possibly reflect myocardial interstitial fibrosis. After HD (after the improvement of myocardial interstitial edema), a negative linear relationship between CV-IBS and TIMP-I/MMP-I was observed (r= -0.52, p < 0.05). CONCLUSIONS: Noninvasive assessment of ultrasonic tissue characterization and serum markers of collagen type I degradation may be a new diagnostic tool for defining myocardial interstitial fibrosis in patients with ESRD and LV hypertrophy. PMID- 15119701 TI - Patterns of congenital heart disease in unoperated adults: a 20-year experience in a developing country. AB - BACKGROUND: The number of patients with congenital cardiac disease reaching adulthood is increasing steadily. Many adults with such disease face both medical and surgical difficulties. HYPOTHESIS: This retrospective study was undertaken to assess the frequency and outcome of congenital heart disease (CHD) in unoperated adults. METHODS: The charts of all patients with unoperated CHD, who were admitted to a tertiary care center in Beirut, Lebanon, between 1980 and 2000 were reviewed. Of these, 206 patients (52% men, age at admittance 18-71 years [32.8 +/ 13.3 years]) with a diagnosis of CHD were evaluated. Atrial septal defect (ASD) was the most common cardiac malformation with a relative frequency of 53%, followed by ventricular septal defects (11%), tetralogy of Fallot (11%), aortic anomalies (7%), pulmonary stenosis (6%), and Ebstein anomaly (4%). Most patients were symptomatic upon presentation, with dyspnea on exertion being the most common presenting symptom. Twenty-seven patients (13%) had cyanotic CHD. Of 179 acyanotic patients, 113 (63%), and 17 of 27 cyanotic patients (63%) underwent surgical intervention. In-hospital surgical complications for the acyanotic group included cerebrovascular accident (2%) and heart block (1%). Total surgical mortality was 4 of 130 (3%). One patient with tetralogy of Fallot presented with endocarditis and died. CONCLUSION: Atrial septal defect is the most common defect reported in our experience; however, it occurs more frequently than that reported in the literature. Although most patients were symptomatic on presentation, their functional status was stable. Accordingly, their hospital course, whether managed medically or surgically, held a relatively low complication rate. This could be attributed to the uncomplicated nature of pathologies in our series. The surgical mortality and in-hospital complications were slighter higher than those reported for similar lesions if repaired during childhood. This study reflects the relative frequency of various cardiac malformations in selected patients with "grown up" congenital heart disease (GUCH) and their natural survival pattern. PMID- 15119702 TI - Rapid screening of cardiac patients with a miniaturized hand-held ultrasound imager--comparisons with physical examination and conventional two-dimensional echocardiography. AB - BACKGROUND: Rapid screening of cardiac patients with a hand-held ultrasound imager (SonoHeart [SH]) could provide valuable clinical information. HYPOTHESIS: Whether the use of this device yields additional information to a carefully conducted physical examination and comparable findings to those of conventional two-dimensional echocardiography (2-D) during inpatient rounds is not well established and is the subject of this study. METHODS: In all, 100 consecutive telemetry patients underwent rapid screening with 2-D and color Doppler SH during inpatient rounds. SonoHeart findings were compared with results from conventional 2-D and physical examination conducted by an attending cardiologist. RESULTS: All patients had interpretable images. Mean scanning time with SH was 5.0 +/- 1.2 min; 2-D and SH findings were comparable. The parameters studied included chamber sizes, left ventricular (LV) systolic function, presence of LV hypertrophy (LVH), wall motion abnormalities (WMA), pericardial effusion (PE), and valvular regurgitations. Mild to moderate valvular regurgitation and LV systolic dysfunction were reliably diagnosed by SH in a number of patients whose symptoms were unrelated to the abnormalities detected. CONCLUSIONS: Rapid screening with SH provides accurate and valuable information that would otherwise be undetected during physical examination. Its introduction into clinical practice may redefine the initial approach to patients with cardiovascular disease. PMID- 15119703 TI - Images in cardiology: Radiographic appearance of a left ventricular apical aneurysm with thrombus following a large anterior myocardial infarction: "dumbbell" shaped heart. PMID- 15119704 TI - Edmund H. Sonnenblick. PMID- 15119705 TI - [Restoration of Class V cavities with the Ormocer-based filling system Admira]. AB - The aim of this study was to evaluate the clinical effectiveness of an Ormocer based restorative system for treatment of mixed carious and non-carious class V cavities. 70 restorations in 25 patients were placed with the condensable material Admira (group 1). In a second group of eight patients, the treatment of 25 cavities took place with the flowable material Admira Flow in combination with composite-bonded to-flowable technique or with the flowable material alone. The clinical evaluation was carried out using a modified clinical criteria system (Bohm et al. 1991) after Lutz et al. (1977) and Ryge (1980). Restorations of group 1 reached the two-year-level, those of group 2 the one-year-level. Good results were obtained with both condensable material and flowable material despite the small number of failures in the various clinical criteria. The retention rate of the flowable composite was 100% after six and twelve months and was the only parameter superior to those of the condensable material (98.6% resp. 93.9%). After two years, group 1 showed significant differences to base line with respect to volume behaviour and surface texture. No significant differences were determined with respect to of colour match, bond zone morphology and clinical acceptance. A SEM evaluation of replicas confirmed clinical results but also revealed clinically invisible failures of bond zone morphology. PMID- 15119706 TI - [Survival rate of IPS-Empress 2 all-ceramic crowns and bridges: three year's results]. AB - The objective of this prospective clinical study was to calculate the survival rate of IPS-Empress2 crowns and fixed partial dentures (FPD) over a three-year period. In 43 patients 27 IPS-Empress2 crowns and 31 fixed partial dentures were adhesively luted. Crowns were placed on premolars and molars and FPDs were inserted in the anterior and premolar area. Abutments were prepared with a circular 1.2 mm wide shoulder. The clinical follow-up examination took place after 6, 12, 24, 36 and 48 months. After a mean of 38 months, the survival rate (Kaplan-Meier) of all-ceramic crowns was 100% and of the three unit FDP 72.4%. There were a total of six complete failures which occurred only with the three unit IPS-Empress2 FPDs. Three FPDs exhibited fractures of the framework for which the manufacturer's instructions of connector-dimension was not satisfied, and one FPD exhibited an irreparable incomplete veneer fracture. Further two FPDs showed biological failures. The accuracy of fit and esthetics were clinically satisfactory. The three-year results showed the IPS-Empress2-ceramic as an adequate all-ceramic material for single crowns. The use for FPD needs further critical consideration. PMID- 15119707 TI - [Limits of total prostheses in resected patients. Case report]. AB - The prosthetic treatment of patients with oral defects after tumor resection by means of complete dentures is demanding and difficult. In the past various surgical procedures were performed for corrections of the alveolar ridges and defects, with unpredictable results and poor prognosis. The intraoral situation often resulted in difficulties of wearing complete dentures. Nowadays oral implants offer better treatment possibilities to these patients with compromised oral structures. Due to the specific anatomical conditions standardized treatment modalities and procedures are sparse and not documented. Individual treatment planning along with the use of implants may favor a positive treatment outcome of the prosthetic reconstruction, providing satisfactory function. On the basis of a case report the authors present the implant prosthodontic reconstruction of a patient who was unable to wear a complete denture after a partial resection of the maxilla due to an oral tumor. PMID- 15119708 TI - [Indications for use of MTA, a review. Part 1: Chemical, physical and biological properties of MTA]. AB - Mineral trioxide aggregate is a recently introduced dental material. It may be used to seal perforations, make retrograde fillings in root-end resections, seal open apices, or cap vital pulps. In this review of the literature, physical, chemical, and biologic properties of MTA are discussed. Studies have suggested that MTA provides a better seal than formerly used materials such as IRM, amalgam, and Super-EBA. Further, MTA has low cytotoxicity and excellent biocompatibility. In vivo studies demonstrated a beneficial effect of MTA on pulpal and periodontal regeneration. Although controlled randomized clinical trials are still missing, MTA appears to be a suitable material to tightly seal dental hard tissues from the periodontium, or to cap the exposed pulp. PMID- 15119709 TI - Advances in implant dentistry. PMID- 15119710 TI - Intracanal reinforcement fiber in pediatric dentistry: a case report. AB - A technique for the restoration of carious primary maxillary incisors using indirect resin composite crowns and intracanal reinforcement fiber is described. Endodontic treatment was previously performed on each tooth. The advantages of using an intracanal reinforcement fiber include resin composite crown reinforcement, translucency, and relative manipulation facility. In addition, the use of indirect resin composite crowns provides good shape and esthetics, as well as reduced chair time for the child. The technique is illustrated in a case report in which indirect resin composite crowns and an intracanal reinforcement fiber are placed in a 3-year-old girl. PMID- 15119711 TI - Comparison of laboratory and clinical wear rates of resin composites. AB - OBJECTIVE: The purpose of this study was to use wear simulation to develop wear rates for two modern composite systems and then compare these rates with clinical studies on the same materials. METHOD AND MATERIALS: A spring-loaded piston wear simulator was used to generate localized wear rates for P50 and Z100 at 100,000, 200,000, 300,000, and 400,000 cycles. Clinical studies on P50 and Z100 conducted at Creighton University and the Catholic University of Leuven were used for comparison of clinical wear to the laboratory values. Regression analysis was employed to define the wear rates. RESULTS: The laboratory wear rates determined with simulation for P50 and Z100 were similar. Clinical occlusal contact area (OCA) wear for P50 and Z100 had small differences and were pooled to provide comparison with laboratory data. Wear rates determined from linear regression analysis provided equivalency factors between laboratory localized wear and clinical OCA wear that could be used for future studies. Further regression analysis comparing OCA and generalized clinical wear allowed a conversion factor of 4.5 to be determined. CONCLUSION: Localized wear from laboratory simulation may be a useful predictor of localized clinical wear and also clinical generalized wear using a conversion factor. PMID- 15119712 TI - Antibacterial surface properties of polymerized single-bottle bonding agents: part II. AB - OBJECTIVE: Microorganisms are directly associated with the etiology of enamel, dentin, and pulpal pathology. Due to the growing usage of one-bottle bonding materials with resin composite restorations, as well as sealing agents with amalgam restorations, it is important that they possess antibacterial properties. In the present study, the antibacterial properties of polymerized one-bottle bonding agents were tested, using the direct contact test (DCT) and the agar diffusion test (ADT). METHOD AND MATERIALS: Quadruple samples of the following materials were tested in both assays: Bond-1, OptiBond Solo, One-Step, Gluma, Prime & Bond NT, and Synergy. In the DCT, samples were placed on the sidewalls of wells of a 96-microtiter plate and polymerized. A 10-microL suspension of Streptococcus mutans was placed on the surface of each sample for 1 hour at 37 degrees C. Fresh media was then added, and bacterial growth was followed with a temperature-controlled spectrophotometer. In the ADT, samples were placed in punched wells of inoculated agar plates, and halos in the bacterial lawn were measured after 72 hours. RESULTS: In the DCT, all the tested bonding agents exhibited potent antibacterial properties; virtually no viable bacteria were present in any of the samples. When the samples were aged in phosphate-buffered saline for 24 hours, all the tested agents, similar to the freshly polymerized samples, exhibited potent antibacterial properties. This property was lost in samples aged for 7 days. Fresh samples assayed by ADT demonstrated no inhibition halo around any of the samples. CONCLUSION: Collectively, the present data suggest that one-bottle bonding agents possess in vitro antibacterial properties for at least 24 hours. This phenomenon was demonstrated only by DCT. PMID- 15119713 TI - Fracture resistance of teeth restored with indirect-composite and ceramic inlay systems. AB - Fracture resistance of dentin-bonded inlays may be influenced by the restorative material used. The purpose of this study was to evaluate, in vitro, the fracture resistance of teeth restored with four tooth-colored materials: feldspathic ceramic, Duceram LFC, and three laboratory resins, Solidex, Artglass, and Targis. Sixty mandibular molar teeth were placed in resin cylinders, reproducing the periodontal ligaments. Mesial-occlusal-distal (MOD) inlay preparations were made in a standard cavity preparation appliance. Subsequently, the teeth were molded, and the restorations were prepared following the manufacturer's instructions. The inlays were cemented with resin composite cement, Rely X, and stored at 37 degrees C and 100% humidity for 24 hours. The samples were then submitted to an axial compression load at a speed of 0.5 mm/minute. Statistical analysis by one way ANOVA and Tukey's multiple comparison test revealed that the teeth restored with Duceram LFC (205.44 +/- 39.51 kgf) showed statistically inferior fracture resistance than the three other groups restored with indirect resin composites (Solidex [293.16 +/- 45.86 kgf], Artglass [299.87 +/- 41.08 kgf], and Targis [304.23 +/- 52.52 kgf]). PMID- 15119714 TI - Convergence angle, occlusal reduction, and finish line depth of full-crown preparations made by dental students. AB - OBJECTIVE: To measure the convergence angle, incisal/occlusal clearance, and finish line depth, which indicate the amount of axial reduction of full metal ceramic crown preparations made by final-year dental students. METHOD AND MATERIALS: A total of 157 preparations were assessed. Convergence angle and finish line depths were measured by Tool Maker Microscope, while crown height was measured using a digital caliber. RESULTS: The mean convergence angle, faciolingually and mesiodistally, was 22.4 and 25.3 degrees, respectively. The mean for molars was statistically significantly greater than that for other teeth. The incisal/occlusal reduction ranged from 1.8 to 2.2 mm and averaged 2 mm. Buccal shoulder finish line depth averaged 0.86 mm, and mean chamfer finish line depth was 0.74 mm lingually, 0.7 mm mesially, and 0.66 mm distally. The buccal shoulder for mandibular anterior teeth was significantly smaller than for other teeth and averaged only 0.45 mm. CONCLUSION: The mean convergence angle was clinically acceptable. However, shoulder depth was less than the recommended depth of 1.0 to 1.5 mm for metal ceramic crowns. Incisal/occlusal reduction was clinically adequate to provide enough structural durability and color matching. PMID- 15119715 TI - A three-visit, complete-denture technique utilizing visible light-cured resin for tray and base plate construction. AB - This article describes a technique of constructing a set of maxillary and mandibular complete dentures in three visits instead of the usual five clinical appointments. This system of complete-denture construction is made possible because of the combined use of visible light-cured material as an impression tray and record base material, as well as the use of new biometric wax occlusion rims. Unlike some earlier techniques that use light-cured resin composites as the denture base materials, this method retains the use of heat-cured polymethylmethacrylate as the denture base material. PMID- 15119717 TI - Air entrapment in the cervicofacial region due to injury. AB - Air entrapment in the soft tissues of the cervicofacial region is a benign entity occasionally developing after a fracture of the facial bones or ribs. Characteristic clinical findings of air entrapment are the sudden onset of edema and crepitation of the respective region. Possible complications are respiratory obstruction or contamination of the mediastinum. Treatment includes precautionary administration of antibiotics. In regard to the region of injury, air may enter the anterior cranial fossa where it compresses the brain, resulting in pneumocephalus, which may cause meningitis as an imminent complication. After a fracture at the zygomatic or nasal bones, subcutaneous emphysema may result in the middle third of the face. It is also possible for air to penetrate into the retrobulbar fat and damage the eye. In other cases, air traverses the parapharyngeal space and causes pneumomediastinum. Moreover, after a fracture at the mandible, it is possible that air passes through the floor of the mouth and the submandibular area to the deep tissue layers again causing pneumomediastinum. Alternatively, after rib fracture, air could follow an upward course, resulting in subcutaneous emphysema of the thoracocervicofacial region. In this article, patients who suffered from air entrapment were categorized into four groups, and the methods used to diagnosis and treat these patients are presented. PMID- 15119716 TI - Effects of incremental curing on contraction stresses associated with various resin composite buildups. AB - OBJECTIVE: The purpose of this study was to photoelastically evaluate contraction stresses associated with various resin composite build-up procedures, including incremental curing and the use of flowable composite for pulpless molars. METHOD AND MATERIALS: Life-sized photoelastic models of an endodontically treated molar were fabricated. The cavity represented a conservative access preparation and included four lateral walls. The following materials were used for buildup: dual cured hybrid composite (Cleafil DC Core [DC], BIS-CORE [BC]); light-cured flowable composite (AELITEFLO LV [ALV]); and chemical-cured flowable composite (CORE-FLO [CF]). The photoelastic models were built up with the following techniques: bulk-cured (BDd [dual-cured DC], BDc [chemically-cured DC], BBd [dual cured BC], and BC [CF]); and incremental-cured (1 mm gingivally + 3.5 mm occlusally; IAB [ALV + BC], ICB [CF + BC], and IBB [BC + BC]). Isochromatic fringes developed in the models were recorded photographically in the field of a circular polariscope, and maximum fringe order was determined. Five specimens were tested for each condition. RESULTS: Stress intensity of the build-up methods fell into two categories: high (BDd, BBd, BC, and IAB) and low (BDc, ICB, and IBB). The difference between high and low groups was statistically significant, except between BC and ICB. The maximum fringe order was developed around the point angles at the cavity floor for all the conditions tested. CONCLUSIONS: Type and setting mechanism of resin composite build-up technique had considerable influence on contraction stress. Incremental buildup using composites with low elastic modulus did not reduce contraction stress intensity compared with bulk cured techniques. PMID- 15119719 TI - Schizophrenia and dental management: review of the literature. AB - Schizophrenia is a brain disease that manifests itself with multiple signs and symptoms involving thought, perception, emotion, and behavior. Despite the fact that these manifestations combine in various ways to create considerable diversity among patients, the cumulative effect of the illness is always severe and usually long lasting. Schizophrenia is diagnosed in the presence of certain positive and negative psychotic symptoms, as well as psychologic and cognitive deterioration. In addition to history, medical and psychiatric examinations are used to rule out other possible diagnoses. Schizophrenia is not a rare disorder, and it affects approximately 1% of the general population. Many of these individuals require several hospitalizations, and the financial cost of schizophrenia is substantial. In this article, clinical features of schizophrenia are reviewed, and the important consequences from the perspective of dental practitioners are highlighted. PMID- 15119718 TI - Infection-related inferior alveolar and mental nerve paresthesia: case reports. AB - Nerve injury can be related to mechanical, chemical, and thermal factors. Infection-related paresthesia is usually related to mechanical pressure and ischemia associated with the inflammatory process. Another cause of paresthesia could be the toxic metabolic products of bacteria or inflammatory products released following tissue damage. This article presents cases of inferior alveolar and mental nerve paresthesia caused by an infected impacted tooth, an infected cyst, and periapical infection. The possible pathophysiologic mechanism of nerve injury, therapy, and prognosis for recovery are also discussed. PMID- 15119720 TI - Bitewing film quality: a clinical comparison of the loop vs. holder techniques. AB - OBJECTIVE: To compare in vivo bitewing film quality using the holder versus the paper loop technique. METHOD AND MATERIALS: Four bitewing films were taken from the right and left premolar and molar regions of 45 dental students using both the bitewing holder and paper loop techniques. A total of 360 films were taken and assessed by an experienced practitioner not apprised of the bitewing technique used. Of interest were: (1) the number of overlaps and the percentage of teeth showing the alveolar crest; (2) proper film positioning; and (3) the percentage of cone cutting. A Poisson regression using generalized estimating equations (GEEs) was used to estimate the difference in overlap between the two techniques. For proper positioning and cone cutting, logistic regressions using GEEs were used. RESULTS: The average number of horizontal overlaps for the loop and holder techniques at the right premolar, right molar, left premolar, and left molar were 1.64, 2.11, 2.16, 2.78, and 1.64, 2.00, 2.00, 2.18, respectively. The loop technique was 1.11 times more likely to cause overlapping than the holder technique. The highest percentage of teeth showing the alveolar crest by the loop technique was 97.8% in the mandibular second premolar and first molar. With respect to film positioning, the loop technique was 1.12 times more likely to cause improper positioning than the holder technique. Both techniques demonstrated minimal cone cutting (1 in the loop versus 0 in the holder). CONCLUSION: The quality of bitewing films taken by the loop and holder techniques was not significantly different. PMID- 15119721 TI - The long-term sealing ability of AH 26 and AH plus used with three gutta-percha obturation techniques. AB - OBJECTIVE: To evaluate and compare the sealability of root fillings in extracted teeth by using AH 26 and AH Plus in conjunction with three different obturation techniques. METHOD AND MATERIALS: Root canals of 940 single-rooted teeth were prepared according to the crown-down/stepback technique (using both 2.5% NaOCl and File-Eze) before lateral condensation and hybrid condensation of gutta percha or obturation with Thermafil. Teeth were immersed in India ink for 90 hours after storage for 1 day, 1 week, 2 weeks, 1 month, and 6 months. The roots were split longitudinally, and the extent of dye penetration was measured using a stereomicroscope. Statistical analysis was performed using Kruskal-Wallis test and Mann-Whitney U test. RESULTS: There were no statistically significant differences in apical leakage within the AH 26 and the AH Plus groups, nor between the two sealer groups, at any observation period. Coronal leakage was significantly higher for Thermafil compared to hybrid condensation at 1 day, 1 week, and 2 weeks both for AH 26 and AH Plus. Coronal leakage was higher only for Thermafil compared to lateral condensation at 1 week for AH 26 and for lateral condensation compared to hybrid condensation at 1 week for AH Plus. CONCLUSION: Both AH 26 and AH Plus, when used with an identical gutta-percha obturation technique, resulted in comparable sealability at all evaluation times and in comparable coronal sealability at 1 and 6 months. PMID- 15119722 TI - Caries prevention in pediatrics: dietary guidelines. AB - Establishment of good dietary habits during infancy and childhood can minimize risk of caries development throughout life. Optimal dietary habits for oral health are consistent with dietary recommendations for systemic health, growth, and development. Minimally processed foods should be provided at defined meals and snacks. PMID- 15119723 TI - Amelogenesis imperfecta. PMID- 15119725 TI - Vet to be struck off for failing to provide adequate professional care. PMID- 15119724 TI - Government rejects FAWC's recommendations on religious slaughter. PMID- 15119726 TI - 'Wake-up call' on veterinary nursing... PMID- 15119727 TI - Who will regulate the veterinary profession of the future? PMID- 15119728 TI - Vaccination in cats: which ones, and how often? PMID- 15119729 TI - Serum antibody titres to canine parvovirus, adenovirus and distemper virus in dogs in the UK which had not been vaccinated for at least three years. AB - Antibody titres to canine distemper (CDV), canine parvovirus (CPV) and canine adenovirus (CAV) were measured in 144 adult dogs that had not been vaccinated for between three and 15 years. Protective antibodies to CPV were present in 95 per cent of the population, to CDV in 71.5 per cent and to CAV in 82 per cent. The prevalence of protective titres did not decrease with increasing time interval from the last vaccination for any of the three diseases studied. Booster vaccination increased the dogs CAV titres. For comparative purposes, 199 puppies were sampled at the time of their first and second vaccination. In the case of CPV and CAV a significantly higher proportion of the adult dogs were protected than of the puppies immediately after they were vaccinated. Natural CPV boosting was strongly suspected because the dogs had significantly higher titres three years after their primary vaccination than two weeks after it and three unvaccinated dogs had acquired protective antibody levels uneventfully. There was no evidence of natural exposure to CDV. PMID- 15119730 TI - Indirect transmission of bovine viral diarrhoea virus at calving and during the postparturient period. AB - Two trials were designed to investigate whether bovine viral diarrhoea virus (BVDV) could be transmitted after the birth of persistently infected calves, even if they were removed immediately after birth. In trial 1, 11 calves were actively exposed to fetal fluids and uterine lochia collected from cows that had delivered calves persistently infected with type 1 BVDV. One calf that was exposed to a sample taken on the day of calving seroconverted. In trial 2, six calves were housed in stables where persistently infected calves were being born and then removed immediately from their dams and from the stable unit within two to three hours. One of four calves in close contact with the cows after delivery seroconverted and one of two calves housed within the same stable unit became infected. PMID- 15119731 TI - Trombidiosis in cats caused by the bite of the larval trombiculid mite Helenicula miyagawai (Acari: Trombiculidae). PMID- 15119732 TI - Efficacy of seven disinfectant sanitisers on field isolates of Brachyspira pilosicoli. PMID- 15119733 TI - Subcutaneous fibrous hamartoma in a foal. PMID- 15119734 TI - Serological survey for leptospires of agricultural importance in common wombats (Vombatus ursinus). PMID- 15119735 TI - Ovine gastrointestinal listeriosis. PMID- 15119736 TI - Suspected necrotic enteritis in wild swans. PMID- 15119737 TI - Vetting spelling. PMID- 15119738 TI - Amyloid precursor protein gene analysis in familial Alzheimer's disease cases: a lack of mutations in exons 16 and 17. AB - The beta-amyloid precursor protein (APP) gene (on chromosome 21), Presenilin 1 (PS1) gene (on chromosome 14) and Presenilin 2 (PS2) gene (on chromosome 1) are responsible for autosomal dominant early-onset Alzheimer's disease (EOAD). Missense mutations in these genes cause abnormal APP processing with subsequent overproduction of amyloidogenic and toxic A beta (42 peptide. A mutational analysis of APP, PS1, and PS2 genes can be used for both symptomatic and presymptomatic genetic testing and counselling in familial Alzheimer's disease (FAD). To contribute to our knowledge on genetic background of Alzheimer's disease in Poland, we screened APP mutations in a sample of familial EOAD cases from Poznan region. We did not find pathogenic mutations within exons 16 and 17 of the APP gene. Our study confirmed that APP gene mutations account only for a very small portion of FAD. PMID- 15119739 TI - Presenilin 1 mutations in Polish families with early-onset Alzheimer's disease. AB - Mutations in Presenilin 1 (PS1) and Presenilin 2 (PS2) genes account for up to 50% of familial early-onset Alzheimer's disease (EOAD). In order to assess the genetic contribution of the PS genes in a series of Polish patients, we performed a mutational analysis in 6 autosomal dominant (ADEOAD), 8 familial and 41 sporadic EOAD cases from Poznan region. Three missense mutations in the PS1 gene (Ala246Glu in exon 7, Pro267Leu in exon 8, and Leu424Arg in exon 12) were found in patients from families with ADEOAD. In addition, the Glu318Gly noncausative polymorphism in exon 9 was detected in two unrelated sporadic EOAD cases. The variation was also absent from other 53 patients and 48 controls. Therefore, we could not confirm the previous suggestion that the Glu318Gly substitution may be a risk factor for AD. PMID- 15119740 TI - Long-term effect of IFN-beta 1a therapy on CCL2 (MCP-1) chemokine in patients with multiple sclerosis. AB - Chemokines play an important role in pathogenesis of multiple sclerosis (MS), mediating migration of leukocytes into the central nervous system. CCL2 (MCP-1) chemokine is expressed in astrocytes in MS lesions. The aim of the study was to evaluate the effect of a two-year treatment with IFN-beta 1a on serum CCL2 level in MS patients. CCL2 concentration in sera of 18 relapsing-remitting MS (RR-MS) patients, and of 16 healthy controls was measured by ELISA. MS patients were treated with interferon-beta 1a (Avonex) in a dose of 30 microg i.m. once weekly. Significantly lower serum CCL2 level was found in MS patients in comparison with results of the control group. CCL2 concentration increased significantly after one year of therapy with IFN-beta, and remained high after the two-year treatment. The therapy of relapsing-remitting MS patients with interferon beta 1a is associated with a significant increase in CCL2 serum concentration. PMID- 15119741 TI - Association of influenza incidence with multiple sclerosis onset. AB - The aim of this study was to investigate the environmental factors influencing the tangible changes in the incidence of multiple sclerosis (MS) over a period of the past 30 years in the town Gniezno, Poland. We analysed many environmental factors to which the whole population was exposed in the respective period. The following factors were considered: viral infections (influenza, measles, varicella, rubella, mumps), atmospheric air pollution and climate conditions. A positive correlation has been found between the incidence of influenza and the incidence of MS in the same year (r = 0.37; p = 0.04) as well as with the MS incidence assessed after 5 years (r = 0.64; p = 0.0005). No significant correlation has been found between the incidence of MS and other investigated environmental factors. These results support the hypothesis that influenza infection could precipitate MS onset. PMID- 15119742 TI - Aneurysmal bone cyst of skull and vertebrae in children. Analysis of own material and review of the literature. AB - Aneurysmal bone cyst (ABC) is a benign, expansive, osteolytic lesion, consisting of blood-filled cysts, capable both of rapid enlargement and spontaneous resolution. Asymptomatic cases have been reported too. The aim of this paper was to analyse the outcomes of surgical treatment of ABC in children and a review of pertinent literature. We adopted the method of retrospective analysis of medical documentation of 10 patients with ABC. These were patients at the Department of Neurosurgery of the Children's Memorial Health Institute (Warsaw, Poland) from 1980 to 2002. There were 2 cases of cranial lesions and 8 cases of vertebral lesions. All the patients underwent surgical treatment only. Total lesionectomy was obtained in 7 cases, subtotal--in 3 cases. With the mean follow-up time of 5.1 years, good outcome (no neurological deficits) was noticed in 5 cases, moderate disability in the form of paraparesis--in 4 cases and full paraplegia- in 1 case. The following perioperative complications were noticed: transient paraplegia (1 case) and recurrence of ABC requiring reoperation (1 case). Among the 8 patients with vertebral lesions, progressive scoliosis requiring instrumental stabilization of the spine was noticed in 5 cases (4 vertebral body lesions and 1 laminar lesion). Primaiy infiltration of vertebral body by an ABC may lead to subsequent progressive scoliosis, which requires instrumental stabilization of spine. This demands careful planning and development of a comprehensive treatment program. ABC in children is a predominantly aggressive lesion, but even subtotal excision does not entrain a recurrence. Localization of lesion at the D3-5 levels is associated with an increased risk of postoperative neurological deterioration. Patients should be treated surgically before the development of severe deficits, which later may prove irreversible. PMID- 15119743 TI - Peripheral nerve tumours--diagnostic and therapeutical basics. AB - In this review of international literature, we have described recent opinions on diagnostics and therapy of peripheral nerve tumours. We have emphasised the use of differential diagnostics on certain stages of therapeutic procedures. The importance of proper surgical technique choice and its influence on final results have been especially underlined. Other important factors influencing final therapeutical results have also been considered and discussed. PMID- 15119744 TI - Hypertensive encephalopathy mimicking brainstem tumour in psychiatric patient. AB - Hypertensive encephalopathy is a syndrome consisting of headache, seizures, visual changes, and other neurologic disturbances in patients with elevated systemic blood pressure. Diagnosis based on clinical and radiological findings, which are not specific, may be difficult to establish. Furthermore, hypertensive encephalopathy may develop gradually even when blood pressure is lower than that of malignant hypertension. We present clinical, magnetic resonance imaging (MRI) and autopsy findings in a 43-year-old schizophrenic patient with unrecognised hypertensive encephalopathy, which was misinterpreted by MRI as a diffusely growing brain stem tumour. Increased blood pressure was recorded several times, but it was not properly controlled and treated either during his out-door psychiatric examinations or hospitalisation. At autopsy, generalised atherosclerosis, concentric hypertrophy of the left ventricle and arteriolonephrosclerosis were found in addition to microvascular fibrinoid necroses and thromboses in the brain and kidneys, which were almost certainly caused by arterial hypertension evolving from benign into malignant stage. We discuss the differential diagnosis and give a review of the literature. PMID- 15119745 TI - Multiple brain metastases from malignant peripheral nerve sheath tumour (MPNST). AB - Malignant peripheral nerve sheath tumours (MPNSTs) are rare soft tissue neoplasms arising from elements of the nerve sheath that often occur in the context of neurofibromatosis (NF) type 1. Their poor prognosis results from high local recurrence rate and distant dissemination. Nevertheless, the brain metastases are exceptional. We are presenting an unusual case of intrathoracic MPNST in a 33 year-old man with a five-year clinical course characterised by multiple times local recurrences of primary tumour and multiple remote metastases into the brain structures, thyroid and suprarenal gland. Moreover, the cerebellar metastasis regrew in spite of its total excision. Histologically, brain metastatic tumours were composed of spindle cells closely arranged in interlacing and woven fascicles. This highly cellular nerve tissue exhibited an advanced nuclear hyperchromasia and a high mitotic activity. The tumour exhibited rich delicate reticulin network. The schwannian nature of brain metastases has been confirmed by immunohistochemical findings showing S-100 protein and GFAP expression and ultrastructural evidences of the pericellular basal lamina. PMID- 15119746 TI - Very late relapse of medulloblastoma. AB - A case of 47-year-old woman with a local relapse of medulloblastoma 23 years after initial presentation is reported. At the age of 24, the patient underwent resection of medulloblastoma of the right cerebellar lobe, followed by the craniospinal orthovoltage irradiation (3600 R to the brain, and 3000 R to the spinal cord). At the 21st year of follow-up, a second cancer originating in the thyroid gland was diagnosed. Thyroidectomy followed by 131-iodotherapy for the papillary cancer was performed. Two years later she was operated for the recurrence of medulloblastoma at the former site. The patient was unfit for chemotherapy due to poor bone marrow reserve following the previous treatment. The reirradiation of the posterior cranial fossa was performed postoperatively. The patient was given 45 Gy in 25 fractions to the recurred tumour volume with 2 cm margin within 41 days. The treatment was performed by 6 MV photons with conformal technique and noncoplanar beams arrangement. The patient is disease free 15 months after relapse of medulloblastoma. The following problems are discussed: late relapse of medulloblastoma, secondary cancers after craniospinal irradiation, and retreatment of CNS tumours. PMID- 15119747 TI - Composite split cord malformation coexisting with spinal cord teratoma--case report and review of the literature. AB - The coincidence of split cord malformation (SCM) and intraspinal teratoma is an extremely uncommon condition. We report a case of a neurologically asymptomatic 13-year-old girl with a 5-year history of progressive scoliosis. The routinely performed MRI revealed SCM at levels L1-L2, tethering of the spinal cord at levels L4-L5 and the presence of an intraspinal tumour at the level of the observed SCM. Intraoperative investigation revealed composite character of SCM. The hemicords were separated by an osseo-cartilaginous septum and contained in a single dural tube. We claim that this is the second described case of a composite SCM and an intraspinal teratoma coexisting at the same level. This case supports the idea that scoliosis may be accompanied by congenital spinal cord anomalies like SCM and intradural tumours. MRI should be the basic diagnostic investigation in such cases. Scoliosis coexisting with SCM and an intradural tumour requires complex neurosurgical and orthopaedic treatment. PMID- 15119749 TI - [Educational program for asthmatic children and teenagers and their parents]. AB - BACKGROUND: Asthma is considered a health problem in Cuba. Its prevalence has increased and its morbidity and social-economic cost are important, although mortality and hospitalizations indicators have improved. Taking into consideration that the lack of information about this disease is considered a health hazard all over the world, an interactive educational program was prepared so that patients could learn how to handle their disease. MATERIAL AND METHODS: A two year, quasi-experimental intervention study was conducted with the participation of 50 children (29 boys and 21 girls from 8 to 15 years of age) and their parents. Healthcare, intake of reliever medications, pulmonary function, school attendance and the economic impact on family and on healthcare services were assessed. RESULTS: Educational program application conducted to a statistical significant lower consumption of reliever medication (p < 0.003), and emergency ward visits (p < 0.001). The patients improved their force expiratory volume in one second (p 0.003). Healthcare and family costs dropped to 56% and 18.13%, respectively. CONCLUSIONS: The intervention measures adopted had a positive effect and contributed to the reduction of morbidity and cost due to asthma. PMID- 15119750 TI - Circadian variation of cyclosporine A in renal transplanted patients. AB - BACKGROUND: A problem with the clinical use of cyclosporine A is the wide patient to-patient variability in blood levels. OBJECTIVE: To evaluate the circadian variation in patients with renal transplant on twice daily dosing in order to evaluate if patients could need a dosage adjustment in diurnal or nocturnal dose. MATERIAL AND METHODS: The circadian variation of oral cyclosporine pharmacokinetics at steady state was evaluated in 20 patients with renal transplant. Drug doses were administered daily at 8 am and at 8 pm and blood samples for cyclosporine levels measurement by radioimmunoassay were taken during the morning treatment and the evening treatment. RESULTS: Differences were found in the minimum steady state concentration between the morning and the evening administration, however no statistical differences were obtained in peak steady state concentration (Cmaxss), time to peak concentration (tmaxss), area under the curve at steady state (AUCss), mean residence time (MRT) and total clearance (Clt/F). CONCLUSIONS: Therapeutic drug monitoring of cyclosporine A is more important than the time administration. PMID- 15119751 TI - [Expression of leukocytic adhesion molecules in a patient with common variable immunodeficiency]. AB - BACKGROUND: Common variable immunodeficiency is one of the main antibodies' deficiency syndromes. OBJECTIVE: To present the immunological study of a 29-year old patient with common variable immunodeficiency who assisted to a check-up after being four years without treatment with gammaglobuline. MATERIAL AND METHODS: We studied a sample of peripheral blood and saliva of a patient with common variable immunodeficiency and that of a healthy patient (control). Assessment of immunoglobulin G, immunoglobulin A and immunoglobuin M was performed by a simple radial immunodiffusion test, and immunoglobulin E by immunoassay. Immunophenotypic study of leukocytic subpopulations was done by citometry by using the following panel of monoclonal antibodies: CD3 (Leu-4), CD4 (Leu-3a), CD8 (Leu-2a), CD19 (Leu-12), CD14 (Leu-M3), CD11a (LFA-I), CD49d (VLA 4), CD54 (ICAM-1), CD31 (PECAM). RESULTS: It was found a significant reduction in most of the serum and secretory immunoglobulins, levels of unusual expression of integrines CD11a and CD31 in lymphocytes T related to the low percentage of activated lymphocytes T/memory. PMID- 15119752 TI - [Assessment of the clinical efficacy and safety of epinastine plus pseudoephedrine vs loratadine plus pseudoephedrine in perennial allergic rhinitis]. AB - BACKGROUND: Several drugs for the treatment of perennial allergic rhinitis and its symptoms have been investigated since some years ago. These drugs are: steroidal-type immunoregulators, immunotherapy and antihistamines. Because of first generation of the last ones originated several side effects, pharmacological research was oriented to the search of formulas with the same of higher efficacy, but with fewer effects on central nervous system. OBJECTIVE: To compare the efficacy and safety of epinastine 10 mg plus pseudoephedrine 120 mg vs loratadine 5 mg plus pseudoephedrine 120 mg, twice a day, in the treatment of perennial allergic rhinitis. MATERIAL AND METHODS: A comparative, random, open, prospective, longitudinal and multicenter study was made in 62 patients with an average age of 26.17 +/- 9.75 years (13-56 years) with diagnosis of perennial allergic rhinitis, who attended to external consultation of the different departments of allergy of the participant institutions. All patients were submitted to: complete clinical history, physical exploration, measurement of vital signs, nasal exploration, qualification of allergic symptoms and record of daily symptoms. Moreover, washout during a week and pharmacological treatment was given during two weeks. Symptoms referred by patient and findings of physical exploration during the different visits, as well as rhinoscopic findings, were assessed by a scale of five parameters of rhinitis. Tolerance grade of drugs was assessed based on frequency and severity of side effects. RESULTS: According to the basal clinical assessment symptoms qualification of patients of groups of epinastine and loratadine was of 9.12 +/- 2.78 and 7.90 +/- 2.7, respectively. Seven side effects appeared: three cases of somnolence, one of sickness and one of anxiety in patients of group of loratadine and one case of somnolence and one of dysmenorrhea in the subjects of group of epinestine. Although it did not have statistically significant difference, it is clinically important for patients. Frequency of side effects was higher in the loratadine group. CONCLUSION: Both drugs are efficient for the treatment of perennial allergic rhinitis. PMID- 15119753 TI - [Sublingual vs subcutaneous immunotherapy]. AB - At the end of the 1980 decade, the first reports about the use of oral and sublingual routes of immunotherapy were published. This article reviews some reports comparing sublingual vs subcutaneous immunotherapy. PMID- 15119755 TI - Multimodal postoperative pain management. AB - PURPOSE: The mechanisms of persistent postoperative pain, rationale for multimodal pain therapy, and limitations of currently available analgesic agents and administration routes and techniques are described. SUMMARY: Persistent postoperative pain can cause long-term disability. It is the result of complex neurohormonal effects that can be prevented using preemptive analgesic therapy. Multimodal pain therapy can result in additive or synergistic analgesic effects and minimize adverse drug effects. Limitations of systemic opioid analgesics given by patient-controlled analgesia include adverse effects, a short duration of action, and nocturnal hypoxemia. Limitations of local anesthetics include unwanted motor blockade, which can interfere with postoperative mobilization and rehabilitation efforts. Inadvertent administration of analgesics into subarachnoid, subdural, or vascular spaces is a potential problem with the epidural route. Use of this route may be limited by the perioperative use of prophylactic anticoagulation. CONCLUSION: Research is needed to identify analgesic agents and administration techniques with greater efficacy and safety than those currently available. PMID- 15119754 TI - [Neurocysticercosis and asthma. Triggering or concomitant situation]. AB - The pleomorphism of neurocysticercosis makes its diagnosis impossible based on clinical data alone, thus, clinical data, epidemiologic backgrounds and neuroimaging are frequently used to do the diagnosis. On the other hand, the relationship between neurocysticercosis and allergic diseases is unclear. The present article discusses the coexistence of neurocysticercosis and asthma exacerbation in a patient in who two parenchymal cysts of the parasite were demonstrated. The enhancement of the allergic response and the parasite dye could be in relation with both allergic symptoms exacerbations and neurological symptoms appearance. PMID- 15119757 TI - Advances in postoperative pain management: the pharmacy perspective. AB - PURPOSE: The pharmacist's role in promoting proper use of analgesic drug therapies, by ensuring that Joint Commission on Accreditation of Healthcare Organizations (JCAHO) standards for pain management are met, evaluating new analgesic options for formulary addition, and conducting medication-use evaluations, outcomes research, and pharmacoeconomic analyses, is discussed. SUMMARY: Pharmacists can be instrumental in meeting JCAHO requirements for educating clinical staff and patients about pain management practices and monitoring compliance with JCAHO pain management standards. They also can thoroughly evaluate new analgesic drug therapies and delivery systems for formulary addition by reviewing published literature and manufacturers' data and conducting medication-use evaluations, outcomes research, and pharmacoeconomic analyses. Cost-effectiveness analysis is the most common among the four types of pharmacoeconomic analyses, which include cost-minimization analysis, cost-benefit analysis, and cost-utility analysis. A ten-step process is used in conducting pharmacoeconomic analyses to increase the validity of the results. CONCLUSION: Implementation of new analgesic drug delivery technologies, such as an elastomeric infusion pump (a device for continuous wound infiltration with a local anesthetic), a needle-free transdermal patient-controlled analgesia system, and a new sustained-release, liposome-encapsulated form of morphine for epidural injection, offers new postoperative pain management alternatives for patients and clinical staff and involves unique considerations for pharmacists. PMID- 15119756 TI - Emerging techniques in the treatment of postoperative pain. AB - PURPOSE: Causes of inadequate postoperative pain control, challenges with currently available analgesic therapies, characteristics of optimal postoperative analgesic therapy, a liposome-encapsulated, sustained-release dosage form of morphine, and other investigational analgesic therapies are described. SUMMARY: Technical difficulties in analgesic drug administration, inappropriate prescribing, unfounded patient fears or expectations, and the lack of availability of parenteral dosage forms of certain analgesic agents can potentially contribute to inadequate postoperative pain control. The use of epidural catheters for analgesic therapy is limited by the use of prophylactic anticoagulation, the tendency for catheters to migrate and fail, and the need for nursing staff time to monitor the catheter site and infusion pump. Continuous peripheral blocks are not always effective, the devices used are bulky, and the technology is prone to failure. Multimodal therapies are labor-intensive, and some components must be given orally because they are not available in parenteral dosage forms. Optimal postoperative analgesic drug therapy would therefore be noninvasive, easy to administer, require no cumbersome equipment, cause no or minimal adverse effects, and be safe to use in patients receiving prophylactic anticoagulation. A new morphine sulfate sustained-release liposome injection has several of these characteristics, including ease of administration and compatibility with anticoagulation. Single-dose epidural administration of morphine sulfate sustained-release liposome injection in patients undergoing hip arthroplasty provided better pain control than i.v. opioid analgesics given by PCA, with similar adverse effects. CONCLUSION: Opioid analgesics remain the mainstay of postoperative analgesic therapy, but the development of infusion pumps with improved reliability; needle-free technologies, parenteral dosage forms of acetaminophen, selective cyclooxygenase-2 inhibitors, and other non opioid therapies; and new opioid analgesic agents that provide analgesia without the adverse effects of currently available opioid analgesics may substantially improve postoperative analgesia and health-related quality of life in the postoperative pain patient in the future. PMID- 15119758 TI - Zoonotic viruses of wildlife: hither from yon. AB - The emergence of zoonotic viruses maintained by wildlife reservoir hosts is poorly understood. Recent discoveries of Hendra (HENV) and Nipah (NIPV) viruses in Australasia and the emergence of epidemic West Nile virus (WNV) in the United States have added urgency to the study of cross-species transmission. The processes by which zoonotic viruses are transmitted and infect other species are examined as four transitions. Two of these, inter-species contact and cross species virus transmission (spillover), are essential and sufficient to cause epidemic emergence. Sustained transmission and virus adaptation within the spillover host are transitions not required for virus emergence, but determine the magnitude and scope of subsequent disease outbreaks. Ecologic, anthropogenic, and evolutionary factors modify the probability that viruses complete or move through transitions. As surveillance for wildlife diseases is rare and often outbreak-driven, targeted studies are required to elucidate the means by which important zoonotic viruses are maintained and spillover occurs. PMID- 15119759 TI - The role of surveillance in polio eradication and identification of emerging viral encephalitis. AB - In 1988, when the World Health Assembly resolved to eradicate paralytic poliomyelitis, polio was endemic in 125 countries on 5 continents with an estimated 350,000 cases annually. By 2002, the number of countries was reduced to 7 and the number of cases by greater than 99%. Instrumental in this extraordinary progress is the timely detection and investigation of all cases of acute flaccid paralysis and the examination of stool samples in an accredited WHO Global Network laboratory. The Network consists of 124 National Poliovirus Laboratories, 15 Regional Reference Laboratories, and 7 Global Specialized Laboratories. Network Laboratories are held accountable to rigid performance standards for quality and timeliness in testing more than 60,000 stool samples annually. The Network is a hierarchical system in which polioviruses are isolated and identified in National Laboratories, differentiated as to wild or vaccine origin in Regional Reference Laboratories, and sequenced in Specialized Laboratories. Findings are promptly relayed to Regional and national program managers for immunization strategic planning, monitoring surveillance quality, and assessing eradication progress. Lessons from the Polio Laboratory Network demonstrate the value of a central coordinating body, an effective communication infrastructure, and full partnerships with peer epidemiology and medical sectors. Post eradication polio surveillance will continue for many years. The Network legacy for other public health initiatives is access to an existing laboratory infrastructure and human resources with the proven ability to achieve technology transfer and quality laboratory performance even in resource-poor countries. PMID- 15119760 TI - Emergence and virulence of encephalitogenic arboviruses. AB - Each arbovirus that causes encephalitis is geographically restricted by the availability of appropriate vectors and reservoir hosts. These viruses evolve regionally by recombination, reassortment and point mutation and can "emerge" as causes of human encephalitis through extension to new geographic regions or by selection of more virulent or more efficiently transmitted virus variants. The properties of arboviruses that result in encephalitis involve efficient replication in peripheral tissues after initiation of infection, production of a viremia, entry into the central nervous system and efficient replication in neurons with spread to additional populations of neurons. Many of these steps are determined by properties of the envelope glycoproteins responsible for cellular attachment, but changes in noncoding regions of the genome, as well as in other structural and nonstructural proteins, also contribute to neurovirulence. PMID- 15119761 TI - Molecular determinants of virulence of West Nile virus in North America. AB - West Nile virus (WNV) is a mosquito-borne flavivirus that until very recently had not been found in the Americas. In 1999, there was an outbreak of West Nile encephalitis in New York and surrounding areas, involving 62 human cases, including 7 fatalities. The virus has subsequently become established in the United States of America (U.S.) with 4156 human cases, including 284 deaths, in 2002. The WNV strains found in the U.S. are members of "lineage I", a genetic grouping that includes viruses from Europe, Asia and Africa. Molecular epidemiologic studies indicate that two genetic variants of WNV emerged in 2002. The major genetic variant is found in most parts of the U.S., while the minor genetic variant has been identified only on the southeast coast of Texas. Investigation of WNV in mouse and hamster models demonstrated that strains from the U.S. are highly neurovirulent and neuroinvasive in these laboratory rodents. Other strains, such as Ethiopia 76a from lineage I, are not neuroinvasive and represent important viruses which can be used to elucidate the molecular basis of virulence and attenuation of WNV. To identify putative molecular determinants of virulence and attenuation, we have undertaken comparative nucleotide sequencing of Ethiopia 76a and strains from the U.S. The results show that the two viruses differ by 5 amino acids in the envelope (E) protein, including loss of the glycosylation site. Comparison of our panel of 27 WNV strains suggests that E protein glycosylation is a major determinant of the mouse neuroinvasive phenotype. PMID- 15119762 TI - Genetic determinants of Venezuelan equine encephalitis emergence. AB - Following a period of inactivity from 1973-1991, Venezuelan equine encephalitis (VEE) reemerged during the past decade in South America and Mexico. Experimental studies of VEE virus (VEEV) infection of horses with virus strains isolated during these outbreaks have revealed considerable variation in the ability of equine-virulent, epizootic strains to exploit horses as efficient amplification hosts. Subtype IC strains from recent outbreaks in Venezuela and Colombia amplify efficiently in equines, with a correlation between maximum viremia titers and the extent of the outbreak from which the virus strain was isolated. Studies of enzootic VEEV strains that are believed to represent progenitors of the epizootic subtypes support the hypothesis that adaptation to efficient replication in equines is a major determinant of emergence and the ability of VEEV to spread geographically. Correlations between the ability of enzootic and epizootic VEEV strains to infect abundant, equiphilic mosquitoes, and the location and extent of these outbreaks, also suggest that specific adaptation to Ochlerotatus taeniorhynchus mosquitoes is a determinant of some but not all emergence events. Genetic studies imply that mutations in the E2 envelope glycoprotein gene are major determinants of adaptation to both equines and mosquito vectors. PMID- 15119763 TI - Evolution and dispersal of encephalitic flaviviruses. AB - There are two major groups of encephalitic flaviviruses, those that infect and are transmitted by ticks, particularly Ixodes spp. and those that infect and are transmitted by mosquitoes, particularly Culex spp. The tick-borne encephalitic flaviviruses exhibit evolutionary characteristics that are largely determined by the protracted life cycle of the tick, its habitat and the prevailing climatic conditions. These viruses appear to have evolved gradually from non-encephalitic viruses that radiated eastwards and north eastwards out of Africa into Asia and the southern islands, then northwards to far east Asia and finally westwards across Eurasia to western Europe, during the past two to four thousand years. Only one of these recognized species has found its way to North America viz. Powassan virus. In contrast, the evolution of the recognized mosquito-borne encephalitic flaviviruses reflects the wide range of mosquito species that they infect. They emerged out of Africa relatively recently and at roughly the same time, i.e., probably during the past few centuries. Although many of these mosquito-borne viruses are geographically widely dispersed, with the exception of West Nile virus, they are found either in the Old World or the New World, never in both, and we are now beginning to understand the reasons. Phylogenetic trees will be used here to describe the evolution, epidemiology and dispersal characteristics of these viruses, taking into account the importance of virus persistence and recombination. PMID- 15119765 TI - Emerging encephalitogenic viruses: lyssaviruses and henipaviruses transmitted by frugivorous bats. AB - Three newly recognized encephalitogenic zoonotic viruses spread from fruit bats of the genus Pteropus (order Chiroptera, suborder Megachiroptera) have been recognised over the past decade. These are: Hendra virus, formerly named equine morbillivirus, which was responsible for an outbreak of disease in horses and humans in Brisbane, Australia, in 1994; Australian bat lyssavirus, the cause of a severe acute encephalitis, in 1996; and Nipah virus, the cause of a major outbreak of encephalitis and pulmonary disease in domestic pigs and people in peninsula Malaysia in 1999. Hendra and Nipah viruses have been shown to be the first two members of a new genus, Henipavirus, in the family Paramyxoviridae, subfamily Paramyxovirinae, whereas Australian bat lyssavirus is closely related antigenically to classical rabies virus in the genus Lyssavirus, family Rhabdoviridae, although it can be distinguished on genetic grounds. Hendra and Nipah viruses have neurological and pneumonic tropisms. The first humans and equids with Hendra virus infections died from acute respiratory disease, whereas the second human patient died from an encephalitis. With Nipah virus, the predominant clinical syndrome in humans was encephalitic rather than respiratory, whereas in pigs, the infection was characterised by acute fever with respiratory involvement with or without neurological signs. Two human infections with Australian bat lyssavirus have been reported, the clinical signs of which were consistent with classical rabies infection and included a diffuse, non suppurative encephalitis. Many important questions remain to be answered regarding modes of transmission, pathogenesis, and geographic range of these viruses. PMID- 15119764 TI - West Nile virus and other zoonotic viruses in Russia: examples of emerging reemerging situations. AB - Studies of the interactions of vertebrates, viruses and arthropod vectors of these viruses were monitored in terms of different ecological groups of viruses transmitted by mosquitoes and ticks in Northern Eurasia in an area encompassing more than 15 million km2. About 90 viruses were isolated, including 24 new to science. Newly recognized infections of vertebrates, including humans, were described. Many unusual epidemic situations were analysed. Permanent efforts were established to prevent bioterrorist activities and their consequences. Extensive epidemic outbreaks of West Nile fever (WNF; i.e., fever caused by West Nile virus) and Crimean-Congo hemorrhagic fever (CCHF) with unusual high mortality appeared in the last four years in southern Russia. We determined infection rates in humans, domestic and wild animals, mosquitoes and ticks from natural and synanthropic biocenoses [Editorial note: "synanthropic" means, roughly, all species living with (c.f. lice, fleas) or near people, such as in houses (c.f. house mice), parks (c.f. Rattus spp.), and the like, rather like "peridomestic", but not strictly so; "biocenosis" is the biome, the "totality of living populations in a particular habitat, which itself is only a part of the ecosystem".]. CCHF virus strains were phylogenetically similar to strains isolated in this area 35 years ago but different from Central-South-Asian and African strains. Before the outset of the current emergence of epidemic WNF, three genetic variants of this virus had been isolated in USSR, two African and one Indian. Phylogenetic analysis of complete genome sequences of epidemic strains demonstrated considerable similarity to strains from USA and Israel and differences from strains isolated in the same USSR areas 20-30 years before. In addition to strains of genotype 1, we isolated strains of second and third lineages and a strain of a fourth genetic variant. Nucleotide differences of these strains from all three genotypes was about 30%. The emerging WNF situation in Russia for the last 4 years probably has been the result of not only natural and social factors, but also to introduction of more virulent strains or by evolution of the virus. PMID- 15119766 TI - Novel viral encephalitides associated with bats (Chiroptera)--host management strategies. AB - Several novel viruses recently described in bats of the genus Pteropus (sub-order Megachiroptera) in Australia and southeast Asia cause encephalitic disease in animals and humans. These viruses include Hendra virus and Nipah virus (genus Henipavirus, family Paramyxoviridae) and Australian bat lyssavirus (ABLV; genus Lyssavirus, family Rhabdoviridae). Broadly, strategies for disease prevention and control in the spillover host are directed at minimising direct or indirect contact with the natural host, improving farm-gate and on-farm biosecurity, and better disease recognition and diagnosis. Additional strategies for ABLV include the use of rabies vaccine for effective pre- and post-exposure prophylaxis in humans. Effective management strategies in the natural host are predicated on an understanding of the ecology of the disease in the natural host, and the identification and avoidance of factors putatively associated with emergence, such as habitat loss, land use change and demographic shifts. A possible future management strategy for ABLV in reservoir populations is immunisation using bait or plant-derived vaccination. PMID- 15119768 TI - The entry machinery of flaviviruses. AB - We have been using the flavivirus tick-borne encephalitis virus (TBEV) as a model system for investigating the molecular mechanisms underlying the membrane fusion process mediated by a class II viral fusion protein, the flavivirus envelope protein E. In the mature virion this protein exists as a metastable dimer that dissociates at the acidic pH in endosomes and is converted into a more stable trimeric conformation. The dimer dissociation step liberates an internal fusion peptide that interacts with the target endosomal membrane, and then further conformational changes are believed to drive membrane fusion. Although flavivirus fusion appears to be a more facile and efficient process than that of alphaviruses, which also possess a class II viral fusion protein, the fusion mechanism in both viral systems involves structurally related interactions with lipids, specifically the 3beta-hydroxyl group at C3 of cholesterol. The class II viral fusion machineries are structurally different from those involving class I viral fusion proteins, such as those found in orthomyxoviruses, paramyxoviruses, retroviruses, and filoviruses, but have certain similarities in common with bacterial pore-forming proteins. PMID- 15119767 TI - Henipaviruses: recent observations on regulation of transcription and the nature of the cell receptor. AB - Hendra virus (HENV) and Nipah virus (NIPV) are classified in the new genus Henipavirus, within the subfamily Paramyxovirinae, family Paramyxoviridae. The genetic and biological characteristics that differentiate henipaviruses from other members of the subfamily are summarized. Although they do not display neuraminidase and hemagglutination activities and in that regard resemble viruses in the genus Morbillivirus, several recent observations highlight similarities between henipaviruses and respiroviruses (genus Respirovirus) in structure and replication strategy. First, three-dimensional modeling studies suggest that the external globular head domain of the HENV G protein resembles that of respiroviruses rather than morbilliviruses. Second, the pattern of transcriptional attenuation in HENV-infected cells resembles that observed with Sendai virus, a respirovirus, and differs from that found in cells infected with measles virus, a morbillivirus. Henipaviruses have a broad host range in vitro and in vivo, indicating wide distribution of cellular receptor molecules. The extensive host range has been confirmed in a quantitative in vitro cell-fusion assay using recombinant vaccinia viruses expressing the attachment and fusion proteins of HENV and NIPV. Cell lines of diverse origin and which are permissive in the in vitro cell fusion assay have been identified and the pattern of relative susceptibilities is the same for both HENV and NIPV, implying that both viruses use the same cell receptor. Protease treatment of permissive cells destroys their ability to fuse with cells expressing viral envelope glycoproteins. Virus overlay protein binding assay (VOPBA) and radio-immune precipitation assays confirm that both HENV and NIPV bind to membrane proteins in the 35-50 kD range. Treatment of cell membrane proteins with N-glycosidase eliminates HeV binding activity in VOPBA whereas treatment with neuraminidase has no effect on binding. Thus preliminary evidence suggests that NIPV and HENV bind to the same glycoprotein receptor via a non-sialic acid-dependant mechanism. PMID- 15119769 TI - Persistent infection and suppression of host response by alphaviruses. AB - Alphaviruses cause chronic noncytopathic infection in mosquito cells and develop a highly cytopathic infection in a wide variety of cells of vertebrate origin. Upon infection, alphaviruses modify cellular processes to meet the virus needs for propagation. Downregulation of translation and transcription caused by viral infection appears to reduce interferon (IFN) and cytokine gene expression and allows more efficient dissemination of infection. Alphaviruses with mutations in nonstructural protein nsP2 can become less cytopathic and capable of persisting in some vertebrate cell lines for a number of passages. nsP2 likely functions as an important regulator of virus-host cell interactions and plays a significant role in suppressing the antiviral response. Mammalian cells having no defects in type I IFN system react to replication of the nsP2 viral mutants by more efficient activation of IFN and IFN-dependent genes and are capable of eliminating established alphavirus infection. Blocking of IFN-alpha/beta signaling makes mouse fibroblasts unable to stop replication of Sindbis virus (SINV) with mutated nsP2 and leads to persistent infection. Downregulation of transcription and translation during alphavirus infection are quite independent events, and both probably are involved in inhibition of the antiviral response. PMID- 15119770 TI - Subversive neuroinvasive strategy of rabies virus. AB - Rabies virus (RABV) is a pathogen well-adapted to the nervous system, where it infects the neurons. RABV is transmitted by the bite of an infected animal. It enters the nervous system via a motor neuron through the neuromuscular junction, or via a sensory nerve through nerve spindles. It then travels from one neuron to the next, along the spinal cord to the brain and the salivary glands. The virions are then excreted in the saliva of the animal and can be transmitted to another host by bite. Thus, preservation of the neuronal network integrity is crucial for the virus to be transmitted. Successful invasion of the nervous system by RABV seems to be the result of a subversive strategy based on the survival of infected neurons including protection against virus-mediated apoptosis and destruction of T cells that invade the CNS in response to infection. PMID- 15119771 TI - Neurovirulence and host factors in flavivirus encephalitis--evidence from clinical epidemiology. AB - Japanese encephalitis virus (JEV) and West Nile virus (WNV) provide some of the most important examples of emerging zoonotic viral encephalitides. For these flaviviruses, only a small proportion of those infected develop clinical features, and these may range from a non-specific flu-like illness to a severe fatal meningoencephalitis, often with Parkinsonian features, or a poliomyelitis like flaccid paralysis. The factors governing the clinical presentations, and outcome of flavivirus infections are poorly understood, but studies have looked at viral virulence determinants and the host immune response. Previous studies on JEV have suggested that the distribution of the four genotypes across Asia may relate to the differing clinical epidemiology (epidemic disease in the north, endemic disease in the south). However, new data based on the complete nucleotide sequence of a virus representing one of the oldest lineages, and phylogenetic analyses of all JEV strains for which genetic data are available, suggest that the distribution is best explained in terms of the virus' origin in the Indonesia Malaysia region (where all genotypes have been found), and the spread of the more recent genotypes to new geographical areas. Clinical studies have shown that innate immunity, as manifested by interferon alpha levels, is important in JEV and other flaviviruses, but treatment with interferon alpha did not improve the outcome. A failure of the humoral immune response, is associated with death from encephalitis caused by JEV and WNV. Cellular immunity has been less well characterized, but CD8+ and CD4+ T cells are thought to be important. PMID- 15119773 TI - Semliki forest virus infection of laboratory mice: a model to study the pathogenesis of viral encephalitis. AB - Semliki Forest virus (SFV) infection of the laboratory mouse provides an experimental system to study the pathogenesis of viral encephalitis. Following extraneural inoculation the virus is efficiently neuroinvasive and crosses the blood-brain barrier to initiate perivascular foci of infection in neurons and oligodendrocytes. The outcome of infection ranges from clinically unapparent mild encephalitis to fatal panencephalitis. SFV infections of the developing nervous system are always highly destructive and are generally fatal. In contrast, SFV infections of the mature nervous system can result in persistent infection with no apparent cell loss. This dramatic difference is attributable to developmental changes in the interactions between virus and CNS cells. Antibody responses clear the systemic infection and control the CNS infection. CD8+ T-cells are required to generate the lesions of inflammatory demyelination which can be a feature of the neuropathology. This article reviews the pathogenesis of SFV encephalitis, describing the neuropathology and the mechanisms which underlie it and which may be fundamental to many viral encephalitides. PMID- 15119772 TI - Regulation of apoptosis by viruses that infect insects. AB - Orthobunyviruses and alphaviruses cause encephalitis, neuronal apoptosis and mortality in mammals, but fail to kill the mosquitoes that transmit these viruses. Therefore, host cell factors, as well as viral factors, regulate the outcome of infection. Drosophila Reaper is a pro-death factor in the Drosophila nervous system during development but homologues were not previously known to exist outside flies. Recent discovery of a Reaper protein encoded by orthobunyaviruses provides interesting insight into the mechanisms by which viruses modulate the death pathway in both vertebrate and invertebrate hosts. PMID- 15119774 TI - A novel principle of attenuation for the development of new generation live flavivirus vaccines. AB - The genus Flavivirus includes a number of important human pathogens that impose major health problems in large regions of the world. The emergence of flaviviruses in new geographic regions (e.g., West Nile virus in North America) and rapid socioeconomic changed in many developing countries where flaviviruses such as dengue virus and Japanese encephalitis virus and endemic demand the development of new vaccines against these diseases. Using tick-borne encephalitis virus as a model we have established a new method to generate attenuated flavivirus strains that may be useful for generating cost-effective and safe live vaccines. This method relies on the specific introduction of deletions into one of the structural proteins, the capsid protein C. These deletions remove parts or all of an internal stretch of hydrophobic amino acid residues that probably is involved in virion assembly. We observed that remarkably long deletions were tolerated, yielding viable viral mutants that were highly attenuated in the mouse model but efficiently induced protective immunity. Biochemical analyses suggested that attenuation was caused by an assembly defect of infectious virions but the mutants produced ample amounts of non-infections subviral particles. The generation of viable mutants with deletions longer that 16 amino acid residues depended on additional, spontaneously emerging mutations within protein C that increased the hydrophobicity of the mutant protein. Although the second-site mutations increased infectivity, they did not restore neuroinvasiveness. Mouse experiments demonstrated excellent safety and immunogenicity profiles for these mutants. PMID- 15119775 TI - Tick-borne encephalitis and the impact of vaccination. AB - TBE virus is endemic in many parts of Europe and Northern Asia, and in these regions it causes more than 10,000 severe cases of central nervous system disease in humans each year. The virus is primarily transmitted to humans when infected ticks take a blood meal, but infections due to the consumption of unpasteurized milk, primarily from goats, occur in certain regions. Based on genetic analyses, three closely related subtypes can be distinguished and are designated European, Siberian, and Far Eastern subtype according to their primary geographic distribution. Consistent with their close antigenic relationships, immunization studies in animals have revealed a high degree of cross-protection between virus strains belonging to different subtypes. The commercially available vaccines in Europe consist of highly purified inactivated whole TBE virus. Austria is the country with the highest coverage of TBE vaccination (86% of the total population) and this has led to a dramatic reduction in the annual number of clinical cases and proves under field conditions that vaccination is an effective means for the prophylaxis of TBE. PMID- 15119776 TI - Future vaccines against emerging encephalitides. AB - The vaccine strategies available for control of emerging encephalitides range in a continuum from traditional approaches to those utilizing new technologies. In this report, we explore the use of live attenuated vaccines where the attenuating mutations have been selected in a rational way to improve attenuation without sacrificing effectiveness. A strategy for paired lethal and resuscitating mutations is presented that will greatly reduce the possibility of reversion to virulence. Finally, we describe an example of a vaccine vector system that could be rapidly adapted for use against these virus diseases as they emerge. PMID- 15119777 TI - Recombinant canarypoxvirus vaccine carrying the prM/E genes of West Nile virus protects horses against a West Nile virus-mosquito challenge. AB - An ALVAC (canarypoxvirus)-based recombinant (vCP2017) expressing the prM and E genes derived from a 1999 New York isolate of West Nile virus (WNV) was constructed and assessed for its protective efficacy in horses in two different experiments. In the first trial, a dose titration study was conducted to evaluate both serum neutralising antibody responses to WNV and duration of immunity. In the second trial the onset of protection was determined. Twenty-eight adult horses received two doses of vCP2017 administered intramuscularly at 5-week intervals and sixteen horses comprised age-matched non-vaccinated controls. Individual sera were taken periodically and tested for neutralising antibodies against WNV. Horses were challenged by allowing WNV-infected Aedes albopictus mosquitoes to feed on them two weeks (second trial) or one year (first trial) after the second vaccination. After challenge, horses were monitored for clinical signs of disease, and blood samples were collected for detection of WNV viremia and antibody. In both trials, all vaccinated horses developed neutralising antibodies against WNV. None of the vaccinated or control horses developed clinical signs of WNV disease upon challenge. None of the nine horses challenged 2 weeks after primary vaccination and only one of the ten vaccinated horses challenged 1 year after vaccination developed detectable viremia after challenge, whereas more than 80% of the controls became infected. Results from these studies demonstrated that a primary course of two doses of vCP2017 provides both antibody response and an early immunity in horses against WNV viremia. PMID- 15119778 TI - Diagnosis of zoonotic viral encephalitis. AB - Human infections by zoonotic encephalitis viruses are usually asymptomatic or symptoms are not specific to these viruses. Some of them have high mortality and morbidity rates and most often no specific treatment exist. This emphasizes the need for a precise identification of arboviruses in clinical specimens from humans and animals. Because these diseases are frequent in developing countries and tend to emerge or re-emerge in others, diagnostic tools must detect the broadest possible range of viruses with a high sensitivity and this is a key factor for surveillance, control of transmission and prevention through vaccination. In countries with limited diagnostic infrastructures, low-cost and easy-to-use tests are required. The diagnosis of arboviral encephalitis has been significantly improved in the recent years. Sensitive ELISA assay to detect antibodies against many arboviruses in serum or CSF are commercially available and can be used to detect early infections. Immunochromatographic rapid tests for the detection of specific IgM that could be used on fingertip blood would be valuable tools in developing countries. A limitation of these serologic assays is their lack of specificity as many arboviruses are antigenically related. Virus isolation or molecular assays from different human or animal tissues are also important diagnostic tools. Molecular assays have been extensively described in the recent years. They are very sensitive and have the advantage over cell culture that specimen transportation is less critical. Real-time detection has even improved sensitivity and reduced time-to-result. Although the utility of molecular assays for the detection of arboviruses in mosquito pools has been demonstrated, an extensive validation of their pertinence in clinical settings is still required. The use of DNA-microarrays may further extend the range of viruses that can be detected in a single test and allow isolates typing for epidemiology purposes. PMID- 15119779 TI - [The balanced force and the GT-rotary technique in comparison with the non instrumental technique (NIT)]. AB - The aim of the study was to compare the cleansing effect of the latest modification of the non-instrumentation technique (NIT) to that of conventional instrumentation. The root curvature in 100 vital human molars was determined by a standardized X-ray procedure and the teeth were assigned to five groups with 20 teeth each with an equal distribution of the root curvature. The preparation methods were the Balanced Force technique and the GT Rotary technique. Each root was irrigated with 40 ml of 3% sodium hypochlorite. The other groups were irrigated by NIT during 2.5, 5 or 10 minutes, respectively. The remaining pulpal tissue was stained and the root canals were exposed longitudinally. The teeth were then evaluated using a microscope and an image analysis-system. The residual organic debris in the apical, middle and coronal sections of the root canals were assessed as a percentage of the corresponding total examined length. The cleansing effect of the NIT in the coronal and middle parts of the canal used for 5 and 10 minutes was significantly better (p < 0.05) compared to using the device for 2.5 minutes. The cleansing effect of the NIT in the coronal and middle parts of the canal used for 5 and 10 minutes was also significantly better (p < 0.05) compared to using the GT Rotary or Balanced Force techniques. Apically, the cleansing effect of the NIT used for 5 and 10 minutes and the GT Rotary technique was significantly better (p < 0.05) compared to using the Balanced Force technique or the NIT for 2.5 minutes. It was concluded that the cleansing effect of the latest modification of the Non-instrumentation Technology (NIT) was equivalent to or better than that of conventional instrumentation requiring significantly less time. PMID- 15119780 TI - [Intraoral piezo-surgery]. AB - The Piezo "effect"describes a physical interaction, which is used in different fields of clinical medicine. An ultrasound technology results in the power of a surgical cut technique, which is able to distinguish between hard and soft tissue. With the presented instruments it is possible to treat bone mechanically in a piezo-surgical manner without hurting the surrounding soft tissues when touching them. Four examples: Tooth extraction before implantation under minimal grinding of bone, Gaining bone for peri-implant transplantation, Preparation of Sinus Lift, Exposing the inferior alveolar nerve demonstrate the use of Piezo Surgery in the intraoral area. This method shows a remarkable addition to the intraoral operation techniques. PMID- 15119781 TI - [Amine fluoride does not cause disintegration of glass ceramics]. AB - Topical application of fluoride has been proven to be an effective method for the prevention of dental caries. Some authors have, however, pointed at the etching potential of topically applied fluoride and argued that some products might etch and roughen the surface of ceramic restorations. The aim of the present study was to assess whether the topical application of an acidulated fluoride gelee in vivo leads to a damage of glass ceramic. It was shown that individual brushing with an acidulated fluoride gelee according to the manufacturers prescription is not harmful to ceramic restoration material. PMID- 15119782 TI - Arterial supply of the pineal gland of Akkaraman sheep. AB - Arterial vascularisation of the ovine pineal gland was investigated by latex injection preparations of the common carotid artery in 20 adult Akkaraman sheep brains under stereo light microscope. It was observed that the arterial supply comes exclusively from branches of the caudal cerebral artery. The pineal gland was found to contain a rich vascular network. This network also received a few branches from the caudal choroid rami. PMID- 15119784 TI - The incidence of mastitis treated with antibiotics in large-scale Hungarian Holstein-Friesian dairy farms. AB - Treated mastitis episodes at large Hungarian dairy farms were studied to determine the distribution of mastitis treated with antibiotics among quarters of the udder. Data were detailed records of all mastitis episodes that occurred during 1976 lactations in Farm A infected with Staphylococcus aureus (from May 1995 through July 1998) and 808 lactations in Farm B free from S. aureus (from January 1999 through March 2001). The distribution of treated quarters was compared with mathematical expectations based upon a random distribution in the case of Farm A. Results on mastitis incidences for different lactation stage groups showed an increasing incidence within subsequent lactation stage groups in Farm A. In contrast, in Farm B the mastitis incidence for lactation stage group between 35 and 100 days was the highest, but beyond 100 days the incidence decreased and reached the lowest value. Results gave strong evidence that the four quarters within the udder are not distributed randomly with respect to naturally occurring episodes of treated mastitis. More episodes than expected occurred in which only one or all four quarters were treated. Fewer episodes than expected with two or three treated quarters were observed. In both farms, the mastitis rate for rear quarters was higher than for front quarters, and the incidence of right quarter mastitis episodes was higher than that of left quarter mastitis episodes. PMID- 15119783 TI - Staphylococci isolated from animals and food with phenotypically reduced susceptibility to beta-lactamase-resistant beta-lactam antibiotics. AB - The antibiotic resistance pattern of 1921 Staphylococcus strains isolated from animals and food within the last two years were examined using diffusion tests. Among them there were only 35 strains of S. aureus having an inhibition zone diameter of 15 mm or less, and 4 strains of coagulase-negative staphylococci (CNS) having a zone diameter of 18 mm or less to 1-microg oxacillin disk. These 39 strains were examined also by E-test to oxacillin and for the detection of the mecA gene by PCR in order to determine whether they might be real methicillin resistant staphylococci. Among the 39 strains there were only two that were susceptible to penicillin by disk diffusion method; however, further examination by the penicillinase test showed that they produced beta-lactamase. While 19 (15 S. aureus, 4 CNS) strains were resistant and 7 strains were intermediate to oxacillin in disk diffusion test, the E-test gave 8 resistant and 5 intermediate results. Six out of the 8 oxacillin-resistant strains examined by disk diffusion and E-test harboured the mecA gene. Thus only 6 out of the examined 1921 strains proved to be mecA positive. These methicillin-resistant, mecA-positive strains (5 of the S. aureus strains and 1 of the S. epidermidis) originated from two dairy herds. The results prove that methicillin-resistant S. aureus (MRSA) strains in animals are really rare in Hungary. Eighteen strains were chosen and screened for minimal inhibitory concentration (MIC) of oxacillin with or without clavulanic acid or sulbactam, and three of them produced methicillinase enzyme. PMID- 15119785 TI - Effects of periodontal dressings on fibroblasts and gingival wound healing in dogs. AB - In the present study the effects of different commercially available periodontal dressings (Peripac, Barricaid, Fittydent, Reso-Pack and Myzotect-tincture) on fibroblast (V-79-379A) proliferation and survival were tested in vitro. Barricaid, Fittydent and Reso-Pack periodontal dressings have only small inhibitory effects on cell proliferation (83.3 +/- 9%, 71.6 +/- 8.7% and 87.3 +/- 4.5% of control after 48 h, respectively) in comparison with the great inhibitory effect of Myzotect-tincture (2.9 +/- 0.1%) and Peripac (33.7 +/- 11.4%) (p < 0.001). Barricaid was the only dressing where 41% of cells survived after exposure, while the other four dressings killed all the cells in 6 days. In addition, the healing of artificially created gingival wounds covered by Barricaid and Reso-Pack was followed for 7 days in 12 Beagle dogs. Histological evaluation of gingival tissue demonstrated that wounds covered by Reso-Pack showed the best epithelisation and vascularity and the least inflammatory reaction in first 4 days. Later the observed parameters were similar with those of wounds covered by Barricaid or without pack. The present results indicate that Peripac periodontal dressing and Myzotect-tincture showed the highest cytotoxicity to fibroblasts in vitro. From the histological observations in Beagle dogs Reso-Pack has been found to be the most suitable dressing, followed by Barricaid. PMID- 15119786 TI - Successful surgical removal of fibroma from the uterus of a cow: a case report. AB - A Holstein-Friesian cow aged 6 years aborted twice at 3-4 months of gestation. On rectal palpation a growth was palpable in the apex of one uterine horn. The growth was removed by right flank laparotomy under sedation and paravertebral nerve block. The growth was diagnosed to be a fibroma. The cow conceived and calved normally after the operation. PMID- 15119787 TI - Occurrence and pathology of Sinergasilus lieni (Copepoda: Ergasilidae), a parasite of the silver carp and bighead, in Hungarian ponds. AB - Sinergasilus lieni Yin, 1949, a well-known and pathogenic parasitic copepod in China and Russia, has been detected in Hungarian carp farms for the first time. The parasite infected the third-year generation of silver carp (Hypophthalmichthys molitrix) and bighead (Aristichthys nobilis). The gills of the infected fish specimens showed severe pathological changes. At the attachment sites of female copepods clubbing and fusing of the gill filaments were observed and in some parts of the pale or whitish hemibranchia deep indentations were recorded in places where the tips of the damaged filaments had broken off. Silver carp and bighead were infected at a similar rate, having 8 to 27 copepods attached to the end of the clubbed filaments or the proliferated epithelium of 2 to 10 fused filaments. In histological sections the head part of the parasite was found in a deep cavity of the proliferated epithelium, piercing its antennae deep into the tissues. Only the end of the filaments showed changes. In this part the proliferated epithelium was infiltrated by eosinophilic granular cells. In the central and basal parts of the hemibranchia the original structure of the filaments was preserved with intact secondary lamellae. PMID- 15119788 TI - Immune response in mice infected by Encephalitozoon cuniculi and suppressed by dexamethasone. AB - Several indicators of immune response were observed in immunocompetent mice of the ICR line and those suppressed by dexamethasone upon their experimental infection with the microsporidia of Encephalitozoon cuniculi. The mice were infected by one-shot intraperitoneal administration of 5 x 10(7) pathogenic spores. On Days 7, 14, 28 and 42 after infection, peripheral blood leukocyte phagocytic activity was determined and compared, including phagocytic index and the blastogenic response in spleen cells to mitogenic activation by concanavalin A and phytohaemagglutinin. The results point to the fact that E. cuniculi itself can cause a significant decrease in phagocytic activity of phagocytic leukocytes in the early stages of infection as well as a remarkable decrease in the proliferative response of spleen cells to T-cellular mitogens. PMID- 15119789 TI - Immunohistochemical detection of fungal elements in the tissues of goslings with pulmonary and systemic aspergillosis. AB - Nineteen goslings with pulmonary and systemic aspergillosis were the subject of the study. The lungs and air sacs were the main sites affected by the disease, and were generally characterised by diffuse yellowish-white granulomas. In 7 cases with pulmonary and air-sac involvement the granulomas were scattered to the serosal linings of the gastrointestinal and upper respiratory tracts, to the liver, spleen and kidneys, and in two cases also to the bursa of Fabricius, musculus (m.) longus colli and adventitia of aorta. The granulomas were often characterised by a necrotic centre surrounded by heterophils, macrophages, lymphocyte and plasma cells, and in late granulomas by multinucleated foreign body giant cells, and again by an outer thin fibrous capsule. Numerous fungal hyphae were found within the necrotic debris of the granulomas by Gridley and PAS staining techniques. Immunohistochemistry reliably confirmed aspergillosis in all of the cases. Fungal elements in the lungs of goslings severely affected by the disease stained heavily within the centre of the granulomas, whereas few antigens reacted in the chronic cases. Fungal fragments, which were not discernible using routine fungal stains, reacted clearly in the cytoplasm of macrophages and giant cells. Thus, although fungal elements within the granulomas were histologically indicative of aspergillosis, immunohistochemistry also had to be applied to obtain a definitive diagnosis of the disease and to differentiate it from many of the filamentous fungi. PMID- 15119790 TI - Pharmacokinetics and clinical efficacy of cefotaxime for the treatment of septicaemia in dogs. AB - Considering the already known pharmacological features of cefotaxime, a study with two approaches of pharmacokinetics and clinical efficacy in septicaemic dogs was carried out. Pharmacokinetic variables were defined for doses of 10 mg/kg, and 20 mg/kg, utilising a quantitative bacteriological analysis. Values for half life (T1/2 beta) at 10 mg/kg were 0.8, 1.48 and 1.52 h for the i.v., s.c. and i.m. routes, respectively. Corresponding values for the 20 mg/kg dose for the same routes were 0.8, 1.49 and 1.53 h, respectively. Relatively fast clearance (ranging from 0.58 to 0.64 L/kg/h) allowed a maximum dose interval of 12 h. The above-stated doses of cefotaxime were administered i.v. to 40 cases of septicaemia, clinically divided into 20 moderately severe cases treated with 10 mg/kg i.v., of cefotaxime bid, and 20 severe ones, treated with 20 mg/kg i.v. of cefotaxime bid. Injections continued until a previously defined criterion of 'clinically recovered' was obtained. Thereafter, a follow-up treatment was established using the same dose and dose-interval but through the s.c. route. Due to the apparent volumes of distribution obtained (ranging from 0.48 to 0.51 L/kg), considering the overall clinical efficacy obtained (90% for the 10 mg/kg dose and 75% for the 20 mg/kg dose), and due to the rapid improvement observed after a few doses of the drug (1.8 to 2.5 doses to 'clinical improvement'), it is safe to postulate such doses of cefotaxime as excellent choices for the treatment of septicaemia in dogs. PMID- 15119791 TI - Expression and localisation of leptin and leptin receptor in the mammary gland of the dry and lactating non-pregnant cow. AB - Leptin and leptin receptor were studied in the mammary gland of non-pregnant dry and lactating cows. Using RT-PCR it was demonstrated that leptin and its short (Ob-Ra) and long (Ob-Rb) receptor isoforms are expressed both in the dry and the lactating mammary gland tissue. Tissue distribution of leptin and its receptor mRNA transcripts were examined by in situ hybridisation, while the leptin protein was localised by immunohistochemistry. Although in situ hybridisation is semiquantitative, our morphological data suggest that the epithelial leptin mRNA expression of the lactating gland is higher than that of the dry gland. To compare the leptin mRNA levels between dry and lactating udders competitive PCR was used, which showed no difference in leptin expression for the whole mammary tissues. The lack of difference in total leptin mRNA levels is explained by the high adipose tissue content of the dry mammary gland. Leptin and its receptor transcripts are expressed mainly in the epithelial cells of lactating cows, while in dry mammary tissue the signal is found in the stromal tissues as well. The results provide additional evidence that locally produced leptin takes part in the regulation and maintenance of mammary epithelial cell activity. PMID- 15119792 TI - Assisted reproductive research: laser assisted hatching and spindle detection (spindle view technique). AB - Animal experiments are very important for the development of new assisted reproductive techniques (ART) for use in human and animal reproductive medicine. Most technical aspects of reproductive manipulation of humans and animals are very similar, and many components of successful human ART used nowadays have been derived from animal studies. In this study we examined (1) the use of 'non contact' laser for assisted hatching, (2) whether spindles in living mouse oocytes could safely be imaged/examined by polarisation microscope (polscope) and (3) the influence of environment (e.g. temperature, in vitro culture, etc.) on spindle detection/visualisation. The data of the study presented here show that (1) laser assisted hatching (AH) is a fast, very accurate and safe procedure without any harmful effect on embryo development and it can support very effectively the implantation of embryos, (2) the use of polscope facilitates the evaluation of oocyte quality and the selection of oocytes with spindle, (3) by monitoring the spindle position during intracytoplasmic sperm injection (ICSI), we can reduce spindle damage and increase the chance of fertilisation. Further studies are underway to test the hypothesised connection between spindle birefringence and developmental capacity of oocytes/embryos. PMID- 15119793 TI - Determination and phylogenetic analysis of canine distemper virus in dogs with nervous symptoms in Turkey. AB - In the present study, canine distemper virus (CDV) was investigated in 20 dogs having nervous signs arousing the clinical suspicion of canine distemper (CD). A total of 13 animals (65%) were stray dogs and had no accurate record about the vaccination history. Clinical examinations revealed that the majority (85%) of the animals showed systemic form characterised by predominantly nervous symptoms accompanied by mild respiratory system signs whilst the remaining cases (15%) recorded mainly respiratory distress. CDV RNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) only in 45% of the suspected cases. Phylogenetic analysis of partial nucleotide sequence of the P gene coding region revealed that the virus is closely related to European strains. Immune responses in 13 cases (65%), which were detected by dot-ELISA, indicated inefficient levels for neutralising functions against CDV. It was postulated that this response could have been mediated by either previous vaccination or mild infection with field strains. PMID- 15119794 TI - Canadian dentists' willingness to be involved in dental research. AB - BACKGROUND: Building a collaborative research network reuniting dentists and academics constitutes a solution in order to bridge the gap between dental research and patient care. The purpose of this study was to identify the kind of clinicians willing to be involved in research and to determine their research priorities. METHODS: A questionnaire was mailed to all registered dentists in Canada in December 2001. This questionnaire comprised sociodemographic variables and questions on research utility, research results availability and dental research priorities. The statistical analyses were performed with 2,595 questionnaires completed by dentists working in a clinical setting. RESULTS: 27% of respondents were willing to be involved in dental research, 23% did not know and 50% did not want to be involved. A multiple logistic regression model shows that being open to participate (Yes and Don't know) is associated with: younger age (OR = 2.83), perception that research has a very big impact on the oral health of the population (OR = 1.93), perception that dental research results are not easily available to dentists (OR = 1.47), practice as a specialist (OR = 1.45) and French spoken as a first language (OR = 1.45). A large majority (80%) of dentists who would like to be involved in research think that effectiveness of techniques and treatments are a very high priority. CONCLUSIONS: There is a significant group of dentists who wish to be involved in research. This information could be used to reunite dentists and researchers in a collaborative network. PMID- 15119795 TI - Comparison of the dental education systems in Korea and Japan. AB - In recent years, international relationships in dentistry have grown stronger through journals, books, academies, seminars, researcher exchanges and so forth. Korea and Japan are neighbours in East Asia. However, no comparison of the dental education systems of the two countries has been published. Therefore, the authors have provided the present comparison to promote mutual understanding and to familiarise dentists around the world with dental education in these two countries. The number of dentists, life expectancy at birth, and number of decayed, missing, and filled teeth (DMFT) at age 12 years in Korea and Japan are summarised. PMID- 15119796 TI - Periodontal treatment needs in a native island community in Colombia determined with CPITN. AB - AIMS: To identify the prevalence and different degrees of periodontal disease in an isolated community (Isla Grande, Colombia) with no dental services and low educational level with the use of CPITN, and to establish periodontal treatment needs in different age groups. RESULTS: Of 116 people examined, 0.9% were in periodontal health (CPITN value 0), 18.1% had gingival bleeding (CPITN value 1), 51.7% had supra or subgingival calculus (CPITN value 2), 18.1% presented pockets 3.5-5.0 mm deep (CPITN value 3), and 11.2% had pathological pockets of 5.5mm or deeper (CPITN value 4). No clear differences were observed between sexes. CONCLUSIONS: This study shows that 81% of the sample has some type of periodontal treatment need, with 69.8% of them requiring periodontal treatment that may be supplied by a hygienist and 11.2% requiring specialised treatment. Implementation of oral health education and oral prevention programmes was recommended to the authorities for this community. PMID- 15119797 TI - Epidemiological aspects of oral tori in a Ghanaian community. AB - AIM: To determine the prevalence of oral tori, commonly found among Ghanaians, and compare with that in other regions. METHOD: Dental examination records of all 926 patients seen from January 1998 to April 2000 in a dental outpatient clinic in Accra, Ghana, were studied. The existence of a torus had systematically and routinely been ascertained by visual inspection and palpation. RESULTS: Frequency distribution and cross-tabulation analysis showed an overall prevalence of 14.6% with a female: male ratio of 1.1: 1. The most prevalent variety was the bilateral mandibular torus (12.1%) and the midpalatal torus of the maxillary tori (4.3 %). Females had 2.2 times the probability of having midpalatal torus compared to men (Odds Ratio (OR) = 2.2; Confidence Interval: 1.05, 4.70). There was a very strong concurrent relationship between mandibular and maxillary tori (OR = 16; CI = 7.8, 32.5). CONCLUSION: Comparisons indicated a strong similarity between torus prevalence in Ghana and the Caribbean regions. This should help in further discussions on the epidemiology of this bony anomaly. PMID- 15119798 TI - Oral health status of children and adults in urban and rural areas of Burkina Faso, Africa. AB - OBJECTIVES: To analyse the oral health status of children and adults in rural and urban areas of Burkina Faso; to provide epidemiological data for planning and evaluation of oral health care programmes. DESIGN: Cross-sectional survey including different ethnic and socio-economic groups. SAMPLE AND METHODS: Multistage cluster sampling of households in urban areas and random samples of participants selected based on the recent population census in rural areas. The final study population covered four age groups: 6 years (n = 424), 12 years (n = 505), 18 years (n = 492) and 35-44 years (n = 493). Clinical oral health data collected according to WHO methodology and criteria. RESULTS: At age 6, 38% of children had caries, with prevalence higher in urban than rural areas. At age 12, the mean DMFT was 0.7 with prevalence significantly higher among urban than rural children. Mean DMFT was 1.9 in 18-year-olds and 6.3 in 35-44-year-olds and figures were higher for women than men. In adults, no differences in caries experience were found by location whereas the caries index was significantly affected by ethnic group and occupation. CPI score 2 (gingivitis and calculus) was dominant for all ages: 6 years (58%), 12 years (57%), 18 years (58%), 35-44 years (49%). In addition, 10% of 35-44-year-olds had CPI score 4. Rural participants had more severe periodontal scores than did urban individuals. CONCLUSIONS: Health authorities should strengthen the implementation of community based oral disease prevention and health promotion programmes rather than traditional curative care. PMID- 15119799 TI - Utilisation of dental services in Turkey: a cross-sectional survey. AB - AIM: To assess the factors affecting the utilisation of dental service in Turkey. METHOD: A cross-sectional study using a sample of 866 adults randomly selected from nine provinces, collecting data on socio-demographic characteristics, frequency of dental visits and attitudes toward dental services in face-to-face interviews. RESULTS: 40.4% of adults had visited a dentist within the previous year. Logistic regression analysis showed that gender and educational level were significant factors affecting the utilisation of dental services, with women using dental services significantly more than men and those having higher education compared to those having low education. Satisfaction from the services of the private dental offices and private hospitals was higher than from state hospitals. Age, reasons for dental visit and the place of the last visit were found to be the most significant predictors of satisfaction. CONCLUSION: Utilisation of dental services is low amongst Turkish adults. Women and individuals with higher levels of education use dental services more than others. The majority of the people prefer private dental offices, where the patient satisfaction is high. PMID- 15119800 TI - Odontogenic infections and descending necrotising mediastinitis: case report and review of the literature. AB - Descending necrotising mediastinitis is a rare complication secondary to oral surgery or odontogenic infections which ultimately spreads to the mediastinum through the anatomical cervical spaces. Delay in diagnosis, despite broad spectrum antimicrobical therapy and extensive surgical intervention, results in an unacceptably high mortality rate (almost to 40%). A case study is presented as a rationale for a review of the current literature and to analyse the evolution in therapies, both medical and surgical from Pearse in 1938 to today. PMID- 15119801 TI - Herpes simplex virus: an occupational hazard in dentistry. AB - Approximately 70% of the population in the western world become infected with the herpes simplex virus type 1 (HSV-1) by the second decade of life. This review discusses the role of the HSV-1 as a potential occupational hazard for dental workers, focusing on herpes labialis, herpetic whitlow and keratitis. The risks associated with the dental treatment of patients with HSV-1, both from the perspective of the clinician and the patient are presented. Procedures for minimising the impact of HSV-1 within the dental practice, in particular infection control, delivery of treatment and patient education, are addressed. The management options for recurrent herpes labialis are also reviewed. PMID- 15119802 TI - A warm welcome to the occupational health: Spanish column. PMID- 15119803 TI - A warm welcome to the occupational health: Spanish column. PMID- 15119804 TI - A warm welcome to the occupational health: Spanish column. PMID- 15119805 TI - A warm welcome to the occupational health: Spanish column. PMID- 15119806 TI - BMI is not a magic number. PMID- 15119807 TI - BMI is not a magic number. PMID- 15119808 TI - Disabler article. PMID- 15119809 TI - AAOHN Code of Ethics and interpretative statements. PMID- 15119810 TI - What are some of the challenges in case management and how have you handled them? PMID- 15119811 TI - Occupational and environmental exposure history. PMID- 15119812 TI - Ensuring confidentiality of employee health information--developing policies and procedures. PMID- 15119813 TI - Environmental health hazards and health care professional education. PMID- 15119814 TI - Meniere's disease: identifying classic symptoms and current treatments. AB - Meniere's disease affects 615,000 individuals in the United States. Therefore, occupational health nurses may encounter employees who express symptoms of Meniere's disease or have been diagnosed with this problem. When nurses have an understanding of the symptoms of Meniere's disease, they can validate clients' concerns by directing them to an otolaryngologist or audiologist for further evaluation. Awareness of Meniere's disease may aid in counseling individuals during their annual education and training that is required to maintain an effective hearing conservation program. PMID- 15119815 TI - The new face of public health in the workplace: development of an occupational health program. AB - Public health nursing systems can provide essential occupational health services to the private sector. New occupational health programs can be created by public health agencies for private sector organizations without onsite occupational health programs. The Centers for Disease Control and Prevention for Implementing and Evaluating Public Health Systems provide an excellent framework for development of targeted programs to meet employee health needs. Outsourcing of occupational health services by private businesses and government agencies to their local health department can be effective and economical. Innovative entrepreneurial efforts by health department nurses can bring direct interventions into occupational settings to improve the health of the individual, family, and community and thus reduce the public health burden. PMID- 15119816 TI - Monkeypox in the United States: an occupational health look at the first cases. AB - Between May 15 and June 20, 2003, 71 suspected cases of monkeypox were investigated and 37 individuals in the United States developed laboratory confirmed monkeypox. These were the first cases of human monkeypox ever documented in the United States or in the Western Hemisphere. The disease was transmitted from small animals imported from Africa to other animals, including prairie dogs sold as pets throughout the U.S. Midwest. Direct contact with the infected animals was the method of infection, and although human to human transmission was thought to have occurred, this was not confirmed by follow up testing. Because of the link with contact with a prairie dog, initial evaluation of the disease was focused toward diseases commonly associated with this animal (e.g., tularemia, plague). Laboratory findings at the Marshfield Clinic in Marshfield, Wisconsin pointed to the presence of an orthopox. The CDC confirmed monkeypox was the infecting orthopox agent. Occupational health nurses from the Marshfield Clinic had direct involvement in the identification and follow up of employees who had direct contact with the diagnosed patients. Programs, such as a respiratory protection program initiated and carried out by Clinic occupational health nurses, were used to prevent employee exposure for Clinic staff. One Clinic employee was thought to potentially have monkeypox because of her direct contact with one of the patients. Four Clinic employees were vaccinated with vaccinia vaccine as a result of their contact with patients or lab specimens. Quarantine of the potentially infected employee and her boyfriend uncovered issues that must be addressed if other infectious diseases requiring quarantine or isolation of individuals emerge or re-emerge. These include a system to compensate individuals in quarantine or isolation who do not have any other source of income. The issue of whether workers' compensation should cover an employee who is quarantined or isolated for a potential work related exposure to an infectious disease if no disease is actually diagnosed also needs to be explored. A better system of getting state or CDC laboratory results back to the local level, including the occupational health area of the generating facility, must be developed. This will be very important if diseases such as severe acute respiratory syndrome (SARS) or smallpox should re-emerge in the United States. Occupational health nurses are an integral part of any infectious disease process occurring in the United States. The identification of monkeypox in the United States shows that any planning to detect, prevent, and treat diseases with the potential to affect the employee population must include occupational health nurse involvement. PMID- 15119817 TI - Roadmap to HIPAA: keeping occupational health nurses on track. AB - So what does HIPAA require most covered entities to do? At this point, the Privacy Rule compliance date has already passed for all covered entities except small health plans. Most of the requirements under the Privacy Rule dictate the development of appropriate policies and procedures, a notice of privacy practices and other forms, implementation of measures to secure the privacy of PHI, contracting with Business Associates, and training of all involved. For covered entities, testing of the electronic standard transactions to exchange data between participating parties should have begun by April 16, 2003. Although full implementation of the electronic transactions should have taken place by October 16, 2003, the government has allowed covered entities that are still actively working toward compliance to operate under contingency plans. It remains unclear when the use of such plans will be disallowed. After standards are published for claim attachments and first report of injury, these electronic standard transactions will be incorporated by the designated compliance date. Appropriate use of national identifiers will be implemented after final rules and standards are published. For the occupational health nurse who is not a covered entity, the most critical implementation factor is a HIPAA compliant authorization form so the occupational health nurse can continue to obtain necessary PHI. This is essential when attempting to obtain medical information, even for workers' compensation or disability case management. Although these plans are not considered health plans under HIPAA and, therefore, would not require the designation of covered entity, the occupational health nurse frequently needs to obtain PHI to manage these cases. Most providers in the health care community will be covered entities under HIPAA and will not be able to release PHI without a signed HIPAA compliant authorization form. In addition, providers will want a HIPAA compliant authorization form signed when requesting health information from the occupational health nurse. The HIPAA's privacy regulations are considered "the floor" or minimum standard for the protection of PHI. As such, it is likely that these privacy regulations will become the "industry standard" to which all health care professionals will be held. Even though the occupational health nurse may not be a covered entity, implementing appropriate HIPAA procedures is recommended. Knowing that most of HIPAA's privacy rule contains requirements already in place and in practice for most occupational health nurses can take some of the worry out of this complex regulation. Additionally, the nurse interacts with the health care system in a variety of roles. As a health care consumer, occupational health nurses can assert their own patient rights when interacting with covered entities. As the trusted advisor and consultant to many employees, the occupational health nurse can play a vital role in educating employees about HIPAA and assisting employees with navigating an ever-complex health care system. As a health care professional, the occupational health nurse continues to protect and safeguard all PHI while respecting employees' rights and delivering quality care. Staying knowledgeable and up-to-date on the HIPAA regulations as they continue to evolve and change allows occupational health nurses to stay on the right course while mapping their way toward regulatory compliance (see Sidebar for recommended resources). PMID- 15119818 TI - What works for weight loss and maintenance? PMID- 15119819 TI - Molecular insights into mental retardation: multiple functions for the Fragile X mental retardation protein? AB - Mental retardation is a frequent cause of intellectual and physical impairment. Several genes associated with mental retardation have been mapped to the X chromosome, among them, there is FMR1. The absence of or mutation in the Fragile Mental Retardation Protein, FMRP, is responsible for the Fragile X syndrome. FMRP is an RNA binding protein that shuttles between the nucleus and the cytoplasm. FMRP binds to several mRNAs including its own mRNA at a sequence region containing a G quartet structure. Some of the candidate downstream genes recently identified encode for synaptic proteins. Neuronal studies indicate that FMRP is located at synapses and loss of FMRP affects synaptic plasticity. At the synapses, FMRP acts as a translational repressor and in particular regulates translation of specific dendritic mRNAs, some of which encode cytoskeletal proteins and signal transduction molecules. This action occurs via a ribonucleoprotein complex that includes a small dendritic non-coding neuronal RNA that determines the specificity of FMRP function via a novel mechanism of translational repression. Since local protein synthesis is required for synaptic development and function, this role of FMRP likely underlies some of the behavioural and developmental symptoms of FRAXA patients. Finally we review recent work on the Drosophila system that connects cytoskeleton remodelling and FMRP function. PMID- 15119820 TI - Cancer gene suppression strategies: issues and potential. AB - Oncogenes are ideal targets for therapies which down-regulate gene expression. However, effective modalities for altering gene expression in vivo have thus far proven to be elusive. Whilst there has been recent success with small molecule inhibitors of oncoprotein function, evolution of resistance to these agents has been observed in the clinical setting, indicating the need for combinations of therapies for cancer treatment. Strategies for in vivo gene down-regulation still hold promise for the treatment of cancer. The technologies relevant to such therapeutic strategies are discussed in terms of molecular action, delivery and choice of target gene. Consideration is given to the pre-clinical and clinical efficacy these agents have demonstrated to date. PMID- 15119821 TI - Precise circadian clocks in prokaryotic cyanobacteria. AB - Prokaryotic cyanobacteria express robust circadian (daily) rhythms under the control of a timing mechanism that is independent of the cell division cycle. This biological clock orchestrates global regulation of gene expression and controls the timing of cell division. Proteins that may be involved in input pathways have been identified. Mutational screening has identified three clock genes that are organized as a gene cluster. The structure of cyanobacterial clock proteins, their phosphorylation, and regulation is described. A new model for the core clockwork in cyanobacteria proposes that rhythmic changes in the status of the chromosome underlie the rhythms of gene expression. Mixed-strain experiments demonstrate that this timekeeper confers adaptive value when different strains compete against each other. PMID- 15119822 TI - Type V protein secretion: simplicity gone awry? AB - Since its discovery in the late 1980's, the family of secreted proteins termed the autotransporters has been expanding continuously to become the largest group of secreted proteins in Gram-negative bacteria. The type V secretion pathway, which includes the autotransporters (type Va) together with the two-partner secretion system (type Vb) and the Oca family (type Vc), can be defined by secreted proteins that are (i) translocated across the outer membrane via a transmembrane pore formed by a beta-barrel and (ii) contain all the information required for translocation through the cell envelope. In the light of new discoveries and controversies in this research field, the secretion process of autotransporters, or the type Va secretion system, will be discussed here and placed in the context of the more general field of bacterial protein translocation. PMID- 15119823 TI - Recent developments in bacterial cold-shock response. AB - In response to temperature downshift, a number of changes occur in cellular physiology such as, (i) decrease in membrane fluidity, (ii) stabilization of secondary structures of nucleic acids leading to reduced efficiency of mRNA translation and transcription, (iii) inefficient folding of some proteins, and (iv) hampered ribosome function. Cold-shock response and adaptation has been quite extensively studied in Escherichia coli and Bacillus subtilis. A number of cold shock proteins are induced to counteract these harmful effects of temperature downshift. General principles of cold-shock response along with recent findings on desaturase system, RNA chaperone and transcription antitermination function of CspA homologues, cold shock induction of chaperones and synthesis of trehalose, CspA homologues from hyperthermophilic bacteria and possible multiple roles of cold shock proteins in other stress responses of bacteria are discussed. PMID- 15119824 TI - Stability and repair of DNA in hyperthermophilic Archaea. AB - Evolutionary and physiological considerations argue that study of hyperthermophilic archaea should reveal new molecular aspects of DNA stabilization and repair. So far, these unusual prokaryotes have yielded a number of genes and enzymatic activities consistent with known mechanisms of excision repair, photo-reversal, and trans-lesion synthesis. However, other DNA enzymes of hyperthermophilic archaea show novel biochemical properties which may be related to DNA stability or repair at extremely high temperature but which remain difficult to evaluate rigorously in vivo. Perhaps the most striking feature of the hyperthermophilic archaea is that all of them whose genomes have been sequenced lack key genes of both the nucleotide excision repair and DNA mismatch repair pathways, which are otherwise highly conserved in biology. Although the growth properties of these micro-organisms hinder experimentation, there is evidence that some systems of excision repair and mutation avoidance operate in Sulfolobus spp. It will therefore be of strategic significance in the next few years to formulate and test hypotheses in Sulfolobus spp. and other hyperthermophilic archaea regarding mechanisms and gene products involved in the repair of UV photoproducts and DNA mismatches. PMID- 15119825 TI - The molecular biology of recombination in Mycobacteria: what do we know and how can we use it? AB - Recombination is a ubiquitous genetic process which results in the exchange of DNA between two substrates. Homologous recombination occurs between DNA species with identical sequence whereas illegitimate recombination can occur between DNA with very little or no homology. Site-specific recombination is often used by temperate phages to stably integrate into bacterial chromosomes. Characterisation of the mechanisms of recombination in mycobacteria has mainly focussed on RecA dependent homologous recombination and phage-directed site-specific recombination. In contrast the high frequency of illegitimate recombination in slow-growing mycobacteria has not been explained. The role of DNA repair in dormancy and infection have not yet been fully established, but early work suggests that RecA-mediated pathways are not required for virulence. All three recombination mechanisms have been utilised in developing genetic techniques for the analysis of the biology and pathogenesis of mycobacteria. A recently developed method for studying essential genes will generate further insights into the biology of these important organisms. PMID- 15119826 TI - Molecular biology of microbial hydrogenases. AB - Hydrogenases (H2ases) are metalloproteins. The great majority of them contain iron-sulfur clusters and two metal atoms at their active center, either a Ni and an Fe atom, the [NiFe]-H2ases, or two Fe atoms, the [FeFe]-H2ases. Enzymes of these two classes catalyze the reversible oxidation of hydrogen gas (H2 <--> 2 H+ + 2 e-) and play a central role in microbial energy metabolism; in addition to their role in fermentation and H2 respiration, H2ases may interact with membrane bound electron transport systems in order to maintain redox poise, particularly in some photosynthetic microorganisms such as cyanobacteria. Recent work has revealed that some H2ases, by acting as H2-sensors, participate in the regulation of gene expression and that H2-evolving H2ases, thought to be involved in purely fermentative processes, play a role in membrane-linked energy conservation through the generation of a protonmotive force. The Hmd hydrogenases of some methanogenic archaea constitute a third class of H2ases, characterized by the absence of Fe-S cluster and the presence of an iron-containing cofactor with catalytic properties different from those of [NiFe]- and [FeFe]-H2ases. In this review, we emphasise recent advances that have greatly increased our knowledge of microbial H2ases, their diversity, the structure of their active site, how the metallocenters are synthesized and assembled, how they function, how the synthesis of these enzymes is controlled by external signals, and their potential use in biological H2 production. PMID- 15119827 TI - Molecular phylogeny in 3-D. AB - Molecular phylogenetic trees are constructed in three dimensions relative to the distribution of MW and pl classes and immunocrossreactivity against polyclonal antibodies to lens crystallins, as well as multiple sequence alignment between amino acid sequences, coding nucleotide sequences and the gene nucleotide sequences for beta-globin. Euclidian distances are estimated to position species in x, y, z space by multidimensional scaling and merged with bootstrap-tested branching pattern of Fitch & Margoliash plots to obtain 3-D phylogenetic tree. Compared to single attributes, phylogenetic trees based on multiple parameters allow significant repositioning of rodents, chiroptera and primates. PMID- 15119828 TI - Helicobacter pylori infection in Finland. AB - Helicobacter pylori causes chronic gastritis worldwide and it is the most important single factor in peptic ulcer disease. Up to half of H. pylori infected individuals develop atrophic gastritis over years and decades. H. pylori infection has also been classified as a class I carcinogen in human gastric cancer. Most infections are obtained in childhood, in Finland mainly before the age of 7 years but the exact transmission routes are not known. The infection shows an age-dependent pattern, the infection being rare among children but gradually becoming more prevalent among older age groups. As new infections are few in adults and the infection only rarely disappears without effective anti microbial therapy, the occurrence of the infection in the old actually reflects the prevalence of the infection in their childhood. In developed countries, such as Finland, a rapid decline of H. pylori prevalence rate has been demonstrated. In order to speed up this natural decline of the infection, a unique population based 'screen and treat' project was started in Vammala, a semiurban south western community in Finland. In this survey, young inhabitants were offered diagnosis and treatment for H. pylori. PMID- 15119829 TI - Molecular basis of Mendelian idiopathic epilepsies. AB - A genetic aetiology is estimated to be present in about 40% of patients with epilepsy. Significant progress has been made in understanding the molecular genetic basis of Mendelian epilepsies. Fourteen genes have been identified which underlie a group of rare, autosomal dominant Mendelian idiopathic epilepsies. All but two of these genes encode subunits of ion-channels, revealing that idiopathic Mendelian human epilepsies are predominantly channelopathies. The two non-ion channel genes, LGl1 causing autosomal dominant lateral temporal lobe epilepsy and MASS1 causing febrile and afebrile seizures, both contain a novel repeat motif variously called the epilepsy-associated repeat (EAR) and epitempin (EPTP) repeat. This motif defines a subfamily of genes, some of which have also been implicated in epilepsy in mice and humans. Progress in dissecting the more common 'complex' genetic epilepsies remains slow, but ion channels represent the most biologically plausible candidates. Characterization of common population sequence variants for the entire cohort of ion channel genes and the development of high throughput techniques should enable rapid advances in the understanding of the common idiopathic familial epilepsies. PMID- 15119830 TI - Chemokines and atherosclerosis. AB - Atherosclerosis is an inflammatory disease of the vessel wall, characterized by the accumulation of leukocytes, especially macrophages and T-cells. Chemokines are small heparin-binding polypeptides, whose main function is to attract cells to the areas of developing inflammation. They function by ligating G-protein coupled chemokine receptors initiating different signaling cascades. In vivo and in vitro investigations showed that chemokines are produced by a variety of cells and play important roles in the development and progression of many physiological and pathological conditions including atherosclerosis. Chemokines such as MCP-1, MCP-4, MIP-1 and RANTES may mediate leukocyte trafficking to, and their retention in, the plaque while CXCL16 seems to fulfill the dual function of a chemokine and a scavenger receptor. Chemokine and chemokine receptor homologues are secreted by several viruses, which may also play a role in the pathogenesis of atherosclerosis. Expression levels and gene polymorphisms of some chemokines may become useful clinical markers of atherosclerosis and other cardiovascular diseases. Modulation of chemokines and chemokine receptors' expression as well as their signaling pathways may provide important anti-atherogenic strategies. PMID- 15119831 TI - Thrombogenesis, atherogenesis and angiogenesis in vascular disease: a new 'vascular triad'. AB - Amongst the components of the pathophysiology of atherosclerosis are the tendency to thrombus formation (i.e., thrombogenesis) and the loss of endothelial cell integrity. The endothelium is also implicated in atherogenesis, as vasa vasorum are present in the adventitia and media at a higher density in atherosclerotic tissue, and that neovascularisation leading to collateral growth bypassing obstruction and/or stenoses provides important alternative routes to feeding distal tissues. The development of these new blood vessels is the process of angiogenesis. The present 'viewpoint' article will explore the relationships between the three processes. There is considerable evidence that atherogenesis and thrombogenesis are intimately linked, but angiogenesis in cardiovascular disease is a recently developed concept. We propose a new 'vascular triad' of these abnormal pathophysiological processes, leading to an alternative view of the pathogenesis of vascular disease. Indeed, we suggest that angiogenesis is an important aspect of the pathophysiology of cardiovascular disease, impacting with both thrombogenesis and atherogenesis, in a new vascular triad (the 'Birmingham vascular triad'). PMID- 15119832 TI - Plant stanol and sterol esters in prevention of cardiovascular diseases. AB - Statin trials have indicated that effective reduction of serum cholesterol should last up to one year before reduced risk of cardiovascular diseases can be detected. This observation can be applied most probably also to the use of plant stanol/sterol ester spreads for the treatment of hypercholesterolemia. However, despite the fact that the two spreads lower serum cholesterol similarly in short term studies, a comparison of one year results reveals an inconsistent effect of plant sterol spread as compared with that of plant stanol spread on cholesterol concentration in both men and women. This favors the use of plant stanol ester spread for long-term lowering of serum cholesterol. Doses of about 2 g/day of plant stanols as fatty acid ester spread enhances fecal elimination of cholesterol, but not of bile acids, through inhibition of cholesterol absorption by about 40%. This lowers serum total and low density lipoprotein (LDL) cholesterol despite enhanced compensatory increase in cholesterol synthesis by about 10% and 15% as compared with control spread, respectively, and by up to 20% as compared with the baseline diet. About one-third of mildly hypercholesterolemic subjects reach an accepted cholesterol level. A small dose of statin should be added to treatment in individuals resistant to monotherapy with plant stanol ester spread. A life-long consumption of plant stanol ester spread has been predicted to lower coronary events by about 20%. PMID- 15119833 TI - The ins and outs of reverse cholesterol transport. AB - It is generally assumed that HDL is the obligate transport vehicle for 'reverse cholesterol transport', the pathway for removal of excess cholesterol from peripheral tissues via the liver into bile and subsequent excretion via the feces. During the last few years, intensive research has generated exciting new data on the separate processes involved in reverse cholesterol transport. Many 'new' proteins, particularly members of the ABC transporter and nuclear receptor subfamilies, that mediate or influence cholesterol fluxes have been identified and characterized. An important role of the intestine in regulation of cholesterol homeostasis is emerging. In this paper, new insights into mechanisms of reverse cholesterol are reviewed. PMID- 15119834 TI - DNA pooling as a tool for large-scale association studies in complex traits. AB - Pooled genotyping is a powerful and efficient tool for high throughput association analysis, both case-control and family-based. The use of pooling designs can significantly reduce the consumable and labour costs of a study. At the same time, since it is also extremely efficient with DNA resources, pooling can be an extremely effective method for conserving DNA. Here, we consider the main points in study design, and illustrate the application of pooling using psychiatric phenotypes as an example. PMID- 15119835 TI - The white blood cell count is an independent predictor of no-reflow and mortality following acute myocardial infarction in the coronary interventional era. AB - BACKGROUND: In the era before the use of coronary reperfusion therapy, an elevated white blood cell (WBC) count was associated with a higher risk of adverse events following acute myocardial infarction (AMI). However, the relationship between WBC count and prognosis after AMI has not been investigated since coronary intervention was introduced. AIM: To evaluate whether a high WBC count within 48 hours of the onset of AMI predicts future adverse events in patients undergoing percutaneous coronary intervention (PCI). METHOD: We evaluated 1,016 patients who underwent PCI in the acute phase of MI using the Japanese Acute Coronary Syndrome Study (JACSS) database. RESULTS. WBC count was significantly associated with smoking, sudden onset AMI, and the no-reflow phenomenon during PCI, as were age, peak creatine kinase level, and Killip class. An elevated WBC count was significantly associated with higher risk of in hospital mortality. Patients in the highest quartile of WBC count were about three times more likely to have a poor prognosis after AMI compared to those in the lowest quartile. CONCLUSIONS: The WBC count is of great significance for stratifying patient risk and can be used as a universal marker for predicting future adverse events following any treatment for AMI. PMID- 15119836 TI - Library involvement in the implementation of a course management system. AB - Course management system software has much to offer the faculty and students in universities that adopt this technology. Libraries have used this technology to provide links to library resources, but they can also have a role in implementing the adoption of the course management software. The Penn State College of Medicine successfully implemented the ANGEL course management system under the guidance of a medical librarian. Tasks specific to the librarian included communication and promotion of ANGEL, in addition to providing training and technical support. PMID- 15119837 TI - Asked and answered--online: how two medical libraries are using OCLC's QuestionPoint to answer reference questions. AB - This paper imparts the experiences of two similar but unaffiliated medical libraries that use QuestionPoint, OCLC's collaborative virtual reference product. The authors introduce the major features of QuestionPoint, with particular emphasis on its asynchronous e-mail reference service. After presenting how both libraries have employed this service in their respective environments, the paper examines the quantity and quality of reference questions submitted via QuestionPoint. The types of questions are explicated, and statistical trends are compared. PMID- 15119838 TI - The library as finals resting place: expanding service to health sciences students during final exams. AB - To increase services to health sciences students during their final exam period, and to demonstrate to campus administration that the library is in tune with the students' fluctuating needs, the Kornhauser Health Sciences Library, University of Louisville, increased its hours of operation; created an inviting, comfortable environment; and offered free snacks and drinks for the students. The event, coined the "Finals Resting Place," was a positive public relations tool that strengthened the library's relationship with its students. Moreover, it reinforced the library's role and mission to the campus administration, particularly that of the dental and medical schools. PMID- 15119839 TI - The NLM Gateway: something old, something new. AB - The NLM Gateway is a single Web interface for searching multiple NLM databases. While the majority of the available databases can be accessed directly via their own Web sites, such as PubMed, there are three databases that are unique to the NLM Gateway, including the ever-expanding OLDMEDLINE files. Thus, the Gateway is a necessary tool for librarians and researchers to be familiar with and have handy. PMID- 15119840 TI - Resource navigation for clinical authors. AB - The Web continues to grow in a largely unregulated manner. It is an established role of health sciences librarians to assist busy clinicians and other health care professionals cope with "information overload" by helping them locate useful and valuable information resources. One time-honored means of doing so is to prepare annotated bibliographies of such resources, perhaps no more so than to manage such cross-disciplinary topics as writing for publication. This article recommends resources to assist clinician writers publishing their research and other writings and suggests search strategies to update these resources as needed. PMID- 15119841 TI - The hospital library and the enterprise portal. AB - At Exempla Healthcare, the medical librarians and the e-Business staff are creating an enterprise information portal where medical reference is targeted, easily accessible, and supported by the medical librarians. A team approach has been essential. The e-Business department has worked for nine months coordinating technical challenges required to support personalization, targeted communications, and a single access point for clinical patient data. Exempla medical librarians have been involved in the definition and design of information access needs from the very beginning. The Clinicians Portal was the first developed, with other customizations to follow. Many challenges remain, but by definition, a portal is designed to be flexible and adapt to the changing needs of the enterprise it supports. PMID- 15119842 TI - Comparing the self-described searching knowledge of first-year medical and dental students before and after a MEDLINE class. AB - Do students need MEDLINE instruction? Do self-described knowledge and search skills match actual ability? To address these questions, librarians developed an assessment tool, a self-administered pre-test/post-test of eighteen skills and concepts taught in the MEDLINE class. By their own assessment, 70% or more of the medical and dental students learned something about each concept taught during the workshop. The study convinced faculty and students that the MEDLINE class is necessary. Librarians and associated faculty agree that the class is important and should continue to be mandatory. PMID- 15119843 TI - Endothelial barrier dysfunction caused by LPS correlates with phosphorylation of HSP27 in vivo. AB - Lung edema during sepsis is triggered by formation of gaps between endothelial cells followed by macrophage infiltration. Endothelial gap formation has been proposed to involve changes in the structure of the actin filament cytoskeleton. Heat shock protein 27 (HSP27) is believed to modulate actin filament dynamics or structure, in a manner dependent on its phosphorylation status. We hypothesized that HSP27 may play a role in endothelial gap formation, by affecting actin dependent events in endothelial cells. As there has been no report concerning HSP27 in lung edema in vivo, we examined induction and phosphorylation of HSP27 in lung following LPS injection, as a model of sepsis. In lung, HSP27 mainly localized in capillary endothelial cells of the alveolus, and in smooth muscle cells of pulmonary arteries. HSP27 became significantly more phosphorylated at 3 h after LPS treatment, while the distribution of HSP27 remained unchanged. Pre treatment with anti-TNFalpha antibody, which has been shown to reduce lung injury, blocked increases in HSP27 phosphorylation at 3 h. HSP27 phosphorylation was also increased in cultured rat pulmonary arterial endothelial cells (RPAEC) by treatment with TNFalpha, LPS, or H2O2. This phosphorylation was blocked by pre treatment with SB203580, an inhibitor of the upstream kinase, p38 MAP kinase. Increased endothelial permeability caused by H2O2 in vitro was also blocked by SB203580. The amount of actin associated with HSP27 was reduced after treatment with LPS, or H2O2. In summary, HSP27 phosphorylation temporally correlated with LPS induced pathological endothelial cell gap formation in vivo and in a cell culture model system. This is the first report of increased HSP27 phosphorylation associated with pathological lung injury in an animal model of sepsis. PMID- 15119844 TI - Exposure of cardiac fibroblasts to the herbicide nitrofen causes altered interactions with the extracellular matrix. AB - A large proportion of congenital heart defects result from dysmorphogenesis of valvuloseptal precursors, the endocardial cushions. Intrinsic to formation and maturation of these tissues are developmental changes in cell-cell and cell extracellular matrix interactions. Interactions between cells and the extracellular matrix play critical roles in modulating cellular processes including proliferation, migration, differentiation and even survival. While significant progress is being made in the elucidation of the cellular events involved in valvuloseptal development, little is known regarding how environmental factors may affect this process. Embryonic exposure to the herbicide nitrofen has been shown to result in congenital heart defects associated with altered endocardial cushion formation or maturation. The present studies were performed to begin to address the cellular mechanisms of these nitrofen-induced effects. Heart fibroblasts were isolated and treated with varying doses of nitrofen in vitro. Experiments were performed to determine the effects of this herbicide on important cellular processes including migration, proliferation and apoptosis. These studies illustrated a dose-dependent decrease in collagen gel contraction and proliferation in response to nitrofen. Assays were also performed to determine the effects of nitrofen on fibroblast gene expression. Increased expression of collagen type I and specific integrins were seen following nitrofen exposure. These studies illustrate that nitrofen has direct effects on cardiac fibroblast proliferation and extracellular matrix remodeling, cellular events important in valvuloseptal development. PMID- 15119845 TI - Delivery of electric pulses for DNA electrotransfer to mouse muscle does not induce the expression of stress related genes. AB - In vivo gene transfer to skeletal muscle is a promising strategy for the treatment of muscle disorders and for the systemic delivery of therapeutic proteins. Electrotransfer is a powerful method for DNA transfer into skeletal muscle. In view of the broad potential gene therapy clinical application of electrotransfer offers, it is important to perform toxicology studies on electrotransfered muscle tissue. We have investigated if the delivery of square wave electric pulses of low field strength and long duration to mouse tibial cranial muscle induced the expression of stress related genes. We have profiled gene expression patterns in muscles at different times after delivery of electric pulses using Stress/Toxicology microarrays. No significant variation in the expression of stress related-genes was detected between treated and non-treated muscles. This suggests that application of adequate, fine-tuned, electric pulses to the skeletal muscle is a non-toxic technique for gene therapy. PMID- 15119846 TI - An investigation of effects of toluene and cigarette smoking on some blood parameters and lymphocyte life span. AB - As toluene is an organic solvent, its cytotoxic effect on the cell is known. Similarly, it has been demonstrated that many of the chemical agents that enter the body through smoking have cytotoxic and genotoxic effects on the cells. In this study, the effects of these two toxic agents, both separately and in combination, on leukocyte counts, lymphocyte counts and mitotic index values were investigated. The study was carried out on blood samples of 100 males, divided into four groups: 25 non-smokers and 25 smokers, 25 toluene-exposed non-smokers and 25 no toluene-exposed smokers. The blood cell values of the blood samples were determined automatically on the hemogram apparatus. In addition slides of the blood samples were prepared according to the chromosome analysis procedure and the mitotic index values were determined through microscopy. The possible effects of smoking and toluene on lymphocyte life span was considered by correlating mitotic index values with lymphocyte counts in the same way for each of the subgroups. Results revealed that leukocyte counts and mitotic index values were higher in the smokers than the non-smokers whether or not they had been exposed to toluene. In addition the results indicate that lymphocyte life span may be shortened due to cigarette smoking and toluene exposure. PMID- 15119847 TI - The effect of enrofloxacin on cell proliferation and proteoglycans in horse tendon cells. AB - Fluoroquinolone antibiotics have been used widely in humans and domestic animals, including horses, because of their broad-spectrum bactericidal activity, and relative safety. The use of fluoroquinolones, however, is not without risk. Tendonitis and spontaneous tendon rupture have been reported in people during or following therapy with fluoroquinolones. We have studied the effects of enrofloxacin, a fluoroquinolone antibiotic used commonly in domestic animals, on tendon cell cultures established from equine superficial digital flexor tendons. Effects on cell proliferation and morphology were studied using cell counting and scanning electron microscopy. Monosaccharide content and composition was determined by gas chromatography-mass spectrometry analysis. Western and Northern blot analyses were utilized to evaluate the synthesis and expression of two proteoglycans, biglycan and decorin. Our data demonstrate that enrofloxacin inhibits cell proliferation, induces morphological changes, decreases total monosacharide content and alters small proteoglycan synthesis at the glycosylation level in equine tendon cell cultures. These effects are more pronounced in juvenile tendon cells than in adult equine tendon cells. We hypothesize that morphological changes and inhibition of cell proliferation are a result of impaired production of biglycan and decorin, proteoglycans involved in fibrillogenesis of collagen, the most important structural component of the tendon of enrofloxacin-treated tendon cells. Our findings suggest that fluoroquinolones should be used with caution in horses, especially in foals. PMID- 15119848 TI - Vitamin E protects porcine but not bovine cultured aortic endothelial cells from oxygen-derived free radical injury due to hydrogen peroxide. AB - The antioxidative effects of vitamin E (VE) are well known and have been demonstrated in in vitro studies. Since we previously observed that dextran sulfate was markedly more protective of porcine versus bovine aortic endothelial cells when damaged by hydrogen peroxide (H2O2), our objectives were to determine if a similar species difference could be observed with VE. The effects of VE or Trolox (a more water-soluble VE) against oxygen-derived free radical (OFR) injury produced by H2O2 was studied in porcine aortic endothelium (PAE) vs. bovine aortic endothelium (BAE) and bovine brain microvessel endothelium (BBME). VE or Trolox was added to culture medium for at least 24 h prior or immediately prior to H2O2 addition. In PAE, pretreatment with VE dissolved in either ethanol (VE EtOH) or Tween 20 (VE-Tween 20), or Trolox dissolved in DMSO (Trolox-DMSO) was protective, shown by increased percent viable cells and reduced lactate dehydrogenase (LDH) release. EtOH, Tween 20 or DMSO alone was protective in PAE although DMSO or Tween 20 alone was less effective than when added with VE. VE Tween 20 or Trolox-DMSO protected PAE when added just prior to H2O2 injury, but protection was significantly less than with pretreatment. DMSO immediately prior to H2O2 injury had no protective effect. Tween 20 immediately prior resulted in complete cell death. In BAE and BBME, pretreatment with VE-EtOH, EtOH, Trolox DMSO, or DMSO alone had little or no protective effect. Pretreatment with VE Tween 20 or Tween-20 alone was protective of BAE with Tween 20 being more effective than VE-Tween 20 suggesting that Tween 20 was the protective agent. These studies show that the protective effects of VE and Trolox as well as DMSO, EtOH, and Tween-20 are species dependent. PMID- 15119849 TI - Anti-Candida and mode of action of two newly synthesized polymers: a modified poly (methylmethacrylate-co-vinylbenzoylchloride) and a modified linear poly (chloroethylvinylether-co-vinylbenzoylchloride) with special reference to Candida albicans and Candida tropicalis. AB - Polymeric antimicrobial agents represent a new and important direction that is developing in the field of antimicrobial agents. Antimicrobial activity of two newly synthesized polymers: a modified poly (methylmethacrylate-co vinylbenzoylchloride) and a modified linear poly (chloroethylvinylether-co vinylbenzoylchloride) have been investigated and found to be active. Both polymers have showed a broad antimicrobial activity against C. albicans and C. tropicalis. Minimal inhibitory concentrations (MIC's) for poly (methylmethacrylate-co-vinylbenzoyl chloride) were 100, 75 and 100 microg/ml in case of C. albicans (ATCC 2091), C. albicans (SC5314) and C. tropicalis, respectively. However, polycholoroethylvinylether-covinylbenzoylchloride inhibited C. albicans (ATCC 2091), C. albicans (SC5314) and C. tropicalis with minimum inhibitory concentration values (MIC's) of 150 microg/ml against the three tested Candida strains. Mode of action studies of both polymers on the medically important yeasts, C. albicans and C. tropicalis revealed that poly (methylmethacrylate-co-vinylbenzoylchloride) induced cytotoxicity, DNA damage, and altered cell permeability and morphology, which was manifested as aggregated and swollen yeast cells (C. albicans ATCC 2091) by fluorescent microscopy examination. Poly (chloroethylvinylether-co-vinylbenzoylchloride) increased cell permeability, and respiration for C. albicans and C. tropicalis. The tested polymers at 50 microg/ml had pronounced effects on C. albicans and C. tropicalis cell wall phosphopeptidomannane, proteins, sugars and phosphorus. Generally, the two polymers proved effective against the tested microorganisms, but growth inhibitory effect varied according to the composition of the polymer active group. Many investigators consider polymeric antimicrobial agents as a potential new approach for enhancing the efficiency of some existing antimicrobial agents, including prolonged activity, reduce their toxicity, as well as reduce the environmental issues associated with product use. PMID- 15119850 TI - Nocardia beijingensis, is a pathogenic bacterium to humans: the first infectious cases in Thailand and Japan. AB - Nocardia beijingensis, a recently established new species, is an isolate from soil in China. During our taxonomic studies on 450 nocardial clinical isolates in Thailand and Japan, 17 strains from Thailand and 1 strain from Japan were found to have a similar physiological characteristic to those of N. beijingensis, such as a drug susceptibility pattern to three antimicrobial agents. Our phylogenetic studies on these 18 strains by 16S rRNA gene sequence analysis confirmed that these strains belong to N. beijingensis species. Phylogenetically, these newly isolated N. beijingensis strains were found to be classified into two distinct clades: one is a Japanese clade and other is a Chinese clade, including a reference strain and 17 Thai strains. This is the first report of human infection due to N. beijingensis strains, and we propose that the bacterium be categorized as an opportunistic infectious group regardless of its original isolation from soil. PMID- 15119851 TI - Fluconazole and itraconazole susceptibility of vaginal yeast isolates from Slovakia. AB - Vulvovaginal candidiasis is a common mucosal infection caused by opportunistic yeasts of the Candida genus. In this study, we isolated and identified the yeast species in the vagina of patients treated in the gynecology clinic and tested in vitro activities of fluconazole and itraconazole against 227 clinical yeast isolates by the NCCLS microdilution method. C. albicans (87.6%) was the most frequently identified species followed by C. glabrata (6.2%) and C. krusei (2.2%). Almost thirteen percent of yeast strains were resistant to fluconazole and 18.5% were resistant to itraconazole. Cross-resistance analyses of C. albicans isolates revealed that fluconazole resistance and itraconazole resistance were also associated with decreased susceptibilities to other azole derivatives mainly to ketoconazole and miconazole. At the same time no cross resistance to polyene antibiotics amphotericin B and nystatin was observed. These results support the notion that antifungal agents used to treat vaginitis may be contributing to the drug resistance problem by promoting cross-resistance to a range of clinically used antifungals. PMID- 15119853 TI - Observation of protoplasts from the conidia of Arthrinium aureum by scanning electron microscopy. AB - Protoplast formation from conidia of Arthrinium aureum was achieved with Lysing Enzyme L-2265 (Sigma Chemical) from Trichoderma harzianum. Scanning electron microscopy of conidia protoplasts showed the acquired spherical shape. PMID- 15119852 TI - Cladosporium and respiratory allergy: diagnostic implications in Saudi Arabia. AB - An allergological study to evaluate allergenicity to Cladosporium, Burkard 7-Day Volumetric Spore Trap and Personal Volumetric air sampler (viable mode) were employed to conduct air sampling for 12 months in three regions of Saudi Arabia. The study was extended for a continuous 3rd year at one site. Skin prick testing (SPT) was also conducted on 605 allergic individuals using commercial extracts of C. herbarum. Cladosporium emerged to be the most prevalent genus in the outdoor environment constituting up to 25% of all fungal spores in the dry region and 37.1 and 41.2% in two coastal cities respectively. Amongst the species C. sphaerospermum, C. macrocarpum, C. cladosporioides and C. herbarum were noted. Maximum hourly concentrations up to 14 x 10(3) m(-3) were recorded in coastal region during winter months. Morning concentrations were higher at both city sites compared to afternoon concentration. SPT result revealed an overall 19.67% positive reactions with majority showing mild reactions. PMID- 15119854 TI - Presence of vacuoles in Arthrinium aureum. PMID- 15119855 TI - Antimicrobial activities of selected Cyathus species. AB - Twelve selected Cyathus species were tested for their abilities to produce antimicrobial metabolites. Most of them were found to produce secondary exo metabolites that could induce morphological abnormalities of rice pathogenic fungi Pyricularia oryzae. Some extracts from the cultivated liquid obviously inhibited human pathogenic fungi Aspergillus fumigatus, Candida albicans and Cryptococcus neoformans. Activities against six human pathogenic bacteria were also obtained from some of these extracts. PMID- 15119856 TI - Effects of nematicides on cotton root mycobiota. AB - Baseline information on the diversity and population densities of fungi collected from soil debris and cotton (Gossypium hirsutum L.) roots was determined. Samples were collected from Tifton, GA, and Starkville, MS containing cotton field soil treated with the nematicides 1,3-dichloroproprene (fumigant) and aldicarb (granules). A total of 10,550 and 13,450 fungal isolates were collected from these two study sites, respectively. Of this total, 34 genera of plant pathogenic or saprophytic species were identified. Pathogenic root fungi included Fusarium spp. (40% of all isolations), Macrophomina, Pythium, Rhizoctonia, and Sclerotium. Fusarium and Rhizoctonia were the most common fungal species identified and included F. oxysporum, F. verticillioides and F. solani, the three Fusarium species pathogenic on cotton plants. Population densities of Fusarium were not significantly different among locations or tissue types sampled. Macrophomina was isolated at greater numbers near the end of the growing seasons. Anastomosis groups of R. solani isolated from roots and soil debris included AG-3, -4, -7, 2 2, and -13 and anastomosis groups of binucleate Rhizoctonia included CAG-2, -3, and -5. Occurrences and frequency of isolations among sampling dates were not consistent. Fluctuations in the frequency of isolation of Rhizoctonia did not correspond with changes in frequency of isolation of the biological control fungus, Trichoderma. When individual or pooled frequencies of the mycobiota were compared to nematicide treatments, no specific trends occurred between treatments, application methods or rates. Results from this study show that use of 1,3-D and aldicarb in cotton fields does not significantly impact plant pathogenic fungi or saprophytic fungal populations. Thus cotton producers need not adjust seedling disease control measures when these two nematicides are used. PMID- 15119857 TI - Enzyme production by the mycoparasite Verticillium biguttatum against Rhizoctonia solani. AB - Verticillium biguttatum, a mycoparasite of the ubiquitous soil-borne plant pathogen Rhizoctonia solani, excreted chitinase and beta-1,3-glucanase into liquid medium when grown on laminarin and chitin, respectively. Neither chitinase nor beta-1,3-glucanase was produced by the mycoparasite when grown on cell walls of two isolates of R. solani representing anastomosis groups (AG)-3 and AG-8. Extracellular protease was induced by growth on cell walls of the pathogen, whereas beta-1,3-glucanase and chitinase were produced bound to the cell wall of V. biguttatum. This is the first report of chitinase, beta-1,3-glucanase and protease production by V. biguttatum. These enzymes may play a previously unforeseen role in dissolving and penetrating the cell walls of R. solani. PMID- 15119858 TI - Pod and seed mycoflora on transgenic and conventional soybean [Glycine max (L.) Merrill] cultivars in Mississippi. AB - A 2-year (1999-2000) study was conducted at Starkville and Stoneville, MS to determine if the occurrence of the mycoflora varied on Roundup Ready (transgenic) compared to conventional soybean (Glycine max) cultivars. A total of 7,658 fungal isolates were identified from the pod and seed tissues of four cultivars compared at growth stages R6 and R8. Ninety-nine percent of all fungi isolated were mitosporic fungi and ascomycetes. In both years, total fungal isolates from the two locations were greater from the pod (65%) than from seed (33%) tissues. Isolation frequency from conventional cultivars was 54% compared to 46% for the transgenic cultivars. The most common fungi identified that are reported pathogens of soybean included Alternaria, Cercospora, Cladosporium, Diaporthe, Fusarium and Verticillium spp. When main effects and interactions were compared among the frequency data for the fungal genera, significant differences occurred, but consistent trends were not noted. Isolation frequencies of Diaporthe spp. during the R6 growth stage, were significantly greater on the conventional than on the transgenic cultivars in both years of the study, but only at Starkville. Isolation frequencies from samples taken during the R8 growth stage were similar at both locations in 1999 and 2000. Fusarium spp. isolated at R6 and R8 growth stages from pod and seed tissues were significantly greater on conventional than on transgenic cultivars in 2000. Even though frequencies were often significantly different between the transgenic and conventional cultivars, the data was not consistent between locations, pod and seed tissues, or growth stages. The pod and seed mycoflora of transgenic and conventional soybean cultivars was, therefore, similar in Mississippi. PMID- 15119859 TI - Penicillium implicatum causes a destructive rot of pomegranate fruits. AB - In this study Penicillium implicatum Biourge was found to be the cause of a destructive rot of stored pomegranate (Punica granata) fruits. This species has not previously been reported to cause disease of the fruit. The ability of the strain to decay pomegranates as well as some varieties of apples (Jonagold, Selena and Vanda) under laboratory conditions after inoculation with conidia into the subcutaneous layer of healthy fruits was demonstrated. The fungus was also found on mouldy stamens of pomegranates. PMID- 15119860 TI - Influence of temperature on Pythium splendens--induced root disease on carambola, Averrhoa carambola. AB - A series of glasshouse and incubator studies were conducted to investigate the role played by Pythium splendens in a decline disorder of carambola, Averrhoa carambola. Plants, 4-6 months old, were grown in native calcareous soil either infested or not infested with the pathogen. Isolates recovered from atemoya, carambola and passion fruit grew optimally at 30 degrees C, and significantly (P < 0.05) increased root necrosis and reduced root, shoot and total biomass of carambola. Temperature had a profound impact on the latter relationships. Two or more times more necrosis developed at 10 and 15 degrees C than at 25 and 30 degrees C. Total biomass accumulations were over four times greater at 30 degrees C than at 10 degrees C, and were always lower in soil infested with P. splendens. When biomass totals from infested and noninfested soil were compared, relative values were lowest at 15 and 20 degrees C and were almost two times greater at 30 degrees C than at 20 degrees C. Root infection by P. splendens was greatest at 15 and 20 degrees C, far below the species' optimum for growth, and at 30 degrees C was over nine times lower than at 15 and 20 degrees C. This is the first detailed report of P. splendens as a pathogen of carambola. PMID- 15119861 TI - Aflatoxin binders II: reduction of aflatoxin M1 in milk by sequestering agents of cows consuming aflatoxin in feed. AB - Sequestering agents bind dietary aflatoxin B1 (AFB1) and reduce absorption from an animal's gastrointestinal tract. As a result, they protect an animal from the toxic effects of AFB1 and reduce transfer of the metabolite, aflatoxin M1 (AFM1), into milk. Three experiments, using late-lactation Holstein cows fed AFB1 contaminated feed, were conducted to evaluate several potential sequestering agents for their abilities to prevent or reduce the transmission of AFM1 into milk. Six agents previously tested in our laboratory for AFB1 binding in vitro were evaluated in these experiments. These were: SA-20, an activated carbon (AC A); Astra-Ben-20, a sodium bentonite (AB-20); MTB-100, an esterified glucomannan (MTB-100); Red Crown, a calcium bentonite (RC); Flow Guard, a sodium bentonite (FG); and Mycrosorb, a sodium bentonite (MS). Five of the six sequestering agents significantly (P < 0.01) reduced AFM1 contamination of milk (AB-20, 61%; FG, 65%; MS, 50%; MTB-100, 59%; and RC, 31%); whereas, AC-A, activated carbon, had no effect on AFM1 transmission at 0.25% of feed. By the first milking (1 day after cows consumed contaminated feed), AFM1 appeared in milk, then reached maximum levels after three days, and was absent from milk within four days after AFB1 was removed from the feed. Sodium bentonites at 1.2% of feed showed good potential as AFB1 binders; MTB-100, a yeast cell wall product, was equally effective at 0.05% in feed. Potential AFB1 binding agents should be evaluated experimentally to demonstrate efficacy. Our data show that sequestering agents can reduce AFM1 in milk of cows fed AFB1-contaminated feed. PMID- 15119862 TI - Biomechanical aspects of prosthetic treatment of structurally compromised teeth. AB - PURPOSE: This article presents clinical guidelines for restoring structurally compromised teeth and dentitions to reduce the risk for fatigue-caused failures in connection with prosthetic reconstructions. MATERIALS AND METHODS: Based on the best scientific evidence available and clinical expertise acquired through experience and practice, biomechanical principles are elucidated from a prosthetic aspect. RESULTS: In prosthetic treatment in the structurally compromised dentition, all efforts need to be focused on protecting the abutments and reconstruction from future fatigue failures. A modified, "therapeutic" occlusion to avoid nonaxial forces may then be prudent. CONCLUSION: By lending the prosthesis a favorable occlusal design, the nonaxial forces may be markedly reduced, and the teeth, cement, and restorative materials will be less susceptible to fatigue failures. PMID- 15119863 TI - Strength, reliability, and mode of fracture of bilayered porcelain/core ceramics. AB - PURPOSE: The aim of this investigation was to compare the biaxial flexural strength, its reliability, and the mode of fracture of bilayered disks made of two core materials (In-Ceram Alumina and In-Ceram Zirconia), both veneered with conventional feldspathic porcelain (Vita Alpha). MATERIALS AND METHODS: One hundred forty specimens (monolithic and bilayered) of In-Ceram Alumina, In-Ceram Zirconia, and Vita Alpha were made and tested with the biaxial flexural test. Finite element analysis was used to estimate the maximum tensile stress at fracture. Data were analyzed with one-way ANOVA, Tukey HSD, and Weibull distribution. SEM was used to identify the initial crack and characterize the fracture mode. RESULTS: All specimens with the core material on the bottom surface were statistically significantly stronger and more reliable than those with the porcelain on the bottom surface. Among them, In-Ceram Zirconia was stronger than In-Ceram Alumina. There was no statistically significant difference among groups when the porcelain underwent tension. Two different modes of fracture were observed in the bilayered samples according to which material was on the bottom surface. CONCLUSION: The material that underwent tensile stress dictated the strength, reliability, and fracture mode of the specimens. The design of the restorations and the actual distribution of the tensile stresses must be taken into account; otherwise, the significant contribution of stronger and tougher core materials to the performance of all-ceramic restorations may be offset by the weaker veneering porcelain. PMID- 15119864 TI - Assessment of dental appearance following changes in incisor angulation. AB - PURPOSE: This study evaluated the assessment of attractiveness of standardized changes in incisor angulation of different tooth arrangements. MATERIALS AND METHODS: Four sets of images showing the maxillary anterior teeth without lips against a black background were used for evaluation. Each set contained the original photograph, one computer-manipulated symmetric image, and four images with different standardized changes in incisor angulation (10 degrees). The judges, consisting of three groups (30 dental students, 30 medical students, 30 art students), ranked each photo set for attractiveness from 1 (most attractive) to 6 (most unattractive). RESULTS: The mean ranking (in parentheses) showed that symmetric teeth with ideal axes (2.5) and changes in the angulation of one (2.8) or both (2.5) lateral incisors were significantly more attractive than the angulation of one (4.2) or both (4.6) central incisors. There were no differences between the groups of participants. All findings were reproducible. CONCLUSION: Tooth arrangements showing central incisors with ideal axes were more attractive. Slight changes in the angulation of one or both lateral incisors did not influence attractiveness negatively. PMID- 15119865 TI - An introduction to silanes and their clinical applications in dentistry. AB - PURPOSE: This overview presents a description of organofunctional trialkoxysilane coupling agents (silanes), their chemistry, properties, use, and some of the main clinical experiences in dentistry. MATERIALS AND METHODS: The main emphasis was on major dental journals that have been reviewed from 1958 up to the latest research news from 2002. A MEDLINE search with the key words "dental silanes" was used. Special silane literature and journals outside dentistry were also cited. RESULTS: The main emphasis is on the use of silanes in prosthetic and restorative dentistry. Clinical relevance was based mainly on either short- or long-term tests. The interpretation of various results is not given, mainly because of controversial observations that may be very difficult to explain. Nevertheless, the majority of the clinical results pointed to silanes playing a significant role in the adhesion process. Silane reaction mechanisms were not entirely understood, and there exist several theories for bonding mechanisms for silanes and substrates. CONCLUSION: Dental materials offer a continuously challenging forum for silanes, and silanes will play an essential role in material development. PMID- 15119866 TI - A survey of disinfection of irreversible hydrocolloid and silicone impressions in European Union dental schools: epidemiologic study. AB - PURPOSE: The objective of this cross-sectional study was to describe corresponding procedures for irreversible hydrocolloid and silicone impressions taught and used in European Union dental schools. MATERIALS AND METHODS: A self administered questionnaire requesting information about rinsing and disinfection methods was sent to each of the 373 heads of prosthodontic, pedodontic, and orthodontic departments in the 131 European Union dental schools. Response rate was 94%. Statistical analysis included chi-square or Fisher exact tests, and ANOVA or Kruskall-Wallis tests. RESULTS: Of the responding departments, 92% systematically rinsed their impressions. Fifteen percent of the departments, mostly orthodontics, never disinfected irreversible hydrocolloid impressions, and 11% never disinfected silicone impressions. The immersion method was used by 65% for irreversible hydrocolloid impressions (73% for silicone), with a disinfection time of 10.3 +/- 6.3 minutes (11.8 +/- 7.4 for silicone). The disinfected impressions were not rinsed by 16% for irreversible hydrocolloid and 14% for silicone. Most departments used brand-name products. CONCLUSION: The same disinfection procedure for both irreversible hydrocolloid and silicone impressions was used by 78% of departments. There was great diversity, however, between departments in the procedure used for each impression material. PMID- 15119867 TI - Emotional effects of tooth loss in community-dwelling elderly people in Hong Kong. AB - PURPOSE: This study investigated the emotional effects of tooth loss among community-dwelling elderly people in Hong Kong and compared the effects among edentulous and partially dentate elderly. MATERIALS AND METHODS: A questionnaire study involved 233 southern Chinese elderly who were recruited and interviewed at social centers for elderly people throughout Hong Kong. Data were analyzed using chi-square or chi-square exact tests. RESULTS: Twenty-two percent of the participants reported difficulty accepting tooth loss, with no difference between the edentulous and partially dentate. However, 95% stated that their confidence was unaffected. For more than half of elderly people, tooth loss had a negative effect on food choice and enjoyment of food, with removable denture wearers having greater restrictions. Avoiding going out, eating in public, and forming close relationships were less-common problems. Twenty-two percent felt unprepared for the effects of tooth loss, and more than half said that better communication with the dentist would have helped. CONCLUSION: The emotional effects of tooth loss were not marked among elderly people, and there were no differences between edentulous and partially dentate individuals. However, significant disability was experienced because of restrictions in daily living activities. Restrictions were more severe in people who had lost enough teeth to necessitate denture wearing. PMID- 15119868 TI - Stability of the screw joints in patients with implant-supported fixed prostheses in edentulous jaws: a 1-year follow-up study. AB - PURPOSE: The purpose of this follow-up study was to evaluate the stability of the screw joint in edentulous patients 1 year after treatment with implant-supported fixed prostheses (Branemark system). MATERIALS AND METHODS: A total of 20 patients were included, 10 treated in the maxilla and 10 in the mandible. The fixed prostheses were removed approximately 1 year after insertion, and the stability of the screw joints was evaluated using a rating scale based upon the CDA quality evaluation criteria of dental care. RESULTS: All implant-supported fixed prostheses were recorded as stable before the prosthetic screws (gold screws) were unscrewed. "Unacceptable loosening" was observed in 4% of the prosthetic screws and in 29% of the abutment screws. CONCLUSION: In this study, only a few of the prosthetic screws showed unacceptable loosening after 1 year of function. The clinical relevance of the observed high occurrence of loose abutment screws could be questioned, as all fixed prostheses were initially recorded as stable. PMID- 15119869 TI - A cost comparison of mandibular two-implant overdenture and conventional denture treatment. AB - PURPOSE: This article compares the cost of mandibular two-implant overdenture treatment to that of conventional denture treatment in an academic teaching hospital. MATERIALS AND METHODS: Sixty edentulous patients (aged 65 to 75 years) participated in a randomized clinical trial. All patients received a new maxillary complete denture and either a mandibular conventional denture (n = 30) or an implant overdenture on two unsplinted implants (n = 30). Resource-based microcosting of direct and indirect costs (eg, expenses and time cost to patients) of all scheduled and unscheduled visits was conducted through 1 year following delivery of the prostheses. RESULTS: Mean direct costs (1999 CD dollars) for scheduled visits in the implant and conventional groups were 2,332 dollars and 814 dollars, respectively, and mean indirect costs were 1,150 dollars and 810 dollars, respectively. Differences between the two groups were significant. Twenty-six patients in each group had unscheduled visits during the study at a median direct cost for the overdentures of 85 dollars and 64 dollars for the conventional dentures. Median indirect costs for unscheduled visits were 163 dollars and 202 dollars, respectively. These differences were not significant. Mean total costs of the overdentures were 4,245 dollars and 2,316 dollars for the conventional dentures, and the between-group difference was significant. CONCLUSION: The direct cost of mandibular two-implant overdenture treatment was 2.4 times higher than that of conventional denture treatment. When indirect costs were added, the implant-to-conventional total cost ratio estimate was 1.8. These cost data can now be combined with estimates of the efficacy of the two types of prosthesis so practitioners and patients can make informed decisions about these prosthodontic treatment concepts. PMID- 15119870 TI - Retrospective evaluation of the success of oral rehabilitation using the Frialit 2 implant system. Part 1: Influence of topographic and surgical parameters. AB - PURPOSE: This retrospective longitudinal study evaluated the success of implant prosthetic rehabilitation with the Frialit-2 implant system. MATERIALS AND METHODS: The study was performed with 504 patients, from two treatment centers, who had received a total of 1,554 implants between May 1990 and May 2001. The data of these patients, who received the implants in various regions and for various indications, were analyzed with respect to clinical, topographic, and radiographic parameters. The mean observation period was 6.2 years, with a maximum of 134 months. RESULTS: A survival rate of 94.8% was found for all implants. The implant survival rate of 92.6% in the maxilla remained constant after 68 months of observation. In the mandible, the implant survival rate of 96.7% showed no changes after 76 months. Kaplan-Meier analysis identified jaw, occurrence of postoperative complications, and region as statistically significant factors influencing implant survival. Multivariate Cox regression showed that gender, occurrence of postoperative complications, and jaw were factors that increased the risk of implant loss. Statistically significant correlations were found between the incidence of implant loss and vertical bone loss adjacent to the implant at the time of second-stage surgery. CONCLUSION: Implant survival rate is influenced by implant site, gender, and occurrence of complications. On the whole, the Frialit-2 system proved successful in all areas of indication after long-term observation. PMID- 15119871 TI - Water sorption and dimensional stability of three glass fiber-reinforced composites. AB - PURPOSE: Water sorption and dimensional stability of three fiber-reinforced composites were studied. Two composites (Vectris, FibreKor) were resin impregnated industrially, and one composite (Stick) was polymer preimpregnated but required further manual impregnation. MATERIALS AND METHODS: Bar-shaped specimens of each material were prepared according to manufacturers' instructions. The water sorption and dimensional change of each specimen were calculated according to the change in its weight and dimension before and after immersion. Specimens were immersed in distilled water for 1, 7, 60, and 180 days. SEMs were taken to examine the quality of the fiber-matrix interface. The volume percentage of fiber content of each fiber-reinforced composite was experimentally estimated. RESULTS: A general trend of increasing water sorption for each immersion period according to the material type was: Vectris < FibreKor < Stick. There were no significant differences in dimensional change among the materials and immersion periods. CONCLUSION: The preimpregnated fiber-reinforced composite (Stick) showed higher water sorption than the industrially impregnated fiber reinforced composites (Vectris, FibreKor). Despite a variation in the water sorption of the fiber-reinforced composites studied, all were within a 32 microg/mm3 criterion established by the ISO. The magnitude of dimensional change was small enough that it should not raise any significant clinical concern. PMID- 15119872 TI - Temperature increase during resin cement polymerization under a ceramic restoration: effect of type of curing unit. AB - PURPOSE: This study measured the temperature increase induced by various types of curing units during resin cement polymerization under ceramic restorations. MATERIALS AND METHODS: The resin cement was polymerized between a ceramic specimen (diameter 5 mm, height 2 mm) and a dentin disk (diameter 5 mm, height 1 mm) with a conventional halogen light, a high-intensity halogen light, a plasma arc light, and a light-emitting diode unit. The temperature increase was measured under the dentin disk with a J-type thermocouple wire connected to a data logger. Ten measurements were carried out for each curing unit. Difference between starting and highest temperature readings was measured, and the 10 calculated temperature changes were averaged. RESULTS: Temperature increase varied significantly depending on curing unit used. The plasma-arc light induced significantly higher temperature increases than any other curing unit. The light emitting diode unit produced the lowest temperature changes. There were no statistically significant differences between the conventional and high-intensity halogen curing units. CONCLUSION: Polymerization with curing units characterized by high energy output (plasma-arc light) caused higher temperature changes compared to other curing units, but the temperature increase detected was not viewed as critical for pulpal health. PMID- 15119873 TI - Incisal tooth wear and self-reported TMD pain in children and adolescents. AB - PURPOSE: Incisal tooth wear may be a sign of long-term bruxing behavior. Bruxism is purported to be a risk factor for temporomandibular disorders (TMD). The aim of this population-based cross-sectional study was to determine if anterior tooth wear is associated with the self-report of TMD pain in children and adolescents. MATERIALS AND METHODS: In a population sample of 1,011 children and adolescents (mean age 13.1 years, range 10 to 18 years; female 52%; response rate 85%), TMD cases were defined as subjects reporting pain in the face, jaw muscles, and temporomandibular joint during the last month according to RDC/TMD. All other subjects were considered controls. Incisal tooth wear was assessed in the clinical examination using a 0 to 2 scale (no wear, enamel wear, dentin wear) for every anterior permanent tooth. The mean wear score for the individuals was categorized into 0, 0.01 to 0.20, 0.21 to 0.40, and 0.41+. A multiple logistic regression analysis, controlling for the effects of age and gender, analyzed the association between the categorized summary wear score and TMD. Specifically, the hypothesis of a trend between higher tooth wear scores and higher risk of TMD was tested. RESULTS: An odds ratio of 1.1 indicated, after adjusting for gender and age, no statistically significantly higher risk of TMD pain with higher tooth wear scores. CONCLUSION: Incisal tooth wear was not associated with self-reported TMD pain in 10- to 18-year-old subjects. PMID- 15119874 TI - A survey of the use of mandibular implant overdentures in 10 countries. AB - PURPOSE: This preliminary international survey compared provision of implant retained overdentures to fixed implant-supported prostheses for edentulous mandibles. MATERIALS AND METHODS: Questionnaires based on a 2001 Swedish study were sent to prosthodontists and specialist clinics in nine additional countries. RESULTS: Response rate varied from 53% to 100% in 10 national surveys and should allow careful comparison of results. The relationship between implant overdentures and fixed implant-supported prostheses in treatment of edentulous mandibles varied much; in Sweden, the proportion of overdentures was 12%, whereas it was 93% in The Netherlands. In all countries, the most common reason for choice of the overdenture was reduced cost. In all but two countries, the majority of respondents thought that patients with implant overdentures were equally or more satisfied with overdentures as those with fixed implant-supported prostheses. CONCLUSION: There were great differences among the 10 countries in choice of implant treatment of the edentulous mandible. The relative proportion of mandibular overdentures to fixed prostheses was low in Sweden and Greece and varied from one to two thirds in the other countries, except The Netherlands. PMID- 15119875 TI - Marginal adaptation of fixed prosthodontics: a new in vitro 360-degree external examination procedure. AB - PURPOSE: This study evaluated the marginal fit of experimental and custom-made fixed prosthetic restorations through a new 360-degree external examination. The minimum number of gap measurements required to produce relevant results for gap analysis was also investigated. MATERIALS AND METHODS: The marginal fit of six experimental and eight custom-made crowns was observed microscopically by means of a mechanical device, and software was employed to measure the gap. Two crowns, chosen from among the 14 previously evaluated, were reanalyzed to determine the minimum number of gap measurements required to produce significant results for gap analysis. RESULTS: The precision obtained with the custom-made crowns differed from that of the experimental specimens. The minimum number of measurements required to produce a sample mean value within +/- 5 microm of the mean, calculated over 360 measurements, taking standard error of the means < or = 4 microm, was 18 for experimental and 90 for custom-made crowns, for both equidistant measurement spacing and randomly selected points. CONCLUSION: Differences in fit between experimental specimens and custom-made ones showed that experimental results might not always be obtained in clinical practice. Within the limitations of the protocol of this study, the minimum number of measurements required to ensure relevant results for gap analysis was 18 for experimental and 90 for custom-made crowns. PMID- 15119876 TI - Decayed/missing/filled teeth and shortened dental arches in Tanzanian adults. AB - PURPOSE: This study assessed decayed/missing/filled teeth (DMFT), presence of occlusal units, and prevalence of shortened dental arches in a Tanzanian adult population. MATERIALS AND METHODS: The dental state of samples of the Tanzanian population was studied. Oral examinations were conducted on 5,532 adults from rural and urban cluster samples. DMFT was related to age, gender, and residence. The pattern of tooth loss within dental arches was mapped, and the prevalence of shortened dental arches was estimated. RESULTS: Mean DMFT scores increased gradually from 1.8 (20- to 29-year-olds) to 3.8 (50- to 59-year-olds). A steep increase was observed in the > or = 60-year-olds (DMFT 8.1). "Missing" was the dominant component of DMFT. The mean number of present teeth ranged from 27 in the youngest to 20 in the oldest age group. Under 60 years of age, DMFT was significantly higher for women than for men. No differences were found between residence categories. Molars were more frequently decayed, missing, and mobile than premolars. Of all subjects, 41% had complete dental arches, 44% had interruptions (of these, 73% were in posterior regions only), and 15% had shortened dental arches; 0.5% were edentulous. Of the subjects with shortened dental arches, about 65% had at minimum three pairs of occluding premolars. CONCLUSION: An initial low DMFT rate increased after the age of 60 years. Molars had the highest risk of dental decay and were most frequently absent. Shortened dental arches develop as a consequence of the pattern of tooth decay and tooth loss, although interruptions were frequently seen in the posterior regions. PMID- 15119877 TI - A prospective randomized clinical study comparing implant-supported fixed prostheses and overdentures in the edentulous mandible: prosthodontic production time and costs. AB - PURPOSE: This work compared implant-supported fixed prostheses and overdentures in the edentulous mandible in a randomized prospective study of treatment results, clinical working hours, laboratory working hours, and laboratory costs including materials. MATERIALS AND METHODS: Seventeen subjects agreed to participate. Three standard Branemark implants were placed between the mental foramina in each patient. After the connection of standard abutments, subjects were randomly assigned to the fixed prosthesis group (n = 11) or overdenture group (n = 6). Subjects in the fixed group were provided with prostheses according to the All-in-One concept. Subjects in the overdenture group received a conventional denture retained by a Dolder bar system. RESULTS: One implant was lost in the fixed prosthesis group after 1 year. A new implant was placed, and the prosthesis was refitted. Mean clinical working hours were 3.1 in the fixed prosthesis group and 4.1 in the overdenture group. Mean laboratory working hours were 12.5 in the fixed prosthesis group and 7.7 in the overdenture group. Total laboratory costs, including materials, were on average about 1,700 US dollars for the fixed prosthesis and 1,350 US dollars for the overdenture. CONCLUSION: A fixed implant-supported prosthesis in the edentulous mandible could be provided at about the same cost as an overdenture using the method described. Provided that the early survival results prove to be long lasting, the choice between a fixed and a removable prosthesis need not be a matter of economy. PMID- 15119878 TI - Randomized controlled clinical trial on satisfaction with resilient denture liners among edentulous patients. AB - PURPOSE: The purpose of this study was to measure patients' satisfaction and their preference between mandibular dentures with permanent silicone-based resilient denture liner (SR) and conventional heat-activated acrylic resin (AR), both opposed by acrylic resin-based maxillary complete dentures. MATERIALS AND METHODS: Twenty-eight edentulous patients who had fulfilled selection criteria and provided informed consent were enrolled in this trial. Subjects were allocated randomly to either arm of cross-over groups (AR-SR/SR-AR), stratified by gender, using a random permuted block within the strata method. The AR-SR group received AR denture treatment followed by SR denture treatment. The SR-AR group received treatment in the reverse sequence. The primary outcome was patient satisfaction measured on 100-mm VAS, analyzed by two-way ANOVA and the Bonferroni multiple comparison as a post hoc test. The secondary outcome was patients' preference, evaluated by chi-square goodness-of-fit test. An intention-to-treat analysis was performed. RESULTS: Twenty-five subjects were enrolled in the analysis. There were no significant differences between AR and SR dentures 1, 2, and 3 months after the completion of control. Eighteen of 25 patients preferred SR dentures. CONCLUSION: Although there were no significant differences in patient satisfaction ratings between the two types of dentures, a significant majority of patients preferred those with a resilient denture liner. PMID- 15119879 TI - A 10-year prospective evaluation of CAD/CAM-manufactured (Cerec) ceramic inlays cemented with a chemically cured or dual-cured resin composite. AB - PURPOSE: The present follow-up study was carried out to evaluate the performance of Class II Cerec inlays after 10 years of clinical service. MATERIALS AND METHODS: Sixty-six Class II CAD/CAM ceramic inlays were placed in 27 patients. Each patient received at least one inlay luted with a dual-cured resin composite and one inlay luted with a chemically cured resin composite. At the 10-year recall, 25 (93%) patients with 61 (92%) inlays were available for evaluation using a slight modification of the USPHS criteria. RESULTS: Fifty-four (89%) of the 61 inlays reevaluated still functioned well at the 10-year recall. During the follow-up period, seven (11%) of the inlays required replacement because of: four inlay fractures, one cusp fracture, endodontic problems in one case, and postoperative symptoms in one case. All the replaced inlays had been luted with the dual-cured resin composite. The fractured inlays were all placed in molars. The estimated survival rate after 10 years was 89%, 77% for the dual-cured resin composite-luted inlays and 100% for the chemically cured resin composite-luted ones. The difference was statistically significant. CONCLUSION: Patient satisfaction with and acceptance of the Cerec inlays were high, and the performance after 10 years of clinical service was acceptable, especially regarding the inlays luted with the chemically cured resin composite. The properties of the luting agents seem to affect the longevity of the type of ceramic inlays evaluated. PMID- 15119880 TI - Individualized nasal mask fabrication for positive pressure ventilation using dental methods. AB - PURPOSE: The aim of this study was to develop a simple technique to manufacture individualized ventilatory nasal masks for pediatric patients using materials and procedures commonly applied in dentistry. MATERIALS AND METHODS: Three cases of pediatric patients who met with severe difficulties in their adaptation to commercially available nasal masks are described: one premature infant, one child diagnosed with achondroplasia, and one child with congenital central hypoventilation syndrome. RESULTS: In each case, a light nasal mask was designed with two independent parts that become perfectly adapted to the patient's nose: one soft for the skin contact, and another rigid for dimensional stability. In all patients, adequate levels of ventilation were reached. CONCLUSION: This easy, inexpensive nasal mask fabrication technique can be used in a great number of patients, increasing the efficacy of individualized masks. PMID- 15119881 TI - Final impressions: a review of material properties and description of a current technique. AB - Esthetic rehabilitations are characterized by a sequence of well-structured clinical and laboratory steps, during which different kinds of impressions are required. This review presents a survey of the most clinically relevant physical properties that characterize final impression materials and their interactions with the products they are commonly in contact with. The principal steps of an esthetic rehabilitation involving a diagnostic phase, together with a rational step-by-step approach to final impressions, are described. The one-step/double mix impression using polyvinyl siloxane materials associated with a "double cord" gingival displacement is explained. PMID- 15119882 TI - Treatment of chronic desquamative gingivitis using tissue-engineered human cultured gingival epithelial sheets: a case report. AB - Human cultured gingival epithelial sheets were used as an autologous grafting material for regenerating gingival tissue in the maxillary left and mandibular right quadrants of a patient with chronic desquamative gingivitis. Six months post-surgery in both treated areas, there were gains in keratinized gingiva and no signs of gingival inflammation compared to presurgery. In the maxillary left quadrant, preoperative histopathologic findings revealed the epithelium was separated from the connective tissue and inflammatory cells were extensive. After grafting with the gingival epithelial sheets, inflammatory cells were decreased and separation between epithelium and connective tissue was not observed. The human cultured gingival epithelial sheets fabricated using tissue engineering technology showed significant promise for gingival augmentation in periodontal therapy. PMID- 15119883 TI - Clinical and histologic evaluation of anorganic bovine bone collagen with or without a collagen barrier. AB - This study evaluated an anorganic bovine-derived xenograft (Bio-Oss Collagen) in the treatment of human periodontal defects. Four patients with intrabony defects on teeth that were treatment planned for extraction were enrolled in the study. Presurgical measurements of probing depth, attachment level, and recession were recorded. The surgical procedure consisted of flap reflection, debridement of the osseous defects and root surface, placement of a notch through calculus into the root surface, topical application of a tetracycline paste to the root surface, grafting with Bio-Oss Collagen, and flap closure. Three of the eight defects examined received a resorbable collagen barrier (Bio-Gide) in addition to the bone graft. Patients were seen every 2 weeks for plaque control and review of oral hygiene measures. Six months postsurgery, clinical parameters were rerecorded prior to en bloc resection of teeth and adjacent graft sites. The majority of sites showed a favorable clinical response with respect to probing depth reduction and clinical attachment gain. Histologic analysis demonstrated new bone, cementum, and periodontal ligament coronal to the reference notch in two of the eight specimens. Two sites demonstrated new attachment, and four showed a long junctional epithelium. Periodontal regeneration is possible following a bone-replacement graft of Bio-Oss Collagen. PMID- 15119884 TI - Comparative study of manual and ultrasonic instrumentation of cementum surfaces: influence of lateral pressure. AB - The goal of periodontal treatments is to eliminate bacteria and their products without damaging cementum surfaces. Nonsurgical treatments are often limited by the inability of curettes to access the most apical zone of the pocket. While ultrasonic mini-inserts have been used for nearly 10 years now, their effect on dental tissues has not been tested. The purpose of the present study was to compare a new series of mini-inserts to Gracey curettes, which are the reference in nonsurgical treatments. Two experienced periodontists conducted root treatments on teeth destined for extraction using regular clinical criteria. One face of each root was instrumented using a Gracey curette, and the opposite face was instrumented using an ultrasonic mini-insert. After the instrumentation procedure, the teeth were prepared for examination by secondary electron (topographic features) and backscattered electron (organic and mineral composition) microscopy. Differences in surface composition between teeth treated by the two periodontists were noted and were related to the lateral pressure exerted. Calculus removal was less effective when strong lateral pressure was exerted using the ultrasonic mini-inserts, while more cementum was removed and more scratching occurred with both manual and ultrasonic instruments. In all cases, the ultrasonic mini-inserts allowed greater apical access. The new ultrasonic mini-inserts were as effective as manual curettes in eliminating plaque and calculus. The shape of the mini-inserts made them more effective in apical zones. The amount of damage to the cementum depended on the lateral pressure exerted by the periodontist. PMID- 15119885 TI - A zirconia implant-crown system: a case report. AB - Exposed metal crown margins in the porcelain-fused-to-metal technique might be an esthetic problem. Therefore, all-ceramic crown systems are used for restoring teeth and dental implants. In addition, the gray color of a titanium implant might hamper the esthetic appearance of the entire reconstruction in cases of thin peri-implant soft tissue or tissue retraction. To further improve the esthetic aspect of dental implants, efforts are undertaken to develop implant systems fabricated out of tooth-colored materials that are biocompatible and able to withstand masticatory forces. One such material may be zirconia. The present article presents a case in which an all-ceramic custom-made zirconia implant crown system was used for the replacement of a single tooth. PMID- 15119886 TI - Effect of autogenous harvest site location on the outcome of ridge augmentation for implant dehiscences. AB - Treatment planning for the placement of an implant in a site with a thin crestal ridge should address the probability that a buccal dehiscence will result. The aim of the present investigation was to perform guided bone regeneration (GBR) around implants with buccal dehiscences and evaluate the outcomes of using autogenous bone grafts harvested from three different intraoral sites. Forty-six Osseotite implants, 4 mm in diameter, were placed in thin crestal ridges, resulting in an uncovered implant surface from the buccal aspect. The lengths of the buccal dehiscences ranged from 3 to 7 mm as measured from the implant cervix to the most apical extent of the uncovered threads. A standard GBR technique was carried out to augment the bone defect around the buccal implant surface immediately after implant placement. The cases were divided into three groups according to receipt of an autogenous bone graft from the ramus, tuberosity, or mandibular symphysis. In all cases, e-PTFE membranes were used to cover the grafted areas. Grafted sites were exposed after 6 months, membranes were removed, and residual distance between the implant cervix and most uncovered thread was recorded. All grafting materials were able to produce a certain degree of bone regeneration. In terms of bone change level, the three groups were not equal. The mandibular symphysis group exhibited the highest mean bone growth level, followed by the ramus group. The tuberosity produced the poorest result. Mandibular and ramus autogenous bone grafts represent the best choice in materials for GBR procedures around implants, while tuberosity bone grafts can be used as an alternative. PMID- 15119887 TI - Use of the gingival unit transfer in soft tissue grafting: report of three cases. AB - Current procedures in periodontal plastic surgery probe the limits of the copious blood supply available to the periodontal tissues. None reaches for these limits more consistently and meets with less forgiveness than soft tissue grafting for the interdental papilla and root coverage in esthetically sensitive areas. Gingiva contains the only soft tissue naturally created to survive and function interproximally and facially over avascular root surfaces. Preliminary results suggest that the supracrestal gingiva, used as a free graft for the above purposes, may have an uncommon capacity for perfusion and survival. More investigation is necessary to confirm the validity of this concept. The gingival unit transfer is introduced through three case reports that describe its use for root coverage and replacement of intact but pathologically involved or esthetically compromised papillae. PMID- 15119888 TI - Sinus floor augmentation through a rotated palatal flap at the time of tooth extraction. AB - Rapid crestal bone resorption following maxillary tooth loss is further accentuated in the posterior regions because of pneumatization and enlargement of the maxillary sinuses. A treatment rationale that allows preservation and/or augmentation of vertical available bone at the time of posterior maxillary tooth extraction may offer numerous therapeutic benefits. The present study comprised 14 patients in whom 18 posterior maxillary teeth with no evident bone between the tooth apex and sinus floor, as estimated through preoperative radiographic analysis, were carefully extracted using a palatal approach. The empty alveolus was thoroughly debrided and incrementally filled with tricalcium phosphate. The graft material was gently pushed beyond the empty alveolus to elevate the sinus membrane using an osteotome. Primary soft tissue closure over the grafted sites was achieved by a rotated palatal flap. The distance between bone crest and sinus floor was radiographically estimated 6 to 7 months after the first procedure. Another procedure was then carried out to place the 10- to 14-mm implants, together with a bone-added osteotome sinus floor elevation. At uncovering, all implants were clinically stable, with no signs of infection. The presented surgical procedure performed at the time of extraction of posterior maxillary teeth in close proximity to the sinus floor allowed placement of implants of proper length and width, together with a bone-added osteotome sinus floor elevation during a second procedure. PMID- 15119889 TI - Clinical and radiographic evaluation of Emdogain as a regenerative material in the treatment of interproximal vertical defects in chronic and aggressive periodontitis patients. AB - This study evaluated the efficacy of Emdogain enamel matrix proteins as a regenerative material in interproximal vertical defects both clinically and radiographically. Patients aged 18 to 45 years and diagnosed with chronic or aggressive periodontitis were included. Sixteen intrabony defects in eight patients were surgically treated using a split-mouth design. Emdogain placement was done at experimental sites. Since both chronic and aggressive periodontitis patients were included, an attempt was made to interpret results between the two types of cases. Postsurgical measurements at 9 months revealed no significant difference in mean pocket depth reduction, clinical attachment level gain, amount of defect fill, or defect resolution between control and experimental groups. Mean pocket depth reduction and amount of defect fill were significant in both groups. The results were interpreted separately for chronic and aggressive periodontitis cases. This study demonstrated no added advantage of using Emdogain compared to surgical debridement alone. Further long-term and large-sample-size evaluation is required to prove Emdogain's consistent efficacy. PMID- 15119890 TI - Facing competition on veterinary medicines supply. PMID- 15119891 TI - A policy for bovine TB. PMID- 15119892 TI - Medicines, practice standards and VN training. PMID- 15119893 TI - Factors affecting the serological response of dogs and cats to rabies vaccination. AB - After being vaccinated against rabies some cats and dogs fail to show an antibody titre adequate to meet the requirements of the UK Pet Travel Scheme. To investigate this problem, the data derived from 16,073 serum samples submitted to the Veterinary Laboratories Agency for serological testing between 1999 and 2002, 1002 samples submitted to BioBest during March and April 2001, and 1264 samples associated with one make of vaccine submitted to BioBest between June 2001 and January 2003, were analysed. The probability of antibody titre failing to reach at least 0.5 iu/ml was analysed by logistic regression as a function of the choice of vaccine, the interval between vaccination and sampling, the sex and age of the animal, and its country of origin. In dogs, all these factors, except sex, had highly significant (P < 0.001) effects on the test failure rate, and in cats all the factors had a significant effect (P < 0.05). PMID- 15119894 TI - Radiographic findings in 16 dogs infected with Angiostrongylus vasorum. AB - Thoracic radiographs of 16 dogs infected naturally with Angiostrongylus vasorum showed signs of bronchial thickening, an interstitial pattern and a multifocal and/or peripheral alveolar pattern. In dogs treated with fenbendazole, follow-up radiographs showed that the alveolar pattern had resolved and a mild, hazy interstitial pattern had developed. In contrast with dogs with heartworm (Dirofilaria immitis), no pulmonary vascular lesions were identified. PMID- 15119895 TI - Diagnosis and management of B cell chronic lymphocytic leukaemia in a cat. AB - A four-year-old, female neutered domestic shorthair cat had a history of chronic intermittent vomiting and lymphocytosis. B cell chronic lymphocytic leukaemia was diagnosed by flow cytometry, which revealed abnormally large numbers of mature B lymphocytes in the peripheral blood. The cat was treated conservatively with antiemetic drugs and remained stable without chemotherapy for over a year. PMID- 15119896 TI - Ectoparasite and haemoparasite risks associated with imported exotic reptiles. PMID- 15119897 TI - Interferon treatment of circovirus infection in grey parrots (Psittacus e erithacus). PMID- 15119899 TI - Asymptomatic uterine rupture in a bitch. PMID- 15119898 TI - First record of Ornithonyssus bacoti from a domestic pet in the United Kingdom. PMID- 15119900 TI - Protein-losing enteropathy in a soft-coated wheaten terrier in the United Kingdom. PMID- 15119901 TI - Ischaemic necrosis of the base of the teat in dairy cows. PMID- 15119902 TI - Skin and feather disease in poultry of unknown aetiology. PMID- 15119903 TI - Mathematical approach to oxygen purchase. PMID- 15119904 TI - Clinical audit. PMID- 15119905 TI - Anticonvulsants as anxiolytics, part 2: Pregabalin and gabapentin as alpha(2)delta ligands at voltage-gated calcium channels. PMID- 15119906 TI - Sex and psychiatry in the next 5 years. PMID- 15119907 TI - Neuroleptic malignant syndrome and atypical antipsychotic drugs. AB - OBJECTIVE: The incidence of neuroleptic malignant syndrome (NMS) is not known, but the frequency of its occurrence with conventional antipsychotic agents has been reported to vary from 0.02% to 2.44%. DATA SOURCES: MEDLINE search conducted in January 2003 and review of references within the retrieved articles. DATA SYNTHESIS: Our MEDLINE research yielded 68 cases (21 females and 47 males) of NMS associated with atypical antipsychotic drugs (clozapine, N = 21; risperidone, N = 23; olanzapine, N = 19; and quetiapine, N = 5). The fact that 21 cases of NMS with clozapine were found indicates that low occurrence of extrapyramidal symptoms (EPS) and low EPS-inducing potential do not prevent the occurrence of NMS and D(2) dopamine receptor blocking potential does not have direct correlation with the occurrence of NMS. One of the cardinal features of NMS is an increasing manifestation of EPS, and the conventional antipsychotic drugs are known to produce EPS in 95% or more of NMS cases. With atypical antipsychotic drugs, the incidence of EPS during NMS is of a similar magnitude. CONCLUSIONS: For NMS associated with atypical antipsychotic drugs, the mortality rate was lower than that with conventional antipsychotic drugs. However, the mortality rate may simply be a reflection of physicians' awareness and ensuing early treatment. PMID- 15119908 TI - Managing atypical antipsychotic-associated weight gain: 12-month data on a multimodal weight control program. AB - BACKGROUND: The purpose of this study was to test prospectively the feasibility and efficacy of a multimodal weight control program for over-weight and obese severely mentally ill adults who had gained weight while taking atypical antipsychotic medications. METHOD: Thirty-one subjects with schizophrenia or schizoaffective disorder (DSM-IV), on treatment with atypical antipsychotics, participated in a 52-week, multimodal weight control program that incorporated nutrition, exercise, and behavioral interventions. The primary outcomes were measures of body mass index (BMI) and weight. A variety of secondary outcomes, including hemoglobin A(1c) level, systolic and diastolic blood pressure, and cholesterol level, were compared from baseline to endpoint. Weight and BMI changes in the intervention group were also compared with changes in 20 nonintervention patients ("usual care" group) who were contemporaneously treated in the same clinics. RESULTS: Twenty of the 31 subjects in the intervention group completed the program. Statistically significant pre-post improvements in weight (p <.02), BMI (p <.02), hemoglobin A(1c) levels (p <.001), diastolic (p <.001) and systolic (p <.05) blood pressure, exercise level (p <.003), nutrition knowledge (p <.0001), and stage of change (exercise [p <.0001] and weight [p <.008]) were seen in the intervention group. Patients attended a mean of 69% of the sessions during the year of the program. Weight and BMI also decreased significantly (p =.01) in the intervention group compared with the "usual care" group, who gained weight during the observation period. CONCLUSIONS: Individuals with schizophrenia and schizoaffective disorder were willing to attend, and benefited from, a weight control program that focused on nutrition, exercise, and motivation. The program resulted in clinically significant reductions in weight, BMI, and other risk factors for long-term poor health, including hemoglobin A(1c). In contrast, patients who did not receive the weight control intervention continued to gain weight. PMID- 15119909 TI - A multicenter, randomized, double-blind, placebo-controlled trial of extended release carbamazepine capsules as monotherapy for bipolar disorder patients with manic or mixed episodes. AB - BACKGROUND: Carbamazepine has been used to treat mania for over 2 decades. Most evaluations of carbamazepine have had important limitations, such as absence of a parallel placebo group, small sample size, or the confounding influence of concomitant treatment. All studies have used conventional, immediate-release carbamazepine formulations. We assessed the efficacy and safety of monotherapy with beaded, extended-release carbamazepine capsules (ERC-CBZ; SPD417) in bipolar disorder patients with manic or mixed episodes. METHOD: Following a single-blind placebo lead-in, DSM-IV-defined bipolar disorder patients with manic or mixed episodes were randomly assigned to receive ERC-CBZ (N = 101) or placebo (N = 103) for 3 weeks. Patients were hospitalized through the first 7 days of the double blind period. ERC-CBZ was initiated at 400 mg/day and increased, as necessary and tolerated, up to 1600 mg/day. Efficacy was assessed weekly with the Young Mania Rating Scale (YMRS), Clinical Global Impressions scale (CGI), and Hamilton Rating Scale for Depression (HAM-D). Data were gathered from December 1999 to June 2001. RESULTS: Ninety-six (47.1%) of 204 patients completed the study. The mean +/- SD final ERC-CBZ dose was 756.44 +/- 413.38 mg/day with a mean plasma drug level of 8.9 microg/mL. Starting at week 2, ERC-CBZ was associated with significantly greater improvements in YMRS (p =.032) using last-observation-carried-forward analyses. At end point, the responder rate (patients with at least a 50% decrease in YMRS score) also favored ERC-CBZ (41.5% vs. 22.4%; p =.0074). In a post hoc analysis of mixed patients, HAM-D score was significantly improved in patients remaining on ERC-CBZ treatment on day 21 (p =.01). Adverse events occurring more frequently in the ERC-CBZ group than in the placebo group included dizziness, nausea, and somnolence. CONCLUSION: We found ERC-CBZ to be effective in the first large, randomized, double-blind, placebo-controlled parallel trial of carbamazepine monotherapy in acute mania. This trial provides important additional evidence supporting the use of carbamazepine in acute mania. PMID- 15119910 TI - Speed of response and remission in major depressive disorder with acute electroconvulsive therapy (ECT): a Consortium for Research in ECT (CORE) report. AB - BACKGROUND: Remission of illness in patients with major depressive disorder (MDD) is achieved in less than half of patients initially treated with medication. Electroconvulsive therapy (ECT) is another treatment option. We report the speed of response and remission rates in a cohort of depressed patients who received a course of acute-phase ECT in the initial phase of an ongoing multicenter randomized trial of continuation ECT versus pharmacotherapy. METHOD: Patients with MDD according to DSM-IV criteria received bilateral ECT 3 times weekly. Prior to each treatment, a 24-item Hamilton Rating Scale for Depression (HAM-D 24) score was obtained by a clinical rater. Sustained response was defined as a > or = 50% reduction in baseline HAM-D-24 score for at least 2 and all subsequent measurement occasions. Remission was defined as HAM-D-24 scores of < or = 10 for at least the last 2 consecutive assessments. Data were collected from May 1997 through November 2000. RESULTS: Of the 253 patients who entered the study, 86% (N = 217) completed the acute course of ECT. Sustained response occurred in 79% of the sample, and remission occurred in 75% of the sample (N = 253); 34% (85/253) of patients achieved remission at or before ECT #6 (week 2), and 65% (164/253) achieved remission at or before ECT #10 (weeks 3-4). Over half (54%; 136/253) had an initial first response by ECT #3 (end of week 1). CONCLUSION: ECT was associated with rapid response and remission in a high percentage of patients. ECT warrants early consideration in treatment algorithms for patients with MDD. PMID- 15119911 TI - Hippocampal and amygdala changes in patients with major depressive disorder and healthy controls during a 1-year follow-up. AB - BACKGROUND: Although the hippocampus has been found to be smaller in patients with depression, prospective longitudinal in vivo studies are necessary to investigate whether depression can result in a further diminution of hippocampal volumes or whether a smaller hippocampal volume predisposes an individual to the development of depression. METHOD: Thirty patients with DSM-IV major depressive disorder as well as 30 healthy control subjects matched for age, gender, and handedness were examined at admission to the hospital and 1 year later using a documentation of the medical history and high-resolution magnetic resonance imaging (MRI) for the presence of depression and to determine changes in hippocampal as well as amygdala volumes. Patients were enrolled from March 2000 to August 2002. RESULTS: No significant hippocampal and amygdala volume changes were observed in patients or controls between baseline and 1-year follow-up investigations. However, the subgroup of patients who were nonremitted at the time of the follow-up investigation showed significantly reduced left and right hippocampal volumes at both baseline and the 1-year follow-up compared with remitted patients. Moreover, the right hippocampal volumes of nonremitted patients were significantly smaller compared with matched healthy controls. CONCLUSION: These results do not support the hypothesis that hippocampal volumes diminish during the 1-year follow-up period. However, smaller hippocampal volumes may be related to a poor clinical outcome after 1 year. PMID- 15119912 TI - The Texas Medication Algorithm Project antipsychotic algorithm for schizophrenia: 2003 update. AB - BACKGROUND: The Texas Medication Algorithm Project (TMAP) has been a public academic collaboration in which guidelines for medication treatment of schizophrenia, bipolar disorder, and major depressive disorder were used in selected public outpatient clinics in Texas. Subsequently, these algorithms were implemented throughout Texas and are being used in other states. Guidelines require updating when significant new evidence emerges; the antipsychotic algorithm for schizophrenia was last updated in 1999. This article reports the recommendations developed in 2002 and 2003 by a group of experts, clinicians, and administrators. METHOD: A conference in January 2002 began the update process. Before the conference, experts in the pharmacologic treatment of schizophrenia, clinicians, and administrators reviewed literature topics and prepared presentations. Topics included ziprasidone's inclusion in the algorithm, the number of antipsychotics tried before clozapine, and the role of first generation antipsychotics. Data were rated according to Agency for Healthcare Research and Quality criteria. After discussing the presentations, conference attendees arrived at consensus recommendations. Consideration of aripiprazole's inclusion was subsequently handled by electronic communications. RESULTS: The antipsychotic algorithm for schizophrenia was updated to include ziprasidone and aripiprazole among the first-line agents. Relative to the prior algorithm, the number of stages before clozapine was reduced. First generation antipsychotics were included but not as first-line choices. For patients refusing or not responding to clozapine and clozapine augmentation, preference was given to trying monotherapy with another antipsychotic before resorting to antipsychotic combinations. CONCLUSION: Consensus on algorithm revisions was achieved, but only further well-controlled research will answer many key questions about sequence and type of medication treatments of schizophrenia. PMID- 15119913 TI - Suicide attempts and ideation in patients with bipolar I disorder. AB - BACKGROUND: Suicidal thinking and behavior are common in individuals with bipolar disorder. METHOD: Suicidal ideation and suicide attempts were examined in 175 patients with bipolar I disorder (diagnosis confirmed with the Schedule for Affective Disorders and Schizophrenia or the Structured Clinical Interview for DSM-IV) participating in the Pittsburgh Study of Maintenance Therapies in Bipolar Disorder. Patients who attempted suicide before entering the study were compared with those who did not attempt suicide with respect to clinical and demographic characteristics. RESULTS: Twenty-nine percent of the patients had attempted suicide prior to entering the study. Suicide attempts happened at a relatively young age and in the first period of the illness. Greater severity of bipolar disorder, as indicated by a greater number of previous depressive episodes (p =.0009) and higher HAM-D-25 scores (p =.04), and higher body mass index (p =.03) were significantly correlated with a history of suicide attempts. None of the patients with a history of suicide attempt attempted suicide again. However, 5 patients without a history of suicide attempt did attempt suicide. Four of these patients did not display severe suicidal ideation at the assessment that preceded the suicide attempt. No subject completed suicide during the 11 years of the study. CONCLUSION: Greater severity of bipolar disorder and higher body mass index are significantly correlated with a history of suicide attempts. However, a treatment program in a maximally supportive clinical environment can reduce suicidal behavior in high-risk patients. In some cases, suicide risk is transient and may be preceded by a period of severe suicidal ideation that lasts only a few minutes or hours. In such cases, mental health professionals are unable to predict suicide attempts. PMID- 15119914 TI - Effect of reboxetine on major depressive disorder in breast cancer patients: an open-label study. AB - BACKGROUND: Depression is a common disorder in cancer patients, and it is associated with reduced quality of life, abnormal illness behavior, pain, and suicide risk. A few studies have investigated the effects of tricyclic antidepressants and serotonin reuptake inhibitors in cancer patients. No data are available regarding the use of reboxetine, a norepinephrine reuptake inhibitor that has been shown to be safe (e.g., absence of clinically significant drug-drug interactions and cytochrome P450 metabolism) and effective in the treatment of depressed patients, including those with medical illness (e.g., Parkinson's disease, human immunodeficiency virus infection). METHOD: The effects of reboxetine were investigated in 20 breast cancer patients with a DSM-IV diagnosis of major depressive disorder in an open, prospective 8-week trial. Severity of depression was assessed with the 17-item Hamilton Rating Scale for Depression (HAM-D). Psychiatric symptoms (Brief Symptom Inventory [BSI]), styles of coping with cancer (Mini-Mental Adjustment to Cancer [Mini-MAC]), quality of life (European Organization for Research and Treatment of Cancer Quality-of-Life Questionnaire C30 [EORTC-QLQ-C30]), and Clinical Global Impressions scale scores were also monitored. RESULTS: At 8 weeks, a significant (p <.01) reduction was observed in HAM-D scores, several BSI dimension scores, and Mini-MAC hopelessness and anxious preoccupation scores. A significant (p <.05) improvement from baseline to endpoint was found on the EORTC-QLQ-C30 subfactors emotional, cognitive, dyspnea, sleep, and global. Discontinuation was necessary in 1 subject because of hypomanic switch and in another because of side effects (tachycardia, tension). Seven patients experienced transient side effects (e.g., mild anxiety, insomnia, sweating). CONCLUSION: In this open trial, reboxetine appeared to be well tolerated and promising in reducing depressive symptoms and maladjusted coping styles and in improving scores on quality-of-life parameters. PMID- 15119915 TI - The effect of duloxetine on painful physical symptoms in depressed patients: do improvements in these symptoms result in higher remission rates? AB - BACKGROUND: Depression is a chronic disease consisting of emotional/psychological and physical symptoms. Emotional symptoms have been shown to respond to currently available antidepressants; however, physical symptoms may not be as responsive. It was hypothesized that resolution of both psychological and physical symptoms of depression would predict a higher percentage of patients achieving remission. METHOD: Efficacy data were pooled from 2 identical, but independent, 9-week randomized, double-blind clinical trials of duloxetine 60 mg q.d. (N = 251) and placebo (N = 261). All patients met diagnostic criteria for DSM-IV major depressive disorder, which was confirmed by the Mini-International Neuropsychiatric Interview. Efficacy measures included the 17-item Hamilton Rating Scale for Depression (HAM-D-17) total score, the HAM-D-17 Maier subscale, the Clinical Global Impressions-Severity of Illness (CGI-S) scale, the Patient Global Impression of Improvement (PGI-I) scale, the Somatic Symptom Inventory, the Quality of Life in Depression Scale, and Visual Analog Scales (VAS) for pain (overall pain, headaches, back pain, shoulder pain, interference with daily activities, and time in pain while awake). RESULTS: Duloxetine-treated patients demonstrated significantly greater improvement in overall pain (p =.016), back pain (p =.002), and shoulder pain (p =.021) at week 9 compared with patients receiving placebo. When treatment effects were pooled over all visits, patients receiving duloxetine, 60 mg q.d., exhibited significantly greater improvement than placebo-treated patients in 5 of the 6 assessed VAS pain measures. Approximately 50% of the improvement in overall pain was independent of improvement in HAM-D-17 total score. Assuming the same level of improvement in core emotional symptoms of depression (Maier subscale), improvement in overall pain severity was associated with higher estimated probabilities of remission (p <.001). The week 9 means for VAS overall pain severity were 13.0 for remitters (last observed value for HAM-D-17 was < or = 7) compared with 22.7 for nonremitters (p <.001), respectively, representing a greater than 3-fold improvement from baseline in remitters. The remission rate for pain responders (improvement in VAS overall pain from baseline to last observation > or = 50%) was twice that observed for pain nonresponders (36.2% vs. 17.8%, p <.001). Greater improvements in pain outcomes were associated with more favorable endpoint outcomes on the CGI-S and PGI-I scales. In addition, early favorable responses in VAS overall pain severity were associated with favorable endpoint outcomes. CONCLUSIONS: Treatment with duloxetine, 60 mg q.d., significantly reduced pain compared with placebo. Improvements in pain severity were attributable equally to the direct effect of duloxetine and to associated changes in depression severity. Improvement in painful physical symptoms was associated with higher remission rates even after accounting for improvement in core emotional symptoms. PMID- 15119916 TI - Health-related quality of life in patients with schizophrenia during treatment with long-acting, injectable risperidone. AB - BACKGROUND: We investigated the impact of treatment with long-acting, injectable risperidone versus placebo on health-related quality of life (HRQoL) in patients with schizophrenia. Results are discussed in the context of HRQoL in the general U.S. population. METHOD: Patients with DSM-IV schizophrenia entered a randomized, double-blind, placebo-controlled trial. After screening, previous antipsychotics were discontinued, and oral risperidone was titrated up to a dose of 4 mg/day over 1 week. Patients were then randomly assigned to receive placebo [N = 92] or long-acting risperidone (25 [N = 93], 50 [N = 97], or 75 mg [N = 87] every 2 weeks) for 12 weeks. HRQoL was measured using the Medical Outcomes Study Short Form 36-item questionnaire (SF-36). RESULTS: At week 12, patients receiving long acting risperidone had improved significantly (p <.05) in 5 domains of the SF-36 (bodily pain, general health, social functioning, role-emotional, and mental health) compared with patients receiving placebo. The effect was greatest for the 25-mg group, with significant improvement versus placebo in 6 domains (p <.05). At baseline, all SF-36 domain scores except bodily pain were significantly lower (p <.05) than normal values in all groups. With placebo, scores in all 8 domains remained below normal values after 12 weeks, while patients receiving long-acting risperidone showed improvement in HRQoL toward normal levels, with clinically meaningful improvements in all mental-health domains. In the 25-mg group, scores in 7 domains were not statistically different from normal values after 12 weeks. CONCLUSIONS: Long-acting, injectable risperidone improved HRQoL toward normal levels. After 12 weeks, HRQoL of patients receiving 25 mg was not significantly different from normal. PMID- 15119917 TI - Lack of effect of intravenous immunoglobulins on tics: a double-blind placebo controlled study. AB - BACKGROUND: Case studies and a placebo-controlled study previously suggested the effectiveness of immunomodulatory therapy in patients with tic or related disorders whose symptoms show a relationship with streptococcal infections. No data are available on the effectiveness of intravenous immunoglobulins (IVIG) on tic severity in unselected tic disorder patients. METHOD: Thirty patients with a DSM-IV tic disorder were randomly assigned to IVIG (1 g/kg on 2 consecutive days; mean age = 28.71 years; range, 14-53 years) or placebo (mean age = 30.73 years; range, 14-63 years). Symptoms were rated with the Yale Global Tic Severity Scale, the Yale-Brown Obsessive Compulsive Scale, and the Clinical Global Impressions scale of symptom change with regard to tic severity. These were used at baseline and on weeks 2, 4, 6, 10, and 14 posttreatment, after which blinding was broken. The study was conducted from March through August 2002. RESULTS: We observed no significant differences between both treatment groups regarding posttreatment changes in tic severity. Severity of obsessions and compulsions, which was in the subclinical range, decreased significantly in the IVIG group compared with the placebo group at week 6 (p =.02). Then, there was a 32.3% improvement in the IVIG group compared with baseline. Though this improvement was maintained over the following 8 weeks, no statistically significant differences between the IVIG and the placebo group with regard to improvements in obsessions and compulsions were detected at subsequent assessments. IVIG treatment was associated with significantly more side effects than placebo, most notably headache. CONCLUSION: Based on the present results, IVIG cannot be recommended in tic disorders. PMID- 15119918 TI - Somatic symptoms as predictors of time to onset of response to fluoxetine in major depressive disorder. AB - OBJECTIVE: In the present study we assessed the relationship between somatic symptoms and the time to onset of clinical response to fluoxetine in patients with major depressive disorder (MDD). METHOD: 87 outpatients (mean age = 41.4 +/- 10.2 years; 59.8% women) with DSM-III-R MDD who had sustained acute response to fluoxetine completed the Symptom Questionnaire (SQ) at baseline. Onset of response was defined as a 30% decrease in the total score for the 17-item Hamilton Rating Scale for Depression that led to a 50% decrease by week 8. With the use of 2 separate multiple regressions, controlling for the severity of depression at baseline, we then assessed the relationship between the number of somatic symptoms as assessed by the SQ subscale for somatic symptoms (SQ-SS) and both the time to onset of clinical response and the time to clinical response. The study was conducted between November 1992 and January 1999. RESULTS: A greater number of somatic symptoms at baseline predicted a greater amount of time to onset of clinical response to fluoxetine (p =.0233). The relationship between SQ-SS scores and time to response was not found to be statistically significant (p >.05). CONCLUSION: Somatic symptoms of depression were found to be associated with a delayed onset of antidepressant response to fluoxetine in MDD. PMID- 15119919 TI - Hyperlipidemia in persons using antipsychotic medication: a general population based birth cohort study. AB - BACKGROUND: Shortly after phenothiazines were introduced, they were found to elevate serum triglyceride and total cholesterol levels. During the past decade, an increasing body of literature has also documented this effect in atypical antipsychotics. Previous studies of antipsychotic-associated hyperlipidemias are based on clinical samples, mostly from case series. We studied the prevalence of hyperlipidemia in subjects who did and did not take antipsychotic medication in a prospective, general population-based birth cohort. METHOD: The study sample consisted of 5654 members of the unselected Northern Finland 1966 Birth Cohort who participated in the 1997-1998 clinical examination at 31 years of age. Blood samples were taken after an overnight fast, and serum total cholesterol, high density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL) cholesterol, and triglyceride levels were determined. Health habits and other possible correlates for hyperlipidemia were assessed using a questionnaire. The sample was analyzed in 4 categories according to use of antipsychotic medication: (1) atypical, (2) typical, (3) atypical and typical (for the 3 antipsychotic categories, total N = 45), and (4) no antipsychotic medication (N = 5609). Nonparametric tests and multiple logistic regression analysis were used to measure the effect of antipsychotics on serum lipids. RESULTS: High lipid levels were found in persons treated with both atypical and typical medication (mean total cholesterol = 233 mg/dL, mean triglycerides = 163 mg/dL). Mean total cholesterol and triglycerides were also high in subjects who used only typical medication (215 mg/dL and 148 mg/dL, respectively). The prevalence of hypercholesterolemia, high LDL cholesterol, and hypertriglyceridemia was high in persons using antipsychotic medication (31.1%, 20.0%, and 22.2%, respectively) compared with persons not using such medication (12.2%, 10.2%, and 7.0%, respectively). After we adjusted for risk factors for hyperlipidemia (sex, diet, waist circumference, physical exercise, smoking, and alcohol consumption), the results of logistic regression analysis showed that in persons treated with antipsychotic medication the risk of hypercholesterolemia was 2.8 (95% CI = 1.4 to 5.6); of hypertriglyceridemia, 2.3 (95% CI = 1.0 to 5.4); and of high LDL cholesterol, 1.6 (95% CI = 0.7 to 3.5). CONCLUSION: Lipid levels in subjects who used both atypical and typical medication and those who used only typical medication were high even in young age. As these persons are at special risk of hyperlipidemia, their lipid levels should be regularly monitored, and a cholesterol-lowering diet, as well as medication, should be considered. The results indicate an elevated risk of hyperlipidemia in persons using antipsychotic medication independent of the other risk factors assessed. PMID- 15119920 TI - Effects of atypical antipsychotics on the syndromal profile in treatment resistant schizophrenia. AB - BACKGROUND: There has been considerable support for the observation that atypical antipsychotics have a broader range of therapeutic effects than traditional antipsychotics. We are exploring whether this expanded clinical efficacy can also be seen in patients with treatment-resistant schizophrenia. METHOD: The subjects were 157 treatment-resistant inpatients diagnosed with DSM-IV schizophrenia or schizoaffective disorder. They were randomly assigned to treatment with clozapine, olanzapine, risperidone, or haloperidol in a 14-week double-blind trial and rated with a standard measure of clinical antipsychotic efficacy (Positive and Negative Syndrome Scale [PANSS]). Factor analysis at baseline and endpoint together with changes in 5 PANSS-derived factors were examined. Data were gathered from June 1996 to December 1999. RESULTS: The underlying PANSS factor structure, as indicated by the factor loadings, was essentially identical at baseline and endpoint. At baseline, the excitement factor was followed by the positive, negative, cognitive, and depression/anxiety factors, explaining 49.4% of the total variance. At endpoint, the positive factor was followed by the negative, excitement, cognitive, and depression/anxiety factors, explaining 55.5% of the total variance. The endpoint data indicated statistically significant (p <.05) improvements over time on the positive factor for all 3 atypicals, but not for haloperidol. The negative factor showed significant improvement for clozapine and olanzapine, with significant worsening for haloperidol. Clozapine, olanzapine, and risperidone were superior to haloperidol on the negative factor, while clozapine was also superior to risperidone. The cognitive factor showed significant improvement for all atypicals, as did the depression/anxiety factor. Only clozapine showed improvement on the excitement factor and was superior to both haloperidol and risperidone. CONCLUSIONS: Treatment with atypical antipsychotics did not substantially change the underlying PANSS 5-factor structure. However, antipsychotic treatment with all 3 atypical medications was associated with significant improvements on 3 of 5 syndromal domains (positive, cognitive, and depression/anxiety) of schizophrenia. Clozapine and olanzapine also showed improvement on the negative factor. Only clozapine was associated with improvement on the excitement domain. This finding confirms that atypicals are associated with improvement of an expanded spectrum of symptoms in treatment resistant patients. PMID- 15119922 TI - Augmentation of serotonin reuptake inhibitors in refractory obsessive-compulsive disorder using adjunctive olanzapine: a placebo-controlled trial. AB - BACKGROUND: The purpose of this study was to explore the efficacy of adding an atypical antipsychotic, olanzapine, to a serotonin reuptake inhibitor (SRI) in treatment-refractory obsessive-compulsive disorder (OCD). METHOD: Twenty-six patients aged between 18 and 65 (mean = 41.2, SD = 11.9) years meeting DSM-IV criteria for OCD, who had not responded to SRIs, were treated for 6 weeks in a double-blind, placebo-controlled augmentation study with either olanzapine (up to 20 mg/day) or placebo. Severity of illness was assessed biweekly by the Yale Brown Obsessive Compulsive Scale (Y-BOCS). Analysis of covariance with baseline Y BOCS score included as a covariate was used to compare improvement in Y-BOCS scores in the 2 groups. Response was defined as a 25% or greater improvement in Y BOCS score. Data were collected between April 2001 and May 2003. RESULTS: Outcome was assessed for all patients using the last observation carried forward. Subjects in the olanzapine group had a mean decrease of 4.2 (SD = 7.9) in Y-BOCS score compared with a mean increase in score of 0.54 (SD = 1.31) for subjects in the placebo group (F = 4.85, df = 2,23; p =.04). Six (46%) of 13 subjects in the olanzapine group showed a 25% or greater improvement in Y-BOCS score compared with none in the placebo group. The final mean dose of olanzapine was 11.2 (SD = 6.5) mg/day. Medication was well tolerated. Only 2 (15%) of 13 subjects who received olanzapine discontinued because of side effects: sedation (N = 1) or weight gain (N = 1). CONCLUSION: These results provide preliminary evidence that adding olanzapine to SRIs is potentially efficacious and well tolerated in the short-term treatment of patients with refractory OCD. Controlled studies with larger sample sizes are necessary to more definitively address this treatment strategy. PMID- 15119921 TI - Development of an atherogenic metabolic risk factor profile associated with the use of atypical antipsychotics. AB - BACKGROUND: It is important to assess cardiovascular risk factors to properly verify the potential consequences of atypical antipsychotic-related weight gain. The objective of the present study was to evaluate whether 2 atypical antipsychotics differ regarding their impact on the cardiovascular disease risk profile compared with a reference group. METHOD: We conducted a cross-sectional, multicenter study to assess anthropometric indices of obesity and to obtain a comprehensive fasting metabolic risk profile. Either risperidone or olanzapine had to be prescribed as the first and only antipsychotic for a minimum of 6 months. Patients were compared with a reference group of nondiabetic men. Data were collected from August 1999 to August 2001. RESULTS: Eighty-seven patients treated with olanzapine (N = 42) or risperidone (N = 45) were evaluated. Olanzapine-treated patients had significantly higher plasma triglyceride concentrations (2.01 +/-1.05 vs. 1.34 +/-0.65 mmol/L, p < or =.05), lower high density lipoprotein (HDL)-cholesterol levels (0.92 +/-0.17 vs. 1.04 +/- 0.21 mmol/L, p < or =.05), higher cholesterol/HDL-cholesterol ratios (5.62 +/-1.70 vs. 4.50 +/- 1.44, p < or =.05), higher apolipoprotein B levels (1.07 +/- 0.35 vs. 0.92 +/- 0.27 g/L, p < or =.05), smaller low-density lipoprotein peak particle diameters (252.6 +/-4.1 vs. 255.2 +/-4.3 A, p <.01), and higher fasting insulin concentrations (103.9 +/- 67.6 vs. 87.5 +/- 56.1 pmol/L, p < or =.05) than risperidone-treated patients. Moreover, 33% of olanzapine-treated patients were carriers of 3 atherogenic features of the metabolic syndrome as opposed to a prevalence of only 11% of risperidone-treated patients. CONCLUSION: These results suggest that olanzapine-treated patients are characterized by a more deteriorated metabolic risk factor profile compared with risperidone-treated patients. These observations raise concerns about the potential differential long-term deleterious effects of some antipsychotics, such as olanzapine, on cardiovascular health. PMID- 15119923 TI - International Consensus Group on Bipolar I Depression Treatment Guidelines. PMID- 15119924 TI - Lack of association between plasma apolipoprotein E and suicide attempts. PMID- 15119925 TI - Ribavirin may be an important factor in IFN-induced neuropsychiatric effects. PMID- 15119927 TI - Olanzapine may cause delirium in geriatric patients. PMID- 15119929 TI - A rose by any other name. PMID- 15119928 TI - A case of tardive dystonia successfully managed with quetiapine. PMID- 15119931 TI - Molecular mechanisms of cardiac protection by adaptation to chronic hypoxia. AB - Effective protection of the heart against ischemia/reperfusion injury is one of the most important goals of experimental and clinical research in cardiology. Besides ischemic preconditioning as a powerful temporal protective phenomenon, adaptation to chronic hypoxia also increases cardiac tolerance to all major deleterious consequences of acute oxygen deprivation such as myocardial infarction, contractile dysfunction and ventricular arrhythmias. Although many factors have been proposed to play a potential role, the detailed mechanism of this long-term protection remains poorly understood. This review summarizes current limited evidence for the involvement of ATP-sensitive potassium channels, reactive oxygen species, nitric oxide and various protein kinases in cardioprotective effects of chronic hypoxia. PMID- 15119932 TI - Genetic analysis of "metabolic syndrome" in the spontaneously hypertensive rat. AB - In the current review, we summarize results of genetic analyses of "metabolic syndrome" in the spontaneously hypertensive rat (SHR). These results include (1) linkage analyses in the HXB/BXH recombinant inbred (RI) strains derived from SHR and Brown Norway (BN-Lx) strains which revealed quantitative trait loci (QTL) for hemodynamic and metabolic traits on several chromosomes, (2) genetic isolation of these putative QTL within differential chromosome segments of SHR.BN congenic strains, (3) detailed mapping of these QTL within limited chromosome segments of SHR.BN congenic sublines, (4) sequencing of selected positional candidate genes which revealed important mutations in the Cd36 and Srebp1 SHR genes, (5) functional tests of these candidate genes in SHR transgenic lines, and (6) integrated gene expression profiling and linkage mapping in RI strains which will be used to identify co-regulated genes and to determine co-segregation of transcriptional profiles with physiological and pathophysiological phenotypes. PMID- 15119933 TI - Altered balance of vasoactive systems in experimental hypertension: the role of relative NO deficiency. AB - This review summarizes our findings concerning the altered balance of vasoactive systems (namely sympathetic nervous system and nitric oxide) in various forms of experimental hypertension--genetic hypertension (SHR, HTG rats), salt hypertension (Dahl rats) and NO-deficient hypertension (L-NAME-treated rats). An attempt is made to define relative NO deficiency (compared to the existing level of sympathetic vasoconstriction), to describe its possible causes and to evaluate particular indicators of its extent. A special attention is paid to reactive oxygen species, their interaction with NO metabolism, cell Ca2+ handling and blood pressure regulation. Our current effort is focused on the investigation of abnormal regulation of cytosolic Ca2+ levels in smooth muscle and endothelium of hypertensive animals. Such a research should clarify the mechanisms by which genetic and/or environmental factors could chronically modify blood pressure level. PMID- 15119934 TI - Cell adhesion on artificial materials for tissue engineering. AB - Advanced interdisciplinary scientific field of tissue engineering has been developed to meet increasing demand for safe, functional and easy available substitutes of irreversibly damaged tissues and organs. First biomaterials were constructed as "two-dimensional" (allowing cell adhesion only on their surface), and durable (non-biodegradable). In contrast, biomaterials of new generation are characterized by so-called three dimensional porous or scaffold-like architecture promoting attachment, growth and differentiation of cells inside the material, accompanied by its gradual removal and replacement with regenerated fully functional tissue. In order to control these processes, these materials are endowed with a defined spectrum of bioactive molecules, such as ligands for adhesion receptors on cells, functional parts of natural growth factors, hormones and enzymes or synthetic regulators of cell behavior, incorporated in defined concentrations and spatial distribution against a bioinert background resistant to uncontrolled protein adsorption and cell adhesion. PMID- 15119935 TI - Confocal stereology and image analysis: methods for estimating geometrical characteristics of cells and tissues from three-dimensional confocal images. AB - A short review of confocal stereology and three-dimensional image analysis is presented, pointing out the achievements accomplished in this field by the Department of Biomathematics (Institute of Physiology, Prague). One of the methods of confocal stereology, the fakir method for surface area estimation, developed by this laboratory, is described. Methods for automatic measurement of geometrical characteristics of microscopical structures, based on 3-D image processing or surface triangulation, are discussed and compared with interactive stereological methods. Three-dimensional reconstruction programs and software implementation of stereological and digital methods as well as their practical applications are presented. The future trends are discussed. PMID- 15119936 TI - Influence of thyroid status on the differentiation of slow and fast muscle phenotypes. AB - Muscle phenotype is determined by combined effects of intrinsic genetic and extrinsic factors like innervation, hormonal levels and mechanical factors or muscle activity. We have been studying the effect of altered thyroid hormone levels on the expression of myosin heavy chain (MyHC) isoforms in control and regenerating soleus and extensor digitorum longus muscles of euthyroid, hypothyroid or hyperthyroid female inbred Lewis rats. The fiber type composition has been determined according to the mATPase activity and immunocytochemical staining of MyHC isoforms, the content of MyHC isoforms has been determined by SDS-PAGE, the mRNA levels have been measured by RT-PCR and the ultrastructural transformation has been analyzed by electron-microscopy. Our results indicate that although the innervation plays a decisive role in the determination of muscle phenotype, levels of thyroid hormones contribute to the extent of muscle phenotype transformation. PMID- 15119937 TI - Corticosteroid regulation of colonic ion transport during postnatal development: methods for corticosteroid analysis. AB - Many mammalian species including human are immature at birth and undergo major developmental changes during suckling and weaning period. This problem is also conspicuous for the gastrointestinal tract that undergoes abrupt transitions coinciding with birth and weaning. This review deals with the maturation of ion transport functions in colon, the intestinal segment that plays an important role in sodium and potassium absorption and secretion. The purpose of the present review is to summarize the mechanism of sodium and potassium transport pathways and show how these transport processes change postnatally and how hormones, particularly corticosteroids, modify the pattern of development. Finally we describe some of the ways, how to analyze corticosteroid metabolism in target tissue. PMID- 15119938 TI - Metabolism of bromide and its interference with the metabolism of iodine. AB - The present knowledge about the metabolism of bromide with respect to its goitrogenic effects, including some conclusions drawn from our recent research on this subject, is reviewed. Firstly, the biological behavior of bromide ion is compared with that of chloride and iodide. Secondly, the details about distribution and kinetics of bromide ions in the body and in 15 different organs and tissues of the rat are given. Significant correlation between the values of the steady-state concentration of bromide in the respective tissue and of the corresponding biological half-life was found in most tissues examined. A remarkably high concentration of radiobromide was found in the skin, which represents, due to its large mass, the most abundant depot of bromide in the body of the rat. Thirdly, the effects of excessive bromide on the rat thyroid are summarized, along with the interference of exogenous bromide with the whole-body metabolism of iodine. It is suggested that high levels of bromide in the organism of experimental animals can influence their iodine metabolism in two parallel ways: by a decrease in iodide accumulation in the thyroid and skin (and in the mammary glands in lactating dams), and by a rise in iodide excretion by kidneys. By accelerating the renal excretion of iodide, excessive bromide can also influence the pool of exchangeable iodide in the thyroid. Finally, our recent results concerning the influence of high bromide intake in the lactating rat dam on iodine and bromide transfer to the suckling, and the impact of seriously decreased iodine content and increased bromide concentration in mother's milk on the young are discussed. We must state, however, that the virtue of the toxic effects of excessive bromide on the thyroid gland and its interference with the biosynthesis of thyroid hormones, as well as the exact mechanism of bromide interference with postnatal developmental processes remains to be elucidated. PMID- 15119939 TI - Yeast as a model organism to study transport and homeostasis of alkali metal cations. AB - To maintain an optimum cytoplasmic K(+)/Na+ ratio, cells employ three distinct strategies: 1) strict discrimination among alkali metal cations at the level of influx, 2) efficient efflux of toxic cations from cells, and 3) selective sequestration of cations in organelles. Cation efflux and influx are mediated in cells by systems with different substrate specificities and diverse mechanisms, e.g. ATPases, symporters, antiporters, and channels. Simple eukaryotic yeast Saccharomyces cerevisiae cells proved to be an excellent model for studying the transport properties and physiological function of alkali-metal-cation transporters, and the existence of mutant strains lacking their own transport systems provided an efficient tool for a molecular study of alkali-metal-cation transporters from higher eukaryotes upon their expression in yeast cells. PMID- 15119940 TI - Down-regulation of model yeast proteins by ubiquitin-dependent proteolysis. AB - Ubiquitination is a versatile tool used by all eukaryotic organisms for controlling the stability, function, and intracellular localization of a multitude of proteins. I will attempt to bring together our recent data on the down-regulation of two yeast model proteins, the galactose transporter Gal2 and fructose-1,6-bisphosphatase, by ubiquitin-dependent proteolysis triggered by the addition of easily fermentable carbon sources. PMID- 15119941 TI - Activation and modulation of ligand-gated ion channels. AB - Ligand-gated ionic channels are integral membrane proteins that enable rapid and selective ion fluxes across biological membranes. In excitable cells, their role is crucial for generation and propagation of electrical signals. This survey describes recent results from studies performed in the Department of Cellular Neurophysiology, Institute of Physiology ASCR, aimed at exploring the conformational dynamics of the acetylcholine, glutamate and vanilloid receptors during their activation, inactivation and desensitization. Distinct families of ion channels were selected to illustrate a rich complexity of the functional states and conformational transitions these proteins undergo. Particular attention is focused on structure-function studies and allosteric modulation of their activity. Comprehension of the fundamental principles of mechanisms involved in the operation of ligand-gated ion channels at the cellular and molecular level is an essential prerequisite for gaining an insight into the pathogenesis of many psychiatric and neurological disorders and for efficient development of novel specifically targeted drugs. PMID- 15119942 TI - Excitatory aminoacids and epileptic seizures in immature brain. AB - Data on convulsant and anticonvulsant action of drugs influencing excitatory amino acid receptors in developing rats are reviewed. Agonists of NMDA type of receptors NMDA and homocysteic acid, elicited an age-related seizure pattern- flexion, emprosthotonic seizures--in the first three postnatal weeks of rats. Generalized clonic-tonic seizures appeared only after a longer latency. Kainic acid administration resulted in epileptic automatisms and later in minimal, clonic seizures followed by generalized tonic-clonic seizures. A decrease of sensitivity to convulsant action with age is a general rule for all agonists tested. Different anticonvulsant action of NMDA and nonNMDA antagonists was demonstrated in a model of generalized tonic-clonic seizures induced by pentetrazol, whereas their action against epileptic afterdischarges elicited by electrical stimulation of cerebral cortex was similar. Again, higher efficacy in younger animals was a rule. As far as metabotropic glutamate receptors are concerned, agonists of groups II and III were shown to protect against convulsant action of homocysteic acid in immature rats and an antagonist of group I receptors MPEP suppressed the tonic phase of generalized tonic-clonic seizures induced by pentetrazol more efficiently in younger than in more mature rat pups. Unfortunately, a higher sensitivity to the action of antagonists of ionotropic glutamate receptors was demonstrated also for unwanted side effects (motor functions were compromized). In contrast, glutamate metabotropic receptor antagonist MPEP did not exhibit any serious side effects in rat pups. PMID- 15119943 TI - The role of dorsal columns pathway in visceral pain. AB - Traditionally, the dorsal column-medial lemniscus system has been viewed as a pathway not involved in pain perception. However, recent clinical and experimental studies have provided compelling evidence that implicates an important role of the dorsal column pathway in relaying visceral nociceptive information. Several clinical studies have shown that a small lesion that interrupts fibers of the dorsal columns (DC) that ascend close to the midline of the spinal cord significantly relieves pain and decreases analgesic requirements in patients suffering from cancer originating in visceral organs. Behavioral, electrophysiological and immunohistochemical methods used under experimental situations in animals showed that DC lesion lead to decreased activation of thalamic and gracile neurons by visceral stimuli, suppressed inhibition of exploratory activity induced by visceral noxious stimulation and prevented potentiation of visceromotor reflex evoked by colorectal distention under inflammatory conditions. Whereas the surgical lesion of the DC tract has proven to be clinically successful, a pharmacological approach would be a better strategy to block this pathway and thus to improve visceral pain conditions under less dramatic circumstances than cancer pain. Our finding that PSDC neurons start to express receptors for substance P after colon inflammation suggests new targets for the development of pharmacological strategies for the control of visceral pain. PMID- 15119944 TI - Regulation of signal transduction at M2 muscarinic receptor. AB - Muscarinic acetylcholine receptors mediate transmission of an extracellular signal represented by released acetylcholine to neuronal or effector cells. There are five subtypes of closely homologous muscarinic receptors which are coupled by means of heterotrimeric G-proteins to a variety of signaling pathways resulting in a multitude of target cell effects. Endogenous agonist acetylcholine does not discriminate among individual subtypes and due to the close homology of the orthosteric binding site the same holds true for most of exogenous agonists. In addition to the classical binding site muscarinic receptors have one or more allosteric binding sites at extracellular domains. Binding of allosteric modulators induces conformational changes in the receptor that result in subtype specific changes in orthosteric binding site affinity for both muscarinic agonists and antagonists. This overview summarizes our recent experimental effort in investigating certain aspects of M2 muscarinic receptor functioning concerning i) the molecular determinants that contribute to the binding of allosteric modulators, ii) G-protein coupling specificity and subsequent cellular responses and iii) possible functional assays that exploit the unique properties of allosteric modulators for characterization of muscarinic receptor subtypes in intact tissue. A detailed knowledge of allosteric properties of muscarinic receptors is required to permit drug design that will modulate signal transmission strength of specific muscarinic receptor subtypes. Furthermore, allosteric modulation of signal transmission strength is determined by cooperativity rather than concentration of allosteric modulator and thus reduces the danger of overdose. PMID- 15119945 TI - Biochemistry of transmembrane signaling mediated by trimeric G proteins. AB - Many extracellular signals are at the cell surface received by specific receptors, which upon activation transduce information to the appropriate cellular effector molecules via trimeric G proteins. The G protein-mediated cascades ultimately lead to the highly refined regulation of systems such as sensory perception, cell growth, and hormonal regulation. Transmembrane signaling may be seriously deranged in various pathophysiological conditions. Over the last two decades the major experimental effort of our group has been devoted to better understanding the molecular mechanisms underlying transmembrane signaling regulated by G proteins and to the closely related process of desensitization of hormone response. This review provides general information about the basic principles of G protein-regulated transmembrane signaling as well as about our contribution to the current progress in the field. PMID- 15119946 TI - Melatonin action in neonatal gonadotrophs. AB - Neonatal pituitary cells express MT1 and MT2 subtype of melatonin receptors that are coupled to pertussis toxin-sensitive G proteins. Their activation by melatonin leads to a decrease in cAMP production and activity of protein kinase A, and attenuation of gonadotropin-releasing hormone (GnRH)-induced gonadotropin secretion. Single cell calcium and electrophysiological recordings have revealed that a reduction in gonadotropin release results from melatonin-induced inhibition of GnRH-stimulated calcium signaling. Melatonin inhibits both calcium influx through voltage-dependent calcium channels and calcium mobilization from intracellular stores. Inhibition of calcium influx, probably in a cAMP/protein kinase C-dependent manner, and the accompanying calcium-induced calcium release from ryanodine-sensitive intracellular pools by melatonin results in a delay of GnRH-induced calcium signaling. Melatonin-induced attenuation of GnRH-induced and inositol (1,4,5)-trisphosphate-mediated calcium release from intracellular pools attenuates the amplitude of calcium signal. The potent inhibition of GnRH-induced calcium signaling and gonadotropin secretion by melatonin provides an effective mechanism to protect premature initiation of pubertal changes that are dependent on plasma gonadotropin levels. During the development, such tonic inhibitory effects of melatonin on GnRH action gradually decline due to a decrease in expression of functional melatonin receptors. In adult animals, melatonin does not have obvious direct effects on pituitary functions, whereas the connections between melatonin release and hypothalamic functions, including GnRH release, are preserved, and are critically important in synchronizing the external photoperiods and reproductive functions through still not well characterized mechanisms. PMID- 15119947 TI - Seasonal molecular timekeeping within the rat circadian clock. AB - In temperate zones duration of daylight, i.e. photoperiod, changes with the seasons. The changing photoperiod affects animal as well as human physiology. All mammals exhibit circadian rhythms and a circadian clock controlling the rhythms is located in the suprachiasmatic nucleus (SCN) of the hypothalamus. The SCN consists of two parts differing morphologically and functionally, namely of the ventrolateral (VL) and the dorsomedial (DM). Many aspects of SCN-driven rhythmicity are affected by the photoperiod. The aim of the present overview is to summarize data about the effect of the photoperiod on the molecular timekeeping mechanism in the rat SCN, especially the effect on core clock genes, clock-controlled genes and clock-related genes expression. The summarized data indicate that the photoperiod affects i) clock-driven rhythm in photoinduction of c-fos gene and its protein product within the VL SCN, ii) clock-driven spontaneous rhythms in clock-controlled, i.e. arginine-vasopressin, and in clock related, i.e. c-fos, gene expression within the DM SCN, and iii) the core clockwork mechanism within the rat SCN. Hence, the whole central timekeeping mechanism within the rat circadian clock measures not only the daytime but also the time of the year, i.e. the actual season. PMID- 15119948 TI - From spreading depression to spatial cognition. AB - The Laboratory of Neurophysiology of Memory started its existence in 1954 by systematic research into spreading depression of EEG activity of laboratory rodents and by the use of this remarkable phenomenon as a functional ablation method in behavioral research. Its main contributions were in the study of memory formation and consolidation, interhemispheric transfer, motor learning, conditioned taste aversion and spatial orientation and navigation. In the last five years it concentrated on navigation of rats in multiple reference frames, on electrophysiological evidence for the role of hippocampal place cells support of behavior in such dissociated frames, on the analysis of idiothetic and allothetic forms of navigation and on the mathematical methods allowing assessment of the contribution of goal directed locomotion to place cell activity. The methods used in spatial memory research in rats were used for examination of human subjects in a laboratory equipped with a tracking system for humans in the hospital Homolka. Animal models of Alzheimer disease were studied in transgenic mice with the human gene for the beta amyloid precursor protein. PMID- 15119949 TI - Protein modeling combined with spectroscopic techniques: an attractive quick alternative to obtain structural information. AB - Beside of the protein crystallography or NMR, another attractive option in protein structure analysis has recently appeared: computer modeling of the protein structure based on homology and similarity with proteins of already known structures. We have used the combination of computer modeling with spectroscopic techniques, such as steady-state or time-resolved fluorescence spectroscopy, and with molecular biology techniques. This method could provide useful structural information in the cases where crystal or NMR structure is not available. Molecular modeling of the ATP site within the H4-H5-loop revealed eight amino acids residues, namely besides the previously reported amino acids Asp443, Lys480, Lys501, Gly502 and Arg544, also Glu446, Phe475 and Gln482, which form the complete ATP recognition site. Moreover, we have proved that a hydrogen bond between Arg423 and Glu472 supports the connection of two opposite halves of the ATP-binding pocket. Similarly, the conserved residue Pro489 is important for the proper interaction of the third and fourth beta-strands, which both contain residues that take part in the ATP-binding. Alternatively, molecular dynamics simulation combined with dynamic fluorescence spectroscopy revealed that 14-3-3 zeta C-terminal stretch is directly involved in the interaction of 14-3-3 protein with the ligand. Phosphorylation at Thr232 induces a conformational change of the C-terminus, which is presumably responsible for observed inhibition of binding abilities. Phosphorylation at Thr232 induces more extended conformation of 14-3 3zeta C-terminal stretch and changes its interaction with the rest of the 14-3-3 molecule. This could explain negative regulatory effect of phosphorylation at Thr232 on 14-3-3 binding properties. PMID- 15119950 TI - Mitochondrial uncoupling proteins--facts and fantasies. AB - Instead of a comprehensive review, we describe the basic undisputed facts and a modest contribution of our group to the fascinating area of the research on mitochondrial uncoupling proteins. After defining the terms uncoupling, leak, protein-mediated uncoupling, we discuss the assumption that due to their low abundance the novel mitochondrial uncoupling proteins (UCP2 to UCP5) can provide only a mild uncoupling, i.e. can decrease the proton motive force by several mV only. Contrary to this, the highly thermogenic role of UCP1 in brown adipose tissue is not given only by its high content (approximately 5 % of mitochondrial proteins) but also by the low ATP synthase content and high capacity respiratory chain. Fatty acid cycling mechanism as a plausible explanation for the protonophoretic function of all UCPs and some other mitochondrial carriers is described together with the experiments supporting it. The phylogenesis of all UCPs, estimated UCP2 content in several tissues, and details of UCP2 activation are described on the basis of our experiments. Functional activation of UCP2 is proposed to decrease reactive oxygen species (ROS) production. Moreover, reaction products of lipoperoxidation such as cleaved hydroperoxy-fatty acids and hydroxy fatty acid can activate UCP2 and promote feedback down-regulation of mitochondrial ROS production. PMID- 15119951 TI - Genetic defects of cytochrome c oxidase assembly. AB - Cytochrome c oxidase (COX), the terminal enzyme of the mitochondrial respiratory chain, is one of the key functional and regulatory sites of the mammalian energy metabolism. Owing to the importance of the enzyme, pathogenetic mutations affecting COX frequently result in severe, often fatal metabolic disorders. No satisfactory therapy is currently available so that the treatment remains largely symptomatic and does not improve the course of the disease. While only few genetic defects of COX are caused by mutations in mitochondrial genome, during the last five years a large number of pathogenetic mutations in nuclear genes have been discovered. All these mutations are located in genes encoding COX specific assembly proteins including SURF1, SCO1, SCO2, COX10, and COX15. Despite the identification of increasing number of mutations, their precise etiopathogenetic mechanisms, which are necessary for the development of future therapeutic protocols, still remain to be elucidated. This review summarizes recent developments, including our efforts in elucidation of the molecular basis of human mitochondrial diseases due to specific defects of COX with special focus on SURF1 assembly protein. PMID- 15119952 TI - Energy metabolism of adipose tissue--physiological aspects and target in obesity treatment. AB - Body fat content is controlled, at least in part, by energy charge of adipocytes. In vitro studies indicated that lipogenesis as well as lipolysis depend on cellular ATP levels. Respiratory uncoupling may, through the depression of ATP synthesis, control lipid metabolism of adipose cells. Expression of some uncoupling proteins (UCP2 and UCP5) as well as other protonophoric transporters can be detected in the adipose tissue. Expression of other UCPs (UCP1 and UCP3) can be induced by pharmacological treatments that reduce adiposity. A negative correlation between the accumulation of fat and the expression of UCP2 in adipocytes was also found. Ectopic expression of UCP1 in the white fat of aP2 Ucp1 transgenic mice mitigated obesity induced by genetic or dietary factors. In these mice, changes in lipid metabolism of adipocytes were associated with the depression of intracellular energy charge. Recent data show that AMP-activated protein kinase may be involved in the complex changes elicited by respiratory uncoupling in adipocytes. Changes in energy metabolism of adipose tissue may mediate effects of treatments directed against adiposity, dyslipidemia, and insulin resistance. PMID- 15119953 TI - Simultaneous measurement of sensor-protein dynamics and motility of a single cell by on-chip microcultivation system. AB - Measurement of the correlation between sensor-protein expression, motility and environmental change is important for understanding the adaptation process of cells during their change of generation. We have developed a novel assay exploiting the on-chip cultivation system, which enabled us to observe the change of the localization of expressed sensor-protein and the motility for generations. Localization of the aspartate sensitive sensor protein at two poles in Escherichia coli decreased quickly after the aspartate was added into the cultivation medium. However, it took more than three generations for recovering the localization after the removal of aspartate from the medium. Moreover, the tumbling frequency was strongly related to the localization of the sensor protein in a cell. The results indicate that the change of the spatial localization of sensor protein, which was inherited for more than three generations, may contribute to cells, motility as the inheritable information. PMID- 15119954 TI - Applications of nanoparticles in biology and medicine. AB - Nanomaterials are at the leading edge of the rapidly developing field of nanotechnology. Their unique size-dependent properties make these materials superior and indispensable in many areas of human activity. This brief review tries to summarise the most recent developments in the field of applied nanomaterials, in particular their application in biology and medicine, and discusses their commercialisation prospects. PMID- 15119955 TI - The aryl hydrocarbon receptor-mediated disruption of vitellogenin synthesis in the fish liver: Cross-talk between AHR- and ERalpha-signalling pathways. AB - BACKGROUND: In the fish liver, the synthesis of egg yolk protein precursor vitellogenin (VTG) is under control of the estrogen receptor alpha (ERalpha). Environmental contaminants such as 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD) are suspected to have antiestrogenic effects. The aryl hydrocarbon receptor (AHR) is the initial cellular target for TCDD and related compounds. The AHR is a ligand-activated transcription factor that stimulates the expression of the genes encoding xenobiotic metabolizing enzymes, such as cytochrome P450 1A (CYP1A). In this study, the effects of activation of AHR on the hepatic expression of VTG and ERalpha genes, in primary cultured salmon hepatocytes, have been investigated. RESULTS: The expression of the genes encoding VTG and ERalpha were strongly induced by 17beta-estradiol (E2). However, the expression of VTG was disrupted by exposure of the cells to TCDD while CYP1A expression was enhanced. The effect of TCDD on VTG and CYP1A expression was annulled by the AHR-inhibitor alpha naphthoflavone. Furthermore, exposure of the cells to TCDD abolished E2-induced accumulation of ERalpha mRNA. The AHR-mediated inhibitory effects on the expression of the VTG and ERalpha genes may occur at transcriptional and/or post transcriptional levels. Nuclear run-off experiments revealed that simultaneous exposure of the cells to E2 and TCDD strongly inhibited the initiation of transcription of the VTG and ERalpha genes. In addition, inhibition of RNA synthesis by actinomycin D treatment showed that post-transcriptional levels of VTG and ERalpha mRNAs were not significantly altered upon treatment of the cells with TCDD. These results suggested that activation of AHR may inhibit the transactivation capacity of the ERalpha. Further, electrophoretic mobility shift assays using nuclear extracts prepared from cells treated for one or two hours with E2, alone or in mixture with TCDD, showed a strong reduction in the DNA binding activities upon TCDD treatment. These results also suggested that activation of the AHR signalling pathway caused a marked decrease in the number of the nuclear ERalpha or that activated AHR blocked the ability of ERalpha to bind to its target DNA sequence. Finally, our results from Northern hybridizations indicated that E2 treatment of the cells did not cause any significant effect on the TCDD-induced levels of CYP1A mRNA. CONCLUSION: In fish hepatocytes E2 induces ERalpha and VTG gene expression. The presence of dioxin (TCDD) abolishes this induction, probably through the action of AHR in complex with AHR nuclear translocator, and possibly by direct interference with the auto regulatory transcriptional loop of ERalpha. Furthermore, E2 does not interfere with TCDD induced CYP1A gene expression, suggesting that cross-talk between the ERalpha- and AHR-signalling pathways is unidirectional. PMID- 15119957 TI - Activation of the control reporter plasmids pRL-TK and pRL-SV40 by multiple GATA transcription factors can lead to aberrant normalization of transfection efficiency. AB - BACKGROUND: Members of the GATA transcription factor family have been used in many transfection studies to investigate their roles in the regulation of gene expression. To correct for variations in transfection efficiency, the Renilla luciferase encoding plasmids pRL-TK and pRL-SV40 are commonly used as normalization controls. RESULTS: We report here that plasmids expressing GATA-4 or GATA-6 transcription factor increased Renilla luciferase gene expression by 2 to 8 fold when co-transfected with pRL-TK or pRL-SV40. This alteration of the control reporter gene activity was shown to cause erroneous normalization of transfection efficiency and thus misinterpretation of results in a transactivation assay. To circumvent the problem, we generated two mutant control plasmids from which putative GATA response elements were deleted. These deletions rendered pRL-SV40 unresponsive to GATA transcription factor stimulation and reduced the response of pRL-TK. A database search also indicates that consensus GATA binding sequences are present in other commercially available Renilla luciferase encoding plasmids; therefore, the latter can potentially be transactivated by GATA transcription factors. CONCLUSION: Taken together, these findings highlight the importance of the selection of an appropriate control reporter plasmid for the normalization of transfection efficiency. PMID- 15119956 TI - S-allylmercaptocysteine scavenges hydroxyl radical and singlet oxygen in vitro and attenuates gentamicin-induced oxidative and nitrosative stress and renal damage in vivo. AB - BACKGROUND: Oxidative and nitrosative stress have been involved in gentamicin induced nephrotoxicity. The purpose of this work was to study the effect of S allylmercaptocysteine, a garlic derived compound, on gentamicin-induced oxidative and nitrosative stress and nephrotoxicity. In addition, the in vitro reactive oxygen species scavenging properties of S-allylmercaptocysteine were studied. RESULTS: S-allylmercaptocysteine was able to scavenge hydroxyl radicals and singlet oxygen in vitro. In rats treated with gentamicin (70 mg/Kg body weight, subcutaneously, every 12 h, for 4 days), renal oxidative stress was made evident by the increase in protein carbonyl content and 4-hydroxy-2-nonenal, and the nitrosative stress was made evident by the increase in 3-nitrotyrosine. In addition, gentamicin-induced nephrotoxicity was evident by the: (1) decrease in creatinine clearance and in activity of circulating glutathione peroxidase, and (2) increase in urinary excretion of N-acetyl-beta-D-glucosaminidase, and (3) necrosis of proximal tubular cells. Gentamicin-induced oxidative and nitrosative stress and nephrotoxicity were attenuated by S-allylmercaptocysteine treatment (100 mg/Kg body weight, intragastrically, 24 h before the first dose of gentamicin and 50 mg/Kg body weight, intragastrically, every 12 h, for 4 days along gentamicin-treatment). CONCLUSION: In conclusion, S-allylmercaptocysteine is able to scavenge hydroxyl radicals and singlet oxygen in vitro and to ameliorate the gentamicin-induced nephrotoxicity and oxidative and nitrosative stress in vivo. PMID- 15119958 TI - Study of the distribution of Malassezia species in patients with pityriasis versicolor and healthy individuals in Tehran, Iran. AB - BACKGROUND: Pityriasis versicolor is a superficial infection of the stratum corneum which caused by a group of yeasts formerly named pityrosporium. The taxonomy of these lipophilic yeasts has recently been modified and includes seven species referred as Malassezia. The aim of this study is to compare the distribution of Malassezia species isolated from pityriasis versicolor lesions and those isolated from healthy skins. METHODS: Differentiation of all malassezia species performed using morphological features and physiological test including catalase reaction, Tween assimilation test and splitting of esculin. RESULTS: In pityriasis versicolor lesions, the most frequently isolated species was M. globosa (53.3%), followed by M. furfur (25.3%), M. sympodialis(9.3%), M. obtusa (8.1%) and M. slooffiae (4.0%). The most frequently isolated species in the skin of healthy individuals were M. globosa, M. sympodialis, M. furfur, M. sloofiae and M. restricta which respectively made up 41.7%, 25.0%, 23.3%, 6.7% and 3.3% of the isolated species. CONCLUSIONS: According to our data, M. globosa was the most prevalent species in the skin of healthy individuals which recovered only in the yeast form. However, the Mycelial form of M. globosa was isolated as the dominant species from pityriasis versicolor lesions. Therefore, the role of predisposing factors in the conversion of this yeast to mycelium and its subsequent involvement in pityriasis versicolor pathogenicity should be considered. PMID- 15119959 TI - Serum TNF-alpha in psoriasis after treatment with propylthiouracil, an antithyroid thioureylene. AB - BACKGROUND: Tumor necrosis factor-alpha (TNF-alpha) and its receptors play important roles in the development and persistence of psoriatic plaques. The antithyroid thioureylenes, propylthiouracil and methimazole, are effective in the treatment of patients with psoriasis with a significant number of patients showing clearing or near clearing of their lesions after a several weeks of treatment. METHODS: The present study examined the effect of treatment with propylthiouracil, given in a dose of 100 mg every 8 hours for 3 months, on the serum levels of TNF-alpha in 9 patients with plaque psoriasis. RESULTS: Propylthiouracil therapy did not result in a significant decline in serum TNF alpha concentrations. CONCLUSIONS: The findings suggest that the therapeutic effect of propylthiouracil in psoriasis appears not to be related to any change in the concentration of TNF-alpha but occurs via an anti-proliferative mechanism as we have previously speculated. PMID- 15119960 TI - Antimicrobial activity of copper surfaces against suspensions of Salmonella enterica and Campylobacter jejuni. AB - BACKGROUND: Salmonella enterica and Campylobacter jejuni are amongst the more prevalent bacterial pathogens that cause foodborne diseases. These microorganisms are common contaminants of poultry and poultry products. This study was aimed to evaluate the antibacterial activity of metallic copper surfaces on these important enteropathogens, and to determine the potential acquisition of copper by food exposed to this metal. RESULTS: The antibacterial activity of copper surfaces was evaluated overlying them with suspensions of 10(6) CFU/ml of S. enterica and C. jejuni. Bacterial counts obtained after 0, 2, 4 and 8 hours at 10 degrees C and 25 degrees C were compared with those obtained in stainless steel and a synthetic polymer as control surfaces. The results showed that when these enteropathogens were kept in contact with copper a significant antibacterial activity was noted, on the contrary when the same load of pathogen suspensions were tested over the control surfaces it was found that the bacterial counts remained unchanged or even increased with time. The potential acquisition of copper by food exposed to this surface was also evaluated. Meat exposed for one hour to a copper surface adsorbed residual copper in a time dependent manner. CONCLUSIONS: These results shows that metallic copper surfaces have an antibacterial activity against S. enterica and C. jejuni and suggest its potential application as an inhibitory agent in the various stages of the food processing operations. PMID- 15119961 TI - Systematic analysis of T7 RNA polymerase based in vitro linear RNA amplification for use in microarray experiments. AB - BACKGROUND: The requirement of a large amount of high-quality RNA is a major limiting factor for microarray experiments using biopsies. An average microarray experiment requires 10-100 microg of RNA. However, due to their small size, most biopsies do not yield this amount. Several different approaches for RNA amplification in vitro have been described and applied for microarray studies. In most of these, systematic analyses of the potential bias introduced by the enzymatic modifications are lacking. RESULTS: We examined the sources of error introduced by the T7 RNA polymerase based RNA amplification method through hybridisation studies on microarrays and performed statistical analysis of the parameters that need to be evaluated prior to routine laboratory use. The results demonstrate that amplification of the RNA has no systematic influence on the outcome of the microarray experiment. Although variations in differential expression between amplified and total RNA hybridisations can be observed, RNA amplification is reproducible, and there is no evidence that it introduces a large systematic bias. CONCLUSIONS: Our results underline the utility of the T7 based RNA amplification for use in microarray experiments provided that all samples under study are equally treated. PMID- 15119962 TI - Assembly of the giant protein projectin during myofibrillogenesis in Drosophila indirect flight muscles. AB - BACKGROUND: Projectin is a giant modular protein of Drosophila muscles and a key component of the elastic connecting filaments (C-filaments), which are involved in stretch activation in insect Indirect Flight Muscles. It is comparable in its structure to titin, which has been implicated as a scaffold during vertebrate myofibrillogenesis. METHODS: We performed immunofluorescence studies on Drosophila pupal tissue squashes and isolated myofibrils to identify the pattern of appearance and assembly for projectin and several other myofibrillar proteins, using both wild type and mutant fly stocks. RESULTS AND CONCLUSIONS: In the first step of assembly, projectin immunolocalization appears as random aggregates colocalizing with alpha-actinin, kettin and Z(210), as well as, F-actin. In the second step of assembly, all these proteins become localized within discrete bands, leading ultimately to the regularly spaced I-Z-I regions of myofibrils. This assembly process is not affected in myosin heavy chain mutants, indicating that the anchoring of projectin to the thick filament is not essential for the assembly of projectin into the developing myofibrils. In the actin null mutation, KM88, the early step involving the formation of the aggregates takes place despite the absence of the thin filaments. All tested Z-band proteins including projectin are present and are colocalized over the aggregates. This supports the idea that interactions of projectin with other Z-band associated proteins are sufficient for its initial assembly into the forming myofibrils. In KM88, though, mature Z-bands never form and projectin I-Z-I localization is lost at a later stage during pupal development. In contrast, treatment of adult myofibrils with calpain, which removes the Z-bands, does not lead to the release of projectin. This suggests that after the initial assembly with the Z-bands, projectin also establishes additional anchoring points along the thick and/or thin filaments. In conclusion, during pupation the initial assembly of projectin into the developing myofibril relies on early association with Z-band proteins, but in the mature myofibrils, projectin is also held in position by interactions with the thick and/or the thin filaments. PMID- 15119963 TI - A plasmid-based system for expressing small interfering RNA libraries in mammalian cells. AB - BACKGROUND: RNA interference (RNAi) is an evolutionarily conserved process that functions to inhibit gene expression. The use of RNAi in mammals as a tool to study gene function has rapidly developed in the last couple of years since the discovery that the function-inhibiting units of RNAi are short 21-25 nt double stranded RNAs (siRNAs) derived from their longer template. The use of siRNAs allows for gene-specific knock-down without induction of the non-specific interferon response in mammalian cells. Multiple systems have been developed to introduce siRNAs into mammals. One of the most appealing of these techniques is the use of vectors containing polymerase III promoters to drive expression of hairpin siRNAs. However, there are multiple limitations to using hairpin siRNA vectors including the observation that some are unstable in bacteria and are difficult to sequence. RESULTS: To circumvent the limitation of hairpin siRNA vectors we have developed a convergent opposing siRNA expression system called pHippy. We have generated pHippy vectors or expression cassettes that knock down the expression of both reporter and endogenous genes. As a proof of principle that pHippy can be used to generate random siRNA libraries, we generated a small siRNA library against PGL3 luciferase and demonstrated that we could recover functional siRNAs that knock down PGL3 luciferase. CONCLUSIONS: siRNA is a powerful tool to study gene function. We have developed a new vector with opposing convergent promoters for the expression of siRNAs, which can be used to knock down endogenous genes in a high throughput manner or to perform functional screening with random or cDNA-derived siRNA libraries. PMID- 15119964 TI - ASAView: database and tool for solvent accessibility representation in proteins. AB - BACKGROUND: Accessible surface area (ASA) or solvent accessibility of amino acids in a protein has important implications. Knowledge of surface residues helps in locating potential candidates of active sites. Therefore, a method to quickly see the surface residues in a two dimensional model would help to immediately understand the population of amino acid residues on the surface and in the inner core of the proteins. RESULTS: ASAView is an algorithm, an application and a database of schematic representations of solvent accessibility of amino acid residues within proteins. A characteristic two-dimensional spiral plot of solvent accessibility provides a convenient graphical view of residues in terms of their exposed surface areas. In addition, sequential plots in the form of bar charts are also provided. Online plots of the proteins included in the entire Protein Data Bank (PDB), are provided for the entire protein as well as their chains separately. CONCLUSIONS: These graphical plots of solvent accessibility are likely to provide a quick view of the overall topological distribution of residues in proteins. Chain-wise computation of solvent accessibility is also provided. PMID- 15119965 TI - A structural study for the optimisation of functional motifs encoded in protein sequences. AB - BACKGROUND: A large number of PROSITE patterns select false positives and/or miss known true positives. It is possible that--at least in some cases--the weak specificity and/or sensitivity of a pattern is due to the fact that one, or maybe more, functional and/or structural key residues are not represented in the pattern. Multiple sequence alignments are commonly used to build functional sequence patterns. If residues structurally conserved in proteins sharing a function cannot be aligned in a multiple sequence alignment, they are likely to be missed in a standard pattern construction procedure. RESULTS: Here we present a new procedure aimed at improving the sensitivity and/ or specificity of poorly performing patterns. The procedure can be summarised as follows: 1. residues structurally conserved in different proteins, that are true positives for a pattern, are identified by means of a computational technique and by visual inspection. 2. the sequence positions of the structurally conserved residues falling outside the pattern are used to build extended sequence patterns. 3. the extended patterns are optimised on the SWISS-PROT database for their sensitivity and specificity. The method was applied to eight PROSITE patterns. Whenever structurally conserved residues are found in the surface region close to the pattern (seven out of eight cases), the addition of information inferred from structural analysis is shown to improve pattern selectivity and in some cases selectivity and sensitivity as well. In some of the cases considered the procedure allowed the identification of functionally interesting residues, whose biological role is also discussed. CONCLUSION: Our method can be applied to any type of functional motif or pattern (not only PROSITE ones) which is not able to select all and only the true positive hits and for which at least two true positive structures are available. The computational technique for the identification of structurally conserved residues is already available on request and will be soon accessible on our web server. The procedure is intended for the use of pattern database curators and of scientists interested in a specific protein family for which no specific or selective patterns are yet available. PMID- 15119967 TI - Using morphometry of the larval rostellar hooks to distinguish Iranian strains of Echinococcus granulosus. AB - Human cystic echinococcosis is widely distributed throughout Iran, where sheep, cattle, goats, camels and other animals act as intermediate hosts. It appears that morphometry of the rostellar hooks on the protoscoleces of the causative parasite, Echinococcus granulosus, can be used to separate the strains or variants of the parasite to be found in humans and livestock in Iran. Multivariate statistical procedures, including principal-component analysis and discriminant-function analysis (DFA), were used to explore the morphometric data obtained from the larval hooks of Iranian samples of E. granulosus. Although five physical variables were initially considered, the results of the PCA indicated that just two factors (based on the length of hooks and number of hooks) accounted for 91.6% of the variance observed in the hook measurements. The results of the DFA allowed the correct classification of all the samples. The presence in Iran of the sheep and camel strains of the parasite was confirmed. The sheep strain was the most common variant encountered, being recovered from sheep, cattle, goats, humans and occasionally camels. Most of the metacestode samples from camels and a few of those from cattle (five of 30), sheep (one of 27) and humans (two of 23) were found to be of the camel strain. These results indicate that larval-hook morphology is a valid method for identifying E. granulosus strains in Iran, and one that is potentially useful for epidemiological studies. PMID- 15119966 TI - An application of conditional logistic regression and multifactor dimensionality reduction for detecting gene-gene interactions on risk of myocardial infarction: the importance of model validation. AB - BACKGROUND: To examine interactions among the angiotensin converting enzyme (ACE) insertion/deletion, plasminogen activator inhibitor-1 (PAI-1) 4G/5G, and tissue plasminogen activator (t-PA) insertion/deletion gene polymorphisms on risk of myocardial infarction using data from 343 matched case-control pairs from the Physicians Health Study. We examined the data using both conditional logistic regression and the multifactor dimensionality reduction (MDR) method. One advantage of the MDR method is that it provides an internal prediction error for validation. We summarize our use of this internal prediction error for model validation. RESULTS: The overall results for the two methods were consistent, with both suggesting an interaction between the ACE I/D and PAI-1 4G/5G polymorphisms. However, using ten-fold cross validation, the 46% prediction error for the final MDR model was not significantly lower than that expected by chance. CONCLUSIONS: The significant interaction initially observed does not validate and may represent a type I error. As data-driven analytic methods continue to be developed and used to examine complex genetic interactions, it will become increasingly important to stress model validation in order to ensure that significant effects represent true relationships rather than chance findings. PMID- 15119968 TI - Schistosomes in the Xe Kong river of Cambodia: the detection of Schistosoma mekongi in a natural population of snails and observations on the intermediate host's distribution. AB - A natural population of Neotricula aperta (Gastropoda: Pomatiopsidae) in the Xe Kong river of Cambodia was found to be infected with Schistosoma mekongi- apparently the first time this parasite has been found in snails outside the Mekong river. Only 0.14% of the N. aperta collected were found infected. Potential habitats for N. aperta were examined in selected rivers of central and southern Laos and Cambodia, so that the density of N. aperta at each site could be estimated. At survey sites where sufficient snails were collected, the population was also screened for infection with S. mekongi. The geographical distribution of N. aperta outside the Mekong river is shown to be much greater than previously documented; this is in agreement with predictions based on palaeo geographical models and the evolutionary history and historical biogeography (phylogeography) of the species. The Xe Kong river is identified as a potential source of N. aperta colonists entering Cambodia from Laos. The findings, which indicate that there is a risk of human infection with S. mekongi in areas distant from the Mekong river, have important implications for schistosomiasis-control strategies in the region. PMID- 15119969 TI - An investigation of persistent microfilaridermias despite multiple treatments with ivermectin, in two onchocerciasis-endemic foci in Ghana. AB - If ivermectin-based programmes for the control of human onchocerciasis are to be successful, the drug must remain effective for as long as necessary. In an open, case-control study, an attempt was made to determine if the persistent, significant, Onchocerca volvulus microfilaridermias seen in some individuals who had received at least nine treatments with ivermectin were the result of the development of drug resistance in the parasite. Twenty-one of these 'sub-optimal' responders (cases) were matched, by age, weight, number of treatments, locality and skin microfilarial counts, with seven amicrofilaridermic responders and 14 ivermectin-naive subjects. The number of treatments taken, any potential drug interactions and significant underlying disease were determined from detailed clinical and laboratory studies. Each subject was treated with ivermectin during the study, so that plasma concentrations of the drug could be determined for 72 h from the time of dosage. The microfilarial and adult-worm responses to this treatment were assessed from skin microfilarial counts (obtained before the treatment and at days 8, 90 and 365 post-treatment), day-90 embryogrammes, and the results of fly-feeding experiments. Parasite-sensitivity criteria for various time-points were derived from earlier data on skin microfilaridermias and the effects of ivermectin on the adult worms. The results indicate that the significant microfilaridermias that persist despite multiple treatments with ivermectin are mainly attributable to the non-response of the adult female worms and not to inadequate drug exposure or other factors. The possibility that some adult female worms have developed resistance to ivermectin cannot be excluded. These results justify the routine monitoring of treatment efficacy in any ivermectin-based programme of disease control. PMID- 15119970 TI - The infectivity and antigenicity of Toxocara canis eggs can be retained after long-term preservation. AB - Suspensions of fertilized eggs of Toxocara canis were mixed with 2% neutral formalin and preserved at 4 degrees C. When, after storage for 0, 12, 18, 21 and 24 months, samples of the eggs were incubated at 30 degrees C for 12 days, 96.8%, 92.6%, 74.1%, 51.0% and 19.3% of the eggs in the samples were found to embryonate. The embryonated eggs produced from the fertilized eggs preserved (in 2% neutral formalin at 4 degrees C) for 0, 12, 18 and 21 months were then tested for their infectivity to BALB/c mice, each mouse being given 800 embryonated eggs. The numbers of larvae recovered from the mice and the sites from which they were recovered, 2 or 14 days post-infection, appeared unaffected by the length of storage of the eggs. The infected mice all had similar eosinophil counts in their peripheral blood and similar serum titres of Toxocara-specific IgM and IgG antibodies, and cultures of their spleen cells produced similar amounts of interleukin-4, interleukin-5 and interferon-gamma when stimulated with concanavalin A. The results of SDS-PAGE indicated that egg preservation for at least 21 months had no effect on the excretory-secretory antigens in samples of medium from cultures of infective larvae released from the eggs. In summary, at least 50% of the fertilized eggs preserved in 2% neutral formalin at 4 degrees C for 21 months could fully embryonate and then had the same infectivity and antigenicity as embryonated fresh eggs. PMID- 15119972 TI - High mortality among patients with bacterial meningitis in a rural hospital in Tanzania. AB - Although the disease is an important cause of mortality in the region, most published reports on bacterial meningitis in East Africa are from urban referral hospitals. Poor laboratory facilities make diagnosis difficult in the area and treatment is limited to inexpensive antibiotics. The case-fatality 'rate' in one rural hospital in Tanzania, the Ndala Mission Hospital (NMH), appears to have increased dramatically over recent years, perhaps as the result of increasing resistance to ampicillin and chloramphenicol. The aim of the present study, which was partially retrospective and partially prospective, was to review the number, characteristics and outcome of children admitted to this hospital with bacterial meningitis and to investigate possible resistance of the causative micro organisms to the antibiotics used. Data from the 181 children who were admitted with bacterial meningitis [confirmed by the examination of Gram-stained smears of cerebrospinal fluid (CSF)] between 1999 and 2002 were retrospectively reviewed. The overall mortality among these children was 51%. No seasonal pattern was observed in the number of cases. In a 2-month prospective study in 2002, CSF samples from 19 consecutive cases were collected in Trans-Isolate medium and shipped to the Academic Medical Center in Amsterdam for culture and analysis of antibiotic susceptibility. For only eight (42%) of the cases was there agreement between the species of bacterium identified, by Gram-staining, in Tanzania and that identified, by culture, in The Netherlands. As there was no evidence of resistance to ampicillin and the antibiotics used in the NMH were found to be of good quality, the cause of the high mortality in the NMH remains uncertain. Poor laboratory testing, long doctor-patient delays and/or poor drug administration on the wards may all be contributory factors. Attempts will now be made to address each of these problems. PMID- 15119971 TI - Strongyloidiasis: challenges in diagnosis and management in non-endemic Kuwait. AB - Among immunocompromised individuals, hyper-infection with Strongyloides stercoralis may occur and lead to fatal strongyloidiasis. To clinicians and laboratory diagnosticians in non-endemic countries such as Kuwait, this severe infection poses a particular problem. The clinical histories and signs and symptoms of four Kuwaiti cases of S. stercoralis hyper-infection were reviewed. Each of the four was found not only to have lived in an area where S. stercoralis was endemic but also to have been treated with immunosuppressive steroids (for medical problems unrelated to the nematode infection). When they presented with undiagnosed hyper-infections their clinical features were confusing. Three of the cases, all with low eosinophil counts, died but the other, who was treated with thiabendazole, survived. In the light of these observations, healthy medical examinees who had recently moved from endemic zones were checked for asymptomatic S. stercoralis infection, both by stool examination and ELISA-based serology. Of 381 stool samples investigated over a 3-month period, 183 (48%) were found positive for helminths, 7% for S. stercoralis. Of 198 individuals from endemic zones who were screened after another medical examination, 71 (35.8%) were found positive for intestinal helminth parasites, including one (1.45%) infected with S. stercoralis. Although ELISA appear reliable in making a presumptive diagnosis of strongylodiasis, the results of such assays are not very specific and are best interpreted in conjunction with the patient's clinical status. The concurrent administration of anthelminthics to patients prescribed steroids who, because they live or have lived in an area where S. stercoralis is endemic, are at risk of infection with the nematode, should be considered. PMID- 15119973 TI - Use of CHROMagar medium in the differentiation of Candida species: is it cost effective in developing countries? AB - The opportunistic infection of humans with Candida is becoming more common. As several species of Candida are relatively insusceptible to the commonly used antifungal drugs, rapid identification of the species involved can facilitate effective treatment. CHROMagar Candida medium (CCM) is a commercial product designed to allow the rapid identification of Candida to species level. To explore the potential usefulness of CCM in a developing country, attempts were made to identify the Candida species in 107 Indian isolates (obtained, consecutively, from 90 clinical specimens, over a year-long period, in a tertiary level teaching hospital), using CCM and more conventional methods in parallel. The most common species appeared to be C. tropicalis (representing 40% of the isolates), followed by C. albicans (28%) and C. glabrata (23%). Although use of CCM allowed the isolates from 84 (93.3%) of the clinical specimens to identified to species level within 48 h, it took at least 7 days to identify the yeasts in 90% of the specimens using the more conventional procedures. With the results of the conventional methods set as the 'gold standard', the use of CCM appeared to allow all of the C. albicans, C. tropicalis and C. krusei isolates and most (92%) of the C. glabrata to be correctly identified. The costs/isolate identified with the CCM were no more than those of the conventional methods. As many (67%) of the isolates examined were of potentially drug-resistant yeasts (C. tropicalis, C. glabrata or C. krusei), there is clearly a need to identify local isolates quickly, to prevent treatment with ineffective drugs. In terms of both performance and cost, CCM appears to be a good method to use. PMID- 15119974 TI - The distribution and ecological 'preferences' of the tick Amblyomma cajennense (Acari: Ixodidae), an ectoparasite of humans and other mammals in the Americas. AB - The distribution and ecological 'preferences', in the Nearctic and Neotropical regions, of the prominent, pathogen-carrying tick Amblyomma cajennense (Fabricius) were reviewed, on the basis of the published literature, satellite imagery, and the examination of several tick collections. The tick has been collected from the southern U.S.A. (Texas and Florida) to northern Argentina, but is probably absent from Uruguay and southern Brazil (the few records of the species in these areas probably represent mis-identifications). The ecological conditions prevailing in the areas where the tick has been reported were characterized using remotely sensed data on temperature and vegetation stress (as indicated by the normalized derived vegetation index, or NVDI). In Mexico and the U.S.A., the species is found in areas where the mean temperature is around 13-16 degrees C and the NVDI is high. South of Venezuela, however, the tick is only common in zones where the mean temperature is 18-20 degrees C and the NVDI is again high. It is relatively low mean temperatures and differences in the seasonal patterns of rainfall that seem to limit the tick's colonization of areas to the north of its current distribution. Low temperatures also seem to be keeping the tick out of mountainous areas, such as the Sierra Madre in Mexico and the pre-Andean hills in Argentina. The southern distribution of A. cajennense is mainly restricted by relatively low temperatures and not by low humidity. PMID- 15119975 TI - High frequency of the genetic polymorphisms associated with sulfadoxine pyrimethamine resistance, among Plasmodium falciparum isolates from Sao Tome and Principe, West Africa. PMID- 15119976 TI - The cerebral-malaria-associated expression of RANTES, CCR3 and CCR5 in post mortem tissue samples. PMID- 15119977 TI - Epidemiology of dengue fever in Trinidad, West Indies: the outbreak of 1998. PMID- 15119978 TI - Matrix metalloproteinases: contribution to pathogenesis, diagnosis, surveillance and treatment of abdominal aortic aneurysms. AB - BACKGROUND: Aortic abdominal aneurysm (AAA) represents a common chronic degenerative disease of the aortic wall. Chronic inflammation and enzymatic degradation of elastic lamellae and extracellular matrix (ECM) proteins constitute the most prominent characteristics of AAAs. There is mounting evidence that matrix metalloproteinases (MMPs) are the predominant proteinases in the AAA wall. These enzymes represent a potential target for therapeutic intervention to modify vascular pathology. This paper is an overview of matrix metalloproteinases and their role in the pathophysiology, diagnosis and treatment of AAA. LITERATURE SEARCH: Comprehensive search of the MEDLINE, EMBASE and HEAL-Link databases from 1980 to 2003. FINDINGS: Increased levels of MMPs expression and activity have been demonstrated within the aortic wall of AAA, associating with histological alterations. An imbalance between MMPs and their inhibitors (Tissue Inhibitors of Matrix Metalloproteinases - TIMPs), may tip the equilibrium towards matrix degradation. MMPs as systemic biochemical markers of AAAs may contribute to diagnosis of unsuspected AAAs or to the surveillance of patients with small AAAs. Evidence of variations in MMPs, TIMPs and their mediator genes promoting the increased inheritance susceptibility of AAAs is less well documented. However,a broad spectrum of pharmaceutical agents (e.g. doxycycline, statins etc.) is known to inhibit MMP activity and attenuate medial destruction. CONCLUSION: Randomized clinical studies in patients in the early stages of AAA or in healthy individuals with great propensity to AAA development are required to demonstrate the causative relationship between MMPs and AAA. It still remains obscure whether long-term administration of MMP inhibitors can decelerate or even prevent the need for surgical repair. PMID- 15119979 TI - Efficacy of risedronate on clinical vertebral fractures within six months. AB - OBJECTIVE: Postmenopausal osteoporotic women with pre-existing or new incident vertebral fractures are at high risk for future fracture, so prompt treatment is warranted. Risedronate has been shown to reduce the incidence of radiographically defined vertebral fractures by approximately two-thirds within 1 year. RESEARCH DESIGN: This study examined the effects of risedronate treatment on the time course of the reduction in the risk of clinical vertebral fractures (i.e., symptomatic fractures), on the risk of moderate-to-severe radiographic vertebral fractures, and on height. RESULTS: In 2442 postmenopausal women with prevalent vertebral fractures from the Vertebral Efficacy with Risedronate Therapy (VERT) studies who received either risedronate 5 mg or placebo, daily risedronate reduced the risk of clinical vertebral fractures within 6 months (RR = 0.08, 95% CI 0.01-0.63), and by 69% at 1 year (RR = 0.31, 95% CI 0.12, 0.78). At 1 year, risedronate also reduced the risk of moderate-to-severe radiographically-defined vertebral fractures by 71% (RR = 0.29 95% CI 0.16, 0.54). Height loss was attenuated with treatment, most notably in patients who experienced new vertebral fractures, with a median difference of 0.73 cm compared with subjects receiving placebo (p = 0.005). CONCLUSION: Risedronate reduces the risk of clinical vertebral fractures in postmenopausal women with osteoporosis within 6 months of commencing treatment. PMID- 15119980 TI - Comparison of temazepam 7.5 mg with temazepam 15 mg for the treatment of transient insomnia. AB - OBJECTIVE: To demonstrate the equivalent efficacy of temazepam 7.5 mg and temazepam 15 mg for the treatment of transient insomnia. RESEARCH DESIGN AND METHODS: This was a double-blind, parallel group, multicenter study. Healthy male and female subjects with previous but not current complaints of transient insomnia were enrolled. Transient insomnia was induced in the sleep laboratory by means of the 'first night' effect and by implementing a 2-h phase advance. The effects of both doses of temazepam on polysomnographic (PSG) measures of sleep were evaluated for one night. Latency to persistent sleep (LPS) and total sleep time (TST) were designated as the primary efficacy endpoints. RESULTS: One hundred and thirty-one subjects completed the study: 65 received the 7.5-mg dose, and 66 received the 15-mg dose. Treatment groups begin with the lowest effective dose, i.e., 7.5 mg. were well matched based on background demographics. No statistically significant differences between doses were detected for LPS, TST,or any other objective (PSG) measure of sleep. Furthermore, both doses were found to be clinically equivalent for LPS and TST based on predetermined criteria. Temazepam was well tolerated, and no significant differences between doses were found for adverse event (AE) incidence, mean score on the Digit Symbol Substitution Task, or mean scores on questions related to tolerability from the Leeds Sleep Evaluation Questionnaire. CONCLUSIONS: The 7.5-mg and 15-mg doses of temazepam were equally effective for the treatment of transient insomnia. In keeping with current practice guidelines, initiation of treatment with temazepam for transient insomnia should PMID- 15119981 TI - Comment on: comparison of 5 days of ER clarithromycin versus 10 days of penicillin V for the treatment of streptococcal pharyngitis/tonsillitis. PMID- 15119982 TI - Generic replacement of clozapine: a simple decision model from a Canadian perspective. AB - OBJECTIVE: Increased incidence of relapse has been differences in relapse incidence. The difference at reported upon switching patients with schizophrenia from brand name to generic clozapine. The cost of treating relapsed patients could offset the reduced drug acquisition cost associated with switching. A decision model was designed to predict the relapse incidence at which switching to generic clozapine is cost-neutral. RESEARCH DESIGN AND METHODS: A hypothetical cohort of 100 patients with schizophrenia stabilized on brand name clozapine was considered either to remain on the brand name product, or to switch to the generic version. Medication effectiveness data were taken from two reports following patients who underwent generic replacement of clozapine. Direct costs associated with each treatment were projected from a Canadian Ministry of Health perspective, considering drug acquisition and treatment of relapse. Unit costs were derived from published sources. MAIN OUTCOME MEASURES: Direct costs of the two treatment regimens were compared based on which switching to generic clozapine would result in no direct cost saving was determined. RESULTS: Switching a patient to generic clozapine would save 1241 Canadian dollars annually if the patient did not relapse, and would cost 9823 Canadian dollars if the patient relapsed. Assuming an 11% difference in relapse for patients taking brand name and generic clozapine, respectively, switching 100 patients to generic clozapine would save 24 Canadian dollars per patient. If the relapse difference for patients taking generic clozapine is 28%, the switch to the generic medication would cost 1857 Canadian dollars per patient. Switching patients from brand name clozapine to generic clozapine was predicted to be neutral to direct costs when the absolute difference in relapse incidence was 11.2%. CONCLUSIONS: Switching to a generic medication may not always reduce direct costs. Physicians, patients and third party payers should consider the potential consequences before instituting generic replacement of clozapine for economic reasons. PMID- 15119983 TI - Program savings associated with switching testosterone intramuscular injections to topical gel in HIV infected males. AB - BACKGROUND: HIV-1 infected male patients commonly require testosterone supplementation and intramuscular testosterone (IM) is the most frequently used supplementation method. Currently, the economic burden of testosterone supplementation is borne to a large degree by the clinic providing services to the HIV infected male. Increasingly, clinics are seeking alternative methods to provide supplementation. This analysis describes the economic trade-offs associated with the utilization of topically applied testosterone supplement relative to current supplementation techniques within the clinic setting. METHODS: Twenty-three patients that had been receiving IM were switched to topical testosterone gel (TOP). A 1-month time-and-motion study, identification of IM administration specific supplies, and costs for acquisition and disposal of these supplies were conducted or gathered to quantify costs of providing bi weekly IM to patients. The analysis describes potential resource savings by switching 23 HIV patients served in a safety-net provider setting from IM to TOP. The analysis considers both a pre and post-evaluation period whereby patients received IM in the pre-evaluation period and then were switched to TOP in the follow-up period. Economic assessments considered direct medical costs for the preparation and administration of the product but did not incorporate other outcomes in the analyses. RESULTS: Only one of 23 patients switched experienced recurrence of symptoms and no patients developed adverse effects associated with the switch. In contrast, substantial resources could be realized with the utilization of the topically applied testosterone relative to the injectable formulations with approximately $80,938 fewer dollars spent on provision of IM. CONCLUSIONS: Switching patients from IM to TOP resulted in no deleterious affects and is estimated to have saved the clinic considerable resources, even if TOP had to be provided by the clinic. PMID- 15119984 TI - Drug costs and bacterial susceptibility after implementing a single fluoroquinolone use policy at a university hospital. AB - The University of Kentucky Hospital investigated the feasibility of choosing a sole fluoroquinolone for its formulary in an effort to reduce costs without affecting clinical outcomes. A three-step process was used to plan, implement, and monitor the selection program. Based on the range of clinical indications, safety profile, local susceptibility, cost, and dosing convenience, levofloxacin was chosen over ciprofloxacin and gatifloxacin as the sole fluoroquinolone. Since the implementation of the program in May 2001, susceptibility to levofloxacin has been maintained or increased for the most common pathogens. In addition, University Hospital has saved nearly 100,000 dollars in antibiotic acquisition costs during the first 12 months after the switch. This assessment did not take into account effects in clinical outcomes, such as clinical failures (such as readmission rates), mortality, and adverse events, or measure changes in overall medical expenditures beyond drug acquisition costs. In the future, monitoring of overall patient care and medical care costs, in addition to susceptibility patterns and drug costs, will allow for a better understanding of the long-term benefits of this switch. PMID- 15119985 TI - Quality assessment of meta-analyses of RCTs of pharmacotherapy in major depressive disorder. AB - BACKGROUND: Meta-analyses (MAs) of randomized controlled trials (RCTs) have the potential to provide the highest level of evidence, but the quality of published MAs has not been systematically assessed. Therefore, we determined reliability was significant (kappa = 0.89; p < 0.05). the quality of reporting in MAs of RCTs of pharmacotherapy for major depressive disorder (MDD) in adults (18-65 years) without comorbidities and examine trends over time. METHODS: MEDLINE, EMBASE, Healthstar, Psychlit and Cochrane databases were searched (1980-2002) by 4 independent reviewers for MAs of RCTs. Articles meeting inclusion criteria were blinded. Inter-rater reliability (kappa) was evaluated using a test-retest strategy on 4 articles. Quality was (p = 0.74) did not detect a difference in quality of assessed using the QUOROM checklist. Time trends were evaluated by calculating Spearman's rho. RESULTS: One hundred articles were identified, 68 were excluded [co-morbidities (9), inappropriate comparator (13), inappropriate outcome (15), article not available (5), inappropriate patient population (15), and inappropriate study design (11)]; 32 were included. Initial kappa was 0.81 (p < 0.05). After resolution of disagreements, the test-retest The mean overall quality score was 50.2% (SD 15.8%, range = 16.7-88.9%). The overall score for Titles was very poor (22%), Abstracts (40%) and Methods (49%) were poor, while overall Results score was minimally acceptable (54%). Good quality scores were found for Introduction (91%) and Discussion (97%). No time trends were identified using Spearman's correlation analysis (rho 0.05; p = 0.79). The Mann-Whitney U test articles published before and after the QUOROM. CONCLUSION: Despite quality guidelines, the average quality of published MAs of antidepressants is barely acceptable (50.2%). A need exists for adherence to standardized reporting and quality guidelines. PMID- 15119986 TI - Efficacy, safety and cost-effectiveness of Amphotericin B Lipid Complex (ABLC): a review of the literature. AB - Amphotericin B Lipid Complex (ABLC) was the first lipid-based formulation of amphotericin B (AmB) to be developed, it was designed to provide a less toxic alternative to conventional AmB without compromising efficacy. Preclinical and early clinical data relating to ABLC have been presented in previous reviews. This paper reviews more recent published data on the efficacy, safety and cost effectiveness of ABLC. All published manuscripts and conference abstracts were searched on MEDLINE, BIOL and SCIN for the period between January 1997 and August 2003. Comparative and non-comparative studies of ABLC are usually mild or moderate and are manageable were considered. Comparative studies and additional data from non-comparative studies suggest that ABLC 5 mg/kg/day is safe and effective for the treatment of documented or suspected systemic fungal infections in adults and children who are refractory to or intolerant of conventional AmB. ABLC is effective against a wide range of pathogens and efficacy is at least as good as conventional AmB or the other lipid-based formulations. The safety profile of ABLC is improved compared with conventional AmB; ABLC is less nephrotoxic than conventional AmB and can be given safely to patients with pre existing renal impairment. The most commonly reported adverse effects are transient infusion-related events, including chills, fever, nausea and vomiting, which with premedication. Comparative studies suggest that ABLC is a cost effective treatment option compared with conventional AmB or other lipid-based formulations of amphotericin B. PMID- 15119987 TI - Soccer, neurotrauma and amyotrophic lateral sclerosis: is there a connection? AB - Trauma has long been hypothesized but never proven to be a risk factor for amyotrophic lateral sclerosis (ALS). This hypothesis may now have a renaissance due to recent reports in the lay press on 'the Italian motoneuron mystery', i.e. the disclosure of 33 diagnosed ALS cases in a subpopulation of 24000 soccer players of the top three Italian divisions from the 1960s to 1996. Could the repetitive brain trauma that soccer players experience for controlling and advancing the ball with their heads represent an environmental risk factor for developing ALS in genetically predisposed individuals? By critically reviewing the scarce literature and 'surrounding evidence' (Medline, CDC, lay press, Italian health officials), we have looked for a potential relationship between (1) soccer and head trauma and (2) head trauma and subsequent development of amyotrophic lateral sclerosis. Whereas the brain traumatizing effect of soccer seems to be out of the question, the findings of the few retrospective studies on ALS and neurotrauma are conflicting. Taken together, however, the literature would still support the concept of soccer, head trauma, and ALS being interrelated, with high levels of athleticism/physical activity perhaps playing an additive part. To further clarify this issue, extensive prospective epidemiological investigations on ALS following neurotrauma as well as carefully designed animal studies will have to be conducted. PMID- 15119988 TI - The road to efficient and effective stuttering management: information for physicians. AB - Physicians are often the first point of contact when children's speech begins to be disrupted by stuttering behaviors such as sound repetitions and prolongations. For this reason, we feel it is important that they are accurately informed with regards to the nature of stuttering and the available treatment options before making referrals to speech-language pathologists. Stuttering is by definition, an involuntary disorder and remains that way throughout life. Its cause is still unknown and the only true form of remission appears to be the natural, spontaneous recovery that occurs in up to 80% of those children afflicted. No therapeutic course has seemed to change this figure and the prevalence of stuttering in the general population has remained stable, suggesting that speech therapy has never 'cured' stuttering. Therefore, we suggest that therapeutic intervention for stuttering should be best directed towards 'efficient' and 'effective' symptom reduction. Until recently, intervention options for children and adults who stutter have generally been limited consist of countless hours of speech retraining (teaching people 'how to talk again'), while attempting to bring the disorder under voluntary control. The common end-results of these procedures include unnatural speech patterns that are difficult to maintain in all situations and highly prone to relapse, thus, reinforcing the notion that stuttering is highly resistant to treatment. However, miniaturized digital technology now allows those who stutter to take advantage of auditory effects that 'inhibit' stuttering. 'Choral speech' or speaking in unison has long been known to make those who stutter immediately fluent without compromising speech naturalness. All in-the-ear devices can emulate choral speech effects by altering auditory feedback. Therapeutic protocols using these devices can be quickly and efficiently implemented. Furthermore, they are showing high levels of long-term effectiveness with regards to reducing stuttering frequency and maintaining speech naturalness. PMID- 15119989 TI - Examination of nighttime sleep-related problems during double-blind, placebo controlled trials of galantamine in patients with Alzheimer's disease. AB - BACKGROUND: Acetylcholinesterase inhibitors (AChEI) are now widely used as treatment for Alzheimer's disease (AD). Their cholinomimetic action has the potential to influence sleep quality and donepezil has been associated with sleeprelated adverse events. This study examined whether galantamine, an AChEI with nicotinic modulation, is associated with nighttime sleeprelated problems. METHODS: Using data combined from three randomized, double-blind trials, galantamine (GAL) and placebo-treated patients were compared on sleep-related adverse events and concomitant medications. Verbatim descriptions by patients, as recorded by physicians, were used to classify adverse events into insomnia/sleep problems and nightmares/dreams. New concomitant medications findings from double blind, placebo-controlled were counted when prescribed for sleep-related indications. Treatment arms were 705 subjects on GAL 24 mg (12 mg BID), 279 on GAL 16 mg (8 mg BID), and 714 on placebo. RESULTS: Comparing GAL 24, GAL 16, and placebo arms, the respective rates for insomnia/sleep problems were 2.6, 1.1, and 2.2% and for nightmares were 1.1, 0.4, and 0.1%. Comparisons between GAL arms and placebo were not significant, with the exception of GAL 24 for nightmares (p = 0.02). However, due to the low frequency of nightmares in each group the clinical relevance of this difference is unknown. Rates of use for all concomitant medications indicated for sleep were 5.4, 2.9, and 4.6%, respectively, with no significant differences between GAL arms and placebo. CONCLUSION: These results support previous studies that have shown maintenance of good sleep hygiene during treatment with galantamine. PMID- 15119990 TI - Putting evidence-based medicine into clinical practice: comparing anti-resorptive agents for the treatment of osteoporosis. AB - OBJECTIVE: To compare the effectiveness of antiresorptive agents in reducing the risk of vertebral and non-vertebral fractures using data from published meta analyses and the technique of adjusted indirect comparisons. RESEARCH DESIGN AND METHODS: Pairs of agents were compared by adjusted indirect comparison of 0.56 [0.40, 0.78], respectively) in reducing the their effects relative to a common comparator (placebo) using meta-analyses published by The Osteoporosis Methodology Group and The Osteoporosis Research Advisory Group. RESULTS: Adjusted indirect comparisons identified only one pair of agents that had significantly different effects on vertebral fracture incidence: alendronate was 34% more effective than calcitonin (Relative Risk: 0.66, 95% Confidence Interval: 0.48 0.90). Alendronate was significantly more effective than risedronate, calcitonin, estrogen, etidronate, and raloxifene (Relative Risks: 0.70 [0.49, 0.99], 0.64 [0.42, 0.98], 0.59 [0.41, 0.84], 0.52 [0.32, 0.82], and incidence of non vertebral fractures. No other significant pairwise differences were observed. CONCLUSIONS: The results suggest that there are differences in anti-fracture efficacy among antiresorptive agents, particularly for non-vertebral fractures. Direct head-to-head comparisons would be needed to confirm these findings but are unlikely to be conducted. PMID- 15119991 TI - Could prolactin receptor gene polymorphism play a role in pathogenesis of breast carcinoma? AB - OBJECTIVE: Constitutive activation of various hormone and growth factor receptors is newly recognised as a common cause of tumour development. This study investigated the presence of any mutation or polymorphism of prolactin receptor (PRLR) in 38 patients with breast cancer. RESEARCH METHODS: Genomic DNA was extracted and PCR amplification was carried out for exon 1-10 of PRLR from tumoral and adjacent non-cancerous breast tissue of tumour specimens from 38 breast cancer patients. PCR products were analysed by SSCP and automatic sequencing for mutations. RESULTS: For the first time, A150C (Leu-->Ile) transversion at exon 6 of PRLR in tumour tissues, in adjacent non-cancerous breast tissues, and in blood samples of two (5.3%) out of 38 patients with breast cancer were detected. In contrast to this finding, no polymorphism of PRLR in blood samples of 100 normal individuals were found. CONCLUSION: Polymorphism of prolactin receptors might play a role in mammary carcinogenesis as a consequence of intracellular changes of PRLR signalling. PMID- 15119992 TI - Transitioning pharmacologic therapy from oral agents to insulin for type 2 diabetes. AB - Prospective intervention trials using an intensive therapy approach in patients with either type 1 or type 2 diabetes have provided evidence that achieving tight glycemic control can impede the development and progression of microvascular complications. Treatment of type 2 diabetic patients has revolved around the use of oral agents to improve insulin secretion or tissue sensitization. As therapy turns to the inclusion of insulin - most often in combination with oral agents - the physician and patient must both be prepared for the issues regarding management of an injectable agent. The objective of this article is to review current evidence supporting the benefits of adding insulin therapy to existing oral hypoglycemic regimens of patients for whom these therapies are no longer providing adequate glycemic control. Approaches to initiating insulin therapy and adjusting treatment regimens are discussed, with a view towards making the addition or switch to insulin a simple and achievable next step in treatment. PMID- 15119993 TI - Efficacy of 750-mg, 5-day levofloxacin in the treatment of community-acquired pneumonia caused by atypical pathogens. AB - BACKGROUND: Current recommended durations for treatment of atypical community acquired pneumonia (CAP) range from 10 to 21 days. However, antibiotics such as the fluoroquinolones may allow for effective, short-course regimens. OBJECTIVE: This study evaluated the efficacy of 750 mg levofloxacin for 5 days compared to a 500-mg, 10-day levofloxacin regimen for the treatment of atypical CAP. METHODS: A randomized, active-controlled, double-blind, multicenter study was conducted within the United States. Of the 528 patients enrolled in the study, 149 were diagnosed with CAP due to Legionella pneumophila, Chlamydia pneumoniae, or Mycoplasma pneumoniae. Patients' baseline symptoms were re-evaluated on Day 3 of therapy. Clinical efficacy and resolution of CAP symptoms were evaluated at the posttherapy visit (7-14 days after the last dose of active drug). RESULTS: This report represents a subgroup analysis of a previous clinical study. Among the 123 clinically evaluable patients diagnosed with atypical CAP (26 patients were unevaluable), the clinical success rates were 95.5% (63 of 66 patients) for the 750-mg group and 96.5% (55 of 57 patients) for the 500-mg group (95% CI for success rate of the 500-mg group minus that of the 750-mg group, -6.8 to 8.8). At the poststudy evaluation (31-38 days after treatment began), relapse occurred in 0.77, p < 0.001), but not correlated with RR. Lung doses initially increased from Vt = 25 to 50 mL (Nebuchamber, Aerochamber) or to 100 mL (Babyhaler) and then decreased, with increasing Vt and RR (R: -0.98 to -0.82, p < 0.001). Lung doses of fluticasone were 1.5-6-fold higher compared with budesonide, irrespective of spacer type (p < 0.001). MMAD decreased with increasing Vt and RR. Dose to the lungs of particles <2.1 microm was independent of Vt and RR. Lung dose decreases with increasing inspiratory flow (increasing Vt or RR) by increasing impaction of coarse particles in the upper airways. Deposition of particles <2.1 microm is relatively flow independent. When electrostatic charge of spacers is reduced, lung dose is pMDI dependent and spacer independent. PMID- 15120014 TI - Residual gravimetric method to measure nebulizer output. AB - The aim of this study was to assess a residual gravimetric method based on weighing dry filters to measure the aerosol output of nebulizers. This residual gravimetric method was compared to assay methods based on spectrophotometric measurement of terbutaline (Bricanyl, Astra Zeneca, France), high-performance liquid chromatography (HPLC) measurement of tobramycin (Tobi, Chiron, U.S.A.), and electrochemical measurements of NaF (as defined by the European standard). Two breath-enhanced jet nebulizers, one standard jet nebulizer, and one ultrasonic nebulizer were tested. Output produced by the residual gravimetric method was calculated by weighing the filters both before and after aerosol collection and by filter drying corrected by the proportion of drug contained in total solute mass. Output produced by the electrochemical, spectrophotometric, and HPLC methods was determined after assaying the drug extraction filter. The results demonstrated a strong correlation between the residual gravimetric method (x axis) and assay methods (y axis) in terms of drug mass output (y = 1.00 x 0.02, r(2) = 0.99, n = 27). We conclude that a residual gravimetric method based on dry filters, when validated for a particular agent, is an accurate way of measuring aerosol output. PMID- 15120015 TI - Stochastic model of particle clearance in human bronchial airways. AB - A stochastic bronchial clearance model, based on a stochastic morphometric model of the human bronchial tree, has been developed, which simulates the combined action of fast and slow bronchial clearance mechanisms by Monte Carlo methods. To model fast bronchial clearance, mucus velocities in individual airways were based on a correlation between mucus velocity and airway diameter, considering conservation of mucus flow. In addition, mucus transport was assumed to be delayed at bronchial bifurcation zones. The size dependence of the slow bronchial clearance phase was considered by a linear relationship between the slow bronchial clearance fraction, f(s), and the geometric particle diameter, derived from bolus inhalation experiments. Potential variations of f(s) from proximal to distal airway generations were simulated by five different scenarios, which allocated slow bronchial clearance to successively peripheral bronchial regions. Alveolar clearance, which contributes only to longterm particle retention, was modeled by transfer rates supplied by the ICRP respiratory tract model. To test the different components of the clearance model, modeling predictions were compared with experimental retention data from bolus inhalation experiments, using various particle sizes and bolus front depths, as well as from slow inhalation experiments, with a flow rate of only 0.045 L sec(-1). The overall good agreement between modeling results and experimental data indicate that the present model correctly predicts bronchial clearance, suggesting that slow bronchial clearance mechanisms are most effective in smaller bronchial airways. PMID- 15120016 TI - Esthetic dentistry in North American dental schools. AB - OBJECTIVES: Esthetic dentistry is among the most dynamic areas of contemporary clinical dentistry. Teaching programs in dental schools have a strong effect on the practice of dentistry, not only for recent graduates, but also for established clinicians, especially with respect to new techniques and concepts. The purpose of the study reported here was to assess the frequency and extent of the teaching of esthetic dentistry in North American dental schools and to report how it differs among the various schools. MATERIALS AND METHODS: A 19-question survey was mailed to 64 North American dental schools. The questions inquired about the priority given to the teaching of esthetic dentistry in the school; how the subject was taught (through regular curricular courses; through a multidisciplinary approach or through elective classes); the duration of the esthetic dentistry course; the nature of the course content (theoretical or practical); the esthetic procedures taught to undergraduate students; the level of interaction among different disciplines in the teaching of esthetic dentistry; and the techniques and commercial materials used. The responses were summarized as percentages based on the number of schools that responded to each question. RESULTS: Fifty-two (81%) of the 64 dental schools completed and returned the questionnaire. Twenty-five of these schools (48%; designated group A) reported having a course exclusively for the teaching of esthetic dentistry. Twenty-seven schools (52%; designated group B) reported that esthetic dentistry was addressed in multiple courses, i.e., no specific course was available. Four schools in group B (15%) were in the process of developing a separate course for esthetic dentistry. In group A schools, esthetic dentistry was taught mainly in the operative dentistry department or division. The most frequent course duration was 4 to 6 months, but there were marked variations. Thirteen (52%) of these 25 schools had didactic and practical teaching at both the preclinical and the clinical levels. The schools in group B reported that only clinical instruction in esthetic dentistry was provided. Several concerns were addressed in the courses offered in group A schools: extrinsic and intrinsic discoloration, bleaching, diastemas, malformation and malpositioning (the latter including rotation, intrusion and labio-linguoversion), and replacement of amalgam and gold restorations. Only 7 (28%) of the group A schools reported having the support of an inhouse laboratory. The esthetic procedures taught were similar for schools in group A and group B. The use of direct posterior composite restorations, all ceramic crowns and nonvital bleaching was more common among group B schools. Ceramic inlays, onlays and indirect posterior composite restorations were not taught by 4 (16%) of the schools in group A and 7 (26%) of the schools in group B. CONCLUSIONS: The teaching of esthetic dentistry in North American dental schools is highly variable and in many schools is shared among different disciplines. Dental schools should work together to establish the parameters for teaching this subject and should formulate the necessary standards for education and research in this new field. PMID- 15120017 TI - University of Alberta dental students' outreach clinical experience: an evaluation of the program. AB - In a 25-year-old satellite dental program, advanced dentistry and dental hygiene students provide highly supervised dental treatment for patients in government funded clinics in 3 under-serviced communities in northern Alberta. Analysis of the performance data of the 2003 University of Alberta dentistry students in their fourth and final year was used to evaluate this program. The data showed that the undergraduate students did numerous diverse general dental procedures during the 2-week rotation, most of which were basic treatments. Students could participate in the program up to 3 times. In subsequent rotations, students tended to do fewer preventive measures, and more restorative treatment and dental extractions. Available grading data for all students indicated that treatment objectives were fulfilled in the vast majority of cases. Such community-based field experience may significantly add to the competence of students entering private practice. PMID- 15120018 TI - Complications of mandibular molar replacement with a single implant: a case report. AB - This case report describes prosthodontic complications resulting from the surgical placement of a single implant and treatment following these complications. Both the surgical and prosthodontic procedures are described for the treatment of a 57-year-old man who had previously received a single implant for the replacement of a missing molar. Using 2 implants, 1 mesial and 1 distal to the previously placed single implant proved reliable. A logical treatment solution is to use 2 implants for the replacement of a single molar to avoid prosthodontic complications. PMID- 15120019 TI - Oral mucosal lesions associated with use of quid. AB - Quid is a mixture of substances that is placed in the mouth or actively chewed over an extended period, thus remaining in contact with the mucosa. It usually contains one or both of 2 basic ingredients, tobacco and areca nut. Betel quid or paan is a mixture of areca nut and slaked lime, to which tobacco can be added, all wrapped in a betel leaf. The specific components of this product vary between communities and individuals. The quid habit has a major social and cultural role in communities throughout the Indian subcontinent, Southeast Asia and locations in the western Pacific. Following migration from these countries to North America, predominantly to inner city areas, the habit has remained prevalent among its practitioners. Many dentists are unaware of the prevalence of the quid or paan habit in the Asian patient population. The recognition of the role of such products in the development of oral precancer and cancer is of great importance to the dental practitioner. A variety of oral mucosal lesions and conditions have been reported in association with quid and tobacco use, and the association of these conditions with the development of oral cancer emphasizes the importance of education to limit the use of quid. In most cases, cessation of the habit produces improvement in mucosal lesions as well as in clinical symptoms. PMID- 15120020 TI - A closer look at diagnosis in clinical dental practice: part 1. Reliability, validity, specificity and sensitivity of diagnostic procedures. AB - Dentists are involved in diagnosing disease in every aspect of their clinical practice. A range of tests, systems, guides and equipment--which can be generally referred to as diagnostic procedures--are available to aid in diagnostic decision making. In this era of evidence-based dentistry, and given the increasing demand for diagnostic accuracy and properly targeted health care, it is important to assess the value of such diagnostic procedures. Doing so allows dentists to weight appropriately the information these procedures supply, to purchase new equipment if it proves more reliable than existing equipment or even to discard a commonly used procedure if it is shown to be unreliable. This article, the first in a 6-part series, defines several concepts used to express the usefulness of diagnostic procedures, including reliability and validity, and describes some of their operating characteristics (statistical measures of performance), in particular, specificity and sensitivity. Subsequent articles in the series will discuss the value of diagnostic procedures used in daily dental practice and will compare today's most innovative procedures with established methods. PMID- 15120021 TI - [Dialog among societies]. PMID- 15120022 TI - [Satellite symposium: Asthma in the World. Asthma in Spain]. PMID- 15120023 TI - Satellite symposium: Asthma in the World. Asthma among children in Europe. PMID- 15120024 TI - [Satellite symposium: Asthma in the World. Asthma among children in Latin America]. AB - The prevalence of respiratory symptoms related to asthma in children from Latin America has been largely ignored. This region participated in phases I and III of the International Study of Asthma and Allergies in Childhood (ISAAC) with 17 participating centers in phase I and 78 centers in phase III. Data were obtained on asthma, rhinitis and eczema from countries and centers with markedly different climactic, cultural and environmental conditions and socioeconomic development. The results for phase I are presented herein because data from phase III are currently being revised at the ISAAC international data control center and will be officially available in the second half of 2004. Phase I provided important information on the prevalence of asthma in the participating countries and demonstrated wide variation among centers in the same country and among countries. The participating Latin American countries are all developing countries and share more or less the same problems related to low socioeconomic status. Therefore, the results and figures should be analyzed within that context. The range for accumulative and current asthma symptoms in children from the Latin American countries that participated in phase I (89,000) were as follows: the prevalence of asthma ranged from 5.5% to 28% in children aged 13-14 years and from 4.1% to 26.9% in children aged 6-7 years. The prevalence of wheezing in the previous 12 months ranged from 6.6% to 27% in children aged 13-14 years and from 8.6% to 32.1% in children aged 6-7 years. The high figures for asthma in a region with a high level of gastrointestinal parasites infestation, a high burden of acute respiratory and gastrointestinal infections occurring early in life, severe environmental and hygiene problems, suggest that these factors, considered as protective in other (developed) regions of the world, do not have the same effect in this region. Furthermore, those aggressive environmental conditions acting together from very early in life might condition different asthmatic phenotypes with more severe clinical presentation in infancy (first 2 years of life), lower atopy and enhanced airways reactivity. The present study indicates that the prevalence of asthma and related symptoms in Latin America is as high and variable as described previously for industrialized or developed regions of the world and that the environmental risk factors, mainly related with poverty, could be responsible for the different clinical and functional presentations of asthma in children from developing regions. PMID- 15120026 TI - [Round table: Importance of allergens in childhood asthma. Introduction]. AB - Asthma is the most frequent chronic disease in childhood. Its prevalence varies (in Spain between 12 % in coastal regions and 6 % in the Castilian plateau). Asthma represents chronic inflammation of the airways associated with an increase in bronchial reactivity to various stimuli, leading to reduced air flow and subsequent remodeling of the bronchial wall. Asthma usually begins in the first few years of life and 30 % of patients will continue to be asthmatic throughout their lives. The main characteristic associated with the persistence of asthma in adulthood seems to be allergy. Another risk factor is the presence of bronchial hyperreactivity. Wheezing associated with viral infections in childhood shows a favorable outcome when not associated with atopic features. The atopic disposition is caused by the interaction between genetic and distinct environmental factors (allergens), as well as by the various clinical manifestations that favor a mainly Th2 response, with interleukins that lead to the formation of specific IgE, pro-inflammatory cytokines, bronchial hyperreactivity, etc. In most cases, asthma is mainly an allergic process, mediated by an IgE mechanism. Inhalation of allergens is considered the most important cause of the onset of asthma in predisposed children. First the presence of an allergen gives rise to sensitization, with the formation of specific IgE. In a subsequent contact, mediators are released, with inflammatory response of the airways and an increase in bronchial hyperreactivity. If the presence of the allergen is prolonged, inflammation and subsequent healing may lead to irreversible damage to the bronchial wall and permanent deterioration of respiratory function. In children with asthma, the aim is to modify the natural history of the disease and prevent its persistence in adulthood. This involves identifying the allergen or allergens causing the symptoms, confirming the role of these allergens in clinical manifestations through specific challenge tests and, lastly, indicating the treatment be followed. PMID- 15120025 TI - [SENP-SEICAP (Spanish Society of Pediatric Pneunomology. Spanish Society of Pediatric Clinical Immunology and Allergology) consensus on asthma, pneumonology, and pediatric allergy (Draft)]. PMID- 15120027 TI - [Allergenic pollens in Spain]. AB - Allergenic pollens that cause rhinoconjuctivitis and/or asthma are those from trees or plants that pollinate through the air (anemophilic pollination) and not through insects (entomophilic pollination). Although pollen grains would seem to be too large to easily reach the intrapulmonary airways, the relationship between pollen counts and the presence of asthmatic symptoms is only too evident. This is probably because the allergens inducing seasonal asthma are not only found within pollen grains but also outside the grains in particles of less than 10 mm that are freely found in the atmosphere. The most important pollens producing pollinosis in Spain are those from cypress trees from January-March, birch trees in April (macizo galaico), Platanus hispanica (March-April), grasses and olive trees from April-June, Parietaria from April-July and Chenopodium and/or Salsola from July-September. By geographical areas, the main cause of pollinosis are grasses in the center and north of the peninsula, olive trees in the south (Jaen, Sevilla, Granada, Cordoba) and Parietaria in the Mediterranean coast (Barcelona, Murcia, Valencia). PMID- 15120028 TI - [Clinical importance of cross-reactivity between allergens]. AB - Cross reactivity describes the development of symptoms as a consequence of the presence of specific IgE to a protein without the subject having had previous contact with that protein. Cross-reactivity depends on factors such as the individual's immune response, the type and intensity of exposure and, above all, the type of allergen. The identification of pan-allergens, which are present in various animal and vegetable sources and which show great structural and sequential similarity, even among species with little taxonomic relation, explains the existence of distinct, well-defined cross-reactivity syndromes. Knowledge of these phenomena could have important diagnostic and therapeutic consequences. PMID- 15120029 TI - [Indications and utility of allergen provocation testing]. AB - Specific bronchial provocation tests have been used to confirm diagnosis, as a laboratory model for the study of asthma, and to evaluate the efficacy of immunotherapy with allergen extracts. However, despite the advances achieved in our knowledge of the etiopathogenic mechanisms of asthma, bronchial provocation tests with allergens have certain methodological limitations. These are due to the difficulty of evaluating these tests as regards which markers of inflammation (exhaled nitric oxide, carbon monoxide, etc.) should be investigated and at what points in the chronology of the asthmatic response. The current parameters used to assess respiratory function (FEV1, mainly) do not seem to be the most ideal nor do their percentage reductions, usually established by use, always serve to identify a bronchospastic and inflammatory response. Furthermore, the allergen doses indicated in the various protocols do not mimic natural exposure to these allergens, thus introducing an important bias in the interpretation of the results. In this sense, another and no less important difficulty is the lack of adequate standardization of the characteristics of the allergens used; the development of recombinant allergens may eliminate this obstacle and improve the reproducibility of the tests. In view of the above, the requirement of a positive bronchial provocation test to a specific allergen in the etiological diagnosis of asthma does not seem reasonable nor does it seem reasonable that specific immunotherapy is not indicated if the test is negative or that evaluation of its efficacy lies in a negative result for a previously positive test. This premise does not take into account the fact that bronchial reactivity is subject to variables that are difficult to control (prior natural exposure to the allergen, for example). PMID- 15120030 TI - [Efficacy of immunotherapy in the treatment of asthma]. AB - Specific immunotherapy consists of the administration of allergen extracts to patients with allergic disease to achieve clinical tolerance to the causative allergens. Currently, it is the only etiologic treatment for respiratory allergy. A World Health Organization opinion paper published in 1997 defines immunotherapy as "the only form of treatment able to modify the natural course of allergic diseases". In patients with allergic rhinitis, several studies suggest that immunotherapy can modify the natural history of respiratory allergy by preventing the development of asthma in children with this disease. Numerous studies demonstrate its efficacy in IgE-mediated asthma and particularly in mild-to moderate asthma. When complete avoidance of the allergen cannot be achieved with measures that allow the patient to lead a normal life, pharmacological treatment can help to control symptoms, but symptoms immediately return when treatment is interrupted. However, asthma care can be improved by allergen-specific treatment; immunotherapy may shift the immune response from an allergic pattern toward a more protective response, producing persistent improvement with reduction of symptoms and the need for pharmacological treatment. Numerous comparative studies with specific immunotherapy vs. placebo or pharmacological treatment have demonstrated the efficacy of this treatment and its advantages in control of the disease. Specific immunotherapy induces favorable clinical, biological and functional modifications in the course of allergic asthma. Significant improvement in clinical manifestations has been demonstrated, even with levels of allergen exposure higher than those at the beginning of treatment. This improvement is associated with a reduced need for antiinflammatory and bronchodilator treatment. Moreover, specific bronchial reactivity shows a clear improvement with disappearance of delayed response and a clear increase in the threshold for immediate response to the allergen. Reduction in nonspecific bronchial hyperreactivity and improvement in exercise-induced asthma are also observed. Several studies recommend an optimal duration of specific immunotherapy for allergic asthma of between 3 and 5 years to achieve maximal therapeutic efficacy. A direct relationship between treatment duration and the persistence of its effects has been observed. Moreover, the treatment is more effective when started early. The possible adverse effects related to systemic reactions should be borne in mind. Although these effects are infrequent, maximal precautions should be taken when administering this treatment. Immunotherapy is contraindicated in cases of severe asthma, heart disease, autoimmune disease and associated severe neoplastic processes. However, all the beneficial effects of immunotherapy are conditioned by an accurate and early etiological diagnosis confirming the causative allergen. The availability of high-quality allergen extracts is essential to obtain the desired effect. Inappropriate patient selection for this treatment is the main cause of its failure. The integral treatment of allergic asthma includes environmental measures, patient education, pharmacological treatment and, whenever possible, immunotherapy. PMID- 15120034 TI - Update on NCI in vitro drug screen utilities. AB - Development of new anti-cancer drugs is a costly and risky proposition. The Developmental Therapeutics Program (DTP) of the National Cancer Institutes of the United States (U.S.) facilitates the drug development process by providing access to preclinical screening services. Since the early 1990's, DTP has screened tens of thousands of compounds against a panel of 60 human tumour cell lines representing nine tissue sites. At the same time, DTP began to accumulate information on the expression of molecular entities in the same 60 cell line panel. Many of these data are freely available to the public at. More recently, additional, more focused screens have entered the picture, with data also available through the web site. These include screening of roughly 100000 compounds against a panel of yeast mutants, and screening of the NCI Diversity Set in assays designed to detect effects on Molecular Targets of interest. PMID- 15120035 TI - In vitro modelling of human tumour behaviour in drug discovery programmes. AB - Human tumour cell lines have played an important part in our understanding of cancer and have been used extensively in the discovery and characterisation of new chemotherapeutic drugs. A potential weakness of such cell lines is that they may have lost important properties originally possessed in vivo, including potential targets for therapy. This review discusses how possible differences between tumour cells in cancer patients and cell lines might be identified by the use of short-term cultures of human tumour cells taken directly from cancer tissue, termed here primary cultures. Cell-cycle time is one important difference between tumours and cell lines and it is known that the cell-cycle times of primary cultures cover the same wide range as estimated in vivo cell-cycle times. Because tumour cells have at least two pathways to cell death, one from interphase and one from mitosis, changes in cell-cycle length can modify the balance of such pathways. Responses of primary cultures to DNA-damaging drugs and inhibitors of growth factor receptors also differ from those of cell lines, suggesting that the process of developing a cell line can result in the loss of important cellular responses. Without an appreciation of these changes our ability to discover new targets for the development of improved cancer therapy may be jeopardised. The identification of cell lines that preserve potential targets is an important goal in cancer biology and research using primary cultures will help in this identification. PMID- 15120036 TI - Clonogenic assay with established human tumour xenografts: correlation of in vitro to in vivo activity as a basis for anticancer drug discovery. AB - Pluripotent cells can be grown in clonogenic assays. The tumour stem-cell fraction, which accounts for <0.4% of the total cells, and which is considered the most relevant cell type in the development of metastases and recurrences, is able to divide and to form colonies in a semisolid matrix (agar or methylcellulose). Major applications of the tumour clonogenic assay (TCA) are chemosensitivity testing of tumours and xenografts, and for assessments within drug discovery programmes. Of critical relevance for the usefulness of the TCA is whether it can predict sensitivity or resistance towards clinically used agents. When we compared the response of human tumours established as xenografts in nude mice in the TCA in vitro to that of the clinical response, 62% of the comparisons for drug sensitivity, and 92% of the comparisons for drug resistance were correct. The same percentage of true/false observations was found when tumours were tested after serial passage in nude mice in the TCA in vitro and their response compared to in vivo activity in corresponding xenografts (60% and 90%, respectively). The highest correct predictive values were, however, found when the clinical response of tumours was compared to their explants established in the nude mouse and treated in vivo. Of 80 comparisons performed, we observed a correct prediction for tumour resistance in 97% and for tumour sensitivity in 90%. In our opinion, the TCA with established human tumour xenografts has an important role in current drug discovery strategies. We therefore included the TCA as secondary assay in our approach to anticancer drug discovery and found that a number of novel agents were active; these are now in advanced preclinical development or clinical trials. Thus, the tumour clonogenic assay has proven predictive value in the chemosensitivity testing of standard and experimental anticancer drugs. PMID- 15120037 TI - The hollow fibre model in cancer drug screening: the NCI experience. AB - The in vivo hollow fibre model was developed by the National Cancer Institute (NCI) in the United States of America (USA) at a time when the number of potential anti-cancer drugs arising from in vitro screening efforts exceeded the available capacity for testing in traditional xenograft models. Updated analysis of the predictive value of the hollow fibre model continues to indicate that the greater the response in the hollow fibre assay, the more likely it is that activity will be seen in subsequent xenograft models. The original 12 cell line hollow fibre panel has been supplemented with histology-specific panels, and we begin here to analyse their utility in predicting activity in subsequent in vivo models. The key goal of using the hollow fibre model as a way to decrease the cost, both financial and in the number of animals used, to evaluate initial evidence of a compound's capacity to act across physiological barriers continues to be reinforced with our enlarging experience. PMID- 15120038 TI - Of mice and men: values and liabilities of the athymic nude mouse model in anticancer drug development. AB - Human tumour xenografts implanted subcutaneously (s.c.) into immunosuppressed mice have played a significant role in preclinical anticancer drug development for the past 25 years. Their use as a predictive indicator of probable clinical activity has been validated for cytotoxics. A retrospective analysis for 39 compounds where both extensive xenograft testing and Phase II clinical data were available, performed by the National Cancer Institute (NCI), has shown that 15/33 agents (45%) with activity in more than one-third of xenografts showed clinical activity (P=0.04). However, with the exception of non-small cell lung cancer, activity within a particular histological type of the xenograft generally did not predict for clinical activity in the same tumour. Today, the question (largely unanswered) is how useful is the xenograft model (particularly the traditional s.c. model) in contemporary cancer drug discovery? There are many variables when conducting xenograft experiments which impact on outcome; viz, site of implantation, growth properties of the xenograft and size when treatment is initiated, agent formulation, scheduling, route of administration and dose and the selected endpoint for assessing activity. The xenograft model remains of value in current preclinical cancer drug development, especially when such studies give due consideration to the above variables and are based on sound mechanistic (e.g. status of the selected target in the chosen model) and pharmacological (e.g. use of formulated agent) principles. Dependent upon the drug target, a slowing of xenograft tumour growth (cytostatic effect) rather than tumour shrinkage might be the major observed effect. Human tumour xenografts are also particularly useful in determining pharmacodynamic markers of response for subsequent clinical application. Nevertheless, it needs to be kept in mind that the use of xenografts is relatively time-consuming and expensive, raises animal ethical issues and there are instances where the model is inappropriate as a likely predictor of clinical outcome (e.g. inhibitors of the metastatic process and anti-angiogenic strategies as the vasculature is of murine origin). PMID- 15120039 TI - Integrating pharmacology and in vivo cancer models in preclinical and clinical drug development. AB - Historically, cancer drug development has been a roller coaster. Numerous agents have shown exciting activity in preclinical models and yet have had minimal activity clinically. These disappointments have led to reasonable scepticism about the true value of both syngeneic and xenograft rodent tumour models in accurately identifying agents that will have important clinical utility. Whereas the development of newer techniques, including transgenic mouse models of cancer, offers the potential to develop more predictive models, the role of such mice in cancer drug development is not yet validated. To advance in our understanding of predictive model systems it may be wise to analyse both the successes and the failures of conventional models in order to understand some of their limitations and perhaps to avoid making the same mistakes in the future. Here we review the value and limitations of xenograft models, and the role of integrating preclinical pharmacology in developing new treatments for solid tumours of childhood. PMID- 15120040 TI - In vivo models for endocrine-dependent breast carcinomas: special considerations of clinical relevance. AB - Tumours in hormone-regulated organs such as the breast, prostate or ovaries are among the most frequent malignancies. Because of their endocrine-dependent development and growth, they offer a unique opportunity for antihormonal treatment either single or long-term or in combination with radio- or chemotherapy. A prominent example is breast carcinoma, for which the anti oestrogen tamoxifen has been used successfully for several years. Unfortunately, a substantial number of tumours are intrinsically tamoxifen-resistant, despite oestrogen-receptor positivity, and, eventually, almost all breast carcinomas acquire resistance towards tamoxifen. The recently developed pure anti-oestrogen Faslodex and the third-generation aromatase inhibitors (Letrozol, anastrozole (Arimidex) offer the possibility of alternative therapies. Preclinical models are needed, as most of the mechanisms of hormonal tumour dependence and the causes of the appearance of antihormone resistance are not yet fully understood. This review focuses on the development and characterisation of breast cancer xenografts derived directly from surgical resections. With their help, a deeper insight into the mechanisms of hormone regulation and anti-oestrogen resistance can be gained. The xenograft models have already been used in differential gene expression analysis on DNA microarrays and for the evaluation of approaches to overcoming tamoxifen resistance. PMID- 15120041 TI - Orthotopic models of cancer for preclinical drug evaluation: advantages and disadvantages. AB - Considering the enormous effort that has taken place over the years to discover new chemotherapeutic drugs for treating the common cancers, the conventional murine and xenograft test systems used to test efficacy for drug development have identified only a limited number of useful agents that are active clinically at well tolerated doses. In recent years, considerable effort has been made to develop more clinically relevant models by the use of orthotopic transplantation of tumour material in rodents. It has been shown that it is now possible to transplant tumour material from a variety of tumour types into the appropriate anatomical site and often these tumours will metastasise in a similar manner and to similar locations as the same tumour type will in human cancer. As yet, although a body of literature has amassed on the technique itself and its implications for metastasis, there are relatively few laboratories using these test systems in drug development programmes. Nevertheless, given the expertise now being developed and some interesting observations being made on the role of the tumour site on response to therapeutic agents, it is likely that the use of orthotopic systems will strengthen our ability to select the most appropriate molecules for recommended use in clinical studies. PMID- 15120042 TI - Spontaneous and genetically engineered animal models; use in preclinical cancer drug development. AB - The preclinical development of anticancer drugs has been based primarily on the transplantation of murine or human cancers into mice. Alternatives to these transplantation models are animals that naturally develop cancers with features relevant to the human disease. The first group of these models arises in mice that are genetically engineered to develop cancer. The second group includes pet dogs and cats that naturally develop cancer. This review will discuss the use and integration of these spontaneous cancer models into a comprehensive and comparative approach to preclinical drug development. Examples of their successful use and an outline of their relative strengths and weaknesses will be provided. PMID- 15120043 TI - Modelling approaches for angiogenesis. AB - The development of a functional vasculature within a tumour is a requisite for its growth and progression. This fact has led to the design of therapies directed toward the tumour vasculature, aiming either to prevent the formation of new vessels (anti-angiogenic) or to damage existing vessels (vascular targeting). The development of agents with different mechanisms of action requires powerful preclinical models for the analysis and optimization of these therapies. This review concerns 'classical' assays of angiogenesis in vitro and in vivo, recent approaches to target identification (analysis of gene and protein expression), and the study of morphological and functional changes in the vasculature in vivo (imaging techniques). It mainly describes assays designed for anti-angiogenic compounds, indicating, where possible, their application to the study of vascular targeting agents. PMID- 15120044 TI - A potential role for imaging technology in anticancer efficacy evaluations. AB - The introduction of imaging methods suitable for rodents offers opportunities for new anticancer efficacy models. Traditional models do not provide the level of sensitivity afforded by these precise and quantitative techniques. Bioluminescent endpoints, now feasible because of sensitive charge-coupled device cameras, can be non-invasively detected in live animals. Currently, the most common luminescence endpoint is firefly luciferase, which, in the presence of O(2) and ATP, catalyses the cleavage of the substrate luciferin and results in the emission of a photon of light. In vivo implantation of tumour cells transfected with the luciferase gene allows sequential monitoring of tumour growth within the viscera by measuring these photon signals. Furthermore, tumour cell lines containing the luciferase gene transcribed from an inducible promoter offer opportunities to study molecular-target modulation without the need for ex vivo evaluations of serial tumour samples. In conjunction with this, transgenic mice bearing a luciferase reporter mechanism can be used to monitor the tumour microenvironment as well as to signal when transforming events occur. This technology has the potential to reshape the efficacy evaluations and drug-testing algorithms of the future. PMID- 15120045 TI - The Cancer Research UK experience of pre-clinical toxicology studies to support early clinical trials with novel cancer therapies. AB - Pre-clinical toxicology studies in rodents and Phase I clinical trial data are summarised for 14 novel anticancer therapies. With only one exception, an antifolate antimetabolite, rodent toxicology predicted a safe Phase I trial starting dose and the majority of the dose limiting toxicities, in particular haematological toxicity. For targeted agents with well-defined pharmacodynamic markers, illustrated in the current study by 3 anti-endocrine drugs and one resistance modifier, the definition of a maximum tolerated dose can be avoided. Together with earlier data, the current study confirms that pre-clinical toxicology studies in a non-rodent species are not routinely needed for the safe conduct of early clinical trials with new cancer chemotherapies. PMID- 15120046 TI - Multi-species toxicology approaches for oncology drugs: the US perspective. AB - The Toxicology and Pharmacology Branch (T&PB) of the National Cancer Institute (NCI) performs pharmacological and toxicological evaluations of new oncology agents according to an agent-directed paradigm in which all studies are tailored to each agent. The United States Food and Drug Administration (US FDA) requires that preclinical toxicology studies be conducted in two species, a rodent and a non-rodent for all small molecules, and T&PB has successfully used this formula. While pharmacokinetic (PK) studies are considered optional, T&PB routinely develops new methods for plasma/tissue drug analysis and employs this methodology throughout development to determine kinetics in various species and toxicokinetics in the toxicity studies. In the current era of molecular target based development, the T&PB also develops or employs methodology to evaluate effects of the new chemical entity on appropriate biomarkers in tumour and normal tissues. In this comprehensive programme, T&PB is able to correlate safety and toxicity with both plasma drug levels and biomarker modulation in two species for a seamless entry into Phase I. PMID- 15120047 TI - Do we understand who benefits from resynchronisation therapy? PMID- 15120048 TI - Beta-blockers, myocardial ischaemia and collateral circulation. PMID- 15120049 TI - Reflections on the Danish Revolution. PMID- 15120050 TI - Prognostic relevance of symptoms versus objective evidence of coronary artery disease in diabetic patients. AB - AIM: Little is known about the prognostic significance of silent versus symptomatic coronary artery disease (CAD) in diabetic patients. We therefore assessed the incidence of scintigraphic evidence of CAD in diabetic patients without known CAD and the impact of symptoms and scintigraphic findings on prognosis. METHODS AND RESULTS: A consecutive series of 1737 diabetic patients without known CAD underwent dual-isotope myocardial perfusion SPECT (MPS) and 1430 were followed-up for a median of 2 (1-8.5) years. Critical events were defined as myocardial infarction or cardiac death. Objective evidence of CAD was found in 39% of 826 asymptomatic diabetic patients, in 51% of 151 diabetic patients with shortness of breath (SOB), and in 44% of 760 diabetic patients with angina. During follow-up, 98 critical events occurred. Annual critical event rates were 2.2% in asymptomatic, 3.2% in angina, and 7.7% in diabetic patients with shortness of breath ( p < 0.001 versus other groups). With MPS evidence of CAD, critical event rates increased to 3.4% (asymptomatic), 5.6% (angina), and 13.2% (SOB) ( p 50 years and controls (p = 0.0002 and p = 0.0001). We estimated and compared the haplotype frequencies between C270T and G196A markers in PD and controls that was positive (p = 0.0019). All positive results remain significant after Bonferroni's correction. Our data indicate the possibility of linkage disequilibrium between the C270T variation and a mutation in coding region of the BDNF gene and suggest that this gene may play a role in the development of familial PD. PMID- 15120096 TI - Human brain dopamine metabolism in levodopa-induced dyskinesia and wearing-off. AB - The objective of this study was to identify dopamine (DA) metabolism pattern in Lewy body Parkinson's disease (PD) patients with dyskinesia (Dysk) only, with wearing-off (WO) only, or no motor complications (NMC) induced by levodopa (LD). DA, homovanillic acid (HVA), 3,4-dihydroxyphenylacetic acid (DOPAC), and 3 methoxytyramine (3-MT) were measured individual basal ganglia nuclei of nine PD patients who received LD for 6-18 years. Three patients had only Dysk, three only WO, and three had neither Dysk nor WO. Biochemical measurements in PD brains were compared with four non-neurological control brains from individuals matched for age and post-mortem retrieval time. DA levels in the PD were reduced in the caudate by 87% and putamen by 99%. In the caudates, the HVA/DA molar ratio as an index of DA metabolism was similar in the WO and the Dysk patients. However, in the putamen, the ratio of HVA/DA was significantly higher in the WO compared with the Dysk (p = 0.03)and the NMC (p = 0.04) groups of patients. In the putamen, the DOPAC levels were higher in the WO cases while in the Dysk cases, 3-MT levels were higher. The results suggest that in the WO only cases, the putaminal DA was in large measure metabolized intraneuronally while the DA metabolism in our Dysk only patients was mainly extraneuronal. We conclude that the magnitude and the site (intra vs. extraneuronal) of the synaptic DA metabolism in the putamen plays a significant role in LD-induced Dysk and WO. PMID- 15120097 TI - Visual object recognition deficits in early Parkinson's disease. AB - The nature of the visual perception deficits in Parkinson's disease (PD) has remained unclear. The present study explored whether there emerge deficits in the different stages of visual object recognition in early PD. Twenty-eight patients and 14 healthy controls were studied. A set of reaction time tasks were applied to measure the different stages of object recognition. The results indicate some selective problems in both basic perceptual and semantic visual processing at an early stage of cognitive deterioration in PD. PMID- 15120099 TI - Flunarizine and cinnarizine-induced parkinsonism: a historical and clinical analysis. AB - BACKGROUND: Drug-Induced Parkinsonism (DIP) represents the second leading cause of Parkinsonism (PK) in several countries. Flunarizine and cinnarizine are some of the most common drugs that cause DIP. This paper reviews the first description of Flunarizine and Cinnarizine-Induced Parkinsonism (FCIP), as well as the subsequent literature, emphasizing epidemiological, clinical and diagnostic aspects. METHODS: We reviewed the literature on the subject, with special emphasis on the first description and the later definition of the clinical syndrome that results from chronic use of flunarizine and cinnarizine. RESULTS: In 1984, De Melo-Souza reported the first description of flunarizine-induced PK in five patients. Other reports followed on FCIP, emphasizing the clinical features, which are symmetrical parkinsonism, and depression, affecting mainly elderly women. CONCLUSIONS: Eighteen years after the original description, FCIP is a recognized condition with specific clinical features, and is the second most common cause of parkinsonism in many countries. PMID- 15120098 TI - Therapeutic factors causing hallucination in Parkinson's disease patients, especially those given selegiline. AB - Selegiline protects nigral dopaminergic neurons and is recommended for the treatment of patients in the early stage of Parkinson's disease (PD). We treated 112 PD patients and noted that those given selegiline had a high incidence of hallucination. Our objective was to determine which clinical therapeutic factors cause such hallucinations. The Kruskal-Wallis and chi-square test showed that in 94 patients, the severity of the hallucinations was significantly related to the duration of illness, Hoehn and Yahr stage, doses of levodopa and cabergoline, whether or not selegiline was used, and whether or not medication for constipation was required. In addition, patients who were treated with a low dose of levodopa (< or =300 mg/day), who had a low Hoehn and Yahr stage, and a short duration of illness (< or =8 years) together with a high dose of selegiline or cabergoline also tended to have hallucinations. MRI findings were not related to the incidence of hallucination. When selegiline is given to patients who have PD of long duration and a high Hoehn and Yahr stage, and who already are receiving levodopa and a dopamine agonist, the doses of levodopa and the dopamine agonists given, as well as the presence of constipation, may be related to the incidence of hallucination. PMID- 15120100 TI - Explosive-aggressive behavior related to bilateral subthalamic stimulation. AB - We describe a patient with Parkinson's Disease who underwent bilateral subthalamic nucleus deep brain stimulation and later presented with episodes of aggressive behavior disorder with disturbed impulse control and an inability to control anger likely related to the deep brain stimulation "switch-on stimulation". We hypothesize that increasing voltage intensity could influence neighboring passing fibers coming from basal limbic system that are involved in the regulation of affect and emotional behavior. We suggest investigating these neuropsychological disturbances considering their influence on quality of life after surgery. PMID- 15120101 TI - Olfactory and visual hallucinations in Parkinson's disease. AB - Management of hallucinations in patients with Parkinson disease is a challenge for both the physician and the patient. They occur in more than 30% of patients and they present mostly as visual hallucinations but other forms such as auditory and tactile hallucinations have been reported. We have reported an interesting patient with olfactory hallucinations and visual hallucinations. To our knowledge, olfactory hallucinations have been rarely described in patients with Parkinson's disease and might be added to the late complications of Parkinson's disease. PMID- 15120102 TI - Head-turning dizziness in multiple system atrophy. PMID- 15120103 TI - [Hepatitis G virus and labile blood products: role of transfusional transmission]. AB - The GBV-C/HGV (HGV) virus was discovered a few years ago. This virus is known to be parenterally as well as sexually transmitted. However, no study has found some pathogenic roles for HGV so far. In the present study, we aimed to investigate the transmission of HGV by blood components transfused to 284 patients hospitalized in surgery unit in 1995. We tested two parameters of infection in blood components transfused to infected recipients: viral RNA by PCR and anti-E2 antibodies by ELISA. We tried to suspect some potent hepatocyte impacts by assessing the levels of two enzymes in serums: alanine aminotransferase (ALT) and alpha-glutathion S-transferase (alpha-GST). We found that HGV-RNA was detectable in 3.6% of recipients prior to transfusion and 7.5% post-transfusion. For each infected recipient, we retrospectively did a search for HGV-RNA in each transfused blood component, and we found at least one blood component as HGV-RNA positive for each transfusional infected recipient. Anti-E2 antibody prevalence standing for a former and cured infection was 39.6% in all the recipients. In viremic recipients, ALT levels were mostly normal, while alpha-GST levels were found more commonly elevated than in non-viremic recipients although non significantly (20% vs. 6.3%; P = 0.07). The present study underlines that HGV transmission is mostly transfusional in surgery units, and that infectiosity of blood components can be anticipated by detection of the viral RNA by PCR. Furthermore, the possible relationship between the serum activity of alpha-GST and the hepatotropism of HGV, although non-admitted as pathogenic, should be investigated. PMID- 15120104 TI - [Trends in residual risk of transfusion-transmitted viral infections (HIV, HCV, HBV) in France between 1992 and 2002 and impact of viral genome screening (Nucleic Acid Testing)]. AB - BACKGROUND: Monitoring trends in residual risk of transfusion-transmitted viral infections is important to assess improvements in blood safety. These trends were analysed over nine overlapping periods of 3 years from 1992 to 2002. The 2000 2002 estimates were compared to the results of HIV-1 and HCV NAT implemented on all blood donations in July 2001. METHOD: As risk is mainly associated with the window period, residual risks were estimated by multiplying incidence rates by the durations of the window periods. For the first seven periods, incidence rates were calculated from data collected by the blood centres belonging to the Transfusion-Transmissible Agents Working Group which collect more than 50% of blood donations in France, and for the two last periods, on the overall blood supply. RESULTS: On the 2000-2002 period, residual risks without NAT were estimated at 1 in 1,400,000 for HIV, at 1 in 1,000,000 for HCV and at 1 in 400,000 for HBV. With minipool NAT, the residual risk become nearly two times lower for HIV (1 in 2.5 million donations) and seven times lower for HCV (1 in 6.65 million donations). For HIV, of the 4.9 million donations screened with NAT between July 2001 and June 2003, two were remote thanks to the NAT, which is consistent with the NAT expected yield. Concerning HCV, one out of the four WP predicted cases was detected with NAT. Without NAT, the overall residual risk for the three viruses combined (HIV, HCV, HBV) decreased from 1 in 65,000 to 1 in 235,000 donations between 1992 and 2002. Since the implementation of NAT, the current overall residual risk is 1 in 325,000 donations (28% less than without NAT). CONCLUSION: NAT results confirm the validity of residual risk estimates given by the model, and the limited benefit of genomic screening due to the very low level of residual risk at the time of its implementation. PMID- 15120105 TI - [Post-transfusion malaria: is the risk irreconciliable with biological silence?]. AB - Despite the relatively high frequency of imported malaria in metropolitan France, the transmission of malaria by transfusion is exceptional. The screening of donations to determine those at risk is performed by an interview, and by the testing of serology for defined groups of donors. However, the exclusion of a candidate 'at risk' as a blood donor, by a pre-donation interview, is not completely mastered and the discrimination by biological examination lacks sensitivity, as much for methodological reasons as for reasons linked to the complex parasitic pathogenic agent (Plasmodium ssp.), as for the specific host defence system. The risk of introducing an unsafe-potentially dangerous (transfusion-transmitted malaria is often lethal)-element into the transfusional circuit is not completely covered. Is serology testing the most adequate test to avoid the risk of infected donations, in particular by Plasmodium falciparum; what are the alternatives and what will be the eventual added-costs of the biological qualification of such donations? The transfusional risk linked to Plasmodium seems, however, to be reduced to a minimum, concerning the circulation of plasma, which could represent an alternative for donors at real risk (rare) and those with a supposed risk (relatively numerous). PMID- 15120106 TI - [ABO polymorphism in blood donors in Morocco]. AB - Blood grouping were improved since the advent of monoclonal antibodies and the automation of the tests. No comparative study has been done before in Morocco concerning ABO regional genetic frequencies. Hence, the aim of this work is to actualize and to precise them. The ABO blood grouping has been carried out on 344 954 blood donors by micro method in several regional blood transfusion centers. Genetic frequencies were calculated according to the Bernstein's method. The national mean values are A: 0.2141, B: 0.105, O: 0.6777. The regional frequencies show gradients between the north and the south of Morocco. PMID- 15120107 TI - [Evaluation of residual infectious risk among blood donors in National Center of Blood Transfusion in Conakry]. AB - To estimate the risk of transmitting human immunodeficiency virus (HIV) and hepatitis B virus (HBV) by blood transfusion. Residual risks for each of infections have been calculated from incidence cases number, rate incidence for 100,000 person-years has been estimated and multiplied by the period of mute serological window for each agent (22 days for HIV and 56 for HBV) in order to estimate the residual risk. This study shows that risk residual is 1/121 blood donations for HBV and 1/8562 blood donations for HIV. The obtained results show that the HIV and HBV transmission risk due to blood transfusion according to the present practice at the NCBT of Conakry is important. PMID- 15120108 TI - [Legal obligation to inform the patient on the theoretical risk of CJD transmission by blood]. AB - Legal obligation to inform the patient does not include theoretical risks. However, due to the very sensitive situation of blood transfusion in France, following the tainted-blood affair, a circular was issued to extend this obligation (1998) to inform the theoretical risk of CJD transmission by blood. Ethically speaking, this raises three questions: Is it beneficial to the patient to be informed on theoretical risk? Is the use of a "circular", less legally binding, appropriate? Finally, what is the situation in other countries? The evolution of the law tends to be more positive in that it no longer involves any theoretical risk. PMID- 15120109 TI - [Current data on granulocytes donations]. AB - Granulocyte transfusion is uncommon but essential for some aplastic patients with major infection and those with septic granulomatosis disease and visceral aspergillosis. Compatibility between donor and recipient (ABO, D, Kell blood type, cytomegalovirus) is necessary. Stimulation by dexamethasone, before use of GCSF, permits to obtain approximately 10 x 10(9) white cells per l. Doses of heparin must be calculated to ensure an efficient hypocoagulation without any trouble for the donor immediately after his donation. Sedimentation by hydroxyethylstarch is an unavoidable element to obtain the most efficient separation of granulocytes. Most donations present an amount of white cells between 2 and 4 10 x 10(10). This enables to transfuse efficiently children and adults. PMID- 15120110 TI - The thioredoxin h system of higher plants. AB - In plants, thioredoxins h are encoded by a multigenic family of genes (eight in Arabidopsis thaliana, at least five in Populus sp.). The multiplicity of these isoforms raises the question of their specificity. This review focuses on thioredoxins h in two plant models: Arabidopsis and poplar. Thioredoxins h can be divided into three different subgroups according to the analysis of their primary structure. This paper describes the biochemical properties of each subgroup. Recent data in the field indicate that subgroup members differ by their subcellular localization as well as their reduction pathways suggesting specific functions for each subgroup. The development of proteomic tools has also increased considerably the number of potential thioredoxin targets, showing the importance of thioredoxins h in plants. PMID- 15120111 TI - Sensing and signalling during plant flooding. AB - Flooding is a major issue for plant survival in many regions of the world. Soil inundation induces multiple plant physiological dysfunctions, leading to a decline in plant growth and survival capacity. Some of the most important effects of flooding include a reduction in water and nutrient uptake and a decrease in metabolism. Prolonged soil flooding will also ultimately lead to anoxia conditions with profound effects on plant respiratory metabolism. However, it is still unclear which signals and which sensory mechanisms are responsible for triggering the plant response. In contrast, it is now established that flooding responses are typified by enhanced ethylene production, accompanied by a signalling cascade which includes a network of hormones and other common secondary signalling molecules. In recent years, there has been significant progress in the understanding of some of the signalling pathways involved during plant stress responses. Here, we present an overview of recent hypothesises on sensing and signalling during plant flooding. PMID- 15120112 TI - The plant uncoupling protein homologues: a new family of energy-dissipating proteins in plant mitochondria. AB - Uncoupling proteins (UCPs) form a subfamily within the mitochondrial carrier protein family, which catalyze a free fatty acid-mediated proton recycling and can modulate the tightness of coupling between mitochondrial respiration and ATP synthesis. As in mammalian tissues, UCPs are rather ubiquitous in the plant kingdom and widespread in plant tissues in which they could have various physiological roles, such as heat production or protection against free oxygen radicals. The simultaneous occurrence in plant mitochondria of two putative energy-dissipating systems, namely UCP which dissipates the proton motive force, and alternative oxidase (AOX) which dissipates the redox potential, raises the question of their functional interactions. PMID- 15120113 TI - Cloning and over-expression of a cDNA encoding a polyketide synthase from Cannabis sativa. AB - A polyketide synthase has been suggested to play an important role in cannabinoid biosynthesis in Cannabis sativa L. This enzyme catalyzes the biosynthesis of olivetolic acid, one of the precursors for cannabinoid biosynthesis. Using a reverse transcriptase-polymerase chain reaction (RT-PCR) based on the DNA homology of chalcone synthase (EC 2.3.1.156) and valerophenone synthase (EC 2.3.1.156) of hop (Humulus lupulus), a cDNA encoding a polyketide synthase in C. sativa was identified. The coding region of the gene is 1170 bp long encoding a 389 amino acid protein of a predicted 42.7 kDa molecular mass and with a pI of 6.04. The gene shares a high homology with a chalcone synthase gene of H. lupulus, 85% and 94% homology on the level of DNA and protein, respectively. Over expression of the construct in Escherichia coli M15 resulted in a 45 kDa protein. The protein has chalcone synthase activity as well as valerophenone synthase activity, a chalcone synthase-like activity. Using n-hexanoyl-CoA and malonyl-CoA as substrates did not give olivetol or olivetolic acid as a product. PMID- 15120114 TI - Sucrose synthase isoforms in cultured tobacco cells. AB - The plant enzyme sucrose synthase (SuSy; EC 2.4.1.13) catalyzes the reversible conversion of sucrose and UDP into UDP-glucose (UDP-Glc) and fructose. The enzyme exists in different isoforms and is both located in the cytosol, membrane-bound and associated to the actin cytoskeleton. We here investigate sucrose synthase from tobacco (Nicotiana tabacum L.) BY-2 heterotrophic cell suspensions. Two different isoforms of sucrose synthase SuSy1 and SuSy2, could be purified from cytosolic extracts of these cells using a combination of poly(ethylene glycol) (PEG) precipitation, gel filtration, ion-exchange chromatography and affinity chromatography. They were clearly distinct, both with regard to the binding to the ion-exchange column and with regard to their kinetic and regulatory properties. SuSy1, the more abundant species, showed lower V(max) and K(m) for sucrose and UDP compared to the less abundant SuSy2. The activity of SuSy2 in the breakdown direction was stimulated by 60% by actin, in contrast to that of SuSy1, which showed a 17% inhibition. An indication of interaction between SuSy1 and actin was obtained by partitioning in aqueous Dextran-PEG two-phase systems. Furthermore, fructose 2,6-bisphosphate (F26BP) at micromolar concentrations stimulated SuSy2 in the presence of actin while SuSy1 was strongly inhibited by fructose. Possible roles of these two isoforms in the sucrose turnover in BY-2 cells are discussed. PMID- 15120115 TI - Characterization of arginine decarboxylase from Dianthus caryophyllus. AB - Arginine decarboxylase (ADC, EC 4.1.1.9) is a key enzyme in the biosynthesis of polyamines in higher plants, whereas ornithine decarboxylase represents the sole pathway of polyamine biosynthesis in animals. Previously, we characterized a genomic clone from Dianthus caryophyllus, in which the deduced polypeptide of ADC was 725 amino acids with a molecular mass of 78 kDa. In the present study, the ADC gene was subcloned into the pGEX4T1 expression vector in combination with glutathione S-transferase (GST). The fusion protein GST-ADC was water-soluble and thus was purified by sequential GSTrap-arginine affinity chromatography. A thrombin-mediated on-column cleavage reaction was employed to release free ADC from GST. Hiload superdex gel filtration FPLC was then used to obtain a highly purified ADC. The identity of the ADC was confirmed by immunoblot analysis, and its specific activity with respect to (14)C-arginine decarboxylation reaction was determined to be 0.9 CO(2) pkat mg(-1) protein. K(m) and V(max) of the reaction between ADC and the substrate were 0.077 +/- 0.001 mM and 6.0 +/- 0.6 pkat mg(-1) protein, respectively. ADC activity was reduced by 70% in the presence of 0.1 mM Cu(2+) or CO(2+), but was only marginally affected by Mg(2+), or Ca(2+) at the same concentration. Moreover, spermine at 1 mM significantly reduced its activity by 30%. PMID- 15120116 TI - Fructan metabolising enzymes in rhizophores of Vernonia herbacea upon excision of aerial organs. AB - The activities of fructan metabolising enzymes and fructan contents are reported for rhizophores of Vernonia herbacea (Vell.) Rusby induced to sprouting by shoot excision. The activities of fructan exohydrolase (1-FEH), sucrose: sucrose fructosyltransferase (1-SST), fructan: fructan fructosyltransferase (1-FFT) and invertase (INV) and the fructan contents were analysed every 3-4 days for 1 month by colorimetric and chromatographic methods. Sprouting of new shoots started on day 9. 1-FEH activity increased after day 13 and reached its maximum value 20 days after shoot excision. A gradual decrease in 1-SST activity was detected between days 3 and 9. 1-FFT activity exhibited fluctuations throughout the experimental period and a peak of activity for invertase was detected 9 days after shoot excision. Variation in fructan contents in vivo included a decrease until day 13 after which, levels remained practically unchanged. Fructan depolymerization and sprouting are concomitant processes in V. herbacea and can be induced by shoot excision at any phenological phase. 1-FEH and 1-FFT seemed to act in a concerted way to catalyse fructan depolymerization, while 1-SST was inhibited, possibly due to interruption of sucrose supply to rhizophores from the aerial organs. PMID- 15120117 TI - NTZIP antisense plants show reduced chlorophyll levels. AB - We have isolated and characterized a new photosynthetic tissue-specific gene NTZIP (Nicotiana tabacum leucine zipper) from tobacco (N. tabacum). Its deduced amino acid sequence has two highly conserved regions, leucine zipper and [EX(n)DEXRH](2) motifs, which are related to the gene's biochemical functions. NTZIP was expressed in leaves and stems, but was not detected in roots or flowers, suggesting that its physiological functions might be associated with photosynthesis. Northern blot analysis showed that NTZIP mRNA accumulation was induced by light signals, increased greatly under low temperatures and was repressed by strong light illumination. Furthermore, a number of homologs of NTZIP were isolated from cucumber (Cucumis sativus), rape (Brassica napus), clover (Trifolium repens), willow (Salix babylonica), rosebush (Rusa dovurica), wheat (Triticum aestivum) and spinach (Spinacia oleracea), proving the ubiquitous existence of the NTZIP-like genes in higher plants. Transgenic tobaccos constitutively expressing antisense RNA to NTZIP displayed chlorosis and a lack of ability to turn green even under normal growth conditions. The chlorophyll deficiency was further confirmed by chlorophyll content determination and gas exchange analysis. Based on these observations, we propose that NTZIP may be involved in chlorophyll biosynthesis, and might define a novel family of evolutionarily conserved proteins with its homologs in other plant species. PMID- 15120118 TI - The role of lipoic acid in the regulation of the redox status of wheat irrigated with 20% sea water. AB - The effect of irrigation with 20% sea water was studied in 14 and 21-day-old seedlings of durum wheat (Triticum durum, cv. Ofanto). Comparisons between control (Hoagland's 2 solution) and treated (20% sea water in Hoagland's solution) plants included, besides HPLC determination of reduced (DHLA) and oxidised (LA) forms of lipoic acid, ascorbate and glutathione contents, their redox status, the activity of ascorbate peroxidase (APX, EC 1.11.1.11.) and glutathione reductase (GR, EC 1.6.4.2.). The results indicated a more relevant presence of lipoic acid in the roots in comparison to the shoots. An involvement of its reduced form in the regeneration of the reduced glutathione, at least at 14 days of treatment, suggested, besides its role as dehydrogenase enzyme cofactor, a role in the recycling of the other antioxidants. The amount of LA always increased with growth in shoots and decreased in roots, while DHLA remained constant in control and increased in treated plants. Besides, the oxidised form always decreased with sea water while the reduced form decreased in shoots and increased in roots. The ascorbate pool exerted its positive influence especially in the shoots, while APX and GR activities resulted differently modulated by the salinity level. PMID- 15120119 TI - Hormonal changes throughout maturation and ageing in Pinus pinea. AB - Phytohormones, which are responsible for certain age-related changes in plants, play a major role throughout maturation and ageing. Previous results dealing with this topic allowed us to describe an ageing and vigour index in Pinus radiata based on a ratio between different forms of cytokinins (Cks). The aim of the present study was to extend the studies on the changes in the hormonal status throughout maturation and ageing to Stone pine (Pinus pinea L.). With this aim in mind, a number of Cks were analysed in addition to indole-3-acetic acid (IAA) and abscisic acid (ABA) in terminal buds, axillary buds and in the apical portion of needles collected from trees at different stages of development. The results showed an increasing pattern in the levels of various Cks similar to that found in previous studies on P. radiata. Although the maintenance of the same ratio as an ageing and vigour index was not ratified, these results seem to point to Cks as major hormones throughout maturation and related processes in conifers. The distribution of hormones between the two parts of the needle is also discussed. PMID- 15120120 TI - Effect of red and blue light on the timing of cyclin-dependent kinase activity and the timing of cell division in Chlamydomonas reinhardtii. AB - In this study, we describe the effect of red and blue light on the timing of cell division, DNA synthesis, and activity and presence of cyclin-dependent kinases (CDKs), in synchronous cultures of the unicellular green alga Chlamydomonas reinhardtii. Cell division and DNA synthesis were found to occur later in cells grown in blue or white light, than in red light. CDK-like activity, measured using a histone H1 kinase assay, correspondingly occurred later in cultures that were grown in blue light compared to cultures grown in red light. The amount of CDK-like proteins, as detected using an antibody against the PSTAIRE motif, showed a maximum during the division phase. We conclude that the mechanism that causes the delay in the timing of cell division in blue light has its action before DNA replication takes place and also precedes the increase in CDK-like activity. PMID- 15120121 TI - Compulsory winding in the opposite direction of climbing plants promotes yield. AB - The stem of kidney bean plant (Phaseolus vulgaris L., cv. Kentucky 101), a typical dextrorse climbing plant, was subjected to compulsorily sinistrorse winding. The compulsory sinistrorse-winding induced changes in physiological activities. The number of pods with immature seeds (used as vegetable) was doubled and the fresh weight of the pods also significantly increased by sinistrorse-winding. Compulsory sinistrorse-winding increased chlorophyll content, photosynthetic rate, respiration, nodule formation, N(2)-fixation, glutamine synthetase [L-glutamate: ammonia ligase (ADP-forming); E.C. 6.3.1.2] activity and protein content. Thus, it seems to affect the basic physiological processes that promote physiological activities though the action mechanism is unknown. PMID- 15120123 TI - Radiation damage in the TEM and SEM. AB - We review the various ways in which an electron beam can adversely affect an organic or inorganic sample during examination in an electron microscope. The effects considered are: heating, electrostatic charging, ionization damage (radiolysis), displacement damage, sputtering and hydrocarbon contamination. In each case, strategies to minimise the damage are identified. In the light of recent experimental evidence, we re-examine two common assumptions: that the amount of radiation damage is proportional to the electron dose and is independent of beam diameter; and that the extent of the damage is proportional to the amount of energy deposited in the specimen. PMID- 15120124 TI - Charge density determination in icosahedral AlPdMn quasicrystal using quantitative convergent beam electron diffraction. AB - Charge density distribution in icosahedral AlPdMn quasicrystal has been studied on a single-crystal specimen by using quantitative convergent beam electron diffraction (QCBED) technique. The QCBED systematic row method was used in the refinement of structure factors. To refine the low-order structure factors, the wave-mechanical formulation of electron diffraction dynamical theory was used in the calculation of electron diffraction intensities for the quasicrystal in fitting the experimental intensity line scan profiles. The shapes of atomic surfaces (occupation domains) were described with symmetry-adapted series of surface harmonics. An iterative procedure was used in determination of structure factors of the quasicrystal. The structure factors of nine strongest symmetry inequivalent reflections according to X-ray diffraction experiment were refined with QCBED technique. The average of refinement results for a given reflection performed on several CBED patterns, which were slightly different in orientation and sample thickness, and on different line scans, was taken as the value of structure factor for the reflection. The obtained structure factors for electrons were transformed into X-ray structure factors with Mott formula. The bonding charge density map for the quasicrystal was constructed with the obtained nine structure factors. Assuming that the atoms are spheres, the gain or loss of electrons for different atoms were calculated. It shows that identical atoms can have different valences at different kinds of positions. The bonding charge is localized along certain directions. PMID- 15120125 TI - Structural properties and charge ordered states in RMnO3 (R=La, Pr, Nd, Ca, Sr) and (La, Sr)2NiO4. AB - Structural distortions arising from the condensations of two essential kinds of phonon modes: the triply degenerate rotational modes (phix, phiy, phiz) of MnO(6) and the doubly degenerate Jahn-Teller active modes (Q1, Q2) have been systematically investigated in the perovskite manganites. Microstructural features associated with certain types of distortions have been observed by transmission electron microscopy (TEM). In RMnO(3) and La(Sr)(2)NiO(4), we characterize the local structure, charge ordered states and orbital ordering by means of low-temperature TEM. We present direct evidence that the stripe modulation in La(Sr)(2)NiO(4) is indeed one-dimensional within each NiO(2) plane. Several typical kinds of defect structures, including antiphase boundaries and the 90 degrees -twin domains, appear commonly in the charge-ordered states. PMID- 15120126 TI - How to optimize the experimental design of quantitative atomic resolution TEM experiments? AB - A quantitative measure is proposed to evaluate and optimize the design of quantitative atomic resolution TEM experiments. It aims at precise measurement of unknown structure parameters. Specifically, the proposed measure quantifies the statistical precision with which positions of atom columns can be estimated. The optimal design is then given by the combination of microscope settings for which this precision is highest. The proposed measure is also used to find out if new instrumental developments improve the precision as compared to existing methods. PMID- 15120127 TI - Differential-aperture X-ray structural microscopy: a submicron-resolution three dimensional probe of local microstructure and strain. AB - A recently developed differential-aperture X-ray microscopy (DAXM) technique provides local structure and crystallographic orientation with submicron spatial resolution in three-dimensions; it further provides angular precision of approximately 0.01 degrees and local elastic strain with an accuracy of approximately 1.0 x 10(-4) using microbeams from high brilliance third generation synchrotron X-ray sources. DAXM is a powerful tool for inter- and intra-granular studies of lattice distortions and lattice rotations on mesoscopic length scales of tenths of microns to hundreds of microns that are largely above the range of traditional electron microscopy probes. Nondestructive, point-to-point, spatially resolved measurements of local lattice orientations in bulk materials provide direct information on geometrically necessary dislocation density distributions through measurements of the lattice curvature in plastically deformed materials. This paper reviews the DAXM measurement technique and discusses recent demonstrations of DAXM capabilities for measurements of microtexture, local elastic strain, and plastic deformation microstructure. PMID- 15120128 TI - Domain structures in rutile in ultrahigh-pressure metamorphic rocks from Dabie Mountains, China. AB - According to the HRTEM study, the UHP jadeite-quartzite mineral (Rutile, TiO(2)) in Anhui Province, Dabie Mountains, China, has ultrastructures such as 011 two dimensional commensurable modulated structures or superstructures, [011] twin domain structures, dislocations and crystal deformations. The SAED patterns and HRTEM images indicate the existence of the deformations and stacking faults on the interface of [011] twin crystal of rutile and its two-dimensional commensurate modulated structures with repetition period 0.753 nm (3d(011)) has tetragonal symmetry, cell parameters a = 3a0 = 1.377 nm (a0 = 0.459 nm), c = c0 = 0.3 nm. The modulated structures of rutile were probably caused by the isomorphic replacement of Ti(4+) and position modulation or occupation modulation of oxygen atoms in different degree; the deformation structures reveal that during the process of crystallization and mineralization, this mineral may be affected by the geological environment (such as temperature, pressure and stress), metamorphism and deformation. PMID- 15120129 TI - Electron microscopy investigation of gallium oxide micro/nanowire structures synthesized via vapor phase growth. AB - Large-scale micro/nanosized Ga(2)O(3) structures were synthesized via a simple vapor p9hase growth method. The morphology of the as-grown structures varied from aligned arrays of smooth nano/microscale wires to composite and complex microdendrites. We present evidence that the formation of the observed structure depends strongly on its position relative to the source materials (the concentration distribution) and on the growth temperature. A growth model is proposed, based on the vapor-solid (VS) mechanism, which can explain the observed morphologies. PMID- 15120130 TI - TEM investigation on the growth mechanism of carbon nanotubes synthesized by hot filament chemical vapor deposition. AB - Carbon nanotubes (CNTs) were synthesized using hot-filament chemical vapor deposition on Ni film-coated Si substrate. The CNTs were well-aligned perpendicular to the substrate. The as-grown CNTs were bamboo-like in their morphology, and were investigated using SEM and high-resolution transmission electron microscopy (HRTEM). The SEM and HRTEM studies show that the both ends of a CNT contain metallic catalytic particles, which is different from results previously reported. Our analysis results provide strong evidence that the metallic catalyst remains in a liquid state during nanotube growth. The upward growth pulling force of the CNT layer elongates the liquid nanoparticles, which are finally broken into two parts. One part remains at the substrate surface (base of the CNTs) and is responsible for the catalytic growth of the CNTs. The other part is enclosed at the tip of the CNTs and is inactive during CNT growth. PMID- 15120131 TI - Synthesis and growth mechanism of carbon nanotubes and nanofibers from ethanol flames. AB - The ethanol flame was successfully used to synthesize highly graphitic hollow cored carbon nanotubes (CNTs) and novel disorder solid-cored carbon nanofibers (CNFs). Their morphologies were characterized by using scanning electron microscopy, transmission electron microscopy, high-resolution transmission electron microscopy and Raman spectroscopy. It was found that the mixture of CNTs and CNFs were grown on Ni-contained substrates, whereas only the CNFs were produced on carbon steel and low alloy steel substrates. It has been established that Ni and its compounds play a key role in CNTs growth and Fe and its compounds in CNFs growth. The models of 'hollow-cored mechanism' and 'solid-cored mechanism' were proposed to explain the present CNTs and CNFs formations, based on the theory that 'Fe has a strong affinity for carbon and Ni has a weak affinity for carbon'. It is expected that the present ethanol flame may provide a much simpler and more economic approach for mass-production of CNTs and CNFs by using large flame or multi-flames. PMID- 15120132 TI - Structural properties of silver nanorods with fivefold symmetry. AB - The microstructural features of silver nanorods with the average length of 50 microm and diameters of around 100 nm have been investigated by means of SEM, X ray diffraction, and transmission-electron microscopy (TEM). Silver nanorods in general have a pentagonal shape with a remarkable fivefold symmetry as revealed in cross-section observations. The fivefold axis, i.e. the growth direction, normally goes along the [110]-zone axis direction of the basic fcc Ag-structure. The twinning relationships and relevant twin boundaries among the five subunits in the pentagonal nanorod have been examined by high-resolution TEM and selected area electron diffraction. Defects and stacking faults in this kind of nanorods have been briefly analyzed. PMID- 15120133 TI - Microstructural and compositional characteristics of GaN films grown on a ZnO buffered Si (111) wafer. AB - Polycrystalline GaN thin films have been deposited epitaxially on a ZnO-buffered (111)-oriented Si substrate by molecular beam epitaxy. The microstructural and compositional characteristics of the films were studied by analytical transmission electron microscopy (TEM). A SiO(2) amorphous layer about 3.5 nm in thickness between the Si/ZnO interface has been identified by means of spatially resolved electron energy loss spectroscopy. Cross-sectional and plan-view TEM investigations reveal (GaN/ZnO/SiO(2)/Si) layers exhibiting definite a crystallographic relationship: [111](Si)//[111](ZnO)//[0001](GaN) along the epitaxy direction. GaN films are polycrystalline with nanoscale grains ( approximately 100 nm in size) grown along [0001] direction with about 20 degrees between the (1l00) planes of adjacent grains. A three-dimensional growth mode for the buffer layer and the film is proposed to explain the formation of the as grown polycrystalline GaN films and the functionality of the buffer layer. PMID- 15120134 TI - TEM investigations on ZnO nanobelts synthesized via a vapor phase growth. AB - Unusual ZnO nanostructures have been successfully synthesized via selenium controlled chemical vapor phase growth on Si (111) substrates at about 500 degrees C. The microstructure and chemical compositional characteristics of the ZnO nanomaterials have been systematically investigated by means of analytical transmission electron microscopy (TEM), including energy-dispersive X-ray spectroscopy and electron energy-loss spectroscopy. Most of the nanostructures have a belt-like morphology with typical widths of approximately 150 nm and lengths up to several micrometers. All the investigated materials are found to be stoichiometric ZnO with a hexagonal crystal structure. The growth directions for the nanobelts are found to be [1010] and [2110] respectively. Regular-triangle and needle-like heads with diameters only approximately 25-35 nm have been found in the straight nanobelts. High-resolution TEM images indicate that all the nanostructures are single crystals and free of defects. The growth mechanisms of such interesting and unique morphologies are briefly discussed. PMID- 15120135 TI - Formation and microstructural investigation of Ag-Cu alloy nanoclusters embedded in SiO2 formed by sequential ion implantation. AB - We report on the formation of alloy nanoclusters in silica by Ag/Cu ion sequential implantation. The formation of alloy nanocluster has been evidenced by optical absorption spectra, selected area electron diffraction and energy dispersive X-ray spectra. The microstructure characters of nanoclusters have been studied by high resolution transmission electron microscopy. The lattice distortion of some nanoclusters has been observed. A model has been given to explain the distortion. Some defects (partial dislocation, stacking faults) have been found in nanoclusters. PMID- 15120136 TI - Performing probe experiments in the SEM. AB - A four nanoprobe system has been installed inside a FEI XL30 F scanning electron microscope (SEM), and shown to be fully compatible with the normal functions of the SEM and also a Gatan cold stage (model C1003, -185-400 degrees C). With some selected examples of applications, we have shown that this nanoprobe system may be used effectively for gripping, moving and manipulating nanoobjects, e.g. carbon nanotubes, setting up electric contacts for electronic measurements, tailoring the structure of the nanoobject by cutting, etc. and even for making unexpected nanostructures, e.g. a nanohook. Applications in other areas have also been speculated, limitations or disadvantages of the current design of the probe system were discussed, and methods for possible improvement were suggested. PMID- 15120137 TI - Specific recognition of bacteria by plant LysM domain receptor kinases. PMID- 15120138 TI - The multiple routes of MHC-I cross-presentation. PMID- 15120139 TI - Uniqueness of the mating system in Cryptococcus neoformans. PMID- 15120140 TI - Have archaeal genes contributed to bacterial virulence? PMID- 15120141 TI - History and new insights into host defense against vaginal candidiasis. PMID- 15120143 TI - Drosophila: a polyvalent model to decipher host-pathogen interactions. PMID- 15120142 TI - Assembly of pili in Gram-positive bacteria. PMID- 15120144 TI - Pneumocystis: unraveling the cloak of obscurity. PMID- 15120145 TI - Urticaria and angioedema: an overview. AB - Persistent or frequent episodes of urticaria are difficult to evaluate and treat. The best test to identify most patients with a specific underlying cause (eg, physical trigger, allergen, systemic disease) likely is the taking of a careful and detailed history and performance of a physical examination by a specialist who is knowledgeable in urticarial disease. Further study of the pathogenesis and treatment of urticaria is crucial. Given the limited efficacy of presently approved antihistamine treatments and the significant side effects of steroids and cyclosporine, there is a pressing need to evaluate other anecdotally supported urticaria treatments in randomized, controlled trials. PMID- 15120146 TI - Autoimmune urticaria. AB - A growing body of evidence shows that at least 40% of patients with unexplained (idiopathic) chronic urticaria have clinically relevant functional autoantibodies to the high-affinity IgE receptor on basophils and mast cells. The term "autoimmune urticaria" is used for this subgroup of patients presenting with continuous ordinary urticaria. This article reviews the evidence for the autoimmune hypothesis and other nonantibody serum histamine-releasing factors in the etiopathogenesis of urticaria; defines autoimmune urticaria; looks at how autoimmune urticaria fits into existing classifications of urticaria; proposes diagnostic criteria that may be useful to the clinician; and reviews the management implications for patients with this subset of chronic disease. PMID- 15120147 TI - Urticarial vasculitis and hypocomplementemic urticarial vasculitis syndrome. AB - Urticarial vasculitis is a clinicopathologic entity in which episodes of urticaria are accompanied by histopathologic features of cutaneous vasculitis. The histopathologic definition of vasculitis varies from report to report. In this article, vasculitis is defined as histopathologic features of blood vessel damage: There should be evidence of leukocytoclasis and vessel wall destruction, which may or may not be accompanied by fibrinoid deposits. Red blood cell extravasation and perivascular inflammatory cell infiltrate also may be present. The extent to which each of these elements must be present has been debated. PMID- 15120148 TI - Chronic urticaria and thyroid disease. AB - Because the association of thyroid disease with pruritus and urticaria dates back more than 50 years, many investigators have commented on the possible link between autoimmune thyroid disease and chronic urticaria. This article summarizes an evolving body of literature linking these two conditions and discusses potential mechanisms as to how they concomitantly occur. Treatment options used to manage and control the urticaria are discussed with a focus on how the thyroid gland may have a role in the possible mechanism leading to chronic urticaria in this setting and how thyroid hormone may contribute to resolution of this condition. PMID- 15120149 TI - Physical urticaria. AB - Physical urticarias are a unique subgroup of chronic urticaria in which patients develop urticaria secondary to environmental stimuli. Common triggers include exercise, temperature changes, cold, heat, pressure, sunlight, vibration, and water. Systemic symptoms have occurred during severe episodes. Physical urticarias are responsible for approximately 20% to 30% of all cases of chronic urticaria. A basic knowledge of these unusual disorders is important for all healthcare providers. This article covers the following types of physical urticarias: dermatographism, cholinergic urticaria, local heat urticaria, exercise-induced anaphylaxis, vibratory angioedema, solar urticaria, and aquagenic urticaria. PMID- 15120150 TI - Delayed pressure urticaria. AB - Delayed pressure urticaria is a mechanical urticaria in which pressure causes whealing. Delayed cutaneous erythema and edema occur in association with marked subcutaneous swelling after the application of a sustained pressure stimulus to the skin. The earliest reports and theories of the pathogenesis of delayed pressure urticaria are summarized. Detailed attention is given to making the diagnosis by taking a history and provoking the lesions. The clinical features and natural history are considered. The effects of the disorder on quality of life are delineated, and management strategies are suggested. PMID- 15120151 TI - The spectrum of acquired and familial cold-induced urticaria/urticaria-like syndromes. AB - Acquired cold urticaria syndromes represent one of the more common forms of physical urticaria. The syndromes are heterogenous, and a diagnostic classification is presented to facilitate collation for future studies. Acquired cold urticaria represents an excellent reproducible in vivo model to investigate the mechanisms of urticaria. The discussion includes clinical manifestations, laboratory features, pathogenesis, and management of these disorders. A description of familial types, particularly familial cold auto-inflammatory syndrome (FCAS) that is manifested by cold-evoked signs and symptoms of chronic inflammation, is included. FCAS historically has been included with acquired cold urticaria, even though the exanthem of FCAS is maculopapular caused by leukocytic infiltration. FCAS has become an important investigative syndrome, as it represents a reproducible in vivo model of chronic inflammation. PMID- 15120152 TI - Urticaria pigmentosa. AB - Urticaria pigmentosa (UP), resulting from the accumulation of excessive numbers of mast cells in the skin, is the most common form of cutaneous mastocytosis. Observations highlight the diversity of this disease. Clonal expansion of early hematopoietic progenitor cells carrying activating mutations in KIT seems to be the basis of adult-onset UP. New pathogenetic findings are leading to the development of new diagnostic surrogate markers of disease and therapeutic approaches targeting neoplastic mast cells. Promising strategies may arise from an increased understanding about the cause of mastocytosis and the signaling pathways initiated by kit activation. PMID- 15120153 TI - Advances in the treatment of chronic urticaria. AB - There have been a number of exciting developments in the treatment of allergic diseases in recent years, but the development of new treatments for urticaria has lagged behind. The standard treatment for chronic urticaria (CU) involves the use of H1 antagonists. A number of small but promising studies have found potential benefit with medications that are used less often. This article reviews the established therapies for CU and the experimental evidence for the use of nonstandard and relatively unknown therapies. The potential usefulness of some of the new allergy medications for the treatment of CU also is discussed. PMID- 15120155 TI - Selective modulation of antioxidant enzyme activities in host tissues during Rickettsia conorii infection. AB - The involvement of oxidative mechanisms in the pathogenesis of rickettsiosis was investigated using infection of C3H/HeN mice with sub-lethal and lethal infectious doses of Rickettsia conorii, the causative agent of Mediterranean spotted fever. Microscopic examination of tissues at 48 and 96 h post-infection revealed characteristic pathologic features and the presence of rickettsiae in the endothelium of infected tissues. Activities of key antioxidant enzymes, namely glutathione peroxidase, glutathione reductase, glucose-6-phosphate dehydrogenase, and superoxide dismutase, at these times exhibited a pattern of differential and selective modulation in brain, lungs, and testes of mice infected with viable organisms, whereas heat-inactivated or sonically disrupted rickettsiae had no effect. Of these, most significant changes were evident in the lungs of infected animals. Adaptive alterations in oxidant-scavenging enzymes occurred in apparent correlation with the dose and duration of infection. Treatment with an antioxidant, alpha-lipoic acid, protected against infection induced oxidative injury via regulation of antioxidant enzyme activities and maintenance of reduced glutathione levels. These results suggest the involvement of regulatory enzymes of glutathione redox and superoxide scavenging systems in the antioxidant response during in vivo infection, the extent of which varies with the titer of viable rickettsiae in different organs of the host. PMID- 15120156 TI - Nitric oxide exerts distinct effects in local and systemic infections with Streptococcus pneumoniae. AB - Nitric oxide (NO) is known to be involved in the immune response against a range of organisms. Little is known about the effects of nitric oxide in pneumococcal infections. We have now investigated the role of nitric oxide in local and systemic infections caused by Streptococcus pneumoniae in NOS2 deficient mice. Although a deficiency in NO does not affect survival of mice during pneumococcal pneumonia, NO does control pneumococcal viability within the lung airways and tissue. Bronchoalveolar lavage fluid (BALF) from NOS2-deficient mice contained significantly elevated TNF activity, IFNgamma and total protein during mid/late infection. Incubation of S. pneumoniae with the NO donor SNAP revealed a direct anti-pneumococcal effect for NO in vitro. Deficiency in NOS2 did not affect bacteraemia following intranasal infection. In contrast NOS2-deficient mice were significantly less susceptible to intravenous infection with S. pneumoniae than were wild type mice and were able to control pneumococcal viability within the bloodstream. Our results indicate that NO is required within the lungs for anti bacterial activity during the pneumococcal pneumonia but during Gram-positive bacteraemia NO is associated with increased bacterial loads and reduced survival. PMID- 15120157 TI - Susceptibility of immunodeficient mice to aerosol and systemic infection with virulent strains of Francisella tularensis. AB - Previous studies have shown that IFN-gamma, TNF-alpha and NOS-2, but not B cells, are crucial for host defense against primary systemic infection with the attenuated live vaccine strain (LVS) of Francisella tularensis. In this study, we examined the importance of these and additional immune components in host resistance against infection with virulent strains of F. tularensis initiated by systemic and airborne routes. Wild-type (WT) mice and mice deficient in IFN gamma, TNFR1R2, NOS-2, or B cells were equally susceptible to low dose ( approximately 10 colony forming units) aerosol or intradermal challenge with virulent type B F. tularensis, and succumbed to the infection between days 6 and 8 post-inoculation. Quantitative bacteriology showed that IFN-gamma-/- and B cell /- mice consistently harbored up to one log(10) more bacteria in their lungs, spleens and livers than WT mice at day 5 post aerosol exposure. Surprisingly, however, compared to other strains of KO mice and WT control mice, IFN-gamma-/- mice showed only mild liver damage as assessed by histopathology and liver function tests. Additional experiments established that even mice with broad immunodeficiency (SCID, neutropenic, splenectomized or thymectomized mice and mice treated with corticosteroid) were no more susceptible to aerosol-initiated infection with virulent type B or type A F. tularensis than immunosufficient control mice. Combined, our results indicate that, unlike LVS, normal type A and type B F. tularensis strains are so extremely virulent that even immunocompetent mice are virtually defenseless to low dose aerosol and intradermal challenges with them. PMID- 15120158 TI - Loss of lipopolysaccharide receptor CD14 from the surface of human macrophage like cells mediated by Porphyromonas gingivalis outer membrane vesicles. AB - Porphyromonas gingivalis, the major etiologic agent of chronic periodontitis, produces a broad spectrum of virulence factors, including outer membrane vesicles. In this study, we investigated the capacity of P. gingivalis vesicles to promote the shedding or cleavage of the lipopolysaccharide (LPS) receptor CD14 from the surface of human U937 macrophage-like cells. SDS-PAGE/Western immunoblotting analysis of gingival crevicular fluid samples from patients affected by moderate or advanced periodontitis revealed the presence of soluble CD14 and CD14 fragments, thus supporting the hypothesis of an in vivo shedding and cleavage of CD14 receptors. Flow cytometry analysis of macrophage-like cells treated with a vesicle-containing culture supernatant of P. gingivalis showed a significant decrease in the binding of anti-human CD14 to the cell surface. However, no accumulation of soluble CD14 or immunoreactive CD14 fragments in the assay supernatant could be demonstrated by ELISA. Treatment of macrophage-like cells with various concentrations of P. gingivalis vesicles substantially suppressed TNF-alpha production triggered by Escherichia coli LPS. This suppressive effect was much less important using heat-treated vesicles or in the presence of leupeptin, a gingipain inhibitor, during the treatment. Recombinant human CD14 receptors were found to be susceptible to proteolytic degradation by P. gingivalis vesicles. A purified Arg-gingipain preparation produced much more degradation than a Lys-gingipain preparation. This study provides evidence that P. gingivalis outer membrane vesicles contribute to the loss of membrane-bound CD14 receptors and that gingipains degrade this LPS receptor. Such a phenomenon, which results in an hyporesponsiveness of macrophages to LPS stimulation, may contribute to an increased capacity of P. gingivalis, and other periodontopathogens, to evade the host immune system mechanisms. PMID- 15120159 TI - Identification and characterisation of hyaluronate lyase from Streptococcus suis. AB - Hyaluronate lyase, which catalyses the degradation of hyaluronic acid (HA), has been described from several pathogenic streptococcal species. We describe, for the first time, identification and purification of hyaluronate lyase from the zoonotic pig pathogen Streptococcus suis. We have cloned the hyaluronate lyase gene from S. suis and used it to generate an allelic replacement knock-out mutant of S. suis serotype 7 that can no longer biosynthesise the enzyme. Interestingly, a limited strain survey indicates that hyaluronate lyase activity is not present in all disease isolates of S. suis. Polyclonal anti-hyaluronate lyase anti-serum raised against our recombinant hyaluronate lyase has been used in Western blots, showing that hyaluronate lyase activity is always associated with the presence of protein of the expected size, whereas lack of hyaluronate lyase activity is due to truncation or absence of the enzyme. We show that hyaluronate lyase activity is required for S. suis to use HA polymer as a carbon source and that supplying exogenous recombinant hyaluronate lyase to all S. suis strains tested allowed fermentation of the resultant HA breakdown products. PMID- 15120160 TI - Differences in the carriage and the ability to utilize the serotype associated virulence plasmid in strains of Salmonella enterica serotype Typhimurium investigated by use of a self-transferable virulence plasmid, pOG669. AB - Most strains of Salmonella enterica subspecies enterica serotype typhimurium (S. typhimurium) naturally harbour a virulence plasmid which carries the salmonella plasmid virulence (spv) genes. However, isolates belonging to certain phage types are generally found without the plasmid. We have utilized a self-transferable virulence plasmid, pOG669 to investigate the effect of introduction of spv genes into strains of such phage types. The use of the co-integrate plasmid, pOG669, was validated on a diverse collection of strains. pOG669 was transferred into strains of serotypes that are normally associated with the possession of virulence plasmids. All strains maintained the wild type level of virulence in a mouse model, except that introduction of pOG669 restored normal virulence levels in an avirulent, plasmid free strain of S. dublin and resulted in a decrease in virulence in a strain of S. dublin from clonal line Du3. S. gallinarum did not become virulent in mice, but pOG669 was functionally interchangeable with the wild type plasmid when strains were tested in a chicken model. Strains of serotypes not normally associated with the carriage of a virulence plasmid did not increase in virulence upon the introduction of pOG669. An IncX plasmid pOG670 that was included as control was incompatible with the virulence plasmid in a strain of S. dublin, demonstrating that the common virulence plasmid of this serotype is of a different incompatibility group than other virulence plasmids. Strains of S. typhimurium from phage types that do not normally carry a virulence plasmid responded differently to attempts to introduce pOG669. No transconjugants were observed with the strains of DT5 and DT21. The introduction of pOG669 did not alter the virulence of JEO3942(DT10), DT35 and JEO3949(DT66) significantly, while DT1 and DT27 became more virulent. DT27 became as virulent as wild type C5, while logVC(10) of DT1 only increased from 4.1 to 5.7. The ability to express spv genes was measured by use of an spvRAB'-cat fusion. Expression in S. enteritidis was found to be higher than in other serotypes tested. Only serotypes that naturally carry a virulence plasmid expressed spv-genes. The strain of DT1 expressed spv at a very low level, while expression in the strains of DT10 and DT35 was approximately 2-fold lower than in a control strain of S. typhimurium, while the level in the DT66 strain corresponded to the control strain. The plasmid pSTF9, which carried the fusion gene could not be introduced into the strains of DT5, DT21 and DT27. The RpoS level in the strains was measured indirectly by use of a katE-lacZ fusion. In the DT5 strain the level of expression was low, while the strains JEO3942(DT10), DT21, DT27 and DT35 expressed 4-5 fold the level in this strain. An internal fragment of the rpoS gene was sequenced in three strains. These all showed an identical sequence to a published S. typhimurium rpoS gene. PMID- 15120161 TI - Culture of C. burnetii from the dental pulp of experimentally infected guinea pigs. AB - An experimental model of Q fever in Guinea pigs was studied. Coxiella burnetii was cultured from the dental pulp of infected animals following bacteremia. PMID- 15120162 TI - When more means less: a paradox BOLD response in human visual cortex. AB - The predictions of the 'Linear Transfer Model' (LTM) have been tested only by modulating the frequency of the action potentials while keeping the size of the activated neuronal population constant. The LTM states that the blood oxygenation level-dependent contrast (BOLD) signal is directly proportional to the neuronal activity averaged over milliseconds or seconds. We examined the influence on the BOLD response, of manipulating the size of the activated neuronal population while maintaining the electrical discharge activity constant. We performed functional MR measurements on 30 awake, healthy adult volunteers (15 male and 15 female) using a flashed and reversing checkerboard. These stimuli induced the same vascular response and the same increase in the electrical discharge activity but varied in the size of the neuronal population being activated. The BOLD response measured by the extent of activation and the BOLD signal amplitude, was larger for the flashed than to the reversing checkerboard. An assessment of the local deoxyhemoglobin (HbR) concentration indicated that the neuronal activity was lower during the flashed checkerboard than the reversing checkerboard. Because the checkerboard associated with the lower neuronal activity yielded the larger number of activated voxels and the larger BOLD signal, our results run contrary to the predictions of the 'Linear Transfer Model' and for this reason we refer to them as paradoxical. Stimuli defined by luminance contrast or a chromatic contrast yielded identical results. We conclude that the 'LTM' may apply to stimuli that modulate the electrical discharge activity but not to stimuli that modulate the size of the activated neuronal population. PMID- 15120163 TI - Correlation between temporal response of fMRI and fast reaction time in a language task. AB - Correlation between behavioral parameters and fMRI responses can provide an advanced understanding of the neuronal processes. A lexical decision task was employed to examine the correlation between the reaction time (RT) and the temporal parameters in event-related BOLD responses. Word frequency was manipulated in the experiment. RTs for high-frequency, low-frequency and pseudowords were measured during fMRI (417 +/- 9 ms, 631 +/- 22 ms and 658 +/- 15 ms, respectively). For high-frequency words, RTs were significantly shorter than that for low-frequency and pseudowords (p < 0.0005). In the left inferior frontal region, the FWHM of the fMRI responses was significantly correlated with RT (p < 0.001), which may correspond to areas with sustained activation during the whole processing. PMID- 15120164 TI - A newborn piglet study of moderate hypoxic-ischemic brain injury by 1H-MRS and MRI. AB - Cerebral hypoxia-ischemia (HI) is an important cause of perinatal brain damage in the term newborn. The areas most affected are the parasagittal regions of the cerebral cortex and, in severe situations, the basal ganglia. The aim of this study was to show that the newborn piglet model can be used to produce neuropathology resulting from moderate HI insult and to monitor damage for 7 days. Two acute cerebral HI were induced in newborn Large White piglets by reducing the inspired oxygen fraction to 4% and occluding the carotid arteries. Newborn piglets were resuscitated, extubated and monitored for 7 days. (31)P magnetic resonance spectroscopy (MRS) offers the ability to monitor the severity of the HI insults. Lactate (Lac) was detected in the HI group at 2 h, 3 days and 5 days after insult by (1)H MRS. Lac/n-acetylaspartate and Lac/choline and Lac/creatine ratios increased significantly (p < 0.01) in the HI group 2 h after HI insults and remained high over 7 days. For the HI group, mean T(2) values increased significantly in the parietal white matter (subcortical) for 5 days after HI insult [117.5 (+/-7.4) to 158.5 (+/-19.2) at T+3 days, 167.7 (+/-15.4) at T+5 days and 160.9 (+/-10.1) at T+7 days (p < 0.01)]. This newborn piglet model of moderate HI brain injury with reproducible cerebral damage could be use as reference for the study of neuroprotective strategy for a period of 7 days. PMID- 15120165 TI - A modified logistic model to describe gadolinium kinetics in breast tumors. AB - A five-parameter modified logistic equation is presented that describes the signal enhancement in magnetic resonance dynamic contrast enhanced imaging (MRI DCE). In this heuristic model, P(1) approximates the baseline signal, P(2) is related to the magnitude of the peak signal enhancement, P(3) is the approximate time of the maximum rate of increase of signal, P(4) is related to the maximum rate of signal enhancement, and P(5) is the terminal slope of the signal enhancement curve. Six breast tumors were studied that exhibited diverse patterns of signal enhancement, and in each case, estimated model parameters were well identified. Three of the model parameters, P(2), P(4) and P(5) describe attributes of the signal enhancement curve that have previously been shown to have diagnostic value with respect to breast cancer. Procedures for using the primary model parameters to derive a number of secondary parameters that may also have diagnostic value are discussed. Sensitivity analysis shows that the signal enhancement curve is highly sensitive to P(3) in the region of the signal intensity curve associated with rapid uptake of the contrast reagent. Consequently, frequent signal sampling in this time domain is indicated to enable identification of P(3) and sensitive fitting of the signal intensity curve. The advantages of this heuristic model compared to commonly used compartmental modeling approaches are discussed. PMID- 15120166 TI - Magnetic resonance imaging contrast-enhanced relaxometry of breast tumors: an MRI multicenter investigation concerning 100 patients. AB - Dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) using extracellular contrast agents has proved to be useful for the characterization of breast tumors. DCE-MRI has demonstrated a high sensitivity (around 95%) but a rather poor and controversial specificity, varying, according to the different studies, from 45% to 90%. In order to increase (a) the specificity and (b) the robustness of this quantitative approach in multicenter evaluation (five MRI units), a quantitative approach called dynamic relaxometry has been developed. According to the proposed method, the time-dependent longitudinal relaxation rate measured on region of interest of the lesion was calculated during the contrast uptake, after intravenous bolus injection of contrast agent. A specifically developed method was used for fast R(1) measurements. Relaxometry time curves are fitted to the Tofts model allowing the measurement of the parameters describing the enhancement curve (maximum relation rate enhancement, initial, 30-s and 60-s slopes) and the tissue parameters [transfer constant (K(trans) min(-1)) and extracellular extravascular space fraction (v(e))]. Correspondence factorial analysis followed by hierarchical ascendant classification are then performed on the different parameters. Higher K(trans) values were observed in infiltrative ductal carcinomas than in infiltrative lobular carcinomas, in agreement with data published by other groups. Specificity of DCE-MRI has been increased up to 85%, with a sensitivity of 95% with K(trans)/v(e) and enhancement index I (ratio of initial slope by maximum relaxation rate enhancement). A multiparametric data analysis of the calculated parameters opens the way to include quantitative image based information in new nosologic approaches to breast tumors. PMID- 15120167 TI - Encapsulation of gadobutrol in AVE-based liposomal carriers for MR detectability. AB - Artificial virus-like envelopes (AVEs) are liposomal carriers that may be useful for target-site-specific delivery of contrast agents. We speculated that T(1) relaxation times of a suspension of Gadolinium-filled AVEs might be shortened after internalization and lysosomal breakdown. To test this hypothesis we evaluated the T(1) relaxation times of Gadobutrol-containing AVEs before and after degradation in vitro and after receptor-mediated cellular uptake. AVEs were filled with 1 M Gadobutrol (Gadovist; Schering AG, Berlin, Germany) yielding Gd chelate-AVEs. T(1)-relaxation times were calculated using an inversion recovery technique for different concentrations of the liposomal suspension. AVEs were degraded in vitro to mimic the release of the encapsulated Gadolinium in cells and to determine a putative increase of the T(1)-effect. Finally, Gd-chelate-AVEs where equipped with integrin-binding RGD ligands and the T1 relaxation times of these Gd-chelate-RDG-AVEs were determined after cellular uptake into endothelial or melanoma cells. Gadobutrol could be encapsulated into AVEs at a high concentration of 1 M (Gd-chelate-AVEs). The Gd-chelate-AVEs could be visualized by MRI. Concentrations down to 1:4 x 10(3) showed a significant T(1)-shortening effect. The degradation of the liposomes with Triton X-100 resulted in a further reduction down to concentrations of 1:10 x 10(3). In addition, cellular uptakes of Gd-chelate-RGD-AVEs also lead to a significant T(1)-shortening. Our study shows that Gadolinium can be efficiently encapsulated into AVEs and that Gd chelate-AVEs can be detected by MRI T(1)-weighted measurements. The MRI detectability is enhanced by degradation. Gd-chelate-RGD-AVEs can be used to enhance the Gd uptake in cells expressing the alpha(v)beta(3) receptor. PMID- 15120168 TI - Pulmonary resorption of inhaled gadobutrol in an animal model: usage to determine lung diffusion in MRI examinations. AB - The aim of this study was to examine a pulmonary gadobutrol resorption after inhalation. An examination of 16 young swine, which were given different amounts of gadobutrol through inhalation, was performed. A measurement of the signal elevation in peripheral lung tissue as well as in the thoracic aorta was made using a T(1)-weighted FLASH-3D sequence in a 1.5 T magnetic resonance imaging (MRI) machine. Twenty minutes after gadobutrol application, a steep signal elevation in the aorta was reported, as well as a plateau phase after 45 min. The signal elevation in peripheral lung tissue after inhalation increased to a mean of 33.9%. The concurrent signal elevation of the abdominal aorta was 137.4%. The evaluation of an intravascular signal elevation after gadobutrol inhalation opens the possibility to evaluate and obtain lung diffusion with MRI. PMID- 15120169 TI - Transport of contrast agents in contrast-enhanced magnetic resonance angiography. AB - Contrast-enhanced magnetic resonance angiography (CEMRA) often appears to display vessels with good resolution, but it does not adequately visualize the throat of a tight stenosis. Image quality in CEMRA is also governed by the timing of contrast injection and data acquisition. We developed a numerical technique to predict the image appearance of a target vessel by taking into account the passage of contrast agent. Experiments were conducted on a phantom with both centric and linear ordering. An effort to prolong the duration in which the contrast passes by employing a double injection technique gives a wider window of opportunity to acquire quality images. PMID- 15120170 TI - Multispectral analysis of bone lesions in the hands of patients with rheumatoid arthritis. AB - Quantitative measures of rheumatoid arthritis (RA) disease progression can provide valuable tools for evaluation of new treatments during clinical trials. In this study, a novel multispectral (MS) MRI analysis method is presented to quantify changes in bone lesion volume (DeltaBLV) in the hands of RA patients. Image registration and MS analysis were employed to identify MS tissue class transitions between two serial MRI exams. DeltaBLV was determined from MS class transitions between two time points. The following three classifiers were investigated: (a) multivariate Gaussian (MVG), (b) k-nearest neighbor (k-NN), and (c) K-means (KM). Unlike supervised classifiers (MVG, k-NN), KM, an unsupervised classifier, does not require labeled training data, resulting in potentially greater clinical utility. All MS estimates of DeltaBLV were linearly correlated (r(p)) with manual estimates. KM and k-NN estimates also exhibited a significant rank-order correlation (r(s)) with manual estimates. For KM, r(p) = 0.94 p < 0.0001, r(s) = 0.76 p = 0.002; for k-NN, r(p) = 0.86 p = 0.0001, r(s) = 0.69 p = 0.009; and for MVG, r(p) = 0.84 p = 0.0003, r(s) = 0.49 p = 0.09. Temporal classification rates were as follows: for KM, 90.1%; for MVG, 89.5%; and for k NN, 86.7%. KM matched the performance of k-NN, offering strong potential for use in multicenter clinical trials. This study demonstrates that MS tissue class transitions provide a quantitative measure of DeltaBLV. PMID- 15120171 TI - Cardiac phosphorus-31 two-dimensional chemical shift imaging in patients with hereditary hemochromatosis. AB - Hemochromatosis is a hereditary iron overload syndrome characterized by increased iron storage, followed by liver cirrhosis and is often associated with restrictive cardiomyopathy. The purpose of this study was to detect alterations of cardiac high-energy phosphate metabolism in patients with hereditary hemochromatosis (HHC) prior to the development of structural heart diseases. Therefore cardiac phosphorus-31 two-dimensional chemical shift imaging ((31)P 2D CSI) was employed. Twenty-four male patients (mean age 47.2 +/- 12 years) homozygous for the C282Y mutation in the hemochromatosis associated HFE gene and twenty-four male healthy volunteers (mean age 47 +/- 11 years) as age-matched controls were included in this study. Using a 1.5-Tesla whole-body magnetic resonance scanner, electrocardiograph-triggered transversal 31P 2D CSI was performed. Left ventricle mean phosphocreatine (PCr) to beta-adenosine triphosphate (beta-ATP) ratios of patients with HHC (1.60 +/- 0.41) were significantly decreased in comparison to healthy volunteers (1.93 +/- 0.36; p = 0.004). Furthermore, we detected moderate, negative correlations between left ventricular PCr to beta-ATP ratios and transferrin saturation, cholesterol, low density lipoprotein as well as triglyceride. This study shows that 31P 2D CSI permits the detection of alterations of cardiac high-energy phosphate metabolism in patients with HHC, but without any evidence for heart disease. The decreased PCr to beta-ATP ratios in HHC might be caused by mitochondrial impairment due to cardiac iron overload. PMID- 15120172 TI - A comparative study of myo-inositol quantification using LCmodel at 1.5 T and 3.0 T with 3 D 1H proton spectroscopic imaging of the human brain. AB - Myo-inositol is a strongly coupled system and resonates at four chemical shift positions. At 1.5 T, only the singlet component at 3.57 ppm is detected. However, at 3 T this resonance is resolved into its components at 3.55 ppm and 3.61 ppm. Due to the increased spectral resolution and signal-to-noise ratio, it is anticipated that the quantification of myo-inositol should improve at 3 T. Using data from normal controls and the LCmodel quantification procedure, we found that the quantification precision, reproducibility and detection sensitivity of myo inositol is significantly better at 3 T relative to 1.5 T. PMID- 15120173 TI - A novel phantom and method for comprehensive 3-dimensional measurement and correction of geometric distortion in magnetic resonance imaging. AB - A phantom that can be used for mapping geometric distortion in magnetic resonance imaging (MRI) is described. This phantom provides an array of densely distributed control points in three-dimensional (3D) space. These points form the basis of a comprehensive measurement method to correct for geometric distortion in MR images arising principally from gradient field non-linearity and magnet field inhomogeneity. The phantom was designed based on the concept that a point in space can be defined using three orthogonal planes. This novel design approach allows for as many control points as desired. Employing this novel design, a highly accurate method has been developed that enables the positions of the control points to be measured to sub-voxel accuracy. The phantom described in this paper was constructed to fit into a body coil of a MRI scanner, (external dimensions of the phantom were: 310 mm x 310 mm x 310 mm), and it contained 10,830 control points. With this phantom, the mean errors in the measured coordinates of the control points were on the order of 0.1 mm or less, which were less than one tenth of the voxel's dimensions of the phantom image. The calculated three-dimensional distortion map, i.e., the differences between the image positions and true positions of the control points, can then be used to compensate for geometric distortion for a full image restoration. It is anticipated that this novel method will have an impact on the applicability of MRI in both clinical and research settings, especially in areas where geometric accuracy is highly required, such as in MR neuro-imaging. PMID- 15120174 TI - Intermolecular double-quantum coherence MR microimaging of pig tail with unique image contrast. AB - Image contrast in intermolecular double-quantum coherence (iDQC) imaging of a pig tail was investigated on a 7.05-T microimaging scanner. In addition to TR (repetition time) and TE (echo time), the time interval tau between radio frequency pulses during iDQC evolution and the areas under the iDQC-encode gradients in the iDQC imaging sequence were also used to manipulate image contrast. When suitable imaging parameters were selected, images with unique contrast, such as those with certain regions of the sample highlighted, were obtained without using contrast agents. The effects of iDQC-encode gradient on image contrast were studied quantitatively, and the unique contrast imposed by the related diffusion weighting was also shown. Experimental results demonstrated that the iDQC images have contrast fundamentally different from the conventional single-quantum coherence images. PMID- 15120175 TI - In vivo observation of oxygen-supersaturated water in the human mouth and stomach. AB - In recent years, a rising number of different table waters supersaturated with oxygen have hit the market with claims of both positive health effects and an increase in athletic performance. A scientific validation of these claims needs additional knowledge on the fate of the oxygen supersaturation in the human digestive tract. Taking advantage of the fact that molecular oxygen is paramagnetic, MRI can be applied to observe the behavior of oxygen-supersaturated water after oral uptake. In this contribution we report results obtained on several healthy volunteers. On the basis of these results we can conclude that oral uptake of oxygen-supersaturated drinking water with a low content in CO(2) leads to a considerable increase in the oxygenation in the lumen of the oral cavity and of the stomach. Comparing the observed contrast changes with those brought about by conventional contrast agents, even the highly oxygen supersaturated waters still perform rather poorly. PMID- 15120176 TI - Field gradient CPMG applied on postmortem muscles. AB - As a new approach, Carr-Purcell-Meiboom-Gill (CPMG) experiments were performed in vitro on porcine muscles (n = 10) during the period from 15 min to 85 min postmortem and again at 24 h postmortem in the absence (G = 0) and the presence of an external field gradient (G = 0.5*10(-3) T/m), which was applied throughout the CPMG sequence. The experiments were performed on low-field nuclear magnetic resonance (NMR) equipment (0.47 T). Due to the inclusion of different pre slaughter treatments (adrenaline treatment and pre-slaughter exercise/electrical stunning), the muscles could be divided into (I) a group (n = 5) characterized by a reduced decrease in pH postmortem and a high water-holding capacity and (II) a group (n = 5) characterized by an increased rate of pH decrease postmortem and a low water-holding capacity. Distributed analysis of the CPMG data revealed two major relaxation populations with relaxation times about 30-40 and 200-500 ms, respectively, and comparison of data obtained with G = 0 and G = 0.5*10(-3) T/m revealed effects of the external gradient on the relaxation time of both the two relaxation populations, which implies that both diffusion and relaxation contributes to the relaxation of the two populations. At 24 h postmortem the effect of the external field gradient on the relaxation time was significantly affected by muscle group (I vs. II), which reveals local differences in water diffusion in the two meat qualities. Finally, the discriminatory power with regard to muscle group (I vs. II) was investigated for data acquired with G = 0 and G = F = 0.5*10(-3) T/m, and both the two types of data were found highly suitable for separation of muscles according to meat quality. PMID- 15120177 TI - Myelin characterization of fetal brain with mono-point estimated T1-maps. AB - Magnetic resonance imaging (MRI) allows non-invasive assessment of fetus brain maturation at the beginning of the third trimester because of its high sensitivity for fat and water content changes accompanying the myelin formation. In this article we propose a new ultra-fast mono point T(1)-map method based on simplified optimized Gradient Echo (GE) two-point method. Results are compared between the two methods and the precision discussed. This quantitative method can be used in clinical routine, as sedation is not needed for patients. PMID- 15120178 TI - MRI with the dipolar interaction refocusing techniques: analysis of the effectiveness for the solid-state polymers. AB - The effectiveness of solid-echo and magic-echo phase-encoding solid-state magnetic resonance imaging methods was tested to determine possible improvement of sensitivity and spatial resolution for investigation of various types of solid polymers. The dipolar interaction refocusing pulse sequences have been used to elongate the possible phase-encoding period and to improve the signal sensitivity. The comparison of both dipolar refocusing techniques with conventional single point imaging method was made. The optimization of the phase encoding time and magnetization recovery periods were performed basing on (1)H spectra and longitudinal relaxation measurements, respectively. The influence of imaging artifacts (intrinsic for each technique) on image quality was investigated. The effectiveness of the artifacts suppression methods was tested. PMID- 15120179 TI - Diffuse abdominal angiomatosis. AB - We present a rare case of diffuse abdominal angiomatosis occurring in a 19-year old male; particular attention is paid to the MR imaging features of this disease process. PMID- 15120180 TI - The Langerhans' cell histiocytosis (eosinophilic granuloma) of the cervical spine: a rare diagnosis of cervical pain. AB - We present the case of a 44-year-old man who complained of cervical pain. He was treated with physiotherapy and analgetics. Because of persistent pain, computed tomography (CT) scan and MRI were performed. They revealed an osteolytic destruction of the fourth cervical vertebra. The patient was treated surgically for removal of the tumor and stabilization of his cervical spine. Histology of the osteolytic material led to the diagnosis of an eosinophilic granuloma of the cervical spine. This case report describes the incidence, clinical significance, background and therapy of an eosinophilic granuloma of the spine. PMID- 15120181 TI - The development of a three-dimensional T1 image calculation program in proportion to the DICOM data of any marketing clinical MRI systems. AB - Contrast media such as gadolinium-diethlene-triamine pentaacetic acid (Gd-DTPA) is used for MRI. Recently there have been some reports about diagnosis using contrast media for the MRI T(1) image for the quantitative evaluation of articular cartilage degeneration. This may be a useful method to evaluate the lesion of the articular cartilage or to confirm therapeutic progress. Whether or not contrast is used, the use of the calculated T(1) image is effective for the quantitative evaluation of the degeneration of knee joint cartilage. However, our system copes with the format of every MRI image even if software of the calculated T(1) image isn't often prepared in the commercial MRI device. So, we developed general purpose image data processing software that can be processed on the preexisting three-dimensional image data processing system. PMID- 15120183 TI - Function and regulation of MHC class II molecules in T-lymphocytes: of mice and men. AB - The main function of major histocompatibility complex (MHC) class II molecules is to present processed antigens, which are derived primarily from exogenous sources, to CD4(+) T-lymphocytes. MHC class II molecules thereby are critical for the initiation of the antigen-specific immune response. Besides antigen presentation, growing evidence is showing that ligation of MHC class II molecules also activates intracellular signaling pathways, frequently leading to apoptosis. Constitutive expression of MHC class II molecules is confined to professional antigen-presenting cells (APC) of the immune system, and in nonprofessional APCs MHC class II molecules can be induced by a variety of immune regulators. Interestingly, activated T cells from many species, with the exception of mice, synthesize and express MHC class II molecules at their cell surface. In this review, we discuss our current knowledge on the transcriptional regulation of MHC class II expression in activated human and mouse T cells, and the contribution of DNA methylation of the T-cell employed class II transactivator promoter III to the MHC class II deficiency of mouse T cells. We also discuss the proposed functions of the activated T cell synthesized and expressed MHC class II molecules, including antigen presentation, T-T cell interactions, and MHC class II-mediated intracellular signaling. PMID- 15120184 TI - Anti-HLA class I antibody-mediated activation of the PI3K/Akt signaling pathway and induction of Bcl-2 and Bcl-xL expression in endothelial cells. AB - Anti-human leukocyte antigen (HLA) antibodies (Ab) have long been implicated in the process of acute and chronic allograft rejection, yet their mechanism(s) of action is not well understood. The aim of this study was to determine whether ligation of HLA class I molecules by anti-HLA Ab on the surface of human endothelial cells (EC) activates the PI3 Kinase (PI3K)/Akt signaling pathway and downstream target proteins of the cell death apparatus. We report that Ab ligation of major histocompatibility complex (MHC) class I molecules on the surface of EC triggers phosphorylation of Akt, PI3K, and recruitment of PI3K and Akt into a signaling unit with focal adhesion kinase. Signaling through class I also stimulated phosphorylation of Bad and upregulated expression of Bcl-2 and Bcl-xL. Pretreatment of EC with the PI3K inhibitor wortmannin blocked class I mediated expression of Bcl-2, but not Bcl-xL, suggesting a role for the PI3K/Akt signaling pathway in regulation of class I-induced Bcl-2 expression. The intracellular events initiated by class I ligation were influenced by the concentration of the anti-HLA Ab with the lowest tested concentrations of Ab stimulating the highest level of Akt phosphorylation, Bcl-xL and Bcl-2 expression. Consistent with the in vitro experiments, analysis of biopsy samples from heart transplant recipients with evidence of Ab-mediated rejection exhibited increased Bcl-2 expression on the vascular endothelium. These results suggest that exposure of the graft endothelium to low concentrations of anti-HLA Ab may promote cell survival by transducing signals resulting in upregulation of cell survival genes. PMID- 15120185 TI - Increase in plasmacytoid and myeloid dendritic cells by progenipoietin-1, a chimeric Flt-3 and G-CSF receptor agonist, in SIV-Infected rhesus macaques. AB - As in HIV-1 infection in humans, SIVsm infection of rhesus macaques causes a slow progressive loss of CD4 T-cells followed by the onset of AIDS. In addition, there is a loss of dendritic cells (DC) in peripheral blood, peripheral lymphoid tissues, and the skin. Increasing the number of CD4 T cells and DC may be an important step in restoring immune competence and thus delay disease progression. Recently, progenipoietins (ProGP), a new family of chimeric Flt3 and G-CSF receptor agonists, were demonstrated to possess the capacity to mobilize hematopoietic progenitor cells in normal rhesus monkeys. In addition, these molecules induced increased numbers of myeloid cells, including dendritic cells, in the blood. Here we demonstrate that SIVsm-infected macaques, treated with ProGP-1, developed increased numbers of both plasmacytoid (CD123+, CD11c-) and myeloid (both CD11b+, CD11c+, and CD123-, CD11c+ subsets) DC and CD4 and CD8 T cells in peripheral blood. Importantly, during treatment, no changes in plasma virus load were observed. After 14 to 20 days of treatment, antibodies were formed against ProGP in all animals. As a consequence, white blood cell levels returned to baseline in several animals. In other animals values only returned to baseline after termination of ProGP treatment. In conclusion, ProGP-1 may be used to generate a transient increase in DC as well as CD4 T-cell numbers, thereby creating a window of opportunity for immunotherapeutic intervention. PMID- 15120186 TI - Regulation of FasL expression in natural killer cells. AB - Fas ligand (FasL)-mediated cytotoxicity is initiated in natural killer (NK) cells through ligation of their activating receptors. The CD16 receptor has been shown to induce FasL expression and cytotoxicity in NK cells. In this study, we made the novel observation that FasL expression was upregulated in NKL cells stimulated through 2B4 and LFA-1 activating receptors, implying a role for FasL mediated cytotoxicity early in the immune response. Coligation with CD94/NKG2A human leukocyte antigen (HLA) class I inhibitory receptor did not block the induced FasL expression; therefore, these opposing pathways appear to function independently. We also showed, however, that FasL-mediated cytotoxicity was downregulated in CD94/NKG2A-expressing LAK cells in response to the HLA-E ligand, suggesting a mechanism by which aberrant cells expressing class I may evade FasL mediated cytotoxicity. Thus we show for the first time that 2B4, LFA-1, and CD94/NKG2A receptors are involved in modulating FasL expression and, therefore, cytotoxicity mediated by NK cells. PMID- 15120187 TI - Umbilical cord blood-naive T cells but not adult blood-naive T cells require HLA class II on antigen-presenting cells for allo-immune activation. AB - Because a relatively low incidence and severity of graft-versus-host disease after umbilical cord blood (UCB) transplantation is observed, we investigated whether T cells from UCB or adult blood (AB) were differentially activated by antigen-presenting cells with or without human leukocyte antigen (HLA)-DR expression. T cells from UCB or AB, or CD45RA(+) naive T cells and CD45RO(+) memory T cells separated from AB, were stimulated with the HLA-DR(+) or HLA-DR(-) cell line AML193. On days 1-3 after stimulation, numbers of interleukin (IL)-2, IL-4, IL-10 or interferon gamma (IFN-gamma)-secreting cells were determined by enzyme-linked immunospot analysis. No IL-4 or IL-10 was produced. AML193-DR(+) cells induced IL-2 and IFN-gamma secretion with slower kinetics and lower levels in UCB T cells than in AB T cells. AML193-DR(+) cells induced comparable IL-2 but higher IFN-gamma secretion in CD45RA(+) T cells from AB than in UCB T cells. AML193-DR(-) cells did not induce IL-2- or IFN-gamma secretion in UCB T cells, but stimulated both CD45RA(+) and CD45RO(+) T cells from AB to secrete IL-2 and IFN-gamma. Thus, not only the absence of memory T cells but also the inability to respond to HLA-DR-negative antigen-presenting cells and the slower kinetics and level of activation found for naive T cells from UCB as compared with AB may partly explain the reduced antirecipient reactivity after UCB transplantation. PMID- 15120188 TI - Functional promoter region polymorphism of the proinflammatory chemokine IL-8 gene associates with Parkinson's disease in the Irish. AB - Parkinson's disease (PD) is one of the most prevalent neurodegenerative disorders and is characterized by the progressive loss of dopamine neurons in the substantia nigra. There is increasing evidence to suggest the inflammatory response of the brain contributes to the pathogenesis of PD. This study investigated the frequency of polymorphism located in the critical promoter region of the proinflammatory cytokine genes: interleukin (IL)-2, IL-6, IL-8, and tumor necrosis factor alpha (TNF-alpha) within a cohort of patients with PD in comparison to a group of healthy elderly individuals. No association was observed for single nucleotide polymorphism in the promoter regions of the IL-2, IL-6, and TNF-alpha genes. The single nucleotide polymorphism in the chemokine IL-8 gene was observed to associate with PD and appeared to be independent of age at onset. This association further supports the theory that the proinflammatory response in the brains of patients with PD plays a role in the pathogenesis of the disease and warrants further investigation into the role of chemokines in the brain, and a more detailed analysis of the genetics involved in the immune response of the brain. PMID- 15120189 TI - Association between TNF-alpha and TGF-beta genotypes in infants and parental history of allergic rhinitis and asthma. AB - The development and expression of allergic rhinitis and asthma may be influenced by the elaboration of specific cytokines. Cytokine genotypes moderate illness severity in a variety of inflammatory disorders. Cytokine genotyping was performed on 124 infants (85% white, 57% male) to determine whether specific cytokine genotypes are associated with a parental history of allergic rhinitis and/or asthma. DNA was extracted from buccal brushings and assayed for tumor necrosis factor alpha (TNF-alpha), interferon gamma (IFN-gamma), interleukin (IL) 6, IL-10, and transforming growth factor (TGF)-beta1 genotypes using polymerase chain reaction-sequence specific primer technology. Outcomes consisted of parental history of allergy and asthma, and results were evaluated by logistic regression. TNF-alpha and TGF-beta genotypes were related to maternal and/or paternal history of allergic rhinitis and asthma, respectively. The frequencies of the genotype associated with high production of TNF-alpha were 41% versus 18% in infants with and without a parental history of allergic rhinitis, respectively (p < 0.01). The frequencies of the genotype associated with low production of TGF beta1 were 14% versus 1% in infants with and without a parental history of asthma, respectively (p < 0.01). There were no associations between IFN-gamma, IL 6, and IL-10 genotypes and any of the outcome parameters. These results suggest a role for TNF-alpha and TGF-beta1 genotypes in the pathogenesis of allergic rhinitis and asthma, respectively. If confirmed by future studies, cytokine genotyping may be a useful tool for identifying at-risk infants who may benefit from the selective use of preventative and/or early intervention treatments for these disorders. PMID- 15120190 TI - Allele and haplotype frequencies for HLA class II (DQA1 and DQB1) loci in patients with celiac disease from Spain. AB - Celiac disease (CD) is a complex and multifactorial disease, defined as a malabsorptive disorder of the small intestine resulting from ingestion of gluten. Genetic susceptibility to CD has been associated with human leukocyte antigen (HLA)-DQ2 heterodimer, encoded by the DQA1*0501 and DQB1*02 genes. However, HLA risk factors do not explain the whole genetic predisposition: not all DQ2 encoding haplotypes confer equal susceptibility to CD. The aim of the present work was to confirm the aforementioned findings in a southern European population. With this purpose, 136 unrelated children diagnosed with CD were typed at the DNA level for HLA-DQA1 and -DQB1 loci. Patients are currently attended at the Donostia Hospital (province of Guipuzcoa, Spain). HLA class II typing was performed by polymerase chain reaction-sequence specific primer procedures. Conspicuous frequencies of the alleles associated with susceptibility to CD were observed (DQA1*0501: 0.592, DQB1*0201: 0.471). Accordingly, the haplotypes DQA1*0501-DQB1*0201 and DQA1*0201-DQB1*0202 revealed a strong linkage disequilibrium (18.84% and 18.75%, respectively) when compared with the Spanish general population. Of the total sample, 93.4% (127 individuals) were carriers of DQ2 heterodimer, either in homozygosis or in heterozygosis. This percentage coincides with figures reported in previous studies, implying the effect of other genes in the development of CD. PMID- 15120192 TI - HLA class II typing in newborns reveals a low frequency of the DRB1*04 allele and a high frequency of DRB1*11 allele in three regions of continental Italy. AB - As part of a longitudinal study aimed at defining the natural history of prediabetic autoimmunity and predicting the risk of future cases of type 1 diabetes, 3607 newborns from three regions of continental Italy (Lombardia, Liguria, and Lazio) were subjected to genetic testing to determine human leukocyte antigen-DRB1 (HLA-DRB1) and -DQB1 allele and phenotype frequencies. Polymerase chain reaction and immobilized sequence-specific oligonucleotide probe assays were used to identify ten DRB1 allele lineages and three DQB1 alleles. No major inter-regional differences emerged in the allelic distribution indicating homogeneous distribution of the HLA DRB1-DQB1 alleles among the three regions analyzed. Comparison of our data with those published for other Caucasian populations reveals that these three regions are characterized by a very low frequency of DRB1*04 (8%) and a high frequency of DRB1*11 (25%). The phenotype frequencies of HLA-DQB1*0302 and DQB1*0602 observed are also lower than those reported for other populations. Furthermore, the DRB1*04-DQB1*0302 haplotype was relatively infrequent in our population (5.3% of the newborns tested). These findings furnish a genetic "portrait" of the populations of the analyzed regions that will be useful not only for investigation of the genetic risk of type 1 diabetes mellitus in Italy but also for studies of other autoimmune diseases related to HLA genotypes. PMID- 15120191 TI - HLA-DRB1 alleles and HLA-DRB1 shared epitopes are markers for juvenile rheumatoid arthritis subgroups in Colombian mestizos. AB - We studied the association of human leukocyte antigen (HLA)-DRB1 and HLA-DQB1 alleles and HLA haplotypes with juvenile rheumatoid arthritis (JRA) in 65 patients and 65 controls from Colombia. The JRA subsets were distinguished on the basis of criteria established by the American College of Rheumatology. Two alleles were associated with protection, HLA-DRB1*1501 (p = 0.002) and HLA DRB1*1402 (p = 0.01). HLA-DRB1*1602 (p = 0.0000002) was associated with susceptibility for systemic JRA and HLA-DRB1*1104 (p = 0.0002) for pauciarticular JRA. Amino acid sequences at residues 70-74 of DRB1 chain shared by HLA-DRB1 alleles (shared epitomes) were also informative. The polyarticular JRA subset revealed association with (70)QRRAA(74), which includes HLA-DRB1*04, 01, and (70)DRRAA(74), which includes DRB1*1601, 1602, 1101, and 1104. Two new findings of interest were the association of the haplotypes DRB1*1104, DQB1*0301(p = 0.0002) with pauciarticular JRA and DRB1*1602, DQB1*0301 (p = 0.0000002) association with systemic JRA. The DRB1 alleles of these two haplotypes share the epitope (70)DRRAA(74)and were associated with both the pauciarticular and the systemic subset of JRA. Our results suggest that studies of disease susceptibility in populations of admixed genetic background should take into account the contribution of different ethnic groups or nationalities in the recruitment of controls and patients studied in order to rule out genetic stratification. PMID- 15120193 TI - A sequencing-based typing method for HLA-DQA1 alleles. AB - Sequencing-based typing (SBT) is the most comprehensive method for characterizing human leukocyte antigen gene polymorphisms. Development of a SBT method for DQA1 is hampered because of a deletion of codon 56 in nearly half of the known DQA1 alleles. Sequence electropherograms of heterozygous samples comprising a deletion allele and a non-deletion allele display misalignment after codon 56 because of a three base-pair shift in the deletion allele. To overcome this problem, we have designed three group-specific primer sets to selectively amplify the deletion alleles from the nondeletion alleles. DNA samples are initially polymerase chain reaction (PCR)-typed using these primer sets along with an internal positive control primer set specific to growth hormone gene 1 (hGH1). The positive group specific PCR reactions were selectively repeated without hGH1 control primers, and the amplicons were used as template in sequencing reactions. The sequence data were analyzed to obtain DQA1 types using ABI MatchTools software as well as the newly available Conexio Genomics Assign SBT Genotyping Software. The method was validated using a panel of reference DNA from the University of California, Los Angeles, International DNA Exchange Program. We conclude that the present SBT method is a technically simple and robust procedure to characterize the sequence polymorphisms in exon 2 of DQA1 gene. PMID- 15120194 TI - Long-term variations of folic acid concentrations in the Northern Adriatic. AB - Folic acid (FA), which belongs to B-group vitamins has recently been recognized as possible nutrient for some phytoplankton species (a diatom Phaeodactylum tricornutum). The beneficial role of FA in human health is widely documented. In some countries food fortification with FA has already become mandatory. FA is also used under commercial name of Dosfolat for the treatment of industrial and domestic wastewaters. This is an exploratory study on seasonal and depth variability of FA concentrations in the Northern Adriatic Sea (station 101-max depth 30 m, under the Po River influence). Seawater samples were analysed in the period from February 1998 to February 2003 based on monthly determinations. Those preliminary findings showed that in the upper layer (10 m) FA concentrations were in the range from 0.1 to 28 nM. In the lower layer (20-30 m) FA concentrations were in the range from 0.1 to 12 nM. It was found that phytoplankton and microorganisms which are responsible for the production of FA in the Northern Adriatic (Station 101) were situated at the depth from 5 to 10 m. The observed water column and seasonal distributions depended on a sensitive balance between the production and the consumption of FA. PMID- 15120195 TI - Speciation of heavy metals in marine sediments from the East China Sea by ICP-MS with sequential extraction. AB - Twelve elements (V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Mo, Sn, Cd and Pb) in 24 sediment samples at eight sites (S1-S8) from the East China Sea were analyzed with the BCR sequential extraction (SE) protocol to obtain the metal distribution patterns in this region. The results showed that the heavy metal pollutions in S4 and S8 were more severe than in other sampling sites, especially Cd and Pb pollution. In the top sediments at S4 and S8, both the total contents and the most dangerous non residual fractions of Cd and Pb were extremely high. More than 90% of the total concentrations of V, Cr, Mo and Sn existed in the residual fraction. Fe, Co, Ni, Cu and Zn mainly (more than 60%) occurred in the residual fraction. While Mn, Pb and Cd dominantly presented in the non-residual fractions in the top sediments. The metal distribution patterns with depth and the correlations between total organic carbon (TOC) and the total Fe-Mn content were also investigated. The results showed that, for most of the elements except Fe, the concentration of elements in fraction A in the top sediments was higher than that in other depth. The similar rule was also found in fraction B but not in fraction C. Besides, the distributions of V, Cd in fraction B and Pb, Cd, Cu in fraction C might be affected by TOC. PMID- 15120196 TI - Transfer of metals from soil to vegetables in an area near a smelter in Nanning, China. AB - A field survey was conducted to investigate the metal contamination in soils and vegetables, and to evaluate the possible health risks to local population through foodchain transfer near a smelter in Nanning, southern China. Contamination levels in soils and vegetables with cadmium (Cd), lead (Pb), zinc (Zn) and copper (Cu) were measured, and transfer factors (TF) from soils to vegetable plants and its health risk (risk index, RI) were calculated accordingly. Results showed that both soils and vegetables from villages 1 and 2 (V1 and V2, 1500 m and 500 m from the smelter) were heavily contaminated, compared to a village 50 km from the smelter. Geometric mean of Cd and Pb concentrations in vegetables for V1 and V2, respectively, were 0.15 and 0.24 mg Cd kg(-1) and 0.45 and 0.38 mg Pb kg(-1) (on fresh weight basis). Oral intake of Cd and Pb through vegetables poses high health risk to local residents. Risk indices for V1 and V2, respectively, were 3.87 and 7.42 for Cd, and 1.44 and 13.5 for Pb. The complexity of metal contamination and their health risks are also discussed. PMID- 15120197 TI - Imposex and butyltin contamination off the Oporto Coast (NW Portugal): a possible effect of the discharge of dredged material. AB - Imposex in the gastropod Hinia reticulata, a bioindicator for tributyltin (TBT) contamination, was investigated in locations at different distances from the Oporto harbour-dredged material discharge site. The degree of imposex found was inversely related to the distance to the discharge site, the highest imposex frequency being found closest to the discharge area. However, the levels of imposex found were low when compared with those reported for H. reticulata populations from moderately TBT-contaminated areas. Despite detectable levels of dibutyltin (DBT) and monobutyltin (MBT) in snail tissues (total butyltins ranging from 84 to 410 ng/g Sn dry wt.), TBT was below detection limit in all snail samples collected in open sea. PMID- 15120198 TI - Phytoremediation of polychlorinated biphenyl-contaminated soils: the rhizosphere effect. AB - The objective in the first phase of this study was to screen alfalfa, flatpea, sericea lespedeza, deertongue, reed canarygrass, switchgrass, and tall fescue for phytoremediation of polychlorinated biphenyl (PCB)-contaminated soil. During the second phase, the focus was rhizosphere characterization to optimize PCB phytoremediation. Aroclor 1248 (PCB) was added to soil at 100 mg x kg(-1) of soil. In the first phase, all of the plant species treatments showed significantly greater PCB biodegradation compared to the unplanted controls and the two most effective species were selected for further study. During the rhizosphere characterization study, soil irradiation did not affect PCB biodegradation, but planting significantly increased PCB biodegradation; 38% or less of the initial PCB was recovered from planted pots, compared to more than 82% from the unplanted control soils. Presence of plants significantly increased the biological activity (microbial counts and enzyme activity) of both irradiated and unirradiated soils. Greater bacterial counts and soil enzyme activity were closely related to higher levels of PCB biodegradation. The data showed that Aroclor 1248 biodegradation in soil seem to be positively influenced by the presence of plants and plant-bacteria interactions. Our results suggested that phytoremediation could be an environmentally friendly alternative for PCB contaminated soils. PMID- 15120199 TI - Organochlorine compounds and heavy metals in the soft tissue of the mussel Mytilus galloprovincialis collected from Lake Faro (Sicily, Italy). AB - Three hundred samples of Mytilus galloprovincialis were collected from five stations (north, south, east, west and centre) of Lake Faro to evaluate the concentrations of organochlorine compounds and heavy metals. Quantitative determinations of organochlorine pesticides and PCBs were made by GC-ECD and confirmed with GC-MS. Concentrations of "essential" (Cu, Se and Zn) and "toxic" (As, Cd, Hg and Pb) metals were determined by an atomic absorption spectrophotometer. The results obtained show the low residue levels of p,p'-DDE in six samples of M. galloprovincialis from southern (7.00-11.00 ng/g w.w. and 148.3-275 ng/g l.w.) and western (7.60-15.37 ng/g w.w. and 126.7-256.2 ng/g l.w.) areas of Lake Faro. No appreciable residues of PCBs were found in any of the samples examined. Zn concentrations (range 11.0-18.5 microg/g w.w.) were higher than Cu (range 188.3-396.0 ng/g w.w.) and Se (range 93.5-288.9 ng/g w.w.) in all areas of origin. Cd (range 41.9-63.8 ng/g w.w.), Pb (range 64.8-93.0 ng/g w.w.) and Hg levels (range 5.7-13.1 ng/g w.w.) showed lower concentrations than permitted MRLs. The As levels were below detection limits for the all mussel samples. In conclusion, the absence of PCBs, the low levels of p,p'-DDE, the concentrations of Cd, Hg and Pb below permitted MRLs in M. galloprovincialis, used as a "biological indicator", show that Lake Faro is not at contamination risk from these contaminants and moreover is free from health problems for the consumer of mussel products. PMID- 15120200 TI - Heavy metal levels and esterase variations between metal-exposed and unexposed duckweed Lemna minor: field and laboratory studies. AB - Environmental homogeneity is being continuously disturbed and affected by artificially introduced loads of chemical toxicants that also include heavy metals. The Tiljala wetlands of the eastern fringe of Calcutta, West Bengal (India) are a virtual sink for the deposition of urban and industrial wastes that get admixed with the aquatic environment. We have selected Lemna minor (duckweed), as a representative of the biota surviving therein for the present study. Concentrations of lead, cadmium, chromium, zinc, copper and mercury in the fronds of Lemna were measured to peep into the range of input of heavy metals in the duckweed subjects. Natural unexposed population of duckweed from a domestic pond in Batanagar area, 24 Parganas, West Bengal (India) was also found to accumulate similar concentrations of these metals when cultured in artificially contaminated water in the laboratory. The exposed individuals also exhibited polymorphism with respect to the loci of esterase, as compared to an unexposed control plants. Therefore, the present study suggests EST variations of L. minor to be a potential biomarker of heavy metal pollution. PMID- 15120201 TI - Use of AMS in the marine environment. AB - In recent years, the field of AMS has expanded into many areas of science. This paper reviews a variety of applications of AMS in the marine environment, focusing particularly on recent developments and applications. Following a brief summary of the three main isotope techniques used in environmental studies: dating, tracing and source identification, a number of applications are considered. Traditional (14)C-dating is no longer the dominant application of AMS measurements, and together with measurements of (10)Be, (26)Al and (36)Cl, much of the research is now directed towards an understanding of global climate change via studies of oceanic circulation, atmospheric processes and past climates by cosmic ray exposure dating. Profiles of long-lived cosmogenic radionuclides in sediments and ice cores, as a function of depth and, thus, age, provide key information on past solar variability, production rate changes and atmospheric transport and deposition mechanisms. Useful paleoclimatic information may be derived from these archives both because deposition is influenced by climate and because solar activity (which influences production) and solar radiance (which influences climate) are correlated. In recent years, emphasis has been put on the development and application of AMS techniques for the measurement of heavier long lived isotopes, including (99)Tc, (129)I, (236)U and other actinide isotopes. AMS combines ultra low detection limits and the possibility to analyse isotope ratios that can be difficult with traditional instruments and has been used in a number of applications on the consequences and uses of releases from nuclear energy. Finally, the use AMS in environmental sciences is expected to expand further in the foreseeable future with long-lived cosmogenic radionuclides contributing to a large body of knowledge on processes involving atmosphere, oceans, ice sheets, biosphere, soils and sediments. PMID- 15120202 TI - Risk perception research: socio-cultural perspectives on the public experience of air pollution. AB - This paper reviews recent work in the field of risk perception research, taking its examples primarily from work relating to air pollution issues. The paper opens with a brief discussion of the psychological literature on risk perception, in order to set out the key insights, criticisms and more recent developments associated with such approaches, before turning to the findings of recent socio cultural analyses of perceptions of air pollution. This account, which considers how social and cultural factors influence the way in which people interpret and make sense of risk, draws linkages with psychological risk perception research, revealing that over the last decade there has been a pronounced degree of convergence between the conclusions being reached across these two (historically disparate) fields of research. The paper concludes by evaluating the relevance of risk perception research for the science and policy of risk assessment and management. PMID- 15120203 TI - Development of coastal recreational water quality standards in the Mediterranean. AB - Concern about adverse health effects arising out of human exposure to polluted seawater through bathing and other forms of aquatic recreation has led to the development of various quality criteria and standards worldwide. Attempts at quantifying health hazards from polluted recreational waters have been made in several Mediterranean countries through the conduction of epidemiological studies aimed at establishing direct correlation between the microbiological quality of the water and health effects on exposed population groups. This article provides an overview on the development of the criteria and standards for coastal recreational waters in the Mediterranean by examining the evolution of the WHO guidelines, the WHO/UNEP interim criteria for bathing waters and the EU Directives. It also provides a brief account of the various quality criteria and standards for coastal recreational waters in current use in all the Mediterranean countries. PMID- 15120206 TI - Research news and notes. PMID- 15120204 TI - The effect of fire on soil organic matter--a review. AB - The extent of the soil organic carbon pool doubles that present in the atmosphere and is about two to three times greater than that accumulated in living organisms in all Earth's terrestrial ecosystems. In such a scenario, one of the several ecological and environmental impacts of fires is that biomass burning is a significant source of greenhouse gases responsible for global warming. Nevertheless, the oxidation of biomass is usually incomplete and a range of pyrolysis compounds and particulate organic matter (OM) in aerosols are produced simultaneously to the thermal modification of pre-existing C forms in soil. These changes lead to the evolution of the OM to "pyromorphic humus", composed by rearranged macromolecular substances of weak colloidal properties and an enhanced resistance against chemical and biological degradation. Hence the occurrence of fires in both undisturbed and agricultural ecosystems may produce long-lasting effects on soils' OM composition and dynamics. Due to the large extent of the C pool in soils, small deviations in the different C forms may also have a significant effect in the global C balance and consequently on climate change. This paper reviews the effect of forest fires on the quantity and quality of soils' OM. It is focused mainly on the most stable pool of soil C; i.e., that having a large residence time, composed of free lipids, colloidal fractions, including humic acids (HA) and fulvic acids (FA), and other resilient forms. The main transformations exerted by fire on soil humus include the accumulation of new particulate C forms highly resistant to oxidation and biological degradation including the so-called "black carbon" (BC). Controversial environmental implications of such processes, specifically in the stabilisation of C in soil and their bearing on the global C cycle are discussed. PMID- 15120207 TI - The things Indians knew first. PMID- 15120208 TI - Aquaporins and brain edema. AB - Aquaporins are a family of transmembrane proteins that selectively allow the passage of water through the plasma membrane. Their importance is highlighted by their ubiquitous presence from bacteria to mammals. In humans, they are found throughout the body and recent work has highlighted their function within the brain. They are intimately involved in the production of cerebrospinal fluid and the control of water movement at the blood-brain barrier. Aquaporin levels are up regulated in animal models of trauma, stroke and water intoxication as well as around human malignant brain tumors. They have thus been implicated in the formation of brain edema. Knockout mice, without the aquaporin gene, appear to have reduced brain edema compared to their wild type brethren in models of brain edema. Currently, the clinical treatment of brain edema is limited. Increased knowledge of the aquaporins may open new targeted therapies for brain edema. PMID- 15120209 TI - Type 2 diabetes mellitus, etiology and possible treatment: preliminary report. AB - BACKGROUND: Insulin resistance has been proposed as the initial step in the cascade toward type 2 diabetes mellitus. The mechanisms underlying the development of insulin resistance are not fully understood. We hypothesize that neurovascular interactions, in particular arterial elongation, causes compression of the right lateral medulla, triggering a state of autonomic dysfunction including hyperactivity of pancreatic endocrine function, and predisposes to insulin resistance and the development of type 2 diabetes. METHODS: The clinical and operative findings were reviewed retrospectively in 15 patients with primary diagnoses of various right-sided cranial rhizopathies, but with a common diagnosis of type 2 diabetes mellitus. After microvascular decompression was performed for the primary diagnosis, arterial compression was observed of the lateral medulla and cranial nerve X and treated with microvascular decompression. Known duration of the diabetes ranged from "new" (patient was diagnosed as a result of preoperative blood work) to 16 years (mean 7.3 years). Duration of diabetes diagnosis was unknown in 2 patients. Follow-up was from 3 to 113 months (mean 29.9 months). RESULTS: Ten of the 15 patients (66%) showed improvement in their blood glucose control; 5 of those 10 (50%) did so with no (4 patients) or less (1 patient) diabetes medication. CONCLUSIONS: We have shown that arterial compression of the right lateral medulla is consistently present in patients with diabetes mellitus and that microvascular decompression can be performed safely. Further studies are necessary and are under way. PMID- 15120215 TI - Giant pituitary tumors: a study based on surgical treatment of 118 cases. AB - BACKGROUND: The aim of the study is to analyze the nature, extensions, and dural relationships of hormonally inactive giant pituitary tumors. The relevance of the anatomic relationships to surgery is analyzed. METHODS: There were 118 cases of hormonally inactive pituitary tumors analyzed with the maximum dimension of more than 4 cm. These cases were surgically treated in our neurosurgical department from 1995 to 2002. Depending on the anatomic extensions and the nature of their meningeal coverings, these tumors were divided into 4 grades. The grades reflected an increasing order of invasiveness of adjacent dural and arachnoidal compartments. The strategy and outcome of surgery and radiotherapy was analyzed for these 4 groups. Average duration of follow-up was 31 months. RESULTS: There were 54 giant pituitary tumors, which remained within the confines of sellar dura and under the diaphragma sellae and did not enter into the compartment of cavernous sinus (Grade I). Transgression of the medial wall and invasion into the compartment of the cavernous sinus (Grade II) was seen in 38 cases. Elevation of the dura of the superior wall of the cavernous sinus and extension of this elevation into various compartments of brain (Grade III) was observed in 24 cases. Supradiaphragmatic-subarachnoid extension (Grade IV) was seen in 2 patients. The majority of patients were treated by transsphenoidal route. CONCLUSIONS: Giant pituitary tumors usually have a meningeal cover and extend into well-defined anatomic pathways. Radical surgery by a transsphenoidal route is indicated and possible in Grade I-III pituitary tumors. Such a strategy offers a reasonable opportunity for recovery in vision and a satisfactory postoperative and long-term outcome. Biopsy of the tumor followed by radiotherapy could be suitable for Grade IV pituitary tumors. PMID- 15120212 TI - ICP threshold in CPP management of severe head injury patients. AB - BACKGROUND: Elevated intracranial pressure (ICP) is significantly associated with high mortality rate in severe head injury (SHI) patients. However, there is no absolute agreement regarding the level at which ICP must be treated. The objective of this study was to compare the outcomes of severe head injury patients treated by setting the ICP threshold at >or=20 mm Hg or >or=25 mm Hg. METHODS: Treatment protocol in this study consisted of therapeutic maneuvers designed to maximize cerebral profusion pressure (CPP) and control ICP. Twenty seven patients with severe head injury and intracranial hypertension (ICP >or=20 mm Hg) were enrolled and fourteen cases were allocated to the group of ICP threshold >or=25 mm Hg. Six-month clinical outcomes were evaluated using the Glasgow Outcome Score (GOS). RESULTS: There were no statistically significant differences in clinical parameters between the groups. Logistic regression identified the presence of basal cisterns on the initial computed tomography (CT) scan as a significant predictor of good outcome. ICP threshold did not influence outcome. CONCLUSIONS: This study supported a recommended ICP threshold of 20 to 25 mm Hg in SHI management. However, in cases with an absence of basal cisterns on initial CT scan, the probability of good outcome may be higher using an ICP threshold of >or=20 mm Hg. PMID- 15120218 TI - Association between Cowden syndrome and Lhermitte-Duclos disease: report of two cases and review of the literature. AB - BACKGROUND: The authors describe 2 cases of dysplasic gangliocytoma of the cerebellum or Lhermitte-Duclos disease revealing Cowden disease or multiple hamartoma neoplasia syndrome. Cowden disease is a rare autosomal dominant disorder, now considered as a phakomatosis. Nevertheless, relationships between both conditions still remain unclear, since Lhermitte-Duclos disease can also be sporadic. CASE REPORTS: Two patients, 25 and 27 years old, were admitted to the emergency department for an acute intracranial hypertension. In both cases, magnetic resonance imaging (MRI) scan showed a mass in the cerebellar hemisphere. Pathological examination of surgical resection specimens concluded Lhermitte Duclos disease. Because of the patients' previous personal and familial medical history, Cowden disease was suspected and confirmed by mutational analysis of the phosphatase and tensin homolog (PTEN) gene. In the first case, a nonsense punctual mutation in exon 8 was found. In the second one, a mutation was revealed in the exon 5, a mutational hot spot encoding the phosphatase catalytic core motif. CONCLUSION: Lhermitte-Duclos disease and Cowden disease can be associated. Germline mutations of PTEN gene are known to be implicated in Cowden disease. This gene, located at chromosome 10q23-3, is a tumor suppressor gene that encodes a protein with phosphatase activity. To date, more than 80 mutations have been reported in Cowden disease. When the diagnosis of either one of these two disorders is established, it is imperative to search for the other one to detect early malignant lesions that occur in Cowden disease. Finally, a long-term follow up of the patient is required and a thorough familial screening is necessary. PMID- 15120220 TI - E-mail consultation: clinical, financial, legal, and ethical implications. AB - BACKGROUND: Communication via e-mail has become widespread. Nearly every practicing neurosurgeon is confronted with numerous unsolicited e-mail requests for medical advice, guidance, or information. Neurosurgeons need to be aware of the clinical, financial, legal, and ethical implications of providing medical consultation via e-mail. METHODS: A literature review of the penetration of e mail consultation in medical practice was performed. The data on the potential for reimbursement for provision of these services is presented. Precedents for legal liability are discussed, and issues of compliance with HIPAA regulations are reviewed. RESULTS: Communication between patients and physicians via e-mail is increasing in prevalence, and a substantial number of physicians are providing medical information via e-mail consultation. Billing for online consultation has been approved by the American Medical Association, and several medical insurance carriers are evaluating the economic consequences of reimbursement for e-mail consultation. E-mail consultation raises potential medico-legal concerns, including establishment of the physician-patient relationship, malpractice liability, and HIPAA compliance. CONCLUSIONS: The increasing prevalence of e-mail consultation raises new concerns for neurosurgeons. Some of these concerns have yet to be addressed by regulatory commissions or in the courts. If used appropriately, e-mail communication can facilitate physician-patient interactions, improve access to care, save time for each interaction, and possibly reduce costs of care. PMID- 15120223 TI - Lower dose intraventricular T-PA fibrinolysis: case report. AB - BACKGROUND: Recombinant tissue plasminogen activator (rtPA) intraventricular fibrinolysis has been demonstrated to be efficacious in clearing blood from the ventricular system. Preliminary studies indicate it may improve survival. There have also been reports of adverse affects from intraventricular fibrinolysis. Optimal dosing of rtPA has not been established. METHODS: A 40-year-old patient with intraventricular hemorrhage extension secondary to a ruptured aneurysm was treated with a one-time infusion of 1 mg of rtPA through a right ventriculostomy. RESULTS: Computed tomography scans demonstrated excellent resolution of intraventricular blood and improvement in cerebral spinal fluid flow after fibrinolysis. Fibrinolysis was most marked in the third and fourth ventricles. There were no adverse effects noted. CONCLUSIONS: Intraventricular fibrinolysis is effective at a lower dose than previously used. Lower doses may have fewer adverse affects. PMID- 15120225 TI - Cranial-epidural tuberculosis presenting as a scalp swelling. AB - BACKGROUND: Unlike the brain tuberculoma, tubercular osteomyelitis of the skull is very rare and not sufficiently described in the literature. Awareness of this entity makes diagnosis possible. CASE DESCRIPTIONS: Two unique cases of cranial and epidural tuberculosis (TB) with absence of intradural and brain involvement are presented. Both patients presented with scalp swellings but extending through the calvarium into the epidural space. Histologic/bacteriologic confirmation of tuberculosis was obtained from biopsy specimens. Magnetic resonance imaging (MRI) findings of this rare lesion are described for the first time. CONCLUSIONS: Inflammatory scalp lesions with skull involvement and epidural extension should be investigated for tuberculous etiology. With early diagnosis and a combination of surgical and medical management, all cases of skull tuberculosis are potentially curable. PMID- 15120227 TI - Symmetric dumbbell ganglioneuromas of bilateral C2 and C3 roots with intradural extension associated with von Recklinghausen's disease: case report. AB - BACKGROUND: Ganglioneuromas are rare benign tumors arising most commonly from the sympathetic nervous system. They occasionally grow in a dumbbell fashion extending into the spinal canal extradurally. However, ganglioneuromas of the cervical spine with intradural extension or multiple locations or in association with von Recklinghausen's disease are rare. CASE DESCRIPTION: A 35-year-old man with von Recklinghausen's disease presented with tetraparesis and respiratory dysfunction. Preoperative neuroimaging revealed an intradural mass extending from the foramen magnum to the C4 vertebral level, as well as bilateral extravertebral extension connecting it with bilateral paraspinal lesions in a dumbbell fashion. Four intradural tumors associated with the bilateral C2 and C3 nerves and located ventrally were removed, leaving the intraforaminal and extradural portion intact. The procedure resulted in postoperative symptomatic improvement. Second, extravertebral tumors of the left neck, which were not related to the cervical sympathetic nerve, were removed. The pathologic diagnosis of the tumors of both the intradural space and cervical neck was ganglioneuroma. CONCLUSION: We present an extremely rare case in an adult with von Recklinghausen's disease who had bilateral, symmetric and multiple dumbbell ganglioneuromas with intradural extension, and also multiple bilateral ganglioneuromas at the neck. The intradural ganglioneuromas were suspected to have originated from the posterior root ganglions of the bilateral C2 and C3 nerves and to have extended ventrally to the spinal cord involving not only sensory but also motor rootlets; the ganglioneuroma of the neck was suspected to have originated from the cervical nerve itself. PMID- 15120229 TI - Tectal plate cavernoma-a special entity of brainstem cavernomas: case report. AB - BACKGROUND: Brainstem cavernous malformations (BCM) have a high incidence of bleeding and rebleeding and carry a high rate of neurologic morbility. Locations in the tectal plate that represent a small percentage of BCMs have rarely been reported in the literature. The authors present a case of a patient with such localization who was successfully operated. CASE DESCRIPTION: A 24-year-old male known for having a tectal plate cavernoma with obstructive hydrocephalus, previously treated by shunting in another hospital, was admitted in our institute because of increasing headaches, gradual drowsiness, and the inability to stand up. Investigations revealed a compressive cavernoma lateralized on the left side of the tectal plate and a residual hydrocephalus in spite of the previous shunting. A new shunting procedure did not improve clinical conditions. Thus, an aggressive surgical resection was decided upon and was performed through an occipital-transtentorial approach with the aid of intraoperative brainstem and middle latency auditory evoked potentials (BAEPs/MLAEPs) monitoring. Total resection was achieved without significant deterioration except a hypovoltage of wave V after stimulation of the right ear, demonstrating a left collicular dysfunction. The patient was discharged on the 36th day after surgery. Seven months later, audiometry was normal, in spite of the persistence of the hypovoltage of the V wave after stimulation of the right ear, and functional status appraised using the Karnofsky score was at 100%. Professional activity could be resumed. CONCLUSION: Tectal plate cavernomas (TPC) represent a special entity of BCM. They are surgically accessible lesions on the dorsal aspect of the brainstem. Our preferred approach is the occipital-transtentorial approach. The use of intraoperative auditory evoked potentials monitoring make the surgical resection safer. PMID- 15120231 TI - Transient cortical abnormalities on magnetic resonance imaging after status epilepticus: case report. AB - INTRODUCTION: Transient neuroimaging findings associated with seizure activity have received relatively little attention in the neurosurgical literature. These abnormalities may mimick neoplastic or ischemic changes on magnetic resonance imaging (MRI), possibly leading to additional studies and surgical treatment. CASE DESCRIPTION: A 17-year-old right-handed male was transferred to emergency room in status epilepticus. A MRI obtained 5 months before admission was negative. On the day of admission, he had multiple intermittent upper-left extremity partial seizures and prolonged secondary generalized seizures. An electroencephalogram (EEG) showed frequent epileptiform discharges over the right hemisphere posteriorly. A MRI study performed 2 days after admission revealed non hemorrhagic abnormalities involving the right occipital region that were hyperintense on fluid-attenuated inversion recovery (FLAIR) and T2 weighted sequences. The apparent diffusion coefficient map was unremarkable. Follow-up MRIs, 3 and 11 months after admission, showed complete resolution of these lesions. CONCLUSION: Imaging findings after status epilepticus may raise suspicion of ischemic or neoplastic lesions. These findings may be reversible. Further follow-up imaging may prevent unnecessary intervention. PMID- 15120233 TI - Double-endoscopic approach for management of convexity arachnoid cyst: case report. AB - BACKGROUND: Controversy exists about the best treatment modality for arachnoid cysts. Widely accepted neurosurgical options include craniotomy with open resection of the cyst walls, shunting procedures and stereotactic fenestration of the cyst cavity. Recently, neuroendoscopic approach showed its effectiveness for treatment of these lesions. CASE DESCRIPTION: In the presented case the large convexity arachnoid cyst was diagnosed in a 22-year-old woman with head dullness and local bone bulging. No neurologic signs were found. Surgery was performed by double-neuroendoscopic approach with simultaneous use of two flexible ventriculofiberscopes, which permitted wide resection of the inner cyst wall. Complete resolution of symptoms was noted soon after surgery. CONCLUSION: Double endoscopic approach might be useful for complex neuroendoscopic procedures that need different simultaneous surgical actions, particularly for cases of large arachnoid cysts. PMID- 15120237 TI - The Medpor sheet as a sellar buttress after endonasal transsphenoidal surgery: technical note. AB - BACKGROUND: A technique to use the Medpor (porous high-density polyethylene) sheet as an optimal implant to reconstruct the sellar floor or the sphenoid rostrum in endonasal transsphenoidal surgery is described. METHOD: After endonasal transsphenoidal surgery, a 0.4-mm thick Medpor sheet is cut with scissors to a round or elliptical piece, which is placed in the sellar epidural space to reconstruct the sellar floor and buttress the pituitary gland and the sellar packing. In cases of a complete absence of the sellar floor, the sphenoid sinus is packed with autologous fat and fibrin glue, which are buttressed at the defect of the sphenoid rostrum with the Medpor sheet. RESULTS: Ten patients underwent the reconstruction using the Medpor sheet in endonasal transsphenoidal surgery, and were then evaluated clinically and radiologically with magnetic resonance images (MR) postoperatively. The Medpor sheet was easy to cut with scissors and flexible to introduce through a small nostril. It was also sufficiently stiff to buttress the packing material in the sellar cavity or the sphenoid sinus. On MRI, the low-signal intensity of the implant was so remarkable without artifact helping to understand the postoperative image around the reconstructed area. There was no infection or granulomatous tissue reaction related to the implant. CONCLUSION: The Medpor sheet can be an optimal implant to reconstruct the sellar floor or the sphenoid rostrum after endonasal transsphenoidal surgery. PMID- 15120240 TI - The power of the doctor, the vulnerability of the patient, and informed consent. PMID- 15120242 TI - The keys to the future of healthcare everywhere. PMID- 15120241 TI - Hua Tuo, patron of surgeons, or how the surgeon lost his head! PMID- 15120243 TI - Informed consent for the neurosurgeon. PMID- 15120245 TI - Molecular and functional imaging of cancer: advances in MRI and MRS. PMID- 15120246 TI - A modified transorbital baboon model of reperfused stroke. PMID- 15120247 TI - MRI investigations of graft rejection following organ transplantation using rodent models. PMID- 15120249 TI - Cardiovascular magnetic resonance imaging: current applications and future directions. PMID- 15120250 TI - MRI of animal models of brain disease. PMID- 15120248 TI - Structural and functional optical imaging of angiogenesis in animal models. PMID- 15120251 TI - Magnetic resonance imaging in animal models of pathologies. PMID- 15120252 TI - Application of combined magnetic resonance imaging and histopathologic and functional studies for evaluation of aminoguanidine following traumatic brain injury in rats. PMID- 15120253 TI - Using 99mTc-sestamibi to evaluate the effects of a chemosensitizer on P glycoprotein in multidrug-resistant carcinoma cells. PMID- 15120254 TI - Vascular-targeted nanoparticles for molecular imaging and therapy. PMID- 15120255 TI - Production and applications of copper-64 radiopharmaceuticals. PMID- 15120256 TI - Generation of DOTA-conjugated antibody fragments for radioimmunoimaging. PMID- 15120257 TI - Preparation of magnetically labeled cells for cell tracking by magnetic resonance imaging. PMID- 15120258 TI - The application of magnetic resonance imaging and spectroscopy to gene therapy. PMID- 15120259 TI - Voxelation methods for genome scale imaging of brain gene expression. PMID- 15120260 TI - MR-intracranial compliance and pressure: a method for noninvasive measurement of important neurophysiologic parameters. PMID- 15120261 TI - Near-infrared spectroscopy and imaging of tumor vascular oxygenation. PMID- 15120262 TI - Measuring changes in tumor oxygenation. PMID- 15120264 TI - Mapping and taphonomic analysis of the Homo erectus loci at Locality 1 Zhoukoudian, China. AB - From a detailed analysis of published and unpublished sources, we constructed a digitized three-dimensional, stratigraphically-controlled excavation grid of Zhoukoudian Locality 1 in order to assess the spatial relationships of the excavated materials. All 15 fossil Homo erectus loci were mapped on the grid. Meter cubes were used in excavation starting in 1934, and Loci H through O, established between 1934 and 1937, were mapped to within 1 m(3)vertical and horizontal provenience. Loci A through G, established between 1921 and 1933, were excavated in the northernmost part of Locality 1 by unmapped quarrying, but their stratigraphic levels were recorded. We could localize Loci A through G on the grid system by utilizing locations of remaining walls, stratigraphic sections, excavation reports, excavation maps, and photographs. Loci contained skeletal elements of Homo erectus individuals scattered over areas of the cave floor of up to 9 m in diameter. Scoring of taphonomic damage on the Homo erectus sample, as observed on casts and originals, demonstrates that 67% of the hominid sample shows bite marks or other modifications ascribed to large mammalian carnivores, particularly the large Pleistocene cave hyena, Pachycrocuta brevirostris. Virtually all of the remaining Homo erectus skeletal assemblage shows breakage consistent with this taphonomic pattern of fragmentation. Bioturbation by digging carnivores is the most likely explanation for a fragment of Homo erectus Skull XI discovered 1 m below its other conjoined portions in Locus L. Carbon on all the Homo erectus fossils from Locus G, a circumscribed area of 1-meter diameter, earlier taken to indicate burning, cooking, and cannibalism, is here interpreted as detrital carbon deposited under water, perhaps the result of hyaenid caching behavior. Locus G records the close stratigraphic and horizontal association of stone artifacts with Homo erectus and other vertebrate skeletal elements, an association that is seen at other loci as well. Layer 4 of the excavation contains equid cranial bone previously interpreted to have been burned while fresh. We here document that Locus B Homo erectus, including Skull I, is stratigraphically associated with this evidence, but at some 10-12 m distance. Even though the presence of wood-stoked fires and hearths is not supported by geochemical results, evidence of fire at Locality 1 in the form of burned bone is confirmed. Contextual relationships of fossil skeletal elements, relationships of carnivore damage and stone tool cutmarks on bone, and evidence of the burning of fresh bone associated with Homo erectus and stone tools support a model of transient hominid scavenging aided by the use of fire at the large hyenid den that became Zhoukoudian Locality 1. Although the original excavation catalogue from Locality 1, as well as a significant number of fossils and stone artifacts, were lost during World War II, catalogue numbers on the many surviving specimens can be used to locate fossils and artifacts within the three-dimensional grid provided in this paper. PMID- 15120265 TI - An examination of dental development in Graecopithecus freybergi (=Ouranopithecus macedoniensis). AB - This study examined enamel thickness and dental development in Graecopithecus freybergi (=Ouranopithecus macedoniensis), a late Miocene hominoid from Greece. Comparative emphasis was placed on Proconsul, Afropithecus, Dryopithecus, Lufengpithecus, and Gigantopithecus, fossil apes that vary in enamel thickness and patterns of development. In addition, comparisons were made with Paranthropus to investigate reported similarities in enamel thickness. Several sections of a right lower third molar were generated, from which enamel thickness and aspects of the enamel and dentine microstructure were determined. Data from parallel sections shed light on the effects of section obliquity, which may influence determination of both enamel thickness and crown formation time. Graecopithecus has relatively thick enamel, greater than any fossil ape but less than Paranthropus, with which it does show similarity in prism path and Hunter Schreger band morphology. Aspects of enamel microstructure, including the periodicity and daily secretion rate, are similar to most extant and fossil apes, especially Afropithecus. Total crown formation time was estimated to be 3.5 years, which is greater than published values for modern Homo, similar to Pan, and less than Gigantopithecus. Data on dentine secretion and extension rates suggest that coronal dentine formation was relatively slow, but comparative data are very limited. Graecopithecus shares a crown formation pattern with several thick-enamelled hominoids, in which cuspal enamel makes up a very large portion of crown area, is formed by a large cell cohort, and is formed in less than half of the total time of formation. In Paranthropus, this pattern appears to be even more extreme, which may result in thicker enamel formed in an even shorter time. Developmental similarities between Paranthropus and Graecopithecus are interpreted to be parallelisms due to similarities in the mechanical demands of their diets. PMID- 15120266 TI - Protostylid variation in Australopithecus. AB - Recent advances in computed tomography (CT) and genetics provide new insights into the morphology and biology of anatomical traits, particularly in the dentition. As we move towards a fuller understanding of the genetic and developmental bases for dental traits, we need to reassess the taxonomic and evolutionary variation of established characters. Quantitative genetic analyses indicate that the degree of expression of upper and lower primate cingular remnants are genetically interdependent. This has serious evolutionary implications that need to be explored for fossil hominids. Studies of Carabelli's cusp, a cingular remnant on hominid upper molars, have been advanced through both genetic and CT analyses setting the stage for such an investigation. But its mandibular morphological homologue, the protostylid has not been similarly studied. This paper represents the first step towards a quantitative understanding of the variation and evolution of this trait in early hominids. Since the first discoveries of Australopithecus specimens in South Africa more than sixty years ago, cingular features on lower molars have played a significant role in the description and comparison of hominid taxa. This largely qualitative history is reviewed. Because the modern human classification system for protostylid variation does not adequately describe the variation seen in Australopithecus samples, a quantification scheme with six expression states is established. Using this new protocol, protostylid variation in six species of Australopithecus is assessed. Results from these analyses show that the distribution of the degree of protostylid expression in these species is highly varied. When first, second, and third molar samples are considered separately, the distribution of expression states is found to differ considerably within the same species. These results provide a foundation for further genetic and developmental research on the evolutionary history of the hominid dentition. PMID- 15120267 TI - Beyond leopards: tooth marks and the contribution of multiple carnivore taxa to the accumulation of the Swartkrans Member 3 fossil assemblage. AB - The ca. 1.0 myr old fauna from Swartkrans Member 3 (South Africa) preserves abundant indication of carnivore activity in the form of tooth marks (including pits) on many bone surfaces. This direct paleontological evidence is used to test a recent suggestion that leopards, regardless of prey body size, may have been almost solely responsible for the accumulation of the majority of bones in multiple deposits (including Swartkrans Member 3) from various Sterkfontein Valley cave sites. Our results falsify that hypothesis and corroborate an earlier hypothesis that, while the carcasses of smaller animals may have been deposited in Swartkrans by leopards, other kinds of carnivores (and hominids) were mostly responsible for the deposition of large animal remains. These results demonstrate the importance of choosing appropriate classes of actualistic data for constructing taphonomic inferences of assemblage formation. In addition, they stress that an all-encompassing model of assemblage formation for the hominid bearing deposits of the Sterkfontein Valley is inadequate and that each must be evaluated individually using not just analogical reasoning but also incorporating empirical data generated in the preserved fossil samples. PMID- 15120268 TI - Dental topography and diets of Australopithecus afarensis and early Homo. AB - Diet is key to understanding the paleoecology of early hominins. We know little about the diets of these fossil taxa, however, in part because of a limited fossil record, and in part because of limitations in methods available to infer their feeding adaptations. This paper applies a new method, dental topographic analysis, to the inference of diet from fossil hominin teeth. This approach uses laser scanning to generate digital 3D models of teeth and geographic information systems software to measure surface attributes, such as slope and occlusal relief. Because it does not rely on specific landmarks that change with wear, dental topographic analysis allows measurement and comparison of variably worn teeth, greatly increasing sample sizes compared with techniques that require unworn teeth. This study involved comparison of occlusal slope and relief of the lower second molars of Australopithecus afarensis (n=15) and early Homo (n=8) with those of Gorilla gorilla gorilla (n=47) and Pan troglodytes troglodytes (n=54). Results indicate that while all groups show reduced slope and relief in progressively more worn specimens, there are consistent differences at given wear stages among the taxa. Early Homo shows steeper slopes and more relief than chimpanzees, whereas A. afarensis shows less slope and relief than any of the other groups. The differences between the two hominin taxa are on the same order as those between the extant apes, suggesting similar degrees of difference in diet. Because these chimpanzees and gorillas differ mostly in fallback foods where they are sympatric, results suggest that the early hominins may likewise have differed mostly in fallback foods, with A. afarensis emphasizing harder, more brittle foods, and early Homo relying on tougher, more elastic foods. PMID- 15120269 TI - Tooth loss, survival, and resource use in wild ring-tailed lemurs (Lemur catta): implications for inferring conspecific care in fossil hominids. PMID- 15120270 TI - 2D or not 2D, and other interesting questions about enamel: reply to Macho et al. (2003). PMID- 15120271 TI - Adsorption of oxygen by heat-treated granular and fibrous activated carbons. AB - Three granular and one fibrous activated carbon with different physicochemical characteristics were subjected to heat treatment at 900 degrees C under vacuum or hydrogen flow. Subsequently, oxygen chemi- and physisorption isotherms were volumetrically obtained at 34 degrees C. Oxygen sorption experiments showed lower amounts of oxygen uptake by the H(2)-treated than by the vacuum-treated carbons, indicating that H(2) treatment effectively stabilized the surfaces of various carbons tested in this study. At low pressures, from approximately 0.001 to approximately 5 mmHg, adsorption of oxygen was governed by irreversible chemisorption, which was well described by the Langmuir equation. At higher pressures oxygen uptake occurred as a result of physisorption, which was in agreement with Henry's law. Kinetic studies showed that oxygen chemisorption was affected by both carbon surface chemistry and porosity. The results indicated that oxygen chemisorption initially started in the mesopore region from the high energetic sites without any mass transfer limitation; thus a constant oxygen uptake rate was observed. Once the majority of these sites were utilized, chemisorption proceeded toward the less energetic sites in mesopores as well as all the sites located in micropores. As a result, an exponential decrease in the oxygen uptake rate was observed. PMID- 15120272 TI - Formation and stability of rare earth metalloporphyrin monolayer films on amino terminated silanized quartz surfaces studied by ultraviolet-visible and X-ray photoelectron spectroscopy. AB - We investigated the fabrication of self-assembled monolayers of 5,10,15,20-tetra (p-chlorophenyl)-porphyrin metal hydroxyl compounds (MOH; M=Gd, Tb, Er, Lu) on amino-terminated silanized quartz surfaces using ultraviolet-visible (UV-vis) and X-ray photoelectron spectroscopy (XPS). The orientation of MOH molecules in the films, the kinetics of the adsorption of MOH from a chloroform solution on an amino-terminated quartz substrate, and the stability of the metalloporphyrin films under natural light, UV light, and acidic and basic conditions were studied by UV-vis spectroscopy. The results indicate that the central metal is crucial in the formation of self-assembled porphyrin films and that the stability of the MOH SAMs also depends on the central metal. Under natural and UV light irradiation conditions, the stability of the MOH SAMs depends on the strength of the M-N bonds between the central metal and pyrrole nitrogens or between the central metal and the axial ligand. In the acidic conditions, the UV-vis spectra of the MOH SAMs show decreases in the absorbance and blue shifts. These spectral changes lead us to suggest that the four M-N bonds between the central metal and the pyrrole nitrogens of the porphyrins are cleaved and the resulted porphyrins are protonated under acidic conditions. Thus, it is likely that the protonated porphyrins and MOH form pi-pi complexes with a parallel stacking of the macrocycles through the electrostatic attractive interaction in the SAMs, resulting in the blue shifts of the Soret bands of the SAMs. PMID- 15120273 TI - Passivation of the calcite surface with malonate ion. AB - Samples of polycrystalline calcite were impregnated with solutions of malonic acid of three concentrations (5 x 10(-2), 5 x 10(-3) , and 5 x 10(-4) M) and different pH values (6.00, 7.00, and 8.00). The impregnation was carried out at room temperature to evaluate the adsorption of malonate ion in the calcite surface to optimize the conditions for possible application on limestone and marble in cultural heritage materials. The affinity of the malonate ion was determined through the potentiometric measurement of the surface charge and the corresponding adsorbed amounts by titration, Raman spectroscopy, and small-angle X-ray scattering (SAXS). The results indicate effective adsorption of the malonate ion on the surface at a pH value close to the point of zero charge (pHpzc approximately 8.20) and changes in some surface morphological properties such as the pore shape and the pore size distribution. The presence of a malonate adsorptive layer on calcite generates an interface interaction potential that may influence the reaction and transport mechanisms within the medium. PMID- 15120274 TI - H2O outgassing from silica-filled polysiloxane TR55. AB - Temperature-programmed desorption/decomposition (TPD) was employed to obtain the moisture content and outgassing kinetics of TR55, a silica-filled cross-linked polysiloxane. The total moisture content of TR55 in the as-received state and after 20-30 min of vacuum pumping in the load-lock prior to TPD was measured to be on the order of 0.35 wt%. Physisorbed H(2)O and chemisorbed H(2)O account for about 13.2 and 86.8%, respectively, of the 0.35 wt% measured moisture content. H(2)O outgassing models based on the kinetics measured from TPD experiments suggest that loosely bound chemisorbed water outgasses in a dry environment slowly but continuously over many decades at or a little above room temperature. However, physisorbed water can be easily pumped out in a matter of hours at around 400 K. PMID- 15120275 TI - Effects of chemical functional groups on the polymer adsorption behavior onto titania pigment particles. AB - The effects of functional groups on polymer adsorption onto titania pigment particles have been investigated as a function of pH and ionic strength using polyacrylic acid and modified polyacrylamides. The polyacrylamides include the homopolymer, an anionic copolymer with hydroxyl and carboxylate group substitution, and a nonionic copolymer with hydroxyl group substitution. Adsorption isotherms and infrared spectroscopy were used to examine the polymer pigment interactions. The adsorption of the polyacrylic acid and anionic polyacrylamide on titania pigment is greatest when electrostatic repulsion is absent or reduced. At low pH values, below the pigment isoelectric point (IEP), or at high ionic strength, the adsorption density of the anionic polymers on titania pigment is high, while at higher pH values above the pigment IEP, the adsorption density decreases. But the adsorption of nonionic polymers on titania pigment is not influenced by either ionic strength or pH. Acrylamide groups were found to hydrogen bond with the titania pigment surface, independent of pH. With the inclusion of hydroxyl functional groups into the polyacrylamide chain, the polymer adsorption density increased without increased adsorption affinity. Carboxylate functional groups in the anionic polymers strongly interact with the pigment surface, producing the highest adsorption density at low pH values. All polymers exhibit Langmuir adsorption behavior with hydrogen bonding found as the dominant mechanism of adsorption in addition to electrostatic interaction occurring for the anionic polymers. PMID- 15120277 TI - Swelling behavior of PMMA-g-PEO microgel particles by organic solvents. AB - Aqueous dispersions of cross-linked poly(methylmethacrylate)-g-poly(ethylene oxide) [PMMA-g-PEO] microgel particles have been prepared from mixtures of methylmethacrylate [MMA] and MMA-PEO macromonomer, with ethylene glycol dimethacrylate [EGDM] as the cross-linking monomer (0.2-0.5% wt%). The hydrodynamic radius of these (unswollen) microgel particles ranged from 73 to 85 nm, and the particles were essentially monodisperse with regard to their size distribution. Their swelling behavior has been investigated in the presence of both water-miscible and water-immiscible organic solvents. In general, with the addition of a water-miscible solvent, deswelling behavior was observed. However, the microgel particles were swollen on addition of 1,4-dioxan, which is a good solvent for PMMA. With water-immiscible organic solvents, the extent of swelling depended on the solvency properties of the organic liquid for PMMA. In the presence of benzene, the somewhat large increases in particle size have been attributed to weak flocculation. This has been assumed from an estimate of the van der Waals attraction energy between the swollen microgel particles. PMID- 15120276 TI - Synthesis and characterization of tri(ethylene oxide)-attached poly(amidoamine) dendrimer layers on gold. AB - This paper describes the synthesis of a tri(ethylene oxide)-attached fourth generation poly(amidoamine) dendrimer (EO3-dendrimer) and the characterization of its layers on gold. NMR analysis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry revealed that about 61 amine groups of a G4 PAMAM dendrimer were covalently conjugated with tri(ethylene oxide) units, accounting for a 95% modification level. Layers of the EO3-dendrimer were formed on gold, and the resulting surface was characterized by infrared reflection absorption spectroscopy, ellipsometry, and contact angle goniometry. The EO3 dendrimer resulted in more hydrophilic and less compact layers with no substantial deformation of the molecule during layer formation by virtue of the EO3 units, compared to a PAMAM dendrimer. Interestingly, the specific binding of avidin to the biotinylated layers of the EO3-dendrimer approached a surface density of 5.2 +/- 0.2 ngmm-2, showing about 92% of full surface coverage. The layers of the EO3-dendrimer were found to be more resistant to nonspecific adsorption of proteins than PAMAM dendrimer layers when bovine serum albumin and serum proteins were tested. PMID- 15120278 TI - Effects of lipid chain length on molecular interactions between paclitaxel and phospholipid within model biomembranes. AB - Molecular interactions between an anticancer drug, paclitaxel, and phosphatidylcholine (PC) of various chain lengths were investigated in the present work by the Langmuir film balance technique and differential scanning calorimetry (DSC). Both the lipid monolayer at the air-water interface and lipid bilayer vesicles (liposomes) were employed as model biological cell membranes. Measurement and analysis of the surface pressure versus molecular area curves of the mixed monolayers of phospholipids and paclitaxel under various molar ratio showed that phospholipids and paclitaxel formed a nonideal miscible system at the interface. Paclitaxel exerted an area-condensing effect on the lipid monolayer at small molecular surface areas and an area-expanding effect at large molecular areas, which could be explained by the intermolecular forces and geometric accommodation between the two components. Paclitaxel and phospholipids could form thermodynamically stable monolayer systems: the stability increased with the chain length in the order DMPC (C14:0)>DPPC (C16:0)>DSPC (C18:0). Investigation of paclitaxel penetration into the pure lipid monolayer showed that DMPC had a higher ability to incorporate paclitaxel and the critical surface pressure for paclitaxel penetration also increased with the chain length in the order DMPC>DPPC>DSPC. A similar trend was testified by DSC studies on vesicles of the mixed paclitaxel/phospholipids bilayer. Paclitaxel showed the greatest interaction with DMPC while little interaction could be measured in the paclitaxel/DSPC liposomes. Paclitaxel caused broadening of the main phase transition without significant change at the peak melting temperature of the phospholipid bilayers, which demonstrated that paclitaxel was localized in the outer hydrophobic cooperative zone of the bilayer. The interaction between paclitaxel and phospholipid was nonspecific and the dominant factor in this interaction was the van der Waals force or hydrophobic force. As the result of the lower net van der Waals interaction between hydrocarbon chains for the shorter acyl chains, paclitaxel interacted more readily with phospholipids of shorter chain length, which also increased the bilayer intermolecular spacing. PMID- 15120279 TI - Immobilization of biogenic gold nanoparticles in thermally evaporated fatty acid and amine thin films. AB - We have recently demonstrated the biological synthesis of gold nanoparticles by the reduction of aqueous chloroaurate ions by the fungus Fusarium oxysporum and with extract of geranium (Pelargonium graveolens) leaf. In this paper, we demonstrate the immobilization of biogenic gold nanoparticles in lipid thin films deposited by thermal evaporation. The charge on the gold nanoparticles synthesized by both the fungus and the geranium plant extract is used to facilitate their immobilization in both anionic and cationic lipid thin films. A rough estimate of the isoelectric point of the proteins capping the gold nanoparticles synthesized using the fungus could be made by pH-dependent microgravimetry studies of the immobilization process. An interesting size and shape selectivity in the immobilized gold nanoparticles is observed in the lipid thin films. The biogenic gold nanoparticle-lipid composite films were characterized using quartz crystal microgravimetry, UV-vis absorption spectroscopy, Fourier transform infrared (FTIR) spectroscopy, and transmission electron microscopy. PMID- 15120280 TI - Wide-frequency-range dielectric response of polystyrene latex dispersions. AB - In this work we analyze the dielectric properties of dilute colloidal suspensions of nonconducting spherical particles with a thin electrical double layer from experimental data obtained by performing impedance spectroscopy experiments over a broad frequency range, from 20 Hz to 1 GHz. The electrode polarization correction was made by fitting a circuit model in the complex impedance plane (impedance spectrum) using a constant phase angle (CPA) element to fit the electrode polarization in series with the sample impedance. This simple procedure is found to be effective in eliminating the electrode contribution. The dielectric response shows two different dispersions, the alpha relaxation (counterion relaxation) that occurs at low kilohertz frequencies, and the delta relaxation (Maxwell-Wagner effect) found in the MHz range. These are reasonably well fitted over a broad frequency range by the theoretical expressions given by a simplified standard model (not including anomalous conduction) and a generalized model (including anomalous conduction) for the low-frequency dispersion, plus Maxwell-Wagner-O'Konski theory for the delta relaxation in the mid-frequency range. An analysis was also made of the need to include, for these latices, the effects of ion mobility in the Stern layer in order for the values of the zeta-potential obtained from electrophoretic and dielectric data to be compatible with each other. PMID- 15120281 TI - Mechanism of growth of colloidal silver nanoparticles stabilized by polyvinyl pyrrolidone in gamma-irradiated silver nitrate solution. AB - Silver nanoparticles were prepared by using polyvinyl pyrrolidone (PVP) as a stabilizer and gamma-irradiation. Transmission electron microscopy (TEM) results showed that both the amount and the molecular weight of PVP in the irradiated solution considerably affect the average size of the silver nanoparticles. The average size of the silver nanoparticles decreases with increasing the amount of PVP in the solution, but increases with increasing its molecular weight. Further, TEM showed that the silver nanoparticles become disassembled into smaller nanoparticles after dilution with distilled water and sonication. Since the processes of dilution and sonication are not expected to result in chemical reactions or to split the silver nanoparticles, we conclude that each silver nanoparticle prepared by [Formula: see text] -irradiation consists of several smaller nanoparticles surrounded by PVP. Thus, based on these observations, we propose a three-step mechanism for the growth of the silver nanoparticles under the conditions considered here. In the first step, the silver ions interact with PVP, then in the second step the silver ions that are exposed to gamma irradiation are reduced to silver atoms; nearby silver atoms then aggregate at close range. These aggregates are the primary nanoparticles. Finally, these primary nanoparticles coalesce with other nearby primary nanoparticles or interact with PVP to form larger aggregates which are the secondary (final) nanoparticles. PMID- 15120282 TI - Preparation and physicochemical and electrochemical characterization of exfoliated graphite oxide. AB - Exfoliated graphite oxide (EGO) is prepared by oxidizing exfoliated graphite (EG) using a mixture of KMnO(4)/H(2)SO(4). The physicochemical characterization of the EGO has been carried out using FT-Raman, FT-IR, XPS, NMR, and diffraction techniques. Colloidal form of EGO is subsequently prepared by ultrasonicating EGO in water. Thin films of EGO on a glassy carbon/gold surface are formed and the electrochemical and ion exchange properties have been studied using various redox systems such as K(4)[Fe(CN)(6)], ascorbic acid, and dopamine. The charge-based adsorption properties can be made use of, to either suppress or catalyze ascorbic acid oxidation. Adsorption and preconcentration of dopamine on the EGO film has been shown to electrocatalyze the oxidation of NADH. PMID- 15120283 TI - Nonspherical ZnS colloidal building blocks for three-dimensional photonic crystals. AB - The asymmetry introduced by a complex or nonspherical basis promotes photonic band gap formation in three-dimensional photonic crystals. However, relatively few techniques have been demonstrated to produce uniform nonspherical colloids for use as photonic crystal bases. Here we expand the menu of basis types with high refractive index by preparing nonspherical zinc sulfide colloids of uniform size and shape. Dimers, trimers, and planar tetramers were precipitated from aqueous solution by the thermal decomposition of thioacetamide in the presence of zinc nitrate, manganese nitrate, and nitric acid. The well-defined morphological types were obtained from suspensions aged for 4-6 h at 26-32 degrees C and then for 20-35 min at 85 degrees C. Stereological techniques were used to analyze SEM images and determine the percentage of each particle class. For example, the quantitative characterization of a particle population prepared at 29 degrees C for 6 h and 85 degrees C for 22 min had the composition 59+/-3% spheres, 31+/-2% dimers, 7+/-1% trimers, 0.4+/-0.2% tetramers, and 2.5+/-0.8% complex clusters (encompasses all other varieties of shape). X-ray diffraction and X-ray photoelectron spectroscopy confirmed the zinc blend crystal structure and the stoichiometric composition of the particles. The refractive index was estimated as 2.25 (413 nm) -2.09 (709 nm) by fitting experimental absorption spectra to curves derived from Mie scattering calculations. This indicated an average porosity approximately 24%. Such colloids offer the potential to form diamond like lattices with large, stable photonic band gaps. PMID- 15120284 TI - The effect of ultrasound on the particle size and structural disorder of a well ordered kaolinite. AB - The present study examined the effect of sonication on the particle size and structure of a well-crystallized (KGa-1) kaolinite from Georgia. Sonication produced an important delamination effect as well as a reduction of the other particle-size dimensions. The experiments, carried out under different experimental conditions, showed that particle-size reduction can be controlled through different variables such as power of ultrasonic processor, amount of sample (kaolinite + water), and time of treatment. As a consequence of this particle-size reduction the surface area increases sharply with the sonication time from 8.5 to 83 m2/g after 20 h with the most energetic treatment. Contrary to what is observed in the grinding treatment, sonication did not cause the amorphization of kaolinite, as observed by XRD and FTIR data. Nevertheless, ultrasound treatment increased the structural disorder, which consisted in increases in the proportion of specific translations (-a/3+b/3) between adjacent layers in the first hours of treatment, followed by increases in the proportion of random translations between layers. PMID- 15120285 TI - Growth of BaWO4 fishbone-like nanostructures in w/o microemulsion. AB - BaWO(4) fishbone-like nanostructures with fourfold structural symmetry have been successfully grown in w/o microemulsion. The BaWO(4) fishbone-like nanostructures have four rows of nanorods, epitaxially grown on the stem and perpendicular to the stem. The obtained samples are characterized by means of XRD, TEM, HRTEM, and SEM. It is found that the water content has a large influence on the size of the product and the molar ratio between cations and anions plays an important role in the morphology of the product. It is assumed that site-selective surfactant adsorption may be responsible for the formation of the BaWO(4) fishbone-like nanostructures. PMID- 15120286 TI - Raman spectroscopy of p-hydroxybenzoic acid aqueous solution and surface unenhanced Raman scattering on silver colloid with ultraviolet excitation. AB - A high-quality Raman spectrum of p-hydroxybenzoic acid (PHBA) aqueous solution (10(-2) M) under ultraviolet (UV) excitation at 325 nm was obtained, which could not be observed with visible and near infrared (NIR) excitations due to the low concentration in aqueous solution. However, the surface-unenhanced Raman scattering of PHBA in silver colloid excited by ultraviolet was unexpectedly observed, which was quite different from the cases excited with NIR and visible light, by which the SERS effect was very remarkable. This indicated that the SERS of the PHBA-silver colloid system showed selectivity to excitation wavelength. The enhancement mechanisms at different excitation wavelength regions are discussed. PMID- 15120287 TI - Effect of the exchangeable cations on the spectral properties of methylene blue in clay dispersions. AB - Adsorption of a cationic dye, methylene blue (MB), on the surface of montmorillonite leads to the molecular aggregation of dye cations, reflected by significant changes of dye optical properties. Montmorillonite samples, saturated with various inorganic cations (mono-, bi-, and trivalent, including those of transition metals), were used. Influence of the exchangeable cations on the MB aggregation was tested. Various properties of cations were considered (charge, diameter, acidity, hydration energies). Both direct and potential indirect effects of the cations were taken into account, such as salting-out effect, influence of the ions on solvent polarity, influence on swelling, colloid properties of montmorillonite dispersions, cation hydration properties, hydrolysis, and interaction of the cations with the clay surface. The spectra of MB in dispersions of montmorillonite saturated with NH4+, K+, Rb+, and Cs+ were significantly different from those of other reaction systems. Direct association between large monovalent cations and basal oxygen atoms of silicate probably leads to a partial fixation of the cations, which affects the ion exchange reaction and dye aggregation. Thus, the presence of large monovalent cations leads to the formation of fewer ordered H-aggregates in favor of monomers and aggregates of lower size. In these cases, dye species absorbing light of low energies also appeared in significant amounts and were assigned to J-aggregates, characterized by a head-to-tail intermolecular association. PMID- 15120288 TI - Creation of active sites by impregnation of carbon fibers: application to the fixation of hydrogen sulfide. AB - Activated carbon fibers, which exhibit high specific area and numerous active surface sites, constitute very powerful adsorbents and are widely used in filtration to eliminate pollutants from liquid or gaseous effluents. The fibers studied in this work are devoted to the filtration of gaseous effluent containing very small amounts (few vpm) of hydrogen sulfide. Preliminary experiments evidenced that these fibers weakly adsorb hydrogen sulfide. To improve their fixation capacity toward H(2)S the activated fibers are impregnated in an aqueous solution of potassium hydroxide. The impregnation treatment usually takes place before activation but in this work it occurs at room temperature after activation of the fibers. A further thermal treatment is performed to increase the efficiency of the system. The overall treatment leads to the creation of basic sites showing a great activity for H(2)S gas in the presence of water vapor. The mechanism has been established by a series of characterizations before, during, and after the different operation units. The KOH deposited after impregnation is carbonated into KHCO(3) at room temperature and then decomposed into K(2)CO(3) during the thermal treatment. K(2)CO(3) and H(2)S dissolve in a liquid aqueous solution formed on the fiber surface. Then carbonate ions and H(2)S molecules react together almost completely to yield HS(-) species. As a consequence the sorption capacities of hydrogen sulfide on the impregnated fibers are much higher, even for small hydrogen sulfide volume fractions. PMID- 15120289 TI - Broad reactivity trends for oxygen-isotope exchange from the near-surface regions of some metal (hydr)oxide solids. AB - The flux of (18)O from suspensions of isotopically enriched Cr(III) and Rh(III) hydroxide solids at varying temperature and pH was measured in a series of experiments. Most of these solids are metal hydroxide nanospheres that have a large surface area and a narrow distribution in particle sizes and contain inert metals (Cr(III) and Rh(III)). Using rate data for dissolved multimeric complexes as a guide, the solids were enriched in (18)O under conditions that were intended to affect mostly bound water molecules (eta-OH(2)) at the surface, but this point could not be verified. Nevertheless, the fluxes of (18)O back into solution from the isotopically enriched surfaces indicate that increased pH, which partly deprotonates the surface, is surprisingly unimportant to the rate and does not measurably affect (18)O fluxes. Although these data are sparse, Rh(III) solids react at rates that are lower than for Cr(III) solids, and the rates of exchange for crystalline and amorphous solids are relatively close. The results indicate that rates of ligand exchange at these surface sites are controlled dominantly by the local metal-oxygen bond strengths and that long-range forces are relatively unimportant. These experiments also indicate a strategy for measuring rates of ligand exchange from solid surfaces. PMID- 15120290 TI - Simultaneous EQCM and diffuse reflectance UV-visible spectroelectrochemical measurements: poly(aniline-co-o-anthranilic acid) growth and property characterization. AB - Piezoelectric diffuse reflectance spectroelectrochemistry (PDRSEC), a new technique of diffuse reflectance spectroelectrochemistry (DRSEC) in combination with electrochemical quartz crystal microbalance (EQCM), was developed to study the electrochemical copolymerization of aniline and o-anthranilic acid in 1.0 mol l(-1) HClO4 and the properties of these copolymers. The DRSEC using an integral sphere was proven to possess a higher optical sensitivity at the unpolished piezoelectric quartz crystal electrodes used than the mirror reflectance spectroelectrochemistry mode. The copolymers grown from the copolymerization bath of different molar fractions of o-anthranilic acid (F1, relative to the total amount of the two monomers) showed intermediate properties between those of the homopolymers, which varied gradually with F1. The swelling/dissolution behavior of the copolymers vs solution pH was traced via the EQCM frequency and resistance signals, and its large dependence on F1 was found and discussed. In a HAc-NaAc buffer solution at pH 5.6, the amount of adsorbed lysozyme was found to be positively correlated with F1, via an EQCM impedance investigation, demonstrating the feasibility of using poly(aniline-co-o-anthranilic acid) as a load-adjustable immobilization matrix for cationic proteins. The novel PDRSEC method proposed is highly recommended for surface electrochemistry studies at relatively rough electrodes. PMID- 15120291 TI - Preparation, characterization, and catalytic activity of CoMo/gamma-Al2O3 catalysts prepared by equilibrium deposition filtration and conventional impregnation techniques. AB - A CoMo/gamma-Al(2)O(3) catalyst, prepared by depositing on the Al(2)O(3) carrier first the Mo species via equilibrium deposition filtration (EDF) and then the Co species by dry impregnation, was compared to three CoMo/gamma-Al(2)O(3) samples prepared using various conventional impregnation methods. All samples had the same composition, corresponding to an atomic ratio Co/(Co+Mo) equal to 0.3. The above samples were characterized using various physicochemical techniques (AAS, BET, DRS, LRS, XPS, TPR, and NO chemisorption), and their catalytic activity was determined using the hydrodesulfurization (HDS) of thiophene as a probe reaction. The EDF-prepared catalyst was about 30-43% more active in HDS than those prepared with the conventional impregnation techniques at all reaction temperatures studied. In contrast, the EDF catalyst exhibited the lowest hydrogenation activity. The higher HDS activity of the EDF sample is attributed to the higher number of active HDS sites formed on its surface. It is concluded that the increased number of active sites is due to the fact that the deposition of the Mo species by EDF results to a higher coverage of the support surface by supported molybdenum phase, which in turn, inhibits the formation of the catalytically inactive CoAl(2)O(4) and favors the dispersion of octahedral cobalt on its surface. PMID- 15120292 TI - Modeling of submicrometer aerosol penetration through sintered granular membrane filters. AB - We present a deep-bed aerosol filtration model that can be used to estimate the efficiency of sintered granular membrane filters in the region of the most penetrating particle size. In this region the capture of submicrometer aerosols, much smaller than the filter pore size, takes place mainly via Brownian diffusion and direct interception acting in synergy. By modeling the disordered sintered grain packing of such filters as a simple cubic lattice, and mapping the corresponding 3D connected pore volume onto a discrete cylindrical pore network, the efficiency of a granular filter can be estimated, using new analytical results for the efficiency of cylindrical pores. This model for aerosol penetration in sintered granular filters includes flow slip and the kinetics of particle capture by the pore surface. With a unique choice for two parameters, namely the structural tortuosity and effective kinetic coefficient of particle adsorption, this semiempirical model can account for the experimental efficiency of a new class of "high-efficiency particulate air" ceramic membrane filters as a function of particle size over a wide range of filter thickness and texture (pore size and porosity) and operating conditions (face velocity). PMID- 15120293 TI - Simultaneous thermal and surfactant-induced Marangoni effects in thin liquid films. AB - The deformation of a thin liquid film in the presence of a surfactant monolayer, varying temperature distributions, and limited mass flux is considered. Use of lubrication theory yields a coupled pair of partial differential equations for the film height and surfactant surface monolayer concentration. The long-wave stability of the isothermal film is examined over a wide range of parameter values. It is shown that droplet patterns are obtained under certain thermal conditions for both an isothermal and nonisothermal underlying substrate. For the case of a localized thermal gradient initially imposed at the air-liquid interface, severe film thinning beneath the heat source was observed, which was not accompanied by droplet formation; pseudo steady states are observed in this case. In all situations the surfactant is found to rigidify the air-liquid interface, retarding thermally driven flow, while evaporation (condensation) acts to destabilize (stabilize) the film. PMID- 15120294 TI - Surfactant-encapsulated polyoxometalloeuropate: polarized Eu3+ emission in the highly ordered self-organizing film. AB - An organic/inorganic composite, (DODA)(9)EuW(10)O(36).9H(2)O, can form a highly ordered self-organizing film with a layered structure and almost fully interdigitated alkyl chains at solid supports. EuW10O36(9-) is deduced to be oriented in its self-organizing film, resulting in its polarized emission, which offers an accessible approach to fabricating the functional thin films containing polyoxometalates. PMID- 15120295 TI - Modeling of separation of aqueous solutions of FeCl3 and AlCl3 by zeolite-clay composite membranes using a space-charge model. AB - In our earlier work, we reported the separation of FeCl3 from its aqueous solution and AlCl3 from its aqueous solution by analcime zeolite (Z1) membrane and its nitrated (Z2 membrane) and aminated (Z3 membrane) forms. Experimental data on the separation of aqueous solutions of FeCl3 and AlCl3 by zeolite-clay composite membranes has been simulated using the two-dimensional space-charge model. The computational requirement of the model has been considerably reduced by first obtaining a series solution of the nonlinear Poisson-Boltzmann equation. The effective pore radius of the membrane is taken as the one that gives the best fit to the experimental data, while the pore length is determined from the SEM photograph of the cross-sectional view of the membrane. The effective pore radii of the Z1, Z2, and Z3 membranes for FeCl3 solute are found to be 8.0, 7.0, and 5.0 nm, respectively, while for AlCl3 they are 4.5, 2.5, and 2.5 nm, respectively. These values are much less than the average of the pore size range values determined independently in an earlier work using the bubble point method and indicate partial blocking of the pores by these salts. The effective pore radius is larger for FeCl3 as compared to AlCl3 and decreases on modification. The intrinsic rejection is also found to decrease on modification. The permeate flux calculated from the model matches very well with the experimental values. PMID- 15120296 TI - Restricted diffusion in silica particles measured by pulsed field gradient NMR. AB - The restricted diffusion coefficient of water through porous silica is measured by pulsed field gradient (PFG) NMR as a function of loading in order to develop a model for self-diffusion at full pore filling in sol-gel-made porous silica particles. This model describes the pore or intraparticle diffusion coefficient as a function of particle porosity, tortuosity, and the steric hindrance applied on the molecules by the pore space. The particle morphology is characterized by nitrogen adsorption and an appropriate tortuosity model is chosen in comparison with literature data. To characterize the material, NMR relaxation and diffusion studies at different degrees of pore filling were carried out in relation to the silica/water adsorption isotherm. PMID- 15120297 TI - True molar surface energy and alignment of surface molecules. AB - By use of data for surface tension, for the first time a method is presented for calculating true molar surface properties of liquids: free energy, entropy, and enthalpy. These new data allow full comparison with other molar quantities, such as enthalpy and entropy of vaporization. All data are at the normal boiling point. There are differences in behavior between various classes of nonpolar compounds. Rare gases and tetrahydrides of Group 14 form a separate category. The results agree with the experimental findings that water and alcohol molecules are aligned with the -OH groups attached to the surface. The data indicate that hydrogen peroxide and 1,2-ethanediol also have one -OH group directed toward the surface and one directed out toward the vapor phase. Small straight-chain amines have some structure at the surface, but the larger ones behave like the corresponding alkanes. Very polar compounds, such as nitriles, nitro compounds, and aldehydes, have little or no increased degree of order of molecules at the surface. Except for hydrogen-bonded compounds, molecules at the surface have surroundings similar to those in the bulk liquid. PMID- 15120298 TI - Interaction of surfactants and aminoindophenol dye. AB - The interaction of dye and surfactants was studied by their spectroscopic and surface properties. Large bathochromic shift (15 nm) in the absorption spectrum was found for aminoindophenol dye at high pH in cationic surfactant, while there is no significant shift in anionic, zwitterionic and nonionic surfactant solutions. The static and dynamic surface properties show there is strong interaction in mixture of cationic surfactant and aminoindophenol dye. Interaction of dye and surfactants on surface and in solution is correlated to the intensity of dye deposition on fiber. The charge complex formation between cationic surfactant and aminoindophenolic dye delays the dye diffusion into keratin fiber. The stronger is the dye/surfactant interaction, the lower dye deposition and diffusion become. PMID- 15120299 TI - Fast emulsion droplet sizing using NMR self-diffusion measurements. AB - Emulsion droplet sizing using pulsed field gradient (PFG) nuclear magnetic resonance (NMR) is a well-established technique. Traditionally these measurements require total acquisition times of typically 5-20 min per sample, which severely limits our ability to use this method to study dynamic processes. Here we present the application and verification of an NMR pulse sequence, Difftrain, which enables emulsion droplet size distributions to be measured in 3-10 s. We have previously introduced applications of Difftrain (C. Buckley, K.G. Hollingsworth, A.J. Sederman, D.J. Holland, M.L. Johns, L.F. Gladden, J. Magn. Reson. 161 (2003) 112-117), including the droplet sizing of a single unimodal emulsion sample. In this paper, several model emulsions containing different oils are measured and the results compared directly with sizing provided by laser scattering. In this manner, the Difftrain method is verified and its possibilities and limitations are explored. Guidelines are proposed for the range of droplet sizes for which accurate results can be produced. The Difftrain technique opens up the possibility of studying non-equilibrium emulsions; a study of the in situ emulsification of a 21% v/v water-in-silicone oil emulsion is presented. PMID- 15120300 TI - Five-component food-grade microemulsions: structural characterization by SANS. AB - In this paper we present the structural characterization of a five-component food grade microemulsion containing Tween 80, R(+)-limonene, ethanol, glycerol, and water. Our main approach to investigating the microstructure of dense microemulsions, and how it can be influenced by the various components, was to employ small-angle neutron scattering and the new evaluation technique for dense, interacting systems, the Generalized Indirect Fourier Transformation. We started our investigation with the impact of glycerol and ethanol on Tween 80 micelles in water. We found that glycerol increases the aggregation number and withdraws the hydrating agents from the headgroup region of the surfactant, resulting in a higher packing density of molecules in a micelle at slightly increasing size. The same trend holds when the micelles are oil swollen and/or ethanol is present. Ethanol, on the other hand, redistributes mainly between water and the interface headgroup region of the surfactant. Part of it replaces surfactant molecules in the micelles, which increases the available interface and results in a higher number of micelles with shrinking size. The same trend holds when the micelles are oil swollen and/or glycerol is present in the aqueous phase. We also investigated samples along the dilution of a mixture of surfactant and oil phase (R(+)-limonene and ethanol), which can be diluted with aqueous phase (mixture of water and glycerol) without the occurrence of phase separation. In some samples of this dilution most probably bicontinuous structures are present. To elucidate this point, we also employed dynamic light scattering, viscosity, and conductivity measurements. PMID- 15120301 TI - Conductivity of water-in-oil microemulsions stabilized by mixed surfactants. AB - The electrical conductivity of D2O-in-n-heptane microemulsions stabilized by cationic/nonionic surfactant mixtures was studied as a function of D2O content, surfactant concentration, and surfactant mixture composition. The surfactants employed were cationic di-n-didodecyldimethylammonium bromide, DDAB, nonionic poly(oxyethylene) monododecyl ethers, C12EJ, with J=3-8 and 23, nonionic polymeric surfactants of the type PEO-PPO-PEO (Pluronic), and the reverse structure analogues (Pluronic R). Qualitative structural information was drawn from a comparison between the measured conductivity and that predicted by the charge fluctuation model for spherical droplets. The conductivity versus water content curves were found to be typical for water-in-oil systems composed of spherical droplets. From the effect of blending nonionic surfactant with DDAB on the measured conductivities, it was concluded that microemulsion conductivity is independent of the concentration of cationic surfactant (DDAB). This finding agrees well with theoretical microemulsion conductivity models. PMID- 15120302 TI - An experimental study on the rheological properties of aqueous ceria dispersions. AB - The rheological properties of aqueous ceria dispersions are studied experimentally. In particular, the effects of particle concentration, temperature, pH, and ionic strength are discussed. If the volume fraction is below 2%, ceria slurry exhibits Newtonian behavior, and for higher volume fractions, shear-thinning behavior is observed. The effect of temperature on the behavior of ceria slurry is found to be pH-dependent. If pHIEP, the viscosity slightly increases with increased temperature. A shift of IEP to a higher value of pH was observed for ionic strength, even for indifferent electrolytes. The influence of pH on the rheological properties of ceria slurry decreases if the ionic strength is high. The pH at which viscosity and yield stress are maximum coincide with IEP only for low ionic strengths. The slopes of acidic and basic branches of viscosity against pH and yield stress against pH curves are not symmetrical at high ionic strength, and the alkaline branch deviates significantly from Hunter's theory. PMID- 15120303 TI - Steady flow and viscoelastic properties of lubricating grease containing various thickener concentrations. AB - The flow and viscoelastic properties of a lubricating grease formed from a thickener composed of lithium hydroxystearate and a high-boiling-point mineral oil were investigated as a function of thickener concentration. The flow properties of grease were measured using continuous shear rheometry, while the viscoelastic properties were measured using oscillatory shear measurements. The flow properties show that grease is a shear-thinning fluid with a yield stress that increases with thickener concentration. At concentrations of lithium hydroxystearate greater than 5% by volume, the storage modulus, G', was found to be greater than the loss modulus, G", with both moduli increasing with increasing thickener concentration, below this critical concentration G" was greater than G'. Slip at the wall of the measuring platens was a major problem encountered during the rheological measurement of grease, this is hardly surprising, and greases are designed to slip in their lubricating functions. Therefore the measuring platens were roughened by sandblasting and significantly higher yield values were recorded with the roughened geometries. Creep experiments were also performed. In the creep test, yield stresses of greases could be obtained. Zero shear viscosity was also calculated from the creep experiment and as a result viscosities over nine orders of magnitude were obtained. The power law index of the scaling law of the elastic modulus and yield stress with increasing volume fraction was found to be 4.7+/-0.2 suggesting that the flocculation of the particles that compose the grease is likely to be of the chemically limited aggregation variety. PMID- 15120304 TI - Instability of the interface between thin fluid films subjected to electric fields. AB - The effect of an externally applied electric field on the stability of the interface between two thin leaky dielectric fluid films of thickness ratio and viscosity ratio ris analyzed using a linear stability analysis in the long-wave limit. A systematic asymptotic expansion is employed in this limit to derive the coupled nonlinear differential equations describing the evolution of the position of the interface between the fluids and the interfacial free charge distribution. The linearized stability of these equations is determined and the effect of the ratio of the conductivities, dielectric constants, thicknesses, and viscosities on the wavenumber of the fastest growing mode, kmax, and the growth rate of the most unstable mode, smax, is examined in detail. Specific configurations considered in previous studies, such as a perfect dielectric-air interface, leaky dielectric-air interface, etc., emerge as limiting cases from the general formulation developed in this paper. Our results show that the viscosity ratio, mur, does not have any significant effect on kmax for the interface between perfect and leaky dielectric fluids. In marked contrast, however, mur is shown to have a significant effect on the interface between two leaky dielectrics. Increasing mur from 0.1 to 10 could decrease kmax up to a factor of 5. In general, our results show that the presence of nonzero conductivity in either one or both of the fluids has a profound influence on the length-scale characteristic of the linear instability: a reduction even by a factor of 1/50 in the length scale can be effected when compared to the interface between two perfect dielectrics. These predictions could have important implications in pattern formation applications in thin fluid films that employ electric fields. The variation of kmax and smax on the thickness ratio, beta, indicates in general that kmaxalpha(beta-apha), and smaxalpha(beta-theta), where the exponents alpha and theta (both >0) are found to depend only on the ratio of conductivities, and are largely independent of other system parameters. PMID- 15120305 TI - Intrinsic charge and Donnan potentials of grafted polyelectrolyte layers determined by surface conductivity data. AB - In order to characterize grafted polyelectrolyte layers based on electrokinetic measurements a theory of the surface conductivity Ksigma was developed, starting from the model of thick polyelectrolyte layers with uniform segment distribution and dissociable groups with an unknown pK value. According to this model the inner part of the polyelectrolyte layer adjacent to the substrate is considered to be isopotential while the potential decay occurs in a zone near the solution side of the layer. A simple equation for the Donnan potential psiD as a function of pH, pK, electrolyte concentration C0, and volume charge density rho was obtained. In the derived equation Ksigma is directly related to psiD while the other terms have less influence on the magnitude of Ksigma and can be accounted for in a second approximation using psiD as determined from the measured Ksigma. Evaluation of the suggested model indicates that Ksigma measurements provide an effective method to characterize polyelectrolyte layers by analyzing the dependence of psiD on pH and C0: The magnitude of Ksigma yields information about the surface charge at complete dissociation of the ionizable groups. The dependence of Ksigma on pH and C0 can be used for the determination of the pK value of the dissociating functions and the segment volume fraction of the polyelectrolyte can be estimated using the measured value of rho. PMID- 15120306 TI - Electroviscous effects on pressure-driven flow of dilute electrolyte solutions in small microchannels. AB - A fundamental understanding of the flow characteristics of electrolyte solutions in microchannels is critical to the design and control of microfluidic devices. Experimental studies have shown that the electroviscous effect is appreciable for a dilute solution in a small microchannel. However, the experimentally observed electroviscous effects cannot be predicted by the traditional theoretical model, which involves the use of the Boltzmann distribution for the ionic concentration field. It has been found that the Boltzmann distribution is not applicable to systems with dilute electrolyte solutions in small microchannels because it violates the ion number conservation condition. A new theoretical model is developed in this paper using the Nernst equation and the ion number conservation, instead of the Boltzmann distribution, to obtain the ionic concentration field. The ionic concentration field, electrical potential field, and flow field in small microchannels are studied using the model developed here. In order to verify this model, the model-predicted dP/dx (applied pressure gradient) Re (Reynolds number) relationship is compared with the experimentally determined dP/dx approximately Re relationship. Strong agreement between the model predictions and the experimental results supports this model. PMID- 15120307 TI - Theoretical prediction of surface tension of ternary liquid system (nitrogen + oxygen + argon) at elevated temperature and different pressure. AB - The surface tension of a ternary liquid mixture of nitrogen, oxygen, and argon has been calculated by a new approach using critical constants Pc, Vc, and Tc at temperatures ranging from 90 to 110 K and pressure up to 865 kPa. New formalism has been made by modifying the Brock-Bird relation. The computed values are compared with the experimental findings. Satisfactory results have been obtained. PMID- 15120308 TI - Brownian sampling in an unbounded space. AB - As a particle undergoes translational Brownian motion in an unbounded space, the particle samples the space. Traditionally the sampling in N dimensions is quantified in terms of average squared distance traversed (). However, another quantitative measure of the sampled space is the total number (n) of equispaced regions (of size LN) sampled after a particle moves with a diffusion coefficient (D) for a time (t). Calculations show that the average =a(Dt/L2)b. Results are given for a and b for 1, 2, 3, and 4 dimensions. PMID- 15120309 TI - Structural studies of Na-montmorillonite exchanged with Fe2+, Cr3+, and Ti4+ by N2 adsorption and EXAFS. AB - The structures of Fe(2+)-, Cr(3+)-, and Ti(4+)-modified montmorillonite prepared from ion exchange of the Na-clay with Fe(2+), Cr(3+), and Ti(4+) were investigated. Conventional BET surface area and spectroscopic analysis by extended adsorption fine structure (EXAFS) were applied. It was shown that the BET surface area of Na-clay was similar to that of Fe-clay, but somewhat different from those of Cr- and Ti-clay; it decreased in the order Na- > Fe- > Ti > Cr-montmorillonite. This sequence appeared to be consistent with the ion size Na(+) (0.95 nm)>Fe(2+) (0.65 nm)>Cr(3+) (0.62 nm), except for Ti(4+) (0.69 nm). EXAFS data showed that some Si atoms within montmorillonite were replaced by Ti atoms and that a neostructure of titanium oxide was formed. PMID- 15120310 TI - Proton magic-angle spinning-NMR investigation of surfactant aqueous suspensions. AB - In this Note we present the advantages of 1H magic-angle spinning nuclear magnetic resonance (MAS-NMR) for the investigation of surfactant suspensions via transverse relaxation rate (R2) measurements. 1H-relaxation rates can be determined by the classical CPMG method from high-resolution spectra obtained either under conditions of liquid-state NMR for monomers and small spherical micelles or by using MAS-NMR for larger aggregates. For a mixture of alkyl dioxyethylene sulfate and alkylbetaine (80:20, w/w), up to a percentage of surfactant in water of 20%, we found that R2 increased, in accordance with an increased micellar size and very likely the formation of an HI phase. However, above 25%, R2 decreased. This result suggests a change from a hexagonal to a lamellar phase that would be difficult to observe by proton NMR without magic angle spinning because the lines would be very broad, or by light scattering because of sample opacity. This NMR approach seems to have been overlooked by the community of surfactant physical chemists. It can be complementary to other analytical techniques and presents the advantage of not requiring isotopic labeling. PMID- 15120311 TI - Internal structure of clusters from charge heteroaggregation. AB - The internal structure of clusters formed by colloidal heteroaggregation of particles with opposite signs of charge is studied by means of computer simulations. Every particle is surrounded by a layer of particles of opposite sign, a second neighbors shell of particles mainly with the same sign, a third one of opposite sign, etc. As the distance from the particle increases, the system becomes more homogeneous and no difference between the numbers of particles with similar or opposite signs of charge can be noticed for distances larger than ten times the particle radius. For low ionic concentrations the local environment of particles is formed by quasi-straight branches, where the sign of charge alternates, and at high concentrations the structure of the cluster is typical of DLCA and the alternation is restricted to very short distances. However, this effect is not responsible for the low fractal dimensions observed in charge heteroaggregates. PMID- 15120312 TI - Hofmeister anion effect on aqueous phase behavior of heptaethylene glycol dodecyl ether. AB - The aqueous phase behavior of heptaethylene glycol dodecyl ether (C12E7) was investigated in the presence of sodium salts of Cl-, I-, and ClO4-. Pseudo binary T-X phase diagrams were constructed for these mixtures by means of differential scanning calorimetry. The salting-out electrolyte NaCl expanded the Lalpha region toward higher temperatures and shrank the H1 region toward lower temperatures compared with the salt-free system. On the contrary, the salting-in electrolytes NaI and NaClO4 induced shrinkage of the Lalpha region and an expansion of the H1 phase. The influence of these salts on the mesophase regions was more pronounced for the Lalpha phase than for the H1 phase, and area of the Lalpha phase region decreased in the sequence of NaCl > none > NaI > NaClO4, consist with the Hofmeister series of the anions. This salt effect on the mesophase stability in aqueous nonionic surfactant mixture would be qualitatively interpreted in terms of the salt effect on the hydration of the polyoxyethylene chain in the surfactant molecules. PMID- 15120313 TI - Modulation of cytokine production by some phthalimido-desmuramyl dipeptides and their cytotoxicity. AB - Muramyl dipeptide (MDP) is the smallest bacterial cell wall peptidoglycan component having immunomodulatory activity. In an attempt to obtain MDP derivatives with improved and better defined pharmacological profiles we synthesized a new lipophilic phthalimido-desmuramyl dipeptide, LK 508. This novel MDP analogue and three structurally related phthalimido-desmuramyl dipeptides (LK 413, LK 511 and LK 512) were evaluated immunologically. Their ability to modulate the production of cytokines was measured in vitro by their inclusion in cultures of human peripheral blood mononuclear cells (PBMC) activated by ionomycin and phorbol-12-myristate-13-acetate (PMA). The results were compared with the analogous activity of MDP. All compounds tested are strong up-regulators of IL-12 synthesis. All compounds except LK 512 also stimulated IFNgamma synthesis. LK 508, LK 511 and LK 512 are effective in up-regulating IL-2 production. LK 508 and LK 512 considerably up-regulate the synthesis of IL-4 and IL-10. LK 413 and MDP stimulated the production of Th1 promoting and Th1 (IFNgamma and IL-12) cytokines, while LK 508, LK 511 and LK 512 non-selectively up-regulated the production of both Th1 and Th2-types of (IL-4 and IL-10) cytokines. None of the phthalimido-desmuramyl dipeptides was cytotoxic in vitro against the normal cell line HUVEC (human endothelial cells) thereby indicating their potential for use in vivo. PMID- 15120314 TI - Synthesis and anticonvulsant activity of N-3 substituted 2,3-benzodiazepines. AB - A series of new 3-alkylcarbamoyl-1-aryl-3,5-dihydro-7,8-dimethoxy-4H-2,3 benzodiazepin-4-ones was synthesized starting from the corresponding 3-N unsubstituted derivatives, previously described as noncompetitive AMPA-type glutamate receptor antagonists. The new compounds proved to protect against seizures induced by means of auditory stimulation in DBA/2 mice and some of them showed anticonvulsant properties comparable or better than those of GYKI 52466, the prototype of 2,3-benzodiazepine noncompetitive AMPA receptor antagonists. PMID- 15120316 TI - Synthesis and cytotoxicity of a novel 1-alkylaminomethyl-2,4-diaryl-butadiene-1,3 fragment integrated within cyclohex(pent)enes. AB - A group of 1-alkylaminomethyl-2-aryl-3-arylidenecyclohex(pent)enes 3a-n with a 1 alkylaminomethyl-2,4-diaryl-1,3-butadiene fragment and a group of their congeners 3-alkylaminomethyl-1,2-diarylcyclohexene 7a-f have been synthesised for the first time. The conjugated system in 1-alkylaminomethyl-2-aryl-3 arylidenecyclohex(pent)enes 3a-n was unambiguously confirmed by X-ray crystallography. Cytotoxicity tests revealed that 3a-n possess inconsistent cytotoxicity against cancer cells, not their congeners 7a-f. PMID- 15120315 TI - A new class of NO-donor H3-antagonists. AB - Synthesis and pharmacological characterisation of a series of compounds obtained by joining, through appropriate spacers, NO-donor furoxan and nitrooxy moieties to the imidazole ring, as well as their structurally related analogues devoid of NO-donating properties are described. All the products were studied for their capacity to interact with H3-receptors present on the guinea-pig ileum and with H2-receptors present on guinea-pig right atrium. The whole series of products displayed reversible H3-antagonistic activity. No activity on H2-receptors was observed when the products were tested at 10 microM concentration. Many of the products were also able to induce partial relaxation when added to the bath after electrical contraction of the guinea-pig ileum during the study of their H3 antagonism. This phenomenon seems to be dependent on various factors; for some compounds it proved to be dependent on NO-mediated sGC activation, for other products it could be due to their weak M3-antagonism. The investigation of the lipophilic-hydrophilic balance of all the products indicates, for many of them, an ideal value to cross the blood-brain barrier. PMID- 15120317 TI - Design, synthesis and anticonvulsive activity of analogs of gamma-vinyl GABA. AB - For the development of new anticonvulsive agents, analogs of gamma-vinyl GABA (vigabatrin) containing GABA, gamma-vinyl GABA, valproic acid, nipecotic acid or isonipecotic acid moieties were prepared and evaluated for their anticonvulsive activities. Most of the prepared compounds showed moderate anticonvulsive activities. Among them compounds 10 and 16 displayed the most potent anticonvulsive activity and a broader spectrum compared to vigabatrin. PMID- 15120318 TI - Prediction of selectivity of alpha1-adrenergic antagonists by counterpropagation neural network (CP-ANN). AB - A quantitative structure-selectivity relationships of series of structurally diverse alpha1-adrenergic antagonists was performed by using counter-propagation neural network (CP-ANN). The theoretical molecular descriptors have been calculated and selected using CODESSA program. The results obtained for a highly non-congeneric set of molecules have confirmed the potential of use of CP-ANN approach in prediction of relative activity (selectivity) of alpha1-adrenergic antagonists. PMID- 15120319 TI - 1,5-Diarylsubstituted 1,2,3-triazoles as potassium channel activators. VI. AB - As part of our program toward designing potassium channel openers, synthesis of a novel series of 1,5-diphenylsubstituted 1,2,3-triazoles, as potential activators of the large-conductance calcium-activated potassium channels (BK), as well as their vasorelaxant activity are presented. The functional effect of these potential structurally novel BK-openers on vascular contractile function were studied in vitro, using isolated rat aortic rings pre-contracted with KCl 20 mM. Among the target compounds, only 16 showed appreciable effectiveness, exhibiting an efficacy parameter (57%) lower than that of NS1619 and a comparable potency index (pIC50: 5.58). PMID- 15120320 TI - Cytotoxicity of furoxans: quantitative structure-activity relationships study. AB - Furoxans are interesting biological active compounds. Recent studies demonstrate that they are cytotoxic under aerobic conditions. This paper shows that this cytotoxicity could be related with E(LUMO), calculated LogP and Mulliken charge on heterocycle nitrogen 2 calculated at ab initio level (3-21G* basis). It was possible to propose a tentative mechanism of cytotoxicity and structural modifications to modulate the cytotoxic potency of furoxan derivatives. PMID- 15120321 TI - Synthesis and in vitro antileukemic activity of new 4-triazenopyrazole derivatives. AB - Several new 4-(3,3-dimethyltriazeno)-5-benzamidopyrazole derivatives were prepared by reacting 4-diazo-5-benzamidopyrazole derivatives with dimethylamine. The compounds were tested at 10 microM for their vitro antileukemic activity against K562 (Human chronic myelogenous leukemia) and Raji (human Burkitt limphoma ) cell lines. Dacarbazine and methotrexate were used for comparative purpose. The 3-methyl-4-(3,3-dimethyltriazeno)-5-(substituted benzamido)pyrazoles, bearing the pyrazole nucleus free at 1 position, resulted more active than the 1-(substituted phenyl)-3-methyl-4-(3,3-dimethyltriazeno)-5 benzamidopyrazoles. Dacarbazine at 10 microM showed no activity in the above tests. The observed difference among Dacarbazine and the active 4 triazenopyrazoles migth be explained admiting that these last compounds, differently by Dacarbazine, did not follow a mechanism of action based on the cytochrome P-450 induced demethylation. The most active compound 2d showed growth inhibition values of 97.8 and 99.4% against K562 and Raji cell lines respectively. Methotrexate inhibition values at 0.2 microM against the above cell lines were 86.7 and 75.1% respectively. PMID- 15120322 TI - Comparison of capillary zone electrophoresis and high performance liquid chromatography methods for quantitative determination of ketoconazole in drug formulations. AB - A capillary zone electrophoresis (CZE) and a high-performance liquid chromatography (HPLC) method have been developed for identification and determination of ketoconazole, an imidazole antifungal, in pharmaceutical preparations. The suitabilities of both methods for quantitative determination of ketoconazole were approved through validation specification such as linearity, precision, accuracy, limit of detection and quantification. The proposed methods were used for determination of ketoconazole in commercial pharmaceutical dosage forms (tablets and creams). Under described conditions, CZE method is more selective, while the HPLC method is more sensitive. Both methods are rapid (tR(CZE)=5.14 min and tR(HPLC)=2.66 min), which is important for routine application. However, the HPLC method provides a repeatability of the quantitative analysis of ketoconazole in drug formulations below 1.5% relative standard deviation (R.S.D), while the repeatability of the CZE method is in the order of 2-3% R.S.D. PMID- 15120323 TI - Comparison of zero- and second-order derivative spectrophotometric and HPLC methods for the determination of gemcitabine in human plasma. AB - Zero- and second-order derivative spectrophotometric and high-performance liquid chromatography (HPLC) methods were developed and validated for the determination of gemcitabine in human plasma. Spectrophotometrically, gemcitabine was determined by means of zero-order derivative absorbance values (A) at 288 nm and from values from the second-order derivative absorbance values (2D) at 285 nm. Beer's Law was obeyed in the range 0.50-15.0 microg ml(-1). The proposed other method, normal-phase HPLC method for determination of gemcitabine in human plasma was described. Calibration curve was linear over the concentration range 0.20 15.0 microg ml(-1). Quantitation was achieved by diode array detection at 272 nm using 2'-deoxycytidine as internal standard. Results obtained by spectrophotometric and HPLC methods for determination of gemcitabine in human plasma described in this paper showed adequate accuracy, precision and repeatability. No interference was found in plasma at the selected derivative wavelength and chromatographic conditions. According to the statistical comparison, there is no significant difference between the three methods. This is suggested that the three methods are equally applicable. PMID- 15120324 TI - Stringent rosiglitazone-dependent gene switch in muscle cells without effect on myogenic differentiation. AB - We have developed a gene switch based on the human transcription factor peroxisome proliferator-activated receptor gamma (PPARgamma) and its activation by rosiglitazone. However, ectopic expression of PPARgamma has been demonstrated to convert myogenic cells into adipocyte-like cells and, more generally, may interfere with the physiology of the target tissue. Consequently we modified the DNA-binding specificity of PPARgamma, resulting in a transcription factor that we named PPAR*. We demonstrated by histological and molecular assessment of cell phenotype that the overexpression of PPAR* did not alter the myogenic differentiation program of G8 myoblasts. We showed that PPAR* does not transactivate promoters containing PPARgamma-responsive elements but transactivates promoters containing PPAR*-responsive elements that are at least 80% identical to a 20-bp consensus. We improved the rosiglitazone-dependent gene switch by tuning PPAR* expression with a scaffold/matrix attachment region and by expressing both PPAR* and the reporter gene under the control of PPAR*-responsive elements. Treatment of cultured murine muscle cells (myotubes) with rosiglitazone induced reporter gene expression from assay background up to the level attained by a CMV I/E promoter-enhancer. These results indicate the potential of the PPAR* gene switch for use in gene therapy applications. PMID- 15120325 TI - Xenograft models for liver metastasis: Relationship between tumor morphology and adenovirus vector transduction. AB - The improvement of initial tumor cell transduction with viral vectors is a major task in tumor gene therapy. We have developed mouse tumor models with hepatic metastases to study transduction of tumor cells after systemic adenovirus vector application. The tumor models were established by intraportal transplantation of human tumor cell lines into immunodeficient mice. Liver metastases derived from cervix, colon, breast, and liver cancer lines were analyzed for distribution of extracellular matrix, vascularization, and transgene expression after tail vein injection of adenovirus vectors. Overall, xenografts resembled the morphology of corresponding tumors in cancer patients. Adenovirus-mediated gene delivery depended on tumor vascularization and direct contact between blood vessels and tumor cells. These models represent important tools for studying and improving tumor gene therapy approaches. PMID- 15120326 TI - Destruction of nonimmunogenic mammary tumor cells by a fusogenic oncolytic herpes simplex virus induces potent antitumor immunity. AB - In principle, destruction of tumor cells in vivo by oncolytic agents would release the entire repertoire of tumor antigens in their natural forms, leading to effective antitumor immunity. This goal has been elusive despite extensive testing of numerous strategies. We developed a doubly fusogenic oncolytic herpes simplex virus (Synco-2D) that kills tumor cells by a unique dual mechanism combining direct cytolysis with syncytial formation induced by cell membrane fusion. A single intratumor injection of Synco-2D induced strong antitumor immunity against an otherwise nonimmunogenic murine mammary tumor growing in immune-competent mice. CD8+ T cells were the primary mediators of immunity, contributing to the destruction of both primary and metastatic tumors. We conclude that the fusogenic capacity of Synco-2D enables it to elicit antitumor immunity exceeding that induced by more conventional oncolytic viruses. PMID- 15120327 TI - Overcoming acquired resistance to TRAIL by chemotherapeutic agents and calpain inhibitor I through distinct mechanisms. AB - We recently found that repeated application of adenovectors expressing the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) or recombinant TRAIL proteins to TRAIL-susceptible cancer cells resulted in selection and expansion of TRAIL-resistant cells. Overcoming this acquired resistance to TRAIL is desirable for TRAIL-mediated cancer therapy. Here we demonstrate that several chemotherapeutic agents, including 5-fluorouracil (5-FU) and mitomycin, and calpain inhibitor I, an NFkappaB inhibitor, can overcome acquired resistance to TRAIL in DLD1 colon cancer cells. The combination of TRAIL (approved gene symbol TNFSF10) gene therapy and 5-FU enhanced tumor suppression in vivo in nude mice bearing subcutaneous tumors established from TRAIL-resistant colon cancer cells. Whereas treatment with the combination of TRAIL and 5-FU or mitomycin led to enhanced activation of caspase-3, the combination of TRAIL and calpain inhibitor I resulted in enhanced activation of both caspase-8 and caspase-3. Moreover, mitomycin, but not 5-FU or calpain inhibitor I, induced overexpression of the BAX gene, which was correlated with enhanced TRAIL-induced cell killing in TRAIL resistant DLD1 cells. Together, these results suggest that acquired resistance to TRAIL can be overcome by different mechanisms and that combinations of TRAIL gene therapy and chemotherapy may be a useful approach for cancer treatment. PMID- 15120328 TI - Induction of potent TRAIL-mediated tumoricidal activity by hFLEX/Furin/TRAIL recombinant DNA construct. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been shown to exert selectively cytotoxic activity against many tumor cells but not normal cells. On the other hand, the ligand for the receptor tyrosine kinase Fms-like tyrosine kinase 3 (Flt3L) is a growth factor for hematopoietic progenitors and is a potent stimulating factor for dendritic and NK cells. Previously, we have demonstrated that it is possible to inhibit the outgrowth of primary tumors by the administration of an hFlex (the extracellular domain of the Flt3L) and TRAIL (amino acid residues 95-281) secreted fusion protein. Here, we report that by the insertion of a linker sequence encoding the cleavage site for the Golgi-expressed endoprotease furin between the DNA sequences encoding hFlex and TRAIL, the tumoricidal activity of the cleaved TRAIL protein generated was greatly enhanced in comparison to the hFlex/TRAIL fusion protein. Furthermore, we demonstrate that intratumoral injection of the hFlex/furin/TRAIL DNA, in conjunction with cationic liposomes, significantly suppressed the outgrowth of the human CNE-2 nasopharyngeal tumor xenografts in SCID mice. In situ histological examinations confirmed the expression of TRAIL in the treated tumor nodules and the induction of apoptosis was also evidenced by the presence of numerous pyknotic nuclei. PMID- 15120329 TI - AAV-mediated gene delivery of BDNF or GDNF is neuroprotective in a model of Huntington disease. AB - Huntington disease (HD) is a neurodegenerative disorder that results in the progressive loss of GABAergic medium spiny projection neurons in the striatum. Neurotrophic factors have demonstrated neuroprotective actions on striatal neurons, suggesting that increased neurotrophic factor expression may prevent or reduce neuronal loss in the HD brain. We investigated whether enhanced expression of brain-derived neurotrophic factor (BDNF) or glial cell line-derived neurotrophic factor (GDNF), achieved by adeno-associated viral (AAV) vector mediated gene delivery, could protect striatal neurons in the quinolinic acid (QA) rodent model of HD. Adult Wistar rats received unilateral intrastriatal injections of AAV-BDNF, AAV-GDNF, AAV-GFP, or PBS. Three weeks later, the rats were lesioned with QA, a toxin that induces striatal neuron death by an excitotoxic process. Both AAV-BDNF and AAV-GDNF significantly reduced the loss of both NeuN- and calbindin-immunopositive striatal neurons 2 weeks after lesion compared to controls. AAV-BDNF also provided significant neurotrophic support to NOS-immunopositive striatal interneurons, while AAV-GDNF-treated rats demonstrated significant protection of parvalbumin-immunopositive striatal interneurons compared to controls. These results indicate that AAV-mediated gene transfer of BDNF or GDNF into the striatum provides neuronal protection in a rodent model of HD. PMID- 15120330 TI - Overexpression of p53 increases lumen size and blocks neointima formation in porcine interposition vein grafts. AB - Patency rates for autologous saphenous vein (SV) conduits used in coronary artery bypass grafts remain poor. Patients with failed grafts are difficult to treat with subsequent interventions, necessitating the development of innovative therapies. Previous studies have suggested that induction of smooth muscle cell (SMC) apoptosis may reduce neointima formation. We overexpressed the proapoptotic gene p53 at the lumenal surface of SV grafts using adenoviral (Ad)-mediated gene transfer in porcine SVs prior to grafting in vivo and analyzed at 7 and 28 days (n = 6 and 7 per group, respectively). p53 overexpression induced a significant upregulation in apoptosis (4 +/- 0.6% for Adp53-infected grafts vs 0.6 +/- 0.1% for Adbeta-gal-infected grafts) and reduced neointimal proliferation by 28 +/- 1% at day 7 postinfection. Adp53-infected grafts had significantly greater lumenal areas than controls at both time points (4.8 +/- 0.6 mm2 vs 2.9 +/- 0.5 mm2 and 10.0 +/- 2.5 mm2 vs 4.2 +/- 1.2 mm2 at 7 and 28 days, respectively). Total graft areas were also increased at 28 days by p53, indicating positive vessel remodeling. Additionally, the thickening of the neointima was significantly reduced by 68 +/- 22% and 28 +/- 3% by p53 overexpression at day 7 and 28, respectively. Importantly, phenotypic changes were maintained at 3 months. Induction of SMC apoptosis by transient p53 overexpression positively influenced vein graft remodeling. PMID- 15120331 TI - Adenovirus encoding human platelet-derived growth factor-B delivered in collagen exhibits safety, biodistribution, and immunogenicity profiles favorable for clinical use. AB - We have developed a therapeutic approach to wound repair involving immobilization of gene transfer vectors within biocompatible matrices (gene-activated matrix, or GAM). The matrix also serves as a scaffold for cellular in-growth and subsequent gene uptake and expression. An adenoviral vector encoding human platelet-derived growth factor-B delivered in collagen (AdPDGF-B/GAM) has demonstrated efficacy in models of wound repair. The safety, biodistribution, and immunogenicity profiles of AdPDGF-B/GAM were examined using a rabbit dermal wound model. Four weekly doses at 1 x 10(10) and 1 x 10(11) viral particles/cm2 of wound surface stimulated dose-related increases in granulation tissue formation and cell proliferation. In situ hybridization and immunostaining demonstrated concordant expression of human PDGF-B mRNA and protein. No treatment-related changes in hematology, serum chemistry, or histopathology were observed. Although AdPDGF-B DNA and PDGF-B mRNA were detected in wounds and axillary lymph nodes of treated animals, no AdPDGF-B was detected in blood or other organs. No immunologic responses against collagen were observed; however, as expected, IgG responses to AdPDGF-B and human PDGF-BB protein were detected. In adenovirus-preimmunized rats, attenuation of the wound healing response was modest (approximately 16%). Collectively, these observations indicate that repeat doses of AdPDGF-B/GAM are well tolerated and lead to robust, localized tissue repair. PMID- 15120333 TI - Intracellular delivery of functional proteins via decoration with transporter peptides. AB - Despite numerous attractive intracellular targets, protein therapeutics have been principally confined to the extracellular space due to the lack of a straightforward way to deliver functional polypeptides to the cell interior. Peptide sequences facilitating intracellular protein delivery have been identified; however, current strategies to apply them require problematic steps, such as generation of new in-frame fusion proteins, covalent chemical conjugation, and denaturation. We have developed a new approach to protein transfer into cells and tissues that relies on single-step decoration by cysteine flanked, arginine-rich transporter peptides. This approach facilitated cell and tissue delivery of a variety of functional proteins, including antibodies and enzymes. Decoration with transporter peptides thus provides an attractive general means of intracellular delivery of functional proteins in vitro and in tissue. PMID- 15120332 TI - Enhanced gene transfer to mouse dendritic cells using adenoviral vectors coated with a novel adapter molecule. AB - Adenovirus (Ad)-mediated transduction of dendritic cells (DC) is inefficient because of the lack of the primary Ad receptor, CAR. DC infection with Ad targeted to the CD40 results in increased gene transfer. The current report describes further development of the CD40-targeting approach using an adapter molecule that bridges the fiber of the Ad5 to CD40 on mouse DC. The adapter molecule, CFm40L, consists of CAR fused to mouse CD40 ligand via a trimerization motif. A stable cell line that secretes CFm40L at high levels was generated. Gene transfer to mouse bone marrow-derived DC (mBMDC) using CFm40L-targeted Ad was over 4 orders of magnitude more efficient than that for the untargeted Ad5. Gene transfer was achieved to over 70% of the mBMDC compared to undetectable transduction using untargeted Ad5. In addition to dramatically enhanced gene transfer, the CFm40L-targeted Ad5 induced phenotypical maturation and upregulated IL-12 expression. Most importantly, the CFm40L-targeted Ad5 elicited specific immune response against a model antigen in vivo. The results of this study demonstrate that Ad-mediated gene transfer to DC can be significantly enhanced using nonnative transduction pathways, such the CD40 pathway, which may have important applications in genetic vaccination for cancer and infectious diseases. PMID- 15120334 TI - Effects of microtubule-depolymerizing agents on the transfection of cultured vascular smooth muscle cells: enhanced expression with free drug and especially with drug-gene lipoplexes. AB - The microtubule-depolymerizing agents colchicine, vinblastine (VB), vincristine, nocodazole, and podophyllotoxin were found to increase dramatically the transfection of cationic phospholipid-DNA (CMV-beta-gal) complexes on cultured vascular smooth muscle cells (VSMCs). Pretreatment of cells with free colchicine before addition of lipoplexes increased transgene expression both in the presence and in the absence of serum. Free vinblastine had similar effects; however, vinblastine was more effective (approximately 30-fold maximal stimulation) when incorporated into the lipoplexes. Under optimal conditions, vincristine, nocodazole, and podophyllotoxin produced 25- and 39-, 31- and 14-, and 26- and 14 fold increases in the absence and presence of serum, respectively. Taxol, which stabilizes microtubules, had no effect on transfection, but it blocked the positive effect of colchicine. Cytochalasin B, which inhibits microfilament polymerization, had no effect on transgene expression. By fluorescence microscopy, normal lipoplexes colocalized with lysosomes. In contrast, there was little, if any, colocalization of VB lipoplexes with lysosomes. Because depolymerization of microtubules induces NF-kappaB-dependent gene expression, the effects of pyrrolidinedithiocarbamate and Nalpha-p-tosyl-L-lysine chloromethyl ketone, inhibitors of NF-kappaB activation, were tested; inhibition of vinblastine stimulation of transfection was 85 and 66%, respectively. Also, immunofluorescence microscopy showed that vinblastine induced the translocation of NF-kappaB from the cytoplasm to the nucleus. It is concluded that microtubule depolymerizing agents, especially when incorporated into lipoplexes, dramatically increase transfection of VSMCs, probably by two mechanisms: (i) inhibition of transport of lipoplexes to lysosomes and (ii) activation of transcription (via NF kappaB). There have been some reports on the use of pharmaceutical agents to enhance gene expression, but generally these have involved separate applications of drug and gene. The ability to deliver a drug and a gene in a single therapeutic formulation could have significant clinical implications. PMID- 15120335 TI - Simplified generation of high-titer retrovirus producer cells for clinically relevant retroviral vectors by reversible inclusion of a lox-P-flanked marker gene. AB - Retroviral producer cells are generated by the introduction of a viral genome into "helper" cell lines containing all the necessary components for viral packaging and the release of infectious particles. The selection of high-titer vector producer cells is most efficient if the vector genome encodes a selectable marker, while it is extremely tedious to select high-titer producer clones if the transgene cannot be detected and selected directly. Here we describe the development of a screening system that uses reversible integration of lox-P flanked eGFP as a qualitative and quantitative marker gene in two different vector systems, greatly facilitating the selection of viral producer cells. After selection and titration of high-titer viral producer cells based on eGFP expression, the eGFP gene could be removed from the provirus by transient introduction of Cre-recombinase into the producer cells, thus allowing the production of therapeutic relevant vectors expressing solely the gene of interest. However, after removal of the marker gene a slight but consistent increase in viral titers compared to the respective control vectors was found, independent of the transgene or backbone used. The single lox-P site retained in the vector backbone does not affect gene expression level or fidelity of RNA processing. PMID- 15120336 TI - Internalized antigens must be removed to prepare hypoimmunogenic mesenchymal stem cells for cell and gene therapy. AB - Adult stem cells from human bone marrow stroma, referred to as mesenchymal stem cells or marrow stromal cells (hMSCs), are attractive candidates for clinical use. The optimal conditions for hMSC expansion require medium supplemented with fetal calf serum (FCS). Some forms of cell therapy will involve multiple doses, raising a concern over immunological reactions caused by medium-derived FCS proteins. By a sensitive fluorescence-based assay we determined that 7 to 30 mg of FCS proteins are associated with a standard preparation of 100 million hMSCs, a dosage that probably will be needed for clinical therapies. Here we present ex vivo growth conditions for hMSCs that reduce the FCS proteins to less than 100 ng per 100 million hMSCs, approximately a 100,000-fold reduction. The cells maintain their proliferative capacity and sustain their ability for multilineage differentiation. Experiments in rats demonstrate that rat MSCs grown in 20% FCS induce a substantial humoral response after repeated administrations, whereas cells grown under the conditions described in this study reduce the immunogenicity in terms of IgG response over 1000-fold to barely detectable levels. Our results have the potential to dramatically improve cellular and genetic therapies using hMSCs and perhaps other cells. PMID- 15120337 TI - Transfection of RNA encoding tumor antigens following maturation of dendritic cells leads to prolonged presentation of antigen and the generation of high affinity tumor-reactive cytotoxic T lymphocytes. AB - Common tumor vaccination strategies utilizing peptide-pulsed dendritic cells (DC) are limited to targeting antigens with known epitopes in patients expressing a defined restricting allele and can result in the preferential induction of low avidity T cells that fail to recognize tumor cells. The use of dendritic cells transfected with RNA encoding tumor antigen offers the prospect of antigen specific immunization without requiring prior knowledge of the immunogenic epitope or restricting allele, since epitopes from the translated protein are processed by the endogenous antigen-presentation machinery. However, its use in vaccine studies has been limited by low RNA transfection efficiency and the use of immature DC as recipient cells. In this study, we report an RNA transfection strategy that routinely achieves expression in 40-50% of mature DC, which are better stimulator cells. Such RNA-transfected mature DC exhibited a prolonged duration of presentation of immunogenic epitopes compared to peptide-pulsed DC, induced greater frequencies of tumor antigen-specific CTL, and generated a CTL population that exhibited higher target avidity and increased tumor lytic capacity. These studies provide compelling in vitro data supporting the evaluation of RNA-transfected mature DC in vaccination protocols as a means to overcome several obstacles to generating anti-tumor responses in vivo. PMID- 15120338 TI - Injecting reason. PMID- 15120339 TI - Influenza: time to come to grips with the avian dimension. PMID- 15120340 TI - Polio eradication-rethinking the endpoint of the end game. PMID- 15120341 TI - Development of genetically modified mosquitoes in Africa. PMID- 15120342 TI - Transmission of HIV-2: another perspective. PMID- 15120343 TI - Vulnerability to malaria, tuberculosis, and HIV/AIDS infection and disease. Part 1: determinants operating at individual and household level. AB - A high burden of malaria, tuberculosis, and HIV infection contributes to national and individual poverty. We have reviewed a broad range of evidence detailing factors at individual, household, and community levels that influence vulnerability to malaria, tuberculosis, and HIV infection and used this evidence to identify strategies that could improve resilience to these diseases. This first part of the review explores the concept of vulnerability to infectious diseases and examines how age, sex, and genetics can influence the biological response to malaria, tuberculosis, and HIV infection. We highlight factors that influence processes such as poverty, livelihoods, gender discrepancies, and knowledge acquisition and provide examples of how approaches to altering these processes may have a simultaneous effect on all three diseases. PMID- 15120344 TI - Mucosa or skin as source of coagulase-negative staphylococcal bacteraemia? AB - Nosocomial bacteraemia is associated with significant morbidity, mortality, and cost worldwide, and is most commonly caused by coagulase-negative staphylococci (CONS). Establishing the source of CONS bacteraemia is therefore important in the prevention and management of this infection. CONS infections are presumed to originate at the cutaneous sites of central venous catheters (CVCs), a belief that has led to prevention strategies that focus almost exclusively on the skin. However, mucosal colonisation by CONS is well established, suggesting that mucosal sites might be an important source of CONS bacteraemia. We review the published material that evaluates the source(s) of CONS. We included only studies that used a strict definition of CONS bacteraemia, evaluated skin and other potential sources of CONS, and studied the molecular association between CONS blood isolates and their potential sources. Three published reports fulfilled our criteria. In cancer patients with CONS or CONS bacteraemia, most of the colonising strains that had a molecular match with the strain recovered from the blood of the same patient were mucosal isolates; by contrast, no association was seen between CONS blood and skin isolates. Furthermore, in several patient populations evidence was reported of mucosal colonisation by CONS and in several reports experimental and clinical mucosal translocation of CONS with subsequent bacteraemia was documented. Together these data indicate that mucosal sites are an important source of CONS bacteraemia. Clinical strategies for the treatment of patients with a positive blood culture for CONS, the widespread use of antimicrobial-coated CVCs, and maximum barrier protection for CVC insertion should be reassessed, and strategies to decrease mucosal colonisation by CONS should be developed. PMID- 15120345 TI - Tuberculosis in New York city: recent lessons and a look ahead. AB - In the late 1980s and early 1990s, after decades of decline, the incidence of tuberculosis began to rise in New York city, reaching a peak of 3811 cases by 1992. The epidemic took root in a setting of inadequate treatment regimens, homelessness, a diminished public-health system, and the onset of the HIV/AIDS epidemic. In addition, a subepidemic of drug-resistant tuberculosis occurred throughout New York city, most notably in a series of well documented nosocomial outbreaks. By 1994, using broadened initial treatment regimens, directly observed therapy, and improved US Centers for Disease Control and Prevention guidelines for hospital control and disease prevention, New York city began to effectively halt the progression of the epidemic. By 2002, tuberculosis rates in New York city reached an historic low of 1084. However, given the presence of a large reservoir of latently infected individuals in the city and an ongoing tuberculosis pandemic, New York city continues to face significant challenges from this persistent pathogen. PMID- 15120346 TI - Infections related to the ingestion of seafood. Part II: parasitic infections and food safety. AB - Parasites are responsible for a substantial number of seafood-associated infections. The factor most commonly associated with infection is consumption of raw or undercooked seafood. People with underlying disorders, particularly liver disease, are more susceptible to infection. In the first part of this review, published last month, we discussed the viral and bacterial agents associated with consumption of seafood. In part II, we discuss the parasites commonly associated with seafood consumption. Parasites readily identifiable from both consumable seafood and infected human beings include nematodes, trematodes, cestodes, and protozoa. The salient features associated with seafood-related parasite infestations are discussed. To provide a safe product for consumers, the seafood industry and the government in the USA have undertaken specific measures, which include good manufacturing practices and hazards analysis and critical control points implemented by the government and regulatory agencies. Consumers should take common precautions including obtaining seafood from reputable sources especially if the seafood is to be consumed uncooked. Adequate cooking of seafood is the safest way of preventing related infections. PMID- 15120347 TI - Rationale for evaluating North America's first medically supervised safer injecting facility. AB - Many cities throughout the world are experiencing ongoing infectious disease and overdose epidemics among illicit injection drug users (IDUs). In particular, HIV and hepatitis C virus (HCV) have become endemic in many settings and bacterial infections, such as endocarditis, have become extremely common among this population. In an effort to reduce these public health concerns, in September 2003, Vancouver, Canada, opened a pilot medically supervised safer- injecting facility (SIF), where IDUs can inject pre-obtained illicit drugs under the supervision of medical staff. Before and since the facility's opening, there has been a substantial misunderstanding about the rationale for evaluating SIF as a public-health strategy. This article outlines the evidence and rationale in support of the Canadian initiative. This rationale involves limitations in conventionally applied drug-control efforts, and gaps in current public-health policies in controlling the spread of infectious diseases, and the incidence of overdose among IDUs. PMID- 15120349 TI - Marc Struelens--President of the European Society of Clinical Microbiology and Infectious Diseases. Interviewed by Pam Das. PMID- 15120351 TI - Technology and public health. PMID- 15120352 TI - Must be something in the water. PMID- 15120355 TI - CT diagnosis of postoperative intussusception after penetrating abdominal trauma. PMID- 15120354 TI - Proctocolectomy with ileoanal anastomosis. PMID- 15120357 TI - Open Roux-en-Y gastric bypass: indications and technique. PMID- 15120358 TI - Open vs. laparoscopic procedures in bariatric surgery. PMID- 15120359 TI - Controversies in bariatric surgery: evidence-based discussions on laparoscopic adjustable gastric banding. AB - Laparoscopic adjustable gastric banding (LAGB) is a surgical option that involves placing a silicone band circumferentially around the uppermost aspect of the stomach. The band creates a small proximal pouch that empties slowly resulting in early satiety and a decreased appetite. The band is attached to an access port that is secured to the rectus muscle and can be accessed percutaneously in the office with a needle. Injection of saline into the port results in tightening of the band. This is performed on an individual basis according to weight loss and appetite. Band adjustments are required approximately 5-6 times in the first year and 2-3 times in the second year. Weight loss is gradual, averaging 1-2 lb/week during the first 2 years after surgery. PMID- 15120360 TI - Roux-en-Y gastric bypass is the operation of choice for bariatric surgery. PMID- 15120361 TI - Is gastric bypass superior for the surgical treatment of obesity compared with malabsorptive procedures? PMID- 15120363 TI - Biliopancreatic diversion with duodenal switch vs. gastric bypass for severe obesity. PMID- 15120364 TI - Implantable gastric stimulation for weight loss. AB - With the epidemic of obesity worldwide, bariatric surgery has rapidly grown in popularity. Currently, a variety of surgical procedures are performed including Roux-en-Y gastric bypass, laparoscopic adjustable gastric banding, vertical banded gastroplasty, and biliopancreatic diversion. All of these procedures have been shown to succeed in achieving significant and sustainable weight loss for the majority of patients. However, these procedures also carry the potential for serious operative morbidity, altered gastrointestinal anatomy or function, or both. Electrical gastric stimulation via the implantable gastric stimulation (IGS) system is a relatively new and novel approach to treat obesity. The operative technique is relatively simple and the system does not alter gastrointestinal anatomy. Preliminary worldwide investigations have demonstrated safety and efficacy. This article will review the current experience with the IGS system. PMID- 15120365 TI - Role of retinoid X receptor mRNA expression in Barrett's esophagus. AB - The Barrett's multistage process is characterized histopathologically by progression from Barrett's intestinal metaplasia to Barrett's esophagus with dysplasia and ultimately adenocarcinoma. Understanding of the molecular alterations in this multistage process may contribute to improved diagnosis and treatment. Retinoid X receptors (RXR) play an important role in regulating the morphogenesis, development, growth, and differentiation of cells. Alterations in RXR expression have been observed in a variety of solid tumors; however, the role in Barrett's esophagus disease has yet to be determined. The aim of this study was to assess the prevalence and timing of RXR messenger RNA expression in the Barrett's metaplasia-dysplasia-adenocarcinoma sequence and to investigate its role in the development and progression of this disease. We analyzed the mRNA expression of all three RXR subtypes (RXR-alpha, RXR-beta, and RXR-gamma) by using a quantitative real-time reverse transcription-polymerase chain reaction method in 108 specimens from 19 patients with Barrett's esophagus without carcinoma (BE group), 20 patients with Barrett's-associated adenocarcinoma (EA group), and a control group of 10 patients without evidence of gastroesophageal reflux disease (CG). RXR-alpha mRNA expression was significantly decreased (P < 0.001; Kruskal-Wallis test), and RXR-gamma was significantly increased (P < 0.001) at higher stages in Barrett's esophagus. RXR-beta expression was highest in Barrett's tissues and was significantly increased compared to normal squamous tissues (P=0.01; Wilcoxon test) and adenocarcinoma tissues (P=0.018, Mann-Whitney test). RXR-alpha and RXR-beta mRNA expression was significantly associated in normal squamous esophagus tissues (r(2)=0.49; P < 0.001; Spearman test), Barrett's tissues (r(2)=0.63; P < 0.001), and adenocarcinoma tissues (r(2)=0.68; P=0.001). There were significant differences in RXR-alpha (P=0.011) and RXR-beta (P=0.005) mRNA expression in histopathologically normal squamous esophagus tissues in patients with cancer and the control group without evidence of gastroesophageal reflux disease. These findings suggest that alterations in the mRNA expression of all three RXR subtypes are frequent events in the development and progression of Barrett's esophagus and associated adenocarcinoma, that RXR mRNA expression levels may be useful biomarkers for this disease, and that a widespread "field-effect" is present in the normal esophagus of patients with esophageal adenocarcinoma. PMID- 15120366 TI - GSTP1, GSTM1, and GSTT1 genetic polymorphisms in patients with cryptogenic liver cirrhosis. AB - We investigated glutathione S-transferase (GST) P1 Ile (105) Val, T1, and M1 polymorphisms in 45 patients with documented cryptogenic cirrhosis and 56 healthy control subjects. Polymerase chain reaction-based procedures were performed in the studied populations to confirm the genotypes of GSTT1, M1, and P1. Ile/Val and Val/Val GSTP1 genotypes were more frequent in the patients with cirrhosis (n=39, 87%) than in the control subjects (n=10; 18%) (odds ratio [OR] 34.04; 95% confidence interval [CI] 10.70 to 108.31, P<0.001). Among these patients with cirrhosis, 16 were heterozygous and 23 were homozygous, whereas only one person in the control group was homozygous. The GSTM1 null genotype was also more prevalent in cirrhotic patients than in healthy control subjects (OR 6.83, 95% CI 2.53 to 18.42, P<0.001). The rate of GSTT1 deletion did not show a significant difference between the two groups (OR 2.35, 95% CI 0.76 to 7.28, P=0.111). To our knowledge, this is the first evidence that GSTP1 and GSTM1 polymorphisms may be related to the development of cirrhosis by unknown mechanisms. The significant association of cryptogenic cirrhosis with Val/Val GSTP1 genotype encoding a low detoxification activity protein implicates this polymorphism as a risk factor for the occurrence of the disease. PMID- 15120367 TI - Postoperative jejunal feeding and outcome of pancreaticoduodenectomy. AB - Complications following pancreaticoduodenectomy are common, partly because of nutritional debilitation. The aim of this study was to evaluate the impact of early postoperative tube feeding on outcome of pancreaticoduodenectomy and determine the best method for delivering enteral feeding. A retrospective review of 180 consecutive patients undergoing Whipple operations from 1994 to 2000 was performed. Two nonrandomized patient groups were retrospectively studied: those with early postoperative tube feeding vs. those with no planned feeding. Ninety eight patients (54%) received postoperative jejunal feeding, whereas 82 patients (46%) did not. Jejunal feeding was delivered via a bridled nasojejunal tube in 55 patients (56%) and a gastrojejunal tube in 43 (44%). Vomiting (10% vs. 29%; P=0.002) and use of total parenteral nutrition (6% vs. 27%; P < 0.0001) were less in the jejunal feeding group as well as rates of readmission (12% vs. 27%; P=0.022), early (52% vs. 62%; P=0.223) and late (12% vs. 31%, P=0.005) complications, and infections (13% vs. 20%, P=0.014). Tube-related complications occurred in 6 of 98 patients, all of which were associated with gastrojejunal tubes (P=0.021). Early postoperative tube feeding after pancreaticoduodenectomy is associated with significantly less use of total parenteral nutrition and lower rates of readmission and complications. A bridled nasojejunal feeding tube appears to be a safe and reliable method of short-term enteral feeding. PMID- 15120369 TI - Mechanical consequences of short gastric vessel division at the time of laparoscopic total fundoplication. AB - Laparoscopic Nissen fundoplication is currently the most commonly practiced antireflux operation. Some adverse consequences of the operation remain in the form of mechanical side effects, labeled postfundoplication complaints, of which dysphagia and gas bloat seem to predominate. Measures have been suggested to counteract some of these and one frequently advocated has been division of the short gastric vessels to create a short-floppy wrap. The advantages of this are still debated, particularly in the long-term perspective. The aim of the present study was to evaluate the mechanical consequences of dividing all short gastric vessels at the time of a laparoscopic total fundoplication. Ninety-nine patients with chronic gastroesophageal reflux disease (GERD) were originally allocated on a random basis to have either all short gastric vessels divided or left intact at the time of a laparoscopic total fundoplication. A subsample of these patients, again selected at random, were recruited for a comprehensive manometric investigation 1 year after the operation. In this cohort, 12 patients had all short gastrics divided and in 12 patients, the wrap was done with intact vessels by use of the anterior portion of the fundus. Manometry was carried out by the use of a sleeve sensor to straddle the lower esophageal sphincter (LES), and gastric distension (750 ml air) was used to trigger transient LES relaxations (TLESR). The basal LES tone was similar in the two groups (14.2 +/- 2.4 and 18.8 +/- 4.3, mean +/- SE), respectively. Accordingly, all other relevant manometric variables were equal when the two groups were compared, except for the total number of TLESRs (triggered by gastric distension by air) that were significantly higher (p < 0.02) in patients having their short gastric vessels intact. Consequently, numerically more common cavities were recorded in the latter group. Very similar outcomes in terms of motor function of the LES and esophageal body were observed after a total fundoplication irrespective of whether a complete division of all gastric vessels had been carried out or not. However, after gastric distension with air, more TLESRs were recorded in the latter group suggesting a better maintained ability to vent air from the stomach. PMID- 15120368 TI - Adenocarcinoma appearing very late after antireflux surgery for Barrett's esophagus: long-term follow-up, review of the literature, and addition of six patients. AB - Antireflux surgery is supposed to prevent the development of adenocarcinoma in patients with Barrett's esophagus. The purpose of this study was to determine the prevalence of adenocarcinoma late after antireflux surgery. A total of 161 patients with long-segment Barrett's esophagus had antireflux surgery and were followed for a mean of 148 months (range 54 to 268 months) Clinical, endoscopic, histologic, and functional studies were performed. Of the original 161 patients, 147(91.3%) completed long-term follow-up. Six patients (4.1%) developed adenocarcinoma 4,5,6,9,17, and 18 years, respectively, after surgery. Five were men. Two of them were asymptomatic for 12 and 17 years. Three of them had extra long-segment Barrett's esophagus. Five underwent manometric evaluation with only one showing an incompetent lower esophageal sphincter. In two cases, 24-hour pH studies showed massive acid reflux. Two patients had early adenocarcinoma, whereas four had advanced carcinoma. Adenocarcinoma in long-segment Barrett's esophagus seems to develop mainly in patients with recurrence of pathologic reflux, especially among men. A review of the English language literature during the last 23 years found 25 articles dealing with Barrett's esophagus and antireflux surgery. Most of these reports had only a few patients with short-term follow-up (<60 months). To determine the true prevalence of this complication, a long-term objective follow-up is necessary. PMID- 15120370 TI - Epidermal growth factor receptor expression correlates with histologic grade in resected esophageal adenocarcinoma. AB - Activation of the epidermal growth factor receptor (EGFR) has a role in oncogenesis and may correlate with prognosis. The aim of this study was to examine EGFR expression in esophageal adenocarcinoma and correlate EGFR status with pathologic and clinical prognostic features. An exploratory retrospective review of 38 patients with surgically resected esophageal adenocarcinoma was performed. All patients underwent an esophagogastrectomy with regional lymphadenectomy; 24 patients underwent primary resection and 14 patients had surgery after preoperative chemoradiation therapy. Immunohistochemical analysis was performed on paraffin-embedded tissue samples using an EGFR monoclonal antibody. Low- and moderate-grade tumors were positive for EGFR expression in 2 of 15 patients; poorly differentiated tumors were positive for EGFR expression in 13 of 23 patients (p=0.02). The median survival was 35 months (confidence interval [CI]: 29-40) for EGFR negative patients (n=23) and 16 months (CI: 10-22) for EGFR positive patients (n=13) (p=0.10). Disease recurred in 3 of 21 EGFR negative patients and 6 of 13 EGFR positive patients (p=0.06). Poorly differentiated adenocarcinomas of the esophagus demonstrated higher EGFR expression compared to low-grade tumors based upon immunohistochemical analysis. A trend toward improved disease-free and overall survival was seen in EGFR negative patients. PMID- 15120371 TI - Cytoreductive surgery with intraperitoneal hyperthermic chemotherapy for advanced gastric cancer. AB - Peritoneal carcinomatosis is a common and universally fatal sequelae of gastric carcinoma. Treatment of peritoneal carcinomatosis from appendiceal and colorectal sources with intraperitoneal hyperthermic chemotherapy (IPHC) combined with aggressive cytoreductive surgery has been shown to be effective. There are few data on this treatment modality for carcinoma of the stomach. This study evaluates cytoreductive surgery and IPHC with peritoneal carcinomatosis from gastric carcinoma. Thirty-four patients with peritoneal carcinomatosis due to gastric carcinoma underwent gastric resection with cytoreductive surgery followed by IPHC with mitomycin C. A control group consisting of 40 contemporaneous patients, who underwent radical gastrectomy without extended nodal resection, was identified through the tumor registry. Despite more advanced disease in the IPHC group compared to the control group (P < 0.001), overall survival in the two groups was similar. Proportional-hazards regression analysis shows that only resection status is significantly correlated with improved survival (P=0.0068). Within the IPHC group, patients who underwent an R0/R1 resection had increased survival times (11.2 vs. 3.3 months, P=0.015) vs. those who underwent R2 resection. The group who had an R0/R1 resection had 1- and 2-year survival rates of 45% and 45% compared to 16% and 8%, respectively, in the R2 group. Cytoreductive surgery and IPHC is a modality with limited potential for the treatment of peritoneal carcinomatosis from gastric carcinoma. Careful patient selection for this procedure is imperative, and patients in whom an R0/R1 resection can be achieved are the best candidates. PMID- 15120372 TI - Aggressive surgical treatment for T4 gastric cancer. AB - Surgical treatment for locally advanced gastric cancer remains controversial, and many still question the benefits of extended resection. The aim of this study was to evaluate the effectiveness of combined resection of the involved organs with regard to survival in patients with gastric cancer. Between 1993 and 2000, among the 1638 patients with gastric cancer who underwent gastrectomy, 82 were found to have evidence of adjacent organ spread at laparotomy. A retrospective analysis of these patients was performed. Curative resections were carried out in 50 patients, whereas noncurative resections were performed in 32 patients. The 5 year survival rate in the group undergoing curative resection was 36.9%. The survival rate in the R0 group was significantly higher than the survival rate for patients undergoing noncurative resections. There was no significant difference in survival rates between patients with pT3 cancer and those with pT4 cancer. Seventy-one patients were pathologically proved to have lymph node metastasis, and the survival rate for patients with a lymph node ratio greater than 0.2 was lower than that in other groups. In multivariate analysis, peritoneal dissemination, lymph node ratio, and histologic findings were the predictors of survival. Patients with T4 gastric carcinoma, even with lymph node metastasis, might have benefited from aggressive surgery with curative intent. PMID- 15120373 TI - 1990-2001 US general surgery chief resident gastric surgery operative experience: analysis of paradigm shift. AB - The almost complete disappearance of benign gastric ulcer disease has led to the perception that there may be an insufficient gastric surgery experience for surgery residents. This study analyzed resident-reported gastric procedure experience by chief residents from U.S. programs. The Resident Statistic Summaries (Report C) for 1990-2001 were compiled and analyzed. Results are expressed as the average number of operations performed per resident, standard deviation (SD), and the percentage (%) of total gastric operative cases. For all gastric-related surgery, the average reported cases per chief resident ranged from 9.8-12.4 with a peak in 1990 and a nadir in 1999; in 2001 the reported case average was 11.3 (SD ranged from 6-8). Over the same interval, vagotomy decreased from 24% in 1990 to 7% in 2001, whereas gastric-reduction operations increased from 5%-34%. Total gastrectomy remained a constant less than 1.0 per chief resident (range 0.6-0.8), whereas partial gastric resection (PGR) was unchanged. The percentage of all types of gastric resections slightly diminished from 34% in 1990 to 29% in 2001. U.S. surgical chief residents report a widely variable experience in gastric surgery over the period analyzed. However, their overall experience has not significantly diminished since 1990 although specific procedural volume has varied. PMID- 15120374 TI - Prospective evaluation of biliopancreatic diversion with Roux-en-Y gastric bypass in the super obese. AB - The aim of this study was to determine prospectively the efficacy and safety of the biliopancreatic diversion with Roux-en-Y gastric bypass (BPD with RYGBP) procedure used as the primary bariatric procedure in super obese patients. The main characteristics of the BPD with RYGBP procedure were a gastric pouch of 15 +/- 5 ml, biliopancreatic limb of 200 cm, common limb of 100 cm, and alimentary limb of the remainder of the small intestine. From June 1994 through July 2003, 132 super obese patients (body mass index [BMI]: 57 +/- 7), with an incidence of comorbidities 6 +/- 2 per patient, underwent BPD with RYGBP and subsequent follow up. Mean follow-up time was 29 +/- 14 months. Maximum weight loss was achieved at 18 months postoperative with average excess weight loss (EWL) 65%, average initial weight loss (IWL) 39%, and average BMI 35 kg/m(2). Thereafter, a decline was observed with EWL stabilizing at around 50%, IWL at around 30%, and BMI at around 40 kg/m(2), respectively, by the end of the study period. The majority of preexisting comorbidities were permanently resolved by the 6-month follow-up visit. Early mortality was 1% and early morbidity was 11%. Late morbidity was 27%, half of which was due to incisional hernia. Deficiencies of microelements were mild and successfully treated with additional oral supplementation. The incidence of hypoalbuminemia was 3% and there were no hepatic complications. We conclude that BPD with RYGBP is a safe and effective procedure for the super obese with few metabolic complications. PMID- 15120375 TI - Pneumoperitoneum from gas gangrene of the pancreas: three unusual findings in a single case. AB - A 62-year-old man was first seen with acute pancreatitis with diffuse intrapancreatic gas and pneumoperitoneum. An immediate exploratory operation revealed diffuse pancreatic necrosis but no perforated viscus; postoperatively, the patient rapidly died. This case represents a constellation of extremely rare findings: Clostridium perfringens infection of the pancreas, pancreatic emphysema or "gas gangrene," and pneumoperitoneum without a perforated viscus. PMID- 15120376 TI - Laparoscopic distal pancreatectomy combined with preservation of the spleen for cystic neoplasms of the pancreas. AB - The precise role of laparoscopy in the resection of cystic neoplasms of the pancreas (CyNP) remains unknown. In addition, the question of spleen-preserving distal pancreatectomy is controversial. This report evaluates the feasibility and outcome of laparoscopic spleen-preserving distal pancreatectomy (LapSPDP) in 19 patients (17 women and 2 men) with CyNP. A prospective comparison was made between 11 consecutive patients (group I) with splenic vessel preservation (SVP) and 8 patients (group II) without SVP (Warshaw technique). This study used color Doppler ultrasound (CDUS) as a tool to identify patients at high risk for postoperative splenic complications. The mean tumor size was, in both groups, 5 cm. In group I, with an intent-to-treat basis of SVP, only in 54.5% of patients the spleen was preserved with an intact splenic artery and vein; in the remainder, conversion to the Warshaw technique was required for intraoperative bleeding. Evaluation of intraoperative factors showed that the mean operative time was significantly shorter (165 vs. 222 minutes) and the mean blood loss significantly lower (225 vs. 495 mL) in the group of LapSPDP with the Warshaw technique. No patients required blood transfusion in both groups. The overall conversion rate was 0%. The overall rate of pancreatic fistula was 15% and it was classified as biochemical leak (no clinical symptomatology). Overall splenic complications were observed in 16.6% of patients but occurred only in three patients undergoing LapSPDP with the Warshaw technique; CDUS showed in 2 patients a focal splenic infarct; the third patient had an initial hospital stay of 5 days, was readmitted 2 days later for a massive splenic necrosis, and splenectomy was performed. The overall hospital stay was 5.7 days. At mean follow up of 22 months (range 6-42), there have been no local recurrences. PMID- 15120377 TI - A reappraisal of preoperative chemoradiation for localized pancreatic head ductal adenocarcinoma in a 5-year single-institution experience. AB - Resection of localized pancreatic head ductal adenocarcinoma (LPHDA) has a limited impact on survival. Mechanisms of improvement provided by preoperative chemoradiation therapy (CRT) remain under debate. This study analyzes the outcome of patients treated for LPHDA to delineate the benefits of CRT. Among 87 patients with LPHDA, 17 had a pancreaticoduodenectomy alone (group I). Thirty-nine with initially resectable cancers received CRT with 5-fluorouracil-based chemotherapy (group II). Thirty-one with initially unresectable cancers were similarly treated by CRT (group III). Patients in groups II and III were restaged after completion of CRT. In patients with resectable disease, resection was planned. Patients in groups I and II were statistically comparable in terms of age, sex, and pretherapeutic stage. Median survival and 2-year overall survival in group I were 13.7 months and 31%, respectively. In group II, 23 patients (59%) had a pancreaticoduodenectomy (group IIa) and 16 patients (41%) did not have resection (group IIb). Median survival and 2-year overall survival were as follows: group IIa, 26.6 months and 51%; and group IIb, 6.1 months and 0%, respectively. In group IIa, pathologic examination revealed eight major responses (35%) including two sterilized specimens, and none of the patients had locoregional recurrence. In group III, none of the patients had resection, and median survival was 8 months with one 2-year survivor. Patient selection appears to play a major role with regard to results achieved with preoperative CRT followed by pancreaticoduodenectomy. However, a high histologic response rate and excellent local control can also be achieved. PMID- 15120378 TI - COX-2 inhibition results in alterations in nuclear factor (NF)-kappaB activation but not cytokine production in acute pancreatitis. AB - Acute pancreatitis is characterized by local inflammation and cytokine production, and release is thought to contribute to this process. Nuclear factor (NF)-kappaB activation and cytokine production are linked and inhibition of NF kappaB has been shown to decrease the severity of pancreatitis. We have shown that inhibition of COX-2 ameliorates pancreatitis; however, the mechanism by which this effect occurs is unclear. Swiss Webster mice were injected intraperitoneally with either saline (control) or caerulein (CAE; 50 mg/kg) hourly for 8 hours; mice receiving CAE were further subdivided to receive saline or the cyclooxygenase-2 (COX-2) selective inhibitor (SC-58125; 10 mg, intraperitoneally) at the time of the first injection of CAE. Pancreata were harvested, histologic sections were scored, and protein was extracted to determine cytokine (interleukin [IL]-6, IL-1beta) levels and NF-kappaB subunits by ELISA and NF-kappaB activation by gel shift. In addition, serum was collected for measurement of cytokines. COX-2 inhibition resulted in decreased inflammation and a decrease in NF-kappaB activation. IL-6 and IL-1beta levels after COX-2 inhibition, however, remained elevated to levels equivalent to those of mice with histologic inflammation after CAE alone. COX-2 inhibition decreases inflammation as well as late-phase NF-kappaB activation but does not diminish levels of inflammatory cytokines, thus suggesting a two-phase activator of NF-kappaB. The attenuation of inflammation, despite unaltered cytokine levels, suggests that cytokines may not be critical for the inflammatory phase of pancreatitis. PMID- 15120380 TI - A phylogeny of the ITS and ETS for Montanoa (Asteraceae: Heliantheae). AB - A phylogeny of the genus Montanoa based on the internal transcribed spacer (ITS) and the external transcribed spacer (ETS) is presented. Each of the two clades revealed by the Bayesian and parsimony analyses has approximately half of the number of species in the genus. One lineage is composed mostly of central and southern Mexican species whereas the other lineage contains those species endemic to Mesoamerica and South America. The molecular phylogeny is compared to previous phylogenetic hypotheses based on morphological characters. Key features in the structure of the capitulum such as pale morphology, heavily used in the past to construct hypotheses of relationship within the genus, are viewed as of minimal value to circumscribe natural groups. The relationships of Montanoa to other genera in the Heliantheae are briefly discussed. PMID- 15120379 TI - Celiac axis occlusion with replaced common hepatic artery and pancreatoduodenectomy. AB - A rare case of intraductal papillary mucinous tumor of the pancreas associated with a replaced common hepatic artery and celiac axis occlusion, which was treated by pancreatoduodenectomy, is reported. In this patient, the celiac trunk was occluded at its root and the splenic and left gastric artery could be visualized serially via the enlarged collateral artery on superior mesenteric arteriography. At surgery, the collateral artery was carefully preserved and pancreatoduodenectomy was successfully performed without ischemia of the stomach, spleen, and remnant pancreas. Although celiac axis occlusion is an uncommon finding for patients undergoing pancreatoduodenectomy, we recommend performing celio-mesenteric angiography before pancreatoduodenectomy, and, at surgery, clamping of the gastroduodenal artery is required for patients with celiac axis occlusion. PMID- 15120381 TI - Supraordinal phylogenetic relationships of Lecanoromycetes based on a Bayesian analysis of combined nuclear and mitochondrial sequences. AB - Phylogenetic relationships of lichen-forming discomycetes and their relatives in the class Lecanoromycetes were examined by using nuclear large subunit and mitochondrial small subunit ribosomal DNA sequences. Ninety-eight partial sequences of 53 ascomycetes were generated and aligned with the corresponding sequences retrieved from GenBank resulting in an alignment of 100 taxa that was analyzed using a Bayesian approach with Markov chain Monte Carlo (B/MCMC) methods. The analysis revealed the monophyly of the Lecanoromycetes with two major clades: one clade including the monophyletic orders Graphidales and Ostropales and the paraphyletic Gyalectales, the other clade including the monophyletic Lecanorales (incl. Caliciales, Peltigerales, and Teloschistales) and a clade containing the polyphyletic Agyriales, a yet undescribed order Umbilicariales (including Elixiaceae and Umbilicariaceae), and Pertusariales. The monophyly of the Pertusariales was not resolved. Testing of alternative hypotheses revealed that a placement of Chaetothyriomycetes and Eurotiomycetes within Lecanoromycetes and the monophyly of Agyriales s. lat. (incl. Elixiaceae and Schaereriaceae) and Ostropales s. lat. (incl. Graphidales) can be rejected, while monophyly of Gyalectales and the Pertusariales and placement of Umbilicariales on the Lecanorales branch cannot be rejected with the current data set. PMID- 15120383 TI - The phylogenetic history of Selaginellaceae based on DNA sequences from the plastid and nucleus: extreme substitution rates and rate heterogeneity. AB - Molecular phylogenetic research on Selaginellaceae has focused on the plastid gene rbcL, which in this family has unusually high substitution rates. Here we develop a molecular data set from the nuclear 26S ribosomal DNA gene with the aim of evaluating and extending the results of previous phylogenetic research. The 26S rDNA and the rbcL regions were sequenced for a sample of 23 species, which represent the main elements of species diversity in the family. The data were analysed independently and in combination using both maximum parsimony and Bayesian inference. Although several between genome differences were found, the general pattern of relationships uncovered by all analyses was very similar. Results corroborate the previous study supporting new groupings not previously recognised on morphological grounds. Substitution rates in the 26S rDNA were also found to be high (26% informative) for the region analysed, but lower than for rbcL (37% informative). These data indicate that high substitution rates might be widespread in all three genomes (i.e., plastid, mitochondrion, and nucleus). PMID- 15120385 TI - Independence of alignment and tree search. AB - I assert that similarity is the appropriate homology criterion for sequence alignment, as it is with morphology. Methods that select among alignments using parsimony-based tree lengths, as implemented in MALIGN and POY, arrange the data such that they are consistent with a minimum-evolution model. When combining data sets in phylogenetic analyses, we are not trying to reinforce our earlier hypotheses about relationships, but rather to test them. The severity of this test is compromised when congruence with other characters is favored when selecting among alignment parameters. PMID- 15120382 TI - Molecular phylogenetics and biogeography of transisthmian and amphi-Atlantic needlefishes (Belonidae: Strongylura and Tylosurus): perspectives on New World marine speciation. AB - Phylogenetic relationships among New World and eastern Atlantic species in the belonid genera Strongylura and Tylosurus were hypothesized using 3689bp of nucleotide sequence; including the entire mitochondrial (mtDNA) ATP synthase 6 and 8 genes; partial cytochrome b; 12S and 16S ribosomal genes; and introns and exons, 2 and 3 of the nuclear-encoded creatine kinase B gene. Concordant mtDNA and nuclear genealogies permitted well-supported inference of species relationships within Strongylura and Tylosurus, and of the chronology of diversification in the two genera. Our phylogenetic hypothesis permitted an assessment of Rosen's [Syst. Zool. 24 (1975) 431] model of species diversification across the eastern Atlantic to eastern Pacific marine biogeographic track. The spatial predictions of the Rosen model were generally supported, but not the temporal predictions. Furthermore, long branches leading to terminal Belonidae indicated that many species have persisted for millions of years or that nucleotide substitution rates were elevated for some clades. Though heterogeneity of nucleotide substitution rate was indicated across some belonid lineages, molecular clock estimates were used to hypothesize biogeographic scenarios for Strongylura across the eastern Pacific and Atlantic region. Furthermore, use of a molecular clock indicated; that early diversification among contemporary Strongylura may have been initiated by changes in Atlantic Ocean circulation precipitated by closure of the Tethys Sea; and provided approximate dates for the isolation of the freshwater species on the American continents. PMID- 15120384 TI - Efficiencies of maximum likelihood methods of phylogenetic inferences when different substitution models are used. AB - Choice of a substitution model is a crucial step in the maximum likelihood (ML) method of phylogenetic inference, and investigators tend to prefer complex mathematical models to simple ones. However, when complex models with many parameters are used, the extent of noise in statistical inferences increases, and thus complex models may not produce the true topology with a higher probability than simple ones. This problem was studied using computer simulation. When the number of nucleotides used was relatively large (1000 bp), the HKY+Gamma model showed smaller d(T) topological distance between the inferred and the true trees) than the JC and Kimura models. In the cases of shorter sequences (300 bp) simpler model and search algorithm such as JC model and SA+NNI search were found to be as efficient as more complicated searches and models in terms of topological distances, although the topologies obtained under HKY+Gamma model had the highest likelihood values. The performance of relatively simple search algorithm SA+NNI was found to be essentially the same as that of more extensive SA+TBR search under all models studied. Similarly to the conclusions reached by Takahashi and Nei [Mol. Biol. Evol. 17 (2000) 1251], our results indicate that simple models can be as efficient as complex models, and that use of complex models does not necessarily give more reliable trees compared with simple models. PMID- 15120386 TI - Phylogeny of ants (Formicidae) based on morphology and DNA sequence data. AB - In order to reconstruct ants' phylogeny, we analysed DNA sequences for two nuclear genes, abdominal-A and Ultrabithorax, from 49 species of ants and two outgroups. As these genes control the development of the first segments of the abdomen in insects, which are very variable in ants (petiole, postpetiole, and gaster constriction), we hypothesized that the morphological variations between the subfamilies may be correlated with mutations of some abd-A or Ubx regions. Contrarily to our hypothesis, these sequences are highly conserved. The differences observed concern mainly third codon positions and present some saturation. Phylogenetic reconstructions were carried out using the genetic raw sequence data and by combining them with a set of morphological data (Total Evidence). Relations among subfamilies of ants remains poorly resolved with molecular data only, but adding these data to morphological characters confirms and reinforce the topology of : a Poneroid complex [Ponerinae, Cerapachyinae, Leptanillinae and army ants], a Formicoid complex [Dolichoderinae, Formicinae] and a Myrmecoid complex [Myrmicinae, Myrmeciinae, Pseudomyrmecinae, Nothomyrmeciinae]. Our molecular results allow resolution near the branch tips and three subfamilies (Dolichoderinae, Formicinae and Pseudomyrmecinae) always appear as monophyletic. The Formicinae and the Dolichoderinae have close relationships. The Camponotini appear as a strong clade inside the Formicinae. The Ponerinae are separated in two parts: the Ectatommini and all other tribes. The Cerapachyinae, Dorylinae, and Ecitoninae belong to the same clade, the Cerapachyinae being confirmed in their subfamily status. The Myrmicinae appears to be very heterogeneous, with the Attini forming a very stable and well separated group. PMID- 15120387 TI - Phylogeny of Panax using chloroplast trnC-trnD intergenic region and the utility of trnC-trnD in interspecific studies of plants. AB - Sequences of the chloroplast trnC-trnD region and the internal transcribed spacer (ITS) regions of nuclear ribosomal DNA were obtained for all species of Panax L. (the ginseng plant genus, Araliaceae) to reconstruct phylogenetic relationships. The trnC-trnD phylogeny is congruent with the ITS phylogeny for the diploid taxa of Panax. This study is the first use of the trnC-trnD sequence data for phylogenetic analysis at the interspecific level. We evaluated this DNA region for its phylogenetic utility at the lower taxonomic level for flowering plants. The trnC-trnD region includes the trnC-petN intergenic spacer, the petN gene, the petN-psbM intergenic spacer, the psbM gene, and the psbM-trnD intergenic spacer. The petN and psbM genes are small, 90 and 104-114 bp across angiosperms, respectively, and have conserved sequences. We have designed universal amplification and sequencing primers within these two genes. Using these primers, we have successfully amplified the entire trnC-trnD region for a diversity of flowering plant groups, including Aralia L. (Araliaceae), Calycanthus L. (Calycanthaceae), Corylus L. (Betulaceae), Hamamelis L. (Hamamelidaceae), Hydrocotyle L. (Apiaceae), Illigera Blume (Hernandiaceae), Nelumbo Adans. (Nelumbonaceae), Nolana L. ex L.f. (Solanaceae), Prunus L. (Rosaceae), and Staphylea L. (Staphyleaceae). In Panax, the trnC-trnD region provides a similar number of informative phylogenetic characters as the ITS regions and a slightly higher number of informative characters than the chloroplast ndhF gene. We thus demonstrate the utility of the trnC-trnD region for lower-level phylogenetic studies in flowering plants. PMID- 15120388 TI - Molecular phylogeny and possible scenario of ponyfish (Perciformes:Leiognathidae) evolution. AB - The family Leiognathidae, commonly known as ponyfish or slip mouth, comprises three genera, each being characterized mainly by mouth morphology. To date, however, neither the phylogenetic relationships within the family nor monophyly of the genera has been tested. The phylogenetic relationships among 14 species of Leiognathidae, inferred from two protein coding mitochondrial genes (ND4 and 5), indicated monophyly of the studied species form genera Gazza and Secutor, and paraphyly of the genus Leiognathus, with L. equulus occupying a basal branch of the family. The relationships allowed phylogenetic analyses of mouthpart structures and light organ systems. The results suggested that the morphology of the upwardly and forwardly protractile mouth types (latter with canine-like teeth) are phylogenetically informative, and the downwardly protractile mouth type being ancestral in the family. The results also suggested that internal sexual dimorphism of the light organ system was present in the common ancestor of a sister clade to L. equulus, whereas external sexual dimorphism seems to have evolved subsequently in two monophyletic subgroups. PMID- 15120389 TI - Genetic variation in an endemic salamander, Salamandra atra, using amplified fragment length polymorphism. AB - The pattern of genetic differentiation of the endemic alpine salamander, Salamandra atra, has been studied using amplified fragment length polymorphism (AFLP) from 11 populations throughout the range of the two currently recognized subspecies, atra and aurorae. Five different primer combinations produced 706 bands and were analyzed by constructing a phylogenetic tree using NJ and principal component analysis. Significant genetic variation was revealed by AFLP between and within populations but, our results show a lack of genetic structure. AFLP markers seems to be unsuitable to investigate complex and recent diversification. PMID- 15120390 TI - Phylogeny of the Polytrichales (Bryophyta) based on simultaneous analysis of molecular and morphological data. AB - Phylogenetic analyses of Polytrichales were conducted using morphology and sequence data from the chloroplast genes rbcL and rps4 plus the trnL-F gene region, part of the mitochondrial nad5 and the nuclear-encoded 18S rDNA. Our analyses included 46 species representing all genera of Polytrichales. Phylogenetic trees were constructed with simultaneous parsimony analyses of all sequences plus morphology and separate combinations of sequence data only. Results lend support for recognition of Polytrichales as a monophyletic entity. Oedipodium griffithianum appears as a sister taxon to Polytrichales or as a sister taxon of all mosses excluding Sphagnales and Andreaeles. Within Polytrichales, Alophosia and Atrichopsis, species without the adaxial lamellae (in Atrichopsis present but poorly developed on male gametophyte) otherwise typical of the group are sister to the remaining species followed by a clade including Bartramiopsis and Lyellia, species with adaxial lamellae covering only the central portion of the leaves. Six taxa with an exclusively Southern Hemisphere distribution form a grade between the basal lineages and a clade including genera that are mostly confined to the Northern Hemisphere. PMID- 15120391 TI - Range expansion leading to departures from neutrality in the nonsymbiotic hemoglobin gene and the cpDNA trnL-trnF intergenic spacer in Trema dielsiana (Ulmaceae). AB - The population genetics and phylogeography of Trema dielsiana in Taiwan were inferred from genetic diversity at the nonsymbiotic hemoglobin gene and the trnL trnF intergenic spacer of cpDNA. Reduced genetic variation was detected in these two unlinked genes. The gene genealogy of the hemoglobin locus recovered two lineages corresponding to the western and eastern regions of Taiwan. This pattern is compatible with a past fragmentation event revealed by phylogeographical analyses. To distinguish between selective departures from neutrality (i.e., heterogeneous processes) and demographic (homogeneous) processes, Hahn et al.'s heterogeneity test was conducted on the hemoglobin gene. Lack of significant differences in Tajima's D statistics between synonymous and nonsynonymous mutations indicates that homogeneous processes may have played a key role in governing the evolution of the functional locus. Significantly negative Tajima's D estimates for both overall exons and introns of the hemoglobin gene as well as for the cpDNA intergenic spacer support a phylogeographical hypothesis of range expansion after genetic bottlenecks. High level of genetic variation and a negative Tajima's D statistic suggests a possible northern refugium that may have harbored populations during the glacial maximum. PMID- 15120392 TI - Phylogeny and evolution of the Australo-Papuan honeyeaters (Passeriformes, Meliphagidae). AB - We analyzed nucleotide variation at four loci for 75 species to produce a phylogenetic hypothesis for the Meliphagidae, and to examine the evolution and biogeographic history of the Meliphagidae. Both maximum parsimony and Bayesian methods of phylogenetic analysis were employed. The family was found to be monophyletic, though the genera Certhionyx, Anthochaera, and Phylidonyris were not. Four major clades were recovered and the spinebills (Acanthorhynchus) formed the sister clade to the remainder of the family in most analyses. The Australian endemic arid-adapted chats (Epthianura, Ashbyia) were found to be nested deeply within the family Meliphagidae. No evidence was found to support the hypothesis of separate New Guinean and Australian endemic radiations, nor of a close phylogenetic relationship between taxa from the New Guinea highlands and those from Australian northern rainforests. PMID- 15120393 TI - Horned lizard (Phrynosoma) phylogeny inferred from mitochondrial genes and morphological characters: understanding conflicts using multiple approaches. AB - The genus Phrynosoma includes 13 species of North American lizards characterized by unique and highly derived morphologies and ecologies. Understanding interspecific relationships within this genus is essential for testing hypotheses about character evolution in this group. We analyzed mitochondrial ND4 and cytochrome b gene sequence data from all species of Phrynosoma in conjunction with a previously published dataset including 12S and 16S rRNA gene sequences and morphological characters. We used multiple phylogenetic methods and diagnostic tests for data combinability and taxonomic congruence to investigate the data in separate and combined analyses. Separate data partitions resulted in several well supported lineages, but taxonomic congruence was lacking between topologies from separate and combined analyses. Partitioned Bremer support analyses also reveals conflict between data partitions in certain tree regions. When taxa associated with well-supported clades were removed from analyses, phylogenetic signal was lost. Combined, our results initially suggest conflict between data partitions, but further tests show the data are only appropriate for phylogenetic reconstruction of those parts of the topology that were well resolved. Nonetheless, our data analyses reveal five well-supported clades: (1) Phrynosoma ditmarsi and Phrynosoma hernandesi, (2) P. ditmarsi, P. hernandesi, and Phrynosoma douglasii, (3) P. ditmarsi, P. hernandesi, P. douglasii, and Phrynosoma orbiculare, (4) Phrynosoma mcallii and Phrynosoma platyrhinos, and (5) Phrynosoma braconnieri and Phrynosoma taurus. PMID- 15120394 TI - The phylogenetic position of the zokors (Myospalacinae) and comments on the families of muroids (Rodentia). AB - Recent molecular studies have concluded that the genus Myospalax evolved from within the rodent subfamily Cricetinae. This conclusion was tested using the complete sequences from the mitochondrial 12S rRNA and cytochrome b genes. Based on our analyses, Myospalax appears to be sister to a clade containing the subfamilies Spalacinae and Rhizomyinae, and all three of these lineages appear to be basal to the superfamily Muroidea. Based on the position of these three lineages, we suggest that they be placed in a distinct family, the Spalacidae, rather than subsumed as subfamilies in the family Muridae. Finally, our analyses suggest that the earlier placement of Myospalax as a member of the Cricetinae is the result of a single misidentified specimen, which was not a Myospalax. PMID- 15120395 TI - Phylogenetic relationships of glyptosternoid fishes (Siluriformes: Sisoridae) inferred from mitochondrial cytochrome b gene sequences. AB - To explore phylogenetic relationships among glyptosternoid fishes, we determined nucleotide sequences of the complete mitochondrial cytochrome b gene region (1138 base pair). Thirteen species of glyptosternoid fishes and six species of non glyptosternoids represent 10 sisorid genera were examined. Molecular phylogenetic trees were constructed using the maximum parsimony, minimum evolution, maximum likelihood, and Bayesian methods. Bayesian and maximum likelihood analyses support the monophyly of glyptosternoids, but our hypothesis of internal relationships differs from previous hypothesis. Results indicated that glyptosternoid is a monophyletic group and genera Glyptosternum and Exostoma are two basal species having a primitive position among it. Genera Euchiloglanis and Pareuchiloglanis form a sister-group. Then they form a sister-group with Pseudexostoma plus Oreoglanis. Our result also found that Pareuchiloglanis anteanalis might be considered as the synonyms of Parechiloglanis sinensis, and genus Euchiloglanis might have only one valid species, Euchiloglanis davidi. PMID- 15120396 TI - Phylogenetic relationships among Pneumocystis from Asian macaques inferred from mitochondrial rRNA sequences. AB - The presence of Pneumocystis organisms was detected by nested-PCR at mitochondrial large subunit (mtLSU) rRNA gene in 23 respiratory samples from Asian macaques representing two species: Macaca mulatta and M. fascicularis. A very high level of sequence heterogeneity was detected with 18 original sequence types. Two genetic groups of Pneumocystis could be distinguished from the samples. Within each group, the extent of genetic divergence was low (2.5+/-1.4% in group 1 and 2.3+/-1.7% in group 2). Genetic divergences were systematically higher when macaque-derived sequence types were compared with Pneumocystis mtLSU sequences from other primate species (from 5.3+/-2.7% to 19.3+/-3.0%). The two macaque-derived groups may be considered as distinct Pneumocystis species. Surprisingly, these Pneumocystis species were recovered from both M. mulatta and M. fascicularis suggesting that host-species restriction may not systematically occur in the genus Pneumocystis. Alternatively, these observations question about the species concept in macaques. PMID- 15120397 TI - Molecular phylogenetics and taxonomy of the African mole-rats, genus Cryptomys and the new genus Coetomys Gray, 1864. AB - Cryptomys represents the most speciose and widely distributed genus of the Bathyergidae (Mammalia; Rodentia), a family of mole-rats endemic to sub-Saharan Africa. Throughout its range in southern, central, and western Africa, Cryptomys displays diversity in terms of morphology, patterns of behavior, and chromosome number, thus complicating the systematics of the group. A molecular phylogeny was obtained by separate and combined analyses of the mitochondrial 12S rRNA and intron I of the nuclear transthyretin gene for chromosomally and geographically diverse populations of Cryptomys. Our results show that Cryptomys sensu lato is comprised of two distinct and divergent monophyletic clades: hottentotus and mechowi. Based on our analyses, we propose the elevation of the mechowi clade to the genus Coetomys, retaining Cryptomys as the generic epithet of the South African hottentotus clade. This is supported by: (1) reciprocal monophyly of the two lineages based on independent datasets, (2) the level of sequence divergence observed between these lineages relative to other genera (i.e., Bathyergus and Georychus), and (3) different patterns of chromosomal evolution, with Cryptomys sensu stricto being karyotypically conserved (2N=54) and Coetomys exhibiting high karyotypic diversity (2N=40-78). PMID- 15120398 TI - Sciurid phylogeny and the paraphyly of Holarctic ground squirrels (Spermophilus). AB - The squirrel family, Sciuridae, is one of the largest and most widely dispersed families of mammals. In spite of the wide distribution and conspicuousness of this group, phylogenetic relationships remain poorly understood. We used DNA sequence data from the mitochondrial cytochrome b gene of 114 species in 21 genera to infer phylogenetic relationships among sciurids based on maximum parsimony and Bayesian phylogenetic methods. Although we evaluated more complex alternative models of nucleotide substitution to reconstruct Bayesian phylogenies, none provided a better fit to the data than the GTR+G+I model. We used the reconstructed phylogenies to evaluate the current taxonomy of the Sciuridae. At essentially all levels of relationships, we found the phylogeny of squirrels to be in substantial conflict with the current taxonomy. At the highest level, the flying squirrels do not represent a basal divergence, and the current division of Sciuridae into two subfamilies is therefore not phylogenetically informative. At the tribal level, the Neotropical pygmy squirrel, Sciurillus, represents a basal divergence and is not closely related to the other members of the tribe Sciurini. At the genus level, the sciurine genus Sciurus is paraphyletic with respect to the dwarf squirrels (Microsciurus), and the Holarctic ground squirrels (Spermophilus) are paraphyletic with respect to antelope squirrels (Ammospermophilus), prairie dogs (Cynomys), and marmots (Marmota). Finally, several species of chipmunks and Holarctic ground squirrels do not appear monophyletic, indicating a need for reevaluation of alpha taxonomy. PMID- 15120399 TI - Turtle phylogeny: insights from a novel nuclear intron. AB - Introns have gained considerable popularity as markers for molecular phylogenetics. However, no primers exist for a nuclear intron that amplifies across all turtles. Available data from morphology and mitochondrial DNA have not unambiguously resolved relationships within the superfamily Trionychoidea and the family Chelidae, which together form a large portion of extant turtle diversity. We tested the phylogenetic utility of a novel intron from the RNA fingerprint protein 35 (R35) as applied to these two areas of turtle systematics. We found the intron to be a single-copy locus that provides excellent resolving power for lineages among turtles, though problems with alignment made it impossible to infer deeper amniote relationships. Maximum parsimony and maximum likelihood both demonstrated the polyphyly of Trionychoidea and the reciprocal monophyly of Australian/New Guinea and South American chelid turtles. This is the first study to resolve such relationships with strong statistical support, and we suggest that R35 holds great promise for resolving additional persistent problems in the phylogeny of living turtles. PMID- 15120400 TI - Unraveling the phylogeny of polygrammoid ferns (Polypodiaceae and Grammitidaceae): exploring aspects of the diversification of epiphytic plants. AB - We explore the phylogeny of the polygrammoid ferns using nucleotide sequences derived from three plastid loci for each of 98 selected species. Our analyses recovered four major monophyletic lineages: the loxogrammoids, two clades consisting of taxa restricted to the Old World, and a largely neotropical clade that also includes the pantropical Grammitidaceae. The loxogrammoid lineage diverges first and is sister to a large clade comprising the three remaining species-rich lineages. One paleotropical clade includes the drynarioid and selligueoid ferns, whereas the second paleotropical clade includes the platycerioids, lepisoroids, microsoroids, and their relatives. The grammitids nest within the neotropical clade, although the sister taxon of this circum tropic, epiphytic group remains ambiguous. Microsorum and Polypodium, as traditionally defined, were recovered as polyphyletic. The relatively short branch lengths of the deepest clades contrast with the long branch lengths leading to the terminal groups. This suggests that the polygrammoid ferns arose through an old, rapid radiation. Our analysis also reveals that the rate of substitution in the grammitids is remarkably higher relative to other polygrammoids. Disparities in substitution rate may be correlated with one or more features characterizing grammitids, including species richness, chlorophyllous spores, and an extended gametophytic phase. PMID- 15120401 TI - Mitochondrial DNA phylogeography of red deer (Cervus elaphus). AB - In order to understand the origin, phylogeny, and phylogeography of the species Cervus elaphus, we examined the DNA sequence variation of the mitochondrial cytochrome b gene of 51 populations of deer from the entire distribution area of Cervinae with an emphasis on Europe and Asia. Several methods, including maximum parsimony, maximum likelihood, and nested clade analysis, revealed that red deer originated from the area between Kyrgyzstan and Northern India. We found two distinct groups of red deer: a western group consisting of four subgroups and an eastern group consisting of three subgroups. Our mtDNA data do not support the traditional classification of red deer as only one species nor its division into numerous subspecies. The discrepancies between the geographical pattern of differentiation based on mtDNA cytochrome b and the existing specific and subspecific taxonomy based on morphology are discussed. PMID- 15120402 TI - Phylogenetic relationships of Iberian Aphodiini (Coleoptera: Scarabaeidae) based on morphological and molecular data. AB - A phylogeny of Iberian Aphodiini dung beetles was reconstructed based on morphological and molecular data. The data set included a total of 84 variable characters from wing venation, mouthparts, genitalia, and external morphology, as well as mitochondrial partial cytochrome c oxidase I (COI), complete tRNA-Leu (UUR), and partial cytochrome c oxidase II (COII) gene nucleotide sequences (1210 positions). Phylogenetic trees based on molecular data were relatively more resolved than those based on morphological characters. The Bayesian analysis of combined molecular and morphological data provided resolution not achieved by each data set separately. Ammoecius and Aphodius are the first lineages that branch off from the tree, followed by Acrossus, Nimbus, and Heptaulacus. The remaining studied taxa are recovered in a more derived clade that lacks internal resolution. Reconstructed trees based on molecular data showed relatively short internal nodes that were weakly supported. Such pattern may reflect a rapid radiation at the origin of the tribe Aphodiini, but also saturation of mutational changes. Several tests were conducted to discern between both competing hypotheses, as well as to assess the effect of incomplete taxon sampling. PMID- 15120403 TI - Convergence among cave catfishes: long-branch attraction and a Bayesian relative rates test. AB - Convergence has long been of interest to evolutionary biologists. Cave organisms appear to be ideal candidates for studying convergence in morphological, physiological, and developmental traits. Here we report apparent convergence in two cave-catfishes that were described on morphological grounds as congeners: Prietella phreatophila and Prietella lundbergi. We collected mitochondrial DNA sequence data from 10 species of catfishes, representing five of the seven genera in Ictaluridae, as well as seven species from a broad range of siluriform outgroups. Analysis of the sequence data under parsimony supports a monophyletic Prietella. However, both maximum-likelihood and Bayesian analyses support polyphyly of the genus, with P. lundbergi sister to Ictalurus and P. phreatophila sister to Ameiurus. The topological difference between parsimony and the other methods appears to result from long-branch attraction between the Prietella species. Similarly, the sequence data do not support several other relationships within Ictaluridae supported by morphology. We develop a new Bayesian method for examining variation in molecular rates of evolution across a phylogeny. PMID- 15120404 TI - Mitochondrial phylogeography of the long-eared bats (Plecotus) in the Mediterranean Palaearctic and Atlantic Islands. AB - Long-eared bats of the genus Plecotus are widespread and common over most of the western Palaearctic. Based on recent molecular evidence, they proved to represent a complex of several cryptic species, with three new species being described from Europe in 2002. Evolutionary relationships among the different lineages are still fragmentary because of the limited geographic coverage of previous studies. Here we analyze Plecotus mitochondrial DNA sequences from the entire Mediterranean region and Atlantic Islands. Phylogenetic reconstructions group these western Palaearctic Plecotus into two major clades which split at least 5 Myr ago and that are each subdivided into further subgroups. An 'auritus group' includes the traditional P. auritus species and its sister taxon P. macrobullaris (=P. alpinus) plus related specimens from the Middle East. P. auritus and P. macrobullaris have broadly overlapping distributions in Europe, although the latter is apparently more restricted to mountain ranges. The other major clade, the 'austriacus group,' includes the European species P. austriacus and at least two other related taxa from North Africa (including P. teneriffae from the Canary Islands), the Balkans and Anatolia (P. kolombatovici). The sister species of this 'austriacus group' is P. balensis, an Ethiopian endemic. Phylogenetic reconstructions further suggest that P. austriacus reached Madeira during its relatively recent westward expansion through Europe, while the Canary Islands were colonized by a North African ancestor. Although colonization of the two groups of Atlantic Islands by Plecotus bats followed very distinct routes, neither involved lineages from the 'auritus group.' Furthermore, the Strait of Gibraltar perfectly segregates the distinct lineages, which confirms its key role as a geographic barrier. This study also stresses the biogeographical importance of the Mediterranean region, and particularly of North Africa, in understanding the evolution of the western Palaearctic biotas. PMID- 15120405 TI - The black widow spider genus Latrodectus (Araneae: Theridiidae): phylogeny, biogeography, and invasion history. AB - The spider genus Latrodectus includes the widely known black widows, notorious because of the extreme potency of their neurotoxic venom. The genus has a worldwide distribution and comprises 30 currently recognized species, the phylogenetic relationships of which were previously unknown. Several members of the genus are synanthropic, and are increasingly being detected in new localities, an occurrence attributed to human mediated movement. In particular, the nearly cosmopolitan range of the brown widow, Latrodectus geometricus, is a suspected consequence of human transport. Although the taxonomy of the genus has been examined repeatedly, the recognition of taxa within Latrodectus has long been considered problematic due to the difficulty associated with identifying morphological features exhibiting discrete geographic boundaries. This paper presents, to our knowledge, the first phylogenetic hypothesis for the Latrodectus genus and is generated from DNA sequences of the mitochondrial gene cytochrome c oxidase subunit I. We recover two well-supported reciprocally monophyletic clades within the genus: (1) the geometricus clade, consisting of Latrodectus rhodesiensis from Africa, and its is sister species, the cosmopolitan L. geometricus, and (2) the mactans clade containing all other Latrodectus species sampled, including taxa occurring in Africa, the Middle East, Iberian Peninsula, Australia, New Zealand, and North and South America. Recovery of the geometricus and mactans clades is consistent with previous designations of species groups within the genus based on female genitalic morphology. All L. geometricus sampled, consisting of specimens from Africa, Argentina, North America, and Hawaii, were recovered as a strongly supported monophyletic group with minimal amounts of genetic divergence, corroborating the hypothesis that human transport has recently expanded the range of this species. PMID- 15120406 TI - Phylogeography of Varicorhinus barbatulus (Cyprinidae) in Taiwan based on nucleotide variation of mtDNA and allozymes. AB - The phylogeographical patterns and population genetic structures of Varicorhinus barbatulus in Taiwan were investigated based on genetic diversity of 34 allozyme loci and nucleotide sequences of 3' end of the cytochrome b gene, tRNA genes, D loop control region, and the 5' end of the 12S rRNA of mtDNA. Allozyme and mtDNA analyses revealing evident geographical structuring suggest limited gene flow between populations (F(ST)=0.511 and 0.791, respectively). Low genetic variability within populations (P=5.56%; He=0.018) based on allozymes and significantly negative Tajima's D statistics based on mtDNA suggest that most populations in Taiwan may have originated from a small number of founders followed by demographic expansion. The gene genealogy of mtDNA identified six lineages corresponding to major drainages that were separated by the geological barriers due to vicariant events. A minimum spanning network based on nucleotide substitutions reflects divergence from populations of the Miao-li Plateau to northern and southern regions of the island. In contrast to a previous hypothesis that suggests an early invasion to eastern part of Taiwan prior to the lifting of central mountain range some one million years ago, the mtDNA genealogy and molecular dating reveal very recent colonization of the eastern population. Nested clade analyses revealing significant associations between genetic structure and geographical division identify past fragmentation and range expansion as major phylogeographical events that shaped the geographical distribution of this species in Taiwan. PMID- 15120407 TI - Utility of plastid psaB gene sequences for investigating intrafamilial relationships within Orchidaceae. AB - DNA sequences of the plastid gene psaB were completed for 182 species of Orchidaceae (representing 150 different genera) and outgroup families in Asparagales. These data were analyzed using parsimony, and resulting trees were compared to a rbcL phylogeny of Orchidaceae for the same set of taxa after an additional 30 new rbcL sequences were added to a previously published matrix. The psaB tree topology is similar to the rbcL tree, although the psaB data contain less homoplasy and provide greater bootstrap support than rbcL alone. In combination, the two-gene tree recovers the five monophyletic subfamilial clades currently recognized in Orchidaceae, but fails to resolve the positions of Cypripedioideae and Vanilloideae. These new topologies help to clarify some of the anomalous results recovered when rbcL is analyzed alone. Both genes appear to be absent from the plastid genome of several achlorophyllous orchids, but are present in the form of presumably non-functional pseudogenes in Cyrtosia. This study is the first to document the utility of psaB sequences for phylogenetic studies of plants below the family level. PMID- 15120408 TI - Tempo of speciation in a butterfly genus from the Southeast Asian tropics, inferred from mitochondrial and nuclear DNA sequence data. AB - Molecular systematics is frequently beset with phylogenetic results that are not fully resolved. Researchers either state that the absence of resolution is due to character conflict, explosive speciation, or some combination of the two, but seldom do they carefully examine their data to distinguish between these causes. In this study, we exhaustively analyze a set of nuclear and mitochondrial nucleotide data for the Asian tropical butterfly genus Arhopala so as to highlight the causes of polytomies in the phylogenetic trees, and, as a result, to infer important biological events in the history of this genus. We began by using non-parametric statistical methods to determine whether the ambiguously resolved regions in these trees represent hard or soft polytomies. In addition we determined how this correlated to number of inferred changes on branches, using parametric maximum likelihood estimations. Based on congruent patterns in both mitochondrial and nuclear DNA sequences, we concluded that at two stages in the history of Arhopala there have been accelerated instances of speciation. One event, at the base of the phylogeny, generated many of the groups and subgroups currently recognized in this genus, while a later event generated another major clade consisting of both Oriental and Papuan species groups. Based on comparisons of closely related taxa, the ratio of instantaneous rate of evolution between mitochondrial and nuclear DNA evolution is established at approximately 3:1. The earliest radiation is dated between 7 and 11 Ma by a molecular clock analysis, setting the events generating much of the diversity of Arhopala at well before the Pleistocene. Periodical flooding of the Sunda plateau during interglacial periods was, therefore, not responsible for generating the major divisions in the genus Arhopala. Instead, we hypothesize that large-scale climatic changes taking place in the Miocene have induced the early acceleration in speciation. PMID- 15120409 TI - Phylogeny of the Bangia flora of New Zealand suggests a southern origin for Porphyra and Bangia (Bangiales, Rhodophyta). AB - Analysis of nuclear small subunit ribosomal RNA gene (18S rDNA) sequence data from 123 samples of the red algal genus Bangia from mainland New Zealand has revealed diversity exceeding that reported for the genus from any other region in the world. Our study resolves two New Zealand Bangia taxa basal to the order Bangiales, and five clades of Bangia, four of which include New Zealand members. The basal taxa are separated from each other by 139 bp and differ from all other Bangia taxa in the New Zealand region by 103-163 bp over approximately 1750 bp 18S rDNA sequence data. Our results reveal a Bangia flora of previously unsuspected richness, and show that the simple morphology of these organisms obscures significant levels of genetic diversity. The presence of high diversity and retention of basal taxa in New Zealand Bangia raises the prospect that the southern hemisphere, and particularly eastern Gondwana, is not only a centre of diversity, but a centre of origin for the modern Bangiales. PMID- 15120410 TI - Isolation of Hox and Parahox genes in the hemichordate Ptychodera flava and the evolution of deuterostome Hox genes. AB - Because of their importance for proper development of the bilaterian embryo, Hox genes have taken center stage for investigations into the evolution of bilaterian metazoans. Taxonomic surveys of major protostome taxa have shown that Hox genes are also excellent phylogenetic markers, as specific Hox genes are restricted to one of the two great protostome clades, the Lophotrochozoa or the Ecdysozoa, and thus support the phylogenetic relationships as originally deduced by 18S rDNA studies. Deuterostomes are the third major group of bilaterians and consist of three major phyla, the echinoderms, the hemichordates, and the chordates. Most morphological studies have supported Hemichordata+Chordata, whereas molecular studies support Echinodermata+Hemichordata, a clade known as Ambulacraria. To test these competing hypotheses, complete or near complete cDNAs of eight Hox genes and four Parahox genes were isolated from the enteropneust hemichordate Ptychodera flava. Only one copy of each Hox gene was isolated suggesting that the Hox genes of P. flava are arranged in a single cluster. Of particular importance is the isolation of three posterior or Abd-B Hox genes; these genes are only shared with echinoderms, and thus support the monophyly of Ambulacraria. PMID- 15120411 TI - Abstracts of the XIIIth International Vascular Biology Meeting 2004. Toronto, Canada, June 1-5, 2004. PMID- 15120412 TI - A wake-up call to advance women's health. PMID- 15120413 TI - The development and validation of the primary care satisfaction survey for women. AB - Assessing patient satisfaction with health care is becoming an integral component of quality monitoring in health care systems, but existing tools typically were developed to minimize differences related to gender. This paper reports the development and psychometric properties of a new survey instrument to measure women's satisfaction with their primary care. A multisite, cross-sectional validation survey of 1,202 women receiving care in primary care settings in Michigan, North Carolina, and Pennsylvania was conducted. Item response theory (IRT) and factor analysis methods were used to identify three scales in the Primary Care Satisfaction Survey for Women (PCSSW): Communication, Administration and Office Procedures, and Care Coordination and Comprehensiveness. Internal consistency reliability is reported, as well as convergent validity in relation to two generic measures (Medical Outcomes Study [MOS] Visit Satisfaction and Consumer Assessment of Health Plans Survey [CAHPS] overall quality rating); discriminant validity in relation to groups expected to have differing satisfaction levels based on previous literature; predictive validity in relation to behavioral intentions; and explanatory power in overall satisfaction ratings. The 24-item PCSSW may be self-administered or conducted by telephone and may be used in studies to evaluate or improve the quality of primary care for women. PMID- 15120414 TI - Context of care and contraceptive method use. AB - OBJECTIVES: We assessed the relationship between context of care (incorporates insurance status with clinical setting) on contraceptive use among a national sample of reproductive-aged women. Our hypothesis is that compared to privately insured women who receive their health care in private doctors' offices, women who are publicly insured or self-pay and/or receive their health care in a clinic are more likely to use permanent or long-acting contraceptive methods. METHODS: The study population, consisting of 4,358 women surveyed as part of the 1995 National Survey of Family Growth (NSFG) who were both at risk of unintended pregnancy and currently sexually active, was analyzed using polytomous logistic regression. RESULTS: Following adjustment for age, race/ethnicity, marital status, education, income, parity and smoking, there was a trend toward long acting contraceptive use among women with public insurance or who were self-pay, regardless of clinical setting compared to privately insured women seen in private clinics. Self-pay and publicly insured women of low parity tended to use long-acting contraception, as did privately insured women seen in clinics. CONCLUSIONS: Insurance information, as well as clinical setting, may guide clinicians' contraceptive decision-making. PMID- 15120415 TI - Comparing breast cancer risk between lesbians and their heterosexual sisters. AB - PURPOSE: The purpose of this study was to explore the similarities and differences between lesbians and their heterosexual sisters in the established risks for developing breast cancer. METHODS: The design for this study was a matched (lesbian with heterosexual sister) cross-sectional, mail-back, anonymous survey. We distributed the surveys throughout the state of California to English speaking women who identified themselves as lesbians, age 40 and older, and their sisters. Using the modified Gail Breast Cancer Risk model as well as other well established factors associated with the development of breast cancer, we compared the breast cancer risk potential for 324 sister pairs (N = 648). Data were analyzed using paired t-tests, analysis of covariance (ANCOVA), McNemar's chi(2), or the Bowker statistic, as appropriate for the level of data. MAIN FINDINGS: The lesbians had significantly higher 5-year (p <.0001) and lifetime (p =.001) risk for developing breast cancer. The reasons for lesbians' predicted rate of breast cancer were most likely their higher scores on all pregnancy-related variables and the relatively high number of breast biopsies they reported. The lesbians had used birth control pills less (p <. 0001), had significantly fewer pregnancies (p <.0001), children (p <.0001), abortions (p <.0001), and miscarriages (p <.0001) as well as significantly more breast biopsies (p =.02) than did their heterosexual sisters. CONCLUSIONS: A lesbian who comes out to her clinician is relying on the clinician to be informed and be open to discuss her life. When a lesbian has a lump or a suspicious mammogram, she needs her clinician to advocate for her within the health care system because she is at higher risk for having cancer than a heterosexual woman. PMID- 15120416 TI - Estrogenic effects of resveratrol in breast cancer cells expressing mutant and wild-type estrogen receptors: role of AF-1 and AF-2. AB - Resveratrol, a hydroxystilbene found in grapes and wine, has previously been shown to be a non-flavonoid phytoestrogen, and to act as an estrogen receptor (ER) superagonist in MCF-7 cells transiently transfected with estrogen-responsive reporter constructs. Several additional hydroxystilbenes, including diethylstilbestrol (DES) and piceatannol, were tested, and all showed ER agonism or partial agonism, but superagonism was specific to resveratrol. Moreover, superagonism was observed in cells carrying a stably integrated reporter gene, indicating that this phenomenon is not a result of transient transfection. To examine the role of the transcriptional activation function (AF) domains of ERalpha in resveratrol agonism, we compared the effects of resveratrol and estradiol (E2) on expression of exogenous reporter genes and an endogenous estrogen-regulated gene (TGFalpha) in MDA-MB-231 cells stably transfected with wild-type (wt) ERalpha or mutants with deleted or mutated AF domains. In reporter gene assays, cells expressing wtERalpha showed a superagonistic response to resveratrol. Deletion of AF-1 or mutation of AF-2 attenuated the effect of resveratrol disproportionately compared to that of E2, while deletion of AF-2 abrogated the response to both ligands. In TGFalpha expression assays, resveratrol acted as a full agonist in cells expressing wtERalpha. Deletion of AF 1 attenuated stimulation by E2 more severely than that by resveratrol, as did deletion of AF-2. In contrast, mutation of AF-2 left both ligands with a limited ability to induced TGFalpha expression. In summary, the effect of modifying or deleting AF domains depends strongly on the ligand and the target gene. PMID- 15120417 TI - Suppression of human cytochrome P450 aromatase activity by monoclonal and recombinant antibody fragments and identification of a stable antigenic complex. AB - Human cytochrome P450 aromatase (P450arom) is responsible for biosynthesis of estrogens from androgens. Monoclonal antibody MAb3-2C2 to P450arom specifically binds to a conformational epitope and suppresses the enzyme activity in a dose dependent manner. The crystal structure of the Fab fragment of MAb3-2C2 has been used to engineer a recombinant single chain antibody fragment (scFv) and a homodimeric variable domain of the light chain (VL(2)). These recombinant antibody fragments have been expressed in Escherichia coli and purified. Here, we show that the recombinant scFv suppresses P450arom activity with an IC(50) value similar to that of natural MAb3-2C2 F(ab')(2). The recombinant VL(2) also exhibits dose-dependent suppression of the P450arom activity, but at a reduced level, demonstrating that the homodimer is unable to fully mimic the complementarity determining region (CDR) of a variable heavy chain (VH)-VL heterodimer. We prepare and purify a stable complex of P450arom with MAb3-2C2 F(ab')(2) and show that the complex migrates and precipitates as a single molecular assembly. Efforts to crystallize P450arom for structure-function studies have yielded small single crystals. Our results suggest that formation of stable complexes with fragments of the monoclonal antibody could provide an alternative method for crystallization of P450arom. PMID- 15120418 TI - Immediate early gene X-1 (IEX-1), a hydroxytamoxifen regulated gene with increased stimulation in MCF-7 derived resistant breast cancer cells. AB - The efficacy of tamoxifen in breast cancer treatment only lasts a few years and the tumor eventually recurs. We performed selective subtractive hybridization to isolate mRNAs that were differentially expressed in MCF-7 derived cells, in which resistance had been induced through long-term culture in the presence of hydroxytamoxifen (OHT). Among the 15 mRNAs found to be overexpressed, we focused on Immediate early gene X-1 (IEX-1) mRNA because of the recognized contribution of its expression to apoptosis or cell cycle progression, depending on the cell type and culture conditions. We observed that IEX-1 expression was stimulated by OHT, that the degree of increase was greater in resistant cells (four-fold versus 1.5-fold) and that this OHT regulation was estrogen receptor dependent. A detailed study of the IEX-1 promoter indicated that it involved NF-kappaB. Our cells were not cross-resistant to faslodex, a pure antiestrogen, which moreover was inefficient in regulating IEX-1 expression. Altogether, our data suggest that the greater IEX-1 expression in OHT resistant cells is related to their ability to grow in the presence of OHT. Knowledge on the capacity of OHT to stimulate gene expression and its NF-kappaB dependence should contribute to a better understanding of tamoxifen pharmacology and allow new drug strategies to be designed that would delay antiestrogen resistance acquisition. PMID- 15120419 TI - Dehydroepiandrosterone (DHEA) and its sulfate (DHEAS) inhibit the apoptosis in human peripheral blood lymphocytes. AB - BACKGROUND: Dehydroepiandrosterone (DHEA) and DHEA-sulfate (DHEAS) are the major steroid hormones secreted by the adrenal gland. Administration of DHEA has been reported to have beneficial effects on aging, diabetes, and atherosclerosis. Apoptosis is a normal physiologic process that occurs during embryonic development as well as in the maintenance of tissue homeostasis. In this study, we examined the suppressive effect of DHEA(S) on staurosporine-induced apoptosis in human peripheral blood lymphocytes (PBL). METHODS: Apoptosis was induced in human PBL with staurosporine and measured by flow cytometry utilizing Annexin V and propidium iodide (PI) staining. The quantity of FITC+/PI- cells corresponded to early apoptosis, while that of FITC+/PI+ cells corresponded to late apoptosis or secondary necrosis. RESULTS: The fraction of staurosporine-induced early apoptosis but not that of secondary necrosis in PBL was reduced by the treatment with either DHEA or DHEAS. Furthermore, this apoptosis was neither associated with androgen receptor (AR) nor with estrogen receptor (ER). CONCLUSIONS: This is the first study showing that DHEA(S) inhibits apoptosis in human PBL through a mechanism independent of either ARs or ERs. DHEA(S) may be a promising chemopreventive drug for aging, diabetes, and atherosclerosis. PMID- 15120420 TI - Concentration-dependent mitogenic and antiproliferative actions of 2 methoxyestradiol in estrogen receptor-positive human breast cancer cells. AB - We compared in this study the effects of 2-methoxyestradiol (2-MeO-E(2)) on the growth of two estrogen receptor (ER)-negative human breast cancer cell lines (MDA MB-231 and MDA-MB-435s) and two ER-positive human breast cancer cell lines (MCF-7 and T-47D). 2-MeO-E(2) exerted a concentration-dependent antiproliferative action in the ER-negative MDA-MB-231 and MDA-MB-435s cells. The presence or absence of exogenous 17beta-estradiol (E(2)) in the culture medium did not affect the potency and efficacy of 2-MeO-E(2)'s antiproliferative action in these ER negative cells. When the ER-positive MCF-7 and T-47D cells were cultured in a medium supplemented with 10nM of exogenous E(2), 2-MeO-E(2) at 750 nM to 2 microM concentrations exerted a similar antiproliferative effect. However, when the ER positive cell lines were cultured in the absence of exogenous E(2), 2-MeO-E(2) at relatively low concentrations (10-750 nM) had a moderate mitogenic effect, with its apparent efficacy 75-80% of that of E(2). This mitogenic effect of 2-MeO-E(2) was ER-mediated and largely attributable to 2-MeO-E(2)'s residual estrogenic activity on the basis of our following findings: (i) its effect was only manifested in the ER-positive cells but not in the ER-negative cells; (ii) its effect in the ER-positive cells was partially or fully abolished when exogenous E(2) was concomitantly present in the culture medium; (iii) 2-MeO-E(2) retained 1 2% of E(2)'s binding affinity for the human ERalpha and ERbeta, and its mitogenic effect was inhibited in a concentration-dependent manner by ICI-182,780, a pure ER antagonist; and (iv) its effect was not due to its metabolic conversion to 2 hydroxyestradiol. Our timely findings are of importance to the on-going clinical trials designed to evaluate 2-MeO-E(2)'s effectiveness for the treatment of different types (ER-positive or ER-negative) of human breast cancer. This knowledge will improve the design of clinical trials as well as the interpretation of clinical outcomes when 2-MeO-E(2) is used as a single agent therapy or as part of a combination therapy for human breast cancer. PMID- 15120421 TI - In vitro antiprogestational/antiglucocorticoid activity and progestin and glucocorticoid receptor binding of the putative metabolites and synthetic derivatives of CDB-2914, CDB-4124, and mifepristone. AB - In determining the biological profiles of various antiprogestins, it is important to assess the hormonal and antihormonal activity, selectivity, and potency of their proximal metabolites. The early metabolism of mifepristone is characterized by rapid demethylation and hydroxylation. Similar initial metabolic pathways have been proposed for CDB-2914 (CDB: Contraceptive Development Branch of NICHD) and CDB-4124, and their putative metabolites have been synthesized. We have examined the functional activities and potencies, in various cell-based assays, and relative binding affinities (RBAs) for progesterone receptors (PR) and glucocorticoid receptors (GR) of the putative mono- and didemethylated metabolites of CDB-2914, CDB-4124, and mifepristone and of the 17alpha-hydroxy and aromatic A-ring derivatives of CDB-2914 and CDB-4124. The binding affinities of the monodemethylated metabolites for rabbit uterine PR and human PR-A and PR-B were similar to those of the parent compounds. Monodemethylated mifepristone bound to rabbit thymic GR with higher affinity than monodemethylated CDB-2914 or CDB-4124. T47D-CO cells were used to assess inhibition of R5020-stimulated endogenous alkaline phosphatase activity and transactivation of the PRE(2) thymidine kinase (tk)-luciferase (LUC) reporter plasmid in transient transfections. The antiprogestational potency was as follows: mifepristone/CDB 2914/CDB-4124/monodemethylated metabolites (IC(50)'s approximately 10(-9)M) > aromatic A-ring derivatives (IC(50)'s approximately 10(-8)M) > didemethylated/17alpha-hydroxy derivatives (IC(50)'s approximately 10(-7)M). Antiglucocorticoid activity was determined by inhibition of dexamethasone stimulated transcriptional activity in HepG2 cells. The mono- and didemethylated metabolites of CDB-2914 and CDB-4124 had less antiglucocorticoid activity (IC(50)'s approximately 10(-6)M) than monodemethylated mifepristone (IC(50) approximately 10(-8)M) or the other test compounds. At 10(-6)M in transcription assays, none of these compounds showed progestin agonist activity, whereas mifepristone and its monodemethylated metabolite manifested slight glucocorticoid agonist activity. The reduced antiglucocorticoid activity of monodemethylated CDB 2914 and CDB-4124 was confirmed in vivo by the thymus involution assay in adrenalectomized male rats. The aromatic A-ring derivatives-stimulated transcription of an estrogen-responsive reporter plasmid in MCF-7 and T47D-CO human breast cancer cells but were much less potent than estradiol. Taken together, these data suggest that the proximal metabolites of mifepristone, CDB 2914, and CDB-4124 contribute significantly to the antiprogestational activity of the parent compounds in vivo. Furthermore, the reduced antiglucocorticoid activity of CDB-2914 and CDB-4124 compared to mifepristone in vivo may be due in part to decreased activity of their putative proximal metabolites. PMID- 15120422 TI - Vitamin D3 polyunsaturated side-chain analogues (EB1089, GS1590) and the 20-epi vitamin D3 analogue CB1393 suppress parathyroid hormone secretion and mRNA level in bovine parathyroid cells. AB - Several vitamin D analogues, with reduced hypercalcemic and hyperphosphatemic toxicity at therapeutic dosages, are in clinical use for prevention and treatment of secondary hyperparathyroidism (HPT) in chronic renal failure. We have performed a first in vitro evaluation of five vitamin D analogues displaying less calcemic activity in normal rats, considerably more antiproliferative ability and higher transcription activation potential than 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)), with the future prospects in mind to identify even more effective and less calcemic vitamin D analogues for treatment of HPT. The vitamin D analogues EB1089 and GS1590 have polyunsaturated side-chains, whereas HEP187, MC1598 and CB1393 display altered stereochemistry at carbon 20. In primary cultures of bovine parathyroid cells EB1089, GS1590, CB1393 and MC1598 as well as the comparative controls 1,25(OH)(2)D(3), 22-oxacalcitriol (OCT, maxacalcitol), 19-nor-1,25(OH)(2)D(2) (paricalcitol) and 1alpha(OH)D(2) (doxercalciferol) significantly suppressed PTH secretion or reduced PTH mRNA level at 10(-8), 10( 10), and 10(-11)M for all compounds except for MC1598 at the lowest concentration. The analogue HEP187 displayed no PTH suppressive activity. We conclude that the vitamin D analogues EB1089, GS1590 and CB1393 may be suitable for treatment of hyperparathyroidism secondary to uremia and that further evaluation in vivo should be considered. PMID- 15120423 TI - Developmental and hormonal regulated gene expression of fibroblast growth factor 2 (FGF-2) and its receptors in porcine endometrium. AB - This study examined the mRNA levels of the fibroblast growth factor 2 (FGF-2) and two of its receptors, FGFR1IIIc and FGFR2IIIc, at days 12 and 20 of the ovarian cycle (DC 12 and DC 20), days 1 and 12 of pregnancy (DP 1 and DP 12) as well as the influence of progesterone (P) and estradiolbenzoate (EB) on their expression in the endometrium of ovariectomized (ovx) gilts by real-time PCR. Proteins of FGF-2 and FGFR1 were immunolocalized. FGF-2 and FGFR2IIIc mRNAs were always found with a 5- to 30-fold higher absolute concentration compared to FGFR1IIIc. The latter transcript significantly declined between DP 1 and DP 12, whereas FGF-2 and FGFR2IIIc showed no significant changes at that time. FGF-2 transcription was greater at DC 20 than at DC 12, but significantly most transcripts were found in ovx gilts. EB induced a significant suppression of FGF-2 mRNA, an effect which was antagonized by P and even prevented by P+EB. FGFR1IIIc mRNA was significantly increased at DC 20, that of FGFR2IIIc at DC 12 displaying a 10 times higher absolute mRNA amount. Suppression of FGFR1IIIc mRNA by P was abolished by EB while P+EB attenuated this effect. FGFR2IIIc transcripts were equally restrained by P or EB while a combination of both slightly reduced such declines. Localization of FGF-2 and FGFR1 proteins in stromal, glandular and vascular compartments was effected by sex steroids. Both proteins were strongly expressed at DP 12 but not at DP 1. Summarized, differential temporal and spatial localization of FGF-2 and FGFR1 after response to sex steroids support a complex regulation of this ligand receptor system important for proliferation and differentiation of uterine cells including angiogenic processes. While FGFR1IIIc is presumed to be promoted by estradiol FGFR2IIIc appears to be dominated by progesterone implicating different biological importance for a functional endometrium. PMID- 15120424 TI - Sodium chloride regulation of the alpha epithelial amiloride-sensitive sodium channel (alphaENaC) gene requires syntheses of new protein(s). AB - The epithelial amiloride-sensitive sodium channel (ENaC) plays a central role in sodium homeostasis and blood pressure control. The molecular effect of high sodium intake on the ENaC gene is not well known. This study examined the effects of high salt (HS) intake on alphaENaC gene transcription in rat kidney. Rats were injected intraperitoneally with hypertonic (1.5M NaCl) or normal saline solution (three rats per group). The serum sodium concentration of rats injected with hypertonic saline increased significantly 30 min after injection (158 +/- 2 mM versus 140 +/- 1 mM for normal saline injected rats and 139 +/- 1 mM for uninjected rats). At 3 h after injection, serum sodium decreased (144 +/- 1 mM) but remained above the control values (139 +/- 1 mM for normal saline injected rats, 139 +/- 1 mM for uninjected rats). The serum aldosterone decreased 1.5 and 3 h after the hypertonic saline injection (217 +/- 10 and 139 +/- 23 pg/ml for hypertonic saline injected rats, 358 +/- 2 pg/ml for uninjected rats). The kidney cortex was dissected macroscopically and total RNA was isolated at 1.5 and 3 h after treatment. Semi-quantitative RT-PCR studies revealed that following hypertonic saline treatment, alphaENaC mRNA levels were dramatically downregulated, compared with controls, as early as 1.5h. Western blot analysis showed similar patterns of protein downregulation. Inhibition of protein synthesis by cycloheximide (CHX) blocked the sodium chloride-induced alphaENaC mRNA downregulation, 3h after treatment. This indicates that synthesis of new, uncharacterized protein(s) is required for sodium chloride-mediated inhibition of alphaENaC gene transcription. PMID- 15120425 TI - Mechanisms of resistance to the cytotoxic effects of oxysterols in human leukemic cells. AB - We have developed hematopoietic cells resistant to the cytotoxic effects of oxysterols. Oxysterol-resistant HL60 cells were generated by continuous exposure to three different oxysterols-25-hydroxycholesterol (25-OHC), 7-beta hydroxycholesterol (7beta-OHC) and 7-keto-cholesterol (7kappa-C). We investigated the effects of 25-OHC, 7beta-OHC, 7kappa-C and the apoptotic agent staurosporine on these cells. The effect of the calcium channel blocker nifedipine on oxysterol cytotoxicity was also investigated. Differential display and real-time PCR were used to quantitate gene expression of oxysterol-sensitive and -resistant cells. Our results demonstrate that resistance to the cytotoxic effects of oxysterols is relatively specific to the type of oxysterol, and that the cytotoxicity of 25-OHC but not that of 7beta-OHC and 7kappa-C, appears to occur by a calcium dependent mechanism. Oxysterol-resistant cells demonstrated no significant difference in the expression of several genes previously implicated in oxysterol resistance, but expressed the bcl-2 gene at significantly lower levels than those observed in control cells. We identified three novel genes differentially expressed in resistant cells when compared to HL60 control cells. Taken together, the results of this study reveal potentially novel mechanisms of oxysterol cytotoxicity and resistance, and indicate that cytotoxicity of 25-OHC, 7beta-OHC and 7kappa-C occur by independent, yet overlapping mechanisms. PMID- 15120426 TI - The search to find the "magical" solution that will make our municipal solid wastes essentially disappear. PMID- 15120427 TI - Material and heavy metal balance in a recycling facility for home electrical appliances. AB - Collection and recycling of home electrical appliances was started in Japan in 2001 under a new recycling law. The law is aimed at promoting material recycling and at reducing the amount of waste to be landfilled. End of life products are processed by manual disassembly, shredding, and separation in 38 recycling facilities. The authors conducted a questionnaire survey and interviewed at some facilities to obtain information on process flow and material balance. By using the detailed records offered by one facility and by estimating the composition of recovered components, the material balance in the facilities was determined for four typical recycling processes. The heavy metal content of the recovered components was analyzed, then metal flow in the process was determined for each scenario. As a result, it was concluded that emissions to the environment of most heavy metals have been substantially reduced by the new recycling system, while a modest improvement in the rate of material recovery has been achieved. PMID- 15120428 TI - Resident's concerns and attitudes towards Solid Waste Management facilities. AB - Because of limited space, the siting and construction of a new SWM facility is a big challenge in Japan. An SWM facility should be socially accepted as well as environmentally and economically sound. This study aimed to investigate people's concerns about SWM facilities and their attitudes towards such facilities. A questionnaire was designed based on literature reviews and was sent to residents in three municipalities with different backgrounds. The questions covered concerns on the impact of an SWM facility, management aspects, unfairness of facility siting, and attitudes to facility construction. Of the many concerns, "pollution and health effect" had the highest rating, followed by "reliability", "damage to nature" and "cost". The rating was different between municipalities, reflecting their geographic and social backgrounds. Using factor analysis, correlations among concerns were analyzed, and five principal components were extracted, namely "pollution", "nuisance", "facility management", "planning of facility", and "merit/demerit". Although obvious correlations were not found between individual items of concern and attitudes to construction of a facility, the discriminant analysis indicated dominant concerns of attitudes, but the disagreement between actual impact and citizens were found. As for attributes, the "opposed" attitude decreased for residents who had visited an SWM facility, even if they had only seen it from outside. PMID- 15120429 TI - Stochastic modelling of landfill leachate and biogas production incorporating waste heterogeneity. Model formulation and uncertainty analysis. AB - A mathematical model simulating the hydrological and biochemical processes occurring in landfilled waste is presented and demonstrated. The model combines biochemical and hydrological models into an integrated representation of the landfill environment. Waste decomposition is modelled using traditional biochemical waste decomposition pathways combined with a simplified methodology for representing the rate of decomposition. Water flow through the waste is represented using a statistical velocity model capable of representing the effects of waste heterogeneity on leachate flow through the waste. Given the limitations in data capture from landfill sites, significant emphasis is placed on improving parameter identification and reducing parameter requirements. A sensitivity analysis is performed, highlighting the model's response to changes in input variables. A model test run is also presented, demonstrating the model capabilities. A parameter perturbation model sensitivity analysis was also performed. This has been able to show that although the model is sensitive to certain key parameters, its overall intuitive response provides a good basis for making reasonable predictions of the future state of the landfill system. Finally, due to the high uncertainty associated with landfill data, a tool for handling input data uncertainty is incorporated in the model's structure. It is concluded that the model can be used as a reasonable tool for modelling landfill processes and that further work should be undertaken to assess the model's performance. PMID- 15120430 TI - Pyrolysis of tyres. Influence of the final temperature of the process on emissions and the calorific value of the products recovered. AB - A study was made of the pyrolysis of tyre particles, with the aim of determining the possibilities of using the products resulting from the process as fuel. Three final temperatures were used, determined from thermogravimetric data. The design of the experiment was a horizontal oven containing a reactor into which particles of the original tyre were placed. After the process, a solid fraction (char) remained in the reactor, while the gases generated went through a set of scrubbers where most of the condensable fraction (oils) was retained. Finally, once free of this fraction, the gases were collected in glass ampoules. Solid and liquids fractions were subjected to thermogravimetric analyses in order to study their combustibility. The gas fraction was analysed by means of gas chromatography to establish the content of CO, CO2, H2 and hydrocarbons present in the samples (mainly components of gases produced in the pyrolysis process). A special study was made of the sulphur and chlorine content of all the fractions, as the presence of these elements could be problematic if the products are used as fuel. Tyre pyrolysis engenders a solid carbon residue that concentrates sulphur and chorine, with a relatively high calorific value, although not so high as that of the original tyre. The liquid fraction produced by the process has a high calorific value, which rises with the final temperature, up to 40 MJ/kg. The chlorine content of this fraction is negligible. Over 95% of the gas fraction, regardless of the final temperature, is composed of hydrocarbons of a low molecular weight and hydrogen, this fraction also appearing to be free of chlorine. PMID- 15120431 TI - Immobilization of lead and zinc in scrubber residues from MSW combustion using soluble phosphates. AB - The immobilization of MSWI-scrubber residues with soluble PO4(3-) was studied and compared to the immobilization using cement. The DIN 38414-S4 leaching protocol and pH dependent leaching were used to evaluate the leaching of Pb and Zn. Four different scrubber residues from MSW combustion (Pb concentration: 2.8-4.8 mg/g; Zn concentration: 3.0-12.3 mg/g) were mixed with water and cement or Na2HPO4 as source of soluble PO4(3-) at dosages of at least 0, 0.1, 0.2, 0.3 and 0.4 g per g residue. With cement as well as with PO4(3-) a reduction in Pb and Zn leaching was observed. With 0.4 g cement per g residue, the Pb leaching was reduced by a factor ranging from 70 to 100, but still exceeded the Pb landfill limit of 2 mg/l. With PO4(3-) the Pb leaching was reduced with a factor of 100-300 to below 2 mg/l. The Zn landfill limit (10 mg/l) was only exceeded by one untreated residue. Adding 0.2 g cement or 0.1 g PO4(3-) per g of that residue was enough to reduce leaching below 10 mg/l. However, when 0.6 g Na2HPO4 per g residue was added to a lime based scrubber residue, an increase in Zn leaching up to 12.5 mg/l was observed due to an increase in pH of up to 13.0. When using NaH2PO4 and H3PO4 no such increase in Zn leaching was observed. pH dependent leaching performed on one of the four residues showed that in the pH range of 2.5-6, Pb leaching was 100-50 times lower with Na2HPO4 treatment than with cement. In the pH range from 7-11, almost equal results were obtained for cement treated and Na2HPO4 treated residue. Above a pH of 12, Pb leaching was three times lower for the PO(4)(3-)-treated residue than for the cement treated residue. With soluble PO4(3-), Pb leaching below 2 mg/l could be attained at a dosage of 0.27 g PO4(3 )/g residue. With cement, Pb leaching was never below 2 mg/l. PMID- 15120432 TI - Removal of Cr(VI) from contaminated soil by electrokinetic remediation. AB - A new process for the removal of hexavalent chromium [Cr(VI)] contaminated soil is described. The process provides for an efficient removal of anionic chemicals from contaminated soils. Chromate anions were removed from the soil to the anodic reservoir by the moving force of electromigration. In this process, the chromate anions that accumulate in the anodic reservoir are simultaneously eliminated by using a column packed adsorbent. The adsorbent (immobilized tannin) used was chemically incorporated into cellulose. Cr(VI) was found to be adsorbed to this adsorbent efficiently. In the electrokinetic process, the pH of the aqueous solution in the anodic reservoir was decreased by the electrolysis of water. In the present study, the pH of the solution in the anodic reservoir is maintained at pH 6 by the addition of an aqueous alkaline solution during the electrokinetic process. The advantage of pH control is that it promotes the release of Cr(VI) from the soil by electromigration, thus permitting the maximum adsorption of Cr(VI) on the immobilized tannin. Simultaneous collection of Cr(VI) from the anodic reservoir leads to the protection from secondary contamination with Cr(VI). PMID- 15120433 TI - Modification of hydraulic conductivity in granular soils using waste materials. AB - This paper evaluates the use of waste products such as silica fume and fly ash in modification of the granular soils in order to remove some environmental problems and create new useful findings in the field of engineering. It is known that silica fume and fly ash, as well as clay material, are used in geotechnical engineering because of their pozzolanic reactivity and fineness to improve the soil properties needed with respect to engineering purposes. The main objective of this research project was to investigate the use of these materials in geotechnical engineering and to improve the hydraulic properties of soils by means of grouting. For this reason, firstly, suitable grouts in suspension forms were prepared by using silica fume, fly ash, clay and cement in different percentages. The properties of these cement-based grouts were then determined to obtain the desired optimum values for grouting. After that, these grouts were penetrated into the soil samples under pressure. The experimental work indicates that these waste materials and clay improved the physical properties and the fluidity of the cement-based grouts and they also decreased the hydraulic conductivity of the grouted soil samples by sealing the voids of the soil. The results of this study have important findings concerning the use of these materials in soil treatment and the improvement of hydraulic conductivity of the soils. PMID- 15120434 TI - Utilization of industrial by-products for the production of controlled low strength materials (CLSM). AB - Industrial by-products were used for the production of controlled low-strength material (CLSM). CLSM, also known as 'flowable fill' is used as a replacement of compacted soil in cases where the application of the latter is difficult or impossible. The low mechanical requirements (compared with structural concrete) enable the use of industrial by-products for the production of CLSM. In this study cement kiln dust, asphalt dust, coal fly ash, coal bottom ash and quarry waste were tested for the possibility of producing CLSM with large proportions of those wastes. The results showed that in most cases, CLSM with good properties could be made with significant amounts of dust (25-50%w), especially when the dust has some cementing or pozzolanic potential as do fly ash and cement kiln dust. PMID- 15120435 TI - Polyalkylphenyl-sulphonic acids with acid groups of variable strength from animal vegetable wastes. AB - The insoluble organic fraction isolated from rice-hulls residues and animal fecal matter mixture is sulphonated in liquid SO3 at 200 degrees C to a water soluble sulphonate (III). III is compared to the sulphonate (IV) obtained from the same mix after composting. Both products have been found to contain mixtures of molecules with close molecular weight. These molecules consist of a central cicloaliphatic cluster with peripheral pending aromatic chains. III and IV appear to have the same sulphonation degree. However, the latter contains higher concentrations of cicloaliphatic fragments and of amide, phenol and ether bonds, but less carboxylic and amine functional groups. These differences may be reasonably traced back to the starting materials. By comparison with commercial lignosulphonates derived from the paper and pulp industry, the above arylsulphonates are likely candidates for a variety of applications in the chemical industry and in agriculture. We conclude that sulphonation, even under the drastic experimental conditions of this work, does not seem to erase the memory of the parent matter structure. This reaction is capable of upgrading recalcitrant organic matter in vegetable waste residues to an interesting variety of lignosulphonates. PMID- 15120436 TI - Ensilage of pineapple processing waste for methane generation. AB - Pineapple peel wastes, which are seasonal, comprise of peels and rags. Their disposal poses a serious environmental pollution problem. Since pineapple peel is rich in cellulose, hemicellulose and other carbohydrates it was found to be a potential substrate for methane generation by anaerobic digestion. Ensilaging of pineapple peel resulted in the conversion of 55% carbohydrates into volatile fatty acids. The ensilage of pineapple processing wastes reduced the biological oxygen demand by 91%. Biogas digester fed with ensilaged pineapple peel resulted in the biogas yield of 0.67 m3/kg volatile solids (VS) added with methane content of 65% whereas fresh and dried pineapple peels gave biogas yields of 0.55% and 0.41 m3/kg VS added and methane content of 51% and 41% respectively. PMID- 15120437 TI - Towards an improved understanding of the leaching behavior of MSWI residues- Report on the 2nd meeting of the IWWG pHOENIX working group on "Management of Municipal Solid Waste Incineration Residues". PMID- 15120438 TI - Phyllanthus piscatorum, ethnopharmacological studies on a women's medicinal plant of the Yanomami Amerindians. AB - The shrub Phyllanthus piscatorum Kunth (Euphorbiaceae) is cultivated by various ethnic groups of the Amazon because of its piscicidal properties. During ethnobotanical fieldwork among the Yanomami Amerindians in Venezuela we observed that Phyllanthus piscatorum was exclusively cultivated and used by the women. Aerial parts of this herbaceous shrub are employed as fish poison and medicine to treat wounds and fungal infections. In addition, the leaves are used as tobacco substitute. Ethnobotanical data regarding the context of the use of this plant are presented. To validate ethnobotanical information related to its medicinal indications, antimicrobial, and antiprotozoal properties of water, methanol (MeOH) and dichloromethane (DCM) extracts were studied. No activity against Gram positive bacterial strains but significant activity against the fungi Aspergillus fumigatus, Aspergillus flavus and the yeast Candida albicans were found. All extracts showed weak in vitro activity against Plasmodium falciparum and Trypanosoma brucei rhodesiense. The extracts were further investigated for cytotoxic effects in an in vitro test system with leukemia Jurkat T, HeLa, and human peripheral mononuclear blood cells (PBMCs). During the first 48 h the extracts did not exhibit any cytotoxicity. After 72 h the DCM extract potently inhibited viability of HeLa cells. Although in several communities along the upper Orinoco the cultivation and use of Phyllanthus piscatorum is being lost because of the ongoing acculturation, the traditional medicinal use of Phyllanthus piscatorum might provide an effective and cheap remedy against dermatological diseases linked with Candida albicans infections. PMID- 15120439 TI - Pharmacopoeia in a shamanistic society: the Izoceno-Guarani (Bolivian Chaco). AB - We present the results of an ethnopharmacological research within a Bolivian lowland ethnic group from the dry Chaco, the izoceno-guarani. Izoceno-guarani people belong to the extended Chiriguano group. They are actually organised in independent communities, settled down in south-east Bolivia. Struggling very soon for their rights, landowners of their territory, izoceno-guarani appear to be well organised and maintain a still vivid culture. Medicine is in the hands of Paye who are recognised as specialists in their own group. Ethnopharmacological research leads us to collect approximately over 306 species, 189 of them having medicinal uses. We present here an overview of the izoceno-guarani ethnomedicine and pharmacopoeia, based on vegetal and animal products. PMID- 15120440 TI - Antidiabetic and antihyperlipidaemic effect of alcoholic Syzigium cumini seeds in alloxan induced diabetic albino rats. AB - Syzigium cumini, commonly known as 'jamun', is widely used in different parts of India for the treatment of diabetes mellitus. The present study was designed to evaluate the antidiabetic and antihyperlipidaemic effect of an alcoholic extract of Syzigium cumini seeds (JSEt) in alloxan diabetic rats. Diabetes was induced by single intraperitoneal injection of alloxan (150 mg kg(-1) body weight). Oral administration of alcoholic JSEt to diabetic rats at a dose of 100 mg kg(-1) body weight resulted in a significant reduction in blood glucose and urine sugar and lipids in serum and tissues in alloxan diabetic rats. The extract also increases total haemoglobin. The extract brought back all the parameters to normal levels. The effect of alcoholic JSEt was similar to that of insulin. Thus, our investigation clearly shows that alcoholic JSEt has both antidiabetic and antihyperlipidaemic effects. PMID- 15120441 TI - Anti-inflammatory and analgesic activity of Peperomia pellucida (L.) HBK (Piperaceae). AB - An aqueous extract of the aerial part of Peperomia pellucida (L.) HBK (Piperaceae) was tested for anti-inflammatory (paw edema induced by carrageenin and arachidonic acid) and analgesic activity (abdominal writhes and hot plate) in rats and mice, respectively. Oral administration of 200 and 400 mg/kg of the aqueous extract exhibited an anti-inflammatory activity in the carrageenin test, which was based on interference with prostaglandin synthesis, as confirmed by the arachidonic acid test. In the abdominal writhing test induced by acetic acid, 400 mg/kg of the plant extract had the highest analgesic activity, whereas in the hot plate test the best dose was 100 mg/kg. The LD(50) showed that Peperomia pellucida (5000 mg/kg) presented low toxicity. PMID- 15120442 TI - Diuretic and nephroprotective effect of Jawarish Zarooni Sada--a polyherbal unani formulation. AB - A number of drugs, both single and compound preparations are used widely in Tibb e-Unani (Unani medicine) in the management of renal diseases. But such drugs mostly, have not been investigated for their described effects. Jawarish Zarooni Sada (JZS) is one such polyherbal preparation containing 15 ingredients, mainly described to be diuretic and nephroprotective. Therefore, in the present study ethanol and water extracts of JZS (300 mg each) were investigated for diuretic activity by measuring the total urine output over a period of 6h. Sodium and potassium level in urine sample was also estimated. Nephroprotective activity of JZS against gentamicin-induced nephrotoxicity was investigated by administering JZS along with high dose of gentamicin (40 mg/kg) and elevation of serum urea and serum creatinine was taken as the index of nephrotoxicity. JZS showed significant diuretic and nephroprotective effect. PMID- 15120443 TI - Inductive effect of the leaf mixture extract of Aloe buettneri, Justicia insularis, Dicliptera verticillata and Hibiscus macranthus on in vitro production of estradiol. AB - In the course of a preliminary clarification of the mechanisms of the leaf mixture extract of Aloe buettneri, Justicia insularis, Dicliptera verticillata and Hibiscus macranthus, locally used to regulate the menstrual cycle and to treat dysmenorrhea or cases of infertility in women, pieces of proestrus rat ovary were incubated in the presence of increasing concentration of the plant extract and/or human Chorionic Gonadotropin (hCG). The in vitro production of estradiol and progesterone by ovarian cells of proestrus rat was significantly increased in the presence of various concentration of hCG (P < 0.05). The different concentration of the plant extract increased the production of estradiol by twofold. In addition, the in vitro production of estradiol by ovarian cells increased by 13-fold when they were incubated with hCG (0.1 IU/ml) and a concentration of 130 microg/ml of the plant extract. These results clearly attest the direct effects of some chemical components of the leaf mixture of the plants on ovarian steroidogenesis. PMID- 15120444 TI - Cholesterol lowering effect of SG-GN3, the extract of salted and fermented small shrimps, Acetes japonicus, in Triton WR-1339 or high cholesterol-diet induced hypercholesterolemic rats. AB - The cholesterol lowering effect of SG-GN3, the extract of salted and fermented small shrimps, Acetes japonicus, was investigated in hypercholesterolemic animal models. Hypercholesterolemia was induced with Triton WR-1339 (nonionic detergent) or high cholesterol (HC)-diet. SG-GN3 significantly decreased total cholesterol (TC) in Triton WR-1339 model at 30 post-treatment hour (549.80 +/- 152.46 mg/dl) compared to the control which induced by only Triton WR-1339 (798.84 +/- 94.98 mg/dl), whereas high-density lipoprotein (HDL) content did not decrease (P < 0.05). In HC-diet model, TC content significantly decreased by SG-GN3 treatment at 3 post-treatment day (P < 0.05). These results suggest that SG-GN3 effectively decreased serum TC level in hypercholesterolemic animal models. PMID- 15120445 TI - Anti-inflammatory activity of methanolic extracts from Ventilago harmandiana Pierre. AB - Methanolic extracts from the heart wood, stem bark, and stem wood of Ventilago harmandiana Pierre (Family Rhamnaceae) were assessed for anti-inflammatory effects using both acute and chronic inflammatory models. Analgesic and antipyretic activities of the extracts were also evaluated. It was found that all extracts possessed strong inhibitory effects on the acute phase of inflammation as seen in ethyl phenylpropiolate (EPP)- and arachidonic acid (AA)-induced ear edema as well as in carrageenin-induced paw edema in rats. The extracts elicited only weak inhibitory activity on cotton pellet-induced granuloma formation, a subchronic inflammatory model. In the analgesic test, all extracts exerted pronounced inhibitory activity in acetic acid-induced writhing response but showed only weak effects in the tail-flick test. The extracts also showed excellent antipyretic activity on yeast-induced hyperthermia in rats. PMID- 15120446 TI - Antiulcer activity of Utleria salicifolia rhizome extract. AB - The effect of 50% ethanolic extract of Utleria salicifolia (USE) was assessed in different acute and chronic gastric ulcer models in rats. USE, 50-200 mg/kg administered orally, twice daily for 5 days showed dose-dependent ulcer protective effect in pylorus ligation (14.48-51.03% protection, P < 0.5 to P < 0.01), aspirin (28.80-56.52% protection, P < 0.5 to P < 0.001), ethanol (13.22 60.74% protection, P < 0.5 to P < 0.001), cold-restraint stress (21.22-77.14% protection, P < 0.05 to P < 0.001), and acetic acid (20.0-84.37% protection, P < 0.5 to P < 0.001)-induced acute and chronic ulcers. USE also significantly (P < 0.001) reduced the ulcer incidence (50 and 10%) and severity (67.83 and 91.34% protection) of duodenal ulcer, induced by cysteamine. Besides USE offered protection (53.52 and 60.58%) against ethanol-induced depletion of gastric wall mucus. However, USE reduced the ulcer index with significant decrease in plasma corticosterone (25.53 and 39.52% protection, P < 0.1 and P < 0.05), lipid peroxidation (18.75 and 47.92% protection, P < 0.01 and P < 0.001), superoxide dismutase (15.80 and 26.61% protection, P < 0.05 and P < 0.001) and increased in catalase (28.42 and 71.0% protection, P < 0.05 and P < 0.001) activity, respectively. Preliminary phytochemical screening of the USE gave the positive test for steroids, alkaloids, terpenoids, saponins and tannins. The HPTLC studies in the toluene: ethyl acetate: formic acid and the densitometric scanning at 254 nm gave three major spots with area corresponding to 28.16, 17.17, and 13.79% at 0.69, 0.78, and 0.88 R(f) values, respectively. The results indicate that USE possesses antiulcer activity. PMID- 15120447 TI - Immunoadjuvant potential of Asparagus racemosus aqueous extract in experimental system. AB - The immunoadjuvant potential of Asparagus racemosus (Willd.) Family (Liliaceae) aqueous root extract was evaluated in experimental animals immunized with diphtheria, tetanus, pertussis (DTP) vaccine. Immunostimulation was evaluated using serological and hematological parameters. Oral administration of test material at 100 mg/kg per day dose for 15 days resulted significant increase (P = 0.0052) in antibody titers to Bordtella pertussis as compared to untreated (control) animals. Immunized animals (treated and untreated) were challenged with B. pertussis 18323 strain and the animals were observed for 14 days. Results indicate that the treated animals did show significant increase in antibody titers as compared to untreated animals after challenge (P = 0.002). Immunoprotection against intra-cerebral challenge of live B. pertussis cells was evaluated based on degree of sickness, paralysis and subsequent death. Reduced mortality accompanied with overall improved health status was observed in treated animals after intra-cerebral challenge of B. pertussis indicating development of protective immune response. Present study indicates applications of test material as potential immunoadjuvant that also offers direct therapeutic benefits resulting in less morbidity and mortality. PMID- 15120448 TI - The effects of bitter melon (Momordica charantia) on serum and liver triglyceride levels in rats. AB - Effects of three different varieties (Koimidori, Powerful-Reishi, and Hyakunari) of bitter melon (Momordica charantia) and those of methanol fraction extract of Koimidori variety on serum and liver triglycerides were studied in rats. Feeding of diets containing either bitter melon or various fractions isolated by organic solvents caused no adverse effects on food intake or growth of rats. When the effect of three different varieties of bitter melon was compared, the Koimidori variety was found to be the most effective in lowering hepatic triglyceride levels as compared to the other two varieties, suggesting a variety-dependent difference in their activity. Furthermore, the active component(s) responsible for the liver triglyceride lowering activity of Koimidori variety was assumed to be concentrated in the methanol fraction, but not in other fractions such as the n-hexane, the acetone, or the residual fraction. The triglyceride lowering activity was furthermore confirmed by the dose-dependent reduction of hepatic triglyceride, resulting the lowest level in rats fed 3.0% supplementation. In these experiments, the effects on serum lipids were marginal. The results of the present and previous studies clearly show that bitter melon, especially Koimidori variety, exhibits a potent liver triglyceride-lowering activity. PMID- 15120449 TI - Vasodilator activity of Michelia figo Spreng. (Magnoliaceae) by in vitro functional study. AB - The methanolic extract of leaves of Michelia figo Spreng. (Magnoliaceae), as well as several purified fractions, showed a concentration-dependent vasorelaxing effect on aortic rings endothelium-deprived and pre-contracted by norepinephrine (NE). For further pharmacological investigation on the mechanism of action, the fraction S4 was selected, since it showed the best vasodilator properties. The pharmacological effect was not produced through the stimulation of cyclooxygenase, adenyl cyclase, or guanylyl cyclase, since selective inhibitors did not prevent the fraction S4-induced effects. Moreover, the vasorelaxing effect of the fraction was resistant to the block of nifedipine-sensitive Ca(2+) channels. The fraction S4 (10(-4) g/ml) produced a shift towards the right of the concentration-contractile response curve to NE, in normal conditions, and the shift was more evident in Ca(2+)-free Tyrode solution, suggesting an action on intracellular Ca(2+)-channels. The vasodilator action of fraction S4 on NE pre contracted rings was not prevented by cyclopiazonic acid (blocker of Ca(2+)/ATPase), which excludes a role for mechanisms involving the storage of Ca(2+) in the sarcoplasmic reticulum. The reduction of the contraction elicited by caffeine, an opener of ryanodine-sensitive receptors, suggests that the fraction S4 of Michelia figo leaves could produce the vasorelaxing response by the blockade of ryanodine-sensitive Ca(2+) channels of the sarcoplasmic reticulum. PMID- 15120450 TI - Salix caprea inhibits skin carcinogenesis in murine skin: inhibition of oxidative stress, ornithine decarboxylase activity and DNA synthesis. AB - Chemoprevention of free radical-mediated diseases including cancer by natural products is an emerging discipline due to its wider applicability and acceptance. The present study deals with the chemopreventive effect of Salix caprea against phorbol ester-induced oxidative stress and tumor promotion in murine skin. In the present investigation, it was observed that a single application of 12-O tetradecanoyl-13-phorbol acetate (TPA) (20 nmol/0.2 ml acetone/animal) caused a significant (P < 0.05) depletion of cutaneous antioxidants viz., glutathione, glutathione reductase, glutathione peroxidase, catalase and phase II drug metabolizing enzymes viz., glutathione-S-transferase, quinone reductase. An increase in the hydrogen peroxide generation and protein oxidation (measured in terms of protein carbonyl content) was also observed with a single application of TPA. However, the pretreatment of animals with different doses of Salix caprea (0.5, 1.0 and 1.5 mg/kg/0.2 ml acetone) caused a significant recovery in the TPA mediated depletion in antioxidant levels. The pretreatment of animals with Salix caprea was observed to inhibit the TPA-mediated depletion in phase II enzymes. It was also observed that Salix caprea reversed the TPA-mediated depletion in the activity of phase II enzymes that is an important characteristic of cancer chemopreventive agents. Phorbol esters are known to induce the tumor promotion by increasing rate of DNA synthesis, ornithine decarboxylase activity (ODC), and xanthine oxidase activity. In the present investigation, it was observed that the pretreatment of animals with Salix caprea caused a significant (P < 0.05) depletion in the TPA-induced DNA synthesis, ODC and xanthine oxidase activity in mice skin. Salix caprea significantly reduced the tumor promotion in mice skin when tested in two-stage chemical carcinogenesis model. It was observed to inhibit significantly P < 0.05) the 7,12-dimethyl benz[a] anthracene (DMBA) initiated phorbol ester promoted skin carcinogenesis. It was concluded from the results that Salix caprea is an effective antioxidant and chemopreventive agent against phorbol ester-induced tumor promotion. PMID- 15120452 TI - The herbal market of Thessaloniki (N Greece) and its relation to the ethnobotanical tradition. AB - The results of a survey of the medicinal plants found in the herbal market of Thessaloniki, which comprises traditional shops, modern shops and open-air market stalls, are presented. A total number of 172 taxa, Pteridophyta and Spermatophyta, were found in 18 selected market spots. Information is provided on the origin, the plant parts used, the ways of drug preparation and the medicinal uses of the herbs found. The majority of them (133 taxa) are of Greek origin and are gathered from the wild (99). A remarkable number of herbs (93) found in the market of Thessaloniki are mentioned by Dioscurides whereas the comparison to the recent ethnobotanical information shows that the utilization of Dioscurides' plants remains uninterrupted. Thus it is suggested that the herb trade is still based on the Greek ethnobotanical tradition, dating from antiquity. PMID- 15120451 TI - Pharmacokinetic and pharmacodynamic studies on interaction of "Trikatu" with diclofenac sodium. AB - "Trikatu"-an Ayurvedic formulation comprising of a 1:1:1 ratio of dried fruits of Piper nigrum, Piper longum and dried rhizomes of Zingiber officinale is widely used to enhance the bioavailability of drugs, like vasicine, indomethacin, etc. The enhanced biological response might lead to alteration of therapeutic regimens of commonly prescribed drugs. The present work was aimed to study the effect of concomitant administration of Trikatu on the pharmacokinetics and pharmacodynamics of diclofenac sodium, a frequently prescribed non-steroidal anti inflammatory drug, having a poor oral bioavailability (54 +/- 2%). The effect of Trikatu on the bioavailability profile of diclofenac sodium was studied in rabbits. It was observed that Trikatu significantly decreased the serum levels of diclofenac sodium. The pharmacodynamic study was carried out to evaluate the effect of Trikatu on the anti-inflammatory activity of diclofenac sodium using carragenin-induced rat paw edema model. It was observed that the mean percent edema inhibition shown by the combination of Trikatu and diclofenac was similar to that shown by Trikatu alone but significantly less than that shown by diclofenac alone. Thus, the experimental findings indicated that Trikatu pretreatment might decrease the bioavailability of certain drugs probably through a drug-herb interaction thereby adversely affecting the therapeutic efficacy of these drugs. PMID- 15120453 TI - Analgesic and antiinflammatory activities of Erigeron floribundus. AB - This study was intended to evaluate the analgesic and antiinflammatory activities of an aqueous extract of Erigeron floribundus (H.B. & K) or (syn": Conyza sumatrensis (Retz) E.K. Walker) (Asteraceae). Phytochemical analysis was carried out using standard methodologies. The analgesic investigations were carried out against two types of noxious stimuli, chemical (formalin-induced pain and acetic acid-induced writhing) and thermal (hotplate and tail immersion tests). The effects following aspirin and naloxone pretreatments were also studied. For the antiinflammatory activities, the carrageenan-induced oedema of the hindpaw of rats was used and the paw volume measured plethysmometrically from 0 to 24 h after injection. This was compared to a standard drug indomethacin (10 mg/kg). The results were subjected to statistical analysis. The plant had saponins, flavonoids, glycosides, alkaloids, oils, phenols and tannins and significantly increased the reaction time of hotplate and immersion tests. It decreased the writhings of acetic acid-induced abdominal contractions and lickings of formalin induced pain. Aspirin had no effect on hotplate and tail immersion tests but showed an effect on writhing test. These results showed that the plant had both central and peripheral acting effects and this was confirmed by its effect on both phases of formalin-induced pain. The extract also significantly decreased the rat paw oedema volume at 50 mg/kg and above. In conclusion, Erigeron floribundus has central and peripheral analgesic properties as well as antiinflammatory activities. PMID- 15120454 TI - Study of the hypoglycaemic activity of Fraxinus excelsior and Silybum marianum in an animal model of type 1 diabetes mellitus. AB - The hypoglycaemic effect of the aqueous extracts of Fraxinus excelsior (FE) seed and Silybum marianum (SM) aerial part was investigated in normal and streptozotocin (STZ) diabetic rats. After a single dose or 15 daily doses, oral administration of the aqueous extracts (20 mg/kg) produced a significant decrease of blood glucose levels in both normal and STZ diabetic rats (P < 0.001). From the first week, the body weight was increased in normal rats (P < 0.05) and decreased in STZ rats (P < 0.01) after FE administration. In addition, no changes were observed in basal plasma insulin concentrations after both FE and SM treatments in either normal and STZ diabetic rats indicating that these plants exert their pharmacological activity without affecting insulin secretion. We conclude that the aqueous extracts of FE and SM exhibit potent hypoglycaemic and anti-hyperglycaemic activities in normal and STZ rats, respectively, without affecting basal plasma insulin concentrations. PMID- 15120455 TI - Diuretic activity of Spilanthes acmella flowers in rats. AB - In the Sri Lankan traditional medicine, Spilanthes acmella Murr. (Family: Compositae) flowers are claimed to possess powerful diuretic activity. However, as yet, the diuretic potential of these flowers is not investigated by scientifically controlled studies. The aim of this study was to evaluate the diuretic potential of Spilanthes acmellaflowers in rats using a cold-water extract (CWE). Different concentrations of CWE (500, 1000, 1500 mg/kg) or vehicle or furosemide (13 mg/kg) were orally administered (N = 6 per each treatment group) to hydrated rats and their urine output was monitored at several intervals of time (1-5 h). The highest dose of CWE significantly (P < 0.05) and markedly increased the urine output. The onset of this diuretic action was extremely prompt (within 1 h) and lasted throughout the studied period (up to 5 h). The peak effect was evident between 1 and 2 h. Further, the intensity of diuresis induced by the CWE in the first hour was almost similar to that of furosemide. Spilanthes acmella CWE also caused marked increase in urinary Na(+) and K(+) levels and a reduction in the osmolarity of urine suggesting that it is mainly acting as a loop diuretic. It may also inhibit ADH release and/or action. It is concluded that the Spilanthes acmella CWE has strong diuretic action as is claimed. PMID- 15120456 TI - Antispasmodic and hypotensive effects of Ferula asafoetida gum extract. AB - The effects of Ferula asafoetida gum extract on the contractile responses of the isolated guinea-pig ileum induced by acetylcholine, histamine and KCl, and on the mean arterial blood pressure of rat were investigated. In the presence of extract (3 mg/ml), the average amplitude of spontaneous contractions of the isolated guinea-pig ileum was decreased to 54 +/- 7% of control. Exposure of the precontracted ileum by acetylcholine (10 microM) to Ferula asafoetida gum extract caused relaxation in a concentration-dependent manner. Similar relaxatory effect of the extract was observed on the precontracted ileum by histamine (10 microM) and KCl (28 mM). However, when the preparations were preincubated with indomethacin (100 nM) and different antagonists, such as propranolol (1 microM), atropine (100 nM), chlorpheniramine (25 nM) then were contracted with KCl, exposure to the extract (3 mg/ml) did not cause any relaxation. Furthermore, Ferula asafoetida gum extract (0.3-2.2 mg/100g body weight) significantly reduced the mean arterial blood pressure in anaesthetised rats. It might be concluded that the relaxant compounds in Ferula asafoetida gum extract interfere with a variety of muscarinic, adrenergic and histaminic receptor activities or with the mobilisation of calcium ions required for smooth muscle contraction non specificly. PMID- 15120457 TI - Evaluation of acute and chronic treatments with Harpagophytum procumbens on Freund's adjuvant-induced arthritis in rats. AB - The extract of Harpagophytum procumbens, widely utilized in Europe and, more recently, in other countries, is traditionally indicated to treat inflammatory processes. Harpagophytum procumbens acts by way of interleukins and leukocyte migration to the painful and inflamed joint area. Chemically, its secondary tuberous roots contains iridoid glycosides, harpagogide, procumbide, and harpagoside, as the active principle. The purpose of the present study was evaluate the therapeutic potential as anti-inflammatory and analgesic agent in rat model of Freund's adjuvant-induced arthritis both in the acute and chronic phases. The animals were injected with Freund's adjuvant in sub-plantar tissue of the right posterior paw and randomly assigned in acute (25, 50, or 100 mg/kg) or chronic (100 mg/kg) treatments with Harpagophytum procumbens solution test or vehicle. Then, submitted to behavioral test and assessment of body weight and right paw's measurements. The results show that Harpagophytum procumbens extract increased the animals 'latency of paws' withdrawal, indicating a protective effect against the pain induced by the thermal stimulus, both in acute and chronic treatments. In addition to reduction in the right paw edema in the experimental groups when compared to control group. Thus, the data showed anti inflammatory and peripheral analgesic properties of Harpagophytum procumbens extract with all doses tested, thus confirming its indication for inflammatory processes. PMID- 15120458 TI - Ethnopharmacognostic survey on the natural ingredients used in folk cosmetics, cosmeceuticals and remedies for healing skin diseases in the inland Marches, Central-Eastern Italy. AB - An ethnopharmaceutical study focused on domestic cosmetics, cosmeceuticals, and remedies to heal skin diseases traditionally used in the inland part of the Marches region (Central-Eastern Italy) has been conducted. At present, traditional knowledge concerning home-made phytocosmetics is represented by both the remnants of an orally transmitted folk heritage and also by new forms of knowledge, sometimes coming from popular phytotherapeutical books and the mass media (out of the scope of this survey), but also as a result of recent migration trends from Eastern Europe. We recorded approximately 135 cosmetic or cosmeceutical preparations prepared from more than 70 botanical species and a very few animal or mineral ingredients. Among the recorded preparations, developing a clear distinction amongst cosmetics, cosmeceuticals and pharmaceuticals for skin diseases is very problematic, confirming that in folk knowledge systems medicinal products for healing skin diseases and cosmetics have often been perceived as two poles of a continuum. Many of the quoted species represented well-known medicinal plants of the European phytotherapy, although we also recorded a few unusual plant taxa, which are briefly discussed under the perspective of their eventual phytochemical and/or phytopharmacological potentialities. Exotic drugs or precious essences, even native of the Mediterranean, were not quoted as ingredients for preparing perfumes and fragrances by the interviewees of the present study, thus indicating that popular cosmetic practices in rural Central Italy have taken a much separated path away from the cosmetic "know-how" of the aristocracy and high bourgeois classes of the last centuries. PMID- 15120459 TI - Antidepressant effect of three traditional Chinese medicines in the learned helplessness model. AB - Plantago asiatica, Scrophularia ningpoensis and Ilex pubescens are among the traditional Chinese medicines which are more frequently prescribed for treating depression-like ailments in the past and present traditional Chinese medical practice. The present work was therefore conducted to evaluate the presumable antidepressant effects of the extracts derived from the three remedies in mice using the learned helplessness model being used for screening for antidepressant compounds in modern medicinal researches. As a result, the petroleum extracts of Plantago asiatica and Ilex pubescens as well as the EtOAc extract of Scrophularia ningpoensis and the petroleum-soluble fraction of the acidic hydrolysate of the water extract of Ilex pubescens (after petroleum extraction) decreased significantly the number of escape failures relative to the control. The finding rationalized the clinical prescription of the herbs for the treatment of depression, and shined a clue for the characterization of the antidepressant phytochemical(s). PMID- 15120460 TI - Inhibition of MAO A and B by some plant-derived alkaloids, phenols and anthraquinones. AB - A total of seventeen phytochemicals including seven alkaloids (piperine, strychnine, brucine, stachydrine, tetrandrine, frangchinoline and sinomenine), four phenols (paeonol, honokiol, magnolol and eugenol) and six anthraquinones (emodin, rhein, chrysorphanol, aloe-emodin, physcion and 1,8 dihydroxyanthraquinone) was examined for inhibitory activity of monoamine oxidase (MAO) A and B from rat brain mitochondrial. Among these compounds, piperine and paeonol were found to be inhibitory against MAO A in a dose-dependent manner with IC(50) values of 49.3 and 54.6 microM, respectively. Piperine, paeonol and emodin were shown to inhibit MAO B in a dose-dependent manner with the IC(50) data of 91.3, 42.5 and 35.4 microM, respectively. Lineweaver-Burk transformation of the inhibition data indicated that the inhibitory action of piperine on MAO A was of mixed type, and that of paeonol on the same type of the enzyme was of non competitive type. For piperine, the K(i) and K(I) were determined to be 35.8 and 25.7microM, respectively. For paeonol, the K(i) was estimated to be 51.1 microM. The inhibition of piperine and paeonol on MAO B was of competitive type with K(i) values of 79.9 and 38.2 microM, respectively. The inhibition of emodin on MAO B was of mixed type with the K(i) and K(I) data of 15.1 and 22.9 microM, respectively. The present investigation showed that the phytochemicals piperine, paeonol and emodin are potent MAO inhibitors whereas other compounds were inactive against any type of MAO at 100 microM in the present assay. PMID- 15120461 TI - Effects of Piper longum fruit, Piper sarmentosum root and Quercus infectoria nut gall on caecal amoebiasis in mice. AB - The anti-amoebic effects of crude methanol extracts of Piper longum fruit, Piper sarmentosum root and Quercus infectoria nut gall against Entamoeba histolytica infecting the caecum of mice were studied. Caecal amoebiasis in mice was induced by injection of Entamoeba histolytica trophozoites directly into the caecum. The mice were then treated orally with the extract, a standard drug (metronidazole), or vehicle p.o. for five consecutive days, beginning 24 h after the infection and were examined on the sixth day. At a dose of 1000 mg/kg per day, the extracts of Piper longum fruit, Piper sarmentosum root and Quercus infectoria nut gall had a curative rate of 100, 40 and 26%, respectively. At a concentration of 500 and 250 mg/kg/day, extract from Piper longum fruit was still effective in 93 and 46% of the cases, respectively, while extract from Piper sarmentosum root at a dose of less than 1000 mg/kg per day did not cure any mice from amoebiasis. Extract of Quercus infectoria nut gall at a concentration of 500 and of 250 mg/kg per day cured 26 and 13% of mice, respectively. Metronidazole at a concentration of 125 and of 62.5 mg/kg per day had a curative rate of 100 and 60%, respectively. The severity of caecal wall ulceration was reduced in mice which received the extract and metronidazole as compared to the control animals. PMID- 15120462 TI - Memory enhancing activity of Glycyrrhiza glabra in mice. AB - In the traditional system of medicine, the roots and rhizomes of Glycyrrhiza glabra (family: Leguminosae) have been employed clinically for centuries for their anti-inflammatory, antiulcer, expectorant, antimicrobial and anxiolytic activities. The present study was undertaken to investigate the effects of Glycyrrhiza glabra (popularly known as liquorice) on learning and memory in mice. Elevated plus-maze and passive avoidance paradigm were employed to test learning and memory. Three doses (75, 150 and 300 mg/kg p.o.) of aqueous extract of Glycyrrhiza glabra were administered for 7 successive days in separate groups of animals. The dose of 150 mg/kg of the aqueous extract of liquorice significantly improved learning and memory of mice. Furthermore, this dose significantly reversed the amnesia induced by diazepam (1 mg/kg i.p.) and scopolamine (0.4 mg/kg i.p.). Anti-inflammatory and antioxidant properties of liquorice may be contributing favorably to the memory enhancement effect. Since scopolamine induced amnesia was reversed by liquorice, it is possible that the beneficial effect on learning and memory was due to facilitation of cholinergic-transmission in mouse brain. However, further studies are necessitated to identify the exact mechanism of action. In the present investigation, Glycyrrhiza glabra has shown promise as a memory enhancing agent in all the laboratory models employed. PMID- 15120463 TI - Anti-inflammatory evaluation of Ionidium suffruticosam Ging. in rats. AB - The anti-inflammatory activity of Ionidium suffruticosam (Violaceae) methanol extract was evaluated on carrageenin, histamine and serotonin-induced rat hind paw oedema acute models. The extract at doses of 200 and 400 mg/kg has been found to possess significant anti-inflammatory activity on the tested experimental models. The extract at the dose level of 400 mg/kg exhibited maximum anti inflammatory activity in all the animal models. In a chronic test, the extract (400 mg/kg) showed 42.78% reduction in granuloma weight. The effect produced by the extract was comparable to that of phenylbutazone, a proto type of a non steroidal anti-inflammatory agent. PMID- 15120464 TI - Hospitals, managed care, and the charity caseload in California. AB - We ask whether increasing HMO penetration causes hospitals to cut back on charity care using California hospital discharge data for 1988-1996. There is little evidence at the hospital level that private hospitals respond to HMOs by turning away uninsured and/or Medicaid patients. In the for-profit sector hospitals actually reduce the share of privately insured patients and increase the shares of Medicare patients and Medicaid births. Apparently, HMO penetration reduces the price paid by privately insured patients, making them relatively less attractive to for-profit hospitals. PMID- 15120465 TI - Estimating medical care costs under conditions of censoring. AB - A number of non-parametric estimators have been proposed to calculate average medical care costs in the presence of censoring. This paper assesses their performance both in terms of bias and efficiency under extreme conditions using a medical dataset which exhibits heavy censoring. The estimators are further investigated using artificially generated data. Their variances are derived from analytic formulae based on the estimators' asymptotic properties and these are compared to empirically derived bootstrap estimates. The analysis revealed various performance patterns ranging from generally stable estimators under all conditions considered to estimators which become increasingly unstable with increasing levels of censoring. The bootstrap estimates of variance were consistent with the analytically derived asymptotic variance estimates. Of the two estimators that performed best, one imposes restrictions on the censoring distribution while the other is not restricted by the censoring pattern and on this basis the second may be preferred. PMID- 15120466 TI - Quality and employers' choice of health plans. AB - We seek to understand the relationship between employer decisions regarding which health plans firms choose to offer to their employees and the performance of those plans. We measure performance using data from the Health Plan Employer Data Information Set (HEDIS) and the Consumer Assessment of Health Plan Survey (CAHPS). We use a unique data set that lists the Health Maintenance Organizations (HMOs) available to, and offered by, large employers across markets in the year 2000, and examine the relationship between plan offerings, performance measures and other plan characteristics. We estimate two sets of specifications that differ in whether they model plan choice as a function of absolute plan performance or plan performance relative to competitors. We find that employers are more likely to offer plans with strong absolute and relative HEDIS and CAHPS performance measures. Our results are consistent with the view that large employers are responsive to the interests of their employees. PMID- 15120467 TI - Teenage sex, drugs and alcohol use: problems identifying the cause of risky behaviors. AB - The relationship between substance use and adolescent sexual activity is an important one, and extensive literature has shown that substance use is positively associated with adolescent sexual behaviors. While this is true, causality from substance use to risky sexual behaviors is difficult to establish, as it is likely that an adolescent's sexual behavior and substance use depend on a set of personal and social behaviors, many of which are unmeasured. Researchers must thus devise a credible empirical strategy in order to overcome this omitted variable bias. Using the first waves of the National Longitudinal Survey of Adolescent Health and the 1997 National Longitudinal Survey of Youth, we call into question recent methods used to determine causality. Despite attempts to determine the causal relationship between substance use and sexual behavior, the nature of the relationship remains unknown. PMID- 15120468 TI - Measuring health inequality using qualitative data. AB - Many questions in health policy require an understanding of the distribution of health status across a given population and how it changes as a result of policy interventions. Since objective data on individual health status are often unavailable or incomplete, especially for populations with very low mortality, increasing use has been made of self-reported health status (SRHS) data, which record people's own perceptions of their health status. SRHS has been shown to be a strong predictor of objective health outcomes and indications, including mortality. Nevertheless, the qualitative or categorical nature of SRHS data prevents the straightforward use of traditional tools of distributional analysis, such as the Lorenz curve, in evaluating inequality. This paper presents a methodology for evaluating inequality when the data are qualitative rather than quantitative in nature. A partial inequality ordering is defined to indicate when a distribution is more "spread out" than another; a second partial ordering (first order dominance) is used to indicate when the overall health level rises. Both are applicable to qualitative data, such as SRHS, in that results do not depend on the numerical scaling assigned to the categories. The approach is illustrated using SRHS data from the National Health Interview Survey (NHIS) State Data Files for 1994, focusing on the distribution of SRHS within states. PMID- 15120469 TI - Too much ado about two-part models and transformation? Comparing methods of modeling Medicare expenditures. AB - Many methods for modeling skewed health care cost and use data have been suggested in the literature. This paper compares the performance of eight alternative estimators, including OLS and GLM estimators and one- and two-part models, in predicting Medicare costs. It finds that four of the alternatives produce very similar results in practice. It then suggests an efficient method for researchers to use when selecting estimators of health care costs. PMID- 15120470 TI - A social choice approach to expert consensus panels. AB - This study uses recent theoretical work about group decision-making to assess the quality of decision-making by expert consensus panels. We specifically examine (1) when individual members of panels will divulge their private judgments about the decision to the panel, and (2) when the group judgment is superior to the judgment of individual panelists and will lead to better treatment for patients. We conclude that to maximize the chance of an accurate decision, panels should be made as large as possible, adopt the smallest supermajority rule, and attract members with the highest individual competencies. Furthermore, interdependence among panelists and the goal of reaching consensus can reduce the efficacy of these panels. PMID- 15120471 TI - An economic analysis of adult obesity: results from the Behavioral Risk Factor Surveillance System. AB - This paper examines the factors that may be responsible for the 50% increase in the number of obese adults in the US since the late 1970s. We employ the 1984 1999 Behavioral Risk Factor Surveillance System, augmented with state level measures pertaining to the per capita number of fast-food and full-service restaurants, the prices of a meal in each type of restaurant, food consumed at home, cigarettes, and alcohol, and clean indoor air laws. Our main results are that these variables have the expected effects on obesity and explain a substantial amount of its trend. PMID- 15120472 TI - On the international stability of health care expenditure functions: are government and private functions similar? AB - This paper studies the stability of health care expenditure functions in a sample of OECD countries. We adopt the cointegration approach and the results show that there is a long-term relationship between total health care expenditure (HCE) and gross domestic product (GDP). However, the existence of cointegration is only shown when we admit the presence of some changes in the elasticities of the model. Our results also provide evidence against the existence of a unique relationship between health and GDP for the sample. Thus, we can conclude that the differences in health systems may cause differences in the aggregate functions. Additionally, we examine aggregate health functions for government (GHCE) and private expenditures (PHCE), again finding evidence of different patterns of behaviour. Finally, we open a discussion on the character of health as a necessary or luxury good. In this context, we find differences between the government and the private function. In order to illustrate these findings, we propose a theoretical model as an example of the influence of political decisions on income elasticity. PMID- 15120473 TI - On estimators of medical costs with censored data. AB - In the assessment of cost-effectiveness of alternative medical technologies, it is necessary to estimate the mean total cost per patient over the relevant patient population. Where information about costs comes from a clinical trial with censored data, care is needed to estimate mean total costs. We examine the theoretical connections between the two most widely used of a growing range of nonparametric estimators of costs under censoring. By clarifying the relationships between these simple methods we hope to make them more accessible and to facilitate the take-up of more sophisticated techniques. Recommendations are offered regarding the most appropriate of the available methods, but also on the potential for greater efficiency through parametric modelling. PMID- 15120474 TI - Typical Danish Caucasian type 2 diabetic patients do not commonly carry genetic variants in GIP and GLP-1 encoding regions of the proGIP and proglucagon genes. AB - BACKGROUND: The enteroinsular-axis is abnormal in type 2 diabetics, which contributes to the diabetic phenotype. The effect of the incretin hormone gastric inhibitory polypeptide (GIP) and the secretion of the incretin hormone glucagon like peptide-1 (GLP-1) are thus greatly diminished. The explanation for these changes could be changes in the structure of either of the hormones or their receptors. Thus, the aim of this study was to study the occurrence of genetic variants in the GIP and GLP-1 encoding regions of the proGIP and proglucagon genes in type 2 diabetic patients and matched control subjects. METHODS AND RESULTS: Genomic DNA was extracted from buffy coats from 12 Caucasian type 2 diabetics and 12 healthy subjects, matched with respect to sex, age and BMI. The GIP and GLP-1 sequences were amplified using specific primers using the polymerase chain reaction (PCR). The amplified products were then sequenced. No germ-line mutations were identified in the GIP and the GLP-1 encoding regions of the proGIP and proglucagon genes in either the type 2 diabetic or the control subjects. CONCLUSIONS: The perturbed incretin effect in type 2 diabetics is not commonly caused by genetic variants in either the GIP or the GLP-1 encoding genes in type 2 diabetics. PMID- 15120475 TI - Role of calcitonin gene-related peptide in the phenol-induced neurogenic hypertension in rats. AB - Previous investigations have demonstrated that capsaicin-sensitive sensory nerves are involved in the development of hypertension in some rat models of hypertension. To determine the role played by calcitonin gene-related peptide (CGRP; the predominant neurotransmitter in capsaicin-sensitive sensory nerves) in a rat model of neurogenic hypertension, in which hypertension was induced by injecting 50 microl of 10% phenol in the lower pole of the left kidney, systolic blood pressure (SBP) was monitored by the tail-cuff method throughout the experiment. Fifteen days after injection of phenol, mean arterial pressure (MAP), concentrations of CGRP in the plasma, the expression of CGRP mRNA in dorsal root ganglia (DRG) and CGRP content in laminae I and II of the spinal cord were measured. SBP was significantly increased 5 days after the intrarenal injection of phenol (164+/-7 mm Hg, p<0.01). At the end of experiment, blood pressure (BP) was significantly elevated in the phenol-injected rats compared with the controls (SBP: 187+/-6 vs. 122+/-4 mm Hg, p<0.01; MAP: 157.56+/-3.02 vs. 103.80+/-2.04 mm Hg, p<0.01). Treatment with capsaicin, which selectively depletes neurotransmitters from the capsaicin-sensitive nerves, failed to enhance the development of hypertensive responses to the intrarenal injection of phenol. Intravenous administration of CGRP(8-37), the specific CGRP receptor antagonist, also failed to increase the already elevated MAP. The expression of CGRP mRNA (both alpha- and beta-CGRP isoforms), the content of CGRP in laminae I and II of the dorsal horn of the spinal cord and the concentration of CGRP in the plasma was decreased in the rats treated with phenol. These results suggest that CGRP does not play a counterregulatory role in the phenol-induced hypertensive rats, and support the hypothesis that reduction of CGRP (alpha and beta isoforms) could contribute to a blood pressure elevation in this setting. PMID- 15120476 TI - Functional interaction between transforming growth factor alpha and capsaicin sensitive sensory neurons in the rat stomach. AB - BACKGROUND AND AIMS: Transforming growth alpha (TGFalpha) and sensory neurons have been shown to promote gastric mucosal protection and healing. Aims were to examine in vitro interactions between gastric sensory neurons, the sensory neuropeptide calcitonin gene-related peptide (CGRP), and TGFalpha. METHODS: Gastric mucosal/submucosal tissue fragments from Sprague-Dawley (SD) rats were incubated in short-term (30 min) culture. Peptide release into media and TGFalpha tissue content were measured by radioimmunoassay. RESULTS: TGFalpha (1 x 10(-8) to 1 x 10(-6) M) caused dose-dependent stimulation of CGRP release. Maximal CGRP release (+87%) was observed with 1 x 10(-6) M TGFalpha: 28.6+/-3.8 vs. control of 15.5+/-2.7 pg/g tissue; P<0.05. Both CGRP (1 x 10(-7) to 1 x 10(-5) M) and capsaicin (1 x 10-(8) to 1 x 10(-6)M) significantly inhibited basal TGFalpha release in a dose-dependent fashion that ranged from -20% to -39%. In contrast, capsaicin-induced sensory denervation caused significant increases in both basal TGFalpha release and TGFalpha tissue content. CONCLUSION: Function interactions between TGFalpha and gastric sensory neurons are suggested by the observations that (1) TGFalpha stimulated CGRP release from gastric sensory neurons; (2) CGRP and acute capsaicin treatment inhibited TGFalpha release and; (3) capsaicin induced sensory denervation caused significant increases in both gastric TGFalpha basal release and tissue content. PMID- 15120477 TI - The role of luminal gastrin in the regulation of pancreatic juice secretion in preruminant calves. AB - The effect of luminal gastrin on the secretion of pancreatic juice was studied in seven conscious preruminant calves employing luminal infusions of gastrin and cholecystokinin (CCK)-9 and pharmacological CCK1 and CCK2 receptor blocks with antagonists. The study was performed in the preprandial and prandial states. Pharmacological blocking of the CCK2 receptor, like that of the CCK1 receptor, resulted in reduction of pancreatic postprandial secretion and increased the duration of the prandial pattern of duodenal electrical activity. Exogenous luminal gastrin, like luminal CCK-9, enhanced the secretion of pancreatic juice proteins, though the overall effect of gastrin was weaker than that of CCK-9. The effect was inhibited by infusion of CCK2 but also by CCK1 receptor antagonist. In conclusion, duodenal luminal gastrin can stimulate exocrine pancreatic secretion by a mechanism that depends on CCK2 receptors in calves. Involvement of the CCK1 receptor in this mechanism needs further investigation. Prandial pancreatic secretory and duodenal motility cycles can be regulated by endogenous gastrin release. PMID- 15120478 TI - Differential regulation of central vasopressin receptors in transgenic rats with low brain angiotensinogen. AB - The consequences of permanent alteration to the brain renin-angiotensin system (RAS) on central vasopressinergic system was studied in transgenic rats with low brain angiotensinogen [TGR(ASrAOGEN)]. Levels of vasopressin (AVP) and V1a receptor mRNAs were measured by ribonuclease protection assay (RPA) and AVP by radioimmunoassay (RIA). AVP (100 pmol/50 nl) was microinjected into the nucleus tractus solitarii (NTS) of urethane-anesthetized TGR(ASrAOGEN) and Sprague-Dawley (SD) rats and the mean arterial pressure (MAP) and heart rate (HR) baroreflex induced by phenylephrine were evaluated. AVP but not its mRNA levels were significantly lower in the hypothalamus and hypophysis of TGR(ASrAOGEN) rats. Brainstem V1a mRNA levels were significantly higher in TGR(ASrAOGEN) in comparison to SD rats (5.2+/-0.4% vs. 3.3+/-0.2% of beta-actin mRNA, P<0.05). In contrast, the hypothalamic V1a mRNA levels in TGR(ASrAOGEN) were not different from those found in SD rats. AVP microinjections induced a greater decrease in MAP in TGR(ASrAOGEN) in comparison with SD rats (-19.9+/-5.2 vs. -7.5+/-0.7 mm Hg, P<0.01). The significantly higher baroreflex sensitivity observed in TGR compared to that of SD rats was normalized after AVP microinjection. The increased brainstem V1a mRNA levels and sensitivity to AVP in TGR(ASrAOGEN) rats indicates a functional upregulation of AVP receptors in the NTS. The fact that the hypothalamic V1a mRNA levels are not altered indicates that these receptors are differentially regulated in different brain regions. This study demonstrates that a permanent deficit in brain angiotensinogen synthesis can alter the functionality of central vasopressinergic system. PMID- 15120479 TI - Mutagenesis of the AT1 receptor reveals different binding modes of angiotensin II and [Sar1]-angiotensin II. AB - Homology modeling of the structure of the AT1 receptor, based on the high resolution rhodopsin crystal structure, indicated that it is unlikely that the binding of AngII to AT1 involves simultaneously all the receptor's residues reported in the literature to participate in this process. Site-directed mutagenesis using Ala substitution of charged residues Lys20, Arg23, Glu91 and Arg93 was performed to evaluate the participation of their side-chains in ligand binding and in triggering the cell's response. A comparative analysis by competition binding and functional assays using angiotensin II and the analog [Sar1]-angiotensin II suggests an important role for Arg23 of AT1 receptor in binding of the natural agonist. It is discussed whether some receptor's residues participate directly in the binding with AngII or whether they are part of a regulatory site. PMID- 15120480 TI - Functional analysis of crustacean Hyperglycemic Hormone by in vivo assay with wild-type and mutant recombinant proteins. AB - The neuro-endocrine X-organ sinus-gland complex regulates important crustacean physiological processes, such as growth, reproduction and molting. Its major products are the neuropeptides of the cHH/MIH/GIH family. Until now the structure function relationships of these neuropeptides were established by sequence comparison. To study the functional relevance of conserved amino acid residues or peptide motifs, we generated point and deletion mutants of the Norway lobster Nephrops norvegicus cHH. The wild type mature neuropeptide cHH and its mutant forms were expressed in bacteria as fusion proteins and assayed in vivo to assess their hyperglycemic activity. The wild type cHH had a hyperglycemic activity similar to that of cHH present in an eyestalk extract, and it was blocked by an anti-recombinant cHH antibody. Bioassays of cHHs, obtained by a progressive deletion of five highly conserved motifs, showed that the only deleted cHH, which conserves a hyperglycemic activity, is the one lacking the C-terminal motif, but still retaining all the motifs reported to be important for functional specificity and three-dimensional structure. All the cHH point mutants lacked a hyperglycemic activity. These results identify amino acid residues that are required for the hyperglycemic activity of cHH. PMID- 15120481 TI - Effects of a chromogranin-derived peptide (CgA 47-66) in the writhing nociceptive response induced by acetic acid in rats. AB - Chromogranin A (CgA) is an acidic protein identified within a large variety of endocrine cells. Colocalized with catecholamines in chromaffin cells, CgA is a prohormone precursor of small biologically active peptides. Vasostatin (CgA 1-76) is the most conserved fragment of CgA and chromogranin A 47-66 peptide (CgA 47 66) possesses potent antimicrobial activities. The aim of this study was to test the hypothesis that CgA 47-66 may be involved in mechanisms modulating nociception. Thus, we used acetic acid (AA) which produces a delayed inflammatory response and episodes of abdominal writhing, a marker of pain, when injected intraperitoneally (i.p.) to rats. Administration (i.p.) of CgA 47-66 induced specific opposite dose-dependent effects depending on concentration. That is, CgA 47-66 below 0.5 mg/kg produced antinociceptive effects, whereas at 2 mg/kg it produced a marked pronociceptive effect. The latter effect was blocked by diltiazem and indomethacin. CgA 47-66-induced antinociceptive effects on AA induced responses were reversed when the corticotropin-releasing factor (CRF) antagonist alpha-helical CRF 9-41 was i.p. injected to animals prior to AA and CgA 47-66 administration. The administration of i.p. calcitonin gene-related peptide (CGRP) or substance P (SP) evoked dose-dependent abdominal writhing; this effect was abolished when CgA 47-66 was injected. The present data suggest, for the first time, that a fragment of CgA, CgA 47-66, possesses potent antinociceptive effects at low doses. Although the mechanism triggered by this peptide is unknown, CRF receptors are likely to be involved. PMID- 15120482 TI - Pharmacokinetic profile of orexin A and effects on plasma insulin and glucagon in the rat. AB - Orexin A (OXA) is found in the central nervous system (CNS) and in the gut. Peripheral administration of OXA to rats results in an inhibition of fasting motility. Plasma OXA increases during fasting and central administration of OXA increases food intake. The aim of the present study was to assess the pharmacokinetic profile of OXA and the effect of intravenously (i.v.) administered OXA on plasma concentrations of insulin and glucagon concentrations. Rats were given OXA i.v. (100 pmol kg(-1) min(-1)) for time periods of 0, 10, 20, 30 min and for 10, 20, 30 min after ceasing a 30-min infusion. After each time period, rats were then sacrificed and blood obtained. OXA was also administered at increasing doses (0, 100, 300 and 500 pmol kg(-1) min(-1)) for 30 min and blood was obtained. Plasma OXA, insulin and glucagon levels were measured using commercially available radioimmunoassay (RIA) kits. The plasma half-life of OXA was 27.1+/-9.5 min. Stepwise increasing infusion rates of OXA confirmed a linear concentration-time curve and thus first-order kinetics. Its volume of distribution indicated no binding to peripheral tissues. Plasma glucagon decreased during infusion of OXA, while insulin was unaffected. Plasma OXA was raised fourfold after food intake. Thus, OXA has a longer plasma half-life than many other peptides found in the gut. This needs to be taken into account when assessing effects of OXA on biological parameters after peripheral administration. PMID- 15120483 TI - The role of the pancreatic renin-angiotensin system in acinar digestive enzyme secretion and in acute pancreatitis. AB - The pancreas contains a local renin-angiotensin system (RAS), which is subject to activation by experimental pancreatitis. In the exocrine pancreas, angiotensin II receptor subtypes AT1 and AT2 have been localized in the pancreatic ducts, blood vessels and acinar cells. We hypothesize that local RAS activities may have a potential role in regulating pancreatic acinar digestive enzyme secretion. The present study was designed to elucidate firstly the existence of RAS components in pancreatic acinar cells and their regulation by acute pancreatitis. Secondly, the differential roles of AT1 and AT2 receptors in controlling digestive enzyme secretion from dispersed functional pancreatic acini were also investigated. The mRNA levels of RAS components were assessed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Acinar secretions were assayed by the measurement of alpha-amylase and lipase activities. Induction of acute pancreatitis was achieved by hyperstimulation of two intraperitoneal (i.p.) injections of cerulein (50 microg/kg/h). Results from RT-PCR showed that the mRNA levels of the major RAS components (angiotensinogen, AT1 and AT2 receptors) were expressed in isolated rat pancreatic acinar cells, and they were upregulated during pancreatitis. Exogenous addition of angiotensin II could stimulate a dose dependent release of digestive enzymes from the acinar cells. Administration of the selective AT1 receptor antagonist losartan significantly inhibited the acinar digestion enzyme secretion in both normal and pancreatitis-induced acini. However, a specific AT2 receptor blocker PD123319 did not exhibit such a suppressive effect. These data indicate the existence of an acinar RAS in the pancreas of potential importance in the physiological regulation of digestive enzyme secretion. The differential actions of AT1 and AT2 receptors and their upregulation may have clinical relevance to the pathogenesis and management of acute pancreatitis. PMID- 15120484 TI - Regulation of hypoxia-induced release of corticotropin-releasing factor in the rat hypothalamus by norepinephrine. AB - Corticotropin-releasing factor (CRF) peptide release was activated by hypoxia in the rat hypothalamus. The mechanisms, however, of the hypoxia-induced CRF release remains unclear. In this study, we demonstrated that the norepinephrine (NE) and its receptors in the paraventricular nucleus (PVN) mediated the CRF release in a simulated altitude hypoxia. When rats were exposed to 5 or 7 km altitude of hypoxia for a short or long term: (1) NE levels in the PVN and the CeA, using the HPLC analysis, were intensity and time course dependently increased, but the increase in the PVN were potential than in the CeA. Restraint-induced NE increase was much higher in both the PVN and the CeA, compared with hypoxia-induced response. (2) Hypoxia and restraint significantly enhanced CRF release in the ME and the PVN but not in the CeA, through RIA assay, which result in stimulating corticosterone secretion. (3) Hypoxia-induced CRF release was reversed by an injection of prazosin (i.c.v.), an alpha-1 adrenoceptor antagonist, while administration of yohimbine (i.c.v.), an alpha-2 receptor antagonist, facilitated further CRF release. These data suggested that hypoxia induced NE activation centrally, via alpha-1 and -2 receptors, leading to improving hypothalamic CRF release, which in turn stimulated pituitary and adrenal cortex. Restraint presented much potential action on NE activation than hypoxia. PMID- 15120485 TI - Significance of reductases in the detoxification of the tobacco-specific carcinogen NNK. AB - Fewer than 20% of habitual smokers develop lung cancer, which suggests that genetic, environmental and nutritional factors contribute to the risk for developing this disease. Recently, five enzymes were shown to initiate the detoxification of nicotine-derived nitrosamine ketone (NNK), the most potent carcinogen present in tobacco. Importantly, four of these enzymes are potently inhibited by glycyrrhetinic acid, the main constituent of licorice. These observations might open novel and hitherto unexplored avenues for the risk assessment and prevention of tobacco-associated lung cancer. PMID- 15120486 TI - The inflow and outflow of anti-glaucoma drugs. AB - The treatment of glaucoma by reducing intraocular pressure has relied traditionally on inhibiting aqueous humor secretion (inflow). However, recent therapeutic approaches have targeted aqueous humor outflow. Prostanoid FP receptor agonists selectively increase uveoscleral outflow and the prostamide analog bimatoprost alters trabecular and uveoscleral outflow. An emerging therapeutic strategy is direct neuroprotection of retinal ganglion cells, which are selectively lost in glaucoma. PMID- 15120487 TI - Neuropeptides and anxiety disorders: bombesin receptors as novel therapeutic targets. PMID- 15120489 TI - Reassessing the Th2 cytokine basis of asthma. AB - T helper (Th) 2 cytokines, particularly interleukin 4 (IL-4), IL-5 and IL-13, might be important in the development of allergic asthma. Humanized monoclonal antibodies (hMAbs) against IL-5, and a recombinant soluble human IL-4 receptor have been developed as possible treatments for this disorder. However, these approaches have not yet proven to be successful in the treatment of persistent asthma, suggesting that neither IL-4 nor IL-5 is important in asthma pathogenesis. Indeed, there is insufficient information about the efficacy of soluble IL-4 receptor and the anti-IL-5 hMAbs in the treatment of asthma to draw firm conclusions about the importance of these Th2 cytokines. Nevertheless, because IL-4 is required for IgE production and IL-5 is required for eosinophilopoesis, these Th2 cytokines must remain important candidates for a role in the pathogenesis of allergic asthma. PMID- 15120490 TI - Neuroprotective strategies for Parkinson's disease: conceptual limits of animal models and clinical trials. AB - Parkinson's disease (PD) is a progressive neurodegenerative disorder. Although therapies that treat the symptoms of the disease have proven efficacy, strategies that slow or stop the neurodegenerative process are currently not available. Recently, the National Institute of Neurological Disorders and Stroke (NINDS) conducted a systematic assessment of candidate pharmacological agents with putative neuroprotective properties. Twelve agents have been selected as potential candidates for upcoming clinical trials. However, the data resulting from the use of these agents in animal models of PD using a clinically driven design have not been published. Furthermore, the selection of interesting candidates should be based on the soundest clinically driven preclinical validation. This lack of published data, associated with the conceptual limits of the current way of testing drugs in clinical trials, prompts us to argue for further preclinical validation of the 12 candidates. PMID- 15120491 TI - The potential of GM-CSF to improve resistance against infections in organ transplantation. AB - Immunosuppressed patients retain transplants but become more susceptible to opportunistic infections, which is a major complication in organ transplantation. Life-long immunosuppression for such patients could be reduced by creating immune tolerance, although this might be associated with an increased risk for infections and malignancies. An alternative therapeutic concept could consist of boosting the innate immune response against infections while continuing to suppress the adaptive immune response to prevent graft rejection. We propose granulocyte-macrophage colony-stimulating factor (GM-CSF) as a novel candidate to achieve this goal, based on recent studies in which beneficial effects were demonstrated in immunosuppressed mice with skin allografts and in dexamethasone suppressed blood from healthy volunteers and blood from liver transplant recipients undergoing immunosuppressive therapy. Such data suggest that GM-CSF or other endogenous factors with similar properties should be examined in clinical trials. PMID- 15120492 TI - Nuclear and mitochondrial conversations in cell death: PARP-1 and AIF signaling. AB - Different cell-death mechanisms control many physiological and pathological processes in humans. Mitochondria play important roles in cell death through the release of pro-apoptotic factors such as cytochrome c and apoptosis-inducing factor (AIF), which activate caspase-dependent and caspase-independent cell death, respectively. Poly(ADP-ribose) polymerase 1 (PARP-1) is emerging as an important activator of caspase-independent cell death. PARP-1 generates the majority of long, branched poly(ADP-ribose) (PAR) polymers following DNA damage. Overactivation of PARP-1 initiates a nuclear signal that propagates to mitochondria and triggers the release of AIF. AIF then shuttles from mitochondria to the nucleus and induces peripheral chromatin condensation, large-scale fragmentation of DNA and, ultimately, cytotoxicity. Identification of the pro death and pro-survival signals in the PARP-1-mediated cell-death program might provide novel therapeutic targets in human diseases. PMID- 15120493 TI - The ups and downs of addiction: role of metabotropic glutamate receptors. AB - Drug addiction is characterized by drug-induced positive affect, followed by withdrawal-associated negative affect. Such drug-induced positive and negative affective states provide crucial sources of motivation that drive compulsive drug consumption. Metabotropic glutamate (mGlu) receptors, which are responsible for slow glutamate-mediated neurotransmission, are located throughout limbic and cortical brain regions that are implicated in drug addiction. Emerging evidence indicates that mGlu receptors regulate many behavioral actions of addictive drugs. In particular, group I mGlu receptors play an important role in regulating the reinforcing effects of drugs of abuse. Furthermore, group II mGlu receptors have been implicated in the synaptic adaptations that occur in response to chronic drug exposure and contribute to the aversive behavioral syndrome observed during withdrawal. These findings increase our understanding of the pathological processes that are associated with the development of drug addiction, and might ultimately lead to new therapies for the treatment of this disorder. PMID- 15120494 TI - Sex and estrogen influence drug abuse. AB - Evidence is accumulating that the etiology, epidemiology, consequences and mechanisms that underlie drug abuse are different in males and females. In this review, we present examples of sex differences in all phases of drug abuse, including acquisition, steady-state maintenance, escalation, dysregulation, withdrawal, relapse and treatment. Most reported findings are based on laboratory research in animals, but there are corroborating reports from human clinical and epidemiological studies. In all phases of drug abuse, females seem to be more sensitive to the rewarding effects of drugs than males, and estrogen is a major factor that underlies these sex differences. PMID- 15120495 TI - K+ channels as targets for specific immunomodulation. AB - The voltage-gated Kv1.3 channel and the Ca(2+)-activated IKCa1 K(+) channel are expressed in T cells in a distinct pattern that depends on the state of lymphocyte activation and differentiation. The channel phenotype changes during the progression from the resting to the activated cell state and from naive to effector memory cells, affording promise for specific immunomodulatory actions of K(+) channel blockers. In this article, we review the functional roles of these channels in both naive cells and memory cells, describe the development of selective inhibitors of Kv1.3 and IKCa1 channels, and provide a rationale for the potential therapeutic use of these inhibitors in immunological disorders. PMID- 15120496 TI - Micromorphological analysis of the interaction between a one-bottle adhesive and mineralized primary dentine after superficial deproteination. AB - It has been claimed that resin monomers may incompletely penetrate into demineralized collagen network, which could form a weak hybrid layer. In consequence, it has been proposed that removal of the exposed collagen network could improve adhesion to dentine. The interface between a water/ethanol-based one-bottle adhesive (Single Bond, 3M) which is devoid of acid monomers, and deproteinated surface of primary dentine was evaluated by SEM. Dentine disks were obtained from 20 primary teeth. Two disks were used to standardize the application time of sodium hypochlorite (NaOCl) for getting an effective deproteination. The remaining 18 disks were equally divided into two groups and treated as follows: control group (CG) 35% phosphoric acid (PA) for 15s; treated group/deproteination (TG) 35% PA for 15s+10% NaOCl for 3 min. Single Bond and Z250 (3M) were placed on all disks according to the manufacturer's instructions. The 18 resin-dentine disks were fractured to obtain hemi-disks and processed for SEM. The examination of the CG specimens showed a typical hybrid layer and the presence of numerous tags with few and short microtags. The TG specimens, which did not present hybrid layer, also exhibited numerous tags, with few and short microtags. Some areas between the tags showed fibrillar-like projections, which appeared to be mineralized collagen fibrils, which were incorporated into the adhesive. Thus, our results suggest that some chemical interaction may occur between mineralized dentinal collagen and the adhesive used. PMID- 15120497 TI - Coating of an apatite layer on polyamide films containing sulfonic groups by a biomimetic process. AB - Coating organic polymers with hydroxyapatite is an attractive method for the development of materials for medical applications, as it allows hydroxyapatite to show its unique biological properties such as its ability for bone bonding and protein adsorption. The biomimetic process focuses attention on fabricating such hydroxyapatite-polymer hybrids, where bone-like apatite is deposited on an organic polymer surface in solutions mimicking physiological conditions. In this process, a bone-like apatite layer can be coated onto organic substrates either by using a simulated body fluid (SBF), which has ion concentrations nearly equal to those of human extracellular fluid, or by using fluids that are supersaturated with respect to apatite at ambient conditions. We previously reported that apatite was deposited on polyamide films containing carboxyl groups in a solution mimicking body fluid, when they were incorporated with calcium salts. In the present study, to find an alternative functional group effective in apatite formation, we examined the apatite-forming ability of polyamide films containing sulfonic groups in the same solution. It was found that the polyamide film containing sulfonic groups could deposit apatite on its surface in the solution when the film was incorporated with calcium salts. These results show that the sulfonic group also acts as a functional group, and is as effective for apatite deposition in the body environment as the carboxyl group. PMID- 15120498 TI - Influence of fluoridated mouthwashes on corrosion resistance of orthodontics wires. AB - The aim of the present study was to classify the different alloys commonly used to make orthodontic wire according to their corrosion resistance in different media. The four materials analysed were titanium-based alloys: TMA, TiNb, NiTi and CuNiTi, which were tested in three fluoride mouthwashes: Elmex, Meridol and Acorea as well as in Fusayama Meyer artificial saliva. The electrochemical study showed that the alloys could be divided into two groups. In one group were the NiTi-based alloys which were subject to strong corrosion in the presence of monofluorophosphate found in Acorea solution. In the other group were TiNb, which was the most resistant to corrosion, and TMA, which corroded strongly with the stannous fluoride found in Meridol mouthwash. The results obtained in the present study will enable us to provide attending practitioners with advice concerning fluoride mouthwash to recommend, depending on the treatment phase and the alloy used. So we can advise Elmex mouthwash for patients with TMA and NiTi-based orthodontics wires but we suggest Acorea or Meridol mouthwashes for patients with TiNb orthodontics wires. PMID- 15120499 TI - Investigation of recombinant human elastin polypeptides as non-thrombogenic coatings. AB - We investigated the use of a recombinant human elastin polypeptide as a coating on synthetic materials with a view to determining if these polypeptides could improve the blood compatibility of cardiovascular devices such as vascular conduits and arterial/venous catheters. Platelet adhesion and activation were studied in vitro using three commercially available synthetic materials: polyethylene terephthalate (Mylar), a poly(tetrafluoroethylene/ethylene) copolymer (Tefzel) and a polycarbonate polyurethane (Corethane). Coated with adsorbed polypeptide, all three synthetic materials demonstrated reduced platelet activation and adhesion in platelet rich plasma in vitro. Compared to non-coated controls, there was a significant decrease (p=0.05) in both platelet microparticle release and P-selectin expression for the polypeptide-coated surfaces. Scanning electron microscopy indicated fewer adhering platelets on coated surfaces compared to non-coated controls. In vivo, in a rabbit model, evaluations of polyurethane catheters coated with the polypeptide showed a marked increase in catheter patency and a significant decrease in fibrin accretion and embolism when compared to uncoated controls. This polypeptide shows a strong potential for use as a non-thrombogenic coating for small diameter vascular grafts. In addition, the results of this study indicate that the elastin polypeptide would be a valuable component of a tissue engineered vascular conduit. PMID- 15120500 TI - Synthesis of metal incorporated low molecular weight polyurethanes from novel aromatic diols, their characterization and bactericidal properties. AB - Low molecular weight polyurethanes with sites for metal complexation were synthesized. The -SO(2) and the -COOH groups were incorporated into the polyurethane chain by the reactions of tri-functional monomers, 4,4' bis(hydroxyphenyl)sulfone and 4,4'-bis(hydroxyphenyl) valeric acid with hexa methylene di-isocyanate (HMDI) and 2,4 toluene di-isocyanate (TDI), respectively. The reaction was monitored from the disappearance of the -OH group and results show that the -NCO groups of the isocyanate reacted with the -OH group preferentially when compared to the other reactive groups. The pristine tri functional monomers themselves formed stable complexes with the metals and so were chosen to be incorporated into the polyurethane chain. These polymers were characterized by IR and NMR spectroscopy. The free -SO(2) and the -COOH groups were used for metal complexation using non-toxic metals like zinc and silver. The anti-bacterial studies conducted on the six polymers in film form showed interesting results as elaborated in the paper. PMID- 15120501 TI - The effect of synthetic polymers on the migration of monocytes through human cervical mucus. AB - The integrity of the vaginal mucosa is critical to protecting women from infection, particularly sexually transmitted diseases. For example, breakdown of the mucosa, including the cell lining and/or mucus layer, due to vaginal infections has been shown to increase the risk of HIV infection. There is considerable interest in the development of new topical microbicides for women; many of these topical agents contain polymers. One potential mechanism for altering the barrier properties of a three-dimensional fibrous gel, such as cervical mucus, to cell penetration is to alter the fiber structure. In previous studies, we have shown that addition of synthetic polymers, such as polyvinyl pyridine (PVP) and polyethylene glycol (PEG), modify the fiber structure and mechanical properties of human cervical mucus. Here we investigated the ability of peripheral blood monocytes to migrate through structurally altered mucus gels. Adding PVP to mucus increased both the average fiber spacing and the rate of random migration of monocytes; addition of PEG to mucus also caused an increased random migration rate, although changes in overall fiber spacing were not obvious. In both cases, the addition of small amounts of polymer to cervical mucus decreased the barrier property of mucus with respect to cell migration. This result raises questions about the safety of polymeric agents as ingredients in topical microbicides. PMID- 15120502 TI - Effects of BisGMA on glutathione metabolism and apoptosis in human gingival fibroblasts in vitro. AB - Aim of this study was to investigate the effects of the resin monomer BisGMA on the glutathione concentration (monobromobimane assay) and apoptosis (Annexin V/PI assay) of cultured primary human gingival fibroblasts. Cells were treated for up to 24h with 0.001-0.25 mM BisGMA to determine growth curves using the DNA stain H33342. Subsequent Annexin V/PI-assays revealed that fibroblasts exposed to concentrations of 0.005-0.01 mM (non-cytotoxic) and 0.05 mM (ED(10) concentration) showed no increase of the share of apoptotic cells compared to non treated controls (5-8%), while 0.1 mM BisGMA (approximately ED(50)-concentration) caused a significant increase of the percentage of apoptotic cells (50%). Simultaneously to the induction of apoptosis, 0.1 and 0.25 mM of BisGMA caused a significant depletion of the intracellular GSH content after 18 h of incubation. Our results indicate that BisGMA at concentrations >0.1 mM causes an extreme depletion of the intracellular GSH pool as well as apoptosis. PMID- 15120503 TI - The effect of substrate molecular mobility on surface induced immune complement activation and blood plasma coagulation. AB - Changing the length of the alkyl ester side chain in poly(alkyl methacrylates) provides a unique opportunity to systematically vary the mobility of the polymer chains, or in other words vary the glass transition temperature (T(g)), without greatly affect the solid surface energy (gamma(s)) of the polymer. A series of poly(alkyl methacrylate) coatings was therefore analysed with regard to the human immune complement (IC) activation and the surface associated blood plasma coagulation cascade (CC) properties. For the IC and CC measurements we used a quartz crystal microbalance (QCM) where we modified the chemistry of the sensor surface by applying 10-30 nm thick poly(alkyl methacrylate) coatings. The surface energy was calculated from water contact angles and small differences between the coatings were observed. The surface chemistry of the coatings, as determined with X-ray photoelectron spectroscopy (XPS), showed no deviation from expected compositions. Tapping mode atomic force microscopy (TM-AFM) measurements revealed that all coatings displayed similar morphology and the roughness was in the range of 0.7-0.9 nm. Increased polymer mobility correlated with a decrease in IC activation, measured as a decreased C3c deposition at the surface. The surface induced CC, measured as fibrin clot formation at the surface, was different between the different coatings but no correlation with molecular mobility was observed. Thus, the molecular mobility of the polymer chains had a major effect on both the IC and the CC and it seems that different aspects of the chemistry of the solid surface regulate activation of the IC and the CC. PMID- 15120504 TI - Soluble eggshell membrane protein: preparation, characterization and biocompatibility. AB - The preparation, characterization and biocompatibility of soluble eggshell membrane (SEP) are reported. The dissolution process, which is the key step of the preparation of SEP, has been followed by scanning electron microscopy (SEM) to observe the changes of the surfaces and thickness of the eggshell membrane (ESM). The composition of SEP has been investigated by amino acid analysis and elemental analysis. Based on the fact that SEP losses significantly cystine, and that SEP has a higher content of sulfur, an assumption involving combination with 3-mercaptopropionic acid (the reagent used for reductive cleavage of disulfide bonds) following the cleavage of the original disulfide bonds has been proposed, which explains the solubility of SEP. The thermal and surface properties have been studied by thermogravimetric analysis (TGA) and contact angle measurement. The biocompatibility of SEP, as demonstrated by cell culture of NIH3T3, is comparable to collagen type I and superior to raw ESM either inside or outside surface. PMID- 15120505 TI - Evaluation of titanium plasma-sprayed and plasma-sprayed hydroxyapatite implants in vivo. AB - In this study, bone interfacial strength and bone contact length at the plasma sprayed hydroxyapatite (HA) and titanium plasma-sprayed (TPS) implants were evaluated in vivo. Non-coated titanium (Ti) implants were used as controls. Cylindrical coated or non-coated implants (4.0mm diameter by 8mm long) were implanted in the dogs' mandibles. Loading of the implants was performed at 12 weeks after implantation. At 12 weeks after implantation (prior to loading) and 1 year after loading, implants were evaluated for interfacial bone-implant strength and bone-implant contact length. No significant differences in interfacial bone implant strength for all groups at 12 weeks after implantation and after 1 year loading in normal bone were found. However, bone contact length for HA implants was significantly higher than the TPS and Ti implants for both periods tested (12 weeks after implantation and 1 year after loading). It was concluded that TPS implants exhibited similar pull-out strength compared to the HA implants. In addition, the lower bone contact length on the TPS surface compared to HA surfaces did not affect the interfacial bone-implant strength for both implants. PMID- 15120506 TI - Laser surface modification of hydroxyapatite and glass-reinforced hydroxyapatite. AB - Surface treatment of materials with excimer laser radiation often results in the formation of a rough columnar or cone-shaped surface topography, which leads to a considerable increase in the surface area. As a result, the search for a non porous bioactive material with adequate mechanical properties and a high surface to volume ratio, similar to porous materials, which could be used for drug delivery in the treatment of periodontitis, justified assessing excimer laser surface treatment to promote controlled roughning of hydroxyapatite (HA) and glass-reinforced hydroxyapatite (GR-HA). A KrF excimer laser with 248 nm radiation wavelength and 30 ns pulse duration was used for surface modification. The laser treatment was carried out in air, using wide ranges of radiation fluence and number of laser pulses. In order to identify the physico-chemical changes induced by the laser treatment and the column formation mechanisms in these materials, the treated surfaces were characterised by laser profilometry, scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and Fourier transform infra-red spectroscopy (FTIR). Laser processing induced the formation of a surface topography consisting of cone shaped features. The constitution of the surface layer was also modified, as revealed by FTIR, XPS and XRD. This work has shown that laser surface modification increases the surface area of HA and GR-HA and is a promising technique to increase the reactivity and drug delivery capability of both materials. PMID- 15120507 TI - Strong and bioactive composites containing nano-silica-fused whiskers for bone repair. AB - Self-hardening calcium phosphate cement (CPC) sets to form hydroxyapatite with high osteoconductivity, but its brittleness and low strength limit its use to only non-stress bearing locations. Previous studies developed bioactive composites containing hydroxyapatite fillers in Bis-GMA-based composites for bone repair applications, and they possessed higher strength values. However, these strengths were still lower than the strength of cortical bone. The aim of this study was to develop strong and bioactive composites by combining CPC fillers with nano-silica-fused whiskers in a resin matrix, and to characterize the mechanical properties and cell response. Silica particles were fused to silicon carbide whiskers to roughen the whisker surfaces for enhanced retention in the matrix. Mass ratios of whisker:CPC of 1:2, 1:1 and 2:1 were incorporated into a Bis-GMA-based resin and hardened by two-part chemical curing. Composite with only CPC fillers without whiskers served as a control. The specimens were tested using three-point flexure and nano-indentation. Composites with whisker:CPC ratios of 2:1 and 1:1 had flexural strengths (mean+/-SD; n=9) of (164+/-14) MPa and (139+/ 22) MPa, respectively, nearly 3 times higher than (54+/-5) MPa of the control containing only CPC fillers (p<0.05). The strength of the new whisker-CPC composites was 3 times higher than the strength achieved in previous studies for conventional bioactive composites containing hydroxyapatite particles in Bis-GMA based resins. The mechanical properties of the CPC-whisker composites nearly matched those of cortical bone and trabecular bone. Osteoblast-like cell adhesion, proliferation and viability were equivalent on the non-whisker control containing only CPC fillers, on the whisker composite at whisker:CPC of 1:1, and on the tissue culture polystyrene control, suggesting that the new CPC-whisker composite was non-cytotoxic. PMID- 15120508 TI - Inhibition of PMN apoptosis after adherence to dip-coated calcium phosphate surfaces on a NiTi shape memory alloy. AB - Nickel-titanium shape-memory alloys (NiTi-SMA) were coated with calcium phosphate by dipping in oversaturated calcium phosphate solution (CaP-coating). Polymorphonuclear neutrophil granulocytes (PMN) belong to the first cells which will adhere to implant materials. We analyzed the apoptosis of isolated human PMN after cell culture on non-coated and CaP-coated NiTi-SMA by light and scanning electron microscopy (cell morphology) and by flow cytometry (DNA-fragmentation). In contrast to PMN adherent to non-coated NiTi-SMA, the apoptosis of PMN adherent CaP-coated samples was inhibited. Cell culture media obtained from cultured leukocytes with CaP-coatings (conditioned media, CM) were able to transfer the apoptosis inhibiting activities to freshly isolated PMN. There was a significant (p<0.01) increase in GM-CSF, IL-8, IL-6, and TNF-alpha within CM obtained from coated versus non-coated NiTi-SMA as was determined by ELISA. PMID- 15120509 TI - The effect of polylactide membranes on the levels of reactive oxygen species in periodontal flaps during wound healing. AB - It is consented that reactive oxygen species (ROS) are deleterious to wound healing process due to the harmful effects on cells and tissues. Absorbable synthetic biomaterials are considered to be degraded via ROS. Free-radical scavenging enzymes (FRSE) are a cytoprotective enzymal group that has an essential role in the reduction, de-activation and removal of ROS as well as regulating wound healing process. In the present study, synthetic and absorbable polylactide (PLA) barrier membranes were evaluated by means of ROS activity levels during degradation in the healing periodontal flaps measuring the activity of FRSE superoxide dismutase (SOD) and catalase (CAT). Gingival biopsies taken from 10 patients allowing both guided tissue regeneration (test) and conventional flap surgery (control) before and 1 month after the operations were processed and the supernatants were studied by Mc Cord and Fridovich, Flohe and Otting, and Luck methods to measure total SOD and CAT levels respectively. A significantly increased enzyme activity of SOD and CAT was observed in both groups (p<0.05). SOD activity change was 62.92% in the test and 3.97% in the control group, and, CAT activity change was 48.04% in the test and 11.58% in the control group. Our results suggest that ROS, particularly superoxide anions, may contribute to the degradation phase of PLA membranes and this may affect the wound healing of periodontium at least for one-month period. PMID- 15120510 TI - The in vivo performance of a sol-gel glass and a glass-ceramic in the treatment of limited bone defects. AB - The in vivo evaluation, in New Zealand rabbits, of a SiO(2)-P(2)O(5)-CaO sol-gel glass and a SiO(2)-P(2)O(5)-CaO-MgO glass-ceramic, both bioactive in Kokubo's simulated body fluid (SBF), is presented. Bone defects, performed in the lateral aspect of distal right femoral epiphysis, 5mm in diameter and 4mm in depth, were filled with (i) sol-gel glass disks, (ii) glass-ceramic disks, or (iii) no material (control group). Each group included 8 mature and 8 immature rabbits. A 4-month radiographic study showed good implant stability without axial deviation of extremities in immature animals and periosteal growth and remodelling around and over the bone defect. After sacrifice, the macroscopic study showed healing of bone defects, with bone coating over the implants. The morphometric study showed a more generous bone formation in animals receiving sol-gel glass or glass ceramic disks than in control group. Histomorphometric study showed an intimate union of the new-formed bone to the implants. This study allows considering both materials as eligible for bone substitution or repair. Their indications could include cavities filling and the coating of implant surfaces. The minimum degradation of glass-ceramic disks suggests its application in locations of load or transmission forces. As specific indication in growth plate surgery, both materials could be used as material of interposition after bony bridges resection. PMID- 15120512 TI - Stability of plasma-treated silicone rubber and its influence on the interfacial aspects of blood compatibility. AB - Medical-grade polydimethylsiloxane elastomer was subjected to low-powered plasma treatment in the presence of four different gases: O(2), Ar, N(2) and NH(3). Changes to the surface chemistry immediately after processing and the stability of the treatments following ageing in phosphate buffered saline or air for up to 1 month were investigated using X-ray photoelectron spectroscopy and dynamic contact angle analysis. Changes in surface morphology were assessed using optical microscopy and atomic force microscopy. All treatments resulted in an increase in wettability, attributed to major changes in chemistry combined with modest etching. Furthermore, the primary site of attack of the plasma species appeared to be dependent upon the feed gas implemented. The two main chemical changes observed after ageing were due to reactions with the storage media and relaxation processes resulting in further changes in wettability. The influence of the surface modifications on the blood compatibility of the materials was investigated by assessing contact phase activation using a partial thromboplastin time assay. It was demonstrated that the O(2) and Ar plasma treatments reduced the performance of the silicone but the N(2) and NH(3) treatments had a significantly beneficial effect on the activation of the coagulation cascade. PMID- 15120511 TI - Preparation of magnesium-substituted hydroxyapatite powders by the mechanochemical-hydrothermal method. AB - Magnesium-substituted hydroxyapatite (Mg-HAp) powders with different crystallinity levels were prepared at room temperature via a heterogeneous reaction between Mg(OH)(2)/Ca(OH)(2) powders and an (NH(4))(2)HPO(4) solution using the mechanochemical-hydrothermal route. The as-prepared products contained unreacted Mg(OH)(2) and therefore had to undergo purification in ammonium citrate aqueous solutions at room temperature. X-ray diffraction, infrared spectroscopy, thermogravimetric and chemical analyses were performed and it was determined that the purified powders were phase-pure Mg-HAp containing 0.24-28.4 wt% of Mg. The concentration of Mg was slightly lower near the surface than in the bulk of the HAp crystals as indicated by X-ray photoelectron spectroscopy. Dynamic light scattering revealed that the median particle size of the room temperature Mg-HAp powders was in the range of 102 nm-1.2 microm with a specific surface area between 91 and 269 m(2)/g. Scanning electron microscopy confirmed that the Mg-HAp powders consisted of submicron agglomerates of nanosized crystals, less than approximately 20 nm. PMID- 15120513 TI - Decrease in particle-induced osteolysis in obese (ob/ob) mice. AB - There may be variability in the susceptibility of different individuals to osteolysis from wear debris, and it is not clear whether some individuals may have a genetic predisposition for a more marked osteolytic response. The purpose of this study in mice was to determine whether genetically determined obesity can alter the response to particulate debris. Polyethylene particles were implanted onto the calvaria of seven wild-type mice and seven obese mice (ob/ob). Calvaria from unimplanted wild-type and obese mice served as controls. Calvaria were harvested after 7 days, stained with toluidine blue and for tartrate-specific alkaline phosphatase, and analyzed by histomorphometry. The osteoclast number per mm total bone perimeter was 8.000+/-3.464 in wild-type animals with particles and 2.857+/-1.676 in ob/ob animals with particles (p=0.002; Fisher's PLSD). Bone resorption was 1.895+/-0.713 mm/mm(2) in wild-type animals with particles and 1.265+/-0.494 mm/mm(2) in ob/ob animals with particles (p=0.0438; Fisher's PLSD). Particles induced a diminished osteolytic response in genetically determined obese mice, suggesting that obesity may have a protective role against particle induced bone resorption-similar to obesity and osteoporosis. These important new findings may help to stimulate clinical studies which may define criteria to better identify patients at risk to develop particle-induced osteolysis. PMID- 15120514 TI - Fabrication of microstructures in photosensitive biodegradable polymers for tissue engineering applications. AB - Combining the MEMS technology and biology requirements for tissue engineering, the fabrication processes of microstructured chambers and microchannels made in biodegradable photosensitive polymers are presented. The fabrication processes, based on softlithography are very fast and flexible. Various single and multistepwise microstructures could be achieved using the biodegradable polymers. Microstructures down to 50microm, which are suitable for liver reconstructs, could be fabricated. As the pCLLA acrylate photosensitive polymer has interesting property for implantable bioreactors, that is, its softness, we examined the ability of various mammalian cells to grow and spread on it. With Hep G2 cells, human umbilical blood vessel endothelial cells (HUVEC), 3T3-L1 mouse fibroblasts, static cultures could be successfully performed on single stepwise microstructures. Then, by using this photosensitive biodegradable polymer, a microstructure with simple fluidic channels is fabricated and a perfusion experiment could be carried out. Both cell cultures and perfusion experiments suggested the possibility to use the present photosensitive polymer as microfluidic supports for biodegradable bioreactors for implantation applications. PMID- 15120515 TI - Basic study of corn protein, zein, as a biomaterial in tissue engineering, surface morphology and biocompatibility. AB - The zein films, were prepared for culturing human liver cells (HL-7702) and mice fibroblast cells (NIH3T3), while the Corning microplate and polylactic acid (PLA) were chosen as controls. The surface morphology of zein films prepared by two different methods was studied by scanning electron microscope (SEM), which revealed that the zein films were composed of particles of diameter 100-500 and 500-2500 nm, respectively. The biocompatibility of zein films was assessed by attachment, extensibility and proliferation of cells on them. Our study indicated that over 60% of both HL-7702 cells and NIH3T3 cells could attach to the Corning microplate, zein films and PLA at 3h after seeding. The concentration and particle sizes for preparing zein films did not seem to affect the proliferation of the cells tested. There were no significant differences in the proliferation of both HL-7702 cells and NIH3T3 cells between the Corning microplate and two kinds of zein films, except that the zein film composed of smaller particles at the lowest concentration exhibited a very good ability for proliferation of both the cells, while PLA was a poor matrix in the latter period of the cell proliferation. This preliminary study demonstrates that zein is a promising biomaterial with good biocompatibility for the development of tissue engineering. PMID- 15120516 TI - Response of MG63 osteoblast-like cells onto polycarbonate membrane surfaces with different micropore sizes. AB - Response of different types of cells on materials is important for the applications of tissue engineering and regenerative medicine. It is recognized that the behavior of the cell adhesion, proliferation, and differentiation on materials depends largely on surface characteristics such as wettability, chemistry, charge, rigidity, and roughness. In this study, we examined the behavior of MG63 osteoblast-like cells cultured on a polycarbonate (PC) membrane surfaces with different micropore sizes (0.2-8.0 microm in diameter). Cell adhesion and proliferation to the PC membrane surfaces were determined by cell counting and MTT assay. The effect of surface micropore on the MG63 cells was evaluated by cell morphology, protein content, and alkaline phosphatase (ALP) specific activity. It seems that the cell adhesion and proliferation were progressively inhibited as the PC membranes had micropores with increasing size, probably due to surface discontinuities produced by track-etched pores. Increasing micropore size of the PC membrane results in improved protein synthesis and ALP specific activity in isolated cells. There was a statistically significant difference (P<0.05) between different micropore sizes. The MG63 cells also maintained their phenotype under conditions that support a round cell shape. RT-PCR analysis further confirmed the osteogenic phenotype of the MG63 cells onto the PC membranes with different micropore sizes. In results, as micropore size is getting larger, cell number is reduced and cell differentiation and matrix production is increased. This study demonstrated that the surface topography plays an important role for phenotypic expression of the MG63 osteoblast-like cells. PMID- 15120517 TI - Combination of porous hydroxyapatite and cationic liposomes as a vector for BMP-2 gene therapy. AB - The clinical significance of hydroxyapatite (HAP) as a bone substitute has become apparent in recent years and bone morphogenetic protein (BMP) a substance which induces bone has attracted much attention. In this study, a 1.2 cm diameter bone defects created on rabbit cranium were treated with the BMP-2 gene (cDNA plasmid) introduced with porous HAP after completion of hemostasis and the resultant bone formation was analyzed histopathologically. The amounts of bone formation was compared BMP-2 cDNA plasmids were not combined with cationic liposomes as a vector. Four groups of rabbits were compared. In the HAP group the cranial bone defect was treated with HAP containing 40 microg of liposomes and a dummy gene (PU). The BMP gene HAP group was treated with HAP soaked in liposomes and 10 microg of the BMP-2 gene. In addition, a group was treated with the gene without implanting HAP. Bone formation on the cranial defects was evaluated 3, 6 and 9 weeks after the operation, by X-ray and histopathological examinations. Three weeks after the operation there was vigorous bone formation in the cranial defect in the group which received the BMP-2 gene without HAP, and complete ossification was observed at 9 weeks. In the group which received HAP containing the BMP-2 gene, although new bone formation was evident surrounding the scaffold 3 weeks post-operation, the induced bone tissue did not fill all the pores of the scaffold even at 9 weeks post-operation. These results confirm the clinical usefulness of gene therapy for bone formation, using the BMP-2 gene combined with cationic liposomes as a vector. It is possible that the effects of administering the BMP-2 gene will be improved by specializing the microstructure of scaffold for gene therapy. PMID- 15120518 TI - Synthesis and characterization of partially biodegradable, temperature and pH sensitive Dex-MA/PNIPAAm hydrogels. AB - The objective of the study is to impart temperature and pH-sensitive capabilities to polysaccharide-based hydrogels, so that they can change their swelling property upon external stimulation like temperature or/and pH. Dextran was chosen as the model polysaccharide compound for such a demonstration. A novel class of dextran-maleic anhydride (Dex-MA)/poly(N-isopropylacrylamide) hybrid hydrogels was designed and synthesized by UV photocrosslinking. The dextran-based precursor (Dex-MA) was prepared by substituting the hydroxyl groups in Dex by MA. This Dex MA precursor was then photocrosslinked with a known temperature sensitive precursor (N-isopropylacrylamide, NIPAAm) to form hybrid hydrogels having a wide range of composition ratio of Dex-MA to NIPAAm precursors. Due to the biodegradable nature of dextran, these Dex-MA/PNIPAAm hybrid hydrogels are partially biodegradable. These smart hybrid hydrogels were characterized by Fourier transform infrared spectroscopy for structural determination, differential scanning calorimertry for thermal property, maximum swelling ratio, swelling kinetics, temperature response kinetics, and effect of pH. The data obtained clearly show that these new smart hybrid hydrogels were responsive to the external changes of temperature as well as pH. The magnitude of smart and hydrogel properties of these hybrid hydrogels were found to depend on the feed composition ratio of the two precursors. By changing the composition ratio of these two precursors, the phase transition temperature (lower critical solution temperature) of the hybrid hydrogels could also be adjusted to be or near the body temperature for the potential applications in bioengineering and biotechnology fields. PMID- 15120519 TI - Increased osteoblast adhesion on nanophase metals: Ti, Ti6Al4V, and CoCrMo. AB - Previous studies have demonstrated increased functions of osteoblasts (bone forming cells) on nanophase compared to conventional ceramics (specifically, alumina, titania, and hydroxyapatite), polymers (such as poly lactic-glycolic acid and polyurethane), carbon nanofibers/nanotubes, and composites thereof. Nanophase materials are unique materials that simulate dimensions of constituent components of bone since they possess particle or grain sizes less than 100 nm. However, to date, interactions of osteoblasts on nanophase compared to conventional metals remain to be elucidated. For this reason, the objective of the present in vitro study was to synthesize, characterize, and evaluate osteoblast adhesion on nanophase metals (specifically, Ti, Ti6Al4V, and CoCrMo alloys). Such metals in conventional form are widely used in orthopedic applications. Results of this study provided the first evidence of increased osteoblast adhesion on nanophase compared to conventional metals. Interestingly, osteoblast adhesion occurred preferentially at surface particle boundaries for both nanophase and conventional metals. Since more particle boundaries are present on the surface of nanophase compared to conventional metals, this may be an explanation for the measured increased osteoblast adhesion. Lastly, material characterization studies revealed that nanometal surfaces possessed similar chemistry and only altered in degree of nanometer surface roughness when compared to their respective conventional counterparts. Because osteoblast adhesion is a necessary prerequisite for subsequent functions (such as deposition of calcium containing mineral), the present study suggests that nanophase metals should be further considered for orthopedic implant applications. PMID- 15120520 TI - Surface properties and biocompatibility of solvent-cast poly[-caprolactone] films. AB - Poly(epsilon-caprolactone) (PCL) was dissolved in four solvent systems, chloroform, tetrahydrofuran, acetone and ethyl acetate, and cast onto glass Petri dishes. The surface properties of the resulting films were investigated. The extent to which their properties were determined by the solvent used in each case was quantified in terms of contact angle, surface morphology, attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR), and the adhesion and proliferation of fibroblasts by direct contact. The surface of the PCL film in contact with glass was denoted the SG surface, and the other, which was exposed to the gas phase, a mixture of air and residual solvent vapour, was denoted the SA surface. In the case of hydrophobic solvent systems, the advancing contact angle of the SG surface was always lower than that of the SA surface. With hydrophilic solvent systems, on the other hand, the advancing contact angle of the SG film surface was higher when the contact angle of the Petri dish was higher than that of the gaseous mixture of the air and solvent vapour, otherwise it was lower or equal to that of the surface on which it was cast. The surface morphology was dictated by the solubility of PCL in the respective solvent systems: high dissolution solvents such as chloroform and tetrahydrofuran produced films that comprised PCL aggregates, the particles being larger in the case of chloroform, whereas the less efficient solvents (acetone and ethyl acetate) resulted in a filamentous structure. The ATR-FTIR results confirmed that the chemistry of the SA surfaces differed according to the solvent system used. Preliminary cell culture experiments carried out with the PCL films established that murine (L929) fibroblasts grew well on all surfaces regardless of the solvent used, although the rates of adhesion and proliferation were not as great as on tissue culture plastic controls. Of all the surfaces examined in this study, the cells favoured the SG aspect of ethyl acetate cast PCL films, the surface of which had the finest pore size and relatively low contact angle. PMID- 15120521 TI - Structure and properties of nano-hydroxyapatite/polymer composite scaffolds for bone tissue engineering. AB - To better mimic the mineral component and the microstructure of natural bone, novel nano-hydroxyapatite (NHAP)/polymer composite scaffolds with high porosity and well-controlled pore architectures were prepared using thermally induced phase separation (TIPS) techniques. The morphologies, mechanical properties and protein adsorption capacities of the composite scaffolds were investigated. The high porosity (90% and above) was easily achieved and the pore size was adjusted by varying phase separation parameters. The NHAP particles were dispersed in the pore walls of the scaffolds and bound to the polymer very well. NHAP/polymer scaffolds prepared using pure solvent system had a regular anisotropic but open 3D pore structure similar to plain polymer scaffolds while micro-hydroxyapatite (MHAP)/polymer scaffolds had a random irregular pore structure. The introduction of HAP greatly increased the mechanical properties and improved the protein adsorption capacity. In a dioxane/water mixture solvent system, NHAP-incorporated poly(L-lactic acid) (PLLA) scaffolds developed a fibrous morphology which in turn increased the protein adsorption three fold over non fibrous scaffolds. The results suggest that the newly developed NHAP/polymer composite scaffolds may serve as an excellent 3D substrate for cell attachment and migration in bone tissue engineering. PMID- 15120522 TI - Implantation of hydrophilic and hydrophobic titanium discs in rat tibia: cellular reactions on the surfaces during the first 3 weeks in bone. AB - In a previous study, a method for evaluation of short-time (1-8 days) healing of titanium implants in rat tibiae was described (J. Biomed. Mater. Res. 66A(3) (2003) 662). The implants were disc-shaped and cells and tissue on the surface were investigated, not the adjacent tissue. In this study healing during the first 3 weeks in bone was examined and the healing response between hydrophilic and hydrophobic titanium was compared. Immunofluorescence techniques were used to detect signs of bone formation on the surfaces. Cell viability, alkaline phosphatase (ALP) activity, presence of osteocalcin and cells positive for bone morphogenetic protein-2 (BMP-2) and vascular endothelial growth factor (VEGF) were investigated. Both viable and non-viable cells were found on both surfaces during the first week. Only initially was there a difference between them; 4% viable cells on hydrophilic discs compared to 56% on hydrophobic ones. More BMP-2 positive cells were found on hydrophilic discs than on hydrophobic ones after 1 week. VEGF was detected after 8 days on both surfaces. Osteocalcin positive cells were found from 2 weeks. ALP positive cells were found after 8 days, while at 2-3 weeks ALP positive tissue was abundant on both surfaces. In conclusion, signs of bone formation were detected during the period investigated. Surface energy appeared to be of more importance initially, with higher surface energy resulting in more rapid cell activation and differentiation than lower. PMID- 15120523 TI - Hot embossing for micropatterned cell substrates. AB - This paper reports the development of a technique for preparing microtextured polymer substrates for cell growth and studies the response of osteoblast cells grown on these surfaces. The surfaces were manufactured with hot embossing, where a silicon micromachined printing master was pressed into a thermoplastic polymer substrate at elevated temperature, forming a regular microgroove pattern in the polymer. The grooves were approximately 5 microm deep, 4 microm wide, and had a periodicity of 34 microm. The polymer substrate was polyimide, which can be spincast and printed in its uncured form, and is mechanically rigid and chemically nonreactive after full cure. Osteoblast cells were grown on the textured polymer substrate and their responses to grooved and smooth surfaces were observed with fluorescence microscopy. Alignment and aspect ratio were analyzed for the cell body, cell nucleus, and focal adhesions. Cell membrane body, cell nucleus, and focal adhesions all strongly aligned with the microgrooves, while only the cell body shape changed on the microgrooved surface. This novel substrate preparation technique offers the opportunity for low-cost and rapid manufacture of microtextured surfaces that can be used to control cell shape and alignment. PMID- 15120524 TI - Characterization of surface property of poly(lactide-co-glycolide) after oxygen plasma treatment. AB - In this study, poly (lactide-co-glycolide) (PLGA) films were treated by oxygen plasma. The surface structure, topography and surface chemistry of treated PLGA films were characterized by contact angle measurement, scanning electron microscope observation, atomic force microscopy, and X-ray photoelectron spectrum analysis. The cell affinity of the oxygen plasma treated films was evaluated under dynamic conditions by Parallel Plate Flow Chamber (PPFC). The results showed that the hydrophilicity increased greatly after oxygen plasma treatment. High quantities of -C-O groups, such as hydroxyl and peroxyl groups could be incorporated into the surface of PLGA (70/30) by controlling appropriate plasma treatment conditions. Moreover, the oxygen plasma treatment resulted in formation of peaks and valleys on the sample surfaces, and the roughness increased with treatment time. Cells stretched very well and the ability to endure the shear stress was improved greatly after the PLGA (70/30) was modified by appropriate plasma treatment, i.e. under 50W for 2 or 10 min. However, when the treatment time was increased to 20 min, the percentage of adherent cells on the roughest surface decreased because the content of polar groups incorporated onto the surface decreased. The results showed that improved cell adhesion was attributed to the combination of surface chemistry and surface morphology of PLGA during plasma etching. PMID- 15120526 TI - Prolongation of sciatic nerve blockade by in situ cross-linked hyaluronic acid. AB - Controlled release technology has been applied extensively in providing prolonged duration local anesthesia. Here we used modified hyaluronic acids (HAs; hydrazide and aldehyde) that cross-link upon mixing, as the vehicle for bupivacaine. We assessed the formulations' efficacy and biocompatibility in a rat model of sciatic nerve blockade. We found that 2% (w/v) cross-linked HA doubled the duration of block of 0.1%, 0.25%, and 0.5% (w/v) bupivacaine, without a statistically significant increase in myotoxicity. 1% (w/w) cross-linked HA also prolonged nerve block, but unmodified HA, and both modified HAs did not. HA itself was associated with a mild to moderate inflammatory response with macrophages and lymphocytes. Cross-linked HA is an effective and biocompatible vehicle for enhancing local anesthetic efficacy. PMID- 15120525 TI - Biological evaluation and drug delivery application of cationically modified phospholipid polymers. AB - Phospholipid-like polymers based on 2-methacryloyloxyethyl phosphorylcholine containing varying amounts of the cationically charged monomer choline methacrylate (CMA) from 0 to 30 wt% have been prepared. Substrates coated with these materials were shown to bind significantly lower amounts of specific proteins compared to the uncoated control. ELISA assays demonstrated that fibrinogen did not bind appreciably to coatings containing 0-30% CMA, whereas albumin binding was seen to increase significantly as the CMA content of the coating increased. Platelet activation assays, measurement of plasma coagulation time and whole blood contact scanning electron micrography demonstrated that the haemocompatibility of the coatings was shown to be unaffected by the CMA component. The CMA polymer coatings have been shown to absorb/adsorb many different drug compounds covering a wide range of molecular weights and release these in a controlled fashion. The range of cationic polymers assessed can interact with the net negative charge found in many large therapeutic biomolecules, such as DNA fragments used in gene therapy, that may be of interest in the preventative treatment of conditions such as restenosis. Coronary stents coated with 6% or 20% CMA-containing polymers have been shown to load and release this type of genetic material irrespective of molecular weight of the biomolecule. Ex vivo and in vivo studies have shown that these compounds can be delivered to the stented section of the vessel with very low quantities delivered outside the vessel target area. PMID- 15120527 TI - Magneto-mechanical stimulation of bone growth in a bonded array of ferromagnetic fibres. AB - A brief experimental and theoretical study is presented into the elastic deformation of bonded arrays of ferromagnetic fibres, when subjected to an external magnetic field. Material made of such fibre arrays is of potential interest for certain biomedical applications, such as prosthetic implants. Externally imposed magnetic fields could be used to generate mechanical strains in surrounding tissue, with possible physiological benefits. It is shown that it should be possible to generate strains within embryonic bone cell networks, forming within such a fibre array, which are sufficient to stimulate enhanced growth. The effects outlined here could thus form the basis of surgical or therapeutic advances. PMID- 15120528 TI - Surface modification of poly(ethylene-co-vinyl alcohol): hydroxyapatite immobilization and control of periodontal ligament cells differentiation. AB - To reveal and control the differentiation and proliferation of the periodontal ligament (PDL) cells and to develop a highly organized hybrid implant possessing periodontium, hydroxyapatite (HAP) was immobilized on the poly(ethylene-co-vinyl alcohol) (EVA) by alternate soaking method following with carboxyl groups' introduction through ozone exposure. Human PDL cells were cultured on the ozone exposed EVA, collagen-immobilized EVA, HAP-immobilized EVA, HAP plate, tricalcium phosphate plate, and conventional tissue culture dish. Cell proliferation was highest on the collagen-immobilized EVA and lowest on the HAP-immobilized EVA. Alkaline phosphatase activity and osteocalcin secretion were highest on the HAP immobilized EVA. These results suggest that PDL cells were differentiated toward bone-like cells on the HAP-immobilized EVA. PMID- 15120529 TI - Presence of fibrinogen-binding adhesin gene in Staphylococcus epidermidis isolates from central venous catheters-associated and orthopaedic implant associated infections. AB - Attention has recently been paid to identify and elucidate those pathogenetic mechanisms, which play a significant role in sustaining the early phases of Staphylococcus epidermidis colonisation and infection development. Several analogies with the physiology of Staphylococcus aureus, a more thoroughly investigated pathogen, have lead to carefully consider all bacterial surface components that mediate cell adhesion. This study aimed at investigating the presence of the fbe gene encoding for a fibrinogen-binding protein in a collection of 107 S. epidermidis strains isolated from orthopaedic infections and 67 from central venous catheter-associated infections. The strains isolated from orthopaedic infections were in large part associated to four different classes of orthopaedic devices, respectively: internal fixation devices, external fixation devices, knee arthroprostheses and hip arthroprostheses. The molecular epidemiology analysis performed by PCR enlightened a statistically significant difference in the prevalence of this adhesion mechanism between orthopaedic infections and catheter-related infections, respectively, of 78% and 91%. The prevalence of fbe ranged from 67% to 91%, suggesting that, even though this adhesin is not strictly necessary for the development of infection, nevertheless it represents a rather common characteristic of strains causing clinical infections, this independently on the presence or the absence of implant materials. PMID- 15120530 TI - Comparison of cell viability on anorganic bone matrix with or without P-15 cell binding peptide. AB - Cell-extracellular cell matrix (ECM) interactions play a vital role in development including cytoskeletal structure, growth and differentiation. A 15 amino acid, non-RGD containing, peptide that is derived from type I collagen was tested for its ability to confer cell survival. Two routinely utilized bone graft materials, anorganic bone matrix (ABM) and ABM carrying the cell binding peptide P-15 were compared for cell viability and apoptosis. Anchorage-dependent human foreskin fibroblasts (HFF) or osteogenic MC3T3-E1 cells were seeded on ABM or ABM/P-15. After serum withdrawal, viability and level of apoptosis were significantly (p<0.05) improved for cells on ABM/P-15 compared to cells on ABM. Scanning electron microscopy confirmed large, spread cells on ABM/P-15 and smaller cells with apoptotic cellular blebs on ABM. In addition, viable cell attachment was significantly greater on cells cultured on ABM/P-15 compared with demineralized freeze-dried bone allograft. This is the first report that the P-15 cell binding peptide, when complexed with ABM, promotes cell survival. The P-15 peptide may modulate cell number and tissue structure by the enhancement of viable cell attachment and via the regulation of apoptosis. PMID- 15120531 TI - Cyclo-(DfKRG) peptide grafting onto Ti-6Al-4V: physical characterization and interest towards human osteoprogenitor cells adhesion. AB - In the present paper, specific interest has been devoted to the design of new hybrid materials associating Ti-6Al-4V alloy and osteoprogenitor cells through the grafting of two RGD containing peptides displaying a different conformation (linear RGD and cyclo-DfKRG) onto titanium surface. Biomimetic modification was performed by means of a three-step reaction procedure: silanization with APTES, cross-linking with SMP and finally immobilization of peptides thanks to thiol bonding. The whole process was performed in anhydrous conditions to ensure homogeneous biomolecules layout as well as to guarantee a sufficient amount of biomolecules grafted onto surfaces. The efficiency of this new route for biomimetic modification of titanium surface was demonstrated by measuring the adhesion between 1 and 24 h of osteoprogenitor cells isolated from HBMSC. Benefits of the as-proposed method were related to the high concentration of peptides grafted onto the surface (around 20 pmol/mm(2)) as well as to the capacity of cyclo-DfKRG peptide to interact with integrin receptors. Moreover, High Resolution beta-imager (using [(35)S]-Cys) has exhibited the stability of peptides grafted onto the surface when treated in harsh conditions. PMID- 15120532 TI - The effect of sample preparation technique on determination of structure and nanomechanical properties of human cementum hard tissue. AB - The mechanical properties of a tissue can be evaluated by determining the response of the structure to mechanical loading. This can be accomplished only when the tissue has been prepared with minimum to no artifacts, thus preserving its structure. In this study it was hypothesized that the structure of cementum is inhomogeneous, contributing to a significant variation in mechanical properties of cementum. Therefore, the goals of the study were to identify potential artifacts generated by conventional sample preparation techniques such as polishing and ultrasectioning and subsequently characterize the prepared specimens using an atomic force microscope (AFM) and an AFM-nanoindenter. Comparisons between cryofractured, ultrasectioned and polished specimens concluded that ultrasectioned surfaces have significantly lower average surface roughness 'R(a)' (p<0.05). Microstructure of ultrasectioned specimens characterized using an AFM illustrated Sharpey's fibers (SF) and intrinsic fibers (IF) running perpendicular and parallel to the root surface similar to the observed microstructure of cryofractured cementum. In addition, a 10-50 microm wide cementum dentin junction (CDJ) was distinctly observed in the ultrasectioned specimens but not in polished specimens. The SF and CDJ illustrated relatively higher levels of hydrophilicity under wet conditions. The observed inhomogeneous microstructure of the ultrasectioned specimens led to a broader range of nanomechanical properties (modulus: 14.2-25.9 GPa; hardness: 0.48-1.09 GPa). However, masking of the same regions such as SF and CDJ due to smeared cementum in polished specimens resulted in a narrower range of nanomechanical properties (modulus: 18.2-20.8 GPa; hardness: 0.79-0.89 GPa). This effect is most noticeable under wet conditions for ultrasectioned specimens (modulus 2.6-10.9 GPa; hardness 0.05-0.30 GPa) compared to the polished specimens (modulus 12.2-14.5 GPa; hardness 0.33-0.45 GPa). Cementum also was shown to be highly viscoelastic, especially when hydrated. The results suggest ultrasectioning of cementum was superior to polishing preparation technique since it allowed visualization of cementum structures similar to cryofractured specimens while providing a flat surface necessary for AFM-based nanoindentation techniques. Additionally, the structural inhomogeneity observed within ultrasectioned cementum contributed to a broader range of mechanical properties. PMID- 15120533 TI - Affective aprosodia from a medial frontal stroke. AB - BACKGROUND AND OBJECTIVES: Whereas injury to the left hemisphere induces aphasia, injury to the right hemisphere's perisylvian region induces an impairment of emotional speech prosody (affective aprosodia). Left-sided medial frontal lesions are associated with reduced verbal fluency with relatively intact comprehension and repetition (transcortical motor aphasia), but persistent affective prosodic defects associated with right medial frontal lesions have not been described. METHODS: We assessed the prosody of a man who sustained a right medial frontal cerebral infarction seven years prior. RESULTS: While propositional speech expression was normal including syntactic prosody, the patient was impaired at expressing emotions using prosody. His comprehension and repetition of prosody were also impaired but less so than expression. CONCLUSIONS: Right medial frontal lesions can induce an affective aprosodia that primarily impairs expression. PMID- 15120534 TI - Unitary vs multiple semantics: PET studies of word and picture processing. AB - In this paper we examine a central issue in cognitive neuroscience: are there separate conceptual representations associated with different input modalities (e.g., Paivio, 1971, 1986; Warrington & Shallice, 1984) or do inputs from different modalities converge on to the same set of representations (e.g., Caramazza, Hillis, Rapp, & Romani, 1990; Lambon Ralph, Graham, Patterson, & Hodges, 1999; Rapp, Hillis, & Caramazza, 1993)? We present an analysis of four PET studies (three semantic categorisation tasks and one lexical decision task), two of which employ words as stimuli and two of which employ pictures. Using conjunction analyses, we found robust semantic activation, common to both input modalities in anterior and medial aspects of the left fusiform gyrus, left parahippocampal and perirhinal cortices, and left inferior frontal gyrus (BA 47). There were modality-specific activations in both temporal poles (words) and occipitotemporal cortices (pictures). We propose that the temporal poles are involved in processing both words and pictures, but their engagement might be primarily determined by the level of specificity at which an object is processed. Activation in posterior temporal regions associated with picture processing most likely reflects intermediate, pre-semantic stages of visual processing. Our data are most consistent with a hierarchically structured, unitary system of semantic representations for both verbal and visual modalities, subserved by anterior regions of the inferior temporal cortex. PMID- 15120535 TI - Can small lesions induce language reorganization as large lesions do? AB - Shift of the cortical mechanisms of language from the usually dominant left to the non-dominant right hemisphere has been demonstrated in the presence of large brain lesions. Here, we report a similar phenomenon in a patient with a cavernoma over the anterolateral superior temporal gyrus associated with epilepsy. Language mapping was performed by two complementary procedures, magnetoencephalography, and electrocorticography. The maps, indicated right temporal lobe dominance for receptive language and left frontal lobe dominance for expressive language. These results indicate that a small lesion, associated with epilepsy, may produce selective shifting of receptive language mechanisms as large lesions have been known to produce. PMID- 15120536 TI - Neural responses to morphological, syntactic, and semantic properties of single words: an fMRI study. AB - Dissociations in the recognition of specific classes of words have been documented in brain-injured populations. These include deficits in the recognition and production of morphologically complex words as well as impairments specific to particular syntactic classes such as verbs. However, functional imaging evidence for distinctions among the neural systems underlying these dissociations has been inconclusive. We explored the neural systems involved in processing different word classes in a functional Magnetic Resonance Imaging study, contrasting four groups of words co-varying morphological complexity (simple, monomorphemic words vs complex derived or inflected words) and syntactic class (verbs vs nouns/adjectives). Subtraction of word from letter string processing showed activation in left frontal and temporal lobe regions consistent with prior studies of visual word processing. No differences were observed for morphologically complex and simple words, despite adequate power to detect stimulus specific effects. A region of posterior left middle temporal gyrus showed significantly increased activation for verbs. Post hoc analyses showed that this elevated activation could also be related to semantic properties of the stimulus items (verbs have stronger action associations than nouns, and action association is correlated with activation). Results suggest that semantic as well as syntactic factors should be considered when assessing the neural systems involved in single word comprehension. PMID- 15120537 TI - Inference revision processing in adults with and without aphasia. AB - Processing abilities in aphasia, and the nature of processing breakdowns, were the focuses of this investigation. Individuals with either fluent or nonfluent aphasia, plus a control group, participated in a cross-modal lexical priming task designed to elicit priming effects when activation of inference interpretations occurred. Comprehension of inferences was measured by responses to four types of questions that related to the inferences. Results indicated that both the control group, as well as the nonfluent aphasia group, activated the intended meaning of the stimuli whereas the fluent aphasics did not. Comprehension of the inferences was best demonstrated by control participants, nonfluent aphasic participants, and fluent aphasic participants, in that order. PMID- 15120538 TI - Measurements of auditory-verbal STM span in aphasia: effects of item, task, and lexical impairment. AB - In the first part of this study, we investigated effects of item and task type on span performance in a group of aphasic individuals with word processing and STM deficits. Group analyses revealed significant effects of item on span performance with span being greater for digits than for words. We also investigated associations between subjects' lexical-semantic and phonological processing abilities and performance on four measures of verbal span (digit and word span, each varied for type of response, verbal vs. pointing) as well as one measure of nonverbal span. We predicted and found that the patterns of association between verbal span tasks and lexical abilities reflected the integrity of language processes and representations deployed in each paradigm used to assess span. Performance on the pointing span task, which engages both lexical-semantic and phonological processes, correlated with measures of both lexical-semantic and phonological abilities. Performance on repetition span, which engages primarily input and output phonological processes, correlated with measures of phonological abilities but not measures of lexical-semantic abilities. However, when partial correlations were performed for two subject groups based on their relative preservation of lexical-semantic ability (less or more than phonological ability), repetition span correlated with lexical-semantic measures only in the subgroup with relatively impaired lexical-semantics. Additionally, performance on the nonverbal span task correlated with measures of phonological abilities, suggesting either a general cognitive deficit affecting verbal and nonverbal STM or possibly, the use of a verbal strategy to perform this task. Our discussion focuses on the interpretation of span measurements in clinical practice and research, as well as the implications of these data for theories of short-term memory and word processing. PMID- 15120539 TI - Semantic, lexical, and phonological influences on the production of verb inflections in agrammatic aphasia. AB - Verb inflection errors, often seen in agrammatic aphasic speech, have been attributed to either impaired encoding of diacritical features that specify tense and aspect, or to impaired affixation during phonological encoding. In this study we examined the effect of semantic markedness, word form frequency and affix frequency, as well as accuracy and error patterns, in an attempt to evaluate whether diacritical or affixation operations are impaired. Verb inflections (V + ing, V + ed, V + s, and V stem in present progressive, past, present 3rd person singular, and future tense contexts, respectively) were elicited in eight mild moderate agrammatic aphasic individuals in a sentence context using a picture description task. Results revealed that the majority of verbs produced were affixed (75%) although accuracy was low (36%). Word form frequency was found to be a significant predictor of the accuracy with which verb inflections were produced; while affix frequency and semantic markedness were not found to influence accuracy. These results suggest that a diacritical deficit is more likely to undermine the production of verb inflections than a affixation deficit, and indicate that when diacritical processes are compromised, word form frequency is likely to influence production of verb inflections in agrammatic aphasia. PMID- 15120540 TI - Degree of illiteracy and phonological and metaphonological skills in unschooled adults. AB - Phonological and metaphonological skills are explored in 97 Brazilian illiterate and semiliterate adults. A simple letter- and word-reading task was used to define the degree of illiteracy. Phonemic awareness was strongly dependent on the level of letter and word reading ability. Phonological memory was very low in illiterates and unrelated to letter knowledge. Rhyme identification was relatively preserved in illiterates and semiliterates, and unrelated to letter and word reading level. Phonetic discrimination (minimal pairs) was fairly good and marginally related to reading ability. These results suggest that phonological sensitivity, phonological memory, rhyme identification, and phonemic awareness are distinctive cognitive processes, and that only phonemic awareness is clearly and strongly dependent on the alphabetical acquisition. PMID- 15120541 TI - New phenomenon of abnormal auditory perception associated with emotional and head trauma: pathological confirmation by SPECT scan. AB - We report the case of an immigrant who suffered from death threats and head trauma while a prisoner of war in Kuwait. Two months later, he began to hear conversations that had taken place previously. These perceptions occurred spontaneously or were induced by the patient's effortful concentration. The single photon emission computerized tomography (SPECT) scan performed during this abnormal perceptual experience indicated an increase in regional cerebral blood flow in the left temporal lobe and in the brainstem. PMID- 15120542 TI - Referential cohesion and logical coherence of narration after closed head injury. AB - A group with closed head injury was compared to neurologically intact controls regarding the referential cohesion and logical coherence of narrative production. A sample of six stories was obtained with tasks of cartoon-elicited story-telling and auditory-oral retelling. We found deficits in the clinical group with respect to referential cohesion, logical coherence, and accuracy of narration. The occurrence of deficits depended on the condition of narrative production and, to some extent, on the particular story used. The primary implications of this study pertain to the attention given by researchers to the feature of discourse production being studied and processing demands of the task. PMID- 15120544 TI - A high-field functional MRI study of quadri-lingual subjects. AB - We assessed six multilingual subjects by functional MRI using a Noun Verb Generation task in four different languages. We hypothesised that the degree of proficiency in each language would be related to the extent of functional activity measured in a region of interest analysis. Proficiency in each language was quantified using two neuropsychological tests. All four languages activated overlapping brain areas, corresponding to the major language regions. The number of activated voxels correlated with proficiency, so that the activated volume increased for languages in which a subject had poorer proficiency. Activation did not appear to be dependent on the age at which the language was learnt. PMID- 15120543 TI - Variability in subcortical aphasia is due to variable sites of cortical hypoperfusion. AB - A variety of fluent and nonfluent aphasias have been reported after left basal ganglia stroke. It has been speculated that this heterogeneity may reflect variations in cortical hypoperfusion resulting from large vessel stenosis. To test this hypothesis, a consecutive series of 24 patients with left caudate infarct identified with diffusion-weighted imaging underwent language testing and perfusion-weighted imaging < 24h from onset of symptoms. Specific regions in perisylvian cortex were rated for the percentage of the region that was hypoperfused. Aphasia type was determined on the basis of speech fluency, comprehension, and repetition performance on the language tests. Association between aphasia type/language impairment and regions of hypoperfusion were identified with Fisher's exact tests. Results demonstrated that in patients with acute left caudate infarct, the presence and type of aphasia reflected regions of hypoperfusion, and generally followed predictions based on chronic lesion studies, regarding anatomical lesions associated with classic aphasia types. PMID- 15120545 TI - Auditory repetition priming is impaired in pure alexic patients. AB - Alexia without agraphia, or "pure" alexia, is an acquired impairment in reading that leaves writing skills intact. Repetition priming for visually presented words is diminished in pure alexia. However, it is not possible to verify whether this priming deficit is modality-specific or modality independent because reading abilities are compromised. Hence, auditory repetition priming was assessed with lexical decision and word stem completion tasks in pure alexic patients with lesions in left inferior temporal-occipital cortex and the splenium. Perceptually based, modality-specific priming models predict intact auditory priming, since auditory association cortex is spared in the patients. Alternatively, modality independent models, which suggest that priming reflects the temporary modification of an amodal system, might predict impairments. Baseline performance was matched in the patients and controls, although lexical decision priming measures showed an interaction between group and repetition lag. The patients showed intact immediate priming but significantly less priming than controls at longer delays. Furthermore, word stem completion priming was abolished in the patients. One explanation for the deficit is that left inferior temporal occipital cortex supports amodal aspects of priming, as suggested by recent neuroimaging results. Another possibility is that long-term auditory priming relies on covert orthographic representations which were unavailable in the patients. The results provide support for interactive models of word identification. PMID- 15120546 TI - Hemispheric specialization and independence for word recognition: a comparison of three computational models. AB - Two findings serve as the hallmark for hemispheric specialization during lateralized lexical decision. First is an overall word advantage, with words being recognized more quickly and accurately than non-words (the effect being stronger in response latency). Second, a right visual field advantage is observed for words, with little or no hemispheric differences in the ability to identify non-words. Several theories have been proposed to account for this difference in word and non-word recognition, some by suggesting dual routes of lexical access and others by incorporating separate, and potentially independent, word and non word detection mechanisms. We compare three previously proposed cognitive theories of hemispheric interactions (callosal relay, direct access, and cooperative hemispheres) through neural network modeling, with each network incorporating different means of interhemispheric communication. When parameters were varied to simulate left hemisphere specialization for lexical decision, only the cooperative hemispheres model showed both a consistent left hemisphere advantage for word recognition but not non-word recognition, as well as an overall word advantage. These results support the theory that neural representations of words are more strongly established in the left hemisphere through prior learning, despite open communication between the hemispheres during both learning and recall. PMID- 15120547 TI - Is word perception in a second language more vulnerable than in one's native language? Evidence from brain potentials in a dual task setting. AB - We assessed the effect of additional tasks on language perception in second language and native speakers. The N400 component of the event-related potential was recorded to spoken nouns that had to be judged for synonymity with a preceding word, while additional choice responses were required to visual stimuli. In both participant groups N400 was delayed as a function of temporal overlap with the additional task. In second-language speakers there was a global delay of N400 independent of additional task load and a decrease of accuracy at highest overlap. These findings are interpreted within a single channel account of language perception. PMID- 15120548 TI - Action verbal fluency normative data for the elderly. AB - An emerging body of literature points to the prominent role of the frontal lobes in the retrieval of verbs, whereas production of common and proper nouns arguably is mediated primarily by posterior and anterior temporal regions, respectively. Although the majority of studies examining the neuroanatomic distinctions between verb and noun retrieval have relied on action naming tasks (naming depicted activities, e.g., running) as indicators of verb retrieval abilities, recent studies have utilized an action (verb) verbal fluency measure, the Action Fluency Test (AFT), to assess verb retrieval. Findings from these studies suggest that action fluency is sensitive to the integrity of fronto-subcortical neural circuitry and that it is a valid measure of executive and language functions. The AFT is an easily administered executive function measure, but no normative data for the AFT or other action fluency tasks has been published. This study was undertaken to provide initial AFT normative data for a sample of 145 healthy elderly subjects. As education is significantly correlated with AFT scores, the normative data are stratified by educational level. PMID- 15120550 TI - Perception of words and non-words in the upper and lower visual fields. AB - The findings of previous investigations into word perception in the upper and the lower visual field (VF) are variable and may have incurred non-perceptual biases caused by the asymmetric distribution of information within a word, an advantage for saccadic eye-movements to targets in the upper VF and the possibility that stimuli were not projected to the correct retinal locations. The present study used the Reicher-Wheeler task and an eye-tracker to show that, using stringent methodology, a right over left VF advantage is observed for word recognition, but that no differences were found between the upper and the lower VF for either word or non-word recognition. The results are discussed in terms of the neuroanatomy and perceptual abilities of the upper and the lower VF and implications for other studies of letter-string perception in the upper and the lower VF are presented. PMID- 15120549 TI - Gapping: Electrophysiological evidence for immediate processing of "missing" verbs in sentence comprehension. AB - In the present study we use event related potentials (ERPs) to explore the time course of identification and resolution of verb gaps. ERPs were recorded while participants read sentences that contained a verb gap like Ron took/sanded the planks, and Bill O the hammer... Plausibility of the critical words (hammer) that followed the verb gap was manipulated. Relative to the plausible control (preceded by took), ERPs to the critical word in the implausible condition (preceded by sanded) showed an N400, followed by a positivity (P600). ERPs to determiners following gapped verbs showed a negativity between 100 and 300 ms, and a positivity between 300 and 500 ms compared to determiners in non-gapping constructions. These results suggest that the sentence processor recognizes a verb gap and reconstructs the verb information at the earliest possible occasion, and that this reconstruction process is different from the reconstruction of antecedents in other filler-gap constructions (e.g., WH gaps). PMID- 15120551 TI - Symbol-string sensitivity and children's reading. AB - In this study of primary school children, a novel 'symbol-string' task is used to assess sensitivity to the position of briefly presented non-alphabetic but letter like symbols. The results demonstrate that sensitivity in the symbol-string task explains a unique proportion of the variability in children's contextual reading accuracy. Moreover, developmental dyslexic readers show reduced sensitivity in this task, compared to chronological age controls. The results suggest that limitations set by visuo-spatial processes and/or attentional iconic memory resources may constrain children's reading accuracy. PMID- 15120552 TI - Regularity re-revisited: modality matters. AB - The issue of regular-irregular past tense formation was examined in a cross-modal lexical decision task in Modern Greek, a language where the orthographic and phonological overlap between present and past tense stems is the same for both regular and irregular verbs. The experiment described here is a follow-up study of previous visual lexical decision experiments (Tsapkini, Kehayia, & Harema, 2002) that also addressed the regular-irregular distinction in Greek. In the present experiment, we investigated the effect of input modality in lexical processing and compared different types of regular and irregular verbs. In contrast to our previous intra-modal (visual-visual) priming experiments, in this cross-modal (auditory-visual) priming study, we found that regular verbs with an orthographically salient morphemic aspectual marker elicited the same facilitation as those without an orthographically salient marker. However, irregular verbs did not exhibit a different priming pattern with respect to modality. We interpret these results in the framework of a two-level lexical processing approach with modality-specific access representations at a surface level and modality-independent morphemic representations at a deeper level. PMID- 15120553 TI - Learning new names for new objects: cortical effects as measured by magnetoencephalography. AB - We tracked the evolvement of naming-related cortical dynamics with magnetoencephalography when five normal adults successfully learned names and/or meanings of unfamiliar objects. In all subjects, the learning of new names was associated with pronounced cortical effects. The learning effect was of long latency and emerged as a change of activation in the same cortical network that was active during naming of familiar items. In four out of five subjects, the cortical learning effect occurred in the inferior parietal lobe. In three of these subjects, the cortical effect was left-sided. These results suggest that the inferior parietal lobe plays an important role in the acquisition of novel words, presumably as a part of working memory systems. PMID- 15120554 TI - Selective removal of the heavy metal ions from waters and industrial wastewaters by ion-exchange method. AB - By ion exchange undesirable ions are replaced by others which don't contribute to contamination of the environment. The method is technologically simple and enables efficient removal of even traces of impurities from solutions. Examples of selective removal of heavy metal ions by ion-exchange are presented. They include removal of Pb(II), Hg(II), Cd(II), Ni(II), V(IV,V), Cr(III,VI), Cu(II) and Zn(II) from water and industrial wastewaters by means various modern types of ion exchangers. PMID- 15120555 TI - Petroleum hydrocarbons and polycyclic aromatic hydrocarbons in the surficial sediments of Xiamen Harbour and Yuan Dan Lake, China. AB - Surficial sediments were sampled from nine stations in Xiamen Harbour and two stations in Yuan Dan Lake during April 2002. Sediment samples were extracted by organic solvents, separated by silica gel column chromatography and analyzed by gas chromatography-mass selective detector (GC-MSD). Selected ion monitoring was at M/Z=57 for petroleum hydrocarbons (PHCs) and individual M/Zs for each of the 15 typical polycyclic aromatic hydrocarbons (PAHs) and nine alkylated PAHs. The results showed that concentrations of PHCs and total PAHs in the sediments of Yuan Dan Lake were 1397 microg g(-1) (dry weight, dw) and 1377 ng g(-1) (dw), respectively. The ranges for PHCs and total PAHs in the sediments from Xiamen Harbour were 133-943 microg g(-1) (dw) and 98-309 ng g(-1) (dw), respectively. Shipping activities, industrial wastewater discharges, fuel oil spillage from ships and vehicles were the main sources of PHCs and PAHs in the Harbour. In addition, the widespread use of coal for industrial processes and domestic consumption accounted for the second largest source of PAHs in the sediments, while atmospheric transport and deposition of PAHs are also important. PMID- 15120556 TI - Fate of biodegradable dissolved organic carbon produced by ozonation on biological activated carbon. AB - The adsorption and desorption characteristics of BDOC produced by ozonation and the replacement of BDOC by non-BDOC on BAC was studied. The fate of BDOC produced by ozonation in the BAC column was also evaluated by comparative experiment between the BAC supplied with the mixture of BDOC and non-BDOC and the BAC supplied only with non-BDOC. Fulvic acids extracted from two river sediments and one forest soil were used. BDOC produced by ozonation showed the same or lower adsorption capacity than non-BDOC after biodegradation. The adsorption rate of BDOC on GAC was lower than that of non-BDOC. BDOC produced by ozonation had low desorbability and majority of BDOC produced were not replaced by non-BDOC. BDOC in the ozonated fulvic acid did not affect the breakthrough of the ozonated fulvic acid on GAC in the early stage of adsorption, suggesting that most of BDOC were biodegraded on the surface of GAC before adsorption. Therefore, the production of BDOC by ozonation before the GAC treatment is very effective for the extension of GAC service life and the reduction of DOC loading to GAC. PMID- 15120557 TI - Metabolism of cyanide by Chinese vegetation. AB - Cyanide is a high-volume production chemical and the most commonly used leaching reagent for gold and silver extraction. Its environmental behavior and fate is of significant concern because it is a highly toxic compound. Vascular plants possess an enzyme system that detoxifies cyanide by converting it to the amino acid asparagine. This paper presents an investigation of the potential of Chinese vegetation to degrade cyanide. Detached leaves (1.5 g fresh weight) from 28 species of 23 families were kept in glass vessel with 100 ml of aqueous solution spiked with potassium cyanide at 23.5 degrees C for 28 h. Cyanide concentrations ranged from 0.83 to 1.0 CN mg l(-1). The disappearance of cyanide from the aqueous solution was analyzed spectrophotometrically. The fastest cyanide removal was by Chinese elder, Sambucus chinensis, with a removal capacity of 8.8 mg CN kg(-1) h(-1), followed by upright hedge-parsley (Torilis japonica) with a value of 7.5 mg CN kg(-1) h(-1). The lowest removal capacity had the snow-pine tree (Credrus deodara (Roxb.) Loud). Results from this investigation indicated that a wide range of plant species is able to efficiently metabolize cyanide. Therefore, cyanide elimination with plants seems to be a feasible option for cleaning soils and water contaminated by cyanide from gold and silver mines or from other sources. PMID- 15120558 TI - Effect of additional materials on the properties of glass-ceramic produced from incinerator fly ashes. AB - There are 21 Metro-waste incinerators in Taiwan under construction and are expected to be finished at year 2003. It is estimated that these incinerators will produce about two million tons of incinerator ash. In order to reduce the volume and eliminate contamination problems, high temperature molten technology studies have been conducted. The purpose of this research was that of trying to control the chemical composition of the glass-ceramic produced from incinerator fly ash, in order to improve the characteristics of the glass-ceramic. The experimental results showed that the additional materials, Mg(OH)2 and waste glass cullet, can change glass-ceramic phases from gehlenite to augite, pigeonite, and diopside. The physical, mechanical and chemical resistance properties of the glass-ceramic also showed much better characteristics than prepared glass-ceramic using incinerator fly ash alone. PMID- 15120559 TI - Decomposition of 2-mercaptothiazoline in aqueous solution by ozonation. AB - This study investigates the ozonation of 2-mercaptothiazoline (2-MT). The 2-MT is one of the important organic additives for the electroplating solution of the printed wiring board industry and has been widely used as a corrosion inhibitor in many industrial processes. It is of concern for the aquatic pollution control especially in the wastewaters. Semibatch ozonation experiments in the completely stirred tank reactor are performed under various concentrations of input ozone. The concentrations of 2-MT, sulfate, and ammonium are analyzed at specified time intervals to elucidate the decomposition of 2-MT during the ozonation. In addition, the time variation of the dissolved ozone concentration (C(ALb)) is continuously monitored in the course of experiments. Total organic carbon (TOC) is chosen and measured as a mineralization index of the ozonation of 2-MT. The results indicate that the decomposition of 2-MT is efficient, while the mineralization of TOC is limited via the ozonation only. Simultaneously, the yield of sulfate with the maximum value of about 47% is characterized by the increases of TOC removal and ozone consumption. These results can provide some useful information for assessing the feasibility of the treatment of 2-MT in the aqueous solution by the ozonation. PMID- 15120560 TI - Competitive adsorption characteristics of Co2+, Ni2+, and Cr3+ by IRN-77 cation exchange resin in synthesized wastewater. AB - Adsorption properties of Co2+, Ni2+, and Cr3+ on an Amberlite IRN-77 cation exchange resin were investigated in batch systems. Levels of adsorption rapidly approached an equilibrium state within 1 h. The adsorption characteristics of each metal onto the resin were accurately represented by Langmuir isotherms. Co2+ and Ni2+, which have an equivalent electrovalence, displayed similar levels of adsorption onto the resin when they coexisted in the solution. However, when Cr3+ was added to the solution it competitively replaced Co2+ and Ni2+ ions that had been previously adsorbed onto the resin, resulting in the desorption of these metals into the solution. The result was likely due to a higher adsorption affinity of Cr3+ relative to Co2+ and Ni2+. This implies that interactively competitive adsorption of multi-cations onto the resin should be thoroughly considered when contemplating the efficient operation of an ion exchange process in the treatment of industrial wastewater. PMID- 15120561 TI - The enhancement of the biodegradability of phenolic solution using preozonation based on high ozone utilization. AB - In this research, the effects of preozonation on the biodegradability of 4 cresol, 4-nitrophenol and 2-chlorophenol solutions were investigated using a new gas-inducing reactor with high ozone utilization rate. The extent of preozonation may be monitored by determining the characteristic ozonation behaviors of preozonized phenolic solutions, such as residual phenolic concentration, ADMI value and ozone gas outlet concentration. Experimental results showed that as the initial phenolic compounds decomposed completely, the ozone gas outlet concentration rapidly increases. In addition, at pH 7, a peak ADMI value appears during the preozonation of 4-cresol and 2-chlorophenol, while for 4-nitrophenol the ADMI value decreases monotonically. Based on the characteristic ozonation behaviors and the ozone utilization rate, three characteristic times were chosen in order to have better control on the extent of preozonation. The effect of preozonation on the biodegradability of preozonized phenolic solution was studied based on these characteristic times. The intermediate products during the preozonation were also identified. The variation of BOD5 is strongly dependent on the accumulation of intermediate products. It is suggested that the best characteristic time is as the rapid increase of ozone gas outlet concentration in this study. The biodegradability (BOD5/COD) of preozonized 4-cresol, 2 chlorophenol and 4-nitrophenol solutions increase to 0.18, 0.26 and 0.33, respectively, for the best characteristic time. PMID- 15120562 TI - Enhanced degradation of fluorene in soil slurry by Absidia cylindrospora and maltosyl-cyclodextrin. AB - This study investigates the fungal biodegradation of fluorene, a polycyclic aromatic hydrocarbon, in liquid medium and soil slurry. Fungal strains and cyclodextrins were used in order to degrade fluorene and optimize fluorene bioavailability and degradation in soil slurries. After a procedure of selection in solid and liquid media, maltosyl-cyclodextrin, a branched cyclodextrin was chosen. 47 fungal strains isolated from a contaminated site were tested for biodegradation. Results showed the greater efficiency of "adapted" fungi isolated from contaminated soil vs reference strains belonging to the collection of the laboratory. These assays allowed us to select the most efficient strain, Absidia cylindrospora, which was used in a bioaugmentation process. Bioaugmentation tests were performed in an artificially contaminated non-sterile soil. In the presence of A. cylindrospora, more than 90% of the fluorene was degraded within 288 h, while 576 h were necessary in the absence of fungal bioremediation. It also appeared that biodegradation was enhanced by amendment with previously selected maltosyl-cyclodextrin. The results of this study indicate that A. cylindrospora and maltosyl-cyclodextrin could be used successfully in fluorene bioremediation systems. PMID- 15120563 TI - Two stage biological treatment of a diazo reactive textile dye and the fate of the dye metabolites. AB - A two stage anaerobic/aerobic bacterial process was used to decolorize and partially mineralize a reactive vinyl sulfone diazo dye C.I. Reactive Black 5 (RB5) in a synthetic wastewater. Since the anchor group of reactive dyes reacts during the dyeing process, the effect the degree of hydrolysis of the vinyl sulfone dye had on decolorization, mineralization and toxicity in each stage was investigated. An overall color removal of approximately 65% was found for both the fully and partially hydrolyzed dye. Partial mineralization of the fully hydrolyzed RB5 was achieved in the two stage rotating disc reactors. While the anchor group metabolite p-aminobenzene-2-hydroxyethylsulfonic acid (p-ABHES) was mineralized, an oxidized form of the center metabolite (1,2-ketimino-7-amino-8 hydroxynaphthalene-3,6-disulfonic acid) remained in the aerobic stage effluent, causing the effluent to be colored although no RB5 was present. Partially hydrolyzed dye in the influent with vinyl forms of the anchor group caused cessation of biogas production and a reduction in decolorization efficiency in the anaerobic stage. No evidence for mineralization of the partially hydrolyzed dye or its metabolites was found. A method for evaluating dye mineralization using lumped parameters is presented. PMID- 15120564 TI - Evaluation of modified clay coagulant for sewage treatment. AB - The use of modified clays as coagulants for sewage treatment was investigated in this study. The raw clays were montmorillonites K10 and KSF, and were modified by polymeric Al or Fe and/or Al/Fe mixing polymeric species. The comparative performance of modified clays and aluminium sulphate and ferric sulphate were evaluated in terms of the removal of turbidity, suspended solids, UV(254)-abs, colour, and total and soluble CODs. The results demonstrated that after being modified with mixing polymeric Al/Fe species, two montmorillonite clays possess greater properties to remove the particles (as suspended solids) and organic pollutants (as COD and UV(254)-abs) from the sewage and to enhance the particle settling rate significantly. PMID- 15120565 TI - Electrochemical reduction of organohalogen compound by noble metal sintered electrode. AB - 1,2,3-Trichlorobenzene (1,2,3-TCB) was used as a model sample of persistent organic pollutants (POPs) which was dechlorinated by a closed electrochemical reduction system under an inert gas atmosphere. The effect of the electrode material was examined in the reaction. Dechlorination yields in different type of cathode electrodes using sintered RuO2 (major)/Pt/PdO, sintered Pt(major)/IrO2/RuO2, sintered RuO2, sintered PdO, sintered Pt, sintered PdO/Pt, sintered Pd/Pt and plain Pd plate were 91%, 81%, 59%, 96%, 53%, 97%, 82% and 70% respectively, at reaction times of 60 or 120 min. The reaction was exothermic after initially starting at room temperature. This electrochemical reduction system was friendly technology for environment using cation exchange membrane, supplying sodium ions from sodium hydroxide solution as anolyte. Trace amounts of dichlorobenzene, as products of stepwise dechlorination, were observed with different pathways, depending on the electrode material. Electrodes with Ru and Pd were selective mainly for meta-position dechlorination, while those with Pt groups selective mainly for ortho-position (o-position) dechlorination. A PdO sintered electrode had an especially high selectivity for meta-position (m position) dechlorination. The results suggest that dechlorination is an electrocatalytic reduction in this cation supply system. PMID- 15120566 TI - COX-2 and iNOS in opioid-induced delayed cardioprotection in the intact rat. AB - Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) have been previously implicated in the late phase of cardioprotection associated with opioid-induced and ischemic preconditioning (IPC) in conscious rabbits and COX-2 in isolated rat hearts pretreated with an exogenous delta opioid agonist. However, it is not know if both iNOS and COX-2 mediate the late phase of cardioprotection induced by opioids in the intact blood-perfused rat. Therefore, we investigated the role of COX-2 and iNOS in the delayed phase of protection mediated by delta opioid receptor activation. Rats were pretreated 24 hours prior to an occlusion/reperfusion protocol with the selective non-peptide delta opioid agonists, BW373U86 (BW) and SNC-121 (SNC). NS-398, a selective COX-2 inhibitor was administered after the 24-hour recovery period just prior to index ischemia. The selective iNOS inhibitors, S-methylthiourea (SMT) and aminoguanidine (AG), were administered in conjunction with opioid pretreatment or were also given 24 hours after opioid administration just prior to index ischemia. COX-2 inhibition by NS-398 given 24 hours after opioid administration attenuated the protective effects of both BW and SNC (46 +/- 6 vs. 13 +/- 3 and 51 +/- 5 vs. 29 +/- 2, p < 0.001, respectively). Similarly, inhibition of iNOS following 24 hours of treatment with opioids also attenuated the protective effects of BW and SNC. However, the delayed protective effects of the opioids were not attenuated by pretreatment with the iNOS inhibitors 24 hours prior to the infarct protocol. These results suggest that both COX-2 and iNOS are mediators of delayed protection induced by non-peptide delta opioid agonists. It appears that the trigger effect is not dependent on the activity of iNOS or COX-2 but the late phase of cardioprotection is dependent on the upregulation of these enzymes. PMID- 15120567 TI - Effects of modulators of N-methyl-D-aspartate receptor-mediated neurotransmission on diazepam discrimination in rats. AB - N-methyl-D-aspartate (NMDA) antagonists share a number of pharmacological effects with GABA(A) agonists, including anxiolytic and anticonvulsant effects. This study evaluated the effects of site-selective NMDA antagonists in rats trained to discriminate the benzodiazepine diazepam from vehicle. As expected, diazepam produced robust discriminative stimulus effects and dose-dependently substituted for the training dose. Mixed results were obtained with competitive NMDA antagonists: whereas NPC 17742 partially substituted for diazepam, SDZ EAA 494 did not elicit responding on the diazepam-associated lever. Other site-selective NMDA antagonists, including the open channel blocker phencyclidine, the glycine site antagonists ACEA 1021 and MDL 102,288, the polyamine-site antagonist arcaine, and the glutamate release inhibitor riluzole, failed to substitute for diazepam. Agonists at nonbenzodiazepine sites of the GABA(A) receptor complex were also tested for comparison purposes. The barbiturate pentobarbital and the neurosteroid Co 2-1068 partially substituted for diazepam. In contrast, the anticonvulsant carbamazepine failed to substitute even at a dose that substantially reduced response rates. These results suggest that substitution of NMDA antagonists for GABA(A) agonists is dependent upon the site at which the NMDA antagonist binds. Further, they suggest that similarities between the stimulus properties of GABA(A) agonists and NMDA antagonists are at least as strong as similarities among agonists acting at different sites on GABA(A) receptors. PMID- 15120568 TI - Sensitization of recombinant vanilloid receptor-1 by various neurotrophic factors. AB - The vanilloid receptor (VR1) is a central integrator molecule of nociceptive stimuli. In this study, we have measured the effects of various neurotrophins (nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3, and -4) on recombinant rat VR1-mediated intracellular calcium rise in response to capsaicin in VR1/C6 cells. Our results clearly show that all neurotrophins sensitize the VR1 to capsaicin. Furthermore, using K252a, an inhibitor of tyrosine kinases, we present that actions of neurotrophins are mediated by the trk (A, B, C) receptors expressed in these cells. These data argue for the putative roles of neurotrophins in inducing inflammatory (thermal) hyperalgesia via VR1. PMID- 15120569 TI - Orally administered rosmarinic acid is present as the conjugated and/or methylated forms in plasma, and is degraded and metabolized to conjugated forms of caffeic acid, ferulic acid and m-coumaric acid. AB - Rosmarinic acid (RA) is contained in various Lamiaceae herbs used commonly as culinary herbs. Although RA has various potent physiological actions, little is known on its bioavailability. We therefore investigated the absorption and metabolism of orally administered RA in rats. After being deprived of food for 12 h, RA (50 mg/kg body weight) or deionized water was administered orally to rats. Blood samples were collected from a cannula inserted in the femoral artery before and at designated time intervals after administration of RA. Urine excreted within 0 to 8 h and 8 to 18 h post-administration was also collected. RA and its related metabolites in plasma and urine were measured by LC-MS after treatment with sulfatase and/or beta-glucuronidase. RA, mono-methylated RA (methyl-RA) and m-coumaric acid (COA) were detected in plasma, with peak concentrations being reached at 0.5, 1 and 8 h after RA administration, respectively. RA, methyl-RA, caffeic acid (CAA), ferulic acid (FA) and COA were detected in urine after RA administration. These components in plasma and urine were present predominantly as conjugated forms such as glucuronide or sulfate. The percentage of the original oral dose of RA excreted in the urine within 18 h of administration as free and conjugated forms was 0.44 +/- 0.21% for RA, 1.60 +/- 0.74% for methyl RA, 1.06 +/- 0.35% for CAA, 1.70 +/- 0.45% for FA and 0.67 +/- 0.29% for COA. Approximately 83% of the total amount of these metabolites was excreted in the period 8 to 18 h after RA administration. These results suggest that RA was absorbed and metabolized as conjugated and/or methylated forms, and that the majority of RA absorbed was degraded into conjugated and/or methylated forms of CAA, FA and COA before being excreted gradually in the urine. PMID- 15120570 TI - Benzamide riboside induced mitochondrial mediated apoptosis in human lung cancer H520 cells. AB - Benzamide riboside (BR) is a novel anticancer agent exhibiting pronounced activity against several human tumor cell lines via the inhibition of inosine 5' monophosphate dehydrogenase (IMPDH), thereby restricting the biosynthesis of guanylates. Although it has been demonstrated that BR inhibits IMPDH and induces apoptosis, however, not much attention has been directed to the mechanism of apoptosis induction by this compound. The purpose of the present investigation was to investigate the mechanism of cytotoxicity induced by BR in human lung cancer cells. Non-small cell lung cancer [NSCLC] is the most prevalent type of lung cancer especially in India, and displays resistance to anticancer treatment. The results reveal that BR at a dose of 50 microM induces apoptosis in NSCLC H520 cells. This was ascertained by alteration in cellular morphology, TUNEL assay and flow cytometry. While Bax protein level was unaffected there was down regulation of anti-apoptotic Bcl-2 protein and up regulation of p53 as observed by Western blotting. Induction of apoptosis was accompanied by significant increase in caspase-3 activity. BR is a potent growth inhibitory pro-drug rationally synthesized to mimic NAD and inhibits PARP at high concentrations when assayed in permeabilized leukemic cells. Our observations showed that increased caspase-3 activity was accompanied by PARP cleavage. We also observed release of cytochrome c from mitochondria to the cytosol whereas no change was seen in the levels of apoptosis inducing factor (AIF). These findings indicate that BR induces apoptosis in H520 cells via the intrinsic mitochondrial pathway. PMID- 15120571 TI - Antibacterial activity of ethanolic and aqueous extracts of Acacia aroma Gill. ex Hook et Arn. AB - The purpose of the present study was to investigate the antibacterial activity of seven ethanolic extracts and three aqueous extracts from various parts (leaves, stems and flowers) of A. aroma against 163 strains of antibiotic multi-resistant bacteria. The disc diffusion assay was performed to evaluate antibacterial activity of the A. aroma crude extracts, against several Gram-positive bacteria (E. faecalis, S. aureus, coagulase-negative stahylococci, S. pyogenes, S. agalactiae, S. aureus ATCC 29213, E. faecalis ATCC 29212) and Gram-negative bacteria (E. coli., K. pneumoniae, P. mirabilis, E. cloacae, S. marcescens, M morganii, A. baumannii, P. aeruginosa, S. maltophilia, E. coli ATCC 35218, P. aeruginosa ATCC 27853, E. coli ATCC 25922). All ethanolic extracts showed activity against gram-positive bacteria. Among all obtained extracts, only leaf and flower fluid extracts showed activity against Gram-negative bacteria. Based on this bioassay, leaf fluid extracts tended to be the most potent, followed by flower fluid extracts. Minimal inhibitory concentration (MIC) values of extracts and antibiotics were comparatively determined by agar and broth dilution methods. Both extracts were active against S. aureus, coagulase-negative stahylococci, E. faecalis and E. faecium and all tested Gram-negative bacteria with MIC values from 0.067 to 0.308 mg/ml. In this study the minimal bactericidal concentration (MBC) values were identical or twice as high than the corresponding MIC for leaf extracts and four or eight times higher than MIC values for flower extracts. This may indicate a bactericidal effect. Stored extracts have similar antibacterial activity as recently obtained extracts. The A. aroma extracts of leaves and flowers may be useful as antibacterial agents against Gram- negative and Gram positive antibiotic multi-resistant microorganisms. PMID- 15120572 TI - Latent antithrombin does not affect physiological angiogenesis: an in vivo study on vascularization of grafted ovarian follicles. AB - Latent antithrombin (L-AT), a heat-denatured form of native antithrombin (AT), is a potent inhibitor of pathological tumor angiogenesis. In the present study, we have investigated whether L-AT has comparable antiangiogenic effects on physiological angiogenesis of ovarian tissue. For this purpose, preovulatory follicles of Syrian golden hamsters were mechanically isolated and transplanted into dorsal skinfold chambers chronically implanted in L-AT- or AT-treated hamsters. Non-treated animals served as controls. Over 14 days after transplantation neovascularization of the follicular grafts was assessed in vivo by quantitative analysis of the newly developed microvascular network, its microvessel density, the diameter of the microvessels, their red blood cell velocity and volumetric blood flow as well as leukocyte-endothelial cell interaction using fluorescence microscopic techniques. In each group, all of the grafted follicles were able to induce angiogenesis. At day 3 after transplantation, sinusoidal sacculations and capillary sprouts could be observed, finally developing complete glomerulum-like microvascular networks within 5 to 7 days. Overall revascularization of grafted follicles did not differ between the groups studied. Interestingly, follicular grafts in L-AT- and AT-treated hamsters presented with higher values of microvessel diameters and volumetric blood flow, when compared to non-treated controls, which may be best interpreted as a reactive response to an increased release of vasoactive mediators. In conclusion, the present study demonstrates, that L-AT has no adverse effects on physiological angiogenesis of freely transplanted ovarian follicles. Thus, L-AT may be an effective drug in tumor therapy, which blocks tumor growth by selective suppression of tumor vascularization without affecting new vessel formation in the female reproductive system. PMID- 15120573 TI - Cardioprotection and myocardial salvage by a disodium disuccinate astaxanthin derivative (Cardax). AB - Cardioprotection in humans by carotenoids has been inferred from observational and epidemiologic studies, however, direct studies of cardioprotection and myocardial salvage by carotenoids are lacking. In the current study, intravenous (I.V.) pre-treatment with a novel carotenoid derivative (disodium disuccinate astaxanthin; Cardax) was evaluated as a myocardial salvage agent in a Sprague Dawley rat infarct model. Animals were dosed once per day I.V. by tail vein injection for 4 days at one of 3 doses (25, 50, and 75 mg/kg) prior to the infarct study carried out on day 5. The results were compared with control animals treated with saline vehicle. Thirty (30) minutes of occlusion of the left anterior descending (LAD) coronary artery was followed by 2 hours of reperfusion prior to sacrifice, a regimen which resulted in a mean infarct size (IS) as a percent (%) of the area at risk (AAR) of 59 +/- 3%. Area at risk was quantified by Patent blue dye injection, and infarct size (IS) was determined by triphenyltetrazolium chloride (TTC) staining. Cardax at 50 and 75 mg/kg for 4 days resulted in a significant mean reduction in IS/AAR to 35 +/- 3% (41% salvage) and 26 +/- 2% (56% salvage), respectively. Infarct size and myocardial salvage were significantly, and linearly, correlated with plasma levels of non esterified, free astaxanthin at the end of reperfusion. These results suggest that parenteral Cardax may find utility in those clinical applications where pre treatment of patients at risk for myocardial infarction is performed. PMID- 15120574 TI - [(3)H]Adenosine uptake in brainstem membranes of CD-1 mice lacking the adenosine A(2a) receptor. AB - Previous studies in our laboratory have demonstrated a decrease in [(3)H]nitrobenzylthioinosine binding sites in the brainstem of adenosine A(2a) receptor knockout mice, particularly in the brain nuclei involved in central control of cardiovascular function [Brain Research 877 (2000) 160]. The present study aimed to correlate this decrease, shown using autoradiography, with a functional change using a previously described method of [(3)H]adenosine uptake in a membrane preparation from the brainstem of wildtype CD - 1 and homozygous mutant mice lacking the adenosine A(2a) receptor. A statistically significant decrease was shown in the mean V(MAX) value obtained from homozygous mutant preparations (4.7 +/- 1.3 fmol/mg protein/20 s, P < 0.05, n = 4) compared to that obtained from wildtype controls (51.6 +/- 4.2 fmol/mg protein/20 s, n = 4). Competition studies using nucleoside uptake inhibitors showed a statistically significant increase in the log IC(50) values for dipyridamole (Wildtype: -4.3 +/ 0.2, Homozygous mutant: -8.3 +/- 0.4, n=5, P < 0.05) and dilazep (Wildtype: -3.9 +/- 0.8, Homozygous mutant: -8.3 +/- 0.8, n=5, P < 0.05) in the preparations using homozygous mutant tissue. The present study, in conjunction with the results of previous studies [Brain Research 877 (2000) 160], indicates that components of purinergic neurotransmission system have apparently adjusted in compensation for the lack of the A(2a) receptor. PMID- 15120575 TI - Nano red elemental selenium has no size effect in the induction of seleno-enzymes in both cultured cells and mice. AB - We previous reported that a nano red elemental selenium (Nano-Se) in the range from 20 approximately 60 nm had similar bioavailability to sodium selenite (BioFactors 15 (2001) 27). We recently found that Nano-Se with different size had marked difference in scavenging an array of free radicals in vitro, the smaller the particle, the better scavenging activity (Free Radic. Biol. Med. 35 (2003) 805). In order to examine whether there is a size effect of Nano-Se in the induction of Se-dependent enzymes, a range of Nano-Se (5 approximately 200 nm) have been prepared based on the control of elemental Se atom aggregation. The sizes of Nano-Se particles were inversely correlated with protein levels in the redox system of selenite and glutathione. Different sizes of red elemental Se were prepared by adding varying amount of bovine serum albumin (BSA). Three different sizes of Nano-Se (5 approximately 15 nm, 20 approximately 60 nm, and 80 approximately 200 nm) have been chosen for the comparison of biological activity in terms of the induction of seleno-enzyme activities. Results showed that there was no significant size effect of Nano-Se from 5 to 200 nm in the induction of glutathione peroxidase (GPx), phospholipid hydroperoxide glutathione peroxidase (PHGPx) and thioredoxin reductase-1 (TrxR-1) in human hepatoma HepG2 cells and the livers of mice. PMID- 15120576 TI - Dual role of polymorphonuclear neutrophils on the growth of Ehrlich ascites tumor (EAT) in mice. AB - We show that granulocytes (PMN) have a dual role in the development of Ehrlich Ascites Tumor (EAT) in mice. EAT intraperitoneal inoculation causes a local inflammatory reaction, ascites development and mortality that distinguish resistant and susceptible strains. In resistant mice (CAF1), there is a less pronounced PMN influx after EAT inoculation than in susceptible Swiss mice. Accordingly, the increase in peritoneal PMN numbers enhanced tumor growth in CAF1 mice, but had no effect in the susceptible Swiss animals. Contrastingly, PMN depletion had no effect in resistant mice but facilitated tumor growth in susceptible animals. Though no differences were noted between the strains in peritoneal cell spreading and hydrogen peroxide release after tumor inoculation, in vitro PMN cytotoxic activity against EAT was significantly higher in susceptible Swiss mice. These data indicate a paradoxical dual role for PMN against EAT: while they help control tumor development in susceptible animals, they seem to enhance tumor growth in resistant mice. PMID- 15120577 TI - Increased beta-secretase activity and expression in rats following transient cerebral ischemia. AB - The aberrant proteolytic processing of the amyloid precursor protein (APP) by beta- and gamma-secretases is key to amyloid plaque formation in Alzheimer's disease (AD). Identification of an aspartyl protease as the beta-secretase (beta site APP cleaving enzyme, BACE) involved in APP processing provides a pharmaceutical target for potential AD treatment. In the present studies, we demonstrate that transient cerebral ischemia in female rats caused a 30% increase in beta-secretase activity. alpha-Secretase activity did not increase significantly. We examined protein levels of BACE1, and its analogue BACE2, in ischemic brain extracts. BACE1 protein levels increased 67%, while BACE2 protein level did not change after such a transient ischemia. Immunohistochemical studies demonstrated that BACE1 protein was increased in the ischemic neocortex, when compared with its contralateral cortex. Further, colocalization assessment indicated that BACE1 strongly associated with staining for the apoptotic marker, TUNEL. These results may partially explain epidemiological study, which demonstrate a higher incidence of dementia after stroke. Further, our results support the hypothesis that apoptosis and aberrant APP processing are correlated events in AD brain, and suggest that inhibition of BACE may have a therapeutic effect in the prevention of dementia after stroke recovery. PMID- 15120578 TI - Neuroprotective effect of fraxetin and myricetin against rotenone-induced apoptosis in neuroblastoma cells. AB - Rotenone-induced apoptosis is considered to contribute to the etiology of Parkinson's disease (PD). We try to prevent the apoptosis induced by rotenone toxicity with 50 microM myricetin, 100 microM fraxetin and 100 microM N acetylcysteine (NAC) that protect against reactive oxygen species (ROS), on SH SY5Y human neuroblastoma cell line. Morphological changes induced by rotenone and intracellular ROS were assessed in live SH-SY5Y dopaminergic cells by confocal microscopy using the fluorescent dyes, dihydroethidium and 2',7' dichlorofluorescein diacetate (DCFH-DA). DNA fragmentation was assayed as index of apoptosis. We also investigated oxidative stress parameters such as the glutathione redox status and lipid peroxidation. The exposure of the SH-SY5Y cells to rotenone 5 microM for 16 h produced severe morphological changes, DNA fragmentation and significative increases in the levels of hydrogen peroxide and superoxide anion. These increases were reduced by a 30-min pretreatment with fraxetin 100 microM or NAC 100 microM. DNA laddering produced by rotenone treatment was also inhibited by fraxetin and NAC. Treatment with 5 microM rotenone induced loss of reduced glutathione (GSH) and increased cellular levels of oxidized glutathione (GSSG). Fraxetin and NAC treatments restored glutathione redox ratio diminished after rotenone challenge and decreased the levels of lipid peroxidation. These results suggest that the natural antioxidants, such as fraxetin, may prevent the apoptotic death of dopaminergic cells induced by rotenone and mediated by oxidative stress. PMID- 15120579 TI - Selection for pentobarbital withdrawal severity: correlated differences in withdrawal from other sedative drugs. AB - In mice, withdrawal from agents that depress central nervous system function, such as barbiturates and benzodiazepines, results in the production of a withdrawal syndrome, one feature of which is increased severity of handling induced convulsions (HICs). High and Low Pentobarbital Withdrawal mice (HPW and LPW) were selectively bred to display severe and mild pentobarbital withdrawal HICs, respectively. These mice provide a valuable means to assess genetic correlations between withdrawal from pentobarbital and other sedative agents. We tested HPW and LPW mice for severity of HICs elicited during withdrawal from ethanol, diazepam, and zolpidem, and measured consumption of and preference for pentobarbital solutions in HPW and LPW mice. HPW mice displayed greater HICs than LPW mice during ethanol and zolpidem withdrawal, but differed less robustly during diazepam withdrawal. LPW mice consumed more pentobarbital in a solution of a moderate concentration than did HPW mice, but did not consume more pentobarbital at a higher or lower concentration. These results indicate that some of the same genes that affect the severity of withdrawal from pentobarbital also influence ethanol and zolpidem withdrawal, but that diazepam withdrawal may be less influenced by these genes. PMID- 15120580 TI - Intracerebral xenografts of mouse bone marrow cells in adult rats facilitate restoration of cerebral blood flow and blood-brain barrier. AB - We examined in the present study alterations in cerebral blood flow (CBF) and blood-brain barrier (BBB) permeability following intrastriatal transplantation of mouse bone marrow stromal cells (BMSCs) or saline infusion in adult Sprague Dawley rats. Laser Doppler revealed that transplanted animals exhibited near normal cerebral blood flow (CBF, 150 perfusion units) at a much earlier period post-transplantation (day 4) compared to animals that received saline infusion (day 12) (p's<0.05). Similarly, Evans Blue assay demonstrated that transplanted animals exhibited near complete BBB reconstitution at day 5 post-transplantation, whereas animals that received saline infusion continued to display a compromised BBB up to 11 days post-transplantation. Transplanted animals displayed a cell dose-dependent CBF and BBB restoration. Enzyme-linked immunosorbent assay (ELISA) of transplanted BMSCs revealed elevated levels of transforming growth factor-beta superfamily of neurotrophic factors. Moreover, despite the absence of immunosuppression in this cross-species transplantation, at least in the acute phase (12 days post-transplantation), surviving xenografts were detected during periods of restored CBF and BBB permeability. These observations suggest that restoration of CBF and BBB permeability accompanies the reported functional outcomes associated with intracerebral transplantation of BMSCs. PMID- 15120581 TI - FK506 abrogates delayed neuronal death via suppression of nitric oxide production in rats. AB - BACKGROUND AND PURPOSE: The mechanism of the neuroprotective effect of FK506 in relation to nitric oxide (NO) production has not been clarified in vivo. We have investigated the effect of FK506 on ischemia-induced NO production in association with the pathogenesis of delayed neuronal death (DND) in rats. METHODS: In vivo microdialysis was performed in the hippocampus of male Sprague-Dawley rats (250 350 g). Dialysate samples were collected every 3 min. In the ischemia group (n=16), global ischemia was induced for 21 min and reperfusion was achieved. In the FK506 treatment group (n=25), FK506 (1 mg/kg, i.v.) was administered 21 min prior to the onset of global ischemia. Sham operations were done (n=15). The levels of NO(2)(-) in the dialysate samples were determined by the Griess reaction. The animals were decapitated 7 days after ischemia. Coronal brain sections were stained with hematoxylin and eosin. RESULTS: In the ischemia group, the NO(2)(-) level significantly increased during ischemia. In the FK506 treatment group, there was no significant change in the NO(2)(-) level during ischemia. In histological examinations, FK506 treatment showed a neuroprotective effect against DND. CONCLUSIONS: The effect of FK506 inhibiting NO production contributes to the neuro-protective effect of FK506 on DND in the hippocampus. PMID- 15120582 TI - Neuronal protection from glucose deprivation via modulation of glucose transport and inhibition of apoptosis: a role for the insulin-like growth factor system. AB - Glucose is the brain's major energy source; therefore, loss of neuronal cells is a potential consequence of hypoglycaemia. Since apoptosis is a major mechanism of neuronal loss following a range of insults, we explored potent anti-apoptotic systems (IGF-I and bcl-2) as means of enhancing neuronal survival in the face of glucose deprivation. Human neuroblastoma cells (SH-SY5Y, SHEP and SHEP-bcl-2) were exposed to low glucose as a model of glucopenia-induced neuronal damage. Administration of IGF-I and/or over-expression of the survival gene bcl-2 were exploited to attempt to limit neuronal loss. Neuronal survival mechanisms and interactions between these systems were investigated. Low glucose (0.25-2.5 mM) adversely affected cell growth and survival; however, IGF-I ameliorated these outcomes. Over-expression of bcl-2 blunted low glucose-induced apoptosis and up regulated IGF-I receptor, with the effect of IGF-I addition being negligible on apoptosis, while significantly enhancing mitochondrial activity. In SH-SY5Y cells, IGF-I significantly changed >two-fold mRNA levels of the apoptosis-related genes gadd45, fas, iNOS, NFkB, TRAIL, without further affecting bcl-2 expression. In low glucose, IGF-I acutely enhanced glucose transport and translocation of GLUT1 protein to the cell membrane. GLUT1 mRNA expression was up-regulated by both IGF-I and bcl-2. The potent anti-apoptotic systems IGF-I and bcl-2 are both thus able to enhance cell survival in a glucose-deprived human neuronal model. Although we clearly show evidence of positive cross-talk via bcl-2 modulation of IGF-I receptor, IGF-I also has enhancing effects on mitochondrial function outside the bcl-2 pathway. The common effect of both systems on enhancement of GLUT-1 expression suggests that this is a key mechanism for enhanced survival. These studies also point to the potential use of IGF-I therapy in prevention or amelioration of hypoglycaemic brain injury. PMID- 15120583 TI - Expression of MAP1B protein and its phosphorylated form MAP1B-P in the CNS of a continuously growing fish, the rainbow trout. AB - Microtubule-associated protein-1B (MAP1B), and particularly its phosphorylated isoform MAP1B-P, play an important role in axonal outgrowth during development of the mammalian nervous system and have also been shown to be associated with axonal plasticity in the adult. Here, we used antibodies and mRNA probes directed against mammalian MAP1B to extend our analysis to fish species, trout (Oncorhynchus mykiss), at different stages of development. The specificity of the cross-reaction of our anti-total-MAP1B/MAP1B-P antibodies was confirmed by Western blotting. Trout MAP1B-like proteins exhibited about the same apparent molecular weight (320 kDa) as rat-MAP1B. Immunohistochemistry and in situ hybridization analysis performed on hindbrain and spinal cord revealed the presence of MAP1B in neurons and some glial subpopulations. Primary sensory neurons and motoneurons maintain high levels of MAP1B expression from early stages throughout adulthood, as has been shown for mammals. Unlike mammals, however, MAP1B and axon-specific MAP1B-P continue to be strongly expressed by hindbrain neurons projecting into spinal cord, with the important exception of Mauthner cells. MAP1B/MAP1B-P immunostaining were also detected elsewhere within the brain, including axons of the retino-tectal projection. This obvious difference between adult fish and mammals is likely to reflect the capacity of fish for continued growth and regeneration. Our results suggest that MAP1B/MAP1B P expression is generally maintained in neurons known to regenerate after axotomy. The regenerative potential of the adult nervous system may in fact depend on continued expression of neuron-intrinsic growth related proteins, a feature of MAP1B that appears phylogenetically conserved. PMID- 15120584 TI - Effect of 1,2,3,4,-tetrahydroisoquinoline administration under conditions of CYP2D inhibition on dopamine metabolism, level of tyrosine hydroxylase protein and the binding of [3H]GBR 12,935 to dopamine transporter in the rat nigrostriatal, dopaminergic system. AB - Current concepts of Parkinson's disease (PD) postulate that interaction between neurotoxins and specific genetic background may play an important role in pathogenesis of PD. Therefore, the effect of multiple administration of 1,2,3,4 tetrahydroisoquinoline (TIQ) under conditions of CYP2D blockade on the expression of key markers of PD was studied in the rat striatum (STR) and substantia nigra (SN). TIQ administered alone (50 mg/kg i.p. twice daily for 14 days) markedly decreased the level of tyrosine hydroxylase protein (TH) in the STR; however, this effect was not accompanied by reduction of dopamine (DA) concentration and [(3)H]GBR 12,935 binding to dopamine transporter (DAT). Administration of CYP2D inhibitor, quinine, jointly with TIQ lowered the levels of TH and DA in that structure, but slightly increased DAT binding. In the SN, treatment with TIQ alone did not change TH level although it enhanced DA content and decreased [(3)H]GBR 12,935 binding to DAT in the substantia nigra pars compacta (SNc). Neither the TH level nor DA concentration was affected by the combined treatment, although DAT binding was still reduced in the SN. TIQ did not change the total DA catabolism in the STR, but caused its inhibition in the SN. It strongly depressed the levels of intraneuronal DA metabolite DOPAC and enhanced that of extraneuronal 3-MT in either structure. TIQ more weakly affected the levels of both DA metabolites in the presence of quinine. Our results suggest that endogenous TIQ may act rather as neuromodulator but not as parkinsonism-inducing neurotoxin in the rat brain. PMID- 15120585 TI - Blockade of GABAA receptors disrupts isoperiodic neuronal oscillations in the intergeniculate leaflet of the rat. AB - The intergeniculate leaflet of the thalamus is, besides the suprachiasmatic nucleus of the hypothalamus, the other important neuronal element of the mammalian biological clock. The extracellularly recorded activity of neurons constituting the intergeniculate leaflet, recorded in vivo, is characterized by distinct, very regular ultradian oscillations. The majority of neurons in the circadian timing system are GABAergic. Many, if not all, neurons of the suprachiasmatic nucleus and intergeniculate leaflet contain GABA. In the present study we examined the effects of the GABA(A) receptor antagonist bicuculline and the chloride channel blocker picrotoxin on isoperiodic neuronal oscillations in the intergeniculate leaflet of rats. We recorded extracellular multiple-unit neuronal activity from the intergeniculate leaflet of anesthetized rats. During the recording of isoperiodic oscillations, bicuculline or picrotoxin were stereotaxically injected at different concentrations into the lateral ventricle of rat brain. In all the experiments, injection of GABA(A) receptor antagonists transiently disrupted the isoperiodic phasic discharge recorded from the intergeniculate leaflet. These data suggest that GABA(A) receptors are involved in the generation of ultradian rhythmical neuronal oscillations in rat intergeniculate leaflet. PMID- 15120586 TI - Deleterious effects of an environmental noise on sleep and contribution of its physical components in a rat model. AB - Sleep disturbances induced by environmental noise (EN) exposure are now well admitted. However, many contradictory conclusions and discrepancies have been reported, resulting from uncontrolled human factors or the use of artificial noises (pure tone). Thus, the development of an animal model appears to be a useful strategy for determining whether EN is deleterious to sleep. The aims of this study were: (i) to confirm the effects of noise on sleep in a rat model; and (ii) to determine the most deleterious physical component of noise regarding sleep structure. For this purpose, rats were exposed during 24 h either to EN or to artificial broad-band noises [either continuous broad-band noise (CBBN) or intermittent broad-band noise (IBBN)]. All the noises decrease both slow wave sleep (SWS) and paradoxical sleep (PS) amounts during the first hours of exposure. However, CBBN acts indirectly on PS through a reduction of SWS bout duration, whereas IBBN and EN disturb directly and more strongly both SWS and PS. Finally, EN fragments SWS and decreases PS amount during the dark period, whereas IBBN only fragments PS. These results demonstrate the validity and suitability of a rodent model for studying the effects of noise on sleep and definitively show that sleep is disturbed by EN exposure. Two physical factors seem to be implicated: the intermittency and the frequency spectrum of the noise events, which both induce long-lasting sleep disturbances. An additive effect of frequency spectrum to intermittency tends to abolish all possible adaptations to EN exposure. Since sleep is involved in cognitive processes, such disturbances could lead to cognitive deficits. PMID- 15120587 TI - Bilateral organization of unilaterally generated activity in lumbar spinal motoneurons of the rat. AB - The spinal nucleus of the bulbocavernosus (SNB) is a medially located, bilaterally organized sexually dimorphic motor nucleus in the lumbar spinal cord of the male rat. To begin to assess the potential functional significance of this bilateral organization, we recorded ipsi- and contralateral SNB motor nerve activity following unilateral spinal stimulation and examined the timing, pattern, and recruitment of population motoneuron activity. A possible mechanism for bilateral communication, gap junctional intercellular communication, was also investigated because dye coupling experiments indicate an extensive syncytium in which SNB motoneurons are coupled with each other and neighboring interneurons. An in vivo peripheral nerve recording paradigm was used: a bipolar stimulating electrode was placed on dorsal root L6, and bipolar recording electrodes were placed bilaterally on the SNB motor nerves. All processes were severed distal to electrode placement to isolate the central preparation; recruitment curves of motoneuronal activity were then generated. Amplitude of peak to peak recruitment was greater in the contralateral motor nerve than in the ipsilateral nerve. Response latency, Fourier transform and spike counts showed no evidence of ipsi/contralateral asymmetry. Recruitment was attenuated both ipsi- and contralaterally after pharmacological gap junction blockade, but antidromic stimulation could not drive activity in contralateral motor axons. These results indicate that unilateral input to the SNB may be differentially modulated to produce functionally distinct output in the two separate halves of the nucleus. We also discuss the potential modulatory role of gap junctions in the activity of the SNB. PMID- 15120588 TI - Electrophysiological characterization of facilitated spinal withdrawal reflex to repetitive electrical stimuli and its modulation by central glutamate receptor in spinal anesthetized rats. AB - The present study is aimed to systematically investigate wind-up and after discharge of the spinal withdrawal reflex assessed by recording single motor unit (SMU) electromyographic (EMG) response to different intensities [0.5-1.5xreflex threshold (T)] of repetitive [frequencies (0.5-200 Hz)] transcutaneous electrical stimuli for 5 s. The role of central glutamate receptors in modulation of the withdrawal reflex facilitation was observed and evaluated in order to explore the potential central mechanism. Stimulus intensities below reflex threshold, such as 0.8xT, but not 0.5xT, could by repetition elicit and facilitate withdrawal reflex. The facilitation (wind-up and after-discharge) of the withdrawal reflex is a result of central integration and is increased significantly for increasing stimulus intensity and frequency. Electrical stimuli at 3-5 Hz for 5 s are appropriate to elicit wind-up. In contrast, 10-20 Hz frequencies of electrical stimuli are adequate to evoke the after-discharge. For pharmacological intervention, suprathreshold (1.5xT) repeated (5 Hz) electrically evoked facilitated reflex (wind-up) were apparently depressed by intrathecal (i.t.) administration of MK-801 as well as CNQX (40 nmol/10 microl, respectively). However, wind-up of spinal reflexes evoked by subthreshold (0.8xT) electrical stimuli could only be depressed by the treatment with CNQX, not MK-801. The after discharge of the withdrawal reflex elicited by 20 Hz electrical stimulation with either 0.8xT or 1.5xT intensity was depressed by i.t. treatment with CNQX. I.t. application of MK-801 only depressed 0.8xT the intensity of electrically evoked after-discharge. In conclusion, for the first time, the present study clearly demonstrates that, following the wind-up phase, the spinal withdrawal reflex pathways continue to fire spontaneously in a stimulus frequency- and intensity dependent way (temporal and/or spatial summation). This inherited memory and the central non-N-methyl-d-aspartate (non-NMDA) receptor, but not the NMDA receptor, mainly involving pharmacological mechanisms, may play an important role in pathological conditions with spontaneous nociceptive firing. Furthermore, the after-discharge of the spinal reflex may be an important indicator for studies on central sensitization in many pathological pain conditions. PMID- 15120589 TI - The use of stereological counting methods to assess immediate early gene immunoreactivity. AB - The issue of whether profile and stereological counting methods are interchangeably accurate when assessing immediate early gene expression still needs to be resolved. To compare these two counting techniques, we quantified the expression of c-fos in the nucleus accumbens core and shell, and in the lateral septum as a control structure, of rats treated with neuroleptics. With the profile counting method, which relies on selective placement of a counting grid within a structure, we evaluated the density of c-fos labeled cells within a box of fixed dimension. With stereology, which applies random and systematic sampling methods, we used the optical fractionator method and counted the absolute number of c-fos labeled cells within the contours of each structure examined. Our results showed that the substantial increase in c-fos expression in the shell and core induced by haloperidol treatment was detected by both stereological and profile counting methods; in contrast, the weaker effect of clozapine on c-fos expression was detected differentially by the two methods. Whereas the profile counting method reported a reduction of c-fos in the core by clozapine, and an increase in c-fos in the lateral septum, these effects were not replicated using stereology. These findings suggest that stereological and profile counting methods do not always produce equivalent results. This may be particularly relevant when a measured effect is relatively small, and it is not distributed homogeneously within a structure. In this respect, the random and systematic sampling methods of stereology may yield more accurate and unbiased results than the profile counting method, and therefore may be preferred for a more accurate and thorough investigation of a treatment effect on immediate early gene expression in a specific brain region. PMID- 15120590 TI - State-specific asymmetries in EEG slow wave activity induced by local application of TNFalpha. AB - Sleep is posited to be a fundamental property of groups of highly interconnected neurons and regulated in part by activity-dependent sleep regulatory substances such as tumor necrosis factor alpha (TNFalpha). We show that the unilateral local application of TNFalpha onto the somatosensory cortex of rats induced state- and frequency-dependent EEG asymmetries. In contrast, the unilateral injection of a TNFalpha inhibitor, a TNFalpha soluble receptor, attenuated sleep deprivation enhanced EEG slow wave power ipsilaterally during non-rapid eye movement sleep (NREMS) but not during REMS or waking. Results are consistent with the notion that sleep begins with state changes occurring within small groups of highly interconnected neurons and is driven in part by the local production of sleep regulating substances. PMID- 15120591 TI - Target site of inhibition of baroreflex vagal bradycardia by nasal stimulation. AB - We have previously reported that stimulation of nasal mucosa inhibits baroreflex vagal bradycardia (BVB) and this inhibition was mediated exclusively by the trigeminal nerve, and occurred principally at pontomedullary level. In this study, to identify the target site of the inhibition, several types of experiments were conducted in chloralose-urethane-anesthetized, beta-adrenergic receptor-blocked rats. Afferent discharges in the ethmoidal nerve (EN5) were increased in response to nasal stimulation by smoke, and electrical stimulation of the EN5 suppressed BVB induced by electrical stimulation of the aortic depressor nerve (ADN). Electrical stimulation of the EN5 inhibited vagal bradycardia evoked by either electrical or chemical stimulation of the nucleus tractus solitarius (NTS), while it rather facilitated bradycardia by stimulation of the nucleus ambiguus (NA) region. Microstimulation of the NTS induced antidromic compound spike potential along the ADN but this was not affected by stimulation of the EN5. ADN-evoked field potentials and unitary responses of neurons in the NTS were suppressed by stimulation of the EN5. These results suggested that barosensitive neurons in the NTS are the major target sites of inhibition of BVB by nasal stimulation in rats. PMID- 15120592 TI - Differential action potentials and firing patterns in injured and uninjured small dorsal root ganglion neurons after nerve injury. AB - The profile of tetrodotoxin sensitive (TTX-S) and resistant (TTX-R) Na(+) channels and their contribution to action potentials and firing patterns were studied in isolated small dorsal root ganglion (DRG) neurons after L5/L6 spinal nerve ligation (SNL). Total TTX-R Na(+) currents and Na(v) 1.8 mRNA were reduced in injured L5 DRG neurons 14 days after SNL. In contrast, TTX-R Na(+)currents and Na(v) 1.8 mRNA were upregulated in uninjured L4 DRG neurons after SNL. Voltage dependent inactivation of TTX-R Na(+) channels in these neurons was shifted to hyperpolarized potentials by 4 mV. Two types of neurons were identified in injured L5 DRG neurons after SNL. Type I neurons (57%) had significantly lower threshold but exhibited normal resting membrane potential (RMP) and action potential amplitude. Type II neurons (43%) had significantly smaller action potential amplitude but retained similar RMP and threshold to those from sham rats. None of the injured neurons could generate repetitive firing. In the presence of TTX, only 26% of injured neurons could generate action potentials that had smaller amplitude, higher threshold, and higher rheobase compared with sham rats. In contrast, action potentials and firing patterns in uninjured L4 DRG neurons after SNL, in the presence or absence of TTX, were not affected. These results suggest that TTX-R Na(+) channels play important roles in regulating action potentials and firing patterns in small DRG neurons and that downregulation in injured neurons and upregulation in uninjured neurons confer differential roles in shaping electrogenesis, and perhaps pain transmission, in these neurons. PMID- 15120593 TI - Altered Bad localization and interaction between Bad and Bcl-xL in the hippocampus after transient global ischemia. AB - Accumulating evidence indicates that the mitochondrial cell-death pathway, which involves the release of cytochrome c from mitochondria, participates in neuronal cell death after transient cerebral ischemia. However, the upstream events, that induce cytochrome c release after transient global ischemia are not fully understood. Bad is a pro-apoptotic member of the bcl-2 gene family that promotes apoptosis by binding to and inhibiting functions of anti-apoptotic proteins Bcl-2 and Bcl-xL. We investigated the effects of transient (15 min) global ischemia on the intracellular localization of Bad and the interaction of Bad with calcineurin, Akt or Bcl-xL in the vulnerable CA1 and resistant CA3/dentate gyrus of the hippocampus. Immunoblotting analysis revealed that the amount of Bad in mitochondria significantly increased after ischemia. Co-immunoprecipitation studies showed decreased interactions of Bad with Akt and calcineurin in the cytosol and increased binding with Bcl-xL in the mitochondrial fraction of hippocampal CA1, but not in the CA3/dentate gyrus region. Further, we examined the effect of recombinant Bad on the cytochrome c release from isolated mitochondria. Treatment with both recombinant Bad and calcium, but not with recombinant Bad alone, induced cytochrome c release. These results suggest that changes in localization and complex formation by Bad are, at least in part, involved in the vulnerability of cells after transient global ischemia. PMID- 15120594 TI - Serotonin 5-HT2 and 5-HT1A receptors in the periaqueductal gray matter differentially modulate tonic immobility in guinea pig. AB - Tonic immobility (TI) is an inborn defensive behavior characterized by a temporary state of profound and reversible motor inhibition elicited by some forms of physical restraint. We have previously reported that cholinergic stimulation of the dorsal periaqueductal gray matter (PAG) decreases the duration of TI episodes, while stimulation of the ventrolateral region increases it. The ventrolateral PAG modulates this behavior via a similar neural circuit proposed to be involved in the antinociceptive system. Some studies have indicated that alterations in the levels of cerebral 5-hydroxytryptamine (5-HT) mediate or modulate the analgesic effect of PAG stimulation. Thus, in this study we investigated the possibility that the serotoninergic system is involved in the modulation of TI by this neural substrate. Our results showed that the effect of serotonin into the ventrolateral and dorsal PAG seems to be biphasic and dose dependent. The microinjection of low doses (0.1 microg) of 5-HT into the PAG increased the duration of TI, while high doses (1, 3 and 6 microg) decreased this behavior. Our results also showed that microinjection of a 5-HT(1A) agonist (0.003, 0.01 and 0.1 microg of 8-hydroxy-dipropylaminotretalin (8-OH-DPAT)) into the PAG increased the duration of TI episodes. However, the microinjection of 5 HT(2) agonist (0.01 and 0.1 microg of alpha-methyl-5-HT) into the PAG decreased the duration of TI and this effect could be reversed by pretreatment with an ineffective dose (0.01 microg) of ketanserin. In contrast, ketanserin (0.03 and 0.16 microg) increased this behavior in a dose-dependent manner. These results suggest that the PAG 5-HT(1A) and 5-HT(2) receptors have different roles in the modulation of TI in guinea pigs, since the 5-HT(1A) and 5-HT(2) agonists, respectively, increased and decreased the duration of TI. PMID- 15120596 TI - Hippocampal dysfunction in Gulf War Syndrome. A proton MR spectroscopy study. AB - The pathogenesis of Gulf War Syndrome (GWS) is not clearly understood. Data exist to suggest that GWS may originate from a combination of chronic fatigue and sensitivity to the exposure of exogenous agents. Since the head region of hippocampus is highly vascularized and thus vulnerable to toxic substances in circulation, we postulated that hippocampal impairment occurs in GWS. To test this, single volume localized in vivo proton MR spectroscopy (MRS) studies of the left and right hippocampi of consenting Gulf War veterans (N=15; 10 with GWS, and 5 without GWS) and control Vietnam veterans (N=6) were conducted in accordance with approved human study protocols. The N-acetyl aspartate (NAA) to creatine and choline to creatine ratios were computed from the spectra. The NAA/creatine ratio of the GWS group (N=10) was found to be significantly lower than that of the entire control group (N=11) or the unaffected GW control group (N=5). No laterality differences were observed among any of the three groups. The choline/creatine ratio of the GWS group was not different from that for either control group. To check the existence of any relationship between age and the NAA/creatine ratios, the entire study population was grouped into those below or above the median age (44.3 years). It was found that the NAA/Cre ratio of the younger group (only Gulf War veterans) was significantly lower than that of the older group. The lower NAA/creatine ratio for the GWS group points to the existence of hippocampal dysfunction. PMID- 15120595 TI - Neural pathways involved in food allergy signaling in the mouse brain: role of capsaicin-sensitive afferents. AB - There is increasing evidence supporting the notion that brain-gut communication is crucial for the manifestation of functional gastrointestinal (GI) disorders. Employing denervation by neonatal capsaicin treatment, we investigated here the role of unmyelinated C-fibers in food allergy signaling in the brain. We found that 90 min after oral ovalbumin (OVA) challenge, allergic mice present increased c-fos expression in emotionality-related brain areas such as the paraventricular nucleus of the hypothalamus (PVN) and the central nucleus of the amygdala (CeA). Food allergy also induced enhanced Fos immunoreactivity in the nucleus of tractus solitarii (NTS) of OVA-immunized animals. We also show that while the degree of Fos staining in the NTS of allergic mice was only diminished by neonatal capsaicin, it was completely blocked in the PVN. However, capsaicin did not modify food allergy-induced c-fos expression in the CeA. In conclusion, this study provides evidence showing that unmyelinated C-fibers are part of the neural pathways involved in food allergy-induced activation of specific brain areas, particularly the PVN and to a lesser extent the NTS. PMID- 15120597 TI - Effect of protein kinases on lactate dehydrogenase activity in cortical neurons during hypoxia. AB - Our previous work shows that delta-opioid receptor (DOR) protects cortical neurons from hypoxic insults. Since an enhanced anaerobic capacity is important for neurons to adapt to the reduction of oxidative phosphorylation, we asked whether DOR plays a role in neuronal regulation of anaerobic capacity, thus protecting neurons from O(2) deprivation. Indeed, there is evidence suggesting that DOR may regulate protein kinase A (PKA) and C (PKC), which are involved in regulation of lactate dehydrogenase (LDH). However, little is known regarding the role of DOR and protein kinases in the regulation of glycolytic and related enzymes. As a first step, the present studies were performed in primary cultures of rat cortical neurons to clarify two issues: (1) Are protein kinases involved in the regulation of LDH activity in hypoxia? and (2) Does DOR affect LDH activity in hypoxic neurons? The results showed that PKC activation yielded substantial increases in normoxic LDH activity and significantly augmented LDH activity in hypoxic neurons, while PKC inhibition decreased LDH activity in both normoxic and hypoxic neurons. PKA activation significantly increased LDH activity in normoxic neurons and further elevated LDH activity in hypoxic neurons. However, PKA inhibition did not decrease in LDH activity in either normoxic or hypoxic neurons. Although DOR inhibition slightly reduced LDH activity in normoxia, DOR activation or inactivation did not alter LDH activity in hypoxic neurons. These data suggest that in cortical neurons, (i) PKC up-regulates LDH activity and plays an important role in its up-regulation during hypoxia; (ii) PKA is less likely involved in the regulation of LDH activity during hypoxia although its stimulation may slightly increase LDH activity and (iii) DOR does not contribute to LDH activity up-regulation during hypoxia. PMID- 15120599 TI - Different levels of neuroprotection by two insulin-like growth factor-I splice variants. AB - We compared the neuroprotective effects of a liver-type isoform of insulin-like growth factor-I (IGF-IEa) and its splice variant, mechano-growth factor (MGF), isolated from active skeletal muscle. cDNAs of these peptides were injected into the facial muscle of adult rats prior to facial nerve avulsion. This resulted in significant neuroprotection of 88% and 37%, respectively, of motoneurons compared to control plasmid and avulsion-only groups. MGF is markedly more effective than the liver-type, systemic IGF-I for motoneuron survival, suggesting a major role for the peripheral target in adult neuronal maintenance and survival. PMID- 15120598 TI - The role of superoxide dismutase and alpha-tocopherol in the development of seizures and kindling induced by pentylenetetrazol - influence of the radical scavenger alpha-phenyl-N-tert-butyl nitrone. AB - Previous experiments have shown that the generation of free hydroxyl radicals in rat brain homogenates is increased following pentylenetetrazol (PTZ) kindling. The present study was performed in order to evaluate the involvement of endogeneous radical defence systems as the superoxide dismutase (SOD) and the level of alpha-tocopherol, an important lipid-soluble and membrane-bound antioxidant in brain homogenate of rats after acute seizure and kindling induced by PTZ. The activities of the total SOD were significantly reduced after acute seizure and tend towards an enhancement in kindled animals. Western blot analysis shows an upregulation of Mn-SOD in rat brain homogenates after kindling. The level of the chain-breaking antioxidant alpha-tocopherol was reduced in acutely convulsing rats and was not modified in kindled rats. Second, we studied the influence of exogeneously supplied radical scavenger alpha-phenyl-N-tert-butyl nitrone (PBN) on seizure and kindling following PTZ treatment. After a single injection of PTZ at a dose evoking clonic-tonic seizures, PBN did not modify either the formation of free hydroxyl radicals measured by the levels of 2,3 dihydroxybenzoic acid (DHBA) and 2,5-DHBA or the susceptibility to PTZ. In the kindling group, subchronic treatment with PBN (over a period of 4 weeks) prevented the increase in the formation of free hydroxyl radicals, and the susceptibility to PTZ was transiently decreased during the development of kindling, but PBN did not influence the susceptibility to PTZ in fully kindled rats. Pretreatment with PBN increased the activities of total SOD and the protein content of Mn-SOD and decreased the level of alpha-tocopherol in comparison to saline controls. The results suggest that the formation of free hydroxyl radicals is not reflected by an enhanced susceptibility to PTZ classified according to the modified RACINE scale. Additionally, it may be assumed that the increased generation of hydroxyl radicals in kindled animals is not primary caused by an exhaustion of both the defence systems measured. Adaptive mechanisms, as the induction of Mn-SOD, may be taken into consideration to counteract oxidative stress-mediated free radical formation. PMID- 15120600 TI - Postmortem effect of pentobarbital anesthetic on survival of adult cortical neurons in primary culture. AB - We determined whether pentobarbital anesthetic is required to culture postmortem adult rat neurons. Pentobarbital treatment resulted in two-fold increases in neuron survival in culture after 2 and 4 h postmortem compared to non-anesthetic controls, but was not as effective as simple postmortem treatment on ice and therefore not essential. PMID- 15120601 TI - Lesions of the periaqueductal gray disrupt input to the rostral ventromedial medulla following microinjections of morphine into the medial or basolateral nuclei of the amygdala. AB - Microinjections of morphine into the basolateral (BLa) and medial (MEa) nuclei of the amygdala differentially affect rostral ventromedial medulla (RVM) neuronal activity and nocifensive behaviors. PAG lesions attenuated or blocked the effects of both BLa and MEa morphine on RVM cell activity, and interfered with the behavioral antinociception produced by BLa infusions. These results demonstrate that the influences from both the BLa and MEa to the RVM are relayed via the PAG. PMID- 15120602 TI - Prenatal cocaine exposure decreases brain-derived neurotrophic factor proteins in the rat brain. AB - The pregnant rats received daily sc injections of cocaine (30 mg/kg) or saline from the gestational day (GD) 7 to GD 20. At 1 week postnatal, all pups were killed and the hippocampus, cortex and striatum were dissected out. Levels of brain-derived neurotrophic factor (BDNF) under the basal condition and depolarization with high potassium (40 mM) were measured. The results showed that hippocampal BDNF levels under basal and depolarization conditions were all significantly lower in the pups prenatally exposed to cocaine than those exposed to saline. There were no significant differences in basal BDNF levels between the cocaine and saline groups in the cortex or striatum. However, the prenatally cocaine-treated pups showed significantly less BDNF release following high potassium depolarization than the saline-treated animals did in both these regions. The results support the suggestion that prenatal cocaine exposure decreases BDNF expression in the offspring. PMID- 15120603 TI - Excitatory effects of ATP on rat dorsomedial hypothalamic neurons. AB - We investigated P2X purinoceptors in rat dorsomedial hypothalamic (DMH) neurons using nystatin-perforated patch-clamp recordings and fura-2 microfluorometry. Adenosine triphosphate (ATP) concentration-dependently evoked an inward current and increased cytosolic Ca(2+) ([Ca](i)). The rise in [Ca](i) was dependent on external Ca(2+) and Na(+), was blocked by Ca(2+) channel antagonists and had pharmacological properties consistent with P2X2 receptors. These results suggest that P2X receptor-mediated depolarization activates voltage-gated Ca(2+) channels, resulting in an increase in [Ca](i). PMID- 15120604 TI - MK-0767, a novel dual PPARalpha/gamma agonist, displays robust antihyperglycemic and hypolipidemic activities. AB - Here, we characterize the actions of MK-0767, a dual ligand of the nuclear receptors peroxisome proliferator-activated receptor (PPAR)alpha and PPARgamma. In cell-based assays, MK-0767 produced potent activation of human PPARgamma and PPARalpha with a gamma:alpha potency ratio of approximately 2. The dual agonist induced high affinity interactions of PPARalpha and PPARgamma with the transcriptional coactivator CBP in vitro. In ob/ob mice, MK-0767 normalized hyperglycemia and hyperinsulinemia with equal or greater potency and efficacy than pioglitazone. Treatment of hamsters with MK-0767 produced substantial reductions in blood cholesterol and triglycerides. In dogs, MK-0767 reduced serum cholesterol levels with a potency more than 10-fold greater than simvastatin. The efficacies of MK-0767 and simvastatin were additive when given together. We conclude that MK-0767 is a potent dual PPARalpha/gamma agonist with robust insulin sensitizing and hypolipidemic activities. PMID- 15120605 TI - PPARalpha activation abolishes LDL-stimulated IL-8 production via AP-1 deactivation in human aortic smooth muscle cells. AB - Native low density lipoprotein (n-LDL) is a major risk factor for cardiovascular diseases by inducing inflammatory processes and vascular smooth muscle cell proliferation in vessel cells. It has previously been reported that LDL enhances inflammatory reactions by the up-regulation of interleukin (IL)-8 via the activation of p38 kinase and activator protein (AP)-1 in human aortic smooth muscle cells (hAoSMCs). The findings of this study show, for the first time, that the peroxisome proliferator-activated receptor (PPARalpha) agonist, fenofibrate, completely abolishes the LDL-induced IL-8 up-regulation at the transcriptional level. Pretreatment of hAoSMCs with fenofibrate abolishes the effects of LDL on AP-1 activation without affecting nuclear factor (NF)-kappaB. In contrast, fenofibrate failed to modulate the activation state of p38 and JNK kinases or the levels of c-fos and phospho-Jun. These data suggest that AP-1 is likely to be located at the crossroads between LDL signaling and the regulation of IL-8 modulation by PPARalpha. PMID- 15120606 TI - A novel 3-step enantioselective synthesis of pyrenylalanine with subsequent incorporation into opioid, CCK, and melanotropin ligands. AB - Pyrene possesses unique spectroscopic properties such as a high quantum yield, a long half-life in the excited state, and the ability to form excimers when in proximity to each other in the excited state. These properties allow pyrenylalanine, which is a pyrene moiety incorporated into an amino acid, to be used as a fluorescent probe in peptides and proteins. The common route for the synthesis of pyrenylalanine involves 5 steps, with subsequent separation of the two isomers by recrystallization. This paper reports a novel 3-step asymmetric synthesis of pyrenylalanine with high enantioselectivity, good yields, and facile isomer purification. After synthesis, pyrenylalanine was incorporated into a series of opioid, CCK, and melanotropin peptide ligands in order to study the effects of aromaticity, lipophilicity, and steric properties on their potency and efficacy at their corresponding biological receptors. The change in binding and efficacy of the labeled ligands as compared to the unlabeled ligands demonstrates the possible role of lipophilicity/aromaticity in the binding and signal transduction of the ligand-receptor interaction. PMID- 15120607 TI - Interference of hepatitis A virus replication by small interfering RNAs. AB - The rate of acute liver failure due to hepatitis A virus (HAV) has not decreased, and therapy of severe infections is still of major interest. Using a DNA-based HAV replicon cell culture system, we demonstrate that small interfering RNAs (siRNAs) targeted against viral sequences or a reporter gene contained in the viral genome specifically inhibit HAV RNA replication in HuhT7 cells. Combinations of siRNAs were more effective suppressors of HAV RNA replication. Also, siRNAs targeted against HAV 2C and 3D inhibited the expression of the respective protein. Expressions of endogenous beta-actin and double-stranded specific RNA-activated serin/threonine kinase (PKR) were unaltered, demonstrating that the siRNA inhibitory effect was not connected to interferon inhibition, but rather was specifically targeted against HAV RNA. These results suggest that RNA interference might ultimately be useful in treatment of severe HAV infection with or without chronic liver diseases. PMID- 15120608 TI - Carboplatin induces apoptotic cell death through downregulation of constitutively active nuclear factor-kappaB in human HPV-18 E6-positive HEp-2 cells. AB - Because the role of nuclear factor kappaB (NF-kappaB) is in cellular growth control and neoplasia, we explored the status of NF-kappaB and investigated its role in survival of human HPV-18 E6-positive HEp-2 cells. We observed accumulation of p65 in the nucleus. Moreover, without any external stimulus constitutive NF-kappaB DNA binding and transactivation activity were detected in HEp-2 cells. Treatment with NF-kappaB inhibitor curcumin (diferuloylmethane) and transient transfection of the mutant form of IkappaBalpha, IkappaBalpha super repressor (IkappaBalpha-SR), suppressed constitutive NF-kappaB activity as well as proliferation, suggesting that constitutive NF-kappaB activity is required for the survival of HEp-2 cells. Carboplatin treatment downregulated constitutive NF kappaB activity and prevented nuclear retention of p65. Further, carboplatin also suppressed the constitutive IkappaBalpha phosphorylation leading to stabilization of IkappaBalpha protein in the cells. Carboplatin inhibited NF-kappaB binding to its response element present in Bcl-2 promoter resulting in downregulation of antiapoptotic Bcl-2 protein. Thus, our results for the first time indicate that constitutive NF-kappaB has a significant role in the survival of HPV-18 E6 positive HEp-2 cells. Moreover, inactivation of NF-kappaB is one of the mechanisms underlying the induction of carboplatin-mediated apoptosis in HPV-18 E6-positive cancer cells. PMID- 15120609 TI - The C-terminal domain of canstatin suppresses in vivo tumor growth associated with proliferation of endothelial cells. AB - Angiogenesis is crucial for the growth and metastasis of solid tumors with sizes larger than a few cubic millimeter Canstatin, the non-collagenous 1 (NC1) domain of alpha2 chain of type IV collagen, was previously shown to inhibit proliferation of endothelial cells in vitro and suppress in vivo tumor growth. Our previous studies showed that canstatin-N, the N-terminal 1-89 amino acid fragment of canstatin, inhibited the neovascularization in vivo, potently induced apoptosis of endothelial cells in vitro, and suppressed in vivo tumor growth in BALB/c mice. In the present study, we demonstrated that canstatin-C, the C terminal 157-227 amino acid fragment of canstatin, also specifically inhibited in vitro the proliferation of human umbilical vein endothelial cells and induced apoptosis, but the apoptosis-inducing activity, while close to that of the full length canstatin, was much lower than that of canstatin-N. Canstatin-C also suppressed in vivo tumor growth in BALB/c mice at a dosage of 10mg/kg/day. These results suggest that canstatin-C is an anti-angiogenic domain of canstatin mainly associated with the specific inhibition of proliferation of endothelial cells, whereas canstatin-N with the potential apoptosis-inducing activity on endothelial cells. PMID- 15120610 TI - The human involucrin gene is transcriptionally repressed through a tissue specific silencer element recognized by Oct-2. AB - Involucrin is an important marker of epithelial differentiation which expression is upregulated just after basal cells are pushed into the suprabasal layer in stratified epithelia. Several transcription factors and regulatory elements had been described as responsible for turning on the gene. However, it is evident that in basal cell layer, additional mechanisms are involved in keeping the gene silent before the differentiation process starts. In this work, we located a potential transcriptional silencer in a 52bp sequence whose integrity is necessary for silencing the proximal enhancer promoter element (PEP) in multiplying keratinocytes. Octamer-binding sites were noticed in this fragment and the specific binding of Oct-2 transcription factor was detected. Oct-2 appears to be implicated in an epithelial-specific repression activity recorded only in keratinocytes and C33-A cell line. Overexpression of Oct-2 repressed the involucrin promoter activity in epithelial cells and in the presence of the silencer element. PMID- 15120611 TI - 5-Aminoimidazole-4-carboxamide riboside suppresses lipopolysaccharide-induced TNF alpha production through inhibition of phosphatidylinositol 3-kinase/Akt activation in RAW 264.7 murine macrophages. AB - 5-Aminoimidazole-4-carboxamide riboside (AICAR) is an adenosine analog and a widely used activator of AMP-activated protein kinase (AMPK). We examined the effect of AICAR on LPS-induced TNF-alpha production in RAW 264.7 and peritoneal macrophages and its molecular mechanism in RAW 264.7 macrophages. Treatment with AICAR inhibited LPS-induced increases in TNF-alpha mRNA and protein levels in these cells. AICAR or LPS did not alter the AMPK activity as well as the phosphorylations of AMPK alpha (Thr172) and ACC (Ser79). Moreover, an adenosine kinase inhibitor 5'-iodotubercidin enhanced the suppressive effect of AICAR on TNF-alpha levels. These results suggest that the effect of AICAR on TNF-alpha suppression in RAW 264.7 cells is independent of AMPK activation. In addition, an adenosine receptor antagonist 8-SPT had no effect on AICAR-induced suppression of TNF-alpha levels. Finally, we observed that AICAR inhibited LPS-induced activation of PI 3-kinase and Akt, whereas it had no effect on the activation of p38 and ERK1/2. Taken together, these results suggest that the anti-inflammatory action of AICAR in RAW 264.7 macrophages is independent of AMPK activation and is associated with inhibition of LPS-induced activation of PI 3-kinase/Akt pathway. PMID- 15120612 TI - Expression and secretion of human glucocerebrosidase mediated by recombinant lentivirus vectors in vitro and in vivo: implications for gene therapy of Gaucher disease. AB - Gaucher disease is a lysosomal storage disorder resulting from a deficiency of glucocerebrosidase (GC). In this study, we showed that vascular and hepatic delivery of a HIV-1-based lentivirus vector encoding human GC cDNA produced therapeutic levels of GC protein. A high level of expression of GC was produced in cultured fibroblasts derived from patients with Gaucher disease by transducing the cells with recombinant lentivirus vectors. GC secreted by transduced fibroblasts was taken up by adjacent GC-deficient cells by endocytosis. Intraportal administration of lenti-EF-GC viral vector resulted in efficient transduction and expression of the GC. Vascular delivery of vector resulted in high levels of GC expression in mice that persisted in most organs over the four months. No significant abnormalities were found attributable to recombinant lentivirus vectors in any of the tissues examined. This study represents an initial step toward gene transfer using recombinant lentivirus vectors for treatment of Gaucher disease. PMID- 15120613 TI - Human programmed cell death 5 protein has a helical-core and two dissociated structural regions. AB - Programmed cell death 5 (PDCD5) protein is phylogenetically conserved in both the nucleus and cytoplasm. The human PDCD5 protein is expressed in tumor cells during apoptosis independent of the apoptosis-inducing stimuli, and recently it was found that PDCD5 is an important regulator in both apoptotic and non-apoptotic programmed cell death. In this study, human PDCD5 was expressed in Escherichia coli cell and studied using heteronuclear NMR method. The NMR results indicate that PDCD5 protein can be divided into three structural regions, a core region and two dissociated terminal regions. The core region (41-101) represents a rigid sub-domain consisting mainly of a triple-helix bundle. The N-terminal 38 residues (3-40) are ordered, but not a rigid structural region which contains abundant secondary structure, and packs very loosely against the core. The C-terminal 17 residues (102-118) represent a mobile unstructured region, which may be capable of interaction with nucleic acid. PMID- 15120614 TI - Cooperativity in the binding of the cationic biocide polyhexamethylene biguanide to nucleic acids. AB - The interaction between the broad-spectrum antimicrobial agent, polyhexamethylene biguanide (PHMB), and various nucleic acids was investigated. Titration of either single- or double-stranded 100-bp DNA, or mixed-molecular weight marker DNA, or tRNA with PHMB caused precipitation of a complex between nucleic acid and PHMB in which the nucleotide/biguanide ratio was always close to unity. Binding of PHMB was highly cooperative, with apparent Hill coefficients 10.3-14.6. When a fluorescent derivative of PHMB was titrated with increasing amounts of nucleic acid, all four forms of nucleic acid caused strong polarisation of fluorescence, demonstrating the association with PHMB. The intensity and broad-spectrum binding of PHMB to all forms of nucleic acid has significant implications for the mechanism of action of this biocide. PMID- 15120615 TI - Nitrosylhemoglobin formation after infusion of NO solutions: ESR studies in pigs. AB - A saturated nitric oxide (NO) solution (1.88 mM) infused i.v. in the anesthetized pig at a dose of 68 nmol/kg/min for 24 min resulted in a time-dependent increase of nitrosylhemoglobin [HbFe(II)NO] as determined by electron spin resonance (ESR), reaching a C(max) of 7.99 +/- 0.42 microM at the end of the infusion, compared to 1.13 +/- 0.42 microM before (p < 0.01). This indicates that NO i.v. is efficiently bioconserved as HbFe(II)NO (approximately 34% of the NO dose) and to a greater extent than by the oxidative pathway (approximately 24% of the NO dose), as determined by measuring plasma nitrites/nitrates (chemiluminescence) and Met-Hb (ESR analysis). When the NO infusion was stopped, HbFe(II)NO declined with a t(1/2) of 15 min, indicating that it is a stable storage form of NO, able to deliver NO distally to the site of administration. No significant differences were observed in systemic and pulmonary vascular resistances during and after NO infusion, but PO(2) showed a significant decrease 15 and 30 min after the infusion. Thus, in normoxic/physiological conditions, HbFe(II)NO does not induce significant NO-dependent vasorelaxation. PMID- 15120616 TI - betaPix-b(L), a novel isoform of betaPix, is generated by alternative translation. AB - betaPix (Pak-interacting exchange factor) isoforms are recently identified guanine nucleotide exchange factors (GEFs) for Rho family GTPases, Rac/Cdc42, that are key players in the regulation of actin dynamics. Here we show that a novel 105-kDa betaPix isoform, betaPix-bL, is generated by alternative translation of betaPix-b mRNA. Translation of betaPix-bL starts at an atypical initiation site, GTG, that is located 57 nucleotides downstream from the newly identified 5' end of betaPix-b cDNA. The expression of two isoforms, betaPix-b and betaPix-bL, from betaPix-b mRNA is controlled by an internal ribosome entry site (IRES)-driven mechanism. Comparing to betaPix-b, betaPix-bL contains additional 105 amino acids composed of a calponin homology (CH) domain and a serine-rich sequence in the N-terminus. The expression of betaPix-bL in rat brain is developmentally regulated and high in the embryonic stages, suggesting that the function of betaPix-bL is more heavily required during the early stages of brain development. PMID- 15120618 TI - Expression and induction of CYP3As in human fetal hepatocytes. AB - CYP3A4 and CYP3A7 mRNA expression levels were markedly up-regulated by dexamethasone (DEX), but not by rifampicin (RIF). CYP3A5 mRNA level was not increased significantly by DEX, RIF, or phenobarbital. Testosterone 6beta hydroxylase activity was induced to about 2-fold of control by DEX. However, concomitant treatment with RIF did not alter DEX-mediated induction of CYP3A mRNA expression and testosterone 6beta-hydroxylase activity. DEX-mediated induction of CYP3A mRNA was suppressed in a dose-dependent manner by RU486, a glucocorticoid receptor (GR) antagonist. At 5microM RU486, DEX-mediated induction of CYP3A4, CYP3A5, and CYP3A7 mRNA expression was inhibited almost completely. These results suggest that, in human fetal hepatocytes, PXR is not involved in DEX-mediated induction of CYP3A4 and CYP3A7, and that the induction is mediated directly by GR. PMID- 15120617 TI - TWEAK/Fn14 interaction stimulates human bronchial epithelial cells to produce IL 8 and GM-CSF. AB - TNF-like weak inducer of apoptosis (TWEAK), a member of the tumor necrosis factor (TNF) family, is a multifunctional cytokine that regulates cellular proliferation, angiogenesis, inflammation, and apoptosis. In this study, we investigated the effect of TWEAK on human bronchial epithelial cells. A human bronchial epithelial cell line, BEAS2B, expressed a TWEAK receptor, fibroblast growth factor-inducible 14 (Fn14), and produced IL-8 and GM-CSF upon TWEAK stimulation in a dose-dependent manner, which was abrogated by anti-Fn14 blocking antibody. TWEAK induced phosphorylation of IkappaBalpha and BAY11-7082, a selective inhibitor of IkappaBalpha phosphorylation, inhibited the TWEAK-induced IL-8 and GM-CSF production by BEAS2B cells. Moreover, primary cultured human bronchial epithelial cells also expressed Fn14 and produced IL-8 and GM-CSF upon TWEAK stimulation. Collectively, TWEAK stimulated human bronchial epithelial cells to produce IL-8 and GM-CSF through Fn14. Because IL-8 and GM-CSF are associated with inflammatory conditions, these results suggest that TWEAK/Fn14 interaction may play some roles in airway inflammatory responses. PMID- 15120619 TI - Novel function of PS2V: change in conformation of tau proteins. AB - Neurofibrillary tangles (NFTs) are associated with many neurodegenerative disorders, such as Alzheimer's disease (AD). The major components of NFTs are hyper-phosphorylated tau proteins. The alternatively spliced form of the presenilin-2 (PS2) gene (PS2V) has been observed in sporadic AD brains. However, it is not known whether there is a relationship between tau aggregation/hyper phosphorylation and PS2V expression. In this manuscript, we make the first report of PS2V alterations in the conformation of the tau protein (unknown form of tau) in the human neuroblastoma cell line. PMID- 15120620 TI - Zyxin is up-regulated during megakaryocytic differentiation of human UT-7/c-mpl cells. AB - To characterize genes involved in megakaryocytic commitment, we compared expression profiles of bipotent cells (UT-7/c-mpl) with those of the same cells induced to differentiate towards megakaryopoiesis in the presence of TPO. Using cDNA arrays, we showed that 12 out of 2260 genes changed their expression level after 6h of TPO stimulation. One of these genes encodes for zyxin, a cytoskeleton protein component. Zyxin is up-regulated at the mRNA and protein levels in UT-7/c mpl cells in response to TPO confirming the reliability of the cDNA array technology. Similarly, when CD34 positive cells were induced to differentiate into megakaryocytes, zyxin mRNA was accumulated. Furthermore, when megakaryocytes were allowed to spread on fibrinogen, formation of stress fibers and lamellipodia was induced and zyxin was localized at the picks of actin stress fibers. These results suggest an important role for zyxin during megakaryocytic differentiation and more precisely in the regulation of the integrin mediated adhesion process in megakaryocytes. PMID- 15120621 TI - 2-O-(2-Aminoethyl)-myo-inositol 1,4,5-trisphosphate as a novel ligand for conjugation: physicochemical properties and synthesis of a new Ins(1,4,5)P(3) affinity matrix. AB - 2-O-(2-Aminoethyl)-Ins(1,4,5)P(3), (5), a novel derivative of the Ca(2+) mobilising second messenger d-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)], was synthesised from myo-inositol. 5 was found to be a potent mobiliser of intracellular Ca(2+), and an Ins(1,4,5)P(3) affinity matrix synthesised from 5 was effective at selectively binding N-terminal fragments of the Ins(1,4,5)P(3) receptor containing the intact Ins(1,4,5)P(3) binding site. The microprotonation scheme for 5 was resolved and the related constants were determined in comparison with Ins(1,4,5)P(3) and another reactive Ins(1,4,5)P(3) analogue 1-O-(2 aminoethyl-1-phospho)-Ins(4,5)P(2), (2a), by potentiometric and NMR titration methods. The (31)P and (1)H NMR titration curves for compound 5 and Ins(1,4,5)P(3) are remarkably close, indicating analogous acid-base properties and intramolecular interactions for the two compounds. The 1-phosphate-modified Ins(1,4,5)P(3) derivative 2a, on the contrary, behaves as a bisphosphorylated rather than a trisphosphorylated inositol. Thus, 5 is a new reactive Ins(1,4,5)P(3) analogue of considerable potential for investigation of the chemical biology of Ins(1,4,5)P(3)-mediated cellular signalling. PMID- 15120622 TI - Human neuroglobin interacts with flotillin-1, a lipid raft microdomain-associated protein. AB - Neuroglobin (Ngb) is a newly discovered vertebrate globin that is expressed in the brain and that can reversibly bind oxygen. It has been reported that Ngb levels increase in neurons in response to oxygen deprivation, and that it protects neurons from hypoxia. However, the mechanism of this neuroprotection remains unclear. Recently, we found that oxidized human Ngb bound to the alpha subunits of heterotrimeric G proteins (Galpha) and acted as a guanine nucleotide dissociation inhibitor for Galpha. To identify other Ngb-binding proteins, we herein screened a human brain cDNA library by using a yeast two-hybrid system. Among the plasmids isolated from positive clones, one contained an insert with 100% sequence identity to human flotillin-1. The interaction of Ngb with flotillin-1 was confirmed by glutathione S-transferase pull-down experiments. Since Galpha exists within lipid rafts critical for signal transduction and flotillin-1 recruits signaling proteins to lipid rafts, flotillin-1 might recruit Ngb to lipid rafts as a means of preventing neuronal death. PMID- 15120624 TI - Crystal structure of the glutaredoxin-like protein SH3BGRL3 at 1.6 Angstrom resolution. AB - We report the 1.6 Angstrom resolution crystal structure of SH3BGRL3, a member of a new mammalian protein family of unknown function. The observed "thioredoxin fold" of SH3BGRL3 matches the tertiary structure of glutaredoxins, even in the N terminal region where the sequence similarity between the two protein families is negligible. In particular, SH3BGRL3 displays structural modifications at the N terminal Cys-x-x-Cys loop, responsible for glutathione binding and catalysis in glutaredoxins. The loop hosts a six residue insertion, yielding an extra N terminal-capped helical turn, first observed here for the thioredoxin fold. This, together with deletion of both Cys residues, results in a substantial reshaping of the neighboring cleft, where glutathione is hosted in glutaredoxins. While not active in redox reaction and glutathione binding, SH3BGRL3 may act as an endogenous modulator of glutaredoxin activities by competing, with its fully conserved thioredoxin fold, for binding to yet unknown target proteins. PMID- 15120623 TI - Interaction of the hepatitis B virus X protein (HBx) with heat shock protein 60 enhances HBx-mediated apoptosis. AB - Understanding the function of the hepatitis B virus X protein (HBx) is fundamental to elucidating the underlying mechanisms of hepatitis and hepatocarcinogenesis caused by hepatitis B virus (HBV) infection. We identified heat shock protein 60 (Hsp60) as a novel cellular target of HBx by the combination of affinity purification and mass spectrometry. Physical interaction between HBx and Hsp60 was confirmed by standard immunoprecipitation and immunoblot methods. Analysis of HBx deletion constructs showed that amino acids 88-117 of HBx were responsible for the binding to Hsp60. Confocal laser microscopy demonstrated that HBx and Hsp60 colocalized in mitochondria. Furthermore, terminal deoxynucleotidyl transferase-mediated dUTP end labeling (TUNEL) revealed that the introduction of Hsp60 into cells facilitated HBx induced apoptosis. These findings suggest the importance of the molecular chaperon protein Hsp60 to the function of HBV viral proteins. PMID- 15120625 TI - Molecular cloning of a K(+) channel from the malaria parasite Plasmodium falciparum. AB - In most living cells, K(+) channels are important for the generation of the membrane potential and for volume regulation. The parasite Plasmodium falciparum, which causes malignant malaria, must be able to deal with large variations in the ambient K(+) concentration: it is exposed to high concentrations of K(+) when inside the erythrocyte and low concentrations when in plasma. In the recently published genome of P. falciparum, we have identified a gene, pfkch1, encoding a potential K(+) channel, which to some extent resembles the big-conductance (BK) K(+) channel. We have cloned the approximately 6000 nucleotide (nt) fragment from cDNA, studied the pattern of expression of pfkch1 throughout the intraerythrocytic part of the parasite's life-cyclus, and characterized the channel on the basis of similarity to other K(+) channels from pro- and eukaryotic organisms. This P. falciparum K(+) channel could be a potential drug target. PMID- 15120626 TI - Tbx12 regulates eye development in Xenopus embryos. AB - The regulation of vertebrate eye development requires the activity of many transcription factors. In this report, we demonstrate that the T-box factor Tbx12 is necessary for normal development of the retina. Tbx12 is expressed during early stages of retinal development in multiple species of vertebrate embryos. We injected mRNAs encoding wild type and mutant forms of Tbx12 into Xenopus embryos. The Tbx12 injected embryos exhibit multiple defects in eye development including reduced eye size and disruption of normal retinal laminar organization. Tbx12 appears to function as a repressor of transcription during eye development. Our results indicate that Tbx12 activity is required for the proper generation and organization of retinal cells in the vertebrate eye. PMID- 15120627 TI - Specific interactions of quercetin and other flavonoids with target proteins are revealed by elicited fluorescence. AB - The fluorogenic properties of quercetin and similar flavonoids common in plants were exploited to analyse their interaction with target proteins. Quercetin produced a strong fluorescent signal upon binding to bovine serum albumin (BSA) and insulin. The fluorescent signal showed saturation kinetics with increasing flavonoid concentrations indicating the presence of defined peptide binding motifs. Other tested proteins showed no fluorescence with the flavonoids. In a comparative study including 22 flavonoids the compounds with fluorogenic properties were identified using our model proteins BSA and insulin and the structural requirements for the fluorogenic property were defined. Only flavones with a high degree of hydroxylation were able to elicit fluorescence. The emitted fluorescence was strongly enhanced at alkaline pH. Finally, an attempt was made to identify intracellular target molecules in live cells. Drosophila follicles showed a distinct staining pattern thus giving evidence that high concentrations of quercetin binding proteins are present in the nuclei and are associated with the ring canals. The presented biochemical and cytological data show that the interaction of the studied flavonoids with target proteins is specific and this finding opens up new experimental possibilities to systematically identify the cellular proteins with specific binding motifs for quercetin or other fluorogenic compounds of medical interest. PMID- 15120628 TI - Glutamate modulation of human lymphocyte growth: in vitro studies. AB - Peripheral blood mononuclear cell (PBMC) proliferation induced by phytohemagglutinin, or by anti-CD3 alone or plus anti-CD28 monoclonal antibodies (mAb) was inhibited by glutamate (Glu) in a concentration-dependent manner. This inhibition was not reproduced by selective ionotropic Glu receptor agonists, whereas it was potentiated by l-buthionine-(S,R)-sulfoximine, which depletes glutathione (GSH) stores, and counteracted by 2-mercaptoethanol, a preserver of cell thiols. The inhibitory effects of Glu were related to depletion of intracellular GSH stores, since it decreased GSH levels in a concentration dependent manner. Furthermore, Glu modulated cytokine secretion by anti-CD3 mAb activated PBMC: it increased IFN-gamma (+44.3+/-8.2%) and IL-10 (+31.6+/-9.7%) secretion, whereas that of IL-2, IL-4, IL-5, and TNF-alpha was not affected. These data suggest that high levels of Glu, which can be reached in damaged tissues, modulate lymphocyte responses to activating stimuli by favouring polarization of the T helper effector response. PMID- 15120629 TI - Testosterone is a potent inhibitor of L-type Ca(2+) channels. AB - Testosterone administration is beneficial in alleviating myocardial ischaemia in men with significant coronary artery disease (CAD), a condition which is associated with hypotestosteronaemia. Infusion of physiological concentrations of testosterone into coronary arteries at angiography results in rapid vasodilatation in patients with CAD. Whilst the cardiovascular benefits of testosterone have long been documented, the underlying mechanism(s) have not yet been revealed. Here, we have investigated whether testosterone might act like widely prescribed antihypertensive dihydropyridines, as an endogenous Ca(2+) channel antagonist. To do this, we used the whole-cell patch-clamp technique to record Ca(2+) currents from the A7r5 smooth muscle cell line and HEK 293 cells stably expressing either L- or T-type Ca(2+) channels. We demonstrate that testosterone directly inhibited both native and human recombinant vascular L-type Ca(2+) channels in a manner that was voltage-independent and, crucially, displayed an IC(50) value of 38 nM, a value within the physiological range. At higher (supraphysiological) concentrations both native and human recombinant T type channels were also inhibited by testosterone. Our data indicate that testosterone acts like widely prescribed antihypertensive dihydropyridines to reduce Ca(2+) influx into vascular smooth muscle and so promote vasodilation. This effect is likely to account for its beneficial cardiovascular actions. PMID- 15120630 TI - The effect of variable domain orientation and arrangement on the antigen-binding activity of a recombinant human bispecific diabody. AB - In recent years a variety of recombinant methods have been developed for efficient production of bispecific antibodies (BsAb) in various formats. Bispecific diabody (bDAb), a 55-60 kDa molecule comprising two non-covalently associated cross-over single chain Fv (scFv) polypeptides, represents one of the most promising as well the most straightforward approaches to BsAb production. Here we constructed a bDAb, using two human scFv, 11F8 and A12, directed against the epidermal growth factor receptor (EGFR) and the insulin-like growth factor receptor (IGFR), respectively, as the building blocks. A total of 8 scFv and diabody constructs were prepared comprising the same two variable heavy (V(H)) and variable light (V(L)) chain domains but arranged in different orientations. V(H)/V(L) orientation, i.e., V(H)-linker-V(L) or V(L)-linker-V(H), showed significant effects on the expression and antigen-binding activity of scFv and monospecific diabody of both 11F8 and A12. Further, only 2 out of the 4 possible V(H)/V(L) orientations/arrangements in bDAb construction yielded active products that retain binding activity to both EGFR and IGFR. Both active bDAb preparations retained their original antigen-binding activity after incubation at 37 degrees C in mouse serum for up to 7 days, indicating excellent stability of the constructs. Taken together, our results underscore the importance of identifying/selecting optimal V(H)/V(L) orientation/arrangement for efficient production of active bDAb. PMID- 15120631 TI - Proteomic approach identifies HSP27 as an interacting partner of the hepatitis C virus NS5A protein. AB - Chronic infection by HCV is closely correlated with liver diseases such as cirrhosis, steatosis, and hepatocellular carcinoma. To understand how long-term interaction between HCV and the host leads to pathogenesis, we identified cellular proteins that interact with NS5A and NS5B using a biochemical approach. Stable cell lines that express flag-NS5A or flag-NS5B under tetracycline induction were generated. The induced flag-tagged proteins were immunoprecipitated (IP'd) and associated proteins separated on 2D gels. Protein spots that specifically co-IP'd with NS5A or NS5B were identified by mass spectrometry. HSP27 was identified as a protein that specifically co-IP'd with NS5A but not with NS5B. The N-terminal regions of NS5A (a.a. 1-181) and HSP27 (a.a. 1-122) were defined to be the domains that interact with each other. HSP27 is generally distributed in the cytoplasm. When heat shocked, HSP27 is concentrated in the ER where NS5A is co-localized. PMID- 15120632 TI - Dual actions of nitric oxide on angiogenesis: possible roles of PKC, ERK, and AP 1. AB - Regulation of angiogenesis by nitric oxide (NO) is controversial since NO has been shown to have both pro- and anti-angiogenic effects. In this study, we examined the effect of the NO donor, S-nitro-N-acetyl-penicillamine (SNAP), on in vitro angiogenesis, and the mechanisms involved: PKC activity, ERK and c-Jun phosphorylation, and AP-1 DNA binding activity, in microvascular endothelial cells. SNAP, at 0.5-4 mM, significantly and dose-dependently inhibited angiogenesis, PKC activity, and ERK and c-Jun phosphorylation up to 80%, 83%, and 63% and 73%, respectively. SNAP at concentrations > 2mM also abolished AP-1 binding activity. Lower concentrations of SNAP (0.1-0.3 mM) significantly increased angiogenesis, PKC activity, and ERK and c-Jun phosphorylation up to 46%, 60%, and 61% and 180%, respectively. These findings indicate that the dual pro- and anti-angiogenic actions of NO are dose-dependent and suggest that they are mediated by PKC and ERK acting on AP-1. PMID- 15120633 TI - Transcriptional regulation of the LAT-1/CD98 light chain. AB - LAT-1/CD98 amino acid transporter expression and activity are induced in hepatic cells deprived of arginine. The promoter dependency of this regulation was investigated. LAT-1 expression, in contrast to that of CD98 heavy chain 4F2, was actinomycin D sensitive in cells cultured without arginine. Transient transfection analysis with promoter reporter constructs including the 2 kbp LAT-1 promoter or a sub-sequence containing multiple potential amino acid response elements failed to show significant amino acid sensitivity in various cell types. Chromatin-dependency did not appear to account for this result as hepatic cell clones stably transfected with the promoter constructs showed little or no arginine or leucine responsive promoter activity. These studies suggest that while amino acid sensitivity of LAT-1 expression is transcriptionally regulated, cis elements within the proximal promoter do not directly mediate this regulation. Understanding mechanisms by which this gene responds to amino acid availability will contribute to our knowledge of how eukaryotic cells sense and respond to their environment. PMID- 15120634 TI - Novel point mutations in the mitochondrial DNA detected in patients with dilated cardiomyopathy by screening the whole mitochondrial genome. AB - Dilated cardiomyopathy (DCM) is widely accepted as a pluricausal or multifactorial disease. Because of the linkage between energy metabolism in the mitochondria and cardiac muscle contraction, it is reasonable to assume that mitochondrial abnormalities may be responsible for some forms of DCM. We analysed the whole mitochondrial genome in a series of 45 patients with DCM for alterations and compared the findings with those of 62 control subjects. A total of 458 sequence changes could be identified. These sequence changes were distributed among the whole mitochondrial DNA (mtDNA). An increased number of novel missense mutations could be detected nearly in all genes encoding for protein subunits in DCM patients. In genes coding for NADH dehydrogenase subunits the number of mtDNA mutations detected in patients with DCM was significantly increased (p < 0.05) compared with control subjects. Eight mutations were found to occur in conserved amino acids in the above species. The c.5973G > A (Ala-Trp) and the c.7042T > G (Val-Asp) mutations were located in highly conserved domains of the gene coding for cytochrome c oxidase subunit. Two tRNA mutations could be detected in the mtDNA of DCM patients alone. The T-C transition at nt 15,924 is connected with respiratory enzyme deficiency, mitochondrial myopathy, and cardiomyopathy. The c.16189T > C mutation in the D-loop region that is associated with susceptibility to DCM could be detected in 15.6% of patients as well as in 9.7% of controls. Thus, mutations altering the function of the enzyme subunits of the respiratory chain can be relevant for the pathogenesis of dilated cardiomyopathy. PMID- 15120635 TI - DNMT3B interacts with hSNF2H chromatin remodeling enzyme, HDACs 1 and 2, and components of the histone methylation system. AB - The non-random pattern of genome-wide DNA methylation in mammalian cells is established and maintained by DNA methyltransferases DNMT1, 3A, and 3B. De novo DNA methyltransferase DNMT3B is critical for embryonic development and is mutated in ICF syndrome. Despite its importance in normal cellular functioning, little is known about how DNMT3B operates in the context of chromatin. Here we demonstrate that DNMT3B associates with four chromatin-associated enzymatic activities common to transcriptionally repressed, heterochromatic regions of the genome: DNA methyltransferase, histone deacetylase, ATPase, and histone methylase activities. By immunoprecipitation and GST pull-down, we show that DNMT3B interacts with HDAC1, HDAC2, HP1 proteins, Suv39h1, and the ATP-dependent chromatin remodeling enzyme hSNF2H. Endogenous hSNF2H is also associated with DNA methyltransferase activity. These proteins co-localize extensively with DNMT3B in heterochromatic regions. Our results therefore link DNMT3B to three other components of the epigenetic machinery and provide important insights into how DNA methylation patterns may be established within the chromatin environment. PMID- 15120636 TI - Potent inhibition of human cardiac potassium (HERG) channels by the anti-estrogen agent clomiphene-without QT interval prolongation. AB - The acquired form of the long-QT syndrome (LQTS) is a major safety consideration for the development and subsequent use of both cardiac and non-cardiac drugs; it is usually associated with pharmacological inhibition of cardiac HERG-encoded potassium channels. Clomiphene is an anti-estrogen agent used extensively in the treatment of infertility and is not associated with a risk of QT interval prolongation, in contrast to a structurally related compound tamoxifen. We describe here a potent inhibitory effect (IC(50) = 0.18 microM) of clomiphene on HERG ionic current (I(HERG)) recorded from a mammalian cell line expressing HERG channels. Inhibition of I(HERG) by clomiphene showed voltage-dependence and developed quickly following membrane depolarisation, indicating contingency of block on HERG channel gating. At 100 nM, clomiphene and the related anti-estrogen tamoxifen produced similar levels of I(HERG) blockade (p > 0.05). Experiments on guinea-pig isolated perfused hearts revealed that, despite its inhibitory action on I(HERG), clomiphene produced no significant effect at 1 microM on uncorrected QT interval (p > 0.1) nor on rate-corrected QT interval (QT(c); p > 0.1 for QT(c) determined using Van de Water's formula). The disparity between clomiphene's potent I(HERG) inhibition and its lack of effect on the QT interval underscores the notion that I(HERG) pharmacology may best be used alongside other screening methods when investigating the QT-prolonging tendency and related cardiotoxicity of non-cardiac drugs. PMID- 15120637 TI - Incorporation of biodegradable nanoparticles into human airway epithelium cells in vitro study of the suitability as a vehicle for drug or gene delivery in pulmonary diseases. AB - PURPOSE: Nanoparticles are able to enhance drug or DNA stability for purposes of optimised deposition to targeted tissues. Surface modifications can mediate drug targeting. The suitability of nanoparticles synthesised out of porcine gelatin, human serum albumin, and polyalkylcyanoacrylate as drug and gene carriers for pulmonary application was investigated in vitro on primary airway epithelium cells and the cell line 16HBE14o-. METHODS: The uptake of nanoparticles into these cells was examined by confocal laser scan microscopy (CLSM) and flow cytometry (FACS). Further the cytotoxicity of nanoparticles was evaluated by an LDH-release-test and the inflammatory potential of the nanoparticles was assessed by measuring IL-8 release. RESULTS: CLSM and FACS experiments showed that the nanoparticles were incorporated into bronchial epithelial cells provoking little or no cytotoxicity and no inflammation as measured by IL-8 release. CONCLUSIONS: Based on their low cytotoxicity and the missing inflammatory potential in combination with an efficient uptake in human bronchial epithelial cells, protein based nanoparticles are suitable drug and gene carriers for pulmonary application. PMID- 15120639 TI - Modulation of actomyosin contractility by myosin light chain phosphorylation/dephosphorylation through Rho GTPases signaling specifies axon formation in neurons. AB - Actin depolymerization through Rho GTPases or exogenous mechanical tension has been suggested as a key determinant for the first step of neuronal polarization, the axonogenesis, in which one of the neurites starts to grow becoming the axon. The underlying mechanism and the relationship between two forces in the cells, however, are mostly unknown. Here, we report that the myosin-dependent contractility is a common effector between two forces and a critical determinant in axonogenesis and neuronal polarization. We have found that inhibition of myosin ATPase activity and modulation of myosin light chain phosphorylation/dephosphorylation through Rho GTPases signaling induced multiple axons. Moreover, overexpression of wild-type myosin light chain kinase dramatically increased filopodial structures and produced multi-axonal structures. Our results suggest that MLC phosphorylation/dephosphorylation through Rho GTPases signaling modulates the actomyosin contractility, and then in turn provides a physiological tension in neurons to induce axon. PMID- 15120638 TI - Aryl hydrocarbon receptor-mediated induction of microsomal drug-metabolizing enzyme activity by indirubin and indigo. AB - Indirubin and indigo, which are thought to be natural ligands for aryl hydrocarbon receptor (AhR), showed marked AhR ligand activities in a reporter gene assay using recombinant yeast. Their activities were comparable with or more potent than that of 2,3,7,8-tetrachlorodibenzo-p-dioxin. When indirubin and indigo were administered to mice, ethoxyresorufin-O-dealkylase and methoxyresorufin-O-dealkylase activities in the liver were increased, but subsequently decreased within 2 days. Indirubin was more potent than indigo. Levels of cytochrome P450 1A1/2 proteins and mRNAs in the liver of mice dosed with indirubin were also enhanced. These enhancing effects of indirubin and indigo were not observed in AhR knock-out mice. Ethoxyresorufin-O-dealkylase and methoxyresorufin-O-dealkylase activities in rat hepatocytes and HepG2 cells were enhanced by the addition of indirubin or indigo, but less potently than by 2,3,7,8-tetrachlorodibenzo-p-dioxin. Indigocarmine, a sulfate derivative of indigo, which is used as food additive, did not show these inducing effects on drug-metabolizing enzymes. Our results suggest that indirubin and indigo act as inducers for cytochrome P450 1A1/2 mediated by AhR in mammals in vivo. PMID- 15120640 TI - Analysis of recombinant human saposin A expressed by Pichia pastoris. AB - Saposins (SAPs) are small glycoproteins required for activation of sphingolipid hydrolysis by lysosomal enzymes. Four SAPs, SAP-A, -B, -C, and -D, are proteolytically cleaved from a single gene product termed prosaposin. The mature coding sequence of human SAP-A tagged with 6-histidine was expressed in Pichia pastoris and the recombinant protein was purified from the culture supernatant by simple purification steps with an immobilized metal ion affinity column, a Concanavalin A column, and reversed-phase HPLC. Secreted SAP-A contained both glycosylated and nonglycosylated forms. Both forms of SAP-A activated galactocerebroside and 4-methylumbelliferyl beta-d-glucoside hydrolysis by galactocerebrosidase and glucocerebrosidase. SAP-A expressed in P. pastoris should be useful for further structural and functional analysis of this protein. PMID- 15120641 TI - Fibronectin-induced COX-2 mediates MMP-2 expression and invasiveness of rhabdomyosarcoma. AB - Although accumulating evidence suggests the importance of cyclooxygenase-2 (COX 2) and prostaglandin E(2) (PGE(2)) in the pathogenesis of many cancers, the mechanism by which this enzyme and its metabolite promote cancer progression is unknown. In this study, we investigated the role of COX-2 in fibronectin-induced up-regulation of rhabdomyosarcoma matrix metalloproteinase (MMP)-2 activity and cellular invasiveness. We tested three human rhabdomyosarcoma cell lines: RMS559, RD, and SJRH30. Cell attachment to fibronectin up-regulated both COX-2 expression and PGE(2) production and concomitantly enhanced MMP-2 activity. Exogenous PGE(2) stimulated MMP-2 promoter activity, increased MMP-2 expression, and increased cellular invasiveness. Aspirin and rofecoxib (non-selective and selective COX-2 inhibitor, respectively) each abolished fibronectin-associated induction of MMP-2 and induced dose-dependent reductions in cellular invasiveness. These data implicated a role for inducible COX-2 and PGE(2) in the regulation of rhabdomyosarcoma cellular invasiveness and MMP-2 activity. PMID- 15120642 TI - Analyzing and enhancing mRNA translational efficiency in an Escherichia coli in vitro expression system. AB - The dependence of efficiency of translation initiation on mRNA sequence parameters was investigated in an Escherichia coli in vitro expression system. We designed a large-scale expression experiment focussing on the influence of sequence variations in the translated region (TR) of the mRNA without changing the 5'-untranslated region (5'-UTR). The level of translated protein from 756 expression constructs was measured and the influence of a large number of possible effector attributes was statistically analyzed. Base exchanges immediately adjacent to the start codon up to nucleotide (+)25 had a profound effect on translational efficiency. Correlation analysis revealed a significant dependence on base pair probability and G+C content on the expression level, indicating that mRNA secondary structure in this region hampers translation. Using our training data, we developed a methodology to predict and improve the translation efficiency of open reading frames (ORFs). PMID- 15120643 TI - YY1 binding to a subset of p53 DNA-target sites regulates p53-dependent transcription. AB - The tumor suppressor protein p53 regulates gene transcription through binding to specific DNA-target sites. We here demonstrate that a subset of these sites is targeted by another DNA-binding factor. Binding specificity, reactivity with specific antibodies, and experiments with purified protein identified the factor as the multifunctional transcription regulator YY1. The YY1 core binding sequence ACAT is present in the center of p53-half-binding sites in the p21 and GADD45 genes regulating growth arrest and DNA repair, respectively, but is absent in those of the Bax gene critical for apoptosis. In transfection experiments YY1 inhibits p53-activated transcription from the p53-binding site that contains the ACAT sequence. YY1 and p53 are colocalized around the nucleoli and in discrete nuclear domains in PC12 cells undergoing apoptosis. YY1 might attenuate p53 dependent transcription from a subset of p53-target genes and this may be relevant for directing cells either to growth arrest or apoptosis upon p53 activation. PMID- 15120644 TI - Expression patterns of IL-10 ligand and receptor gene families provide leads for biological characterization. AB - The expression patterns of the IL-10 ligand and receptor genes were examined in normal and transformed cell lines of human hematopoietic and non-hematopoietic origin. IL-10 family ligands, IL-10, IL-19, IL-20, IL-22, IL-24 and IL-26 were predominantly expressed by hematopoietic cells. IL-10, IL-24 and IL-26 were produced by both monocytes and T cells, IL-19 and IL-20 were produced by monocytes whereas IL-22 was produced mainly by activated T cells. The receptors of the IL-10 family, IL-10R1, IL-10R2, IL-20R1, IL-20R2, IL-22R1 and IL-22 BP were also expressed in a distinct pattern when probed on these cell lines. The expression of IL-10R2 was ubiquitous whereas IL-10R1 was predominantly expressed on hematopoietic cells, including, T cells, B cells, NK cells, monocytes and dendritic cells. IL-20R1, IL-20R2 and IL-22R1 were absent or expressed at extremely low levels on cells of the hematopoietic lineage. These receptors were mainly found on epithelial and stromal cells fibroblasts of various tissues. Interestingly, IL-22BP was quite specifically expressed by dendritic cells. These data point to a function of the novel IL-10 family members in communication and interaction between cells of the hematopoietic and non-hematopoietic lineages, a role quite distinct from the immunomodulating effects of IL-10 itself. PMID- 15120645 TI - Full house: 12 receptors for 27 cytokines. AB - With the sequencing of the human genome nearing completion, it appears that all members of the class II cytokine receptor family (CRF2) have been identified and partially characterized. The entire family is composed of exactly one dozen members. Eleven of them combine as various heterodimers to transduce signals across the cellular membrane for 27 cytokines divided into four structurally related groups: 6 cytokines of the IL-10 family, 17 type I IFNs, 1 type II IFN and 3 IFN-lambdas. The last CRF2 member is the soluble receptor which can neutralize the action of one of the cytokines of the IL-10 family, IL-22. Although the extracellular domains of all CRF2 proteins reveal primary and structural homology, their intracellular domains are very dissimilar. Nevertheless, signaling events induced through various combinations of CRF2 subunits partially overlap, leading to the induction of overlapping but cytokine specific biological activities. PMID- 15120646 TI - Interleukin-26. AB - Interleukin-26 (IL-26), initially termed AK155, is a cellular sequence homolog to IL-10 belonging to the IL-10 cytokine family. Together with the related genes for interferon-gamma and IL-22/IL-TIF, il-26 maps to the human chromosomal region 12q15. The il-26 gene is one of the few differentially expressed genes specifying human T cells after growth-transformation with herpesvirus saimiri, a tumor virus of neo-tropical squirrel monkeys. Only herpesvirus saimiri-transformed T cells have been found to strongly over-express il-26 and to release the protein into the tissue culture supernatant. In a series of other T-cell lines and in native peripheral blood cells, il-26 is transcribed at low levels, but it is not detectable in B cells. Similarly to IL-10, the IL-26 protein forms homo-dimers. IL-26 is a candidate to contribute to the transformed phenotype of human T cells after infection by herpesvirus saimiri. Moreover, the T-lymphokine IL-26 is highly likely to play a role in normal and pathological hematology or immunology. PMID- 15120647 TI - Human interleukin-19 and its receptor: a potential role in the induction of Th2 responses. AB - Interleukin-19 (IL-19) is a newly discovered member of the IL-10 family of ligands whose function is presently undefined. We recently described its cloning and initial characterization and in so doing, noted that the induction of IL-19 by LPS in human monocytes was down-regulated by interferon-gamma (IFN-gamma) and up-regulated by IL-4. This preliminary observation led us to speculate that IL-19 may play a role in the Th1/Th2 system and we examined this hypothesis further. Our results suggested that IL-19 is able to influence the maturation of human T cells. CD4+ T-cells resulting from SEB stimulation in the presence of IL-19 contained a higher proportion of IL-4 producing cells than those developing in the absence of IL-19. This observation was complimented by the observation that fewer IFN-gamma cells accrued in the presence of IL-19, thereby suggesting that IL-19 altered the balance of Th1/Th2 cells in favour of Th2. Furthermore, in whole PBMC cultures, IL-19 up-regulated IL-4 and down-regulated IFNgamma in a dose-dependent manner. These results are presented here in review format, in the context of an overall discussion of IL-19 and its receptor. PMID- 15120648 TI - Interleukin-20. PMID- 15120649 TI - Cytokine and tumor cell apoptosis inducing activity of mda-7/IL-24. AB - Melanoma Differentiation Associated gene-7 (mda-7)/IL-24 has shown potent tumor cell apoptosis inducing capacity in multiple cancers, making it a strong candidate for use as a human cancer gene therapeutic. Several independent studies have currently documented and confirmed mda-7/IL-24's cytokine nature including presence of a canonical secretory signal peptide, processing and secretion of the molecule by cells and it's binding to specific interleukin receptors on the cell surface. Receptor binding has been shown to activate the JAK/STAT signal transduction pathway with concomitant stimulation of STAT 1 and 3 transactivators. The physiological role(s) of this molecule in modulating immune responses, as a member of the IL-10 family of cytokines, is not well documented and most current information pertains to its apparently restricted expression patterns in specific cell types with immunomodulatory activity. On the other hand, several additional signal transduction pathways were modulated when cells overexpress mda-7/IL-24, not all of which are necessarily downstream of mda-7/IL 24 induced JAK/STAT activation. A summary of the current status of information is presented to provide a perspective for the cytokine-related properties of mda 7/IL-24 in correlation to its tumor cell apoptosis inducing activity. Moreover, new evidence has surfaced pointing toward apoptosis induction via mechanisms independent of cytokine activity-related JAK/STAT activation. PMID- 15120650 TI - MDA-7/IL-24 is a unique cytokine--tumor suppressor in the IL-10 family. AB - The melanoma differentiation associated gene-7 (mda-7) cDNA was isolated by virtue of being induced during melanoma differentiation. Initial gene transfer studies convincingly demonstrated potent antitumor effects of mda-7. Further studies showed that the mechanism of antitumor activity was due to induction of apoptosis. Most striking was the tumor-selective killing by mda-7 gene transfer- normal cells were unaffected by Adenoviral delivery of mda-7 (Ad-mda7). A variety of molecules implicated in apoptosis and intracellular signaling are regulated by Ad-mda7 transduction. Different apoptosis effector proteins are regulated in different tumor types, suggesting that Ad-mda7 may regulate various signaling pathways. mda-7 encodes a secreted protein, MDA-7, which has now been designated as IL-24, and is a novel member of the IL-10 cytokine family. MDA-7/IL-24 protein is actively secreted from cells after mda-7 gene transfer. In human peripheral blood mononuclear cells (PBMC), STAT3 activation by MDA-7/IL-24 is followed by elaboration of secondary Th1 cytokines, demonstrating that MDA-7/IL-24 is a pro Th1 cytokine. Furthermore, MDA-7/IL-24 is antagonized by the prototypic Th2 cytokine IL-10. MDA-7/IL-24 protein is endogenously expressed in cultured NK and B-cells and is also expressed in dendritic cells in tissues. MDA-7/IL-24 protein is expressed in nevi and melanoma primary tumors, to varying degrees, but is rarely expressed in malignant melanoma or other human tumors evaluated. Indeed, loss of MDA-7/IL-24 protein expression correlates strongly with melanoma tumor invasion and disease progression. The "bystander" effects proposed for MDA-7/IL 24 protein include immune stimulation, antiangiogenesis and receptor-mediated cytotoxicity. Thus, mda-7 is a unique multifunctional cytokine in the IL-10 family and may have potent antitumor utility in a clinical setting. PMID- 15120651 TI - IL-22, a Th1 cytokine that targets the pancreas and select other peripheral tissues. AB - IL-22, also termed IL-TIF, is a member of the IL-10 family of cytokines. Its principal source appears to be memory CD4 T cells with a Th1 polarized phenotype. IL-22 induces its signals through a two-component receptor comprised of IL-22R1 and CRF2-4/IL10Rb. Both of these receptor components also participate in separate receptor complexes specific for other IL-10 family cytokines. Because CRF2-4 exhibits ubiquitous expression, the tropism of IL-22 action appears to be dictated by the expression of IL-22R1. IL-22R1 has a highly restricted expression pattern. Its highest expression, by far, is in the acinar cell population of the pancreas. Lower, but still functional, levels of expression are also observed in skin, colon, liver, and kidney. The responses that have been observed to date for IL-22 resemble the "acute phase" type responses elicited by IL-6, suggesting that IL-22 might be appropriately considered as a T cell-derived IL-6-like activity having distinct target cell specificity. The functional role of this system remains unclear, but it is likely that the responses elicited by this cytokine serve to contribute both to acute host defense against pathogens and to safeguard vulnerable target tissues under conditions of stress. PMID- 15120652 TI - Interleukin-22 activates STAT3 and induces IL-10 by colon epithelial cells. AB - Human interleukin-22 (IL-22), a cytokine with structural homology to IL-10, is produced by activated T cells. The IL-22 receptor complex consists of a ligand binding chain, the IL-22R1 and a signal-transducing chain, the IL-10R2. The aim of this study is to identify potential target cells and associated biological activity of IL-22 by identifying cell types that specifically express high levels of IL-22R1 as the expression of IL-10R2 is ubiquitous. Expression of IL-22R1 mRNA, as analyzed by real time quantitative polymerase chain reaction (PCR), was observed in human tumor cell lines of stromal or epithelial origin derived from liver, pancreas, colon and lung tissue. Furthermore, we examined the ability of IL-22 to activate the JAK-Signal Transducer and Activator of Transcription (STAT) pathway in epithelial cells of the colon. IL-22 induced the phosphorylation of STAT1 and STAT3 in Colo205, a colon epithelial cell line. Consequently, IL-22 upregulated mRNA for Suppressor of Cytokine Signaling 3 (SOCS3), a STAT3 responsive gene. Further analyses, by real time quantitative PCR, on a panel of chemokines and immune function related genes revealed that IL-22 induced expression of the acute phase proteins alpha-Antichymotrypsin and Serum Amyloid A, as well as IL-10 mRNA and protein production by Colo205. Induction of IL-10 by IL-22, in Colo205 cells, could be inhibited in the presence of a neutralizing antibody against IL-10R2. IL-22-mediated effects on the Colo205 cells were also inhibited in the presence of IL-22 binding protein (IL-22BP), a soluble receptor with structural similarity to IL-22R1. The high levels of expression of IL-22R1 observed in epithelial cells of the colon and the ability of IL-22 to upregulate production of acute phase proteins and IL-10 in Colo205 cells, suggest a functional role for IL-22 in intestinal inflammation. PMID- 15120653 TI - Temporal associations between interleukin 22 and the extracellular domains of IL 22R and IL-10R2. AB - Interleukin 22 (IL-22) is a cytokine induced during both innate and adaptive immune responses. It can effect an acute phase response, implicating a role for IL-22 in mechanisms of inflammation. IL-22 requires the presence of the IL-22 receptor (IL-22R) and IL-10 receptor 2 (IL-10R2) chains, two members of the class II cytokine receptor family (CRF2), to effect signal transduction within a cell. We studied the interaction between human IL-22 and the extracellular domains (ECD) of its receptor chains in an enzyme-linked immunoabsorbant assay (ELISA) based format, using biotinylated IL-22 (bio-IL-22) and receptor-fusions containing the ECD of a receptor fused to the Fc of hIgG1 (IL-22R-Fc and IL-10R2 Fc). IL-22 has measurable affinity for IL-22R-Fc homodimer and undetectable affinity for IL-10R2. IL-22 has substantially greater affinity for IL-22R/IL-10R2 Fc heterodimers. Further analyses involving sequential additions of receptor homodimers and cytokine indicates that the IL-10R2(ECD) binds to a surface created by the interaction between IL-22 and the IL-22R(ECD), and thereby further stabilizes the association of IL-22 within this cytokine-receptor-Fc complex. Both a neutralizing rat monoclonal antibody, specific for human IL-22, and human IL-22BP-Fc, an Fc-fusion of the secreted IL-22 binding-protein and proposed natural antagonist for IL-22, bind to similar cytokine epitopes that may overlap the binding site for IL-22R(ECD). Another rat monoclonal antibody, specific for IL-22, binds to an epitope that may overlap a separate binding site for IL 10R2(ECD). We propose, based on this data, a temporal model for the development of a functional IL-22 cytokine-receptor complex. PMID- 15120654 TI - Towards the development of a conceptual distance metric for the UMLS. AB - The objective of this work is to investigate the feasibility of conceptual similarity metrics in the framework of the Unified Medical Language System (UMLS). We have investigated an approach based on the minimum number of parent links between concepts, and evaluated its performance relative to human expert estimates on three sets of concepts for three terminologies within the UMLS (i.e., MeSH, ICD9CM, and SNOMED). The resulting quantitative metric enables computer-based applications that use decision thresholds and approximate matching criteria. The proposed conceptual matching supports problem solving and inferencing (using high-level, generic concepts) based on readily available data (typically represented as low-level, specific concepts). Through the identification of semantically similar concepts, conceptual matching also enables reasoning in the absence of exact, or even approximate, lexical matching. Finally, conceptual matching is relevant for terminology development and maintenance, machine learning research, decision support system development, and data mining research in biomedical informatics and other fields. PMID- 15120655 TI - Cognitive schema and naturalistic decision making in evidence-based practices. AB - A recent article in this journal proposed a naturalistic approach to decision making that overcomes problems intrinsic to classical decision theory. The approach emphasizes cognitive and multi-level processes, the development of expert reasoning, and the role of decision support in individual and organizational decision making. The current paper builds on this effort by suggesting a naturalistic, multi-level, theory that can facilitate the dissemination of evidence-based practices (EBPs). The paper presents "Image Theory," a theory that has been extensively investigated in other disciplines, but has yet to be utilized in medical decision research. It is suggested that its rich, empirically tested, distinctions among kinds of cognitive and organizational processes and types of decisions and tasks make Image Theory especially valuable in describing impediments to implementing EBPs. The paper discusses how naturalistic theory can assist clinicians, administrators, researchers, and policy makers in achieving a balance between evidence-based medicine and patient-centered practice. PMID- 15120656 TI - A computational TW3 classifier for skeletal maturity assessment. A Computing with Words approach. AB - This paper proposes a fuzzy methodology to translate the natural language descriptions of the TW3 method for bone age assessment into an automatic classifier. The classifier is built upon a modified version of a fuzzy ID3 decision tree. No large data records are needed to train the classifier, i.e., to find out the classification rules, since the classifier is built upon rules given by the TW3 method. Only small data records are needed to fine-tune the fuzzy sets used to implement the rulebase. PMID- 15120657 TI - An ontology of randomized controlled trials for evidence-based practice: content specification and evaluation using the competency decomposition method. AB - Randomized controlled trials (RCTs) are one of the least biased sources of clinical research evidence, and are therefore a critical resource for the practice of evidence-based medicine. With over 10,000 new RCTs indexed in Medline each year, knowledge systems are needed to help clinicians translate evidence into practice. Common ontologies for RCTs and other domains would facilitate the development of these knowledge systems. However, no standard method exists for developing domain ontologies. In this paper, we describe a new systematic approach to specifying and evaluating the conceptual content of ontologies. In this method, called competency decomposition, the target task for an ontology is hierarchically decomposed into subtasks and methods, and the ontology content is specified by identifying the domain information required to complete each of the subtasks. We illustrate the use of this competency decomposition approach for the content specification and evaluation of an RCT ontology for evidence-based practice. PMID- 15120658 TI - Fever detection from free-text clinical records for biosurveillance. AB - Automatic detection of cases of febrile illness may have potential for early detection of outbreaks of infectious disease either by identification of anomalous numbers of febrile illness or in concert with other information in diagnosing specific syndromes, such as febrile respiratory syndrome. At most institutions, febrile information is contained only in free-text clinical records. We compared the sensitivity and specificity of three fever detection algorithms for detecting fever from free-text. Keyword CC and CoCo classified patients based on triage chief complaints; Keyword HP classified patients based on dictated emergency department reports. Keyword HP was the most sensitive (sensitivity 0.98, specificity 0.89), and Keyword CC was the most specific (sensitivity 0.61, specificity 1.0). Because chief complaints are available sooner than emergency department reports, we suggest a combined application that classifies patients based on their chief complaint followed by classification based on their emergency department report, once the report becomes available. PMID- 15120660 TI - A critical problem. PMID- 15120659 TI - Incorporating ideas from computer-supported cooperative work. AB - Many information systems have failed when deployed into complex health-care settings. We believe that one cause of these failures is the difficulty in systematically accounting for the collaborative and exception-filled nature of medical work. In this methodological review paper, we highlight research from the field of computer-supported cooperative work (CSCW) that could help biomedical informaticists recognize and design around the kinds of challenges that lead to unanticipated breakdowns and eventual abandonment of their systems. The field of CSCW studies how people collaborate with each other and the role that technology plays in this collaboration for a wide variety of organizational settings. Thus, biomedical informaticists could benefit from the lessons learned by CSCW researchers. In this paper, we provide a focused review of CSCW methods and ideas we review aspects of the field that could be applied to improve the design and deployment of medical information systems. To make our discussion concrete, we use electronic medical record systems as an example medical information system, and present three specific principles from CSCW: accounting for incentive structures, understanding workflow, and incorporating awareness. PMID- 15120661 TI - Preservation of fertility in young women treated for cancer. PMID- 15120662 TI - Do systemic cancer treatments affect cognitive function? PMID- 15120663 TI - Do beta-tubulin pseudogenes really matter? PMID- 15120664 TI - What is the effect of systemic anticancer treatment on cognitive function? AB - Treatment regimens for solid tumours have been extensively investigated for their physical toxic effects, but far less is known about the potential impairment of cognitive function by anticancer treatment regimens. Here, we review published studies that examined cognitive function in adult patients receiving systemic therapy for solid tumours. Our review suggests that patients can experience cognitive changes related to their treatment. However, several studies had methodological limitations, such as use of a limited sample size, lack of baseline assessment, and lack of control for potential confounding factors. Better designed clinical trials are required so that the difficulties patients face in terms of reduced cognitive function as a result of anticancer treatment can be fully elucidated. These trials should have sufficient statistical power and, importantly, should also be prospective. PMID- 15120665 TI - Use of chemotherapy during human pregnancy. AB - When cancer is diagnosed in a pregnant woman, life-saving chemotherapy for the mother poses life-threatening concerns for the developing fetus. Depending on the type of cancer and the stage at diagnosis, chemotherapy cannot necessarily be delayed until after delivery. Women diagnosed with acute lymphoblastic leukaemia who decline both termination and chemotherapy often die with the previable fetus in utero. Safe use of chemotherapy, especially during the second and third trimester, have been reported, and pregnant women with cancer can accept therapy without definite neonatal harm. Here, we review the use of chemotherapy in pregnancy by trimester of exposure and summarise neonatal outcomes, including malformations, perinatal complications, and oldest age of neonatal follow-up. We will also discuss the modes of action of the drugs used and look at the multiagent regimens recommended for use during pregnancy. PMID- 15120666 TI - Monoclonal antibodies as therapeutic agents for cancer. AB - Targeted therapies are the focus of much research in oncology. After the development of imatinib for the treatment of chronic myeloid leukaemia, biological therapies that target tumour-associated antigens give hope for improvement of survival in many cancers. At the American Society of Clinical Oncology (ASCO) conference in 2003, data for the antibodies bevacizumab and cetuximab highlighted promising results in clinical trials, including an improvement in survival for metastatic colorectal cancer. Positive results for other antibodies in various stages of clinical development provide hope that anticancer antibodies will have an effect on clinical oncology practice in the next 10 years. PMID- 15120667 TI - Environmental, genetic, and molecular features of prostate cancer. AB - Prostate cancer is the sixth most common cancer in the world and the third leading cause of cancer in men. The increase in the understanding of prostate carcinogenesis over the past 15 years has helped to define crucial steps in the natural history of the disease, namely initiation and progression to androgen independence. This heterogeneous disease encompasses a range of environmental and familial factors, which provides strong support for the use of chemopreventive strategies. Most patients with advanced prostate cancer are treated with androgen deprivation therapy, which leads to a striking regression of androgen-responsive cancer cells. A transition from an androgen-responsive to an androgen unresponsive stage is seen during the clinical course in almost all patients with prostate cancer. This transition also signals a substantial worsening of prognosis. Here, we review the most important findings in prostate carcinogenesis and the molecular anomalies associated with the androgen-refractory stage. PMID- 15120668 TI - Clinical germline genetic testing for melanoma. AB - Clinical genetic testing for mutations in CDKN2A (cyclin-dependent kinase inhibitor 2A), a melanoma susceptibility gene, is now available. The International Melanoma Genetics Consortium advocates that genetic testing for CDKN2A should be done only as part of a research protocol. Experience with genetic testing for other cancer-susceptibility genes indicates that CDKN2A testing has enormous potential for the prevention and detection of a deadly disease. However, clinicians need to understand the benefits and shortcomings of clinical CDKN2A testing so that it can be used advantageously. Here, we examine whether CDKN2A meets the recommendations of the American Society of Clinical Oncology (ASCO) for cancer-susceptibility genetic testing. Although genetic testing for hereditary melanoma should, whenever possible, occur within research protocols, it might be successfully done outside of research protocols if attention is paid to selection, education, and counselling needs of patients; valid test interpretation; and the changing of medical management in appropriate individuals. PMID- 15120669 TI - Dexrazoxane for anthracycline extravasation and GM-CSF for skin ulceration and wound healing. PMID- 15120670 TI - Irinotecan-induced interstitial pneumonia. PMID- 15120671 TI - Ocular involvement in neuroblastoma: not always metastasis. PMID- 15120672 TI - The future of institutional review boards. PMID- 15120673 TI - The art of the insult. PMID- 15120674 TI - To sleep, perchance to gain creative insight? AB - The development of mathematical insight, the knack for discovering novel solutions to mathematical problems, might be one of the most erudite forms of learning that we can hope to achieve. However, Wagner and his colleague now report that a night of sleep after being exposed to a class of mathematical problems more than doubles the likelihood of discovering just such a novel solution. PMID- 15120675 TI - Something funny happened to reward. AB - The human reward system has been shown to be activated by a wide range of reinforcers, including food, money, sex, drugs, and beauty. Now, a recent fMRI study has found mesolimbic reward activation associated with humorous cartoons, providing a neurobiological link between theories of humour and hedonic processes in the brain. PMID- 15120676 TI - Sociality and the evolution of intelligence. AB - Two recently published studies provide important new data relevant to the evolution of human intelligence. Both studies of social behavior in baboons, Bergman et al. demonstrated that baboons use two criteria simultaneously to classify other troop members, and Silk et al. showed that highly social female baboons have higher reproductive success than less social females. Taken together, these studies provide strong evidence for the importance of social context in cognitive evolution. PMID- 15120677 TI - Remembrance of futures past. AB - Much behavioural and physiological evidence suggests that the hippocampus encodes space. Puzzlingly, however, hippocampal damage also disrupts episodic memory. A recent study shows how these two faculties might be related, finding that the spatial firing of hippocampal 'place cells' is sometimes modulated by what the animal has recently done or what it will do next. Thus, the cells encode something resembling a context, or episode, collectively forming a potential substrate for episodic memory. PMID- 15120678 TI - Mining event-related brain dynamics. AB - This article provides a new, more comprehensive view of event-related brain dynamics founded on an information-based approach to modeling electroencephalographic (EEG) dynamics. Most EEG research focuses either on peaks 'evoked' in average event-related potentials (ERPs) or on changes 'induced' in the EEG power spectrum by experimental events. Although these measures are nearly complementary, they do not fully model the event-related dynamics in the data, and cannot isolate the signals of the contributing cortical areas. We propose that many ERPs and other EEG features are better viewed as time/frequency perturbations of underlying field potential processes. The new approach combines independent component analysis (ICA), time/frequency analysis, and trial-by-trial visualization that measures EEG source dynamics without requiring an explicit head model. PMID- 15120679 TI - The universal psychology of kinship: evidence from language. AB - Kinship is central to social organization in many societies; how people think about kinship should be relevant to social cognition generally. One window onto the mental representation of kinship is afforded by variation and universals in terms for kin. Kin terminologies are commonly organized around binary distinctive features, and terms for some types of kin are consistently linguistically marked. These observations can be formalized in the newly developed framework of linguistic Optimality Theory: permutations in the rank order of a small set of constraints generate basic types of kin terminology without over-generating rare or non-existent types. The result, I argue, is evidence for an innate faculty of social cognition (including several universal schemas of social relationships), apparently shaped by several kinds of genetic kin selection. PMID- 15120680 TI - Social cognitive neuroscience: where are we heading? AB - Humans crave the company of others and suffer profoundly if temporarily isolated from society. Much of the brain must have evolved to deal with social communication and we are increasingly learning more about the neurophysiological basis of social cognition. Here, we explore some of the reasons why social cognitive neuroscience is captivating the interest of many researchers. We focus on its future, and what we believe are priority areas for further research. PMID- 15120681 TI - The brain circuitry of attention. AB - From an operational perspective, attention is a matter of organizing multiple brain centres to act in concert on the task at hand. Taking focal visual attention as an example, recent anatomical findings suggest that the pulvinar might act as a remote hub for coordinating spatial activity within multiple cortical visual maps. The pulvinar can, in turn, be influenced by signals originating in the frontal and parietal eye fields, using common visuomotor neural circuitry, with the superior colliculus acting as an important link. By identifying a complex, real neural architecture ('RNA') model for attention, it is possible to integrate several different modes of operation - such as parallel or serial, bottom-up or top-down, preattentive or attentive - that characterize conflicting cognitive models of attention in visual search paradigms. PMID- 15120682 TI - Disfluencies and human language comprehension. AB - Spoken language contains disfluencies, which include editing terms such as uh and um as well as repeats and corrections. In less than ten years the question of how disfluencies are handled by the human sentence comprehension system has gone from virtually ignored to a topic of major interest in computational linguistics and psycholinguistics. We discuss relevant empirical findings and describe a computational model that captures how disfluencies influence parsing and comprehension. The research reviewed shows that the parser, which presumably evolved to handle conversations, deals with disfluencies in a way that is efficient and linguistically principled. The success of this research program reinforces the current trend in cognitive science to view cognitive mechanisms as adaptations to real-world constraints and challenges. PMID- 15120683 TI - Long-term follow-up, neurological outcome and survival rate in 28 Nordic patients with glutaric aciduria type 1. AB - All 28 patients, 13 females and 15 males, with glutaric aciduria type 1 diagnosed between 1975 and 2001 in Denmark, Finland, Norway and Sweden were identified and studied retrospectively until 2001. Mass screening was not performed. Three were sibling cases. Prenatal enzymatic diagnosis performed in 11 pregnancies led to termination in one. The median follow-up time was 14 years. Six patients had died. At 10 years of age the cumulative survival rate was 89% and at 35 years 44%. The dominating neurological sign was dystonia in 20 and dyskinesia in 4. Three had only slight spastic signs and information was missing in one. The head circumference at birth was significantly larger than normal and increased significantly until 6 months of age. The onset was acute encephalopathic in 24 patients and insidious in 3. From the time of diagnosis, all patients but one were prescribed protein restriction and/or a diet low in lysine and tryptophan. Riboflavine and/or carnitine supplementation were given to 25. Neurological deficits did not improve on the offered treatment. Deterioration may have been averted by intense acute metabolic treatment in a few patients. Dystonia correlated significantly to absence of speech but not to cognitive function. Severe disability, including motor, cognitive and speech functions, correlated significantly with acute onset, dystonia and mortality, and weakly with a deteriorating course, but not with age at onset, diagnosis, or follow-up, nor to head size. Results from future population studies derived from mass screening will have to relate to clinical diagnostic series of the kind presented here. PMID- 15120684 TI - Intermittent oral diazepam prophylaxis in febrile convulsions: its effectiveness for febrile seizure recurrence. AB - In order to evaluate the effectiveness of diazepam for the reduction in the recurrence of febrile seizures we carried out a prospective study in two groups of children; Group A: 45 children (25 female, 20 male), receiving oral prophylaxis with diazepam, and Group B: 65 children (35 female, 30 male) who did not receive any oral prophylaxis. All subjects of both groups were followed for at least 4 years and finally re-evaluated at the mean age of 6.7+/-1.4 years. Among the patients of Group A, recurrent febrile seizures (FS) occurred in five of the 45 children (11.1%). Among the 65 children of Group B, 20 (30.7%) went on to have one or more additional episodes. In conclusion, our study demonstrates that oral diazepam, given only when fever is present, is an effective means of reducing the risk of recurrences of FS. PMID- 15120685 TI - Reliability and validity of a qualitative and quantitative motor test for 5- to 6 year-old children. AB - Clumsiness in preschool children may be a precursor to impaired academic performance and psychological and developmental problems. It is assumed that in this age group especially the qualitative aspects (=pattern) of a movement reflect variations in motor development. Currently available motor tests for this age group, however, mostly objectify quantitative aspects of a movement alone and do not objectify qualitative aspects. The aim of this study was to develop a new, valid, and reliable tool (Maastricht's Motor Test (MMT)) to objectify qualitative and quantitative aspects of movement in 5- to 6-year-old children. The test covers Static Balance (14 items), Dynamic Balance (20 items), Ball Skills (eight items), and Diadochokinesis and Manual Dexterity (28 items). About 50% of the items measure qualitative aspects and 50% quantitative aspects of movements. In total 487 subjects were recruited from the first year of primary school. To validate the test, one school doctor's global judgment was used as a form of expert validity. Sensitivity and specificity were calculated for different cut off points. Intra-class correlation coefficients (ICC) of inter-rater (N = 42), intra-rater (N = 24), and test-retest (N = 43) agreement were determined. ICCs of the qualitative total score ranged from 0.61 to 0.95 and were comparable with those of the total quantitative score. The MMT can be used to objectify both qualitative and quantitative aspects of movements. The additional value of the qualitative observations has to be determined in children with various developmental problems. PMID- 15120686 TI - Developmental progress in Duchenne muscular dystrophy: lessons for earlier detection. AB - OBJECTIVE: To address the issue of diagnostic delay in Duchenne Muscular Dystrophy (DMD) using developmental data from a cohort of affected boys detected by newborn screening and data on the diagnostic pathways of a group of boys diagnosed clinically. DESIGN: Quantitative and semi-qualitative. SETTING: Primary care. SUBJECTS: 1. Cohort of boys diagnosed by newborn screening (NBS cohort), 2. Group of mothers whose sons were diagnosed clinically (LCD group) Interventions. NBS cohort: (a) Developmental milestones, (b) Griffiths assessment, (c) clinic letters, (d) family case studies. LCD group: semi-structured interview. MAIN OUTCOME MEASURE: 1. The effectiveness of previously proposed strategies for the earlier clinical diagnosis of DMD. 2. Diagnostic pathways of the LCD group. Factors contributing to diagnostic delay in the LCD group. RESULTS: 1. Previously proposed strategies for earlier diagnosis would have had limited effectiveness in detecting the NBS cohort. 2. Diagnostic delay continues because: (a) initial observations are usually non-specific and made by the family, (b) age of presentation and presenting symptoms are highly variable, (c) first concerns are usually expressed to the primary care team who are less likely to recognise the early indicators, (d) early locomotor symptoms could suggest an orthopaedic rather than a paediatric referral. CONCLUSIONS: The identification and implementation of an effective screening tool to reduce diagnostic delay is more complex than previously portrayed. In the light of this evidence service providers need to ask whether newborn screening is the only feasible solution to diagnostic delay. PMID- 15120687 TI - Arterial infarction caused by carotid artery dissection in the neonate. AB - Arterial stroke in a neonate caused by carotid artery dissection is rare. We report two cases, one with dissection at the level of the skull base, one just distal to the carotid bulb. Non-invasive techniques like MR angiography and sonography demonstrated the dissection accurately. MR imaging, especially the diffusion-weighted images, showed the extension and site of the cerebral infarction. In one case dissection could be suspected following vacuum and forceps extraction. In the other no obvious birth trauma was reported. In conclusion, in a neonate with clinical signs suggestive of cerebral infarction, dissection of the carotid artery should be considered. PMID- 15120688 TI - Acute hemiparesis in a boy with type 1 diabetes. AB - Unlike the case in adult-onset diabetes, neurologic complications in children with type 1 diabetes are uncommon. We report a case of a 14-year-old boy with type 1 diabetes who presented at our emergency unit with acute hemiparesis and intense headache without hypoglycaemia. Two hours after onset, the symptoms had subsided. It is the intention of the authors to increase the awareness of this association, whose prognosis is almost invariably good. PMID- 15120689 TI - Brachial plexus neuritis: is prognosis worse in children? PMID- 15120691 TI - Myometrial progesterone responsiveness and the control of human parturition. AB - OBJECTIVE: The goal of this review is to assess the body of literature addressing the mechanism of progesterone withdrawal in the control of human parturition and in particular the recent advances in testing the hypothesis that human parturition is initiated by decreased myometrial responsiveness to progesterone, ie, functional progesterone withdrawal. METHODS: Published studies of progesterone responsiveness of the pregnant human myometrium in the context of parturition control were reviewed. RESULTS: Advances in understanding the molecular basis for progesterone receptor (PR)-mediated control of progesterone responsiveness has led to the hypothesis that functional progesterone withdrawal in human parturition is mediated by specific changes in myometrial PR expression, function, or both. The human PR exists as two major subtypes, PR-A and PR-B. As PR-A represses progesterone actions mediated by PR-B, the extent of progesterone responsiveness is inversely related to the PR-A/PR-B expression ratio. In women, the onset of term labor is associated with a significant increase in the myometrial PR-A/PR-B expression ratio that may facilitate functional progesterone withdrawal. Interestingly, expression of the estrogen receptor-alpha (ERalpha) increases concordantly with the PR-A/PR-B expression ratio in nonlaboring myometrium. This finding indicates that functional estrogen activation and functional progesterone withdrawal are linked. CONCLUSION: Functional progesterone withdrawal in human parturition is likely mediated by an increase in the myometrial PR-A/PR-B expression ratio and possibly by modulation of coactivator and corepressor proteins. Functional progesterone withdrawal appears to induce functional estrogen activation. Thus, for most of pregnancy, progesterone may decrease myometrial estrogen responsiveness by inhibiting ERalpha expression. Such an interaction would explain why the human myometrium is refractory to the high levels of circulating estrogens for most of pregnancy. At term, functional progesterone withdrawal removes the suppression of ERalpha expression leading to an increase in ERalpha and a concomitant increase in myometrial estrogen responsiveness. Estrogen can then act to transform the myometrium to a contractile phenotype. This model explains why disruption of progesterone action alone triggers the full parturition cascade. The link between functional progesterone withdrawal and functional estrogen activation may be a critical mechanism for the endocrine control of human parturition. PMID- 15120692 TI - High concentrations of activin A in the peritoneal fluid of women with epithelial ovarian cancer. AB - OBJECTIVE: The aim of the present study was to evaluate the concentrations of activin A in the peritoneal fluid of women with epithelial (serous) ovarian cancer. METHODS: A group of 160 women was studied and divided in four subgroups as follows: 1) serous ovarian carcinoma (n = 32); 2) serous ovarian cystadenoma (n = 20); 3) endometriosis (n = 53); and 4) healthy controls (n = 55), including both fertile (n = 32) and postmenopausal women (n = 23). Specimens of peritoneal fluid were collected during surgical interventions, and activin A was quantified using a specific two-site enzyme immunoassay. RESULTS: Peritoneal fluid activin A concentrations in women with ovarian carcinoma were about five-fold higher than those found in the control group (median [interquartile range] = 7.60 [2.85 10.15] and 1.50 [1.00-2.50] ng/mL, respectively, P <.001). In contrast, the women with benign serous cystadenoma had peritoneal fluid activin A concentrations (1.50 [1.0-2.70] ng/mL) similar to those of the control group. High peritoneal fluid activin A levels (>2 multiples of the mean) distinguished carcinoma from cystadenoma with a sensitivity of 72% and a specificity of 80%. The follow-up of nine patients with stage IIIc ovarian cancer showed no apparent relationship between the peritoneal fluid activin A levels and overall survival. No significant difference in peritoneal fluid activin A concentrations between patients with endometriosis and control women was observed. CONCLUSION: Most women with serous ovarian carcinoma had high concentrations of activin A in the peritoneal fluid, supporting a possible role of this growth factor in ovarian cancer. PMID- 15120693 TI - Characterization of contractile activity and intracellular Ca2+ signalling in mouse myometrium. AB - OBJECTIVE: To characterize the contractile responses of mouse myometrium, the associated calcium (Ca2+) changes and the role of the sarcoplasmic reticulum (SR), and to better understand excitation contraction coupling in this tissue. METHODS: Strips of longitudinal myometrium were used, and Ca2+ was measured after loading with Indo-1. RESULTS: Intracellular Ca2+ transients, produced by Ca2+ entry, preceded phasic spontaneous contractions. Depolarization with high potassium concentration significantly increased the amplitude of the contractions and transformed the pattern of activity from phasic to tonic, with accompanying changes in intracellular Ca2+ concentration ([Ca2+]i). Oxytocin significantly stimulated contractile activity and [Ca2+]i above the level occurring spontaneously. Thus all forms of contractile activity were closely correlated with Ca2+. When the SR was emptied using a blocker of the SR calcium adenosinetriphosphatase, cyclopiazonic acid, spontaneous Ca2+ and force transients increased greatly in frequency and amplitude. Ryanodine, a blocker of Ca(2+)-induced Ca2+ release (CICR), did not impair activity. In the absence of external Ca2+, oxytocin was able to release Ca2+ from the SR through IP3 but produced only a small increase in force, demonstrating a requirement for Ca2+ entry as part of the mechanism of agonist action. CONCLUSION: Mouse myometrium, (1) produces contractile activity reflecting changes in [Ca2+]i irrespective of the stimulus, (2) has a significant SR Ca2+ content releasable by agonists but not CICR, (3) has an SR acting to inhibit spontaneous activity, and (4) behaves qualitatively similarly to human and rat myometrium in major aspects of excitation contraction coupling and is therefore a useful model tissue. PMID- 15120694 TI - Intracrine control of estrogen action in human gestational tissues at parturition. AB - OBJECTIVE: We examined whether estrogen action in human parturition is regulated by an intracrine mechanism mediated by target tissue expression of specific 17beta-hydroxysteroid dehydrogenase (17betaHSD) isozymes that interconvert estrone (E1) and estradiol (E2), such that the onset of labor is associated with an increase in local E2 bioavailability. METHODS: The extent of 17betaHSD-1, -2, 3, -4, -5, and -7 expression (measured by quantitative reverse transcriptase polymerase chain reaction) and the capacity to interconvert E1 and E2 were compared in amnion, chorion, placenta, decidua, and myometrium obtained from women at term before (n = 6) and after (n = 6) the onset of labor. RESULTS: In chorion, abundance of 17betaHSD-1 (converts E1 to E2) mRNA decreased 2.7-fold (P <.05) in association with labor onset. In myometrium, 17betaHSD-1 and 17betaHSD-4 (converts E2 to E1) mRNAs increased two-fold and five-fold, respectively, with the onset of labor (P <.05 for each). No other statistically significant labor associated change in 17betaHSD expression was observed. In chorion, 17betaHSD oxidative (E2 to E1) and reductive (E1 to E2) activities and the net E2 synthetic capacity increased with labor. In decidua, both activities decreased with the onset of labor, but there was no change in net E2 synthetic capacity. The capacity to interconvert E1 and E2 did not change in the other tissues. CONCLUSION: The increase in E2 synthetic capacity in the chorion might contribute to an increase in local estrogen bioactivity in association with the onset of labor. However, it cannot be explained by changes in 17betaHSD isozyme expression and is unlikely to account for the increased estrogen action at parturition. These data show that intracrine mechanisms based on 17betaHSD isozyme expression play a minor role, if any, in controlling estrogen action in gestational tissues during human parturition. PMID- 15120695 TI - Shift in expression of HLA-G mRNA spliceforms in pregnancies complicated by preeclampsia. AB - OBJECTIVE: Despite emerging data on the in vitro modulatory effects of trophoblast-associated human leukocyte antigen G (HLA-G), its in vivo function needs to be determined. Immunohistochemical studies show a decrease in protein expression of trophoblast HLA-G in preeclampsia. Such a decrease in protein might be the consequence of a shift in HLA-G mRNA spliceform patterns. In an exploratory pilot study we determined trophoblast HLA-G mRNA spliceform distribution in preeclampsia. METHODS: Placental samples were collected immediately after cesarean delivery from pregnancies complicated by preeclampsia or the syndrome hemolysis, elevated liver enzymes, and low platelet count (HELLP) and uncomplicated normotensive pregnancies as controls. HLA-G mRNA spliceform distribution was analyzed using a semiquantitative reverse transcriptase polymerase chain reaction procedure. RESULTS: Analysis of HLA-G spliceform distribution showed a significant increase in frequency of the G5 form encoding for a soluble HLA-G molecule in preeclampsia. This increase in G5 form was not found in pregnancies complicated by HELLP. CONCLUSION: The increased frequency in the expression of the HLA-G G5 spliceform may play a role in the pathophysiology of preeclampsia, in particular through a recently suggested effect of this soluble HLA-G molecule on remodeling of the spiral arteries. PMID- 15120696 TI - Polymorphisms of thrombophilic and vasoactive genes and severe preeclampsia: a pilot study. AB - OBJECTIVE: Carriage of thrombophilic and vasoactive polymorphic alleles has been associated with various pregnancy complications. The effect of carrying multiple polymorphisms is not known. We conducted a case-control study to determine the association between eight polymorphisms of thrombophilic and vasoactive genes and the risk of severe preeclampsia. METHODS: The following polymorphisms were analyzed by sequencing-on-chip-technology using solid-phase polymerase chain reaction on oligonucleotide microarrays: factor 5 (F5) Leiden, factor 2 (F2) prothrombin G20210A, plasminogen activator inhibitor (PAI)-1 4G/5G, nitric oxide synthase (NOS) 3 T768C, NOS 3 Glu298Asp, angiotensinogen (AGT) Met235Thr, estrogen receptor (ER) alpha Pvu II, and mineralcorticoid receptor (MLR) Ser810Leu. The study comprised 24 patients with severe preeclampsia and 24 controls from a cohort of consecutive white women treated at the Obstetrics Department of the University of Vienna Medical School. Genotypes were correlated with clinical data. RESULTS: The investigated polymorphisms did not influence the risk of severe preeclampsia independently. When separately considering the simultaneous carriage of multiple thrombophilic or vasoactive polymorphisms, neither the combined carriage of thrombophilic polymorphisms (F5 Leiden, F2 G20210A, PAI-1 4G/5G), nor the combined carriage of vasoactive polymorphisms (NOS 3 T768C, NOS 3 Glu298Asp, AGT Met235Thr) conferred an increased risk of severe preeclampsia. Cumulative genotype frequencies for at least two homozygous mutant genotypes, however, were nine of 24 (38%) and two of 24 (8%) for the study and control groups, respectively (P <.05). All of these nine women with severe preeclampsia had at least two homozygous mutant genotypes of four polymorphisms, ie, F5 Leiden, NOS 3 T768C, NOS 3 Glu298Asp, or ER alpha Pvu II. CONCLUSION: Our data fail to document an independent significant influence of the investigated polymorphisms on the risk of severe preeclampsia. In an attempt to build a multigenetic model of severe preeclampsia, the combination of F5 Leiden, NOS 3 T768C, NOS 3 Glu298Asp, and ER alpha Pvu II was the most effective combination to predict the presence of severe preeclampsia in this small series of white women. PMID- 15120697 TI - Polymorphism at codon 72 of the p53 gene is not associated with endometriosis in a Japanese population. AB - OBJECTIVE: Endometriosis is inherited as a complex trait, which means that multiple susceptibility genes interact with each other and the environment to produce the phenotype. Previous studies have implicated p53, a tumor suppressor gene, as a factor in the development of the disease. In a Japanese population, we investigated the frequency of the p53 polymorphism in women affected with endometriosis. METHODS: We compared the distribution of the p53 codon 72 polymorphism in endometriosis cases (n = 111) and population controls consisting of female neonates (n = 180) by using polymerase chain reaction restriction fragment-length polymorphism analysis in a Japanese population. RESULTS: The frequencies of the three p53 genotypes, Arginine (Arg)/Arg, Arg/Proline (Pro), and Pro/Pro in controls were 39.4%, 41.7%, and 18.9 %, respectively. The crude genotype frequencies in the endometriosis cases were similar to those of the controls (35.2%, 48.6%, and 16.2%, respectively). Using the Arg/Arg genotype as the reference, the odds ratios of the Arg/Pro and Pro/Pro genotypes were 1.30 (95% confidence interval [CI] 0.72-1.86, P =.33) and 0.96 (95% CI 0.47-1.94, P =.91), respectively. Thus, there were no significant differences in the frequency of the p53 codon 72 polymorphism between endometriosis cases and controls in this population. The endometriosis cases with severe disease only were also evaluated, but no significant difference was observed in the frequency of the polymorphism between this subgroup and the controls. CONCLUSION: Our findings suggest that the p53 codon 72 polymorphism is unlikely to be associated with endometriosis in Japanese women. PMID- 15120698 TI - Androgen receptor gene cytosine, adenine, and guanine trinucleotide repeats in patients with endometriosis. AB - OBJECTIVE: A genetic variation in the androgen receptor (AR) has been associated with the risk of developing endometriosis. The AR gene is located on the X chromosome and contains a highly polymorphic trinucleotide repeat (cytosine, adenine, and guanine: CAG) in its first exon, whose length and methylation pattern affect both AR expression and function. Thus, we sought to further investigate the potential association between endometriosis and the AR-CAG polymorphism. METHODS: Genomic DNA was obtained from a consecutive series of 197 white Italian women of reproductive age who underwent laparoscopy for benign gynecologic pathologies. Molecular analysis of AR-CAG repeats was performed by polymerase chain reaction amplification and Genescan evaluation. The pattern of CAG repeat distribution was compared between subjects with and without endometriosis. RESULTS: Endometriosis was documented in 105 women (stage I-II in 33 women and stage III-IV in 72 women). We found no difference in the number of AR-CAG repeats between women with endometriosis and controls. The CAG repeat length ranged from eight to 27 (mean +/- standard deviation, 17.4 +/- 1.9) for endometriosis patients and from 11 to 27 (mean +/- standard deviation, 17.4 +/- 2) for controls. Moreover, no association was found between AR gene polymorphisms and the various clinical manifestations of the disease. CONCLUSION: We conclude that AR-CAG repeat length does not constitute an important factor for the genetic predisposition to endometriosis. PMID- 15120699 TI - Effects of epidermal growth factor/hydrocortisone on the growth and differentiation of human ovarian surface epithelium. AB - OBJECTIVE: Ovarian surface epithelium (OSE), the precursor of the epithelial ovarian carcinomas, has limited growth potential in culture. Epidermal growth factor+hydrocortisone (EGF+HC) enhances its growth but induces epitheliomesenchymal transition (EMT). This study was undertaken to define the effects of EGF+HC and their reversibility, to optimize growth-promoting media, and to relate OSE phenotypes in vitro to physiologic states in vivo. METHODS: OSE was cultured in media 199/MDCB105 or EBM (Clonetics) with 2% or 10% fetal bovine serum with or without 10 ng/mL EGF, 1.0 microg/mL HC, and 1.0 microg/mL bovine brain extract. Growth rates and growth potentials (population doublings [PD] to senescence) were defined, and growth patterns and expression of keratin and collagen types III and IV were compared with the ovarian cancer cell lines OVCAR3 and SKOV3. RESULTS: EGF+HC increased growth potentials from 12-14 PD to 40-42 PD and reduced PD time from 53 hours to 20 hours. Without EGF+HC, OSE cells remained uniformly epithelial. EGF+HC induced EMT (mesenchymal shapes, reduced keratin, and production of collagenous extracellular matrix), but the EMT response varied greatly among OSE from different women. EMT was reversed over 1-2 weeks by subculture into EGF+HC-free medium in passage 1, but inconsistently thereafter. EGF+HC had no effect on the differentiation of ovarian carcinoma lines. CONCLUSION: The phenotype of intact OSE in vivo is most closely reproduced in media without EGF+HC. EGF+HC enhances growth but initiates EMT, which likely mimics a repair response. Variations in EGF+HC-induced phenotypes point to the existence of OSE subpopulations with differing responsiveness to growth factors or steroids, which may relate to their susceptibility to malignant transformation. PMID- 15120700 TI - Apoptosis-based evaluation of chemosensitivity in ovarian cancer patients. AB - OBJECTIVE: Induction of apoptosis in target cells is a key mechanism by which chemotherapy induces cell killing. We have established an in vitro system for determining the chemosensitivity of epithelial ovarian cancer cells to carboplatin and paclitaxel (Taxol). Practical assays to predict the likelihood of individual tumor sensitivity are needed to facilitate the choice of adequate treatment. We sought to determine whether epithelial ovarian cancer cells (EOC) collected from the ascites fluid of patients known to be clinically chemosensitive or chemoresistant to carboplatin and paclitaxel would show a similar response to chemotherapeutic drugs after in vitro treatment. METHODS: Thirteen patients with stage III and IV ovarian cancer treated with carboplatin and paclitaxel were studied. Caspase-3 activation was used as a surrogate marker for activation of chemotherapy-induced programmed cell death. We compared the in vitro apoptotic response to the clinical response of the patients from whom the tumor cells were isolated. Clinical sensitivity was defined as no evidence of disease recurrence for 6 months after optimal debulking surgery and completion of chemotherapy. RESULTS: Of seven chemosensitive patients, five cell samples treated in vitro had increased caspase-3 activity in response to both carboplatin and paclitaxel. Five of six chemoresistant cases did not show caspase-3 activity in response to only one or to neither agent. CONCLUSION: Quantifiable markers of apoptosis such as caspase-3 activation have the potential to predict the clinical response to chemotherapy. Application of this assay in clinical laboratories could optimize the potential for efficient treatment and avoid the toxicities of ineffective drugs. PMID- 15120701 TI - Clinical experience with thioctacid (thioctic acid) in the treatment of distal symmetric polyneuropathy in Korean diabetic patients. AB - OBJECTIVE: This open-label study was performed to evaluate the efficacy and safety of oral treatment with the antioxidant alpha-lipoic acid (ALA, thioctic acid) in Korean diabetic patients with distal symmetric polyneuropathy (DSP). SUBJECTS AND METHODS: Thioctic acid was administered orally using 600 mg once daily for 8 weeks in 61 diabetic patients with symptomatic polyneuropathy. Neuropathic symptoms (pain, burning sensation, paresthesia, and numbness) were scored at baseline as well as at 4 and 8 weeks following treatment. In addition, neurological assessment was carried out before and after 8 weeks of treatment, and an overall evaluation was performed at the end of treatment. The primary endpoint was the response rate after 8 weeks of treatment, defined as an improvement in the Total Symptom Score (TSS) of > or =30%. RESULTS: Efficacy was evaluated in 38 patients who had completed the study according to the protocol. Safety was evaluated in all 61 patients who had taken the study medication. Fasting blood glucose and HbA(1)c did not change during the study. The response rate after 8 weeks was 71.4%. At 4 weeks, the response rate was 47.4%. The TSS significantly decreased at 4 weeks, which decreased further at 8 weeks (P<.05). All the individual scores for neuropathic symptoms (pain, burning sensation, paresthesia, and numbness) were also significantly reduced at 4 weeks and further decreased at 8 weeks (P<.05). The duration of diabetes, severity and duration of diabetic polyneuropathy, and all the other demographic and metabolic parameters did not demonstrate an effect on the response rate. The parameters of neurological assessment (ankle reflexes, pin-prick test, 10-g monofilament test) and quantitative sensory tests (vibration, warm and cold sensation) were not influenced by 8 weeks of treatment with 600 mg of oral thioctic acid per day. Overall efficacy rated as "good/fair" was 86.8% by the physician and 76.3% by the patients at the end of an 8-week treatment period. Eleven episodes (18.0%) of adverse events (possibly, probably, definitely related) were reported in seven patients (11.5%). CONCLUSION: These findings indicate that oral treatment with thioctic acid at a dose of 600 mg/day for 8 weeks improved symptoms of polyneuropathy in Korean diabetic patients without causing serious adverse events. PMID- 15120702 TI - Detection of subsequent episodes of gestational diabetes mellitus: a need for specific guidelines. AB - Guidelines for detection of individuals with gestational diabetes mellitus (GDM) indicate that glucose testing for women with a history of GDM should occur as soon as feasible with retesting of an initially negative screen to occur between the 24th and 28th week of gestation. The aim of this study was to evaluate medical records for individuals enrolled in a GDM management program that presented with two subsequent pregnancies with GDM and to determine if more specific guidelines for detection are needed. Records (n=60) from both pregnancies were reviewed for gestational age at enrollment, delivery, and when insulin was started, infant birth weights and complications (e.g., hypoglycemia), and maternal complications (e.g., emergency cesarean section). Over half [33/60 (55%)] of the women required insulin during both pregnancies, while 16.7% (10/60) required insulin during the second enrollment for GDM but not the first. For those requiring insulin during both pregnancies, 88% (29/33) required it earlier during the subsequent pregnancy (31.5+/-2.7 vs. 21.6+/-8.4 weeks of gestation, P<.001). During the subsequent pregnancy, approximately 1/2 of the women requiring insulin needed it before the 24th week of gestation while 1/3 required it by the 15th week. Also during the subsequent pregnancy, neonate birth weights declined (3494+/-521 vs. 3356+/-515 g, P<.05) and there were fewer complications. Given that approximately 70% of the women required insulin therapy during a subsequent GDM pregnancy and that this therapy was on average necessary by the 22nd week of gestation, we recommend that specific guidelines be established with a definitive time frame determined for the detection of repeat episodes of GDM. PMID- 15120703 TI - Endothelial dysfunction in patients with type 2 diabetes and the effects of thiazolidinedione antidiabetic agents. AB - Endothelial dysfunction is increasingly recognised as a key event in the pathogenesis of atherosclerosis, which occurs in association with insulin resistance early in the course of type 2 diabetes mellitus (T2DM). Thiazolidinediones (TZDs), such as rosiglitazone, are a class of oral antidiabetic agents that act primarily as insulin sensitisers, reducing insulin resistance with associated improvements in glycemic control. Available data indicate that thiozolidinediones also have beneficial effects on numerous markers of endothelial function and profound antiinflammatory activity, indicative of potential antiatherogenic activity. These effects may be of considerable clinical significance if sustained during long-term therapy, given the morbidity and mortality associated with atherosclerosis in T2DM patients. PMID- 15120705 TI - Comparison of microbial findings and resistance to antibiotics between transplant patients, patients on hemodialysis, and other patients with the diabetic foot. AB - Infectious complications of the diabetic foot may be influenced by impaired renal function and by immunosuppression therapy. AIMS: To assess differences in microbial findings and resistance to antibiotics between transplant recipients, hemodialysis patients, and other patients with the diabetic foot. METHODS: 207 patients treated in the foot clinic for diabetic ulcers from 12/1998 to 12/1999 were included into this retrospective study. Patients were divided into three groups (transplant, dialysis, and other patients). Occurrence of individual bacterial species and resistance to antibiotics was compared between study groups. RESULTS: Study groups did not differ significantly in ulcer grades defined by the Wagner classification or in the mean number of pathogens per patient. The prevalence of individual microorganisms did not differ between the study groups. However, the study groups differed significantly in the occurrence of microbial resistance to antibiotics. Transplant patients had more frequently Staphylococcus aureus resistant to oxacillin (P<.01), imipenem (P<.01), co trimoxazole (P<.01), Enterococcus species resistant to ampicillin (P<.01), piperacillin (P<.01), and dialysis patients had more frequently Pseudomonas species resistant to piperacillin (P<.05) and cefpirom (P<.05) in comparison with the other two groups. CONCLUSIONS: Transplant patients had significantly more resistant microorganisms in comparison with dialysis and other patients with the diabetic foot. Empiric antibiotic selection based on general population data should be modified in transplant patients with diabetic foot according to actual susceptibility to antibacterial drugs. PMID- 15120706 TI - A brief, regular, proactive telephone "coaching" intervention for diabetes: rationale, description, and preliminary results. AB - Telephone-delivered interventions (TDIs) represent a potentially cost-effective method to increase medical adherence. TDIs for diabetes patients have typically been delivered by nurses or computerized telephone messaging. Psychology undergraduates, however, are less costly than nurses, have a strong background in behavioral science, and provide the personal relationship missing with computerized contact. This paper presents the rationale for and description of a brief, regular, proactive telephone intervention designed to be delivered by psychology undergraduates (i.e., paraprofessionals). "Coaches" administer a 15 min telephone intervention weekly for 3 months and biweekly for 3 additional months. Guided by a semistructured protocol that focuses on behavioral goals, coaches provides support, collaborative problem-solving, and apply basic cognitive-behavioral techniques. Results from a pilot study on type 1 diabetes patients are presented. This preliminary evidence suggests that the program is feasible, acceptable to a large majority of patients, and effective in reducing HbA1c levels. PMID- 15120704 TI - Putative role of polymorphisms in UCP1-3 genes for diabetic nephropathy. AB - Increased production of reactive oxygen species (ROS) has been suggested as a cause of diabetic complications. Uncoupling proteins (UCPs) have been ascribed a role in reducing the formation of ROS, and genetic variation in genes encoding for UCPs could thus be putative candidate genes for diabetic nephropathy. To test this hypothesis we searched for association between the A-->G (-3862) variant in UCP1, the insertion/deletion (I/D) polymorphism in exon 8 in UCP2, and the C-->T (-55) polymorphism in UCP3 and diabetic nephropathy in 218 diabetic patients with normal urinary albumin excretion rate (AER), 216 with micro- or macroalbuminuria, and in 106 control subjects without a family history of diabetes. We did not find any association between the different polymorphisms and diabetic nephropathy, nor did we observe any difference in AER among carriers of different UCP1-3 genotypes. We could, however, confirm the reported association between BMI and the UCP3 -55 C-->T polymorphism; patients carrying the T allele had higher BMI than patients homozygous for the C allele (26.4+/-4.2 vs. 25.3+/-4.3 kg/m(2); P=.01). We conclude that studied polymorphisms in the UCP1-3 genes do not play a major role in the development of micro- or macroalbuminuria in Scandinavian diabetic patients. PMID- 15120707 TI - Diabetic rats treated by low molecular weight heparin OP 2123/parnaparin: morphological changes in the kidney and heart. AB - AIM: Early morphological alterations in the rat kidney and heart due to experimentally induced diabetes are described in order to evaluate the possible therapeutic role of low molecular weight heparin (LMWH; OP 2123/parnaparin). METHODS: Our findings concern the alterations observed in the rat kidney and heart because these are the organs (together with the retina) mainly involved in the early morphological angiopathic modifications associated with diabetic damage of organs and tissues. In diabetic animals treated with LMWH, the Periodic Acid Schiff (PAS) reaction showed a slight decrease when compared with the diabetic control group. Photographs were submitted to the quantitative analysis of images using a Quantimet 500 Image Analyzer (Leica) equipped with specific software. The following parameters were measured: (1) total area occupied by alkaline phosphatase (AP)-positive capillaries; (2) number and diameter of AP-positive capillaries; (3) distribution and total area occupied by PAS-positive structures (related to the intensity of the reaction resulting from the different amount of mucopolysaccharides). RESULTS: LMWH treatment is efficient in preventing these modifications, above all in the kidney. The histological study of the heart and kidney shows no significant, relevant alterations. However, the histological study of the mucopolysaccharides in diabetic animals highlighted a tendency for the heart to accumulate these substances. LMWH treatment only modestly reduced this accumulation. CONCLUSIONS: Previous evidence demonstrating a beneficial effect of therapy based on heparan sulphate proteoglycans and/or other heparin like substances in insulin-dependent diabetes mellitus seems to be confirmed by our experimental results in different organs of adult rats. In fact, parnaparin treatment is effective (in our experience) for ameliorating the morphological pattern observed early in some diabetic tissues of rats and, above all, in the kidney. PMID- 15120708 TI - Severe and disabling diabetic autonomic neuropathy: a case report. AB - We report a case of a woman with several and severe disabling manifestations of autonomic neuropathy in whom reasonable quality of life was established by combining continuous insulin infusion, jejunal feeding, colostomy and bladder self-catheterisation. We discuss the prevalence rates, pathophysiology, management and prognosis of this disabling condition. PMID- 15120709 TI - Pharmacological prevention of diabetic cataract. AB - Cataract--opacification of the lens--is closely related to diabetes as one of its major late complications. This review deals with three molecular mechanisms that may be involved in the development of diabetic cataract: nonenzymatic glycation of eye lens proteins, oxidative stress, and activated polyol pathway in glucose disposition. Implications resulting from these mechanisms for possible pharmacological interventions to prevent diabetic cataract are discussed. The article reviews research on potential anticataract agents, including glycation inhibitors, antioxidants, and aldose reductase inhibitors. Information on possible benefits of putative anticataract agents comes from a variety of approaches, ranging from laboratory experiments, both in vitro and in vivo, to epidemiological studies in patients. PMID- 15120710 TI - Incorporation of arachidonic and linoleic acid hydroperoxides into cultured human umbilical vein endothelial cells. AB - The current study assessed the differential incorporation of 12 hydroperoxyeicosatetraenoic acid (12-HPETE), arachidonic acid (AA), 12 hydroxyeicosatetraenoic acid (12-HETE) and the linoleic acid (LA) oxidation products, 13-hydroxyoctadecadienoic acid (13-HODE) and 13 hydroperoxyoctadecadienoic acid (13-HPODE), into human umbilical vein endothelial cells (HUVEC). Approximately 80-90% of AA (10(-8)-10(-5)M) and 80% of LA (10(-8) 10(-5)M) were incorporated into HUVEC within 12h, while less than 50% of the hydroxy metabolites (12-HETE, 12-HPETE, 13-HODE, 13-HPODE) were incorporated into HUVEC over 48h. Further, treatment of HUVEC with either 12-HPETE or 13-HPODE (concentrations of 10(-5)M) had no effect on cell number at a 48h time point when compared with control. These results demonstrate that exogeneous hydroxy metabolites are incorporated into HUVEC to a lesser degree than were endogenous fatty acids. Further, we speculate that 12-HPETE and 13-HPODE are rapidly metabolized to substances without significant cytotoxic effects. PMID- 15120711 TI - The prostacyclin analogue beraprost sodium prevents development of arterial stiffness in elderly patients with cerebral infarction. AB - Prostacyclin (PGI(2)) inhibits platelet aggregation, smooth muscle cell proliferation, and vasoconstriction. Arterial stiffness assessed by pulse wave velocity (PWV) predicts mortality in various cardiovascular diseases. To study the preventive effects of a prostacyclin analogue, beraprost sodium, on arterial PWV values in elderly patients with cerebral infarction. Forty-four patients with a history of cerebral infarction received beraprost sodium (120 microg/day p.o.) or no beraprost sodium (control) for 3 months. Arterial PWV and ankle brachial indices (ABI) were determined prior to starting the medication and after 3 months of medication. Initially, there were no differences in age, blood pressure, and body mass index. Further, PWV or ABI did not differ between the beraprost sodium group (n = 22) and the control group (n = 22). After 3 months, PWV in beraprost sodium group was significantly reduced (-123 +/- 282) when compared with the control group (147 +/- 274)(P = 0.006). ABI was not significantly different when comparing the two groups at 3 months. Long-term administration of beraprost sodium prevents the decline in arterial biomechanics in elderly patients with cerebral infarction. PMID- 15120712 TI - Depression and adipose polyunsaturated fatty acids in the survivors of the Seven Countries Study population of Crete. AB - The purpose of the present study was to investigate the relation between adipose tissue polyunsaturated fatty acids, an index of long-term or habitual fatty acid dietary intake and depression. The sample consisted of 150 elderly males from the island of Crete. The subjects were survivors of the Greek Seven Countries Study group. The mean age was 84 years. The number of subjects with complete data on all variables studied was 63. Subjects were examined by the Preventive Medicine and Nutrition Clinic of the University of Crete. Depression was assessed through the use of the short form of the Geriatric Depression Scale (GDS-15). Depression correlated negatively with adipose tissue alpha-linolenic acid (C18:3n-3). Depressed subjects had significantly reduced (-10.5%) adipose tissue C18:3n-3 levels than non-depressed subjects. The observed negative relation between adipose tissue C18:3n-3 and depression, in the present study, appears to indicate increasing long-term dietary C18:3n-3 intakes with decreasing depression. This agrees with findings of other studies indicating an inverse relation between depression and consumption of fish and n-3 polyunsaturated fatty acids. This is the first literature report of a relation between adipose tissue C18:3n-3 and depression. Furthermore, this is the first report of a relation between adipose PUFA and depression in an elderly sample. Depression has been reported to be associated with elevated cytokines, such as, IL-1, IL-2, IL-6, INF-gamma and INF alpha. Fish oil and omega-3 fatty acids, on the other hand, have been reported to inhibit cytokine production. The observed negative relation between adipose C18:3n-3 and depression, therefore, may stem from the inhibiting effect of C18:3n 3 or its long-chain metabolites on cytokine synthesis. PMID- 15120713 TI - Microcirculation and tolerability following i.v. infusion of PGE1 and iloprost: a randomized cross-over study in patients with critical limb ischemia. AB - In a randomized cross-over study, the effect of PGE(1) and iloprost on microcirculation as well as the tolerability was investigated in 36 patients with peripheral arterial occlusive disease stage III and IV according to Fontaine. Patients received PGE(1) and iloprost by single 3-h i.v. infusions on two different days at doses recommended by the manufacturers or in previous studies (PGE(1): first hour 20 microg, next 2h 30 microg each. Iloprost: first hour 0.5 ng/kg/min, next 2h 1.0 ng/kg/min). Transcutaneous oxygen pressure (tcPO(2)) values increased much more with PGE(1). Median tcPO(2) increase over baseline 30 min after the end of infusion was 9 and 2 mmHg for PGE(1) and iloprost, respectively, corresponding to median AUC differences from baseline of 1050 and 210 min mmHg. Because of its exploratory character, the study was not powered to test for significance. Adverse effects occurred in 19.4% (PGE(1)) and 30.6% (iloprost) of patients. Dose reduction was required in 3 patients receiving iloprost (hypotension, nausea, irritation of the infused vein), and in none receiving PGE(1). PMID- 15120714 TI - Phytosterols decrease prostaglandin release in cultured P388D1/MAB macrophages. AB - Cardiovascular disease (CVD) remains the leading cause of death in Western societies. Atherosclerosis is a major cardiovascular related disorder that is responsible for 50% of all mortality in the United States. Several epidemiological studies suggest that consumption of a plant-based diet is associated with a decreased incidence of cardiovascular abnormalities. Phytosterols, especially beta-sitosterol, are plant sterols that have been shown to exert protective effects against cardiovascular diseases as well as many types of cancer. Monocyte/macrophage cells are involved with the inflammatory process. Accumulation of these cells in arteries is one of the initial events leading to atherosclerosis. Macrophages are capable of supplying the atherosclerotic vessel with substantial amounts of prostaglandins. Prostaglandins have been shown by numerous studies to play a key role in the atherosclerosis process. They can affect platelet aggregation, vasodilation or constriction of blood vessels, and the adherence of monocytes to the vessel walls. The purpose of this study was to examine the effect of phytosterols on the release of PGE(2) and PGI(2) from lipopolysaccharide (LPS)-stimulated P388D(1)/MAB macrophage cells. P388D(1)/MAB cells were supplemented with 16 microM cholesterol, beta-sitosterol or campesterol using cyclodextrin as a vehicle. Phytosterol supplementation led to a significant decrease in cellular growth at various time points throughout a 7-day treatment period, especially after 3 days of treatment. Macrophages incorporated the supplemented phytosterols into their membranes which accounted for 26% of total membrane sterols. Cholesterol supplementation at 16 microM however, had no effect on membrane sterols. Supplementation with 16 microM concentration of beta sitosterol or campesterol resulted in a significant inhibition of PGE(2) and PGI(2) release from macrophage cells as compared to the vehicle control. Of the two phytosterols, beta-sitosterol supplementation exhibited a greater inhibitory effect. PGE(2) release was decreased 68% by beta-sitosterol and 55% by campesterol, while cholesterol supplementation was not as effective, as it led to a 37% decrease. Similarly, release of PGI(2) from macrophages was inhibited 67% by beta-sitosterol and 52% by campesterol treatment, while enrichment of the cells with cholesterol, led to a 35% decrease in PGI(2) release. The decrease in prostaglandin release was not due to alteration in the expression of cPLA(2) and COX-2 enzymes which suggests that alterations in the activities of these enzymes may be responsible for the observed changes in prostaglandin release. It was concluded that phytosterol incorporation into macrophages may offer protection from atherosclerosis by reducing their prostaglandin release and thus slowing down the atheroma development. PMID- 15120715 TI - Spice phenolics inhibit human PMNL 5-lipoxygenase. AB - A wide variety of phenolic compounds and flavonoids present in spices possess potent antioxidant, antimutagenic and anticarcinogenic activities. We examined whether 5-lipoxygenase (5-LO), the key enzyme involved in biosynthesis of leukotrienes is a possible target for the spices. Effect of aqueous extracts of turmeric, cloves, pepper, chili, cinnamon, onion and also their respective active principles viz., curcumin, eugenol, piperine, capsaicin, cinnamaldehyde, quercetin, and allyl sulfide were tested on human PMNL 5-LO activity by spectrophotomeric and HPLC methods. The formation of 5-LO product 5-HETE was significantly inhibited in a concentration-dependent manner with IC(50) values of 0.122-1.44 mg for aqueous extracts of spices and 25-83 microM for active principles, respectively. The order of inhibitory activity was of quercetin>eugenol>curcumin>cinnamaldehyde>piperine>capsaicin>allyl sulfide. Quercetin, eugenol and curcumin with one or more phenolic ring and methoxy groups in their structure showed high inhibitory effect, while the non-phenolic spice principle allyl sulfide showed least inhibitory effect on 5-LO. The inhibitory effect of quercetin, curcumin and eugenol was similar to that of synthetic 5-LO inhibitors-phenidone and NDGA. Moreover, the inhibitory potency of aqueous extracts of spice correlated with the active principles of their respective spices. The synergistic or antagonistic effect of mixtures of spice active principles and spice extracts were investigated and all the combinations of spice active principles/extracts exerted synergistic effect in inhibiting 5-LO activity. These findings clearly suggest that phenolic compounds present in spices might have physiological role in modulating 5-LO pathway. PMID- 15120716 TI - The influence of different combinations of gamma-linolenic, stearidonic and eicosapentaenoic acids on the fatty acid composition of blood lipids and mononuclear cells in human volunteers. AB - This study set out to identify whether stearidonic acid (18:4n-3; STA) can be used to increase the eicosapentaenoic acid (20:5n-3; EPA) content of plasma lipids and cells in humans and to understand more about the effects of increased consumption of gamma-linolenic acid (18:3n-3; GLA), STA and EPA in humans. Healthy young males were randomised to consume one of seven oil blends for a period of 12 weeks (9g oil/day) (n = 8-12 subjects/group). Palm oil, sunflower oil, an EPA-rich oil, borage oil (rich in GLA), and Echium oil (rich in STA) were blended in various combinations to generate a placebo oil and oils providing approximately 2g GLA + STA + EPA per day, but in different combinations. Blood was collected at 0, 4, 8 and 12 weeks and the fatty acid compositions of plasma triacylglycerols, cholesteryl esters and phospholipids and of peripheral blood mononuclear cells (PBMCs) determined. Significant effects were observed with each lipid fraction. Neither STA nor its derivative 20:4n-3 appeared in any of the lipid fractions studied when STA (up to 1g/day) was consumed. However, STA (1g/day), in combination with GLA (0.9 g/day), increased the proportion of EPA in some lipid fractions, suggesting that STA-rich plant oils may offer a novel means of increasing EPA status. Furthermore, this combination tended to increase the dihomo-gamma-linolenic acid (20:3n-6; DGLA) content of PBMCs, without an increase in arachidonic acid (AA) (20:4n-6) content. EPA consumption increased the EPA content of all lipid fractions studied. Consumption of GLA (2g/day), in the absence of STA or EPA, increased DGLA content with a tendency to increase AA content in some fractions. This effect was prevented by inclusion of EPA in combination with GLA. Thus, this study indicates that STA may be used as a precursor to increase the EPA content of human lipids and that combinations of GLA, STA and EPA can be used to manipulate the fatty acid compositions of lipid pools in subtle ways. Such effects may offer new strategies for manipulation of cell composition in order to influence cellular responses and functions in desirable ways. PMID- 15120717 TI - From bench to the clinic: gamma-linolenic acid therapy of human gliomas. AB - Malignant gliomas are among the most devastating of cancers and are a major cause of mortality in a young population with a median survival time of 9 months following cytoreductive surgery, radiotherapy and chemotherapy. Recent studies showed that polyunsaturated fatty acids especially gamma-linolenic acid (GLA) have selective tumoricidal action especially against malignant glioma cells both in vitro and in vivo. Limited open label clinical studies showed that intratumoral injection/infusion of GLA is safe and effective against malignant gliomas. In view of this, large-scale, double blind studies are needed to establish the usefulness of GLA in the treatment of malignant brain tumors. PMID- 15120718 TI - A new respiratory fluoroquinolone, oral gemifloxacin: a safety profile in context. AB - Gemifloxacin is a broad-spectrum quinolone antibacterial with enhanced potency against Gram-positive bacteria, including multi-drug resistant Streptococcus pneumoniae, and retained potency against Gram-negative bacilli and bacterial strains resistant to other antibiotics. It has proven particularly effective in respiratory and urinary tract infection. This review presents safety data from 6775 patients included in clinical trials, receiving either the recommended 320 mg once daily oral dose of gemifloxacin, or standard dose of other quinolones, macrolides or beta-lactams (n = 5248). Studies in healthy volunteer and special populations are also reported. Adverse experiences (AEs) were observed in 44.7% of gemifloxacin-treated patients and 47.5% of those who received comparator drugs. Mild gastro-intestinal adverse drug reactions (ADRs) (diarrhoea 5.1%, nausea 3.9%) predominated. Rash, usually maculo-papular and in no case proceeding to more severe eruptions, was observed in 3.6% of those receiving gemifloxacin. A higher incidence of rash (>20%) was observed in young women and was the subject of further study. Adverse drug reactions suspected or probably related to treatment occurred in 17.4% of patients receiving gemifloxacin and in 20% of those receiving comparator antibiotics. Diarrhoea and nausea were experienced by 3.6 and 2.7%, respectively, of gemifloxacin-treated patients (4.6 and 3.2% of comparators), rash by 2.8% (0.6% of comparators) and headache by 1.2% (1.5% of comparators). Gemifloxacin-related vomiting (0.9%), dizziness (0.8%) and taste perversion (0.3%) were uncommon. Treatment discontinuation followed one or more adverse drug reactions in 2.2% of gemifloxacin-treated patients (0.9% due to rash) and 2.1% of comparator-treated patients. A total of 63 deaths (33 receiving gemifloxacin) occurred in the trial population: none were considered related to treatment. A slight prolongation in QT interval (2.56 ms (S.D. +/-24.5)) was observed in gemifloxacin-treated patients: no cardiac arrhythmias were reported. There was a low incidence of liver function tests (LFTs) classified as of potential clinical concern: gemifloxacin (0.4-1.2%), comparators (0.2-1.3%). Serious adverse events (SAEs), occurring during but not necessarily related to therapy, occurred in 3.6% of gemifloxacin-treated patients (4.3% of comparators). SAEs related to treatment agents were rare (0.4% in each group) and included rash (0.1%) and elevated liver enzymes (<0.1%). Gemifloxacin was well tolerated by the elderly, those with renal or hepatic impairment and when co-administered with omeprazole, digoxin, theophylline, warfarin (with which there were no significant interactions) and Maalox. In conclusion, gemifloxacin 320 mg once daily demonstrated a favourable safety and tolerability profile similar to that of comparator antibiotics, including other quinolones. PMID- 15120719 TI - Effects of four antiviral substances on lethal vaccinia virus (IHD strain) respiratory infections in mice. AB - Intranasal infection of BALB/c mice with the IHD strain of vaccinia virus was found to cause pneumonia, profound weight loss and death. Cidofovir, hexadecyloxypropyl-cidofovir (HDP-CDV), the diacetate ester prodrug of 2-amino-7 [(1,3-dihydroxy-2-propoxy)methyl]purine (HOE961), and ribavirin were used to treat the infections starting 24h after virus exposure. Single intraperitoneal (i.p.) cidofovir treatments of 100 and 30 mg/kg led to 90-100% survival compared with no survivors in the placebo group, whereas a 10 mg/kg dose was ineffective. The 100 mg/kg treatment reduced lung and snout virus titres on day 3 of the infection by 20- and 8-fold, respectively. Mean arterial oxygen saturation levels in these two cidofovir treatment groups were significantly higher than placebo on days 4 through 6 of the infection, indicating an improvement in lung function. Effects of cidofovir on viral pathogenesis were studied on days 1, 3 and 5 of the infection, and demonstrated statistically significant reductions in lung consolidation scores, lung weights, lung virus titre and snout virus titres on days 3 and 5. Cidofovir treatment also reduced virus titres in other tissues and body fluid, including blood, brain, heart, liver, salivary gland and spleen. HDP CDV was given by oral gavage at 100, 50 and 25mg/kg doses one time only, resulting in 80-100% survival. Lower daily oral doses of 10 and 5mg/kg per day given for 5 days protected only 30% of animals from death. Oral doses (100, 50 and 25 mg/kg per day) of HOE961 for 5 days protected all animals, whereas equivalent oral doses of ribavirin were completely ineffective. The rapidity of recovery from weight loss during the infection was a function of dose of compound administered. These data indicate the utility of parenteral cidofovir, oral HDP CDV and oral HOE961 in treating severe respiratory infections caused by this virus. PMID- 15120720 TI - Changes before and after a policy to restrict antimicrobial usage in upper respiratory infections in Taiwan. AB - The Bureau of National Health Insurance (BNHI) of Taiwan issued a new reimbursement regulation effective from 1 February 2001 forbidding the use of antimicrobials in ambulatory patients with upper respiratory infections (URI) without evidence of bacterial infection. We evaluated the effect of this regulation by analysing changes in the types of infections diagnosed and the amount of antibiotics prescribed in 1999, 2000 and 2001. Between 1999 and 2001, antimicrobials for respiratory infections decreased from 18.0 to 9.97 DDD/1000 per day or by 44.6% (P=0.0000+). Antimicrobials for URI decreased from 8.32 in 1999 to 3.28 DDD/1000 per day in 2001 or by 60.6% (P=0.0000+); from 2000 to 2001 the decrease was 55.8%. Reduction of antimicrobials for URI from 1999 to 2001 accounted for 62.8% of the reduction of antimicrobials in respiratory infections or 51.3% of the total reduction of antimicrobials. Reduction in aminopenicillins was responsible for most of the decrease. PMID- 15120721 TI - Respiratory tract infections: diagnosis and use of antibiotics by family physicians in north-eastern Poland. AB - The aim of our study was to describe the pattern of antibiotic prescribing for respiratory tract infections. Forty-four family physicians from Bialystok (north eastern Poland) filled in questionnaires regarding 1315 patients with respiratory tract infections. Antibiotics were prescribed in 803 cases (61.1%). The four principle diagnoses associated with prescribing an antibiotic were: bacterial pharyngitis/tonsillitis, acute bronchitis, unspecified bacterial superinfection and acute sinusitis for which a total of 572 antibiotic treatments were given (71.2% of all antibiotic prescriptions). The study shows that Polish family physicians diagnose respiratory tract infections empirically and frequently prescribe wide-spectrum antibiotics. More information is required about the diagnostic value of certain clinical symptoms such as purulent sputum, duration of cough and indications for broad-spectrum antibiotic use. PMID- 15120722 TI - Prevention of the selection of resistant Staphylococcus aureus by moxifloxacin plus doxycycline in an in vitro dynamic model: an additive effect of the combination. AB - Twenty-four hour ratios of area under the curve (AUC(24)) to MIC of 200-240 h providing quinolone concentrations above the mutant prevention concentration (MPC) protected from enrichment of resistant Staphylococcus aureus in our recent study that simulated the pharmacokinetics of moxifloxacin, gatifloxacin, levofloxacin and ciprofloxacin. These protective AUC(24)/MICs might also be achieved by using antibiotic combinations, assuming additive effects of two anti staphylococcal agents. To test this hypothesis, changes in S. aureus susceptibility were examined in a dynamic model that simulates 5-day treatment with moxifloxacin and doxycycline, alone and in combination at sub-optimal AUC(24)/MICs of each agent. Significant increases in MIC were observed with monotherapy where moxifloxacin or doxycycline concentrations fell into the mutant selection window (MSW) for more than 80% of the dosing interval (AUC(24)/MIC 60 h). Less pronounced changes in MIC occurred when the summed concentrations of moxifloxacin (AUC(24)/MIC 30 and 60 h) and doxycycline (AUC(24)/MIC 30 and 60 h) were inside the MSWs for the individual drugs for 30-50% of the dosing interval. No loss in susceptibility was found at moxifloxacin or doxycycline AUC(24)/MIC 170 h combined with the smaller AUC(24)/MIC (60 h) of the second compound. These data suggest that the total AUC(24)/MIC of 230 h might protect against S. aureus resistance. As this value is very close to that predicted in monotherapy with moxifloxacin (220 h), an additive protective effect of quinolone+doxycycline on the selection of resistant S. aureus is proposed. The use of drug combinations may be useful for restricting the enrichment of resistant mutants with agents whose clinically achievable AUC(24)/MICs do not provide concentrations above the MPC. PMID- 15120723 TI - Penicillin-induced killing and postantibiotic effect in oral streptococci are enhanced by platelet microbicidal proteins. AB - Thrombin-induced platelet microbicidal proteins (tPMP) are alpha-granule-derived cationic antimicrobial proteins released from platelets by stimulation with thrombin. tPMP has potent microbicidal activities against a broad spectrum of common microbial pathogens in infective endocarditis. We studied in vitro interactions of tPMP with penicillin against oral streptococci, Streptococcus rattus BHT (a tPMP-susceptible organism) and Streptococcus gordonii DL1 (a tPMP insusceptible organism). tPMP was prepared by stimulating rabbit platelets with thrombin. tPMP plus penicillin showed a synergistic bactericidal effect on both S. rattus BHT and S. gordonii DL1, in contrast to either agent alone. Sequential exposure of both S. rattus BHT and S. gordonii DL1 to tPMP followed by exposure to penicillin at 10x the MICs resulted in a significant extension of the postantibiotic-effect duration compared with antibiotic exposure alone. The combined data indicate that tPMP exerts cooperative bactericidal and growth inhibiting effect in concert with penicillin. PMID- 15120724 TI - Modulation of beta-lactam resistance in Staphylococcus aureus by catechins and gallates. AB - Aqueous extracts of Japanese green tea (Camellia sinensis) are able to reverse beta-lactam resistance in methicillin-resistant Staphylococcus aureus (MRSA). We have attributed the capacity to reverse oxacillin resistance in the homogeneous PBP2a producer BB568 and in EMRSA-16 to (-)-epicatechin gallate (ECG) and (-) catechin gallate (CG). Minimum inhibitory concentration (MIC) values for oxacillin were reduced from 256 and 512 to 1-4 mg/l, respectively, in the presence of these polyphenols. In addition, (-)-epigallocatechin gallate (EGCG) had a moderate capacity to modulate oxacillin activity against S. aureus BB568, but none against EMRSA-16. ECG, CG and EGCG increased the sensitivity of EMRSA-15 to oxacillin. The gallate moiety was essential for the oxacillin-modulating activity of ECG, as both (-)-epicatechin and (-)-epicatechin-3 cyclohexylcarboxylate were unable to reverse resistance to oxacillin. Gallic acid and three alkyl gallates (methyl gallate, propyl gallate, and octyl gallate) did not modulate beta-lactam resistance in MRSA. Octyl gallate exhibited direct antibacterial activity against S. aureus BB568 (16 mg/l). Modulation of beta lactam resistance by ECG significantly enhanced the activities of flucloxacillin and the carbapenem antibiotics imipenem and meropenem against 40 MRSA isolates, with MIC(90) values for the antibiotics reduced to the susceptibility breakpoint or below. Consequently, EGCG, CG and, particularly, ECG warrant further investigation as agents to combat beta-lactam resistance in S. aureus. PMID- 15120725 TI - Relative contribution of target gene mutation and efflux to fluoroquinolone and erythromycin resistance, in French poultry and pig isolates of Campylobacter coli. AB - Thirty-eight avian and swine French isolates of Campylobacter coli were studied for their mechanisms of co-resistance to fluoroquinolones and erythromycin. A Thr86Ile modification of GyrA, responsible for fluoroquinolone resistance, was found in all the strains. Two different levels of resistance to erythromycin (MIC of 8-16 or >/=256 mg/l) were observed. A A2075G mutation in the 23S rRNA genes was found only in the highly-resistant strains. Phe-Arg-beta-naphthylamide, an efflux pump inhibitor, potentiated erythromycin in all the strains examined but restored susceptibility only in the strains with a low-level of resistance. This suggests the involvement of efflux in intrinsic and in acquired low-level of resistance to erythromycin in C. coli. PMID- 15120726 TI - Contribution of AGC to ACC and other mutations at codon 315 of the katG gene in isoniazid-resistant Mycobacterium tuberculosis isolates from the Middle East. AB - The presence of ACC and other mutations at codon 315 in the katG gene was detected by PCR amplification followed by restriction fragment length polymorphism (PCR-RFLP) generated with restriction enzymes Msp I and MspA1 I in 37 isoniazid-resistant and 22-susceptible Mycobacterium tuberculosis isolates from Kuwait obtained in 2001. The mutation AGC to ACC was detected in 22 (60%) isolates while any mutation at codon 315 of the katG gene was present in 24 (65%) of 37 isoniazid-resistant isolates. The typing studies showed that majority of the isolates carrying mutations at codon 315 exhibited unique DNA banding patterns. The results were extended by additional analysis of 67, 28 and 17 isoniazid-resistant and 18, seven and six-susceptible M. tuberculosis isolates from Kuwait, Dubai and Beirut, respectively, that were analyzed previously for ACC mutation alone. These studies showed that one of 21, one of 10 and two of 11 isolates (all recovered from patients of Middle Eastern origin) with no AGC to ACC mutation from Kuwait, Dubai and Beirut, respectively, contained other mutations at codon 315 of the katG gene. None of the susceptible strains contained any mutation at codon 315. The PCR-RFLP with MspA1 I that detects all mutations at codon 315, compared with Msp I that detects only ACC mutation, identified more isoniazid-resistant strains with mutations at codon 315 in the katG gene. The data also showed that mutations other than AGC to ACC at codon 315 in the katG gene occur frequently in M. tuberculosis isolates recovered from Middle Eastern patients and should be incorporated in a rapid screen for the detection of mutations for isoniazid-resistance in the katG gene from this ethnic group. PMID- 15120727 TI - Antimicrobial susceptibility of clinical isolates of Enterobacteriaceae producing complex beta-lactamase patterns including extended-spectrum enzymes. AB - The antimicrobial susceptibility of 103 clinical isolates of Enterobacteriaceae to 11 antibiotics, was investigated, using a conventional inoculum size (5 x 10(5) CFU) and a higher inoculum size (5 x 10(8) CFU). All the isolates produced complex beta-lactamase patterns, including an extended-spectrum beta-lactamase (ESBL) of the TEM- or SHV-type plus other enzymes (a TEM-type or an SHV-type non ESBL and/or a class C enzyme). The following repertoire of ESBLs was produced by the isolates: TEM-15, TEM-19, TEM-26, TEM-52, TEM-72, TEM-87, TEM-92, SHV-2a, SHV 5 and SHV-12, as assessed by sequencing. Production of the other enzymes was showed by analytical isoelectric focusing. Overall, meropenem was the most active agent and less influenced by inoculum size, while other beta-lactams showed a lower activity and a significant inoculum size effect. In conclusion, from its in vitro performance, meropenem could be considered as the last resource drug against strains producing complex beta-lactamase patterns including an ESBL. PMID- 15120728 TI - Susceptibility of the Acinetobacter calcoaceticus-A. baumannii complex to imipenem, meropenem, sulbactam and colistin. AB - The Acinetobacter calcoaceticus-Acinetobacter baumannii complex includes some of the most clinically relevant species of the genus Acinetobacter due to their capacity to cause epidemic nosocomial outbreaks as well as their increasing resistance to antibiotics. Susceptibility of Acinetobacter strains varies greatly depending on origin, thus highlighting the importance of local analyses of susceptibility profiles. Two hundred twenty-one strains of the A. calcoaceticus A. baumannii complex were identified using biochemical tests and were biotyped. Strain susceptibility to imipenem, meropenem, colistin and sulbactam was studied using agar dilution. Eight different biotypes were found, type 1 accounting for 69.2% of the strains. MIC(50) and MIC(90) to imipenem, meropenem, colistin and sulbactam were 4 and 8 mg/l, 16 and 32 mg/l, 0.5 and 1mg/l, and 8 and 16 mg/l, with susceptibility rates of 64.3, 22.6, 98.2 and 73.8%, respectively. Biotype 1 was the most resistant. A statistically significant difference was observed for the mean MIC of the four predominant biotypes to imipenem, meropenem and sulbactam but not to colistin. PMID- 15120729 TI - Impact of tazobactam pharmacokinetics on the antimicrobial effect of piperacillin tazobactam combinations. AB - Pharmacokinetic-pharmacodynamic (PK-PD) modelling was used to study the impact of the pharmacokinetics of tazobactam on the antimicrobial effect of piperacillin tazobactam combinations. An in vitro experiment using a novel dilution system was performed to compare the effects of two conditions of the combination therapy against Escherichia coli ATCC35218, a beta-lactamase producing bacterium. Both conditions simulated the same initial concentrations of piperacillin and tazobactam, but different elimination half-lives for tazobactam. The killing and regrowth kinetics of E. coli clearly indicated that there is a difference in the antimicrobial effects when there is a difference in the pharmacokinetics of tazobactam in the combination therapy. The results show that for equal piperacillin exposure, different tazobactam half-lives will have a significant effect on antimicrobial outcome. PMID- 15120730 TI - Efficacy of clarithromycin against Streptococcus pneumoniae expressing mef(A) mediated resistance. AB - As a result of macrolide resistance rates of 25% for pneumococci in the US, the clinical use of this class as empirical therapy has been questioned. However, macrolides continue to be used with clinical success. Using an immunocompromised murine pneumonia model, this study evaluated in vivo efficacy of human simulated exposures of clarithromycin for 62 isolates of Streptococcus pneumoniae considered resistant by current methods of breakpoint determinations. Changes in bacterial density were compared between treated animals and untreated controls. Inhibition of bacterial growth was consistently observed for the majority of isolates tested with mean (S.D.) reductions in logCFU per lung of -0.88 (0.69), 1.02 (0.87), -0.47 (0.79), -0.84 (0.66), -0.25 (0.26), -0.80 (0.72) and -0.58 (0.47) for MICs of 1, 2, 4, 8, 16, 32 and 64 mg/l, respectively. A beneficial treatment effect was clearly noted for isolates with clarithromycin MICs <==8 mg/l. However, the sample size of isolates tested beyond the MIC of 8 mg/l was diminished due to mortality in both treated and untreated animals. Consistent suppression of bacterial growth observed in this neutropenic model provides support for the in vivo efficacy of clarithromycin with low-level macrolide resistant S. pneumoniae. PMID- 15120731 TI - Rapid detection and differentiation method of VanA, VanB and VanC phenotypes in vancomycin-resistant enterococci. AB - We developed a simple method that can replace the polymerase chain reaction (PCR) to distinguish between vancomycin-resistant enterococci (VRE) with the vanA, vanB and vanC genes. The method is based on induction of teicoplanin resistance by vancomycin in vanB-VRE, while the two compounds have a synergistic effect in vanC VRE. In addition, vanA-VRE shows resistance to both vancomycin and teicoplanin, and both the compounds can induce resistance to vanA-VRE. Utilising these properties, we attempted to develop a simple method to distinguish between vanA, vanB and vanC. We compared our simple method with the PCR method in 43 strains of vanA-VRE, 35 strains of vanB-VRE and 37 strains of vanC-VRE. The results were 100% consistent with that obtained by PCR. PMID- 15120732 TI - Resistance of uropathogens in symptomatic urinary tract infections in Leon, Nicaragua. AB - Management of urinary tract infections (UTI) in Central America and especially Nicaragua, is complicated by the lack of knowledge about the antibiotic resistance of uropathogens. We conducted a prevalence study to gain more insight into the aetiology, bacterial resistance and risk factors for symptomatic UTI in the region of Leon, Nicaragua. In 2002, all consecutive patients with UTI symptoms and pyuria >/=10 WBC/hpf were admitted to the study. Positive cultures from midstream urine specimens were defined as >/=10(5) cfu/ml of a single uropathogen. Susceptibility tests were performed with disc diffusion tests using the Kirby-Bauer method and broth microdilution using National Committee for Clinical Laboratory Standards criteria both in Leon and a reference laboratory in Utrecht. A positive culture was present in 62 of 208 study subjects (30%). Escherichia coli (56%), Klebsiella spp. (18%) and Enterobacter spp. (11%) were the most frequent pathogens isolated. Presence of cystocele, incontinence and increasing age were risk factors for bacterial UTI. E. coli was least resistant to ceftriaxone, amikacin and nitrofurantoin (>90% susceptible). We observed high resistance rates in E. coli to amoxicillin (82%, MIC(90) 128 mg/l), trimethoprim sulphamethoxazole (TMP-SMX) (64%, MIC(90) 32 mg/l), cephalothin (58%, MIC(90), 32 mg/l), ciprofloxacin (30%; MIC(90), 32 mg/l), amoxicillin/clavulanate (21%, MIC(90) 8 mg/l) and gentamicin (12%, MIC(90) 2 mg/l). Our results suggests that community acquired uropathogens in Nicaragua are highly resistant to many antimicrobial agents. The use of amoxicillin, trimethoprim-sulphamethoxazole and cephalothin against uropathogens needs to be reconsidered. High quinolone resistance rates among E. coli in Nicaragua gives cause for great concern. PMID- 15120733 TI - Linezolid and quinupristin/dalfopristin resistance in vancomycin-resistant enterococci and methicillin-resistant Staphylococcus aureus prior to clinical use in Turkey. AB - The aim of this study was to determine the in vitro antimicrobial activity of recently licensed quinupristin/dalfopristin and linezolid which have not yet been in clinical use in Turkey against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE) strains isolated from various clinical specimens by using the Etest. The results showed that all MRSA strains were fully susceptible to both the new compounds. All strains were inhibited by 1 mg/l quinupristin/dalfopristin (mode MIC 0.38 mg/l) and by 3 mg/l linezolid (mode MIC 1.5 mg/l). Four strains of Enterococcus faecium showed an increase of resistance of 2-3 mg/l to quinupristin/dalfopristin (susceptible mode MIC 0.38 mg/l). With linezolid, all strains except two fell within the range 0.75-2.0 mg/l. PMID- 15120735 TI - Susceptibility of Streptococcus pyogenes to two macrolides in northern Israel. AB - In the present study, the minimal inhibitory concentration (MIC) of azithromycin and roxithromycin for 200 Streptococcus pyogenes isolates from outpatients with tonsillopharyngitis were determined using Etest. All but one (99.5%) of the isolates were sensitive to both antibiotics; the MIC of the resistant isolate being 12 mg/l to azithromycin and 32 mg/l to roxithromycin. In this region, macrolides remain the drug of choice for the treatment of patients with S. pyogenes tonsillitis who present allergy to penicillin. The routine testing of susceptibility of S. pyogenes to macrolides in northern Israel is not justified. PMID- 15120734 TI - In vitro activity of mupirocin and amoxicillin-clavulanate alone and in combination against staphylococci including those resistant to methicillin. AB - Mupirocin and amoxicillin-clavulanate were synergistic against 9 of 49 (18%) strains of methicillin-resistant and methicillin-susceptible Staphylococcus aureus and coagulase-negative staphylococci (CNS). A pattern of enhanced killing was also found using time-kill studies. Time-kill assays were more discriminatory than chequerboard titration assays in demonstrating synergy. These results suggest that combinations of amoxicillin-clavulanate and mupirocin may have therapeutic benefits in prophylaxis against staphylococcal infections. PMID- 15120736 TI - Differential effects of the combination of caspofungin and terbinafine against Candida albicans, Candida dubliniensis and Candida kefyr. AB - The activity of caspofungin (CSP) combined with terbinafine (TRB) against Candida dubliniensis, Candida kefyr and azole-resistant Candida albicans was evaluated in vitro by checkerboard analysis. The combination of CSP with TRB resulted in positive interactive effects in vitro against C. albicans and C. kefyr but not against C. dubliniensis. Moreover, true synergism was observed only against TRB resistant strains which became susceptible to this drug in the presence of CSP. In contrast, indifference was observed against strains that were already sensitive to TRB indicating that CSP may inhibit resistance to TRB. PMID- 15120737 TI - Antiherpetic mode of action of (22S,23S)-3beta-bromo-5alpha,22,23 trihydroxystigmastan-6-one in vitro. AB - The replication of herpes simplex virus (HSV) type 1 in Vero cells is inhibited in the presence of (22S,23S)-3beta-bromo-5alpha,22,23-trihydroxystigmastan-6-one (6b), a synthetic brassinosteroid derivative. Attempts to disclose the mode of action of 6b indicate that a late step of virus multiplication is affected. In the presence of 6b, HSV late protein synthesis was severely diminished and this inhibitory effect of 6b on HSV antigen expression was confirmed by immunofluorescence assays. PMID- 15120738 TI - Drug resistance of Shigella species: changes over 20 years in Turkey. PMID- 15120739 TI - High incidence of vancomycin resistance in Enterococcus faecalis strains in a French hospital. PMID- 15120740 TI - Inhibition of multidrug transporters by verapamil or probenecid does not alter blood-brain barrier penetration of levetiracetam in rats. AB - Overexpression of multidrug efflux transporters such as P-glycoprotein (Pgp; ABCB1) or multidrug resistance proteins (MRPs; ABCC) in the blood-brain barrier has recently been suggested to explain, at least in part, pharmacoresistance in epilepsy, which affects about 30% of all patients with this common brain disorder. The novel antiepileptic drug (AED) levetiracetam (LEV) is an effective and well tolerated drug in many patients with otherwise AED-refractory epilepsy. One explanation for the favorable efficacy of LEV in pharmacoresistant patients would be that LEV is not a substrate for Pgp or MRPs in the BBB. In the present study, we used in vivo microdialysis in rats to study whether the concentration of LEV in the extracellular fluid of the cerebral cortex can be modulated by inhibition of Pgp or MRPs, using the Pgp inhibitor verapamil and the MRP1/2 inhibitor probenecid. Local perfusion with verapamil or probenecid via the microdialysis probe did not increase the extracellular brain concentration of LEV, which is in contrast to various other AEDs which have been studied previously by the same experimental protocol in this model. The data indicate that brain uptake of LEV is not affected by Pgp or MRP1/2 which may be an important reason for its antiepileptic efficacy in patients whose seizures are poorly controlled by other AEDs. PMID- 15120741 TI - Recurrent excitation in the dentate gyrus of a murine model of temporal lobe epilepsy. AB - Similar to rats, systemic pilocarpine injection causes status epilepticus (SE) and the eventual development of spontaneous seizures and mossy fiber sprouting in C57BL/6 and CD1 mice, but the physiological correlates of these events have not been identified in mice. Population responses in granule cells of the dentate gyrus were examined in transverse slices of the ventral hippocampus from pilocarpine-treated and untreated mice. In Mg(2+)-free bathing medium containing bicuculline, conditions designed to increase excitability in the slices, electrical stimulation of the hilus resulted in a single population spike in granule cells from control mice and pilocarpine-treated mice that did not experience SE. In SE survivors, similar stimulation resulted in a population spike followed, at a variable latency, by negative DC shifts and repetitive afterdischarges of 3-60 s duration, which were blocked by ionotropic glutamate receptor antagonists. Focal glutamate photostimulation of the granule cell layer at sites distant from the recording pipette resulted in population responses of 1 30 s duration in slices from SE survivors but not other groups. These data support the hypothesis that SE-induced mossy fiber sprouting and synaptic reorganization are relevant characteristics of seizure development in these murine strains, resembling rat models of human temporal lobe epilepsy. PMID- 15120742 TI - Prolonged changes in Ca2+/calmodulin-dependent protein kinase II after a brief pentylenetetrazol seizure; potential role in kindling. AB - This study evaluated the alteration of CaMKII autophosphorylation and distribution in rat brain following a single, brief pentylenetetrazol (PTZ) seizure and during PTZ kindling. Total CaMKII alpha subunit (alpha-CaMKII) and alpha-CaMKII phosphorylated at Thr(286) were detected by immunoblot. A large decrease in CaMKII Thr(286) phosphorylation, as well as CaMKII translocation from particulate to soluble fraction was observed in both cerebral cortex and hippocampus 0.5-4 h after the brief PTZ convulsion. These changes reverted to control values by 12 h. These long-lasting changes in CaMKII autophosphorylation and subcellular distribution after a brief seizure suggested that CaMKII could be involved in carrying forward the signal resulting from brief seizure activity, at least for a few hours, as would be required for kindling to occur. In PTZ kindled rats, convulsions produced changes in CaMKII Thr(286) phosphorylation and distribution in the same direction and of similar magnitude as after the acute convulsion, but lasting for a much longer time. In fact, reduced Thr(286) phosphorylation of alpha-CaMKII was observed up to 48 h, completely bridging the interval between PTZ injections. Similar, but intermediate changes were found in tissue from rats that were only partially kindled. These results implicate CaMKII as a molecular messenger in the acquisition of PTZ kindling. PMID- 15120743 TI - Effect of lamotrigine treatment on epileptogenesis: an experimental study in rat. AB - Prevention of epileptogenesis in patients with acute brain damaging insults like status epilepticus (SE) is a major challenge. We investigated whether lamotrigine (LTG) treatment started during SE is antiepileptogenic or disease-modifying. To mimic a clinical study design, LTG treatment (20 mg/kg) was started 2 h after the beginning of electrically induced SE in 14 rats and continued for 11 weeks (20 mg/kg per day for 2 weeks followed by 10 mg/kg per day for 9 weeks). One group of rats (n = 14) was treated with vehicle. Nine non-stimulated rats with vehicle treatment served as controls. Outcome measures were occurrence of epilepsy, severity of epilepsy, and histology (neuronal loss, mossy fiber sprouting). Clinical occurrence of seizures was assessed with 1-week continuous video electroencephalography monitoring during the 11th (i.e. during treatment) and 14th week (i.e. after drug wash-out) after SE. LTG reduced the number of electrographic seizures during SE to 43% of that in the vehicle group (P < 0.05). In the vehicle group, 93% (13/14), and in the LTG group, 100% (14/14) of the animals, developed epilepsy. In both groups, 64% of the rats had severe epilepsy (seizure frequency >1 per day). The mean frequency of spontaneous seizures, seizure duration, or behavioral severity of seizures did not differ between groups. The severity of hippocampal neuronal damage and density of mossy fiber sprouting were similar. In LTG-treated rats with severe epilepsy, however, the duration of seizures was shorter (34 versus 54s, P < 0.05) and the behavioral seizure score was milder (1.4 versus 3.4, P < 0.05) during LTG treatment than after drug wash-out. LTG treatment started during SE and continued for 11 weeks was not antiepileptogenic but did not worsen the outcome. These data, together with earlier studies of other antiepileptic drugs, suggest that strategies other than Na(+)-channel blockade should be explored to modulate the molecular cascades leading to epileptogenesis after SE. PMID- 15120744 TI - Up-regulation of the extracellular matrix glycoprotein tenascin-R during axonal reorganization and astrogliosis in the adult rat hippocampus. AB - Interactions between cells and extracellular matrix (ECM) molecules play a crucial role during brain development. The ECM glycoprotein tenascin-R (TN-R) has been implicated in the control of axon targeting, neural cell adhesion, migration and differentiation. Here, we have focused on the putative role of TN-R in chronic brain diseases involving increased neuronal excitability, as found in epilepsy. An episode of pilocarpine-induced status epilepticus (SE) led over a period of 3-30 days to neuron loss in the hippocampal hilus, CA3 and CA1 with reactive mossy fiber sprouting and astrogliosis in these regions. We found a focal up-regulation of granular TN-R immunoreactivity within the neuropil of segments of the CA3 pyramidal cell layer, the extent of this up-regulation paralleled the degree of pyramidal cell loss, mossy fiber sprouting and astrogliosis in these CA3 segments. In contrast, parvalbumin immunoreactivity and Wisteria floribundi agglutinin (WFA)-labeled perineuronal nets were reduced in CA3 segments with neuronal cell loss. The parallel development of increase in focal granular TN-R immunoreactivity, reactive mossy fiber sprouting and astrogliosis in CA3 implies a role for TN-R in axon targeting and synapse formation and/or in astrocytic targeting and interactions with the ECM during lesion-induced sprouting in the adult brain. PMID- 15120745 TI - The relation between extent of dorsal hippocampal kindling and delayed-match-to place performance in the Morris water maze. AB - Dorsal hippocampal kindling impairs subsequent performance on spatial tasks. The relation between this effect and the extent of kindling achieved prior to testing has not been clearly established. Thus, the present study investigated the effects of dorsal hippocampal kindling on performance of a delayed-match-to-place (DMTP) task in the Morris water maze by assessing performance after each of series of different points in the kindling process including 1, 6, 11, and 16 afterdischarges, 1 stage 1 seizure, and 1 stage 5 seizure. We found that kindling produced a deficit that was apparent very early into kindling in terms of both direct swim (by 1 AD) and escape distance (by 6 ADs) measures but that did not clearly change in severity with further kindling. These results illustrate that kindling of even a few localized hippocampal seizures can disrupt spatial cognition and suggest that the mechanisms mediating memory disruption either do not change substantially as kindling progresses or that compensatory processes are engaged across training that mitigate any further kindling-related deteriorations in performance. PMID- 15120746 TI - Behavioral effects of bicuculline microinjection in the dorsal versus ventral hippocampal formation of rats, and control of seizures by nigral muscimol. AB - This work aims to describe behavioral/electroencephalographic (EEG) seizures induced by bicuculline microinjection intracerebroventricularly (ICV) and in the dorsal hippocampal formation (DHF) or ventral hippocampal formation/amygdala area (VHF-AMY). We also test if GABAergic manipulation in the substantia nigra pars reticulata (SNPR) is capable of controlling those seizures. ICV injection of bicuculline induced a progressive sequence of convulsive responses, jumps and escapes from the open-field. This effect was partially reached by bicuculline injection in the DHF or VHF-AMY injection. Also: muscimol injection, but not GABA uptake blockers (nipecotic acid or a spider venom neurotoxin FrPbA2), into the SNPR abolished seizures induced by bicuculline injection in the DHF. It was concluded that different neuronal circuitry in the hippocampal formation are modulated, at least partially by nigral GABAergic mechanisms. PMID- 15120747 TI - Anticonvulsant effects of levetiracetam and levetiracetam-diazepam combinations in experimental status epilepticus. AB - Status epilepticus (SE) is a neurological emergency, with high mortality and high morbidity among survivors, and novel therapeutic agents are needed to improve this picture. We examined the effects of the antiepileptic drug levetiracetam (LEV) in an experimental model of self-sustaining status epilepticus (SSSE), induced in rats by electrical stimulation of the perforant path. LEV's unique spectrum of anticonvulsant activity, very high therapeutic index, and neuroprotective properties, make it a potentially interesting agent in the treatment of SE. Pretreatment with LEV intravenously reduced (30 mg/kg) or prevented (50-1000 mg/kg) the development of self-sustaining seizures. Treatment during the maintenance phase of SSSE diminished (at 200 mg/kg) or aborted seizures (in doses of 500 or 1000 mg/kg). Addition of LEV significantly enhanced the anticonvulsant effects of diazepam (DZP), even when both drugs where given in doses far below their therapeutic level. We conclude that LEV deserves further evaluation in the treatment of status epilepticus. PMID- 15120748 TI - Association between variation in the human KCNJ10 potassium ion channel gene and seizure susceptibility. AB - PURPOSE: Our research program uses genetic linkage and association analysis to identify human seizure sensitivity and resistance alleles. Quantitative trait loci mapping in mice led to identification of genetic variation in the potassium ion channel gene Kcnj10, implicating it as a putative seizure susceptibility gene. The purpose of this work was to translate these animal model data to a human genetic association study. METHODS: We used single stranded conformation polymorphism (SSCP) electrophoresis, DNA sequencing and database searching (NCBI) to identify variation in the human KCNJ10 gene. Restriction fragment length polymorphism (RFLP) analysis, SSCP and Pyrosequencing were used to genotype a single nucleotide polymorphism (SNP, dbSNP rs#1130183) in KCNJ10 in epilepsy patients (n = 407) and unrelated controls (n = 284). The epilepsy group was comprised of patients with refractory mesial temporal lobe epilepsy (n = 153), childhood absence (n = 84), juvenile myoclonic (n = 111) and idiopathic generalized epilepsy not otherwise specified (IGE-NOS, n = 59) and all were of European ancestry. RESULTS: SNP rs#1130183 (C > T) alters amino acid 271 (of 379) from an arginine to a cysteine (R271C). The C allele (Arg) is common with conversion to the T allele (Cys) occurring twice as often in controls compared to epilepsy patients. Contingency analysis documented a statistically significant association between seizure resistance and allele frequency, Mantel-Haenszel chi square = 5.65, d.f. = 1, P = 0.017, odds ratio 0.52, 95% CI 0.33-0.82. CONCLUSION: The T allele of SNP rs#1130183 is associated with seizure resistance when common forms of focal and generalized epilepsy are analyzed as a group. These data suggest that this missense variation in KCNJ10 (or a nearby variation) is related to general seizure susceptibility in humans. PMID- 15120749 TI - Influence of sympathetic autonomic arousal on cortical arousal: implications for a therapeutic behavioural intervention in epilepsy. AB - Negative amplitude shifts of cortical potential are related to seizure activity in epilepsy. Regulation of the cortical potential with biofeedback has been successfully used to reduce the frequency of some patients' seizures. Although such behavioural treatments are increasingly popular as an alternative to pharmacotherapy, there has been no investigation of the mechanisms that might bridge the behavioural index of peripheral autonomic activity and the central regulation of arousal. Galvanic Skin Response (GSR) is a sensitive measurement of autonomic arousal and physiological state which reflects one's behaviour. Thus we investigated the effect of peripheral autonomic modulation on cortical arousal with the future intention of using GSR biofeedback as a therapeutic treatment for epilepsy. The cortical negative potential was induced using the paradigm called Contingent Negative Variation (CNV) and measured in different physiological states. A high skin resistance state (reflecting a state of relaxation) and a low skin resistance state (reflecting a state of arousal), were engendered by two opposing procedures of GSR biofeedback. The CNV negative potential, acting as an index of cortical excitation, was significantly greater in amplitude at high levels of skin resistance (relaxed state) than at low levels of skin resistance (aroused state). Our results suggest an inverse relationship between a peripheral measure of autonomic arousal and an index of cortical arousal, the CNV. Moreover, we demonstrate modulation of this arousal-related potential by a behavioural intervention, indicating a potential therapeutic use of arousal biofeedback using GSR in the management of treatment-resistant epilepsy. PMID- 15120751 TI - Circadian rhythms on hypothalamic-pituitary-adrenal axis hormones and cytokines of collagen induced arthritis in rats. AB - This study is designed to identify whether circadian rhythms of the hormones of the hypothalamic-pituitary-adrenal (HPA) axis are associated with corresponding circadian fluctuations in cytokines in a rat model of collagen-induced arthritis (CIA). CIA is induced in Wistar rats by an intradermal injection of bovine type II collagen emulsified with complete adjuvant at the left foot. On day 33, in both the CIA and the control rats, circulating adrenocorticotropin hormone (ACTH) and corticosterone, tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and IL-1beta were evaluated at 6 h intervals from 00:00 to 24:00, and analyzed by statistics and cosinor-rhythmometry. The results showed that plasma corticosterone in CIA rats had a trough at 18:00 and reached a peak at 06:00 significantly. While peak values were presented in TNF-alpha at 24:00 and in IL-6 from 06:00 and 18:00 to 24:00. CIA rats exhibit abnormal circadian rhythms, with degrading amplitudes of corticosterone and IL-6, elevating amplitude of TNF alpha, and marked phase shifts in corticosterone and IL-6. Our investigation suggests that the disorders of HPA axis in CIA rats may be related to the influence of inflammation mediators on hypothalamic centers. The circadian rhythms of hormones and cytokines in CIA rats may be reset due to the defective function of the HPA axis. PMID- 15120750 TI - Immunosuppressive and anti-inflammatory action of antioxidants in rat autoimmune diabetes. AB - Oxidative stress makes an important contribution to the development of autoimmune diabetes. We therefore tested the possible therapeutic value of two anti oxidants, butylated hydroxyanisole (BHA) and pyrrolidine dithiocarbamate (PDTC), in the animal model of diabetes induced in susceptible DA rats by multiple low doses of streptozotocin (MLD-SZ, 20 mg/kg/day for 5 days). Administration of either BHA, or PDTC (50 mg/kg/day for 7 days), after finishing MLD-SZ injections, attenuated both the development of hyperglycemia and insulitis. Ex vivo analysis revealed that BHA treatment reduced the proliferation of autoreactive lymphocytes and down-regulated their adhesion to endothelium. In addition, BHA markedly attenuated the production of proinflammatory cytokines IL-1beta and TNF-alpha by both islets of pancreas and peritoneal macrophages. In parallel, macrophage release of cytotoxic oxygen and nitrogen intermediates superoxide anion (O(2)*( )) and nitric oxide (NO*), respectively, was significantly inhibited. Finally, BHA treatment reduced intrapancreatic expression of inducible NO synthase (iNOS) and consequent production of NO* by pancreatic islets. Together, these data indicate that antioxidant agents might be a feasible therapeutic tools to interfere with development of autoimmune diabetes at multiple levels, including lymphocyte proliferation and adhesion, as well as the production of proinflammatory and cytotoxic mediators. PMID- 15120752 TI - Transplacental exposure to bafilomycin disrupts pancreatic islet organogenesis and accelerates diabetes onset in NOD mice. AB - Bafilomycin, a plecomacrolide produced by plant-pathogenic Streptomyces, contaminates tuberous vegetables and has adverse effects on beta cells in adult mice. We therefore determined whether dietary bafilomycin influenced the progression of diabetes in the non-obese diabetic (NOD) mouse model of autoimmune Type 1 diabetes. Parent NOD mice were fed sub-toxic doses of bafilomycin in drinking water from conception until weaning, or various times after birth and blood glucose was monitored in the offspring. Pancreatic islets in neonatal offspring were examined histologically by quantitative morphometry and islet cell apoptosis was estimated by TUNEL assay. Exposure in utero to bafilomycin but not after birth significantly accelerated onset and increased the frequency of diabetes. In exposed mice, pancreatic islet organogenesis was disrupted, characterized by a striking increase in beta-cell mass and a shift in timing of the normal wave of neonatal islet cell apoptosis from 2 weeks to 4 weeks of age. We postulate that accelerated onset and increased incidence of diabetes later in life result from disruption of the normal turnover of beta cells in the neonatal pancreas. Since bafilomycin and related plecomacrolides contaminate Streptomyces infected vegetables, dietary exposure during pregnancy could be an important and previously unsuspected environmental component of human Type 1 diabetes. PMID- 15120753 TI - Knock-out of the histidine decarboxylase gene modifies the repertoire of natural autoantibodies. AB - Natural antibodies (NA) are antibodies produced in the absence of known immunization with specific antigens. NA are found in the blood of healthy humans and mice. Histamine influences many aspects of the immune response, including antibody production. However, the role of histamine in the generation of NA has not yet been studied. In this work, we used an ELISA assay to characterize the self-antigen binding repertoires of NA in wild type (WT) mice and in histidine decarboxylase knock-out (HDC-KO) mice, unable to synthesize histamine. We now report that HDC-KO and WT mice differed in the patterns of autoreactivity of their IgM and IgG NA. The NA in HDC-KO sera manifested a larger repertoire of IgM autoantibodies than did the WT sera. The self-antigens bound by IgM from HDC-KO mice included structural proteins, enzymes associated with cellular metabolism, double-stranded and single-stranded DNA, and tissue-specific antigens like insulin. There were relatively fewer differences in the NA repertoire of IgG autoantibodies of the mice: notably, the HDC-KO sera reacted with glutamic acid decarboxylase (GAD), an antigen associated with autoimmune diabetes. These results demonstrate that endogenous histamine can influence the self-reactivity of the NA repertoire. PMID- 15120754 TI - Genetic differentiation of poly I:C from B:9-23 peptide induced experimental autoimmune diabetes. AB - Type 1 diabetes is an immune-mediated disease, in which T cells of the adaptive immune system mediate beta cell destruction. Recently the innate immune system has been linked to etiopathogenesis of several autoimmune diseases including type 1 diabetes, as innate effector cells (e.g. dendritic cells, monocytes/macrophages and NK cells) can prime and promote or regulate (auto)immune responses. We have previously developed an experimental autoimmune diabetes (EAD) model with insulin peptide B:9-23 immunization in transgenic H-2(d)mice expressing the costimulatory molecule B7.1 in their islets (under the Rat Insulin Promotor, RIP). We compared the induction of diabetes with polyinosinic-polycytidylic acid (Poly I:C), a mimic of double stranded viral RNA versus insulin B:9-23 peptide in mice following backcrossing of the B7.1 transgene on to BALB/c mice from original B7.1 C57Bl/6 mice. We find that diabetes induction by Poly I:C is C57Bl/6 associated, whereas B:9-23 peptide induced diabetes and induction of insulin autoantibodies (IAA) are dependent on BALB/c genes. This B:9-23 peptide induced diabetes is consistent with MHC class II H-2(d)being necessary for the response to this peptide. Of note Poly I:C induction of diabetes was lost while B:9-23 induction was retained with backcrossing to BALB/c mice. Interaction of genes and environment (antigenic epitope and viral mimic) can be important in the pathogenesis of immune mediated diabetes and activation of the innate immune system (e.g. Poly I:C) may be one key determinant. PMID- 15120756 TI - Autoantibodies in SLE but not in scleroderma react with protein-stripped nucleosomes. AB - Autoantibodies against nucleosomes (ANuA) are known to be sensitive markers for systemic lupus erythematosus (SLE), but their clinical relevance seemed to be limited because sera from patients with progressive systemic sclerosis (PSS) also showed positive reactions with conventional ANuA ELISA test systems (anti-Nu1 ELISA). It was generally assumed thatANuA were associated with both diseases. Using discontinuous sucrose gradient centrifugation to generate pure nucleosomes, we discovered by chance that at the 30-50% sucrose interface an antigen (Nu2) banded which was demonstrably free of non-histone components and histone H1. The two different nucleosome preparations, Nu1 and Nu2, were used in parallel as antigenic substrates in standardised ELISA tests to analyse sera from SLE (295 patients), PSS (119) and patients with other rheumatic diseases (101). With Nu1, 62% of the SLE and 52% of the PSS sera showed positive reactions. Two sera from patients suffering from Sjogren's syndrome (SS) and one from polymyositis were also positive. Using the Nu2 preparation, 58% of the SLE but none of the PSS sera showed a positive reaction. One serum from a patient with SS was also positive. It could be shown that it was the PSS-specific autoantigen Scl-70 in the nucleosome preparation (Nu1) which contributed to the positive reactions of the PSS sera in conventional ANuA test systems, whereas in the Nu2 preparation no remaining Scl-70 was detectable. The present study definitely proved that ANuA are highly and specifically associated with SLE but not with PSS. PMID- 15120755 TI - Reduced anti-TNFalpha autoantibody levels coincide with flare in systemic lupus erythematosus. AB - Deviating cytokine patterns, as a consequence of aberrant immunoregulation, is implicated to be of aetiopathogenetic importance in systemic lupus erythematosus (SLE). To evaluate the possibility of anti-cytokine autoantibody-mediated cytokine regulation/dysregulation, IgG class autoantibodies against cytokines (IL 1beta, IL-6, IL-10, TNFalpha and TGFbeta(1)) were analysed by enzyme-linked immunosorbent assay (ELISA) in serial serum samples from clinically well characterized SLE patients and in normal human sera (NHS). Anti-TNFalpha autoantibody levels were lower in patients with active disease compared to inactive disease (P<0.001) as well as to NHS (P<0.001). The anti-TNFalpha antibody levels correlated inversely to the SLE disease activity index (SLEDAI) (r(2)=0.07, P<0.01), whereas anti-TGFbeta antibodies were raised in SLE and correlated positively to levels of complement factor C1q (r(2)=0.08, P<0.005). Generally raised anti-cytokine antibody levels and correlations to disease activity measures were found in one individual. Inverse correlations were found comparing SLEDAI scores and autoantibodies to TNFalpha (r(2)=0.92) and IL-6 (r(2)=0.86) and positive correlations were found between levels of anti-TNFalpha and C1q (r(2)=0.86) and C3 (r(2)=0.90). We show, for the first time, a coincidence between reduced anti-TNFalpha autoantibody levels and disease exacerbation in SLE, which is of interest regarding aetiopathogenesis and disease control. PMID- 15120757 TI - Intrathecal synthesis of autoantibodies against tissue transglutaminase. AB - Anti-tissue transglutaminase (tTG) antibodies (AtTGA) are typically found in serum of patients with untreated coeliac disease (CD). tTG catalyses crosslinking of peptides an activity supposed to be important in neurological disorders. tTG occurs in cerebrospinal fluid (CSF) and its assay in CSF was suggested to be diagnostically useful. However, nothing is known about AtTGA in CSF. Therefore, in 129 unselected CSF-serum pairs IgA- and IgG-AtTGA were assayed by ELISA using human recombinant tTG. For comparison, IgA- and IgG-anti-gliadin antibodies (AGA), typically coexisting with AtTGA were measured. Albumin, total IgA and IgG and further parameters were determined according to routine programme recommended by the European CSF consensus group. AtTGA were detected in 27 (IgA) and in 63 (IgG) CSF samples. Antibody indices (AI) could be calculated for AtTGA from 21 (IgA) and from 61 (IgG) sample pairs. AI for AtTGA was >2 in 11 (IgA) and in 22 (IgG) sample pairs, hinting to intrathecal antibody synthesis. AI for AGA was >2 only for 1 (IgA) and 2 (IgG) sample pairs. Patients with normal routine findings had significantly higher AI for IgA-AtTGA than patients with abnormal findings. This is the first demonstration of AtTGA in CSF and their intrathecal synthesis. The pathogenetic relevance of this new autoantibody species remains to be clarified. PMID- 15120758 TI - Increased incidence of autoimmune disorders as a late complication in children with early onset dermatitis and/or milk allergy. AB - Subjects with atopic dermatitis and autoimmune disorders share some similar immune response disorders. The aim of this study was to see whether subjects with early onset atopic dermatitis run a risk of eventually developing autoimmune diseases. The results of a questionnaire of 145 adolescents (70 f, 75 m, mean age 18.2 years, range 16-23 years) was compared with those of a group of 262 controls (112 f, 150 m, mean age 17.5 years, range 16-21 years), 164 of whom reported no atopic symptoms and were treated as a separate group for statistical analysis. As compared with the non-atopic controls, the study group subjects showed a significantly increased incidence of autoimmune disorders (9% vs. 1%), the relative risk ratio of a subject with infantile onset atopic eczema getting a gastrointestinal (GI) immune-mediated disease being 2.4 (CI(95)2.1-2.8) and of getting some other autoimmune disorder 3.1 (CI(95)2.8-9.7). The positive skin prick tests showed a negative association with the manifestation of a GI or other autoimmune disorder. The subjects with infantile dermatitis also reported recurrent abdominal pains (23% vs. 15%), and milk-induced gastrointestinal symptoms (19% vs. 10%) significantly more even as young adults than the controls. Our study showed that infantile atopy increases a predisposition to autoimmune disorders, suggesting that these two entities might have a common immunological determinant. While a high incidence of chronic GI complaints among the study subjects suggests the ongoing activity of local immune responses. However, more detailed prospective studies are needed to confirm these observations. PMID- 15120759 TI - Effect of gender on T-cell proliferative responses to myelin proteolipid protein antigens in patients with multiple sclerosis and controls. AB - Multiple sclerosis (MS) is an inflammatory demyelinating disorder of the central nervous system. Gender influences both susceptibility to MS, with the disease being more common in women, and the clinical course of disease, with an increased proportion of males developing the primary progressive form of the disease. The basis for these differences may include genetic and immunological factors, and the immunological differences between men and women may be influenced by the effects of the sex hormones. Over several years we have collected blood from MS patients and controls, and measured T-cell responses to myelin proteolipid protein (PLP) and myelin basic protein (MBP) and have shown increased responses to PLP in MS patients compared to healthy controls and patients with other neurological diseases. In the present study we analyzed data from over 500 individuals, to determine whether there are differences between males and females in their responses to PLP and MBP. We found that there was higher frequency of increased T-cell reactivity to immunodominant PLP peptides in women than in men, particularly in non-MS individuals. We suggest that this may be relevant to the higher prevalence of MS in women. PMID- 15120760 TI - Disease-specific cross-reactivity between mimicking peptides of heat shock protein of Mycobacterium gordonae and dominant epitope of E2 subunit of pyruvate dehydrogenase is common in Spanish but not British patients with primary biliary cirrhosis. AB - Previous studies on Spanish patients with Primary Biliary Cirrhosis (PBC) have shown extensive, disease-specific cross-reactivity between the 65-kDa heat shock protein (hsp65) of Mycobacterium gordonae and pyruvate dehydrogenase complex-E2 (PDC-E2), the major target of anti-mitochondrial antibody (AMA). Studies on a British population were unable to substantiate these findings. Having found that there is an excellent and almost unique match between the PDC-E2 autoepitope and a sequence in mycobacterial hsp65s, we tested the corresponding peptides by ELISA for cross-reactivity using sera from 90 PBC patients, 40 Spanish and 50 British, and 84 pathological controls. Reactivity to the MYCGO hsp65(90-104)/human PDC E2(212-226)pair was present in 19 (47.5%) Spanish PBC patients and in 2 (4%) of the 50 British. Reactivity was not seen in any of the controls. Simultaneous reactivity to mimics was due to cross-reactivity as confirmed by inhibition studies. Three dimensional modelling predicts mycobacterial hsp65(90-104)to be exposed on the surface of the protein. The affinity of anti-hsp65(90-104)antibody was higher than that of anti-PDC-E2(212-226). Hsp65(90-104)is a target of disease specific cross-reactivity to PDC-E2(212-226). The geographical confinement of this phenomenon is probably the result of complex genetic, environmental and immunological interaction. PMID- 15120761 TI - Severe withdrawal from short-term transdermal fentanyl after naloxone for uremic pruritus. PMID- 15120762 TI - Chlorpheniramine maleate as an alternative to antiemetic cyclizine. PMID- 15120763 TI - Methadone mouthwash for the management of oral ulcer pain. PMID- 15120764 TI - Opioid consumption in Italy: industry is not necessarily business--look at the effects. PMID- 15120765 TI - Re: Anemia at the end of life. PMID- 15120767 TI - Re: Chronic lymphocytic leukemia resembling metastatic bone disease. PMID- 15120768 TI - Evaluation of a short-term group intervention for informal carers of patients attending a home palliative care service. AB - Despite evidence of high psychological distress and unmet needs, evaluated interventions for informal caregivers in palliative care are few. This study involved an observational outcome evaluation of attendees, and a comparison group, in specialist home palliative care. The measures included carer psychological status and patient physical status at baseline, 8 weeks, and 20 weeks. Qualitative data were collected regarding content, satisfaction with, and impact of intervention. Process data described the uptake, resources, and group activity. The intervention combined informal multiprofessional teaching with facilitated peer exchange and support, and was delivered over 6 sessions of 90 minutes per week. The uptake rate was 25%; carers were less likely to accept if they were in paid employment (OR=0.26, P=0.06), and more likely to accept if they utilized avoidance coping (OR=1.13, P=0.04) or their patient had worse physical status (OR=2.1, P=0.03). Attendees described significant support and knowledge gains from the multiprofessional input and peer group. Most relied on social comparison processes to appraise their situation. Potential detection of significant effects on global psychological scores (i.e. anxiety, depression, and burden) using multivariate analysis was disallowed due to attrition. This acceptable and accessible intervention provided information and support; further outcome studies are needed for a range of interventions. Short-term interventions are unlikely to affect global psychological scores, and future evaluations should include additional time points of data collection to demonstrate support during attendance. PMID- 15120769 TI - Use of strong opioids in advanced cancer pain: a randomized trial. AB - The World Health Organization (WHO) guidelines for the treatment of cancer pain recommend nonopioid analgesics as first-line therapy, so-called "weak" analgesics combined with nonopioid analgesics as second-line therapy, and so-called "strong" opioids (with nonopioid analgesics) only as third-line therapy. However, these guidelines can be questioned with regard to the extent of efficacy as well as the rationale for not using strong opioids as first-line treatment, especially in terminal cancer patients. The purpose of this randomized study was to prospectively compare the efficacy and tolerability of strong opioids as first line agents with the recommendations of the WHO in terminal cancer patients. One hundred patients with mild-moderate pain were randomized to treatment according to WHO guidelines or to treatment with strong opioids. Evaluated outcomes included pain intensity, need for change in therapy, quality of life, Karnofsky Performance Status, general condition of the patient, and adverse events. No between-treatment differences were observed for changes in quality of life or performance status, but patients started on strong opioids had significantly better pain relief than patients treated according to WHO guidelines (P=0.041). Additionally, patients started on strong opioids required significantly fewer changes in therapy, had greater reduction in pain when a change was initiated, and reported greater satisfaction with treatment than the comparator group (P=0.041). Strong opioids were safe and well-tolerated, with no development of tolerance or serious adverse events. These data suggest the utility of strong opioids for first-line treatment of pain in patients with terminal cancer. PMID- 15120770 TI - Cancer patient compliance in the self-administration of a pain assessment tool. AB - Accurate pain assessment is considered essential for effective management of cancer pain. The aim of this study was to evaluate the compliance of hospitalized patients with chronic cancer pain, referred to an inpatient palliative care consultation service, with self-assessment of pain intensity by means of a daily pain form. The form was distributed daily by the pain consult nurse and required three daily pain intensity measurements on 0 to 10 numerical scales, separately for pain at rest and pain on movement. Of 174 consecutive patients, 106 (61%) participated in the study and were followed up for a median of 10.6 days (range 1 32 days). Compliance was defined as the number of assessment forms completed over the number of evaluation days available for each patient. Mean compliance was 58%. The main reasons for not completing the form were related to subjective psychological variables (44%), physical distress (26%), and absence of pain (16%). Lack of understanding of the method was reported as the main reason for non-compliance by only 1% of patients. PMID- 15120771 TI - Relationships of mood disturbance and social support to symptom experience in Korean women with breast cancer. AB - The purpose of this study was to identify how mood disturbance and social support were related to the symptoms experienced by Korean women with breast cancer. A cross-sectional, correlational design was used for the study. A convenience sample of 134 Korean women receiving chemotherapy for breast cancer was recruited. The participants completed questionnaires on symptom experience using the Symptom Experience Scale, mood disturbance using the Linear Analogue Self Assessment Scale, and social support using the Social Support Scale. Mood disturbance and social support had a significant interaction effect on symptom experience. A higher level of mood disturbance led to a higher level of symptoms when the level of social support was average or low, which implies that clinical interventions for attenuating the impact of mood disturbance on symptom experience might be effective only for women perceiving average or low levels of social support. PMID- 15120772 TI - The bioavailability of morphine applied topically to cutaneous ulcers. AB - A number of studies have reported the analgesic effect of morphine when applied topically to painful skin ulcers. It has been suggested that morphine may exert a local action, as opioid receptors have been demonstrated on peripheral nerve terminals. In this study, we investigated the bioavailability of topically applied morphine to cutaneous ulcers. Six hospice inpatients with skin ulcers were given morphine sulfate 10 mg in Intrasite gel topically and morphine sulfate 10 mg subcutaneously over 4 hours, at least 48 hours apart, in randomized order. Morphine, morphine-6-glucuronide (M6G), and morphine-3-glucuronide (M3G) were determined in plasma using a specific HPLC method. In five patients morphine and its metabolites were undetectable when applied topically. In one patient (with the largest ulcer), morphine and M6G were detected. The calculated morphine and M6G bioavailability in this patient were 20% and 21%, respectively. M3G was also detected but was below the lower limit of quantitation. When applied topically to ulcers, morphine was not absorbed in the majority of patients, suggesting any analgesic effect would be mediated locally rather than systemically. However, in ulcers with a large surface area, systemic absorption may occur. PMID- 15120773 TI - Development of a self-report screening instrument for assessing potential opioid medication misuse in chronic pain patients. AB - This study constituted the first step in the psychometric development of a self report screening instrument for risk of opioid medication misuse among chronic pain patients. A 26-item instrument, the Pain Medication Questionnaire (PMQ), was constructed based on suspected behavioral correlates of opioid medication misuse, which heretofore have received limited empirical investigation. The PMQ was administered to 184 patients at an interdisciplinary pain treatment center. Reliability coefficients for the PMQ were found to be of moderate but acceptable strength. Construct and concurrent validity were examined through correlation of PMQ scores to measures of substance abuse, physical and psychological functioning, and physicians' risk assessments. To explore high and low cutoff points for misuse risk, subgroups were formed according to the upper and lower thirds of PMQ scores and compared on validity measures. Higher PMQ scores were associated with history of substance abuse, higher levels of psychosocial distress, and poorer functioning. Future psychometric analyses will consider predictive validity and examine shortened versions of the instrument. PMID- 15120774 TI - Valdecoxib versus rofecoxib in acute postsurgical pain: results of a randomized controlled trial. AB - The analgesic efficacy of the cyclooxygenase-2 specific inhibitors, valdecoxib and rofecoxib, were evaluated in patients following oral surgery. In a randomized, double-blind, controlled trial, patients experiencing moderate or severe pain received single-dose valdecoxib 40 mg (n=99), rofecoxib 50 mg (n=101), or placebo (n=50) within 4 hours after multiple third molar extraction with bone removal. Onset of action was significantly faster with valdecoxib 40 mg (30 minutes) compared with rofecoxib 50 mg (45 minutes), as measured by pain intensity difference and pain relief scores (P isoflurane > sevoflurane. Many reports suggest luxury perfusion at high concentrations of desflurane. Sevoflurane maintains intact cerebral autoregulation up to 1.5 MAC. Desflurane induces a significant impairment in autoregulation, with a completely abolished autoregulation at 1.5 MAC. Both sevoflurane and desflurane (up to 1.5 MAC) maintain normal CO(2) regulation. As to their effect on final intracranial pressure (ICP), both sevoflurane and desflurane revealed no increases in ICP. However, compared to intravenous hypnotics, subdural ICP is higher with volatiles because of their tendency to increase cerebral swelling after dura opening (isoflurane > sevoflurane). Several case reports have noted seizure-like movements, as well as EEG recorded seizures during induction of sevoflurane anesthesia. Especially, in children during inhalational induction with hyperventilation at a high sevoflurane concentration, severe epileptiform EEG with a hyperdynamic response were observed, which urges for caution using inhalational sevoflurane induction in children for neurosurgical procedures. Neuroprotective properties (reduced neuronal death either by necrosis or apoptosis) have been attributed to all volatile agents. However, these neuroprotective effects have been described in experimental or animal models, so their possible effect on humans remains to be proven. PMID- 15120784 TI - [Target-controlled infusion with propofol for neuro-anesthesia]. AB - Propofol is an intravenous anaesthetic agent, which presents interesting features for its use in neuro-anaesthesia: it is a powerful hypnotic that does not increase the intracranial pressure. The delay of recovery is short even after several hours of continuous infusion. This is essential for a fast neurologic examination. Continuous infusion should be preferred to bolus in order to prevent hypotension and decrease of the cerebral perfusion pressure. Target-controlled infusion models based on effect site concentrations are now available through several softwares. This technique appears especially useful for awake craniotomy and functional neurosurgery. The level of consciousness is easily fixed between deep anaesthesia and light sedation permitting to ask the patient to move following orders. A sedation controlled by the patient himself is even possible. PMID- 15120785 TI - [Opioid anesthetics (sufentanil and remifentanil) in neuro-anaesthesia]. AB - Remifentanil is the latest available compound of the 4-anilidopiperidine derivatives. It is characterized by an ultrashort duration of action and a metabolism independent of both hepatic and renal functions. Its main drawback is a lack of residual analgesia and the risk of postoperative hyperalgesia. Since its introduction in clinical practice, this drug has been compared to other congeners in a few studies in the setting of neurosurgery. Cerebral hemodynamics, intracranial pressure and CO(2) reactivity are similar to the effects of fentanyl and sufentanil provided that systemic arterial pressure is maintained. Haemodynamics does not greatly vary according to the type of the opioid. Fentanyl and sufentanil require higher hypnotic dosage (halogenated agents, propofol) and remifentanil is accompanied by greater volumes of fluid infusion. A marked reduction in extubation times and superior level of consciousness are reported with remifentanil. Rescue analgesic has to be given faster but pain scores remain low. Morphine 0.08 mg/kg IV administered at bone replacement results in good postoperative analgesia without delayed recovery. In summary, remifentanil is appropriate when rapid recovery and neurological evaluation are desired. Conversely, sufentanil is more suitable and easier to administer when postoperative mechanical ventilation and postponed awakening are scheduled. PMID- 15120786 TI - [Anaesthetic management of awake craniotomy]. AB - This review article presents a detailed analysis of patients' management for awake craniotomy, at the light of the available data in the literature and the authors' experience. Indications of this type of surgery are discussed as well as anaesthetic management itself, from preoperative assessment of the patient to peroperative concerns. Anaesthetic strategy, choice of anaesthetic agents, anaesthetic technique, and management of the airway and possible complications are discussed. The authors emphasize the tricky aspect of the procedure, the necessity of rigorous patient selection and good preparation. They emphasize the need for controlled studies to validate the proposed techniques. PMID- 15120787 TI - [The neuro-anaesthesiology assisted by the electroencephalogram]. AB - The electrophysiology can be considered as one of the most reliable assessment of the nervous system function during anaesthesia. For instance, the electroencephalogram (EEG) can be relatively easily simplified and adapted to the current practice of the neuro-anaesthesia or neuro-intensive medicine. Since approximately 10 years, some extremely simplified devices have been introduced, but they allowed only inaccurate assessment because of their too global analysis of the brain function. The monitoring of the true digitised EEG, based on a simplified brain mapping montage combined to the analysis in the frequency and the time domains corresponds to a new investigation tool: the simplified EEG or sEEG, which can be included in our usual monitors or anaesthesia and intensive care machines. This new sort of monitoring could give us valuable information about, first, the control of the peroperative consciousness and the nociceptive or not afferent inputs due to surgical stimulations, and, second, the detection of several particular physiopathologic states during neuro-anaesthesia (brain ischaemia, non-convulsive epilepsy, cerebral protection em leader ). These different theoretical and practical topics are reviewed to try to define the possible preliminary rules of the use of the sEEG. PMID- 15120789 TI - [Cerebral and systemic haemodynamic changes during neurosurgical recovery]. AB - Major complications after intracranial surgery occur in 13-27% of patients. Among multiple causes, haemodynamic and metabolic changes of anaesthesia recovery may be responsible for intracranial complications. Recovery from neurosurgical anaesthesia is followed by an increase in body oxygen consumption and catecholamines concentrations. However, in normothermic patients, theses changes are usually mild and not prevented by a 2-h recovery delay. Systemic hypertension is common after neurosurgery and a link between perioperative hypertension and intracranial haemorrhage has been established. The cerebral consequences of recovery associate cerebral hyperaemia and increased ICP in patients with a tight brain at the end of surgery. Cerebral hyperaemia may promote or exacerbate cerebral haemorrhage or oedema. This has been demonstrated in patients operated for subdural haematoma removal or undergoing carotid surgery. Prevention of hypothermia and pain are key factors to prevent metabolic changes. Beta-blockers seem to be suitable agents to obtain haemodynamic control in neurosurgical patients. PMID- 15120788 TI - [Management of neurosurgical patient operated upon for intracranial tumour]. AB - 1. Neurological state of patient. PROCEDURES: (low risk of ICP problems or ischemia, little need for brain relaxation). - Volatile-based technique; "high risk" procedures (anticipated ICP problems, significant risk of intraoperative cerebral ischemia, need for excellent brain relaxation): use total intravenous anaesthesia. EXTRACRANIAL MONITORING: For example, cardiovascular or renal, venous air embolism. Intracranial monitoring. - General environment vs. specific functions-metabolic (jugular venous bulb), neurophysiological (EEG/EP), functional (transcranial Doppler). 4. Induction of anaesthesia. GOALS: Ventilatory control (early mild hyperventilation; avoid hypercapnia, hypoxemia); blood pressure control (avoid CNS arousal: adequate antinociception, anaesthesia); optimal position on ICP-volume curve. PATIENT POSITIONING: Pin holder application --> maximal nociceptive stimulus, block by deeper anaesthesia or analgesia and local anesthetic pin site infiltration. Alternative: antihypertensives. 5. Maintenance of anaesthesia. GOALS: Controlling brain tension via control of CMR and CBF: preventing CNS arousal (depth of anaesthesia, antinociception); treating consequences of CNS arousal (sympatholysis, antihypertensives); the "chemical brain retractor concept". NEUROPROTECTION: Maintenance of an optimal intracranial environment (matching cerebral substrate demand and supply). 6. Emergence from anaesthesia. GOALS: Maintain intra/extracranial homeostasis. Avoid factors --> intracranial bleeding and/or increasing CBF/ICP. The patient should be calm, co-operative and responsive to verbal commands soon after emergence. EARLY VS. LATE EMERGENCE: Ideal: rapid emergence to permit early assessment of surgical results and postoperative neurological follow-up, but there are still some categories of patients where early emergence is not appropriate. PMID- 15120790 TI - [Pain and postoperative analgesia after craniotomy]. AB - Neurosurgery has for a long time been considered as a minimal painful surgery. This explains why there are few references in the literature concerning postoperative neurosurgical pain. Recent papers have demonstrated that even if postoperative pain is less important than in other specialities, such pain exists and should be taken care of. Rapid neurological recovery is now possible because of the progress in the surgical techniques and the introduction of new anaesthetic drugs. This implies a strict postoperative analgesic strategy in order to avoid both direct and indirect complications associated with pain. In this respect, the use of remifentanil or other techniques like target-controlled injection of opioids should absolutely be considered. In most cases, class I and II analgesics seem to provide optimal pain relief. However, for some patients, the use of an opioid may be required. PMID- 15120791 TI - [Postoperative nausea and vomiting after neurosurgery (infratentorial and supratentorial surgery)]. AB - OBJECTIVE: To perform a synthesis regarding postoperative nausea and vomiting (PONV) after neurosurgery. DATA EXTRACTION: A Medline search was performed to identify publications about frequency, risk factors, prevention and treatment of PONV in adults and children, after neurosurgery. DATA SYNTHESIS: After neurosurgery, the estimated frequency of nausea is around 50% and around 39% for vomiting. After neurosurgery; PONV risk factors are female sex and infratentorial surgery. Children older than two years are at higher risk for PONV. To reduce baseline risk factors, it is recommended to use propofol for induction and maintenance of anaesthesia, to avoid nitrous oxide and to use hydration (20 ml/kg of crystalloids before induction). For PONV prophylaxis, ondansetron and droperidol may be given, using one drug for a moderate risk patient and both drugs for a high-risk patient. Droperidol should not be used in children as a first choice therapy because of an increased risk of extrapyramidal symptoms. Dexamethasone has not been evaluated after neurosurgery. Metoclopramide has no clinically relevant effect for PONV. Especially in neurosurgery, after occurrence of PONV, it is recommended to rule out a possible triggering factor that should need specific treatment. A global management of PONV is proposed, based on the administration of the same drugs given at half the doses used for prophylaxis. PMID- 15120792 TI - [Functional surgery for movement disorders: implications for anaesthesia]. AB - Functional surgery for movement disorders is a recent stereotactic neurosurgical operation, restricted yet to patients with advanced Parkinson's disease or with generalized primary dystonia. One or two electrodes are implanted in the basal ganglia, namely in the globus pallidus pars interna or in the subthalamic nucleus, to realize a deep brain stimulation at high frequency. While this approach needs additional data to demonstrate clinical benefits, first results observed after short and long-term follow up are encouraging. Perioperative problems in patients with Parkinson's disease are possible respiratory disorders, a postoperative miss in medication doses and potential drug interactions with anaesthesia. The objectives of anaesthesia will be to allow stereotactic neurosurgical procedure, to maintain the upper airway patency and to be quickly reversible. PMID- 15120793 TI - [Intra-abdominal pressure measurement using a bladder catheter]. AB - In case of abdomen compartment syndrome, measurement of bladder pressure is an accurate means to assess peritoneal pressure. Authors describe measurement technique and pitfalls to avoid. PMID- 15120794 TI - [Cellulitis of the meso of the rectosigmoid due to primary group A streptococcal peritonitis]. PMID- 15120795 TI - [Unexpected oxygen arrival breakdown]. PMID- 15120796 TI - [Long duration temporary caval filter]. PMID- 15120797 TI - [Unknown profound consequence of previous intubation]. PMID- 15120798 TI - [Tenecteplase and massive pulmonary embolus]. PMID- 15120799 TI - [Spinal anesthesia and ambulatory surgery]. PMID- 15120800 TI - [Account of the 10th workshop of the club d'anesthesie-reanimation en ORL]. PMID- 15120801 TI - [Account of the 17th meeting of the club d'anesthesie-reanimation en ORL Dijon, November 21, 2003]. PMID- 15120802 TI - Smallpox vaccination and myopericarditis: a clinical review. AB - Smallpox is a devastating viral illness that was eradicated after an aggressive, widespread vaccination campaign. Routine U.S. childhood vaccinations ended in 1972, and routine military vaccinations ended in 1990. Recently, the threat of bioterrorist use of smallpox has revived the need for vaccination. Over 450,000 U.S. military personnel received the vaccination between December 2002 and June 2003, with rates of non-cardiac complications at or below historical levels. The rate of cardiac complications, however, has been higher than expected, with two confirmed cases and over 50 probable cases of myopericarditis after vaccination reported to the Department of Defense Smallpox Vaccination Program. The practicing physician should use the history and physical, electrocardiogram, and cardiac biomarkers in the initial evaluation of a post-vaccination patient with chest pain. Echocardiogram, cardiac catheterization, magnetic resonance imaging, nuclear imaging, and cardiac biopsy may be of use in further workup. Treatment is with non-steroidal anti-inflammatory agents, four to six weeks of limited exertion, and conventional heart failure treatment as necessary. Immune suppressant therapy with steroids may be uniquely beneficial in myopericarditis related to smallpox vaccination, compared with other types of myopericarditis. If a widespread vaccination program is undertaken in the future, many more cases of post-vaccinial myopericarditis could be seen. Practicing physicians should be aware that smallpox vaccine-associated myopericarditis is a real entity, and symptoms after vaccination should be appropriately evaluated, treated if necessary, and reported to the Vaccine Adverse Events Reporting System. PMID- 15120803 TI - Angina pectoris prior to myocardial infarction protects against subsequent left ventricular remodeling. AB - OBJECTIVES: To investigate the hypothesis that prior angina pectoris confers protection from remodeling occurring after myocardial infarction (MI), we analyzed echocardiograms from the Healing and Early Afterload Reducing Therapy (HEART) trial. BACKGROUND: Ischemia occurring before MI has been shown to reduce infarct size in experimental models and to improve outcomes in patients. The extent to which ischemia occurring before MI influences subsequent changes in ventricular size and function is unclear. METHODS: We studied 283 patients enrolled in the HEART trial who had echocardiograms at days 1 and 90 after MI. Left ventricular (LV) dilation from days 1 to 90 was used as a measure of LV remodeling. We explored the relationship between symptomatic angina occurring before infarction and subsequent LV remodeling. RESULTS: In patients who reported angina (n = 111) during the three months preceding MI, LV volume change was -0.73 +/- 2.6 ml over the 90-day post-MI period, compared with 6.8 +/- 2.6 ml for patients (n = 172) without angina (p = 0.017). In contrast, there were no differences in changes in ejection fraction based on prior angina. Maximal creatine kinase was significantly lower in patients with prior angina (2,119 +/- 1,729 vs. 2,701 +/- 2,088, p = 0.016). In a multivariate model, prior angina remained protective for ventricular remodeling after adjusting for age, gender, baseline ejection fraction, Killip class, baseline end-diastolic volume, and drug treatment group (p = 0.042). However, the protective effect of pre-infarction angina appeared to be attenuated in diabetic patients. CONCLUSIONS: Ischemic symptoms occurring before MI may protect against LV remodeling. These protective effects may be secondary to recruitment of collaterals or ischemic preconditioning of the myocardium, and they appear to be attenuated in diabetic patients. PMID- 15120804 TI - Ischemic preconditioning in coronary heart disease: a therapeutic golden fleece? PMID- 15120805 TI - Cardiology management improves secondary prevention measures among patients with coronary artery disease. AB - OBJECTIVES: The goal of this study was to determine if cardiology subspecialty involvement improves the attainment of recommended low-density lipoprotein (LDL) cholesterol and blood pressure (BP) targets in coronary artery disease (CAD) patients. BACKGROUND: The impact of physician specialty on secondary prevention measures for CAD in ambulatory care is unknown. METHODS: This was a retrospective cohort study of 13,995 patients with CAD seen at eight ambulatory care Veteran Affairs facilities from 1998 to 2000. Patients with cardiology involvement were defined as those seen in cardiology clinic in addition to primary care. The main outcomes of interest were LDL cholesterol < or =100 mg/dl and BP < or =130/85 mm Hg. Multivariable hierarchical regression analyses were used to determine the independent association of cardiology involvement with improved LDL cholesterol and BP control. RESULTS: Overall, 3,771 (27.0%) patients had cardiology involvement. A higher proportion of patients with cardiology involvement achieved LDL cholesterol (55.6% vs. 45.6%; p < 0.01) and BP (45.3% vs. 35.9%; p < 0.01) goals. In multivariable hierarchical regression analysis, cardiology involvement was independently associated with better LDL cholesterol (odds ratio [OR], 1.59; 95% confidence interval [CI], 1.40 to 1.82) and BP (OR, 1.52; 95% CI, 1.32 to 1.77) control. The benefit of cardiology involvement was consistent across a range of LDL and BP targets, in analysis of LDL and BP as continuous outcomes, and among subgroups of high-risk patients, including diabetic patients, the elderly, and those with prior revascularization. CONCLUSIONS: Cardiology involvement is associated with better LDL cholesterol and BP control among CAD patients. However, significant room for improvement in secondary prevention measures remains, irrespective of physician specialty. PMID- 15120806 TI - Limitation of exercise tolerance in chronic heart failure: distinct effects of left bundle-branch block and coronary artery disease. AB - OBJECTIVES: The aim of this study was to identify resting measurements of left ventricular (LV) function that predict exercise capacity in dilated cardiomyopathy (DCM); in particular, the effects of left bundle branch block (LBBB), coronary artery disease (CAD), and total isovolumic time (t-IVT). BACKGROUND: The t-IVT is a major determinant of cardiac output during dobutamine stress in DCM, and is itself determined by the presence or absence of LBBB and CAD. METHODS: A total of 111 patients with DCM, 51 with CAD (29 LBBB), and 60 without CAD (30 LBBB) were studied with echocardiography and cardiopulmonary exercise testing. The t-IVT (in s/min) was measured by Doppler echocardiography, and maximal oxygen consumption (peak Vo(2)) and percentage of the normal predicted peak Vo(2) (%predicted peak Vo(2)) were obtained from exercise testing. RESULTS: Left bundle branch block reduced peak Vo(2) (by 10.5 ml.kg(-1)min(-1)) and %predicted peak Vo(2) (by 33%, both p < 0.001) compared with patients without LBBB. Coronary artery disease reduced peak Vo(2) (by 5.5 ml.kg(-1)min(-1), p < 0.001) and %predicted peak Vo(2) (by 14%, p < 0.01) compared with those without CAD (p < 0.01). The t-IVT, CAD, LBBB, and QRS duration were univariate predictors of exercise tolerance, but only t-IVT and CAD were independent predictors. The t IVT at rest correlated with peak Vo(2) (r = -0.68) and %predicted peak Vo(2) (r = -0.74, both p < 0.001). The combination of t-IVT and CAD explained 57% (r = 0.75, p < 0.001) of the total variance in exercise capacity. CONCLUSIONS: Resting t-IVT and less prominently, CAD, are major determinants of exercise tolerance in DCM. Left bundle branch block significantly determines resting t-IVT and thus peak Vo(2). Prediction of maximum exercise capacity in DCM is therefore possible from time-domain analysis of LV function at rest. PMID- 15120807 TI - Understanding the impact of abnormal cardiac activation on cardiac function. PMID- 15120808 TI - Predischarge initiation of carvedilol in patients hospitalized for decompensated heart failure: results of the Initiation Management Predischarge: Process for Assessment of Carvedilol Therapy in Heart Failure (IMPACT-HF) trial. AB - OBJECTIVES: The Initiation Management Predischarge: Process for Assessment of Carvedilol Therapy in Heart Failure (IMPACT-HF) trial was an investigator initiated study to evaluate if predischarge carvedilol initiation in stabilized patients hospitalized for heart failure (HF) increased the number of patients treated with beta-blockade at 60 days after randomization without increasing side effects or length of hospital stay. BACKGROUND: Beta-blockers are underused in HF. Predischarge initiation may improve the use of evidence-based beta-blockade. METHODS: The IMPACT-HF was a prospective, randomized open-label trial conducted in 363 patients hospitalized for HF. Patients were randomized to carvedilol initiation pre-hospital discharge or to postdischarge initiation (>2 weeks) of beta-blockade at the physicians' discretion. The primary end point of the study was the number of patients treated with beta-blockade at 60 days after randomization. Secondary end points included the number of patients discontinuing beta-blockade, median dose achieved, and a composite of death, rehospitalization, unscheduled visit for HF, or > or =50% increase in oral diuretic, new oral diuretic, or any intravenous therapy with diuretics, inotropes, or other vasoactive agents. RESULTS: At 60 days 165 patients (91.2%) randomized to predischarge carvedilol initiation were treated with a beta-blocker, compared with 130 patients (73.4%) randomized to initiation postdischarge (p < 0.0001). Predischarge initiation was not associated with an increased risk of serious adverse events. The median length of stay was five days in both groups. CONCLUSIONS: Predischarge initiation of carvedilol in stabilized patients hospitalized for HF improved the use of beta-blockade at 60 days without increasing side effects or length of stay. Predischarge initiation may be one approach to improve beta-blocker use in this population. PMID- 15120810 TI - Depression: are we ignoring an important comorbidity in heart failure? PMID- 15120809 TI - The influence of age, gender, and race on the prevalence of depression in heart failure patients. AB - OBJECTIVES: The goal of this study was to determine the prevalence of depression in an out-patient heart failure (HF) population; its relationship to quality of life (QOL); and the impact of gender, race, and age. BACKGROUND: Most studies of depression in HF have evaluated hospitalized patients (a small percentage of the population) and have ignored the influence of various patient characteristics. Although reported depression rates among hospitalized patients range from 13% to 77.5%, out-patient studies have been small, have reported rates of 13% to 42%, and have not adequately accounted for the impact of age, race, or gender. METHODS: A total of 155 patients with stable New York Heart Association functional class II, III, and IV HF and an ejection fraction <40% were given questionnaires to assess QOL and depression. These included the Medical Outcomes Study Short Form, the Minnesota Living with Heart Failure questionnaire, and the Beck Depression Inventory (BDI). Depression was defined as a score on the BDI of > or =10. RESULTS: A total of 48% of the patients scored as depressed. Depressed patients tended to be younger than non-depressed patients. Women were more likely (64%) to be depressed than men (44%). Among men, blacks (34%) tended to have less depression than whites (54%). Depressed patients scored significantly worse than non-depressed patients on all components of both the questionnaires measuring QOL. However, they did not differ in ejection fraction or treatment, except that depressed patients were significantly less likely to be receiving beta-blockers. CONCLUSIONS: Depression is common in patients with HF, with age, gender, and race influencing its prevalence in ways similar to those observed in the general population. These data suggest that pharmacologic or non-pharmacologic treatment of depression might improve the QOL of HF patients. PMID- 15120811 TI - Outcomes in cardiac transplant recipients using allografts from older donors versus mortality on the transplant waiting list; Implications for donor selection criteria. AB - OBJECTIVES: This study investigates the outcomes of cardiac transplantation using older donors. BACKGROUND: Despite high mortality rates on waiting lists, transplanting hearts from older donors remains a relative contraindication. METHODS: We retrospectively reviewed data on 479 adult heart transplant recipients, 352 status I patients, and 534 status II patients enrolled on a waiting list between 1992 and 1999. The Cox proportional hazards model was used for statistical analysis. RESULTS: Of all donors, 20% were 40 to 50 years old and 8% were > or =50 years old. The risk of six-month mortality on the waiting list for patients who were not transplanted (status I: relative risk [RR] 8.5; status II: RR 3.7) significantly outweighed the risk of transplanting patients with a heart from donors >40 years old (status I: RR 1.6; status II: RR 2.1). Recipients of cardiac allografts from donors <40 years old had a one-month mortality rate of 5%, in contrast to 13% and 22% in those receiving allografts from donors 40 to 50 years old and > or =50 years old, respectively. Donor age did not influence long term survival or frequency of rejections; however, it did correlate with the early presence of transplant-related coronary artery disease (TCAD). By the first annual angiogram, only 17% of recipients with donors <20 years old developed TCAD, in contrast to 26% to 30% and 34% of recipients who received allografts from donors age 20 to 40 years and >40 years, respectively. CONCLUSIONS: Despite a strong association between older donor age and increased post-operative mortality and TCAD, it is more beneficial in terms of patient survival to receive an allograft from a donor >40 years old than to remain on the waiting list. PMID- 15120812 TI - Adverse outcomes from the use of older donor hearts in cardiac transplant recipients: the pros and cons of expanded donor criteria. PMID- 15120814 TI - Left ventricular assist device malfunction: a systematic approach to diagnosis. AB - OBJECTIVES: A protocol was designed to diagnose the common malfunctions of a left ventricular assist device (LVAD). BACKGROUND: Mechanical circulatory support, primarily with an LVAD, is increasingly used for treatment of advanced heart failure (HF). Left ventricular assist device dysfunction is a recognized complication; but heretofore, a systematic method to accurately diagnose LVAD dysfunction has not been thoroughly described. METHODS: We developed a catheter based protocol designed to characterize a normally functioning LVAD and diagnose multiple types of dysfunction. A total of 15 studies of 10 patients supported with an LVAD were reviewed. All patients had been evaluated due to concerns regarding LVAD dysfunction. RESULTS: Of 15 examinations performed, 11 documented severe LVAD inflow valve regurgitation. One of these cases proved to have coexistent severe mitral valve regurgitation. One case was diagnosed with distortion of the LVAD outflow graft. One case of suspected embolization from the pumping chamber excluded the outflow graft as the source of emboli. One study had aortic insufficiency. CONCLUSIONS: As LVAD use for treatment of end-stage HF becomes widespread and durations of support are extended, dysfunction will be increasingly prevalent. This catheter-based protocol provided a practical method to diagnose multiple causes of LVAD dysfunction. PMID- 15120813 TI - Angiotensin II receptors from peritransplantation through first-year post transplantation and the risk of transplant coronary artery disease. AB - OBJECTIVES: We evaluated whether the angiotensin II (Ang II) receptors from perioperation through one-year post-transplantation predict the transplant coronary artery disease (TCAD) progression. BACKGROUND: The role of Ang II receptors (type 1: AT(1)R; type 2: AT(2)R) in TCAD is uncertain. METHODS: We investigated 28 heart donors and the corresponding recipients. The levels of AT(1)R and AT(2)R messenger ribonucleic acid (mRNA) were examined in lymphocytes from the donor spleen and in the donor heart at one-week and one-year posttransplantation to determine their association with the progression of TCAD, measured as changes in maximal intimal thickness (CMIT) and plaque volume (CPV) by intravascular ultrasound (IVUS) examinations. RESULTS: The AT(1)R mRNA in lymphocytes from the donor spleen (CMIT: r = 0.73, p < 0.0001; CPV: r = 0.69, p < 0.0001) and in the donor hearts at one-week (CMIT: r = 0.52, p = 0.005; CPV: r = 0.56, p = 0.002) and at one-year (CMIT: r = 0.63, p < 0.0001; CPV: r = 0.43, p = 0.004) post-transplantation along with AT(2)R mRNA in the donor hearts at one year post-transplantation (CMIT: r = 0.3, p < 0.0001; CPV: r = 0.53, p = 0.009) were univariate predictors, whereas AT(1)R mRNA in lymphocytes and in the donor hearts at one-year post-transplantation proved to be multivariate predictors of the progression of TCAD. CONCLUSIONS: These data suggest a role for Ang II receptors in the pathogenesis of TCAD and support a novel concept that TCAD may have its origin in the donor per se and may be modulated by the recipient's inherent biological factors. PMID- 15120815 TI - B-type natriuretic peptide and peak exercise oxygen consumption provide independent information for risk stratification in patients with stable congestive heart failure. AB - OBJECTIVES: The aim of this study was to compare the prognostic value of peak oxygen consumption (VO(2)) and B-type natriuretic peptide (BNP) in patients with stable congestive heart failure (CHF). BACKGROUND: Previous studies have demonstrated that both peak VO(2) and BNP are useful for risk stratification in patients with CHF. No study has compared the respective prognostic value of these two parameters in a large series of patients receiving a combination of angiotensin-converting enzyme inhibitors and of beta-blockers. METHODS: Patients with stable CHF underwent radionuclide angiography, echocardiography, 24-h Holter monitoring, and a cardiopulmonary exercise test. Blood samples were drawn for standard measurements and for hormonal determinations. RESULTS: After a median follow-up period of 787 days, there were 75 cardiac-related deaths and three urgent transplantations. Independent predictors of cardiac survival were percent of maximal predicted VO(2) (%VO(2), relative risk [RR] = 2.84 [95% confidence interval, CI = 1.73 to 4.65], p < 0.00001), BNP (RR = 3.17 [95% CI 1.68 to 5.96], p = 0.0004), left atrial diameter (LAD) (RR = 2.04 [95% CI 1.25 to 3.34], p = 0.004), age (RR = 1.93 [95% CI 1.22 to 3.05], p = 0.005), and aldosterone (RR = 1.84 [95% CI 1.12 to 3.00], p = 0.015). In patients with infra-median levels of BNP (<109 pg/ml), age was the only independent predictor of cardiac survival. However, in patients with supra-median levels of BNP, independent predictors of cardiac survival were %VO(2) (RR = 3.76 [95% CI 2.19 to 6.45], p < 0.00001) and LAD (RR = 1.90 [95% CI 1.10 to 3.28], p = 0.02). CONCLUSIONS: B-type natriuretic peptide, in combination with %VO(2), improves risk stratification of patients with stable CHF. PMID- 15120816 TI - Obesity and suppressed B-type natriuretic peptide levels in heart failure. AB - OBJECTIVES: This investigation evaluated the relationship between obesity and B type natriuretic peptide (BNP) in heart failure. BACKGROUND: Obesity is a major risk factor for the development of heart failure, but the precise mechanisms remain uncertain. Physiologically, natriuretic peptides and lipolysis are closely linked. METHODS: A total of 318 patients with heart failure were evaluated between June 2001 and June 2002. Levels of BNP were compared in obese (body mass index [BMI] > or =30 kg/m(2)) and nonobese (BMI <30 kg/m(2)) patients with respect to New York Heart Association functional class and lean body weight adjusted peak aerobic oxygen consumption. In a subset of 36 patients, plasma levels of tumor necrosis factor-alpha, interleukin-6, and soluble intercellular adhesion molecule-1 were measured. RESULTS: The population's BMI was 29.4 +/- 6.6 kg/m(2); 24% were lean (BMI <25 kg/m(2)), 31% overweight (BMI > or =25 to 29.9 kg/m(2)), and 45% obese (BMI > or =30 kg/m(2)). Obese patients were younger, more often African American, and more likely to have a history of antecedent hypertension, but less likely to have coronary artery disease and with only a trend toward diabetes mellitus. Levels of BNP were significantly lower in obese than in nonobese subjects (205 +/- 22 and 335 +/- 39 pg/ml, respectively; p = 0.0007), despite a similar severity of heart failure and cytokine levels. Multivariate regression analysis identified BMI as an independent negative correlate of BNP level. There were no differences in emergency department visits, heart failure hospitalization, or death between the obese and nonobese patients at 12-month follow-up. CONCLUSIONS: Our investigation indicates that a state of reduced natriuretic peptide level exists in the obese individual with heart failure. PMID- 15120817 TI - Intracranial hemorrhage and hyperperfusion syndrome following carotid artery stenting: risk factors, prevention, and treatment. AB - OBJECTIVES: The study defined the incidence of cerebral hyperperfusion syndrome and intracranial hemorrhage (ICH) and the risk factors for their development following carotid artery stenting (CAS). BACKGROUND: Hyperperfusion syndrome and ICH can complicate carotid revascularization, be it endarterectomy or CAS. Although extensive effort has been devoted to reducing the incidence of ischemic stroke complicating CAS, little is known about the incidence, etiology, and prevention strategies for hyperperfusion and ICH following CAS. METHODS: We retrospectively reviewed the prospective database of 450 consecutive patients who were treated with CAS in our department to identify patients who developed hyperperfusion syndrome and/or ICH. RESULTS: The mean age of the patients was 72.7 +/- 10.9 years, and the mean diameter narrowing was 84 +/- 12.8%. Five (1.1% [95% confidence interval 0.4% to 2.6%]) patients developed hyperperfusion. Three (0.67%) of the five developed ICH. Two of these patients died (0.44%). Symptoms developed within a median of 10 h (range, 6 h to 4 days) following stenting. All five patients had correction of a severe internal carotid stenosis (mean 95.6 +/- 3.7%) with a concurrent contralateral stenosis >80% or contralateral occlusion and peri-procedural hypertension. These same risk factors are involved in cerebral hyperperfusion following carotid endarterectomy. The use of platelet glycoprotein IIb/IIIa receptor blockers did not appear to increase the risk ICH. CONCLUSIONS: The hyperperfusion syndrome occurs infrequently following CAS, and ICH occurs in 0.67% of patients. Patients with severe bilateral carotid stenoses may be predisposed to ICH, particularly if there is concurrent arterial hypertension. Patients with these factors may require more intensive hemodynamic monitoring after CAS, including prolongation of hospitalization in some cases. PMID- 15120818 TI - A cardiologist in the carotids. AB - Carotid endarterectomy for stroke prevention has been the standard of care for 50 years in patients with extra-cranial carotid bifurcation disease. Over the past decade, carotid stenting has emerged as a viable alternative to surgery. Combined with filter embolic protection devices, both a randomized control trial (Stenting and Angioplasty with Protection in Patients at High Risk for Endarterectomy registry [SAPPHIRE]) as well as registry data (ACCULINK for Revascularization of Carotids in High Risk Patients registry [ARCHeR] and Registry Study to evaluate the Neuroshield Bare-Wire Cerebral Protection System and X-Act Stent in patients at high risk for Carotid Endarterectomy [SECuRITY]) have compared favorably to endarterectomy in patients at high risk for operative revascularization. Conditions associated with high operative risk included patients with significant cardiac, pulmonary, and renal disease; previous neck operation; previous radiation; and anatomically difficult surgical access. On the basis of these results, a carotid stent system approved by the Food and Drug Administration (FDA) is anticipated in 2004. Although this will be a welcome addition to endarterectomy in the armamentarium of therapeutic options for patients with carotid disease, several challenges lie ahead. Coverage and reimbursement for the carotid stenting has been severely restricted to include only those procedures performed as part of an FDA investigational device exemption trial protocol, and a national noncoverage decision will have to be reckoned with before broader coverage can be put into place (assuming FDA approval). In addition, the level of national expertise in carotid endovascular intervention is limited, and training will need to be tailored to the three specialties likely to perform the procedure: cardiology, radiology, and vascular surgery. Each of these specialties will have specific, and different, requirements for their training, further complicating the task of education. PMID- 15120819 TI - The profile of cardiac patients with renal artery stenosis. AB - OBJECTIVES: We examined the prevalence and severity of renal artery stenosis (RAS) in patients undergoing cardiac catheterization who were deemed at risk for RAS based on clinical or laboratory criteria for study entry, but who had not previously been suspected of having RAS. BACKGROUND: The diagnosis of atherosclerotic RAS remains problematic because its clinical manifestations are nonspecific. METHODS: Consecutive patients undergoing non-emergent cardiac catheterization at a single institution during a 12-month period were evaluated using standardized clinical, laboratory, and angiographic criteria. Patients exhibiting at least one of four predefined selection criteria (severe hypertension, unexplained renal dysfunction, acute pulmonary edema with hypertension, or severe atherosclerosis) were prospectively registered and underwent coincident diagnostic renal angiography. RESULTS: Renal angiography was performed in 851 patients and was diagnostic in 837. Angiographically evident renal atherosclerosis was present in 39% of the population, with RAS > or =50% in 120 (14.3%) and severe stenosis (> or =70%) in 61 (7.3%). Severe stenosis was present in 48 (7%) patients with severe atherosclerosis, 38 (16%) with renal dysfunction, 25 (9%) with hypertension, and 2 (22%) with acute pulmonary edema with hypertension. The prevalence was higher in those exhibiting multiple selection criteria. In a multivariate model, severe RAS was associated with age, female gender, reduced creatinine clearance, increased systolic blood pressure, and peripheral or carotid artery disease. CONCLUSIONS: In a population at risk of, but not previously suspected of having RAS, severe RAS is associated with simple and readily determined clinical and laboratory patient characteristics. These data facilitate focused application of diagnostic renal angiography. PMID- 15120820 TI - Renal artery stenosis: "fortuitous diagnosis," problematic therapy. PMID- 15120821 TI - Predictors of stroke in patients paced for sick sinus syndrome. AB - OBJECTIVES: This study was an analysis of factors associated with stroke in a population of patients paced for sinus node dysfunction in a large prospective clinical trial (Mode Selection Trial [MOST]). BACKGROUND: The effects of dual chamber versus single-chamber ventricular pacing on subsequent stroke in patients with sinus node dysfunction are not known. METHODS: A total of 2,010 patients with sinus node dysfunction were randomized to ventricular or dual-chamber pacing and followed for a median of 33.1 months. RESULTS: The median participant age was 74 years. During 5,664 patient-years of follow-up, 90 strokes (11 hemorrhagic) occurred. By life-table analysis, the rate of stroke was 2.2% (95% confidence interval [CI] 1.6 to 2.9) at one year and 5.8% (95% CI 4.5 to 7.1) at four years. The incidence of stroke was not significantly different in dual-chamber (4%) as compared with ventricular-paced patients (4.9%) (hazard ratio [HR] 0.82, 95% CI 0.54 to 1.25, p = 0.36). Multivariable analysis demonstrated that significant predictors of stroke included prior stroke or transient ischemic attack, Caucasian race, hypertension, prior systemic embolism, and New York Heart Association functional class III or IV (p < 0.05); pacing mode remained non significant after adjustment for these factors (p = 0.37). Clinically reported atrial fibrillation after implantation was a risk factor for stroke in this cohort after adjustment for other predictors of stroke (p = 0.042, HR 1.68 [95% CI 1.02 to 2.76]). CONCLUSIONS: Clinical characteristics, but not mode of pacing, were associated with subsequent stroke in patients paced for sinus node dysfunction. PMID- 15120822 TI - Stroke and pacing mode: is pacing mode important? PMID- 15120824 TI - Predictors of new malignant ventricular arrhythmias after coronary surgery: a case-control study. AB - OBJECTIVES: We sought to investigate the relationship between perioperative factors and the occurrence of ventricular tachycardia (VT) and ventricular fibrillation (VF), as well as the impact of VT/VF on early and late mortality. BACKGROUND: Both VT and VF are rare but serious complications after coronary artery bypass graft surgery (CABG), and their etiology and implications remain uncertain. METHODS: Data on 4,411 consecutive patients undergoing CABG (1,154 [25.8%] had off-pump surgery) between April 1996 and September 2001 were extracted from a prospective database and analyzed. Odds ratios (ORs) describing associations between possible risk factors and VT/VF were estimated separately. Factors observed to be significantly associated with VT/VF were further investigated using multivariate logistic regression. RESULTS: Sixty-nine patients suffered VT/VF (1.6%). There were 61 (1.4%) in-hospital/30-day deaths, 15 among patients who had postoperative VT/VF (21.7%). Patient factors independently associated with an increase in the odds of VT/VF included age <65 years, female gender, body mass index <25 kg/m(2), unstable angina, moderate or poor ejection fraction, and the need for inotropes and an intra-aortic balloon pump (OR 1.72 to 4.47, p < 0.05). After adjustment, off-pump surgery was associated with a substantial but nonsignificant protective effect against VT/VF (OR 0.53, 95% confidence interval [CI] 0.25 to 1.13; p = 0.10). Actuarial survival at two years was 98.2% among patients who had VT/VF and who survived to discharge/30 days, compared with 97.0% for the control group (adjusted hazard ratio 0.96 (95% CI 0.40 to 2.31, p = 0.92). CONCLUSIONS: The incidence of VT/VF is low in patients undergoing coronary surgery but is associated with high in-hospital mortality. The late survival of the discharged VT/VF patients compares favorably with that of controls. PMID- 15120823 TI - Postmortem molecular screening in unexplained sudden death. AB - OBJECTIVES: We examined the prevalence of defects in arrhythmia-related candidate genes among patients with unexplained sudden cardiac death (SCD). BACKGROUND: Patients with unexplained sudden death may constitute up to 5% of overall SCD cases. For such patients, systematic postmortem genetic analysis of archived tissue, using a candidate gene approach, may identify etiologies of SCD. METHODS: We performed analysis of KCNQ1 (KVLQT1), KCNH2 (HERG), SCN5A, KCNE1, and KCNE2 defects in a subgroup of 12 adult subjects with unexplained sudden death, derived from a 13-year, 270-patient autopsy series of SCD. Archived, paraffin-embedded myocardial tissue blocks obtained at the original postmortem examination were the source of deoxyribonucleic acid for genetic analysis. RESULTS: Two patients were found to have the same HERG defect, a missense mutation in exon 7 (nucleotide change G1681A, coding effect A561T). The mutation was heterozygous in Patient 1, but Patient 2 appeared to be homozygous for the defect. Patch-clamp recordings showed that the A561T mutant channel expressed in human embryonic kidney cells failed to generate HERG current. Western blot analysis implicated a trafficking defect in the protein, resulting in loss of post-translational processing from the immature to the mature form of HERG. No mutations were detected among the remaining four candidate genes. CONCLUSIONS: In this autopsy series, only 2 of 12 patients with unexplained sudden death were observed to have a defect in HERG among five candidate genes tested. It is likely that elucidation of SCD mechanisms in such patients will await the discovery of multiple, novel arrhythmia-causing gene defects. PMID- 15120825 TI - Functional characterization of the crista terminalis in patients with atrial flutter: implications for radiofrequency ablation. AB - OBJECTIVES: The aim of the study was to investigate the conduction properties and anisotropy of the crista terminalis (CT) in patients with atrial flutter (AFL) using non-contact mapping. BACKGROUND: The CT is a posterior barrier during typical AFL. However, the CT has transverse conduction capabilities in patients with upper loop re-entry (ULR). METHODS: Twenty-two patients (16 males, 63 +/- 15 years) with typical AFL and ULR were included. Non-contact mapping of the right atrium during AFL and pacing from coronary sinus (CS) and low anterolateral right atrium (LARA) was performed to evaluate transverse conduction across the CT. During ULR, the longitudinal (CV(L)) and transverse (CV(T)) conduction velocity along and across the CT were measured. The width of the CT conduction gap was evaluated to guide radiofrequency ablation (RFA). RESULTS: No transverse CT gap conduction was found during typical AFL. Transverse CT gap conduction was found in three patients during CS pacing and in three patients during LARA pacing. During ULR, CV(L) was greater than CV(T) (1.28 +/- 0.43 vs. 0.73 +/- 0.30 m/s, p < 0.001). The CV(L)/CV(T) ratio was 1.95 +/- 0.77, which was inversely related to the CT gap width (15.7 +/- 6.8 mm) (p < 0.001). The RFA of the CT gap was successful in 18 patients. Four patients had recurrence of arrhythmias during the follow-up of 11 +/- 3 months. CONCLUSIONS: Most of the CT conduction gaps were functional and only appeared during ULR. The width of the CT gap was inversely related to the anisotropic ratio of the CT. The RFA of the CT gap was effective in eliminating ULR. PMID- 15120826 TI - Doppler-derived mitral deceleration time as a strong prognostic marker of left ventricular remodeling and survival after acute myocardial infarction: results of the GISSI-3 echo substudy. AB - OBJECTIVES: The goal of this study was to assess the impact of left ventricular (LV) diastolic filling on remodeling and survival after acute myocardial infarction (AMI). BACKGROUND: Little is known regarding the link between LV filling, its changes over time, and six-month remodeling and late survival in uncomplicated AMI. METHODS: Doppler mitral profile, end-diastolic volume index (EDVi) and end-systolic volume index (ESVi), ejection fraction (EF), and wall motion abnormalities (%WMA) were evaluated in 571 patients from the GISSI-3 Echo substudy at baseline, pre-discharge, and six months after AMI. Patients with baseline early mitral deceleration time (DT) 130 ms were assigned to the restrictive group (n = 147), and those with DT >130 ms to the nonrestrictive group (n = 424). RESULTS: Restrictive group patients had greater baseline ESVi and %WMA and lower EF than nonrestrictive group, and six-month greater LV dilation (EDVi, ESVi: p < 0.001 for EDVi and ESVi), smaller decrease in %WMA decrease (p < 0.01), and larger EF impairment (p < 0.008). Among the restrictive group, patients (n = 56) with pre-discharge persistent restrictive filling (n = 56) showed six-month greater LV enlargement (p < 0.001) and EF impairment (p < 0.009) than those (n = 91) with reversible restrictive filling. Baseline %WMA and EDVi, together with pre-discharge persistent restrictive filling, predicted severe (>20%) LV dilation. Four-year survival was 93% in nonrestrictive patients versus 88% in the restrictive group (p < 0.06), and 93% in pre-discharge reversible restrictive versus 79% in persistent restrictive (p < 0.0003). The single best predictor of mortality, by Cox analysis, was pre-discharge persistent restrictive filling (chi-square 14.88). CONCLUSIONS: Left ventricular dilation may occur even after uncomplicated AMI and may be paralleled by an improvement in LV filling. However, a baseline restrictive filling that persists at pre discharge identifies more compromised patients at higher risk for six-month remodeling and four-year mortality. PMID- 15120827 TI - Noninvasive assessment of ejection intraventricular pressure gradients. AB - OBJECTIVES: The study was designed to validate in vivo a new method to measure ejection intraventricular pressure gradients (IVPGs) by processing color M-mode Doppler data and to assess the effects of inotropic interventions on IVPGs in the clinical setting. BACKGROUND: In the absence of obstruction, ejection IVPGs cannot be estimated by Doppler using the simplified Bernoulli equation. METHODS: High-fidelity micromanometers were placed in the left ventricle of eight minipigs, and synchronic Doppler images and pressure signals were obtained during different hemodynamic conditions. Twenty healthy volunteers and 20 dilated cardiomyopathy patients were studied at baseline and during esmolol, dobutamine, and atropine infusion (only dobutamine in patients). RESULTS: Excellent agreement was observed between micromanometer and Doppler methods for measuring instantaneous pressure differences among the apex, the mid-cavity, and the outflow tract (R(intraclass) = 0.98, 0.81, 0.76, and 0.98 for the peak, time-to peak, peak reverse, and time-to-peak reverse values, respectively; n = 810 beats). Error of the noninvasive method was -0.05 +/- 0.25 mm Hg for the peak pressure difference. Parametrical images demonstrated that IVPGs originate mainly in the mid-ventricle and then propagate to the outflow tract. Both the magnitude and the temporal course of IVPGs were different among volunteers and patients. Inotropic interventions induced significant changes in the apex-outflow tract pressure differences in both populations, whereas atropine had no effect on IVPGs. CONCLUSIONS: For the first time, ejection IVPGs can be accurately visualized and measured by Doppler-echocardiography. Important aspects of the dynamic interaction among myocardial performance, load mechanics, and ejection dynamics can be assessed in the clinical setting using this method. PMID- 15120828 TI - Prognostic value of coronary artery calcium screening in subjects with and without diabetes. AB - OBJECTIVES: The study was done to determine the interaction of coronary artery calcium and diabetes mellitus for prediction of all-cause death. BACKGROUND: Diabetes is a strong risk factor for coronary artery disease (CAD) and is associated with an elevated overall mortality. Electron beam tomography (EBT) provides information on the presence of subclinical atherosclerosis and may be useful for risk stratification. METHODS: We followed 10,377 asymptomatic individuals (903 diabetic patients) referred for EBT imaging. Primary end point was all-cause mortality, and the average follow-up was 5.0 +/- 3.5 years. Cox proportional hazard models, with and without adjustment for other risk factors, were developed to predict all-cause mortality. RESULTS: Patients with diabetes had a higher prevalence of hypertension and smoking (p < 0.001) and were older. The average coronary calcium score (CCS) for subjects with and for those without diabetes was 281 +/- 567 and 119 +/- 341, respectively (p < 0.0001). Overall, the death rate was 3.5% and 2.0% for subjects with and without diabetes (p < 0.0001). In a risk-factor-adjusted model, there was a significant interaction of CCS with diabetes (p < 0.00001), indicating that, for every increase in CCS, there was a greater increase in mortality for diabetic than for nondiabetic subjects. However, patients suffering from diabetes with no coronary artery calcium demonstrated a survival similar to that of individuals without diabetes and no detectable calcium (98.8% and 99.4%, respectively, p = 0.5). CONCLUSIONS: Mortality from all causes is increased in asymptomatic patients with diabetes in proportion to the screening CCS. Nonetheless, subjects without coronary artery calcium have a low short-term risk of death even in the presence of diabetes mellitus. PMID- 15120829 TI - Neoangiogenesis, T-lymphocyte infiltration, and heat shock protein-60 are biological hallmarks of an immunomediated inflammatory process in end-stage calcified aortic valve stenosis. AB - OBJECTIVES: We investigated the main biomolecular features in the evolution of aortic stenosis, focusing on advanced lesions. BACKGROUND: "Degenerative" aortic valve stenosis shares risk factors and inflammatory similarities with atherosclerosis. METHODS: We compared nonrheumatic stenotic aortic valves from 26 patients undergoing surgical valve replacement (group A) and 14 surgical control patients (group B). We performed semiquantitative histological and immunohistochemical analyses on valve leaflets to measure inflammation, sclerosis, calcium, neoangiogenesis, and intercellular adhesion molecule-1 (ICAM 1) and vascular cell adhesion molecule-1 (VCAM-1) expression. We assessed heat shock protein 60 (hsp60) gene expression as an index of cellular stress and C reactive protein, erythrocyte sedimentation rate, and fibrinogen as systemic inflammatory markers. RESULTS: In group A valves, we found a prevalence of calcium nodules surrounded by activated inflammatory infiltrates, neovessels, and abundant ICAM-1, VCAM-1, and hsp60 gene expression. Specimens from group B were negative for all of these markers, except 2 of 14 positivity for hsp60. The presence of active inflammatory infiltrates correlated with an abundance of thin neovessels (p < 0.01) and hsp60 gene expression (p = 0.01), whereas neoangiogenesis correlated with inflammation (p = 0.04), calcium (p = 0.01), and hsp60 gene expression (p = 0.04). CONCLUSIONS: "Degenerative" aortic valve stenosis appears to be a chronic inflammatory process associated with atherosclerotic risk factors. The coexistence of neoangiogenesis, T-lymphocyte infiltration, adhesion molecules, and hsp60 gene expression indicates an active immunomediated process in the final phases of the disease. PMID- 15120830 TI - Reversible atrioventricular block associated with closure of atrial septal defects using the Amplatzer device. AB - OBJECTIVES: We sought to determine the incidence, nature, and predisposing factors of atrioventricular block (AVB) associated with closure of atrial septal defects (ASDs) using the Amplatzer septal occluder (ASO). BACKGROUND: In our institution, 162 patients underwent ASD closure using ASO between December 1997 and December 2001. This includes small children with large defects. METHODS: Electrocardiographic tracings during ASO implantation and at follow-up visits were reviewed. Anatomic characteristics and device size were assessed as potential risk factors for AVB. RESULTS: Ten patients (6.2%) presented with new onset (n = 9) or aggravation of preexisting (n = 1) AVB. Atrioventricular block occurred during the procedure (n = 3) or was first noted one day to one week later (n = 7). Patients had first-degree (n = 4), second-degree Wenckebach (n = 4), or third-degree (n = 2) AVB, with no symptoms or hemodynamic compromise. First-degree AVB persisted in two patients at 12 and 33 months of follow-up, whereas most recovered normal AV conduction within one (n = 7) or six months (n = 1). A larger shunt (Qp/Qs ratio 2.8 +/- 0.9 vs. 2.1 +/- 0.8, p < 0.01) and device size (24 +/- 5 vs. 19 +/- 6 mm, p < 0.01) were the only determinant factors for AVB. A device size > or =19 mm was used in 90% (9 of 10) of patients who developed AVB, as compared with 49% of those without AVB (p < 0.02). CONCLUSIONS: Closure of ASDs using the large ASO can be associated with the development of AV block and mandate a closer follow-up. In our series, however, all AVBs resolved or improved spontaneously, with no recurrence at mid-term follow-up. PMID- 15120831 TI - Preservation of ischemic myocardial function and integrity with targeted cytoskeleton-specific immunoliposomes. AB - OBJECTIVES: We sought to demonstrate preservation of myocardial function and integrity after targeted cytoskeleton-specific immunoliposome (CSIL) treatment of globally ischemic Langendorff instrumented hearts and a time response to treatment. BACKGROUND: Cell membrane lesion sealing of hypoxic cardiocytes in culture with CSIL has been reported. METHODS: Langendorff-perfused isolated rat hearts were subjected to global ischemia (25 min). Either CSIL or placebo administration (1-min ischemia) was followed by 30 min of reperfusion. Immunoglobulin G liposomes (IgG-L) or CSIL was also infused at 5, 10, and 20 min of ischemia, reperfused, and then prepared for histochemical staining and electron microscopy. RESULTS: Recovery of left ventricular developed pressure (LVDP) of ischemic hearts treated with CSIL at 1 min of ischemia, assessed at 5 min of reperfusion (98 +/- 14%), was similar to that of sham-operated hearts (100%) but was significantly greater than that of placebo-treated hearts (12 +/- 7%, p = 0.01). The LVDP of hearts treated with CSIL at 5, 10, and 20 min was significantly greater than that with IgG-L at corresponding times (p < 0.03). Histochemical integrity and ultra-structural myocardial integrity were consistent with the functional data. CONCLUSIONS: Preservation of myocardial viability ex vivo was achieved with CSIL therapy. The extent of preservation is proportional to the time of initiation of therapy. Beneficial effects were observed even when CSIL therapy was initiated at 20 min of global ischemia. Therefore, delayed CSIL intervention after the onset of ischemia may augment preservation of myocardial viability during reperfusion therapy. PMID- 15120832 TI - Delayed response of insulin-stimulated fluorine-18 deoxyglucose uptake in glucose transporter-4-null mice hearts. AB - OBJECTIVES: We sought to evaluate the time course of insulin-stimulated myocardial glucose uptake (MGU) in mice that had undergone ablation of glucose transporter-4 (GLUT4). BACKGROUND: The relative importance of GLUT4, the most abundant insulin-responsive glucose transporter, to modulate myocardial glucose metabolism is not well defined. METHODS: Myocardial glucose uptake was assessed at various time points after glucose (1 mg/g) and insulin (8 mU/g) injection in GLUT4-null (G4N) (n = 48) and wild-type (WT) (n = 48) mice with (18)F-2-deoxy-2 fluoro-d-glucose (FDG) using in vivo positron emission tomography (PET), in vitro gamma-counter biodistribution, and isolated, perfused hearts. RESULTS: Baseline assessment with PET imaging showed comparable MGU in G4N (0.66 +/- 0.12) and WT (0.67 +/- 0.11, p = 0.70) mice. Early after insulin injection, WT mice demonstrated a 3.5-fold increase in MGU (2.45 +/- 0.45, p = 0.03), whereas G4N mice presented no increase (1.11 +/- 0.24, p = 0.28). At 60 min, MGU was comparable in G4N (3.19 +/- 0.60) and WT (2.66 +/- 0.47, p = 0.28) mice. In vitro gamma-counter biodistribution evaluation confirmed in G4N mice a lack of MGU increase early after insulin, but a slow response over 120 min. The isolated, perfused hearts of G4N mice during short-term (15 min) insulin stimulation displayed no increase in MGU (0.08 +/- 0.01 ml/g/min), whereas WT mice presented a threefold increase (0.22 +/- 0.01 ml/g/min, p < 0.01). With long-term (60 min) insulin stimulation, similar MGU was found in G4N (0.31 +/- 0.02 ml/g/min) and WT (0.33 +/- 0.04 ml/g per min, p = 0.04) mice. CONCLUSIONS: The G4N mice displayed an increase of MGU in response to insulin similar to that of controls, but with a markedly delayed time response. Our findings underscore the important role of GLUT4 in the rapid adaptive response of myocardial glucose metabolism. PMID- 15120833 TI - Osteopontin modulates angiotensin II-induced fibrosis in the intact murine heart. AB - OBJECTIVES: Osteopontin (OPN) is upregulated in left ventricular hypertrophy and is stimulated by angiotensin II (AngII). Our objective was to determine whether mice deficient in OPN would be protected from AngII-induced cardiac fibrosis. BACKGROUND: Interstitial fibrosis can lead to myocardial dysfunction and ultimately heart failure. Osteopontin activates integrins that regulate cell adhesion, migration, and growth, thus implicating OPN in the process of cardiac fibrosis. METHODS: Osteopontin null (OPN(-/-)) mice (n = 18) and wild-type controls (n = 20) were infused with AngII (2.5 or 3.0 microg/kg/min) for four days or three weeks via osmotic mini-pumps. Hearts were assessed morphometrically and histologically, including quantitative assessment of fibrosis via optical microscopic imaging analysis. Cardiac fibroblasts derived from these mice were evaluated for adhesion and proliferation. Cardiac transcript expression for cytokines, extracellular matrix (ECM), integrin, and atrial natriuretic peptide were assessed. RESULTS: Osteopontin(-/-) mice exhibited less cardiac fibrosis (0.7%) than wild-type mice (8.0%) (p < 0.01) and lowered heart/body weight ratios (0.10% vs. 0.23%) (p < 0.01) after three weeks of AngII infusion. Expression of transforming growth factor-beta, fibronectin, and collagen was not different between OPN(-/-) and wild-type mice, despite the decrease in ECM accumulation in the OPN(-/-) mice. Adhesion to ECM substrates decreased by 30% to 50% in cardiac fibroblasts of OPN(-/-) mice but was restored in OPN(-/-) cells by the addition of recombinant osteopontin. CONCLUSIONS: Osteopontin mediates cardiac fibrosis, probably through the modulation of cellular adhesion and proliferation. Because OPN is increased in cardiac hypertrophy and its lack attenuates fibrosis, understanding of OPN function is essential to extend our knowledge about molecular determinants of cardiac hypertrophy and failure. PMID- 15120834 TI - The year in cardiovascular surgery. PMID- 15120835 TI - Mode of initiation and ablation of ventricular fibrillation storms in patients with ischemic cardiomyopathy. AB - OBJECTIVES: We report on the initiation of ventricular fibrillation (VF) storm in patients with ischemic cardiomyopathy (ICM) and the results of targeted ablation to treat VF storm. BACKGROUND: Monomorphic premature ventricular contractions (PVCs) have been shown to initiate VF in patients without structural heart disease. METHODS: A total of 29 patients with ICM and documented VF initiation were identified. In 21 patients, VF storm was controlled with antiarrhythmic drugs and/or treatment of heart failure. Eight patients with VF (mean 52 +/- 25 episodes) refractory to medical management required ablation. All patients underwent three-dimensional electroanatomical mapping using CARTO (Biosense Webster Inc., Diamond Bar, California), and PVCs were mapped when present. Scarred areas were identified using voltage mapping. RESULTS: Monomorphic PVCs initiated VF in all 29 identified patients. Five of eight patients requiring ablation had frequent PVCs that allowed PVC mapping. The earliest activation site was consistently located in the scar border zone. The PVCs were always preceded by a Purkinje-like potential (PLP). Ablation was successfully performed at these sites. In three patients, infrequent PVCs prevented mapping, but PLPs were recorded around the scar border. Ablation targeting these potentials along the scar border was successfully performed. During follow-up (10 +/- 6 months), one patient had a single VF episode and another developed sustained, monomorphic ventricular tachycardia. There was no recurrence of VF storm. CONCLUSIONS: Ventricular fibrillation in ICM is triggered by monomorphic PVCs originating from the scar border zone with preceding PLPs; targeting these PVCs may prevent VF recurrence. In the absence of PVCs, both substrate mapping and ablation appear to be equally effective. PMID- 15120836 TI - President's page: Strengthening our house of cardiology. PMID- 15120837 TI - Diuretic use, progressive heart failure, and death in patients in SOLVD. PMID- 15120839 TI - Sympathetic nervous system activation in chagasic patients with congestive heart failure. PMID- 15120841 TI - Assessment of diastolic function using myocardial velocity gradient. PMID- 15120843 TI - Permanent occlusion of the middle cerebral artery upregulates expression of cytokines and neuronal nitric oxide synthase in the spinal cord and urinary bladder in the adult rat. AB - The expression pattern of proinflammatory cytokines, neuronal nitric oxide synthase (nNOS), substance P (SP) and calcitonin gene related peptide (CGRP) in the spinal cord and the bladder in response to permanent middle cerebral artery occlusion (MCAO) was investigated. In this connection, the gene expression of tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta) and interleukin-6 in the lumbosacral spinal cord and the bladder as determined by real-time polymerase chain reaction was upregulated. In the spinal cord, the immunoreactivity of TNF-alpha and IL-1beta was mainly localized in the ventral horn motoneurons contralateral to MCAO. In the bladder, TNF-alpha was mainly expressed in the inflammatory cells. The expression of nNOS immunoreactivity as well as nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) staining in the spinal cord and bladder was also markedly increased in response to MCAO. Furthermore, the temporal and spatial expression of nNOS paralleled that of TNF alpha and IL-1beta in the spinal cord. On the other hand, there was no noticeable change in gene expression and immunoreactivity of SP and CGRP. The present results have shown that cytokines and nNOS expression are elevated in areas far removed from the primary site of ischemic infarct, namely, the lumbosacral spinal cord and bladder. This together with some neuronal deaths maybe linked to the dysfunction of the latter in a clinical stroke. On the other hand, the apparent lack of SP and CGRP changes following MCAO suggests that the two neurotransmitters are not directly involved. PMID- 15120844 TI - Neuronal localization of the 25-kDa specific thiamine triphosphatase in rodent brain. AB - Thiamine triphosphate (ThTP) is found in small amounts in most organisms from bacteria to mammals, but little is known about its physiological role. In vertebrate tissues, ThTP may act as a phosphate donor for the phosphorylation of certain proteins; this may be part of a new signal transduction pathway. We have recently characterized a highly specific 25-kDa thiamine triphosphatase (ThTPase) that is expressed in most mammalian tissues. The role of this enzyme may be the control of intracellular concentrations of ThTP. As the latter has been considered to be a neuroactive form of thiamine, we have studied the distribution of ThTPase mRNA and protein in rodent brain using in situ hybridization and immunohistochemistry. With both methods, we found the strongest staining in hippocampal pyramidal neurons, as well as cerebellar granule cells and Purkinje cells. Some interneurons were also labeled and many ThTPase mRNA-positive and immunoreactive cells were distributed throughout cerebral cortical gray matter and the thalamus. White matter was not significantly labeled. ThTPase immunoreactivity seems to be located mainly in the cytoplasm of neuronal perikarya. Immunocytochemical data using dissociated cultured cells from hippocampal and cerebellum showed that the staining was more intense in neurons than in astrocytes. The protein was rather uniformly located in the perikarya and dendrites, suggesting that ThTP and ThTPase may play a general role in neuronal metabolism rather than a specific role in excitability. There was no apparent correlation between ThTPase expression and selective vulnerability of certain brain regions to thiamine deficiency. PMID- 15120845 TI - A Ca2+-independent slow afterhyperpolarization in substantia nigra compacta neurons. AB - The discharge properties of dopaminergic neurons in substantia nigra are influenced by slow adaptive responses, which have not been fully identified. The present study describes, in a slice preparation from the rat, a complex afterhyperpolarization (AHP), elicited by action potential trains. The AHP could be subdivided into a fast component (AHP(f)), which was generated near action potential threshold, relaxed within approximately 1 s, and had highest amplitude when evoked by short-lasting (0.1 s) depolarizations, and a slow component (AHP(s)), which lasted several seconds, was evoked from subthreshold potentials, and required prolonged depolarizing stimuli (>0.1 s). A large proportion of the AHP(f) was sensitive to (i) 0.1 microM apamin, (ii) the Ca(2+) antagonists, Cd(2+) (0.2 mM) and Ni(2+) (0.3 mM), (iii) low (0.2 mM) extracellular Ca(2+) concentration, and (iv), Ca(2+) chelation with intracellular EGTA. The AHP(s) was resistant to the above treatments, and it was insensitive to 25 microM dantrolene or prolonged exposure to 1 microM thapsigargin. The reversal potential of the AHP(s) (-97 mV) was close to the K(+) equilibrium potential. It was significantly inhibited by 5 mM 4-aminopyridine, 5 microM haloperidol, 10 microM terfenadine, or high extracellular Mg(2+) (10 mM), but not by 30 mM tetraethylammonium chloride, 50 microM carbachol, 0.5 microM glipizide, 2 microM (-)sulpiride, 100 microM N-allyl-normetazocine, or 100 microM pentazocine. Haloperidol reduced the post-stimulus inhibitory period seen during spontaneous discharge, but had no detectable effect on spike frequency adaptation. It is concluded that the SK-type Ca(2+)-activated K(+) channels underlies a major component of the AHP(f), whereas the AHP(s) is Ca(2+)-independent and relies, in part, on a voltage-dependent K(+) current with properties resembling the ether-a-go-go-related gene K(+) channel. The latter component exerts a slow, spike-independent, inhibitory influence on repetitive discharge and contributes to the prolonged decrease in excitability following sustained depolarizing stimuli. PMID- 15120846 TI - Neurotensin depolarizes globus pallidus neurons in rats via neurotensin type-1 receptor. AB - The globus pallidus is a major component in the indirect pathway of the basal ganglia. There is evidence that neurotensin receptors exist in this nucleus. To determine the electrophysiological effects of neurotensin on pallidal neurons, whole-cell patch-clamp recordings were performed in the acutely prepared brain slices. Under current-clamp recordings, neurotensin at 1 microM depolarized pallidal neurons. Voltage-clamp recordings also showed an inward current induced by neurotensin. The depolarizing effect of neurotensin could be mimicked by the C terminal fragment, neurotensin (8-13), but not by the N-terminal fragment, neurotensin (1-8). Both SR 142948A, a non-selective neurotensin receptor type-1 and type-2 antagonist, and SR 48692, a selective type-1 receptor antagonist, blocked the depolarizing effect of neurotensin, and which themselves had no effect on membrane potential. Thus, neurotensin type-1 receptors appear to mediate the effect of neurotensin. The depolarization evoked by neurotensin persisted in the presence of tetrodotoxin, ionotropic and metabotropic glutamate and GABA receptor antagonists, indicating that neurotensin excited the pallidal neurons by activating the receptor expressed on the neurons recorded. Current voltage relationship revealed that both the suppression of a potassium conductance and the activation of a cationic conductance are involved in the neurotensin-induced depolarization. Based on the action of neurotensin in the globus pallidus we hypothesize that alterations of the striatopallidal neurotensin system contribute to symptoms of basal ganglia motor disorders. PMID- 15120847 TI - Proton conductance of human transient receptor potential-vanilloid type-1 expressed in oocytes of Xenopus laevis and in Chinese hamster ovary cells. AB - Transient receptor potential-vanilloid type-1 (TRPV1) is a ligand-gated cation channel with preference for divalent cations, especially Ca(2+) (sequence of conductances: Ca(2+)>Mg(2+)>Na(+) approximately/= K(+) approximately/= Cs(+)). In the present study, the two-electrode voltage-clamp technique was used on oocytes of Xenopus laevis expressing TRPV1 to evaluate whether human TRPV1 also conducts protons. In medium devoid of K(+), Na(+), Mg(2+), and Ca(2+), capsaicin 1 microM induced a significant inward current (62% of the current in physiological medium). The effects of capsaicin were abolished in the presence of capsazepine 3 microM. The capsaicin-induced currents in medium devoid of Na(+), K(+), Mg(2+), and Ca(2+) were dependent on pH, causing larger inward currents and less negative reversal potentials at low pH and vice versa. The same current was also demonstrated in Chinese hamster ovary cells expressing human TRPV1. We conclude that TRPV1 conducts protons, in addition to Na(+), K(+), Mg(2+), and Ca(2+). The proton conductance may help to initiate action potentials and to translocate H(+) dependent on TRPV1 activation and membrane potential. PMID- 15120848 TI - Characterization of extracellular accumulation of Zn2+ during ischemia and reperfusion of hippocampus slices in rat. AB - The mammalian CNS contains an abundance of chelatable zinc that is sequestered in the vesicles of glutamatergic presynaptic terminals and co-released with glutamate. Considerable Zn(2+) is also released during cerebral ischemia and reperfusion (I/R) although the mechanism of this release has not been elucidated. We report here the real time observation of increase of the concentration of extracellular Zn(2+) ([Zn(2+)](o)), accompanied by a rapid increase of intracellular free Zn(2+)concentration, in the areas of dentate gyrus (DG), CA1 and CA3 in acute rat hippocampus slices during ischemia simulated by deprivation of oxygen and glucose (OGD) followed by reperfusion with normal artificial cerebrospinal fluid. A brief period of OGD caused a sustained increase of [Zn(2+)](o). Subsequent reperfusion with oxygenated medium containing glucose resulted in a further increase of [Zn(2+)](o). Longer periods of OGD caused greater increases of [Zn(2+)](o,) and subsequent reperfusion caused still further increases of [Zn(2+)](o,) regardless of OGD duration. The Zn(2+) chelator CaEDTA (10 mM) significantly reduced the increase of [Zn(2+)] induced by OGD and reperfusion. Significant regional differences of [Zn(2+)](o) over the areas of the DG, CA1 and CA3 were not observed during I/R. Neither sodium channel blockade by tetrodotoxin (2 microM), perfusion with nominally calcium-free medium nor anatomical disassociation of the DG, CA1 and CA3 regions from one another by lesioning affected the increase of [Zn(2+)](o). The non-specific nitric oxide synthase (NOS) inhibitor, Nomega-nitro-l-arginine methyl ester (1 mM), however, blocked the increase of [Zn(2+)](o) during ischemia and reperfusion. The data indicate the important role of NO in causing the release of Zn(2+) during I/R and suggest that NOS inhibitors may be used to reduce Zn(2+)-induced neuronal injury. PMID- 15120849 TI - Intracellular spermine decreases open probability of N-methyl-D-aspartate receptor channels. AB - Spermine and related polyamines have been shown to be endogenous regulators of several ion channel types including ionotropic glutamate receptors. The effect of spermine on N-methyl-d-aspartate (NMDA) receptors in cultured rat hippocampal neurons was studied using single-channel and whole-cell patch clamp recordings. Intracellular spermine resulted in the dose-dependent inhibition of NMDA-induced responses. Spermine reversibly inhibited the single NMDA receptor channel activity in inside-out patches suggesting a membrane-delimited mechanism of action. Open probability of NMDA receptor channels was decreased in a dose dependent manner. Mechanism of spermine-induced inhibition of NMDA receptor was different from that of intracellular Ca(2+)-induced NMDA receptor inactivation. Both pharmacological studies and single channel analysis indicate that in contrast to the effect of extracellular spermine the intracellular spermine effect is not dependent on the NMDA receptor subunit composition. We propose that intracellular spermine has a direct inhibitory effect on NMDA receptors that is different from calcium-induced NMDA receptor inactivation and spermine-induced voltage-dependent inhibition of AMPA/kainate receptors. Spermine-induced tonic change in the open probability of NMDA receptor channels may play a role in mechanisms underlying short-term changes in the synaptic efficacy. PMID- 15120850 TI - Basal forebrain neurons modulate the synthesis and expression of neuropeptides in the rat suprachiasmatic nucleus. AB - We tested the hypothesis that efferents from the nucleus basalis magnocellularis (NBM) play a direct role in the regulation of neuropeptide synthesis and expression by neurons of the rat suprachiasmatic nucleus (SCN). Adult male rats in which the NBM was destroyed with quinolinic acid, either unilaterally or bilaterally, were compared with rats injected with physiological saline and with control rats. The estimators used to assess the effects of cholinergic deafferentation on the neuroanatomy and neurochemistry of the SCN were the total number of SCN neurons, the total number and somatic size of SCN neurons producing vasopressin (VP) and vasoactive intestinal polypeptide (VIP), and the respective mRNA levels. Bilateral destruction of the NBM did not produce cell death in the SCN, but caused a marked reduction in the number and somatic size of SCN neurons expressing VP and VIP, and in the mRNA levels of these peptides. The decrease in the number of VP- and VIP-producing neurons provoked by unilateral lesions was less striking than that resulting from bilateral lesions. It was, however, statistically significant in the ipsilateral hemisphere, but not in the contralateral hemisphere. The results show that the reduction of cholinergic inputs to the SCN impairs the synthesis, and thereby decreases the expression of neuropeptides by SCN neurons, and that the extent of the decline correlates with the amount of cholinergic afferents destroyed. This supports the notion that acetylcholine plays an important, and direct role in the regulation of the metabolic activity of SCN neurons. PMID- 15120851 TI - Effects of adenoviral-mediated gene transfer of interleukin-10, interleukin-4, and transforming growth factor-beta on the survival of axotomized retinal ganglion cells. AB - Nitric oxide, synthesized by reactive microglia and astrocytes has been implicated in promoting neuronal degeneration observed in many diseases and insults of the central nervous system. We have recently shown that inducible nitric oxide synthase is expressed by retinal glial cells following optic nerve transection and that inhibition of nitric oxide synthesis enhances the survival of injured retinal ganglion cells. Anti-inflammatory cytokines including interleukin-10 (IL-10), interleukin-4 (IL-4), and transforming growth factor-beta (TGF-beta) have been shown to prevent inducible nitric oxide synthase expression, and inhibit nitric oxide synthesis by microglia and astrocytes in culture. In the present study, we examined the effects of adenoviral mediated gene transfer of anti-inflammatory cytokines on the survival of axotomized retinal ganglion cells. Intraocular administration of adenoviral vectors encoding interleukin-10 (Ad.IL 10) and interleukin-4 (Ad.IL-4) enhanced the survival of axotomized retinal ganglion cells at 14 days after axotomy. Adenoviral vectors encoding TGF-beta (Ad.TGF-beta) had no effect on retinal ganglion cell survival. Separate animals were pretreated by injection of Ad.IL-10 or Ad.IL-4 into the superior colliculus (s.c.), the major target of ganglion cells, 7 days prior to axotomy. S.c. administration of Ad.IL-10 or Ad.IL-4 significantly increased ganglion cell survival compared with intraocular injection. IL-10 and IL-4 gene transfer also reduced the density of infiltrating ED1 positive monocytes in the nerve fiber layer at 14 days postaxotomy. Ad.TGF-beta increased the density of ED1 positive monocytes infiltrating the nerve fiber layer after axotomy. Vectors encoding IL 10 or IL-4 also decreased nitrotyrosine immunoreactivity in the inner retina at 7 days postaxotomy, suggesting that these cytokines protect retinal ganglion cells from peroxynitrite formation that results from nitric oxide synthesis by activated glial cells. The present study has implications for the treatment of CNS injury and diseases that involve reactive microglia and astrocytes. Our results suggest that interleukin-10 and interleukin-4 may help prevent neurodegeneration caused by the activation of glial cells after CNS injury. PMID- 15120852 TI - Altered blood-brain barrier development in dystrophic MDX mice. AB - In order to ascertain whether the alterations of the blood-brain barrier (BBB) seen in adult dystrophic mdx-mice [Glia 42 (2003) 235], a human model of Duchenne muscular dystrophy (DMD), are developmentally established and correlated with other dystrophin isoforms which are localized at the glial-vascular interface, we used immunocytochemistry to investigate the expression of dystrophin isoforms (Dp71) during BBB development in mdx fetuses and in adult mice. Parallelly, we used Western blot, immunocytochemistry and immunogold electron microscopy to analyze the expression of the zonula occludens (ZO-1), aquaporin-4 (AQP4) and glial fibrillary acidic (GFAP) proteins as endothelial and glial markers, and we evaluated the integrity of the mdx BBB by means of intravascular injection of horseradish peroxidase (HRP). The results show reduced dystrophin isoforms (Dp71) in the mdx mouse compared with the control, starting from early embryonic life. Endothelial ZO-1 expression was reduced, and the tight junctions were altered and unlabeled. AQP4 and GFAP glial proteins in mdx mice also showed modifications in developmental expression, the glial vascular processes being only lightly AQP4- and GFAP-labeled compared with the controls. Confocal microscopy and HRP assays confirmed the alteration in vessel glial investment, GFAP perivascular endfoot reactivity being strongly reduced and BBB permeability increasing. These results demonstrate that a reduction in dystrophin isoforms (Dp71) at glial endfeet leads to an altered development of the BBB, whose no-closure might contribute to the neurological dysfunctions associated with DMD. PMID- 15120853 TI - Effect of neuronal nitric oxide synthase inhibition on CA2+/calmodulin kinase kinase and CA2+/calmodulin kinase IV activity during hypoxia in cortical nuclei of newborn piglets. AB - The present study tests the hypothesis that cerebral tissue hypoxia results in increased Ca(2+)/calmodulin (CaM) kinase kinase activity and that the administration of nitric oxide synthase inhibitors (N-nitro-l-arginine [NNLA], or 7-nitroindazole sodium [7-NINA]) prior to the onset of hypoxia will prevent the hypoxia-induced increase in the enzyme activity. To test this hypothesis, CaM kinase kinase and CaM kinase IV activities were determined in normoxic, hypoxic, NNLA-treated hypoxic, and 7-NINA-treated hypoxic piglets. Hypoxia was induced (FiO(2)=0.05-0.08x1 h) and confirmed biochemically by tissue levels of ATP and phosphocreatine. CaM kinase kinase activity was determined in a medium containing protein kinase and phosphatase inhibitors, calmodulin, and a specifically designed CaM kinase kinase target peptide. CaM kinase IV activity was determined by (33)P-incorporation into syntide-2 in a buffer containing protein kinase and phosphatase inhibitors. Compared with normoxic animals, ATP and phosphocreatine levels were significantly lower in all hypoxic piglets whether or not pretreated with nitric oxide synthase inhibitors. There was a significant difference among CaM kinase kinase activity (pmol/mg protein/min) in normoxic (76.84+/-14.1), hypoxic (138.86+/-18.2, P<0.05 vs normoxia), NNLA-pretreated hypoxic (91.34+/ 19.3; P=NS vs normoxia, P<0.05 vs hypoxia) and 7-NINA-pretreated hypoxic animals (100.12+/-23.3; P=NS vs normoxia, P<0.05 vs hypoxia). There was a significant difference among CaM kinase IV activity (pmol/mg protein/min) in normoxia (1270.80+/-126.1), hypoxia (2680.80+/-136.7; P<0.05 vs normoxia), NNLA-pretreated hypoxia (1666.00+/-154.8; P<0.05 vs normoxia, P<0.05 vs hypoxia), and 7-NINA pretreated hypoxic (1712.9+/-231.5; P=NS vs normoxia, P<0.05 vs hypoxia). We conclude that the hypoxia-induced increase in CaM kinase kinase and CaM kinase IV activity is mediated by neuronal NOS-derived NO. PMID- 15120854 TI - Neonatal manipulations of oxytocin alter expression of oxytocin and vasopressin immunoreactive cells in the paraventricular nucleus of the hypothalamus in a gender-specific manner. AB - Early postnatal manipulations of oxytocin have long-term behavioral and physiological consequences; the present study examined the hypothesis that oxytocin or its absence influences the subsequent expression of either oxytocin or arginine vasopressin in the CNS. On postnatal day 1 female and male prairie voles (Microtus ochrogaster) received a single i.p. injection of oxytocin (3 microg), oxytocin antagonist (0.3 microg), or 50 microl of isotonic saline or were only handled. On postnatal days 1, 8 and 21, brains were fixed, sectioned and stained for oxytocin or vasopressin immunoreactivity and analyzed as a function of age, treatment and sex. Both oxytocin and vasopressin immunoreactivity were observed on day 1 in the supraoptic and paraventricular nuclei (PVN) of the hypothalamus. Numbers of oxytocin and vasopressin neurons increased with age in both nuclei. Females treated on postnatal day 1 with oxytocin or oxytocin antagonist displayed a significant increase in oxytocin immunoreactivity on day 21 in the PVN. In contrast, males treated with antagonist tended to have decreased vasopressin immunoreactivity in the same region. These results revealed that the effects of neonatal manipulation of oxytocin are age dependent, site-specific and sexually dimorphic. The long-lasting effects of neonatal exposure to exogenous oxytocin and oxytocin antagonist indicate a role for oxytocin in the development of the CNS during the neonatal period, affecting the development of the oxytocinergic system in females and the vasopressinergic system in males. The developmental effects observed suggest one possible mechanism by which neonatal exposure to oxytocin or neonatal inhibition of endogenous oxytocin produces long-lasting behavioral and physiological alterations and could play a role in the development of male- and female-typical behavior. PMID- 15120855 TI - Nitric oxide is involved in nicotine-induced burst firing of rat ventral tegmental area dopamine neurons. AB - In the present study, using single cell recordings in vivo and intracellular recordings in vitro from midbrain slices, the role of N-methyl-d-aspartate (NMDA) receptor signaling on firing activity in ventral tegmental area dopamine neurons elicited by nicotine was investigated in the rat. In accordance with previous studies, systemic nicotine (0.5 mg/kg s.c.) increased both firing rate and burst firing of dopamine neurons in vivo, and bath-applied nicotine (10 microM) increased firing rate in vitro. The competitive NMDA receptor antagonist CGP39551 (2.5 mg/kg i.p.) inhibited nicotine's effects on burst firing and also attenuated the nicotine-induced increase in firing rate. Moreover, although the nitric oxide (NO)-synthase inhibitor N-nitro-l-arginine-methyl-ester (l-NAME; 5.0 mg/kg i.p.) had no effect on cell firing by itself, it prevented the response to nicotine in vivo. In contrast, l-NAME (100 microM) did not influence nicotine's effect on dopamine cell firing in vitro, suggesting that the effect of l-NAME seen in vivo is dependent on presynaptic afferent input. The present study confirms previous results suggesting that the effect of systemically administered nicotine is in part presynaptic and mediated via NMDA receptors. The data also indicate that NO plays an important role in the previously demonstrated, indirect, glutamate mediated excitation of these neurons by nicotine. By inference, our results provide additional support for the involvement of NO in nicotine dependence. PMID- 15120856 TI - Succinate increases neuronal post-synaptic excitatory potentials in vitro and induces convulsive behavior through N-methyl-d-aspartate-mediated mechanisms. AB - Succinate is a dicarboxylic acid that accumulates due to succinate dehydrogenase inhibition by malonate and methylmalonate exposure. These neurotoxins cause increased excitability and excitotoxic damage, which can be prevented by administering high amounts of succinate. In the present study we investigated whether succinate alters hippocampal field excitatory post-synaptic potentials. Bath application of succinate at intermediate concentrations (0.3-1 mM) increased the slope of field excitatory post-synaptic potentials in hippocampal slices, and at high concentrations (above 1 mM) did not alter or decrease field excitatory post-synaptic potentials slope. Succinate-induced enhancement of field excitatory post-synaptic potentials slope was abolished by the addition of d-2-amino-5 phosphonovaleric acid (50 microM) to the perfusate, supporting the involvement of N-methyl-d-aspartate receptors in the excitatory effect of this organic acid. Accordingly, succinate (0.8-7.5 micromol) i.c.v. administration caused dose dependent convulsive behavior in mice. The i.c.v. co-administration of MK-801 (7 nmol) fully prevented succinate-induced convulsions, further suggesting the involvement of N-methyl-d-aspartate receptors in the convulsant action of succinate. Our data indicate that accumulation of moderate amounts of succinate may contribute to the excitotoxicity induced by succinate dehydrogenase inhibitors, through the activation of N-methyl-d-aspartate receptors. PMID- 15120857 TI - Expression of adrenomedullin and proadrenomedullin N-terminal 20 peptide in PC12 cells after exposure to nerve growth factor. AB - Adrenomedullin (AM) and proadrenomedullin N-terminal 20 peptide (PAMP) are multi functional peptides derived from the same precursor, proadrenomedullin. We have studied the regulatory mechanism of expression of these peptides during neuronal differentiation of rat pheochromocytoma PC12 cells by nerve growth factor (NGF). The cellular levels of the peptides increased slightly, and then progressively decreased below the control by NGF. Immunoreactive (ir)-AM in the medium was transiently increased by NGF. Cytochemical staining showed that ir-AM and ir-PAMP were abundantly present in cytoplasm in the undifferentiated cells, and were decreased during culture with NGF. There was no preferential localization of ir AM or ir-PAMP in neurites in comparison with in cytoplasm in the differentiated cells. Northern blot analysis showed that mRNA encoding these peptides, as detected as a band of 1.6 kb, increased more than three-fold at 1 h after the addition of NGF and then progressively decreased to one fifth of the control during 72 h. Degradation rate of the mRNA was slowed by NGF even when mRNA level is decreased after 72 h of NGF treatment. The transcription rate of their gene increased transiently and then decreased by the long-term treatment with NGF. These results demonstrate that expression of AM and PAMP is regulated by NGF along with time-dependent differentiation: AM gene transcription is transiently activated by NGF, whereas it was suppressed during neuronal differentiation of the cells. PMID- 15120858 TI - Acute and delayed restraint stress-induced changes in nitric oxide producing neurons in limbic regions. AB - RATIONALE: Microinjection into the dentate gyrus of the hippocampus of N(omega) nitro-l-arginine methyl ester hydrochloride (l-NAME), a nitric oxide synthase (NOS) inhibitor, induces antinociceptive effect 5 days after a single restraint episode. The mechanisms of this stress-antinociceptive modulatory effect have not been investigated but may involve plastic changes in the hippocampal formation (HF). OBJECTIVE: The objective of the present study was to investigate possible mechanisms of the stress-modulating effect on antinociception induced by NOS inhibition in the hippocampus. We analyzed the effects of restraint stress on neuronal NOS (nNOS) expression and nicotinamide adenine dinucleotide phosphate diaphorase histochemical activity (NADPH-d) in the HF and related brain regions. METHODS: Male Wistar rats (n=6-11/group) were submitted to a single (acute stress) or repeated (5 days) episodes of 2-h restraint. Control animals remained in their home cages being all animals daily handled during this period. In the fifth day, animals received unilateral microinjection of l-NAME (150 nmol/0.2 microl) or saline (control) into the dentate gyrus of the dorsal hippocampus (DG). Immediately before and after drug microinjection tail-flick reflex latency or hotplate licking reaction was measured. Animals were killed i. immediately; ii. 5 days after acute stress; or iii. after repeated stress. NADPH-d and nNOS expression were quantified in the HF, caudate-putamen, secondary somatosensorial, entorhinal and piriform cortices and amygdaloid complex. RESULTS: Five days after one or five restraint episodes l-NAME microinjection into the DG elicited antinociceptive effect (analysis of variance [ANOVA], P<0.05). Acute restraint stress induced a significant increase in the density of neurons expressing NADPH d and nNOS in the amygdaloid nuclei. nNOS expression increased also in the DG and piriform cortex. Five days after a single or repeated restraint stress there was an additional increase in NADPH-d- and nNOS-positive neurons in CA1, CA3, and entorhinal cortex. No changes were seen in non-limbic regions such as the caudate putamen and secondary somatosensorial cortex. CONCLUSION: The results confirm that the dorsal hippocampus participates in the modulation of stress consequences. They also show that a single stress episode causes acute changes in nitric oxide system in the amygdala complex and delayed modifications in the HF. The delayed (5 days) antinociceptive effect of NOS inhibition in the HF after a single restraint episode suggests that those latter modifications may have functional consequences. It remains to be tested if the acute amygdala and delayed hippocampal changes are causally related. PMID- 15120859 TI - Opioidergic modulation of excitability of rat trigeminal root ganglion neuron projections to the superficial layer of cervical dorsal horn. AB - The aim of the present study was to investigate the effect of a micro-opioid receptor agonist DAMGO (Tyr-d-Ala-Gly-NMe-Phe-Gly-ol) on the excitability of trigeminal root ganglion (TRG) neurons, projecting onto the superficial layer of the cervical dorsal horn, by using the perforated-patch technique and to determine whether TRG neurons show the expression of mRNA or functional protein for micro-opioid receptors by using reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. TRG neurons projecting onto the superficial layer of the cervical dorsal horn were retrogradely labeled with Fluorogold (FG). The cell diameter of FG-labeled TRG neurons was small (<30 microm). Under voltage-clamp (V(h)=-60 mV), voltage-dependent K(+) currents were recorded in the TRG neurons and isolated by blocking Na(+) and Ca(2+) currents with appropriate ion replacement. Separation of the K(+) current components was achieved by the response to variation in the conditioning voltage. Two distinct K(+) current components, a transient (I(A)) and sustained (I(K)), were identified. DAMGO significantly increased I(A) by 57% (20 microM) and in a dose dependent manner (1-50 microM). Similarly, I(K) was also enhanced by DAMGO administration (42%, 20 microM). The augmentation of both I(A) and I(K) was antagonized by a micro-opioid receptor antagonist, CTOP (d-Phe-Cys-Thr-d-Trp-Orn Thr-Pen-Thr-NH(2)). Hyperpolarization of the membrane potential was elicited by DAMGO (20 microM) and the response was associated with a decrease in the input resistance. DAMGO induced hyperpolarization was blocked by CTOP. DAMGO-sensitive I(A) and I(K) currents were antagonized by K(+) channel blockers, 4-aminopyridine (4-AP) and tetraethylammonium (TEA). In the presence of both 4-AP and TEA, no significant changes in membrane potential induced by DAMGO application were observed. In the presence of BaCl(2), DAMGO evoked hyperpolarization with decreased resistance was observed. The firing rate of action potentials and the first spike duration induced by depolarizing step pulses were decreased in the presence of DAMGO. RT-PCR analysis demonstrated the expression of mRNA for micro opioid receptors in the trigeminal ganglia. The micro-opioid receptor immunoreactivity was expressed in the small diameter FG-labeled TRG neurons. These results suggest that the activation of micro-opioid receptors inhibits the excitability of rat small diameter TRG neurons projecting on the superficial layer of the cervical dorsal horn and this inhibition is mediated by potentiation of voltage-dependent K(+) currents. We therefore concluded that modulation of nociceptive transmission in the trigeminal system, resulting in the functional activation of micro-opioid receptors, occurs at the level of small TRG cell bodies and/or their primary afferent terminals, which contribute to opioid analgesia in the trigeminal pain. PMID- 15120860 TI - Central cholinergic functions in human amyloid precursor protein knock in/presenilin-1 transgenic mice. AB - Alzheimer's disease is characterized by amyloid peptide formation and deposition, neurofibrillary tangles, central cholinergic dysfunction, and dementia; however, the relationship between these parameters is not well understood. We studied the effect of amyloid peptide formation and deposition on central cholinergic function in knock-in mice carrying the human amyloid precursor protein (APP) gene with the Swedish/London double mutation (APP-SL mice) which were crossbred with transgenic mice overexpressing normal (PS1wt) or mutated (M146L; PS1mut) human presenilin-1. APP-SLxPS1mut mice had increased levels of Abeta peptides at 10 months of age and amyloid plaques at 14 months of age while APP-SLxPS1wt mice did not have increased peptide levels and did not develop amyloid plaques. We used microdialysis in 15-27 months old mice to compare hippocampal acetylcholine (ACh) levels in the two mouse lines and found that extracellular ACh levels were slightly but significantly reduced in the APP-SLxPS1mut mice (-26%; P=0.044). Exploratory activity in the open field increased hippocampal ACh release by two fold in both mouse lines; total and relative increases were not significantly different for the two strains under study. Similarly, infusion of scopolamine (1 microM) increased hippocampal ACh release to a similar extent (3-5-fold) in both groups. High-affinity choline uptake, a measure of the ACh turnover rate, was identical in both mouse lines. Neurons expressing choline acetyltransferase were increased in the septum of APP-SLxPS1mut mice (+26%; P=0.046). We conclude that amyloid peptide production causes a small decrease of extracellular ACh levels. The deposition of amyloid plaques, however, does not impair stimulated ACh release and proceeds without major changes of central cholinergic function. PMID- 15120861 TI - A novel modulatory mechanism of sodium currents: frequency-dependence without state-dependent binding. AB - We have previously found that the dopamine uptake inhibitor 1-(2-[bis(4 fluorophenyl)methoxy]ethyl)-4-(3-phenylpropyl)piperazine dihydrochloride (GBR 12909) inhibits neuronal sodium channels. The inhibition was profoundly dependent on the voltage protocol, suggesting that the effect is determined by the activity pattern of individual neurons. Our present study was aimed to understand more thoroughly the mechanism of this inhibition. The effect of GBR 12909 on sodium currents was investigated using whole-cell patch clamp recordings on cultured hippocampal neurons. Repetitive trains of depolarizations revealed two distinct components of inhibition: a frequency-dependent, transient and a frequency independent, sustained component. Frequency-dependent inhibition can reflect dynamic equilibrium of binding or gating. In order to decide which is the dominant mechanism in the case of GBR 12909, we studied the rates of association and dissociation. We found an unexpectedly fast rate of association (tau=819.2 ms) to resting ion channels kept at hyperpolarized membrane potential (-150 mV), while the rate of dissociation was too slow to explain recovery between trains of stimulation (tau=248 s). These data suggest that frequency-dependent inhibition cannot be explained by binding and unbinding, but rather it is due to conformational transitions of the liganded channel, which can only be explained if ligand binding is assumed to enhance slow inactivation. We studied, therefore, the rate of slow inactivation in the presence of different concentrations of GBR 12909. We have found that GBR 12909 accelerates slow inactivation substantially (time constants more than hundredfold lower at concentrations above 10 microM), causing the time range of slow inactivation to overlap with the time range of fast inactivation. Slow inactivation can even be the dominant process, especially during subthreshold depolarizations in the presence of >10 microM of GBR 12909. This mechanism of inhibition could provide a selective inhibition of neurons not only with high frequency bursting activity but also with moderately depolarized membrane potential. PMID- 15120863 TI - Y2 receptor expression and inhibition of voltage-dependent Ca2+ influx into rod bipolar cell terminals. AB - Neuropeptide Y (NPY) is a potent inhibitory neuropeptide expressed by amacrine cells in the rat retina. NPY modulates the release of multiple neurotransmitters in mammalian retina, yet the mechanisms mediating this regulation are not well defined. To further understand the action of NPY in the retina, Y receptor coupling to voltage-dependent Ca(2+) channels was investigated using Ca(2+) imaging with fura-2 AM to measure [Ca(2+)](i) increases in rod bipolar cell terminals. Y receptor expression was studied in rat retinal tissue with reverse transcription-polymerase chain reaction (RT-PCR). NPY inhibited the depolarization-evoked Ca(2+) influx into rod bipolar cell axon terminals and caused a dose-dependent reduction and an average maximal inhibition of 72% at 1 microM, which was reversed upon washout. K(+)-evoked Ca(2+) increases were also inhibited by the selective Y2 receptor agonists, C2-NPY and NPY(13-36), at concentrations of 1 microM, but not by the selective Y1 receptor agonist, [Leu(31)Pro(34)]NPY, selective Y4 receptor agonist, rPP, or the selective Y5 receptor agonist, [d-Trp32]-NPY. Y receptor expression was determined using RT PCR for all known Y receptor subtypes. Y2 receptor mRNA, as well as Y1, Y4, and Y5 receptor mRNAs, are present in the rat retina. Like the rod bipolar cell, other studies in central neurons have shown that the Y2 receptor is expressed predominantly as a presynaptic receptor and that it modulates transmitter release. Together, these findings suggest that NPY activates presynaptic Y2 receptors to inhibit voltage-dependent Ca(2+) influx into rod bipolar cell terminals, and establishes one mechanism by which NPY may reduce l-glutamate release from the rod bipolar cell synapse. PMID- 15120864 TI - Synaptic transmission of chaotic spike trains between primary afferent fiber and spinal dorsal horn neuron in the rat. AB - Primary sensory neurons can generate irregular burst firings in which the existence of significant deterministic behaviors of chaotic dynamics has been proved with nonlinear time series analysis. But how well the deterministic characteristics and neural information of presynaptic chaotic spike trains were transmitted into postsynaptic spike trains is still an open question. Here we investigated the synaptic transmission of chaotic spike trains between primary Adelta afferent fiber and spinal dorsal horn neuron. Two kinds of basic stimulus unit, brief burst and single pulse, were employed by us to comprise chaotic stimulus trains. For time series analysis, we defined "events" as the longest sequences of spikes with all interspike intervals less than or equal to a certain threshold and extracted the interevent intervals (IEIs) from spike trains. Return map analysis of the IEI series showed that the main temporal structure of chaotic input trains could be detected in postsynaptic output trains, especially under brief-burst stimulation. Using correlation dimension and nonlinear prediction methods, we found that synaptic transmission could influence the nonlinear characteristics of chaotic trains, such as fractal dimension and short-term predictability, with greater influence made under single-pulse stimulation. By calculating the mutual information between input and output trains, we found the information carried by presynaptic spike trains could not be completely transmitted at primary afferent synapses, and that brief bursts could more reliably transmit the information carried by chaotic input trains across synapses. These results indicate that although unreliability exists during synaptic transmission, the main deterministic characteristics of chaotic burst trains can be transmitted across primary afferent synapses. Moreover, brief bursts that come from the periphery can more reliably transmit neural information between primary afferent fibers and spinal dorsal horn neurons. PMID- 15120862 TI - Exogenous nerve growth factor attenuates opioid-induced inhibition of voltage activated Ba2+ currents in rat sensory neurons. AB - Nerve growth factor (NGF) promotes the survival of embryonic sensory neurons and maintains the phenotypic characteristics of primary nociceptive neurons postnatally. NGF also contributes to nociceptor activation and hyperalgesia during inflammatory pain states. The purpose of this study was to determine whether NGF might have an additional pronociceptive action by interfering with opioid-mediated analgesia in primary nociceptive neurons. Sensory neurons were isolated from the dorsal root ganglia of weanling rats and kept in standard culture conditions either with or without exogenous NGF (50 ng/ml). Currents through voltage-gated calcium channels were recorded from individual neurons using the whole cell patch clamp technique with Ba(2+) as the charge carrier (I(Ba)). The micro-opioid agonist fentanyl (1 microM) and the GABA(B) agonist baclofen (50 microM) were used to test G protein-dependent inhibition of I(Ba). Fentanyl inhibited I(Ba) by an average of 38+/-4% in untreated cells vs. 25+/-2% in NGF-treated cells (P<0.01). NGF had no effect on I(Ba) current magnitude or kinetics. The NGF-induced attenuation of opioid action was observed as early as 4 h after exposure, but was not seen when NGF was applied by bath perfusion for up to 40 min, suggesting that the effect was not mediated by a rapid phosphorylation event. The effect of NGF was prevented by K-252a (100 nM), an inhibitor of TrkA autophosphorylation. Baclofen-induced inhibition of I(Ba), on the other hand, was not affected by NGF treatment, suggesting that NGF modulation of opioid-mediated inhibition occurred upstream from the G protein. This was supported by the finding that GTP-gamma-S, an agonist independent G protein activator, inhibited I(Ba) similarly in both untreated and NGF treated cells. The results show that NGF selectively attenuated opioid-mediated inhibition of I(Ba) via TrkA receptor activation, possibly by altering opioid receptor function. PMID- 15120865 TI - Oestradiol dampens reflex-related activity of on- and off-cells in the rostral ventromedial medulla of female rats. AB - The present study was conducted to determine whether the ovarian steroid oestradiol alters the activity of nociceptive modulatory neurons in the rostral ventromedial medulla (RVM). Adult female rats were ovariectomized and implanted s.c. with an oestradiol-filled or placebo capsule. Sixteen to 37 days later, rats were anaesthetised for single unit recording from RVM neurons. On-cells were characterised by a burst of activity, and off-cells by a pause in activity immediately preceding reflexive withdrawal of the tail from 51 and 54 degrees C water. Although on- and off-cells were evident in both oestradiol- and placebo treated rats, the reflex-related on-cell burst and off-cell pause were dampened in oestradiol-treated rats. On-cells from oestradiol-treated rats had a mean activity burst of 9.1+/-2.2 Hz in the 2 s preceding the tail withdrawal reflex to 51 degrees C water, compared with 17.9+/-4.3 Hz for on-cells in placebo controls. Off-cell activity during the 2 s preceding tail withdrawal was 4.8+/-2.2 vs. 0.1+/-0.1 Hz in oestradiol vs. placebo-treated females, respectively. Similar changes in on- and off-cell activity occurred when the tail was placed in 54 degrees C water. The present data demonstrate that oestradiol constrains the magnitude of the shift in RVM on- and off-cell activity associated with nociceptive reflexes. PMID- 15120866 TI - Long-range temporal correlations in epileptogenic and non-epileptogenic human hippocampus. AB - Epileptogenic human hippocampus generates spontaneous energy fluctuations with a wide range of amplitude and temporal variation, which are often assumed to be entirely random. However, the temporal dynamics of these fluctuations are poorly understood, and the question of whether they exhibit persistent long-range temporal correlations (LRTC) remains unanswered. In this paper we use detrended fluctuation analysis (DFA) to show that the energy fluctuations in human hippocampus show LRTC with power-law scaling, and that these correlations differ between epileptogenic and non-epileptogenic hippocampus. The analysis shows that the energy fluctuations exhibit slower decay of the correlations in the epileptogenic hippocampus compared with the non-epileptogenic hippocampus. The DFA-derived scaling exponents demonstrate that there are LRTC of energy fluctuations in human hippocampus, and that the temporal persistence of energy fluctuations is characterized by a bias for large (small) energy fluctuations to be followed by large (small) energy fluctuations. Furthermore, we find that in the period of time leading up to seizures there is no change in the scaling exponents that characterize the LRTC of energy fluctuations. The fact that the LRTC of energy fluctuations do not change as seizures approach provides evidence that the local neuronal network dynamics do not change in the period before seizures, and that seizures in mesial temporal lobe epilepsy may be triggered by an influence that is external to the hippocampus. The presence of LRTC with power law scaling does not imply a specific mechanism, but the finding that temporal correlations decay more slowly in epileptogenic hippocampus provides electrophysiologic evidence that the underlying neuronal dynamics are different within the epileptogenic hippocampus compared with contralateral hippocampus. We briefly discuss possible neurobiological mechanisms for LRTC of the energy fluctuations in hippocampus. PMID- 15120867 TI - Fluctuations in somatosensory responsiveness and baseline firing rates of neurons in the lateral striatum of freely moving rats: effects of intranigral apomorphine. AB - Somatosensory responsiveness and baseline firing rates of 102 striatal neurons were studied in freely moving rats. For individual neurons, mean levels of responsiveness and baseline firing fluctuated unpredictably in direction and magnitude and independently of each other throughout an experiment. Following microinjections of apomorphine into the substantia nigra, which were used as a means of reducing nigral output activity, the magnitude of fluctuations in striatal somatosensory responsiveness significantly increased, while the magnitude of fluctuations in baseline firing was unaltered. The receptive zones of 54 neurons studied in control experiments remained stable, whereas receptive zones changed in 12 of 25 neurons studied after apomorphine microinjection. Normal nigrostriatal dopamine transmission appears to selectively restrict the magnitude of fluctuations in responsiveness of striatal neurons to corticostriatal synaptic input and may exert additional control over afferent projections from cutaneous receptive zones to these neurons. PMID- 15120868 TI - Risk-based maintenance of ethylene oxide production facilities. AB - This paper discusses a methodology for the design of an optimum inspection and maintenance program. The methodology, called risk-based maintenance (RBM) is based on integrating a reliability approach and a risk assessment strategy to obtain an optimum maintenance schedule. First, the likely equipment failure scenarios are formulated. Out of many likely failure scenarios, the ones, which are most probable, are subjected to a detailed study. Detailed consequence analysis is done for the selected scenarios. Subsequently, these failure scenarios are subjected to a fault tree analysis to determine their probabilities. Finally, risk is computed by combining the results of the consequence and the probability analyses. The calculated risk is compared against known acceptable criteria. The frequencies of the maintenance tasks are obtained by minimizing the estimated risk. A case study involving an ethylene oxide production facility is presented. Out of the five most hazardous units considered, the pipeline used for the transportation of the ethylene is found to have the highest risk. Using available failure data and a lognormal reliability distribution function human health risk factors are calculated. Both societal risk factors and individual risk factors exceeded the acceptable risk criteria. To determine an optimal maintenance interval, a reverse fault tree analysis was used. The maintenance interval was determined such that the original high risk is brought down to an acceptable level. A sensitivity analysis is also undertaken to study the impact of changing the distribution of the reliability model as well as the error in the distribution parameters on the maintenance interval. PMID- 15120869 TI - Phytotoxicity and heavy metals speciation of stabilised sewage sludges. AB - The presence of heavy metals in the sludges produced in wastewater treatment restricts plants growth and hence their use for agricultural purposes. This study looks at different types of sludges (aerobic, anaerobic, unstabilised and sludge from a waste stabilisation pond) and compares the distribution of the heavy metals that they contain according to the treatment that they have undergone. The sewage sludges were subjected to chemical characterisation and phytotoxicity testing (in absence of substrate) to provide a preliminary assessment of their suitability for land application. In addition, the total quantity of metals (Cd, Cr, Cu, Ca, K, Fe, Mg, Ni, Na, Pb and Zn) was determined. The Community Bureau of Reference (BCR) method for heavy metal speciation was followed. It was confirmed that the total concentration of heavy metals did not exceed the limits set out by European legislation and that the stabilisation treatment undergone by the sludges strongly influenced the heavy metal distribution and the phases to which they were associated. The sludge extracts did not exert any significant adverse effect on the relative seed germination (RSG) of barley (Hordeum vulgare L.) or cress (Lepidium sativum L.), although the reduction in germination index (GI) indicates that some characteristics existed did have an adverse effect on root growth. PMID- 15120870 TI - Kinetics and mechanism of chloride based chlorine dioxide generation process from acidic sodium chlorate. AB - The reaction between sodium chlorate and sodium chloride in presence of aqueous sulfuric acid is studied in a well stirred reactor at various temperatures and molar concentrations of chlorate, chloride and acid. The reaction rate is evaluated by analyzing the consumption of chlorate or chloride iodometrically or argentometrically. The rate law is established. Reaction has been found first and second order with respect to chlorate and chloride concentration, respectively. Reaction order is found about 13-14 with respect to molar concentration of sulfuric acid but it reduced to 2.4 when acidity function, h_, is substituted in place of molar concentration. The temperature dependence of the reaction is also investigated and pre-exponential Arrhenius parameter as well as activation energy are determined. It has been observed that ClO(2)/Cl(2) ratio is markedly affected by chlorate to chloride ratio. Reaction mechanism compatible with the reaction kinetics is proposed. PMID- 15120871 TI - Sorption of basic dyes from aqueous solution by activated sludge. AB - The adsorption of dyes in the solutions using activated sludge might be a promising approach in wastewater treatment units. The adsorption of Basic Red 18 and Basic Blue 9 from aqueous solution by dried activated sludge was investigated with in a batch system. The activated sludge had the highest dye uptake capacity, having the monolayer adsorption capacity 285.71 and 256.41 mg g(-1) for Basic Red 18 and Basic Blue 9, respectively, at pH value of 7.0 and 20 degrees C. Langmuir and Freundlich adsorption models were used for the mathematical description of the adsorption equilibrium and the equilibrium data fixed very well with both the Langmuir and Freundlich models. The R(L) values showed that, activated sludge was favorable for the adsorption of basic dyes. The suitability of the kinetic models for the adsorption of dyes on the activated sludge was also discussed. It was clear that the adsorption kinetics of dyes to dried activated sludge obeyed pseudo second-order adsorption kinetics. PMID- 15120872 TI - N2O removal in N2 or air by ArF excimer laser photolysis at atmospheric pressure. AB - A photochemical process is proposed as a new efficient N(2)O removal technique in N(2) or air at atmospheric pressure and room temperature without using any catalysts. N(2)O diluted in N(2) or air was decomposed into N(2), O(2), and NO by using a 193 nm ArF excimer laser. The maximum conversion of N(2)O in N(2)O/N(2) or N(2)O/N(2)/O(2) mixtures was 93% at a laser power of 136 mJ, a repetition frequency of 5 Hz, and an irradiation time of 30 min. The formation ratios of N(2):O(2):NO in N(2)O/N(2) and N(2)O/N(2)/O(2) mixtures were 64:31:5.1% and 60:27:13%, respectively. The decomposition mechanism of N(2)O under 193 nm photolysis was discussed by comparing experimental data with calculated model using known photochemical and gas kinetic data. PMID- 15120873 TI - Degradation products of the process of thermal recovery of copper from lamina scraps in lab-scale fluidized bed reactor. AB - This paper presents experimental results dealing with a process for recovering copper in the scrap composite materials issued from electronic laminas industry. This environment-friendly process consists in the thermal treatment of scrap in a fluidized bed whose particles fix the harmful gases emitted by the organic glue gasification. A series of experiments was carried out in a thermobalance coupled to FTIR spectrometer and GC/MS with small lamina samples. These experiments demonstrated the thermal behavior of scrap composite materials, and identified the major degradation reaction gases. A series of experiments was performed with bigger scrap samples hung in a laboratory-scale fluidized bed coupled to FTIR and MS, at 350 degrees C; the results confirmed those obtained in thermobalance. Experiments showed that a residence time lasting less than 5 min is sufficient to recover the metallic copper, and exhaust gases are not harmful. PMID- 15120874 TI - Predicting organic loading in natural water using spectral fluorescent signatures. AB - Spectral fluorescent signature (SFS) is a rapid, reagent free and inexpensive technique, which has great potential for environmental monitoring of aqueous systems, especially for predicting dissolved organic carbon (DOC) along natural waters. This technical note aimed to examine the possibility to use SFS associated with partial least squares regression (PLS) to assess the organic loading in natural water. A model was built using samples of water collected between October 1999 and February 2002 on the Passaic River at Little Falls, NJ, USA. A correlation was established between measured DOC, SFS, and the corresponding daily registered flow from United States Geological Survey (USGS) New Jersey's streamflow database. The methodology presented herein looks promising in making use of the significant organic characteristics information contained in a SFS for application and use in spatial and temporal water quality management and treatment. PMID- 15120875 TI - Metal elution from Ni- and Fe-based alloy reactors under hydrothermal conditions. AB - Elution of metals from Ni- and Fe-based alloy (i.e. Inconel 625 and SUS 316) under hydrothermal conditions was investigated. Results showed that metals could be eluted even in a short contact time. At subcritical conditions, a significant amount of Cr was extracted from SUS 316, while only traces of Ni, Fe, Mo, and Mn were eluted. In contrast, Ni was removed in significant amounts compared to Cr when Inconel 625 was tested. Several factors including temperature and contact time were found to affect elution behavior. The presence of air in the fluid even promoted elution under subcritical conditions. PMID- 15120876 TI - Obstructive sleep apnoea, metabolic syndrome, and cardiovascular outcomes. PMID- 15120877 TI - Statins in patients at high risk of cardiovascular disease presenting with peripheral artery disease. PMID- 15120878 TI - Assessment of reperfusion-induced myocardial injury by echocardiography. PMID- 15120879 TI - Treating low HDL-cholesterol in normocholesterolaemic patients with coronary disease: statins, fibrates or horses for courses? PMID- 15120880 TI - Long-term clopidogrel therapy after percutaneous coronary intervention in PCI CURE and CREDO: the "Emperor's New Clothes" revisited. PMID- 15120881 TI - Fractional flow reserve and complex coronary pathologic conditions. AB - Fractional flow reserve (FFR) is a well-validated index for assessing the physiologic significance of a coronary stenosis in most clinical conditions encountered in today's catheterisation laboratory. The aim of this paper is to provide a short overview of the theoretical background of the coronary pressure derived FFR index and its clinical applicability in guiding complex coronary intervention procedures. PMID- 15120882 TI - Benefits of obstructive sleep apnoea treatment in coronary artery disease: a long term follow-up study. AB - AIM: The aim of this long-term prospective study was to evaluate the effect of treating obstructive sleep apnoea (OSA) on the rate of cardiovascular events in coronary artery disease (CAD). METHODS AND RESULTS: We prospectively studied 54 patients (mean age 57.3 +/- 10.1 years) with both CAD (> or = 70% coronary artery stenosis) and OSA (apnoea-hypopnoea index > or = 15). In 25 patients, OSA was treated with continuous positive airway pressure (n=21) or upper airway surgery (n=4); the remaining 29 patients declined treatment for their OSA. The median follow-up was 86.5 +/- 39 months. The two groups were similar at baseline in age, body mass index, smoking history, hypertension, hypercholesterolaemia, diabetes mellitus, number of diseased vessels, left ventricular ejection fraction, and CAD therapy. Treatment of risk factors other than OSA was similar in the two groups. The endpoint (a composite of cardiovascular death, acute coronary syndrome, hospitalisation for heart failure, or need for coronary revascularisation) was reached in 6 (6/25, 24%) and 17 (17/29, 58%) patients with and without OSA treatment, respectively (P<0.01). OSA treatment significantly reduced the risk of occurrence of the composite endpoint (hazard ratio 0.24; 95% confidence interval, 0.09-0.62; p<0.01) and of each of its components. CONCLUSIONS: Our data indicate that the treatment of OSA in CAD patients is associated with a decrease in the occurrence of new cardiovascular events, and an increase in the time to such events. PMID- 15120883 TI - Obstructive sleep apnoea is independently associated with an increased prevalence of metabolic syndrome. AB - AIMS: Obstructive sleep apnoea (OSA) is associated with increased cardiovascular morbidity and mortality. Although it was previously assumed that this was due to its relation with obesity, recent data suggest that OSA is independently associated with the cardiovascular risk factors that comprise metabolic syndrome, including hypertension, insulin resistance, impaired glucose tolerance, and dyslipidaemia. However, as previous studies have only considered these variables individually, it has not been possible to determine the overall association of OSA with this syndrome. METHODS AND RESULTS: We recruited 61 male subjects with OSA and 43 controls. Glucose, insulin, lipids, and blood pressure (BP) were measured following an overnight fast. Insulin resistance was estimated using homeostasis model assessment (HOMA). Metabolic syndrome was diagnosed according to National Cholesterol Education Program (NCEP) criteria. Subjects with OSA were more obese, had higher BP and fasting insulin, were more insulin resistant, had lower HDL cholesterol, and an increased incidence of metabolic syndrome (87% vs. 35%, p<0.0001). In order to determine whether these associations were independent of obesity and other known covariates, a regression analysis adjusted for age, BMI, smoking, and alcohol consumption was performed. This demonstrated that OSA was independently associated with increased systolic and diastolic blood pressure, higher fasting insulin and triglyceride concentrations, decreased HDL cholesterol, increased cholesterol:HDL ratio, and a trend towards higher HOMA values. Metabolic syndrome was 9.1 (95% confidence interval 2.6, 31.2: p<0.0001) times more likely to be present in subjects with OSA. CONCLUSIONS: OSA is independently associated with an increase in the cardiovascular risk factors that comprise the metabolic syndrome and its overall prevalence. This may help explain the increased cardiovascular morbidity and mortality associated with this condition. PMID- 15120884 TI - Statin therapy improves cardiovascular outcome of patients with peripheral artery disease. AB - AIMS: We sought to examine the interrelationship between statin use, inflammation, and outcome of high-risk patients with advanced atherosclerosis. METHODS AND RESULTS: We prospectively studied 515 patients with severe peripheral artery disease (median age 70 years, 296 males). The cardiovascular risk profile and laboratory parameters of inflammation (high-sensitivity C-reactive protein [hs-CRP], serum amyloid A [SAA], fibrinogen, serum albumin, neutrophil counts) were obtained, and patients were followed for a median of 21 months (interquartile range 12-25) for the occurrence of myocardial infarction (MI) and death. We observed 19 MIs (5 fatal and 14 nonfatal) and 65 deaths. Cumulative survival and event-free survival rates (freedom from death and MI) at 6, 12, and 24 months were 97%, 95%, and 89%, and 96%, 93% and 87%, respectively. Patients receiving statin therapy (n=269, 52%) had a lower level of inflammation (hs-CRP p<0.001, SAA p=0.001, fibrinogen p=0.007, albumin p<0.001, neutrophils p=0.049) and better survival (adjusted hazard ratio [HR] 0.52, p=0.022) and event-free survival rates (adjusted HR 0.48, p=0.004) than patients not treated with statins. However, patients with low inflammatory activity (hs-CRP < or =0.42 mg/dl) had no significant benefit from statin therapy (p=0.74 for survival; p=0.83 for event-free survival), whereas in patients with high hs-CRP (>0.42 mg/dl) statin therapy was associated with a significantly reduced risk for mortality (adjusted HR 0.58, p=0.046) and the composite of myocardial infarction and death (adjusted HR 0.46, p=0.016). CONCLUSION: Statin therapy is associated with a substantially improved intermediate-term survival of patients with severe peripheral artery disease and a high inflammatory activity, whereas in patients with low hs-CRP no survival benefit was observed. PMID- 15120885 TI - Cellular repopulation of myocardial infarction in patients with sex-mismatched heart transplantation. AB - AIMS: Recent studies have suggested that human extracardiac progenitor cells are capable of differentiating into cardiomyocytes. In animal studies, myocardial infarction attracted bone marrow stem cells and enhanced their differentiation into cardiomyocytes. Based on these findings, we hypothesised that myocardial infarction stimulates the invasion of progenitor cells and their differentiation into endothelial and cardiac cells in the human heart. METHODS AND RESULTS: We compared autopsy samples from male control patients who had received a female donor heart with samples from such patients who developed myocardial infarction after transplantation. Fluorescence in situ hybridisation (FISH) for detection of the Y-chromosome was combined with immunofluorescence staining for CD45 and CD68 to distinguish host-derived inflammatory cells. Additionally, we used a 3D confocal imaging technique to indisputably assign Y-chromosome-positive nuclei to their cytoplasm. In patients with myocardial infarction after heart transplantation (n=5), host-derived non-inflammatory progenitor and endothelial cells were significantly increased compared to non-infarcted patients (n=9). Yet, by using this novel multi-step approach, only 0.02% of all cells were estimated to be male cardiomyocytes and their increase in infarcted regions to 0.07% was not significant. CONCLUSION: Myocardial infarction enhances the invasion of extracardiac progenitor cells and their regeneration of endothelial cells. However, a significant differentiation into cardiomyocytes as a physiological mechanism of postischaemic regeneration does not occur in transplanted patients. PMID- 15120886 TI - Coronary angiography transiently increases plasma pro-B-type natriuretic peptide. AB - AIMS: Increased plasma concentrations of B-type natriuretic peptide (BNP) and its precursor (proBNP) provide important prognostic information in patients presenting with acute coronary syndromes. Although a majority of these patients undergo early invasive assessment, the effects of coronary angiography per se on plasma BNP and proBNP concentrations are not known. We therefore sought to determine whether coronary angiography and ventriculography affect the cardiac secretion of these prognostic markers. METHODS AND RESULTS: Blood samples were collected before and two minutes after coronary angiography and ventriculography in patients with or without coronary artery disease (CAD) and normal left ventricular ejection fraction. In patients with suspected CAD and normal left ventricular ejection fraction, the plasma proBNP concentration transiently increased from 11 pmol/l (range 1-67 pmol/l) to 19 pmol/l (range 5-102 pmol/l, n=29,P<0.0001) two minutes after coronary angiography and ventriculography. The increase was similar in patients with or without CAD, although patients with stable CAD displayed higher plasma BNP and proBNP concentrations at baseline. In contrast, plasma BNP concentrations did not change after coronary angiography and ventriculography. CONCLUSION: Coronary angiography induces a transient increase in cardiac proBNP secretion. Blood sampling for plasma proBNP measurements in patient stratification and prognosis estimation should consequently be avoided immediately after coronary angiography. PMID- 15120887 TI - Beating heart against cardioplegic arrest studies (BHACAS 1 and 2): quality of life at mid-term follow-up in two randomised controlled trials. AB - AIMS: Off-pump coronary bypass grafting (OPCAB) has short-term benefits compared to conventional bypass grafting using the heart-lung machine (CABG-CPB) but may compromise longer term outcome. We aimed to compare generic and disease specific quality of life (QoL) two to four years after surgery in participants in two randomised controlled trials of OPCAB vs. CABG-CPB. METHODS AND RESULTS: Trial participants were sent four questionnaires (SF-36, EuroQol/EQ5D, Seattle Angina Questionnaire (SAQ) and Coronary Revascularisation Outcome Questionnaire (CROQ)) to assess generic and disease-specific quality of life (QoL). Of 401 participants, 22 (5.5%) had died; of the 379 survivors, 328 responded (86.5%; 159 CABG-CPB and 169 OPCAB). Median duration of follow-up was three years. QoL scores for both groups were very similar and differences between groups were not significant (p>0.05 for all questionnaires and dimensions). Summary SF-36 scores showed poorer than normal physical QoL but normal mental QoL. Among all responders, there was a tendency for CROQ scores (core total, physical and psychosocial functioning and satisfaction with treatment) to deteriorate with time after the operation (p< or =0.05). CONCLUSION: Two to four years after surgery, patients randomised to OPCAB and CABG-CPB had similar symptoms, generic and disease-specific QoL. PMID- 15120888 TI - Effects of pravastatin on coronary events in 2073 patients with low levels of both low-density lipoprotein cholesterol and high-density lipoprotein cholesterol: results from the LIPID study. AB - AIMS: Fibrates or nicotinic acid are usually recommended for secondary prevention of coronary heart disease in patients with low plasma levels of both low-density lipoprotein cholesterol (LDL-C) < or =140 mg/dL (< or =3.6 mmol/L) and high density lipoprotein cholesterol (HDL-C) < or =40 mg/dL (< or =1.03 mmol/L). The LIPID trial, a randomised, placebo-controlled trial in 9014 patients at 87 centres in Australia and New Zealand, provided an opportunity to investigate the effects of an HMG-CoA reductase inhibitor in patients with low LDL-C and low HDL C. METHODS AND RESULTS: Participants in this post hoc substudy were 2073 patients aged 31-75 years with baseline LDL-C < or =140 mg/dL (< or =3.6 mmol/L), HDL-C < or =40 mg/dL (< or =1.03 mmol/L), and triglyceride < or =300 mg/dL (< or =3.4 mmol/L). The relative risk reduction with pravastatin treatment was 27% for major coronary events (95% CI 8-42%), 27% for coronary heart disease mortality (95% CI 0-47%), 21% for all-cause mortality (95% CI 0-38%), and 51% for stroke (95% CI 24 69%). The number needed to treat to prevent a major coronary event over 6 years was 22. CONCLUSIONS: Treatment with pravastatin in patients with both low LDL-C and low HDL-C significantly reduced major coronary events, stroke, and all-cause mortality. The level of HDL-C is crucial to the risk of recurrent CHD events and, consequently, the benefit of lowering LDL-C. PMID- 15120890 TI - Value and limitations of aortic valve resistance with particular consideration of low flow-low gradient aortic stenosis: an in vitro study. AB - BACKGROUND: The calculation of valve resistance (R) rather than aortic valve area (AVA) has been proposed for the assessment of aortic stenosis (AS), based on the claim that it is less flow-dependent. Even more importantly, valve resistance has been reported to distinguish between truly severe and "pseudosevere" AS in patients with low cardiac output. However, the diagnostic value of valve resistance remains controversial. METHODS AND RESULTS: Models of stenotic aortic valves (plates and nozzles) and biological stenotic valves were studied in a pulsatile in vitro circuit using Doppler ultrasound and direct pressure and flow measurements. Anatomic AVAs ranged from 0.5 to 1.25 cm2; cardiac output varied from 1.8 to 9.0 l/min. Effective AVA was calculated with the continuity equation. The orifices of the biological valves were recorded with a video camera for planimetry. In low flow-low gradient AS, truly severe stenosis was defined by an AVA remaining <0.85 cm2 after flow normalisation, whereas AVA increased beyond 0.85 cm2 in pseudosevere AS. In rigid stenoses, valve resistance increased significantly with flow, while in bioprostheses this flow dependence was partially masked by an actual increase of the anatomic orifice area. In low flow low gradient AS, valve resistance was significantly smaller in pseudosevere AS compared to truly severe AS (129 +/- 28 vs. 176 +/- 33 dyne s cm(-5); p<0.001) at a similar baseline effective AVA. After the exclusion of datasets with mean gradients <15 and >35 mmHg, the difference in valve resistance between truly severe and pseudosevere AS was no longer significant (162 +/- 26 vs. 141 +/- 22 dyne s cm(-5); p=0.08). Nevertheless, valve resistance <120 dyne s cm(-5) was found only in pseudosevere stenoses while valve resistance >180 dyne s cm(-5) marked truly severe stenosis. CONCLUSIONS: Valve resistance is flow-dependent and not superior to calculated AVA for the assessment of AS. In low flow-low gradient AS, valve resistance <120 dyne s cm(-5) identifies pseudosevere AS, whereas valve resistance >180 dyne s cm(-5) implies truly severe AS. However, values between 120 and 180 dyne s cm(-5) are nondiagnostic, requiring repeated AVA calculations after flow normalisation. PMID- 15120889 TI - Breastfeeding and cardiovascular mortality: the Boyd Orr cohort and a systematic review with meta-analysis. AB - AIMS: To investigate the association of breastfeeding with all-cause, cardiovascular, and ischaemic heart disease mortality. METHODS AND RESULTS: A long-term follow-up of 4999 children originally surveyed from 1937 to 1939 was undertaken (Boyd Orr cohort). Four thousand three hundred and seventy-nine subjects (88%) were traced in adulthood and 3555 (71%) had complete data on all covariates. The results were combined with a meta-analysis of the published literature. In the Boyd Orr study, there was little evidence that breastfeeding was associated with all-cause (hazard ratio: 1.04 [95% CI: 0.90-1.20]), cardiovascular (1.04 [0.83-1.30]), or ischaemic heart disease (1.02 [0.77-1.36]) mortality, compared with bottle-feeding. Meta-analyses of observational studies showed little evidence of an association of breastfeeding with all-cause (pooled rate ratio: 1.01 [95% CI: 0.91-1.13]) or cardiovascular (1.06 [0.94-1.20]) mortality. There was a moderate-to-high degree of between-study heterogeneity for the association between breastfeeding and ischaemic heart disease mortality (I2 value-indicating the degree of between-study variation attributable to heterogeneity-66%), and estimates were consistent with both an important beneficial or adverse effect of breastfeeding. CONCLUSION: There is little consistent evidence that breastfeeding influences subsequent all-cause or cardiovascular disease mortality. Results from other well-designed cohorts may clarify residual uncertainty. PMID- 15120892 TI - Major changes in the statutes of the European Society of Cardiology. PMID- 15120891 TI - The sequential changes in myocardial thickness and thickening which occur during acute transmural infarction, infarct reperfusion and the resultant expression of reperfusion injury. AB - AIM: Successful primary PTCA (with TIMI 3 reflow) in patients with acute transmural infarction has been observed to result in an immediate abnormal increase in wall thickness associated with persisting abnormal post-systolic thickening. To understand the sequential changes in regional deformation during: (i) the development of acute transmural infarction, (ii) upon TIMI grade 3 infarct reperfusion and (iii) during the subsequent expression of reperfusion injury the following correlative experimental study was performed in a pure animal model in which there was no distal dispersion of thrombotic material causing either no reflow or secondary microvascular obstruction. METHODS: In 10 closed-chest pigs, a 90 min PTCA circumflex occlusion was used to induce a transmural infarction. This was followed by 60 min of TIMI 3 infarct reperfusion. M-mode ultrasound data from the "at risk" posterior wall infarct segment and from a control remote non-ischemic septal segment were acquired at standardized time intervals. Changes in regional deformation (end-diastolic (EDWT), end-systolic (ESWT) and post-systolic (PSWT) wall thickness, end-systolic strain (epsilonES) and post-systolic strain (epsilonps)) were measured. RESULTS: In this pure animal model of acute transmural infarction/infarct reperfusion (with no pre-existing intra-luminal thrombus), the induced changes in wall thickness and thickening were complex. During prolonged occlusion, after an initial acute fall in ESWT, there was no further change in systolic deformation to indicate the progression of ischaemia to infarction. Both transmurally infarcted and reperfused-infarcted myocardium retained post-systolic thickening indicating that this parameter, taken in isolation, is not a consistent marker of segmental viability and, in this regard, should be interpreted only in combination with other indices of segmental function. The most striking abnormality induced by reperfusion was an immediate increase in EDWT which then increased logarithmically over a 60 min period as reperfusion injury was further expressed. PS did not change significantly during reperfusion. Histology confirmed the wall thickness changes on reperfusion to be due to massive extra-cellular oedema. CONCLUSIONS: The identification of an acute increase in regional wall thickness in a reperfused infarct zone by cardiac ultrasound following primary PTCA might be used in patients to both identify successful infarct reperfusion and to monitor the presence, extent and resolution of the oedema associated with reperfusion injury. PMID- 15120893 TI - Studies on controlled release effervescent osmotic pump tablets from Traditional Chinese Medicine Compound Recipe. AB - A controlled release effervescent osmotic pump tablet (EOPT) of Traditional Chinese Medicine Compound Recipe (TCMCR), named Fuzilizhong prescription which includes acidic drugs consisted of many known and unknown effective components and has been used for several thousands years, was successfully prepared with sodium chloride, sodium hydrogen carbonate and hydroxypropylmethylcellulose(HPMC) as osmotic agents. Since the osmotic pressure in EOPT with sodium chloride and sodium hydrogen carbonate increased greatly, which was induced mostly by gas carbon dioxide generating from the reaction of sodium hydrogen carbonate and the acidic drugs in TCMCR after the fluid being imbibed into the compartment through the semipermeable membrane and the in vitro accumulative dissolution percent from prescription 3 was up to 96.6% at 14 hour, the problem that water insoluble drugs can not to be elementary osmotic pump tablet for its low dissolution rate was solved in the paper. On the basis of prescription 3, the drug in effervescent osmotic pump tablet was released controllably after HPMC was selected as retarder and has a good in-vitro-in-vivo correlation(IVIVC, r=0.9550). Threrfore, it could be concluded that the formulation of TCMCR is appropriate to being made into EOPT, which improves acidic drugs composed of soluble and poorly soluble components release more greatly and controllably. From the point of this, water insoluble drugs can be designed to elementary osmotic pump tablet for more complete dissolution release. PMID- 15120894 TI - Indomethacin release from ion-exchange microspheres: impregnation with alginate reduces release rate. AB - Ion-exchange microspheres (MS) designed as a drug delivery system for embolization coupling ability to occlude vessels and chemotherapy were used to evaluate a manufacturing process allowing to control the drug release rate through reduction of diffusion rate of the drug within the particle by impregnation of calcium alginate inside the porous MS. Impregnation was performed by diffusion of sodium alginate inside DEAE-Trisacryl(R) MS, dispersion of the MS in deionised water and gelling alginate by adding CaCl(2) to the dispersed MS. Studied parameters were alginate concentration, alginate diffusion time and calcium concentration. Indomethacin was loaded into the MS by eluting an aqueous indomethacin solution through a chromatographic column packed with impregnated MS. Indomethacin loading was reduced by alginate. Swelling studies showed indomethacin loading enhanced the hydrophobicity of MS while impregnation had no effect. This had an incidence on indomethacin release rate, which was assessed using the rapid elution of PBS through loaded impregnated MS packed in a column. Indomethacin loading reduced its own rate of release. MS impregnated with 2% w/v alginate gelled with a 40 mM calcium solution presented the lower release rate. This work indicated the manufacturing conditions to display a calcium alginate matrix effect on indomethacin release from DEAE-Trisacryl MS. PMID- 15120895 TI - pDMAEMA is internalised by endocytosis but does not physically disrupt endosomes. AB - Earlier workers proposed that poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) facilitates cell transfection by being endocytosed, complexed with DNA, and subsequently acting as a "proton sponge" to burst endosomes/lysosomes and release DNA to the cytosol. It also seemed feasible that the cytotoxicity of pDMAEMA might result from lysosomal bursting, which can induce cell death. Experiments were performed to determine the extent of cytotoxicity of uncomplexed pDMAEMA, the mode of cell death it induces (i.e. apoptosis or necrosis), its mechanism of entry into cells, and its ability to disrupt endosomes/lysosomes and release molecules into the cell cytosol. The results indicate that (i). pDMAEMA is highly cytotoxic and induces rapid, primarily necrotic cell death, (ii). it is internalised into cells via fluid-phase endocytosis, and (iii). although pDMAEMA affected the morphology of late endosomes/lysosomes, it did not physically disrupt them to release their contents to the cytosol. The lack of endosomal disruptive activity suggests that this is not involved in the cytotoxicity of pDMAEMA or in its ability to transfect cells. Further work will be required to establish the molecular mechanism(s) by which pDMAEMA facilitates transfection. PMID- 15120896 TI - Investigation on the release of fluorescent markers from w/o/w emulsions by fluorescence-activated cell sorter. AB - The mechanism of release of two fluorescent markers, fluorescein isothiocyanate bovine serum albumin (FITC-BSA) and fluorescein, from water-in-oil-in-water (w/o/w) emulsions was investigated using a rapid and sensitive method based on fluorescence-activated cell sorting (FACS). The release of FITC-BSA from a w/o/w emulsion was controlled by diffusion rather than by simple breakdown of the multiple droplets or by formation of reverse micelles in the oil phase. In contrast, the release of fluorescein from a double emulsion was controlled by formation of reverse micelles rather than by diffusion or simple breakdown of multiple droplets. A significant difference in the yield and fraction of FITC-BSA and fluorescein released from double emulsions was observed due to their different molecular structure and properties. The yield of FITC-BSA incorporation in a double emulsion increased with increasing FITC-BSA concentration in the internal water phase, while the yield of fluorescein decreased with increasing concentration. The fraction of FITC-BSA released from a w/o/w emulsion after 24 h decreased with an increasing concentration of FITC-BSA in the internal phase. The w/o/w emulsion with internalized FITC-BSA was more stable than that with fluorescein, indicating its further application for sorting or enriching size controlled double droplets that contained genes and water-soluble drugs. PMID- 15120897 TI - Cyclodextrins as a potential carrier in drug nebulization. AB - The inhalation route is widely studied for many drug applications focusing on either local or systemic distributions. One matter of concern is the solubilization of hydrophobic drugs. We have studied the feasibility of using different cyclodextrins (CDs) to elaborate pharmaceutical formulations for the inhalation route and tested the short-term toxicity of such formulations administered by inhalation to C57BL/6 mice. We have shown that HP-beta-CD, gamma CD, as well as RAMEB aqueous solutions can undergo aerosolization and that the resulting droplet-size ranges are compatible with pulmonary deposition. In vivo, we have demonstrated that short-term exposure to inhaled HP-beta-CD, gamma-CD and RAMEB solutions are non-toxic after assessing bronchoalveolar lavage (BAL), lung and kidney histology, bronchial responsiveness to methacholine and blood urea. The only change noted is a slight increase in lymphocyte count in the BAL after HP-beta-CD and gamma-CD inhalation. We conclude that CDs are useful in significantly enhancing the solubility of apolar drugs with a view to inhalation therapy although an increase in lymphocyte counts in the BAL after CDs inhalations needs further investigations. PMID- 15120898 TI - Poly(ether-anhydride) dry powder aerosols for sustained drug delivery in the lungs. AB - A new family of biodegradable ether-anhydride polymers was used to develop microparticles capable of controlled drug release and inhalation as a dry powder. The polymers are composed of various ratios of sebacic acid (SA) (to render the polymer insoluble in water) and poly(ethylene glycol) (PEG) (to reduce particle clearance by macrophages and improve aerosolization). Particle aerodynamic diameter was controlled within the respirable range by producing geometrically large, but low density particles as a first step toward reducing particle adhesion forces that limit efficient aerosolization of dry powders. Particles made from a variety of polymer compositions possessed high emitted doses (>80%) from a Spinhaler dry powder inhaler (DPI). Control over particle surface and bulk properties (surface roughness, surface charge, density and water retention) was achieved by varying the percentage of PEG in the polymer backbone. The addition of 10% PEG into the polymer backbone significantly enhanced deposition in the lower stages of an in vitro lung model following aerosolization from the DPI (fine particle fractions [FPF] reached 30%). Efficient aerosolization from an obsolete DPI combined with the ability to evade phagocytic clearance and provide controlled release of various drug molecules make these particles promising for prolonged drug delivery in the lung. PMID- 15120899 TI - Lipid-drug conjugate nanoparticles of the hydrophilic drug diminazene cytotoxicity testing and mouse serum adsorption. AB - Sleeping sickness is a widely distributed disease in great parts of Africa. It is caused by Trypanosoma brucei gambiense and rhodiense, transmitted by the Tse-Tse fly. After a hemolymphatic stage, the parasites enter the central nervous system where they cannot be reached by hydrophilic drugs. To potentially deliver the hydrophilic antitrypanosomal drug diminazene diaceturate to the brain of infected mice, the drug was formulated as lipid-drug conjugate (LDC) nanoparticles (NP) by combination with stearic- (SA) and oleic acid (OA). To estimate the in vivo compatibility, the particles were incubated with human granulocytes. Because as potential delivery mechanism the absorption of specific serum proteins (ApoE, Apo AI and Apo AIV) was found to be responsible for the delivery of nanoparticles to the brain, demonstrated using PBCA nanoparticles coated with polysorbate 80 (LDL uptake mechanism) the nanoparticles were incubated with mouse serum and the adsorption pattern was determined using the 2-D PAGE technique. As a result of this study, the cytotoxic potential was shown to decrease when diminazene is part of the particle matrix compared to pure fatty acid nanoparticles and the mouse serum protein adsorption pattern differs from the samples studied earlier in human serum. Especially, the fact concerning Apo-E that could be detected when the particles were incubated in human serum is absent after the mouse serum incubation, potentially, is a critical point for the delivery via the LDL-uptake mechanism but the data demonstrate that LDC nanoparticles, with 33% (wt/wt) drug loading capacity possess the potential to act as a delivery system for hydrophilic drugs like diminazene diaceturate and that further studies have to demonstrate the usability as a brain delivery system. PMID- 15120900 TI - Importance of single or blended polymer types for controlled in vitro release and plasma levels of a somatostatin analogue entrapped in PLA/PLGA microspheres. AB - The aim of the work was to develop biodegradable microspheres for controlled delivery of the somatostatin analogue vapreotide and maintenance of sustained plasma levels over 2-4 weeks after a single injection in rats. Vapreotide was microencapsulated into end-group capped and uncapped low molecular weight poly(lactide) (PLA) and poly(lactide-co-glycolide) (PLGA) by spray-drying and coacervation. Microspheres were prepared from single and blended (1:1) polymer types. The microparticles were characterized for peptide loading, in vitro release and pharmocokinetics in rats. Spray-drying and coacervation produced microspheres in the size range of 1-15 and 10-70 microm, respectively, and with encapsulation efficiencies varying between 46% and 87%. In vitro release of vapreotide followed a regular pattern and lasted more than 4 weeks, time at which 40-80% of the total dose were released. Microspheres made of 14-kDa end-group uncapped PLGA50:50 or 1:1 blends of this polymer with 35 kDa end-group uncapped PLGA50:50 gave the best release profiles and yielded the most sustained plasma levels above a pre-defined 1 ng/ml over approximately 14 days. In vitro/in vivo correlation analyses showed for several microsphere formulations a linear correlation between the mean residence time in vivo and the mean dissolution time (r=0.958) and also between the amount released between 6 h and 14 days and the AUC(6h-14d) (r=0.932). For several other parameters or time periods, no in vitro/in vivo correlation was found. This study demonstrates that controlled release of the vapreotide is possible in vivo for a duration of a least 2 weeks when administered i.m. to rats. These results constitute a step forward towards a twice-a-month or once-a-month microsphere-formulation for the treatment of acromegaly and neuroendocrine tumors. PMID- 15120901 TI - An evaluation of transmembrane ion gradient-mediated encapsulation of topotecan within liposomes. AB - Topotecan can be encapsulated in liposomes, however little is known about the role encapsulated counter ions play in drug loading efficiency and drug release. Using 1,2-distearoyl-sn-glycero-3 phosphatidylcholine and cholesterol liposomes (55:45 mole ratio), encapsulation was achieved using manganese ion gradients (MnSO(4) or MnCl(2)), with the addition of A23187, a divalent cation/proton exchanger, to maintain a pH gradient. This methodology was compared to procedures where the pH gradient was generated by use of encapsulated (NH(4))(2)SO(4) or citrate (300 mM, pH 3.5). All methods facilitated topotecan encapsulation. Liposomes prepared in the presence of the citrate and MnCl(2) (+A23187) exhibited reduced loading capacities. Liposomes prepared in the presence of (NH(4))(2)SO(4) and MnSO(4) (+A23187) could be used to generate liposomes exhibiting a drug-to lipid ratio of 0.3 (wt/wt) with an encapsulation efficiency of >90%. In vitro drug release data suggested that the (NH(4))(2)SO(4) and MnSO(4) (+A23187) formulations released drug at a reduced rate. For these formulations, the drug release rates decreased as the drug-to-lipid ratio (wt/wt) increased from 0.1 to 0.2. Cryo-electron micrographs indicated that encapsulated topotecan precipitated as linear particles within liposomes. The stability of topotecan loaded liposomes appeared to be dependent on the presence of both a pH gradient and encapsulated sulfate. PMID- 15120902 TI - Sustained delivery of vascular endothelial growth factor with alginate beads. AB - Therapeutic angiogenesis is the growth of blood vessels from a pre-existing vasculature for clinical applications such as treating myocardial and limb ischemia. Vascular endothelial growth factor (VEGF) is a potent signal transduction molecule that acts specifically on vascular endothelial cells. Encapsulation of VEGF in a polymer matrix not only protects protein against enzymatic degradation in the body, but also allows proteins to be released at a controllable rate into a localized area. In this study, VEGF was encapsulated in calcium alginate beads by the extrusion/external gelation method, and was subsequently released in PBS and in serum media. The objective was to optimize VEGF encapsulation yield and obtain VEGF release at a constant rate from alginate matrices in vitro. The incorporation of low concentrations of VEGF and NaCl can increase encapsulation yield to 97%. The rate of VEGF release from alginate beads was higher in serum than in PBS, which was due to the capacity of the serum in reducing the electrostatic interaction between alginate and VEGF. The presence of CaCl(2) in the release supernatant can shield the alginate interaction with VEGF, and a constant release rate of 6 ng/ml/day may be sustained for 14 days. These results suggest that the alginate-VEGF delivery system may be useful in the development of vascular tissue engineering and wound healing applications. PMID- 15120903 TI - Binding and detachment dynamics of microbubbles targeted to P-selectin under controlled shear flow. AB - This study was performed to assess the binding kinetics of a targeted microbubble contrast agent exposed to shear stress. An ultrasound contrast targeted to P selectin was designed by conjugating monoclonal antibodies against murine P selectin (RB40.34) to the lipid monolayer shell of the microbubble using poly(ethylene glycol)-biotin-streptavidin. The attachment and detachment of targeted microbubbles to P-selectin immobilized on a culture dish were assessed in a parallel-plate flow chamber. Targeted microbubbles (5 x 10(6) particles/ml) drawn through the flow chamber coated with P-selectin (109 sites/microm(2)) at a shear stress of 0.3 dyn/cm(2) accumulated at a rate of 565 mm(-2) min(-1). Attachment rates increased at higher plate surface densities of P-selectin, and microbubble detachment was reduced. Accumulation rate first increased with shear stress, reached a maximum at approximately 0.6 dyn/cm(2) and then decreased. Control experiments on a plate that lacked P-selectin, or was blocked with mAb RB40.34, resulted in minimal bubble attachment. Microbubble detachment was tested by ramping up shear stress at 30-s intervals. Half-maximal detachment was reached at 34 dyn/cm(2). Overall, accumulation and retention of targeted ultrasound contrast agents is possible under physiologic flow conditions and is strongly influenced by shear stress and surface density of the target receptor. PMID- 15120904 TI - Tumour cell delivery of antisense oligonuclceotides by human serum albumin nanoparticles. AB - Nanoparticles consisting of human serum albumin (HSA) and containing different antisense oligonucleotides (ASO) were prepared by desolvation. The preparation process was optimised regarding the amount of desolvating agent, stabilisation conditions as well as nanoparticle purification. The glutaraldehyde crosslinking procedure of the particle matrix was identified as a crucial parameter for biodegradability and drug release of the nanoparticles. The influence of chain length and backbone modification of ASOs on the drug loading efficiency was investigated. The loading increased with longer chain length and employment of a phosphorothioate backbone. The resulting nanoparticles were tested in cell cultures for cytotoxicity and cellular uptake. In different tumour cell lines no cytotoxic effect was observed up to nanoparticle concentrations of 5000 microg/ml. All cell lines showed a significant cellular uptake of HSA nanoparticles. The entrapment of a fluorescent labelled oligonucleotide within the particle matrix was used for the detection of the intracellular drug release of the carrier systems. Confocal laser scanning microscopy revealed that nanoparticles crosslinked with low amounts of glutaraldehyde, rapidly degraded intracellularly, leading to a significant accumulation of the ASO in cytosolic compartments of the tumour cells. PMID- 15120905 TI - Intracellular tracking of protamine/antisense oligonucleotide nanoparticles and their inhibitory effect on HIV-1 transactivation. AB - Membrane transport of antisense oligonucleotides (ODN) is an inefficient process which requires special carriers for their intracellular delivery. We have developed a delivery system for AS-ODN and their phosphorothioate analogues (AS PTO) directed against human immunodeficiency virus type 1 (HIV-1) tat mRNA for efficient transfection of HIV-1 target cells. Protamine was used to complex AS ODN and AS-PTO to form nanoparticles with diameters of about 180 nm and surface charges in the range of -18 to +30 mV. Cellular uptake of these nanoparticles was significantly enhanced compared to naked oligonucleotides. A double labeling technique with fluorescently tagged protamine and AS-ODN was used to follow the intracellular fate of the nanoparticles. Protamine/AS-ODN nanoparticles showed release of the antisense compound leading to specific inhibition of tat mediated HIV-1 transactivation. In contrast, protamine/AS-PTO complexes were stable over 72 h, and failed to release AS-PTO. These results demonstrate that protamine/AS ODN nanoparticles are useful for future therapeutical application to inhibit viral gene expression. PMID- 15120907 TI - Epigenetic regulation of the taxol resistance-associated gene TRAG-3 in human tumors. AB - TRAG-3, originally identified as a taxol resistance-associated gene from an ovarian carcinoma cell line, is upregulated in many human tumors. Like many tumor antigens, TRAG-3 mRNA is not detectable or is expressed at very low levels in normal fetal and adult human tissues except for testis, where TRAG-3 mRNA transcripts are detected abundantly. TRAG-3 mRNA is frequently overexpressed in tumors but is rarely detected in adjacent normal tissues. To delineate the transcriptional regulation of this tumor antigen, we cloned and sequenced the TRAG-3 promoter. A 539-base pair fragment upstream of the initiation site, which contains two unusual CT repeat stretches, was sufficient to drive the maximum activity of a luciferase reporter gene. Sodium bisulfite sequencing of genomic DNA revealed that the amount of DNA methylation in exon 2 and in the promoter regions is inversely correlated with gene expression. In normal tissues, TRAG-3 is hypermethylated and is thus transcriptionally silenced. In those tumors where TRAG-3 is actively transcribed, the TRAG-3 promoter and exon 2 are hypomethylated. Treatment of a TRAG-3-silenced cell line H23 with the demethylating reagent 5-aza-cytosine reduced DNA methylation and induced TRAG-3 expression in a dose-dependent manner. These results indicate that DNA demethylation is an important epigenetic mechanism that regulates the TRAG-3 tumor antigen in human tumors. PMID- 15120908 TI - Acquired Robertsonian translocations are not rare events in acute leukemia and lymphoma. AB - Robertsonian translocations are the most common constitutional structural abnormalities but are rarely reported as acquired aberrations in hematologic malignancies. The nonhomologous acrocentric rearrangements are designated as Robertsonian translocations, whereas the homologous acrocentric rearrangements are referred to as isochromosomes. Robertsonian rearrangements have the highest mutation rates of structural chromosome rearrangements based on surveys of newborns and spontaneous abortions. It would be expected that Robertsonian recombinations would be more common than suggested by the literature. A survey of the cytogenetics database from a single institution found 17 patients with acquired Robertsonian rearrangement and hematologic malignancies. This is combined with data from the literature for a total of 237 patients. All of the possible types of Robertsonian rearrangements have been reported in hematologic malignancies, with the i(13q), i(14q), and i(21q) accounting for nearly 60%. Complex karyotypic changes are seen in the majority of cases, corresponding with disease evolution. These karyotypes consistently show loss of chromosomes 5 and/or 7 in the myelocytic disorders, nonacrocentric isochromosomes, and centromeric breakage and reunion. However, nearly 25% of the acquired rearrangements were found as the sole abnormality or in addition to an established cytogenetic aberration. Most of these were the i(14q) with the myelodysplasia subtypes refractory anemia and chronic myelomonocytic leukemia. PMID- 15120909 TI - Application of bacterial artificial chromosome array-based comparative genomic hybridization and spectral karyotyping to the analysis of glioblastoma multiforme. AB - Identification of genetic losses and gains is valuable in analysis of brain tumors. Locus-by-locus analyses have revealed correlations between prognosis and response to chemotherapy and loss or gain of specific genes and loci. These approaches are labor intensive and do not provide a global view of the genetic changes within the tumor cells. Bacterial artificial chromosome (BAC) arrays, which cover the genome with an average resolution of less than 1 MbP, allow defining the sum total of these genetic changes in a single comparative genomic hybridization (CGH) experiment. These changes are directly overlaid on the human genome sequence, thus providing the extent of the amplification or deletion, reflected by a megabase position, and gene content of the abnormal region. Although this array-based CGH approach (CGHa) seems to detect the extent of the genetic changes in tumors reliably, it has not been robustly tested. We compared genetic changes in four newly derived, early-passage glioma cell lines, using spectral karyotyping (SKY) and CGHa. Chromosome changes seen in cell lines under SKY analysis were also detected with CGHa. In addition, CGHa detected cryptic genetic gains and losses and resolved the nature of subtle marker chromosomes that could not be resolved with SKY, thus providing distinct advantages over previous technologies. There was remarkable general concordance between the CGHa results comparing the cell lines to the original tumor, except that the magnitude of the changes seen in the tumor sample was generally suppressed compared with the cell lines, a consequence of normal cells contaminating the tumor sample. CGHa revealed changes in cell lines that were not present in the original tumors and vice versa, even when analyzed at the earliest passage possible, which highlights the adaptation of the cells to in vitro culture. CGHa proved to be highly accurate and efficient for identifying genetic changes in tumor cells. This approach can accurately identify subtle, novel genetic abnormalities in tumors directly linked to the human genome sequence. CGHa far surpasses the resolution and information provided by conventional metaphase CGH, without relying on in vitro culture of tumors for metaphase spreads. PMID- 15120910 TI - Genetic and pathologic significance of 1p, 17p, and 18q aneusomy and the ERBB2 gene in colorectal cancer and related normal colonic mucosa. AB - Among chromosome defects in colon cancer, deletions in 1p, 17p, and 18q have been reported as frequent events. To verify this, we investigated 1p, 17p, and 18q aneusomy in 60 colorectal cancers and their surrounding mucosa by means of fluorescence in situ hybridization (FISH). We also evaluated ERBB2 gene (alias HER-2/neu) amplification in a subset of tumors. The genetic picture in tumors was correlated with chromosomal alterations in normal colonic mucosae, as well with clinicopathologic variables. A population of cells in morphologically normal epithelium possesses genetic aberrations common to those in colon cancer, although in different percentages. No significant difference emerged in terms of fraction of nuclei with 17p monosomy between primary tumors and distal mucosal samples. Of tumor samples aneusomic for the three chromosomes, 58.3% also showed aneusomy in related normal colonic mucosa. In neoplastic samples, significant correlation existed between 1p aneusomy and mucosal component (P<0.007), between 17p aneusomy and increased depth of invasion (T3-T4) (P<0.05), and between 18q aneusomy and tumor site (P<0.03). None of the evaluated samples, neoplastic or normal, showed ERBB2 gene amplification. PMID- 15120911 TI - Combined effects of glutathione S-transferase polymorphisms and thyroid cancer risk. AB - Since exposure to ionizing radiation, a risk factor for thyroid cancer, may produce genotoxins potentially eliminated by glutathione-S-transferases, we conducted a case control study to evaluate the role of the GSTM1- and GSTT1-null genotypes and GSTP1 polymorphisms in thyroid cancer. The frequency of GSTP1 Ile/Ile, GSTM1-, and GSTT1-null genotypes was increased in cancer patients when compared with control population. Considering the genotypes over-represented in thyroid cancer patients as potential risk genotypes, we carried out an odds ratio (OR) analysis considering the presence of none, one, two, or three risk genotypes. The results obtained showed that the presence of three potentially risk alleles (GSTM1 null, GSTT1 null, and GSTP1 Ile/Ile) lead to a significant OR increase for all the cases, irrespective of the type of tumor (OR=2.91), for papillary (OR=3.64) but not for follicular tumors. The presence of GSTP1 Ile/Ile leads to a significant later age of tumor onset when compared with GSTP1 Ile/Val and Val/Val (P<0.05), suggesting a possible association between GSTP1 Ile/Ile and the age of disease manifestation. These results suggest that combined GST polymorphisms lead to a moderate increased risk for thyroid cancer, especially for the papillary type, and GSTP1 polymorphisms might modulate the age of onset of the disease. PMID- 15120912 TI - Prevalence of TEL/AML1 fusion gene in Brazilian pediatric patients with acute lymphoblastic leukemia. AB - We studied 58 childhood B-lineage acute lymphoblastic leukemia (B-ALL) in Brazilian sample patients at the time of diagnosis to investigate the prevalence of the cryptic t(12;21)(p13;q22). All bone marrow specimens were G-band karyotyped, and commercial dual-color DNA probes were used to search for fusion signals in nuclei. The karyotype analysis showed hyperdiploidy as the most frequent abnormality. The frequency of patients with TEL/AML1 gene fusion was 19% (11 out of 58 cases). Six of the positive samples had normal karyotypes. Deletion of the wild-type TEL allele was observed in 27.3% of TEL/AML1 fusion-positive cases, but it was also identified in 4.2% of the negative cases. Three cases presented two fusion signals, indicating possible duplication of the der(21). The mean age of the patients with TEL/AML1 fusion was 4.8 years and the mean amount of peripheral leukocytes was 44,270 x 10(6)/L. The higher frequency of females with B-ALL (33/58 cases) observed in our sample was probably due to the selection mode of the study cases. The prevalence of TEL/AML1 fusion in Brazilian children in our study is similar to that found in other populations. PMID- 15120913 TI - A novel t(2;20)(q35;p12) in embryonal rhabdomyosarcoma. AB - Rhabdomyosarcoma (RMS) is the most common soft-tissue sarcoma of childhood, accounting for 5%-8% of all pediatric malignancies. RMS can be categorized into several subtypes, including embryonal RMS (ERMS), the botryoid and spindle cell variants of ERMS, and alveolar RMS (ARMS). The t(2;13)(q35;q14) and the variant t(1;13)(p36;q14) are seen in a majority of ARMS cases. In contrast, the embryonal subtype of rhabdomyosarcoma has not been associated with a recurring chromosomal translocation. We describe here a novel chromosomal t(2;20)(q35;p12) occurring in a case of childhood RMS with embryonal histology. It is notable that this translocation harbors breakpoints at or near the locus of the PAX3 gene, which is involved in the most common recurring translocation associated with ARMS. PMID- 15120914 TI - A novel der(12)t(7;12)(p15;q24.3) in a patient with childhood T-cell acute lymphoblastic leukemia. AB - Approximately 35% of T-cell acute lymphoblastic leukemia (T-ALL) cases have chromosomal translocations as evaluated by conventional cytogenetic methods (G banding). Some chromosomal translocations are associated with morphologically and immunophenotypically distinct leukemia subtypes and define patients with different clinical outcomes. Chromosomal translocations may deregulate gene expression, thus contributing to the development of neoplasia, either by placing a putative oncogene under the control of strong regulatory elements or by generating chimeric genes and oncogenic fusion proteins. We report here a novel der(12)t(7;12)(p15;q24.3) in a child with T-ALL. Cloning and characterization of the breakpoint region may contribute to the discovery of new genes that are important in T-ALL. PMID- 15120915 TI - Whole genome loss of heterozygosity profiling on oral squamous cell carcinoma by high-density single nucleotide polymorphic allele (SNP) array. PMID- 15120916 TI - Identification of a novel t(1;9)(q11;q34) in acute myelocytic leukemia. PMID- 15120917 TI - Cytogenetics of a soft tissue malignant myoepithelioma. PMID- 15120918 TI - DNA copy number abnormality of oral squamous cell carcinoma detected with cDNA array-based comparative genomic hybridization. PMID- 15120919 TI - Re: Is child sexual abuse declining? Evidence from a population-based survey of men and women in Australia (Dunne, Purdie, Cook, Boyle, & Najman, 2003). PMID- 15120921 TI - Do maternal concerns at delivery predict parenting stress during infancy? AB - OBJECTIVE: In a previous study, we found that new mothers could and would express concerns about their parenting, including concerns about maltreatment and poor care. In this study, we examine the utility of early maternal concerns for predicting parenting stress in the first year. Parenting stress is important because it has been shown to be related to maltreatment and poor parent-child relationships. METHOD: A sample of 246 mothers were interviewed shortly after delivery in a publicly funded hospital about their parenting concerns, and 93% were reinterviewed in their homes about their parenting when the infants were 6 to 12 months old. Standardized measures with demonstrated psychometric properties were employed, including a measure of parenting stress due to the demands of the parenting role, characteristics of the child that make him or her difficult to care for, and stress due to difficult interactions. RESULTS: Multiple regression results indicate that both mothers concerns at delivery and sociodemographic variables are significant predictors of all three types of parenting stress in infancy. Maternal concerns were more powerful than sociodemographics in predicting stress related to the demands of parenting, while sociodemographics were more powerful for the prediction of stress related to difficult child characteristics and difficult mother-infant interaction. CONCLUSIONS: These findings suggest that knowledge of new mothers' parenting concerns might be useful for predicting parenting problems, as well as for engaging mothers' in and enhancing the effectiveness of parenting services. PMID- 15120922 TI - Childhood and adult sexual abuse, rumination on sadness, and dysphoria. AB - OBJECTIVE: The study addressed the hypothesis that adults reporting sexual abuse are more likely to exhibit a general tendency to ruminate on sadness. The relations between reported abuse, rumination on sadness, and dysphoria were also examined. METHOD: Undergraduate students (101 women and 100 men) reported on childhood and adult sexual abuse and instances of intimidation, as well as completing the Rumination on Sadness Scale [Journal of Personality Assessment 75 (2000) 404] and the Beck Depression Inventory [Depression: Clinical, Experimental, and Theoretical Aspects, Harper & Row, New York]. RESULTS: Participants who reported more abuse were more likely to report rumination on sadness. Both reports of abuse and of rumination were linked to dysphoria. Overall, causal modeling indicated that two models were equally effective in accounting for the data: (a) victimization leads to dysphoria, with this relation being partly mediated by rumination and (b) victimization leads to dysphoria, which in turn leads to rumination. Both models are consistent with prior research. For men considered separately, both models were equally effective. For women, model a best accounted for the data. CONCLUSION: One of the pathways by which victimization may lead to depression in adulthood is by encouraging the development of a tendency to ruminate on sadness. Alternatively, victimization may lead to depression by other means, and the experienced depression or dysphoria may foster rumination. PMID- 15120923 TI - A USA national survey of program services provided by child advocacy centers. AB - OBJECTIVE: Child Advocacy Centers (CACs) are designed to improve the community collaborative response to child sexual abuse and the criminal justice processing of child sexual abuse cases. CACs, in existence for 16 years, now have standards for membership developed by the National Children's Alliance (NCA) that include nine core components. And yet no systematic examination of the CAC model exists. The purpose of this paper was to assess the variations within these core components as they exist in the field. METHOD: Using a stratified random sampling design, 117 CAC directors were interviewed using a semi-structured interview that was based on the NCA's standards for membership. The eight core components of the CAC model examined in this study include: a child-friendly facility, a multidisciplinary team, an investigative child interview, a medical examination of the child, provision of mental health services, victim advocacy, case review, and case tracking. RESULTS: Results reveal the CAC model has been widely adopted by both member and nonmember centers, although variations in implementation exist. CONCLUSIONS: Future developments in the CAC model must include evaluation of the model. PMID- 15120924 TI - Interviewing youthful suspects in alleged sex crimes: a descriptive analysis. AB - OBJECTIVE: To introduce and evaluate a structured interview protocol designed for investigative interviews of youthful alleged perpetrators of child sexual abuse. METHOD: Seventy-two alleged perpetrators ranging from 9 to 14 years of age (M = 12 years) were interviewed by 1 of 13 experienced youth investigators, employed by the Israeli Ministry of Labour and Social Affairs, about incidents that had been reported by alleged victims. All interviews were conducted as part of the investigators' regular work and followed the structured interview guide appended to this article. RESULTS: Interviewers questioned older and younger children similarly, but addressed fewer invitations, directive questions, and option posing prompts to suspects who denied the allegations than to those who partially or fully admitted them. The total number of details provided by the suspects did not vary depending on their age or whether or not they fully or partially admitted the allegations. In both cases, more information was elicited using invitations rather than suggestive or option-posing prompts. CONCLUSION: Contrary to expectations, suspects who at least partially admitted their involvement provided considerable amounts of information and were very responsive to free recall prompts, although interviewers used more risky (potentially error inducing) prompts when interviewing suspects rather than alleged victims. PMID- 15120925 TI - Child sexual abuse in sub-Saharan Africa: a literature review. AB - OBJECTIVE: This article reviews the English-language literature on child sexual abuse in sub-Saharan Africa (SSA). The focus is on the sexual abuse of children in the home/community, as opposed to the commercial sexual exploitation of children. METHODS: English language, peer-reviewed papers cited in the Social Sciences Citation Index (SSCI) are examined. Reports from international and local NGOs and UN agencies are also examined. RESULTS: Few published studies on the sexual abuse of children have been conducted in the region, with the exception of South Africa. Samples are predominantly clinical or University based. A number of studies report that approximately 5% of the sample reported penetrative sexual abuse during their childhood. No national survey of the general population has been conducted. The most frequent explanations for the sexual abuse of children in SSA include rapid social change, AIDS/HIV avoidance strategies and the patriarchal nature of society. Child sexual abuse is most frequently perpetrated by family members, relatives, neighbors or others known to the child. CONCLUSIONS: There is nothing to support the widely held view that child sexual abuse is very rare in SSA-prevalence levels are comparable with studies reported from other regions. The high prevalence levels of AIDS/HIV in the region expose sexually abused children to high risks of infection. It is estimated that, approximately.6-1.8% of all children in high HIV-incidence countries in Southern Africa will experience penetrative sexual abuse by an AIDS/HIV infected perpetrator before 18 years of age. PMID- 15120926 TI - Relationship between child abuse history, trauma, and dissociation in Russian college students. AB - OBJECTIVE: The research was conducted to determine the relationship between violent trauma, child abuse history, and dissociative symptoms in a Russian population. METHOD: Three hundred and one undergraduate students from Moscow State Linguistics University participated in the study and completed the Dissociation Continuum Scale, the Violence History Questionnaire, the Traumatic Events Survey (TES), and a demographic measure. RESULTS: Scores on dissociation and its subfactors were significantly higher in the Russian sample compared to the normative US group. The best predictors for dissociation were experiencing a violent trauma, child abuse history, and/or the experience of a fearful event. Those participants with a prior child abuse history were more symptomatic after adult trauma than those with no such history. CONCLUSIONS: The relationship between trauma/abuse and dissociation is unlikely to be a result of suggestion by therapists or media exposure, since the correlation appears in a Russian population who are relatively unexposed to these suggestive sources. The validity, reliability, and structure of the dissociation measure were relatively similar in American and Russian samples. PMID- 15120927 TI - What is in a name? Refined diagnostic criteria for prognostic assessment in myelodysplastic syndrome (MDS). PMID- 15120928 TI - Philadelphia chromosome positive myelodysplastic syndrome and acute myelogenous leukemia. PMID- 15120929 TI - FLT3 mutation and response to intensive chemotherapy in young adult and elderly patients with normal karyotype. AB - The prognostic impact of FLT3 mutations on the outcome of patients with diploid AML, treated with intensive chemotherapy, was analyzed. In 176 patients, the frequency of single ITD was 30% (<61 years: 37%, >60 years: 23%), single D835 mutation 2.3%, and both 2.3%. There was no association between ITD and CR rate. ITD-positive patients <61 years had a higher frequency of resistant disease. ITD was adversely associated with CR duration and survival in both younger and elderly patients treated with comparable chemotherapy but the effect was less in the elderly. Presence of both ITD and D835 heralded the least favorable outcome. PMID- 15120930 TI - Combined stratification of refractory anemia according to both WHO and IPSS criteria has a prognostic impact and improves identification of patients who may benefit from stem cell transplantation. AB - A retrospective analysis of the relationship between the initial classification according to either FAB or WHO criteria, the presence of risk factors and the type of therapy including stem cell transplantation (SCT) on the survival was performed in a group of 106 patients with primary myelodysplastic syndrome (MDS) of FAB RA subtype. Allogeneic SCT early in the course of the disease did not significantly affect median survival in RA patients evaluated either according to FAB criteria (63.2 months in 17 SCT patients versus 64.4 months in 89 non transplanted (non-SCT) patients) or in subgroups classified separately according to WHO (64.0 months in SCT versus 91.0 months in non-SCT RA patients and 66.2 months in SCT versus 43.0 months in non-SCT refractory cytopenia with multilineage dysplasia (RCMD) patients) or International Prognostic Scoring System (IPSS) criteria despite decreased incidence of leukemic transformation (5% in SCT versus 32% in non-SCT patients). Neither univariate or multivariate analysis of different clinical and laboratory parameters revealed a significant effect of SCT on 3 or 5 years survival in RA patients. The most probable explanation was a relatively high rate of transplantation related mortality (41%) on one hand together with a slow disease progression towards leukemia (24% at 5 years in non-SCT) on the other hand. A more refined stratification of patients based on the combined WHO morphology classification and IPSS cytogenetic criteria revealed subgroup of 11 non-SCT patients with RCMD and poor karyotype with median survival significantly different from that in five SCT patients (9.2 months in non-SCT versus 89.3 months in SCT, P=0.05). Thus, combined WHO morphology/IPSS cytogenetics criteria may be helpful for identification of the high risk patients with the RA group who may benefit from early SCT despite the relatively high incidence of SCT-related complications. PMID- 15120931 TI - A phase III, randomized, double-blind, placebo-controlled, study of iseganan for the reduction of stomatitis in patients receiving stomatotoxic chemotherapy. AB - The invasion and colonization of oral cavity mucosal tissues by microflora may contribute to the pathophysiology of ulcerative oral mucositis (UOM). Iseganan is an analog of protegrin-1, a naturally occurring peptide with broad-spectrum microbicidal activity. A randomized, double-blind, placebo-controlled study was conducted to evaluate the efficacy and safety of iseganan in preventing UOM after stomatotoxic therapy. Patients received an oral rinse, consisting of iseganan 9mg or placebo, to be swished/swallowed six times daily, starting with stomatotoxic therapy and continuing up to 21 days. Patients were assessed for stomatitis and UOM, and administered a questionnaire evaluating mouth pain and difficulty swallowing thrice weekly. The primary study efficacy endpoint was the proportion of patients who did not have peak stomatitis NCI-CTC grade >or=2. Between November 2001 and June 2002, 502 patients were randomized to receive iseganan (251) or placebo (251). Equivalent numbers of patients in both cohorts received bone marrow or peripheral blood allogeneic or autologous stem cell transplantation (SCT). Forty-three percent and 37% of iseganan and placebo patients, respectively, did not have peak stomatitis grade =2 (P = 0.182). There was no significant difference between the cohorts in stomatitis severity, incidence of UOM, peak mouth pain, peak difficulty swallowing, amount of opiate analgesics used, or adverse event type or incidence. A major impact of Iseganan on reducing stomatitis, UOM, or its clinical sequelae in patients receiving stomatotoxic therapy was not detected on this study. PMID- 15120932 TI - Allelic loss during progression of follicular lymphoma. AB - We performed loss of heterozygosity (LOH) analysis on matched lymph nodes before and after progression of follicular lymphoma (FL), and found novel LOH on chromosome arm 12p. This LOH has not been previously reported in association with FL transformation. Other sites of frequent LOH include chromosome arms 6q and 9p. LOH was observed in both transformed FL and relapse FL. These data suggest that altered tumor suppressor genes exist on 6q, 9p, and 12p that have an important role in the progression of FL. Genetic changes accumulated in relapsed FL in the absence of histological changes compared to initial diagnosis. PMID- 15120933 TI - A phase II study of timed sequential therapy of acute myelogenous leukemia (AML) for patients over the age of 60: two cycle timed sequential therapy with topotecan, ara-C and mitoxantrone in adults with poor-risk AML. AB - Acute myeloid leukemia (AML) in the elderly is a serious problem characterized by poor response to therapy and short survival. To improve response to therapy, a timed sequential therapy (TST) approach was designed utilizing topotecan, cytosine arabinoside (ara-C) and mitoxantrone based on multiple studies suggesting that topotecan and mitoxantrone are effective in older patients. Thirty-two adults, >or=60-year-old (median age 69) were included. None had favorable cytogenetics and 44% had and antecedent myelodysplastic syndrome (MDS) or 2 degrees AML. Fifty-nine percent achieved a complete response (CR). Median overall survival (OS) was 6.5 months (95% confidence interval (CI): 3.1-12.0 months; range, 15 days to 25.3 months). Disease-free survival (DFS) for the 19 patients achieving a CR was 7.7 months (95% CI: 6.1-13.7 months; range, 2.9-25.3 months). There were no differences in OS or DFS between cytogenetic or disease etiology groups. Although TST was well tolerated, long-term results in this group of patients are not satisfactory and new approaches are needed. PMID- 15120934 TI - Philadelphia chromosome positive myelodysplastic syndrome and acute myeloid leukemia-retrospective study and review of literature. AB - We conducted a retrospective study to define the significance of Philadelphia chromosome (Ph) in myelodysplastic syndrome and acute leukemia in the adults at this institution and the literature was reviewed. One hundred forty-eight cases of t(9;22)(q34;q11) were identified for the period September 1993 through August 2001. The presentation of 124 cases (84%) was that of typical CML in chronic phase. Nineteen cases (13%) presented as de novo ALL, two cases (1%) presented as de novo AML and three cases (2%) presented as myelodysplastic syndrome (MDS). The estimated incidences of t(9;22)(q34;q11) in ALL and AML are 21 and 0.6%, respectively. Ph+ AMLs are increasingly being reported with either M-BCR or m-BCR gene rearrangements, similar to those found with Ph+ ALL lending support to the notion that Ph+ AMLs are distinct entities and not merely blastic phases of undiagnosed CML. This is further supported by the existence of Ph+ MDS cases. PMID- 15120935 TI - Factors influencing survival in myelodysplastic syndromes in a Brazilian population: comparison of FAB and WHO classifications. AB - The WHO classification for myelodysplastic syndromes (MDS) has introduced new categories with prognostic relevance. Our aim was to examine the predictive value of the WHO and the FAB classification compared to parameters of peripheral blood, bone marrow and IPSS. Clinical data, peripheral blood counts, bone marrow (BM) cytology and histology and survival were analyzed in consecutive newly diagnosed adult patients with MDS. All cases were diagnosed according to FAB criteria and reclassified by the WHO proposal. Among 150 patients entering the study median age was 58 years (12-90). According to FAB, 90 patients had refractory anemia (RA), 18 sideroblastic anemia, 34 refractory anemia with excess of blasts (RAEB), three RAEB-t and five chronic myelomonocytic leukemia. Using the WHO proposal, one half of the patients with RA changed category. One patient had the 5q syndrome. There were 25 cases with refractory cytopenias with multilineage dysplasia (RCMD) and 23 WHO "unclassified". These last patients presented few cell atypias, favorable IPSS and a good survival as has been described for refractory cytopenias in pediatric MDS. Hypocellular BM was found in 24% of the patients. Karyotype was available in only 85 cases. In the univariate analysis, both classifications, hemoglobin values, hypercellular bone marrow and IPSS had an influence on survival. Using the bootstrap resampling as stability test for the model created by the multivariate analysis, the WHO classification entered the model in 73%, FAB in 38% and IPSS in only 7%. Therefore, in a setting with a high number of low-risk MDS, the WHO classification is the best predictor of survival of the patients. PMID- 15120936 TI - Clinical relevance of circulating angiogenic factors in patients with non Hodgkin's lymphoma or Hodgkin's lymphoma. AB - Vascular endothelial growth factor (VEGF), basic-fibroblast growth factor (bFGF), hepatocyte growth factor (HGF), and angiogenin are important angiogenic factors. In 65 patients with non-Hodgkin's lymphoma (NHL), pre-treatment VEGF, bFGF, and HGF levels were significantly elevated compared to normal individuals, while angiogenin levels were significantly subnormal. In 37 patients with Hodgkin's disease, pre-treatment levels of VEGF and HGF were significantly elevated, bFGF levels were normal, and angiogenin levels were significantly subnormal. In patients with NHL, post-therapy levels of angiogenin were independently predictive of survival. Both pre-therapy and post-therapy VEGF levels were independently predictive of survival in patients with HD. PMID- 15120937 TI - Meta-analysis of autologous bone marrow transplantation versus chemotherapy in adult patients with acute myeloid leukemia in first remission. AB - In the present study, we have conducted a meta-analysis comparing autologous bone marrow transplantation (ABMT) and intensive chemotherapy in adult acute myeloid leukemia (AML) patients in first remission. Combined results of the six appropriate randomised controlled studies indicate that ABMT had no advantage over chemotherapy or no further treatment concerning death rate (overall rate ratio (RR)-0.95, 95% CI, 0.81-1.11), while was superior to chemotherapy concerning event rate (overall RR--0.82, 95% CI, 0.71-0.94). In conclusion, ABMT did not improve survival but it improved event-free survival (EFS) when compared with chemotherapy or no further treatment in patients with AML in first complete remission. PMID- 15120938 TI - Interleukin 11 May improve thrombocytopenia associated with imatinib mesylate therapy in chronic Myelogenous leukemia. AB - During therapy with imatinib (Gleevec), 20-30% of patients with CML in chronic phase develop grade > or =3 thrombocytopenia. This leads to treatment interruptions and dose reductions that result in a decreased probability of achieving a cytogenetic response. Interleukin-11 (oprelvekin) is a megakaryopoietic cytokine that reduces the incidence and severity of thrombocytopenia associated with chemotherapy. We report on the use of interleukin-11 in three CML patients with grade > or =3, imatinib-induced thrombocytopenia. In all three patients, interleukin-11 led to improved platelets, uninterrupted administration of imatinib and improved cytogenetic response. This observation suggests that interleukin-11 may be beneficial for patients with imatinib-induced thrombocytoepnia. PMID- 15120939 TI - Evaluation and comparison of MRP1 activity with three fluorescent dyes and three modulators in leukemic cell lines. AB - MRP1 activity was evaluated and compared in 11 cell lines with different levels of MRP1 expression using functional assays of calcein acetoxymethyl ester (calcein-AM), carboxyfluorescein diacetate (CFDA) and Rhodamine 123 (Rh123) in combination with the modulators cyclosporin A (CsA), probenecid and MK571. A good correlation was found between MRP1 expression and the modulatory effect of MK571 on calcein-AM uptake (P = 0.01 and probenecid effect on CFDA uptake (P = 0.02). Additionally, the combined modulatory effect of MK571 and probenecid on CFDA uptake (P < 0.0001) and on calcein-AM uptake (P = 0.0001) were highly significant. No correlation was found between MRP1 expression and the effects of three modulators on Rh123 uptake or efflux. In conclusion, calcein-AM and CFDA uptake assays are the best choices to probe MRP1 activity and combination of two modulators may improve the efficiency of these assays. PMID- 15120940 TI - Ectopic cyclin D1 expression blocks STI571-induced erythroid differentiation of K562 cells. AB - Bcr-Abl tyrosine kinase inhibitor induces apoptosis and erythroid differentiation of K562 cells. During this erythroid differentiation, c-Myc and cyclin D1 transcripts are transiently downregulated. Accordingly, we studied the effect of cyclin D1 overexpression on erythroid differentiation. After treatment with 250 nM STI571, 90% of K562 and 25% of K562/D1 cells underwent erythroid differentiation. The basal expression of glycophorin A in K562/D1 cells was markedly diminished compared with that by parental cells. STI571 treatment failed to induce glycophorin A expression in K562/D1 cells. During STI571 treatment, ERK activity was downregulated in parental cells, while it was constantly activated in K562/D1 cells. These results suggest that ectopic expression of cyclin D1 causes the resistance of K562 cells to erythroid differentiation by modulating ERK regulation. PMID- 15120941 TI - Platelet factor 4 enhances the adhesion of normal and leukemic hematopoietic stem/progenitor cells to endothelial cells. AB - Platelet factor 4 (PF4) is a growth regulator of hematopoietic stem/progenitor cells (HSPCs), but its role in modulating the adhesive property of normal and leukemic cells remains unclear. We used CD34(+) cord blood cells, KG1a cell line, human umbilical vein endothelial cells (HUVECs) and a transformed HUVECs ECV-304 cells to study the effect of PF4 on cell adhesion. When CD34(+) cord blood cells were cultured either in fibronectin-coated (FN) culture plate or over the layer of HUVECs for 2h, a concentration-dependent increase of the number of adhered cells was observed in the culture containing PF4. FACS analysis revealed that the treatment of PF4 resulted in an increased expression of CD49d and CXCR-4 on CD34(+) cells. Moreover, when CD34(+) cells were expanded in the presence of PF4, the adhesive ability to culture plate of CD34(+) cells was significantly increased. To elucidate the mechanism of action of PF4, KG1a cells were incubated with or without PF4 for 2h on pre-established layer of ECV-304 cells. The percentage of CD49d(+) KG1a cells increased about 1.56 +/- 0.4 fold, and that of CD54(+) ECV-304 increased about 1.7 +/- 0.6 fold. Furthermore, the mRNA expression of CD49d and CD54 was upregulated when KG1a or ECV-304 cells were incubated with PF4. The adhesion capacity of KG1a cells was reduced after incubation with the blocking monoclonal antibodies against CD49d and CD54, respectively. Our data demonstrate that PF4 is able to enhance the adhesive ability of normal and leukemia HSPCs. PMID- 15120942 TI - Severe functional alterations in vitro in CD34(+) cell subpopulations from patients with chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) arises from the malignant transformation of a hematopoietic stem cell (HSC) that gives rise to functionally defective progeny, including primitive and relatively mature progenitor cells (HPC). Both HSC and HPC are comprised within the population of CD34(+) cells, normally present in bone marrow (BM). In the present study, we have separated two different subpopulations of CD34(+) cells from CML marrow: Population I, enriched for CD34(+) Lin(-) cells; and Population II, enriched for CD34(+) CD36(-) CD38(-) CD45RA(-) Lin(-) cells, and assessed their progenitor cell content as well as their capacity to proliferate and expand in response to a combination of hematopoietic cytokines in serum- and stroma-free long-term liquid cultures. The absolute cell numbers recovered in Population I from normal and CML samples were similar; in contrast, we found that Population II from CML was amplified four fold, as compared to normal. In spite of this latter observation, no significant differences were observed in terms of the absolute number of CFC when comparing Populations I and II from CML patients and normal subjects. Interestingly, the proliferation and expansion potentials of CML cells were clearly deficient as compared to their normal counterparts. Indeed, in cultures of Population I cells the maximum fold increase in total and progenitor cell numbers corresponded to 30 and 8%, respectively, of those observed in cultures of normal marrow-derived Population I cells. Such functional deficiencies were even more evident in Population II cells in which the maximum fold increase in total and progenitor cell numbers corresponded to 3 and 0.5%, respectively, of the levels found in cultures of Population II cells from normal marrow. The present study demonstrates that bone marrow-derived CD34(+) cells from CML patients possess functional abnormalities, clearly evident in the in vitro system used by us. Among the two CML subpopulations studied here, the more immature one (Population II; enriched for CD34(+) CD36(-) CD38(-) CD45RA(-) Lin(-) cells) was the one that showed the most severe abnormalities, as compared to its relatively more mature counterpart (Population I; enriched for CD34(+) Lin(-) cells). PMID- 15120943 TI - Cross-trial networking in AML: a step forward rather than corner cutting. AB - In order to further activate the therapeutic progress against acute myeloid leukemia (AML), we recently proposed cross-trial networking based on general upfront randomization and a common standard arm. A representative standard arm of uniform treatment is recruited by random allocation of 10% patients from each participating trial. The trial own control arms are maintained and can be calibrated with the common standard arm as reference. Upfront randomization, assessment of prognostic profiles, monitoring of protocol adherence and evaluation of outcome in the standard arm are done centrally. By its design the network allows prospective comparisons and validations of specific treatment strategies across the trials, and treatments superior beyond the limits of individual trials can be identified. Recent experiences about intent-to-treat evaluation as well as a new consensus on shared protocol standards support this project. PMID- 15120944 TI - Normal bone marrow function over 6 years in a patient with dysplastic hematopoiesis and a complex karyotype. AB - Myelodysplasia associated with a complex karyotype is usually associated with advanced stage myelodysplastic syndrome (MDS) and an enhanced risk to develop secondary leukemia. We report on a 36-year-old female patient who was first presented in 1997 because of 'Pseudo Pelger-Huet' neutrophils. The remaining blood and differential counts were normal. Bone marrow examination revealed dysplasia in the erythroid and granulocytic series, no increase in blasts, and a karyotype with complex aberrations involving chromosomes 7, 13, 20 and 22. Almost all metaphases examined appeared to be affected. During the next few months, the patient was closely monitored and considered as candidate for bone marrow transplantation. However, blood counts remained stable without occurrence of significant cytopenias or an increase in blasts. Re-examinations of the bone marrow in 1998 and 1999 disclosed identical results compared to that obtained in 1997. After a total follow up of 6 years, the patient is still in good health with normal blood counts and persisting 'Pseudo Pelger-Huet' neutrophils. This exceptional case supports the notion that complex chromosomes are not invariably associated with rapid disease evolution in MDS. PMID- 15120946 TI - The epidemiology and diagnosis of bluetongue with particular reference to Corsica. AB - Bluetongue (BT) and/or BT viruses (BTV) have been identified in the Mediterranean basin and the Balkans each year from 1998 to 2002 and in particular BTV serotype 2 in the French Island of Corsica (2000 and 2001). In response to these virus incursions, the French Veterinary Authorities carried out epidemiological studies that included virological, serological and entomological analysis, and two vaccination campaigns performed in the winter of 2000/2001 and the winter and spring of 2001 and 2002. Rapid and reliable serotype differentiation is essential at the start of an outbreak to allow an early selection of vaccine to control the spread of the virus. Thus, molecular tools, that complement conventional methods, have been developed for early detection of infection, determination of the serotype, and differentiation between natural infection and vaccination. Serological results showed that the first vaccination campaign during the winter of 2000/2001 did not provide full protection for all sheep and during the summer of 2001, 335 sheep flocks in Corsica were again infected by BTV 2 (7-fold more that in 2000). Entomological studies have demonstrated that the only proven vector of the disease, Culicoides imicola, was present in the island in 2000 and that it has successfully established itself in Corsica. The safety and immunogenicity of the commercial South African vaccine were studied. Fourteen sheep were vaccinated and then observed for clinical signs. Blood, sera, spleen and lymph nodes were collected and analyzed, and the results confirmed the safety and potency of using this vaccine to protect sheep from clinical disease. As a result, an intensive vaccination campaign was performed during winter and spring 2001/2002. No cases of BT had been observed by the end of summer 2002, indicating that the vaccination campaign has been successful in protecting sheep from infection. PMID- 15120945 TI - Chronic myeloid leukemia in a patient with acquired immune deficiency syndrome: complete cytogenetic response with imatinib mesylate: report of a case and review of the literature. AB - We report a patient with acquired immune deficiency syndrome (AIDS) who developed Philadelphia-positive chronic myeloid leukemia (CML) and was successfully treated with imatinib mesylate. He achieved a complete cytogenetic response after 7 months of treatment. It appears that there is no in vivo interaction between imatinib and highly active anti-retroviral therapy (HAART) and these drugs can be concurrently administered with safety to patients with CML and AIDS. PMID- 15120947 TI - Variation in the ossification process of the anconeal and medial coronoid processes of the canine ulna. AB - This morphological and radiographic study investigates the ossification process of the anconeal and medial coronoid processes of the ulna in a sample of 142 dogs ranging in age from neonatal to 44 weeks. The anconeal process was noted to develop by appositional ossification, formation of a separate ossification center, or a combination of both. Several developmental stages of the ossification center of the anconeal process as well as its anatomic position and radiographic appearance are described. Differences have been noted in the shape of this ossification center as well as the ossification process itself. The medial coronoid process develops exclusively by appositional ossification. Unlike ossification of the anconeal process, ossification of the medial coronoid process was completed earlier (p < 0.05) in smaller than in the larger dogs. In smaller dogs, both the medial coronoid and anconeal processes were found to be mature by the age of 16 weeks. In the larger dogs, ossification of the anconeal process was completed not before 14 weeks of age and ossification of the medial coronoid process was completed about 6 weeks later. PMID- 15120948 TI - traT and CNF2 genes of Escherichia coli isolated from milk of healthy cows and sheep. AB - The objectives of the present study were to isolate Escherichia coli from milk of apparently healthy cows and sheep and to investigate the presence of traT and cytotoxic necrotising factor-2 (CNF2) virulence genes by multiplex polymerase chain reaction (PCR). Milk samples collected from a total of 1028 apparently healthy ruminants (737 cows and 291 sheep) in eastern Turkey were streaked onto 5% sheep-blood agar. E. coli was isolated and identified by biochemical tests in 5.9% (61/1028) of milk samples. Correct amplification with the molecular length of 232 bp was obtained from all E. coli isolates by the species-specific PCR. The isolation rates of the agent were calculated to be 7.6% (56/737) in cows and 1.7% (5/291) in sheep. The difference between these proportions was statistically significant (p < 0.001). Multiplex PCR showed that traT and CNF2 genes were present in 62.3% and 6.6% of all isolates, respectively. Both genes were present in 16.4% of the isolates, with only 14.7% having neither gene. PMID- 15120949 TI - The intravenous glucose tolerance test in water buffalo. AB - OBJECTIVE: The response to intravenous glucose loading in the buffalo using the intravenous glucose tolerance test (IGTT) was investigated to provide a reference for intravenous glucose injection in buffaloes. METHOD: Twelve healthy, fasted, male swamp buffaloes were divided into three groups. Group I: six buffaloes were given 50% glucose at a dosage of 1 g/kg body weight via the jugular vein. Group II: three buffaloes received normal saline. Group III: three buffaloes were not injected. Blood samples were taken from the opposite vein at 60 and 10 min pre injection (pre60 and pre10), and at 1, 5, 10, 30, 60, 120, 180, 240, 300, 360 and 420 min post-glucose injection (PGI). Plasma glucose was analyzed by the oxidase method. Insulin and glucagon were soon determined with a human radioimmunoassay kit. The insulin (pmol/l)/glucose (mmol/l) ratios (IGR) were also calculated for each sampling time. RESULTS: Mean plasma glucose, insulin and glucagon concentrations of buffaloes in groups II and III were similar at all the sampling times (p > 0.05) and the curves of the IGR for group II and group III were flat throughout. Group I Buffaloes showed an immediate 20 times increase in the mean plasma glucose concentration PGI, over the pre60 and pre10. The peak plasma insulin concentration occurred at 30 min PGI. The mean plasma glucose and insulin concentrations remained above pre-administration levels until 420 min PGI (p < 0.05). However, the mean plasma glucagon concentrations were different only at 1 and 5 min PGI sampling times. The curve of the IGR for group I showed an initial decrease at 1 min PGI, and fluctuated from 10.18 to 25.55 for the remainder of the sampling period. The correlation analysis showed that the mean plasma glucose concentration was positively correlated with insulin level (r = 0.73, p < 0.005), and significantly negatively correlated with mean plasma glucagon (r = -0.58, p < 0.05). The mean plasma insulin level did not show significant correlation with the glucagon (r = 0.06, p > 0.05). CONCLUSION: The hyperglycemia, high insulin, and protracted glucose and insulin curves, the initial decrease in the insulin/glucose ratio indicates that there was an unexpected glucose tolerance to acute intravenous glucose loading in water buffalo compared with other ruminants. The possibly suggested intravenous glucose load in buffaloes is about 5.09-8.28 mmol/l. PMID- 15120950 TI - A comparison of methods for measuring serum and urinary markers of bone metabolism in cats. AB - Biochemical markers of bone cell activity have recently been shown to be useful for monitoring skeletal health in domestic animals, including dogs and horses. The aim of this study was to evaluate a number of biochemical assays, originally developed for use in humans, for their ability to measure indicators of bone cell activity in serum and urine of normal cats over a range of ages. Bone alkaline phosphatase (BAP), a marker of bone formation, was measured in serum using wheatgerm lectin precipitation (WGL) and by ELISA. The curve derived from serial dilution of feline serum was parallel with the ELISA standard curve, indicating species cross-reactivity, and there was a significant relationship between assays (rs = 0.97, P < 0.001). Deoxypyridinoline (DPD), a marker of bone resorption, was measured in its total form in urine by HPLC and ELISA, and in its free form in serum and urine by ELISA. The dilution curve for free DPD in urine showed parallelism with the assay standard curve; however, the curves for total DPD in urine and serum did not. A significant relationship was established between total urinary DPD (HPLC) with total serum DPD (rs = 0.69, P < 0.001), and with free urinary DPD (rs = 0.95, P < 0.001) concentrations. Carboxy-terminal telopeptide of type I collagen (CTX) concentration, another marker of bone resorption, was measured in serum and urine by ELISA, and there was a significant relationship between assays (rs = 0.82, P < 0.001). CTX could not be measured reliably using an auto-analysis method. A significant relationship was established between total urinary DPD (HPLC) with serum CTX (rs = 0.59, P < 0.05), and urinary CTX (rs = 0.65, P < 0.001) concentrations. BAP (ELISA and WGL), total urinary DPD (HPLC), urinary CTX (ELISA), and serum CTX (ELISA) concentrations were significantly inversely correlated with age (rs = -0.66, -0.88, -0.61, -0.70, and -0.51, P < 0.05 respectively). Cats under two years of age had significantly higher BAP, total urinary DPD (HPLC), and urinary CTX concentrations compared to older cats. In conclusion, this study has shown that a number of commercially available assays provide reliable methods for non-invasively monitoring bone cell activity in cats and has shown that bone turnover decreases within the first two years of life, until complete skeletal maturity is attained. Future studies can now be directed at evaluating the potential clinical application of these methods. PMID- 15120951 TI - Acute phase proteins assessment for an early selection of treatments in growing calves suffering from bronchopneumonia under field conditions. AB - Blood samples were taken from calves with respiratory disease the first day of examination for determination of the serum concentration of haptoglobin, fibrinogen, alpha-2- and gamma-globulins, and albumin. A clinical examination was performed daily for the duration of the disease. The animals were retrospectively classified in two categories: those animals requiring no treatment or antibiotics alone (group A), and antibiotics associated to anti-inflammatory drugs (group B). The serum proteins were tested in order to check whether they were able to distinguish, on the first day of clinical examination, between calves requiring anti-inflammatory treatment (group B) or not (group A). About 80% of calves were properly classified in both groups by the combined use of the two serum proteins haptoglobin and fibrinogen: these two proteins, and especially haptoglobin, were useful for the identification of calves requiring an anti-inflammatory treatment. PMID- 15120952 TI - Regulating effects of porcine interleukin-6 gene and CpG motifs on immune responses to porcine trivalent vaccines in mice. AB - In order to develop novel immunoadjuvants to boost immune response of conventional vaccines, experiments were conducted to investigate the regulating effects of porcine interleukin-6 gene and CpG motifs as the molecular adjuvants on immune responses of mice that were co-inoculated with trivalent vaccines against Swine fever, the Pasteurellosis and Erysipelas suis. Synthetic oligodeoxynuleotides containing CpG motifs were ligated into pUC18, forming recombinant pUC18-CpG plasmid. Eukaryotic plasmid expressing porcine interleukin 6 (VPIL-6) were also constructed as molecular adjuvants in an attempt to enhance levels of immune responses of mice co-administered with the trivalent vaccines in this paper. The cellular and humoral immune responses of mice were systematically analysed, and the experimental results were observed that the number of white blood cells, monocytes, granuloytes and lymphocytes significantly increased, respectively, in the mice immunized with VPIL-6, compared with those of the control; the IgG content and titre of specific antibodies to the trivalent vaccine mounted remarkably in the sera from the VPIL-6 vaccinated mice; the proliferation of lymphocytes and induced IL-2 activities were significantly increased in the vaccinated groups. The above-mentioned immune responses of mice co-inoculated with pUC18-CpG plasmid were significantly stronger than those of co inoculated with pUC18 plasmid, suggesting that the immunostimulatory effect of oligodeoxynuleotides CpG is closely connected with the number of CpG motifs. These results suggest that the porcine IL-6 gene and CpG motifs could be employed as effective immunoadjuvants to elevate immunity to conventional vaccines. PMID- 15120953 TI - Microsatellite-based genotyping of the swine lymphocyte alloantigens (SLA) in miniature pigs. PMID- 15120954 TI - Oxidative stress and non-enzymatic antioxidative status in dogs with visceral Leishmaniasis. AB - Leishmaniasis is a potentially fatal chronic protozoan disease in human, canine and rodent species. The infection by Leishmania is endemic in the Mediterranean Sea region, Africa, Asia and South America. Canine visceral leishmaniasis (CanVL) is a systemic disease caused by Leishmania infantum and Leishmania chagasi from the Leishmania donovani complex group. The blood glutathione (GSH), plasma malondialdehyde (MDA), ascorbic acid (AA), beta-carotene, retinol and ceruloplasmin levels of dogs with CanVL were investigated to establish the status of the antioxidant defense mechanism in the infected animals. Dogs diagnosed as CanVL with amastigotes in lymph node smear examination and/or antibody titers > or = 128 were used as subjects, while those with no serological response against leishmaniasis were used as healthy controls. The glutathione and retinol amounts were decreased although not significantly (p > 0.05), but the MDA levels were significantly higher in dogs with VL, suggesting increased lipid peroxidation. PMID- 15120955 TI - In vitro antimicrobial activity of orbifloxacin against Staphylococcus intermedius isolates from canine skin and ear infections. AB - The objective of the study was to evaluate the in vitro activity of orbifloxacin against Staphylococcus intermedius strains isolated in France from canine skin and ear infections. The minimum inhibitory concentrations (MICs) of orbifloxacin against 240 field S. intermedius isolates (69 skin and 171 ear isolates) ranged from 0.016 to 8 mg l(-1), with MIC50 and MIC90 equal to 0.5 and 1 mg l(-1), respectively. Only one strain, a pyoderma isolate was resistant (MIC=8 mg l(-1)). Orbifloxacin was tested at different concentrations for killing rate against five isolates obtained from pyoderma cases and against a reference strain (Staphylococcus aureus ATCC 29213). Orbifloxacin expressed a concentration dependent bactericidal activity against the S. aureus reference strain, but a time-dependent bactericidal activity against S. intermedius. Orbifloxacin induced bactericidal effect against the S. intermedius strains tested with concentrations equal to or two times MIC. PMID- 15120956 TI - Lack of gender effect on the pharmacokinetics and pharmacodynamics of dexamethasone in the camel after intravenous administration. AB - The pharmacokinetics and pharmacodynamics of dexamethasone were studied in six male and six female camels after a single intravenous dose (0.05 mgkg(-1) body weight) of dexamethasone. The pharmacokinetic parameters of the two-compartment pharmacokinetic model for female and male camels, respectively (mean+/-SEM) were as follows: terminal elimination half-lives were 8.02+/-1.15 and 7.33+/-0.80 h, total body clearances were 95.5+/-16.0 and 124.5+/-11.9 ml h(-1) per kg, volumes of distribution at steady state were 0.72+/-0.08 and 0.87+/-0.14 litre kg(-1), and the volumes of the central compartment were 0.12+/-0.02 and 0.17+/-0.02 litre kg(-1). There was no significant difference in any pharmacokinetic parameter between female and male camels. Pharmacodynamic effects were evaluated by measuring endogenous plasma cortisol, circulating lymphocytes and neutrophils numbers and were analysed using indirect pharmacokinetic/pharmacodynamic models. The estimated IC50 of dexamethasone for cortisol and lymphocytes for female and male camels were 3.74+/-0.99 and 2.28+/-1.09 and 2.63+/-0.71 and 2.41+/-0.79 ng ml(-1), respectively. The EC50 for neutrophils for female and male camels were 24.5+/-5.83 and 20.2+/-3.82 ng ml(-1), respectively. There was no significant difference in any pharmacodynamic parameter between female and male camels. Dexamethasone in urine could be detected for 4-5 days by enzyme-linked immunosorbent assay and for 3-4 days by liquid chromatography/mass spectrometry after an intravenous dose of 0.05 mg kg(-1) body weight. PMID- 15120957 TI - Haemoglobin oxygen affinity and regulating factors of the blood oxygen transport in canine and feline blood. AB - Complete dynamic oxygen equilibrium curves (OEC) on dogs and cats whole blood were measured at 33, 37 and 41 degrees C. OEC were also run at three partial carbon dioxide pressures (20, 40 and 80 mmHg) as well as at five pH levels (7.2, 7.3, 7.4, 7.5 and 7.6). 2,3- diphosphoglycerate (DPG) concentrations were determined. Results were compared to those previously published in humans, using the same experimental method [Comp. Biochem. Physiol. 106 (1993) 687]. In standard conditions (pH 7.4, pCO2 40 mmHg and temperature 37 degrees C), the partial oxygen pressure at half-saturation of haemoglobin (p50) was 30.0+/-1.3 mmHg for dogs and 34.1+/-1.8 mmHg for cats. Cat's OEC was thus rightshifted compared to dog's OEC, itself rightshifted compared to human OEC. 2,3-DPG concentrations were higher in dogs than in men until they were very low in cats. Contrary to that observed in human medicine, no significant correlation was identified between standard p50 and canine 2,3-DPG values. Influence of pH, pCO2 and temperature on the OEC was saturation dependent. In dogs, Delta log p50/Delta pH was equal to -0.370, Delta log p50/Delta log pCO2 was 0.093 and Delta log p50/Delta T was 0.020. In cats, Delta log p50/Delta pH was equal to -0.405, Delta log p50/Delta log pCO2 was 0.080 and Delta log p50/Delta T was 0.016. Practically, temperature and pH variations exert a lesser influence in domestic carnivores than in humans, effect of pCO2 being similar in both. PMID- 15120958 TI - Plasma intestinal fatty acid binding protein (I-FABP) concentrations increase following intestinal ischemia in pigs. AB - Intestinal fatty acid binding protein (I-FABP) is an intracellular epithelial protein in the intestinal mucosa of many animals. I-FABP appears in the circulation following epithelial damage, and in humans, is proven to be a parameter for damage to the mucosa. In this paper, an ELISA test designed for human I-FABP analysis was used to assay pig blood samples. The test recognized I FABP cloned from pig small intestine and expressed in Escherichia coli. Furthermore, in our experimental model of (low flow) intestinal ischemia and reperfusion a significant rise in plasma I-FABP concentrations 15-30 min after clamping of the mesenteric artery was demonstrated. This is the first report that in pigs circulating I-FABP is a useful marker for (mild) intestinal injury, and could possibly be used to monitor (intestinal) health in clinical practice. PMID- 15120960 TI - Identification of a gene expression profile that discriminates indirect-acting genotoxins from direct-acting genotoxins. AB - During the safety evaluation process of new drugs and chemicals, a battery of genotoxicity tests is conducted starting with in vitro genotoxicity assays. Obtaining positive results in in vitro genotoxicity tests is not uncommon. Follow up studies to determine the biological relevance of positive genotoxicity results are costly, time consuming, and utilize animals. More efficient methods, especially for identifying a putative mode of action like an indirect mechanism of genotoxicity (where DNA molecules are not the initial primary targets), would greatly improve the risk assessment for genotoxins. To this end, we are participating in an International Life Sciences Institute (ILSI) project involving studies of gene expression changes caused by model genotoxins. The purpose of the work is to evaluate gene expression tools in general, and specifically for discriminating genotoxins that are direct-acting from indirect acting. Our lab has evaluated gene expression changes as well as micronuclei (MN) in L5178Y TK(+/-) mouse lymphoma cells treated with six compounds. Direct-acting genotoxins (where DNA is the initial primary target) that were evaluated included the DNA crosslinking agents, mitomycin C (MMC) and cisplatin (CIS), and an alkylating agent, methyl methanesulfonate (MMS). Indirect-acting genotoxins included hydroxyurea (HU), a ribonucleotide reductase inhibitor, taxol (TXL), a microtubule inhibitor, and etoposide (ETOP), a DNA topoisomerase II inhibitor. Microarray gene expression analysis was conducted using Affymetrix mouse oligonucleotide arrays on RNA samples derived from cells which were harvested immediately after the 4 h chemical treatment, and 20 h after the 4 h chemical treatment. The evaluation of these experimental results yields evidence of differentially regulated genes at both 4 and 24 h time points that appear to have discriminating power for direct versus indirect genotoxins, and therefore may serve as a fingerprint for classifying chemicals when their mechanism of action is unknown. PMID- 15120961 TI - Differentiation of DNA reactive and non-reactive genotoxic mechanisms using gene expression profile analysis. AB - Genotoxic stress triggers a variety of biological responses including the transcriptional activation of genes regulating DNA repair, cell survival and cell death. Here, we investigated whether gene expression profiles can differentiate between DNA reactive and DNA non-reactive mechanisms of genotoxicity. We analyzed gene expression profiles and micronucleus levels in L5178Y cells treated with cisplatin and sodium chloride. The assessment of cisplatin genotoxicity (up to six-fold increase in the number of micronuclei) and gene expression profile (increased expression of genotoxic stress-associated genes) was in agreement with cisplatin mode of action as a DNA adduct-forming agent. The gene expression profile analysis of cisplatin-treated cells identified a number of genes with robust up regulation of mRNA expression including genes associated with DNA damage (i.e. members of GADD45 family), early response (i.e. cFOS), and heat shock protein (i.e. HSP40 homologue). The gene expression changes correlated well with DNA damage as measured by DNA-protein crosslinks and platinum-DNA binding. To differentiate the genotoxic stress-associated expression profile of cisplatin from a general toxic stress, we have compared the gene expression profile of cisplatin-treated cells to cells treated with sodium chloride, which causes osmotic shock and cell lysis. Although the sodium chloride treatment caused a two fold induction of micronuclei, the gene expression profile at equitoxic concentrations was remarkably distinct from the profile observed with cisplatin. The profile of sodium chloride featured a complete lack of expression changes in genes associated with DNA damage and repair. In summary, the gene expression profiles clearly distinguished between DNA reactive and non-reactive genotoxic mechanisms of cisplatin and sodium chloride. Our results suggest the potential utility of gene expression profile analysis for elucidating mechanism of action of genotoxic agents. PMID- 15120962 TI - Gene expression profiles and genetic damage in benzo(a)pyrene diol epoxide exposed TK6 cells. AB - Microarray analysis is a powerful tool to identify the biological effects of drugs or chemicals on cellular gene expression. In this study, we compare the relationships between traditional measures of genetic toxicology and mutagen induced alterations in gene expression profiles. TK6 cells were incubated with 0.01, 0.1, or 1.0 microM +/-anti-benzo(a)pyrene-trans-7,8-dihydrodiol-9,10 epoxide (BPDE) for 4 h and then cultured for an additional 20 h. Aliquots of the exposed cells were removed at 4 and 24 h in order to quantify DNA adduct levels by 32P post-labeling and measure cell viability by cloning efficiency and flow cytometry. Gene expression profiles were developed by extracting total RNA from the control and exposed cells at 4 and 24 h, labeling with Cy3 or Cy5 and hybridizing to a human 350 gene array. Mutant frequencies in the Thymidine Kinase and Hypoxanthine Phosphoribosyl Transferase genes were also determined. The 10alpha-(deoxyguanosin-N(2)-yl)-7alpha,8beta,9beta-trihydroxy-7,8,9,10 tetrahydrobenzo(a)pyrene (dG-N(2)-BPDE) adduct increased as a function of dose and was the only adduct identified. A dose-related decrease in cell viability was evident at 24 h, but not at 4 h. Cell death occurred by apoptosis. At 4 h, analysis of the gene expression profiles revealed that Glutathione Peroxidase and Gadd45 were consistently upregulated (greater than 1.5-fold and significantly (P < 0.001) greater than the control in two experiments) in response to 1.0 microM BPDE exposure. Fifteen genes were consistently down-regulated (less than 0.67 fold and significantly (P < 0.001) lower than the control in two experiments) at 4 h in cultures exposed to 1.0 microM BPDE. Genes with altered expression at 4 h included genes important in the progression of the cell-cycle and those that inhibit apoptosis. At 24 h post-exposure, 16 genes, involved in cell-cycle control, detoxification, and apoptosis were consistently upregulated; 10 genes were repressed in cultures exposed to the high dose of BPDE. Real-time quantitative PCR confirmed the differential expression of selected genes. These data suggest that changes in gene expression will help to identify effects of drugs and chemicals on molecular pathways in cells, and will provide useful information about the molecular responses associated with DNA damage. Of the endpoints evaluated, DNA adduct formation was the most sensitive indicator of DNA damage. DNA adduct formation was clearly evident at low doses, but the number of genes with significantly altered expression (P < 0.001) was minimal. Alterations in gene expression were more robust at doses associated with cellular toxicity and induction of mutations. PMID- 15120963 TI - Functional genomics of UV radiation responses in human cells. AB - The gene expression responses of MCF-7, a p53 wild-type (wt) human cell line, were monitored by cDNA microarray hybridization after exposure to different wavelengths of UV irradiation. Equitoxic doses of UVA, UVB, and UVC radiation were used to reduce survival to 37%. The effects of suramin, a signal pathway inhibitor, on the gene expression responses to the three UV wavelengths were also compared in this model system. UVB radiation triggered the broadest gene expression responses, and 172 genes were found to be consistently responsive in at least two-thirds of independent UVB experiments. These UVB radiation responsive genes encode proteins with diverse cellular roles including cell cycle control, DNA repair, signaling, transcription, protein synthesis, protein degradation, and RNA metabolism. The set of UVB-responsive genes included most of the genes responding to an equitoxic dose of UVC radiation, plus additional genes that were not strongly triggered by UVC radiation. There was also some overlap with genes responding to an equitoxic dose of UVA radiation, although responses to this lower energy UV radiation were overall weaker. Signaling through growth factor receptors and other cytokine receptors was shown to have a major role in mediating UV radiation stress responses, as suramin, which inhibits such receptors, attenuated responses to UV radiation in nearly all the cases. Inhibition by suramin was greater for UVC than for UVB irradiation. This probably reflects the more prominent role in UVB damage response of signaling by reactive oxygen species, which would not be affected by suramin. Our results with suramin demonstrate the power of cDNA microarray hybridization to illuminate the global effects of a pharmacologic inhibitor on cell signaling. PMID- 15120964 TI - Comparison of basal gene expression profiles and effects of hepatocarcinogens on gene expression in cultured primary human hepatocytes and HepG2 cells. AB - Toxicogenomics is a relatively new discipline of toxicology. Microarrays and bioinformatics tools are being used successfully to understand the effects of toxicants on in vivo and in vitro model systems, and to gain a better understanding of the relevance of in vitro models commonly used in toxicological studies. In this study, cDNA filter arrays were used to determine the basal expression patterns of human cultured primary hepatocytes from different male donors; compare the gene expression profile of HepG2 to that of primary hepatocytes; and analyze the effects of three genotoxic hepatocarcinogens; aflatoxin B(1) (AFB(1)), 2-acetylaminofluorene (2AAF), and dimethylnitrosamine (DMN), as well as one non-gentoxic hepatotoxin, acetaminophen (APAP) on gene expression in both in vitro systems. Real-time PCR was used to verify differential gene expression for selected genes. Of the approximately 31,000 genes screened, 3-6% were expressed in primary hepatocytes cultured on matrigel for 16 h. Of these genes, 867 were expressed in cultured hepatocytes from all donors. HepG2 cells expressed about 98% of the genes detectable in cultured primary hepatocytes, however, 31% of the HepG2 transcriptome was unique to the cell line. A number of these genes are expressed in human liver but expression is apparently lost during culture. There was considerable variability in the response to chemical carcinogen exposure in primary hepatocytes from different donors. The transcription factors, E2F1 and ID1 mRNA were increased three-fold and six-fold (P < 0.05, P < 0.01), respectively, in AFB(1) treated primary human hepatocytes but were not altered in HepG2. ID1 expression was also increased by dimethylnitrosamine, acetylaminofluorene and acetaminophen in both primary hepatocytes and HepG2. Identification of genes that are expressed in primary hepatocytes from most donors, as well as those genes with variable expression, will aid in understanding the variability in human reactions to drugs and chemicals. This study suggests that identification of biomarkers of exposure to some chemicals may be possible in the human through microarray analysis, despite the variability in responses. PMID- 15120966 TI - Changes in expression level of genes as a function of time of day in the liver of rats. AB - Daily, rhythmic variation in various biochemical, physiological, and behavioral events is a fundamental property of biological organization. Here, we report analysis of relative levels of gene expression in the liver of 16 Fischer 344 rats as a function of time of day. Expression levels were determined for 3906 genes using high-density oligonucleotide microarrays. Of the 3906 genes, 1171 (30%) were clearly expressed while 2735 (70%) were not expressed or the expression was too low to distinguish from background levels. The maximum estimated changes observed for most genes (1029, 88%) were less than 1.5-fold. Analysis of variance and the Kruskal-Wallis tests were used to identify 67 genes whose expression was significantly altered as a function of time of day. These significantly altered genes were classified according to their functions and fall into key cellular pathways including drug metabolism, ion transport, signal transduction, DNA binding and regulation of transcription, and immune response. PMID- 15120965 TI - Applications of microarrays with toxicologically relevant genes (tox genes) for the evaluation of chemical toxicants in Sprague Dawley rats in vivo and human hepatocytes in vitro. AB - Microarrays with toxicologically relevant genes (tox genes) have been developed in our laboratory for toxicogenomics studies in rat, dog and man. The genes were chosen using published information as well as a discovery process for genes responsive to toxic treatments using transcription profiling experiments conducted with rats and dogs. In addition to published information human tox genes were derived from rat tox genes based on gene homology. Using the microarray with rat-specific tox genes, a database containing gene expression, histopathology, and clinical chemistry findings has been generated for 89 compounds. Analysis of the database indicates that treatment with toxic compounds induces specific gene expression patterns. Dose- and time-dependent response relationships in gene expression were observed for treatment with toxic compounds. Gene expression at 24h was found to correlate well with organ toxicity observed at 72 h. Mining of the database led to the selection of specific groups of genes (predictive gene sets) whose expression patterns are predictive of organ toxicity with a high degree of accuracy (approximately 90%). The data also provide insight on toxic mechanism and gene regulation pathways. For instance, carbon tetrachloride and chloroform treatments were found to decrease the expression of the cytochrome P450 isoform 3A1 gene while enhancing the expression of the multiple drug resistance gene MDR1 in liver, clearly demonstrating that the CYP3A1 and MDR1 genes were not co-regulated as postulated by some researchers. This approach, the use of gene expression as an endpoint to define organ toxicity, is extended to the definition of human drug toxicity using primary human hepatocytes as a test system. Preliminary results demonstrate that the toxic drug, troglitazone, can be clearly distinguished from the less toxic analogues, rosiglitazone and pioglitazone based on their effects on tox gene expression in human hepatocytes. Our results with both rats in vivo and human hepatocytes in vitro suggest that microarrays with toxicologically relevant genes can be used routinely for the evaluation of chemical toxicity. PMID- 15120967 TI - Profiling of hepatic gene expression in rats treated with fibric acid analogs. AB - Peroxisome proliferator-activated receptors (PPARs) are a group of nuclear receptors whose ligands include fatty acids, eicosanoids and the fibrate class of drugs. In humans, fibrates are used to treat dyslipidemias. In rodents, fibrates cause peroxisome proliferation, a change that might explain the observed hepatomegaly. In this study, rats were treated with multiple dose levels of six fibric acid analogs (including fenofibrate) for up to two weeks. Pathological analysis identified hepatocellular hypertrophy as the only sign of hepatotoxicity, and only one compound at the highest dose caused any significant increase in serum ALT or AST activity. RNA profiling revealed that the expression of 1288 genes was related to dose or length of treatment and correlated with hepatocellular hypertrophy. This gene list included expression changes that were consistent with increased mitochondrial and peroxisomal beta-oxidation, increased fatty acid transport, increased hepatic uptake of LDL-cholesterol, decreased hepatic uptake of glucose, decreased gluconeogenesis and decreased glycolysis. These changes are likely linked to many of the clinical benefits of fibrate drugs, including decreased serum triglycerides, decreased serum LDL-cholesterol and increased serum HDL-cholesterol. In light of the fact that all six compounds stimulated similar or identical changes in the expression of this set of 1288 genes, these results indicate that hepatomegaly is due to PPARalpha activation, although signaling through other receptors (e.g. PPARgamma, RXR) or through non receptor pathways cannot be excluded. PMID- 15120968 TI - Gene expression profiling reveals multiple toxicity endpoints induced by hepatotoxicants. AB - Microarray technology continues to gain increased acceptance in the drug development process, particularly at the stage of toxicology and safety assessment. In the current study, microarrays were used to investigate gene expression changes associated with hepatotoxicity, the most commonly reported clinical liability with pharmaceutical agents. Acetaminophen, methotrexate, methapyrilene, furan and phenytoin were used as benchmark compounds capable of inducing specific but different types of hepatotoxicity. The goal of the work was to define gene expression profiles capable of distinguishing the different subtypes of hepatotoxicity. Sprague-Dawley rats were orally dosed with acetaminophen (single dose, 4500 mg/kg for 6, 24 and 72 h), methotrexate (1mg/kg per day for 1, 7 and 14 days), methapyrilene (100mg/kg per day for 3 and 7 days), furan (40 mg/kg per day for 1, 3, 7 and 14 days) or phenytoin (300 mg/kg per day for 14 days). Hepatic gene expression was assessed using toxicology-specific gene arrays containing 684 target genes or expressed sequence tags (ESTs). Principal component analysis (PCA) of gene expression data was able to provide a clear distinction of each compound, suggesting that gene expression data can be used to discern different hepatotoxic agents and toxicity endpoints. Gene expression data were applied to the multiplicity-adjusted permutation test and significantly changed genes were categorized and correlated to hepatotoxic endpoints. Repression of enzymes involved in lipid oxidation (acyl-CoA dehydrogenase, medium chain, enoyl CoA hydratase, very long-chain acyl-CoA synthetase) were associated with microvesicular lipidosis. Likewise, subsets of genes associated with hepatotocellular necrosis, inflammation, hepatitis, bile duct hyperplasia and fibrosis have been identified. The current study illustrates that expression profiling can be used to: (1) distinguish different hepatotoxic endpoints; (2) predict the development of toxic endpoints; and (3) develop hypotheses regarding mechanisms of toxicity. PMID- 15120969 TI - Integration of clinical and gene expression endpoints to explore furan-mediated hepatotoxicity. AB - Molecular techniques, such as cDNA microarrays, are being used to aid in the elucidation of the mechanisms of toxicity of a variety of compounds. In this study, we evaluate the molecular effects of furan in the rat liver. Sprague Dawley rats were exposed to 4 or 40 mg/kg furan for up to 14 days. Furan induced an initial degenerative and necrotic phenotype that was followed by inflammation and fibrosis, consistent with previous observations for this compound. RNA was harvested from each lobe of the liver at several time points to observe whether lobe-specific gene expression effects occurred. Similar gene expression changes were observed in all lobes, however the magnitude of gene expression change was more pronounced in the right lobe. Finally, to help determine the correlation between gene expression changes and liver pathology, we applied traditional microarray visualization tools to the assessment of clinical chemistry and pathology parameters. PMID- 15120970 TI - Persistence of gene expression changes in stomach mucosae induced by short-term N methyl-N'-nitro-N-nitrosoguanidine treatment and their presence in stomach cancers. AB - Cancers induced by different carcinogens show distinct expression profiles. In addition to the specific alterations of tumor-related genes induced by specific carcinogens, it is possible that some initial responses induced by a carcinogen could persist for long periods and are consistently present in the cancers induced. We have analyzed the initial responses in the rat pyloric mucosae after treatment for 2 weeks with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Gene expression was monitored 1 day, 2 weeks and 4 weeks after MNNG treatment by oligonucleotide microarray analysis. Of the differentially expressed genes showing greater than three-fold difference 1 day after MNNG treatment, 143 and 26 genes were up- and down-regulated, respectively, in MNNG-induced stomach cancers. Among these genes, 25 and 6 genes were up- and down-regulated, respectively, in the histologically normal pyloric mucosae, even 4 weeks after cessation of MNNG treatment. Among the up-regulated genes, many genes involved in tissue remodeling (Spi15, Serpine1 and Fst) and cellular growth (Bdnf, Ros1 and Fgf10) were present. The six down-regulated genes included TGF-beta-inducible early growth response gene. These findings demonstrate that some expression changes induced by MNNG persist for a prolonged period and are present in cancers. Persistent expression changes are considered to be important for prediction of past carcinogen exposure, and could provide a molecular environment favorable for malignant transformation. PMID- 15120971 TI - Gene expression profile in bone marrow and hematopoietic stem cells in mice exposed to inhaled benzene. AB - Acute myeloid leukemia and chronic lymphocytic leukemia are associated with benzene exposure. In mice, benzene induces chromosomal breaks as a primary mode of genotoxicity in the bone marrow (BM). Benzene-induced DNA lesions can lead to changes in hematopoietic stem cells (HSC) that give rise to leukemic clones. To gain insight into the mechanism of benzene-induced leukemia, we investigated the DNA damage repair and response pathways in total bone marrow and bone marrow fractions enriched for HSC from male 129/SvJ mice exposed to benzene by inhalation. Mice exposed to 100 ppm benzene for 6h per day, 5 days per week for 2 week showed significant hematotoxicity and genotoxicity compared to air-exposed control mice. Benzene exposure did not alter the level of apoptosis in BM or the percentage of HSC in BM. RNA isolated from total BM cells and the enriched HSC fractions from benzene-exposed and air-exposed mice was used for microarray analysis and quantitative real-time RT-PCR. Interestingly, mRNA levels of DNA repair genes representing distinct repair pathways were largely unaffected by benzene exposure, whereas altered mRNA expression of various apoptosis, cell cycle, and growth control genes was observed in samples from benzene-exposed mice. Differences in gene expression profiles were observed between total BM and HSC. Notably, p21 mRNA was highly induced in BM but was not altered in HSC following benzene exposure. The gene expression pattern suggests that HSC isolated immediately following a 2 weeks exposure to 100 ppm benzene were not actively proliferating. Understanding the toxicogenomic profile of the specific target cell population involved in the development of benzene-associated diseases may lead to a better understanding of the mechanism of benzene-induced leukemia and may identify important interindividual and tissue susceptibility factors. PMID- 15120972 TI - The transcriptional profile of the kidney in Tsc2 heterozygous mutant Long Evans (Eker) rats compared to wild-type. AB - Hereditary renal cell carcinoma (RCC) in Eker rats results from an inherited insertional mutation in the Tsc2 tumor suppressor gene and provides a valuable experimental model to characterize the function of the Tsc2 gene product, tuberin in vivo. The Tsc2 mutation predisposes the Eker rat to develop renal tumors at an early age. The exact mechanism of Tsc2 mediated tumor suppression is not known, however, there is evidence that it is most likely mediated by changes in cell cycle regulation via the PI3K/Akt pathway. The present study was designed to identify if gene expression was different in Tsc2 heterozygous mutant rat kidney compared to wild-type and if any of those differences are associated with tumorigenesis. cDNA microarray analysis of the untreated Tsc2 (+/-) mutant Long Evans (Eker) rat was compared to the Tsc2 (+/+) wild-type Long Evans rat to search for patterns that might be indicative of the intrinsic role of Tsc2. Of 4395 genes queried, 3.2% were significantly altered in kidneys from heterozygous mutant rats, of which 110 (76%) were up-regulated and 34 (24%) were down regulated relative to the wild-type. The genes with altered expression belonged to the functional categories of cell cycle regulation, cell proliferation, cell adhesion and endocytosis. Many of these genes appear to be directly or indirectly regulated by the PI3K/Akt pathway. In addition to the PI3K/Akt pathway, other signaling pathways were also differentially expressed in Tsc2 mutant Eker rat kidneys compared to wild-type rats. The gene expression profiles of the Tsc2 heterozygous mutant and wild-type animals highlights new pathways for investigation that may be associated with the tumorigenic activity of tuberin loss and correlate with the enhanced susceptibility of the Tsc2 mutant animal's tendency to develop renal cell carcinoma. PMID- 15120973 TI - Gene expression patterns associated with infertility in humans and rodent models. AB - Modern genomic technologies such as DNA arrays provide the means to investigate molecular interactions at an unprecedented level, and arrays have been used to carry out gene expression profiling as a means of identifying candidate genes involved in molecular mechanisms underlying a variety of phenotypes. By comparing gene expression profiles from normal and abnormal human testes with those from comparable infertile mouse models, we endeavored to identify genes and gene networks critical for male fertility. We used commercially available filter-based DNA arrays to analyze testicular gene expression from eight human testis biopsies and three different infertile mouse models (atrichosis mutation, ataxia telangiectasia knockout and CREMtau knockout). Forty-seven mouse genes exhibited differential testicular gene expression (P <0.01) associated with male infertility. These included genes involved in DNA repair (Vim, Rad23A, Rad23B), glutathione metabolism (Gsr, Gstp 1, Mgst1), proteolysis (Ace, Casp1, Ctsd), spermatogenesis (Prlr, Tmsb4 and Zfp-37) and stress response (Hsp 1, Osp94). The expression of 19 human genes was different (P<0.05) between normal and abnormal samples, including those associated with apoptosis (GADD45), gonad development (SOX9), proteolysis (PSMC3, SPINK2, TIMP3, UBE213) and signal transduction (DLK1, NAP4, S100A10). Direct comparison of differentially expressed human and mouse genes identified glucose phosphate isomerase, and the highly similar human tissue inhibitor of metalloproteinase 3 (TIMP3) and mouse Timp2. Using DNA microarrays to profile gene expression in testes from infertile animal models and humans will be useful for understanding congenital infertility, and also infertility caused by environmental exposures where the same genes and molecular mechanisms are involved. PMID- 15120974 TI - Development of public toxicogenomics software for microarray data management and analysis. AB - A robust bioinformatics capability is widely acknowledged as central to realizing the promises of toxicogenomics. Successful application of toxicogenomic approaches, such as DNA microarray, inextricably relies on appropriate data management, the ability to extract knowledge from massive amounts of data and the availability of functional information for data interpretation. At the FDA's National Center for Toxicological Research (NCTR), we are developing a public microarray data management and analysis software, called ArrayTrack. ArrayTrack is Minimum Information About a Microarray Experiment (MIAME) supportive for storing both microarray data and experiment parameters associated with a toxicogenomics study. A quality control mechanism is implemented to assure the fidelity of entered expression data. ArrayTrack also provides a rich collection of functional information about genes, proteins and pathways drawn from various public biological databases for facilitating data interpretation. In addition, several data analysis and visualization tools are available with ArrayTrack, and more tools will be available in the next released version. Importantly, gene expression data, functional information and analysis methods are fully integrated so that the data analysis and interpretation process is simplified and enhanced. ArrayTrack is publicly available online and the prospective user can also request a local installation version by contacting the authors. PMID- 15120975 TI - Chinese translation and validation of the Nowicki-Strickland Locus of Control Scale for Children. AB - The objective of this study is to evaluate the psychometric properties of the Chinese version of the Nowicki-Strickland Locus of Control Scale for Children for its utility in clinical research and nursing practice. The scale was translated from the original English language into the Chinese language and the reliability and validity of the translated scale were tested. Results showed that there was high test re-test reliability, acceptable internal consistency reliability, appropriate content validity, concurrent validity and construct validity. It is concluded that the psychometric properties of the Chinese version of the Nowicki Strickland Locus of Control Scale for Children presented in this paper supports its feasibility as a research instrument to measure children's locus of control objectively and appropriately in the Chinese population. PMID- 15120976 TI - Training needs analysis. A literature review and reappraisal. AB - Training needs analysis is the initial step in a cyclical process which contributes to the overall training and educational strategy of staff in an organisation or a professional group. The cycle commences with a systematic consultation to identify the learning needs of the population considered, followed by course planning, delivery and evaluation. Although much has been written about training needs analysis in relation to post-registration nursing education, there is disagreement concerning its impact on the training cycle and its potential to influence service delivery. This stimulated the literature review presented below. Initial searches of nursing databases identified 266 works. Twenty three (8.6%) contained empirical findings relating to post registration nursing education in which assessment of training needs was presented as the major aim. Most of these accounts were concerned with the training needs of nurses in more than one organisation and were classified as macro-level training needs analysis. However, seven studies were concerned with a single, specific organisation (micro-level training needs analysis). Despite their smaller scale and more limited scope, micro-level training needs initiatives demonstrated greater methodological rigour, were more likely to consider the stakeholder perspective, to generate findings which could positively influence the rest of the training cycle and showed the greatest potential for influencing service delivery and quality of patient care. The review drew attention to the similarities between the training cycle and the audit cycle and resulted in the development of a model which could be used to evaluate the effectiveness of the process and outcomes of future training needs analysis initiatives. PMID- 15120977 TI - Nurses' self-assessment of their nursing competencies, job demands and job performance in the Taiwan hospital system. AB - This exploratory study investigated nurses' self-assessment of their own nursing competencies, job demands and job performance in Taiwan. Nurses' self-evaluation on their own job performance was conceptualized as an indicator of nursing care quality. A total of 21 competencies were clustered into three groups: basic-level patient care skills, intermediate-level patient care and fundamental management skills, and advanced-level patient care and supervision skills. Nurse subjects were randomly selected from the member roster of Kaohsiung Nurse Association; 850 nurses were invited to participate and questionnaire packets were sent to their homes. The overall response rate was 35.8%. Multiple regression analyses found that nurses' self-assessment of intermediate patient care skills, the difference between nurses' self-assessment and job demands for basic patient care skills, and nurses' overall satisfaction with their own nursing competencies were three significant predictors of overall satisfaction with nurses' own job performance. Nurses' self-assessment on basic patient care skills and advanced patient care skills contributed to nurses' levels of overall satisfaction with their own nursing competencies. These results suggest a relationship between competency and performance. These findings may serve as a guide to amend academic nursing courses and on-job training programs as appropriate to place a greater emphasis on the competencies desired for providing high quality of nursing services. PMID- 15120979 TI - Responses and experiences after radical prostatectomy: perceptions of married couples in Switzerland. AB - The purpose of this qualitative study was to explore the responses and experiences of a sample of Swiss men after radical prostatectomy and their intimate partners. Interviews were conducted with 10 couples and analyzed using constant comparative method. Getting a grip on it was the core process that was developed. Men focused on regaining control over their lives, urinary and erectile function while wives efforts focused on being there. The results revealed that current clinical practice of follow up at 3 months may not address the serious deficits in the patient's ability to "get a grip" on incontinence and other complications of surgery. PMID- 15120978 TI - Family carers for older relatives: sources of satisfaction and related factors in Finland. AB - This study focuses on sources of satisfaction among family carers for older relatives and related factors in Finland. It is part of a major international project concerned with the coping of family carers in four countries. The results reported here are based on questionnaire data collected among 290 family carers from three Finnish towns. The questionnaire included the Carer's Assessment of Satisfaction Index (CASI) developed by Nolan and Grant (1992). The data were processed using SPSS statistical software. The results suggest that carers derived most satisfaction from seeing they could help their relative feel more comfortable, making them feel needed and wanted. Among factors relating to interpersonal dynamics, carers felt that this is one way in which they can show their love for the person they cared for. The family carer's as well as the dependent person's gender, the family carer's age, care burden and general life satisfaction were associated with sources of satisfaction. The results are compared with earlier findings on family carers' sources of satisfaction in Sweden and the UK. Implications for policy and practice are considered. PMID- 15120980 TI - Changing perceptions of womanhood: living with Parkinson's Disease. AB - This study adopted a multiple case study design to develop our understanding of the experiences and adjustments made by women with Parkinson Disease in relation to womanhood. Nineteen women participated in this investigation telling their stories through a combination of individual interviews, group interviews, reflective diaries, reflective tapes and creative writing. Data were analysed using the framework of Intrapersonal, Interpersonal, Extrapersonal and Metapersonal health proposed by Boddy and Rice (Perspectives on Health and Illness, Dunsmore Press, Palmerston North, 1992). Women reported that major changes were required in their lives in each of these aspects of health. In particular women reported labile emotions, changing body images, changing lifestyles, changing relationships with partner, family and friends, increasing dependence, decreasing role fulfillment and the need for support versus increasing isolation. The findings are of particular relevance to nurses and other health professionals involved with women with Parkinson's Disease as well as social services and voluntary agencies. PMID- 15120981 TI - Nursing and the avant-garde. AB - Through an exploration of the theory of the avant-garde, this paper explores the task for nursing in the new humanities of the 21st Century. Drawing on the theory of the avant-garde in general and the work of the French philosopher Jacques Derrida in particular, it argues that nursing must always return to its basic principles, that of the human condition (humanitus). But in doing so, it must also consider its place at the table of the humanities that are undergoing profound changes in western capitalist societies both in the education and practice sectors. It is through its connection with the humanities that the notion of the avant-garde is used to introduce the concept of reconnaissance thinking again. The purpose and usefulness of this approach for nursing lies in the argument that progress or improvement, whether it be in some aspect of care or professional issue, always involves returning to something, to think of something in a new way. Thus, although nursing still has certain issues to resolve (which I attend to below), thinking through these issues with reference to the avant-garde gives ground for optimism, towards a future that is not yet determined. PMID- 15120982 TI - Becoming a role model: the breastfeeding trajectory of Hong Kong women breastfeeding longer than 6 months. AB - While a substantial proportion of breastfeeding women stop early in the postpartum period, some women are able to breastfeed for longer periods. The aim of this research was to explore the experience of breastfeeding with a subsample of Hong Kong women who have breastfed for longer than 6 months. Participants (n=17) were recruited from a larger infant-feeding study (n=360) conducted in tertiary-care hospitals in Hong Kong. In-depth qualitative interviews were conducted and content analysis was used to analyse the data. Data analysis revealed four themes that encompassed the women's experiences: (1) making the decision, (2) maintaining family harmony, (3) overcoming barriers, and (4) sustaining lactation. Antenatally, participants anticipated that breastfeeding would be very 'difficult' and described how the practice did not fit with the image of a professional woman in Hong Kong. Despite family opposition, frequently from their mother-in-law, and lack of societal acceptance, difficulties were overcome by what the Chinese people call hung-sum or determination. This study highlights unique cultural and social findings affecting breastfeeding women in Hong Kong which may be useful to health-care providers working with Chinese women locally and internationally. PMID- 15120983 TI - Identifying research priorities in nursing and midwifery service delivery and organisation: a scoping study. AB - This paper reports on a scoping study commissioned by the National Co-ordinating Centre Service Delivery and Organisation (NCCSDO) Research and Development (R&D) to identify priorities for nursing and midwifery research funding in England and Wales. The study comprised three strands (i) nationally held focus groups with service user representatives (32, plus six written submissions) (ii) semi structured telephone interviews with 64 stakeholders (nursing, midwifery, medical, social care and allied health professionals; research commissioners; policy makers; educators; managers; researchers) and (iii) literature analysis. Five priority areas for research were identified-appropriate, timely and effective interventions; individualised services; continuity of care; staff capacity and quality; user involvement and participation-from which exemplar research questions and issues for strategic commissioning are suggested. PMID- 15120984 TI - Woundcare Research for Appropriate Products (WRAP): validation of the TELER method involving users. AB - Woundcare Research for Appropriate Products (WRAP) is a novel collaboration WRAP between industry and clinicians, funded by the Engineering and Physical Research Sciences Council. WRAP objectives included the development and testing of methodologies to identify patients' and clinicians' needs with respect to wound dressings for exudate management. The management of exudate was the focus because it was demonstrated to be the pivotal problem for patients and clinicians in a study of malignant wounds, and is a recurring problem in other wound types. A clinical note-making system (Treatment Evaluation by Le Roux's method--TELER) was validated as a method of collecting observational data of dressing performance in the context of total patient care, thereby involving the users of dressing products. The validation process was a form of consensus where multiple sources of data were used to define patient problems, within the TELER indicators, to measure a change or lack of change in the problems during a period of treatment and care and to draw conclusions about dressing performance and patient experiences. PMID- 15120985 TI - The association between depressive symptoms and social support in Taiwanese women during the month. AB - The purpose of the study was to explore the association between depressive symptoms and social support in Taiwanese women doing the month. A correlational survey design using the Postpartum Social Support Questionnaire (PSSQ) and the Edinburgh Postnatal Depression Scale (EPDS) to measure social support and postnatal depressive symptomatology was employed. Two hundred and forty postpartum women receiving care in two teaching hospitals in Taipei, Taiwan, aged between 20 and 35, with no peri-natal complications or previous psychiatric history, experiencing a normal spontaneous delivery of one full term healthy baby, were selected. Each was mailed the PSSQ and the EPDS as well as a short, semi-structured self-report questionnaire requesting demographic details and subjective data relating to the experience of doing the month and depressive symptoms during the fourth week following birth. One hundred and eighty six women (78%) returned questionnaires. Taiwanese postpartum women were less depressed when they stayed in their parents' home and had their own mothers take care of them. It was found that the greater the level of postpartum social support received by the women doing the month, the lower the risk of postnatal depressive symptoms experienced. Almost a quarter (24%) of the variance of the symptoms was attributed to dissatisfaction with parents' instrumental support and unwanted emotional support from parents-in-law. It is concluded that the ritual of doing the month provides valuable social support and may help to prevent postnatal depression in Taiwanese women. PMID- 15120986 TI - The cell nucleus. PMID- 15120987 TI - "On the move"ments of nuclear components in living cells. AB - The use of GFP fusion proteins has dramatically changed our view of how the cell nucleus is organized and how functions are carried out. In this review we focus on recent advances related to the dynamics of chromatin domains, as well as the dynamics of nuclear proteins and several nuclear organelles. PMID- 15120988 TI - Nuclear export of mRNA: from the site of transcription to the cytoplasm. AB - Cellular mRNAs are produced in the nucleus and must be exported to the cytoplasm to allow for their translation into proteins. Recruitment of export factors to nascent mRNA starts cotranscriptionally and involves elaborate systems of quality control. Correctly processed mRNAs are committed for export in the form of large ribonucleoprotein complexes (mRNPs). Translocation of mRNPs through the nuclear pore complex (NPC) is mediated by a conserved heterodimeric transport receptor (NXF1/p15 in metazoa and Mex67p/Mtr2p in yeast) that bridges the interaction between the mRNP and the NPC. In this review, we describe the cis- and trans requirements for mRNA export as well as the different mechanisms of recruiting export factors to mRNPs. We also discuss the significance of linking mRNA export with both downstream and upstream events in gene expression. PMID- 15120989 TI - Assembly of kinetochores in vertebrate cells. AB - The centromere is a specialized region of each chromosome that is essential for faithful chromosome segregation during mitosis and meiosis in eukaryotic cells. Centromeres are the site at which kinetochores are formed. The kinetochore is responsible for microtubule binding and chromosome movement. In this review, I will focus on recent advances in our understanding of centromere DNAs as sites for kinetochore assembly and the mechanism underlying kinetochore assembly in vertebrate cells. PMID- 15120990 TI - Structure in the amphibian germinal vesicle. AB - The germinal vesicle (GV) of Xenopus laevis is an enormous nucleus that contains 18 giant lampbrush chromosomes and thousands of inclusions. The inclusions are primarily of three types: approximately 1500 extrachromosomal nucleoli, 50-100 Cajal bodies, and several thousand B-snurposomes, which correspond to speckles or interchromatin granule clusters in other nuclei. The large size and abundance of the GV organelles, as well as the ease with which they can be studied both in vivo and in vitro, make the GV an ideal object for analysis of nuclear structure and function. PMID- 15120991 TI - Mitotic chromosome formation and the condensin paradox. AB - During cell division, the chromatin is compacted and resolved into discrete mitotic chromosomes whose proper formation is essential for the faithful distribution of the replicated genome to the daughter cells. Chromatin within mitotic chromosomes is packaged in an orderly and reproducible fashion, but the nature of this higher-order structure has remained elusive, as have the mechanisms of its establishment. Here we provide an overview of how the functional dissection of a non-histone protein complex, condensin, has contributed to our understanding of mitotic chromosomes. Recent studies have revealed that mitotic chromosome formation involves two events: chromatin compaction and establishment of a stable intrinsic higher-order structure. Surprisingly, condensin is only required for the second of these events. PMID- 15120992 TI - Ribosome biogenesis: of knobs and RNA processing. AB - The synthesis of ribosomes in eukaryotes involves processing of pre-ribosomal RNA (pre-rRNA) and sequential assembly of a large number of ribosomal proteins on the rRNAs. Although we have gained tremendous insights into the processing of pre rRNA intermediates in the last three decades, little was known about the dynamic nature of ribosome biogenesis. Only recently the development of efficient affinity-purification procedures and mass-spectrometry techniques has allowed the isolation of large pre-ribosomal complexes, which led to the identification of several ribosome assembly intermediates and a large number of novel ribosome assembly factors. In this mini-review, we summarize some of the discoveries that have been made in the field of ribosome biogenesis in the past 30 years and highlight some key aspects about what remains to be learned. PMID- 15120993 TI - The SMN complex. AB - The survival of motor neurons (SMN) protein is the product of the disease determining gene of the neurodegenerative disorder spinal muscular atrophy (SMA). SMN is part of a stable multiprotein complex that is found in all metazoan cells in the cytoplasm and in nuclear Gems. The SMN complex contains, in addition to SMN, at least six other proteins, named Gemins2-7, and plays an essential role in the assembly of the spliceosomal small nuclear ribonucleoproteins (snRNPs). Through its binding to specific sequences in the snRNAs, the SMN complex surveys the correct identity of the target RNAs and facilitates snRNP assembly. Based on its ability to interact with several other protein and RNA components of cellular RNPs, it is likely that the SMN complex functions as an assemblyosome in the formation of diverse RNP particles, some of which may be of particular importance to the motor neuron. A detailed understanding of the cellular roles of the SMN complex may help the development of therapeutic strategies for this neurodegenerative disease. PMID- 15120994 TI - The XY body: a specialized meiotic chromatin domain. AB - The sex chromosomes of mammalian spermatocytes form a specialized nuclear territory known as the XY body, where both transcription and homologous recombination are restricted. The array of proteins assembled into the XY body is typical of heterochromatin. This special subnuclear domain is in distinct contrast to the autosomal domain of the spermatocyte nucleus, where both homologous recombination and transcription occur. The special features of the XY body might reflect absence of homology between the sex chromosomes, rather than any form of dosage compensation, and may also serve to mark parental origin of the paternal X chromosome. PMID- 15120995 TI - Spatial genome organization. AB - The linear sequence of genomes exists within the three-dimensional space of the cell nucleus. The spatial arrangement of genes and chromosomes within the interphase nucleus is nonrandom and gives rise to specific patterns. While recent work has begun to describe some of the positioning patterns of chromosomes and gene loci, the structural constraints that are responsible for nonrandom positioning and the relevance of spatial genome organization for genome expression are unclear. Here we discuss potential functional consequences of spatial genome organization and we speculate on the possible molecular mechanisms of how genomes are organized within the space of the mammalian cell nucleus. PMID- 15120996 TI - Mismatch repair proteins, meiosis, and mice: understanding the complexities of mammalian meiosis. AB - Mammalian meiosis differs from that seen in lower eukaryotes in several respects, not least of which is the added complexity of dealing with chromosomal interactions across a much larger genome (12 MB over 16 chromosome pairs in Saccharomyces cerevisiae compared to 2500 MB over 19 autosome pairs in Mus musculus). Thus, the recombination machinery, while being highly conserved through eukaryotes, has evolved to accommodate such issues to preserve genome integrity and to ensure propagation of the species. One group of highly conserved meiotic regulators is the DNA mismatch repair protein family that, as their name implies, were first identified as proteins that act to repair DNA mismatches that arise primarily during DNA replication. Their function in ensuring chromosomal integrity has also translated into a critical role for this family in meiotic recombination in most sexually reproducing organisms. In mice, targeted deletion of certain family members results in severe consequences for meiotic progression and infertility. This review will focus on the studies involving these mutant mouse models, with occasional comparison to the function of these proteins in other organisms. PMID- 15120997 TI - The mechanism of sister chromatid cohesion. AB - Each of our cells inherit their genetic information in the form of chromosomes from a mother cell. In order that we obtain the full genetic complement, cells need to ensure that replicated chromosomes are accurately split and distributed during cell division. Mistakes in this process lead to aneuploidies, cells with supernumerous or missing chromosomes. Most aneuploid human embryos are not viable, and if they are, they develop severe birth defects. Aneuploidies later in human life are frequently found associated with the development of malignant cancer. DNA replication during S-phase is linked to segregation of the sister copies in mitosis by sister chromatid cohesion. A chromosomal protein complex, cohesin, holds replicated sister DNA strands together after their synthesis. This allows pairs of replication products to be recognised by the spindle apparatus in mitosis for segregation into opposite direction. At anaphase onset, cohesin is destroyed by a site-specific protease, separase. Here I review what we have learned about the molecular mechanism of sister chromatid cohesion. Cohesin forms a large proteinaceous ring that may hold sister chromatids by encircling and topological trapping. To understand how cohesin links newly synthesised replication products, biochemical assays to study the enzymology of cohesin will be required. PMID- 15120998 TI - Nuclear architecture and genome functioning in plants and animals: what can we learn from both? AB - In this short review, we combine information about large-scale chromosome and chromatin structure in the interphase nucleus from animals and plants in relation to transcription. We propose a rosette-like structure for interphase chromosomes in higher eukaryotes in which condensed heterochromatin domains act as organizing centers from which euchromatic loops sprout that contain the transcription units available to that particular cell. In the same context, we address the apparent discrepancy between the spatial distribution of transcriptionally active sites in the nucleus and our understanding of the linear organization of genes on the linear genome. It is likely that this paradox can be solved by taking into account the probably large number of genes for nonprotein-coding RNAs. PMID- 15120999 TI - The link between mRNA processing and transcription: communication works both ways. AB - Many pre-mRNA processing events including 5' end capping, splicing out introns, and 3' end maturation by cleavage or polyadenylation occur while the nascent RNA chain is being synthesized by RNA polymerase II. As a consequence of this arrangement, the physiological substrate for most processing factors is not a solitary pre-RNA but instead a ternary complex comprising a growing RNA chain spewing from the exit channel of an RNA polymerase II molecule as it speeds along a chromatin template at 1000-2000 bases/min. mRNA processing factors make protein protein contacts with elongating pol II in a complex we have dubbed the "mRNA factory," which carries out synthesis, processing, and packaging of the transcript. Recent studies have shown that the "mRNA factory" is a dynamic complex whose composition changes as it traverses the length of a gene. This complex is also the setting for a growing number of regulatory interactions, which influence the function of both the processing and transcription machineries. PMID- 15121000 TI - Simulation of five intracellular C2+-regulation mechanisms in response to voltage clamp pulses. AB - We report the simulation of five C2+ regulation mechanisms in response to voltage clamp pulses applied at the membrane. We obtain the currents generated and also record the C2+ concentration changes in the whole cell during and after the pulse. Our simulator includes models for the L-type voltage-dependent C2+ channels, the C2+ pump, the Na+/C2+ exchanger and a fast buffering system, along with diffusion assuming a spheric cell shape. PMID- 15121001 TI - Spectral analysis of internal carotid arterial Doppler signals using FFT, AR, MA, and ARMA methods. AB - In this study, Doppler signals recorded from internal carotid artery of 45 subjects were processed by PC-computer using classical (fast Fourier transform) and model-based (autoregressive, moving average, autoregressive moving average (ARMA) methods) methods. Power spectral density estimates of internal carotid arterial Doppler signals were obtained using these spectral analysis methods. The variations in the shape of the Doppler power spectra as a function of time were presented in the form of sonograms in order to determine the degree of internal carotid artery stenosis. These Doppler power spectra and sonograms were then used to compare the applied methods in terms of their frequency resolution and the impact on determining stenosis in internal carotid arteries. Based on the results, performance characteristics of the autoregressive and ARMA methods were found extremely valuable for spectral analysis of internal carotid arterial Doppler signals obtained from healthy subjects and unhealthy subjects having artery stenosis. PMID- 15121002 TI - Biomimetic model of skeletal muscle isometric contraction: I. an energetic viscoelastic model for the skeletal muscle isometric force twitch. AB - This paper describes a revision of the Hill-type muscle model so that it will describe the chemo-mechanical energy conversion process (energetic) and the internal-element stiffness variation (viscoelastic) during a skeletal muscle isometric force twitch contraction. The derivation of this energetic-viscoelastic model is described by a first-order linear ordinary differential equation with constant energetic and viscoelastic coefficients. The model has been implemented as part of a biomimetic model, which describes the excitation-contraction coupling necessary to drive the energetic-viscoelastic model. Finally, the energetic-viscoelastic model is validated by comparing its isometric force-time profile with that of various muscles reported in the literature. PMID- 15121003 TI - Biomimetic model of skeletal muscle isometric contraction: II. A phenomenological model of the skeletal muscle excitation-contraction coupling process. AB - This paper describes a new macroscopic, phenomenological model of the skeletal muscle excitation-contraction coupling process, as represented by four principal and consecutive compartments (biophysical, biochemical, and biomechanical phases) characteristic of isometric excitation-contraction coupling in mammalian skeletal muscle, and coupled by a system of simultaneous, first-order linear ordinary differential equations. The model is based upon biological compartmental transport kinetics and irreversible thermodynamic energy transformation, and represents a distinct improvement over other biomimetic models. The model was derived using physiological parameter data published in the literature, and validated using MATLAB R12. PMID- 15121004 TI - Spectral broadening of ophthalmic arterial Doppler signals using STFT and wavelet transform. AB - In this study, short-time Fourier transform (STFT) and wavelet transform (WT) were used for spectral analysis of ophthalmic arterial Doppler signals. Using these spectral analysis methods, the variations in the shape of the Doppler spectra as a function of time were presented in the form of sonograms in order to obtain medical information. These sonograms were then used to compare the applied methods in terms of their frequency resolution and the effects in determination of spectral broadening in the presence of ophthalmic artery stenosis. A qualitative improvement in the appearance of the sonograms obtained using the WT over the STFT was noticeable. Despite the qualitative improvement in the individual sonograms, no quantitative advantage in using the WT over the STFT for the determination of spectral broadening index was obtained due to the poorer variance of the wavelet transform-based spectral broadening index and the additional computational requirements of the wavelet transform. PMID- 15121005 TI - An information-theoretic approach to estimating ultrasound backscatter characteristics. AB - Analysis of backscatter in the ultrasound echo envelope, in conjunction with ultrasound B-scans, can provide important information for tissue characterization and pathology diagnosis. Statistical models have often proven useful in modeling backscatter. In this paper, an innovative approach to backscatter analysis based on generalized entropies and neural function approximation is presented. Entropy measures are shown to provide accurate estimates of scatterer density, regularity, and SNR of the amplitude distribution. Specific scattering distributions need not be assumed. Experimental results on ground truth envelopes show that generalized entropies can be used to accurately estimate backscatter properties. PMID- 15121006 TI - The attainment of peak bone mass: what is the relationship between muscle growth and bone growth? PMID- 15121007 TI - The 'muscle-bone unit' during the pubertal growth spurt. AB - Mechanostat theory postulates that developmental changes in bone strength are secondary to the increasing loads imposed by larger muscle forces. Therefore, the increase in muscle strength should precede the increase in bone strength. We tested this prediction using densitometric surrogate measures of muscle force (lean body mass, LBM) and bone strength (bone mineral content, BMC) in a study on 70 boys and 68 girls who were longitudinally examined during pubertal development. On the level of the total body, the peak in LBM accrual preceded the peak in BMC accretion by an average of 0.51 years in girls and by 0.36 years in boys. In the arms, the maximal increase in LBM was followed by arm peak BMC accrual after an interval of 0.71 years in girls and 0.63 years in boys. In the lower extremities, the maximal increase in LBM was followed by peak BMC accrual after an interval of 0.22 years in girls and 0.48 years in boys. A multiple regression model revealed that total body peak LBM velocity, but not peak height velocity and sex, was independently associated with total body peak BMC velocity (r(2) = 0.50; P < 0.001). Similarly, arm and leg peak LBM velocity, but not peak height velocity and sex, were independently associated with arm and leg peak BMC velocity, respectively (r(2) = 0.61 for arms, r(2) = 0.41 for legs; P < 0.001 in both cases). These results are compatible with the view that bone development is driven by muscle development, although the data do not exclude the hypothesis that the two processes are independently determined by genetic mechanisms. PMID- 15121008 TI - Normal mineralization and nanostructure of sclerotic bone in mice overexpressing Fra-1. AB - Increased bone mass due to elevated number of active osteoblasts has been reported for transgenic mice overexpressing the transcription factor Fra-1. To explore the potential of the anabolic action of Fra-1 in treatment of osteoporosis, we examined the integrity of bone matrix generated in Fra-1 transgenic mice. Femora from Fra-1 transgenic (Fra-1 tg) and wild-type littermates were analyzed for bone mineralization density distribution (BMDD) and nanostructure using quantitative backscattered electron imaging (qBEI) and scanning small angle X-ray scattering (scanning-SAXS), respectively. For comparison, we studied mice lacking c-Fos (Fos-/-), which develop osteopetrosis because of the absence of osteoclasts. Morphometrical analysis of metaphyseal spongiosa revealed an up to 5-fold increase in bone volume for Fra-1 transgenic compared to wild type. BMDD indicated a transient lower mineralization of bone for Fra-1 transgenic at 5 and 8 weeks, which became comparable to that of wild type mice by 8 months. The homogeneity of mineralization was not altered in the Fra-1 transgenic mice at any ages examined. However, it was strikingly reduced in Fos-/- due to an abundance of hypermineralized cartilage. The bone nanostructure did not show abnormalities in Fra-1 transgenic or Fos-/-. These results provide a rationale for the development of therapeutic applications involving Fra-1-induced bone formation. PMID- 15121009 TI - The degree of mineralization is a determinant of bone strength: a study on human calcanei. AB - Strength of bones depends on bone matrix volume (BMV), bone microarchitecture, and also on the degree of mineralization of bone (DMB). We have recently shown in osteoporotic patients treated with alendronate that fracture risk decreased and bone mineral density increased with a parallel increase of the DMB due to prolonged secondary mineralization but without modifications of BMV or bone microarchitecture. DMB and strength were both measured at the tissue level in calcaneus bone samples taken at autopsy from 20 subjects (aged 78 +/- 8 years, 8 women, 12 men) who died suddenly without apparent bone disease. DMB parameters measured on microradiographs (mean DMB, distribution of DMB, most frequent maximum DMB value, and width at half maximum, an index reflecting the homogeneity of DMB) were compared with those reported in iliac cancellous bone samples of 43 human bones. Histomorphometric measurements of microarchitectural parameters (TbTh, TbN, and TbSp) were also measured. Compression tests were performed on contiguous samples of the same calcaneus on a universal screw-driven machine (Schenck RSA 250). A 5000-N load cell (TME, F 501 TC) measured the compressive load. The displacement was measured directly on the sample using a specific displacement transducer developed by the <> The apparent Young's modulus (E), the maximal strength (sigma(max)), and the work (W) until failure were measured. In human cancellous bone tissue, mean DMB (+/- SD) was higher in calcaneus (1.135 +/- 0.147 g/cm(3)) than in iliac crest (1.098 +/- 0.077 g/cm(3)). The mean most frequent maximum DMB values (mean DMB freq. max.) were 1.118 +/- 0.175 g/cm(3) in calcaneus and 1.108 +/- 0.095 g/cm(3) in iliac samples, and DMB was more heterogeneous in calcaneus than in iliac samples (mean width at half maximum were 0.270 +/- 0.127 versus 0.227 +/- 0.056 g/cm(3), respectively). Compression tests revealed significant positive linear correlations between DMB and both elastic modulus (r(2) = 0.69) and maximal strength (r(2) = 0.69). Correlations with DMB persisted (P < 0.003) even after adjustment for both calcified bone volume, for the Young's modulus (E), the maximal strength (sigma(max)) (r(2) = 0.44 and 0.41, respectively), and microarchitectural parameters (0.50 < r(2) < 0.56, P < 0.001). The same results were obtained with the work to fracture (W) (0.23 < r(2) < 0.46, P < 0.045). We conclude that the more the cancellous tissue was mineralized, the higher was its stiffness and compressive strength. This may explain the increase in bone strength when DMB is modified in a physiological range without necessary changes of BMV and bone microarchitecture. The impact of such modifications on fracture risk and the therapeutic implications of these data remain to be analyzed. PMID- 15121010 TI - On the origin of the toughness of mineralized tissue: microcracking or crack bridging? AB - Two major mechanisms that could potentially be responsible for toughening in mineralized tissues, such as bone and dentin, have been identified-microcracking and crack bridging. While evidence has been reported for both mechanisms, there has been no consensus thus far on which mechanism plays the dominant role in toughening these materials. In the present study, we seek to present definitive experimental evidence supporting crack bridging, rather than microcracking, as the most significant mechanism of toughening in cortical bone and dentin. In vitro fracture toughness experiments were conducted to measure the variation of the fracture resistance with crack extension [resistance-curve (R-curve) behavior] for both materials with special attention paid to changes in the sample compliance. Because these two toughening mechanisms induce opposite effects on the sample compliance, such experiments allow for the definitive determination of the dominant toughening mechanism, which in the present study was found to be crack bridging for microstructurally large crack sizes. The results of this work are of relevance from the perspective of developing a micromechanistic framework for understanding fracture behavior of mineralized tissue and in predicting failure in vivo. PMID- 15121011 TI - Osteopontin is a negative regulator of proliferation and differentiation in MC3T3 E1 pre-osteoblastic cells. AB - Osteopontin (OPN) is an important mediator of bone remodeling. However, the role of OPN in the process of bone formation is not fully understood. In previous studies, we have shown that MC3T3-E1 pre-osteoblastic cells at higher passage number exhibited weakened osteogenic capacity and elevated OPN mRNA expression. In this work, we investigated the role of OPN on proliferation and differentiation of low-passage MC3T3-E1 cells by studying stable cell lines overexpressing either OPN mRNA or its antisense RNA. Overexpression was verified by both Northern and Western blot analyses. Overexpression of OPN markedly inhibited proliferation as determined by daily cell counts, while overexpression of antisense RNA stimulated cellular proliferation. We also examined the effect of OPN level on BMP-2-induced alkaline phosphatase activity. Overexpression of OPN inhibited BMP-2 responsiveness while overexpression of antisense RNA enhanced the effect of BMP-2 on alkaline phosphatase activity. Increased OPN expression also caused decreases in expression of osteocalcin and bone sialoproteins while a reduction of OPN level caused the opposite. Furthermore, endogenous OPN expression in response to BMP-2 exhibited a biphasic pattern, that is, it was initially inhibited and then enhanced by the treatment of BMP-2, indicating that OPN might function as a negative feedback regulator for osteoblastic differentiation. Finally, overexpression of OPN inhibited mineral deposition. In contrast, overexpression of antisense RNA enhanced mineral deposition. These results indicate that OPN is a negative regulator of proliferation and differentiation in MC3T3-E1 cells. PMID- 15121012 TI - Primary murine limb bud mesenchymal cells in long-term culture complete chondrocyte differentiation: TGF-beta delays hypertrophy and PGE2 inhibits terminal differentiation. AB - In vitro models of endochondral bone formation using both primary and immortalized cells have provided insight regarding factors and signaling pathways involved in chondrocyte maturation and endochondral bone formation. However, primary murine cell culture models of chondrocyte differentiation have not been established but have enormous potential due to the possible use of cells from transgenic and knockout animals. Here, we show that stage E11.5 embryonic murine limb bud mesenchymal stem cells in micromass cell culture progress through the stages of chondrogenesis, chondrocyte hypertrophy, terminal differentiation, and matrix calcification. This cell culture system recapitulated the sequential expression of genes that characterize chondrocyte differentiation, including Sox9, col2, colX, MMP13, VEGF, and osteocalcin. TGF-beta treatment for up to 21 days markedly delayed the rate of chondrocyte maturation and inhibited matrix calcification and its related gene expression. In TGF-beta-treated cultures, the hypertrophic and terminal differentiation markers colX, VEGF, MMP13, and osteocalcin were reduced or absent. PGE2 had minimal effects on chondrocyte hypertrophy but delayed terminal differentiation and matrix calcification. Thus, primary murine mesenchymal cells sequentially differentiate through the various stages of chondrocyte maturation and establish a model whereby the role of specific signaling molecules can be examined in cells derived from transgenic or knockout mice. PMID- 15121013 TI - Wnt signaling inhibits osteogenic differentiation of human mesenchymal stem cells. AB - Human mesenchymal stem cells (hMSCs) from the bone marrow represent a potential source of pluripotent cells for autologous bone tissue engineering. We previously discovered that over activation of the Wnt signal transduction pathway by either lithium or Wnt3A stimulates hMSC proliferation while retaining pluripotency. Release of Wnt3A or lithium from porous calcium phosphate scaffolds, which we use for bone tissue engineering, could provide a mitogenic stimulus to implanted hMSCs. To define the proper release profile, we first assessed the effect of Wnt over activation on osteogenic differentiation of hMSCs. Here, we report that both lithium and Wnt3A strongly inhibit dexamethasone-induced expression of the osteogenic marker alkaline phosphatase (ALP). Moreover, lithium partly inhibited mineralization of hMSCs whereas Wnt3A completely blocked it. Time course analysis during osteogenic differentiation revealed that 4 days of Wnt3A exposure before the onset of mineralization is sufficient to block mineralization completely. Gene expression profiling in Wnt3A and lithium-exposed hMSCs showed that many osteogenic and chondrogenic markers, normally expressed in proliferating hMSCs, are downregulated upon Wnt stimulation. We conclude that Wnt signaling inhibits dexamethasone-induced osteogenesis in hMSCs. In future studies, we will try to limit release of lithium or Wnt3A from calcium phosphate scaffolds to the proliferative phase of osteogenesis. PMID- 15121014 TI - Stimulation of cAMP production and cyclooxygenase-2 by prostaglandin E(2) and selective prostaglandin receptor agonists in murine osteoblastic cells. AB - Prostaglandins (PGs), particularly PGE(2), can stimulate bone resorption and formation and auto-amplify their effects by inducing cyclooxygenase (COX)-2. We examined the role of different PG receptors in stimulating cAMP production and COX-2 expression in murine calvarial osteoblasts. Cells were obtained from PGE(2) receptor (EP2R and EP4R) wild-type and knockout (KO) mice and from mice transgenic for the COX-2 promoter fused to a luciferase reporter. We analyzed effects of selective agonists, EP2A and EP4A, for EP2R and EP4R, which mediate the increase in cAMP in response to PGE(2). We also tested agonists for other PGE(2) receptors (EP1A and EP3A) and for prostacyclin (IPA), prostaglandin D(2) (DPA), thromboxane (TPA), and prostaglandin F(2alpha) (FPA) receptors. PGE(2) and EP2A were the most effective stimulators of cAMP production. EP4A, IPA, and DPA produced smaller responses, and EP1A, EP3A, FPA, and TPA were ineffective. In EP2R KO cells, cAMP responses to PGE(2) were reduced by 80%, and responses to EP2A were abrogated. In EP4R KO cells, cAMP responses to PGE(2) and EP2A showed a small reduction, while the response to EP4A was abrogated. Pretreatment with PGE(2), EP2A, or EP4A down-regulated the subsequent response to the respective ligands. COX-2 induction was measured by increased luciferase activity and mRNA expression. PGE(2) was the most effective agonist; EP2A and another selective EP2R agonist, butaprost, showed similar efficacy, and EP4A was less effective. EP2A and EP4A effects on luciferase activity were additive, and effects of the combination were similar to PGE(2) itself. IPA, TPA, and DPA produced 2- to 6 fold increases in COX-2 expression. FPA was a weak agonist, while EP1A and EP3A were inactive. Treatment with specific inhibitors indicated that PGE(2), EP2A, and EP4A induced COX-2 expression largely through protein kinase A (PKA). We conclude that the PG induction of COX-2 in this system generally paralleled effects on cAMP production and was mediated predominantly via the PKA pathway. PMID- 15121016 TI - Predicting duration of Ilizarov frame treatment for tibial lengthening. AB - AIM: To predict duration of treatment for tibial lengthening, as a function of target length increase. METHOD: Review of case notes and radiographs of patients undergoing Ilizarov method of tibial distraction osteogenesis. RESULTS: In the 27 cases suitable for analysis, the median age was 29.9 years (13.3 to 72.6 years), and the median distraction was 3.5 cm (1.0 to 7.2 cm). A positive linear relationship (r = 0.7) was demonstrated between the distraction length (x, in cm) and the frame duration (y, in days), according to the equation y = 54x + 94. A negative hyperbolic relationship was also demonstrated between rate of bone healing and distraction length (r = 0.6), with distractions over 3.0 cm healing significantly faster than shorter ones (P < 0.03). PMID- 15121015 TI - Binding studies of [(18)F]-fluoride and polyphosphonates radiolabelled with [(111)In], [(99m)Tc], [(153)Sm], and [(188)Re] on bone compartments: a new model for the pre vivo evaluation of bone seekers? AB - INTRODUCTION: Although the first polyphosphonates were already introduced in the early 1970s, mechanisms involved in uptake still remain speculative. The present work aimed to establish a new method to rate the influence of various factors on the uptake and to evaluate new bone-seekers on these bone compartments. METHODS: Radioactive-labelled diphosphonates and [(18)F]-fluoride were added to a vial containing hydroxyapatite (HA), collagen, or amorphous calcium phosphate (ACP) in 3 ml of Hanks' Balanced Salt Solution (HBSS). After incubation, these suspensions were filtered, the radioactivity was measured in the gamma-counter, and the percentage of irreversibly bound radioactivity was calculated. RESULTS: Kinetic experiments revealed uptake increase over time for [(99m)Tc]-MDP and [(18)F] fluoride on various amounts of matrix. After 120 min, static studies on HA yielded: [(99m)Tc]-EDTMP < [(188)Re]-/Re-EDTMP < [(99m)Tc]-/11 microl Re-EDTMP < [(99m)Tc]-/In-EDTMP < [(99m)Tc]-/15 microl Re-EDTMP < nca [(188)Re]-EDTMP < [(111)In]-/Re-EDTMP < [(111)In]-EDTMP < [(111)In]-/In-EDTMP < [(99m)Tc]-DPD < [(99m)Tc]-/80 microl Re-EDTMP < [(99m)Tc]-EDTMP "boiled" < [(99m)Tc]-/150 microl Re-EDTMP < [(153)Sm]-EDTMP < [(99m)Tc]-/11 microl Re-EDTMP "boiled" < [(18)F] ions < [(99m)Tc]-MDP. Collagen showed very low uptake. Reincubation experiments suggest that bone tracers are irreversibly bound. CONCLUSION: The presented method is rapid and feasible to examine the adsorption of radioactive-labelled substances on bone components. Correlations between our findings and published in vivo data support the application as a simple model. PMID- 15121017 TI - The role of angiogenesis in a murine tibial model of distraction osteogenesis. AB - Distraction osteogenesis (DO) is one of the most dramatic in vivo applications of mechanical stimulation as a means of inducing bone regeneration. A simple and reproducible murine model of tibia distraction osteogenesis was developed using a monolateral fixator. Bone formation was assessed histologically over a 35-day time course. The steady state expression of a broad family of angiogenesis associated genes was assessed by microarray hybridization analyses over the same time course, while the immediate gene response that was induced during each cycle of distraction was assessed at 30 min and 8 h after the first and last rounds of activation of the fixator. Distraction osteogenesis promoted new bone formation primarily through an intramembranous process with maximal osteogenesis during the active distraction period. Histological analysis also showed that dense cortical bone continued to be formed, during the consolidation phase, for 2 weeks after distraction ended. The analysis of steady state mRNA expression levels over the time course of DO showed that VEGF-A and neuropilin, an alternate receptor for VEGF-A, both angiopoietin (Ang) 1 and 2 factors, and the Ang receptor Tie2 were the critical angiogenic factors during DO. A key transcriptional regulator of many of the angiogenic factors, hypoxia-induced factor1alpha (Hif-1a), the FGF binding protein pleiotropin/OSF1, and multiple MMP(s), were also induced during the active distraction period. Examination of the expression of angiogenic factors that were induced after each cycle of activation, demonstrated that Hif 1a, Nrp1, and VEGF-A were all cyclically induced after each increment of distraction. These results suggest that these factors are early mediators that are produced by distraction and contribute toward the processes that promote bone formation. These experiments represent the first step in defining the molecular mechanisms that regulate skeletal regeneration and the functional relationship between angiogenesis and osteogenesis during distraction osteogenesis. PMID- 15121018 TI - Local delivery of growth factors from coated titanium plates increases osteotomy healing in rats. AB - Different methods for the stabilization of long bone fractures are used in clinic. Besides the development of further stabilization devices, the use of new materials, the modification of the surfaces, and the local application of stimulating factors for enhancement of healing are from great interest. Previous studies successfully used a biodegradable poly(d,l-lactide) coating as a local drug delivery system of growth factors from intramedullary (IM) implants to enhance fracture healing. In this study, we developed a new rat model (n = 60) for plate osteosynthesis and used a plate for stabilization and as a local drug delivery system for the growth factors IGF-I and TGF-beta1. A four-hole titanium plate was used for stabilization of a 0.6-mm osteotomy gap of the femur. The space between the inner holes was coated with 50 microg IGF-I and 10 microg TGF beta1 incorporated in the poly(d,l-lactide) coating or with the coating alone. After 42 days, biomechanical tests and histomorphological analyses were performed to investigate osteotomy healing. Radiologically small differences were detectable between the groups. The biomechanical torsional testing revealed a significantly higher maximum load of the osteotomized femura after treatment with growth factors compared to the uncoated group. In the histomorphometric analyses measuring the callus composition, a significantly higher percentage of mineralized tissue in the osteotomy callus was assessed in the growth factor treated group compared to the uncoated. In conclusion, the local application of IGF-I and TGF-beta1 from a biodegradable coating enhances the osteotomy healing as shown in the biomechanical testing and the histomorphometry. Bioactive plates could be used in clinic for fracture stabilization and for local and controlled application of growth factors to stimulate bone healing. PMID- 15121019 TI - Relationship between the duration of paralysis and bone structure: a pQCT study of spinal cord injured individuals. AB - The aim of the present study was to describe bone loss of the separate compartments of trabecular and cortical bone, as well as changes in bone geometry of a large number of spinal cord injured (SCI) individuals. Eighty-nine motor complete spinal cord injured men (24 tetraplegics and 65 paraplegics) with a duration of paralysis of between 2 months and 50 years were included in the study. Distal epiphyses and midshafts of the femur, tibia, and radius were measured by peripheral quantitative computed tomography. The same measurements were performed in a reference group of 21 healthy able-bodied men of the same age range. In the femur and tibia, bone mass, total and trabecular bone mineral density (BMDtot and BMDtrab, respectively) of the epiphyses, as well as bone mass and cortical cross-sectional area of the diaphyses, showed an exponential decrease with time after injury in the spinal cord injured subjects. The decreasing bone parameters reached new steady states after 3-8 years, depending on the parameter. Bone mass loss in the epiphyses was approximately 50% in the femur and 60% in the tibia, while the shafts lost only approximately 35% in the femur and 25% in the tibia. In the epiphyses, bone mass was lost by reducing BMD, while in the shaft bone mass was lost by reducing cortical wall thickness, a process achieved by endosteal resorption advancing at a rate of about 0.25 mm/year within the first 5-7 years after injury. Except for a slight transient decrease in cortical BMD of the femoral and tibial shaft during the first 5 years after the spinal cord lesion, cortical BMD of the spinal cord injured subjects was found to be at reference values. Bone parameters of the radial epiphysis in paraplegic subjects showed no deficits compared to the reference group. Furthermore, a trend for an increased radial shaft diameter suggests periosteal apposition as a consequence of increased loading of the arms. PMID- 15121021 TI - Insufficiently dosed intravenous ibandronate injections are associated with suboptimal antifracture efficacy in postmenopausal osteoporosis. AB - Less frequent bisphosphonate dosing in women with postmenopausal osteoporosis has the potential to promote therapy adherence through improved convenience. Ibandronate is a highly potent nitrogen-containing bisphosphonate, proven to significantly increase vertebral and nonvertebral bone mineral density (BMD) when administered as a convenient intravenous injection. A recent double-blind, placebo-controlled, randomized phase III study explored the antifracture efficacy and safety of 1 and 0.5 mg iv ibandronate injections, given once every 3 months, in 2862 women (55-76 years) with postmenopausal osteoporosis [one to four prevalent vertebral fractures and lumbar spine (L1-L4) BMD T score of less than 2.0 and greater than -5.0 in >or=1 vertebra]. All participants received daily vitamin D (400 IU) and calcium (500 mg) supplementation. The primary endpoint was the incidence of new morphometric vertebral fractures after 3 years. However, although a consistent trend toward a reduction in the incidence of new morphometric vertebral fracture was observed in the active treatment arms compared with placebo (9.2% vs. 8.7% vs. 10.7% in the 1 mg, 0.5 mg and placebo groups, respectively), as well as in the incidence of nonvertebral and hip fractures, the magnitude of fracture reduction was suboptimal and was insufficient to achieve statistical significance. At the studied doses, intravenous ibandronate injections also produced dose-dependent, but comparatively small, increases in lumbar spine BMD (4.0% and 2.9%, respectively) and decreases in biochemical markers of bone resorption and formation, relative to placebo. Optimal fracture efficacy likely requires more substantial increases in BMD and more pronounced suppression of bone turnover. In light of the clear dose-response relationship observed in this and other studies, this is likely to be achieved with higher intravenous doses of ibandronate. The results of a recent phase II/III study (Intermittent Regimen Intravenous Ibandronate Study: the IRIS study) provide support for this hypothesis. PMID- 15121020 TI - Efficacy and safety of ibandronate given by intravenous injection once every 3 months. AB - Oral bisphosphonates are established therapeutics for postmenopausal osteoporosis. Alternative, simplified dosing regimens that improve tolerability and promote convenience may be advantageous. Ibandronate is a highly potent, nitrogen-containing bisphosphonate that can be administered as a convenient intravenous (i.v.) injection (over 15-30 s) in schedules featuring extended between-dose intervals. In a recent fracture prevention study, 1 and 0.5 mg i.v. ibandronate injections, given once every 3 months, were shown to dose-dependently increase lumbar spine and hip bone mineral density (BMD) and decrease biochemical markers of bone turnover in women with postmenopausal osteoporosis, but the overall magnitude of efficacy provided by both doses was suboptimal. In the present study (Intermittent Regimen intravenous Ibandronate Study: the IRIS study), the dose-response relationship with intermittent intravenous ibandronate injections was further evaluated in 520 postmenopausal osteoporotic women (aged 55-75 years, time since menopause >or= 5 years, lumbar spine [L1-L4] BMD T score < -2.5). At enrolment, participants were randomized to receive either 2 mg (n = 261) or 1 mg (n = 131) ibandronate or placebo (n = 128) intravenous injections, given once every 3 months. After 1 year, ibandronate therapy produced substantial and dose-dependent increases in lumbar spine and hip BMD, and decreases in biochemical markers of bone turnover, with the 2 mg dose providing significantly greater efficacy than the 1 mg dose. Most notably, lumbar spine BMD increased by 5.0% and 2.8% in the 2 and 1 mg groups, respectively, and decreased by 0.04% in the placebo group. Furthermore, total hip BMD increased by 2.9%, 2.2%, and 0.6%, respectively. Serum and urinary CTX, reflecting bone resorption, were decreased by 62.5% and 61%, respectively, with the 2 mg dose, and by 43.5% and 42%, respectively, with the 1 mg dose. Intravenous ibandronate was well tolerated with a similar incidence of adverse events to placebo. Importantly, no indicators of renal toxicity were reported. In summary, the 2 mg ibandronate regimen provides significantly greater BMD increases and significantly greater suppression of bone resorption markers than the 1 mg dose used in this study and in the previous fracture prevention study. Ongoing studies aim to further establish the efficacy and convenience of intermittent intravenous ibandronate injections in postmenopausal osteoporosis. PMID- 15121022 TI - Bone mineral density in diabetic children and adolescents: a follow-up study. AB - OBJECTIVE: To establish whether T1DM can affect bone mineral density (BMD) in children and adolescents. RESEARCH DESIGN AND METHODS: We performed a cross sectional and longitudinal study of 57 diabetic children and adolescents and 57 normal controls. Total body and lumbar BMD and bone mineral content (BMC) were assessed by DXA (Lunar DPX) and volumetric transformation was calculated using the Katzman formula for total body BMD (BMAD) and using the Kroger formula for Lumbar BMD (L2L4BMDvol). BMC, BMAD, BMDspine, and L2L4BMDvol were adjusted for confounding factors such as age, gender, BMI, height, weight, and pubertal stage. RESULTS: BMDspine in the control group increased by 0.006 (g/cm(2))/year; while in the 39 diabetic patients longitudinally studied, it dropped by 0.006 (g/cm(2))/year during a follow-up period of 51 +/- 27 months. The average time spent weekly doing physical activity resulted in T1DM group directly correlated to BCM (P < 0.001) and inversely correlated with BMDspine (P < 0.05) and L2L4BMDvol (P < 0.01). L2L4BMDvol resulted significantly correlated with previous BMD spine (R = 0.63; P < 0.0001) and BMC evaluation (R = 0.42; P < 0.01) but not with BMAD. A second lumbar DXA evaluation performed in 38 patients after 1.00 +/- 0.16 years confirmed a small but significant decrease of 1.6% per year in L2L4BMDvol. The percentage of variation of L2L4BMDvol between the two evaluations was not correlated with the level of metabolic control, insulin requirement, and duration of the disease. Patients with complications showed similar L2L4BMDvol to patients without complications. CONCLUSIONS: Diabetic children and adolescents show a slight negative pattern of spine mineralization, which does not depend on metabolic control and microvascular complications. PMID- 15121023 TI - Resolution of severe, adolescent-onset hypophosphatemic rickets following resection of an FGF-23-producing tumour of the distal ulna. AB - Oncogenic hypophosphatemic osteomalacia (OHO) is an uncommon hypophosphatemic syndrome characterized by bone pain, proximal muscle weakness and rickets. It has been postulated that OHO results from overproduction of a humoral phosphaturic factor by an occult tumour. Recently, some OHO tumours have been shown to elaborate fibroblast growth factor-23 (FGF-23), which causes renal phosphate wasting when administered to mice. The purpose of this study was to undertake detailed investigations to confirm the diagnosis of OHO in a pediatric patient and to document the biochemical, radiographic and bone histological phenotype before and after tumour removal. We describe an 11-year-old, previously healthy girl with significant pain and functional disability associated with hypophosphatemic rickets. Circulating 1,25-(OH)(2) vitamin D was very low (14 pM; N: 40-140) while the FGF-23 serum level was markedly elevated [359.5 reference units (RU)/ml, N: 33-105]. An iliac bone biopsy revealed severe osteomalacia, but periosteocytic lesions, as are typical for X-linked hypophosphatemic rickets, were not seen. Sequence analyses of the PHEX and FGF23 genes were normal. A radiographic skeletal survey revealed a small exostosis of the left, distal ulnar metaphysis. A tumour was subsequently removed from this site and the pathology was consistent with benign, fibro-osseous tissue. Serum FGF-23 was normal when measured at 7 h post-operatively, while serum phosphate reached the low-normal range at 16 days following surgery. An iliac bone biopsy taken 5 months after the operation showed improvement, but not yet resolution, of the osteomalacia. Biochemical parameters of bone and mineral metabolism suggested that complete resolution of the osteomalacia was not achieved until 12 months following surgery. One year after tumour removal, the patient was pain-free and had resumed a normal level of activity. The rapid normalization of FGF-23 levels following removal of a benign tumour and the subsequent improvement in the biochemical and histological parameters of bone and mineral metabolism suggest that FGF-23 played a key role in this girl's disease. PMID- 15121024 TI - On Stolken and Kinney (bone 2003;33(4):494-504). PMID- 15121026 TI - Can confounding with fat-derived endogenous free estradiol explain the inverse correlation of bone mineral density with adiponectin? PMID- 15121028 TI - Cardiovascular risk--through the ages. PMID- 15121029 TI - Do pregnancy complications and CVD share common antecedents? AB - Considerable data link low birth weight, due to intrauterine growth restriction, to increased offspring risk of vascular disease in later adult life. This is considered to be the result, in part, of programming through fetal nutrition. These data support the hypothesis that pregnancy outcome in terms of birth weight is linked to the infant's subsequent health. In contrast, much less attention has been focused on the relationship between adverse pregnancy outcomes, such as pre eclampsia, gestational diabetes, pre-term delivery and intrauterine growth restriction, and the mother's subsequent health. Interesting data have accumulated linking the maternal vascular, metabolic and inflammatory complications of pregnancy to an increased risk of vascular disease in later life (Table 1). This paper reviews the emerging evidence to support this fascinating concept, addresses potential mechanisms and discusses potential clinical implications. PMID- 15121030 TI - Impact of age on cardiovascular risk: implications for cardiovascular disease management. AB - Cardiovascular disease (CVD) represents a major global healthcare problem. The prevalence of this condition increases with age. As many countries around the world are experiencing an increase in the proportion of elderly people in the population, this raises serious issues for cardiac and cerebrovascular disease prevention and management. A wealth of data has established smoking, dyslipidemia, hypertension and type 2 diabetes as major risk factors for cardiac and cerebrovascular events. This article reviews the evidence that links these metabolic risk factors with an increased risk of complications, and assesses the data concerning how risk changes with age. This review also focuses on how these conditions can be optimally managed and whether treatment outcomes are affected by age. The current status of research is assessed and issues which remain to be resolved are highlighted. PMID- 15121031 TI - Insights from apoB: from better diagnosis & therapy to the Medusa Hypothesis. AB - This article will review six of the most important insights that have come from the measurement of apolipoprotein B (apoB). Amongst these are critical clinical advances, which include better definition of those at high risk of vascular disease and better definition of the adequacy of statin therapy. There are also major advances in our understanding of the fundamental processes that interact to cause vascular disease. These include a more precise understanding of the determinants of lipoprotein levels and lipoprotein composition in plasma and, even more importantly, an appreciation of the pivotal role excess fatty acid intake and impaired fatty acid trapping by adipose tissue play in the generation of the complex risk profile that is so common in patients with coronary disease. We have designated the constellation of hypertriglyceridemic (hyperTg) hyperapoB, dysglycemia, and elevated levels of markers of prothrombosis and inflammation- all of which are traceable to adipose tissue dysfunction--as the Medusa Hypothesis. The Medusa Hypothesis puts forward a common pathophysiology for the complex and biologically diverse array of factors that have been associated with vascular disease. PMID- 15121032 TI - HDL: a recipe for longevity. AB - The concentration of high density lipoprotein cholesterol (HDL-C) has been found to be a powerful negative predictor of premature coronary heart disease (CHD) and stroke in human prospective population studies. Evidence of the protective properties of HDLs has also been documented in the elderly and their offspring. HDLs mediate several functions that provide an insight into their potential anti atherogenic mechanisms. Intervention strategies to prevent CHD have generally focused on lowering low-density lipoprotein cholesterol (LDL-C). However, several lifestyle and pharmacological interventions have the capacity to raise the level of HDL-C. As data accumulate on the protective role of HDLs, there is growing support for interventions that act to raise HDL-C concentrations. PMID- 15121033 TI - Gene therapy and CVD: how near are we? AB - Critical for success of any gene therapy approach is the efficient packaging, effective cell specific delivery and nuclear translocation of the nucleic acid with minimal toxicity. Delivery systems utilizing a wide variety of viral vectors have traditionally been used to modify genomic DNA. However, drawbacks to the viral vectors include difficulties in large-scale production, potential contamination by wild-type viral particles and immunogenicity. Thus, efficient non-viral delivery of both plasmids for transgene expression and short oligonucleotides for modulating cellular functions has been developed. Gene therapy is now a consideration in the treatment of certain inherited and acquired genetic disorders associated with cardiovascular disease (CVD). Furthermore, many other cardiovascular conditions are potential targets for gene therapy, and advances in knowledge will increase the ability to link specific genes to a disease, resulting in the identification of further targets. With improvements in delivery and targeting, gene therapy is likely to substantially augment established and emerging therapies in reducing the global burden of cardiovascular disease. PMID- 15121034 TI - Primary prevention in rheumatology: the importance of hyperuricemia. AB - Hyperuricemia (HU) is present in 5-30% of the general population, although the prevalence is higher among some ethnic groups and seems to be increasing worldwide. Classically, chronic HU has been considered a risk factor for gout or lithiasis and is associated with alcoholism, obesity, hypertension, dyslipidemia, hyperglycemia/diabetes mellitus, renal failure and intake of certain drugs. HU is also associated with cardiovascular diseases such as hypertension, vascular disease, pre-eclampsia, pulmonary arterial hypertension, stroke, heart failure, ischemic heart disease and also metabolic syndrome, renal disease and increased mortality. It is uncertain if these associations are dependent or not, especially cardiovascular and renal diseases. Patients with chronic HU and also those with gout require both medical investigation for associated diseases or drugs as well as nutritional counseling and life-style changes. HU should alert physicians to possible complications. PMID- 15121035 TI - Different approaches to understanding autoimmune rheumatic diseases: the neuroimmunoendocrine system. AB - Rheumatic autoimmune diseases are characterized by dysregulation of the immune response that leads to inflammation, pain, disease and stiffness and have been shown to have differences in disease pattern, depending on the gender and age of an individual. The majority of these conditions predominantly affect females of all species and also show increased severity of disease in female animal models. In addition to the gender differences in disease development, persons are often more susceptible at specific stages of life. This review will discuss some of the data indicating age and gender differences in development of these diseases and will review hormonal and other factors that may contribute to disease expression and severity. PMID- 15121036 TI - New approaches to understanding the impact of musculoskeletal conditions. AB - The approval of the new international classification of functioning, disability and health (ICF; formerly ICIDH) (WHO, 2001a) by the World Health Assembly (WHA) in May 2001 is a landmark event for medicine and society. With the ICF, patients' functioning, with its components body functions and structures, activities and participation, became a central perspective in medicine. The objective of this paper is to outline how the ICF can serve as a new global language of functioning and health and hence become a new approach for our understanding of the impact of musculoskeletal conditions. PMID- 15121037 TI - New insights into culture driven disorders. AB - Rheumatologists frequently encounter patients whose illnesses lack face-value; that is, they lack the typical objective features of pathology that rheumatologists traditionally rely on for diagnosis and developing effective treatment approaches: namely fibromyalgia, chronic fatigue syndrome, Gulf War syndrome, chronic whiplash, chronic low back pain, etc. In this article, we examine this group of illnesses as culture-driven disorders to emphasize the central importance of various societal constraints in the ultimate presentation of patients with these illnesses. We will examine them by first understanding the purpose they serve, the underlying factors that compel societal institutions to sanctify these disorders as diseases, and how research is beginning to examine the behaviour that captures and packages these symptoms to produce their clinical presentation. With this research understanding, rheumatologists may be able to offer patients more useful action plans, but likely changes in societal approaches to the expressions of distress and changes in disability and compensation systems will also be required. PMID- 15121038 TI - A biomechanical approach to musculoskeletal disease. AB - The importance of biomechanical factors in musculoskeletal disease may be appreciated in the clinical characteristics and pathophysiology of joint degeneration in osteoarthritis. The biomechanical characteristics of the neuromuscular system are integral in determining the function and stability of the synovial joint and in mediating the biochemical structure of articular cartilage. Alterations in the neuromuscular system including abnormal gait and dynamic joint loading patterns as well as muscle strength and proprioception deficits have been independently studied and associated with osteoarthritis. These factors have close functional and physiological interactions. Nevertheless, specific relations between muscle strength and proprioception and their independent contributions to dynamic joint loading are not yet clear. Targeted interventions to moderate these factors may, in the future, be a therapeutic option for the management of osteoarthritis. PMID- 15121039 TI - Tissue repair in rheumatoid arthritis: challenges and opportunities in the face of a systemic inflammatory disease. AB - Rheumatoid arthritis (RA) is a systemic inflammatory disease that can elicit a variable disease course, can influence a variable number of joints, and can exhibit a variable response to treatment. All of these factors contribute to the degree and extent of damage to joint components, as well as the potential for repair of other injured joint tissues/components. Furthermore, some of the RA treatments/drugs themselves can influence repair and injury responses, as well as the outcome of surgical interventions for advanced disease. However, as treatments and interventions become more sophisticated and successful in patient populations, the opportunity to initiate the repair/replacement of the damaged joint tissues also becomes more of a reality. This review will address the current clinical findings in the literature, and then discuss the issues and opportunities to initiate repair of damaged or injured joint tissues in order to restore joint function. These include growth factors, gene therapy, and bioengineered tissues, alone or in combination to augment endogenous repair or replace tissue damaged beyond such repair capabilities. PMID- 15121040 TI - An economic approach to health care. AB - Economic analyses have the potential to put all of the positive and negative outcomes of an intervention into perspective to aid decision making. The quality of the data upon which the analysis is based has an impact on the resulting quality of the analysis itself. Analysis of cost-effectiveness requires the input of many types of data, and where data are not available, assumptions must be made. There are many instances where the analysis may go wrong, and it is important to remain cognizant of these. The critical parts of the analysis, which have also been identified in quality assessment tools, include the following: design of the study question, sources of probability estimates and cost data, sensitivity analysis, and the interpretation of results. If the readers are able to identify the assumptions of the analysis they are better equipped to judge the validity. We have reviewed economic analyses relating to two hot economic topics in rheumatology. These are the cost-effectiveness of cyclooxygenase-2 (COX-2) inhibitors for 'arthritis' and cost-effectiveness of anti-tumor necrosis factor alpha (anti-TNF) agents for rheumatoid arthritis (RA). The results of the COX-2 analyses vary by review. Some show cost savings, while others calculate a significant cost in order to achieve any change in quality of life. Given the unanswered questions that still exist, it seems reasonable to conclude that COX-2 inhibitors may be cost effective when used in patients at a high risk of GI complications. Unanswered questions remain regarding the concomitant use of low dose ASA and proton pump inhibitors and how they may affect the results of these economic analyses. The cost-effectiveness of anti-TNF agents has not been explored in as much detail as that of the COX-2 agents. Two studies have presented cost-effectiveness models that include a hypothetical biologic agent. Two economic analyses report on the cost-effectiveness of etanercept compared with traditional disease-modifying anti-rheumatic drugs (DMARDs) in methotrexate resistant and methotrexate-naive patients with RA. Both the analyses show that etanercept has a cost-effectiveness ratio of around 40,000 US dollars for every patient who achieves an American College of Rheumatology 20% improvement score (ACR 20) within a 6-month period. A cost-utility analysis was published regarding the use of infliximab in methotrexate resistant RA. It showed a cost-utility ratio of 3400:34,000 Euro per quality adjusted life year (QALY) gained, depending on the country evaluated (Sweden and the UK, respectively). An important finding in all three studies was that indirect costs dominate costs in RA; therefore, they should be included in all future analyses of this disease. PMID- 15121041 TI - New insights into sexual functioning and fertility in rheumatic diseases. AB - Sexuality is an often neglected area of quality of life in patients with rheumatic disease. Manifestations and symptoms of disease can impair sexual functioning, but this can be much improved by adequate intervention and counseling. Fertility is in general not reduced in rheumatic diseases, however, the time taken to achieve a pregnancy is often increased. An increased rate of pregnancy loss is observed in systemic lupus erythematosus and the antiphospholipid syndrome contributing to a reduced family size. Autoantibodies are present in most of the rheumatic diseases and can interfere with fertilization, implantation, embryonic development and placental function. Active disease disturbs the hypothalamic-pituitary-axis, giving rise to periods of gonadal dysfunction. Toxic effects of immunosuppressive drugs can induce transient or permanent gonadal failure in women and men. PMID- 15121042 TI - Pharmacogenetics of disease-modifying anti-rheumatic drugs. AB - The outcome of treatment with disease-modifying anti-rheumatic drugs (DMARDs) in rheumatoid arthritis (RA) patients is considerably variable and is also unpredictable. It would be useful clinically if physicians were able to predict responses to DMARDs prior to their administration. One possible cause of differences in efficacy and adverse drug reactions is genetic variation in how individuals metabolize drugs. Based on pharmacogenetics, tailor-made drug therapy, also called personalized drug therapy or individual drug therapy, will be possible with analysis of genetic polymorphism, such as single nucleotide polymorphism (SNP), and analysis of haplotype and diplotype configuration. Several studies of the correlation between the genetic polymorphism of enzymes metabolizing several DMARDs and efficacy or adverse drug reactions have already been reported, suggesting that pharmacogenetics will be applicable to the treatment of RA in the near future. PMID- 15121044 TI - Ezrin, a key component in tumor metastasis. AB - Identification of the key regulatory molecules in metastasis is crucial for understanding tumor dissemination and for the development of novel interventions. The recent identification of ezrin as a necessary component in the metastasis of osteosarcoma and rhabdomyosarcoma is, therefore, an important advance. Ezrin has been implicated in many roles, for example, as a conduit for signals between metastasis-associated cell-surface molecules and signal transduction components. This suggests that ezrin and, potentially, other members of the ERM (ezrin radixin-moesin) family have key roles in the coordination of signals and cellular complexes that are required for the successful metastasis of these and other malignancies. PMID- 15121045 TI - Quality and quantity: new strategies to improve immunotherapy of cancer. AB - Adoptive immunotherapy is a promising approach for the treatment of infectious diseases and cancer. Several lines of research are currently focusing on the development of different technologies to facilitate the induction and expansion of antigen-specific T cells. Here, we discuss two current articles that affect the field of adoptive immunotherapy. One article describes the engineering of artificial antigen-presenting cells, which promise to replace the cumbersome dendritic-cell approach for the in vitro generation of large numbers of antigen specific T cells. The second development is a description of a new technique for the detection of functionally active antigen-specific T cells, which will enhance the ability to control the quality of the T cells to be used in adoptive immunotherapy. Together, these exciting findings will advance the field of immunotherapy. PMID- 15121046 TI - Darwinism and pharmacogenomics: from 'one treatment fits all' to 'selection of the richest'? PMID- 15121047 TI - Vesicular stomatitis virus: re-inventing the bullet. AB - As our understanding of the molecular aspects of human disease increases, it is becoming possible to create designer therapeutics that are exquisitely targeted and have greater efficacy and fewer side effects. One class of targeted biological agents that has benefited from recent advances in molecular biology is designer viruses. Vesicular stomatitis virus (VSV) is normally relatively innocuous but can be engineered to target cancer cells or to stimulate immunity against diseases such as AIDS or influenza. Strains of VSV that induce or direct the production of interferon are superior to wild-type strains of the virus for inducing oncolysis. These strains might also make better vaccine vectors. In this review, some of the features that make VSV an excellent platform for the development of a range of viral therapeutics are discussed. PMID- 15121048 TI - Schistosome transcriptomes: new insights into the parasite and schistosomiasis. AB - Schistosomiasis is one of the most serious parasitic diseases. More than 250 million people are infected with schistosomes in the tropics or subtropics. The parasitic flukes have some unique biological features: dioecism, complex life cycles, mechanisms to avoid host immune responses, and an apparent reliance on host endocrine and immune signals to complete their development, maturation and egg production. Recently, a large dataset of expressed sequence tags (ESTs) were generated from Schistosoma japonicum and Schistosoma mansoni, from which numerous novel genes were identified. The transcriptome analyses provide the basis for a comprehensive understanding of the molecular mechanisms involved in schistosome nutrition and metabolism, host-dependent development and maturation, immune evasion and invertebrate evolution. In addition, new potential vaccine candidates and drug targets have been predicted. PMID- 15121049 TI - Mutations in CD46, a complement regulatory protein, predispose to atypical HUS. AB - Membrane cofactor protein (MCP, CD46) is a widely expressed transmembrane complement regulator. As does the soluble regulator factor H, it inhibits complement activation by inactivating the C3b that is deposited on target membranes. Factor H mutations have been described in 15-30% of patients with atypical haemolytic uraemic syndrome (HUS). Recent studies have identified mutations in the MCP gene in four families. In one, a heterozygous deletion resulted in the intracellular retention of the mutant protein. In another, a different heterozygous deletion led to a premature stop codon and the loss of the C-terminus. In the other two, a substitution (S206P) resulted in cell-surface expression but inefficient inactivation of surface-bound C3b. These findings provide further evidence that complement dysregulation predisposes to the development of HUS. PMID- 15121050 TI - Recent advances in understanding serotonin regulation of cardiovascular function. AB - Serotonin is an important neurohormonal factor that has been implicated in cardiovascular function. It can regulate vascular tone, act directly on cardiomyocytes and stimulate chemosensitive nerves in the heart. Cardiovascular dysfunction is observed when serotonin signaling is altered or when variation in serotonin concentration occurs. Recent studies have provided evidence that, in the absence of peripheral serotonin synthesis, blood serotonin (which is almost exclusively stored in platelets) is markedly reduced, and that this drop leads to heart failure. This implies that the level of circulating serotonin is a key factor in maintaining normal cardiovascular activity. These findings offer new prospects for the use of serotonin in therapies for cardiovascular diseases. PMID- 15121051 TI - Use of cell-permeable peptides to prevent neuronal degeneration. AB - The loss of neurons is a hallmark of neurodegenerative disorders and evidence suggests that this occurs through an apoptotic mechanism. Following an insult, neuronal cells activate signal transduction pathways that lead to cell death and the establishment of the pathological state. The mechanisms underlying the cell death response involve protein kinases, which phosphorylate many substrates and culminate in changes in gene expression. Traditionally, attempts at blocking such signaling targeted the phosphorylation of the substrates. However, preventing the interaction between two proteins using specific peptides might block the function of key mediators in signaling cascades. A cell-permeable peptide designed specifically to inhibit c-Jun N-terminal kinase action proved successful in in vivo models of neuronal degeneration following ischemia. Here, the recent findings that highlight the potential of this approach for therapeutic application are reviewed. PMID- 15121052 TI - Design and manufacture of a polyvinyl alcohol (PVA) cryogel tri-leaflet heart valve prosthesis. AB - Although current artificial heart valves are life sustaining medical devices, improvements are still necessary to address deficiencies. Bioprosthetic valves have a compromised fatigue life, while mechanical valves have better durability but are prone to thromboembolic complications. A novel, one-piece, tricuspid valve, consisting of leaflets, stent and sewing ring, made entirely from the hydrogel, polyvinyl alcohol cryogel (PVA-C), has been developed and demonstrated. This valve has three thin leaflets attached to a cylindrical stent. In order to approximate the complex shape of the surface of the natural heart valve leaflets, two different geometries have been proposed: revolution about an axis of a hyperboloid shape and revolution about an axis of an arc subtending (joining) two straight lines. The parameters of both geometries were examined based on a compromise between avoiding sharp curvature of leaflets and minimization of the central opening of the valve when closed. The revolution of an arc subtending two straight lines was selected as the preferred geometry since it has the benefit of a smaller central opening when the value of the maximum curvature for the leaflets is the same for each valve geometry. A cavity mold has been designed and constructed to form the PVA-C heart valve. The three leaflets were formed and integrated into the stent and sewing ring in a single process. Prototype heart valves were manufactured in the mold from a solution of PVA and water, by controlled freezing and thawing cycles. PMID- 15121053 TI - Effect of medication in Parkinson's disease: a wavelet analysis of EMG signals. AB - The improvements in the motor ability in patients with Parkinson's disease due to antiparkinsonian medication is well-known and widely documented. Recent results, based both on kinematic parameters and standard electromyographic (EMG) signal analysis, clearly indicated that the medication reduced, as expected, the clinical signs of Parkinson's disease, but did not restore agonist burst duration modulation with distance in elbow flexion movements. The main aim of the present work is to shed more light on this medication effect using a wavelet analysis approach on multiple EMG signals recorded both on shoulder and elbow muscles in ballistic or rapid movements. The wavelet cross-correlation information allows us to evidence some important quantitative features of the EMG signals due to medication. PMID- 15121054 TI - Spectral pattern complexity analysis and the quantification of voice normality in healthy and radiotherapy patient groups. AB - Vocal fold functionality may alter in response to direct radiotherapy or indirectly by perturbation of the hypothalamic-pituitary axis. Perceptual assessment of voice quality is difficult to summarise in a single, reliable figure of normality and normality itself is undefined. In this study spectral analysis of vocal fold vibration, based on impedance variations measured across the larynx using an electro-glottogram, is used to build a single parameter description of standard vowel phonation in the normal male population. Patient data and perceptual assessment are then compared to this standard. The spectral pattern of the vowel/i/ electro-glottogram time series is analysed using approximate entropy after dynamic fundamental-harmonic frequency normalisation. The approximate entropy provides a single estimate of the spectral pattern complexity. A cohort of 89 normal males formed two statistically distinct groups, G1, with strong spectral pattern and high complexity 0.338 (+/-0.036), and G2 with a weak spectral pattern and low complexity 0.175 (+/-0.049). Membership ratio G1:G2 was 2:1. A cohort of 30 male larynx cancer cases were analysed approximately 3-6 months after irradiation, and three male prophylactic cranial irradiation cases some years after treatment. Two-thirds of patients had G2 or lower levels of complexity. The lower G2 complexity level appears to be the subjective, as well as the objective, threshold for voice normality. PMID- 15121055 TI - Detection of single and clustered microcalcifications in mammograms using fractals models and neural networks. AB - Microcalcifications (microCas) are often early signs of breast cancer. However, detecting them is a difficult visual task and recognizing malignant lesions is a complex diagnostic problem. In recent years, several research groups have been working to develop computer-aided diagnosis (CAD) systems for X-ray mammography. In this paper, we propose a method to detect and classify microcalcifications. In order to discover the presence of microCas clusters, particular attention is paid to the analysis of the spatial arrangement of detected lesions. A fractal model has been used to describe the mammographic image, thus, allowing the use of a matched filtering stage to enhance microcalcifications against the background. A region growing algorithm, coupled with a neural classifier, detects existing lesions. Subsequently, a second fractal model is used to analyze their spatial arrangement so that the presence of microcalcification clusters can be detected and classified. Reported results indicate that fractal models provide an adequate framework for medical image processing; consequently high correct classification rates are achieved. PMID- 15121056 TI - Wavelet packet transform for R-R interval variability. AB - INTRODUCTION: Wavelet transform is used for time-frequency analysis. Recently, discrete wavelet transform (DWT) has been used to analyze R-R interval or heart rate variability. However, we hypothesized that wavelet packet transform (WPT) is a better way to analyze such variability. In the present study, we compared resolution of frequency band and amplitude, which are used for analysis of the variability, with DWT and WPT, followed by Hilbert transform. METHODS: A chirp signal which covers all frequency bands used for R-R interval variability was employed as a simulated signal. Levels 1-6 of DWT and level 3 of WPT were used for signal analysis. Amplitudes of the gained signal were evaluated with Hilbert transform. Differences in error of the gained amplitude from expected amplitude between CWT and DWT for low-frequency (LF) and high-frequency (HF) components were compared. To evaluate time-dependent changes in R-R interval variability, head-up tilt (HUT) was employed as an orthostatic challenge. RESULTS: Errors for both HF and LF, derived from the simulated signal with WPT, were significantly smaller than those of DWT. With HUT, time dependent changes in LF, HF, and LF/HF were observed. DISCUSSION: Although DWT is a valuable method for time-frequency analysis, WPT is a more appropriate method to utilize wavelet transform due to the equivalent resolution of the gained frequency band. WPT for time-frequency analysis improves analysis of time-dependent changes in R-R interval variability. PMID- 15121057 TI - Improvement and automation of artificial neural networks to estimate medical outcomes. AB - The lengthy process of manually optimizing a feedforward backpropagation artificial neural network (ANN) provided the incentive to develop an automated system that could fine-tune the network parameters without user supervision. A new stopping criterion was introduced--the logarithmic-sensitivity index--that manages a good balance between sensitivity and specificity of the output classification. The automated network automatically monitored the classification performance to determine when was the best time to stop training-after no improvement in the performance measure (either highest correct classification rate, lowest mean squared error or highest log-sensitivity index value) occurred in the subsequent 500 epochs. Experiments were performed on three medical databases: an adult intensive care unit, a neonatal intensive care unit and a coronary surgery patient database. The optimal network parameter settings found by the automated system were similar to those found manually. The results showed that the automated networks performed equally well or better than the manually optimized ANNs, and the best classification performance was achieved using the log-sensitivity index as a stopping criterion. PMID- 15121058 TI - Updating a 3-D vertebral body finite element model using 2-D images. AB - In osteoporotic patients, vertebral strength is often evaluated in the clinical setting using bone densitometry methods, such as dual energy X-ray absorptiometry. Finite element models based on computed tomography (CT) have been shown to outperform such methods in predicting fracture strength, but repetitive use of CT scans may be impractical due to cost, availability, and radiation exposure. We propose a method of generating a vertebral model at an initial time point which can subsequently be updated using several digital radiographs by using an algebraic reconstruction technique (ART) to find the three-dimensional density distribution. The effectiveness of the algorithm was measured by comparison of the error of the reconstructed model to the error incurred by using the outdated model. Application of the ART was able to reduce density errors from 30% to under 7% and to reduce errors in calculated vertebral stiffness from 125% to under 10%. This preliminary study demonstrates that the method is valid and could potentially improve fracture risk diagnostics drastically. PMID- 15121059 TI - A cam-displacement transducer device for measuring small two-degree of freedom inter-component motion in a prosthesis. AB - An electromechanical system is described that comprises a cam and two linear displacement transducers for measuring two-degree of freedom. This is aimed at quantifying the extent and pattern of both transverse rotation and longitudinal translation that takes place during amputee gait at what is sometimes termed a "torque-shock absorbing adapter" located between the prosthetic socket and foot. Both types of motion are measured independently by mounting the cam around the adapter and the two transducers orthogonal to one another with their spring loaded plungers moving separately against the cam's perimeter and distal, flat face. The cam geometry should be an arc of a spiral and not a circle for the angular displacement measurement to be linear in response to rotation. Over a range of +/-18 degrees for transverse rotation the maximum deviation of the ideal cam geometry away from an arc of a circle is less than +/-0.5% of the transducers' 10 mm stroke. The transducer's linearity is +/-2% and it could therefore be assumed that this would be the dominant source of error. The theoretical predictions are supported by the calibration results, in which the pattern of errors that are found in the transducers' calibration can also be seen in the calibration of the cam. PMID- 15121060 TI - The kinetic model and simulation of blood coagulation--the kinetic influence of activated protein C. AB - The paper described a limited part of the coagulation pathway, and in particular the inhibitory effects of activated protein C in the context of thrombin production. This is a computational modeling study with various assumption made of kinetic rates laws and their summation. The level of complexity and assumed parameters makes conclusions uncertain. However, an interesting outcome is that kinetic reaction rates may show oscillation behavior under particular, high levels of protein C feedback inhibition. The model would defy quantitative practical use, but could have predictive value as a qualitative descriptor of coagulation. PMID- 15121061 TI - Bilateral symmetry of biomechanical properties in mouse femora. AB - Bone healing and remodeling are commonly examined in animal models by comparing one femur (experimental) to the contralateral femur (control) with the assumption that they are identical with respect to their biomechanical properties. While past studies have characterized the symmetry in geometrical properties in many types of animal bones, few studies have compared the symmetry in the biomechanical properties. The purpose of this study was to determine whether there is symmetry in the mechanical properties of mouse femora. Strain gauges were attached to the posterior surface of the femora of C57BL/6 mice, parallel to the long axis of the bone. The femora were mechanically tested in cantilever bending while strain values were recorded. Moments of inertia, cortical areas, and moduli of elasticity were determined from strains and cross-sectional properties. Mouse femora demonstrated an average strain difference of 0.4% in tension and 1.4% in compression. Elastic moduli differed by 6.6% and 0.9% in tension and compression, respectively, and failure strength differed by an average of 2.0%. Statistical analysis showed there were no significant differences in strain, modulus, or failure load values for the mice, indicating mechanical and geometrical symmetry of mouse femora in cantilever bending. PMID- 15121062 TI - A multi-station dynamic-culture force monitor system to study cell mechanobiology. AB - To study mechanobiological responses of cells, a dynamic-culture force monitor (D CFM) system has been developed. The D-CFM extends our previous work to measure contractile forces of a cell-populated collagen gel (CPCG) using a cantilever beam with semiconductor strain gauges. Linear actuators are used in the system and are computer controlled using a LabVIEW interface to independently apply precise motion waveforms to multiple CPCGs. The feasibility tests showed that the new system can detect the differences in force patterns resulting from different motion waveforms imparted to the CPCG. This new system will facilitate the study of the effects of dynamic mechanical loading on cells, remodeling of extracellular matrix, and cell-matrix interactions in vitro. PMID- 15121063 TI - Intramedullary femoral nails: one or two lag screws? A preliminary study. PMID- 15121065 TI - From trials to guidelines to clinical practice: the need for improvement. PMID- 15121066 TI - ICD-implantation guidelines versus clinical practice: a prospective study of out of-hospital cardiac arrest survivors. AB - AIMS: The aim of this study was prospectively to compare clinical practice of implantable cardioverter defibrillator (ICD) use with current guidelines in out of-hospital cardiac arrest (OHCA) survivors. METHODS AND RESULTS: From January 2000 till March 2002, 70 consecutive patients (pts) discharged from 15 hospitals after OHCA, with ventricular fibrillation (VF) as initial rhythm were included. Documentation of diagnosis, left ventricular function, ischaemia, electrophysiological studies (EPS), and decisions regarding ICD implantation were obtained from medical records. An expert committee compared these data with current guidelines. According to these guidelines 18 pts (26%) had an ICD indication and received an ICD while 37 pts (53%) had no indication and did not receive an ICD. In 13 pts without acute myocardial infarction insufficient diagnostic procedures were performed to permit a decision on ICD indication, hence no ICD was implanted. Two pts had an ICD indication but did not receive an ICD. During the follow-up with duration of 25 months (range 12-38 months), two sudden deaths occurred in the group of pts without an ICD. Of the pts with an ICD, 4 pts (22%) were reported to have received one or more shocks for VT/VF. CONCLUSION: In at least 21% of OHCA survivors, insufficient diagnostic procedures concerning the indication for ICD implantation were performed or no ICD was implanted when indicated, despite clear guidelines. In particular, there was no proof of ischaemia prior to revascularization and no confirmation of the absence of ischaemia and EPS thereafter. Clinicians should be guided better in evaluating pts after OHCA concerning the indication for ICD implantation, especially when a transient of reversible condition was present or when treatment was sufficiently established safely to refrain from ICD implantation. PMID- 15121067 TI - Primary hyperparathyroidism and arrhythmic storm in a patient with an implantable cardioverter defibrillator for primary prevention of sudden death. AB - A case of a patient with an automatic cardioverter defibrillator (ICD) and recurrent ventricular arrhythmic storms related to primary hyperparathyroidism and hypercalcaemia is reported: medical therapy was ineffective and only surgical resection of the parathyroid adenoma resolved this complex clinical condition. PMID- 15121068 TI - Defaecation syncope: new light on an old problem. PMID- 15121070 TI - Efficacy of tilt training in the treatment of neurally mediated syncope. A randomized study. AB - Recurrent neurally mediated syncope represents a common clinical event and a therapeutic challenge. Recently tilt training has been proposed for the treatment of recurrent neurally mediated syncope. To evaluate the efficacy of tilt training in preventing tilt-induced syncope and its feasibility, this controlled, randomized study was undertaken. Sixty-eight consenting patients (25 males and 43 females, mean age 40 +/- 19) with recurrent neurally mediated syncope and 2 consecutive positive nitroglycerin-potentiated head-up tilt tests were randomized to tilt training (35 patients) or no treatment (controls, 33 patients). The tilt training programme consisted of daily 30-min sessions of upright standing against a vertical wall 6 days a week for at least 3 weeks, until a reevaluation tilt test (3 patients of both groups dropped out). On this third head-up tilt test, 19 (59%) of 32 tilt trained patients and 18 (60%) of 30 controls still had a positive test. Treated patients performed a mean number of 15 +/- 7 sessions (median 16) and only 11 patients (34%) did all the programmed sessions. Only 1 patient (3%) discontinued treatment because of intolerance, while all other patients did not perform tilt training adequately, because of poor compliance. Thus, in our study tilt training was not effective in reducing tilt testing positivity rate in patients with neurally mediated syncope. Because of poor compliance, tilt training appears to be a feasible treatment only for highly motivated patients, but not for the majority of patients with recurrent neurally mediated syncope. PMID- 15121069 TI - Abnormalities of sympathetic and parasympathetic autonomic function in subjects with defaecation syncope. AB - Defaecation syncope is defined as blacking out at, or around, the time of defaecation. It is associated with increased mortality; however, patients rarely voluntarily report symptoms. We have examined autonomic function in a cohort of patients with defaecation syncope. METHODS: We prospectively identified all subjects referred to our unit with symptoms of defaecation syncope or presyncope on direct questioning. All subjects had autonomic function tests using beat to beat blood pressure measurement synchronized with R-R interval allowing real time assessment of autonomic function. RESULTS: Seven patients were identified who presented with defaecation syncope. Compared with age and sex matched controls, subjects had abnormalities of both sympathetic and parasympathetic autonomic function consistent with mild-moderate autonomic failure. On specific intervention syncope stopped in all subjects: 3 had culprit medication withdrawn, 3 received medication to increase blood pressure and 1 in whom cardioinhibition was demonstrated improved with permanent pacemaker insertion. Two subjects who had colonoscopy had profound haemodynamic changes during the procedure associated with syncope. CONCLUSIONS: Symptoms of syncope on defaecation are associated with autonomic failure. With appropriate therapeutic intervention our subjects all improved. PMID- 15121071 TI - Superior vena cava syndrome and syncope in an implantable cardioverter defibrillator recipient. AB - We describe the case of a recipient of an implantable cardioverter defibrillator with multiple syncopal episodes due both to superior vena cava obstruction and electrical instability. These complications occurred in the presence of two transvenous implantable cardioverter defibrillator leads. The patient has been managed conservatively with anticoagulants and new antiarrhythmic drugs with improvement in both his clinical problems. PMID- 15121072 TI - Radiofrequency catheter ablation of atrioventricular nodal reentrant tachycardia in children aided by the LocaLisa mapping system. AB - AIMS: In young patients, slow pathway ablation for treatment of atrioventricular nodal reentrant tachycardia (AVNRT) carries a small but definite risk of permanent AV block. The aim was to assess the efficacy of slow pathway ablation aided by the LocaLisa mapping system. PATIENTS AND METHODS: Radiofrequency (RF) modification of the slow AV nodal pathway was performed in 26 children < 19 years of age (median age 9.8 years, range 3-18.9). Three measures to limit the risk of AV block were applied: (1) use of LocaLisa, a non-fluoroscopic mapping system, to determine and mark the location of the AV node/His bundle axis, and monitor ablation catheter position, (2) continuous atrial stimulation during RF delivery to monitor AV conduction, and (3) gradual increase of RF power during RF ablation. RESULTS: AVNRT was rendered non-inducible in all patients. Dual AV physiology was abolished in 24/26 patients; 2 patients had single atrial echoes at the end of the procedure. At follow-up, AVNRT recurred in 3 patients (including the above 2), necessitating a second procedure. The median number of RF applications was 4 (3-8); median fluoroscopy time was 16 (7-33)min. One patient developed transient second-degree AV block, with full recovery within 6 weeks of the procedure. CONCLUSIONS: RF modification of the slow AV nodal pathway in children can be safely accomplished, achieving the ideal end-point of abolishing dual AV physiology, aided by use of the LocaLisa mapping system. PMID- 15121073 TI - Catheter ablation of atrial tachycardia due to recipient-to-donor transatrial conduction after orthotopic heart transplantation. AB - Atrial arrhythmias occur frequently after orthotopic heart transplantation. In some cases, they may depend on atrio-atrial conduction across the surgical suture line. We report an unusual case of a heart transplant recipient who had recipient atypical atrial flutter with 2:1 transatrial conduction and a resulting tachycardia in the donor atrium. Electroanatomical mapping identified the site of transatrial conduction and guided successful catheter ablation. PMID- 15121074 TI - Spontaneous transition of four different types of supraventricular tachycardias in one patient. PMID- 15121076 TI - Temporal changes in atrial refractoriness following DC cardioversion of persistent atrial fibrillation in man. AB - AIMS: Studies have demonstrated shortening of the atrial effective refractory period (ERP) after episodes of atrial fibrillation (AF). This is termed atrial remodelling. It is unclear whether restoration of SR after persistent AF in patients with a clinical substrate results in reversal of this shortening and whether this is maintained long term. METHODS AND RESULTS: The ERP was determined at mid-lateral right atrial wall (MLRA) and right atrial appendage (RAA) at 600 ms and 400 ms drive cycle lengths and at basic sinus cycle length in 81 patients with persistent AF immediately, 24 h and 2 weeks following external DC cardioversion. All atrially active drugs were stopped for at least 5 half lives. (1) Prolongation of the ERP was observed at both atrial sites and all cycle lengths up to 24 h post cardioversion (p < 0.0001). (2) However, between 24 h and 2 weeks a subsequent shortening occurred in the ERP returning it to near post cardioversion levels. (3) The ERP was significantly longer at 24 h post cardioversion in patients who remained in SR for 2 weeks or longer compared with those who reverted to AF. CONCLUSION: Prolongation of the atrial ERP occurred following restoration of SR in persistent AF patients but was not maintained and displayed a biphasic pattern such that by 2 weeks the ERP had returned to baseline values. Despite this finding, a longer ERP at 24 h post cardioversion was associated with maintenance of SR in the medium-term. PMID- 15121075 TI - Paroxysmal atrial fibrillation in male endurance athletes. A 9-year follow up. AB - AIMS: To study the course and circumstances of atrial fibrillation in endurance sports. BACKGROUND: Atrial fibrillation is the main cause of symptoms in athletes and frequently disturbs the performance during the practising of sport. The course of the arrhythmia is seldom reported in this population. METHODS: In 1993 and 2002 symptoms and clinical presentation of atrial fibrillation were evaluated in 30 well-trained athletes with a specially designed questionnaire. RESULTS: In 1993 paroxysmal atrial fibrillation was present in 30 male athletes at the mean age of 48.1 +/- 7.8, 3 (10%) of them also had paroxysmal atrial flutter. Three (10%) of the athletes died before 2002. In 2002 paroxysmal atrial fibrillation continued in 15 (50%) athletes, permanent atrial fibrillation emerged in 5 (17%) athletes and 7 (23%) of them showed no further atrial fibrillation. In 1993 paroxysms of atrial fibrillation started at a relatively low level of training intensity compared with the mean maximal training intensity of 11 +/- 7 versus 8 +/- 4 h/week (p < 0.05). The first attack of adrenergically induced paroxysmal atrial fibrillation was more often present in younger athletes (p < 0.005) and vagally induced paroxysmal atrial fibrillation was more apparent in older athletes (p < 0.05). In 10 (38%) of the athletes a familiar form of paroxysmal atrial fibrillation was present. CONCLUSION: A small proportion of the athletes (26%) was asymptomatic. Paroxysmal atrial fibrillation remained stable in half of the athletes whereas the arrhythmia changed into permanent atrial fibrillation in a minority of this population. PMID- 15121077 TI - Heart rate regularisation in patients with permanent atrial fibrillation implanted with a VVI(R) pacemaker. AB - Irregularity of ventricular cycles is a cause of haemodynamic impairment and symptoms in patients with atrial fibrillation (AF). AIM OF THE STUDY: Aim of the study was to determine the optimal pacing rate to stabilise ventricular cycle length at rest in patients with chronic AF, bradycardiac symptoms and VVI pacing. METHODS: The compensatory pause (CP) in AF, as defined by Langendorf, was used as a reference value in pacing the heart. The spontaneous mean heart rate (MHR) was assessed with the PM OFF. The CP was then calculated with the pacing rate programmed at 40 bpm. Four pacing rates were tested: rate of the CP (RCP), RCP + 5 bpm, RCP - 5 bpm and RCP - 10 bpm. RESULTS: RCP provided a good estimate of the MHR (r = 0.92). Pacing percentage (P%) was 24 +/- 15% at the pacing rate of RCP - 10 bpm, 39 +/- 19% at RCP - 5 bpm, 63 +/- 17% at RCP, and 79 +/- 19% at RCP + 5 bpm (p < 0.001). The corresponding HR modestly increased from 65 +/- 13 bpm to 66 +/- 13 bpm (p = NS), 68 +/- 13 bpm (p < 0.001) and 71 +/- 13 bpm (p < 0.001), respectively. CONCLUSION: The RCP estimates, during pacing, what the spontaneous MHR would be. Ventricular stimulation at the RCP causes a high P%, stabilising cardiac cycles with a modest increase in HR. PMID- 15121078 TI - Lead extraction for device related infections: a single-centre experience. AB - AIMS: We report a single-centre experience of extraction of infected pacing and ICD leads. METHODS: Retrospective study of consecutive lead extractions for infection. Lead extraction was by traction, or, if unsuccessful, a laser sheath was used. All procedures were performed in the operating room. RESULTS: Leads were extracted in 82 patients including 66 patients (80%) with infection occurring after replacement of the generator or revision of the leads. Previous treatment, without lead extraction, had failed in 51 patients (62%). Major complications (tamponade or haemothorax) occurred in 6 patients, 2 patients died despite emergency surgery. One patient succumbed to ongoing sepsis. Of the patients alive, a follow-up of at least 6 months (27 +/- 17 months) was available in 76 patients. All patients were cured; none had a recurrence. In 31 patients (41%) pacing was abandoned after lead extraction; all remained asymptomatic. CONCLUSION: Lead extraction is effective in curing pacemaker or ICD related infection, even after failed conservative therapy, but with a significant complication rate. The routine replacement of the generator should be reconsidered in patients in whom the indication for pacing is no longer valid, as the majority of infections occurred after revision of the system. PMID- 15121079 TI - Capture management efficacy in children and young adults with endocardial and unipolar epicardial systems. AB - AIMS: This prospective study characterized performance of the Kappa 700 Ventricular Capture Management trade mark (VCM) system for monitoring ventricular pacing threshold and adapting outputs in both endocardial and unipolar epicardial pacing systems in children and young adults. VCM bears cautionary labelling against use with epicardial leads since they have not been demonstrated appropriate for use with VCM. METHODS AND RESULTS: VCM was programmed in "Monitor Only" mode. Ventricular pacemaker thresholds were measured daily using VCM for a minimum of 2 months. Potential device longevities at nominal outputs (3.5 V, 1.0 ms) and at VCM-recommended outputs were compared. Thirty patients (median age 14.4 years (1-27 years); 15 epicardial/15 endocardial) completed the study. During the daily measurements, consistent undersensed evoked response occurred in 2 patients (Medtronic epicardial leads 4965). For the other 28 patients, programming VCM in "Adaptive" mode from implant would provide an additional 6.8 months (0-19 months) of battery life. CONCLUSION: Although not an IDE (Investigation Device Exemption) study, this study showed acceptable VCM performance in "Monitor Only" mode in 13/15 patients with unipolar epicardial leads. A 2-month "Monitor Only" observation period helps screen patients who might not benefit from VCM. VCM may provide substantial energy savings and extended battery life for children and young adults. PMID- 15121081 TI - Abnormal ascospore morphology in the bud mutant of Neurospora tetrasperma. AB - A recessive ascospore mutant of Neurospora tetrasperma, named bud, was isolated from a wild-collected heterokaryotic strain with four different nuclear components. bud segregates as a single mendelian gene. When bud is homozygous, meiosis is apparently normal but postmeiotic events are not. Abnormal orientation of spindles at the postmeiotic mitosis often results in failed pair-wise association of nuclei and their irregular distribution along the length of the ascus prior to spore delimitation. Consequently, many asci cut out more than four ascospores; some contain no nuclei while others contain more than two. The most dramatic effect of bud is on ascospore delimitation itself. Many ascospores are irregularly shaped and are often interconnected, because of incomplete spore delimitation. Ascospores also show one or two lobes or bud-like extensions of varying sizes. Over 75% of ascospores from bud x bud remain white or tan and are inviable. The interaction of bud with a dominant Eight-spore mutant (E) was examined in both heterozygous and homozygous crosses. When both bud and E are heterozygous, bud has no effect on ascospore delimitation or on the phenotype of E because bud is recessive; many asci produce 5-8 ascospores just as in E x E(+). And when bud is homozygous and E is heterozygous, ascospore delimitation is less affected than when E is absent. Moreover, when both bud and E are homozygous, the effect on ascospore development is less extreme than when E is homozygous singly. PMID- 15121082 TI - The interaction among evolutionary forces in the pathogenic fungus Mycosphaerella graminicola. AB - The population genetic dynamic of a species is driven by interactions among mutation, migration, drift, mating system, and selection, but it is rare to have sufficient empirical data to estimate values for all of these forces and to allow comparison of the relative magnitudes of these evolutionary forces. We combined data from a mark-release-recapture experiment, extensive population surveys, and computer simulations to evaluate interactions among these evolutionary forces in the pathogenic fungus Mycosphaerella graminicola. The results from these studies showed that, on average, the immigration rate was 0.027, the fraction of outcrossing individuals was 0.035, and the selection coefficient associated with immigrants was 0.106 each generation. We also estimated that effective population sizes for this fungus were larger than 24,000 and the mutation rate for the RFLP markers used in surveys and field experiments was approximately 4 x 10(-5). Computer simulations based on these estimates indicate that, on average, the global population of M. graminicola has reached equilibrium. Population genetic parameters including number of alleles, gene diversity, and population subdivision estimated from the computer simulations were surprisingly close to empirical estimates. Simulations also revealed that random drift is the major evolutionary force decreasing genetic variation in this fungus, followed by natural selection. The major force adding to genetic variation was mutation, followed by gene flow and sexual recombination. Gene flow played the leading role in decreasing population subdivision while natural selection was the major factor increasing population subdivision. PMID- 15121084 TI - Stage-specific cellular localisation of two hydrophobins during plant infection by the pathogenic fungus Cladosporium fulvum. AB - Hydrophobins are central to developmental processes of filamentous fungi. HCf-1 and HCf-6 are two of the six hydrophobins identified in the plant pathogenic fungus Cladosporium fulvum. We have fused the viral epitope V5 to HCf-1 and HCf 6, introduced the recombinant genes into C. fulvum strains that lack the two genes, and localised the tagged proteins by immunofluorescence microscopy. HCf 1(V5) is abundant on conidia and aerial structures formed in vitro and emerging from disease lesions on infected tomato plants. This is consistent with the proposed function of HCf-1 in aerial development and dissemination of conidia. HCf-6(V5) is secreted onto the growth substrate by the hyphae and during invasion of plant tissues, which suggests a function in adhesion and infection. This was not supported by the phenotypic analysis of DeltaHCf-6 strains. Hydrophobins may play distinct roles due to precisely regulated spatial localisation during infection-related development of C. fulvum. PMID- 15121085 TI - Differential expression of the chitin synthase genes of Aspergillus nidulans, chsA, chsB, and chsC, in response to developmental status and environmental factors. AB - To understand the role of the chitin synthase genes of Aspergillus nidulans, we analyzed the expression of chsA, chsB, and chsC both by Northern blotting and by a vital reporter system with sgfp encoding a modified version of green fluorescent protein, sGFP. chsA was expressed specifically during asexual differentiation, but not during either vegetative growth or sexual differentiation. The expression of chsB was ubiquitous throughout the fungal body and relatively independent of the change in developmental status of the cells. chsC was expressed moderately during sexual development as well as during the early phase of vegetative growth, but was expressed weakly in old vegetative mycelia and in asexual structures. Furthermore its expression was spatially differentiated, i.e., relatively strong in young cleistothecia and in mature ascospores, but negligible in Hulle cells. Osmostress caused by high concentrations (up to 1.2M) of KCl or NaCl stimulated the expression of chsA and chsC, but not that of chsB. Sodium acetate, especially at high concentration (3%), strongly enhanced the expression of all the three genes. Neither heat shock nor the sugar carbon sources tested (glucose, sucrose, or lactose) affected the expression of any of the three chitin synthase genes. PMID- 15121086 TI - Expression profile analysis of wild-type and fcc1 mutant strains of Fusarium verticillioides during fumonisin biosynthesis. AB - Fusarium verticillioides produces a group of mycotoxins known as fumonisins that are associated with a variety of mycotoxicoses in humans and animals. In this study, DNA microarrays were constructed with expressed sequence tags (ESTs) from F. verticillioides. To identify genes with patterns of expression similar to the fumonisin biosynthetic (FUM) genes, the microarray was probed with labeled cDNAs originating from a wild-type strain and a fcc1 mutant grown on maize and in a defined medium adjusted to either pH 3 or pH 8. The comparative analyses revealed differential expression of genes corresponding to 116 ESTs when the fungal strains were grown on maize. Under different pH conditions, 166 ESTs were differentially expressed, and 19 ESTs were identified that displayed expression patterns similar to the FUM ESTs. These results provide candidate genes with potential roles in fumonisin biosynthesis. PMID- 15121087 TI - Some red blood cell phenomena for the curious. PMID- 15121088 TI - A genome-based approach for the study of erythroid biology and disease. AB - The human genome contains all elements of the erythroid transcriptome with the exception of those genes encoded in mitochondrial DNA. The concept of a "genome based" approach for the study of erythroid biology and disease was envisioned in the mid-1990s as a logical offshoot of efforts to map the human genome. Although a completed human genome map was not expected until 2005 [Science 279 (1998) 23], the idea of creating a robust and retrievable description of erythroid gene activity was both encouraged and supported on the NIDDK Bethesda campus of the National Institutes of Health. The 5-10-year goals were organized into three parts. The initial project goal involved the collection of sequence data derived from mRNA expressed in developmentally staged erythroblasts. Those cells were isolated prospectively by flow cytometry monitoring of CD71 and glycophorin A expression patterns. Next, the sequence data was organized into a database and integrated with related information available in the public domain. The paramount goal of the project continues to be clinical application. As discussed in this paper, progress already made in each of these areas suggests that genome-based approaches will greatly facilitate future studies of erythroid biology and disease. PMID- 15121089 TI - The effect of mefloquine and volume-regulated anion channel inhibitors on induced transport in Plasmodium falciparum-infected human red blood cells. AB - The malaria parasite Plasmodium falciparum activates new permeation pathways (NPP) in the host cell membrane of infected human red blood cells (RBCs), which are permeable to anions, cations and a range of organic solutes. It has been suggested from inhibitor and substrate selectivity studies that the NPP may be identical to the volume-activated anion channel (VRAC) present in many mammalian cell types. Here we have tested several known inhibitors of VRAC on the transport of choline and lactate in malaria-infected human RBCs and on parasite growth. Mefloquine, tamoxifen and clomiphene were all without effect on malaria-induced transport at concentrations up to 10 microM and only mefloquine (IC(50) = 24 nM) and, to a lesser degree, clomiphene (IC(50) = 6.2 microM) inhibited parasite growth below this level. It is concluded that the antimalarial effect of mefloquine does not involve the inhibition of malaria-induced transport via the NPP and there is no evidence at present for VRAC and the NPP being identical. PMID- 15121083 TI - Genealogical concordance between the mating type locus and seven other nuclear genes supports formal recognition of nine phylogenetically distinct species within the Fusarium graminearum clade. AB - Species limits were investigated within the Fusarium graminearum clade (Fg clade) through phylogenetic analyses of DNA sequences from portions of 11 nuclear genes including the mating-type (MAT) locus. Nine phylogenetically distinct species were resolved within the Fg clade, and they all possess contiguous MAT1-1 and MAT1-2 idiomorphs consistent with a homothallic reproductive mode. In contrast, only one of the two MAT idiomorphs was found in five other species, four of which were putatively asexual, and the other was heterothallic. Molecular evolutionary analyses indicate the MAT genes are under strong purifying selection and that they are functionally constrained, even in species for which a sexual state is unknown. The phylogeny supports a monophyletic and apomorphic origin of homothallism within this clade. Morphological analyses demonstrate that a combination of conidial characters could be used to differentiate three species and three species pairs. Species rank is formally proposed for the eight unnamed species within the Fg clade using fixed nucleotide characters. PMID- 15121090 TI - Evidence for a mechanosensitive calcium influx into red cells. AB - Red cells exposed to glycophorin A reactive antibodies and lectins develop a non specific cation permeability. To determine if this might be due to the activation of a non-selective mechanosensitive channel we have subjected red cells, loaded with a calcium responsive fluorescent probe, to filtration through 5 and 3 microm pores. Calcium entered 28% of normal red cells at the moment of deformation when 3 microm filtered, a finding consistent with the transient activation of a mechanosensitive channel. Red cells containing hemoglobin AC and AS had enhanced calcium responses to filtration. An increased influx of calcium in hemoglobin disorders might play a role in providing protection against Falciparum malaria. PMID- 15121091 TI - Excess haemoglobin digestion by malaria parasites: a strategy to prevent premature host cell lysis. AB - To understand the osmotic stability of a Plasmodium falciparum-infected red blood cell, whose membrane permeability becomes highly increased during parasite growth, we developed an integrated mathematical model of the homeostasis of an infected red cell. The model encoded the known time courses of red cell membrane permeabilisation and of haemoglobin digestion, as well as alternative options for parasite volume growth. Model simulations revealed that excess haemoglobin digestion, by reducing the colloid-osmotic pressure within the host red cell, is essential to preserve the osmotic stability of the infected cell for the duration of the parasite asexual cycle. We present here experimental tests of the model predictions and discuss the available evidence in the context of the interpretations provided by the model. PMID- 15121092 TI - Use of luminescence resonance energy transfer to measure distances in the AE1 anion exchange protein dimer. AB - To understand how red blood cell and other proteins carry out their functions, it is necessary not only to have high-resolution crystal structures, but also to have methods that can measure changes in position of parts of the protein on the scale of Angstroms. The method of luminescence resonance energy transfer (LRET) has considerable advantages for this purpose, particularly for proteins, such as the AE1 anion exchange protein in the red cell, that are homodimers. We have applied this method, using a terbium maleimide chelate (TbM) as donor and fluorescein maleimide (FM) as acceptor, to measure the distance between the C201 residues in adjacent dimerized cytoplasmic domains of AE1 (cdAE1). The distance measured by LRET (40.8 A) corresponds closely with that calculated from the crystal structure of the cdAE1, indicating that the method can provide useful information for testing hypotheses concerning motions in this and other blood cell proteins. PMID- 15121093 TI - Anionic channels in malaria-infected human red blood cells. AB - As the intraerythrocytic stage of the human malarial parasite, Plasmodium falciparum, matures, the plasma membrane of the host red blood cell (RBC) becomes increasingly permeable to a variety of physiologically relevant solutes via the induction of new permeation pathways (NPPs) (H. Ginsburg, Novartis Foundation Symposium 226, 99-108,1999; K. Kirk, Physiol. Rev. 81, 495-537, 2001). Although permeable to cationic and electroneutral solutes, transport studies have shown that the NPPs exhibit the general properties of anion channels and recent electrophysiological studies, using the patch-clamp technique, have demonstrated that anion channels are activated in the plasma membrane of the RBC following infection (S.A. Desai et al., Nature 406, 1001-1005, 2000; S.M. Huber et al., EMBO J. 21, 22-30,2002; S. Egee et al., J. Physiol. 542, 795-801, 2002). In this paper, we review the features of the anionic channels that we have observed in both uninfected and malaria-infected human RBCs, the data that suggest that the NPPs are endogenous to the RBC membrane, and present new evidence, which suggests that the mechanism of induction of the NPPs, used by the parasite, involves phosphorylation steps. PMID- 15121094 TI - Slow transitions between two conformational states of band 3 (AE1) modulate divalent anion transport and DBDS binding to a second site on band 3 which is activated by lowering the pH (pK approximately 5.0). AB - Evidence is emerging which indicates that the anion transport activity of band 3 may be regulated. I review the molecular basis for regulation of the anion transport function of band 3 in terms of evidence showing that divalent anion transport involves a slow "hysteretic" transition between two functional states, mediated by interactions between subunits within band 3 oligomers. In addition, I briefly describe recent work from my laboratory where we have discovered a novel manifestation of slow conformational changes in band 3. This involves 4,4' dibenzamido-2,2'-stilbenedisulfonate (DBDS) binding to a second, proton-activated site distinct from the primary stilbenedisulfonate site. This site is exposed on the outer aspect of band 3 when the pH is lowered (pK approximately 5.0). This is the same pK as the protonation site on band 3 involved in divalent anion-proton co-transport (APCT) [J. Gen. Physiol. 79 (1982) 87]. Significantly, we have found that DBDS binding to this proton-activated site has unusually slow kinetics, and increasing DBDS concentration causes a decrease in the apparent pseudo-first order rate constant. These results suggest that a slow conformational pre equilibrium is the rate limiting step in DBDS binding to the proton-activated site on band 3 observed at low pH. Our results support an allosteric two-state model for regulation of divalent anion transport by band 3 oligomers involving a slow conformational transition and interactions between subunits [Biochemistry 31 (1992) 7301]. PMID- 15121095 TI - The disruption of the third extracellular loop of the red cell anion exchanger AE1 does not affect electroneutral Cl-/HCO3- exchange activity. AB - The red cell anion exchanger (band 3; AE1) is a multispanning membrane protein that traverses the bilayer up to 14 times and mediates the stilbene-disulfonate sensitive, electroneutral exchange of chloride and bicarbonate. Previous studies showed that the integrity of the third extracellular loop (EC3) of the protein was not essential for stilbene-disulfonate-sensitive chloride uptake. Here we demonstrate that the chloride uptake mediated by assemblies separated at EC3 represents the physiological electroneutral Cl(-)/HCO(3)(-) activity associated with intact AE1 protein. This provides further evidence that the 1:5 and 6:14 regions of the protein form discrete folding domains and confirms that the third extracellular loop does not play a pivotal role in AE1 transport function. PMID- 15121096 TI - Pharmacology of the human red cell voltage-dependent cation channel; Part I. Activation by clotrimazole and analogues. AB - The activation and pharmacological modulation of the nonselective voltage dependent cation (NSVDC) channel from human erythrocytes were studied. Basic channel activation was achieved by suspending red cells in a low Cl(-) Ringer (2 mM), where a positive membrane potential (V(m) = E(Cl)) immediately developed. Voltage- and time-dependent activation of the NSVDC channel occurred, reaching a cation conductance (g+) of 1.5-2.0 microS cm(-2). In the presence of the classical Gardos channel blocker clotrimazole (0-50 microM), activation occurred faster, and g+ saturated dose-dependently (EC50 = 14 microM) at a value of about 4 microS cm(-2). The clotrimazole analogues TRAM-34, econazole, and miconazole also stimulated the channel, whereas the chemically more distant Gardos channel inhibitors nitrendipine and cetiedil had no effects. Although the potency for modulation of the NSVDC channel is much lower than the IC50 value for Gardos channel inhibition, clotrimazole (and its analogues) constitutes the first chemical class of positive modulators of the NSVDC channel. This may be an important pharmacological "fingerprint" in the identification of the cloned equivalent of the erythrocyte channel. PMID- 15121097 TI - Cell shrinkage activates Na+/H+ exchange in dog red cells by relieving inhibition of exchange by Na+ in isotonic medium. AB - Na+/H+ exchangers (NHE) are widely distributed transporters responsible for regulation of cell volume and pH. In isotonic medium, NHE is often low or negligible, and is strongly activated by osmotic cell shrinkage. It is reported here that extracellular Na+ inhibits NHE in isotonic medium, and cell shrinkage stimulates NHE by relieving this inhibition. There is more than one inhibitory Na+ site on each transporter. Shrinkage activates NHE by decreasing the apparent affinity for Na+ at the inhibitory sites. Shrinkage has no effect on the apparent affinity for Na+ at the substrate sites for activation of NHE. PMID- 15121099 TI - Conventional transport assays underestimate sugar transport rates in human red cells. AB - The time course of protein-mediated 3-O-methylglucose uptake by human red cells and by red cell ghosts containing or lacking 4 mM MgATP was measured at ice temperature and sub-saturating sugar levels by conventional sampling procedures and at 20 degrees C by use of a quench-flow apparatus. The temporal resolution of the quench-flow apparatus (as fast as 5-ms sample times) was confirmed by analysis of alkaline hydrolysis of dinitrophenolacetate. Red cell sugar uptake at 4 degrees C is consistent with two processes [fast (tau = 120 s) and slow (tau = 1100 s)] that occur in series. Intracellular ATP increases the size and the rate of equilibration of the fast compartment and slows the rate of filling of the slow compartment. Red cell ghost volume and protein content are unaffected by lysis/resealing in the presence of ATP. Uptake at 20 degrees C is also consistent with two processes [fast (tau = 10 ms) and slow (tau = 15 s)] that occur in series. ATP increases the size of both compartments and the rate of filling of the small compartment at 20 degrees C. Preliminary estimates indicate that the sugar uptake capacity of human red cells at 20 degrees C is underestimated by as much as 8-fold by measuring sugar uptake over 2 s vs. 26 ms. We discuss the implications of multiphasic sugar uptake in the context of models for protein mediated sugar transport. PMID- 15121098 TI - Bretylium, an organic quaternary amine, inhibits the Na,K-ATPase by binding to the extracellular K-site. AB - The quaternary amine, bretylium, is a class III antiarrhythmic drug used to treat ventricular tachycardia and fibrillation. The primary mode of action for bretylium is thought to be inhibition of voltage-gated K(+) channels. While the Na,K-ATPase has been the pharmacological target of cardiac glycosides for over a century, recent evidence has shown that bretylium may also inhibit the Na pump. Our experimental findings support and extend these previous reports and provide definitive evidence supporting the previous suggestion that bretylium and K compete for the Na pump. We find that bretylium inhibits the Na pump in a dose dependent manner in both Na,K-ATPase (IC(50) 4.5 mM) and Rb flux experiments (IC(50) 3.5 mM). Furthermore, we show that bretylium and Rb(+) competes for an extracellular site by measuring ouabain-sensitive (86)Rb flux in intact human red blood cells; that is, there is an apparent increase in K(m) for Rb(+) in the presence of 5 mM bretylium, while V(max) remains unchanged. We also determined that unlike K(+), bretylium does not facilitate the hydrolysis of E2-P. However, it stabilizes this conformation by reducing the ability of K(+) to facilitate dephosphorylation. Finally, we show that bretylium, like K(+), reduces [(3)H]ouabain binding to the Na pump. Taken together, these data are consistent with bretylium binding to the extracellular facing cation site within the E2-P state of the enzyme. Moreover, these findings suggest that bretylium may serve as an effective tool for freezing the pump in an extracellularly cation-bound phosphorylated intermediate, which will aid in future structural analyses. PMID- 15121100 TI - Antibodies to betaISigma2 spectrin identify in-homogeneities in the erythrocyte membrane skeleton. AB - The cortical cytoskeleton of the mammalian red cell, composed of spectrin, actin, protein 4.1, adducin, and protein 4.9, is generally regarded as a homogeneous structure that maintains the integrity of the membrane and the lateral disposition of integral membrane proteins. The major component of this structure is a hetero-oligomer of alphaI and betaISigma1 spectrin. In other tissues, most notably muscle and brain, a transcript of the betaI spectrin gene is generated by alternative exon utilization, yielding a protein that has the COOH-terminal 19 residues of betaISigma1 spectrin replaced by 210 novel residues to generate betaISigma2 spectrin. This new transcript contains a pleckstrin homology (PH) domain and may even exist under some conditions in a homopolymeric form. Using antibodies specific for the COOH-terminal domains of either betaISigma1 or betaISigma2 spectrin, we find that contrary to previous understandings, mature human erythrocytes contain a subpopulation of spectrin that is immunoreactive with antibodies to the betaISigma2 isoform, and that this spectrin is distributed into distinct plasma membrane patches. These results suggest that the native mammalian erythrocyte membrane skeleton, rather than being homogeneous, contains discrete submicron-scale microdomains that differ in both their composition and dispersion across the cell surface. The precise nature and role of these putative microdomains is under active investigation. PMID- 15121101 TI - Eukaryotic and prokaryotic stomatins: the proteolytic link. AB - The 32kD membrane protein stomatin was first studied because it is deficient from the red cell membrane in two forms of the class of haemolytic anaemias known as "hereditary stomatocytosis." The hallmark of these conditions is a plasma membrane leak to the monovalent cations Na+ and K+: the protein is missing only in the most severely leaky of these conditions. No mutation has ever been found in the stomatin gene in these conditions. Stomatin-like proteins have been identified in all three domains of biology, yet their function remains enigmatic. Although the murine knock-out is without phenotype, we have identified a family showing a splicing defect in the stomatin mRNA, in which affected children showed a catastrophic multisystem disease not inconsistent with the now-known wide tissue distribution of stomatin. We report here a study of strongly homologous stomatin-like genes in prokaryotes, which reveals a close connection with a never studied gene erroneously known as "nfed." This gene codes for a hydrophobic protein with a probable serine protease motif. It is possible that these stomatin like genes and those which are known as"nfed" form an operon, suggesting that the two protein products are aimed at a common function. The corollary is that stomatin could be a partner protein for a membrane-bound proteolytic process, in both prokaryotes and in eukaryotes generally: this idea is consistent with experimental evidence. PMID- 15121102 TI - Emerging role of nitrite in human biology. AB - Nitric oxide (NO) plays a fundamental role in maintaining normal vascular function. NO is produced by endothelial cells and diffuses both into smooth muscle causing vasodilation and into the vessel lumen where the majority of this highly potent gas is rapidly inactivated by dioxygenation reaction with oxyhemoglobin to form nitrate. Diffusional barriers for NO around the erythrocyte and along the endothelium in laminar flowing blood reduce the inactivation reaction of NO by hemoglobin, allowing sufficient NO to escape for vasodilation and also to react in plasma and tissues to form nitrite anions (NO(2)(-)) and NO modified peptides and proteins (RX-NO). Several recent studies have highlighted the importance of the nitrite anion in human biology. These studies have shown that measurement of plasma nitrite is a sensitive index of constitutive NO synthesis, suggesting that it may be useful as a marker of endothelial function. Additionally, recent evidence suggests that nitrite represents a circulating storage pool of NO and may selectively donate NO to hypoxic vascular beds. The conversion of nitrite to NO requires a reaction with a deoxygenated heme protein, suggesting a novel function of hemoglobin as a deoxygenation-dependent nitrite reductase. This review focuses on the role of nitrite as a circulating NO donor, its potential as an index of NO synthase (NOS) activity and endothelial function, and discusses potential diagnostic and therapeutic applications. PMID- 15121103 TI - Phosphatidylserine binding sites in red cell spectrin. AB - Spectrin has been shown to interact with phosphatidylserine (PS), however, the precise binding sites for PS in spectrin have not been defined. In the present study, we have identified specific PS binding sites in spectrin using recombinant spectrin fragments encompassing the entire sequences of both spectrin chains. We show that sites of high affinity are located within eight of the 38 triple helical structural repeats which make up the bulk of both chains: these are: alpha8 and alpha9-10, and beta2, beta3, beta4, beta12, beta13 and beta14, and PS affinity was also found in the non-homologous N-terminal domain of the beta chain. It is noteworthy that the PS-binding sites in beta-spectrin are clustered in close proximity to the sites of attachment both of ankyrin and of 4.1R, the proteins engaged in attachment of spectrin to the membrane. We conjecture that direct interaction of spectrin with PS in the membrane complements modulates its interactions with the proteins, and that (considering also the known affinity of 4.1R for PS) the formation of PS-rich lipid domains, which have been observed in the red cell membrane, may be a result. PMID- 15121104 TI - Endovascular treatment of mycotic aneurysms of the thoracic and abdominal aorta: the need for level I evidence. PMID- 15121105 TI - Current therapeutic options for treating primary hyperhidrosis. AB - Severe hyperhidrosis can cause extreme embarrassment that may lead to social and professional isolation. Therapeutic strategies to hyperhidrosis should employ the least invasive treatment that provides effective symptom control. The treatment options available for control of hyperhidrosis, non-surgical or surgical, differ in their invasiveness and efficacy. Mechanisms of action of antiperspirants, iontophoresis, cholinergic inhibitor drugs, botulinum toxin, and surgical sympathectomy are reviewed. There is little published evidence in the form of comparative randomised trials to support the use of one treatment over another. However, authors have tended to recommend those therapies that are available to their speciality. Specific therapies should be tailored to the patient's symptoms to gain maximum symptomatic improvement with minimum invasiveness and side effects. To achieve this, the full range of treatment options should be available to, or accessible by the consulting doctor in order for the patient to have a meaningful choice. PMID- 15121106 TI - Current diagnosis and management of blunt internal carotid artery injury. AB - BACKGROUND: Blunt carotid artery injury (BCI) is a rare but potentially devastating injury. When undiagnosed it can result in severe disability or death. METHODS: A Medline-based literature search was performed using key words 'blunt carotid injury' and cross-referenced with further original papers obtained from the references from this search. RESULTS AND CONCLUSIONS: The incidence of BCI is very low. However, given the serious consequences of a missed injury, recent efforts have focussed on targeted screening for this injury in trauma patients. Conventional angiography remains the investigation of choice but may be superceded in the future by non-invasive methods such as magnetic resonance angiography or CT angiography. Operative intervention is rarely required and anti coagulation remains the treatment of choice where dissection or pseudoaneurysm is diagnosed. The role of anti-platelet therapy is currently being investigated. Endovascular management using stents has been described but medium to long term results are not yet available. PMID- 15121107 TI - 11-year experience with anatomical and extra-anatomical repair of mycotic aortic aneurysms. AB - BACKGROUND: We have reviewed our management, of both ruptured and non-ruptured, abdominal and thoraco-abdominal mycotic aneurysms in order to determine the safety and efficacy of in situ and extra-anatomical prosthetic repairs. METHODS: Data regarding presenting symptoms, investigations, operative techniques and outcome, were collected on patients treated at a singe centre over 11 years. RESULTS: There were 11 men and four women, with a median age of 70 years (range, 24-79). All but one patient were symptomatic and six had a contained leak on admission. In six patients no organisms were identified in either blood or tissue cultures. Pre-operative CT identified; four infra-renal, four juxta-renal, three (Crawford thoraco-abdominal) type IV, three type III and one type II, aortic aneurysms. Thirteen were repaired with in situ prostheses and two required axillo femoral prosthetic grafts. There were four early deaths. All surviving patients have been followed-up for a median duration of 38 months (range 1/2-112 months). There were two late deaths at 3 months (juxta-renal) and at 2 years (type III), the latter relating to graft infection. CONCLUSIONS: In the absence of uncontrolled sepsis, repair of mycotic aortic aneurysms using prosthetic grafts can achieve durable results. PMID- 15121108 TI - Cryopreserved arterial allografts in the treatment of prosthetic graft infections. AB - AIM: The purpose of this study was to evaluate the effectiveness of cryopreserved arterial allografts in the management of prosthetic graft infection. MATERIAL AND METHODS: Over a 5-year period 45 patients with infection of prosthetic vascular grafts were treated. There were 39 intra-abdominal infected grafts (group I) and six extra-abdominal infected grafts (group II). Treatment consisted of total graft removal and in situ or extra-anatomic implantation of cryopreserved arterial allografts. Six patients were operated on as an emergency. Four patients presented with aorto-enteric fistula. Follow-up ranged from 30 to 78 months. RESULTS: There were six in-hospital deaths and two additional patient deaths during follow-up, yielding an overall mortality rate of 18%. Six patients died due to complications directly related to infection or insertion of an allograft. Combined short and long-term mortality rate was much higher in patients operated on as an emergency (67%) compared to elective cases (11%). Patients with aorto enteric fistula had the highest mortality rate (75%). Primary and secondary 3 year allograft patency rates for group I were 84 and 94%, respectively and for group II were 60 and 80%, respectively. CONCLUSIONS: Aortic allografts are useful in the treatment of infection of major vascular prosthetic grafts, except for patients with aorto-enteric fistula. Patients with infection of the prosthetic graft should be promptly assessed for graft removal, since results of elective surgery are much better than results of emergency procedures. PMID- 15121109 TI - Cryopreserved arterial allografts for in situ reconstruction of infected arterial vessels. AB - OBJECTIVE: To review our experience of using cryopreserved allografts for in situ reconstruction in the presence of infection involving the aorta, iliac or femoral arteries. DESIGN: Retrospective clinical study. METHODS: From 3/2000 to 8/2003 all patients with mycotic aneurysms or secondary infection following earlier prosthetic replacement were treated with cryopreserved human allografts. Forty two patients, 39 (93%) with a prosthetic graft infection and 3 (7%) with a mycotic aneurysm of the abdominal aorta were treated. Six (14%) had aorto-enteric fistulas, 5 (12%) had ruptured aneurysms, and 2 also had vertebral destruction. The median follow-up time was 20 months (range 1-42 months). RESULTS: Thirty-day mortality was 14%. Three patients died due to multi-organ failure, two patients died from hypovolaemic shock due to allograft rupture and one from rupture of the native aorta. The overall mortality was 24% (four additional patients). Graft patency was 100% at 30 days and 97% at follow up in the survivors. The mean actuarial survival time was 32 months (95% CI=27-37 months). CONCLUSIONS: Cryopreserved allografts for the in situ reconstruction of infected arteries or grafts have acceptable intermediate results. PMID- 15121110 TI - RNAIII-inhibiting peptide and/or nisin inhibit experimental vascular graft infection with methicillin-susceptible and methicillin-resistant Staphylococcus epidermidis. AB - OBJECTIVE: To investigate the efficacy of RNAIII-inhibiting peptide (RIP) and nisin as prophylactic agents in a rat model of vascular graft infection. DESIGN: Prospective, randomized, controlled animal study. MATERIALS: Two hundred and twenty adult male Wistar rats. Staphylococcus epidermidis ATCC 12228 and one clinical isolate of methicillin-resistant S. epidermidis. Drugs: RIP, nisin and rifampin. METHODS: Graft infections were established in the dorsal subcutaneous tissue by implantation of 1 cm(2) sterile Dacron grafts, followed by topical bacterial inoculation: grafts were retrieved at 7 days. The study included a control group (without inoculation) and two series composed of five groups for each staphylococcal strain: one contaminated group that did not receive any antibiotic prophylaxis, three contaminated groups that received grafts soaked with 10 mg/l RIP, 10 mg/l nisin, 10 mg/l rifampin, or RIP+nisin. The main outcome measure was the extent of bacterial at graft harvest. RESULTS: The bacterial counts for methicillin-resistant S. epidermidis on explanted grafts were 6.1+/ 2.8x10(2), 7.8+/-3.0x10(3) and 5.5+/-2.9x10(4) for RIP, nisin and rifampin, respectively. RIP and nisin used in combination reduced the bacterial count to <10. The results for S. epidermidis were similar. CONCLUSIONS: RIP and nisin could be used in combination to coat medical devices to prevent drug resistant S. epidermidis infections. PMID- 15121111 TI - Cellular engineering of conduits for coronary and lower limb bypass surgery: role of cell attachment peptides and pre-conditioning in optimising smooth muscle cells (SMC) adherence to compliant poly(carbonate-urea)urethane (MyoLink) scaffolds. AB - OBJECTIVE: We are developing a hybrid arterial bypass graft of compliant poly(carbonate-urea)urethane (MyoLink), endothelial and smooth muscle cells (SMCs). To enhance adhesion of SMCs we assessed various attachment factors and the effect of pre-conditioning on cell retention. METHODS: MyoLink segments were coated with either RGD, superfibronectin, fibronectin, fibronectin-like engineered polymer protein (FEPP), FEPP plus or type 1 collagen overnight. (111)Indium-radiolabelled SMCs were placed onto MyoLink segments for 48 h before being aspirated, then lavaged off. All grafts, aspirates and lavages were counted in a gamma counter. SMC viability on the MyoLink segments was also assessed for viability using the Alamar blue redox assay. Separately, MyoLink grafts lined with radiolabelled SMCs were divided into a pre-conditioned group, exposed to subarterial pulsatile flow whilst another group were held in static culture. After 1-week, grafts were exposed to arterial pulsatile flow whilst radioactivity was assessed using a gamma camera. RESULTS: Only FEPP plus significantly enhanced SMC attachment: mean of 32+/-6% cell attachment compared to 21+/-5% for uncoated control. Cell viability was enhanced by all attachment factors except fibronectin. Pre-conditioning was shown to significantly enhance the retention of SMCs onto the MyoLink once exposed to pulsatile arterial flow: the final attachment was 57+/-7% for the static and 76+/-7% for the pre-conditioned group. CONCLUSIONS: FEPP plus enhances SMC attachment to MyoLink. We believe this is because of its repeating sequences of RGD and its positive charge. Pre conditioning enhances the retention of SMCs to MyoLink once exposed to pulsatile arterial flow. PMID- 15121112 TI - Interposition vein cuff and intimal hyperplasia: an experimental study. AB - OBJECTIVE: There is some evidence to suggest that prosthetic distal bypass graft patency can be improved, and the risk of intimal hyperplasia diminished, by interposing a distal vein cuff. We studied intimal remodeling in an end-to-side distal prosthetic anastomosis constructed with and without a vein cuff. METHODS: Twenty-four prosthetic bypasses were constructed with (N=12) or without (N=12) a distal vein cuff in 12 pigs. At 10 weeks, the 20 anastomoses and adjacent arteries from the surviving 10 pigs were studied by histology, immunohistochemistry and morphometry. RESULTS: Intimal hyperplasia was significantly less on all zones of the arterial floor and all suture zone of arteries anastomosed with a vein cuff than within arteries anastomosed without a vein cuff (0.11 versus 0.34; p=0.001 and 0.35 versus 1.19; p=0.0001, respectively). Intimal hyperplasia was also more prominent within the vein cuff than within the recipient artery, with or without a vein cuff (1.35 versus 0.38; p=0.0001). CONCLUSION: An interposition vein cuff at the distal anastomosis between a prosthesis and an artery alters the distribution of intimal hyperplasia. By acting as an expansion chamber where intimal hyperplasia can develop harmlessly, the vein cuff may protect the arterial anastomosis from stenosis. PMID- 15121113 TI - Peripheral arterial disease: public and patient awareness in The Netherlands. AB - OBJECTIVE: To determine critical issues for future awareness programmes on peripheral arterial disease (PAD). DESIGN: National Dutch survey. MATERIALS AND METHODS: A representative sample of 1294 members of the general population, and 281 patients with PAD from the Capi@home database were administered a questionnaire concerning awareness of PAD. RESULTS: The response rate was 81% for the general population and 78% for patients with PAD. The familiarity with PAD terminology and symptoms amongst the general population was low. Few patients (20%) were aware that PAD was a disease of arteries. Amongst both the general population and the patient populations, PAD risk factors identification was low: hypertension (4% versus 0%); hypercholesterolaemia (9% versus 12%), diabetes (2% versus 8%), and smoking (27% versus 52%). Knowledge was moderate in both populations about treatment with exercise, but low for smoking cessation. The general population was unaware of the central role of general practitioners in the treatment of PAD. CONCLUSIONS: The awareness of symptoms, risk factors, and treatment options for PAD is low. Both population and patients needed only minimal information to relate PAD to other atherosclerotic diseases. Based on the results of this survey the Dutch Platform of Peripheral Arterial Disease together with the Dutch Heart Foundation are initiating the first awareness campaign on atherosclerosis. PMID- 15121114 TI - Toe blood pressure measurements in patients suspected of leg ischaemia: a new laser Doppler device compared with photoplethysmography. AB - OBJECTIVES: To study the usefulness of a new laser Doppler (LD) device as compared with photoplethysmography (PPG) to measure toe systolic blood pressures (TBP) in patients suspected of arterial insufficiency of the legs. DESIGN: Prospective comparative clinical study. MATERIALS AND METHODS: Seventy patients (121 legs) referred to our vascular laboratory for TBP measurements were investigated using a standard PPG and a new LD instrument, incorporating a cuff inflator and probe heater. RESULTS: Mean toe pressures with the PPG, unheated LD, and heated LD (37 degrees C) were 68.1, 67.2, and 75.5 mmHg, respectively. TBP with heated LD was systematically higher than with PPG (7.7 mmHg; p<0.001). Intra class correlations between PPG vs. unheated LD and PPG vs. heated LD were good: 0.91 and 0.93, respectively. However, a considerable variation existed between PPG and LD (95% confidence interval: 45 mmHg), which were unrelated to the value of the TBP. Kappa values of agreement between PPG and unheated and heated LD to detect critical ischaemia (TBP below 30 mmHg) were good (kappa=0.77 and 0.63, respectively). CONCLUSIONS: The new LD device appears a good alternative to standard PPG measurements to assess TBP, despite a rather wide variation. PMID- 15121115 TI - Limb-salvage by femoro-distal bypass and free muscle flap transfer. AB - OBJECTIVES: To evaluate the feasibility and long-term outcome of distal arterial reconstruction combined with free muscle flap transfer for patients who would otherwise have undergone major amputation. METHODS: Between 1996 and 2001, 27 reconstructions using autologous vein were performed in 25 patients. Seventeen of these patients had diabetes mellitus. Gracilis, rectus abdominis and latissimus dorsi muscles were used as free flaps, covered with split-thickness skin grafts. RESULTS: Eighty-five percent of patients had a patent graft and viable muscle flap after 1-month. Mean follow-up was 51 months (4-72 months). At the time of follow-up 77% of reconstructions were patent and 70% of patients regained full functional capacity of their lower extremities. CONCLUSION: Limb-salvage by distal arterial reconstruction and free muscle flap transfer, is feasible with low mortality and morbidity and provides excellent long-term results with regard to graft patency and functional status. PMID- 15121116 TI - Prediction of cerebral ischemic tolerance during carotid cross-clamping by angiographic criteria. AB - OBJECTIVE: To evaluate the prognostic value of angiographic criteria and internal carotid artery (ICA) stump pressure for shunt placement in carotid surgery under local anaesthesia. DESIGN: Prospective clinical trial at a university hospital. MATERIALS AND METHODS: In 120 patients a cerebral angiography was initiated before undergoing carotid surgery. Seventy-five percent of the patients were neurologically asymptomatic, 13% had transient ischaemic attacks and 12% suffered from previous strokes. The operation was exclusively performed under local anaesthesia and prior to cross-clamping the ICA stump-pressure was measured. A shunt was inserted only if hemispheric symptoms or unconsciousness occurred independent of the angiographic findings or stump pressure. RESULTS: In 23% (27/120) a shunt became necessary and significantly (p<0.001) more often when there was a cross-flow towards the contralateral hemisphere (12/20=60%) or if the contralateral ICA was occluded (9/13=69%). The sensitivity for not needing a shunt in case of cross-flow towards the side of operation was 91% (52/57) whereas the specificity was 35% (22/63). ICA stump-pressure was significantly reduced in patients requiring a shunt (31 mmHg) compared to those not needing a shunt (53 mmHg) (p<0.001), but no definitive threshold value was found determining the need for shunting. Intraoperatively, no persistent neurological complication developed. CONCLUSIONS: Angiographic cross-flow was a good, but not perfect predictor for the need of an intraoperative shunt. PMID- 15121117 TI - Near infrared spectroscopy monitoring during carotid endarterectomy: which threshold value is critical? AB - OBJECTIVES: Retrospectively to verify which decreasing percentage in regional oxygen saturation (rSO(2)) identified patients with good collateralisation during carotid artery cross clamp. MATERIALS AND METHODS: During 594 endarterectomies under general anaesthesia the decreasing percentage from preclamp value to value detected in the first 2 min after clamping the CCA and/or ICA was calculated in real time. No temporary shunt was placed in any case. ROC analysis was performed to determine the optimal cut-off for rSO(2) decrease to identify the occurrence of neurological complications. RESULTS: A cut-off of 11.7% was identified as optimal. Sensitivity and specificity were 75% (95% CI 71-78) and 77% (95% CI 74 80), respectively. The cut-off of 20% had a lower sensitivity (30%) and a higher specificity (98%) to identify patients with complications, with positive and negative predictive value of 37 and 98%, respectively. CONCLUSIONS: The study suggest that a relative decrease in rSO(2) of <20% from preclamp to early cross clamp value has a high negative predictive value, i.e. if rSO(2) does non decrease more than 20%, ischemia by hypoperfusion is unlikely and a shunt should not be necessary. Moreover, a relative decrease >20% may not always indicate intraoperative neurological complications. PMID- 15121118 TI - Carotid endarterectomy relieves pulsatile tinnitus associated with severe ipsilateral carotid stenosis. AB - OBJECTIVES: Pulsatile tinnitus is a rare and often disabling condition. Pulsatile tinnitus sometimes occurs in patients with severe atherosclerotic carotid stenosis. It is uncertain whether carotid endarterectomy (CEA) relieves pulsatile tinnitus in patients with severe carotid stenosis. DESIGN, MATERIALS AND METHODS: This is a retrospective study of 14 patients with pulsatile tinnitus who underwent CEA. Demographic and clinical features and pre-operative duplex results were recorded. Operative results in this group were assessed. RESULTS: CEA relieved symptoms of pulsatile tinnitus in 10 out of 14 cases (70%). Of 10 patients that had lateralisable tinnitus and ipsilateral surgery, 9 (90%) reported symptomatic improvement. CONCLUSIONS: CEA is effective in improving pulsatile tinnitus in patients with unilateral symptoms and severe ipsilateral carotid stenosis. PMID- 15121119 TI - Patient satisfaction for carotid endarterectomy performed under local anaesthesia. AB - OBJECTIVES: To develop, validate and use a procedure specific questionnaire to evaluate patient experience and satisfaction following carotid endarterectomy (CEA) under either general (GA) or local anaesthesia (LA). METHODS: Twenty post CEA patients were interviewed. Data were content analysed and recurrent themes used to generate the specific carotid endarterectomy experience questionnaire (CEA-EQ). The CEA-EQ consists of 15 pre-op and 13 postoperative questions. Validity was established by correlation with the FRS Patient Satisfaction with Surgical Services (SSSQ) and State form of the State Trait Anxiety Inventory (STAI-S) in 69 patients (35 LA, 34 GA). Subsequently 88 patients randomised to GA CEA and 88 to LA CEA received the CEA-EQ. A local anaesthetic intraoperative experience questionnaire (LA-EQ) was also developed and given to LA patients only. RESULTS: Validity was confirmed through significant correlations with the STAI-S (r=0.67, p<0.001) and the SSSQ (r=0.44, p<0.001). In the randomised prospective study response rates were greater than 90%. Overall experience and satisfaction with CEA was high. There was no statistically significant difference in anxiety, satisfaction or overall experience between anaesthetic techniques. LA CEA was associated with a significantly better perception of recovery. The majority of LA patients found the procedure acceptable. CONCLUSIONS: The CEA-EQ is a valid tool to assess qualitative aspects of CEA patient care. Overall satisfaction and experience with CEA is good and not related to anaesthetic technique. LA CEA is not associated with any increased anxiety, is tolerated by the majority of patients and is associated with a better perception of recovery. PMID- 15121120 TI - Intermediate outcome and risk factor assessment of bovine vascular heterografts used as AV-fistulas for hemodialysis access. AB - OBJECTIVES: This observational study was set up to prospectively follow all bovine heterograft (ProCol) fistulas implanted for hemodialysis access between 1998 and 2002. METHODS: ProCol was implanted if autogenous vein was not available or if patients presented with a history of failed, infected or otherwise complicated ePTFE grafts and/or on immunosuppressive therapy. Fistula patency was the primary outcome; secondary outcomes were clinical events and the rate of access revisions. RESULTS: Sixty-two ProCol grafts were implanted in 56 patients. The mean primary (PP) and secondary patency (SP) was 334 (SEM 57) and 528 (SEM 59) days, respectively. Coronary heart disease was associated with a significantly better SP (OR 0.2, 95% CI 0.1-0.9) whilst diabetes mellitus was associated with a significantly worse SP (OR 0.2, 95% CI 0.1-0.9). Reinterventions were performed at a mean rate of 1.23 (SEM 0.17) per fistula. The relative risk of access revision was significantly higher in patients with diabetes mellitus (OR 9.2, 95% CI 2.3-37.2). CONCLUSIONS: ProCol grafts, used for AV-fistulas, demonstrate acceptable patency rates in high-risk haemodialysis patients. Diabetes mellitus jeopardizes the patency of these fistulas and is associated with a high revision rate. PMID- 15121121 TI - Treatment of venous leg ulcers with Dermagraft. AB - BACKGROUND: A number of different treatment approaches have been recommended for the treatment of venous ulceration, including local ulcer treatment, compression and drug therapy. Recent advances in tissue engineering have resulted in living tissues being developed for cutaneous wound repair and skin replacement. The aim of this pilot study was to compare the rate of healing of venous ulcers in patients treated with Dermagraft (a human fibroblast-derived dermal replacement) and compression therapy or compression therapy alone. METHODS: A total of 18 patients with venous ulceration of the leg were recruited into the pilot study. Ten patients were treated with Dermagraft and compression therapy, and eight patients were treated with compression therapy alone. Healing was assessed by ulcer tracing and computerised planimetry. Skin perfusion was measured by laser Doppler. RESULTS: Five (50%) of the patients treated with Dermagraft and one (12.5%) control patient had healed by the end of the 12-week study period (NS). The total ulcer area rate of healing and linear rate of healing was significantly improved in patients treated with Dermagraft (P=0.001 and P=0.006, respectively, Mann-Whitney U-test). The number of capillaries increased in both the treatment and control group. Peri-ulcer skin perfusion increased by 20% in patients treated with Dermagraft, compared with 4.9% in the control group. CONCLUSION: The data from this small pilot study suggests that Dermagraft is associated with improved healing of venous ulceration. Following this pilot study, further clinical studies are needed to confirm the validity of these results in 'hard to heal' venous leg ulcers. PMID- 15121122 TI - Endovascular treatment of a MRSA infected left external iliac artery pseudoaneurysm. PMID- 15121123 TI - A new type of magnification system in vascular surgery--an evaluation study. PMID- 15121124 TI - Primary aorto-duodenal fistula following Staphylococcal septicaemia. PMID- 15121126 TI - Frequency of anticardiolipin, antinuclear and anti beta2GP1 antibodies is not increased in unselected epileptic patients: a case-control study. AB - In order to determine whether auto antibodies were restricted to some subtypes of epilepsy, we included 81 unselected epileptic patients and 81 controls, studied clinical data, EEG, neuroimaging, measured antinuclear (ANA), anticardiolipin (aCL) and beta2GP1 antibodies. RESULTS: Epilepsy was active in 74 patients, generalised in 78 and partial in 9. Auto antibodies were positive at the same frequency and the same level among patients and controls (ANA+ 17% vs. 11%; aCL+ 4% vs. 7%; beta2GP1 antibodies+ 5% vs. 6%). There was no increased frequency of auto antibodies among subgroups of epileptic patients except ANA which were more frequent when patients had more than 10 seizures per year. CONCLUSIONS: Positivity of ANA, aCL and beta2GP1 antibodies is not increased in all types of epilepsy and further studies are needed to determine exactly which kind of seizure is immune-mediated. PMID- 15121127 TI - The influence of gender on the aggravation of absence seizures by carbamazepine in the low-dose pentylenetetrazol rat model. AB - OBJECTIVES: To determine whether carbamazepine (CBZ) aggravates absence seizures in the low-dose pentylenetetrazol (PTZ) rat model in both male and female animals, and investigate for gender differences. METHODS: Inbred Sprague-Dawley rats were implanted with EEG electrodes. Seven days later PTZ (20 mg/kg, i.p.) was administered following pre-treatment with vehicle or CBZ (20 mg/kg, i.p.) and the occurrence of spike-and-wave discharges (SWDs) on the EEG quantified. RESULTS: The cumulative SWD for 90-minute post-PTZ was higher in the CBZ versus vehicle pre-treatment arm for both female (mean 110 seconds vs. 62 seconds; P = 0.03) and male (mean 89 seconds vs. 60 seconds; P = 0.03) rats. The increase in SWD duration in the CBZ arm was greater in female rats for the first five 15 minute intervals, but none attained statistical significance (P > 0.05). CBZ pre treatment resulted in reductions in both SWD frequency (Hz) (male, P = 0.003; female, P < 0.0001) and latency to onset of SWD (male, P = 0.002). The frequency of SWD in CBZ pre-treated rats was lower in females (5.8 Hz vs. 6.1 Hz, P = 0.002) as was the decrease in the SWD burst duration following CBZ versus vehicle pre-treatment (-0.05 seconds vs. -0.25 seconds, P = 0.046). CONCLUSIONS: CBZ consistently aggravates absence seizures in the low-dose PTZ model in both female and male rats. However, while some gender differences were found, the results failed to support the hypothesis that females are significantly more susceptible to aggravation of the number or duration of absence seizures by CBZ. PMID- 15121128 TI - Chvostek's sign and hypocalcaemia in children with seizures. AB - Chvostek's sign (CS) was investigated in 154 patients with seizures (epilepsy, n = 91; non-epileptic event, n = 41; febrile convulsion, n = 19; hypocalcaemic seizure, n = 3). Patients with febrile convulsions or non-epileptic seizures had either negative or mild CS. Marked CS was only found among those with the diagnosis of epilepsy or hypocalcaemia. Normocalcaemic patients had no other signs of neuromuscular hyperexcitability (NMH) while those with hypocalcaemia manifested positive Trousseau's sign (TS) and other signs of NMH. There was no significant correlation between CS and seizure control, epilepsy classification and EEG findings. PMID- 15121129 TI - Zonisamide monotherapy in a multi-group clinic. AB - OBJECTIVE: Reports on zonisamide monotherapy are limited despite favourable preliminary data, and typically restricted to tertiary referral centres. The goal of this study is to report clinical experience with zonisamide monotherapy in a large, multi-group clinic setting. METHODS: We reviewed the charts of patients treated with zonisamide monotherapy in the Neurology Department of the Kelsey Seybold Clinic (Houston, Texas) during an 18-month period. We analysed subgroups of patients who were naive to antiepileptic drug (AED) therapy (Group 1) and those who had previous exposure to AEDs (Group 2). RESULTS: The study included 54 paediatric and adult patients with a variety of seizure types: 15 patients in Group 1 and 39 patients in Group 2. Mean maintenance zonisamide dosages in the two groups were similar (193 mg/day in Group 1 vs. 218 mg/day in Group 2). Thirty eight patients (70.4%) continued zonisamide monotherapy, with 7 patients (13.0%) adding a second AED and 9 patients (16.7%) switching to a different drug. Of the 24 patients who became seizure free on zonisamide monotherapy, 11 were on the 100 mg initial dosage. Zonisamide monotherapy was well tolerated. CONCLUSIONS: Zonisamide monotherapy is safe and effective for a variety of seizure types and may be appropriate as first-line therapy in some cases. PMID- 15121130 TI - Epileptic negative myoclonus in a case of atypical benign partial epilepsy of childhood: a detailed video-polygraphic study. AB - PURPOSE: To describe the clinical and electroencephalographic features of a child diagnosed as having atypical benign partial epilepsy (ABPE) who suffered from frequent lapses of postural tone in the right lower limb that were considered to represent a focal epileptic negative myoclonus (ENM). MATERIAL AND METHODS: Electrophysiological evaluation included four serial waking and two sleep EEGs. Moreover, the investigation included a polygraphic recording with simultaneous video-EEG monitoring performed in rest, during hyperventilation, and while standing up with the aim of capturing patient's typical seizures. RESULTS: During awake the EEG showed frequent bilateral centrotemporal discharges maximal over the left central area. Moreover, we recorded short generalised paroxysms of spike and-wave discharges accompanied by a clear transient cognitive impairment that were in keeping with absences. We also captured several episodes of sudden and unexpected loss of postural tone in the right lower limb. These focal inhibitory seizures were associated with brief diffuse but asymmetrical paroxysms of irregular spike-and-wave discharges maximal over the left side. EMG flattening of the right quadriceps muscle was time-locked to the midline of the slow-wave component of the diffuse spike-and-wave discharges. CONCLUSIONS: ENM leading to focal lapses of postural tone in lower extremities may be a predominant type of seizures in ABPE. A polygraphic recording with video-EEG monitoring is essential to confirm the diagnosis, and to detect other concomitant seizures, such as atypical absences. In our case, ENM could be more likely due to a transient disruption of cortical function. PMID- 15121131 TI - Neuronal ceroid lipofuscinosis: a clinicopathological study. AB - We report the clinical, electrophysiological, radiological and morphological features in a series of 12 patients of histopathologically confirmed cases (infantile, juvenile and adult onset) of neuronal ceroid lipofuscinosis (NCL) observed from 1979 to 1998 at National Institute of Mental Health & Neurosciences (NIMHANS), Bangalore (South India). The commonest type of NCL was juvenile (n = 8, 67%) while infantile and adult forms were two each (n = 2, 16.8%). The age at presentation ranged from 2 to 45 years (mean--12.6, 14.3 years; median--7 years; M:F ratio of 2:1). Four patients (33%) had positive family history and five patients had history of consanguineous parentage (41.6%). The commonest presenting symptoms were regression of milestones (83.3%) and/or seizures, myoclonus (83.8%) followed by involuntary choreiform movements (50%), visual loss (41.6%), ataxia (33.3%) and abnormal behaviour (16.6%). Neuro-ophthalmological abnormalities like optic atrophy (50%), macular degeneration (33.3%) and retinitis pigmentosa (8.3%) were seen in two thirds. Nerve conduction studies (n = 4) revealed abnormalities in two, suggestive of sensorimotor neuropathy. Scalp EEG (n = 9) showed slowing of background activity (BGA) of varying degrees with paroxysmal bursts of seizure discharges in majority. Cranial CT scan (n = 4) revealed varying degrees of diffuse atrophy. Diagnostic brain biopsy was carried out in 11 and brain was examined at autopsy in 1 case. Histological examination revealed characteristic PAS and Luxol Fast Blue (LFB) positive, autofluorescent (AF) intracellular ceroid material, both in neurons and astrocytes in the grey matter. Electron microscopy (n = 5) revealed curvilinear (n = 4), lamellar (n = 2) and electron dense (n = 2) inclusions in neurons, astrocytes and vascular endothelial cells. To conclude, this neurodegenerative disease had varied but characteristic clinical presentations and required histopathological confirmation of diagnosis. PMID- 15121132 TI - Topiramate efficacy in an infant with partial seizures refractory to conventional antiepileptic drugs. AB - Many studies showed that Topiramate (TPM) may be a useful drug in a wide spectrum of childhood epilepsies. We report a 3-month-old female with stormy onset of secondarily generalized partial seizures. She showed a high seizure frequency and a progressive worsening electroencephalogram (EEG), despite standard antiepileptic drugs administration. TPM succeeded in controlling seizures, even after the other drugs were discontinued. This case suggests that TPM may represent a good choice for the treatment of partial seizures refractory to conventional drugs in infants. PMID- 15121133 TI - Epidemiology and management of epilepsy in Hong Kong: an overview. AB - Over half of the estimated 50 million people with epilepsy live in Asia, but there has been limited information on the epidemiology, aetiology and management of epilepsy from this region. In this article, we summarise some of the main problems faced by patients and the current treatment options available in an urban area of China. PMID- 15121134 TI - Simple partial seizures (isolated auras) in medial temporal lobe epilepsy. AB - We analysed whether the medial temporal lobe epilepsy (MTLE) with isolated auras makes any difference in the clinical picture in comparison with MTLE in which only complex partial seizures (CPS) occur. We included 100 patients (aged 16-59 years) with unilateral medial temporal lobe lesions who consecutively underwent presurgical evaluation due to intractable MTLE and who became completely aura- and seizure-free after the anterior temporal resection. Preoperatively, isolated auras were present in 70 patients. These patients were categorised into the IA group. The remaining 30 patients in whom the auras preceded seizures were categorised into the NIA group. We found no difference between the two groups for the age at onset, epilepsy duration or aura types. Conversely, a right-sided epileptogenic region (61%) occurred more frequently in the IA group than in the NIA group (27%, P = 0.001). Conclusively, isolated auras show affinity to the right hemisphere. One explanation may be that seizures stop more quickly in the right hemisphere. Another hypothesis is that consciousness can be disturbed much easier by the ictal activity in left temporal seizures: auras evolve more frequently to CPS due to the disturbance of consciousness. PMID- 15121135 TI - Single-photon emission computed tomography in a patient with ictal metamorphopsia. AB - Metamorphopsia is a type of visual illusion, which has been reported as a rare ictal manifestation. The patient presented with a simple partial status epilepticus characterised by continuous facial metamorphopsia, intermittently accompanied by elementary visual hallucinations or other types of visual illusions. Subtraction single-photon emission computed tomography images showed an increased perfusion in the ventrolateral aspect of the right temporo-occipital junction (middle and inferior occipital, and inferior temporal gyri). The result suggests that the anatomical substrate involved in the generation of ictal facial metamorphopsia is located in the visual association areas at the right temporo occipital junction. PMID- 15121136 TI - Immediate (overnight) switching from carbamazepine to oxcarbazepine monotherapy is equivalent to a progressive switch. AB - This study compared immediate (overnight) and progressive switching to oxcarbazepine monotherapy in patients with partial seizures unsatisfactorily treated with carbamazepine monotherapy. Patients were randomised to either an overnight (n = 140) or a progressive switch (n = 146) from carbamazepine to oxcarbazepine monotherapy at a dose ratio of 1:1.5. The difference between the two switch groups in the mean monthly seizure frequency supported the equivalence of overnight and progressive switching (difference of 0.02 excluding outliers; 95% confidence interval (CI) -0.74, 0.78). Following the switch from carbamazepine to oxcarbazepine, there was a reduction in median monthly seizure frequency in both the overnight group (from 1.5 to 0; P = 0.0005) and the progressive group (from 1.0 to 0.4; P = 0.003). The proportion of seizure-free patients increased from 38 to 51% (P = 0.002) and 39 to 49% (P = -0.01) in the overnight and progressive groups, respectively. In addition, the proportion of patients experiencing no clinically significant adverse events did not differ between the two switch methods (difference of 2.5; 95% CI -4.1, 9.0). For patients who are unsatisfactorily treated with carbamazepine monotherapy, overnight switch to oxcarbazepine monotherapy is as effective and well tolerated as a progressive switch, therefore allowing simple and flexible individualised treatment. Switching to oxcarbazepine monotherapy appears to be beneficial for patients who are unsatisfactorily treated with carbamazepine monotherapy, independently of the switch method used. PMID- 15121137 TI - Non-ketotic hyperglycaemia presenting as epilepsia partialis continua. AB - Epilepsia partialis continua (EPC) is a rare epileptic syndrome, observed in various cortical lesions and also in metabolic disorders. We report the case of a 57-year-old patient with EPC as the first manifestation of hyperosmolar non ketotic hyperglycaemia (NKH) of diabetes mellitus. Computed tomography (CT) and magnetic resonance imaging (MRI) were normal. Initial laboratory data revealing serum glucose 1540 mg/dl, and serum osmolality 391 mOsm/l confirmed the diagnosis. Electroencephalography (EEG) showed ictal discharges in the ipsilateral hemisphere during focal seizures. Seizures are resistant to anticonvulsant treatment and respond best to insulin and rehydration. Focal motor seizures or EPC are commonly a symptom of NKH in the elderly patients. We recommend that in such cases a metabolic disorder such as diabetes mellitus be ruled out. PMID- 15121138 TI - Generalised paroxysmal fast activity (GPFA) is not always a sign of malignant epileptic encephalopathy. AB - PURPOSE: Generalised paroxysmal fast activity (GPFA) consists of 8-26 (most frequently around 10 Hz), 2-50 seconds (usually below 10 seconds) bursts of generalised rhythmic discharges with frontal predominance, appearing most frequently during NREM sleep. The pattern is traditionally linked to Lennox Gastaut (LGS) or late LGS (LLGS) syndrome and associated with tonic-axial seizures, pharmaco-resistency and poor prognosis including mental deterioration. We present here four patients with GPFA, who had neither LGS/LLGS syndrome, nor mental deterioration, two of them are seizure-free, two had infrequent seizures on medication. METHODS: All the patients had neurological work-up and high resolution MRI studies. They were followed for years including repeated EEGs in awake state and during sleep. RESULTS: The main seizure-type was generalised tonic-clonic seizure in three patients and absence in one. Onset of epilepsy varied from 7 to 21 years. No MRI lesion was found. All of them had generalised spike-and-wave discharges during the course of their epilepsy but some had also focal clinical or EEG features, were more difficult to treat, as atypical features compared to the classic generalised epilepsies with generalised spike wave pattern. CONCLUSIONS: Although GPFA should remain an important diagnostic feature of both the classical and late variant of LGS, our cases clearly demonstrate that GPFA could represent a possible electrographic variant in certain generalised epilepsies showing atypical features; better treatability and outcome than in LGS and no mental deterioration. PMID- 15121139 TI - The role of seizures in reversible posterior leukoencephalopathy. AB - We report two hypertensive females, one suffering from chronic renal failure and on regular dialysis, the other with eclampsia. Both developed new onset seizures. The patients' MRI of brain showed signals consistent with the reversible posterior leukoencephalopathy syndrome (RPLES), but with the signals seen only in the hemisphere where the seizures predominated. The anatomic correlation of the clinical and imaging findings supports the notion that seizures have a major role in the genesis and evolution of RPLES. PMID- 15121140 TI - Seizures induced by nursery rhymes and children's games. AB - We report the case of a 20 years aged male patient with seizures induced by nursery rhymes and children's games. Seizures were precipitated by various triggers, including thinking to a children's rhyme, to a children's game, to the action of giving a kiss with a hand. Among the above triggers, only the last one was able to induce a seizure during our observation, characterised by jerking of the upper limbs and loss of consciousness with a quick recovery. The electroclinical features were of a brief paroxysm and diffuse theta/delta activity on electroencephalogram, with frontal maximal expression. The playful aspect and the reference to childhood intrinsically associated with an emotional component seem to be the true feature all the stimuli have in common and therefore this form could be classified as an emotional one. To our knowledge seizures precipitated by these kind of stimuli have never before been reported. PMID- 15121141 TI - Heavy coffee drinking and epilepsy. AB - Dietary habits have been rarely associated with seizure frequency in patients with epilepsy. We report a case of a man with a partial symptomatic epilepsy whose daily habit of heavy coffee drinking was associated with an increased seizure frequency. This patient witnessed a dramatic decrease in the frequency of his seizures after stopping coffee ingestion. Caffeine is a global stimulant and the reduction of its intake may help in the treatment of epilepsy. PMID- 15121142 TI - Pharmacology and pharmacokinetics of sedatives and analgesics. AB - The agents used for sedation and analgesia during endoscopy have complex pharmacokinetic and pharmacodynamic properties. Knowledge of these characteristics is necessary for determining the proper agent and dose for specific patient needs. Short-acting agents, such as fentanyl, midazolam, and propofol, provide rapid sedation with a short duration of action that allows patients to return to normal functioning rapidly. When designing a dosing regimen with these agents, age and organ (liver, kidney) function of patients and concomitant medications that may interfere with metabolic and elimination pathways must be considered. PMID- 15121143 TI - Efficacy, safety, and limitations in current practice of sedation and analgesia. AB - The ease and availability of endoscopy as a diagnostic and therapeutic modality for gastrointestinal disorders has greatly increased the number of procedures performed in the United States. One of the main factors in achieving a flawless procedure is the use of sedation and analgesia in endoscopy. This article examines the efficacy, safety, and limitations inherent in the current practice of sedation and analgesia. PMID- 15121144 TI - Update and review of moderate and deep sedation. AB - The administration of sedation/analgesia to allow patients to tolerate diagnostic or therapeutic procedures and tests is a fact of life. Approximately 200,000,000 such sedations are administered every year in the United States. Any morbidity or mortality associated with sedation is likely to be deemed avoidable. There will never be adequate resources or personnel so that trained anesthesia providers administer all sedations, and there probably is no need for such an approach. Clinicians from many specialties now safely and effectively administer sedation. With attention to updated guidelines and concepts, as outlined in this article, the safety and efficacy of sedation/analgesia can and should be excellent. PMID- 15121145 TI - Propofol: a gastroenterologist's perspective. AB - The use of propofol for GI endoscopy has left the realm of experimentation and is now a viable alternative to standard sedation and analgesia. In the hands of appropriately trained gastroenterologists and registered nurses, propofol has been shown to be superior to standard sedation and analgesia in terms of patient satisfaction and comfort and shorter recovery parameters. Comparative studies have found it to be as safe as the regimens that are used for standard sedation and analgesia. Its narrow therapeutic window demands that specially trained personnel who are not directly involved in the endoscopic procedure administer it. Cost-effectiveness data suggest that propofol is superior to conventional sedation and analgesia, even with the use of added personnel.The importance of pre-procedural assessment and appropriate monitoring cannot be overemphasized. The endoscopist must have a thorough knowledge of the pharmacology of the agents used for sedation and the training necessary to recognize and manage over sedation. Numerous regulatory groups are carefully scrutinizing the practice of sedation and analgesia. It seems that ventilatory monitoring will be required for at least a subset of patients. Although hypercapnia and apnea can be reliably measured, the most important questions to be answered are if such monitoring affects patient outcomes and which patients are at risk for apnea and alveolar hypoventilation. PMID- 15121146 TI - A nursing perspective on sedation and nurse-administered propofol for endoscopy. AB - Although the role of nurses in endoscopic sedation and monitoring varies widely, our experience has been that an active role of nurses in the administration of sedation and in patient monitoring has been safe for patients, has allowed expansion of the use of propofol in GI endoscopy in a cost-effective fashion, and has been satisfying to patients and nurses. In addition, in some instances nurse administration of sedation has improved the efficiency of the endoscopy unit. The role of nurses in endoscopic sedation and monitoring continues to develop, and we are enthusiastic about an active role for nurses. PMID- 15121147 TI - Extended/advanced monitoring techniques in gastrointestinal endoscopy. AB - The practice of sedation and analgesia is under increasing scrutiny by numerous regulatory agencies, with the aim of making these procedures safer and reducing the incidence of cardiopulmonary complications during GI endoscopy. As we move toward more evidence-based medicine, new technologies will have to be assessed in a manner that demonstrates their efficacy and utility in clinical practice. Although there have been no controlled studies examining whether more intensive monitoring during endoscopy improves outcomes, extended monitoring with capnography seems to offer an advantage over conventional monitoring in that, by providing a real-time indication of any change in adequate ventilation before oxygen desaturation occurs, it can detect early phases of respiratory depression, which can allow a more precise and safer titration of medications. There is a close agreement among experts that capnography may reduce the risk of adverse outcomes during deep sedation; therefore, its use should be required for patients undergoing advanced endoscopic procedures with the potential for deep sedation. Extended monitoring with capnography should also be endorsed whenever propofol is considered as an alternative to standard sedation with a benzodiazepine or narcotic. Our understanding of the clinical application of techniques for monitoring of depth of sedation is in its infancy, and its full contribution to the practice of endoscopy has yet to be determined. Their potential role in improving sedation practice during endoscopy needs to be confirmed by controlled trials. If we consider the lack of proven efficacy of these emerging monitoring techniques in reducing the adverse outcomes associated with sedation and analgesia, the importance of appropriate monitoring cannot be overemphasized. However, it is vital for the endoscopist to be thoroughly familiar with the type of sedation chosen, to be able to recognize the various levels of sedation, and, above all, to rescue patients should they unintentionally progress to a deeper level of sedation than intended. PMID- 15121148 TI - Patient-controlled analgesia and sedation in gastrointestinal endoscopy. AB - Variations in pain threshold, drug tolerance, and visceral sensitivity among patients make it difficult to anticipate the appropriate dose of sedation for gastrointestinal endoscopy. Propofol was recently introduced for sedation in endoscopy and has a rapid onset and offset of action, making it an ideal substance for patient-controlled administration. Several controlled trials have demonstrated that during colonoscopy, patient-controlled application of propofol alone or in combination with various opioids is effective,safe, and yields high patient satisfaction. Target-controlled infusion of propofol has shown encouraging results for prolonged upper endoscopy procedures like endoscopic retrograde cholangio pancreatography. PMID- 15121149 TI - Unsedated endoscopy. AB - Unsedated endoscopy will likely play an increasingly important role in the daily practice of GI endoscopy. Although there is adequate evidence that un-sedated endoscopy is technically feasible in selected patient populations, there area number of obstacles preventing its widespread adoption. Small-caliber endoscopy is not as accurate as conventional EGD. This may be remedied as improvements in endoscope technology emerge. Many patients refuse to consider an unsedated examination, and those who do participate report more symptoms of discomfort than patients undergoing conventional endoscopy. Whether patients will accept mild discomfort in exchange for substantial cost savings has not been evaluated. Finally, although unsedated endoscopy seems to have a low complication rate, its safety has not been formally addressed in large studies. Large, randomized, controlled studies are needed to better determine the role of un-sedated small caliber endoscopy in daily practice. Unsedated colonoscopy may follow the footsteps of unsedated upper endoscopy with even more obstacles to overcome before its widespread application. PMID- 15121150 TI - Sedation and analgesia in the pediatric patient. AB - The administration of sedation and analgesia for pediatric gastrointestinal procedures has become routine but is not standardized. For the most part, pediatric endoscopists are encouraged to use their clinical judgment to select between using intravenous (IV) sedation or general anesthesia on an individual patient basis. Commonly administered IV sedation regimens in children combine benzodiazepines with narcotics, but anesthesiologist administered propofol sedation is gaining acceptance among pediatric gastroenterologists. Guidelines for patient monitoring and new technologic advances may help to ensure patient safety for children undergoing endoscopic procedures, no matter what sedation regimen is used. PMID- 15121151 TI - Risk management regarding sedation/analgesia. AB - Because the definition of standard of care changes with time, it is difficult to predict whether any given technique or practice will be ultimately viewed as the next advance in care or will be discarded as unnecessary or cumbersome. It is too early to tell whether deep sedation/analgesia with propofol or some other ultrashort acting anesthetic will emerge as the sedative/analgesic of choice for routine endoscopic practice. It seems that deep sedation with propofol provides some advantages over current narcotic/benzodiazepine combinations in many circumstances, but potential risks exist. Further information, including careful documentation of the complications that occur with this and similar agents, be they attributable to the drug or the level of sedation, are eagerly awaited. For now, it seems prudent to continue to recommend that the level of sedation/analgesia be appropriate to the perceived need and that patients continue to have their procedures performed with safety, comfort, and competence as the overriding concerns. PMID- 15121152 TI - Quality assurance in the endoscopy suite: sedation and monitoring. AB - Education and training is the first line of defense in maintaining patient safety and providing quality care in the Endoscopy suite. Ensuring that the health care provider is well trained minimizes the risk to the patient. Quality assurance begins and ends with training. Quality assurance in the endoscopy suite related to sedation is accomplished through training and ongoing data collection using structure, process, and outcome measures. Monitoring compliance to standards with continual feedback of results allows the endoscopist to evaluate performance on an ongoing basis. The endoscopist has no choice but to become an active participant in the quality assurance process to improve the quality and value of their work. PMID- 15121155 TI - Studying cardiac arrhythmias in the mouse--a reasonable model for probing mechanisms? AB - The normal mechanical functioning of the heart depends on proper electrical functioning, reflected in the sequential activation of pacemaker cells, and the normal propagation of activity through the ventricles. Myocardial electrical activity is evident in the form of action potentials, reflecting the activation (and inactivation) of depolarizing (Na(+), Ca(2+)) and repolarizing (K(+)) current channels. There are multiple types of myocardial K(+) channels, contributing to regional differences in action potential waveforms and to the generation of normal cardiac rhythms. The conduction and propagation of activity through the myocardium depends on electrical coupling between cells, mediated by gap junction channels. In the diseased myocardium, action potential waveforms and conduction are affected markedly, owing to changes in the functional expression of repolarizing K(+) and other channels. These changes can lead to desynchronization of the heart and to arrhythmia generation. There is presently greater interest in defining the cellular, molecular, and systemic mechanisms contributing to the generation and the maintenance of cardiac arrhythmias. Although a variety of experimental (animal) model systems have been (and are being) exploited in these efforts, the mouse is being used increasingly, due to the ease with which molecular genetic strategies can be applied. The important issue is whether the mouse is an appropriate model system to explore arrhythmia mechanisms. PMID- 15121156 TI - GPCR expression in the heart; "new" receptors in myocytes and fibroblasts. AB - G protein-coupled receptor (GPCR) signaling cascades play a key role in regulation of the cardiovascular system. GPCRs comprise the largest cell-surface receptor superfamily and are also the largest class of drug targets to which agonists and antagonists are currently directed. This article reviews recent data regarding aspects of GPCR signaling and emphasizes the recent "discovery" of expression of "new" GPCRs in cardiac myocytes and fibroblasts. In addition to several well-known ("classical") GPCRs that are expressed by cardiac myocytes and fibroblasts, these cell types or cardiac tissue (cell types as-yet undefined) express a large number of other GPCRs, including receptors for peptides, lipids, and other molecules, as well as "orphan" receptors for which the natural agonists and (patho)physiologic roles in the cardiovascular system have not yet been defined. Studies of the newly recognized GPCRs-in particular, deorphanization of novel GPCRs and experiments that define their signal pathways, expression patterns, desensitization mechanisms, cross-talk with other receptors, and genetic variation-are likely to yield important new insights into cardiovascular cell function. Moreover, these "new" GPCRs are attractive targets for the development of drugs for the treatment of cardiac and other cardiovascular disorders. PMID- 15121157 TI - MMPs, cadherins, and cell proliferation. AB - Proliferation of vascular smooth muscle cells plays a key role in the development of pathologic processes characterized by intimal thickening. Cadherins are adhesion receptors involved in homophilic cell-cell interactions, but they can also act as signaling molecules that affect cell behavior, including proliferation, migration. and survival. The matrix-degrading metalloproteinases (MMPs) are a family of proteolytic enzymes that modulate tissue remodeling, proliferation, and migration by degradation of extracellular matrix proteins as well as non-matrix proteins, including cadherins. This review discusses the key links between cadherins and MMPs in cell proliferation. A greater understanding of these processes may aid with the design of new clinical therapeutic approaches for atherosclerosis and restenosis. PMID- 15121158 TI - Matrix metalloproteinases and vascular endothelium-mononuclear cell close encounters. AB - Interactions between circulating mononuclear cells and vascular endothelium are key events in the pathogenesis of cardiovascular diseases such as atherosclerosis. Leukocyte adhesion to endothelial cells or to components of the endothelial basement membrane can trigger the synthesis and secretion of matrix metalloproteinases (MMPs), enzymes capable of degrading extracellular matrix. Secretion of MMPs is likely to facilitate leukocyte transmigration into the vascular wall, but may serve other functions as well, which could include providing negative feedback regulation of the inflammatory response. PMID- 15121159 TI - Human genomics and its impact on arrhythmias. AB - Investigation of families with monogenic arrhythmia syndromes has identified genes whose further study has proven extraordinarily valuable in further understanding variability in cardiac electrophysiology and its response to exogenous stressors. One lesson being learned from this work is that individuals from the same kindred with the same disease-associated DNA variant may nevertheless display strikingly different phenotypes. Further, common function altering polymorphisms are being identified in these and other genes controlling cardiac excitability. These findings, combined with the recognition that very common arrhythmia syndromes (such as atrial fibrillation or sudden arrhythmic death) include a genetic component, raise the prospect that analysis of genomic variability among individuals and populations may in the future be used to manage patients PMID- 15121160 TI - Angiotensin II-mediated development of vascular diseases. AB - Angiotensin II (AngII) has well-characterized effects on blood pressure and fluid balance that adversely affect atherosclerotic cardiovascular disease. More recently, there is a realization that AngII exerts direct effects on arterial wall cells to influence atherosclerotic lesion formation. Several groups have shown that infusion of AngII into hyperlipidemic mice rapidly and profoundly augments lesion formation. The increase in lesions from AngII was not attributable to elevated blood pressure. The lesions formed from AngII infusion are overtly similar to those formed during hypercholesterolemia, with infiltration of macrophages and T lymphocytes. Unexpectedly, AngII infusion into these mice also led to the development of abdominal aortic aneurysms. These aneurysms exhibit many aspects of the human disease including medial degeneration, inflammation, thrombus, and rupture. The definition of the cellular mechanisms by which Ang II promotes these vascular pathologies may provide new therapeutic strategies. PMID- 15121161 TI - Heart induction: embryology to cardiomyocyte regeneration. AB - Developmental biologists have uncovered many of the signaling proteins that are required to recruit early embryonic cells to the myocardial lineage. A detailed understanding of their function should provide insights into the difficult task of inducing embryonic stem cells to develop cardiomyocyte precursors for cell based therapies. These proteins may also prove to be useful to stimulate progenitor cells to differentiate into cardiomyocytes within the damaged heart. PMID- 15121163 TI - Advanced transfection with Lipofectamine 2000 reagent: primary neurons, siRNA, and high-throughput applications. AB - Lipofectamine 2000 is a cationic liposome based reagent that provides high transfection efficiency and high levels of transgene expression in a range of mammalian cell types in vitro using a simple protocol. Optimum transfection efficiency and subsequent cell viability depend on a number of experimental variables such as cell density, liposome and DNA concentrations, liposome-DNA complexing time, and the presence or absence of media components such as antibiotics and serum. The importance of these factors in Lipofectamine 2000 mediated transfection will be discussed together with some specific applications: transfection of primary neurons, high throughput transfection, and delivery of small interfering RNAs. PMID- 15121164 TI - FuGENE 6 Transfection Reagent: the gentle power. AB - FuGENE 6 Transfection Reagent has been commercially available since 1997. Since that time, its popularity has increased due to its ease of use, minimal to no cytotoxicity, and the high level of transfection in many different cell lines. FuGENE 6 Transfection Reagent is gentle on the cells. Adherent cells can be trypsinized and transfected by the DNA:FuGENE 6 reagent complex prior to plating, making it a strong candidate for high throughput applications. Additionally, low cell numbers can be transfected in 96-well plates. As with most reagents, the complex formation step is critical and special handling is required because the reagent is supplied in 80% ethanol. For example, contact with plastic must be avoided as inhibitors of transfection can leach from some plastics. We investigated parameters that have been reported to affect the transfection efficiency including the use of common antibiotics, passage level of the cells, and length of time for complex formation. These parameters are often cell line dependent and can be optimized to increase transfection efficiency for a specific cell line. PMID- 15121166 TI - Biolistic and diolistic transfection: using the gene gun to deliver DNA and lipophilic dyes into mammalian cells. AB - Delivery of genes by firing them into mammalian cells is becoming increasingly popular for cells such as neurones, that have proved difficult to transfect by conventional means. In addition it is becoming apparent that this technique also provides a suitable method of introducing cell-specific dyes into mammalian tissues. Here we describe improved protocols for the rapid and efficient delivery of both DNA and lipophilic dyes into mammalian cells in both monolayers and in brain slices. The results show that transfection of genes, such as that for enhanced yellow fluorescent protein, using a modified microcarrier preparation technique combined with either a conventional or novel accelerator channel, yields rapid and efficient production of protein, which can be visualised in 24 48 h. Following similar delivery of lipophilic dyes neurones can be visualised in minutes. These techniques therefore provide an excellent means not only to examine the structure and function of neurones and neuronal genes, but also those of other cell types. PMID- 15121165 TI - Mouse embryonic stem cells efficiently lipofected with nuclear localization peptide result in a high yield of chimeric mice and retain germline transmission potency. AB - Embryonic stem (ES) cells are an important tool in developmental biology, genomics, and transgenic methods, as well as in potential clinical applications such as gene therapy or tissue engineering. Electroporation is the standard transfection method for mouse ES (mES) cells because lipofection is quite inefficient. It is also unclear if mES cells treated with cationic lipids maintain pluripotency. We have developed a simple lipofection method for high efficiency transfection and stable transgene expression by employing the nonclassical nuclear localization signal M9 derived from the heterogeneous nuclear ribonucleoprotein A1. In contrast to using 20 microg DNA for 10 x 10(6) cells via electroporation which resulted in 10-20 positive cells/mm2, M9-assisted lipofection of 2 x 10(5) cells with 2 microg DNA resulted in > 150 positive cells/mm2. Electroporation produced only 0.16% EGFP positive cells with fluorescence intensity (FI) > 1000 by FACS assay, while M9-lipofection produced 36-fold more highly EGFP positive cells (5.75%) with FI > 1000. Using 2.5 x 10(6) ES cells and 6 microg linearized DNA followed by selection with G418, electroporation yielded 17 EGFP expressing colonies, while M9-assisted lipofection yielded 72 EGFP expressing colonies. The mES cells that stably expressed EGFP following M9-assisted lipofection yielded > 66% chimeric mice (8 of 12) and contributed efficiently to the germline. In an example of gene targeting, a knock-in mouse was produced from an ES clone screened from 200 G418 resistant colonies generated via M9-assisted lipofection. To our knowledge, this is the first report of generation of transgenic or knock-in mice obtained from lipofected mES cells and this method may facilitate large scale genomic studies of ES developmental biology or large scale generation of mouse models of human disease. PMID- 15121167 TI - In vitro and in vivo electric field-mediated permeabilization, gene transfer, and expression. AB - Electropulsation is one of the non-viral methods successfully used to transfer genes into living cells in vitro as in vivo. This approach shows promise in the field of gene and cellular therapies. The present paper first describes the factors controlling electropermeabilization to small molecules (< 4 kDa) and then the processes supporting DNA transfer in vitro. The description of in vitro events brings the attention of the reader to the processes occurring before, during, and after electropulsation of DNA and cells. Their developments for the in vivo processes are reported in the final part where the present and potential clinical applications are described. PMID- 15121168 TI - Transfection of adherent and suspended cells by calcium phosphate. AB - DNA-calcium phosphate coprecipitates have been used for 30 years as an efficient method to introduce genetic material into cells. The method involves simple solutions that can be prepared or purchased by the experimentalist. All the numerous variations of the protocol found in the literature are based on the same principle--a spontaneous precipitation that occurs in supersaturated solutions. When DNA is present during this process, it is readily incorporated into the forming calcium phosphate precipitate. Although a wide range of conditions will lead to precipitates, high transfection efficiencies are only obtained within a narrow range of optimized parameters that assure certain properties of the precipitate. This paper describes several physico-chemical parameters that are critical to adapt the method to a particular cell line and/or cultivation condition. Examples of protocols that were established and tested within the authors' laboratory are presented. The article also emphasizes differences between transfections of adherent and suspended cells. PMID- 15121169 TI - Polyethylenimine strategies for plasmid delivery to brain-derived cells. AB - The introduction of effective transfection reagents has had a dramatic impact on basic scientific studies over the past decade and is methodically becoming a clinical relevant agent. An area where these agents have had little impact to date is in transfection of neuronal cells either in vivo or in vitro. The poor results, obtained with these cells, likely arise from the innate properties of the cell itself such as its post-mitotic state and its fragility to the transfection agent. In this report, we investigated the transfection efficiency of branched and linear form of polyethylenimine (PEI) for a commonly used tissue culture cell line, the human CF bronchial epithelial cell line IB3-1, rat brain derived glial, and neuronal cell lines. In addition, the effect of reaction conditions, such as ratio of PEI/plasmid, polymer molecular weight, and shape, was addressed on the transfection effects. The results indicate that branched PEI is more effective for the brain-derived cells. It is also shown that PEI 25 is more effective for the glial cells and PEI 50-100 is more effective for the neuronal cells under the evaluation conditions. PMID- 15121170 TI - New non-viral method for gene transfer into primary cells. AB - The availability of genetically altered cells is an essential prerequisite for many scientific and therapeutic applications including functional genomics, drug development, and gene therapy. Unfortunately, the efficient gene transfer into primary cells is still problematic. In contrast to transfections of most cell lines, which can be successfully performed using a variety of methods, the introduction of foreign DNA into primary cells requires a careful selection of gene transfer techniques. Whereas viral strategies are time consuming and involve safety risks, non-viral methods proved to be inefficient for most primary cell types. The Nucleofector technology is a novel gene transfer technique designed for primary cells and hard-to-transfect cell lines. This non-viral gene transfer method is based on a cell type specific combination of electrical parameters and solutions. In this report, we show efficient transfer of DNA expression vectors and siRNA oligonucleotides into a variety of primary cell types from different species utilizing the Nucleofector technology, including human B-CLL cells, human CD34+ cells, human lymphocytes, rat cardiomyocytes, human, porcine, and bovine chondrocytes, and rat neurons. PMID- 15121171 TI - Lentiviral and MLV based retroviral vectors for ex vivo and in vivo gene transfer. AB - Retroviral vectors have become an important tool for gene transfer in vitro and in vivo. Classical Moloney murine leukemia virus (MLV) based retroviral vectors have been used for over 20 years to transfer genes into dividing cells. Cell lines for production of retroviral vectors have become commonly available and modifications in retroviral vector design and use of envelope proteins have made the production of high titer, helper-free, infectious virus stocks relatively easy. More recently, lentiviral vectors, another class of retroviruses, have been modified for in vitro and in vivo gene transfer. The ability of lentiviral vectors to transduce non-dividing cells has made them especially attractive for in vivo gene transfer into differentiated, non-dividing tissues. Several improvements in helper plasmids and vectors have made lentivirus a safe vector system for ex vivo and in vivo gene transfer. This review will briefly summarize the background of these vector systems and provide some common protocols available for the preparation of MLV based retroviral vectors and HIV-1 based lentiviral vectors. PMID- 15121172 TI - Utilization of adenovirus vectors for multiple gene transfer applications. AB - Mammalian viruses have evolved over millions of years to achieve a single goal, namely to rapidly enter a host mammalian cell, in order to achieve virus propagation. In so doing, these biologic parasites have acquired the molecular tools to rapidly and efficiently deliver their own nucleic acids into the nucleus of the host cell. The human adenovirus is one of the best studied of these parasites. As such the adenovirus has been re-engineered to allow it to be used as a tool to allow researchers to deliver desired nucleic acid sequences into a large variety of cell targets, both in tissue culture systems, as well as directly into living animals. Adenovirus based gene transfer systems can overcome most of the problems inherent to high efficiency gene transfer, and perform in a fashion that in many ways cannot be matched by most other currently utilized gene transfer systems. This article will attempt to summarize the multiple attributes of this widely utilized gene delivery system. PMID- 15121173 TI - Targeting HSV amplicon vectors. AB - Several techniques have been developed to deliver DNA directly into mammalian cells, spanning in difficulty from simple mixing procedures to complex systems requiring expensive equipment. Viral vectors have proven able to deliver genes into mammalian cells with high efficiency and low toxicity. In particular, herpes simplex virus type-1 (HSV-1) amplicon vectors are well suited for gene transfer studies as they can infect many cell types, both non-dividing and dividing, have a large transgene capacity and are easy to manipulate. For some applications, it may be desirable to target gene delivery to specific cell populations or to transduce normally non-susceptible cells. This can be achieved by modifying one or more of the glycoproteins found in the viral envelope. Glycoprotein C (gC) has a well-characterized heparan sulfate binding domain (HSBD) necessary for HSV binding to cells. Replacing this region with unique ligands can result in less efficient binding to natural target cells and increase binding to cells which express receptors for these ligands. A method to retarget amplicon vectors by replacing gC HSBD with a model ligand, the hexameric histidine-tag, is described, as well as means to evaluate the binding of modified vector as compared to wild type virus to cells with or without the appropriate receptor, in this case, a his tag pseudo-receptor. This protocol demonstrates increased binding of modified virus to receptor-positive cells (at levels greater than wild-type) with no loss of infectivity. Retargeted vectors can provide an additional tool for increasing the efficiency of gene delivery to specific cell types. PMID- 15121174 TI - How to keep injured CNS neurons viable--strategies for neuroprotection and gene transfer to retinal ganglion cells. PMID- 15121175 TI - A dimeric version of the short N-cadherin binding motif HAVDI promotes neuronal cell survival by activating an N-cadherin/fibroblast growth factor receptor signalling cascade. AB - The HAVDI and INPISGQ sequences have been identified as functional binding motifs in extracellular domain 1 (ECD1) of N-cadherin. Cyclic peptides containing a tandem repeat of the individual motifs function as N-cadherin agonists and stimulate neurite outgrowth. We now show that the cyclic peptide N-Ac CHAVDINGHAVDIC-NH2 (SW4) containing the HAVDI sequence in tandem is efficacious also in promoting the in vitro survival of several populations of central nervous system neurons in paradigms where fibroblast growth factor-2 (FGF-2) is active. SW4 supported the survival of rat postnatal cerebellar granule neurons plated in serum-free medium and limited the death of differentiated granule neurons induced to die by switch to low K+ medium. In addition, SW4 rescued embryonic hippocampal and cortical neurons from injury caused by glutamic acid excitotoxicity. The neuroprotective effects of SW4 displayed a concentration dependence similar to those inducing neuritogenesis, were inhibited by a monomeric version of the same motif and by a specific FGF receptor antagonist (PD173074), and were not mimicked by the linear peptide. Inhibitors of the phosphatidylinositol 3-kinase (PI 3 kinase), MAP kinase, and p38 kinase signalling pathways did not interfere with SW4 function. These data suggest that SW4 functions by binding to and clustering N-cadherin in neurons and thereby activating and N-cadherin/FGF receptor signalling cascade, and propose that such agonists may represent a starting point for the development of therapeutic agents promoting neuronal cell survival and regeneration. PMID- 15121176 TI - Coordinated functions of Netrin-1 and Class 3 secreted Semaphorins in the guidance of reciprocal septohippocampal connections. AB - An essential characteristic of the CNS function is the formation of reciprocal connections between brain areas. Although the mechanisms controlling the establishment of neuronal connections are being determined, very little is known about the development of reciprocal connections, which often course along identical pathways. Here, we show that Netrin-1, expressed along the fimbria, chemoattracts both septohippocampal and hippocamposeptal fibers. Moreover, we show that both Semaphorins 3A and 3F expressed in regions nearby the septum prevent the growth of septal axons into these regions. Blocking experiments with recombinant ecto-Neuropilins indicate that both Semaphorins 3A and 3F act cooperatively in the repulsion of septal axons. Furthermore, netrin-1-deficient mice develop a reduced septohippocampal projection. We conclude that the coordinated actions of Netrin-1 and Semaphorins 3A and 3F cooperate in the development of septohippocampal and hippocamposeptal connections, indicating that the same molecular cues serve the construction of reciprocal connections in both directions of growth. PMID- 15121177 TI - Regulation of Nogo and Nogo receptor during the development of the entorhino hippocampal pathway and after adult hippocampal lesions. AB - Axonal regeneration in the adult CNS is limited by the presence of several inhibitory proteins associated with myelin. Nogo-A, a myelin-associated inhibitor, is responsible for axonal outgrowth inhibition in vivo and in vitro. Here we study the onset and maturation of Nogo-A and Nogo receptor in the entorhino-hippocampal formation of developing and adult mice. We also provide evidence that Nogo-A does not inhibit embryonic hippocampal neurons, in contrast to other cell types such as cerebellar granule cells. Our results also show that Nogo and Nogo receptor mRNA are expressed in the adult by both principal and local-circuit hippocampal neurons, and that after lesion, Nogo-A is also transiently expressed by a subset of reactive astrocytes. Furthermore, we analyzed their regulation after kainic acid (KA) treatment and in response to the transection of the entorhino-hippocampal connection. We found that Nogo-A and Nogo receptor are differentially regulated after kainic acid or perforant pathway lesions. Lastly, we show that the regenerative potential of lesioned entorhino hippocampal organotypic slice co-cultures is increased after blockage of Nogo-A with two IN-1 blocking antibodies. In conclusion, our results show that Nogo and its receptor might play key roles during development of hippocampal connections and that they are implicated in neuronal plasticity in the adult. PMID- 15121178 TI - Weak synaptic activity induces ongoing signaling to the nucleus that is enhanced by BDNF and suppressed by low-levels of nicotine. AB - The developing nervous system adapts to a wide array of stimuli, in part, by evoking activity-dependent mechanisms that signal to the nucleus and induce long term modifications in neuronal function. It is well established that one such stimulus is strong synaptic activity. Our interest, however, is whether weak activity generated at developing synapses also signals to the nucleus and if so, can these signals be modulated by extrinsic factors. Using cultured hippocampal neurons and a highly sensitive readout of CRE-mediated gene expression, we demonstrate that weak synaptic transmission, including non-evoked, spontaneous transmitter release, induces ongoing gene expression. These weak synaptic stimuli, acting through NMDA receptors, signal to the nucleus through a MAPK pathway, without a significant contribution of L-type Ca2+ channels. In addition, we show that BDNF, a molecule that has clear effects on synaptic plasticity, enhances this CRE-dependent gene expression by acting upstream of NMDA receptors. On the other hand, low levels of nicotine, which also effects synaptic plasticity, suppress ongoing CRE-mediated gene expression indirectly by acting on GABAergic neurons; this indirect action on gene expression suggests an alternative mechanism for how nicotine produces long-lasting changes. PMID- 15121179 TI - Excess Bcl-XL increases the intrinsic growth potential of adult CNS neurons in vitro. AB - The regenerative potential of adult mammalian CNS neurons is limited. Recent data suggest that inactivation of major growth inhibitors may not suffice to induce robust regeneration from mature neurons unless the intrinsic growth state is modulated. To investigate a possible role of Bcl-XL for axon regeneration in the adult mammalian CNS, Bcl-XL was adenovirally overexpressed in severed rat RGCs. Bcl-XL overexpression in mature axotomized RGCs in vivo increased both numbers [3.10-fold (+/-0.20)] and cumulative length [6.72-fold (+/-0.47)] of neurites regenerated from retinal explants, and this effect was further pronounced in the central retina where specific and dense axoplasmatic transduction occurs. Similarly, delayed Bcl-XL gene transfer to explanted retinae 12-13 days after lesion increased the numbers and length of emanating neurites by a factor of 5.22 (+/-0.41) and 8.29 (+/-0.69), respectively. In vivo, intraretinal sprouting of unmyelinated RGC axons into the nerve fiber layer was increased. However, fiber ingrowth into the optic nerve remained sparse, likely due to myelin inhibitors and scar components. Therefore, Bcl-XL overexpression may enhance, but may not be sufficient to, restitute functional regeneration in the adult CNS. As assessed by cell quantification analysis, Bcl-XL overexpression rescued a higher proportion of RGCs in vivo than in vitro. Therefore, Bcl-XL is capable to induce both neuronal survival and axon regeneration, but these two processes appear to be differentially modified by distinct pathways in vivo. PMID- 15121181 TI - Semaphorin3A-induced receptor endocytosis during axon guidance responses is mediated by L1 CAM. AB - During axon navigation, Semaphorin3A-induced growth cone retraction is correlated with endocytosis. Although its function remains elusive, we showed previously that the cell adhesion molecule of the immunoglobulin super family L1 associates with Neuropilin-1 (NP-1) the Sema3A-binding subunit of the receptor complex and is required for Sema3A to elicit axonal repulsive responses. We report here that upon Sema3A binding to NP-1, L1 and NP-1 are co-internalized through a clathrin dependent mechanism mediated by L1. We show that in COS7 cells, L1/NP-1 endocytosis is correlated with a cell contraction similar to that observed with the Plexin (Plex)/NP-1 or Plex/NP1/L1 complexes. In neuronal cultures, a L1 mimetic peptide able to switch Sema3A repulsive responses to attraction blocks both endocytosis and growth cone collapse. Similarly, in the COS7 cell model, peptide application prevents both the Sema3-induced L1/NP-1 internalization and cell collapse. These studies demonstrate that the L1/NP-1 complex is able to confer a biological response to Sema3A with L1 mediating receptor internalization following ligand activation. They also reveal that endocytosis controlled by L1/NP-1 cis and trans interactions is pivotal in Sema3A-mediated axon guidance. PMID- 15121180 TI - HB-GAM inhibits proliferation and enhances differentiation of neural stem cells. AB - Proliferation of neural stem cells in the embryonic cerebral cortex is regulated by many growth factors and their receptors. Among the key molecules stimulating stem cell proliferation are FGF-2 and the FGF receptor-1. This ligand-receptor system is highly dependent on the surrounding heparan sulfates. We have found that heparin-binding growth-associated molecule (HB-GAM, also designated as pleiotrophin) regulates neural stem cell proliferation in vivo and in vitro. Deficiency of HB-GAM results in a pronounced, up to 50% increase in neuronal density in the adult mouse cerebral cortex. This phenotype arises during cortical neurogenesis, when HB-GAM knockout embryos display an enhanced proliferation rate as compared to wild-type embryos. Further, our in vitro studies show that exogenously added HB-GAM inhibits formation and growth of FGF-2, but not EGF, stimulated neurospheres, restricts the number of nestin-positive neural stem cells, and inhibits FGF receptor phosphorylation. We propose that HB-GAM functions as an endogenous inhibitor of FGF-2 in stem cell proliferation in the developing cortex. PMID- 15121182 TI - Effects of Wnt1 signaling on proliferation in the developing mid-/hindbrain region. AB - The secreted glycoprotein WNT1 is expressed in the caudal midbrain and is essential for proper development of the entire mid-/hindbrain region. To get better insights into Wnt1 function in the mid-/hindbrain region, we ectopically expressed Wnt1 under the control of the endogenous En1 promoter, thereby extending Wnt1 expression rostrally into the anterior midbrain and caudally into rhombomere 1. In these transgenic mice, the position of the mid-/hindbrain organizer is not altered and pattern formation is not changed. During midgestation, ectopic Wnt1 induced strong overproliferation of precursor cells only in the caudal midbrain in a gene dosage-dependent manner. Enhanced proliferation is at least in part mediated by shortening of the cell cycle length. In adults, Wnt1 exhibited a cell size promoting effect specifically on neurons. We suggest that Wnt1 acts as a regulator of proliferation of specific precursor populations in the developing mid-/hindbrain region and is only secondarily involved in maintenance of the mid-/hindbrain organizer. PMID- 15121183 TI - Disrupted in Schizophrenia 1 (DISC1) is a multicompartmentalized protein that predominantly localizes to mitochondria. AB - DISC1 is disrupted by a chromosomal translocation cosegregating with schizophrenia and recurrent major depression in a large Scottish family and has also been reported as a potential susceptibility locus in independent populations. We reveal a widespread and complex pattern of DISC1 expression, with at least five forms of Disrupted in Schizophrenia 1 DISC1 detectable. Mitochondria are the predominant site of DISC1 expression with additional nuclear, cytoplasmic, and actin-associated locations evident. Although the subcellular targeting of DISC1 is clearly complex, the association with mitochondria is of interest as many mitochondrial deficits have been reported in schizophrenia and other neuropsychiatric illnesses. Moreover, of the many cellular functions performed by mitochondria, their role in oxidative phosphorylation, calcium homeostasis, and apoptosis may hold particular relevance for the neuronal disturbances believed to be involved in the pathogenesis of schizophrenia. PMID- 15121184 TI - Gating and modulation of presumptive NaV1.9 channels in enteric and spinal sensory neurons. AB - The NaV1.9 subunit is expressed in nociceptive dorsal root ganglion (DRG) neurons and sensory myenteric neurons in which it generates 'persistent' tetrodotoxin resistant (TTX-R) Na+ currents of yet unknown physiological functions. Here, we have analyzed these currents in details by combining single-channel and whole cell recordings from cultured rat DRG and myenteric neurons. Comparison of single channel with whole-cell data indicates that recording using internal CsCl best reflects the basic electrical features of NaV1.9 currents. Inclusion of fluoride in the pipette solution caused a negative shift in the activation and inactivation gates of NaV1.9 but not NaV1.8. Fluoride acts by promoting entry of NaV1.9 channels into a preopen closed state, which causes a strong bias towards opening and enhances the ability of sensory neurons to sustain spiking. Thus, the modulation of the resting-closed states of NaV1.9 channels strongly influences nociceptor excitability and may provide a mechanism by which inflammatory mediators alter pain threshold. PMID- 15121185 TI - Embryonic stem cells expressing expanded CAG repeats undergo aberrant neuronal differentiation and have persistent Oct-4 and REST/NRSF expression. AB - Nine neurodegenerative disorders are caused by CAG/polyglutamine (polyQ) repeat expansions. The molecular mechanisms responsible for disease-specific neurodegeneration remain elusive. We developed an embryonic stem (ES) cell-based model to probe the role of polyQ tract expansion in neuronal degeneration. ES cells containing expanded CAG repeats in the hypoxanthine phosphoribosyltransferase (Hprt) gene develop features typical of CAG-mediated neuropathology, exhibit length-dependent decrease in survival, undergo aberrant neuronal differentiation as well as persistent Oct-4 and Repressor element-1 transcription factor/neuron restrictive silencer factor (REST/NRSF) expression. This novel model will allow analysis of the molecular pathogenesis of neuronal degeneration and can be used to rapidly screen therapeutic interventions for these fatal diseases. PMID- 15121186 TI - Identification and characterization of novel properties of the human D3 dopamine receptor. AB - Among dopamine receptors, the function and properties of the D3 receptor subtype are poorly understood. Here we report the identification and characterization of two unique properties of the human D3 receptor. The D3 receptor exhibits a tolerance property wherein the magnitude of the second agonist-induced response is reduced by 60% compared to the first response and progressively decreases upon repeated agonist application. In addition, unlike the D2 dopamine receptor, the D3 receptor response terminates 15-fold more slowly upon agonist removal. Using D3/D2S chimeric receptors, we demonstrate that D3 receptor tolerance property is mediated by a novel conformational mechanism involving the D3 receptor second cytoplasmic region. The slow response termination rate property requires the third cytoplasmic region and is due to the high affinity of the D3 receptor for ligand as well as its unique G-protein signaling mechanism. PMID- 15121187 TI - GABAergic synapses in hippocampus exocytose aspartate on to NMDA receptors: quantitative immunogold evidence for co-transmission. AB - We previously found evidence for the exocytosis of aspartate from excitatory nerve terminals in hippocampus [J. Neurosci. 18, (1998) 6059]. Here we show, by immunogold electron microscopy in hippocampal slices that aspartate is co localized and co-exocytosed with GABA from synaptic vesicles in nerve endings assumed to be inhibitory on dentate granule cells and CA1 pyramidal cells. By immunogold double labeling cytochemistry in perfusion fixed hippocampus, we further find that GABA-positive terminals forming symmetric synaptic specializations on perikarya of granule and pyramidal cells express NMDA receptors. In addition, NMDA receptors are present at synapses on granule cell bodies that have asymmetric synaptic specializations and contain high levels of GABA. Glutamate levels are low in the described types of GABA-positive nerve terminals, but high in terminals making asymmetric synapses on dendritic spines, whereas aspartate is localized with high levels in all of these types of terminal. We propose that aspartate is exocytotically released not only from glutamatergic terminals, but also from GABAergic terminals to act on NMDA receptors and may have a role in the regulation of synaptic transmission. Under pathological conditions, release of an excitatory transmitter at an inhibitory synapse could contribute to the development of, for example, epilepsy. PMID- 15121188 TI - Transgenic mice overexpressing the full-length neurotrophin receptor trkB exhibit increased activation of the trkB-PLCgamma pathway, reduced anxiety, and facilitated learning. AB - We have investigated the biochemical, physiological, and behavioral properties of transgenic mice overexpressing the full-length neurotrophin receptor trkB (trkB.TK+). The highest trkB.TK+ mRNA overexpression was achieved in the cerebral cortex and hippocampal subfields, both areas also showing strongly increased trkB.TK+ receptor protein expression and phosphorylation. Furthermore, as a result of trkB.TK+ overexpression, partial activation of trkB downstream signaling was observed. Phosphorylation of phospholipaseCgamma-1 was increased but unexpectedly, the expression and phosphorylation levels of signaling molecules Shc and mitogen-activated protein kinase (MAPK) were unaltered. Behavioral studies revealed improved learning and memory in the water maze, contextual fear conditioning, and conditioned taste aversion tests, and reduced anxiety in the elevated plus maze (EPM) and light-dark exploration tests in trkB.TK+ transgenic mice. Electrophysiological studies revealed a reduced long term potentiation (LTP) at the Schaffer collateral-CA1 synapse in trkB.TK+ mice. Altogether, overexpression of the trkB.TK+ receptor postnatally leads to selective activation of trkB signaling pathways and enhanced learning and memory. PMID- 15121190 TI - Reduced GABAergic transmission and number of hippocampal perisomatic inhibitory synapses in juvenile mice deficient in the neural cell adhesion molecule L1. AB - Cell adhesion molecules have been implicated in neural development and hippocampal synaptic plasticity. Here, we investigated the role of the neural cell adhesion molecule L1 in regulation of basal synaptic transmission and plasticity in the CA1 area of the hippocampus of juvenile mice. We show that theta-burst stimulation (TBS) and pairing of low-frequency presynaptic stimulation with depolarization of postsynaptic CA1 pyramidal cells induced similar levels of LTP in L1-deficient and wild-type mice. The basal excitatory synaptic transmission and density of asymmetric excitatory synapses in the stratum radiatum were also normal in L1-deficient mice. Since L1 is expressed not only by principal cells but also by inhibitory interneurons, we recorded inhibitory postsynaptic currents (IPSCs) evoked in CA1 pyramidal cells by minimal stimulation of perisomatic interneurons. L1-deficient mice showed a reduction in the mean amplitude of putative unitary IPSCs, higher values of the coefficient of amplitude variation, higher number of failures in transmitter release, and a reduction in frequency but not amplitude of miniature IPSCs. The use-dependent modulation of inhibitory transmission by paired-pulse or short tetanic stimulation was, however, normal in L1-deficient mice. The physiological abnormalities correlated with a strong reduction in the density of inhibitory active zones, indicating that L1 is involved in establishing inhibitory perisomatic synapses in the hippocampus. PMID- 15121189 TI - Differential expression and dendritic transcript localization of Shank family members: identification of a dendritic targeting element in the 3' untranslated region of Shank1 mRNA. AB - Shank proteins are scaffolding proteins in the postsynaptic density of excitatory synapses in the mammalian brain. In situ hybridization revealed that Shank1/SSTRIP and Shank2/ProSAP1 mRNAs are widely expressed early in postnatal brain development whereas Shank3/ProSAP2 expression increases during postnatal development especially in the cerebellum and thalamus. Shank1 and Shank3 (but not Shank2) mRNAs are present in the molecular layers of the hippocampus, consistent with a dendritic transcript localization. Shank1 and Shank2 transcripts are detectable in the dendritic fields of Purkinje cells, whereas Shank3 mRNA is restricted to cerebellar granule cells. The appearance of dendritic Shank mRNAs in cerebellar Purkinje cells coincides with the onset of dendrite formation. Expression of reporter transcripts in hippocampal neurons identifies a 200 nucleotide dendritic targeting element (DTE) in the Shank1 mRNA. The widespread presence of Shank mRNAs in dendrites suggests a role for local synthesis of Shanks in response to stimuli that induce alterations in synaptic morphology. PMID- 15121191 TI - Collisional cooling enhances the ability to observe non-covalent interactions within the inducible nitric oxide synthase oxygenase domain: dimerization, complexation, and dissociation. AB - The investigation of protein quaternary structure, protein-cofactor, and protein ligand interactions by mass spectrometry is often limited by the fragility of such interactions under experimental conditions. To develop more gentle conditions of perhaps general use, we used as a model for study the oxygenase domain of murine inducible nitric oxide synthase (iNOS), which is homodimeric, binds heme and tetrahydrobiopterin H(4)B cofactors, and the substrate L-arginine. The energetics of the collisions in q2 and in the lens region of the mass spectrometer were manipulated for varying the degree of solvation around the non covalently bound ions. Furthermore, the number of low-energy collisions in the collision cell of the instrument was varied, focusing and dampening the ion beam. Under gentle source collision conditions, and using multiple low-energy collisions in the collision cell of the mass spectrometer, dimers of the iNOS oxygenase domain containing heme, H(4)B, and arginine were observed intact after electrospraying at pH values near neutrality; a mutant of this protein (Trp188 - > Phe) was monomeric and did not bind cofactors. The pH dependence of the iNOS oxygenase domain under acidic conditions was also studied; while heme remained bound to the protein between pH 2.5 and 4.0, the dimeric structure was disrupted. Our findings confirm that non-covalently bound macromolecular complexes are retained and observable using electrospray mass spectrometry under the appropriate experimental conditions. PMID- 15121192 TI - Inter-molecular migration during collisional activation monitored by hydrogen/deuterium exchange FT-ICR tandem mass spectrometry. AB - The difficulty with integrating solution-phase hydrogen/deuterium exchange (HDX) and tandem mass spectrometry is that the energy added to cause fragmentation might promote gas-phase migration of the added deuterium atoms. Here, we compare the solution-phase HDX profiles generated from a- b- and y-type fragment ion series originating from capillary-skimmer dissociation. The isotopic distributions of fragments from the different fragment ion types were used to determine the isotopic state of the amide hydrogen within a specific residue. Even though the same amide hydrogen was examined, the result was different for different fragment ion types. This observation indicates that different fragment series are not equally subjected to inter-molecular migration during collision induced dissociation (CID). We also investigated the gas-phase reactivity of originally undeuterated CID fragments of penta-phenylalanine using gas-phase HDX in an external accumulation hexapole. The incorporation of deuterium into the different fragments was studied as a function of hexapole pressure. It was found that different b- and y-ions from the same peptide had different gas-phase reactivity. However, the a-ions did not display significant gas-phase reactivity. The observed behavior has significant impact on any method that involves comparing the isotopic distributions of different fragment ions. Great care has to be taken in the interpretation of the HDX data using CID to increase the spatial resolution. The isotopic state observed after solution-phase exchange might be more preserved for some CID-fragment types. PMID- 15121193 TI - An isotope labeling strategy for quantifying the degree of phosphorylation at multiple sites in proteins. AB - A procedure for determining the extent of phosphorylation at individual sites of multiply phosphorylated proteins was developed and applied to two polyphosphorylated proteins. The protocol, using simple chemical (Fischer methyl esterification) and enzymatic (phosphatase) modification steps and an accessible isotopic labeling reagent (methyl alcohol-d(4)), is described in detail. Site specific phosphorylation stoichiometries are derived from the comparison of chemically identical but isotopically distinct peptide species analyzed by microspray liquid chromatography-mass spectrometry (microLC-MS) using a Micromass Q-TOF2 mass spectrometer. Ten phosphorylation sites were unambiguously identified in tryptic digests of both proteins, and phosphorylation stoichiometries were determined for eight of the ten sites using the isotope-coded strategy. The extent of phosphorylation was also estimated from the mass spectral peak areas for the phosphorylated and unmodified peptides, and these estimates, when compared with stoichiometries determined using the isotope-coded technique, differed only marginally (within approximately 20%). PMID- 15121194 TI - Deconvolution of isobaric interferences in mass spectra. AB - The concept of isobar deconvolution using the mass domain and signal intensity based domains is described. The intensity domain-based approach employs the reconstruction of the observed isotope pattern from the isolated patterns of the isobaric species. The quantitative information is adjusted with the use of the least squares algorithm. The mass domain-based approach employs signal deconvolution by forming Gaussian components for which the peak width and position can be predicted a priori. The latter method is applicable to medium resolution instruments, such as TOF-MS, while the pattern reconstruction approach is applicable also to low resolution instrumentation, such as quadrupole-based ICP-MS or GC/MS. An example of CH(z)Se (z = 0-4) cluster analysis in dimethyl diselenide mass spectra is given to illustrate the concepts underlying both approaches. PMID- 15121195 TI - Structural determination of neutral O-linked oligosaccharide alditols by negative ion LC-electrospray-MSn. AB - Neutral O-linked oligosaccharides released from the salivary mucin MUC5B were separated and detected by negative ion LC-MS and LC-MS(2). The resolution of the chromatography and the information obtained from collision induced dissociation of detected [M - H](-) ions were usually sufficient to identify the sequence of individual oligosaccharides, illustrated by the fact that 50 different oligosaccharides ranging from disaccharides to nonasaccharides could be assigned from the sample. Fragmentation was shown to yield mostly reducing end sequence fragments (Z(i) and Y(i)), enabling primary sequence assignment. Specific fragmentation pathways or patterns were also detected giving specific linkage information. The reducing end core (Gal/GlcNAcbeta1-3GalNAcol or Gal/GlcNAcbeta1 3(GlcNAcbeta1-6)GalNAcol) could be deduced from the pronounced glycosidic C-3 cleavage and A(i) type cleavages of the reducing end GalNAcol, together with the non reducing end fragment from the loss of a single substituted GalNAcol. Substitution patterns on GlcNAc residues were also found, indicative for C-4 substitution ((0,2)A(i) - H(2)O cleavage) and disubstitution of C-3 and C-4 (Z(i)/Z(i) cleavages). This kind of fragmentation can be used for assigning the mode of chain elongation (Galbeta1-3/4GlcNAcbeta1-) and identification of Lewis type antigens like Lewis a/x and Lewis b/y on O-linked oligosaccharides. In essence, negative ion LC-MS(2) was able to generate extensive data for understanding the overall glycosylation pattern of a sample, especially when only a limited amount of material is available. PMID- 15121196 TI - A G2(+) level investigation of the gas-phase non-identity SN2 reactions of halides with halodimethylamine. AB - The gas-phase non-identity S(N)2 reactions on nitrogen Y(-) + NMe(2)X --> NMe(2)Y + X(-) (Y, X = F, Cl, Br, and I) were evaluated at the G2(+) level. The reactions are exothermic only when the nucleophile is the lighter halide. The complexation enthalpies for complexes Y(-) em leader Me(2)NX are found to correlate with electronegativity of X. Both central and overall barriers can be interpreted with the aid of Marcus equation. Kinetic and thermodynamic investigations predict that the nucleophilicity of X(-) decreases in the order: F(-) > Cl(-) > Br(-) > I(-) and the leaving-group ability increases in the order: F < Cl < Br < I. PMID- 15121197 TI - Compositional analysis of isobutylene/p-methylstyrene copolymers by matrix assisted laser desorption/ionization mass spectrometry. AB - A low molecular weight predominantly polyolefin copolymer of isobutylene and para methylstyrene (IMS) was studied using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Average composition information derived from the spectra was skewed to higher para methylstyrene (pMS) content as compared to that obtained using multiple NMR techniques, and drifted towards lower pMS incorporation at higher oligomer lengths. Although both observations were initially attributed in total to an inability to ionize the isobutylene component, comparison with subsequent field desorption (FD) mass spectrometry results gave similar values to that obtained via MALDI, even though FD ionizes oligomers not detected by MALDI. Instead, the compositional drift observed with MALDI roughly mirrored the mass distribution, and was determined to arise from a mass bias effect in oligomer ionization and detection. Composition with respect to oligomer mass was found to be relatively constant, although similarly higher in pMS content. Comparison of experimental peaks with a Bernoullian statistical model revealed severe overrepresentation of higher pMS composition oligomers with regard to the calculated distribution. This discrepancy is attributed to preferential ionization of oligomers with greater pMS content, and likely results in the observed difference between MALDI and NMR compositions. PMID- 15121198 TI - Cleavage reactions of the complex ions derived from self-complementary deoxydinucleotides and alkali-metal ions using positive ion electrospray ionization with tandem mass spectrometry. AB - The dissociation reactions of the adduct ions derived from the four self complementary deoxydinucleotides, d(ApT), d(TpA), d(CpG), d(GpC), and alkali metal ions were studied in detail by positive ion electrospray ionization multiple-stage mass spectrometry (ESI-MS(n)). For the [M + H](+) ions of the four deoxydinucleotides, elimination of 5'-terminus base or loss of both of 5' terminus base and a deoxyribose were the major dissociation pathway. The ESI MS(n) spectra showed that Li(+), Na(+), and Cs(+) bind to deoxydinucleotides mainly by substituting the H(+) of phosphate group, and these alkali-metal ions preferred to bind to pyrimidine bases rather than purine bases. For a given deoxydinucleotide, the dissociation pathway of [M + K](+) ions differed clearly from that of [M + Li](+), [M + Na](+), and [M + Cs](+) ions. Some interesting and characteristic cleavage reactions were observed in the product-ion spectra of [M + K](+) ions, including direct elimination of deoxyribose and HPO(3) from molecular ions. The fragmentation behavior of the [M + K](+) and [M + W](+) (W = Li, Na, Cs) adduct ions depend upon the sequence of bases, the interaction between alkali-metal ions and nucleobases, and the steric hindrance caused by bases. PMID- 15121199 TI - Fragmentation of protonated ions of peptides containing cysteine, cysteine sulfinic acid, and cysteine sulfonic acid. AB - The oxidation of the sulfhydryl group in cysteine to sulfenic acid, sulfinic acid, and sulfonic acid in proteins is important in a number of enzymatic processes. In this study we examined the fragmentation of four peptides containing cysteine, cysteine sulfinic acid (Cys-SO(2)H), and cysteine sulfonic acid (Cys-SO(3)H) in an ion-trap mass spectrometer. Our results show that the presence of a Cys-SO(2)H in a peptide leads to preferential cleavage of the amide bond at the C-terminal side of the oxidized cysteine residue. The results are important for the determination of the site of the cysteine oxidation and might be useful for the sequencing of cysteine-containing peptides. PMID- 15121200 TI - De novo sequencing, peptide composition analysis, and composition-based sequencing: a new strategy employing accurate mass determination by fourier transform ion cyclotron resonance mass spectrometry. AB - A new strategy is described for the determination of amino acid sequences of unknown peptides. Different from the well-known but often inefficient de novo sequencing approach, the new method is based on a two-step process. In the first step the amino acid composition of an unknown peptide is determined on the basis of accurate mass values of the peptide precursor ion and a small number of accurate fragment ion mass values, and, as in de novo sequencing, without employing protein database information or other pre-information. In the second step the sequence of the found amino acids of the peptide is determined by scoring the agreement between expected and observed fragment ion signals of the permuted sequences. It was found that the new approach is highly efficient if accurate mass values are available and that it easily outstrips common approaches of de novo sequencing being based on lower accuracies and detailed knowledge of fragmentation behavior. Simple permutation and calculation of all possible amino acid sequences, however, is only efficient if the composition is known or if possible compositions are at least reduced to a small list. The latter requires the highest possible instrumental mass accuracy, which is currently provided only by fourier transform ion cyclotron resonance mass spectrometry. The connection between mass accuracy and peptide composition variability is described and an example of peptide compositioning and composition-based sequencing is presented. PMID- 15121201 TI - Using non-covalent complexes to direct the fragmentation of glycosidic bonds in the gas phase. AB - An investigation of the gas phase chemistry of proton bound oligosaccharide (S) ligand (L) non-covalent complexes, [S + H + L](+) has been carried out using electrospray ionization (ESI) and tandem mass spectrometry in a quadrupole ion trap. When subjected to collision-induced dissociation (CID), these [S + H + L](+) complexes undergo a range of reactions that can be broadly classified into three main types: (1) Simple dissociation into the individual monomers; (2) cleavage of the oligosaccharide to form B-type sequence ions; (3) cleavage of the ligand species. The second type of reaction is particularly interesting as it can produce a "ladder series" of [B(x) + L](+) ions via ligand induced oligosaccharide bond cleavage. This novel gas phase reaction greatly simplifies the sequencing of oligosaccharides. Both the oligosaccharide and ligand were found to influence the type of reaction pathway observed, with the "ladder series" of [B(x) + L](+) ions being favored for permethylated oligosaccharides and for bifunctional ligands. Cytosine is a particularly good ligand at facilitating the formation of [B(x) + L](+) ions. Analogies with condensed phase chemistry of sugars is made and a potential mechanism for ligand induced oligosaccharide bond cleavage is proposed. PMID- 15121202 TI - Matrix-assisted laser desorption/ionization tandem mass spectrometry and post source decay fragmentation study of phenylhydrazones of N-linked oligosaccharides from ovalbumin. AB - N-linked oligosaccharides were released from hen ovalbumin by PNGase F and derivatized with phenylhydrazine. They were then examined by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Phenylhydrazones of N glycans under MALDI-tandem mass spectrometry (MS/MS) and post-source decay (PSD) conditions produced relatively similar fragmentation patterns; however, more cross-ring cleavages and fragment ions corresponding to low abundance isomeric structures were detected by MS/MS and not in PSD. Most fragment ions corresponded to glycosidic cleavages with preferential loss of residues from the chitobiose core and the 3-antenna. Sialylated phenylhydrazone-N-glycans, characterized here for the first time in ovalbumin by tandem mass spectrometry, underwent losses of sialic acid residues followed the same fragmentation pathways observed with neutral derivatized glycans. The relative abundances of some fragment ions indicated the linkage position of sialic acid and provided information on the number of residues attached to the 6-antenna. Also, new structures of ovalbumin glycans were observed as part of this study and are reported here. PMID- 15121203 TI - Unexpected products from the reaction of the synthetic cross-linker 3,3' dithiobis(sulfosuccinimidyl propionate), DTSSP with peptides. AB - Synthetic cross-linking reagents, such as 3,3'-dithiobis(sulfosuccinimidyl propionate), DTSSP, can react with sidechains of amino acids that are within close proximity. Identification of cross-linked residues provides insight into the folded structures of proteins. However, analysis of proteolytic digests of proteins cross-linked with commercially available DTSSP is difficult because many ions cannot be attributed to reported reactions of DTSSP. To better understand the reactivity of DTSSP, products from the reaction of DTSSP with several model peptides were analyzed by HPLC electrospray ionization mass spectrometry (ESIMS). Several products not previously reported were identified. Sources for these unexpected products were traced to reaction of DTSSP with contaminant ammonium ions in the buffer, to reaction of contaminants present in the commercial DTSSP reagent, and to reactivity of DTSSP with serine and tyrosine residues. In addition, the collision-induced-dissociation (CID) of peptides modified by DTSSP was investigated. These results showed that certain DTSSP-peptide adducts easily undergo in-source fragmentation to give additional unexpected ions. This study of the reactions of DTSSP with model peptides has revealed the major types of ions that are likely to be found in proteolytic digests of proteins cross-linked with DTSSP, thereby facilitating identification of the cross-linked residues that can provide information about the three-dimensional structures of folded proteins. PMID- 15121205 TI - Review of the 19th Asilomar Conference on Mass Spectrometry: bimolecular interactions: identification and characterization of protein complexes. PMID- 15121204 TI - Glycoprotein profiling by electrospray mass spectrometry. AB - This work compares several different methods of site-specific analysis of glycoproteins using electrospray mass spectrometry. The glycoprotein, oLHalpha (ovine luteinizing hormone, alpha-subunit) was chosen as an appropriate example protein for these studies because of its biological relevance and extreme microheterogeneity. More than 20 unique glycoforms were detected for this glycoprotein at the Asn(56) site of oLHalpha. The carbohydrates present at this site affect receptor binding affinity, so understanding the great variety in the composition of these carbohydrates is important in studying ligand binding interactions. MS data was acquired on a quadrupole ion trap, a triple quadrupole, and a quadrupole time of flight mass spectrometer, and carbohydrate composition at the Asn(56) site of oLHalpha was determined using these instruments. Additionally, neutral loss and precursor ion scanning modes were also used to identify the glycoforms present, and these techniques were compared to the standard MS data. Of the three instruments compared in the study, the qTOF mass spectrometer achieved the lowest sample consumption, but all three instruments were useful in profiling the glycopeptide composition. PMID- 15121206 TI - A statistical method for evaluation quality of medical images: a case study in bit discarding and image compression. AB - Many studies have been performed on quality evaluation for subtle differences in medical images. However, only limited success has been achieved. In this paper, medical images were prior manipulated by denoising, lossy compression and filtering. The Moran statistics is then applied to extract spatial information of images and using Kolmogorov-Smirnov (KS) test to determine whether the manipulated and original images differ significantly. Results show that on average discarding 1-2 bits in T1 and CR images or 2-3 bits in T2 and body CT images are indistinguishable. This method is also applied to a reconstructed MR, body CT image and an electronic SMPTE (Society of Motion Picture and Television Engineer) phantom from lossy image compression software. Compression ratios of 16:1 for a MR image, 8-9:1 for a cropped body CT image, 7:1 and 5:1 for high- and low-resolution regions in electronic phantom is proved undifferentiated from original. The proposed method is useful for complementing the human visual system, to optimize the performance of image compression technique. PMID- 15121207 TI - Development and evaluation of MRI based Bayesian image reconstruction methods for PET. AB - A maximum a posteriori algorithm, which incorporates correlated magnetic resonance images into the processing of positron emission tomography reconstruction with the aim of improving image quality was developed. The line site map from MRI a priori is made up of a modified Markov random field or Canny edge detector with Gaussian smoothing filter. It is used in the MAP algorithm by a weighted line site method. We evaluate and compare the performance of these reconstruction methods. The results show that the Bayesian methods produce reconstructed images with less noise and better spatial resolution than those produced by the maximum likelihood-expectation maximization method. PMID- 15121208 TI - An image analysis approach for automatically re-orienteering CT images for dental implants. AB - In the last decade, computerized tomography (CT) has become the most frequently used imaging modality to obtain a correct pre-operative implant planning. In this work, we present an image analysis and computer vision approach able to identify, from the reconstructed 3D data set, the optimal cutting plane specific to each implant to be planned, in order to obtain the best view of the implant site and to have correct measures. If the patient requires more implants, different cutting planes are automatically identified, and the axial and cross-sectional images can be re-oriented accordingly to each of them. In the paper, we describe the defined algorithms in order to recognize 3D markers (each one aligned with a missed tooth for which an implant has to be planned) in the 3D reconstructed space, and the results in processing real exams, in terms of effectiveness and precision and reproducibility of the measure. PMID- 15121209 TI - A digital reference model of the human bronchial tree. AB - In-vitro preparations of the human lung combined with high-resolution tomography can be used to derive precise models of the human lung. To develop an abstract graph representation, specially adapted image processing algorithms were applied to segment and delineate the bronchi. The graph thus obtained contains topological information about spatial coordinates, connectivities, diameters and branching angles of 1453 bronchi up to the 17th Horsfield order. The graph was analyzed for statistical and fractal properties and was compared with current models. Results indicate a model that exhibits asymmetry and multifractal properties. This newly established reference model is an important step forward in geometrical accuracy of the bronchial tree representation that will improve both analysis of lung images in clinical imaging and the realism of functional simulations. PMID- 15121210 TI - Demonstration of residual perifollicular pigmentation in localized vitiligo--a reverse and novel application of digital epiluminescence dermoscopy. AB - Digital epiluminescence dermoscopy (microscopy) is usually employed to examine melanomas and other pigmented lesions. We report its reverse application in assisting the early diagnosis of a depigmentation condition-localized vitiligo. A pattern of depigmentation with residual reservoirs of perifollicular pigments is clearly visualized. This pattern is not seen in other disorders of depigmentation. Such pattern signifies focally active or repigmenting vitiligo and thus, clearly serves as a useful guide for the cases wherein there exists a doubt about the possible diagnosis. Further studies are warranted to affirm the specificity and applicability of our observations. PMID- 15121211 TI - A tunable incremental factor augmented inverse image alignment method in fundus angiogram registration and mosaicing. AB - A tunable incremental factor is introduced into the inverse compositional image alignment method in registration of fundus blood vessel angiograms under confocal scanning laser ophthalmoscope. The augmented version need less iteration to converge and hardly causes degradation to the registration precision, mainly due to the favorable convexity of the sum of squared differences (SSD) function between the angiograms around the correct registration. And as a compromise between the fidelity of diagnostic information and visual purpose, an adaptive blending strategy based on undecimated discrete sequence wavelet transform is presented in this paper to mosaic fundus angiograms. The effectiveness and efficiency of the tunable incremental factor algorithm is exemplified in the experimental results on clinical indocyanine green angiograms, and the convexity of the SSD function under ideal imaging condition is also illustrated in the paper. PMID- 15121212 TI - Nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase activity in the brain of a cichlid fish, with remarkable findings in the entopeduncular nucleus: a histochemical study. AB - In the entire brain of the African cichlid fish, Haplochromis burtoni, nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity was visualized histochemically in fewer nuclei compared to other teleost fish. Intensively labeled perikarya were found in the ventral hypothalamic area, the nucleus of the medial longitudinal fascicle, the nucleus of the midbrain tegmentum, the nucleus of the lateral longitudinal fascicle, the trigeminal motor nucleus and the octavolateral area. Compared to other NADPH-d labeled nuclei in the brain, we saw an unusual localization of NADPH-d activity in the rostral, dorsal, ventral and caudal part of the entopeduncular nucleus. Additionally, some isolated perikarya of different morphological appearance were found at the levels of the preglomerular nucleus, the diffuse nucleus of the lateral torus and the lateral longitudinal fascicle. A widespread distribution of labeled fibers was identified throughout the brain. The remarkable NADPH-d activity, particularly in the entopeduncular nucleus, differs significantly from the existing data on other teleosts. Taking into account the sensory functions of the entopeduncular nucleus described in other vertebrates, the massive NADPH-d activity in this nucleus may indicate an important role of NADPH-d in the modulation of sensory functions. PMID- 15121213 TI - Cellular distribution of the endothelin system in the human brain. AB - The vasoconstrictor endothelin-1 (ET-1) may also act as a neuropeptide. ET-1 is formed by the catalytic action of endothelin-converting enzyme-1 (ECE-1) on big ET-1 and its cellular actions are mediated via ET(A) and ET(B) receptors. Although localisation of these components in rodent brain has been extensively investigated, no single study has mapped their distribution in human brain. Here we describe the localisation of ET-1 mRNA, ET-1, ECE-1, ET(A) and ET(B) receptors within 24 human brain regions. In situ RT-PCR has previously detected ET-1 mRNA in 22 areas (excluding the post-central gyrus and pineal gland), and ET-1 immunoreactivity was visualised in cells of all regions. Using specific antibodies we have immunolocalised ECE-1 and ET(B) receptors in cells of 24 areas, and ET(A) receptors in nine regions (choroidal epithelial cells, neurones in the diencephalon, hippocampus, amygdaloid, dentate nucleus, Purkinje cells of the cerebellum, flocculo-nodular lobe and vermis). ET-1 mRNA, ET-1, ECE-1 and ET(B) receptors were observed in cortical pyramidal cells, neurones (brainstem, basal nuclei, thalamus, insula and claustrum, limbic region), cells in the anterior pituitary gland; nerve cell processes in the pars nervosa; pinealocytes and choroidal epithelial cells. Only ET-1 mRNA, ET-1, ECE-1, and ET(B) receptors were visualised in cerebral capillary endothelial cells. The presence of ET-1 mRNA, ECE-1 and ET-1 in 22 brain regions confirms ET expression and processing in human brain. The localisation of ET-1 and ET(B) receptors suggests receptor mediated action akin to a neurotransmitter role for ET-1. PMID- 15121214 TI - Differential distribution of MAP1A isoforms in the adult mouse barrel cortex and comparison with the serotonin 5-HT2A receptor. AB - Microtubule-associated protein 1A (MAP1A) is essential during the late differentiation phase of neuronal development. Here, we demonstrated the presence of two MAP1A isoforms with a differential spatial distribution in the adult mouse barrel cortex. Antibody A stained MAP1A in pyramidal and stellate cells, including dendrites that crossed layer IV in the septa between barrels. The other antibody, BW6 recognized a MAP1A isoform that was mainly confined to the barrel hollow and identified smaller caliber dendrites. Previously, an interaction of MAP1A and the serotonin 5-hydroxytryptamine 2A (5-HT(2A)) receptor was shown in the rat cortex. Here, we identified, by double-immunofluorescent labeling, MAP1A isoform and serotonin 5-HT(2A) receptor distribution. MAP1A co-localized mainly with 5-HT(2A) receptor in larger apical dendrites situated in septa. This differential staining of MAP1A and a serotonin receptor in defined barrel compartments may be due to changes in the expression or processing of MAP1A during dendritic transport as a consequence of functional differences in processing of whisker-related sensory input. PMID- 15121215 TI - Histological and immunocytochemical characterization of neurons located in the white matter of the spinal cord of the pigeon. AB - In the spinal cord of birds a considerable number of neuronal somata is located outside the gray matter. Some of these neurons form segmental marginal nuclei, which lie at the border of the spinal cord near the dentate ligament. In lumbosacral segments these marginal nuclei form accessory lobes which bulge into the vertebral canal. These lobes consist in neurons which are embedded into glia derived glycogen cells. Furthermore, there are neurons in the white matter near the accessory lobes and numerous paragriseal cells lying in the lateral and ventral funiculus. Glycogen cells are present both in the lobes and in the glycogen body which fills the lumbosacral spinal rhomboid sinus. Immunoreactivity of glial fibrillary acidic protein, a marker of astrocytes, was used to characterize the surrounding of marginal neurons. Astrocytes were numerous in cervical marginal nuclei but rare in accessory lobes. There is cytological (distribution of Nissl substance) and immunocytochemical evidence (immunoreactivity of medium-sized neurofilament, glutamic acid decorboxylase and glutamatergic AMPA receptor subtype GluR2/3) that neurons of the accessory lobes and the nearby white matter are similar, whereas paragriseal cells are different. PMID- 15121216 TI - Opposite alterations of NPFF1 and NPFF2 neuropeptide FF receptor density in the triple MOR/DOR/KOR-opioid receptor knockout mouse brains. AB - Mice lacking the mu-delta-kappa-opioid receptor (MOR/DOR/KOR) genes and their corresponding wild-type littermates have been used to quantify NPFF(1) and NPFF(2) (neuropeptide FF) receptors by in vitro autoradiography in the central nervous tissues. Adjacent coronal sections were labelled with [125I]YVP ([125I]YVPNLPQRF-NH(2)) and [125I]EYF ([125I]EYWSLAAPQRF-NH(2)) as specific radioligands for NPFF(1) and NPFF(2) receptors, respectively. NPFF(2) receptors are predominantly expressed in both genotypes, but their density increases significantly in non cortical regions of mutant mice: 64% in the amygdaloid area, 89, 308, 1214 and 49% in the nucleus of the vertical limb of the diagonal band, substantia nigra, the vestibular nucleus and the spinal cord, respectively. In contrast, the density of the NPFF(1) subtype is lower than NPFF(2) in both genotypes and significantly decreased in some brain areas of mutant mice: -99, 90 and -90% in the nucleus of the vertical limb of the diagonal band, substantia nigra and the spinal cord, respectively. This study shows that mice lacking opioid receptors have brain region-dependent increases (NPFF(2)) and decreases (NPFF(1)) in NPFF receptors densities and suggests a different functional participation of each NPFF receptor subtype in the actions of opioids. PMID- 15121217 TI - The effect of oral 5-HTP administration on 5-HTP and 5-HT immunoreactivity in monoaminergic brain regions of rats. AB - 5-Hydroxytryptophan (5-HTP), which is the rate-limiting precursor in serotonin (5 hydroxytryptamine (5-HT)) biosynthesis, is used as an oral supplement to enhance serotonin levels in humans. To evaluate its effects on serotonin levels and localization, 5-hydroxytryptophan was administered to Sprague-Dawley rats either orally or via intraperitoneal injection. 5-Hydroxytryptophan-immunoreactivity was co-localized with serotonin-immunoreactivity in the serotonergic dorsal raphe nucleus of control animals and this was not changed in animals given 5 hydroxytryptophan. Oral 5-HTP administration increased the intensity of both 5 HTP and serotonin immunoreactivity in raphe neurons. However, 5-HTP treatment also caused ectopic 5-hydroxytryptophan-immunoreactivity and serotonin immunoreactivity in normally dopaminergic neurons of the substantia nigra par compacta. Serotonin-immunoreactivity was confined to neurons that also displayed amino acid decarboxylase immunoreactivity, but in a small percentage of substantia nigra neurons, serotonin immunoreactivity was not co-localized with tyrosine hydroxylase-immunoreactivity. The intensity of the immunoreactivity to serotonin and 5-hydroxytryptophan in the substantia nigra was maximal within 2h of 5-hydroxytryptophan administration and returned to control levels by 24h. This time course mirrored changes in HPLC measurements of 5-hydroxytryptophan, serotonin, and the metabolite 5-hydroxyindoleacetic acid (5-HIAA) in the urine. 5 Hydroxytryptophan administration did not cause ectopic appearance of either serotonin or 5-hydroxytryptophan in the noradrenergic locus coeruleus. These results suggest that a single oral dose of 5-HTP increases the 5-HTP and serotonin content of serotonergic neurons and causes the transient ectopic appearance of serotonin in some normally non-serotonergic neurons. PMID- 15121218 TI - The role of chemotherapy for advanced oro and hypopharyngeal cancer. AB - Although important progress continuous to be made in the treatment of oro and hypopharyngeal cancer, the 5-year survival rate for all this disease has remained at less than 30% for the past 30 years. In the early 1980s, chemotherapy was introduced with high expectation of reducing in the incidence of distant metastases and increasing the possibility of local control. This article explores the use of chemotherapy in the treatment of advanced pharyngeal cancer. Thus, the efficacy of chemotherapy are reviewed and treatment options for advanced pharyngeal cancer are made. When advanced carcinoma is still localized, function preserving surgery is performed. In these cases, the possibility of instituting adjuvant chemotherapy with an active treatment regimen may be taken into account depending on the condition of the patient and the tumor. Patients with surgically resectable tumors are given 1-2 cycles of induction chemotherapy. Cases who respond to the induction chemotherapy are subsequently given concurrent chemoradiotherapy. Residual lymph nodes in the neck are removed surgically. The cases who do not respond to the induction chemotherapy are treated with radical surgery. Patients with unresectable carcinoma are given concurrent chemoradiotherapy because local treatment should be performed in such patients as early as possible. In principle, concurrent regimens should be supplemented with adjuvant chemotherapy in all cases. This is particularly required for those with advanced N-stage patients. PMID- 15121219 TI - Semilongitudinal and axial CT planes in assessing cochlear patency in cochlear implant candidates. AB - OBJECTIVE: To investigate how cochlear patency as seen on computed tomography (CT), using axial plus semilongitudinal planes, is correlated with findings at surgery in cochlear implant patients. METHODS: Pre-operative CT scans of 45 patients were reviewed by three, independent observers. They classified the cochlear patency and recorded the location of any suspected decreased patency. The results were compared with the findings noted during surgery. RESULTS: In nine patients a decreased cochlear patency was found at surgery. The sensitivity and specificity of CT assessment were, respectively, 56-33-11% and 100-86-94%. The interobserver reproducibility is reflected in a mean kappa of 0.46. The sensitivity increased when only patients suffering from post-meningitic deafness were considered. CONCLUSION: Our study suggests that CT scans can be useful in assessing cochlear patency, especially in patients with post-meningitic deafness. This good performance might be explained by the combined use of scans in semilongitudinal and axial planes. PMID- 15121220 TI - Correlation of CT analysis and audiometry in Japanese otosclerosis. AB - OBJECTIVE: To determine the extent of audiometric correlation with CT findings. METHODS: Forty-four patients (82 ears) with surgically confirmed otosclerosis underwent preoperative CT examination. Based on the computed tomography (CT) findings, the ears were classified into five groups as follows: Group A, the group with no pathological CT findings; Group B1, the group with demineralization localized in the region of the fissula antefenestram; Group B2, the group with demineralization extending towards the cochleariform process from the anterior region of the oval window; Group B3, the group with extensive demineralization surrounding the cochlea; and Group C, the group with thick anterior and posterior calcified plaques. RESULTS: There were 32 ears (39.0%) in Group A, 21 ears (25.6%) in Group B1, 16 ears (19.5%) in Group B2, 7 ears (8.5%) in Group B3, and 6 ears (7.3%) in Group C. The mean bone conduction levels were greater in the order of the extent of demineralization: Groups A, B1-B3 suggesting positive relationship between the cochlear function and the degree of labyrhinthine otosclerosis. CONCLUSION: A good correlation between the preoperative CT findings and audiometry findings suggests that CT with a slice intervals between 0.5 and 1mm could provide useful informations in assuming the extent of otosclerosis in the inner ear. PMID- 15121221 TI - Indications and results of cauterization by endoscopic approach of the sphenopalatine artery in severe posterior epistaxis. AB - OBJECTIVES: The authors present a retrospective study concerning cauterization by endoscopic approach of the sphenopalatine artery in the treatment of severe posterior epistaxis. The purpose is to evaluate the effectiveness of these therapeutics, their complications, to determine their indications and their contributions to the therapeutic arsenal of the treatment of severe epistaxis. PATIENTS AND METHOD: This study concerns 10 patients, which have been operated in the department of ORL head and neck surgery of Strasbourg and treated over a 23 month period (from January 2001 to November 2002), for clinically labelled posterior origin epistaxis. A preliminary treatment, by anterior and posterior tamponage, using a Brighton Epistaxis Balloon was carried out (n = 8); the use of an associated ligature of the ethmoidal arteries was sometimes necessary during operative time (n = 4). RESULTS: Hemorrhagic control with absence of recurrence was the rule (n = 9). A complementary interventional radiology embolization due to failure was necessary in one case. The average duration of post-surgical hospitalization was of 2.1 days. No post-surgical complications were noted. CONCLUSION: Cauterization by endonasal approach of the sphenopalatine artery in severe posterior epistaxis is a reliable intervention. Its role is clearly defined in our department, after failure of traditional treatments by packing methods. PMID- 15121222 TI - Measurement of quality of life during different clinical phases of Japanese cedar pollinosis. AB - OBJECTIVE: To assess Quality of Life (QOL) scores in patients with Japanese cedar (Cryptomeria japonica, CJ) pollinosis. METHODS: QOL was measured intraseasonally and extraseasonally in patients (n = 69) using the Medical Outcomes Study Short Form Health Survey, and compared with healthy control subjects (n = 50). The differences in intraseasonal QOL scores between short and long disease duration groups were studied. The relation of intraseasonal QOL scores to nose symptoms was evaluated using Spearman's rank correlation. RESULTS: Significant decrease was observed in two of eight QOL items in intraseasonal measurements of patients. These QOL impairments improved extraseasonally. Scores for emotional problems in patients with long disease duration were less impaired than those with short disease duration. QOL scores in patients correlated poorly with the severity of nasal symptom. CONCLUSIONS: This is the first study to report QOL impairment in CJ pollinosis. Based on the findings long disease duration may improve emotional problems, and the correlation between disease severity and QOL is low. PMID- 15121223 TI - Taste function in Sjogren's syndrome patients with special reference to clinical tests. AB - OBJECTIVE: We analyzed the correlation between taste functions and clinical features in patients with Sjogren's syndrome (SS). METHODS: Sixty-three patients were divided into two groups: group A consisting of patients with primary or secondary SS of the European classification, and group B consisting of non-SS complaining of dry eye and mouth. RESULTS: Eighteen percent of the group A patients and 11% of the group B patients complained of taste disorders. Electrogustometric examination detected taste disorders in 27% of the group A patients and 38% of the group B patients, while the filter paper disc method showed taste disorders in 30% of group A and 40% of group B. In group A, unstimulated and stimulated salivary flows were correlated with the ability to discriminate taste analyzed by the filter paper disc method, but not with the taste electrogustometric threshold. Serum zinc concentration was not significantly correlated with taste disorders in either group. CONCLUSION: Our findings showed that SS patients presented taste disorders less frequently than previously reported, and we speculated that taste disorder found in SS patients were due to the salivary flow deficiency that might prevent substances from reaching the taste buds. PMID- 15121224 TI - A dendritic cell based hybrid cell vaccine generated by electrofusion for immunotherapy strategies in HNSCC. AB - OBJECTIVE: Hybrid cells generated from dendritic cells (DC) and tumor cells provide tumor-associated antigens (TAA) in a polyvalent mode and therefore they have aroused interest in cancer immunotherapy. The present study was designed to investigate the hybrid cell generation and optimize its implementation for a TAA target treatment of head and neck squamous cell carcinoma (HNSCC). METHODS: Hybrid cells from mature DC and laryngeal carcinoma cell line UTSCC-19A were generated by electrofusion. Fusion efficiency and viability were determined by flow cytometry, light and fluorescence microscopy analyses. RESULTS: The gradual electrofusion process constituted real human tumor and dendritic cell hybrids characterized by polynuclear cells and double staining as a result of overlay of red (HLA-DR:R-PE) and green (HEA:FITC) fluorescence. Furthermore, analyses have proven viability of fusion results, and factors influencing fusion yield were determined. CONCLUSION: Physical fusion of mature dendritic cells with laryngeal carcinoma cells provides a dendritic cell based hybrid cell vaccine as a quantitative prerequisite for anti-cancer vaccination. Specific cytotoxic T lymphocytes need to be induced before hybrid cell application in clinical studies. PMID- 15121225 TI - Hypertrophic chronic pachymeningitis associated with chronic otitis media and mastoiditis. AB - We describe the case history of a 70-year-old female patient presenting with bilateral hearing disturbance, facial paralysis, and vertigo. Radiological tests of temporal bone revealed soft tissue in the mastoid and tympanic cavities, and T1 weighted MRI revealed prominent Gd enhancement of the middle skull basal meninges. Middle ear inflammation appeared to induce pachymeningitis and to exacerbate associated symptoms, leading to a decline in the patient's overall condition. Bilateral mastoidectomies were effective in improving her general condition. Her hearing improved only on the right side because ossiculoplasty was performed only on that side. Her facial movement progressively improved and pachymeningitis diminished over time. We speculate that removal of the infectious granulation within the middle ears and mastoids ameliorated the acute inflammation. The etiology remains unknown in this case. PMID- 15121226 TI - Melanin pigmented oncocytic metaplasia of the nasopharynx. AB - A case of melanin pigmented oncocytic metaplasia is reported. A 62-year-old man presented with a history of discomfort of the ear of a 2 weeks' duration. Nasoscopic examination revealed a black nodule of about 5mm at the left Eustachian opening, and several black spots were discovered around the bilateral torus tubarius. The nodule was biopsied to determine the histology. Microscopically, there were oncocytic cells with abundant pigmented granules showing glandular pattern. Such a pigmented variant of benign oncocytic lesion is very rare. PMID- 15121227 TI - Subglottic leiomyosarcoma of the larynx; a case report. AB - Leiomyosarcoma of the larynx is very rare. We herein report an especially rare case of leiomyosarcoma originating in the subglottic mucosa. The majority of cases reported in the literature indicate that laryngeal leiomyosarcomas originate in the supraglottic or glottic area. Our patient underwent simple total laryngectomy and is free of disease 5 years after the operation. The literature is reviewed regarding the choice of treatment and the distribution of the reported sites of origin of laryngeal leiomyosarcoma. PMID- 15121228 TI - Oropharyngeal dysphagia in a case of Huntington's disease. AB - The aim of this study is to analyze dysphagic symptoms of a patient with Huntington's disease (HD) who had having difficulty in swallowing. The patient was a 66-year-old female with HD. Inspection of self-feeding at bedside and videofluorographic swallowing assessment were performed. The features at self feeding were the tendency of rapid eating, inability for smooth transportation of food to oral cavity, weak lip closure, which resulted in falling of food and eating it again. The videofluorography indicated clumsy tongue movement and postural instability by chorea which caused discoordination between oral and pharyngeal stage. Those ended in spill of liquid to the pharynx and retention of bolus in the oral cavity and vallecula, and aspiration did not occur. Pudding was carefully chewed because of the patient's alertness to the examination. The cognitive disturbance and choreic movement caused dysphagia at the preparatory and oral stages, and chorea also produced the discoordication between the oral and pharyngeal stage. The change of the shape of cups and stable posture were advised to lessen the chance of her aspiration. PMID- 15121229 TI - Spindle cell carcinoma of the larynx. AB - Four rare cases of spindle cell carcinoma of the larynx are reported and the histo-pathological and immuno-histochemical findings are described. The spindle cell carcinoma involved the right vocal cord of an 88-year-old and an 86-year-old male patients and the anterior commissure of a 76-year-old and a 68-year-old male patients. Two patients underwent total laryngectomy, one patient extirpation under laryngomicrosurgery and one radiation therapy. All were well controlled thereafter and there was no recurrence. Surgery seemed to be the most satisfactory therapeutic approach and radiation therapy was effective in a patient with relatively small tumor. The histo-pathological and immuno histochemical analysis in one patient revealed a sarcoma-like component of the carcinoma and the findings suggested that this was the result of a metaplastic change in the mesenchyme without inflammatory changes. PMID- 15121230 TI - A case of mediastinal goiter. AB - A case of mediastinal goiter in a 51-year-old female is reported. She demonstrated an abnormal shadow on chest X-ray but there were no clinical symptoms. Tumors were recognized in the isthmus of the thyroid and mediastinum on CT scan and MRI. These two tumors were resected via standard neck skin incision and sternotomy. At surgery, we found no communication between the two tumors. Following Rives' classification, the mediastinal tumor in this case was diagnosed as aberrant mediastinal goiter. Pathological diagnoses of both tumors were adenomatous goiter. PMID- 15121231 TI - A rare salivary gland neoplasm: multiple canalicular adenoma; A case report. AB - The canalicular adenoma is an uncommon, benign salivary gland tumour that most frequently occurs in the upper lip. Although the incidence of multifocal epithelial tumours of the minor salivary is very low, canalicular adenoma sometimes present as a multifocal lesion. We present a case of multifocal canalicular adenomas of upper lip in a woman aged 68 years and discuss their features, emphasising diagnosis, clinical behaviour, treatment, histological and immunohistochemical aspects. PMID- 15121232 TI - Two cases of non-Hodgkin's lymphoma in the accessory parotid gland. AB - Primary malignant lymphomas in the salivary glands are relatively rare and tumors of the accessory parotid gland comprise only 1% of parotid tumors. We present two cases with a painless swelling of the cheek region. In both cases histological diagnoses of primary non-Hodgkin's lymphoma were made following complete excision of the accessory parotid gland tumor. PMID- 15121233 TI - Perfusion- and BOLD-based fMRI in the study of a human pathological model for task-related flow reductions. AB - In the present work, an arteriovenous malformation was taken as a pathological model for studying task-related flow decreases during a motor task. Combined Blood Oxygen Level Dependent (BOLD)-perfusion experiments were applied in order to evaluate the relative sensitivity of these techniques to task-related reductions in cerebral blood flow (CBF). Results shows that, by matching the sensitivity of the methods (which exhibit a different contrast-to-noise ratio) in the primary motor cortex, the spatial extent of the regions of decreased perfusion signal is larger than those of the BOLD signal reduction. The above finding suggests that perfusion imaging, that already represents a gold standard method in the detection of vascular phenomena, may estimate task-related flow decreases in a functional time-series better than BOLD. PMID- 15121234 TI - Distinct regional distributions of NK1 and NK3 neurokinin receptor immunoreactivity in rat brainstem gustatory centers. AB - Tachykinins and their receptors are present in gustatory centers, but little is known about tachykinin function in gustation. In this study, immunohistochemical localization of substance P and two centrally prevalent neurokinin receptors, NK1 and NK3, was carried out in the rostral nucleus of the solitary tract and the caudal parabrachial nucleus to evaluate regional receptor/ligand correspondences. All three proteins showed regional variations in labeling density that correlated with distinct sites in gustatory centers. In the rostral nucleus of the solitary tract, the relative densities of substance P and NK1 receptors varied in parallel across subnuclei, with both being moderate to dense in the dorsocentral, chemoresponsive zone. NK3 receptors had a distinct distribution in the caudal half of this zone, suggesting a unique role in processing taste input from the posterior tongue. In the caudal parabrachial nucleus, substance P and NK1 receptor immunoreactivities were dense in the pontine taste area, while NK3 receptor labeling was sparse. The external medial subnucleus had substantial NK3 receptor and substance P labeling, but little NK1 receptor immunoreactivity. These findings suggest that distinct tachykinin ligand/neurokinin receptor combinations may be important in local processing of information within brainstem gustatory centers. PMID- 15121235 TI - The evolution of dysgraphia in Alzheimer's disease. AB - Evidence from recent studies suggests that writing may be an aspect of cognition capable of identifying impairments specific to patients with Alzheimer's Disease (AD). The precise nature and progression of the writing disorder, however, remains unclear. The current study assessed the central and peripheral aspects of writing among a sample of minimal, mild and moderate AD patients and a group of healthy elderly controls on a narrative description task. Comparisons of the two groups indicated that AD patients suffer from a primary impairment at the semantic level. Even those in the minimal stages of the disease could be differentiated from controls on measures of word finding and information conveyed. This semantic impairment was coupled with a secondary milder impairment in phonological processing. The prevalence of phonological errors increased, but no shift in error type (plausible/implausible) was identified as the disease progressed. In addition to the central impairments, patients evinced damage at the peripheral level. In the more severe stages, patients experienced more problems with letter formation and stroke placement and tended to rely upon the more simplistic writing form of print. The writing impairment in AD is multi componential in nature and follows the pattern of cortical deterioration reported in the brains of AD patients. PMID- 15121236 TI - Altered localization of choline transporter sites in the mouse hippocampus after prenatal heroin exposure. AB - Prenatal heroin exposure disrupts hippocampal cholinergic synaptic function and related behaviors. Biochemical studies indicate an increase in the number of presynaptic high-affinity choline transporter (HACT) sites, as assessed by [3H]hemicholinium-3 (HC-3) binding. The present study was designed to assess whether this effect involves global upregulation of the transporter, or whether disruption occurs with a specific tempero-spatial distribution. Pregnant mice were given 10mg/kg per day of heroin subcutaneously on gestational days (GD) 9 18. Autoradiographic distribution of HC-3 binding sites was evaluated in the hippocampus of the offspring at postnatal days 15, 25, and 53. These results, suggestive of hippocampal "miswiring," are likely to explain the net impairment of cholinergic synaptic function after prenatal heroin exposure, despite the simultaneous upregulation of both presynaptic cholinergic activity and postsynaptic receptors. Understanding the subregional selectivity of hippocampal defects can lead to the development of strategies that may potentially enable therapeutic interventions to offset or reverse the neurobehavioral defects. PMID- 15121237 TI - Differential messenger RNA expression of complexins in mouse brain. AB - Complexins (CPLXs) are small isomeric proteins that bind to the soluble NSF attachment protein receptor (SNARE) complex and modulate neurotransmitter release. Two isoforms of CPLX exist in the brain, CPLXI and CPLXII. These are differentially distributed in the cortex and cerebellum, with CPLXI found in axosomatic terminals and CPLXII in axodendritic terminals. Since in cortex and cerebellum axosomatic terminals are inhibitory and axodendritic terminals are excitatory, it has been assumed that CPLXI modulates inhibitory and CPLXII modulates excitatory transmitter release. Here we used in situ hybridisation to study the mRNA distribution of CPLXI and CPLXII in mouse brain. We show that while CPLXs are expressed in distinct cell populations, they do not segregate with either particular neurotransmitters, or different classes of transmitter action. For example, while CPLXII is the dominant isoform in the output (glutamatergic excitatory) neurons of the cortex, it is also the dominant isoform in medium spiny (GABAergic inhibitory) neurons of the striatum. We suggest that the functional role of CPLXs depends not only on the identity of the neurotransmitter, but also upon the circuitry connecting the neurons in which they are expressed. Thus, the predominant expression of CPLXII in neurons of the basal ganglia and cortex suggests a role in cognition, emotional behaviour and control of voluntary movement, while the pattern of CPLXI expression suggests a primary role in motor learning programs and sensory processing. PMID- 15121238 TI - Regional and progressive changes in brain expression of complexin II in a mouse transgenic for the Huntington's disease mutation. AB - Changes in mRNA expression of soluble NSF-attachment protein receptors (SNAREs) and SNARE-associated proteins have been shown to occur in a number of disorders such as schizophrenia, Alzheimer's disease and Parkinson's disease. We have shown previously that there is a decrease in protein levels of the SNARE-associated protein, complexin II (CPLXII) in Huntington's disease brain and in the R6/2 mouse model of Huntington's disease. In the current study, we used quantitative in situ hybridisation to examine mRNA expression of SNAREs (25 kDa synaptosome associated protein (SNAP-25), syntaxin-1A and synaptobrevin-2) and SNARE associated proteins (alpha-SNAP, CPLXI and CPLXII) in brain of R6/2 mice and their wild type littermates between 3 and 15 weeks of age. We found an early and progressive decrease of CPLXII expression in R6/2 mice brains. In contrast, no changes in SNARE expression were seen in R6/2 brains compared with wild type brain. Further, while decreased expression of alpha-SNAP and CPLXI was seen, this was not until 15 weeks of age and even then the changes were small. We suggest that downregulation of expression of mRNA encoding SNARE-associated proteins, first CPLXII and later CPLXI and alpha-SNAP, contributes to the progressive neuropathology of the R6/2 mouse model of Huntington's disease. PMID- 15121239 TI - Gene expression of OGFr in the developing and adult rat brain and cerebellum. AB - The native opioid peptide, [Met5]-enkephalin (termed opioid growth factor (OGF)), is a tonically active negative growth factor targeted to cell proliferation in the developing nervous system. OGF action is mediated by the OGF receptor (OGFr). The present study investigates gene expression of OGFr in the developing and adult brain and cerebellum of the rat using Northern blot analysis and normalization to GAPDH. OGFr was detected in whole brain at embryonic day 20 and birth, and was at least twofold greater than neonatal levels during the first week of life. From postnatal day 15 onwards to adulthood, levels of OGFr mRNA in the whole brain were detectable but less than those at birth. OGFr mRNA in cerebellum was found on embryonic day 20, and remained relatively constant until postnatal day 12 when a sharp increase was recorded. In the third week of life and continuing into adulthood, cerebellar OGFr mRNA was detected at levels comparable to those in postnatal week 1. These results show that message for OGFr is developmentally regulated prior to and after birth, is ubiquitously expressed during development, and is present in the adult brain and cerebellum even though OGF receptor binding is not recorded. PMID- 15121240 TI - Temporal and regional expression of Fos-related proteins in response to ischemic injury. AB - The AP-1 transcription factor family has been widely studied in the response to ischemic brain injury. The data to date have demonstrated a complex involvement that depends on stimulus, subunit composition and brain region. One member in particular, the Fos-related antigen FRA-2, has demonstrated a potential for controlling neuroprotective gene expression. This study characterized the temporal and regional expression of a variety of proteins following ischemic injury induced by occlusion of the middle cerebral artery in rats. The results demonstrated upregulation of both c-Fos and FRA-2 in penumbral regions that preceded upregulation of the classic injury-associated proteins expressed by astrocytes and microglia and, in the case of FRA-2, appeared to correlate with neuronal survival. A further, previously undescribed, expression of FRA-2 in endothelial cells of the core ischemic region was also demonstrated. PMID- 15121241 TI - Effect of central administration of carnosine and its constituents on behaviors in chicks. AB - Even though their contents in the brain are high, the function of brain carnosine and its constituents has not been clarified. Both carnosine and anserine inhibited food intake in a dose dependent fashion when injected intracerebroventricularly. The constituents of carnosine, beta-alanine (beta-Ala) and l-histidine (His), also inhibited food intake, but their effects were weaker than carnosine itself. Co-administration with beta-Ala and His inhibited food intake similar to carnosine, but also altered other behaviors. Injection of carnosine induced hyperactivity and increased plasma corticosterone level, whereas beta-Ala plus His induced hypoactivity manifested as sleep-like behavior. This later effect seemed to be derived from beta-Ala, not His. These results suggest that central carnosine may act in the brain of chicks to regulate brain function and/or behavior in a manner different from its constituents. PMID- 15121242 TI - Rapid decline in motor symptoms in HD neural transplant patients prior to surgery. PMID- 15121243 TI - Contrast echocardiography: new agents. AB - In this report, we review the history, rationale, current status and future directions of contrast agents in echocardiography. First, we discuss the historic development of contrast agents through a review of important physical principles of microbubbles in ultrasonography. Second, we identify attributes of an ideal contrast agent and review those that are currently available or in the "pipeline" for clinical use. Third, we review indications for contrast echocardiography, including endocardial border detection, perfusion quantification and reperfusion assessment, and validate these observations by comparisons with other imaging modalities. Then, we briefly review different methodologies of performing a contrast study, including interrupted, real-time and a hybrid modality. Finally, we identify novel future applications of the newest contrast agents. These newer concepts in contrast echocardiography should form a foundation for nearly limitless application of echocardiography in improved anatomical assessment, perfusion imaging and even special applications, such as detection of vascular inflammation and site-specific drug delivery. PMID- 15121244 TI - Power Doppler findings in plantar fasciitis. AB - The study was performed to characterize the power Doppler ultrasonographic (PDU) findings in plantar fasciitis using a 7.5 MHz linear transducer. Both feet of 20 patients who had a clinical and ultrasound (US) diagnosis of unilateral plantar fasciitis were evaluated with PDU. The pain level was assessed with a visual analogue scale (VAS). A total of 20 healthy volunteers were evaluated as a control group. Moderate or marked hyperemia was found in PDU in the plantar fascia and the surrounding soft tissue along the first cm distally from the insertion in 8 (40%) of the 20 symptomatic heels and in 1 patient (5%) on the asymptomatic side. Moderate or marked hyperemia was associated with a history of less than 6 months and high pain levels. The difference between both groups was significant (p < 0.05). PDU improves the value of US as a noninvasive technique for the diagnosis of plantar fasciitis, providing additional information on local hyperemia. PMID- 15121245 TI - Accuracy of sonographic vascular features in differentiating different causes of cervical lymphadenopathy. AB - This study was undertaken to evaluate the accuracy of the power Doppler sonographic features in differentiating cervical lymphadenopathy in different diseases. We reviewed power Doppler sonograms of 270 patients with palpable neck nodes (metastases n = 101, lymphoma n = 21, tuberculosis n = 76, reactive n = 72). Confirmed diagnosis was either made by fine-needle aspiration cytology on the largest node or by excision biopsy. In each patient, the largest node was included in the study. The node was evaluated using the vascular pattern, displacement of vascularity and vascular resistance (resistance index, RI, and pulsatility index, PI). Individual groups were compared and the optimum vascular feature in the differential diagnosis was determined. Vascular pattern was more useful in differentiating reactive nodes from malignant nodes, with a sensitivity of 88% for metastases and 67% for lymphoma, and a specificity of 100%. RI with a cut-off value of 0.8 was more accurate in distinguishing metastases (RI > 0.8) from lymphoma (RI < 0.8), with an accuracy of 65% and 75%, respectively. Displacement of vascularity was helpful to differentiate tuberculous nodes (accuracy: 67%) from reactive and lymphomatous nodes (accuracy: 100% and 95%, respectively), whereas PI with a cut-off of 1.5 helped the differentiation between tuberculosis (PI < 1.5) and metastases (PI > 1.5), with an accuracy of 77% in both diseases. When appropriate criteria are used, power Doppler sonography is a valuable adjunct in the sonographic evaluation of cervical lymphadenopathy. PMID- 15121246 TI - Grey-scale and power Doppler sonography of unusual cervical lymphadenopathy. AB - This study was undertaken to document the grey-scale and power Doppler sonographic features of cervical lymphadenopathy in Kikuchi's disease (histiocytic necrotising lymphadenitis), Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy), Sjogren's syndrome and systemic lupus erythematosus (SLE), which have not been reported in the literature. A retrospective review of the grey-scale and power Doppler sonograms of the cervical lymph nodes in nine patients was conducted (Kikuchi's disease, n = 3; Rosai-Dorfman disease, n = 1; Sjogren's syndrome, n = 1; SLE, n = 4). Lymph nodes were proven to be pathologic by fine-needle aspiration cytology (FNAC). On grey scale ultrasound (US), lymph nodes were assessed by their distribution, size, shape, echogenicity and internal architecture. The vascular pattern of the lymph nodes was assessed with power Doppler sonography. US features of the lymph nodes were compared to those of metastatic and reactive nodes. In Kikuchi's disease, Rosai-Dorfman disease, Sjogren's syndrome and SLE, the distribution of lymph nodes is similar to that of reactive nodes. Most of the lymph nodes are enlarged with a maximum transverse diameter greater than or equal to 10 mm (83.3 to 100%). In Kikuchi's disease, lymph nodes have grey-scale and Doppler appearances similar to reactive nodes. However, lymph nodes in Rosai-Dorfman disease, Sjogren's syndrome and SLE show similar grey-scale and Doppler features to metastatic nodes. There is no specific US feature to characterise lymphadenopathy from these four miscellaneous causes. Definitive diagnosis should still be based on cytology and histology, and US can help in guiding FNAC for a more accurate cytologic examination. PMID- 15121247 TI - Quantitative ultrasound at hand phalanges in adults with end-stage renal failure. AB - The aim of this study was to assess bone status in 220 subjects with end-stage renal failure (ESRF) (146 men, mean age 53.0 +/- 13.9 years and 74 women, mean age 48.1 +/- 14.3 years). The duration of hemodialysis (durHD) and duration of renal insufficiency (durRI) were, in men, 2.6 +/- 3.8 years and 7.7 +/- 8.0 years, and, in women, 2.8 +/- 3.4 years and 9.1 +/- 7.6 years, respectively. ESRF was caused by the following reasons: chronic glomerulonephritis in 92 patients, diabetes in 52, chronic pyelonephritis in 37, polycystic kidney disease in 19, amyloidosis in 5, hypertension in 4 and unknown cause in 11. The control group consisted of 1615 normal healthy subjects (1216 women, mean age 48.1 +/- 12.1 years and 399 men, mean age 52.9 +/- 14.8 years). Mean age did not differ between patients and controls. Skeletal status was evaluated by quantitative ultrasound (US) measurements at the hand phalanges using DBM 1200 (IGEA, Italy) which measures amplitude-dependent speed of sound (Ad-SoS, m/s). The mean value of Ad SoS in male patients was 1981 +/- 88 m/s, T-score -l2.03 +/- 1.26, Z-score -0.53 +/- 1.7 and, in female patients, 1967 +/- 96 m/s, -2.23 +/- 1.37, -1.41 +/- 1.56, respectively. Respective values in male controls were 2008 +/- 81 m/s, -1.66 +/- 1.16, -0.01 +/- 0.98 and, in female controls, 2026 +/- 81 m/s, -1.4 +/- 1.15, 0.74 +/- 0.86, and were significantly higher than in male (p < 0.001) and female (p < 0.0000001) patients. A correlation analysis of Ad-SoS with durHD and durRI showed that only in males did both factors significantly influence parameters measured (r = -0.26, p < 0.01). Multiple stepwise regression analysis of Ad-SoS on age, durHD, durRI, weight and height was possible to perform only in males and the following equation was established: Ad-SoS = 2545 m/s - 3.09 x age (years) - 5.68 x durHD (years) - 2.15 x height (cm) - 0.99 x durRI (years), p < 0.000001, r = 0.55, SEE = 69.6. Concluding, in subjects with ESRF treated with hemodialysis, skeletal status assessed with the use of quantitative US was affected. PMID- 15121248 TI - The effects of extracorporeal shock-wave therapy on the ultrasonographic and histologic appearance of collagenase-induced equine forelimb suspensory ligament desmitis. AB - Extracorporeal shock-wave therapy (ESWT) may stimulate healing of desmitis in multiple species. The objective of this study was to evaluate the ultrasonographic and histologic appearance of collagenase-induced suspensory ligament (SUL) desmitis in untreated ligaments and ligaments treated with ESWT in horses. Four mature horses had SUL desmitis induced in both forelimbs. Beginning 3 weeks after induction of the lesions, one ligament per horse was treated 3 times at 3-week intervals with ESWT. The percent lesion, echogenicity and fiber alignment scores of the SULs were evaluated at 3-week intervals for a total of 15 weeks. At the completion of the study, the horses were euthanized and the SULs collected, fixed and processed for histopathologic evaluation. Ultrasonography showed treated SULs had a smaller percent lesion at the maximal injury zone (p = 0.001), and the total percent lesion (p = 0.01), total fiber alignment score (p = 0.0167) and total echogenicity (p = 0.0023) were all smaller in the treated ligaments. The fiber alignment score and echogenicity were not statistically different at the maximal injury zone. The treated SULs had a more concentrated area of metachromasia associated with healing than the untreated ligaments, where metachromasia appeared more diffuse throughout the ligament. ESWT improved the rate of healing as assessed ultrasonographically. PMID- 15121249 TI - Noninvasive bladder volume measurements based on nonlinear wave distortion. AB - The purpose of this study is to suggest a new approach to measure noninvasively the fluid content of a human cavity; in particular, the bladder volume. Determination of bladder volume is important clinically for patients suffering from bladder dysfunction. Several ultrasonic methods were proposed in the past for such a purpose. Most of these methods are based on multiple cross-sectional images and detection of echoes from the bladder wall. We propose here, in a first step, a simple approach that is able to provide an indication of whether or not the bladder volume has exceeded a certain threshold volume. Second, the possibility of accurate volume assessment will be discussed. The approach takes advantage of the difference in harmonic generation of liquids (urine) and tissues. We know that nonlinear effects occur most strongly when ultrasound (US) propagates through liquids with relatively low acoustic attenuation, such as water or urine. However, within soft tissues, the tendency for wave distortion to occur is limited as a result of different acoustic characteristics, most notably the attenuation. Our method is based on measuring the presence of harmonics in an echo from a region-of-interest (ROI) at fixed depth beyond the bladder. The harmonic content in the echo will increase with increase of fluid in the echo path. Phantom measurements were carried out with a single-element transducer and a phased-array transducer. Phantoms containing different volumes were used. Furthermore, measurements on volunteers were performed. The results confirmed that the harmonic content of an echo measured at a deep ROI increases for a full bladder and decays strongly after the volunteer has voided. These preliminary results demonstrate the feasibility of the approach. PMID- 15121250 TI - Wavelet denoising of displacement estimates in elastography. AB - Wavelet shrinkage denoising of the displacement estimates to reduce noise artefacts, especially at high overlaps in elastography, is presented in this paper. Correlated errors in the displacement estimates increase dramatically with an increase in the overlap between the data segments. These increased correlated errors (due to the increased correlation or similarity between consecutive displacement estimates) generate the so-called "worm" artefact in elastography. However, increases in overlap on the order of 90% or higher are essential to improve axial resolution in elastography. The use of wavelet denoising significantly reduces errors in the displacement estimates, thereby reducing the worm artefacts, without compromising on edge (high-frequency or detail) information in the elastogram. Wavelet denoising is a term used to characterize noise rejection by thresholding the wavelet coefficients. Worm artefacts can also be reduced using a low-pass filter; however, low-pass filtering of the displacement estimates does not preserve local information such as abrupt change in slopes, causing the smoothing of edges in the elastograms. Simulation results using the analytic 2-D model of a single inclusion phantom illustrate that wavelet denoising produces elastograms with the closest correspondence to the ideal mechanical strain image. Wavelet denoising applied to experimental data obtained from an in vitro thermal lesion phantom generated using radiofrequency (RF) ablation also illustrates the improvement in the elastogram noise characteristics. PMID- 15121251 TI - Ultrasound attenuation in normal and spontaneously degenerated articular cartilage. AB - High-frequency ultrasound (US) measurements may provide means for the quantification of articular cartilage quality. Bovine patellar cartilage samples (n = 32) at various degenerative stages were studied using US attenuation measurements in the 5- to 9-MHz frequency range. The results were compared with the histologic, biochemical and mechanical parameters obtained for the same samples, to identify which structural or functional factors could be related to the attenuation and its variations. Attenuation, as calculated in the frequency or time domain, correlated significantly with the histologic tissue integrity (i.e., Mankin score, Spearman r = -0.576 or -0.571, p < 0.01), but the slope of attenuation vs. frequency was not related to Mankin score. Ultrasound speed was, however, the most sensitive indicator of Mankin score (r = -0.755, p < 0.01). Cartilage quality index (CQI), a combination of structural and functional parameters, correlated significantly with the attenuation or speed (r = -0.655 or -0.872, p < 0.01). Our results suggest that US attenuation and speed may be suited for the diagnostics of cartilage degeneration. (E-mail: ) PMID- 15121252 TI - Moderately nonlinear ultrasound propagation in blood-mimicking fluid. AB - In medical diagnostic ultrasound (US), higher than-in-water nonlinearity of body fluids and tissue usually does not produce strong nonlinearly distorted waves because of the high absorption. The relative influence of absorption and nonlinearity can be characterized by the Gol'dberg number Gamma. There are two limiting cases in nonlinear acoustics: weak waves (Gamma < 1) or strong waves (Gamma >> 1). However, at diagnostic frequencies in tissue and body fluids, the nonlinear effects and effects of absorption more likely are comparable (Gol'dberg number Gamma approximately 1). The aim of this work was to study the nonlinear propagation of a moderately nonlinear US second harmonic signal in a blood mimicking fluid. Quasilinear solutions to the KZK equation are presented, assuming radiation from a flat and geometrically focused circular Gaussian source. The solutions are expressed in a new simplified closed form and are in very good agreement with those of previous studies measuring and modeling Gaussian beams. The solutions also show good agreement with the measurements of the beams produced by commercially available transducers, even without special Gaussian shading. PMID- 15121253 TI - Circulating tumor cells in patients with solid malignancy treated by high intensity focused ultrasound. AB - The theoretical possibility that exposure of a solid malignancy to high-intensity focused ultrasound (US), or HIFU, could lead to an increased rate of metastasis still remains. Using reverse transcriptase polymerase chain reaction (RT-PCR), the potential risk of hematogenous dissemination was assessed in HIFU-treated patients with solid malignancy. RT-PCR can demonstrate the presence or absence of specific RNA fragments. On the day before HIFU ablation, 5-mL peripheral blood samples were collected, and again 5 to 7 days after HIFU, from 26 enrolled patients (hepatocellular carcinoma, HCC: 10; osteosarcoma: 16). Total RNA was isolated and RT-PCR was performed to analyze the mRNA expression of (alpha fetoprotein (AFP) and bone-specific alkaline phosphatase (BALP) genes. Positive AFP mRNA expression was preoperatively detected in 8 of 10 patients with HCC. In the postoperative specimens, positive expression was also detected in 8 of 10 patients. In 2 patients, circulating tumor cells were found preoperatively, but not postoperatively. Conversely, 2 patients with no circulating tumor cells preoperatively were found to have circulating tumor cells after HIFU. Of 16 osteosarcoma patients, 12 patients had circulating tumor cells and 4 had none. After HIFU treatment, 2 of the 12 patients had converted from presence to absence of circulating cells and the remaining 4 patients remained negative. It is concluded that patients undergoing complete HIFU ablation may demonstrate conversion from presence to absence of circulating tumor-specific marker mRNA, and that HIFU would not enhance the potential risk of metastasis in patients with malignant diseases. PMID- 15121254 TI - Ultrasound-induced cell membrane porosity. AB - Recent studies of ultrasound (US) methods for targeted drug delivery and nonviral gene transfection revealed new, advantageous possibilities. These studies utilized US contrast agents, commonly stabilized microbubbles, to facilitate delivery and suggested that US delivery resulted from cell sonoporation, the formation of temporary pores in the cell membrane induced by US. Using voltage clamp techniques, we obtained real-time measurements of sonoporation of single Xenopus oocyte in the presence of Optison trade mark, an agent consisting of albumin-shelled C(3)F(8) gas bubbles (mean diameter 3.2 microm). Ultrasound increased the transmembrane current as a direct result of decreased membrane resistance due to pore formation. We observed a distinct delay of sonoporation following US activation and characteristic stepwise increases of transmembrane current throughout US duration. We discovered that the resealing of cell membrane following US exposure required Ca(2+) entering the cell through US-induced pores. PMID- 15121255 TI - Physical parameters influencing optimization of ultrasound-mediated DNA transfection. AB - Ultrasound (US) has been shown to transiently disrupt cell membranes and, thereby, facilitate the loading of drugs and genes into viable cells. To address optimization of gene therapy applications, the aim of this work was to systematically determine the influence of physical parameters on transfection and viability of DU145 prostate cancer cells by two different DNA plasmids (pEGFP-N1 and pGL3). By sonicating cells in vitro in the presence of naked DNA, we found that transfection efficiency was increased by: 1. optimizing acoustic energy at 10 to 30 J/cm(2) (for our apparatus, at pressures above the cavitation threshold); 2. using 500-kHz US in the presence of Optison to nucleate cavition, rather than 24-kHz US without Optison; 3. increasing cell concentration from 10(6) to 10(7) cells/mL; and 4. changing temperature during sonication from 21 to 37 degrees C. The best conditions in this study increased transfection by almost 100-fold in the absence of significant DNA damage. Additional measurements indicated that less than one fourth of cells with DNA plasmid uptake into the cytosol showed DNA expression, which suggests that further optimizing transfection by US may require facilitating intracellular DNA trafficking. PMID- 15121256 TI - Effects of ultrasound-targeted microbubble destruction on cardiac gene expression. AB - Ultrasound (US) contrast agents are increasingly used in diagnostic echocardiography. Recent studies have suggested unanticipated effects of microbubble destruction. This study was designed to evaluate gene regulation caused by US-mediated destruction of microbubbles in the heart. During IV infusion of Optison trade mark, triggered US was applied to rat hearts to destroy microbubbles. A control group received only saline and US. RNA was isolated from hearts 24 and 72 h after treatment. Analysis with a deeply representative murine cardiac-specific microarray was used to identify regulated genes. Real-time polymerase chain reaction (PCR) was then applied to verify regulated genes. Microarray analysis revealed only 5 regulated genes in the 24-h group and 4 in the 72-h group. Of these genes, only carbonic anhydrase was significantly upregulated in the 24-h Optison trade mark group (4.3 fold; p = 0.0005) when examined in individual animals by real-time PCR. By this very sensitive technique, the bioeffects of microbubble destruction are negligible. PMID- 15121257 TI - Investigation of genotoxic effect of ultrasound in cases receiving therapeutic ultrasound by using micronucleus method. AB - In 1991, reported that therapeutic ultrasound (US) did not induce sister chromatid exchanges (SCEs) in patients whereas, in 1984, reported that each of 10 patients exposed to therapeutic US had a statistically significant increase in SCEs. The present study was planned to investigate if there was chromosomal damage resulting from therapeutic US by using a micronucleus (MN) method, and to counter the lack of reports in this area over the past 10 years. A total of 20 female volunteers were included in the study; 10 of them with low back pain (mechanical low back pain and facet syndrome) were treated with US and 10 healthy cases constituted the control group. Patients with low back pain received 10 sessions of US therapy at an intensity of 2 W/cm(2) and a frequency of 1 MHz for 10 min and patients in the control group received sham US therapy for 10 min. Peripheral blood taken before and after the fifth and tenth applications of US therapy was cultured for MN frequencies both for the treatment and the control groups. The scores of MN assessed before the therapy were compared with those at the end of the fifth session and the end of the tenth session in the treatment and the control groups. Pretreatment, end of the fifth session and end of the tenth session MN frequencies were compared between the treatment and the control groups. There was no statistically significant difference in MN frequencies between pretreatment and fifth session or pretreatment and tenth session in both groups. Nor was there any significant difference in the MN frequencies of the treatment and control groups between pretreatment, fifth session and tenth session evaluations. In conclusion, we observed that therapeutic US did not induce increases in MN frequency, which are a sign of cytogenetic damage. PMID- 15121258 TI - Effects of diagnostic cardiac ultrasound on oxygen free radical production and microvascular perfusion during ischemia reperfusion. AB - Diagnostic ultrasound (US) is reported to increase intracellular oxidative stress in vitro. Increased oxidative stress mediated ischemia-reperfusion injury in the microcirculation. To examine the effects of US in hamster cheek pouch microcirculation during baseline and ischemia and reperfusion (I/R), I/R injury was provoked in the cheek pouch under "sham" (transducer off, group 1) and active US irradiation (group 2) at baseline (15 min) and at the beginning (15 min) of the reperfusion after ischemia (30 min). US transmission was delivered in the harmonic mode (2.5 MHz) with 1.3 mechanical index (MI) and 2.0 peak negative pressure. Microvascular damage was evaluated by measuring arterial diameter, red blood cell velocity, wall shear stress, permeability, perfused capillary length and adherent leukocytes in venules. Lipid peroxides were determined in the systemic blood. US increased permeability (baseline: 0.04 +/- 0.02; after US 0.30 +/- 0.04, p < 0.01) and slightly decreased capillary perfusion by 7% during baseline (p < 0.01). Arteriolar diameter (35 +/- 7 microm vs. 20 +/- 5 microm, p < 0.05), RBC velocity (2.8 +/- 0.4 mm s(-1) vs. 0.75 +/- 0.05 mm s(-1), p < 0.05) and shear stress ( 0.76 +/- 0.09 Pa vs. 0.36 +/- 0.05 Pa, p < 0.05) decreased significantly after reperfusion. These parameters increased by 40, 64 and 33%, respectively after US. Leukocyte adhesion decreased by 31 % (p < 0.05) after US and lipid peroxides decreased by 26% and 51% during baseline and 15 min of reperfusion after US, respectively. In conclusion, diagnostic US increased microvascular permeability during baseline and reperfusion. Moreover, US enhanced wall shear stress and reduced oxidative stress during postischemic reperfusion; thus, increasing capillary perfusion. PMID- 15121259 TI - Acoustic streaming: an in vitro study. AB - The aims of this study were, first, to determine if cyst size and cyst-to transducer distance have an impact upon acoustic streaming and, second, to investigate the effect of cyst content viscosity on acoustic streaming using an artificial ovarian cyst model. Artificial ovarian cysts were constructed and suspended in a tissue-mimicking bath. Although there was no subjective difference in acoustic streaming velocity between cyst sizes during B-mode insonation, with colour Doppler and pulsed Doppler examination, higher acoustic streaming scores were recorded for larger cysts. When the cyst-to-transducer distance was increased, the acoustic streaming velocity was noted to decrease in all scanning modalities. The second stage of the study demonstrated decreasing acoustic streaming velocity as the viscosity of the cyst content was increased. The finding of a clear association between cyst content viscosity and acoustic streaming velocity raises the exciting possibility that we may be able to make estimations of the viscosity of ovarian cyst contents, in the clinical setting, by sonographic means. PMID- 15121260 TI - Squamous cell cancer of the buccal mucosa in young adults. AB - BACKGROUND: Squamous cell carcinoma of the buccal mucosa is predominantly a cancer of elderly people and is seen rarely in young adults. Although many aetiological factors have been postulated, tobacco alone has been implicated as the main factor in the aetiology of such cancers at any age. PATIENTS AND METHODS: A retrospective analysis of 46 patients with cancer of the buccal mucosa who were under the age of 35 years at the time of presentation was made to evaluate their survival and predictors of survival. The details were extracted from the computerised records and case records of the Regional Cancer Centre, Trivandrum, India. These were analysed with particular emphasis on risk factors, clinical extent, and treatment. Survival was analysed by the Kaplan-Meier method, and differences in survival were compared using the log-rank test. RESULTS: The male:female ratio was 6.7:1. Almost all the patients (n = 42, 91%) used either tobacco or alcohol, with tobacco chewing being the most common. The 3 and 5 years disease-free survival were 72 and 61%, respectively. Non-smokers and patients with an advanced T stage at presentation had significantly worse prognoses. PMID- 15121261 TI - Evaluation of speech intelligibility after a secondary dehiscence operation using an artificial graft in patients with speech disorders after partial glossectomy. AB - The purpose of this study was to evaluate speech intelligibility after a dehiscence operation using artificial grafts for patients with speech disorders after partial glossectomy that were caused by scars resulting from the primary operation. The subjects were six men and three women, who had had a partial glossectomy for tongue cancer followed by direct closure without reconstruction. They were operated on a second time operation to mobilise the residual tongue by dividing the cicatrix. An artificial graft was applied to the wound to maintain the dehiscence. Speech intelligibility was evaluated by a standardised Japanese speech intelligibility test before, and 6 and 12 months after the second operation. The intelligibility scores significantly improved during the first 6 months after the second operation, and continued to improve slightly during the following 6 months. This study suggests that the dehiscence operation using an artificial graft could improve speech in patients after partial glossectomy. PMID- 15121263 TI - The CRABEL score--setting standards in maxillofacial medical note-keeping. AB - The CRABEL score (developed by Crawford, Beresford and Lafferty) was introduced for auditing medical note-keeping at Morriston Hospital in June 2001. Guidelines detailing the scoring system were issued to all clinicians in the maxillofacial unit. An auditor selected two sets of medical notes from each consultant's firm, giving an initial allocation of 100points/firm (50 points for each set of notes). The notes of the most recent in-patient admission were analysed using the CRABEL marking sheet to give a score out of 100 for each firm. The audit was repeated at 3-month-intervals. CRABEL scores within the maxillofacial unit improved from 70 to 97. The CRABEL score is simple, reliable and repeatable. It is a successful and objective measure for audit and for improvement in the quality of note keeping. We propose that it be adopted in maxillofacial units throughout the United Kingdom. PMID- 15121262 TI - Polymorphous low-grade adenocarcinoma--a rare and aggressive entity in adolescence. AB - Polymorphous low-grade adenocarcinoma (PLGA) is an uncommon tumour that usually affects the minor salivary glands, particularly in the palate. It is rare in young patients, and here we report a case in a teenage girl. She presented at the age of 16, although the lesion had been noticed 2 years previously. The tumour showed histopathological features of PLGA, but recurred locally, behaved aggressively, and ultimately metastasised to cervical lymph nodes. This was accompanied by an altered histological picture, with a papillary cystic pattern and necrosis becoming progressively more prominent. PLGA is not always a low grade lesion and some tumours, notably those with a papillary cystic growth pattern, may require more aggressive treatment. PMID- 15121264 TI - Removal of partially erupted third molars using an Erbium (Er):YAG laser: a randomised controlled clinical trial. AB - We compared Erbium (Er):YAG laser with a surgical bur for removal of partially erupted lower third molars. Patients were randomised to be treated by either laser or bur. A total of 42 patients (laser = 22; bur = 20) were treated. A greater reduction in the range of mouth opening was found after laser than after bur treatment. Postoperative pain was more common after bur treatment. The duration of operation was considerably longer with laser than with bur. No persistent complications were encountered. PMID- 15121266 TI - Effects of low-dose midazolam with propofol in patient-controlled sedation (PCS) for apicectomy. AB - We studied the effects of low-dose midazolam with propofol for patient control sedation (PCS) in 30 healthy (ASA grade I) patients who were randomly allocated into two equal groups (n = 15 in each). They were given a propofol infusion of 2mg/kg/h after a bolus dose of 0.7 mg/kg. The second group was given the 2mg/kg/h propofol infusion after a dose of midazolam 0.03 mg/kg and a bolus dose of propofol 0.7 mg/kg. The standard dose for PCS was propofol 0.2mg/kg in both groups. Clinical data were taken and haemodynamic variables, and oxygen saturation were recorded before and on the 5th, 10th, 20th, and 30th minutes during the operations. The level of sedation, amnesia and conditions of each patient were evaluated during the study. Patients' satisfaction was recorded using a modified visual analogue scale (VAS). All results were evaluated statistically. We conclude that low-dose midazolam with propofol during PCS neither reduced oxygen saturation nor prolonged the time of discharge. Low-dose midazolam with propofol also improved the acceptability and comfort for patients and made the operation easier, which makes it preferable to propofol alone. PMID- 15121265 TI - Clinical postoperative findings after removal of impacted mandibular third molars: prediction of postoperative facial swelling and pain based on preoperative variables. AB - PURPOSE: This paper is intended as an investigation of the relationship between preoperative findings and short-term outcome in third molar surgery. MATERIAL AND METHODS: We assessed 153 consecutive surgical extractions of mandibular third molars performed in 140 patients between April 1998 and March 2001. RESULTS: Fifty-four (35%) of the 153 extractions were performed in male subjects and 99 (65%) in female subjects. The median age was 27 years. The amount of facial swelling varied depending on age and sex. Severe pain was associated with depth and preoperative index of difficulty. Average pain was associated with preoperative index of difficulty. CONCLUSION: In conclusion, we consider that the short-term outcomes of third molar operations (swelling and pain) differ depending on patients' characteristics (age and sex) and preoperative index of difficulty. Further mega-trial studies of the association between preoperative findings and short-term outcome will help to elucidate the true nature and magnitude of the association. PMID- 15121267 TI - The skeletal stability of one-piece Le Fort 1 osteotomy to advance the maxilla; Part 1. Stability resulting from non-bone grafted rigid fixation. AB - During the past decade, we have increasingly preferred to do a one-piece Le Fort 1 osteotomy to advance the maxilla, sometimes in isolation to treat patients with maxillary retrusive skeletal Class III patients or combined with mandibular advancement to treat bimaxillary retrusive skeletal Class II. Clinical impressions of rigid fixation techniques have indicated that there is improved stability when compared with wire fixation. There are few studies in the literature that have addressed relapse following one-piece Le Fort 1 osteotomy to advance the maxilla. Such surgery involves one single spatial movement and thereby eliminates other possible surgical variables, which may impact on the degree of stability achievable postoperatively. We studied 45 patients who had undergone a uniform one-piece maxillary advancement with elimination of controllable variables, apart from 15 patients who had simultaneous mandibular advancement. Rigid fixation was adopted throughout the study. The mean surgical change documented was 7.42 mm. The mean stability calculated at 12 months revealed a relapse of 0.72 mm (10%). This was not significant (P = 0.3). We conclude that the Le Fort 1 advancement osteotomy is a stable and surgically predictable procedure that gives only slight relapse at 12 months. PMID- 15121268 TI - The skeletal stability of one-piece Le Fort 1 osteotomy to advance the maxilla; Part 2. The influence of uncontrollable clinical variables. AB - The stability of orthognathic surgery has been the subject of numerous publications over the last 20 years. It is now apparent that studies must pay attention to the homogeneity of the patients investigated and in particular, surgical techniques. In Part 2 of our study of 45 patients who had a uniform one piece maxillary advancement with rigid fixation to advance the maxilla, we found that uncontrollable variables, including patients age, sex, the degree of advancement, and simultaneous mandibular advancement, had no effect on post operative skeletal stability. PMID- 15121269 TI - Blood transfusion requirements in bimaxillary osteotomies. AB - The over-ordering of cross-matched blood to cover operations can result in blood shortages and is costly; it can never be free of risk. Current published guidelines recommend cross-matching 2 units of blood for bimaxillary orthognathic procedures with an additional 2 units if combined with a genioplasty. We reviewed the records of 115 consecutive cases of simultaneous bimaxillary osteotomies at Morriston Hospital over a 5-year period (January 1996 to December 2000). Ordering and use of blood were investigated and the cost analysed. Blood loss was minimised using a strategy of controlled moderate hypotension and meticulous haemostasis. Nine patients were given transfusions of blood but five of these were deemed inappropriate. No predisposing factors for transfusion were identified. We recommend that the tariff for ordering blood for bimaxillary osteotomies should be revised to a "group and save" with antibody screen, providing that a 30-min indirect antibody cross-match is available. PMID- 15121270 TI - A minimally invasive novel design for a vascular-pedicled bone segment for experimental studies of reconstruction of mandibular defects. AB - We describe a new minimally invasive vascularised bone segment (VBS) for experimental reconstruction of the mandible. The study was validated and the feasibility was tested on fresh sheep cadavers. The integrity of the arterial blood supply was confirmed using inferior dental artery static arteriography. A pilot investigation was conducted on two adult sheep. Two full thickness bone cuts 35 mm apart were created in the parasymphyseal region. The inferior dental neurovascular bundle was preserved at the posterior bone cut but not anteriorly. A VBS was created, pedicled only on the inferior dental artery. The continuity of the mandible was maintained using fixation plates and the animals were followed up for 12 weeks. There was complete bony union and restoration of approximately 80% of the bony strength. The model is minimally invasive, reduces the operating time, and does not require special equipment and skills in microvascular anastomosis. PMID- 15121271 TI - Reconstruction of the temporomandibular joint by the transfer of the free vascularised second metatarsal. AB - We present our experience of a series of patients who presented for salvage reconstruction of the temporomandibular joint (TMJ) for relief of pain. Reconstruction was achieved by transfer of the free vascularised second metatarsal. This technique has been used for a total of seven TMJ reconstructions in five patients. We describe the surgical anatomy, technique and results during the last 18 years. One joint failed but the other six surviving joints continue to provide adequate pain-free function. We advocate this technique for autogenous salvage reconstruction in joints that have been previously operated on unsuccessfully. PMID- 15121272 TI - Interleukin-6 family of cytokines as biochemical markers of osseous changes in the temporomandibular joint disorders. AB - The aim of this study was to find out whether there was a correlationship between the concentrations of interleukin-6 (IL-6), IL-11, and IL-17 in synovial fluid and osseous changes in the condyle. The synovial fluid was obtained from 61 patients with temporomandibular joint disorders (TMD) and seven healthy volunteers (controls). The concentrations of IL-6, IL-11, and IL-17 were measured by enzyme-linked immunosorbent assay. IL-6 was detected in 43 of 59 (73%), IL-11 in 23 of 52 (44%) and IL-17 in 14 of 51 (27%) samples of synovial fluid. The concentrations of IL-6 and IL-11 in the joints with osseous changes in the condyle were significantly higher than in the joints without osseous changes (P < 0.05) and also higher than in the joints of the control group (P < 0.05). In addition, there was a correlation of concentrations between IL-6 and IL-11 (P < 0.05). These results suggest that IL-6 and IL-11 may participate in the pathogenesis of TMD and induce osseous changes in the condyle. PMID- 15121273 TI - Venous lakes of the vermillion lip treated by infrared coagulation. AB - We reviewed the treatment of 20 venous lakes by infrared coagulation in 18 patients. Seventeen cleared after one treatment; in three patients a further treatment was required, and one patient needed a total of three sessions to clear the venous lake. At 1-6 months follow-up there was complete clearance with no discernible mark in all but four patients who had minimal scarring, including the patient who had three treatments. PMID- 15121274 TI - Titanium plate reconstruction of the osseous defect after harvest of a composite free flap using the deep circumflex iliac artery. AB - Hernia formation following harvest of bicortical iliac crest bone occurs infrequently as a late complication and may lead to chronic pain at the donor site and rarely to obstruction and strangulation of bowel. We describe the use of a custom-made titanium plate used to reconstruct the iliac donor site following harvest of a DCIA composite free flap. A pre-operative 3D CT and stereolithography model of the ilium are used to fabricate a titanium plate of the desired shape and size. This plate is used to reconstruct the donor site defect at the time of primary surgery. This technique may reduce late complications following DCIA composite free flap harvest. PMID- 15121275 TI - Polyvinylsiloxane dental impression material used to support the pinna after severe injury. AB - This case illustrates the simple, cheap and immediate provision of a splint for the pinna after severe injury using dental polyvinylsiloxane impression material. PMID- 15121276 TI - Synovial chondromatosis of the temporomandibular joint: clinical and immunohistopathological considerations. AB - A histopathological study of 30 cases of synovial osteochondromatosis found that the process followed a temporal sequence characterised by three phases: (I) active intrasynovial disease only; (II) transitional lesions with both active intrasynovial proliferation and free loose bodies; and (III) many free osteochondral bodies with no demonstrable intrasynovial disease [J. Bone Joint Surg. 59 (1977) 792]. We present five cases of synovial chondromatosis of the temporpmandibular joint (TMJ) which we studied by immunohistochemical methods of for transforming growth factor beta (TGFbeta) and tenascin. PMID- 15121277 TI - Synovial sarcoma of the hard palate. AB - Synovial sarcoma is a rare soft tissue tumour of uncertain histogenesis and intraorally they occur mainly in young adults. We present a case of synovial sarcoma of the hard palate in a 68-year-old woman. To our knowledge, there is no previously reported instance of this tumour at this site. PMID- 15121278 TI - Axillary metastases from recurrent oral carcinoma. AB - The rationale for surgical treatment of head and neck cancer is based on a predictable pattern of metastasis. There is aberrant or unpredictable spread rarely and typically only in recurrent disease. There are few published reports to our knowledge of axillary metastases from squamous cell cancer (SCC) of the head and neck. We present a patient who developed axillary node disease on the other side after recurrence of a squamous cell carcinoma of the floor of the mouth. She died 11 months after excision of the recurrence. PMID- 15121279 TI - Cancrum oris. AB - A malnourished 9-year-old boy presented with an infection in the buccal space that developed into cancrum oris during the course of treatment. PMID- 15121280 TI - Coincidental finding of a bipartite atlas during assessment of facial trauma. AB - We report a patient with a bipartite atlas that was noted on preoperative skull radiographs for fractures of facial bones. PMID- 15121281 TI - Reduction of a chronic bilateral temporomandibular joint dislocation with intermaxillary fixation and botulinum toxin A. AB - A 71-year-old man was referred to the Department of Oral and Maxillofacial Surgery at Westmead Hospital for investigation of limited jaw movement and facial pain after a cerebrovascular event eight weeks previously. He was found to have bilateral dislocations of the temporomandibular joints (TMJ) and was successfully treated with a combination of intermaxillary fixation (IMF) screws and botulinum toxin A. PMID- 15121282 TI - Re: Malden NJ, The use of the laryngeal mask in minor oral surgery (Br J Oral Maxillofac Surg 2003;41:343-5). PMID- 15121283 TI - Re: Silva EC, Souza PEA, Barreto DC, Dias RP, Gomez RS, An extreme case of cherubism (Br J Oral Maxillofac Surg 2002;40:45-8). PMID- 15121286 TI - Digital photography in the management of maxillofacial trauma. PMID- 15121287 TI - Re: Debate about the microbiology of epidural spinal abscesses of dental origin. PMID- 15121288 TI - Re: Training in maxillofacial surgery. PMID- 15121289 TI - Fast tracking of referrals for orthodontic oral surgery. PMID- 15121290 TI - Chimpanzees and journalists. PMID- 15121291 TI - Oral polio vaccine: fact versus fiction. AB - The author of this article has now become the principal target of the accusation that HIV-1 was introduced into man through the wildcat production of an oral polio vaccine in chimpanzee cell culture more than 40 years ago in the Belgian Congo. The putative evidence presented in support of this accusation rests essentially on testimonials by local witnesses. Such an accusation is strongly rejected on basis of a factual review of the author's career and his publications, as well as his total incapability to have managed such a feat at the supposed time. PMID- 15121292 TI - Material deprivation, hospitalisation and adverse events temporally associated with immunisation. AB - We examined hospital admissions for illness temporally associated with childhood immunisations to determine the influence of material deprivation. Children aged <5 years hospitalised in the West Midlands were identified using ICD10 codes (Y580-Y599) from April 1995 through March 2000. Material deprivation was measured using the Townsend score. Children from deprived areas were at increased risk (OR = 4.49, 95% CI 3.14-7.78) compared to children from affluent areas and, were more likely to be admitted following immunisation with pertussis vaccine (OR = 5.91, 95% CI 2.69-12.29). Further research is required to determine the contribution of healthcare providers and parents to this health differential. PMID- 15121293 TI - Itching nodules and hypersensitivity to aluminium after the use of adsorbed vaccines from SSI. PMID- 15121294 TI - Construction of recombinant pseudorabies virus expressing NS1 protein of Japanese encephalitis (SA14-14-2) virus and its safety and immunogenicity. AB - The bivalent genetic engineering vaccine of Japanese encephalitis (JE) and Aujeszkj disease (AD) was developed to provide a novel approach to prevent and control these two diseases. NS1 gene of Japanese encephalitis virus (JEV) SA14-14 2 strain was produced by reverse transcriptase-mediated PCR (RT-PCR) and was cloned into vector pUSK to form recombinant plasmid (designed as pUSK-NS1). A co transfection experiment was performed in porcine kidney (PK-15) cells with pUSK NS1 and the genome of the vector virus (PRV TK(-)/gG(-)/LacZ(+) mutant). By plaque purification, PCR detection and southern hybridization, recombinant pseudorabies virus (PRV) expressing NS1 protein of JEV was acquired and named TK( )/gG(-)/NS1(+). Western blot analysis and ELISA demonstrated the NS1 protein expression. To evaluate the recombinant virus's potential application, we characterized the safety and immune responses in Balb/c mice and swine. The safety test indicated that, when receiving the recombinant virus at a concentration of 10(6.0)pfu, no virulence of the recombinant virus to the mice, piglets and pregnant sows was observed. The vaccinated animals could acquire protective immunity against lethal challenge of the virulent PRV Ea strain and develop a good humoral and cellular immune response against JEV. The above results revealed that the recombinant virus could be a suitable candidate vaccine strain for developing a novel genetic vaccine to combat pseudorabies and Japanese encephalitis in the pig industry. PMID- 15121295 TI - The evidence for the safety of thiomersal in newborn and infant vaccines. AB - While a number of studies remain to be completed, evidence is mounting that there is no demonstrable risk for infants immunized with vaccines containing thiomersal. Epidemiological studies in the US have shown no developmental or other central nervous system abnormalities resulting from exposure to vaccines containing thiomersal. During the initial evaluation of thiomersal in vaccines during 1999, the toxicological profile of ethyl mercury was unknown and presumed to be the same as that of methyl mercury. Enough evidence has accumulated since then to indicate the profiles of the two compounds are different in crucial aspects. To date, one study has measured blood levels of total mercury in vaccinated infants and reports only a brief low-level exposure with rapid excretion of mercury. It is not yet known for sure how much (if any) vaccine derived ethyl mercury in the blood crosses the blood-brain barrier. For the most part, the use of thiomersal as a vaccine preservative has been convincingly shown to be safe. The scientific evidence is not yet sufficiently strong to provide the same level of assurance for thiomersal-containing vaccines for use in pregnant women or the premature or low birth weight infant. There is an increased sensitivity of the fetal brain to mercury whether it is ethyl or methyl mercury. While there is no evidence to support the contention, it is at least theoretically possible that very low birth weight premature infants may be at increased risk from thiomersal-containing vaccines. Until such time as the scientific evidence is to hand, thiomersal-free presentations of hepatitis B are to be preferred for the birth dose. Given the same levels of exposure, adults are at much lower levels of risk because of increased body mass. It is not possible to prove that thiomersal is completely safe-epidemiology can only quantify a risk, not prove its absence. PMID- 15121296 TI - Pharmaco-economic evaluation of targeted hepatitis A vaccination for children of ethnic minorities in Amsterdam (The Netherlands). AB - OBJECTIVE: Estimate cost-effectiveness of vaccination against hepatitis A virus (HAV) for children of ethnic minorities in Amsterdam. BACKGROUND: Pharmaco economic analysis is relevant for motivating reimbursement of vaccination costs in the framework of a programmatic approach to vaccination of ethnic minorities. DESIGN: Pharmaco-economic modeling. METHOD: In cost-effectiveness analysis, costs, benefits and health gains were estimated for a large-scale HAV-vaccination for children of Turkish and Maroccan origin. Analysis was performed from the societal perspective, as recommended in the Dutch guidelines for pharmaco economic research. This implies that indirect costs of production losses are included in the analysis. Cost-effectiveness was expressed in net costs per adult HAV-infection averted in incremental and aggregate analysis. Incremental analysis compares targeted vaccination with the current limited-scale HAV-vaccination that exists, whereas aggregate analysis compares targeted vaccination with the sheer absence of vaccination. RESULTS: Net aggregate costs of targeted HAV-vaccination for Turkish and Maroccan children in Amsterdam amounts to 61.000. Cost effectiveness was estimated, in aggregate and incremental analysis, at 13.500 and 11.100 respectively per adult HAV-infection averted. Uni- and multivariate sensitivity analyses show that major impact on cost-effectiveness may be expected from reductions in the vaccine price through economies of scale. Probabilistic sensitivity analysis indicates possible large fluctuations in cost-effectiveness from 1 year to another, related to varying incidence of disease. CONCLUSION: HAV vaccination for children from ethnic minorities in Amsterdam is not cost saving, but may have a favourable cost-effectiveness. Such a vaccination program fits into the recent Dutch policy of specific vaccinations directed at groups of ethnic minorities, such as for hepatitis B. PMID- 15121297 TI - Acute Trypanosoma cruzi infection: IL-12, IL-18, TNF, sTNFR and NO in T. rangeli vaccinated mice. AB - We have developed an experimental model of vaccination against the infection with the protozoa Trypanosoma cruzi, the agent of Chagas disease in Latin America. Vaccination was performed with Trypanosoma rangeli, a non-pathogenic protozoa sharing many antigens with T. cruzi. It strongly protected BALB/c mice, sharply reducing parasitaemia and mortality rate of the acute T. cruzi infection. The aim of the present work was to complete our previous study on the production of IFN gamma and IL-10 in this vaccination model by investigating the production of IL 12p35 and p40, IL-18, TNF, TNF soluble receptors (sTNFR), and nitric oxide (NO), factors known to play a key role in the outcome of T. cruzi infection. We show that the protection obtained against the acute T. cruzi infection was surprisingly associated with reduced circulating levels of IL-18 and NO, whereas the release of IL-12p40 was enhanced in comparison to non-vaccinated infected animals. IL-12p35 remained undetectable in infected animals, vaccinated or not. The balance between sTNFR and TNF suggested a decrease of TNF bioactivity in vaccinated mice. These results show that the protection induced by the vaccination with T. rangeli against a challenging infection with T. cruzi is not associated with the strong type 1 immune response usually involved in the control of intracellular pathogens, particularly questioning the protective role of NO during the acute phase of T. cruzi infection. PMID- 15121298 TI - Intracellular fate and immunogenicity of B. subtilis spores. AB - To support our work on the development of bacterial spores as oral vaccines we examined the immunogenicity and intracellular fate of Bacillus subtilis endospores in a murine model. Mice dosed orally with spores developed systemic IgG and mucosal sIgA responses. Analysis of IgG subclasses revealed a predominance of the IgG2a subclass during the early stages of immunisation. Analysis of cytokine mRNA in GALT and lymphoid organs showed early induction of IFN-gamma, a Th1 cytokine, as well as the pro-inflammatory cytokine TNF-alpha. Significant levels of IgG antibodies were produced against vegetative bacilli following dosing with spores. This showed that spores could germinate in the GI tract. In vitro studies detailing the intracellular fate and persistence of spores in a macrophage-like cell line (RAW264.7) demonstrated that spores could germinate efficiently in macrophages, initiate gene expression as well as inducing pro-inflammatory cytokines. PMID- 15121299 TI - Evaluation of diphtheria convalescent patients to serve as donors for the production of anti-diphtheria immunoglobulin preparations. AB - AIMS: The study was conducted to evaluate the possibility of selecting convalescent diphtheria patients to serve in emergency situations as donors for the production of anti-diphtheria immunoglobulin. To select suitable donors, the criterion of an antitoxin titer >/=3.0 IU/ml was used. In addition, the effects of treatment and the effect of immunization with diphtheria toxoid on the level of anti-diphtheria toxin antibodies were evaluated. SCOPE: Three groups of diphtheria patients were included in the study. The first group (n = 23) consisted of patients who had a basic antibiotic treatment, with or without serotherapy using horse antitoxin and/or human immunoglobulin. The second group (n = 12) comprised patients examined immediately after the onset of disease. The immunological history of this group was not known. The third group (n = 20) included patients with a known immunization history, treated only with antibiotics but having received a booster immunization with diphtheria toxoid. Antitoxin titers were measured using the toxin binding inhibition (ToBI) assay. CONCLUSIONS: In the first group, 47.8% (11/23) of the patients had a diphtheria antibody titer >/=3.0 IU/ml. For most of them, however, the antibody titers could have resulted from treatment with exogenous antibodies from horse antitoxin or human immunoglobulin (18/23). Only two of the 11 high-titer subjects had received antibiotics only. Among the second group, only two (16.76%) of the patients had an antibody titer of >/=3.0 IU/ml. In the third group 50% (10/20) of the patients showed an antibody titer of >/=3.0 IU/ml prior to vaccination, and therefore could be directly considered as donors. Three weeks after booster vaccination, 70% (14/20) had an antibody titer of >/=3.0 IU/ml and 1 year after booster vaccination, 28.6% (2/7) of the subjects still had titers of >/=3.0 IU/ml. In 40% of these patients, a decrease was observed 3-4 weeks after the booster dose. It was concluded that convalescent diphtheria patients could be considered as donors in an emergency situation, since approximately half of them showed antitoxin titers of >/=3.0 IU/ml. PMID- 15121300 TI - Efficacies of whole cell and acellular pertussis vaccines against Bordetella parapertussis in a mouse model. AB - Pertussis vaccine development has mainly focused on Bordetella pertussis, and consequently these vaccines contain B. pertussis antigens only. However, the related species Bordetella parapertussis can also cause pertussis, although symptoms associated with the disease are generally considered to be milder. Recent field studies have shown that in some outbreaks B. parapertussis can prevail. Using a mouse model we compared the efficacy against B. parapertussis of two commercially available acellular vaccines and two whole cell vaccines, used in The Netherlands and Finland, respectively. The efficacies of the two whole cell vaccines against B. parapertussis were similar, but much lower compared to the efficacy against B. pertussis. Although, the acellular vaccines conferred some protection against B. parapertussis early in infection, the values were not significant. Later in infection, a highly significant enhancement of colonisation by B. parapertussis was observed in mice vaccinated with acellular vaccines. The whole cell vaccines protected significantly better than the acellular vaccines against B. parapertussis. The possible consequences of a switch from whole cell to acellular vaccines was discussed in the light of our findings. PMID- 15121301 TI - The prevalence of anti-rubella antibodies in women of childbearing age in Poland. AB - Congenital rubella is a clinically serious problem, which is due to maternal infection with rubella virus during the first trimester of pregnancy. It may result in spontaneous abortion or in fetal infection leading to fetal birth defect. Maternal anti-rubella antibodies have a protective effect and they may prevent congenital rubella. Vaccination of 13-year-old girls in Poland was introduced in 1989. Vaccination coverage is above 95%. The presence of anti rubella antibodies in maternal blood >15 IU/ml is indicative for immunity to rubella. Thus, the aim of the study was the evaluation of anti-rubella antibody levels in women 15-30 years old. Samples of that sera were collected from six provinces in Poland. We have observed the decrease of anti-rubella antibody titer together with increasing age of tested women. Among 1289 investigated women 135 [10.5%] did not have protective levels of anti-rubella antibodies. We have noticed large decline in protective anti-rubella antibodies in young women (19-22 years of age), who are just entering their prime childbearing years. PMID- 15121302 TI - Interaction of dendritic cells with antigen-containing liposomes: effect of bilayer composition. AB - Vaccine efficacy might be improved by exploiting the potent antigen presenting properties of dendrite cells (DCs), since their ability to stimulate specific major histocompatibility complex-restricted immune responses has been well documented during the recent years. In that light, we investigated how the interaction of antigen-containing liposomes with DCs was affected by the bilayer composition. Monocyte-derived human DCs and murine bone marrow-derived DCs were analysed and compared upon in vitro incubation with liposomes by flow cytometry and confocal microscopy. Anionic liposomes with a bilayer composition of phosphatidylcholine, cholesterol and phosphatidylglycerol or phosphatidylserine interacted with a limited fraction of the total DC population in case of both DC types. Inclusion of mannosylated phosphatidylethanolamine (Man-PE) for targeting to the mannose receptor (MR) increased the interaction of negatively charged liposomes with both human and murine DCs. This increase could be blocked in human DCs by addition of the polysaccharide mannan indicating that uptake might be mediated by the mannose receptor. Cationic liposomes containing trimethyl ammonium propane interacted with a very high percentage of both DC types and could be detected in high amounts intracellularly. In conclusion, liposome bilayer composition has an important effect on interaction with DCs and might be critical for the vaccination outcome. PMID- 15121303 TI - Associations between human leukocyte antigen (HLA) alleles and very high levels of measles antibody following vaccination. AB - Associations between human leukocyte antigen (HLA) genes and very high levels of antibodies (or hyperseroresponsiveness) to measles antigens in a genetically heterogeneous human population are poorly understood. We studied the association between antibody levels after measles vaccination and HLA class I and II alleles among 170 US schoolchildren who received one dose of measles-mumps-rubella II vaccine. Vaccine recipients were divided into two groups: 93 recipients who were seropositive and 77 recipients who were hyperseropositive (the upper 10th percentile of antibody levels of all subjects). Out of all the alleles analyzed, HLA-B(*)7 (odds ratio (OR) 1.9; P = 0.05), DQA1(*)0104 (OR 4.6; P = 0.02) and DPA1(*)0202 (OR 4.8; P = 0.04) alleles were positively associated with hyperseropositivity, whereas HLA-B(*)44 (OR 0.4; P = 0.02), DRB1(*)01 (OR 0.6; P = 0.09), DRB1(*)08 (OR 0.3; P = 0.04), DQB1(*)0301 (OR 0.5; P = 0.04), and DPB1(*)0401 (OR 0.6; P = 0.03) alleles were negatively associated with hyperseropositivity. The alleles B(*)44, DRB1(*)01, DRB1(*)08 and DQA1(*)0104 remained statistically significant after accounting for the effects of other alleles. The results suggest that HLA alleles have important associations with measles antibody hyperseropositivity. These data increase our understanding of measles vaccine-induced immune response and will be useful for future mechanistic work on measles virus antigen processing and presentation in seronegative and hyperseropositive individuals. PMID- 15121304 TI - Development of a hemi-nested RT-PCR method for the specific determination of European Bat Lyssavirus 1. Comparison with other rabies diagnostic methods. AB - A simplified hemi-nested reverse transcriptase polymerase chain reaction (hnRT PCR) has been developed to determine specifically the European Bat Lyssavirus 1 (EBLV-1) nucleoprotein gene. The specificity of this method was determined by using the seven genotypes of lyssavirus by RT-PCR, Southern blot and sequence analysis. Compared to the rabies diagnostic methods, the hnRT-PCR showed a higher sensitivity for the detection of small amounts of EBLV-1 virus. In view of these results, we suggest this new hnRT-PCR should be performed for the epidemiological survey of bat colonies, also providing rapid detection and genotyping of EBLV-1 until now encountered in all naturally infected bats in France. PMID- 15121305 TI - Immunization with the hybrid protein vaccine, consisting of Leishmania major cysteine proteinases Type I (CPB) and Type II (CPA), partially protects against leishmaniasis. AB - Cysteine proteinases (CPs) are enzymes that belong to the papain superfamily, which are found in a number of organisms from prokaryotes to mammals. On the parasitic protozoan Leishmania, extensive studies have shown that CPs are involved in parasite survival, replication and the onset of disease, and have, therefore, been considered as attractive drugs and/or vaccine targets for the control of leishmaniasis. We have previously shown that cysteine proteinases, Type I (CPB) and Type II (CPA), in Leishmania major (L. major), delivered as recombinant proteins or in plasmid DNA, induce partial protection against infection with the parasite in BALB/c mice. We had shown that the level of protection was greater if a cocktail of cpa and cpb containing DNA constructs was used. Therefore, to reduce the costs associated with the production of these vaccine candidates, a construct was developed, whereby the cpa and cpb genes were fused together to give rise to a single hybrid protein. The genes were fused in tandem where the C-terminal extension (CTE), encoding region of CPB, was located at the 3' of the fused genes, and ultimately expressed in the bacterial expression construct pET-23a. The expression of the CPA/B hybrid protein (60 kDa) was verified using rabbit anti-CPA and anti-CPB antibodies by SDS-PAGE and immunoblotting. The protective potential of the CPA/B hybrid protein against the infection with Leishmania was then assessed in BALB/c mice. The animals were vaccinated with CPA/B, challenged with live L. major promastigotes, and the degree of protection was examined by measuring footpad lesion sizes. It was found that there was a delay in the expansion of lesions size compared to control groups. Furthermore, an immunological analysis of antibody isotypes, before and after infection, showed high levels of IgG2a compared to IgG1 (more than five fold) in the CPA/B hybrid protein vaccinated group. In addition, a predominant Th1 immune response characterized by in vitro IFN-gamma production was observed, along with little, if any, IL-5 production. This finding indicates that the hybrid CPA/B is able to elicit a protective immune response against L. major in the mice model. In addition, 54% of individuals tested, who had recovered from cutaneous leishmaniasis, produced more than 50 pg/ml IFN-gamma, in response to the CPA/B hybrid protein in an in vitro assay, demonstrating the importance of cysteine proteinases as targets of immune response in humans. PMID- 15121306 TI - Immune responses of anti-HAV in children vaccinated with live attenuated and inactivated hepatitis A vaccines. AB - The immunogenicity of a live attenuated HAV vaccine and an inactivated HAV vaccine was compared. Altogether 117 children were vaccinated with either the inactivated or the live attenuated vaccine. Children were bled at months 1, 6, 7, 12 and 24, and the anti-HAV total IgG antibody and IgG subclass profile were assessed. In both vaccinated groups, the geometric mean titer (GMT) of anti-HAV peaked 7 months after the initial dose and declined during the following months. The IgG subclass profiles in both vaccinated groups were highly restricted to IgG1 and IgG3. Both vaccines have been shown highly effective in preventing viral hepatitis A in former studies. PMID- 15121308 TI - HIV risk and prevention in a post-vaccine context. AB - Initial HIV vaccines are likely to be only partially efficacious; increased risk behaviors in response to future HIV vaccine availability have the potential to subvert the effectiveness of vaccines in controlling the AIDS epidemic. To assess attitudes, beliefs and behavioral intentions in response to hypothetical availability of FDA-approved HIV vaccines, we conducted 9 focus groups among participants (N = 99; median age = 33 years; 48% female; 22% African American, 44% Latino, 28% White) recruited from STD clinics, needle exchange programs, and Latino community based health organizations, using purposive, venue-based sampling, and interviewed 9 key informant service providers. Data were analyzed using narrative thematic analysis and Ethnograph qualitative software. Participants predicted a "lightening up" of safer sex behaviors among at least half of their peers and, to a lesser extent, a relaxing of safer needle use practices in response to HIV vaccine availability. Both participants and providers urged HIV preventive interventions that: (1) provide education and awareness regarding partial efficacy vaccines, (2) combat the belief in an HIV vaccine as a "magic bullet," and (3) stressed the need for sustained behavioral risk reduction interventions in the face of continued HIV risk and other STDs. PMID- 15121307 TI - Glycol chitosan improves the efficacy of intranasally administrated replication defective human adenovirus type 5 expressing glycoprotein D of bovine herpesvirus 1. AB - The ability of two soluble formulations, namely chitosan and glycol chitosan, when used as an intranasal adjuvant, to improve the immunogenicity of an intranasal human adenovirus type 5 replication defective expressing bovine herpesvirus 1 (BoHV-1) glycoprotein D based vaccine, was investigated in cattle. Their adjuvant effects on immune response by increasing clinical and especially virological protection against an intranasal BoHV-1 challenge were then evaluated. The best virological protection was obtained in calves immunized with the vaccine vector adjuvanted with glycol chitosan which decreased the challenge BoHV-1 virus excretion titres by 0.5-1.5 log when compared to those obtained in calves immunized with the vaccine vector alone or adjuvanted with chitosan. A slight difference in clinical scores was observed in calves immunized with the adjuvanted vaccine vector compared to calves immunized with the vaccine vector alone. The obtained data suggest that the tested soluble formulation of glycol chitosan has promising potential use as an intranasal adjuvant for recombinant viral vector vaccines in cattle. PMID- 15121309 TI - A single vaccination with protein-microspheres elicits a strong CD8 T-cell mediated immune response against Mycobacterium tuberculosis antigen Mtb8.4. AB - Efficient protein-based vaccine delivery systems are needed to achieve a persistent memory immune response capable of detecting and eliminating intracellular pathogens such as Mycobacterium tuberculosis (TB). We have developed a novel protein-microsphere formulation using the recently discovered TB antigen Mtb8.4. Immunization of mice with a single dose of this Mtb8.4 microsphere formulation resulted in both humoral and cellular responses against Mtb8.4. The Mtb8.4-specific CD8 T-cell responses following a single administration of Mtb8.4-microspheres exceeded that elicited by protein plus adjuvant following multiple immunizations. These results demonstrate the efficacy of a single dose protein-microsphere vaccine for the induction of strong cell mediated and humoral immune responses against M. tuberculosis antigens. PMID- 15121310 TI - Distribution of adsorbed antigen in mono-valent and combination vaccines. AB - The distribution of alpha-casein, bovine serum albumin (BSA), myoglobin and recombinant protective antigen (rPA) in mono-valent and combination vaccines containing aluminum hydroxide adjuvant was studied by fluorescence microscopy and flow cytometry. Green and red fluorescent probes were conjugated to the antigens. Adsorption isotherms of the fluorescently labeled proteins to aluminum hydroxide adjuvant demonstrated that incorporation of the fluorescent probe did not significantly affect the adsorption. In mono-valent vaccine systems, antigen adsorption occurred within one minute and uniform surface coverage of the adjuvant aggregates was observed within 1h. Content uniformity was achieved through a cycle of de-aggregation and re-aggregation of the aluminum hydroxide adjuvant aggregates caused by mixing. For combination vaccines, two antigens were adsorbed separately to the aluminum hydroxide adjuvant prior to combination. Following combination, cycles of de-aggregation and re-aggregation occurred due to mixing, which led to uniform distribution of both antigens. The results of this study indicate that content uniformity should not be an issue during the production of mono-valent or combination vaccines as long as adequate mixing procedures are followed. PMID- 15121311 TI - Antennapedia transduction sequence promotes anti tumour immunity to epicutaneously administered CTL epitopes. AB - The identification of tumor antigens has spurred the development of efficient adjuvants and novel delivery systems for cancer immunotherapy. To this end, a peptide-based vaccine consisting of the Antennapedia transduction sequence (ANTP) attached to an antigenic peptide was designed to enhance per-cutaneous delivery into cells of the epidermis and dermis. Here we show that the topical application of OVA(257-264) linked to ANTP in mice onto tape-stripped skin resulted in enhanced delivery of the antigen through the skin whereas OVA(257-264) alone remained distributed uniformly on the skin surface. This delivery correlated with an increase in the CTL response against OVA. When mixed with CpG oligodinucleotides (ODN), the recombinant antigen protected mice from tumor challenge. These data provide the first indication that in vivo use of a translocation sequence can enhance delivery of therapeutic peptides and increase anti-tumor immunity through a simple and safe mechanism involving enhanced penetration of the skin barrier. PMID- 15121312 TI - The societal value of universal childhood vaccination. AB - Availability of new vaccines preventing infectious diseases in healthy children populations is increasing worldwide. In Canada, despite the current recommendation of the National Advisory Committee on Immunization to include recent vaccines in routine schedule, only a few provinces have incorporated some of the newer vaccines in routine vaccination programs. A review was undertaken of economic evaluations of childhood vaccination strategies performed from the societal point of view in industrialized countries, to gain perspective on their global benefits. The general trend supports most universal vaccination programs as cost-saving or cost-effective for society. Comparison of vaccination programs with other health care interventions indicates that vaccines are often one of society's best healthcare investments. Current data suggest that the Canadian society would benefit from a more complete immunization program. PMID- 15121313 TI - Clinical experience of the 23-valent capsular polysaccharide pneumococcal vaccination in HIV-1-infected patients receiving highly active antiretroviral therapy: a prospective observational study. AB - To assess the impact of vaccination with 23-valent pneumococcal polysaccharide vaccine on the risks for development of pneumococcal disease, all-cause community acquired pneumonia, HIV progression, and mortality and immunologic and virologic responses among HIV-1-infected patients treated with highly active antiretroviral therapy (HAART), we conducted a 2-year prospective observational cohort study at a university hospital in Taiwan. A total of 305 HIV-1-infected patients who received 23-valent pneumococcal vaccine (vaccinees) and 203 patients who did not (non-vaccinees) were prospectively observed between 1 June 2000 and 31 October 2002. Changes of CD4+ and plasma viral load (PVL) from baseline to week 4 of vaccination were assessed in 31 randomly selected vaccinees. The incidence of pneumococcal disease and bacteremia of vaccinees was 2.1 per 1000 patient-years (PY) (95% confidence interval (95% CI), 1.7-2.5 per 1000 PY) over the median observation of 641 days (range, 37-832 days) following vaccination while that of non-vaccinee was 21.8 per 1000 PY (95% CI, 20.1-23.7 per 1000 PY) and 7.3 per 1000 PY (95% CI, 7.0-7.6 per 1000 PY), respectively, over the observation of 500 days (range, 32-851 days), with an adjusted odds ratio (AOR) for developing pneumococcal disease of 0.085 (95% CI, 0.010-0.735) and for bacteremia of 0.22 (95% CI, 0.018-2.561). The median CD4+ count increased by 45 x 10(6) l(-1) (P = 0.01) and median PVL change was 0 log(10) copies/ml (range of decrease, -0.74 to 2.47 log(10) copies/ml) after 1 month of pneumococcal vaccination among the subgroup of 31 vaccinees receiving HAART. The median CD4+ count increase from baseline to the end of study was 149 x 10(6) l(-1) for vaccinees and 107 x 10(6) l(-1) for non-vaccinees (P = 0.21). The AOR of developing all-cause community acquired pneumonia and new AIDS-defining opportunistic illnesses (OI) of vaccinees as compared to non-vaccinees was 1.876 (95% CI, 0.785-4.485) and 0.567 (95% CI, 0.217-1.484), respectively. Death rate of vaccinees and non-vaccinees was 17.7 per 1000 PY (95% CI, 16.5-18.9 per 1000 PY) and 80.5 per 1000 PY (95% CI, 77.1-83.9 per 1000 PY), respectively. Adjusted hazard ratio for death of vaccinees as compared with non-vaccinees was 0.733 (95% CI, 0.236-2.274). Our data suggested that vaccination with 23-valent pneumococcal polysaccharide vaccine and receipt of HAART were associated with reduced risks for pneumococcal disease among HIV-1-infected patients receiving HAART. Vaccination did not increase the risks of all-cause community-acquired pneumonia, HIV progression, and mortality. Vaccination did not increase PVL or decrease CD4+ among HIV-1 infected patients receiving HAART. PMID- 15121314 TI - Hospitalisations for gastroenteritis: the role of rotavirus. AB - To determine the proportion of hospitalisations for gastroenteritis caused by rotavirus, we tested for rotavirus stool samples of all children under the age of five hospitalised for gastroenteritis between 1 December 1999 and 30 May 2000 in seven community and specialised hospitals in Quebec. Of 944 children hospitalised, 565 (59.9%) were screened for rotavirus and 405 (71.7%) tested positive. From December to April, the proportion of positive results rose from 51.6 to 78.1%. Compared with children whose test results were negative, children who tested positive presented vomiting more frequently upon admission (88.9 versus 60.4%) and needed IV fluids in greater proportion (94.1 versus 78.0%), but spent less time in hospital (2.8 versus 3.3 days). Aside from dehydration, no complications were noted. In Quebec, a large majority of winter and spring hospitalisations for gastroenteritis in children is attributable to rotavirus. PMID- 15121315 TI - Catch-up vaccination against Haemophilus influenzae type b in human immunodeficiency virus-infected Thai children older than 2 years old. AB - Although most of Thai children older than 2 years are immune against Haemophilus influenzae type b (Hib) without prior vaccination, it may not be the case in HIV infected children. Of 44 HIV-infected children tested before vaccination at the mean age of 36 months (range 24-84 months), 32 (73%) were susceptible (anti-PRP <0.15 microg/ml). At 6 months after a single dose of tetanus-conjugated Hib vaccination, 67% developed anti-PRP >/=0.15 microg/ml, however, only 33% developed titer of >/=1 microg/ml. Four of seven (57%) with anti-PRP 0.15-0.99 microg/ml at baseline were boosted to the titer of >/=1 microg/ml after vaccination. Seroconversion rate and geometric mean titer (GMT) level in response to the vaccination did not correlate with HIV stage, but did correlate with viral load level of 100,000 copies/ml. HIV-infected children older than 2 years would benefit from Hib vaccination, although, one dose catch-up schedule is not sufficient in a third of these children. A second dose is needed in these children especially those with viral load of level of >100,000 copies/ml. PMID- 15121316 TI - Immunogenicity of Plasmodium vivax merozoite surface protein-9 recombinant proteins expressed in E. coli. AB - Merozoite surface protein-9 of Plasmodium vivax (PvMSP-9) is highly conserved and present in several malaria species. Here, we present the immunogenic properties of two recombinant glutathione S-transferase (GST) fusion proteins comprising the N-terminus (PvMSP-9-Nt) and the second block of tandem repeats (PvMSP-9-RepII) of PvMSP9. These recombinants proteins were used to immunize BALB/c mice. The specificity and subtyping of the antibodies and the cellular immune responses were evaluated by enzyme-linked immunosorbent assay (ELISA) and ELISPOT, respectively, using the recombinant proteins as antigens. Our results demonstrate that both the N-terminal and the tandem repeat regions of MSP9 are immunogenic in mice. The ELISA antibody titers elicited by PvMSP-9-Nt were significantly higher (1:819,200) than the antibody titers elicited by PvMSP-9-RII (1:409,600). Analysis of IgG subclasses showed that both recombinant proteins induce similar antibody patterns where IgG1, IgG2a and IgG2b were most predominant. Moreover, all sera from mice immunized with either PvMSP-9-Nt or PvMSP-9-RII, which were positive by ELISA showed reactivity with P. vivax, P. cynomolgi, P. knowlesi and P. coatneyi schizonts by immunofluorescence assays (IFA). Similar results were observed in western immunoblot analyses using parasite extracts. Furthermore, immunization of mice with the PvMSP-9-Nt upon stimulation with PvMSP-9-Nt secreted IFN-gamma and IL-5. We have also used the two PvMSP-9 recombinant constructs to show that individuals exposed to P. vivax infections in an endemic area of Brazil had IgG antibodies reactive with the recombinant proteins. PMID- 15121319 TI - The Cochrane Vaccines Field. AB - The Cochrane Vaccines Field (CVF) is a Cochrane Collaboration entity. The CVF was founded to facilitate the gathering of evidence on vaccines and their effects (ability to prevent the target disease, short and long term safety and efficiency) finalised to the production of systematic reviews. The CVF formulate criteria for population-based quality assessment of vaccines. The Cochrane Vaccines Field was registered in 1996 and is now based in Alessandria, Italy since 2000. The specific contribution of the Cochrane Vaccines Field is to promote the preparations and maintenance of systematic reviews on the effects of vaccines facilitating the coverage of the knowledge gaps in vaccinology and thus forming a methodological focus for a population based assessment of vaccines and systematic gathering and dissemination of information on vaccines. The targets reached until now are: the handsearch of Vaccine and The Journal of Medical Virology, the updating of an overview on the current status of knowledge on vaccines, the preparation and maintenance of systematic reviews. The next Key objective of the Cochrane Vaccines Field is to develop "The Vaccines Register" a register comprising all relevant studies on the effectiveness, the safety and the economical aspects of vaccines. This Register aims to be an extensive and up to date reference to enable those making decisions on the use of human vaccines to make best use of available evidence assessed by its reliability. PMID- 15121317 TI - Humoral immunoreaction induced by TCR DNA vaccine for beta chain of T cell lymphoma. AB - We exploited the humoral immunoreaction of mice induced by TCR DNA vaccine of beta chain of T cell lymphoma. The plasmids of pcDNA3.1/TCR V beta 8 was constructed. The BALB/c mice were randomly divided into four groups which were pcDNA3.1, pcDNA3.1/TCR V beta 8, pcDNA3.1/TCR V beta 8 + CpG + liposome and phosphorothioate CpG groups with six mice in each group. Vaccines were injected in bilateral musculus quadriceps femoris of mice in the 0, second, and fourth week, respectively. The antibody formation was tested by indirect immuofluorescence in the 0, second, fourth, sixth and eighth weeks, respectively, before and after immunization. Production of antibody against TCR V beta 8 antigen was observed in the groups of pcDNA3.1/TCR V beta 8 and pcDNA3.1/TCR V beta 8 + CpG + liposome. The antibody titer began to rise in the fourth week and attain the maximal value in the sixth week. The antibody titer in the group of pcDNA3.1/TCR V beta 8 + CpG + liposome was higher than that in the group of pcDNA3.1/TCR V beta 8 in the fourth and eighth weeks (both P<0.01); the antibody titer in the group of pcDNA3.1/TCR V beta 8 + CpG + liposome was markedly higher than that in the group of pcDNA3.1/TCR V beta 8 in the sixth week (P<0.001). The result indicate that TCR V beta 8 antigen can induce formation of special antibody in mice. CpG and liposome can improve the humoral immunoreaction induced by TCR V beta 8 gene vaccine. PMID- 15121320 TI - The Brighton Collaboration-enhancing vaccine safety. PMID- 15121321 TI - Evaluation of vaccine safety after the events of 11 September 2001: role of cohort and case-control studies. AB - As immunization programs world-wide "mature" with high vaccine coverage and near elimination of vaccine-preventable disease, vaccine safety issues have increased in relative prominence. In the wake of events of 11 September 2001, fear of bioterrorism has reemerged. The paradigm of eradicating vaccine-preventable diseases, stopping vaccinations and thereby also eradicating the associated vaccine adverse events (a la smallpox) may unfortunately be obsolete. If all vaccinations have to be continued indefinitely, research is needed more than ever to understand and prevent rare vaccine adverse events. Case-control studies are usually best suited for such purposes, especially when nested within a pre existing large-linked administrative database cohort to minimize bias. The new clinical immunization safety assessment centers may play an important role in bridging the sometimes conflicting clinical and epidemiologic perspectives in vaccine safety. PMID- 15121322 TI - Technologies that make administration of vaccines safer. AB - There is an ever-expanding technology that is aimed at making the administration of vaccines safer. Conventional ways of administering vaccines are being upgraded, modified or replaced by a wide variety of innovations. The paradigm of liquid vaccines, needles and syringes is slow to change, but already developments are occurring that will change for ever the way vaccines are administered and will improve the safety record of immunization. The oral route of vaccine administration has generally been thought of as safe, but until now only the polio vaccine has been widely used in this way. The conventional method of vaccine administration is by injection. Many ingenious devices have become available that now make injecting safer. Inventors are now looking imaginatively to alternative routes and technologies for delivering vaccines. Vaccines are generally manufactured to extremely high standards and rarely are shown to be the cause of safety issues. People remain the weakest safety link is vaccine administration. Technologies that bypasses the ability of man to make bad decisions or to behave incorrectly are of tremendous value. The vaccine world is in the middle of a radical re-think about how vaccines might best be administered. The presentation of the vaccine can be altered to fit new technologies such as powder jet guns, or skin patches. Even the conventional needle and syringe have evolved to much safer versions, and are set to continue this evolution. All this means even safer vaccines and their delivery. PMID- 15121323 TI - A global perspective on vaccine safety. AB - "Immunization Safety" (i.e., ensuring and monitoring the safety of all aspects of immunization, including vaccine quality, storage and handling, vaccine administration and the disposal of sharps) remains a major challenge. Any vaccine safety issue, real or perceived, may lead to rumors and undermine confidence in vaccination and, ultimately, have dramatic consequences for immunization coverage and disease incidence. In 1999, WHO's Department of Vaccines and Biologicals launched the Immunization Safety Priority Project to establish a comprehensive system to ensure the safety of all immunizations given in national immunization programmes. The project aims at strengthening each country's capacity. The principal areas of activity of the project include: research and development of safer vaccines and safer and simpler vaccine delivery technologies; assurance of vaccine safety through quality control procedures and quality specifications; implementation of tools to ensure vaccine quality up to vaccine administration; access to safe and efficient vaccine administration technologies and their disposal; and identification and management of risks related to immunization. This report focuses on the latter area and highlights challenges and critical factors in establishing a safety profile for a vaccine and the importance and limits of post market surveillance. It presents some of WHO's supporting activities among which the establishment of a Global Advisory Committee on Vaccine Safety to provide an independent scientific assessment of vaccine safety issues. WHO has a role to play not only because of its technical and normative role but also because of its privileged relation with country authorities and its global vision and mandate and being perceived as neutral and free of conflicts of interest. PMID- 15121324 TI - Control without separate controls: evaluation of vaccine safety using case-only methods. AB - Epidemiological methods involving only cases are reviewed in the context of vaccine safety studies. These methods include some ecological methods, case coverage methods, case-crossover and self-controlled case series methods. The properties of the methods are described using examples from the literature. It is argued that such methods, and in particular the self-controlled case series method, are powerful epidemiologic tools meriting the same attention as more traditional cohort and case-control methods. PMID- 15121325 TI - The success of immunization-shovelling its own grave? AB - Successful immunization programmes have reduced the burden of a number of infectious diseases on a global scale. Yet, as the fear of sequelae of vaccine preventable diseases diminishes in the public, the focus of interest has shifted towards true and alleged "side effects". Maintaining confidence in the necessity, tolerability and safety of immunizations is of paramount importance today. This requires, amongst other prerequisites, precise definitions of "adverse events following immunisation". In Europe, a collaborative effort named EUSAFEVAC in concert with the globally active "Brighton Collaboration" has been initiated. Volunteers from academic institutions, vaccine licensing authorities, public health institutes, governmental organizations, safety units within the vaccine manufacturing industry as well as practicing physicians are working together to achieve this goal. PMID- 15121326 TI - The role of editorial peer review in the evaluation of vaccine safety. PMID- 15121327 TI - Use of vaccine surveillance data in the evaluation of safety of vaccines. AB - In this article we discuss some of the most widely used methods of the routine vaccine safety surveillance. The main problems inherently associated with these passive techniques are also addressed. The vaccine safety surveillance in Finland is used as an example. Finally, we are giving a brief introduction to more advanced surveillance tools, namely record linkage and electronic vaccine registers, which are now being investigated in Finland as a part of the European Research Programme for Improved Vaccine Safety Surveillance (EUSAFEVAC)-project. PMID- 15121328 TI - Methodological issues arising from systematic reviews of the evidence of safety of vaccines. AB - Adaptations to the recognized methods of systematic reviewing are required when addressing questions about safety, particularly about rare and/or long-term serious adverse events. Conducting a systematic review of vaccine safety requires the implementation of novel strategies for locating studies, the use of experimental instruments to assess the quality of non-randomized studies, and the employment of pooling methods for non-randomized data, where appropriate. Standardizing both the indexing of adverse event data on electronic libraries and their reporting would improve the potential of systematic reviews of vaccine to draw accurate conclusions about the safety of a vaccine. PMID- 15121329 TI - Ethical aspects of the methods used to evaluate the safety of vaccines. PMID- 15121330 TI - Field evaluation of vaccine safety. AB - Surveillance of adverse events in the field represents an essential part of vaccination strategies. Safety studies conducted before vaccine licensure have often not enough power to detect rare or long-term adverse events. Furthermore, adverse events may be caused by incorrect procedures of storage or administration. Conducting surveillance of adverse events in the field is necessary for detecting unexpected increases in incidence of adverse events, for continuing education of health personnel involved in vaccine programs, for providing appropriate information to the public, and for supporting interventions to prevent further adverse events. Signals generated by surveillance systems may stimulate to conduct formal studies to assess causality. PMID- 15121331 TI - Effects of green tea polyphenol on preservation of human saphenous vein. AB - The potential role of green tea polyphenol (GtPP) in preserving the human saphenous vein was investigated under physiological conditions. The vein segments were incubated for 1, 3, 5, 7 and 14 days, either after 4h of treatment with 1.0mg/ml GtPP or in the presence of GtPP at the same concentration. After incubation, the endothelial cell viability, endothelial nitric oxide synthase (eNOS) expression and the vein histology were evaluated. When the veins were not treated with GtPP, the viability of the endothelial cells was significantly reduced with the progress in the culture time, and none of the cells expressed eNOS after 5 days. Furthermore, severe histological changes and structural damage were observed in the non-treated veins. In contrast, incubating the veins after 4h of GtPP treatment significantly prevented these phenomena. The cellular viability of the GtPP-treated vein was approximately 64% after 7 days, and eNOS expression was maintained up to 40%, compared to that of the fresh vein. The histological observations showed that the vasculature was quite similar to that of the fresh vein. When incubated with GtPP, the vein could also be preserved for 1 week under physiological conditions retaining both its cellular viability (61%) and eNOS expression level (45%) and maintaining its venous structure without any morphological changes. These results demonstrate that GtPP treatment may be a useful method for preserving the HSV. PMID- 15121332 TI - Enhanced process monitoring of fed-batch penicillin cultivation using time varying and multivariate statistical analysis. AB - On-line monitoring of penicillin cultivation processes is crucial to the safe production of high-quality products. In the past, multiway principal component analysis (MPCA), a multivariate projection method, has been widely used to monitor batch and fed-batch processes. However, when MPCA is used for on-line batch monitoring, the future behavior of each new batch must be inferred up to the end of the batch operation at each time and the batch lengths must be equalized. This represents a major shortcoming because predicting the future observations without considering the dynamic relationships may distort the data information, leading to false alarms. In this paper, a new statistical batch monitoring approach based on variable-wise unfolding and time-varying score covariance structures is proposed in order to overcome the drawbacks of conventional MPCA and obtain better monitoring performance. The proposed method does not require prediction of the future values while the dynamic relations of data are preserved by using time-varying score covariance structures, and can be used to monitor batch processes in which the batch length varies. The proposed method was used to detect and identify faults in the fed-batch penicillin cultivation process, for four different fault scenarios. The simulation results clearly demonstrate the power and advantages of the proposed method in comparison to MPCA. PMID- 15121333 TI - Rational affinity purification of native Streptomyces family 10 xylanase. AB - Xylanase SoXyn10A from Streptomyces olivaceoviridis E-86 comprises a family 10 catalytic module linked to a family 13 carbohydrate-binding module (SoCBM13). The SoCBM13 has a beta-trefoil structure, with binding sites in each subdomain (alpha, beta and gamma). Subdomain alpha, but not subdomains beta and gamma, binds tightly to lactose. It was, therefore, thought that immobilized lactose could be used for the affinity purification of SoXyn10A. Lactosyl-Sepharose was prepared and tested as an affinity matrix. SoXyn10A produced from the cloned xyn10A gene by Escherichia coli, and native SoXyn10A in culture supernatants from S. olivaceoviridis, were purified to homogeneity in a single step by affinity chromatography using this matrix. This simple purification of SoXyn10A makes the enzyme an attractive candidate for applications requiring xylanase. The CBM also has the potential for use as an affinity tag for the purification of other proteins. PMID- 15121334 TI - Effect of pH on metabolic pathway shift in fermentation of xylose by Clostridium tyrobutyricum. AB - The effect of pH (between 5.0 and 6.3) on butyric acid fermentation of xylose by Clostridium tyrobutyricum was studied. At pH 6.3, the fermentation gave a high butyrate production of 57.9 g l(-1) with a yield of 0.38-0.59 g g(-1) xylose and a reactor productivity up to 3.19 g l(-1)h(-1). However, at low pHs (<5.7), the fermentation produced more acetate and lactate as the main products, with only a small amount of butyric acid. The metabolic shift from butyrate formation to lactate and acetate formation in the fermentation was found to be associated with changes in the activities of several key enzymes. The activities of phosphotransbutyrylase (PTB), which is the key enzyme controlling butyrate formation, and NAD-independent lactate dehydrogenase (iLDH), which catalyzes the conversion of lactate to pyruvate, were higher in cells producing mainly butyrate at pH 6.3. In contrast, cells at pH 5.0 had higher activities of phosphotransacetylase (PTA), which is the key enzyme controlling acetate formation, and lactate dehydrogenase (LDH), which catalyzes the conversion of pyruvate to lactate. Also, PTA was very sensitive to the inhibition by butyric acid. Difference in the specific metabolic rate of xylose at different pHs suggests that the balance in NADH is a key in controlling the metabolic pathway used by the cells in the fermentation. PMID- 15121335 TI - Stability of the ANAMMOX process in a gas-lift reactor and a SBR. AB - In the last years, the ANAerobic AMMonium OXidation (ANAMMOX) process has been put forward as a promising alternative to treat ammonium rich wastewaters. An ANAMMOX gas-lift reactor and a sequential batch reactor (SBR) were operated during around 200 days in this study, reaching nitrogen loading rates (NLRs) of 2.0 and 0.75 g l(-1) per day, respectively. The efficiency in the nitrite (limiting substrate) removal was 99%. The ammonium and nitrite influent concentrations were increased stepwise until biomass in the reactors started to float. These flotation events coincided with periods when the NLR exceeded the maximum specific ANAMMOX activity (MSAA) of the sludge. The MSAA, determined in batch experiments, was 0.9 and 0.44 g g(-1) per day for biomasses from the gas lift reactor and the SBR, respectively. Flotation of the biomass occurred most likely due to a granule density decrease caused by dinitrogen gas accumulation inside the granules and an apparent breakage of the granules. Further research is needed to understand this phenomenon and to optimise the corresponding strategies to counteract the flotation. PMID- 15121336 TI - Strategies for fed-batch cultivation of t-PA producing CHO cells: substitution of glucose and glutamine and rational design of culture medium. AB - A strategy for fed-batch cultivation of t-PA producing recombinant CHO cells is presented, based on the substitution of glucose and glutamine for slowly metabolized nutrients and in a rational design of the medium. Media for the batch and fed stages were based on the cell specific amino acid requirements, which allowed a more accurate determination of the initiation of the fed stage and the frequency of nutrient addition from then on. Salt concentration was also reduced in both media to avoid an increase in osmolality. As a consequence of this rational design, most amino acid did not accumulate significantly during the fed stage, as usually occurs when their supply is not based on cell requirements; also, lower amounts of by-products were obtained when osmolality level was kept low, that altogether increased viability, longevity and t-PA production when compared with a reference batch culture. Alternating glucose and galactose during the fed stage, allowed lactate detoxification of the cells through their own metabolism. This allowed an increase in cell growth and cell viability with respect to a fed-batch culture in which only glucose was used in the fed stage. PMID- 15121337 TI - Modeling and experimental validation of the signal transduction via the Escherichia coli sucrose phospho transferase system. AB - Bacterial signal processing was investigated concerning the sucrose phosphotransferase system (sucrose PTS) in the bacterium Escherichia coli as an example. The about 20 different phosphotransferase systems (PTSs) of the cell fulfill besides the transport of various carbohydrates, also the function of one signal processing system. Extra- and intracellular signals are converted within the PTS protein chain to important regulatory signals affecting, e.g. carbon metabolism and chemotaxis. A detailed dynamical model of the sucrose PTS was developed describing transport and signal processing function. It was formulated using a detailed description of complex formation and phosphate transfer between the chain proteins. Model parameters were taken from literature or were identified with own experiments. Simulation studies together with experimental hints showed that the dynamic behavior of phosphate transfer in the PTS runs within 1 s. Therefore a description of steady state characteristics is sufficient for describing the signaling properties of the sucrose PTS. A steady state characteristic field describes the degree of phosphorylation of the PTS protein EIIACrr as a function of the input variables extracellular sucrose concentration and intracellular phosphoenolpyruvate (PEP):pyruvate ratio. The model has been validated with different experiments performed in a CSTR using a sucrose positive E. coli W3110 derivative. A method for determining intracellular metabolite concentrations has been developed. A sample preparation technique using a boiling ethanol buffer solution was successfully applied. The PTS output signal degree of phosphorylation of EIIACrr was also measured. Steady state conditions with varying dilution rate and dissolved oxygen concentration and dynamical variations applying different stimuli to the culture were considered. Pulse, and stop feeding experiments with limiting sucrose concentrations were performed. Simulation and experimental results matched well. The same holds for the expanded sucrose PTS and glycolysis model. PMID- 15121338 TI - Prevention of interfacial inactivation of enzymes by coating the enzyme surface with dextran-aldehyde. AB - Interactions between soluble enzymes and interfaces of organic solvent drops or gas bubbles have a very negative effect on the operational stability of the soluble enzymes. In this study, the formation of a hydrophilic shell around the enzyme has been attempted using dextran-aldehyde which would prevent the interaction between enzyme and hydrophobic interfaces with minimal modification of the enzyme surface. After optimizing the size of the dextran (that was found to play a critical role), three different enzymes (glucose oxidase, d-amino acid oxidase, and trypsin) have been conjugated with dextran-aldehyde and their stability towards organic-aqueous and air-liquid interfaces has been evaluated. The treatment itself proved to be very low-cost in terms of activity and was highly stabilizing for the three enzymes assayed. The conjugated preparation of the three assayed enzymes remained fully active in the presence of air-liquid interfaces for at least 10h. However, the unmodified enzymes lost more than 50% of activity within the first hour of the experiments except for trypsin which kept 38% activity after 12h while the trypsin dextran-aldehyde conjugate maintained 100% enzyme activity. Similar results were achieved in the presence of stirred organic solvent-aqueous buffer biphasic system, although in this case some activity was lost by the action of the soluble portion of the organic solvent. In fact, this treatment seems to be also effective to improve the resistance to the action of organic solvent. PMID- 15121339 TI - Integration of purification with immobilization of Candida rugosa lipase for kinetic resolution of racemic ketoprofen. AB - The two processes for the partial purification and for the immobilization of a crude lipase preparation (Candida rugosa Lipase OF) have been successfully integrated into one by simple adsorption of the enzyme onto a cation ion exchanger resin (SP-Sephadex C-50) at pH 3.5. Due to selective removal of the unfavorable lipase isoenzyme (L1), the enzyme components (mainly L2 and L3) that are tightly fixed on the resin displayed a significantly improved enantioselectivity (E value: 50 versus 13 with addition of Tween-80) in the biocatalytic hydrolysis of 2-chloroethyl ester of rac-ketoprofen. The activity yields of the immobilized lipase were 48 and 70%, respectively when emulsified and non-emulsified substrates were employed for enzyme assay. Moreover, the concentration of Tween-80 was found to be a factor affecting the lipase enantioselectivity. By using such an immobilized enzyme as biocatalyst, the process for preparing enantiopure (S)-ketoprofen becomes simpler and more practical as compared with the previously reported procedures and the product was obtained with >94% ee at 22.3% conversion in the presence of an optimal concentration (0.5 mg/ml) of Tween-80 at pH 3.5. Furthermore, the operational stability of the immobilized biocatalyst was examined in different types of reactors. In an air-bubbled column reactor, the productivity was much higher than that in a packed-bed column reactor, in spite of a slightly lower stability. Under optimal conditions, the air-bubbled column reactor could be operated smoothly for at least 350 h, remaining nearly 50% activity. PMID- 15121341 TI - The many forms of bipolar disorder: a modern look at an old illness. AB - Bipolar disorder continues to be underrecognized, despite being known for 2000 years. Mania, the fullest expression of the disease affects approximately 1% of the population; the less-than-manic forms of the disease dominated by depressive episodes have recently been found to be more common, affecting 4-5% of the population. In reviewing the international literature on this broadened bipolar spectrum, this paper pays particular tribute to the French EPIDEP and EPIMAN studies and Italo-American collaboration which have generated the largest set of systematic data on the new clinical portrait of bipolar disorders. Early detection is crucial, because untreated bipolar disorder has a high mortality rate. A review of the diagnostic criteria for the various subtypes of bipolar disorder has identified several factors that interfere with making an accurate diagnosis. These include age at onset, ethnic differences, co-morbidity (particularly substance abuse and alcoholism), and the broad range of clinical presentations. Moreover, symptoms frequently overlap with those of other psychiatric disorders including schizophrenia, attention-deficit disorder and personality disorders. Misdiagnosis is a major factor leading to a poor outcome for patients. Accurate identification and diagnosis of the different forms of mania can lead to specific treatment choices that may improve prognosis. Particularly important are recent data indicating reduced mortality with a variety of psychopharmacologic agents including, but not limited to, lithium and valproate. PMID- 15121342 TI - The effectiveness of divalproate in all forms of mania and the broader bipolar spectrum: many questions, few answers. AB - Divalproate (VPA) was the first drug, other than lithium, reported to be efficacious for the treatment of bipolar disorder. Since then, the effectiveness of VPA, alone, in combination, and as maintenance therapy, has been investigated in a number of studies. As a monotherapy, clinical studies revealed that VPA was superior to lithium with regard to being effective in a broader patient population: patients with depressive symptoms concurrent with their mania, patients previously poorly responsive to lithium, patients with more lifetime episodes, and on specific symptoms of elation/grandiosity and reduced need for sleep. In addition, while VPA had superior efficacy versus carbamazepine (CBZ), an anti-manic agent, and equivalent efficacy to the anti-psychotic agent olanzapine (OLZ) in manic patients with psychotic manic features, patients showed much better tolerability to VPA than CBZ, anti-psychotics, and, in particular, lithium. Recently, combination therapy (mood stabilizers plus anti-psychotic agents) has shown some advantages compared with monotherapy. Manic symptoms have been observed to improve to a greater extent when the mood stabilizer was co administered with an anti-psychotic rather than either agent used alone. While only a few maintenance studies have been conducted to date, two randomized studies have indicated consistent trends toward greater efficacy of VPA than lithium, and significant superiority over placebo on most, although not all, measures. In general, VPA provides a well-tolerated treatment that is efficacious for a broad spectrum of manic states and improves outcomes during maintenance treatment. PMID- 15121343 TI - Mood stabilizers: protecting the mood...protecting the brain. AB - The mechanism underlying the therapeutic action of mood stabilizers in bipolar disorder is not completely understood. The discovery that anticonvulsant agents, such as valproate (VPA), were effective in the treatment of bipolar disorder suggested a common biochemical mechanism(s) with lithium. Recent research has focused on how VPA and lithium change the activities of cellular signal transduction systems, especially the cyclic AMP and phosphoinositide second messenger pathways. Despite being structurally dissimilar, VPA produces effects on the protein kinase C (PKC) signalling pathway that are similar to lithium, although the VPA effects appear to be largely independent of myo-inositol. Furthermore, the therapeutic benefit of either drug require a prolonged administration suggesting alterations at the genomic level. Studies have revealed that both VPA and lithium altered the expression of several early inducible genes belonging to the AP-1 family of transcription factors; this family is responsible for controlling the expression of a number of genes including cytoprotective proteins such as the anti-apoptotic protein, bcl-2. Evidence shows that chronic administration of VPA or lithium can stimulate bcl-2 expression as well as inhibit GSK-3 beta activity, which renders a cell less susceptible to apoptosis. Thus, the mood stabilizers may act to restore the balance among aberrant signalling pathways in specific areas of the brain and prevent degeneration. PMID- 15121344 TI - Mental health visits to complementary and alternative medicine providers. AB - The objective of this study was to examine the characteristics of mental health visits to Complementary and Alternative Medicine (CAM) providers. A representative sample of acupuncturists, chiropractors, massage therapists, and naturopathic physicians in four states reported on 8933 consecutive visits, including demographic characteristics; presenting complaints; referral source; treatments provided; disposition; and other sources of care for the presenting problem. The proportion of visits for a mental health complaint ranged from 7% to 11% for acupuncture, massage, and naturopathic physicians to less than 1% for chiropractors. For acupuncturists, massage therapists, and naturopaths, 69-87% of patients making mental health visits were self-referred. The CAM provider discussed care with a conventional medical provider in 6-20% of cases and was aware of concomitant conventional medical care in an additional 10-30%. Only 1-5% were subsequently referred to conventional providers. For acupuncturists, massage therapists, and naturopaths, the proportion of visits for mental health concerns is similar to that in conventional primary care. Mental health visits to chiropractors are much less common, but this may reflect differences in true prevalence or differences in presentation. Among those seeking CAM care for mental disorders, concomitant treatment by conventional medical providers is common, but communication or coordination of care is rare. PMID- 15121345 TI - Relationship of sexual assault history to somatic symptoms and health anxiety in women. AB - Prior reports have pointed to a link between traumatic experiences and health consequences in women. The objective of this study was to determine whether there is an association between sexual assault history and measures of somatic symptoms and illness attitudes in a sample of female Veterans Affairs primary care patients, a group in whom high rates of sexual trauma have been reported. We conducted a cross-sectional study of a representative sample of 219 women in a Veteran's Affairs primary care outpatient clinic. Sexual assault history, somatic symptoms and health anxiety were assessed by self-report questionnaire. Multivariate analyses were used to examine relationships between sexual assault exposure and these outcomes. Ninety-seven women (43.9%) reported experience(s) of sexual assault (i.e., rape, attempted rape or being made to perform any type of sexual act through force or threat of harm). Sexual assault was associated with a significant increase in somatization scores, physical complaints across multiple symptom domains and health anxiety. Sexual assault was also a significant statistical predictor of having multiple sick days in the prior 6 months and of being a high utilizer of primary care visits in the prior 6 months. These data confirm a strong association between sexual trauma exposure and somatic symptoms, illness attitudes and healthcare utilization in women. Causal mechanisms cannot be inferred from these data. Studies in other cohorts are warranted. PMID- 15121346 TI - Patients' preferences in the treatment of depressive disorder in primary care. AB - Patients' preferences in the treatment of depression are important in clinical practice and in research. Antidepressant medication is often prescribed, but adherence is low. This may be caused by patients preferring psychotherapy, which is often not available in primary care. In randomized clinical trials, patients' preferences may affect the external validity. The aim of this article is to study patients' preferences regarding psychotherapy and antidepressant medication and the impact of these preferences on treatment outcome. A systematic review of the literature was performed. The majority of patients preferred psychotherapy in all available studies. Antidepressants were often regarded as addictive and psychotherapy was assumed to solve the cause of depression. Discussing and supporting preferences as part of a quality improvement program of depression care, resulted in more patients receiving the treatment that was most suitable to them. In two patient-preference trials, preferences did not influence treatment outcome. It can be concluded that a substantial percentage of well-informed patients prefer psychotherapy. Patients with strong preferences, mostly for psychotherapy, are likely not to enter antidepressant treatment or randomized clinical trials if their preferences are not supported. PMID- 15121347 TI - Use of brief depression screening tools in primary care: consideration of heterogeneity in performance in different patient groups. AB - Heterogeneity of performance of screening tools in different patient groups has rarely been considered in the literature on depression screening in primary care. The objectives of the present study were to assess and to compare diagnostic accuracy of three screening questionnaires (Brief Patient Health Questionnaire, General Health Questionnaire-12, WHO-5) in identifying depression across various patient subpopulations and to assess the accuracy of the unaided clinical assessment of primary care physicians in the same subgroups. We conducted a cross sectional validation study in 448 primary care patients. Two-by-two tables as well as receiver operating characteristics were applied. Results indicated that diagnostic accuracy (sensitivity, specificity) of the three screening instruments as well as of the clinical diagnoses differed in the various patient groups. Superiority of one screening tool over the other depends on the subgroup considered. Gender, age, form (subtype), and severity of depression influence the test characteristics of a screening tool. This should be considered if routine depression screening should be widely introduced. Of course, the benefit of routine screening also depends on efforts made for treatment and monitoring of patients in whom depression was diagnosed. PMID- 15121348 TI - The impact of a pharmacist intervention on 6-month outcomes in depressed primary care patients. AB - The object of the study was to evaluate outcomes of a randomized clinical trial (RCT) of a pharmacist intervention for depressed patients in primary care (PC). We report antidepressant (AD) use and depression severity outcomes at 6-months. The RCT was conducted between 1998 and 2000 in 9 eastern Massachusetts PC practices. We studied 533 patients with major depression and/or dysthymia as determined by a screening test done at the time of a routine PC office visit. The majority of participants had recurrent depressive episodes (63.5% with >/=4 lifetime episodes), and 49.5% were taking AD medications at enrollment. Consultation in person and by telephone was performed by a clinical pharmacist who assisted the primary care practitioner (PCP) and patient in medication choice, dose, and regimen, in accordance with AHCPR depression guidelines. Six month AD use rates for intervention patients exceeded controls (57.5% vs. 46.2%, P =.03). Furthermore, the intervention was effective in improving AD use rates for patients not on ADs at enrollment (32.3% vs. 10.9%, P =.001). The pharmacist intervention proved equally effective in subgroups traditionally considered difficult to treat: those with chronic depression and dysthymia. Patients taking ADs had better modified Beck Depression Inventory (mBDI) outcomes than patients not taking ADs, (-6.3 points change, vs. -2.8, P =.01) but the outcome differences between intervention and control patients were not statistically significant (17.7 BDI points vs. 19.4 BDI points, P =.16). Pharmacists significantly improved rates of AD use in PC patients, especially for those not on ADs at enrollment, but outcome differences were too small to be statistically significant. Difficult-to-treat subgroups may benefit from pharmacists' care. PMID- 15121350 TI - Prevalence and identification of alcohol use disorders among nonpsychiatric inpatients in one general hospital. AB - Alcohol use disorders (AUDs) are common among inpatients in general hospitals and often cause excess mortality. This study investigates the prevalence of AUDs among nonpsychiatric inpatients in one general hospital and evaluates the ability of medical staff to identify such morbidity. A two-phase case-identification strategy was employed utilizing the Alcohol Use Disorders Identification Test as the first-phase screening tool and the Schedules for Clinical Assessment in Neuropsychiatry as the second-phase diagnostic interview. Among 538 eligible patients, a total of 422 (78.4%) completed the first-phase screening. A subsample (20%) of those screened negative and 90% of those screened positive were interviewed at the second phase. The weighted 1-year prevalence rates of alcohol abuse and alcohol dependence were 3.9% and 12.6%, respectively. The overall identification rate of AUDs by medical staff was 25.4% (0% for alcohol abuse and 30% for alcohol dependence). In conclusion, approximately one sixth of nonpsychiatric inpatients in a general hospital have AUDs and have been neglected substantially by medical staff. Implications of the findings for the prevention of AUDs and their physical complications are discussed. PMID- 15121349 TI - Description of a clinical pharmacist intervention administered to primary care patients with depression. AB - The objective of this article is to provide a detailed description of interactions between patients with depression and pharmacists. Analysis was conducted on patients from the intervention arm (n=268) of an randomized controlled trial that evaluated the impact of a clinical pharmacist on the outcomes for depressed primary care patients from nine metropolitan Boston practices. The main outcome measure was the amount of intervention time spent with patients, physicians, and other activities. Details of the behavioral intervention and a categorization of the activities are offered. Pharmacists reported 978 encounters with 268 patients in 6 months. Eighty percent of patient encounters occurred by telephone. Initial encounters took 45 min if in person and 13.3 min if by telephone. Subsequent encounters followed a similar pattern. Follow-up visits occurred 2.3 times per patient. Physician contact took considerably less time. In total, the pharmacist intervention took 70.3 min per patient over 6 months; 42.2% of encounters involved an activity related to non antidepressant medication and 85% of encounters involved general support. Other activities (education, advocating antidepressants, and motivating adherence) occurred in at least 50% of encounters. Pharmacists repeated intervention activities in the same category approximately two to three times. Interventions to improve the care of depression in primary care patients must anticipate encountering intense needs for information, personal support, and help negotiating the healthcare system. Research that identifies relationships between the components (active ingredients) of an intervention and the outcomes of care will benefit future intervention strategies and contribute to improved and efficient care. PMID- 15121351 TI - Podiatric problems are associated with worse health status in persons with severe mental illness. AB - The objective of this study was to determine the prevalence of self-reported podiatric impairments and their effect on health status in persons with severe mental illness. A sample of psychiatric outpatients (N=309) underwent interviews assessing medical conditions and health status with the Medical Outcomes Study Short Form-36 (SF-36). Podiatric health was assessed using nine items from the National Health Interview Survey (NHIS). Eighty percent of patients reported at least one podiatric problem. The most common problems were foot pain (48%), nail disorders (35%) and corns/calluses (28%). Prevalence rates were 4-11 times higher than those reported by the general population in the 1990 NHIS. The total number of podiatric problems was inversely related to eight self-reported health status domains and both summary SF-36 scores (all P<==.0001). After controlling for sociodemographic factors, psychiatric illness and medical conditions, the total number of podiatric limitations remained significantly associated with lower patient ratings in four of the eight SF-36 domains and both summary scores. We concluded that persons with severe and persistent mental illness have markedly elevated rates of podiatric problems when compared to the general population group. These problems are associated with worsened self-perceived health status. Addressing podiatric health may be a successful way to improve the overall health of this population. PMID- 15121352 TI - Mood-stabilizer-maintained, remitted bipolar patients: taper and discontinuation of adjunctive antipsychotic medication. AB - The aim of this study was to determine whether bipolar patients who had been stabilized on combined antipsychotic and mood-stabilizer medications and were currently in remission benefited from continuation of the antipsychotic medication. Remitted bipolar patients were randomly assigned to either remain on adjunctive antipsychotic medication or to taper to placebo. Antipsychotic/placebo medication assignment was double-blind. Subjects were outpatients at a university affiliated community mental health center. Fifteen subjects consented and proceeded with eligibility assessments. Five subjects were never randomized. One of these was excluded when the Structured Clinical Interview for DSM-IV interview revealed schizoaffective disorder. The remaining four subjects were not randomized for other reasons. Three randomized subjects never received study medications, or were withdrawn by the investigator within 1 week after beginning study medications. The seven remaining subjects received study medication for more than 1 week. Five subjects were randomized to taper to placebo and two to antipsychotic continuation. Of the five randomized to taper to placebo, three successfully tapered and completed the year of follow-up in continuous remission. One subject became manic 4 months after taper was completed, and one subject became psychotic, in the absence of a mood episode, during taper. Of the two subjects randomized to double-blind antipsychotic continuation, both completed the year of follow-up in continuous remission. When adjunctive antipsychotic medications are discontinued, bipolar patients' clinical symptoms can remain unchanged. Others are, however, at risk for manic relapse. PMID- 15121354 TI - Cortisol and recurrent depressive episodes. PMID- 15121355 TI - A case of social phobia with obsessive-compulsive symptoms improved by paroxetine in combination with risperidone. PMID- 15121353 TI - Suicidal ideation during interferon-alpha2b and ribavirin treatment of patients with chronic hepatitis C. AB - Psychiatric and substance use disorders affect most patients with chronic hepatitis C and are the most common reasons for exclusion from antiviral therapies. Suicidal ideation (SI) is often cited as a reason to exclude patients from interferon-based treatment or to terminate antiviral treatment that is in progress. This study examines SI in hepatitis C patients untreated and treated with interferon-alpha2b, a medication commonly associated with depression. Fifty five subjects with chronic hepatitis C were followed for 24 weeks with three measures of depression, each containing one item assessing SI. A total of 15/55 (27%) subjects reported SI while not on interferon therapy. Of the 42 patients treated with interferon, 18 (43%) endorsed SI at some point during antiviral treatment. However, 17/18 (94%) finished at least a 6-month course of interferon therapy. No subjects attempted suicide. Although SI in some form is common in hepatitis C patients, in most cases it is mild in nature. With adequate support most patients can successfully complete a full course of antiviral treatment. PMID- 15121356 TI - Intense fear of caloric intake related to severe hypoglycemia in anorexia nervosa. PMID- 15121357 TI - Worsening of psychosis or topiramate-induced adverse event? PMID- 15121358 TI - Refractive surgery and protective eyewear in the military. PMID- 15121359 TI - Higher risk of multiple falls among elderly women who lose visual acuity. AB - OBJECTIVE: To determine the association between changes in visual acuity (VA) and frequent falls in older women. DESIGN: Prospective cohort study. PARTICIPANTS: Two thousand two elderly community-residing women participating in the Study of Osteoporotic Fractures with measurements of VA at baseline and a follow-up examination 4 to 6 years later (mean of 5.6 years). METHODS: Binocular VA with habitual correction was measured under standard illumination using Bailey-Lovie charts at baseline and fourth examinations. Change in VA was stratified into 5 categories: no change or VA gain, loss of 1 to 5 letters, loss of 6 to 10 letters, loss of 11 to 15 letters, and loss of >15 letters. A separate analysis considered decline in VA as the loss of >or=10 letters (>or=2 lines) on the Bailey-Lovie acuity measure between baseline and follow-up examinations. MAIN OUTCOME MEASURES: Data on falls were obtained from postcards sent every 4 months after the follow-up examination. Frequent falling was defined as >or=2 falls during a 1-year period after the follow-up examination. RESULTS: Compared with women with stable or improved VA, women with declining acuity had significantly greater odds of experiencing frequent falling during the subsequent year. Odds ratios after adjustment for baseline acuity and other confounders were 2.08 (95% confidence interval [CI]: 1.39-3.12) for loss of 1 to 5 letters, 1.85 (95% CI: 1.16-2.95) for loss of 6 to 10 letters, 2.51 (95% CI: 1.39-4.52) for loss of 11 to 15 letters, and 2.08 (95% CI: 1.01-4.30) for loss of >15 letters. In the analysis of visual decline defined as a loss of >or=10 letters, heightened risk of frequent falling was evident in each of 2 subgroups defined by splitting the sample on baseline VA, with borderline significant evidence of a more pronounced effect in those women with baseline VA of 20/40 or worse (P value for interaction, 0.083). CONCLUSIONS: Loss of vision among elderly women increases the risk of frequent falls. Prevention or correction of visual loss may help reduce the number of future falls. PMID- 15121360 TI - Redundant publications in scientific ophthalmologic journals: the tip of the iceberg? AB - OBJECTIVE: The number of scientific publications is often used to measure scientific achievement. This practice can motivate unethical conduct, such as redundant or duplicate publications, defined as publication of the same scientific contents in more than 1 journal. The aim of this study was to estimate the amount of redundant publications in ophthalmologic journals. DESIGN: Retrospective analysis of published literature. METHODS: We developed an electronic search engine for redundancies to estimate the amount of duplicate publications in scientific journals. When redundancies reached a given degree (matching score), the articles were screened manually based on authors, titles, and abstracts. We applied this method to the 22 433 articles that were published between 1997 and 2000 in 70 ophthalmologic journals indexed by MEDLINE. MAIN OUTCOME MEASURES: The number of duplicate publications with a matching score of 0.6 or more, the number of involved journals, and the number of authors. RESULTS: Redundancies reached a matching score of 0.6 or more in 13 967 pairs of articles. Out of them, a sample of 2210 was reviewed manually. We found 60 redundant articles and estimated that 1.39% of the publications were redundant. Thirty-two journals and an estimate of 1092 authors were involved. In 5% of cases, the scientific conclusions were modified. CONCLUSIONS: Because of the restrictive selection process, the impracticability of detecting all redundant publications, and the estimated amount of duplicates increases with lower matching scores, we regard our estimate to be the tip of the iceberg. Duplicate publications have several negative impacts, but neither peer reviewers nor editors can protect their journal from them completely. Several deterrents for duplicate publications are possible, but as long as publications remain the central requirement for academic advancement, a solution seems unlikely. Nevertheless, it is the responsibility of all those who care about objective research and evidence-based medicine to address this problem-not only in ophthalmology. PMID- 15121361 TI - Videokeratographic anomalies in familial keratoconus. AB - OBJECTIVE: To analyze videokeratography of relatives of established familial keratoconus (FK) patients to detect low-expressivity keratoconus and improve the diagnosis criteria of forme fruste keratoconus. DESIGN: Multicenter case-control study. PARTICIPANTS: Twenty-three families with 55 clinical keratoconus patients, 89 first-degree relatives, 43 other relatives, and a control group of 130 subjects. METHODS: Videokeratography was performed on both eyes of patients after clinical examination, and corneal maps were analyzed. Statistical comparisons were conducted between first-degree and other relatives and a control population. MAIN OUTCOME MEASURES: Qualitative (using a 0.5-diopter [D] increment scale) and quantitative analyses of videokeratographs. RESULTS: Two corneal patterns were overrepresented in the relatives of FK patients: the J and inverted-J form patterns. Results of the quantitative analysis of these suspect patterns showed that the inferior - superior values (reflecting the inferior - superior dioptric asymmetry) were close to 0.8 D and the Srax (relative skewing of the steepest radial axes) was superior to 21 degrees. CONCLUSION: Our study using topography in clearly established genetic keratoconus families allowed us to detect suspect topographical patterns and brings new data to the difficult task of diagnosing forme fruste keratoconus. PMID- 15121362 TI - Central corneal thickness measurements with partial coherence interferometry, ultrasound, and the Orbscan system. AB - OBJECTIVE: To compare the reliability of central corneal thickness measurements (CCT) obtained with partial coherence interferometry (PCI), ultrasound pachymetry, and the Orbscan system. DESIGN: Cross-sectional study. PARTICIPANTS: Twenty healthy subjects with CCT measurements in both eyes. METHODS: The CCT measurements were obtained with PCI, ultrasound pachymetry, and the Orbscan system. In each eye, 2 investigators performed 5 repeated measurements with each pachymetric device. Intraclass correlation coefficients (kappa) were calculated and mean CCT measurements were compared. MAIN OUTCOME MEASURES: The CCT measurements obtained with ultrasound pachymetry, the Orbscan system (Orbtek Inc., Salt Lake City, UT), and PCI. RESULTS: Mean CCT values measured with ultrasound pachymetry were significantly thicker than those measured with PCI (21.5 microm; P<0.001) or the Orbscan system (19.8 microm; P<0.001). The correlation coefficients for the intraobserver variability were 0.999 for PCI measurements, 0.983 for ultrasound pachymetry measurements, and 0.988 for Orbscan system measurements. The correlation coefficients for the interobserver variability were 0.998 for PCI measurements, 0.980 for ultrasound pachymetry measurements, and 0.988 for Orbscan system measurements. There was a slightly better consistency between ultrasound pachymetry and PCI (kappa = 0.96) than between the Orbscan system and PCI (kappa = 0.92) and between ultrasound pachymetry and the Orbscan system (kappa = 0.89). CONCLUSIONS: Partial coherence interferometry was the method with the least intraobserver or interobserver variability. Mean CCT as measured with ultrasound pachymetry was approximately 20 microm thicker than with the Orbscan system and PCI. However, corneal thickness measurements with ultrasound pachymetry and PCI were slightly more consistent than those of the Orbscan system and PCI. This slightly better consistency, however, may be important, especially in corneal refractive surgery. PMID- 15121363 TI - Ultraviolet radiation absorption of intraocular lenses. AB - OBJECTIVES: To record and compare the spectral transmittance curves of intraocular lenses (IOLs) made out of polymethyl methacrylate (PMMA), acrylic, hydrogel, and silicone from different manufacturers; to evaluate their ultraviolet radiation absorption capacities; and to contrast the recorded transmittance curves with that of the natural lens. DESIGN: Experimental study. METHODS: We studied 17 different 21-diopter IOLs. A high-performance spectrophotometer with a diffuse transmittance accessory was employed to measure the transmittance of wavelengths from 200 nm to 800 nm through a 1.5-mm aperture. MAIN OUTCOME MEASURES: Transmittance percentage and 10% transmittance cutoff wavelength. RESULTS: All studied IOLs offered good ultraviolet radiation protection in the ultraviolet C (200-280 nm) and ultraviolet B (280-315 nm) ranges. A number of silicone, PMMA, and acrylic lenses showed different and, at times, only low degrees of absorption in the ultraviolet A (315-400 nm) range. CONCLUSIONS: Intraocular lenses of different compositions have ultraviolet radiation absorption characteristics different from that of the crystalline lens. PMID- 15121364 TI - Anterior capsule relationship of the AcrySof intraocular lens optic and posterior capsule opacification: a prospective randomized clinical trial. AB - OBJECTIVE: To evaluate the relationship of the anterior capsule and the AcrySof MA30BA intraocular lens (IOL) and its impact on the development of central posterior capsule opacification (PCO). DESIGN: Prospective, randomized, controlled trial. PARTICIPANTS: Two hundred two patients with senile cataracts received an AcrySof IOL between July and December 1998 at Iladevi Cataract and IOL Research Center, Ahmedabad, India. INTERVENTION: Patients were randomized prospectively to receive 1 of the 3 possibilities of anterior capsule and IOL optic relationship: group 1, total anterior capsule cover (360 degrees ) of the optic; group 2, no anterior capsule cover (360 degrees ) of the optic; group 3, partial anterior capsule cover (<360 degrees ) of the optic. After surgery, slit lamp video photography was performed every 6 months for 3 years. Analyses of variance and chi-square tests were used to compare treatment groups. MAIN OUTCOME MEASURES: Incidence of PCO in the 3 groups. The posterior capsule was divided into 3 zones: peripheral, central 3 mm, and midperipheral (the space between the peripheral and the central zones). RESULTS: The average follow-up was 35.3 +/- 1.52 months in all the groups. At 3 years, the rate of central PCO was 6.4% in group 1, 7.1% in group 2, and 5.9% in group 3 (P = 0.9). Midperipheral PCO was present in 24.2% in group 1, 16% in group 2, and 20.6% in group 3 (P = 0.9). Peripheral PCO was seen in 100% of patients in all groups. The neodynium:yttrium aluminum-garnet laser (Nd:YAG) posterior capsulotomy rate was 0% in all groups. CONCLUSIONS: There was no significant difference in the incidence of development of central PCO among the 3 groups. No patient experienced central PCO that required Nd:YAG capsulotomy. When using the AcrySof IOL model MA30BA, the relationship of the anterior capsule and the IOL does not seem to be a factor that relates to the development of central PCO. PMID- 15121365 TI - Punctal occlusion in the management of chronic Stevens-Johnson syndrome. AB - PURPOSE: To evaluate the effect of lacrimal punctal occlusion in the management of the ocular surface disease in chronic Stevens-Johnson syndrome. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Thirty-one eyes of 18 patients with chronic Stevens-Johnson syndrome seen at the Department of Ophthalmology of the Tokyo Dental College were studied. INTERVENTION: Patients' lacrimal puncta were occluded by cauterization or with punctal plugs. MAIN OUTCOME MEASURES: The effect of lacrimal punctal occlusion was evaluated by changes in subjective symptoms, best-corrected visual acuity (VA), Schirmer test, tear clearance test, and ocular surface double vital staining before and after punctal occlusion. RESULTS: After lacrimal punctal occlusion, symptomatic improvement was observed in 19 eyes (61.3%). The mean logarithmic VA showed significant improvement from -0.64+/-0.87 to -0.52+/-0.86 (P<0.05). The Schirmer I test results before punctal occlusion yielded a mean score of 7.4+/-8.7 mm, which changed to 10.2+/-8.3 mm after punctal occlusion. The mean preocclusion and postocclusion tear clearance values were 7.5+/-6.6 times and 4.9+/-4.8 times, respectively. The Rose Bengal staining score decreased from 4.7+/-2.8 to 2.7+/ 2.2 points (P<0.05), and the fluorescein staining score decreased from 5.0+/-2.3 to 2.2+/-2.5 points (P<0.05), respectively. CONCLUSIONS: Concurrent improvements in subjective symptoms, vital staining scores, and VA point to the favorable effects of lacrimal punctal occlusion for the ocular surface health in chronic Stevens-Johnson syndrome. PMID- 15121366 TI - Tear function and ocular surface findings in premature and term babies. AB - OBJECTIVE: To describe the ocular surface and tear function findings in premature and term babies. DESIGN: Prospective, case-control study. PARTICIPANTS: Forty eight eyes of 24 premature babies seen at the Department of Ophthalmology of Uludag University School of Medicine, Bursa, Turkey, from March 2002 through September 2002 and 50 eyes of 25 healthy term babies were studied. INTERVENTION: The subjects underwent routine ophthalmic examinations; corneal sensitivity measurements; Schirmer test with anesthesia, with and without nasal stimulation; primary Jones test; fluorescein staining of the ocular surface; and conjunctival impression cytology. MAIN OUTCOME MEASURES: Premature and term babies were compared for corneal sensitivity, lacrimal drainage system patency, tear function and ocular surface staining parameters, goblet cell density, and squamous metaplasia grade. The relation of these parameters to the status of the ocular surface was also investigated. RESULTS: Mean corneal sensitivity scores were 45+/ 5.0 mm and 55+/-4.5 mm in the premature and term babies, respectively (P<0.001). Premature babies had a mean corneal fluorescein staining score of 1.5+/-0.25 points, compared with 0.22+/-0.28 points in the term babies (P<0.001). The mean Schirmer test scores without and with stimulation were 1.5+/-2.5 mm and 4.15+/ 2.5 mm in the premature babies, respectively, compared with 15+/-3.5 mm and 18.75+/-4.5 mm in the term babies. The intragroup and intergroup Schirmer test scores were statistically significant (P<0.001). The primary Jones test was positive in 20.8% of the eyes in the premature babies, whereas it was positive in 84% of eyes in the term babies. The premature babies with positive primary Jones test results all had corneal epithelial defects or severe superficial punctuate keratopathy. Mean conjunctival impression cytology squamous metaplasia scores were 1.86+/-1.2 in the premature babies and 0.86+/-0.47 in the term babies (P<0.001). Mean goblet cell densities were 393+/-484 cells/mm(2) and 739+/-503 cells/mm(2) in the premature and term babies, respectively (P<0.001). CONCLUSION: Decreased corneal sensitivity, reduced tearing, and lacrimal drainage patency are important determinants of ocular surface disease in premature infants. Premature newborns with low Schirmer test scores and a patent lacrimal system may experience corneal and conjunctival epithelial problems and should be carefully checked for the presence of dry eye complications. PMID- 15121367 TI - Persistent fetal vasculature and minimal fetal vascular remnants: a frequent cause of unilateral congenital cataracts. AB - PURPOSE: To determine the significance of persistent fetal vasculature (PFV) and remnants of fetal vessels in the pathogenesis of pediatric unilateral cataracts. STUDY DESIGN: Prospective observational case series. PARTICIPANTS: Thirty-one children with unilateral cataract aged between 2 weeks and 15 years. METHODS: As part of an ongoing prospective clinical trial concerning treatment and etiology of pediatric cataracts, a subgroup of 31 children with unilateral cataracts was defined. The affected eyes received preoperative and intraoperative biomicroscopic examinations to identify characteristic features of PFV and even minimal fetal vascular remnants (MFVRs) at the level of the posterior lens capsule and anterior hyaloid face. In eyes with MFVRs, 3 different severity degrees were assumed, according to different posterior capsule abnormalities: mild, A; moderate, B; and severe, C. All observations were documented on video and analyzed in relation to age (group I, infants between 0 and 1.5 years; group II, preschool children between 1.6 and 5.9 years; group III, schoolchildren between 6 and 16 years). MAIN OUTCOME MEASURES: Frequency and morphology of characteristic features of PFV and MFVRs of the posterior lens capsule/anterior hyaloid face, lens clouding, and microphthalmos. RESULTS: All 31 eyes with unilateral congenital cataracts showed signs of PFV syndrome (100%). Characteristic features of PFV were found in 75% of group I eyes, in 8% of group II eyes, and in 67% of group III eyes. Minimal fetal vascular remnants were found in 25% of group I eyes (severity degree C in all eyes), in 92% of group II eyes (severity degree A in 36.4%, B in 27.2%, and C in 36.4%), and in 33% of group III eyes (severity degree A). Associated microphthalmos was found in all eyes in groups I and III and in 73% of group II, whereas axial lengths were equal in both eyes in 27% of group II children with MFVRs. CONCLUSIONS: Varying degrees of PFV seem to be a frequent cause of unilateral congenital cataracts. Although characteristic features of PFV occurred mainly in infants, eyes of preschool children were usually very mildly affected, showing MFVRs that were detected only by careful observation during surgery. Abnormalities of the central part of the posterior capsule, such as a translucent opacity or a lenticonic area leading to a spontaneous hole during lens aspiration, may be caused by minimal remnants of PFV. PMID- 15121368 TI - Eye care in African Americans with type 1 diabetes: the New Jersey 725. AB - OBJECTIVE: To examine the use of ophthalmologic care by African Americans with type 1 diabetes. DESIGN: Cross-sectional study between 1993 and 1997. PARTICIPANTS: Seven hundred twenty-two African Americans with type 1 diabetes. METHODS: A structured clinical interview was conducted to determine (1) the frequency of annual dilated eye examinations, health insurance, use of an ophthalmologist, and reasons for lack of eye care; and (2) factors associated with having a dilated eye examination by an ophthalmologist during the previous year. All patients also underwent a detailed ocular examination, including masked grading of 7-field stereoscopic fundus photographs. RESULTS: One third of the patients had never been examined by an ophthalmologist. Of those who had eye care, almost half did not have a regular ophthalmologist. During the previous 12 months, only 42% of patients had had a dilated eye examination by an ophthalmologist, despite the fact that approximately three fourths of the patients had been told to do so. The 2 most common reasons given for not seeing an ophthalmologist during the previous year were not having any eye problem (57.6%) and cost (23%). Patients more likely to have had a dilated eye examination by an ophthalmologist during the previous year were married, had higher socioeconomic status, had previously been told about annual dilated eye examinations, and had previously been diagnosed with either a cataract or proliferative diabetic retinopathy. CONCLUSIONS: A large percentage of African Americans with type 1 diabetes do not receive adequate eye care. Screening for diabetic retinopathy and improved access to ophthalmologists are needed to improve eye care in this ethnic group. PMID- 15121369 TI - West African crystalline maculopathy. AB - OBJECTIVE: To report new observations in West African crystalline maculopathy. DESIGN: Retrospective, observational case series. PARTICIPANTS: Three patients drawn from a private retina practice. METHODS: Review of clinical charts and photographic studies. MAIN OUTCOME MEASURES: Distribution of intraretinal crystals and changes after laser photocoagulation, and history of ingesting foods typical in a West African diet but atypical for an American diet. RESULTS: All patients were older than 50 years, had diabetic retinopathy, ate green vegetables not found in American diets, and showed no deleterious effects of the crystals. Kola nut ingestion in 2 patients was remote and sparse, and was unknown in a third patient. The first 2 affected patients originating outside the Ibo tribe of Nigeria are reported. The pattern of retinal crystals can be changed, and the quantity of crystals reduced, by laser photocoagulation of associated diabetic retinopathy. CONCLUSIONS: West African crystalline retinopathy is distinguishable from other causes of crystalline retinopathy. It may reflect a component of the West African diet, seems to have diabetic retinopathy as a promoting factor via breakdown of the blood-retina barrier, and can be modified by laser photocoagulation of diabetic retinopathy. Increased awareness of the condition will allow physicians seeing West African immigrants to make the diagnosis and treat the patients appropriately. PMID- 15121370 TI - Intrapapillary hemorrhage with adjacent peripapillary subretinal hemorrhage. AB - PURPOSE: To describe the clinical features and to present results of new diagnostic methods to help define the cause of the clinical syndrome of intrapapillary hemorrhage with adjacent peripapillary subretinal hemorrhage (IHAPSH). DESIGN: Retrospective review of patients with IHAPSH at presentation seen in 3 centers in Hawaii and Japan. METHODS: We analyzed data including patient demographics, presenting symptoms, initial and final visual acuities, biomicroscopic findings, fundus photographs, and results of available ancillary testing, including fluorescein angiography, B-scan ultrasonography, and optical coherence tomography. RESULTS: There were 10 eyes of 9 patients (7 female and 2 male, 8 Asian and 1 white) aged 14 to 79 years. All patients experienced an acute onset of visual symptoms. Eight eyes had mild to severe myopia (-2.50 diopters [D] to -9.50 D), and 8 eyes had a tilted disc. Hemorrhage within the disc and adjacent subretinal hemorrhage were located nasally in 6 eyes, superiorly in 2 eyes, and temporally in 2 eyes. Vitreous hemorrhage was noted in 6 of 10 eyes. Posterior vitreous evaluation by biomicroscopy (10 eyes), by B-scan ultrasonography (4 eyes), and by optical coherence tomography (2 eyes) revealed no evidence of vitreopapillary traction, except for a follow-up optical coherence tomography in 1 eye showing localized vitreoretinal separation with residual attachment to the optic disc 10 months after presentation. Fluorescein angiography showed mild disc staining in 4 of 8 eyes. Hemorrhage spontaneously resolved within 1 to 7 months, and there were no recurrent hemorrhages with an average follow-up of 13.5 months (range, 2-31 months). Visual acuities maintained or improved to 20/25 or better in 8 eyes. The other 2 eyes had unrelated poor vision. CONCLUSIONS: Intrapapillary hemorrhage with adjacent peripapillary subretinal hemorrhage is more common in myopic eyes and spontaneously resolves without treatment. The unique structural architecture of the elevated nasal edge of the myopic tilted disc and the choroidal blood supply of the prelaminar optic nerve may predispose patients to bleeding from the optic discs, which may be spontaneous or may be precipitated by acute disc edema, Valsalva maneuver, or vitreopapillary traction. PMID- 15121371 TI - Effects of verteporfin therapy on central visual field function. AB - PURPOSE: To evaluate the effect of photodynamic therapy with verteporfin on the maintenance of central visual field function. DESIGN: Randomized controlled clinical trial. PARTICIPANTS: Forty-six consecutive patients with subfoveal choroidal neovascularization (CNV) caused by age-related macular degeneration including a classic component were randomly assigned. Thirty-three participants received standard verteporfin therapy, and 13 received placebo and laser treatment. METHODS: The trial was performed as a single-center, double-masked study. Patients were examined before therapy and continuously in 3-month intervals during 2 years of follow-up. A scanning laser ophthalmoscope (SLO) was used to perform macular microperimetry. Absolute and relative scotomas were documented at each visit, and size was measured in square millimeters. MAIN OUTCOME MEASURES: The change in size of central scotoma in the verteporfin group compared with the placebo group. RESULTS: An absolute scotoma was seen in 88%, and a relative scotoma was seen in 100% of eyes before therapy. Absolute defects were associated with the classic CNV component localized angiographically. In the verteporfin group, the absolute scotoma grew from 2.5 mm(2) at baseline to a final size of 7.3 mm(2) at month 24. In the placebo group, the mean lesion size of the absolute scotoma enlarged from an initial size of 2.7 mm(2) to 31.5 mm(2) after 24 months. The relative scotoma increased from 7.9 mm(2) at baseline to 20.8 mm(2) at month 24 in the verteporfin group, whereas a progression from 8.5 mm(2) initially to 48.3 mm(2) at the final presentation was measured in the placebo group. Statistical analysis showed that both the mean absolute and relative scotoma sizes were significantly smaller in the verteporfin group than the placebo group for all intervals from 6 to 24 months (P<0.001). CONCLUSIONS: Documentation of macular function with SLO perimetry demonstrated a significant benefit of verteporfin therapy for the preservation of the central visual field. Absolute and relative scotoma sizes remained smaller after therapy. This may influence reading ability and visual rehabilitation. PMID- 15121372 TI - Hyperhomocysteinemia and the methylenetetrahydrofolate reductase 677C-T mutation in patients under 50 years of age affected by central retinal vein occlusion. AB - PURPOSE: To investigate the correlation between increased homocysteine plasma levels and the homozygosity for the 677C-T mutation in the methylenetetrahydrofolate reductase (MTHFR) gene in patients aged under 50 years affected by central retinal vein occlusion (CRVO). DESIGN: Prospective, case control study. PARTICIPANTS AND CONTROLS: Participants included 31 consecutive patients under 50 years and diagnosed with CRVO. Two controls per case were selected. The first control group (group I) included 31 individuals matched for age, gender, laboratory tests, and the main risk factors for atherosclerosis. The second control group (group II) consisted of 31 volunteers matched only for age and gender. METHODS: Fasting (>10 hours) blood samples were obtained from patients and controls. Blood samples were obtained from patients within 1 week after the onset of the vaso-occlusive event. Molecular genetic analysis for the 677C-T mutation in the MTHFR gene was performed in patients and controls. A plasma homocysteine reading of >12 micromol/l was considered an increase. MAIN OUTCOME MEASURES: The total homocysteine plasma level (determined by the high performance liquid chromatography method with fluorescence detection) and molecular genetic analysis for the 677C-T mutation in the MTHFR gene in patients and controls. RESULTS: Mean ages were 44.5 years in the group comprising the patients and 44.3 and 44.2 years, respectively, in groups I and II. Mean homocysteine plasma levels were 10.60 micromol/l in patients and 10.39 and 9.34 micromol/l, respectively, in groups I and II. There was no statistically significant difference between mean homocysteine plasma levels in patients and group I controls. In fact, the mean homocysteine plasma level was lower in group II than in patients, and the difference was statistically significant. Homozygosity for the 677C-T mutation in the MTHFR gene was found in 4 patients (12.9%), 5 controls in group I (16.1%), and 4 controls in group II (12.9%). CONCLUSION: The results of the present investigation support the hypothesis that the homocysteine plasma level is not to be considered a primary and independent risk factor for CRVO, but is more likely a marker of atherosclerosis and the consequence of other well-established risk factors. Moreover, the importance of the study design is brought out, because the results we obtained differ on the basis of the considered control group. This feature may in part explain the contradictory results reported in the literature. PMID- 15121373 TI - Patterns of macular edema in patients with uveitis: qualitative and quantitative assessment using optical coherence tomography. AB - PURPOSE: To describe the morphologic characteristics of uveitic macular edema by the use of optical coherence tomography (OCT) and to investigate the correlation between tomographic features and visual acuity (VA). DESIGN: Cross-sectional study. PARTICIPANTS: Seventy consecutive patients with uveitis in a university based practice with a clinical diagnosis of macular edema in at least one eye. Inclusion criteria were: (1) verification of macular edema by OCT, (2) adequate media clarity for fundus visualization, and (3) absence of coexisting ocular disease limiting visual potential. METHODS: Complete ophthalmic examination: best corrected Snellen VA, slit-lamp examination, fundus biomicroscopy, indirect ophthalmoscopy, and OCT. Fluorescein angiography was performed in selected cases. MAIN OUTCOME MEASURES: Best-corrected Snellen VA and tomographic features of the macula, including macular thickness measurement and correlation of macular thickness with VA. RESULTS: Eighty-four eyes of 60 patients met the inclusion criteria. The mean VA (Snellen test) was 20/36. There were 3 patterns of macular edema: diffuse macular edema (DME), cystoid macular edema (CME), and serous retinal detachment (RD). Serous retinal detachment was detected in 17 eyes (20.2%). Patients were classified into the following groups: DME (46 eyes, 54.8%), CME (21 eyes, 25%), DME and RD (5 eyes, 5.9%), CME and RD (12 eyes, 14.3%). Epiretinal membrane was detected by OCT in 34 eyes (40.5%). Eight eyes (9.5%) demonstrated vitreomacular traction. The mean retinal thickness at the central fovea was 333+/-171 microm (mean +/- standard deviation). Macular edema was located mainly in the outer retinal layers. Eyes with CME had significantly greater retinal thickness measurements than eyes with DME (P<0.001). Multivariate analysis revealed that VA was negatively correlated with increased macular thickness, presence of CME, and RD (P<0.05). CONCLUSIONS: Optical coherence tomography demonstrated 3 patterns of macular edema in patients with uveitis: DME, CME, and RD. Epiretinal membrane coexisted in a significant percentage of patients. In patients with uveitis with clear media, the morphologic features of macular edema and macular thickness correlated with VA. PMID- 15121374 TI - Birdshot chorioretinopathy: long-term manifestations and visual prognosis. AB - PURPOSE: To ascertain the clinical features and long-term visual prognosis of birdshot chorioretinopathy (BCR), and to identify patients at risk of visual loss. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Fifty-five consecutive patients with HLA-A29-positive BCR who were identified in ophthalmology departments of the University Medical Center of Utrecht and The Eye Hospital Rotterdam, of whom 37 were observed for at least 5 years. INTERVENTION: A review of the medical and photographic and/or angiographic records of 55 patients with HLA-A29-positive BCR. MAIN OUTCOME MEASURES: Numerous variables were compared, including age and gender distribution, onset and course of BCR, ocular manifestations, therapeutic strategies and their outcomes, complications, systemic diseases, visual acuity (VA), and features associated with poor visual outcome. RESULTS: Loss of VA was gradual; the number of affected eyes with VA less than 20/200 increased from 9 of 108 (8%) at onset to 22 of 73 (30%) at 5 years and 19 of 49 (39%) at 10 years of follow-up. The cause of compromised VA was predominantly macular edema and macular atrophy (42 of 55 [76% of cases]). We found strong associations between the VA at onset and visual outcome after 5 and 10 years (P = 0.005 and P = 0.006, respectively). Mean VA at the 5-year follow-up was significantly lower if macular leakage was observed on angiography (P<0.001). No differences in annual loss of VA were observed between patients treated by standard therapeutic modalities and untreated patients. CONCLUSION: The visual prognosis of BCR in a spectrum of uveitis is poor, and the recommended therapeutic regimens have had no effect on long-term visual prognosis. New treatment strategies are needed for this blinding disorder. PMID- 15121375 TI - Pulse IV cyclophosphamide in ocular inflammatory disease: efficacy and short-term safety. AB - PURPOSE: To assess the efficacy and short-term safety of appropriately monitored pulse IV cyclophosphamide therapy in the treatment of patients with severe or treatment-resistant autoimmune ocular inflammatory disease. DESIGN: Retrospective noncomparative interventional case series. PARTICIPANTS: Thirty-eight patients with severe or recalcitrant ocular inflammation of diverse etiologies. METHODS: Charts of patients seen on the Ocular Immunology & Uveitis Service at the Massachusetts Eye & Ear Infirmary were reviewed. Thirty-eight consecutive patients treated with pulse IV cyclophosphamide between January 1995 and March 2002 were analyzed. MAIN OUTCOME MEASURES: The control of inflammation, steroid sparing effect, visual acuity, and adverse reactions. RESULTS: A positive response to treatment occurred in 68% of patients during the study period, with 55% achieving complete quiescence. A steroid-sparing effect was achieved in all patients previously on systemic steroid, allowing successful discontinuation of the drug in 41%. Visual acuity was maintained in 66% and improved in 21% of involved eyes. The most common side effects observed were fatigue (63%), nausea (32%), and headache (22%). None required a permanent discontinuation of therapy. CONCLUSIONS: Pulse IV cyclophosphamide is an effective therapeutic modality in patients with severe or treatment-resistant ocular inflammatory disease. PMID- 15121377 TI - Conservation of eyes with choroidal melanoma by a multimodality approach to treatment: an audit of 1632 patients. AB - OBJECTIVE: To report on conservation of eyes with choroidal melanoma with a multimodality approach to treatment. DESIGN: Prospective, noncomparative, interventional case series. PARTICIPANTS: All 1632 patients with choroidal melanoma treated at a single center between 1993 and 2002. INTERVENTION: Primary enucleation (35%), brachytherapy (31.3%), proton beam radiotherapy (16.7%), transscleral local resection (11.0%), endoresection (3.7%), transpupillary thermotherapy (2.5%), and photocoagulation (0.1%). MAIN OUTCOME MEASURES: Primary and secondary enucleation. RESULTS: Logistic regression showed the main predictive factors for primary enucleation to be: age more than 60 years (odds ratio [OR], 2.4; 95% confidence interval [CI], 1.8%-3.2%); reduced visual acuity (OR, 2.5; 95% CI, 1.9%-3.2%); posterior extension close to or involving the optic disc and fovea (OR, 1.7; 95% CI, 1.2%-2.4%); circumferential spread around the ciliary body, iris, or angle (OR, 3.1; 95% CI, 1.8%-5.5%); basal tumor diameter (OR, 3.5; 95% CI, 2.4%-5.0%); and tumor height (OR, 6.3; 95% CI, 4.5%-8.9%). After conservative treatment, the actuarial rate of secondary enucleation was 11.1% at 5 years (95% CI, 8.6%-13.6%). Cox multivariate analysis indicated the factors independently predicting secondary enucleation as: nasal/midline tumor location (risk ratio [RR], 2.6; 95% CI, 1.6%-4.4%); disc involvement (RR, 2.2; 95% CI, 1.2%-4.1%); tumor diameter (RR, 1.2; 95% CI, 1.0%-1.5%); and tumor thickness (RR, 1.8; 95% CI, 1.5%-2.1%). CONCLUSIONS: With a multimodality approach to treatment, 65% of patients underwent conservative treatment, and of them, 89% retained their eye at 5 years, with success depending on tumor diameter, tumor thickness, disc involvement, and coronal location. PMID- 15121376 TI - Ten-year follow-up of fellow eyes of patients enrolled in Collaborative Ocular Melanoma Study randomized trials: COMS report no. 22. AB - PURPOSE: To report findings observed in fellow eyes during prospective follow-up of patients with unilateral choroidal melanoma after treatment with standard enucleation or 1 of 2 radiotherapy methods, either iodine 125 (I(125)) brachytherapy or pre-enucleation external radiation, in order to document long term outcomes and to identify any adverse effect of radiotherapy on the contralateral eye. DESIGN: Two multicenter randomized trials conducted by the Collaborative Ocular Melanoma Study (COMS) Group. PARTICIPANTS: Eligible patients assigned randomly to standard enucleation or to the radiotherapy protocol adopted for tumors of the specified size and location and treated as assigned: 994 patients of 1003 enrolled in the COMS trial of pre-enucleation radiation and 1296 patients of 1317 enrolled in the COMS trial of I(125) brachytherapy. OUTCOMES: Changes in best-corrected visual acuity (VA), intraocular pressure, and other findings in fellow eyes from baseline to examinations conducted at 6 and 12 months after enrollment and annually thereafter. RESULTS: Five years after enrollment, 1307 of 2290 fellow eyes were examined; 358 fellow eyes were examined 10 years after enrollment. Mean change in VA of fellow eyes from baseline to each examination was one letter (0.2 lines) or less. Cumulative 5-year incidence rates of cataract surgery and visually significant cataract in initially phakic eyes with good VA and no lenticular opacity were 8% in both trials; 10-year rates were 18% in the trial of pre-enucleation and 15% in the trial of I(125) brachytherapy. Intraocular pressures changed by less than 1 mmHg from baseline to each examination. Apart from lower rates of incident cataracts among fellow eyes of patients treated with pre-enucleation radiation, findings within each trial were similar in the 2 treatment arms. CONCLUSIONS: Almost all surviving patients retained good VA in fellow eyes throughout 5 years of follow-up after treatment for choroidal melanoma. These findings persisted through 10 years of follow-up among patients eligible for examinations beyond 5 years. There was no evidence that fellow eyes of patients whose affected eye was treated with pre-enucleation radiation or with I(125) brachytherapy were at greater risk of loss of VA or new ophthalmic diagnoses than eyes of patients treated with enucleation alone. PMID- 15121378 TI - Transpupillary thermotherapy as initial treatment for small intraocular retinoblastoma: technique and predictors of success. AB - OBJECTIVE: To determine which small retinoblastoma tumors can be treated with transpupillary thermotherapy (TTT) alone and to clarify the minimum amount of treatment required to prevent recurrence. DESIGN: Noncomparative interventional case series. PARTICIPANTS: Ninety-one tumors in 22 eyes of 24 patients were treated with TTT as the primary treatment modality at a single institution from 1995 until 2002. METHODS: Transpupillary thermotherapy was applied only when a tumor first appeared or if growth subsequently occurred. Treatment was performed while patients were under general anesthesia with the Iris diode laser (810 nm) on continuous mode with a 1.2-mm spot size. MAIN OUTCOME MEASURES: Local tumor recurrence, failure of TTT requiring the use of salvage therapies. RESULTS: Mean age at diagnosis was 3 months (range, 0-19 months), and mean initial tumor base was 0.67 disc diameters (DD) (range, 0.1-1.5). Eighty-four tumors (92%) were cured with TTT alone. Seven tumors (8%) required between 1 and 5 salvage treatments: 4 tumors received cryotherapy, 3 tumors received systemic chemotherapy, 2 tumors received external beam radiation, and 1 tumor received periocular chemotherapy. All 7 tumors requiring salvage treatment were cured without enucleation. Twenty-one eyes were preserved (95%), with 1 eye enucleated secondary to growth of other tumors. The mean number of treatment sessions required for cure was 1.7 (median, 1 session, range 1-6), with 64% of the tumors requiring only 1 session. The mean power was 420 mW, the mean total duration for all sessions was 320 seconds, and the mean total energy for all treatments was 139.6 J. Univariate analysis revealed that predictors of tumor recurrence were male gender, increasing age at diagnosis, posterior and inferior tumor location, increasing initial tumor base diameter, and increasing total energy. Multivariate analysis indicated that the predictive combination of variables included male gender, increasing age at diagnosis, and increasing total energy. The predictors of need for salvage treatment on univariate analysis were male gender, inferior tumor location, increasing initial tumor base diameter, and increasing total energy. Multivariate analysis demonstrated that the most important combination of variables was male gender and increasing total energy. CONCLUSIONS: Retinoblastoma tumors <1.5 DD in base diameter can be successfully treated with TTT alone. Treatment can be implemented only when recurrences occur and can be used at low power settings for a short duration. PMID- 15121379 TI - Chorioretinal scar growth after 810-nanometer laser treatment for retinoblastoma. AB - PURPOSE: To determine the extent of expansion of laser scars after single treatment with transpupillary thermotherapy (TTT) for retinoblastoma. DESIGN: We retrospectively reviewed medical records and RetCam (Massie Industries, Dublin, CA) digital fundus photographs of patients with retinoblastoma who received TTT by use of an 810-nm diode laser with large-spot indirect ophthalmoscope. Digital images were measured by use of RetCam software beginning immediately after treatment. Lesions were measured in both linear size and area by 2 observers. In addition, optic disc diameters were also measured for each image. PARTICIPANTS: We identified 9 patients with hereditary retinoblastoma who were treated from 1997 to 2000. MAIN OUTCOME MEASURES: Linear and area expansion of laser scars as a function of time. RESULTS: Fourteen tumors in 10 eyes were treated successfully with 1 session of TTT. Follow-up for all eyes was at least 16 months. Scars of 12 of the 14 lesions increased beyond their original borders (mean starting linear diameter, 2.02 mm; range 1.46-2.59 mm; mean increase, 0.72 mm [36%]) over time, most within 6 months to 1 year after treatment. CONCLUSIONS: Scars from TTT for retinoblastoma may increase in size after treatment. This expansion must be considered when applying TTT to tumors near vital macular structures, such as the fovea and optic nerve. PMID- 15121380 TI - Survey of 1264 patients with orbital tumors and simulating lesions: The 2002 Montgomery Lecture, part 1. AB - OBJECTIVE: To determine the incidence of specific orbital tumors based on patients referred to an ocular oncology center. DESIGN: Retrospective, observational case series. PARTICIPANTS: A total of 1264 consecutive patients referred to an ocular oncology service because of space-occupying orbital lesions. METHODS: A retrospective chart review was carried out for 1264 consecutive patients referred for a suspected orbital mass over a 30-year period. The lesions were grouped into general categories, as shown in "Results." The specific diagnosis in each case was based on clinical findings, computed tomography scan results, magnetic resonance imaging results, and histopathologic analysis results, when available. The number and percentage of benign and malignant tumors per age group also was determined. MAIN OUTCOME MEASURES: The incidence of orbital tumors and pseudotumors. RESULTS: Among 1264 consecutive patients, the number and percentage of lesions in each general category were as follows: cystic, 70 cases (6%); vasculogenic, 213 cases (17%); peripheral nerve lesions, 23 (2%); optic nerve and meningeal tumors, 105 (8%); fibrocytic lesions, 13 (1%); osseous and fibro-osseous tumors, 21 (2%); cartilaginous lesions, 1 (<1%); lipocytic and myxoid lesions, 64 (5%); myogenic tumors, 36 (3%); lacrimal gland lesions, 114 (9%); primary melanocytic lesions, 11 (<1%); metastatic tumors, 91 (7%); lymphoma and leukemia lesions, 130 (10%); secondary orbital tumors, 142 (11%); histiocytic lesions, 17 (1%); thyroid-related orbitopathy, 67 cases (5%); other inflammatory lesions, 133 cases (11%); and miscellaneous other lesions, 13 (1%). The most common diagnoses were: lymphoid tumor (139 cases;11%), idiopathic orbital inflammation (135 cases; 11%), cavernous hemangioma (77 cases; 6%), lymphangioma (54 cases; 4%), meningioma (53 cases; 4%), optic nerve glioma (48 cases; 4%), metastatic breast cancer (44 cases;4%), orbital extension of uveal melanoma (41 cases; 3%), capillary hemangioma (36 cases;3%), rhabdomyosarcoma (35 cases; 3%), dermolipoma (31 cases; 3%), herniated orbital fat (30 cases; 2%), dermoid cyst (26 cases; 2%), varix (26 cases; 2%), dacryops (19 cases; 2%), and other less common lesions. Of the 1264 lesions, 810 (64%) were benign and 454 (36%) were malignant. The percentage of malignant lesions was 20% in children (age range, 0-18 years), 27% in young adults and middle-aged patients (age range, 19-59 years), and 58% in older patents (age range, 60-92 years). Rhabdomyosarcoma was the most common malignancy in children, representing 3% of all orbital masses, and lymphoma was the most common malignancy in older patients, representing 10% of cases. CONCLUSIONS: A variety of tumors and pseudotumors can involve the orbit. In this series of 1264 lesions, 64% were benign and 36% were malignant. The percentage of malignant tumors increased with age, with malignancies being common in older patients because of the higher incidence of lymphoma and metastasis in the elderly. PMID- 15121381 TI - Rapid intraoperative diagnosis of tumors of the eye and orbit by squash and imprint cytology. AB - PURPOSE: To evaluate the diagnostic accuracy of intraoperative squash and imprint cytologic techniques in ocular and orbital lesions. DESIGN: Observational case series. METHODS: Forty-five consecutive ocular and orbital biopsy specimens subjected to squash and imprint cytologic techniques for rapid intraoperative diagnosis were included in this study. The indications for which the surgeon requested intraoperative diagnosis included: (1) infiltrative lesions where a preoperative tissue diagnosis was not available, (2) discrepancy between preoperative and intraoperative clinical diagnosis, and (3) unusual clinical presentations with an uncertain diagnosis. Fresh unfixed tissue was used for making squash preparations and imprint smears. Alcohol-fixed smears were stained with hematoxylin and eosin, whereas air-dried smears were stained with the Giemsa stain. The residual tissue was subjected to routine processing for histopathologic examination. MAIN OUTCOME MEASURE: Correlation of intraoperative cytologic diagnosis with the final histologic diagnosis was the primary outcome measure. The correlation was considered to have complete concordance if the diagnosis matched with respect to nature of the lesion (benign vs. malignant) as well as with the further classification of the benign or the malignant lesion. Partial concordance was considered when there was agreement with respect to the nature of the lesion, but there was a mismatch in the further classification of the benign or the malignant lesion. RESULTS: Biopsies were obtained from 45 patients whose ages ranged from 1 to 79 years (median, 21 years). The sites of biopsy included eyelids (n = 4), conjunctiva (n = 6), intraocular tissue (n = 3), and orbit (n = 32). Adequate cellularity was obtained in all cases by squash and imprint techniques. A cytologic diagnosis could be made within 5 to 8 minutes after receiving the sample. Of 44 cases in which a cytologic diagnosis could be made, 21 (48%) were malignant and 23 (52%) were benign. Cytologic analysis was inconclusive in 1 case. The cytologic-histologic correlation showed complete concordance in 41 of 45 cases (91%) and partial concordance in 3 cases. Sensitivity and specificity of the squash and imprint technique in detecting malignant lesions was 100%. CONCLUSIONS: Assuming the availability of a skilled cytologist, ophthalmic pathologist, or both, squash and imprint preparations are useful for rapid and reliable intraoperative diagnosis of ocular and orbital lesions. PMID- 15121382 TI - Congenital mesenchymal chondrosarcoma of the orbit: case report and review of the literature. AB - PURPOSE: To report the first case of congenital extraskeletal mesenchymal chondrosarcoma arising in the orbit. DESIGN: Interventional case report and review of the literature. METHODS: Ophthalmologic examination and computed tomography scan of the orbit were performed. Histologic and histochemical examination and immunohistochemistry of the biopsy specimen were evaluated. MAIN OUTCOME MEASURES: Ocular and systemic disease control. RESULTS: A 5-month-old girl was seen with a papillomatous lesion in the right lower tarsal conjunctiva present from birth. Histologic examination demonstrated an admixture of undifferentiated mesenchymal cells and islands of mature hyaline cartilage, and immunohistochemistry studies revealing positivity for vimentin and S-100 were consistent with the diagnosis of mesenchymal chondrosarcoma. The patient was treated with combined chemotherapy (ifosfamide, epirubicin, and cisplatin), radiotherapy, and surgery (exenteration). She is alive with no evidence of disease after 4 years of follow-up. Facial asymmetry and dental malformations have developed as late effects. Further reconstructive surgery is planned for the malformations. CONCLUSIONS: Extraskeletal mesenchymal chondrosarcoma in the orbit is extremely rare. This study reports the first case of congenital extraskeletal mesenchymal chondrosarcoma arising in the orbit (also the youngest patient) both in skeletal and extraskeletal sites. Multimodality treatment (surgery, chemotherapy, and radiotherapy) may lead to long-term survival. PMID- 15121383 TI - Orbital myositis associated with Borrelia burgdorferi (Lyme disease) infection. AB - OBJECTIVE: To report on the clinical findings in a patient with isolated left inferior rectus myositis associated with serologically confirmed Borrelia burgdorferi infection. DESIGN: Interventional case report. TESTING: Comprehensive clinical, laboratory, and imaging evaluation. RESULTS: Contrast-enhanced computed tomography showed a swollen inferior rectus muscle with infraorbital soft tissue swelling in a patient with diplopia and prior symptoms consistent with manifestations of Lyme disease. Positive serum and cerebrospinal fluid antibodies to B. burgdorferi by enzyme-linked immunoassay were confirmed by Western blot, and the cerebrospinal fluid/serum antibody ratio was elevated. No alternative cause for orbital myositis was found, and treatment with antibiotics resulted in a complete recovery. CONCLUSIONS: Orbital myositis should be added to the expanding list of ophthalmic manifestations of Lyme disease. Correct diagnosis and appropriate antibiotic therapy may reduce the likelihood of further neurologic or ophthalmologic sequelae. PMID- 15121384 TI - Eyelid position measurement in Graves' ophthalmopathy: reliability of a photographic technique and comparison with a clinical technique. AB - PURPOSE: To evaluate intraobserver reliability and interobserver reliability of a computer-based digital image measurement of eyelid position in Graves' ophthalmopathy and to compare digital image measurement with clinical measurement. DESIGN: Cross-sectional study. PARTICIPANTS: Eighty-four eyes of 42 patients with mild to moderate bilateral Graves' ophthalmopathy. METHODS: Digital images were created from 35-mm color slides of both eyes of participants and projected onto a 15-inch flat-screen computer monitor. Three observers (2 oculoplastic surgeons and 1 ophthalmology resident) independently recorded eyelid fissure height, margin-reflex distance, and inferior scleral show for each eye. MAIN OUTCOME MEASURES: Intraobserver reliability and interobserver reliability of eyelid parameter measurements, as described by the intraclass correlation coefficient (ICC) and Bland-Altman plots. Agreement between digital image measurements of the investigators and clinical measurements taken on the same day as the photographs also was assessed. RESULTS: Excellent intraobserver agreement was found for the measurement of all eyelid parameters for all 3 investigators (ICC range, 0.93-0.99). Interobserver agreement for all eyelid parameters was also excellent for all investigators (ICC, 0.86-0.97). Agreement between the photographic and clinical measurements for eyelid parameters was fair to moderate (ICC range, 0.38-0.62). CONCLUSION: Measurement of several eyelid parameters in Graves' ophthalmopathy patients from computer-based digital images is reliable. Associations between photographic and clinical measurements for all parameters are weaker. Relative to clinical measurements, the photographic technique offers the advantages of potential for masking and ease of transmission that might be useful in clinical trials. PMID- 15121385 TI - Classification and surgical management of patients with familial and sporadic forms of congenital fibrosis of the extraocular muscles. AB - PURPOSE: To outline the clinical features and surgical treatment of patients with familial and sporadic (simplex) forms of congenital fibrosis of extraocular muscles (CFEOM) from 2 countries, and to classify them according to phenotype and mode of inheritance. DESIGN: Observational and experimental study. METHODS: Twenty-eight affected individuals from 20 families with familial or sporadic CFEOM underwent assessment of ocular motility, visual acuity, slit-lamp biomicroscopy, tonometry, and ophthalmoscopy. Seventeen patients had a variety of eye muscle procedures and ptosis repair. RESULTS: There were 1 Iranian family with autosomal dominant CFEOM, 4 Iranian families with autosomal recessive disease, and 15 simplex cases with various CFEOM phenotypes. Two simplex patients had unilateral disease. All other cases were bilateral. Inferior rectus recession improved hypotropia and Bell's phenomenon in the patients with infraducted eyes and chin elevation. Horizontal muscle recession, sometimes combined with opposite muscle resection, corrected horizontal strabismus satisfactorily in most cases. Ptosis was repaired by frontalis sling and/or levator resection. CONCLUSION: Definite recessive CFEOM was present only in Iranian patients. American patients had the classic phenotype of dominant CFEOM, which was also observed in some of the Iranian patients. The surgical management of patients with CFEOM is challenging. Correction of vertical and horizontal strabismus was addressed using large muscle recessions. Ptosis repair should aim at placing the lid level 1 to 2 mm above the pupil in the primary position to avoid exposure keratopathy. PMID- 15121386 TI - Ptosis from localized A-lambda-amyloid deposits in the levator palpebrae muscle. AB - PURPOSE: To describe a patient with unilateral ptosis caused by localized amyloid deposits of immunoglobulin lambda-light chains (A-lambda-amyloid) in the levator palpebrae muscle. STUDY DESIGN: Case report and literature review. PARTICIPANT: A 32-year-old otherwise healthy female with right-sided unilateral ptosis. METHODS: The patient presented with right-sided unilateral ptosis without other ocular abnormalities. Palpebral fissures measured 5 mm on the right side and 8 mm on the left. Systemic evaluation, including neurologic and neuroradiologic examinations, was normal. RESULTS: Several years after initial presentation, conjunctival amyloid deposits appeared. Ptosis surgery with resection of the levator muscle was performed, revealing massive deposits of A-lambda-amyloid in the excised tissue. CONCLUSIONS: Our patient had unilateral ptosis due to localized A-lambda amyloid deposits in the levator muscle. No other amyloid deposits were initially found. Visible conjunctival deposits developed only years later. Ocular amyloidosis with isolated initial involvement of the levator muscle is a rare condition. Localized monoclonal gammopathy may have been the cause of this unique presentation. PMID- 15121387 TI - Localization of extraocular muscles during secondary orbital implantation surgery: the tunnel technique: experience in 100 patients. AB - PURPOSE: To describe a technique for localizing extraocular muscles during secondary orbital implantation and to report the author's experience in 100 consecutive cases. DESIGN: Retrospective, noncomparative, consecutive case series. PARTICIPANTS AND METHODS: A retrospective review was performed of the charts from one surgeon's practice of 100 consecutive patients who received a secondary porous orbital implant for anophthalmic socket rehabilitation. The following data were recorded: age, gender, reason for seeking secondary implantation, type of implant used, size and style of implant used, number of muscles localized, follow-up duration, and problems encountered and their treatment. Preoperative and postoperative photographs, implant motility, prosthetic motility, and overall impressions (by patient and physician) of the surgical outcome were analyzed. RESULTS: The 100 patients (66 males and 34 females, with a mean age of 38.9 years) were followed up for an average of 16.9 months (range, 4-60 months). All 4 rectus muscles were identified in 93 patients, 3 recti were identified in 6 patients, and 2 recti were identified in 1 patient. In 2 patients, the levator muscle mistakenly was thought to be the superior rectus muscle and was connected to the implant. Reconstructive surgical procedures were required in both cases. At final follow-up, 98% of patients believed that they had benefited from surgery, with an improved prosthetic appearance, improved symmetry between the 2 eyes, improved comfort, better prosthetic motility, or a combination thereof. Compared with the results of the preoperative examination, implant and prosthetic motility was improved in 92% to 98% of patients. CONCLUSIONS: The tunnel technique allows successful localization of extraocular muscles during secondary orbital implant surgery in a high percentage of patients. Surgical outcomes were excellent, with few complications and high patient satisfaction. PMID- 15121388 TI - Single-field fundus photography for diabetic retinopathy screening: a report by the American Academy of Ophthalmology. AB - OBJECTIVE: To evaluate whether single-field fundus photography can be used as a screening tool to identify diabetic retinopathy for referral for further ophthalmic care. METHODS: A MEDLINE search of the peer-reviewed literature was conducted in June 2001 for the years 1968 to 2001 and updated in September 2003, yielding 145 articles. The search was limited to articles published in English. The Cochrane Library of clinical trials was also investigated. The authors reviewed the abstracts of these articles and selected 63 of possible clinical relevance for review by the panel. Of these 63 articles, the panel selected 32 for the panel methodologist to review and rate according to the strength of evidence. RESULTS: Three of the 32 articles reviewed were classified as level I evidence, and 4 were classified as level II evidence. Evidence from level I studies demonstrates that as a tool to detect vision-threatening retinopathy, single-field fundus photography interpreted by trained readers has sensitivity ranging from 61% to 90% and specificity ranging from 85% to 97% when compared with the gold standard reference of stereophotographs of 7 standard fields. When compared with dilated ophthalmoscopy by an ophthalmologist, single-field fundus photography has sensitivity ranging from 38% to 100% and specificity ranging from 75% to 100%. CONCLUSIONS: Single-field fundus photography is not a substitute for a comprehensive ophthalmic examination, but there is level I evidence that it can serve as a screening tool for diabetic retinopathy to identify patients with retinopathy for referral for ophthalmic evaluation and management. The advantages of single-field fundus photography interpreted by trained readers are ease of use (only one photograph is required), convenience, and ability to detect retinopathy. Further studies will be required to assess the implementation of single-field photography-based programs to confirm the clinical and cost effectiveness of these techniques in improving population visual outcomes. Future research also should include establishing standardized protocols and satisfactory performance standards for diabetic retinopathy screening programs. PMID- 15121389 TI - Patching regimens. PMID- 15121390 TI - Patching regimens. PMID- 15121392 TI - Capsular tension ring removal. PMID- 15121394 TI - Viscocanalostomy versus trabeculectomy. PMID- 15121396 TI - Extraorbital versus extraconal. PMID- 15121397 TI - Europe's legacy to its children--a healthier environment? PMID- 15121398 TI - Gender-based violence and HIV/AIDS: recognising links and acting on evidence. PMID- 15121399 TI - Immunomodulatory functions of mesenchymal stem cells. PMID- 15121400 TI - Fish oil--an appetising alternative to anti-arrhythmic drugs? PMID- 15121401 TI - Registering clinical trials: an essential role for WHO. PMID- 15121402 TI - Gender-based violence, relationship power, and risk of HIV infection in women attending antenatal clinics in South Africa. AB - BACKGROUND: Gender-based violence and gender inequality are increasingly cited as important determinants of women's HIV risk; yet empirical research on possible connections remains limited. No study on women has yet assessed gender-based violence as a risk factor for HIV after adjustment for women's own high-risk behaviours, although these are known to be associated with experience of violence. METHODS: We did a cross-sectional study of 1366 women presenting for antenatal care at four health centres in Soweto, South Africa, who accepted routine antenatal HIV testing. Private face-to-face interviews were done in local languages and included assessement of sociodemographic characteristics, experience of gender-based violence, the South African adaptation of the Sexual Relationship Power Scale (SRPS), and risk behaviours including multiple, concurrent, and casual male partners, and transactional sex. FINDINGS: After adjustment for age and current relationship status and women's risk behaviour, intimate partner violence (odds ratio 1.48, 95% CI 1.15-1.89) and high levels of male control in a woman's current relationship as measured by the SRPS (1.52, 1.13-2.04) were associated with HIV seropositivity. Child sexual assault, forced first intercourse, and adult sexual assault by non-partners were not associated with HIV serostatus. INTERPRETATION: Women with violent or controlling male partners are at increased risk of HIV infection. We postulate that abusive men are more likely to have HIV and impose risky sexual practices on partners. Research on connections between social constructions of masculinity, intimate partner violence, male dominance in relationships, and HIV risk behaviours in men, as well as effective interventions, are urgently needed. PMID- 15121403 TI - Red ears as red herring. PMID- 15121404 TI - Asbestos exposure as a risk factor for retroperitoneal fibrosis. AB - BACKGROUND: Retroperitoneal fibrosis (RPF) is an uncommon disease with unknown causation in most cases. The pathognomonic finding is a fibrous mass covering the abdominal aorta and the ureters. Our aim was to clarify the possible role of asbestos exposure in the development of RPF. The hypothesis was based on the ability of asbestos to cause fibrosis in pulmonary and pleural tissue. METHODS: We undertook a case-control study of 43 patients with the disease (86% of eligible cases) treated in three university hospital districts of Finland in 1990 2001. For every patient, five population-based controls were selected, matched by age, sex, and central hospital district. We assessed asbestos exposure and medical history using a postal questionnaire and a personal interview. Of the 215 eligible controls, 179 (83%) participated in the study. FINDINGS: The age standardised incidence of RPF was 0.10 (95% CI 0.07-0.14) per 100?000 person years. The disease was strongly associated with asbestos exposure. The odds ratio (OR) was 5.54 (1.64-18.65) for less than 10 fibre-years of asbestos exposure and 8.84 (2.03-38.50) for 10 or more fibre-years, the attributable fraction being 82% and 89%, respectively. Other risk factors were previous use of ergot derivates (OR 9.92 [1.63-60.26]), abdominal aortic aneurysm (OR 6.73 [0.81-56.08]), and smoking for more than 20 pack-years (OR 4.73 [1.28-17.41]). INTERPRETATION: Our results show that occupational asbestos exposure is an important causal factor for RPF. For patients with work-related asbestos exposure, RPF should be considered an occupational disease. PMID- 15121405 TI - Skeletal effects and functional outcome with olpadronate in children with osteogenesis imperfecta: a 2-year randomised placebo-controlled study. AB - BACKGROUND: Non-randomised studies have suggested beneficial effects of bisphosphonates in osteogenesis imperfecta. We assessed the effects of oral olpadronate in children with this disorder in a randomised double-blind placebo controlled trial. METHODS: 34 children recruited from the Dutch national centre for osteogenesis imperfecta were randomly assigned olpadronate (10 mg/m2 daily; n=16) or placebo (n=18) for 2 years. All children also received calcium and vitamin D supplements. Primary endpoints were incident fractures of long bones and changes in bone mineral content (BMC), bone mineral density (BMD), and functional outcome. Anthropometry, vertebral height, and urinary markers of bone resorption were also studied. Analyses were by intention to treat. FINDINGS: Fracture follow-up was complete for all the children, including two who withdrew from the study (one from each group). Olpadronate treatment was associated with a 31% reduction in relative risk of fracture of long bones (hazard ratio 0.69 [95% CI 0.52-0.91], p=0.01). The olpadronate group showed significantly greater increases than the placebo group in spinal BMC (difference between groups 2.24 g/year [0.20-4.29], p=0.03) and spinal BMD (difference between groups 0.054 g/cm2 per year [0.012-0.096], p=0.01). There were no detectable effects on functional outcome, anthropometrics, or vertebral height and no differences between the groups in changes in urinary markers of bone resorption. INTERPRETATION: Oral treatment with olpadronate at a daily dose of 10 mg/m2 results in a reduction of fracture risk of long bones in children with osteogenesis imperfecta. However, the issue of whether bisphosphonates will alter the natural course of osteogenesis imperfecta remains unresolved, and further studies are needed. PMID- 15121406 TI - Bone marrow transdifferentiation in brain after transplantation: a retrospective study. AB - BACKGROUND: End-organ repair by adult haemopoietic stem cells is under great scrutiny with investigators challenging the notion of these cells' plasticity. Some investigations of animals and short-term human bone marrow transplants suggest that bone marrow can repair brain. We looked for evidence of clinically relevant marrow-derived restorative neurogenesis: long-term, multilineage, neural engraftment that is not the result of cell-fusion events. METHODS: We examined autopsy brain specimens from three sex-mismatched female bone-marrow transplantation patients, a female control, and a male control. We did immunohistochemistry, fluorescence in-situ hybridisation, and tissue analysis to look for multilineage, donor-derived neurogenesis. FINDINGS: Hippocampal cells containing a Y chromosome were present up to 6 years post-transplant in all three patients. Transgender neurons accounted for 1% of all neurons; there was no evidence of fusion events since only one X chromosome was present. Moreover, transgender astrocytes and microglia made up 1-2% of all glial cells. INTERPRETATION: Postnatal human neuropoiesis happens, and human haemopoietic cells can transdifferentiate into neurons, astrocytes, and microglia in a long term setting without fusing. Transplantable human haemopoietic cells could serve as a therapeutic source for long-term regenerative neuropoiesis. PMID- 15121407 TI - A non-infectious airborne disease. PMID- 15121408 TI - Treatment of severe acute graft-versus-host disease with third party haploidentical mesenchymal stem cells. AB - Adult bone-marrow-derived mesenchymal stem cells are immunosuppressive and prolong the rejection of mismatched skin grafts in animals. We transplanted haploidentical mesenchymal stem cells in a patient with severe treatment resistant grade IV acute graft-versus-host disease of the gut and liver. Clinical response was striking. The patient is now well after 1 year. We postulate that mesenchymal stem cells have a potent immunosuppressive effect in vivo. PMID- 15121409 TI - Immediate effects of n-3 fatty acid infusion on the induction of sustained ventricular tachycardia. AB - Increased consumption of n-3 fatty acids reduces mortality from sudden cardiac death, indicating that such acids have anti-arrhythmic effects. We did electrophysiological testing in ten patients with implanted cardioverter defibrillators who were at high risk of sudden cardiac death. To assess their immediate effects on the induction of sustained ventricular tachycardia, n-3 fatty acids were infused. Such tachycardia was not induced in five of seven patients. Our findings show that infusion of n-3 polyunsaturated fatty acids does not induce arrhythmia, but did result in a reduction of sustained ventricular tachycardia in some patients. PMID- 15121410 TI - Developmental dyslexia. AB - Developmental dyslexia, or specific reading disability, is a disorder in which children with normal intelligence and sensory abilities show learning deficits for reading. Substantial evidence has established its biological origin and the preponderance of phonological disorders even though important phenotypic variability and comorbidity have been recorded. Diverse theories have been proposed to account for the cognitive and neurological aspects of dyslexia. Findings of genetic studies show that different loci affect specific reading disability although a direct relation has not been established between symptoms and a given genomic locus. In both children and adults with dyslexia, results of neuroimaging studies suggest defective activity and abnormal connectivity between regions crucial for language functions--eg, the left fusiform gyrus for reading- and changes in brain activity associated with performance improvement after various remedial interventions. PMID- 15121411 TI - Portopulmonary hypertension and hepatopulmonary syndrome. AB - The clinically and pathophysiologically distinct entities of portopulmonary hypertension and hepatopulmonary syndrome occur in a substantial proportion of patients who have advanced liver disease of different causes. These disorders are notoriously underdiagnosed, but they have a substantial impact on survival and require focused treatment. Abnormal intrapulmonary vascular dilatation, the hallmark of hepatopulmonary syndrome, can cause profound hypoxaemia that can be very difficult to treat. By contrast, portopulmonary hypertension results from excessive pulmonary vasoconstriction and vascular remodelling that eventually leads to right-heart failure. Insights into the pathogeneses of these syndromes have led to novel therapeutic approaches. However, in severely affected patients, effective treatment remains a difficult task. In selected patients, liver transplantation represents the only treatment option, but the decision to do isolated liver transplantation is particularly challenging in patients who have severe pulmonary disease involvement. Data from several centres have contributed to provide criteria that allow improved prediction of which patients may, or may not, benefit from liver transplantation alone. PMID- 15121412 TI - Responding to the global human resources crisis. AB - The global community is in the midst of a growing response to health crises in developing countries, which is focused on mobilising financial resources and increasing access to essential medicines. However, the response has yet to tackle the most important aspect of health-care systems--the people that make them work. Human resources for health--the personnel that deliver public-health, clinical, and environmental services--are in disarray and decline in much of the developing world, particularly in sub-Saharan Africa. The reasons behind this disorder are complex. For decades, efforts have focused on building training institutions. What is becoming increasingly clear, however, is that issues of supply, demand, and mobility (transnational, regional, and local) are central to the human resource problem. Without substantial improvements in workforces, newly mobilised funds and commodities will not deliver on their promise. The global community needs to engage in four core strategies: raise the profile of the issue of human resources; improve the conceptual base and statistical evidence available to decision makers; collect, share, and learn from country experiences; and begin to formulate and enact policies at the country level that affect all aspects of the crisis. PMID- 15121413 TI - The lessons of MMR. PMID- 15121414 TI - Abuse of health-care workers' neutral status. PMID- 15121415 TI - A prisoner with acute renal failure. PMID- 15121416 TI - Unprotected sex and alloimmune activation. PMID- 15121418 TI - Quality of medical education in Mexico. PMID- 15121419 TI - Stopping HABITS. PMID- 15121420 TI - Stopping HABITS. PMID- 15121421 TI - Value of SERMs in postmenopausal women. PMID- 15121423 TI - More inequity in the UK National Health Service. PMID- 15121424 TI - Secondary crisis in African health care. PMID- 15121426 TI - Patients in medical education. PMID- 15121427 TI - Clinic. PMID- 15121428 TI - Reframing. PMID- 15121430 TI - Conflict(s) of interest. PMID- 15121431 TI - Using social marketing to increase recruitment of pregnant smokers to smoking cessation service: a success story. AB - OBJECTIVES: To explore what it is like to be a pregnant smoker in Sunderland and to inform the development of a Smoking Cessation Programme; to use qualitative techniques to develop a cessation programme tailored to pregnant smokers. STUDY DESIGN: The intervention follows years of social marketing research and development in Sunderland, Wearside, UK. METHODS: Information derived from nine focus groups (mainly with women from deprived areas, social class C2D and E) provided insights into the issues facing smoking pregnant women. This information was then used to overcome barriers to smoking cessation using the principles of social marketing. The number of women recruited into a specially designed smoking cessation support initiative was compared with women recruited into comparable groups in the North East. RESULTS: Recruitment of pregnant (and non-pregnant) smokers to the new NHS smoking cessation programme in Sunderland has increased during the intervention phase compared with neighbouring Primary Care Trust areas (in which different smoking cessation interventions targeted at pregnant women were being undertaken). CONCLUSIONS: This innovative intervention has been successful in generating ideas, guiding development of a customer-friendly service and encouraging women to come forward for smoking cessation support during their pregnancy. The target population have welcomed the approach, and health professionals have enjoyed and benefited from the role play with professional actors. PMID- 15121432 TI - Low-birthweight babies in Eastern Nigeria. PMID- 15121433 TI - Why do primary school children smoke? A longitudinal analysis of predictors of smoking uptake during pre-adolescence. AB - The objectives of this longitudinal study were to determine the prevalence of smoking among primary school children in Liverpool, and to identify the predictors of experimentation with cigarettes during pre-adolescence. A cohort of children (n = 270) completed questionnaires that elicited patterns of child smoking behaviour and children's experiences of smoking in their families and communities each year between the ages of 9 and 11 years. Parents also completed questionnaires. Children's first trials with cigarettes and repeated smoking were reported. The independent variables measured were socio-economic status, familial and peer smoking, and intentions to smoke. By age 11, 27% of children had tried smoking, 12% had smoked repeatedly and 3% were smoking regularly. Variables measured at age 9 predicting experimentation with cigarettes by age 11 were male gender 9P = 0.041) paternal smoking (P = 0.001) fraternal smoking (P = 0.017) a best friend who smoked (P = 0.026) and knowing someone with a smoking-related disease (P = 0.006) Intentions to smoke at age 9 did not predict smoking at age 11 (P < 0.001). In univariate analyses, child smoking was also associated with maternal smoking (P = 0.002 at age 11), living in a low-income household (P < 0.001 at age 10) and living in a deprived area ( P = 0.025 at age 11). Early smoking presents a considerable challenge to health promoters, not least because it is socially patterned. The interventions required must tackle the structural and social pressures that shape smoking behaviour during childhood. PMID- 15121434 TI - Epidemiological association between cigarette smoking and primary open-angle glaucoma: a meta-analysis. AB - OBJECTIVE: The role of cigarette smoking as a risk factor for primary open-angle glaucoma (POAG) remains controversial. The aim of this study was to examine this association by conducting a detailed meta-analysis of studies published in peer reviewed literature on the subject. METHODS: A comprehensive search for articles published up 31 December 2002 was performed, reviews of each study were conducted, and data were abstracted. Prior to meta-analysis, the studies were evaluated for publication bias and heterogeneity. RESULTS: Our analysis was based on seven reports. We found no evidence of publication bias or heterogeneity among the studies. The summary odds ratios from a fixed-effects model were 1.37 (95% CI: 1.00-1.87) for current smokers and 1.03 (95% CI: 0.77-1.38) for past smokers. CONCLUSIONS: Our meta-analysis results suggest that current smokers are at significantly increased risk of developing POAG. Efforts should be directed towards augmenting the campaign against smoking by adding the increased risk of POAG to the better-known arguments against smoking. PMID- 15121435 TI - Are support and parenting programmes of value for teenage parents? Who should provide them and what are the main goals? AB - OBJECTIVE: To review the evidence on what works in teenage parent support programmes; to determine the key elements of successful teenage parent support and parenting programmes; and to determine the gaps in the evidence. METHODS: Databases were searched, using a specific search strategy, for systematic reviews and randomised controlled trials, to ascertain whether social support, parenting programmes, or both, are effective in improving maternal or infant outcomes. The findings were summarized. RESULTS: Only one systematic review specifically addressed interventions among teenage parents and their children, although several randomised trials have since been published. CONCLUSIONS: Key questions remain for future support and parenting programmes. Social support and parenting interventions improve maternal-child interactions and child cognitive development, but do not reduce low-birth weight, stillbirth or neonatal death. Social support and parenting programmes need to be combined with measures to increase the minimum income, reduce smoking in pregnancy and increase breast feeding rates. Robust evaluations of packages of care in the UK are needed to inform national and local teenage pregnancy strategies. PMID- 15121436 TI - Changing patterns of adult (45-74 years) neurological deaths in the major Western world countries 1979-1997. AB - OBJECTIVES: To compare changes in 'adult' (45-74 years) 'all-cause deaths' (ACDs) with all neurological death categories by age and gender in the 10 major Western countries between the 1970s (1979-1981) and the 1990s (1995-1997). METHOD: World Health Organization standardized mortality data for age and gender (1979/97) were used to examine changes in adult mortality rates per million based upon ICD-9 categories for ACDs, 'neurological deaths' and the special neurological categories of 'other neurological deaths' (ONDs) and 'mental disorder deaths' (MDDs), which include the dementias. Ratios of ratios were calculated to demonstrate how each individual country's pattern changed over the period by age and gender, resolving the problem of cross-national comparisons. Rates of change across the endpoints and between age groups (45-54, 55-64, 65-74 and 75+ years) were examined using analysis of variance, stepwise regression analysis and cross tabulation analyses. RESULTS: Meningitis deaths fell substantially, but there was little change in multiple sclerosis or epilepsy deaths. OND rates for the 1990s increased compared with the 1970s rates for males and female, in actual terms and relative to ACDs for almost all countries. Many of the relative rates of increase were substantially higher than 20%. There were significant statistical differences with respect to relative rates of ONDs between the 1970s and the 1990s data, even when the 75+ years age group was excluded. Significant differences were also found between age groups, but only in the 1990s data. MDD rates showed similar trends. Analyses of actual rates of increase in these causes of death showed that males outnumber females in all ages below 74 years. The extent of this difference remained constant across the endpoints. However, in those aged 75 years and over, females outnumbered males at both endpoints, but this disparity widened significantly in the 1990s data. CONCLUSIONS: The 1990s data indicate substantial increases compared with the 1970s data for ONDs (especially amongst 65-74 year olds), and rises in MDDs in 55-64 year olds in five countries, including England and Wales and Germany, and in 65-74 year olds in most countries, suggesting earlier onsets of the underlying conditions. Further country-specific research is required to explain the emerging morbidity and mortality. PMID- 15121437 TI - Inappropriate hospital use by patients needing urgent medical attention in Italy. AB - The purpose of this study was to determine the extent of inappropriate admissions and days of stay in acute beds of patients who were admitted to hospital after attending the emergency department for urgent medical attention in Italy. The medical records of all adult patients (aged 16 years and over) on one randomly preselected day during the period January-December 2001 were reviewed. The retrospective application was made using the Appropriateness Evaluation Protocol list of criteria. Overall, 28.4 and 75.7% of hospital admissions and days of stay, respectively, were judged to be inappropriate. Results of the multiple logistic regression analysis indicated that inappropriate admission was significantly more likely in patients who lived closer to hospital, in younger patients, in patients from lower socio-economic classes, and in patients who did not receive diagnostic procedures in the emergency department. The proportion of patients whose admission was considered inappropriate was significantly lower in medical specialties wards. The number of inappropriate hospitalisation days was significantly higher in younger patients, in those admitted inappropriately, and in patients sampled close to discharge (during the final-third of his/her stay). The number of inappropriate hospitalisation days was significantly lower in patients admitted to surgical and traumatology/orthopaedics and medical specialties wards. Our findings indicate the need for specific interventions to reduce the prevalence of inappropriate hospital use and to modify physicians' attitudes and behaviours. PMID- 15121438 TI - Kohl (surma): a toxic traditional eye cosmetic study in Saudi Arabia. AB - The use of kohl (surma) as eyeliner is a popular practice in Saudi Arabia and people firmly believe that it is safe to use. A total of 107 kohl samples (branded and unbranded) were collected from different regions of Saudi Arabia, and analysed for the presence of lead. In addition, aluminium and antimony levels were also determined. Lead levels up to 53% were detected in some kohl preparations, and some samples were found to contain camphor and menthol. The blood analyses of regular kohl users revealed a high lead concentration and relatively low haemoglobin levels. Due to the health risk, an official public awareness campaign is suggested to encourage the use of lead-free kohl. PMID- 15121439 TI - Seroprevalence of common transfusion-transmitted infections among blood donors in Bangladesh. PMID- 15121440 TI - Comments on 'Palestinian refugee conditions associated with intestinal parasites and diarrhoea: Nuseirat refugee camp as a case study' by T.A. Abu Mourad (Public Health 2004; 118:131-42). PMID- 15121442 TI - Editorial comment on the correspondence from Rosen and Steinberg and Abu Mourad. PMID- 15121443 TI - Comments on "the effects of a 12-week group exercise programme on physiological and psychological variables and function in overweight women" by Grant et al. (Public Health 2004; 118:31-42). PMID- 15121444 TI - Evaluation of forearm microvascular blood flow regulation by laser Doppler flowmetry, iontophoresis, and curve analysis: contribution of axon reflex. AB - The relative contribution of vasodilating factors to the control of blood flow in the forearm cutaneous microcirculation is not well defined. Therefore, a mathematical transformation is introduced to decompose the superimposed signal and to investigate the involved mechanisms separately. Transdermal iontophoresis was used for the delivery of acetylcholine (ACh) or sodium nitroprusside (SNP) into the forearm, and cutaneous perfusion was measured using a laser Doppler flowmeter (LDF). The curve fitting procedure used in this study indicates that the LDF signal in response to ACh iontophoresis can be described by the superposition of two independent hyperbolic response curves. Obviously, each component of LDF signal indicates the existence of a separate mechanism, with corresponding rate constant k, latency T, and the saturation level Fmax. Blockade of C-fiber function (axon reflex) with topical anesthesia removes one of the two components of this response and allows the precise quantification of its contribution. SNP-evoked response also has two components, but their parameters were different from those of ACh. Therefore, ACh and SNP cause vasodilation in the skin microcirculation through different pathways. These findings have implications for clinical studies that use the iontophoresis technique for assessing vascular function and comparing responses to ACh and SNP to evaluate endothelial dysfunction. PMID- 15121446 TI - Microvessel density is not crucial for scintigraphic visualization of brain tumors using 99mTc-MIBI. AB - Functional imaging of brain tumors assists biopsy localization, therapy monitoring, and differentiating tumor recurrence from radiation necrosis. Tumor vascularization is a strong prognostic predictor in solid tumors and also a key factor of tracer uptake. However, the relationship of brain tumor vascularization and functional imaging has not yet been investigated sufficiently so far. In the present study, we correlated histologically assessed microvessel density as an objective parameter for brain tumor vascularization with imaging data. Four male patients were studied. After 99mTc-MIBI scintigraphy, all patients had a MRI within 2 weeks. Histology showed microcystic astrocytoma, glioblastoma (n = 2), and anaplastic oligodendroglioma, respectively. Microvessel density was lowest in the microcystic astrocytoma, medium in the glioblastomas, and highest in the anaplastic oligodendroglioma. Scintigraphy visualized only the glioblastomas, but not the microcystic astrocytoma or oligodendroglioma. Our data showed no correlation between tumor microvessel density and 99mTc-MIBI scintigraphy. Thus, we conclude that scintigraphic visualization of brain tumors is not strictly dependent on tumor vascularization. PMID- 15121445 TI - A method for selectively limiting lumen diameter in corrosion casting. AB - Corrosion casting is a technique frequently used to evaluate the form and spatial relationship of three dimensional biological structures, such as vascular networks, in vitro [Scann. Microsc. 5 (1991) 1097; Schraufnagel, D.E. The lung microstructure. In: Motta, P.M., Murakami, T. Fujita, H. eds. Scanning Electron Microscopy of Vascular Casts: Methods and Applications. Boston: Kluwer Academic Publishers; 1992:123-137]. However, because corrosion casts tend to reproduce the complexity of surrounding vascular structures as well as the structures of interest, the use of this technique in highly complex vascular systems may obscure important changes such as A-V shunts because they may get lost among the myriad of capillaries. We developed a novel modification for creating vascular corrosion casts by utilizing polystyrene microspheres to selectively embolize normal capillary networks and thereby reduce the overall complexity of the cast. his technique may aid investigators in the evaluation of a variety of vascular beds and is useful in demonstrating non-capillary arteriovenous communications. PMID- 15121447 TI - A graph theory analysis of renal glomerular microvascular networks. AB - A graph theory model and its invariants are used to compare previously published renal glomerular networks of six adult rats, one adult uremic rat, and one newborn rat. Invariants calculated include order, size, cycle rank, eccentricity, root distance, planarity, and vertex degree distribution. These invariants enabled the differentiation of six normal adult glomerular microvascular networks from that of the uremic glomerulus and from that of the normal newborn glomerulus. These invariants might then be used to differentiate between normal and pathological vascular networks. Also proposed are graph theory invariants that might be used to develop a quantitative model for angiogenesis. PMID- 15121448 TI - Measurement of macromolecular diffusion coefficients in human tumors. AB - The diffusive transport of macromolecules in tumors is an important determinant of the delivery of many anticancer therapeutics. However, measurements of diffusive transport to date have only been performed in animal models. In this work, diffusion coefficients of BSA and IgM were measured in human tumor biopsies (cooled to 4-7 degrees C to prevent degradation) using fluorescence recovery after photobleaching. To quantify the effects of excision and cooling, the diffusion coefficient of BSA and IgM was measured in human tumor xenografts in situ and after cooling and excision. The change in diffusion coefficients before and after excision of xenografts was used to calculate in vivo diffusion coefficients in human tumors from ex vivo measurements. Using this approach, we obtained the first quantitative determinations of macromolecular diffusion coefficients in human tumors and find that the diffusion coefficients of BSA and IgM in human colon were adenocarcinomas higher than those in xenografts. This difference is consistent with lower collagen content in the accessible regions of these human tumors. These measurements allow the quantitative prediction of the diffusive transport of like-sized macromolecular therapeutics in human tumors. These measurements should help in modeling the transport of novel large MW therapeutics, and hence in estimating their distribution and efficacy in tumors. PMID- 15121450 TI - Diapause induction and clock mechanism in the cabbage beetle, Colaphellus bowringi (Coleoptera: Chrysomelidae). AB - Photoperiodic control of diapause induction was investigated in the short-day species, Colaphellus bowringi, which enters summer and winter diapause as adult in the soil. Photoperiodic responses at 25 and 28 degrees C revealed a critical night length between 10 and 12 h; night lengths > or =12 h prevented diapause, whereas night lengths <12 h induced summer diapause in different degree. Experiments using non-24-h light-dark cycles showed that the duration of scotophase played an essential role in the determination of diapause. Night interruption experiments with T=24 h showed that diapause was effectively induced by a 2-h light pulse in most scotophases; whereas day-interruption experiments by a 2-h dark break had a little effect on the incidence of diapause. The experiments of alternating short-night cycles (LD 16:8) and long-night cycles (LD 12:12) during the sensitive larval period showed that the information of short nights as well as long nights could be accumulated. Nanda-Hamner experiments showed three declining peaks of diapause at 24 h circadian intervals. Bunsow experiments showed two very weak peaks for diapause induction, one being 8 h after lights-off, and another 8 h before lights-on, but it did not show peaks of diapause at a 24 h interval. These results suggest that the circadian oscillatory system constitutes a part of the photoperiodic clock of this beetle but plays a limited role in its photoperiodic time measurement. PMID- 15121449 TI - A decrease in effective diameter of rat mesenteric venules due to leukocyte margination after a bolus injection of pentoxifylline--digital image analysis of an intravital microscopic observation. AB - The ability of leukocytes to adhere to endothelial cells (EC) and then to migrate out of the blood stream into tissues enable them to perform their surveillance functions. Adhesion of leukocytes to EC is, however, only possible if the cells have marginated as a result of rheological interaction with other blood cells in flow. Using Pentoxifylline (PTX), a rheologically active drug, to manipulate this interaction, we have imaged and quantified this margination phenomenon in vivo. A system has been developing to perform this imaging via an intravital microscope connected to an image processing system. Albino rats were anesthetized and cannulated for intravenous bolus injection (0.5 ml) of PTX (1.25 mg/ml) through the femoral vein. A longitudinal incision exposed the mesentery, part of which was observed under microscope to visualize microcirculation. The image of interest was then stored on computer hard drive. Individual leukocyte velocities were determined before and after PTX infusion. The leukocytes, marginating and sticking after PTX infusion either remained attached, constituting the peripheral marginating leukocyte pool in the postcapillary venules, or detached with different step velocities. The reduction in effective venular diameters as a result of leukocyte margination was estimated to be 32-44%. These results demonstrate the biological importance of hemodynamic displacement leading to docking, adhesion, rolling and migration processes of leukocytes in blood. PMID- 15121451 TI - Nutrition, hormones and life history in burying beetles. AB - Nutrition, hormones and the allocation of physiological resources are intricately related. To investigate these inter-relationships in female burying beetles (Nicrophorus spp.), we examined the effect of diet quality on juvenile hormone (JH) levels and reproduction, and the effect of JH supplementation on reproduction and resistance to starvation. Nicrophorus orbicollis adult females fed a less preferred mealworm larvae diet gained less body mass, had smaller ovaries and had lower titers of JH in their hemolymph than females fed a preferred blowfly diet. When presented a carcass for breeding, females on a less preferred diet oviposited 33% fewer eggs, and eggs were of 18% less mass. Females on the less preferred diet also took longer to begin oviposition as indicated indirectly by the time when their eggs hatched. To investigate the effects of JH, independent of nutrition, JH was topically applied to single and paired females of Nicrophorus tomentosus. When presented a carcass, JH-treated paired females oviposited more eggs (28%-year 1, 44%-year 2) than control females, and also showed a trend toward faster oviposition. JH supplementation had a greater effect on single females. JH treatment increased the proportion of single females attempting reproduction (at least one viable larva), increased the number of eggs (69%-year 1, 123%-year 2), and increased the proportion of females ovipositing early. In separate experiments, treatment with JH or a JH analog negatively affected resistance to starvation in three species. Treatment with JH reduced starvation survival by 10.3% days in N. tomentosus females. Treatment with the JH analog methoprene reduced starvation survival 17.8% in N. orbicollis females and by 18% in Ptomascopus morio females. These results suggest that JH has positive and negative effects on different components of life history. PMID- 15121452 TI - Green colouration of cocoons in Antheraea yamamai (Lepidoptera: Saturniidae): light-induced production of blue bilin in the larval haemolymph. AB - When the larvae of a saturniid silkmoth, Antheraea yamamai, are maintained under high intensity light (5000 lux), they produce green cocoons whereas the cocoons produced under light of low intensity (e.g., 50 lux) or in darkness are yellow. The green colour of the cocoon is due to the presence of a blue bilin pigment in combination with yellow pigment, and light stimulates the accumulation of blue bilin. In the present study, we show that two blue bilins, with similar characteristics to the sarpedobilin in the green cocoon, can be induced in larval haemolymph both in vivo and in vitro. In both conditions, the amount of these bilins increased with increasing intensity or duration of light exposure. Induction also occurred at 0 degrees C. In contrast, the chromophore of the constitutive biliprotein of the haemolymph did not change depending on light conditions. Size fractionation of the haemolymph indicates that the precursor of the blue bilins induced by light is bound to a protein with a molecular mass of 5000 Da or more. Thus, in these insects, the blue bilin responsible for green colouration is facultative under photochemical stimulation. PMID- 15121453 TI - Intra-specific variation in female remating in Callosobruchus chinensis and C. maculatus. AB - The effects of mating duration on female remating (exp. 1) and under different male densities (exp. 2) were examined in two strains of the adzuki bean beetle, Callosobruchus chinensis and in one strain of the bruchid beetle, C. maculatus. In experiment 1, the frequency of female remating was markedly different between the two strains of C. chinensis. Females of the jC strain, reared long-term in the laboratory, did not remate after being allowed to mate freely (=monogamy), whereas females of the isC strain, recently established from the field, showed high remating frequencies (=polyandry). In both strains, the frequency of female remating increased after the duration of the first mating was deliberately shortened. The relation between mating duration and remating frequency was significantly different, however, between the two strains. In a closely related species, C. maculatus, which manifests polyandry, this relation was more similar to that of the field-derived (=isC) than to that of the laboratory-derived (=jC) strain of C. chinensis. The reasons for the inter-strain variation observed in the remating frequencies of C. chinensis are also discussed. In experiment 2, the mating duration of the three strains was compared under different male densities. Only the lab-derived strain demonstrated a significantly shorter mating duration when one female was placed together with five males than when paired with one male. The shorter mating duration (approximately 26 s) was similar to that of females allowed to remate in the monogamous strain in experiment 1. PMID- 15121454 TI - Induced hyperlipaemia and immune challenge in locusts. AB - Injections of immunogens, such as beta-1,3-glucan or lipopolysaccharide (LPS), bring about a marked hyperlipaemia with associated changes in lipophorins and apolipophorin-III in the haemolymph of Locusta migratoria. These changes are similar to those observed after injection of adipokinetic hormone (AKH). The possibility that endogenous AKH is released as part of the response to these immunogens is investigated using passive immunisation against AKH-I, and measurement of AKH-I titre in the haemolymph after injection of immunogens. The data presented show that, despite the similarity of the changes brought about by the presence of immunogens in the haemolymph to those brought about by AKH, there is no release of endogenous AKH after injection of laminarin or LPS. A direct effect of the immunogens on release of neutral lipids by the fat body cannot be demonstrated in vitro, and the mechanism by which hyperlipaemia is induced during immune challenge remains uncertain. PMID- 15121455 TI - Metabolic rate variation in Glossina pallidipes (Diptera: Glossinidae): gender, ageing and repeatability. AB - Despite the importance of metabolic rate in determining flight time of tsetse and in mediating the influence of abiotic variables on life history parameters (and hence abundance and distribution), metabolic rate measurements and their repeatability have not been widely assessed in these flies. We investigate age related changes in standard metabolic rate (SMR) and its repeatability, using flow-through respirometry, for a variety of feeding, gender and pregnancy classes during early adult development in laboratory-reared individuals of the tsetse fly, Glossina pallidipes. Standard metabolic rate (144-635 microW) was generally within 22% of previous estimates, though lower than the values found using closed system respirometry. There was no significant difference between the genders, but metabolic rate increased consistently with age, probably owing to flight muscle development. Repeatability of metabolic rate was generally high (r=0.6-.09), but not in younger teneral adults and pregnant females (r approximately equal to 0.05 0.4). In these individuals, low repeatability values are a consequence of muscle or in utero larval development. Tsetse and other flies generally have a much higher metabolic rate, for a given size, than do other insect species investigated to date. PMID- 15121456 TI - Dietary glycerol and adult access to water: effects on fecundity and longevity in the almond moth. AB - The quality of food eaten by larval insects will affect traits such as gamete production, fat reserves, muscle bulk and body size in the adult. Moreover, larvae also depend on high moisture content in the diet for survival. The almond moth (Ephestia cautella) (W.) (Lepidoptera; Pyralidae) does not feed as an adult although it continues to drink water. We tested the idea that an almond moth could compensate for a low-water diet as a larva by increasing its water intake as an adult. We reared larvae on two different food sources with different moisture regimes; standard laboratory diet with glycerol (relatively wet) and standard diet without glycerol (relatively dry). Half the adult moths from each treatment were given water to drink before their first and only mating. Our results show that wet larval diets (i.e. containing glycerol) significantly decreased fecundity (total number of eggs laid and the proportion of hatched larvae), whilst it significantly increased male and female longevity. The interaction effect of water access for adult males and females was significant, independent of the glycerol in the larval diet. Longevity in females that were not presented with water as adults was slightly higher if mated with a male that had had access to water, suggesting a mating donation of water. However, females that received water as adults showed a decreased longevity if mated with a male who had also had access to water as an adult, indicating a negative effect of water if received by both males and females. In addition, when the larval diet included glycerol, increased number of eggs laid decreased female longevity, whilst an absence of glycerol in the larval diet resulted in low female longevity that was unlinked with fecundity. Glycerol is used in many artificial insect diets and the fact that it shows a strong effect on key life-history traits (reproductive output and longevity in this species), merits careful re examination of its effects on these important traits in other laboratory models. We also discuss the possibility that larval diet can affect female reproductive decisions. PMID- 15121457 TI - Effects of Bt plants on the development and survival of the parasitoid Cotesia plutellae (Hymenoptera: Braconidae) in susceptible and Bt-resistant larvae of the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae). AB - A range of crops have been transformed with delta-endotoxin genes from Bacillus thuringiensis (Bt) to produce transgenic plants with high levels of resistance to lepidopteran pests. Parasitoids are important natural enemies of lepidopteran larvae and the effects of Bt plants on these non-target insects have to be investigated to avoid unnecessary disruption of biological control. This study investigated the effects of Cry1Ac-expressing transgenic oilseed rape (Brassica napus) on the solitary braconid endoparasitoid Cotesia plutellae in small-scale laboratory experiments. C. plutellae is an important natural enemy of the diamondback moth (Plutella xylostella), the most important pest of brassica crops world-wide. Bt oilseed rape caused 100% mortality of a Bt-susceptible P. xylostella strain but no mortality of the Bt-resistant P. xylostella strain NO QA. C. plutellae eggs laid in Bt-susceptible hosts feeding on Bt leaves hatched but premature host mortality did not allow C. plutellae larvae to complete their development. In contrast, C. plutellae developed to maturity in Bt-resistant hosts fed on Bt oilseed rape leaves and there was no effect of Bt plants on percentage parasitism, time to emergence from hosts, time to adult emergence and percentage adult emergence from cocoons. Weights of female progeny after development in Bt-resistant hosts did not differ between plant types but male progeny was significantly heavier on wildtype plants in one of two experiments. The proportion of female progeny was significantly higher on Bt plants in the first experiment with Bt-resistant hosts but this effect was not observed again when the experiment was repeated. PMID- 15121458 TI - Lack of correlation between body mass and metabolic rate in Drosophila melanogaster. AB - We examined the association between body mass and metabolic rate in Drosophila melanogaster under a variety of conditions. These included comparisons of body mass and metabolic rate in flies from different laboratory lines measured at different ages, over different metabolic sampling periods, and comparisons using wet versus dry mass data. In addition, the relationship between body mass and metabolic rate was determined for flies recently collected from wild populations. In no case was there a significant correlation between body mass and metabolic rate. These results indicate that care must be taken when attempting to account for the effects of body mass on metabolic rate. Expressing such data in mass specific units may be an inappropriate method of attempting to control for the effects of differences in body mass. PMID- 15121459 TI - Continuous recording of excretory water loss from Musca domestica using a flow through humidity meter: hormonal control of diuresis. AB - Water loss from adult male houseflies was continuously recorded using a flow through humidity meter, which enabled losses to be apportioned between the sum of cuticular and respiratory transpiration, salivation and excretion. Transpiration accounted for >95% of water lost from sham-injected flies, compared with excretion (3.0%) and salivation (2.4%). In contrast, excretion accounted for 40% of water lost from flies injected with > or =3 microl of saline, whereas salivary losses were unchanged. Saline injections (1-5 microl) expanded the abdomen in the dorsal-ventral plane, and this expansion was positively correlated with the magnitude of the ensuing diuresis, suggesting the signal for diuretic hormone release originates from stretch receptors in abdominal tergal-sternal muscles. The effects of decapitation, severing the ventral nerve cord within the neck or ligaturing the neck, showed the head was needed to initiate and maintain diuresis, but was neither the source of diuretic hormone nor did it control the discharge of urine from the anus. These findings indicate the head is part of the neural-endocrine pathway between abdominal stretch receptors and sites for diuretic hormone release from the thoracic-abdominal ganglion mass. Evidence is presented for Musdo-K having a hormonal role in the control of diuresis, although other neuropeptides may also be implicated. PMID- 15121460 TI - IFFS Surveillance 04. PMID- 15121478 TI - The origin of schizophrenia: genetic thesis, epigenetic antithesis, and resolving synthesis. AB - Traditionally, it has been thought that schizophrenia results from the interaction of predisposing genes and hazardous environmental factors. In this article, the paradigm of "genes plus environment" is challenged, and a new interpretation is presented, in which the emphasis on DNA sequence variation is shared with epigenetic misregulation as a critical etiopathogenic factor. Partial epigenetic stability (metastability) of gene regulation is consistent with various nonmendelian irregularities of schizophrenia, such as the presence of clinically indistinguishable sporadic and familial cases, discordance of monozygotic twins, coincidence of peaks of susceptibility with major endocrine rearrangements, and fluctuating course of disease severity, among others. It is also suggested that stochastic epigenetic events might account for a substantial portion of phenotypic variance, which traditionally has been ascribed to environmental effects. This theoretic essay is constructed according to the principle of Hegelian dialectic reasoning (thesis-antithesis-synthesis), which serves the goal of showing that the best outcome of molecular genetic studies in schizophrenia (and perhaps other complex diseases) can be expected when components that effect chromatin structure and gene regulation are taken into account and investigated comprehensively. PMID- 15121479 TI - Strong evidence for association between the dystrobrevin binding protein 1 gene (DTNBP1) and schizophrenia in 488 parent-offspring trios from Bulgaria. AB - BACKGROUND: The gene encoding the dystrobrevin binding protein (DTNBP1) has been implicated in the pathogenesis of schizophrenia by several association studies. We tried to replicate these findings in a sample of 488 parent-proband trios recruited in Bulgaria. Probands had a diagnosis of schizophrenia (n = 441) or schizoaffective disorder (n = 47). METHODS: We genotyped eight single nucleotide polymorphisms within the gene, four of which had been reported in previous studies, and four identified as informative by our group through direct screening of the gene and genotyping in a sample of cases and control subjects. RESULTS: A significant excess of transmissions was observed for two of the markers, p1635 and p1757, (p =.0009 and.0013, respectively). Analysis of two-, three-, and four marker haplotypes produced numerous positive results, with six (4% of the total combinations) at p <.001. CONCLUSIONS: These results provide strong support for DTNBP1 as a susceptibility gene for schizophrenia; however, different haplotypes seem to be associated in different studies. PMID- 15121480 TI - Polymorphisms in the 13q33.2 gene G72/G30 are associated with childhood-onset schizophrenia and psychosis not otherwise specified. AB - BACKGROUND: Childhood-onset schizophrenia (COS), defined as onset of psychotic symptoms by age 12 years, is a rare and severe form of the disorder that seems to be clinically and neurobiologically continuous with the adult disorder. METHODS: We studied a rare cohort consisting of 98 probands; 71 of these probands received a DSM-defined diagnosis of schizophrenia, and the remaining 27 were diagnosed as psychosis not otherwise specified (NOS) (upon 2-6 year follow-up, 13 have subsequently developed bipolar disorder). Two overlapping genes, G72 and G30 on 13q33.2, were identified through linkage-disequilibrium-based positional cloning. Single nucleotide polymorphisms (SNPs) at the G72/G30 locus were independently associated with both bipolar illness and schizophrenia. We analyzed SNPs at this locus with a family-based transmission disequilibrium test (TDT) and haplotype analyses for the discrete trait, as well as quantitative TDT for intermediate phenotypes, using the 88 probands (including COS and psychosis-NOS) with parental participation. RESULTS: We observed significant pairwise and haplotype associations between SNPs at the G72/G30 locus and psychotic illness. Furthermore, these markers showed associations with scores on a premorbid phenotype measured by the Autism Screening Questionnaire, and with age of onset. CONCLUSIONS: These findings, although limited by potential referral bias, confirm and strengthen previous reports that G72/G30 is a susceptibility locus both for schizophrenia and bipolar disorder. PMID- 15121481 TI - Identification of PIK3C3 promoter variant associated with bipolar disorder and schizophrenia. AB - BACKGROUND: Genes involved in phosphoinositide (PI) lipid metabolism are excellent candidates to consider in the pathogenesis of bipolar disorder (BD) and schizophrenia (SZ). One is PIK3C3, a member of the phosphatidylinositide 3-kinase family that maps closely to markers on 18q linked to both BD and SZ in a few studies. METHODS: The promoter region of PIK3C3 was analyzed for mutations by single-strand conformation polymorphism analysis and sequencing. A case-control association study was conducted to determine the distribution of variant alleles in unrelated patients from three cohorts. Electromobility gel shift assays (EMSA) were performed to assess the functional significance of variants. RESULTS: Two polymorphisms in complete linked disequilibrium with each other were identified, 432C- > T and a "C" insert at position -86. The -432T allele occurs within an octamer containing an ATTT motif resembling members of the POU family of transcription factors. In each population analyzed, an increase in -432T was found in patients. EMSAs showed that a -432T containing oligonucleotide binds to brain proteins that do not recognize -432C. CONCLUSIONS: A promoter mutation in a PI regulator affecting the binding of a POU-type transcription factor may be involved in BD and SZ in a subset of patients. PMID- 15121482 TI - Pervasive developmental disorder and childhood-onset schizophrenia: comorbid disorder or a phenotypic variant of a very early onset illness? AB - BACKGROUND: Childhood-onset schizophrenia (COS) is a severe form of the adult onset disorder with a high rate of premorbid developmental abnormalities. Early symptoms of pervasive developmental disorder (PDD) have been reported in five independent studies of COS. In this study, we compared evidence for premorbid PDD as a nonspecific manifestation of impaired neurodevelopment seen in schizophrenia, or as an independent risk factor for COS. METHODS: Diagnosis of past or current autism or PDD was made according to the DSM-IV criteria. COS patients with and without PDD were compared with respect to neuropsychological, clinical, and neurobiological measures. Several candidate genes for autism were examined in the entire COS sample and the subgroup with PDD using the Transmission Disequilibrium Test (TDT) and Quantitative TDT (QTDT). RESULTS: Nineteen (25%) of COS probands had a lifetime diagnosis of PDD: one met criteria for autism, two for Asperger's disorder, and 16 for PDD not otherwise specified. Premorbid social impairment was most common feature for COS-PDD subjects. The PDD group did not differ from the rest of the COS sample with respect to age of onset, IQ, response to medications, and rate of familial schizotypy. Unexpectedly, two siblings of COS-PDD probands met criteria for nuclear autism. There was no difference between PDD and non-PDD groups with respect to initial brain magnetic resonance imaging (MRI) measures. However, rate of gray matter loss was greater for PDD (n = 12) than for the non-PDD (n = 27) subgroup (-19.5 +/- 11.3 mL/year vs. -9.6 +/- 15.3 mL/year; p =.05). None of eight candidate genes for autism were associated with COS or COS-PDD. CONCLUSIONS: Premorbid PDD in COS is more likely to be a nonspecific marker of severe early abnormal neurodevelopment. However, the occurrence of two siblings of COS-PDD probands (17%) with nuclear autism remains to be understood. PMID- 15121483 TI - Sustained attention deficits in nonpsychotic relatives of schizophrenic patients: a recurrence risk ratio analysis. AB - BACKGROUND: In nonpsychotic parents and siblings of schizophrenic patients, the recurrence risk ratios of sustained attention deficits, as measured by the continuous performance test (CPT), were examined with a series of cut-off points. METHODS: Among 116 parents and 95 siblings of 91 schizophrenic probands in northern Taiwan, both undegraded and degraded sessions of the CPT were administered. Subjects' signal detection sensitivity of CPT performance (d') was standardized against a community sample without (unadjusted z score) or with (adjusted z score) adjustment for age, gender, and educational level. RESULTS: Differences in the risk ratios between the parents and siblings that were based on the unadjusted z scores of CPT d' diminished markedly if the adjusted z scores were used. As the cut-off point in the adjusted z score decreased from -2.5 to 3.0, the risk ratio increased continually for both the undegraded (10.1-18.8 for parents, 10.0-16.7 for siblings) and degraded (12.4-102.7 for parents, 8.6-72.0 for siblings) test. CONCLUSIONS: Stringent cut-off criteria of CPT deficits with adjustment for demographic features leads to recurrence risk ratios greater than those based on schizophrenia alone in both parents and siblings of schizophrenic patients. PMID- 15121484 TI - Striatal amphetamine-induced dopamine release in patients with schizotypal personality disorder studied with single photon emission computed tomography and [123I]iodobenzamide. AB - BACKGROUND: Previous imaging studies demonstrated that schizophrenia is associated with increased amphetamine-induced dopamine (DA) release in the striatum, most pronounced during episodes of illness exacerbation. Schizotypal personality disorder (SPD) is a schizophrenia spectrum disorder, genetically related to schizophrenia. The goal of this study was to investigate striatal DA function in patients with SPD. METHODS: In our study, 13 SPD patients and 13 matched healthy control subjects underwent single photon emission computed tomography (SPECT) scan during bolus plus constant infusion of the D2/3 radiotracer [123I]iodobenzamide (IBZM). Striatal specific to nonspecific equilibrium partition coefficient (V(3)") was measured at baseline and following amphetamine administration (.3 mg/kg). RESULTS: No significant differences were observed in baseline V(3)" between groups. Amphetamine induced a larger decrease in [123I]IBZM V(3)" in SPD patients (-12 +/- 5%) compared with control subjects ( 7 +/- 5%, p =.03). CONCLUSIONS: The reduction in [123I]IBZM V(3)" induced by amphetamine in SPD was similar to that observed in remitted schizophrenia patients (-10 +/- 9%, n = 17), but significantly lower than that observed during illness exacerbation (-24 +/- 13%, n = 17). This suggests that DA dysregulation in schizophrenia spectrum disorders might have a trait component, present in remitted patients with schizophrenia and in SPD, and a state component, associated with psychotic exacerbations but not SPD. PMID- 15121486 TI - A meta-analysis and critical review of the effects of conventional neuroleptic treatment on cognition in schizophrenia: opening a closed book. AB - BACKGROUND: In recent years, numerous studies have examined whether new generation antipsychotic agents impact cognitive impairment in patients with schizophrenia when compared with conventional antipsychotic treatment. The working assumption of such studies, supported by narrative reviews, is that the conventional antipsychotic medications have little or no effect on cognition. The studies concerning the effects of conventional neuroleptics on cognition, however, have never been analyzed quantitatively. In this meta-analysis, we revisit the question of whether typical agents might have enhancing effects on cognition. METHODS: The meta-analysis included studies that 1) compared the effects of typical neuroleptic agents with those of placebo or no medication on cognition; 2) examined adult patients identified as having schizophrenia; and 3) produced adequate data to estimate an effect size. Studies were identified by searches of computerized literature databases and by cross-referencing included studies. The effect size calculated was d, the difference between the means in cognitive measures of patients with schizophrenia taking and not taking conventional neuroleptic medication, divided by the pooled SD. Critically, when multiple measures were reported in a single article, effect sizes were combined so as to minimize the possibility that one study had undue weight simply because of the number of cognitive outcome variables that it contained. These results were corrected for bias due to sample size, with each of the effect sizes weighted by the reciprocal of its variance. The final sample after exclusion of outliers comprised 208 effect sizes from 34 studies. RESULTS: With a random effects model, effect sizes from the primary studies were weighted according to sample size and averaged. The resulting mean effect size was.22 (95% confidence interval =.10,.34). The result is positive, in that the range did not include zero and was of low moderate size. No moderating effects of study design or patient qualities were found to be significant. With the same procedures, effect sizes for individual neurocognitive domains were computed. Effect sizes were generally in the.13-.29 range for the majority of cognitive functions, whereas motor function was impacted negatively (-.11). Unexpectedly, medication dose did not correlate with effect size. CONCLUSIONS: Typical antipsychotic medication provides modest-to-moderate gains in multiple cognitive domains. Given unavoidable methodologic limitations of the primary studies, current findings suggest that the impact of conventional medication on cognitive function should be re-evaluated. PMID- 15121485 TI - Human positron emission tomography studies of brain neurokinin 1 receptor occupancy by aprepitant. AB - BACKGROUND: Aprepitant is a highly selective substance P (neurokinin 1 [NK(1)] receptor) antagonist that significantly improves the pharmacotherapy of acute and delayed highly emetogenic chemotherapy-induced nausea and vomiting, probably through an action in the brain stem region of the central nervous system. Here, we report the use of positron emission tomography imaging with the NK(1) receptor binding-selective tracer [(18)F]SPA-RQC to determine the levels of central NK(1) receptor occupancy achieved by therapeutically relevant doses of aprepitant in healthy humans. METHODS: Two single-blind, randomized, placebo-controlled studies in healthy subjects were performed. The first study evaluated the plasma concentration-occupancy relationships for aprepitant dosed orally at 10, 30, 100, or 300 mg, or placebo (n = 12). The second study similarly evaluated oral aprepitant 30 mg and placebo (n = 4). In each study, dosing was once daily for 14 consecutive days. Data from both studies were combined for analyses. The ratio of striatal/cerebellar [(18)F]SPA-RQ (high receptor density region/reference region lacking receptors) was used to calculate trough receptor occupancy 24 hours after the last dose of aprepitant. RESULTS: Brain NK(1) receptor occupancy increased after oral aprepitant dosing in both a plasma concentration-related (r =.97; 95% confidence interval [CI] =.94-1.00, p <.001) and a dose-related (r =.94; 95% CI =.86-1.00, p <.001) fashion. High (> or =90%) receptor occupancy was achieved at doses of 100 mg/day or greater. The plasma concentrations of aprepitant that achieved 50% and 90% occupancy were estimated as approximately 10 ng/mL and approximately 100 ng/mL, respectively. CONCLUSIONS: Positron emission tomography imaging with [(18)F]SPA-RQ allows brain NK(1) receptor occupancy by aprepitant to be predicted from plasma drug concentrations and can be used to guide dose selection for clinical trials of NK(1) receptor antagonists in central therapeutic indications. PMID- 15121487 TI - Association between serotonin transporter gene promoter polymorphism and suicide: results of a meta-analysis. AB - BACKGROUND: There is strong evidence supporting a role for serotonin system dysfunction in the pathology of suicidal behavior. Many studies have examined the association between a functional polymorphism of the serotonin transporter gene promoter (5-HTTLPR) and suicide but have yielded inconsistent results. Our goal here, by analyzing the cumulative data from primary literature, was to determine conclusively whether there is an association. METHODS: Three meta-analyses were performed. One compared the 5-HTTLPR polymorphism between suicidal subjects and normal control subjects; another compared suicide attempters with nonattempters of the same psychiatric diagnoses; the last one compared either violent or nonviolent suicidal subjects with normal control subjects. RESULTS: We found no association between 5-HTTLPR polymorphism and suicidal behavior (p =.379). When we compared subjects with the same psychiatric diagnoses, the genotypes carrying the s allele were significantly more frequent in suicide attempters than in nonattempters (p =.004). In addition, the s allele was associated with violent suicide (p =.0001) but not with nonviolent suicide (p = 1.00). CONCLUSIONS: Our results provide significant evidence supporting the association of the s allele of 5-HTTLPR polymorphism with suicidal behavior in the psychiatric population, also with violent suicide. These support a role for decreased serotonin transporter function in the vulnerability to suicide in a select population. PMID- 15121488 TI - Modafinil improves cognition and response inhibition in adult attention deficit/hyperactivity disorder. AB - BACKGROUND: Modafinil, a novel cognitive enhancer, has a clinical profile similar to conventional stimulants such as methylphenidate, despite a seemingly different mechanism of action. Modafinil selectively improves neuropsychological task performance in healthy volunteers, possibly through improved inhibitory control. We examined whether modafinil induced similar improvements in adults with attention-deficit/hyperactivity disorder. METHODS: Twenty patients with a DSM-IV diagnosis of attention-deficit/hyperactivity disorder were entered into a double blind, randomized, placebo-controlled crossover study using a single 200 mg dose of modafinil. RESULTS: Modafinil produced a similar pattern of cognitive enhancement to that observed in healthy adults, with improvements on tests of short-term memory span, visual memory, spatial planning, and stop-signal motor inhibition. On several measures, increased accuracy was accompanied by slowed response latency. This alteration in the speed-accuracy trade-off may indicate that modafinil increases the ability to "reflect" on problems coupled with decreased impulsive responding. Improvements were also seen in sustained attention, which was unaffected in healthy subjects. CONCLUSIONS: If these benefits are shown to be maintained with chronic administration, modafinil may have potential as an important therapy for attention-deficit/hyperactivity disorder with a similar effect to stimulants such as methylphenidate in improving stop-signal response inhibition but without the side effects commonly experienced with amphetamine-like drugs. PMID- 15121489 TI - Low level of dopaminergic D2 receptor binding in obsessive-compulsive disorder. AB - BACKGROUND: Despite growing evidence for involvement of the dopaminergic system in obsessive-compulsive disorder (OCD), the functional anatomy of the dopaminergic system in the basal ganglia has been investigated sparsely. METHODS: Dopamine D(2) receptor binding was assessed in 10 medication-free OCD patients and 10 healthy control subjects, matched for age, gender, and handedness. The binding potential was measured with single photon emission computerized tomography (SPECT) and infusion of the D(2) receptor radiotracer [(123)I] iodobenzamide. With magnetic resonance imaging as reference, regions of interest (caudate and putamen) were delineated for each hemisphere and coregistered with the corresponding SPECT scans. RESULTS: Dopamine D(2) receptor binding in the left caudate nucleus was significantly lower in the patients with OCD than in healthy control subjects [F(1,18) = 7.0, p =.016]. In addition, an interhemispheric difference was observed in the patient sample. Both the D(2) receptor binding potential (df = 9, p =.012), and the volume (df = 9, p =.029) of the left caudate nucleus were statistically significantly reduced relative to the right caudate nucleus. CONCLUSIONS: This study provides in vivo evidence for abnormalities in the binding potential of the dopamine D(2) receptor, which suggest the direct involvement of the dopaminergic system in the pathophysiology of OCD. PMID- 15121490 TI - Role of dietary vitamin K intake in chronic oral anticoagulation: prospective evidence from observational and randomized protocols. AB - PURPOSE: The potential association between vitamin K intake and coagulation instability has been explored primarily in case reports and small, retrospective, uncontrolled studies. We prospectively evaluated the effects of dietary vitamin K intake on anticoagulation parameters. METHODS: In an observational protocol, clinical characteristics and vitamin K intake, assessed semiquantitatively, were evaluated in 39 outpatients who made 230 visits to our anticoagulation clinic. In a randomized crossover protocol, 12 patients with stable anticoagulation underwent 4-day in-hospital dietary interventions, 1 to 2 weeks apart, providing a 500% increase and an 80% decrease in vitamin K intake relative to the baseline level. RESULTS: Univariate analysis of the observational data demonstrated a progressive, statistically significant inverse association between the vitamin K intake score and different levels of anticoagulation. Multivariate logistic regression analysis showed that vitamin K intake was independently associated with both overcoagulation and undercoagulation. In the randomized protocol, the international normalized ratio increased from 2.6 +/- 0.5 at baseline to 3.3 +/- 0.9 at day 7 (P = 0.005) in subjects on the vitamin K-depleted diet and decreased from 3.1 +/- 0.8 at baseline to 2.8 +/- 0.6 at day 4 (P = 0.04) in those on the vitamin K-enriched diet. CONCLUSION: Our prospective data strengthen the concept that the interaction between vitamin K and coumarin is a clinically relevant, major independent factor that interferes with anticoagulation stability. PMID- 15121491 TI - Effects of ACE gene insertion/deletion polymorphism on response to spironolactone in patients with chronic heart failure. AB - BACKGROUND: Angiotensin-converting enzyme (ACE) is involved in the pathophysiology of chronic heart failure, and its activity is determined in part by a polymorphism of the ACE gene. We hypothesized that the benefits of spironolactone, which inhibits downstream elements of ACE-mediated abnormalities, may depend on ACE genotype. METHODS: We randomly assigned 93 chronic heart failure patients to treatment with spironolactone (n = 47) or to a control group (n = 46) and followed them for 12 months. Genotype for the insertion/deletion polymorphism of the ACE gene was determined by polymerase chain reaction. An echocardiographic examination was performed at baseline and at the end of the 12 months. RESULTS: The mean (+/- SD) age of the 93 patients was 62 +/- 9 years, and the mean New York Heart Association class was 2 +/- 1. The genotype was DD in 26 patients (28%). Forty-seven patients were assigned to spironolactone treatment (mean dose, 32 +/- 16 mg). In the treated group, only patients with a non-DD genotype showed significant improvement in left ventricular ejection fraction (3.0%; 95% confidence interval [CI]: 1.2% to 4.8%; P = 0.002), end-systolic volume (-23 mL; 95% CI: -36 to -11; P = 0.0005), and end-diastolic volume (-27 mL; 95% CI: -43 to -12; P = 0.001). In the multivariate analysis, the estimated net effect of treatment was 29 mL better (95% CI: -20 to 78 mL) for end-diastolic volume, 20 mL better (95% CI: -18 to 58 mL) for end-systolic volume, but 1.4% worse (95% CI: -3.4% to 6.2%) for left ventricular ejection fraction in patients with non-DD versus DD genotypes. CONCLUSION: The effects of spironolactone treatment on left ventricular systolic function and remodeling may in part depend on ACE genotype. PMID- 15121492 TI - The effect of pulmonary fibrosis on survival in patients with hypersensitivity pneumonitis. AB - PURPOSE: To determine the effect of pulmonary fibrosis on survival in an unselected group of patients with hypersensitivity pneumonitis. METHODS: We identified 72 patients with hypersensitivity pneumonitis confirmed by surgical lung biopsy in the database of the Clinical Interstitial Lung Disease Program at the National Jewish Medical and Research Center. All biopsy specimens were scored according to the presence or absence of fibrosis. Comparisons were made between patients with (fibrotic group) and without (nonfibrotic group) pathologic fibrosis. Vital status was ascertained and Kaplan-Meier curves were plotted. Cox regression analysis was used to determine predictors of survival. RESULTS: Forty six patients were classified as fibrotic and 26 as nonfibrotic. Twenty-nine percent had exposure to a bird antigen, 33% had exposure to a microbial antigen, and 38% had unknown exposure. Patients with fibrosis were significantly older, showed greater restrictive lung physiology, and had greater all-cause and respiratory mortality. Median survival in fibrotic patients was 7.1 years, which was significantly less than survival in those without fibrosis. In an age adjusted regression analysis, antigen class, symptom duration, and lung function had no effect on survival. Only the presence of pathologic fibrosis was predictive of increased mortality (hazard ratio = 6.01; 95% confidence interval: 1.68 to 21.45; P = 0.006). CONCLUSION: Pulmonary fibrosis is associated with diminished survival in patients with hypersensitivity pneumonitis. PMID- 15121493 TI - End-of-life care in a voluntary hospitalist model: effects on communication, processes of care, and patient symptoms. AB - PURPOSE: To assess the effects of hospitalist care on communication, care patterns, and outcomes of dying patients. METHODS: We examined the charts of 148 patients who had died at a community-based, urban teaching hospital, comparing the end-of-life care provided by community physicians and hospitalists. RESULTS: Patients of hospitalists and community-based physicians were similar in age, race, severity of acute illness, and difficulties with activities of daily living. After admission, hospitalists had discussions with patients or their families regarding care more often than did community physicians (91% [67/74] vs. 73% [54/74], P = 0.006) and were more likely to document these discussions themselves. Among patients who were "full code" at admission, there was a trend towards patients of hospitalists receiving comfort care more frequently at the time of death (50% [25/48] vs. 37% [15/40], P = 0.14). Although there were no differences in the use of medications such as long-acting opioids, no symptoms in the 48 hours prior to death were more likely to be noted for patients of hospitalists (47% [n = 35] vs. 31% [n = 23]), P = 0.03). After adjustment for confounding factors in multivariable models, only findings regarding documentation of discussions and symptoms remained statistically significant. CONCLUSION: Hospitalists at a community-based teaching hospital documented substantial efforts to communicate with dying patients and their families, which may have resulted in improved end-of-life care. PMID- 15121494 TI - Familial aggregation of hypertension treatment and control in the Genetic Epidemiology Network of Arteriopathy (GENOA) study. AB - PURPOSE: To assess if the treatment and control of hypertension aggregates in families. METHODS: The Genetic Epidemiology Network of Arteriopathy (GENOA) study enrolled sibships between 1997 and 1999, including 1329 hypertensive non-Hispanic blacks (1057 sibling pairs) from Jackson, Mississippi, 1133 hypertensive non Hispanic whites (859 sibling pairs) from Rochester, Minnesota, and 752 hypertensive Hispanic whites (627 sibling pairs) from Starr County, Texas. Hypertension awareness and drug treatment were ascertained at examination; control was defined by blood pressure levels <140/90 mm Hg. As a measure of familial aggregation, odds ratios were calculated to assess concordance between sibling pairs in the treatment and control of hypertension. RESULTS: Overall, 90.5% of subjects were aware of their hypertension; 90.6% of those who were aware were treated with antihypertensive drugs and 56.0% of those treated had their hypertension controlled. There was statistically significant sib-sib concordance in the treatment of hypertension (odds ratio [OR] = 1.61; 95% confidence interval [CI]: 1.25 to 2.47; P = 0.003) and in the control of drug-treated hypertension (OR = 1.51; 95% CI: 1.25 to 1.81; P <0.0001). CONCLUSION: These findings suggest that the treatment and control of hypertension aggregates in families. PMID- 15121495 TI - Exercise-based rehabilitation for patients with coronary heart disease: systematic review and meta-analysis of randomized controlled trials. AB - PURPOSE: To review the effectiveness of exercise-based cardiac rehabilitation in patients with coronary heart disease. METHODS: A systematic review and meta analysis of randomized controlled trials was undertaken. Databases such as MEDLINE, EMBASE, and the Cochrane Library were searched up to March 2003. Trials with 6 or more months of follow-up were included if they assessed the effects of exercise training alone or in combination with psychological or educational interventions. RESULTS: We included 48 trials with a total of 8940 patients. Compared with usual care, cardiac rehabilitation was associated with reduced all cause mortality (odds ratio [OR] = 0.80; 95% confidence interval [CI]: 0.68 to 0.93) and cardiac mortality (OR = 0.74; 95% CI: 0.61 to 0.96); greater reductions in total cholesterol level (weighted mean difference, -0.37 mmol/L [-14.3 mg/dL]; 95% CI: -0.63 to -0.11 mmol/L [-24.3 to -4.2 mg/dL]), triglyceride level (weighted mean difference, -0.23 mmol/L [-20.4 mg/dL]; 95% CI: -0.39 to -0.07 mmol/L [-34.5 to -6.2 mg/dL]), and systolic blood pressure (weighted mean difference, -3.2 mm Hg; 95% CI: -5.4 to -0.9 mm Hg); and lower rates of self reported smoking (OR = 0.64; 95% CI: 0.50 to 0.83). There were no significant differences in the rates of nonfatal myocardial infarction and revascularization, and changes in high- and low-density lipoprotein cholesterol levels and diastolic pressure. Health-related quality of life improved to similar levels with cardiac rehabilitation and usual care. The effect of cardiac rehabilitation on total mortality was independent of coronary heart disease diagnosis, type of cardiac rehabilitation, dose of exercise intervention, length of follow-up, trial quality, and trial publication date. CONCLUSION: This review confirms the benefits of exercise-based cardiac rehabilitation within the context of today's cardiovascular service provision. PMID- 15121496 TI - Exercise training for patients with heart failure: a systematic review of factors that improve mortality and morbidity. AB - PURPOSE: To determine the efficacy of exercise training and its effects on outcomes in patients with heart failure. METHODS: MEDLINE, Medscape, and the Cochrane Controlled Trials Registry were searched for trials of exercise training in heart failure patients. Data relating to training protocol, exercise capacity, and outcome measures were extracted and reviewed. RESULTS: A total of 81 studies were identified: 30 randomized controlled trials, five nonrandomized controlled trials, nine randomized crossover trials, and 37 longitudinal cohort studies. Exercise training was performed in 2387 patients. The average increment in peak oxygen consumption was 17% in 57 studies that measured oxygen consumption directly, 17% in 40 studies of aerobic training, 9% in three studies that only used strength training, 15% in 13 studies of combined aerobic and strength training, and 16% in the one study on inspiratory training. There were no reports of deaths that were directly related to exercise during more than 60,000 patient hours of exercise training. During the training and follow-up periods of the randomized controlled trials, there were 56 combined (deaths or adverse events) events in the exercise groups and 75 combined events in the control groups (odds ratio [OR] = 0.98; 95% confidence interval [CI]: 0.61 to 1.32; P = 0.60). During this same period, 26 exercising and 41 nonexercising subjects died (OR = 0.71; 95% CI: 0.37 to 1.02; P = 0.06). CONCLUSION: Exercise training is safe and effective in patients with heart failure. The risk of adverse events may be reduced, but further studies are required to determine whether there is any mortality benefit. PMID- 15121497 TI - Risks versus benefits of flexible sigmoidoscopy after myocardial infarction: an analysis of 78 patients at three medical centers. PMID- 15121498 TI - Vitamin K and oral anticoagulation: thought for food. PMID- 15121499 TI - From case report to meta-analysis--additional evidence for the benefits of exercise training in cardiac patients. PMID- 15121500 TI - Gastroesophageal reflux disease and diarrhea. PMID- 15121502 TI - A heart insulated by fat. PMID- 15121503 TI - The future of fellowship education in internal medicine. PMID- 15121504 TI - Storage of serum in plastic and glass containers may alter the serum concentration of polychlorinated biphenyls. AB - Valid exposure assessment and biomonitoring of toxicants rely on standardized specimen collection, handling, storage, and measurement. In a study designed to determine organochlorine concentrations in blood samples, we recruited participants from registered anglers in Michigan. After participants were interviewed, blood was collected from study subjects, either at home by a phlebotomist or in a commercial blood-draw station. The phlebotomists stored their samples in glass containers, but without our knowledge, the commercial laboratory transferred the specimens to plastic containers for freezing in its central facility. Samples were analyzed in the Analytical Chemistry Section Laboratory of the Michigan Department of Community Health. This laboratory also provided information on storage in glass (n = 28) versus plastic containers (n = 113). We conducted linear regression analyses to assess factors that may explain the concentrations of polychlorinated biphenyls (PCBs), dichlorodiphenyldichloroethylene (DDE), and polybrominated biphenyls (PBBs). Our results indicate that storage of serum in plastic containers altered the total concentrations of PCBs, in particular, the higher chlorinated PCBs (PCB-180 and PCB-199), but not DDE or PBBs. No other characteristics of the samples could explain the higher PCB values (0.75 micro g/L vs. 0.45 micro g/L; p = 0.025) of those stored in plastic containers. The proportion of PCB detects in both subsamples did not differ. Some preceding studies have provided information on whether specimens were stored in glass or plastic containers; however, a number of studies have not. We suggest the initiation of a new review process to determine whether these earlier reports were based on unbiased PCB determinations. We recommend standardizing specimen collection, handling, storage, and measurement, which is particularly necessary for newly emerging analytes. PMID- 15121505 TI - Chemical communication threatened by endocrine-disrupting chemicals. AB - Communication on a cellular level--defined as chemical signaling, sensing, and response--is an essential and universal component of all living organisms and the framework that unites all ecosystems. Evolutionarily conserved signaling "webs," existing both within an organism and between organisms, rely on efficient and accurate interpretation of chemical signals by receptors. Therefore, endocrine disrupting chemicals (EDCs), which have been shown to disrupt hormone signaling in laboratory animals and exposed wildlife, may have broader implications for disrupting signaling webs that have yet to be identified as possible targets. In this article, I explore common evolutionary themes of chemical signaling (e.g., estrogen signaling in vertebrates and phytoestrogen signaling from plants to symbiotic soil bacteria) and show that such signaling systems are targets of disruption by EDCs. Recent evolutionary phylogenetic data have shown that the estrogen receptor (ER) is the ancestral receptor from which all other steroid receptors have evolved. In addition to binding endogenous estrogens, ERs also bind phytoestrogens, an ability shared in common with nodulation D protein (NodD) receptors found in Rhizobium soil bacteria. Recent data have shown that many of the same synthetic and natural environmental chemicals that disrupt endocrine signaling in vertebrates also disrupt phytoestrogen-NodD receptor signaling in soil bacteria, which is necessary for nitrogen-fixing symbiosis. Bacteria-plant symbiosis is an unexpected target of EDCs, and other unexpected nontarget species may also be vulnerable to EDCs found in the environment. PMID- 15121506 TI - Retrospective time-trend study of polybrominated diphenyl ether and polybrominated and polychlorinated biphenyl levels in human serum from the United States. AB - Six polybrominated diphenyl ethers (PBDEs), one hexabromobiphenyl [polybrominated biphenyl (PBB)], and one hexachlorobiphenyl [polychlorinated biphenyl (PCB)] were measured in 40 human serum pools collected in the southeastern United States during 1985 through 2002 and in Seattle, Washington, for 1999 through 2002. The concentrations of most of the PBDEs, which are commercially used as flame retardants in common household and commercial applications, had significant positive correlations with time of sample collection, showing that the concentrations of these compounds are increasing in serum collected in the United States. In contrast, PCB and PBB levels were negatively correlated with sample collection year, indicating that the levels of these compounds have been decreasing since their phaseout in the 1970s. PMID- 15121507 TI - Synergistic interaction in simultaneous exposure to Streptomyces californicus and Stachybotrys chartarum. AB - The microbial exposure associated with health complaints in moldy houses consists of a heterogeneous group of components, including both living and dead bacteria, fungi, and their metabolites and active compounds. However, little is known about the interactions between different microbes and their metabolites, although the cytotoxicity and inflammatory potential of certain individual microbes have been reported. In this study, we investigated the inflammatory responses of mouse RAW264.7 macrophages after exposure to six indoor air microbes (Aspergillus versicolor, Penicillium spinulosum, Stachybotrys chartarum, Bacillus cereus, Mycobacterium terrae, and Pseudomonas fluorescens) alone and together with the actinomycete Streptomyces californicus. The production of nitric oxide, levels of the proinflammatory cytokines tumor necrosis factor alpha (TNF-alpha) and interleukin-6 (IL-6), and cytotoxicity were measured. The coexposure to Sta. chartarum and Str. californicus caused a synergistic increase in the production of IL-6 but not other cytokines. In further experiments, the metabolites from Sta. chartarum or from closely related fungi (atranones B and E, satratoxin G, trichodermin, 7-alpha-hydroxytrichodermol, staplabin, and SMTP-7) and the known fungal toxins sterigmatocystin, citrinin, and ochratoxin A were each tested with Str. californicus. The testing revealed a synergistic response in TNF-alpha and IL-6 production after coexposure to Str. californicus with both trichodermin and 7-alpha-hydroxytrichodermol. Finally, the synergistic inflammatory response caused by Str. californicus and trichodermin together was studied by analyzing for the presence of nuclear factor-kappa-B (NF-kappa-B) in nuclear extracts of the exposed cells. The exposure to Str. californicus induced the binding of NF kappa-B proteins to the NF-kappa-B consensus sequence as well as to the natural NF-kappa-B site of the IL-6 promoter. Adding trichodermin to the exposure did not increase the DNA binding. PMID- 15121508 TI - Urinary 8-hydroxy-2'-deoxyguanosine as a biomarker of oxidative DNA damage in workers exposed to fine particulates. AB - Residual oil fly ash (ROFA) is a chemically complex mixture of compounds, including metals that are potentially carcinogenic because of their ability to cause oxidative injury. In this study, we investigated the association between exposure to particulate matter with an aerodynamic mass median diameter 4,000 school-aged children in 12 southern California communities. Cases were defined as physician-diagnosed asthma by age 5, and controls were asthma-free at study entry, frequency-matched on age, sex, and community of residence and countermatched on in utero exposure to maternal smoking. Telephone interviews were conducted with mothers to collect additional exposure and asthma histories. Conditional logistic regression models were fitted to estimate odds ratios (ORs) and 95% confidence intervals (CIs). Asthma diagnosis before 5 years of age was associated with exposures in the first year of life to wood or oil smoke, soot, or exhaust (OR = 1.74; 95% CI, 1.02-2.96), cockroaches (OR = 2.03; 95% CI, 1.03-4.02), herbicides (OR = 4.58; 95% CI, 1.36 15.43), pesticides (OR = 2.39; 95% CI, 1.17-4.89), and farm crops, farm dust, or farm animals (OR = 1.88; 95% CI, 1.07-3.28). The ORs for herbicide, pesticide, farm animal, and crops were largest among children with early-onset persistent asthma. The risk of asthma decreased with an increasing number of siblings (ptrend = 0.01). Day care attendance within the first 4 months of life was positively associated with early-onset transient wheezing (OR = 2.42; 95% CI, 1.28-4.59). In conclusion, environmental exposures during the first year of life are associated with childhood asthma risk. PMID- 15121524 TI - Chemical process safety at a crossroads. PMID- 15121523 TI - Probability estimates for the unique childhood leukemia cluster in Fallon, Nevada, and risks near other U.S. Military aviation facilities. AB - A unique cluster of childhood leukemia has recently occurred around the city of Fallon in Churchill County, Nevada. From 1999 to 2001, 11 cases were diagnosed in this county of 23,982 people. Exposures related to a nearby naval air station such as jet fuel or an infectious agent carried by naval aviators have been hypothesized as potential causes. The possibility that the cluster could be attributed to chance was also considered. We used data from the Surveillance, Epidemiology, and End Results Program (SEER) to examine the likelihood that chance could explain this cluster. We also used SEER and California Cancer Registry data to evaluate rates of childhood leukemia in other U.S. counties with military aviation facilities. The age-standardized rate ratio (RR) in Churchill County was 12.0 [95% confidence interval (CI), 6.0-21.4; p = 4.3 times symbol 10( 9)]. A cluster of this magnitude would be expected to occur in the United States by chance about once every 22,000 years. The age-standardized RR for the five cases diagnosed after the cluster was first reported was 11.2 (95% CI, 3.6-26.3). In contrast, the incidence rate was not increased in all other U.S. counties with military aviation bases (RR = 1.04; 95% CI, 0.97-1.12) or in the subset of rural counties with military aviation bases (RR = 0.72; 95% CI, 0.48-1.08). These findings suggest that the Churchill County cluster was unlikely due to chance, but no general increase in childhood leukemia was found in other U.S. counties with military aviation bases. PMID- 15121525 TI - Hazards of fast food. PMID- 15121526 TI - Asbestos and international organizations. PMID- 15121527 TI - Exceeding the methyl mercury reference dose: how dangerous is it? PMID- 15121529 TI - Arsenic in food. PMID- 15121530 TI - Food and population growth. PMID- 15121531 TI - Human testing: Sass and Needleman respond to industry. PMID- 15121532 TI - Origins of obesity. PMID- 15121533 TI - Awards spur excellence in research. PMID- 15121534 TI - Lessons learned? Chemical plant safety since Bhopal. PMID- 15121535 TI - Environmental buyouts: protection at a price. PMID- 15121536 TI - Keeping an eye on exposure: video monitoring in the lab. PMID- 15121538 TI - A meta-analysis of priming effects on impression formation supporting a general model of informational biases. AB - Priming researchers have long investigated how providing information about traits in one context can influence the impressions people form of social targets in another. The literature has demonstrated that this can have 3 different effects: Sometimes primes become incorporated in the impression of the target (assimilation), sometimes they are used as standards of comparison (anchoring), and sometimes they cause people to consciously alter their judgments (correction). In this article, we present meta-analyses of these 3 effects. The mean effect size was significant in each case, such that assimilation resulted in impressions biased toward the primes, whereas anchoring and correction resulted in impressions biased away from the primes. Additionally, moderator analyses uncovered a number of variables that influence the strength of these effects, such as applicability, processing capacity, and the type of response measure. Based on these results, we propose a general model of how irrelevant information can bias judgments, detailing when and why assimilation and contrast effects result from default and corrective processes. PMID- 15121539 TI - A recurrent connectionist model of person impression formation. AB - Major findings in impression formation are reviewed and modeled from a connectionist perspective. The findings are in the areas of primacy and recency in impression formation, asymmetric diagnosticity of ability- and morality related traits, increased recall for trait-inconsistent information, assimilation and contrast in priming, and discounting of trait inferences by situational information. The majority of these phenomena are illustrated with well-known experiments and simulated with an autoassociative network architecture with linear activation update and using the delta learning algorithm for adjusting the connection weights. All of the simulations successfully reproduced the empirical findings. Moreover, the proposed model is shown to be consistent with earlier algebraic models of impression formation (Anderson, 1981; Busemeyer, 1991; Hogarth and Einhorn, 1992). The discussion centers on how our model compares to other connectionist approaches to impression formation and how it may contribute to a more parsimonious and unified theory of person perception. PMID- 15121540 TI - Culture and aggression-from context to coercion. AB - This article is an attempt to study the neglected linkages between culture and aggression. It does so by conceptualizing culture as a set of affordances and constraints that channel the expression of coercive means of social control by self and others. All cultural systems represent solutions to the problems associated with distributing desired material and social resources among its group members while maintaining social order and harmony. Norms are developed surrounding the exercise of mutual influence in the process of resource allocation, favoring some and marginalizing others. Violations of these norms by resource competitors are conceptualized as "aggressive" behaviors and stimulate a process of justified counter attack, escalating the violence. The current data from both societal-level and individual-level studies are examined and integrated in light of this organizing framework, and future studies are proposed to explore the interface between culture and aggression more productively. PMID- 15121541 TI - Social consequences of disparagement humor: a prejudiced norm theory. AB - In this article we introduce a "prejudiced norm theory" that specifies the social psychological processes by which exposure to disparagement humor uniquely affects tolerance of discrimination against members of groups targeted by the humor. Our theory posits that a norm of tolerance of discrimination implied by disparagement humor functions as a source of self-regulation for people high in prejudice. For people high in prejudice, this norm regulates the effect of exposure to disparagement humor on tolerance of subsequently encountered discriminatory events. Our theory contributes to the literature on prejudice and discrimination by delineating the processes by which disparagement humor creates a normative climate of tolerance of discrimination, as well as variables that accentuate and attenuate its effects. PMID- 15121543 TI - Mechanical influences on cells, tissues and organs - 'Mechanical Morphogenesis'. AB - Cells can sense changes in their mechanical environment and promote alterations and adaptations in tissue structure and function. Mechanical stimuli regulate such fundamental processes as cell division and differentiation and determine tissue form. The current editorial outlines the general scope of a subject area we have called 'mechanical morphogenesis'. We are promoting it as an area of special interest for future issues of the European Journal of Morphology. Clearly, mechanical loading is of pivotal importance to the development, function and repair of all tissues in the musculoskeletal system, including bone, ligament, tendon, skeletal muscle, intervertebral disc and meniscus. Bone in particular has attracted special interest and mechanical strain is central to both Wolff 's law and Frost's 'mechanostat' model of bone behaviour. But it is skeletal muscle that shows the most obvious and rapid response to altered load, with striated muscle fibres hypertrophing with strength-training programmes, and atrophing in the absence of adequate mechanical stimulation. Articular cartilage, together with tendons and ligaments is also responsive to changing exercise levels, and either abnormally high or low loads are detrimental. However, the influence of mechanical forces extends to many other organ systems, including the respiratory, cardiovascular, nervous and integumentary systems. The bronchial mucosa and the alveoli are subject to tensile and compressive loading during the volume changes that occur in respiration, and surface tension is also of paramount importance. The whole form of the cardiovascular system is driven by the haemodynamic influences of blood, and atherosclerosis has an underlying mechanical basis. The characteristic plaques tend to occur at sites of obvious mechanical significance - regions of arterial branching and curvature, where shear stress on the vessel wall may be low, but tensile stress high. Sensory perception by the nervous system has a well known mechanical basis and the cochlea is perhaps the most elaborate example of a site where sensory cells transduce mechanical forces and relay information to the brain. Mechanical force has also been proposed as a regulating factor in controlling axonal growth. Finally, the integumentary system has several structural adaptations that obviously relate to the influence of mechanical forces. The thickened layer of keratinised squames in the palms and soles is directly related to the high levels of shear at these locations. PMID- 15121544 TI - The epithelia of the protrusible tongue of Eurycea longicauda guttolineata (Hoolbrook 1838) (Urodela: Plethodontidae). AB - In this study the lingual and sublingual glands, the lingual stem and the epithelial surface of the protrusible secondary tongue were investigated by light, scanning and transmission electron microscopy. The quality of the secretions of the epithelia was characterized histochemically. The lingual epithelium is formed by superficial (pavement) and goblet cells and at the margin of the tongue pad are also regions covered by ciliated cells. On the dorsal part of the tongue there are goblet cells of type A with mainly acidic secretions and of type B containing neutral secretions. Most of the goblet cells on the ventral side of the tongue (hypoglottis) show a strong alcian blue/PAS positive reaction (type I) and some produce neutral secretions (type II). The glandular cells of the lingual gland react positively to alcian blue and PAS in the apical region of the gland. In contrast there is only alcian blue-positive staining in the basal part of the gland. The size and complexity of the inclusion bodies of the secretory granules increase in a basal direction. In addition, there are ciliated cells in the glandular epithelium. Although the epithelium of the lingual stem is thin, it is double-layered. The cell types observed in this region are identical to those of the ventral part of the protrusible tongue. At the margin of the sublingual gland are trough-like structures. In the center, tubular parts are observed. The cells of this gland are stain strongly with alcian blue (pH 1.0) mainly in the basal part of the gland. The results of this are compared to the tongue pad and the lingual gland of Salamandra salamandra and Ambystoma mexicanum. PMID- 15121545 TI - Tissue collection methods for antler research. AB - The rapid growth of deer antlers makes them potentially excellent models for studying tissue regeneration. In order to facilitate this, we have developed and refined antler tissue sampling methods through years of antler research. In the study, antler tissues were divided into three main groups: antler stem tissue, antler blastema and antler growth centre. For sampling stem tissue, entire initial antlerogenic periosteum (around 22 mm in diameter) could be readily peeled off from the underlying bone using a pair of rat-toothed forceps after delineating the boundary. Apical and peripheral periosteum/ perichondrium of pedicle and antler could only be peeled off intact when they were cut into 4 quadrants and 0.5 cm-wide strips respectively. Antler blastema included blastema per se, and potentiated and dormant periostea. Blastema per se was sampled after it was divided into 4 quadrants using a disposable microtome blade. Potentiated and dormant periostea were collected following the same method used for sampling peripheral periosteum of pedicle and antler. The antler growth centre was divided with a scalpel into 5 layers according to distinctive morphological markers. The apical skin layer could be further separated into dermis and epidermis using enzyme digestion for the study of tissue interaction. We believe that the application of modern techniques coupled with the tissue collection methods reported here will greatly facilitate the establishment of these valuable models. PMID- 15121546 TI - Anatomical variations of the cystic artery. AB - Thorough knowledge about the origin of the cystic artery is surgically important, especially when intraoperative or post-operative bleeding occurs in the gallbladder fossa. The arterial supply of the gallbladder was studied in 81 livers. The gallbladder was supplied by one cystic artery in 86% and by two arteries in 14% of cases. When a single artery was present, it originated from the right hepatic artery in 53% of livers. Other origins included the anterior or the posterior sectional hepatic artery, the replacing right hepatic artery, and in 5% of cases, segmental arteries for segments 4, 5, 6 and 8. When two cystic arteries supplied the gallbladder, both most commonly originated from the right hepatic artery (7% incidence). In 1% of cases, a subsegmental branch for segment 6 and a subsegmental branch for segment 5 respectively, originated from the cystic artery. PMID- 15121547 TI - Morphological and functional changes in cell junctions during secretory stimulation in the perfused rat submandibular gland. AB - To examine the influence of cholinergic and beta-adrenergic agents on paracellular transport, we applied confocal microscopy and freeze-fracture to the isolated, perfused submandibular gland of the rat. By confocal microscopy, perfusion of lucifer yellow through an arterial catheter, revealed a bright fluorescence in the basolateral spaces of acini, but not in the intercellular canaliculi. However, addition of isoproterenol on carbachol stimulation, induced lucifer yellow fluorescence in intercellular canaliculi. This finding indicates that isoproterenol is capable of opening the paracellular route. The tight junction strands surrounding intercellular canaliculi were visualized using freeze replicas. Fixation was carried out both by vascular perfusion with Karnovsky's solution and by metal contact rapid freezing with liquid helium. In the chemically-fixed specimens, the strand particles of tight junctions formed 2 5 lines at the P-face along most of the apical portion at rest. With carbachol/isoproterenol stimulation, the strand particles rearranged with free ends and terminal loops. In the rapidly frozen specimens, the strand particles were arranged more irregularly even in the resting state. The meshwork of strands became more disheveled and interrupted during carbachol/ isoproterenol stimulation. The present findings led us to conclude that: 1) the beta-adrenergic agent, isoproterenol, can open the paracellular transport. 2) in the rapidly frozen specimen, the tight junction strand particles are arranged roughly and become disheveled and interrupted during stimulation by carbachol/isoproterenol. These findings may be related to rearrangement of subcellular structures, especially of the actin filament network. PMID- 15121548 TI - A rare variation in the high division of the sciatic nerve surrounding the superior gemellus muscle. AB - The sciatic nerve normally leaves the pelvis by passing through the greater sciatic foramen below piriformis. However, it may divide into its common fibular and tibial nerve components within the pelvis and its relationship with piriformis is variable. In this paper, we describe a new anatomical variation in which the common fibular nerve passed superior, and the tibial nerve inferior, to the superior gemellus muscle. Anatomical variations such as these may contribute to piriformis syndrome, coccygodynia and muscle atrophy. PMID- 15121549 TI - Histochemistry of glycoconjugates in the nasolabial skin of the Japanese serow (Capricornis crispus, Artiodactyla, Mammalia) with special reference to glandular structures. AB - The histochemistry of glycoconjugates in the nasolabial skin of the Japanese serow ( Capricornis crispus ) was studied by light microscopic histochemical methods, particularly lectin histochemistry. The eccrine glands present exhibited neutral and acidic glycoconjugates with different saccharide residues (alpha-L fucose, beta-D-galactose, beta-N-acetyl-D-glucosamine, alpha-D-galactose and N acetyl-neuraminic acid) especially in the cells of the secretory acini, the free surface of the collecting duct cells also showed distinct positive reactions with most of the histochemical methods. The thick epidermis of the nasolabial skin contained smaller amounts of glycoproteins. The results obtained are discussed with regard to possible functions of the glandular secretions. This substance mixture may particularly improve water retention on the skin surface, and protect against physical damage as well as microbial contamination. There seem to be no basic differences of muzzle functions between wild and domesticated bovine species. PMID- 15121552 TI - Hysterectomy versus expanded medical treatment for abnormal uterine bleeding: clinical outcomes in the medicine or surgery trial. AB - OBJECTIVE: To compare clinical outcomes after randomization to hysterectomy versus medical treatment in patients with chronic abnormal uterine bleeding refractory to medroxyprogesterone acetate. METHODS: We randomly assigned 63 premenopausal women with abnormal uterine bleeding refractory to cyclic medroxyprogesterone acetate treatment to receive either a hysterectomy or expanded medical treatment. Within each randomized group, the specific treatment approach was determined by patient and provider preference. The primary analysis compared changes in clinical outcomes at 6 and 24 months by using an intention-to treat approach. Secondary as-treated analyses after adjustment for baseline covariates compared participants randomly assigned to medical treatment who continued the medical approach with those who crossed over to hysterectomy. RESULTS: The intention-to-treat analyses at 6 months revealed greater symptom improvement in the hysterectomy group than in the medicine group for pelvic pain (P <.01), urinary urgency (P =.03), incomplete bladder emptying (P =.03), breast pain (P =.02), and cessation of vaginal bleeding (87% versus 11%, P <.001). Seventeen of 32 women assigned to medicine (53%) eventually crossed over and received a hysterectomy, and by 24 months the statistically significant differences by intention-to-treat were limited to greater improvement in hot flushes (P <.01) and cessation of vaginal bleeding (P <.01). Within-group analyses at year 2 showed statistically significant improvements from baseline on most symptoms for women who had a hysterectomy, whether through randomization or crossover. Women remaining on medical treatments had statistically significant improvements in pelvic pain, pelvic/bladder pressure, and stress incontinence. In a nonrandomized comparison with women who remained on medical treatments through year 2, those crossing over to hysterectomy experienced greater improvements in bleeding (P <.01), pelvic pain (P <.01), low back pain (P =.02), breast pain (P =.01), urinary frequency (P =.01), and urgency (P =.02). However, they also experienced more days off from work or usual activities (P <.01) and more days spent in bed (P <.01) than those who remained on medicine. CONCLUSION: For patients with abnormal uterine bleeding refractory to medroxyprogesterone acetate, hysterectomy is superior to expanded efforts with oral medications for alleviating clinical symptoms but may lead to more days of restricted activity. PMID- 15121553 TI - Resource use for total and supracervical hysterectomies: results of a randomized trial. AB - OBJECTIVE: Hysterectomy is the most common major surgical procedure performed in the United States for nonobstetric reasons. Although most hysterectomies include removal of the cervix, the rate of supracervical procedures has increased in recent years. To provide evidence about the outcomes of both types of hysterectomy, we conducted a randomized clinical trial of total (TAH) or supracervical (SCH) hysterectomy (the "TOSH" trial). We report here an analysis of 24-month resource use by patients in this trial. METHODS: A randomized controlled trial was performed at 3 clinical centers to compare resources used by 120 patients who received a total or supracervical abdominal hysterectomy. Service use during a 24-month follow-up period was identified from medical and billing records and patient reports. Each service used was assigned a relative value, which was then converted into 2002 U.S. dollars. RESULTS: Overall resource use was similar in the 2 study groups in the first 12 months after randomization (TAH 5,870 US dollars; SCH 6,018 US dollars; 95% confidence interval for difference -960 US dollars, 1,255 US dollars; P <.79) and for the full 24 months (TAH 6,448 US dollars; SCH 7,479 US dollars; 95% confidence interval for difference -533 US dollars, 2,616 US dollars; P <.20). In exploratory multivariable analyses, resource use was significantly associated with baseline body mass index greater than or equal to 35 kg/m(2) (8,440 US dollars versus 6,398 US dollars, P =.02) and heavy bleeding (7,550 US dollars versus 5,368 US dollars, P =.02). CONCLUSION: We conclude that the use of medical care resources over a 24-month period is comparable for total and supracervical hysterectomy. The association of a woman's weight and bleeding pattern with subsequent resource use requires further investigation. PMID- 15121554 TI - Periurethral masses: etiology and diagnosis in a large series of women. AB - OBJECTIVE: To describe the differential diagnosis of periurethral masses in a consecutive series extracted from a single tertiary urogynecologic practice database. METHODS: A patient database of a private urology and urogynecology practice with 1,950 women was searched for patients who were found to have a periurethral mass during the accrual dates of 1994 to 2002, and these records were reviewed for diagnostic testing and results. All women provided a history, completed a questionnaire, and underwent physical examination, voiding diary, cystoscopy, and videourodynamic testing; selected patients then underwent additional imaging. RESULTS: Seventy-nine (4%) patients aged 41.2 +/- 14 years were identified. Of these, 72 (91%) had been referred for evaluation of persistent irritative lower urinary tract symptoms or incontinence. Seven patients (9%) had been referred specifically because of a periurethral mass. Sixty-six patients (84%; 95% confidence interval [CI] 73%, 91%) had urethral diverticula, of which 4 (6%; 95% CI 2%, 14.8%) contained malignancies. Six patients (7%; 95% CI 3%, 15%) had vaginal cysts histologically identified as fibromuscular tissue, 4 (5%; 95% CI 1%, 12%) had leiomyomata, and 2 (2.5%; 95% CI 0.03%, 8.8%) had ectopic ureteroceles. Two patients had vaginal squamous cell carcinomas (2.5%; 95% CI 0.03%, 8.8%), and 1 had an infected granuloma. Masses were palpable in 42 patients (53.8%; 95% CI 42%, 64%) and in 37 patients either were encountered at surgery (n = 5) or were urethral diverticula diagnosed by voiding cystourethrogram (n = 32). CONCLUSION: Periurethral masses were encountered in less than 4% of our patient sample. Most masses were urethral diverticula; however, the differential diagnosis included leiomyoma, vaginal cysts, and malignancy. Masses were generally either palpable or seen at imaging studies performed during evaluation of lower urinary tract symptoms. PMID- 15121555 TI - The risk of pregnancy after vasectomy. AB - OBJECTIVE: To describe the pregnancy rates among women whose husbands underwent vasectomy. METHODS: Between 1985 and 1987, 573 women aged 18-44 years whose husbands underwent vasectomy in medical centers in 5 U.S. cities were enrolled in the U.S. Collaborative Review of Sterilization, a prospective cohort study of male and female sterilization. Women were interviewed by telephone at 1, 2, 3, and 5 years after their husbands underwent vasectomy. RESULTS: Among the 540 eligible women at risk for pregnancy, there were 6 pregnancies occurring from 6 to 72 weeks after vasectomy. The cumulative probability of failure per 1,000 procedures (95% confidence interval) was 7.4 (0.2, 14.6) 1 year after vasectomy and 11.3 (2.3, 20.3) at years 2, 3, and 5 [corrected]. CONCLUSION: Couples considering vasectomy should be counseled about the small, but real, risk of pregnancy following the procedure and that men are not sterile immediately after vasectomy. PMID- 15121556 TI - A randomized comparison of misoprostol 6 to 8 hours versus 24 hours after mifepristone for abortion. AB - OBJECTIVE: To demonstrate equivalence between mifepristone 200 mg followed 6 to 8 hours later and 24 hours later by misoprostol 800 microg vaginally for abortion in women up to 63 days of gestation. METHODS: Mifepristone 200 mg was swallowed by 1,080 women after which they were randomly assigned to self-administer misoprostol intravaginally 6 to 8 hours later (group 1) or 23 to 25 hours later (group 2) at home. Participants returned for an evaluation, including transvaginal ultrasonography, 7 +/- 1 days after initiating treatment. Subjects who had not aborted were offered a second dose of misoprostol. All participants returned approximately 2 weeks after receiving mifepristone. Telephone contact was also attempted approximately 5 weeks after treatment. Treatment was considered a failure if a suction aspiration was performed for any indication. RESULTS: Complete abortion rates for groups 1 and 2 were 503 of 525 (95.8%, 95% confidence interval 93.7%, 97.3%) and 521 of 531 (98.1%, 95% confidence interval 96.6%, 99.1%), respectively, which were statistically equivalent. Side effects were significantly more common after mifepristone administration for women in group 2. Nausea, vomiting, and heavy bleeding were also significantly greater for women in group 2 after misoprostol treatment. Pain and subject acceptability were similar between groups. CONCLUSION: Mifepristone 200 mg followed 6 to 8 hours later by misoprostol 800 microg vaginally is as effective for abortion and has significantly fewer side effects as compared with regimens using a 24-hour dosing interval. Women receiving mifepristone and vaginal misoprostol for abortion can have the flexibility to administer the misoprostol as soon as 6 hours after using the mifepristone. PMID- 15121557 TI - Two regimens of misoprostol for treatment of incomplete abortion. AB - OBJECTIVES: Misoprostol shows promise for treatment of incomplete abortion. We evaluated 2 simple misoprostol regimens to estimate whether they were effective in treating incomplete abortion. METHODS: A total of 169 women was randomly assigned to either a single or double dose of 600 microg misoprostol. The women, who would have received a surgical evacuation of the uterus for incomplete abortion, were patients at 2 hospitals in Bangkok, Thailand. The 2 groups of women were compared for success of treatment (no need for surgical evacuation), side effects, and acceptability. RESULTS: Sixty-six percent of women in the single-dose group and 70% of women in the double-dose group had complete abortions with misoprostol. More than 90% of women in the single- and double-dose groups reported that the side effects were tolerable; frequency of side effects was similar between the 2 groups. Women found the treatment acceptable. Approximately 90% of women in both groups would recommend the treatment to a friend. Acceptability and efficacy were different at the 2 participating clinics. CONCLUSION: Misoprostol is an effective treatment for incomplete abortion. Simple regimens may be as effective as more complicated ones and a single dose of 600 microg should be further evaluated in larger trials. PMID- 15121558 TI - Oral, rectal, and vaginal pharmacokinetics of misoprostol. AB - OBJECTIVE: To compare the pharmacokinetic profiles of orally, rectally, and vaginally administered misoprostol tablets in pregnant women. METHODS: Women between 7 and 14 completed weeks of gestation were recruited and randomly assigned to be given 400 microg misoprostol orally, rectally, or vaginally 3 hours before surgical termination of pregnancy. Blood samples were obtained at 0, 7.5, 15, 30, 45, 60, 90, 120, and 240 minutes and later analyzed for plasma concentrations of misoprostol free acid (the principle metabolite). RESULTS: Vaginal misoprostol was present in the circulation longer than oral misoprostol and had a greater area under curve at 240 minutes (P <.001). Rectal misoprostol had a similar pattern but a much lower area under curve at 240 minutes. Oral misoprostol had a significantly greater peak plasma concentration and a shorter duration to maximum concentration than either rectal or vaginal misoprostol (both P <.001). CONCLUSION: Oral misoprostol tablet is also absorbed by the rectal and vaginal routes. Misoprostol administered in early pregnancy has route-dependent pharmacokinetics and is absorbed best when administered vaginally. LEVEL OF EVIDENCE: I PMID- 15121559 TI - Ultrasound evaluation of the endometrium after medical termination of pregnancy. AB - OBJECTIVE: To determine ultrasound parameters associated with the need for clinical intervention after mifepristone and misoprostol termination of pregnancy. METHODS: Charts of patients undergoing medical termination according to a standard protocol in a 13-month period were reviewed. Endometrial thickness and the presence of gestational sac, fluid interface, or complex echoes on postprocedure ultrasonogram were recorded. Repeat doses of medication, surgical intervention, and complications were noted. Success was defined as an abortion completed after a single course of medical therapy. RESULTS: Postprocedure ultrasonograms were available for 525 of 684 patients. Endometrial thickness was measurable in 437 cases. The observed mean endometrial thickness was 4.10 +/- 1.80 mm (range 0.67-13.4 mm). Endometrial thickness was inversely proportional to the number of days after initiation of therapy when ultrasonography was performed (r = -0.22; P <.001). The endometrium was thicker in the women who had failed than in those who had a successful medical abortion (6.15 +/- 1.95 mm [range 3.35 10.0 mm] versus 4.01 +/- 1.75 mm [range 0.67-13.4 mm], respectively; P <.001), but the wide overlap in endometrial thicknesses nullified the clinical usefulness of this difference. CONCLUSION: Endometrial thickness after administration of a single dose of mifepristone and misoprostol for medical termination should not dictate clinical intervention. The decision to treat should be based on the presence of a persistent gestational sac or compelling clinical signs and symptoms. PMID- 15121560 TI - Increased plasma carnitine concentrations in preeclampsia. AB - OBJECTIVE: Preeclampsia is associated with abnormal lipid metabolism, including fatty acid metabolism. Carnitine plays an indispensable role in the oxidation of fatty acids. The aim of the study was to evaluate the possible role of abnormal fatty acid oxidation in preeclampsia by comparing plasma carnitine levels between preeclamptic and healthy control pregnant women. METHODS: Plasma concentrations of free carnitine and short-, medium-, and long-chain acylcarnitines were investigated with electrospray tandem mass spectrometry in pregnant women with preeclampsia (n = 33) and in normotensive healthy pregnant control subjects (n = 28). Excluded were multiple pregnancies and women with preexistent hypertension, diabetes, renal dysfunction, immune disease, and intrauterine fetal death. Control subjects were healthy pregnant women without hypertension or proteinuria. RESULTS: The results revealed that, except for the medium-chain plasma acylcarnitines, all plasma carnitines were significantly increased (P <.001) in the preeclamptic group (t test for unpaired samples). Free carnitine and the short- and long-chain acylcarnitine values were increased by approximately 50% compared with the control group. Total and short-chain plasma acylcarnitine levels were significantly correlated to diastolic blood pressure, whereas no relationship could be demonstrated between carnitine concentrations and the variables proteinuria and systolic blood pressure. CONCLUSION: The considerable increased plasma carnitine concentrations, together with the accumulation of lipids, support the role of abnormal lipid metabolism in the pathophysiology of preeclampsia. It is suggested that toxic metabolites resulting from abnormal fatty acid oxidation in the placenta contribute to the endothelial dysfunction of preeclampsia. PMID- 15121561 TI - Uterine effects of estrogen plus progestin therapy and raloxifene: adjudicated results from the EURALOX study. AB - OBJECTIVE: To compare the incident rate of abnormal endometrial findings in postmenopausal women receiving treatment with either 60 mg of raloxifene or a continuous combined estrogen plus progestin therapy containing 2 mg of 17 beta estradiol plus 1 mg of norethisterone acetate for a duration of up to 12 months. METHODS: One thousand eight asymptomatic postmenopausal women with osteoporosis or cardiovascular risk factors with an endometrial thickness of less than 5 mm at baseline participated in this prospective, randomized, double-blind trial that lasted 6 months; 347 of these women also participated in a 6-month extension. Women with repeated bleeding or an increase in endometrial thickness to above 5 mm were subjected to saline-infused sonohysterography or hysteroscopy with biopsy. Sonographic, histologic, and clinical findings were adjudicated by a panel of 4 experts blinded with respect to patients' treatments. All adjudicated patients were grouped into 15 diagnostic categories according to predefined criteria. RESULTS: Three hundred thirty-four women needed adjudication during the core phase, 73 (14.7%) of those taking raloxifene and 261 (50.9%) taking continuous combined estrogen plus progestin therapy (P <.001). Compared with raloxifene, women using continuous combined estrogen plus progestin therapy had significantly higher rates of benign endometrial proliferation (8.8 versus 1.2%, P <.001), endometrial polyps (4.3 versus 2.0%, P =.048), and cystic atrophy (5.5 versus 1.2%, P <.001). CONCLUSION: Women using continuous combined estrogen plus progestin therapy more often have benign endometrial pathology and, in our study, more often required the protocol-specific gynecological follow-up assessments for safety reasons, as compared with those using raloxifene. These findings are of clinical relevance when choosing the most appropriate therapy for postmenopausal health risks such as osteoporosis. PMID- 15121562 TI - Distribution of soy-derived phytoestrogens in human breast tissue and biological fluids. AB - OBJECTIVE: Soy-derived phytoestrogens may exert several health-beneficial effects. Although plasma and urine levels of these compounds after ingestion have been thoroughly investigated, little is known about their tissue distribution, which is particularly important for tissues with high endogenous estrogen and estrogen receptor concentrations. We aimed to investigate the concentrations of genistein, daidzein, and equol in human breast tissue homogenate and to compare these with the corresponding values in serum and urine. METHODS: A randomized, double-blind, placebo-controlled study was undertaken to evaluate the concentrations of soy-derived phytoestrogens achieved in breast tissue homogenate, serum, and urine after ingestion of either a soy-based food supplement (n = 9) or a placebo tablet (n = 19) for 5 consecutive evenings before aesthetic breast surgery. To account for the heterogeneity of the breast tissue samples, markers for cellularity, epithelial content, blood vessel content, and total fat were determined. RESULTS: Urine concentrations of genistein, daidzein, and equol were significantly higher in the soy-supplemented subjects than in the subjects ingesting the placebo (P <.05). Only genistein was found to be significantly higher in serum of the soy group than in the placebo group, and no significant differences were found in breast tissue homogenate concentrations of all analytes between the 2 groups. CONCLUSION: Intake of soy-based food supplements for 5 consecutive days did not result in significantly higher genistein, daidzein, and equol concentrations in breast tissue homogenate when compared with the placebo group. The concentrations were in the low nanomolar range, whereas in the corresponding serum samples, concentrations were a hundred fold higher. PMID- 15121563 TI - Effects of hormonal contraception on bone mineral density after 24 months of use. AB - OBJECTIVE: To measure the effect of 24 months of depot medroxyprogesterone acetate use on bone mineral density compared with oral contraception (pills) and nonhormonal contraception. METHODS: Women aged 18-33 years self-selected oral contraception, depot medroxyprogesterone acetate, or nonhormonal contraception (controls). Those selecting pills were randomized to formulations containing either 35 microg ethinyl estradiol and norethindrone or 30 microg ethinyl estradiol and desogestrel. Controls were frequency matched on age and race/ethnicity to hormonal contraception users. Dual-energy X-ray absorptiometry of the lumbar spine (L1-L4) was performed at baseline, 12 months, and 24 months. Percent change in bone mineral density was analyzed by using analysis of covariance, adjusting for age, race/ethnicity, weight-bearing exercise, calcium intake, smoking status, and body mass index. RESULTS: Of the 191 women making up the final sample, 86 used pills, 47 used depot medroxyprogesterone acetate, and 58 used nonhormonal contraception. Women using depot medroxyprogesterone acetate for 24 months experienced, on average, a 5.7% loss in bone mineral density, with a 3.2% loss occurring between months 12 and 24. On average, users of desogestrel pills experienced a 2.6% loss in bone mineral density after 24 months. Bonferroni adjusted pairwise comparisons demonstrated that bone mineral density changes from baseline to 24 months among depot medroxyprogesterone acetate users differed significantly from changes experienced by either of the pill groups or the control group. Changes in bone mineral density among users of either pill did not significantly differ from each other or from controls. CONCLUSION: Loss of bone mineral density associated with depot medroxyprogesterone acetate use appears to be linear during the first 2 years of use. Shifts in bone mineral density among pill users were not significant when compared with controls. PMID- 15121564 TI - Maternal morbidity associated with vaginal versus cesarean delivery. AB - OBJECTIVE: To describe postpartum maternal morbidity associated with mode of delivery in term, singleton pregnancies. METHODS: The Magee Obstetric Medical and Infant database was examined for the years 1995 to 2000. Patients were grouped into 6 types of delivery mode: spontaneous vaginal delivery, operative vaginal delivery, primary cesarean delivery without trial of labor, primary cesarean delivery with trial of labor, repeat cesarean delivery without trial of labor, and repeat cesarean delivery with trial of labor. Multivariable logistic regression provided odds ratios and 95% confidence intervals (CI) for morbidity by delivery mode adjusted for demographic characteristics and comorbidities. Spontaneous vaginal delivery was used as the referent group (odds ratio = 1). RESULTS: Of 32,834 subjects, 27,178 had vaginal delivery (operative = 4,908; spontaneous = 22,270) and 5,656 had cesarean delivery. Third- or fourth-degree lacerations occurred in 1,733 (7.8%) women who had spontaneous vaginal delivery compared with 1,098 (22.3%) who had operative vaginal delivery. Overall, 523 women (1.6%) had endometritis. Compared with spontaneous vaginal delivery, primary cesarean delivery with trial of labor conferred a 21.2-fold increased risk of endometritis (95% CI 15.4, 29.1). Even without trial of labor, women after primary cesarean delivery were 10.3 times more likely to develop endometritis (95% CI 5.9, 17.9) than after spontaneous vaginal delivery. The risk of transfusion was highest in women delivered by primary cesarean after labor, 4.2 times higher (95% CI 1.8, 10.1) than spontaneous vaginal delivery. The risk of pneumonia was 9.3 times higher (95% CI 3.4, 25.6) after repeat cesarean delivery with labor. Deep venous thromboses occurred in 15 (0.1%) after spontaneous vaginal delivery, 2 (0.04%) after operative vaginal delivery, and 12 (0.2%) after cesarean delivery. CONCLUSION: Compared with spontaneous vaginal delivery, cesarean delivery is associated with increased risks of endometritis, the need for transfusion, and pneumonia; however, these rates are lower than reported previously. PMID- 15121565 TI - Leukocyte adhesion molecules and reactive oxygen species in preeclampsia. AB - OBJECTIVE: The aim of our study was to compare the expression of leukocyte adhesion molecules, intracellular reactive oxygen species, and vasoactive substances in preeclampsia and matched normotensive pregnancies and to explore differences between pregnancy and the nonpregnant state regarding these parameters. METHODS: Flow cytometry was used to analyze the monocyte and granulocyte expression of adhesion molecules from 20 matched pairs of preeclampsia/normotensive pregnancies and 12 nonpregnant subjects. Basal levels of CD11b, CD11c, CD62L, and CD14 were measured. In addition, expression of human lymphocyte antigen-DR, CD4, CD8, and CD4/CD8 ratio were assessed. Basal reactive oxygen species levels, as well as reactivity upon in vitro stimulation with phorbol 12-myristate 13-acetate, were measured in monocytes and granulocytes with the probes dihydroethidium, dichlorofluorescein-diacetate, and dihydrorhodamine 123. Further, the plasma levels of endothelin-1, the nitric oxide metabolites nitrite/nitrate, and total antioxidant status were analyzed. RESULTS: Monocytes expressed significantly higher levels of CD11b and CD14 in preeclamptic patients compared with normotensive pregnant subjects, whereas CD11c was elevated on both monocytes and granulocytes in pregnancy compared with the nonpregnant state. Both monocytes and granulocytes displayed higher basal, as well as phorbol 12 myristate 13-acetate-stimulated, amounts of reactive oxygen species in the preeclampsia group compared with the normotensive group. We also found the endothelin-1 and antioxidant levels significantly elevated in preeclampsia patients compared with normotensive subjects, whereas no differences were seen between the groups regarding nitrite/nitrate levels. CONCLUSION: These results show that the maternal blood leukocytes are activated in preeclampsia and support the view that oxidative stress is a contributing factor in the pathophysiology of preeclampsia. PMID- 15121566 TI - Rofecoxib versus magnesium sulfate to arrest preterm labor: a randomized trial. AB - OBJECTIVE: To compare oral rofecoxib with intravenous magnesium sulfate as a tocolytic. METHODS: This was a randomized study of patients who were between 22 and 34 weeks of gestation with preterm labor. Patients were randomly assigned to receive either daily oral rofecoxib (50 mg) or intravenous magnesium sulfate for a maximum of 48 hours. Outcome variables included delay of delivery for 48 hours and the incidence of side effects. Data were analyzed by using the Student t test, Mann-Whitney U test, chi(2) test, and repeated-measures analysis of variance. Sample size calculations were based on previous studies of tocolytic efficacy. RESULTS: Two hundred fourteen patients were randomly assigned (105 received rofecoxib and 109 received magnesium sulfate). Delivery was delayed for 48 hours in 95 (90.4%) and 96 (88%) of the patients in the rofecoxib and magnesium sulfate groups, respectively (relative risk 0.97; 95% confidence interval 0.89, 1.06). To show a statistically significant benefit in delay of delivery past 48 hours, a total of 2,686 patients would be required in each group. There was no difference between the groups over the course of the study in cervical dilatation, amniotic fluid index, or cervical length by vaginal ultrasonography. The median hospital days on the original admission were also similar at 2 for both groups (P =.10). There was a higher reported incidence of maternal side effects in the magnesium sulfate group (relative risk 1.81; 95% confidence interval 1.07, 3.06). There was no difference in the incidence of neonatal side effects. CONCLUSION: There was no difference between oral rofecoxib and intravenous magnesium sulfate in arresting preterm labor. PMID- 15121567 TI - Platelet activation, sympathetic tone, and plasma volume in nulligravid women of reproductive age. AB - OBJECTIVE: Preeclampsia is associated with increased platelet activation, increased sympathetic activity, and decreased plasma volume. We sought to estimate the relationship of plasma volume, sympathetic activity, or both to platelet activation in nonpregnant nulligravid women. METHODS: We studied 37 healthy nulligravid subjects during the follicular phase of the menstrual cycle. After intravenous access was obtained, subjects rested in the supine position for 15 minutes. Blood was drawn without venous constriction for measurement of plasma catecholamines (epinephrine and norepinephrine) and complete blood count. Antigenic markers of platelet activation, CD63 and CD61-CD14 (platelet-monocyte aggregates), were measured with flow cytometry. Plasma volume was estimated in the supine position by using Evans blue dye and is expressed in milliliters and corrected for body mass index (BMI). We compared data from the lowest plasma volume/BMI quartile with the 2 middle quartiles combined and with the upper quartile. Data are expressed as mean +/- standard deviation. P <.05 was considered significant. RESULTS: Subjects were aged 26.5 +/- 5.0 years, BMI was 24.0 +/- 3.0 kg/m(2), and plasma volume was 2,685 +/- 429 mL. We identified no significant relationship of platelet concentration to plasma volume/BMI between quartile groups (P =.944). However, there was a significant difference between quartiles for %CD63 expression (P =.013) and for CD61/CD14 expression (P =.018), with the lowest quartile demonstrating elevated platelet activation. CONCLUSIONS: We found evidence that enhanced platelet activation is associated with reduced plasma volume, but not with plasma catecholamine concentrations. There was no association of platelet concentration with reduced plasma volume. We speculate that elements of the clinical syndrome of preeclampsia coexist as a subclinical phenotype before pregnancy. PMID- 15121568 TI - Vasa previa: the impact of prenatal diagnosis on outcomes. AB - OBJECTIVE: To evaluate outcomes and predictors of neonatal survival in pregnancies complicated by vasa previa and to compare outcomes in prenatally diagnosed cases of vasa previa with those not diagnosed prenatally. METHODS: We performed a multicenter study of 155 pregnancies complicated by vasa previa. Cases were obtained from the Vasa Previa Foundation and 6 large hospitals. Comparisons were made between groups based on prenatal diagnosis status and neonatal survival. RESULTS: The overall perinatal mortality was 36% (55 of 155). In 61 cases (39%), vasa previa was diagnosed prenatally; 59 of 61 (97%) infants from these pregnancies survived compared with 41 of 94 (44%) in cases not diagnosed prenatally (P <.001). Median 1- and 5-minute Apgar scores in cases diagnosed prenatally were 8 and 9, respectively, compared with 1 and 4 among survivors in cases not diagnosed prenatally (P <.001). More than half (24 of 41) of surviving neonates born to women without prenatal diagnosis required blood transfusions compared with 2 of 59 diagnosed prenatally (P <.001). Multivariable logistic regression analysis showed that the only significant predictors of neonatal survival were prenatal diagnosis (P <.001) and gestational age at delivery (P =.01). CONCLUSIONS: Good outcomes with vasa previa depend primarily on prenatal diagnosis and cesarean delivery at 35 weeks of gestation or earlier should rupture of membranes, labor, or significant bleeding occur. PMID- 15121569 TI - Paracervical block in incomplete abortion using manual vacuum aspiration: randomized clinical trial. AB - OBJECTIVE: To estimate the effectiveness of paracervical block in controlling pain among women treated with manual vacuum aspiration for an incomplete abortion METHODS: A randomized clinical trial was conducted at Nuestra Senora de Altagracia, a maternal and perinatal referral hospital in the Dominican Republic. The sample size was based on a clinical difference of 1.5 points in the level of pain measured with the visual analog scale using 90% power and a sampling error of 0.04. Women who were at 12 weeks of gestation or less with an incomplete abortion were eligible to participate. They were randomly assigned to receive either the standard treatment of care (manual vacuum aspiration for uterine evacuation with psychological support but no paracervical block) or manual vacuum aspiration treatment with psychological support and paracervical block using 1.0% lidocaine. Patients with active infections, severe illnesses, psychiatric disorders, or allergies to lidocaine were excluded. Intraoperative pain as reported by the women and as documented by an external observer was measured. RESULTS: Although the paracervical block technique used showed a slight reduction in severe pain, there were no clinically or statistically significant differences in intraoperative pain between the 2 groups (relative risk 0.73; 95% confidence interval 0.43, 1.23) with 50% of all patients registering 7 or higher score on a visual analog pain scale of 0-10. However, statistically significant differences were found in each group when comparing the level of preoperative and intraoperative pain described by the patient (P <.001). The manual vacuum aspiration technique and the paracervical block were not accompanied by complications. CONCLUSION: The paracervical block technique used in this study along with psychological support was comparable with pain control using psychological support alone; neither pain management regimen provided sufficient pain control. It is recommended that randomized comparative studies be designed to determine the effectiveness of other paracervical block techniques and the efficacy of the use of analgesics in patients suffering from incomplete abortion treated with manual vacuum aspiration. PMID- 15121570 TI - Comparison of adolescent and young adult self-collected and clinician-collected samples for human papillomavirus. AB - OBJECTIVE: To examine the concordance between self-collected and clinician collected samples for human papillomavirus (HPV) DNA. METHODS: Sexually active adolescent and young adult women aged 14-21 years (N = 101) were enrolled in a prospective cohort study of HPV testing. Participants self-collected vaginal samples for HPV DNA, and clinicians collected cervicovaginal samples for HPV DNA and a cervical cytology specimen. We determined concordance between the results of self- and clinician-collected specimens using a kappa statistic and McNemar's test. RESULTS: Of the 51% of participants who were HPV positive, 53% had 1 type, 25% had 2 types, and 22% had 3 types or more; 25 different HPV types were identified. Self-collected samples detected more participants with HPV than clinician-collected samples (45% versus 42%, P =.65). When results were categorized into presence or absence of high-risk HPV types, agreement between self- and clinician-collected specimens was high (kappa 0.72) and the difference between test results was not significant (McNemar's P =.41). However, when all HPV types detected were considered, agreement was perfect in only 51% of those with 1 or more types of high-risk HPV type. There was no association between agreement and age or HPV type. CONCLUSION: Self testing for HPV DNA may be sufficiently sensitive for the detection of high-risk HPV DNA among adolescent and young adult women in clinical settings. PMID- 15121571 TI - Premenstrual syndrome as a predictor of menopausal symptoms. AB - OBJECTIVE: To estimate whether premenstrual syndrome (PMS) predicts common menopausal symptoms assessed longitudinally for 5 years among women in the transition to menopause. METHODS: Data were obtained from a structured interview questionnaire, daily symptom ratings, and standard measures of depressive symptoms and sleep quality at 7 assessment periods in a population-based cohort of 436 women. Menstrual status was determined by menstrual bleeding dates. Hormones were measured in the early follicular phase, with a maximum of 14 measures per subject. Multivariate logistic regression models for repeated measures were used to estimate the effects of study variables. RESULTS: Premenstrual syndrome significantly decreased with age (P <.001) and with changes in menstrual bleeding status (P =.003). Women with PMS at enrollment were more likely over the 5-year period to report menopausal hot flushes (odds ratio [OR] 2.09; confidence interval [CI] 1.42, 3.08; P <.001); depressed mood (OR 2.34; CI 1.60. 3.43; P <.001); poor sleep (OR 1.72; CI 1.16, 2.53; P =.007), and decreased libido (OR 1.54; CI 1.06, 2.24; P =.024) after adjusting for age, race, diagnosis of major depression, and estradiol. Subjects' fluctuations in estradiol were significantly associated with hot flushes, depressive symptoms, and poor sleep. CONCLUSION: Premenstrual syndrome decreased in the transition to menopause. Women who reported PMS at baseline were at greater risk of menopausal hot flushes, depressed mood, poor sleep, and decreased libido. Further studies of the associations of symptoms and changes in ovarian function are needed to elucidate the underlying symptom physiology and aid in the development of effective treatments for women during the menopausal transition. PMID- 15121572 TI - Professional liability and other career pressures: impact on obstetrician gynecologists' career satisfaction. AB - OBJECTIVE: To investigate the impact of career pressures on career satisfaction and satisfaction with job-specific activities among obstetrician-gynecologists. METHODS: A questionnaire was sent to 1,500 member-Fellows of The American College of Obstetricians and Gynecologists in June 2001. The analyses were designed to examine the relationship between career pressures in 3 domains on clinicians' professional satisfaction. RESULTS: Overall career satisfaction and satisfaction with job-specific activities were both inversely related to the perceived impact of career pressures. The major impact reported was that liability insurance costs would shorten the duration of the members' careers. Managed care had less impact than liability, with moderate concern surrounding the limitation of diagnostic and treatment options. Obstetrician-gynecologists were less satisfied with their careers and job-specific activities if they believed the cost or time of obtaining continuing medical education requirements to be a burden. CONCLUSION: Career pressures produced by liability insurance costs have more negative impact on clinicians' satisfaction with their professional lives and job-specific activities than managed care and requirements for continuing medical education. PMID- 15121573 TI - Suture closure of subcutaneous fat and wound disruption after cesarean delivery: a meta-analysis. AB - OBJECTIVE: To define the role of suture closure of the subcutaneous dead space in preventing wound complications after cesarean delivery. DATA SOURCES: We searched MEDLINE, the Cochrane Database of Systematic Reviews, and the bibliographies of major texts and review articles. METHODS OF STUDY SELECTION: Only studies in which patients undergoing cesarean delivery were randomly assigned to closure of the subcutaneous space or to no closure were included. Each study was required to report on at least 1 of the following outcomes: wound infection, hematoma, seroma, or separation. The studies also reported "wound disruption," a combination of these outcomes which either explicitly stated or strongly implied the need for further wound care. Six studies meeting criteria were identified. TABULATION, INTEGRATION, AND RESULTS: Three studies included 875 patients with any subcutaneous thickness and noted a decrease in wound disruption with closure (relative risk [RR] 0.56; 95% confidence interval [CI] 0.36, 0.86). Two studies reported results from 181 patients with incision depth of 2 cm or less and noted no difference (RR 1.01; 95% CI 0.46, 2.20). Five studies reported results on 887 patients with wound thickness greater than 2 cm. Although only 1 study had a significant effect by itself, when results were combined, there was a significant decrease in wound disruption (RR 0.66; 95% CI 0.48, 0.91). This reduction seems to be largely a result of decreased wound seromas (4 studies, 852 patients, RR 0.42; 95% CI 0.24, 0.75). In women with wound thickness greater than 2 cm, subcutaneous closure resulted in a risk reduction of 6.2%, and 16.2 women would need subcutaneous closure to prevent 1 wound disruption (number needed to treat). CONCLUSION: Suture closure of subcutaneous fat during cesarean delivery results in a 34% decrease in risk of wound disruption in women with fat thickness greater than 2 cm. PMID- 15121574 TI - Diagnosis, controversies, and management of the syndrome of hemolysis, elevated liver enzymes, and low platelet count. AB - Hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome has been recognized as a complication of preeclampsia-eclampsia for decades. Recognition of this syndrome in women with preeclampsia is increasing because of the frequency of blood test results that reveal unexpected thrombocytopenia or elevated liver enzymes. The diagnosis of HELLP syndrome requires the presence of hemolysis based on examination of the peripheral smear, elevated indirect bilirubin levels, or low serum haptoglobin levels in association with significant elevation in liver enzymes and a platelet count below 100,000/mm(3) after ruling out other causes of hemolysis and thrombocytopenia. The presence of this syndrome is associated with increased risk of adverse outcome for both mother and fetus. During the past 15 years, several retrospective and observational studies and a few randomized trials have been published in an attempt to refine the diagnostic criteria, to identify risk factors for adverse pregnancy outcome, and to treat women with this syndrome. Despite the voluminous literature, the diagnosis and management of this syndrome remain controversial. Recent studies suggest that some women with partial HELLP syndrome may be treated with expectant management or corticosteroid therapy. This review will emphasize the controversies surrounding the diagnosis and management of this syndrome. Recommendation for diagnosis, management, and counseling of these women is also provided based on results of recent studies and my own clinical experience. PMID- 15121575 TI - Complications associated with global endometrial ablation: the utility of the MAUDE database. PMID- 15121577 TI - Complications associated with global endometrial ablation: the utility of the MAUDE database. PMID- 15121578 TI - Foley balloon to tamponade bleeding in the retropubic space. PMID- 15121580 TI - Outcome of fertility-sparing treatment with progestins in young patients with endometrial cancer. PMID- 15121581 TI - Package insert for rhesus immune globulin. PMID- 15121583 TI - First-trimester sonographic screening for Down syndrome. PMID- 15121584 TI - First-trimester sonographic screening for Down syndrome. PMID- 15121586 TI - Asthma during pregnancy. PMID- 15121588 TI - Intrapartum computerized fetal heart rate parameters and metabolic acidosis at birth. PMID- 15121590 TI - Vaginal versus cesarean delivery for breech presentation in California: a population-based study. PMID- 15121591 TI - Acyclovir prophylaxis to prevent herpes simplex virus recurrence at delivery: a systematic review. PMID- 15121594 TI - Helicobacter pylori infection and dyspepsia in pregnancy. PMID- 15121596 TI - ACOG Committee Opinion. Number 294, May 2004. At-risk drinking and illicit drug use: ethical issues in obstetric and gynecologic practice. AB - Abuse of alcohol and drugs is a major health problem for American women across differences in socioeconomic status, race, ethnicity, and age, and it is costly to individuals and to society. Obstetrician-gynecologists have an ethical obligation to learn and use a protocol for universal screening questions, brief intervention, and referral to treatment in order to provide patients and their families with medical care that is state-of-the-art, comprehensive, and effective. In this Committee Opinion, the American College of Obstetricians and Gynecologists' Committee on Ethics proposes an ethical rationale for this protocol in both obstetric and gynecologic practice and provides guidelines for resolving common ethical dilemmas related to drug and alcohol use that arise in the clinical setting. PMID- 15121597 TI - Abdominal sacral colpopexies complicated by vaginal graft extrusion. PMID- 15121598 TI - Entero mesh vaginal fistula secondary to abdominal sacral colpopexy. AB - BACKGROUND: Abdominal sacral colpopexy is a popular method for resupporting the vaginal apex. Bleeding and infection are the most common complications. We report a complication resulting in a small bowel fistula. CASE: A 48-year-old woman developed a chronic vaginal discharge 4-6 months after routine abdominal sacral colpopexy in which a velour mesh remained exposed in the pelvis. Conservative measures failed to control the intermittent copious discharge from the upper vaginal vault where the mesh was visualized. At laparotomy, an entero mesh vaginal fistula was discovered. Excellent long-term results were obtained by removal of the mesh along with resection of the involved small intestine. CONCLUSION: At the time of abdominal sacral colpopexy, we recommend that mesh not remain exposed in the pelvis. PMID- 15121599 TI - Abdominal sacral colpopexy mesh erosion resulting in a sinus tract formation and sacral abscess. AB - BACKGROUND: Complications associated with the use of synthetic mesh during an abdominal sacral colpopexy procedure include mesh infection and erosion into the vaginal vault and sacral osteomyelitis. CASE: This case report describes the management of an abdominal sacral colpopexy procedure that was complicated by postoperative vaginal mesh erosion, formation of a fistulous tract from the vaginal apex to the sacrum, and development of diskitis, osteomyelitis, and a sacral abscess. CONCLUSION: Treatment of a vaginal mesh erosion complicated by the formation of a sinus tract after abdominal sacral colpopexy should include extensive sinus tract resection in addition to complete mesh removal. PMID- 15121600 TI - Transvaginal endoscopic removal of eroded mesh after abdominal sacral colpopexy. AB - BACKGROUND: Abdominal sacral colpopexy with permanent mesh has become the preferred abdominal technique for correcting posthysterectomy vaginal vault prolapse. Although rare, mesh erosion after sacral colpopexy is often challenging to manage. We report on 3 cases of mesh erosion after abdominal sacral colpopexy managed by transvaginal endoscopic removal of the mesh. CASES: The cases involve patients who underwent an abdominal sacral colpopexy and had vaginal mesh erosions within 3 years of their surgeries. Conservative and traditional transvaginal techniques failed, and the patients continued to complain of vaginal discharge. All patients underwent transvaginal endoscopic removal of the mesh and are cured of their chronic discharges. CONCLUSION: Transvaginal endoscopic removal is an effective, minimally invasive option for removal of eroded mesh after abdominal sacral colpopexy. PMID- 15121601 TI - Brace for the case: be prepared for anaphylaxis. PMID- 15121602 TI - Anaphylaxis from medroxyprogesterone acetate. AB - BACKGROUND: More than 30 million women in more than 90 countries use depot medroxyprogesterone acetate as an injectable contraceptive agent, and it is thought to be very safe. However, it should be recognized that serious and potentially life-threatening adverse effects can occur. CASE: A 40-year-old para 2 went into anaphylactic shock after receiving 150 mg of depot medroxyprogesterone acetate intramuscularly. She was not taking any other medication, and there was no history of allergy to food or cosmetics. She responded fully to immediate resuscitation. A repeat episode occurred when she received another dose 12 weeks later. CONCLUSION: Although thought to be very safe, life-threatening adverse effects can occur with depot medroxyprogesterone acetate, and practitioners should be prepared for such eventuality. PMID- 15121603 TI - A rare drug reaction to methotrexate after treatment for ectopic pregnancy. AB - BACKGROUND: Ectopic pregnancies are commonly diagnosed and treated in physicians' offices. In the hemodynamically stable patient, therapy often includes treatment with methotrexate. Well-known adverse effects of this drug include mucositis, abdominal cramping, and malaise. We report a case of a rare drug reaction after treatment with methotrexate. CASE: A 34-year-old, gravida 2, para 0, at 7 weeks of gestation by last menstrual period was diagnosed with an ectopic pregnancy and treated with methotrexate. The patient had an anaphylactoid reaction shortly after administration of methotrexate. CONCLUSION: Methotrexate is a commonly used therapy for ectopic pregnancies in the outpatient setting. Practitioners should be aware of the potential adverse reactions to methotrexate. PMID- 15121604 TI - Desquamating vaginal mucosa from chlorhexidine gluconate. AB - BACKGROUND: Chlorhexidine gluconate is a commonly used and effective antiseptic agent for preparing patients for surgery. CASE: A healthy premenopausal woman was taken to the operating room for a planned laparoscopically assisted vaginal hysterectomy. After vaginal, vulvar, perineal, and abdominal cleansing with chlorhexidine gluconate, the patient developed a desquamating vaginal reaction that was treated with intravenous corticosteroids, antihistamine, topical conjugated estrogen, and hydrocortisone cream. The planned surgery was aborted, and the patient recovered uneventfully overnight in the postanesthesia care unit. With continued application of conjugated estrogen cream, the patient's vaginal mucosa was well healed within 2 weeks. CONCLUSION: Although chlorhexidine gluconate has been used effectively to minimize surgical site infection in vaginal surgery, the possibility for adverse reaction should be considered. PMID- 15121605 TI - Surgical cytoreduction and hormone therapy of an advanced endometrial stromal sarcoma of the ovary. AB - BACKGROUND: Primary endometrial stromal sarcomas of the ovary are rare gynecologic malignancies. We report a disseminated case of this tumor arising from ovarian endometriosis. CASE: A 45-year-old woman presented with an abdominal pelvic mass and an elevated CA 125. Exploration showed extensive tumor spread from the ovaries to the upper abdomen. Surgery included a total hysterectomy, bilateral salpingo-oophorectomy, splenectomy, partial gastrectomy, partial pancreatectomy, transverse colectomy, appendectomy, and omentectomy. Final pathology showed a low-grade endometrial stromal sarcoma of the ovary arising from foci of endometriosis. Megestrol acetate was initiated, and she is currently without evidence of disease. CONCLUSION: This is an advanced case of a primary low-grade endometrial stromal sarcoma of the ovary arising from endometriosis managed by total resection and progestational therapy. PMID- 15121606 TI - Recombinant factor VIIa in management of spontaneous subcapsular liver hematoma associated with pregnancy. AB - BACKGROUND: Spontaneous subcapsular liver hemorrhage is a rare but life threatening complication of pregnancy. Optimal management of an expanding hematoma or ruptured capsule has not been established. CASES: We report 3 patients with preeclampsia and hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome with spontaneous subcapsular liver hematomas. The first 2 patients with ruptured liver hematomas experienced life-threatening hemorrhage. The third patient experienced uncontrollable vaginal bleeding, liver hemorrhage, and was in imminent danger of capsule rupture. Despite aggressive surgical intervention and traditional blood component therapy, adequate hemostasis could not be achieved in any of these patients. Recombinant factor VIIa was used to achieve hemostasis in all three patients. CONCLUSION: Recombinant factor VIIa is an effective adjunct in the treatment of preeclamptic patients with expanding or ruptured subcapsular liver hematoma. PMID- 15121607 TI - An accessory uterine cavity as a cause of pelvic pain. AB - BACKGROUND: Pelvic pain is a common gynecologic problem. We report a case of a noncommunicating accessory uterine cavity as a cause of pelvic pain in a young adolescent. CASE: A 17-year-old nulligravida presented with worsening pelvic pain. Transvaginal ultrasound revealed a 3-cm cavity within the myometrium. Hysteroscopy revealed a normal uterine cavity. Surgical excision of the accessory cavity just lateral to the normal uterine cavity was achieved through laparotomy. Pathology confirmed endometrial tissue in the accessory cavity. Pelvic pain resolved after surgery. CONCLUSION: A noncommunicating uterine cavity should be considered in the differential diagnosis of pelvic pain. PMID- 15121609 TI - Minimally invasive management of an advanced abdominal pregnancy. AB - BACKGROUND: Advanced abdominal pregnancy is a rare, life-threatening condition that presents a number of challenges. CASE: A 29-year-old primigravida with 10 years of secondary infertility and a previous tuboplasty had a 21-week abdominal pregnancy treated with preoperative arterial embolization before laparoscopically assisted fetal delivery. Postoperatively, 4 cycles of methotrexate were administered at 50 mg/m2 intramuscularly every 3 weeks for the retained abdominal placenta. Subsequent spontaneous conception occurred, and a live, full-term infant was delivered by cesarean delivery 17 months later. No adverse sequelae were found during long-term follow-up. CONCLUSION: This report demonstrates successful minimally invasive management of an advanced abdominal pregnancy with a multimodal approach that included preoperative arterial embolization, laparoscopically assisted delivery, and judicious use of postoperative methotrexate. PMID- 15121608 TI - Stage IV large B cell lymphoma presenting as gigantomastia and pulmonary hypertension. AB - BACKGROUND: Gigantomastia is a rare complication of pregnancy usually associated with benign conditions, including end-organ hypersensitivity to normal hormone levels, penicillamine therapy, mirror syndrome, and benign or glandular fibroadenomas. CASE: A young woman presented at 30 weeks of gestation with chest pain and gigantomastia. Echocardiography performed because of the patient's tachypnea and tachycardia disclosed pulmonary hypertension. After spontaneous delivery, core needle breast and axillary lymph node biopsies and computerized tomography imaging were performed, and stage IV diffuse large B cell lymphoma with infiltration of the breasts was confirmed. The patient responded to systemic chemotherapy with resolution of the gigantomastia and pulmonary artery hypertension. CONCLUSION: This case demonstrates that systemic malignancies such as diffuse large B cell lymphoma should be considered in the differential diagnosis of gigantomastia during pregnancy. In addition, malignancy-related pulmonary hypertension during pregnancy may be reversible after chemotherapy, as reported in nonpregnant patients. PMID- 15121610 TI - Successful one-stage radical removal of intravenous leiomyomatosis extending to the right ventricle. AB - BACKGROUND: Intravenous leiomyomatosis is a rare tumor originating from the uterus that spreads through vessels. This tumor, while histologically benign, can cause fatal cardiovascular symptoms resulting from growth within the heart cavity. CASE: A 51-year-old woman with chronic hypertension was found to have a murmur. Echocardiogram, magnetic resonance imaging, and clinical evaluation revealed an intravenous leiomyomatosis extending to the right ventricle. The appropriate therapy for intravenous leiomyomatosis is complete resection. In this patient, one-stage cardiotomy with laparotomy resulted in successful radical removal of the tumor. CONCLUSION: Our patient underwent successful radical therapy for an intravenous leiomyomatosis extending to the heart, which involved complete resection of the tumor by one-stage cardiotomy with laparotomy. PMID- 15121611 TI - Successful pregnancy after thermal balloon endometrial ablation. AB - BACKGROUND: Thermal balloon ablation is as effective as other electrosurgical modalities used for endometrial ablation. Pregnancy rate after endometrial ablation is 0.24-0.68%. We report a near-term viable pregnancy after thermal balloon uterine ablation. CASE: Thermal balloon uterine endometrial ablation was performed on a 38-year-old woman with menorrhagia. She conceived 11 months after the operation and decided to continue the pregnancy. After 35 weeks of uneventful gestation, she spontaneously delivered a liveborn infant. CONCLUSION: Although rare, pregnancy after endometrial ablation is possible. Obstetric complications, such as pathologic placental adherence and fetal demise due to a small, scarred uterine cavity, have been reported. This pregnancy went to 35 weeks without complication despite a three-chambered appearance of the uterus. PMID- 15121612 TI - A life-threatening event: uterine cervical arteriovenous malformation. AB - BACKGROUND: Uterine cervical arteriovenous malformation is a rare cause of vaginal bleeding. CASE: A 32-year-old multigravida presented with severe vaginal bleeding originating in the cervix, which resulted in a hypovolemic shock. Attempts to control the bleeding included hysterectomy, pelvic arterial embolization, and upper vaginectomy. Each proved unsuccessful. Histopathologic examination revealed an arteriovenous malformation. Despite local packing, suturing of the vault area, and brachytherapy to the vaginal vault, bleeding persisted. Treatment with GnRH agonist and tranexamic acid stopped the bleeding. CONCLUSION: Severe vaginal bleeding can be the result of cervical arteriovenous malformation, and GnRH agonist may be used for treatment. PMID- 15121613 TI - Postpartum uterine arteriovenous fistula. AB - BACKGROUND: Uterine arteriovenous communications are uncommon lesions that may be associated with life-threatening postpartum and postinstrumentation hemorrhage. CASE: A primigravida presented with infected retained products of conception. Excessive hemorrhage of unclear etiology occurred at dilation and curettage. After a second episode of bleeding, the patient received a diagnosis of uterine arteriovenous fistula. CONCLUSION: Uterine arteriovenous communications should be included in the differential diagnosis in patients with excessive postpartum or postinstrumentation bleeding. Color and spectral flow Doppler can aid diagnosis and clinical management. PMID- 15121614 TI - Massive subchorionic hematomas following thrombolytic therapy in pregnancy. AB - BACKGROUND: Medical therapy for thrombosed valve in pregnancy has become an acceptable alternative to surgery, especially in hemodynamically compromised patients. Placental changes after thrombolytic therapy have rarely been reported. CASES: Sonograms were done within 24 hours after administration of thrombolytic agents at 15 and 26 weeks of gestation, respectively, in 2 women whose pregnancies were complicated with thrombosis of prosthetic mitral valves. Both patients developed massive subchorionic hematomas, which persisted in 1 patient who underwent cesarean delivery at 34 weeks of gestation for cardiac indications (Apgar scores 9 and 10 at 1 minute and 5 minutes, respectively). The hematomas resolved in the other patient, who delivered at term. CONCLUSION: Massive subchorionic hematomas may be observed in patients after thrombolytic therapy. Other reports are needed to establish whether such placental findings are common lesions after such therapy and to determine their impact on pregnancy outcome. PMID- 15121615 TI - Relapse after more than 20 years of follow-up for epithelial ovarian carcinoma. AB - BACKGROUND: Very late relapse of ovarian cancer is unusual and may present with atypical symptoms. CASES: We diagnosed 3 cases of relapse occurring after more than 20 years of follow-up. In the first case, the first recurrence was diagnosed by an appendicitis syndrome. With the second recurrence, small pelvic nodules were detected by fluorodeoxyglucose scintigraphy, whereas other imaging method results were negative. In the second case, a nodule in the axilla revealed the recurrence, and imaging methods confirmed multiple metastasis. In the third case, the patient presented with paroxysmal abdominal pain, and fluorodeoxyglucose scintigraphy showed a tiny lesion. CONCLUSION: Late relapses of ovarian cancer raise the issue of regrowth of dormant cells or the development of a new primary cancer. The absence of family history and BRCA gene mutations in these 3 patients favor late recurrence. Fluorodeoxyglucose scintigraphy was useful for diagnosis, particularly of small lesions not visible by classic imaging methods. PMID- 15121616 TI - Ketanserin in women with chronic hypertension and underlying thrombophilia. AB - BACKGROUND: Pregnant women with chronic hypertension and thrombophilia have an increased risk for preeclampsia and/or intrauterine growth restriction (IUGR). Ketanserin lowers blood pressure and inhibits serotonin-induced platelet aggregation. CASES: A 38-year-old woman with chronic hypertension had a first pregnancy with severe hemolysis, elevated liver enzymes, and low platelets (HELLP) syndrome. A second pregnancy with prophylactic oral ketanserin and low dose aspirin was uneventful. The third pregnancy without oral ketanserin was again complicated by severe superimposed preeclampsia and IUGR. During this pregnancy a prothrombin mutation was found. A 37-year-old woman with chronic hypertension developed severe early-onset superimposed preeclampsia in her first pregnancy. In the 3 consecutive pregnancies, she was treated with low-molecular weight heparin because of a factor XII deficiency. Only the pregnancy with the use of prophylactic oral ketanserin was uncomplicated. CONCLUSION: Clinical trials are warranted to analyze the role of oral ketanserin in preventing preeclampsia and IUGR. PMID- 15121617 TI - Laparoscopic removal of a large adnexal mass. AB - BACKGROUND: Laparoscopic removal of adnexal masses has been facilitated by the use of commercially available retrieval bags. Masses up to 18 cm can be removed using zipper storage bags. Masses with greater diameters have required laparotomy or techniques that do not isolate the mass from the peritoneal cavity. By using larger bags available in the operative suite, even the very large adnexal mass can be isolated from the peritoneal cavity and extracted via laparoscopy. CASE: A 46-year-old nulligravida presented with a 20.9-cm adnexal mass. She underwent laparoscopy and extraction of a dermoid with an improvised retrieval bag without intraperitoneal spillage. CONCLUSION: Very large adnexal masses can be removed laparoscopically without intraperitoneal spillage despite the lack of commercially available bags for this purpose. PMID- 15121618 TI - Spontaneous rupture of the uterine vessels in pregnancy. AB - BACKGROUND: Hemoperitoneum resulting from spontaneous rupture of the uterine vessels in pregnancy is rare and associated with high maternal and fetal mortality. CASE: A woman presented with acute abdominal pain and hypovolemic shock at 20 weeks of gestation. Immediate laparotomy revealed massive hemoperitoneum resulting from spontaneous rupture of the left uterine vessels associated with a left adnexal mass consisting of decidualized endometriosis. The fetus was delivered by hysterotomy, hemostasis was achieved, and the woman made a good recovery. CONCLUSION: We report a case of hemoperitoneum in pregnancy that resulted from spontaneous rupture of the uterine vessels associated with decidualized endometriosis. PMID- 15121619 TI - Mycophenolate mofetil in pregnancy after renal transplantation: a case of major fetal malformations. AB - BACKGROUND: Mycophenolate mofetil has teratogenic properties in rats and rabbits. Previous human studies have reported an increased rate of fetal losses with its use. We report a case of major fetal malformations due to mycophenolate mofetil. CASE: The patient was treated with mycophenolate mofetil before conception and during the first trimester of pregnancy. The fetus had multiple malformations, specifically, facial dysmorphology and midline anomalies, including agenesis of the corpus callosum. CONCLUSION: This case of fetal malformation attributable to mycophenolate mofetil must be taken into consideration when considering pregnancy in an organ-transplant recipient. PMID- 15121620 TI - Tumor-level serum testosterone associated with human immunodeficiency virus lipodystrophy syndrome. AB - BACKGROUND: Human immunodeficiency virus (HIV) lipodystrophy syndrome consists of insulin resistance, hyperlipidemia, increased waist-to-hip ratios, and relative hyperandrogenemia, which resembles polycystic ovary syndrome. CASE: A 30-year-old HIV-positive multipara with lipodystrophy syndrome had an elevated level of serum total testosterone (244 ng/dL) in the tumor range. CONCLUSION: In HIV lipodystrophy syndrome, marked elevations of total serum testosterone in the range reserved for androgen-producing tumors may occur. The elevated testosterone levels are derived from the ovary and are responsive to gonadotropin suppression. PMID- 15121621 TI - Use of a hemoglobin-based oxygen carrier in the treatment of severe anemia. AB - BACKGROUND: Hemoglobin-based oxygen carriers hold promise for the treatment of acute anemia. CASE: We report a patient with severe dysfunctional uterine bleeding. During her hospitalization, her lowest hemoglobin level was 3.1 g/dL, with a hematocrit of 9.3%. An investigational product, o-raffinose cross-linked human hemoglobin solution (hemoglobin raffimer), was infused along with ongoing high-dose recombinant human erythropoietin and estrogen. The time until the patient's own hematopoiesis provided sufficient red blood cell mass was successfully managed by reducing oxygen demand and providing multiple hemoglobin based oxygen carrier infusions. After hemoglobin-based oxygen carrier administration, transient pulmonary hypertension and fever were noted. She was discharged after corrective surgery 7 days after hemoglobin-based oxygen carrier administration with a hemoglobin level of 7.8 g/dL. CONCLUSION: The hemoglobin level-based oxygen carrier improved oxygen delivery and permitted uterine corrective surgery. PMID- 15121622 TI - Multiple vaginal wall cysts: diagnosis and surgical management. AB - BACKGROUND: Simple cysts of the female genital tract may become symptomatic and require surgical removal. CASE: A 20-year-old woman had complaints of a vaginal bulge, pelvic pressure, dyspareunia, and stress urinary incontinence. Two cystic masses were seen in the vagina. A magnetic resonance imaging scan revealed 2 additional larger cysts. Urodynamic evaluation was significant for stress incontinence at 200 mL and a Valsalva leak-point pressure of 51 cm H(2)0. The 2 smaller cysts were removed intact through a superficial incision. The larger cysts were intentionally ruptured and dissected out. Histologic examination revealed a low cuboidal or a ciliated columnar epithelium that stained positive with mucicarmine, consistent with mullerian origin. One year after surgical excision, the patient remained symptom free with no urinary incontinence and no recurrence of the cysts. CONCLUSION: Magnetic resonance imaging was useful in delineating the course and anatomic arrangement of the vaginal cysts, but more importantly, identified another cyst that was not readily apparent at physical examination. PMID- 15121623 TI - Intravenous and inhaled epoprostenol for primary pulmonary hypertension during pregnancy and delivery. AB - BACKGROUND: Primary pulmonary hypertension carries a significant mortality risk during pregnancy and delivery. CASE: A 36-year-old pregnant woman with primary pulmonary hypertension was transferred to us with severe dyspnea. Intravenous epoprostenol was started, titrated, and maintained until labor augmentation. Because systemic epoprostenol treatment can interfere with platelet aggregation, we switched to inhaled epoprostenol, administered under a U.S. Food and Drug Administration-approved investigational new drug license, before epidural catheter placement. The inhaled drug was continued because it achieved better control of pulmonary hypertension. An uneventful forceps-assisted vaginal delivery was performed, and intravenous epoprostenol was restarted after the delivery. Mother and baby were well 6 months postpartum. CONCLUSION: Intravenous epoprostenol treatment is effective in management of pregnant and postpartum women with primary pulmonary hypertension. Inhaled epoprostenol was effective during the intrapartum and immediate postpartum period. PMID- 15121624 TI - Conservative surgery for uterine incisional necrosis complicating cesarean delivery. AB - BACKGROUND: Hysterectomy is the usual management of uterine incisional necrosis with dehiscence as a complication of cesarean delivery. Debridement with resuture of the uterine wound as an alternative therapy would conserve fertility at the risk of continued necrosis and infection. Unfortunately the literature provides few guidelines in deciding when to select conservative surgery and whether it is a safe option. CASES: Three cases are presented with uterine incisional necrosis complicating cesarean delivery. In 2 instances, uterine conservation was considered an acceptable option and was successfully completed. The third patient underwent hysterectomy because of clinical features deemed to be contraindications to conservative surgery. CONCLUSION: In well-selected patients, debridement and resuture of the uterine incision is an acceptable alternative to hysterectomy for uterine incisional necrosis. PMID- 15121625 TI - Immature ovarian teratoma with hyponatremia and low antidiuretic hormone level. AB - BACKGROUND: Inappropriate antidiuretic hormone secretion syndrome is rare in patients with gynecologic tumors. CASE: A 22-year-old woman presented with inappropriate antidiuretic hormone secretion symptoms during the 2 months preceding the diagnosis of an immature ovarian teratoma. Vasopressin levels in serum and in the urine were very low. Restriction of water intake and surgical removal of the teratoma resulted in the definitive correction of the hyponatremia. This observation suggests that immature teratoma cells can produce a vasopressin-like factor, and the syndrome may be a sign of an ovarian malignancy. CONCLUSION: Pelvic organs should be examined when the more common causes of inappropriate antidiuretic hormone secretion syndrome have been ruled out. PMID- 15121626 TI - Aortic dissection in pregnancy. AB - BACKGROUND: Aortic dissection is a rare but life-threatening disease. The most common predisposing cause is chronic hypertension. CASE: A 38-year-old multipara with chronic hypertension was admitted at 38 weeks of gestation reporting new onset severe chest pain. Cardiac enzymes, blood gases, electrocardiogram, and chest radiograph were normal. The chest pain and the hypertension worsened despite treatment with narcotics. Bedside transesophageal echocardiography revealed aortic dissection. Uneventful cesarean delivery was followed by surgical repair of the aorta. CONCLUSION: Aortic dissection should be considered when a pregnant woman presents with preeclampsia superimposed on chronic hypertension and intractable chest pain. PMID- 15121627 TI - Nitroglycerin for relaxation to establish a fetal airway (EXIT procedure). AB - BACKGROUND: The ex utero intrapartum treatment (EXIT) procedure is a technique designed to establish an airway at the time of delivery in fetuses at risk of airway obstruction and requires maintenance of uterine relaxation to continue placental perfusion and prevent placental separation. We describe the use of intravenous nitroglycerin to maintain uterine relaxation during the EXIT procedure. CASE: A 17-year-old primigravida with a fetus known to have an anterior neck mass was admitted for a scheduled operative delivery at 38 weeks of gestation using a modified EXIT procedure. Anesthesia was administered with a combined spinal-epidural technique. Intravenous nitroglycerin was administered as a bolus and then as a continuous infusion to maintain uterine relaxation until evaluation of the neonatal airway was completed. CONCLUSION: Intravenous nitroglycerin is an effective agent for maintenance of uterine relaxation and placental perfusion during the EXIT procedure. PMID- 15121628 TI - Very-early-onset discordant growth in monochorionic twin pregnancy. AB - BACKGROUND: The significance of growth restriction in the first trimester in karyotypically normal fetuses is uncertain. CASE: We report a case of diamniotic monochorionic twin pregnancy with marked growth discordance noted in the first trimester. No major congenital or karyotype abnormalities or ongoing evidence of twin-twin transfusion syndrome were found during gestation. Birth weight discordance in the twins was 49.5%. Histologic examination revealed immature villi, with a shortage of terminal villi and an abundance of intermediate mature villi, as well as hypoxic areas with altered villi in the part of the placenta supplying the smaller twin. CONCLUSION: The causes underlying discordant growth in karyotypically normal twin pregnancies without evidence of twin-twin transfusion syndrome are not entirely clear. There may be other conditions responsible for discordant growth that occur in the first or early second trimester of pregnancy. PMID- 15121629 TI - Management of Eisenmenger syndrome in pregnancy with sildenafil and L-arginine. AB - BACKGROUND: Eisenmenger syndrome in pregnancy may be a life-threatening disease despite recent additions to the treatment options. CASE: We present a woman with severe pulmonary hypertension due to Eisenmenger syndrome treated during pregnancy and delivery and postpartum with L-arginine and sildenafil to enhance the nitric oxide pathway. This combination was associated with significant improvement in the mother's clinical and hemodynamic condition and fetal well being. CONCLUSION: The concomitant use of sildenafil and L-arginine for the management of pulmonary hypertension in pregnancy, combined with multidisciplinary care, permitted a good outcome for the mother and her infant. PMID- 15121631 TI - A menopausal woman with mullerian agenesis, a leiomyoma, an inguinal hernia, and cystadenofibromas. AB - BACKGROUND: Mullerian agenesis, the second most common cause of primary amenorrhea, affects 1 in 4,000 to 10,000 women. Although the majority of these women have complete absence of the uterus, a small percentage can have a rudimentary uterus. CASE: A menopausal nulligravida with known mullerian agenesis developed a large pelvic mass and an inguinal hernia. The patient underwent an exploratory laparotomy, which revealed a large leiomyoma growing from a small rudimentary uterus, an indirect inguinal hernia, and bilateral benign serous cystadenofibroma. CONCLUSION: Women with mullerian agenesis and rudimentary uteri can develop gynecologic pathology similar to women with normal genital tracts. PMID- 15121630 TI - Stage IIIC small cell carcinoma of the ovary: survival with conservative surgery and chemotherapy. AB - BACKGROUND: Small cell carcinoma of the ovary is an aggressive tumor primarily affecting young women. Despite adjuvant therapy, the majority of patients described in the literature have fared poorly, even when the disease is diagnosed at an early stage. CASE: A 19-year-old nulligravida with small cell carcinoma of the left ovary underwent conservative surgery with staging and was found to have stage IIIC disease. She received multiagent chemotherapy with vinblastine, cisplatin, cyclophosphamide, bleomycin, doxorubicin, and etoposide and is alive and doing well more than 2 years after completion of her therapy, with no evidence of disease. CONCLUSION: In young patients who desire future fertility, conservative surgery followed by aggressive multiagent chemotherapy may be an effective treatment regimen and warrants further consideration. PMID- 15121632 TI - Anatomical reconstruction after vulvectomy. AB - BACKGROUND: Vulvectomy is a disfiguring operation detrimental to self-esteem and female identity, but surgical techniques for anatomical reconstruction are not routinely performed. CASES: Anatomical reconstruction of the vulva is illustrated in 1) a patient with superficial vulvectomy, 2) a patient with deep anterior vulvectomy, and 3) a patient with extended radical vulvectomy. Random skin flaps and axial-pattern skin flaps based on the deep external and the internal pudendal arteries were used. All patients had good postoperative results. CONCLUSION: Established reconstructive procedures can be used to restore the labial folds, vestibulum, posterior commissure, and perineum. We advocate reconstruction of the anterior commissure with clitoral protrusion after superficial and deep vulvectomy. PMID- 15121633 TI - Prolonged retention of laminaria fragments: a rare complication of induced abortion. AB - BACKGROUND: Laminaria are used worldwide for the preparation of the cervix for termination of pregnancy. CASE: The patient presented with recurrent episodes of pelvic inflammatory disease. Her history was significant for treatment with laminaria 4 years before presentation. After 6 months of evaluation, a diagnostic dilation and curettage was performed with removal of laminaria fragments. CONCLUSION: Evaluation for complete removal of laminaria is critical. Retention of laminaria should be considered in the workup of patients who have pelvic inflammatory disease or genital infections and have received laminaria in the past. PMID- 15121634 TI - Uterine cervical diverticulum resembling a degenerated leiomyoma. AB - BACKGROUND: Uterine diverticulum is a very rare anomaly. Most previously reported cases were called "pregnancy-associated sacculations," and only 2 cases were considered true uterine diverticula. CASE: A 41-year-old woman presented with fever and lower abdominal pain. An extrauterine mass was detected, and a hysterectomy was performed. Pathological examination revealed a cystic uterine diverticulum lined with cervical glands and myometrium arising from the cervix. The final diagnosis was an infected uterine cervical diverticulum. CONCLUSION: Cervical diverticulum is a rare lesion that should be added to the differential diagnosis of a woman presenting with a pelvic mass. PMID- 15121635 TI - Influenza virus inhibits ENaC and lung fluid clearance. AB - Fluid-free alveolar space is critical for normal gas exchange. Influenza virus alters fluid transport across respiratory epithelia producing rhinorrhea, middle ear effusions, and alveolar flooding. However, the mechanism of fluid retention remains unclear. We investigated influenza virus strain A/PR/8/34, which can attach and enter mammalian cells but is incapable of viral replication and productive infection in mammalian epithelia, on epithelial sodium channels (ENaC) in rat alveolar type II (ATII) cells. In parallel, we determined the effects of virus on amiloride-sensitive (i.e., ENaC-mediated) fluid clearance in rat lungs in vivo. Although influenza virus did not change the inulin permeability of ATII monolayers, it rapidly reduced the net volume transport across monolayers. Virus reduced the open probability of single ENaC channels in apical cell-attached patches. U-73122, a phospholipase C (PLC) inhibitor, and PP2, a Src inhibitor, blocked the effect of virus on ENaC. GF-109203X, a protein kinase C (PKC) inhibitor, also blocked the effect, suggesting a PKC-mediated mechanism. In parallel, intratracheal administration of influenza virus produced a rapid inhibition of amiloride-sensitive (i.e., ENaC-dependent) lung fluid transport. Together, these results show that influenza virus rapidly inhibits ENaC in ATII cells via a PLC- and Src-mediated activation of PKC but does not increase epithelial permeability in this same rapid time course. We speculate that this rapid inhibition of ENaC and formation of edema when the virus first attaches to the alveolar epithelium might facilitate subsequent influenza infection and may exacerbate influenza-mediated alveolar flooding that can lead to acute respiratory failure and death. PMID- 15121637 TI - H(2)O(2) inhibits alveolar epithelial wound repair in vitro by induction of apoptosis. AB - Reactive oxygen species (ROS) are released into the alveolar space and contribute to alveolar epithelial damage in patients with acute lung injury. However, the role of ROS in alveolar repair is not known. We studied the effect of ROS in our in vitro wound healing model using either human A549 alveolar epithelial cells or primary distal lung epithelial cells. We found that H(2)O(2) inhibited alveolar epithelial repair in a concentration-dependent manner. At similar concentrations, H(2)O(2) also induced apoptosis, an effect seen particularly at the edge of the wound, leading us to hypothesize that apoptosis contributes to H(2)O(2)-induced inhibition of wound repair. To learn the role of apoptosis, we blocked caspases with the pan-caspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp (zVAD). In the presence of H(2)O(2), zVAD inhibited apoptosis, particularly at the wound edge and, most importantly, maintained alveolar epithelial wound repair. In H(2)O(2) exposed cells, zVAD also maintained cell viability as judged by improved cell spreading and/or migration at the wound edge and by a more normal mitochondrial potential difference compared with cells not treated with zVAD. In conclusion, H(2)O(2) inhibits alveolar epithelial wound repair in large part by induction of apoptosis. Inhibition of apoptosis can maintain wound repair and cell viability in the face of ROS. Inhibiting apoptosis may be a promising new approach to improve repair of the alveolar epithelium in patients with acute lung injury. PMID- 15121636 TI - Cigarette smoke induces MUC5AC mucin overproduction via tumor necrosis factor alpha-converting enzyme in human airway epithelial (NCI-H292) cells. AB - Chronic obstructive pulmonary disease (COPD) is one of the leading causes of death in the U.S. Because cigarette smoking is so importantly implicated in the pathogenesis of COPD and because mucus hypersecretion plays such an important role in COPD, understanding of the mechanisms of smoking-induced mucus hypersecretion could lead to new therapies for COPD. Cigarette smoke causes mucin overproduction via EGF receptor (EGFR) in airway epithelial cells, but the cellular mechanism remains unknown. Airway epithelial cells contain EGFR proligands on their surfaces, which can be cleaved by metalloprotease and subsequently bind to EGFR resulting in mucin production. We hypothesize that TNF alpha-converting enzyme (TACE) is activated by cigarette smoke, resulting in increased shedding of EGFR proligand, leading to EGFR phosphorylation and mucin induction in human airway epithelial (NCI-H292) cells. Here we show that cigarette smoke increases MUC5AC production in NCI-H292 cells, an effect that is prevented by an EGFR-neutralizing antibody and by specific knockdown of transforming growth factor-alpha (TGF-alpha) using small interfering RNA (siRNA) for TGF-alpha, implicating TGF-alpha-dependent EGFR activation in the responses. Cigarette smoke increases TGF-alpha shedding, EGFR phosphorylation, and mucin production, which are prevented by metalloprotease inhibitors (GM-6001 and TNF alpha protease inhibitor-1) and by specific knockdown of TACE with TACE siRNA, implicating TACE in smoking-induced responses. Furthermore, pretreatment with antioxidants prevents smoking-induced TGF-alpha shedding and mucin production, suggesting that reactive oxygen species is involved in TACE activation. These results implicate TACE in smoking-induced mucin overproduction via the TACE proligand-EGFR signal pathway in NCI-H292 cells. PMID- 15121638 TI - 8-Bromo-cAMP decreases the Ca2+ sensitivity of airway smooth muscle contraction through a mechanism distinct from inhibition of Rho-kinase. AB - To clarify whether cyclic AMP (cAMP)/cAMP-dependent protein kinase (PKA) activation and Rho-kinase inhibition share a common mechanism to decrease the Ca2+ sensitivity of airway smooth muscle contraction, we examined the effects of 8-bromoadenosine 3',5'-cyclic monophosphate (8-BrcAMP), a stable cAMP analog, and (+)-(R)-trans-4-(1-aminoethyl)-N-(4-pyridyl) cyclohexane carboxamide dihydrochloride, monohydrate (Y-27632), a Rho-kinase inhibitor, on carbachol (CCh)-, guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS)-, 4beta-phorbol 12,13 dibutyrate (PDBu)-, and leukotriene D4 (LTD4)-induced Ca2+ sensitization in alpha toxin-permeabilized rabbit tracheal and human bronchial smooth muscle. In rabbit trachea, CCh-induced smooth muscle contraction was inhibited by 8-BrcAMP and Y 27632 to a similar extent. However, GTPgammaS-induced smooth muscle contraction was resistant to 8-BrcAMP. In the presence of a saturating concentration of Y 27632, PDBu-induced smooth muscle contraction was completely reversed by 8 BrcAMP. Conversely, PDBu-induced smooth muscle contraction was resistant to Y 27632. In the presence of a saturating concentration of 8-BrcAMP, GTPgammaS induced Ca2+ sensitization was also reversed by Y-27632. The 8-BrcAMP had no effect on the ATP-triggered contraction of tracheal smooth muscle that had been treated with calyculin A in rigor solutions. The 8-BrcAMP and Y-27632 additively accelerated the relaxation rate of PDBu- and GTPgammaS-treated smooth muscle under myosin light chain kinase-inhibited conditions. In human bronchus, LTD4 induced smooth muscle contraction was inhibited by both 8-BrcAMP and Y-27632. We conclude that cAMP/PKA-induced Ca2+ desensitization contains at least two mechanisms: 1) inhibition of the muscarinic receptor signaling upstream from Rho activation and 2) cAMP/PKA's preferential reversal of PKC-mediated Ca2+ sensitization in airway smooth muscle. PMID- 15121639 TI - Endotoxin responsiveness of human airway epithelia is limited by low expression of MD-2. AB - The expression of inducible antimicrobial peptides, such as human beta-defensin-2 (HBD-2) by epithelia, comprises a component of innate pulmonary defenses. We hypothesized that HBD-2 induction in airway epithelia is linked to pattern recognition receptors such as the Toll-like receptors (TLRs). We found that primary cultures of well-differentiated human airway epithelia express the mRNA for TLR-4, but little or no MD-2 mRNA, and display little HBD-2 expression in response to treatment with purified endotoxin +/- LPS binding protein (LBP) and soluble CD14. Expression of endogenous MD-2 by transduction of airway epithelial cells with an adenoviral vector encoding MD-2 or extracellular addition of recombinant MD-2 both increased the responses of airway epithelia to endotoxin + LBP and sCD14 by >100-fold, as measured by NF-kappaB-luciferase activity and HBD 2 mRNA expression. MD-2 mRNA could be induced in airway epithelia by exposure of these cells to specific bacterial or host products (e.g., killed Haemophilus influenzae, the P6 outer membrane protein from H. influenzae, or TNF-alpha + IFN gamma). These findings suggest that MD-2, either coexpressed with TLR-4 or secreted when produced in excess of TLR-4 from neighboring cells, is required for airway epithelia to respond sensitively to endotoxin. The regulation of MD-2 expression in airway epithelia and pulmonary macrophages may serve as a means to modify endotoxin responsiveness in the airway. PMID- 15121640 TI - Research on persons with impaired decision making and the public trust. PMID- 15121641 TI - Pharmacogenetics of psychotropic drug response. AB - OBJECTIVE: Molecular genetic approaches provide a novel method of dissecting the heterogeneity of psychotropic drug response. These pharmacogenetic strategies offer the prospect of identifying biological predictors of psychotropic drug response and could provide the means of determining the molecular substrates of drug efficacy and drug-induced adverse events. METHOD: The authors discuss methods issues in executing pharmacogenetic studies, review the first generation of pharmacogenetic studies of psychotropic drug response, and consider future directions for this rapidly evolving field. RESULTS: Pharmacogenetics has been most commonly used in studies of antipsychotic drug efficacy, antidepressant drug response, and drug-induced adverse effects. Data from antipsychotic drug studies indicate that polymorphisms within the serotonin 2A and dopamine receptor 2 genes may influence drug efficacy in schizophrenia. Moreover, a growing body of data suggests a relationship between the serotonin transporter gene and clinical effects of the selective serotonin reuptake inhibitors used to treat depression. A significant relationship between genetic variation in the cytochrome P450 system and drug-induced adverse effects may exist for certain medications. Finally, a number of independent studies point to a significant effect of a dopamine D(3) receptor polymorphism on susceptibility to tardive dyskinesia. CONCLUSIONS: Initial research into the pharmacogenetics of psychotropic drug response suggests that specific genes may influence phenotypes associated with psychotropic drug administration. These results remain preliminary and will require further replication and validation. New developments in molecular biology, human genomic information, statistical methods, and bioinformatics are ongoing and could pave the way for the next generation of pharmacogenetic studies in psychiatry. PMID- 15121643 TI - Pater familias. PMID- 15121642 TI - Proxy and surrogate consent in geriatric neuropsychiatric research: update and recommendations. AB - OBJECTIVE: The ethics of involving decisionally incapable adults in research will continue to grow in importance as more research is conducted to address the problems of decisionally impaired persons, especially elderly persons. The authors provide an updated discussion, critique, and recommendations regarding the need for clear legal and regulatory policy on this issue. METHOD: The authors summarize and build on discussions of a workshop on proxy and surrogate consent in geriatric neuropsychiatric research sponsored by the Aging Research Consortium of the National Institute of Mental Health. They incorporate the views of various stakeholders present at the workshop as well as review recent federal and state initiatives, recent empirical research and media reports, and various commission reports and relevant regulations. RESULTS: Despite a wave of initiatives in the late 1990s to clarify policy, surrogate consent for research continues to be a murky legal area and incapable subjects in the United States still lack clear regulatory protection. There is evidence that conservative risk management strategies by institutional review boards and their institutions may severely restrict research with decisionally impaired subjects. A passive approach to this problem may no longer be feasible. A recent federal advisory report on human research protections and legislative initiatives in some states could begin to provide a blueprint for future policy making. Interim recommendations for various stakeholders are given. CONCLUSIONS: It is imperative that the scientific community, patients and their advocates, and policy makers at all levels establish a constructive dialogue to clarify ethical and legal standards in the area of proxy and surrogate consent for research. PMID- 15121644 TI - Virginia Woolf (1882-1941). PMID- 15121645 TI - Randomized, controlled trial of the effectiveness of short-term dynamic psychotherapy and cognitive therapy for cluster C personality disorders. AB - OBJECTIVE: This study compared the effectiveness of short-term dynamic psychotherapy and cognitive therapy for outpatients with cluster C personality disorders. METHOD: Patients (N=50) who met the criteria for one or more cluster C personality disorders and not for any other personality disorders were randomly assigned to receive 40 weekly sessions of short-term dynamic psychotherapy or cognitive therapy. The most common axis I disorders in the patient group were anxiety and depression diagnoses. Therapists were experienced, full-time clinicians and were receiving manual-guided supervision. Outcome variables included symptom distress, interpersonal problems, and core personality pathology. Measures were administered repeatedly during and after treatment, and change was assessed longitudinally by means of growth modeling procedures. RESULTS: The overall patient group showed, on average, statistically significant improvements on all measures during treatment and also during a 2-year follow-up period. Significant changes in symptom distress after treatment were found for the group of patients who received short-term dynamic psychotherapy but not for the cognitive therapy patients. Despite these differences in intragroup changes, no statistically significant differences between the short-term dynamic psychotherapy group and cognitive therapy group were found on any measure for any time period. Two years after treatment, 54% of the short-term dynamic psychotherapy patients and 42% of the cognitive therapy patients had recovered symptomatically, whereas approximately 40% of the patients in both groups had recovered in terms of interpersonal problems and personality functioning. CONCLUSIONS: Both short-term dynamic psychotherapy and cognitive therapy have a place in the treatment of patients with cluster C personality disorders. However, factors other than treatment modality may discriminate better between successful and poor outcomes. Such factors should be explored in future studies. PMID- 15121646 TI - A PET study of dopamine D2 and serotonin 5-HT2 receptor occupancy in patients with schizophrenia treated with therapeutic doses of ziprasidone. AB - OBJECTIVE: Ziprasidone is an atypical antipsychotic drug that shows a higher affinity for serotonin 5-HT(2) receptors compared with dopamine D(2) receptors in vitro. The affinity of ziprasidone for these receptors in vivo in patients was examined in a positron emission tomography (PET) study. METHOD: The authors conducted a PET study to evaluate D(2) occupancy (using [(11)C]raclopride) and 5 HT(2) occupancy (using [(18)F]setoperone) in brain regions of interest in 16 patients with schizophrenia or schizoaffective disorder randomly assigned to receive 40, 80, 120, or 160 mg/day of ziprasidone, which reflected the recommended dose range. PET scanning was done after 3 weeks of administration and at trough plasma levels, i.e., 12-16 hours after the last dose. RESULTS: The mean 5-HT(2) receptor occupancy was significantly higher than the mean D(2) receptor occupancy (mean=76%, SD=15%, and mean=56%, SD=18%, respectively). The estimated plasma ziprasidone concentration associated with 50% maximal 5-HT(2) receptor occupancy was almost four times lower than that for D(2) receptor occupancy. CONCLUSIONS: These data affirm that ziprasidone is similar to other novel antipsychotics in having greater 5-HT(2) than D(2) receptor occupancy at therapeutic doses and suggest that the optimal effective dose of ziprasidone is closer to 120 mg/day than to the lower doses suggested by previous PET studies. The relatively high D(2) receptor occupancy, even at trough plasma levels, suggests that ziprasidone is more similar to risperidone and olanzapine in receptor occupancy profile than to clozapine and quetiapine. Since ziprasidone plasma levels show significant (more than twofold) variation within a single dose cycle, studies that are aimed at peak plasma levels (6 hours after the last dose) and that examine extrastriatal regions are required to fully characterize the in vivo occupancy profile of ziprasidone. PMID- 15121648 TI - A randomized, placebo-controlled trial of sertraline for prophylactic treatment of highly recurrent major depressive disorder. AB - OBJECTIVE: Previous antidepressant maintenance trials have used the same medication from acute through maintenance phases, confounding the interpretation of prophylactic effects. The purpose of this study was to determine whether sertraline prevents the recurrence of major depressive disorder among patients with recurrent depression who had been treated to remission with medications other than sertraline. METHOD: Patients who had experienced at least three documented episodes of major depressive disorder within the last 4 years and who were currently in full remission were eligible. The last episode must have been treated for at least 4 months with any antidepressant except sertraline. For the initial single-blind placebo lead-in phase, 371 patients were included; 288 were included in the analyses for the 18-month double-blind phase in which patients were randomly assigned to sertraline (50 or 100 mg) or placebo (two capsules per day). Recurrence was defined as a depressive episode that fulfilled DSM-IV criteria or the appearance of symptoms that required the administration of another antidepressant treatment. RESULTS: Sixty-one patients discontinued before the double-blind phase, including 33 who experienced a relapse. Out of the 288 who entered the double-blind prophylactic phase, 123 discontinued, including 65 for recurrences. Recurrences were significantly lower in the sertraline groups compared with placebo (sertraline, 50 mg: 16 [16.8%] of 95; sertraline, 100 mg: 16 [17.0%] of 94; placebo: 33 [33.3%] of 99). Patients treated with sertraline also had a significantly longer time until recurrence compared with placebo treated patients. CONCLUSIONS: Among remitted patients with a history of multiple depressive episodes, sertraline at a dose of either 50 or 100 mg/day prevented recurrences significantly more than did placebo. PMID- 15121647 TI - Serotonin transporter occupancy of five selective serotonin reuptake inhibitors at different doses: an [11C]DASB positron emission tomography study. AB - OBJECTIVE: Minimum therapeutic doses of paroxetine and citalopram produce 80% occupancy for the serotonin (5-HT) transporter (5-HTT). The authors used [(11)C]DASB positron emission tomography to measure occupancies of three other selective serotonin reuptake inhibitors (SSRIs) at minimum therapeutic doses. The relationship between dose and occupancy was also investigated. METHOD: Striatal 5 HTT binding potential was measured in 77 subjects before and after 4 weeks of medication administration. Binding potential is proportional to the density of receptors not blocked by medication. Subjects received citalopram, fluoxetine, sertraline, paroxetine, or extended-release venlafaxine. Healthy subjects received subtherapeutic doses; subjects with mood and anxiety disorders received therapeutic doses. Percent reduction in 5-HTT binding potential for each medication and dose was calculated. To obtain test-retest data, binding potential was measured before and after 4 weeks in six additional healthy subjects. RESULTS: Substantial occupancy occurred at subtherapeutic doses for all SSRIs. Compared to test-retest data, each drug at the minimum therapeutic dose had a significant effect on striatal 5-HTT binding potential. Mean occupancy at this dose was 76%-85%. At higher plasma SSRI concentrations, 5-HTT occupancy tended to increase above 80%. For each drug, as the dose (or plasma level) increased, occupancy increased nonlinearly, with a plateau for higher doses. CONCLUSIONS: At tolerable doses, SSRIs have increasing occupancy with increasing plasma concentration or dose. Occupancy of 80% across five SSRIs occurs at minimum therapeutic doses. This suggests that 80% 5-HTT blockade is important for therapeutic effect. Occupancy should be measured during development of antidepressant compounds targeting the 5-HTT. PMID- 15121649 TI - Neuropathological substrates of psychiatric symptoms in prospectively studied patients with autopsy-confirmed dementia with lewy bodies. AB - OBJECTIVE: This investigation was undertaken to clarify the neuropathological substrates of key psychiatric symptoms in dementia with Lewy bodies. METHOD: The authors studied 112 autopsy-confirmed cases of dementia with Lewy bodies in patients who had had annual standardized clinical evaluations until their death. The relationships of persistent psychiatric symptoms (visual hallucinations, delusions, depression) to plaques (Consortium to Establish a Registry for Alzheimer's Disease protocol), tangles (Braak staging), and Lewy bodies (consensus Lewy body staging) were evaluated. In addition, symptom frequency and persistent symptoms were compared in the patients with Lewy body dementia and 90 patients with autopsy-confirmed Alzheimer's disease studied prospectively during life. RESULTS: The main neuropathological correlate of persistent visual hallucinations was the presence of less severe tangle pathology, but there was no significant association between tangle pathology and persistent delusions. Lewy body staging was associated with the presence of persistent visual hallucinations and persistent delusions. All baseline psychiatric features were significantly more frequent in dementia with Lewy bodies than in Alzheimer's disease, as were persistent visual hallucinations, but patients who had dementia with Lewy bodies and severe tangle pathology had a clinical symptom profile more similar to that of Alzheimer's disease patients and were less likely to have neocortical Lewy bodies. CONCLUSIONS: The modest proportion of patients with Lewy body dementia and more severe tangle pathology resembled Alzheimer's disease patients clinically. Unlike Alzheimer's disease, dementia with Lewy bodies showed a significant inverse association between tangle burden and psychosis. PMID- 15121650 TI - Sustained benefit of supportive intervention for depressive symptoms in caregivers of patients with Alzheimer's disease. AB - OBJECTIVE: The long-term effect of counseling and support on symptoms of depression was examined in spouse-caregivers of patients with Alzheimer's disease. METHOD: The participants were 406 spouse-caregivers of Alzheimer's disease patients who lived at home at baseline. The caregivers were randomly assigned to either a group receiving enhanced counseling and support treatment or a group receiving usual care (control group). Caregivers in the enhanced treatment group were provided with six sessions of individual and family counseling, agreed to join support groups 4 months after enrollment, and received ongoing ad hoc counseling. The Geriatric Depression Scale was administered at baseline and at regular follow-up intervals for as long as the caregiver participated in the study. RESULTS: After baseline differences were controlled for, caregivers in the enhanced treatment group had significantly fewer depressive symptoms after the intervention than did the control subjects. These effects were sustained for 3.1 years after baseline, similar across gender and patient severity level, and sustained after nursing home placement or death of the patient. CONCLUSIONS: Counseling and support lead to sustained benefits in reducing depressive symptoms in spouse-caregivers of Alzheimer's disease patients and should be widely available to provide effective, evidence-based intervention for family caregivers. PMID- 15121651 TI - Extent and cost of informal caregiving for older Americans with symptoms of depression. AB - OBJECTIVE: The purpose of this study was to obtain nationally representative estimates of the additional time and cost associated with informal caregiving for older Americans with depressive symptoms. METHOD: Data from the 1993 Asset and Health Dynamics Among the Oldest Old Study, a nationally representative survey of people age 70 years or older (N=6,649), were used to determine the weekly hours and imputed costs of informal caregiving for elderly people with no depressive symptoms in the last week, one to three depressive symptoms in the last week, and four to eight depressive symptoms in the last week. RESULTS: Forty-four percent of survey respondents reported one to three depressive symptoms, and 18% reported four to eight depressive symptoms. In multivariate regression analyses that adjusted for sociodemographics, caregiver network, and coexisting chronic health conditions, respondents with no depressive symptoms received an average of 2.9 hours per week of informal care, compared with 4.3 hours per week for those with one to three symptoms and 6.0 hours per week for those with four to eight symptoms. Caregiving associated with depressive symptoms in elderly Americans represented a yearly cost of about $9 billion. CONCLUSIONS: Depressive symptoms in elderly persons are independently associated with significantly higher levels of informal caregiving, even after the effects of major coexisting chronic conditions are adjusted. The additional hours of care attributable to depressive symptoms represent a significant time commitment for family members and, therefore, a significant societal economic cost. Further research should evaluate the causal pathways by which depressive symptoms lead to high levels of caregiving and should examine whether successful treatment of depression reduces the need for informal care. PMID- 15121652 TI - Racial differences in paranoid ideation and psychoses in an older urban population. AB - OBJECTIVE: This study examined whether there are racial differences in the prevalence of paranoid ideation and psychotic symptoms in persons age >/=55 in an urban community. METHOD: Using 1990 census data for Brooklyn, N.Y., the authors attempted to interview all cognitively intact persons age >/=55 in randomly selected blocks. The final group consisted of 206 whites and 821 blacks. The authors used George's Social Antecedent Model for analyzing 21 independent and three dependent variables: paranoid ideation, psychotic symptoms, and psychotic symptoms/paranoid ideation. The group was weighted by race and gender. To control for intrablock clustering effects without replacement sampling, the authors used SUDAAN for data analysis. RESULTS: A significant difference in psychotic symptoms or paranoid ideation was found between blacks and whites (24% versus 10%) that was attenuated but not eliminated with logistic regression analyses. Blacks with psychotic symptoms or paranoid ideation, especially Caribbeans, had significantly lower receipt of mental health services and lower perceived service needs. With logistic regression, psychotic symptoms and paranoid ideation were associated with four variables among blacks and whites, although only one was significant in both groups. CONCLUSIONS: Racial differences in psychotic symptoms and paranoid ideation persist even after control for various clinical, social, and attitudinal effects. Among blacks, response to stressors may be expressed through increased paranoid ideation and psychotic symptoms. Stronger beliefs in spiritualism increase this expression in both races. The high prevalence of psychotic symptoms or paranoid ideation among this aging urban population, especially blacks, highlights a potential public health issue. PMID- 15121653 TI - Selective attention in schizophrenia: sparing and loss of executive control. AB - OBJECTIVE: Auditory selective attention involves top-down modulation of sensory processing to selectively filter relevant from irrelevant information. Patients with schizophrenia have deficits in auditory selective attention, but whether these attention deficits are evident at the earliest stages of auditory processing or emerge later in the processing stream is unknown. METHOD: Event related potentials were used to assess the integrity and time course of auditory attention in schizophrenia during a cross-modal selective attention task. A random sequence of equiprobable loud and soft speech sounds and bright and dim checkerboard patterns occurred every 800 to 1200 msec. A button press was required in response to soft speech sounds in the auditory attention task and to dim checkerboards in the visual attention task. Event-related potentials were recorded from 15 patients with DSM-IV schizophrenia and 16 age- and gender matched healthy comparison subjects. RESULTS: As early as 50 msec, both patients and comparison subjects showed evidence of cross-modal selective attention. These attention effects were sustained until 300 msec in comparison subjects but were no longer evident in patients by 100 msec. CONCLUSIONS: Schizophrenia patients implemented an early attentional filter but failed to sustain selective attention later in the processing stream. This suggests that initially intact executive control of auditory attention cannot be sustained in schizophrenia, perhaps implicating dysfunction in frontotemporal pathways. PMID- 15121654 TI - Glial cell loss in the anterior cingulate cortex, a subregion of the prefrontal cortex, in subjects with schizophrenia. AB - OBJECTIVE: Structural deficits in the anterior cingulate cortex such as changes in glial cell and neuron numbers may be part of the anatomical substrate for schizophrenia and need to be investigated. The total number of neurons and glial cells in brains of 12 schizophrenia subjects and 14 comparison subjects were determined in two subdivisions of the prefrontal cortex: Brodmann's area 24, a part of the anterior cingulate cortex, and Brodmann's area 32 in the paracingulate cortex. METHOD: The estimate of the total cell number was obtained by multiplying the volume of the region (estimated by using Cavalieri's point counting method) by the numerical density obtained from optical disectors in the cytoarchitectonically defined areas from the prefrontal cortex. RESULTS: The average total of bilateral glial cells in Brodmann's area 24 was 201 x 10(6 )in subjects with schizophrenia and 302 x 10(6 )in comparison subjects, a statistically significant difference of 33%, whereas there was a nonsignificant difference between the schizophrenia subjects and the comparison subjects in total number of glial cells in Brodmann's area 32. The bilateral average total number of neurons in areas 24 and 32 did not differ significantly between the schizophrenia and comparison subjects. CONCLUSIONS: A selective reduction in glial cells in Brodmann's area 24 (but not in area 32) is seen in brains of subjects with schizophrenia relative to those of comparison subjects. Further investigations of the glial cells, their mutual relationship, and their relationship with neurons are needed to understand the role of specific glial components in this mental disorder. PMID- 15121655 TI - Elevated maternal interleukin-8 levels and risk of schizophrenia in adult offspring. AB - OBJECTIVE: Many studies have implicated prenatal infection in the etiology of schizophrenia. Cytokines, a family of soluble polypeptides, are critically important in the immune response to infection and in other inflammatory processes. The goal of this study was to determine whether second-trimester levels of four cytokines-interleukin-8 (IL-8), interleukin-1beta (IL-1beta), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha)-are higher in the mothers of offspring who later developed schizophrenia spectrum disorders than in matched comparison subjects. METHOD: The authors conducted a nested case control study of maternal serum cytokine levels in a large birth cohort, born 1959-1967. Cases (N=59) were subjects diagnosed with schizophrenia spectrum disorders (mostly schizophrenia and schizoaffective disorder) who had available second-trimester maternal serum samples. Comparison subjects (N=105) were members of the birth cohort, had not been diagnosed with a schizophrenia spectrum disorder or major affective disorder, and were matched to subjects with schizophrenia for date of birth, gender, length of time in the cohort, and availability of maternal sera. Maternal second-trimester serum levels of IL-8, IL 1beta, IL-6, and TNF-alpha were determined by sandwich enzyme-linked immunosorbent assay. RESULTS: The second-trimester IL-8 levels in mothers of offspring with schizophrenia spectrum disorders were significantly higher than those of the mothers of comparison subjects. There were no differences between subjects with schizophrenia and comparison subjects with respect to maternal levels of IL-1beta, IL-6, or TNF-alpha. CONCLUSIONS: Using prospectively collected prenatal sera in a large and well-characterized birth cohort, the authors have documented a significant association between maternal IL-8 level during the second trimester and risk of schizophrenia spectrum disorders in the offspring. These findings provide further support for a substantive role of in utero infection or inflammation in the etiology of schizophrenia. Moreover, these results may have important implications for elucidating the mechanisms by which disrupted fetal development raises the risk of this disorder. PMID- 15121656 TI - Abnormalities of thalamic volume and shape in schizophrenia. AB - OBJECTIVE: Postmortem and neuroimaging studies of schizophrenia have reported deficits in the volume of the thalamus and its component nuclei. However, the pattern of shape change associated with such volume loss has not been investigated. In this study, alterations in thalamic volume, shape, and symmetry were compared in subjects with and without schizophrenia. METHOD: T(1)-weighted magnetic resonance scans were collected in 52 schizophrenia and 65 comparison subjects matched for age, gender, race, and parental socioeconomic status. High dimensional (large-deformation) brain mapping was used to assess thalamic morphology. RESULTS: Significant differences in thalamic volume, deformities of thalamic shape at the anterior and posterior extremes of the structure, and a significant exaggeration of thalamic asymmetry (i.e., left smaller than right) were found in the schizophrenia subjects. After covarying for total cerebral volume, the difference in thalamic volume became insignificant. When information about thalamic shape was combined with previously collected information about hippocampal shape, the discrimination between schizophrenia patients and comparison subjects was improved. CONCLUSIONS: Thalamic volume was smaller than normal in schizophrenia patients, but only proportionate to reductions in reduced total cerebral volume. The presence of changes in thalamic shape and asymmetry suggest greater pathologic involvement of individual nuclei at its anterior and posterior extremes of the thalamic complex. PMID- 15121657 TI - Risk for cancer in parents of patients with schizophrenia. AB - OBJECTIVE: This study attempted to determine whether a genetic protection against cancer might be manifest in parents of offspring with schizophrenia. METHOD: Using data from the Danish Central Population Registry, the authors identified 1,999,072 parents of offspring born after 1935. By linking this nationwide population-based parent cohort to the Danish Psychiatric Central Register, they identified 19,856 parents of offspring with schizophrenia. Follow-up for cancer in the Danish Cancer Registry began on the date of birth of the oldest child or April 1, 1969, and ended on the date of cancer diagnosis, death, or Dec. 31, 1997, yielding a total of 48,343,430 person-years at risk and 211,681 cases of cancer. The relative risk for cancer among parents with schizophrenic offspring compared to parents with no schizophrenic offspring was estimated by Poisson regression analysis and adjusted for age, period, and number of children. RESULTS: The risk for all cancer was 1.01 for fathers and 1.00 for mothers of schizophrenics. Mothers of schizophrenic patients had an increased risk of 1.20 for lung cancer and a nonsignificant risk of 1.14 for tobacco-related cancers combined. Apart from a reduced risk for leukemia in both mothers and fathers of schizophrenics, there was no difference in risk for any other cancer. DISCUSSION: This study does not confirm a previously reported reduced risk for cancer in parents of schizophrenic patients and provides no support for genetic protection against cancer in families with schizophrenia. PMID- 15121658 TI - Toward a rapidly acting antidepressant: the normetanephrine and extraneuronal monoamine transporter (uptake 2) hypothesis. AB - OBJECTIVE: The authors considered the possible role of the extraneuronal monoamine transporter (uptake 2) in accounting for the delay in clinical action of norepinephrine reuptake inhibitor antidepressant drugs. METHOD: Literature searches were performed by means of the MEDLINE and Current Contents databases with search terms such as "extraneuronal uptake," "uptake 2," "extraneuronal monoamine transporter," and "organic cation transporter type-3." RESULTS: The findings in this literature indicate that inhibition of glial uptake 2 by normetanephrine or other inhibitors of uptake 2 would enhance the accumulation of norepinephrine in the synapse. CONCLUSIONS: The authors propose the hypothesis that drugs or other agents that increase levels of normetanephrine or otherwise inhibit the extraneuronal monoamine transporter, uptake 2, in the brain will speed up the clinical effects of norepinephrine reuptake inhibitor antidepressant drugs. PMID- 15121659 TI - Depression and HIV risk behaviors among patients in a sexually transmitted disease clinic. AB - OBJECTIVE: The authors' goal was to investigate the relationship between depression and HIV risk behaviors or sexually transmitted disease (STD) diagnosis at an urban STD clinic. METHOD: Using audio computer-assisted self-interview, 671 STD clinic patients answered questions about HIV risk behaviors and depression in a large-scale, cross-sectional study. A subset of the patients (N=201) was evaluated for current major depressive disorder by interviewers using the nonpatient edition of the Structured Clinical Interview for DSM-IV. RESULTS: Depressed patients were more likely to have sex for money or drugs, to have had sex with an intravenous drug user, to have sex when "high" on alcohol or drugs, to have a greater number of lifetime sex partners, and to abuse alcohol or drugs than were nondepressed patients. Associations of HIV risk behaviors with depression persisted after adjustment for substance abuse. CONCLUSIONS: Depression in STD clinic patients is associated with HIV risk behaviors but not STD diagnosis. Identifying depression and developing strategies to intervene effectively may reduce HIV risk behaviors and improve health outcomes. PMID- 15121660 TI - Early-stage face processing dysfunction in patients with schizophrenia. AB - OBJECTIVE: Deficits in emotional processing are evident in patients with schizophrenia. Since recent studies have indicated dysfunctions in cortical areas involved in face processing, this study investigated the early processing of faces in schizophrenia patients through the use of event-related potentials. METHOD: Event-related potential responses were obtained from 24 schizophrenia patients and 28 healthy comparison subjects who were presented with pictures of faces and pictures of buildings (control stimuli). RESULTS: Schizophrenia patients displayed significantly lower differences in the face-specific N170 component between the face and building pictures than did the healthy comparison subjects. CONCLUSIONS: This finding suggests a dysfunction of early-stage visual processing of faces in patients with schizophrenia. PMID- 15121662 TI - Obstetric adversity and age at first presentation with schizophrenia: evidence of a dose-response relationship. AB - OBJECTIVE: The purpose of the study was to determine if a dose-response relationship exists between obstetric adversity and age at first presentation with schizophrenia. METHOD: The Dublin Psychiatric Case Register was used to identify subjects with schizophrenia. Data on obstetric complications, social class of origin, and family history of psychiatric illness were obtained for those subjects. RESULTS: A total of 409 patients with ICD-9 schizophrenia were identified. Patients with a history of obstetric complications presented earlier to psychiatric services. As the number of complications increased, the mean age at first presentation decreased. This effect was independent of social class of origin and family history of psychiatric illness. CONCLUSIONS: Obstetric adversity exerts an independent influence on the age at first presentation with schizophrenia, in a dose-response manner. This finding supports the existence of a causal relationship between obstetric adversity and age at first presentation with schizophrenia. PMID- 15121661 TI - Growth and sexual maturation during long-term treatment with risperidone. AB - OBJECTIVE: This study assessed the impact of risperidone on growth and sexual maturation. METHOD: The pooled database of five studies included 700 children ages 5-15 years with disruptive behavior disorders. All evaluable patients had received risperidone for 11 or 12 months. Those evaluable for growth also had baseline and 11- or 12-month height measurements (N=350); girls >/=9 years and boys >/=10 years who were evaluable for sexual maturation also had baseline and 11- or 12-month Tanner staging (N=222). RESULTS: Risperidone-treated children had a mean increase in height 1.2 cm greater than the reference population, and they experienced no delay in progression through Tanner staging. Transient increases in prolactin did not correlate with growth or sexual maturation. CONCLUSIONS: In this retrospective analysis, there was no evidence of statistically or clinically significant growth failure or delay in pubertal onset or progression in children treated for up to 1 year with risperidone. PMID- 15121664 TI - Characteristics of functional auditory hallucinations. PMID- 15121663 TI - Hepatotoxicity related to citalopram. PMID- 15121665 TI - Clozapine with amisulpride for refractory schizophrenia. PMID- 15121666 TI - Is multiple cavernoma a developmental defect in schizophrenia? PMID- 15121667 TI - Antidepressants and premature labor. PMID- 15121669 TI - Diagnostic stability of personality disorders. PMID- 15121670 TI - Body dysmorphic disorder and art background. PMID- 15121673 TI - Transcranial magnetic stimulation and processing of facial threats. PMID- 15121675 TI - Depression severity and HIV risk behavior. PMID- 15121676 TI - Checklist psychiatry's effect on psychiatric education. PMID- 15121677 TI - Name for new class of antipsychotic medications. PMID- 15121679 TI - Violent behavior among Colombian adolescents. PMID- 15121683 TI - Ethnic differences in ADHD and the mad/bad debate. PMID- 15121684 TI - Autism and difficulty levels in social visual pursuit. PMID- 15121686 TI - Comorbidity of gender identity disorders. PMID- 15121687 TI - Comorbidity of gender identity disorders. PMID- 15121698 TI - A uniform approach to setting and using DRLs? PMID- 15121699 TI - Whole-body CT health screening. PMID- 15121700 TI - Comparison of new clinical and scintigraphic algorithms for the diagnosis of pulmonary embolism. AB - Since the publication of the modified Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) criteria for the diagnosis of pulmonary embolism (PE), new clinical and scintigraphic diagnostic algorithms (the McMaster clinical criteria, the PisaPED simplified scintigraphic grading and the Miettinen logistic regression analysis) have been reported although the results have not been reproduced in other sites. Ventilation-perfusion lung scintigraphy was performed in 238 consecutive patients with a provisional diagnosis of PE. Scans were reported as normal/very low, low, intermediate or high probability for PE using standardized criteria. Each patient received a clinical grading of probability of PE as low, moderate or high using the McMaster clinical criteria. Using the PisaPED criteria (an alternate simplified scintigraphic grading system using the perfusion scan alone) each scan was also graded as normal/near normal, abnormal but not PE, or abnormal and PE. Using the logistic regression algorithm of Miettinen each scan received a numerical probability of PE. Frequencies for differing levels of probability of PE varied widely between the various algorithms. Cross tabulations revealed correlation of the standardized criteria with the Miettinen grading but not with the McMaster or the PisaPED gradings. We were unable to reproduce similar results using the McMaster clinical grading or the PisaPED simplified scintigraphic grading although the Miettinen logistic regression formula gave comparable results. New algorithms are not automatically transferable to new environments. PMID- 15121701 TI - Development of a simultaneous boost IMRT class solution for a hypofractionated prostate cancer protocol. AB - The purpose of this work was to develop a robust technique for planning intensity modulated radiation therapy (IMRT) for prostate cancer patients who are to be entered into a proposed hypofractionated dose escalation study. In this study the dose escalation will be restricted to the prostate alone, which may be regarded as a concurrent boost volume within the overall planning target volume (PTV). The dose to the prostate itself is to be delivered in 3 Gy fractions, and for this phase of the study the total prostate dose will be 57 Gy in 19 fractions, with 50 Gy prescribed to the rest of the PTV. If acute toxicity results are acceptable, the next phase will escalate doses to 60 Gy in 20 x 3 Gy fractions. There will be 30 patients in each arm. This work describes the class solution which was developed to create IMRT plans for this study, and which enabled the same set of inverse planning parameters to be used during optimization for every patient with minimal planner intervention. The resulting dose distributions were compared with those that would be achieved from a 3D conformal radiotherapy (3DCRT) technique that used a multileaf collimator (MLC) but no intensity modulation to treat the PTV, followed by a sequential boost to raise the prostate to 57 Gy. The two methods were tested on anatomical data sets for a series of 10 patients who would have been eligible for this study, and the techniques were compared in terms of doses to the target volumes and the organs at risk. The IMRT method resulted in much greater sparing of the rectum and bladder than the 3DCRT technique, whilst still delivering acceptable doses to the target volumes. In particular, the volume of rectum receiving the minimum PTV dose of 47.5 Gy was reduced from a mean value of 36.9% (range 23.4% to 61.0%) to 18.6% (10.3% to 29.0%). In conclusion, it was found possible to use a class solution approach to produce IMRT dose escalated plans. This IMRT technique has since been implemented clinically for patients enrolled in the hypofractionated dose escalation study. PMID- 15121702 TI - Infectious meningitis: prospective evaluation with magnetization transfer MRI. AB - The study was performed with the aim of prospectively characterizing infectious meningitis of different aetiology using magnetization transfer (MT) MRI. Spin echo (SE) T(1), T(2) and pre- and post-contrast T(1) weighted MT images in 100 patients with aetiologically proven meningitis were evaluated for the visibility and enhancement of the meninges on pre- and post-contrast T(1) weighted MT images, respectively. The MT ratio (MTR) was calculated from the thickened meninges in tuberculous meningitis. In addition, the percentage difference in the mean signal intensity (SI) of the meninges and adjacent brain parenchyma was calculated and compared between different groups using 2-tailed student's t-test. T(1) weighted MT images were highly sensitive (96%) in the detection of abnormal meningeal enhancement. Meninges were visible on pre-contrast T(1) weighted MT images only in patients with tuberculous meningitis. The MTR from meninges in tuberculous infection was 19.10+/-1.02, and the percentage difference in the mean SI of the meninges and the adjacent T(2) normal brain parenchyma was significantly higher (p<0.05) in the tuberculous group compared with that in the non-tuberculous group. MT MRI is an important technique for the detection and characterization of infectious meningitis of different aetiology. Visibility of the meninges on pre-contrast T(1) weighted MT images may be considered highly suggestive of tuberculous meningitis. PMID- 15121703 TI - Comprehensive analysis of the spectrometric determination of voltage applied to X ray tubes in the radiography and mammography energy ranges using a silicon PIN photodiode. AB - This work describes the analysis of factors which affect the results of estimation of the electron accelerating potential (kVp) applied to an X-ray tube, through determination of the end point of the energy spectrum of the emitted radiation beam. Measurements have been performed utilizing two spectrometers each with a silicon PIN photodiode: one operating at room temperature, and the other, a high resolution spectrometer, with a Peltier cooler. Both were directly irradiated by different X-ray beams. Both systems work at low voltage and without liquid nitrogen cooling, thus avoiding the drawbacks presented by germanium detectors. Each kVp value was determined by linear regression of the end of the spectrum, so as to give, simultaneously, the best fit to the experimental data and low standard deviation for the kVp value. Detector energy resolution and calibration, counting statistics and high voltage waveform ripple have been investigated in order to establish better experimental conditions and to optimize measurement time. Results of measurements carried out with X-ray tubes connected to single-phase, three-phase or constant potential units, using additional filtration of Cu, Al or Mo (for mammographic beams), are presented. The variations resulted in kVp uncertainties up to 0.1 kV. PMID- 15121705 TI - Comparison of a lung fitting algorithm with CT data for tangential fields in radiotherapy of the breast. AB - A method of estimating the shape and position of the lung in tangential breast fields is presented for patients who have not been CT scanned. Using the Osiris system, the external contour is obtained optically, and an estimated lung structure superimposed on the transverse outlines based on the measured lung depth in the tangential fields and an analysis of the typical lung shapes obtained from CT images. The accuracy of this fit was determined by comparison with a set of 64 CT images imported into the Osiris system. Dose distributions were calculated by two treatment planning systems: ADAC Pinnacle and GE Target2. The computed dose distributions for 6 MV photons were compared against measured doses in a specialized breast phantom. For the worst case of lung fit compared with CT, the dosimetric error (based upon ADAC Pinnacle calculations) was 2.0% in the shadow of the lung. For the complete patient data set, the relative dose errors to these points were reduced from a mean value of 8.4% and standard deviation (SD)=1.8% (no lung correction) to a mean of 0.2% and SD=1.0% (lung correction using fitted lung). It was also found that for every 1 cm of lung path length the dose to the breast along that path length increased by approximately 1%. The results of these investigations indicated that the lung fit model was satisfactory for routine clinical use, so that good dosimetric results can be obtained using lung correction without the need for CT imaging. PMID- 15121704 TI - Quantification of tumour response to radiotherapy. AB - In 1979, the World Health Organization (WHO) established criteria based on tumour volume change for classifying response to therapy as (i) progressive disease (PD), (ii) partial recovery (PR), and (iii) no change (NC). Typically, the tumour volume is reported from diameter measurements, using the calliper method. Alternatively, the Cavalieri method provides unbiased volume estimates of any structure without assumptions about its shape. In this study, we applied the Cavalieri method in combination with point counting to investigate the changes in tumour volume in four patients with high grade glioma, using 3D MRI. In particular, the volume of tumour within the enhancement boundary, the enhancing abnormality (EA), was estimated from T(1) weighted images, and the volume of the non-enhancing abnormality, (NEA) enhancing abnormality, was estimated from T(2) relaxation time and magnetic transfer ratio tissue characterization maps. We compared changes in tumour volume estimated by the Cavalieri method with those obtained using the calliper method. Absolute tumour volume differed significantly between the two methods. Analysis of relative change in tumour volume, based on the WHO criteria, provided a different classification using the calliper and Cavalieri methods. The benefit of the Cavalieri method over the calliper method in the estimation of tumour volume is justified by the following factors. First, Cavalieri volume estimates are mathematically unbiased. Second, the Cavalieri method is highly efficient under an appropriate sampling density (i.e. EA volume estimates can be obtained with a coefficient of error no higher than 5% in 2-3 min). Third, the source of variation of the volume estimates due to disagreements between observers, and within observer, is much greater in the positioning of the calliper diameters than in the identification of the tumour boundaries when applying the Cavalieri method. Additionally, the error prediction formula, available to estimate the coefficient of error of Cavalieri volume estimates from the data, allows us to establish more precise classification criteria against which to identify potentially clinical significant changes in tumour volume. PMID- 15121706 TI - X-ray imaging for palaeontology. AB - Few may be aware that X-ray imaging is used in palaeontology and has been used since as early as 1896. The X-raying, preparation and exposure of Hunsruck slate fossils are described. Hospital X-ray machines are used by the author in his work. An X-ray is vital to provide evidence that preparation of a slate is worthwhile as well as to facilitate preparation even if there is little external sign of what lies within. The beauty of the X-ray exposure is an added bonus. PMID- 15121707 TI - Towards in vivo TLD dosimetry in mammography. AB - While phantoms are used for quality control assessment of the mammography unit, in vivo dose measurements are necessary to account for the variation in size and composition of the female breast. The use of thermoluminescent dosimeters (TLDs) in mammography has been limited due to TLD visibility. The aim of this current investigation was to access the suitability of a paper-thin LiF:Mg,Cu,P TLD (GR 200F) for in vivo dosimetric mammography measurements. The visibility of GR-200F has been directly compared with LiF:Mg,Cu,P TLDs (GR-200A) using a number of commercially available phantoms. The phantoms of thickness 2-5 cm were imaged over the range of tube potentials (24-28 kVp) used clinically. Both types of TLD were placed on the surface of the phantoms allowing assessment of visibility, entrance surface dose (ESD) and field homogeneity. In vivo assessment of ESD and visibility was also carried out on a volunteer undergoing a routine mammography examination. The positions of the GR-200F TLDs were not identified either on the image of the Leeds TOR(MAM) phantom or the patient mammograms. The average ESD for the Leeds phantom was 8.8 mGy, while the patient ESD was 13 mGy. It is now possible to perform in vivo measurements with the potential of increasing the accuracy of the doses measured for women that do not conform to a standard breast thickness or density. PMID- 15121708 TI - Paracardiac pleural Castleman disease: radiographic and MR findings. AB - Castleman disease is a rare benign lymphoid tumour of uncertain aetiology that usually appears as a solitary mediastinal mass. We report a rare case of Castleman disease in the right paracardiac pleural space, occurring in a young woman with non-specific chest discomfort. MRI showed a well-defined, oval mass that was slightly hyperintense on T(1) weighted images, inhomogeneously hyperintense on T(2) and enhanced T(1) weighted images. The patient underwent radical tumour resection and has remained well for 8 years. PMID- 15121709 TI - Cellular congenital mesoblastic nephroma with contralateral medullary nephrocalcinosis. AB - Congenital mesoblastic nephroma is the most common renal mass in the newborn period and can present with atypical findings. Certain associated conditions such as hypercalcaemia, hypertension and reninism have been described. We report a cellular variant of congenital mesoblastic nephroma with hypercalcaemia and contralateral medullary nephrocalcinosis. PMID- 15121710 TI - Concomitant pulmonary and mediastinal sclerosing haemangiomas. AB - We report a case of thoracic sclerosing haemangioma with concomitant pulmonary and mediastinal involvement in a 19-year-old girl who presented with haemoptysis and dyspnoea. CT showed a large oval mass in the left lower lobe and another larger dumbbell-shaped mass in the posterior mediastinum. Both masses were well defined and harboured punctate calcifications. They exhibited inhomogeneous contrast enhancement and contained some cystic areas. In addition, an air meniscus sign was present around the pulmonary lesion. PMID- 15121711 TI - Coronary subclavian steal syndrome: non-invasive imaging and percutaneous repair. AB - Although coronary subclavian steal syndrome (CSSS) is relatively uncommon, it is a well documented cause of graft failure in patients having undergone coronary artery bypass grafting (CABG) using the left internal mammary artery (LIMA). Here we report a case of CSSS induced by restenosis of a left subclavian artery (SCA) origin stent, identified by increased velocities within the stent and an abnormal ipsilateral vertebral artery (VA) waveform on Duplex ultrasound imaging. This was successfully treated percutaneously by re-stenting, resulting in restoration of normal SCA waveforms and velocities, and normalization of the ipsilateral VA waveform. PMID- 15121712 TI - Small pulmonary arteriovenous fistulae revealed by scintigraphy during selective injection of 99Tc(m)-macroaggregated albumin. AB - We report a case of small pulmonary arteriovenous fistulae precisely diagnosed by a combination of selective pulmonary arteriography and scintigraphy during (99)Tc(m)-macroaggregated albumin injection via the peripheral pulmonary artery. PMID- 15121713 TI - Chest wall tuberculosis: a review of CT appearances. AB - Tuberculous abscesses of the chest wall, though uncommon are not infrequently encountered in countries endemic to the disease. This pictorial review of 14 patients highlights the varied appearance of tuberculosis (TB) of the chest wall on CT. The patients ranged in age from 9 to 55 years (a mean of 25 years) with a preponderance of chest wall lesions in young adults and in females (male to female ratio of 2:5). Cases in which there was no involvement of the chest wall other than of the spine have been excluded. In all cases CT demonstrated peripherally enhancing chest wall collections some of which were accompanied by changes in adjacent bone. Enlargement of intrathoracic lymph nodes with comparatively lesser involvement of lung parenchyma and pleura was also seen. PMID- 15121714 TI - A woman with a pelvic mass and pulmonary nodules. PMID- 15121715 TI - The defence of dirt. PMID- 15121716 TI - Cohort study of sibling effect, infectious diseases, and risk of atopic dermatitis during first 18 months of life. AB - OBJECTIVES: To determine whether early infectious diseases could explain the association between number of siblings and other markers of microbial exposure and the development of atopic dermatitis before the age of 18 months. DESIGN: Cohort study. Information on atopic dermatitis, infectious diseases occurring before 6 months of age, number of siblings, early day care, pet keeping, farm residence, and background factors was collected in telephone interviews. SETTING: Danish national birth cohort. PARTICIPANTS: 24,341 mother-child pairs. MAIN OUTCOME MEASURES: Incidence rate ratios of atopic dermatitis. RESULTS: 13,070 children (54%) had at least one clinically apparent infectious disease before 6 months of age. At age 18 months, 2638 (10.8%) of the children had had atopic dermatitis. The risk of atopic dermatitis increased with each infectious disease before 6 months of age (incidence rate ratio 1.08, 95% confidence interval 1.04 to 1.13). The risk of atopic dermatitis decreased with each additional exposure to three or more siblings, day care, pet ownership, and farm residence (0.86, 0.81 to 0.93). CONCLUSIONS: Early infections do not seem to protect against allergic diseases. The protective effect of number of siblings, day care, pet ownership, and farm residence remained after adjustment for clinically apparent infectious diseases, suggesting that the effect is established independently early in life. PMID- 15121717 TI - Apparent modulation of CD20 by rituximab: an alternative explanation. PMID- 15121718 TI - Severe hypochromic microcytic anemia caused by a congenital defect of the iron transport pathway in erythroid cells. PMID- 15121719 TI - S-Adenosyl-L-methionine-dependent restriction enzymes. AB - Restriction-modification (R-M) enzymes are classified into type I, II, III, and IV, based on their recognition sequence, subunit composition, cleavage position, and cofactor requirements. While the role of S-Adenosyl-L-methionine (AdoMet) as the methyl group donor in the methylation reaction is undisputed, its requirement in DNA cleavage reaction has been subject to intense study. AdoMet is a prerequisite for the DNA cleavage by most type I enzymes known so far, with the exception of R.EcoR124I. A number of new type II restriction enzymes belonging to the type IIB and IIG family were found to show AdoMet dependence for their cleavage reaction. The type III enzymes have been found to require AdoMet for their restriction function. AdoMet functions as an allosteric effector of the DNA cleavage reaction and has been shown to bring about conformational changes in the protein upon binding. PMID- 15121720 TI - FeII/alpha-ketoglutarate-dependent hydroxylases and related enzymes. AB - FeII/alpha-ketoglutarate (alphaKG)-dependent hydroxylases catalyze an amazing diversity of reactions that result in protein side-chain modifications, repair of alkylated DNA/RNA, biosynthesis of antibiotics and plant products, metabolism related to lipids, and biodegradation of a variety of compounds. These enzymes possess a beta-strand "jellyroll" structural fold that contains three metal binding ligands found in a His1-X-Asp/Glu-Xn-His2 motif. The cosubstrate, alphaKG, chelates FeII using its C-2 keto group (binding opposite the Asp/Glu residue) and C-1 carboxylate (coordinating opposite either His1 or His2). Oxidative decomposition of alphaKG forms CO2 plus succinate and leads to the generation of an FeIV-oxo or other activated oxygen species that hydroxylate the primary substrate. The reactive oxygen species displays alternate reactivity in related enzymes that catalyze desaturations, ring expansions, or ring closures. Other enzymes resemble the FeII/alphaKG-dependent hydroxylases in terms of protein structure or chemical mechanism but do not utilize alphaKG as a substrate. This review describes the reactions catalyzed by this superfamily of enzymes, highlights key active site features revealed by structural studies, and summarizes results from spectroscopic and other approaches that provide insights into the chemical mechanisms. PMID- 15121723 TI - Difficulty in advancing a tracheal tube over a fibreoptic bronchoscope: incidence, causes and solutions. PMID- 15121722 TI - NICE guidelines for central venous catheterization in children. Is the evidence base sufficient? AB - BACKGROUND: Recent guidelines from the National Institute for Clinical Excellence (NICE) recommend the use of ultrasound guidance for central venous catheterization in children. This study prospectively examined the use of ultrasound guidance for central venous catheterization in children undergoing heart surgery. METHODS: One hundred and twenty-four infants and children were randomized to either ultrasound-guided or traditional landmark-guided central venous catheterization. RESULTS: Success rates were significantly greater in the landmark group compared with the ultrasound group (89.3% vs 78%, P<0.002), and arterial puncture rates were significantly lower in the landmark group (6.2% vs 11.9%, P<0.03). There was no significant difference between the two groups in the time taken to perform the catheterization. CONCLUSIONS: These results are different from the published results on which the NICE guidelines were based; however, the evidence base in children is small. There is currently insufficient evidence to support the use of ultrasound guidance for central venous catheterization in children. PMID- 15121724 TI - Evaluation of simple criteria to predict successful weaning from mechanical ventilation in intensive care patients. AB - BACKGROUND: There is increasing evidence that weaning protocols improve outcome from mechanical ventilation, but it is unclear how best to implement such protocols in large intensive care units. We evaluated a checklist of simple bedside criteria to determine whether it could be used reliably to predict successful discontinuation of mechanical ventilation. METHODS: We carried out a prospective observational cohort study in a 12-bedded general intensive care unit (ICU). We developed a checklist of metabolic, cardiorespiratory and neurological criteria that suggested that patients should start the weaning process. We performed daily assessments throughout ICU stay and recorded whether the criteria were met. Ultimate ventilator independence was used as the reference standard. RESULTS: We studied 325 sequential admissions to the ICU. Data were available for 98% of patients; 97% of admissions were mechanically ventilated on admission to ICU. Overall, 205 of the 308 ventilated patients (67%) achieved ventilator independence during ICU admission; the other patients died or were transferred ventilated to other ICUs. Eighty-three per cent of the patients who achieved ventilator independence met the set criteria. Fulfilling the criteria was a moderately strong predictor of ultimate ventilator independence: specificity 89%, positive predictive value 94%, positive likelihood ratio (LR) 7.6. When we analysed data by the day from admission on which patients were examined, the test was a strong predictor of subsequent ventilator independence when criteria were met by day 1 (LR 11.1) or day 2 (LR 6.9), but weaker when met by more than/equal to 4 days (LR <3). Patients who met criteria after more than/equal to 4 days often had prolonged weaning and a high incidence of re-intubation. Patients who achieved ventilator independence without fulfilling the criteria (n=35) had a short duration of mechanical ventilation (median 2 days, interquartile range 1-3 days). The most frequent reason for failing criteria before ventilator independence was a Pa(O(2))/FI(O(2)) ratio less than 24 kPa (49% of cases). CONCLUSIONS: A simple checklist can assist nurse assessment of suitability for weaning and could be used as a trigger to commence a weaning protocol. The day on which criteria are met is a useful way of stratifying patients for likely patterns of weaning. PMID- 15121725 TI - Retrieval of a retrograde catheter using suction, in patients who cannot open their mouths. AB - In difficulty, tracheal intubation can be facilitated by passing a retrograde catheter, but the mouth has to be opened for the retrieval of the catheter from the pharynx. Two patients with ankylosis of a temporomandibular joint were unable to open their mouth, and required general anaesthesia for gap arthroplasty. Because we did not have a flexible fibreoptic laryngoscope, we used a suction catheter to retrieve an epidural catheter from the pharyngeal cavity, which had been passed retrogradely from a cricothyroid puncture. Catheter-guided tracheal intubation was done without complication. A suction catheter can assist retrograde retrieval of a catheter to aid intubation in patients who cannot open the mouth. PMID- 15121726 TI - Medical aerosol propellant interference with infrared anaesthetic gas monitors. AB - BACKGROUND: 1,1,1,2 Tetrafluoroethane is a hydrofluoroalkane (HFA) that is replacing chlorofluorocarbons (CFC) as a medical aerosol propellant in an attempt to reduce damage to the ozone layer. This study compared the effects of HFA- and CFC-based inhalers on four anaesthetic gas monitoring systems. METHODS: The HFA- and CFC-based inhalers were activated in close proximity to the sample line of two Datex Ohmeda, an Agilent and a Siemens infrared anaesthetic agent monitoring systems. The effects were recorded on each system for five common anaesthetic agents. RESULTS: The HFA inhaler caused either maximal false positive readings (with the exception of desflurane) or transient measurement failure on all systems. The Datex Ohmeda AS/3 system misidentified the HFA inhaler as carbon dioxide at low concentration (2 +/- 0 mm Hg). The CFC-based inhaler caused a minor false-positive reading (0.4 +/- 0%) for halothane only on the Datex Ohmeda AS/3 system only and was misidentified as carbon dioxide at 33.3 (sd 2.1) mm x Hg and 22.4 (8.9) mm x Hg by the Agilent and Siemens systems. CONCLUSIONS: The HFA inhaler adversely affected all equipment tested. The infrared spectra of HFA and the common anaesthetic gases have considerable overlap at the 8-12 microm range that is not shared by the CFCs. The differences in spectral overlap explain the different effects of the HFA and CFC propellants. Anaesthetic gas concentration data may be erroneous using the HFA-based inhalers. PMID- 15121727 TI - Early analgesic effects of parecoxib versus ketorolac following laparoscopic sterilization: a randomized controlled trial. AB - BACKGROUND: The aim of this prospective double blind randomized controlled trial was to compare the effects of ketorolac and parecoxib on early postoperative pain. METHOD: We studied 36 ASA I/II patients who received a standardized general anaesthetic for laparoscopic sterilization. Patients were allocated randomly to receive either parecoxib 40 mg i.v. or ketorolac 30 mg i.v., at induction. After surgery, patients were assessed on awakening and then at 1, 2, and 3 h. Abdominal pain at rest and on inspiration, in addition to nausea and sedation were assessed on a 100 mm visual analogue scale. RESULTS: Of 36 patients, one was excluded from analysis. In the remaining patients, pain scores at rest and on inspiration were significantly lower in patients given ketorolac compared with those given parecoxib. This difference was attributable to the higher pain scores on awakening and at 1 h postoperatively in the parecoxib group compared with the ketorolac group. Despite this initial difference, there was no significant difference between the two groups in the number of patients receiving rescue analgesia. The median (interquartile range) time to consumption of rescue cocodamol of 60 (46-74) min in the parecoxib group was not significantly shorter than that of 100 (70-130) min in the ketorolac group. The amount of cyclizine given, nausea and sedation did not differ significantly between the groups. CONCLUSION: We found that parecoxib 40 mg i.v. given at induction of anaesthesia was less effective than or ketorolac 30 mg i.v., in the first hour after laparoscopic sterilization. PMID- 15121729 TI - Preoperative optimization of the high-risk surgical patient. PMID- 15121728 TI - Physiology of the endothelium. AB - In the past, the endothelium was considered to be inert, described as a 'layer of nucleated cellophane', with only non-reactive barrier properties, such as presentation of a non-thrombogenic surface for blood flow and guarding against pro-inflammatory insults. However, it is now becoming clear that endothelial cells actively and reactively participate in haemostasis and immune and inflammatory reactions. They regulate vascular tone via production of nitric oxide, endothelin and prostaglandins and are involved in the manifestations of atherogenesis, autoimmune diseases and infectious processes. They produce and react to various cytokines and adhesion molecules and it is now clear that they can mount anti- and pro-inflammatory and protective responses depending on environmental conditions and are key immunoreactive cells. Endothelial dysfunction or activation also contributes to a variety of disease states. PMID- 15121730 TI - The heart and circulation in severe sepsis. PMID- 15121731 TI - Brief communication: birth month influences reproductive performance in contemporary women. AB - BACKGROUND: Season of birth has been reported to affect later reproduction in samples of pre-modern women and contemporary men. METHODS: To examine whether the effect of birth date is also valid in contemporary women, we investigated the association between birth month and measures of reproductive performance (number of live-born children, % childless individuals) in a representative sample of contemporary Austrian women. RESULTS: Among reproducing women, birth month is significantly associated with the number of live-born children (n = 2839, P = 0.032). On average, women born in summer months have fewer children than women born during the remainder of the year. No association between birth month and the percentage of childless individuals was found. CONCLUSIONS: As has been reported in pre-modern women, month of birth also appears to affect later reproduction in contemporary women. PMID- 15121732 TI - Demonstration of Chlamydia trachomatis IgG antibodies in the male partner of the infertile couple is correlated with a reduced likelihood of achieving pregnancy. AB - BACKGROUND: The objective of this study was to determine the prevalence of Chlamydia trachomatis among both men and women seeking help at an infertility clinic, and to prospectively follow the effect of previous infection on pregnancy rates and pregnancy outcome after a long follow-up period (mean 37 months). METHODS: A total of 244 infertile couples was tested for C. trachomatis IgG antibodies, and IgG(+) couples were also tested for C. trachomatis DNA by PCR in a first-void urine sample. Study parameters were serology, PCR results, clinical diagnoses, treatments, pregnancy rates and pregnancy outcome. As controls, age matched and spontaneously pregnant women were also tested with serology. RESULTS: The prevalence of IgG antibodies was 24.2, 20.1 and 15.6% among infertile women, infertile men and control women respectively. The prevalence of C. trachomatis DNA was 6.8 and 7.1% among tested women and men respectively. The presence of C. trachomatis IgG antibodies in women was related to tubal factor infertility (TFI) (P = 0.002). Decreased pregnancy rates were seen in couples where the man was IgG(+) (P = 0.005) with no relationship to TFI. Among women who achieved pregnancy, there was no difference in pregnancy outcome between IgG(+) or negative couples. CONCLUSIONS: C. trachomatis IgG antibodies in the man of the infertile couple was related to decreased pregnancy rates and to the presence of IgG antibodies in the woman. There was a high prevalence of asymptomatic persistent infections among infertile couples. PMID- 15121733 TI - Prospective observational study of bone mineral density during pregnancy: low molecular weight heparin versus control. AB - BACKGROUND: The aim of this study was to assess the effect of long-term low molecular weight heparin (LMWH) on bone mineral density (BMD) during pregnancy. METHODS: Fifty-five patients with recurrent miscarriage and known thrombophilia (antiphospholipid syndrome) were followed through pregnancy in an ethically approved prospective observational study. All women had dual energy X-ray absorptiometry (DEXA) scans at the lumbar spine (L1-L4) performed within 6 months prior to conception and in the immediate post-natal period, within 6 weeks of delivery. LMWH (5000 U/day) plus low-dose aspirin was commenced after a positive urine pregnancy test and continued throughout pregnancy and after delivery until 6 weeks post-partum. A group of 20 volunteers with recurrent miscarriage, not requiring any treatment intervention, acted as controls and were monitored in an identical fashion. RESULTS: Characteristics were not significantly different between treated patients and controls. Both groups showed a similar loss in lumbar spine (L1-L4) BMD by the end of the pregnancy [LMWH 4.17% or 0.045 g/cm(3), 95% confidence interval (CI) 0.036-0.062 versus control 3.56% or 0.043 g/cm(3), 95% CI 0.027-0.059]. However, the difference in bone loss between the groups was not statistically significant (0.002 g/cm(3), CI -0.0124 to 0.00865; P = 0.88). No patient suffered vertebral fracture. CONCLUSIONS: Bone loss associated with the use of long-term LMWH is not significantly different from physiological losses during pregnancy. PMID- 15121734 TI - Sperm autoimmunity. PMID- 15121735 TI - Reproductive history and Creutzfeld-Jakob disease. PMID- 15121736 TI - Creutzfeldt-Jakob disease and urinary gonadotrophins. PMID- 15121737 TI - Interindividual variation in abdominal subcutaneous and visceral adipose tissue: influence of measurement site. AB - We evaluated the influence of measurement site on the ranking (low to high) of abdominal subcutaneous (SAT) and visceral (VAT) adipose tissue. We also determined the influence of measurement site on the prediction of abdominal SAT and VAT mass. The subjects included 100 men with computed tomography (CT) measurements at L4-L5 and L3-L4 levels and 100 men with magnetic resonance imaging (MRI) measurements at L4-L5 and 5 cm above L4-L5 (L4-L5 +5 cm). Corresponding mass values were determined by using multiple-image protocols. For SAT, 90 and 92 of the 100 subjects for CT and MRI, respectively, had a difference in rank position at the two levels. The change in rank position exceeded the error or measurement for approximately 75% of the subjects for both methods. For VAT, 91 and 95 of the 100 subjects for CT and MRI, respectively, had a difference in rank position at the two levels. The change in rank position exceeded the error of measurement for 36% of the subjects for CT and for 8% of the subjects for MRI. For both imaging modalities, the variance explained in SAT and VAT mass (kg) was comparable for L4-L5, L4-L5 +5 cm, and L3-L4 levels. In conclusion, the ranking of subjects for abdominal SAT and VAT quantity is influenced by measurement location. However, the ability to predict SAT and VAT mass by using single images obtained at the L4-L5, L4-L5 +5 cm, or L3-L4 levels is comparable. PMID- 15121738 TI - Probing the impact of axial diffusion on nitric oxide exchange dynamics with heliox. AB - Exhaled nitric oxide (NO) is a potential noninvasive index of lung inflammation and is thought to arise from the alveolar and airway regions of the lungs. A two compartment model has been used to describe NO exchange; however, the model neglects axial diffusion of NO in the gas phase, and recent theoretical studies suggest that this may introduce significant error. We used heliox (80% helium, 20% oxygen) as the insufflating gas to probe the impact of axial diffusion (molecular diffusivity of NO is increased 2.3-fold relative to air) in healthy adults (21-38 yr old, n = 9). Heliox decreased the plateau concentration of exhaled NO by 45% (exhalation flow rate of 50 ml/s). In addition, the total mass of NO exhaled in phase I and II after a 20-s breath hold was reduced by 36%. A single-path trumpet model that considers axial diffusion predicts a 50% increase in the maximum airway flux of NO and a near-zero alveolar concentration (Ca(NO)) and source. Furthermore, when NO elimination is plotted vs. constant exhalation flow rate (range 50-500 ml/s), the slope has been previously interpreted as a nonzero Ca(NO) (range 1-5 ppb); however, the trumpet model predicts a positive slope of 0.4-2.1 ppb despite a zero Ca(NO) because of a diminishing impact of axial diffusion as flow rate increases. We conclude that axial diffusion leads to a significant backdiffusion of NO from the airways to the alveolar region that significantly impacts the partitioning of airway and alveolar contributions to exhaled NO. PMID- 15121739 TI - Hypoxia-responsive growth factors upregulate periostin and osteopontin expression via distinct signaling pathways in rat pulmonary arterial smooth muscle cells. AB - This study tested the hypothesis that expression of the novel adhesion molecule periostin (PN) and osteopontin (OPN) is increased in lung and in isolated pulmonary arterial smooth muscle cells (PASMCs) in response to the stress of hypoxia and explored the signaling pathways involved. Adult male rats were exposed to 10% O2 for 2 wk, and growth-arrested rat PASMCs were incubated under 1% O2 for 24 h. Hypoxia increased PN and OPN mRNA expression in rat lung. In PASMCs, hypoxia increased PN but not OPN expression. The hypoxia-responsive growth factors fibroblast growth factor-1 (FGF-1) and angiotensin II (ANG II) caused dose- and time-dependent increases in PN and OPN expression in PASMCs. FGF 1-induced PN expression was blocked by the FGF-1 receptor antagonist PD-166866 and by inhibitors of phosphatidylinositol 3-kinase (PI3K) (LY-294002, wortmannin), p70S6K (rapamycin), MEK1/2 (U-0126, PD-98059), and p38MAPK (SB 203580) but not of JNK (SP-600125). ANG II-induced PN expression was blocked by the AT(1)-receptor antagonist losartan and by inhibitors of PI3K and MEK1/2. In contrast, FGF-1-induced OPN expression was blocked by inhibitors of JNK or MEK1/2 but not of PI3K, p70S6K, or p38MAPK. Activation of p70S6K and p38MAPK by anisomycin robustly stimulated PN but not OPN expression. This study is the first to demonstrate that growth factor-induced expression of PN in PASMCs is mediated through PI3K/p70S6K, Ras/MEK1/2, and Ras/p38MAPK signaling pathways, whereas the expression of OPN is mediated through Ras/MEK1/2 and Ras/JNK signaling pathways. These differences in signaling suggest that PN and OPN may play different roles in pulmonary vascular remodeling under pathophysiological conditions. PMID- 15121740 TI - Age- and fitness-related differences in limb venous compliance do not affect tolerance to maximal lower body negative pressure in men and women. AB - Aging and chronic exercise training influence leg venous compliance. Venous compliance affects responses to an orthostatic stress; its effect on tolerance to maximal lower body negative pressure (LBNP) in the elderly is unknown. The purpose of this study was to determine the influence of age and fitness, a surrogate measure of exercise training, on calf venous compliance and tolerance to maximal LBNP in men and women. Forty participants, 10 young fit (YF; age = 22.6 +/- 0.5 yr, peak oxygen uptake = 57.1 +/- 2.0 ml.kg(-1).min(-1)), 10 young unfit (YU; 23.1 +/- 1.0 yr, 41.1 +/- 2.0 ml.kg(-1).min(-1)), 10 older fit (OF; 73.9 +/- 2.0 yr, 39.0 +/- 2.0 ml.kg(-1).min(-1)), and 10 older unfit (OU; 70.9 +/ 1.6 yr, 27.1 +/- 2.0 ml.kg(-1).min(-1)), underwent graded LBNP to presyncope or 4 min at -100 mmHg. By utilizing venous occlusion plethysmography, calf venous compliance was determined by using the first derivative of the pressure-volume relation during cuff pressure reduction. We found that the more fit groups had greater venous compliance than their unfit peers (P < 0.05) as did the young groups compared with their older peers (P < 0.05) such that OU < YU = OF < YF. LBNP tolerance did not differ between groups. In conclusion, these data suggest that aging reduces, and chronic exercise increases, venous compliance. However, these data do not support a significant influence of venous compliance on LBNP tolerance. PMID- 15121741 TI - Effect of endurance training on muscle TCA cycle metabolism during exercise in humans. AB - We tested the theory that links the capacity to perform prolonged exercise with the size of the muscle tricarboxylic acid (TCA) cycle intermediate (TCAI) pool. We hypothesized that endurance training would attenuate the exercise-induced increase in TCAI concentration ([TCAI]); however, the lower [TCAI] would not compromise cycle endurance capacity. Eight men (22 +/- 1 yr) cycled at approximately 80% of initial peak oxygen uptake before and after 7 wk of training (1 h/day, 5 days/wk). Biopsies (vastus lateralis) were obtained during both trials at rest, after 5 min, and at the point of exhaustion during the pretraining trial (42 +/- 6 min). A biopsy was also obtained at the end of exercise during the posttraining trial (91 +/- 6 min). In addition to improved performance, training increased (P < 0.05) peak oxygen uptake and citrate synthase maximal activity. The sum of four measured TCAI was similar between trials at rest but lower after 5 min of exercise posttraining [2.7 +/- 0.2 vs. 4.3 +/- 0.2 mmol/kg dry wt (P < 0.05)]. There was a clear dissociation between [TCAI] and endurance capacity because the [TCAI] at the point of exhaustion during the pretraining trial was not different between trials (posttraining: 2.9 +/- 0.2 vs. pretraining: 3.5 +/- 0.2 mmol/kg dry wt), and yet cycle endurance time more than doubled in the posttraining trial. Training also attenuated the exercise-induced decrease in glutamate concentration (posttraining: 4.5 +/- 0.7 vs. pretraining: 7.7 +/- 0.6 mmol/kg dry wt) and increase in alanine concentration (posttraining: 3.3 +/- 0.2 vs. pretraining: 5.6 +/- 0.3 mmol/kg dry wt; P < 0.05), which is consistent with reduced carbon flux through alanine aminotransferase. We conclude that, after aerobic training, cycle endurance capacity is not limited by a decrease in muscle [TCAI]. PMID- 15121742 TI - Satellite cell regulation of muscle mass is altered at old age. AB - Muscle mass is decreased with advancing age, likely due to altered regulation of muscle fiber size. This study was designed to investigate cellular mechanisms contributing to this process. Analysis of male Fischer 344 X Brown Norway rats at 6, 20, and 32 mo of age demonstrated that, even though significant atrophy had occurred in soleus muscle by old age, myofiber nuclear number did not change, resulting in a decreased myonuclear domain. Also, the number of centrally located nuclei was significantly elevated in soleus muscle of 32-mo-old rats, correlating with an increase in gene expression of MyoD and myogenin. Whereas total 5'-bromo 2'deoxyuridine (BrdU)-positive nuclei were decreased at older ages, BrdU-positive myofiber nuclei were increased. These results suggest that, with age, loss of muscle mass is accompanied by increased myofiber nuclear density that involves fusion of proliferative satellite cells, resembling ongoing regeneration. Interestingly, centrally located myofiber nuclei were not BrdU labeled. Rats were subjected to hindlimb suspension (HS) for 7 or 14 days and intermittent reloading during HS for 1 h each day (IR) to investigate how aging affects the response of soleus muscle to disuse and an atrophy-reducing intervention. After 14 days of HS, soleus muscle size was decreased to a similar extent at all three ages. However, myofiber nuclear number and the total number of BrdU-positive nuclei decreased with HS only in the young rats. IR was associated with an attenuation of atrophy in soleus muscles of 6- and 20- but not 32-mo-old rats. Furthermore, IR was associated with an increase in BrdU-positive myofiber nuclei only in young rats. These data indicate that altered satellite cell function with age contributes to the impaired response of soleus muscle to an intervention that attenuates muscle atrophy in young animals during imposed disuse. PMID- 15121744 TI - M-wave properties during progressive motor unit activation by transcutaneous stimulation. AB - The aim of this study was to interpret changes in experimentally recorded M waves with progressive motor unit (MU) activation based on simulation of the surface electromyogram. Activation order during transcutaneous electrical stimulation was analyzed by investigating M-wave average rectified value, spectral properties, and conduction velocity (CV) during electrically elicited contractions. M-waves were detected from the biceps brachii muscle of 10 healthy male subjects by a linear adhesive array of eight electrodes. Electrical stimulation was delivered to the motor point at either constant current intensity (40, 60, 80, and 100% of the supramaximal stimulation current) or with linearly increasing current. A model of surface electromyogram generation that varied activation order based on MU size and location was used to interpret the experimental results. From the experimental and model analysis, it was found that 1) MUs tended to be activated from low to high CV and from the superficial to the deep muscle layers with increasing transcutaneous electrical stimulation of the biceps brachii muscle, and 2) characteristic spectral frequencies of the M-wave were affected by many factors other than average CV (such as the activation order by MU location or the spread of the MU innervation zones and CVs), thus decreasing with a concomitant increase in CV during progressive MU activation. PMID- 15121743 TI - MCT1 confirmed in rat striated muscle mitochondria. AB - We sought to test the hypothesis that monocarboxylate transporter isoform 1 (MCT1) is the inner mitochondrial membrane lactate/pyruvate transporter, and, as such, contributes to functioning of the intracellular lactate shuttle. However, presence of a mammalian mitochondrially localized MCT1 (mMCT1) has been contested. We sought to confirm by Western blotting the mitochondrial localization of MCT1 in rat cardiac, soleus, and extensor digitorum longus muscles utilizing three different cell fractionation methods and three different antibodies. We performed Western blotting using antibodies to cell membrane glucose transporter isoform GLUT1, inner mitochondrial constituent cytochrome oxidase, the monocarboxylate transporter protein chaperone CD147, as well as custom and commercially available MCT1 antibodies. Western blots demonstrated similar results with each MCT1 antibody and two of three methods of fractionation. MCT1 was found in the mitochondria, as well as in the sarcolemmal membrane and whole muscle homogenates. Probing with GLUT1 and CD147 demonstrated that mitochondrial fractions were not contaminated with sarcolemmal remnants. Probing with cytochrome oxidase showed mitochondrial localization of MCT1. Comparison of these results to the findings of others indicates that the most likely source of discrepancy is the cell fractionation procedure utilized. PMID- 15121745 TI - Daily short-period gravitation can prevent functional and structural changes in arteries of simulated microgravity rats. AB - This study was designed to clarify whether simulated microgravity-induced differential adaptational changes in cerebral and hindlimb arteries could be prevented by daily short-period restoration of the normal distribution of transmural pressure across arterial vasculature by either dorsoventral or footward gravitational loading. Tail suspension (Sus) for 28 days was used to simulate cardiovascular deconditioning due to microgravity. Daily standing (STD) for 1, 2, or 4 h, or +45 degrees head-up tilt (HUT) for 2 or 4 h was used to provide short-period dorsoventral or footward gravitational loading as countermeasure. Functional studies showed that Sus alone induced an enhancement and depression in vasoconstrictor responsiveness of basilar and femoral arterial rings, respectively, as previously reported. These differential functional alterations can be prevented by either of the two kinds of daily gravitational loading treatments. Surprisingly, daily STD for as short as 1 h was sufficient to prevent the differential functional changes that might occur due to Sus alone. In morphological studies, the effectiveness of daily 4-h HUT or 1-h STD in preventing the differential remodeling changes in the structure of basilar and anterior tibial arteries induced by Sus alone was examined by histomorphometry. The results showed that both the hypertrophic and atrophic changes that might occur, respectively, in cerebral and hindlimb arteries due to Sus alone were prevented not only by daily HUT for 4 h but also by daily STD even for 1 h. These data indicate that daily gravitational loading by STD for as short as 1 h is sufficient to prevent differential adaptational changes in function and structure of vessels in different anatomic regions induced by a medium-term simulated microgravity. PMID- 15121746 TI - Influence of vehicle resistance on transdermal iontophoretic delivery of acetylcholine and sodium nitroprusside in humans. AB - Iontophoresis is a valuable method of noninvasive drug delivery for assessment of skin microvascular function, but it is important to consider and minimize its potential nonspecific electrical effects on blood flow. The use of sodium chloride (NaCl) instead of water as the iontophoresis vehicle has been reported to reduce these effects because it has a lower electrical resistance. However, this argument may not be valid when an agonist is added to the vehicle because its resistance will be changed. The aim of our study was to determine whether there is a difference in resistance between water and NaCl when used as vehicles for iontophoresis of acetylcholine (ACh) and sodium nitroprusside (SNP). Four cumulative doses of each drug, dissolved in either water or NaCl, were delivered via iontophoresis to the forearm skin of 14 healthy volunteers. We measured the resulting blood flow responses by using laser-Doppler imaging and the voltage across the electrodes for each delivery as an index of resistance. For ACh and SNP, there were no significant differences between the voltages measured when either water or NaCl was used as the vehicle. However, the blood flow responses to both agonists were significantly lower with NaCl (ACh: 25% lower, P < 0.001; SNP: 15% lower, P = 0.019). The use of NaCl is therefore unlikely to decrease any nonspecific electrical effects, and it may in fact reduce the effective dose of drug delivered. Deionized water is a better iontophoresis vehicle for the assessment of microvascular function in skin when using ACh and SNP. PMID- 15121747 TI - Does exercise-induced hypoxemia modify lactate influx into erythrocytes and hemorheological parameters in athletes? AB - This study investigated 1) red blood cells (RBC) rigidity and 2) lactate influxes into RBCs in endurance-trained athletes with and without exercise-induced hypoxemia (EIH). Nine EIH and six non-EIH subjects performed a submaximal steady state exercise on a cyclo-ergometer at 60% of maximal aerobic power for 10 min, followed by 15 min at 85% of maximal aerobic power. At rest and at the end of exercise, arterialized blood was sampled for analysis of arterialized pressure in oxygen, and venous blood was drawn for analysis of plasma lactate concentrations and hemorheological parameters. Lactate influxes into RBCs were measured at three labeled [U-14C]lactate concentrations (1.6, 8.1, and 41 mM) on venous blood sampled at rest. The EIH subjects had higher maximal oxygen uptake than non-EIH (P < 0.05). Total lactate influx was significantly higher in RBCs from EIH compared with non-EIH subjects at 8.1 mM (1,498.1 +/- 87.8 vs. 1,035.9 +/- 114.8 nmol.ml(-1).min(-1); P < 0.05) and 41 mM (2,562.0 +/- 145.0 vs. 1,618.1 +/- 149.4 nmol.ml(-1).min(-1); P < 0.01). Monocarboxylate transporter-1-mediated lactate influx was also higher in EIH at 8.1 mM (P < 0.05) and 41 mM (P < 0.01). The drop in arterial oxygen partial pressure was negatively correlated with total lactate influx measured at 8.1 mM (r = -0.82, P < 0.05) and 41 mM (r = -0.84, P < 0.05) in the two groups together. Plasma lactate concentrations and hemorheological data were similar in the two groups at rest and at the end of exercise. The results showed higher monocarboxylate transporter-1-mediated lactate influx in the EIH subjects and suggested that EIH could modify lactate influx into erythrocyte. However, higher lactate influx in EIH subjects was not accompanied by an increase in RBC rigidity. PMID- 15121748 TI - Adaptation to lengthening contraction-induced injury in mouse muscle. AB - Adaptations to repeated bouts of injury-inducing lengthening contractions were studied in mouse anterior crural muscles. Five bouts of 150 lengthening contractions were performed in vivo, with each bout separated by 2 wk of rest. Three primary observations were made. First, there was little, if any, attenuation in the immediate isometric torque losses after lengthening contractions at "physiological" stimulation frequencies (i.e., <125 Hz), although there was a pronounced decrease in torque loss at higher frequencies between the first and second bouts. Second, the immediate losses in strength that occurred after all five lengthening contraction bouts could be explained in part by excitation-contraction uncoupling. Third, the most important adaptation was a significant enhancement in the rate of recovery of strength after the lengthening contractions. It is probable that the accelerated rate of strength recovery resulted from the more rapid loss and subsequent recovery of myofibrillar protein observed after the fifth bout. PMID- 15121749 TI - Repetitive measurements of pulmonary mechanics to inhaled cholinergic challenge in spontaneously breathing mice. AB - Precise and repeatable measurements of pulmonary function in intact mice are becoming increasingly important for experimental investigations on various respiratory disorders including asthma. Here, we present validation of a novel in vivo method that, for the first time, combines direct and repetitive recordings of standard pulmonary mechanics with cholinergic aerosol challenges in anesthetized, orotracheally intubated, spontaneously breathing mice. We demonstrate that, in several groups of nonsensitized BALB/c mice, dose-related increases in pulmonary resistance and dynamic compliance to aerosolized methacholine are reproducible over short and extended intervals without causing detectable cytological alterations in the bronchoalveolar lavage or relevant histological changes in the proximal trachea and larynx regardless of the number of orotracheal intubations. Moreover, as further validation, we confirm that allergic mice, sensitized and challenged with Aspergillus fumigatus, were significantly more responsive to cholinergic challenge (P < 0.01) and exhibited marked eosinophilia and lymphocytosis in bronchoalveolar lavage fluids as well as significant pathological alterations in laryngotracheal histology compared with nonsensitized mice. We suggest that this approach will provide useful and necessary information on pulmonary mechanics in studies of various respiratory disorders in mice, including experimental models of asthma and chronic obstructive pulmonary disorder, investigations of pulmonary pharmacology, or more general investigations of the genetic determinants of lung function. PMID- 15121751 TI - Magnetic Resonance for T-staging of nasopharyngeal carcinoma--the most informative pair of sequences. AB - OBJECTIVE: To evaluate the most informative pair of sequences in magnetic resonance (MR) for T-staging of nasopharyngeal carcinoma (NPC). METHODS: The MR images of 134 patients with newly diagnosed NPC, from 1996 to 2002, were retrospectively reviewed. All the patients were scanned using 1.5 Tesla MR systems. The images of the nasopharynx were reviewed by two qualified radiologists to determine the positive findings and the T-stage by UICC (6th edition) System, using each sequence separately. The T-stage derived from a single MR sequence was then compared with the T-stage based on the five selected sequences to assess the number and percentage of patients who were being understaged. Therefore, the overall percentage accuracy of each single sequence could be determined. A pair of sequences providing information to achieve almost 100% diagnostic accuracy was then derived. RESULTS: The overall percentage accuracy of five individual sequences of the nasopharynx is as follows: contrast enhanced (CE) fat suppression (FS) axial T1 (94.8%), CE FS coronal T1 (88.1%), FS axial T2 (85.8%), non-contrast enhanced (NE) axial T1 (78.4%) and non-contrast enhanced (NE) coronal T1 (77.6%). CE FS axial T1 has the best accuracy. All the structures that are missed in CE FS axial T1, which lead to apparent understaging, are appreciated in NE axial T1-weighted images. CONCLUSION: Individual sequences supplement each other in the NPC staging. CE FS axial T1 is the most informative individual sequence. Combination of CE FS axial T1 and NE axial T1 of the nasopharynx provides sufficient information to achieve almost 100% diagnostic accuracy in T-staging; therefore, both should be included in the MR-staging protocol. PMID- 15121752 TI - Involvement of viral and chemical factors with oral cancer in Taiwan. AB - BACKGROUND: The association between oral squamous cell carcinoma (OSCC) and viral and chemical factors is uncertain. Therefore the correlation of viral and chemical factors with oral cancer in Taiwan was investigated. METHODS: Thirty seven paraffin-embedded oral cancer biopsies and 36 normal oral tissue specimens were examined by the polymerase chain reaction method for six viruses: HPV, CMV, EBV, HSV-1, HSV-2 and HHV-8. To elucidate the role of arecoline in the oncogenesis of oral cancer, human buccal fibroblasts, oral submucosal fibroblasts and three cancer cell lines KB, GNM and TSCCa were used for MTT cytotoxity assay and flow cytometry DNA content analysis. RESULTS: Two (5.4%) HSV-1-positive and four (10.8%) HPV-positive cases were recognized in oral cancer biopsies. Among the four HPV-positive tissues, two were further typed as HPV-16, one was identified as HPV-18- and HSV-1-positive; and one contained both HPV-16 and HPV 18. One sample presented HSV-1 only. Arecoline, at a concentration lower than 0.8 micro g/ml, increased cell growth (all cell types); at higher concentrations (25 400 micro g/ml) it was cytotoxic. The cell cycle was demonstrated to be altered either by low or high concentrations of arecoline treatment, depending on the cells treated. CONCLUSIONS: The data demonstrated that HPV, HSV-1 and betel quid chewing were significantly associated with OSCC, but HSV-2, CMV, EBV and HHV-8 were not. We suggest that the most determinative factor for oral cancer may be chemical in nature rather than viral infection. PMID- 15121753 TI - Serum vascular endothelial growth factor per platelet count in hepatocellular carcinoma: correlations with clinical parameters and survival. AB - BACKGROUND: Platelets have been reported to act as transporters of tumor originated vascular endothelial growth factor (VEGF), contributing to tumor angiogenesis and progression. Serum VEGF per platelet count, as an indirect theoretical estimate of VEGF in platelets, may predict the malignant potential of tumors. However, its prognostic significance is still unclear in hepatocellular carcinoma (HCC), a highly vascular tumor. METHODS: Serum VEGF was measured by enzyme-linked immunosorbent assay. We compared serum VEGF, platelet count and serum VEGF per platelet count in 52 HCC patients, 26 liver cirrhosis patients and 30 healthy controls. The relation of serum VEGF per platelet count with clinicopathologic variables of HCC patients and the prognostic significance were investigated. RESULTS: Serum VEGF per platelet count in HCC patients was higher than in liver cirrhosis patients and healthy controls (P < 0.01). There was a statistically significant correlation between serum VEGF and platelet count in HCC patients (r = 0.751, P < 0.01). Serum VEGF per platelet count was higher in patients with advanced stage and portal vein thrombosis (P < 0.01). Patients with high serum VEGF per platelet count (>1.4 pg/10(6)) showed poor response to treatment and shorter overall survival (P < 0.01). Serum VEGF per platelet count was an independent prognostic factor with the presence of portal vein thrombosis (P < 0.01). CONCLUSIONS: Serum VEGF per platelet count could be a feasible prognostic indicator during the follow-up of patients with HCC. PMID- 15121754 TI - Hypermethylation-associated inactivation of the SOCS-1 gene, a JAK/STAT inhibitor, in human pancreatic cancers. AB - BACKGROUND: SOCS-1, a JAK-binding protein (SSI-1/SOCS-1/JAB), regulates the JAK/STAT signal transduction pathway that relays signals from various cytokines in the extracellular matrix into the cell. Inactivation of the SOCS-1 gene by methylation has been previously described in hepatocellular carcinomas and multiple myeloma. The purpose of the present work was to analyze the expression of the SOCS-1 gene and identify inactivation of this gene by methylation in pancreatic cancers. METHODS: 20 samples were analyzed. We identified the expression of SOCS-1 gene using RT-PCR and the mechanism of inactivation in this gene by methylation assay. RESULTS: We documented marked suppression of SOCS-1 mRNA and reduction of SOCS-1 protein in 7 of 14 primary pancreatic cancers examined; moreover, CpG-rich regions upstream of the SOCS-1 gene were hypermethylated in 8 of the 14 tumors. CONCLUSIONS: The results suggested that this gene is silenced in a substantial portion of pancreatic cancers through mechanisms that cause methylation in the promoter region. PMID- 15121755 TI - A pilot phase II study of capecitabine in advanced or recurrent colorectal cancer. AB - BACKGROUND: A pilot phase II study was conducted to evaluate the Japanese intermittent regimen of capecitabine in patients with advanced/recurrent colorectal cancer. METHODS: Twenty-two patients received oral capecitabine in a dose of 828 mg/m(2) twice daily for 3 weeks every 4 weeks. RESULTS: In the 20 patients evaluable for efficacy, the overall response rate was 25.0% (95% CI, 8.7 49.1%), rising to 33.0% in the subset of patients previously untreated for metastatic disease (n = 9). A further nine patients had stable disease. The median duration of response was 7.0 months. Five patients (22.7%) experienced grade 3/4 treatment-related adverse events, the most common being a bullous rash observed in two patients (9.1%). CONCLUSIONS: The 3 weeks out of 4 intermittent regimen of capecitabine demonstrated good antitumor activity and tolerability in patients with advanced/refractory colorectal cancer, providing a clear rationale for conducting a larger phase II study in patients with advanced disease. PMID- 15121756 TI - 15-year experience on intravesical therapy of T1G3 urinary bladder cancer: a conservative approach. AB - OBJECTIVE: To report the recurrence, progression and survival in patients with T1G3 transitional cell carcinoma (TCC) of the urinary bladder treated with sequential intravesical bacillus Calmette-Guerin (BCG) and chemotherapeutic agents (doxorubicin or epirubicin) on long-term follow up. METHODS: Between July 1988 and September 1999, all patients in a single center with T1G3 bladder TCC, after complete transurethral resection (TURBT), received either 81 mg of Connaught strain BCG or 50 mg of doxorubicin or epirubicin as adjuvant therapy. A conservative approach was adopted whereby those with superficial recurrences were eligible to crossover, even repeatedly, until progression to muscle invasion. Recurrence, progression and disease-specific survival were analyzed. RESULTS: There were 36 patients included, with 26 males and 10 females. The mean age was 71.6 years (range 53-85 years). Final analysis was made at a median follow-up of 23.5 months (range 0-125 months) for recurrence, 33 months (range 0-125 months) for progression and 45.5 months (range 3-125 months) for survival. Sixteen (44.4%) patients showed recurrence. Nine (25%) of these 16 patients progressed. Five (13.9%) of those who progressed died of TCC. The 10 year Kaplan-Meier estimates for recurrence-free survival, progression-free survival and disease specific survival were 48, 68 and 81%, respectively. Figures with this conservative approach were comparable to those with more aggressive approaches reported in the literature. CONCLUSIONS: Adjuvant intravesical therapy with either BCG or a chemotherapeutic agent (doxorubicin or epirubicin) and crossover on recurrence was an effective conservative treatment for T1G3 bladder TCC. PMID- 15121757 TI - High body mass index correlates with increased risk of venous irritation by vinorelbine infusion. AB - BACKGROUND: Vinorelbine is currently one of the most active chemotherapeutic agents. However, it is also a moderate vesicant that is well known to cause venous irritation and phlebitis. We conducted this study to identify clinical risk factors related to the incidence of venous irritation caused by peripheral vinorelbine infusion. METHODS: Medical records were used to investigate retrospectively a total of 201 cases of non-small cell lung cancer treated with a chemotherapeutic regimen containing vinorelbine. Venous irritation was evaluated in every course and graded according to the National Cancer Institute Common Toxicity Criteria version 2.0. Gender, age, body mass index (BMI), chemotherapeutic regimen, dose of vinorelbine and prior chemotherapy were used as clinical variables. RESULTS: A total of 928 vinorelbine infusions were administered to the 201 patients, among whom venous irritation occurred in 63 (31%). The incidence of venous irritation was 28% in the normal BMI (<25) group and 45% in the high BMI (25 or more) group and the difference between the two groups was statistically significant (P = 0.037). There were no significant correlations between the incidence of venous irritation and the clinical variables except BMI. In the multivariate analysis BMI was also a significant independent variable that correlated with increased risk of venous irritation (P = 0.017). CONCLUSIONS: Care is required when using vinorelbine to treat patients with a high BMI, especially with regard to the development of venous irritation. PMID- 15121758 TI - Skeletal muscle metastases of carcinoma: a clinicopathological study of 12 cases. AB - OBJECTIVE: To clarify the clinical and magnetic resonance (MR) imaging features of a rare condition of metastasis of carcinoma to skeletal muscle. METHOD: Clinicopathological findings for 12 patients (10 male, two female, age range 48 89 years, mean age 68 years) with skeletal muscle metastases of carcinomas were reviewed retrospectively. RESULTS: In nine of the 12 patients the skeletal muscle metastasis was presented as "painful mass". The lung was found to be the most common primary source, accounting for 33% of the cases, and the lower extremity was the most common metastatic site, accounting for 67% of the current series. Diagnosis was made by biopsy in all cases. Overall, MR images were not specific, but on the gadolinium-DTPA enhanced MR images, extensive peritumoral enhancement associated with central necrosis was found in 11 of the 12 patients (92%). Seven patients died within 2-19 months (average: 9 months) after the detection of the skeletal muscle metastasis, among whom only one patient was continuously disease free for 92 months after wide excision of the metastatic lesion. CONCLUSION: Skeletal muscle metastasis is often presented as a painful mass in patients with known primary carcinoma. For diagnosis, needle biopsy is mandatory. However, a painful mass with an extensive peritumoral enhancement should be highly suspected to represent carcinoma metastasis to skeletal muscles. In selected patients, wide excision with combined chemotherapy could yield unexpectedly good results. PMID- 15121759 TI - Successful pregnancy and delivery in a patient with chronic myelogenous leukemia (CML), and management of CML with leukapheresis during pregnancy: a case report and review of the literature. AB - Although infrequently seen, the management of cancer during pregnancy can be difficult for patients, their families and physicians. The concomitant occurrence of pregnancy and chronic myelogenous leukemia is uncommon. We describe the successful management of a 26-year-old woman in the first trimester of her pregnancy with chronic myelogenous leukemia (CML) in chronic phase by using only leukapheresis. She was treated with leukapheresis until her delivery at 36 weeks of gestation. The procedure was without significant adverse effects on the patient or fetus. We applied a total of 15 leukapheresis treatments throughout the pregnancy. The patient gave birth vaginally to a healthy 2800 g boy at 36 weeks of gestation. We conclude that leukapheresis may provide an alternative treatment to chemotherapy, alpha-interferon or imatinib in pregnant patients with CML, particularly with concern over their potential teratogenic and other adverse effects. PMID- 15121760 TI - The orphan nuclear receptor LRH-1 activates the ABCG5/ABCG8 intergenic promoter. AB - The ATP binding cassette (ABC) half-transporters ABCG5 and ABCG8 facilitate biliary and intestinal removal of neutral sterols. Here, we identify a binding site for the orphan nuclear receptor liver receptor homolog-1 (LRH-1) at nt 134 142 of the ABCG5/ABCG8 intergenic region necessary for the activity of both the ABCG5 and ABCG8 promoters. Mutating this LRH-1 binding site reduced promoter activity of the human ABCG5/ABCG8 intergenic region more than 7-fold in HepG2 and Caco2 cells. Electrophoretic mobility shift assays with HepG2 nuclear extracts demonstrated specific binding of LRH-1 to the LRH-1 binding motif in the human ABCG5/ABCG8 intergenic region. LRH-1 overexpression increased promoter activity up to 1.6-fold and 3-fold in Caco2 and 293 cells, respectively. Finally, deoxycholic acid repressed the ABCG5 and ABCG8 promoters, consistent with bile acid regulation via the farnesoid X receptor-small heterodimeric partner-LRH-1 pathway. These results demonstrate that LRH-1 is a positive transcription factor for ABCG5 and ABCG8 and, in conjunction with studies on LRH-1 activation of other promoters, identify LRH-1 as a "master regulator" for genes involved in sterol and bile acid secretion from liver and intestine. PMID- 15121761 TI - Mechanism of action of galantamine on N-methyl-D-aspartate receptors in rat cortical neurons. AB - Galantamine, a new Alzheimer's drug approved in the United States, is known to inhibit acetylcholinesterase and potentiate acetylcholine-induced currents in brain neurons. However, because both cholinergic and N-methyl-D-aspartate (NMDA) systems are down-regulated in the brain of Alzheimer's patients, we studied the effects of galantamine on NMDA receptors. NMDA-induced whole-cell currents were recorded from the rat multipolar cortical neurons in primary culture. NMDA currents recorded in Mg2+-free media without addition of glycine were reversibly potentiated by bath and U-tube applications of galantamine at 10 to 10,000 nM, showing a bell-shaped dose-response relationship. However, alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionic acid and kainate currents were not affected by galantamine. The maximum potentiation of NMDA currents to approximately 130% of the control was obtained at 1 microM galantamine. The potentiation was due to a shift of the NMDA dose-response curve in the direction of lower NMDA concentrations. Glycine at 1 to 3000 nM enhanced NMDA currents, and potentiation by 1 microM galantamine and 1 to 300 nM glycine was additive. The glycine site antagonist 7-chlorokynurenic acid did not prevent the galantamine action. These results suggested that galantamine did not interact with the glycine binding site. Experiments with various concentrations of Mg2+ indicated that galantamine did not affect the Mg2+ blocking site of the NMDA receptor. PKC was involved in galantamine potentiation of NMDA currents, but protein kinase A, Gi/Go proteins, and Gs proteins were not involved. Potentiation of the activity of NMDA receptors is deemed partially responsible for the improvement of cognition, learning, and memory in Alzheimer's patients. PMID- 15121762 TI - Lobeline analogs with enhanced affinity and selectivity for plasmalemma and vesicular monoamine transporters. AB - Lobeline attenuates the behavioral effects of psychostimulants in rodents and inhibits the function of nicotinic receptors (nAChRs), dopamine transporters (DATs), and vesicular monoamine transporters (VMAT2s). Monoamine transporters are considered valid targets for drug development for the treatment of methamphetamine abuse. In the current study, a series of lobeline analogs were evaluated for affinity and selectivity at these targets. None of the analogs was more potent than nicotine at the [3H]methyllycaconitine binding site (alpha7* nAChR subtype). Lobeline tosylate was equipotent with lobeline in inhibiting [3H]nicotine binding but 70-fold more potent in inhibiting nicotine-evoked 86Rb+ efflux, demonstrating antagonism of alpha4beta2* nAChRs. Compared with lobeline, the defunctionalized analogs lobelane, mesotransdiene, and (-)-trans-transdiene showed dramatically reduced affinity at alpha4beta2* nAChRs and a 15- to 100-fold higher affinity (Ki = 1.95, 0.58, and 0.26 microM, respectively) at DATs. Mesotransdiene and (-)-trans-transdiene competitively inhibited DAT function, whereas lobelane and lobeline acted noncompetitively. 10S/10R-MEPP [N-methyl-2R (2R/2S-hydroxy-2-phenylethyl)6S-(2-phenylethyl)piperidine] and 10R-MESP [N-methyl 2R-(2R-hydroxy-2-phenylethyl)6S-(2-phenylethen-1-yl)piperidine] were 2 to 3 orders of magnitude more potent (Ki = 0.01 and 0.04 microM, respectively) than lobeline in inhibiting [3H]serotonin uptake; 10S/10R-MEPP showed a 600-fold selectivity for this transporter. Uptake results using hDATs and human serotonin transporters expressed in human embryonic kidney-293 cells were consistent with native transporter assays. Lobelane and ketoalkene were 5-fold more potent (Ki = 0.92 and 1.35 microM, respectively) than lobeline (Ki = 5.46 microM) in inhibiting [3H]methoxytetrabenazine binding to VMAT2 in vesicle preparations. Thus, structural modification (defunctionalization) of the lobeline molecule markedly decreases affinity for alpha4beta2* and alpha7* nAChRs while increasing affinity for neurotransmitter transporters, affording analogs with enhanced selectivity for these transporters and providing new leads for the treatment of psychostimulant abuse. PMID- 15121763 TI - Chronic antidepressant treatment causes a selective reduction of mu-opioid receptor binding and functional coupling to G Proteins in the amygdala of fawn hooded rats. AB - We have previously documented that chronic alcohol consumption or alcohol withdrawal affects mu-opioid receptor density and receptor-mediated G protein coupling in Fawn-Hooded (FH) rat brain, especially in mesolimbic regions. FH rats demonstrate comorbid depression and high voluntary alcohol consumption; treatment with standard antidepressants improves both facets of this phenotype. Accordingly, we sought to examine whether mu-opioid receptor binding and the receptor-mediated functional coupling to G protein is affected by this drug treatment. Using quantitative autoradiography, binding of mu-opioid receptors labeled by [125I]FK33,824 (D-Ala2,N-Me-Phe4,Met(O)5-ol enkephalin) and the coupling between receptors and G proteins determined by agonist-stimulated guanosine 5'-O -(3-[35S]thio)triphosphate ([35S]GTPgammaS) binding was mapped throughout brain sections of FH rats after 10-day treatment with vehicle, desipramine, or sertraline. Both desipramine and sertraline produced significant decreases of [125I]FK33,824 binding in many brain regions; 13 of 20 measured regions for desipramine and 16 of 20 measured regions for sertraline. The coupling efficiency of mu-opioid receptors to G proteins was determined by an increase of [35S]GTPgammaS binding induced by stimulation with the mu-opioid receptor agonist [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (10 microM). In contrast to the receptor binding profile, functional coupling of receptors to G proteins was only significantly reduced in the amygdala, whereas it remained unchanged in other regions compared with control. The present findings suggest that antidepressants regulate opioid systems; however, this occurs differentially, and region-specific alteration of functional coupling of mu-opioid receptors to G proteins in the amygdala suggests that opioid function within the amygdala may be modulated by antidepressants. PMID- 15121764 TI - Metabolism of N,N',N"-triethylenethiophosphoramide by CYP2B1 and CYP2B6 results in the inactivation of both isoforms by two distinct mechanisms. AB - The anticancer drug N,N,"N"-triethylenethiophosphoramide (tTEPA) inactivated CYP2B6 and CYP2B1 in the reconstituted system in a time-, concentration-, and NADPH-dependent manner indicative of mechanism-based inactivation. The KI value for the inactivation of CYP2B1 was 38 microM, the kinact was 0.3 min(-1), and the t1/2 value was 2.5 min. Spectral carbon monoxide (CO) binding and high performance liquid chromatography heme studies of the tTEPA-inactivated CYP2B1 suggest that the loss in the enzymatic activity was primarily due to the binding of a reactive tTEPA intermediate to the 2B1 apoprotein. Inactivation by tTEPA in the presence of 7-ethoxycoumarin, an alternate substrate, reduced the rate of inactivation of CYP2B1. Incubations with tTEPA and NADPH resulted in greater than 90% loss in the 7-ethoxy-4-(trifluoromethyl)coumarin O-deethylation and testosterone hydroxylation activity of CYP2B1. In contrast, benzphetamine metabolism was significantly less inhibited (47%). CYP2B6 was inactivated by tTEPA with a KI value of 50 microM, a k inact value of 0.1 min(-1), and a t1/2 value of 14 min. However, unlike CYP2B1, the tTEPA-inactivated human isoform showed losses in the cytochrome P450 (P450) CO spectrum, the pyridine hemochrome spectrum, and in the amount of native heme that were comparable with the loss in the 7-EFC and benzphetamine activity, suggesting that activity loss was brought about by a tTEPA-reactive intermediate damaging the CYP2B6 heme. CYP2B6 could only be protected from the tTEPA-dependent inactivation by the 2B6-specific substrate bupropion but not by other substrates of CYP2B such as benzphetamine, testosterone, or 7-ethoxycoumarin. The data indicate that tTEPA metabolism by these two 2B isoforms results in inactivation of the P450s by two distinct mechanisms. PMID- 15121765 TI - Effect of BM-573 [N-terbutyl-N'-[2-(4'-methylphenylamino)-5-nitro benzenesulfonyl]urea], a dual thromboxane synthase inhibitor and thromboxane receptor antagonist, in a porcine model of acute pulmonary embolism. AB - The aim of this study was to evaluate the effect of BM-573 [N-terbutyl-N'-[2-(4' methylphenylamino)-5-nitro-benzenesulfonyl]urea], a dual thromboxane A2 synthase inhibitor and receptor antagonist, on the hemodynamic response to acute pulmonary embolism. Six anesthetized pigs were infused with placebo (placebo group) and compared with six other pigs receiving a continuous infusion of BM-573 (BM group). Pulmonary embolization with 0.3 g/kg autologous blood clots was carried out 30 min after the start of the infusion. Right ventricular pressure-volume loops were recorded using a conductance catheter, and end-systolic ventricular elastance was periodically assessed by varying right ventricular preload. Pulmonary vascular properties were studied by use of a four-element windkessel model. Hemodynamic data, including assessment of right ventricular-arterial coupling, were collected at baseline and every 30 min for 4 h. Blood samples were collected to assess gas exchange, thromboxane A2, and prostacyclin plasma levels and to evaluate platelet aggregation. Mean pulmonary arterial pressure in the placebo group increased significantly more than in the BM group, mainly because of an additional increase in pulmonary vascular resistance. Arterial and end systolic ventricular elastances increased also more in the placebo group, whereas right ventricular efficiency decreased. BM-573 prevented both platelet aggregation induced by U-46619 (9,11-dideoxy-11alpha,9alpha epoxymethanoprostaglandin F2alpha) or by arachidonic acid, and thromboxane A2 overproduction, whereas prostacyclin liberation was preserved. Oxygenation, however, was not significantly improved. We conclude that in this animal model of acute pulmonary embolism, infusion of BM-573 reduced pulmonary vasoconstriction. As a result, right ventricular-vascular coupling values were maintained at a maximal efficiency level. PMID- 15121766 TI - Cardiovascular genetics: are we there yet? PMID- 15121767 TI - Will the real Cowden syndrome please stand up (again)? Expanding mutational and clinical spectra of the PTEN hamartoma tumour syndrome. PMID- 15121769 TI - Large scale association analysis for identification of genes underlying premature coronary heart disease: cumulative perspective from analysis of 111 candidate genes. AB - BACKGROUND: to date, only three groups have reported data from large scale genetic association studies of coronary heart disease using a case control design. METHODS AND RESULTS: to extend our initial report of 62 genes, we present data for 210 polymorphisms in 111 candidate genes genotyped in 352 white subjects with familial, premature coronary heart disease (onset age for men, 45; for women, 50) and a random sample of 418 population based whites. Multivariate logistic regression analysis was used to compare the distributions of genotypes between cases and the comparison group while controlling for age, sex, body mass, diabetes, and hypertension. Significant associations were found with polymorphisms in thrombospondin-4 (THBS4), thrombospondin-2 (THBS2) and plasminogen activator inhibitor-2 (PAI2), the strongest being with the A387P variant in THBS4 (p = 0.002). The THBS2 and THBS4 associations have since been replicated. We evaluated polymorphisms in 40 genes previously associated with coronary heart disease and found significant (p<0.05) associations with 10: ACE, APOE, F7, FGB, GP1BA, IL1RN, LRP1, MTHFR, SELP, and THPO. For five of these genes, the polymorphism associated in our study was different from that previously reported, suggesting linkage disequilibrium as an explanation for failure to replicate associations consistently across studies. We found strong linkage disequilibrium between polymorphisms within and between genes, especially on chromosome 1q22-q25, a region containing several candidate genes. CONCLUSIONS: despite known caveats of genetic association studies, they can be an effective means of hypothesis generation and complement classic linkage studies for understanding the genetic basis of coronary heart disease. PMID- 15121771 TI - Variable penetrance of a familial progressive necrotising encephalopathy due to a novel tRNA(Ile) homoplasmic mutation in the mitochondrial genome. AB - INTRODUCTION: We present a family comprising a clinically normal mother and two daughters, each with severe encephalopathy with onset in late childhood. A third daughter had died previously of an earlier onset but neuropathologically similar disease. METHODS: Sequence analysis of the entire mtDNA was carried out in muscle, fibroblasts, and lymphocytes of the affected daughters and unaffected mother. Biochemical analysis of individual respiratory chain enzymes was performed on the same tissues, and on several transmitochondrial cybrid clones containing the nucleus of a 143B.206 osteosarcoma cell line and the mutant mtDNA. RESULTS: Genetic analyses revealed in both daughters and mother the presence of a novel mutation in the tRNA(Ile) gene of mtDNA, which was homoplasmic in fibroblasts, lymphocytes, and skeletal muscle of the two patients. It was also homoplasmic in fibroblast and skeletal muscle samples of the mother, and approximately 97% heteroplasmic in her lymphocytes. Combined defects of complexes I and IV of the mitochondrial respiratory chain were found not only in fibroblasts of the two probands, but surprisingly also in those of their clinically unaffected mother. The respiratory chain defect segregated in transmitochondrial cybrids containing the nucleus of a 143B.206 osteosarcoma cell line and the mutant mtDNA, indicating that the latter was responsible for the biochemical phenotype. DISCUSSION: Our results support the concept that homoplasmic mutations in tRNA genes can be responsible for mitochondrial disorders characterised by extremely variable penetrance. Albeit still unexplained, this phenomenon has important consequences in the nosological characterisation, clinical management, and genetic counselling of mitochondrial disorders. PMID- 15121768 TI - Genotype-phenotype correlations in Peutz-Jeghers syndrome. AB - BACKGROUND AND AIMS: Peutz-Jeghers syndrome (PJS) is a dominantly inherited disorder often caused by mutations in STK11. Time to onset of symptoms was characterised for a large collection of individuals with PJS who had been tested for STK11 mutations and genotype-phenotype correlations were evaluated. METHODS: We characterised mutations in 42 independent probands and also used a historical cohort design to study 51 individuals with Peutz-Jeghers syndrome who had completed self-administered questionnaires. RESULTS: Mutations were detected in 22/32 (69%) probands with PJS and 0/10 probands referred to rule out PJS. Real time PCR analysis to quantitate DNA failed to detect any large deletions in PJS participants without STK11 mutations. The median time to onset for gastrointestinal symptoms or polypectomy was 13 years of age but showed a wide variability. Gastric polyps were frequent in PJS participants, with a median age at onset of 16 years. Individuals with missense mutations had a significantly later time to onset of first polypectomy (p = 0.04) and of other symptoms compared with those participants either with truncating mutations or no detectable mutation. CONCLUSION: STK11 mutation analysis should be restricted to individuals who meet PJS criteria or their close relatives. Direct sequencing of STK11 yields a high rate of point mutations in individuals who meet phenotypic PJS criteria. Individuals with missense mutations of STK11 typically had a later time to onset for PJS symptoms. The common occurrence of gastric polyps may facilitate chemopreventive studies for this disorder. PMID- 15121772 TI - Functional dimorphism of two hAgRP promoter SNPs in linkage disequilibrium. AB - The agouti related protein (AgRP) exerts its anabolic effects on food intake by antagonising the alpha-melanocyte stimulating hormone (alpha-MSH) at its receptors, melanocortin receptors 3 and 4 (MC3R and MC4R). A single nucleotide polymorphism (SNP) in the promoter of the human AgRP (hAgRP), -38C>T, was associated with low body fatness. The -38T allele that was associated with low body fatness also resulted in lower promoter activity. Here we report a novel SNP, -3019G>A, again in the promoter of hAgRP, which is in complete linkage disequilibrium (LD) with the -38C>T SNP (linked alleles: -3019A/-38T and -3019G/ 38C). Functional analyses in a human adrenal and two mouse hypothalamus cell lines showed that the -3019A allele had significantly higher promoter activity. Hence, the two linked alleles (-3019A and -38T) had opposite effects on promoter function and yet they were both associated with low body fatness. The region encompassing the -38C>T SNP had approximately 1000-fold higher activity than the region encompassing the -3019G>A SNP, potentially determining the net functional effect between these two SNPs. PMID- 15121773 TI - Inhibin alpha-subunit (INHA) gene and locus changes in paediatric adrenocortical tumours from TP53 R337H mutation heterozygote carriers. AB - The R337H TP53 mutation is a low-penetrance molecular defect that predisposes to adrenocortical tumour (ACT) formation in Brazilian and possibly other populations. Additional genetic defects may be responsible for the variable expression of ACTs in these cases. The inhibin alpha-subunit gene (INHA) on 2q33 qter has been implicated in mouse adrenocortical tumourigenesis. We studied 46 pediatric patients with ACTs from Brazil for INHA genetic alterations; 39 of these patients were heterozygous carriers of the R337H TP53 mutation. We first mapped the INHA gene by radiation hybrid analysis and determined 10 linked microsatellite markers in an area flanked by D2S1371 and D2S206 on 2q33-qter. These markers were then used for loss of heterozygozity (LOH) studies in nine paired germline and tumour DNA samples. Mapping placed the INHA gene in close proximity to D2S2848 (SHGC11864) with a log of odds (LOD) score of 5.84. LOH for at least one marker in the region was identified in 8/9 tumours (89%). Six patients were heterozygous for three INHA mutations: one in exon 1, 127C>G, and two in exon 2, 3998G>A and 4088G>A, all leading to amino acid substitutions (P43A, G227R, and A257T, respectively). A257T is located in a conserved INHA region, highly homologous to transforming growth factor-beta; both G227R and A257T change polarity, and, in addition, G227R changes the pH. We conclude that these sequence alterations and the detected 2q allelic changes suggest that INHA may be one of the contributing factors needed for ACT formation in pediatric patient carriers of the R337H TP53 mutation. PMID- 15121774 TI - Glutathione S-transferase M1, T1 status and the risk of head and neck cancer: a meta-analysis. PMID- 15121775 TI - Spondylo-epi-metaphyseal dysplasia (SEMD) matrilin 3 type: homozygote matrilin 3 mutation in a novel form of SEMD. PMID- 15121777 TI - PHOX2B mutations and polyalanine expansions correlate with the severity of the respiratory phenotype and associated symptoms in both congenital and late onset Central Hypoventilation syndrome. PMID- 15121778 TI - Embryonic expression of the human MID1 gene and its mutations in Opitz syndrome. PMID- 15121779 TI - Clinical and molecular analysis of Mowat-Wilson syndrome associated with ZFHX1B mutations and deletions at 2q22-q24.1. PMID- 15121780 TI - Zinc finger 81 (ZNF81) mutations associated with X-linked mental retardation. PMID- 15121782 TI - Refinement of the deletion in 7q21.3 associated with split hand/foot malformation type 1 and Mondini dysplasia. PMID- 15121783 TI - Genetic evidence for CFTR dysfunction in Japanese: background for chronic pancreatitis. PMID- 15121784 TI - FBN2 mutation associated with manifestations of Marfan syndrome and congenital contractural arachnodactyly. PMID- 15121785 TI - A variant form of hMTH1, a human homologue of the E coli mutT gene, correlates with somatic mutation in the p53 tumour suppressor gene in gastric cancer patients. PMID- 15121786 TI - Familial renal cell cancer appears to have a recessive component. PMID- 15121787 TI - Significant involvement of CCR2-64I and CXCL12-3a in the development of sporadic breast cancer. PMID- 15121788 TI - MDR1, the blood-brain barrier transporter, is associated with Parkinson's disease in ethnic Chinese. PMID- 15121789 TI - Mutations in the FKRP gene can cause muscle-eye-brain disease and Walker-Warburg syndrome. PMID- 15121790 TI - Modifier controls severity of a novel dominant low-frequency MyosinVIIA (MYO7A) auditory mutation. PMID- 15121791 TI - Evidence for association between single nucleotide polymorphisms in T complex protein 1 gene and schizophrenia in the Chinese Han population. PMID- 15121792 TI - Characterisation of a novel TSC2 missense mutation in the GAP related domain associated with minimal clinical manifestations of tuberous sclerosis. PMID- 15121793 TI - Molecular alterations in mitochondrial DNA of hepatocellular carcinomas: is there a correlation with clinicopathological profile? PMID- 15121794 TI - The common SCN5A mutation R1193Q causes LQTS-type electrophysiological alterations of the cardiac sodium channel. PMID- 15121795 TI - LMNA mutation in a 45 year old Japanese subject with Hutchinson-Gilford progeria syndrome. PMID- 15121796 TI - Clinical and molecular analysis of 30 patients with multiple lentigines LEOPARD syndrome. PMID- 15121797 TI - Somatic mosaicism is rare in unaffected parents of patients with sporadic tuberous sclerosis. PMID- 15121798 TI - Bayesian risk assessment for autosomal recessive diseases: fetal echogenic bowel with one or no detectable CFTR mutation. PMID- 15121799 TI - Maximal muscular vascular conductances during whole body upright exercise in humans. AB - That muscular blood flow may reach 2.5 l kg(-1) min(-1) in the quadriceps muscle has led to the suggestion that muscular vascular conductance must be restrained during whole body exercise to avoid hypotension. The main aim of this study was to determine the maximal arm and leg muscle vascular conductances (VC) during leg and arm exercise, to find out if the maximal muscular vasodilatory response is restrained during maximal combined arm and leg exercise. Six Swedish elite cross country skiers, age (mean +/-s.e.m.) 24 +/- 2 years, height 180 +/- 2 cm, weight 74 +/- 2 kg, and maximal oxygen uptake (VO(2,max)) 5.1 +/- 0.1 l min(-1) participated in the study. Femoral and subclavian vein blood flows, intra arterial blood pressure, cardiac output, as well as blood gases in the femoral and subclavian vein, right atrium and femoral artery were determined during skiing (roller skis) at approximately 76% of VO(2,max) and at VO(2,max) with different techniques: diagonal stride (combined arm and leg exercise), double poling (predominantly arm exercise) and leg skiing (predominantly leg exercise). During submaximal exercise cardiac output (26-27 l min(-1)), mean blood pressure (MAP) (approximately 87 mmHg), systemic VC, systemic oxygen delivery and pulmonary VO2(approximately 4 l min(-1)) attained similar values regardless of exercise mode. The distribution of cardiac output was modified depending on the musculature engaged in the exercise. There was a close relationship between VC and VO2 in arms (r= 0.99, P < 0.001) and legs (r= 0.98, P < 0.05). Peak arm VC (63.7 +/- 5.6 ml min(-1) mmHg(-1)) was attained during double poling, while peak leg VC was reached at maximal exercise with the diagonal technique (109.8 +/- 11.5 ml min(-1) mmHg(-1)) when arm VC was 38.8 +/- 5.7 ml min(-1) mmHg(-1). If during maximal exercise arms and legs had been vasodilated to the observed maximal levels then mean arterial pressure would have dropped at least to 75-77 mmHg in our experimental conditions. It is concluded that skeletal muscle vascular conductance is restrained during whole body exercise in the upright position to avoid hypotension. PMID- 15121800 TI - The effects of birth weight and postnatal growth patterns on fat depth and plasma leptin concentrations in juvenile and adult pigs. AB - Low birth weight is associated with altered adipose tissue deposition and regulation of leptin production. This study determined the effects of naturally occurring variations in birth weight in pigs on postnatal growth patterns, body fat depth and plasma leptin and other hormone concentrations. Low (< 1.47 kg) and high (> 1.53 kg) birth weight piglets were studied at 3 months (juvenile; n= 47) and 12 months of age (young adult; n= 17). At each age, arterial and venous catheters were inserted under general anaesthesia. Plasma leptin, cortisol, glucose, insulin and catecholamine concentrations were determined in basal blood samples. Body fat depth was measured by ultrasound at 12 months of age. Overall, adult fat depth was greater in low compared to high birth weight pigs and increased fat depth was associated with thinness at birth and poor early growth rates. These effects were strongest in females. Fat depth was related to current weight only in males. Compared to high birth weight pigs, plasma leptin concentrations were reduced in low birth weight females at 3 months and in low birth weight males at 12 months of age. This study demonstrates sex-specific effects of low birth weight on postnatal growth and body fatness and on plasma leptin concentrations in pigs. PMID- 15121802 TI - Changes in satellite cells in human skeletal muscle after a single bout of high intensity exercise. AB - No studies to date have reported activation of satellite cells in vivo in human muscle after a single bout of high intensity exercise. In this investigation, eight individuals performed a single bout of high intensity exercise with one leg, the contralateral leg being the control. A significant increase in mononuclear cells staining for the neural cell adhesion molecule (N-CAM) and fetal antigen 1 (FA1) were observed within the exercised human vastus lateralis muscle on days 4 and 8 post exercise. In addition, a significant increase in the concentration of the FA1 protein was determined in intramuscular dialysate samples taken from the vastus lateralis muscle of the exercising leg (day 0: 1.89 +/- 0.82 ng ml(-1); day 2: 1.68 +/- 0.37 ng ml(-1); day 4: 3.26 +/- 1.29 ng ml( 1), P < 0.05 versus basal; day 8: 4.68 +/- 2.06 ng ml(-1), P < 0.05 versus basal and control). No change was noted in the control leg. Despite this increase in N CAM- and FA1-positive mononuclear cells, an increased expression of myogenin and the neonatal isoform of the myosin heavy chain (MHCn) was not observed. Interestingly, myofibre lesions resulting from extensive damage to the proteins within the myofibre, particularly desmin or dystrophin, were not observed, and hence did not appear to induce the expression of either N-CAM or FA1. We therefore propose that satellite cells can be induced to re-enter the cell growth cycle after a single bout of unaccustomed high intensity exercise. However, a single bout of exercise is not sufficient for the satellite cell to undergo terminal differentiation. PMID- 15121801 TI - Utilization of triacylglycerol-rich lipoproteins by the working rat heart: routes of uptake and metabolic fates. AB - Very-low-density lipoprotein (VLDL) and chylomicrons (CMs) transport triacylglycerol (TAG) to peripheral tissues. Lipoprotein-TAG may gain access to target cells by lipoprotein lipase (LPL) hydrolysis or via receptor-mediated uptake; the principal routes of entry of VLDL and CM into heart are unknown, and different routes of entry may result in different metabolic fates. To examine this, isolated working rat hearts were perfused with rat VLDL and CMs, dual labelled with [3H]TAG and [14C]cholesterol. Uptake and utilization of CM-TAG were significantly greater than VLDL-TAG, but both were decreased significantly (more than halved) by tetrahydrolipstatin (THL, an inhibitor of lipoprotein lipase). By contrast, uptake of VLDL-cholesterol was much higher than CM-cholesterol (P < 0.01), and suramin (a lipoprotein receptor antagonist) decreased cholesterol uptake of both forms. CM-TAG oxidation rate was more than 4-fold higher than VLDL TAG oxidation. However, suramin decreased TAG oxidation from both VLDL and CM without affecting TAG uptake or total utilization, suggesting that the TAG gaining access through receptor-mediated pathways is preferentially 'channelled' towards oxidation. Most (79%) CM-TAG was oxidized whilst the proportion of VLDL TAG oxidized was only about half (49%). In the presence of suramin, there was a significant increase in esterification (incorporation of assimilated [3H]TAG into myocardial tissue [3H]lipids, mainly TAG) of assimilated TAG from both VLDL and CMs, again suggesting that receptor-mediated TAG uptake is directed towards oxidation rather than esterification. The importance of this relatively small pool of TAG is indicated by the fact that cardiac mechanical function declined markedly when lipoprotein receptors were inhibited. These results suggest that CMs, most fatty acids of which gain access into cardiomyocytes through LPL mediated hydrolysis, are the major supplier of TAG for hearts to oxidize; however, the metabolic fate of VLDL was split evenly between oxidation and deposition as myocardial tissue lipid. Most importantly, VLDL may play a regulatory role in heart lipid metabolism through a lipoprotein receptor-mediated mechanism. PMID- 15121803 TI - Functional characterization of a Ca(2+)-activated non-selective cation channel in human atrial cardiomyocytes. AB - Cardiac arrhythmias, which occur in a wide variety of conditions where intracellular calcium is increased, have been attributed to the activation of a transient inward current (Iti). Iti is the result of three different [Ca]i sensitive currents: the Na(+)-Ca2+ exchange current, a Ca(2+)-activated chloride current and a Ca(2+)-activated non-selective cationic current. Using the cell free configuration of the patch-clamp technique, we have characterized the properties of a Ca(2+)-activated non-selective cation channel (NSC(Ca)) in freshly dissociated human atrial cardiomyocytes. In excised inside-out patches, the channel presented a linear I-V relationship with a conductance of 19 +/- 0.4 pS. It discriminated poorly among monovalent cations (Na+ and K+) and was slightly permeable to Ca2+ ions. The channel's open probability was increased by depolarization and a rise in internal calcium, for which the Kd for [Ca2+]i was 20.8 microM. Channel activity was reduced in the presence of 0.5 mM ATP or 10 microM glibenclamide on the cytoplasmic side to 22.1 +/- 16.8 and 28.5 +/- 8.6%, respectively, of control. It was also inhibited by 0.1 mM flufenamic acid. The channel shares several properties with TRPM4b and TRPM5, two members of the 'TRP melastatin' subfamily. In conclusion, the NSC(Ca) channel is a serious candidate to support the delayed after-depolarizations observed in [Ca2+] overload and thus may be implicated in the genesis of arrhythmias. PMID- 15121804 TI - The excitability of lumbar motoneurones in the neonatal rat is increased by a hyperpolarization of their voltage threshold for activation by descending serotonergic fibres. AB - Previous work has shown there is an increase in motoneurone excitability produced by hyperpolarization of the threshold potential at which an action potential is elicited (Vth) at the onset, and throughout brainstem-induced fictive locomotion in the decerebrate cat. This represents a transient facilitation in the membrane potential for activation dependent on the presence of fictive locomotion. The present study tests the hypothesis that a similar neuromodulatory mechanism facilitating neuronal recruitment also exists in the neonatal rat, and the endogenous pathway mediating the Vth hyperpolarization can be activated by electrical stimulation of the neonatal brainstem. Isolated brainstem-spinal cord preparations from 1- to 5-day-old neonatal rats, and whole-cell recording techniques were used to examine the patterns of ventral root (VR) activity produced, and the effect of electrical stimulation of the ventromedial medulla on lumbar spinal neurones. Hyperpolarization of Vth was seen in 10/11 (range -2 to 18 mV) neurones recorded during locomotor-like VR activity, and appeared analogous to the locomotor-dependent Vth hyperpolarization previously described in the cat. However, in the present study, Vth hyperpolarization was also seen during electrical brainstem stimulation that evoked alternating, rhythmic, or tonic VR activity, or failed to evoke VR activity. Thirty-six of 71 neurones were antidromically identified as lumbar motoneurones and 33/36 showed a hyperpolarization of Vth (-2 to -14 mV) during electrical brainstem stimulation. Of the unidentified lumbar ventral horn neurones, 31/35 also showed hyperpolarization of Vth (-2 to -20 mV) during brainstem stimulation. The hyperpolarization of Vth and VR activity induced by brainstem stimulation was reversibly blocked by cooling of the cervical cord, indicating it is mediated by descending fibres, and application of the serotonergic antagonist ketanserin to the spinal cord was effectively able to block the brainstem-evoked hyperpolarization of Vth. These results demonstrate a previously unknown action of the endogenous descending serotonergic system to facilitate spinal motoneuronal recruitment and firing by inducing a hyperpolarization of Vth. This modulatory process can be examined in the neonatal rat brainstem-spinal cord preparation without the requirement for ongoing locomotor activity. PMID- 15121807 TI - Henry Head and his paradoxical reflex. PMID- 15121805 TI - Haemodynamic profile and responsiveness to anandamide of TRPV1 receptor knock-out mice. AB - The endocannabinoid anandamide and cannabinoid (CB) receptors have been implicated in the hypotension in various forms of shock and in advanced liver cirrhosis. Anandamide also activates vanilloid TRPV(1) receptors on sensory nerve terminals, triggering the release of calcitonin gene-related peptide which elicits vasorelaxation in isolated blood vessels in vitro. However, the contribution of TRPV(1) receptors to the in vivo hypotensive effect of anandamide is equivocal. We compared the cardiac performance of anaesthetized TRPV(1) knockout (TRPV(1)(-/-)) mice and their wild-type (TRPV(1)(+/+)) littermates and analysed in detail the haemodynamic effects of anandamide using the Millar pressure-volume conductance catheter system. Baseline cardiovascular parameters and systolic and diastolic function at different preloads were similar in TRPV(1)(-/-) and TRPV(1)(+/+) mice. The predominant hypotensive response to bolus intravenous injections of anandamide and the associated decrease in cardiac contractility and total peripheral resistance (TPR) were similar in TRPV(1)(+/+) and TRPV(1)(-/-) mice, as was the ability of the CB(1) receptor antagonist SR141716 to completely block these effects. In TRPV(1)(+/+) mice, this hypotensive response was preceded by a transient, profound drop in cardiac contractility and heart rate and an increase in TPR, followed by a brief pressor response, effects which were unaffected by SR141716 and were absent in TRPV(1)(-/ ) mice. These results indicate that mice lacking TRPV(1) receptors have a normal cardiovascular profile and their predominant cardiovascular depressor response to anandamide is mediated through CB(1) receptors. The role of TRPV(1) receptors is limited to the transient activation of the Bezold-Jarisch reflex by very high initial plasma concentrations of anandamide. PMID- 15121808 TI - Newborn screening for cystic fibrosis. PMID- 15121806 TI - A role for SNAP-25 but not VAMPs in store-mediated Ca2+ entry in human platelets. AB - Store-mediated Ca2+ entry (SMCE) is a major mechanism for Ca2+ influx in non excitable cells. Recently, a conformational coupling mechanism allowing coupling between transient receptor potential channels (TRPCs) and IP3 receptors has been proposed to activate SMCE. Here we have investigated the role of two soluble N ethylmaleimide-sensitive-factor attachment protein receptors (SNAREs), which are involved in membrane trafficking and docking, in SMCE in human platelets. We found that the synaptosome-associated protein (SNAP-25) and the vesicle associated membrane proteins (VAMP) coimmunoprecipitate with hTRPC1 in platelets. Treatment with botulinum toxin (BoNT) E or with tetanus toxin (TeTx), induced cleavage and inactivation of SNAP-25 and VAMPs, respectively. BoNTs significantly reduced thapsigargin- (TG) and agonist-evoked SMCE. Treatment with BoNTs once SMCE had been activated decreased Ca2+ entry, indicating that SNAP-25 is required for the activation and maintenance of SMCE. In contrast, treatment with TeTx had no effect on either the activation or the maintenance of SMCE in platelets. Finally, treatment with BoNT E impaired the coupling between naturally expressed hTRPC1 and IP3 receptor type II in platelets. From these findings we suggest SNAP 25 has a role in SMCE in human platelets. PMID- 15121809 TI - Regulation of the medical profession: fantasy, reality and legality. PMID- 15121810 TI - Medically unexplained symptoms: the biopsychosocial model found wanting. PMID- 15121811 TI - Diagnosis and treatment of hepatitis C. PMID- 15121812 TI - Clinical management of children and young adults with heterozygous familial hypercholesterolaemia in the UK. AB - Life expectancy in familial hypercholesterolaemia (FH) has been greatly improved by the advent of statin therapy. In the UK, however, these agents are not licensed for use in children. We approached 169 physicians responsible for lipid clinics for information on their practice in young patients, and valid responses were received from 54%. A typical lipid clinic has only 3.5 patients aged under 16 with FH. In boys aged 10-15 years 65% of physicians were prepared to treat with bile acid sequestrants but only 23% with statins. There was greater reluctance to treat in girls of the same age, corresponding figures being 52% and 12%. Despite the efficacy of statins in reducing low-density-lipoprotein cholesterol, these agents are little used in children with FH. Their safety and clinical efficacy should be assessed by a randomized double-blind trial. PMID- 15121814 TI - Hospital bathrooms and showers: a continuing saga of inadequacy. AB - Previous surveys of UK hospitals have highlighted many deficiencies in the standards of hospital inpatient washing and bathing facilities--especially inadequate access for wheelchair users, insufficient bathing equipment, and unsatisfactory cleanliness and privacy. We conducted a qualitative survey in three hospitals in the North of England to see whether these facilities have improved. There have been some improvements, particularly in the provision of bath hoists, adapted taps, alarm call systems, shower seats and wheelchair access to bathrooms. But many basic problems remain-absent locks and signs, inadequate heating, poor standards of privacy, insufficient bath aids, wet floors, and the inappropriate use of bathrooms as store rooms. The overall condition of hospital bathrooms and showers remains unsatisfactory. Too many hospital bathrooms are austere, cold, smelly and poorly maintained. PMID- 15121813 TI - Prehospital thrombolysis--calculated health benefit for catchment population of one hospital. AB - The health benefit of thrombolysis in acute myocardial infarction is greatest when patients are treated soon after onset of symptoms. One approach to reducing treatment delay is to give thrombolysis before the patient reaches hospital. When an ambulance trust proposed a prehospital thrombolysis service, local commissioners requested an estimate of its possible health impact. Clinical audit and ambulance trust data were obtained for 165 patients who received thrombolysis for acute myocardial infarction in the coronary care unit of a local hospital in one year. This information was then used to estimate the health impact of prehospital thrombolysis in the local population in a mathematical model derived from the results of trials comparing prehospital and hospital thrombolysis. The best predicted local health benefit from the proposed prehospital thrombolysis service is that, if 45 minutes can be cut off the call-to-needle time, 61 cases of acute myocardial infarction need to be treated to save one additional life at 35 days. By use of published research data, the health benefits of prehospital thrombolysis can be estimated for a local population. Variables in the treatment population and ambulance service will influence the size of the health benefit that can be achieved. PMID- 15121815 TI - Vitamin D deficiency and cardiac failure in infancy. PMID- 15121816 TI - Beware of the midline scalp lump. PMID- 15121817 TI - Gastric ligneoma. PMID- 15121818 TI - French without tears? Foreign accent syndrome. PMID- 15121819 TI - The Bradford smallpox outbreak in 1962: a personal account. PMID- 15121821 TI - Swallowed partial dentures. PMID- 15121822 TI - Swallowed partial dentures. PMID- 15121823 TI - Obesity in Chinese children. PMID- 15121824 TI - Shell shock. PMID- 15121825 TI - Swallowed toothbrush. PMID- 15121826 TI - Putrid gums and 'dead men's cloaths'. PMID- 15121827 TI - An unusual paracetamol overdose. PMID- 15121828 TI - Laennec's legacy. PMID- 15121829 TI - Role of thyroxine in chronic urticaria and angio-oedema. PMID- 15121830 TI - Risk factors for neonaticide and infant homicide. PMID- 15121831 TI - Key role of Ser562/661 in Snf1-dependent regulation of Cat8p in Saccharomyces cerevisiae and Kluyveromyces lactis. AB - Utilization of nonfermentable carbon sources by Kluyveromyces lactis and Saccharomyces cerevisiae requires the Snf1p kinase and the Cat8p transcriptional activator, which binds to carbon source-responsive elements of target genes. We demonstrate that KlSnf1p and KlCat8p from K. lactis interact in a two-hybrid system and that the interaction is stronger with a kinase-dead mutant form of KlSnf1p. Of two putative phosphorylation sites in the KlCat8p sequence, serine 661 was identified as a key residue governing KlCat8p regulation. Serine 661 is located in the middle homology region, a regulatory domain conserved among zinc cluster transcription factors, and is part of an Snf1p consensus phosphorylation site. Single mutations at this site are sufficient to completely change the carbon source regulation of the KlCat8p transactivation activity observed. A serine-to-glutamate mutant form mimicking constitutive phosphorylation results in a nearly constitutively active form of KlCat8p, while a serine-to-alanine mutation has the reverse effect. Furthermore, it is shown that KlCat8p phosphorylation depends on KlSNF1. The Snf1-Cat8 connection is evolutionarily conserved: mutation of corresponding serine 562 of ScCat8p gave similar results in S. cerevisiae. The enhanced capacity of ScCat8S562E to suppress the phenotype caused by snf1 strengthens the hypothesis of direct phosphorylation of Cat8p by Snf1p. Unlike that of S. cerevisiae ScCAT8, KlCAT8 transcription is not carbon source regulated, illustrating the prominent role of posttranscriptional regulation of Cat8p in K. lactis. PMID- 15121832 TI - The two steps of poly(A)-dependent termination, pausing and release, can be uncoupled by truncation of the RNA polymerase II carboxyl-terminal repeat domain. AB - The carboxyl-terminal repeat domain (CTD) of RNA polymerase II is thought to help coordinate events during RNA metabolism. The mammalian CTD consists of 52 imperfectly repeated heptads followed by 10 additional residues at the C terminus. The CTD is required for cleavage and polyadenylation in vitro. We studied poly(A)-dependent termination in vivo using CTD truncation mutants. Poly(A)-dependent termination occurs in two steps, pause and release. We found that the CTD is required for release, the first 25 heptads being sufficient. Neither the final 10 amino acids nor the variant heptads of the second half of the CTD were required. No part of the CTD was required for poly(A)-dependent pausing--the poly(A) signal could communicate directly with the body of the polymerase. By removing the CTD, pausing could be observed without being obscured by release. Poly(A)-dependent pausing appeared to operate by slowing down the polymerase, such as by down-regulation of a positive elongation factor. Although the first 25 heptads supported undiminished poly(A)-dependent termination, they did not efficiently support events near the promoter involved in abortive elongation. However, the second half of the CTD, including the final 10 amino acids, was sufficient for these functions. PMID- 15121833 TI - An array of coactivators is required for optimal recruitment of TATA binding protein and RNA polymerase II by promoter-bound Gcn4p. AB - Wild-type transcriptional activation by Gcn4p is dependent on multiple coactivators, including SAGA, SWI/SNF, Srb mediator, CCR4-NOT, and RSC, which are all recruited by Gcn4p to its target promoters in vivo. It was not known whether these coactivators are required for assembly of the preinitiation complex (PIC) or for subsequent steps in the initiation or elongation phase of transcription. We find that mutations in subunits of these coactivators reduce the recruitment of TATA binding protein (TBP) and RNA polymerase II (Pol II) by Gcn4p at ARG1, ARG4, and SNZ1, implicating all five coactivators in PIC assembly at Gcn4p target genes. Recruitment of Pol II at SNZ1 and ARG1 was eliminated by mutations in TBP or by deletion of the TATA box, indicating that TBP binding is a prerequisite for Pol II recruitment by Gcn4p. However, several mutations in SAGA subunits and deletion of SRB10 had a greater impact on promoter occupancy of Pol II versus TBP, suggesting that SAGA and Srb mediator can promote Pol II binding independently of their stimulatory effects on TBP recruitment. Our results reveal an unexpected complexity in the cofactor requirements for the enhancement of PIC assembly by a single activator protein. PMID- 15121834 TI - Genome-wide occupancy profile of the RNA polymerase III machinery in Saccharomyces cerevisiae reveals loci with incomplete transcription complexes. AB - We used chromatin immunoprecipitation, followed by microarray hybridization, to determine the genome-wide distribution of the RNA polymerase (Pol) III transcription apparatus in the yeast Saccharomyces cerevisiae. The Pol III transcriptome includes all tRNA genes, previously identified non-tRNA Pol III genes, and SNR52, which encodes a small nucleolar RNA. Unexpectedly, we identify eight ETC loci that are occupied by TFIIIC but not by other components of the Pol III machinery. Some ETC loci contain stretches of DNA that are highly conserved among closely related yeast species, suggesting that they may encode functional RNAs. ETC6 is located upstream of the gene encoding the tau 91 subunit of TFIIIC, suggesting the possibility of Pol III-regulated expression of a critical Pol III factor. We also identify the ZOD1 locus, which is bound by all components of the Pol III machinery and yet does not appear to express an RNA conserved among closely related yeast species. The B block motifs and several flanking nucleotides of the ZOD1 and ETC loci are very similar to each other and are highly conserved across the yeast species. Furthermore, the unusual profile of Pol III factor association with ZOD1 and the ETC loci is perfectly preserved in a different Saccharomyces species, indicating that these loci represent novel functional entities. PMID- 15121835 TI - Glucose utilization is essential for hypoxia-inducible factor 1 alpha-dependent phosphorylation of c-Jun. AB - Hypoxia and anoxia are important microenvironmental stresses that contribute to pathological events such as solid-tumor development. We have been investigating the effects of hypoxia and anoxia on expression of the proto-oncogene c-jun and the regulation of c-Jun/AP-1 transcription factors. In earlier work using genetically manipulated mouse embryo fibroblasts (mEFs), we found a functional relationship among c-jun expression, c-Jun N-terminal phosphorylation, and the presence of hypoxia-inducible factor 1 alpha (HIF-1 alpha), the oxygen-regulated subunit of the HIF-1 transcription factor. Both the induction of c-jun mRNA expression and c-Jun N-terminal phosphorylation in cells exposed to hypoxia or anoxia were found to be dependent on the presence of HIF-1 alpha, but this was not the case in cells exposed to less-severe hypoxia. Here we describe new findings concerning HIF-1-dependent c-Jun N-terminal phosphorylation in cells exposed to hypoxia or anoxia. Specifically, we report that hypoxia-inducible c Jun N-terminal kinase (JNK) activity, which involves JNKs or stress-activated protein kinases (SAPKs), is dependent on enhanced glucose utilization mediated by HIF-1. These results suggest a model in which hypoxia-inducible JNK activity is connected to oxygen sensing through increased glucose absorption and/or glycolytic activity regulated by the HIF-1 system. We also found that basal threonine and tyrosine phosphorylation (within the TEY motif) of extracellular signal-regulated kinases 1 and 2 (ERK1/2) and the corresponding ERK1/2 activity were defective in hypoxic HIF-1 alpha-null mEFs but not in wild-type mEFs, independently of glucose uptake. Therefore, the activities of both JNKs/SAPKs and ERK1/2 are sensitive to HIF-1-dependent processes in cells exposed to hypoxia or anoxia. PMID- 15121836 TI - Defined sequence modules and an architectural element cooperate to promote initiation at an ectopic mammalian chromosomal replication origin. AB - A small DNA fragment containing the high-frequency initiation region (IR) ori beta from the hamster dihydrofolate reductase locus functions as an independent replicator in ectopic locations in both hamster and human cells. Conversely, a fragment of the human lamin B2 locus containing the previously mapped IR serves as an independent replicator at ectopic chromosomal sites in hamster cells. At least four defined sequence elements are specifically required for full activity of ectopic ori-beta in hamster cells. These include an AT-rich element, a 4-bp sequence located within the mapped IR, a region of intrinsically bent DNA located between these two elements, and a RIP60 protein binding site adjacent to the bent region. The ori-beta AT-rich element is critical for initiation activity in human, as well as hamster, cells and can be functionally substituted for by an AT rich region from the human lamin B2 IR that differs in nucleotide sequence and length. Taken together, the results demonstrate that two mammalian replicators can be activated at ectopic sites in chromosomes of another mammal and lead us to speculate that they may share functionally related elements. PMID- 15121838 TI - Hypertension and impaired glycine handling in mice lacking the orphan transporter XT2. AB - A family of orphan transporters has been discovered that are structurally related to the Na(+)-Cl(-)-dependent neurotransmitter transporters, including the dopamine transporter. One member of this family, the mouse XT2 gene, is predominantly expressed in the kidney and has 95% homology to rat ROSIT (renal osmotic stress-induced Na(+)-Cl(-) organic solute cotransporter). To study the physiological functions of this transporter, we generated XT2-knockout mice by gene targeting. XT2(-/-) mice develop and survive normally with no apparent abnormalities. To attempt to identify potential substrates for XT2, we screened urine from XT2-knockout mice by high-pressure liquid chromatography and mass spectrometry and found significantly elevated concentrations of glycine. To study glycine handling, XT2(+/+) and XT2(-/-) mice were injected with radiolabeled glycine, and urine samples were collected to monitor glycine excretion. After 2 h, XT2(-/-) mice were found to excrete almost twice as much glycine as the XT2(+/+) controls (P = 0.03). To determine whether the absence of the XT2 transporter affected sodium and fluid homeostasis, we measured systolic blood pressure by computerized tail-cuff manometry. Systolic blood pressure was significantly higher in XT2(-/-) mice (127 +/- 3 mmHg) than in wild-type controls (114 +/- 2 mmHg; P < 0.001). This difference in systolic blood pressure was maintained on high and low salt feeding. To examine whether the alteration in blood pressure and the defect in glycine handling were related, we measured systolic blood pressure in the XT2(-/-) mice during dietary glycine supplementation. Glycine loading caused systolic blood pressure to fall in the XT2(-/-) mice from 127 +/- 3 to 115 +/- 3 mmHg (P < 0.001), a level virtually identical to that of the wild-type controls. These data suggest that the XT2 orphan transporter is involved in glycine reabsorption and that the absence of this transporter is sufficient to cause hypertension. PMID- 15121837 TI - The functions of budding yeast Sae2 in the DNA damage response require Mec1- and Tel1-dependent phosphorylation. AB - DNA damage checkpoint pathways sense DNA lesions and transduce the signals into appropriate biological responses, including cell cycle arrest, induction of transcriptional programs, and modification or activation of repair factors. Here we show that the Saccharomyces cerevisiae Sae2 protein, known to be involved in processing meiotic and mitotic double-strand breaks, is required for proper recovery from checkpoint-mediated cell cycle arrest after DNA damage and is phosphorylated periodically during the unperturbed cell cycle and in response to DNA damage. Both cell cycle- and DNA damage-dependent Sae2 phosphorylation requires the main checkpoint kinase, Mec1, and the upstream components of its pathway, Ddc1, Rad17, Rad24, and Mec3. Another pathway, involving Tel1 and the MRX complex, is also required for full DNA damage-induced Sae2 phosphorylation, that is instead independent of the downstream checkpoint transducers Rad53 and Chk1, as well as of their mediators Rad9 and Mrc1. Mutations altering all the favored ATM/ATR phosphorylation sites of Sae2 not only abolish its in vivo phosphorylation after DNA damage but also cause hypersensitivity to methyl methanesulfonate treatment, synthetic lethality with RAD27 deletion, and decreased rates of mitotic recombination between inverted Alu repeats, suggesting that checkpoint-mediated phosphorylation of Sae2 is important to support its repair and recombination functions. PMID- 15121839 TI - Polypyrimidine tract binding protein modulates efficiency of polyadenylation. AB - Polypyrimidine tract binding protein (PTB) is a major hnRNP protein with multiple roles in mRNA metabolism, including regulation of alternative splicing and internal ribosome entry site-driven translation. We show here that a fourfold overexpression of PTB results in a 75% reduction of mRNA levels produced from transfected gene constructs with different polyadenylation signals (pA signals). This effect is due to the reduced efficiency of mRNA 3' end cleavage, and in vitro analysis reveals that PTB competes with CstF for recognition of the pA signal's pyrimidine-rich downstream sequence element. This may be analogous to its role in alternative splicing, where PTB competes with U2AF for binding to pyrimidine-rich intronic sequences. The pA signal of the C2 complement gene unusually possesses a PTB-dependent upstream sequence, so that knockdown of PTB expression by RNA interference reduces C2 mRNA expression even though PTB overexpression still inhibits polyadenylation. Consequently, we show that PTB can act as a regulator of mRNA expression through both its negative and positive effects on mRNA 3' end processing. PMID- 15121840 TI - A conserved docking motif for CK1 binding controls the nuclear localization of NFAT1. AB - In resting cells, the NFAT1 transcription factor is kept inactive in the cytoplasm by phosphorylation on multiple serine residues. These phosphorylated residues are primarily contained within two types of serine-rich motifs, the SRR 1 and SP motifs, which are conserved within the NFAT family. Several different kinases have been proposed to regulate NFAT, but no single candidate displays the specificity required to fully phosphorylate both types of motifs; thus, the identity of the kinase that regulates NFAT activity remains unclear. Here we show that the NFAT1 serine motifs are regulated by distinct kinases that must coordinate to control NFAT1 activation. CK1 phosphorylates only the SRR-1 motif, the primary region required for NFAT1 nuclear import. CK1 exists with NFAT1 in a high-molecular-weight complex in resting T cells but dissociates upon activation. GSK3 does not phosphorylate the SRR-1 region but can target the NFAT1 SP-2 motif, and it synergizes with CK1 to regulate NFAT1 nuclear export. We identify a conserved docking site for CK1 in NFAT proteins and show that mutation of this site disrupts NFAT1-CK1 interaction and causes constitutive nuclear localization of NFAT1. The CK1 docking motif is present in proteins of the Wnt, Hedgehog, and circadian-rhythm pathways, which also integrate the activities of CK1 and GSK3. PMID- 15121841 TI - Identification of factors regulating poly(A) tail synthesis and maturation. AB - Posttranscriptional maturation of the 3' end of eukaryotic pre-mRNAs occurs as a three-step pathway involving site-specific cleavage, polymerization of a poly(A) tail, and trimming of the newly synthesized tail to its mature length. While most of the factors essential for catalyzing these reactions have been identified, those that regulate them remain to be characterized. Previously, we demonstrated that the yeast protein Pbp1p associates with poly(A)-binding protein (Pab1p) and controls the extent of mRNA polyadenylation. To further elucidate the function of Pbp1p, we conducted a two-hybrid screen to identify factors with which it interacts. Five genes encoding putative Pbp1p-interacting proteins were identified, including (i) FIR1/PIP1 and UFD1/PIP3, genes encoding factors previously implicated in mRNA 3'-end processing; (ii) PBP1 itself, confirming directed two-hybrid results and suggesting that Pbp1p can multimerize; (iii) DIG1, encoding a mitogen-activated protein kinase-associated protein; and (iv) PBP4 (YDL053C), a previously uncharacterized gene. In vitro polyadenylation reactions utilizing extracts derived from fir1 Delta and pbp1 Delta cells and from cells lacking the Fir1p interactor, Ref2p, demonstrated that Pbp1p, Fir1p, and Ref2p are all required for the formation of a normal-length poly(A) tail on precleaved CYC1 pre-mRNA. Kinetic analyses of the respective polyadenylation reactions indicated that Pbp1p is a negative regulator of poly(A) nuclease (PAN) activity and that Fir1p and Ref2p are, respectively, a positive regulator and a negative regulator of poly(A) synthesis. We suggest a model in which these three factors and Ufd1p are part of a regulatory complex that exploits Pab1p to link cleavage and polyadenylation factors of CFIA and CFIB (cleavage factors IA and IB) to the polyadenylation factors of CPF (cleavage and polyadenylation factor). PMID- 15121842 TI - The Sox-2 regulatory regions display their activities in two distinct types of multipotent stem cells. AB - The Sox-2 gene is expressed in embryonic stem (ES) cells and neural stem cells. Two transcription enhancer regions, Sox-2 regulatory region 1 (SRR1) and SRR2, were described previously based on their activities in ES cells. Here, we demonstrate that these regulatory regions also exert their activities in neural stem cells. Moreover, our data reveal that, as in ES cells, both SRR1 and SRR2 show their activities rather specifically in multipotent neural stem or progenitor cells but cease to function in differentiated cells, such as postmitotic neurons. Systematic deletion and mutation analyses showed that the same or at least overlapping DNA elements of SRR2 are involved in its activity in both ES and neural stem or progenitor cells. Thus, SRR2 is the first example of an enhancer in which a single regulatory core sequence is involved in multipotent state-specific expression in two different stem cells, i.e., ES and neural stem cells. PMID- 15121843 TI - Normal embryonic and germ cell development in mice lacking alpha 1,3 fucosyltransferase IX (Fut9) which show disappearance of stage-specific embryonic antigen 1. AB - Stage-specific embryonic antigen 1 (SSEA-1), an antigenic epitope defined as a Lewis x carbohydrate structure, is expressed during the 8-cell to blastocyst stages in mouse embryos and in primordial germ cells, undifferentiated embryonic stem cells, and embryonic carcinoma cells. For many years, SSEA-1 has been implicated in the development of mouse embryos as a functional carbohydrate epitope in cell-to-cell interaction during morula compaction. In a previous study, alpha 1,3-fucosyltransferase IX (Fut9) exhibited very strong activity for the synthesis of Lewis x compared to other alpha 1,3-fucosyltransferases in an in vitro substrate specificity assay. Fut4 and Fut9 transcripts were expressed in mouse embryos. The Fut9 transcript was detected in embryonic-day-13.5 gonads containing primordial germ cells, but the Fut4 transcript was not. In order to identify the role of SSEA-1 and determine the key enzyme for SSEA-1 synthesis in vivo, we have generated Fut9-deficient (Fut9(-/-)) mice. Fut9(-/-) mice develop normally, with no gross phenotypic abnormalities, and are fertile. Immunohistochemical analysis revealed an absence of SSEA-1 expression in early embryos and primordial germ cells of Fut9(-/-) mice. Therefore, we conclude that expression of the SSEA-1 epitope in the developing mouse embryo is not essential for embryogenesis in vivo. PMID- 15121844 TI - The splicing factor U2AF small subunit is functionally conserved between fission yeast and humans. AB - The small subunit of U2AF, which functions in 3' splice site recognition, is more highly conserved than its heterodimeric partner yet is less thoroughly investigated. Remarkably, we find that the small subunit of Schizosaccharomyces pombe U2AF (U2AF(SM)) can be replaced in vivo by its human counterpart, demonstrating that the conservation extends to function. Precursor mRNAs accumulate in S. pombe following U2AF(SM) depletion in a time frame consistent with a role in splicing. A comprehensive mutational analysis reveals that all three conserved domains are required for viability. Notably, however, a tryptophan in the pseudo-RNA recognition motif implicated in a key contact with the large subunit by crystallographic data is dispensable whereas amino acids implicated in RNA recognition are critical. Mutagenesis of the two zinc-binding domains demonstrates that they are neither equivalent nor redundant. Finally, two and three-hybrid analyses indicate that mutations with effects on large-subunit interactions are rare whereas virtually all alleles tested diminished RNA binding by the heterodimer. In addition to demonstrating extraordinary conservation of U2AF small-subunit function, these results provide new insights into the roles of individual domains and residues. PMID- 15121845 TI - Id2 and Id3 define the potency of cell proliferation and differentiation responses to transforming growth factor beta and bone morphogenetic protein. AB - Transforming growth factors beta (TGF-betas) inhibit growth of epithelial cells and induce differentiation changes, such as epithelial-mesenchymal transition (EMT). On the other hand, bone morphogenetic proteins (BMPs) weakly affect epithelial cell growth and do not induce EMT. Smad4 transmits signals from both TGF-beta and BMP pathways. Stimulation of Smad4-deficient epithelial cells with TGF-beta 1 or BMP-7 in the absence or presence of exogenous Smad4, followed by cDNA microarray analysis, revealed 173 mostly Smad4-dependent, TGF-beta-, or BMP responsive genes. Among 25 genes coregulated by both factors, inhibitors of differentiation Id2 and Id3 showed long-term repression by TGF-beta and sustained induction by BMP. The opposing regulation of Id genes is critical for proliferative and differentiation responses. Hence, ectopic Id2 or Id3 expression renders epithelial cells refractory to growth inhibition and EMT induced by TGF beta, phenocopying the BMP response. Knockdown of endogenous Id2 or Id3 sensitizes epithelial cells to BMP, leading to robust growth inhibition and induction of transdifferentiation. Thus, Id genes sense Smad signals and create a permissive or refractory nuclear environment that defines decisions of cell fate and proliferation. PMID- 15121846 TI - Characterization of mouse Rsk4 as an inhibitor of fibroblast growth factor-RAS extracellular signal-regulated kinase signaling. AB - Receptor tyrosine kinase (RTK) signals regulate the specification of a varied array of tissue types by utilizing distinct modules of proteins to elicit diverse effects. The RSK proteins are part of the RTK signal transduction pathway and are thought to relay these signals by acting downstream of extracellular signal regulated kinase (ERK). In this study we report the identification of ribosomal S6 kinase 4 (Rsk4) as an inhibitor of RTK signals. Among the RSK proteins, RTK inhibition is specific to RSK4 and, in accordance, is dependent upon a region of the RSK4 protein that is divergent from other RSK family members. We demonstrate that Rsk4 inhibits the transcriptional activation of specific targets of RTK signaling as well as the activation of ERK. Developmentally, Rsk4 is expressed in extraembryonic tissue, where RTK signals are known to have critical roles. Further examination of Rsk4 expression in the extraembryonic tissues demonstrates that its expression is inversely correlated with the presence of activated ERK 1/2. These studies demonstrate a new and divergent function for RSK4 and support a role for RSK proteins in the specification of RTK signals during early mouse development. PMID- 15121847 TI - Opposing effects of ubiquitin conjugation and SUMO modification of PCNA on replicational bypass of DNA lesions in Saccharomyces cerevisiae. AB - The Rad6-Rad18 ubiquitin-conjugating enzyme complex of Saccharomyces cerevisiae promotes replication through DNA lesions via three separate pathways that include translesion synthesis (TLS) by DNA polymerases zeta (Polzeta) and Poleta and postreplicational repair mediated by the Mms2-Ubc13 ubiquitin-conjugating enzyme and Rad5. Here we report our studies with a proliferating cell nuclear antigen (PCNA) mutation, pol30-119, which results from a change of the lysine 164 residue to arginine. It has been shown recently that following treatment of yeast cells with DNA-damaging agents, the lysine 164 residue of PCNA becomes monoubiquitinated in a Rad6-Rad18-dependent manner and that subsequently this PCNA residue is polyubiquitinated via a lysine 63-linked ubiquitin chain in an Mms2-Ubc13-, Rad5-dependent manner. PCNA is also modified by SUMO conjugation at the lysine 164 residue. Our genetic studies with the pol30-119 mutation show that in addition to conferring a defect in Polzeta-dependent UV mutagenesis and in Poleta-dependent TLS, this PCNA mutation inhibits postreplicational repair of discontinuities that form in the newly synthesized strand across from UV lesions. In addition, we provide evidence for the activation of the RAD52 recombinational pathway in the pol30-119 mutant and we infer that SUMO conjugation at the lysine 164 residue of PCNA has a role in suppressing the Rad52-dependent postreplicational repair pathway. PMID- 15121848 TI - Constitutive expression of tert in thymocytes leads to increased incidence and dissemination of T-cell lymphoma in Lck-Tert mice. AB - Here we describe a new mouse model with constitutive expression of the catalytic subunit of telomerase (Tert) targeted to thymocytes and peripheral T cells (Lck Tert mice). Two independent Lck-Tert mouse lines showed higher incidences of spontaneous T-cell lymphoma than the corresponding age-matched wild-type controls, indicating that constitutive expression of Tert promotes lymphoma. Interestingly, T-cell lymphomas in Lck-Tert mice were more disseminated than those in wild-type controls and affected both lymphoid and nonlymphoid tissues, while nonlymphoid tissues were never affected with lymphoma in age-matched wild type controls. Importantly, these roles of Tert constitutive expression in promoting tumor progression and dissemination were independent of the role of telomerase in telomere length maintenance, since telomere length distributions on a single-cell basis were identical in Lck-Tert and wild-type thymocytes. Finally, Tert constitutive expression did not interfere with telomere capping in Lck-Tert primary thymocytes, although it resulted in greater chromosomal instability upon gamma irradiation in Lck-Tert primary lymphocytes than in controls, suggesting that Tert overexpression may interfere with the cellular response to DNA damage. PMID- 15121849 TI - Visualization of Myc/Max/Mad family dimers and the competition for dimerization in living cells. AB - Myc and Mad family proteins play opposing roles in the control of cell growth and proliferation. We have visualized the subcellular locations of complexes formed by Myc/Max/Mad family proteins using bimolecular fluorescence complementation (BiFC) analysis. Max was recruited to different subnuclear locations by interactions with Myc versus Mad family members. Complexes formed by Max with Mxi1, Mad3, or Mad4 were enriched in nuclear foci, whereas complexes formed with Myc were more uniformly distributed in the nucleoplasm. Mad4 was localized to the cytoplasm when it was expressed separately, and Mad4 was recruited to the nucleus through dimerization with Max. The cytoplasmic localization of Mad4 was determined by a CRM1-dependent nuclear export signal located near the amino terminus. We compared the relative efficiencies of complex formation among Myc, Max, and Mad family proteins in living cells using multicolor BiFC analysis. Max formed heterodimers with the basic helix-loop-helix leucine zipper (bHLHZIP) domain of Myc (bMyc) more efficiently than it formed homodimers. Replacement of two amino acid residues in the leucine zipper of Max reversed the relative efficiencies of homo- and heterodimerization in cells. Surprisingly, Mad3 formed complexes with Max less efficiently than bMyc, whereas Mad4 formed complexes with Max more efficiently than bMyc. The distinct subcellular locations and the differences between the efficiencies of dimerization with Max indicate that Mad3 and Mad4 are likely to modulate transcription activation by Myc at least in part through distinct mechanisms. PMID- 15121850 TI - The Schizosaccharomyces pombe HIRA-like protein Hip1 is required for the periodic expression of histone genes and contributes to the function of complex centromeres. AB - HIRA-like (Hir) proteins are evolutionarily conserved and are implicated in the assembly of repressive chromatin. In Saccharomyces cerevisiae, Hir proteins contribute to the function of centromeres. However, S. cerevisiae has point centromeres that are structurally different from the complex centromeres of metazoans. In contrast, Schizosaccharomyces pombe has complex centromeres whose domain structure is conserved with that of human centromeres. Therefore, we examined the functions of the fission yeast Hir proteins Slm9 and the previously uncharacterised protein Hip1. Deletion of hip1(+) resulted in phenotypes that were similar to those described previously for slm9 Delta cells: a cell cycle delay, synthetic lethality with cdc25-22, and poor recovery from nitrogen starvation. However, while it has previously been shown that Slm9 is not required for the periodic expression of histone H2A, we found that loss of Hip1 led to derepression of core histone genes expression outside of S phase. Importantly, we found that deletion of either hip1(+) or slm9(+) resulted in increased rates of chromosome loss, increased sensitivity to spindle damage, and reduced transcriptional silencing in the outer centromeric repeats. Thus, S. pombe Hir proteins contribute to pericentromeric heterochromatin, and our data thus suggest that Hir proteins may be required for the function of metazoan centromeres. PMID- 15121851 TI - Network of dynamic interactions between histone H1 and high-mobility-group proteins in chromatin. AB - Histone H1 and the high-mobility group (HMG) proteins are chromatin binding proteins that regulate gene expression by modulating the compactness of the chromatin fiber and affecting the ability of regulatory factors to access their nucleosomal targets. Histone H1 stabilizes the higher-order chromatin structure and decreases nucleosomal access, while the HMG proteins decrease the compactness of the chromatin fiber and enhance the accessibility of chromatin targets to regulatory factors. Here we show that in living cells, each of the three families of HMG proteins weakens the binding of H1 to nucleosomes by dynamically competing for chromatin binding sites. The HMG families weaken H1 binding synergistically and do not compete among each other, suggesting that they affect distinct H1 binding sites. We suggest that a network of dynamic and competitive interactions involving HMG proteins and H1, and perhaps other structural proteins, constantly modulates nucleosome accessibility and the local structure of the chromatin fiber. PMID- 15121852 TI - Hip1-related mutant mice grow and develop normally but have accelerated spinal abnormalities and dwarfism in the absence of HIP1. AB - In mice and humans, there are two known members of the Huntingtin interacting protein 1 (HIP1) family, HIP1 and HIP1-related (HIP1r). Based on structural and functional data, these proteins participate in the clathrin trafficking network. The inactivation of Hip1 in mice leads to spinal, hematopoietic, and testicular defects. To investigate the biological function of HIP1r, we generated a Hip1r mutant allele in mice. Hip1r homozygous mutant mice are viable and fertile without obvious morphological abnormalities. In addition, embryonic fibroblasts derived from these mice do not have gross abnormalities in survival, proliferation, or clathrin trafficking pathways. Altogether, this demonstrates that HIP1r is not necessary for normal development of the embryo or for normal adulthood and suggests that HIP1 or other functionally related members of the clathrin trafficking network can compensate for HIP1r absence. To test the latter, we generated mice deficient in both HIP1 and HIP1r. These mice have accelerated development of abnormalities seen in Hip1 -deficient mice, including kypholordosis and growth defects. The severity of the Hip1r/Hip1 double-knockout phenotype compared to the Hip1 knockout indicates that HIP1r partially compensates for HIP1 function in the absence of HIP1 expression, providing strong evidence that HIP1 and HIP1r have overlapping roles in vivo. PMID- 15121853 TI - Groucho oligomerization is required for repression in vivo. AB - Drosophila Groucho (Gro) is a member of a family of metazoan corepressors with widespread roles in development. Previous studies indicated that a conserved domain in Gro, termed the Q domain, was required for repression in cultured cells and mediated homotetramerization. Evidence presented here suggests that the Q domain contains two coiled-coil motifs required for oligomerization and repression in vivo. Mutagenesis of the putative hydrophobic faces of these motifs, but not of the hydrophilic faces, prevents the formation of both tetramers and higher order oligomers. Mutagenesis of the hydrophobic faces of both coiled-coil motifs in the context of a Gal4-Gro fusion protein prevents repression of a Gal4-responsive reporter in S2 cells, while mutagenesis of a single motif weakens repression. The finding that the repression directed by the single mutants depends on endogenous wild-type Gro further supports the idea that oligomerization plays a role in repression. Overexpression in the fly of forms of Gro able to oligomerize, but not of a form of Gro unable to oligomerize, results in developmental defects and ectopic repression of Gro target genes in the wing disk. Although the function of several corepressors is suspected to involve oligomerization, these studies represent one of the first direct links between corepressor oligomerization and repression in vivo. PMID- 15121854 TI - Protein kinase C-alpha phosphorylation of specific serines in the connecting segment of the beta 4 integrin regulates the dynamics of type II hemidesmosomes. AB - Although the regulation of hemidesmosome dynamics during processes such as epithelial migration, wound healing, and carcinoma invasion is important, the mechanisms involved are poorly understood. The integrin alpha 6 beta 4 is an essential component of the hemidesmosome and a target of such regulation. Epidermal growth factor (EGF) can induce hemidesmosome disassembly by a mechanism that involves serine phosphorylation of the beta 4 integrin subunit. Using a combination of biochemical and mutational analyses, we demonstrate that EGF induces the phosphorylation of three specific serine residues (S(1356), S(1360), and S(1364)) located within the connecting segment of the beta 4 subunit and that phosphorylation on these residues accounts for the bulk of beta 4 phosphorylation stimulated by EGF. Importantly, phosphorylation of these serines is critical for the ability of EGF to disrupt hemidesmosomes. Using COS-7 cells, which assemble hemidesmosomes type II upon exogenous expression of the alpha 6 beta 4 integrin, we observed that expression of a beta 4 construct containing Ser-->Ala mutations of S(1356), S(1360), and S(1364) reduced the ability of EGF to disrupt hemidesmosomes and that this effect appears to involve cooperation among these phosphorylation sites. Moreover, expression of Ser-->Asp mutants that mimic constitutive phosphorylation reduced hemidesmosome formation. Protein kinase C alpha (PKC-alpha) is the kinase responsible for phosphorylating at least two of these serines, based on in vitro kinase assays, peptide mapping, and mutational analysis. Together, these results highlight the importance of serine phosphorylation in regulating type II hemidesmosome disassembly, implicate a cluster of serine residues within the connecting segment of beta 4, and argue for a key role for PKC-alpha in regulating these structures. PMID- 15121855 TI - Focal adhesion kinase suppresses apoptosis by binding to the death domain of receptor-interacting protein. AB - Tumor cells resist the apoptotic stimuli associated with invasion and metastasis by activating survival signals that suppress apoptosis. Focal adhesion kinase (FAK), a tyrosine kinase that is overexpressed in a variety of human tumors, mediates one of these survival signals. Attenuation of FAK expression in tumor cells results in apoptosis that is mediated by caspase 8- and FADD-dependent pathways, suggesting that death receptor pathways are involved in the process. Here, we report a functional link between FAK and death receptors. We have demonstrated that FAK binds to the death domain kinase receptor-interacting protein (RIP). RIP is a major component of the death receptor complex and has been shown to interact with Fas and tumor necrosis factor receptor 1 through its binding to adapter proteins. We have shown that RIP provides proapoptotic signals that are suppressed by its binding to FAK. We thus propose that FAK overexpression in human tumors provides a survival signal function by binding to RIP and inhibiting its interaction with the death receptor complex. PMID- 15121856 TI - Identification of PCIF1, a POZ domain protein that inhibits PDX-1 (MODY4) transcriptional activity. AB - Hox factors are evolutionarily conserved homeodomain-containing transcription factors that activate and repress gene expression in a precise temporally and spatially regulated manner during development and differentiation. Pancreatic duodenal homeobox 1 (PDX-1) is a Hox-type protein that is a critical requirement for normal pancreas development and for proper differentiation of the endocrine pancreas. In humans, PDX-1 gene mutation causes pancreatic agenesis and early- and late-onset type 2 diabetes. PDX-1 consists of an N-terminal transactivation domain, a homeodomain responsible for DNA binding and nuclear localization, and a conserved C terminus that is mutated in human diabetes but whose function is poorly understood. We have identified a novel POZ domain protein, PDX-1 C terminus-interacting factor 1 (PCIF1)/SPOP, that interacts with PDX-1 both in vitro and in vivo. PCIF1 is localized to the nucleus in a speckled pattern, and coexpression of PDX-1 alters the subnuclear distribution of PCIF1. Functionally, PCIF1 inhibits PDX-1 transactivation of established target gene promoters in a specific and dose-dependent manner that requires critical amino acids in the PDX 1 C terminus. PCIF1 is expressed in adult pancreatic insulin-producing beta cells, and overexpression of PCIF1 inhibits the rat insulin 1 and rat insulin 2 promoters in the MIN6 insulinoma beta cell line. The coexpression of PCIF1 with PDX-1 in beta cells and the ability of PCIF1 to repress PDX-1 transactivation suggest that modulation of PDX-1 function by PCIF1 may regulate normal beta cell differentiation. PMID- 15121857 TI - Sequential activation of phosphatidylinositol 3-kinase, beta Pix, Rac1, and Nox1 in growth factor-induced production of H2O2. AB - The generation of reactive oxygen species (ROS) in cells stimulated with growth factors requires the activation of phosphatidylinositol 3-kinase (PI3K) and the Rac protein. We report here that the COOH-terminal region of Nox1, a protein related to gp91(phox) (Nox2) of phagocytic cells, is constitutively associated with beta Pix, a guanine nucleotide exchange factor for Rac. Both growth factor induced ROS production and Rac1 activation were completely blocked in cells depleted of beta Pix by RNA interference. Rac1 was also shown to bind to the COOH terminal region of Nox1 in a growth factor-dependent manner. Moreover, the depletion of Nox1 by RNA interference inhibited growth factor-induced ROS generation. These results suggest that ROS production in growth factor-stimulated cells is mediated by the sequential activation of PI3K, beta Pix, and Rac1, which then binds to Nox1 to stimulate its NADPH oxidase activity. PMID- 15121858 TI - Nuclear export of phosphorylated galectin-3 regulates its antiapoptotic activity in response to chemotherapeutic drugs. AB - Galectin-3 (Gal-3), a member of the beta-galactoside binding protein family containing the NWGR antideath motif of the Bcl-2 protein family, is involved in various aspects of cancer progression. Previously, it has been shown that the antiapoptotic activity of Gal-3 is regulated by the phosphorylation at Ser(6) by casein kinase 1 (CK1). Here we questioned how phosphorylation at Ser(6) regulates Gal-3 function. We have generated serine-to-alanine (S6A) and serine-to-glutamic acid (S6E) Gal-3 mutants and transfected them into the BT-549 human breast carcinoma cell line, which does not express Gal-3. BT-549 cell clones expressing wild-type (wt) and mutant Gal-3 were exposed to chemotherapeutic anticancer drugs. In response to the apoptotic insults, phosphorylated wt Gal-3 was exported from the nucleus to the cytoplasm and protected the BT-549 cells from drug induced apoptosis while nonphosphorylated mutant Gal-3 neither was exported from the nucleus nor protected BT-549 cells from drug-induced apoptosis. Furthermore, leptomycin B, a nuclear export inhibitor, increased the cisplatin-induced apoptosis of Gal-3 expressing BT-549 cells. These results suggest that Ser(6) phosphoryaltion acts as a molecular switch for its cellular translocation from the nucleus to the cytoplasm and, as a result, regulates the antiapoptotic activity of Gal-3. PMID- 15121859 TI - The molecular scaffold KSR1 regulates the proliferative and oncogenic potential of cells. AB - The specificity of signaling through mitogen-activated protein kinase pathways has been attributed to both the control of intensity and duration of signaling and the actions of protein scaffolds. Here we demonstrate that the molecular scaffold KSR1 regulates the intensity and duration of ERK activation to modulate a cell's proliferative and oncogenic potential. Deletion of KSR1 eliminates the prolonged phase of ERK activation induced by platelet-derived growth factor and blocks Ras(V12)-induced transformation. The introduction of KSR1 into KSR1(-/-) mouse embryo fibroblasts causes a concentration-dependent increase in signaling and transformation, to a maximum at 14 times the wild-type KSR1 expression levels, but inhibits these responses at higher expression levels. An increase in KSR1 expression to levels that are optimal for signaling leads to a threefold increase in proliferative capacity and is coincident with the level of KSR1 expression that maximally associates with all members of the Raf/MEK/ERK cascade. These data reveal that cells contain a reserve proliferative capacity that is accessible by the optimal expression of a noncatalytic signaling component and that altering the expression level of a molecular scaffold can modulate the actions of growth factors and oncogenes. PMID- 15121861 TI - Reduced fertility in male mice deficient in the zinc metallopeptidase NL1. AB - Members of the M13 family of zinc metalloendopeptidases have been shown to play critical roles in the metabolism of various neuropeptides and peptide hormones, and they have been identified as important therapeutic targets. Recently, a mouse NL1 protein, a novel member of the family, was identified and shown to be expressed mainly in the testis as a secreted protein. To define its physiological role(s), we used a gene targeting strategy to disrupt the endogenous murine Nl1 gene by homologous recombination and generate Nl1 mutant mice. The Nl1(-/-) mice were viable and developed normally, suggesting that zygotic expression of Nl1 is not required for development. However, Nl1(-/-) males produced smaller litters than their wild-type siblings, indicating specific male fertility problems. Reduced fertility may be explained by two impaired processes, decreased egg fertilization and perturbed early development of fertilized eggs. These two phenotypes did not result from gross anatomical modifications of the testis or from impaired spermatogenesis. Basic sperm parameters were also normal. Thus, our findings suggest that one of the roles of NL1 in mice is related to sperm function and that NL1 modulates the processes of fertilization and early embryonic development in vivo. PMID- 15121860 TI - The transcription factor RFX3 directs nodal cilium development and left-right asymmetry specification. AB - There are five members of the RFX family of transcription factors in mammals. While RFX5 plays a well-defined role in the immune system, the functions of RFX1 to RFX4 remain largely unknown. We have generated mice with a deletion of the Rfx3 gene. RFX3-deficient mice exhibit frequent left-right (LR) asymmetry defects leading to a high rate of embryonic lethality and situs inversus in surviving adults. In vertebrates, specification of the LR body axis is controlled by monocilia in the embryonic node, and defects in nodal cilia consequently result in abnormal LR patterning. Consistent with this, Rfx3 is expressed in ciliated cells of the node and RFX3-deficient mice exhibit a pronounced defect in nodal cilia. In contrast to the case for wild-type embryos, for which we document for the first time a twofold increase in the length of nodal cilia during development, the cilia are present but remain markedly stunted in mutant embryos. Finally, we show that RFX3 regulates the expression of D2lic, the mouse orthologue of a Caenorhabditis elegans gene that is implicated in intraflagellar transport, a process required for the assembly and maintenance of cilia. In conclusion, RFX3 is essential for the differentiation of nodal monocilia and hence for LR body axis determination. PMID- 15121862 TI - Tumor necrosis factor alpha-induced apoptosis requires p73 and c-ABL activation downstream of RB degradation. AB - The retinoblastoma protein (RB) suppresses cell proliferation and apoptosis. We have previously shown that RB degradation is required for tumor necrosis factor alpha (TNF-alpha) to induce apoptosis. We show here the identification of two apoptotic effectors, i.e., c-ABL tyrosine kinase and p73, which are activated by TNF-alpha following RB degradation. In cells expressing a degradation-resistant RB protein (RB-MI), TNF-alpha does not activate c-ABL. RB-MI also inhibits TNF alpha-mediated activation of p73. Genetic deletion and pharmacological inhibition of c-ABL or p73 diminish the apoptotic response to TNF-alpha in human cell lines and mouse fibroblasts. Thymocytes isolated from Rb(MI/MI), Abl(-/-), or p73(-/-) mice are resistant to TNF-alpha-induced apoptosis compared to their wild-type counterparts. This is in contrast to p53(-/-) thymocytes, which exhibit a wild type level of apoptosis in response to TNF-alpha. Thus, c-ABL and p73 contribute to apoptosis induced by TNF-alpha, in addition to their role in promoting DNA damage-associated cell death. PMID- 15121863 TI - Dwarfism and impaired gut development in insulin-like growth factor II mRNA binding protein 1-deficient mice. AB - Insulin-like growth factor II mRNA-binding protein 1 (IMP1) belongs to a family of RNA-binding proteins implicated in mRNA localization, turnover, and translational control. Mouse IMP1 is expressed during early development, and an increase in expression occurs around embryonic day 12.5 (E12.5). To characterize the physiological role of IMP1, we generated IMP1-deficient mice carrying a gene trap insertion in the Imp1 gene. Imp1(-/-) mice were on average 40% smaller than wild-type and heterozygous sex-matched littermates. Growth retardation was apparent from E17.5 and remained permanent into adult life. Moreover, Imp1(-/-) mice exhibited high perinatal mortality, and only 50% were alive 3 days after birth. In contrast to most other organs, intestinal epithelial cells continue to express IMP1 postnatally, and Imp1(-/-) mice exhibited impaired development of the intestine, with small and misshapen villi and twisted colon crypts. Analysis of target mRNAs and global expression profiling at E12.5 indicated that Igf2 translation was downregulated, whereas the postnatal intestine showed reduced expression of transcripts encoding extracellular matrix components, such as galectin- 1, lumican, tenascin-C, procollagen transcripts, and the Hsp47 procollagen chaperone. Taken together, the results demonstrate that IMP1 is essential for normal growth and development. Moreover, IMP1 may facilitate intestinal morphogenesis via regulation of extracellular matrix formation. PMID- 15121864 TI - Reduced intranuclear mobility of APL fusion proteins accompanies their mislocalization and results in sequestration and decreased mobility of retinoid X receptor alpha. AB - Acute promyelocytic leukemia (APL) cells contain one of five chimeric retinoic acid alpha-receptor (RAR alpha) genes (X-RAR alpha) created by chromosomal translocations or deletion; each generates a fusion protein thought to transcriptionally repress RAR alpha target genes and block myeloid differentiation by an incompletely understood mechanism. To gain spatiotemporal insight into these oncogenic processes, we employed fluorescence microscopy and fluorescence recovery after photobleaching (FRAP). Fluorescence microscopy demonstrated that the intracellular localization of each of the X-RAR alpha proteins was distinct from that of RAR alpha and established which portion(s) of each X-RAR alpha protein-X, RAR, or both-contributed to its altered localization. Using FRAP, we demonstrated that the intranuclear mobility of each X-RAR alpha was reduced compared to that of RAR alpha. In addition, the mobility of each X RAR alpha was reduced further by ligand addition, in contrast to RAR alpha, which showed no change in mobility when ligand was added. Both the reduced baseline mobility of X-RAR alpha and the ligand-induced slowing of X-RAR alpha could be attributed to the protein interaction domain contained within X. RXR alpha aberrantly colocalized within each X-RAR alpha; colocalization of RXR alpha with promyelocytic leukemia (PML)-RAR alpha resulted in reduced mobility of RXR alpha. Thus, X-RAR alpha may interfere with RAR alpha through its aberrant nuclear dynamics, resulting in spatial and temporal sequestration of RXR alpha and perhaps other nuclear receptor coregulators critical for myeloid differentiation. PMID- 15121865 TI - The chromatin-remodeling BAF complex mediates cellular antiviral activities by promoter priming. AB - The elicitation of cellular antiviral activities is dependent on the rapid transcriptional activation of interferon (IFN) target genes. It is not clear how the interferon target promoters, which are organized into chromatin structures in cells, rapidly respond to interferon or viral stimulation. In this report, we show that alpha IFN (IFN-alpha) treatment of HeLa cells induced hundreds of genes. The induction of the majority of these genes was inhibited when one critical subunit of the chromatin-remodeling SWI/SNF-like BAF complexes, BAF47, was knocked down via RNA interference. Inhibition of BAF47 blocked the cellular response to viral infection and impaired cellular antiviral activity by inhibiting many IFN- and virus-inducible genes. We show that the BAF complex was required to mediate both the basal-level expression and the rapid induction of the antiviral genes. Further analyses indicated that the BAF complex primed some IFN target promoters by utilizing ATP-derived energy to maintain the chromatin in a constitutively open conformation, allowing faster and more potent induction after IFN-alpha treatment. We propose that constitutive binding of the BAF complex is an important mechanism for the IFN-inducible promoters to respond rapidly to IFN and virus stimulation. PMID- 15121866 TI - Wnt-3a and Dvl induce neurite retraction by activating Rho-associated kinase. AB - Dvl is a key protein that transmits the Wnt signal to the canonical beta-catenin pathway and the noncanonical planar cell polarity (PCP) pathway. We studied the roles of Rho-associated kinase (Rho-kinase), which is activated by Dvl in the PCP pathway of mammalian cells. The expression of Dvl-1, Wnt-1, or Wnt-3a activated Rho-kinase in COS cells, and this activation was inhibited by the Rho-binding domain of Rho-kinase. The expression of Dvl-1 in PC12 cells activated Rho and inhibited nerve growth factor (NGF)-induced neurite outgrowth. This inhibition was reversed by a Rho-kinase inhibitor but not by a c-Jun N-terminal kinase inhibitor. Dvl-1 also inhibited serum starvation-dependent neurite outgrowth of N1E-115 cells, and expression of the Rho-binding domain of Rho-kinase reversed this inhibitory activity of Dvl-1. Dvl-1 mutants that did not activate Rho-kinase did not inhibit the neurite outgrowth of N1E-115 cells. Furthermore, the purified Wnt-3a protein activated Rho-kinase and inhibited the NGF-dependent neurite outgrowth of PC12 cells. Wnt-3a-dependent neurite retraction was also prevented by a Rho-kinase inhibitor and a Dvl-1 mutant that suppresses Wnt-3a-dependent activation of Rho-kinase. These results suggest that Wnt-3a and Dvl regulate neurite formation through Rho-kinase and that PC12 and N1E-115 cells are useful for analyzing the PCP pathway. PMID- 15121867 TI - TRAF family proteins link PKR with NF-kappa B activation. AB - The double-stranded RNA (dsRNA)-dependent protein kinase PKR activates NF-kappa B via the I kappa B kinase (IKK) complex, but little is known about additional molecules that may be involved in this pathway. Analysis of the PKR sequence enabled us to identify two putative TRAF-interacting motifs. The viability of such an interaction was further suggested by computer modeling. Here, we present evidence of the colocalization and physical interaction between PKR and TRAF family proteins in vivo, as shown by immunoprecipitation and confocal microscopy experiments. This interaction is induced upon PKR dimerization. Most importantly, we show that the binding between PKR and TRAFs is functionally relevant, as observed by the absence of NF-kappa B activity upon PKR expression in cells genetically deficient in TRAF2 and TRAF5 or after expression of TRAF dominant negative molecules. On the basis of sequence information and mutational and computer docking analyses, we favored a TRAF-PKR interaction model in which the C terminal domain of TRAF binds to a predicted TRAF interaction motif present in the PKR kinase domain. Altogether, our data suggest that TRAF family proteins are key components located downstream of PKR that have an important role in mediating activation of NF-kappa B by the dsRNA-dependent protein kinase. PMID- 15121868 TI - Specification of the retinal fate of mouse embryonic stem cells by ectopic expression of Rx/rax, a homeobox gene. AB - With the goal of generating retinal cells from mouse embryonic stem (ES) cells by exogenous gene transfer, we introduced the Rx/rax transcription factor, which is expressed in immature retinal cells, into feeder-free mouse ES cells (CCE). CCE cells expressing Rx/rax as well as enhanced green fluorescent protein (CCE-RX/E cells) proliferated and remained in the undifferentiated state in the presence of leukemia inhibitory factor, as did parental ES cells. We made use of mouse embryo retinal explant cultures to address the differentiation ability of grafted ES cells. Dissociated embryoid bodies were treated with retinoic acid for use as donor cells and cocultured with retina explants for 2 weeks. In contrast to the parental CCE cells, which could not migrate into host retinal cultures, CCE-RX/E cells migrated into the host retina and extended their process-like structures between the host retinal cells. Most of the grafted CCE-RX/E cells became located in the ganglion cell and inner plexiform layers and expressed ganglion and horizontal cell markers. Furthermore, these grafted cells had the electrophysiological properties expected of ganglion cells. Our data thus suggest that subpopulations of retinal neurons can be generated in retinal explant cultures from grafted mouse ES cells ectopically expressing the transcription factor Rx/rax. PMID- 15121869 TI - The Herpesvirus saimiri small nuclear RNAs recruit AU-rich element-binding proteins but do not alter host AU-rich element-containing mRNA levels in virally transformed T cells. AB - Herpesvirus saimiri (HVS) encodes seven Sm-class small nuclear RNAs, called HSURs (for Herpesvirus saimiri U RNAs), that are abundantly expressed in HVS transformed, latently infected marmoset T cells but are of unknown function. HSURs 1, 2, and 5 have highly conserved 5'-end sequences containing the AUUUA pentamer characteristic of AU-rich elements (AREs) that regulate the stability of many host mRNAs, including those encoding most proto-oncogenes and cytokines. To test whether the ARE-containing HSURs act to sequester host proteins that regulate the decay of these mRNAs, we demonstrate their in vivo interaction with the ARE-binding proteins hnRNP D and HuR in HVS-transformed T cells using a new cross-linking assay. Comprehensive Northern and microarray analyses revealed, however, that the levels of endogenous ARE-containing mRNAs are not altered in T cells latently infected with HVS mutants lacking HSURs 1 and 2. HSUR 1 binds the destabilizing ARE-binding protein tristetraprolin induced following activation of HVS-transformed T cells, but even in such stimulated cells, the levels of host ARE-containing mRNAs are not altered by deletion of HSURs 1 and 2. Instead, HSUR 1 itself is degraded by an ARE-dependent pathway in HVS-transformed T cells, suggesting that HVS may take advantage of the host ARE-mediated mRNA decay pathway to regulate HSUR expression. This is the first example of posttranscriptional regulation of the expression of an Sm small nuclear RNA. PMID- 15121870 TI - Extracellular signal-regulated kinase induces the megakaryocyte GPIIb/CD41 gene through MafB/Kreisler. AB - Extracellular signal-regulated kinase (ERK) facilitates cell cycle progression in most mammalian cells, but in certain cell types prolonged signaling through this pathway promotes differentiation and lineage-specific gene expression through mechanisms that are poorly understood. Here, we characterize the transcriptional regulation of platelet GPIIb integrin (CD41) by ERK during megakaryocyte differentiation. ERK-dependent transactivation involves the proximal promoter of GPIIb within 114 bp upstream of the transcriptional start site. GATA, Ets, and Sp1 consensus sequences within this region are each necessary and function combinatorially in ERK-activated transcription. MafB/Kreisler is induced in response to ERK and synergizes with GATA and Ets to enhance transcription from the proximal promoter. The requirement for MafB in promoter regulation is demonstrated by inhibition of transactivation following dominant-negative or antisense suppression of MafB function. Thus, ERK promotes megakaryocyte differentiation by coordinate regulation of nuclear factors that synergize in GPIIb promoter regulation. These results establish a novel role for MafB as a regulator of ERK-induced gene expression. PMID- 15121871 TI - E2F-dependent histone acetylation and recruitment of the Tip60 acetyltransferase complex to chromatin in late G1. AB - E2F proteins can either activate or repress transcription. Following mitogenic stimulation, repressive E2F4-p130-histone deacetylase complexes dissociate from, while activating species (E2F1, -2, and -3) associate with, target promoters. Histones H3 and H4 simultaneously become hyperacetylated, but it remains unclear whether this is a prerequisite or a consequence of E2F binding. Here, we show that activating E2F species are required for hyperacetylation of target chromatin in human cells. Overexpression of a dominant-negative (DN) E2F1 mutant in serum stimulated T98G cells blocked all E2F binding, H4 acetylation, and, albeit partially, H3 acetylation. Target gene activation and S-phase entry were also blocked by DN E2F1. Conversely, ectopic activation of E2F1 rapidly induced H3 and H4 acetylation, demonstrating a direct role for E2F in these events. E2F1 was previously shown to bind the histone acetyltransferases (HATs) p300/CBP and PCAF/GCN5. In our hands, ectopically expressed E2F1 also bound the unrelated HAT Tip60 and induced recruitment of five subunits of the Tip60 complex (Tip60, TRRAP, p400, Tip48, and Tip49) to target promoters in vivo. Moreover, E2F dependent recruitment of Tip60 to chromatin occurred in late G(1) following serum stimulation. We speculate that the activities of multiple HAT complexes account for E2F-dependent acetylation, transcription, and S-phase entry. PMID- 15121872 TI - Tyrosine 813 is a site of JAK2 autophosphorylation critical for activation of JAK2 by SH2-B beta. AB - The tyrosine kinase Janus kinase 2 (JAK2) binds to the majority of the known members of the cytokine family of receptors. Ligand-receptor binding leads to activation of the associated JAK2 molecules, resulting in rapid autophosphorylation of multiple tyrosines within JAK2. Phosphotyrosines can then serve as docking sites for downstream JAK2 signaling molecules. Despite the importance of these phosphotyrosines in JAK2 function, only a few sites and binding partners have been identified. Using two-dimensional phosphopeptide mapping and a phosphospecific antibody, we identified tyrosine 813 as a site of JAK2 autophosphorylation of overexpressed JAK2 and endogenous JAK2 activated by growth hormone. Tyrosine 813 is contained within a YXXL sequence motif associated with several other identified JAK2 phosphorylation sites. We show that phosphorylation of tyrosine 813 is required for the SH2 domain-containing adapter protein SH2-B beta to bind JAK2 and to enhance the activity of JAK2 and STAT5B. The homologous tyrosine in JAK3, tyrosine 785, is autophosphorylated in response to interleukin-2 stimulation and is required for SH2-B beta to bind JAK3. Taken together these data strongly suggest that tyrosine 813 is a site of autophosphorylation in JAK2 and is the SH2-B beta-binding site within JAK2 that is required for SH2-B beta to enhance activation of JAK2. PMID- 15121873 TI - Modification of subtelomeric DNA. AB - There is a discrepancy in telomere length as measured by signal intensity of telomere restriction fragments on gels and fluorescence in situ hybridization analysis. This difference has been ascribed to the X-region, a segment of subtelomeric DNA that is resistant to being cut by restriction enzymes. To explore the nature of this region, we analyzed the digestibility of an artificial seeded telomere in HeLa cells as well as the Xp/Yp autosomal telomere in human BJ fibroblasts. We found that there is a substantial fraction of subtelomeric DNA containing restriction sites that is not digested with enzymes such as EcoRI, NlaIII, and SphI. Comparison of methylation-sensitive and -resistant enzymes excluded the possibility of the X-region being maintained by DNA methylation. We show that the X-region represents a variable domain whose size changes with telomere length, and neither non-TTAGGG sequences nor cytidine methylation can adequately explain the size of the X-region. PMID- 15121874 TI - The adapter molecule Sin regulates T-cell-receptor-mediated signal transduction by modulating signaling substrate availability. AB - Engagement of the T-cell receptor (TCR) results in the activation of a multitude of signaling events that regulate the function of T lymphocytes. These signaling events are in turn modulated by adapter molecules, which control the final functional output through the formation of multiprotein complexes. In this report, we identified the adapter molecule Sin as a new regulator of T-cell activation. We found that the expression of Sin in transgenic T lymphocytes and Jurkat T cells inhibited interleukin-2 expression and T-cell proliferation. This inhibitory effect was specific and was due to defective phospholipase C-gamma (PLC-gamma) phosphorylation and activation. In contrast to other adapters that become phosphorylated upon TCR stimulation, Sin was constitutively phosphorylated in resting cells by the Src kinase Fyn and bound to signaling intermediates, including PLC-gamma. In stimulated cells, Sin was transiently dephosphorylated, which coincided with transient dissociation of Fyn and PLC-gamma. Downregulation of Sin expression using Sin-specific short interfering RNA oligonucleotides inhibited transcriptional activation in response to TCR stimulation. Our results suggest that endogenous Sin influences T-lymphocyte signaling by sequestering signaling substrates and regulating their availability and/or activity in resting cells, while Sin is required for targeting these intermediates to the TCR for fast signal transmission during stimulation. PMID- 15121875 TI - Acquisition of Hrs, an essential component of phagosomal maturation, is impaired by mycobacteria. AB - Pathogenic mycobacteria survive within macrophages by precluding the fusion of phagosomes with late endosomes or lysosomes. Because the molecular determinants of normal phagolysosome formation are poorly understood, the sites targeted by mycobacteria remain unidentified. We found that Hrs, an adaptor molecule involved in protein sorting, associates with phagosomes prior to their fusion with late endosomes or lysosomes. Recruitment of Hrs required the interaction of its FYVE domain with phagosomal phosphatidylinositol 3-phosphate, but two other attachment sites were additionally involved. Depletion of Hrs by use of small interfering RNA impaired phagosomal maturation, preventing the acquisition of lysobisphosphatidic acid and reducing luminal acidification. As a result, the maturation of phagosomes formed in Hrs-depleted cells was arrested at an early stage, characterized by the acquisition and retention of sorting endosomal markers. This phenotype is strikingly similar to that reported to occur in phagosomes of cells infected by mycobacteria. We therefore tested whether Hrs is recruited to phagosomes containing mycobacteria. Hrs associated readily with phagosomes containing inert particles but poorly with mycobacterial phagosomes. Moreover, Hrs was found more frequently in phagosomes containing avirulent Mycobacterium smegmatis than in phagosomes with the more virulent Mycobacterium marinum. These findings suggest that the inability to recruit Hrs contributes to the arrest of phagosomal maturation induced by pathogenic mycobacteria. PMID- 15121876 TI - Effect of mifepristone on the expression of cytokines in the human Fallopian tube. AB - Cytokines are believed to play a critical role as mediators between the oviduct and the developing embryo. A synchronous development of embryo and endometrium is essential to successful implantation. It seems to be beneficial for embryo development to rest for some time in the Fallopian tube. Expression of cytokines in the human Fallopian tube and the effect of mifepristone were investigated. Fourteen women with regular menstrual cycles and proven fertility, admitted to the hospital for tubal ligation, were randomly allocated to control or treatment groups. Mifepristone 200 mg was given on day LH+2. Surgery was performed on day LH+3 to LH+5. Biopsies were obtained from the ampullar and isthmic regions of the tubes. Expression of interleukin 8 (IL-8), tumour necrosis factor alpha (TNFalpha), transforming growth factor beta (TGFbeta) and leukaemia inhibitory factor (LIF) was analysed using immunohistochemistry. All cytokines except IL-8 showed the same staining intensity both in the ampullar and isthmic region, while IL-8 was more pronounced in the ampullar region in both epithelial and stromal cells. Exposure to mifepristone made the spatial difference in IL-8 disappear and increased the expression of TNFalpha in the epithelium of the isthmus, but had no effect on the expression of TGFbeta1 or LIF. Changes in cytokine expression in the Fallopian tube are likely to influence embryo development, which could contribute to the contraceptive effect of mifepristone. PMID- 15121877 TI - Comparative analysis between cyclic GMP and cyclic AMP signalling in human sperm. AB - The principal involvement of cyclic nucleotides in regulating sperm functions is well established, but the factors controlling their generation and actions have not yet been entirely resolved. In particular, specific roles for cyclic (c)GMP in mammalian sperm are poorly understood. In this study, we have characterized comparatively the cAMP and cGMP signalling systems in ejaculated human sperm. Mean concentrations of cGMP (0.1 micromol/l) were found to be 100-fold lower than those of cAMP in non-stimulated cells, and adenylyl cyclase (AC) activities predominate by far guanylyl cyclase (GC) activities in both particulate and soluble protein fractions. By different experimental approaches (photoaffinity labelling, cyclase assays, immunoblotting), we provide evidence for the presence (guanylyl cyclase-A, soluble guanylyl cyclase, regulatory and catalytic subunits of cAMP-dependent protein kinase) or absence (guanylyl cyclase-B, natriuretic peptide clearance receptor, neuronal nitric oxide synthase, cGMP-dependent protein kinase I) of different factors involved in either cAMP or cGMP pathways. Functional studies showed that cGMP, at high concentrations, can enhance sperm protein tyrosine phosphorylation but not serine phosphorylation of glycogen synthase kinase. This study reveals that human sperm are characterized by an exceptional predominance of cAMP signalling and indicates potential roles for cGMP. PMID- 15121878 TI - Estradiol abrogates apoptosis in breast cancer cells through inactivation of BAD: Ras-dependent nongenomic pathways requiring signaling through ERK and Akt. AB - Estrogens such as 17-beta estradiol (E(2)) play a critical role in sporadic breast cancer progression and decrease apoptosis in breast cancer cells. Our studies using estrogen receptor-positive MCF7 cells show that E(2) abrogates apoptosis possibly through phosphorylation/inactivation of the proapoptotic protein BAD, which was rapidly phosphorylated at S112 and S136. Inhibition of BAD protein expression with specific antisense oligonucleotides reduced the effectiveness of tumor necrosis factor-alpha, H(2)O(2), and serum starvation in causing apoptosis. Furthermore, the ability of E(2) to prevent tumor necrosis factor-alpha-induced apoptosis was blocked by overexpression of the BAD S112A/S136A mutant but not the wild-type BAD. BAD S112A/S136A, which lacks phosphorylation sites for p90(RSK1) and Akt, was not phosphorylated in response to E(2) in vitro(.) E(2) treatment rapidly activated phosphatidylinositol 3 kinase (PI-3K)/Akt and p90(RSK1) to an extent similar to insulin-like growth factor-1 treatment. In agreement with p90(RSK1) activation, E(2) also rapidly activated extracellular signal-regulated kinase, and this activity was down regulated by chemical and biological inhibition of PI-3K suggestive of cross talk between signaling pathways responding to E(2). Dominant negative Ras blocked E(2) induced BAD phosphorylation and the Raf-activator RasV12T35S induced BAD phosphorylation as well as enhanced E(2)-induced phosphorylation at S112. Chemical inhibition of PI-3K and mitogen-activated protein kinase kinase 1 inhibited E(2)-induced BAD phosphorylation at S112 and S136 and expression of dominant negative Ras-induced apoptosis in proliferating cells. Together, these data demonstrate a new nongenomic mechanism by which E(2) prevents apoptosis. PMID- 15121879 TI - Multiple interactions of rad23 suggest a mechanism for ubiquitylated substrate delivery important in proteolysis. AB - The mechanism underlying the delivery of ubiquitylated substrates to the proteasome is poorly understood. Rad23 is a putative adaptor molecule for this process because it interacts with ubiquitin chains through its ubiquitin associated motifs (UBA) and with the proteasome through a ubiquitin-like element (UBL). Here, we demonstrate that the UBL motif of Rad23 also binds Ufd2, an E4 enzyme essential for ubiquitin chain assembly onto its substrates. Mutations in the UBL of Rad23 alter its interactions with Ufd2 and the proteasome, and impair its function in the UFD proteolytic pathway. Furthermore, Ufd2 and the proteasome subunit Rpn1 compete for the binding of Rad23, suggesting that Rad23 forms separate complexes with them. Importantly, we also find that the ability of other UBL/UBA proteins to associate with Ufd2 correlates with their differential involvement in the UFD pathway, suggesting that UBL-mediated interactions may contribute to the substrate specificity of these adaptors. We propose that the UBL motif, a protein-protein interaction module, may be used to facilitate coupling between substrate ubiquitylation and delivery, and to ensure the orderly handoff of the substrate from the ubiquitylation machinery to the proteasome. PMID- 15121880 TI - Salmonella impairs RILP recruitment to Rab7 during maturation of invasion vacuoles. AB - After invasion of epithelial cells, Salmonella enterica Typhimurium resides within membrane-bound vacuoles where it survives and replicates. Like endocytic vesicles, the Salmonella-containing vacuoles (SCVs) undergo a maturation process that involves sequential acquisition of Rab5 and Rab7 and displacement toward the microtubule-organizing center. However, SCVs fail to merge with lysosomes and instead develop subsequently into a filamentous network that extends toward the cell periphery. We found that the initial centripetal displacement of the SCV is due to recruitment by Rab7 of Rab7-interacting lysosomal protein (RILP), an effector protein that can simultaneously associate with the dynein motor complex. Unlike the early SCVs, the Salmonella-induced filaments (Sifs) formed later are devoid of RILP and dynein, despite the presence of active Rab7 on their membranes. Kinesin seems to be involved in the elongation of Sifs. SifA, a secreted effector of Salmonella, was found to be at least partly responsible for uncoupling Rab7 from RILP in Sifs and in vitro experiments suggest that SifA may exert this effect by interacting with Rab7. We propose that, by disengaging RILP from Rab7, SifA enables the centrifugal extension of tubules from the Salmonella containing vacuoles, thereby providing additional protected space for bacterial replication. PMID- 15121881 TI - Disturbed cholesterol traffic but normal proteolytic function in LAMP-1/LAMP-2 double-deficient fibroblasts. AB - Mice double deficient in LAMP-1 and -2 were generated. The embryos died between embryonic days 14.5 and 16.5. An accumulation of autophagic vacuoles was detected in many tissues including endothelial cells and Schwann cells. Fibroblast cell lines derived from the double-deficient embryos accumulated autophagic vacuoles and the autophagy protein LC3II after amino acid starvation. Lysosomal vesicles were larger and more peripherally distributed and showed a lower specific density in Percoll gradients in double deficient when compared with control cells. Lysosomal enzyme activities, cathepsin D processing and mannose-6-phosphate receptor expression levels were not affected by the deficiency of both LAMPs. Surprisingly, LAMP-1 and -2 deficiencies did not affect long-lived protein degradation rates, including proteolysis due to chaperone-mediated autophagy. The LAMP-1/2 double-deficient cells and, to a lesser extent, LAMP-2 single-deficient cells showed an accumulation of unesterified cholesterol in endo/lysosomal, rab7, and NPC1 positive compartments as well as reduced amounts of lipid droplets. The cholesterol accumulation in LAMP-1/2 double-deficient cells could be rescued by overexpression of murine LAMP-2a, but not by LAMP-1, highlighting the more prominent role of LAMP-2. Taken together these findings indicate partially overlapping functions for LAMP-1 and -2 in lysosome biogenesis, autophagy, and cholesterol homeostasis. PMID- 15121882 TI - Sorting nexin 17 accelerates internalization yet retards degradation of P selectin. AB - The transient appearance of P-selectin on the surface of endothelial cells helps recruit leukocytes into sites of inflammation. The tight control of cell surface P-selectin on these cells depends on regulated exocytosis of Weibel-Palade bodies where the protein is stored and on its rapid endocytosis. After endocytosis, P selectin is either sorted via endosomes and the Golgi apparatus for storage in Weibel-Palade bodies or targeted to lysosomes for degradation. A potential player in this complex endocytic itinerary is SNX17, a member of the sorting nexin family, which has been shown in a yeast two-hybrid assay to bind P-selectin. Here, we show that overexpression of SNX17 in mammalian cells can influence two key steps in the endocytic trafficking of P-selectin. First, it promotes the endocytosis of P-selectin from the plasma membrane. Second, it inhibits the movement of P-selectin into lysosomes, thereby reducing its degradation. PMID- 15121883 TI - A human telomerase-associated nuclease. AB - Ciliate and yeast telomerase possess a nucleolytic activity capable of removing DNA from the 3' end of a single-stranded oligonucleotide substrate. The nuclease activity is thought to assist in enzyme proofreading and/or processivity. Herein, we report a previously uncharacterized human telomerase-associated nuclease activity that shares several properties with ciliate and yeast telomerases. Partially purified human telomerase, either from cell extracts or recombinantly produced, demonstrated an ability to remove 3' nontelomeric nucleotides from a substrate containing 5' telomeric DNA, followed by extension of the newly exposed telomeric sequence. This cleavage/extension activity was apparent at more than one position within the telomeric DNA and was influenced by sequences 5' to the telomeric/nontelomeric boundary and by substitution with a methylphosphonate moiety at the telomeric/nontelomeric DNA boundary. Our data suggest that human telomerase is associated with an evolutionarily conserved nucleolytic activity and support a model in which telomerase-substrate interactions can occur distal from the 3' primer end. PMID- 15121884 TI - Intracellular trafficking of bile salt export pump (ABCB11) in polarized hepatic cells: constitutive cycling between the canalicular membrane and rab11-positive endosomes. AB - The bile salt export pump (BSEP, ABCB11) couples ATP hydrolysis with transport of bile acids into the bile canaliculus of hepatocytes. Its localization in the apical canalicular membrane is physiologically regulated by the demand to secrete biliary components. To gain insight into how such localization is regulated, we studied the intracellular trafficking of BSEP tagged with yellow fluorescent protein (YFP) in polarized WIF-B9 cells. Confocal imaging revealed that BSEP-YFP was localized at the canalicular membrane and in tubulo-vesicular structures either adjacent to the microtubule-organizing center or widely distributed in the cytoplasm. In the latter two locations, BSEP-YFP colocalized with rab11, an endosomal marker. Selective photobleaching experiments revealed that single BSEP YFP molecules resided in canalicular membranes only transiently before exchanging with intracellular BSEP-YFP pools. Such exchange was inhibited by microtubule and actin inhibitors and was unaffected by brefeldin A, dibutyryl cyclic AMP, taurocholate, or PI 3-kinase inhibitors. Intracellular carriers enriched in BSEP YFP elongated and dissociated as tubular elements from a globular structure adjacent to the microtubule-organizing center. They displayed oscillatory movement toward either canalicular or basolateral membranes, but only fused with the canalicular membrane. The pathway between canalicular and intracellular membranes that BSEP constitutively cycles within could serve to regulate apical pools of BSEP as well as other apical membrane transporters. PMID- 15121885 TI - A ubiquitous beta-tubulin disrupts microtubule assembly and inhibits cell proliferation. AB - Vertebrate tubulin is encoded by a multigene family that produces distinct gene products, or isotypes, of both the alpha- and beta-tubulin subunits. The isotype sequences are conserved across species supporting the hypothesis that different isotypes subserve different functions. To date, however, most studies have demonstrated that tubulin isotypes are freely interchangeable and coassemble into all classes of microtubules. We now report that, in contrast to other isotypes, overexpression of a mouse class V beta-tubulin cDNA in mammalian cells produces a strong, dose-dependent disruption of microtubule organization, increased microtubule fragmentation, and a concomitant reduction in cellular microtubule polymer levels. These changes also disrupt mitotic spindle assembly and block cell proliferation. Consistent with diminished microtubule assembly, there is an increased tolerance for the microtubule stabilizing drug, paclitaxel, which is able to reverse many of the effects of class V beta-tubulin overexpression. Moreover, transfected cells selected in paclitaxel exhibit increased expression of class V beta-tubulin, indicating that this isotype is responsible for the drug resistance. The results show that class V beta-tubulin is functionally distinct from other tubulin isotypes and imparts unique properties on the microtubules into which it incorporates. PMID- 15121895 TI - Three-dimensional motifs from the SCOR, structural classification of RNA database: extruded strands, base triples, tetraloops and U-turns. AB - Release 2.0.1 of the Structural Classification of RNA (SCOR) database, http://scor.lbl.gov, contains a classification of the internal and hairpin loops in a comprehensive collection of 497 NMR and X-ray RNA structures. This report discusses findings of the classification that have not been reported previously. The SCOR database contains multiple examples of a newly described RNA motif, the extruded helical single strand. Internal loop base triples are classified in SCOR according to their three-dimensional context. These internal loop triples contain several examples of a frequently found motif, the minor groove AGC triple. SCOR also presents the predominant and alternate conformations of hairpin loops, as shown in the most well represented tetraloops, with consensus sequences GNRA, UNCG and ANYA. The ubiquity of the GNRA hairpin turn motif is illustrated by its presence in complex internal loops. PMID- 15121896 TI - Identification and functional analysis of 'hypothetical' genes expressed in Haemophilus influenzae. AB - The progress in genome sequencing has led to a rapid accumulation in GenBank submissions of uncharacterized 'hypothetical' genes. These genes, which have not been experimentally characterized and whose functions cannot be deduced from simple sequence comparisons alone, now comprise a significant fraction of the public databases. Expression analyses of Haemophilus influenzae cells using a combination of transcriptomic and proteomic approaches resulted in confident identification of 54 'hypothetical' genes that were expressed in cells under normal growth conditions. In an attempt to understand the functions of these proteins, we used a variety of publicly available analysis tools. Close homologs in other species were detected for each of the 54 'hypothetical' genes. For 16 of them, exact functional assignments could be found in one or more public databases. Additionally, we were able to suggest general functional characterization for 27 more genes (comprising approximately 80% total). Findings from this analysis include the identification of a pyruvate-formate lyase-like operon, likely to be expressed not only in H.influenzae but also in several other bacteria. Further, we also observed three genes that are likely to participate in the transport and/or metabolism of sialic acid, an important component of the H.influenzae lipo-oligosaccharide. Accurate functional annotation of uncharacterized genes calls for an integrative approach, combining expression studies with extensive computational analysis and curation, followed by eventual experimental verification of the computational predictions. PMID- 15121897 TI - DNA binding and antigene activity of a daunomycin-conjugated triplex-forming oligonucleotide targeting the P2 promoter of the human c-myc gene. AB - Triplex-forming oligonucleotides (TFO) that bind DNA in a sequence-specific manner might be used as selective repressors of gene expression and gene-targeted therapeutics. However, many factors, including instability of triple helical complexes in cells, limit the efficacy of this approach. In the present study, we tested whether covalent linkage of a TFO to daunomycin, which is a potent DNA intercalating agent and anticancer drug, could increase stability of the triple helix and activity of the oligonucleotide in cells. The 11mer daunomycin conjugated GT (dauno-GT11) TFO targeted a sequence upstream of the P2 promoter, a site known to be critical for transcription of the c-myc gene. Band-shift assays showed that the dauno-GT11 formed triplex DNA with enhanced stability compared to the unmodified TFO. Band shift and footprinting experiments demonstrated that binding of dauno-GT11 was highly sequence-specific with exclusive binding to the 11 bp target site in the c-myc promoter. The daunomycin-conjugated TFO inhibited transcription in vitro and reduced c-myc promoter activity in prostate and breast cancer cells. The daunomycin-conjugated TFO was taken up by cells with a distinctive intracellular distribution compared to free daunomycin. However, cationic lipid-mediated delivery was required for enhanced cellular uptake, nuclear localization and biological activity of the TFO in cells. Dauno-GT11 reduced transcription of the endogenous c-myc gene in cells, but did not affect expression of non-target genes, such as ets-1 and ets-2, which contained very similar target sequences in their promoters. Daunomycin-conjugated control oligonucleotides unable to form triplex DNA with the target sequence did not have any effect in these assays, indicating that daunomycin was not directly responsible for the activity of daunomycin-conjugated TFO. Thus, attachment of daunomycin resulted in increased triplex stability and biological activity of the 11mer GT-rich TFO without compromising its specificity. These results encourage further testing of this approach to develop novel antigene therapeutics. PMID- 15121898 TI - The composition of Staufen-containing RNA granules from human cells indicates their role in the regulated transport and translation of messenger RNAs. AB - hStaufen is the human homolog of dmStaufen, a double-stranded (ds)RNA-binding protein involved in early development of the fly. hStaufen-containing complexes were purified by affinity chromatography from human cells transfected with a TAP tagged hStaufen gene. These complexes showed a size >10 MDa. Untagged complexes with similar size were identified from differentiated human neuroblasts. The identity of proteins present in purified hStaufen complexes was determined by mass spectrometry and the presence of these proteins and other functionally related ones was verified by western blot. Ribosomes and proteins involved in the control of protein synthesis (PABP1 and FMRP) were present in purified hStaufen complexes, as well as elements of the cytoskeleton (tubulins, tau, actin and internexin), cytoskeleton control proteins (IQGAP1, cdc42 and rac1) and motor proteins (dynein, kinesin and myosin). In addition, proteins normally found in the nucleus, like nucleolin and RNA helicase A, were also found associated with cytosolic hStaufen complexes. The co-localization of these components with hStaufen granules in the dendrites of differentiated neuroblasts, determined by confocal immunofluorescence, validated their association in living cells. These results support the notion that the hStaufen-containing granules are structures essential in the localization and regulated translation of human mRNAs in vivo. PMID- 15121899 TI - APOBEC3G is a single-stranded DNA cytidine deaminase and functions independently of HIV reverse transcriptase. AB - In the absence of the viral vif gene, human immunodeficiency virus (HIV) may be restricted by the APOBEC3G gene on chromosome 22. The role of the HIV Vif protein is to exclude host cell APOBEC3G from the budding virion. As APOBEC3G shows sequence homology to cytidine deaminases, it is presumed that in the absence of Vif, cytidine residues in the cDNA are deaminated yielding uracil. It is not known if additional proteins mediate APOBEC3G function or if deamination occurs in concert with reverse transcription. This report describes an in vitro assay showing that Baculovirus derived APOBEC3G alone extensively deaminates cDNA independently of reverse transcriptase. It reproduces the dinucleotide context typical of G --> A hypermutants derived from a Delta(vif) virus. By using an RNaseH- form of reverse transcriptase, it was shown that the cDNA has to be free of its RNA template to allow deamination. APOBEC3G deamination of dC or dCTP was not detected. In short, APOBEC3G is a single-stranded DNA cytidine deaminase capable of restricting retroviral replication. PMID- 15121900 TI - Characterization of the 3' exonuclease subunit DP1 of Methanococcus jannaschii replicative DNA polymerase D. AB - The B-subunits associated with the replicative DNA polymerases are conserved from Archaea to humans, whereas the corresponding catalytic subunits are not related. The latter belong to the B and D DNA polymerase families in eukaryotes and archaea, respectively. Sequence analysis places the B-subunits within the calcineurin-like phosphoesterase superfamily. Since residues implicated in metal binding and catalysis are well conserved in archaeal family D DNA polymerases, it has been hypothesized that the B-subunit could be responsible for the 3'-5' proofreading exonuclease activity of these enzymes. To test this hypothesis we expressed Methanococcus jannaschii DP1 (MjaDP1), the B-subunit of DNA polymerase D, in Escherichia coli, and demonstrate that MjaDP1 functions alone as a moderately active, thermostable, Mn2+-dependent 3'-5' exonuclease. The putative polymerase subunit DP2 is not required. The nuclease activity is strongly reduced by single amino acid mutations in the phosphoesterase domain indicating the requirement of this domain for the activity. MjaDP1 acts as a unidirectional, non processive exonuclease preferring mispaired nucleotides and single-stranded DNA, suggesting that MjaDP1 functions as the proofreading exonuclease of archaeal family D DNA polymerase. PMID- 15121901 TI - Multiple protein/protein and protein/RNA interactions suggest roles for yeast DNA/RNA helicase Sen1p in transcription, transcription-coupled DNA repair and RNA processing. AB - Sen1p in Saccharomyces cerevisiae is a Type I DNA/RNA helicase. Mutations in the helicase domain perturb accumulation of diverse RNA classes, and Sen1p has been implicated in 3' end formation of non-coding RNAs. Using a combination of global and candidate-specific two hybrid screens, eight proteins were identified that interact with Sen1p. Interactions with three of the proteins were analyzed further: Rpo21p(Rpb1p), a subunit of RNA polymerase II, Rad2p, a deoxyribonuclease required in DNA repair, and Rnt1p (RNase III), an endoribonuclease required for RNA maturation. For all three interactions, the two hybrid results were confirmed by co-immunoprecipitation experiments. Genetic tests designed to assess the biological significance of the interactions indicate that Sen1p plays functionally significant roles in transcription and transcription-coupled DNA repair. To investigate the potential role of Sen1p in RNA processing and to assess the functional significance of the Sen1p/Rnt1p interaction, we examined U5 snRNA biogenesis. We provide evidence that Sen1p functions in concert with Rnt1p and the exosome at a late step in 3' end formation of one of the two mature forms of U5 snRNA but not the other. The protein-protein and protein-RNA interactions reported here suggest that the DNA/RNA helicase activity of Sen1p is utilized for several different purposes in multiple gene expression pathways. PMID- 15121902 TI - Sequence-structure-function studies of tRNA:m5C methyltransferase Trm4p and its relationship to DNA:m5C and RNA:m5U methyltransferases. AB - Three types of methyltransferases (MTases) generate 5-methylpyrimidine in nucleic acids, forming m5U in RNA, m5C in RNA and m5C in DNA. The DNA:m5C MTases have been extensively studied by crystallographic, biophysical, biochemical and computational methods. On the other hand, the sequence-structure-function relationships of RNA:m5C MTases remain obscure, as do the potential evolutionary relationships between the three types of 5-methylpyrimidine-generating enzymes. Sequence analyses and homology modeling of the yeast tRNA:m5C MTase Trm4p (also called Ncl1p) provided a structural and evolutionary platform for identification of catalytic residues and modeling of the architecture of the RNA:m5C MTase active site. The analysis led to the identification of two invariant residues that are important for Trm4p activity in addition to the conserved Cys residues in motif IV and motif VI that were previously found to be critical. The newly identified residues include a Lys residue in motif I and an Asp in motif IV. A conserved Gln found in motif X was found to be dispensable for MTase activity. Locations of essential residues in the model of Trm4p are in very good agreement with the X-ray structure of an RNA:m5C MTase homolog PH1374. Theoretical and experimental analyses revealed that RNA:m5C MTases share a number of features with either RNA:m5U MTases or DNA:m5C MTases, which suggested a tentative phylogenetic model of relationships between these three classes of 5 methylpyrimidine MTases. We infer that RNA:m5C MTases evolved from RNA:m5U MTases by acquiring an additional Cys residue in motif IV, which was adapted to function as the nucleophilic catalyst only later in DNA:m5C MTases, accompanied by loss of the original Cys from motif VI, transfer of a conserved carboxylate from motif IV to motif VI and sequence permutation. PMID- 15121903 TI - Contact-based sequence alignment. AB - This paper introduces the novel method of contact-based protein sequence alignment, where structural information in the form of contact mutation probabilities is incorporated into an alignment routine using contact-mutation matrices (CAO: Contact Accepted mutatiOn). The contact-based alignment routine optimizes the score of matched contacts, which involves four (two per contact) instead of two residues per match in pairwise alignments. The first contact refers to a real side-chain contact in a template sequence with known structure, and the second contact is the equivalent putative contact of a homologous query sequence with unknown structure. An algorithm has been devised to perform a pairwise sequence alignment based on contact information. The contact scores were combined with PAM-type (Point Accepted Mutation) substitution scores after parameterization of gap penalties and score weights by means of a genetic algorithm. We show that owing to the structural information contained in the CAO matrices, significantly improved alignments of distantly related sequences can be obtained. This has allowed us to annotate eight putative Drosophila IGF sequences. Contact-based sequence alignment should therefore prove useful in comparative modelling and fold recognition. PMID- 15121904 TI - NMR structure of the DNA decamer duplex containing double T*G mismatches of cis syn cyclobutane pyrimidine dimer: implications for DNA damage recognition by the XPC-hHR23B complex. AB - The cis-syn cyclobutane pyrimidine dimer (CPD) is a cytotoxic, mutagenic and carcinogenic DNA photoproduct and is repaired by the nucleotide excision repair (NER) pathway in mammalian cells. The XPC-hHR23B complex as the initiator of global genomic NER binds to sites of certain kinds of DNA damage. Although CPDs are rarely recognized by the XPC-hHR23B complex, the presence of mismatched bases opposite a CPD significantly increased the binding affinity of the XPC-hHR23B complex to the CPD. In order to decipher the properties of the DNA structures that determine the binding affinity for XPC-hHR23B to DNA, we carried out structural analyses of the various types of CPDs by NMR spectroscopy. The DNA duplex which contains a single 3' T*G wobble pair in a CPD (CPD/GA duplex) induces little conformational distortion. However, severe distortion of the helical conformation occurs when a CPD contains double T*G wobble pairs (CPD/GG duplex) even though the T residues of the CPD form stable hydrogen bonds with the opposite G residues. The helical bending angle of the CPD/GG duplex was larger than those of the CPD/GA duplex and properly matched CPD/AA duplex. The fluctuation of the backbone conformation and significant changes in the widths of the major and minor grooves at the double T*G wobble paired site were also observed in the CPD/GG duplex. These structural features were also found in a duplex that contains the (6-4) adduct, which is efficiently recognized by the XPC hHR23B complex. Thus, we suggest that the unique structural features of the DNA double helix (that is, helical bending, flexible backbone conformation, and significant changes of the major and/or minor grooves) might be important factors in determining the binding affinity of the XPC-hHR23B complex to DNA. PMID- 15121906 TI - Listeriosis. PMID- 15121905 TI - A DNA replication-related element downstream from the initiation site of Drosophila selenophosphate synthetase 2 gene is essential for its transcription. AB - Selenophosphate synthetase catalyzes the synthesis of selenophosphate which is a selenium donor for Sec biosynthesis. In Drosophila melanogaster, there are two types of selenophosphate synthetases designated dSPS1 and dSPS2, where dSPS2 is a selenoprotein. The mechanism of gene expression of dSPS2 as well as other selenoproteins in Drosophila has not been elucidated. Herein, we report an essential regulator system that regulates the transcription of the dSPS2 gene (dsps2). Through deletion/substitution mutagenesis, the downstream DNA replication-related element (DRE) located at +71 has been identified as an essential element for dsps2 promoter activity. Furthermore, double-stranded RNA interference (dsRNAi) experiments were performed to ablate transcription factors such as TBP, TRF1, TRF2 and DREF in Schneider cells. The dsRNAi experiments showed that dsps2 promoter activities in DREF- and TRF2-depleted cells were significantly decreased by 90% and 50%, respectively. However, the depletion of TBP or TRF1 did not affect the expression level of dsps2 even though there is a putative TATA box at -20. These results strongly suggest that the DRE/DREF system controls the basal level of transcription of dsps2 by interacting with TRF2. PMID- 15121907 TI - Consultation with the specialist: respiratory failure in children. PMID- 15121908 TI - Hereditary spherocytosis. PMID- 15121909 TI - Vegan diets in infants, children, and adolescents. PMID- 15121910 TI - Index of suspicion. PMID- 15121911 TI - Visual diagnosis: an infant who has a red papule on a swollen, tender arm. PMID- 15121914 TI - History of the medical home concept. PMID- 15121915 TI - The "every child deserves a medical home" training program: more than a traditional continuing medical education course. AB - OBJECTIVES: 1) To develop and implement an innovative, interactive, and nontraditional continuing medical education (CME) curriculum to educate primary care physicians, pediatric office staff, child health advocates, allied health care professionals, and parents of children with special health care needs (CSHCN) about the medical home concept of care and 2) to identify key partners in communities to plan the CME program and ultimately plan for fostering medical homes at the community and state levels. METHODS: Participant outcomes for the CME program and planning process include 1) explaining the elements of the medical home concept as applied to their practice environment or child's care; 2) understanding the concepts, skills, and information necessary to care successfully for CSHCN who are enrolled in managed care organizations; 3) accurately describing trends and developments in caring for CSHCN; 4) identifying programs in the community that serve CSHCN; and 5) assessing and, if necessary, improving pediatric office practices to ensure that they are sensitive to families of children and youths with special health care needs. CONCLUSION: A diverse national committee that included physicians, nonphysicians, and family members developed the Medical Home Training Program curriculum. The medical home curriculum was written to meet the needs of the local community. The training program can offer CME credit and use a direct, outcome-based adult learning technique (eg, determine short- and long-term goals). Furthermore, the program parallels and complements the Healthy People 2010 goals and objectives. PMID- 15121916 TI - Access to the medical home: results of the National Survey of Children with Special Health Care Needs. AB - OBJECTIVE: The purpose of this article is to report the findings of the National Survey of Children With Special Health Care Needs regarding parent perceptions of the extent to which children with special health care needs (CSHCN) have access to a medical home. METHODS: Five criteria, selected to reflect the characteristics of a medical home as defined by the American Academy of Pediatrics (AAP) policy statement on the medical home, were analyzed to describe the extent to which CSHCN receive care characteristic of the medical home concept. These criteria included having 1) a usual place for sick/well care, 2) a personal doctor or nurse, 3) no difficulty in obtaining needed referrals, 4) needed care coordination, and 5) family-centered care received. Items from the Survey were selected and clustered to characterize each of the 5 components. Criteria for each item were established with the requirement that the criteria must be met for all items in a component to receive credit for the component. RESULTS: Results of the survey indicate that 1) approximately half of CSHCN receive care that meets all 5 components established for medical home; 2) most CSHCN have a usual source of care and a personal doctor or nurse, but other components of the medical home, especially elements of care coordination and family-centered care, are lacking; 3) access to a medical home is significantly affected by race/ethnicity, poverty, and the limitations imposed on daily activity by the child's special health care need; and 4) parents of children who do have a medical home report significantly less delayed or forgone care, significantly fewer unmet health care needs, and significantly fewer unmet needs for family support services. The 5 components described represent major characteristics of the comprehensive care model recommended for all children by the AAP. CONCLUSIONS: The findings suggest that although some components of the medical home concept have been achieved for most CSHCN, the comprehensive care model described by the AAP policy statement on the medical home is not yet in place for a significant number of CSHCN and their families. PMID- 15121917 TI - The medical home, access to care, and insurance: a review of evidence. AB - OBJECTIVE: To review the extent to which the literature supports the position that a medical home is important and to review the extent to which insurance is related to having a medical home. METHODS: A review of literature concerning the benefits of a medical home on effectiveness, costs, and equity (reducing disparities) was conducted. RESULTS: International and within-nation studies indicate that a relationship with a medical home is associated with better health, on both the individual and population levels, with lower overall costs of care and with reductions in disparities in health between socially disadvantaged subpopulations and more socially advantaged populations. Although important in facilitating use overall, insurance does not guarantee a medical home. CONCLUSIONS: A medical home, with its 4 key features, provides better effectiveness as well as more efficient and more equitable care to individuals and populations. A concerted attempt to provide a means of universal financial access as well as a medical home should be of high priority for the United States. PMID- 15121918 TI - Building medical homes: improvement strategies in primary care for children with special health care needs. AB - Families and professionals agree that children and adolescents need access to community-based medical homes. This is especially true for children with special health care needs (CSHCN). Most primary care practices are designed for children's routine preventive and acute care needs. CSHCN benefit from care that is integrated with well-child and acute care; coordinated and actively co-managed with specialists, therapists, and educators; and offered in a planned, anticipatory manner. As the primary caregiver and decision maker, families need to be supported in a culturally effective way. Families want community resources that are integrated with care processes. Primary care practices that serve CSHCN require a practical and effective improvement method to become fully realized medical homes. A change method, which blends improvement strategies with progressive measurement, must be perceived by practices as practical and helpful, and it must be supported. The Center for Medical Home Improvement has developed and tested an effective model of medical home improvement and tools to measure the status of a practice and its progress of change. PMID- 15121919 TI - The Pediatric Alliance for Coordinated Care: evaluation of a medical home model. AB - OBJECTIVES: The American Academy of Pediatrics recommends a medical home for children with special health care needs (CSHCN). In the Pediatric Alliance for Coordinated Care (PACC), 6 pediatric practices introduced interventions to operationalize the medical home for CSHCN. The intervention consisted of a designated pediatric nurse practitioner acting as case manager, a local parent consultant for each practice, the development of an individualized health plan for each patient, and continuing medical education for health care professionals. The objectives of this study were 1) to characterize CSHCN in the PACC, 2) to assess parental satisfaction with the PACC intervention, 3) to assess the impact on hospitalizations and emergency department episodes, and 4) to assess the impact on parental workdays lost and children's school days lost for CSHCN before and during the PACC intervention. METHODS: A total of 150 CSHCN in 6 pediatric practices in the Boston, Massachusetts, area were studied. Participants were recruited by their pediatricians on the basis of medical/developmental complexity. Physicians completed enrollment information about each child's diagnosis and severity of condition. Families completed surveys at baseline and follow-up (at 2 years), assessing their experience with health care for their children. RESULTS: A total of 60% of the children had >5 conditions, 41% were dependent on medical technology, and 47% were rated by their physician as having a "severe" condition. A total of 117 (78%) families provided data after the intervention. The PACC made care delivery easier, including having the same nurse to talk to (68%), getting letters of medical necessity (67%), getting resources (60%), getting telephone calls returned (61%), getting early medical care when the child is sick (61%), communicating with the child's doctor (61%), getting referrals to specialists (61%), getting prescriptions filled (56%), getting appointments (61%), setting goals for the child (52%), understanding the child's medical condition (56%), and relationship with the child's doctor (58%). Families of children who were rated "severe" were most likely to find these aspects of care "much easier" with the help of the pediatric nurse practitioner. Satisfaction with primary care delivery was high at baseline and remained high throughout the study. There was a statistically significant decrease in parents missing >20 days of work (26% at baseline; 14.1% after PACC) and in hospitalizations (58% at baseline; 43.2% after PACC). The approximate cost per child per year of the intervention was 400 dollars. CONCLUSIONS: The PACC medical home intervention increases parent satisfaction with pediatric primary care. Those whose needs are most severe seem to benefit most from the intervention. There are some indications of improved health as well as decreased burden of disease with the intervention in place. The PACC model allows a practice to meet many of the goals of serving as a medical home with a relatively small financial investment. PMID- 15121920 TI - Care coordination services in pediatric practices. AB - OBJECTIVE: To examine the frequency with which pediatricians provide care coordination services to children, particularly those with special health care needs, in their practices and the barriers to providing these services. METHODS: An 8-page questionnaire was mailed to 1632 randomly selected US members of the American Academy of Pediatrics. RESULTS: The response rate was 56.7%. Most pediatricians (71.2%) reported that they or someone in their practice serves as the primary care coordinator for their children with special needs, but fewer than one fourth (23.3%) always contact the school about the child's health and educational needs as part of care coordination, only 18.7% always schedule time with the child's family to discuss the findings of a specialist, and only 23.2% meet with the discharge planning team to facilitate transition from hospital to home. The respondents identified the 2 top barriers that impede this activity: limited time and lack of medical staff in their offices. CONCLUSIONS: Although most pediatricians believe that they are providing care coordination services, when asked about specific care coordination activities, such as contacting the school or scheduling time with the family to discuss the findings of a specialist, many do not provide these services. PMID- 15121921 TI - Providing a medical home: the cost of care coordination services in a community based, general pediatric practice. AB - OBJECTIVE: To determine the cost of unreimbursable care coordination services for children with special health care needs (CSHCN) in 1 community-based, general pediatric practice. METHODS: A measurement tool was developed to quantify the precise activities involved in providing comprehensive, coordinated care for CSHCN. Costs of providing this care were calculated on the basis of time spent multiplied by the average salary of the office personnel performing the care coordination service. In addition, data were collected regarding the complexity level of the patient requiring the service, the type of service provided, and the outcome. RESULTS: During the 95-day study period, 774 encounters that led to care coordination activities were logged, representing service provision to 444 separate patients. When these encounters were examined on the basis of clinical complexity of the patient, the most complex patients constituted 11% of the population of CSHCN yet accounted for 25% of the encounters. In addition, care coordination activities for these clinically complex CSHCN engaged office staff 4 times as long when compared with less clinically complex CSHCN. Overall, 51% of the encounters were attributable to coordinating care for problems not considered typically medical and included activities such as processing referrals with managed care organizations, consulting with schools or other educational programs, and providing oversight for psychosocial issues. On the basis of national salary and benefits data, the annual cost of the time spent coordinating care for CSHCN in this medical home model ranged from 22,809 dollars to 33,048 dollars (representing the 25th and 75th percentiles, respectively). CONCLUSIONS: The costs of providing care coordination services to CSHCN in a medical home are appreciable but not prohibitive. Standardization of care coordination practices is essential because it makes the medical home more amenable to quality improvement interventions. Mechanisms to finance unreimbursable care coordination activities must be developed to achieve the Healthy People 2010 objective that all CSHCN have access to a medical home. PMID- 15121922 TI - Using existing population-based data sets to measure the American Academy of Pediatrics definition of medical home for all children and children with special health care needs. AB - OBJECTIVE: National health goals include ensuring that all children have a medical home. Historically, medical home has been determined by the presence of a usual or primary source of care, such as a pediatrician or a family physician. More recent definitions expand on this simplistic notion of medical home. A definition of medical home set forth by the American Academy of Pediatrics (AAP) includes 7 dimensions and 37 discrete concepts for determining the presence of a medical home for a child. Standardized methods to operationalize these definitions for purposes of national, state, health plan, or medical practice level reporting on the presence of medical homes for children are essential to assessing and improving health care system performance in this area. The objective of this study was to identify methods to measure the presence of medical homes for all children and for children with special health care needs (CSHCN) using existing population-based data sets. METHODS: Methods were developed for using existing population-based data sets to assess the presence of medical homes, as defined by the AAP, for children with and without special health care needs. Data sets evaluated included the National Survey of Children With Special Health Care Needs, the National Medical Expenditures Panel Survey, the Consumer Assessment of Health Plans Study Child Survey (CAHPS), and the Consumer Assessment of Health Plans Study Child Survey--Children With Chronic Conditions (CAHPS-CCC2.0H). Alternative methods for constructing measures using existing data were compared and results used to inform the design of a new method for use in the upcoming National Survey of Children's Health. Data from CAHPS CCC2.0H are used to illustrate measurement options and variations in the overall presence of medical homes for children across managed health care plans as well as to evaluate in which areas of the AAP definition of medical home improvements may be most needed for all CSHCN. RESULTS: Existing surveys vary in their coverage of concepts included in the AAP definition of medical home and, therefore, in their capacity to evaluate medical home for children with and without special health care needs. Using data from CAHPS-CCC2.0H, the overall proportion of children who were enrolled in managed care health plans and met criteria for having a medical home varied from 43.9% to 74% depending on the specific scoring method selected for these items. Wide variations across health plans were observed and were most prominent in the areas of "accessible care" and "comprehensive care." Performance was uniformly poorest in the area of "coordinated care" and for CSHCN. Although children with a personal doctor or nurse were more likely to meet the AAP criteria for having a medical home, simply having a personal doctor or nurse was not highly predictive of whether a child experienced the other core qualities of a medical home (positive predictive value: .50; negative predictive value: .59). CONCLUSIONS: Despite differences across existing surveys and gaps in concepts represented, we believe that the AAP definition of medical home can be well represented by the small subset of concepts represented in the National Survey of Children With Special Health Care Needs and the CAHPS-CCC2.0H. A less comprehensive yet still worthwhile measure is possible using the Medical Expenditures Panel Survey. The varying degrees of empirical evidence and consensus for each of the AAP definition domains for medical home suggest the need for constructing measures that also vary in terms of criteria for determining that a child does or does not have a medical home. In addition to a simple "yes or no," or rate-based, measure, a continuous medical "homeness" score that places a child or group of children on a continuum of medical "homeness" is also valuable. Findings indicate that health plans have an important role to play in ensuring medical homes for children in addition to medical practices and those who set policies that guide the design and delivery of health care for children. Oven. Overall, using existing population-based data, a measure of medical home that is aligned with the AAP definition is feasible to include in the annual National Healthcare Quality Report, in state reports on the quality of Medicaid, State Children's Health Insurance Program, and Title V programs as well as to evaluate performance on the Healthy People 2010 objectives and the President's New Freedom Initiative. PMID- 15121923 TI - Implementing community-based systems of services for children and youths with special health care needs: how well are we doing? AB - OBJECTIVE: To provide a baseline measure of the proportion of US children who meet the Maternal and Child Health Bureau's core outcomes for children with special health care needs (CSHCN). Those core outcomes include the following: 1) families of CSHCN will partner in decision making and will be satisfied with the services that they receive; 2) CSHCN will receive coordinated, ongoing comprehensive care within a medical home; 3) families of CSHCN will have adequate private and/or public insurance to pay for the services that they need; 4) children will be screened early and continuously for special health care needs; 5) community-based service systems will be organized so that families can use them easily; and 6) youths with special health care needs will receive the services necessary to make transitions to adult life, including adult health care, work, and independence. METHODS: A national household survey was conducted using telephone interviews. We analyzed data on 38,866 CSHCN included in the 2001 National Survey of CSHCN and 13,579 children included in the 2001 National Health Interview Survey. We assessed the proportion of US children who met each of the 6 core outcomes for CSHCN using data from 2 surveys. RESULTS: Success rates ranged from 6% (the core outcome on successful transition to adulthood) to 74% (the core outcome on organization of the service system). For 5 of the 6 core outcomes, success rates exceeded 50%. CONCLUSION: Our results indicate that, for the most part, the United States is well positioned to meet the 6 core outcomes. However, much more work lies ahead before success can be claimed. This is especially true for the core outcome on transition to adulthood, for which only 6% of children in the target population are now meeting this goal. PMID- 15121924 TI - Policy statement: organizational principles to guide and define the child health care system and/or improve the health of all children. AB - The American Academy of Pediatrics proposed a definition of the medical home in a 1992 policy statement. Efforts to establish medical homes for all children have encountered many challenges, including the existence of multiple interpretations of the "medical home" concept and the lack of adequate reimbursement for services provided by physicians caring for children in a medical home. This new policy statement contains an expanded and more comprehensive interpretation of the concept and an operational definition of the medical home. PMID- 15121926 TI - Are complete blood cell counts useful in the evaluation of asymptomatic neonates exposed to suspected chorioamnionitis? AB - OBJECTIVE: Chorioamnionitis complicates 1% to 10% of pregnancies and increases the risk of neonatal infection. Women with chorioamnionitis receive intrapartum antibiotics, often resulting in inconclusive neonatal blood cultures. Peripheral neutrophil values are used frequently to assist in the diagnosis of neonatal infection and to determine duration of antibiotics; we sought to determine the utility of this approach. METHODS: A prospective observational study was performed in 856 near-term/term neonates who were exposed to suspected chorioamnionitis. Each received antibiotics for 48 hours unless clinical infection or positive blood cultures occurred. Peripheral neutrophils were measured serially and analyzed using the reference ranges of Manroe et al; an additional analysis of only the initial neutrophil values used the normal ranges of Schelonka et al. Results of neutrophil analyses were not used to determine duration of therapy. Fifty percent of asymptomatic neonates were seen postdischarge to ascertain recurrent infection. Local patient charges were examined. RESULTS: Ninety-six percent of neonates were asymptomatic and had negative cultures, and antibiotics were discontinued at 48 hours. A total of 2427 neutrophil counts were analyzed. Although abnormal neutrophil values were more frequent in infected or symptomatic neonates, 99% of asymptomatic neonates had > or = 1 abnormal value. The specificity and negative predictive values for abnormal neutrophil values ranged between 0.12 and 0.95 and 0.91 and 0.97, respectively; sensitivity was 0.27 to 0.76. Significant differences in interpretation of the initial neutrophil values were noted, depending on the normal values used. Follow-up was performed for 373 asymptomatic neonates until 3 weeks' postnatal age. Eight required rehospitalization; none had evidence of bacterial infection. If neutrophil values had been used to determine duration of antibiotics, then local costs would have increased by 76,000 dollars to 425,000 dollars per year. CONCLUSIONS: Single or serial neutrophil values do not assist in the diagnosis of early-onset infection or determination of duration of antibiotic therapy in asymptomatic, culture-negative neonates who are > or = 35 weeks' gestation and are delivered of women with suspected chorioamnionitis. PMID- 15121927 TI - To tap or not to tap: high likelihood of meningitis without sepsis among very low birth weight infants. AB - CONTEXT: Neonatal meningitis is associated with significant morbidity and mortality. We speculated that meningitis may be underdiagnosed among very low birth weight (VLBW) infants because of the failure to perform lumbar punctures (LPs) in infants with suspected sepsis. OBJECTIVE: This study was undertaken to review the epidemiology of late-onset meningitis in VLBW (401-1500 g) infants and to evaluate the concordance of cerebrospinal fluid (CSF) and blood culture (BC) results. METHODS: VLBW infants (excluding those with intraventricular shunts) born at centers of the National Institute of Child Health and Human Development Neonatal Research Network from September 1, 1998, through December 31, 2001, were studied. Late-onset meningitis was defined by culture-based criteria and classified as meningitis with or without associated sepsis. Unadjusted comparisons were made using chi2 tests and adjusted comparisons using regression models. RESULTS: Of 9641 VLBW infants who survived >3 days, 2877 (30%) had > or = 1 LPs, and 6056 (63%) had > or = 1 BC performed after day 3. One hundred thirty four infants had late-onset meningitis (1.4% of all patients; 5% of those with an LP). Pathogens associated with meningitis were similar to those associated with sepsis. One third (45 of 134) of the infants with meningitis had negative BCs. Lower gestational age and prior sepsis increased risk for meningitis. Compared with uninfected infants, those with meningitis had a longer time on mechanical ventilation (28 vs 18 days), had longer hospitalizations (91 vs 79 days), were more likely to have seizures (25% vs 2%), and were more likely to die (23% vs 2%). CONCLUSIONS: Meningitis is a serious complication among VLBW infants, associated with increased severity of illness and risk of death. Of note, one third of the infants with meningitis had meningitis in the absence of sepsis. Because CSF cultures were performed only half as often as BCs, this discordance in blood and CSF culture results suggests that meningitis may be underdiagnosed among VLBW infants. PMID- 15121928 TI - Associations between overweight and obesity with bullying behaviors in school aged children. AB - OBJECTIVE: The prevalence of overweight and obesity in children is rising. Childhood obesity is associated with many negative social and psychological ramifications such as peer aggression. However, the relationship between overweight and obesity status with different forms of bullying behaviors remains unclear. The purpose of this article is to examine these relationships. METHODS: We examined associations between bullying behaviors (physical, verbal, relational, and sexual harassment) with overweight and obesity status in a representative sample of 5749 boys and girls (11-16 years old). The results were based on the Canadian records from the 2001/2002 World Health Organization Health Behaviour in School-Aged Children Survey. Body mass index (BMI) and bullying behaviors were determined from self-reports. RESULTS: With the exception of 15- to 16-year-old boys, relationships were observed between BMI category and peer victimization, such that overweight and obese youth were at greater relative odds of being victims of aggression than normal-weight youth. Strong and significant associations were seen for relational (eg, withdrawing friendship or spreading rumors or lies) and overt (eg, name-calling or teasing or hitting, kicking, or pushing) victimization but not for sexual harassment. Independent of gender, there were no associations between BMI category and bully-perpetrating in 11- to 14-year-olds. However, there were relationships between BMI category and bully perpetrating in 15- to 16-year-old boys and girls such that the overweight and obese 15- to 16-year-olds were more likely to perpetrate bullying than their normal-weight classmates. Associations were seen for relational (boys only) and overt (both genders) forms of bully-perpetrating but not for sexual harassment. CONCLUSIONS: Overweight and obese school-aged children are more likely to be the victims and perpetrators of bullying behaviors than their normal-weight peers. These tendencies may hinder the short- and long-term social and psychological development of overweight and obese youth. PMID- 15121929 TI - The effect of prophylactic ointment therapy on nosocomial sepsis rates and skin integrity in infants with birth weights of 501 to 1000 g. AB - OBJECTIVE: Extremely low birth weight infants have a high risk of developing nosocomial bacterial sepsis (NBS). Immature fragile skin may represent an inadequate protective barrier to bacteria colonizing the skin. We conducted a randomized, multicenter trial to determine whether prophylactic application of an emollient ointment would result in a lower incidence of death and/or NBS in the first 28 days of life, compared with routine skin care. METHODS: Infants of birth weight 501 to 1000 g and gestational age < or =30 weeks were assigned randomly to receive generalized application of ointment twice a day through day 14 (prophylactic group [P]) or local application of ointment to the site of injury (routine skin care [R]). The study was conducted at 53 neonatal intensive care units that were members of the Vermont Oxford Network. RESULTS: Included in the analysis were 1191 infants (P: 602; R: 589). No difference was found in the combined primary outcome of NBS or death (33.6% P vs 30.3% R; relative risk [RR]: 1.10; 95% confidence interval [CI]: 0.89, 1.27). The incidence of death was no different between the groups (10.8% P vs 12.1% R; RR: 0.87; 95% CI: 0.59, 1.25). More infants in the prophylactic group had NBS (25.8% P vs 20.4% R; RR: 1.26; 95% CI: 1.02, 1.54), predominantly in the lower birth weight infants (501-750 g) and for infections caused by coagulase-negative staphylococci. Infants in the prophylactic group had better skin condition on days 1 to 14 of life and less skin injury on days 15 to 28 of life. There was no difference between groups in other complications of prematurity. CONCLUSIONS: Prophylactic application of ointment did not lead to a difference in death and/or NBS in the first 28 days of life. There may be an increase in the risk of NBS associated with this practice. PMID- 15121930 TI - Sleep deprivation for pediatric sedated procedures: not worth the effort. AB - OBJECTIVE: Sleep deprivation is commonly used to enhance the effectiveness of pediatric sedation and to decrease sedation failures. We reviewed our sedation database to evaluate the efficacy of sleep deprivation. METHODS: The entire pediatric sedation unit database (n = 5640) was reviewed retrospectively. Patients without complete data sets were excluded. The remaining patients were separated into 2 groups: 3272 patients who underwent noninvasive procedures and 1210 who underwent invasive procedures. A subgroup of noninvasive procedure patients <2 years old (n = 1398) was also analyzed. The sedation failure rate (%) and nursing care hours for both sleep-deprived and non-sleep-deprived patients were analyzed. RESULTS: In the noninvasive procedure group, the sedation failure rate was 5.7% for the sleep-deprived patients and 5.6% for the non-sleep-deprived patients, whereas the sedation failure rate for children <2 years old was 4.2% for sleep-deprived patients and 4.7% for non-sleep-deprived patients. The sedation failure rate in the invasive procedure group was 7.5% for sleep-deprived patients and 7.2% for non-sleep-deprived patients. Nursing care hours in the noninvasive procedure group were significantly longer for the sleep-deprived patients (4.5 +/- 1.6 hours) versus the non-sleep-deprived patients (3.8 +/- 1.6 hours). This finding was true also for the subgroup of children <2 years old (sleep-deprived patients: 4.2 +/- 1.4 hours; non-sleep-deprived patients: 3.5 +/- 1.4 hours). No difference was noted in nursing care hours for the invasive procedure group. CONCLUSIONS: Sleep deprivation had no effect in reducing the pediatric sedation failure rate. The patients having noninvasive procedures who were sleep deprived required significantly more nursing care hours than their non sleep-deprived counterparts. Routine use of sleep deprivation for pediatric sedation should be critically reevaluated. PMID- 15121931 TI - Parenteral glutamine supplementation does not reduce the risk of mortality or late-onset sepsis in extremely low birth weight infants. AB - BACKGROUND: Glutamine is one of the most abundant amino acids in both plasma and human milk, yet it is not included in standard intravenous amino acid solutions. Previous studies have suggested that parenteral nutrition (PN) supplemented with glutamine may reduce sepsis and mortality in critically ill adults. Whether glutamine supplementation would provide a similar benefit to extremely low birth weight (ELBW) infants is not known. METHODS: We performed a multicenter, randomized, double-masked, clinical trial to assess the safety and efficacy of early PN supplemented with glutamine in decreasing the risk of death or late onset sepsis in ELBW infants. Infants 401 to 1000 g were randomized within 72 hours of birth to receive either TrophAmine (control) or an isonitrogenous study amino acid solution with 20% glutamine whenever they received PN up to 120 days of age, death, or discharge from the hospital. The primary outcome was death or late-onset sepsis. RESULTS: Of the 721 infants who were assigned to glutamine supplementation, 370 (51%) died or developed late-onset sepsis, as compared with 343 of the 712 infants (48%) assigned to control (relative risk: 1.07; 95% confidence interval: 0.97-1.17). Glutamine had no effect on tolerance of enteral feeds, necrotizing enterocolitis, or growth. No significant adverse events were observed with glutamine supplementation. CONCLUSIONS: Parenteral glutamine supplementation as studied did not decrease mortality or the incidence of late onset sepsis in ELBW infants. Consequently, although no harm was demonstrated, routine use of parenteral glutamine supplementation cannot be recommended in this population. PMID- 15121932 TI - The bedding environment, sleep position, and frequent wheeze in childhood. AB - OBJECTIVE: Synthetic quilt use has been associated with increased childhood wheeze in previous studies. Our aim was to examine whether the adverse effect of synthetic quilt use on frequent wheeze differed by usual sleep position. DESIGN, SETTING, AND PARTICIPANTS: A population-based cross-sectional study of 6378 (92% of those eligible) 7-year-olds in Tasmania, Australia, was conducted in 1995. Exercise-challenge lung function was obtained on a subset of 414 children from randomly selected schools. EXPOSURE MEASURES: Child bedding including pillow and overbedding composition and usual sleep position by parental questionnaire. OUTCOME MEASURES: Frequent wheeze (>12 wheeze episodes over the past year), using the International Study of Asthma and Allergies in Childhood parental questionnaire, and baseline and postexercise forced expiratory volume in 1 second lung-function measures. RESULTS: Frequent wheeze (n = 117) was positively associated with synthetic quilts, synthetic pillows, electric blankets, and sleeping in a bottom bunk bed but did not vary by sleep position. In a nested case-control analysis, the association between synthetic quilt use and frequent wheeze differed by sleep position. Among children who slept supine, synthetic (versus feather) quilt use was associated with frequent wheeze (adjusted odds ratio: 2.37 [1.08, 5.23]). However, among nonsupine sleepers, overlying synthetic quilt use was not associated with frequent wheeze (adjusted odds ratio: 1.06 [0.60, 1.88]). This difference in quilt effect by sleep position was highly significant. Similarly, synthetic quilt use was associated with lower postexercise forced expiratory volume in 1 second measures among supine but not nonsupine sleeping children. CONCLUSION: An increasing focus on the bedding environment immediately adjacent to the nose and mouth is required for respiratory disorders provoked by bedding, such as child asthma characterized by frequent wheeze. PMID- 15121933 TI - Changes in mortality for extremely low birth weight infants in the 1990s: implications for treatment decisions and resource use. AB - OBJECTIVE: Much has changed in neonatal intensive care unit (NICU) care over the past decade. High-frequency oscillation, inhaled nitric oxide, and antenatal corticosteroids are now widely available. We wondered how these medical advances had affected both the epidemiology and ethics of life and death for extremely low birth weight (ELBW) infants in the NICU. METHODS: We identified 1142 ELBW infants (birth weight [BW] < 1000 g) consecutively admitted to our NICU between 1991 and 2001. We abstracted BW, gestational age, survival or death, and length of stay in the NICU. Statistical analyses were performed by using linear regression and 2 way analysis of variance. RESULTS: Both increasing BW and later year were significantly associated with improved survival. However, for larger ELBW infants, survival was approximately 90% for the entire decade, and large-scale improvement was hardly possible. For smaller infants, greater improvements were both possible and observed, at least early in the decade. From 1991 to 1997, overall ELBW survival increased steadily (approximately 4% per year). However, from 1997 to 2001, there was no significant improvement in survival for ELBW infants. There was no change in the distribution of deaths accounted for by BW subgroups within the ELBW population from 1991 to 2001. Median length of stay for infants who eventually expired before discharge rose from 2 days in 1991 to 10 days in 2001. As a consequence, during the past decade, the percentage of infants whose outcome was "undeclared" by day of life 4 rose from 10% to 20% for ELBW infants overall and to 33% for infants with BWs of 450 to 700 g. The percentage of ELBW NICU bed-days occupied by nonsurvivors remained very low (approximately 7%) from 1991 to 2001. CONCLUSIONS: 1) Fewer infants in all ELBW subgroups are dying, compared with a decade ago, and the improvement has been most prominent for BWs of 450 to 700 g, at which mortality was and remains to be greatest. 2) This progress seems to have slowed, or even stopped, by the end of the decade. 3) Although most NICU nonsurvivors still expire early, doomed infants are lingering longer. 4) Nonsurvivors continue to occupy a constant (and extremely small) fraction of NICU bed-days. PMID- 15121934 TI - Preterm delivery, level of care, and infant death in sweden: a population-based study. AB - OBJECTIVE: To elucidate the role of level of care in combination with other perinatal risk factors for infant death in very preterm deliveries. DESIGN: Population-based cohort study. SETTING: Sweden, 1992-1998. SUBJECTS: Singleton infants (2285) born at 24 to 31 completed weeks of gestation to primiparous women. MAIN OUTCOME MEASURE: Infant mortality. RESULTS: The rate of infant mortality increased from 5% among infants born at 31 weeks' gestation to 56% among infants born at 24 weeks' gestation. Compared with infants born at university hospitals, the unadjusted odds ratio (OR) of infant death was 0.70 (95% confidence interval [CI]: 0.54-0.90) among infants delivered at general hospitals. However, after adjustment, the OR of infant death shifted to 1.33 (95% CI: 0.88-2.02) for preterm births at general hospitals. This shift was primarily due to different gestational age distributions in regional and general hospitals. Among infants born at 24 to 27 weeks' gestation, infant mortality rates were 23% (87 deaths) in university hospitals and 32% (73 deaths) in general hospitals, giving an adjusted OR of 2.00 for general versus university hospitals (95% CI: 1.15-3.49). The risk of death at 24 to 27 weeks' gestation in general hospitals was increased specifically in pregnancies with placental complications. CONCLUSION: Taking obstetric complications into account, there is an excess mortality risk among extremely preterm infants born at general hospitals. PMID- 15121935 TI - Discipline in the African American community: the impact of socioeconomic status on beliefs and practices. AB - OBJECTIVE: To describe and compare disciplinary beliefs and practices among African American parents from diverse socioeconomic backgrounds. METHODS: A cross sectional survey was conducted of self-identified African American parents of children <48 months of age at 2 ambulatory teaching clinics, 2 community health centers, and 3 private practices in Washington, DC, and the surrounding metropolitan area. Disciplinary beliefs and practices of African American parents were measured. RESULTS: A total of 175 of the 189 parents who were approached for the study completed the survey for a participation rate of 92.5%. Middle/upper socioeconomic status (SES) parents in this study were more likely to be married (60.9% vs 14.7%), older (31.4 years vs 25 years), and more educated (80% having attended at least some college vs 34.4%) than lower SES parents. There were no significant differences between middle/upper and lower SES parents with regard to their belief in a preferred disciplinary method (teaching, spanking, removing) or approach (positive, negative). Lower SES parents were more likely to endorse spanking a 1- to 3-year-old child if they were doing something that was not safe (90.5% vs 78.3%). Middle/upper SES parents were significantly more likely to reward their child for positive behavior than lower SES parents (66.1% vs 47.1%). CONCLUSIONS: Lower and middle/upper SES parents in this study population were reasonably similar with respect to disciplinary beliefs and practices. Exceptions to this generalization were that lower SES parents were more likely to endorse spanking as a response to an unsafe behavior on the part of the child, and middle/upper SES parents reported higher levels of reward for positive behavior. PMID- 15121936 TI - Postnatal depression and faltering growth: a community study. AB - OBJECTIVE: To investigate the association between faltering growth in children and maternal postnatal depression. METHODS: Children aged < or =2 years were identified from community child health surveillance records if their weights fell across 2 centile channels on standardized growth charts or fell below the second centile. Mothers of these index children were invited to complete the Edinburgh Postnatal Depression Scale and the anxiety subscale of the Hospital Anxiety and Depression Scale. Those who scored above threshold values on either scale were interviewed with the revised Clinical Interview Schedule. Matched control children were obtained from health visitor records, and records of their weights were obtained. Mothers of control children completed the same questionnaires. RESULTS: A total of 196 index children and 567 control children were studied. Significantly more mothers in the index group scored above the threshold for both the Edinburgh Postnatal Depression Scale (33% vs 22%; odds ratio [OR]: 1.71; 95% confidence interval [CI]: 1.16-2.53) and the Hospital Anxiety and Depression Scale (24% vs 13%; OR: 2.08; 95% CI: 1.33-3.25) questionnaires. Furthermore, clinical interviews with these mothers demonstrated that 21% of the index group and 11% of the control group fulfilled criteria for depressive episode (OR: 1.88; 95% CI: 1.21-2.94). CONCLUSIONS: Depression in mothers of children with faltering growth during the first 2 years of life is significantly greater than in mothers of children who are gaining weight appropriately. In view of the high rates of maternal depression in children with poor weight gain, clinical management at presentation of either problem should focus on both members of the mother-child dyad and on the interaction between mother and child. These findings have implications for all professionals who work in primary and secondary health care. PMID- 15121937 TI - More evidence for reach out and read: a home-based study. AB - OBJECTIVE: Reach Out and Read (ROR), a clinic-based literacy program, has been shown to improve literacy outcomes in impoverished children. No study has used direct observation to assess a child's home literacy environment or to control for important confounders, such as the quality of the home environment. The objective of this study was to determine the relationship between the frequency of ROR encounters that a family receives during well-child visits and a child's home literacy profile, while accounting for important confounders, such as the quality of the home environment. METHODS: A cross-sectional study was conducted of 137 children (aged 18-30 months) who received pediatric well-child care at the Yale-New Haven Hospital Primary Care Center. The number of ROR encounters that the family received was determined though parent interview, direct observation, and a review of the medical record. After a brief waiting room interview, a home visit was conducted. An assessment of the child's home literacy environment was completed on the basis of 10 variables that were obtained from parent report and direct observation within the home. These variables were summed to form a Child Home Literacy Index. The Home Observation for Measurement of the Environment, a standardized measure of the nurturing quality of the home environment, was also administered. Hierarchical linear regression was conducted to determine the significance of ROR on a child's home literacy environment. RESULTS: A total of 100 families completed both a waiting room interview and a home visit. Families received between 0 and 6 books in the ROR program. A total of 93% of families reported reading to their children, but only 35% of parents identified reading as a favorite activity of their child and 45% of parents reported that this was a favorite joint activity. Hierarchical linear regression demonstrated that increasing frequency of ROR encounters contributed a small but significant portion of the variance explaining a child's home literacy profile (5%), with this model accounting for a total of 19% of the variance. CONCLUSIONS: A modest literacy intervention, such as ROR, can have a significant impact on a child's home literacy environment. PMID- 15121938 TI - Clinical features of children with screening-identified evidence of celiac disease. AB - OBJECTIVE: At-risk groups commonly undergo screening for autoantibodies associated with celiac disease (CD). However, the clinical significance of a positive test remains uncertain. The objective of this study was to evaluate growth and clinical features of children who test positive for an autoantibody associated with CD. METHODS: A case-control study of Denver area healthy infants and young children with and without CD autoantibodies was conducted. A cohort of HLA-characterized children were followed prospectively since birth for the development of immunoglobulin A antitissue transglutaminase autoantibodies (TG). Clinical evaluation, questionnaire, blood draw, and small bowel biopsy were performed. Growth and nutrition and frequency of positive responses were measured. RESULTS: Compared with 100 age- and gender-matched TG-negative controls, 18 TG-positive children, 5.5 +/- 0.5 years of age, had a greater number of symptoms and lower z scores for weight-for-height and for body mass index. Responses that were independently associated with TG-positive status were irritability/lethargy, abdominal distention/gas, and difficulty with weight gain. CONCLUSIONS: Screening-identified TG-positive children demonstrate mild alterations in growth and nutrition and report more symptoms than control subjects. Additional study is needed on the benefit and risk of identifying CD in at-risk groups. PMID- 15121939 TI - Immune parameters and morbidity in hard drug and human immunodeficiency virus exposed but uninfected infants. AB - OBJECTIVE: To examine the association of maternal hard drug use (injection drugs, cocaine, and opiates) on lymphocyte subsets and clinical morbidity in uninfected infants who are born to human immunodeficiency virus-infected mothers who were enrolled in the Women and Infants Transmission Study (1990-2000). METHODS: Maternal hard drug use was identified by self-report and/or urine toxicology. Infant evaluations occurred at birth and at 1, 2, 4, 6, 9, 12, 18, and 24 months of age. RESULTS: A total of 401 (28%) of the 1436 uninfected infants were born to drug-using mothers. Maternal CD4 lymphocyte percentage and RNA at delivery were not significantly different between drug users and nonusers. Infants who were born to drug-using mothers had lower mean gestational age (37.8 vs 38.5 weeks) and birth weight (2.9 vs 3.1 kg). Infants with intrauterine drug exposure had lower CD4 lymphocyte percentage over the first 4 months of life after adjusting for covariates and higher natural killer lymphocyte percentage. When the analysis was stratified by time period of entry, the incidence of clinical events was not different between infants who were born to drug users versus nonusers. CONCLUSION: Maternal hard drug use is associated with immunologic changes in infants early in life, although these changes did not seem to be associated with increased risk of infections. PMID- 15121940 TI - Low expression of human epithelial sodium channel in airway epithelium of preterm infants with respiratory distress. AB - OBJECTIVE: Active ion transport is critical to postnatal clearance of lung fluid. The importance of epithelial sodium channel (ENaC) in this clearance has been demonstrated in animal studies in which alpha-ENaC knockout mice died postnatally as a result of respiratory insufficiency. In animals, the expression of alpha ENaC in respiratory epithelium is dependent on gestational age, but when assessed by in situ hybridization in the human (h), the mRNA is present from the earliest stages of pulmonary development. Therefore, the purpose of the present investigation was to quantify mRNA of the alpha-, beta-, and gamma-hENaC subunits of newborn preterm infants with respiratory distress and compare the gene expression data against those detected in healthy term infants. In addition, the effect of systemic dexamethasone therapy on the 3 hENaC subunits was studied in 4 preterm infants who received prolonged assisted ventilation. METHODS: The expression of subunits of hENaC was determined in samples taken from nasal respiratory epithelium of 7 healthy term infants (gestation age: 39.3 +/- 0.9 weeks [mean +/- standard deviation) and 5 preterm infants (gestational age: 27.2 +/- 0.9 weeks) with respiratory distress syndrome within 5 hours of birth. Betamethasone had been given to all mothers of preterm infants. In 4 additional preterm infants who still required assisted ventilation at 43 +/- 6 days postnatal age, the expression of alpha-hENaC was determined in samples taken before and during treatment with dexamethasone. RESULTS: Preterm infants with respiratory distress syndrome had low expression of all hENaC subunits relative to healthy term infants (alpha-hENaC: 5.38 +/- 2.01 [amol/fmol cytokeratin 18] vs 9.13 +/- 2.26; beta-hENaC: 2.44 +/- 1.43 vs 4.25 +/- 1.10; gamma-hENaC: 2.43 +/- 0.11 vs 6.81 +/- 3.24). Each of the 4 preterm infants who were treated with dexamethasone at approximately 1 month of age showed an increase in expression of alpha-hENaC and beta-hENaC subunit normalized to cytokeratin 18. CONCLUSION: All 3 subunits of the hENaC are low in preterm relative to full-term infants. alpha hENaC mRNA in respiratory epithelium is increased by therapeutic doses of glucocorticosteroid. Low expression of alpha-hENaC in human respiratory epithelium may play a role in the pathogenesis of respiratory distress in preterm infants. PMID- 15121941 TI - Serum leptin concentration poorly reflects growth and energy and nutrient intake in young children. AB - OBJECTIVE: We studied whether the serum leptin concentration at age 2 years predicts changes in relative body weight by age 8 and whether the serum leptin concentration is associated with intake of energy and nutrients at age 5. METHODS: A total of 156 8-year-old participants of the Special Turku Coronary Risk Factor Intervention Project were chosen to represent children whose relative weight decreased, was stable, or increased during the preceding 6 years. Their serum leptin concentrations were measured in samples collected when they were 2 years. Serum leptin was also measured in 100 5-year-old children in the Special Turku Coronary Risk Factor Intervention Project whose energy and nutrient intakes were analyzed using 4-day food records. RESULTS: The boys whose relative weight decreased (n = 25), was stable (n = 28), or increased (n = 26) between 2 and 8 years of age had similar serum leptin concentrations at the age of 2 years. The girls whose relative weight decreased (n = 27) had higher serum leptin concentrations at 2 years than the girls whose relative weight remained stable (n = 26) but only when the leptin values were not adjusted for body mass index. The serum leptin concentration was higher in 5-year-old girls than in 5-year-old boys even when adjusted for body mass index. Serum leptin correlated with relative weight in girls and boys (r = 0.65 and r = 0.45, respectively). Serum leptin concentration adjusted for relative weight correlated poorly with intakes of energy, fat, saturated fat, carbohydrates, sucrose, and protein. CONCLUSIONS: Serum leptin concentrations at age 2 poorly predicted changes in relative body weight during the following 6 years and poorly reflected the intake of energy or major nutrients at age 5. PMID- 15121942 TI - Acute hyponatremia related to intravenous fluid administration in hospitalized children: an observational study. AB - OBJECTIVE: To develop hyponatremia (plasma sodium concentration [P(Na)] <136 mmol/L), one needs a source of water input and antidiuretic hormone secretion release to diminish its excretion. The administration of hypotonic maintenance fluids is common practice in hospitalized children. The objective of this study was to identify risk factors for the development of hospital-acquired, acute hyponatremia in a tertiary care hospital using a retrospective analysis. METHODS: All children who presented to the emergency department in a 3-month period and had at least 1 P(Na) measured (n = 1586) were evaluated. Those who were admitted were followed for the next 48 hours to identify patients with hospital-acquired hyponatremia. An age- and gender-matched case-control (1:3) analysis was performed with patients who did not become hyponatremic. RESULTS: Hyponatremia (P(Na) <136 mmol/L) was documented in 131 of 1586 patients with > or = 1 P(Na) measurements. Although 96 patients were hyponatremic on presentation, our study group consisted of 40 patients who developed hyponatremia in hospital. The case control study showed that the patients in the hospital-acquired hyponatremia group received significantly more EFW and had a higher positive water balance. With respect to outcomes, 2 patients had major neurologic sequelae and 1 died. CONCLUSION: The most important factor for hospital-acquired hyponatremia is the administration of hypotonic fluid. We suggest that hypotonic fluid not be given to children when they have a P(Na) <138 mmol/L. PMID- 15121943 TI - Measurement of percentage of body fat in 411 children and adolescents: a comparison of dual-energy X-ray absorptiometry with a four-compartment model. AB - OBJECTIVE: Pediatricians are encountering body composition information more frequently, with percentage of body fat (%BF) measurement receiving particular attention as a result of the obesity epidemic. One confounding issue is that different methods may yield different %BF results in the same person. The objective of this study was to compare dual-energy X-ray absorptiometry (DXA) with the criterion 4-compartment model (4-CM) for measurement of %BF in a large pediatric cohort and to assist pediatricians in appropriate interpretation of body composition information by recognizing differences between techniques. METHODS: Height, weight, anthropometrics, body density by underwater weighing, total body water by deuterium dilution, and bone mineral content and %BF by DXA (Lunar DPX/DPX-L) were measured in 411 healthy subjects, aged 6 to 18 years. Values for %BF by 4-CM and DXA were compared using regression analysis. RESULTS: The mean +/- standard deviation values for %BF by DXA (22.73% +/- 11.23%) and by 4-CM (21.72% +/- 9.42%) were different, but there was a strong relationship between the 2 methods (R2 = 0.85). DXA underestimated %BF in subjects with lower %BF and overestimated it in those with higher %BF. The relationship between the 2 methods was not affected by gender, age, ethnicity, pubertal stage, height, weight, or body mass index. The standard error of the estimate was 3.66%. CONCLUSION: This analysis demonstrates a predictable relationship between DXA and 4-CM for %BF measurement. Because of its ease of use, consistent relationship with 4-CM, and availability, we propose that DXA has the capacity for clinical application including prediction of metabolic abnormalities associated with excess %BF in pediatrics. PMID- 15121944 TI - A cost-effectiveness analysis of bacterial endocarditis prophylaxis for febrile children who have cardiac lesions and undergo urinary catheterization in the emergency department. AB - OBJECTIVE: To prevent bacterial endocarditis (BE) in those at risk, the American Heart Association recommends antibiotics for patients who have a known urinary tract infection and are about to undergo urinary catheterization (UC). In young children who have cardiac lesions and undergo UC for fever without a source, the problem with prophylaxis only in the presence of infected urine is that the presence of urinary tract infection is unknown before testing. This study was conducted to determine the cost-effectiveness of BE prophylaxis before UC in febrile children aged 0-24 months with moderate-risk cardiac lesions. METHODS: We evaluated the cost-effectiveness of BE prophylaxis compared with no prophylaxis from the societal perspective. Clinical outcomes were based on BE incidence and quality-adjusted life years (QALYs). Probabilities were derived from the medical literature. Costs were derived from national and local sources in US dollars for the reference year 2000, using a discount rate of 3%. RESULTS: On the basis of the analysis, prophylaxis prevents 7 BE cases per 1 million children treated. When antibiotic-associated deaths were included, the no-prophylaxis strategy was more effective and less costly than the prophylaxis strategy. When antibiotic associated deaths were excluded, amoxicillin cost 10 million dollars per QALY gained and 70 million dollars per case prevented. For vancomycin, it was 13 million dollars per QALY gained and 95 million dollars per case prevented. The results were robust to variations in the prophylactic efficacy of antibiotics, incidence of bacteremia after UC, incidence of BE after bacteremia, and costs associated with BE prophylaxis and treatment. CONCLUSION: In the emergency department, BE prophylaxis before UC in febrile children who are aged 0 to 24 months and have moderate-risk cardiac lesions is not a cost-effective use of health care resources. PMID- 15121945 TI - Cognitive and motor development among small-for-gestational-age infants: impact of zinc supplementation, birth weight, and caregiving practices. AB - OBJECTIVE: Infants who are born small for gestational age (SGA) are at risk for developmental delays, which may be related to deficiencies in zinc, an essential trace metal, or to deficiencies in their ability to elicit caregiver responsiveness (functional isolation hypothesis). The objective of this study was to evaluate at 6 and 10 months of age the impact of a 9-month supplementation trial of 5 mg of zinc on the development and behavior of infants who were born SGA and to evaluate infants' ability to elicit responsive caregiver behavior. METHODS: A randomized, controlled trial of zinc supplementation was conducted among 200 infants in a low-income, urban community in Delhi, India. Infants were recruited when they were full term (>36 weeks) and SGA (birth weight <10th percentile weight-for-gestational age). Infants were randomized to receive daily supplements of a micronutrient mix (folate, iron, calcium, phosphorus, and riboflavin) with or without 5 mg of zinc sulfate. The supplement was administered by field workers daily from 30 days to 9 months of age. At 6 and 10 months, infant development and behavior were measured in a clinical setting using the Bayley Scales of Infant Development II. Caregiver responsiveness, observed on an Indian version of the Home Observation for Measurement of the Environment scale, was measured during a home visit at 10 months. During both the clinic and home visits, caregivers reported on their infant's temperament. RESULTS: There were no direct effects of zinc supplementation on the infants' development or behavior at either 6 or 10 months. In a subgroup analysis among the zinc-supplemented infants, lower birth weight infants were perceived to be more temperamentally difficult than higher weight infants; in the control group, birth weight was not associated with temperament. Heavier birth weight infants had better scores on all measures of development and behavior at 6 months and on changes in mental and motor development from 6 to 10 months, compared with lighter birth weight infants. Boys had better weight gain and higher scores on mental development and emotional regulation than girls. Infants who were from families of higher socioeconomic status (indexed by parental education, house size, and home ownership) had higher scores on mental development and orientation/engagement (exploratory behavior) than infants who were from families of lower socioeconomic status. In keeping with the functional isolation hypothesis, caregiver responsiveness was associated with infant irritability, controlling for socioeconomic status, gender, birth weight, and weight gain. Responsive mothers were more likely to perceive their infants to be temperamentally easy than less responsive mothers. CONCLUSION: Possible explanations for the lack of effects of zinc supplementation on infant development and behavior include 1) subtle effects of zinc supplementation that may not have been detected by the Bayley Scales, 2) interference with other nutritional deficiencies, or 3) no impact of zinc deficiency on infants' development and behavior. The link between birth weight and irritability among infants in the zinc supplementation group suggests that the response to zinc supplementation may differ by birth weight, with irritability occurring among the most vulnerable infants. Longer term follow-up studies among zinc-supplemented infants are needed to examine whether early supplementation leads to developmental or behavioral changes that have an impact on school-age performance. The relationship between infant irritability and low maternal responsiveness lends support to the functional isolation hypothesis and the importance of asking caregivers about infant temperament. PMID- 15121946 TI - Outcomes in children and young adults who are hospitalized for firearms-related injuries. AB - OBJECTIVES: To compare outcomes by intent of nonfatal firearms-related injuries in a hospitalized population, newborn to 19 years of age, and estimate the national incidence of ensuing disability. METHODS: Descriptive statistics and comparative analysis using chi(2), odds ratio, and t test were applied to data from the National Pediatric Trauma Registry (NPTR) and the National Electronic Injury Surveillance System of the US Consumer Product Safety Commission. Demographics, preinjury medical history, scene of injury, primary body part injured, severity of injury, utilization of resources, short-term and long-term disability, medical cause of disability, and disposition at discharge were studied. RESULTS: NPTR unintentional (n = 268) and assault-related firearms related injuries (n = 506) were compared. In both groups, the majority of patients were male (80%). Compared with the unintentionally injured, the assaulted children were older and more frequently black (59.3% vs 32.5%). Approximately 17% in both groups had a preinjury history of medical/psychosocial problems. Unintentional injuries occurred mainly in private dwellings (75.7%), and assaults occurred in public places/street (53.8%). In both groups, injuries to multiple body regions were prevalent, and a substantial proportion sustained injuries of serious to critical level. Most children were transported by ambulance, but a significant proportion in the unintentional group were transported by helicopter. The rate of admission to the intensive care unit was approximately 40% for both groups. The unintentionally injured had a higher rate of surgical intervention (66.8% vs 50.8%) and stayed in the hospital longer than the assaulted ones (median: 5 days vs 3 days). Almost half of the children in both groups were discharged with disability, and approximately 87% returned to their home. Applying the NPTR disability rate to National Electronic Injury Surveillance System estimates of hospitalization suggests that approximately 3200 children nationwide develop disability from firearms-related injuries annually. CONCLUSIONS: Nonfatal firearms-related injuries in a pediatric population are associated with a high use of medical resources and lasting disability. Public policies should be developed and implemented to reduce the occurrence of these catastrophic events. PMID- 15121947 TI - Depressive symptoms and health risk among rural adolescents. AB - OBJECTIVE: To determine the stability of depression and its relationship with health risk factors among rural adolescents. METHODS: A clinic-based longitudinal study was conducted to test for depression and risk factors in 64 participants who attended a rural, primary care, adolescent medicine clinic. The primary measure of risk and depression was the Perkins Adolescent Risk Screen (PARS). Adolescent patients who were aged 12 to 18 years and had PARS assessments during a previous visit to the adolescent clinic were invited to complete a follow-up PARS assessment. RESULTS: The mean age of adolescents at baseline was 12.79 years; 14.59 years at follow-up. With age and gender being controlled, adolescent depression and various adolescent risk indices were significantly related at baseline. Longitudinally, baseline depression score on PARS were related to follow-up: depression, school problems, substance abuse, tobacco use, sexual activity, and violent behavior scores and a history of physical/sexual abuse. On multivariate analysis controlling for other significantly associated variables, the relationship persisted for baseline depression and follow-up: tobacco, substance abuse, depression, and history of physical/sexual abuse. CONCLUSION: This study confirms a strong longitudinal relationship between baseline depressive symptoms and several important risk behaviors/factors measured at follow-up in a clinic population of rural adolescents. Also, longitudinal stability of depression over time is supported. PMID- 15121948 TI - Spanking in early childhood and later behavior problems: a prospective study of infants and young toddlers. AB - OBJECTIVE: To explore the relationship of spanking frequency before age 2 with behavior problems near time of entry into school. METHODS: Children who were younger than 2 years were followed up approximately 4 years later, after they had entered school. The likelihood of significant behavior problems at follow-up was estimated in multivariate analyses that controlled for baseline spanking frequency and other characteristics. Participants were mothers from a large-scale national study and their children. Statistical analysis included an ethnically diverse sample of 1966 children aged 0 to 23 months at baseline. Two dichotomous indicators of behavior problems were used. The first indicated that maternal rating of child behavior problems exceeded a threshold. The second indicated that a mother met with a school administrator to discuss her child's behavior problems. RESULTS: White non-Hispanic children who were spanked more frequently before age 2 were substantially more likely to have behavior problems after entry into school, controlling for other factors. For Hispanic and black children, associations between spanking frequency and behavior problems were not statistically significant and were not consistent across outcome measures. CONCLUSION: Among white non-Hispanic children but not among black and Hispanic children, spanking frequency before age 2 is significantly and positively associated with child behavior problems at school age. These findings are consistent with those reported in studies of children older than 2 years but extend these findings to children who are spanked beginning at a relatively early age. PMID- 15121949 TI - Comparison of two educational interventions on pediatric resident auscultation skills. AB - OBJECTIVE: Multiple cross-sectional physician surveys have documented poor cardiac auscultation skills. We evaluated the impact of 2 different educational interventions on pediatric resident auscultation skills. METHODS: The auscultation skills of all first-year (PGY1; n = 20) and second-year pediatric residents (PGY2; n = 20) were evaluated at the beginning and end of the academic year. Five patient recordings were presented: atrial septal defect, ventricular septal defect, pulmonary valve stenosis, bicuspid aortic valve with insufficiency, and innocent murmur. Residents were asked to classify the second heart sound, identify a systolic ejection click, describe the murmur, and provide a diagnosis. All PGY1 and most PGY2 (14 of 20) participated on the inpatient cardiology service for 1 month. PGY2 on the cardiology service also attended outpatient clinic. PGY1 did not attend outpatient clinic but were allotted 2 hours/week to use a self-directed cardiac auscultation computer teaching program. RESULTS: Resident auscultation skills on initial evaluation were dependent on training level (PGY1: 42 +/- 15% correct; PGY2: 53 +/- 13% correct), primarily as a result of better classification of second heart sound (PGY1: 45%; PGY2: 63%) and diagnosis of an innocent murmur (PGY1: 35%; PGY2: 65%). There was no difference in the ability to identify correctly a systolic ejection click (20% vs 23%) or to arrive at the correct diagnosis (35% vs 40%). At the end of the academic year, the PGY1 scores improved by 21%, primarily as a result of improved diagnostic accuracy of the innocent murmur (35% to 65%). PGY2 scores remained unchanged (53% vs 51%), regardless of participation in a cardiology rotation (cardiology rotation: 50%; no cardiology rotation: 51%). Combined, diagnostic accuracy was best for ventricular septal defect (55%) and innocent murmur (60%) and worst for atrial septal defect (18%) and pulmonary valve stenosis (15%). However, 40% identified the innocent murmur as pathologic and 21% of pathologic murmurs were diagnosed as innocent. CONCLUSIONS: Pediatric resident auscultation skills were poor and did not improve after an outpatient cardiology rotation. Auscultation skills did improve after the use of a self-directed cardiac auscultation teaching program. These data have relevance given the American College of Graduate Medical Education's emphasis on measuring educational outcomes and documenting clinical competencies during residency training. PMID- 15121950 TI - Placement stability and mental health costs for children in foster care. AB - OBJECTIVE: Although prior population-based studies have found that children in foster care use more mental health services than their Medicaid peers, less is known about how different experiences in foster care impact the likelihood of mental health service use. The primary aim of this study is to test the hypothesis that instability of foster care placements is associated with higher costs for mental health care services. The secondary aim is to test the hypothesis that foster care children are also more likely to generate high costs for mental health services if they generate higher costs for non-mental health claims. METHODS: Using administrative child welfare data linked to Medicaid claims, we assembled a unique retrospective cohort of adjudicated dependent children >2 years old who entered foster care between July 1993 and June 1995, spent at least 9 months in care, and were Medicaid eligible during a 1-year follow-up period. The primary outcome was high mental health service use, defined as having costs in the top decile of the sample. The primary independent variables were the number of foster care placements during the year and whether placements were interrupted by a return home for at least 1 month during that year (episodic foster care). We used logistic regression to estimate the association between placements and service utilization, with adjustment for age and physical health care costs. RESULTS: Of the 1635 children in the study, 41% had > or = 3 foster care placements, and 5% had episodic foster care during the year of observation. The top 10% of mental health service users accounted for 83% of the 2.4 million dollars in mental health costs. Both multiple placements and episodic foster care increased the predicted probability of high mental health service use. Higher physical health care costs also increased the probability of high mental health use for all children, but this increased probability was most dramatic among children with episodic foster care (probability of high mental health use: 0.78; 95% confidence interval: 0.42-0.94). CONCLUSIONS: Foster care placement instability was associated with increased mental health costs during the first year in foster care, particularly among children with increasing general health care costs. These findings highlight the importance of interventions that address the global health of children in foster care and may permit better targeting of health care resources to subgroups of children most likely to use services. PMID- 15121951 TI - Does pediatric surgical specialty training affect outcome after Ramstedt pyloromyotomy? A population-based study. AB - OBJECTIVE: Ramstedt pyloromyotomy is a common operation in infants and is often done by general surgeons. We wished to determine whether there are any differences in outcome when this procedure is done by subspecialist pediatric general surgeons as compared with general surgeons. METHODS: All Ramstedt pyloromyotomies in the province of Ontario between 1993 and 2000 were reviewed. Children with complex medical conditions or prematurity were excluded. Cases done by general surgeons were compared with those done by pediatric surgeons, specifically examining hospital stay and complications. RESULTS: Of 1777 eligible infants, 67.9% were operated on by pediatric surgeons and 32.1% by general surgeons. Total and postoperative lengths of stay were longer in the general surgeon group compared with the pediatric surgeons (4.31 vs 3.50 days for length of stay; 2.95 vs 2.25 days for postoperative length of stay). The general surgeons had a higher overall complication rate (4.18% vs 2.58%). The incidence of duodenal perforation among general surgeons was almost 4 times that of pediatric surgeons (relative risk: 3.65; 95% confidence interval: 1.43-9.32). Of the 4 infants who required repeat surgery because of an incomplete pyloromyotomy, all were originally operated on by a general surgeon. Analysis of the effect of surgeon volume on outcomes suggested that higher volume resulted in better outcome in both groups. CONCLUSION: Subspecialist pediatric general surgeons achieve superior outcomes for children who undergo Ramstedt pyloromyotomy. PMID- 15121952 TI - Placenta growth factor elevation in the cord blood of premature neonates predicts poor pulmonary outcome. AB - OBJECTIVE: To determine whether an elevated placenta growth factor (PlGF) level in cord blood is associated with increased risk for preterm infants to develop bronchopulmonary dysplasia (BPD). METHODS: Sixty-three preterm infants who were born at 34 weeks' gestation or earlier were enrolled. Two infants who died before 28 days' postnatal age could not be assigned a BPD status and were excluded. PlGF levels in cord blood were measured using enzyme-linked immunosorbent assay. Mann Whitney rank sum test, Spearman correlation coefficients, and multivariable linear or logistic regression analyses were used for statistical analysis. RESULTS: The BPD group had a higher PlGF level, lower gestational age, lower birth weight (BW), higher incidence of endotracheal tube intubation, and longer duration of intubation. The PlGF levels in cord blood correlated negatively with gestational age and BW. However, multivariable logistic regression analyses revealed that only elevated cord blood PlGF levels and BW were associated with BPD after adjusting for all contributing factors. Furthermore, an increased PlGF level in cord blood was significantly correlated with the clinical severity of BPD, as measured by duration of intubation. At 17 mg/dL, the specificity of cord blood PlGF level in predicting BPD was 95%, the sensitivity was 53%, the positive predictive value was 83%, and the negative predictive value was 82%. CONCLUSIONS: Measuring cord blood PlGF level at birth might be a biological marker for predicting the occurrence of BPD and allowing early therapeutic intervention. PMID- 15121953 TI - Initial and follow-up costs by treatment outcome for children with respiratory infections. AB - OBJECTIVE: To estimate the initial and follow-up costs of treatment of respiratory infections among pediatric patients and the relationship between costs and outcomes. METHODS: A total of 3677 episodes of care from 2328 patients who were <17 years of age and had respiratory or ear infections that were treated with an antibiotic initially. The sample was drawn from the Medical Expenditure Panel Surveys for the years 1996 through 1999. Treatment failure was defined as the receipt of a second antibiotic, different from the first, in a 4-week window. We compared follow-up costs by outcome (treatment failure vs success) using a 2 part model of medical costs. We also performed a paired analysis by selecting 2 episodes, one in which the outcome was failure and the other in which the outcome was success, for patients with at least 1 of each type and computing the difference in costs. RESULTS: Follow-up costs for provider visits for episodes for which the patient experienced treatment failure were 216 dollars versus 53 dollars for episodes for which the patient did not experience treatment failure. Follow-up drug costs, including the cost of the second antibiotic, were 75 dollars for children who experienced treatment failure versus 23 dollars, respectively. Cost estimates from the paired analysis were similar, confirming that results are not biased by unobserved time-invariant patient characteristics. CONCLUSION: Children who have respiratory infections and experience treatment failure incur substantially higher costs. PMID- 15121954 TI - Reactions and needs of tristate-area pediatricians after the events of September 11th: implications for children's mental health services. AB - OBJECTIVE: The terrorist attacks of September 11, 2001, caused mass destruction in Lower Manhattan, the Pentagon, and rural Pennsylvania and resulted in the death of >3000 people. Children were prominent among those affected. Given the wide impact of the attacks, we hypothesized that primary care professionals would see the broad population of affected children but would feel ill-prepared to respond to children's mental health needs. METHODS: One year after the September 11th disaster, a hyperlink to a web-based 42-item survey was sent to all New York, Connecticut, and New Jersey American Academy of Pediatrics members with e mail addresses (N = 4330), and a paper version of the survey was sent via postal mail to a random sample of those without e-mail (N = 1320). The survey requested demographic data, personal and practice experience of 9/11, perceived knowledge and skills regarding mental health, and perceived barriers to accessing mental health services for their patients. Both groups were contacted a total of 3 times at 2-week intervals, resulting in 1396 completed surveys from providers who were actively seeing patients. RESULTS: Twenty-nine percent of respondents stated that they were seeing affected patients, and 32.6% reported seeing children who were exposed to at least 1 9/11 event. Sixty-four percent of the respondents identified behavioral problems in directly affected children: 41.6% identified acute stress disorder, and 26.3% identified posttraumatic stress disorder (PTSD). However, a majority of these professionals indicated that they either lacked or were uncertain (50.8% PTSD, 51.7% acute stress disorder) of their skills to identify children with mental health problems and that they were "not" or only "somewhat" knowledgeable (76.8% PTSD) in these areas. The majority agreed that child health professionals should be trained to screen for these 2 disorders. Generalists as compared with specialists were more likely to report seeing patients who were affected by 9/11. Gender, race/ethnicity, and geographic location were associated with reported effects of 9/11 on respondents' practice and perceived skills and knowledge related to the psychological effects of community disasters. CONCLUSIONS: Pediatric practitioners in the tristate area reported that children/families sought care for an array of mental health-related concerns. Generalists in the areas affected and those who identified gaps in knowledge or skills in responding to the psychological effects of community disasters should be targeted for additional education. PMID- 15121955 TI - Unmet need for counseling services by children in New York City after the September 11th attacks on the World Trade Center: implications for pediatricians. AB - OBJECTIVE: The objectives of this study were to describe the prevalence of counseling services, contrasted with the need after the terrorist attacks of September 11, 2001, the types of counseling received, and the predictors of receipt of counseling services. METHODS: A cross-sectional, random-digit-dial survey was conducted in New York City (NYC) of parents (N = 434) of children who were 4 to 17 years of age 4 months after the September 11th terrorist attacks on the World Trade Center. RESULTS: Overall, 10% of NYC children received some type of counseling after the September 11th attacks, according to parental report. Among these, 44% received counseling in schools, 36% received counseling from medical or professional providers, and 20% received counseling from other sources. However, only 27% of the children who had severe/very severe posttraumatic stress reactions (PTSR) after the attacks received counseling services. In a multivariate model, receipt of counseling before the September 11th attacks (odds ratio: 4.44) and having severe/very severe PTSR (odds ratio: 3.59) were the most important predictors of use of counseling services after the September 11th attacks. Minority status and having a parent who experienced the loss of a friend or a relative were also associated with receipt of services. CONCLUSIONS: There was substantial disparity between apparent need (as indicated by severe/very severe PTSR) for and receipt of mental health services for children after the September 11th attacks. There is need for intensified efforts to identify, refer, and treat children in need, especially for children who are not already in a therapeutic relationship. An enhanced role for pediatricians is indicated. PMID- 15121957 TI - Pediatric sleep medicine comes into its own. PMID- 15121956 TI - West Nile virus infection: a pediatric perspective. AB - West Nile virus (WNV) infection recently became a major public health concern in the western hemisphere. This article describes recent information regarding previously unrecognized mechanisms of WNV transmission and reviews clinical manifestations of WNV infection, diagnostic tests, and prevention strategies from a pediatric perspective. WNV is transmitted to humans primarily through the bite of infected mosquitoes, but during the epidemic that spread across North America in 2002, transmission of WNV through blood transfusions and organ transplantation was described for the first time. Individual case reports indicate that WNV can be transmitted also in utero and probably through breast milk. Although most WNV infections are asymptomatic, the virus causes a broad range of manifestations from uncomplicated febrile illness to meningitis, neuropathies, paralysis, and encephalitis. Severe manifestations of WNV infection are far more common in adults than in children, but 105 cases of neuroinvasive WNV disease were reported among children in the United States in 2002. The distribution of the virus in North America continues to spread. WNV infection can be diagnosed by detecting WNV-specific antibody in cerebrospinal fluid or serum, or by detecting the virus or viral nucleic acid in cerebrospinal fluid, blood, or tissues. Cornerstones of prevention include personal protection against mosquitoes, including wearing insect repellent, reducing populations of vector mosquitoes, and screening the blood supply for WNV-contaminated blood donations. PMID- 15121958 TI - The misuse of race in medical diagnosis. PMID- 15121959 TI - Hospital-acquired hyponatremia: why are there still deaths? PMID- 15121960 TI - More than a matter of time. PMID- 15121961 TI - Addressing the Accreditation Council for Graduate Medical Education competencies: an opportunity to impact medical education and patient care. PMID- 15121963 TI - Should we test 4-year-olds? PMID- 15121962 TI - Are we ready and willing to address the mental health needs of children? Implications from September 11th. PMID- 15121964 TI - Intramedullary hemorrhage in a neonate after lumbar puncture resulting in paraplegia: a case report. AB - We present the case of a premature infant with decreased spontaneous movement of the lower extremities. Imaging was demonstrative of a lesion of the conus medullaris. Operatively and with histologic confirmation, the mass was determined to be a blood clot originating from the conus. Retrospectively, the patient had a known lumbar puncture. There were no clotting abnormalities in this patient. At long-term follow-up, the child continues to have lower extremity paresis and incontinence of bowel and bladder. Clinicians should consider the lower termination of the conus medullaris in the infant, especially in the preterm infant. PMID- 15121965 TI - Fathers and pediatricians: enhancing men's roles in the care and development of their children. AB - Research substantiates that fathers' interactions with their children can exert a positive influence on their children's development. This report suggests ways pediatricians can enhance fathers' caregiving involvement by offering specific, culturally sensitive advice and how pediatricians might change their office practices to support and increase fathers' active involvement in their children's care and development. PMID- 15121966 TI - Otitis media with effusion. AB - The clinical practice guideline on otitis media with effusion (OME) provides evidence-based recommendations on diagnosing and managing OME in children. This is an update of the 1994 clinical practice guideline "Otitis Media With Effusion in Young Children," which was developed by the Agency for Healthcare Policy and Research (now the Agency for Healthcare Research and Quality). In contrast to the earlier guideline, which was limited to children 1 to 3 years old with no craniofacial or neurologic abnormalities or sensory deficits, the updated guideline applies to children aged 2 months through 12 years with or without developmental disabilities or underlying conditions that predispose to OME and its sequelae. The American Academy of Pediatrics, American Academy of Family Physicians, and American Academy of Otolaryngology-Head and Neck Surgery selected a subcommittee composed of experts in the fields of primary care, otolaryngology, infectious diseases, epidemiology, hearing, speech and language, and advanced practice nursing to revise the OME guideline. The subcommittee made a strong recommendation that clinicians use pneumatic otoscopy as the primary diagnostic method and distinguish OME from acute otitis media. The subcommittee made recommendations that clinicians should 1) document the laterality, duration of effusion, and presence and severity of associated symptoms at each assessment of the child with OME, 2) distinguish the child with OME who is at risk for speech, language, or learning problems from other children with OME and more promptly evaluate hearing, speech, language, and need for intervention in children at risk, and 3) manage the child with OME who is not at risk with watchful waiting for 3 months from the date of effusion onset (if known) or diagnosis (if onset is unknown). The subcommittee also made recommendations that 4) hearing testing be conducted when OME persists for 3 months or longer or at any time that language delay, learning problems, or a significant hearing loss is suspected in a child with OME, 5) children with persistent OME who are not at risk should be reexamined at 3- to 6-month intervals until the effusion is no longer present, significant hearing loss is identified, or structural abnormalities of the eardrum or middle ear are suspected, and 6) when a child becomes a surgical candidate (tympanostomy tube insertion is the preferred initial procedure). Adenoidectomy should not be performed unless a distinct indication exists (nasal obstruction, chronic adenoiditis); repeat surgery consists of adenoidectomy plus myringotomy with or without tube insertion. Tonsillectomy alone or myringotomy alone should not be used to treat OME. The subcommittee made negative recommendations that 1) population-based screening programs for OME not be performed in healthy, asymptomatic children, and 2) because antihistamines and decongestants are ineffective for OME, they should not be used for treatment; antimicrobials and corticosteroids do not have long-term efficacy and should not be used for routine management. The subcommittee gave as options that 1) tympanometry can be used to confirm the diagnosis of OME and 2) when children with OME are referred by the primary clinician for evaluation by an otolaryngologist, audiologist, or speech-language pathologist, the referring clinician should document the effusion duration and specific reason for referral (evaluation, surgery) and provide additional relevant information such as history of acute otitis media and developmental status of the child. The subcommittee made no recommendations for 1) complementary and alternative medicine as a treatment for OME, based on a lack of scientific evidence documenting efficacy, or 2) allergy management as a treatment for OME, based on insufficient evidence of therapeutic efficacy or a causal relationship between allergy and OME. Last, the panel compiled a list of research needs based on limitations of the evidence reviewed. The purpose of this guideline is to inform clinicians of evidence-based methods to identify, monitor, and manage OME in children aged 2 months through 12 years. The guideline may not apply to children more than 12 years old, because OME is uncommon and the natural history is likely to differ from younger children who experience rapid developmental change. The target population includes children with or without developmental disabilities or underlying conditions that predispose to OME and its sequelae. The guideline is intended for use by providers of health care to children, including primary care and specialist physicians, nurses and nurse practitioners, physician assistants, audiologists, speech-language pathologists, and child-development specialists. The guideline is applicable to any setting in which children with OME would be identified, monitored, or managed. This guideline is not intended as a sole source of guidance in evaluating children with OME. Rather, it is designed to assist primary care and other clinicians by providing an evidence-based framework for decision-making strategies. It is not intended to replace clinical judgment or establish a protocol for all children with this condition and may not provide the only appropriate approach to diagnosing and managing this problem. PMID- 15121967 TI - Admission and discharge guidelines for the pediatric patient requiring intermediate care. AB - During the past 3 decades, the specialty of pediatric critical care medicine has grown rapidly, leading to a number of pediatric intensive care units opening across the country. Many patients who are admitted to the hospital require a higher level of care than routine inpatient general pediatric care, yet not to the degree of intensity of pediatric critical care; therefore, an intermediate care level has been developed in institutions providing multidisciplinary subspecialty pediatric care. These patients may require frequent monitoring of vital signs and nursing interventions, but usually they do not require invasive monitoring. The admission of the pediatric intermediate care patient is guided by physiologic parameters depending on the respective organ system involved relative to an institution's resources and capacity to care for a patient in a general care environment. This report provides admission and discharge guidelines for intermediate pediatric care. Intermediate care promotes greater flexibility in patient triage and provides a cost-effective alternative to admission to a pediatric intensive care unit. This level of care may enhance the efficiency of care and make health care more affordable for patients receiving intermediate care. PMID- 15121968 TI - Hospital stay for healthy term newborns. AB - Decisions regarding the length of hospital stays for newborns and their mothers became driven by financial reimbursement from third-party payers in the 1990s. The Newborns' and Mothers' Health Protection Act of 1996 and a report from the Secretary's Advisory Committee on Infant Mortality acknowledge the importance of physician assessment in determining the timing of each newborn's discharge. The pediatrician's primary role is to ensure the health and well-being of the newborn in the context of the family. It is within this context that this revised statement addresses the short hospital stay (<48 hours after birth) for healthy term newborns. PMID- 15121969 TI - Application of the resource-based relative value scale system to pediatrics. AB - In today's rapidly changing health care environment, it is crucial to understand the genesis and principles behind the Medicare Resource-Based Relative Value Scale (RBRVS) physician fee schedule. Many third-party payers, including state Medicaid programs, BlueCross BlueShield, and managed care organizations, use variations of the Medicare RBRVS to determine physician reimbursement and capitation rates. Because the RBRVS fee schedule was created originally for Medicare only, pediatric-specific Current Procedural Terminology (CPT) codes and pediatric practice expense calculations were not included. The American Academy of Pediatrics supports the use of CPT codes and the RBRVS physician fee schedule and continues to work to rectify certain inequities of the RBRVS system as they pertain to pediatrics. PMID- 15121970 TI - Recommendations for influenza immunization of children. AB - Epidemiologic studies indicate that children of all ages with certain chronic conditions and otherwise healthy children younger than 24 months of age are hospitalized for influenza infection and its complications at high rates similar to those experienced by the elderly. Annual influenza immunization is recommended for all children with high-risk conditions who are 6 months of age and older. Young, healthy children are at high risk of hospitalization for influenza infection; therefore, the American Academy of Pediatrics recommends influenza immunization for healthy children 6 through 24 months of age, for household contacts and out-of-home caregivers of all children younger than 24 months of age, and for health care professionals. To protect these children more fully against the complications of influenza, increased efforts are needed to identify all high-risk children and inform their parents when annual immunization is due. The purposes of this statement are to update recommendations for routine use of influenza vaccine in children and to review the indications for use of trivalent inactivated influenza vaccine and live-attenuated influenza vaccine. PMID- 15121971 TI - Recommended childhood and adolescent immunization schedule--United States, July December 2004. PMID- 15121972 TI - Diagnosis and management of acute otitis media. AB - This evidence-based clinical practice guideline provides recommendations to primary care clinicians for the management of children from 2 months through 12 years of age with uncomplicated acute otitis media (AOM). The American Academy of Pediatrics and American Academy of Family Physicians convened a committee composed of primary care physicians and experts in the fields of otolaryngology, epidemiology, and infectious disease. The subcommittee partnered with the Agency for Healthcare Research and Quality and the Southern California Evidence-Based Practice Center to develop a comprehensive review of the evidence-based literature related to AOM. The resulting evidence report and other sources of data were used to formulate the practice guideline recommendations. The focus of this practice guideline is the appropriate diagnosis and initial treatment of a child presenting with AOM. The guideline provides a specific definition of AOM. It addresses pain management, initial observation versus antibacterial treatment, appropriate choices of antibacterials, and preventive measures. Decisions were made based on a systematic grading of the quality of evidence and strength of recommendations, as well as expert consensus when definitive data were not available. The practice guideline underwent comprehensive peer review before formal approval by the partnering organizations. This clinical practice guideline is not intended as a sole source of guidance in the management of children with AOM. Rather, it is intended to assist primary care clinicians by providing a framework for clinical decision-making. It is not intended to replace clinical judgment or establish a protocol for all children with this condition. These recommendations may not provide the only appropriate approach to the management of this problem. PMID- 15121973 TI - Did "controversies concerning vitamin K and the newborn" cover all the controversies? PMID- 15121974 TI - Acute psychosis associated with therapeutic use of dextroamphetamine. PMID- 15121975 TI - Bilateral hearing impairment in Dutch neonatal intensive care unit infants with unilateral failure on hearing screening. PMID- 15121976 TI - Paintball injuries in children: the cases managed out of hospitals. PMID- 15121977 TI - Solution to a crushing dosage problem? PMID- 15121978 TI - Car-seat test. PMID- 15121979 TI - Racial/ethnic variation in parent expectations for antibiotics: implications for public health campaigns. AB - CONTEXT: Widespread overuse and inappropriate use of antibiotics are a major public health concern. Little is known about racial/ethnic differences in parents seeking antibiotics for their children's upper respiratory illnesses. OBJECTIVE: To examine racial/ethnic differences in parent expectations about the need for antibiotics and physician perceptions of those expectations. DESIGN: We conducted a nested, cross-sectional survey of parents who were coming to see their child's pediatrician because of cold symptoms between October 2000 and June 2001. Parents completed a previsit survey that collected information on demographics, their child's illness, and a 15-item previsit expectations inventory that included an item asking how necessary it was for the physician to prescribe antibiotics. Physicians completed a postvisit survey that collected information on diagnosis, treatment, and whether the physician perceived the parent expected an antibiotic. The encounter was the unit of analysis. Multivariate logistic regression analyses were performed to evaluate predictors of dichotomized parental expectations for antibiotics, dichotomized physician perceptions of those expectations, diagnostic patterns, and antibiotic-prescribing patterns. SETTING: Twenty-seven community pediatric practices in the Los Angeles, Calif, metropolitan area. PARTICIPANTS: A volunteer sample of 38 pediatricians (participation rate: 64%) and a consecutive sample of 543 parents (participation rate: 83%; approximately 15 participating for each enrolled pediatrician) seeking care for their children's respiratory illnesses. Pediatricians were eligible to participate if they worked in a community-based managed care practice in the Los Angeles area. Parents were eligible to participate if they could speak and read English and presented to participating pediatricians with a child 6 months to 10 years old who had cold symptoms but had not received antibiotics within 2 weeks. MAIN OUTCOME MEASURES: Parental beliefs about the necessity of antibiotics for their child's illness, physician perceptions of parental expectations for antibiotics, bacterial diagnosis rates, and antibiotic-prescribing rates. RESULTS: Forty-three percent of parents believed that antibiotics were definitely necessary, and 27% believed that they were probably necessary for their child's illness. Latino and Asian parents were both 17% more likely to report that antibiotics were either definitely or probably necessary than non-Hispanic white parents. Physicians correctly perceived that Asian parents expected antibiotics more often than non Hispanic white parents but underestimated the greater expectations of Latino parents for antibiotics. Physicians also correctly perceived that parents of children with ear pain or who were very worried about their child's condition were significantly more likely to expect antibiotics. Physicians were 7% more likely to make a bacterial diagnosis and 21% more likely to prescribe antibiotics when they perceived that antibiotics were expected. CONCLUSIONS: Parent expectations for antibiotics remain high in Los Angeles County. With time, traditional public health messages related to antibiotic use may decrease expectations among non-Hispanic white parents. However, both public health campaigns and physician educational efforts may need to be designed differently to reach other racial/ethnic groups effectively. Despite public health campaigns to reduce antibiotic overprescribing in the pediatric outpatient setting, physicians continue to respond to parental pressure to prescribe them. To effectively intervene to decrease rates of inappropriate antibiotic prescribing further, physicians need culturally appropriate tools to better communicate and negotiate with parents when feeling pressured to prescribe antibiotics. PMID- 15121980 TI - Improved access and quality of care after enrollment in the New York State Children's Health Insurance Program (SCHIP). AB - BACKGROUND: Although many studies have noted that uninsured children have poorer access and quality of health care than do insured children, few studies have been able to demonstrate the direct benefits of providing health insurance to previously uninsured children. The State Children's Health Insurance Program (SCHIP), enacted as Title XXI of the Social Security Act, was intended to improve insurance coverage and access to health care for low-income, uninsured children. With limited state and federal resources for health care, continued funding of SCHIP requires demonstration of success of the program. As yet, little is known about the effectiveness of SCHIP on improving access and quality of care to enrollees. OBJECTIVES: To measure the impact of the New York State (NYS) SCHIP on access, utilization, and quality of health services for enrolled children. DESIGN SETTING: NYS, stratified into 4 regions. The NYS SCHIP is modeled on commercial insurance (32 managed care plans) and at the time of the study had 18% of SCHIP enrollees nationwide. STUDY DESIGN: For the study group, the design used pre/poststudy telephone interviews of parents of children enrolling in the NYS SCHIP, with baseline interviews soon after enrollment and follow-up interviews 1 year after enrollment. Baseline interviews reflected the child's experience during the 1-year period before enrollment in SCHIP. The follow-up interviews reflected the 1-year period after enrollment in SCHIP. For the comparison group, the design used baseline interviews of a comparison group enrolled 1 year after the study group to test for secular trends; these interviews reflected the 1-year period before enrollment in SCHIP. SUBJECTS: Children (n = 2644) 0 to 18 years of age who enrolled in the NYS SCHIP for the first time (November 2000 to March 2001), stratified by age (0-5, 6-11, and 12-18 years), race/ethnicity (white non Hispanic, black non-Hispanic, and Hispanic; others excluded), and region of NYS. The comparison group consisted of 400 children. Telephone interviews were conducted in English or Spanish throughout the day and evening, 7 days per week, to obtain measures. MAIN OUTCOME MEASURES: Demographic and health measures (child and family characteristics, health status, presence of a special health care need, and prior health insurance), access (usual source of care [USC] and unmet needs for health care), utilization (visits for specific health services), and quality (continuity with USC and measures of primary care interactions). Analyses included bivariate tests, comparing the pre-SCHIP period to the 1-year period after enrollment in SCHIP. Multivariate models were computed to generate standardized populations comprised of key characteristics of the sample to test for differences in measures (after SCHIP versus before SCHIP), controlling for demographic characteristics. RESULTS: Of the 2644 study-group children who completed the initial interview, 2290 (87%) completed the follow-up interview. Key measures for the pre-SCHIP period and short-term "postenrollment" measures for the study group were not statistically different from measures for the comparison group, suggesting no major secular trends. Participants were non Hispanic white (25%), non-Hispanic black (31%), and Hispanic (45%). Fifty-one percent of the parents were single, and 61% had a high school education or less; 81% of families had income <160% of the federal poverty level. Sixty-two percent of the children were uninsured > or = 12 months before the NYS SCHIP; of those insured, 43% previously had Medicaid. The proportion of children who had a USC increased after enrollment in the NYS SCHIP (86% to 97%). Two measures of accessibility (difficulty getting a medical person by telephone and difficulty getting an appointment) improved after enrollment in SCHIP. The proportion of children with any unmet health care needs decreased (31% to 19%). Specific types of unmet need also were reduced after enrollment; for example, among SCHIP enrollees who had a need for specific type of care, unmet needs wds were significantly lower postenrollment versus pre-SCHIP for specialty care (-15.5% in unmet need), acute care (-10.1%), preventive care (-9.6%), dental care (-13.0%%), and vision care (-13.2%). Emergency and total ambulatory visits did not change, but the proportion of children with a preventive care visit increased (74% to 82%). The proportion of children who used their USC for most or all visits increased (47% to 89%), demonstrating increased continuity of care. Several indicators of health care quality improved, including an overall rating of quality, the 4 indicators of physician-patient interaction used by the Consumer Assessment of Health Plans Survey, and a measure of parental worry about their child's health. Improvements were noted among major subgroups of children, with the greatest improvements for those with the lowest baseline levels. For example, at baseline, a lower percentage of children living at <160% of the federal poverty level had a presence of a USC or continuity with their USC than children living in families at >160% of the federal poverty level, and these poorer children experienced the greatest gains in having a USC or having continuity with their USC after enrollment in SCHIP. CONCLUSIONS: Enrollment in the NYS SCHIP was associated with 1) improved access, continuity, and quality of care and 2) a change in the pattern of health care, with a greater proportion of care taking place within the usual source of primary care. PMID- 15121982 TI - Efficacy of automated continuous positive airway pressure in children with sleep related breathing disorders in an attended setting. AB - INTRODUCTION: The purpose of this study was to evaluate the safety and efficacy of automated continuous positive airway pressure (Auto-CPAP) in children. Sleep related breathing disorders (SRBDs) include the clinical spectrum of symptomatic chronic snoring, upper airway resistance syndrome, and obstructive sleep apnea. This spectrum occurs in adults and children. Less data are available for children despite recognition of the condition's prevalence. CPAP has been an established treatment for adults and children. Treatment with Auto-CPAP has been available for adults but has not been reported previously in children. METHODS: A group of 14 children (8 months to 12 years old) was evaluated prospectively with baseline polysomnographic study and CPAP titration performed with Auto-CPAP under sleep technologist supervision. RESULTS: The results demonstrated that Auto-CPAP is sensitive and effective for children with obstructive sleep apnea in an attended setting. There was 1 subject who did not seem to tolerate Auto-CPAP, but when she was switched to conventional CPAP, she did not tolerate that either. In this subject, the mask never fit well. She was excluded from the analysis. All other patients had a decrease in the number of abnormal breathing events during sleep. The respiratory disturbance index decreased from a mean of 12.6 (SD: 12.4) to 2.6 (SD: 2.7) events per hour. The lowest oxygen saturation improved from a mean of 86% (SD: 10.8) to 93.6% (SD: 3.9). CONCLUSIONS: We conclude that Auto-CPAP is safe and effective in an attended environment. Auto-CPAP did not eliminate all the abnormal respiratory events. In subjects 1 and 14, the final respiratory index improved but remained >5 events per hour (5.9 and 7.7, respectively). We suspect that this was because of problems with the masks leaking, which illustrates the importance of follow-up and possible need for retitration in some patients. Proper mask fit is essential for successful treatment. Additional work is needed to evaluate its utility in the home setting. This study was designed to evaluate Auto-CPAP titration in an attended environment. It did not indicate information about the effectiveness in an unattended or home setting. We demonstrate that Auto-CPAP is able to detect abnormal breathing events during sleep in children and may provide the necessary pressure to correct these events. Auto-CPAP can be used safely for pressure titration in an attended setting. Auto CPAP devices from different manufactures are commercially available for adults. These different devices may have different algorithms and sensitivities to detect abnormal breathing episodes. This study was performed with only 1 specific model of Auto-CPAP. Our results should not be extrapolated to other Auto-CPAP devices without empirical confirmation of the devices' ability to detect and correct events in children. Auto-CPAP can be an alternative treatment for SRBDS in the pediatric population. These results allow for speculation of possible applications for Auto-CPAP in children. A potential advantage of Auto-CPAP includes permitting the initiation of treatment while awaiting a standard CPAP titration. The variable pressure response of Auto-CPAP allows for treatment under different situations such as upper airway infections, different sleeping positions, and changes in weight. As the child grows, the amount of positive pressure needed to maintain airway patency may change. Auto-CPAP may be able to adjust to these changing pressure requirements. Auto-CPAP does not eliminate the need for periodic office visits and evaluations of the clinical course. PMID- 15121981 TI - Mothers' and clinicians' perspectives on breastfeeding counseling during routine preventive visits. AB - BACKGROUND: Recent national statistics indicate that, despite increases in the proportion of mothers who initiate breastfeeding, the proportion that continue to breastfeed their infants through 6 months of age remains below the Healthy People 2010 goal of 50%. National professional organizations recommend that clinicians routinely counsel mothers about the benefits of breastfeeding. Little is known, however, about the counseling provided during these visits and how mothers and their clinicians perceive breastfeeding counseling. OBJECTIVES: We sought to describe mothers' and clinicians' perspectives on breastfeeding counseling during routine preventive visits and identify potential gaps in communication about breastfeeding and management practices. METHODS: We conducted a prospective cohort study of low-risk mother-newborn pairs and their clinicians in a large multispecialty group practice. The participating mothers completed telephone interviews at 4 and 12 weeks postpartum, and their data were linked with their obstetric and pediatric clinicians' responses to a cross-sectional mailed survey conducted during the same time period. Overall, response rates were 63% for mothers (n = 429) and 82% for clinicians (obstetric clinicians: n = 54; pediatric clinicians: n = 67). RESULTS: Of the 429 low-risk mother-newborn pairs in the study, 61% were white, 16% were black, 10% were Hispanic, and 8% were Asian, with a mean (SD) age of 32.7 (5.1) years. At 4 weeks postpartum, 319 mothers (74%) were either exclusively or mixed breastfeeding. According to the interviews, few mothers discussed breastfeeding duration with their obstetric clinicians during their prenatal visits (15%) or with their pediatric clinicians during their infants' 2-week preventive visit (24%). Among 164 mothers whose obstetric providers said they usually or always discuss breastfeeding duration during prenatal visits, only 26 (16%) of the mothers reported that the topic was discussed with them (22% agreement; kappa = -.004). Among those mothers whose pediatric clinicians said they usually or always discuss breastfeeding duration during the 2-week preventive visit, only 25% of the mothers reported that the topic was discussed (32% agreement; kappa =.05). Many of the mothers had either returned to work by 12 weeks (29%) or planned to return to work within the next few months (43%). Although nearly all the obstetric (91%) and pediatric (97%) clinicians reported that they usually or always discuss whether a mother plans to continue breastfeeding after returning to work, only approximately half (55%) of the mothers seen by the clinicians reported that the topic was discussed. Overall, few mothers reported discussing with their clinicians specific ways to continue breastfeeding after returning to work. CONCLUSION: Mothers' reports of breastfeeding advice given during routine preventive visits identified several areas in which unintentional communication gaps may occur, including specifics about breastfeeding duration and methods of breastfeeding after returning to work. Developing approaches to enhance communication with mothers during routine preventive visits could improve the support of breastfeeding. PMID- 15121983 TI - The temporal relationship between the onset of type 1 diabetes and celiac disease: a study based on immunoglobulin a antitransglutaminase screening. AB - OBJECTIVE: The association of celiac disease (CD) and type 1 diabetes is now clearly documented. Immunoglobulin A (IgA) antitransglutaminase antibodies were measured to determine the prevalence of celiac disease in a diabetic population of children and to determine the temporal relationship between type 1 diabetes onset and CD. METHODS: We measured IgA antitransglutaminase antibodies using human recombinant antigen in parallel with classical markers (IgA and IgG antigliadin, IgA antiendomysium) in 284 children with diabetes. RESULTS: In the population studied, the prevalence of CD was 3.9% (11 of 284). Two cases of CD were diagnosed before the onset of diabetes, and in 8 patients, the diagnoses of CD and diabetes were concomitant, suggesting that CD was present before the onset of diabetes. In 1 case, a girl who presented with thyroiditis, serology for CD became positive after diabetes had been diagnosed. CONCLUSION: An excellent correlation was observed between IgA antiendomysium and IgA antitransglutaminase antibodies. We therefore propose using IgA antitransglutaminase as a screening test for practical reasons. Furthermore, IgA antitransglutaminase levels and mucosa abnormalities were closely correlated. The presence of antitransglutaminase antibodies should alert pediatricians to the atypical forms of CD. This study indicates that CD is most often present before the onset of diabetes. PMID- 15121984 TI - Living with classical galactosemia: health-related quality of life consequences. AB - OBJECTIVE: Classical galactosemia (McKusick 230400) is an autosomal recessive disorder of galactose metabolism caused by a deficiency of galactose-1-phosphate uridyltransferase (EC 2.7.712). Treatment, consisting of a severe restriction of dietary galactose, is life saving, but most patients develop abnormalities despite this diet. The aim of this study was to study the influence of galactosemia on the patients' health-related quality of life (HRQoL), on educational levels, and on the specific galactosemia-related concerns of these families. METHODS: Age-specific HRQoL questionnaires, a classical galactosemia specific questionnaire designed by the authors, and a list of questions regarding educational attainment were handed out or sent to all 75 members of the Dutch Galactosemia Society and their families. RESULTS: Sixty-three (84%) patients with classical galactosemia from 58 families returned the questionnaire. Concerning HRQoL, significant differences between patients aged 1 to 5 and healthy children were found on the domains of abdominal complaints and communication. Patients aged 8 to 15 years differed from their healthy peers on the domain of cognitive function. Mothers of patients aged 6 to 15 reported a significantly lower HRQoL on the domains of motor and cognitive function. Patients 16 years and older had significant lower scores on the domains of cognitive and social function. The percentage of patients who attend special schools is significantly higher than in the general population, and the educational attainment is significantly lower in patients with classical galactosemia. CONCLUSIONS: This is the first study to describe the HRQoL of patients with classical galactosemia using well-developed and validated instruments in different age groups. The results of the present study indicate that having galactosemia negatively influences the HRQoL. Early and regular evaluation and support of possible cognitive problems should be a major part of the protocol for the follow-up of patients with classical galactosemia. PMID- 15121986 TI - Breastfeeding and the risk of postneonatal death in the United States. AB - OBJECTIVE: Breastfed infants in the United States have lower rates of morbidity, especially from infectious disease, but there are few contemporary studies in the developed world of the effect of breastfeeding on postneonatal mortality. We evaluated the effect of breastfeeding on postneonatal mortality in United States using 1988 National Maternal and Infant Health Survey (NMIHS) data. METHODS: Nationally representative samples of 1204 infants who died between 28 days and 1 year from causes other than congenital anomaly or malignant tumor (cases of postneonatal death) and 7740 children who were still alive at 1 year (controls) were included. We calculated overall and cause-specific odds ratios for ever/never breastfeeding among all children, conducted race and birth weight specific analyses, and looked for duration-response effects. RESULTS: Overall, children who were ever breastfed had 0.79 (95% confidence interval [CI]: 0.67 0.93) times the risk of never breastfed children for dying in the postneonatal period. Longer breastfeeding was associated with lower risk. Odds ratios by cause of death varied from 0.59 (95% CI: 0.38-0.94) for injuries to 0.84 (95% CI: 0.67 1.05) for sudden infant death syndrome. CONCLUSIONS: Breastfeeding is associated with a reduction in risk for postneonatal death. This large data set allowed robust estimates and control of confounding, but the effects of breast milk and breastfeeding cannot be separated completely from other characteristics of the mother and child. Assuming causality, however, promoting breastfeeding has the potential to save or delay approximately 720 postneonatal deaths in the United States each year. PMID- 15121985 TI - Exposure to di-(2-ethylhexyl) phthalate among premature neonates in a neonatal intensive care unit. AB - OBJECTIVE: Premature neonates who spend time in a neonatal intensive care unit may be at increased risk of adverse health effects from exposure to di-(2 ethylhexyl) phthalate (DEHP) because of their increased risk of high exposure, their small body size, and their physical condition. DEHP, a reproductive toxicant in animals, is a major component in polyvinyl chloride (PVC) plastics, which are frequently used in medical tubing and blood storage bags. DEHP is not covalently bound to PVC, and it may be easily released from the PVC medical devices. The objective of this study was to determine whether premature infants who undergo medical procedures, such as blood transfusions, intravenous therapy, enteral and parenteral nutrition support, and dialysis, are at increased risk of exposure to DEHP than the general population. Because of their smaller size, children and especially premature and small infants may receive a larger dose of DEHP on a milligram per kilogram basis than adults when the same-size medical device is used for all ages. METHODS: Premature neonates who seemed to have the potential to be on intravenous infusion for >2 weeks and were expected to survive were eligible for enrollment in the study. We assessed exposure to DEHP in 6 premature newborns by measuring in 41 urine samples the levels of 3 DEHP metabolites: mono-(2-ethylhexyl) phthalate (mEHP), mono-(2-ethyl-5-hydroxyhexyl) phthalate (mEHHP), and mono-(2-ethyl-5-oxohexyl) phthalate (mEOHP). RESULTS: mEHHP and mEOHP were detected in all 41 urine samples, and mEHP was detected in 33. Because only 33 of the samples had detectable amounts for all 3 metabolites, statistical analyses were limited to those 33. The levels of all 3 DEHP metabolites varied widely, and the urinary mean and median concentrations of mEOHP and mEHHP were 1 order of magnitude higher than those for mEHP. Furthermore, the geometric mean urinary concentrations of mEOHP (1617 ng/mL), mEHHP (2003 ng/mL), and mEHP (100 ng/mL) in these 6 premature infants who underwent intensive therapeutic interventions were found to be severalfold higher than in the US general population (for mEHP, geometric mean in those 6 years and older was 3.43 ng/mL). CONCLUSIONS: This study provides the first quantitative evidence confirming that newborns who undergo intensive therapeutic medical interventions are exposed to higher concentrations of DEHP than the general population. Although the overall benefits of medical procedures using PVC devices outweigh the risks associated with exposure to DEHP, more research is needed to determine whether infants and children who undergo intensive therapeutic interventions using DEHP-containing devices are at higher risk for altered health outcomes than infants and children who undergo similar treatments but are not potentially exposed to DEHP. PMID- 15121987 TI - Failure to thrive and cognitive development in toddlers with infantile anorexia. AB - OBJECTIVE: The goal of this study was to examine the relative contributions of growth deficiency and psychosocial factors to cognitive development in toddlers with infantile anorexia. METHODS: Eighty-eight toddlers, ranging in age from 12 to 33 months, were enrolled in this study. Toddlers were evaluated by 2 child psychiatrists and placed into 1 of 3 groups: infantile anorexia, picky eater, and healthy eater. All 3 groups were matched for age, race, gender, and socioeconomic status (SES). Toddlers underwent nutritional evaluations and cognitive assessments with the Bayley Scales of Infant Development. Toddlers and their mothers were also videotaped during feeding and play interactions, which later were rated independently by 2 observers. RESULTS: On average, toddlers with infantile anorexia performed within the normal range of cognitive development. However, the Mental Developmental Index (MDI) scores of the healthy eater group (MDI = 110) were significantly higher than those of the infantile anorexia (MDI = 99) and picky eater (MDI = 96) groups. Within the infantile anorexia group, correlations between MDI scores and the toddlers' percentage of ideal body weight approached statistical significance (r =.32). Across all groups, the toddlers' MDI scores were associated with the quality of mother-child interactions, SES level, and maternal education level. Collectively, these variables explained 22% of the variance in MDI scores. CONCLUSIONS: This study demonstrated that psychosocial factors, such as mother-toddler interactions, maternal education level, and SES level, are related to the cognitive development of toddlers with feeding problems and explain more unique variance in MDI scores than nutritional status. PMID- 15121988 TI - Long-term intravenous treatment of Pompe disease with recombinant human alpha glucosidase from milk. AB - OBJECTIVE: Recent reports warn that the worldwide cell culture capacity is insufficient to fulfill the increasing demand for human protein drugs. Production in milk of transgenic animals is an attractive alternative. Kilogram quantities of product per year can be obtained at relatively low costs, even in small animals such as rabbits. We tested the long-term safety and efficacy of recombinant human -glucosidase (rhAGLU) from rabbit milk for the treatment of the lysosomal storage disorder Pompe disease. The disease occurs with an estimated frequency of 1 in 40,000 and is designated as orphan disease. The classic infantile form leads to death at a median age of 6 to 8 months and is diagnosed by absence of alpha-glucosidase activity and presence of fully deleterious mutations in the alpha-glucosidase gene. Cardiac hypertrophy is characteristically present. Loss of muscle strength prevents infants from achieving developmental milestones such as sitting, standing, and walking. Milder forms of the disease are associated with less severe mutations and partial deficiency of alpha-glucosidase. METHODS: In the beginning of 1999, 4 critically ill patients with infantile Pompe disease (2.5-8 months of age) were enrolled in a single-center open-label study and treated intravenously with rhAGLU in a dose of 15 to 40 mg/kg/week. RESULTS: Genotypes of patients were consistent with the most severe form of Pompe disease. Additional molecular analysis failed to detect processed forms of alpha-glucosidase (95, 76, and 70 kDa) in 3 of the 4 patients and revealed only a trace amount of the 95-kDa biosynthetic intermediate form in the fourth (patient 1). With the more sensitive detection method, 35S-methionine incorporation, we could detect low-level synthesis of -glucosidase in 3 of the 4 patients (patients 1, 2, and 4) with some posttranslation modification from 110 kDa to 95 kDa in 1 of them (patient 1). One patient (patient 3) remained totally deficient with both detection methods (negative for cross-reactive immunologic material [CRIM negative]). The alpha-glucosidase activity in skeletal muscle and fibroblasts of all 4 patients was below the lower limit of detection (<2% of normal). The rhAGLU was tolerated well by the patients during >3 years of treatment. Anti-rhAGLU immunoglobulin G titers initially increased during the first 20 to 48 weeks of therapy but declined thereafter. There was no consistent difference in antibody formation comparing CRIM-negative with CRIM-positive patients. Muscle alpha-glucosidase activity increased from <2% to 10% to 20% of normal in all patients during the first 12 weeks of treatment with 15 to 20 mg/kg/week. For optimizing the effect, the dose was increased to 40 mg/kg/week. This resulted, 12 weeks later, in normal alpha-glucosidase activity levels, which were maintained until the last measurement in week 72. Importantly, all 4 patients, including the patient without any endogenous alpha-glucosidase (CRIM negative), revealed mature 76- and 70-kDa forms of -glucosidase on Western blot. Conversion of the 110-kDa precursor from milk to mature 76/70-kDa alpha glucosidase provides evidence that the enzyme is targeted to lysosomes, where this proteolytic processing occurs. At baseline, patients had severe glycogen storage in the quadriceps muscle as revealed by strong periodic acid-Schiff- positive staining and lacework patterns in hematoxylin and eosin--stained tissue sections. The muscle pathology correlated at each time point with severity of signs. Periodic acid-Schiff intensity diminished and number of vacuoles increased during the first 12 weeks of treatment. Twelve weeks after dose elevation, we observed signs of muscle regeneration in 3 of the 4 patients. Obvious improvement of muscular architecture was seen only in the patient who learned to walk. Clinical effects were significant. All patients survived beyond the age of 4 years, whereas untreated patients succumb at a median age of 6 to 8 months. The characteristic cardiac hypertrophy present at start of treatment diminished significantly. The left ventricular mass index decreased from 171 to 599 g/m2 (upper limit of normal 86.6 g/m2 for infants from 0 to 1 year) to 70 to 160 g/m2 during 84 weeks of treatment. In addition, we found a significant change of slope for the diastolic thickness of the left ventricular posterior wall against time at t = 0 for each separate patient. Remarkably, the younger patients (patients 1 and 3) showed no significant respiratory problems during the first 2 years of life. One of the younger patients recovered from a life-threatening bronchiolitis at the age of 1 year without sequelae, despite borderline oxygen saturations at inclusion. At the age of 2, however, she became ventilator dependent after surgical removal of an infected Port-A-Cath. She died at the age of 4 years and 3 months suddenly after a short period of intractable fever of >42 degrees C, unstable blood pressure, and coma. The respiratory course of patient 1 remained uneventful. The 2 older patients, who both were hypercapnic (partial pressure of carbon dioxide: 10.6 and 9.8 kPa; normal range: 4.5-6.8 kPa) at start of treatment, became ventilator dependent before the first infusion (patient 2) and after 10 weeks of therapy (patient 4). Patient 4 was gradually weaned from the ventilator after 1 year of high-dose treatment and was eventually completely ventilator-free for 5 days, but this situation could not be maintained. Currently, both patients are completely ventilator dependent. The most remarkable progress in motor function was seen in the younger patients (patients 1 and 3). They achieved motor milestones that are unmet in infantile Pompe disease. Patient 1 learned to crawl (12 months), walk (16 months), squat (18 months), and climb stairs (22 months), and patient 3 learned to sit unsupported. The Alberta Infant Motor Scale score for patients 2, 3, and 4 remained far below p5. Patient 1 followed the p5 of normal. CONCLUSION: Our study shows that a safe and effective medicine can be produced in the milk of mammals and encourages additional development of enzyme replacement therapy for the several forms of Pompe disease. Restoration of skeletal muscle function and prevention of pulmonary insufficiency require dosing in the range of 20 to 40 mg/kg/week. The effect depends on residual muscle function at the start of treatment. Early start of treatment is required. PMID- 15121989 TI - Recent trends in child restraint practices in the United States. AB - OBJECTIVE: To assess the success of recent outreach activities to promote appropriate child restraint in motor vehicles by examining trends in restraint types used by children under age 9 in 3 large regions of the United States. METHODS: Cross-sectional study was conducted of children who were under age 9 and in crashes of insured vehicles in 15 states, with data collected via insurance claims records and a telephone survey. A probability sample of 8730 crashes involving 10,195 children, representing 128 291 crashes involving 149,820 children, was collected between December 1, 1998, and November 30, 2002. Parent report was used to determine restraint type used in the crash. Logistic regression models were used to analyze the secular trend of restraint type use. RESULTS: Overall, for children under age 9, seat belt use decreased significantly from 49% to 36% between 1998 and 2002; for 7- and 8-year-olds, from 97% to 92%; and for 3- to 6-year-olds, from 63% to 34%. Concurrently, gains were achieved in overall child restraint use from 49% to 63%, for 7- and 8-year-olds, from 2% to 5%; and for 3- to 6-year-olds, from 35% to 65%. Child restraint use remains stable for children from birth to 2 years of age (from 97% in 1998 to 98% in 2002). Both the use of child safety seat and belt positioning booster seat increased significantly, whereas shield booster seat use decreased significantly. CONCLUSION: Although considerable achievements have been realized over a short period of time, substantial inappropriate restraint still remains: 62% of children aged 4 to 8 remain inappropriately restrained in adult seat belts. Parents hear safety messages when they are relevant to their children. As a result, sustained efforts about appropriate restraint must continue to maintain and improve the gains achieved in appropriate child restraint use. The additional benefits realized by recent changes in child restraint laws remain to be evaluated. PMID- 15121990 TI - Prepregnant overweight and obesity diminish the prolactin response to suckling in the first week postpartum. AB - OBJECTIVE: The population subgroups with the highest proportion of overweight and obese women often are characterized by the lowest rates of initiation and shortest durations of breastfeeding. We previously documented that these 2 population-level trends may be related. In a population of white women who lived in a rural area, we observed that prepregnant overweight and obesity were associated with failure to initiate and also to sustain lactation. The means by which being overweight or obese negatively affect lactational performance is unknown and likely to be multifactorial in origin, including the simple mechanical difficulties of latching on and proper positioning of the infant. In addition, we have shown that prepregnant body mass index (BMI) is negatively associated with the timing of lactogenesis II, the onset of copious milk secretion. Although the effects of obesity on the prolactin response to infant suckling have never been studied, we postulated that maternal obesity could compromise this important response. We proposed that this might occur because obesity alters the 24-hour spontaneous release of prolactin and also because prolactin secretion is blunted in response to various stimuli among obese subjects. The fall in progesterone concentration that occurs immediately postpartum is the trigger for the onset of copious milk secretion, but maintenance of prolactin and cortisol concentrations is necessary for this trigger to be effective. Adipose tissue concentrates progesterone. We proposed that this additional source of progesterone would lead to consistently higher progesterone concentrations among obese compared with normal-weight women. This, in turn, would lead to a delay in reaching the appropriate concentration to trigger the onset of lactogenesis II. We tested the hypotheses that a reduced prolactin response to suckling and higher-than-normal progesterone concentration in the first week after delivery might be among the means by which maternal overweight could compromise early lactation. METHODS: We enrolled 40 mothers of term infants from the same population that we studied previously. We measured serum prolactin and progesterone concentrations by radioimmunoassay before and 30 minutes after the beginning of a suckling episode at 48 hours and 7 days after delivery. We used path analysis to develop a parsimonious multivariate prediction of the prolactin response to suckling at 48 hours and 7 days postpartum. RESULTS: As expected, prolactin values decreased from 48 hours to 7 days postpartum. Women who were overweight or obese (using the Institute of Medicine's cutoff for women of a BMI >26 kg/m2) before conception had a lower prolactin response to suckling than normal-weight women at 48 hours but not at day 7. In multivariate analyses, overweight/obesity, primiparity, and birth weight were negatively associated with the prolactin response to suckling at 48 hours. After adjustment for confounding by time since delivery and the duration of the nursing episode, only overweight/obesity remained a significant negative predictor of prolactin response to suckling at day 7. Concentrations of progesterone decreased dramatically from 48 hours to 7 days postpartum but did not differ between normal weight and overweight/obese women at either time. In addition, the decreases in progesterone concentrations from 48 hours to 7 days postpartum did not differ between the prepregnant BMI groups. CONCLUSION: The unique and important finding from this study is that overweight/obese women had a lower prolactin response to suckling. This would be expected to compromise the ability of overweight/obese women to produce milk and, over time, could lead to premature cessation of lactation. These findings are important because, during our observation period (just before and after lactogenesis II, the time of onset of copious milk secretion), the prolactin response to suckling is more important for milk production than it is later in lactation. We have previously shown that a high proportion of the overweight and obese women in women in this population who give up on breastfeeding do so at this time. This finding thus provides evidence of a biological basis for this association, and additional study of it is likely to be informative. We postulated that there would be consistently higher progesterone concentrations in the early postpartum period among obese compared with normal weight women because adipose tissue is an extraplacental source of this hormone. This hypothesis was not supported in this study because there were no significant differences between normal-weight and overweight/obese women in progesterone concentrations at either 48 hours or 7 days postpartum. The values that we observed at these times were similar to those reported by others in the early postpartum period. The findings from this study add plausibility to our observation that initiation, not just duration of breastfeeding, is negatively affected by maternal overweight/obesity. Although women should begin pregnancy at a healthy weight and gain reasonably during gestation, not all will. Pediatricians can help overweight/obese women to succeed at breastfeeding by targeting them for contact with a lactation consultant before discharge from the hospital to be sure that they have received optimal advice on breastfeeding techniques. In addition, early contact with the mother after discharge--by calling her at home to offer her support and counseling for breastfeeding, by scheduling the first pediatric visit earlier than for other patients, or by enlisting the assistance of public health nurses for a home visit if this is possible--would help overweight/obese women to continue to breastfeed. Being overweight or obese is negatively associated with the prolactin response to suckling in the first week postpartum and, thus, may contribute to early lactation failure. PMID- 15121991 TI - The genetics of autism. AB - Autism is a complex, behaviorally defined, static disorder of the immature brain that is of great concern to the practicing pediatrician because of an astonishing 556% reported increase in pediatric prevalence between 1991 and 1997, to a prevalence higher than that of spina bifida, cancer, or Down syndrome. This jump is probably attributable to heightened awareness and changing diagnostic criteria rather than to new environmental influences. Autism is not a disease but a syndrome with multiple nongenetic and genetic causes. By autism (the autistic spectrum disorders [ASDs]), we mean the wide spectrum of developmental disorders characterized by impairments in 3 behavioral domains: 1) social interaction; 2) language, communication, and imaginative play; and 3) range of interests and activities. Autism corresponds in this article to pervasive developmental disorder (PDD) of the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition and International Classification of Diseases, Tenth Revision. Except for Rett syndrome--attributable in most affected individuals to mutations of the methyl-CpG-binding protein 2 (MeCP2) gene--the other PDD subtypes (autistic disorder, Asperger disorder, disintegrative disorder, and PDD Not Otherwise Specified [PDD-NOS]) are not linked to any particular genetic or nongenetic cause. Review of 2 major textbooks on autism and of papers published between 1961 and 2003 yields convincing evidence for multiple interacting genetic factors as the main causative determinants of autism. Epidemiologic studies indicate that environmental factors such as toxic exposures, teratogens, perinatal insults, and prenatal infections such as rubella and cytomegalovirus account for few cases. These studies fail to confirm that immunizations with the measles-mumps-rubella vaccine are responsible for the surge in autism. Epilepsy, the medical condition most highly associated with autism, has equally complex genetic/nongenetic (but mostly unknown) causes. Autism is frequent in tuberous sclerosis complex and fragile X syndrome, but these 2 disorders account for but a small minority of cases. Currently, diagnosable medical conditions, cytogenetic abnormalities, and single-gene defects (eg, tuberous sclerosis complex, fragile X syndrome, and other rare diseases) together account for <10% of cases. There is convincing evidence that "idiopathic" autism is a heritable disorder. Epidemiologic studies report an ASD prevalence of approximately 3 to 6/1000, with a male to female ratio of 3:1. This skewed ratio remains unexplained: despite the contribution of a few well characterized X-linked disorders, male-to-male transmission in a number of families rules out X-linkage as the prevailing mode of inheritance. The recurrence rate in siblings of affected children is approximately 2% to 8%, much higher than the prevalence rate in the general population but much lower than in single-gene diseases. Twin studies reported 60% concordance for classic autism in monozygotic (MZ) twins versus 0 in dizygotic (DZ) twins, the higher MZ concordance attesting to genetic inheritance as the predominant causative agent. Reevaluation for a broader autistic phenotype that included communication and social disorders increased concordance remarkably from 60% to 92% in MZ twins and from 0% to 10% in DZ pairs. This suggests that interactions between multiple genes cause "idiopathic" autism but that epigenetic factors and exposure to environmental modifiers may contribute to variable expression of autism-related traits. The identity and number of genes involved remain unknown. The wide phenotypic variability of the ASDs likely reflects the interaction of multiple genes within an individual's genome and the existence of distinct genes and gene combinations among those affected. There are 3 main approaches to identifying genetic loci, chromosomal regions likely to contain relevant genes: 1) whole genome screens, searching for linkage of autism to shared genetic markers in populations of multiplex families (families with >1 affected family member; 2) cytogenetic studies that may guide molecular studies by pointing to relevant inherited or de novo chromosomal abnormalities in affected individuals and their families; and 3) evaluation of candidate genes known to affect brain development in these significantly linked regions or, alternatively, linkage of candidate genes selected a priori because of their presumptive contribution to the pathogenesis of autism. Data from whole-genome screens in multiplex families suggest interactions of at least 10 genes in the causation of autism. Thus far, a putative speech and language region at 7q31-q33 seems most strongly linked to autism, with linkages to multiple other loci under investigation. Cytogenetic abnormalities at the 15q11-q13 locus are fairly frequent in people with autism, and a "chromosome 15 phenotype" was described in individuals with chromosome 15 duplications. Among other candidate genes are the FOXP2, RAY1/ST7, IMMP2L, and RELN genes at 7q22-q33 and the GABA(A) receptor subunit and UBE3A genes on chromosome 15q11-q13. Variant alleles of the serotonin transporter gene (5-HTT) on 17q11-q12 are more frequent in individuals with autism than in nonautistic populations. In addition, animal models and linkage data from genome screens implicate the oxytocin receptor at 3p25-p26. Most pediatricians will have 1 or more children with this disorder in their practices. They must diagnose ASD expeditiously because early intervention increases its effectiveness. Children with dysmorphic features, congenital anomalies, mental retardation, or family members with developmental disorders are those most likely to benefit from extensive medical testing and genetic consultation. The yield of testing is much less in high-functioning children with a normal appearance and IQ and moderate social and language impairments. Genetic counseling justifies testing, but until autism genes are identified and their functions are understood, prenatal diagnosis will exist only for the rare cases ascribable to single-gene defects or overt chromosomal abnormalities. Parents who wish to have more children must be told of their increased statistical risk. It is crucial for pediatricians to try to involve families with multiple affected members in formal research projects, as family studies are key to unraveling the causes and pathogenesis of autism. Parents need to understand that they and their affected children are the only available sources for identifying and studying the elusive genes responsible for autism. Future clinically useful insights and potential medications depend on identifying these genes and elucidating the influences of their products on brain development and physiology. PMID- 15121992 TI - Educating pediatricians on children's oral health: past, present, and future. AB - OBJECTIVE: The American Academy of Pediatrics (AAP) policy Oral Health Risk Assessment Timing and Establishment of the Dental Home encourages pediatricians to play an important role in the oral health of children. The purpose of this study was to determine how well pediatricians are prepared to play the AAP suggested role in children's oral health by examining the oral health content of their educational process. METHODS: This article reviews current medical education guidelines, programs, surveys, and pediatrician experiences in oral health training at the undergraduate, graduate, and continuing medical education levels. RESULTS: Although some medical schools, residency programs, and continuing medical education efforts do include oral health in their curricula, the practice is not widespread. Professional and oversight organizations such as the Association of American Medical Colleges, the Council on Medical Student Education in Pediatrics, Association of Pediatric Program Directors, AAP, and the pediatric Residency Review Committee do not include oral health in key guidelines and surveys of the medical education continuum. When surveyed, pediatricians state that the time spent on oral health education at each level of training is inadequate. CONCLUSIONS: Overall, the level of oral health training for pediatricians at the undergraduate, graduate, and continuing medical education levels is inadequate to provide pediatricians with the competencies required for the provision of quality oral health care to children. Despite this inadequacy, educational efforts are under way and various tools and guidelines are available in a few locations. In addition, numerous opportunities exist for the organizations responsible for pediatrician education to ensure that curricula are designed and adopted to achieve these competencies, faculty are trained to teach these competencies, and program accreditation and certification are closely linked to acquisition of these competencies. Ensuring the success of such efforts will require cooperation between many organizations and disciplines and a genuine commitment to improve pediatric education and medical care for children. PMID- 15121993 TI - Trichotillomania in childhood: case series and review. AB - Trichotillomania is a relatively common cause of childhood alopecia. We report our observations of 10 children with trichotillomania seen over a 2-year period at Texas Children's Hospital. Patient ages ranged from 9 to 14 years (mean: 11.3 years) with an equal gender ratio. The duration of hair-pulling ranged from 1 month to 10 years (median: 4.6 months). The scalp alone was affected in 8 cases, the scalp and eyelashes in 1 case, and the eyelashes alone in 1 case. The frontal scalp and vertex were the most common sites affected. Associated findings included nail-biting in 2 cases, "picking" of the skin in 1 case, and headaches in another case. Noted precipitating factors in 3 patients included "stress" at home and school. Associated psychopathology included major depression in 1 case, attention-deficit/hyperactivity disorder in 1 case, and an "anxious and nervous personality" in 1 case. The most important differential diagnosis to exclude from trichotillomania is alopecia areata, which was seen concomitantly in 1 patient and preceded the onset of hair-pulling by 11 months. Eight patients were referred to a child psychologist for additional management, of which 2 were subsequently treated with antidepressant medication. Trichotillomania is a disorder of multifaceted pathology, and an interdisciplinary approach to management is often helpful. The common prepubertal age of onset provides an important opportunity for the pediatrician to lend support to affected patients and their families. PMID- 15121994 TI - Grand mal seizure in a child 30 minutes after Cyclogyl (cyclopentolate hydrochloride) and 10% Neo-Synephrine (phenylephrine hydrochloride) eye drops were instilled. AB - A grand mal seizure is an unexpected, rare adverse event in a child receiving eye drops to dilate the pupils for an eye examination. A case is reported of a convulsion in a 23-month-old boy after he received Cyclogyl (cyclopentolate) and Neo-Synephrine (phenylephrine) eye drops before ophthalmoscopy. His serum sodium was 125 mEq/L, and he had low plasma pseudocholinesterase activity. Children exposed to organophosphate insecticides and other pseudocholinesterase inhibitors may be at risk for cyclopentolate toxicity. PMID- 15121995 TI - Unilateral negative-pressure pulmonary edema in an infant during bronchoscopy. AB - We report here a case of a 3-week-old infant with a lung disease of unknown origin who underwent diagnostic fiber-optic bronchoscopy. During the procedure, she developed unilateral pulmonary edema. Factors that contributed to this complication are discussed along with means to reduce its occurrence. PMID- 15121996 TI - Intestinal pseudoobstruction in Kawasaki disease. AB - Intestinal pseudoobstruction is an uncommon but important manifestation of Kawasaki disease. Its occurrence at the onset or during the course of the disease may confuse the clinical picture and cause delay in diagnosis and treatment. This delay may be responsible for the high rate of coronary artery abnormalities that have been reported in patients with this complication. We suggest that Kawasaki disease be considered in the differential diagnosis of any child presenting with intestinal pseudoobstruction and fever without definable cause. PMID- 15121997 TI - Coronary event secondary prevention with statins irrespective of LDL-cholesterol. AB - OBJECTIVE: To review the evidence for statin secondary prevention of coronary artery disease in patients with near-optimal or optimal low-density lipoprotein cholesterol (LDL-C). DATA SOURCES: A MEDLINE search (1966-October 2003) was conducted using the search terms HMG-CoA reductase inhibitor, statins, coronary disease, post-myocardial infarction, and average cholesterol. DATA SYNTHESIS: Secondary prevention trials enrolling subjects with near-optimal (<130 mg/dL) or optimal (<100 mg/dL) baseline LDL-C were included. Early statin secondary prevention studies suggested attenuated benefit, but more recent trials challenge this finding. CONCLUSIONS: Statin secondary prevention of coronary artery disease in patients near goal LDL-C is controversial, but recent trial results show promise. PMID- 15121998 TI - Diabetes mellitus non-glucose monitoring: point-of-care testing. AB - OBJECTIVE: To review and evaluate reimbursable point-of-care testing devices yielding immediate results, other than glucometers, that are available to evaluate and monitor diabetes and its complications and to describe how pharmacists may use these devices. DATA SOURCES: A MEDLINE search (1966-March 2003) was performed using the following search terms: point-of-care systems, clinical diabetes monitoring, decision support systems, glycosylated hemoglobin, and microalbumin. Pertinent company and product Web sites and customer service departments were accessed for information about point-of-care devices and supplies. STUDY SELECTION AND DATA EXTRACTION: All descriptive, evaluative, and comparative articles and product information were reviewed, and relevant information was included. DATA SYNTHESIS: Diabetes mellitus is a complex, chronic metabolic disease that is a challenging management problem and requires routine monitoring for disease control and screening for complications. Point-of-care tests are available to monitor hemoglobin A(1c), glucose, fructosamine, ketones, lipid profiles, urinary microalbumin concentrations, and alanine aminotransferase concentrations. Many of these tests are Clinical Laboratory Improvement Amendments (CLIA)-waived and, therefore, practical for pharmacists to use in a variety of settings. Tests for measuring sensation are also discussed. Pharmacists should consider each of these tests in the establishment of a comprehensive diabetes care service. CONCLUSIONS: The availability of many new point-of-care testing methods creates new opportunities for pharmacists to monitor drug therapy and screen for complications in patients with diabetes. Reimbursement is possible since many of these tests are CLIA-waived. PMID- 15121999 TI - Emtricitabine: a once-daily nucleoside reverse transcriptase inhibitor. AB - OBJECTIVE: To review the pharmacology, virology, pharmacokinetics, safety, and efficacy of the nucleoside reverse transcriptase inhibitor (NRTI) emtricitabine. DATA SOURCES: English-language reports were accessed using MEDLINE (1966-June 2003) and the Iowa Drug Information Service database (1966-June 2003) using emtricitabine and Coviracil as key words. (Coviracil was the proposed trade name for the product prior to approval.) The Internet was also searched using the terms HIV/AIDS conferences, then emtricitabine within the conference proceedings. STUDY SELECTION AND DATA EXTRACTION: Abstracts, posters, and oral presentations from scientific conferences, both published and unpublished, were included. Preference was given to published controlled trials. Studies providing a description of the pharmacology, virology, effectiveness, safety, or pharmacokinetics of emtricitabine were used in this review. DATA SYNTHESIS: Emtricitabine is an NRTI used to treat HIV-1 infection. Once-daily administration can decrease pill burden and potentially increase adherence to multidrug HIV therapy. Further, emtricitabine has shown equivalent or improved outcomes compared with lamivudine and stavudine. CONCLUSIONS: Emtricitabine is a safe and effective option for HIV-1 infection in adults as part of a multidrug regimen. It may be a better alternative than lamivudine for once-daily therapy because of its extended intracellular half-life and better than lamivudine and stavudine because of a possibly decreased potential for drug resistance. PMID- 15122000 TI - Continuous subcutaneous infusion of opiates at end-of-life. AB - OBJECTIVE: To review pertinent controlled trials using the continuous subcutaneous infusion of opioids (CSIO) at end-of-life and offer insight to pharmacists and clinicians into the appropriate use of this route of administration. DATA SOURCES: A MEDLINE search for information regarding the subcutaneous administration of opioids in terminally ill patients (1975-December 2002) was conducted using the key words subcutaneous, narcotics, morphine, hydromorphone, fentanyl, pain, hospices, and palliative care. Additional references were located through review of bibliographies of the articles cited. Case reports and postsurgical studies were excluded. Searches were limited to English-language studies using humans. STUDY SELECTION AND DATA EXTRACTION: Experimental and observational studies were evaluated, using prospective trials as the evidence base for conclusions and including pertinent retrospective trials as they relate to the subcutaneous infusion of opioids at end-of-life. DATA SYNTHESIS: CSIO is effective and safe for use in terminal illness. Appropriate situations for consideration of CSIO are when difficulties arise in using the oral route, standard oral opiate therapy has failed adequate trials, the patient has limited intravenous access, adequate supervision of the CSIO is present, and CSIO will not unduly limit the functional activity of the patient. CONCLUSIONS: CSIO has a proven role in the management of pain at end-of-life. CSIO should not be considered the first route for administration of opiates, but does offer distinct advantages in the appropriate setting. CSIO continues to be a choice for end-of-life patients and is gradually becoming a standard practice in palliative medicine. PMID- 15122001 TI - Ursodeoxycholic acid for terbinafine-induced toxic hepatitis. PMID- 15122002 TI - Muehrcke's lines on nails after cyclophosphamide/adriamycin/fluorouracil. PMID- 15122003 TI - Evaluation of a physician-focused educational intervention on medicaid children with asthma. AB - BACKGROUND: The 1990 Omnibus Budget Reconciliation Act mandated drug utilization review in response to inappropriate drug use. In the Pennsylvania Medicaid program, pediatric asthma is associated with high healthcare utilization and cost. OBJECTIVE: To determine the effects of a physician-focused educational intervention on asthma drug use and healthcare utilization. METHODS: Pre- and postintervention comparison design was used in children 5-18 years of age who were enrolled in the Pennsylvania Medicaid fee-for-service program from July 1, 1998, to March 31, 1999 (preintervention), and July 1, 1999, to March 31, 2000 (postintervention). The intervention packet included patients' drug profiles, medical history, monograph with national asthma management guidelines, and patient education materials to physicians. Main outcome measures are changes in asthma drug utilization among high-users of short-acting beta(2)-agonists (SAB). RESULTS: The intervention focused on 2 asthma drug use criteria: (1) high-use of quick-relief medication and (2) use of salmeterol without the availability of a quick-relief medication. The intervention reduced quick-relief medication use by 26% among patients with higher use without significant changes in long-term control drugs. In addition, 82% of the recipients evaluated had a positive change in salmeterol utilization as either having an SAB inhaler added after the intervention or salmeterol discontinued after the intervention. There was no significant change in asthma-related emergency department visits or hospitalizations. CONCLUSIONS: Although the physician responders agreed on the usefulness of the educational materials, the results suggest that the intervention had limited success in improving the pharmacologic management and no effect on the health outcomes. We believe that mailed educational materials to physicians can be effective to change prescribing behavior; however, a more multifaceted intervention may be necessary to improve health outcomes. PMID- 15122004 TI - Incidence of hepatotoxicity due to antitubercular medicines and assessment of risk factors. AB - BACKGROUND: Antitubercular drugs cause derangement of hepatic function revealed by clinical examination and abnormal liver function test results. Potential hepatotoxicity of some of the first-line antitubercular agents remains a problem, especially during the initial period of treatment. OBJECTIVE: To determine the incidence of antitubercular drug-induced hepatotoxicity in a Nepalese urban population and assess the risk factors. METHOD: Fifty patients diagnosed with active tuberculosis infection with normal pretreatment liver function were monitored clinically as well as biochemically in a prospective cohort analysis. RESULTS: Antitubercular drugs were found to be associated with derangement of hepatic function, resulting in elevation of liver enzymes to a variable extent (t = -4.550, p < 0.01 for aspartate aminotransferase [AST]; t = -5.467, p < 0.01 for alanine aminotransferase [ALT] at 95% CI). Thirty-eight percent of patients had 2 times and 30% had >3 times elevation of ALT. Similarly, 40% and 29% of patients showed 2 and >3 times elevation of the AST level, respectively. Four patients (8%) developed drug-induced hepatotoxicity. Jaundice was the presenting symptom in all patients. The time interval for onset of hepatotoxicity after initiation of therapy was 12-60 days (median 28). Antitubercular drug-induced hepatotoxicity was found more often in younger patients (6% vs 2%; p = 0.368, OR 2.75). Female gender was also a higher risk (p = 0.219, OR 4.2). Most patients who had developed hepatitis were diagnosed per sputum-smear positive reactions. Nutritional status, assessed by body mass index and serum albumin level, was the next predisposing factor. CONCLUSIONS: A finding of an 8% incidence of hepatotoxicity is considerably high. Risk factors of hepatotoxicity included female gender, disease extent, and poor nutritional status. Timely detection and temporary withdrawal of the offending agent can completely cure antitubercular drug-induced hepatotoxicity. PMID- 15122005 TI - Intrathecal amikacin for the treatment of pseudomonal meningitis. AB - OBJECTIVE: To report a case of gram-negative bacillary meningitis (GNBM) secondary to multidrug-resistant Pseudomonas aeruginosa that was treated with intravenous meropenem and intrathecal and intravenous amikacin. CASE SUMMARY: A 76-year-old Arabic woman with previous placement of an extraventricular device developed meningitis secondary to P. aeruginosa as a result of a previous pneumonia. The patient was treated with intravenous meropenem and amikacin, with the addition of intrathecal amikacin, until cerebrospinal cultures remained negative for 18 days. She did not experience any adverse effects as a result of the administration of the intrathecal amikacin. Although the meningitis subsequently resolved, the patient eventually died due to Candida glabrata fungemia. DISCUSSION: Dual therapy is recommended for patients with P. aeruginosa meningitis. In our patient, the increasing resistance to imipenem and resistance to all other potential antibiotics resulted in the use of an alternative administration technique that has not been well documented in recent literature. CONCLUSIONS: In patients who have GNBM due to P. aeruginosa, the combination of intrathecal and intravenous amikacin may be an option for therapy, especially when clinical options are limited by resistance, severity of illness, and location of the infection. More information is required and further study is needed on this topic. PMID- 15122006 TI - Pulmonary edema and possible pneumonitis associated with celecoxib. PMID- 15122007 TI - Progress toward measles elimination--region of the Americas, 2002-2003. PMID- 15122008 TI - Usefulness of oral cyanocobalamin therapy in severe hematologic manifestations related to vitamin B12 deficiency. PMID- 15122009 TI - Identification of a new chloroplast carbonic anhydrase in Chlamydomonas reinhardtii. AB - Carbonic anhydrases (CA) are zinc-containing metalloenzymes that catalyze the reversible hydration of CO2. The three evolutionarily unrelated families of CAs are designated alpha-, beta-, and gamma-CA. Aquatic photosynthetic organisms have evolved different forms of CO2 concentrating mechanisms (CCMs) to aid Rubisco in capturing CO2 from the surrounding environment. One aspect of all CCMs is the critical roles played by various specially localized extracellular and intracellular CAs. Five CAs have previously been identified in Chlamydomonas reinhardtii, a green alga with a well-studied CCM. Here we identify a sixth gene encoding a beta-type CA. This new beta-CA, designated Cah6, is distinct from the two mitochondrial beta-CAs in C. reinhardtii. Nucleotide sequence data show that the Cah6 cDNA contains an open reading frame encoding a polypeptide of 264 amino acids with a leader sequence likely targeting the protein to the chloroplast stroma. We have fused the Cah6 open reading frame to the coding sequence of maltose-binding protein in a pMal expression vector. The purified recombinant fusion protein is active and was used to partially characterize the Cah6 protein. The purified recombinant fusion protein was cleaved with protease Factor Xa to separate Cah6 from the maltose-binding protein and the purified Cah6 protein was used to raise an antibody. Western blots, immunolocalization studies, and northern blots collectively indicated that Cah6 is constitutively expressed in the stroma of chloroplasts. A possible role for Cah6 in the CCM of C. reinhardtii is proposed. PMID- 15122010 TI - Contribution of different carbon sources to isoprene biosynthesis in poplar leaves. AB - This study was performed to test if alternative carbon sources besides recently photosynthetically fixed CO2 are used for isoprene formation in the leaves of young poplar (Populus x canescens) trees. In a 13CO2 atmosphere under steady state conditions, only about 75% of isoprene became 13C labeled within minutes. A considerable part of the unlabeled carbon may be derived from xylem transported carbohydrates, as may be shown by feeding leaves with [U-13C]Glc. As a consequence of this treatment approximately 8% to 10% of the carbon emitted as isoprene was 13C labeled. In order to identify further carbon sources, poplar leaves were depleted of leaf internal carbon pools and the carbon pools were refilled with 13C labeled carbon by exposure to 13CO2. Results from this treatment showed that about 30% of isoprene carbon became 13C labeled, clearly suggesting that, in addition to xylem transported carbon and CO2, leaf internal carbon pools, e.g. starch, are used for isoprene formation. This use was even increased when net assimilation was reduced, for example by abscisic acid application. The data provide clear evidence of a dynamic exchange of carbon between different cellular precursors for isoprene biosynthesis, and an increasing importance of these alternative carbon pools under conditions of limited photosynthesis. Feeding [1,2-13C]Glc and [3-13C]Glc to leaves via the xylem suggested that alternative carbon sources are probably derived from cytosolic pyruvate/phosphoenolpyruvate equivalents and incorporated into isoprene according to the predicted cleavage of the 3-C position of pyruvate during the initial step of the plastidic deoxyxylulose-5-phosphate pathway. PMID- 15122011 TI - Copper transport across pea thylakoid membranes. AB - The initial rate of Cu2+ movement across the thylakoid membrane of pea (Pisum sativum) chloroplasts was directly measured by stopped-flow spectrofluorometry using membranes loaded with the Cu(2+)-sensitive fluorophore Phen Green SK. Cu2+ transport was rapid, reaching completion within 0.5 s. The initial rate of uptake was dependent upon Cu2+ concentration and saturated at about 0.6 microm total Cu2+. Cu2+ uptake was maximal at a thylakoid lumen pH of 7.0. Cu2+ transport was inhibited by Zn2+ but was largely unaffected by Mn2+ and Cu+. Zn2+ inhibited Cu2+ transport to a maximum of 60%, indicating that there may be more than one transporter for copper in pea thylakoid membranes. PMID- 15122012 TI - Structure and expression profile of the Arabidopsis PHO1 gene family indicates a broad role in inorganic phosphate homeostasis. AB - PHO1 has been recently identified as a protein involved in the loading of inorganic phosphate into the xylem of roots in Arabidopsis. The genome of Arabidopsis contains 11 members of the PHO1 gene family. The cDNAs of all PHO1 homologs have been cloned and sequenced. All proteins have the same topology and harbor a SPX tripartite domain in the N-terminal hydrophilic portion and an EXS domain in the C-terminal hydrophobic portion. The SPX and EXS domains have been identified in yeast (Saccharomyces cerevisiae) proteins involved in either phosphate transport or sensing or in sorting proteins to endomembranes. The Arabidopsis genome contains additional proteins of unknown function containing either a SPX or an EXS domain. Phylogenetic analysis indicated that the PHO1 family is subdivided into at least three clusters. Reverse transcription-PCR revealed a broad pattern of expression in leaves, roots, stems, and flowers for most genes, although two genes are expressed exclusively in flowers. Analysis of the activity of the promoter of all PHO1 homologs using promoter-beta glucuronidase fusions revealed a predominant expression in the vascular tissues of roots, leaves, stems, or flowers. beta-Glucuronidase expression is also detected for several promoters in nonvascular tissue, including hydathodes, trichomes, root tip, root cortical/epidermal cells, and pollen grains. The expression pattern of PHO1 homologs indicates a likely role of the PHO1 proteins not only in the transfer of phosphate to the vascular cylinder of various tissues but also in the acquisition of phosphate into cells, such as pollen or root epidermal/cortical cells. PMID- 15122013 TI - Biosynthesis of the nitrile glucosides rhodiocyanoside A and D and the cyanogenic glucosides lotaustralin and linamarin in Lotus japonicus. AB - Lotus japonicus was shown to contain the two nitrile glucosides rhodiocyanoside A and rhodiocyanoside D as well as the cyanogenic glucosides linamarin and lotaustralin. The content of cyanogenic and nitrile glucosides in L. japonicus depends on plant developmental stage and tissue. The cyanide potential is highest in young seedlings and in apical leaves of mature plants. Roots and seeds are acyanogenic. Biosynthetic studies using radioisotopes demonstrated that lotaustralin, rhodiocyanoside A, and rhodiocyanoside D are derived from the amino acid l-Ile, whereas linamarin is derived from Val. In silico homology searches identified two cytochromes P450 designated CYP79D3 and CYP79D4 in L. japonicus. The two cytochromes P450 are 94% identical at the amino acid level and both catalyze the conversion of Val and Ile to the corresponding aldoximes in biosynthesis of cyanogenic glucosides and nitrile glucosides in L. japonicus. CYP79D3 and CYP79D4 are differentially expressed. CYP79D3 is exclusively expressed in aerial parts and CYP79D4 in roots. Recombinantly expressed CYP79D3 and CYP79D4 in yeast cells showed higher catalytic efficiency with l-Ile as substrate than with l-Val, in agreement with lotaustralin and rhodiocyanoside A and D being the major cyanogenic and nitrile glucosides in L. japonicus. Ectopic expression of CYP79D2 from cassava (Manihot esculenta Crantz.) in L. japonicus resulted in a 5- to 20-fold increase of linamarin content, whereas the relative amounts of lotaustralin and rhodiocyanoside A/D were unaltered. PMID- 15122014 TI - Rapid genome evolution revealed by comparative sequence analysis of orthologous regions from four triticeae genomes. AB - Bread wheat (Triticum aestivum) is an allohexaploid species, consisting of three subgenomes (A, B, and D). To study the molecular evolution of these closely related genomes, we compared the sequence of a 307-kb physical contig covering the high molecular weight (HMW)-glutenin locus from the A genome of durum wheat (Triticum turgidum, AABB) with the orthologous regions from the B genome of the same wheat and the D genome of the diploid wheat Aegilops tauschii (Anderson et al., 2003; Kong et al., 2004). Although gene colinearity appears to be retained, four out of six genes including the two paralogous HMW-glutenin genes are disrupted in the orthologous region of the A genome. Mechanisms involved in gene disruption in the A genome include retroelement insertions, sequence deletions, and mutations causing in-frame stop codons in the coding sequences. Comparative sequence analysis also revealed that sequences in the colinear intergenic regions of these different genomes were generally not conserved. The rapid genome evolution in these regions is attributable mainly to the large number of retrotransposon insertions that occurred after the divergence of the three wheat genomes. Our comparative studies indicate that the B genome diverged prior to the separation of the A and D genomes. Furthermore, sequence comparison of two distinct types of allelic variations at the HMW-glutenin loci in the A genomes of different hexaploid wheat cultivars with the A genome locus of durum wheat indicates that hexaploid wheat may have more than one tetraploid ancestor. PMID- 15122015 TI - Calcium interacts with antifreeze proteins and chitinase from cold-acclimated winter rye. AB - During cold acclimation, winter rye (Secale cereale) plants accumulate pathogenesis-related proteins that are also antifreeze proteins (AFPs) because they adsorb onto ice and inhibit its growth. Although they promote winter survival in planta, these dual-function AFPs proteins lose activity when stored at subzero temperatures in vitro, so we examined their stability in solutions containing CaCl2, MgCl2, or NaCl. Antifreeze activity was unaffected by salts before freezing, but decreased after freezing and thawing in CaCl2 and was recovered by adding a chelator. Ca2+ enhanced chitinase activity 3- to 5-fold in unfrozen samples, although hydrolytic activity also decreased after freezing and thawing in CaCl2. Native PAGE, circular dichroism, and Trp fluorescence experiments showed that the AFPs partially unfold after freezing and thawing, but they fold more compactly or aggregate in CaCl2. Ruthenium red, which binds to Ca(2+)-binding sites, readily stained AFPs in the absence of Ca2+, but less stain was visible after freezing and thawing AFPs in CaCl2. We conclude that the structure of AFPs changes during freezing and thawing, creating new Ca(2+) binding sites. Once Ca2+ binds to those sites, antifreeze activity, chitinase activity and ruthenium red binding are all inhibited. Because free Ca2+ concentrations are typically low in the apoplast, antifreeze activity is probably stable to freezing and thawing in planta. Ca2+ may regulate chitinase activity if concentrations are increased locally by release from pectin or interaction with Ca(2+)-binding proteins. Furthermore, antifreeze activity can be easily maintained in vitro by including a chelator during frozen storage. PMID- 15122016 TI - Differential timing of spider mite-induced direct and indirect defenses in tomato plants. AB - Through a combined metabolomics and transcriptomics approach we analyzed the events that took place during the first 5 d of infesting intact tomato (Lycopersicon esculentum) plants with spider mites (Tetranychus urticae). Although the spider mites had caused little visible damage to the leaves after 1 d, they had already induced direct defense responses. For example, proteinase inhibitor activity had doubled and the transcription of genes involved in jasmonate-, salicylate-, and ethylene-regulated defenses had been activated. On day four, proteinase inhibitor activity and particularly transcript levels of salicylate-regulated genes were still maintained. In addition, genes involved in phospholipid metabolism were up-regulated on day one and those in the secondary metabolism on day four. Although transcriptional up-regulation of the enzymes involved in the biosynthesis of monoterpenes and diterpenes already occurred on day one, a significant increase in the emission of volatile terpenoids was delayed until day four. This increase in volatile production coincided with the increased olfactory preference of predatory mites (Phytoseiulus persimilis) for infested plants. Our results indicate that tomato activates its indirect defenses (volatile production) to complement the direct defense response against spider mites. PMID- 15122017 TI - Production of coumaroylserotonin and feruloylserotonin in transgenic rice expressing pepper hydroxycinnamoyl-coenzyme A:serotonin N (hydroxycinnamoyl)transferase. AB - Transgenic rice (Oryza sativa) plants were engineered to express a N (hydroxycinnamoyl)transferase from pepper (Capsicum annuum), which has been shown to have hydroxycinnamoyl-CoA:tyramine N-(hydroxycinnamoyl)transferase activity, a key enzyme in the synthesis of hydroxycinnamic acid amides, under the control of constitutive maize (Zea mays) ubiquitin promoter. The transgenic rice plants require foliar application of amines to support synthesis of hydroxycinnamic acid amides, suggestive of limiting amine substrates in rice shoots. In addition, when T2 homozygous transgenic rice plants were grown in the presence of amines or phenolic acids, two novel compounds were exclusively identified in the leaves of the transgenic plants. These compounds eluted earlier than p-coumaroyltyramine and feruloyltyramine during HPLC chromatography and were identified as p coumaroylserotonin and feruloylserotonin by liquid chromatography/mass spectrometry and other methods. To test whether the unpredicted production of serotonin derivatives is associated with the pepper N (hydroxycinnamoyl)transferase, the substrate specificity of the pepper enzyme was analyzed again. Purified recombinant pepper N-(hydroxycinnamoyl)transferase exhibited a serotonin N-hydroxycinnamoyltransferase (SHT) activity, synthesized p coumaroylserotonin and feruloylserotonin in vitro, and demonstrated a low K(m) for serotonin. SHT activity was inhibited by 10 to 50 mm tyramine. In addition, SHT activity was predominantly found in the root tissues of wild-type rice in parallel with the synthesis of serotonin derivatives, suggesting that serotonin derivatives are synthesized in the root of rice. This is the first report of SHT activity and the first demonstration, to our knowledge, that serotonin derivatives can be overproduced in vivo in transgenic rice plants that express serotonin N-(hydroxycinnamoyl)transferase. PMID- 15122018 TI - Nitric oxide mediates the indole acetic acid induction activation of a mitogen activated protein kinase cascade involved in adventitious root development. AB - Recently, it was demonstrated that nitric oxide (NO) and cGMP are involved in the auxin response during the adventitious rooting process in cucumber (Cucumis sativus; Pagnussat et al., 2002, 2003). However, not much is known about the complex molecular network operating during the cell proliferation and morphogenesis triggered by auxins and NO in that process. Anatomical studies showed that formation of adventitious root primordia was clearly detected in indole acetic acid (IAA)- and NO-treated cucumber explants, while neither cell proliferation nor differentiation into root primordia could be observed in control explants 3 d after primary root was removed. In order to go further with signal transduction mechanisms that operate during IAA- and NO-induced adventitious root formation, experiments were designed to test the involvement of a mitogen-activated protein kinase (MAPK) cascade in that process. Cucumber explants were treated with the NO-donor sodium nitroprusside (SNP) or with SNP plus the specific NO-scavenger cPTIO. Protein extracts from those explants were assayed for protein kinase (PK) activity by using myelin basic protein (MBP) as substrate in both in vitro and in-gel assays. The activation of a PK of approximately 48 kD could be detected 1 d after NO treatment with a maximal activation after 3 d of treatment. In control explants, a PK activity was detected only after 4 d of treatment. The MBP-kinase activity was also detected in extracts from IAA-treated explants, while no signal was observed in IAA + cPTIO treatments. The PK activity could be inhibited by the cell-permeable MAPK kinase inhibitor PD098059, suggesting that the NO-dependent MBP-kinase activity is a MAPK. Furthermore, when PD098059 was administered to explants treated with SNP or IAA, it produced a delay in root emergence and a dose-dependent reduction in root number. Altogether, our results suggest that a MAPK signaling cascade is activated during the adventitious rooting process induced by IAA in a NO-mediated but cGMP-independent pathway. The activation of MAPKs is discussed in relation to the cell responses modulating mitotic process. PMID- 15122019 TI - Cloning and overproduction of gibberellin 3-oxidase in hybrid aspen trees. Effects on gibberellin homeostasis and development. AB - To broaden our understanding of gibberellin (GA) biosynthesis and the mechanism whereby GA homeostasis is maintained in plants, we have investigated the degree to which the enzyme GA 3-oxidase (GA3ox) limits the formation of bioactive GAs in elongating shoots of hybrid aspen (Populus tremula x Populus tremuloides). We describe the cloning of a hybrid aspen GA3ox and its functional characterization, which confirmed that it has 3beta-hydroxylation activity and more efficiently converts GA9 to GA4 than GA20 to GA1. To complement previous studies, in which transgenic GA 20-oxidase (GA20ox) overexpressers were found to produce 20-fold higher bioactive GA levels and subsequently grew faster than wild-type plants, we overexpressed an Arabidopsis GA3ox in hybrid aspen. The generated GA3ox overexpresser lines had increased 3beta-hydroxylation activity but exhibited no major changes in morphology. The nearly unaltered growth pattern was associated with relatively small changes in GA1 and GA4 levels, although tissue-dependent differences were observed. The absence of increases in bioactive GA levels did not appear to be due to feedback or feed-forward regulation of dioxygenase transcripts, according to semiquantitative reverse transcription polymerase chain reaction analysis of PttGA20ox1, PttGA3ox1, and two putative PttGA2ox genes. We conclude that 20-oxidation is the limiting step, rather than 3beta-hydroxylation, in the formation of GA1 and GA4 in elongating shoots of hybrid aspen, and that ectopic GA3ox expression alone cannot increase the flux toward bioactive GAs. Finally, several lines of evidence now suggest that GA4 has a more pivotal role in the tree hybrid aspen than previously believed. PMID- 15122020 TI - Analysis of nitric oxide signaling functions in tobacco cells challenged by the elicitor cryptogein. AB - Nitric oxide (NO) has recently emerged as an important cellular mediator in plant defense responses. However, elucidation of the biochemical mechanisms by which NO participates in this signaling pathway is still in its infancy. We previously demonstrated that cryptogein, an elicitor of tobacco defense responses, triggers a NO burst within minutes in epidermal sections from tobacco leaves (Nicotiana tabacum cv Xanthi). Here, we investigate the signaling events that mediate NO production, and analyze NO signaling activities in the cryptogein transduction pathway. Using flow cytometry and spectrofluorometry, we observed that cryptogein induced NO production in tobacco cell suspensions is sensitive to nitric oxide synthase inhibitors and may be catalyzed by variant P, a recently identified pathogen-inducible plant nitric oxide synthase. NO synthesis is tightly regulated by a signaling cascade involving Ca2+ influx and phosphorylation events. Using tobacco cells constitutively expressing the Ca2+ reporter apoaequorin in the cytosol, we have shown that NO participates in the cryptogein-mediated elevation of cytosolic free Ca2+ through the mobilization of Ca2+ from intracellular stores. The NO donor diethylamine NONOate promoted an increase in cytosolic free Ca2+ concentration, which was sensitive to intracellular Ca2+ channel inhibitors. Moreover, NO appears to be involved in the pathway(s) leading to the accumulation of transcripts encoding the heat shock protein TLHS-1, the ethylene-forming enzyme cEFE-26, and cell death. In contrast, NO does not act upstream of the elicitor-induced activation of mitogen-activated protein kinase, the opening of anion channels, nor expression of GST, LOX-1, PAL, and PR-3 genes. Collectively, our data indicate that NO is intimately involved in the signal transduction processes leading to cryptogein-induced defense responses. PMID- 15122022 TI - Extensive phenotypic variation in early flowering mutants of Arabidopsis. AB - Flowering time, the major regulatory transition of plant sequential development, is modulated by multiple endogenous and environmental factors. By phenotypic profiling of 80 early flowering mutants of Arabidopsis, we examine how mutational reduction of floral repression is associated with changes in phenotypic plasticity and stability. Flowering time measurements in mutants reveal deviations from the linear relationship between the number of leaves and number of days to bolting described for natural accessions and late flowering mutants. The deviations correspond to relative early bolting and relative late bolting phenotypes. Only a minority of mutants presents no detectable phenotypic variation. Mutants are characterized by a broad release of morphological pleiotropy under short days, with leaf characters being most variable. They also exhibit changes in phenotypic plasticity across environments for florigenic related responses, including the reaction to light and dark, photoperiodic behavior, and Suc sensitivity. Morphological pleiotropy and plasticity modifications are differentially distributed among mutants, resulting in a large diversity of multiple phenotypic changes. The pleiotropic effects observed may indicate that floral repression defects are linked to global developmental perturbations. This first, to our knowledge, extensive characterization of phenotypic variation in early flowering mutants correlates with the reports that most factors recruited in floral repression at the molecular genetic level correspond to ubiquitous regulators. We discuss the importance of functional ubiquity for floral repression with respect to robustness and flexibility of network biological systems. PMID- 15122023 TI - A comparison of rice chloroplast genomes. AB - Using high quality sequence reads extracted from our whole genome shotgun repository, we assembled two chloroplast genome sequences from two rice (Oryza sativa) varieties, one from 93-11 (a typical indica variety) and the other from PA64S (an indica-like variety with maternal origin of japonica), which are both parental varieties of the super-hybrid rice, LYP9. Based on the patterns of high sequence coverage, we partitioned chloroplast sequence variations into two classes, intravarietal and intersubspecific polymorphisms. Intravarietal polymorphisms refer to variations within 93-11 or PA64S. Intersubspecific polymorphisms were identified by comparing the major genotypes of the two subspecies represented by 93-11 and PA64S, respectively. Some of the minor genotypes occurring as intravarietal polymorphisms in one variety existed as major genotypes in the other subspecific variety, thus giving rise to intersubspecific polymorphisms. In our study, we found that the intersubspecific variations of 93-11 (indica) and PA64S (japonica) chloroplast genomes consisted of 72 single nucleotide polymorphisms and 27 insertions or deletions. The intersubspecific polymorphism rates between 93-11 and PA64S were 0.05% for single nucleotide polymorphisms and 0.02% for insertions or deletions, nearly 8 and 10 times lower than their respective nuclear genomes. Based on the total number of nucleotide substitutions between the two chloroplast genomes, we dated the divergence of indica and japonica chloroplast genomes as occurring approximately 86,000 to 200,000 years ago. PMID- 15122024 TI - Patterns of auxin distribution during gravitational induction of reaction wood in poplar and pine. AB - Gravistimulation of tree stems affects wood development by unilaterally inducing wood with modified properties, called reaction wood. Commonly, it also stimulates cambial growth on the reaction wood side. Numerous experiments involving applications of indole-3-acetic acid (IAA) or IAA-transport inhibitors have suggested that reaction wood is induced by a redistribution of IAA around the stem. However, in planta proof for this model is lacking. Therefore, we have mapped endogenous IAA distribution across the cambial region tissues in both aspen (Populus tremula, denoted poplar) and Scots pine (Pinus sylvestris) trees forming reaction wood, using tangential cryosectioning combined with sensitive gas chromatography-mass spectrometry analysis. Moreover, we have documented the kinetics of IAA during reaction wood induction in these species. Our analysis of endogenous IAA demonstrates that reaction wood is formed without any obvious alterations in IAA balance. This is in contrast to gravitropic responses in roots and shoots where a redistribution of IAA has been documented. It is also of interest that cambial growth on the tension wood side was stimulated without an increase in IAA. Taken together, our results suggest a role for signals other than IAA in the reaction wood response, or that the gravitational stimulus interacts with the IAA signal transduction pathway. PMID- 15122025 TI - Lysine metabolism is concurrently regulated by synthesis and catabolism in both reproductive and vegetative tissues. AB - The functional role of Lys catabolism in balancing Lys levels in plants has only been directly demonstrated in developing seeds. Seed-specific expression of a bacterial feedback-insensitive dihydrodipicolinate synthase (DHPS) in an Arabidopsis knockout mutant of the AtLKR/SDH gene that regulates Lys catabolism synergistically boosted Lys accumulation in mature seeds, but it also severely reduced the growth of seedlings derived from them. Here we further tested whether the inhibition of seedling growth was due to a negative physiological effect of excess Lys on seed maturation or to defective postgermination catabolism of Lys, which accumulated in the mature seeds. To address these questions, we coexpressed a bacterial DHPS gene with an RNAi construct of AtLKR/SDH, both under control of the same seed-specific promoter, to restrict Lys synthesis and catabolism to the developing seeds. Coexpression of these genes boosted seed Lys content and caused a significant, metabolically unanticipated increase in Met content, similarly to our previous report using plants expressing the bacterial DHPS on an AtLKR/SDH knockout background. However, postgermination seedling growth was significantly improved when the reduction of Lys catabolism was restricted to seed development, suggesting that defective postgermination Lys catabolism was responsible for inhibition of seedling growth in the AtLKR/SDH knockout plants expressing the bacterial DHPS gene in a seed-specific manner. Constitutive expression of the bacterial DHPS in the AtLKR/SDH knockout mutant boosted Lys levels in vegetative tissues in a similar manner to that observed in seeds, further demonstrating that Lys catabolism plays an important regulatory role in balancing Lys levels. PMID- 15122026 TI - Optospectroscopic detection of primary reactions associated with the graviperception of Phycomyces. Effects of micro- and hypergravity. AB - The graviperception of sporangiophores of the fungus Phycomyces blakesleeanus involves gravity-induced absorbance changes (GIACs) that represent primary responses of gravitropism (Schmidt and Galland, 2000). GIACs (DeltaA(460-665)) of sporangiophores were measured in vivo with a micro-dual wavelength spectrometer at 460 and 665 nm. Sporangiophores that were placed horizontally displayed an instant increase of the GIACs while the return to the vertical position elicited an instant decrease. The GIACs are specific for graviperception, because they were absent in a gravitropism mutant with a defective madJ gene. During parabola flights hypergravity (1.8 g) elicited a decrease of the GIACs, while microgravity (0 +/- 3 x 10 (-2) g) elicited an instant increase. Hypergravity that was generated in a centrifuge (1.5-6.5 g) elicited also a decrease of the GIACs that saturated at about 5 g. The GIACs have a latency of about 20 ms or shorter and are thus the fastest graviresponses ever measured for fungi, protists, and plants. The threshold for eliciting the GIACs is near 3 x 10 (-2) g, which coincides numerically with the threshold for gravitropic bending. In contrast to gravitropic bending, which requires long-term stimulation, GIACs can be elicited by stimuli as short as 20 to 100 ms, leading to an extremely low threshold dose (acceleration x time) of about 3 x 10 (-3) g s, a value, which is four orders of magnitude below the ones described for other organisms and which makes the GIACs of Phycomyces blakesleeanus the most sensitive gravi-response in literature. PMID- 15122027 TI - Stress tolerance and glucose insensitive phenotypes in Arabidopsis overexpressing the CpMYB10 transcription factor gene. AB - The resurrection plant Craterostigma plantagineum has the ability to survive complete dehydration. In an attempt to further understand desiccation tolerance in this plant, the CpMYB10 transcription factor gene was functionally characterized. CpMYB10 is rapidly induced by dehydration and abscisic acid (ABA) treatments in leaves and roots, but no expression was detected in fully hydrated tissues. Electrophoretic mobility shift assay experiments showed binding of rCpMYB10 to specific mybRE elements within the LEA Cp11-24 and CpMYB10 promoters. Localization of CpMYB10 transcript by in situ reverse transcription-PCR reactions showed expression in vascular tissues, parenchyma, and epidermis both in leaves and roots in response to ABA. Transgenic Arabidopsis plants transformed with CpMYB10 promoter fused to GUS gene showed reporter expression under ABA and stress conditions in several organs. Overexpression of CpMYB10 cDNA in Arabidopsis led to desiccation and salt tolerance of transgenics lines. Interestingly, it was found that plants overexpressing CpMYB10 exhibited Glc insensitive and ABA hypersensitive phenotypes. Therefore, our results indicate that CpMYB10 in Arabidopsis is mediating stress tolerance and altering ABA and Glc signaling responses. PMID- 15122029 TI - Quantitative trait loci associated with drought tolerance at reproductive stage in rice. AB - Drought is a major constraint to rice (Oryza sativa) yield and its stability in rainfed and poorly irrigated environments. Identifying genomic regions influencing the response of yield and its components to water deficits will aid in our understanding of the genetics of drought tolerance and development of more drought tolerant cultivars. Quantitative trait loci (QTL) for grain yield and its components and other agronomic traits were identified using a subset of 154 doubled haploid lines derived from a cross between two rice cultivars, CT9993-510 to 1-M and IR62266-42 to 6-2. Drought stress treatments were managed by use of a line source sprinkler irrigation system, which provided a linearly decreasing level of irrigation coinciding with the sensitive reproductive growth stages. The research was conducted at the Ubon Rice Research Center, Ubon, Thailand. A total of 77 QTL were identified for grain yield and its components under varying levels of water stress. Out of the total of 77 QTL, the number of QTL per trait were: 7 grain yield (GY); 8-biological yield (BY); 6-harvest index (HI); 5-d to flowering after initiation of irrigation gradient (DFAIG); 10-total spikelet number (TSN); 7-percent spikelet sterility (PSS); 23-panicle number (PN); and 11-plant height (PH). The phenotypic variation explained by individual QTL ranged from 7.5% to 55.7%. Under well-watered conditions, we observed a high genetic association for BY, HI, DFAIG, PSS, TSN, PH, and GY. However, only BY and HI were found to be significantly associated with GY under drought treatments. QTL flanked by markers RG104 to RM231, EMP2_2 to RM127, and G2132 to RZ598 on chromosomes 3, 4, and 8 were associated with GY, HI, DFAIG, BY, PSS, and PN under drought treatments. The aggregate effects of these QTL on chromosomes 3, 4, and 8 resulted in higher grain yield. These QTL will be useful for rainfed rice improvement, and will also contribute to our understanding of the genetic control of GY under drought conditions at the sensitive reproductive stage. Close linkage or pleiotropy may be responsible for the coincidence of QTL detected in this experiment. Digenic interactions between QTL main effects for GY, BY, HI, and PSS were observed under irrigation treatments. Most (but not all) DH lines have the same response in measure of productivity when the intensity of water deficit was increased, but no QTL by irrigation treatment interaction was detected. The identification of genomic regions associated with GY and its components under drought stress will be useful for marker-based approaches to improve GY and its stability for farmers in drought-prone rice environments. PMID- 15122028 TI - Interactions between auxin transport and the actin cytoskeleton in developmental polarity of Fucus distichus embryos in response to light and gravity. AB - Land plants orient their growth relative to light and gravity through complex mechanisms that require auxin redistribution. Embryos of brown algae use similar environmental stimuli to orient their developmental polarity. These studies of the brown algae Fucus distichus examined whether auxin and auxin transport are also required during polarization in early embryos and to orient growth in already developed tissues. These embryos polarize with the gravity vector in the absence of a light cue. The auxin, indole-3-acetic acid (IAA), and auxin efflux inhibitors, such as naphthylphthalamic acid (NPA), reduced environmental polarization in response to gravity and light vectors. Young rhizoids are negatively phototropic, and NPA also inhibits rhizoid phototropism. The effect of IAA and NPA on gravity and photopolarization is maximal within 2.5 to 4.5 h after fertilization (AF). Over the first 6 h AF, auxin transport is relatively constant, suggesting that developmentally controlled sensitivity to auxin determines the narrow window during which NPA and IAA reduce environmental polarization. Actin patches were formed during the first hour AF and began to photolocalize within 3 h, coinciding with the time of NPA and IAA action. Treatment with NPA reduced the polar localization of actin patches but not patch formation. Latrunculin B prevented environmental polarization in a time frame that overlaps the formation of actin patches and IAA and NPA action. Latrunculin B also altered auxin transport. Together, these results indicate a role for auxin in the orientation of developmental polarity and suggest interactions between the actin cytoskeleton and auxin transport in F. distichus embryos. PMID- 15122030 TI - Developmental physiology of cluster-root carboxylate synthesis and exudation in harsh hakea. Expression of phosphoenolpyruvate carboxylase and the alternative oxidase. AB - Harsh hakea (Hakea prostrata R.Br.) is a member of the Proteaceae family, which is highly represented on the extremely nutrient-impoverished soils in southwest Australia. When phosphorus is limiting, harsh hakea develops proteoid or cluster roots that release carboxylates that mobilize sparingly soluble phosphate in the rhizosphere. To investigate the physiology underlying the synthesis and exudation of carboxylates from cluster roots in Proteaceae, we measured O2 consumption, CO2 release, internal carboxylate concentrations and carboxylate exudation, and the abundance of the enzymes phosphoenolpyruvate carboxylase and alternative oxidase (AOX) over a 3-week time course of cluster-root development. Peak rates of citrate and malate exudation were observed from 12- to 13-d-old cluster roots, preceded by a reduction in cluster-root total protein levels and a reduced rate of O2 consumption. In harsh hakea, phosphoenolpyruvate carboxylase expression was relatively constant in cluster roots, regardless of developmental stage. During cluster-root maturation, however, the expression of AOX protein increased prior to the time when citrate and malate exudation peaked. This increase in AOX protein levels is presumably needed to allow a greater flow of electrons through the mitochondrial electron transport chain in the absence of rapid ATP turnover. Citrate and isocitrate synthesis and accumulation contributed in a major way to the subsequent burst of citrate and malate exudation. Phosphorus accumulated by harsh hakea cluster roots was remobilized during senescence as part of their efficient P cycling strategy for growth on nutrient impoverished soils. PMID- 15122031 TI - Light induces phosphorylation of glucan water dikinase, which precedes starch degradation in turions of the duckweed Spirodela polyrhiza. AB - Degradation of storage starch in turions, survival organs of Spirodela polyrhiza, is induced by light. Starch granules isolated from irradiated (24 h red light) or dark-stored turions were used as an in vitro test system to study initial events of starch degradation. The starch-associated pool of glucan water dikinase (GWD) was investigated by two-dimensional gel electrophoresis and by western blotting using antibodies raised against GWD. Application of this technique allowed us to detect spots of GWD, which are light induced and absent on immunoblots prepared from dark-adapted plants. These spots, showing increased signal intensity following incubation of the starch granules with ATP, became labeled by randomized [betagamma-33P]ATP but not by [gamma-33P]ATP and were removed by acid phosphatase treatment. This strongly suggests that they represent a phosphorylated form(s) of GWD. The same light signal that induces starch degradation was thus demonstrated for the first time to induce autophosphorylation of starch-associated GWD. The in vitro assay system has been used to study further effects of the light signal that induces autophosphorylation of GWD and starch degradation. In comparison with starch granules from dark-adapted plants, those from irradiated plants showed increase in (1) binding capacity of GWD by ATP treatment decreased after phosphatase treatment; (2) incorporation of the beta-phosphate group of ATP into starch granules; and (3) rate of degradation of isolated granules by starch-associated proteins, further enhanced by phosphorylation of starch. The presented results provide evidence that autophosphorylation of GWD precedes the initiation of starch degradation under physiological conditions. PMID- 15122032 TI - CINCINNATA controls both cell differentiation and growth in petal lobes and leaves of Antirrhinum. AB - To understand how differentiation and growth may be coordinated during development, we have studied the action of the CINCINNATA (CIN) gene of Antirrhinum. We show that in addition to affecting leaf lamina growth, CIN affects epidermal cell differentiation and growth of petal lobes. Strong alleles of cin give smaller petal lobes with flat instead of conical cells, correlating with lobe-specific expression of CIN in the wild type. Moreover, conical cells at the leaf margins are replaced by flatter cells, indicating that CIN has a role in cell differentiation of leaves as well as petals. A weak semidominant cin allele affects cell types mainly in the petal but does not affect leaf development, indicating these two effects can be separated. Expression of CIN correlates with expression of cell division markers, suggesting that CIN may influence petal growth, directly or indirectly, through effects on cell proliferation. For both leaves and petals, CIN affects growth and differentiation of the more distal and broadly extended domains (leaf lamina and petal lobe). However, while CIN promotes growth in petals, it promotes growth arrest in leaves, possibly because of different patterns of growth control in these systems. PMID- 15122033 TI - Phosphatase under-producer mutants have altered phosphorus relations. AB - Phosphorus (P) acquisition and partitioning are essential for plant homeostasis. P is available for plant uptake when in its inorganic form (H2PO4-, or Pi), but Pi is often limiting in soils. Plants secrete acid phosphatases (APases) into the apoplastic space, which may be important for obtaining Pi from organic P sources; however, the relative importance of these enzymes for plant P nutrition has yet to be determined. We demonstrate that the root-associated APase pool is increased in Arabidopsis when Pi is limiting and document five APase isoforms secreted from Arabidopsis roots. Previously, we presented the identification of the phosphatase under-producer (pup) mutants, which have decreased in vivo root APase staining when grown under low P conditions. Here, we present the characterization of one of these, pup3, and further studies with pup1. pup3 has 49%, 38%, and 37% less specific APase activity in exudates, roots, and shoots, respectively. Root associated APase activity is decreased by 16% in pup1 and 25% in pup3, regardless of P treatment. Two APase activity isoforms are reduced in pup3 exudates, and root and shoot isoforms are also affected. One of the two exudate isoforms is recognized by a polyclonal antibody raised to an Arabidopsis purple APase recombinant protein (AtPAP12); however, AtPAP12 transcript levels are unaffected in the mutant. The pup3 mutation was mapped to 68.4 +/- 6.0 centimorgans on chromosome 5. Although P concentrations were not altered in pup1 and pup3 tissues when grown in nutrient solution in which Pi was the sole source of P, the mutants had 10% (pup1) and 17% (pup3) lower shoot P concentrations when grown in a peat vermiculite mix in which the majority of the total P was present as organic P. Therefore, the pup defects, which include secreted APases, are functionally important for plant P nutrition. PMID- 15122034 TI - Two new alleles of the abscisic aldehyde oxidase 3 gene reveal its role in abscisic acid biosynthesis in seeds. AB - The abscisic aldehyde oxidase 3 (AAO3) gene product of Arabidopsis catalyzes the final step in abscisic acid (ABA) biosynthesis. An aao3-1 mutant in a Landsberg erecta genetic background exhibited a wilty phenotype in rosette leaves, whereas seed dormancy was not affected (Seo et al., 2000a). Therefore, it was speculated that a different aldehyde oxidase would be the major contributor to ABA biosynthesis in seeds (Seo et al., 2000a). Through a screening based on germination under high-salt concentration, we isolated two mutants in a Columbia genetic background, initially named sre2-1 and sre2-2 (for salt resistant). Complementation tests with different ABA-deficient mutants indicated that sre2-1 and sre2-2 mutants were allelic to aao3-1, and therefore they were renamed as aao3-2 and aao3-3, respectively. Indeed, molecular characterization of the aao3-2 mutant revealed a T-DNA insertional mutation that abolished the transcription of AAO3 gene, while sequence analysis of AAO3 in aao3-3 mutant revealed a deletion of three nucleotides and several missense mutations. Physiological characterization of aao3-2 and aao3-3 mutants revealed a wilty phenotype and osmotolerance in germination assays. In contrast to aao3-1, both aao3-2 and aao3 3 mutants showed a reduced dormancy. Accordingly, ABA levels were reduced in dry seeds and rosette leaves of both aao3-2 and aao3-3. Taken together, these results indicate that AAO3 gene product plays a major role in seed ABA biosynthesis. PMID- 15122035 TI - Evidence for apoplasmic phloem unloading in developing apple fruit. AB - The phloem unloading pathway remains unclear in fleshy fruits accumulating a high level of soluble sugars. A structural investigation in apple fruit (Malus domestica Borkh. cv Golden Delicious) showed that the sieve element-companion cell complex of the sepal bundles feeding the fruit flesh is symplasmically isolated over fruit development. 14C-autoradiography indicated that the phloem of the sepal bundles was functional for unloading. Confocal laser scanning microscopy imaging of carboxyfluorescein unloading showed that the dye remained confined to the phloem strands of the sepal bundles from the basal to the apical region of the fruit. A 52-kD putative monosaccharide transporter was immunolocalized predominantly in the plasma membrane of both the sieve elements and parenchyma cells and its amount increased during fruit development. A 90-kD plasma membrane H(+)-ATPase was also localized in the plasma membrane of the sieve element-companion cell complex. Studies of [14C]sorbitol unloading suggested that an energy-driven monosaccharide transporter may be functional in phloem unloading. These data provide clear evidence for an apoplasmic phloem unloading pathway in apple fruit and give information on the structural and molecular features involved in this process. PMID- 15122036 TI - Heterogeneous pollen in Chlorophytum comosum, a species with a unique mode of plastid inheritance intermediate between the maternal and biparental modes. AB - The majority of angiosperms display maternal plastid inheritance. The cytological mechanisms of this mode of inheritance have been well studied, but little is known about its genetic relationship to biparental inheritance. The angiosperm Chlorophytum comosum is unusual in that different pollen grains show traits of different modes of plastid inheritance. About 50% of these pollen grains exhibit the potential for biparental plastid inheritance, whereas the rest exhibit maternal plastid inheritance. There is no morphological difference between these two types of pollen. Pollen grains from different individuals of C. comosum all exhibited this variability. Closer examination revealed that plastid polarization occurs, with plastids being excluded from the generative cell during the first pollen mitosis. However, the exclusion is incomplete in 50% of the pollen grains, and the few plastids distributed to the generative cells divide actively after mitosis. Immunoelectron microscopy using an anti-DNA antibody demonstrated that the plastids contain a large amount of DNA. As there is a considerable discrepancy between the exclusion and duplication of plastids, resulting in plastids with opposite fates occurring simultaneously in C. comosum, we propose that the species is a transitional type with a mode of plastid inheritance that is genetically intermediate between the maternal and biparental modes. PMID- 15122038 TI - Analysis of natural allelic variation of Arabidopsis seed germination and seed longevity traits between the accessions Landsberg erecta and Shakdara, using a new recombinant inbred line population. AB - Quantitative trait loci (QTL) mapping was used to identify loci controlling various aspects of seed longevity during storage and germination. Similar locations for QTLs controlling different traits might be an indication for a common genetic control of such traits. For this analysis we used a new recombinant inbred line population derived from a cross between the accessions Landsberg erecta (Ler) and Shakdara (Sha). A set of 114 F9 recombinant inbred lines was genotyped with 65 polymerase chain reaction-based markers and the phenotypic marker erecta. The traits analyzed were dormancy, speed of germination, seed sugar content, seed germination after a controlled deterioration test, hydrogen peroxide (H2O2) treatment, and on abscisic acid. Furthermore, the effects of heat stress, salt (NaCl) stress, osmotic (mannitol) stress, and natural aging were analyzed. For all traits one or more QTLs were identified, with some QTLs for different traits colocating. The relevance of colocation for mechanisms underlying the various traits is discussed. PMID- 15122037 TI - Both subunits of ADP-glucose pyrophosphorylase are regulatory. AB - The allosteric enzyme ADP-Glc pyrophosphorylase (AGPase) catalyzes the synthesis of ADP-Glc, a rate-limiting step in starch synthesis. Plant AGPases are heterotetramers, most of which are activated by 3-phosphoglyceric acid (3-PGA) and inhibited by phosphate. The objectives of these studies were to test a hypothesis concerning the relative roles of the two subunits and to identify regions in the subunits important in allosteric regulation. We exploited an Escherichia coli expression system and mosaic AGPases composed of potato (Solanum tuberosum) tuber and maize (Zea mays) endosperm subunit fragments to pursue this objective. Whereas potato and maize subunits have long been separated by speciation and evolution, they are sufficiently similar to form active mosaic enzymes. Potato tuber and maize endosperm AGPases exhibit radically different allosteric properties. Hence, comparing the kinetic properties of the mosaics to those of the maize endosperm and potato tuber AGPases has enabled us to identify regions important in regulation. The data herein conclusively show that both subunits are involved in the allosteric regulation of AGPase. Alterations in the small subunit condition drastically different allosteric properties. In addition, extent of 3-PGA activation and extent of 3-PGA affinity were found to be separate entities, mapping to different regions in both subunits. PMID- 15122039 TI - Quantitative trait locus analysis of growth-related traits in a new Arabidopsis recombinant inbred population. AB - Arabidopsis natural variation was used to analyze the genetics of plant growth rate. Screening of 22 accessions revealed a large variation for seed weight, plant dry weight and relative growth rate but not for water content. A positive correlation was observed between seed weight and plant area 10 d after planting, suggesting that seed weight affects plant growth during early phases of development. During later stages of plant growth this correlation was not significant, indicating that other factors determine growth rate during this phase. Quantitative trait locus (QTL) analysis, using 114 (F9 generation) recombinant inbred lines derived from the cross between Landsberg erecta (Ler, from Poland) and Shakdara (Sha, from Tadjikistan), revealed QTLs for seed weight, plant area, dry weight, relative growth rate, chlorophyll fluorescence, flowering time, and flowering-related traits. Growth traits (plant area, dry weight, and relative growth rate) colocated at five genomic regions. At the bottom of chromosome 5, colocation was found of QTLs for leaf area, leaf initiation speed, specific leaf area, and chlorophyll fluorescence but not for dry weight, indicating that this locus might be involved in leaf development. No consistent relation between growth traits and flowering time was observed despite some colocations. Some of the QTLs detected for flowering time overlapped with loci detected in other recombinant inbred line populations, but also new loci were identified. This study shows that Arabidopsis can successfully be used to study the genetic basis of complex traits like plant growth rate. PMID- 15122040 TI - Impact of altered gibberellin metabolism on biomass accumulation, lignin biosynthesis, and photosynthesis in transgenic tobacco plants. AB - Gibberellins (GAs) are involved in regulation of many aspects during plant development. To investigate the impact of altered GA levels on plant growth and metabolism, transgenic tobacco (Nicotiana tabacum) plants have been engineered to express either a GA20-oxidase (AtGA20-ox) or a GA2-oxidase (AtGA2-ox) gene from Arabidopsis under control of the cauliflower mosaic virus 35S promoter. Resulting plants were characterized by elongated or stunted shoot growth, respectively, indicating changes in the content of bioactive GAs. In accordance with the effect on plant growth, biomass production was increased or decreased in AtGA20-ox or AtGA2-ox plants, respectively, and was found to be positively correlated with the rate of photosynthesis as determined at the whole plant level. Differences in dry matter accumulation were most likely due to changes in lignin deposition as indicated by histochemical staining and quantitative measurements. Altered lignification of transgenic plants was paralleled by up- or down-regulation of the expression of lignin biosynthetic genes. Short-term GA3 feeding of excised petioles induced lignin formation in the absence of a transcriptional activation of pathway-specific genes. Thus, short-term GA treatment mediates lignin deposition most likely by polymerization of preformed monomers, whereas long-term effects on lignification involve elevated production of precursors by transcriptional stimulation of the biosynthetic pathway. Interestingly, analysis of stem cross sections revealed a differential effect of GA on the formation of xylem and pith cells. The number of lignified vessels was increased in AtGA20-ox plants pointing to a stimulation of xylem formation while the number of pith cells declined indicating a negative regulation. PMID- 15122041 TI - The key role of phloroglucinol O-methyltransferase in the biosynthesis of Rosa chinensis volatile 1,3,5-trimethoxybenzene. AB - 1,3,5-Trimethoxybenzene is a key component of the Chinese rose odor. This compound is synthesized in three successive methylation steps from phloroglucinol, the initial precursor. A novel, to our knowledge, phloroglucinol O-methyltransferase (POMT) characterized here methylates the first step to produce the intermediate 3,5-dihydroxyanisole, while two previously described orcinol O-methyltransferases catalyze the subsequent steps. We isolated POMT from rose petals and determined partial amino acid sequences of the purified enzyme. The full-length POMT cDNA was isolated and expressed in Escherichia coli. Both the native and recombinant POMT exhibited substrate specificity for phloroglucinol. POMT was expressed specifically in floral organs, in accordance with its role as a key enzyme in the synthesis of rose floral scent compounds. PMID- 15122042 TI - Identification of CED-3 substrates by a yeast-based screening method. AB - Identifying cellular substrates repertoire of individual proteases will facilitate our understanding of their physiological and pathological roles. In this article, we employed a yeast-based screening method to isolate CED-3 substrates. This method uses a transcription factor anchored to the plasma membrane by fusion to a library of cellular protein sequences. When a fusion protein is cleaved by CED-3, the transcription factor is released from the plasma membrane and enters the nucleus where it turns on the expression of reporter genes. We identified seven candidate clones by screening a genomic library using this method. Of these seven clones, two were cleaved by purified CED-3 in vitro. Therefore, the method described here may be generally used for genomewide screening to isolate potential substrates of specific proteases. PMID- 15122043 TI - Separate control of Rep and Cap expression using mutant and wild-type loxP sequences and improved packaging system for adeno-associated virus vector production. AB - Adeno-associated virus (AAV) vectors are a practical choice for gene transfer, and demand for them is increasing. To cope with the necessity in the near future, we have developed a number of approaches to establish packaging cell lines for the production of AAV vectors. In our previous study, a highly regulated expression of large Rep proteins was obtained by using the Cre-loxP switching system. Therefore, in the present study, to regulate Cap expression as well, we developed an inducible expression system for both Rep and Cap proteins by using an additional set of mutant loxP sequences. The mutants possess two base alterations in the spacer region of loxP and recombine specifically with the same counterpart in the presence of Cre. By using two separate plasmids, one with mutant and the other with wild-type loxP sequences, the expression of two different proteins can be induced simultaneously by Cre recombinase. When the LacZ-encoding plasmid vector was used as a packaging model, a significant packaging titer of 2.1 x 1010 genome copies per 10-cm dish was obtained. These results indicate the importance of controlling Cap expression, in addition to Rep, to achieve an optimum production rate for AAV vectors. PMID- 15122044 TI - Identification of the role of a cysteine-rich region of PC6B by determining the enzymatic characteristics of its mutants. AB - In the proprotein convertases family, mouse PC6B (mPC6B) has a very large cysteine-rich region (CRR), consisting of 22 tandem cysteine-rich (Cys-rich) repeated segments. The role of this region remains elusive. In this report, to get insight on the possible role of the CRR, we constructed four truncated mPC6B mutant genes with 0, 5, 11, and 22 Cys-rich repeated segments remaining; using the baculovirus-expression system and a simple purification method, we obtained four enzyme mutants of mPC6B. By determining their optimal pH and calcium ion concentration for enzymatic activity and their thermal stability, we found that CRR did not affect pH optimum and Ca2+ optimum compared with the p-domain. However, CRR acted as a stabilizing domain in addition to the p-domain. By kinetic analyses of four mutants, we found that the long Cys-rich repeats in the native form of mPC6B reduced its Vmax. These facts suggest that CRR acts as an important part of functional domain. PMID- 15122045 TI - Rat kidney-targeted naked plasmid DNA transfer by retrograde injection into the renal vein. AB - Kidney-targeted gene transfer is expected to revolutionize the treatment of renal diseases. Recently, we demonstrated that naked plasmid deoxyribonucleic acid (DNA) can be transferred into renal interstitial fibroblasts near the peritubular capillaries (PTCs) in normal rats, by retrograde injection into the renal vein with the renal vein and artery clamped. The PTC network is a main target of kidney transplant rejection and of progressive tubulointerstitial fibrosis, which typifies all progressive renal diseases. We retrogradely injected a lacZ expression plasmid in Ringer's solution into the renal vein of rats using a 24 gage catheter. We detected lacZ expression exclusively in the interstitial fibroblasts near the PTCs of the kidney by immunoelectron microscopy. Nephrotoxicity from the gene transfer was not apparent. We then used a rat erythropoietin (Epo) expression plasmid vector pCAGGS-Epo in a reporter assay. We obtained maximal Epo expression when the DNA solution was injected within 5 s in a volume of 1.0 mL. We detected transgene-derived Epo messenger ribonucleic acid by reverse transcriptase polymerase chain reaction only in the kidneys receiving pCAGGS-Epo. In this article, protocols for naked plasmid DNA transfer into rat kidney using this hydrodynamics-based transfection method and the immunoelectron microscopic technique to determine the lacZ gene transfer site are described in detail. PMID- 15122048 TI - SNP discrimination through proofreading and OFF-switch of exo+ polymerase. AB - Single nucleotide polymorphisms (SNPs) are useful physical markers for genetic studies as well as the cause of some genetic diseases. To develop more reliable SNP assays, we examined the underlying molecular mechanisms by which deoxyribonucleic acid (DNA) polymerases with 3' exonuclease activity maintain the high fidelity of DNA replication. In addition to mismatch removal by proofreading, we have discovered a premature termination of polymerization mediated by a novel OFF-switch mechanism. Two SNP assays were developed, one based on proofreading using 3' end-labeled primer extension and the other based on the newly identified OFF-switch, respectively. These two new assays are well suited for conventional techniques, such as electrophoresis and microplates detection systems as well as the sophisticated microchips. Application of these reliable SNP assays will greatly facilitate genetic and biomedical studies in the postgenome era. PMID- 15122046 TI - Calmodulin's flexibility allows for promiscuity in its interactions with target proteins and peptides. AB - The small bilobal calcium regulatory protein calmodulin (CaM) activates numerous target enzymes in response to transient changes in intracellular calcium concentrations. Binding of calcium to the two helix-loop-helix calcium-binding motifs in each of the globular domains induces conformational changes that expose a methionine-rich hydrophobic patch on the surface of each domain of the protein, which it uses to bind to peptide sequences in its target enzymes. Although these CaM-binding domains typically have little sequence identity, the positions of several bulky hydrophobic residues are often conserved, allowing for classification of CaM-binding domains into recognition motifs, such as the 1-14 and 1-10 motifs. For calcium-independent binding of CaM, a third motif known as the IQ motif is also common. Many CaM-peptide complexes have globular conformations, where CaM's central linker connecting the two domains unwinds, allowing the protein to wrap around a single predominantly alpha-helical target peptide sequence. However, novel structures have recently been reported where the conformation of CaM is highly dissimilar to these globular complexes, in some instances with less than a full compliment of bound calcium ions, as well as novel stoichiometries. Furthermore, many divergent CaM isoforms from yeast and plant species have been discovered with unique calcium-binding and enzymatic activation characteristics compared to the single CaM isoform found in mammals. PMID- 15122047 TI - Preclinical safety testing of biotechnology-derived pharmaceuticals: understanding the issues and addressing the challenges. AB - The unique and complex nature of biotechnology-derived pharmaceuticals has meant that it is often not possible to follow the conventional safety testing programs used for chemicals, and hence they are evaluated on a case-by-case basis. Nonclinical safety testing programs must be rationally designed with a strong scientific understanding of the product, including its method of manufacture, purity, sequence, structure, species specificity, pharmacological and immunological effects, and intended clinical use. This knowledge, coupled with a firm understanding of the regulatory requirements for particular product types, will ensure that the most sensitive and regulatory-compliant test systems are used to optimize the chances of gaining regulatory approval for clinical testing or marketing authorization in the shortest possible time frame. PMID- 15122049 TI - Comparison of the immunogenicity and safety of two different brands of Salmonella typhi Vi capsular polysaccharide vaccine. AB - BACKGROUND: The recent emergence of multi-drug-resistant Salmonella strains highlights the need for better preventive measures, including vaccination. Safe and immunologic vaccines have been developed based on purified Vi polysaccharide. OBJECTIVE: To compare the immune response elicited by two different brands of Salmonella Vi capsular polysaccharide vaccine (ViCPS). SETTING AND DESIGN: Double blind, randomized (3:1), controlled, parallel, phase III study was conducted at two centres in India to compare the safety and immunogenicity of Typbar, the investigational vaccine with an already marketed vaccine "X", in healthy subjects aged between 12 -25 years. MATERIAL AND METHODS: A sample size of 184 subjects was calculated. Subjects were randomly distributed in two groups, immunized with single dose of Typbar or Vaccine "X". Serum samples were taken before 7 days and 4 weeks after immunization for the determination of antibodies to Vi polysaccharide, by ELISA method. Safety was assessed by physical examination, laboratory parameters before and after vaccination and by monitoring adverse events. STATISTICS: The geometric mean antibody titre (GMT) 4 weeks after vaccination was compared from respective pre-vaccination values by Wilcoxon signed rank test. Geometric mean of antibody levels before and after immunization and the ratio between them (Mann-Whitney test), the Seroconversion rates (Z test of proportions) and the adverse events (Fisher's exact test and Chi square test), were compared between two groups. P value < 0.05 was considered statistically significant. P values and 95% confidence intervals were estimated in two-tailed fashion. RESULTS: 153 subjects (Typbar =116 and Vaccine "X" =37) were studied. 71.6% (95% CI=63.4%-79.8%) and 75.7% (95% CI=64.9% - 89.5%) were the seroconversion rates with Typbar and vaccine "X" respectively. The GMT values for Vi antibodies induced after Typbar and vaccine "X" were 10.23 Typbar and 13.46 mg/mL respectively and these values showed high significance when compared to their respective pre-immunization GMT values (P<0.0001) at 95% CI (-10.49 to 7.19 mg/mL for Typbar and -14.69 to -8.86 mg/mL for Vaccine "X"). The induction of antibody response appeared to be slightly stronger (P=0.032) with vaccine "X" when compared to that of Typbar. This is justifiable as the same group also had high pre-immunization GMT values (P=0.021). CONCLUSION: The immunogenicity and safety of the investigational vaccine Typbar was found to be similar to that of already marketed brand of Vi CPS, Vaccine "X". The availability of a single dose of vaccine that is safe and effective enhances the prospective for control of typhoid fever. PMID- 15122050 TI - Effect of estrogen on plasma ceruloplasmin level in rats exposed to acute stress. AB - BACKGROUND: Plasma ceruloplasmin, a copper containing protein, belongs to a class called acute phase proteins. Reduced level of ceruloplasmin was associated with Wilson's disease and Menke's kinky hair disease in man, primarily affecting copper metabolism. Stress was known to increase Ceruloplasmin. Several stress associated changes were commonly observed in women at menopause and also those who underwent overiectomy. Present experiment investigated the effect of estrogen on ceruloplasmin level in acute stress. AIMS: To assess the estradiol induced changes in plasma ceruloplasmin concentration on exposure of the rats to acute stress. SETTINGS AND DESIGN: Acute stress was induced by forcing the rats to swim till exhaustion. The rats were overiectomised bilaterally to remove the primary source of sex hormones. And hormone replacement was done later. MATERIAL AND METHODS: Wistar albino female rats were used. Acute stress was induced before overiectomy, following recovery from surgery, and again after Estradiol Valerate injection (for 10 days) in same group of rats. The plasma ceruloplasmin was estimated immediately after stress during each stage--that is preoperative control, stressed control, after overiectomy and then following treatment with Estradiol Valerate. STATISTICAL ANALYSIS USED: Paired sample T test was applied to analyze the findings. RESULTS: We found lowest ceruloplasmin level after stress in overiectomised animals, while on substitution of estradiol the trend appeared to be reversed. CONCLUSION: The result suggested a direct effect of estrogen on hepatic ceruloplasmin production/release and this could account for some of the beneficial effects of hormone replacement therapy. PMID- 15122051 TI - Effect of undernutrition and subsequent rehabilitation on brain 5-HT (serotonin) profile of developing newborn rats. AB - For littermate comparison, undernutrition in half of the pups of a litter was induced by 12h maternal deprivation from day 5 to day 18 postnatal. Subsequently, undernourished suckling rats were rehabilitated. Brain 5-HT concentrations and its turnover rate were measured on different days of age up to the period of 3 months. Brain 5-HT concentration was approximately half of the adult level at birth and increased progressively, except with a steep fall on day 18 postnatal, to the adult level by day 25 postnatal and thereafter it did not vary much on different days of age. Undernourished newborn rats showed significantly low level of brain 5-HT on day 9 and significantly higher concentrations on day 12 and 18 postnatal, whereas, rehabilitated rats showed comparable brain 5-HT concentrations with that of their nourished littermates. It is concluded that undernutrition had affected the brain 5-HT concentration and its turnover rate. Brain 5-HT concentration was low during early phase and was more during late phase of undernutrition. Rehabilitation of undernourished rats restored the brain 5-HT concentration similar to that of their nourished littermates. PMID- 15122052 TI - Salmonella enterica serotype paratyphi:A induced acute diarrhea. PMID- 15122053 TI - Prevalence of overweight and obesity amongst elderly patients attending a geriatric clinic in a tertiary care hospital in Delhi, India. PMID- 15122054 TI - Effect of Udad Dal and iron syrup on growth and hemoglobin level in rural school children. PMID- 15122055 TI - A new method of color Doppler perfusion measurement via dynamic sonographic signal quantification in renal parenchyma. AB - OBJECTIVE: Perfusion quantification of tissues is an important goal to evaluate the state of blood supply of an organ. We developed a method to quantify tissue perfusion via color Doppler signal quantification from sonographic videos and applied this to describe renal parenchymal perfusion in healthy kidneys. METHOD: Color Doppler sonographic videos of renal perfusion from both kidneys of 87 healthy children (age 2 weeks to 16 years) were recorded under defined conditions. Perfusion data (color hue, color area) were measured in a standardized region of interest automatically. Signal intensity was calculated as whole ROIs (regions of interest) mean flow velocity (cm/s) encoded by color Doppler signals during one full heart cycle. RESULTS: Normal signal intensity values are: 1.86 cm/s in the region encompassing central 50% of the renal cortex and 0.56 cm/s in the peripheral 50% of the renal cortex. These differences are significant. Signal intensity of both kidneys did not differ. CONCLUSION: Signal intensity of cortical tissue in healthy kidneys was quantified noninvasively from color Doppler signal data in an easily accomplishable manner with new measurement software. Normal values for this technique have been calculated. Possible further applications might be all situations, where perfusion changes could be expected like inflammation, renal insufficiency, vascular diseases and tumors. PMID- 15122056 TI - Oxidative stress influences CC-chemokine levels in hemodialyzed patients. AB - BACKGROUND: Increased oxidative stress (SOX) has been reported in hemodialyzed (HD) patients, but its influence on CC-chemokine levels remains unknown. METHODS: The levels of 3 distinct SOX markers (Cu/Zn superoxide dismutase (Cu/Zn SOD), total lipid peroxide, and autoantibodies against oxidized LDL (OxLDL-Ab)), as well as those of 4 CC-chemokines (monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory proteins (MIP-1alpha, MIP-1beta), regulated upon activation, normal T cell expressed and secreted (RANTES), and tumor necrosis factor-alpha (TNF-alpha)) were measured pre- and post-HD session in 15 HD patients with cardiovascular disease (CVD) and 12 HD patients without CVD (pre- and post-HD session) and 17 controls. RESULTS: Cu/Zn SOD levels were elevated in HD patients (both before and after HD) compared to controls (p < 0.001). Total lipid peroxide levels were similar in the controls in pre-dialysis samples, but were increased after a HD session (p < 0.001). Pre-dialysis OxLDL-Ab levels were increased only in the HD group with CVD compared to controls (p < 0.05). The pre dialysis plasma levels of TNF-alpha (p < 0.001), MCP-1 (p < 0.001) and MIP-1beta (p < 0.01) were increased both in all HD patients and subgroups with CVD when compared to the controls, and remained significantly elevated when measured after HD. Dialysis increased MCP-1 (p < 0.05) and MIP-1beta (p < 0.001) levels as compared to the controls as well as in patients without CVD. Plasma RANTES was significantly lower before HD (p < 0.05) and after HD (p < 0.001) in patients with CVD. Patients without CVD and all HD patients also had lower RANTES before and after HD when compared to controls (all p < 0.001). A positive correlation was observed between plasma pre-dialysis Cu/Zn SOD levels and the other SOX markers (all p < 0.05), age (p < 0.05) and duration of hemodialysis (p < 0.001). A positive relationship existed between plasma Cu/Zn SOD levels and those of MCP 1, MIP-1beta and TNF-alpha (all p < 0.001). RANTES levels negatively correlated with Cu/Zn SOD (p < 0.0001).TNF-alpha positively correlated with age (p < 0.05), total lipid peroxide (p = 0.010), MCP-1 (p < 0.01) and MIP-1beta levels (p < 0.01). An identifiable association exists between MCP-1 and MIP-1beta levels (p < 0.01), whereas both MCP-1 and MIP-1beta were inversely correlated with RANTES (both p < 0.05). CONCLUSION: Our data indicate for the first time a probable functional relationship between oxidative stress and CC-chemokine levels in hemodialyzed patients, particularly in those with cardiovascular disease. This relationship may represent one of the mechanisms involved in the progression of atherosclerosis in these patients. PMID- 15122057 TI - Intestinal absorption and biliary secretion of zinc in rats with chronic renal failure. AB - BACKGROUND/AIMS: In chronic renal failure (CRF), zinc deficiency is partially attributed to decreased intestinal zinc absorption, but the mechanism of this decrease in intestinal zinc absorption is obscure. Therefore, the objective of this study was to investigate the cause of decreased intestinal zinc absorption in a uremic rat model using an in vivo perfusion technique as well as to evaluate the effect of intestinal zinc perfusion on the secretion of biliary zinc in normal and CRF rats. METHODS: CRF was induced by five-sixths nephrectomy (Nx). During zinc sulfate perfusion, absorption of zinc in the small intestine and the response of plasma zinc level were measured. After intestinal zinc perfusion for 80 min, the concentrations of zinc and metallothionein (MT) in the intestinal mucosal and liver tissue were assayed. The secretion of biliary zinc and the excretion of urinary zinc were also determined before and after zinc sulfate perfusion. The results were compared with those obtained from 10 sham-operated normal rats. RESULTS: The CRF rats showed a significant decrease in the rate of intestinal zinc absorption and in the response of plasma zinc levels during intestinal zinc perfusion. They also had significantly higher levels of mucosal zinc and MT than sham-operated normal rats, but their contents of liver zinc were significantly lower than those of sham-operated normal rats after zinc sulfate perfusion. CRF rats showed a low plasma zinc level and a high urinary zinc excretion in baseline levels, but had similar output of basal biliary zinc as compared with sham-operated normal rats. Zinc sulfate perfusion in the small intestinal was not found to increase the secretion of biliary zinc and the excretion of urinary zinc, either in normal or CRF rats. CONCLUSION: In the CRF rat, the reduction of intestinal zinc absorption may result from reduced mucosal zinc efflux from the basolateral membrane into plasma. These data also suggest that the absorbed zinc from the gastrointestinal tract is mostly taken up by the liver or other tissues, and is less excreted in bile juice and urine. PMID- 15122058 TI - Human cortical fibroblast responses to high glucose and hypoxia. AB - BACKGROUND/AIMS: Approximately 30% of individuals with diabetes mellitus are susceptible to diabetic nephropathy, whereas ischemic injury uniformly induces renal impairment. As matrix accumulation correlates with progressive renal disease we assessed parameters associated with matrix turnover in response to high glucose +/- hypoxia in human cortical fibroblasts (CF). METHODS: CF were grown to confluence and exposed to media containing 5 or 25 mmol/l D-glucose for 72 h with or without a superimposed hypoxic insult. RESULTS: High glucose increased cellular protein content (p < 0.05). Combined high glucose and hypoxia induced a further increase in cellular protein content (p < 0.005), suggestive of a synergistic hypertrophic effect. MMP secretion corresponded inversely with changes in TIMP expression. In cell cultures derived from 2/3 of patients, high glucose increased MMP-9 (p < 0.0005) and MMP-2 (p < 0.005) while TIMP-1 was reduced (p = 0.05). In the remaining cell cultures derived from 1/3 of patients, MMP-2 was reduced (p < 0.0001) while TIMP-1 and TIMP-2 were both increased (p < 0.05). In contrast, hypoxia induced uniform reductions in MMP-9 and MMP-2 in both normal and high glucose conditions. High glucose increased the expression of PAI 1 mRNA (p < 0.05) in all patients independent of changes in the MMP-TIMP axis. CONCLUSIONS: In summary, variability was observed in the MMP-TIMP axis following exposure to high glucose. In contrast, high glucose uniformly induces PAI-1 expression. Hypoxic insults uniformly reduce matrix breakdown independent of the prevailing glucose conditions. PMID- 15122059 TI - Effect of dietary creatine on skeletal muscle myosin heavy chain isoform expression in an animal model of uremia. AB - BACKGROUND: Chronic renal failure is accompanied with muscle dysfunction and myopathy, characterized by muscle weakness and increased fatigue. Myosin heavy chain (MHC) is the principal structural protein that controls the intrinsic contractile properties of striated muscle. Creatine is widely used as an ergogenic nutritional supplement in sportsmen. This study investigates the effect of creatine supplementation on MHC expression in the setting of uremic myopathy. METHODS: Male Wistar rats were either sham operated or subtotally nephrectomized and received a control diet or creatine (2% w/w)-supplemented diet. After 4 weeks of treatment, serum creatinine, creatine, urea and creatinine clearances were determined. MHC isoforms were determined electrophoretically in the extensor digitorum longus and soleus muscles. RESULTS: Creatinine clearances were lower in nephrectomized animals, but similar in creatine-supplemented and control diet animals. Nephrectomized animals had significantly higher MHC IIb and lower MHC IIx isoform expression in the extensor digitorum longus muscle compared to sham operated animals. In the soleus muscle, MHC IIb expression was increased in nephrectomized animals. Creatine supplementation reversed the MHC transitions observed in uremia in the soleus muscle, but not in the extensor digitorum longus muscle. CONCLUSION: We observed altered expression of MHC isoforms in uremia. In uremic animals, fast MHC IIb isoforms were increased, whereas MHC I and IIx isoforms predominate in control animals. Dietary creatine supplementation reversed the altered MHC expression during uremia in slow-twitch, but not in fast twitch muscles. PMID- 15122060 TI - Selective expression of TGF-beta2 and TGF-beta3 isoforms in early mesangioproliferative glomerulonephritis. AB - BACKGROUND/AIM: Evidence from ex vivo glomerular analysis has implicated overexpression of transforming growth factor (TGF) beta 1 in progressive renal disease. The roles of TGF-beta2 and TGF-beta3 are less clear. The purpose of this study was to define the temporal expression and abundance of TGF-beta isoforms in both acute and progressive Thy-1 glomerulonephritis during the crucial initiation phase of these models. METHODS: Acute Thy-1 glomerulonephritis was induced by a single injection of OX7, while the progressive model was induced by two injections, 7 days apart. RESULTS: Cellular infiltration of glomeruli consisted of transient increases of neutrophils and ED1+ macrophages. The distribution of TGF-beta1, TGF-beta2, and TGF-beta3 revealed distinct differences in normal and nephritic rats. No changes in TGF-beta1 staining were observed within glomeruli of either model. In marked contrast, in the one-shot model, TGF-beta2 and TGF beta3 stainings increased rapidly, yet transiently, throughout affected glomeruli, followed by more sustained staining in glomerular epithelial cells. Diffuse, transient staining was absent in two-shot glomerulonephritis, but an increase in epithelial cell staining mirrored that seen in the one-shot model. CONCLUSION: Based on these results, we propose that the effects, formerly thought of as solely due to a single entity, TGF-beta1, may be the result of an interplay between individual TGF-beta isoforms. PMID- 15122061 TI - Successful gene transfer using adeno-associated virus vectors into the kidney: comparison among adeno-associated virus serotype 1-5 vectors in vitro and in vivo. AB - BACKGROUND/AIM: Gene transfer into the kidney has great potential as a novel therapeutic approach. However, an efficient method of gene transfer into the kidney has not been established. We explored the transduction efficiency of renal cells in vitro and in vivo using adeno-associated virus (AAV) serotype 1-5 vectors encoding the beta-galactosidase gene. METHODS: In the in vitro study, rat kidney epithelial cell line NRK52E cells were transfected with AAV serotype derived vectors. In the in vivo study, AAV serotype derived vectors were selectively injected into the kidney using a catheter-based gene delivery system in rats and mice mimicking the clinical procedure. The efficiency of gene expression was histologically evaluated on the basis of the beta-galactosidase expression. RESULTS: AAV serotype 1, 2, and 5 vectors transduced in rat kidney epithelial cell line NRK52E cells in vitro, whereas AAV serotype 3 or 4 vectors showed no transduction. In addition, the kidney-specific injection of AAV serotype 2 vectors successfully transduced in tubular epithelial cells, but not in glomerular, blood vessel, or interstitial cells in vivo, whereas the rest of the serotypes showed no transduction. CONCLUSION: Since kidney-specific gene delivery via the renal artery by catheterization is highly feasible in humans, these findings provide useful information for promising strategies in renal gene therapy. PMID- 15122062 TI - Accelerated glomerular injury in hemi-nephrectomized transgenic mice of mesangial cell-predominant serpin, megsin. AB - Mesangial cells play a critical role in the maintenance of normal glomerular functions such as matrix remodeling and immune complex disposal. We recently identified a novel human mesangium-predominant gene, megsin, which is a new member of the serine protease inhibitor (serpin) superfamily. While our previous studies demonstrated progressive mesangial matrix expansion and an increase in the number of mesangial cells in megsin transgenic mice, it took 40 weeks to develop these manifestations. Here we performed hemi-nephrectomy to accelerate glomerular injury in megsin transgenic mice. Hemi-nephrectomized transgenic mice developed focal segmental mesangial expansion, which was associated with proteinuria. Megsin has thus a biologically relevant influence on the development of glomerular damage. The hemi-nephrectomized model of this transgenic mouse might serve as a tool to investigate the mechanisms of glomerular disease. PMID- 15122063 TI - Rheumatoid arthritis and the kidney: uneasy companions. PMID- 15122064 TI - Renal disease as a predictor of increased mortality among patients with rheumatoid arthritis. AB - AIMS AND METHODS: Mortality among RA patients and controls was analyzed with special attention to renal disease in population-based material (originally screened in 1988) of 604 patients with RA (470 females, 134 males) and 457 age- and sex-matched controls (352 females, 105 males). In the original RA population, isolated hematuria (HU) was observed in 54, isolated proteinuria (PU) in 27, combined hematuria and proteinuria (HUPU) in 7, chronic renal failure (CRFtot) in 36 and isolated chronic renal failure without HU or PU (CRFisol) in 15 patients. Among the controls, HU was observed in 39, PU in 11, CRFtot in 32 and CRFisol in 16 subjects. HUPU was not observed in any of the controls. Microalbuminuria (20 200 microg/min) was observed in 34 RA patients and in 27 controls. Histologically confirmed amyloidosis was found in 13 RA patients and mesangial glomerulonephritis (MesGN) in 17 patients. The mortality was evaluated in 1999 from data of the Statistical Office of Finland. Statistical analysis was performed by Cox regression analysis. RESULTS: Mortality was significantly increased in the RA population as compared to controls: hazard ratio (HR) 1.78 (95% CI 1.34-2.31) for all RA patients; HR 2.12 (1.52-2.94) for females; HR 1.15 (0.75-1.77) for males. In the RA material, increased mortality was detected in patients with HUPU (HR 4.45; 1.54-12.84), PU (HR 3.54; 1.88-6.65), CRFtot (HR 3.74; 2.55-5.56) or microalbuminuria (HR 2.77; 1.64-4.69) when compared to those with normal clinical renal findings, whereas HU (HR 1.49; 0.88-2.52), CRFisol (HR 1.71; 0.82-3.54), bacteriuria (HR 0.96; 0.35-2.59) or pyuria (HR 0.65; 0.09-4.65) did not predict mortality. Renal amyloidosis was associated with an over twofold mortality rate (HR 2.31; 1.03-5.15), whereas mortality was within expected limits in RA patients with MesGN (HR 1.61; 0.49-5.24). CONCLUSION: Our results show that nephropathy presenting with combined hematuria and proteinuria, proteinuria, microalbuminuria or histologically confirmed amyloidosis is associated with increased mortality in RA patients, whereas mortality is within expected limits in those with isolated hematuria or mesangial glomerulonephritis. PMID- 15122065 TI - Glomerular capillary tenascin staining in renal allografts and its correlation with capillary loop 'double contours', proteinuria and graft outcome. AB - BACKGROUND: Glomerular capillary injuries result in proteinuria, previously reported to be a risk factor for renal allograft dysfunction. According to the Banff 97 Working Classification of Renal Allograft Pathology, 'double contours' in glomerular capillary loops (GC-DC) were suggested to be most specific for chronic transplant glomerulopathy. Tenascin-C (TN), a component of extracellular matrix, is known to be overexpressed in various conditions. METHODS: TN immunostaining was performed by the labeled streptavidin biotin method. The correlations between glomerular capillary TN (GC-TN) and proteinuria, and between GC-DC and GC-TN, were studied in 27 biopsies showing chronic allograft nephropathy (CAN) lesions. In 14 patients (serum creatinine <2.0 mg/dl at biopsy), graft outcome was studied. Graft function 3 years after biopsy was classified as stable or deteriorated; the term 'deteriorated' was used if serum creatinine had increased 20% over baseline, or if the patient required dialysis. RESULTS: In CAN, GC-TN correlated with GC-DC. Proteinuria in patients with intense GC-TN tended to increase during 1 year after biopsy. In graft outcome analysis, GC-TN was significantly lower in stable grafts than in deteriorated ones. Furthermore, all 3 patients demonstrating weak GC-TN without GC-DC returned to dialysis. CONCLUSION: TN immunostaining is a sensitive method to detect glomerular capillary injury with a predictive value for renal allograft dysfunction. PMID- 15122066 TI - Usefulness of microalbuminuria in cardiovascular risk stratification of essential hypertensive patients. AB - BACKGROUND/AIMS: To evaluate the influence of microalbuminuria (albumin excretion rate--AER) determination and echocardiography (ECHO) on cardiovascular risk stratification, initially performed according the 1999 WHO/ISH guidelines by using only routine diagnostic procedures with or without fundal examination. METHODS: 312 essential hypertensives attending our institution were studied retrospectively. Cardiovascular risk was assessed in a semiquantitative way using four categories of absolute cardiovascular disease risk (low, medium, high and very high risk), as proposed by the 1999 WHO/ISH guidelines, on the basis of data on the average 10-year risk of cardiovascular events among participants in the Framingham Study. RESULTS: Without the retinal data, estimating the level of global cardiovascular risk on the basis of routine work-up alone, 14% were classified as low-risk patients, 48% were as medium-risk, 20% as high-risk and 18% at very-high-risk patients. The combined use of AER and ECHO, in line with the newer ESH-ESC guidelines, determined a statistically significant reclassification of the hypertensive patients. Only 10% remained in the low-risk category, 28% were classified in the medium-, 42% in the high- and 20% in the very-high-risk classes. The overall percentage of patients that changed risk stratum (mostly shifting from the medium- to the high-risk class) was significantly different from the proportion of subjects reclassified after the addition of either microalbuminuria or echocardiography alone. No change in the distribution of risk categories was observed when AER assay and ECHO were added to routine procedures including funduscopic examination. CONCLUSIONS: Considering the questionable prognostic value of qualitative retinal examination, our results suggest that cardiovascular risk evaluation based only on simple routine work-up, ignoring the information provided by AER determination and ECHO, may underestimate the level of absolute risk. PMID- 15122067 TI - Association of FMF-related (MEFV) point mutations with secondary and FMF amyloidosis. AB - BACKGROUND: Familial Mediterranean fever (FMF) is the major cause of AA amyloidosis in Turkey. M694V mutation in MEFV gene was suggested to be associated with severe clinical features and amyloidosis of FMF. METHODS: In this study, the frequencies of three FMF-related MEFV mutations (M694V, M680I and V726A) were investigated in FMF patients with (AA-FMF, n = 37) and without amyloidosis (non AA-FMF, n = 35), in patients with secondary amyloidosis related to non-FMF inflammatory conditions (S-AA, n = 19) and in a non-inflammatory control group (n = 185) by molecular genetic studies using polymerase chain reaction with the ARMS (amplification refractory mutation system) method. RESULTS: Both AA and non-AA FMF patients had significantly higher MEFV mutations compared to non-inflammatory controls (81 and 62.7% respectively vs. 4.2%, p = 0.0001). AA-FMF patients carried significantly more MEFV mutations than non-AA-FMF patients (p = 0.01). M694V was the most common mutation in both FMF groups (63.5 vs. 51.4%), however allele frequency (p = 0.17) and the number of homozygous patients for this mutation did not differ between the groups (p = 0.77). Although lower compared to FMF patients, S-AA patients also had a significantly higher incidence of MEFV mutations than non-inflammatory controls (21 vs. 4.2%) (p = 0.0002). M694V was the only MEFV mutation in this group. CONCLUSION: MEFV mutations are found to be increased both in FMF and non-FMF associated secondary amyloidosis in our study; however, no clear association between M694V and amyloidosis is observed, except in the non-FMF group. Our results suggest that MEVF mutations may also serve as a severity marker for other inflammatory conditions. PMID- 15122068 TI - Hyaluronic-acid butyric esters as promising antineoplastic agents in human lung carcinoma: a preclinical study. AB - New promising compounds, derived from the esterification of hyaluronic acid with butyric acid, were investigated in vitro on a non-small cell lung carcinoma cell line (NCI-H460) and an its metastatic subclone (NCI-H460M). All new compounds exerted a dose-dependent inhibitory effect on both cell lines, which expressed CD44, the specific surface receptor for hyaluronic acid, in a very high percentage of cells (90%). HE1, the most effective of these compounds, was 10 fold more effective than sodium butyrate (NaB) in inhibiting cell proliferation. Similarly to NaB, after 24 hours of treatment, HE1 affected the expression of three cell cycle-related proteins (p27(kip1), p53 and p21(waf1)) responsible for growth arrest, indicating that the presence of the hyaluronic acid backbone does not interfere with the biologic activity. Intratumoral treatment with HE1 demonstrated a marked efficacy on primary tumor growth and on lung metastases formation of the murine Lewis Lung Carcinoma model. Altogether, present findings suggest a possible clinical application of these novel butyric pro-drugs in primary and metastatic lung cancer. PMID- 15122069 TI - Efficacy of novel P-glycoprotein inhibitors to increase the oral uptake of paclitaxel in mice. AB - P-glycoprotein inhibitors can increase the oral bioavailability of paclitaxel. We have now explored the mechanisms that determine the efficacy of several novel P glycoprotein inhibitors to increase the absorption of paclitaxel from the gut lumen of mice in both in vivo and in vitro experiments. The inhibitors studied were cyclosporin A, PSC 833, GF120918, LY335979 and R101933. Mass balance studies showed that GF120918 was the most effective inhibitor, resulting in almost complete uptake of paclitaxel. PSC 833 was slightly less effective, whereas cyclosporin A and LY335979 were moderately effective. R101933 had only marginal effects. These findings were in line with in vitro transport experiments using LLC-mdr1a cells. By studying the intra-intestinal kinetics of the agents we found that cyclosporin A, PSC 833 and GF120918 rapidly passed the stomach and traveled concurrently with paclitaxel through the intestines, whereas LY335979 and R101933 delayed stomach emptying. Moreover, these latter compounds appear to be more readily absorbed when released into the intestines thus reducing local intestinal concentrations. Due to their combined effects on absorption and metabolic elimination of paclitaxel, cyclosporin A and PSC 833 resulted in the highest paclitaxel levels in plasma. In conclusion, our models provide insight into the factors that determine the suitability of P-glycoprotein inhibitors to enable oral paclitaxel therapy and will be useful in selecting candidate inhibitors for clinical testing. PMID- 15122070 TI - Preliminary pharmacokinetic and bioanalytical studies of SJG-136 (NSC 694501), a sequence-selective pyrrolobenzodiazepine dimer DNA-cross-linking agent. AB - SJG-136 is a synthetic pyrrolobenzodiazepine (PBD) dimer in which two DNA alkylating subunits are linked through an inert propanedioxy tether. Biophysical and biochemical studies of SJG-136 have shown a remarkable affinity for DNA and potent cytotoxicity in vitro. On this basis, together with its unique sequence selectivity and interstrand DNA cross-linking activity, SJG-136 has been selected for clinical trials. This study examines the pharmacological characteristics of SJG-136 and provides the first report of pharmacokinetic properties for this agent. A sensitive, selective and reproducible reversed-phase gradient LC/MS assay has been developed for detection and analysis, where a molecular ion ( m / z 557.2) is detectable for the SJG-136 parent imine. Fluorescence detection (260 nm excitation, 420 nm emission) gives a limit of sensitivity of 5 nM (2.5 ng ml( 1)) for analysis of SJG-136 in mouse plasma. Extraction efficiencies from plasma were >65% across a range of concentrations (5-1000 nM). Following administration to mice at the MTD (i.p., 0.2 mg kg(-1)), high peak plasma concentrations of SJG 136 were seen ( C (max) = 336 nM) at 30 min after dosing. A calculated terminal t (1/2) of 0.98 h and AUC of 0.34 microM.h resulted in a clearance rate of 17.7 ml min(-1) kg(-1). The PBD dimer binds only moderately to proteins (65-75%), and in vitro cytotoxicity studies confirmed IC(50) values of 4-30 nM with a panel of human cell lines. This finding demonstrates that plasma concentrations achieved in the mouse are substantially higher than those required to elicit an anti tumour response in vitro. This report forms an important phase in the pre clinical characterization of the compound. PMID- 15122071 TI - In vitro characterization of the biotransformation of thiocoraline, a novel marine anti-cancer drug. AB - Thiocoraline is a potent new marine anti-cancer drug in vitro, which will be tested in phase I clinical studies shortly. To assess the biotransformation and the potential implications for human pharmacology and toxicology, the in vitro metabolism of thiocoraline was characterized using human plasma, human liver preparations, cytochrome P450 (CYP) and uridine diphosphoglucuronosyl transferase (UGT) supersomes and human cell lines. Thiocoraline is significantly metabolized by enzymes present in human plasma; t (1/2) shifted from 25.2 h in phosphate buffered saline to 4.3 h in human plasma. Using CYP supersomes it was shown that thiocoraline is mainly metabolized by CYP3A4, with CYP1A1, CYP2C8 and CYP2C9 playing a minor role in the biotransformation (<3%). Only minor glucuronidation was observed for thiocoraline by UGT1A1 and UGT1A9 and no glucuronidation was observed in human liver S9 fraction. In addition, no glucosidation and sulfation were observed for thiocoraline in human liver cytosol and S9 fraction. However, the metabolites formed by cytochrome P450 were further conjugated by UGT, glutathione-S-transferase (GST) and sulfotransferase (ST). In contrast to the CYP metabolism observed in supersomes, no effect could be observed from the CYP3A4 inhibitors on the cytotoxicity of thiocoraline in Hep G2 cells. However, this could be due to low CYP expression levels in the Hep G2 and IGROV-1 cell line. These results provide evidence that human CYP3A4 plays a major role in the metabolism of thiocoraline in vitro and that the metabolites formed by CYP are conjugated by the phase II enzymes UGT, ST and GST. PMID- 15122073 TI - A phase 1 study of OSI-211 given as an intravenous infusion days 1, 2, and 3 every three weeks in patients with solid cancers. AB - PURPOSE: To define the maximum tolerated dose (MTD), recommended phase II dose (RD) and dose limiting toxicity (DLT) of liposomal lurtotecan, OSI-211 (formerly known as NX211), given as a short intravenous infusion on days 1, 2, and 3 every three weeks. EXPERIMENTAL DESIGN: Thirty-seven patients were enrolled and treated in a dose escalation study from a starting dose of 0.15 mg/m(2) daily x 3 to 2.1 mg/m(2) daily x 3. Detailed pharmacokinetic analyses of blood were done on both days 1 and 3 of the first cycle and toxicity was monitored. RESULTS: Two MTDs were defined; one for patients defined as minimally pretreated and one for those heavily pretreated. Dose limiting toxicity was myelosuppression: primarily thrombocytopenia although neutropenia was also noted. The MTD was 2.1 mg/m(2)/d (total dose of 6.3 mg/m(2)) in minimally pretreated patients and 1.8 mg/m(2)/d (5.4 mg/m(2) total dose) in heavily pretreated patients. Pharmacokinetics revealed that AUC and C (max) increased with dose and were significantly higher than that of free lurtotecan (AUC approx. 100 fold higher). The half life and duration of the active lactone form were also significantly longer than historical data on free drug. Two partial responses were seen, one each in a patient with breast and ovarian cancer. CONCLUSIONS: Two Phase II recommended doses were established for OSI-211 given as a daily x 3 schedule every three weeks. The recommended phase II dose is 1.8 mg/m(2) daily x 3 for minimally pretreated patients and 1.5 mg/m(2) for those heavily pretreated. Phase II studies should be initiated in sensitive tumours. PMID- 15122072 TI - A novel mammalian cell-based approach for the discovery of anticancer drugs with reduced cytotoxicity on non-dividing cells. AB - A key asset of cytotoxic drugs in cancer therapeutics is their ability to discriminate between proliferating and mitotically inert cells and eliminate preferentially neoplastic ones. We have designed a high throughput-compatible mammalian cell-based assay for the discovery of cytotoxic drugs, which selectively kill proliferation-competent target cells. This cytotoxic drug discovery assay is based on a transgenic CHO-K1-derived cell line engineered for a conditional G1-specific growth arrest following tetracycline-responsive overexpression of the human cyclin-dependent kinase inhibitor p27(Kip1). The CHO derived cell line CHO-p27(Kip1) shows wild type proliferation rates and can be expanded in the presence of tetracycline antibiotics when p27(Kip1) expression is repressed. Upon withdrawal of regulating antibiotics CHO-p27(Kip1) differentiates into a 1:1 mixed population consisting of two different proliferation phenotypes: (i) a G1-arrested cell population induced by heterologous expression of p27(Kip1) which mimics mitotically inactive terminally differentiated cells and (ii) a proliferation-competent cell population which eliminated the p27(Kip1) expression unit and imitates neoplastic cell characteristics. Addition of chemical or metabolic libraries to CHO-p27(Kip1) populations cultivated in tetracycline-free medium followed by scoring for cell viability will reveal cytotoxic drug candidates associated with a high viability ratio of proliferation competent/arrested populations. We have validated the cell-based cytotoxic drug discovery assay using the clinically licensed cancer drugs mitomycin C, doxorubicin, etoposide and 5-fluorouracil. Comparative proof-of-concept studies showed that these top-prescribed cancer therapeutics preferentially eliminate proliferating cells while showing less interference with the viability of G1 arrested cell populations. These results demonstrate the CHO-p27(Kip1)-based cytotoxic drug finder technology is ready-to-apply for high throughput screenings of chemical as well as metabolic libraries to discover novel cancer therapeutics which show reduced cytotoxicity on terminally differentiated cells. PMID- 15122074 TI - Phase I trial of vinorelbine and diphenylhydantoin in patients with refractory carcinoma. AB - The anti-epileptic diphenylhydantoin (DPH; Dilantin) selectively enhances the in vitro cytotoxicity of vinca microtubule poisons in both parent sensitive and multi-drug resistant (MDR) human tumor cells. The in vivo clinical activity of this combination has not been fully evaluated. PURPOSE: To determine the maximum tolerated dose (MTD), dose-limiting toxicities, and preliminary antitumor activity of the combination of intravenous (IV) bolus vinorelbine (VRL) and oral diphenylhydantoin (DPH) in patients (pts) with refractory solid tumors. METHODS: Cohorts of 3-6 pts with refractory cancer were treated with escalating doses of weekly IV bolus VRL (I -20.0 mg/m(2); II -22.5 mg/m(2); III -25.0 mg/m(2); IV 27.5 mg/m(2); V -30.0 mg/m(2); VI -32.5 mg/m(2)) combined with a fixed oral dose of DPH (400 mg/day) until MTD or progression. During each 35 day cycle, pts received DPH 400 mg/day administered orally on Days -6 to Day +22 and weekly IV bolus infusion of VRL on Days +1, +8, +15, and +22. The cohort treated at the MTD was expanded to further define toxicity. RESULTS: A total of 25 evaluable pts. (9 men; 16 women) were treated with VRL and DPH at dose levels I ( n = 5), II ( n = 3), III ( n = 2), IV ( n = 3), V ( n = 7) and VI ( n = 5) in 5 week cycles over a 16 month period. Dose limiting toxicity occurred at dose level VI (VRL 32.5 mg/m(2)) and included grade 3 leukopenia ( n = 2), grade 3 neutropenia ( n = 1) and grade 4 neutropenia ( n = 1) occurring within the first cycle of treatment. There were no responses, however 9 pts had stable disease of variable duration (8 56 weeks) and received a median of 2 cycles of treatment (range 2-14). CONCLUSION: Intravenous bolus administration of VRL and oral administration of a fixed dose of DPH was well tolerated according to the schedule reported here. Although there were no responses, several patients had prolonged disease stabilization. The recommended phase II dose of VRL when used in this combination is 30 mg/m(2). PMID- 15122075 TI - Pharmacogenetics of tipifarnib (R115777) transport and metabolism in cancer patients. AB - The purpose of this study was to perform exploratory relationships between the pharmacokinetics of the farnesyl transferase inhibitor, tipifarnib (R115777, Zarnestra) and allelic variants of genes coding for ATP binding-cassette transporters and drug-metabolizing enzymes. Twenty-eight patients with advanced solid tumors were treated with tipifarnib administered orally at a dose of 200 or 300 mg. Blood samples were collected for pharmacokinetics and genotyping of 10 variants in genes encoding P-glycoprotein (ABCB1), cytochrome P450 isozymes CYP3A4 and CYP3A5, and UDP glucuronosyltransferase isozyme UGT1A1. The homozygous T -allele of ABCB1*8 (1236C > T ) was associated with a trend for a higher area under the curve of tipifarnib as compared to patients with only one or no variant alleles [mean (+/-SD), 5,303 +/- 1,620 ng.h/mL vs. 3,619 +/- 1,275 ng.h/mL; P = 0.047). No statistically significant differences were observed with any other genetic variant ( P > 0.15). Overall, this study indicates that ABCB1 genotype might be correlated with tipifarnib pharmacokinetics, although considerable overlap in exposure measures between genotype groups was observed. PMID- 15122076 TI - Phase I/II trial of gemcitabine plus docetaxel in advanced non small cell lung cancer (NSCLC). AB - INTRODUCTION: The poor prognosis of non small cell lung cancer (NSCLC), as well as the significant toxicity from many conventional cytotoxic regimens warrants the investigation of combinations of new active agents for treatment. This is a phase I-II (dose-finding, efficacy, and toxicity) study of docetaxel + gemcitabine in patients with stage IIIB-IV NSCLC without prior systemic therapy. PATIENTS were treated in cohorts of 3 with alternating increasing doses of docetaxel and gemcitabine at each level. PATIENTS: Fifty patients were entered, of which 49 were eligible including 28 males and 21 females; 15 stage IIIB and 34 stage IV; median age 57 yrs (35-74). RESULTS: The Maximum Tolerated Dose (MTD) was docetaxel 60 mg/m(2) day 1 and gemcitabine 750 mg/m(2) d1 & 8, every 21 days. The overall response rate is 20%. Eight patients are not formally assessable for response due to early discontinuation or loss to follow-up and are considered to have progressive disease. The median time to progression (TTP) is 3.5 months (1 25), with 11 pts with at least 7 months TTP. There were 10 pts (20%) with a partial response (PR); 18 (37%) maintained stable disease; 21 (43%) had progressive disease (PD) or were not assessable. TOXICITY: Forty-nine patients are evaluable for assessment for toxicity: Grade (Gr) 3/4 toxicity was documented thus: 14 with neutropenia, 1 with anemia, 1 with nausea, 2 liver function, 2 dyspnea, 2 fatigue, 1 allergy, 1 neurologic. CONCLUSION: This regimen is well tolerated and results in phase I-II testing in this patient population warrant further consideration of the study of docetaxel + gemcitabine for advanced NSCLC. PMID- 15122077 TI - Phase I dose escalation trial of feverfew with standardized doses of parthenolide in patients with cancer. AB - PURPOSE: Feverfew is a botanical product that contains parthenolide. Parthenolide has in vitro and in vivo anti-tumor and anti-angiogenic activity. Feverfew has been used extensively without any formal pharmacokinetic analysis. A Phase I trial was conducted to evaluate the pharmacokinetics and toxicity of parthenolide given as a component of "feverfew." PATIENTS AND METHODS: Feverfew (Tanacet trade mark ) was administered as a daily oral tablet in a 28-day cycle. A starting dose of 1 mg per day was explored with subsequent dose escalations to 2, 3, and 4 mg. Assessment of plasma pharmacokinetics was performed on patients accrued to the trial. Solid phase extraction and mass spectroscopy were used to evaluate parthenolide plasma concentrations. The limit of detection for parthenolide in plasma was 0.5 ng/ml. Patients were evaluated for response after every two cycles. RESULTS: Feverfew given on this schedule had no significant toxicity, and the maximum tolerated dose was not reached. When parthenolide was administered at doses up to 4 mg as a daily oral capsule in the feverfew preparation, there was not detectable concentration in the plasma. Because of this, parthenolide pharmacokinetics were not able to be completed. CONCLUSION: Feverfew, with up to 4 mg of parthenolide, given daily as an oral tablet is well tolerated without dose-limiting toxicity, but does not provide detectable plasma concentrations. Purification of parthenolide for administration of higher doses will be needed. PMID- 15122078 TI - Phase I trial of combined irinotecan and oxaliplatin given every three weeks to patients with metastatic colorectal cancer. AB - BACKGROUND: Both irinotecan and oxaliplatin are active agents in the treatment of patients with metastatic colorectal cancer (MCC). There is a strong preclinical rationale for combining these two agents. We sought to determine the dose limiting toxicity (DLT) and the maximum tolerated dose (MTD) of combined irinotecan and oxaliplatin given every three weeks. METHODS: Cohorts of patients with MCC previously treated with 5-fluorouracil received escalating doses of irinotecan (150, 175, and 200 mg/m(2)) and a fixed dose of oxaliplatin (130 mg/m(2)), both given intravenously every 3 weeks. DLT was evaluated within the first course of treatment. Objective responses were evaluated every two courses and were confirmed at least four weeks later. RESULTS: Fourteen patients were treated and evaluated for toxicity. The DLT was neutropenia, with or without fever, and delayed recovery of neutrophil counts was frequent (13 courses in six patients). Other toxic effects (peripheral neuropathy, nausea, vomiting, diarrhea, and fatigue) were mild to moderate. Among 13 patients evaluable for activity, four achieved partial responses and nine had stable disease. CONCLUSION: The combination of irinotecan and oxaliplatin is safe and apparently active in the treatment of MCC patients. The recommended dose for phase II studies is 175 mg/m(2) irinotecan plus 130 mg/m(2) oxaliplatin, given every 3 weeks. Neutropenia and delayed recovery of neutrophil counts are the predominant early toxicities with this schedule. PMID- 15122079 TI - Phase II trial of flavopiridol, a cyclin dependent kinase inhibitor, in untreated metastatic malignant melanoma. AB - PURPOSE: To test the activity of the cyclin dependent kinase (cdk) inhibitor flavopiridol in malignant melanoma, a disease with frequent abnormalities of the cyclin dependent kinase system. PATIENTS AND METHODS: Patients had histologically proven, unidimensionally measurable malignant melanoma, incurable by standard therapy. Prior adjuvant immunotherapy was allowed, but patients were otherwise untreated for advanced disease. Flavopiridol was administered at a dose of 50 mg/m(2) IV over 1 hour daily x 3 days every 3 weeks. Patients were assessed for response every 2 cycles. RESULTS: 17 patients were accrued over 5 months. No objective responses were documented in the 16 patients evaluable for response. Seven patients (44%) had stable disease after 2 cycles, with a median of 2.8 months (range 1.8-9.2). The most common treatment-related non-hematologic toxicities were diarrhea (82%), nausea (47%), fatigue (41%), anorexia (35%) and vomiting (29%). Most treatment-related toxicities were mild, except for diarrhea (grade 3 in 3 patients, grade 4 in 1 patient), nausea (grade 3 in 1 patient) and tumor pain (grade 3 in 1 patient). Hematologic toxicities were minimal, none worse than grade 2. Eighty-eight percent of patients received >/=90% planned dose intensity; 2 patients had dose reductions for gastrointestinal (GI) toxicity. CONCLUSIONS: Flavopiridol is well tolerated at the dose regimen used in this study, with an acceptable (primarily GI) toxicity profile. Although 7 of the 16 patients had stable disease ranging from 1.8 to 9.2 months in duration, there was no evidence of significant clinical activity in malignant melanoma by objective response criteria. PMID- 15122080 TI - 9-Aminocamptothecin (9-AC) given as a 120-hour continuous infusion in patients with advanced adenocarcinomas of the stomach and gastroesophageal junction: A phase II trial of the University of Chicago phase II consortium. AB - 9-Aminocamptothecin (9-AC) is a water-insoluble camptothecin derivative that demonstrated broad activity in pre-clinical studies. In vitro, greater anti-tumor efficacy can be achieved with prolonged administration. A minor response was observed in gastric cancer in a phase I study. We conducted a phase II study of 9 AC in 15 patients with previously untreated metastatic gastric cancer and adenocarcinoma of the gastroesophageal junction. 9-AC was administered at a dose of 25 microg/m(2)/h over 120 hours (3000 microg/m(2) over 5 days) on two consecutive weeks every 21 days. Fourteen patients were evaluable for response. There were no objective responses. Three patients had stable disease lasting a median of 3.4 months (range 1.6-4.3 months). Median time to progression was 1.4 months; median survival was 5.2 months. Grade 3 neutropenia developed in 20% of patients, and anemia in 7%. Grade 3 nausea and fatigue each developed in 7% of patients. We conclude that 9-AC given by 120-hour continuous infusion demonstrates no clinical activity in patients with metastatic gastric cancer. PMID- 15122081 TI - Phase II study on 9-nitrocamptothecin (RFS 2000) in patients with advanced or metastatic urothelial tract tumors. AB - OBJECTIVE: To investigate the antitumor activity and the safety of RFS2000, an oral topoisomerase I inhibitor, in patients with advanced or metastatic urothelial tract tumors refractory to one prior chemotherapy regimen. PATIENTS AND METHODS: Eligible patients were to have failed first line treatment for advanced or metastatic disease. Patients received RFS2000 as one daily oral intake at the dose of 1.5 mg/m(2)/day according to a "5 days on/2 days" off schedule continuously. One cycle was arbitrarily defined as a 3 week period. Sufficient oral fluid intake to prevent cystitis previously described in phase I trials with RFS2000 was recommended. Gehan design was used for sample size determination. Anti-tumor activity was evaluated according to the RECIST criteria and toxicity according to CTC version 2. RESULTS: Twenty patients received a total of 57 cycles (range 1-8). Grade 3-4 toxicity was observed in 10 patients requiring dose or schedule modifications. Hematological grade 3-4 toxicity was observed in 16% of the cycles. Only one patient experienced a partial response. CONCLUSIONS: RFS2000 could be administered orally as a "5 days on/2 days off" schedule continuously with a median dose intensity of 90.6% with an acceptable toxicity profile. However, RFS2000 did not exert significant activity in patients with advanced/metastatic urothelial tract tumors failing prior chemotherapy. The results of this study do not suggest further investigation of RFS2000 at the present dose and schedule for the treatment of urothelial tract tumors in this refractory population. PMID- 15122082 TI - A phase II study of high-dose 24 hour continuous infusion 5-FU and leucovorin and low-dose PALA for patients with advanced pancreatic adenocarcinoma: a Southwest Oncology Group Study. AB - PURPOSE: The purpose of this phase II multi-institutional study was to define the efficacy and toxicity of infusional 5-FU in combination with PALA and leucovorin in patients with advanced pancreatic cancer. PATIENTS AND METHODS: Patients were required to have histologically confirmed pancreatic cancer that was locally advanced, unresectable or disseminated. The treatment regimen consisted of weekly 5-FU 2600 mg/m(2) given concurrently with leucovorin at 500 mg/m(2). Both drugs were administered by 24-hour continuous infusion. PALA was administered 24 hours prior to the administration of 5-FU/LV at a dose of 250 mg/m(2) IV over 15 minutes weekly. Patients were continued on the assigned treatment regimen until progression of disease, unacceptable toxicity, or the patient declined further therapy. RESULTS: This study accrued 30 patients. Four of these patients were ineligible. All 26 eligible patients were evaluated for toxicity. One patient had inadequate assessment of response and was considered a non-responder. Three of the twenty-six eligible patients had partial responses, for a response rate of 12% (95% confidence interval 2% to 30%). All 26 eligible patients have died and the median overall survival was 7 months (95% confidence interval: 5.2 to 9 months). Four patients experienced grade 4 toxicities, including bilirubin increase (2 patients), vomiting (1 patient) and non-local skin ulceration (1). Two patients discontinued therapy due to toxicity. CONCLUSION: The dual modulation of 5-FU with PALA and leucovorin in the dose and schedule used here, has a response rate similar to other single agents in pancreatic cancer and can result in some long term survival while having relatively mild toxicity. PMID- 15122085 TI - Cellular mechanisms of insulin secretion. PMID- 15122086 TI - Mechanisms of glucose sensing and multiplicity of glucose sensors. PMID- 15122083 TI - Evaluation of irofulven (MGI-114) in the treatment of recurrent or persistent endometrial carcinoma: A phase II study of the Gynecologic Oncology Group. AB - OBJECTIVE: This multi-center phase II trial was conducted by the Gynecologic Oncology Group to evaluate the activity and toxicity of irofulven in patients with previously treated adenocarcinoma of the endometrium. METHODS: Eligible patients had documented recurrent or persistent endometrial carcinoma after receiving definitive locoregional therapy, and were required to have measurable disease, performance status of 0-2, and adequate bone marrow, hepatic and renal functions prior to study entry. Patients were allowed one prior chemotherapy regimen. The initial dose of irofulven was 11 mg/m(2)/day for four days administered intravenously. Cycles were repeated every 28 days. Doses were escalated or reduced based on previous cycle toxicity. RESULTS: Twenty-five patients were enrolled onto the trial. There was one (4%) confirmed complete response. Seven (28%) patients had stable disease, with a median duration of 10.4 (range: 4.4-21.6) months. Patients received a median of one (range: 1-5) cycle of protocol treatment. There were three early treatment-related deaths due to renal failure and severe electrolyte disturbances. Two patients experienced grade 4 hematologic adverse effects. CONCLUSION: Irofulven administered at the dose and schedule used in this trial was minimally active and resulted in significant toxicity. PMID- 15122087 TI - Gastro-intestinal hormones GIP and GLP-1. PMID- 15122088 TI - [Pathophysiology of glucagon secretion]. PMID- 15122089 TI - Pathophysiology of insulin secretion. AB - Defects in pancreatic islet beta-cell function play a major role in the development of diabetes mellitus. Type 1 diabetes is caused by a more or less rapid destruction of pancreatic beta cells, and the autoimmune process begins years before the beta-cell destruction becomes complete, thereby providing a window of opportunity for intervention. During the preclinical period and early after diagnosis, much of the insulin deficiency may be the result of functional inhibition of insulin secretion that may be at least partially and transiently reversible. Type 2 diabetes is characterized by a progressive loss of beta-cell function throughout the course of the disease. The pattern of loss is an initial (probably of genetic origin) defect in acute or first-phase insulin secretion, followed by a decreasing maximal capacity of insulin secretion. Last, a defective steady-state and basal insulin secretion develops, leading to almost complete beta-cell failure requiring insulin treatment. Because of the reciprocal relation between insulin secretion and insulin sensitivity, valid representation of beta cell function requires interpretation of insulin responses in the context of the prevailing degree of insulin sensitivity. This appropriate approach highlights defects in insulin secretion at the various stages of the natural history of type 2 diabetes and already present in individuals at risk to develop the disease. To date none of the available therapies can stop the progressive beta-cell defect and the progression of the metabolic disorder. The better understanding of the pathophysiology of the disease should lead to the development of new strategies to preserve beta-cell function in both type 1 and type 2 diabetes mellitus. PMID- 15122090 TI - Gluco-lipotoxicity of the pancreatic beta cell. PMID- 15122091 TI - Alteration in insulin action: role of IRS-1 serine phosphorylation in the retroregulation of insulin signalling. AB - Insulin resistance, when combined with impaired insulin secretion, contributes to the development of type 2 diabetes. Insulin resistance is characterised by a decrease in insulin effect on glucose transport in muscle and adipose tIssue. Tyrosine phosphorylation of insulin receptor substrate 1 (IRS-1) and its binding to phosphatidylinositol 3-kinase (PI 3-kinase) are critical events in the insulin signalling cascade leading to insulin-stimulated glucose transport. Modification of IRS-1 by serine phosphorylation could be one of the mechanisms leading to a decrease in IRS-1 tyrosine phosphorylation, PI 3-kinase activity and glucose transport. Recent findings demonstrate that "diabetogenic" factors such as FFA, TNFalpha, hyperinsulinemia and cellular stress, increase the serine phosphorylation of IRS-1 and identified Ser307/612/632 as phosphorylated sites. Moreover, several kinases able to phosphorylate these serine residues have been identified. These exciting results suggest that serine phosphorylation of IRS-1 is a possible hallmark of insulin resistance in biologically insulin responsive cells or tIssues. Identifying the pathways by which "diabetogenic" factors activate IRS-1 kinases and defining the precise role of serine phosphorylation events in IRS-1 regulation represent important goals. Such studies may enable rational drug design to selectively inhibit the activity of the relevant enzymes and generate a novel class of therapeutic agents for type 2 diabetes. PMID- 15122092 TI - Mouse models of insulin resistance and type 2 diabetes. PMID- 15122093 TI - [The adipocyte and insulin resistance]. PMID- 15122094 TI - [Genes regulated in muscle and adipocyte]. PMID- 15122095 TI - [Thiazolidinediones and insulin sensitivity]. PMID- 15122096 TI - The CREB family: key regulators of hepatic metabolism. PMID- 15122097 TI - [Glucagon and glucose metabolism]. PMID- 15122098 TI - Growth hormone and glucose metabolism: the model of the GH-receptor antagonists. AB - Pegvisomant is a GH analogue that includes a single amino acid substitution at position 120 that generates the GHR antagonist. Additional changes include amino acid substitutions within binding site 1 and a further modification by the addition of polyethylene glycol moieties that increase the half-life and reduce the immunogenicity of the molecule. In acromegalics, pegvisomant is the most effective treatment for normalizing the IGF-I, and pegvisomant significantly improves insulin sensitivity in patients suffering from acromegaly. However, there are simply no data available that might support a role for pegvisomant treatment in disorders in which glucose metabolism is disturbed and in which reducing GH action would be theoretically beneficial. PMID- 15122100 TI - Counterregulation to hypoglycaemia: physiology. PMID- 15122101 TI - Hyperinsulinemic hypoglycemia in adults. AB - The diagnosis of a hypoglycemic disorder requires a high level of suspicion, careful assessment of the patient for the presence of mediating drugs or a predisposing illness, and, where indicated, methodical evaluation on the basis of well-defined diagnostic criteria. The diagnostic burden is heaviest for healthy appearing persons with episodes of confirmed neuroglycopenia. Our criteria for insulin mediation of hypoglycemia are: plasma insulin >or=18 pmol (ICMA [immunochemiluminometric assay]), C-peptide >or=200pmol/L (ICMA), proinsulin >or=5pmol/L (ICMA), betaOH butyrate,40 Gy) were evaluated through the whole saliva test (WST) before, during and after ten weeks of treatment with pilocarpine hydrochloride (5 mg t.i.d.). Hyposalivationrelated symptomatology was assessed before and at every week by means of a questionnaire with an ordinal scale ranging from 0-10. Salivary production values recorded at the end of the study and those obtained before treatment were compared by means of student's t test. A paired Wilcoxon test was used to compare the differences in the oral symptoms, such as oral dryness, soreness, ability for speaking and swallowing before and after treatment. RESULTS: Initial WST had a mean salivary production of 0.8 cm (s.d. 0.7), with a range from 0 to 2.9 cm). After ten weeks of treatment salivary production increased to a mean of 2.24 cm (s.d. 0.7), with a range from 1.2 to 4.0 cm. There was a salivary flow increase of 64.5% (p<0.001). In addition, there were significant improvements in oral dryness, mouth comfort, ability to speak and ability to swallow (p<0.01). Adverse effects were usually minimal and they did not cause withdraw from the study in any case. CONCLUSIONS: Administration of 5-mg pilocarpine hydrochloride tablets three times daily significantly improved salivary production and clinical symptomatology secondary to radiation-induced hyposalivation with minimal side-effects. In addition, this study showed that WST is a fast, technically simple and highly reliable method to study salivary production in prospective studies. PMID- 15122123 TI - Tuberous sclerosis and its oral manifestations. A clinical case. AB - Tuberous sclerosis (TS) is a congenital anomaly in the development of the embryo which is transmitted through the autosomal dominant gene. It has various forms of clinical expression. It is classified as one of what are known as Phacomatoses (phakos stain and oma tumour), which are development anomalies that can originate tumours and/or hamartomas in the nervous system. Lesions in the nervous system are nearly always accompanied by cutaneous anomalies. In this study we introduce the case of a woman patient aged 55, diagnosed several years earlier with Tuberous Sclerosis, who attended for intraoral lesions which were clinically compatible with fibromata. These lesions of fibrous appearance occurred above all on the lower lip and in both cheeks' mucous membranes, little mentioned in the literature as a site for such a manifestation of TS. The pathological anatomy suggested lesions compatible with Angiomyolipoma. In addition, the data referring to TS are reviewed; its implications for the mouth are described; and histopathological results are used to examine the significance of the word Angiomyolipoma. PMID- 15122122 TI - Spectrum of oral manifestations of HIV/AIDS in the Perm region (Russia) and identification of self-induced ulceronecrotic lingual lesions. AB - OBJECTIVE: To study the frequency and spectrum of oral manifestations of HIV infected drug-users in the Perm region. SUBJECTS: 104 seropositive HIV-infected drug-users (69 male, 35 female; ages 15 to 32 years; 13 co-infected with hepatitis viruses) and 13 AIDS-infected drug-users (7 male, 6 female; ages 16 to 37 years; 12 co-infected with hepatitis viruses). RESULTS: The most frequent forms of oral mucosal lesions in the HIV-infected group -- candidiasis (32.7%), herpetic lesions (15.4%), cheilitis glandularis (3.9%), recurrent aphthous stomatitis (2%). Regional lymphadenopathy was observed in 31% cases. The ulceronecrotic oral mucosal lesions were seen in the sublingual region and tongue in 11.5% patients and manifested with pain, dysarthria, dysphagia, and dysgeusia. These lesions were found in drug-users who injected the opioids sublingually. AIDS patients had oral candidiasis (84.6%), herpetic lesions (53.8%), recurrent aphthous stomatitis (15.4%) and cheilitis glandularis (7%). All AIDS-patients had severe xerostomia, and 15.4% had unilateral or bilateral swelling of the parotid glands. Generalized ulceronecrotic gingivostomatitis was found in 50% of the patients but the sublingual ulceronecrotic lesions were not identified. CONCLUSIONS: 1. The spectrum of oral cavity lesions of HIV/AIDS patients in Perm region is widespread enough. 2. Dissemination of oral cavity lesions is increasing in proportion of disease progression. 3. Dental care of HIV/AIDS patients should include periodic oral examinations to monitor their disease progression and to alleviate symptoms of oral opportunic and neoplastic diseases, to improve the life-style of the patients infected with HIV. PMID- 15122124 TI - Considerations and differences in the treatment of a fused tooth. AB - Dental malformations relating to bigeminism, fused teeth, etc., have a prevalence of 0.5-5%, depending on geographic, racial or genetic factors. This in turn may influence the treatment to be provided in each case. Anomalies in the shape, size and number of teeth are more common in the so-called pediatric genetic syndromes, many of which are associated to mental retardation. The present study describes two clinical cases of similar dental fusion involving a central incisor and comprising different and individualized treatment plans due to the important differences in the characteristics of each patient. The first case corresponded to a healthy 9-year-old boy with a fused upper central incisor and a supernumerary incisor in the second quadrant. The fused tooth presented two independent and converging roots with a single pulp chamber. Endodontic treatment was provided, with dental sectioning and composite reconstruction of the crown. Orthodontic management was reserved for a second stage. The second patient was a 27-year-old male with severe mental retardation of uncertain origin, gingival hyperplasia, a large bilobular upper central incisor, and a total lack of oral hygiene. Treatment in this case was limited to surgical resolution of the periodontal problem and the introduction of an exhaustive prevention program involving instructions for oral hygiene (with implication of the parents and care takers). In conclusion, the development of a management plan in these two cases of fused teeth required due consideration of the degree of patient cooperation, which was severely limited in one case -- thereby causing treatment to differ greatly from one individual to another. PMID- 15122126 TI - Immediate implants after extraction. A review of the current situation. AB - Immediate implants are positioned in the course of surgical extraction of the tooth to be replaced. The percentage success of such procedures varies among authors from 92.7-98.0%. The main indication of immediate implantation is the replacement of teeth with pathologies not amenable to treatment. Its advantages with respect to delayed implantation include reduced post-extraction alveolar bone resorption, a shortening of the rehabilitation treatment time, and the avoidance of a second surgical intervention. The inconveniences in turn comprise a general requirement for membrane-guided bone regeneration techniques, with the associated risk of exposure and infection, and the need for mucogingival grafts to seal the socket space and/or cover the membranes. The surgical requirements for immediate implantation include extraction with the least trauma possible, preservation of the extraction socket walls and thorough alveolar curettage to eliminate all pathological material. Primary stability is an essential requirement, and is achieved with an implant exceeding the alveolar apex by 3-5 mm, or by placing an implant of greater diameter than the remnant alveolus. Esthetic emergence in the anterior zone is achieved by 1-3 mm sub-crest implantation. Regarding guided regeneration of the alveolar bone, the literature lacks consensus on the use of membranes and the type of filler material required. While primary wound closure is desirable, some authors do not consider it to be of great relevance. PMID- 15122125 TI - Transformation of proliferative verrucous leukoplakia to oral carcinoma: a ten years follow-up. AB - The authors present a case of proliferative verrucous leukoplakia (PVL) in a 78 year-old man. It was initially presented as leukoplakia on the tongue but a microscopic investigation in 1991 revealed it to be a mild epithelial dysplasia. After 5 years of follow-up, the lesion presented changes in size and location, and a recidivant behavior. In 1996, a red granular and indurated area that appeared on the tongue was found to be a microinvasive squamous cell carcinoma when microscopically investigated. After a review of the clinicopathologic behavior of this entity, the authors concluded that it was a typical PVL, whose diagnosis is difficult and retrospective, as indicated by others. The authors emphasize the importance of periodic detailed clinical and histological examination of this type of lesions in order to detect early signs of malignancy. PMID- 15122127 TI - Development of tooth germ heterotopically grafted within the ear skin. An histological study in the rat. AB - The main goal of this study was the analysis of the developmental potentiality of tooth germ from late bell stage on, after its heterotopic placement within the skin. Teeth germs of newborn rats were grafted within a skin pouch of the ear of adult rats. Seven to fourteen days after grafting, dental germs developed normal dental structures in which ameloblasts and odontoblasts were well differentiated. Twenty to forty-one days after graft, the inflammatory host reaction destroyed the dental developed tissues by cell infiltration. The dentin of the grafts was of osteoid characteristics, and its size increased depending on grafting time until the complete substitution of all dental tissues. This atypical dentin showed several degrees of polymerisation from collagen fibres smooth dentin devoid near the graft a to fibres rich dentin far from the dental germ. Present results suggest that this type of dental graft could be a valuable model to study the self-development of dental tissues and the reactive mechanisms taking place after dental injuries. PMID- 15122128 TI - Complications of ambulatory oral surgery in patients over 65 years of age. AB - Serious systemic disorders such as hypertension, cerebrovascular or heart disease, diabetes and psychiatric problems are common in elderly patients, and lead to the prescription of different drugs. This may in turn influence oral health, and the dentist should be familiarized with these situations when providing dental treatment in elderly patients. A retrospective study was made of 196 patients over age 65 years to evaluate the type of ambulatory surgery performed under locoregional anesthesia, taking into account the presence of background systemic pathology, multiple drug therapy, oral and dental health, the cause of consultation and the type of anesthesia used, relating these parameters to the development of intra- or postoperative systemic and/or local complications. Some systemic disease was documented in 88% of the patents-- hypertension being the most frequent disorder (in 45% of subjects with systemic disease). On the other hand, 78% of the patients used some medication, and 77% presented for hard-tissue treatment (tooth extractions, bone remodeling, etc.); 61% of all treatments comprised the removal of root fragments (54% of all hard tissue interventions), symptomatic third molars (15%) or other dental inclusions. Only mild or moderate complications were recorded (13%) -- either local (n = 25) or systemic (n = 1). No significant relation was observed between the development of intra- or postoperative complications and the type of treatment provided or the medication used by these patients. Only diabetes was associated with a significant increase in intra- and postoperative local complications (p<0.003). PMID- 15122129 TI - Plexiform neurofibroma of the cheek mucosa. A case report. AB - The case reported deals with a solitary plexiform neurofibroma affecting the cheek submucosa. Neurofibroma is an uncommon tumor which rarely appears in oral cavity but it represents the most common neurogenic tumor. Furthermore, plexiform variety is less frequent. Clinically, oral neurofibromas usually appears as anodyne and asintomatic lesions. Sometimes, they produce nervous compression. In this case, tumor is big but asintomatic. There is no definitive radiologic image. It has association with polyglandular syndromes and phacomatosis. The treatment of choice is excision. There are doubts of the surgical results so that some authors are looking for new non-surgical treatments. The clinical characteristics, epidemiology, diagnosis and treatment are described as soon as a bibliographic revision. PMID- 15122130 TI - Cartilage in the walls of odontogenic keratocyst. AB - There are seven published cases in world literature on cartilage in the walls of odontogenic keratocysts. Herein is presented one further case with keratin inclusions in the cystic wall, which also bears a cartilaginous component. X rays, clinical images and pathohistological images are included. PMID- 15122131 TI - Stafne's cavity. PMID- 15122132 TI - Osteosarcoma of the jaws. PMID- 15122133 TI - Patient-controlled ropivacaine analgesia after arthroscopic subacromial decompression. AB - PURPOSE: To evaluate the efficacy of a subacromial patient-controlled analgesia (PCA) infusion of 0.2% ropivacaine versus saline for postoperative pain control following arthroscopic shoulder surgery. TYPE OF STUDY: Double-blind prospective randomized study. METHODS: A prospective, randomized double-blind study was performed on a consecutive group of 24 patients. All patients had arthroscopic subacromial decompression. Ropivacaine was chosen as the study drug due to the association of cardiac toxicity with the use of bupivacaine. The surgeon, anesthesiologist, and the operating room staff were blinded to the randomization. Immediately before surgery, the pharmacy staff randomized the type of drug infusion. Patients were asked to record their pain score using a visual analog scale (VAS) along with the amount of hydrocodone consumption for the first 2 days after surgery. RESULTS: Nineteen patients completed the study. Ten patients received a subacromial infusion of 0.2% ropivacaine at 5 mL per hour with a bolus dose of 2 mL at a 15-minute lockout period via a microjet PCA pump (group I). Nine patients received saline in the same experimental conditions (group II). The use of a PCA ropivacaine infusion (group I) resulted in a significant reduction of postoperative pain by 34% as measured by VAS scale (P <.05), but no changes in the amount of hydrocodone consumption. CONCLUSIONS: The use of subacromial 0.2% ropivacaine PCA infusion provided effective postoperative pain control. LEVEL OF EVIDENCE: Level I. PMID- 15122134 TI - Arthroscopic versus open treatment of Bankart lesion of the shoulder: a prospective randomized study. AB - PURPOSE: The purpose of this study was to compare the results of arthroscopic and open repair of isolated Bankart lesions of the shoulder using metallic suture anchors. TYPE OF STUDY: Prospective randomized clinical study. METHODS: Sixty patients with traumatic anterior shoulder instability underwent a surgical repair of an isolated Bankart lesion. The patients were divided into 2 groups of 30 patients each. In group 1, an arthroscopic repair was performed, and in group 2, an open procedure was performed. The groups were homogeneous for gender, age, dominance, number of dislocations, time elapsed between first dislocation and surgery, and pathologic findings. In all cases of both groups, the lesion was repaired using metallic suture anchors carrying nonabsorbable braided sutures. Postoperative rehabilitation was the same for the 2 groups. Two years' follow-up evaluation included Constant and Rowe shoulder scores. Statistical analysis of data was performed using an unpaired t test (significance for P <.05). RESULTS: No recurrence of dislocation of the involved shoulder has been reported in either group. Follow-up Constant and Rowe scores of the 2 groups were not significantly different. The only significant difference seen between the 2 groups was for range of motion evaluation with the Constant score. The mean value for group 1 (39.6 +/- 0.8) was significantly greater (P =.017) than that for group 2 (37.8 +/ 2.0). CONCLUSIONS: Arthroscopic repair with suture anchors is an effective surgical technique for the treatment of an isolated Bankart lesion. Open repair does not offer a significantly better 2-year result in terms of stability, and furthermore, can negatively affect the recovery of full range of motion of the shoulder. LEVEL OF EVIDENCE: Level I. PMID- 15122135 TI - Lateral retinacular release: a survey of the International Patellofemoral Study Group. AB - PURPOSE: The purpose of this investigation was to determine current views regarding lateral release among experienced knee surgeons with a specific interest in the patellofemoral joint. TYPE OF STUDY: Scientific survey. METHODS: A questionnaire was developed and mailed to all members of an international group with a specific interest in disorders of the patellofemoral joint. Frequencies and percentages of responses were calculated for each question to determine surgeon consensus. We measured agreement among responses using the kappa statistic. This provided an indication of consistency for each question as well as correlation among the responses to different questions. RESULTS: The survey response rate was 60%. Isolated lateral release was estimated to account for only 1 to 5 surgical cases per respondent per year, or 2% of cases performed annually. In the setting of arthroscopy or exploration, 74% of respondents believed that lateral release calls for specific informed consent. Strong consensus was found that objective evidence is needed to justify lateral release, but agreement was poor as to what clinical evidence provides the most appropriate indication for the procedure. CONCLUSIONS: Even among experienced knee surgeons with a special interest in diseases of the patellofemoral articulation, isolated lateral release is rarely performed. Strong consensus was found that isolated lateral release should not be undertaken without prior planning in the form of objective clinical indications and preoperative informed consent. LEVEL OF EVIDENCE: Level V. PMID- 15122137 TI - Arthroscopic evaluation of the articular cartilage after anterior cruciate ligament reconstruction: a short-term prospective study of 105 patients. AB - PURPOSE: Several reports have shown the progression of degenerative osteoarthritis after anterior cruciate ligament (ACL) reconstruction. No report has been published about early cartilage change after ACL reconstruction. The purpose of this study was to evaluate the articular cartilage after ACL reconstruction in a short postoperative period by arthroscopy. TYPE OF STUDY: Case series. METHODS: We examined the status of articular cartilage of 105 patients who received ACL reconstruction and second-look arthroscopy. Cartilage lesion was evaluated arthroscopically in the 6 articular surfaces independently, and these features were classified by modified Outerbridge's classification. We compared the articular cartilage at reconstruction and at second-look arthroscopy. RESULTS: A significant worsening of the status of the articular cartilage was seen after ACL reconstruction. This worsening was seen at all articular surfaces except the lateral femoral condyle. Most of the change involved softening or fibrillation. Anterior laxity and meniscal lesion had no correlation with a progression of degenerative change of articular cartilage. Patient's age influenced the progression of articular cartilage damage after reconstruction significantly in our cases. CONCLUSIONS: The status of articular cartilage was significantly worsened after ACL reconstruction. Potent risk factors causing articular cartilage damage include female gender and age of 30 years or older. LEVEL OF EVIDENCE: Level IV. PMID- 15122136 TI - Screening for arthrofibrosis after anterior cruciate ligament reconstruction: analysis of association with human leukocyte antigen. AB - PURPOSE: Arthrofibrosis represents a severe complication of trauma and reconstructive joint surgery because of generalized connective tissue proliferation resulting in painful joint stiffness. It often appears stereotypical in terms of its clinical and pathologic features, comprising excess deposition of extracellular matrix proteins such as collagen type I, III, and VI and proliferation of fibroblasts. However, trauma and surgery around joints does not always lead to fibrosis, suggesting a genetic predisposition. For a number of autoimmune diseases, strong associations have been described. The objective of the study was to investigate whether an association of HLA (human leukocyte antigen) with primary arthrofibrosis exists. TYPE OF STUDY: Retrospective cohort study. METHODS: Seventeen patients with primary arthrofibrosis after autologous anterior cruciate ligament (ACL) reconstruction were identified and clinically reviewed. Blood samples were taken, and DNA was isolated by column extraction method. DNA samples were typed for the loci HLA-A, -B, -C, -DRB1, and -DQB1. Results were compared with the frequencies of allelic groups as determined for the caucasoid population. RESULTS: HLA-Cw*07 was significantly less often found in the patient group than in the general population (P =.022). The opposite effect was seen for Cw*08, which was found in 17.6% of the patient group but only in 3.8% of the reference group (P =.045). A significant difference was also seen for DQB1*06, because 23.5% of the patients but 48.6% of the reference group possessed an allelic variant of this group (P =.048). However, according to the relatively small number of patients, a statistical bias cannot be excluded. CONCLUSIONS: A possible link may exist between arthrofibrosis and HLA-Cw*07- and DQB1*06-negative as well as Cw*08-positive individuals. Further investigation is necesessary to confirm or vitiate the possible association. LEVEL OF EVIDENCE: Level IV. PMID- 15122138 TI - Expression changes of gelatinases in human osteoarthritic knees and arthroscopic debridement. AB - PURPOSE: The purpose of this study was to quantify the expression changes of gelatinase-A and -B (matrix metalloproteinase [MMP]-2 and MMP-9) in a series of chondral, meniscal, and synovial cultures of knee osteoarthritis (OA) for investigation of the possible roles of the cartilage, menisci, and synovia and the efficacy of arthroscopic debridement. TYPE OF STUDY: A biochemical study. METHODS: In 43 consecutive patients with knee OA undergoing arthroscopic debridement, we examined the amount of MMP-2 and MMP-9 in a series of chondral, meniscal, and synovial cultures. We also compared the gene expressions of MMP-2 and MMP-9 and membrane-type 1 MMP (MT1-MMP) in the chondral, meniscal, and synovial cultures using reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Latent and activated forms of MMP-2 were produced in all series of chondral, meniscal, and synovial cultures, and the levels in lesional cultures were significantly higher than those in paralesional ones (P <.001). Moreover, the latent form of MMP-9 (proMMP-9) appeared in 29 of 37 series of synovial cultures and in 13 of 40 series of meniscal cultures. In meniscal cultures after 24 hours of incubation and synovial cultures after 3 and 24 hours of incubation, the level of proMMP-9 in lesional cultures was significantly higher than that in paralesional ones (P <.001). The activated form of MMP-9 appeared in 10 of 37 series of synovial cultures, and the level in lesional cultures was significantly higher than that in paralesional ones (P <.05). Furthermore, MMP-2, -9, and MT1 MMP mRNA levels of lesional areas also showed the increased expression in RT-PCR. CONCLUSIONS: Our data confirm that tissue repair of OA is ascribable to enzymic digestion of the extracellular matrix ex vivo. When technically appropriate, arthroscopic debridement for the pathologic lesions of OA, such as meniscal tears, chondral lesions, and hypertrophic villi, may be beneficial to the process of early cases. Still, it should be carefully studied for its overall effect and mechanism in vivo. CLINICAL RELEVANCE: The effectiveness of arthroscopic debridement for the treatment of knee OA is controversial. The present study provides the possible mechanism of the positive effects of arthroscopic debridement in basic science. PMID- 15122139 TI - Arthroscopic knots: determining the optimal balance of loop security and knot security. AB - PURPOSE: The purpose of this study was to determine the optimal knot configuration that maximized both knot and loop security when tied with 2 different types of nonabsorbable, braided suture. TYPE OF STUDY: In vitro biomechanical study. METHODS: Six commonly used arthroscopic sliding knots (Duncan loop, Nicky's knot, Tennessee slider, Roeder knot, SMC knot, Weston knot) with and without a series of 3 reversing half-hitches on alternating posts (RHAPs) as well as a static surgeon's knot were tied. Two different nonabsorbable, braided sutures were used, and a total of 7 knots were tied for each possible combination of knots and sutures, for a total of 182 knots. Each knot was tied around a 30-mm circumference post to assure a consistent loop circumference of 30 mm before "locking" the complex sliding knots by tensioning the wrapping limb of the suture. Each loop was mounted on a Material Testing System machine, and its circumference was measured at a 5-N preload to assess each knot's ability to maintain a tight suture loop without slippage (loop security). Knot security was measured as the maximum force to failure at 3 mm of crosshead displacement or suture breakage during single-pull load testing. RESULTS: The surgeon's knot provided the highest force to failure and the tightest loop circumference whether tied with No. 2 Ethibond (Ethicon, Somerville, NJ) or No. 2 Fiberwire (Arthrex, Naples, FL) suture. Among the sliding knots, the Roeder knot with 3 RHAPs showed the best balance of loop security and knot security when tied with No. 2 Ethibond or No. 2 Fiberwire. Sliding knots tied without RHAPs showed low force to failure and loose suture loops whether tied with Ethibond or Fiberwire. The addition of 3 RHAPs improved knot security and, in most cases, loop security of all the sliding knots. When tying a static surgeon's knot or a sliding knot with RHAPs, using No. 2 Fiberwire increased the force to failure over comparable knots tied with No. 2 Ethibond. All knots failed by a combination of knot slippage and suture stretch. When using No. 2 Ethibond, securing most sliding knots with 3 RHAPs or tying a surgeon's knot changed the failure mechanism from knot slippage to suture stretch, suggesting that the maximum knot holding capacity of No. 2 Ethibond had been achieved when tying these knot configurations. However, even at failure forces twice that achieved with No. 2 Ethibond, suture slippage continued to occur with sliding knots with 3 RHAPs using No. 2 Fiberwire. This indicates that the maximum knot-holding capacity of No. 2 Fiberwire had not been achieved, and that further knot configurations should be tested. CONCLUSIONS: (1) A static surgeon's knot provides the best balance of loop security and knot security within the knot configurations tested in this study. (2) A sliding knot without RHAPs has both poor loop security and knot security and should not be tied. (3) The addition of 3 RHAPs improves knot security of all sliding knots tested and improves loop security of most of the sliding knots tested. (4) The addition of 3 RHAPs improves the knot security of all sliding knots to adequately resist predicted in vivo loads. (5) The Roeder knot with 3 RHAPs provides the best balance of loop security and knot security within the sliding knot configurations tested in this study regardless of suture type. (6) Tying a surgeon's knot or a sliding knot with 3 RHAPS using No. 2 Fiberwire increases knot security over the same knot tied with No. 2 Ethibond. CLINICAL RELEVANCE: This study identifies the static and sliding configurations of commonly used arthroscopic knots in order to aid the surgeon in choosing the most biomechanically effective knot for use in arthroscopic surgery. PMID- 15122140 TI - Chondrocyte viability and metabolic activity after treatment of bovine articular cartilage with bipolar radiofrequency: an in vitro study. AB - PURPOSE: Some controversy exists regarding the effects of radiofrequency (RF) probes on articular cartilage. To further elucidate these effects, we examined the chondrocyte viability and metabolic activity after treatment of fresh bovine articular cartilage with bipolar RF probes. TYPE OF STUDY: In vitro assessment. METHODS: Three fresh bovine knees served as a baseline control for chondrocyte viability, yielding 6 samples (1 from each medial femoral condyle and 1 from each lateral femoral condyle). After the baseline expected chondrocyte viability was determined, 3 additional bovine knees served as the experimental specimens for the study. Under sterile conditions, 2 different bipolar RF probes were used to treat the articular surface in a light contact mode, moving at a linear rate of 3 to 4 mm/s to provide tissue debridement. Full-thickness articular cartilage was then harvested from each of the treatment areas. Six samples per probe were then assessed for chondrocyte viability using fluorescent double-staining followed by confocal microscopy; 6 samples per probe were assessed for metabolic activity using an 35SO4 incorporation assay; and 12 additional untreated samples were obtained to serve as controls for viability (n = 6) and metabolic activity (n = 6). RESULTS: The depth of chondrocyte death (mean +/- standard deviation) was 109.4 +/- 22.1 microm after treatment with the ACD-50 probe, and was 172.3 +/- 34.3 microm after treatment with the 2.5-mm/90 degrees probe. The 35SO4 uptake (mean +/- standard deviation) was 2584 +/- 1388 cpm/mg dry cartilage for the ACD 50 probe and 1995 +/- 852 cpm/mg of dry cartilage for the 2.5-mm/90 degrees probe. The 35SO4 uptake for the control was 2647 +/- 1380 cpm/mg dry cartilage. CONCLUSIONS: The 2 probes tested created a well-controlled debridement with smooth edges and a defined margin of chondrocyte death that extended approximately 100 to 200 microm deep to the treatment area. There does not appear to be a significant effect on the metabolic activity of the chondrocytes adjacent to the treatment zone, but with the small sample size we lacked sufficient statistical power to definitively determine these effects. CLINICAL RELEVANCE: The 2 bipolar radiofrequency probes tested created a well-controlled debridement in normal articular cartilage with smooth edges and a defined margin of chondrocyte death that extended approximately 100 to 200 microm into the treatment area. PMID- 15122141 TI - In vivo comparison of a metal versus a biodegradable suture anchor. AB - PURPOSE: This protocol compares an absorbable polylactic acid (PLA) anchor to a similarly sized metallic anchor, comparing the load to failure of the suture anchor construct in vivo in a goat model. TYPE OF STUDY: Basic science. METHODS: Thirty-nine goats underwent bilateral implantation of suture anchors at the intra articular origin of the long digital extensor tendon at the knee (stifle). Each goat randomly received an absorbable anchor in one knee and a metallic anchor in the other. At 0, 6, and 12 weeks after implantation, the suture attached to the implanted anchor was pulled to failure on a materials testing machine, annotating the load at failure and location of failure of the suture anchor construct. RESULTS: At each time period tested, no difference was found in failure load between metal and PLA suture anchor constructs, nor was there degradation in strength over time of implantation of either anchor. Most failures occurred by suture breakage, and the force required for suture breakage did not differ between anchors. Three absorbable PLA anchors failed, one (1 of 13, 7.7%) at 6 weeks, and 2 (15.4%) at 12 weeks. PLA anchor failure occurred when the suture loop eyelet pulled out of the anchor's PLA body. CONCLUSIONS: The absorbable PLA suture anchor construct, tested in an in vivo, intra-articular model, had similar strength over a 12-week period of implantation with a comparable metal anchor construct. These absorbable anchors offer a reasonable alternative to metal anchors, with comparable strength and function and advantages unique to an absorbable anchor. CLINICAL RELEVANCE: This study provides the reader with an evaluation of the in vivo function and holding strength over time of implantation of an absorbable suture anchor in comparison with a metal anchor. PMID- 15122142 TI - In vitro comparison of standard and Knotless metal suture anchors. AB - PURPOSE: Clinical experience after failed Knotless suture anchor (Mitek, Westwood, MA) fixations suggested that the Knotless anchor provides considerably less fixation stability than a standard metal anchor. The purpose of this study was to analyze soft tissue fixation to bone comparing a standard and a Knotless metal suture anchor. TYPE OF STUDY: In vitro study. METHODS: The Mitek GII and Mitek Knotless suture anchors were tested on 7 human cadaveric fresh-frozen glenoids. The anchors were inserted into the glenoid rims, and the sutures of the anchors were fixed to a metal hook attached to the cross-head of a testing machine. Cyclic loading was performed. The gap formation between the metal hook and the glenoid rim, the ultimate failure loads and the modes of failure were determined. RESULTS: The mean gap formation was significantly greater for the Knotless anchor (3.8 +/- 1.4 mm) than for the GII anchor (2.4 +/- 0.5 mm) after 25 cycles with 50 N repeated load (P =.04). The largest gap of a Knotless fixation was 5.3 mm compared with 3.0 mm for the GII. The ultimate failure load was not significantly different for the Knotless anchor (179 N) and for the GII anchor (129 N). Both anchors failed by either rupture of the suture material or by pullout of the anchors. CONCLUSIONS: The GII anchor allows significantly less displacement than the Knotless anchor. Ultimate tensile strength and mode of failure are similar. Greater displacement results in larger gap formation between the soft tissue and the bone. This might weaken and jeopardize the repair. CLINICAL RELEVANCE: If reattached soft tissues are subjected to postoperative loading, gap formation may result when using the Knotless anchor. For these conditions, suture fixation with knots may be used instead. PMID- 15122144 TI - Recurrence of synovial chondromatosis of the glenohumeral joint after arthroscopic treatment. AB - A case of primary synovial chondromatosis of the shoulder in a 15-year-old girl is presented. Plain radiographs revealed findings characteristic of synovial chondromatosis. The patient was treated by arthroscopic loose body removal and arthroscopic partial synovectomy of the glenohumeral joint. Although immediate postoperative radiographs showed no calcification in the joint, repeated radiographs at 18 months after surgery revealed recurrence of calcification in the subacromial space. Arthroscopic removal of all loose bodies and partial synovectomy appears to be a good method of giving symptomatic relief and early return to work. However, late recurrence should be anticipated. PMID- 15122143 TI - Failure of anterior shoulder instability repair caused by eyelet cutout of absorbable suture anchors. AB - Repair of soft tissue to bone is increasingly frequently performed using absorbable suture anchors. If a repair fails clinically, it is often impossible to identify the cause of failure at repeat surgery. We report on 2 cases of recurrence of instability after arthroscopic Bankart repair. In reoperation in these cases, all sutures were correctly knotted around the labrum but were intact and torn out of the anchor eyelets. No sign of anchor displacement (3 anchors in each patient) was seen. This is the first clinical report of unambiguous structural suture anchor failure. These observations emphasize the sensitivity of Bankart repair to weak links in the repair chain, which must be avoided. PMID- 15122145 TI - Femoral interference screw divergence after anterior cruciate ligament reconstruction provoking severe anterior knee pain. AB - An unusual case of anterior knee pain after anterior cruciate ligament reconstruction secondary to femoral screw divergence is described. We hypothesize that a contracture of the lateral head of the gastrocnemius, caused by irritation from the femoral screw, could increase the patellofemoral joint reaction. This would contribute to increasing the overload of the subchondral bone, which could explain the anterior knee pain. Conversely, tight gastrocnemius may lead to an increase in foot pronation of the subtalar joint, resulting in an increased valgus vector force at the knee, which can cause anterior knee pain. Finally, dorsiflexion of the talocrural joint will also decrease if the gastrocnemius is tight, provoking biomechanical limitations and possible knee problems during walking and running. PMID- 15122146 TI - Revision anterior cruciate ligament reconstruction using the regenerated semitendinosus tendon: analysis of ultrastructure of the regenerated tendon. AB - In this report, we present a case of a college skier who sustained a rerupture of the reconstructed anterior cruciate ligament (ACL) 8 months after surgery in which an autogenous semitendinosus tendon graft was used. At the revision surgery, the harvested semitendinosus tendon appeared to be regrown. Thus the regenerated tendon was reharvested, and in combination with the gracilis tendon, was used as a graft. The electron microscopic examination revealed a difference in fibril diameter between the regenerated tissue and the normal tendon. Although the regenerated semitendinosus tendon could be reharvested, the feasibility of its use for revision surgery is still to be determined. PMID- 15122147 TI - Torn discoid lateral meniscus treated using partial central meniscectomy and suture of the peripheral tear. AB - We present the cases of 5 patients with a torn discoid lateral meniscus treated using partial central meniscectomy in conjunction with the suture repair of the tear. The patients were 4 boys and 1 girl with a mean age of 15.4 years (range, 11 to 17). Preoperatively, all patients complained of knee pain during daily or sports activities. At the final follow-up evaluation, more than 2 years later, 4 patients' conditions were graded as excellent and 1 patient's condition was graded as fair according to Ikeuchi's grading scale. The average Lysholm score improved from 83.4 points (range, 70-90) to 95.8 points (range, 89-100) postoperatively. In the 3 patients who underwent second-look arthroscopy, complete healing was seen in 2 patients. One patient had severe degenerative changes in the meniscus; the repaired site was not united and required an additional partial meniscectomy along the tear. We believe that with the current advancement in arthroscopic meniscal repair techniques, a partial central meniscectomy in conjunction with the suture repair of the peripheral tear can be effective treatment for patients with a torn complete or incomplete discoid meniscus. PMID- 15122148 TI - Arthroscopic reinforced capsular shift of anterior shoulder instability. AB - This article presents an arthroscopic inferior capsular shift technique. In this technique, the same type of inferior capsular shift as with the open standard Neer procedure can be performed. After standard diagnostic shoulder arthroscopy, a bone trough is made along the capsular attachment to the humeral head using an abrader. An inverted L-shaped incision is performed in the anterior capsule. A suture is passed through the apex of this triangular flap, which is then pulled up and tied over the upper edge of the subscapularis, thus reducing the size of the wide anterior capsule. No hardware implants are used, and the procedure is not technically complicated. The surgery required fewer steps than open repair. The advantages of this technique are the preservation of the subscapularis, faster rehabilitation, and earlier return to normal activities, including sports. It also causes less postoperative range of motion limitation, while offering the same amount of capsular shift as the traditional open repair. PMID- 15122149 TI - Arthroscopic repair of SLAP lesions with a bioknotless suture anchor. AB - The diagnosis and treatment of SLAP tears have improved with the development of arthroscopic shoulder surgery techniques. With types 2 and 4 tears, the goal is to restore stability to the labrum and biceps anchor and achieve healing to the glenoid. Suture repair with anchors is currently the repair technique of choice. The purpose of this article is to report a fast and simple method for arthroscopic SLAP repair that uses knotless suture anchors and obviates complex suture management and arthroscopic knot tying. PMID- 15122150 TI - Intramedullary pressure during endoscopy of the long bone: experimental results of a new endoscopic technique. AB - This study shows the local changes in intramedullary pressure during a new endoscopic technique for the medullary canal of the long bone. The procedure of intramedullary bone endoscopy (IBE) was performed on 4 tibial amputations. By slowly pushing the endoscope distally under visual control and endoscopic preparation of the medullary canal, a "neocavum" for endoscopy was created. During the procedure, the intramedullary pressure was continuously measured: Highest peak pressure was 125 mm Hg. We therefore conclude that the procedure of IBE is a safe intervention within the medullary canal of the long bone. Local or systemic side effects, common to intramedullary reaming in fracture treatment (fat-embolism, local bone necrosis, reduction in cortical blood flow) should not be expected. PMID- 15122151 TI - The Tuckahoe knot: a secure locking slip knot. AB - We describe a simple and easy-to-tie knot that can slide and lock. The benefit of these characteristics is that when the knot is seated and locked, it is secure in that it will not slip before the 3 reversed half-hitches with alternating posts are placed. PMID- 15122152 TI - Tibial tunnel size increases after the first postoperative week. PMID- 15122154 TI - Arthroscopic excision of juxta-articular giant-cell tumor arising from the patellar tendon sheath. AB - Most giant-cell tumors of the tendon sheath are found on the flexor surface of the hand, in the fingers, wrist, or palm. The isolated discrete lesion involving tendon sheaths (localized giant-cell tumor of the tendon sheath) is rarely located intra-articularly, especially in the knee joint. To the best of my knowledge, this is the first reported case of a large, localized, juxta-articular giant-cell tumor of the patellar tendon sheath that was treated successfully with arthroscopic excision. PMID- 15122155 TI - Radial tunnel syndrome caused by ganglion cyst: treatment by arthroscopic cyst decompression. AB - Compressive neuropathies of the radial nerve at the elbow can lead to one of 2 clinical entities. Posterior interosseous syndrome is primarily a motor deficiency of the posterior interosseous nerve, and radial tunnel syndrome presents as pain along the radial tunnel and extensor muscle mass. The radial nerve can be compressed at a number of sites around the elbow. In addition, numerous mass lesions reported in the literature can cause compressive neuropathy of the radial nerve at the elbow. Standard surgical management for persistent radial tunnel syndrome that is refractory to nonsurgical treatment is open decompression of the radial nerve. Cysts occurring in other joints are commonly treated arthroscopically. Supraglenoid cysts of the shoulder, meniscal cysts in the knee, and dorsal wrist ganglia are routinely treated with arthroscopic decompression or excision with management of the underlying etiology of the cyst. We present a case of radial tunnel syndrome caused by a ganglion cyst of the proximal radioulnar joint that was treated using arthroscopic excision of the cyst and decompression of the radial nerve. PMID- 15122156 TI - Fracture of the femoral tunnel after an anterior cruciate ligament reconstruction. AB - Fractures after anterior cruciate ligament (ACL) reconstructive surgery are rare. Patella fractures can occur as a complication after bone-patellar tendon-bone autografts, and few case reports of tibia fractures have been published. Although reports of femur fractures have been published, the causes are attributed to stress risers other than the femoral tunnel. To our knowledge, this is the first case report of a femoral tunnel serving as a stress riser after an ACL reconstruction with bone-patellar tendon-bone autograft. The patient's fracture resulted from minimal trauma and required surgical fixation. PMID- 15122157 TI - Chondral injury and synovitis after arthroscopic meniscal repair using an outside in mulberry knot suture technique. AB - Arthroscopic meniscal repair has a relatively low incidence of complications. We describe a novel complication of outside-in Mulberry knot repair of the medial meniscus, namely, aseptic synovitis and chondral injury. The presence of synovitis was diagnosed clinically and by multiple arthrocenteses and confirmed, along with a chondral lesion, by repeat arthroscopy. After subtotal meniscectomy, the synovitis subsided clinically. PMID- 15122158 TI - Health care of adolescent males: overview, rationale, and recommendations. PMID- 15122159 TI - Male puberty: physical, psychological, and emotional issues. PMID- 15122160 TI - Connecting the adolescent male with health care. PMID- 15122161 TI - Providing anticipatory guidance and counseling to the adolescent male. PMID- 15122162 TI - Adolescent male sexuality. PMID- 15122163 TI - Not all adolescents are the same: addressing the unique needs of gay and bisexual male youth. PMID- 15122164 TI - Anogenital human papillomavirus infection in males. PMID- 15122165 TI - Condoms: the basics and beyond. PMID- 15122166 TI - Primary care of the elite or elite-emulating adolescent male athlete. PMID- 15122167 TI - Attention-deficit/hyperactivity disorder in adolescent males. PMID- 15122168 TI - Eating disorders in adolescent males. PMID- 15122169 TI - Boy, mediated: effects of entertainment media on adolescent male health. PMID- 15122170 TI - Genitourinary problems in adolescent males. PMID- 15122171 TI - Clinical relevance of antimicrobial resistance in the management of pneumococcal community-acquired pneumonia. AB - Streptococcus pneumoniae remains the most common bacterial cause of community acquired pneumonia, and these infections are associated with significant morbidity and mortality worldwide. A major concern is the increasing incidence of antibiotic resistance among pneumococcal isolates, which, in the case of certain of the antibiotic classes, has been associated with treatment failure. Yet despite multiple reports of infections with penicillin-resistant pneumococcal isolates, no cases of bacteriologic failure have been documented with the use of penicillin or ampicillin in the treatment of pneumonia caused by penicillin resistant pneumococci. Current prevalence and levels of penicillin resistance among pneumococal isolates in most areas of the world do not indicate a need for substantial treatment changes with regard to the use of the penicillins. For infections with penicillin-sensitive strains, penicillin or an aminopenicillin in a standard dosage will still be effective for treatment. In the cases of strains with intermediate resistance, beta-lactam agents are still considered appropriate treatment, although higher dosages are recommended. Infections with isolates of high-level penicillin resistance should be treated with alternative agents such as the third-generation cephalosporins or the new antipneumococcal fluoroquinolones. In the case of the cephalosporins, pharmacodynamic/pharmacokinetic parameters help predict which of those agents are likely to be successful, and the less active agents should not be used. Debate continues in the literature with regard to the impact of macrolide resistance on the outcome of pneumococcal pneumonia, with some investigators providing evidence of an "in vivo-in vitro paradox," referring to discordance between reported in vitro resistance and clinical success of macrolides/azalide in vivo. However, several cases of macrolide/azalide treatment failure have been documented, and many clinicians recommend that these agents not be used on their own in areas with a high prevalence and levels of macrolide/azalide resistance. However, evidence is emerging to show beneficial effects on outcome with combination therapy, especially that of a beta-lactam agent and a macrolide given together to sicker, hospitalized patients with pneumococcal pneumonia. In an attempt to prevent the emergence of resistance, it has been recommended by some that the new fluoroquinolones not be used routinely as first-line agents in the treatment of community-acquired pneumonia; instead, they say, these agents should be reserved for patients who are allergic to the commonly used beta-lactam agents, for infections known to be or suspected of being caused by highly resistant strains, and for patients in whom initial therapy has failed. PMID- 15122172 TI - Mechanisms of and perspectives on the mesenchymal stem cell in immunotherapy. AB - Mesenchymal stem cells (MSCs) are an important cell population in the bone-marrow microenvironment and are considered to be engaged mainly in the support of hematopoiesis. Recent work has shown that MSCs also have profound immunomodulatory function, both in vitro and in vivo. Because MSCs can be expanded rapidly to the numbers required for clinical application, several preclinical and clinical studies have been performed in the areas of immune diseases and bone-marrow transplantation. In this review we discuss the mechanisms underlying the MSC's immunomodulating properties and its potential applications. PMID- 15122173 TI - Attenuation of the alcohol preference of C57BL/6 mice during chronic renal failure. AB - The C57BL/6 inbred mouse strain is known for its strong, genetically determined preference for alcohol over water. In this study we examined the voluntary alcohol consumption (VAC) of C57BL/6 mice during chronic renal failure (CRF). Two weeks after the surgical induction of renal failure, CRF mice, together with normal and sham-operated control mice, were submitted to a standard 24-day VAC protocol. The mice were offered water for the first 6 days (period of acclimatization), alcohol (10% ethanol solution) for the next 4 days (period of forced alcohol exposure), and a choice between water and alcohol for the last 14 days (VAC period). The results (mean +/- SEM) obtained from the last 8 days of the VAC period were significantly different (P <.05) between CRF mice and the 2 control groups. As expected, CRF mice had a higher total fluid intake than did normal and sham-operated controls (9.5 +/- 0.2 vs 5.4 +/- 0.2 and 5.4 +/- 0.2 g/d). Surprisingly, despite their increased total fluid consumption, CRF mice nearly abolished their absolute alcohol intake compared with that of both control groups (3.2 +/- 0.5 vs 13.1 +/- 0.8 and 14.2 +/- 1.1 g alcohol/kg body wt/d). The resulting alcohol preference ratio (g alcohol/g total fluid) was markedly decreased in the CRF mice compared with that in both control groups (0.09 +/- 0.01 vs 0.62 +/- 0.03 and 0.64 +/- 0.05). We conclude that the innate alcohol preference of C57BL/6 mice is nearly abolished during CRF. Additional studies to clarify the mechanism of this striking change in drinking pattern are required, with special emphasis on the possible role of angiotensin II, which is involved in thirst regulation and known to reduce the alcohol consumption of normal alcohol-preferring rats. PMID- 15122174 TI - A novel modification of the Thrombelastograph assay, isolating platelet function, correlates with optical platelet aggregation. AB - Flow cytometry, singlet platelet counting, and optical aggregation have been used to monitor clopidogrel and glycoprotein IIb/IIIa (GPIIb/IIIa) platelet antagonists. Optical aggregation is considered the gold standard, but neither it nor flow cytometry is convenient in larger-scale clinical studies or point-of care systems. Singlet platelet counting, a point-of-care assay correlated with optical platelet aggregation, only provides a measurement of platelet function at a single point in time. The Thrombelastograph is used to assay whole blood for thrombin-generated maximal clot-shear elasticity, referred to as the maximal amplitude (MA). Although platelet dysfunction, thrombocytopenia, and the in vitro effect of strong inhibitors such as IIb/IIIa antagonists can be observed, with thrombin generation milder platelet inhibitors cannot be assessed. We modified the Thromboelastograph assay, using reptilase and factor XIIIa, to form a clot, without thrombin generation, in heparinized whole blood. The resulting clot MA is dependent on added platelet agonists such as ADP or arachidonic acid, is sensitive to platelet antagonists, and provides a continuous measure of platelet function more analogous and better correlated with optical aggregation. This novel modification of the Thromboelastograph assay should prove to be a useful point-of-care whole-blood assay with which to monitor the effects of GPIIb/IIIa, ADP, and thromboxane A(2)-receptor-inhibiting drugs in patients. PMID- 15122175 TI - Lack of good correlation of serum CC-chemokine levels with human immunodeficiency virus-1 disease stage and response to treatment. AB - Three CC-chemokines-MIP-1alpha (CCL3), MIP-1beta (CCL4), and RANTES (CCL5)-are natural ligands for the human immunodeficiency virus-1 (HIV-1) coreceptor CCR5. To determine correlations between CC-chemokines and HIV-1 disease stage or response to treatment, we examined serum levels of MIP-1alpha, MIP-1beta, and RANTES in 60 infected patients during 18 months while they were taking highly active antiretroviral therapy (HAART). Our results demonstrate that serum levels of MIP-1alpha and RANTES were increased in HIV-1-infected individuals compared with those in healthy controls. We found no significant differences among 4 clinical stages of HIV-1 infection in the serum levels of three CC-chemokines. Longitudinal HAART analyses revealed a pronounced decline in serum MIP-1alpha levels over time. We found no difference in this decline between HAART responders and nonresponders. These findings indicate that production of MIP-1alpha and RANTES changes during HIV-1 infection and treatment; however, our results suggest that serum levels of CC-chemokines should not be used as biomarkers for HIV-1 disease stage or response to treatment. PMID- 15122176 TI - Glycoprotein IIb/IIIa inhibitors increase COAT-platelet production in vitro. AB - Platelets activated simultaneously with thrombin and collagen reveal a subpopulation of cells that express on their surfaces high levels of several alpha-granule proteins, including factor V and fibrinogen; these COAT platelets (collagen and thrombin-activated platelets) represent roughly 30% of the total population. Evidence of enhanced stability of proteins on the COAT-platelet surface was provided by the observation that PAC-1, a mAB recognizing the activated form of glycoprotein (GP) IIb/IIIa, did not inhibit fibrinogen binding to COAT-platelets. We therefore undertook a systematic evaluation of the effects of other GP IIb/IIIa inhibitors on the production of COAT platelets. Not only did GP IIb/IIIa antagonists fail to inhibit the retention of fibrinogen on COAT platelets, but several actually increased the absolute percentage of COAT platelets produced. The increases over control values in the presence of eptifibatide, tirofiban, and DMP-802 were 1.36-, 1.20-, and 1.05-fold, respectively (P <.01 for each comparison). COAT-platelet production in the presence of abciximab was not significantly affected. However, platelet activation with thrombin plus ALB6, an Fc-receptor agonist, produces a product, referred to as FcRT platelets, that is indistinguishable from COAT platelets; all 4 GP IIb/IIIa antagonists tested potentiated formation of FcRT platelets. These findings indicate that fibrinogen binding to COAT platelets and FcRT platelets is not affected by available GP IIb/IIIa inhibitors. More importantly, our study demonstrates a potentiation of COAT-platelet production by some GP IIb/IIIa antagonists that may be relevant to the observation that long-term administration of orally available GP IIb/IIIa inhibitors not only failed to protect patients but actually increased the frequency of acute coronary events. PMID- 15122177 TI - Hand-carried cardiac ultrasound as a tool to screen for important cardiovascular disease in an underserved minority health care clinic. AB - The disparity in cardiovascular outcomes among racial and social strata may be, in part, because of delayed detection of cardiovascular disease in minority patients. The low cost and portability of hand-carried cardiac ultrasound devices may make screening of underserved patients for cardiac disease feasible. A general internist evaluated 153 patients at a clinic serving an underserved population with a hand-carried cardiac ultrasound device. A total of 27 cases of significant valvular heart disease or ventricular dysfunction were detected in 19 patients (12.4%). Detection of a major cardiac abnormality could not be predicted by cardiac risk factors, age, or chief symptom, whereas patients presenting for new or acute clinic visits were more likely to have an abnormality. The low cost and portability of hand-carried cardiac ultrasound devices may make them important tools for the early detection of cardiovascular disease in minority and underserved populations and, thereby, help to reduce disparities in cardiovascular outcomes. PMID- 15122178 TI - Monitoring change in the three-dimensional shape of the human left ventricle. AB - BACKGROUND: Characterizing left ventricular (LV) remodeling after myocardial infarction or LV shape change resulting from LV shape-restoration operation can yield valuable prognostic information. However, current methods measure only global parameters of LV shape. METHODS: We developed and validated a method for measuring change in regional LV shape by aligning a patient's follow-up 3 dimensional LV surface reconstruction to baseline surface. We tested the diagnostic power of 6 distance functions to detect a known shape deformation. To create the test data, the LV endocardial surface of a control subject was reconstructed using 3-dimensional echocardiographic techniques. The surface was deformed 9 different ways to model LV dilation (3 different locations and severities). Normal shape variability was defined from 18 serial studies of 6 control subjects. The severity of regional dilation was computed as the orthogonal distance between the aligned baseline and deformed LV surfaces. Deformation was quantified according to regional location using the 16-segment map of the LV. RESULTS: Normal LV shape variability was 3.38 mm. The LV deformations ranged from 2.95 to 8.02 mm. Gaussian distance function produced the highest accuracy for measuring deformation distances (P <.005 by analysis of variance). In addition, the gaussian function correctly identified the location of the maximum deformation in 6 of the 9 distorted surfaces. In the 3 remaining surfaces, the gaussian alignment selected an adjacent basal segment with a similar deformation distance (mean error: 0.2 +/- 0.17 mm). The gaussian function's accuracy in pinpointing the deformation equaled or exceeded the performance of the other 5 functions tested. CONCLUSION: This new method of aligning 3-dimensional LV surfaces in space facilitates detecting, measuring, and localizing regional shape change in the human LV independent of anatomic landmarks or geometric references. Potential applications include quantitative monitoring of change in regional LV shape after a pathologic process and/or surgical procedure to document efficacy of treatment and to assess prognosis. PMID- 15122179 TI - Echocardiographic assessment of regional ventricular function after device-based change of left ventricular shape. AB - We assessed the effects of implantation of Myosplint (Myocor, Maple Grove, Minn), a device that changes left ventricular (LV) cross-sectional shape from circular to bilobar, on regional LV function. A total of 10 open-chest dogs with tachycardia-induced cardiomyopathy were studied before and after Myosplint implantation. LV cross-sectional epicardial echocardiography at the papillary muscle level was performed along with acquisition of hemodynamic data. LV normalized thickening, fractional thickening, end-diastolic thickness, and end diastolic curvatures were calculated for 10 LV segments. Myosplint implantation did not affect LV hemodynamics, but decreased average end-diastolic curvature (P <.0001) and increased its segmental heterogeneity (P <.0001). There was no change in average fractional thickening, whereas normalized thickening increased (P =.05). In contrast, segmental heterogeneity of both normalized and fractional thickening increased (P =.02 and P =.01, respectively). Structural modeling confirmed that Myosplint implantation increases regional stress heterogeneity and curvature heterogeneity. LV cross-sectional shape markedly affects regional LV performance. PMID- 15122180 TI - Midwall mechanics in physiologic and hypertensive concentric hypertrophy. AB - OBJECTIVE: We sought to analyze and compare midwall fractional shortening (mFS), and its relations with circumferential end-systolic stress (cESS) and relative wall thickness (RWT), among subjects with physiologic concentric left ventricular (LV) hypertrophy, patients with hypertension and concentric LV hypertrophy, and control subjects. METHODS: A total of 51 normotensive athletes and 56 young patients with hypertension and echocardiographic evidence of concentric LV hypertrophy were enrolled. In addition, 49 age- and sex-matched control subjects were recruited. LV cavity size and wall thicknesses, LV mass, RWT, cESS, and mFS were determined by echocardiography. RESULTS: The 3 groups were similar in age, sex, height, weight, body surface area, LV diameters, and conventional indices of systolic function. LV thicknesses, RWT, LV mass, and LV mass index were similarly increased in the athletes and in the hypertensive group when compared with the control subjects. A similar depression in mFS was observed in both the athletes (22.4 +/- 2.6%) and hypertensive group (22.5 +/- 3.6%) in comparison with the control group (24.5 +/- 2.5%, P =.0003). The depression in mFS was still significant after taking into account the effect of cESS, but was no more evident after indexation of mFS by RWT or LV mass. At regression analysis, the relation between mFS and cESS showed a steeper negative slope in the patients with hypertension than in the other 2 groups. CONCLUSIONS: MFS is similarly depressed in physiologic and hypertensive concentric LV hypertrophy. The depression is abolished by adjustment for RWT or LV mass, suggesting that geometric factors are the major determinants of midwall performance in both types of concentric LV hypertrophy. However, an impaired response to different values of cESS seems to exist only in hypertensive concentric LV hypertrophy, because increasing levels of cESS may be associated with more evident mFS depression in patients with hypertension than in the athletes and control subjects. PMID- 15122181 TI - Triphasic mitral inflow velocity with middiastolic filling: clinical implications and associated echocardiographic findings. AB - To explore underlying mechanisms and clinical implications of middiastolic filling, we measured early and late mitral inflow velocities, deceleration time of early mitral inflow velocity, and early diastolic mitral annular velocity (E') recorded by pulsed wave Doppler echocardiography in 3 cardiac cycles of 35 patients with prominent mitral inflow (middiastolic flow velocity > or = 0.2 m/s). E' was measured at the septal corner of the mitral annulus by Doppler tissue echocardiography from the apical 4-chamber view and was found to be reduced (E' < 0.1 m/s) in all patients; early mitral inflow velocity/E' ratio was > 10 in all but 1 patient. Valsalva maneuver unmasked delayed relaxation in 15 (88%) of 17 patients and abolished middiastolic filling in 10 (59%). Triphasic mitral inflow with middiastolic flow is related to elevated filling pressure, delayed myocardial relaxation, and slow heart rate, indicating advanced diastolic dysfunction. PMID- 15122182 TI - Preload-dependent variation of the propagation velocity in patients with congestive heart failure. AB - BACKGROUND: Although color Doppler M-mode propagation velocity (Vp) is preload independent, the variation in Vp with the temporal variation of preload in the clinical setting has not been evaluated. Because left ventricular filling pressure changes dramatically with treatment of congestive heart failure (CHF), we hypothesized that preload-dependent variations in Vp occur with treatment of CHF. METHODS: We performed Doppler echocardiographic and hemodynamic evaluation in 24 patients with CHF (15 men, 62 +/- 10 years) at initial presentation (baseline study) and after CHF had improved with therapy (second study). RESULTS: The interval between the baseline and the second study was 48.6 +/- 21.5 hours. Vp decreased between the baseline study (41 +/- 5 cm/s) and the second study (28 +/- 5 cm/s, P <.0001). Only the change in pulmonary capillary wedge pressure ( 8.3 +/- 3.3 mm Hg) between the baseline and second study was an independent predictor of the change in Vp (-12.5 +/- 5.9 cm/s) by stepwise linear regression (r = 0.68, P =.0002). CONCLUSIONS: Vp decreases significantly with decreases in pulmonary capillary wedge pressure with the treatment of CHF. The preload dependent variation should be taken into account in the assessment of Vp in patients with CHF. PMID- 15122183 TI - Myocardial performance index in pediatric patients after cardiac transplantation. AB - BACKGROUND: Myocardial performance index (MPI) has been shown to be a reliable indicator of ventricular performance. This study determined MPI values in pediatric patients after cardiac transplantation without endomyocardial rejection. METHODS: MPI was determined in 41 pediatric patients after cardiac transplantation, without evidence of microscopic rejection, and in 31 pediatric control subjects. RESULTS: MPI in the transplantation group (0.41 +/- 0.12) was higher than in the control group (0.31 +/- 0.09; P =.0003). Isovolumic relaxation time and isovolumic relaxation time/ejection time were higher in the transplant group (55 +/- 20 milliseconds and 0.22 +/- 0.07, respectively) compared with the control group (41 +/- 10 milliseconds and 0.16 +/- 0.06, respectively; P =.0002). Isovolumic contraction time and isovolumic contraction time/ejection time were similar in the transplant group (48 +/- 23 milliseconds and 0.19 +/- 0.09, respectively) and control group (43 +/- 21 milliseconds and 0.16 +/- 0.08, respectively; P = not significant). CONCLUSIONS: Pediatric patients after cardiac transplantation without endomyocardial rejection have a higher MPI compared with a normative pediatric control population. The difference appears to be related to abnormal diastolic function. PMID- 15122185 TI - Is quantitative interpretation likely to increase sensitivity of dobutamine stress echocardiography? A study of false-negative results. AB - BACKGROUND: False-negative interpretations of dobutamine stress echocardiography (DSE) may be associated with reduced wall stress. Using measurements of contraction, we sought whether these segments were actually ischemic but unrecognized or showed normal contraction. METHODS: We studied 48 patients (29 men; mean age 60 +/- 10 years) with normal regional function on the basis of standard qualitative interpretation of DSE. At coronary angiography within 6 months of DSE, 32 were identified as having true-negative and 16 as having false negative results of DSE. Three apical views were used to measure regional function with color Doppler tissue, integrated backscatter, and strain rate imaging. Cyclic variation of integrated backscatter was measured in 16 segments, and strain rate and peak systolic strain was calculated in 6 walls at rest and peak stress. RESULTS: Segments with false-negative results of DSE were divided into 2 groups with and without low wall stress according to previously published cut-off values. Age, sex, left ventricular mass, left ventricular geometric pattern, and peak workload were not significantly different between patients with true- and false-negative results of DSE. Importantly, no significant differences in cyclic variation and strain parameters at rest and peak stress were found among segments with true- and false-negative results of DSE with and without low wall stress. Stenosis severity had no influence on cyclic variation and strain parameters at peak stress. CONCLUSIONS: False-negative results of DSE reflect lack of ischemia rather than underinterpretation of regional left ventricular function. Quantitative markers are unlikely to increase the sensitivity of DSE. PMID- 15122184 TI - The relation between quantitative right ventricular ejection fraction and indices of tricuspid annular motion and myocardial performance. AB - BACKGROUND: Assessment of right ventricular (RV) function remains challenging because of complex RV chamber geometry and a paucity of easily derived and objective functional methods. METHODS: Visual 2-dimensional echocardiographic estimates of RV ejection fraction (EF), tricuspid annular plane systolic excursion, Doppler tissue imaging, and myocardial performance index (MPI) were compared with biplanar Simpson's rule RV EF in 101 consecutive patients. Data were analyzed using simple linear regression and receiver operating characteristic curves. RESULTS: RV EF was significantly correlated with tricuspid annular plane systolic excursion (r = 0.48, P <.0001), Doppler tissue imaging peak systolic velocity (r = 0.45, P <.0001), and MPI (r = -0.38, P =.006). Using a Simpson's RV EF < 50%, the sensitivity, specificity, and positive and negative predictive values of tricuspid annular plane systolic excursion < 1.5 cm were 59%, 94%, 71%, and 89%; of Doppler tissue imaging peak systolic velocity < 10 cm/s were 59%, 92%, 67%, and 89%; and of MPI < 0.40 were 100%, 35%, 29%, and 100%, respectively. The area under the receiver operating characteristic curves was similar for the 3 indices. CONCLUSIONS: Measurements of tricuspid annular motion are easy to obtain, correlate with Simpson's RV EF, and have a high specificity and negative predictive value for detecting abnormal RV systolic function; and the MPI, although not specific, has high sensitivity and negative predictive value for detecting abnormal RV systolic function. PMID- 15122186 TI - Accuracy of coronary artery anatomy using two-dimensional echocardiography in d transposition of great arteries using a two-reviewer method. AB - We evaluated echocardiographic accuracy for defining coronary artery course in d transposition of great arteries and the impact of a 2-reviewer method on this accuracy. The echocardiogram reports of 108 patients with d-transposition of great arteries were reviewed for coronary anatomy and compared with the operative report. In method 1, from January 1995 to December 1997, a single reader performed the echocardiogram. In method 2, from January 1998 to December 2000, 2 readers scanned individually and a consensus diagnosis was made. Comparing methods 1 and 2, the sensitivity of the echocardiogram to detect variants in coronary anatomy was 68% versus 86%, and negative predictive value was 72% versus 91%. Using a 2-reviewer method improved the accuracy of echocardiographic diagnosis of coronary anatomy with d-transposition of great arteries, aiding in risk assessment and treatment of the patient preoperatively. This study also illustrates that echocardiographic accuracy may be lower in an institution with a surgical volume more representative of the usual pediatric cardiothoracic surgical center. PMID- 15122187 TI - Diastolic right-to-left shunting in a patient with atrial septal defect and pericardial tamponade. AB - We present the case of a patient with an atrial septal defect who presented with hypotension, hypoxia, and cyanosis as a result of pericardial tamponade. The classic findings of pulsus paradoxus and low measured right heart output were not present. Echocardiography demonstrated the atrial septal defect, pericardial effusion, and diastolic chamber collapse. Furthermore, Doppler study showed not only a large left-to-right shunt but also a diastolic right-to-left shunt, which explained the patient's hypoxia. PMID- 15122188 TI - Diagnoses of right-sided empyema complicating tricuspid valve endocarditis during transesophageal echocardiography. AB - A 25-year-old man, who was a known intravenous drug user, underwent transesophageal echocardiography as part of investigation for suggested infective endocarditis. Tricuspid valve infective endocarditis was diagnosed. The simultaneous visualization of right-sided empyema, previously undiagnosed, by transesophageal echocardiography, led to operative treatment by thoracoscopy and right lung decortication. PMID- 15122189 TI - Coronary collaterals by transthoracic echocardiography in coronary artery disease. AB - Demonstration of coronary collaterals has been mostly done by coronary angiographic techniques. Coronary collaterals have been demonstrated by transthoracic echocardiography for patients with anomalous origin of coronary arteries from pulmonary artery. Indirect assessment of collaterals to an infarct related artery has been done using myocardial contrast echocardiography. We describe in this article 3 patients with coronary artery disease in whom coronary collaterals were demonstrated by transthoracic echocardiography before angiography. To our knowledge, coronary collateral demonstration by transthoracic echocardiography before angiography in patients with coronary artery disease has not been previously reported. PMID- 15122190 TI - Echocardiographic approach in a new left ventricular assist device: Impella Recover 100. AB - Echocardiographic evaluation represents a basic tool for the treatment of patients undergoing ventricular assist device implant. Impella Recover 100 is a new left intraventricular assist device proposed for short-term mechanical circulatory support to be implanted without cardiopulmonary bypass. We report our experience with echocardiographic monitoring on a patient with ischemic cardiomyopathy who underwent Impella Recover 100 implant as a bridge to heart transplant. During the surgical procedure echocardiography had an essential role for anatomic evaluation of the heart and correct intraventricular positioning of the device. Moreover, during the whole period of assistance, echocardiography contributed to assessment of patient-device interaction and to check the device when malfunction was suggested. PMID- 15122192 TI - Acquired partially flail leaflet causing severe mitral regurgitation in a congenital double-orifice mitral valve. AB - Double orifice mitral valve is a rare congenital malformation. We report a 48 year-old woman with new onset congestive heart failure and mitral regurgitation. Transesophageal echocardiography showed a partially flail mitral valve and severe mitral regurgitation. A deep transgastric view showed a double orifice mitral valve with a smaller accessory anterolateral orifice, and identified the origin of the regurgitant jet from the larger posteromedial orifice. PMID- 15122191 TI - Echocardiographic features of an unruptured mycotic aneurysm of the right aortic sinus of Valsalva. AB - This report describes a 25-year-old man with a pierced tongue in whom Streptococcus constellatus endocarditis of the aortic valve developed. Bacterial endocarditis in this patient was complicated by the development of a mycotic aneurysm of the right aortic sinus of Valsalva. Transthoracic and transesophageal echocardiographic studies were useful for the diagnosis of this rare lesion. Findings were confirmed at operation. PMID- 15122193 TI - Blood cyst of the mitral valve apparatus in a woman with a history of orthotopic liver transplantation. AB - Blood cysts within the heart are rare findings in adults. We describe a case of a woman with a history of orthotopic liver transplantation who presented for an echocardiogram to evaluate a potential source of cerebrovascular embolism. A cystic mass attached to the submitral valve apparatus was identified by transthoracic and confirmed by transesophageal echocardiography. Surgical exploration and pathologic examination confirmed the mass to be a blood cyst attached to the chordal apparatus of the posterior papillary muscle. This case report highlights the use of echocardiography in diagnosing intracardiac blood cysts. PMID- 15122194 TI - Fluoroscopic balloon-guided transesophageal echocardiography in a patient with Zenker's diverticulum. AB - During the past 20 years, transesophageal echocardiography (TEE) became an important diagnostic technique. Indications for TEE include: defining the cause and severity of native valve disease, particularly mitral regurgitation; detecting vegetations and other sequelae of endocarditis; assessing prosthetic valve function; and identifying a potential cardiac source for emboli.(1) TEE is usually well tolerated and is associated with few adverse events. However, structural abnormalities of the esophagus such as diverticula, stenoses, tumors, and advanced varices are relative contraindications to TEE because of the technical difficulties associated with probe advancement and the risk of esophageal perforation.(2) This report describes the successful performance of TEE in a patient with a Zenker's diverticulum. The patient was severely symptomatic of atrial fibrillation and was a poor candidate for long-term anticoagulation. Therefore, it was necessary to rule out a thrombus before cardioversion. Because the Zenker's diverticulum was large, a novel approach was taken using a balloon to occlude the orifice allowing safe passage of the TEE probe. PMID- 15122195 TI - Transient ischemic attack in a patient with pulmonary vein thrombosis after left upper lobectomy for squamous cell lung cancer. PMID- 15122196 TI - Retained left atrial catheter after coronary artery bypass operation. PMID- 15122197 TI - Pulmonary vein stenosis after radiofrequency pulmonary vein isolation for atrial fibrillation. PMID- 15122198 TI - Selectins in T-cell recruitment to non-lymphoid tissues and sites of inflammation. PMID- 15122199 TI - Co-inhibitory molecules of the B7-CD28 family in the control of T-cell immunity. PMID- 15122200 TI - CARMA1, BCL-10 and MALT1 in lymphocyte development and activation. PMID- 15122201 TI - Lymphocyte trafficking across high endothelial venules: dogmas and enigmas. PMID- 15122202 TI - The weight of leptin in immunity. AB - Leptin is an adipocyte-derived hormone/cytokine that links nutritional status with neuroendocrine and immune functions. As a hormone, leptin regulates food intake and basal metabolism, and is sexually dimorphic - that is, its serum concentration is higher in females than in males with a similar body fat mass. As a cytokine, leptin can affect thymic homeostasis and the secretion of acute-phase reactants such as interleukin-1 and tumour-necrosis factor. Similar to other pro inflammatory cytokines, leptin promotes T helper 1 (TH1)-cell differentiation and can modulate the onset and progression of autoimmune responses in several animal models of disease. Here, we review the advances and controversy for a role of leptin in the pathophysiology of immune responses. PMID- 15122203 TI - Novel ChIP-based strategies to uncover transcription factor target genes in the immune system. PMID- 15122204 TI - Concomitant regulation of T-cell activation and homeostasis. PMID- 15122205 TI - The TOR pathway: a target for cancer therapy. PMID- 15122206 TI - The proteasome: a suitable antineoplastic target. PMID- 15122207 TI - The discovery of receptor tyrosine kinases: targets for cancer therapy. PMID- 15122208 TI - Molecules and mechanisms of the graft-versus-leukaemia effect. PMID- 15122209 TI - Searching for the hereditary causes of renal-cell carcinoma. PMID- 15122210 TI - C/EBPalpha mutations in acute myeloid leukaemias. PMID- 15122211 TI - The promise of cancer vaccines. PMID- 15122212 TI - Why French politics and science don't mix. PMID- 15122213 TI - Towards biomimetic architecture. PMID- 15122215 TI - Nanoparticles: spontaneous ligand organization. PMID- 15122216 TI - Microfluidics: a few good tricks. PMID- 15122217 TI - Molecular filters: pores within pores. PMID- 15122218 TI - Molecular electronics: back under control. PMID- 15122221 TI - Synthesis of branched 'nanotrees' by controlled seeding of multiple branching events. AB - The formation of nanostructures with controlled size and morphology has been the focus of intensive research in recent years. Such nanostructures are important in the development of nanoscale devices and in the exploitation of the properties of nanomaterials. Here we show how tree-like nanostructures ('nanotrees') can be formed in a highly controlled way. The process involves the self-assembled growth of semiconductor nanowires via the vapour-liquid-solid growth mode. This bottom up method uses initial seeding by catalytic nanoparticles to form the trunk, followed by the sequential seeding of branching structures. Each level of branching is controlled in terms of branch length, diameter and number, as well as chemical composition. We show, by high-resolution transmission electron microscopy, that the branching mechanism gives continuous crystalline (monolithic) structures throughout the extended and complex tree-like structures. The controlled seeding method that we report here has potential as a generic means of forming complex branching structures, and may also offer opportunities for applications, such as the mimicking of photosynthesis in nanotrees. PMID- 15122222 TI - Days of molecular medicine 2004. PMID- 15122223 TI - Antidepressant reputation falls to new lows. PMID- 15122225 TI - Outsourcing clinical trials to India rash and risky, critics warn. PMID- 15122228 TI - Debate escalates on source of sub-Saharan Africa's AIDS epidemic. PMID- 15122229 TI - Despite hype, not all statins are the same, experts say. PMID- 15122233 TI - The heart of the matter. PMID- 15122234 TI - Nasal colonization by Staphylococcus aureus. PMID- 15122238 TI - Tanning before transplant: lancing the Langerhans cell. PMID- 15122239 TI - Finally, mice with CF lung disease. PMID- 15122240 TI - Fat hormones pull their weight in the CNS. PMID- 15122241 TI - The mosquito's innate sting. PMID- 15122242 TI - A COG in the sugar machine. PMID- 15122243 TI - Arming the osteoclast. PMID- 15122244 TI - Tangling with hypothermia. PMID- 15122246 TI - Cardiac channelopathies: it's in the genes. PMID- 15122248 TI - A decade of discoveries in cardiac biology. AB - The heart is the first organ to form in the embryo, and all subsequent events in the life of the organism depend on its function. Inherited mutations in cardiac regulatory genes give rise to congenital heart disease, the most common form of human birth defects, and abnormalities of the adult heart represent the most prevalent cause of morbidity and mortality in the industrialized world. The past decade has marked a transition from physiological and functional studies of the heart toward a deeper understanding of cardiac function (and dysfunction) at genetic and molecular levels. These discoveries have provided new therapeutic approaches for prevention and palliation of cardiac disease and have raised new questions, challenges and opportunities for the future. PMID- 15122249 TI - Dendritic cell immunotherapy: mapping the way. AB - Dendritic cells (DCs) are the professional antigen-presenting cells of the immune system, with the potential to either stimulate or inhibit immune responses. Exploiting the immune-regulatory capacities of dendritic cells holds great promise for the treatment of cancer, autoimmune diseases and the prevention of transplant rejection. Although early clinical trials indicate that DC vaccines can induce immune responses in some cancer patients, careful study design and use of standardized clinical and immunological criteria are needed. PMID- 15122251 TI - Bioluminescent imaging of Cdk2 inhibition in vivo. AB - Many proteins and pathways of pharmaceutical interest impinge on ubiquitin ligases or their substrates. The cyclin-dependent kinase (Cdk) inhibitor p27, for example, is polyubiquitylated in a cell cycle-dependent manner by a ubiquitin ligase complex containing the F-box protein Skp2. Regulated turnover of p27 is due, at least partly, to its phosphorylation by Cdk2 on threonine 187, which generates a Skp2-binding site. We made a p27-luciferase (p27Luc) fusion protein and show here that its abundance, like that of p27, is regulated by Skp2 in a cell cycle-dependent manner. As predicted, p27Luc levels increased after blocking Cdk2 activity with inhibitory proteins, peptides or small interfering RNA (siRNA). Accumulation of p27Luc in response to Cdk2 inhibitory drugs (flavopiridol and R-roscovitine) was demonstrable in human tumor cells in vivo using noninvasive bioluminescent imaging. In theory, the approach described here could be used to develop bioluminescent reporters for any drug target that directly or indirectly affects the turnover of a ubiquitin ligase substrate. PMID- 15122252 TI - Antigen-induced tolerance by intrathymic modulation of self-recognizing inhibitory receptors. AB - CD1d-restricted invariant natural killer T cell (iNKT cells) have a limited T cell receptor (TCR) repertoire and share characteristics common to T cells and natural killer cells. While intrathymic selection facilitates the production of T cells carrying self major histocompatibility complex-restricted TCRs, natural killer cells carry an appropriate repertoire of self major histocompatibility complex-recognizing receptors to avoid self-reactivity. Here we show that chronic exposure to specific glycolipid antigen resulted in iNKT cell disappearance and thymus-dependent repopulation of iNKT cells with increased expression of inhibitory Ly-49 molecules that resulted in impaired responsiveness. Thymic selection of peripheral Ly-49-expressing iNKT cell repertoire inhibited cytokine production and other functions in vivo. These observations emphasize the acquisition of self-recognizing inhibitory receptors on NKT cells as a previously unknown mechanism of thymic tolerance after chronic antigen exposure. PMID- 15122253 TI - Hot-spot residue in small heat-shock protein 22 causes distal motor neuropathy. AB - Distal hereditary motor neuropathies are pure motor disorders of the peripheral nervous system resulting in severe atrophy and wasting of distal limb muscles. In two pedigrees with distal hereditary motor neuropathy type II linked to chromosome 12q24.3, we identified the same mutation (K141N) in small heat-shock 22-kDa protein 8 (encoded by HSPB8; also called HSP22). We found a second mutation (K141E) in two smaller families. Both mutations target the same amino acid, which is essential to the structural and functional integrity of the small heat-shock protein alphaA-crystallin. This positively charged residue, when mutated in other small heat-shock proteins, results in various human disorders. Coimmunoprecipitation experiments showed greater binding of both HSPB8 mutants to the interacting partner HSPB1. Expression of mutant HSPB8 in cultured cells promoted formation of intracellular aggregates. Our findings provide further evidence that mutations in heat-shock proteins have an important role in neurodegenerative disorders. PMID- 15122254 TI - Mutant small heat-shock protein 27 causes axonal Charcot-Marie-Tooth disease and distal hereditary motor neuropathy. AB - Charcot-Marie-Tooth disease (CMT) is the most common inherited neuromuscular disease and is characterized by considerable clinical and genetic heterogeneity. We previously reported a Russian family with autosomal dominant axonal CMT and assigned the locus underlying the disease (CMT2F; OMIM 606595) to chromosome 7q11 q21 (ref. 2). Here we report a missense mutation in the gene encoding 27-kDa small heat-shock protein B1 (HSPB1, also called HSP27) that segregates in the family with CMT2F. Screening for mutations in HSPB1 in 301 individuals with CMT and 115 individuals with distal hereditary motor neuropathies (distal HMNs) confirmed the previously observed mutation and identified four additional missense mutations. We observed the additional HSPB1 mutations in four families with distal HMN and in one individual with CMT neuropathy. Four mutations are located in the Hsp20-alpha-crystallin domain, and one mutation is in the C terminal part of the HSP27 protein. Neuronal cells transfected with mutated HSPB1 were less viable than cells expressing the wild-type protein. Cotransfection of neurofilament light chain (NEFL) and mutant HSPB1 resulted in altered neurofilament assembly in cells devoid of cytoplasmic intermediate filaments. PMID- 15122255 TI - Duplicate genes increase gene expression diversity within and between species. AB - Using microarray gene expression data from several Drosophila species and strains, we show that duplicated genes, compared with single-copy genes, significantly increase gene expression diversity during development. We show further that duplicate genes tend to cause expression divergences between Drosophila species (or strains) to evolve faster than do single-copy genes. This conclusion is also supported by data from different yeast strains. PMID- 15122257 TI - Dynein motors transport activated Trks to promote survival of target-dependent neurons. AB - Mutations that alter dynein function are associated with neurodegenerative diseases, but it is not known why defects in dynein-dependent transport impair neuronal survival. Here we show that dynein function in axons is selectively required for the survival of neurons that depend on target-derived neurotrophins. Stimulation of axon terminals with neurotrophins causes internalization of neurotrophin receptors (Trks). Using real-time imaging of fluorescently tagged Trks, we show that dynein is required for rapid transport of internalized, activated receptors from axon terminals to remote cell bodies. When dynein-based transport is inhibited, neurotrophin stimulation of axon terminals does not support survival. These studies indicate that defects in dynein-based transport reduce trafficking of activated Trks and thereby obstruct the prosurvival effect of target-derived trophic factors, leading to degeneration of target-dependent neurons. PMID- 15122256 TI - Long lifespan in worms with long telomeric DNA. AB - Telomere length is a crucial factor in senescence, but it has not been determined whether animals with long telomeres live longer than those with normal-length telomeres in the isogenic background of a given species. Here we show the effect of long telomeres on lifespan in the nematode Caenorhabditis elegans. We examined the effect of telomere length on lifespan by overexpressing HRP-1, a telomere binding protein, which gradually increased telomere length in worms. Worms with longer telomeres lived longer. We confirmed that the extension of lifespan was due to the increased telomere length, and not to the overexpression of HRP-1 per se, by examining the lifespans of nontransgenic progeny of the transgenic worms, who retained the longer telomeres. The lifespan-extending effect of long telomeres was dependent on daf-16. The number of germ stem cells was not affected in worms with long telomeres, indicating that the telomere effect on lifespan is independent of germ stem cell cycling. Worms with long telomeres were more resistant to heat stress. Taken together, our results suggest that signaling may be initiated in postmitotic somatic cells by telomere length to regulate organismal lifespan. PMID- 15122258 TI - Sliding across a cytoskeletal milestone. PMID- 15122260 TI - N-terminal acetylation targets GTPases to membranes. PMID- 15122261 TI - Egalitarian and the case of the missing link. PMID- 15122262 TI - Human disease: the centrosome connection. PMID- 15122263 TI - New candidates for vesicle coat proteins. PMID- 15122264 TI - Fatal liaisons of p53 with Bax and Bak. PMID- 15122265 TI - A Bit-role for integrins in apoptosis. PMID- 15122266 TI - Actin dynamics: re-drawing the map. PMID- 15122268 TI - In vivo self-renewing divisions of haematopoietic stem cells are increased in the absence of the early G1-phase inhibitor, p18INK4C. AB - Self-renewal of stem cells is critical for tissue repair and maintenance of organ integrity in most mammalian systems. The relative asymmetry between self-renewal and differentiation in balance with apoptosis determines the size and durability of a stem-cell pool. Regulation of the cell cycle is one of the fundamental mechanisms underlying determination of cell fate. Absence of p21(Cip1/Waf1), a late G1-phase cyclin-dependent kinase inhibitor (CKI), has previously been shown to enable cell-cycle entry of haematopoietic stem cells, but leads to premature exhaustion of the stem cells under conditions of stress. We show here that deletion of an early G1-phase CKI, p18(INK4C), results in strikingly improved long-term engraftment, largely by increasing self-renewing divisions of the primitive cells in murine transplant models. Therefore, different CKIs have highly distinct effects on the kinetics of stem cells, possibly because of their active position in the cell cycle, and p18(INK4C) appears to be a strong inhibitor limiting the potential of stem-cell self-renewal in vivo. PMID- 15122269 TI - An epididymis-specific beta-defensin is important for the initiation of sperm maturation. AB - Although the role of the epididymis, a male accessory sex organ, in sperm maturation has been established for nearly four decades, the maturation process itself has not been linked to a specific molecule of epididymal origin. Here we show that Bin1b, a rat epididymis-specific beta-defensin with antimicrobial activity, can bind to the sperm head in different regions of the epididymis with varied binding patterns. In addition, Bin1b-expressing cells, either of epididymal origin or from a Bin1b-transfected cell line, can induce progressive sperm motility in immotile immature sperm. This induction of motility is mediated by the Bin1b-induced uptake of Ca(2+), a mechanism that has a less prominent role in maintaining motility in mature sperm. In vivo antisense experiments show that suppressed expression of Bin1b results in reduced binding of Bin1b to caput sperm and in considerable attenuation of sperm motility and progressive movement. Thus, beta-defensin is important for the acquisition of sperm motility and the initiation of sperm maturation. PMID- 15122271 TI - NIH under scrutiny. PMID- 15122272 TI - Cautious exuberance after the first quarter. PMID- 15122273 TI - Bioinformatics attracts big guns. PMID- 15122274 TI - EU enlargement brings biotech opportunities. PMID- 15122275 TI - FDA plans to improve and accelerate product reviews. PMID- 15122276 TI - US NIH draft guidelines threaten diagnostics sector. PMID- 15122278 TI - EU expansion: enlarged horizons or false dawn? PMID- 15122279 TI - Phage offer a real alternative. PMID- 15122281 TI - Drugs in crops. PMID- 15122282 TI - Drugs in crops. PMID- 15122283 TI - Drugs in crops. PMID- 15122284 TI - Science, politics and the President's Council on Bioethics. PMID- 15122285 TI - Putting biotech risk in its place. PMID- 15122286 TI - Structure-aided drug design's next generation. PMID- 15122287 TI - Next-generation protein drugs. PMID- 15122288 TI - Overcoming the gridlock in discovery research. PMID- 15122289 TI - Genomics taken to the extreme. PMID- 15122290 TI - Teamwork in phytoremediation. PMID- 15122291 TI - Putting the rat on the map. PMID- 15122293 TI - Alternative splicing in disease and therapy. AB - Alternative splicing is the major source of proteome diversity in humans and thus is highly relevant to disease and therapy. For example, recent work suggests that the long-sought-after target of the analgesic acetaminophen is a neural-specific, alternatively spliced isoform of cyclooxygenase 1 (COX-1). Several important diseases, such as cystic fibrosis, have been linked with mutations or variations in either cis-acting elements or trans-acting factors that lead to aberrant splicing and abnormal protein production. Correction of erroneous splicing is thus an important goal of molecular therapies. Recent experiments have used modified oligonucleotides to inhibit cryptic exons or to activate exons weakened by mutations, suggesting that these reagents could eventually lead to effective therapies. PMID- 15122294 TI - Retraction. Fluorobodies combine GFP fluorescence with the binding characteristics of antibodies. PMID- 15122295 TI - Effecting a comprehensive intellectual property strategy using the Madrid Protocol. PMID- 15122296 TI - Recent patent applications related to molecular cloning. PMID- 15122299 TI - Data standards for 'omic' science. PMID- 15122300 TI - Microarray reality checks in the context of a complex disease. AB - A problem in analyzing microarray-based gene expression data is the separation of genes causally involved in a disease from innocent bystander genes, whose expression levels have been secondarily altered by primary changes elsewhere. To investigate this issue systematically in the context of a class of complex human diseases, we have compared microarray-based gene expression data with non microarray-based clinical and biological data about the schizophrenias to ask whether these two approaches prioritize the same genes. We find that genes whose expression changes are deemed to be of importance from microarrays are rarely those classified as of importance from clinical, in situ, molecular, single nucleotide polymorphism (SNP) association, knockout and drug perturbation data. This disparity is not limited to the schizophrenias but characterizes other human disease data sets. It also extends to biological validation of microarray data in model organisms, in which genome-wide phenotypic data have been systematically compared with microarray data. In addition, different bioinformatic protocols applied to the same microarray data yield quite different gene sets and thus make clinical decisions less straightforward. We discuss how progress may be improved in the clinical area by the assignment of high-quality phenotypic values to each member of a microarray-assigned gene set. PMID- 15122301 TI - International benchmarking of biotech research centers--lessons and perspectives. PMID- 15122302 TI - Genome sequence of the lignocellulose degrading fungus Phanerochaete chrysosporium strain RP78. AB - White rot fungi efficiently degrade lignin, a complex aromatic polymer in wood that is among the most abundant natural materials on earth. These fungi use extracellular oxidative enzymes that are also able to transform related aromatic compounds found in explosive contaminants, pesticides and toxic waste. We have sequenced the 30-million base-pair genome of Phanerochaete chrysosporium strain RP78 using a whole genome shotgun approach. The P. chrysosporium genome reveals an impressive array of genes encoding secreted oxidases, peroxidases and hydrolytic enzymes that cooperate in wood decay. Analysis of the genome data will enhance our understanding of lignocellulose degradation, a pivotal process in the global carbon cycle, and provide a framework for further development of bioprocesses for biomass utilization, organopollutant degradation and fiber bleaching. This genome provides a high quality draft sequence of a basidiomycete, a major fungal phylum that includes important plant and animal pathogens. PMID- 15122303 TI - Overexpression of lipoprotein lipase in transgenic rabbits leads to increased small dense LDL in plasma and promotes atherosclerosis. AB - Lipoprotein lipase (LPL) is a key enzyme in the hydrolysis of triglyceride-rich lipoproteins. Previous studies using transgenic mice and rabbits have demonstrated that high level of LPL activity in adipose and skeletal muscle protects against diet-induced hypercholesterolemia and subsequently prevents aortic atherosclerosis. However, it is unknown, per se, whether increased LPL activity itself is antiatherogenic, or whether the antiatherogenic effect of LPL is dependent upon the LPL lipid-lowering effect. To address this issue, we fed LPL transgenic and littermate rabbits diets containing different amounts of cholesterol (0.3-0.6%) adjusted to maintain their plasma cholesterol concentrations at similarly high levels for 16 weeks. We analyzed their lipoprotein profiles and compared their susceptibility to atherosclerosis. The results showed that the overexpression of LPL in transgenic rabbits reduced remnant lipoproteins (beta-VLDL, d<1.006 g/ml) but concomitantly led to a significant increase of the large (d=1.02-1.04 g/ml) and small LDLs (d=1.04-1.06 g/ml) compared to the amounts in control rabbits. Furthermore, we found that with equally high hypercholesterolemia, transgenic rabbits developed 1.8-fold more extensive aortic atherosclerosis than control rabbits. To examine the hypothesis that altered lipoprotein profiles may be responsible for the enhanced atherosclerosis in transgenic rabbits, we studied the atherogenic properties of apoB-containing lipoproteins in vitro. These studies revealed that small-sized LDLs of transgenic rabbits were more susceptible to copper-induced oxidation and had higher affinity to biglycan than large remnant lipoproteins. We conclude, therefore, that LPL exerts a dual function in terms of its atherogenicity, namely antiatherogenicity, through enhancing receptor-mediated remnant lipoprotein catabolism and proatherogenicity via the generation of a large amount of small sized LDLs. At an equal atherogenic-cholesterol level, small and dense LDLs are more atherogenic than large remnant lipoproteins. PMID- 15122304 TI - Deposition and passage of transthyretin through the blood-nerve barrier in recipients of familial amyloid polyneuropathy livers. AB - Familial amyloid polyneuropathy (FAP) is characterized by deposition of mutated transthyretin (TTR) in the peripheral nervous system. Prior to amyloid fibrils, nonfibrillar TTR aggregates are deposited inducing oxidative stress with increased nitration (3-NT). As the major source of TTR is the liver, liver transplantation (LT) is used to halt FAP. Given the shortage of liver donors, domino LT (DLT) using FAP livers is performed. The correlation between TTR deposition in the skin and nerve was tested in biopsies from normal individuals, asymptomatic carriers (FAP 0) and FAP patients; in FAP 0, nonfibrillar TTR was observed both in the skin and nerve in the same individuals; in patients, amyloid was detected in both tissues. The occurrence of amyloidosis in recipients of FAP livers was evaluated 1-7 years after DLT: TTR deposition occurred in the skin 3 years after transplantation either as amyloid or aggregates; in one of the recipients, fibrillar TTR was present in the epineurium 6 years after DLT. Deposits were scarce and 3-NT immunostaining was irrelevant. Nerve biopsies from DLT recipients had no FAP-related neuropathy. Our findings suggest that TTR amyloid formation occurs faster than predicted and that TTR of liver origin can cross the blood-nerve barrier. Recipients of FAP livers should be under surveillance for TTR deposition and tissue damage. PMID- 15122305 TI - Aberrant CpG island hypermethylation of multiple genes in colorectal neoplasia. AB - CpG island hypermethylation is a potential means of inactivating tumor suppressor genes, and many genes have been demonstrated to be hypermethylated and silenced in colorectal cancer. However, limited data is available upon the concurrent methylation of multiple genes in colorectal cancer and in its precursor lesion. To address changes in the methylation profiles of multiple genes during colorectal carcinogenesis, we investigated the methylation of 12 genes (APC, COX 2, DAP-kinase, E-cadherin, GSTP1, hMLH1, MGMT, p14, p16, RASSF1A, THBS1, and TIMP3) in normal colon (n=24), colon adenoma (n=95), and colorectal cancer (n=149), using methylation-specific PCR. The average number of these genes methylated per sample was 0.12, 1.8, and 3.0 in normal colon mucosa, adenoma, and carcinoma, respectively, showing a stepwise increase (P<0.001). All the genes were methylated in colorectal cancer at frequencies varying from 51 to 9.4% and colon adenoma displayed methylation for the 11 genes, except for GSTP1, at frequencies varying from 40 to 1.1%. In contrast, normal colon mucosa demonstrated methylation for APC only, at a frequency of 12.5%. The total number of methylated genes per tumor showed a continuous, nonbimodal distribution in colon adenoma or cancer. CpG island hypermethylation exhibited a proclivity toward proximal colon cancer or adenoma, and the average number of genes methylated was higher in proximal colon cancer or adenoma than in distal colon cancer or adenoma, respectively (3.5 vs 2.6, P=0.018 for cancer, and 2.5 vs 1.4, P=0.003 for adenoma). In conclusion, concurrent CpG island methylation is an early and frequent event during colorectal carcinogenesis. It appears that CpG island methylation plays a more important role in proximal colon cancer development than in distal colon cancer development. PMID- 15122306 TI - Asbestos-derived reactive oxygen species activate TGF-beta1. AB - Transforming growth factor-beta1 (TGF-beta1) is a potent peptide that inhibits epithelial and mesenchymal cell proliferation and stimulates the synthesis of extracellular matrix components. This cytokine is produced in a biologically latent complex bound to a latent-associated peptide (LAP), and it is the disassociation of this complex that regulates TGF-beta activity. A number of mechanisms have been shown to activate TGF-beta1. We show here that reactive oxygen species (ROS), generated by the iron in chrysotile or crocidolite asbestos, mediate the biological activity of TGF-beta1. Recombinant human latent TGF-beta1 was activated in a cell free system in the presence of asbestos and ascorbic acid. Latent TGF-beta1 was overexpressed in both A549 and mink lung epithelial cell lines through an adenovirus vector containing the full-length construct for porcine TGF-beta1. This latent TGF-beta1 was activated in a concentration-dependant fashion by introducing asbestos into the cell cultures. This activation was reduced significantly through the use of superoxide dismutase, catalase or deferoxamine. Amino-acid constituents of the LAP were oxidized as demonstrated by the appearance of carbonyls detected by Western analysis. The oxidized LAP could no longer form a complex with TGF-beta1. Our data support the postulate that ROS derived from asbestos provide a mechanism for activating TGF-beta1 in the alveolar environment by oxidizing amino acids in LAP. PMID- 15122307 TI - Accumulation of prion protein in muscle fibers of experimental chloroquine myopathy: in vivo model for deposition of prion protein in non-neuronal tissues. AB - Prion protein (PrP) is known to accumulate in some non-neuronal tissues under conditions unrelated to prion diseases. The biochemical and biological nature of such accumulated PrP molecules, however, has not been fully evaluated. In this study, we established experimental myopathy in hamsters by long-term administration of chloroquine, and we examined the nature of the PrP molecules that accumulated. PrP accumulation was immunohistochemically demonstrated in autophagic vacuoles in degenerated muscle fibers, and this was accompanied by the accumulation of other molecules related to the neuropathogenesis of prion diseases such as clathrin, cathepsin B, heparan sulfate, and apolipoprotein J. Accumulated PrP molecules were partially insoluble in detergent solution and were slightly less sensitive to proteinase K digestion than normal cellular PrP. Muscle homogenates containing these PrP molecules did not cause disease in inoculated hamsters. The findings indicate that the PrP molecules that accumulated in muscle fibers have distinct biochemical and biological properties. Therefore, experimental chloroquine myopathy is a novel and useful model to investigate the mechanism of deposition of PrP in non-neuronal tissues and might provide new insights in the pathogenesis of prion diseases. PMID- 15122308 TI - Prospective evaluation of blood concentration of mitochondrial DNA as a marker of toxicity in 157 consecutively recruited untreated or HAART-treated HIV-positive patients. AB - Highly active antiretroviral therapy (HAART) can cause mitochondrial toxicity. The concentration of mitochondrial DNA (mtDNA) in peripheral blood cells has been reported to be a marker of this toxicity. However, these observations are controversial and were drawn from small series. Thus, we analysed the value of blood mtDNA as a marker of mitochondrial toxicity in a large cohort of human immunodeficiency virus (HIV)-infected out-patients during routine clinical evaluations. Real-time quantitative PCR was used to determine the mtDNA to nuclear DNA (nDNA) ratio in peripheral blood mononuclear cells from 157 consecutive HIV-1-infected patients (13 naive, 144 receiving HAART) and 30 HIV-1 uninfected patients. The mtDNA to nDNA ratio was significantly lower in both groups of HIV-infected patients than in the control group. No significant difference was observed between treated and naive HIV-infected patients. Lactataemia was significantly lower in controls than in the group of HIV-treated patients. None of the treated patients had lactataemia >5 mmol/l or bicarbonates <20 mmol/l. Triglyceride levels were significantly higher in the HAART-treated patients than in the nontreated patients. Clinical symptoms of lipodystrophy were observed in 62 HAART-treated patients. These symptoms were not associated with an abnormal mtDNA to nDNA ratio or plasma triglyceride concentration. The mtDNA to nDNA ratio was lower in DDI/D4T-treated patients than in AZT/3TC-treated patients. In conclusion, there are no obvious links between the mtDNA to nDNA ratio in peripheral mononuclear cells and any clinical symptoms or lactate level. Thus, the mtDNA to nDNA ratio in leukocytes does not seem to be an accurate marker of mild and/or long-term mitochondrial toxicity. PMID- 15122309 TI - Abnormal retinal vascular development in IL-18 knockout mice. AB - Recent studies have indicated that interleukin 18 (IL-18) might act as either an angiogenic or an angiostatic factor, but the true function of this protein in vascular development is unclear. We therefore investigated the role of IL-18 in the formation of retinal vessels. Development of the retinal vasculature was compared in IL-18 knockout (KO) and wild-type (WT) mice at several different time points. The formation of vessels was evaluated using angiography of flat-mounted retinal samples after inoculation with fluorescein dextran. Retinal samples from both groups were also evaluated through histological examinations, and the expression of angiogenic factors was examined using the reverse-transcription polymerase chain reaction. The capillary retinal vessels in both WT and IL-18 KO mice had reached the peripheral retina by postnatal day (P) 7. However, IL-18 KO mice showed angiectasis and vascular leakage at P7, especially in the mid peripheral retina. These symptoms were not observed in WT mice at any stage. Histopathological analysis confirmed abnormal vascular formation in IL-18 KO mice at P14. Interestingly, these abnormalities regressed over time and had disappeared by P84. Several angiogenesis-associated factors, including vascular endothelial growth factor (VEGF), basic fibroblast-growth factor (bFGF), platelet derived growth factor (PDGF) and pigment epithelium-derived factor (PEDF), were overexpressed in the retinas of IL-18 KO mice compared with those of WT mice at P14. Interferon-gamma was detected only in WT mouse retinas at P14. These results provide new evidence for the role of IL-18 in retinal vascular development. PMID- 15122310 TI - Comparison of 100-day mortality rates associated with i.v. busulfan and cyclophosphamide vs other preparative regimens in allogeneic bone marrow transplantation for chronic myelogenous leukemia: Bayesian sensitivity analyses of confounded treatment and center effects. AB - We evaluated the 100-day mortality rates associated with busulfan-based myeloablative conditioning regimens based on data from 1812 chronic myelogenous leukemia patients who underwent allogeneic blood or marrow transplantation (allotx). In all, 47 patients received intravenous (i.v.) busulfan and cyclophosphamide (i.v.BuCy2) with allotx at MD Anderson Cancer Center (MDACC) during 1995-1999. The remaining 1765 patients, whose data were supplied by the International Bone Marrow Transplant Registry (IBMTR), received alternative preparative regimens, primarily Cy-total body irradiation ( approximately 45%) or oral BuCy ( approximately 35%) during 1997-1998. As patients were not randomized between conditioning regimens, the i.v.BuCy2-versus-alternative treatment effect is confounded with a possible center effect due to nontreatment differences associated with factors differing between MDACC and the IBMTR centers. Additional complications are that the i.v.BuCy2-MDACC patients all survived 100 days, and three prognostic subgroups were included. Bayesian sensitivity analyses were performed to assess treatment effect on the probability of 100-day mortality, over a range of possible MDACC-versus-IBMTR center effects. For these patients, the posterior probability that i.v.BuCy2 was superior to alternative conditioning regimens ranges from 0.54 to 0.99, depending on prognosis and the magnitude of the assumed center effect. PMID- 15122311 TI - Analysis of 96 patients with advanced ovarian carcinoma treated with high-dose chemotherapy and autologous stem cell transplantation. AB - The purpose of this study was to identify characteristics significant to survival and progression-free survival in patients with advanced ovarian cancer receiving high-dose chemotherapy. In all, 96 patients received autologous stem cell transplantation. Regimens included paclitaxel with carboplatin (PC), topotecan, melphalan, cyclophosphamide (TMC) and cyclophosphamide, BCNU, thiotepa (CBT). At the time of transplantation, 43% of patients were in clinical CR, 34% were in clinical PR, 18% had progressive disease and 5% had stable disease. There were no treatment-related deaths. The 6-year survival by Kaplan-Meier was 38%. For patients who received transplantation for remission consolidation, the 6-year survival was 53% with a PFS of 29%. On univariate analysis, the CBT regimen, clear cell histology and disease status other than CR prior to treatment were statistically significant adverse prognostic factors. This analysis has demonstrated that patients in clinical remission are most likely to benefit from autologous transplantation, with the exception of patients with clear cell histology. The TMC combination appeared to be superior to the PC and CBT combinations. Comparative studies of different consolidation approaches will be necessary to determine if autologous transplantation is the preferred treatment for this patient population. PMID- 15122312 TI - Safety of the concomitant use of caspofungin and cyclosporin A in patients with invasive fungal infections. AB - Caspofungin, an echinocandin antifungal agent, is active against invasive Aspergillus and Candida infections. In a phase I study in healthy volunteers, mild transient increases in serum aminotransferases were observed with the concomitant administration of caspofungin and cyclosporin A (CsA). As a result, it is recommended that the concomitant use of the two drugs be limited to those settings with appropriate risk-benefit balance. We retrospectively assessed safety data in 14 patients with refractory invasive mycoses who were treated concomitantly with CsA and caspofungin before the drug was licensed in Spain. In all, 13 patients were adults (median age, 31.5 years; range, 14-67 years). The average duration of concomitant therapy was 15 days (range, 2-43 days). No clinically significant elevations of serum aminotransferases were observed, and no patient had concomitant therapy discontinued or interrupted due to a drug related adverse event. In this study of a limited number of patients, the coadministration of caspofungin and CsA was generally well tolerated. PMID- 15122313 TI - Mcl-1 is required for Akata6 B-lymphoma cell survival and is converted to a cell death molecule by efficient caspase-mediated cleavage. AB - Enforced expression of the antiapoptotic Bcl-2 family protein Mcl-1 promotes lymphomagenesis in the mouse; however, the functional role of Mcl-1 in human B cell lymphoma remains unclear. We demonstrate that Mcl-1 is widely expressed in malignant B-cells, and high-level expression of Mcl-1 is required for B-lymphoma cell survival, since transfection of Mcl-1-specific antisense oligodeoxynucleotides was sufficient to promote apoptosis in Akata6 lymphoma cells. Mcl-1 was efficiently cleaved by caspases at evolutionarily conserved aspartic acid residues in vitro, and during cisplatin-induced apoptosis in B lymphoma cell lines and spontaneous apoptosis of primary malignant B-cells. Overexpression of the Mcl-1 cleavage product that accumulated during apoptosis was sufficient to kill cells. Therefore, Mcl-1 is an essential survival molecule for B-lymphoma cells and is cleaved by caspases to a death-promoting molecule during apoptosis. In contrast to Mcl-1, Bcl-2 and Bcl-XL were relatively resistant to caspase cleavage in vitro and in intact cells. Interfering with Mcl 1 function appears to be an effective means of inducing apoptosis in Mcl-1 positive B-cell lymphoma, and the unique sensitivity of Mcl-1 to caspase-mediated cleavage suggests an attractive strategy for converting it to a proapoptotic molecule. PMID- 15122314 TI - Alternative splice variant of actinin-4 in small cell lung cancer. AB - Tumor-associated alternative RNA splicing has gained considerable attention. We identified a novel alternative splice variant RNA of actinin-4 in human small cell lung cancer (SCLC). Expression of the splice variant was highly specific to SCLC cell lines (10/10), biopsies (3/3), and testis. The variant encoded a peptide with a three amino-acid change in exon 8, where the germline missense mutation takes place in familial focal segmental glomerulosclerosis (FSGS). The variant protein showed high affinity to filamentous actin polymers and was not localized with cortical actin. Alternatively spliced actinin-4 may be a new diagnostic marker of SCLC and a candidate target for selective therapy. PMID- 15122315 TI - HIPK2 neutralizes MDM2 inhibition rescuing p53 transcriptional activity and apoptotic function. AB - The p53 oncosuppressor protein is subject to negative regulation by MDM2, which efficiently inhibits its activity through an autoregulatory loop. In response to stress, however, p53 undergoes post-translational modifications that allow the protein to escape MDM2 control, accumulate, and become active. Recent studies have shown that, following DNA damage, the HIPK2 serine/threonine kinase binds and phosphorylates p53, inducing p53 transcriptional activity and apoptotic function. Here, we investigated the role of HIPK2 in the activation of p53 in the presence of MDM2. We found that HIPK2 rescues p53 transcriptional activity overcoming MDM2 inhibition, and that restoration of this p53 function induces apoptosis. Recovery of p53-dependent apoptosis is achieved by preventing p53 nuclear export and ubiquitination mediated by MDM2 in vitro and in vivo following genotoxic stress. These results shed new light on the mechanisms by which the HIPK2/p53 pathway promotes apoptosis and suppression of tumorigenesis. PMID- 15122317 TI - Inhibition of in vivo breast cancer growth by antisense oligodeoxynucleotides to type I insulin-like growth factor receptor mRNA involves inactivation of ErbBs, PI-3K/Akt and p42/p44 MAPK signaling pathways but not modulation of progesterone receptor activity. AB - The present study addresses the effect of targeting type I insulin-like growth factor receptor (IGF-IR) with antisense strategies in in vivo growth of breast cancer cells. Our research was carried out on C4HD tumors from an experimental model of hormonal carcinogenesis in which the synthetic progestin medroxyprogesterone acetate (MPA) induced mammary adenocarcinomas in Balb/c mice. We employed two different experimental strategies. With the first one we demonstrated that direct intratumor injection of phosphorothioate antisense oligodeoxynucleotides (AS[S]ODNs) to IGF-IR mRNA resulted in a significant inhibition of C4HD tumor growth. In the second experimental strategy, we assessed the effect of intravenous (i.v.) injection of AS [S]ODN on C4HD tumor growth. This systemic treatment also resulted in significant reduction in tumor growth. The antitumor effect of IGF-IR AS[S]ODNs in both experimental protocols was due to a specific antisense mechanism, since growth inhibition was dose-dependent and no abrogation of tumor proliferation was observed in mice treated with phosphorothioate sense ODNs (S[S]ODNs). In addition, IGF-IR expression was inhibited in tumors from mice receiving AS[S]ODNs, as compared to tumors from control groups. We then investigated signal transduction pathways modulated in vivo by AS[S]ODNs treatment. Tumors from AS[S]ODN-treated mice of both intratumoral and intravenous protocols showed a significant decrease in the degree of insulin receptor substrate-1 (IRS-1) tyrosine phosphorylation. Activation of two of the main IGF-IR signaling pathways, phosphatidylinositol 3 kinase (PI-3K)/Akt and p42/p44 mitogen-activated protein kinases (MAPK) was abolished in tumors growing in AS[S]ODN-treated animals. Moreover, ErbB-2 tyrosine phosphorylation was blocked by in vivo administration of AS[S]ODNs. On the other hand, we found no regulation of either progesterone receptor expression or activity by in vivo AS[S]ODNs administration. Our results for the first time demonstrated that breast cancer growth can be inhibited by direct in vivo administration of IGF-IR AS[S]ODNs. PMID- 15122316 TI - Human hRad1 but not hRad9 protects hHus1 from ubiquitin-proteasomal degradation. AB - Three of the Rad family proteins, Rad9, Rad1, and Hus1, can interact with each other and form a heterotrimeric complex that is thought to play a role in the sensing step of the DNA integrity checkpoint pathways, but the nature of the Rad9 Rad1-Hus1 complex assembly remains enigmatic. Here, we demonstrate that the human hRad1 protein plays a significant role as molecular chaperone in the process of the hRad9-hRad1-hHus1 heterotrimeric complex formation. In contrast to hRad1, hHus1 is an unstable protein that is actively degraded via the ubiquitin proteasome pathway. We show that treating cells with proteasome-specific inhibitors stabilizes hHus1 expression. Moreover, hRad1 can associate with hHus1 in the absence of hRad9 and protect hHus1 from ubiquitination and degradation in the cytoplasm. Importantly, genotoxic stress induces hRad1 expression and stabilizes the hHus1 protein. Taken together, these findings suggest a novel role of hRad1 as a potential intrinsic chaperone in the stabilization of hHus1 for the hRad9-hRad1-hHus1 checkpoint complex formation. PMID- 15122318 TI - Myeloid Leukemia Factor 1 inhibits erythropoietin-induced differentiation, cell cycle exit and p27Kip1 accumulation. AB - Myeloid leukemia factor 1 (MLF1) is a novel oncoprotein involved in translocations associated with acute myeloid leukemia (AML), especially erythroleukemias. In this study, we demonstrate that ectopic expression of Mlf1 prevented J2E erythroleukemic cells from undergoing biological and morphological maturation in response to erythropoietin (Epo). We show that Mlf1 inhibited Epo induced cell cycle exit and suppressed a rise in the cell cycle inhibitor p27(Kip1). Unlike differentiating J2E cells, Mlf1-expressing cells did not downregulate Cul1 and Skp2, components of the ubiquitin E3 ligase complex SCF(Skp2) involved in the proteasomal degradation of p27(Kip1). In contrast, Mlf1 did not interfere with increases in p27(Kip1) and terminal differentiation initiated by thyroid hormone withdrawal from erythroid cells, or cytokine stimulated maturation of myeloid cells. These data demonstrate that Mlf1 interferes with an Epo-responsive pathway involving p27(Kip1) accumulation, which inhibits cell cycle arrest essential for erythroid terminal differentiation. PMID- 15122320 TI - Make WARTS, not cancer! AB - The WARTS gene encodes a kinase that localizes to the mitotic apparatus of a dividing cell. Named WARTS after the growths that develop in the eyes of Drosophila in which the gene is deleted. WARTS is also implicated as a tumor suppressor in mice and humans. In this issue of Oncogene, Iida et al. describe experiments suggesting that, in addition to a role in regulating mitosis, WARTS functions to prevent further rounds of DNA synthesis and mitosis in tetraploid cells. As well as opening up new possibilities of exploring the as yet ill defined mechanistic basis of the tetraploidy checkpoint, the involvement of a tumor-suppressor gene in this checkpoint supports its importance as a safeguard against the acquisition of genomic instability, a key event in the progression to cancer. PMID- 15122319 TI - Modulations of critical cell cycle regulatory events during chemoprevention of ultraviolet B-mediated responses by resveratrol in SKH-1 hairless mouse skin. AB - Multiple exposures to solar ultraviolet (UV) radiation cause critical damages that may lead to the development of several cutaneous disorders including skin cancer, the most frequently diagnosed malignancy in the USA. Therefore, efforts are needed to: (i) study the mechanism(s) of UV-mediated cutaneous damages, and (ii) design novel approaches for the management of skin cancer. 'Chemoprevention' via plant-based agents may be a useful approach for the management of neoplasia. Here, we evaluated the involvement of cell cycle regulatory molecules during resveratrol-mediated protection from multiple exposures of UVB (180 mJ/cm(2); on alternate days x 7 exposures) radiations in the SKH-1 hairless mouse skin. Resveratrol was topically applied on the skin of SKH-1 hairless mice at a dose of 10 micromol/mouse (in 0.2 ml acetone; 30 min prior to each UVB exposure). Studies were performed at 24 h following the last UVB exposure. Topical application of resveratrol resulted in significant decrease in UVB-induced bi-fold skin thickness, hyperplasia, and infiltration of leukocytes. The data from immunoblot and/or immunohistochemical analyses revealed that multiple exposure to UVB radiations causes significant upregulation in: (i) proliferating cell nuclear antigen (PCNA), a marker of cellular proliferation, and (ii) cyclin-dependent kinase (cdk)-2, -4 and -6, cyclin-D1, and cyclin-D2. Resveratrol treatment resulted in significant downregulation in UV-mediated increases in these critical cell cycle regulatory proteins. An interesting observation of this study was that resveratrol treatment resulted in a further stimulation of UVB-mediated increases in cyclin kinase inhibitor WAF1/p21 and tumor suppressor p53. Further, resveratrol was also found to cause significant decreases in UVB-mediated upregulation of: (i) the mitogen-activated protein kinase kinase, and (ii) the 42 kDa isotype of mitogen-activated protein kinase (MAPK). Thus, our data suggested that the antiproliferative effects of resveratrol might be mediated via modulation in the expression and function of cell cycle regulatory proteins cyclin-D1 and -D2, cdk-2, -4 and -6, and WAF1/p21. Our data further suggest that the modulation of cki-cyclin-cdk network by resveratrol may be associated with inhibition of the MAPK pathway. We suggest that resveratrol may be useful for the prevention of UVB-mediated cutaneous damages including skin cancer. PMID- 15122321 TI - A novel crosstalk mechanism between nuclear receptor-mediated and growth factor/Ras-mediated pathways through PNRC-Grb2 interaction. AB - It has been demonstrated that proline-rich nuclear receptor coregulatory protein (PNRC) is a nuclear receptor coactivator that interacts with nuclear receptors through an SH3-binding motif located in its C-terminus. In the present report, a physical interaction between PNRC and Grb2 (an adapter protein involved in growth factor/Ras-mediated pathways) has been demonstrated using the GST pull-down assay, the yeast two-hybrid assay, as well as by coimmunoprecipitation. Cotransfection and fluorescence imaging have also confirmed the colocalization of PNRC and Grb2 in mammalian cells. Transient transfection experiments have demonstrated that, by interacting with each other, Grb2 decreases the coactivator activity of PNRC for nuclear receptors, and that PNRC suppresses Grb2-mediated Ras/MAP-kinase activation. Furthermore, it was discovered that HeLa cells overexpressing PNRC grew more slowly when compared to matched controls. Additionally, using a RT-PCR analysis of mRNA on six pairs of cancer/noncancer tissues, PNRC expression was found to be significantly lower in breast cancer tissue than in noncancer tissue. Based on these findings, we believe that PNRC and Grb2, by interacting with each other, can suppress nuclear receptor-mediated regulation and growth factor-mediated regulation in human breast tissue. This is a newly identified crosstalk mechanism for modulating these two important types of regulatory pathways. PMID- 15122322 TI - Extracellular S100A4(mts1) stimulates invasive growth of mouse endothelial cells and modulates MMP-13 matrix metalloproteinase activity. AB - S100A4(mts1) protein expression has been strongly associated with metastatic tumor progression. It has been suggested as a prognostic marker for a number of human cancers. It is proposed that extracellular S100A4 accelerates cancer progression by stimulating the motility of endothelial cells, thereby promoting angiogenesis. Here we show that in 3D culture mouse endothelial cells (SVEC 4-10) respond to recombinant S100A4 by stimulating invasive growth of capillary-like structures. The outgrowth is not dependent on the stimulation of cell proliferation, but rather correlates with the transcriptional modulation of genes involved in the proteolytic degradation of extracellular matrix (ECM). Treatment of SVEC 4-10 with the S100A4 protein leads to the transcriptional activation of collagenase 3 (MMP-13) mRNA followed by subsequent release of the protein from the cells. Beta-casein zymography demonstrates enhancement of proteolytic activity associated with MMP-13. This observation indicates that extracellular S100A4 stimulates the production of ECM degrading enzymes from endothelial cells, thereby stimulating the remodeling of ECM. This could explain the angiogenic and metastasis-stimulating activity of S100A4(mts1). PMID- 15122323 TI - Mitochondrial pro-apoptotic ARTS protein is lost in the majority of acute lymphoblastic leukemia patients. AB - Acquired resistance towards apoptosis is the hallmark of most if not all types of cancer. We have previously identified and characterized ARTS, a broadly expressed protein localized to mitochondria. ARTS was initially shown to mediate TGF-beta induced apoptosis. Recently, we have found that high levels of ARTS induce apoptosis without additional pro-apoptotic stimuli. Further, ARTS promotes apoptosis in response to a wide variety of pro-apoptotic stimuli. Here, we report that the expression of ARTS is lost in all lymphoblasts of more than 70% of childhood acute lymphoblastic leukemia (ALL) patients. The loss of ARTS is specific, as the related non-apoptotic protein H5, bearing 83% identity to ARTS, is unaffected. During remission, ARTS expression is detected again in almost all patients. Two leukemic cell lines, ALL-1 and HL-60 lacking ARTS, were resistant to apoptotic induction by ara-C. Transfection of ARTS into these cells restored their ability to undergo apoptosis in response to this chemotherapeutic agent. We found that methylation process contributes to the loss of ARTS expression. We conclude that the loss of ARTS may provide a selective advantage for cells to escape apoptosis thereby contributing to their transformation to malignant lymphoblasts. We therefore propose that ARTS can function as a tumor suppressor protein in childhood ALL. PMID- 15122324 TI - Control of cell cycle-dependent degradation of c-Ski proto-oncoprotein by Cdc34. AB - It is known that excess amounts of Ski, or any member of its proto-oncoprotein family, causes disruption of the transforming growth factor beta signal transduction pathway, thus causing oncogenic transformation of cells. Previous studies indicate that Ski is a relatively unstable protein whose expression levels can be regulated by ubiquitin-mediated proteolysis. Here, we investigate the mechanism by which the stability of Ski is regulated. We show that the steady state levels of Ski protein are controlled post-translationally by cell cycle dependent proteolysis, wherein Ski is degraded during the interphase of the cell cycle but is relatively stable during mitosis. Furthermore, we demonstrate that the ubiquitin-conjugating enzyme Cdc34 mediates cell cycle-dependent Ski degradation both in vitro and in vivo. Overexpression of dominant-negative Cdc34 stabilizes Ski and enhances its ability to antagonize TGF-beta signaling. Our data suggest that regulated proteolysis of Ski is one of the key mechanisms that control the threshold levels of this proto-oncoprotein, and thus prevents epithelial cells from becoming TGF-beta resistant. PMID- 15122325 TI - Expression of an amino-terminal BRCA1 deletion mutant causes a dominant growth inhibition in MCF10A cells. AB - Expression of deletion mutants of the breast and ovarian cancer-specific tumor suppressor protein, BRCA1, in the mammary epithelial cell line MCF10A revealed a powerful growth suppressive effect by a mutant that has the amino-terminal 302 amino acids deleted (DeltaN-BRCA1). The growth suppression is associated with an increase in apoptosis and amplification in centrosome number. The growth inhibitory effect of DeltaN-BRCA1 was not observed in cervical epithelial HeLa cells, suggesting that the phenotypes of BRCA1 mutant proteins differ depending on the cell line being tested. An internal domain, including BRCA1 residues 303 1292, caused the suppression of MCF10A cell growth, and the amino terminus of BRCA1 autoinhibited the growth suppression. Single point mutations that disrupted the amino-terminal RING domain of BRCA1 caused significant suppression of growth in MCF10A cells. These results suggest that the proper function of the RING domain, likely to be ubiquitin ligase function, is important in regulating the growth of the mammary epithelial cell line and in autoregulating the powerful internal growth-inhibiting domain of the BRCA1 tumor suppressor. PMID- 15122326 TI - E2F3 amplification and overexpression is associated with invasive tumor growth and rapid tumor cell proliferation in urinary bladder cancer. AB - E2F3 is located in the 6p22 bladder amplicon and encodes a transcription factor important for cell cycle regulation and DNA replication. To further investigate the role of E2F3 in bladder cancer, a tissue microarray containing samples from 2317 bladder tumors was used for gene copy number and expression analysis by means of fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC). E2F3 amplification was strongly associated with invasive tumor phenotype and high tumor grade (P < 0.0001 each). None of 272 pTaG1/G2 tumors, but 35 of 311 pT1-4 carcinomas (11.3%), had E2F3 amplification. A high E2F3 expression level was associated with high grade, advanced stage, and E2F3 gene amplification (P < 0.0001 each). To evaluate whether E2F3 expression correlates with tumor proliferation, the Ki67 labeling index (LI) was analysed for each tumor. There was a strong association between a high Ki67 LI and E2F3 expression (P < 0.0001), which was independent of grade and stage. We conclude that E2F3 is frequently amplified and overexpressed in invasively growing bladder cancer (stage pT1-4). E2F3 expression appears to provide a growth advantage to tumor cells by activating cell proliferation in a subset of bladder tumors. PMID- 15122327 TI - Rho family GTPases cooperate with p53 deletion to promote primary mouse embryonic fibroblast cell invasion. AB - The Rho family GTPases Rac1, RhoA and Cdc42 function as molecular switches that transduce intracellular signals regulating multiple cell functions including gene expression, adhesion, migration and invasion. p53 and its regulator p19Arf, on the other hand, are tumor suppressors that are critical in regulating cell cycle progression and apoptosis. Previously, we have demonstrated that the Rho proteins contribute to the cell proliferation, gene transcription and migration phenotypes unleashed by p19Arf or p53 deletion in primary mouse embryo fibroblasts (MEFs). To further investigate their functional interaction in the present study, we have examined the involvement of Rho signaling pathways in p53-mediated cell invasion. We found that in primary MEFs (1) p53 or p19Arf deficiency led to a marked increase in the number of focal adhesion plaques and fibronectin production, and RhoA, Rac1 and Cdc42 contribute to the p53- and p19Arf-mediated focal adhesion regulation, but not fibronectin synthesis; (2) although endogenous Rac1 activity was required for the p19Arf or p53 deficiency-induced migration phenotype, hyperactive Rho GTPases could not further enhance cell migration, rather they suppressed cell-cell adhesion of p53-/- MEFs; (3) expression of the active mutant of RhoA, Rac1 or Cdc42, but not Ras, promoted an invasion phenotype of p53-/-, not p19Arf-/-, cells; (4) although ROCK activation can partially recapitulate Rho induced invasion phenotype, multiple pathways regulated by RhoA, in addition to ROCK, are required to fully cooperate with p53 deficiency to promote cell invasion; and (5) extracellular proteases produced by the active RhoA-transduced cells are also required for the invasion phenotype of p53-/- cells. Combined with our previous observations, these results strongly suggest that mitogenic activation of Rho family GTPases can cooperate with p53 deficiency to promote primary cell invasion as well as transformation and that multiple signaling components regulated by the Rho proteins are involved in these processes. PMID- 15122328 TI - Overexpression of sprouty 2 inhibits HGF/SF-mediated cell growth, invasion, migration, and cytokinesis. AB - A strict regulation of hepatocyte growth factor/scatter factor (HGF/SF)-Met signaling is essential for its appropriate function. Several negative regulators of Met signaling have been identified. Here we report that human Spry2 is induced by HGF/SF and negatively regulates HGF/SF-Met signaling. We show that overexpression of Spry2 inhibits cell proliferation, anchorage-independent cell growth, and migration in wound-healing and in vitro invasion assays. Measured in an electric cell-substrate impedance sensing biosensor, cell movement is restricted, because Spry2 dramatically facilitates cell attachment and spreading by enhancing focal adhesions and increasing stress fibers. An analysis of cell cycle distribution shows, unexpectedly, that Spry2-GFP cells are polyploid. Thus, as with FGF and EGF receptors, Spry2-GFP tempers downstream Met signaling in addition to its pronounced effect on cell adhesion, and it has properties suitable to be considered a tumor-suppressor protein. PMID- 15122329 TI - Connexin43 pseudogene is expressed in tumor cells and inhibits growth. AB - Pseudogenes are classically thought of as nonfunctional DNA sequences due to their inability to be translated, or to produce a functional protein. Gap junctions, a multiprotein complex made of proteins called connexins, are involved in intercellular communication and are deregulated in many cancers. Connexin43 (Cx43) is the only connexin for which a pseudogene has been reported so far. The Cx43 pseudogene (PsiCx43) has all of the features of an expressed gene. We identified the presence of a PsiCx43 mRNA transcript in several cancer cell lines and in none of the normal mammary epithelial cells studied. Using an in vitro translation assay, we found that the PsiCx43 coding plasmid could be translated into a 43 kDa protein. This was further confirmed by expressing a PsiCx43-green fluorescence protein fusion protein in breast cancer MCF-7 cells. We then examined the functional significance of the PsiCx43. In both MTT growth and colony formation assays, significant growth inhibition was observed, a feature common to cells overexpressing the Cx43 gene. However, using a scrape-loading assay, we could not detect any effect on gap junctional intercellular communication. Based on our findings, PsiCx43 joins and enlarges the thus far restricted group of functionally transcribed and translated pseudogenes. PMID- 15122330 TI - Antitumor activity of IFIX, a novel interferon-inducible HIN-200 gene, in breast cancer. AB - We identified IFIX as a new member of the hematopoietic interferon (IFN) inducible nuclear protein with the 200-amino-acid repeat (HIN-200) family. Six different alternatively spliced forms of mRNA are transcribed from the IFIX gene, which are predicted to encode six different isoforms of IFIX proteins (IFIXalpha1, alpha2, beta1, beta2, gamma1, and gamma2). The IFIX proteins are primarily localized in the nucleus. They share a common N-terminal region that contains a predicted pyrin domain and a putative nuclear localization signal. Unlike IFIXalpha and IFIXbeta, IFIXgamma isoforms do not have the 200-amino-acid signature motif. Interestingly, the expression of IFIX was reduced in most human breast tumors and breast cancer cell lines. Expression of IFIXalpha1, the longest isoform of IFIX, in human breast cancer cell lines reduced their anchorage dependent and -independent growth in vitro and tumorigenicity in nude mice. Moreover, a liposome-mediated IFIXalpha1 gene transfer suppressed the growth of already-formed tumors in a breast cancer xenograft model. IFIXalpha1 appears to suppress the growth of breast cancer cells in a pRB- and p53-independent manner by increasing the expression of the cyclin-dependent kinase inhibitor p21(CIP1), which leads to the reduction of the kinase activity of both Cdk2 and p34(Cdc2). Together, our results show that IFIXalpha1 possesses a tumor-suppressor activity and suggest IFIXalpha1 may be used as a therapeutic agent in cancer treatment. PMID- 15122331 TI - Loss of Rad52 partially rescues tumorigenesis and T-cell maturation in Atm deficient mice. AB - Ataxia Telangiectasia (A-T) is an autosomal recessive disease caused by loss of function of the protein kinase ATM. Atm-deficient mice display several phenotypes consistent with the human disease, including predisposition to cancer, growth retardation, cell-proliferation defects and infertility. A-T patients have a several hundred fold increased risk of developing lymphomas and leukemias, which are typically highly invasive. By reducing homologous recombination through genetic deletion of the Rad52 protein, we were able to decrease substantially the development of T-cell lymphomas in Atm-/- mice, resulting in an increased life span of the double mutant mice. Additionally, we were able to partially rescue the T-cell development of Atm-/- mice. Other phenotypes, including growth defects, genomic instability, infertility and radiosensitivity, were not rescued. Our results suggest that excessive recombination is an important contributor to tumorigenesis in A-T. PMID- 15122332 TI - Inhibition of cathepsin B and MMP-9 gene expression in glioblastoma cell line via RNA interference reduces tumor cell invasion, tumor growth and angiogenesis. AB - Extracellular proteases have been shown to cooperatively influence matrix degradation and tumor cell invasion through proteolytic cascades, with individual proteases having distinct roles in tumor growth, invasion, migration and angiogenesis. Matrix metalloproteases (MMP)-9 and cathepsin B have been shown to participate in the processes of tumor growth, vascularization and invasion of gliomas. In the present study, we used a cytomegalovirus promoter-driven DNA template approach to induce hairpin RNA (hpRNA)-triggered RNA interference (RNAi) to block MMP-9 and cathepsin B gene expression with a single construct. Transfection of a plasmid vector-expressing double-stranded RNA (dsRNA) for MMP-9 and cathepsin B significantly inhibited MMP-9 and cathepsin B expression and reduced the invasive behavior of SNB19, glioblastoma cell line in Matrigel and spheroid invasion models. Downregulation of MMP-9 and cathepsin B using RNAi in SNB19 cells reduced cell-cell interaction of human microvascular endothelial cells, resulting in the disruption of capillary network formation in both in vitro and in vivo models. Direct intratumoral injections of plasmid DNA expressing hpRNA for MMP-9 and cathepsin B significantly inhibited established glioma tumor growth and invasion in intracranial tumors in vivo. Further intraperitoneal (i.p.) injections of plasmid DNA expressing hpRNA for MMP-9 and cathepsin B completely regressed pre-established tumors for a long time (4 months) without any indication of these tumor cells. For the first time, these observations demonstrate that the simultaneous RNAi-mediated targeting of MMP-9 and cathepsin B has potential application for the treatment of human gliomas. PMID- 15122333 TI - Low p21Waf1/Cip1 protein level sensitizes testicular germ cell tumor cells to Fas mediated apoptosis. AB - In the present study, we investigated the relation between p21 expression and the sensitivity of testicular germ cell tumor (TGCT) cells to apoptotic stimuli. Despite similar cisplatin-induced wild-type p53 accumulation, the TGCT cell lines Tera and Scha expressed low p21 protein and mRNA levels in comparison to A2780 ovarian cancer cells. Inhibition of the proteasome complex with MG-132 increased p21 protein levels in TGCT cells but much more in A2780 cells, whereas cisplatin had no additional effect on p21 protein levels. Inhibition of caspase-3 activity in TGCT cells with the broad-spectrum caspase inhibitor zVAD-fmk had no effect on p21 levels and also not upon cisplatin treatment. A similar induction of p53 irradiation, in contrast to cisplatin, substantially increased both p21 mRNA and protein expression in Tera cells. Cisplatin-treated Tera cells expressing low p21 protein levels were Fas-sensitive, while irradiation-induced p21, which was mainly localized in the cytosol, rendered irradiated Tera cells resistant to Fas induced apoptosis. Sensitivity of irradiated Tera cells to Fas-induced apoptosis was restored by short interfering RNA-specific suppression of p21 expression. These results strongly indicate that the low p21 protein levels are caused by reduced p21 gene transcription and sensitize cisplatin-treated TGCT cells to the Fas death pathway. PMID- 15122334 TI - Disruption of MKK4 signaling reveals its tumor-suppressor role in embryonic stem cells. AB - The dual Ser/Thr kinase MKK4 and its downstream targets JNK and p38 regulate critical cellular functions during embryogenesis and development. MKK4 has been identified as a putative tumor-suppressor gene in human solid tumors of breast, prostate and pancreas. To clarify the mechanisms underlying the transforming potential of molecular defects targeting MKK4, we have generated totipotent embryonic stem (ES) cells expressing the dominant-negative mutant DN-MKK4(Ala), S257A/T261A. Stably transfected DN-MKK4-ES cells exhibit a transformed fibroblast like morphology, reduced proliferation rate, were no more submitted to cell contact inhibition, were growing in soft agar, and were much more tumorigenic than parental ES cells in athymic nude mice. These phenotypic changes: (i) are consistent with the protection of DN-MKK4-transfected ES cells from spontaneous, cell density-dependent, and stress-induced apoptosis (DAPI staining and poly (ADP ribose) polymerase (PARP) cleavage) and (ii) correlated with alterations in JNK, p38, and Erk-1/-2 MAPK/SAPK signaling. Taken together, our data provide a new mechanism linking the MKK4 signaling pathways to cancer progression and identify MKK4 as a tumor-suppressor gene implicated in several transforming functions. PMID- 15122335 TI - Tumor suppressor WARTS ensures genomic integrity by regulating both mitotic progression and G1 tetraploidy checkpoint function. AB - Defects in chromosomes or mitotic spindles activate the spindle checkpoint, resulting in cell cycle arrest at prometaphase. The prolonged activation of spindle checkpoint generally leads to mitotic exit without segregation after a transient mitotic arrest and the consequent formation of tetraploid G(1) cells. These tetraploid cells are usually blocked to enter the subsequent S phase by the activation of p53/pRb pathway, which is referred to as the G(1) tetraploidy checkpoint. A human homologue of the Drosophila warts tumor suppressor, WARTS, is an evolutionarily conserved serine-threonine kinase and implicated in development of human tumors. We previously showed that WARTS plays a crucial role in controlling mitotic progression by forming a regulatory complex with zyxin, a regulator of actin filament assembly, on mitotic apparatus. However, when WARTS is activated during cell cycle and how the loss of WARTS function leads to tumorigenesis have not been elucidated. Here we show that WARTS is activated during mitosis in mammalian cells, and that overexpression of a kinase-inactive WARTS in Rat1 fibroblasts significantly induced mitotic delay. This delay resulted from prolonged activation of the spindle assembly checkpoint and was frequently followed by mitotic slippage and the development of tetraploidy. The resulting tetraploid cells then abrogated the G(1) tetraploidy checkpoint and entered S phase to achieve a DNA content of 8N. This impairment of G(1) tetraploidy checkpoint was caused as a consequence of failure to induce p53 expression by expressing a kinase-inactive WARTS. WARTS thus plays a critical role in maintenance of ploidy through its actions in both mitotic progression and the G(1) tetraploidy checkpoint. PMID- 15122336 TI - Novel expression of N-cadherin elicits in vitro bladder cell invasion via the Akt signaling pathway. AB - Novel N-cadherin expression has been linked to the invasive phenotype in bladder tumors yet a primary role for N-cadherin in invasion has not been defined in this model. To address this, N-cadherin was stably transfected into E-cadherin expressing bladder carcinoma cells. This resulted in an enhanced invasive capacity in in vitro assays that was blocked by incubation with an N-cadherin function-blocking antibody in a dose-dependent manner. Analysis of the signaling pathway(s) implicated in N-cadherin-mediated invasion in bladder carcinoma cell lines revealed no correlation between MAPK signaling and invasion, in the presence or absence of fibroblast growth factor 2. Also, while MAPK and p38 kinase inhibitors did not alter the invasive behavior of these cells, an increase in the phosphorylation of Akt at serine-473 was detected in N-cadherin transfectants, suggestive of N-cadherin-mediated Akt activation in bladder cell invasion. Incubation of N-cadherin transfectants with either PI3 kinase or Akt inhibitors resulted in a significant decrease in the invasive capacity of these cells. Exposure of cells to PP2, a src family kinase inhibitor, also decreased the invasive potential of N-cadherin transfectants and resulted in reduced phosphorylation of Akt. The involvement of Akt signaling in bladder cell invasion was also supported by the inhibition of bladder cell invasion by cells constitutively expressing an activated Akt kinase, using the PI3 kinase and Akt inhibitors and PP2. These results suggest that activation of PI3/AKT kinase following N-cadherin expression contributes to the increased invasive potential of bladder carcinoma cells. PMID- 15122337 TI - The candidate tumor suppressor gene BLU, located at the commonly deleted region 3p21.3, is an E2F-regulated, stress-responsive gene and inactivated by both epigenetic and genetic mechanisms in nasopharyngeal carcinoma. AB - Loss of heterozygosity at 3p21 is common in various cancers including nasopharyngeal carcinoma (NPC). BLU is one of the candidate tumor suppressor genes (TSGs) in this region. Ectopic expression of BLU results in the inhibition of colony formation of cancer cells, suggesting that BLU is a tumor suppressor. We have identified a functional BLU promoter and found that it can be activated by environmental stresses such as heat shock, and is regulated by E2F. The promoter and first exon are located within a CpG island. BLU is highly expressed in testis and normal upper respiratory tract tissues including nasopharynx. However, in all seven NPC cell lines examined, BLU expression was downregulated and inversely correlated with promoter hypermethylation. Biallelic epigenetic inactivation of BLU was also observed in three cell lines. Hypermethylation was further detected in 19/29 (66%) of primary NPC tumors, but not in normal nasopharyngeal tissues. Treatment of NPC cell lines with 5-aza-2'-deoxycytidine activated BLU expression along with promoter demethylation. Although hypermethylation of RASSF1A, another TSG located immediately downstream of BLU, was detected in 20/27 (74%) of NPC tumors, no correlation between the hypermethylation of these two TSGs was observed (P=0.6334). In addition to methylation, homozygous deletion of BLU was found in 7/29 (24%) of tumors. Therefore, BLU is a stress-responsive gene, being disrupted in 83% (24/29) of NPC tumors by either epigenetic or genetic mechanisms. Our data are consistent with the interpretation that BLU is a TSG for NPC. PMID- 15122338 TI - Activation of diacylglycerol kinase alpha is required for VEGF-induced angiogenic signaling in vitro. AB - Vascular endothelial growth factor-A (VEGF-A) promotes angiogenesis by stimulating migration, proliferation and organization of endothelium, through the activation of signaling pathways involving Src tyrosine kinase. As we had previously shown that Src-mediated activation of diacylglycerol kinase-alpha (Dgk alpha) is required for hepatocytes growth factor-stimulated cell migration, we asked whether Dgk-alpha is involved in the transduction of angiogenic signaling. In PAE-KDR cells, an endothelial-derived cell line expressing VEGFR-2, VEGF-A165, stimulates the enzymatic activity of Dgk-alpha: activation is inhibited by R59949, an isoform-specific Dgk inhibitor, and is dependent on Src tyrosine kinase, with which Dgk-alpha forms a complex. Conversely in HUVEC, VEGF-A165 induced activation of Dgk is only partially sensitive to R59949, suggesting that also other isoforms may be activated, albeit still dependent on Src tyrosine kinase. Specific inhibition of Dgk-alpha, obtained in both cells by R59949 and in PAE-KDR by expression of Dgk-alpha dominant-negative mutant, impairs VEGF-A165 dependent chemotaxis, proliferation and in vitro angiogenesis. In addition, in HUVEC, specific downregulation of Dgk-alpha by siRNA impairs in vitro angiogenesis on matrigel, further suggesting the requirement for Dgk-alpha in angiogenic signaling in HUVEC. Thus, we propose that activation of Dgk-alpha generates a signal essential for both proliferative and migratory response to VEGF-A165, suggesting that it may constitute a novel pharmacological target for angiogenesis control. PMID- 15122339 TI - JNK activation is critical for Aplidin-induced apoptosis. AB - Aplidin is an antitumor drug that induces apoptosis and activates EGFR, Src, JNK and p38MAPK. Here, we show that Aplidin induces c-JUN, JUN B, JUN D, c-FOS, FRA-1 and FOS B genes of the activator-protein (AP)-1 family, and also p65/RELA, a major component of nuclear factor-kappa B (NF-kappaB). Concordantly, Aplidin increases AP-1 and NF-kappaB activity. c-FOS induction depends on EGFR, Src and JNK/p38MAPK. In contrast, induction of c-JUN does not require EGFR activity and p65/RELA induction is only partially dependent on these kinases. We used several genetically deficient cells to identify the critical target of Aplidin. Mouse embryo fibroblasts (MEFs) deficient for src, yes and fyn, and those lacking all p38MAPK isoforms displayed normal Aplidin sensitivity (IC50=12 nM). In contrast, MEFs lacking jnk1 and jnk2, which do not express any JNK isoform, were much less sensitive (IC50>500 nM). Furthermore, cells lacking c-jun or expressing a c-Jun protein in which JNK targets Ser(63/73) were mutated (c-JunAA) showed intermediate sensitivity (IC50=60 nM). Additionally, Aplidin has higher cytotoxic activity against proliferating than quiescent cells, which is reflected in higher JNK activation. We conclude that phosphorylation by JNK of c-Jun and additional substrate(s) is crucial for Aplidin activity. PMID- 15122340 TI - Nuclear FGF-2 facilitates cell survival in vitro and during establishment of metastases. AB - Nuclear-targeted high molecular weight 24 kDa fibroblast growth factor 2 (FGF-2) may induce specific cell functions through intracrine mechanisms. The role of nuclear FGF-2 on the metastatic potential of carcinoma cells was examined by conditional FGF-2 expression, which demonstrated that spontaneous metastasis in nude mice is a direct consequence of its expression. The lung colonizing capacities of fluorescent nuclear FGF-2-expressing cells following intravenous injection was also investigated. All cells reaching the lung extravasated as soon as 5 min following injection with similar in vivo behavior during the first 24 h. However, after 2 days, dramatic differences were observed between the FGF-2 and parental cells: most control cells underwent apoptosis, while the FGF-2-producing cells instigated a survival program and proliferated. Therefore, sustained apoptosis in vivo prevents growth of metastatic foci, while nuclear FGF-2 induction of a survival program is responsible for growth of the lung metastases. In vitro serum deprivation assays also established that 24 kDa FGF-2 expression improves carcinoma cell survival. This study provides both in vitro and in vivo evidence that the role of the nuclear 24 kDa FGF-2 isoform in carcinoma is the promotion of cell survival, thereby defining its association with poor prognosis in some human carcinomas. PMID- 15122341 TI - Polyomavirus tumorantigens have a profound effect on gene expression in mouse fibroblasts. AB - Polyomavirus (Py) large and small tumorantigens together are competent to induce S phase in growth-arrested mouse fibroblasts. The capacity of the large tumorantigen to bind the pocket proteins, pRB, p130 and p107, is important for the transactivation of DNA synthesis enzymes and the cyclins E and A, while the interference of small tumorantigen with protein phosphatase PP2A causes a destabilization of the cdk2 inhibitor p27, and thus leads to strong cyclin E- and cyclin A-dependent cdk2 activity. Py small tumorantigen, in addition, is able to transactivate cyclin A. Hence, this protein might have a much wider effect on gene expression in arrested mouse fibroblasts than hitherto suspected. This may have a profound part in the known capacity of Py to form tumors in mice. Therefore, it was interesting to gain an insight into the spectrum of transcriptional deregulation by Py tumorantigens. Accordingly, we performed microarray analysis of quiescent mouse fibroblasts in the absence and presence of small or large tumorantigen. We found that the viral proteins can induce or repress a great variety of genes beyond those involved in the S phase induction and DNA synthesis. The results of the microarray analysis were confirmed for selected genes by several methods, including real-time PCR. Interestingly, a mutation of the binding site for pocket proteins in case of LT and for PP2A in case of ST has a variable effect on the deregulation of genes by the viral proteins depending on the gene in question. In fact, some genes are transactivated by LT as well as ST completely independent of an interaction with their major cellular targets, pocket proteins and PP2A, respectively. PMID- 15122342 TI - Lupeol modulates NF-kappaB and PI3K/Akt pathways and inhibits skin cancer in CD-1 mice. AB - Chemoprevention has become an effective cancer control modality; however, the search for novel agent(s) for the armamentarium of cancer chemoprevention continues. We argue that agents capable for inhibition of promotion stage of tumorigenesis with the ability to intervene at several critical pathways in the tumorigenesis process will have greater advantage over other single-target agents. Lupeol, a triterpene, is the principal constituent of common fruit plants such as olive, mango, fig and medicinal herbs that have been used to treat skin aliments. Lupeol has been reported to possess a wide range of medicinal properties that include strong antioxidant, antimutagenic, anti-inflammatory and antiarthritic effects. In the present study, we show that Lupeol possesses antitumor-promoting effects in a mouse skin tumorigenesis model. We first determined the effect of topical application of Lupeol to CD-1 mouse against 12-O tetradecanoyl-phorbol-13-acetate (TPA)-induced conventional markers and other novel markers of skin tumor promotion. We found that topical application of Lupeol (1-2 mg/mouse) 30 min prior to TPA (3.2 nmol/mouse) application onto the skin of CD-1 mice afforded significant inhibition, in a time- and dose-dependent manner, against TPA-mediated increase in (i) skin edema and hyperplasia, (ii) epidermal ornithine decarboxylase (ODC) activity, and (iii) protein expression of ODC, cyclo-oxygenase-2 and nitric oxide synthase. As of the role of nuclear factor kappa B (NF-kappaB) and phosphatidyl inositol 3-kinase (PI3K)/Akt signaling in tumor promotion, we next determined the effect of topical application of Lupeol to mouse skin against these signaling pathways. We found that Lupeol treatment to mouse skin resulted in the inhibition of TPA-induced (i) activation of PI3K, (ii) phosphorylation of Akt at Thr(308), (iii) activation of NF-kappaB and IKKalpha, and (iv) degradation and phosphorylation of IkappaBalpha. The animals pretreated with Lupeol showed significantly reduced tumor incidence, lower tumor body burden and a significant delay in the latency period for tumor appearance. At the termination of the experiment at 28 weeks, 100% of the animals in TPA-treated group exhibited seven to eight tumors/mouse, whereas only 53% of the mice receiving Lupeol prior to TPA treatment exhibited one to three tumors/mouse. These results for the first time provide evidence that Lupeol possesses antiskin tumor-promoting effects in CD-1 mouse and inhibits conventional as well as novel biomarkers of tumor promotion. We suggest that Lupeol is an attractive antitumor-promoting agent that must be evaluated in tumor models other than skin carcinogenesis. PMID- 15122344 TI - Induction of S-phase arrest and p21 overexpression by a small molecule 2[[3-(2,3 dichlorophenoxy)propyl] amino]ethanol in correlation with activation of ERK. AB - We recently found that a small molecule 2[[3-(2,3 dichlorophenoxy)propyl]amino]ethanol (2,3-DCPE) could induce apoptosis and downregulate Bcl-XL expression in various cancer cells. Here, we found that 2,3 DCPE suppressed the proliferation of Bcl-XL-overexpressing cancer cells without inducing apoptosis. Subsequently, we found that 2,3-DCPE could induce S-phase arrest and upregulate p21 but not p27 at a time- and dose-dependent but p53 dispensable manner in DLD-1 human colon cancer cells. Activation of ERK was also detected after treatment with 2,3-DCPE. Moreover, p21 induction was dramatically attenuated by ERK inhibitors PD98059 and U0126. Induction of p21 and S-phase arrest and corresponding activation of ERK were also observed in ATM-defective cells, suggesting that 2,3-DCPE-induced these events were ATM-dispensable. Furthermore, ERK inhibitors dramatically attenuated 2,3-DCPE-induced S-phase arrest. Together, our data indicate that ERK activation correlated with the 2,3 DCPE-mediated induction of p21 expression and S-phase arrest. This finding may have implication for cancer therapy. PMID- 15122343 TI - Raf promotes human herpesvirus-8 (HHV-8/KSHV) infection. AB - Human herpesvirus-8 (HHV-8/KSHV) is etiologically associated with Kaposi's sarcoma (KS) and other tumors. Constitutive activation of the mitogen-activated protein kinase (MAPK) signaling pathway has been associated with a variety of tumors, including AIDS-related KS. The oncoprotein Raf is situated at a pivotal position in regulating the MAPK pathway. Hence, we analysed the effect of oncoprotein Raf on HHV-8 infectious entry into target cells. Here we report Raf expression to significantly enhance HHV-8 infection of target cells. These findings implicate a role for Raf not only in the infectious entry of HHV-8 but also in modulating KS pathogenesis. PMID- 15122345 TI - The sigma 70 subunit of RNA polymerase mediates a promoter-proximal pause at the lac promoter. AB - The sigma(70) subunit of RNA polymerase plays an essential role in transcription initiation. In addition, sigma(70) has a critical regulatory role during transcription elongation at the bacteriophage lambda late promoter, lambda P(R'). At this promoter, sigma(70) mediates a pause in early elongation through contact with a DNA sequence element in the initially transcribed region that resembles a promoter -10 element. Here we provide evidence that sigma(70) also mediates a pause in early elongation at the lac promoter (plac). Like that at lambda P(R'), the pause at plac is facilitated by a sequence element in the initially transcribed region that resembles a promoter -10 element. Using biophysical analysis, we demonstrate that the pause-inducing sequence element at plac stabilizes the interaction between sigma(70) and the remainder of the transcription elongation complex. Bioinformatic analysis suggests that promoter proximal sigma(70)-dependent pauses may play a role in the regulation of many bacterial promoters. PMID- 15122346 TI - The sigma 70 subunit of RNA polymerase induces lacUV5 promoter-proximal pausing of transcription. AB - The sigma(70) subunit of Escherichia coli RNA polymerase (RNAP) is a transcription initiation factor that can also be associated with RNAP during elongation. We provide biochemical evidence that sigma(70) induces a transcription pause at the lacUV5 promoter after RNAP has synthesized a 17 nucleotide transcript. The sigma(70)-dependent pausing requires an interaction between sigma(70) and a part of the lac repressor operator sequence resembling a promoter -10 consensus. The polysaccharide heparin triggers the release of sigma(70) from the paused complexes, supporting the view that during the transition from initiation to elongation the interactions between sigma(70) and core RNAP are weakened. We propose that the binding and retention of sigma(70) in elongation complexes are stabilized by its ability to form contacts with DNA of the transcription bubble. In addition, we suggest that the sigma(70) subunit in the elongation complex may provide a target for regulation of gene expression. PMID- 15122347 TI - Assembly of endocytic machinery around individual influenza viruses during viral entry. AB - Most viruses enter cells via receptor-mediated endocytosis. However, the entry mechanisms used by many of them remain unclear. Also largely unknown is the way in which viruses are targeted to cellular endocytic machinery. We have studied the entry mechanisms of influenza viruses by tracking the interaction of single viruses with cellular endocytic structures in real time using fluorescence microscopy. Our results show that influenza can exploit clathrin-mediated and clathrin- and caveolin-independent endocytic pathways in parallel, both pathways leading to viral fusion with similar efficiency. Remarkably, viruses taking the clathrin-mediated pathway enter cells via the de novo formation of clathrin coated pits (CCPs) at viral-binding sites. CCP formation at these sites is much faster than elsewhere on the cell surface, suggesting a virus-induced CCP formation mechanism that may be commonly exploited by many other types of viruses. PMID- 15122348 TI - Oxygen sensing by HIF hydroxylases. PMID- 15122349 TI - RAS and RHO GTPases in G1-phase cell-cycle regulation. PMID- 15122350 TI - A means to a DNA end: the many roles of Ku. PMID- 15122351 TI - Signals that control plant vascular cell differentiation. PMID- 15122352 TI - Shaping the nuclear action of NF-kappaB. PMID- 15122353 TI - Regulatory diversity among metazoan co-activator complexes. PMID- 15122354 TI - Actin up in the nucleus. PMID- 15122355 TI - Photosensitized inactivation of microorganisms. AB - Despite major advances in medicine in the last 100 years, microbiologically-based diseases continue to present enormous global health problems. New approaches that are effective, affordable and widely applicable and that are not susceptible to resistance are urgently needed. The photodynamic approach is known to meet at least some of these criteria and, with the creation and testing of new photosensitisers, may develop to meet all of them. The approach, involving the combination of light and a photosensitising drug, is currently being applied to the treatment of diseases caused by bacteria, yeasts, viruses and parasites, as well as to sterilisation of blood and other products. PMID- 15122356 TI - Photoinactivation of viruses. AB - Although the photodynamic effect was demonstrated against viral targets more than seventy years ago, the use of photosensitisers as antivirals in vivo has been slow in gaining acceptance. From a clinical viewpoint, this may be due to the pronounced side effects produced in several cases of the phototreatment of herpes genitalis in the early 1970s, the unfortunate patients presenting with post treatment Bowen's disease. Currently, the clinical use of photosensitisers in this field is limited to the treatment of laryngeal papillomata. However, considerable progress has been made in the photodynamic disinfection of blood products. Photoantivirals have traditionally been targeted at viral nucleic acid, in many cases via an intercalative mechanism. However, given the potential for deleterious sequelae associated with this route, the design of new photosensitisers should encourage alternative targets, such as viral enzymes or the cell envelope (where this exists). Targeting is obviously determined by the chemistry of the photosensitiser employed and there are many different structural types available. The chemistry, photochemistry and cellular effects of the various agents are discussed, along with future prospects for this exciting area of medicine. PMID- 15122357 TI - Lethal photosensitisation of oral bacteria and its potential application in the photodynamic therapy of oral infections. AB - Chemical antibacterial agents are increasingly being used in prophylactic and therapeutic regimes for dental plaque-related diseases, which are among the most common human infections. As these agents are difficult to maintain at a therapeutic concentration in the oral cavity and can be rendered ineffective by resistance development in the target organisms, there is a need to develop alternative antimicrobial approaches. Bacteria and other microbes can be sensitised to light through prior treatment with a chemical photosensitising agent. Lethal photosensitisation of a wide range of bacteria responsible for caries, periodontal diseases and root canal infections has been demonstrated using red light in conjunction with a number of photosensitisers, including Toluidine Blue, phthalocyanines and chlorins. The advantages of this approach are that bacteria can be eradicated in very short periods of time (seconds or minutes), resistance development in the target bacteria is unlikely and damage to adjacent host tissues and disruption of the normal microflora can be avoided. This approach may be a useful alternative to antibiotics and antiseptics in eliminating cariogenic and periodontopathogenic bacteria from disease lesions and for the disinfection of root canals. Not only would this be of benefit for the treatment of these diseases but, by replacing the antimicrobial agents that are currently used for such purposes, it would help to conserve our dwindling supply of antimicrobial agents that are effective in the treatment of serious systemic infections. PMID- 15122358 TI - In vitro effect of 5-aminolaevulinic acid plus visible light on Candida albicans. AB - Photodynamic therapy, currently used as an alternative technique for the treatment of superficial non-melanoma skin cancers, has been employed in vitro to kill different species of microorganisms. Here the development of Candida albicans colonies has been measured after application of 5-aminolaevulinic acid (ALA) plus visible light (VIS) irradiation. C. albicans suspensions (10 colony forming units microl(-1)) have been prepared. For the experiment 30 microl of suspension have been incubated in the dark for 3 h, with increasing concentrations of ALA (125, 250, 300, 350, 400, 450, 500, 550, 600, 750, 1000 mg ml(-1)) and then irradiated with a fixed dose (40 J cm(-2)) of VIS. Immediately after the irradiative session, the C. albicans suspensions were disseminated on dishes containing a Sabouraud agar + CAF medium and cultured in the dark at 27 degree C; after 48 h colony development has been measured. In the same way four controls have been prepared: (i)C. albicans suspensions not treated with ALA-PDT; (ii)C. albicans suspensions incubated with increasing ALA concentrations without VIS; (iii)C. albicans suspensions irradiated with 40 J cm(-2) of VIS without ALA; (iv)C. albicans suspensions irradiated immediately after the addition of increasing concentrations of ALA without the 3 h incubation. Colonies treated with ALA-PDT have been studied with electron microscopy (E.M.). It was found that: (i) none of the controls prepared modified the development of C. albicans colonies; (ii) ALA plus VIS inhibited C. albicans growth in a concentration dependent way: up to 250 mg ml of ALA concentrations did not affect C. albicans cells, 300 mg ml(-1) induced a 50% reduction in the number of colonies, a complete inhibition started from concentrations of 600 mg ml(-1); (iii) after ALA PDT E.M. showed modifications of the cell membranes. From the results it is concluded ALA plus VIS light is able to kill C. albicans colonies, at least in vitro. Although other pharmacological approaches are available, further studies could show that PDT is a potential treatment for candidosis. PMID- 15122359 TI - Mechanistic aspects of Escherichia coli photodynamic inactivation by cationic tetra-meso(N-methylpyridyl)porphine. AB - The mechanistic aspects of Escherichia coli photodynamic inactivation (PDI) have been studied in bacteria expressing the reporter protein GFP, following transfection with wild type pGFP plasmid and treatment with the hydrophilic cationic sensitizer tetra-meso(N-methyl-4-pyridyl)porphine tetratosylate (TMPyP). Cell survival and morphology during PDI were correlated with plasmid-GFP degradation in comparison to DNA and RNA strand-breaks, while photobleaching of the GFP chromophore was used to monitor protein photodamage. Singlet oxygen generated upon TMPyP photoactivation interacted with target nucleic acid polymers in a drug-and light-dose dependent manner. The hierarchy and cascade of the photodamage was in the order: genomic-DNA > total RNA > plasmid-DNA, as revealed by specific extraction and agarose electrophoresis. The notable resistance of the plasmid DNA in comparison to genomic DNA has implications for PDI of antibiotic resistant bacteria. Re-growth of the treated cells in fresh medium showed structural features of an SOS response. Under these conditions, DNA repair machinery was initiated by typical alignment of DNA-protein co-aggregates accompanied by lateral assembly of ribosomes, apart from damaged DNA-arrays, as depicted by electron microscopy. GFP-TMPyP interactions were demonstrated by double green and red fluorescence on electrophoresis plates analyzed by spectral imaging. Photobleaching measurements revealed specific GFP photodamage directly related to PDI of the E. coli. The kinetics of both the GFP photobleaching and the K(+) efflux, representing photodamage to cytosolic proteins and membrane damage, respectively, were found to be similar. The survival curves were correlated to chromosomal degradation and ultrastructural damage. We conclude that TMPyP-dependent PDI of E. coli is primarily dependent on genomic DNA photodamage rather than on protein or membrane malfunctions. PMID- 15122360 TI - ALA induced photodynamic effects on gram positive and negative bacteria. AB - In the present study we examined the production of high amounts of porphyrins upon induction by delta-aminolevulinic acid (ALA) in 9 bacterial strains. This was performed by solely inducing the porphyrin biosynthesis pathway. Four of the strains were Gram positive bacteria and five were Gram negative strains. All strains, except Streptococcus faecalis, produced porphyrins when incubated in PBS with 0.38 mM ALA for 4 h. Excess porphyrin production was excreted to the medium. Gram positive bacteria exhibited fluorescent emission peaks at 622 nm for the endogenous and 617 nm for the excreted porphyrins. Gram negative bacteria exhibited a 630 nm emission peak for the endogenous and a 615 nm emission peak for the excreted extracellular porphyrins. Upon illumination of the ALA induced Staphylococcal strains with 407-420 nm blue light, a decrease of five orders of magnitude was demonstrated with a light dose of 50 J cm(-2). Total eradication of the Staphylococcal strains could be achieved with a 100 J cm(-2) dose, which resulted in a decrease in viability of seven orders of magnitude. The viability of all the induced Gram negative strains and B. cereus decreased by one or two orders of magnitude upon illumination with 50 and 100 J cm(-2), respectively. This difference in the photoinactivation rate was found to be due to the distribution and amounts of the various porphyrins in the bacterial strains. The predominant porphyrin in the Staphylococcal strains was coproporphyrin (68.3 74.6%). In the Gram negative strains there was no predominant porphyrin and the porphyrins found were mostly 5-carboxyporphyrin, uroporphyrin, 7- carboxyporphyrin, coproporphyrin and protoporphyrin. In the B. cereus(Gram positive) strain the predominant porphyrin was uroporphyrin (75.8%). Although the total production of porphyrins in the Gram negative bacteria was higher than in the Staphylococcal strains, the amount of coproporphyrin produced by the latter was twice to three times higher than in the Gram negative strains. The extracellular excreted porphyrins did not contribute to the photoinactivation in any of the tested strains. Significant decreases in the Na(+) and K(+) content were detected in induced S. aureus after illumination while only small changes were observed in E. coli B. The green fluorescent protein within the cytoplasm of induced E. coli strains was only partially disrupted (by 60% only). These results indicate a partial yield of the effects generated by (1)O(2) radicals resulting from the photoinactivation of Gram negative bacteria and a successful generation of the same effects in the Staphylococcal strains. PMID- 15122362 TI - Targeted photodynamic therapy of established soft-tissue infections in mice. AB - The worldwide rise in antibiotic resistance necessitates the development of novel antimicrobial strategies. Although many workers have used photodynamic therapy (PDT) to kill bacteria in vitro, the use of this approach has seldom been reported in vivo in animal models of infection. We have previously described the first use of PDT to treat excisional wound infections by Gram-(-) bacteria in living mice. However, these infected wound models involved a short timespan between infection (30 min) and treatment by PDT. We now report on the use of PDT to treat an established soft-tissue infection in mice. We used Staphylococcus aureus stably transformed with a Photorhabdus luminescenslux operon (luxABCDE) that was genetically modified to be functional in Gram-(+) bacteria. These engineered bacteria emitted bioluminescence, allowing the progress of the infection to be monitored in both space and time with a low light imaging charge coupled device (CCD) camera. One million cells were injected into one or both thigh muscles of mice that had previously been rendered neutropenic by cyclophosphamide administration. Twenty-four hours later, the bacteria had multiplied more than one hundredfold; poly-L-lysine chlorin e6 conjugate or free chlorin e6 was injected into one area of infected muscle and imaged with the CCD camera. Thirty minutes later, red light from a diode laser was delivered as a surface spot or by interstitial fiber into the infection. There was a light dose dependent loss of bioluminescence (to <5% of that seen in control infections) not seen in untreated infections or those treated with light alone, but in some cases, the infection recurred. Treatment with conjugate alone led to a lesser reduction in bioluminescence. Infections treated with free chlorin e6 responded less well and the infection subsequently increased over the succeeding days, probably due to PDT-mediated tissue damage. PDT-treated infected legs healed better than legs with untreated infections. This data shows that PDT may have applications in drug-resistant soft-tissue infections. PMID- 15122363 TI - Characterization of RS29, a blue-green proteorhodopsin variant from the Red Sea. AB - Using structural modeling comparisons and mutagenesis, amino acid residue 105 was found to function as a spectral tuning switch in marine proteorhodopsins (PR). Changes at this position account for most of the spectral difference between blue absorbing PRs (B-PRs), and green-absorbing PRs (G-PRs). Here we analyzed a Red Sea variant (RS29) from a new family of PRs that is composed of G-PR type variants that possess glutamine instead of leucine at position 105 like in B-PRs. The absorption spectrum as well as photocycle of RS29 variant were measured and compared to point-mutated 'position 105' PRs. Unexpectedly, the absorption maximum of RS29 was 515 nm, a smaller blue shift compared to the 498 nm maximum of G-PR_L105Q. We found that two additional residues at positions 65 and 70 each contribute a small red shift to the absorption spectrum of G-PR and therefore appear to account for the intermediate absorption maximum of RS29 by their opposing influences on the spectrum. Our results show that in addition to the retinal pocket position 105 determinant, other residues predicted to be outside the retinal pocket fine-tune the absorption spectra of marine PRs. The RS29 photochemical reaction cycle was found to be 2 orders of magnitude slower than that of G-PR with a t(1/2) of >600 ms. This result raises the possibility of regulatory (i.e. sensory) rather than energy harvesting functions for some members of the PR family. PMID- 15122364 TI - Catechin and hydroxybenzhydrols as models for the environmental photochemistry of tannins and lignins. AB - The photochemistry of several model plant-derived compounds has been studied in aqueous solution. In particular, the reactions of catechin as a model tannin and methoxy-substituted hydroxybenzhydrols as model lignin functionalities were investigated. Tannins and lignins constitute a significant portion of the humic substances in aquatic systems, which are themselves the main component of dissolved organic matter thought to be responsible for the absorption and attenuation of light in these environments. Catechin (1) was found to undergo a reversible photoisomerization reaction to give epicatechin (2). Such a reaction is an explicit example of a photon absorbing process that enables catechin (1) and its derivatives to act as natural sunscreens by attenuating light energy through non-destructive reactions. The methoxy-substituted hydroxybenzhydrols were found to undergo photosolvolysis reactions via efficient generation of quinone methide intermediates. The intermediate quinone methides were observed to be longer lived, and thus more stable, than previously studied hydroxybenzhydrol derivatives. The meta-hydroxybenzhydrol isomer (5) was found to undergo additional chemistry leading to the production of a ring-closed fluorene from the quinone methide intermediate. PMID- 15122361 TI - Photodynamic therapy: a new antimicrobial approach to infectious disease? AB - Photodynamic therapy (PDT) employs a non-toxic dye, termed a photosensitizer (PS), and low intensity visible light which, in the presence of oxygen, combine to produce cytotoxic species. PDT has the advantage of dual selectivity, in that the PS can be targeted to its destination cell or tissue and, in addition, the illumination can be spatially directed to the lesion. PDT has previously been used to kill pathogenic microorganisms in vitro, but its use to treat infections in animal models or patients has not, as yet, been much developed. It is known that Gram-(-) bacteria are resistant to PDT with many commonly used PS that will readily lead to phototoxicity in Gram-(+) species, and that PS bearing a cationic charge or the use of agents that increase the permeability of the outer membrane will increase the efficacy of killing Gram-(-) organisms. All the available evidence suggests that multi-antibiotic resistant strains are as easily killed by PDT as naive strains, and that bacteria will not readily develop resistance to PDT. Treatment of localized infections with PDT requires selectivity of the PS for microbes over host cells, delivery of the PS into the infected area and the ability to effectively illuminate the lesion. Recently, there have been reports of PDT used to treat infections in selected animal models and some clinical trials: mainly for viral lesions, but also for acne, gastric infection by Helicobacter pylori and brain abcesses. Possible future clinical applications include infections in wounds and burns, rapidly spreading and intractable soft tissue infections and abscesses, infections in body cavities such as the mouth, ear, nasal sinus, bladder and stomach, and surface infections of the cornea and skin. PMID- 15122365 TI - Pressure dependence of the dual luminescence of twisting molecules. The case of substituted 2,3-naphthalimides. AB - The effect of high pressure (up to 5 kbar) has been studied for triacetin solutions of 2-phenyl-2,3-dihydro-1H-benzo[f]isoindole-1,3dione 1 (N-phenyl-2,3 naphthalimide) and its 3-fluorophenyl- (2), 4-carbethoxyphenyl(4) and 4 methoxyphenyl (5) derivatives which all show dual fluorescence. When the N-phenyl group is unsubstituted (compound 1) or substituted with electron-attracting groups (2 and 4), the increase of pressure over the solution decreases slightly the emission at the long-wavelengths (LW) and increases dramatically the intensity of the short-wavelength (SW) fluorescence. Plotting the logarithm of the SW/LW fluorescence quantum yield ratio for compounds, 1,2 and 4 versus the logarithm of the viscosity of the medium shows a substantial increase of this ratio which corresponds mainly to the increase of the SW emission intensity, the effect on the LW emission being only moderate. As the pressure is increased, the rotation of the N-phenyl group of compound 1 is progressively hindered and the prevailing emission comes from a state which has the same geometry as the ground state (in which the planes of the two moieties of the molecule form an angle close to 60 degree). The effect is different when an electron-donating methoxy group is attached in the para position to the N-phenyl ring, compound 5, as mainly the LW fluorescence intensity increases with pressure. For this molecule which has an electron-donating p-substituent on the N-phenyl ring, the two moieties of the ground state molecule have a more planar geometry (43 degree angle) and the LW fluorescence appears to originate from an intramolecular charge transfer state the fluorescence of which increases with pressure. A three-level reaction scheme is proposed to account for the observed kinetics. In all cases, the viscosity of the medium is found to be the main factor which induces the changes in the fluorescence spectra, and the deceleration of the non-radiative deactivation from the SW* excited state is responsible for these modifications whether a reversible process between the two emitting SW* and LW* states is observed (as for compounds 2 and 4) or not (as for compound 1). PMID- 15122366 TI - Effect of SiO2 and zeolite surfaces on the excited triplet state of benzophenone, BT; a spectroscopic and kinetic study. AB - Laser flash photolysis has been used to study the triplet excited state of benzophenone B(T), on various surfaces, SiO(2), zeolites NaY, KY, NaX and KX, and in rigid media at room temperature, polyethylene and polymethylmethacrylate. The studies point to similarities of the spectroscopy and kinetics of B(T) in fluid solution, in a solid matrix (polymers) and on a SiO(2) surface. However, stark differences are observed for B(T) in zeolites where the absorption spectrum mimics that of the protonated ketone, and the reactivities of B(T) with C(6)H(12) and CH(3)OH are an order of magnitude smaller than those in liquid C(6)H(12) and CH(3)OH. Inclusion of ammonia, which blocks acidic sites in the zeolite, produces a triplet spectrum which is similar to that in polar solution. The reactivity of the triplet with ammonia in a zeolite is also comparable to that observed for this reaction in polar solution. These data are discussed in terms of the interaction of benzophenone with acidic sites in the zeolites, and to restrictions placed on the reactants in the zeolite cages. The blocking of the zeolite acidic sites by ammonia produces spectral and kinetic data (reactivity with NH(3)) of the triplet that are comparable to those observed in solution. This is one of the few cases where zeolites inhibit rather than promote reactions of a solute adsorbed in them. PMID- 15122367 TI - Photosensitized oxidation of phenyl and tert-butyl sulfides. AB - The photosensitized oxygenation of diphenyl (1), di-tert-butyl (2) and phenyl tert-butyl sulfide (3) was studied. Bimolecular rate constants of singlet oxygen quenching are low (1 to 5 x 10(4) M(-1)s(-1)) since the sulfides are poor nucleophiles due to sterical hindrance (2, 3) or the HOMO on the sulfur atom being a less accessible p(z) orbital (1). The quenching is mainly physical, but chemical reaction leading to sulfoxides also takes place in methanol and, to a lower degree, in acetonitrile. Catalysis by carboxylic acids considerably enhances the rate of sulfoxidation. Inefficiency in the chemical reaction is again due to the poor nucleophilicity of the sulfides, which limits oxygen transfer by electrophilic intermediates such as the protonated persulfoxide. PMID- 15122368 TI - [Work ability of subjects with chronic musculoskeletal disorders]. AB - OBJECTIVE: To evaluate the impact of personal, clinical, and occupational aspects on work ability of workers with and without work-related musculoskeletal disorders using the Work Ability Index. METHODS: There were participating 127 workers of industrial production lines of a medium-size multinational company. An approved version of the Work Ability Index, a questionnaire developed by Finland's Institute of Occupational Health, was used. Pain scale was also applied. A descriptive analysis was carried out using the Chi-square test and it was also performed a logistic regression analysis. RESULTS: A significant association was identified between the Work Ability Index and all personal, clinical and occupational aspects. Regression analysis showed that pain and sick leave together accounted for 59% of poor work ability. CONCLUSIONS: Different levels of pain severity were associated with distinct as well as equivalent levels of work ability loss. The results suggest that either pain reports were consistent or being both pain and work ability self-reported they therefore reflect the same perception mechanisms. Future studies might contribute to further understanding the trends found. PMID- 15122369 TI - [Prevalence of sedentarism and its associated factors among urban adolescents]. AB - OBJECTIVE: To evaluate the prevalence and determinants of sedentarism among adolescents living in Pelotas, State of Rio Grande do Sul, Brazil. METHODS: A cross-sectional study was carried out in Pelotas, in 2002. An anonymous, self administered questionnaire was answered by a representative sample of 960 adolescents with ages ranging from 15 to 18 years old. Those who reported that their participation in some kind of physical activity amounted to less than 20 minutes a day with a frequency of less than 3 times a week were considered to have a sedentary lifestyle. Sociodemographic and behavioral variables were evaluated. The chi-squared test was used for comparisons between proportions. The Poisson regression was used for multivariate analysis with robust adjustment for variances. Corrections were made for study design effects. RESULTS: Interviews were held with 960 adolescents. Of these, 39% were classified as having a sedentary lifestyle. The prevalence of sedentarism was higher among girls than among boys (prevalence ratio of 2.45; 95% CI 2.06-2.95). The adolescents from lower social levels had the highest percentage of sedentarism (prevalence ratio of 1.35; 95% CI 1.06-1.72). There was an inverse relationship between the adolescent's and the mother's schooling and sedentarism. After adjusting for confounding factors, there was also a positive association between sedentarism and minor psychiatric disturbances, and in relation to sexual activity. CONCLUSIONS: Higher prevalences of sedentarism are associated with girls, low social class, low levels of schooling and being the child of a mother with a low level of schooling. PMID- 15122370 TI - [Short version of the "job stress scale": a Portuguese-language adaptation]. AB - OBJECTIVE: To describe the adaptation to Portuguese of the short version of the "job stress scale", originally in English. METHODS: We evaluate six aspects of equivalence between the original scale and the Portuguese version: conceptual, semantic, operational, item, measurement, and functional equivalences. A reliability test-retest study was conducted with 94 selected subjects. RESULTS: Reproducibility (interclass correlation coefficients) for the 'demand', 'control', and 'social support' dimensions of the scale was estimated at 0.88, 0.87, and 0.85, respectively. Internal consistency (Cronbach's alpha) estimates for these same dimensions were 0.79, 0.67, and 0.85, respectively. CONCLUSIONS: Our results suggest that the adaptation of the scale was successful, and indicate that its use in the sociocultural context of the studied population (Pro-Saude survey) is appropriate. PMID- 15122371 TI - [Exclusive breastfeeding among working women with free daycare available at workplace]. AB - OBJECTIVE: To investigate factors related to the decision of exclusive breastfeeding, and the planned and the actual duration among working women with free daycare available at workplace. METHODS: A qualitative study was conducted comparing a group of 15 women exclusively breastfeeding their babies with a similar group of women whose babies were already being fed with other food besides maternal milk at the time they started attending a daycare center. Semi structured interviews and focus groups were carried out for data collection. RESULTS: The factors related to the decision of breastfeeding and maintaining it when women went back to work were: the desire to breastfeed based on the importance women of both groups as well as their husbands and significant others attributed to it. The duration of exclusive breastfeeding was mainly associated to the baby's pediatrician counseling, which differed in each group. CONCLUSIONS: The availability of free daycare center at the work place seems an important aspect to breastfeeding maintenance after women go back to work, especially regarding exclusive breastfeeding. The duration of exclusive breastfeeding was related to the information received before and during pregnancy, and also in the postpartum. Women who have exclusively breastfed for almost six months believed the longer they breastfeed the better to their babies' health, while other women believed that three months of exclusive breastfeeding would be enough. PMID- 15122372 TI - [Infant mortality evolution in the metropolitan region of Sao Paulo (Brazil), 1980-2000]. AB - OBJECTIVE: To analyze IMR evolution in the Sao Paulo Metropolitan Area (SPMA) between 1980 and 2000, in terms of spatial, age and, causal differences. METHODS: SPMA municipalities were divided into 5 groups, based on their 1980 IMRs: 90 per thousand lb (Group 1); 70-89 per thousand lb (Group 2); 50-69 per thousand lb (Group 3); <50 per thousand lb (Group 4). Group 5 comprised the municipality of Sao Paulo itself (IMR=51 per thousand lb). The analysis of trends was carried out using exponential regression models. RESULTS: IMR and its components showed a statistically significant decrease (p<0.05), with coefficients of determination between 66 and 98%, indicating goodness of fit of the exponential model to all the time series analyzed. SPMA IMR fell 69.4%, from 55.2 to 16.9 per thousand lb, and Groups 1-5 showed reductions of 83.9%, 76.2%, 71.3%, 58.7%, and 68.8%, indicating that the groups with highest IMRs also showed the greatest reductions during the studied period. CONCLUSIONS: IMRs were homogenized at around 18 lb in all municipality groups in the SPMA. One-half of all deaths were concentrated within the first week of life, and were due primarily to conditions originating in the perinatal period, indicating that greater care during the pre- and post delivery periods will be required if the IMR in the SPMA is to descend to levels compatible with those of developed countries. PMID- 15122373 TI - [Prevalence and associated factors to hepatitis C in hemodialysis patients in Brazil]. AB - OBJECTIVE: The prevalence of hepatitis C virus (HCV) is higher in patients on hemodialysis than in the general population, probably due to greater exposure to risk situations. The purposes of the study were to determine anti-HCV antibodies prevalence among hemodialysis patients and dialysis clinics and patients factors associated with HCV transmission. METHODS: A cross-sectional study was conducted in 752 hemodialysis patients in all 12 dialysis clinics of Fortaleza, Brazil, and were screened using third generation ELISA. Sociodemographic, clinical, and epidemiological data of 663 patients were collected through interviews. Nosocomial factors were assessed using a specific questionnaire tool. Statistical analysis was conducted using Student's t test, odds ratio and multivariate analysis. RESULTS: The prevalence of anti-HCV was 52% (390/746; ranged from 6% to 72%). The anti-HCV positivity was higher in patients who had previous peritoneal dialysis (OR=1.76; 95% CI 1.12-2.76) and blood transfusion (OR=2.75; 95% CI 1.25 6.03). Dialysis age has been associated with anti-HCV positivity (OR=1.47; 95% CI 1.35-1.61). Clinics practices associated with anti-HCV positivity were: previous preparing of heparin (OR=2.92; 95% CI 1.23-6.92), failure in gloves use or change (OR=5.73; 95% CI 1.75-18.72), unsatisfactory dialysis machine disinfection (OR=2.79; 95% CI 1.57-4.96), and patient isolation in dialysis room (OR=0.18; 95% CI 0.05-0.61). CONCLUSIONS: The results show high anti-HCV prevalence among hemodialysis patients and the association of nosocomial factors with new HCV infection cases. PMID- 15122374 TI - [Contraceptive measures and HIV transmission protection among women with HIV/AIDS]. AB - OBJECTIVE: Sexual intercourse is currently the route of transmission among women that has most contributed to the feminization of the HIV/AIDS epidemic. As an ongoing effort to establish more appropriate standards for health counseling, the study's purpose was to investigate the use of contraceptive methods that would also prevent HIV/AIDS women against disease transmission. METHODS: An exploratory study was developed in an outpatient clinic of a public university hospital, a reference center of HIV/AIDS patients in the mid-south region of the state of Sao Paulo, Brazil, during a 5-month-period (2000 and 2001). The study was carried out in 73 HIV/AIDS women. Data were collected using a semi-structured questionnaire exploring subjects' sociodemographics, contraception method used and HIV status of their sex partners. A descriptive data analysis was performed and the contents of open answers were grouped into themes. Fischer's exact test was applied for analyzing some variables at a 5% significance level. Content analysis was carried out according to Bardin's proposal.(2) RESULTS: Most women at reproductive age were married and had been infected almost exclusively through heterosexual contact. Of them, 35.4% reported having an HIV discordant partner and 13.7% used inadequate contraceptive methods that failed to protect them against HIV transmission. CONCLUSIONS: The study results call for the need of continuous education on safer sex among HIV/AIDS women to empower them to discuss with their partners alternative options of exercising their sexuality and to raise awareness on their contraceptive choices in a way to protect their own health, their partner's and even their unborn offspring's health. PMID- 15122375 TI - [Prevalence of low visual acuity in public school's students from Brazil]. AB - OBJECTIVE: Low visual acuity (VA) is an important public health problem due to its high prevalence and because it needs early diagnosis in order to prevent damage in childhood development and apprenticeship. To describe and analyze low visual acuity (VA) prevalence among school children. METHODS: Once performed the VA test to 1st and 4th grades primary school children data were analyzed by separating students according to sex, school grade, wearing of glasses, residence area and level of access to the supplementary medical assistance (SMA). RESULTS: The total of 9,640 students was evaluated during the year of 2000 and they presented a prevalence of low VA of 13.1% (CI 12.5-13.8%). There was a statistical significant lower prevalence in males (11.5%) compared to females (14.9%) - (PR=0.77). There was a statistical significant higher prevalence in 1st grade students (14.1%) compared to 4th grade (11.5%) - (PR=1.22). There was also a statistical significant lower prevalence for those who were not wearing glasses (12.1%) compared to those who were using glasses (42.0%) - (PR=0.29). Concerning to residence areas, Cajuru neighborhood had the lower prevalence of low VA (1.8%) and Vila Sabia neighborhood had the higher prevalence (32.4%), and a positive correlation, according to residence area, between the proportion of people with access to the Supplementary Medical Assistance and the proportion of children wearing glasses was found (r=0.64, p<0.001). CONCLUSIONS: The low VA high prevalence shows lack in early diagnosis and continuity of assistance pointing out to the urgent need of implementation in visual health public. PMID- 15122376 TI - [Productivity of Aedes albopictus' breeding containers in Paraiba Valley, Brazil]. AB - OBJECTIVE: To estimate the potential productivity of artificial, permanent, and natural breeding containers of Aedes albopictus, potential vector species of dengue. METHODS: Three locations were selected for this study: a) warehouse of disposable material goods in the urban area of Pindamonhangaba, state of Sao Paulo, Brazil; b) a small leisure farmland in Tremembe, state of Sao Paulo; and c) residual forest in the periurban area of Pindamonhangaba. All breeding containers were assessed and classified according to their water volume ( small (up to 1 liter), medium (over 1 but up to 10 liters), and large (over 10 liters), and type (artificial, natural, and permanent). Collections were carried out fortnightly for a period of 12 months, and a sample of fourth-instar larvae and pupae were collected from each container. Statistical analysis was performed using the Kriskal-Wallis test, t-Student test, and emergence estimate. RESULTS: The statistical analyses and emergence estimate (E) showed that artificial large size containers and permanent medium container were on average the most productive, resulting 2.8 females a day, each. Natural small and medium containers produced on average 0.5 and 0.6 females respectively. CONCLUSIONS: There is a need for further studies on the potential productivity of containers not only of Ae. albopictus but also of Aedes aegypti, the main vector of dengue, which could contribute to better assessments of population densities in surveillance and control programs. PMID- 15122377 TI - Entomological aspects of Chagas' disease transmission in the domestic habitat, Argentina. AB - OBJECTIVE: To study the risk of Trypanosoma cruzi domestic transmission using an entomological index and to explore its relationship with household's characteristics and cultural aspects. METHODS: There were studied 158 households in an endemic area in Argentina. Each household was classified according to an entomological risk indicator (number of risky bites/human). A questionnaire was administered to evaluate risk factors among householders. RESULTS: Infested households showed a wide range of risk values (0 to 5 risky bites/human) with skewed distribution, a high frequency of lower values and few very high risk households. Of all collected Triatoma infestans, 44% had had human blood meals whereas 27% had had dogs or chickens blood meals. Having dogs and birds sharing room with humans increased the risk values. Tidy clean households had contributed significantly to lower risk values as a result of low vector density. The infested households showed a 24.3% correlation between time after insecticide application and the number of vectors. But there was no correlation between the time after insecticide application and T. infestans' infectivity. The statistical analysis showed a high correlation between current values of the entomological risk indicator and Trypanosoma cruzi seroprevalence in children. CONCLUSIONS: The risk of T. cruzi domestic transmission assessed using an entomological index show a correlation with children seroprevalence for Chagas' disease and householders' habits. PMID- 15122378 TI - [Level of knowledge of the compositions of analgesic medication containing aspirin]. AB - OBJECTIVE: To evaluate the knowledge of the generic designation, use and composition of analgesic medications containing aspirin. METHODS: A total of 124 interviews were carried out between December 1999 and January 2000, in two neighborhoods of the city of Porto Alegre, southern Brazil. The interview was held with the person who came to answer the door at each of the homes that was drawn. The data collection instruments comprised a set of five different pharmaceutical specialties containing acetylsalicylic acid, and an interview consisting of two open questions concerning the differences and similarities between the products. RESULTS: Three major knowledge-level groups were characterized on the basis of the information that the interviewees were able to provide. The group that was knowledgeable about the matter comprised 14 individuals (11%). The group with limited knowledge contained 61 people (49)%. Those who had no knowledge of the matter at all formed a group of 49 people (40%). CONCLUSIONS: Taking the results as a whole, they indicate that most people (about 90% of the sample investigated) are simply not aware of what the active substance is, even in pharmaceutical specialties that they use frequently. PMID- 15122379 TI - [Drug utilization in adults: prevalence and individuals determinants]. AB - OBJECTIVE: To study epidemiological patterns of drug utilization and its individual determinants and to classify drugs used into pharmacological groups. METHODS: In a population-based cross-sectional study, 3,182 subjects aged 20 years or more were selected from an urban area in Southern Brazil using a multi stage sampling design. Data were collected through home interviews using a structured questionnaire. Drug use in the previous 15 days was recorded. Crude analysis was performed using the Chi-square test for heterogeneity and trend and t-test for means' comparison. The adjusted analysis was carried out using Poisson regression. RESULTS: The overall prevalence of drug utilization was 65.9%. The highest prevalences of drug use after adjusting for confounding were seen among elderly women in the upper economic class who had ill health self-perception. Analgesics, anti-inflammatory and anti-hypertensive drugs were the most frequently used drugs. CONCLUSIONS: The prevalence of drug use was found to be higher than in other national and international studies. Studies focusing on individual determinants of drug utilization can identify those groups with higher use. This knowledge might encourage specific strategies to decrease drug use and lead to restrictive policies concerning drug prescription and sale. PMID- 15122380 TI - [Use of antimicrobial drugs in an urban population]. AB - OBJECTIVE: The emergence of multiresistant microorganisms has been a concerning matter worldwide in the last decades. Indiscriminate use of antibiotics has been associated to this phenomenon. The present study was designed to determine the pattern of antimicrobial drug use in an urban community. METHODS: A population based cross-sectional study was carried out and 6,145 subjects of all ages living in the urban area of Pelotas, Brazil, were interviewed on the use of antimicrobial drugs in the 30 days previous to the interview. RESULTS: The overall prevalence of antimicrobial drug use was 8%. It was higher for children under 4 years of age (14%; p<0.001), women (9%; p=0.004) and divorced subjects (10%; p=0.02). The clinical conditions most frequently associated with antimicrobial drug use were respiratory tract infections (50%), urinary tract infections (16%), and dental infections (9%). Penicillins (41%), sulphas (17%), and tetracycline (8%) were the most commonly used drugs. CONCLUSIONS: Abuse of last generation antimicrobial drugs, a concern of many experts, was not confirmed in this study. Antimicrobial drug use could be further reduced as respiratory illnesses, most frequently associated with their use, are often viral and many patients will not benefit from antimicrobial therapy. PMID- 15122381 TI - Are severe occlusal problems more frequent in permanent than deciduous dentition? AB - OBJECTIVE: To determine the prevalence and severity of occlusal problems in populations at the ages of deciduous and permanent dentition and to carry out a meta-analysis to estimate the weighted odds ratio for occlusal problems comparing both groups. METHODS: Data of a probabilistic sample (n=985) of schoolchildren aged 5 and 12 from an epidemiological study in the municipality of Sao Paulo, Brazil, were analyzed using univariate logistic regression (MLR). Results of cross-sectional study data published in the last 70 years were examined in the meta-analysis. RESULTS: The prevalence of occlusal problems increased from 49.0% (95% CI =47.4%-50.6%) in the deciduous dentition to 71.3% (95% CI =70.3%-72.3%) in the permanent dentition (p<0.001). Dentition was the only variable significantly associated to the severity of malocclusion (OR=1.87; 95% CI =1.43 2.45; p<0.001). The variables sex, type of school and ethnic group were not significant. The meta-analysis showed that a weighted OR of 1.95 (1.91; 1.98) when compared the second dentition period with deciduous and mixed dentition. CONCLUSIONS: In planning oral health services, some activities are indicated to reduce the proportion of moderate/severe malocclusion to levels that are socially more acceptable and economically sustainable. PMID- 15122382 TI - [Sexual and reproductive health of female inmates in Brazil]. AB - OBJECTIVE: To describe the sociodemographic profile and health problems of inmates in a women's prison. METHODS: A descriptive study was conducted from March to September 1997 in a women's prison in the state of Espirito Santo, Brazil. All inmates were invited to participate in the study. A total of 121 women aged more than 18 years were interviewed. A structured questionnaire was used to collect data on sociodemographics, clinical and criminal past history. Following the interview, subjects underwent clinical and gynecological examination. RESULTS: Of 121 women included in the study, the mean age was 30.2 years (SD 8.98) and the mean of schooling was 4.8 years (SD 3.50). All participants reported previous sexual activity, the mean age of the first sexual intercourse was 15.2 years (SD 2.55) ranging from 9 to 27 years. Previous STDs were reported by 28%; 12 (9.9%) were pregnant at the time. Teenage pregnancy was often reported. Most women reported no use of either any contraceptive methods or condoms. Tubal ligation was seen in 19.8% and abnormal Pap smear in 26.9%. CONCLUSIONS: Knowledge about health problems inside the prison system can contribute to promoting and increasing social rehabilitation. However, effective results could be seen only with collaboration between public health authorities and the prison system. PMID- 15122383 TI - [Correlates of smoking in pregnant women in six Brazilian cities]. AB - OBJECTIVE: To evaluate sociodemographic and lifestyle correlates of smoking in pregnant women sampled from hospitals. METHODS: A cross-sectional study was conducted in 5,539 pregnant women aged 20 or more who sought medical attention in prenatal clinics of affiliate hospitals of the Brazilian National Health System in the cities of Manaus, Fortaleza, Salvador, Rio de Janeiro, Sao Paulo, and Porto Alegre from 1991 to 1995. Interviews were conducted using a standardized questionnaire that covered sociodemographics and smoking habits before and during pregnancy. Current smoking was defined as smoking at least one cigarette/day, former smoking as reporting having smoked at least one cigarette/day but having quit, and never smoking as never having smoked one cigarette/day. RESULTS: Smoking during pregnancy was associated with lower education (OR=2.13; CI 95%: 1.76-2.57) and greater parity (OR=1.84; CI 95%: 1.53-2.21). Positive associations were also found with increased gestational age and alcohol consumption. No significant association was found with skin color or occupation status. A protective effect was observed for women married or living with a partner (OR=0.55 CI 95%: 0.42-0.72). Having Manaus' women as a reference, Porto Alegre's women showed the greatest risk for smoking in pregnancy (OR=5.00; CI 95%: 3.35 7.38), followed by Sao Paulo's (OR=3.42; CI 95%: 2.25-5.20), Rio de Janeiro (OR=2.53; CI 95%: 1.65-3.88) and Fortaleza's (OR=2.56; CI 95%: 1.74-3.78). CONCLUSIONS: The study findings are similar to those described in the literature regarding education, parity, and marital status. However, no association with skin color was seen in the multivariate analysis. Former smokers had sociodemographic characteristics more similar to non-smokers than former smokers. PMID- 15122384 TI - [Histories of fetal losses told by women: research qualitative study]. AB - OBJECTIVE: To recognize the significance of fetal loss for women who have experienced it, starting from an understanding of the pregnancy process, based on their reports. METHODS: This was a qualitative analysis study based on the histories of seven women who experienced fetal loss in the town of Aruja, State of Sao Paulo, between July 1998 and June 1999. The women were identified from the death certificates of stillborn infants born within the study period, which were obtained from the Civil Registry Office of Aruja. The methodological procedures involved the utilization of the techniques of oral history-taking to gather data and content analysis to evaluate the material collected. The interviews were recorded, fully transcribed and subsequently prepared for analysis. RESULTS: The findings were analyzed as two points: the circumstantial context of the pregnancy and the impact after the loss, with the adoption of specific thematic categories. The first of these encompassed the woman's perception of the pregnancy, her awareness of the coming of the new baby, health problems up to the time of the loss, and the health service attendance. The significance of the loss for the women in this study was made evident along three central lines: the loss of a part of herself, attribution of the fatality to divine intervention and changes in attitude towards life. The social support network for these women was built on two pillars: family and church. Support from the health services was practically nonexistent. Finally, they all expressed their will to live and the need to work, study and even have another pregnancy. CONCLUSIONS: There needs to be a change in general concepts in the mission to attend to such women. The attendance provided by the healthcare services needs to be humanized. The need for multiprofessional follow-up of healthcare service users who suffered fetal loss was very evident. The importance of a support network for women who have gone through this problem was also shown. PMID- 15122385 TI - [Use of psychotropics drugs among students: prevalence and associated social factors]. AB - OBJECTIVE: To determine the prevalence of the heavy use of drugs among elementary and high school students in a sample of public and private schools, and to identify associated demographic, psychological, cultural and social factors. METHODS: This report describes a cross-sectional study using an intention-type sampling technique that compared public schools in central and peripheral areas and private schools. An anonymous self-administered questionnaire was applied. The sample consisted of 2,287 elementary and high school students in the city of Campinas in 1998. Heavy use of drugs was defined as the use of drugs on 20 or more days during the 30 days preceding the survey (WHO, 1981). For the statistical analysis, polytomic logistic regression analysis (logit model) was utilized to identify factors that influenced this manner of using drugs. RESULTS: Heavy use of legal and illegal drugs was found as follows: alcohol (11.9%), tobacco (11.7%), marijuana (4.4%), solvents (1.8%), cocaine (1.4%), medications (1.1%) and ecstasy (0.7%). The heavy use of drugs was greatest among students at the city-center public school who had daytime jobs and studied in the evenings. These students were in the A and B socioeconomic classes and had had little religious education during childhood. CONCLUSIONS: Greater availability of cash and specific socialization patterns were identified as factors associated with the heavy use of drugs among students. PMID- 15122386 TI - [Heavy alcohol consumption and associated factors: a population-based study]. AB - OBJECTIVE: To determine the prevalence of heavy alcohol consumption and factors associated with it in a Brazilian adult population. METHODS: Cross-sectional population-based study including 2,177 adults (aged 20 to 69), living in the urban area of the municipality of Pelotas, Rio Grande do Sul State, Brazil. The sample was selected in multiple stages. Heavy alcohol consumption was defined as above 30g/day. The adjusted analysis was conducted by logistic regression. RESULTS: The prevalence of heavy alcohol consumption was 14.3% (29.2% among men and 3.7% among women). The following groups presented higher prevalences of heavy alcohol consumption after adjusted analysis: men, elderly people, blacks or mulattoes, heavy smokers, and people who present some kind of chronic disease. Men with minor psychiatric disorders showed higher prevalences of heavy alcohol consumption than other men. Among women, association between age and heavy alcohol consumption was inversely related. Furthermore, the study indicates that among hypertensive subjects, those with heavy alcohol consumption presented worse disease management. CONCLUSIONS: Heavy alcohol consumption is high and results in countless negative consequences for the individual's health and quality of life. Our results highlight the high prevalence of heavy alcohol consumption and indicate subsections of the whole population more susceptible to alcoholism. PMID- 15122387 TI - [Prevalence of asthma and asthma symptoms among 13 and 14-year-old schoolchildren, Brazil]. AB - OBJECTIVE: Before the use of the ISAAC (International Study of Asthma and Allergies in Childhood) questionnaire in epidemiologic surveys, little could be told about the comparative occurrence of asthma in the world due to differences in employed methods. In Brazil, the ISAAC questionnaire has been used in some urban regions. In this study it was applied in both, urban and rural areas, in order to estimate the prevalence of asthma among schoolchildren living in Montes Claros, Brazil. METHODS: A cross-sectional study was carried out using the ISAAC written questionnaire with four questions added on exposures of interest. The questionnaire was self-applied in 3,770 randomly selected schoolchildren aged 13 and 14 years. RESULTS: The prevalence of "wheezing in the last year" was 15.8%, and "asthma or bronchitis ever" was 23.8% with no statistically significant difference between boys and girls. There were significant differences between girls and boys regarding "wheezing ever" (37.8% and 33.6%), "sleeping disturbed by wheezing" (13.7% and 9.5%), and "nocturnal dry coughing without respiratory infection" (36.6% and 28.7%), respectively. "Wheezing in the last year" was found to be positively associated with "pet contact" (OR=1.27; 95% CI: 1.03-1.56), and "family history of asthma" (OR=1.79; 95% CI: 1.50-2.14), and negatively associated with "rural school" (OR=0.63; 95% CI: 0.44-0.91). But no association was found with sex, age, private/public school, and passive smoking. CONCLUSIONS: The prevalence of asthma in Montes Claros was high and some symptoms were seen mainly among girls. The occurrence of "wheezing in the last year" showed to be associated with family history, contact with pets and urban schools. PMID- 15122388 TI - [Evaluation of the program for prevention and health promotion in phenylketonuria patients in Brazil]. AB - OBJECTIVE: To evaluate the results of systematizing preventive and health promotion actions among phenylketonuria (PKU) patients. METHODS: Results of phenylketonuria patients attended in the Prenatal PKU Screening Program in the State of Parana, Brazil, from 1996 to 2001, were evaluated. Socioeconomic data were investigated and the gross motor function measure was applied to determine the motor score among 32 PKU infants with early diagnosis and treatment. Pearson's correlation coefficient was adopted to investigate the relationship between the target variable (motor score) and other quantitative variables (mean post-treatment serum phenylalanine level, parents' educational level, infant's age at the start of treatment, and family income). RESULTS: Among all the children evaluated, 93.7% showed development within normal limits as reported in the literature. Treatment was initiated in the first month of life in 71.9% of the PKU cases. Socioeconomic data showed 39.5% of parents having completed the fourth grade of primary school or less. There was a significant correlation between infant's motor score and parents' educational level (N=32), as well as between motor score and early treatment (N=27). CONCLUSIONS: The results highlighted the program's effectiveness. The correlation between parents' low educational level and lower motor score emphasizes the importance of support for parents in their use of diet therapy. The association between motor score and early initiation of treatment confirms the need for immediate admission into the program. The paucity of other evaluation studies in the literature makes generalization of the results difficult. PMID- 15122389 TI - [Methodology to evaluation the habitual physical activity in men aged 50 years or more]. AB - OBJECTIVE: To describe the methodology for evaluating habitual physical activity that was adopted for a survey among the male population utilizing a questionnaire validated earlier. METHODS: The Baecke questionnaire on habitual physical activity was translated into Portuguese and then back-translated into English by two anglophone teachers. The final version of the questionnaire was applied in a cross-sectional epidemiological study done on 326 men aged 50 years or over. The internal consistency among the questions was evaluated using the Cronbach a statistic. The Spearman correlation coefficients between the habitual physical activity scores were calculated. Partial correlation coefficients with adjustments for age, body mass index and schooling were also calculated. RESULTS: There was satisfactory internal consistency in relation to the magnitudes of occupational physical activity and leisure-time physical exercises. Significant correlation was obtained between all the physical activity scores and the total habitual physical activity score, independent of age, body mass index and schooling. CONCLUSIONS: The Baecke questionnaire was found to be a practical instrument for assessing habitual physical activity that is quickly applied and easily understood, and it is recommended for epidemiological studies in Brazil. PMID- 15122390 TI - [Estimate of the validity of a new method for the isolation of rabies virus]. AB - OBJECTIVES: No population-based studies have been conducted to show the potential for the use of virological diagnosis of the rabies virus. The objective of the present study was to estimate accuracy parameters for the isolation of the rabies virus in McCoy cells as an alternative method and to compare this with the use of murine neuroblastoma (N2A) cells, which is considered to be a reference method. METHODS: An evaluation was performed on 120 bats collected at random in the Atlantic Forest of the State of Sao Paulo. The immunofluorescence reaction was utilized for the detection of the rabies virus isolated from the brain of these bats and the presence of the virus was tested in the two cell culture systems. Two data sets were constructed with the results and the analysis was performed using the computer methods for diagnosis tests (CMDT) software by means of the two-graph receiver operating characteristic (TG-ROC) technique to determine sensitivity and specificity parameters, as well as other indicators such as efficacy, positive predictive value, negative predictive value and likelihood ratio. RESULTS: N2A cells presented 90% sensitivity and specificity, while McCoy cells presented 95% sensitivity and specificity. These values were based on cut off points optimized for each cell type. CONCLUSION: The study showed that McCoy cells allowed the obtaining of accuracy estimates that were better than for N2A. The McCoy cell method is therefore an effective method for the isolation of the rabies virus. PMID- 15122391 TI - [Parasitoids of Chrysomya megacephala (Fabricius) collected in Itumbiara, Goias, Brazil]. AB - This study determined the species of parasitoids associated with Chrysomya megacephala, collected on bovine kidney baits, in Itumbiara, State of Goias, Brazil. The pupae were obtained by flotation. They were individually placed in gelatin capsules until the emergence of the adult flies or their parasitoids. The overall prevalence of parasitism was 18.6%. Brachymeria podagrica, Nasonia vitripennis and Pachycrepoideus vindemiae presented frequencies of 8.6%, 8.6% and 1.4%, respectively. This work reports for the first time the occurrence of Brachymeria podagrica in pupae of Chrysomya megacephala. PMID- 15122392 TI - [Antibacterial activity of essential oils on microorganisms isolated from urinary tract infection]. AB - The antibacterial activity of essential oils extracted from medicinal plants (Ocimum gratissimum, L., Cybopogum citratus (DC) Stapf., and Salvia officinalis, L.) was assessed on bacterial strains derived from 100 urine samples. Samples were taken from subjects diagnosed with urinary tract infection living in the community. Microorganisms were plated on Muller Hinton agar. Plant extracts were applied using a Steers replicator and petri dishes were incubated at 37 degrees C for 24 hours. Salvia officinalis, L. showed enhanced inhibitory activity compared to the other two herbs, with 100% efficiency against Klebsiella and Enterobacter species, 96% against Escherichia coli, 83% against Proteus mirabilis, and 75% against Morganella morganii. PMID- 15122393 TI - [Clinical signs to diagnose anaemia: a late and inefficient action against iron deficiency]. PMID- 15122394 TI - [Epidemiology of American tegumentar leishmaniasis in the municipality of Ubatuba, North coastal area of the state of Sao Paulo, Brazil, 1993-2003]. PMID- 15122395 TI - [Clemente Ferreira Institute's review of adverse effects of tuberculosis drugs in children]. PMID- 15122396 TI - [Accreditation: promoting quality in health services]. PMID- 15122397 TI - [Importing medicines with quality assurance]. PMID- 15122398 TI - [Life at health care organizations]. PMID- 15122399 TI - [Undergraduate education at the University of Sao Paulo at Ribeirao Preto College of Nursing throughout the 50 years of its history (1953-2003)]. AB - This study aims to describe the course of undergraduate education developed at the University of Sao Paulo at Ribeirao Preto College of Nursing throughout the 50 years of its history (1953-2003), offering tools for reflection about the historical and social context, about current curricular guidelines and political and pedagogical challenges with a view to nursing formation. PMID- 15122400 TI - [Symptoms of stress in workers from five family health centers]. AB - This study aimed at investigating the occurrence of stress among workers at five family health centers of the University of Sao Paulo at Ribeirao Preto School of Medicine. The author used Lipp's Inventory of Stress Symptoms in Adults. The instrument aims to identify the symptoms presented by the individuals, evaluating their stress, the predominant kind of symptom and its phase of development. The author observed the presence of stress in 62% of the workers. Among them, 83% of the subjects were in the resistance phase and 17% in near-exhaustion. The psychological symptoms predominated in 48% of subjects, the physical in 39% and symptom equality was found in 13% of this group of workers. PMID- 15122401 TI - [Stressors in care at a thoracic surgery postoperative unit: nursing evaluation]. AB - The aim of this descriptive study was to determine what events are perceived as stressful to patients in the thoracic surgery postoperative unit, according to nursing evaluation. Data were collected at two hospitals, where the intensive care nursing team members completed the questionnaire. A 4-point Likert scale was used to evaluate 42 possible stressors. The sample consisted of 58 nursing professionals. The average duration of their professional activities at the intensive care unit was 5.9 years. The average stressfulness score of the items was 2.97 (from little stressful to stressful). The most important stressors were: having pain, having tubes in nose and/or mouth, being tied down by tubes and not being able to sleep. We concluded that, according to nursing evaluation, the main stressors for patients are associated with the surgical procedure. PMID- 15122402 TI - [Quality of life in nursing teaching]. AB - The research deals with the quality of life at work - QLW of nursing teachers. The main goal is to point the meanings and the extenuating and enabling QLW processes, generating the health-sickness profile. The theoretical background was constituted by social determination and by content analysis of the interviews. QLW mainly means working conditions, professional identification and interpersonal relationship. In the health-sickness profile, the extenuating factors exceed the enabling factors PMID- 15122403 TI - [Contamination risks caused by occupational accidents with cutting and piercing material among nursing workers]. AB - This study aimed to identify, among nursing workers from four hospitals in the region of Ribeirao Preto-SP, Brazil, victims of occupational accidents with cutting and piercing material, who were sent for evaluation at a service specialized in treating infectious diseases, individuals who were contaminated and the conduct adopted as a result of the accidents. This is a descriptive field research. The sample consisted of 30 subjects and data were collected by consulting the workers' medical files. The results showed that none of the workers had been contaminated by HBV, HCV or HIV. However, it was observed that only 23.33% of them had kept all the scheduled appointments in order to verify a possible serum conversion. Concerning the conducts adopted as a result of the accident, the use of chemoprophylaxis was recommended in 76.67% of the cases, serological tests in 100% and immunization against Hepatitis in 9.99%. Due to the high occurrence of percutaneous accidents, according to official estimates from other countries, it was concluded that more attention must be given to the prevention of these accidents, as well as to the strict follow-up of workers after occupational exposure. PMID- 15122405 TI - [Considerations on the process construction of nurse's identity in the daily of work]. AB - This qualitative study is part of the health and society area, work process and health knowledge sub-area. As a workforce study - the human element of work - it aimed to understand the nurse identity construction process on the basis of everyday work experience. Berger and Luckmann and Agnes Heller were the primary reference framework. This study was carried out at a public hospital in Cuiaba - Mato Grosso, Brazil. The nurse essentially revealed to be a being in construction in time, space and daily relations. To be a nurse means to have perceptions and expressions that can be materialized to a greater or lesser extent. The way of being a nurse indicates his/her material, sensitive and expressive forms, molded according to time, space and relations and representing a conjunction of the being and his/her perceptions/expressions. PMID- 15122404 TI - [Work ability among nursing workers at the emergency service of a university hospital]. AB - In a couple of years, the demographic and epidemiologic transition observed in Brazil will put this country in a worldwide position of prominence in relation to the aged population. Furthermore, the need for economic adaptation and consequent changes in Brazilian social security will result in workers staying in the labor market for a larger period of time, thus influencing their functional ability. This study aimed to evaluate work ability among nursing workers at the Emergency Service of a University Hospital. Study participants were fifty-four workers (40 women and 14 men), whose ages ranged from 23 to 53 years (average age 37.3 years). We used the Work Ability Index (WAI), an instrument developed in Finland, which presented an average value of 42.0. The most mentioned diseases with medical diagnosis were musculoskeletal, cardiovascular, respiratory and neurological. We consider that health promotion at work is one of the fundamental aspects for maintaining work ability. PMID- 15122406 TI - [Survey of types of repercussions as a result of performance evaluation in hospital nursing]. AB - This article is part of a study about performance evaluation in nursing and aims to identify and describe the measures adopted on the basis of the nursing staff performance evaluation that was carried out at municipal district hospitals in the Paraiba Valley Region - Sao Paulo, Brazil. Data were collected by means of a questionnaire, which was applied among the nurses in charge of the Nursing or Continuous Education Services. The obtained data allowed us to verify that, although 66.7% of the hospitals do not have a Continuous Education Service, 80.9% carry out performance evaluation and indicate the adoption of several measures according to the observed results. Educational measures prevailed, such as: orientation (97.0%), training (79.4%), updating/recycling (70.6%) and discussion of the result with the employee (88.2%). It was also verified that, in a significant percentage of the hospitals, the performance evaluation results do not only offer support for promotions (20.6%) and several incentives (58.8%), but also for employees' dismissal (58.8%). PMID- 15122407 TI - [Educational guideline for the maternal orientation concerning the care with preterm infants]. AB - This work aimed at describing the development of educational and instructional material for maternal training, so as to prepare the mother for the preterm infants' discharge from hospital, by means of the participatory methodology. The pedagogical model used was that of education for critical consciousness, based on Paulo Freire. Study participants were two nurses, two nursing auxiliaries and four mothers of preterm babies, which were hospitalized at the Intermediate Care Unit of a university hospital in Ribeirao Preto-SP, Brazil. The participants indicated the subjects of interest for the teaching-learning process, which were grouped into the categories: daily care, feeding, hygiene, special care and family relationship. We decided to develop an educational folder with figures, which could be taken home. This educational and instructional material was produced by the researchers on the basis of literature, their professional experience and on technical and scientific advice from other professionals. The final version of the folder was validated by the participants and now constitutes a creative instrument that can be of help in health education activities oriented towards these clients. According to the participants, the educational material directed the guidelines and helped the mothers to memorize the content that had to be learned. PMID- 15122408 TI - [The mothers' participation in the child malnutrition program]. AB - This study aims to show mothers' and health professionals' conceptions and perceptions related to their participation in the Child Malnutrition Program. Thus, we particularly explore the mothers' participation, as a means of making them aware of the individual and social reality and their decisions about it. Our analysis of the maternal participation process is guided by Imogene King's conceptual framework. This study was conducted in Crato, Ceara State, Brazil. Study participants were mothers and professionals. We used semi-structured interviews and adopted Bardin's Content Analysis for organizing and analyzing the collected data. This study did not find evidence of strategies for the conscious participation of social actors in the child malnutrition program. PMID- 15122409 TI - [Perroca's patient classification instrument: construct validity analysis]. AB - This paper starts from a research on the reliability and validity assessment of Perroca's patient classification instrument. Its purpose is to analyze the construct validity of this instrument, as well as to check its use in nursing management practice. Study participants were a hundred forty-one patients, allocated in Intensive Care and Hospitalization Units of a school hospital in the interior of Sao Paulo State, Brazil. Several statistical techniques were used to show the construct validity of the instrument. Research results revealed the importance of all critical indicators, each of which has its own role in determining the patient's complexity degree in relation to nursing care. Therefore, the study showed that the instrument shows evidence of validity. Hence, it is useful for nursing management practice, as a guide to the patient's care needs as well as to the nursing team's workload. PMID- 15122410 TI - [Logical-mathematical "ND" software development]. AB - In this article, we propose a mathematical analysis model for the generation of a proportional value of defining characteristics in nursing diagnoses. The study results are based on the logical-mathematical development of software specifically built for the generation of diagnosis hypotheses. The final calculation creates the sum of all defining characteristics found in each diagnosis, comparing it with the value of the V variable of the diagnosis, according to a mathematical rule that was set. It is concluded that the software needs to go through clinical validation in order to evaluate the pertinence of the rule proposed here. PMID- 15122411 TI - [Being a nursing teacher in an informatized world]. AB - The social phenomenology approach allowed us to understand that informatics projects are based on the biographic situation and the educational experiences of each teacher and their adoption happens voluntarily, in a reflection process on the favorable and unfavorable alternatives implied in the social context. Thus, we conjecture about the construction of teachers' technological and pedagogical competences and pro-active institutional policies that adhere to the contemporary society, seeking to integrate the new technologies with the needs of the profession and the human dimension of nursing. PMID- 15122412 TI - [Model for introducing or revitalizing the final monograph]. AB - The requirement set by the Curricular Guidelines for a Course Conclusion Work in the nursing area is an innovation for the majority of courses in Brazil. Only a few courses have introduced this type of study as a result of their forward looking vision. This requirement has demanded an effort from the universities, to ensure that these studies do not only represent an academic exercise, but also an institutional quality indicator and a possible contribution to the solution of social problems. Our proposed model includes: defining lines of research, gathering researchers per by area of interest, organizing research groups and centers, defining the preferred types of studies, planning operational agendas, carrying out a follow-up on their introduction and encouraging their publication. PMID- 15122413 TI - [Living the burden in becoming a family caregiver for a cerebrovascular accident survivor: knowledge analysis]. AB - This bibliographical research aims to carry out a thematic analysis of knowledge production in periodicals, about the burden placed on family caregivers of Cerebrovascular Accident (CVA) survivors. The analysis gathered articles found in the 80 and 90, through the Medline, Lilacs and Cinahl databases. The content analysis was the methodological background that enabled us to organize the whole knowledge into a body of categories and subcategories, called: Category 1 - The sequels of the CVA creating burden, Category 2 - Aspects generating burden, covering the following subcategories: social isolation, marital changes and dissatisfactions, family caregivers' financial difficulties and deficits in their physical health and self-care, Category 3 - Other analyses about burden in CVA survivors family caregivers. PMID- 15122414 TI - [Basic research in nursing]. AB - The integration between basic science and research in the field of nursing is an emerging subject and expanding in many countries. However, in Brazil, researchers have shown little interest in exploring this line of research, in spite of the numerous benefits it may bring to nursing care practice. In this article, by means of a bibliographic review, we will focus on the importance of basic research and take into consideration our own experiences and expectations as nursing researchers, emphasizing the possibility of integrating fundamental and experimental science with nursing research and practice. PMID- 15122415 TI - [Concept dimension for nursing and Deleuze]. AB - The study aimed at identifying and analyzing the meaning of concept according to Deleuze, in Philosophy, and Rodgers, in Nursing. Based on an extract of reality, Deleuze creates the concept and calls it "plan of immanence". For Rodgers, in nursing, the concept emerges at the actual moment one wishes to construct the conceptual bases for Nursing. The authors demonstrate that, although Deleuze and Rodgers come from different areas, the meaning they attribute to concept shares the happening of things and beings, which reflect the experienced reality. For Rodgers, the concept is characterized by its essential elements while, for Deleuze, by what he called components, that is, by essence. PMID- 15122416 TI - [The health labor market in the South-East of Brazil: the insertion of the nursing team]. AB - The study points out the main characteristics of the nursing labor market in the Brazilian South-East, using Medical-Sanitary Assistance-AMS research data as a reference base, which were recently published by the Brazilian Institute of National Statistics and Geography-IBGE. With 272398 jobs in this region, the nursing team mainly works in the public health system and hospital institutions. In coherence with the Brazilian health system decentralization policies, the municipalization of nursing jobs and signs of labor market flexibility were observed in 1999. In view of this national and regional context, a research should be carried to obtain a quantitative and qualitative analysis of the current labor market for the nursing team. PMID- 15122417 TI - Surgery information reduces anxiety in the pre-operative period. AB - PURPOSE: Patients preparing to undergo surgery should not suffer needless anxiety. This study aimed to evaluate anxiety levels on the day before surgery as related to the information known by the patient regarding the diagnosis, surgical procedure, or anesthesia. METHOD: Patients reported their knowledge of diagnosis, surgery, and anesthesia. The Spielberger State-Trait Anxiety Inventory (STAI) was used to measure patient anxiety levels. RESULTS: One hundred and forty-nine patients were selected, and 82 females and 38 males were interviewed. Twenty-nine patients were excluded due to illiteracy. The state-anxiety levels were alike for males and females (36.10 +/- 11.94 vs. 37.61 +/- 8.76) (mean +/- SD). Trait anxiety levels were higher for women (42.55 +/- 10.39 vs. 38.08 +/- 12.25, P = 0.041). Patient education level did not influence the state-anxiety level but was inversely related to the trait-anxiety level. Knowledge of the diagnosis was clear for 91.7% of patients, of the surgery for 75.0%, and of anesthesia for 37.5%. Unfamiliarity with the surgical procedure raised state-anxiety levels (P = 0.021). A lower state-anxiety level was found among patients who did not know the diagnosis but knew about the surgery (P = 0.038). CONCLUSIONS: Increased knowledge of patients regarding the surgery they are about to undergo may reduce their state-anxiety levels. PMID- 15122418 TI - Adductor pollicis muscle: a new anthropometric parameter. AB - PURPOSE: To measure the thickness of adductor pollicis muscle in healthy adults. This measurement will be used as a nutritional anthropometric parameter in further studies. SUBJECTS AND METHOD: Four hundred and twenty-one healthy adults were studied, 209 men and 212 women, with ages ranging from 18 to 87 years, living in Rio de Janeiro. The adductor pollicis muscle was also studied in the human anatomy lab as well as in normal healthy volunteers using CAT scans and nuclear magnetic resonance imaging to ensure that only the adductor pollicis was included in measurement of muscle thickness with a Lange caliper. To standardize the measurement, the methodology was detailed, with subjects sitting with the dominant hand dangling over the homolateral thigh and the elbow bent at approximately a 90 degrees angle. The Lange caliper was applied at a pressure of 10 g/mm2, pinching the adductor pollicis muscle at the vertex of an imaginary angle between the thumb and the index finger. The average of 3 consecutive measurements was considered to be the muscle thickness. RESULTS: This study provides the first estimates of adductor pollicis thickness in normal healthy subjects as an anthropometric parameter. The normal values in the dominant hand for men were 12.5 +/- 2.8 mm (mean +/- SD), median 12 mm, and for women were 10.5 +/- 2.3 mm, median 10 mm. PMID- 15122419 TI - Relationship between the morphologic alterations of vocal cords from adult autopsies and the cause of death. AB - PURPOSE: The purpose of this study was to identify the possible alteration in the thickness of the epithelium basal membrane of the vocal cords and correlate it with the cause of death. METHOD: Larynxes collected from adult autopsies during the period of 1993 to 2001 were utilized. We used the hematoxylin-eosin and periodic acid-Schiff staining methods for the morphological and morphometric analysis. RESULTS: Sixty-six vocal cords were analysed; increased thickness was identified in 14 cases (21.2%), with equal proportions between the genders. Increased vocal-cord thickness was more frequent in patients of the white ethnicity (12 cases, 85.7%). Respiratory alterations were found in 10 (71.4%) of the cases with increased vocal-cord thickness. Of the patients that were maintained with mechanical ventilation before death, 7 (18.4%) had thickening of the basal membrane. Among the smokers, 9 (19.63%) had basal membrane thickening. CONCLUSION: No statistically significant differences were found between the cases in which the cause of death was related to respiratory diseases as compared to non-respiratory diseases and the thickening of the basal membrane of the vocal cords. However, new studies are needed in order to verify the etiopathogenesis of this thickening. PMID- 15122420 TI - Growth and puberty after treatment for acute lymphoblastic leukemia. AB - Over the last 20 years, after combining treatment of chemotherapy and radiotherapy, there has been an improvement in the survival rate of acute lymphoblastic leukemia patients, with a current cure rate of around 70%. Children with the disease have been enrolled into international treatment protocols designed to improve survival and minimize the serious irreversible late effects. Our oncology unit uses the international protocol: GBTLI LLA-85 and 90, with the drugs methotrexate, cytosine, arabinoside, dexamethasone, and radiotherapy. However, these treatments can cause gonadal damage and growth impairment. PATIENTS AND METHOD: The authors analyzed 20 children off therapy in order to determine the role of the various doses of radiotherapy regarding endocrinological alterations. They were divided into 3 groups according to central nervous system prophylaxis: Group A underwent chemotherapy, group B underwent chemotherapy plus radiotherapy (18 Gy), and group C underwent chemotherapy plus radiotherapy (24 Gy). Serum concentrations of LH, FSH, GH, and testosterone were determined. Imaging studies included bone age, pelvic ultrasound and scrotum, and skull magnetic resonance imaging. RESULTS: Nine of the patients who received radiotherapy had decreased pituitary volume. There was a significant difference in the response to GH and loss of predicted final stature (Bayley-Pinneau) between the 2 irradiated groups and the group that was not irradiated, but there was no difference regarding the radiation doses used (18 or 24 Gy). The final predicted height (Bayley-Pinneau) was significantly less (P = 0.0071) in both groups treated with radiotherapy. Two girls had precocious puberty, and 1 boy with delayed puberty presented calcification of the epididymis. CONCLUSION: Radiotherapy was been responsible for late side effects, especially related to growth and puberty. PMID- 15122421 TI - Anatomic study of the dorsal arterial system of the hand. AB - Historically, the dorsal arterial system of the hand received less attention than the palmar system. The studies concerning dorsal arterial anatomy present some controversies regarding the origin and presence of the dorsal metacarpal artery branches. Knowledge of the anatomy of dorsal metacarpal arteries is especially applied in the surgical planning for flaps taken from the dorsum of the hand. The purpose of this study is to analyze the arterial anatomy of the dorsum of the hand, compare our observations with those of previous studies from the literature, and therefore to define parameters for surgical planning for flaps supplied by the dorsal metacarpal arteries. METHOD: Twenty-six dissections were performed at the dorsum of the right hand of 26 cadavers by making a distal-based U-shaped incision. After catheterization of the radial artery at the wrist level, a plastic dye solution with low viscosity and quick solidification was injected to allow adequate exposure of even small vessels. The radial artery and its branches, the dorsal arterial arch, the dorsal metacarpal arteries, the distal and proximal communicating branches of the palmar system, and the distal cutaneous branches were carefully dissected and identified. RESULTS: The distal cutaneous branches originating from the dorsal metacarpal arteries were observed in all cases; these were located an average of 1.2 cm proximal from the metacarpophalangeal joint. The first dorsal metacarpal artery presented in 3 different patterns regarding its course: fascial, subfascial, and mixed. The branching pattern of the radial artery at the first intermetacarpal space was its division into 3 branches. We observed the presence of the dorsal arterial arch arising from the radial artery in 100% of the cases. The distance between the dorsal arterial arch and the branching point of the radial artery was an average of 2 cm. The first and second dorsal metacarpal arteries were visualized in all cases. The third and fourth dorsal metacarpal arteries were visualized in 96.2% and 92.3% of cases, respectively. There was proximal and distal communication between the dorsal arterial arch and the palmar system through the communicating branches contributing to the dorsal metacarpal artery formation. CONCLUSION: At the dorsum of the hand there is a rich arterial net that anastomoses with the palmar arterial system. This anatomical characteristic allows the utilization of the dorsal aspect of the hand as potential donor site for cutaneous flaps. PMID- 15122422 TI - Pulmonary artery catheter complications: report on a case of a knot accident and literature review. AB - A particular event concerning a Swan-Ganz catheter complication is reported. A 41 year-old woman was admitted at the emergency room of our hospital with massive gastrointestinal bleeding. A total gastrectomy was performed. During the postoperative period in the intensive care unit, the patient maintained hemodynamic instability. Invasive hemodynamic monitoring with a pulmonary artery catheter was then indicated. During the maneuvers to insert the catheter, a true knot formation was identified at the level of the superior vena cava. Several maneuvers by radiological endovascular invasive techniques allowed removal of the catheter. The authors describe the details of this procedure and provide comments regarding the various techniques that were employed in overcoming this event. A comprehensive review of evidence regarding the benefits and risks of pulmonary artery catheterization was performed. The consensus statement regarding the indications, utilization, and management of the pulmonary artery catheterization that were issued by a consensus conference held in 1996 are also discussed in detail. PMID- 15122423 TI - Titanium pigment in tissues of drug addicts: report of 5 necropsied cases. AB - The aim of this report is to describe the anatomic-pathologic findings from necropsies of 5 drug addicts with titanium pigment in several organs after chronic intravenous injection of crushed propoxyphene hydrochloride tablets. Samples from liver, spleen, lungs, lymph nodes, and bone marrow were obtained, and after being grossly studied, they were submitted to evaluation using common light and polarized microscopy. In all 5 cases, a pigment with characteristics of titanium dioxide was found within tissue samples of the organs studied. Our findings suggest that research concerning titanium pigment within body tissues should be enhanced, considering the potential contribution of this morphologic data to forensic pathology. PMID- 15122424 TI - Hydronephrosis in Schinzel-Giedion syndrome: an important clue for the diagnosis. AB - Schinzel-Giedion syndrome is a rare autosomal recessive disorder characterized by coarse facies, midface retraction, hypertrichosis, multiple skeletal anomalies, and cardiac and renal malformations. Craniofacial abnormalities of this syndrome sometimes resemble a storage or metabolic disease. The pathogenesis of the disease remains unknown. The objective of this report was to emphasize the importance of congenital bilateral hydronephrosis for the diagnosis of Schinzel Giedion syndrome. We describe the first Brazilian case of a newborn with typical facies, generalized hypertrichosis, cardiac and skeletal anomalies, and bilateral hydronephrosis detected during pregnancy and confirmed later by abdominal ultrasonography. Chromosomal constitution was normal. Of the 35 cases already reported in the literature, 31 presented hydronephrosis, which is considered an important clue in diagnosis. If Schinzel-Giedion syndrome were indexed as a cause of congenital hydronephrosis, its identification would be greatly facilitated, since the majority of the other findings in Schinzel-Giedion syndrome are nonspecific and common to many genetic syndromes. PMID- 15122425 TI - Lateralization of epileptiform discharges in patients with epilepsy and precocious destructive brain insults. AB - Unilateral destructive brain lesions of early development can result in compensatory thickening of the ipsilateral cranial vault. The aim of this study was to determine the frequency of these bone changes among patients with epilepsy and precocious destructive lesions, and whether a relationship exists between these changes and epileptiform discharges lateralization. Fifty-one patients had their ictal / interictal scalp EEG and skull thickness symmetry on MRI analyzed. Patients were divided into three main groups according to the topographic distribution of the lesion on the MRI: hemispheric (H) (n=9); main arterial territory (AT) (n=25); arterial borderzone (Bdz) (n=17). The EEG background activity was abnormal in 26 patients and were more frequent among patients of group H (p= 0.044). Thickening of the skull was more frequent among patients of group H (p= 0.004). Five patients (9.8%) showed discordant lateralization between epileptiform discharges and structural lesion (four of them with an abnormal background, and only two of them with skull changes). In one of these patients, ictal SPECT provided strong evidence for scalp EEG false lateralization. The findings suggest that compensatory skull thickening in patients with precocious destructive brain insults are more frequent among patients with unilateral and large lesions. However, EEG lateralization discordance among these patients seems to be more related to EEG background abnormalities and extent of cerebral damage than to skull changes. PMID- 15122426 TI - The role of mirror focus in the surgical outcome of patients with indolent temporal lobe tumors. AB - PURPOSE: To review the clinical and neurophysiological data of 21 patients with epilepsy due to temporal lobe tumors and who had undergone evaluation and surgery at the Hospital das Clinicas da Universidade de Sao Paulo. The aim of this study was to investigate whether the occurrence of a mirror focus was influenced either by certain clinical factors or if the surgical outcome was influenced by the presence of a mirror focus. METHOD: We included these 21 patients who had undergone at least one interictal electroencephalogram in the pre- and post surgical periods. They had had a minimum follow-up of one year. RESULTS: Eight patients had mirror focus (Group 1) and 13 did not (Group 2). The mean age at seizure onset, duration of epilepsy disorder and total number of seizures did not vary statistically between the two groups of patients. Generalized tonic-clonic seizures occurred more frequently in the mirror focus group. All, but one patient, with a mirror focus were seizure free at follow- up. The mirror focus disappeared in all eight patients in the post-surgical electroencephalogram. In this group, the patient who was not seizure - free had a seizure recorded in his post-surgical electroencephalogram with seizure onset ipsilateral to the resected tumor. The patients who were not seizure-free had either been submitted to an incomplete resection of the tumor or showed evidence of associated cortical dysplasia. CONCLUSION: The occurrence of mirror focus is not a contraindication to surgery even when interictal epileptiform activity predominates contralaterally to the tumor and neither when seizures appear to arise from the mirror focus on scalp EEG. Good surgical outcome is expected despite EEG findings that may conflict with tumor location. PMID- 15122427 TI - Value of extent of hippocampal resection in the surgical treatment of temporal lobe epilepsy. AB - OBJECTIVE: Unilateral hippocampal atrophy is indicator of good surgical prognosis in patients with temporal lobe epilepsy (TLE). Some patients however do not become seizure free after surgery. We assessed if the extent of hippocampal and amygdala resection is associated with outcome. METHODS: Thirty patients with TLE with unilateral or clearly asymmetric hippocampal atrophy who underwent surgical treatment were evaluated concerning preoperative clinical variables and interictal EEG abnormalities. Amygdala and hippocampal resection was evaluated by post-operative MRI. We compared seizure free versus non-seizure free patients, and patients with good outcome (Engel's classes I and II) versus patients with poor outcome. RESULTS: There was significant association between the extent of hippocampal resection and the outcome. Pre-operative variables and interictal EEG abnormalities did not show relationship with outcome as documented in previous studies. CONCLUSION: The extent of hippocampal resection is associated with outcome. Incomplete resection of atrophic hippocampus may explain most surgical failures in patients with TLE due to unilateral hippocampal sclerosis. PMID- 15122428 TI - Experimental therapy of epilepsy with transcranial magnetic stimulation: lack of additional benefit with prolonged treatment. AB - OBJECTIVE: To investigate the effect of three months of low-frequency repetitive transcranial magnetic stimulation (rTMS) treatment in intractable epilepsy. METHODS: Five patients (four males, one female; ages 6 to 50 years), were enrolled in the study; their epilepsy could not be controlled by medical treatment and surgery was not indicated. rTMS was performed twice a week for three months; patients kept records of seizure frequency for an equal period of time before, during, and after rTMS sessions. rTMS was delivered to the vertex with a round coil, at an intensity 5% below motor threshold. During rTMS sessions, 100 stimuli (five series of 20 stimuli, with one-minute intervals between series) were delivered at a frequency of 0.3 Hz. RESULTS: Mean daily number of seizures (MDNS) decreased in three patients and increased in two during rTMS--one of these was treated for only one month; the best result was achieved in a patient with focal cortical dysplasia (reduction of 43.09% in MDNS). In the whole patient group, there was a significant (p<0.01) decrease in MDNS of 22.8%. CONCLUSION: Although prolonged rTMS treatment is safe and moderately decreases MDNS in a group of patients with intractable epilepsy, individual patient responses were mostly subtle and clinical relevance of this method is probably low. Our data suggest, however, that patients with focal cortical lesions may indeed benefit from this novel treatment. Further studies should concentrate on that patient subgroup. PMID- 15122430 TI - Epidemiologic features of Guillain-Barre syndrome in Sao Paulo, Brazil. AB - INTRODUCTION: There are few epidemiologic studies concerning Guillain-Barre syndrome (GBS). Due to difficulties with definition and lack of a standard diagnostic test of reference, GBS is not easy to study epidemiologically. We evaluate some epidemiological features of GBS in a sample of cases treated at a tertiary hospital in Sao Paulo, Brazil. METHOD: We retrospectively reviewed all cases of GBS with hospitalization in Santa Marcelina hospital, over the period of January 1995 through December 2002. RESULTS: Ninety-five cases were included in this study. Fifty-five were men and forty women, with a proportion of 1.4 men to 1 woman. The age ranged from 1 to 83 years with a mean age at onset of 34 years. GBS was less frequently observed below 15 years (18.9%) and above 60 years (16.9%). The highest frequency was observed in patients aged 15 to 60 years old (66.2%). The annual incidence rate was 0.6 cases/100,000 people. There was a highest frequency of cases during the months of September through March (62.1%). CONCLUSION: Our data differs from that of other epidemiological studies in that we did not observe a bimodal distribution in age and found a seasonal pattern in hotter months. PMID- 15122429 TI - Nonthyroidal illness syndrome in patients with subarachnoid hemorrhage due to intracranial aneurysm. AB - We have previously reported that subarachnoid hemorrhage due to ruptured intracranial aneurysm (SH) is associated with changes in the hormonal profile in the first 24 hours after the event. We proposed that the hormonal changes observed are due to the intense stress to which the patients are exposed. However, the thyroidal hormonal profile is indicative of the presence of a nonthyroidal illness syndrome (NTIS). In this paper, we examined whether the change in the thyroid hormone profile is compatible with a NTIS. Two groups of patients were included in the study: A) 30 patients with SH (21 females and 9 males; 41.7+/-11.4 years) and B) a control group including 25 patients with benign diseases of the spine (BDS) (lumbar disc hernia or stable spinal trauma) (8 females and 17 males; 41.3+/-14.2 years). In a subgroup of eight patients of each group serum triiodothyronine (T3) and reverse T3 levels were measured. The blood samples were obtained between 8:00 and 9:00 AM. The following results were obtained: The SH group had smaller serum T3 and free T4 levels than the BDS group (p<0.05): T3 (ng/mL): SH = 58.7+/-1.1 and BDS = 74.5+/-13.9; free T4 (ng/dL): SH = 0.9+/-0.2 and BDS = 1.1+/-0.3. There was no significant difference in the serum levels of total thyroxine (T4) and thyroid-stimulating hormone (TSH) between the two groups: T4 ( microg/dL): SH = 6.9+/-1.1 and BDS = 7.4+/-2.1; TSH ( microUI/mL): SH = 1.5+/-0.8 and BDS = 1.8+/-1,0. In the sample of eight patients of each group we had the following results: T3 (ng/mL): SH = 66.8+/-3.8 and BDS = 77.2+/-1.1 (p <0.05); reverse T3 (ng/dL): SH = 32.8+/-8 and BDS = 24.7+/-2.2 (NS); T3/ reverse T3 ratio: SH = 2.6+/-0.3 and BDS = 3.3+/-0.4 (NS). Thyreoglobulin and microsomal antibodies were not detectable, except in one patient in the SH group. In conclusion, the SH patients present serum levels of T3 and free T4 significantly lower than that of BDS patients; the thyroidal hormone profile suggests that SH patients have developed the nonthyroidal illness syndrome. PMID- 15122431 TI - Cerebral infarct in children aged zero to fifteen years. AB - Cerebral infarcts in children present peculiar characteristics either due to their diversity of causes or due to the unknown nature of the causes. The etiologies of cerebral infarct were reviewed in children from zero to 15 years old, attended at a tertiary hospital, in Ribeirao Preto (Brazil), from 1990 to 1997, adopting the modified Trial of ORG 10172 in Acute Stroke Treatment (TOAST) criteria of classification; 1-Atherosclerosis in large arteries; 2-Cardioembolic; 3-Occlusion of small vessels; 4-Other etiologies; 5-Undetermined cause. Thirty nine children were included, 18 males and 21 females, aged 2 months to 15 years, mean age 5.67. The largest group, N=22 (56.4%), included children with "other etiologies", 7 of them aged under two years. The most common etiology was dehydration and septic shock leading to brain hypoperfusion and watershed infarcts. Nine (23%) children had "Undetermined etiology", 7 (17.9%) cardioembolic subtype and none had atherosclerosis. Laboratory improvement is needed for the large number of patients without a defined cause, and the high proportion of children with dehydration in the group with a determined cause emphasizes the need for preventive health actions among infants and children. PMID- 15122432 TI - Quantitative electroencephalography (qEEG) to discriminate primary degenerative dementia from major depressive disorder (depression). AB - Electroencephalography (EEG) can be a valuable technique to assess electrophysiological changes related to dementia. In patients suspected of having dementia, the EEG is often quite informative. The sensitivity of the EEG to detect correlates of psychiatric disorders has been enhanced by means of quantitative methods of analysis (quantitative EEG). Quantitative features are extracted from, at least, 2 minutes of artifact-free, eyes closed, resting EEG, log-transformed to obtain Gaussianity, age-regressed, and Z-transformed relative to population norms (Neurometrics database). Using a subset of quantitative EEG (qEEG) features, forward stepwise discriminant analyses are used to construct classifier functions. Along this vein, the main objective of this experiment is to distinguish profiles of qEEG, which differentiate depressive from demented patients (n = 125). The results showed that demented patients present deviations above the control group in variables associated to slow rhythms: Normed Monopolar Relative Power Theta for Cz and Normed Bipolar Relative Power Theta for Head. On the other hand, the deviation below the control group occurs with the variable associated to alpha rhythm: Normed Monopolar Relative Power Alpha for P3, in dementia. Using this method, the present investigation demonstrated high discriminant accuracy in separating Primary Degenerative Dementia from Major Depressive Disorder (Depression). PMID- 15122434 TI - Performance of university students on random number generation at different rates to evaluate executive functions. AB - OBJECTIVE: To evaluate the performance of adult young subjects in a Random Number Generation (RNG) task by controlling the response speed (RS). METHOD: Sixty-nine university students of both sexes took part in the experiment (25.05 +/- 6.71 year-old). Participants were alloted into 3 groups which differed in RS rates to generate numbers: 1, 2 and 4 seconds to generate each number. A digital metronomer was used to control RS. Participants were asked to generate 100 numbers. The responses were measured through Evans's RNG Index. RESULTS: There were statistically significant differences among the groups [F (3, 68) = 7.120; p <.05]. Differences were localized between 1 and 2 seconds (p = 0.004) and between 1 and 4 seconds (p = 0.006). No differences were observed between 2 and 4 seconds (p = 0.985). CONCLUSION: The present results suggest that the response speed in production of random numbers influences the performance of the Random Numbers Generation task. PMID- 15122433 TI - Contribution to the evaluation of language disturbances in subcortical lesions: a pilot study. AB - Subcortical structures are in a strategic functional position within the cognitive networks and their lesion can interfere with a great number of functions. In this study, we describe fourteen subjects with exclusively subcortical vascular lesions (eight in the basal ganglia and six in the thalamus) and the interrelation between their language alterations and other cognitive abilities, as attention, memory and frontal executive functions. All patients were evaluated through the following batteries: Boston Diagnostic Aphasia Examination, Boston Naming Test, Token Test, Benton Visual Retention Test, Trail Making, Wisconsin Card Sorting Test and a frontal scripts task. All patients underwent MRI and twelve underwent SPECT. Results show that these patients present impairment in several cognitive domains, especially attention and executive functions. These alterations affect language abilities, and this fact must be considered in the rehabilitation efforts. PMID- 15122435 TI - [Functional magnetic resonance imaging in the determination of dominant language cerebral area]. AB - Functional magnetic resonance imaging (fMRI) is a technique for detecting minimal changes in brain perfusion and oxygenation secondary to neuronal activation. Its application in the pre-surgical evaluation of epileptic patients with temporal mesial sclerosis is currently being under investigation in several centers. This study aims to describe an activation paradigm for the evaluation of language and memory functions, as an alternative to the worldwide used Wada test, which is an invasive procedure. In order to propose a paradigm adapted to the Portuguese language, we report our experience in determining the dominant cerebral area for language through fMRI with a verbal fluency task. The results of the fMRI from 19 patients studied in Curitiba in a period of approximately two years were studied. Sixteen of them presented with left hemispheric cerebral language dominance. In five patients, results from fMRI and Wada test could be compared and agreed in localization. Our results reinforce the view that fMRI may become an essential tool for medical practice, perhaps for the determination of eloquent areas in the evaluation of candidates for epilepsy surgery. PMID- 15122436 TI - [Intracranial blood flow velocities evaluated by color Doppler (duplex) in preterm infants]. AB - In order to ascertain the blood flow velocities in the intracranial arteries we evaluated 73 preterm neonates during a period ranging from June 1994 to March 1999. These preterm infants were divided in two separate groups, 18 healthy and 55 with intracranial hemorrhage. They were subjected to sequential measurements of blood flow velocities in the intracranial arteries. The gestational age of the whole group varied from 28 to 36 weeks and birth weights between 720 and 2530 g. The diagnosis of the intracerebral hemorrhages in these preterm neonates were done using high resolution gray and color scale transfontanellar ultrasonography brain scans. The ultrasound evaluations were performed in the initial 3rd, 7th and 14th day of life. The 73 preterm infants were evaluated with sequential measurements of blood flow velocity in the intracranial arteries using the Doppler technique through the anterior fontanelle. Doppler evaluation of the cerebral vessels were performed on days 3, 7, 30 and 90 of life. These evaluations were performed in the six intracranial arteries, meaning: right and left anterior and middle cerebral arteries and right and left internal carotid arteries. Doppler recordings were made using Duplex Color-Doppler system, pulse echo probe of 3,5; 5,0 and 7,5 MHz. Measuring the blood flow velocity in the cerebral arteries we obtained a maximum systolic velocity and end diastolic velocity with a rate in meters per second (m/s) for each cardiac cycle. After obtaining these numerical values for these velocities we obtained the resistance index (RI) or Pourcelot index. In a progressive way as the resistance index (RI) values were being obtained in each stage of this study they were also being checked in the cerebral arteries of healthy preterm infants and infants with intracranial hemorrhages. We also analyzed in a comparative method the values of the resistive index between the two groups of preterm infants observing their behaviour. The results obtained when comparing the RI values in the various arteries during the different stages of the study permitted us to conclude that the RI values of healthy pre-term infants were always larger than the RI values of pre-term infants with intracranial hemorrhage. We also conclude that the RI values in the healthy pre-term infants and in the pre-term infants with intracranial hemorrhages decreased progressively with the increasing age of neonates. PMID- 15122437 TI - [Cognitive disorders and dementias in the private practice of specialists: pilot study of a self-assessment survey]. AB - OBJECTIVES: To know about the features of cognitive disorders and dementias in the private practice of specialists and these doctor's skills on that area. METHOD: In this pilot study, self-assessment questionnaires were delivered to neurologists of Sao Paulo State and 196 (22.8%) were respondents. RESULTS: Many neurologists are involved, besides the private practice, with teaching (61.5%) and/or research (59.5%) activities. Most of them assessed as not good the training on cognitive disorders and dementias they had had during both the graduate (77.3%) and residence (63.1%) courses; nevertheless 60.8% self rated their knowledge on that subject as satisfactory and 83.0% declared their interest on it as at least equal to other areas. The most frequent cognitive complaints occurring as primary reason for appointment are memory loss (73.0%) and attention/concentration deficits (48.0%). Dementia of Alzheimer type (54.9%) and vascular dementia (23.0%) are the most frequent ones in the neurologist private practice. CONCLUSION: Cognitive disorders and dementias represent a significant proportion in the neurological private practice. Although they had not had a good training on the area of cognitive disorders and dementias, the respondent neurologists demonstrated great interest on it. PMID- 15122438 TI - [Analysis of methylphenidate's efficacy using the abbreviated version Conners' questionnaire in attention deficit hyperactivity disorder]. AB - The attention deficit/hyperactivity disorder (ADHD) is a complex diagnosis. Its installation is precocious, with chronic evolution that impacts on the subject's performance in several contexts. Three to 5% of the school aged children present this disorder. The Conners' questionnaire has been used as an instrument of epidemiological survey for ADHD children. OBJECTIVE: This study aimed to investigate if this instrument could be an useful tool for the analysis of the methylphenidate's treatment efficacy in ADHD's children. METHOD: Twenty-one male children were selected, with ADHD combined type, chronological ages ranging from seven to 10 years and 11 months and all were treated with methylphenidate. The Conners' questionnaire abbreviated version for parents and teachers was applied in two moments, one without medication and another between six to eight months after the beginning of the methylphenidate. RESULTS: Results pointed to a reduction in punctuation in Conners' questionnaire in all children with ADHD concomitant to clinical improvement. CONCLUSION: Conners' questionnaire revealed to be useful not only for diagnosis, but also as an instrument to evaluate the effectiveness of the treatment of ADHD. PMID- 15122439 TI - [Clinical response to long action propranolol in 40 patients diagnosed with essential tremor with no previous treatment: an open, non-controlled study]. AB - Essential tremor (ET) is the most common movement disorder and betablockers are still considered the first line of treatment. The aim of our study is to report the clinical response to long action propranolol (LAP) of 40 patients diagnosed with essential tremor with no previous treatment. METHOD: 40 patients with ET were evaluated with rating scales for severity of tremor and clinical classification of ET. All patients were evaluated at least twice, at enrollment and one month after starting treatment. RESULTS: thirty-six patients (90%) had type 2 ET and types 3 and 4 occurred in two patients each (10%). Familiar history was positive in 25 patients (62.5%). Mean age at first evaluation was 43.1 years and mean age at onset was 27.4 years. Of all patients, 33 (82.5%) had some degree of benefit and in 52.5 % this benefit was either good or excellent. CONCLUSION: LAP seems to be a good treatment option for ET in our series of 40 patients. PMID- 15122440 TI - [Topiramate in the preventive treatment of migraine: experience in a tertiary center]. AB - Frequent migraine attacks require prophylactic treatment. Anticonvulsants have been suggested due to the progressive knowledge that cortical hyperexcitability is involved in migraine pathophysiology. Topiramate is one of these drugs and its efficacy has been demonstrated in several studies. The aim of this study is to evaluate the adherence and response to topiramate in migraineurs under treatment in a tertiary center. During a 2-year period, all of the patients receiving topiramate for migraine were evaluated after 3 months. The parameters evaluated were adherence to treatment, frequency reduction of attacks >50% and adverse events. Among 175 patients included, 134 (76.6%) returned. Among the 134 patients evaluated, 82 (61.2%) revealed frequency reduction >50% and 105 (78.4%) patients presented weight loss (average 3.4Kg). The most frequent side effects were paresthesias (39.6%); emotional disturbances (including depression, irritability and anxiety) in 17,9%; thinking impairment (12.7%); memory disturbances (12.7%) and altered taste (11.9%). Despite methodological limitations we concluded that adherence to its use and efficacy occurred in most of the patients. In addition, the side effect profile was acceptable. Further controlled studies are necessary to confirm these observations. PMID- 15122442 TI - [Craniotomy without trichotomy: analysis of 640 cases]. AB - The hair shaving in preparation for neurosurgery is frequently used in most of neurosurgical centers to perform craniotomy. We question about its necessity after our retrospective analysis of 640 patients undergoing cranial procedures without previous hair shaving. We had the overall surgical wound infection rate of 1.09%, not higher than tricotomy in the review of the literature. In 7 cases with infection, 3 patients were undergoing to CSF shunts, 3 patients had head injury, and one had brain tumor. The technique for preparing skin and hair for cranial procedures, its advantages and disadvantages are described and discussed. PMID- 15122441 TI - [Patients with ischemic stroke are taken care quickly in Sao Paulo Hospital]. AB - We observed inpatients with the diagnosis of ischemic stroke hospitalized at least 24 hours. We investigated the reasons for early or late arrival to the Emergency Room, the time span from arrival to computed tomography (CT) scanning, factors associated to hospitalization time, and patients outcome. We concluded that people who are cared for by Sao Paulo Hospital now get to the hospital sooner than they did 3 years ago (47% arrived in 3 hours or less) and that the patient flow inside the hospital became much faster, once any stroke patient is considered now a medical emergency by the hospital staff. Infections were the main complications presented by our patients, which is leading us to optimize protocols to improve emergency care by all the staff. PMID- 15122443 TI - [Validation of the functional assessment of multiple sclerosis quality of life instrument in a Portuguese language]. AB - The objective of the present study was to translate, to do cultural equivalence and validation of the Functional Assessment of Multiple Sclerosis quality of life instrument (FAMS) to Portuguese. The database analyzed is longitudinal, with 143 patients with clinically defined multiple sclerosis (MS). The validity of the scale was established by correlating total FAMS scores and sub-scales scores to EDSS and EAN. The internal consistency reliability were similar to those of the original FAMS. All sub-scales correlated with each other (0.35 to 0.71). Total FAMS score correlated significantly with EDSS, and EAN. All sub-scales correlated to EDSS. Kendall's Tau coefficient, showed significant associations between basal and first visit. Responsiveness was investigated considering two groups: patients with EDSS < 3.5 and with EDSS > 3.5. There was significantly higher score in the group with light EDSS when compared to the group of patients with moderate EDSS. One can conclude, based on this retrospective study that FAMS instrument translated to Brazilian Portuguese can be considered reliable, responsive and content valid. PMID- 15122444 TI - [Neural conduction in hand nerves and the physiological factor of age]. AB - Many researchers have established the influence of physiological factors as age, for the parameters of the study of the motor and sensitive conduction. The objective of this study was to analyze the influence of the variable age in the study of the motor and sensitive nervous conduction of the median and ulnar nerves. The data were collected from 92 volunteers: 61 women and 31 men. Their age was from 13 to 74 years old, with a mean of 36.3 years. Most of them were employees at Santa Casa de Sao Paulo. It was observed that a reduction in the velocity of sensitive and motor nervous conduction takes place with the age. This reduction is associated with a reduction in the amplitude of the evoked potential. PMID- 15122445 TI - [Progressive muscular atrophy: clinical and laboratory study in eleven patients]. AB - Progressive muscular atrophy (PMA), an infrequent type of motor neuron disease (MND), is a predominantly lower motor neuron degeneration, causing muscle wasting and weakness with loss of weight and fasciculations. The diagnosis is based on rigid criteria, considering clinical aspects and eletroneuromyography findings. Blood tests and radiological investigation are necessary to look for other diagnosis mimicking PMA. We herein present 11 patients with PMA (5.9% of all our MND patients), 9 men and 2 women, which onset of symptoms occurred mainly under de age of 50, with a mean of 45.5 years. Cramp was the most frequent symptom preceding muscular weakness. Muscle pain, fatigue and fasciculations were also cited as starting symptoms. Asymmetric weakness of the arms was the most frequent pattern of onset of the disease. Bulbar muscular weakness developed in all patients during the course of the disease. Predisposing factors and distinctive clinical outcome was not observed in any of the patients. Ophthalmoparesis and sphincter dysfunction were seen in two patients who had a prolonged time in artificial respiratory assistance. Immunosuppressive therapy was ineffective in all patients. Progressive course was seen in all cases and the mean survival time was 44 months. PMID- 15122446 TI - [Choroid plexus tumours: epidemiologic comparative study of 24 cases]. AB - Tumours derived from choroid plexus are rare central nervous system neoplasms affecting mainly children. This study presents a series of 38 patients with neuroectodermal tumours. Twenty four of them had the histological and/or immunohistochemical diagnosis of choroid plexus carcinoma (CPC). Sixteen of these patients were male. The left lateral ventricle was affected in most cases. Main clinical features were: hydrocephalus, intracranial hypertension and convulsion. All the patients were treated by surgery. There were 2 deaths due to surgical complications. Nine patients had recurrence, dying in a intermediate time of 12.3 months and 7 patients remain alive. The other cases did not present available data. CPC is very prevalent in Curitiba when compared to other choroid plexus tumours and it might be related to some pathogenic agent. PMID- 15122448 TI - Acute hemorrhagic leukoencephalitis mimicking herpes simplex encephalitis: case report. AB - Acute hemorrhagic leukoencephalitis (AHLE) is a more severe form of acute disseminated encephalomyelitis (ADEM) characterized by a fulminant clinical course and the presence of hemorrhagic necrosis of the white matter. We report the case of a 57-year-old woman who developed delirium following a respiratory infection. Magnetic resonance imaging of the brain disclosed signal abnormalities in the frontal and temporal lobes, usually found in herpes simplex encephalitis (HSE). Gram stain, India ink and acid-fast bacilli staining were all negative in CSF as was a polymerase chain reaction (PCR) for herpes simplex virus. A diagnosis of AHLE was made and the patient was treated with i.v. methylprednisolone 1g/day for 5 days. Despite treatment, the patient developed several neurological sequelae compatible with the severity of her illness. PMID- 15122447 TI - [Ultrastructural study of the remyelinating process following local ethidium bromide injection in the brainstem of dexamethasone-immunosuppressed rats]. AB - Lymphocytes are present within ethidium-bromide (EB)-demyelinated lesions in the central nervous system (CNS) and the possibility of its participation in accidental immune-mediated responses to the detached myelin sheaths can not be ruled out. This study aimed to investigate the consequences of immunosuppression with dexamethasone in CNS repair after local EB injection. Adult Wistar rats received 10 microlitres of 0.1% EB solution into the cisterna pontis. Some were treated intraperitoneally with dexamethasone (3 mg/kg/day, group I, n=15) during the experimental period; others were not immunosuppressed (group II, n=15). Animals from both groups were perfused with 4% glutharaldehyde at 7,11,15,21 and 31 days following EB injection. Brainstem slices were collected and processed for transmission electron microscopy studies. Rats from group I showed greater amounts of myelin-derived membranes than non-immunosuppressed rats (group II), suggesting a delay in the macrophagic activity of removing myelin debris. Rare lymphocytes were found. Oligodendrocyte remyelinating activity also showed a delayed pattern, with clear predominance of naked axons. PMID- 15122449 TI - Guillain-Barre syndrome in the course of dengue: case report. AB - This case report describes the findings of a 45-year-old white woman from Brazil, who developed myalgia, fever and macular rash. She was diagnosed as having dengue, based on clinical manifestations and specific IgM titers. One week after the first symptoms of dengue, the patient developed muscle weakness, followed by tetraplegia with areflexia, and respiratory insufficiency. The electromyography had evidence of demyelinating neuropathy and the cerebrospinal fluid showed albuminocytologic dissociation. These neurologic findings were consistent with the diagnosis of Guillain-Barre syndrome. The patient was treated with immunoglobulin and metylprednisolone. Mechanical ventilation was started one week after hospital admission and maintained for four weeks. After six weeks of hospitalization the patient was discharged from the hospital on wheel chair, presenting mild muscle weakness and loss of patellar and ankle reflexes. When the patient was seen at the outpatient service three weeks after hospital discharge she was able to walk with help. This case report suggests a possible association between dengue and Guillain-Barre syndrome. PMID- 15122450 TI - Meningoradiculitis due to Cryptococcus neofermans in an immunocompetent patient. AB - Meningoradiculitis refers to combined involvement of meninges and nerve roots. The most frequent location is the lumbosacral region. Etiology is diverse, including inflammatory, infectious and neoplastic disorders. Meningoradiculitis is a rare form of involvement in cryptococcal infection. We describe a case of subacute lower limbs flaccid paresis diagnosed as lumbosacral meningoradiculitis in view of cerebrospinal fluid (CSF) inflammatory changes and typical enhancement on MRI of lumbar spine. Cryptococcus neoformans was isolated from CSF. Extensive screening yielded no immunodeficiencies. PMID- 15122451 TI - [Peripheral neuropathy and myositis in idiopathic hypereosinophilic syndrome: case report]. AB - We describe a case of idiopathic hypereosinophilic syndrome manifested by an axonal sensory-motor polyneuropathy (ASMP) and signals of myositis. A 20 year old woman began with progressive gait impairment with drops and presented with subacute lower limb edema associated with paresthesis and cramps. She showed hypotonia in the lower limbs, absence of knee and ankle jerks, steppage gait, and decreased sensation on both legs. Examinations of stools were negative. Blood examination showed 7700 leukocytes with 24% (1848/mm(3)) eosinophils. Electrodiagnostic studies showed axonal lesion in sensory and motor nerves. Muscle biopsy showed type 2 muscle fibers atrophy with discrete inflammatory cells, predominantly lymphocytic in perivascular and interstitial locations. She was treated with prednisone and all the symptoms subsided after two months. PMID- 15122452 TI - Hypocalcemic myopathy without tetany due to idiopathic hypoparathyroidism: case report. AB - Myopathy due to idiopathic hypoparathyroidism is very unusual. We report on a 30 years-old man referred with complaints of sporadic muscle pain and mild global weakness for 10 years. His physical examination showed normal strength in distal muscle and slightly weakness in the pelvic and scapular girdles with no atrophy. Deep muscle reflexes were slightly hypoactive. Trousseau's and Chvostek's signs were absent. He had bilateral cataract and complex partial seizures. His laboratory tests showed decreased ionised and total calcium and parathyroid hormone and increased muscle enzymes. EMG and muscle biopsy was compatible with metabolic myopathy. After treatment with calcium and vitamin D supplementation he showed clinical, neurophysiological and laboratorial improvement. IN CONCLUSION: patients with muscle symptoms, even when non-specific and with normal neurological examination, should have serum calcium checked, as myopathy due to idiopathic hypoparathyroidism, even being rare, is treatable and easy to diagnose. PMID- 15122453 TI - Hypopituitarism and amenorrhea-galactorrhea syndrome caused by thrombosis of both internal carotid artery and giant intrasellar aneurysm: case report. AB - Giant intra and parasellar aneurysm with a spontaneous thrombosis of internal carotid artery is rare. We report the case of a 34 years old woman presenting a unique giant sellar and parasellar aneurysm associated with hypopituitarism and amenorrhea-galactorrhea syndrome. Computed tomographic scans and magnetic resonance images were suggestive of a sellar tumor with a cystic component. Digital cerebral angiography showed spontaneous thrombosis of a intrasellar and parasellar carotid artery aneurysm and left internal carotid artery in the neck. A transseptal endoscopic biopsy was done and confirmed a thrombosed aneurysm. No other surgical treatment was required in this patient but permanent endocrinological treatment was necessary. PMID- 15122454 TI - [Schwannoma of brachial plexus: report of two cases]. AB - Schwannomas, neurinomas or neurilemmomas are benign peripheral nerve tumors. The literature report some cases associated with neurofibromatosis 2. We report two cases of cervical schwannoma originating from the brachial plexus unassociated with neurofibromatosis. A 31-year-old woman presented with a mass in the right supraclavicular region, irradiating pain and distal tingling to percussion (Tinel's sign) for 6 months. And a 52-year-old woman presented with pain in the cervical region and right arm for one year. Both the patients underwent to a microsurgery with total resection of the lesion. Histology of the surgical specimen confirmed the diagnosis of schwannoma. Postoperatively, the patients had a good recovery. PMID- 15122455 TI - Multifocal osteoclastoma of the skull: case report. AB - We describe the case of a 35 years old man with a nonspecific complaint of a slow growing solid mass in the frontal region. Radiological exams evidenced two more lesions : in the superior and lateral walls of the orbit. Treated with total excision of the lesions and a cranioplastic procedure at the same act, with favorable outcome. Microscopic findings suggested giant cell tumor in the three lesions that was confirmed by immunohistochemical examination. PMID- 15122456 TI - Chronic subdural hematoma of the posterior fossa associated with cerebellar hemorrhage: report of rare disease with MRI findings. AB - Chronic subdural hematoma of the posterior fossa is an uncommon entity, and spontaneous lesions are very rarely described, occurring mostly during anticoagulation therapy. The association of the posterior fossa chronic subdural hematoma with spontaneous parenchymal hemorrhage without anticoagulation therapy was never related in the literature, to our knowledge. We describe a case of a 64 year-old woman who suffered a spontaneous cerebellar hemorrhage, treated conservatively, and presented 1 month later with a chronic subdural posterior fossa hematoma. PMID- 15122457 TI - [Primary low grade B-cell lymphoma MALT type of the duramater: case report]. AB - We describe a rare case of nodular and low grade non-Hodgkin malignant lymphoma, MALT type, with morphological and immunohistochemical features of lymphoplasmocytic (REAL classification), of immunophenotype of B lymphocytes and detected monoclonal of immunoglobulin kappa light chains, with low proliferation grade (<10%). The tumor was primitive of the left parietal duramater in a 36 years old woman, who presented neurological clinical symptoms four months before the surgery. After surgery, she was submitted to chemotherapy and radiotherapy with good results. PMID- 15122458 TI - [The Drew family of Walworth: one century from the first evaluation until the final diagnosis, Machado-Joseph disease]. AB - Autosomal dominant spinocerebellar ataxia (SCA) is an heterogeneous group of neurodegenerative diseases involving cerebellum and its connections. Several forms have already been described, and it seems the most common form of SCA observed among the many series of families described worldwide is SCA3 (Machado Joseph disease). SCA3 is characterized by a marked phenotypic expression with a wide spectrum of clinical findings including cerebellar ataxia, pyramidal and extrapyramidal (e.g. dystonia, parkinsonism), lower motor neuron syndrome and peripheral neuropathy. The Drew family of Walworth, England, has several affected members seen and described by famous neurologists including Gowers, Stewart, Collier, Kinnier-Wilson, Turner, Worster-Drought, Ferguson, Critchley, and Anita Harding from 1895 to our days. In fact, the final genetic diagnosis of this family, 100 years after its initial description, turned out to be SCA3. In this paper, we describe the full of twists and turns historical trajectory from the initial clinical description to the final genetic diagnosis. PMID- 15122459 TI - [Transcranial magnetic stimulation]. PMID- 15122460 TI - [Guidelines for myocardial revascularization surgery]. PMID- 15122461 TI - [Guidelines for surgery in heart valve diseases]. PMID- 15122462 TI - [Guidelines for surgery in aortic disease]. PMID- 15122463 TI - [International anatomical terminology: the new standard of nomenclature for the radiologist]. PMID- 15122464 TI - 3.0 T high-resolution MR imaging of carpal ligaments and TFCC. AB - PURPOSE: To determine the diagnostic value of 3.0 Tesla MRI for imaging carpal ligaments and triangular fibrocartilage complex (TFCC). Image quality of different optimized MRI sequences is evaluated for high resolution wrist anatomy. MATERIALS AND METHODS: Ten healthy volunteers were examined at 3.0 T and 1.5 T using following sequences: T1 SE, fat-saturated PD-/T2-TSE, TIRM, 3D T1/T2* DESS, 3D-CISS, 2D and 3D T2* MEDIC. Voxel size varied from 0.2 x 0.2 x 1.5 mm (2D sequences) to 0.33 mm (3) and 0.26 mm (3) (3D sequences). Image quality (signal to-noise-ratio, contrast-to-noise-ratio, artifacts) and carpal ligament/TFCC detection rate were judged by a score. The results obtained from the 3.0 T and 1.5 T devices were compared. RESULTS: With identical voxel size, image matrix and FOV, 3.0 T MRI provided significantly better image quality and ligament detection rates for all sequences in comparison with 1.5 T. The 2D and 3D MEDIC sequences yielded best image quality and detection rates. Excellent image quality and visualization of ligament structures by the fat-suppressed PD-TSE sequence were compromised by a relatively high susceptibility to pulsation and motion artifacts. T1 SE and 3D DESS sequences gave moderate image quality and allowed only partial differentiation between ligament structures. TIRM, T2-TSE and 3D CISS sequence proved to be unsuitable for examining ligaments at 3.0 T due to their poor image quality and detection rate. CONCLUSION: 3.0 T MRI of the wrist proved to be superior to 1.5 T MRI for high-resolution imaging of carpal ligaments and TFCC using 2D and 3D T2* MEDIC sequences. Clinical studies investigating ligament injuries or carpal instability are recommended for evaluating clinical relevance of high-resolution MRI of the wrist. PMID- 15122465 TI - [Imaging-based anatomy of the neck -- multi-slice CT and MRI]. AB - Purpose of this article is to provide accomplishments of the anatomy in the evaluation of pathologic findings is not a mandatory knowledge of every single anatomical structure, but rather a correct attribution of pathology to a single space of the neck. Only with an exact assignment to these spaces the differential diagnoses can be limited to a reasonable quantity to provide support for the referring physician in the ongoing treatment, especially for the planning of the various existing neck dissections. There are multifaceted descriptions of the anatomy of the head and neck region regarding an axial scanning with CT and MRI. Because of their intricacy it is difficult to use these classifications in a daily routine setting. Classifications with nearly as many spaces as anatomical structures provide no adequate support. Based on the various spaces of the head and neck region and additionally the imaging-based lymph node classification the aim of this work is to provide reasonable support in the daily clinical routine regarding the current literature and our own experience. PMID- 15122466 TI - [Imaging methods in the diagnosis and therapy of cystic echinococcosis]. AB - Imaging studies, ultrasonography, play a central role for the diagnosis and follow-up of cystic echinococcosis (hydatid disease) due to the non-specific clinical symptoms and still inadequate sensitivity and specificity of currently available serological tests. Due to the increasing number of people immigrating to central Europe from countries with a high incidence of cystic echinococcosis, cystic echinococcosis has become an important differential diagnosis of cystic lesions. The imaging modality to localize and stage the disease depends on the organs affected. Ultrasonography is the most important imaging technique to screen for abdominal lesions (more than 75 % of the cases). Therefore, an expert committee of the WHO Working Group on echinococcosis has recently suggested a standardized ultrasonographic classification of hepatic cystic echinococcosis. This classification proofs to be very useful for staging echinococcal cysts with respect to parasite activity. Ultrasonography is not only an excellent tool for the primary diagnosis and therapeutic decision but also for follow-up of patients treated for cystic echinococcosis. Indications for computed tomography or magnetic resonance tomography are restricted to extra abdominal disease, patients not suited for ultrasonography because of obesity or meteorism, complicated cysts and planning of surgery or interventional therapy. Apart from surgery three other treatment options are well established: (1) chemotherapy with albendazole or mebendazole, (2) percutaneous drainage and sterilization (PAIR) and (3) observation of inactive echinococcal stages ("watch and wait" approach). PMID- 15122467 TI - [Diagnostic value of preoperative contrast-enhanced MR imaging of the breast]. AB - PURPOSE: To evaluate preoperative contrast enhanced MR imaging in clinically, mammographically and/or ultrasonographically established breast cancer. MATERIALS AND METHOD: From September 1998 to August 1999, preoperative contrast-enhanced MR imaging of the breast was performed in 91 patients with lesions highly suggestive of malignancy (BIRADS IV and V) by clinical, mammographic, and/or ultrasonographic criteria. MR imaging findings were postsurgically correlated with other imaging, intraoperative and histopathologic results. RESULTS: Histopathologic analysis revealed 61 (66 %) malignant and 31 (34 %) benign lesions. In 63 (69 %) of the 91 investigated patients, MR mammographies were classified as tumor suspect and in the remaining 28 (31 %) cases as benign. The sensitivity, specificity and accuracy were 90 %, 67 % and 81 % for contrast enhanced MR imaging. Additional tumor manifestations (multifocal or multicentric disease, contralateral carcinoma) were found by MR imaging alone in 10 patients (11 %). CONCLUSION: Contrast-enhanced MR imaging may reveal unsuspected multifocal, multicentric or contralateral breast carcinoma that changes the surgical therapy if the intention is total tumor removal. The prognostic role of a potentially more radical surgical therapy on the basis of these findings is not clear. PMID- 15122468 TI - [Short-term CT findings after osteosynthesis of fractures of the vertebral spine]. AB - PURPOSE: Evaluation of a correlation between surgical stabilization of fractures of the vertebral spine and postoperative short-term CT findings with special attention to degenerative changes. MATERIALS AND METHODS: The pre- and post operative CT of 55 patients (43male, 12 female, age: 7 - 73 years, mean = 37.7 years) with traumatic fractures of the vertebral spine who underwent surgical stabilization using an internal fixator were analyzed retrospectively. The interval between surgical intervention and first postoperative CT control did not exceed one week. The interval to the second short-term follow-up CT ranged from 6 up to 24 months (mean: 10.3 +/- 2.7 months). The consensus reading of the short term CT by two experienced radiologists was compared to the pre- and immediately postoperative CT examinations and with the neurologic findings. Statistical analysis was done using the chi-square- or the Fisher's exact-test. RESULTS: No statistically significant correlation (p > 0.05) was found between the patient's age and the postoperative incidence of osteoporosis, spondylarthrosis, scoliosis, spondylolisthesis or disc protrusion after posttraumatic surgical stabilization using an internal fixator. Also, there was no significant correlation between the fracture's type or location, presence of polytraumatization, the technique of surgical intervention, and the incidence of postoperative degenerative changes in the neighboring segments. The thesis has not been verified (p > 0.05) that polysegment stabilization or prior surgical interventions at the vertebral spine with scar induction are responsible for a higher rate of degeneration. The only risk factor observed for postoperative scoliosis was the burst rotation fracture type C1.3 (AO-classification). CONCLUSIONS: No short-term acceleration of degenerative changes after surgical intervention in vertebral spine fractures was observed. However, due to the short-term interval between surgical intervention and follow-up CT (mean: 10.3 +/- 2.7 months), a final conclusion cannot yet be drawn. A follow-up CT seems to be indicated in patients with increasing complaints during this period. Thus, low grade complications can be diagnosed early. PMID- 15122469 TI - [Extended field-of-view sonography in Achilles tendon disease: a comparison with MR imaging]. AB - PURPOSE: To evaluate the role of real-time extended field-of-view sonography (EFOVS) in symptomatic Achilles tendon disease in comparison with MR imaging (MRI). MATERIALS AND METHODS: Twenty-three symptomatic tendons were examined by conventional grayscale sonography, EFOVS and MRI, which served as the gold standard. RESULTS: The median tendon thickness in MRI was 7.8 mm (IQR 3.1) and correlated significantly to the results of EFOVS (7.0 mm, IQR 2; r = 0.74, P < 0.01). In total, MRI detected 24 lesions in 18 tendons and EFOVS 21 hypoechoic lesions in 15 tendons, corresponding to a sensitivity of 87.5 % and specificity of 100 %. The additional usage of conventional grayscale sonography improved sensitivity to 95.8 %. The median distance of the largest lesion to the calcaneal tuberosity was 10.4 mm (IQR 3.4) in MRI and 8.5 mm (IQR 5.1) in EFOVS (r = 0.64; P < 0.05). The sensitivity and specificity of EFOVS for the detection of a peritendinitis were 63.6 % and 66.7 %, respectively. Corresponding values for the detection of a bursitis were 68.8 % and 28.6 %. The additional usage of conventional grayscale sonography improved the specificity to 85.7 %. CONCLUSION: The combination of EFOVS and grayscale sonography has the potential to challenge MRI as the preferred imaging method in diagnosing symptomatic Achilles tendon disease, especially with respect to saving time and cost and the absence of any contraindications. PMID- 15122470 TI - [Multislice-CT for structure analysis of trabecular bone - a comparison with micro-CT and biomechanical strength]. AB - OBJECTIVES: MS-CT (Multislice-Spiral-CT) has a higher spatial resolution compared to the SS-CT (Singleslice-CT). The purpose of this study was to investigate, if the higher spatial resolution of the MS-CT has advantages for structural analyses in the assessment of osteoporosis. MATERIAL AND METHODS: 20 cylindrical trabecular bone specimens (diameter 12 mm, length 15 - 20 mm) were harvested from formalin-fixed human thoracic spines. All specimens were examined by Micro-CT and quantitative, histomorphologic parameters were determined. Analogous structural parameters were calculated from the high-resolution images acquired by both MS- and SS-CT. Additionally, the BMD (bone mineral density) was measured by QCT (quantitative CT). The maximum compressive strength (MCS) was determined in a biomechanical test. The structural parameters were correlated with the histomorphologic parameters and with the MCS. RESULTS: The parameters bone fraction and trabecular separation correlated significantly in both MS- and SS-CT with the analogous parameters from Micro-CT (r (2) = 0.84, p < 0.01) and the MCS (r (2) = 0.81, p < 0.01). The highest correlation with the MCS was calculated using the trabecular number measured by MS-CT in the superior region near the endplate of the vertebra with the high-resolution kernel U90 u (r (2) = 0.85, p < 0.01). This correlation was significantly higher than the correlation between MCS and BMD (r (2) = 0.49, p < 0.01). CONCLUSION: Micro-CT- and MS-CT-determined structural parameters of the trabecular bone showed significant, high correlations. Thus, a characterisation of the trabecular structure seems to be possible. The biomechanical stability of the bone can also be predicted well. The structural parameters acquired by MS-CT show higher correlations with the MCS than the BMD or structural parameters determined by SS-CT do. In this study MS-CT was best suited to predict biomechanical strength of trabecular bone. PMID- 15122471 TI - [MR volumetry of the trigeminal nerve in patients with unilateral facial pain]. AB - PURPOSE: To assess whether MRI can detect atrophy of the trigeminal nerve in patients with trigeminal neuralgia. MATERIALS AND METHODS: A prospective MRI study was conducted in 39 patients (trigeminal neuralgia, trigeminal neuropathy, or atypical facial pain) and 25 volunteers. Using a coronal orientation (T1 Flash 3D; T2 CISS 3D), regions of interest were delineated in the cisternal part of the trigeminal nerve along the border of the nerve to calculate the volume of the nerve. The volume of the nerve was compared side-by-side in each patient (t-test, p < 0.05) and the volume difference compared between patients and volunteers. RESULTS: The volume of the compromised trigeminal nerve in patients with trigeminal neuralgia was lower than on the contralateral healthy side, with the difference between healthy and compromised side statistically significant (p < 0.05). In all other patients and in all volunteers, no significant difference was found between the volume of the healthy and compromised nerve. The volume difference between the healthy and compromised side in patients with trigeminal neuralgia was significantly higher (p < 0.05) than in all other patients and volunteers. CONCLUSION: Atrophy of the trigeminal nerve caused by a nerve-vessel conflict can be detected by MRI. Only patients with trigeminal neuralgia show this unilateral atrophy. Therefore, it is possible to demonstrate the result of the nerve-vessel conflict and to determine the consequences of such a conflict. PMID- 15122472 TI - [Determination of the renal blood flow in macro- and microcirculation by means of pulse inversion imaging]. AB - PURPOSE: To evaluate whether real-time and intermittent pulse inversion technology (PI) allows the analysis of blood flow in renal macro- and microcirculation. MATERIALS AND METHODS: The experiments were performed in a kidney perfusion phantom as an experimental model for the assessment of contrast replenishment in vascular regions of high flow velocity (medulla) and low flow velocity (cortex). During continuous infusion (0.03 ml/min) of Optison, contrast replenishment kinetics were assessed with intermittent PI at high emission power (MI: 1.3, with increasing trigger intervals) and with real-time PI at low emission power (MI: 0.09) at variable renal arterial blood flow (15 - 65 ml/min), using an HDI-5000 ultrasound unit (Philips Medical Systems). Regions of interest were placed in the major arteries of the medulla and the renal cortex to obtain replenishment curves of the macro- and microcirculation. Non-linear curve fitting was performed using the mathematical model y = A (1-e (-beta t)) with A as the parameter describing blood volume and beta as the parameter describing the speed of contrast replenishment. RESULTS: Replenishment curves could be obtained in all analyzed renal segments. For intermittent and real-time PI a strong linear correlation was found between renal arterial blood flow and A*beta (intermittent PI: cortex: R = 0.97; medulla: R = 0.98; real-time PI: cortex: R = 0.99; medulla: R = 0.96). The differences between the slopes of the regression lines (cortex: high power vs. low power, p = 0.844; medulla: high power vs. low power, p = 0.444) were not significant. CONCLUSION: Intermittent and real-time PI allows the assessment of renal blood flow in different vessel compartments. PMID- 15122473 TI - [High field MR imaging: magnetic field interactions of aneurysm clips, coronary artery stents and iliac artery stents with a 3.0 Tesla MR system]. AB - PURPOSE: To evaluate magnetic field interactions of commonly used biomedical implants at 3.0 Tesla. MATERIALS AND METHODS: Fourteen aneurysm clips designed for permanent placement in intracranial aneurysms, 19 coronary artery stents and 20 iliac artery stents were evaluated in an actively shielded compact 3.0 T MR system (Intera, Philips Medical Systems, Best, The Netherlands, length of magnet 1.57 m). The magnetic deflection forces (translational movement) were evaluated as follows: The implants were suspended by a fine string and placed in the magnet bore at the location of the maximum magnetic field gradient. The translational forces F (z) were calculated from the measured angle of deflection from the vertical axis. The magnetic field-induced torque (rotational forces) was evaluated as follows: Each implant was placed in the center of the magnetic bore parallel to the static magnetic field B0 (position 0 degrees ). Any possible displacement of the implant was noted on a millimeter scale and any torque qualitatively evaluated using a 5 point grading scale (0: no torque; + 4: very strong torque). The implant was turned in steps of 45 degrees, and the procedure was repeated to encompass a full 360 degrees rotation. RESULTS: In 52 of the 53 devices tested, the deflection force (deflection angle: range 0-21 degrees, translational force: range 0-3.8 mN) was less than the gravitational force (i.e., the implant's weight). These devices (n = 52/53) did not show any alignment to or rotation in the magnetic field at any of the various 45 degrees -increment positions corresponding to a qualitative torque evaluation of grade 0/4. One device (n = 1/53), an iliac artery stent made of stainless steel (Zenith, Cook, Monchengladbach, BRD), was found to have deflection forces (deflection angle 88 degrees translational force 299 mN) greatly exceeding the gravitational force as well as a pronounced torque (grade 4/4). CONCLUSION: Out of 53 biomedical implants evaluated for magnetic field interactions at 3.0 T, one iliac artery stent made of stainless steel was found to be potentially unsafe based on ASTM criteria. MR imaging at 3.0 Tesla may be performed safely in patients with any of the other 52 different implants evaluated in this study with respect to magnetic field translational attraction and torque. PMID- 15122474 TI - [Personal operator dose in invasive cardiology as a function of body height and tube angulation]. AB - PURPOSE: To map in an experimental setting of the local personal operator dose for 55 selected tube angulations as a function of body height above ground. MATERIALS AND METHODS: On an Alderson-Rando phantom representing the patient, we performed measurements of fluoroscopy scatter radiation ( micro Sv/h) at the operator's position, for the range of 20 - 200 cm body height, for all tube angulations in 30 degrees steps from right anterior oblique (RAO) 90 degrees to left anterior oblique (LAO) 90 degrees position, and for planes angulated cranially (+) and caudally (-) by 10 degrees, 20 degrees, 30 degrees, and 40 degrees, unless rendered unfeasible by geometric circumstances. RESULTS: Radiation exposure to the operator is lowest between postero-anterior (PA) 0 degrees and RAO 30 degrees angulation, and continuously increases by a factor of approx. 2 towards steep RAO, and to factors of 5 - 10 towards steep LAO views. Craniocaudal angulation at 30 degrees likewise generates personal dose levels 2 - 3 times as high. For all body heights and all LAO tube angulations, the corridor between 0 degrees - 10 degrees caudal angulation generates the least personal scatter dose, likewise irrespective of body height and craniocaudal tube angulations, the corridor between 0 degrees PA - 30 degrees RAO angulation. RAO angulations, however, being inverse to the respective 90 degrees LAO angulations, are generally 4 to 5 times less radiation extensive. Peak levels of the local personal dose vary from 160 cm body height for steep cranial LAO 90 degrees /30 degrees + views (3,500 microSv/h), to 50 cm for cranial PA 0 degrees /30 degrees + (400 micro Sv/h), and to > or = 170 cm (600 micro Sv/h) and < or = 40 cm (300 microSv/h) for steep cranial RAO 90 degrees /30 degrees + views. Caudal angulations generate slightly lower doses, with peak levels at 120 cm for LAO 90 degrees /30 degrees - views (3,000 microSv/h), at 50 cm for PA 0 degrees /30 degrees - views (300 micro Sv/h), and above 170 cm (900 micro Sv/h) and below 40 cm (500 microSv/h) for steep caudal RAO 90 degrees /30 degrees - views. CONCLUSION: The present experimental study on scatter radiation to the operator, as a function of body height and tube angulation, offers a representative data tool for all interventionists for use in invasive cardiology, to confirm the radiation safety of their favored coronary views, or to encourage less radiation intensive angulations. Moreover, it provides new knowledge about special risks for crucial body regions and enables effective radiation protection strategies. PMID- 15122475 TI - [Evaluation of experimental cartilage lesions with ultrahigh-resolution multi slice-CT in comparison to histology]. AB - PURPOSE: Histologic validation of ultrahigh-resolution multi-slice (MS)-CT for the evaluation of focal, experimental cartilage lesions with special regard to the subchondral bone. Testing of micro-CT ( micro CT) as alternative reference standard. METHODS: 32 experimental cartilage lesions in bovine patellae were imaged surrounded by air (MS-CT-air) and immersed in a contrast material solution (MS-CT-CM) with MS-CT (collimation 2 x 0,5 mm). After the micro CT (8 micro m voxelsite) examination in three specimen and histologic work-up of 29 specimen two radiologist graded the defects on MS-CT images in consensus (subchondral bone involvement yes or no) and results were compared to the results of histomorphometry and micro CT. RESULTS: The MS-CT-air and -CM had an accuracy of 94 % (30/32) and 88 % (28/32), respectively. MS-CT-air led to one false-positive (remaining cartilage: = 0,1 mm) and false-negative result, each. MS-CT-CM showed false-positive results if the remaining cartilage was < 0,3 mm thick (n = 4), i. e. showed subchondral bone involvement. Contrast-to-noise ratio was significantly higher in MS-CT-air compared to MS-CT-CM. micro CT yielded a clear depiction of cartilage defect depth in the three cases. CONCLUSION: MS-CT-air has a high accuracy in the depiction of focal cartilage defects. MS-CT-CM has a tendency to overestimate cartilage defect depth. micro CT could potentially serve as alternative reference standard to histology. PMID- 15122476 TI - [Flat-panel detector-based computed tomography: accuracy of experimental growth rate assessment in pulmonary nodules]. AB - PURPOSE: To determine the performance of an a-Si/CsJ flat-panel detector-based volumetric computed tomography (VCT) prototype in volumetry of synthetic nodules in a pulmonary phantom, and to assess VCT accuracy in the assessment of hypothetic tumor growth rates based on predefined tumor doubling times. METHODS: The true volumes of 50 synthetic nodules (diameter range = 1.36 - 5.34 mm) were determined and VCT volumetry was performed before and after isovolumetric deformation of the nodules. The percent measurement error (PMF) was calculated as the percent difference of the measured from the true volume. Based on the PMF, the minimum interval between two scans was determined that would be needed to depict tumor growth corresponding to the minimum number of required follow-up days (FUDs). Based on predefined tumor doubling times (VDT) FUDs were determined before and after nodule deformation. RESULTS: Measured volumes of undeformed and deformed nodules of 0.99 - 20.05 mm (3) differed significantly from corresponding true volumes (p = 0.002 - 0.004). The PMFs of these nodules significantly exceeded the values measured in larger nodules (p = 0.0001 - 0.0029). In addition, PMFs were significantly lower before than after deformation (1.33 - 7.14 % and 0.61 - 11.09 %, respectively; p = 0.002). For theoretical VDTs of 177 and 396 days, the calculated FUDs for detection of tumor growth were 19.1 and 42.7 days before deformation, and 30.2 and 67.6 days after deformation for nodules < 2 mm, respectively. CONCLUSION: VCT allows for accurate volumetry of smallest pulmonary nodules and may become a valuable clinical tool for depiction of tumor growth of even small lesions within very short scan intervals. PMID- 15122477 TI - [Fulminant invasive fungal sinusitis in immunosuppressed hosts -- pathognomic presentation in MRI]. PMID- 15122478 TI - [Pulmonary alveolar microlithiasis]. PMID- 15122479 TI - [Gallstone ileus]. PMID- 15122480 TI - [A newborn infant with large cervical cystic hygroma with hemangioma part]. PMID- 15122481 TI - [Intramural hematoma of the aorta with progression to a Stanford-A dissection]. PMID- 15122482 TI - ["Body stalk anomaly" (dysplasia umbilico-fetalis) -- an extremely rare malformation]. PMID- 15122484 TI - Parent-offspring conflict in the evolution of vertebrate reproductive mode. AB - We propose and evaluate the hypothesis that parent-offspring conflict over the degree of maternal investment has been one of the main selective factors in the evolution of vertebrate reproductive mode. This hypothesis is supported by data showing that the assumptions of parent-offspring conflict theory are met for relevant taxa; the high number of independent origins of viviparity, matrotrophy (direct maternal-fetal nutrient transfer), and hemochorial placentation (direct fetal access to the maternal bloodstream); the extreme diversity in physiological and morphological aspects of viviparity and placentation, which usually cannot be ascribed adaptive significance in terms of ecological factors; and divergent and convergent patterns in the diversification of placental structure, function, and developmental genetics. This hypothesis is also supported by data demonstrating that embryos and fetuses actively manipulate their interaction with the mother, thereby garnishing increased maternal resources. Our results indicate that selection may favor adaptations of the mother, the fetus, or both in traits related to reproductive mode and that integration of physiological and morphological data with evolutionary ecological data will be required to understand the adaptive significance of interspecific variation in viviparity, matrotrophy, and placentation. PMID- 15122485 TI - Adaptation, niche conservatism, and convergence: comparative studies of leaf evolution in the California chaparral. AB - Small leaves and low specific leaf area (SLA) have long been viewed as adaptations to Mediterranean-type climates in many species of evergreen woody plants. However, paleobotanical and floristic evidence suggests that in many cases these traits originated prior to the advent of the summer-drought climate regime. In this study, molecular phylogenies and ancestral state reconstructions were used to test the hypothesis of adaptive leaf evolution in 12 lineages of evergreen shrubs in the California chaparral. Across all lineages there was a small but significant shift toward lower SLA, but there were no trends in leaf size evolution. For individual lineages, adaptive changes were detected in only three cases for SLA and in one case for leaf size. Three of these cases of evolutionary change were observed in taxa derived from cool temperate ancestors (e.g., Heteromeles). In contrast, most lineages originating from subtropical ancestors exhibited relative stasis in leaf trait evolution (e.g., Ceanothus). The absence of change suggests that ancestors of chaparral taxa had already acquired appropriate traits that contributed to their success under Mediterranean type climates. These results illustrate how biogeographic history may influence patterns of trait evolution and adaptation and highlight the contribution of ecological sorting processes to the assembly and functional ecology of regional biotas. PMID- 15122486 TI - Similarity of mammalian body size across the taxonomic hierarchy and across space and time. AB - Although it is commonly assumed that closely related animals are similar in body size, the degree of similarity has not been examined across the taxonomic hierarchy. Moreover, little is known about the variation or consistency of body size patterns across geographic space or evolutionary time. Here, we draw from a data set of terrestrial, nonvolant mammals to quantify and compare patterns across the body size spectrum, the taxonomic hierarchy, continental space, and evolutionary time. We employ a variety of statistical techniques including "sib sib" regression, phylogenetic autocorrelation, and nested ANOVA. We find an extremely high resemblance (heritability) of size among congeneric species for mammals over approximately 18 g; the result is consistent across the size spectrum. However, there is no significant relationship among the body sizes of congeneric species for mammals under approximately 18 g. We suspect that life history and ecological parameters are so tightly constrained by allometry at diminutive size that animals can only adapt to novel ecological conditions by modifying body size. The overall distributions of size for each continental fauna and for the most diverse orders are quantitatively similar for North America, South America, and Africa, despite virtually no overlap in species composition. Differences in ordinal composition appear to account for quantitative differences between continents. For most mammalian orders, body size is highly conserved, although there is extensive overlap at all levels of the taxonomic hierarchy. The body size distribution for terrestrial mammals apparently was established early in the Tertiary, and it has remained remarkably constant over the past 50 Ma and across the major continents. Lineages have diversified in size to exploit environmental opportunities but only within limits set by allometric, ecological, and evolutionary constraints. PMID- 15122487 TI - Persistence and fluctuation of lateral dimorphism in fishes. AB - Two morphological types ("righty" and "lefty") have been discovered in several fish species and are referred to as a typical example of antisymmetry. It has been suggested, first, that this dimorphism (called laterality) is inheritable; second, that the frequencies of laterality in each species fluctuate around 0.5; and third, that predators mainly exploit prey of the opposite laterality; that is, lefty and righty predators prey on righties and lefties, respectively. The latter is defined as "cross predation"; the antonym "parallel predation" means predation within the same laterality. We hypothesized that cross predation drives alternation of the survival and reproductive advantages between two morphological types, leading to frequency-dependent selection that maintains the dimorphism. To investigate this, we constructed mathematical models of population dynamics of one prey/one predator systems and three-trophic-level systems with omnivory. Mathematical analysis and computer simulations explained the behavior of the laterality frequency in nature well, insofar as cross predation dominated over parallel predation. Furthermore, the simulations showed that when only one of the morphological types exists in a species, the other type can invade. This suggests that dimorphism is maintained in all interacting species. PMID- 15122488 TI - A putative mechanism for bog patterning. AB - The surface of bogs commonly shows various spatial vegetation patterning. Typical are "string patterns" consisting of regular densely vegetated bands oriented perpendicular to the slope. Here, we report on regular "maze patterns" on flat ground, consisting of bands densely vegetated by vascular plants in a more sparsely vegetated matrix of nonvascular plant communities. We present a model reproducing these maze and string patterns, describing how nutrient-limited vascular plants are controlled by, and in turn control, both hydrology and solute transport. We propose that the patterns are self-organized and originate from a nutrient accumulation mechanism. In the model, this is caused by the convective transport of nutrients in the groundwater toward areas with higher vascular plant biomass, driven by differences in transpiration rate. In a numerical bifurcation analysis we show how the maze patterns originate from the spatially homogeneous equilibrium and how this is affected by changes in rainfall, nutrient input, and plant properties. Our results confirm earlier model results, showing that redistribution of a limiting resource may lead to fine-scale facilitative and coarse-scale competitive plant interactions in different ecosystems. Self organization in ecosystems may be a more general phenomenon than previously thought, which can be mechanistically linked to scale-dependent facilitation and competition. PMID- 15122489 TI - Sympatric speciation by sexual selection: a critical reevaluation. AB - Several empirical studies put forward sexual selection as an important driving force of sympatric speciation. This idea agrees with recent models suggesting that speciation may proceed by means of divergent Fisherian runaway processes within a single population. Notwithstanding this, the models so far have not been able to demonstrate that sympatric speciation can unfold as a fully adaptive process driven by sexual selection alone. Implicitly or explicitly, most models rely on nonselective factors to initiate speciation. In fact, they do not provide a selective explanation for the considerable variation in female preferences required to trigger divergent runaway processes. We argue that such variation can arise by disruptive selection but only when selection on female preferences is frequency dependent. Adaptive speciation is therefore unattainable in traditional female choice models, which assume selection on female preferences to be frequency independent. However, when frequency-dependent sexual selection processes act alongside mate choice, truly adaptive sympatric speciation becomes feasible. Speciation is then initiated independently of nonadaptive processes and does not suffer from the theoretical weaknesses associated with the current Fisherian runaway model of speciation. However, adaptive speciation requires the simultaneous action of multiple mechanisms, and therefore it occurs under conditions far more restrictive than earlier models of sympatric speciation by sexual selection appear to suggest. PMID- 15122490 TI - Spatial scale and temporal component of selection in side-blotched lizards. AB - Spatial variation in selection has long been recognized as promoting population divergence and in maintaining genetic polymorphisms, but selection at a fine spatial scale is seldom measured directly. We analyzed spatial and temporal variation in selective regimes on egg size using long-term population data of the side-blotched lizard (Uta stansburiana). Juvenile survival rates varied between years at a small spatial scale that was reflected as a strong interaction between the local neighborhood level and year. Spatially and temporally variable selection acted jointly on egg mass, which presumably would facilitate the maintenance of high additive genetic variance for this trait. Local selection gradients calculated at the neighborhood level were significantly correlated with the annual global selection gradients calculated at the metapopulation level. However, there was substantial variance in these local selective regimes, which suggests that strong local selection could go undetected if the analysis was limited to the global level. We also investigated the degree of spatial synchronization among outcrop in local selection gradients. The degree of synchrony was higher among later-clutch hatchlings than among first-clutch hatchlings, and we suggest that more intense density- and frequency-dependent selection on egg size later in the season is responsible for this effect. PMID- 15122491 TI - Life-history consequences of investment in free-spawned eggs and their accessory coats. AB - The optimal trade-off between offspring size and number can depend on details of the mode of reproduction or development. In marine organisms, broadcast spawning is widespread, and external coats are a common feature of spawned eggs. Egg jelly coats are thought to influence several aspects of fertilization and early development, including the size of the target for sperm, fertilization efficiency, egg suspension time, polyspermy, embryo survival, and fecundity. These costs and benefits of investment in jelly result in trade-offs that can influence optimal reproductive allocation and the evolution of egg size. I develop an optimization model that sequentially incorporates assumptions about the function of egg coats in fertilization. The model predicts large variation in coat size and limited variation in ovum size under a broad range of conditions. Heterogeneity among spawning events further limits the range of ovum sizes predicted to evolve under sperm limitation. In contrast, variation in larval mortality predicts a broad range of optimal ovum sizes that more closely reflects natural variation among broadcast-spawning invertebrates. By decoupling physical and energetic size, egg coats can enhance fertilization, maintain high fecundity, and buffer the evolution of ovum size from variation in spawning conditions. PMID- 15122492 TI - Climate affects predator control of an herbivore outbreak. AB - Herbivore outbreaks and the accompanying devastation of plant biomass can have enormous ecological effects. Climate directly affects such outbreaks through plant stress or alterations in herbivore life-history traits. Large-scale variation in climate can indirectly affect outbreaks through trophic interactions, but the magnitude of such effects is unknown. On the California coast, rainfall in years during and immediately previous to mass lupine mortality was two-thirds that of years without such mortality. However, neither mature lupines nor their root-feeding herbivores are directly affected by annual variation in rainfall. By increasing soil moisture to levels characteristic of summers following El Nino/Southern Oscillation (ENSO) events, we increased persistence of a predator (the entomopathogenic nematode Heterorhabditis marelatus). This led to suppression of an outbreak of the herbivorous moth Hepialus californicus, indirectly protecting bush lupine (Lupinus arboreus). Our results are consistent with the marine-oriented Menge-Sutherland hypothesis (Menge and Sutherland 1987) that abiotic stress has greater effects on higher than on lower trophic levels. The mechanisms producing these results differ from those proposed by Menge-Sutherland, however, highlighting differences between trophic processes in underground and terrestrial/marine food webs. Our evidence suggests that herbivore outbreaks and mass lupine mortality are indirectly affected by ENSO's facilitation of top-down control in this food web. PMID- 15122493 TI - Timing and synchrony of ovulation in red deer constrained by short northern summers. AB - Iteroparous mothers often face a trade-off between further investments in current offspring at the expense of the start of the next reproductive cycle. In the strongly seasonal environments at northern latitudes, large herbivores are typically calving in early summer each year to get a long growth season and to hit peak protein levels of vegetation. Late-born offspring are more likely to die since they are smaller in autumn. Low female condition in autumn due to prolonged investment in current-year offspring may lower her ability to ovulate sufficiently early to get a good start for the calves the following spring. On the basis of autopsies of uteri from 10,073 red deer (Cervus elaphus), we show that ovulation was delayed as well as more synchronous with increasing population density. This suggests that ovulation beyond a certain date incurs some fitness costs. Ovulation occurs progressively earlier with increasing age up to around 13 yr of age, after which ovulation again occurs later. Low ovulation rates in young compared with prime-aged deer were correlated with late ovulation in the fall. Also, yearling groups with a low rate of ovulation (e.g., because of low weight) also ovulated later, and old senescent deer not calving the previous year ovulated less frequently and markedly later than those raising a calf. Our findings suggest, therefore, that mothers unable to ovulate before a certain date fail to do so altogether that year. PMID- 15122494 TI - Taxon richness and climate in angiosperms: is there a globally consistent relationship that precludes region effects? AB - We comment on a recent paper by Francis and Currie that reports a globally consistent relationship between plant family diversity in large geographic grid cells and factors in the physical environment. This relationship was interpreted as demonstrating local ecological determination of diversity, leaving little room for regional and historical effects. We suggest that using family richness to describe patterns of diversity reduces regional effects owing to the lower endemism of plants at the family compared with the species level. Regardless of the taxonomic level of analysis, we further point out that unless regional effects are tested explicitly, statistically significant differences in diversity between regions can be overlooked. Finally, even correlations between diversity and physical conditions of the environment can have primarily historical and evolutionary origins when lineages diversify within ecological zones of origin and spread more slowly to diverse environments. Thus, local ecological determinism should not have primacy over evolutionary-historical explanations for patterns of diversity between or within regions. Ecologists must evaluate patterns in local diversity within historical and geographic, as well as ecological, contexts. PMID- 15122496 TI - Butterfly selected lines explore the hormonal basis of interactions between life histories and morphology. AB - Hormonal mechanisms underlie many life-history traits and their interactions. We studied the role of ecdysteroids with regard to wing pattern and development time of the polyphenic butterfly Bicyclus anynana. Ecdysteroid titers and sensitivity to ecdysone injection were assayed for two-trait selected lines (ventral eyespot size and development time concurrently). These two traits are genetically and phenotypically coupled, having a common endocrinal basis. Two-trait selection had been applied both antagonistically (opposite the correlation) and synergistically (in the same direction as the correlation). Although selected lines had diverged most in eyespot size, the widest differences in timing of ecdysteroid titers were observed between the development time selection regimes; fast selected lines had an earlier hormonal increase after pupation than slow selected lines (even when corrected for differential pupal times). This endocrine peak was also earlier for females than for males. Furthermore, sensitivity to ecdysone injection as measured by a subsequent decrease in pupal time was significantly lower for slow selected lines than for fast or unselected lines. We conclude that the observed response in eyespot size to artificial selection must have been achieved via alteration of, or selection on, other developmental mechanisms, because the dynamics of the alternative, hormonal, pathway were dictated by development time selection. The developmental system is flexible enough to allow evolution in directions opposing the correlation between wing pattern and developmental time, and responses to selection are not constrained by a shared hormonal system. PMID- 15122497 TI - The growth/predation risk trade-off: so what is the mechanism? AB - Among damselflies in natural lakes, Ischnura species grow faster than coexisting Enallagma species, but Enallagma species have higher survival under predation than Ischnura species. This growth/predation risk trade-off apparently allows these taxa to coexist in ponds and lakes across the Holarctic. However, laboratory studies presented here show that the mechanism assumed by most theoretical and empirical studies to mediate this trade-off, namely activity simultaneously modulating foraging returns and predation risk, does not operate in this system. Ischnura verticalis larvae were more active than larvae of Enallagma species in a short-term behavioral experiment, which explains why Ischnura experiences greater mortality from predation. However, this greater activity did not translate into higher feeding rates. Ischnura verticalis and Enallagma species ate comparable amounts of food in both the short-term behavioral experiment and a longer feeding and digestion experiment. In spite of no difference in the amount of food ingested or assimilated, I. verticalis larvae grew faster than Enallagma larvae because they were better able to physiologically convert assimilated food into their own biomass in the presence of mortality threats. From these studies we understand the phenotypic mechanisms determining the antagonistic patterns of relative growth and survival between these two genera, but why these patterns exist remains unclear. PMID- 15122498 TI - A novel cost of R gene resistance in the presence of disease. AB - Resistance responses can impose fitness costs when pests are absent. Here, we test whether the induction of resistance can decrease fitness even in plants under attack; we call this potential outcome a net cost with attack. Using lines in which genetic background was controlled, we investigated whether susceptible Arabidopsis thaliana plants can outperform R gene resistant plants when infected with pathogens. For the R gene RPS2, there was a fitness benefit of resistance in the presence of intraspecific competition, but there was a net cost in the absence of competition: resistant plants produced less seed than susceptible plants even though infected with Pseudomonas syringae. This net cost was primarily due to overcompensation by susceptible plants, which occurred because of a developmental response to infection. For the R gene RPP5, there was no fitness effect of resistance without competition but a net cost when plants were infected with Peronospora parasitica in the presence of competition. This net cost was due to a reduction in the fitness of infected, resistant plants and complete compensation in susceptible plants. A spatially variable model suggests that a trade-off between net benefits and net costs with attack may help explain the persistence of individuals lacking R gene resistance to disease. PMID- 15122499 TI - Single-generation estimates of individual fitness as proxies for long-term genetic contribution. AB - Individual fitness is a central evolutionary concept, but the question of how it should be defined in empirical studies of natural selection remains contentious. Using founding cohorts from long-term population studies of two species of individually marked birds (collared flycatcher Ficedula albicollis and Ural owl Strix uralensis), we compared a rate-sensitive (lambdaind) and a rate-insensitive (lifetime reproductive success [LRS]) estimate of individual fitness with an estimate of long-term genetic fitness. The latter was calculated as the number of gene copies present in the population after more than two generations, as estimated by tracing genetic lineages and accounting for the fact that populations were not completely closed. When counting fledglings, rate insensitive estimates of individual fitness correlated better than rate-sensitive estimates with estimated long-term genetic contribution. When counting recruits, both classes of estimates performed equally well. The results support the contention that simple, rate-insensitive measures of fitness, such as LRS, provide a valid and good estimate of fitness in evolutionary studies of natural populations. PMID- 15122500 TI - Evolution restricts the coexistence of specialists and generalists: the role of trade-off structure. AB - Environmental variability and adaptive foraging behavior have been shown to favor coexistence of specialists and generalists on an ecological timescale. This leaves unaddressed the question of whether such coexistence can also be expected on an evolutionary timescale. In this article, we study the attainability, through gradual evolution, of specialist-generalist coexistence, as well as the evolutionary stability of such communities when allowing for immigration. Our analysis shows that the potential for specialist-generalist coexistence is much more restricted than originally thought and strongly depends on the trade-off structure assumed. We establish that ecological coexistence is less likely for species facing a trade-off between per capita reproduction in different habitats than when the trade-off acts on carrying capacities alone. We also demonstrate that coexistence is evolutionarily stable whenever it is ecologically stable but that in most cases, such coexistence cannot be reached through gradual evolution. We conclude that an evolutionarily stable community of specialists and generalists may be created only through immigration from elsewhere or through mutations of large effect. Our results highlight that trade-offs in fitness determining traits can have counterintuitive effects on the evolution of specialization. PMID- 15122501 TI - Predator mixes and the conspicuousness of aposematic signals. AB - Conspicuous warning signals of unprofitable prey are a defense against visually hunting predators. They work because predators learn to associate unprofitability with bright coloration and because strong signals are detectable and memorable. However, many species that can be considered defended are not very conspicuous; they have weak warning signals. This phenomenon has previously been ignored in models and experiments. In addition, there is significant within- and among species variation among predators in their search behavior, in their visual, cognitive, and learning abilities, and in their resistance to defenses. In this article we explore the effects of variable predators on models that combine positive frequency-dependent, frequency-independent, and negative frequency dependent predation and show that weak signaling of aposematic species can evolve if predators vary in their tendency to attack defended prey. PMID- 15122502 TI - Frequency and microenvironmental pattern of selection on plastic shade-avoidance traits in a natural population of Impatiens capensis. AB - The frequency and predictability of different selective environments are important parameters in models for the evolution of plasticity but have rarely been measured empirically in natural populations. We used an experimental phytometer approach to examine the frequency, predictability, and environmental determinants of heterogeneous selection on phytochrome-mediated shade-avoidance responses in a natural population of the annual plant Impatiens capensis. The strength and direction of selection on shade-avoidance traits varied substantially on a fine spatial scale. The shade-avoidance phenotype had high relative fecundity in some microsites but was disadvantageous in other microsites. Local seedling density proved to be a surprisingly poor predictor of microenvironmental variation in the strength and direction of selection on stem elongation in this study population. At least some of this unpredictability resulted from microenvironmental variation in water availability; the shade avoidance phenotype was more costly in dry microsites. Thus, environmental heterogeneity in resource availability can affect the relative costs and benefits of expressing shade-avoidance traits independent of local seedling density, the inductive environmental cue. Theory predicts that these conditions may promote local genetic differentiation in reaction norms in structured populations, as observed in I. capensis. PMID- 15122503 TI - Selection in modular organisms: is intraclonal variation in macroalgae evolutionarily important? AB - Theoretical and practical difficulties occur when defining the units of selection in modular organisms that grow by iteration of repeated parts (modules). Modules may become physically autonomous through fragmentation and may vary because of genetic variation arising in somatic cell lineages. Since cells destined for gamete production are not sequestered in early development, heritable variation and selection among asexual progeny are possible. We used the branching red macroalgae Delisea pulchra and Asparagopsis armata to test whether modules fulfill three fundamental criteria for units of selection: that they replicate, that they display heritable variation, and that selective agents distinguish among the variants. We detected significant phenotypic variation among modules for fitness-related traits (growth, secondary metabolite concentrations, and rates of tissue loss to herbivory) in each species and significant heritability estimates for secondary metabolite production and tissue loss to herbivory in D. pulchra. Variation in growth rate among A. armata modules was largely phenotypic with small but important estimates of genetic variation. Our results indicate that selection may indeed act on phenotypic variation among modules within individuals and that this process may effect evolutionary change within asexual lineages given sufficient genetic variation in the traits examined. PMID- 15122504 TI - Buffering of life histories against environmental stochasticity: accounting for a spurious correlation between the variabilities of vital rates and their contributions to fitness. AB - Life-history theory predicts vital rates that on average make large contributions to the annual multiplication rate of a lineage should be highly buffered against environmental variability. This prediction has been tested by looking for a negative correlation between the sensitivities (or elasticities) of the elements in a projection matrix and their variances (or coefficients of variation). Here, we show by constructing random matrices that a spurious negative correlation exists between the sensitivities and variances, and between the elasticities and coefficients of variation, of matrix elements. This spurious correlation arises in part because size transition probabilities, which are bounded by 0 and 1, have a limit to their variability that often does not apply to matrix elements representing reproduction. We advocate an alternative analysis based on the underlying vital rates (not the matrix elements) that accounts for the inherent limit to the variability of zero-to-one vital rates, corrects for sampling variation, and tests for a declining upper limit to variability as a vital rate's fitness contribution increases. Applying this analysis to demographic data from five populations of the alpine cushion plant Silene acaulis, we provide evidence of stronger buffering in the vital rates that most influence fitness. PMID- 15122505 TI - Spatial dynamics of specialist seed predators on synchronized and intermittent seed production of host plants. AB - Masting, the synchronized and intermittent seed production by plant populations, provides highly variable food resources for specialist seed predators. Such a reproductive mode helps minimize seed losses through predator satiation and extinction of seed predator populations. The seed predators can buffer the resource variation through dispersal or extended diapause. We developed a spatially explicit resource-consumer model to understand the effect of masting on specialist seed predators. The masting dynamics were assumed to follow a resource based model for plant reproduction, and the population dynamics of the predator were represented by a spatially extended Nicholson-Bailey model. The resultant model demonstrated that when host plants reproduce intermittently, seed predator populations go locally extinct, but global persistence of the predator is facilitated by dispersal or extended diapause. Global extinction of the predator resulted when the intermittent reproduction is highly synchronized among plants. An approximate invasion criterion for the predators showed that negative lag-1 autocorrelation in seeding reduces invasibility, and positive lag-1 cross correlation enhances invasibility. Spatial synchronization in seeding at local scale caused by pollen coupling (or climate forcing) further prevented invasion of the predators. If the predators employed extended diapause, extremely high temporal variability in reproduction was required for plants to evade the predators. PMID- 15122506 TI - Spatial flows and the regulation of ecosystems. AB - Spatial flows of materials and organisms across ecosystem boundaries are ubiquitous. Understanding the consequences of these flows should be a basic goal of ecosystem science, and yet it has received scant theoretical treatment to date. Here, using a simple, open, nutrient-limited ecosystem model with trophic interactions, we explore theoretically how spatial flows affect the functioning of local ecosystems, how physical mass-balance constraints interact with biological demographic constraints in the regulation of this functioning, and how failure to consider these constraints explicitly can lead to models that are ecologically inconsistent. In particular, we show that standard prey-dependent models for trophic interactions may lead to implausible outcomes when embedded in an ecosystem context with appropriate mass flows and mass-balance constraints. Our analysis emphasizes the need for integration of population, community, and ecosystem perspectives in ecology and the critical consequences of assuming closed versus open systems. PMID- 15122507 TI - Self-similarity and the species-area relationship. AB - Self-similar distributions of species across a landscape have been proposed as one potential cause of the well-known species-area relationship. The best known of these proposals is in the form of a probability rule for species occurrence. The application of this rule to the number of species occurring in primary well shaped rectangles within the landscape gives rise to a discrete power law for species-area relationships. However, this result requires a specific scheme for bisecting the landscape to generate the rectangles. Some additional, more general consequences of the probability rule are presented here. These include the result that the number of species in a well-shaped rectangle depends on its location, not just on its area. In addition, a self-similar landscape contains well-shaped rectangles that are, in fact, not self-similar. The probability rule in general produces testable predictions about how and where species are distributed that are independent of the power law. PMID- 15122508 TI - Self-similarity, the power law form of the species-area relationship, and a probability rule: a reply to Maddux. PMID- 15122509 TI - A general theory for the evolutionary dynamics of virulence. AB - Most theory on the evolution of virulence is based on a game-theoretic approach. One potential shortcoming of this approach is that it does not allow the prediction of the evolutionary dynamics of virulence. Such dynamics are of interest for several reasons: for experimental tests of theory, for the development of useful virulence management protocols, and for understanding virulence evolution in situations where the epidemiological dynamics never reach equilibrium and/or when evolutionary change occurs on a timescale comparable to that of the epidemiological dynamics. Here we present a general theory similar to that of quantitative genetics in evolutionary biology that allows for the easy construction of models that include both within-host mutation as well as superinfection and that is capable of predicting both the short- and long-term evolution of virulence. We illustrate the generality and intuitive appeal of the theory through a series of examples showing how it can lead to transparent interpretations of the selective forces governing virulence evolution. It also leads to novel predictions that are not possible using the game-theoretic approach. The general theory can be used to model the evolution of other pathogen traits as well. PMID- 15122510 TI - Meta-analysis of trade-offs among plant antiherbivore defenses: are plants jacks of-all-trades, masters of all? AB - On the basis of physiological and ecological costs of defense allocation, most plant defense theories predict the occurrence of trade-offs between resource investment in different types of antiherbivore defenses. To test this prediction, we conducted a meta-analysis of 31 studies published in 1976-2002 that provided data on covariation of different defensive traits in plant genotypes. We found no overall negative association between different defensive traits in plants; instead, the relationship between defensive traits varied from positive to negative depending on the types of co-occurring defenses. Evidence of trade-off was found only between constitutive and induced defenses. Therefore, to a large extent, plants appear to be jacks-of-all-trades, masters of all and may successfully produce several types of defense without paying considerable trade offs. Our survey thus provides little evidence that genetic trade-offs between defensive traits significantly constrain the evolution of multiple defenses in plants. PMID- 15122511 TI - CHEK2*1100delC and susceptibility to breast cancer: a collaborative analysis involving 10,860 breast cancer cases and 9,065 controls from 10 studies. AB - Previous studies of families with multiple cases of breast cancer have indicated that a frameshift alteration in the CHEK2 gene, 1100delC, is associated with an elevated frequency of breast cancer in such families, but the risk associated with the variant in other situations is uncertain. To evaluate the breast cancer risk associated with this variant, 10,860 breast cancer cases and 9,065 controls from 10 case-control studies in five countries were genotyped. CHEK2*1100delC was found in 201 cases (1.9%) and 64 controls (0.7%) (estimated odds ratio 2.34; 95% CI 1.72-3.20; P=.0000001). There was some evidence of a higher prevalence of CHEK2*1100delC among cases with a first-degree relative affected with breast cancer (odds ratio 1.44; 95% CI 0.93-2.23; P=.10) and of a trend for a higher breast cancer odds ratio at younger ages at diagnosis (P=.002). These results confirm that CHEK2*1100delC confers an increased risk of breast cancer and that this risk is apparent in women unselected for family history. The results are consistent with the hypothesis that CHEK2*1100delC multiplies the risks associated with susceptibility alleles in other genes to increase the risk of breast cancer. PMID- 15122512 TI - Mutated MESP2 causes spondylocostal dysostosis in humans. AB - Spondylocostal dysostosis (SCD) is a term given to a heterogeneous group of disorders characterized by abnormal vertebral segmentation (AVS). We have previously identified mutations in the Delta-like 3 (DLL3) gene as a major cause of autosomal recessive spondylocostal dysostosis. DLL3 encodes a ligand for the Notch receptor and, when mutated, defective somitogenesis occurs resulting in a consistent and distinctive pattern of AVS affecting the entire spine. From our study cohort of cases of AVS, we have identified individuals and families with abnormal segmentation of the entire spine but no mutations in DLL3, and, in some of these, linkage to the DLL3 locus at 19q13.1 has been excluded. Within this group, the radiological phenotype differs mildly from that of DLL3 mutation positive SCD and is variable, suggesting further heterogeneity. Using a genomewide scanning strategy in one consanguineous family with two affected children, we demonstrated linkage to 15q21.3-15q26.1 and furthermore identified a 4-bp duplication mutation in the human MESP2 gene that codes for a basic helix loop-helix transcription factor. No MESP2 mutations were found in a further 7 patients with related radiological phenotypes in whom abnormal segmentation affected all vertebrae, nor in a further 12 patients with diverse phenotypes. PMID- 15122513 TI - Fibroblast growth factor 20 polymorphisms and haplotypes strongly influence risk of Parkinson disease. AB - The pathogenic process responsible for the loss of dopaminergic neurons within the substantia nigra of patients with Parkinson disease (PD) is poorly understood. Current research supports the involvement of fibroblast growth factor (FGF20) in the survival of dopaminergic cells. FGF20 is a neurotrophic factor that is preferentially expressed within the substantia nigra of rat brain. The human homologue has been mapped to 8p21.3-8p22, which is within an area of PD linkage revealed through our published genomic screen. To test whether FGF20 influences risk of PD, we genotyped five single-nucleotide polymorphisms (SNPs) lying within the FGF20 gene, in a large family study. We analyzed our sample (644 families) through use of the pedigree disequilibrium test (PDT), the genotype PDT, the multilocus-genotype PDT, and the family-based association test to assess association between risk of PD and alleles, genotypes, multilocus genotypes, and haplotypes. We discovered a highly significant association of PD with one intronic SNP, rs1989754 (P=.0006), and two SNPs, rs1721100 (P=.02) and ss20399075 (P=.0008), located in the 3' regulatory region in our overall sample. Furthermore, we detected a haplotype (A-G-C-C-T) that is positively associated with risk of PD (P=.0003), whereas a second haplotype (A-G-G-G-C) was found to be negatively associated with risk of PD (P=.0009). Our results strongly support FGF20 as a risk factor for PD. PMID- 15122514 TI - Brief but efficient: acute HIV infection and the sexual transmission of HIV. AB - BACKGROUND: We examined whether viral dynamics in the genital tract during the natural history of acute human immunodeficiency virus type 1 (HIV-1) infection could explain efficient heterosexual transmission of HIV. METHODS: We measured HIV-1 concentration in blood and semen samples from patients with acute and long term HIV-1 infection. We explored the effect of changes in viral dynamics in semen on the probability of transmission per coital act, using a probabilistic model published elsewhere. RESULTS: Considered over time from infection, semen HIV-1 concentrations, in men with acute infection, increase and decrease in approximate parallel with changes occurring in blood. Modeling suggests that these acute dynamics alone are sufficient to increase probability of heterosexual transmission by 8-10-fold between peak (day 20 after infection, based on the model) and virologic set points (day 54 and later after infection). Depending on the frequency of coitus, men with average semen HIV-1 loads and without sexually transmitted diseases (STDs) would be expected to infect 7%-24% of susceptible female sex partners during the first 2 months of infection. The predicted infection rate would be much higher when either partner has an STD. CONCLUSIONS: Empirical biological data strongly support the hypothesis that sexual transmission by acutely infected individuals has a disproportionate effect on the spread of HIV-1 infection. Acute hyperinfectiousness may, in part, explain the current pandemic in heterosexual individuals. PMID- 15122515 TI - Rapid clearance of virus after acute HIV-1 infection: correlates of risk of AIDS. AB - OBJECTIVE: Our objective was to define early virologic and immunologic determinants of human immunodeficiency virus (HIV) type 1 disease progression among 22 case subjects with acute infection from the Trinidad Seroconvertor Cohort. METHODS: A linear segmented regression model was fitted to sequential quantitative virus load measurements. Parameters of virus kinetics during different phases of primary infection were correlated with clinical and immunologic end points, by use of Kaplan-Meier estimates and Cox regression. RESULTS: Ten individuals developed acquired immunodeficiency syndrome (AIDS) defining events. In univariate analysis, progression to AIDS was associated with rate of initial HIV clearance (P=.002), virus load during set point (P=.008), and CD4(+) cell count during steady state (P=.04). In the multivariate analysis, a rapid rate of initial clearance was the sole independent predictor of subsequent progression to AIDS and was associated with a 92% reduction in the risk of AIDS. The rate of initial clearance is inversely correlated with the number of early symptoms (r=-0.66; P=.0008). However, symptoms did not predict subsequent risk of AIDS. CONCLUSION: Among a subset of patients, rapid clearance of plasma HIV-1 after peak viremia is associated with lower viral set point, prolonged virus suppression before loss of virologic control, and decreased risk of AIDS. These findings are consistent with the hypothesis that effective immune responses during the earliest phase of infection are important determinants of the subsequent natural history. PMID- 15122516 TI - Identification of I50L as the signature atazanavir (ATV)-resistance mutation in treatment-naive HIV-1-infected patients receiving ATV-containing regimens. AB - Atazanavir (ATV) is a once-daily human immunodeficiency virus (HIV) protease inhibitor (PI) shown to be effective and well tolerated. ATV has a distinct resistance profile relative to other PIs, with susceptibility maintained against 86% of isolates resistant to 1-2 PIs. Clinical isolates obtained from PI-naive patients designated as experiencing virologic failure while receiving ATV containing regimens contained a unique isoleucine-to-leucine substitution at amino acid residue 50 (I50L) of the HIV-1 protease. The I50L substitution, observed in all isolates exhibiting phenotypic resistance to ATV, emerged in a variety of different backgrounds and was most frequently accompanied by A71V, K45R, and/or G73S. Viruses containing an I50L substitution were growth impaired, displayed ATV-specific resistance, and had increased susceptibilities (.05). No infection or failure of treatment was encountered. However, some patients in both groups had numbness or paresthesias of the cheek, upper lip, upper front gingiva, and teeth. These complaints were transient and lasted for several weeks. CONCLUSION: Based on our findings, it does not seem necessary to perform antrostomy at the inferior meatus, provided the patient has a patent osteomeatal complex and no anatomic abnormalities. PMID- 15122560 TI - A preliminary morphologic classification of the alveolar ridge after distraction osteogenesis. AB - PURPOSE: We sought to develop and evaluate a preliminary morphologic classification of the alveolar ridge after distraction osteogenesis. MATERIALS AND METHODS: Twelve patients (7 women and 5 men; mean age, 42.6 years; age range, 30 to 57 years) underwent a total of 17 alveolar ridge distractions before the placement of a total of 44 dental implants. Ridge bone morphology was evaluated and categorized at implant placement, with subsequent evaluation of the frequency and type of complications in each category. RESULTS: Four morphologic categories were identified, as follows. Category I consisted of wide alveolar rim and no bone defects; 7 (41.2%) of the 17 ridges were assigned to this category, and a total of 22 implants were placed with no complications. Category II consisted of wide alveolar rim, lateral bone surface concavity; 4 (24%) of the 17 ridges were assigned to this category, and a total of 8 implants were placed, with fenestration defects being the most frequent complication (2 of 8 implants [25%]). Category III consisted of narrow alveolar rim, lateral bone surface concavity; 5 (29.4%) of the 17 ridges were assigned to this category, and a total of 13 implants were placed, with dehiscence defects being the most frequent complication (4 of 13 implants [31%]). Category IV consisted of distraction transport segment forming a bridge, without bone formed beneath, necessitating guided bone regeneration; 1 (6%) of the 17 ridges was assigned to this category, and following bone regeneration 1 implant was placed, without complications. Subcategory D consisted of lingual deviation of the distraction axis, occurring in any of categories I to IV, and when severe requiring corrective osteotomy to free and reposition the transport segment and neoformed bone; 4 (24%) of the 17 ridges were assigned to this subcategory (ie, to subcategory ID, IID, IIID, or IVD); in 1 case, the deviation was severe, requiring corrective osteotomy; in this case 3 implants were placed, without complications. CONCLUSION: This preliminary morphologic classification of the postdistraction alveolar ridge effectively categorizes the variation observed in our patients and in our experience provides a useful basis for decision-making regarding implant placement. However, further studies are required to confirm the generality of this classification and incidences of complications in each category. PMID- 15122561 TI - Prophylaxis with cefazolin plus clindamycin in clean-contaminated maxillofacial surgery. AB - PURPOSE: Infections after maxillofacial surgery are usually due to aerobic and anaerobic gram-positive cocci and gram-negative bacilli. Various antimicrobials, including cephalosporins, beta-lactams/beta-lactamase inhibitors, aminoglycosides, lincosamides, and fluoroquinolones, have been tested for use for perioperative prophylaxis in maxillofacial surgery. However, the best regimen has not been determined. We tested the safety and the efficacy of clindamycin plus cefazolin as perioperative prophylaxis for patients undergoing major maxillofacial procedures. PATIENTS AND METHODS: Intravenous cefazolin and clindamycin in 3 doses were administered to 155 patients undergoing major maxillofacial procedures. After surgery, patients were monitored for the presence of infection and side effects. RESULTS: No patient experienced a fever or infection after surgery. No side effects related to these antibiotics were observed. CONCLUSIONS: The antibiotics used as prophylaxis in maxillofacial surgery should possess an adequate coverage against gram-positive aerobic and anaerobic cocci as well as gram-negative bacilli. Prophylaxis with cefazolin plus clindamycin in major maxillofacial seems safe and effective. PMID- 15122562 TI - Effect of restraint systems on maxillofacial injury in frontal motor vehicle collisions. AB - PURPOSE: Motor vehicle collisions (MVCs) are the leading cause of maxillofacial fractures. Additionally, maxillofacial injuries are the most common injury related to air bag deployment. We sought to characterize the occupant restraint system (seat belt and air bag) and collision characteristics associated with MVC related maxillofacial injuries. MATERIALS AND METHODS: The 1991-2000 National (United States) Automotive Sampling System Crashworthiness Data System (CDS) data files were used. The CDS is a national probability sample of passenger vehicles involved in police-reported tow-away MVCs. Analysis was limited to front seat occupants involved in frontal collisions of delta-V (estimated change in velocity) of greater than 15 km/hr. The risk of facial injury was calculated according to occupants' restraint use (unrestrained, seat belt only, air bag only, and seat belt and air bag combined) and compared using risk ratios (RRs) and associated 95% confidence intervals (CIs). RESULTS: Occupants restrained with a seat belt only (RR, 0.48; 95% CI, 0.40 to 0.57) or a seat belt and an air bag (RR, 0.83; 95% CI, 0.73 to 0.94) had a significantly reduced risk of any facial injury compared with completely unrestrained occupants. There was no association for those restrained with an air bag only (RR, 1.19; 95% CI, 0.82 to 1.73). A similar pattern of results was observed for moderate to severe facial injuries and for facial fractures. CONCLUSION: Seat belt use significantly reduces the risk of facial injury in frontal MVCs. Air bag use was not associated with the risk of facial injury. PMID- 15122563 TI - Perioperative neurosensory changes associated with treatment of mandibular fractures. AB - PURPOSE: The purpose of this study was to document perioperative neurosensory changes in the inferior alveolar nerve (IAN) after mandibular fracture treatment and to identify risk factors associated with these changes. MATERIALS AND METHODS: This was a prospective cohort study composed of patients treated for mandibular fractures. The primary study variable was the change between the postinjury/pretreatment IAN neurosensory examination score and the score after fracture reduction. Risk factors were categorized as demographic, anatomic, and treatment. Appropriate descriptive, bivariate, and multivariate statistics were computed. RESULTS: The cohort was composed of 61 patients with 97 fractured sides. Forty-seven fractures (49%) were located between the lingula and mental foramen, and 50 fractures (51%) were located distal to the mental foramen. Thirty nine fractures (41%) were displaced by 5 mm or more. Abnormal preoperative IAN neurosensory examinations were documented in 81% of the fractured sides. Fifty three fractures (54%) were treated by closed reduction and 44 (46%) by open reduction and internal fixation. In 82 fractures (85%), the IAN neurosensory score was unchanged or improved after treatment. In a multivariate model, preoperative neurosensory score, displacement, and treatment were associated with a statistically significant risk (P < or =.05) for postoperative deterioration of IAN sensation. CONCLUSION: Open reduction and internal fixation, fracture displacement of 5 mm or more, and a normal preoperative IAN neurosensory examination were associated with an increased risk for deterioration of the IAN neurosensory score after treatment of mandibular fractures. Additional studies are indicated to determine long-term IAN neurosensory function after mandibular fracture treatment and the factors that affect prognosis. PMID- 15122564 TI - Immediate reconstruction of frontal sinus fractures: review of 26 cases. AB - PURPOSE: The purpose of this study was to evaluate complications occurring after immediate reconstruction of severe frontal sinus fractures, including cases where the fracture was not limited to the anterior wall and also involved the posterior wall and/or sinus floor. PATIENTS AND METHODS: The records of twenty-six patients presently undergoing follow-up for frontal sinus fracture reconstruction were reviewed. Information regarding demographics, fracture characteristics and causes, associated facial fractures, use of grafts or implants, type of fixation used, nasofrontal duct management, use of antibiotics, and complications were noted. Patients were asked to return for clinical and radiographic follow-up to access late complications. RESULTS: The average age of patients with frontal fractures was 29.1 years and 92.3% were male. Mean follow-up was 3.6 years. The most common causes of fracture were motor vehicle accidents and physical aggression. All patients presented with comminuted and dislocated anterior wall fractures, 34.6% presented with posterior wall fractures, and 46% had sinus floor fractures. Complications occurred in 7 patients (26.92%) and included pneumoencephalus, frontal cutaneous fistula, frontal bone irregularity, and sinusitis. CONCLUSIONS: Frontal sinus reconstruction is a good procedure for immediate fracture treatment if there is not excessive comminution, dislocation, or instability of the posterior wall and if the frontonasal duct area is intact or can be repaired. Most complications result from incorrect indication for reconstruction. PMID- 15122565 TI - Tetracycline compound placement to prevent dry socket: a postoperative study of 200 impacted mandibular third molars. AB - PURPOSE: Our objective was to study whether the placement of intra-alveolar tetracycline prevents dry sockets or improves the postoperative period. PATIENTS AND METHODS: A comparative clinical study of the surgical removal of 200 impacted mandibular third molars is made, with particular reference to postextraction pain, inflammation, trismus, and the incidence of dry socket. In 50% of these cases, a pharmacologic preparation that includes tetracycline was placed in the socket after removal of the impacted molar. RESULTS: Dry socket was diagnosed in 4 cases (2%), with no relation to intra-alveolar tetracycline placement being observed. The patients who were administered intra-alveolar tetracycline had less pain and trismus and consumed fewer analgesics than the patients who received no such treatment, although statistical significance was not reached. CONCLUSIONS: The intra-alveolar placement of tetracycline compound after the surgical removal of impacted mandibular third molars did not affect the incidence of dry socket. PMID- 15122566 TI - Effect of exposed inferior alveolar neurovascular bundle during surgical removal of impacted lower third molars. AB - PURPOSE: In this prospective study, we sought to determine the incidence of inferior alveolar nerve (IAN) paresthesia in patients with an exposed IAN bundle seen intraoperatively. PATIENTS AND METHODS: We included consecutive patients undergoing third molar surgery in whom an exposed IAN bundle was seen in the third molar socket intraoperatively. Data recorded from patients included radiographic findings, variations in surgical method, intraoperative findings, complications, and postoperative sequelae. Patients were reviewed 1 week after surgery and evaluated subjectively and objectively to determine the incidence of paresthesia when the IAN bundle was exposed. Patients with paresthesia were followed for 2 years or until it resolved. RESULTS: An exposed IAN bundle was seen in 192 operation sites in 170 patients over a 5-year period, of which 166 patients with 187 operation sites were included in this study. Thirty-eight operation sites (20.3%) showed paresthesia at 1 week after surgery (95% confidence interval, 14.5% to 26.1%); 20 sites (15.0%) showed abnormal objective assessment results. By 3 months from surgery, 57.9% of nerves had recovered to normal sensation, 65.8% of nerves recovered by 6 months, and 71.1% of nerves recovered by 1 year. Eight patients were lost to follow-up. Logistic regression showed that the operator, male gender, older age, and root curvature were significant risk factors for paresthesia. CONCLUSIONS: Sighting an exposed intact IAN bundle during third molar surgery indicates its intimate relationship with the third molar and carries a 20% risk of paresthesia, with a 70% chance of recovery by 1 year from surgery. PMID- 15122568 TI - Mass in the masseter muscle. PMID- 15122567 TI - Reconstruction of mandibular defects with autologous tissue-engineered bone. AB - PURPOSE: Maxillofacial reconstructive procedures often require bone graft harvesting, which results in donor site morbidity; the use of tissue-engineered bone would eliminate this problem. In this study, a novel scaffold design and new fabrication protocol were used to produce autologous tissue-engineered constructs (scaffolds seeded with cells) to reconstruct segmental mandibular defects in a minipig model. MATERIALS AND METHODS: Porcine mesenchymal stem cells were isolated from the ilium. They were expanded in culture and seeded onto poly-dl lactic-coglycolic acid scaffolds. The constructs were placed in a bioreactor and incubated for 10 days in medium and osteogenic supplements. Four full-thickness bony defects (2 x 2 cm) were created in the same pig's mandible. The constructs (n = 2) were placed into 2 of the defects as autologous grafts. One unseeded scaffold and 1 empty defect served as controls. At 6 weeks postimplantation, the pig was sacrificed, the mandible was harvested, and the grafted sites were evaluated by clinical, radiographic, and histologic methods. RESULTS: The construct-implanted defects appeared to be filled with hard tissue resembling bone, whereas controls were filled with fibrous tissue. Radiographically, the tissue-engineered constructs were uniformly radiodense with bone distributed throughout. The interface between native bone and constructs was indistinct. Complete bone ingrowth was not observed in control defects. Osteoblasts, osteocytes, bone trabeculae, and blood vessels were identified throughout the defects implanted with constructs. CONCLUSION: This "proof-of-principle" study indicates that porcine mandibular defects can be successfully reconstructed by in vitro cultured autologous porcine mesenchymal stem cells on a biodegradable polymer scaffold with penetration of bone and blood vessels. PMID- 15122569 TI - Management of mentalis muscle dysfunction after advancement genioplasty: a case report. PMID- 15122570 TI - Anterior bilateral presentation of Stafne defect: an unusual case report. PMID- 15122571 TI - The use of argon beam coagulation to control hemorrhage: a case report and review of the technology. PMID- 15122572 TI - Mitek anchors in facial injury: an approach for soft tissue flap fixation. PMID- 15122573 TI - A novel gene mutation in an infant with cranial dysmorphology and orbital maxillary hypoplasia. PMID- 15122574 TI - Obstructive parotitis after rhytidectomy: case report. PMID- 15122575 TI - The adjustable vector deep plane midface lift. PMID- 15122577 TI - Science, the UICC and global cancer control. PMID- 15122576 TI - Reconstruction of a partial maxilla with a combination of autologous bone particles and a microtitanium mesh tray covered by a forearm flap. PMID- 15122578 TI - Modified adenoviruses for cancer gene therapy. AB - Adenoviral gene therapy is an exciting novel approach for treating cancers resistant to currently available therapies. However, currently there is little evidence supporting significant clinical benefits with replication-incompetent adenoviruses. Recent data suggest that expression of the primary receptor, the coxsackie-adenovirus receptor (CAR), may be highly variable on tumor cells, resulting in resistance to infection. Consequently, various strategies have been evaluated to modify adenovirus tropism in order to circumvent CAR deficiency, including retargeting complexes or genetic capsid modifications. To improve tumor penetration and local amplification on the antitumor effect, selectively oncolytic agents, i.e., conditionally replicating adenoviruses, have been constructed. Infection of tumor cells results in replication, oncolysis and subsequent release of the virus progeny. Normal tissue is spared due to lack of replication. This review focuses on the various modifications that have been investigated for improving the antitumor effect of adenoviral gene therapy. PMID- 15122579 TI - Zerumbone, a sesquiterpene in subtropical ginger, suppresses skin tumor initiation and promotion stages in ICR mice. AB - We recently showed that zerumbone, a sesquiterpene found in subtropical ginger, suppresses colonic tumor marker formation in rats and induces apoptosis in colon cancer cell lines. In our present study, the anti-tumor initiating and promoting activities of zerumbone in mouse skin were evaluated using a conventional 2-stage carcinogenesis model. A single topical pretreatment to mouse skin (2 micromol) 24 hr before application of dimethylbenz[a]anthracene (0.2 micromol) markedly suppressed tumor incidence by 60% and the number of tumors by 80% per mouse. Repeated pretreatment (16 nmol) twice weekly during the post-initiation phase reduced the number of 12-O-tetradecanoylphorbol-13-acetate (TPA, 1.6 nmol) induced tumors by 83% as well as their diameter by 57%. Multiple reverse transcriptase (RT) PCR experiments revealed that zerumbone (2 micromol) enhanced the mRNA expression level of manganese superoxide dismutase, glutathione peroxidase-1, glutathione S-transferase-P1 and NAD(P)H quinone oxidoreductase in the epidermis, but not that of cytochrome p450 1A1 or 1B1. Further, it diminished TPA-induced cyclooxygenase-2 protein expression and phosphorylation of extracellular signal-regulated kinase 1/2, while pretreatment(s), in either the priming or activation stage or both, reduced double TPA application-induced hydrogen peroxide formation and edema induction by 29% to 86%, respectively. Histologic examination revealed that pretreatment(s) with zerumbone suppressed leukocyte infiltration and reduced proliferating cell nuclear antigen-labeling indices. Together, our results indicate that zerumbone is a promising agent for the prevention of both tumor initiating and promoting processes, through induction of anti-oxidative and phase II drug metabolizing enzymes as well as attenuation of proinflammatory signaling pathways. PMID- 15122580 TI - Low-fat milk promotes the development of 7,12-dimethylbenz(A)anthracene (DMBA) induced mammary tumors in rats. AB - Commercial cow milk contains considerable amounts of estrogens. Our study assessed the effect of commercial low-fat milk on the development of 7,12 dimethylbenz(a)anthracene (DMBA)-induced mammary tumors in rats. Eighty 6-week old female Sprague-Dawley rats received a single oral dose of 5 mg DMBA. Twenty four hours later, the animals were divided into 4 groups of 20 animals each and given 1 of 4 test solutions for 20 weeks as their drinking liquid: low-fat (1%) milk (M), artificial milk (A), estrone sulfate solution (0.1 microg/ml, E), or tap water (W). The artificial milk was formulated to supply essentially the same calories as the milk. The low-fat milk contained 378 pg/ml estrone sulfate. Tumor incidence, the cumulative number of tumors and the sum of tumor diameters were higher in the M and E groups than in the A or W groups. Overall, the development of mammary tumors was in the order: M = E > A = W. Whereas the plasma 17beta estradiol concentration in the M group was the 2nd highest after the E group, the plasma level of insulin-like growth factor (IGF-I) was significantly higher in the M group than in the other 3 groups. In conclusion, commercially available low fat milk promotes the development of DMBA-induced mammary tumors in rats. The degree of the promotion is almost comparable to that of 0.1 microg/ml estrone sulfate. The high estrogen content in the milk may be responsible for the promotional effects, acting in concert with other hormones such as IGF-I. PMID- 15122581 TI - Cellular interactions in the tropism of prostate cancer to bone. AB - At autopsy >or=80% of prostate cancers have established macrometastases in marrow containing bone. The mechanism(s) to explain this remarkable level of bone involvement remain to be elucidated. We examined the adhesive and invasive behavior of prostate cancer cells to osteoblastic and human bone marrow endothelial cells (HBME-1) in an attempt to explain the tropism of prostate cells for bone. We found an inverse relationship between adhesion and prostate cell tumorigenicity and metastatic potential. Relative cell adhesion of P69 between cell lines was 1.74-fold (95% confidence interval [CI] = 1.15-2.64) and 1.58-fold (95% CI = 0.94-2.68) greater at 1 hr and 2 hr, respectively, than LNCaP that was essentially equivalent to C4-2 cells when using an osteoblastic cell line, D1 as the substrate. Similar results were acquired when HBME-1 were used as substratum. There was a marked increase in adhesion of the poorly tumorigenic cell line P69 as compared to the cancer cells to HBME-1. P69 adhesion was 2.78-fold (95% CI = 1.87-4.84) and 2.0-fold (95% CI = 1.43-2.80) greater at 1 hr and 2 hr, respectively when compared to LNCaP or C4-2 cells. D1 cells, a bone homing osteoblastic precursor, behaved contrary to the metastatic, bone-colonizing C4-2 cell line and bound best to other bone cells but not as well as a non-homing fetal bone marrow-derived cell line, D2. Invasion of prostate cancer cells through HBME-1 lawns was examined at 8 hr and 16 hr. In contrast to the adhesion studies, the invasion of the more aggressive C4-2 cells was 3.46-fold (95% CI = 1.18-10.17) and 2.65-fold (95% CI = 1.26-5.56) greater at 8 hr and 16 hr, respectively than LNCaP cells. Similarly, LNCaP cell invasion was 1.73-fold (95% CI = 0.69-4.37) and 2.35-fold (95% CI = 1.41-3.93) greater at 8 hr and 16 hr, respectively than P69 cells at the invasion of HBME-1 monolayers. At 8 hr, C4-2 cells had 6.0-fold (95% CI = 2.63,13.65) higher invasive potential than P69 cells. Phage display biopanning of LNCaP cells versus C4-2 cells in vitro using 4 separate techniques repeatedly identified the same peptide in support of minimal cell surface changes associated with the ability of C4-2 cells to metastasize to bone. As integrins are vital to cell adhesion and migration, we examined the integrin subunit expression in the prostate cell lines. The expression of integrin subunits is much higher in the nontumorigenic cell line, P69, whereas the differences in integrin expression between LNCaP and C4-2 are negligible. Only alpha(2) and beta(5) integrin subunits increase from LNCaP to C4-2. Given that C4-2 cells spontaneously metastasize to bone in vivo and LNCaP cells do not, these studies imply that the ability of a metastatic prostate cancer cell to colonize the bone is not completely dependent upon the ability of the cancer cell to adhere to either osteoblastic cells or to the bone marrow endothelial cell lining. Therefore, the initial interaction between the bone endothelium or stroma and prostate cells is not accurately referred to as a tropic or homing response. The invasion assay results indicate that the invasive potential of the cell more accurately reflects the bone colonizing potential of a prostate cancer cell. It is likely that bidirectional paracrine interactions, subsequent to marrow adhesion, between prostate cancer cells and the bone microenvironment are what determine the successful colonization of the bone by prostate cancer cells. Further, functional changes in surface proteins that are involved in invasion are likely to occur without major changes in levels of cell surface protein expression. Functional integrin association, substratum usage and outside in signaling are more likely to predict metastatic behavior. PMID- 15122582 TI - Lactosylsulfatide expression in hepatocellular carcinoma cells enhances cell adhesion to vitronectin and intrahepatic metastasis in nude mice. AB - Lactosylsulfatide (SM3), one of the major sulfated glycolipids, has been reported to be involved in cellular adhesion. Yet, its specific function has not been well understood in tumor biology, especially in the process of metastasis. We analyzed expression levels of sulfatide on HCC cells with different metastatic potentials and found that levels were correlated with metastatic potential. Next, the cerebroside sulfotransferase (CST) (EC2.8.2.11) gene, which synthesizes SM3 as well as galactosylsulfatide (SM4), was transfected into the HCC line Hep3B. Cell surface expression of SM3 was confirmed by thin-layer chromatogram immunostaining and flow-cytometric analyses. SM3-expressing Hep3B cells showed elevated expression of integrin alphaVbeta3 and higher adhesive ability to vitronectin compared to mock cells. Furthermore, SM3 expression promoted intrahepatic metastasis in nude mice. Thus, SM3 may play an important role in the metastasis of HCC cells by causing the interaction of integrin alphaVbeta3 with vitronectin. PMID- 15122583 TI - MRP1 and glucosylceramide are coordinately over expressed and enriched in rafts during multidrug resistance acquisition in colon cancer cells. AB - Previously we have described a novel multidrug-resistant cell line, HT29(col), which displayed over expression of the multidrug-resistance protein 1 (MRP1) and an altered sphingolipid composition, including enhanced levels of glucosylceramide (GlcCer; Kok JW, Veldman RJ, Klappe K, Koning H, Filipeanu C, Muller M. Int J Cancer 2000;87:172-8). In our study, long-term screening revealed that, during colchicine-induced acquisition of multidrug resistance in a new HT29(col) cell line, increases in GlcCer occurred concomitantly with upregulation of MRP1 expression. Both MRP1 and GlcCer were found enriched in Lubrol-insoluble membrane domains. The expression of MRP1 and GlcCer were tightly correlated, as indicated also by a reversal of both at the later stage of colchicine consolidation. Resistance to colchicine was determined by MRP1, while glucosylceramide synthase (GCS) did not contribute: 1). Resistance was fully inhibited by MK571. 2). GCS expression and activity were not upregulated in HT29(col) cells. 3). Inhibition of GCS did not affect MRP1-mediated efflux function or sensitivity to colchicine. Instead, overall sphingolipid metabolism was upregulated through an increased rate of ceramide biosynthesis. In conclusion, upregulation of MRP1 occurs in concert with upregulation of GlcCer during multidrug-resistance acquisition, and both are enriched in rafts. The increased GlcCer pool does not directly modulate MRP1 function and cell survival. PMID- 15122584 TI - Linear relationship between Wnt activity levels and apoptosis in colorectal carcinoma cells exposed to butyrate. AB - We have reported that butyrate, a fatty acid produced by dietary fiber that induces cell cycle arrest, differentiation and/or apoptosis in colorectal carcinoma (CRC) cells in vitro, modulates Wnt activity in 2 CRC cell lines (Bordonaro et al., Int. J. Cancer, 2002; 97:42-51). Our study determines how changes in the levels of Wnt activity induced by butyrate relate to the effects of butyrate on apoptosis, cell cycle arrest and differentiation of CRC cells. In 10 human CRC cell lines a direct relationship was shown between apoptosis and butyrate-induced increase in Wnt activity, as well as between suppressed clonal growth and increased Wnt activity. No correlation existed between butyrate induced increase in Wnt activity and differentiation. The direct relationship between apoptosis and Wnt activity was supported by analyses of DLD-1 and HCT-116 cells expressing a dominant negative form of Tcf4, and therefore, with repressed Wnt activity, as well as by measuring the ratio of apoptotic to live cells in flow cytometry-sorted cell fractions with high and low Wnt activity. Novel flow cytometric methodology was utilized to show that butyrate differentially increases the number of cells with Wnt activity in different CRC cell lines. Thus, CRC cell lines in which butyrate upregulated Wnt activity to relatively high levels were most susceptible to the apoptotic effects of butyrate, whereas cell lines in which butyrate modestly modulated Wnt activity were less affected. PMID- 15122585 TI - Overexpression of cdk4/cyclin D1, a possible mediator of apoptosis and an indicator of prognosis in human primary lung carcinoma. AB - The relation between expression of cell cycle-regulator molecules and apoptosis was examined in surgical specimens and cultured human lung carcinoma cell lines. Immunohistochemical analysis for 133 cases revealed 2 types of staining pattern. The first group consisted of 95 cases (71.4%) characterized by apoptotic cells showing intensely positive staining for cdk4 and cyclin D1 but negative for other proteins (type A). In the second group (type B), comprising 38 cases (28.6%), apoptotic cells exhibited intense positive staining for any cyclins and cdks. Most of the latter cases had lost expression of Rb protein. When tumor cells retrieved from paraffin-embedded tissue were examined by flow cytometry, higher proportions of cells expressing only cdk4 or cyclin D1 in type A cases and of cells expressing any cyclin or cdk in type B cases showed a subdiploid DNA content. In survival analysis using the LI of apoptotic cells and cyclin/cdk positive cells, the high-apoptosis/high-cyclin D1 group showed the poorest prognosis. Furthermore, forced overexpression of only cdk4 or cyclin D1 induced apoptosis in cultured cells with normal Rb protein, whereas overexpression of any cyclin or cdk induced apoptosis in cells defective for Rb protein. In conclusion, upregulation of cdk4/cyclin D1 may be a primary and critical factor in induction of apoptosis in human lung carcinomas in vivo. Moreover, inactivation of Rb protein renders cells more prone to apoptosis by abnormal expression of any cell cycle protein. PMID- 15122586 TI - Analysis of HIC-1 methylation and transcription in human ependymomas. AB - Ependymomas are among the most common brain tumors in children. They develop from ependymal cells lining the ventricular system of the CNS. Previous studies have demonstrated a significant rate of allelic loss at chromosome 17p13.3. The HIC-1 putative tumor-suppressor gene, which exhibits hypermethylation and loss of expression in various tumor entities including medulloblastomas and gliomas, maps to the affected region. In the present study, we analyzed HIC-1 in ependymomas. Therefore, we applied methylation-specific PCR of the 5'-untranslated region as well as of a central region of HIC-1 and bisulfite sequencing to determine the methylation status in 52 ependymomas of different histologic subtypes, grades and locations. In addition, we used a competitive RT-PCR approach for sensitive assessment of HIC-1 transcripts. Hypermethylation of at least one of the 2 analyzed regions was found in 43/52 (83%) cases. There was a significant correlation between hypermethylation of HIC-1 and nonspinal localization (p = 0.019) as well as age. Of 27 ependymomas, 22 (81%) showed absent or low expression of HIC-1. The elevated methylation of HIC-1 in nonspinal ependymomas supports the hypothesis that spinal and nonspinal ependymomas represent genetically distinct entities. PMID- 15122587 TI - Definition of candidate low risk APC alleles in a Swedish population. AB - Many families experience an apparently inherited increased risk of colorectal cancer (CRC) similar to the known syndromes familial adenomatous polyposis (FAP) and hereditary nonpolyposis colorectal cancer (HNPCC). Besides these high-risk syndromes, approximately 10% of all CRC cases come from families with 2 affected 1st-degree relatives, and even 1st-degree relatives to a single case of CRC are at increased risk. Risk subjects from these families frequently show polyps at colonoscopy, which suggests the APC gene as a good candidate susceptibility gene for these attenuated polypotic syndromes. We used the sensitive DHPLC technique to search for possible predisposing germline mutations in the entire APC gene in 91 risk subjects from these high- and low-risk syndromes with unknown predisposing genes. Most exons were also screened for mutations in 96 normal controls and 96 colorectal cancer cases. In our study we probably have identified the most common APC variants in a Swedish population. Among 30 germline variants identified, 1 clearly pathogenic nonsense mutation and 11 putative pathogenic variants (10 missense and one 3' UTR) were found in 20 index patients (22%). Twelve silent as well as 5 intronic variants were considered nonpathogenic. Two of the missense variants found here, E1317Q and D1822V, have previously been related to a difference in risk of colorectal cancer. One variant, 8636C>A, located within the 3' UTR region of the APC gene, was suggested to constitute an additional low risk allele with a similar relative risk as the Jewish I1307K mutation (OR = 1.8; 95% CI, 0.96-3.40). The question of whether all the other variants confer an increased colorectal cancer risk warrants future large association studies. PMID- 15122588 TI - Germline mutation and large deletion analysis of the CDKN2A and ARF genes in families with multiple melanoma or an aggregation of malignant melanoma and breast cancer. AB - The CDKN2A/ARF genes have been associated with increased risk of malignant melanoma (MM) in families with multiple members affected by disease and in families characterized by the constellation of breast cancer and MM. The exact contribution of CDKN2A/ARF to disease risk remains poorly characterized, especially in diverse populations. In this report, the contribution of CDKN2A/ARF germline mutations and large rearrangements to disease in Polish familial MM (FMM) and aggregations of breast cancer and MM were assessed using a strategy that included genomic sequencing, restriction fragment length polymorphism and multiplex ligation-dependent probe amplification. We examined 16 FMM cases (group 1), 44 MM probands with a cancer family aggregation (CFA) that included at least one breast cancer (group 2) and 22 breast cancer probands with CFA and MM (group 3). The results revealed a paucity of mutations in CDKN2A/ARF, suggesting that in the Polish population this gene does not contribute significantly to either FMM or MM within the context of CFA. PMID- 15122589 TI - Serologic detection of diffuse large B-cell lymphoma-associated antigens. AB - Diffuse large B-cell lymphoma (DLBCL) accounts for 30-40% of all adult non Hodgkin's lymphomas, yet the understanding of its underlying genetic abnormalities remains poor. Our present study used the serological analysis of recombinant cDNA expression libraries (SEREX) technique to identify DLBCL associated antigens. SEREX screening of testis libraries has previously identified cancer-testis antigens (CTAs) that may act as disease-specific targets for immunotherapy. Screening a testis cDNA expression library with serum from a DLBCL patient identified a total of 94 positive clones, representing 28 distinct antigens. Two of these antigens were novel, 8 were previously uncharacterised, and the remainder were proteins of known function. Screening of the antigens with sera from DLBCL (n = 10), acute myeloid leukaemia (AML, n = 10) and chronic myeloid leukaemia (CML, n = 10) patients, alongside normal healthy donor controls (n = 20), revealed that 7 of the antigens were recognised by DLBCL sera but not normal donor sera, whilst 2 of these antigens were also recognised by leukaemic sera. Some of the genes identified here were already known to be transcribed in DLBCL. The mRNA expression of the majority of the remaining antigens was confirmed in DLBCL cell lines using reverse-transcriptase PCR (RT-PCR). Our study identified a number of DLBCL associated antigens that may be suitable as prognostic/diagnostic markers and/or for the immunotherapy of haematologic malignancies. PMID- 15122590 TI - Gene expression analysis in Interleukin-12-induced suppression of mouse mammary carcinoma. AB - Interleukin-12 (IL-12) has potent antitumor activities via natural killer cells and cytotoxic T lymphocytes. However, the molecular mechanisms whereby IL-12 induces tumoricidal activities are poorly understood. Here, we report the genome wide analysis of gene expression in a primary murine mammary carcinoma model that resembles human breast cancer, following the therapeutic application of recombinant IL-12, which restricted tumor growth and metastasis. IL-12 was able to curtail neovascularization in the tumor as well as enhance the number of tumor infiltrating lymphocytes. Comprehensive examination of global gene expression revealed IL-12-induced molecular changes associated with tumor regression and reduced lung metastasis, thus providing a high-resolution snapshot of a host response against a developing malignancy and a rich source of potential targets for therapeutic intervention of breast cancer. PMID- 15122591 TI - Education and risk of breast cancer in the Norwegian-Swedish women's lifestyle and health cohort study. AB - A positive relationship between level of education and female breast cancer risk is well supported by scientific evidence, but few previous studies could adjust for all relevant potential confounding factors. The authors' purpose was to examine how risk for breast cancer varies with level of education and to identify factors that explain this variation, using data from a prospective cohort study including 102860 women from Norway and Sweden who responded to an extensive questionnaire in 1991/1992; 1090 incident primary invasive breast cancer cases were revealed during follow-up, which ended in December 1999. The Cox Proportional Hazards Model was used to calculate relative risks (RR) with 95% confidence intervals (CI). Women with more than 16 years of education had a 36% increased risk compared to the lowest educated (7-9 years) (Age adjusted RR=1.36, 95% CI: 1.10, 1.68). This relationship was slightly stronger among postmenopausal (RR 1.51) than among premenopausal (RR 1.25) women. In both groups, however, the relative risk estimates turned close to unity by adjustment for parity, age at first birth, body mass index (BMI), height, age at menarche, menopausal status, use of oral contraceptives and consumption of alcohol. The overall multivariate relative risk among the highest educated women was 1.04 (95% CI 0.82-1.32). The results of our study suggest a clear positive gradient in risk for breast cancer by level of education, which can be fully explained by established breast cancer risk factors. PMID- 15122592 TI - Serum insulin-like growth factor-I, insulin-like growth factor binding protein-3, and the risk of pancreatic cancer death. AB - Recent epidemiological studies have shown that high serum levels of insulin-like growth factor-I (IGF-I) are associated with an increased risk of lung, colon, breast and prostate cancer. Since very few studies have addressed the role of serum levels of IGF-I in the development of pancreatic cancer, we conducted a nested case-control study to examine this association. The analysis involved 69 case subjects who died from pancreatic cancer during the follow-up period of the study, and 207 control subjects matched for sex, age(+/-1 year) and study area, selected randomly from a cohort of 10364 individuals. Serum levels of IGF-I and IGF binding protein-3 (IGFBP-3) were measured by immunoradiometric assay, using commercially available kits. The odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using conditional logistic models. The levels of IGF-I were positively correlated with IGFBP-3 (r=0.55). There was a positive, but statistically insignificant association between serum levels of IGF-I and risk of death from pancreatic cancer, with subjects in the highest quartile having an OR of 2.31 (95% CI=0.70-2.64) compared to those in the lowest quartile. The risk of pancreatic cancer death increased significantly with increasing serum levels of IGFBP-3 (trend p=0.03). Further adjustment for IGFBP-3 or IGF-I slightly attenuated the positive associations. This nested case-control study showed that high serum levels of IGF-I and IGFBP-3 may be associated with an increased risk of death from pancreatic cancer. PMID- 15122593 TI - High fat and alcohol intakes are risk factors of postmenopausal breast cancer: a prospective study from the Malmo diet and cancer cohort. AB - Associations between intakes of relative fat, total alcohol and alcoholic beverages and risk of breast cancer were examined in a subsample of 11726 postmenopausal women from the MDC cohort. The MDC conducted baseline examinations from 1991 to 1996; the end of follow-up was 31 December 2001. Data were obtained by an interview-based diet history method, a structured questionnaire, anthropometric measurements and national and regional cancer registries. During 89602 person-years of follow-up, 342 incident cases were documented. Cox regression analysis examined breast cancer risks adjusted for potential confounders. Two energy-adjustment approaches (i.e., adjusting for total energy vs. adjusting for nonalcohol energy) were used. High total alcohol intake was associated with a nonsignificantly elevated risk. High wine intake was associated with a significantly elevated breast cancer risk (relative risk = 2.12, 95% CI 1.24-3.60). There were significant trends of increased breast cancer risk across quintiles of relative fat intake. Mutual adjustment did not affect risk estimates for total alcohol or relative fat intakes. The specific energy-adjustment approach did not influence associations differentially. PMID- 15122594 TI - GST, NAT, SULT1A1, CYP1B1 genetic polymorphisms, interactions with environmental exposures and bladder cancer risk in a high-risk population. AB - Tobacco smoking and occupation are major risk factors of bladder cancer via exposure to polycyclic aromatic hydrocarbons (PAHs) and aromatic amines. Glutathione S-transferase (GST) M1, T1 and P1 are involved in the detoxification of PAH reactive metabolites. Two N-acetyltransferase isozymes, NAT2 and NAT1, have major roles in catalyzing the N-acetylation and O-acetylation of aromatic amines. Cytochrome p450 1B1 (CYP1B1) and sulfotransferase 1A1 (SULT1A1) are also involved in the metabolism of PAHs and aromatic amines. It is hypothesized that the genetic polymorphisms of these metabolic enzymes have an effect on the individual susceptibility to bladder cancer in particular by interacting with relevant environmental exposures. A hospital-based case-control study among men in Brescia, Northern Italy recruited 201 incidence cases and 214 controls from 1997-2000. Occupational exposures were blindly coded by occupational physicians. Genotyping of polymorphisms were carried out with PCR-RFLP method. Unconditional multivariate logistic regression was applied to model the association between genetic polymorphisms and bladder cancer risk. Effect modifications by age of onset, smoking and occupational exposures to PAHs and aromatic amines were evaluated. We also conducted an analysis of interaction between genetic factors. GSTM1 and GSTT1 null genotype were associated with an increased risk of bladder cancer with an odds ratio (OR) of 1.69 (95% confidence interval [CI] = 1.11-2.56) and 1.74 (95% CI = 1.02-2.95), respectively. The effect of GSTM1 null was seen particularly in heavy smokers, and there was a combined effect with occupational exposure of aromatic amines (OR = 2.77, 95% CI = 1.08-7.10). We observed a trend (p-value < 0.01) of increasing cancer risk comparing subjects with normal GSTM1 and T1 activity to subjects with one (OR = 1.82, 95% CI = 1.16-2.85) or both null genotypes (OR = 2.58, 95% CI = 1.27-5.23). NAT2 slow acetylator was associated with marginally increased risk of bladder cancer (OR = 1.50, 95% CI = 0.99-2.27), and the OR for the joint effect with occupational exposure of aromatic amines was 3.26 (95% CI = 1.06-9.95). SULT1A1 Arg213His polymorphism showed a marginal protective effect. These findings suggest that individual susceptibility to bladder cancer may be modulated by GSTM1, GSTT1 and NAT2 polymorphisms. PMID- 15122595 TI - Passive smoking and breast cancer risk among non-smoking Chinese women. AB - Our purpose was to evaluate whether passive exposure to cigarette smoke may be related to breast cancer risk. Data from the Shanghai Breast Cancer Study, a large population-based study of 1459 breast cancer cases and 1556 controls aged 25-64 years, were analyzed. Respective response rates were 91.1% and 90.3%. Passive smoking questions were added to all face-to-face interviews 7 months into the study. Women were asked about exposure to their husbands' smoke at home as well as exposure in the workplace. Analyses were restricted to the 1013 cases and 1117 controls with passive tobacco smoke exposure data who had never actively smoked. Over 60% of controls reported some exposure to a husband's smoke and over 40% reported exposure to passive smoke in the workplace. Overall, there was no apparent association between any passive smoke exposure or exposure to a husband's smoke and breast cancer risk. There was some evidence of an elevated breast cancer risk associated with passive smoking exposure of 5 hr or more per day in the workplace (OR = 1.6, 95% confidence interval 1.0-2.4; p for trend = 0.02). This association warrants further investigation. PMID- 15122596 TI - Novel inhibitors of urokinase-type plasminogen activator and matrix metalloproteinase expression in metastatic cancer cell lines. AB - The plasminogen-activating (PA) and matrix metalloproteinase (MMP) enzyme systems are implicated in proteolytic turnover of the extracellular matrix (ECM) associated with biologic processes including wound healing, inflammation and angiogenesis. Aberrant expression of components of the PA and MMP enzyme systems occurs in the pathogenesis of metastatic cancer. Oxamflatin (Ox), a novel hydroxamic acid derivative, inhibits u-PA mRNA expression and proteolytic activity while simultaneously upregulating the expression of the natural inhibitor of u-PA, plasminogen activator inhibitor type 2 (PAI-2) in metastatic cancer cells. We have characterized the effects of Ox and a novel derivative, Metacept-1 (MCT-1), on PA and MMP-mediated proteolysis and invasion in several metastatic tumor lines. Both compounds are able to inhibit u-PA-, MMP-2- and MMP 9-mediated gene expression at low micromolar concentrations as well as u-PA- and MMP-mediated proteolysis as assessed by zymography, with MCT-1 being the more effective of the 2 agents in some assays. Cellular invasion assays correlate with gene expression and zymography experiments identifying both Ox and MCT-1 as able to inhibit invasion of metastatic cancer cell lines through matrigel at nanomolar concentrations, with MCT-1 more effective than Ox in 2 of the 3 cancer cell lines assessed. PMID- 15122597 TI - Polymorphisms in the one-carbon metabolic pathway, plasma folate levels and colorectal cancer in a prospective study. AB - One-carbon (e.g., folate) metabolism plays a pivotal role in the etiology of colorectal cancer (CRC). Cytosolic serine hydroxymethyltransferase (cSHMT), methylenetetrahydrofolate dehydrogenase (MTHFD1) and glutamate carboxypeptidase II (GCPII) are key genes involved in this pathway. Several new polymorphisms have been identified and there is evidence implicating their functionality. We examined whether polymorphisms in these genes, i.e., cSHMT L474F, MTHFD1 R653Q and GCPII H475Y, modify the risk of CRC in the prospective Physicians' Health Study. Among the 270 incident CRC cases and 453 controls, none of the one-carbon polymorphisms were associated with risk of CRC. Compared to the wild-type genotype, the multivariate-adjusted relative risks and 95% confidence intervals were 1.14 [0.68, 1.93] for cSHMT 474FF, 1.04 [0.67, 1.62] for MTHFD1 653QQ and 1.00 [0.55, 1.82] for GCPII 474HY. Furthermore, we examined the associations between one-carbon polymorphisms and folate status in terms of plasma folate and homocysteine levels in this population. No independent gene effect was observed. Although compound homozygous variants at cSHMT and MTHFD1 loci had the lowest plasma folate levels compared to other compound genotypes, no significant gene gene interactions were observed. Findings from our prospective investigation indicate that these newly identified polymorphisms in one-carbon metabolizing genes have limited functionality in modifying folate status and related CRC risk. PMID- 15122598 TI - Role of HTLV-1 proviral DNA load and clonality in the development of adult T-cell leukemia/lymphoma in asymptomatic carriers. PMID- 15122600 TI - Aortic dissection. PMID- 15122601 TI - Nosocomial infection. PMID- 15122602 TI - Techniques of reconstruction after total gastrectomy for cancer. AB - BACKGROUND: Jejunal pouch formation and restoration of duodenal transit have been suggested for reconstruction after total gastrectomy. Opinions about the clinical value vary. METHODS: The literature was searched for prospective randomized trials comparing reconstructive procedures after total gastrectomy for malignancy. Reports with at least an English or German abstract were included. Immediate results were evaluated in terms of postoperative deaths and complications. Long-term outcome was analysed using trends in bodyweight and quality of life. RESULTS: A total of 19 randomized trials including 866 patients was identified. The operative risk of total gastrectomy was low, with a median mortality rate of 0 (range 0-22) per cent, irrespective of the method of reconstruction. Neither gastric substitution nor restoration of duodenal transit was associated with significant procedure-related complications. Results for specific reconstructions varied considerably within and between individual trials. Jejunal pouch reconstruction, but not restoration of duodenal passage, was associated with improved food intake and a tendency for weight gain in the early postoperative months. A favourable perception of quality of life persisted in the long term in some studies. CONCLUSION: Preservation of duodenal transit offers little clinical benefit. Construction of a small-bowel reservoir after total gastrectomy should be considered to improve early postoperative eating capacity, bodyweight and quality of life. PMID- 15122603 TI - Neoadjuvant therapy for oesophagogastric cancer. AB - BACKGROUND: The prognosis after surgery for oesophagogastric cancer remains poor. METHODS: This review clarifies current indications for neoadjuvant therapy for oesophageal and gastric cancer. A systematic literature research and evaluation of data from international cancer meetings were carried out. RESULTS: Recently published results of large randomized phase III trials underscore the potential value of neoadjuvant treatment for oesophagogastric cancer. However, it remains uncertain which subgroups of patients should routinely undergo preoperative therapy. Metabolic response evaluation during neoadjuvant treatment is a promising tool for the selection of responding patients. CONCLUSION: Neoadjuvant chemotherapy is a valid option for locally advanced oesophageal and gastric cancer. In the future, more effective and better tolerated treatment strategies, tailored to the specific tumour characteristics of each individual, should be possible. PMID- 15122604 TI - Randomized clinical trial of laparoscopic versus open fundoplication for gastro oesophageal reflux. AB - BACKGROUND: The aim of this study was to compare the long-term results of laparoscopic and open antireflux surgery in a randomized clinical trial by investigating subjective and objective outcomes. METHODS: Sixty patients with gastro-oesophageal reflux disease (GORD) were randomized to laparoscopic or open 360 degrees fundoplication. Subjective evaluation using disease-specific and generic questionnaires and structured interviews, and objective evaluation by endoscopy, oesophageal manometry and 24-h pH monitoring, were performed before operation and 1 month, 6 months and 5 years after surgery. RESULTS: Two patients in the laparoscopic group had reoperations for hiatal stricture; one patient in the open group had repair of an incisional hernia, and one patient in each group had surgery for intestinal obstruction. There were no differences in the subjective outcomes of diet, sleep, medication, patient satisfaction and symptoms of GORD after 5 years. Nor were there any differences in objective outcomes determined by endoscopy, manometry or 24-h pH monitoring. Well-being was decreased in all patients before operation but was restored to normal or above normal values after fundoplication, regardless of the type of surgery. Seven of 28 patients in the open group had complaints regarding the scar. CONCLUSION: Elimination of GORD symptoms improved well-being and eliminated the need for daily acid suppression in most patients, no matter which procedure was employed. These results were apparent 1 month after the operation and were still valid 5 years later. PMID- 15122605 TI - Randomized clinical trial of distal anastomotic interposition vein cuff in infrainguinal polytetrafluoroethylene bypass grafting. AB - BACKGROUND: The aim was to examine the effect of a Miller vein cuff at the distal anastomosis on the medium- to long-term patency and limb salvage rates of femoral to above-knee and femoral to below-knee popliteal artery polytetrafluoroethylene (PTFE) bypasses. METHODS: This study involved extended follow-up of the original cohort of patients included in a previously reported multicentre randomized clinical study. Outcome measures were bypass graft patency and limb salvage. RESULTS: Two hundred and sixty-one bypass operations were originally randomized. For this study, full data were available on 235 (120 with a Miller cuff, 115 without). The cumulative 5-year patency rate for above-knee bypasses with a Miller cuff was 40 per cent, compared with 42 per cent for non-cuffed bypasses (P = 0.702). The cumulative 3-year patency rate for below-knee bypasses with a Miller cuff was 45 per cent, compared with 19 per cent for non-cuffed bypasses (P = 0.018). A Miller cuff had no significant effect on limb salvage for above-knee or below-knee bypasses. CONCLUSION: Three-year patency rates of femoral to below knee popliteal PTFE bypasses were improved by a Miller cuff. Miller cuffs had no effect on patency rates for femoral to above-knee popliteal bypasses at 5 years and did not improve limb salvage in either group. PMID- 15122606 TI - Outcome of abdominal aortic aneurysm repair in the era of endovascular treatment. AB - BACKGROUND: The effect on outcome of the introduction of endovascular techniques for the exclusion of abdominal aortic aneurysm (AAA) is largely unknown. The aim of the study was to contrast the early and mid-term outcome after open and endovascular AAA repair. METHODS: Consecutive patients who underwent transfemoral endovascular aneurysm repair (EVAR; n = 93) between April 1998 and January 2003, or conventional open aneurysm repair in the time intervals before (n = 113) and after (n = 82) the introduction of EVAR were studied. All data were collected prospectively. All patients underwent elective and primary repair. Patient survival was calculated by the Kaplan-Meier method and the possible predictive value of more than 25 perioperative variables on five outcome variables (length of intensive care unit stay, morbidity, and 30-day, 6-month and overall mortality) was assessed by multivariate analysis. RESULTS: There were no differences in risk profiles between the three groups. Selection for EVAR was based on anatomical grounds only. All five outcome variables were significantly better with EVAR than either former or recent open repair. The only significant predictors of failure were advanced age and the need to perform an additional procedure for former open repair; hypertension and the preoperative use of anticoagulants for recent open repair; and renal insufficiency and pulmonary insufficiency for EVAR. CONCLUSION: EVAR offered considerable benefits compared with conventional open repair at early and mid-term follow-up, which was not explained by selection of patients with a favourable risk profile. In the current era, in which patients are selected for open repair as a consequence of unfavourable anatomy, morbidity and mortality rates following conventional open treatment of AAA have increased at early and mid-term, but not at long-term, follow-up. PMID- 15122607 TI - Total thyroidectomy for clinically benign disease of the thyroid gland. AB - BACKGROUND: The role of total thyroidectomy in the treatment of patients with benign thyroid disease remains controversial. However, this procedure may be appropriate when both thyroid lobes are involved and when the risk of recurrence is significant. This study is a review of a 15-year experience of total thyroidectomy for benign disease. METHODS: Between 1988 and 2002, 834 patients underwent total thyroidectomy for clinically benign disease at the Sydney Head and Neck Cancer Institute, Royal Prince Alfred Hospital. There were 128 men and 706 women with a mean age of 52 (range 16-91) years. Indications for surgery were euthyroid multinodular goitre (MNG), toxic MNG and Graves' disease in 730 (87.5 per cent), 57 (6.8 per cent) and 47 (5.6 per cent) respectively. A total of 74 patients had previously undergone partial thyroidectomy. RESULTS: The incidence of temporary recurrent laryngeal nerve palsy was 2.3 per cent and that of temporary hypoparathyroidism 14.4 per cent. Permanent recurrent laryngeal nerve palsy occurred in 1.1 per cent, and 2.4 per cent of patients had permanent hypoparathyroidism. Neither the initial clinical diagnosis nor a history of previous treatment significantly influenced the rate of complications. The incidence of malignancy, other than incidental microscopic papillary carcinoma, was 4.6 per cent. CONCLUSION: Total thyroidectomy has an important role in the management of patients with benign disease when both lobes of the thyroid gland are involved. This approach avoids disease recurrence and the increased risk of morbidity associated with secondary operation. PMID- 15122608 TI - Infrared ray electronic endoscopy combined with indocyanine green injection for detection of sentinel nodes of patients with gastric cancer. AB - BACKGROUND: To avoid unnecessary lymphadenectomy in patients with cancer accurate diagnosis of the sentinel lymph node (SLN) is important. METHODS: This report examined the initial clinical use of infrared ray electronic endoscopy (IREE) combined with indocyanine green (ICG) injection for SLN detection in 84 patients with gastric cancer not invading the subserosa (75 T1 N0 M0 and nine T2 N0 M0 tumours, according to tumour node metastasis classification). RESULTS: There were no adverse events after injection of ICG. At least one SLN was detected in all but one patient by both ICG injection alone and by IREE with ICG. Eleven of the 84 patients had lymph node metastasis. SLNs detected by ICG injection alone did not include metastases in four of 11 patients, whereas IREE with ICG detected SLNs containing lymph node metastases in all 11 patients. Moreover, SLNs illuminated by IREE with ICG included all metastases among the 105 regional lymph nodes in the 11 patients; no metastatic lymph nodes were identified among 154 non SLNs. CONCLUSION: IREE combined with ICG injection may efficiently detect SLNs that contain metastases in patients with gastric cancer. PMID- 15122609 TI - Oesophageal and gastric bile exposure after gastroduodenal surgery with Henley's interposition or a Roux-en-Y loop. AB - BACKGROUND: The degree which the various reconstruction techniques prevent bile reflux after gastroduodenal surgery has been poorly studied. METHODS: Bile exposure in the intestinal tract just proximal to the jejunal loop was measured with the Bilitec 2000 device for 24 h after gastroduodenal surgery in three groups of patients. Group 1 comprised 24 patients with a 60-cm Henley's loop after total gastrectomy. Group 2 included 31 patients with a 60-cm Roux-en-Y loop after total (22 patients) or subtotal (nine) gastrectomy. Group 3 contained 21 patients with a 60-cm Roux-en-Y loop anastomosed to the proximal duodenum as part of a duodenal switch operation for pathological transpyloric duodenogastric reflux. Bile exposure, measured as the percentage time with bile absorbance greater than 0.25, was classified as nil, within the range of a control population of healthy subjects, or pathological (above the 95th percentile for the control population). Reflux symptoms were scored and all patients had upper gastrointestinal endoscopy. RESULTS: Bile was detected in the intestine proximal to the loop in none of 24 patients in group 1, eight of 31 in group 2 and 12 of 21 in group 3 (P < 0.001). The mean reflux symptom score increased with the degree of bile exposure, and the proportion of patients with oesophagitis or gastritis correlated well with the extent of bile exposure (P < 0.001). CONCLUSION: A long Henley's loop was more effective in preventing bile reflux than a long Roux-en-Y loop. Bilitec data correlated well with the severity of reflux symptoms and the presence of mucosal lesions. PMID- 15122610 TI - Curative resection is the single most important factor determining outcome in patients with pancreatic adenocarcinoma. AB - BACKGROUND: Mortality rates associated with pancreatic resection for cancer have steadily decreased with time, but improvements in long-term survival are less clear. This prospective study evaluated risk factors for survival after resection for pancreatic adenocarcinoma. METHODS: Data from 366 consecutive patients recorded prospectively between November 1993 and September 2001 were analysed using univariate and multivariate models. RESULTS: Fifty-eight patients (15.8 per cent) underwent surgical exploration only, 97 patients (26.5 per cent) underwent palliative bypass surgery and 211 patients (57.7 per cent) resection for pancreatic adenocarcinoma. Stage I disease was present in 9.0 per cent, stage II in 18.0 per cent, stage III in 68.7 per cent and stage IV in 4.3 per cent of patients who underwent resection. Resection was curative (R0) in 75.8 per cent of patients. Procedures included pylorus-preserving Whipple resection (41.2 per cent), classical Whipple resection (37.0 per cent), left pancreatic resection (13.7 per cent) and total pancreatectomy (8.1 per cent). The in-hospital mortality and cumulative morbidity rates were 2.8 and 44.1 per cent respectively. The overall actuarial 5-year survival rate was 19.8 per cent after resection. Survival was better after curative resection (R0) (24.2 per cent) and in lymph node negative patients (31.6 per cent). A Cox proportional hazards survival analysis indicated that curative resection was the most powerful independent predictor of long-term survival. CONCLUSION: Resection for pancreatic adenocarcinoma can be performed safely. The overall survival rate is determined by the radicality of resection. Patients deemed fit for surgery who have no radiological signs of distant metastasis should undergo surgical exploration. Resection should follow if there is a reasonable likelihood that an R0 resection can be obtained. PMID- 15122611 TI - Use of the time-signal intensity curve from dynamic magnetic resonance imaging to evaluate remnant pancreatic fibrosis after pancreaticojejunostomy in patients undergoing pancreaticoduodenectomy. AB - BACKGROUND: This study assessed the value of the time-signal intensity curve (TIC) obtained from dynamic magnetic resonance imaging (MRI) in the evaluation of remnant pancreatic fibrosis after pancreaticojejunostomy in patients undergoing pancreaticoduodenectomy. METHODS: Two modes of pancreaticojejunostomy-duct-to mucosa anastomosis (DMA; 24 patients) and pancreatojejunoserosal anastomosis (PJSA; 22 patients)-were used in 46 consecutive patients undergoing pancreaticoduodenectomy. All patients underwent dynamic contrast-enhanced MRI of the pancreas before pancreaticoduodenectomy. Retrospective review of the pancreatic magnetic resonance images and histological examination of the pancreas were performed, and the patterns of TICs from dynamic MRI were compared with the degree of pancreatic fibrosis. Dynamic MRI of the residual pancreas was carried out for 1-3 years after pancreaticoduodenectomy in 26 patients (14 DMA, 12 PJSA) who had a histologically verified normal pancreas with no fibrosis at the time of pancreaticoduodenectomy. RESULTS: Evaluation of preoperative dynamic magnetic resonance images showed that a pancreatic TIC with a rapid rise to a peak followed by a rapid decline (type I) was characteristic of a normal pancreas without fibrosis. Pancreatic TICs with a slow rise to a peak followed by a slow decline or a plateau (types II and III) indicated a fibrotic pancreas. Postoperative pancreatic MRI demonstrated that six of 12 patients with a PJSA had a type II TIC, whereas 12 of 14 patients with a DMA had a type I curve (P = 0.046). CONCLUSION: The TIC obtained from dynamic MRI is a reliable indicator of fibrosis in the remnant pancreas after pancreaticoduodenectomy. Use of a DMA was associated with a lower risk of pancreatic fibrosis 1-3 years after surgery than a PJSA. PMID- 15122612 TI - Timing of urgent laparoscopic cholecystectomy does not influence conversion rate. AB - BACKGROUND: The optimal treatment of acute gallstone disease is urgent laparoscopic cholecystectomy, but there is confusion about the effect of delay in operation on conversion rates. Most reports suggest that delay beyond 3 or 4 days leads to a higher conversion rate. This study assessed the conversion rate in relation to the timing of laparoscopic surgery. METHODS: This institution operates a specialist-led protocol for the urgent management of all admissions with acute gallstone disease. Data were collected prospectively over 6 months. RESULTS: Between March and August 2002, 84 patients with acute gallstone disease underwent urgent laparoscopic cholecystectomy at the index admission with an overall conversion rate of 12 per cent. Four of 40 procedures carried out within 3 days of admission were converted, compared with six of 44 after 3 days. Five of 46 carried out within 4 days of admission were converted, compared with five of 38 after 4 days. There were no deaths and one common bile duct injury. CONCLUSION: As long as the procedure is carried out by experienced upper gastrointestinal surgeons working within a specialist-led protocol, the conversion rate for laparoscopic cholecystectomy can be as low as 12 per cent. The timing of urgent laparoscopic cholecystectomy has no impact on the conversion rate. PMID- 15122613 TI - Emergency presentation of colorectal cancer is associated with poor 5-year survival. AB - BACKGROUND: Previous studies have reported that emergency presentation of colorectal cancer is associated with poor outcome. Many of these studies were small and most were not adjusted for case mix. The aim of this study was to establish, after adjusting for case mix, the magnitude of the differences in postoperative mortality and survival between patients undergoing elective surgery and those presenting as an emergency. METHODS: Three thousand two hundred patients who underwent surgery for colorectal cancer between 1991 and 1994 in Scotland were studied. Five-year survival rates and adjusted hazard ratios were calculated. RESULTS: Some 1603 (72.4 per cent) of 2214 elective patients had a potentially curative resection compared with 632 (64.1 per cent) of 986 patients who presented as an emergency (P < 0.001). Following curative resection, the postoperative mortality rate was 2.8 per cent after elective and 8.2 per cent after emergency operation (P < 0.001). Overall survival at 5 years was 57.5 per cent after elective and 39.1 per cent after emergency curative surgery (P < 0.001); cancer-specific survival at 5 years was 70.9 and 52.9 per cent respectively (P < 0.001). The adjusted hazard ratio for overall survival after emergency relative to elective surgery was 1.68 (95 per cent confidence interval (c.i.) 1.49 to 1.90; P < 0.001) and that for cancer-specific survival was 1.90 (95 per cent c.i. 1.62 to 2.22; P < 0.001). CONCLUSION: Following apparently curative resection for colorectal cancer, there was an excess of both cancer related and intercurrent deaths in patients who presented as an emergency. PMID- 15122614 TI - Influence of volume and specialization on survival following surgery for colorectal cancer. AB - BACKGROUND: Previous studies have shown that significant surgeon-related differences in survival exist following surgery for colorectal cancer. It is not clear whether these differences were due to differences in caseload or degree of specialization. METHODS: Outcome in 3200 patients who underwent resection for colorectal cancer between 1991 and 1994 was analysed on the basis of caseload and degree of specialization of individual surgeons. Five-year survival rates, and the corresponding hazard ratios adjusted for case mix, were calculated. RESULTS: Cancer-specific survival rate at 5 years following curative resection varied among surgeons from 53.4 to 84.6 per cent; the adjusted hazard ratios varied from 0.48 to 1.55. Cancer-specific survival rate at 5 years following curative resection was 70.2, 62.0 and 65.9 per cent for surgeons with a high, medium and low case volume respectively. There were no consistent differences in the adjusted hazard ratios by volume. Cancer-specific survival rate at 5 years following curative resection was 72.7 per cent for those treated by specialists and 63.8 per cent for those treated by non-specialists; the adjusted hazard ratio for non-specialists was 1.35 (95 per cent confidence interval 1.13 to 1.62; P = 0.001). CONCLUSION: The differences in outcome following apparently curative resection for colorectal cancer among surgeons appear to reflect the degree of specialization rather than case volume. It is likely that increased specialization will lead to further improvements in survival. PMID- 15122615 TI - Working capacity and resting energy expenditure after ileal pouch-anal anastomosis. AB - BACKGROUND: The aim was to evaluate the working capacity and resting energy expenditure in patients who had undergone restorative proctocolectomy. METHODS: Of 72 patients operated on between April 1990 to September 1998, 51 were eligible and 38 participated in the study. Resting energy was assessed by indirect calorimetry, and working capacity by ergospirometry on an exercise bicycle. RESULTS: The median functional score was 2 (range 0-7). Oxygen uptake during rest was reduced for men compared with predicted values. The corresponding values for women were in keeping with predicted values. The median working capacity was 96 (range 59-102) per cent for women and 91 (range 51-113) per cent for men, compared with reference values of maximum workload based on age, height and sex. There was no correlation between functional score and any other variable measured. CONCLUSION: Patients who have undergone restorative proctocolectomy for ulcerative colitis have normal resting energy expenditure and working capacity. PMID- 15122616 TI - Fistuloclysis can successfully replace parenteral feeding in the nutritional support of patients with enterocutaneous fistula. AB - BACKGROUND: Use of total parenteral nutrition (TPN) in patients with acute intestinal failure due to enteric fistulation might be avoided if a simpler means of nutritional support was available. The aim of this study was to determine whether feeding via an intestinal fistula (fistuloclysis) would obviate the need for TPN. METHODS: Fistuloclysis was attempted in 12 patients with jejunocutaneous or ileocutaneous fistulas with mucocutaneous continuity. Feeding was achieved by inserting a gastrostomy feeding tube into the intestine distal to the fistula. Infusion of enteral feed was increased in a stepwise manner, without reinfusion of chyme, until predicted nutritional requirements could be met by a combination of fistuloclysis and regular diet, following which TPN was withdrawn. Energy requirements and nutritional status were assessed before starting fistuloclysis and at the time of reconstructive surgery. RESULTS: Fistuloclysis replaced TPN entirely in 11 of 12 patients. Nutritional status was maintained for a median of 155 (range 19-422) days until reconstructive surgery could be safely undertaken in nine patients. Two patients who did not undergo surgery remained nutritionally stable over at least 9 months. TPN had to be recommenced in one patient. There were no complications associated with fistuloclysis. CONCLUSION: Fistuloclysis appears to provide effective nutritional support in selected patients with enterocutaneous fistula. PMID- 15122617 TI - Delayed portal vein thrombosis after experimental radiofrequency ablation near the main portal vein. AB - BACKGROUND: Portal venous blood flow may protect adjacent tumour cells from thermal destruction with radiofrequency ablation (RFA). This study aimed to investigate the local effect of RFA on the main portal vein branch, and the completeness of cellular ablation in its vicinity, with or without a Pringle manoeuvre using a porcine model. METHODS: This was an in vivo study on 23 domestic pigs. RFA using a cooled-tip electrode was performed 5 mm from the left main portal vein branch under ultrasonographic guidance for 12 min with (n = 10) or without (n = 10) a Pringle manoeuvre. Ten pigs were killed 4 h after the procedure to study the early effects of RFA and ten others were killed 1 week later to determine any delayed effect. As a control, sham operations with a Pringle manoeuvre for 12 min were performed on three pigs. The flow velocity changes of portal vein and hepatic artery were measured using Doppler ultrasonography, and the completeness of cellular ablation around the portal vein was assessed qualitatively by histochemical staining and quantitatively by measuring intracellular levels of adenosine 5'-triphosphate (ATP). RESULTS: In the absence of the Pringle manoeuvre, there was no significant change in mean(s.d.) portal vein flow velocity before RFA (20.0(3.5) cm/s) and at 4 h (18.5(2.5) cm/s) (P = 0.210) and 1 week (19.5(2.2) cm/s) (P = 0.500) after the procedure. Gross and histological examination of the portal vein branches showed no damage without the Pringle manoeuvre. In all pigs that underwent RFA with a Pringle manoeuvre, the portal vein was occluded 1 week after the operation; histological examination of the affected portal vein showed severe thermal injury and associated venous thrombosis. The local effect of RFA on the hepatic artery was similar. With intact portal blood flow during RFA, complete ablation of liver tissue around the pedicle was demonstrated by histochemical staining and measurement of the intracellular ATP concentration. CONCLUSION: RFA was safe when applied close to the main portal vein branch without a Pringle manoeuvre, with complete cellular destruction. Use of the Pringle manoeuvre resulted in delayed portal vein and hepatic artery thrombosis and injury to the hepatic artery and bile duct. PMID- 15122618 TI - Incidence and outcome of surgical repair of spigelian hernia. AB - BACKGROUND: A spigelian hernia develops through a defect present in a locus minoris resistentiae between abdominal muscles. It is generally considered rare but is probably underdiagnosed. This study was undertaken to characterize the history, presenting features and efficacy of surgical correction of spigelian hernia. METHODS: Medical and surgical charts of 25 patients treated for a spigelian hernia over 22 years were studied retrospectively. Patients were invited for an outpatient interview and physical examination. RESULTS: Dominant symptoms were an intermittent palpable mass (22 patients) and postural pain (16). A quarter of the patients reported a history of other hernias. Two individuals presented with a painful palpable mass and signs of bowel obstruction necessitating emergency small bowel resection. Hernia repair was performed by primary closure in 20 patients and by use of mesh graft in five. During a mean follow-up of 6.1 years, one early recurrence was observed. Two patients reported mild tenderness in the operated area but were not impaired in daily activities. CONCLUSION: Spigelian hernia is commonly encountered and requires surgical treatment because of the risk of strangulation. Operative treatment is simple and effective in the long term. PMID- 15122620 TI - Preoperative probability model for predicting overall survival after resection of pulmonary metastases from colorectal cancer (Br J Surg 2004; 91: 112-120). PMID- 15122621 TI - Preoperative probability model for predicting overall survival after resection of pulmonary metastases from colorectal cancer (Br J Surg 2004; 91: 112-120). PMID- 15122622 TI - Increased vascular endothelial growth factor production in fibroblasts isolated from strictures in patients with Crohn's disease (Br J Surg 2004; 91: 72-77). PMID- 15122625 TI - Surgical site infection after groin hernia repair (Br J Surg 2004; 91: 105-111). PMID- 15122626 TI - Meta-analysis of randomized clinical trials comparing open and laparoscopic inguinal hernia repair (Br J Surg 2004; 91: 253. PMID- 15122630 TI - The unique role of fluorine in the design of active ingredients for modern crop protection. AB - The task of inventing and developing active ingredients with useful biological activities requires a search for novel chemical substructures. This process may trigger the discovery of whole classes of chemicals of potential commercial interest. Similar biological effects can often be achieved by completely different compounds. However, compounds within a given structural family may exhibit quite different biological activities depending on their interactions with different intracellular proteins like enzymes or receptors. By varying the functional groups and structural elements of a lead compound, its interaction with the active site of the target protein, as well as its physicochemical, pharmacokinetic, and dynamic properties can be improved. In this context, the introduction of fluorine into active ingredients has become an important concept in the quest for a modern crop protection product with optimal efficacy, environmental safety, user friendliness, and economic viability. Fluorinated organic compounds represent an important and growing family of commercial agrochemicals. A number of recently developed agrochemical candidates represent novel classes of chemical compounds with new modes of action; several of these compounds contain new fluorinated substituents. However, the complex structure activity relationships associated with biologically active molecules mean that the introduction of fluorine can lead to either an increase or a decrease in the efficacy of a compound depending on its changed mode of action, physicochemical properties, target interaction, or metabolic susceptibility and transformation. Therefore, it is still difficult to predict the sites in a molecule at which fluorine substitution will result in optimal desired effects. PMID- 15122631 TI - The influence of fluorinated molecules (semiochemicals and enzyme substrate analogues) on the insect communication system. AB - Can the introduction of fluorine atoms affect the bioactivity of natural semiochemicals? Can fluorine contribute in the creation of specific enzyme inhibitors to interrupt or disrupt the insect communication system? The first step for the bioactivity of a molecule is interaction with the biological sensor. Hydrogen and fluorine are almost bioisosteric and the receptor site of the enzyme can still recognize and accept the fluoro analogue of its natural substrate. However, the peculiar electronegativity of the fluorine atom can affect the binding, absorption, and transport of the molecule. The differences in the molecule's electronic properties can lead to differences in the chemical interactions between the receptor and the fluorinated substrate. Fluorine introduction can modify the metabolic stability and pathway of the semiochemicals in many different ways. Fluorinated analogues can show synergism, inhibition, or hyperagonism effects on insect behaviors, that is, the activity of the nonfluorinated parent compounds can be mimicked, lost, or increased. In any case, the fluorinated molecules can interact with the bioreceptors in a new and disrupting way. The semiochemicals are olfactory substances: fluorine can affect their volatility or smell. Production of semiochemicals from exogenous substances, perception at antennal receptors, and processing of biological responses are the main steps of communication among insects. In the production step, the fluorinated molecules can interact with enzymes that catalyze the biosynthesis of the natural pheromones. In the perception step, fluorinated semiochemicals can interact with the olfactory receptor cells; this often leads to totally unpredictable behaviors. Fluorinated molecules have been developed as probes to elucidate the complex chemorecognition processes of insects. Many of these molecules have been tested to find highly effective behavior-modifying chemicals. New analogues have been synthesized to investigate the metabolic pathway of a pheromone molecule and many of them are promising disrupting agents. Despite such titanic research efforts, the results have often been random, rational trends in the induced behaviors have sometimes been impossible to find, and practical applications of the fluorinated semiochemicals are still uncertain. PMID- 15122632 TI - Organic fluorine: odd man out. PMID- 15122633 TI - The polar hydrophobicity of fluorinated compounds. PMID- 15122634 TI - Use of fluorine in the medicinal chemistry and chemical biology of bioactive compounds--a case study on fluorinated taxane anticancer agents. PMID- 15122635 TI - Fluorine in medicinal chemistry. AB - Fluorinated compounds are synthesized in pharmaceutical research on a routine basis and many marketed compounds contain fluorine. The present review summarizes some of the most frequently employed strategies for using fluorine substituents in medicinal chemistry. Quite often, fluorine is introduced to improve the metabolic stability by blocking metabolically labile sites. However, fluorine can also be used to modulate the physicochemical properties, such as lipophilicity or basicity. It may exert a substantial effect on the conformation of a molecule. Increasingly, fluorine is used to enhance the binding affinity to the target protein. Recent 3D-structure determinations of protein complexes with bound fluorinated ligands have led to an improved understanding of the nonbonding protein-ligand interactions that involve fluorine. PMID- 15122636 TI - Atropisomerism, biphenyls, and fluorine: a comparison of rotational barriers and twist angles. PMID- 15122637 TI - The coordination chemistry of fluorine in fluorocarbons. PMID- 15122638 TI - Experimental measurement of noncovalent interactions between halogens and aromatic rings. AB - Chemical double mutant cycles have been used to quantify the interactions of halogens with the faces of aromatic rings in chloroform. The halogens are forced over the face of an aromatic ring by an array of hydrogen-bonding interactions that lock the complexes in a single, well-defined conformation. These interactions can also be engineered into the crystal structures of simpler model compounds, but experiments in solution show that the halogen-aromatic interactions observed in the solid state are all unfavourable, regardless of whether the aromatic rings contain electron-withdrawing or electron-donating substituents. The halogen-aromatic interactions are repulsive by 1-3 kJ mol(-1). The interactions with fluorine are slightly less favourable than with chlorine and bromine. PMID- 15122639 TI - Fluorine interactions at the thrombin active site: protein backbone fragments H C(alpha)-C=O comprise a favorable C-F environment and interactions of C-F with electrophiles. AB - In a systematic fluorine scan of a rigid inhibitor to map the fluorophilicity/fluorophobicity of the active site in thrombin, one or more F substituents were introduced into the benzyl ring reaching into the D pocket. The 4-fluorobenzyl inhibitor showed a five to tenfold higher affinity than ligands with other fluorination patterns. X-ray crystal-structure analysis of the protein ligand complex revealed favorable C-F...H-C(alpha)-C=O and C-F...C=O interactions of the 4-F substituent of the inhibitor with the backbone H-C(alpha)-C=O unit of Asn98. The importance of these interactions was further corroborated by the analysis of small-molecule X-ray crystal-structure searches in the Protein Data Base (PDB) and the Cambridge Structural Database (CSD). In the C--F...C=O interactions that are observed for both aromatic and aliphatic C-F units and a variety of carbonyl and carboxyl derivatives, the F atom approaches the C=O C atom preferentially along the pseudotrigonal axis of the carbonyl system. Similar orientational preferences are also seen in the dipolar interactions C--F.C[triple chemical bond]N, C-F.C-F, and C-F...NO(2), in which the F atoms interact at sub van der Waals distances with the electrophilic centers. PMID- 15122640 TI - Halogenation of drugs enhances membrane binding and permeation. AB - Halogenation of drugs is commonly used to enhance membrane binding and permeation. We quantify the effect of replacing a hydrogen residue by a chlorine or a trifluoromethyl residue in position C-2 of promazine, perazine, and perphenazine analogues. Moreover, we investigate the influence of the position (C 6 and C-7) of residue CF(3) in benzopyranols. The twelve drugs are characterized by surface activity measurements, which yield the cross-sectional area, the air water partition coefficient, and the critical micelle concentration. By using the first two parameters (A(D) and K(aw)) and the appropriate membrane packing density, the lipid-water partition coefficients, are calculated in excellent agreement with the lipid-water partition coefficients measured by means of isothermal titration calorimetry for small unilamellar vesicles of 1-palmitoyl-2 oleoyl-sn-glycero-3-phosphocholine. Replacement of a hydrogen residue by a chlorine and a trifluoromethyl residue enhances the free energy of partitioning into the lipid membrane, on average by deltaG(lw) approximately -1.3 or -4.5 kJ mol(-1), respectively, and the permeability coefficient by a factor of approximately 2 or approximately 9, respectively. Despite exhibiting practically identical hydrophobicities, the two benzopyranol analogues differ in their permeability coefficients by almost an order of magnitude; this is due to their different cross-sectional areas at the air-water and lipid-water interfaces. PMID- 15122641 TI - Enzymatic fluorination in Streptomyces cattleya takes place with an inversion of configuration consistent with an SN2 reaction mechanism. AB - The stereochemical course of the recently isolated fluorination enzyme from Streptomyces cattleya has been evaluated. The enzyme mediates a reaction between the fluoride ion and S- adenosyl-L-methionine (SAM) to generate 5'-fluoro-5' deoxyadenosine (5'-FDA). Preparation of (5'R)-[5-(2)H(1)]-ATP generated (5'R)-[5 (2)H(1)]-5'-FDA in a coupled enzyme assay involving SAM synthase and the fluorinase. The stereochemical analysis of the product relied on (2)H NMR analysis in a chiral liquid-crystalline medium. It is concluded that the enzyme catalyses the fluorination with an inversion of configuration consistent with an S(N)2 reaction mechanism. PMID- 15122642 TI - Probing the proteolytic stability of beta-peptides containing alpha-fluoro- and alpha-hydroxy-beta-amino acids. AB - One of the benefits of beta-peptides as potential candidates for biological applications is their stability against common peptidases. Attempts have been made to rationalize this stability by altering the electron availability of a given amide carbonyl bond through the introduction of polar substituents at the alpha-position of a single beta-amino acid. Such beta-amino acids (beta homoglycine, beta-homoalanine), containing one or two fluorine atoms or a hydroxy group in the alpha-position, were prepared in enantiopure form. A versatile method for preparing these alpha-fluoro-beta-amino acids by the homologation of appropriate alpha-amino acids and C-OH->C-F or C=O-->CF(2) substitution with DAST, is described. Consequently, a series of beta-peptides possessing an electronically modified residue at the N terminus or embedded within the chain was synthesized, and their proteolytic stability was investigated against a selection of enzymes. All ten beta-peptides tested were resilient to proteolysis. Introducing a polar, sterically undemanding group, into the alpha-position of beta-amino acids in a beta-peptide chain does not appear to facilitate localized or general enzymatic degradation. PMID- 15122643 TI - Synthesis of 2'-aminoalkyl-substituted fluorinated nucleobases and their influence on the kinetic properties of hammerhead ribozymes. AB - Hammerhead ribozymes are ribonucleic acids that catalyse the hydrolytic cleavage of RNA. They interfere with gene expression in a highly specific manner and recognize the mRNA target through Watson-Crick base pairing. To overcome the problem of point mutations (Watson-Crick "mismatches") occurring in viral genomes, we developed 2'-aminoethyl-substituted fluorinated nucleosides, which are universal nucleobases. The highly efficient synthetic pathway, which features a direct phthaloylamination of a primary alcohol under Mitsunobu conditions, leads to modified phosphoroamidites. The 1'-deoxy-1'-(4,6-difluoro-1H benzimidazol-1-yl)-2'-(beta-aminoethyl)-beta-D-ribofuranose nucleoside analogue does not differentiate between the four natural nucleosides and leads to a RNA duplex that is as stable as the unmodified parent duplex. Upon incorporation into a ribozyme, the analogue's catalytic activity is equal for all four possible substrates, and the cleavage rates for the modified ribozymes are significantly higher (up to a factor of 13) than for the natural Watson-Crick "mismatch" base pairs. In agreement with the thermodynamic data obtained by measurement of the T(m) values of the RNA 12-mers, the cleavage rates for the 2'-substituted fluorinated benzimidazole derivative 4 are slightly higher than for the corresponding fluorinated benzene derivative 3. PMID- 15122644 TI - Evaluation of the molecular interactions of fluorinated amino acids with native polypeptides. PMID- 15122645 TI - Crystallographic evidence for isomeric chromophores in 3-fluorotyrosyl-green fluorescent protein. PMID- 15122647 TI - Characterization of the actions of AvTx 7 isolated from Agelaia vicina (Hymenoptera: Vespidae) wasp venom on synaptosomal glutamate uptake and release. AB - It has previously been shown that the denatured crude extract of Agelaia vicina wasp venom inhibits glutamate and GABA uptake in rat cerebral cortex synaptosomes. To identify the components responsible for these effects, the neurotoxin AvTx 7 (molecular weight of 1210 Da) was isolated from A. vicina venom and its effects on glutamate neurotransmission investigated. AvTx 7 inhibits glutamate uptake in a dose-dependent and uncompetitive manner. AvTx 7 was found to stimulate the glutamate release in the presence of calcium and sodium channel blockers, suggesting that its action is not mediated through these channels. AvTx 7 potentiates glutamate release in the presence of K(+) channel blockers tetraethylammonium and 4-aminopyridine, indicating that the toxin may act through these drugs-sensible K(+) channels. We suggest that AvTx 7 can be a valuable tool to enhance our understanding of K(+) channels' involvement in the release of glutamate. PMID- 15122648 TI - Nitric oxide and oxidative stress in brain and heart of normal rats treated with doxorubicin: role of aminoguanidine. AB - Doxorubicin (DOX) is a potent antitumor antibiotic drug known to cause severe cardiac toxicity. Moreover, its adverse effects were found to be extended to the cerebral tissue. Several mechanisms for this toxicity have been ascribed. Currently, one of the most accepted mechanisms is through free radicals; however, the exact role of nitric oxide (NO) is still unclear. Accordingly, a NO-synthase inhibitor with some antioxidant property, aminoguanidine (AG), was selected to examine its potential protective effect against DOX-induced toxicity. Male Wistar albino rats (150-200 g) were allocated into a normal control group, DOX-induced toxicity group, and DOX + AG-treated group. DOX was injected i.p. at a dose of 10 mg/kg divided into four equal injections over a period of 2 weeks. AG was injected i.p. at a dose of 100 mg/kg 1 h before each DOX injection. The animals were sacrificed 24 h after the last DOX injection and the following parameters were measured: serum lactate dehydrogenase (LDH) and creatine phosphokinase (CPK) activities, cardiac and cerebral contents of malondialdehyde (MDA), conjugated diene (CD), glutathione (GSH), NO, and cytosolic calcium, as well as superoxide dismutase (SOD) and glutathione peroxidase (GSHP(X)) activities. Cardiotoxicity was manifested by a marked increase in serum LDH and CPK in addition to the sharp increase in MDA reaching eightfolds the basal level. This was accompanied by significant increase in CD, NO, cytosolic calcium, SOD, and GSHP(X) content/activity by 69, 85, 76, 125, and 41% respectively as compared to normal control. On the other hand, GSH was significantly depressed. In brain, only significant increase in MDA and GSHP(X) and decrease in GSH were obtained but to a lesser extent than the cardiac tissue. AG treatment failed to prevent the excessive release of cardiac enzymes; however, it alleviated the adverse effects of DOX in heart. AG administration resulted in marked decrease in the elevated levels of MDA, NO, SOD, and GSHP(X), however, MDA level was still pathological. The altered parameters in brain were restored by AG. It is concluded that, AG could not provide complete protection against DOX-induced toxicity. Therefore, it is recommended that, maintenance of the endogenous antioxidant, GSH, and regulation of calcium homeostasis must be considered, rather than NO formation, to guard against DOX-induced toxicity. PMID- 15122649 TI - Cardiotoxicity of doxorubicin/paclitaxel combination in rats: effect of sequence and timing of administration. AB - The higher incidence of cardiotoxicity of doxorubicin (DOX)/paclitaxel (PTX) combination compared with DOX alone remains to be a major obstacle against effective chemotherapeutic treatment. We investigated the effect of sequence and time interval between administration of both drugs on the severity of cardiotoxicity of the combination. Male Wistar rats were divided into seven groups. DOX was administered intraperitoneally (i.p.) at a single dose of 5 mg x kg(-1) every other 2 days, 2 doses per week for a total cumulative dose of 20 mg x kg(-1). PTX was administered by an i.p. route at a dose of 20 mg x kg(-1) every other 2 days. Both drugs were injected either alone or sequentially in combination. In one case, DOX preceded PTX by 30 min and 24 h and in the other case, PTX preceded DOX by 30 min and 24 h. Cardiotoxicity was evaluated by both biochemical and histopathological examination, 48 h after the last DOX dose. DOX induced cardiotoxicity was manifested by abnormal biochemical changes including marked increases in serum creatine phosphokinase isoenzyme (CK-MB), lactate dehydrogenase (LDH), glutathione peroxidase (GSH-Px), and aspartate aminotransferase (AST) activity levels. Myocardial tissue from DOX-treated rats showed significant increases in malondialdehyde (MDA) production and total nitrate/nitrite (NOx) levels, parallel with depletion of "endogenous antioxidant reserve," including GSH contents and GSH-Px activity level. PTX treatment produced significant changes in the biochemical parameters measured by a lower magnitude than those changes produced by DOX alone. Combination of both drugs resulted in aggravation of DOX-induced cardiotoxicity regardless the sequence and time interval between administration of either drug. Administration of PTX 30 min and 24 h after DOX treatment showed exaggeration of combination-induced cardiotoxicity compared with the reverse sequence. This exacerbation was manifested by much more pronounced changes in serum and cardiac tissue parameters measured. Histopathological examination of ventricles of rat's heart revealed that DOX treatment produced myo-cytolysis and myocardial necrosis. Administration of PTX following DOX treatment showed extensive myocardial necrosis compared with those rats treated with either DOX alone or the reverse sequence of administration. Moreover, rats treated with PTX 24 h after DOX treatment showed exaggeration of the combination-induced cardiotoxicity. In conclusion, PTX might synergistically aggravate DOX-induced cardiotoxicity. The effect might be much more pronounced with those rats treated with PTX 24 h after DOX treatment. PMID- 15122650 TI - Carbon monoxide and oxidative stress in Desulfovibrio desulfuricans B-1388. AB - It has been shown that carbon monoxide (CO) in low concentration may be an active biochemical and physiological regulator of cell function. The bases of CO toxicity and cell protection are not clearly understood. To provide insights into these mechanisms, we measured superoxide production by D. desulfuricans B-1388 incubated anaerobically in Postgate medium with or without 5% CO. D. desulfuricans B-1388 growing with CO in the gas phase produced more superoxide radicals then control cells growing in Ar. When the cells were pregrown with CO, NADH oxidase and peroxidase activities were increased. The increase in peroxidase activities of cells growing under CO (particularly NADH peroxidase) suggested that H(2)O(2) was accumulated in cells. Superoxide dismutase (SOD) activity of cells decreased in exponential growth phase and increased in stationary phase. This may be due to CO concentration fall during CO oxidation by CO dehydrogenase. Altogether, our data suggest that superoxide production is a possible mechanism of CO toxicity. PMID- 15122651 TI - Benzene metabolism in human lung cell lines BEAS-2B and A549 and cells overexpressing CYP2F1. AB - Benzene is an occupational and environmental toxicant. The main human health concern associated with benzene exposure is leukemia. The toxic effects of benzene are dependent on its metabolism by the cytochrome p450 enzyme system. The cytochrome p450 enzymes CYP2E1 and CYP2F2 are the major contributors to the bioactivation of benzene in rats and mice. Although benzene metabolism has been shown to occur with mouse and human lung microsomal preparations, little is known about the ability of human CYP2F to metabolize benzene or the lung cell types that might activate this toxicant. Our studies compared bronchiolar derived (BEAS 2B) and alveolar derived (A549) human cell lines for benzene metabolizing ability by evaluating the roles of CYP2E1 and CYP2F1. BEAS-2B cells that overexpressed CYP2F1 and recombinant CYP2F1 were also evaluated. BEAS-2B cells overexpressing the enzyme CYP2F1 produced 47.4 +/- 14.7 pmols hydroxylated metabolite/10(6) cells/45 min. The use of the CYP2E1-selective inhibitor diethyldithiocarbamate and the CYP2F2-selective inhibitor 5-phenyl-1-pentyne demonstrated that both CYP2E1 and CYP2F1 are important in benzene metabolism in the BEAS-2B and A549 human lung cell lines. The recombinant expressed human CYP2F1 enzyme had a K(m) value of 3.83 microM and a V(max) value of 0.01 pmol/pmol p450 enzyme/min demonstrating a reasonably efficient catalysis of benzene metabolism (V(max)/K(m) = 2.6). Thus, these studies have demonstrated in human lung cell lines that benzene is bioactivated by two lung-expressed p450 enzymes. PMID- 15122652 TI - Characterization of triacsin C inhibition of short-, medium-, and long-chain fatty acid: CoA ligases of human liver. AB - Short-, medium-, and long-chain fatty acid:CoA ligases from human liver were tested for their sensitivity to inhibition by triacsin C. The short-chain fatty acid:CoA ligase was inhibited less than 10% by concentrations of triacsin C as high as 80 microM. The two mitochondrial xenobiotic/medium-chain fatty acid:CoA ligases (XM-ligases), HXM-A and HXM-B, were partially inhibited by triacsin C, and the inhibitions were characterized by low affinity for triacsin C (K(I) values > 100 microM). These inhibitions were found to be the result of triacsin C competing with medium-chain fatty acid for binding at the active site. The microsomal and mitochondrial forms of long-chain fatty acid:CoA ligase (also termed long-chain fatty acyl-CoA synthetase, or long-chain acyl-CoA synthetase LACS) were potently inhibited by triacsin C, and the inhibition had identical characteristics for both LACS forms. Dixon plots of this inhibition were biphasic. There is a high-affinity site with a K(I) of 0.1 microM that accounts for a maximum of 70% of the inhibition. There is also a low affinity site with a K(I) of 6 microM that accounts for a maximum of 30% inhibition. Kinetic analysis revealed that the high-affinity inhibition of the mitochondrial and microsomal LACS forms is the result of triacsin C binding at the palmitate substrate site. The high-affinity triacsin C inhibition of both the mitochondrial and microsomal LACS forms was found to require a high concentration of free Mg(2+), with the EC(50) for inhibition being 3 mM free Mg(2+). The low affinity triacsin C inhibition was also enhanced by Mg(2+). The data suggests that Mg(2+) promotes triacsin C inhibition of LACS by enhancing binding at the palmitate binding site. In contrast, the partial inhibition of the XM-ligases by triacsin C, which showed only a low-affinity component, did not require Mg(2+). PMID- 15122653 TI - Effect of alpha-tocopherol and N-acetylcysteine on benzoyl peroxide toxicity in human keratinocytes. AB - Benzoyl peroxide is a free-radical generating compound widely used in the polymer industry and also in pharmaceuticals as antimicrobial agent to treat acne. However, benzoyl peroxide causes irritation and contact dermatitis in about 1% of patients. Concern over the use of this compound is motivated by the demonstration that it can also act as skin tumor promoter in mice. In addition, benzoyl peroxide induces DNA strand breaks in many cells, including keratinocytes. Benzoyl peroxide toxicity is presumably mediated by the formation of reactive free radicals and by the consumption of intracellular antioxidants. In this work we investigated the effect of both the lipophilic antioxidant alpha-tocopherol and the hydrophilic thiol donor N-acetylcysteine (NAC) in human keratinocyte line HaCaT exposed to benzoyl peroxide. A protective effect against benzoyl peroxide cytotoxicity was achieved when cells were grown on a alpha-tocopherol layer. On the contrary, the addition of alpha-tocopherol dissolved in ethanol had a pro oxidant effect, leading to an enhancement of benzoyl peroxide toxicity. Cytotoxicity was also reduced adding NAC to the culture medium; the presence of both NAC and alpha-tocopherol exerts a synergistic cytoprotection. PMID- 15122654 TI - Adjuvant fractionated high-dose-rate intracavitary brachytherapy after external beam radiotherapy in Tl and T2 nasopharyngeal carcinoma. AB - BACKGROUND: The value of high-dose-rate intracavitary brachytherapy (HDRIB) for persistent or recurrent nasopharyngeal carcinoma has been well described; however, the benefit of routine adjuvant fractionated HDRIB following external beam radiation therapy (EBRT) has not been completely determined. The objective of this analysis was to evaluate the outcome of two fractions of adjuvant HDRIB treatment in Tl and T2 nasopharyngeal carcinoma. METHODS: Thirty-three consecutive and nonselected patients who had Tl and T2 non-disseminated nasopharyngeal carcinoma were treated according to an IRB approved institutional research protocol between March 1999 and July 2001. By the 1997 AJCC cancer staging classification, 22 patients (67%) had Tl disease and 11 patients (33%) had T2 disease. Seventeen of these patients who had stage I or stage II disease (i.e., NO or Nl) were treated with EBRT followed by two fractions of adjuvant HDRIB (group 1); 16 patients who had stage III or stage IV disease (i.e., N2 or N3) were treated with concurrent cisplatin, EBRT and adjuvant HDRIB and subsequent adjuvant cisplatin and fluorouracil (5-FU) chemotherapy (group 2). EBRT was delivered by daily conventional fractionation to a total dose of 66 Gy to the primary tumor. Nodal disease received 66 Gy if it was less than 3 cm in maximum diameter and 70 Gy if larger or there was palpable residual disease after 66 Gy. A total of 10 Gy of HDRIB in 2 equal fractions of 5 Gy spaced 1 week apart was delivered starting 1 week after the completion of EBRT. All patients were assessed for treatment response, local control, survival, and toxicity. RESULTS: The median follow up for all 29 surviving patients is 29 months (range: 17-38 months). One patient died 7 months and one died 18 months after radiation therapy from the effects of distant metastases; two died of unrelated causes. At the time of this analysis, one patient (3%) had persistent local disease and one patient (3%) developed pathologically confirmed local recurrence in the nasopharynx. In addition, one patient (3%) developed recurrence only in a neck node followed by distant metastasis, and two patients (6%) developed distant metastasis without locoregional relapse. The 2-year local control rate at the primary site was 93.6%, and the overall survival and disease-free survival rates were 82% and 74% respectively. All patients experienced some degree of acute and/or late toxicity related to radiation therapy. Ten patients (30%) experienced grade 3 acute and/or late toxicity and six patients (18%) developed grade 4 acute and/or late toxicity. No grade 5 toxicity occurred. No unexpected damage of structures within the HDRIB fields was detected. CONCLUSIONS: EBRT supplemented by two fractions of adjuvant HDRIB produced a 93.6% local control rate for Tl and T2 nasopharyngeal cancer at 2 years of follow up, with acceptable rates of acute and late toxicity. Brief adjuvant HDRIB appears to permit dose escalation safely, even in patients who receive chemotherapy concurrently with conventional radiation therapy. This strategy needs to be optimized and then tested in a prospective randomized phase III trial to learn if it can improve outcome. PMID- 15122655 TI - Angiogenesis and skin carcinomas with skull base invasion: a case-control study. AB - BACKGROUND: Some skin carcinomas may be very aggressive. Intensity of angiogenesis, measured by intratumoral vessel density using expression of CD34, has been associated with tumor aggressiveness. In this study, the expression of CD34 in basal cell carcinomas ( BCCs) and squamous cell carcinomas (SCCs) with skull base invasion was compared with that in tumors with good outcome. METHODS: Expression of CD34 was graded as mild, moderate, and intense, in 24 BCCs and 11 SCCs with skull base invasion. The control group included 23 BCCs and 10 SCCs. RESULTS: Intense expression of CD34 was noted in 25.00% of BCCs with skull base invasion, compared with 4.35% in the control group (p =.058). Regarding SCCs, intense expression of CD34 was found in 54.55% of aggressive tumors, compared with 10.00% in the control group (p =.133). CONCLUSIONS: A trend toward denser microvascular angiogenesis was observed in both BCCs and SCCs with skull base invasion compared with less aggressive controls. PMID- 15122656 TI - Numeracy and the shortcomings of utility assessment in head and neck cancer patients. AB - BACKGROUND: Because survival differences between surgical and nonsurgical treatment for head and neck cancer (HNC) are hard to detect, increasing focus has been placed on quality of life (QOL) differences after treatment. Utility assessment provides insight into QOL. Evidence suggests that a patient's comfort with numerical concepts ("numeracy") may influence utility measures. We hypothesize that patients who are nonnumerate provide inconsistent utility data in QOL studies. METHODS: New HNC (n = 18) patients were recruited to participate. Patients completed a numeracy questionnaire, a utility assessment, and a global QOL questionnaire. Higher scores reflect better function. Interviewers rated the functional level of each patient. For both numerate and nonnumerate patients, utility scores were compared with global QOL (good vs poor) and observer-rated function. RESULTS: Half of the patients were numerate. Numerate patients who rated their QOL as good had significantly higher utility scores than did patients with poor global QOL (0.95 vs 0.43, p =.03). In contrast, nonnumerate patients with good QOL had lower utility scores than did patients with poor QOL (0.45 vs 0.77, NS). Utility scores for numerate patients correlated well with observer rated function (r = 0.41 to r = 0.57), whereas those of nonnumerate patients did not (r = -0.16 to r = 0.06). CONCLUSIONS: QOL evaluation through utility assessment may provide inaccurate and contradictory data about patient functioning for nonnumerate patients. This may confound QOL assessment when interpreting utility data. PMID- 15122657 TI - Overexpression of p27BBP in head and neck carcinomas and their lymph node metastases. AB - BACKGROUND: p27(BBP) is a regulator of ribosome assembly and an essential nuclear and cytoplasmic component of eukaryotes. METHODS: We investigated the immunochemical distribution of p27(BBP) in head and neck carcinomas, in the associated normal mucosa, and in regional lymph nodes. RESULTS: p27(BBP) is detectable in mucosal cells but is overexpressed in carcinomas, highly concentrated in large polymorphous nucleoli, and even larger and more evident in lymph node metastatic foci. Western blotting confirms increased p27(BBP) in carcinomas versus normal mucosa and also in metastatic versus normal lymph nodes. The overexpression of p27(BBP) corresponds to mRNA upregulation in carcinomas. Unexpectedly, a 52-kDa band specifically reacting with antibodies to p27(BBP) was observed in several carcinomas. CONCLUSIONS: p27(BBP) alterations are common events in the transition to malignancy and are probably involved in squamous carcinoma progression. Immune reagents raised to p27(BBP) may provide additional diagnostic tools for surgical pathology of tumor boundaries and lymph nodes. The 52-kDa band may represent an abnormal form of p27(BBP) expressed by transformed airway epithelia. PMID- 15122658 TI - Accuracy of fine-needle aspiration cytology of salivary gland lesions in the netherlands cancer institute. AB - BACKGROUND: To evaluate the accuracy of fine-needle aspiration cytology (FNAC) in salivary gland lesions in a tertiary referral center. METHODS: A cytohistologic correlation study was performed using an automated pathology database of 1023 patients diagnosed with a salivary gland lesion. RESULTS: In 388 cases, both cytology and histology were available. Using cytologic confirmation of malignancy as the starting point, the sensitivity, specificity, and accuracy of FNAC in this study were 88%, 99%, and 96%, respectively. Exact type-specific concordance of the malignant diagnosis was achieved in 66 (88%) of 75 cases and in 211 (95%) of 223 benign cases. Of the 19 cases with a cytologic diagnosis "cyst," four proved to be malignant. A non-neoplastic lesion at cytology proved to be correctly classified in 53 (68%) of 80 patients. CONCLUSIONS: Our data show that cytology is a reliable and accurate technique to assess lesions of the salivary glands. The cytologic diagnosis of "cysts" and "non-neoplastic lesions" should be interpreted with caution. PMID- 15122659 TI - High-resolution imaging of the thyroid gland using optical coherence tomography. AB - BACKGROUND: Current diagnostic imaging modalities of the thyroid gland cannot reliably distinguish benign from malignant lesions, primarily because of their inability to visualize microscopic structure. A high-resolution imaging technique capable of examining thyroid tissue architectural morphology in real time is needed. Optical coherence tomography (OCT) has been shown to achieve high resolutions approaching the cellular range (1-15 microm). The feasibility of optical coherence tomography for imaging thyroid tissue was explored ex vivo on the human thyroid gland. METHODS: High-resolution OCT was performed in real time at 2 to 4 frames per second on three postmortem and 15 surgically excised thyroid glands containing normal, hyperplastic, and neoplastic tissue. OCT images acquired were compared with those obtained using standard histopathologic methods. RESULTS: The microstructure of the normal thyroid gland, including colloid-filled follicles as small as 15 microm and their supporting stroma, was clearly identified. OCT images of degenerative, hyperplastic, adenomatous, and malignant change within the thyroid gland were shown to correlate well with corresponding histopathologic findings. CONCLUSIONS: The ability of OCT to image thyroid tissue microarchitecture makes it a potentially powerful technology that can be used to assess the thyroid gland at a resolution greater than currently available clinical imaging modalities. PMID- 15122660 TI - Promising results with chemoradiation in patients with sinonasal undifferentiated carcinoma. AB - BACKGROUND: Sinonasal undifferentiated carcinoma (SNUC) is an uncommon malignancy associated with poor prognosis. The optimal treatment approach for SNUC has not been established was performed. METHODS: A retrospective review of all patients with SNUC seen at the Peter MacCallum Cancer Centre over a 12-year period. RESULTS: Ten patients with SNUC were identified, with nine having locally advanced disease (T4). Seven were treated with three cycles of platinum and 5 fluorouracil followed by radiation with two cycles of concurrent platinum. In these seven patients, the 2-year progress on-free survival was 43% (95% CI, 11% to 82%) and 2-year overall survival was 64% (95% CI, 23% to 91%). One patient with a T1N0 nasal cavity tumor treated with radiation alone has not relapsed. Two patients who were treated with initial surgical resection, prior to referral to our institution, received postoperative radiation, but they subsequently had relapses and died. CONCLUSION: Induction chemotherapy followed by concurrent chemoradiation is promising treatment strategy for SNUC. PMID- 15122661 TI - CO2 laser treatment of supraglottic cancer. AB - BACKGROUND: The aim of the study was to evaluate the effectiveness of endoscopic CO(2) laser microsurgery in the management of supraglottic cancer. METHODS: One hundred twenty-four patients (116 men and eight women; mean age, 59 years; range 31-81 years) with supraglottic cancer underwent CO(2) laser surgery (mean follow up 5 years). The study patients were classified as follows: 45 patients, T1N0M0; 61 patients, T2N0M0; and 18 patients, T3N0M0. According to the staging, the following procedures were adopted: epiglottectomy, resection of aryepiglottic fold or false vocal cord in T1 patients (group A); resection of the false vocal cord and adjacent structures in T2 patients (group B); and supraglottic laryngectomy in T3 patients (group C). Statistical comparison of survival parameters was carried out with Wilcoxon test, considering p <.05 the minimum significance value. RESULTS: Overall actuarial survival, adjusted actuarial survival, and no evidence of disease at 5 years were 91%, 97%, and 82% in group A; 88%, 94%, and 59% in group B; and 81%, 81%, and 51% in group C, respectively. The statistical analysis of survival parameters showed a significant difference in the comparison of T1 versus T2 and T2 versus T3 tumors (p <.01). Actuarial local control, actuarial nodal control, and actuarial distant metastasis control at 5 years were 82%, 82%, and 100% in T1 patients; 63%, 90%, and 98% in T2 patients; and 77%, 75%, and 93% in T3 patients. Laryngeal preservation rate was 88.6% in T1 patients, 85.4% in T2 patients, and 93.7% in T3 patients. Patients in groups A and B were discharged after 3 to 12 days, and patients in group C, were discharged after 14 to 20 days. CONCLUSIONS: The results of this study show that endoscopic CO(2) laser surgery is highly effective in the treatment of T1 and T2 supraglottic cancer. In T3 cancer, the CO(2) laser should be implemented in those cases where radical excision by endoscopic route is feasible. PMID- 15122662 TI - Neck dissection in the combined-modality therapy of patients with locoregionally advanced head and neck cancer. AB - PURPOSE: The purpose of this study was to evaluate the role of neck lymph node (ND) in the combined dissection modality therapy for locoregionally advanced head and neck. METHODS: We identified patients with N2-N3 head and neck cancers who were enrolled in three consecutive multicenter phase II studies of concurrent chemoradiotherapy utilizing 5-fluorouracil and hydroxyurea on an alternate-week schedule with radiotherapy twice daily plus either cisplatin (C-FHX) or paclitaxel (T-FHX). Patients with unknown primary tumors, nasopharyngeal or paranasal sinus primaries, nonsquamous histology, progression or death during therapy, or incomplete therapy were excluded. RESULTS: A total of 131 patients were analyzed. Seventy-nine percent had N2 stage. ND was performed in 92 patients (70%), either prior to enrollment (n = 31) or after chemoradiotherapy (n = 61). With a median follow-up of 4.6 years, the 5-year locoregional and neck progression-free survival (PFS) rates were higher in patients with ND versus patients without ND: 88% versus 74% (p =.02) and 99% versus 82% (p =.0007). respectively; there was also a trend toward improved overall survival (OS) with ND, but PFS and distant PFS were comparable. In the subset of patients with N3 disease, ND was associated not only with better locoregional control but also with improved distant PFS. However, in patients with clinical complete response (n = 92), no significant differences in PFS (68% vs 75% at 5 years, p =.53) or any other survival parameters with or without ND were observed. CONCLUSIONS: ND improves neck control and is required for patients with clinically residual disease or N3 neck cancer but has no significant impact on the outcome of patients with N2 stage disease who are rendered clinically disease-free with intensive concurrent chemoradiotherapy. PMID- 15122663 TI - Comparison of miniplates and reconstruction plates in mandibular reconstruction. AB - BACKGROUND: The aim of this study is to compare complication rates of miniplates versus reconstruction plates in the fixation of vascularized grafts into segmental mandibular defects. METHODS: Retrospective analysis of 143 consecutive successful microvascular composite flaps performed between 1993 and 2001 was performed. Data were gathered from a computerized database, case notes and pathology reports. Complications were classified as dehiscence, infection, plate or bone removal. RESULTS: In the series, 49% of patients received miniplates, and 51% received plates. No significant differences in complication rates were found between those grafts fixed with miniplates (27%) and those with reconstruction plates (30%). Plate choice was primarily determined by consultant preference. No significant differences were found in patient, defect, treatment, or follow-up characteristics between the plate groups. Twenty-nine percent of patients had at least one late complication at the reconstructed site, and this was higher (39%) in those who had postoperative radiotherapy. CONCLUSIONS: No evidence was found in this study that the increased rigidity offered by reconstruction plates influences the rate of plate or bone removal, infection, or plate exposure. Thus, the decision to use reconstruction or miniplates is not dependent on the rate of plate complications. PMID- 15122664 TI - Skull base cerebrospinal fluid fistula: a novel detection method based on two dimensional electrophoresis. AB - BACKGROUND: The detection of the presence of cerebrospinal fluid (CSF) in nasal secretions contaminated with blood and mucus remains a challenging clinical problem. METHODS: A prospective study was conducted from November 1998 to February 2002, including 42 patients. Samples (250 microL) of nasal secretions were analyzed by two-dimensional electrophoresis (2-DE). Plasma, mucus, and CSF were identified by specific proteins markers corresponding to characteristic trains of spots. Intrathecal injection of fluorescein followed by the detection of fluorescein on endoscopic examination of the nasal cavities was considered a positive reference for CSF rhinorrhea. RESULTS: In all cases of positive fluorescein test, we unambiguously observed the presence of several specific CSF markers onto the 2-DE gels. Conversely, all negative fluorescein tests were associated with the absence of CSF-specific spots. CONCLUSIONS: Two-DE analyses of biologic fluids of nasal origin should be considered as a reliable diagnostic tool in case of suspicion of CSF leak. PMID- 15122665 TI - Myoepithelioma of the soft tissue of the head and neck: a case report and review of the literature. AB - BACKGROUND: Extraglandular myoepitheliomas are neoplasms that seldom occur in the soft tissue of the head and neck region. Misdiagnosis of these neoplasms as more aggressive tumors can lead to unnecessary treatment. METHODS: We describe a myoepithelioma of cervical soft tissue. The histopathology of the tumor, its immunophenotype, its differential diagnosis, and a review of the literature are presented. RESULTS: Histopathologically, the tumor was composed of epithelioid cells with eosinophilic cytoplasm and eccentric nuclei arranged in cords and files. On immunohistochemical analysis, the cells expressed cytokeratin 14, calponin, glial fibrillary acid protein, and p63 and showed focal positivity for S-100 protein. Together, these markers identified the cells as myoepithelial type. A literature review identified only five cases of myoepithelioma in the soft tissue of the head and neck region in which detailed clinical information was provided. CONCLUSIONS: Myoepitheliomas can have cells with variable morphology arranged in different histologic patterns. Immunohistochemical analysis is crucial for unequivocal diagnosis when myoepitheliomas occur in extraglandular locations. PMID- 15122667 TI - Quantitative T1rho NMR spectroscopy of rat cerebral metabolites in vivo: effects of global ischemia. AB - The NMR relaxation times (T(1rho), T(2), and T(1)) of water, N-acetylaspartate (NAA), creatine (Cr), choline-containing compounds (Cho), and lactate (Lac) were quantified in rat brain at 4.7 T. In control animals, the cerebral T(1rho) figures, as determined with a spin-lock field of 1.0 G, were 575 +/- 30 ms, 380 +/- 19 ms, 705 +/- 53 ms, and 90 +/- 1 ms for NAA, Cr, Cho, and water, respectively. The T(1rho) figures were 62-103% longer than their respective T(2) values determined by a multiecho method. In global (ischemic) ischemia, T(1rho) of NAA declined by 34%, that of Cr and Cho did not change, and that of water increased by 10%. The T(1rho) of lactate in ischemic brain was 367 +/- 44 ms. Similar patterns of changes were observed in the multiecho T(2) of these cerebral metabolites. The T(1) of water and NAA changed in a fashion similar to that of T(1rho) and T(2). These results show differential responses in metabolite and water T(1rho) relaxation times following ischemia, and indicate that metabolite T(1rho) and T(2) relaxation times behave similarly in the ischemic brain. The contributions of dipolar and nondipolar effects on T(1rho) relaxation in vivo are discussed in this work. PMID- 15122668 TI - Robust automated shimming technique using arbitrary mapping acquisition parameters (RASTAMAP). AB - Quantitative MRI techniques as well as methods such as blood oxygen level dependent (BOLD) imaging and in vivo spectroscopy require stringent optimization of magnetic field homogeneity, particularly when using high main magnetic fields. Automated shimming approaches require a method of measuring the main magnetic field, B(0), followed by adjusting the currents in resistive shim coils to maximize homogeneity. A robust automated shimming technique using arbitrary mapping acquisition parameters (RASTAMAP) using a 3D multiecho gradient echo sequence that measures B(0) with high precision was developed. Inherent compensation and postprocessing methods enable removal of artifacts due to hardware timing errors, gradient propagation delays, gradient amplifier asymmetry, and eddy currents. This allows field maps to be generated for any field of view, bandwidth, resolution, or acquisition orientation without custom tuning of sequence parameters. Field maps of an aqueous phantom show +/- 1 Hz variation with altered acquisition orientations and bandwidths. Subsequent fitting of measured shim coil field maps allows calculation of shim currents to produce optimum field homogeneity. PMID- 15122669 TI - Human imaging of phosphorus in cortical and trabecular bone in vivo. AB - Phosphorus was imaged in vivo in human cortical and trabecular bone and the T(1) and T(2) (*) were measured. An ultrashort TE (UTE) pulse sequence (TE = 70 microm) was used with half pulse excitation and radial mapping of k-space from the center out. T(2) (*) was measured using multiple echo times and T(1) was measured both by saturation recovery and by a method using different RF pulse amplitudes. Seven normal subjects (32-85 years) were examined. Phosphorus was imaged, with a true in-plane resolution of 2.9 x 2.9 mm and a signal-to-noise ratio (SNR) of 19:1, in both cortical and trabecular bone. The mean T(2) (*) value was 207 +/- 12 micros, and the mean T(1) value was 8.6 +/- 3.0 sec. Images and measurements were obtained in realistic times on a clinical MR system. This may provide a new approach to characterizing disease of bone. PMID- 15122670 TI - Perfusion MRI of U87 brain tumors in a mouse model. AB - Continuous arterial spin labeling (CASL) was used to obtain an index of cerebral blood flow (ICBF) in the normal mouse brain and in an orthotopic mouse model of human U87 high-grade glioma at 8.5 T. Under the assumption of a constant tissue:blood partition coefficient for water in different tissues, the mean ICBF (n = 14) was found to be 50 +/- 9 mL/100g/min for tumor core and 209 +/- 11 mL/100g/min for normal tissue. The apparent T(1) (T(1app)) was 2.01 +/- 0.06 sec for tumor core and 1.66 +/- 0.03 sec for normal tissue. The ICBF and the T(1app) values were significantly different (P < 0.001) between these two regions. The detailed changes of ICBF and T(1app) in the transition from the tumor core through the tumor periphery to surrounding tissue were studied. Immunohistochemistry indicated that tumor vascularity was not uniform, with microvessel density highest in normal brain and the tissue surrounding the tumor and lowest in the tumor core. The large difference in ICBF between the tumor core and normal tissue suggests that this index might be useful for the assessment of the efficacy of antiangiogenic therapy. PMID- 15122671 TI - Determination of intracellular pH and compartmentation using diffusion-weighted NMR spectroscopy with pH-sensitive indicators. AB - The intracellular pH (pHi) of a series of cancer cell lines was determined using the pH-sensitive indicators imidazole (Im) or histidine (His) and diffusion weighted (DW) proton NMR spectroscopy. The DW method allows the observation at high magnetic field gradient values of only the slow-moving (intracellular) components, thus ensuring complete separation between intra- and extracellular components. Using the chemical shift difference (deltadelta) between the imidazole ring C2-H and C4(5)-H peaks, we were able to measure the pHi independently of chemical shift standardization. With His, the cell lines gave pHi values of approximately 6.5-7.0, whereas with Im, a second, more acidic compartment (pHi = 5.5-5.8) was also observed. An inverse correlation was also found between pHi and the intracellular lactate concentration. This method may be applicable to in vivo pH determinations. PMID- 15122672 TI - Quantitation of simulated short echo time 1H human brain spectra by LCModel and AMARES. AB - LCModel and AMARES, two widely used quantitation tools for magnetic resonance spectroscopy (MRS) data, were employed to analyze simulated spectra similar to those typically obtained at short echo times (TEs) in the human brain at 1.5 T. The study focused mainly on the influence of signal-to-noise ratios (SNRs) and different linewidths on the accuracy and precision of the quantification results, and their effectiveness in accounting for the broad signal contribution of macromolecules and lipids (often called the baseline in in vivo MRS). When applied in their standard configuration (i.e., fitting a spline as a baseline for LCModel, and weighting the first data points for AMARES), both methods performed comparably but with their own characteristics. LCModel and AMARES quantitation benefited considerably from the incorporation of baseline information into the prior knowledge. However, the more accurate quantitation of the sum of glutamate and glutamine (Glx) favored the use of LCModel. Metabolite-to-creatine ratios estimated by LCModel with extended prior knowledge are more accurate than absolute concentrations, and are nearly independent of SNR and line broadening. PMID- 15122673 TI - MRI investigation of the threshold for thermally induced blood-brain barrier disruption and brain tissue damage in the rabbit brain. AB - The ability of MRI-derived thermometry to predict thermally induced tissue changes in the brain was tested, and the thermal thresholds for blood-brain barrier (BBB) disruption and brain tissue damage were estimated. In addition, the ability of standard MRI to detect threshold-level effects was confirmed. These safety thresholds are being investigated to provide guidelines for clinical thermal ablation studies in the brain. MRI-monitored focused ultrasound heating was delivered to 63 locations in 26 rabbits. Tissue changes were detected in T(2) weighted imaging and T(1)-weighted imaging (with and without contrast) and with light microscopy. The probability for tissue damage as a function of the accumulated thermal dose, the peak temperature achieved, the applied acoustic energy, and the peak acoustic power was estimated with probit regression. The discriminative abilities of these parameters were compared using the areas under the receiver operator characteristic (ROC) curves. In MRI, BBB disruption was observed in contrast-enhanced T(1)-weighted imaging shortly after the ultrasound exposures, sometimes accompanied by changes in T(2)-weighted imaging. Two days later, changes in T(2)-weighted imaging were observed, sometimes accompanied by changes in T(1)-weighted imaging. In histology, tissue damage was seen at every location where MRI changes were observed, ranging from small (diameter <1.0 mm) areas of tissue necrosis to severe vascular damage and associated hemorrhagic infarct. In one location, small (diameter: 0.8 mm) damage was not detected in MRI. The thermal dose and peak temperature thresholds were between 12.3-40.1 equivalent min at 43 degrees C and 48.0-50.8 degrees C, respectively, and values of 17.5 equivalent min at 43 degrees C and 48.4 degrees C were estimated to result in tissue damage with 50% probability. Thermal dose and peak temperature were significantly better predictors than the applied acoustic energy and peak acoustic power (P < 0.01). BBB disruption was always accompanied by tissue damage. The temperature information was better than the applied acoustic power or energy for predicting the damage than the ultrasound parameters. MRI was sensitive in detecting threshold-level damage. PMID- 15122674 TI - Characterizing non-Gaussian diffusion by using generalized diffusion tensors. AB - Diffusion tensor imaging (DTI) is known to have a limited capability of resolving multiple fiber orientations within one voxel. This is mainly because the probability density function (PDF) for random spin displacement is non-Gaussian in the confining environment of biological tissues and, thus, the modeling of self-diffusion by a second-order tensor breaks down. The statistical property of a non-Gaussian diffusion process is characterized via the higher-order tensor (HOT) coefficients by reconstructing the PDF of the random spin displacement. Those HOT coefficients can be determined by combining a series of complex diffusion-weighted measurements. The signal equation for an MR diffusion experiment was investigated theoretically by generalizing Fick's law to a higher order partial differential equation (PDE) obtained via Kramers-Moyal expansion. A relationship has been derived between the HOT coefficients of the PDE and the higher-order cumulants of the random spin displacement. Monte-Carlo simulations of diffusion in a restricted environment with different geometrical shapes were performed, and the strengths and weaknesses of both HOT and established diffusion analysis techniques were investigated. The generalized diffusion tensor formalism is capable of accurately resolving the underlying spin displacement for complex geometrical structures, of which neither conventional DTI nor diffusion-weighted imaging at high angular resolution (HARD) is capable. The HOT method helps illuminate some of the restrictions that are characteristic of these other methods. Furthermore, a direct relationship between HOT and q-space is also established. PMID- 15122675 TI - Improved route for the visualization of stem cells labeled with a Gd-/Eu-chelate as dual (MRI and fluorescence) agent. AB - A simple labeling procedure of stem/progenitor cells based on the use of Gd HPDO3A and Eu-HPDO3A, respectively, is described. The Gd-chelate acts as T(1) agent for MRI visualization, whereas the corresponding Eu-chelate acts as reporter in fluorescence microscopy. Owing to their substantial chemical equivalence, the two chelates are equally internalized in EPCs (endothelial progenitor cells), thus allowing their visualization by both techniques. The lanthanide chelates are entrapped in endosomic vesicles and the labeled cells retain biological activity with preservation of viability and pro-angiogenesis capacity. Hyperintense spots in MR have been observed for Gd-labeled EPCs injected under mice kidney capsule or grafted on a subcutaneous Matrigel plug up to 14 days after transplantation. PMID- 15122676 TI - Quantitative description of proton exchange processes between water and endogenous and exogenous agents for WEX, CEST, and APT experiments. AB - The proton exchange processes between water and solutes containing exchangeable protons have recently become of interest for monitoring pH effects, detecting cellular mobile proteins and peptides, and enhancing the detection sensitivity of various low-concentration endogenous and exogenous species. In this work, the analytic expressions for water exchange (WEX) filter spectroscopy, chemical exchange-dependent saturation transfer (CEST), and amide proton transfer (APT) experiments are derived by the use of Bloch equations with exchange terms. The effects of the initial states for the system, the difference between a steady state and a saturation state, and the relative contributions of the forward and backward exchange processes are discussed. The theory, in combination with numerical calculations, provides a useful tool for designing experimental schemes and assessing magnetization transfer (MT) processes between water protons and solvent-exchangeable protons. As an example, the case of endogenous amide proton exchange in the rat brain at 4.7 T is analyzed in detail. PMID- 15122677 TI - Comparison of BOLD contrast and Gd-DTPA dynamic contrast-enhanced imaging in rat prostate tumor. AB - The microcirculation and oxygenation of a tumor play important roles in its responsiveness to cytotoxic treatment, and noninvasive assessments of its vascular properties may have prognostic value. Dynamic contrast-enhanced (DCE) (1)H MRI based on infusion of Gd-DTPA, and blood oxygen level-dependent (BOLD) contrast based on altering inhaled gas are both sensitive to vascular characteristics. This study compares the effects observed in eight Dunning prostate R3327-AT1 rat tumors imaged sequentially at 4.7 Tesla by echo-planar imaging (EPI). Both interventions generated a significant response, and each revealed significant differences between the center and periphery of the tumors. On a voxel-by-voxel basis across the whole tumor population, there was a close correlation between the maximum rate of signal response and the magnitude of response to each intervention (R(2) >or= 0.6, P < 0.0001). However, when the data were analyzed separately for each individual tumor, some showed a weak correlation (R(2) < 0.4), particularly for DCE, and the nature (slope) varied between separate tumors. Generally, there was a weak correlation (N = 7, R(2) < 0.5) between responses to the two interventions on a tumor-by-tumor basis, which emphasizes that the techniques are not equivalent. Both techniques revealed intra and intertumor heterogeneity, but the BOLD response was more rapidly reversible than the DCE response. This suggests that the BOLD technique may be a useful tool for investigating interventions (such as drugs) that cause vascular disruption. PMID- 15122678 TI - Measuring blood volume and vascular transfer constant from dynamic, T(2)* weighted contrast-enhanced MRI. AB - Dynamic, contrast-enhanced MRI (deMRI) is increasingly being used to evaluate cerebral microcirculation. There are two different approaches for analyzing deMRI data. Intravascular indicator dilution theory has been used to estimate blood volume (and perfusion), usually from T(2)- or T(2) (*)-weighted images of the first pass of the bolus. However, the theory assumes that the tracer (i.e., contrast agent) remains intravascular, which is often not the case when the blood brain barrier (BBB) is damaged. Furthermore, the method provides no information on the vascular transfer constant. Pharmacokinetic modeling analyses of T(1) weighted images after first pass do give values of the vascular transfer constant and the volume of the extravascular, extracellular space (EES), but they generally are unable to give estimates of blood volume. In this study we apply pharmacokinetic modeling to dynamic T(2) (*)-weighted imaging of the first pass of a tracer bolus. This method, which we call first-pass pharmacokinetic modeling (FPPM), gives an estimate of the blood volume, vascular transfer constant, and EES volume. The method was applied to a group of 26 patients with surgically proven tumors (10 glioblastomas multiforme (GBMs), six lymphomas, and 10 meningiomas). The measurements of the blood volume and transfer constant were consistent with the known physiology of these tumors. PMID- 15122679 TI - Estimation of the differential pressure at renal artery stenoses. AB - Atherosclerotic disease of the renal artery can lead to reduction in arterial caliber and ultimately to conditions including renovascular hypertension. Renal artery stenosis is conventionally assessed, using angiography, according to the severity of the stenosis. However, the severity of a stenosis is not a reliable indicator of functional significance, or associated differential pressure, of a stenosis. A methodology is proposed for estimation of the renal artery differential pressure (RADP) from MR imaging. Realistic computational fluid dynamics (CFD) models are constructed from MR angiography (MRA) and phase contrast (PC) MR. The CFD model is constructed in a semiautomated manner from the MR images using the Isosurface Deformable Model (IDM) for surface reconstruction and a Marching Front algorithm for construction of the volumetric CFD mesh. Validation of RADP estimation was performed in a realistic physical flow-through model. Under steady flow, the CFD estimate of the differential pressure across a stenosis in the physical flow-through model differed by an average of 5.5 mmHg from transducer measurements of the pressure differential, for differential pressures less than 60 mmHg. These results demonstrate that accurate estimates of differential pressure at stenoses may be possible based only on structural and flow images. PMID- 15122680 TI - Direct CSF injection of MnCl(2) for dynamic manganese-enhanced MRI. AB - MnCl(2) was injected intrathecally through the cisterna magna in rats, allowing infusion of divalent manganese ions (Mn(++)) into the CSF space and thence into the brain, without breaking the blood-brain barrier (BBB). Mn(++) uptake and washout dynamics in the brain were measured by serial T(1)-weighted MRI and EPI T(1) and T(2) mapping for up to 3 weeks after injection. Observations within the first 6 hr after injection demonstrated anterograde and bilateral distribution of the Mn(++) within the CSF space, from the olfactory bulb and frontal cortex to the brain stem. Enhancement increased in most brain areas up to 4 days after injection, and then slowly decreased. Relaxation maps at each time point demonstrated higher concentrations of Mn in basal ganglia. Residual concentrations were still observable after 3 weeks in all brain regions. With the use of MnCl(2) calibration phantoms, the maximum Mn concentration in the brain was estimated to be approximately 27 +/- 16 microM, corresponding to changes in relaxation rates of 0.49 +/- 0.30 s(-1) for R(1) and 3.9 +/- 2.4 s(-1) for R(2). For comparison, an intrathecal GdDTPA injection was performed. This injection showed different distribution dynamics: it remained chiefly within the CSF spaces, and was largely washed out after 1 day. This method shows promise as a means of supplying Mn(++) uniformly to the whole brain for a variety of chronic functional activation studies. PMID- 15122681 TI - Visualization and tracking of an inflatable balloon catheter using SSFP in a flow phantom and in the heart and great vessels of patients. AB - Passive catheter tracking involves direct interaction between the device and its surroundings, creating a local signal loss or enhancement of the image. Using only standard balloon catheters filled with CO(2) and imaged with a steady-state free precession sequence, it was possible to visualize and passively track catheters in a flow phantom and in the heart and great vessels of 20 patients without any additional image processing. The phantom work demonstrated that it was advantageous to sacrifice spatial resolution in order to increase temporal resolution. Frame rates greater than 10/sec were necessary for ease of catheter manipulation. Although only the tip of the catheter was visualized, this technique proved to be effective in patients undergoing cardiac catheterization. PMID- 15122682 TI - Selection of the optimum b factor for diffusion-weighted magnetic resonance imaging assessment of ischemic stroke. AB - The purpose of this study was to determine the diffusion sensitivity factor b that optimizes the contrast-to-noise ratio (CNR) for both diffusion-weighted signal intensity and the apparent diffusion coefficient (ADCNR) when evaluating ischemic stroke by diffusion-weighted MRI. The relative contrast, noise levels, CNR, and ADCNR were calculated for typical ADC values in human brain, 780 microm(2)/s in adults and 1200 microm(2)/s in neonates in normal tissue, 20-40% less in acute and subacute stroke, and 50% more in chronic stroke. The optimum b factor depends strongly on the ADC, whether TE is fixed or varies with the b factor, whether CNR or ADCNR is measured, and anisotropy. The optimum b factor in adults is 1000 s/mm(2) in acute and chronic stroke, and 1200 s/mm(2) in subacute stroke. The optimum values are about 200 s/mm(2) lower in neonates than in adults. The CNR and ADCNR are within 10% of the optimum over at least a 2-fold range of b factors, from 68-136% of the optimum b factor. If a single b factor is to be used for all situations, a diffusion b factor of 1000 s/mm(2) is recommended. PMID- 15122683 TI - Magnetization tagging decay to measure long-range (3)He diffusion in healthy and emphysematous canine lungs. AB - Spatial modulation (tagging) of the longitudinal magnetization allows diffusive displacements to be measured over times approximately as long as T(1) and over correspondingly long distances. Magnetization tagging is used here with hyperpolarized (3)He gas in canine lungs with unilateral elastase-induced emphysema. A new scheme for analyzing images subsequent to tagging determines the spatially-resolved fractional modulation and its decay rate, using a sliding window. The diffusivity so determined over seconds and centimeter lengths, D(sec), is smaller in all cases than the diffusivity measured over milliseconds and hundreds of microns, D(msec) (in healthy lungs, this ratio is about 0.1). While D(msec) is sensitive to lung microstructure on the alveolar level, D(sec) reflects airway connectivity and provides new information on lung structure. The results show substantial increases in D(sec) in the lungs of four dogs with clear evidence of emphysema. For these dogs, the fractional increase in long-range diffusivity D(sec) in the emphysematous lungs was greater than that in short range diffusivity D(msec). PMID- 15122684 TI - Assessment of human pulmonary function using oxygen-enhanced T(1) imaging in patients with cystic fibrosis. AB - Indirect qualitative MRI of pulmonary function is feasible using the paramagnetic effects of oxygen physically dissolved in blood. In this study, a more quantitative oxygen-enhanced pulmonary function test based on the slope of a plot of R(1) vs. oxygen concentration-the oxygen transfer function (OTF)-was developed and tested in a pool of five healthy volunteers and five patients with cystic fibrosis (CF). The lung T(1) relaxation rate, R(1), under normoxic conditions (room air, 21% O(2)), and the response to various hyperoxic conditions (40%-100% O(2)) were studied. Lung T(1) in healthy volunteers showed a relatively homogeneous distribution while they breathed room air, and a homogeneous decrease under hyperoxic conditions. Lung T(1) in CF patients showed an inhomogeneous distribution while they breathed room air, and the observed lung T(1) decrease under hyperoxia depended on the actual state of the diseased lung tissue. In the selected group of CF patients, areas with reduced OTF also showed reduced perfusion, as confirmed by qualitative contrast-enhanced MR pulmonary perfusion imaging. The results demonstrate that this completely noninvasive oxygen-enhanced pulmonary function test has potential for clinical applications in the serial diagnosis of lung diseases such as CF. . PMID- 15122685 TI - Calculation of the renal perfusion and glomerular filtration rate from the renal impulse response obtained with MRI. AB - The aim of this study was to assess the importance of deconvolution for the calculation of renal perfusion and glomerular filtration rate (GFR) on the basis of concentration-time curves as measured with perfusion MRI. Six rabbits were scanned dynamically after injection of a gadolinium chelate. Concentration-time curves were generated by manually drawing regions of interest in the aorta and the renal cortex. To remove the dependency on the arterial input function, a regularized structured total least-squares deconvolution algorithm was used to calculate the renal impulse response. This curve was fitted by the sum of two gamma variate functions, corresponding to the passage of the contrast agent in the glomeruli and the proximal convoluted tubules. Tracer kinetics models were applied to these two functions to obtain the renal perfusion and GFR. For comparison, these two parameters were also calculated on the basis of the renal concentration-time curve before deconvolution. The renal perfusion values correlated well (r = 0.9, P = 0.014) with the values calculated by a validated upslope method. The GFR values correlated well (r = 0.9, P = 0.014) with the values obtained from the clearance of (51)Cr-EDTA. A comparison of the values obtained with and without deconvolution demonstrated the necessity of deconvolution. PMID- 15122686 TI - In vivo precision of quantitative shoulder cartilage measurements, and changes after spinal cord injury. AB - Recent advances in MRI have enabled the quantitative assessment of articular cartilage morphology in human joints. In this study, we tested the hypothesis that the precision of quantitative shoulder cartilage measurements is sufficient to detect changes between and within patients, and that shoulder cartilage thickness in paraplegic patients increases due to increased loading. We imaged the shoulders of seven healthy volunteers four times using a coronal 3D, fat suppressed, gradient-echo sequence. The humeral head cartilage in seven paraplegic patients was evaluated soon after injury and 1 year post injury. A precision of 4.5% (root mean square (RMS) average coefficient of variation (CV) %) was found for shoulder cartilage thickness measurements in the humeral head. Whereas a significant decrease of cartilage thickness (-11%, P < 0.05) was observed in the knee, there was no significant change in articular cartilage thickness in the shoulder (-1.1%). Our data show, for the first time, that articular cartilage of the humeral head can be quantified with acceptable precision in vivo. It was demonstrated that, in contrast to the knee, the articular cartilage morphology of the humeral head changes very little (i.e., there is no significant increase or decrease in thickness) after spinal cord injury (SCI). PMID- 15122687 TI - Reduced RF power without blurring: correcting for modulation of refocusing flip angle in FSE sequences. AB - In order to reduce the RF power deposition of fast spin echo sequences operated at high field strength, the flip angles of the refocusing pulse train are varied from pulse to pulse using a modulated angle refocusing train method. The technique employs high flip angle pulses prior to sampling the center of k-space in order to preserve T(2) contrast, low flip angles after sampling the center of k-space to reduce power and prolong relaxation, and a smooth transition between the high and low flip angle regimes in order to maintain the pseudosteady-state, maximizing signal and avoiding artifact-inducing oscillations. An analytical expression is used to predict and correct for the flip angle dependence of the signal, thus eliminating any deleterious effects of flip angle modulation on the point spread function. Analysis of resolution and SNR were performed in simulation and phantom studies. In human imaging studies, it is shown that RF energy deposition per slice in a single-shot fast spin echo application can be reduced by up to 75%, making the sequence as practical at 3 T as it is has been at 1.5 T. PMID- 15122688 TI - Analysis of multiple-acquisition SSFP. AB - Refocused steady-state free precession (SSFP) is limited by its high sensitivity to local field variation, particularly at high field strengths or the long repetition times (TRs) necessary for high resolution. Several methods have been proposed to reduce SSFP banding artifact by combining multiple phase-cycled SSFP acquisitions, each differing in how individual signal magnitudes and phases are combined. These include maximum-intensity SSFP (MI-SSFP) and complex-sum SSFP (CS SSFP). The reduction in SSFP banding is accompanied by a loss in signal-to-noise ratio (SNR) efficiency. In this work a general framework for analyzing banding artifact reduction, contrast, and SNR of any multiple-acquisition SSFP combination method is presented. A new sum-of-squares method is proposed, and a comparison is performed between each of the combination schemes. The sum-of squares SSFP technique (SOS-SSFP) delivers both robust banding artifact reduction and higher SNR efficiency than other multiple-acquisition techniques, while preserving SSFP contrast. PMID- 15122689 TI - Effects of, and corrections for, cross-term interactions in Q-space MRI. AB - The present study assesses the effects of cross-term interactions between diffusion and imaging gradients in magnetic resonance imaging q-space analysis, and corrects for those effects for both spin echo and stimulated echo diffusion weighted sequences. These corrections are demonstrated experimentally in unrestricted media for water and theoretically by simulating the case of restricted diffusion in a sphere. By correcting for the cross-term interactions, large imaging gradients can be used without compromising the results. Ignoring cross-term interactions could lead to a misunderstanding of the q-space analysis; for instance, the microstructural size of the sample could be overestimated, or isotropic media could be misinterpreted as being anisotropic. PMID- 15122690 TI - Fast, three-dimensional free-breathing MR imaging of myocardial infarction: a feasibility study. AB - Imaging delayed hyperenhancement of myocardial infarction is most commonly performed using an inversion recovery (IR) prepared 2D breathhold segmented k space gradient echo (FGRE) sequence. Since only one slice is acquired per breathhold in this technique, 12-16 successive breathholds are required for complete anatomical coverage of the heart. This prolongs the overall scan time and may be exhausting for patients. A navigator-echo gated, free-breathing, 3D FGRE sequence is proposed that can be used to acquire a single slab covering the entire heart with high spatial resolution. The use of a new variable sampling in time (VAST) acquisition scheme enables the entire 3D volume to be acquired in 1.5 2 min, minimizing artifacts from bulk motion and diaphragmatic drift and contrast variations due to contrast media washout. PMID- 15122691 TI - MRI monitoring of heating produced by ultrasound absorption in the skull: in vivo study in pigs. AB - The purpose of this study was to test the utility of MR thermometry for monitoring the temperature rise on the brain surface and in the scalp induced by skull heating during ultrasound exposures. Eleven locations in three pigs were targeted with unfocused ultrasound exposures (frequency = 690 kHz; acoustic power = 8.2-16.5 W; duration = 20 s). MR thermometry (a chemical shift technique) showed an average temperature rise in vivo of 2.8 degrees C +/- 0.6 degrees C and 4.4 degrees C +/- 1.4 degrees C on the brain surface and scalp, respectively, at an acoustic power level of 10 W. The temperature rise on the scalp agreed with that measured with a thermocouple probe inserted adjacent to the skull (average temperature rise = 4.6 degrees C +/- 1.0 degrees C). Characterization of the transducer showed that the average acoustic intensity was 1.3 W/cm(2) at an acoustic power of 10 W. The ability to monitor the temperature rise next to the skull with MRI-based thermometry, as shown here, will allow for safety monitoring during clinical trials of transcranial focused ultrasound. PMID- 15122692 TI - Improvement in breast lesion characterization with dynamic contrast-enhanced MRI using pharmacokinetic modeling and bookend T(1) measurements. AB - Dynamic contrast-enhanced breast MR imaging was performed on 14 patients (five cancerous lesions, nine benign) with slice-selective spoiled gradient-recalled echo (2D SPGR) imaging. Adiabatic saturation recovery T(1) measurements were performed before (T(1pre)) and after (T(1post)) 2D SPGR imaging. These two "bookend" T(1) measurements were used to calibrate the equations which were employed to convert the time course of the 2D SPGR signal strength to T(1)-vs. time, which in turn was used to compute the gadolinium concentration-vs.-time ([C](t)) in the lesion. The extraction-flow product (EF) was computed for each lesion by pharmacokinetic modeling of [C](t). For this study, EF provided a sensitivity and specificity for cancer of 100% and 78%, respectively. When only T(1pre) was used to estimate [C](t) (which assumes a priori knowledge of the shape and amplitude of the slice profile), the sensitivity and specificity fell to 80% and 56%, respectively. This is presumably due to unexpected variations in the shape and/or amplitude of the slice profile, which could be caused by factors such as patient-to-patient variations in breast geometry or inconsistently set transmit gains. Therefore, both T(1pre) and T(1post) measurements are necessary for optimum sensitivity and specificity using pharmacokinetic analysis. PMID- 15122693 TI - Artifact reduction in undersampled projection reconstruction MRI of the peripheral vessels using selective excitation. AB - The projection reconstruction (PR)-HyperTRICKS (time resolved imaging of contrast kinetics) acquisition integrates the benefits of through-plane Cartesian slice encoding and in-plane undersampled PR. It provides high spatial resolution both in-plane (about 1 mm(2)) and through-plane (1-2 mm), as well as relatively high temporal resolution (about 0.25 frames per second). However, undersampling artifacts that originate from anatomy superior or inferior to a coronal imaging FOV may severely degrade the image quality. In coronal MRA acquisitions, the slice coverage is limited in order to achieve high temporal resolution. In this report we describe an artifact reduction method that uses selective excitation in PR-HyperTRICKS. This technique significantly reduces undersampling streak artifacts while it increases the slice coverage. PMID- 15122694 TI - Quantifying arbitrary magnetic susceptibility distributions with MR. AB - Magnetic susceptibility, as a physical property of materials, plays important roles in many physical, chemical, engineering, and medical applications. Its quantification becomes of significant interest when MRI becomes a commonly used technique in biomedical applications. A general method is presented here for quantifying arbitrary magnetic susceptibility distributions in a localized region on the basis of first principles of magnetic induction field distributions in space. A proof of the concept was demonstrated by computer simulations. The study establishes the methodological basis for quantitative magnetic susceptibility imaging with MR. PMID- 15122695 TI - Method for rapid MRI needle tracking. AB - A new method for MRI needle tracking within a given two-dimensional (2D) image slice is presented. The method is based on k-space investigation of the difference image between the current dynamic frame and a reference frame. Using only a few central k-lines of the difference image and a nonlinear optimization procedure, one can resolve the parameters that define the 2D sinc function that best characterizes the needle in k-space. The spatial location and orientation of the needle are determined from these parameters. Rapid needle tracking is obtained by repeated acquisitions of the same set of several central k-lines (as in a "keyhole" protocol) and repeated computation of these parameters. The calculated needle tip is depicted on the reference image by means of a graphic overlay. The procedure was tested in computer simulations and in actual MRI scans (the computations were done offline). It was demonstrated that six k-lines out of 128 usually suffice to locate the needle. The refresh rate of the needle location depends on the time required to sample the subset of k-lines, calculate the current needle location, and refresh the reference image. PMID- 15122696 TI - Measuring the effects of indomethacin on changes in cerebral oxidative metabolism and cerebral blood flow during sensorimotor activation. PMID- 15122698 TI - Axonal degeneration in multiple sclerosis: is it time for neuroprotective strategies? PMID- 15122699 TI - Presenilin mutations associated with fronto-temporal dementia. PMID- 15122700 TI - Axonal protection using flecainide in experimental autoimmune encephalomyelitis. AB - Axonal degeneration is a major cause of permanent neurological deficit in multiple sclerosis (MS), but no current therapies for the disease are known to be effective at axonal protection. Here, we examine the ability of a sodium channel blocking agent, flecainide, to reduce axonal degeneration in an experimental model of MS, chronic relapsing experimental autoimmune encephalomyelitis (CR EAE). Rats with CR-EAE were treated with flecainide or vehicle from either 3 days before or 7 days after inoculation (dpi) until termination of the experiment at 28 to 30 dpi. Morphometric examination of neurofilament-labeled axons in the spinal cord of CR-EAE animals showed that both flecainide treatment regimens resulted in significantly higher numbers of axons surviving the disease (83 and 98% of normal) compared with controls (62% of normal). These findings indicate that flecainide and similar agents may provide a novel therapy aimed at axonal protection in MS and other neuroinflammatory disorders. PMID- 15122701 TI - A novel presenilin 1 mutation associated with Pick's disease but not beta-amyloid plaques. AB - Familial forms of frontotemporal dementia (FTD) with tauopathy are mostly caused by mutations in the gene encoding the microtubule-associated protein tau (MAPT). However, rare forms of familial tauopathy without MAPT mutations have been reported, suggesting other tauopathy-related genetic defects. Interestingly, two presenilin 1 (PS1) mutations (Leu113Pro and insArg352) recently have been associated with familial FTD albeit without neuropathological confirmation. We report here a novel PS1 mutation in a patient with Pick-type tauopathy in the absence of extracellular beta-amyloid deposits. The mutation is predicted to substitute Gly-->Val at codon position 183 (Gly183Val) and to affect the splice signal at the junction of the sixth exon and intron. Further clinical-genetic investigation showed a positive family history of FTD-like dementia and suggested that Gly183Val is associated with a phenotypically heterogeneous neurodegenerative disorder. Our results suggest PS1 as a candidate gene for Pick type tauopathy without MAPT mutations. PMID- 15122702 TI - Expression of CCR7 in multiple sclerosis: implications for CNS immunity. AB - It is unclear how immune cells traffic between the lymphoid compartment and the central nervous system (CNS), which lacks lymphatic vessels and is shielded by the blood-brain barrier. We studied the expression of CCR7, a chemokine receptor required for migration of T cells and dendritic cells (DCs) to lymphoid organs, in the CNS of patients with multiple sclerosis (MS) to gain insight into pathways for CNS immune cell trafficking. Inflamed MS lesions contained numerous CCR7+ myeloid cells expressing major histocompatibility complex class II, CD68 and CD86, consistent with maturing DCs. CCR7+ DCs also were identified in cerebrospinal fluid (CSF). These observations suggested that the afferent limb of CNS immunity is comprised, in part, of DCs, which are generated within the CNS and migrate to deep cervical lymph nodes through the CSF after antigen capture. Ninety percent of CSF T cells expressed CCR7 and CSF from patients with MS was relatively depleted of CCR7-negative effector-memory T cells. In contrast, all T cells in parenchymal MS lesions lacked CCR7, indicating local retention and differentiation of central-memory T cells upon restimulation by antigen within the CNS. These data suggested that the efferent limb of CNS immunity is executed by central-memory T cells, which enter CSF directly from the circulation. PMID- 15122703 TI - Unilateral nerve injury produces bilateral loss of distal innervation. AB - There are no known anatomical connections between neurons that innervate homologous right and left body parts. Nevertheless, some patients develop bilateral abnormalities after unilateral injury, a phenomenon often unrecognized and not yet characterized. Therefore, we examined in rats the effects of ligating and cutting one tibial nerve on sensory function and on density of innervation in hind paws contralaterally as well as ipsilaterally to the injury, at times between 1 day and 5 months after surgery. Punches removed from tibial- or sural innervated planter paw skin were immunolabeled to quantitate epidermal nerve endings. Naive and sham-operated rats provided controls. Axotomized rats had near total loss of PGP9.5(+) innervation within ipsilateral tibial-innervated skin at all time-points. Adjacent ipsilateral sural-innervated skin had persistent hyperalgesia without denervation, and robust axonal sprouting at 5 months after surgery. Contralesional hind paws lost 54% of innervation in tibial-innervated epidermis starting 1 week after surgery and persisting throughout. Contralesional sural-innervated skin had neither neurite loss nor sprouting. These results imply that unilateral nerve injury can cause profound, long lasting, nerve-branch specific loss of distal innervation contralaterally as well as ipsilaterally. They discredit the practice of using tissues contralateral to an injury to provide normative controls and suggest the possibility of rapid, transmedian postinjury signals between homologous mirror-image neurons. PMID- 15122704 TI - Cu2+ toxicity inhibition of mitochondrial dehydrogenases in vitro and in vivo. AB - Wilson's disease results from mutations in the P-type Cu(2+)-ATPase causing Cu(2+) toxicity. We previously demonstrated that exposure of mixed neuronal/glial cultures to 20 microM Cu(2+) induced ATP loss and death that were attenuated by mitochondrial substrates, activators, and cofactors. Here, we show differential cellular sensitivity to Cu(2+) that was equalized to 5 microM in the presence of the copper exchanger/ionophore, disulfiram. Because Cu(2+) facilitates formation of oxygen radicals (ROS) which inhibit pyruvate dehydrogenase (PDH) and alpha ketoglutarate dehydrogenase (KGDH), we hypothesized that their inhibition contributed to Cu(2+)-induced death. Toxic CU(2+) exposure was accompanied by early inhibition of neuronal and hepatocellular PDH and KGDH activities, followed by reduced mitochondrial transmembrane potential, DeltaPsi(M). Thiamine (1-6 mM), and dihydrolipoic acid (LA, 50 microM), required cofactors for PDH and KGDH, attenuated this enzymatic inhibition and subsequent death in all cell types. Furthermore, liver PDH and KGDH activities were reduced in the Atp7b mouse model of Wilson's disease prior to liver damage, and were partially restored by oral thiamine supplementation. These data support our hypothesis that Cu(2+)-induced ROS may inhibit PDH and KGDH resulting in neuronal and hepatocellular death. Therefore, thiamine or lipoic acid may constitute potential therapeutic agents for Wilson's disease. PMID- 15122705 TI - Magnetic resonance imaging of labeled T-cells in a mouse model of multiple sclerosis. AB - Multiple sclerosis (MS) is a T cell-mediated autoimmune disease with early lesions characterized by mononuclear cellular infiltrate, edema, demyelination, and axonal loss that contribute to the clinical course of the disease. Experimental autoimmune encephalomyelitis (EAE) in the mouse is a valuable model with a similar disease course to relapsing-remitting MS. The ability to detect the migration of encephalitogenic T cells into the central nervous system in EAE and MS would provide key information on these cells role in the development of lesions observed on magnetic resonance imaging (MRI). T cells were labeled for detection by magnetic resonance imaging using Food and Drug Administration approved, superparamagnetic iron oxide nanoparticles (Ferumoxides) complexed to poly-L-Lysine (FE-PLL). EAE was induced by adoptive transfer of either labeled or unlabeled T cells. After disease onset, FE-PLL-labeled T cells were detected in the mouse spinal cord using in vivo and ex vivo cellular MRI. Excellent correlation was seen between MRI-visible lesions in the spinal cord and histopathology. The results demonstrate that T cells labeled with FE-PLL can induce EAE disease and can be detected in vivo in the mouse model. The magnetic labeling of cells opens the possibility of monitoring specific cellular phenotypes or pharmacologically or genetically engineered cells by MRI. PMID- 15122706 TI - Vascular endothelial growth factor mediates vasogenic edema in acute lead encephalopathy. AB - Brain injury from inorganic Pb(2+) is considered the most important environmental childhood health hazard worldwide. The microvasculature of the developing brain is uniquely susceptible to high level Pb(2+) toxicity (ie, Pb(2+) encephalopathy) characterized by cerebellar hemorrhage, increased blood-brain barrier permeability, and vasogenic edema. However, the specific molecular mediators of Pb(2+) encephalopathy have been elusive. We found that Pb(2+) induces vascular endothelial growth factor/vascular permeability factor (VEGF) in cultured astrocytes (J Biol Chem, 2000;275:27874-27882). The study presented here asks if VEGF dysregulation contributes mechanistically to Pb(2+) encephalopathy. Neonatal rats exposed to 4% Pb-carbonate develop the histopathological features of Pb(2+) encephalopathy seen in children. Cerebellar VEGF expression increased approximately twofold (p < 0.01) concurrent with the development of cerebellar microvascular hemorrhage, enhanced vascular permeability to serum albumin, and vasogenic cerebellar edema (p < 0.01). No change in VEGF expression occurred in cerebral cortex that does not develop these histopathological complications of acute Pb(2+) intoxication. Pb(2+) exposure increased phosphorylation of cerebellar Flk-1 VEGF receptors and the Flk-1 inhibitor CEP-3967 completely blocked cerebellar edema formation without affecting microhemorrhage formation or blood-brain barrier permeability. This establishes that Pb(2+)-induced vasogenic edema formation develops via a Flk-1-dependent mechanism and suggests that the vascular permeability caused by Pb(2+) is Flk-1 independent. PMID- 15122707 TI - Prostanoids, not reactive oxygen species, mediate COX-2-dependent neurotoxicity. AB - The prostaglandin synthesizing enzyme cyclooxygenase-2 (COX-2) has emerged as a critical pathogenic factor in brain diseases associated with activation of N methyl-D-aspartate (NMDA) receptors, including stroke and neurodegenerative diseases. However, the COX-2 reaction products responsible for these deleterious effects have not been identified. In particular, the relative contribution to the neurotoxicity of COX-2-derived prostanoids and reactive oxygen species has not been defined. We found that the brain damage produced by direct injection of NMDA into the somatosensory cortex is attenuated by the COX-2 inhibitor NS-398 or in COX-2-null mice, but that the associated production of free radicals is not. Furthermore, COX-2 inhibition reduces the lesions even if the deleterious effects of free radicals are eliminated by the scavenger superoxide dismutase. The protection exerted by NS-398 is counteracted by a stable analog of prostaglandin E2. The findings directly implicate COX-2-derived prostanoids, rather then radicals, in the COX-2-dependent component of the damage mediated by NMDA receptors and strengthen the rationale for using COX-2 inhibitors in the treatment of neurological diseases associated with glutamate neurotoxicity. PMID- 15122708 TI - Desmin-related myopathy with Mallory body-like inclusions is caused by mutations of the selenoprotein N gene. AB - Desmin-related myopathies (DRMs) are a heterogeneous group of muscle disorders, morphologically defined by intrasarcoplasmic aggregates of desmin. Mutations in the desmin and the alpha-B crystallin genes account for approximately one third of the DRM cases. The genetic basis of the other forms remain unknown, including the early-onset, recessive form with Mallory body-like inclusions (MB-DRMs), first described in five related German patients. Recently, we identified the selenoprotein N gene (SEPN1) as responsible for SEPN-related myopathy (SEPN-RM), a unique early-onset myopathy formerly divided in two different nosological categories: rigid spine muscular dystrophy and the severe form of classical multiminicore disease. The finding of Mallory body-like inclusions in two cases of genetically documented SEPN-RM led us to suspect a relationship between MB-DRM and SEPN1. In the original MB-DRM German family, we demonstrated a linkage of the disease to the SEPN1 locus (1p36), and subsequently a homozygous SEPN1 deletion (del 92 nucleotide -19/+73) in the affected patients. A comparative reevaluation showed that MB-DRM and SEPN-RM share identical clinical features. Therefore, we propose that MB-DRM should be categorized as SEPN-RM. These findings substantiate the molecular heterogeneity of DRM, expand the morphological spectrum of SEPN-RM, and implicate a necessary reassessment of the nosological boundaries in early onset myopathies. PMID- 15122709 TI - The urokinase receptor is overexpressed in the AIDS dementia complex and other neurological manifestations. AB - The urokinase-type plasminogen activator (uPA) and its receptor (uPAR) play an important role in extracellular matrix degradation and cell migration in the central nervous system (CNS). To investigate the role of the uPA/uPAR system in the pathophysiology of acquired immunodeficiency syndrome dementia complex (ADC), we measured soluble uPAR (suPAR) levels in cerebrospinal fluid (CSF) and plasma from human immunodeficiency virus (HIV)-1-infected patients and controls. CSF suPAR levels were significantly higher in HIV-1-infected patients than in controls and in patients with ADC or opportunistic CNS infections (CNS-OIs) than in neurologically asymptomatic patients, irrespective of HIV-1 disease stage. The highest levels of suPAR were found in patients with ADC, and among those with CNS OIs in patients with cytomegalovirus encephalitis or cryptococcosis. Plasma suPAR levels were higher in HIV-1-infected patients than in controls and increased with HIV-1 disease stage regardless of the presence of CNS disease. In patients with ADC or CNS-OIs, CSF suPAR levels correlated with CSF HIV-1 RNA, but not with plasma suPAR concentrations. Highly active antiretroviral therapy was associated with a significant and parallel decrease of both CSF suPAR and HIV-1 RNA. In brain tissue from patients with HIV-1 encephalitis, uPAR was highly expressed by microglial and multinucleated giant cells staining positively for HIV-1. The overexpression of uPAR in the CNS of patients with ADC suggests that the uPA/uPAR system may contribute to the tissue injury and neuronal damage in this disease. PMID- 15122710 TI - Vigabatrin, the GABA-transaminase inhibitor, damages cone photoreceptors in rats. AB - Epileptic patients experienced an irreversible loss of their peripheral visual field upon treatment with vigabatrin (gamma-vinyl GABA), an inhibitor of the GABA degrading enzyme, GABA transaminase. Subsequently, central visual function was reported to also be irreversibly altered. This visual loss is associated with a decrease in the electroretinogram measurement localizing the deficit to the retina. To investigate its cellular origin, we treated rats daily with vigabatrin for 45 days. Two days after arresting this treatment, rats exhibited an irreversible decrease in the photopic electroretinogram, the flicker response, and the oscillatory potentials. These functional alterations were associated with a peripheral disorganization of the outer retina. However, photoreceptor damage was not limited to these disorganized areas, but cone inner and outer segments were severely injured in more central areas and their numbers were irreversibly decreased by 17 to 20%. Ultrastructural examination of the retina confirmed the presence of major photoreceptor damages, which were further supported by terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling (TUNEL) and caspase-3 activation both indicative of photoreceptor apoptosis. This study suggests that the visual field loss in vigabatrin-treated epileptic patients may result from a sequence of events starting from cone cell injury to a more severe disorganization of the photoreceptor layer. PMID- 15122711 TI - POLG mutations associated with Alpers' syndrome and mitochondrial DNA depletion. AB - Alpers' syndrome is a fatal neurogenetic disorder first described more than 70 years ago. It is an autosomal recessive, developmental mitochondrial DNA depletion disorder characterized by deficiency in mitochondrial DNA polymerase gamma (POLG) catalytic activity, refractory seizures, neurodegeneration, and liver disease. In two unrelated pedigrees of Alpers' syndrome, each affected child was found to carry a homozygous mutation in exon 17 of the POLG locus that led to a Glu873Stop mutation just upstream of the polymerase domain of the protein. In addition, each affected child was heterozygous for the G1681A mutation in exon 7 that led to an Ala467Thr substitution in POLG, within the linker region of the protein. PMID- 15122712 TI - SIMPLE mutation in demyelinating neuropathy and distribution in sciatic nerve. AB - Charcot-Marie-Tooth neuropathy type 1C (CMT1C) is an autosomal dominant demyelinating peripheral neuropathy caused by missense mutations in the small integral membrane protein of lysosome/late endosome (SIMPLE) gene. To investigate the prevalence of SIMPLE mutations, we screened a cohort of 152 probands with various types of demyelinating or axonal and pure motor or sensory inherited neuropathies. SIMPLE mutations were found only in CMT1 patients, including one G112S and one W116G missense mutations. A novel I74I polymorphism was identified, yet no splicing defect of SIMPLE is likely. Haplotype analysis of STR markers and intragenic SNPs linked to the gene demonstrated that families with the same mutation are unlikely to be related. The clustering of the G112S, T115N, and W116G mutations within five amino acids suggests this domain may be critical to peripheral nerve myelination. Electrophysiological studies showed that CMT1C patients from six pedigrees (n = 38) had reduced nerve conduction velocities ranging from 7.5 to 27.0m/sec (peroneal). Two patients had temporal dispersion of nerve conduction and irregularity of conduction slowing, which is unusual for CMT1 patients. We report the expression of SIMPLE in various cell types of the sciatic nerve, including Schwann cells, the affected cell type in CMT1C. PMID- 15122713 TI - Developmental effects of von Hippel-Lindau gene deficiency. AB - The histogenetic origin and the basis of the distribution of central nervous system (CNS) hemangioblastomas in the von Hippel-Lindau (VHL) tumor suppressor gene syndrome, VHL disease, are unknown. To better understand hemangioblastoma histogenesis, we analyzed postmortem CNS tissues from four patients with well established diagnosis of VHL disease including development of characteristic tumors and positive family history. Numerous angiomesenchymal tumorlets, which resembled hemangioblastoma, but which also consistently showed distinct histological features, were distributed in the nerve roots, spinal cord, and cerebellum. Genetic analysis consistently showed deletion of the wild-type VHL allele in these tumorlets. Most angiomesenchymal tumorlets were in the dorsal nerve roots; the anterior roots and cerebellum were less frequently affected. Tumorlet distribution was highly consistent in the four cases. In analogy to the wide morphological spectrum of lesions known to exist in VHL kidneys, nerve roots appear to harbor more wide-spread and morphologically heterogeneous changes than previously appreciated. The abundance of tumorlets, associated with highly consistent morphology and topography, suggests a developmental origin of hemangioblastoma. Therefore, in VHL disease, inactivation of the VHL wild-type allele appears necessary, but not sufficient, for the formation of tumor that produces symptoms and neurological disability. PMID- 15122714 TI - Valproate therapy and nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease was defined recently as another symptom of insulin resistance. Continuous therapy with valproate can result in increased body weight and insulin resistance, but no data are yet available on a possible relationship between valproate and nonalcoholic fatty liver disease. We here demonstrate in abdominal ultrasound investigations that characteristics of fatty liver disease were present in 61% of valproate-treated patients as compared with 23% receiving carbamazepine therapy. PMID- 15122715 TI - Clinical impact of antibody formation to botulinum toxin A in children. AB - We studied the clinical impact of neutralizing antibodies to botulinum toxin A that occurred during long-term treatment of children between 1993 and 2001. Antibodies were found in high titers in 35 of 110 (31.8%) samples from individual patients. Antibody formation correlated with secondary nonresponse (p < 0.001). The most significant risk factors for antibody formation were the frequency of treatments (p = 0.0001) and the injection of a higher weight-adapted maximum dose per treatment (p = 0.001). PMID- 15122716 TI - Changes in brain anatomy in focal hand dystonia. AB - No consistent cerebral anatomical abnormality has ever been reported in primary focal hand dystonia (FHD). The present voxel-based morphometry study showed a significant bilateral increase in gray matter in the hand representation area of primary somatosensory and, to a lesser extent, primary motor cortices in 36 patients with unilateral FHD compared with 36 controls. The presence of anatomical changes in the perirolandic cortex for the unaffected hand as well as that for the affected hand suggests that these disturbances may be, at least in part, primary. PMID- 15122717 TI - Inhibition of nonsense-mediated mRNA decay rescues the phenotype in Ullrich's disease. AB - Nonsense-mediated mRNA decay (NMD) is an mRNA surveillance system that eliminates aberrant mRNAs containing premature translation termination codons (PTCs). We evaluated the role of NMD in of Ullrich's disease. The patient has a frameshift mutation with a PTC in the collagen VI alpha2 gene causing the loss of collagen VI and functional defects in extracellular matrix (ECM). The pharmacological block of NMD caused upregulation of the mutant collagen VI alpha2 subunit, resulting in collagen VI assembly and partially functional ECM formation. Our results suggest that NMD inhibitors can be used as a therapeutic tool to rescue some human genetic diseases exacerbated by NMD. PMID- 15122718 TI - Basal ganglia activity remains elevated after movement in focal hand dystonia. AB - Although previous studies of focal hand dystonia have detected cortical sensorimotor abnormalities, little is known about the role of the basal ganglia in this disorder. We report here that when focal hand dystonic patients performed finger-tapping tasks, functional magnetic resonance imaging showed persisting elevations of basal ganglia activity after the tasks ended. We posit that inhibitory control of the basal ganglia may be faulty in focal hand dystonia, and that the increases we observe in "resting" activity may mask basal ganglia abnormalities in standard imaging contrast analyses. PMID- 15122719 TI - The role of human left superior parietal lobule in body part localization. AB - Electrophysiological data in primates suggest that the superior parietal lobule integrates the position of the limbs to construct complex representations of postures. Although in humans the neural basis of these mechanisms remains largely unknown, neuropsychological studies have implicated left superior parietal regions. We devised a simple functional magnetic resonance imaging paradigm aimed at exploring this hypothesis in healthy humans. Strong activation was obtained within the left but not the right superior parietal lobule, providing additional evidence that this structure may play a key role in body part localization processing. PMID- 15122720 TI - Molecular epidemiology of spinocerebellar ataxia type 6. AB - We performed a population-based clinical and molecular genetic study of spinocerebellar ataxia type 6 (SCA6) in the northeast of England. The minimum point prevalence of SCA6 was 1.59 in 100,000 (95% confidence interval [CI], 1.04 2.14), and the number of individuals who either had SCA6 or are at risk of developing SCA6 was at least 5.21 in 100,000 (95% CI, 4.31-6.10), or 1 in 19,210. Microsatellite analysis of the CACNA1A gene indicated a founder effect for SCA6 within this region. PMID- 15122721 TI - Minocycline reduces gadolinium-enhancing magnetic resonance imaging lesions in multiple sclerosis. PMID- 15122722 TI - CCM2 mutations account for 13% of cases in a large collection of kindreds with hereditary cavernous malformations. PMID- 15122726 TI - The statistician and the data monitoring committee. PMID- 15122727 TI - Should statisticians reporting to data monitoring committees be independent of the trial sponsor and leadership? AB - It has long been a fundamental principle of clinical trials that interim comparative data should be kept confidential, with such data accessible only to a small number of individuals responsible for its analysis and monitoring. The rationale for keeping investigators and sponsors blinded to interim data has been extensively discussed, but the possible conflicts of interest that could arise for the statistician who performs the analysis of the interim data and presents it to a data monitoring committee has received little attention. We describe these potential conflicts, and the advantages and disadvantages of approaches that might be taken to minimize them. We have invited commentary on this issue from several statisticians with substantial experience in clinical trials and interim data monitoring. PMID- 15122728 TI - What is the appropriate role of the trial statistician in preparing and presenting interim findings to an independent Data Monitoring Committee in the U.S. Cancer Cooperative Group setting? AB - A recent draft FDA Guidance on the establishment and operation of clinical trial Data Monitoring Committees (DMCs) suggests that statisticians who prepare and present interim analyses to the DMC should be external to the trial sponsor. In the context of the National Cancer Institute (NCI) Cooperative Group program, this recommendation appears to imply that Group study statisticians should be blinded to interim data and excluded from the interim monitoring process in all Cooperative Group trials. In this commentary, it is argued that the benefits of such a policy would be minimal in the Cooperative Group setting and are far outweighed by the tangible benefits of including the study statistician in the monitoring process. PMID- 15122729 TI - The independent statistician for data monitoring committees. AB - Clinical trials are an essential part of the clinical research process. Recently, independent Data Monitoring Committees (DMCs) have been widely implemented to provide scientific and ethical oversight of pivotal clinical trials having irreversible outcomes such as death, stroke, disease recurrence or a serious adverse event. To carry out their responsibility, the DMC reviews interim analyses of accumulating data. We address the motivation for having the preparation and presentation of these interim analyses be conducted by an independent statistician who is not a member of the DMC and who is not the trial's lead or steering committee statistician. These views are based on having served as members of many DMCs as well as having been the independent statistician for several trials. PMID- 15122730 TI - Conflicts of interest in data monitoring of industry versus publicly financed clinical trials. AB - The FDA Guidance, while highly appropriate for industry sponsored trials, need not be imposed on publicly (e.g. NIH) financed clinical trials. While the potential for conflicts of interest exist in the latter, they are in general manageable and pose an acceptable low risk of threatening the integrity of a study. However, the Guidance should heighten the awareness of all investigators in public trials to the potential for the appearance of conflicts and to the need for full and open participation in the management of dualities of interest. PMID- 15122731 TI - Playing safe and preserving integrity: making the FDA model work. AB - The proposed FDA model for Data Monitoring Committees (DMC) includes the recommendation to separate the reporting statistician from the co-ordinating centre statistician. The former writes reports for the DMC; the latter is the statistician responsible for the statistical design of the trial, its ongoing day to-day operation, and the final analysis of the data. This model, while protecting the scientific integrity of the trial, risks producing inadequate reports to the DMC unless the reporting statistician understands the trial and the database. This brief paper describes methods by which the reporting statistician can gain the knowledge necessary to produce informative reports. It also emphasizes that reports produced for the purpose of making decisions during the course of a trial differ in purpose, and should therefore differ in form, from reports summarizing the data and results at the end of the trial. PMID- 15122732 TI - Independence of the statistician who analyses unblinded data. AB - This discussion considers arguments for and against separating responsibility for the unblinded interim analysis of a clinical trial from responsibility for trial management and modifications to the ongoing trial. The degree to which one or different statisticians carry out these responsibilities and thus the degree of statistician independence for the two activities can vary, but a sponsor should recognize that giving a single statistician both responsibilities might limit flexibility in managing the trial, particularly with respect to modifying an ongoing trial. PMID- 15122733 TI - The role of the unblinded sponsor statistician. AB - In clinical trials sponsored by Merck the unblinded statistician is typically an employee of the company. From a pharmaceutical industry perspective, advantages of this approach include ensuring that the unblinded statistician is knowledgeable regarding the experimental treatments, therapeutic area and study objectives; that the quality of the analysis conforms to rigorous standards; and that the allocation schedule, database and interim results are kept confidential. These advantages are felt to outweigh any potential disadvantages. PMID- 15122734 TI - A major trial needs three statisticians: why, how and who? AB - The statistical and scientific integrity of a major clinical trial is enhanced by having three distinct statistician roles: the Study Statistician, the Data Monitoring Committee (DMC) Statistician and the Independent Statistician. In any specific trial, careful attention should be given for selecting the right people to perform these tasks effectively. It is important that there are good communications amongst all three statisticians (and between each statistician and the other trialists) while preserving all confidentiality as regards interim results. Specifically regarding the Independent Statistician, it seems appropriate that they be truly independent having no other trial involvement than producing interim reports that inform the DMC (and be not employed by a commercial sponsor) but at the same time be fully aware of the trial protocol, objectives, organization and database. PMID- 15122735 TI - Test of treatment effect in pre-drug and post-drug count data with zero inflation. AB - A procedure for testing for treatment effect in data similar to the data on premature ventricular contractions (PVC) is presented. We consider a zero inflated beta-binomial model. Based on this model, we develop score tests to test for treatment effect in the data in which observations in the form of counts are recorded before and after applying a therapy. Results of a small simulation experiment, to study small sample behaviour of a score test and a likelihood ratio test, are reported and the PVC data are analysed. Both the score and the likelihood ratio tests show good level properties. Either the score tests or the likelihood ratio tests can be used for testing the presence of treatment effect. The score tests, however, may be preferable because they use estimates of the parameters only under the null hypothesis and in the important range pi<0.5 power of the score test statistic S1 is slightly better than the likelihood ratio statistic LR1. PMID- 15122736 TI - Quantifying and comparing the accuracy of binary biomarkers when predicting a failure time outcome. AB - The positive and negative predictive values are standard measures used to quantify the predictive accuracy of binary biomarkers when the outcome being predicted is also binary. When the biomarkers are instead being used to predict a failure time outcome, there is no standard way of quantifying predictive accuracy. We propose a natural extension of the traditional predictive values to accommodate censored survival data. We discuss not only quantifying predictive accuracy using these extended predictive values, but also rigorously comparing the accuracy of two biomarkers in terms of their predictive values. Using a marginal regression framework, we describe how to estimate differences in predictive accuracy and how to test whether the observed difference is statistically significant. PMID- 15122737 TI - Does the Prentice criterion validate surrogate endpoints? AB - Randomized Phase II or Phase III clinical trials that are powered based on clinical endpoints, such as survival time, may be prohibitively expensive, in terms of both the time required for their completion and the number of patients required. As such, surrogate endpoints, such as objective tumour response or markers including prostate specific antigen or CA-125, have gained widespread popularity in clinical trials. If an improvement in a surrogate endpoint does not itself confer patient benefit, then consideration must be given to the extent to which improvement in a surrogate endpoint implies improvement in the true clinical endpoint of interest. That this is not a trivial issue is demonstrated by the results of an NIH-sponsored trial of anti-arrhythmic drugs, in which the ability to correct an irregular heart beat not only did not correspond to a survival benefit but in fact led to excess mortality. One approach to the validation of surrogate endpoints involves ensuring that a valid between-group analysis of the surrogate endpoint constitutes also a valid analysis of the true clinical endpoint. The Prentice criterion is a set of conditions that essentially specify the conditional independence of the impact of treatment on the true endpoint, given the surrogate endpoint. It is shown that this criterion alone ensures that an observed effect of the treatment on the true endpoint implies a treatment effect also on the surrogate endpoint, but contrary to popular belief, it does not ensure the converse, specifically that the observation of a significant treatment effect on the surrogate endpoint can be used to infer a treatment effect on the true endpoint. PMID- 15122738 TI - Combining several ordinal measures in clinical studies. AB - In medical research, it is rare that a single variable is sufficient to represent all relevant aspects of epidemiological risk, genomic activity, adverse events, or clinical response. Since biological systems tend to be neither linear, nor hierarchical in nature, the assumptions of traditional multivariate statistical methods based on the linear model can often not be justified on theoretical grounds. Establishing concept validity through empirical validation is not only problematic, but also time consuming. This paper proposes the use of u-statistics for scoring multivariate ordinal data and a family of simple non-parametric tests for analysis. The scoring method is demonstrated to be applicable to scoring clinical response profiles in the treatment of psoriasis and then to identifying genomic pathways that best correlate with these profiles. PMID- 15122739 TI - A risk-adjusted Sets method for monitoring adverse medical outcomes. AB - The Sets method has been advocated in previous work as a method for monitoring adverse medical outcomes where the adverse event rate is low. Here, a risk adjusted version of the refined Sets method is presented and an example is given to demonstrate its advantage over the unadjusted method. The method is suitable for any risk distribution and does not assume that changes in risk will be small. A graphical representation, referred to as the Grass plot, of the original and risk-adjusted methods is also given. PMID- 15122740 TI - Methods for assessing reliability and validity for a measurement tool: a case study and critique using the WHO haemoglobin colour scale. AB - Before introducing a new measurement tool it is necessary to evaluate its performance. Several statistical methods have been developed, or used, to evaluate the reliability and validity of a new assessment method in such circumstances. In this paper we review some commonly used methods. Data from a study that was conducted to evaluate the usefulness of a specific measurement tool (the WHO Colour Scale) is then used to illustrate the application of these methods. The WHO Colour Scale was developed under the auspices of the WHO to provide a simple portable and reliable method of detecting anaemia. This Colour Scale is a discrete interval scale, whereas the actual haemoglobin values it is used to estimate are on a continuous interval scale and can be measured accurately using electrical laboratory equipment. The methods we consider are: linear regression, correlation coefficients, paired t-tests plotting differences against mean values and deriving limits of agreement; kappa and weighted kappa statistics, sensitivity and specificity, an intraclass correlation coefficient and the repeatability coefficient. We note that although the definition and properties of each of these methods is well established inappropriate methods continue to be used in medical literature for assessing reliability and validity, as evidenced in the context of the evaluation of the WHO Colour Scale. PMID- 15122741 TI - Generalized log-rank test for mixed interval-censored failure time data. AB - This paper considers the problem of non-parametric treatment comparisons when mixed interval-censored failure time data are available, which often occurs in clinical trials and epidemiological studies. By mixed interval-censored data, we mean that the survival time of interest is observed to belong to an interval or to be right-censored. For the problem, we generalize the most commonly used log rank test for right-censored survival data. Numerical studies are conducted and reported to evaluate and compare the proposed test with the existing method, which indicate that the presented method works well. We apply the method to a data set arising from an AIDS cohort study, that motivated the study. PMID- 15122742 TI - Presentation of multivariate data for clinical use: The Framingham Study risk score functions. AB - The Framingham Heart Study has been a leader in the development and dissemination of multivariable statistical models to estimate the risk of coronary heart disease. These models quantify the impact of measurable and modifiable risk factors on the development of coronary heart disease and can be used to generate estimates of risk of coronary heart disease over a predetermined period, for example the next 10 years. We developed a system, which we call a points system, for making these complex statistical models useful to practitioners. The system is easy to use, it does not require a calculator or computer and it simplifies the estimation of risk based on complex statistical models. This system represents an effort to make available a tool for clinicians to aid in their decision-making process regarding treatment and to assist them in motivating patients toward healthy behaviours. The system is also readily available to patients who can easily estimate their own coronary heart disease risk and monitor this risk over time. PMID- 15122744 TI - Angiogenesis in chronic inflammatory liver disease. AB - Intrahepatic hypoxia may occur during the inflammatory and fibrotic processes that characterize several chronic liver diseases of viral and autoimmune origin. As a consequence, new vascular structures are formed to provide oxygen and nutrients. Angiogenesis involves a tightly regulated network of cellular and molecular mechanisms that result in the formation of functional vessels. Of particular importance are growth factors, molecules involved in matrix remodeling and cell migration, and vessel maturation-related factors. In recent years, a number of studies have examined the expression and function of many pro- and antiangiogenic molecules in the setting of nontumoral chronic liver diseases and liver regeneration. This review examines the potential pathogenetic role of angiogenesis in the context of viral hepatitis, cirrhosis, autoimmune hepatitis, primary biliary cirrhosis, and alcoholic liver disease. The future perspectives for research in this field are outlined. PMID- 15122745 TI - News from Taiwan. PMID- 15122746 TI - Gut flora-based therapy in liver disease? The liver cares about the gut. PMID- 15122747 TI - Whom? When? How? Another piece of evidence for early treatment of acute hepatitis C. PMID- 15122748 TI - Screening in liver disease: report of an AASLD clinical workshop. AB - This report summarizes an AASLD Clinical Workshop that was presented at Digestive Diseases Week 2003 on screening in liver diseases. As newer diagnostic tests become available, many liver diseases and complications of liver disease can be detected at an early asymptomatic stage. In many cases, early detection can lead to earlier treatment and an improved outcome. However, screening for liver diseases in asymptomatic persons has the potential for adverse consequences, including discrimination and stigmatization. The cost of screening programs is significant, and access to screening tests varies in different countries. Future screening programs require careful planning and implementation to balance the benefits, risks, and cost-effectiveness. This review outlines the concepts of screening and their application to a broad range of liver diseases. PMID- 15122749 TI - Short-term interferon-alfa therapy for acute hepatitis C: a randomized controlled trial. AB - Acute hepatitis C often progresses to chronic infection. We undertook a randomized controlled trial to determine whether short-term therapy with interferon (IFN) during acute hepatitis C is effective in preventing the development of chronic hepatitis. Thirty patients with acute hepatitis C were randomized into 1 of 2 treatment groups. IFN therapy was initiated 8 weeks after the onset of acute hepatitis in the early-intervention group and after 1 year of observation in the late-intervention group. Short-term therapy consisted of natural IFN-alfa (6 million units) administered on consecutive days for a period of 4 weeks. Any signs of recrudescence of disease were immediately followed by interval IFN therapy (3 times weekly for 20 weeks). In the early-intervention group, short-term therapy was associated with a sustained virological response in 13 of 15 patients (87%). Follow-up treatment was associated with a sustained virological response in both of the remaining 2 patients (100%). The sustained virological response rate was significantly higher in the early-intervention group (87%, 13 of 15 patients after short-term therapy alone, and 100%, 15 of 15 patients after short-term with or without follow-up therapy) than in the late intervention group (40%, 6 of 15 patients after short-term therapy alone, and 53%, 8 of 15 patients after short-term therapy with or without follow-up therapy, P =.021 and P =.006, respectively). In conclusion, short-term (4 weeks) IFN treatment of patients with acute hepatitis C may be associated with satisfactory results, if initiated at an early stage of the disease. PMID- 15122750 TI - Expression of the CXCR3 ligand I-TAC by hepatocytes in chronic hepatitis C and its correlation with hepatic inflammation. AB - The factors that regulate lymphocyte traffic in chronic hepatitis C (CHC) are not completely defined. Interferon (IFN)-inducible T cell alpha chemoattractant (I TAC) is a relatively new member of the CXCR3 chemokine ligand family that selectively recruits activated T cells to sites of inflammation. To determine if I-TAC plays a role in CHC, we investigated I-TAC expression in hepatitis C virus (HCV)-infected liver biopsy material. I-TAC messenger RNA (mRNA) levels were significantly increased in HCV-infected liver compared with normal liver, which correlated with both portal and lobular inflammation. I-TAC expression was localized to hepatocytes throughout the liver lobule, with those in close proximity to active areas of inflammation expressing the highest concentration of I-TAC. In vitro, I-TAC mRNA and protein expression was inducible in Huh-7 cells following either IFN-alpha or -gamma stimulation and synergistically with tumor necrosis factor (TNF)-alpha. Furthermore, transfection of Huh-7 cells with either poly(I:C) or HCV RNA representing the HCV subgenomic replicon induced I-TAC mRNA expression. HCV replication was also found to modulate I-TAC expression, with stimulation of Huh-7 cells harboring either the HCV subgenomic or genomic replicon showing significantly increased synergistic effects compared with those previously seen in Huh-7 cells alone with IFN-gamma and TNF-alpha. In conclusion, these results suggest I-TAC, one of the most potent chemoattractants for activated T cells, is produced by hepatocytes in the HCV-infected liver and plays an important role in T cell recruitment and ultimately the pathogenesis of CHC. PMID- 15122751 TI - Steatosis and liver cell apoptosis in chronic hepatitis C: a mechanism for increased liver injury. AB - Steatosis is increasingly recognized as a cofactor influencing the progression of fibrosis in chronic hepatitis C; however, the mechanisms by which it contributes to liver injury remain uncertain. We studied 125 patients with chronic hepatitis C to assess the effect of steatosis on liver cell apoptosis and the expression of Bcl-2, Bcl-x(L), Bax, and tumor necrosis factor alpha (TNF-alpha) and the relationship between liver cell apoptosis and disease severity. A significant increase in liver cell apoptosis was seen in liver sections with increasing grade of steatosis (r = 0.42; P <.0001). Hepatic steatosis and previous heavy alcohol consumption were the only two variables independently associated with the apoptotic index. Increasing steatosis was associated with decreased Bcl-2 mRNA levels and an increase in the proapoptotic Bax/Bcl-2 ratio (r = -0.32, P =.007; and r = 0.27, P =.02, respectively). In the absence of steatosis, increased liver cell apoptosis was not associated with stellate cell activation or fibrosis (r = 0.26, P =.11; r = 0.06, P =.71, respectively). In contrast, in the presence of steatosis, increasing apoptosis was associated with activation of stellate cells and increased stage of fibrosis (r = 0.35, P =.047; r = 0.33, P =.03, respectively), supporting the premise that the steatotic liver is more vulnerable to liver injury. In patients with hepatitis C virus genotype 3, there was a significant correlation between TNF-alpha mRNA levels and active caspase-3 (r = 0.54, P =.007). In conclusion, these observations suggest a mechanism whereby steatosis contributes to the progression of liver injury in chronic hepatitis C. Further investigation will be required to determine the molecular pathways responsible for the proapoptotic effect of steatosis and whether this increase in apoptosis contributes directly to fibrogenesis. PMID- 15122752 TI - Improved prediction of fibrosis in chronic hepatitis C using measures of insulin resistance in a probability index. AB - We sought to develop a clinically useful index comprising standard and physiologically relevant variables to predict the probability of significant hepatic fibrosis in subjects with chronic hepatitis C virus (HCV) infection. Fibrosis was graded as mild (stages F0 or F1) or significant (stages F2-F4). Thirty-five clinical and laboratory parameters were analyzed initially in 176 patients with detectable HCV RNA to derive a fibrosis probability index (FPI) to predict significant fibrosis. This index then was validated in a second group of 126 subjects. Among 18 variables associated with severe fibrosis on univariate analysis, multiple logistic regression analysis identified age, aspartate aminotransferase (AST), total cholesterol level, insulin resistance (by homeostasis model), and past alcohol intake as independent predictors of significant fibrosis. The area under the receiver operating characteristic (ROC) curves was 0.84 for the initial cohort and 0.77 for the validation cohort. In the initial cohort, the sensitivity of the FPI based on these five predictors was 96%, and the negative predictive value was 93% at a score of >/=0.2. At scores >/=0.8, the FPI was 94% specific and had a positive predictive value of 87%. In conclusion, an FPI using routinely assessed markers and incorporating a measure of insulin resistance can reliably predict the probability of significant hepatic fibrosis in most patients with chronic HCV infection. Such an index should prove useful to guide decision making regarding the need for liver biopsy, and potentially for avoiding or deferring biopsy in a large proportion of patients with mild liver disease. PMID- 15122753 TI - Low membrane protein sulfhydrils but not G6PD deficiency predict ribavirin induced hemolysis in hepatitis C. AB - Hemolysis is a frequent adverse effect of ribavirin (RBV). It has been suggested that oxidative stress plays a role, but mechanisms and predictive risk factors for severe forms remain unknown. Markers of redox status were determined in erythrocytes of 34 patients with hepatitis C-four of them with glucose-6 phosphate-dehydrogenase (G6PD) deficiency-before and during treatment with RBV and interferon (IFN) and were compared with 10 healthy control subjects. In addition, erythrocytes were incubated with RBV, and the effects of dipyridamole (DPD), diethylmaleate (DEM), and glutathione ester (GSHE) were studied in vitro. Of the 30 patients without G6PD deficiency who were treated with RBV and IFN alpha, five developed major hemolysis (Delta hemoglobin > 6 g/dL) and 25 developed minor hemolysis (Delta hemoglobin < 2.5 g/dL). Patients with major hemolysis had lower median pretreatment values of membrane protein sulfhydrils than patients with minor hemolysis (28.4 vs. 36.7 nmol/mg, P <.001). Erythrocytes of G6PD-deficient patients were not more susceptible to RBV-induced hemolysis. In in vitro incubations of erythrocytes, DEM enhanced the RBV-induced decrease of glutathione, protein sulfhydrils, and osmotic resistance. Supplementation of GSHE and DPD prevented the RBV-induced decrease in osmotic resistance, adenosyl triphosphate (ATP), and 2,3-diphosphoglycerate (DPG), the loss of glutathione and protein sulfhydrils, and the formation of thiobarbituric acid reactive substances (TBARs). In conclusion, the data indicate that low membrane protein sulfhydrils prior to therapy but not G6PD deficiency are predictive of RBV-induced major hemolysis. In vitro, GSHE and DPD reduce the RBV-associated oxidative stress in erythrocytes and prevent the increase in osmotic fragility, suggesting that these compounds might decrease the risk of hemolysis in patients. PMID- 15122754 TI - Recently primed CD8+ T cells entering the liver induce hepatocytes to interact with naive CD8+ T cells in the mouse. AB - Large number of T cells traffic through the liver. In order to examine the effects of such traffic on the phenotype of hepatocytes, we vaccinated mice using DNA vaccines encoding antigens with MHC class I-binding epitopes. Small numbers of activated CD8(+) T blasts (10(5)-10(6)/liver) changed the surface phenotype and cytokine expression profile of hepatocytes (HCs). HCs upregulate surface expression of major histocompatibility complex (MHC) class I molecules and CD1d but not MHC class II molecules Qa-1, CD80, CD86, CD54, or CD95; in addition, they expressed/secreted interleukin (IL)-10 and IL-4 but not IL-1, IL-6, IL-13, interferon (IFN)-gamma, tumor necrosis factor (TNF), IL-4, or IL-27 (i.e., they acquire the HC* phenotype). HCs* (but not HCs) induced specific activation, proliferation, and IFN-gamma, TNF, and IL-13 release of cocultured naive CD8(+) T cells. In contrast to the specific activation of naive CD8(+) T cells by dendritic cells (DCs), specific CD8(+) T cell activation by HC* was not down modulated by IFN-alphabeta. Only recently activated CD8(+) T blasts (but not recently activated CD4(+) T blasts or activated cells of the innate immune system, including natural killer T [NKT] cells) induced the HC* phenotype that is prominent from day 10 to day 20 postvaccination (i.e., time points at which peak numbers of recently primed CD8(+) T blasts are found in the liver). In conclusion, recently activated CD8(+) T blasts that enter the liver postimmunization in small numbers can transiently modulate the phenotype of HC, allowing them to activate naive CD8(+) T cells with unrelated specificities. PMID- 15122755 TI - Activation of hepatic Nrf2 in vivo by acetaminophen in CD-1 mice. AB - The transcription factor NF-E2-related factor 2 (Nrf2) plays an essential role in the mammalian response to chemical and oxidative stress through induction of hepatic phase II detoxification enzymes and regulation of glutathione (GSH). Enhanced liver damage in Nrf2-deficient mice treated with acetaminophen suggests a critical role for Nrf2; however, direct evidence for Nrf2 activation following acetaminophen exposure was previously lacking. We show that acetaminophen can initiate nuclear translocation of Nrf2 in vivo, with maximum levels reached after 1 hour, in a dose dependent manner, at doses below those causing overt liver damage. Furthermore, Nrf2 was shown to be functionally active, as assessed by the induction of epoxide hydrolase, heme oxygenase-1, and glutamate cysteine ligase gene expression. Increased nuclear Nrf2 was found to be associated with depletion of hepatic GSH. Activation of Nrf2 is considered to involve dissociation from a cytoplasmic inhibitor, Kelch-like ECH-associated protein 1 (Keap1), through a redox-sensitive mechanism involving either GSH depletion or direct chemical interaction through Michael addition. To investigate acetaminophen-induced Nrf2 activation we compared the actions of 2 other GSH depleters, diethyl maleate (DEM) and buthionine sulphoximine (BSO), only 1 of which (DEM) can function as a Michael acceptor. For each compound, greater than 60% depletion of GSH was achieved; however, in the case of BSO, this depletion did not cause nuclear translocation of Nrf2. In conclusion, GSH depletion alone is insufficient for Nrf2 activation: a more direct interaction is required, possibly involving chemical modification of Nrf2 or Keap1, which is facilitated by the prior loss of GSH. PMID- 15122756 TI - Pivotal role of superoxide anion and beneficial effect of antioxidant molecules in murine steatohepatitis. AB - Nonalcoholic fatty liver disease, frequently associated with obesity, can lead to nonalcoholic steatohepatitis (NASH) and cirrhosis. The pathophysiology of NASH is poorly understood, and no effective treatment is available. In view of a potential deleterious role for reactive oxygen species (ROS), we investigated the origin of ROS overproduction in NASH. Mitochondrial production of ROS and its alterations in the presence of antioxidant molecules were studied in livers from ob/ob mice that bear a mutation of the leptin gene and develop experimental NASH. N-acetyl-cysteine and the superoxide dismutase (SOD) mimics ambroxol, manganese [III] tetrakis (5,10,15,20 benzoic acid) (MnTBAP), and copper [II] diisopropyl salicylate (CuDIPS) were used to target different checkpoints of the oxidative cascade to determine the pathways involved in ROS production. Liver mitochondria from ob/ob mice generated more O(2)*- than those of lean littermates (P <.01). Ex vivo, all three SOD mimics decreased O(2)*- generation (P <.001) and totally inhibited lipid peroxidation (P <.001) versus untreated ob/ob mice. Those modifications were associated with in vivo improvements: MnTBAP and CuDIPS reduced weight (P <.02) and limited the extension of histological liver steatosis by 30% and 52%, respectively, versus untreated ob/ob mice. In conclusion, these data demonstrate deleterious effects of superoxide anions in NASH and point at the potential interest of nonpeptidyl mimics of SOD in the treatment of NASH in humans. PMID- 15122757 TI - Administration of the potent PPARalpha agonist, Wy-14,643, reverses nutritional fibrosis and steatohepatitis in mice. AB - Administration of a methionine and choline deficient (MCD) diet to rodents causes progressive fibrosing steatohepatitis pathologically similar to human metabolic steatohepatitis. We have previously shown that the peroxisome proliferator activated receptor-alpha (PPARalpha) agonist, Wy-14,643, prevented the development of MCD diet-induced steatohepatitis. We have now tested whether Wy 14,643 ameliorates established steatohepatitis and fibrosis. Male C57BL6 mice were fed the MCD diet for 51 days to induce severe steatohepatitis. They were then treated with Wy-14,643 together with the MCD diet for 5 or 12 days; positive controls continued on the MCD diet for 5 or 12 days. After 5 days of Wy-14,643 treatment, alanine aminotransferase (ALT) levels were significantly decreased, steatohepatitis less severe, and hepatic lipoperoxides significantly reduced. After 12 days, hepatic triglycerides were normalized and there was near resolution of histological changes. MCD dietary feeding was associated with increased expression of vascular cell adhesion molecule (VCAM)-1, and increased numbers of activated macrophages in the liver. Treatment with Wy-14,643 reduced VCAM-1 expression and macrophage numbers. MCD diet-fed mice developed hepatic fibrosis with increased hepatic collagen alpha1(I), tissue inhibitor of metalloproteinases (TIMP)-1, TIMP-2, and matrix metalloproteinase (MMP)-13 mRNA levels. After treatment with Wy-14,643, expression of these genes was reduced in a manner that paralleled the reduction in activated hepatic stellate cells and near resolution of liver fibrosis. In conclusion, the present study shows that MCD diet-induced fibrosing steatohepatitis can be reversed by treatment with Wy 14,643. It is likely that activation of PPARalpha reverses fibrosis indirectly by reducing stimuli, such as lipid peroxides, and activation of cells responsible for promoting hepatic fibrosis. PMID- 15122758 TI - Murine alanine aminotransferase: cDNA cloning, functional expression, and differential gene regulation in mouse fatty liver. AB - Alanine aminotransferase (ALT) is a widely used index of liver integrity or hepatocellular damage in clinics as well as a key enzyme in intermediatary metabolism. In this study, we have cloned the complementary DNAs of murine homologues of human alanine aminotransferase 1 and 2 (ALT1 and ALT2). The deduced peptides of murine ALT1 (mALT1) and ALT2 (mALT2) share 87% and 93% identity, respectively, with their human counterparts at the amino acid level. Murine ALT genes localize to separate chromosomes, with mALT1 gene (gpt1) on chromosome 15 and mALT2 gene (gpt2) on chromosome 8. The murine gpt1 and gpt2 differ in messenger RNA expression: gpt1 is mainly expressed in liver, bowel, and white adipose tissue and gpt2 is highly expressed in muscle, liver, and white adipose tissue. Expression of recombinant mALT1 and mALT2 proteins in Escherichia coli (E. coli) produced functional enzymes that catalyze alanine transamination. The potential diagnostic value of ALT isoenzymes in liver disease was evaluated in an obese animal model. In fatty livers of obese mice, ALT2 gene expression is induced 2-fold, but ALT1 remains the same. Furthermore, in fatty liver, total hepatic ALT activity is elevated significantly by 30% whereas aspartate aminotransferase (AST) activity remains unchanged. In conclusion, these results indicate that ALT2 may be responsible for the increased ALT activity in hepatic steatosis and provide evidence that an ALT isoenzyme-specific assay may have more diagnostic value than the total ALT activity assay currently in clinical use. PMID- 15122759 TI - Hepatic transmethylation reactions in micropigs with alcoholic liver disease. AB - Alcoholic liver disease is associated with abnormal hepatic methionine metabolism, including increased levels of homocysteine and S-adenosylhomocysteine (SAH) and reduced levels of S-adenosylmethionine (SAM) and glutathione (GSH). The concept that abnormal methionine metabolism is involved in the pathogenesis of alcoholic liver disease was strengthened by our previous findings in a micropig model where combining dietary folate deficiency with chronic ethanol feeding produced maximal changes in these metabolites together with early onset of microscopic steatohepatitis and an eightfold increase in plasma aspartate aminotransferase. The goal of the present study was to determine potential mechanisms for abnormal levels of these methionine metabolites by analyzing the transcripts and activities of transmethylation enzymes in the livers of the same micropigs. Ethanol feeding or folate deficiency, separately or in combination, decreased transcript levels of methylenetetrahydrofolate reductase (MTHFR), methionine adenosyltransferase (MAT1A), glycine-N-methyltransferase (GNMT) and S adenosylhomocysteine hydrolase (SAHH). Ethanol feeding alone reduced the activities of methionine synthase (MS) and MATIII and increased the activity of GNMT. Each diet, separately or in combination, decreased the activities of MTHFR and SAHH. In conclusion, the observed abnormal levels of methionine metabolites in this animal model of accelerated alcoholic liver injury can be ascribed to specific effects of ethanol with or without folate deficiency on the expressions and activities of hepatic enzymes that regulate transmethylation reactions. These novel effects on transmethylation reactions may be implicated in the pathogenesis of alcoholic liver disease. PMID- 15122760 TI - Topoisomerase inhibitor camptothecin sensitizes mouse hepatocytes in vitro and in vivo to TNF-mediated apoptosis. AB - Topoisomerases are nuclear enzymes that maintain and modulate DNA structure. Inhibitors of topoisomerases like camptothecin (CPT), etoposide, and others are widely used antitumor drugs that interfere with transcription, induce DNA strand breaks, and trigger apoptosis preferentially in dividing cells. Because transcription inhibitors (actinomycin D, galactosamine, alpha-amanitin) sensitize primary hepatocytes to the cytotoxic action of tumor necrosis factor (TNF), we reasoned whether topoisomerase inhibitors would act similarly. CPT alone was not toxic to primary cultured murine hepatocytes. When incubated with CPT, murine hepatocytes displayed an inhibition of protein synthesis and were thereby rendered sensitive to apoptosis induction by TNF. Apoptosis was characterized by morphology (condensed/fragmented nuclei, membrane blebbing), caspase-3-like protease activity, fragmentation of nuclear DNA, and late cytolysis. Hepatocytes derived from TNF receptor-1 knockout mice were resistant to CPT/TNF-induced apoptosis. CPT treatment completely abrogated the TNF-induced NF-kappa B activation, and mRNA expression of the antiapoptotic factors TNF-receptor associated factor 2, FLICE-inhibitory protein, and X-linked inhibitor of apoptosis protein was also inhibited by CPT. The caspase inhibitors benzyloxycarbonyl-Val-Ala-Asp-(OMe)-fluoromethylketone (zVAD-fmk) and benzyloxycarbonyl-Asp(OMe)-Glu(OMe)-Val-Asp(OMe)-chloromethylketone (zDEVD-fmk), as well as depletion of intracellular ATP by fructose prevented CPT/TNF-induced apoptosis. In vivo, CPT treatment sensitized mice to TNF-induced liver damage. In conclusion, the combination of topoisomerase inhibition and TNF blocks survival signaling and elicits a type of hepatocyte death similar to actinomycin D/TNF or galactosamine/TNF. During antitumor treatment with topoisomerase inhibitors, an impaired immune function often results in opportunistic infections, a situation where the systemic presence of TNF might be critical for the hepatotoxicity reported in clinical topoisomerase inhibitor studies. PMID- 15122761 TI - Liver NK cells expressing TRAIL are toxic against self hepatocytes in mice. AB - Although it is known that activation of natural killer (NK) cells causes liver injury, the mechanisms underlying NK cell-induced killing of self-hepatocytes are not clear. We demonstrated that liver NK cells have cytotoxicity against normal syngeneic hepatocytes in mice. Polyinosinic-polycytidylic acid (poly I:C) treatment enhanced hepatocyte toxicity of liver NK cells but not that of spleen NK cells. Unlike NK cells in other tissues, approximately 30%-40% of liver NK cells constitutively express tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). An in vitro NK cell cytotoxic assay revealed that hepatocyte toxicity of liver NK cells from both naive and poly I:C-treated mice was inhibited partially by an anti-TRAIL monoclonal antibody (mAb) alone and completely by the combination with anti-Fas ligand (FasL) mAb and a perforin inhibitor, concanamycin A, indicating contribution of TRAIL to NK cell-mediated hepatocyte toxicity. The majority of TRAIL(+) NK cells lacked expression of Ly-49 inhibitory receptors recognizing self-major histocompatibility complex class I, indicating a propensity to targeting self-hepatocytes. Poly I:C treatment significantly upregulated the expression of Ly-49 receptors on TRAIL(-) NK cells. This might be a compensatory mechanism to protect self-class I-expressing cells from activated NK cell-mediated killing. However, such compensatory alteration was not seen at all in the TRAIL(+) NK cell fraction. Thus, liver TRAIL(+) NK cells have less capacity for self-recognition, and this might be involved in NK cell-dependent self-hepatocyte toxicity. In conclusion, our findings are consistent with a model in which TRAIL-expressing NK cells play a critical role in self-hepatocyte killing through poor recognition of MHC. PMID- 15122762 TI - Interleukin 22 (IL-22) plays a protective role in T cell-mediated murine hepatitis: IL-22 is a survival factor for hepatocytes via STAT3 activation. AB - The central role of T cell activation in hepatocellular injury has been well documented. In this article, we provide evidence suggesting that T cells may also play a protective role in liver disease by releasing interleukin-22 (IL-22), a recently identified T cell-derived cytokine whose biological significance is unclear. IL-22 messenger RNA and protein expression are significantly elevated in T cell-mediated hepatitis induced by concanavalin A (ConA) but are less extensively elevated in the carbon tetrachloride-induced liver injury model. Activated CD3(+) T cells are likely responsible for the production of IL-22 in the liver after injection of ConA. The IL-22 receptor is normally expressed at high levels by hepatocytes and further induced after ConA injection. IL-22 blockade with a neutralizing antibody reduces signal transducer and activator of transcription factor 3 (STAT3) activation and worsens liver injury in T cell mediated hepatitis, whereas injection of recombinant IL-22 attenuates such injury. In vitro treatment with recombinant IL-22 or overexpression of IL-22 promotes cell growth and survival in human hepatocellular carcinoma HepG2 cells. Stable overexpression of IL-22 in HepG2 cells constitutively activates STAT3 and induces expression of a variety of antiapoptotic (e.g., Bcl-2, Bcl-xL, Mcl-1) and mitogenic (e.g., c-myc, cyclin D1, Rb2, CDK4) proteins. Blocking STAT3 activation abolishes the antiapoptotic and mitogenic actions of IL-22 in hepatic cells. In conclusion, the T cell-derived cytokine IL-22 is a survival factor for hepatocytes; this suggests that T cell activation may also prevent and repair liver injury by releasing hepatoprotective cytokine IL-22 in addition to its previously documented central role in hepatocellular injury. PMID- 15122763 TI - Cytokines increase CRE binding but decrease CRE-mediated reporter activity in rat hepatocytes by increasing c-Jun. AB - The cyclic AMP response element (CRE) has been implicated in the regulation of the expression of many genes and cellular processes important in hepatocyte function. CRE sites exist in the promoter regions of several genes expressed during inflammation. Numerous studies on the role of CRE in hepatocyte gene expression have been performed in resting hepatocytes, but the role of CRE during inflammation is unknown. To evaluate the regulation of CRE-mediated transcription during sepsis, cultured hepatocytes were exposed to proinflammatory cytokines and lipopolysaccharide (LPS) was injected into rats. Nuclear proteins were collected and CRE binding activity measured by electromobility shift assay (EMSA) using a consensus CRE oligonucleotide. CRE binding activity was increased in vitro by cytokines and in vivo by LPS administration but CRE-dependent reporter activity was decreased by cytokine stimulation. A c-jun N-terminal kinase (JNK) inhibitor reversed the cytokine-induced increase in CRE binding and increased CRE-dependent reporter activity. Supershift assays indicated that cyclic AMP response element binding protein (CREB) and c-Jun proteins were included in the CRE binding complex. CREB induced and c-Jun suppressed reporter activity using a CRE dependent construct transfected into cultured primary hepatocytes. In conclusion, these data demonstrate that proinflammatory cytokines regulate CRE binding and activity in cultured hepatocytes and suggest that sepsis-induced changes in CRE binding may participate in the cellular response to inflammation. PMID- 15122764 TI - 2-acetylaminofluorene dose-dependent differentiation of rat oval cells into hepatocytes: confocal and electron microscopic studies. AB - The 2-acetylaminofluorene (AAF)/partial hepatectomy (PH) model is one of the most extensively studied experimental systems for oval cell proliferation and differentiation. We have previously described the oval cells as forming ductular structures surrounded by basement membrane, representing extensions of the canals of Hering. Herein we analyze the differentiation of oval cells into hepatocytes after varying degrees of liver damage induced by AAF. At a low dose of AAF, most oval cells synchronously differentiate into small hepatocytes by 6 days after the PH, resulting in complete restoration of the liver structure in 10 days. Higher doses of AAF delay the differentiation process and the new hepatocytes form foci, in contrast to what is observed at the low dose. Qualitatively, the differentiation process seems to be identical at the cellular level under both conditions. The transition from the expanding oval cell population into hepatocytes was correlated with the upregulation of hepatocyte nuclear factor 4 and the disappearance of the basement membrane. Also, the differentiation of oval cells into hepatocytes coincided with the loss of alpha-fetoprotein and OV-6 staining, and the replacement of the biliary cell-specific alpha6 integrin and connexin 43 with the hepatocyte-specific alpha1 integrin and connexin 32. In addition, bile canaliculi form between the new hepatocytes. In conclusion, these results indicate the rate of oval cell differentiation into hepatocytes is context dependent and suggest that, under favorable conditions, oval cells can complete this process much faster than previously appreciated. PMID- 15122765 TI - Thy1-positive mesenchymal cells promote the maturation of CD49f-positive hepatic progenitor cells in the mouse fetal liver. AB - Previously, we reported a system to enrich mouse fetal hepatic progenitor cells (HPCs) by forming cell aggregates. In this study, we sorted two cell populations, CD49f(+)Thy1(-)CD45(-) cells (CD49f-positive cells) and CD49f(+/-)Thy1(+)CD45(-) cells (Thy1-positive cells), from the cell aggregates using a flow cytometer. CD49f-positive cells stained positive for endodermal specific markers such as alpha-fetoprotein (AFP), albumin (ALB), and cytokeratin 19 (CK19), and are thus thought to be HPCs. However, Thy1-positive cells were a morphologically heterogeneous population; reverse-transcription polymerase chain reaction (RT PCR) and immunocytochemical analyses revealed the expression of mesenchymal cell markers such as alpha-smooth muscle actin, desmin, and vimentin, but not of AFP, ALB, or CK19. Therefore, Thy1-positive cells were thought to be of a mesenchymal lineage. When these two cell populations were co-cultured, the CD49f-positive colonies matured morphologically and stored a significant amount of glycogen. Furthermore, real-time RT-PCR demonstrated an increased expression of tyrosine amino transferase and tryptophan oxygenase mRNA, and transmission electron microscopy confirmed that co-cultured cells produced mature hepatocytes. However, when CD49f-positive cells were cultured alone or when the two populations were cultured separately, the CD49f-positive cells did not mature. These results indicate that CD49f-positive cells are primitive hepatic endodermal cells with the capacity to differentiate into hepatocytes, and that Thy1-positive cells promote the maturation of CD49f-positive cells by direct cell-to-cell contact. In conclusion, we were able to isolate CD49f-positive primitive hepatic endodermal cells and Thy1-positive mesenchymal cells and to demonstrate the requirement of cell-to-cell contact between these cell types for the maturation of the hepatic precursors. PMID- 15122766 TI - An oncolytic adenoviral vector of Smac increases antitumor activity of TRAIL against HCC in human cells and in mice. AB - Hepatocellular carcinoma (HCC) displays a high resistance to tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated cell death. To increase sensitivity of HCC cells to TRAIL, we have constructed an oncolytic adenoviral vector (ZD55) and used this vector to deliver second mitochondria-derived activator of caspases (Smac) and TRAIL genes (ZD55-Smac and ZD55-TRAIL, respectively) into HCC cells. Our data showed that human HCC cells express high levels of inhibitor of apoptosis proteins (IAPs). Transfected HCC cells expressing exogenous X-linked IAPs (XIAPs) displayed more resistance to TRAIL. The expression of Smac led to rapid and potent activation of apoptosis in HCC cells after infection with ZD55-Smac. The activation of caspases and induction of apoptosis could be enhanced further through coinfection with ZD55-TRAIL. The combined treatment of ZD55-Smac and ZD55-TRAIL resulted in significant reduction of XIAP expression levels. In addition, our in vivo data in mice showed only a partial response in the established tumor treated either by ZD55-Smac or ZD55 TRAIL alone. By contrast, complete tumor regression was observed by combination of ZD55-Smac and ZD55-TRAIL in all treated animals. This strong antitumoral activity achieved by this combination was due to a dramatic induction of tumor cell apoptosis in the treated tumors. In conclusion, our data indicate that Smac antagonizes the IAPs in HCC tumor cells and enhances tumor cell death induced by TRAIL in the oncolytic adenoviral vector. The combination of Smac and TRAIL delivered by way of the oncolytic adenoviral vector would provide a useful strategy for therapy of HCC and might also be applied to other IAPs abundant in cancers. PMID- 15122767 TI - Down-regulation of the organic cation transporter 1 of rat liver in obstructive cholestasis. AB - The liver plays a major role in biotransformation and elimination of various therapeutic agents and xenobiotics, many of which are organic cations and substrates of the organic cation transporter 1 (Oct1, Slc22a1). Oct1 is expressed at the basolateral membranes of hepatocytes and proximal renal tubules. Although Oct1 is the major uptake mechanism in hepatocytes for many pharmaceutical compounds, little is known about the effects of liver injury on this process. Our aim was to investigate the effects of obstructive cholestasis on Oct1 expression and function in liver and kidney. The effects of bile duct ligation (BDL) on Oct1 protein, messenger RNA (mRNA) expression, and tissue localization were determined in rat liver and kidney with Western analysis, real-time reverse transcriptase mediated polymerase chain reaction (RT-PCR), and immunofluorescence. To assess Oct1 function, the model substrate tetraethylammonium ([(14)C]TEA) was administered intravenously to BDL and control rats and distribution of radioactivity was determined. Oct1 protein significantly decreased in cholestatic livers to 42.1 +/- 17.7% (P <.001), 15.5 +/- 4.7% (P <.05), and 8.6 +/- 2.7% (P <.05) of controls after 3, 7, and 14 days, respectively, but not in kidneys. Hepatic Oct1 mRNA decreased to 77.2 +/- 12.7%, 40.7 +/- 8.1% (P <.05), and 50.3 +/- 7.5% (P <.05) 3, 7, and 14 days after BDL, respectively. Tissue immunofluorescence corroborated these data. Hepatic accumulation of [(14)C]TEA in 14-day BDL rats was reduced to 29.6 +/- 10.9% of controls (P <.0005). In conclusion, obstructive cholestasis down-regulates Oct1 and impairs Oct1-mediated uptake in rat liver, suggesting that hepatic uptake of small cationic drugs may be impaired in cholestatic liver injury. PMID- 15122768 TI - A double-blind randomized controlled trial of infliximab associated with prednisolone in acute alcoholic hepatitis. AB - Tumor necrosis factor-alpha (TNF-alpha) may contribute to the progression of acute alcoholic hepatitis (AAH). The aim of this study was to evaluate the efficacy of an association of infliximab and prednisolone at reducing the 2-month mortality rate among patients with severe AAH. Patients with severe AAH (Maddrey score >/=32) were randomly assigned to group A receiving intravenous infusions of infliximab (10 mg/kg) in weeks 0, 2, and 4; or group B receiving a placebo at the same times. All patients received prednisolone (40 mg/day) for 28 days. Blood neutrophil functional capacities were monitored over 28 days. After randomization of 36 patients, seven patients from group A and three from group B died within 2 months. The probability of being dead at 2 months was higher (not significant [NS]) in group A (39% +/- 11%) than in group B (18% +/- 9%). The study was stopped by the follow-up committee and the sponsor (Assistance Publique-Hopitaux de Paris). The frequency of severe infections within 2 months was higher in group A than in group B (P <.002). This difference was potentially related to a significantly lower ex vivo stimulation capacity of neutrophils. There were no differences between the two groups in terms of Maddrey scores at any time point. In conclusion, three infusions of 10 mg/kg of infliximab in association with prednisolone may be harmful in patients with severe AAH because of the high prevalence of severe infections. PMID- 15122769 TI - Asialoglycoprotein receptor facilitates hemolysis in patients with alcoholic liver cirrhosis. AB - Hemolysis in patients with advanced alcoholic liver disease is a common clinical problem and indicates an unfavorable prognosis. In many cases, the etiology of the hemolysis remains unknown. We observed three patients with alcoholic liver disease, suffering from severe hemolytic anemia, requiring multiple blood transfusions. Steroid therapy was ineffective and two of the patients died. All patients had a soluble variant of the human asialoglycoprotein receptor (s-ASGP R) in their serum, as well as high titers of autoantibodies against this receptor (anti-ASGP-R). Consecutively, examination of 60 patients with alcoholic liver disease revealed a high incidence for s-ASGP-R (36%) and anti-ASGP-R (27%) in patients with alcoholic liver cirrhosis (ALC) compared to patients with cirrhosis due to viral hepatitis. The potential etiology of hemolysis was studied in vitro on erythrocytes from patients with ALC and from healthy donors. Isolated ASGP-R but not anti-ASGP-R bound to the surface of erythrocytes preferentially of blood group A1 and caused dose-dependent agglutination and hemolysis, while this phenomenon was much lower using erythrocytes of the blood group B and almost absent with blood group O-erythrocytes. Furthermore, agglutination and hemolysis only occurred in erythrocytes from ALC-patients or after the pre-treatment of cells with neuraminidase. ASGP-R induced agglutination and hemolysis was blocked by the competitive ASGP-R inhibitor asialofetuin. In conclusion, our results indicate a new, non-immunological mechanism for hemolysis in patients with alcoholic liver disease, mediated through agglutination by a soluble variant of the human asialoglycoprotein receptor and mechanical shear stress. PMID- 15122770 TI - In vitro comparison of the molecular adsorbent recirculation system (MARS) and single-pass albumin dialysis (SPAD). AB - The detoxification capacities of single-pass albumin dialysis (SPAD), the molecular adsorbents recirculation system, (MARS) and continuous veno-venous hemodiafiltration (CVVHDF) were compared in vitro. In each experiment 4,100 mL of toxin-loaded human plasma was processed for 6.5 hours. MARS treatment (n = 6) was undertaken in combination with CVVHDF. For SPAD (n = 6) and CVVHDF (n = 6) a high flux hollow fiber hemodiafilter (identical to the MARS filter) was used. Levels of ammonia, urea, creatinine, bilirubin, and bile acids were determined. Concentrations before and after application of detoxification procedures were expressed as differences and were compared using the Kruskal-Wallis test. Post hoc comparisons for pairs of groups were adjusted according to Bonferroni-Holm. Time, group, and interaction effects were tested using the nonparametric ANOVA model for repeated measurements. SPAD and CVVHDF induced a significantly greater reduction of ammonia levels than MARS. No significant differences were found among SPAD, MARS, and CVVHDF with respect to other water-soluble substances. SPAD induced a significantly greater reduction in bilirubin levels than MARS. Reductions in bile acid levels were similar for SPAD and MARS. When operating MARS in continuous veno-venous hemodialysis mode, as recommended by the manufacturer, no significant differences in the removal of bilirubin, bile acids, urea, and creatinine were found. However, MARS in continuous veno-venous hemodialysis mode was significantly less efficient in removing ammonia than MARS in CVVHDF mode. In conclusion, the detoxification capacity of SPAD is similar to or even greater than that of MARS. PMID- 15122771 TI - Caspase induction by IgA antimitochondrial antibody: IgA-mediated biliary injury in primary biliary cirrhosis. AB - Anti-mitochondrial antibodies (AMAs) have long been recognized as a serological hallmark of primary biliary cirrhosis (PBC). Although high titers of immunoglobulin (Ig)A AMAs are found in bile, saliva, and urine of patients, a pathogenic role for this antibody has remained elusive. Functional studies of this IgA in general have been impeded by low quantities of antibody and the inability to recover antigen-specific IgA in dimeric form. Using a newly defined synthetic group A. Streptococcus derived peptide, we purified large quantities of dimeric and monomeric IgA from patient sera. The purified IgA was incubated with Madine-Darby canine kidney (MDCK) cells transfected with the human polymeric Ig receptor (pIgR) and the cells studied by flow cytometric analysis for binding of carboxyfluorescein conjugated VAD-fmk peptide to activated caspase enzymes. A total of 87% of PBC patients that were anti-PDC-E2 positive had serum IgA that increased caspase activation in MDCK-pIgR+ cells compared to serum-derived IgA from controls with a maximum reaction 48 hours after addition of IgA. The titer of anti-PDC-E2 IgA among the PBC patients strongly correlated with caspase activation (cc = 0.88). Pre-absorption of the IgA using recombinant 2-oxo-acid dehydrogenase complex significantly diminished this activation. IgG from the same PBC patients did not induce caspase activation. These data suggest that during transcytosis through pIgR-positive cells, exposure to PDC-E2-specific dimeric IgA results in the initiation of caspase activation. In conclusion, we propose that due to an even greater concentration of dimeric IgA in biliary and mucosal secretions, constant transcytosis would render the exposed cells more susceptible to apoptosis resulting in subsequent bile duct damage. PMID- 15122772 TI - The zonal distribution of megamitochondria with crystalline inclusions in nonalcoholic steatohepatitis. AB - Megamitochondria with crystalline inclusions (MMC) have been previously described in nonalcoholic fatty liver; however, their distribution within hepatic zones is unknown. We sought to determine this distribution from the core liver biopsy specimens of 31 patients: 8 males and 23 females, age range 21 to 72 years. Twenty-nine showed evidence of nonalcoholic steatohepatitis (NASH) on biopsy with steatosis, inflammation, varying degree of fibrosis, ballooned hepatocytes, and Mallory hyaline, and two patients had cryptogenic cirrhosis thought to represent "burned out" NASH. Identified by transmission electron microscopy, the abundance of MMC was compared between low-stage (fibrosis stages 1 and 2) and high-stage (fibrosis stages 3 and 4) groups and between zones with or without difference in fibrosis stage. Regardless of stage, the MMC were distributed equally in all zones and were abundant similarly in low- and high-stage groups. This abundance did not correlate with the degree of oxidative stress (4-hydroxynonenal staining) or with the abundance of ballooned hepatocytes. Consistent with age as a risk factor for more severe disease, the median age for the low-stage group was significantly lower than that of the high-stage group (P =.003). In conclusion, in NASH, the MMC seem to be distributed randomly among zones and without variation in abundance, regardless of the fibrosis stage. The exact function of these structures remains to be defined. In this study, their presence did not seem to correlate with the light microscopic injury pattern represented by ballooned hepatocytes or degree of oxidative stress defined by immunostaining for 4-hydroxynonenal. PMID- 15122773 TI - Hepatic adducts, circulating antibodies, and cytokine polymorphisms in patients with diclofenac hepatotoxicity. AB - Diclofenac is a nonsteroidal anti-inflammatory drug that causes rare but serious hepatotoxicity, the mechanism of which is unclear. The purpose of the present study was to explore the potential role played by the immune processes. Antibodies to diclofenac metabolite-modified liver protein adducts were detected in the sera of seven out of seven patients with diclofenac-induced hepatotoxicity, 12 of 20 subjects on diclofenac without hepatotoxicity, and none of four healthy controls. The antibodies recognized adducts expressed in livers from rats treated with multiple doses of diclofenac, but not in those given single doses. In addition, several potential diclofenac adducts were identified in the liver of a patient with diclofenac-induced hepatic failure, but not from a normal human donor liver, by immunoblotting with an adduct-selective rabbit antiserum. To determine whether or not polymorphisms in genes encoding cytokine related proteins influence susceptibility to hepatotoxicity, genotyping for the polymorphisms -627 in the interleukin (IL)-10 gene, -590 in the IL-4 gene, and codon 551 in the IL-4 receptor (IL-4R) were performed on DNA from 24 patients on diclofenac with hepatotoxicity, 48 subjects on diclofenac without hepatotoxicity, and healthy controls. The frequencies of the variant alleles for IL-10 and IL-4 were higher in patients (OR [odds ratio]: 2.8 for IL-10; 2.6 for IL-4; 5.3 for IL 10 + IL-4) compared with healthy controls and subjects on diclofenac without hepatotoxicity (OR: 2.8 for IL-10; 1.2 for IL-4; 5.0 for IL-10 + IL-4). In conclusion, the observed polymorphisms, resulting in low IL-10 and high IL-4 gene transcription, could favor a T helper (Th)-2 mediated antibody response to neoantigenic stimulation associated with disease susceptibility. PMID- 15122774 TI - Synbiotic modulation of gut flora: effect on minimal hepatic encephalopathy in patients with cirrhosis. AB - Minimal hepatic encephalopathy (MHE) is an important disorder that may seriously impair daily functioning and quality of life in patients with cirrhosis. Treatment with lactulose is of benefit. The possible role of synbiotics (probiotics and fermentable fiber) has not been assessed. We screened 97 consecutive cirrhotic patients without overt hepatic encephalopathy for MHE using the number connection test and measurement of brainstem auditory evoked potentials. MHE, defined by abnormality on at least one test modality, was present in 58 (60%) patients. Fifty-five of these patients with MHE were randomized to receive a synbiotic preparation (n = 20), fermentable fiber alone (n = 20), or placebo (n = 15) for 30 days. Cirrhotic patients with MHE were found to have substantial derangements in the gut microecology, with significant fecal overgrowth of potentially pathogenic Escherichia coli and Staphylococcal species. Synbiotic treatment significantly increased the fecal content of non-urease producing Lactobacillus species at the expense of these other bacterial species. Such modulation of the gut flora was associated with a significant reduction in blood ammonia levels and reversal of MHE in 50% of patients. Synbiotic treatment was also associated with a significant reduction in endotoxemia. The Child Turcotte-Pugh functional class improved in nearly 50% of cases. Treatment with fermentable fiber alone was also of benefit in a substantial proportion of patients. In conclusion, treatment with synbiotics or fermentable fiber is an alternative to lactulose for the management of MHE in patients with cirrhosis. PMID- 15122776 TI - Evolution of intrapulmonary vascular dilatations in cirrhosis. PMID- 15122778 TI - Long-term outcome of hepatitis C in children. PMID- 15122779 TI - Does noninvasive staging of fibrosis challenge liver biopsy as a gold standard in chronic hepatitis C? PMID- 15122781 TI - High sensitivity C-reactive protein values do not reliably predict the severity of histological changes in NAFLD. PMID- 15122782 TI - Prometheus and Pandora--together again. PMID- 15122786 TI - Retrospective analysis of antiretroviral HIV treatment success based on medical history or guided by the reverse hybridisation LiPA HIV genotyping system. AB - The changes in viral load and CD4(+) count at 3 and 6 months in a group of 166 HIV-infected patients was evaluated. The new therapy was chosen based on the medical history procedures for 70 patients, and in 96 patients it was guided by the partial or complete result of the line probe assay (LiPA) HIV RT and Protease resistance tests. The absolute difference from the baseline of the log viral load at 3 and 6 months was significantly different between the two groups when adjusted for baseline viral load (P < 0.0001) and stayed significant when intention-to-treat analysis was carried out (P < 0.001). The absolute difference of the CD4(+) count was not significantly different when adjusted for baseline CD4(+) (P = 0.854, 3 months; P = 0.06, 6 months). The proportion of patients with a viral load 0.5 log difference) between the two assays were detected in 39% of subtype C samples, compared to 23.2% of subtype B samples. In all cases in which a discordant result was detected, the lower results were obtained by the NucliSens assay. Discordant results between CD4 and viral load (CD4 < 200 cells/ml with a viral load <5,000 copies/ml) were observed in eight of the subtype C-infected patients when a viral load was measured by NucliSens (9.7%), compared to three patients (3.6%) when measured by the Amplicor assay. In conclusion, in patients with HIV subtype C infection, measurement of HIV RNA by the NucliSens assay resulted in a significant underestimation of the viral load as compared to the Amplicor assay. As a consequence, such an underestimation may result in sub-optimal care of patients infected with HIV subtype C. PMID- 15122789 TI - Prevalence of hepatitis A antibodies in schoolchildren in Catalonia (Spain) after the introduction of universal hepatitis A immunization. AB - The objective of this study was to investigate the prevalence of hepatitis A antibodies (anti-HAV) in schoolchildren in Catalonia and to compare it with the rates found in previous studies. Sera from a representative sample of 1,342 children aged between 6 and 15 years, recruited in 2001, were tested for anti HAV. The results were related to sociodemographic variables and vaccination history. The overall prevalence of anti-HAV was 51.4%. The prevalence was 5.5% in non-vaccinated children, similar to that found in a 1996 study, and 96.6% in vaccinated children. The prevalence of anti-HAV in non-vaccinated children increased significantly with age, reaching 11.6% in the 13-15 years age group. The prevalence of anti-HAV was higher in children born outside Catalonia than in those born in Catalonia (16.1% vs. 5.0%, P = 0.02). The expected continuation in the decline in the prevalence of anti-HAV in non-vaccinated schoolchildren, observed in Catalonia since 1986, was not found in 2001. The rate of anti-HAV in 2001 was slightly higher than in 1996, although the difference was not statistically significant (5.5 and 3.5%, respectively). This could be explained by the increased number of recent immigrant children born outside Catalonia, mainly in countries where hepatitis A is highly endemic. PMID- 15122790 TI - Hepatitis B infection of the liver in chronic hepatitis C without detectable hepatitis B virus DNA in serum. AB - Hepatitis B virus (HBV) DNA may persist in the liver in the absence of serum HBV DNA after a self-limited acute hepatitis B. This may also occur in patients with chronic hepatitis C virus (HCV) infection but its prevalence and its impact on liver histology is unknown. HBV-DNA was tested by polymerase chain reaction (PCR) and by in situ hybridisation in liver biopsies from 98 patients with chronic hepatitis C who were hepatitis B surface antigen negative and serum HBV-DNA negative by PCR. HBV-DNA resulted positive in the liver of 37/98 (37.7%) patients without serum HBV-DNA. To test whether these patients had serum HBV-DNA levels under the detection limit of the PCR assay used in this study (50 copies/ml), PCR products in which HBV-DNA was undetectable after visualization of agarose gels were analysed by dot-blot hybridisation. With this method, HBV-DNA was positive in serum of 12/37 patients with liver HBV-DNA. Thus, 25/98 (25.5%) patients have HBV-DNA detectable only in liver. This was confirmed by in situ hybridisation, the percentage of infected hepatocytes ranging from 0.1% to 12%. In patients in whom the HCV infection was shorter than 20 years, HBV infected patients had higher (P = 0.01) fibrosis score (1.64 +/- 1.21) than HBV negative cases (0.53 +/ 0.66). In conclusion, a significant proportion of patients with chronic HCV infection have HBV-DNA in the liver in the absence of viral DNA in serum. The impact of this finding on liver histology deserves further research. PMID- 15122791 TI - Reduction of double-stranded RNA-activated protein kinase in hepatocellular carcinoma associated with hepatitis B virus. AB - Chronic hepatitis B virus (HBV) infection is a major cause of hepatocellular carcinoma (HCC) in Asia. Double-stranded RNA (dsRNA)-activated protein kinase (PKR) is an interferon-induced, serine/threonine protein kinase. Recent studies have suggested that PKR is involved in the pathogenesis of HCC with hepatitis virus C infection by inhibiting viral and cellular proteins related to cell growth and proliferation. In the present study, PKR was examined in both tumor and non-tumor tissues from HCC livers infected with HBV. The expression of PKR was determined by TaqMan real-time PCR and immunohistochemical methods. The level of PKR was also analyzed in relation to pathological changes observed in HCC. The result showed that PKR was reduced in tumor tissues of HCC from HBV carriers with low serum viral load (<0.7 x 10(6) copies/ml) compared to those with higher serum viral load. However, the overall PKR level was much lower in tumor tissues than that in non-tumor tissues, irrespective of HBV carrier status or serum viral load. PKR level tended to be lower in HCC samples with alpha-fetoprotein (AFP) more than 500 ng/ml (mean: 4024.2 ng/ml) than those with AFP less than 500 ng/ml (mean: 50.6 ng/ml). There was no significant difference in the expression of PKR between tumor tissues with well differentiation and those with poor or moderate differentiation. In conclusion, the level of PKR was reduced in HCC tumor tissues, suggesting a possible role of PKR in promoting the growth of tumor. HBV may participate in altering the level of PKR, but factors other than HBV should play a more determining role in the regulation of PKR in HCC. The association between PKR and AFP levels may offer an alternative tumor marker for HCC. PMID- 15122787 TI - Molecular epidemiology of HIV in Ghana: dominance of CRF02_AG. AB - Recent studies showed the importance of CRF02_AG in West Africa, although the clinical relevance of these recombinant forms of HIV remains unknown. The present study aimed at determining the molecular diversity of HIV in Ghana and investigating the possible epidemiologic advantage of recombinant HIV-1. Plasma samples collected in 1999-2002 from two populations of HIV infected individuals (144 asymptomatic candidate blood donors and 169 AIDS patients) were studied and 249 of them were molecularly characterised in gag, pol, and env regions. Five molecular groups were identified: strains clustering with CRF02_AG in all regions (147/249 or 59%), recombinant strains clustering with CRF02_AG in one or two regions (50/249 or 20%), other subtypes, pure or recombinant, but not involving CRF02_AG (37/249 or 15%), HIV-2 (11/249 or 4.5%), and double infections (4/249 or 1.5%). There was no significant difference in the distribution of HIV-1 recombinant strains according to clinical presentation. No evidence of a significant increase in CRF02_AG prevalence between 1999 and 2002 was found. Irrespective of clinical condition, CRF02_AG is the predominant molecular form of HIV-1 in Kumasi, Ghana. PMID- 15122792 TI - Prevalence and clinical implications of occult hepatitis B viral infection in hemophilia patients in Japan. AB - The prevalence and clinical implications of occult hepatitis B virus (HBV) infection were investigated in the Japanese patients with hemophilia in whom a high prevalence of infection with transfusion-transmissible viruses has been reported. HBV DNA was detected in the sera of 22 of 43 (51.2%) patients with hemophilia who were negative for HBV surface antigen (HBs), indicating that these patients had occult HBV infection. No factor, including age, type or severity of hemophilia, presence of HBs or HBV core (HBc) antibody, or coinfection with hepatitis C virus (HCV) or human immunodeficiency virus (HIV) was associated with occult HBV infection, except for high anti-HBc titer and/or coinfection with HCV genotype 1 (1a or 1b). In general, occult HBV infection did not appear to have significant clinical implications. However, in patients in whom HBV was detected by PCR specific for the surface (S)-region, higher alanine aminotransferase levels were observed. The genotype of the occult HBV in the present study was exclusively the domestic type indigenous to Japan (genotype C), suggesting a different route of transmission for HBV in comparison to HCV and HIV in this population. PMID- 15122793 TI - Molecular and serological evaluation of surface antigen negative hepatitis B virus infection in blood donors from Venezuela. AB - Surface antigen negative hepatitis B virus (HBV) infection was evaluated in Venezuela, by molecular characterization of blood samples positive for antibodies to core antigen (anti-HBc) and negative for surface antigen (HBsAg) in blood donors (residual infections). HBV DNA was found in 11/258 samples (4.3%), and was significantly associated with high levels of anti-HBc antibodies (>25 UI/ml, P < 0.05), while no correlation was found between the presence of HBV DNA and the levels of anti-HBs. Synonymous and non-synonymous mutations were found in the HBV surface region (but not vaccine escape mutants) and in the precore/core region (precore mutants in 2/7 samples and 33-45 bp deletions near the N-terminal core region in 4/19 samples). While HBV genotype F prevails among HBsAg positive samples from blood donors in Venezuela, residual infection isolates were mainly genotypes A and D. Phylogenetic analysis of viral surface and core region revealed discrepancies in genotype designation in 6/9 samples, suggesting the presence of mixed infection or recombination. In conclusion, HBV residual infection in Venezuela does not seem to be frequently observed in HBV genotype F. This type of infection is frequently associated with variants exhibiting mutations in the surface gene that might be affecting the correct recognition by commercial tests, with precore mutants and with core internal deletions. These variants do not seem to cause severe liver disease, and on the contrary, were found circulating at low viremia. PMID- 15122794 TI - Transgenic tobacco cells producing the human monoclonal antibody to hepatitis B virus surface antigen. AB - The recombinant human monoclonal antibody (MAb) against hepatitis B virus (HBV) surface antigen (HBsAg) was expressed in tobacco suspension cultures. The parental CL4MAb was produced by the Epstein-Barr virus (EBV) transformed human cell line TAPC301-CL4. The CL4MAb cDNA was introduced into tobacco suspension cells by Agrobacterium mediated transformation. The monoclonal antibodies (MAbs), B294 and B303, which were derived from CL4 and subsequently produced in plant cells were selected for study. After purification on Protein A columns, B294 and B303 MAbs had anti-HBs relative affinity constants similar to the parental CL4MAb. B303 MAb interacted with cell surface HBsAgs and showed complement dependent cytotoxicity in a manner that was similar to anti-HBs human immunoglobulins (HBIg) that are used clinically. The results of this study point to the feasibility of producing MAbs to HBsAg in plants as an alternative to HBIg. PMID- 15122795 TI - Reliability of hepatitis C virus core antigen assay for detection of viremia in HCV genotypes 1, 2, 3, and 4 infected blood donors: a collaborative study between Japan, Egypt, and Uzbekistan. AB - Nucleic acid amplification-based methods are used for confirmation of viremia in antibody to hepatitis C virus (anti-HCV)-positive patients. However, this technology is labor intensive, time consuming, requires complex laboratory conditions, and expensive. The aim of this study was to evaluate the sensitivity and specificity of the HCV core antigen (HCVcAg) assay as an alternative approach for confirmation of viremia in HCV-infected subjects with HCV genotype 1-4. Two hundred forty-six asymptomatic HCV RNA- positive donors were enrolled in this study, consisting of 122 blood donors from Egypt (116 with genotype 4, 4 with genotype 1, and 2 with 1 + 4 genotypes), 109 from Japan (85 with genotype 1, and 24 with genotype 2), and 15 from Uzbekistan (all with genotype 3). A total of 234 (95.1%) of 246 RNA-positive specimens were detected by the HCVcAg assay; the sensitivity of HCVcAg assay consisted 93.4, 100, 100, and 94.8% for genotypes 1, 2, 3, and 4, respectively in comparison with RT-PCR assay. The specificity of the assay was confirmed in the absence of the false-positive results among 53 anti HCV-negative, but anti-Schistosoma mansoni (anti-Sm) positive donors from Egypt. A positive correlation between HCVcAg and HCV RNA concentration levels (r = 0.671, P < 0.05) was observed among specimens with HCV genotype 4. The mean HCVcAg level was significantly lower in specimens with genotype 4 (2,935 fmol/L) comparing to genotypes 1, 2, and 3 (5,034, 4,962, and 4,740 fmol/L, respectively). No specific mutation was found in the core-encoding region of the studied specimens. In conclusion, HCVcAg is shown to be specific, sensitive, and informative qualitative index for HCV viremia in asymptomatic carriers. PMID- 15122796 TI - Quantitation of cytomegalovirus DNA by the polymerase chain reaction as a predictor of disease in solid organ transplantation. AB - Cytomegalovirus (CMV) infection is an important cause of morbidity in solid organ recipients. Early markers to identify the progress of the infection and patients at high risk are required in order to apply a strategy of pre-emptive therapy. The efficacy of pre-emptive therapy relies on accurate laboratory tests to monitor CMV infection. The evaluation of CMV DNA kinetics by the polymerase chain reaction (PCR) is widely used for the management of CMV infection but markers predicting the progression of the infection and standardization of the technique are essential for the clinical interpretation of PCR results. A commercially available PCR system, the COBAS AMPLICOR Monitor (Roche Diagnostics, Brachburg, NJ), was used for the quantitation of CMV DNA in weekly blood samples (n = 504) from 47 solid organ recipients in the first 6 months after transplantation. PCR results were evaluated according to the development of clinical disease in order to find a DNA threshold and time points predicting the progression of CMV infection. Week 4 from transplantation was the earliest time point to note a significant difference between those patients who eventually developed CMV disease (n = 30) and those who remained asymptomatically infected (n = 17). At week 4, viral loads were significantly higher in patients who developed CMV disease than in asymptomatic infections (median value: 4 log(10)/10(6) leukocytes vs. 2.8, P < 0.0001). At week 4, a DNA level >/=4 log(10)/10(6) leukocytes was associated with a 45.37 odds ratio for CMV disease. Any increase >/=1 log from the first DNA detection to week 4 correlated with the clinical progression of CMV infection (odds ratio 1.74). In those patients who were treated with anti-CMV therapy, a >97% reduction of the baseline viral load was associated with a complete therapeutic success. In conclusion, CMV infection is a highly dynamic process and the quantitation of CMV DNA by PCR is a powerful marker to control successfully the infection, but a strict follow-up of the recipient and standardized PCR tests are mandatory for the best management of the infection. PMID- 15122797 TI - IL-10 levels in Dengue patients: some findings from the exceptional epidemiological conditions in Cuba. AB - The pathogenesis associated with Dengue haemorrhagic fever, has yet to be fully elucidated, with no definitive in vivo evidence. The exceptional epidemiological circumstances in Cuba allow the evaluation of different mediators in a well defined situation. In the present study, we describe the determination of levels of IL-12, IL-10 and RANTES in the sera of Cuban patients hospitalised with Dengue fever or Dengue haemorrhagic fever. The results showed that levels of serum IL-10 were higher in patients than controls, and those patients with secondary infections had consistently higher levels. All the Dengue haemorrhagic fever patients had increased levels of IL-10. In contrast, levels of IL-12 did not differ between patients and controls. Finally, RANTES serum levels detected in patients were lower than those observed in the controls. The association of increased levels of IL-10 in Dengue patients with a sequential infection suggests a possible role of this cytokine in the pathogenesis of Dengue disease. PMID- 15122799 TI - Variations in the serum concentrations of soluble Fas and soluble Fas ligand in Vietnamese patients infected with hepatitis B virus. AB - Earlier studies of both chronic hepatitis B and C virus (HBV and HCV) patients have shown a strong correlation between the soluble membrane Fas (sFas) and Fas protein expression on hepatocytes. The serum concentrations of sFas and soluble Fas ligand (sFasL) was examined in both healthy and HBV-infected Vietnamese patients to determine their relationship with the outcome of HBV infection. Patients with chronic rather than acute HBV had significantly higher amounts of sFas and sFasL, whilst the highest concentrations of both molecules were detected in those with malignant forms of HBV infection. sFas and sFasL concentrations tended to increase with a profile that paralleled the progression from asymptomatic to acute through chronic to malignant states, most markedly in the case of sFas. The sFas:sFasL ratio highlighted the relative predominance of sFas in those with acute and chronic HBV compared with asymptomatic or severe forms. In patients with hepatocellular carcinoma (HCC) a significant correlation was also observed between sFasL and alpha-feto protein (AFP) levels. The results indicate that sFas and to a lesser extent sFasL levels are to some degree associated with clinical progression in HBV infection. PMID- 15122798 TI - Limited long-term naive CD4+ T cell reconstitution in patients experiencing viral load rebounds during HAART. AB - Long-term (3.5 years) immune reconstitution in relation to viral load response was determined. Plasma HIV-1 RNA was suppressed in 40 patients (full responders) up to 42 months, and 17 patients achieved partial response. The measurements of CD4(+) and CD8(+) T lymphocyte subsets (CD45RA, CD45RACD62L, CD45RO, CD28, CD38) were carried out by flow cytometry. Full responders had a significant increase of CD4(+) and all CD4(+) T subsets both up to 6 and from 6 to 42 months, while the increase for partial responders was only up to 6 months. By 6 months, higher slopes were observed in full versus partial responders in the % of CD28 on CD4(+) and the % of CD4(+) memory subset and in both naive and memory CD4(+) subsets from 6 to 42 months. The percentage of CD8(+) and its subsets was decreased significantly in full responders both up to 6 and from 6 to 42 months (except for an increase in the CD8(+)CD45RA(+) CD62L(+) cells), while in partial responders this decrease was only up to 6 months. Lower slopes were observed in full versus partial responders from 6 to 42 months in the percentages of CD8(+), CD8(+)CD45RO(+), CD8(+)CD28(-), and CD8(+)CD38(+) T cells. In conclusion, full responders have a stronger long-term naive CD4(+) T cell subset reconstitution than partial responders. PMID- 15122800 TI - Acute retinal necrosis six years after herpes simplex encephalitis: an elusive immune deficit suggested by insufficient test sensitivity. AB - A patient presented with acute retinal necrosis of the left eye. Demonstration of herpes simplex virus (HSV) DNA in the aqueous humour confirmed the diagnosis. Negative results of HSV type-specific antibody tests based on gG antigens suggested a primary HSV infection. However, the patient had a past history of laboratory-confirmed herpes simplex encephalitis 6 years ago. Using antibody tests based on whole viral lysate antigens, he was seropositive from the onset, and immunoblot testing confirmed a lack of anti-gG reactivity. To be able to assess whether this might be related to the apparent inability of his immune system to suppress clinically symptomatic HSV infection, serial samples were tested by an HSV neutralisation test and a whole-blood flow cytometric assay to determine the frequency of HSV-specific CD4 lymphocytes. However, this did not yield evidence of obvious immunodeficiency; the patient reacted similarly to known positive controls by both assays. Although type-specific HSV serological tests based on gG are generally more specific than those based on whole viral lysate antigens, they have a somewhat lower sensitivity, as a certain percentage of HSV-infected individuals do not develop antibodies against gG, and others may suffer a secondary loss of anti-gG reactivity. Thus there is a risk of missing individual infected patients. Unless this potential problem is recognised, serious consequences might possibly result. We therefore urge virologists and clinicians to exercise great care if highly specific antibody assays based on recombinant proteins are employed. PMID- 15122801 TI - Human astrovirus, norovirus (GI, GII), and sapovirus infections in Pakistani children with diarrhea. AB - Fecal specimens from 517 infants and young children admitted to the Civil Karachi Hospital, Dow Medical College, Karachi city, Pakistan with acute gastroenteritis from 1990 to 1994 were collected and screened by RT-PCR for human astrovirus (AstV), norovirus (NV), and sapovirus (SV). The specific epidemiological data for illness caused by these viruses in Pakistan are not available. AstV, NV, and SV were detected in 58, 51, and 17 of 517 fecal specimens, and this represented 11.2, 9.9, and 3.2%, respectively. An outbreak of gastroenteritis attributable to AstV serotype 1 was identified during September and October 1990. Moreover, one specimen with a triple mixed infection between AstV (serotypes 1 and 3) and NV GII was found. NV and SV were subjected to molecular analysis by sequencing. One of the sequenced specimens positive for SV turned out to be similar to a strain tentatively called a genogroup IV. The result underscores the importance of these viruses in association with acute gastroenteritis in Karachi city, Pakistan. PMID- 15122802 TI - Human papillomavirus infection of the cervix uteri in women attending a Health Examination Center of the French social security. AB - Since human papillomavirus (HPV) is the central causal factor in cervical cancer, understanding the epidemiology of this infection constitutes an important step towards development of strategies for prevention. Six hundred and fifty seven cervical samples were tested for HPV using PCR with consensus primers (MY09/MY11), by genotyping (restriction and sequencing analyses) and by cervical cytology, from women who attended a Health Examination Center of the French social security. Women with no cervical smear as well as women with cytological abnormalities within the last 3 years were recruited. HPV DNA was detected in 7.3% of the women (5.3% for high-risk, 2.4% for low-risk, and 0.5% for unknown risk types) including 6 (0.9%) mixed infections. Fifteen different genotypes were detected, of which genotypes 16 (22.2%), 58 (13.0%), 18 (11.1%), 30 (9.2%), and 33 (9.2%) were the most prevalent. In age group 17-25 years, we found the highest frequencies for both any (22.1%) and high-risk (14.7%) HPV, and prevalences gradually decreased with age. 5.2% of low-grade squamous intraepithelial lesion, 0.3% of high-grade squamous intraepithelial lesion, and 1.2% of atypical squamous cells of undetermined significance were found. The frequencies of high risk and all HPV types were significantly higher in squamous intraepithelial lesions than in those with normal and reactive/reparative changes (P < 0.0001). The prevalence of high-risk HPV in the atypical squamous cells of undetermined significance/low grade squamous intraepithelial lesion group (28.6%) was significantly higher than in the normal and reactive/reparative changes groups (3.4%) (P < 0.0001). HPV detection was associated with younger age, single marital and non-pregnant status (P < 0.0001), premenopausal status (P = 0.0004), and contraception (P = 0.0008). Marital status (OR 4.5; 95% CI = 2.3-9.0) and tobacco consumption (OR 3.0; 95% CI = 1.6-5.7) were predictive independent factors of HPV infection. The French system of Health Examination Centers might be of interest for following women regularly, especially those with a low socioeconomic status. PMID- 15122803 TI - Influenza virological surveillance in children: the use of the QuickVue rapid diagnostic test. AB - New rapid diagnostic methods are needed to identify influenza infections to improve virological surveillance usually undertaken with conventional time consuming, complex, and even expensive laboratory methods. Another reason for using a rapid test is to avoid inappropriate therapy, particularly in children, where use of antibiotics inappropriately and high influenza-related rates of hospitalisation are described. During two winter seasons, the performance of the QuickVue Influenza test (QV) was evaluated in children under 14 presenting with influenza like illness, and compared the results with those obtained from sentinel network surveillance using standard protocols for the sample collection and the laboratory analysis by virus culture and reverse transcription-polymerase chain reaction (RT-PCR). During the first influenza season (2000/2001), 22 paediatricians collected one nose- and one throat-swab from each of the 586 children 0-6 years old recruited in the study. The QV test was carried out in the physician's office by primary care staff on the nose swab material. When compared with virus culture of the throat swab, the QV test had a sensitivity of 36.5%. In the following 2001/2002-influenza season, the performance of the QV test as a rapid laboratory screening assay was assessed. 342 children aged 0-14 years were enrolled with only one throat swab collected from each patient and sent to the laboratory where the QV, virus culture, and RT-PCR tests were performed. The results showed a better sensitivity (54.5%) of the test in comparison with virus culture and RT-PCR assays. The data indicate that rapid QV testing in the physician office setting, using these easily obtained samples, may be too insensitive to be useful for surveillance and for immediate clinical management of children presenting with influenza-like illness. Nevertheless, the QV test may be a valuable diagnostic tool if used in laboratory, as a rapid screening test. PMID- 15122804 TI - A two decade survey of respiratory adenovirus in Taiwan: the reemergence of adenovirus types 7 and 4. AB - From November 1999 to December 2001, three outbreaks of adenovirus (Ad) respiratory infection occurred in southern Taiwan. To determine the circulating serotypes and molecular epidemiology, a total of 524 virus strains were randomly selected from 1,064 strains isolated from 1981 to 2001, and were studied using restriction fragment length polymorphism (RFLP) and polymerase chain reaction (PCR)-RFLP. The major subgenus found was subgenus B (45%), followed by subgenus E (29%) and subgenus C (25%). Ad3 and Ad7 were the major types found during the 1st outbreak, which occurred from November 1999 to March 2000, while Ad4 was found mainly during the 2nd and 3rd outbreaks in October 2000 and September 2001, respectively. Both Ad7 and Ad4 were reemerged serotypes, whereas Ad3 was consistently isolated during the survey, although it declined drastically from 36 to 2% in 2001. Genotype analysis in this study showed that the only strain of Ad7 found in 1983 was Ad7a, but all randomly selected strains of Ad7 isolated during 1999-2000 were Ad7b. The clinical features of 217 patients were analyzed during the 1999-2000 outbreaks. About 79% of the total cases were less than 7 years old. The ratio of male to female was 2:1. Severe infections, such as pneumonia and acute bronchitis, accounted for nearly half of the cases (43%). These results show the reemergence and changing of serotypes, the clinical association of respiratory adenovirus infections, and the molecular epidemiology of Ad7 genotypes in Taiwan during the past two decades. PMID- 15122805 TI - Methods for the detection and characterisation of noroviruses associated with outbreaks of gastroenteritis: outbreaks occurring in the north-west of England during two norovirus seasons. AB - This article describes the methods used to investigate 407 outbreaks of acute non bacterial gastroenteritis occurring in the North-West of England between January 2000 and July 2001 and suspected to be caused by noroviruses (NV) [Mayo (2002) Arch Virol 147:1655-1663]. These included 319 outbreaks in hospitals and nursing homes and 88 other settings. Eight hundred and seventy-one faecal samples from 407 outbreaks were tested using electron microscopy (EM), an enzyme-linked immunosorbent assay (ELISA) specific for Grimsby virus (GRV) capsid antigen and/or by reverse transcriptase-polymerase chain reaction (RT-PCR) for NV, allowing the utility of each assay for routine diagnosis to be assessed. Preliminary genomic characterisation of detected strains was performed using the heteroduplex mobility assay (HMA) and DNA sequencing. The results demonstrate the continuing predominance of GII-4 GRV strain of NV as a cause of outbreaks, particularly in hospital and nursing home settings. Overall, NV were detected in 223/407 (55%) of outbreaks tested. However, a wide range of apparently diverse strains was identified, including several not previously reported. Genomic characterisation revealed clusters of linked outbreaks not recognised previously. PMID- 15122806 TI - Production of interferon gamma in respiratory syncytial virus infection of humans is not associated with interleukins 12 and 18. AB - In order to understand early events in the immune response to respiratory syncytial virus (RSV) infection, we studied the presence of various chemokines and cytokines in respiratory secretions of human infants with RSV infection. Interferon gamma (IFNgamma) was present in 30/39 (76.9%) subjects tested, but the IFNgamma-inducing cytokines interleukin (IL)12 and IL18 were detectable in 6/40 (15%) and 11/38 (28.9%) subjects, respectively. Quantities of IL12 and IL18 did not correlate with those of IFNgamma. IL18, but neither IFNgamma nor IL12 was found in significantly greater concentrations in subjects with mild, nonhypoxic forms of bronchiolitis than in those with upper respiratory illness alone or hypoxic bronchiolitis. The findings suggest that IFNgamma may be induced independently of the activities of IL12 and IL18 during RSV infection. Immune responses characterized by relatively greater release of IL18 may be associated with milder forms of bronchiolitis. PMID- 15122807 TI - Age related differences in humoral immune response to respiratory syncytial virus infection in adults. AB - The humoral immune response to Respiratory Syncytial Virus (RSV) was investigated in old and young adults. RSV was identified by culture and/or reverse transcription polymerase chain reaction (RT-PCR) in 52 elderly (mean age 74 years) and 15 young adults (mean age 33 years) with respiratory illness. Preinfection serum neutralizing and binding antibody levels were similar in the two age groups, although older persons had significantly greater serum neutralizing responses to group A RSV (log(2) 2.8 increase vs. 1.7, P = 0.001) and group B RSV (log(2) 2.8 vs. 1.3, P = 0.0008). Older persons also had significantly increased responses to the RSV F (log(2) 2.9 vs. 1.1, P < 0.0001), Ga (log(2) 2.1 vs. 1.3, P = 0.0007), and Gb proteins (log(2) 2.7 vs. 1.4, P = 0.0004). The exuberant antibody response seen in older adults may be a reflection of greater viral load and antigenic stimulation and/or a shift from Th1 to Th2 dominant immune response with aging. The relationship of immune response to disease pathogenesis in older persons deserves further study. PMID- 15122808 TI - Inhibition of respiratory syncytial virus infection with the CC chemokine RANTES (CCL5). AB - Respiratory syncytial virus (RSV) is a major cause of respiratory tract disease in infants, aged adults, and immunosuppressed patients. The only approved medicines for RSV disease are administration of prophylatic antibodies or treatment with a synthetic nucleoside. Both approaches are expensive and the latter is not without risk and of controversial benefit. The present investigation studied whether pharmaceutical or biologic compounds based upon chemokines might be useful in preventing RSV disease. Of interest was RANTES/CCL5, which inhibits infection by HIV strains that use chemokine receptor (CCR)-5 as co-receptor. Herein, we report that prior or simultaneous treatment of HEp-2 cells with recombinant human CCL5 provides dose-dependent inhibition of infection with RSV. Other recombinant chemokines (MIP-1alpha/CCL3, MIP 1beta/CCL4, MCP-2/CCL8, eotaxin/CCL11, MIP-1delta/CCL15, stromal cell derived factor (SDF)-1alpha/CXCL12) were not inhibitory. The data suggested that CCL5 might inhibit infection by blocking fusion (F) protein-epithelial cell interactions. Infections by mutant RSV strains deleted of small hydrophobic and/or attachment proteins and only expressing F protein in the envelope were inhibited by prior treatment with CCL5 or a biologically inactive N-terminally modified met-CCL5. Inhibition was also observed when virus adsorption and treatment with CCL5 were performed at 4 degrees C. Flow cytometry further revealed that epithelial cells were positive for CCR3, but not CCR1 or CCR5. Thus, novel mimetics of CCL5 may be useful prophylatic agents to prevent respiratory tract disease caused by RSV. PMID- 15122809 TI - Epidemiology of rotavirus diarrhoea in Iranian children. AB - Human rotavirus is the most important cause of severe diarrhoea in infants and young children worldwide. We describe the aetiology of viral diarrhoea and the characteristics of rotavirus infection in Shahrekord, Iran. Two hundred and fifty nine children <5 years old admitted to Hajar Hospital, 245 children with acute diarrhoea attending primary health centres in Shahrekord, and 114 children hospitalised for elective surgery were selected from October 2001 to August 2002. Stool samples were screened for enteric viruses using EM. Rotaviruses were characterised using ELISA, reverse transcription-polymerase chain reaction (RT PCR), and electropherotyping. One hundred and eighty six viruses were identified, of which 146 (78%) were rotavirus. The second most frequent virus was coronavirus, followed by calicivirus. Rotaviruses exhibited a marked seasonal variation, being most frequently isolated from November to February (50% of rotavirus recovered) and affected mostly children <2 years old. The RT-PCR successfully typed 139 of the 146 (95%) rotavirus G types and 124 (85%) P types. The most frequent P type was, P[8] in 108 (74%), P[4] in 16 (11%), and was P non typeable in 22 (15%). Among the G types, G1 was identified in 120 (82%), G2 in 19 (13%), and was G-non-typeable in 7 (5%). Our results are the first report of rotavirus genotypes affecting Iranian children. The most frequent G and P types (G1, G2, P[8], and P[4]) are similar to those reported from around the world and will be covered by existing rotavirus vaccines targeting G types G1-G4. PMID- 15122810 TI - Early inhibition of nitric oxide production increases HSV-1 intranasal infection. AB - Here, we studied the role of nitric oxide (NO) production during the first steps of the respiratory infection of BALB/c mice with herpes simplex virus type 1 (HSV 1), strain F. Nitric oxide synthase II (NOS-II) mRNA and protein were detected by reverse transcription (RT)-PCR and dot blot, respectively in samples of lungs and turbinates early post-infection (p.i.). Immunohistochemical analysis revealed pulmonar macrophages and PMN expressing NOS-II in the lungs of infected animals. Animals intranasally treated with aminoguanidine (AG), a NOS inhibitor, during the first steps of infection, showed a dose-dependent increase in pneumonitis compared to controls. Viral titres in turbinates, lungs, and brains were higher in AG treated mice. Finally, histopathology studies revealed a stronger inflammation in eyes, and lungs of these animals. Taken together, these results suggest a role of NO in controlling primary HSV intranasal infection. PMID- 15122811 TI - Soluble TREM-1 and the diagnosis of pneumonia. PMID- 15122812 TI - The Budd-Chiari syndrome. PMID- 15122813 TI - Abortion, health, and the law. PMID- 15122814 TI - Abortion, health, and the law. PMID- 15122815 TI - Paying physicians for high-quality care. PMID- 15122816 TI - Paying physicians for high-quality care. PMID- 15122817 TI - Paying physicians for high-quality care. PMID- 15122819 TI - Proceedings and abstracts from the Cardiothoracic Techniques and Technologies (CTT) 9th meeting. March 19-22, 2003, Miami, Florida, USA. PMID- 15122818 TI - Systematic response to talc pleurodesis. PMID- 15122820 TI - Review: inhaled corticosteroids slow the progression of airflow limitation in COPD. PMID- 15122821 TI - Review: long-acting beta2-agonists and inhaled corticosteroids reduce exacerbations in chronic obstructive pulmonary disease. PMID- 15122822 TI - Review: hospital-at-home care does not increase mortality or readmission rates in acute exacerbations of COPD. PMID- 15122823 TI - Review: methylxanthines are not effective for acute exacerbations of chronic obstructive pulmonary disease. PMID- 15122824 TI - Estrogen plus progestin reduced risk for fracture in postmenopausal women. PMID- 15122825 TI - Parathyroid hormone (1-84) plus alendronate was not better than monotherapy with either agent in postmenopausal osteoporosis. PMID- 15122826 TI - Parathyroid hormone (1-34) alone was better than parathyroid hormone plus alendronate in men with osteoporosis. PMID- 15122827 TI - Structured interruption of treatment hastened disease progression in multidrug resistant HIV. PMID- 15122828 TI - Buprenorphine alone or in combination with naloxone reduced the use of and craving for opiates in opiate addiction. PMID- 15122829 TI - 1-day quadruple therapy was not inferior to 7-day triple therapy for eradication of Helicobacter pylori infection in dyspepsia. PMID- 15122830 TI - D-dimer testing reduced the need for ultrasonographic imaging in outpatients with suspected deep venous thrombosis. PMID- 15122831 TI - Fondaparinux was not inferior to unfractionated heparin for symptomatic pulmonary embolism. PMID- 15122832 TI - Cannabinoids did not reduce muscle spasticity in stable multiple sclerosis. PMID- 15122833 TI - Cognitive behavioral therapy aided discontinuation of benzodiazepine in chronic insomnia. PMID- 15122834 TI - Review: antitumor necrosis factor-alpha-directed therapies are recommended for NSAID-refractory spondylorarthritis. PMID- 15122835 TI - Review: commonly used antihypertensive therapies and targeted blood pressure lowering regimens reduce cardiovascular events. PMID- 15122836 TI - Review: vitamin E, vitamin C, and possibly coenzyme Q10 are ineffective for preventing or treating cardiovascular disease. PMID- 15122837 TI - Review: spironolactone may be effective for hirsutism but data are lacking on its effectiveness for acne vulgaris in women. PMID- 15122838 TI - Review: metformin used alone or combined with clomifene may improve ovulation rates in the polycystic ovary syndrome. PMID- 15122839 TI - Review: plasma exchange or intravenous immunoglobulin reduces disability in the Guillain-Barre syndrome. PMID- 15122840 TI - An exercise and behavioral management program reduced functional dependence in Alzheimer disease. PMID- 15122842 TI - 2 fitness-related variables--exercise capacity and heart rate recovery--predicted mortality in asymptomatic women. PMID- 15122841 TI - Patients with West Nile virus meningitis or encephalitis, but not acute flaccid paralysis, had favorable outcomes. PMID- 15122843 TI - A clinical prediction model predicted 30-day and 1-year mortality in patients hospitalized for heart failure. PMID- 15122844 TI - A clinical prediction rule containing 6 variables accurately predicted hypersensitivity pneumonitis. PMID- 15122845 TI - What makes evidence-based journal clubs succeed? PMID- 15122846 TI - Acpjc.org adds enhanced searching to many other features. PMID- 15122847 TI - Campaign to revitalize academic medicine. PMID- 15122848 TI - Patients at the center: in our practice, and in our use of language. PMID- 15122850 TI - Angioplasty at an invasive-treatment center reduced mortality compared with first contact thrombolysis. PMID- 15122851 TI - Prolonged antithrombotic pretreatment increased risk for MI or death in unstable coronary syndromes. PMID- 15122852 TI - Perindopril reduced cardiac events in stable coronary artery disease. PMID- 15122853 TI - Candesartan reduced mortality and hospital admissions in chronic heart failure. PMID- 15122854 TI - Review: adjuvant corticosteroid therapy reduces death, hearing loss, and neurologic sequelae in bacterial meningitis. PMID- 15122855 TI - Noninvasive ventilation reduced duration of mechanical ventilation and ICU stay more than conventional weaning. PMID- 15122856 TI - Ethics consultation reduced hospital, ICU, and ventilation days for ICU patients who died before hospital discharge. PMID- 15122857 TI - Conventional-intensity was more effective than low-intensity warfarin therapy for preventing recurrent venous thromboembolism. PMID- 15122858 TI - High- and moderate-intensity warfarin regimens did not differ for preventing thrombosis in antiphospholipid antibody syndrome. PMID- 15122859 TI - Ximelagatran was noninferior to warfarin in preventing stroke and systemic embolism in atrial fibrillation. PMID- 15122861 TI - Review: Prophylactic acetylcysteine reduces contrast nephropathy in chronic renal insufficiency. PMID- 15122860 TI - Clarithromycin, lansoprazole, and metronidazole eradicated Helicobacter pylori infection in chronic renal insufficiency. PMID- 15122862 TI - Review: catechol O-methyl transferase inhibitors plus L-dopa and some surgical interventions improve Parkinson disease symptoms. PMID- 15122863 TI - Oral iron therapy reduced unexplained fatigue in nonanemic women with serum ferritin levels or=10-fold lower than the activity of R41Q. Acetyldithio-CoA, an analogue of the reaction product, acetyl-CoA, has been employed to test the function of arginine 41, as well as other residues (e.g., aspartate-42 and histidine-233) implicated in catalysis. Acetyldithio-CoA supports enzyme-catalyzed exchange of the methyl protons of the acetyl group with solvent; exchange is dependent on the presence of Mg(2+) and acetoacetate. In comparison with wild-type human enzyme, D42A and H233A mutant enzymes exhibit 4-fold and 10-fold decreases, respectively, in the proton exchange rate. In contrast, R41Q and R41M mutants do not catalyze any substantial enzyme-dependent proton exchange. These results suggest a role for arginine-41 in deprotonation or enolization of acetyldithio-CoA and implicate this residue in the HMG-CoA cleavage reaction chemistry that leads to acetyl-CoA product formation. Assignment of arginine-41 as an active site residue is also supported by a homology model for HMG-CoA lyase based on the structure of 4 hydroxy-2-ketovalerate aldolase. This model suggests the proximity of arginine-41 to other amino acids (aspartate-42, glutamate-72, histidine-235) implicated as active site residues based on their function as ligands to the activator cation. PMID- 15122895 TI - Conformational change of the dimeric DsbC molecule induced by GdnHCl. A study by intrinsic fluorescence. AB - Unfolding-refolding of Escherichia coli DsbC, a homodimeric molecule, induced by GdnHCl was studied by intrinsic fluorescence. Interpretation of experimental fluorescence data was done together with the analysis of protein 3D structure. It is shown that although Cys 141 is the next neighbor of the single tryptophan residue (Trp 140), the sulfur atoms of the disulfide bond Cys 141-Cys 163 are far apart from the indole ring and cannot quench its fluorescence, while the potential quenchers are Met 136 and His 170. It was revealed that though each subunit of DsbC contains eight tyrosine residues, only three tyrosine residues (Tyr 171, Tyr 38, and Tyr 52) contribute to the bulk fluorescence of the molecule. The character of intrinsic fluorescence intensity changes induced by GdnHCl (equilibrium and kinetic data) and its parametric representation, the existence of an isosbestic point of fluorescence spectra at different GdnHCl concentrations, allowed suggesting a one-step character of DsbC denaturation and its reversibility. PMID- 15122896 TI - Light-induced structural changes in a putative blue-light receptor with a novel FAD binding fold sensor of blue-light using FAD (BLUF); Slr1694 of synechocystis sp. PCC6803. AB - The sensor of blue-light using FAD (BLUF) domain is the flavin-binding fold categorized to a new class of blue-light sensing domain found in AppA from Rhodobacter sphaeroides and PAC from Euglena gracilis, but little is known concerning the mechanism of blue-light perception. An open reading frame slr1694 in a cyanobacterium Synechocystis sp. PCC6803 encodes a protein possessing the BLUF domain. Here, a full-length Slr1694 protein retaining FAD was expressed and purified and found to be present as an oligomeric form (trimer or tetramer). Using the purified Slr1694, spectroscopic properties of Slr1694 were characterized. Slr1694 was found to show the same red-shift of flavin absorption and quenching of flavin fluorescence by illumination as those of AppA. These changes reversed in the dark although the rate of dark state regeneration was much faster in Slr1694 than AppA, indicating that Slr1694 is a blue-light receptor based on BLUF with the similar photocycle to that of AppA. The dark decay in D(2)O was nearly four times slower than in H(2)O. Light-induced Fourier transform infrared (FTIR) difference spectroscopy was applied to examine the light-induced structure change of a chromophore and apo-protein with deuteration and universal (13)C and (15)N isotope labeling. The FTIR results indicate that light excitation induced distinct changes in the amide I modes of peptide backbone but relatively limited changes in flavin chromophore. Light excitation predominantly weakened the C(4)=O and C(2)=O bonding and strengthened the N1C10a and/or C4aN5 bonding, indicating formational changes of the isoalloxazine ring II and III of FAD but little formational change in the isoalloxazine ring I. The photocycle of the BLUF is unique in the sense that light excitation leads to the structural rearrangements of the protein moieties coupled with a minimum formational change of the chromophore. PMID- 15122897 TI - Severity of osteogenesis imperfecta and structure of a collagen-like peptide modeling a lethal mutation site. AB - We show that there are correlations between the severities of osteogenesis imperfecta (OI) phenotypes and changes in the residues near the mutation site. Our results show the correlations between the severity of various forms of the inherited disease OI and alteration of residues near the site of OI causing mutations. Among our many observed correlations are particularly striking ones between the presence of nearby proline residues and lethal mutations, and the presence of nearby alanines residues and nonlethal mutations. We investigated the possibility that these correlations have a structural basis using molecular dynamics simulations of collagen-like molecules designed to mimic the site of a lethal OI mutation in collagen type I. Our significant finding is that interchain hydrogen bonding is greatly affected by variations in residue type. We found that the strength of hydrogen bond networks between backbone atoms on different chains depends on the local residue sequence and is weaker in proline-rich regions of the molecule. We also found that an alanine at a site near an OI mutation causes less structural disruption than a proline, and that residue side chains also form interchain hydrogen bonds with frequencies that are dependent on residue type. For example, arginine side chains form strong hydrogen bonds with the backbone of the subsequent peptide chain, while lysine and glutamine less frequently form similar hydrogen bonds. This decrease in the observed hydrogen bond frequency correlates with a decrease in the experimentally determined thermal stability. We contrasted general structural properties of model collagen peptides with and without the mutation to examine the effect of the single-point mutation on the surrounding residues. PMID- 15122898 TI - The catalytic subunit of Escherichia coli nitrate reductase A contains a novel [4Fe-4S] cluster with a high-spin ground state. AB - We have used EPR spectroscopy, redox potentiometry, and protein crystallography to characterize the [4Fe-4S] cluster (FS0) of the Escherichia coli nitrate reductase A (NarGHI) catalytic subunit (NarG). FS0 is clearly visible in the crystal structure of NarGHI [Bertero, M. G., et al. (2003) Nat. Struct. Biol. 10, 681-687] but has novel coordination comprising one His residue and three Cys residues. At low temperatures (<15 K), reduced NarGHI exhibits a previously unobserved EPR signal comprising peaks at g = 5.023 and g = 5.556. We have assigned these features to a [4Fe-4S](+) cluster with an S = (3)/(2) ground state, with the g = 5.023 and g = 5.556 peaks corresponding to subpopulations exhibiting DeltaS = (1)/(2) and DeltaS = (3)/(2) transitions, respectively. Both peaks exhibit midpoint potentials of approximately -55 mV at pH 8.0 and are eliminated in the EPR spectrum of apomolybdo-NarGHI. The structure of apomolybdo NarGHI reveals that FS0 is still present but that there is significant conformational disorder in a segment of residues that includes one of the Cys ligands. On the basis of these observations, we have assigned the high-spin EPR features of reduced NarGHI to FS0. PMID- 15122899 TI - Perforation of the tunnel wall in carbamoyl phosphate synthetase derails the passage of ammonia between sequential active sites. AB - Carbamoyl phosphate synthetase (CPS) from Escherichia coli consists of a small subunit (approximately 42 kDa) and a large subunit (approximately 118 kDa) and catalyzes the biosynthesis of carbamoyl phosphate from MgATP, bicarbonate, and glutamine. The enzyme is able to utilize external ammonia as an alternative nitrogen source when glutamine is absent. CPS contains an internal molecular tunnel, which has been proposed to facilitate the translocation of reaction intermediates from one active site to another. Ammonia, the product from the hydrolysis of glutamine in the small subunit, is apparently transported to the next active site in the large subunit of CPS over a distance of about 45 A. The ammonia tunnel that connects these two active sites provides a direct path for the guided diffusion of ammonia and protection from protonation. Molecular damage to the ammonia tunnel was conducted in an attempt to induce leakage of ammonia directly to the protein exterior by the creation of a perforation in the tunnel wall. A hole in the tunnel wall was made by mutation of integral amino acid residues with alanine residues. The triple mutant alphaP360A/alphaH361A/betaR265A was unable to utilize glutamine for the synthesis of carbamoyl phosphate. However, the mutant enzyme retained full catalytic activity when external ammonia was used as the nitrogen source. The synchronization of the partial reactions occurring at the three active sites observed with the wild-type CPS was seriously disrupted with the mutant enzyme when glutamine was used as a nitrogen source. Overall, the catalytic constants of the mutant were consistent with the model where the channeling of ammonia has been disrupted due to the leakage from the ammonia tunnel to the protein exterior. PMID- 15122900 TI - Dissecting the catalytic mechanism of betaine-homocysteine S-methyltransferase by use of intrinsic tryptophan fluorescence and site-directed mutagenesis. AB - Betaine-homocysteine S-methyltransferase (BHMT) is a zinc-dependent enzyme that catalyzes the transfer of a methyl group from glycine betaine (Bet) to homocysteine (Hcy) to form dimethylglycine (DMG) and methionine (Met). Previous studies in other laboratories have indicated that catalysis proceeds through the formation of a ternary complex, with a transition state mimicked by the inhibitor S-(delta-carboxybutyl)-l-homocysteine (CBHcy). Using changes in intrinsic tryptophan fluorescence to determine the affinity of human BHMT for substrates, products, or CBHcy, we now demonstrate that the enzyme-substrate complex reaches its transition state through an ordered bi-bi mechanism in which Hcy is the first substrate to bind and Met is the last product released. Hcy, Met, and CBHcy bind to the enzyme to form binary complexes with K(d) values of 7.9, 6.9, and 0.28 microM, respectively. Binary complexes with Bet and DMG cannot be detected with fluorescence as a probe, but Bet and DMG bind tightly to BHMT-Hcy to form ternary complexes with K(d) values of 1.1 and 0.73 microM, respectively. Mutation of each of the seven tryptophan residues in human BHMT provides evidence that the enzyme undergoes two distinct conformational changes that are reflected in the fluorescence of the enzyme. The first is induced when Hcy binds, and the second, when Bet binds. As predicted by the crystal structure of BHMT, the amino acids Trp44 and Tyr160 are involved in binding Bet, and Glu159 in binding Hcy. Replacing these residues by site-directed mutagenesis significantly reduces the catalytic efficiency (V(max)/K(m)) of the enzyme. Replacing Tyr77 with Phe abolishes enzyme activity. PMID- 15122901 TI - Myeloproliferative disease in mice with reduced presenilin gene dosage: effect of gamma-secretase blockage. AB - Mammalian presenilins (PS) consist of two highly homologous proteins, PS1 and PS2. Because of their indispensable activity in the gamma-secretase cleavage of amyloid precursor protein to generate Abeta peptides, inhibition of PS gamma secretase activity is considered a potential therapy for Abeta blockage and Alzheimer's disease intervention. However, a variety of other substrates are also subject to PS-dependent processing, and it is thus imperative to understand the consequences of PS inactivation in vivo. Here we report a pivotal role of PS in hematopoiesis. Mice heterozygous for PS1 and homozygous for PS2 (PS1(+/)(-)PS2( )(/)(-)) developed splenomegaly with severe granulocyte infiltration. This was preceded by an overrepresentation of granulocytic cells in the bone marrow and a greatly increased multipotent granulocyte-monocyte progenitor in the spleen. In contrast, hematopoietic stem cells and T- and B-lymphocytes were not affected. Importantly, treatment of wild-type splenocytes with a gamma-secretase inhibitor directly promoted the granulocyte-macrophage colony-forming unit (GM-CFU). These results establish a critical role of PS in myelopoiesis. Our finding that this activity can be directly modulated by its gamma-secretase activity has important safety implications concerning these inhibitors. PMID- 15122902 TI - Cell-cell junctions of dermal microvascular endothelial cells contain tight and adherens junction proteins in spatial proximity. AB - Endothelial cell-cell contacts control the vascular permeability, thereby regulating the flow of solutes, macromolecules, and leukocytes between blood vessels and interstitial space. Because of specific needs, the endothelial permeability differs significantly between the tight blood-brain barrier endothelium and the more permeable endothelial lining of the non-brain microvasculature. Most likely, such differences are due to a differing architecture of the respective interendothelial cell contacts. However, while the molecules and junctional complexes of macrovascular endothelial cells and the blood-brain barrier endothelium are fairly well characterized, much less is known about the organization of intercellular contacts of microvascular endothelium. Toward this end, we developed a combined cross-linking and immunoprecipitation protocol which enabled us to map nearest neighbor interactions of junctional proteins in the human dermal microvascular endothelial cell line HMEC-1. We show that proteins typically located in tight or adherens junctions of epithelial cells are in the proximity in HMEC-1 cells. This contrasts with the separation of the different types of junctions observed in polarized epithelial cells and "tight" endothelial layers of the blood-brain barrier and argues for a need of the specific junctional contacts in microvascular endothelium possibly required to support an efficient transendothelial migration of leukocytes. PMID- 15122903 TI - Influence of electric charge variation at residues 209 and 159 on the interaction of eIF4E with the mRNA 5' terminus. AB - Eukaryotic translation initiation factor 4E (eIF4E) is essential for efficient protein synthesis in cap-dependent translation. The protein specifically binds the cap structure at the mRNA 5' terminus and facilitates the assembly of the mRNA with other initiation factors and the 40S ribosomal subunit. Phosphorylation of eIF4E is implicated in the regulation of the initiation step of translation. However, the molecular mechanism of this regulation still remains unclear. To address this problem, we have determined the binding affinities of eIF4E specifically mutated at position 209 or 159 for a series of novel mono- and dinucleotide cap analogues by a fluorometric time-synchronized titration method. A 1.5-3-fold reduction in the affinity of cap for the S209E mutant and a 1-2-fold increase in the affinity of cap for the S209K mutant, depending on the negative charge of phosphate chains, indicate that phosphorylation at Ser209 creates electrostatic repulsion between the protein and the negatively charged cap structure. The inhibition of the ability to bind cap analogues by the K159A mutant and its phosphorylated counterpart shows significant participation of Lys159 in the binding of the capped mRNA. Both structural modifications, phosphorylation and the replacement of lysine with alanine, result in an increase in the negative Gibbs free energy of association that is proportional to the length of the cap phosphate chain and additive, i.e., equal to the sum of the individual destabilizing changes of DeltaG degrees. The possible implication of these results for the mechanism of control of eIF4E by phosphorylation, especially for the "clamping model", is discussed. PMID- 15122904 TI - Evidence that both kinetic and thermodynamic factors govern DNA toroid dimensions: effects of magnesium(II) on DNA condensation by hexammine cobalt(III). AB - Millimolar concentrations of divalent cations are shown to affect the size of toroids formed when DNA is condensed by multivalent cations. The origins of this effect were explored by varying the order in which MgCl(2) was added to a series of DNA condensation reactions with hexammine cobalt chloride. The interplay between Mg(II), temperature, and absolute cation concentration on DNA condensation was also investigated. These studies reveal that DNA condensation is extremely sensitive to whether Mg(II) is associated with DNA prior to condensation or Mg(II) is added concurrently with hexammine cobalt(III) at the time of condensation. It was also found that, in the presence of Mg(II), temperature and dilution can have opposite effects on the degree of DNA condensation. A systematic comparison of DNA condensates observed in this study clearly illustrates that, under our low-salt conditions, toroid size is determined by the kinetics of toroid nucleation and growth. However, when Mg(II) is present during condensation, toroid size can also be limited by a thermodynamic parameter (e.g., undercharging). The path dependence of DNA condensation presented here illustrates that regardless of which particular factors limit toroid growth, toroids formed under the various conditions of this study are largely nonequilibrium structures. PMID- 15122905 TI - Sequence-dependent thermodynamic parameters for locked nucleic acid (LNA)-DNA duplex formation. AB - The design of modified nucleic acid probes, primers, and therapeutics is improved by considering their thermodynamics. Locked nucleic acid (LNA) is one of the most useful modified backbones, with incorporation of a single LNA providing a substantial increase in duplex stability. In this work, the hybridization DeltaH(o), DeltaS(o), and melting temperature (T(M)) were measured from absorbance melting curves for 100 duplex oligonucleotides with single internal LNA nucleotides on one strand, and the results provided DeltaDeltaH(o), DeltaDeltaS(o), DeltaDelta, and DeltaT(M) relative to reference DNA oligonucleotides. LNA pyrimidines contribute more stability than purines, especially A(L), but there is substantial context dependence for each LNA base. Both the 5' and 3' neighbors must be considered in predicting the effect of an LNA incorporation, with purine neighbors providing more stability. Enthalpy entropy compensation in DeltaDeltaH(o) and DeltaDeltaS(o) is observed across the set of sequences, suggesting that LNA can stabilize the duplex by either preorganization or improved stacking, but not both simultaneously. Singular value decomposition analysis provides predictive sequence-dependent rules for hybridization of singly LNA-substituted DNA oligonucleotides to their all-DNA complements. The results are provided as sets of DeltaDeltaH(o), DeltaDeltaS(o), and DeltaDelta parameters for all 32 of the possible nearest neighbors for LNA+DNA:DNA hybridization (5' MX(L) and 5' X(L)N, where M, N, and X = A, C, G, or T and X(L) represents LNA). The parameters are applicable within the standard thermodynamic prediction algorithms. They provide T(M) estimates accurate to within 2 degrees C for LNA-containing oligonucleotides, which is significantly better accuracy than previously available. PMID- 15122906 TI - Quantitative analyses of bifunctional molecules. AB - Small molecules can be discovered or engineered to bind tightly to biologically relevant proteins, and these molecules have proven to be powerful tools for both basic research and therapeutic applications. In many cases, detailed biophysical analyses of the intermolecular binding events are essential for improving the activity of the small molecules. These interactions can often be characterized as straightforward bimolecular binding events, and a variety of experimental and analytical techniques have been developed and refined to facilitate these analyses. Several investigators have recently synthesized heterodimeric molecules that are designed to bind simultaneously with two different proteins to form ternary complexes. These heterodimeric molecules often display compelling biological activity; however, they are difficult to characterize. The bimolecular interaction between one protein and the heterodimeric ligand (primary dissociation constant) can be determined by a number of methods. However, the interaction between that protein-ligand complex and the second protein (secondary dissociation constant) is more difficult to measure due to the noncovalent nature of the original protein-ligand complex. Consequently, these heterodimeric compounds are often characterized in terms of their activity, which is an experimentally dependent metric. We have developed a general quantitative mathematical model that can be used to measure both the primary (protein + ligand) and secondary (protein-ligand + protein) dissociation constants for heterodimeric small molecules. These values are largely independent of the experimental technique used and furthermore provide a direct measure of the thermodynamic stability of the ternary complexes that are formed. Fluorescence polarization and this model were used to characterize the heterodimeric molecule, SLFpYEEI, which binds to both FKBP12 and the Fyn SH2 domain, demonstrating that the model is useful for both predictive as well as ex post facto analytical applications. PMID- 15122907 TI - Aeromonas proteolytica aminopeptidase: an investigation of the mode of action using a quantum mechanical/molecular mechanical approach. AB - The aminopeptidase of Aeromonas proteolytica (AAP) belongs to the group of metallo-hydrolases that require two divalent cations for full activity. Such binuclear metal centers are found in several aminopeptidases, raising the question whether a common mechanism, at least partly, is likely. We have used a quantum mechanical/molecular mechanical (QM/MM) approach to investigate the reaction mechanism of AAP. Among several possibilities, one reaction path was found to be clearly the most favorable. Beside the chemical transformation steps, effects of the enzyme environment and the influence of the solvent on the catalytic reaction were included in the study. The results are in good agreement with experimental studies and correspond to a high degree to our previous QM/MM calculations on the reaction mechanism of the related binuclear bovine lens leucine aminopeptidase (blLAP), which, although related to the AAP, has different Zn(2+)-coordination spheres and a different catalytic residue. The mechanisms of the two enzymes as suggested in the literature differ on the mode of coordination of the nucleophile and the identity of the general base. However, the results of this and our previous work on blLAP allow us to identify a common mechanism for the two enzymes. This mechanism is probably quite general for binuclear zinc enzymes. PMID- 15122908 TI - Domain structures and roles in bacteriophage HK97 capsid assembly and maturation. AB - Head assembly in the double-stranded DNA coliphage HK97 involves initially the formation of the precursor shell Prohead I from approximately 420 copies of a 384 residue subunit. This is followed by proteolytic removal of residues 2-103 to create Prohead II, and then reorganization and expansion of the shell lattice and covalent cross-linking of subunits make Head II. Here, we report and structurally interpret solution Raman spectra of Prohead I, Prohead II, and Head II particles. The Raman signatures of Prohead I and Prohead II indicate a common alpha/beta fold for residues 104-385, and a strongly conserved tertiary structure. The Raman difference spectrum between Prohead I and Prohead II demonstrates that the N terminal residues 2-103 (Delta-domain) form a predominantly alpha-helical fold devoid of beta-strand. The conformation of the Delta-domain in Prohead I thus resembles that of the previously characterized scaffolding proteins of Salmonellaphage P22 and Bacillus phage phi29 and suggests an analogous architectural role in mediating the assembly of a properly dimensioned precursor shell. The Prohead II --> Head II transition is accompanied by significant reordering of both the secondary and tertiary structures of 104-385, wherein a large increase occurs in the percentage of beta-strand (from 38 to 45%), and a marginal increase is observed in the percentage of alpha-helix (from 27 to 31%). Both are at the expense of unordered chain segments. Residue environments affected by HK97 shell maturation include the unique cysteine (Cys 362) and numerous tyrosines and tryptophans. The tertiary structural reorganization is reminiscent of that observed for the procapsid --> capsid transformation of P22. The Raman signatures of aqueous and crystalline Head II reveal no significant differences between the crystal and solution structures. PMID- 15122909 TI - Metal ion affinities of the zinc finger domains of the metal responsive element binding transcription factor-1 (MTF1). AB - Metal response element (MRE) binding transcription factor-1 (MTF1) is a six Cys(2)His(2) zinc finger-containing transcription factor required for basal and zinc-induced transcription of metallothionein genes. The cobalt(II) and zinc(II) affinities of a protein fragment comprising the six zinc finger domains have been examined to reveal apparent dissociation constants (for the six domains collectively) of 0.5 +/- 0.2 microM for cobalt(II) and 31 +/- 14 pM for zinc(II). Two approaches have been used to determine the metal ion affinities of the individual domains. First, the six domains have been examined as single domain peptides revealing dissociation constants ranging from 0.3 to 1.7 microM for cobalt(II). The domains fall into two sets with peptides corresponding to domains 2, 3, and 4 showing relatively high affinity (K(d)(Co(II)) 0.3-0.5 microM) and peptides corresponding to domains 1, 5, and 6 showing lower affinity (K(d)(Co(II)) 1.6-1.7 microM). Second, we examined the affinity of each domain in the context of the six zinc finger domain protein by individually mutating one metal-binding His residue to Cys to allow independent monitoring of the cobalt(II) occupancy of each site. The affinity of each domain was higher in this context than as a single domain peptide with affinities (corrected for the effect of the mutation) ranging from 0.02 to 0.5 microM. The increase in affinity for the individual domains ranged from factors of 1.1 to 20. The order of affinities (from higher to lowest) was observed to be 4 > 2 approximately 5 > 6 approximately 3 approximately 1. These results reveal that none of the Cys(2)His(2) zinc finger domains of MTF1 have dramatically low metal ion affinities, certainly none low enough to respond to changes in free zinc ion concentrations in the micromolar range. Nonetheless, the metal ion affinities of some domains do differ by a factor of 25 with domains at both the amino- and carboxyl-termini showing lower intrinsic affinities for metal ions than the central domains. PMID- 15122910 TI - Signal transduction mechanism of a peptide mimetic of interferon-gamma. AB - The C-terminus of interferon-gamma (IFNgamma) contains a nuclear localization sequence (NLS) required for the activation and nuclear translocation of the transcription factor STAT1alpha and induction of IFNgamma-activated genes. On the basis of this and other studies, we developed a peptide mimetic of IFNgamma that possesses the IFNgamma functions of antiviral activity and upregulation of MHC class II molecules. The mimetic also shares with IFNgamma the ability to induce the activation and nuclear translocation of STAT1alpha and the IFNgamma receptor (IFNGR)-1 subunit. The mimetic, IFNgamma(95-132), is a peptide that consists of the C-terminal residues 95-132 of murine IFNgamma and contains a required alpha helical domain and the NLS of IFNgamma. In this study, we determined the mechanism of the intracellular action of the mimetic at the level of signal transduction. We show that the mimetic mediates the nuclear transport of IFNGR-1 through its interaction with IFNGR-1 cytoplasmic region 253-287 via both the helical region and the NLS of IFNgamma(95-132). Alanine substitutions of the NLS of the mimetic showed that the NLS was required for nuclear translocation and that the nuclear transport properties of the mimetic correlated with its ability to bind IFNGR-1. These data also show that the NLS of IFNgamma(95-132) can interact simultaneously with IFNGR-1 and the nuclear import machinery. We found that in in vitro nuclear transport assays tyrosine-phosphorylated STAT1alpha failed to undergo nuclear translocation in the presence of nuclear import factors, but was transported to nucleus in the presence of IFNgamma(95-132) and JAK2-phosphorylated IFNGR-1, to which STAT1alpha binds, as a complex of IFNgamma(95-132)/IFNGR-1/STAT1alpha. Thus, the mimetic, which possesses IFNgamma function, is directly involved as a chaperone in the nuclear transport of STAT1alpha and shares this mechanism of action with that previously described for IFNgamma. The mimetic, like IFNgamma, is able to upregulate the tumor suppressor p21WAF1/CIP1, a direct target of STAT1alpha, and this ability requires the NLS of the mimetic. However, unlike IFNgamma, the mimetic is unable to downregulate c myc and hence does not inhibit the cycling of cells. This suggests that IFNgamma has additional functions that are not tied directly to the nuclear translocation of STAT1alpha. PMID- 15122911 TI - Conversion of the 2,2,6,6-tetramethylpiperidine moiety to a 2,2 dimethylpyrrolidine by cytochrome P450: evidence for a mechanism involving nitroxide radicals and heme iron. AB - Earlier we described a novel cytochrome P450 (CYP) catalyzed metabolism of the 2,2,6,6-tetramethylpiperidine (2,2,6,6-TMPi) moiety in human liver microsomes to a ring-contracted 2,2-dimethylpyrrolidine (2,2-DMPy) [Yin, W., et al. (2003) Drug Metab. Dispos. 31, 215-223]. In the current report, evidence is provided for the involvement of 2,2,6,6-TMPi hydroxylamines and their one-electron oxidation products, the nitroxide radicals, as intermediates in this pathway. Nitroxide radicals could be converted to their corresponding 2,2-DMPy metabolites by "inactivated CYP3A4", as well as by a number of other heme proteins and hemin, suggesting that this is a heme-catalyzed process. The conversion of nitroxide radicals to the 2,2-DMPy products by CYP3A4 or hemin was accompanied by the generation of acetone in incubations, providing evidence that the three-carbon unit from 2,2,6,6-TMPi was lost as acetone. With one model 2,2,6,6-TMPi nitroxide radical, evidence for an alternate pathway, which resulted in the formation of an intermediate that incorporated two oxygen atoms from water of the incubation medium before collapsing to the 2,2-DMPy product, was also obtained. To account for both pathways, a mechanism involving interaction of the nitroxide radicals with heme iron (Fe(III)), followed by a homolytic scission of the N-O bond and transfer of the nitroxide oxygen to heme iron to form a perferryl-oxygen complex, is proposed. The nitrogen-centered 2,2,6,6-TMPi radical thus formed then precipitates the contraction of the piperidine ring via C2-C3 bond cleavage, and the resulting product further oxidizes to an exocyclic iminium ion (by the perferryl-oxygen complex); the latter may undergo capture by water from the incubation medium and eliminate the three-carbon unit via N-dealkylation. It remains to be determined whether this novel interaction of nitroxide radicals with heme iron has any relevance in regard to the known biological properties of these stable radical species. PMID- 15122912 TI - Single amino acids in the lumenal loop domain influence the stability of the major light-harvesting chlorophyll a/b complex. AB - The major light-harvesting complex of photosystem II (LHCIIb) is one of the most abundant integral membrane proteins. It greatly enhances the efficiency of photosynthesis in green plants by binding a large number of accessory pigments that absorb light energy and conduct it toward the photosynthetic reaction centers. Most of these pigments are associated with the three transmembrane and one amphiphilic alpha helices of the protein. Less is known about the significance of the loop domains connecting the alpha helices for pigment binding. Therefore, we randomly exchanged single amino acids in the lumenal loop domain of the bacterially expressed apoprotein Lhcb1 and then reconstituted the mutant protein with pigments in vitro. The resulting collection of mutated recombinant LHCIIb versions was screened by using a 96-well-format plate-based procedure described previously [Heinemann, B., and Paulsen, H. (1999) Biochemistry 38, 14088-14093], enabling us to test several thousand mutants for their ability to form stable pigment-protein complexes in vitro. At least one third of the positions in the loop domain turned out to be sensitive targets; i.e., their exchange abolished formation of LHCIIb in vitro. This confirms our earlier notion that the LHCIIb loop domains contribute more specifically to complex formation and/or stabilization than by merely connecting the alpha helices. Among the target sites, glycines and hydrophilic amino acids are more prominently represented than hydrophobic ones. Specifically, the exchange of any of the three acidic amino acids in the lumenal loop abolishes reconstitution of stable pigment-protein complexes, suggesting that ionic interactions with other protein domains are important for correct protein folding or complex stabilization. One hydrophobic amino acid, tryptophan in position 97, has been hit repeatedly in independent mutation experiments. From the LHCIIb structure and previous mutational analyses, we propose a stabilizing interaction between this amino acid and F195 near the C-proximal end of the third transmembrane helix. PMID- 15122913 TI - Expression, purification, and characterization of Bacillus subtilis cytochromes P450 CYP102A2 and CYP102A3: flavocytochrome homologues of P450 BM3 from Bacillus megaterium. AB - The cyp102A2 and cyp102A3 genes encoding the two Bacillus subtilis homologues (CYP102A2 and CYP102A3) of flavocytochrome P450 BM3 (CYP102A1) from Bacillus megaterium have been cloned, expressed in Escherichia coli, purified, and characterized spectroscopically and enzymologically. Both enzymes contain heme, flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN) cofactors and bind a variety of fatty acid molecules, as demonstrated by conversion of the low spin resting form of the heme iron to the high-spin form induced by substrate binding. CYP102A2 and CYP102A3 catalyze the fatty acid-dependent oxidation of reduced nicotinamide adenine dinucleotide phosphate (NADPH) and reduction of artificial electron acceptors at high rates. Binding of carbon monoxide to the reduced forms of both enzymes results in the shift of the heme Soret band to 450 nm, confirming the P450 nature of the enzymes. Reverse-phase high-performance liquid chromatography (HPLC) of products from the reaction of the enzymes with myristic acid demonstrates that both catalyze the subterminal hydroxylation of this substrate, though with different regioselectivity and catalytic rate. Both P450s 102A2 and 102A3 show kinetic and binding preferences for long-chain unsaturated and branched-chain fatty acids over saturated fatty acids, indicating that the former two molecule types may be the true substrates. P450s 102A2 and 102A3 exhibit differing substrate selectivity profiles from each other and from P450 BM3, indicating that they may fulfill subtly different cellular roles. Titration curves for binding and turnover kinetics of several fatty acid substrates with P450s 102A2 and 102A3 are better described by sigmoidal (rather than hyperbolic) functions, suggesting binding of more than one molecule of substrate to the P450s, or possibly cooperativity in substrate binding. Comparison of the amino acid sequences of the three flavocytochromes shows that several important amino acids in P450 BM3 are not conserved in the B. subtilis homologues, pointing to differences in the binding modes for the substrates that may explain the unusual sigmoidal kinetic and titration properties. PMID- 15122914 TI - Analysis of the early stage of the folding process of reduced lysozyme using all lysozyme variants containing a pair of cysteines. AB - Twenty-eight hen lysozyme variants that contained a pair of cysteines were constructed to examine the formation of the individual native and nonnative disulfide bonds. We analyzed the extent of the formation of a disulfide bond in each lysozyme variant using a redox buffer (pH 8) containing 1.0 mM reduced and 0.1 mM oxidized glutathione in the absence or presence of 6 M guanidine hydrochloride. In the presence of 6 M guanidine hydrochloride, the extent of the formation of the disulfide bond in each lysozyme variant was proportional to the distance between cysteine residues, indicating that reduced hen lysozyme under a highly denaturing condition adopted a randomly coiled structure. In aqueous solution, the formations of all disulfide bonds occurred much more easily than under a denatured condition. This finding indicated that reduced lysozyme had a somewhat compact structure. Moreover, the scattering data for the extents of the formation of the disulfide bonds among all lysozyme variants were observed. These results suggested that the nonrandom folding occurred in the early stage of the folding of reduced lysozyme, which should provide new insight into the early stage events in the folding process of reduced lysozyme. PMID- 15122915 TI - Further insights into quinone cofactor biogenesis: probing the role of mauG in methylamine dehydrogenase tryptophan tryptophylquinone formation. AB - Paracoccus denitrificans methylamine dehydrogenase (MADH) is an enzyme containing a quinone cofactor tryptophan tryptophylquinone (TTQ) derived from two tryptophan residues (betaTrp(57) and betaTrp(108)) within the polypeptide chain. During cofactor formation, the two tryptophan residues become covalently linked, and two carbonyl oxygens are added to the indole ring of betaTrp(57). Expression of active MADH from P. denitrificans requires four other genes in addition to those that encode the polypeptides of the MADH alpha(2)beta(2) heterotetramer. One of these, mauG, has been shown to be involved in TTQ biogenesis. It contains two covalently attached c-type hemes but exhibits unusual properties compared to c type cytochromes and diheme cytochrome c peroxidases, to which it has some sequence similarity. To test the role that MauG may play in TTQ maturation, the predicted proximal histidine to each heme (His(35) and His(205)) has each been mutated to valine, and wild-type MADH was expressed in the background of these two mauG mutants. The resultant MADH has been characterized by mass spectrometry and electrophoretic and kinetic analyses. The majority species is a TTQ biogenesis intermediate containing a monohydroxylated betaTrp(57), suggesting that this is the natural substrate for MauG. Previous work has shown that MADH mutated at the betaTrp(108) position (the non-oxygenated TTQ partner) is predominantly also this intermediate, and work on these mutants is extended and compared to the MADH expressed in the background of the histidine to valine mauG mutations. In this study, it is unequivocally demonstrated that MauG is required to initiate the formation of the TTQ cross-link, the conversion of a single hydroxyl located on betaTrp(57) to a carbonyl, and the incorporation of the second oxygen into the TTQ ring to complete TTQ biogenesis. The properties of MauG, which are atypical of c-type cytochromes, are discussed in the context of these final stages of TTQ biogenesis. PMID- 15122916 TI - Characterization of bis(levuglandinyl) urea derivatives as products of the reaction between prostaglandin H2 and arginine. AB - Levuglandins are gamma-keto aldehydes formed by rearrangement of prostaglandin (PG) H(2) in aqueous solution. Levuglandins are highly reactive with primary amines. We had previously characterized adducts formed after reaction of levuglandin E(2) (LGE(2)) or PGH(2) with lysine. In this study, we assessed whether reaction of PGH(2) with arginine yielded covalent adducts. Using N(alpha) acetylarginine and both PGH(2) and synthetic LGE(2), we discovered a novel series of levuglandinyl adducts derived from reaction of two levuglandin moieties with the guanidino group of arginine. Subsequent spontaneous hydrolysis of the adducted amino acid yields bis(levuglandinyl) urea and the corresponding ornithine residue. Using liquid chromatography tandem mass spectrometry, we characterized the molecular structure of these novel adducts and demonstrated their formation after coincubation of PGH(2) with synthetic peptides and proteins. The soluble characteristic of these molecules provides a potential strategy for development of biological markers of lipid modification of proteins following cyclooxygenase activity or lipid peroxidation. PMID- 15122917 TI - Cloning and characterization of mouse nucleoside triphosphate diphosphohydrolase 8. AB - A novel mammalian plasma membrane bound nucleoside triphosphate diphosphohydrolase (NTPDase), named NTPDase8, has been cloned and characterized. Analysis of cDNA reveals an open reading frame of 1491 base pairs encoding a protein of 497 amino acid residues with an estimated molecular mass of 54650 Da and a predicted isoelectric point of 5.94. In a mouse, the genomic sequence is located on chromosome 2A3 and is comprised of 10 exons. The deduced amino acid sequence reveals eight putative N-glycosylation sites, two transmembrane domains, five apyrase-conserved regions, and 20-50% amino acid identity with other mammalian NTPDases. mRNA expression was detected in liver, jejunum, and kidney. Both intact cells and crude cell lysates from COS-7 cells expressing NTPDase8 hydrolyzed P2 receptor agonists, namely, ATP, ADP, UTP, and UDP, but did not hydrolyze AMP. There was an absolute requirement for divalent cations for the catalytic activity (Ca(2+) > Mg(2+)) with an optimal pH between 5.5 and 8.0 for ATP and 6.4 for ADP hydrolysis. Kinetic parameters derived from analysis of crude cell lysates showed that the enzyme had lower apparent K(m) values for adenine nucleotides and for triphosphonucleosides (K(m,app) of 13 microM for ATP, 41 microM for ADP, 47 microM for UTP, and 171 microM for UDP). Hydrolysis of triphosphonucleosides resulted in a transient accumulation of the corresponding diphosphonucleoside, as expected from the apparent K(m) values. Enzymatic properties of NTPDase8 differ from those of other NTPDases suggesting an alternative way to modulate nucleotide levels and consequently P2 receptor activation. PMID- 15122918 TI - Poly(ethylene glycol)-induced fusion and destabilization of human plasma high density lipoproteins. AB - High-density lipoproteins (HDL) are macromolecular complexes of specific proteins and lipids that mediate the removal of cholesterol from peripheral tissues. Chemical unfolding revealed that HDL fusion and rupture are the two main kinetic steps in HDL denaturation. Here we test the hypothesis that lipid fusogens such as poly(ethylene glycol) (PEG) may promote lipoprotein fusion and rupture and thereby destabilize HDL. We analyze thermal disruption of spherical HDL in 0-15% PEG-8000 by calorimetric, spectroscopic, electron microscopic, and light scattering techniques. We demonstrate that the two irreversible high-temperature endothermic HDL transitions involve particle enlargement and show a heating rate dependence characteristic of kinetically controlled reactions with high activation energy. The first calorimetric transition reflects HDL fusion and dissociation of lipid-poor apolipoprotein A-1 (apoA-1), and the second transition reflects HDL rupture and release of the apolar lipid core. Neither transition involves substantial protein unfolding; thus, the transition heat originates from lipid and/or protein dissociation and repacking. At room temperature, PEG-8000 induces HDL fusion that is distinct from the heat-, denaturant-, or enzyme induced fusion since it leads to formation of larger particles and does not involve apoA-1 dissociation. Increasing the PEG concentration in solution from 0 to 15% leads to low-temperature shifts by approximately -18 degrees C in the two calorimetric HDL transitions without altering their nature. Thus, consistent with our hypothesis, PEG-8000 induces fusion and reduces the thermal stability of HDL. Our results suggest that PEG is useful for the analysis of the molecular events involved in metabolic HDL remodeling and fusion. PMID- 15122919 TI - Role of the 9-methyl group of retinal in cone visual pigments. AB - In rhodopsin, the 9-methyl group of retinal has previously been identified as being critical in linking the ligand isomerization with the subsequent protein conformational changes that result in the activation of its G protein, transducin. Here, we report studies on the role of this methyl group in the salamander rod and cone pigments. Pigments were generated by combining proteins expressed in COS cells with 11-cis 9-demethyl retinal, where the 9-methyl group on the polyene chain has been deleted. The absorption spectra of all pigments were blue-shifted. The red cone and blue cone/green rod pigments were unstable to hydroxylamine; whereas, the rhodopsin and UV cone pigments were stable. The lack of the 9-methyl group of the chromophore did not affect the ability of the red cone and blue cone/green rod pigments to activate transducin. On the other hand, with the rhodopsin and UV cone pigments, activation was diminished. Interestingly, the red cone pigment containing the retinal analogue remained active longer than the native pigment. Thus, the 9-methyl group of retinal is not important in the activation pathway of the red cone and blue cone/green rod pigments. However, for the red cone pigment, the 9-methyl group of retinal appears to be critical in the deactivation pathway. PMID- 15122920 TI - Differential cytotoxicity of human wild type and mutant alpha-synuclein in human neuroblastoma SH-SY5Y cells in the presence of dopamine. AB - Parkinson's disease (PD) involves loss of dopaminergic neurons in the substantia nigra and is characterized by intracellular inclusions, Lewy bodies, consisting primarily of aggregated alpha-synuclein. Two substitution mutations (A53T and A30P) in alpha-synuclein gene have been identified in familial early-onset PD. To understand the biological changes that incur upon alpha-synuclein-induced cytotoxicity in the presence of dopamine, the current studies were undertaken. Human SH-SY5Y neuroblastoma cells coexpressing the human dopamine transporter [hDAT], and either wild type (wt) or mutant alpha-synucleins, were treated with 50 microM dopamine (DA). In cells expressing wt or A30P alpha-synuclein, DA accelerated production of reactive oxygen species and cell death as compared to cells expressing A53T or hDAT alone. The increased sensitivity of such cells to DA was investigated by measuring changes in cellular ionic gradient, by atomic absorption spectrometry, and cell metabolism, by high-resolution nuclear magnetic resonance spectroscopy. Both wt and A30P alpha-synuclein caused rapid decrease in levels of intracellular potassium, followed by mitochondrial damage and cytochrome c leakage, with decreased cellular metabolism as compared to cells expressing A53T or hDAT alone. Collapse of ionic gradient was significantly faster in A30P (t(1/2) = 3.5 h) than in wt (t(1/2) = 6.5 h) cells, and these changes in ionic gradient preceded cytochrome c leakage and depletion of metabolic energy. Neither wt nor mutant alpha-synuclein resulted in significant changes in ionic gradient or cellular metabolism in the absence of intracellular DA. These findings suggest a specific sequence of events triggered by dopamine and differentially exacerbated by alpha-synuclein and the A30P mutant. PMID- 15122921 TI - Decavanadate binding to a high affinity site near the myosin catalytic centre inhibits F-actin-stimulated myosin ATPase activity. AB - Decameric vanadate (V(10)) inhibits the actin-stimulated myosin ATPase activity, noncompetitively with actin or with ATP upon interaction with a high-affinity binding site (K(i) = 0.27 +/- 0.05 microM) in myosin subfragment-1 (S1). The binding of V(10) to S1 can be monitored from titration with V(10) of the fluorescence of S1 labeled at Cys-707 and Cys-697 with N-iodo-acetyl-N'-(5-sulfo 1-naphthyl)ethylenediamine (IAEDANS) or 5-(iodoacetamido) fluorescein, which showed the presence of only one V(10) binding site per monomer with a dissociation constant of 0.16-0.7 microM, indicating that S1 labeling with these dyes produced only a small distortion of the V(10) binding site. The large quenching of AEDANS-labeled S1 fluorescence produced by V(10) indicated that the V(10) binding site is close to Cys-697 and 707. Fluorescence studies demonstrated the following: (i) the binding of V(10) to S1 is not competitive either with actin or with ADP.V(1) or ADP.AlF(4); (ii) the affinity of V(10) for the complex S1/ADP.V(1) and S1/ADP.AlF(4) is 2- and 3-fold lower than for S1; and (iii) it is competitive with the S1 "back door" ligand P(1)P(5)-diadenosine pentaphosphate. A local conformational change in S1 upon binding of V(10) is supported by (i) a decrease of the efficiency of fluorescence energy transfer between eosin-labeled F-actin and fluorescein-labeled S1, and (ii) slower reassociation between S1 and F-actin after ATP hydrolysis. The results are consistent with binding of V(10) to the Walker A motif of ABC ATPases, which in S1 corresponds to conserved regions of the P-loop which form part of the phosphate tube. PMID- 15122922 TI - Paramagnetism-based refinement strategy for the solution structure of human alpha parvalbumin. AB - In the frame of a research aimed at the detailed structural characterization of human calcium-binding proteins of the EF-hand family, the solution structure of human alpha-parvalbumin has been solved by NMR and refined with the help of substitution of the Ca(2+) ion in the EF site with the paramagnetic Dy(3+) ion. A simple (1)H-(15)N HSQC spectrum allowed the NH assignments based on the properties of Dy(3+). This allowed us to exploit pseudocontact shifts and residual dipolar couplings for solution structure refinement. The backbone and heavy atom RMSD are 0.55 +/- 0.08 and 1.02 +/- 0.08 A, respectively, and decrease to 0.39 +/- 0.05 and 0.90 +/- 0.06 A upon refinement with paramagnetism-based restraints. The RMSD for the metal itself in the EF site in the refined structure is 0.26 +/- 0.12 A. Backbone NH R(1), R(2), and NOE measured at two temperatures show the protein to be relatively rigid. The NH orientations are well determined by the paramagnetism-based restraints. This allows us to detect small but significant local structural differences with the orthologue protein from rat, whose X-ray structure is available at 2.0 A resolution. All differences are related to local changes in the amino acidic composition. PMID- 15122924 TI - Acute stressors and cortisol responses: a theoretical integration and synthesis of laboratory research. AB - This meta-analysis reviews 208 laboratory studies of acute psychological stressors and tests a theoretical model delineating conditions capable of eliciting cortisol responses. Psychological stressors increased cortisol levels; however, effects varied widely across tasks. Consistent with the theoretical model, motivated performance tasks elicited cortisol responses if they were uncontrollable or characterized by social-evaluative threat (task performance could be negatively judged by others), when methodological factors and other stressor characteristics were controlled for. Tasks containing both uncontrollable and social-evaluative elements were associated with the largest cortisol and adrenocorticotropin hormone changes and the longest times to recovery. These findings are consistent with the animal literature on the physiological effects of uncontrollable social threat and contradict the belief that cortisol is responsive to all types of stressors. PMID- 15122925 TI - The costly pursuit of self-esteem. AB - Researchers have recently questioned the benefits associated with having high self-esteem. The authors propose that the importance of self-esteem lies more in how people strive for it rather than whether it is high or low. They argue that in domains in which their self-worth is invested, people adopt the goal to validate their abilities and qualities, and hence their self-worth. When people have self-validation goals, they react to threats in these domains in ways that undermine learning; relatedness; autonomy and self-regulation; and over time, mental and physical health. The short-term emotional benefits of pursuing self esteem are often outweighed by long-term costs. Previous research on self-esteem is reinterpreted in terms of self-esteem striving. Cultural roots of the pursuit of self-esteem are considered. Finally, the alternatives to pursuing self-esteem, and ways of avoiding its costs, are discussed. PMID- 15122926 TI - The healthy pursuit of self-esteem: comment on and alternative to the Crocker and Park (2004) formulation. AB - J. Crocker and L. E. Park (2004) proposed both (a) that the importance of self esteem lies more in how people strive for it rather than whether it is high or low and (b) that the benefits of pursuing self-esteem are of a limited, short term nature and are outweighed by the significant costs that entail for functioning in multiple areas. The authors review research that raises questions about the validity of each of these assumptions. They conclude that findings are more consistent with a view in which (a) high self-esteem, or at least the avoidance of low self-esteem, is an important factor contributing to overall health and well-being and (b) the pursuit of self-esteem, when directed toward adaptive ends, can be instrumental in promoting long-term outcomes that are of value to both individuals and society. They also describe an integrative theoretical framework that encompasses both of these possibilities. PMID- 15122927 TI - The benefits of a "sidelong" approach to self-esteem need satisfaction: comment on Crocker and Park (2004). AB - J. Crocker and L. E. Park (2004) have achieved an admirable integration of the self-esteem literature with their claim that self-esteem is better conceived of as a dynamic human striving, rather than as a passive state or personality characteristic. However, the costs of self-esteem striving may be overstated- these costs may arise only in certain constrained cases. Also, although Crocker and Park suggested that self-esteem is not a true psychological need, there is evidence that humans in all cultures need to feel a positive sense of self-worth (K. M. Sheldon, in press). Problems may arise only when people strive too directly for this feeling, rather than deriving it as a natural concomitant of non-self-focused goals. A "sidelong" approach to self-esteem need satisfaction is advocated in this commentary. PMID- 15122928 TI - Can we really do without self-esteem? Comment on Crocker and Park (2004). AB - Because self-esteem plays a central role in managing anxiety and provides a superordinate goal that organizes much human behavior, the authors conclude that it would be extremely difficult and probably impossible for people to stop pursuing self-esteem. Although they agree that investment in others' welfare may circumvent some of the negative consequences of self-esteem pursuit, the authors argue that the self-esteem implications of other-oriented behavior will inevitably be a powerful motivating force. Finally, the authors propose that self esteem derived from self-determined standards of value may reduce defensiveness, closed-mindedness, and indifference to others, as well as promote personal growth, but they acknowledge the daunting barriers to the attainment of such self determined bases of self-worth. PMID- 15122930 TI - Why do people need self-esteem? A theoretical and empirical review. AB - Terror management theory (TMT; J. Greenberg, T. Pyszczynski, & S. Solomon, 1986) posits that people are motivated to pursue positive self-evaluations because self esteem provides a buffer against the omnipresent potential for anxiety engendered by the uniquely human awareness of mortality. Empirical evidence relevant to the theory is reviewed showing that high levels of self-esteem reduce anxiety and anxiety-related defensive behavior, reminders of one's mortality increase self esteem striving and defense of self-esteem against threats in a variety of domains, high levels of self-esteem eliminate the effect of reminders of mortality on both self-esteem striving and the accessibility of death-related thoughts, and convincing people of the existence of an afterlife eliminates the effect of mortality salience on self-esteem striving. TMT is compared with other explanations for why people need self-esteem, and a critique of the most prominent of these, sociometer theory, is provided. PMID- 15122931 TI - Do people need self-esteem? Comment on Pyszczynski et al. (2004). AB - In spite of impressive empirical evidence consistent with aspects of terror management theory (TMT) reviewed by T. Pyszczynski, J. Greenberg, S. Solomon, J. Arndt, and J. Schimel (2004), several fundamental assumptions of the theory remain untested or lack support. Specifically, Pyszczynski et al. (2004) have not demonstrated that (a) people need self-esteem, (b) pursuing self-esteem is an effective means for reducing anxiety, (c) pursuing self-esteem helps people achieve their important goals, (d) having or pursuing self-esteem is the only way to deal with anxiety to achieve important goals, or (e) death is the real issue driving the pursuit of self-esteem. The authors suggest there is a different paradigm for thinking about death, one in which awareness of one's mortality serves as a precious reminder of the limited time one has to accomplish one's most important goals. All of these questions can be addressed with empirical research. PMID- 15122932 TI - Avoiding death or engaging life as accounts of meaning and culture: comment on Pyszczynski et al. (2004). AB - Terror management theory emphasizes that self-esteem consists of a sense of meaning and significance, which serves mainly to defend against death awareness. The current authors counter that people's search for meaning and significance cannot be wholly reduced to defensive processes because it also reflects intrinsic developmental processes. Sociometer theory similarly offers a mainly defensive account of self-esteem, and its exclusive focus on belongingness versus exclusion ill equips it to deal with the multiple needs underlying self-esteem. The current authors suggest that self-esteem resulting from defenses against anxiety (whether about death or exclusion) is akin to contingent self-esteem, whereas true self-esteem is based in ongoing satisfaction of needs for competence, autonomy, and relatedness. PMID- 15122933 TI - The function of self-esteem in terror management theory and sociometer theory: comment on Pyszczynski et al. (2004). AB - By applying different standards of evidence to sociometer theory than to terror management theory (TMT), T. Pyszczynski, J. Greenberg, S. Solomon, J. Arndt, and J. Schimel's (2004) review offers an imbalanced appraisal of the theories' merits. Many of Pyszczynski et al.'s (2004) criticisms of sociometer theory apply equally to TMT. and others are based on misconstruals of the theory or misunderstandings regarding how people respond when rejected. Furthermore, much of their review is only indirectly relevant to TMT's position on the function of self-esteem, and the review fails to acknowledge logical and empirical challenges to TMT. A more balanced review suggests that each theory trumps the other in certain respects, both have difficulty explaining all of the evidence regarding self-esteem, and the propositions of each theory can be roughly translated into the concepts of the other. For these reasons, declaring a theoretical winner at this time is premature. PMID- 15122935 TI - Conceptualizing and measuring personality vulnerability to depression: comment on Coyne and Whiffen (1995). AB - J. C. Coyne and V. E. Whiffen (1995) reviewed research on personality vulnerability to depression, focusing on S. J. Blatt's (1974, 1990) concepts of dependency and self-criticism and A. T. Beck's (1983) concepts of sociotropy and autonomy. The authors discuss 6 issues raised in that review: (a) the typological or dimensional nature of vulnerability, (b) the theoretical implications of "mixed" vulnerability, (c) the relations of vulnerability to Neuroticism. (d) the potential confounding of vulnerability with concurrent depression, (e) the potential confounding of vulnerability with social context, and (f) the differentiation of dependency from relatedness. The authors conclude that Blatt's and Beck's concepts are continuous, nearly orthogonal dimensions that can be identified and measured independently from Neuroticism, depression, and social context. PMID- 15122938 TI - Memory accessibility, mood regulation, and dysphoria: difficulties in repairing sad mood with happy memories? AB - Recent studies have suggested that mood-incongruency effects are due to mood regulatory processes, in which people retrieve positive memories to repair negative moods. In Study 1, the authors investigated whether dysphoria influences the accessibility of autobiographical memories following a positive or a negative mood induction combined with subsequent rumination or distraction. The results showed a mood-repair effect for nondysphoric but not for dysphoric participants following rumination. In Study 2, participants were asked to either distract themselves or to recall positive autobiographical memories after a negative mood induction. Whereas nondysphoric participants' mood improved under both conditions, dysphoric participants' mood improved only after distraction. These results suggest that dysphoria is associated with a reduced ability to use mood incongruent recall to repair sad moods. PMID- 15122939 TI - All symptoms are not created equal: the prominent role of hyperarousal in the natural course of posttraumatic psychological distress. AB - This 3-wave longitudinal study examined the natural course of posttraumatic stress disorder symptoms using data collected from young adult survivors of community violence. Three key findings emerged. 1. Mean levels of distress for each symptom cluster decreased over time, with reexperiencing decreasing most rapidly. 2. Cross-lagged panel analysis revealed that hyperarousal strongly influences, but is not generally influenced by, other symptoms clusters. 3. Trajectory analysis demonstrated that respondents for whom hyperarousal was the most pronounced baseline symptom showed lower overall symptom improvement relative to trauma exposed counterparts for whom hyperarousal was a less prominent early symptom. Implications for theory, research, and clinical practice are discussed. PMID- 15122940 TI - Parent-adolescent disagreement regarding psychopathology in adolescents from the general population as a risk factor for adverse outcome. AB - This study investigated whether parent-adolescent disagreement regarding adolescents' behavioral and emotional problems predicted adverse outcome. A Dutch sample of 15- to 18-year-olds was prospectively followed across a 4-year interval. The Child Behavior Checklist (CBCL: T. M. Achenbach, 1997) and Youth Self-Report (YSR; T. M. Achenbach, 1991b) were administered at initial assessment, and the following signs of poor outcome were assessed 4 years later: police/judicial contacts, expulsion from school/job, suicidal ideation, unwanted pregnancy, suicide attempts, deliberate self-harm, referral to mental health services, report of having a behavioral or emotional problem, and feeling the need for professional help without actually receiving help. Twenty CBCL syndrome scores, 23 YSR syndrome scores, and 16 discrepancy scores were significant predictors of poor outcome. It was concluded that to determine the prognosis of psychopathology in adolescents, discrepancies between informants may be important. PMID- 15122941 TI - The Smell Identification Test as a measure of olfactory identification ability in schizophrenia and healthy populations: a Rasch psychometric study. AB - This study examines University of Pennsylvania Smell Identification Test (UPSIT; R. L. Doty, 1995) performance in 133 controls and 54 chronic, medicated outpatients with schizophrenia (SZ) using item-response theory modeling. Results show that UPSIT items contribute to 1 factor, cover a range of 8 standard errors of measurement, and articulate 3 ability levels. Although it is not difficult enough to discriminate among persons of above-average ability, the test has diagnostic utility in detecting moderate impairment. Independent of item difficulty, 13 items differentiate patients from controls. When 45 patients and 45 controls were matched on gender and age, patient accuracy remained significantly reduced. The findings support the test's utility and demonstrate how traditional data analysis is insensitive to complexities in test performance. PMID- 15122942 TI - The longitudinal relationship of personality traits and disorders. AB - Personality disorders are presumed to be stable because of underlying stable and maladaptive personality traits, but while previous research has demonstrated a link between personality traits and personality disorders cross-sectionally, personality disorders and personality traits have not been linked longitudinally. This study explores the extent to which relevant personality traits are stable in individuals diagnosed with 4 personality disorders (schizotypal, borderline, avoidant, and obsessive-compulsive personality disorders) and examines the assumption that these personality disorders are stable by virtue of stable personality traits. This assumption was tested via the estimation of a series of latent longitudinal models that evaluated whether changes in relevant personality traits lead to subsequent changes in personality disorders. In addition to offering large consistency estimates for personality traits and personality disorders, the results demonstrate significant cross-lagged relationships between trait change and later disorder change for 3 of the 4 personality disorders studied. PMID- 15122943 TI - A psychophysical measure of attention deficit in children with attention deficit/hyperactivity disorder. AB - The authors explored the temporal mechanism of attention deficit in children with attention-deficit/hyperactivity disorder (ADHD). In rapid serial visual presentation tasks in which two targets (T-sub-1 and T-sub-2) were presented in close temporal proximity among distractors, participants tried to identify T-sub 1 and detect T-sub-2 in one (dual-task) experiment and only to detect T-sub-2 in a second control (single-task) experiment. The sensitivity of T-sub-2 detection was analyzed using signal detection theory. The attentional blink--the impairment in T-sub-2 detection following the identification of T-sub-1--was increased in magnitude and protracted in the patients. Moreover, some ADHD children appeared to have a blink largely normal in magnitude but temporally displaced toward a later time. The authors hypothesize that a slower closing of the attention gate may mediate this specific attention impairment in ADHD children. PMID- 15122944 TI - Risk for separation anxiety disorder among girls: paternal absence, socioeconomic disadvantage, and genetic vulnerability. AB - The authors examined genetic and environmental influences, including the contributions of 2 measured aspects of the shared environment of twins (paternal absence, socioeconomic disadvantage) on the development of mother-reported separation anxiety disorder (SAD) history in a sample of 1,887 female twin pairs. Four different symptom categories of SAD were considered. Results revealed that all 4 SAD symptom categories were significantly heritable, whereas the contribution of shared environmental influences to the variation in risk was significant for only 2 of the 4 SAD categories. Paternal absence was found to have an important influence in vulnerability for SAD, whereas the effect of socioeconomic disadvantage was less robust. Evidence for race differences in the etiology of SAD was not found. PMID- 15122945 TI - A cross-cultural investigation of cognitions and depressive symptoms in adolescents. AB - Adolescents (N=2,272) from Hong Kong and the United States provided information regarding their depressive symptoms, cognitions (self-efficacy, negative cognitive errors, and hopelessness), and stressful events between 2 surveys 6 months apart. Depressive symptoms and hopelessness were higher, and self-efficacy and negative cognitive errors were lower in Hong Kong than in the United States. Cognitions were associated with concurrent depressive symptoms and predicted depressive symptoms 6 months later in both cultures. The "reverse" model was also supported with more variance predicted by depressive symptoms to later cognitions than from cognitions to depressive symptoms. There was some support for the hypothesis that self-efficacy is less salient in collective compared with individualistic cultures. These findings extend cognitive theories of depression to a non-Western culture. PMID- 15122946 TI - Clinically abusive relationships in an unselected birth cohort: men's and women's participation and developmental antecedents. AB - In an unselected birth cohort (N=980, age 24-26 years), individuals in abusive relationships causing injury and/or official intervention (9% prevalence) were compared with participants reporting physical abuse without clinical consequences and with control participants who reported no abuse, on current characteristics and prospective developmental risks. In nonclinically abusive relationships, perpetrators were primarily women. In clinically abusive relationships, men and women used physical abuse, although more women needed medical treatment for injury. Women in clinically abusive relationships had childhood family adversity, adolescent conduct problems, and aggressive personality; men had disinhibitory psychopathology since childhood and extensive personality deviance. These findings counter the hibitory assumption that if clinical abuse was ascertained in epidemiological samples, it would be primarily man-to-woman, explained by patriarchy rather than psychopathology. PMID- 15122947 TI - ALDH2 status and conduct disorder mediate the relationship between ethnicity and alcohol dependence in Chinese, Korean, and White American college students. AB - This study examined aldehyde dehydrogense (ALDH2) gene status, alcohol dehydrogense (ADH2) gene status, conduct disorder, and alcohol dependence in Chinese, Korean, and White American college students. Chinese had a lower rate of alcohol dependence (5%) than Koreans (13%) and Whites (17%). Koreans had a higher rate of conduct disorder (15%) than Whites (9%) and Chinese (6%). The relationship of ethnicity to alcohol dependence was mediated by ALDH2 status and conduct disorder, although Chinese ethnicity remained significant. ADH2 status was not related to alcohol dependence with ALDH2 included, and no interactions were significant. Results suggest that different rates of risk (e.g., conduct disorder) and protective (e.g., ALDH2 status) factors partially account for ethnic differences in rates of alcohol dependence. PMID- 15122948 TI - Borderline personality disorder features predict negative outcomes 2 years later. AB - In a sample of 351 young adults, the authors assessed whether borderline personality disorder (BPD) features prospectively predicted negative outcomes (poorer academic achievement and social maladjustment) over the subsequent 2 years, over and above gender and both Axis I and Axis II psychopathology. Borderline traits were significantly related to these outcomes, with impulsivity and affective instability the most highly associated. The present findings suggest that the impulsivity and affective instability associated with BPD leads to impairment in relating well with others, in meeting social role obligations, and in academic or occupational achievement. Therefore, these may be especially important features to target in interventions for BPD. PMID- 15122949 TI - Attention, heart rate, and startle response during exposure to trauma-relevant pictures: a comparison of recent trauma victims and patients with posttraumatic stress disorder. AB - Victims of a recent trauma were compared with posttraumatic stress disorder (PTSD) patients and healthy controls to assess whether a specific anxiety response and an attentional bias were evident initially or only in chronic PTSD. Heart rate (HR) and startle response were measured, and a dot-probe task was carried out using trauma-relevant pictures. Severely affected recent trauma victims and chronic PTSD patients showed HR acceleration to trauma-related material, which was the only significant group difference. A bias away from trauma-related material was related to severity of intrusions in recent trauma victims, and the bias toward trauma-related material increased with amplitude of the HR response in PTSD patients. A specific anxiety reaction is present initially in severely affected trauma victims. PMID- 15122950 TI - Neuropsychological executive functioning in children at elevated risk for alcoholism: findings in early adolescence. AB - One component of individual risk for alcoholism may involve cognitive vulnerabilities prodromal to alcoholism onset. This prospective study of 198 boys followed between 3 and 14 years of age evaluated neurocognitive functioning across three groups who varied in familial risk for future alcoholism. Measures of intelligence, reward-response, and a battery of neuropsychological executive and cognitive inhibitory measures were used. Executive functioning weaknesses were greater in families with alcoholism but no antisocial comorbidity. IQ and reward-response weaknesses were associated with familial antisocial alcoholism. Executive function effects were clearest for response inhibition, response speed, and symbol-digit modalities. Results suggest that executive deficits are not part of the highest risk, antisocial pathway to alcoholism but that some executive function weaknesses may contribute to a secondary risk pathway. PMID- 15122951 TI - Suppression of attentional bias in PTSD. AB - Sixty combat veterans with posttraumatic stress disorder performed an emotional Stroop task under 1 of 4 contextual conditions designed to test theoretical explanations for an attentional bias suppression effect. Results revealed that when the emotional Stroop task was performed under conditions involving a future threat of either watching a combat video or giving a speech, attentional bias was inhibited. There was limited support for the prediction that the suppression effect was strongest when stressor content matched word content on the Stroop. In contrast to participants in the threat conditions, veterans who believed that they would receive additional compensation for speeded color naming or who believed that they would have no other experimental demands were slower when color naming combat-threat words. Potential theoretical explanations of the findings are discussed. PMID- 15122952 TI - PTSD arousal and depression symptoms associated with increased right-sided parietal EEG asymmetry. AB - Researchers have proposed that depression and particular types of anxiety are associated with unique patterns of regional brain activation. The authors examined the relationship among posttraumatic stress disorder (PTSD), anxiety, and depressive symptoms and frontal, temporal, and parietal EEG alpha asymmetry in female Vietnam War nurse veterans. The results indicate that PTSD arousal symptoms are associated with increased right-sided parietal activation. However, the combination of arousal, depression, and their interaction explain more than twice the variance in parietal asymmetry compared with arousal alone. The results support the contention that the association between anxiety and right-sided posterior activation is specific to the anxious arousal subtype. These findings underscore the importance of isolating, both theoretically and statistically, emotional subcomponents in studies of regional brain activation. PMID- 15122953 TI - The influence of instructions and nicotine dose on the subjective and reinforcing effects of smoking. AB - Subjective and reinforcing effects of smoking a cigarette were examined within a 2 x 2 modified balanced-placebo design, which manipulated instructions about nicotine content (i.e., told regular nicotine vs. told low nicotine) and actual nicotine dose (given a regular nicotine brand vs. a denicotinized brand). Most ratings of the nicotine content and reward value of cigarettes were higher for those told regular nicotine versus told low nicotine, and for those given regular nicotine versus given low nicotine. Nicotine and instructions did not affect craving, withdrawal, or smoke-reinforced responding, but instructions affected the number of puffs earned for those given low nicotine (i.e., placebo effect). Thus, verbal information (instructions) can influence some responses to smoking consistent with the presence of placebo and antiplacebo effects. PMID- 15122954 TI - Using the false memory paradigm to test two key elements of alcohol expectancy theory. AB - Two key aspects of alcohol expectancy theory--(a) that memories about alcohol effects are stored as relatively cohesive templates of information and (b) that these templates are automatically activated in alcohol-related contexts--were tested using the Deese-Roediger- McDermott false memory paradigm. Alcohol expectancy adjectives were studied, and false memory for expectancy target words was tested in neutral and alcohol contexts. Results indicated that in the alcohol context heavier drinkers showed more false memory for alcohol expectancy words than they did in a neutral context. Differences were not found for lighter drinkers. These results were consistent with alcohol expectancy theory, which was then compared with various forms of association theory in explaining these results and larger issues in the addiction field. PMID- 15122955 TI - Differential effects of bremazocine on oral phencyclidine (PCP) self administration in male and female rhesus monkeys. AB - Sex differences exist in many phases of drug abuse, but few studies have focused on sex differences in drug abuse treatment. In this study, the effects of bremazocine, a kappa-opioid receptor agonist, were compared in age-matched male and female rhesus monkeys self-administering orally delivered phencyclidine (PCP). Bremazocine (0.00032. 0.001, and 0.0025 mg/kg, intramuscular) was administered for 5 consecutive days. 15 min prior to daily 3-hr sessions when PCP (0.25 mg/ml) and water were available under concurrent fixed-ratio schedules. Bremazocine dose-dependently decreased PCP-maintained responding and consumption (mg/kg) in males and females, and these measures were suppressed at a lower bremazocine dose in females than in males. The percentage reduction in PCP maintained responding and intake (mg/kg) was significantly greater in females than it was in males at the low and middle doses of bremazocine, suggesting that females may be more responsive to kappa agonist treatment than males. PMID- 15122956 TI - Alcohol's dissociation of implicit and explicit memory processes: implications of a parallel distributed processing model of semantic priming. AB - Alcohol's dissociation of implicit (unintentional) and explicit (intentional) memory processes in social drinkers was examined. It was hypothesized that an alcohol challenge would lower the percentage of words recalled and result in more retroactive interference in explicit recall tasks but would not lengthen reaction time in an implicit semantic priming task involving highly semantically similar words. Men and women completed all memory tasks in each of 2 counterbalanced sessions (alcohol challenge vs. no-alcohol) separated by 1 week. Alcohol significantly degraded processing in both explicit memory tasks, yet implicit semantic priming remained intact. A parallel distributed processing model that simulates semantic memory is presented. When this system is strongly activated, it does not appear to be altered during moderate alcohol intoxication. PMID- 15122957 TI - Participants receiving dehydroepiandrosterone during treatment for cocaine dependence show high rates of cocaine use in a placebo-controlled pilot study. AB - Twenty-three cocaine-dependent participants were randomly assigned to receive either dehydroepiandrosterone (DHEA; n = 11; 100 mg/day) or placebo (n = 12) in the context of 12 weeks of thrice weekly cognitive-behavioral group counseling. Outcomes were retention, urine drug screening, cocaine craving, adverse experiences, and medication compliance. DHEA-treated participants averaged 45.8 (SD = 28.8) days in treatment, compared with 70.7 (SD = 20.6) days for placebo, r(21) = -2.4, p =.03, and provided 26.8% (SD = 29.3) of urine samples free of cocaine metabolite compared with 70.6% (SD = 39.9) for the placebo condition, r(21) = -3.0, p =.01. No differences were detected between conditions for cocaine craving or adverse experiences. High levels of medication compliance were documented. Results argue against using high doses of DHEA as a pharmacotherapy for cocaine dependence. PMID- 15122958 TI - Limitations in the assessment of DSM-IV cannabis tolerance as an indicator of dependence in adolescents. AB - The usefulness of the Diagnostic and Statistical Manual's (4th ed.; DSM-IV; American Psychiatric Association, 1994) tolerance criterion as an indicator of dependence has been debated. The authors of this study evaluated the performance of DSM's cannabis tolerance criterion, operationally defined as a percentage increase in quantity needed to get high, in distinguishing adolescents with and without cannabis dependence. Two samples of adolescent cannabis users (ages 12 19) provided data (ns = 417 and 380). Tolerance, defined as a percentage increase (median increase = 300% and 175%, respectively, in the samples), had only moderate overall sensitivity and specificity in distinguishing those with and without cannabis dependence. Results suggest limitations of the DSM-IV and change based operational definition of tolerance in adolescents. PMID- 15122959 TI - A tale of two cities: financing two voucher programs for substance abusers through community donations. AB - Voucher-based reinforcement therapy (VBRT) is an effective drug abuse treatment, but the cost of VBRT rewards has limited its dissemination. Obtaining VBRT incentives through donations may be one way to overcome this barrier. Two direct mail campaigns solicited donations for use in VBRT for pregnant, postpartum, and parenting drug users in Toronto, Ontario, Canada, and in Los Angeles, California. In Toronto, 19% of those contacted over 2 months donated 8,000 dollars (4,000 dollars/month) of goods and services. In Los Angeles, nearly 26% of those contacted over 34 months donated 161,000 dollars (4,472dollars/month) of goods and services. Maintaining voucher programs by soliciting donations is feasible and sustainable. The methods in this article can serve as a guide for successful donation solicitation campaigns. Donations offer an alternative for obtaining VBRT rewards for substance abuse treatment and may increase its dissemination. PMID- 15122960 TI - Correlates of attachment at age 3: construct validity of the preschool attachment classification system. AB - This study examined correlates of attachment at age 3 to further validate preschool separation-reunion measures. Three-year-olds (N = 150) and their mothers participated in a separation-reunion protocol, the Preschool Attachment Classification System (PACS: J. Cassidy & R. S. Marvin with the MacArthur Working Group on Attachment, 1992), and a mother-child interaction session during a laboratory visit. Mothers also completed psychosocial measures and, along with teachers, evaluated child behavior problems. The secure and disorganized groups received, respectively, the highest and lowest interaction scores. Disorganized children showed a higher level of teacher-reported externalizing and internalizing problems than did secure children. Mothers of insecure children reported higher child externalizing (all insecure groups) and internalizing (avoidant group) scores, more personal distress related to emotional bonding (disorganized group), childrearing control (ambivalent group), and child hyperactivity (avoidant group). Results strongly support the validity of the PACS as a measure of attachment in 3-year-olds. PMID- 15122962 TI - Chinese children's effortful control and dispositional anger/frustration: relations to parenting styles and children's social functioning. AB - Relations among authoritative and authoritarian parenting styles, children's effortful control and dispositional anger/frustration, and children's social functioning were examined for 425 first and second graders (7-10 years old) in Beijing, China. Parents reported on parenting styles; parents and teachers rated children's effortful control, anger/frustration, externalizing problems, and socially appropriate behaviors: and peers rated aggression and leadership/sociability. High effortful control and low dispositional anger/frustration uniquely predicted Chinese children's high social functioning, and the relation of anger/frustration to social functioning was moderated by effortful control. Authoritarian parenting was associated with children's low effortful control and high dispositional anger/frustration, which (especially effortful control) mediated the negative relation between authoritarian parenting and children's social functioning. Effortful control weakly mediated the positive relation of authoritative parenting to social functioning. PMID- 15122961 TI - Genetic and environmental influences on mothering of adolescents: a comparison of two samples. AB - This study examined 2 samples of adolescents and mothers using a child-based design (Nonshared Environment in Adolescent Development [NEAD] project, N = 395 families) and a parent-based design (Twin Moms [TM] project, N = 236 twin family pairs) to compare genetic and environmental influences on mothering. For both samples, the same measures of positivity, negativity, control, and monitoring were used. The use of matched child-based and parent-based samples enabled passive and nonpassive genotype-environment (GE) correlations to be approximated, providing information about process. Passive GE correlations were suggested for mother's positivity and monitoring. For mother's negativity and control, primarily nonpassive GE correlations were suggested. In several cases, both types of GE correlation were indicated. Finally, observer ratings of negativity and monitoring were influenced only by environmental factors. PMID- 15122963 TI - The relations between persistent poverty and contextual risk and children's behavior in elementary school. AB - Does persistent adversity over time have effects on children's behavior beyond the effects of intermittent or concurrent adversity? This study examined the relations between school behavior in 5th grade (mean age = 11 years 0 months) and indexes representing persistent poverty and contextual risk. The indexes described 2-year intervals of family adversity. The results showed effects for persistent risk over 2 consecutive intervals for several factors, but only for recent intervals (3rd and 5th grades), and the factors differed for externalizing behavior, internalizing behavior, and academic competence. The 3rd interval of risk added little to the outcomes, and most factors did not show persistence effects. The results highlight the need for caution in expecting and interpreting effects for persistent adversity. PMID- 15122964 TI - Overt and relational aggression and perceived popularity: developmental differences in concurrent and prospective relations. AB - Relations of overt and relational aggression with perceived popularity among children and early adolescents were examined in 2 studies (Ns = 607 and 1,049). Among older youths, positive concurrent relations found between overt aggression and perceived popularity became nonsignificant when relational aggression was controlled, whereas positive associations found between relational aggression and perceived popularity held when overt aggression was controlled. Aggression and perceived popularity were not positively related for the younger participants. The 2nd study also examined the temporal ordering of these relations over 6 months. For older girls, positive relations between relational aggression and perceived popularity were bidirectional. For older boys, relational aggression did not predict increased perceived popularity, but perceived popularity predicted increased relational aggression. Implications for intervention are discussed. PMID- 15122965 TI - Young children know that trying is not pretending: a test of the "behaving-as-if" construal of children's early concept of pretense. AB - In 3 studies, young children were tested for their understanding of pretend actions. In Studies 1 and 2, pairs of superficially similar behaviors were presented to 26- and 36-month-old children in an imitation game. In one case the behavior was marked as trying (signs of effort), and in the other case as pretending (signs of playfulness). Three-year-olds, and to some degree 2-year olds, performed the real action themselves (or tried to really perform it) after the trying model, whereas after the pretense model, they only pretended. Study 3 ruled out a simple mimicking explanation by showing that children not only imitated differentially but responded differentially with appropriate productive pretending to pretense models and with appropriate productive tool use to trying models. The findings of the 3 studies demonstrate that by 2 to 3 years of age, children have a concept of pretense as a specific type of intentional activity. PMID- 15122966 TI - Families created through surrogacy arrangements: parent-child relationships in the 1st year of life. AB - Findings are presented of a study of families created through surrogacy arrangements. Forty-two surrogacy families were compared with 51 egg-donation families and 80 natural-conception families on standardized interview and questionnaire measures of the psychological well-being of the parents, the quality of parent-child relationships, and infant temperament. The differences that were identified between the surrogacy families and the other family types indicated greater psychological well-being and adaptation to parenthood by mothers and fathers of children born through surrogacy arrangements than by the natural-conception parents. PMID- 15122967 TI - Children's mental health in times of economic recession: replication and extension of the family economic stress model in Finland. AB - This study evaluated the applicability of the family economic stress model (FESM) in understanding the influences of economic hardship on child mental health during a nationwide economic recession in Finland. The information was gathered from 527 triads of 12-year-olds and their mothers and fathers from a population sample. The structural equation models showed that the FESM fit the data well, indicating its generalizability in Finnish society. The results confirmed that a reduction in disposable family income constitutes a risk for child mental health through increased economic pressure and negative changes in parental mental health, marital interaction, and parenting quality. Controlling the children's prerecession mental health substantiated that economic hardship can lead to deterioration in children's mental health. Alternative models based on fully recursive analyses revealed reciprocal influences between parents and their children over time: Children's prerecession mental health problems predicted compromised parenting, which in turn contributed to children's internalizing and externalizing symptoms during the recession. PMID- 15122968 TI - Adolescents' attachment representations and developmental tasks in emerging adulthood. AB - The association between attachment representations and adolescents' coping with 3 developmental tasks of emerging adulthood-leaving home, advancing in the capacity for mature intimacy, and developing individuation-was examined. Israeli male adolescents (N = 88) were administered the Adult Attachment Interview during their high-school senior year. A year later, they and their friends reported on the adolescents' adjustment to mandatory military service. Three years later, participants and their parents reported on the adolescents' capacity for intimacy using an in-depth interview and on their individuation. An autonomous state of mind was associated with better coping with basic training and with a higher capacity for mature intimacy but was not associated with markers of individuation. The results highlight the importance of attachment representations in shaping an individual's developmental trajectory. PMID- 15122969 TI - Fetal neurobehavioral development: a tale of two cities. AB - Longitudinal neurobehavioral development was examined in 237 fetuses of low-risk pregnancies from 2 distinct populations--Baltimore, Maryland, and Lima. Peru--at 20, 24, 28, 32, 36, and 38 weeks gestation. Data were based on digitized Doppler based fetal heart rate (FHR) and fetal movement (FM). In both groups. FHR declined while variability, episodic accelerations, and FM-FHR coupling increased, with discontinuities evident between 28 and 32 weeks gestation. Fetuses in Lima had higher FHR and lower variability, accelerations, and FM-FHR coupling. Declines in trajectories were typically observed 1 month sooner in Lima, which magnified these disparities. Motor activity differences were less consistent. No sex differences in fetal neurobehaviors were detected. It is concluded that population factors can influence the developmental niche of the fetus. PMID- 15122970 TI - Audit of head injury management in Accident and Emergency at two hospitals: implications for NICE CT guidelines. AB - BACKGROUND: The National Institute for Clinical Excellence (NICE) has produced guidelines on the early management of head injury. This study audits the process of the management of patients with head injury presenting at Accident and Emergency (A&E) departments and examines the impact upon resources of introducing NICE guidelines for eligibility of a CT scan. METHODS: A retrospective audit of consecutive patients of any age, presenting at A&E with a complaint of head injury during one month in two northern District General Hospitals forming part of a single NHS Trust. RESULTS: 419 patients presented with a median age of 15.5 years, and 61% were male. 58% had a Glasgow Coma Score (GCS) recorded and 33 (8%) were admitted. Only four of the ten indicators for a CT scan were routinely assessed, but data were complete for only one (age), and largely absent for another (vomiting). Using just three (incomplete) indicators showed a likely 4 fold increase in the need for a CT scan. CONCLUSIONS: The majority of patients who present with a head injury to Accident and Emergency departments are discharged home. Current assessment processes and associated data collection routines do not provide the information necessary to implement NICE guidelines for CT brain scans. The development of such clinical audit systems in a busy A&E department is likely to require considerable investment in technology and/or staff. The resource implications for radiology are likely to be substantial. PMID- 15122971 TI - Incorrect identification of recent Asian strains of Coxsackievirus A16 as human enterovirus 71: improved primers for the specific detection of human enterovirus 71 by RT PCR. AB - BACKGROUND: Human enterovirus 71 has emerged as an important pathogen in the Asia Pacific region and it is important to be able to make a rapid and specific diagnosis for outbreak control. Recent Asian strains of Coxsackievirus A16 have changes in the VP1 gene which causes mispriming of widely used primers for human enterovirus 71 specific identification. METHODS: Local strains of Coxsackievirus A16 were sequenced in the VP4 and VP1 genes and using sequence alignment tools, an improved set of primers were designed for specific identification of human enterovirus 71. These primers were evaluated against virus isolates as well as primary clinical specimens. RESULTS: A total of 218 virus strains were tested. All 39 human enterovirus 71 isolates were positive and none of the 38 Coxsackievirus A16, 127 other enteroviruses and 14 prototype flaviviruses and adenoviruses were positive when tested with the new primers. When aliquots of primary specimens known to have yielded human enterovirus 71 were retrospectively tested, we found that within 2 months of collection of the specimens, greater than 90% were positive but that the success rate diminished rapidly to 18% after 2 years storage. CONCLUSIONS: Our new primers will be useful in rapid diagnosis of human enterovirus 71 infection, and can also be used as a screening tool in surveillance programmes for early warning of human enterovirus 71 transmission. PMID- 15122972 TI - Prolactin response to d-fenfluramine in combat-related post-traumatic stress disorder. AB - Central serotonergic function can be investigated by measuring the prolactin response to the serotonin releasing/uptake agent, d-fenfluramine. This study investigated the effect of diagnosis, depressive symptoms and history of alcohol or tobacco abuse or dependence on the d-fenfluramine test in combat-related post traumatic stress disorder (PTSD). Male, non-hospitalized combat-exposed veterans diagnosed with PTSD (DSM-III-R) and a similarly aged combat-exposed control group were assessed for both PTSD and depressive symptoms and prolactin responses to a 30-mg d-fenfluramine challenge test. Ninety-five subjects were studied; 23 were controls, 46 subjects met the criteria for current PTSD and 26 for past PTSD. There were no significant differences between the three groups for baseline prolactin, peak prolactin, and time to reach peak, delta prolactin or area under the curve of the prolactin vs. time curve. Depressive symptoms and history of alcohol or tobacco abuse or dependence did not have a confounding effect on the prolactin responses to d-fenfluramine. This study suggests that a blunted prolactin response to d-fenfluramine may be a consequence of combat exposure rather than PTSD. To confirm this, further studies involving both healthy and combat-exposed control groups in addition to subjects with PTSD of similar ages are required. PMID- 15122973 TI - Effect of treatment with bupropion on EEG sleep: relationship to antidepressant response. AB - The objective of this study was to assess whether treatment with sustained release bupropion (Wellbutrin-SR) produces alterations in electroencephalographic (EEG) sleep that are associated with clinical response to the drug. EEG sleep was measured in 20 patients with unipolar major depressive disorder before and after treatment with sustained-release bupropion. Each EEG sleep session consisted of two consecutive nights. Treatment with bupropion lasted for 8 wk. Compared to EEG sleep measures at baseline, treatment with bupropion significantly lengthened REM latency, increased REM activity and density during the first REM period, and increased total REM density. Differential response to treatment was associated with changes in REM activity and density, but not with REM latency. However, in contrast to many other antidepressants, REM sleep was not suppressed. PMID- 15122974 TI - Influence of immobilization stress on the levels of CaMKII and phospho-CaMKII in the rat hippocampus. AB - The phosphorylation of calcium/calmodulin-dependent protein kinase (CaMK) II, induced by an increase in the intracellular Ca2+ concentration, is involved in the alteration of brain functions such as memory formation. In the present study, we examined the influence of various immobilization stress paradigms on the phosphorylation of CaMKII (phospho-CaMKII) and CaMKII levels in the rat hippocampus. Immunoblot and immunohistochemical analyses were performed to examine the levels of CaMKII and phospho-CaMKII. Real-time quantitative polymerase chain reaction (PCR) was performed to analyse the mRNA levels of N methyl-D-aspartic acid (NMDA) and alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) receptor subtypes. Acute (single) and repeated (4 d), but not chronic (14 d), stress exposure of 45 min or longer duration significantly increased phospho-CaMKII levels without affecting the levels of CaMKII. Pre-treatment with NBQX, a selective AMPA receptor antagonist, significantly prevented this stress-induced increase. In contrast, two NMDA receptor antagonists, LY235959 and MK-801, showed no inhibitory effect on phospho CaMKII levels during acute stress. Neither acute nor chronic stress changed mRNA levels of NMDA and AMPA receptors. These results demonstrate that immobilization stress promotes the phosphorylation of CaMKII. The increase in the intracellular Ca2+ concentration by the activation of AMPA receptors may play a role in the stress-induced phospho-CaMKII in the rat hippocampus. PMID- 15122975 TI - Regulatory SNPs in complex diseases: their identification and functional validation. AB - Finding the genetic causes for complex diseases is a challenge. Expression studies have shown that the level of expression of many genes is altered in disease compared with normal conditions, but what lies behind these changes? Linkage studies provide hints as to where in the genome the genetic triggers--the mutations--might be located. Fine-mapping and association studies can give yet more information about which genes, and which changes in the genes, are involved in the disease. Recent examples show that single-nucleotide polymorphisms (SNPs), which are variations at the single-nucleotide level within an individual's DNA, in the regulatory regions of some genes constitute susceptibility factors in many complex diseases. This article discusses the nature of regulatory SNPs (rSNPs) and techniques for their functional validation, and looks towards what rSNPs can tell us about complex diseases. PMID- 15122976 TI - Hyponatremia of exercise. PMID- 15122977 TI - Brachial plexus injury in a male football player. PMID- 15122978 TI - Preparation and medical management of events in mountain and high-altitude environments. AB - Athletic pursuits in mountain and high-altitude environments continue to increase in popularity for the recreational and professional athlete. Accordingly, the sports medicine practitioner will have an increasing opportunity and responsibility to serve the unique needs of the mountain sport athlete. Many medical conditions occurring in the mountain and high-altitude environments will be unique to the Western medicine practitioner. This article discusses the medical care unique to athletes participating in these environments. PMID- 15122979 TI - Motorsports medicine. AB - Motorsports is the fastest growing professional sport in the United States. Each year approximately 14 drivers die, and many others are paralyzed or seriously injured. Although there is a common misconception that motorsports medicine is analogous to standard emergency or sports medicine, due to the unique racing environment a traditional approach to emergency medical services can be ineffective and may expose drivers, spectators, and medical personnel to great danger. This article is a general review of the evolving subspecialty of motorsports medicine. PMID- 15122980 TI - Safety, security, and preparing for disaster at sporting events. AB - There is a heightened awareness of terrorism in this country. There always remains the possibility of nonterrorist disasters at sporting venues. The team physician will be among the first medical responders to a disaster at a sporting venue. By being involved in the creation of an emergency action plan, learning the incident command system, understanding triage, and obtaining basic trauma life support skills, the team physician can be prepared to respond to mass casualty incidents at sporting events. PMID- 15122981 TI - Football: sideline management of injuries. AB - Football is reported to have one of the highest rates of injury among sports. Thus, it is becoming more commonplace for a physician to be present at the sideline to help manage injuries acutely. The team physician should be well equipped to provide game coverage by having the necessary equipment and knowledge of the injuries common in football. The physician should have an understanding of how to evaluate injuries at the sideline and when to send the player to the emergency room for further evaluation. PMID- 15122982 TI - Barriers and facilitators to sports medicine care. PMID- 15122983 TI - Cultural sensitivity in sports medicine. PMID- 15122984 TI - Knee pain in a recent immigrant. PMID- 15122985 TI - Ironman triathlon case history. PMID- 15122986 TI - How does medical insurance type influence sports medicine patient care? PMID- 15122987 TI - Training issues in elite young athletes. AB - Over the past decade, there has been a surge in the number of sports opportunities available to young athletes. Although physicians, parents, and coaches should promote healthy activity and participation, intense training at a young age can predispose exuberant young athletes to certain difficulties. Elite young athletes are at risk for overuse and growth plate injuries in certain sports. Intense training combined with inadequate nutrition may cause growth delay in elite young athletes, but this delay does not appear to affect permanent adult height. Weight training, when done properly, is safe and effective for prepubescent and pubescent athletes. Awareness of neurobehavioral development can help guide the process for appropriate sports participation. Young athletes should be closely monitored for signs of excessive physical and emotional stress so that sports participation can be fun and rewarding. PMID- 15122988 TI - Training principles for elite senior athletes. AB - Despite the passage of time, masters athletes are still capable of incredible performances. Nevertheless, overuse injuries are the most common challenge in this escalating cohort. The incidence of exertion-related cardiovascular events is also greater among older athletes, especially men, highlighting the importance of a thorough preparticipation medical evaluation before entry into senior athletic training programs and competition. Accordingly, masters athletes with a moderate to high risk for coronary artery disease, who desire to enter vigorous competitive events, should undergo peak or symptom-limited exercise testing. Training programs for senior athletes should be individually tailored and focus on the predominant energy pathways and performance requirements for a given sport. Although competition and "personal bests" are important, protection against sickness and disability are increasingly listed as the primary participative motives of older athletes. PMID- 15122989 TI - Oxidative damage to DNA, p53 gene expression and p53 protein level in the process of aging in rat brain. AB - Levels of 8-oxo2'dG (HPLC), p53 mRNA (PCR) and p53 protein (Western Blot) were estimated in four structures of rat brain, including grey matter (GM) of cerebral cortex, cerebral white matter (WM), cerebellum (C) and medulla oblongata (MO) of control (3.0-3.5-month-old) rats, 12- and 24-month-old rats. The level of oxidative DNA was statistically significantly higher in C of 24-month-old animals. Expression of p53 gene increased in C and also in the all other investigated brain parts, while the protein level of p53 was enhanced only in GM of 24-month-old rats. These data indicated that DNA oxidative damage and p53 gene expression increased significantly in aged brain. The higher expression of p53 gene in aged brain may suggest the activation of DNA repair processes. PMID- 15122990 TI - Ventilatory pattern and chemosensitivity in M1 and M3 muscarinic receptor knockout mice. AB - Acetylcholine (ACh) acting through muscarinic receptors is thought to be involved in the control of breathing, notably in central and peripheral chemosensory afferents and in regulations related to sleep-wake states. By using whole-body plethysmography, we compared baseline breathing at rest and ventilatory responses to acute exposure (5 min) to moderate hypoxia (10% O(2)) and hypercapnia (3 and 5% CO(2)) in mice lacking either the M(1) or the M(3) muscarinic receptor, and in wild-type matched controls. M(1) knockout mice showed normal minute ventilation (V(E)) but elevated tidal volume (V(T)) at rest, and normal chemosensory ventilatory responses to hypoxia and hypercapnia. M(3) knockout mice had elevated V(E) and V(T) at rest, a reduced V(T) response slope to hypercapnia, and blunted V(E) and frequency responses to hypoxia. The results suggest that M(1) and M(3) muscarinic receptors play significant roles in the regulation of tidal volume at rest and that the afferent pathway originating from peripheral chemoreceptors involves M(3) receptors. PMID- 15122991 TI - Dopamine1 receptor agonists reverse opioid respiratory network depression, increase CO2 reactivity. AB - In adult pentobarbital-anesthetized and unanesthetized decerebrate cats, the D(1)R agonists (6-chloro-APB, SKF-38393, dihydrexidine) given intravenously restored phrenic nerve and vagus nerve respiratory discharges and firing of bulbar post-inspiratory neurons after the discharges were abolished by the micro opioid receptor agonist fentanyl given intravenously. Reversal of opioid-mediated discharge depression was prevented by the D(1)R antagonist SCH23390. Iontophoresis of the micro-opioid receptor agonist DAMGO depressed firing of medullary bulbospinal inspiratory neurons. Co-iontophoresis of SKF-38393 did not restore firing and had no effect on bulbospinal inspiratory neuron discharges when applied alone. The D(1)R agonists given intravenously prolonged and intensified phrenic nerve and bulbospinal inspiratory neuron discharges. They also increased reactivity to CO(2) by lowering the phrenic nerve apnea threshold and shifting the phrenic nerve-CO(2) response curve to lower et(CO(2)) levels. Intravenous fentanyl on the other hand decreased CO(2) reactivity by shifting the phrenic nerve apnea threshold and the response curve to higher et(CO(2)) levels. Fentanyl effects on reactivity were partially reversed by D(1)R agonists. PMID- 15122992 TI - Fractal characteristics of breath to breath timing in sleeping infants. AB - We examined interbreath interval (IBI) time series of 19 term infants during active and quiet sleep for fractal properties using Fano factor analysis. For each time series we calculated the fractal exponent (alpha), comparing alpha for the original time series with two forms of surrogate data, a temporally independent surrogate set and an autoregressive surrogate set. alpha values were normally distributed between 0.79 and -0.22, and did not differ with sleep state. The fractal characteristics of the original time series were not retained in the temporally independent surrogate time series indicating that the distribution of intervals alone was not fractal, but were retained using autoregressive surrogates with an order of 10, suggesting that the fractal properties of the IBI time series were related to correlations between successive breaths. These observations suggest that some of the respiratory variability that occurs during sleep in infants, which in the past has been regarded as stochastic noise, may be the product of deterministic processes. PMID- 15122993 TI - Respiratory mechanics and pleural remodelling in pleurodesis induced by barium sulphate. AB - The aim of this study was to determine whether an intrapleural injection of barium sulphate would produce pleurodesis in rats. Additionally, respiratory mechanics and pleural remodelling were analysed. Single intrapleural injection of barium sulphate (100%) or saline was given to Wistar rats. Respiratory system, lung, and chest wall elastic, resistive and viscoelastic/inhomogeneous pressures were measured by the end-inflation occlusion method at 2 and 30 days after injection. The pleura were examined for gross and histopathological evidence of pleural inflammation and fibrosis, and the underlying lungs were also studied by morphometry. All pulmonary mechanical parameters increased at day 2, but were not different from control at 30 days after injection. Chest wall mechanical parameters did not change. Macroscopic evaluation demonstrated pleural adherence without haemothorax. Histopathologic analysis showed pleural inflammation and fibrosis. There was no alveolar inflammation or fibrosis in both groups. In conclusion, barium sulphate induced pleurodesis with either no changes in respiratory mechanics or lung lesion at day 30. PMID- 15122994 TI - Cardiac activity during airway resistance alterations with intravenous and inhaled methacholine. AB - Measurement of airway contractile responses to methacholine (MCh) is an important investigational tool in humans and mice. However, i.v. administration of MCh in murine models may suffer from potential cardiac sequelae produced by stimulation of cholinergic receptors within cardiac muscle. Therefore, we studied the i.v. (0 10,000 microg/kg) and aerosolized (inhaled; 0-25 mg/ml) administration of MCh, to determine their effects on pulmonary resistance (RL) and cardiac muscle activity (as heart rate; HR) in anesthetized, mechanically-ventilated C57Bl6 mice. MCh, i.v., increased RL but produced: (1) prolonged asystole (29-47 sec); (2) subsequent overshoot of baseline HR; and (3) prolonged HR recovery times (7-25 min), suggestive of sympathetic modulation after cholinergic stimulation. In contrast, inhaled MCh aerosol produced no change in HR, while increasing RL similar to i.v. MCh. These results suggest that, for specific instances utilizing the plethysmographic technique, inhaled MCh aerosol may be superior to i.v. administration, due to the avoidance of potential bouts of asystole that can confound experimental results and lead to premature death of mice. PMID- 15122995 TI - Improvement after lung volume reduction surgery: a role for inspiratory muscle adaptation. AB - In severe emphysema, lung volume reduction surgery (LVRS) can improve lung function and exercise tolerance. The maximal changes of forced expiratory volume in 1s (FEV(1)) and lung volume occur early after surgery, whereas maximal improvement of exercise tolerance occurs later. We tested the hypothesis that secondary adaptation of inspiratory muscles could explain this delayed clinical improvement. In that purpose, we evaluated nine consecutive patients before LVRS and up to 9 months post-operatively. Six weeks after LVRS, we observed an increase in FEV(1) and 6 min walk distance (6MWD). The gain in sniff nasal inspiratory pressure (SNIP) was inversely proportional to lung volume loss. Values of FEV(1) and lung volume were maintained throughout follow-up whereas SNIP values significantly increased from 6 weeks to 6 months post-LVRS. In the meantime, we observed an increase in 6MWD correlated with the SNIP increase. This suggests that in patients undergoing LVRS, early improvement of SNIP is proportional to decrease in lung volume whereas the further delayed improvement may be due, at least in part, to adaptation of the inspiratory muscles. PMID- 15122996 TI - A theoretical study of the effect of circadian rhythms on sleep-induced periodic breathing and apnoea. AB - This study employed a mathematical model of the respiratory control system to test the plausibility of the hypothesis that circadian rhythms in respiratory control can significantly influence respiratory stability at sleep onset. Computer simulations utilized a standardized "normal" sleep onset effect, superimposed upon systematic changes in chemoreflex parameters that mimicked the peaks and troughs of normal and high amplitude circadian rhythms. The analysis predicted that circadian influences may augment sleep-induced periodic breathing in nocturnal sleep compared with daytime naps. Furthermore, increased circadian amplitude of chemoreflex threshold, or absence of a circadian rhythm in peripheral chemosensitivity, each acted to stabilize respiration during daytime sleep onset and promote periodic breathing during nocturnal sleep onset. High amplitude circadian rhythms in respiratory control were predicted to cause an increasing number and duration of obstructive apnoeas from early to late night. It is suggested that the circadian timing system creates a nocturnal window of respiratory vulnerability and that abnormal circadian rhythms could potentially induce nocturnal sleep apnoea, even in individuals with normal sleep mechanisms. PMID- 15122997 TI - Ion transport across Xenopus alveolar epithelium is regulated by extracellular ATP, UTP and adenosine. AB - Native alveolar epithelium from Xenopus lung was used for electrophysiological Ussing chamber experiments to investigate ion transport regulation. The tissue exhibits a considerable absorption of Na(+) ions and this transepithelial transport is largely up-regulated after treatment of donor animals with ACTH. Extracellular ATP, UTP and adenosine were tested for their regulating effects and all three increased I(sc), which was mainly due to a stimulation of amiloride sensitive Na(+) transport (increase of I(ami) 32% for ATP, 21% for UTP, 25% for adenosine). Solely the effect of UTP was completely abolished in the presence of amiloride. In contrast, the effects of ATP or adenosine disappeared under Cl(-) free conditions. ATP and UTP proved to have additive effects and pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS), an antagonist of purinergic receptors, inhibited selectively the effect of UTP on I(sc). Further, I(sc) was increased by the P2X selective agonist beta,gamma-meATP. We were able to demonstrate, that extracellular purines and pyrimidines play a possible role as auto/paracrine messengers for alveolar ion transport regulation in Xenopus lung. PMID- 15122998 TI - Apoptotic proteins in the course of aging of central nervous system in the rat. AB - Studies were performed on the level of cells with damaged DNA (TUNEL), the level of protein engaged in DNA repair (PARP) and the level of proteins indicating the extent of apoptosis (Bax:Bcl-2) (Western blot). The studies were performed on cerebral cortex (GM), white matter (WM), medulla oblongata (MO), cerebellum (C) of rats, 3.0-3.5-, 12-, 24-months of age. The highest levels of DNA injury in GM of 1-year-old rats and in MO of 2-year-old rats were accompanied by peak levels of PARP. In the remaining structures (WM, C) levels of DNA injury showed no correspondence with levels of PARP. Levels of Bax proteins exceeded levels of Blc 2 protein in all cerebral structures of young rats. In old animals, Bax protein continued to exceed Blc-2 levels both in GM and in MO, in which most pronounced fragmentation of DNA was observed. The data indicated that in spite of high level of TUNEL positive cells in aged brain PARP and Bcl-2 are probably engaged in protection of the cells against death. PMID- 15122999 TI - Role of L-glutamate in the locus coeruleus of rats in hypoxia-induced hyperventilation and anapyrexia. AB - Locus coeruleus (LC) is a noradrenergic nucleus in the pons which has been reported to play an inhibitory role in the ventilatory response to hypoxia. Since LC contains glutamatergic receptors and L-glutamate is known to participate in the ventilatory and thermoregulatory responses to hypoxia, the effects of kynurenic acid (KYN, a glutamatergic receptor antagonist) microinjected into the LC in the hypoxic hyperventilation and anapyrexia (a regulated drop in body temperature [Tb]) were examined. Ventilation (V) and Tb were measured before and after a microinjection of KYN (10 nmol/0.1 microl) into the LC, followed by hypoxia. Control rats received a saline injection. Under normoxia, KYN treatment did not affect V or Tb. Typical hypoxia-induced hyperventilation and anapyrexia were observed after saline injection. KYN injection caused an increase in the ventilatory response, acting on tidal volume (Vt), but did not affect the anapyrexic response to hypoxia. These data suggest that L-glutamate in the LC is an excitatory neurotransmitter that activates an inhibitory pathway to reduce the hypoxic ventilatory response, similarly to the data reported for rostral ventrolateral medulla (VLM). The role of L-glutamate into the LC and VLM opposes its effect on other nuclei such as the nucleus of the solitary tract and ventromedullary surface, where the neurotransmitter participates in an excitatory pathway of the ventilatory response. PMID- 15123000 TI - Pulmonary mechanics and lung histology in acute lung injury induced by Bothrops jararaca venom. AB - Pulmonary mechanics [static (Est) and dynamic (Edyn) elastances, resistive (DeltaP1) and viscoelastic pressures (DeltaP2)], histology, and bronchoalveolar lavage fluid (BALF) from BALB/c mice were analysed 1, 24, 48 and 72 h after intravenous injection of saline or Bothrops jararaca crude venom [0.3 (V0.3) or 1 (V1) microg.g(-1)]. Est, Edyn, and DeltaP2 increased at 1 h in both V groups, being significantly higher in V1 than in V0.3, decreasing progressively, reaching control values at 48 h in V0.3, but remaining altered in V1 at 72 h. DeltaP1 augmented in V1 at 1 h, returning to normal at 72 h. Histological changes in V0.3 group included interstitial oedema, alveolar collapse, and increased cellularity, which returned to normal at 48 h. These changes were more intense in V1 group, with alveolar oedema and haemorrhage. BALF showed time-dependent neutrophil influx in V0.3. In conclusion, venom led to time- and dose-dependent pulmonary mechanical changes, together with moderate inflammation in V0.3 and acute lung injury in V1. PMID- 15123001 TI - Branching properties of the pulmonary arterial tree during pre- and postnatal development. AB - We measured arterial diameter as a function of generation number (#) in the arteriograms of six postmortem lung preparations from human infants aged 35 to 48 wks post-conceptional age (PCA). The log-log plot of mean diameter as a function of generation number revealed a linear decrease in mean and median diameter with increasing #, which was characterized by its slope alpha and intercept beta (linear regression). The mean arterial diameter per generation as well as the ratio (F) between mother and larger daughter branch and the relative ratio of asymmetry (A) between the larger and the smaller daughter branch was calculated. The values of F(#) and A(#) were found to be constant between 2 and 15 generations in individual lungs which is consistent with the pulmonary arterial tree exhibiting fractal properties. The averaged values (F and A within a subject) of F(#) and A(#) as well as alpha and beta were determined for each lung preparation and found to be constant from 35 to 48 weeks of age, revealing unchanged branching properties in the pre- and postnatal phase of human lung development. PMID- 15123002 TI - Basic features of hypoxic pulmonary vasoconstriction in mice. AB - Hypoxic pulmonary vasoconstriction (HPV) matches lung perfusion with ventilation which tends to optimize pulmonary gas exchange. Investigations using genetically engineered mice represent a promising approach to understand the underlying mechanisms. Our goal was to characterize basic features of HPV in the isolated buffer-perfused and ventilated mouse lung system. HPV was reproducible for several hours when ventilating the lungs with 1% O2 (10 min) alternated with normoxic ventilation periods (21% O2, 15 min). HPV was well elicitable and most constant using Krebs-Henseleit buffer with the addition of hydroxyethylamylopectin as an oncotic agent. Inhibition of both lung NO and prostanoid formation amplified HPV in an over-additive fashion. HPV was higher in BALB/c mive as compared to C57BL/6 mice, and was approximately threefold enhanced under positive pressure ventilation as compared to negative pressure ventilation. A three hour hypoxic ventilation period resulted in a biphasic vasoconstrictor response with loss of posthypoxic vasodilatation. In summary, we have characterised HPV and established an experimental set-up optimized for investigation of the basic mechanisms of HPV in mice. PMID- 15123003 TI - Oxygen uptake kinetics during severe exercise: a comparison between young and older men. AB - This study examined the relationship between the slow component of oxygen uptake (VO2) kinetics and muscle electromyography (EMG) during severe exercise in nine young (21.7+/-0.9 yr) and nine older (71.6+/-0.8 yr) men. Oxygen uptake (VO2) and surface EMG activity of the left vastus lateralis muscle were measured during a 7 min square-wave bout of severe exercise on a cycle ergometer. The absolute amplitude of the VO2 slow component was greater and occurred approximately 60 s earlier in the young compared to older subjects. However, the rate of increase in the slow component, expressed as a percentage of the total VO2 response per unit time, was not different between young and older subjects (young: 4.8+/-0.5%.min( 1); older: 4.9+/-0.6%.min(-1)). The mean power frequency (MPF) of the EMG increased significantly during the slow component phase of exercise by 6.4+/-1.0% in the young and by 5.4+/-0.7% in the older group and this rise was not significantly different between the two groups. These results indicate that normal ageing may not alter the VO2 slow component (measured as the rate of increase in VO2) and that this finding may be related to similar muscle fibre recruitment patterns in the two groups during severe-intensity exercise. PMID- 15123004 TI - Oral airway resistance during wakefulness in eucapnic and hypercapnic sleep apnea syndrome. AB - The purpose of this study was to evaluate whether there was an abnormal increase of upper airway resistance in the sitting and supine positions in hypercapnic obstructive sleep apnea syndrome (OSAS) patients compared with eucapnic OSAS or normal controls as measured by impulse oscillometry (IOS) while awake. Twenty subjects without OSAS served as controls (group I), and 20 patients with moderate or severe eucapnic OSAS (group II) and another eight hypercapnic severe OSAS patients (group III) were studied. Group II was further divided into two subgroups. Group IIa consisted of 14 subjects whose BMI was less than 35 and group IIb of six subjects whose BMI was greater than 35. All subjects also had an overnight sleep study. Oral airway resistance (AR) (including impedance (Zrs), resistance (R) and reactance (X)) was measured by impulse oscillometry (IOS) (MasterScreen IOS, VIASYS Healthcare GmbH, Germany) in the upright (seated) position and then in the supine position while awake. The results demonstrated that in both group I and group II, Zrs was normal in the sitting position. However, there was a high Zrs in the supine position for group II patients. In contrast, in group III patients, there was a high Zrs in both the sitting and supine positions. In conclusion, upper airway resistance was increased both sitting and supine in the hypercapnic OSAS patients; this would presumably increase the work of breathing and might explain why these subjects were hypercapnic while awake, while eucapnic OSAS patients and normal controls were not. Secondly, the increased upper airway resistance in the supine position in the eucapnic OSAS patients may contribute to their OSAS. PMID- 15123006 TI - Pathophysiology of bone loss in patients receiving anticonvulsant therapy. AB - Many studies have shown that patients taking antiepileptic drugs (AEDs) are at increased risk for metabolic bone disease and low bone mineral density. Although early reports of bone disease in patients with epilepsy often involved institutionalized patients, who may be at risk because of lack of physical activity, reduced sunlight exposure, and poor nutrition, low bone density has also been reported in well-nourished, ambulatory outpatients with epilepsy. Traditionally, attention to the problem of AED-induced bone loss has been focused on those drugs that induce the hepatic cytochrome P450 enzyme system, thereby increasing the metabolism of vitamin D. However, the mechanisms of AED-induced bone loss appear to be multiple, and all types of AEDs are potentially implicated. Besides hepatic enzyme induction, mechanisms may include direct effects of AEDs on bone cells, resistance to parathyroid hormone, inhibition of calcitonin secretion, and impaired calcium absorption. An understanding of bone biology and the pathophysiology of bone loss can aid in the identification and monitoring of patients at risk and in the planning of appropriate prophylactic and therapeutic measures, by which most of the morbidity associated with AED induced bone loss can be prevented. PMID- 15123007 TI - Evaluation and measurement of bone mass. AB - Patients taking antiepileptic drugs (AEDs) have an increased risk for osteoporosis and osteoporotic fractures. Bone mineral density (BMD) is the best predictor of fracture risk. Measurement of BMD allows the identification of patients at risk before they suffer a fracture. The current "gold standard" for the measurement of BMD and diagnosis of osteopenia and osteoporosis is dual energy X-ray absorptiometry. Clinicians should be familiar with the ways in which bone mass measurements are reported and should understand how to use BMD measurements in making treatment decisions and monitoring treatment. They should also be aware of current controversies, including the role of peripheral versus central BMD measurements and the use of different reference databases for different patient groups. Laboratory assessment to identify secondary causes of osteoporosis, such as low vitamin D status in patients taking AEDs, is useful, but the role of markers of bone turnover in clinical practice has not been well defined. PMID- 15123008 TI - Epilepsy and bone health in adults. AB - Adults taking antiepileptic drugs (AEDs) have an augmented risk for osteopenia and osteoporosis because of abnormalities of bone metabolism associated with AEDs. The increased fracture rates that have been described among patients with epilepsy may be related both to seizures and to AEDs. The hepatic enzyme-inducing AEDs phenytoin, phenobarbital, and primidone have the clearest association with decreased bone mineral density (BMD). Carbamazepine, also an enzyme-inducing drug, and valproate, an enzyme inhibitor, may also adversely affect bone, but further study is needed. Little information is available about specific effects of newer AEDs on bone. Physicians are insufficiently aware of the association between AEDs and bone disease; a survey found that fewer than one-third of neurologists routinely evaluated AED-treated patients for bone disease, and fewer than 10% prescribed prophylactic calcium and vitamin D. Physicians should counsel patients taking AEDs about good bone health practices, and evaluation of bone health by measuring BMD is warranted after 5 years of AED treatment or before treatment in postmenopausal women. PMID- 15123009 TI - Bone health in pediatric epilepsy. AB - Childhood and adolescence are critical periods of skeletal mineralization. Peak bone mineral density achieved by the end of adolescence determines the risk for later pathological fractures and osteoporosis. Chronic disease and medication often adversely affect bone health. Epilepsy is one of the most common neurological conditions occurring in persons under the age of 21. Epilepsy may affect bone in a number of ways. Restrictions of physical activity imposed by seizures; limitations on physical activity resulting from cerebral palsy, frequently present in patients with symptomatic epilepsy; and medications used to treat seizures can all adversely affect bone health. It has long been observed that treatment with phenytoin and phenobarbital can be associated with rickets. More recently, established agents such as carbamazepine and valproate have been shown to be associated with a lowering of bone mineral density. The literature related to bone health in pediatric epilepsy is reviewed, although it should be noted that these data are limited. PMID- 15123010 TI - Fractures, epilepsy, and antiepileptic drugs. AB - The risk for skeletal fractures in patients with epilepsy is two to six times greater than in the general population. Fractures may be caused by seizures themselves or by falls, with or without seizures. Side effects of antiepileptic drugs (AEDs), such as ataxia, and coexisting neurological deficits contribute to the risk for falls. The effects of older AEDs on bone mineral density probably increase the risk for fractures associated with seizures and falls. Preventive measures include optimal control of seizures and supplementation with calcium and vitamin D. Whether newer AEDs prove to be without adverse effects on bone mineral metabolism remains to be determined. PMID- 15123011 TI - Treatment of anticonvulsant drug-induced bone disease. AB - Although the pathophysiology of bone disease in patients treated with anticonvulsant drugs may vary, most affected patients have increased bone remodeling rather than decreased mineralization. Milder cases may show high bone turnover without significant loss of cortical or trabecular bone. Cases of intermediate severity may exhibit the characteristic features of a high-turnover osteopenia/osteoporosis, but some patients with severe bone disease may manifest the features of an osteomalacic disorder. Prophylactic vitamin D supplementation at doses up to 2000 IU/day can be recommended for all patients on initiation of anticonvulsant therapy. A calcium intake of 600-1000 mg/day should also be ensured. If an osteopenic/osteoporotic disorder exists, treatment with 2000-4000 IU/day vitamin D is appropriate. Vitamin D doses of 5000-15,000 IU/day may be needed to treat osteomalacia. Conventional treatment with bisphosphonates may be needed when the response to vitamin D is inadequate. However, routine use of bisphosphonates in patients receiving long-term anticonvulsant therapy cannot at present be recommended. PMID- 15123013 TI - Accuracy of self-reported neuropsychological functioning in individuals with epileptic or psychogenic nonepileptic seizures. AB - The aim of this study was to determine the degree to which subjective ratings of neurocognitive ability accurately reflect objectively measured neuropsychological functioning in patients diagnosed with epileptic (ES, n = 45) or psychogenic nonepileptic (PNES; n = 37) seizures. Patients received a battery of neuropsychological tests, measures of current mood state, and the Quality of Life In Epilepsy-89 questionnaire. Results indicated that subjective ratings of neuropsychological functioning were only partially accurate within each group. Patients with ES accurately rated their memory function, but overestimated language and attention abilities. Patients with PNES accurately rated attention, but underestimated memory and overestimated language. In both groups, poorer self reported neurocognitive functioning was strongly related to poorer mood state; however, mood state did not predict objectively measured neurocognitive abilities. Given the inaccuracies that exist in patient self-report, results highlight the importance of a comprehensive neuropsychological assessment when evaluating the neurocognitive status of individuals with seizures. PMID- 15123014 TI - Induction of audiogenic seizures in imipenem/cilastatin-treated rats. AB - We investigated the effect of intense audiogenic stimulation (AGS) on rats treated with the antibiotic imipenem and dipeptidase inhibitor cilastatin (Imi/Cil). Under pentobarbital anesthesia (40 mg/kg) adult male Wistar rats were implanted with electrodes and cannulas were placed in the right lateral ventricle. Animals were divided into the following groups: (1) vehicle, (2) Imi/Cil 10 microg/10 microg, (3) Imi/Cil 25 microg/25 microg, (4) vehicle+AGS, (5) Imi/Cil 10 microg/10 microg +AGS, and (6) Imi/Cil 25 microg/25 microg +AGS. Imi/Cil was administered intracerebroventricularly in 5 microl of physiological saline. AGS (100+/-3 dB, 60 seconds) was applied at 15-minute intervals after the injection. Imi/Cil-induced seizures (twitching, forelimb clonus, headnodding, rearing, and clonic convulsions) and Imi/Cil-audio-induced seizures (wild running, clonic and tonic convulsions) were scored according to appropriate rating scales. Imi/Cil provoked convulsions dose-dependently. Each behavioral seizure response had a characteristic EEG correlate. AGS by itself did not provoke seizures in untreated rats. Sound stimulation in Imi/Cil-injected rats elicited typical audiogenic seizures, which were induced during five AGS tests (75 minutes postinjection). In most cases audiogenic seizures were not associated with epileptiform activity in the EEG, indicating that spreading of seizures did not involve the cortex. Since Imi/Cil-induced and Imi/Cil-audio-induced seizures differed behaviorally and electroencephalographically, it is suggested that different neural pathways are responsible for these two types of seizures: neuronal networks in the cortex are involved in Imi/Cil-induced seizures, whereas audiogenic seizures use networks residing primarily in the brainstem. PMID- 15123015 TI - Frequency of EEG abnormalities in age-matched siblings of autistic children with abnormal sleep EEG patterns. AB - Epileptiform activity in sleep has been described even in the absence of clinical seizures in 43-68% of patients with autistic spectrum disorders (ASDs). Genetic factors may play a significant role in the frequency of epilepsy, yet the frequency in normal age-matched controls is unknown. We studied overnight ambulatory electroencephalograms (EEGs) in 12 nonepileptic, nonautistic children with a sibling with both ASDs and an abnormal EEG. EEG studies were read and described independently by two pediatric epileptologists; 10 were normal studies and 2 were abnormal. The occurrence of abnormal EEGs in our sample (16.6%) was lower than the reported occurrence in children with ASDs. Further, the two abnormal EEGs were of types typically found in childhood and were different from those found in the ASD-affected siblings. The lack of similarity between sibling EEGs suggests that genetic factors alone do not explain the higher frequency of EEG abnormalities reported in ASDs. PMID- 15123016 TI - Oxcarbazepine in pregnancy: clinical experience in Argentina. AB - The potential teratogenicity of antiepileptic drugs (AEDs) is a major concern for women with epilepsy who are considering pregnancy. Traditional AEDs are associated with an at least twofold risk of fetal malformations compared with the general population. The risk of malformations with newer AEDs is unclear. This article reports the multicenter clinical experience in Argentina of pregnant women with epilepsy receiving AEDs. Of 114 pregnancies monitored, 16 newborns had anomalies: 3 cardiac, 3 skull, and 2 gastrointestinal malformations, and 8 facial dysmorphies. Most fetal anomalies were observed following exposure to phenobarbital, valproate, and carbamazepine. Of 55 babies exposed to the new generation AED oxcarbazepine (20 as combination therapy and 35 as monotherapy), one malformation (cardiac) was reported (in a patient receiving oxcarbazepine and phenobarbital). Thus, newer AEDs may have a lower teratogenic risk than traditional AEDs. These data add to the growing experience with AED therapy in pregnant women with epilepsy. PMID- 15123017 TI - Effects of nimodipine on the behavioral sequalae of experimental status epilepticus in prepubescent rats. AB - OBJECTIVE: The goal of this study was to investigate the potential protective effects of nimodipine (ND), a calcium channel blocker, on the acute manifestations and long-term behavioral sequalae of experimental status epilepticus (SE). METHODS: Three groups of Postnatal Day (P) 35 rats undergoing kainic acid (KA)-induced SE were injected with phenobarbital (PB) and/or ND, and were subsequently compared with rats injected with KA alone and normal control rats. Behavioral parameters were assessed by the Morris water maze, open field, and handling tests at P125-P135. Acute seizures and spontaneous recurrent seizures (SRS) were assessed by videotape techniques. RESULTS: PB reduced the severity of SE acutely, and protected completely against subsequent long-term SRS, memory impairment, and hyperactivity, and partially against aggressivity. ND alone had no effect on acute seizure activity, but did protect against subsequent SRS and memory impairment, and partially against aggressivity. When administered together, PB and ND had effects similar to those seen with PB alone. However, in addition, and unlike the PB- and ND-alone groups, the PB-ND group was completely protected against KA-induced increased aggressivity. CONCLUSIONS: Activation of L type calcium channels contributes to the long-term behavioral sequalae of KA induced SE, but is not essential for the development and maintenance of SE. ND has protective effects in SE when given alone or in conjunction with a traditional antiepileptic drug. Calcium channel blockers should be further investigated as add-on protective agents in models of SE and possibly in clinical trials. PMID- 15123018 TI - Use of the Brief Visuospatial Memory Test-Revised (BVMT-R) in neuropsychological evaluation of epilepsy surgery candidates. AB - Many commonly used measures of figural reproduction have been found to be relatively insensitive to the effects of right temporal lobe dysfunction. More positive findings are present in studies using measures of figural learning over repeated trials. This study examined the use of a commercially available measure of figural learning in a sample of epilepsy patients undergoing presurgical neuropsychological testing. The Brief Visuospatial Memory Test-Revised (BVMT-R) was administered to 47 subjects undergoing comprehensive presurgical workups with video and EEG monitoring. Groups with left (N = 25) and right (N = 22) temporal lobe seizures did not differ on BVMT-R measures of learning, delayed recall, or yes/no recognition. Approximately 29% of the sample exhibited "impaired" performance in comparison to published norms. The results of receiver operating curve analyses revealed little discrimination in performance between groups. The BVMT-R does not appear to have the sensitivity required for assessing nonverbal memory in this population. PMID- 15123019 TI - Long-term behavioral and morphological consequences of nonconvulsive status epilepticus in rats. AB - The aims of the present study were to ascertain whether nonconvulsive status epilepticus (NCSE) could give rise to long-term behavioral deficits and permanent brain damage. Two months after NCSE was elicited with pilocarpine (15 mg/kg i.p.) in LiCl-pretreated adult male rats, animals were assigned to either behavioral (spontaneous behavior, social interaction, elevated plus-maze, rotorod, and bar holding tests) or EEG studies. Another group of animals was sacrificed and their brains were processed for Nissl and Timm staining as well as for parvalbumin and calbindin immunohistochemistry. Behavioral analysis revealed motor deficits (shorter latencies to fall from rotorod as well as from bar) and disturbances in the social behavior of experimental animals (decreased interest in juvenile conspecific). EEGs showed no apparent abnormalities. Quantification of immunohistochemically stained sections revealed decreased amounts of parvalbumin- and calbindin-immunoreactive neurons in the motor cortex and of parvalbumin positive neurons in the dentate gyrus. Despite relatively inconspicuous manifestations, NCSE may represent a risk for long-term deficits. PMID- 15123020 TI - Time to treatment in prolonged seizure episodes. AB - Prompt intervention in seizure emergencies is critical to reducing morbidity and mortality risks associated with status epilepticus. To determine the need for wider education about the benefits of at-home treatment, we examined the time from seizure onset to initial treatment in a cohort of patients with epileptic seizures. The seizure database of patients admitted in the greater Richmond, Virginia, area during a 5-year period (1989-1994) was queried to extract time to seizure treatment. Records were available for 889 patients. Patients were divided into two subgroups: children (age < 16 years, 29.7% of the cohort) and adults. Time to seizure treatment varied broadly; only 41.5% of all patients received their first antiepilepsy drug within 30 minutes. Time to treatment did not significantly differ between age groups. This baseline study supports the need for patient education regarding seizure emergencies and wider availability of at home treatment options to shorten time to seizure treatment. PMID- 15123021 TI - Effects of topiramate on memory performance on the intracarotid amobarbital (Wada) test. AB - We evaluated the effects of topiramate (TPM) on memory function in the intracarotid amobarbital (Wada) test in nine patients with mesial temporal lobe epilepsy (MTLE) whose antiepileptic drugs (AEDs) included TPM and compared their scores with those of 16 patients with MTLE on AEDs not including TPM. Sodium amobarbital was injected first into the hemisphere ipsilateral to the seizure focus and then into a contralateral site, and the patients were tested for naming and memorization. There was no statistical difference in percentage memory scores between the two patient groups following the contralateral injection. After the ipsilateral injection, however, TPM patients had significantly lower percentage memory scores compared with non-TPM patients (P < 0.02). We conclude that a possible adverse effect of TPM on memory performance should be considered when evaluating the Wada test memory scores of patients on TPM therapy. PMID- 15123022 TI - Levetiracetam prevents changes in levels of brain-derived neurotrophic factor and neuropeptide Y mRNA and of Y1- and Y5-like receptors in the hippocampus of rats undergoing amygdala kindling: implications for antiepileptogenic and mood stabilizing properties. AB - The amygdala-kindling model has been proposed as a model of sensitization processes with relevance to epilepsy as well as affective disorders. Levetiracetam is a novel anticonvulsant drug that delays the process of kindling, i.e., possesses antiepileptogenic properties. Preliminary reports also suggest a mood-stabilizing potential for levetiracetam. Brain-derived neurotrophic factor (BDNF) and neuropeptide Y (NPY) are central modulators of seizure activity, which undergo plastic changes during kindling epileptogenesis. Consequently, we investigated the regulation of BDNF and NPY mRNA and Y1-, Y2-, and Y5-like receptor binding in the hippocampus of vehicle-pretreated, partially and fully amygdala-kindled rats and corresponding levetiracetam-pretreated rats (40 mg/kg i.p.). The present data indicate that the process of kindling is associated with an upregulation of hippocampal BDNF and NPY mRNA levels and downregulation of Y1- and particularly Y5-like receptors. Pretreatment with levetiracetam markedly delays the progression of kindling and, in addition, exhibits a clear anticonvulsant effect. These effects are associated with abolition of the kindling-induced rise in BDNF and NPY mRNA and increasing levels of Y1- and particularly Y5-like receptors in all hippocampal subfields. Lastly, the present study reveals that an identical dose of levetiracetam reduced immobility in the rat forced swim test, the first experimental evidence indicative of an antidepressant and/or mood stabilizer-like profile of this drug. Considering that animal depression models display impairments in hippocampal NPY systems that become normalized following mood-stabilizing treatment, and that exogenous NPY exerts anticonvulsant as well as antidepressive-like activity in rodents, it is a heuristic possibility that increased hippocampal excitability and affective symptomatology may converge on an impaired hippocampal NPY function. Speculatively, the ability of levetiracetam to increase hippocampal Y1- and Y5 like receptor levels may have implications for the antiepileptic properties of levetiracetam, as well as its purported mood-stabilizing properties. PMID- 15123023 TI - Clinical efficacy of galvanic skin response biofeedback training in reducing seizures in adult epilepsy: a preliminary randomized controlled study. AB - We investigated the effect of galvanic skin response (GSR) biofeedback training on seizure frequency in patients with treatment-resistant epilepsy. Eighteen patients with drug-refractory epilepsy were randomly assigned either to an active GSR biofeedback group (n = 10) or to a sham control biofeedback group (n = 8). Biofeedback training significantly reduced seizure frequency in the active biofeedback group (P = 0.017), but not the control group (P > 0.10). This was manifest as a significant between-group difference in seizure reduction (P 0.01). Furthermore, there was a correlation between degree of improvement in biofeedback performance and reduction of seizure frequency (rho = 0.736, P = 0.001), confirming that the effect of biofeedback treatment was related to physiological change. Our findings highlight the potential therapeutic value of GSR biofeedback in reducing seizure frequency in patients with drug-resistant epilepsy. PMID- 15123024 TI - Quality of life improvement with conversion to lamotrigine monotherapy. AB - This report describes the effect on patient-reported quality of life (QOL) after reduction from two drugs to monotherapy with lamotrigine. Patients taking lamotrigine (LTG) with an enzyme-inducing drug were converted to LTG monotherapy for a 12-week follow-up. Changes in QOLIE-31 between baseline and follow-up were compared with physicians' global change ratings and patient-reported health status. Total QOLIE-31 scores increased 10.7 points for patients rated by physicians as having mild improvement, and 17 points for those reported as having moderate to marked improvement. Subscale scores also increased by minimum important change (MIC) amounts (> or = 11.76), with the largest change in Cognition, Energy, Medication Effects, and Seizure Worry subscales. The data also support > or = 11 MIC as a clinically important change in total score for the QOLIE-31. Exploratory analyses also provide information about MIC for individual subscales (8-18 for physician rated global change, 10-26 for patient-rated global health status change). This study demonstrates the value of reduction to monotherapy from the patients' and physicians' perspectives. PMID- 15123025 TI - Levetiracetam in adult patients with and without learning disability: focus on behavioral adverse effects. AB - Optimal antiepileptic drug treatment in patients with learning disability (LD) represents a particular challenge. These patients are often unable to report toxicity, and side effects may manifest as behavioral problems. The aim of this open study was to compare efficacy and tolerability of levetiracetam (LEV) in patients with LD and those without LD. One hundred eighty-four consecutive adult patients who received LEV were followed for an average of 8.1 months. Fifty-six patients (30%) had LD. Thirty-nine percent of patients with refractory epilepsy (37% with and 40% without LD) had > 50% seizure reduction. Significantly more behavioral side effects (23% vs 10%) and a tendency toward less reported somatic central nervous side effects were found in the LD group. We conclude that LEV is equally effective and well tolerated in both patients with LD and patients without LD. However, behavioral problems are more frequent in patients with LD, whereas the tendency toward seizure increase is not enhanced. PMID- 15123026 TI - Warrington's recognition memory for faces: interpretive strategy and diagnostic utility in temporal lobe epilepsy. AB - Performance on Warrington's Recognition Memory Test for Faces (RMF) is thought to rely largely on the integrity of the right temporal lobe. Epilepsy research, however, has been mixed with respect to the diagnostic utility of the RMF. Based on the psychometric properties of the RMF, we investigated the moderating role of intelligence on test classification accuracy in 53 presurgical patients with temporal lobe epilepsy. Classification accuracy rates were poor for the entire sample, but when the sample was divided based on IQ, classification statistics showed a strong degree of diagnostic utility for the RMF among patients with lower IQ levels versus those with higher IQ levels. These findings support the diagnostic utility of the RMF in epilepsy and underscore the moderating role of intelligence on RMF performance. PMID- 15123027 TI - High-resolution functional MRI at 3T in healthy and epilepsy subjects: hippocampal activation with picture encoding task. AB - Functional MRI (fMRI) studies of memory with coarse resolution of 4 x 4 x 5 mm often fail to demonstrate blood oxygenation level-dependent (BOLD) activation in the hippocampal formation. This failure occurs when nonactivating white matter is averaged with the signal from hippocampal gray matter, attenuating the total BOLD signal from a single voxel due to the "partial volume effect." In this study, we evaluated the suitability of high-resolution fMRI at 3T (voxel size 2 x 2 x 3 mm) for improved visualization of hippocampal activation during memory encoding in 21 healthy and 6 epilepsy subjects. We used a picture encoding task (block design) that involved memorization of indoor and outdoor scenes along with an appropriate resting task. Region-of-interest (ROI) analysis was performed; laterality indices (LIs) were calculated based on hippocampal ROIs (hROIs) or on global medial temporal ROIs (mtROIs). In 19 healthy subjects, robust bilateral BOLD signal changes within both ROIs were noted. The mean LI+/-SD for the hROIs is -0.12+/ 0.06 and that for the medial temporal ROIs -0.12+/-0.05, with correlation between the LIs (r = 0.59, P = 0.009). Good concordance was noted between the surgical outcome and memory lateralization with the fMRI task employed in this study. The preliminary results are encouraging, and with continuing improvements in MRI scanner technology, we expect fMRI of the hippocampal formation at higher resolution to be possible and preferable. Furthermore, these results suggest that a larger study to test the utility of high-resolution fMRI in epilepsy presurgical evaluation is needed. PMID- 15123028 TI - Comparison of intranasal midazolam with intravenous diazepam for treating acute seizures in children. AB - Midazolam, a water-soluble benzodiazepine, is usually given intravenously in status epilepticus. The aim of this study was to determine whether intranasal midazolam is as safe and effective as intravenous diazepam in the treatment of acute childhood seizures. Seventy children aged 2 months to 15 years with acute seizures (febrile or afebrile) admitted to the pediatric emergency department of a general hospital during a 14-month period were eligible for inclusion. Intranasal midazolam 0.2 mg/kg and intravenous diazepam 0.2 mg/kg were administered after intravenous lines were established. Intranasal midazolam and intravenous diazepam were equally effective. The mean time to control of seizures was 3.58 (SD 1.68) minutes in the midazolam group and 2.94 (SD 2.62) in the diazepam group, not counting the time required to insert the intravenous line. No significant side effects were observed in either group. Although intranasal midazolam was as safe and effective as diazepam, seizures were controlled more quickly with intravenous diazepam than with intranasal midazolam. Intranasal midazolam can possibly be used not only in medical centers, but also in general practice and at home after appropriate instructions are given to families of children with recurrent seizures. PMID- 15123029 TI - Improvement in alertness and behavior in children treated with combination topiramate and vagus nerve stimulation. AB - It has been reported that vagus nerve stimulation (VNS) improves behavior in children, whereas topiramate has a less clear effect. Three boys, aged 5-12 years, with generalized slow spike-wave discharges and refractory epilepsy, were treated with combination therapy of topiramate and VNS. All three had a significant reduction in seizures, but even more dramatic improvement in aggression, social interaction, and ambulation. The Cyberonics Patient Outcome Registry was subsequently queried and a beneficial effect of this combination therapy on behavior (specifically alertness) beyond that of VNS and other anticonvulsants was noted. This did not appear to be due solely to seizure reduction, which was observed only differentially at 12 months. PMID- 15123030 TI - Aromatase inhibition, testosterone, and seizures. AB - The effect of testosterone on brain excitability is unclear. The excitatory aspect of testosterone's action in the brain may be due to its conversion to estrogen via aromatase. We report herein a 61-year-old man with temporal lobe epilepsy and sexual dysfunction due to low testosterone levels. Use of an aromatase inhibitor, letrozole, normalized his testosterone level and improved his sexual functioning. Letrozole, in addition to standard antiseizure medication, was also associated with improved seizure control. This was sustained and, further, was associated with seizure exacerbation after withdrawing letrozole, and subsequent seizure improvement after restarting it. During the course of treatment, his serum testosterone level increased, sex hormone-binding globulin decreased (SHBG), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) levels increased, while serum estradiol levels remained undetectable. Letrozole may, therefore, have produced a central alteration in the testosterone/estrogen ratio, thereby impairing estrogen-mediated feedback control of the pituitary, resulting in the observed increase in circulating LH and FSH levels. This experience suggests that aromatase inhibitors should be further investigated as a beneficial treatment modality for male patients with epilepsy. PMID- 15123031 TI - Structural versus functional prediction of memory change following anterior temporal lobectomy. AB - Decline in recent memory function is a significant risk for patients undergoing anterior temporal lobectomy. We report a patient with a febrile seizure history, complex partial seizures arising from the left anterior temporal lobe, and MRI evidence suggesting left hippocampal sclerosis, all of which indicate a low likelihood of significant postoperative memory decline. However, high normal verbal memory on neuropsychological testing and bilaterally normal Wada memory scores indicated increased risk for postoperative memory decline. Following left anterior temporal lobectomy, the patient displayed a significant decline in verbal recent memory that affected school performance. Despite the worsening in memory, the patient reported a significant improvement in his self-reported quality-of-life perception, demonstrating that factors other than change in cognitive performance are related to whether a patient considers epilepsy surgery worthwhile. In the present case, behavioral measures were superior to structural measures in predicting cognitive change following surgery. PMID- 15123033 TI - How is clinical progress achieved? PMID- 15123032 TI - Pseudoseizures and hysterical stridor. AB - Differentiation of pseudoseizures from epileptic seizures and hysterical stridor from asthma is key to the treatment of patients. Both pseudoseizures and hysterical stridor are associated commonly with sexual abuse, eating disorders, depression, substance abuse, anxiety disorders, and personality disorders. Failure to reach an appropriate diagnosis results in inappropriate medical care including recurrent intubations, antiepileptic drugs, and excessive hospitalizations. This case represents the first instance of reported pseudoseizures in a patient with hysterical stridor secondary to childhood sexual abuse. PMID- 15123034 TI - Corticosteroids--from an idea to clinical use. AB - Corticosteroids form the basis of treatment in many inflammatory rheumatic diseases, both as systemic treatment and as treatment with local injections to reduce inflammation. In 1948 the first systemic treatment of a patient with a rheumatic disease was given to a woman with severe rheumatoid arthritis (RA); the impressive effect in this patient, and in another 15 patients, was reported by Dr Hench and co-workers in 1949. Systemic corticosteroid treatment was rapidly adopted and used not only for patients with RA but also for those with other rheumatic diseases such as systemic lupus erythematosus-as well as other disorders such as asthma-with a similar positive effect. In the following year, 1950, the Nobel Prize was awarded for the discovery of the structure and biological effects of the adrenal cortex hormones. This open trial was followed by several controlled trials conducted in the UK in which the effects of cortisone were compared with the effects of aspirin in patients with RA interestingly, without any significant clinical benefit for the cortisone-treated patients. It was not until 1959, in yet another multi-centre trial in Britain, that a significant effect on functional capacity and general well-being was reported after 2 years of treatment with prednisolone, compared to aspirin, in patients with early RA. Despite the dramatic effects that were observed in the severely ill RA patients reported by Hench and co-workers it took 10 years to demonstrate that this effect was superior to the effect of aspirin when the two compounds were compared in controlled trials. Why was this so? One explanation could be in the study designs and the different outcome measures used in the various studies. Perhaps the results in the first comparative studies would have been different if individual response criteria had been used. This is discussed in this chapter. PMID- 15123035 TI - Glucocorticoids in rheumatoid arthritis: a senescent research agenda on the brink of rejuvenation? AB - Treatment of rheumatoid arthritis with glucocorticoids remains controversial despite a considerable and growing body of evidence. This chapter focuses on the research agenda in urgent need of execution: to define conclusively the benefit/harm trade-offs and pin down the place of these agents in the treatment cascade. PMID- 15123036 TI - The process of identifying and understanding cytokines: from basic studies to treating rheumatic diseases. AB - This is a historical overview seen from a personal angle. It covers the insights made during the past 20 years into the destructive processes of rheumatoid arthritis (RA) related to cytokines. The biochemical knowledge of the matrix components (i.e. collagen) and enzymology (i.e. collagenase) available in the 1950s led to the identification of cells from synovial tissue producing collagenase (fibroblast-like cells) and their interaction with other immune cells, i.e. monocyte-macrophages (Mphi) and lymphocytes (1976-1979). This insight led to the isolation of soluble factors produced by Mphi, such as interleukin-1 (IL-1) and TNF, the principal cytokines inducing collagenase and PGE(2) in many target cells (i.e. synovial fibroblasts, chondrocytes, bone-derived cells) (1981 1985). Further advances resulted from observations that, in clinical conditions (i.e. leukaemia, juvenile RA), a remission of fever and inflammation may occur spontaneously and that tissue catabolism may persist despite the absence of systemic inflammation; this gave rise to the concept and identification of endogenous cytokine inhibitors (i.e. IL-1 receptor antagonist and TNF soluble receptor) (1984-1989). The fourth milestone was the observation that the production of IL-1 and TNF by Mphi was induced mainly by direct contact with lymphocytes, prompting studies of the ligands and counter-ligands on Mphi and lymphocytes as well as inhibitors involved in this cell-cell contact, some of these inhibitors being involved in lipid metabolism and acute-phase proteins (HDL apo A-1). PMID- 15123037 TI - The usefulness and the limitations of animal models in identifying targets for therapy in arthritis. AB - Animal models have played a critical role in the history of modern drug development for rheumatoid arthritis (RA). In this chapter I examine the contributions of animal models in arthritis therapy from adjuvant arthritis and COX-1 inhibitors to transgenic mice and biological response modifiers. Advances in knowledge of the mechanisms of connective tissue disease are frequently derived from the study of animal models, and these findings frequently identify therapeutic targets that are subsequently evaluated in animal models. Hence a critical relationship between insights into the pathology of arthritis and the development of novel therapeutic approaches exists around the study of animal models of arthritis. In particular, we examine how the study of collagen-induced arthritis in rodents led to pioneering work in cytokine inhibitors for the successful therapy of RA. PMID- 15123038 TI - The transfer of a laboratory based hypothesis to a clinically useful therapy: the development of anti-TNF therapy of rheumatoid arthritis. AB - The development of anti-TNF therapy is a key step forward in rheumatology as it is the first new therapy for based on investigating the molecular mechanisms of this disease. This chapter reviews how this discovery was made. PMID- 15123039 TI - Use of new biotechnology to design rational drugs against newly defined targets. AB - Success in drug discovery depends largely on the implementation of appropriate strategies that build on new technologies and the appropriate mix of drug discovery platforms and research management procedures. Close collaboration between pharmaceutical companies, biotechnology companies and academic institutions during the many intricate phases of drug discovery is necessary to address the need to co-ordinate and streamline target discovery and validation activities, which typically take much longer than anticipated. Antibodies have become an important segment of newly developed therapeutics for a wide range of indications and offer the appropriate risk/benefit profile to balance drug discovery and development portfolios for optimum success. However, as with other discovery activities, long-term commitment and experience are required to exploit these new techniques fully. Companies with experience in managing the appropriate mix of small-molecule and antibody discovery efforts while implementing novel techniques will remain at the forefront of drug development. PMID- 15123040 TI - Are better endpoints and better design of clinical trials needed? AB - For the clinician it is important to know whether a new drug works, and how the new drug performs against other drugs. However, new drugs are typically tested in placebo-controlled trials without active comparison. New drugs are often tested in a population with high levels of disease activity. Clinicians, however, may also seek the optimal treatment for patients with persistent moderate levels of disease activity. An answer may come from clinical trials that compare two effective drugs in patients with moderate disease activity. A main consequence, however, is that trial endpoints are needed that can detect small but relevant differences in efficacy. An ideal endpoint for trials in rheumatic diseases may be a marker that continually follows short-term changes in the disease process. The levels of the marker should be strongly associated with long-term outcome, thus prognosticating the future. When it can be measured truly and feasible, the marker would be useful as an endpoint in trials and for supporting treatment decisions in clinical practice. PMID- 15123042 TI - Drug-induced and toxic myopathies. AB - Drug-induced myopathies and, more rarely, rhabdomyolysis, are a common biological and clinical setting for clinical rheumatologists. The focus of this chapter is to review (i) the clinical presentation and management of these adverse drug reactions (ADR) according to pain and associated neurological symptoms, (ii) the common drugs prescribed by rheumatologists which may induce reactions such as ADR, with special reference to new drugs, (iii) the pathological classification associated with specific patterns, and (iv) the risk factors leading to myotoxicity (including genetic predisposition). Specific features to be reviewed include macrophage myofasciitis and biological agents of major importance when considering terrorist attacks with biological weapons. When diagnosis is suspected, discontinuation of the putative drug(s) is mandatory and should be carefully monitored. PMID- 15123043 TI - The hereditary amyloidoses. AB - Hereditary amyloidosis is, in general, a systemic condition related to multiple organ system involvement by beta-structured protein deposits. As such, it often mimics the more common forms of systemic amyloidosis: immunoglobulin light chain (AL, primary) and reactive (AA, secondary). The challenge diagnostically is to recognize hereditary amyloidosis as a distinct entity and then to determine the specific type of genetic disease. There are several types of hereditary amyloidosis and precise diagnosis is essential for proper therapy and genetic counselling. This chapter strives to present the subject of hereditary amyloidosis in a way which facilitates understanding of the disease, of the means for diagnosis, of the present and possible future therapies, and of the importance of combined basic and medical research. PMID- 15123044 TI - Musculoskeletal disorders in secondary amyloidosis and hereditary fevers. AB - Amyloidosis remains a severe potential complication of many chronic inflammatory disorders, foremost of rheumatoid arthritis. It is not exactly known why some patients develop a progressive amyloidosis while others do not, although latent deposits may be present. It is likely that more potent anti-inflammatory drugs recently used in rheumatoid arthritis have led to a decrease of amyloid associated (AA) amyloidosis. However, overt amyloidosis remains a severe complication of some chronic inflammatory disorders and it has a poor prognosis. Hereditary fevers are a group of diseases characterized by intermittent bouts of clinical inflammation with focal organ involvement, mainly abdomen, musculoskeletal system and skin. The most frequent is familial Mediterranean fever which affects patients of Mediterranean descent all over the world. Three other types have been recently characterized clinically as well as genetically. A thorough diagnosis is warranted, as clinical and therapeutic management is specific for each of these diseases. PMID- 15123045 TI - Musculoskeletal disorders in diabetes mellitus: an update. AB - Diabetes mellitus (DM) is associated with several musculoskeletal disorders. The incidence of DM and the life expectancy of the diabetic patient have both increased, resulting in the increased prevalence and clinical importance of musculoskeletal alterations in diabetic subjects. The exact pathophysiology of most of these musculoskeletal disorders remains obscure. Connective tissue disorders, neuropathy, vasculopathy or combinations of these problems, may underlie the increased incidence of musculoskeletal disorders in DM. The development of musculoskeletal disorders is dependent on age and on the duration of DM; however, it has been difficult to show a direct correlation with the metabolic control of DM. Most of these disorders can be diagnosed clinically, but some radiological examination may help, especially in differential diagnosis. No specific treatment is available, and treatments used in the general population are also recommended for diabetic subjects. Infectious complications affecting the musculoskeletal system are common in DM subjects, and these, possibly life threatening, complications should be systematically discussed. PMID- 15123046 TI - Musculoskeletal and other extrapulmonary disorders in sarcoidosis. AB - Sarcoidosis is a multisystemic inflammatory disease, still of unknown origin, characterized by epithelioid non-caseating granuloma in all affected organs. Granuloma formation is lead by a Th1-type response. The exact mechanism that leads to either progression or spontaneous resolution of the disease is not known. Familial aggregation, and the variations in presentation and severity of sarcoidosis according to ethnic background, are suggestive of a polygenic origin that is still to be determined. The contribution of environmental factors, as well as their interactions with genetic factors, remains to be demonstrated. The clinical presentation, musculoskeletal and other extrapulmonary disorders, as well as patients work-up, are reviewed. Sarcoidosis is often a benign disease. Corticosteroids, either alone or in association with other drugs, are still the mainstay of treatment. Defining clearly who will need treatment and what treatment to be used in a particular patient remains controversial. PMID- 15123047 TI - Joint hypermobility. AB - Joint hypermobility is an area of neglect in rheumatology. That is not to say it is overlooked by rheumatologists. It is spotted when sought, but for many unfortunate patients, here the story ends. The act of recognition becomes the goal in itself rather than the medium through which effective therapy can be provided. This chapter serves to reinforce the clinical and epidemiological importance of a common disorder whose significance is under-appreciated and impact largely ignored. In contradistinction to our earlier chapter, published in 2000, which took for its remit the heritable disorders of connective tissue in general, the current one focuses on the commonly encountered (so-called benign) joint hypermobility syndrome, its recognition, epidemiology, clinical features and management according to the most recent literature. PMID- 15123048 TI - Skeletal dysplasias and the osteoarthritic phenotype. AB - In contrast to late-onset osteoarthritis (OA), the appearance of precocious OA has historically been recognized as a particularly aggressive form of the disorder that is frequently inherited as a Mendelian trait. In general, precocious OA appears as a consequence of many skeletal dysplasias, which, although individually rare, comprise a sizable population of patients when viewed in toto. In these patients the disease is often rapidly progressive and includes features of articular and extra-articular involvement that are not typical of classic OA. The molecular pathology of the chondro-osseous disorders has been the focus of intense study in recent years, with the promise of providing insight into skeletal development and homeostasis, as well as the aetiology and pathogenesis of degenerative joint disease. PMID- 15123049 TI - Paget's disease of bone. AB - Paget's disease of bone is characterized by an anarchic bone remodelling, associated with morphological and functional abnormalities of osteoclasts. Its prevalence and incidence rates decreased gradually over the past two decades; the reason for this remains unclear. The aetiology of the disease is still obscure, the paramyxoviral theory being very controversial. Recent advances in understanding of the disease come from genetic studies, with the identification of specific mutations in the p62-sequestosome gene, which could be involved in pathogenetic mechanisms leading to increased osteoclast activity. The disease affects one or several bone pieces, leading to bone pain, deformities, characteristic imaging features, and increased markers of bone remodelling. The long-lasting disease activity leads to complications, including arthropathies, neurological compressions, fissures or fractures and, rarely, osteosarcomatous transformation of a pagetic lesion. Potent bisphosphonates have proven their efficacy in reducing symptoms and disease activity. They are currently used as the first-line treatment with the goal of normalizing bone remodelling and, hopefully, preventing late complications. PMID- 15123050 TI - Stress fractures. AB - Stress fractures are common over-use injuries which include fatigue and insufficiency fractures. Athletes, soldiers and osteoporotic patients are some of the individuals at high risk for the development of this injury. Owing to the low sensitivity of plain radiography at the onset of symptoms the diagnosis of this entity may be easily overlooked. Occasionally, some of these fractures, such as tibial fracture in children and fractures in the clavicle and pelvic ring, can be misdiagnosed as tumoral or infectious processes; moreover, although most stress fractures are uncomplicated and can be managed by rest and restriction from precipitating activity, a subset of these fractures can present a high risk for progression to complete fracture or non-union problems. All of this indicates that the various types of stress fracture, owing to their different clinical characteristics and evolution, should be commented on in detail. In this chapter different types of stress fracture are described with special reference to their localization, clinical characteristics, evolution and treatment. PMID- 15123052 TI - Preface. PMID- 15123051 TI - Musculoskeletal manifestations of osteomalacia and rickets. AB - Osteomalacia (OM) is still an important metabolic bone disease with increased prevalence in certain regions of the world as well as in the urban population of elderly confined. The disease presents with a wide variety of clinical, biochemical and radiographic manifestations mimicking other musculoskeletal disorders, including 'osteoporosis'. In this chapter, we provide the basis for its clinical diagnosis and management. There have been significant recent advances in the understanding of vitamin D deficiency and hypophosphataemic osteomalacia, which can now assist clinicians in the precise diagnosis and treatment of this disease. In this chapter we also review the various underlying aetiologies. The successful management of OM depends on the underlying aetiology. PMID- 15123054 TI - Irregular or absent periods--what can an ultrasound scan tell you? AB - Transvaginal ultrasonography has increased our appreciation of the physiological changes in the ovary and endometrium that occur during the normal menstrual cycle. It has become a primary investigative tool in women with irregular or absent periods. Its usefulness in cases of primary amenorrhoea to assess anatomy is also undisputed although it may have limitations in terms of its specificity. However, the interpretation of ultrasound images in women with irregular menses or secondary amenorrhoea is not entirely straightforward. This is particularly true in the diagnosis of polycystic ovary syndrome, a condition of uncertain aetiology, which may present with oligoamenorrhoea. This chapter aims to discuss the benefits and limitations of ultrasound while taking into account the broad overlap between normal and abnormal physiology, some of which has still to be elucidated. PMID- 15123055 TI - An ultrasound-based approach to the assessment of infertility, including the evaluation of tubal patency. AB - An optimal initial infertility investigation protocol would be a process that is diagnostically accurate, expeditious, cost-effective, reliable and as minimally invasive as possible. In addition, the investigation should provide the clinician with useful prognostic information regarding possible future treatment. At present, extensive use of invasive procedures such as diagnostic hysteroscopy and laparoscopy is the standard at many fertility centres. Recent advances in gynaecological ultrasonography have shown that ultrasound can replace routine invasive investigative procedures. An ultrasound-based approach would make the basic infertility investigation less time-consuming and less expensive, but at the same time more acceptable to the majority of patients. This chapter describes an ultrasound-based approach to the assessment of infertility. In addition, the role of ultrasonography for assessment of the pelvic organs as a basic part of the initial investigation of an infertile couple is discussed and compared to more traditional invasive methods. PMID- 15123056 TI - Assessing congenital uterine anomalies: the role of three-dimensional ultrasonography. AB - Congenital uterine anomalies are associated with a range of adverse reproductive outcomes but may also be found incidentally. Their significance in women with normal reproductive histories, who do not have a history of recurrent miscarriage or infertility, has been uncertain. The absence of these data has made it difficult to understand the true significance of congenital uterine anomalies found in women with adverse pregnancy outcomes. The advent of transvaginal three dimensional ultrasonography has enabled the accurate, non-invasive, outpatient diagnosis of congenital uterine anomalies. It has enabled large-scale screening and morphological analysis of congenital uterine anomalies. PMID- 15123057 TI - The management of early pregnancy complications. AB - Early pregnancy complications include miscarriage, ectopic pregnancies, pregnancies of unknown location, adnexal masses and hydatidiform mole. The management of women with these complications has changed with the introduction of dedicated Early Pregnancy Units, with a shift away from a surgical approach to one based on an expectant or 'watch and wait' policy. This chapter describes the role of transvaginal sonography (TVS) in the management of these women and critically evaluates the different treatment modalities available for the management of these complications. The management of miscarriage has largely changed from a surgical to an expectant or medical approach. The treatment of ectopic pregnancy (EP) is also being managed in a similar way as the condition is increasingly detected earlier. We will argue that EP should not be diagnosed on the basis of the absence of an intrauterine pregnancy but rather by the positive visualization of an adnexal mass using TVS. If a pregnancy cannot be seen either inside or outside the uterus this should be described as a pregnancy of unknown location and managed expectantly until the outcome is confirmed. PMID- 15123058 TI - Menorrhagia and abnormal bleeding before the menopause. AB - Abnormal uterine bleeding before the menopause accounts for 20% of visits to the gynaecology clinic and almost 25% of gynaecological operations. The patient who presents with such bleeding presents two distinct but important challenges for the clinician. The first is the exclusion of cancer or hyperplasia; the second is dealing with the annoyance as well as the fear that the bleeding engenders in the patient. Any attempt at appropriate therapy--whether surgical, hormonal or expectant--begins with an accurate diagnosis. Transvaginal ultrasound with saline infusion sonohysterography for selective patients has emerged as a safe, non invasive and, inexpensive method of triaging patients with abnormal uterine bleeding in order to determine which patients require no further evaluation, blind endometrial sampling for a global endometrial process or visually directed endometrial sampling when pathology is thought to be focal. PMID- 15123059 TI - Use of morphology to characterize and manage common adnexal masses. AB - The use of grey-scale ultrasound morphology to characterize a pelvic mass may also be called 'pattern recognition'. The grey-scale ultrasound image provides us with the same information as that obtained by the surgeon or pathologist when he or she cuts a surgical specimen to see what it looks like inside. Many pelvic masses have such a typical macroscopic appearance that a fairly confident diagnosis can be made on the basis of their macroscopic appearance alone, i.e. on the basis of their grey-scale ultrasound image. This is true of most dermoid cysts, endometriomas, corpus luteum cysts, hydrosalpinges and peritoneal pseudocysts, and of many paraovarian cysts and benign solid ovarian tumours, for example, fibromas, fibrothecomas, thecofibromas, thecomas and Brenner tumours. A mass with irregularities should always evoke suspicion of malignancy. A mass that is completely smooth is almost certainly benign. Papillary projections- considered a strong sign of malignancy--are more common in borderline tumours than in invasive cancers but may also be seen in benign tumours, for example, in adenofibromas. They explain many false-positive ultrasound diagnoses of malignancy. Pattern recognition is superior to all other ultrasound methods (e.g. simple classification systems, scoring systems, mathematical models for calculating the risk of malignancy) for discrimination between benign and malignant extrauterine pelvic masses. Today's often too liberal use of transvaginal ultrasound gives clinicians problems. Many adnexal masses that probably would have remained undetected before the ultrasound era are now found incidentally at transvaginal ultrasound examination in women without symptoms of an adnexal tumour. The natural history of incidentally detected pelvic masses with benign ultrasound morphology is not known. Therefore, the optimal management of such tumours is also unknown. PMID- 15123060 TI - The use of mathematical models to evaluate pelvic masses; can they beat an expert operator? AB - The pre-operative characterization of ovarian cysts remains a major challenge. Functional cysts and some other benign cysts should be managed conservatively, whereas persistent tumours may need removal. It is crucial to distinguish between malignant tumours, which are better treated by a gynaecological oncologist, and benign tumours, which may be suitable for minimal-access surgery. Over the past decade several ultrasound-based morphological scoring systems, colour Doppler parameters, logistic regression models and artificial neural networks have been proposed and tested in order to try to predict the histology of ovarian tumours. On prospective testing none of the current models can beat an expert sonologist. Signs of malignancy include the presence of papillary structures, irregular solid areas, septa and a strong vascularization at colour Doppler imaging. Further refinement of mathematical models and the results of multicentre trials need to be reviewed before the clinical use of mathematical models can be advocated. PMID- 15123061 TI - The role of ultrasound in the management of women with acute and chronic pelvic pain. AB - Pelvic pain (acute or chronic) is a common symptom in women of all ages. Ultrasonography is the least invasive investigative tool available to the clinician. Transvaginal probes produce high-resolution images of the pelvic organs, providing reliable and reproducible information without the need for a full bladder. Common gynaecological pathology involving the uterus, Fallopian tube and/or the ovary can be diagnosed with confidence. Non-gynaecological pathology involving the bowel can also be diagnosed with accuracy, and will often be seen in the acute gynaecological setting. Ultrasound can be used to triage patients into appropriate treatment protocols, enabling the clinician to avoid surgery in some cases and select the correct surgical approach in others. If the patient has a negative pregnancy test, no pelvic tenderness on bimanual examination and a normal scan, significant pathology is very unlikely. PMID- 15123062 TI - Managing women with post-menopausal bleeding. AB - Transvaginal ultrasound examination can reliably distinguish women with post menopausal bleeding (PMB) who are at low risk of endometrial pathology (endometrial thickness < or =4 mm) from those who are at high risk (endometrium > or =5 mm) and can rule out focally growing lesions in the uterine cavity using saline infusion into the cavity as a negative contrast agent (hydrosonography). The 5 mm cut-off is applicable irrespective of the use of hormone replacement therapy. It is justified to refrain from endometrial sampling in women with PMB and an endometrial thickness of < or =4 mm because the risk of endometrial cancer in these women is low (0.1-1.0%). However, it is not known whether these women need follow-up. About 80% of women with PMB and an endometrium of > or =5 mm have focally growing pathological lesions in the uterine cavity. These should be removed by operative hysteroscopy because dilatation and curettage (D and C) will fail to diagnose and remove a large proportion of these lesions. However, D and C is a reliable diagnostic method for women without focal lesions in the uterine cavity. It is not known whether simple outpatient sampling devices (e.g. Pipelle) are as reliable as D and C in women without focal lesions. A measurement of endometrial thickness is a simple and accurate method for estimating the risk of endometrial cancer. The reliability of ultrasound evaluation of endometrial morphology and/or vascularization for risk estimation of endometrial malignancy remains to be determined. PMID- 15123063 TI - Investigating abnormal bleeding on HRT or tamoxifen: the role of ultrasonography. AB - In western countries hormone replacement therapy (HRT) is a widely used therapeutic and/or preventive medication in post-menopausal women. Its impact on the endometrium has been extensively investigated. The critical issue is whether and how to monitor the endometrium in women who are taking HRT. Transvaginal ultrasound is a potential candidate for this role. Evidence for the efficiency of transvaginal sonography (TVS) in this context is limited. The criteria used to define a normal or abnormal result in women taking HRT are based largely on data from post-menopausal women who are not taking this treatment. This extrapolation is probably reasonable as long as some degree of caution is used. The use of normative data from post-menopausal women for those taking selective oestrogen receptor modulators (SORMs) is not advised and should be utilized only in the context of research studies as insufficient data exist on which to base cut-off values to discriminate between normal and abnormal endometria. PMID- 15123064 TI - Anal endosonography and its role in assessing the incontinent patient. AB - Anal endosonography is now recognized as an important investigation in the assessment of faecal incontinence. The endosonographer needs to be aware that the anatomy of the anal sphincter is complex and therefore there can be pitfalls in the interpretation of images. The findings have clinical implications on subsequent management and can contribute to prognosticating outcome. However, anal endosonography has a complementary role and other investigations, such as anal manometry, should be performed before intervention. PMID- 15123066 TI - Bugs and the gut: an unstable marriage. AB - There is a symbiotic relationship between the gastrointestinal microflora and the human host. Commensal bacteria provide essential nutrients to the epithelium and promote healthy immune responses in the gut. Commensal bacteria such as Escherichia coli can, however, transform into pathogens when they acquire genetic material encoding virulence factors such as adhesins, enterotoxins, invasins and cytotoxins. Enterovirulent organisms 'communicate' with the host by a variety of diverse mechanisms; these underpin the pathogenic processes that are essential for the expression of diarrhoeal disease. Many of these mechanisms involve the activation of signal transduction pathways in epithelial cells. Classical pathways include activation of adenylate or guanylate cyclases to produce chloride secretion, and subversion of cytoskeletal functions to effect intimate attachment with or without invasion of epithelial cells. Other systems are also involved, including inflammatory cells and local neuroendocrine networks. Understanding the complex interactions between the human gastrointestinal tract and the commensals and pathogens which it encounters will hopefully help us to exploit further the beneficial effects of the 'marriage' and to find new ways of preventing and treating microbial disease of the intestine which occurs when the symbiotic arrangement breaks down. PMID- 15123067 TI - Microbial-gut interactions in health and disease. Epithelial cell responses. AB - Intestinal epithelial cells are unique in that they represent the only host cells that are constantly interacting with a very large bacterial population in the lumen. The single monolayer of epithelial cells consists of subpopulations with distinct functions that include protection against luminal microorganisms. Although the microbial flora remains to be fully characterized, its normal relationship with the host intestinal epithelial cells appears to be predominantly symbiotic or commensal. The molecular complexity of the epithelial microbial relationship has been shown in studies that have examined the establishment of the resident bacteria in germ-free mice. Recent work has also demonstrated the ability of resident bacteria to enhance epithelial protective responses. The mechanisms by which epithelial cells may avoid pro-inflammatory responses to resident microorganisms, while retaining the capacity to respond to pathogens, are also being characterized. PMID- 15123068 TI - Microbial-gut interactions in health and disease. Interactions between dendritic cells and bacteria in the regulation of intestinal immunity. AB - Dendritic cells (DCs) are immunoregulatory antigen-presenting cells. DCs can be potent activators of naive T cells and influence the generation and homing of effector lymphocytes; they can also induce regulatory mechanisms and maintain non responsiveness. In part, these different outcomes are influenced by exposure of the DC to microbial products. The regulatory role of DCs is of particular importance at mucosal surfaces such as the intestine, where the immune system exists in intimate association with the external antigenic environment. Much of what we know about DCs has come from studies on the cells outside the gastrointestinal tract but information about gut DCs and their contribution to the specialized immune environment of the gut is now emerging. Here, we review current knowledge on gut DCs, suggest models for interactions between DCs and the commensal microflora in health and disease, and discuss gut DCs as targets for probiotic therapies. PMID- 15123069 TI - Microbial-gut interactions in health and disease. Immune responses. AB - The indigenous bacterial microflora colonize the gut at birth and remain there throughout life. Approximately 10(14) bacteria are present in the ileum and colon and they are clearly immunogenic. The evidence is strong that the vast majority of IgA plasma cells in normal human gut are responding to the antigens of the flora, and although the flora is also responsible for producing the large numbers of T cells which are present in the gut of healthy individuals, the types of T cell response which the flora elicits are less well understood. A major challenge for the immune system is to distinguish between the antigens of the flora and the antigens of pathogens. There is also a growing realization that the normal flora can also influence gene expression in antigen-presenting cells in the gut and so set the context in which T cells respond to food antigen and vaccines. PMID- 15123070 TI - Microbial-gut interactions in health and disease. Prebiotics. AB - In nutritional sciences there is much interest in dietary modulation of the human gut. The gastrointestinal tract, particularly the colon, is very heavily populated with bacteria. Most bacteria are benign; however, certain gut species are pathogenic and may be involved in the onset of acute and chronic disorders. Bifidobacteria and lactobacilli are thought to be beneficial and are common targets for dietary intervention. Prebiotic is a non-viable food ingredient selectively metabolized by beneficial intestinal bacteria. Dietary modulation of the gut microflora by prebiotics is designed to improve health by stimulating numbers and/or activities of the bifidobacteria and lactobacilli. Having an 'optimal' gut microflora can increase resistance to pathogenic bacteria, lower blood ammonia, increase stimulation of the immune response and reduce the risk of cancer. This chapter examines how prebiotics are being applied to the improvement of human health and reviews the scientific evidence behind their use. PMID- 15123071 TI - Microbial-gut interactions in health and disease. Probiotics. AB - The definition of probiotics has evolved from a live active culture which improves the balance of the gut microbiota composition to specific effects, in particular, the immunomodulatory potential of clearly defined strains. The strains with beneficial properties, potential sources of probiotics, most frequently belong to the genera Bifidobacterium and Lactobacillus, and some of these strains exhibit powerful anti-inflammatory properties. Indeed, probiotic therapy has attracted research interest in human infectious, inflammatory and allergic disease. The most fully documented disease altering the gut microbiota is acute infectious diarrhoea in childhood. Current probiotic research aims to provide safe but sufficient bacterial stimulus in order to avert deviant immune responsiveness related to allergic and inflammatory diseases. However, further rigorous scientific efforts are required to characterize the immunomodulatory potential of specific probiotic strains for these targets. PMID- 15123072 TI - Microbial-gut interactions in health and disease. Irritable bowel syndrome. AB - The intestinal microbiota interacts with several aspects of gastrointestinal function that may affect the expression or progression of disease. For example, a role for bacterial metabolism of bile acids and food has been linked to colorectal cancer development. Studies have also shown a potential role of the intestinal microbiota in the modulation of inflammation in the intestine and joints. Normal gut physiology is molded by the interaction between the intestinal microbiota and the host's gastrointestinal tissues, including motility, absorption and secretion, and intestinal permeability. Early studies in axenic mice demonstrated gross morphological abnormalities and gut motor dysfunction related to the absence of a normal microflora, raising the possibility that shifts in commensal bacterial populations could play a role in the development of altered motility states including functional disorders of the gut. This chapter concentrates on the experimental evidence for a role of intestinal microbiota and the potential therapeutic value of probiotics in functional diseases such as irritable bowel syndrome. PMID- 15123073 TI - Microbial-gut interactions in health and disease. Gastrointestinal cancer. AB - A combination of both environmental and genetic factors contributes to the vast majority of human cancers and in particular cancers of the gastrointestinal tract, including the stomach, colon and rectum. The mechanisms associated with cancer causation or prevention are largely unknown and the subject of much research. Many of these mechanisms implicate the metabolic activities of the bacterial flora normally resident in the gastrointestinal tract. This paper examines both the detrimental and beneficial consequences of bacterial activity of the gastrointestinal tract, focusing in particular on the stomach and large intestine. PMID- 15123074 TI - Microbial-gut interactions in health and disease. Antibiotic-associated diarrhoea. AB - Most cases of antibiotic-associated diarrhoea (AAD) are directly or indirectly due to the alteration of gut microflora by antibiotics. 'Functional' diarrhoea, usually limited to a mild and brief change in stool frequency, is considered as the most frequent pattern of AAD. Reduced carbohydrate fermentation and impaired metabolism of bile acids have been claimed as the potential causes of this transient digestive discomfort but a critical analysis of the data supporting these theories is necessary. Alternatively, changes in the gut flora ecosystem allow pathogens to proliferate. Clostridium difficile is responsible for approximately 10% of cases of AAD and almost all cases of antibiotic-associated pseudomembranous colitis. The level of evidence which supports the potential responsibility of other candidate pathogens (Klebsiella oxytoca, enterotoxin producing Clostridium perfringens and Staphylococcus aureus, Candida) needs to be appreciated according to the updated postulates of causality relationships between a bacterium and a disease. PMID- 15123075 TI - Gut microflora in the pathogenesis of the complications of cirrhosis. AB - The gut flora plays an important role in the pathogenesis of the complications of cirrhosis. Cirrhotic patients are prone to develop bacterial infections, mainly the 'spontaneous' infection of ascites or spontaneous bacterial peritonitis. Other complications of cirrhosis, such as variceal haemorrhage and ascites, occur mostly or solely as a consequence of portal hypertension. Portal pressure increases initially as a consequence of an increased intrahepatic resistance but, once collaterals have formed, high portal pressure is maintained by an increased splanchnic blood inflow secondary to vasodilatation. Splanchnic vasodilatation is the initiating event in the hyperdynamic circulatory state that aggravates the complications of cirrhosis. The gut flora plays a role in both the development of infections and in the hyperdynamic circulatory state of cirrhosis and, although less prominently, it also plays a role in the pathogenesis of hepatic encephalopathy. This chapter presents evidence regarding gut flora and its modification in the pathogenesis and management of these complications of cirrhosis. PMID- 15123076 TI - Microbial-gut interactions in health and disease. Epithelial cell responses. AB - Pathogenic bacteria use many strategies to secure their survival within the host. Enteropathogens exploit intestinal epithelial cells in many ways, including the manipulation of normal cellular functioning, or of cellular structural components, or by the induction of signalling pathways, such as the production of pro-inflammatory cytokines. However, the enterocyte warns the host of impending danger and, in turn, elicits a protective response. Pathogens are detected by epithelial cells owing to their vast array of surface antigens and secreted products. Epithelial cells have developed both extracellular and intracellular sensing proteins that function as a first line of defence against pathogens; this is followed by acquired immunity, namely IgA, which is used as reinforcement. Thus, in a game of constant attack and defence, the pathogen and the enterocyte aim to outsmart each other in an effort to survive. PMID- 15123077 TI - Microbial-gut interactions in health and disease. Mucosal immune responses. AB - The host gastrointestinal tract is exposed to countless numbers of foreign antigens and has embedded a unique and complex network of immunological and non immunological mechanisms, often termed the gastrointestinal 'mucosal barrier', to protect the host from potentially harmful pathogens while at the same time 'tolerating' other resident microbes to allow absorption and utilization of nutrients. Of the many important roles of this barrier, it is the distinct responsibility of the mucosal immune system to sample and discriminate between harmful and beneficial antigens and to prevent entry of food-borne pathogens through the gastrointestinal (GI) tract. This system comprises an immunological network termed the gut-associated lymphoid tissue (GALT) that consists of unique arrangements of B cells, T cells and phagocytes which sample luminal antigens through specialized epithelia termed the follicle associated epithelia (FAE) and orchestrate co-ordinated molecular responses between immune cells and other components of the mucosal barrier. Certain pathogens have developed ways to bypass and/or withstand defence by the mucosal immune system to establish disease in the host. Some 'opportunistic' pathogens (such as Clostridium difficile) take advantage of host or other factors (diet, stress, antibiotic use) which may alter or weaken the response of the immune system. Other pathogens have developed mechanisms for invading gastrointestinal epithelium and evading phagocytosis/destruction by immune system defences. Once cellular invasion occurs, host responses are activated to limit local mucosal damage and repel the foreign influence. Some pathogens (Shigella spp, parasites and viruses) primarily establish localized disease while others (Salmonella, Yersinia, Listeria) use the lymphatic system to enter organs or the bloodstream and cause more systemic illness. In some cases, pathogens (Helicobacter pylori and Salmonella typhi) colonize the GI tract or associated lymphoid structures for extended periods of time and these persistent pathogens may also be potential triggers for other chronic or inflammatory diseases, including inflammatory bowel disease and malignancies. The ability of certain pathogens to avoid or withstand the host's immune assault and/or utilize these host responses to their own advantage (i.e. enhance further colonization) will dictate the pathogen's success in promoting illness and furthering its own survival. PMID- 15123078 TI - Defensin-mediated innate immunity in the small intestine. AB - Epithelial cells contribute to innate immunity by releasing antimicrobial peptides (AMPs) onto mucosal surfaces. In the small bowel, Paneth cells at the base of the crypts of Lieberkuhn secrete alpha-defensins and additional AMPs at high levels in response to cholinergic stimulation and when exposed to bacterial antigens. The release of Paneth cell products into the crypt lumen is inferred to protect mitotically active crypt cells that renew the epithelial cell monolayer from colonization by potentially pathogenic microbes and to confer protection from enteric infection. The most compelling evidence for a Paneth cell role in enteric resistance to infection is evident from studies of mice transgenic for a human Paneth cell alpha-defensin, HD-5, which are completely immune to infection and systemic disease from orally administered Salmonella typhimurium. Alpha defensins in Paneth cell secretions may also interact with bacteria in the intestinal lumen above the crypt-villus boundary and influence the composition of the enteric microbial flora, but that remains to be demonstrated. PMID- 15123079 TI - Microbial-gut interactions in health and disease. Progress in enteric vaccine development. AB - Enteric infections resulting in diarrhoea are among the most important causes of morbidity and mortality, particularly in children in developing countries. They are also a common cause of disease among travellers to Africa, Asia and Latin America. Recently, effective, live and inactivated oral and parenteral vaccines against some of the most severe enteric infections-cholera and typhoid fever-have been licensed in several countries. Different candidate vaccines against rotavirus, Shigella and ETEC diarrhoea have also been developed and tested for safety and immunogenicity in developed as well as in developing countries. The protective efficacy of several of these vaccines has also been tested, either in human volunteer challenge studies or in field trials. In this chapter we describe the properties and availability of the recently licensed vaccines and present an update on the diverse efforts being made to achieve new or improved vaccines against the most prevalent enteropathogens. PMID- 15123081 TI - Oesophageal diverticula. AB - Oesophageal diverticula are rare. They are most commonly seen at the pharyngo oesophageal junction (Zenker's diverticula) or at the distal oesophagus (epiphrenic diverticula). In both cases they are caused by altered motility which results in abnormal intraluminal pressure and the pushing of the oesophageal mucosa through focal weaknesses of the muscular wall (pulsion diverticula). The established surgical treatment for these diverticula therefore consists of eliminating the functional obstruction causing the disease (myotomy), associated with resection of the diverticulum (diverticulectomy) or its suspension (diverticulopexy). Recently, the spread of minimally invasive surgery has also led the application of such techniques to the treatment of oesophageal diverticula. Endoscopic diverticulostomy with stapler, laser or coagulation, through a rigid or flexible endoscope, has been demonstrated to be a valid treatment for Zenker's diverticula-as an alternative to surgery-especially in high-risk patients. On the other hand, laparoscopic treatment of epiphrenic diverticula has recently been introduced with encouraging results. However, because the disease is rare, more experience is required in order to allow definitive conclusions. PMID- 15123082 TI - Laparoscopic treatment of gastro-oesophageal reflux disease. AB - Laparoscopic antireflux surgery is now well established as a treatment of moderate to severe gastro-oesophageal reflux disease. It is indicated for patients with reflux symptoms who have not responded fully to medical therapy or who do not wish to continue medication for the rest of their lives. The evidence base for the determination of appropriate practice has expanded considerably in recent years with the publication of several important randomized trials. These trials have confirmed the superiority of fundoplication compared to medical therapy for the treatment of these patients. They have also demonstrated that the laparoscopic approach achieves an improved short-term outcome compared to the equivalent open approach. Additional trials suggest that the routine application of partial fundoplication procedures achieves equivalent reflux control and fewer side-effects than total fundoplication. Longer-term outcome studies have also been reported recently, with success rates of approximately 90% claimed at 5-8 years. Hence, laparoscopic fundoplication is now the 'gold standard' for the management of patients with more severe gastro-oesophageal reflux disease. New endoscopic treatments for reflux will need to achieve similar outcomes before they can replace the laparoscopic approach. PMID- 15123083 TI - Endoscopic suturing. AB - We describe the development of endoscopic sewing machines and ancillary equipment for knot tying and thread cutting. We outline the experimental studies in dogs, pigs and baboons prior to the first studies in man. We consider the early results achieved by groups in Europe and the U.S.A., and present the available evidence from peer-reviewed studies and data from numerous abstracts on the use of endoscopic suturing in man for treating gastro-oesophageal reflux. We consider the limitations of the available studies, and outline the requirements for improvements in flexible endoscopic suturing methods. PMID- 15123084 TI - Enteryx. AB - Enteryx (ethylene vinyl alcohol copolymer) was developed as a bulking agent to be injected endoscopically at the lower oesophageal sphincter (LOS) to increase the competency of the gastro-oesophageal barrier in patients suffering from gastro oesophageal reflux disease (GORD). Preliminary clinical studies have shown that Enteryx implantation is a fast, minimally invasive and safe procedure. In prospective multicentre studies, significant improvement in reflux symptoms, reduction in the use of proton pump inhibitors (PPIs), and objective improvement in acid oesophageal exposure time were observed after 6 months of follow-up. Improvement of GORD symptoms seems to be correlated with the persistence of the implant. Preliminary data suggest a lengthening and an increase in the LOS relaxation pressure as mechanisms of action of this injection technique. Longer follow-up and controlled sham studies are needed to confirm the efficacy of this technique before it can be proposed as a routine alternative to medical or surgical therapies for GORD. PMID- 15123085 TI - Management of pre-malignant and malignant lesions by endoscopic resection. AB - Endoscopic resection (ER) has gained more and more importance in the treatment of early gastrointestinal neoplasia over the last few years. The choice of the different available techniques depends on the site, the macroscopic type of the tumour and the personal experience of the endoscopist. The 'suck-and-cut' technique with ligation device or cap should be favoured to normal strip biopsy in the oesophagus because of the size of the resected specimen and its technical feasibility. A recently described method of ER in the stomach is the circumferential mucosal incision with a type of needle-knife and subsequent en bloc resection following prior injection under the lesions. ER of high-grade intraepithelial neoplasia and mucosal adenocarcinoma in Barrett's oesophagus should be considered as the treatment of choice. First mid-term results of endoscopic therapy of early squamous-cell neoplasia in the oesophagus show promising results; however, long-term results are awaited. Studies with large numbers of patients in Japan proved the efficiency and safety of ER in low-risk early gastric carcinoma. Duodenal lesions and adenomas of the major duodenal papilla were also proved to be treated successfully by ER. In the colon, ER is used successfully for resection of adenomas and small well-differentiated or moderately differentiated carcinomas that are restricted to the mucosa. ER of gastrointestinal lesions is a safe and effective method but should be performed only by experienced endoscopists. PMID- 15123086 TI - Endoscopic therapy for upper gastrointestinal bleeding. AB - Upper gastrointestinal bleeding (UGIB) is one of the most common medical emergencies and remains a major cause of morbidity and mortality among patients. Although initially employed diagnostically, endoscopy has steadily replaced surgery as a first-line treatment in all but the haemodynamically unstable patient. A vast selection of techniques and devices are now available to the dedicated therapeutic endoscopist, including injection therapy, electrical or thermal coagulation and mechanical banding or clipping. The use of endoscopic ultrasound for targeting treatment is increasing and the development of new technologies, such as capsule endoscopy, is likely to play an important role in future protocols. However, despite numerous randomized controlled trials and meta analyses comparing the efficacy of different endoscopic interventions, the implementation of obtained results into treatment regimes has so far failed to impact significantly on overall UGIB mortality, which remains stubbornly at 10 14%. Reducing this continues to be one of the main challenges facing the therapeutic endoscopist. PMID- 15123087 TI - TIPS: an update. AB - During the last 15 years the transjugular intrahepatic portosystemic shunt (TIPS) procedure has become a safe and effective treatment of portal hypertension. Its major obstacle, the high rate of shunt insufficiency, is going to be solved by the availability of covered stents showing a patency rate of up to 90%. The treatment of acute oesophageal and gastric variceal bleeding is an unsolved problem because variceal bleeding remains the major cause of death in patients with cirrhosis. TIPS has become the rescue treatment of choice because it combines high efficacy with low invasiveness. In this context, the timing of the rescue TIPS is of major importance for achieving definitive haemostasis before multi-organ failure develops. In the prevention of re-bleeding, TIPS is accepted as a second-line treatment, required in about 10-20% of patients. TIPS may be indicated when more than two significant re-bleedings occurred within a time frame of 12 months in spite of adequate first-line measures i.e. drugs or ligation. Refractory ascites is the third main indication for TIPS. Five randomized studies comparing TIPS with paracentesis show good response and comparable survival. Interpretations of authors and comments of reviewers are, however, controversial and do not permit a definitive recommendation. PMID- 15123088 TI - Percutaneous ablation of liver tumours. AB - The goal of local ablation treatment of hepatic disease is to prolong survival for patients with unresectable tumours. Presently, influence on survival is difficult to estimate because of the heterogeneity of indications and treatments and short follow-up. This chapter therefore focuses on potential benefits and limitations, complications and solutions for improvement. The main problems with in situ ablation are the lack of good imaging techniques to determine the extent of disease and the lack of a method for real-time monitoring of irreversible tissue effect. With one exception, there are no prospective, randomized studies comparing local destruction methods. It appears that percutaneous ethanol injection and cryotherapy should be replaced by radiofrequency ablation (RFA) or interstitial laser thermotherapy (ILT) and that there is little difference in outcome between RFA and ILT. Intraoperative RFA or ILT is valuable as an adjunct to hepatic resection in order to increase the rate of resectability. The percutaneous approach needs further development. It might be valuable in a few truly unresectable or inoperable patients or in selected patients with neuroendocrine liver metastases. In the large majority of unresectable patients it should, however, presently be used and evaluated only in prospective, randomized studies. PMID- 15123089 TI - Photodynamic therapy in cholangiocarcinomas. AB - Symptoms occur late in cholangiocarcinoma and therefore only about half of the patients at the time of diagnosis are candidates for curative surgery. In patients with advanced non-resectable cholangiocarcinoma palliative treatment options are limited. Until now, insertion of endoprostheses for the treatment of cholestasis has been the method of choice. However, tumour growth cannot be influenced and so that prognosis is dismal. Although radiotherapy and chemotherapy are frequently used, prospective, randomized trials showing an improvement in survival time are missing. Encouraging results from prospective, single-arm phase II trials and a randomized trial using photodynamic therapy (PDT) in non-resectable cholangiocarcinoma indicate considerable benefit on survival with a good quality of life. Furthermore, PDT is well tolerated, with only few specific side-effects. This is of great importance in patients with short life expectancy. PDT should therefore be offered to all patients with non resectable cholangiocarcinoma. However, before initiating PDT or any other palliative measure, a proper staging and a surgical consultation is necessary to avoid missing a curative surgical option. PMID- 15123090 TI - Endoscopic technique for the management of pancreatitis and its complications. AB - Therapeutic endoscopy is now increasingly used to treat gallstone pancreatitis, acute pancreatitis of other aetiologies, chronic pancreatitis and complications associated with acute or chronic pancreatitis. This chapter is a brief review of the endoscopic interventions currently performed in patients with acute or chronic pancreatitis. These interventions include biliary and pancreatic endoscopic sphincterotomy at the major or minor papilla, stricture dilatation on the common bile duct or main pancreatic duct, stent placement in the biliary or pancreatic ducts, stone extraction with or without extracorporeal shock wave lithotripsy, and transmural or transpapillary drainage of pancreatic fluid collections. As most of the studies reported were uncontrolled and retrospective, uncertainties persist with regard to the best approaches for treating the patients concerned. Appropriate patient selection, adequate expertise, and a supporting multidisciplinary infrastructure are essential prerequisites of a high success rate in improving the clinical condition of these patients. PMID- 15123091 TI - Ultrasound-guided endoscopic surgery. AB - Ten years' development of sectorial linear endoscopic ultrasound (EUS) has allowed us to perform guided biopsies of lymph nodes, mediastinal masses and pancreatic tumours. Furthermore, biopsy guided by EUS has been the first step in the development of interventional EUS. This development, in turn, has been made possible by the appearance of the interventional echoendoscope with a large working channel. EUS-guided biopsy obtains the best results for lymph nodes, anastomotic relapses and extrinsicz compression as well as for pancreatic tumours. Results in the literature show a global sensitivity of the technique that varies between 76 and 91%, a specificity of 84-100% and an accuracy of 78 94%. Using alcohol injection of the plexus nerves, the coeliac block guided by EUS is a simple technique. It will replace percutaneous access under US or CT scan guidance. Data from the literature show a significant reduction in pain of the order of 85-90% of cases-results that would have been good for percutaneous techniques. The technique of cystoenterostomy guided by EUS allows more accurate drainage of the cysts with a lower risk of perforation and haemorrhage. With regard to haemorrhage it should be emphasized that colour Doppler and power Doppler assessment of the punctures rules out the risk of vascular perforation during puncture but the risk of haemorrhage due to decompression of a vascular lesion in the vicinity of the cyst remains. The new ultrasound echoendoscope EG 38X, with a large working channel (3.8 mm), is now available. This system allows more effective drainage and also enables the placement of cystodigestive stents. PMID- 15123092 TI - Endoscopic treatment of strictures in Crohn's disease. AB - Patients with Crohn's disease often develop (recurring) intestinal stenosis. This is a result of continuous activation of fibrogenic cells by ongoing inflammation. Surgery is usually needed and consists of intestinal resection or strictureplasty. Medical therapy has not proven to be successful. Over the years endoscopic treatment has become more important. Uncomplicated stenosis, with a maximal length of 4 cm, can be treated by balloon dilatation. Indications, procedure and results are discussed. More recently, local corticosteroid injection in addition to balloon dilatation has been studied, but it remains to be seen whether long-term prevention of re-stenosis occurs. Other endoscopic therapies and new developments are also discussed in this chapter. PMID- 15123093 TI - Endoscopic stenting of colonic tumours. AB - Self-expandable metal stents (SEMS) are useful for the non-surgical relief of malignant colonic obstruction. They may be used both as a palliative measure and as a pre-operative bridge to facilitate a one-stage surgical resection of primary colonic tumours. SEMS may be placed endoscopically or by interventional radiologists without the use of endoscopy. In experienced centres SEMS can be successfully placed in approximately 90% of cases. Although it is known that the placement of these devices is feasible, there are no prospective trials comparing stent placement for colonic obstruction to routine surgical care. Additionally, there are no studies comparing the outcome of the method of placement (endoscopic versus radiological). This chapter reviews the types of expandable metal stent used for treatment of colonic obstruction, the indications for their insertion, their methods of insertion, and outcomes following insertion. Future research directions using expandable stents for colonic tumours are also addressed. PMID- 15123094 TI - [Can ART results be improved?]. PMID- 15123095 TI - [Embryo donation in France, in Europe and in the United States]. AB - Embryo donation, although proposed in a number of ART centres in the world is more often contemplated than performed, and very few publications report results on this subject. There is a great variability in program procedures and policies according to centres in the same country and between countries. In France, this activity is developing slowly. This is due to both the absence of information on the way to organise this activity at the medical level, and to legal constraints imposed by law. PMID- 15123096 TI - [Cystocele repair by vaginal approach with a tension-free transversal polypropylene mesh. Technique and results]. AB - OBJECTIVE: To evaluate mid term results, tolerance and efficiency of interposition of a tension-free polypropylene monofilament mesh by vaginal approach for the repair of cystoceles. PATIENTS AND METHODS: Fourty consecutive women underwent this new procedure between March 2001 and September 2002. After complete dissection of the cystocele, the polypropylene mesh (Gynemesh), Gynecare, Ethicon, Johnson & Johnson, France), proper cut-out was positioned under the bladder without any fixation. The lateral extensions of the mesh where introduced in the para-vesical spaces in contact with the arcus tendinus fascia pelvis. RESULTS: Before surgery all patients had cystocele of more or equal than grade II according to the POP-Q classification. Eight had grade II cystocele (20%) and 32 had grade III cystocele (80%). Thirty women had associated posterior or median prolapse (82.5%). Thirty women had urinary incontinence (75%). Mean age was 63.9 years (51-78). The procedure was performed without any difficulty in all cases. Duration of a cystocele cure procedure was 20 min +/- 2.2 (16-24). Mean follow-up is 16.4 months +/- 4.7 (12-24). The early complication rate was 7.5% (two vaginal erosions and one complete exposition of the mesh which requires secondary ablation). There was no mesh infection. The success rate was 95%. The satisfaction index overpasses 80% in 95% of our patients 6 months after the cystocele repair. DISCUSSION AND CONCLUSIONS: The interposition of a sub-vesical transversal tension-free polypropylene mesh by the vaginal route seems to be an excellent procedure in the definitive surgical treatment of grades II and III anterior vaginal wall prolapse. This new procedure is simple, mini-invasive, reproducible and efficient with low morbidity and good tolerance. PMID- 15123097 TI - [In utero transfer for preterm labor: experience of a regional perinatal hotline providing a 24-hour on call service]. AB - OBJECTIVE: Preterm labor is one of the major causes of concern for level I and II obstetricians. The purpose of this study was to determine the incidence of in utero transfer performed for preterm labor. We also aimed to evaluate the algorithm we used in case of call for preterm labor. This algorithm allowed us to study the rate of endovaginal sonography use prior to in utero transfer, to calculate its predictive value and to evaluate the risk of delivery during transfer. PATIENTS AND METHOD: We conducted an 8-months prospective study of all calls for preterm labor received at a regional call center in France (EU). All obstetrical data were entered in a computerized anonymous database. Three months after the first call midwives collected data from the receiving hospital. RESULTS: Calls for preterm labor account for 40% of calls for in utero transfer. Two hundred and sixty-five calls have been received for preterm labor; among them 50 cases were associated with a preterm rupture of membrane, a maternal or fetal pathology and 14 cases were lost for follow-up. Those 64 cases were excluded leaving 201 cases for analysis. Twenty-eight had a cervix dilated 4 cm, or more, while 173 had a cervix dilated less than 4 cm. Fifty percent of woman that had a cervical dilatation of 4 cm or more delivered more than 4 h after the call. Among the 173 patients that had a cervix dilated less than 4 cm, 71% had not delivered 7 days after the hotline call and 26% had an endovaginal ultrasonography performed before the transfer. None of the women that had a cervical length longer than 27 mm delivered in the 7 following days. None of the 176 women that were transferred delivered during the transfer. DISCUSSION AND CONCLUSION: In utero transfer for preterm labor is the leading cause of in utero transfer. Endovaginal ultrasonography prior to transfer should be performed in order to avoid unnecessary transfer. Women who have a preterm labor with a cervical dilatation of 4 cm or more are not an absolute contra-indication to in utero transfer. In those cases the transfer indication should be discussed on a case-to case basis including the actual term and the distance between hospitals. PMID- 15123098 TI - [Management of phyllodes tumors of the breast at the National institute of oncology of Rabat, Morocco]. AB - OBJECTIVES: Phyllodes tumors of the breast (PTB) are rare fibro epithelial tumors. Their terminology, histological classification and their treatment are exposed to controversy. The aim of our work is to underline the epidemiological, clinical, histological, therapeutical, prognostic and evolutive features of these tumours through a retrospective study and a review of the literature. PATIENTS AND METHODS: We reviewed at the National Institute of Oncology of Rabat, Morocco, between 1985 and 1998, all the files of patients with histological certainty of PTB, doubtful PTB were excluded. We collected epidemiological, clinical, histological, therapeutical, prognostic and evolutive features of these tumours. STATISTICS: quantitative parameters were represented by mean +/- S.D. and qualitative parameters by percentage or effective. RESULTS: We studied nine cases of PTB, which represented 0.09% of all primitive tumors of the breast treated at our institution during the study period. All our patients were female. Mean age was 37.3 +/- 10.07 years. Two of our patients (22.2%) had a history of fibroadenoma and 44.4% were nulliparous. Mean delay before consulting was 60.7 +/ 17.56 months and the median tumour size was 13 +/- 7.47 cm. Pathological findings were six benign or borderline phyllodes tumours (66.7%) and three cystosarcomas phyllodes (33.3%). The treatment consisted in simple mastectomy in seven cases (77.8%) and tumorectomy in two cases (22.2%). All the surgical margins were clear. Two of the three cystosarcomas phyllodes received adjuvant external bean radiation therapy 50 Gy on the thoracic chest wall. After a median outcome of 3 +/- 2 years ranging from 3 to 74 months, we did not note any relapse or metastasis. DISCUSSION AND CONCLUSION: In our series, PTB happened exclusively in females. History of fibroadenoma within 22.2% of the patients suggests the hypothesis of a filiation between these two entities. Their distinctive features were young age in diagnosis, long delay before consulting, important tumor size, predominant benign and borderline histological types, treatment mainly surgical and good local and distant control. PMID- 15123099 TI - [Conservation of human embryos in straws: safety in terms of human immunodeficiency virus 1]. AB - OBJECTIVE: The possibility of offering assisted reproductive technologies (ART) to HIV-positive couples has revived questions concerning the safety of the gametes and embryos cryopreservation in liquid nitrogen tanks. PATIENTS AND METHODS: We evaluated the safety of three types of straws - polyvinyl chloride (PVC), polyethylene terephthalate glycol (PETG) and so-called 'high-security' ionomeric resin (IR) - containing HIV-1 under standard conditions of cryopreservation. Potential HIV contamination was assessed by RT-PCR and then nested PCR. RESULTS: Under cryopreservation conditions, the sealed open ends of PVC and PETG straws were not safe. The ultrasound sealing system seems to be the weak link in obtaining total imperviousness of the straws. In contrast, both ends of the IR straws were safe for HIV in the framework of their use for ART. CONCLUSION: Sealing cryopreservation straws ultrasonically could incur the risk of not assuring their impermeability. Under standard cryopreservation conditions thermosealing of IR straws appears to be safe for HIV. PMID- 15123100 TI - [Pregnancy in rudimentary uterine horn: diagnostic and therapeutic difficulties]. AB - Ectopic pregnancy in a rudimentary uterine horn is extremely uncommon. Implantation of one embryo in the uterine cavity and of another in a rudimentary uterine horn is an extremely uncommon form of twin pregnancy. The authors report three cases of pregnancies in a rudimentary uterine horn. One was associated to a heterotopic pregnancy in the other eutrophic horn. Through these three cases, they report the risks incurred and the difficulties of the assumption of responsibility of this type of pathology, on the diagnostic as well as therapeutic level. But generally underline the interest of echography especially endovaginale and the coelioscopy in the early diagnosis of this type of uterine malformation. PMID- 15123101 TI - [Early isosexual precocious pseudopuberty revealing a juvenile granulosa cell tumor in a six-year-old girl]. AB - Juvenile granulosa cell tumors of the ovary are a rare form of neoplasm that makes up less than 5% of ovarian tumors in childhood and adolescence. About 90% are diagnosed in stage I (FIGO) with a favorable prognosis, whereas those at higher stages have a less favorable outcome. The authors describe a juvenile granulosa cell tumor expressed by an early pseudopuberty occurring in a 6-year old child. Clinically, an endocrine syndrome was associated with a pelvic mass. Hyperoestrogenia and serum alphafoetoprotein level were biologically detected. Tumor was localized strictly to the ovary, so conservative surgery was applied and proved sufficient to remove all tumor tissue. Histological examination showed typical microscopic aspect of a juvenile granulosa cell tumor. The patient is well 14 years after surgery with normal growth and mental development. PMID- 15123102 TI - [Sexual perversions or sexual variations?]. AB - Beyond the usual perversions Freud described and psychiatric nosography, some new sexual behaviours are nowadays coming to light, with a view to overstepping limits, and this in a mood for provoking, if not transgressing! As with classic perversions, the very play, the ambiguity with sexual difference, seems to be the very game and not the classic denial of this difference. PMID- 15123103 TI - [Intractable postpartum haemorrhages: where is the place of vascular ligations, emergency peripartum hysterectomy or arterial embolization?]. AB - OBJECTIVE: Update of knowledge on the various methods of management of intractable postpartum haemorrhage. METHOD: PubMed, MEDLINE were the electronic sources, in English and French languages, used for data retrieval. Uterine atony and abnormal placental insertions (placenta praevia or accreta) are the major causes of primary postpartum haemorrhages. To preserve fertility, we dispose of angiographic selective embolization or surgical vascular ligations. Embolization is a non-invasive method practicable by simple catheterization under local anesthesia. Vascular ligations of the uterine vessels or internal iliac arteries require mostly laparotomy. New and easier surgical methods, such as uterine compression or hemostatic suturing techniques have been described for which we lack experience. RESULTS: For uterine atony, the success rate of arterial embolization and uterine artery ligations is close to 100%. Ligation of internal iliac arteries is a little less effective and technically more difficult to carry out. It remains interesting in obstetrical traumatic hurts, which do not concern the uterus. If bleeding from the lower segment occurs during caesarean section, low uterine artery ligatures are necessary. These methods are all the more effective than they are prematurely implemented before the rise of major coagulopathy. In this case, uterine devascularization has also to be applied to ovarian vessels. With placenta accreta, accreta portion of the placenta can be left in place and arterial embolization or vascular ligations can be done. Nevertheless the main cause of failure with conservative treatments is placenta accreta. CONCLUSION: The simplest and the least morbid methods must be retained. After vaginal birth, arterial embolization can be done, if there is no maternal haemodynamic disorder nor interventional vascular radiology unit nearby. During caesarean section, progressive uterine artery ligation can be done adapted to the bleeding cause. In case of failure of a conservative treatment, it would be dangerous to multiply techniques. Emergency peripartum then should remain the choice procedure. PMID- 15123104 TI - [Utilization of PET scan in breast and gynaecologic cancers]. AB - The use of fluorine-18 fluorodeoxyglucose (FDG) in positron emission tomography (PET) is a major advance in imaging in the recent past. Its use in breast and gynaecologic cancers is not well established and needs to be discussed for each patient. This discussion must rely on a better knowledge of potential interests as well as of the limits of metabolic or logistic limits of PET. PMID- 15123105 TI - [How I perform...3rd and 4th degree obstetric analsphincter lacerations repair]. PMID- 15123106 TI - [How I perform...a transient ovariopexy]. PMID- 15123107 TI - [how I do the resection of the diaphragmatic peritoneum in ovarian carcinomatosis]. PMID- 15123108 TI - [Cesarean section on maternal request: pros]. PMID- 15123109 TI - [2003 update of recommendations for clinical practice: standards, options and recommendations for the use of FDG-PET in the management of gynaecological and breast cancers]. PMID- 15123110 TI - [Ultrasonic diagnosis of chorionicity in multiple pregnancies]. PMID- 15123111 TI - [Ultrasonic diagnosis of chorionicity in multiple pregnancies]. PMID- 15123113 TI - [Characteristics and use of the day after pill in France]. PMID- 15123114 TI - [Diagnostic smear: thin smear suspension collection. New method: analysis of results]. PMID- 15123115 TI - [What specialists will maternity services need tomorrow?]. PMID- 15123116 TI - [Long-term risks of polycystic ovaries syndrome]. AB - The prevalence of the polycystic ovaries syndrome (PCOS) is high but it is a heterogeneous disorder with implications in numerous medical domains. Abdominal obesity and insulin resistance, which are the main metabolic disorders, are strong links between hormonal abnormalities and long-term medical consequences. The latter begin to be better understood. Some studies suggest that PCOS may increase the risk for several conditions, including type 2 diabetes, dyslipidemia, hypertension, cardiovascular disease and some cancers. PMID- 15123117 TI - [Surgical management of rectovaginal septum endometriosis from a continuous series of 50 cases]. AB - OBJECTIVES: To assess the value of MRI and ano-rectal endosonography (ARES) for the diagnosis and surgical prognosis of rectovaginal septum endometriosis and to analyse the surgical management in order to evaluate its functional results and complications. PATIENTS AND METHODS: Retrospective study of 50 consecutive patients operated for a clinical presumption of endometriosis nodule of the recto vaginal septum. Thirty-nine patients had a MRI, 31 an ARES and 28 both exams. All the patients had a complete dissection of the rectovaginal septum and all lesions were excised. RESULTS: For the diagnosis of rectovaginal septum endometriosis nodule, MRI results are: sensitivity 73%, specificity 50%, positive predictive value (PPV) 89%, negative predictive value (NPV) 25%; for uterosacral ligaments involvement: sensitivity 84%, specificity 95%, PPV 94%, NPV 86% and for rectal wall infiltration: sensitivity 53%, specificity 82%, PPV 69%, NPV 69%. The ARES results for diagnosis of rectovaginal septum endometriosis nodule are: sensitivity 93%, specificity 100%, PPV 100%, NPV 50% and for rectal wall infiltration: sensitivity 100%, specificity 71%, PPV 81%, NPV 100%. ARES appeared more sensitive than MRI for the detection of rectal wall infiltration (P = 0.002) and for rectovaginal septum endometriosis nodule diagnosis (P = 0.03). Eighty nine percent of the patients had a coelioscopy in first intention and 15 laparoconversions were performed, 11 in order to perform a digestive resection: 45 nodules were found. In 43cases the nodule was excised, associated to 19 digestive resections, 30 colpectomys, and 22 uterosacral ligaments resections. Three patients required an additional surgical treatment by Hartman's procedure with Mickulicz's drainage for peritonitis. Forty-one nodules were endometriosis nodules: the two other cases were fibrosis nodules. Thirty-three patients were interviewed about the evolution of their pains over a mean history of 20 months: 90% of the patients were satisfied with the management results. DISCUSSION AND CONCLUSIONS: Our data support the efficiency of MRI for rectovaginal septum endometriosis nodule and uterosacral ligaments involvement diagnosis; accord ARES to rectovaginal septum endometriosis nodule diagnosis and its reliability in establishing a diagnosis of rectal wall involvement. The surgical cure of rectovaginal septum nodules without digestive infiltration is performed by coelioscopic or coelio-vaginal procedure, but in case of associated digestive affliction, laparotomy is actually the standard procedure in order to achieve a complete cure of the lesions. Complications, in particular peritonitis, are not frequent. Our data support the efficiency of radical surgical treatment for the improvement of pain symptoms. Results on fertility seem to be satisfactory, but complication risks suggest being careful in this indication. Clinical examination during a catamenial period is essential in order to evoke the diagnosis. MRI yields a complete map of the sub-peritoneal and peritoneal lesions and ARES allows for the diagnosis of an infiltration of the rectal wall. Pre-operative association of those two exams is actually indispensable for the surgical management of those patients, which consists of complete excision of endometriosical lesions and is efficient at treating pain symptoms and fertility. Complications are rare but severe, therefore, justifying a cure in specialised centres. PMID- 15123118 TI - [Thinking about the evolution of caesarean section rate at University Teaching Hospital of Dakar between 1992 and 2001]. AB - OBJECTIVE: The objective of this study is to answer the question: have we not been doing a lot of caesarean sections at University Teaching Hospital of Dakar? PATIENTS AND METHOD: This is an analytic study about caesarean section in 1992, 1996 and 2001; it was a prospective and longitudinal data collection from the epidemiological survey program carried through in Senegal about its obstetrical and surgical cover. For each year concerned, we have analysed caesarean section rate, maternal mortality rate and perinatal mortality rate. To eliminate the random part in observed variation, we used the comparison of proportions observed as a statistical test with a significant threshold less or equal to 5%. RESULTS: Caesarean section has gone from 12% in 1992 to 17.5 in 1996 and 25.2% in 2001. Operative indications are dominated by foeto-pelvic disproportion with an average of 31% and foetal suffering with an average of 25%. The increasing trend has been statistically significant for information's such as foeto-pelvic disproportion and maternal pathologies. The falling trend was statistically significant for indications in relation on relation to foetal suffering and scarred uterus. Gathering information has shown a stabilisation of "obligatory" caesarean rate around 41%, a decrease in "caution" caesarean rate from 50 to 37.2% and an increase in caesarean by "necessity" from 8.6 to 22.4%. The maternal mortality rate among women delivered has fallen from 1.4% to 0.8%, but postoperative surgery morbidity rate was still high around 10%, essentially due to infections. Reading of caesarean section rate has not a significant impact in perinatal prognosis. DISCUSSION AND CONCLUSION: Today there is an inflation of caesarean section at University Teaching Hospital of Dakar, without any significant loss of the maternal and perinatal mortality rate. The high level of complications due to surgery incite to reverse trends in order to get reasonable rate around 10 to 15% of childbirths. PMID- 15123119 TI - [Epidemiology of HPV infection]. AB - HPV DNA testing using Hybrid Capture 2 (Digene) was added to a program of liquid based (Thinprep, Cytyc) cervical cancer screening in a population of sexually active women aged from 20 to 62 years, without the history of uterine cervix pathology. 14.32% of 3832 women in this population were HPV positive. Positivity peaked in women aged 25-29 (19.4%) and gradually decreased, with 8% of positivity after 60 years. Positivity was independently related to parity, tobacco use and was correlated to cervical pathology. The rate of positivity in this population seems to preclude the use of HPV testing as a primary screening tool. PMID- 15123120 TI - [Puerperal uterine inversion: about two cases]. AB - Two cases of third- and second-degree acute puerperal uterine inversions that required surgical management after manual attempts failed are reported. The diagnosis was obvious in the first case but the second inversion was misdiagnosed as a myoma, which led to severe morbidity, linked with the hemorrhage. PMID- 15123121 TI - [Non heterosexual patients in gynaecology]. AB - Visible or invisible? Despite a growing tolerance of homosexuality over the past 30 years, little research on lesbian health or lesbian healthcare behaviour has been conducted in France. We have reviewed American studies in an attempt to investigate the relationship between sexual orientation and general and gynaecological health. In this paper, we stress the importance of trust in the doctor-patient relationship. PMID- 15123122 TI - [Viral transmission and medically assisted procreation: the Herpesviridae case]. AB - Procreation with sperm donation is at present achieved by insemination either in the uterus or in vitro, always from ejaculated and washed spermatozoa. Then, the infectious risk only exists if the donor sperm is capable of transporting the virus or its DNA, either by adhesion or by integration. With CMV, HSV1 and HSV2, medically assisted procreation in couples (AI or IVF-ET) does not increase the risk of viral contamination as compared with natural procreation, except possibly the cases of surgical procedure to pick up testicular sperm to be used in ICSI. Animal experiments show that, even if viral material is introduced in the oocyte, it may be eliminated from the embryo, at least for CMV. PMID- 15123123 TI - [Rapid microanalysis of lactate in cord blood and at fetal scalp blood sampling]. AB - Rapid microanalysis of lactate on fetal blood sampling is a new and useful method for fetal monitoring during labor, mainly because of the short lecture delay (1 min) and of the minimal necessary blood volume (5 microl). The lactatometer has the same size as a capillar glucometer and is not expensive. In comparison with pH, it has the advantage of determining metabolic acidosis. According to its excellent specificity in the diagnosis of fetal asphyxia, the lactate microanalysis is a useful complementary method for the fetal heart rate monitoring, the sensibility of which is well-known. A systematic measure of lactate in umbilical artery is an excellent means for self-education in the interpretation of fetal heart rate anomalies, and objective measure for evaluating the quality of intrapartum care and a well-suited defense against litigation, in combination with the 5 min Apgar score. PMID- 15123124 TI - [Is systematic iron supplementation justified during pregnancy?]. AB - Considering that young women have a high risk of iron store deficiency, the iron supplementation is largely proposed during pregnancy. However, a selective supplementation reserved to anaemia women, must be preferred to a systematic supplementation which improves biological parameters of mothers but have no effect on newborns. Iron is a potentially toxic element and a not justified, supplementation could expose to high iron level and to an oxidative stress which is also observed in pregnancy pathologies (preeclamptia, gestational diabetes). Furthermore a non controlled increase of erythrocyte mass by iron supplementation could also alter the placenta exchange. As a precaution, iron supplementation may be reserved to anaemia women or with high anaemia risk. For others, nutritional advises must permit to reach iron recommendation. PMID- 15123126 TI - [Evaluation of enlargement uteroplasty in the treatment of uterine anomalies following exposure to diethylstillbestrol (May 2003)]. PMID- 15123127 TI - [Routine use of ST-segment of fetal electrocardiogram for monitoring labor. A year's experience (preliminary results); Gynecol Obstet Fertil 2003; 31: 820 826]. PMID- 15123128 TI - [Two complications of ovarian drilling by fertiloscopy; Gynecol Obstet Fertil 2003; 31: 844-846]. PMID- 15123130 TI - [Instrumental extraction in 2002 in the "AURORE" hospital network: incidence and serious neonatal complications; Gynecol Obstet Fertil 2003; 31: 920-927]. PMID- 15123131 TI - [Perinatal issues: badly understood questions of public health!]. PMID- 15123132 TI - [Oocyte donation in France and national balance sheet (GEDO). Different European approaches]. AB - Oocyte donation (OD), a technique successfully used in Assisted reproductive techniques (ART), has been effective for 20 years as the palliative treatment of certain exocrine ovarian failures. Its indications have also been extended to cover some genetic diseases and documented occult ovarian insufficiency. In France, after an extensive judicial and ethical deliberation, OD was deemed legal and its practice was proscribed in the Bioethics Laws 94-653 and 94-654 of 29 July 1994. Its essential underlying principles are: voluntary, free, anonymous and confidential OD; according to a French decree to meet safety concerns, OD derived embryos must be quarantined for six months. In vitro fertilization (IVF) programs using donated oocytes are conducted in authorized centers under the responsibility of physicians authorized to retrieve and manipulate donated oocytes. The managers of these programs are the system's keystone, as their role includes, among other, the verification of indications, legal documents, public health safety, anonymous pairing of donor and recipient, and the collection of outcome data. At present, depending of the year, French programs have achieved 17 22% clinical pregnancy rates from transferred OD-derived frozen-thawed embryos, and follow-up studies reported excellent results concerning family relationships, and the physical and psychological development of the children. However, difficulties encountered in the daily practice of OD have led concerned physicians to form the Study group for OD (GEDO) to explore different topics with the aim of improving the overall management of this form of ART. We discussed these points herein, in comparison with the approaches of other European countries. PMID- 15123133 TI - [Does adenocarcinoma of uterine cervix have a worse prognosis than squamous carcinoma?]. AB - OBJECTIVE: The objective of this study was to examine the influence of histology on the outcome of patients with cervix carcinoma, treated with radiotherapy and radical surgery. PATIENTS AND METHODS: Clinical, histological, therapeutical and outcome data of 360 patients with stage IB-II cervix carcinoma patients (45 adenocarcinomas and 315 squamous cell carcinoma) managed between 1985 and 1998 were collected from the database of the Institut Gustave-Roussy. RESULTS: The incidence of adenocarcinomas slightly increased during the study period (P =0.07). Histological grade was higher for squamous cell carcinoma than for adenocarcinoma (P =0.08). Adenocarcinomas were smaller than squamous cell carcinoma (P =0.06). With only 38% of sterilized hysterectomy specimen vs 52% for squamous cell carcinomas (P =0.07), adenocarcinoma seemed to be less radiosensitive. With a median follow-up of 67 months, histological type did not influence survival. DISCUSSION AND CONCLUSIONS: Our study demonstrates that radiosensitivity is different between adenocarcinoma and squamous cell carcinoma of the cervix and that surgery may compensate the low radiosensitivity of adenocarcinoma. PMID- 15123134 TI - [Ultrasonographic measurement of cervical length in twin pregnancies with preterm labor: comparison with singleton pregnancies]. AB - OBJECTIVE: Our purpose was to assess the utility of ultrasonographic measurement of cervical length for predicting preterm delivery in pregnant women with twin pregnancy and preterm labor. PATIENTS AND METHOD: Prospective study in 192 singleton and 66 twin pregnancies with preterm labor before 34 weeks. Vaginal sonography for measurement of cervical length, and screening for fetal fibronectin in vaginal smears were performed at admission. The incidence of delivery before 34 weeks was compared with cervical length and the presence of fetal fibronectin. RESULTS: In singleton pregnancies, delivery before 34 weeks was significantly more frequent in women with a cervical length <30 mm (26.6 vs. 2.0%; P = 0.0004), or presence of fetal fibronectin (27.5 vs. 6.0%; P = 0.001). The corresponding negative predictive values (NPV) were 98% and 94%, respectively. In twin pregnancies, receiver characteristic curve analysis showed that a cervical length of 20 mm had the highest diagnostic performance in predicting preterm delivery. The corresponding NPV was 63%. In this group, delivery before 34 weeks showed no significant difference in women with a cervical length < 30 mm (43.7 vs. 38.8%), cervical length < 20 mm (52.0 vs. 36.6%, P = 0.219), or presence of fetal fibronectin (47.6 vs. 21.7%, P = 0.0705). DISCUSSION AND CONCLUSION: Ultrasonographic measurement of cervical length does not predict preterm delivery in twin pregnancies with preterm labor. PMID- 15123135 TI - [Bilateral breast cancer. Incidence and risk factors]. AB - OBJECTIVE: To clarify, thanks to a retrospective study of 24 bilateral breast cancer cases, the frequency, the risk factors and the prognosis of bilateral breast cancers. PATIENTS AND METHOD: Between 1984 and 1999, out of 506 patients treated for unilateral non-metastatic breast cancer at Gynecologic and Obstetric ward, at Maternity Souissi of Rabat, 24 cases of bilateral breast cancers were diagnosed. Our results were compared to those of the literature. RESULTS: The frequency of bilateral breast cancers was 4.7% (24/506). In 87.5% of cases, these were metachronous cancers with a mean interval of 45 months (12-144 months). Patients under 40 at first cancer ran a fivefold superior risk than women more than 40 (P < 0.05). In cases of T3 or T4 tumors, the risk was 10-fold superior to that in smaller ones (P < 0.05). DISCUSSION AND CONCLUSION: Significantly more first metachronous tumors were invasive adenocarcinoma cancers. Histologic type of first and second tumor was the same in all cases. The prognosis depends at once on the first and second cancer staging and the treatment must be done according to the same rules as in the first cancer. PMID- 15123136 TI - [Abnormal bacterial colonisation of the vagina and implantation during assisted reproduction]. AB - OBJECTIVE: To evaluate the efficiency of our treatment of vaginal infection for couples included in an IVF program. PATIENTS AND METHODS: Microbiologic screening of vaginal flora and semen has been performed one month prior to in vitro fertilization for 951 couples in 2000. Antibiotic treatment was prescribed in case of positive culture. RESULTS: Positive microbial growths were observed from endocervical and vaginal cultures in 218 women (22.9%). The clinical pregnancy rate was 30.29% in the group of patients without growth and 30.27% in the group with positive microbial growth. The implantation rate was significantly diminished in case of bacterial growth: 14.6 compared to 19.3% (P <0.02) for sterile endocervical culture. Five main bacterial species were found at the cervical level: Candida albicans (69 cases), Ureaplasma urealyticum (49 cases), Gardnerella vaginalis (43 cases), Streptococcus B or D (24 cases) and Escherichia coli (22 cases). Positive cultures from both vagina and semen were observed for 77 couples whose clinical pregnancy rate was 19.5 vs 36.2% in case of vaginal infection alone (P <0.01) with a spontaneous miscarriage rate of 46.7 compared to 17.6% (P <0.01). DISCUSSION AND CONCLUSION: Endocervical microorganisms, even treated with adapted antibiotics, may affect embryonic implantation. Positive culture from both female and male partner may enhance this negative effect. In this case, the best strategy would be to cancel the IVF treatment. PMID- 15123137 TI - [Uterine rupture following hysteroscopic surgery. A case report]. AB - A case of uterine rupture at 32 weeks' gestation, one year after hysteroscopic resection of uterine septa, is reported. A review of the literature shows only 16 other reported cases of uterine rupture following hysteroscopic surgery. The physiopathology and the prevention of these adverse effects are discussed. PMID- 15123138 TI - [Prenatal diagnosis of a probable human chimera after fertilization in vitro]. AB - Chimerism is the coexistence of more than one cell line in an individual, due to the fusion of originally separate zygotes. It has been very rarely described in humans. A 36-year-old woman referred for in vitro fertilization (IVF) had three embryos transferred leading to a monofetal pregnancy. Ultrasound examination at 17 weeks showed severe intrauterine growth retardation. Amniocentesis revealed a mixture of 46,XY and 46,XX clones. Histopathologic examination showed a dysmorphic fetus with female phenotype and severe growth retardation. Fusion of two of the three embryos (one male and one female) seems to be the most probable mechanism that could explain both cytogenetic and histopathologic observations. PMID- 15123139 TI - [Model of short-term psychological intervention in psychosomatic gynaecology]. AB - This article proposes a rapid psychological intervention model in psychosomatic gynaecology. The work draws from the method developed by Dr H. Davanloo (Intensive Short Term Dynamic Psychotherapy). First it consists in identifying and clarifying the defence mechanisms, second in exercising pressure on them. This pressure causes an increase in anxiety, an intensification of the defence mechanisms and the development of an intrapsychic crisis that induces emotions and painful feelings linked to past traumata. This activation of the unconscious can activate somatic symptoms (pain, unconscious movements, tics, muscular tensions) that highlight the link between the physical and psychic aspects. This work allows a rapid access to the painful emotions that turn to symptom. It indicates the therapeutic intervention zones and levels. It allows translating psychic reality in a simple, fast and efficient way. It brings heightened consciousness and comprehension for the therapist and the patient. PMID- 15123140 TI - [Strategies of antenatal diagnosis and management of pathologies related to multiple pregnancies]. AB - In the first trimester, ultrasound is crucial to screen for aneuploidies based on nuchal translucency, to diagnose major birth defects, and to establish chorionicity. Indeed prenatal diagnosis strategies as well as obstetrical management options are largely based on placental type. In the second trimester, maternal serum screening is not as effective, and ultrasound screening is more difficult in twins than in singletons. When a severe abnormality in found in one twin, selective termination of pregnancy may be considered. This technique is safe in dichorionic twins, but hazardous in monochorionic pregnancies. Selective termination in dichorionic twins is safer in the first trimester, underscoring the need for early prenatal diagnosis in twins. Monochorionic twin pregnancies carry specific risks, such as the twin-to-twin transfusion syndrome, which can be treated by endoscopic photo-coagulation of intertwin anastomoses in the severe early onset cases, or by amnioreduction in milder cases. PMID- 15123141 TI - [Contribution of VLPP (Valsalva leak point pressure) in the urodynamic assessment]. AB - The Valsalva leak point pressure (VLPP) is fully part of urologic assessment. Several factors can influence it: the patient's position, the bladder volume, the presence or absence of the urethral catheter, and the way of observing the leakage (direct visualisation or video-urodynamics). Sometimes VLPP is not feasible because patients cannot push strong enough. Nevertheless, a standardisation of the methodology has been suggested by the Societe internationale francaphone d'urodynamique (SIFUD). Values below 60 cmH2O should be considered suggestive for sphincter insufficiency. Conversely, the diagnosis of urethral sphincter insufficiency should rely on the tripod: clinical examination (low urethral mobility), low closure pressure and low VLPP. PMID- 15123142 TI - [Indications for Herceptin in breast cancer treatment]. AB - Trastuzumab (Herceptin) is the first clinically available oncogene-targeted therapy to produce a significant survival advantage in advanced breast cancer. This monoclonal humanized antibody is indicated as a single-agent second-line or third-line treatment or in combination with paclitaxel for metastatic breast cancer with HER2 gene amplification or high overexpression of HER2 (3+ on immunohistochemistry). Assessment with regular monitoring of the left-ventricular function during therapy is necessary to detect and manage potential cardiac dysfunction. International randomized trials in the adjuvant setting are ongoing. PMID- 15123143 TI - [For the routine induction of labor at 41 weeks]. PMID- 15123144 TI - [Emergency contraception in France: the user profile. Gynecol Obstet Fertil 2003; 31: 724-729]. PMID- 15123146 TI - [Cesarean section in the Burgundy perinatal network: reasons for differences in the management of mothers. Gynecol Obstet Fertil 2003; 31: 606-613]. PMID- 15123147 TI - The bright future of coronavirology. PMID- 15123148 TI - Risk factors for feline coronavirus seropositivity in cats relinquished to a UK rescue charity. AB - Two thousand, two hundred and seven cats from 14 shelters of a major UK cat charity were blood tested for feline coronavirus (FCoV) antibodies. Data was collated on breed, sex, age, number of cats at original location, outdoor access, health status, and time spent in the shelter prior to sampling (range 0 to 4 years). Some cats were also tested for feline leukaemia virus antigen, feline immunodeficiency virus, and Toxoplasma gondii antibodies. The effect of these variables on FCoV seropositivity was explored by multivariable logistic regression. FCoV seropositivity in cats that had spent 5 days or less in a shelter at sampling was significantly associated with a multi-cat origin, cats aged 3 years or less, and Persian breed. Whether pet, stray or feral, health status, indoor/outdoor access, and sex had no significant effect. Overall FCoV seropositivity was associated with time spent in a shelter but this association was not linear. Cats that had spent more than 60 days in a shelter were over five times as likely to be seropositive. This may be the result of a change in husbandry from solitary to communal housing for cats remaining in shelters long term. Rescue of cats for less than 60 days was not associated with a significant increasing risk of seropositivity. Significant variation existed in seropositivity between individual shelters overall and in cats rescued for less than 5 days. These findings may reflect inter-shelter variation in cat husbandry and variation in seropositivity of shelter intake respectively. PMID- 15123149 TI - Feline leucocyte antigen class II polymorphism and susceptibility to feline infectious peritonitis. AB - There are four outcomes to feline coronavirus (FCoV) infection: the development of feline infectious peritonitis (FIP, which is immune-mediated), subclinical infection, development of healthy lifelong carriers and a small minority of cats who resist infection (Addie and Jarrett, Veterinary Record 148 (2001) 649). Examination of the FCoV genome has shown that the same strain of virus can produce different clinical manifestations, suggesting that host genetic factors may also play a role in the outcome of infection. FIP is most prevalent amongst pedigree cats, although how much of this is due to them living in large groups (leading to higher virus challenge and stress which predisposes to FIP) and how much is due to genetic susceptibility is not known. If host genetics could be shown to play a role in disease, it may allow the detection of cats with a susceptibility to FIP and the development of increased population resistance through selective breeding. The feline leucocyte antigen (FLA) complex contains many genes that are central to the control of the immune response. In this preliminary study, we used clonal sequence analysis or reference strand conformational analysis (RSCA) to analyse the class II FLA-DRB of 25 cats for which the outcome of FCoV exposure was known. Individual cats were shown to have between two and six FLA-DRB alleles. There was no statistically significant association between the number of alleles and the outcome of FCoV infection. No particular allele appeared to be associated with either the development of FIP, resistance to FCoV, or the carrier status. However, the analysis was complicated by apparent breed variation in FLA-DRB and the small number of individuals in this study. PMID- 15123150 TI - Evaluation of an in-practice test for feline coronavirus antibodies. AB - A commercially available in-practice test for feline coronavirus (FCoV) antibodies (FCoV Immunocomb, Biogal Galed Laboratories) was evaluated by comparison with the gold standard FCoV immunofluorescent antibody (IFA) test. One hundred and three serum or plasma samples were selected and tested: 70 were positive by both tests, 24 were negative by both tests. The in-practice test produced five false positive and four false negative results. The sensitivity of the in-practice test was 95% and the specificity was 83%. When the titres were compared it was found that the in-practice test results were significantly correlated with IFA titres but the degree of correlation was not likely to be clinically useful. The IFA titres of the four false negative samples were found to be low (less than 40) which suggests that even a cat with a false negative result is still unlikely to be excreting FCoV. A negative result with the in practice assay is likely to be reliable for screening cats prior to entry into an FCoV-free cattery or stud. It would also be useful in the investigation of suspected FIP as most cats with this condition have high IFA titres of antibodies. A strong positive result would be useful in the diagnosis of FIP (in conjunction with other biochemical and cytological testing), but positive results would be of limited value in monitoring FCoV infection in healthy cats as the antibody titre could not be reliably compared with those obtained with IFA. All positive results obtained using the in-practice kit should be confirmed and titrated by IFA. The kit also appeared to work efficiently with ascites samples (n=6) but too few samples were analysed to draw firm conclusions. PMID- 15123152 TI - Common virus infections in cats, before and after being placed in shelters, with emphasis on feline enteric coronavirus. AB - The purpose of this study was to determine the origin and subsequent spread of feline calicivirus (FCV), feline herpesvirus (FHV), and feline enteric coronavirus (FECV) in cats relinquished to shelters. FCV was isolated from the oral fauces of 11% of healthy cats upon entry, and isolation rates were highest for kittens (33%). FHV shedding was very low (4%) at the time of entry and occurred mainly in juveniles. FECV shedding was also common among newly relinquished cats (33%), especially older kittens and juveniles (90%). The subsequent spread of all three viruses was rapid and efficient in the shelter environment. Fifteen percent of cats were shedding FCV, 52% FHV, and 60% FECV after 1 week. More detailed studies were done with FECV shedding, which could be accurately quantitated. The amounts of FECV shed by infected cats ranged from 10(2)to 10(16)particles/swab of feces. FECV shedding was several logs higher in young kittens with primary infection than adult cats with primary infections. The mean levels of FECV shedding among adults were the same for primary and chronic infections. Although shelters were not the primary source of these viruses for many relinquished cats, factors intrinsic to the shelter environment were critical in amplifying shedding and spread to susceptible individuals. Extrinsic factors were especially important for the spread of FHV and FECV. FHV shedding rates increased from 4% to 50% in 1 week's time. The speed and magnitude of the increase in FHV shedding suggested that there was reactivation of latent infections as well as acquisition of new infections. FECV shedding increased 10 to 1,000,000 fold in 1 week among cats that were already infected at entry, and more than one-half of initially negative cats were shedding FECV a week later. Feline calicivirus infection was the least likely to spread in the shelter. The infection rate only increased from 11 to 15% in 1 week. PMID- 15123151 TI - High viral loads despite absence of clinical and pathological findings in cats experimentally infected with feline coronavirus (FCoV) type I and in naturally FCoV-infected cats. AB - Specified pathogen-free cats were naturally infected with FCoV or experimentally infected with FCoV type I. Seroconversion was determined and the course of infection was monitored by measuring the FCoV loads in faeces, whole blood, plasma and/or monocytes. Tissue samples collected at necropsy were examined for viral load and histopathological changes. Experimentally infected animals started shedding virus as soon as 2 days after infection. They generally displayed the highest viral loads in colon, ileum and mesenteric lymph nodes. Seroconversion occurred 3-4 weeks post infection. Naturally infected cats were positive for FCoV antibodies and monocyte-associated FCoV viraemia prior to death. At necropsy, most animals tested positive for viral shedding and FCoV RNA was found in spleen, mesenteric lymph nodes and bone marrow. Both experimentally and naturally infected cats remained clinically healthy. Pathological findings were restricted to generalized lymphatic hyperplasia. These findings demonstrate the presence of systemic FCoV infection with high viral loads in the absence of clinical and pathological signs. PMID- 15123153 TI - Disease outcome and cytokine responses in cats immunized with an avirulent feline infectious peritonitis virus (FIPV)-UCD1 and challenge-exposed with virulent FIPV UCD8. AB - Eight cats were immunized with an avirulent strain of feline infectious peritonitis virus (FIPV)-UCD1, then challenge-exposed to a highly virulent cat passaged strain (FIPV-UCD8). Th1 and Th2 cytokine profiles in the peripheral blood mononuclear cells (PBMCs) were measured throughout in the experiment. No clinical signs of FIP were evident in the experimental cats after immunization. After challenge, the immunized cats demonstrated one of four clinical outcomes: (1) classical effusive FIP; (2) accelerated FIP; (3) non-effusive FIP, or (4) resistance to challenge. Only minor cytokine changes were observed following immunization, however, several cytokine changes occurred following challenge exposure. The most noteworthy changes were in tumor necrosis factor-alpha (TNF alpha) and interferon gamma (IFN-gamma) levels. Our preliminary findings suggest that immunity against FIP is associated with TNF-alpha and IFN-gamma response imbalance, with high TNF-alpha/low IFN-gamma mRNA responses favouring disease and low TNF-alpha/high IFN-gamma mRNA responses being indicative of immunity. PMID- 15123154 TI - Tissue distribution of a feline AGP related protein (fAGPrP) in cats with feline infectious peritonitis (FIP). AB - Feline alpha(1)-acid glycoprotein (fAGP) increases during feline infectious peritonitis (FIP). We have recently identified a 29 kDa protein that we named feline AGP-related protein (fAGPrP) due to its cross-reactivity with an anti human AGP monoclonal antibody. In this work we describe the tissue distribution of fAGPrP during FIP, and its relationship with feline coronavirus (FCoV) and myeloid cells. Tissues from five control cats and from 15 cats with FIP were examined by immunohistochemistry using monoclonal antibodies against human AGP, FCoV and myeloid antigens. Diffuse fAGPrP positivity within the lesions, likely due to vascular plasma leakage, endothelial and epithelial lining were detectable. Compared to controls, fAGPrP-expressing cells often increased in number and were diffusely distributed in lymph nodes, as usually occurs for IgM producing plasma cells during early immune responses. These findings did not depend on the presence of FCoVs or of myeloid cells, suggesting that fAGPrP is not directly involved in the pathogenesis of FIP. PMID- 15123155 TI - Use of recombinant feline interferon and glucocorticoid in the treatment of feline infectious peritonitis. AB - A total of 12 clinically ill cats previously diagnosed as feline infectious peritonitis (FIP) were treated with a combination of recombinant feline interferon and glucocorticoid. A complete remission (over 2 years) and a partial remission (2 to 5 months) were observed in four (33.3%) and four (33.3%) cases, respectively. Those that survived for more than 2 years were all older cats (6 to 16 years old) with the effusive form of FIP. PMID- 15123156 TI - Modified vaccinia virus Ankara as a vaccine against feline coronavirus: immunogenicity and efficacy. AB - Feline infectious peritonitis virus (FIPV) is a coronavirus that induces a fatal systemic disease mediated by an inappropriate immune response. Most previous vaccination attempts against FIPV were unsuccessful because IgG antibodies against the surface protein enhance the infection. However, two studies have shown that poxvirus vectors (vaccinia WR and canarypox) expressing only the FIPV membrane (M) protein can elicit a partially protective immunity which is supposed to be cell-mediated (Virology 181 (1991) 327; International patent WO 97/20054 (1997)). In our study, we report the construction of another poxvirus, the modified vaccinia virus Ankara (MVA), as an expression vector for the FIPV M protein. In this vector, the M gene has been inserted downstream a strong early/late promoter, whereas the two previously described poxviruses expressed the M protein during their early stage only. The immunogenicity of the recombinant MVA-M was evaluated in the murine model which revealed an effect of the vector on the Th1/Th2 balance. The vaccine was then tested in cats to evaluate its efficacy in an FIPV 79-1146 challenge. Vaccinated kittens developed FIPV-specific antibodies after immunization, however, none of them was protected against FIPV. Our results suggest a crucial role for the type of poxviral promoter that must be used to induce an effective immune response against FIPV. PMID- 15123157 TI - FIP: a novel approach to vaccination. Proceedings from the 2nd International FCoV/FIP Symposium, Glasgow, 4-7 August 2002. AB - Feline infectious peritonitis (FIP) is a fatal disease of cats. Early attempts at vaccination have been unsuccessful, some even serving to exacerbate the disease through antibody-dependent enhancement. Replication-incompetent feline foamy virus (FFV) transducing vectors are being developed as potential vaccine agents, into which immunogenic fragments of feline coronavirus (FCoV) proteins will be inserted. To use a recombinant viral vector to express FCoV proteins, the agent chosen should be apathogenic and replication incompetent within the host following gene delivery. Spumaviruses confer several advantages over the more traditionally explored retroviral vectors. Stable helper cell line clones have been established by transfection of CRFK cells with FFV tas and assessed using beta-galactosidase assays, PCR, immunofluorescence and western blotting. The generation of infectious virions using these cell lines has been investigated using tas-deleted FFV vectors containing the enhanced green fluorescent protein (eGFP) cassette. PMID- 15123158 TI - Recommendations from workshops of the second international feline coronavirus/feline infectious peritonitis symposium. AB - In August 2002, scientists and veterinarians from all over the world met in Scotland to discuss feline coronavirus (FCoV) and feline infectious peritonitis (FIP). The conference ended with delegates dividing into three workshops to draw up recommendations for FCoV control, diagnosis and treatment and future research. The workshops were chaired by the three authors and the recommendations are presented in this paper. PMID- 15123159 TI - Human severe acute respiratory syndrome (SARS) and feline coronaviroses. PMID- 15123160 TI - Commentary for special issue on feline behavior. PMID- 15123161 TI - Thinking outside the box: feline elimination. AB - Feline elimination problems are the leading behavioral complaint of cat owners. When presented with a feline elimination problem there are three main diagnostic categories: medical problems, marking or toileting problems. It is important to first perform diagnostic tests to rule out and/or address underlying medical issues and all feline house soiling patients should receive a comprehensive physical examination. When the elimination problem persists after a medical problem has either been ruled out or remedied, a behavioral diagnosis should be obtained. The primary distinction that must be made in a behavioral diagnosis is whether the cat is engaging in marking behavior or selecting a spot other than the litterbox for elimination (a toileting problem). The motivation for urine marking may be territorial behavior or anxiety/stress whereas toileting problems are often triggered by medical causes, aversions, preferences or anxiety. Marking animals should be neutered and additional treatment measures may include reducing conflict and stress in the environment. Drug therapy has been long used to help control urine marking and recent studies have furthered our knowledge about the most appropriate treatments. Treatment for toileting problems should focus on providing an attractive litterbox while reducing the attractiveness or accessibility of inappropriate target spots. PMID- 15123162 TI - Fractious cats and feline aggression. AB - Despite variation in terminology it is clear that the context in which feline aggression occurs is important for classification, prevention and treatment. Many of the aggressions discussed can be considered variants of normal feline behavior, so client education is particularly important. Educated clients will know when their cat can benefit from veterinary intervention, and when closing a door may be sufficient. The goals of treating all feline aggressions should be safety for all concerned, and quality of their living environment for the cats. The greater our knowledge, the more likely we are to achieve these goals. PMID- 15123163 TI - Social organization in the cat: a modern understanding. AB - An increasing body of research work has made it clear that, while Felis catus can survive in the solitary state, social groups with an internal structure, are formed whenever there are sufficient food resources to support them. Most people who have cats have two or more cats. Failure to understand what will promote either friendly or aggressive behavior can lead to various behavior problems, including aggression and conflict over resources, such as food, resting sites and litterboxes. An understanding of the natural social organization, relationships and communication between cats is therefore essential, and is the subject of this paper. PMID- 15123164 TI - Paradigms for pharmacologic use as a treatment component in feline behavioral medicine. AB - Veterinary behavioral medicine remains an under-supported, under-appreciated, and under-taught specialty within veterinary medicine. Neuropsychopharmacology is the aspect that has provided the field with the most scientific legitimacy, but is also one of the most hotly debated. Paradigms for use of pharmacologic intervention include firstly ruling out any underlying medical cause. If a behavioral diagnosis can be made, treatment with psychotropic medication may be considered, although their use is most effective as part of an integrated treatment program that includes behavior modification. Used without an understanding of the mechanism of action, pharmacologic intervention may only blunt or mask behavior without altering processes or environments that produced the behavior. This paper reviews specific drugs, mechanism of action of those drugs, and relevant uses are reviewed for cats. Future advances in treatment in veterinary behavioral medicine will be pharmacological and neurophysiological. As the field of veterinary behavioral medicine expands, its paradigm will enlarge to include routine combination therapy and the implementation of neuropharmacological intervention as a diagnostic tool. PMID- 15123165 TI - An ethical viewpoint: the role of veterinarians and behaviourists in ensuring good husbandry for cats. AB - Cat owners commonly consider their pets to be members of their families, and many factors contribute to a high level of owner attachment to their cats. Suppression of a cat's emotional needs in favour of the emotional requirements of the owner may produce a less satisfactory relationship for the owner, and usually for the cat as well. Owners' failure to comprehend their cat's true demands of life, and their false expectation of their ability to fulfil human psychological demands lies at the heart of many feline behaviour problems. PMID- 15123166 TI - Up date on IGFBP-4: regulation of IGFBP-4 levels and functions, in vitro and in vivo. AB - Of the six known high affinity insulin-like growth factor binding-proteins (IGFBPs), IGFBP-4 appears to be unique in that it is the only IGFBP that functions mostly like a traditional binding protein. In this regard, none of the IGF independent effects that have been ascribed for other IGFBPs have been described for IGFBP-4. However, recent in vitro and in vivo studies, in particular the recent identification of pregnancy-associated plasma protein-A as a major IGFBP-4 protease, are consistent with the idea that IGFBP-4 is an extremely important component of IGF system in several tissues including gonads and bone. In this review, we have provided an update on IGFBP-4 research and we have summarized our current understanding of the regulation of levels and actions of IGFBP-4 and proteolytic fragments both in vitro and in vivo. PMID- 15123167 TI - The phospholipase C-InsP3 pathway is involved in calcium mobilization induced by growth hormone in hepatocytes. AB - We investigated the effects of bovine GH (bGH) on Ca(2+) handling, phospholipase C (PLC) activation and inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] formation in hepatocytes. bGH generates oscillations in cytosolic free Ca(2+) concentration ([Ca(2+)](i)) in single male rat hepatocytes microinjected with the photoprotein aequorin. In the absence of extracellular Ca(2+) these transients persisted for more than 10 min indicating a requirement for intracellular Ca(2+). Treatment of the hepatocyte with the phosphatidylinositol-specific phospholipase C (PI-PLC) inhibitor U-73122 removed the oscillations. These results suggest bGH-induced oscillations are due to PLC activation and generation of Ins(1,4,5)P(3). We measured the mass of Ins(1,4,5)P(3) in freshly isolated hepatocyte suspensions in response to bGH, and vasopressin as a control. Both agonists rapidly increased the levels of Ins(1,4,5)P(3). This is the first study to indicate that early events in the signal transduction pathways mediated by GH in hepatocytes involve intracellular Ca(2+) mobilization via activation of a PI-PLC and subsequent Ins(1,4,5)P(3) production. PMID- 15123168 TI - The IGF-I system component concentrations that decrease with ageing are lower in obesity in relationship to body mass index and body fat. AB - The aim of this study was to investigate the GH-IGF-I axis in healthy adults and its relationship to obesity. We studied 268 subjects: 134 men and 134 women, and determined anthropometric and body composition variables. Serum total IGF-I was measured by radioimmunoassay, serum free IGF-I concentrations by enzyme linked immunosorbant assay and serum IGFBP3 concentrations by radioimmunoassay. In men, we observed a decrease in total IGF-I, free IGF-I and IGFBP-3 throughout decades. In women, the body mass index and fat mass were higher throughout decades, and we observed a similar decrease to that in men in total IGF-I, free IGF-I and IGFBP3. In men with obesity, as measured by body fat, free IGF-I concentrations were lower than those without obesity; in women with obesity, total IGF-I concentrations and free IGF-I concentrations were lower than in those with obesity. These changes were observed in relationship to obesity when the subjects were adjusted for differences in age. We showed that in controls randomly selected, the GH-IGF-I axis component concentrations that decrease with increasing age are lower in obesity, especially in women, and that this decrease is related to body mass index and body fat. PMID- 15123169 TI - The effect of temperature on juvenile Mozambique tilapia hybrids (Oreochromis mossambicus x O. urolepis hornorum) exposed to full-strength and hypersaline seawater. AB - The effects of temperature on the salinity tolerance of Mozambique-Wami tilapia hybrids (Oreochromis mossambicus x O. urolepis hornorum) were investigated by transferring 35 g/l, 25 degrees C-acclimated fish to 35, 43, 51 or 60 g/l salinity at 15, 25 or 35 degrees C for 24 h, and by assaying gill tissue for branchial Na(+), K(+)-ATPase activity at the three temperatures after acclimating the fish to 15, 25 or 35 degrees C for 2 weeks. Tilapia survived all salinities at 25 and 35 degrees C; however, at 15 degrees C, mortality was 85.7% and 100% in the 51 g/l and 60 g/l groups, respectively. There was a significant interaction between temperature and salinity, as plasma osmolality, [Na(+)] and [Cl(-)] were significantly increased at 51 and 60 g/l salinity in 35 degrees C water (P<0.001). Additionally, muscle water content was significantly reduced at 43 g/l, 15 degrees C relative to pre-transfer values (P<0.001). Branchial Na(+), K(+)-ATPase activity was reduced at 15 degrees C regardless of acclimation temperature, and 25 degrees C-acclimated gill tissue did not show an increase in activity when assayed at 35 degrees C. Results indicate that the effects of a combined temperature-salinity transfer on plasma osmolality and ion concentrations, as well as muscle water content, are greater than when either challenge is given alone. Additionally, branchial Na(+), K(+)-ATPase activity is altered when assayed at varying temperatures; in the case of 15 degrees C, regardless of acclimation temperature. Our enzyme activity data may indicate the presence of a high temperature isoform of branchial Na(+), K(+)-ATPase enzyme. PMID- 15123170 TI - Maturation-related variations in prostaglandin and fatty acid content of ovary in the kuruma prawn (Marsupenaeus japonicus). AB - Total lipid, fatty acids and prostaglandins (PGF(2 alpha) and PGE(2)) in the ovary of kuruma prawns (Marsupenaeus japonicus) were measured during ovarian development. The level of ovarian total lipid increased with an increase in the gonad-somatic index (GSI). No significant difference was found in fatty acid composition among different stages of ovarian development. However, the content of arachidonic acid (precursor of PG(2)), but not eicosapentanoic acid (precursor of PG(3)), was significantly lower at stages I and II than at stage V (P<0.01). When ovarian PGF(2 alpha) and PGE(2) levels were plotted against GSI, no correlation was found in either PG. However, in terms of ovarian developmental stages, the level of ovarian PGs was high (approx. 20 pg/mg) at stage I, followed by marked decreases at stages IV and V (PGF(2 alpha), P<0.01) and stage IV (PGE(2), P<0.01). These results suggest that ovarian PGs and arachidonic acid are deeply involved in ovarian maturation in kuruma prawns. PMID- 15123171 TI - Basal metabolic rate, food intake, and body mass in cold- and warm-acclimated Garden Warblers. AB - We address the question of whether physiological flexibility in relation to climate is a general feature of the metabolic properties of birds. We tested this hypothesis in hand-raised Garden Warblers (Sylvia borin), long-distance migrants, which normally do not experience great temperature differences between summer and winter. We maintained two groups of birds under cold and warm conditions for 5 months, during which their body mass and food intake were monitored. When relatedness (siblings vs. non-siblings) of the experimental birds was taken into account, body mass in cold-acclimated birds was higher than in warm-acclimated birds. BMR, measured at the end of the 5-month temperature treatment, was also higher in the cold- than the warm-acclimated group. Migrant birds thus seem to be capable of the same metabolic cold-acclimation response as has been reported in resident birds. The data support the hypothesis that physiological flexibility is a basic trait of the metabolic properties of birds. PMID- 15123172 TI - Milk production and composition in reindeer (Rangifer tarandus): effect of lactational stage. AB - Milk yield and composition of major milk constituents were measured in captive, nursing reindeer. Registration of milk production was performed during two successive lactations (2001 and 2002). The milk yield was significantly affected by week of lactation (P<0.001) and by individual (P<0.001). The lactation curve had an asymmetrical peak 3 weeks postpartum and the milk yield at peak lactation was 983 g/day (range 595-1239). The length of lactation varied from 24 to 26 weeks and average total milk production was 99.5 kg. From peak lactation the milk production decreased linearly (P<0.001) until milk production was terminated. Mean values for content of major milk constituents were 15.5% fat, 9.9% protein and 2.5% lactose. The content of fat and protein increased markedly with the lactation stage (P<0.001), while lactose showed a slight decrease (P<0.001). The milk composition was significantly affected by stage of lactation (P<0.001). There was a marginally significant decrease in protein:fat ratio (P=0.06) as protein was substituted by fat with stage of lactation. The caloric value of the milk averaged 8.7 kJ/g and increased significantly with the stage of lactation (P<0.001). The overall increase in milk gross energy content during lactation was 67.6%. The energy output averaged 7996 kJ/day at peak lactation and decreased significantly during the course of lactation (P=0.002). PMID- 15123173 TI - Species specificity in avian sperm:perivitelline interaction. AB - The interaction of chicken spermatozoa with the inner perivitelline layer from different avian species in vitro during a 5 min co-incubation was measured as the number of points of hydrolysis produced per unit area of inner perivitelline layer. The average degree of interaction, as a proportion of that between chicken spermatozoa and their homologous inner perivitelline layer, was: equal to or greater than 100% within Galliformes (chicken, turkey, quail, pheasant, peafowl and guineafowl); 44% within Anseriformes (goose, duck); and less than 30% in Passeriformes (Zebra Finch) and Columbiformes (collared-dove). The homologue of the putative chicken sperm-binding proteins, chicken ZP1 and ZP3, were identified by Western blotting with anti-chicken ZP1/ZP3 antibody in the perivitelline layers of all species. The functional cross-reactivity between chicken spermatozoa and heterologous inner perivitelline layer appeared to be linked to known phylogenetic distance between the species, although it was not related to the relative affinity of the different ZP3 homologues for anti-chicken ZP3. This work demonstrates that sperm interaction with the egg investment does not represent such a stringent species-specific barrier in birds as it does in mammals and marine invertebrates. This may be a factor in the frequency of hybrid production in birds. PMID- 15123174 TI - Tectal responses to potassium loads and subsequent visual stimuli in the toad, Bufo bufo. AB - Calling male toads were tested behaviourally for their prey catching responses to wormlike stimuli and assigned to groups of non-hungry and hungry depending on their prey catching motivation before being prepared for visual unit, massed unit and slow potential shift (SPS) recording from the optic tectum. Control recordings to visual stimuli were made before recording the effects of application of isotonic solutions containing concentrations of 0-41 mM K(+). Application of solution was followed by presentation of the visual stimulus while the solution still bathed the tectum. The best tectal responses were made to large square visual stimuli in the non-hungry toads, perhaps because recordings were made in the breeding season. Responses of the tectum to solution addition were significant in the concentration range of 7-17 mM K(+). Hungry toads showed an earlier, smaller response than non-hungry (sexually motivated) animals. When the visual stimulus was presented, there were unit and massed unit responses at all bathing solution concentrations, which were larger in non-hungry animals. These experiments revealed that toads motivated to feed respond earlier than non hungry toads to application of artificial CSF to the tectum, though non-hungry toads responded best to the subsequent visual stimulus. PMID- 15123175 TI - Hysteresis of heart rate and heat exchange of fasting and postprandial savannah monitor lizards (Varanus exanthematicus). AB - Reptiles are ectothermic, but regulate body temperatures (T(b)) by behavioural and physiological means. Body temperature has profound effects on virtually all physiological functions. It is well known that heating occurs faster than cooling, which seems to correlate with changes in cutaneous perfusion. Increased cutaneous perfusion, and hence elevated cardiac output, during heating is reflected in an increased heart rate (f(H)), and f(H), at a given T(b), is normally higher during heating compared to cooling ('hysteresis of heart rate'). Digestion is associated with an increased metabolic rate. This is associated with an elevated f(H) and many species of reptiles also exhibited a behavioural selection of higher T(b) during digestion. Here, we examine whether digestion affects the rate of heating and cooling as well as the hysteresis of heart rate in savannah monitor lizards (Varanus exanthematicus). Fasting lizards were studied after 5 days of food deprivation while digesting lizards were studied approximately 24 h after ingesting dead mice that equalled 10% of their body mass. Heart rate was measured while T(b) increased from 28 to 38 degrees C under a heat lamp and while T(b) decreased during a subsequent cooling phase. The lizards exhibited hysteresis of heart rate, and heating occurred faster than cooling. Feeding led to an increased f(H) (approximately 20 min(-1) irrespective of T(b)), but did not affect the rate of temperature change during heating or cooling. Therefore, it is likely that the increased blood flows during digestion are distributed exclusively to visceral organs and that the thermal conductance remains unaffected by the elevated metabolic rate during digestion. PMID- 15123176 TI - Localization, morphology and function of the mitochondria-rich cells in relation to transepithelial Na(+)-transport in chicken lower intestine (coprodeum). AB - The correlation between morphology of the mitochondria-rich cells (MR cells) in chicken lower intestine, coprodeum, and dietary sodium levels, has been investigated, using hens with differing dietary intake of NaCl and plasma aldosterone levels. Additionally, the function of the MR cells was evaluated in relation to proton secretion/exchange. Epithelium from the coprodeum was examined by optical, transmission and scanning electron microscopy, and Na(+)-transport across the coprodeal epithelium was measured electrophysiologically in Ussing chambers. To investigate the function of MR cells, lectin-, enzyme- and immunohistochemistry methods were used. The MR cells were generally located in the epithelium on the upper parts of the sides of mucosal folds. Long microvilli, high but variable toluidine blue affinity/electrondensity and numerous mitochondria were the main features distinguishing them from the surrounding epithelial cells. Two main MR cell types were observed, differing in microvillous morphology, diameter and toluidine blue affinity/electrondensity. This probably reflected differences in maturity and activity. The MR cells expressed a positive carbonic anhydrase reaction and a proton exchange similar to the absorptive intestinal epithelial cells, but exhibited no specific demonstrable proton secretion. A close correlation between the ultrastructure of the MR-cells, dietary sodium levels, plasma aldosterone and transepithelial Na-transport was observed. PMID- 15123177 TI - Nutritional condition and serum biochemistry for free-living Swainson's Hawks wintering in central Argentina. AB - We assessed the nutritional condition and established reference values for serum chemistry parameters in a long distance migrant bird of prey, the Swainson's Hawk (Buteo swainsoni), wintering in central Argentina. We analyzed serum concentration of urea, uric acid, cholesterol, and triglycerides and assessed age and sex related differences in these parameters. A body condition index was obtained from the resultant residuals of the regression of body mass and a morphometric measure. No statistical differences were observed among sex and age groups for urea, uric acid and triglyceride serum concentration. However, cholesterol concentration differed among male and female hawks, which could be related to the gain of body mass in wintering grounds at differential rates. The mean values of the four parameters were in the range of those recorded in the Common Buzzard (Buteo buteo), indicating good nutritional condition of the population we studied. Forearm length was the morphometric variable that better correlated with body mass. The resultant body condition index was only correlated with triglyceride concentration, suggesting that this index could be valuable in future work dealing with the assessment of body fat storage in wintering and breeding hawks, as well as in stopover points on the migratory route. PMID- 15123178 TI - Adaptive energetics in house mice, Mus musculus domesticus, from the island of Porto Santo (Madeira archipelago, North Atlantic). AB - The bioenergetic strategies of house mice (Mus musculus domesticus) from the island of Porto Santo were investigated and compared with those of mice from mainland Portugal. Energy obtained from food ingestion was 18.2% lower in Porto Santo mice than in mainland mice (1.53 vs. 1.87 kJ/g/day). The same pattern was observed for metabolisable energy intake, which was 19.2% lower in island specimens (0.87 vs. 1.08 kJ/g/day for mainland specimens). Apparent digestibility was similar in both groups of mice. Basal metabolic rate (BMR) of Porto Santo individuals was low (1.16 ml O(2)/g/h), representing only 56% of the predicted value, based on body mass, while mainland individuals exhibited a BMR closer to the expected value, corresponding to 87% of the predicted value (1.80 ml O(2)/g/h). Thermoregulatory abilities within the range of 10-28 degrees C ambient temperature did not differ between island and mainland mice. Results suggest an adaptation of Porto Santo mice to the environmental aridity of the island of Porto Santo, leading to a conservative energetic strategy. PMID- 15123179 TI - Force-velocity properties of two avian hindlimb muscles. AB - Recent work has provided measurements of power output in avian skeletal muscles during running and flying, but little is known about the contractile properties of avian skeletal muscle. We used an in situ preparation to characterize the force-velocity properties of two hind limb muscles, the lateral gastrocnemius (LG) and peroneus longus (PL), in Wild Turkeys (Meleagris gallopavo). A servomotor measured shortening velocity for at least six different loads over the plateau region of the length-tension curve. The Hill equation was fit to the data to determine maximum shortening velocity and peak instantaneous power. Maximum unloaded shortening velocity was 13.0+/-1.6 L s(-1) for the LG muscle and 14.8+/ 1.0 L s(-1) for the PL muscle (mean+/-S.E.M.). These velocities are within the range of values published for reptilian and mammalian muscles. Values recorded for maximum isometric force per cross-sectional area, 271+/-28 kPa for the LG and 257+/-30.5 kPa for the PL, and peak instantaneous power output, 341.7+/-36.4 W kg(-1) for the LG and 319.4+/-42.5 W kg(-1) for the PL, were also within the range of published values for vertebrate muscle. The force-velocity properties of turkey LG and PL muscle do not reveal any extreme differences in the mechanical potential between avian and other vertebrate muscle. PMID- 15123180 TI - Ventilatory response to hyperoxia in the chick embryo. AB - In the avian embryo at term we measured the ventilatory response to hyperoxia, which lowers the chemoreceptor activity, to test the hypothesis that the peripheral chemoreceptors are tonically functional. Measurements of pulmonary ventilation (VE) were conducted in chicken embryos during the external pipping phase, at 38 degrees C, during air and hyperoxia, and during hypercapnia in air or in hyperoxia. Hyperoxia (95% O2) maintained for 30 min lowered VE by 15-20%, largely because of a reduction in breathing frequency (f). The oxygen consumption and carbon dioxide production of the embryo were not altered. The hyperoxic drop of VE was more marked in those embryos, which had higher values of normoxic VE. Hypercapnia, whether 2 or 5% CO2, increased VE, almost exclusively because of the increase in tidal volume (VT). The increase in VT was less pronounced when hypercapnia was associated with hyperoxia, and f slightly decreased. Hence, in hyperoxia, the VE response to CO2 was less than in air. The results are in support of the hypothesis that in the avian embryo, after the onset of breathing, the peripheral chemoreceptors exert a tonic facilitatory input on . This differs from neonatal mammals, where the chemoreceptors have minimal or no activity at birth, presumably because the increased arterial oxygenation with the onset of air breathing is a much more sudden phenomenon in mammals than it is in birds. PMID- 15123181 TI - Biochemical acclimation of metabolic enzymes in response to lowered temperature in tadpoles of Limnodynastes peronii. AB - We measured the rate at which the metabolic enzymes lactate dehydrogenase (LDH), citrate synthase (CS), and cytochrome c oxidase (CCO) acclimate in the response to lowered temperature in the axial muscle of tadpoles of Limnodynastes peronii (Anura: Myobatrachidae) over 6 weeks. In addition, we measured growth rates of the tadpoles kept at both temperatures and examined the activities of these enzymes in the liver tissue of the control group and cold-acclimated group at the end of the experiment. We found that LDH acclimates in axial muscle; the differences between the control and cold-acclimated group became apparent after 21 days. After 42 days, the activity of LDH in axial muscle in the cold acclimated group was 30% greater than the control group. Growth rates were maintained at 0.7 mm/week within both treatments despite the 10 degrees C difference in temperature between experimental groups. Both LDH and CS were increased in activity in the liver (5 and 1.3 times greater, respectively, in the cold-acclimated group). The thermal sensitivity (Q(10)) of LDH was between 20 and 30 degrees C in the cold-acclimated group (1.2+/-0.01) when compared to the control group (1.6+/-0.15). The rate at which acclimation in this species occurs is appropriate for seasonal changes in temperature, and these animals may not be able to respond to a rapid drop in temperature. PMID- 15123182 TI - Adenosine triphosphate levels in steelhead (Oncorhynchus mykiss) eggs: an examination of turnover, localization and role. AB - The dynamics of energy production and utilization in fish eggs before and shortly after fertilization may be critical for embryo survival. Therefore, the current study examined the turnover of adenosine triphosphate (ATP) as well as examined the possible role and localization of ATP in unfertilized steelhead (Oncorhynchus mykiss) eggs and early embryos. The mean ATP level in unfertilized steelhead eggs was 1.92+/-0.10 (mean+/-S.E.M., n=17) nmol ATP per egg. Exposure of the unfertilized egg to 10 degrees C water (water activation) and fertilization resulted in comparable and substantial decreases (approx. 20-50%) in egg ATP levels within 3 min. This suggests that the energy expended at fertilization is used in response to water activation rather than fertilization per se. Unfertilized eggs maintained in ovarian fluid for 9 days at 10 degrees C under air showed a progressive decline of fertility that reached zero after 6 days. In contrast, no significant changes were seen in ATP levels throughout this 9 days period. Thus, fertility does not positively correlate with egg ATP levels in stored eggs. In the unfertilized egg, the ATP stored in the yolk accounted for approximately 1.5% of the total egg ATP. After fertilization, the concentration of ATP in the yolk increased approximately seven-fold, with the yolk and blastoderm each now accounting for approximately 20% of the total remaining ATP. Finally, to estimate the changes in oxidative metabolism following fertilization, the cyanide (KCN)-sensitive decline in total ATP was determined for unfertilized eggs and 1 day embryos. In the presence of KCN, ATP levels declined to approximately 50% within 24 h in both unfertilized eggs as well as embryos; the rates of ATP decline were not different. Therefore, there was not a discernible increase in ATP generation by oxidative phosphorylation at the time of fertilization. PMID- 15123183 TI - Tissue osmolarity of the New Zealand land flatworm (Arthurdendyus triangulatus). AB - Tissue fluid osmolarity of flatworms kept with moist bark was 243+/-4 S.E.M. mOsm kg(-1). Tissue fluid osmolarity of those kept with water-saturated tissue paper was 205+/-5 S.E.M. mOsm kg(-1). Flatworms placed in water of 300 and 400 mOsm kg( 1) lost weight. Those placed in water of 0, 100 and 200 mOsm kg(-1) gained weight. This suggests that body tissue fluids were approximately 260 mOsm kg(-1). Tissue fluids were slightly hyperosmotic in external media of 200, 300 and 400 mOsm kg(-1), and strongly hyperosmotic at 0 and 100 mOsm kg(-1). The highest measured value of tissue osmolarity was 457 mOsm kg(-1) from a specimen in a medium of 400 mOsm kg(-1). The lowest value was 145 mOsm kg(-1) from a specimen in pure water. Transverse sections of flatworms from different media concentrations suggest that fluids are absorbed into or removed from all tissues. PMID- 15123184 TI - Influence of weaning and effect of post weaning dietary zinc and copper on electrophysiological response to glucose, theophylline and 5-HT in piglet small intestinal mucosa. AB - This study aimed to examine how weaning and how dietary zinc and/or copper fed post weaning may affect the electrophysiological response to glucose and to chloride secretagogues in piglet small intestine in vitro. Study 1 included 54 piglets (six litters of nine piglets). One piglet from every litter was killed 1 day before weaning. The remaining 48 piglets were allocated at weaning (28 d) to four dietary zinc treatments and subsequently killed 1-2, 5-6 or 14-15 days after weaning. Study 2 included 48 piglets (six litters of eight piglets) allocated to four dietary treatments, consisting of high or low dietary zinc with or without high dietary copper. All piglets in study 2 were killed 5-7 days after weaning. The in vitro studies in Ussing chambers showed that weaning resulted in increased ileal glucose absorption as well as increased neuroendocrine-regulated (activated by 5-HT) and cAMP-dependent (activated by theophylline) chloride secretion. High zinc supplementation reduced the responses to 5-HT and theophylline. The study did not reveal any influence of copper on these parameters. It is concluded that the positive effect of zinc supplementation on diarrhoea in weaned piglets may be due to zinc reducing the intestinal mucosal susceptibility to secretagogues that activate chloride secretion. PMID- 15123185 TI - The brain-pituitary-gonad axis in male teleosts, with special emphasis on flatfish (Pleuronectiformes). AB - The key component regulating vertebrate puberty and sexual maturation is the endocrine system primarily effectuated along the brain-pituitary-gonad (BPG) axis. By far most investigations on the teleost BPG axis have been performed on salmonids, carps, catfish and eels. Accordingly, earlier reviews on the BPG axis in teleosts have focused on these species, and mainly on females (e.g. 'Fish Physiology, vol. IXA. Reproduction (1983) pp. 97'; 'Proceedings of the Fourth International Symposium on the Reproductive Physiology of Fish. FishSymp91, Sheffield, UK, 1991, pp. 2'; 'Curr. Top. Dev. Biol. 30 (1995) pp. 103'; 'Rev. Fish Biol. Fish. 7 (1997) pp. 173'; 'Proceedings of the Sixth International Symposium on the Reproductive Physiology of Fish. John Grieg A/S, Bergen, Norway, 2000, pp. 211'). However, in recent years new data have emerged on the BPG axis in flatfish, especially at the level of the brain and pituitary. The evolutionarily advanced flatfishes are important model species both from an evolutionary point of view and also because many are candidates for aquaculture. The scope of this paper is to review the present status on the male teleost BPG axis, with an emphasis on flatfish. In doing so, we will first discuss the present understanding of the individual constituents of the axis in the best studied teleost models, and thereafter discuss available data on flatfish. Of the three constituents of the BPG axis, we will focus especially on the pituitary and gonadotropins. In addition to reviewing recent information on flatfish, we present some entirely new information on the phylogeny and molecular structure of teleost gonadotropins. PMID- 15123186 TI - Lobster hepatopancreatic epithelial single cell suspensions as models for electrogenic sodium-proton exchange. AB - Sodium-proton antiporters, also called Na+/H+ exchangers (NHE), are vital transmembrane proteins involved in multiple cellular functions including transepithelial ion transport and Na+ homeostasis of cells throughout the biological kingdom. Na+/H+ exchange is accelerated by cytosolic acidification and also by osmotically induced cell shrinking, thereby promoting recovery of the physiological pHi and volume. Eight isoforms of Na+/H+ exchangers have been cloned and characterized to date and share the same overall structure, but exhibit differences with respect to cellular localization, kinetic variables and plasma membrane targeting, in polarized epithelial cells. The electrogenic Na+ absorption across tight epithelia from invertebrates follow significantly different principles from the electroneutral Na+/H+ antiporter found in vertebrates. In all invertebrate cells examined, the antiporter displayed a 2Na+/1H+ transport stoichiometry and this transport was markedly inhibited by exogenous calcium and zinc. Na+/H+ exchangers (NHE) are present in crustacean hepatopancreatic cell type suspensions and are believed to function in acid-base regulation by driving the extrusion of protons across the hepatopancreatic epithelium in exchange for Na+ in the sea water. A brief review of current knowledge about Na+/H+ exchangers has been presented. In addition, understanding of hepatopancreatic Na+/H+ exchange is described as obtained after isolation of purified E-, R-, F- and B-cell suspensions from the whole organ by centrifugal elutriation. PMID- 15123187 TI - Seasonal temperature compensation in the horse mussel, Modiolus modiolus: metabolic enzymes, oxidative stress and heat shock proteins. AB - Seasonal collections of the subtidal horse mussel, Modiolus modiolus, from a depth of 10 m were made at the Isles of Shoals, New Hampshire to assess changes in overall energetic demand, measured as respiration, the maximal activities of rate-limiting enzymes of intermediate metabolism, level of oxidative stress, and the expression of heat shock proteins (HSP). Weighted respiration rates of mussels from winter collections were significantly lower than summer rates but decreased by less than 20%. Specific activities of several rate-limiting enzymes were measured in mussels from the summer and winter collections at the temperature of collection and the reciprocal seasonal temperature (15 and 5 degrees C). Comparisons of these enzyme activities and the protein concentrations of hexokinase and citrate synthase show that a quantitative strategy is used to acclimatize to winter temperatures by these rate-limiting enzymes of intermediate metabolism. The activities and protein concentrations of the antioxidant enzyme, Cu/Zn superoxide dismutase (SOD) is seasonally indistinguishable while the concentration of HSP 70 was greater in winter than in summer samples. These results show that mussels seasonally compensate for decreases in temperature by increasing the concentration of rate-limiting metabolic enzymes while maintaining the same level of antioxidant protection in summer and winter consistent with high aerobic metabolism in both winter and summer. Lastly, the significantly greater concentrations of HSP70 in winter samples suggests that protein chaperone functions must be maintained while other seasonal adjustments to cold temperatures are occurring. PMID- 15123188 TI - A comparative echocardiographic assessment of ventricular function in five species of sharks. AB - A comparative echocardiographic study was carried out on five shark species that differ in heart morphology and in aspects of their behavior and natural history. The study contrasted the ventricular function in the highly active mako shark (heart type IV) and four other sharks (heart type III) that differ in activity levels (i.e. the sedentary horn and swell sharks vs. the moderately active blue and smooth-hound sharks). All five species exhibited biphasic ventricular filling characterized by an early (conduit) and late (atrial systole) phase. In the mako shark, early filling was dominant as indicated by a higher early flow peak velocity, a greater early:late velocity ratio, and a greater early velocity time integral. In contrast, the late filling phase was the more important filling agent in the other species. Indices of systolic function such as ventricular ejection fraction and ventricular fractional shortening also reflect a more efficient cardiac pumping capacity in mako shark relative to the other four sharks. The comparative echocardiographic assessment of in vivo ventricular function integrates structural and functional features with shark activity level to arrive at a new perspective blending the occurrence of biphasic filling with functional concepts based on heart morphological typology and changing views regarding the role of factors such as central filling pressure and pericardial pressure on end-diastolic ventricular volume. PMID- 15123189 TI - Change in plasma cortisol and metabolites during the attendance period ashore in fasting lactating subantarctic fur seals. AB - Lactating fur seals (Arctocephalus tropicalis) alternate foraging trips at sea and pup attendance periods ashore. During the onshore nursing periods, lactating females do not have access to food and meet both their own metabolic requirements and milk production from their body reserve. Blood and milk samples were collected from females captured soon after their arrival ashore from a foraging trip and before their departure. Milk lipid but not milk protein content was positively related to the body condition index (BCI) of the female. During the 4 day attendance period ashore, females lost body mass, and plasma cortisol levels increased, whereas plasma urea concentration decreased and beta-hydroxybutyrate (beta-OHB) remained unchanged. The increase in cortisol level took place while blood urea concentration decreased and beta-OHB remained at a low level suggesting that it was independent from the transition from phase II to phase III that is indicative of the depletion of lipid body store as described in penguins. Thus, our results suggest that the increase in cortisol level in relation to decreasing BCI may either contribute to the mobilization of protein stores to ensure milk production when easily mobilized stores are used and/or could act as a re-feeding signal which is triggered well before females have depleted their body store. PMID- 15123190 TI - Post pressure hyperemia in the rat. AB - In prior studies in man, we have demonstrated that pressure-induced hyperemia lasts for prolonged periods as compared to the short-term hyperemia created by proximal arterial occlusion. We have analyzed this phenomenon in our well-studied rat model of skin blood flow. Skin blood flow was measured using laser Doppler techniques in Wistar Kyoto rats at the back, a nutritively perfused site, and at the plantar surface of the paw, where arteriovenous anastomotic perfusion dominates. A customized pressure feedback control device was used to vary applied pressures. At the back, pressures in excess of 80 mmHg resulted in occlusion, whereas at the paw 150 mmHg was required. The peak hyperemic flow after release of pressure was comparable to that elicited by proximal arterial occlusion with a blood pressure cuff. However, the post pressure hyperemia peak descended to a plateau value, which was 50-100% greater than baseline and continued for up to 20 min while the peak following proximal arterial occlusion returned to baseline within 4 min. At the back, post pressure hyperemia reached a maximum after application of 100 mmHg pressure. The application of higher pressures than required for occlusion produced no greater hyperemic response. At the paw, maximum post pressure hyperemia occurred at 100 mmHg, although this pressure level was not totally occlusive. Higher pressures resulted in no greater hyperemia. At the back, 10 min of occlusion produced a maximal peak value whereas 1 min was sufficient at the paw. The application of pressure to a heated probe with subsequent release, produced a hyperemic response. Normalized to baseline blood flow, there was no difference between the hyperemic responses at basal skin temperature and at 44 degrees C. There is a prolonged hyperemic response following local pressure occlusion compared to a much shorter period following proximal ischemic occlusion. One can presume two different mechanisms, one related to ischemia and the other a separate pressure related phenomenon. The thermal vasodilatory response is additive, not synergistic with the post pressure hyperemia we have demonstrated. This finding suggests that different mechanisms are involved in thermal vasodilation and post pressure hyperemia. PMID- 15123191 TI - Photoinhibition, bleaching susceptibility and mortality in two scleractinian corals, Platygyra ryukyuensis and Stylophora pistillata, in response to thermal and light stresses. AB - In the present study, we examined the effect of thermal stress on the photoinhibitory light threshold in a bleaching susceptible (Stylophora pistillata) and a bleaching resistant (Platygyra ryukyuensis) coral. Four light (0, 110, 520, 1015 micromol quantam(-2)s(-1)) and three temperature (26, 32 and 34 degrees C) conditions were used over a 3-h period, followed by 24- and 48-h recovery periods at approximately 21 degrees C under dim light. Dynamic photoinhibition could be detected in both P. ryukyuensis and S. pistillata under 520 and 1015 micromol quantam(-2)s(-1) at 26 degrees C and under 110 micromol quantam(-2)s(-1) at 32 degrees C only in S. pistillata. Chronic photoinhibition was recorded under 520 and 1015 micromol quantam(-2)s(-1) at 34 degrees C in P. ryukyuensis, and under 1015 micromol quantam(-2)s(-1) at 32 degrees C and under all light levels at 34 degrees C in S. pistillata. These results show that high temperature reduced the threshold light intensity for photoinhibition differently in two corals with different bleaching susceptibilities under thermal stress. No visual paling and mortality in P. ryukyuensis was observed at any treatment, even in chronically photoinhibited specimens, while paling and high mortality of S. pistillata was noted in all treatments, apart from samples at 26 degrees C. These observations suggest a potential role of the host in differential bleaching and mortality determination. PMID- 15123192 TI - Respiratory gas exchange in the desert flea Xenopsylla ramesis (Siphonaptera: Pulicidae): response to temperature and blood-feeding. AB - Xenopsylla ramesis is a flea species parasitizing gerbilline rodents in the deserts of the Middle East. This study was undertaken to determine metabolic requirements of the different developmental stages of the flea-life cycle as well as to investigate the metabolic response to temperature and starvation after blood feeding. A high resolution respirometry system was used to measure CO2 emission of fleas ranging in size from 0.166+/-0.006 mg (larvae) to 0.263+/-0.009 mg (adults). The free-living stages (larvae and adults) had significantly higher metabolic rates than the cocooned stages (pupae). CO2 emission rates of the larvae exceeded that of the adults by 2.6-fold and the pupae by 7.3 times. In the adults, both temperature and blood feeding significantly affected starvation level metabolism. Metabolism was temperature dependent with an average Q10 of 2.57 for females and 2.55 for males over the temperature range of 10-30 degrees C. No consistent decline in thermal sensitivity at higher ambient temperatures was evident. Fleas that had a blood meal prior to starvation had significantly higher metabolic rates (0. 86 +/- 0.008 x 10(-3) ml mg(-1) h(-1)) than fleas, which were newly emerged unfed adults (0.56 +/- 0.1 x 10(-3) ml mg(-1) h(-1)). Water content also differed between fed (range approx. 67-69% body mass) and newly emerged adults (range approx. 73-75% of body mass). Feeding may stimulate some as yet undetermined physiological process that causes differential metabolic response in starving, fed and unfed fleas. Characteristics of gas exchange in desert-dwelling fleas are reflective of the off-host life style in the protected microenvironment of the host nest or burrow, rather than as a response to any type of environmental extreme. PMID- 15123193 TI - Dietary fatty acid composition affects the repeat swimming performance of Atlantic salmon in seawater. AB - Repeated critical swimming performance trials (Ucrit) were performed on Atlantic salmon (Salmo salar) to test the null hypothesis that the source of dietary lipids (fish-based, poultry-based, and plant-based) does not influence exercise and recovery performance. Four diets were prepared by extensively replacing supplemental lipid from anchovy oil (AO; 100% AO at 150 g/kg) with cold pressed flaxseed oil (FO; 25% AO, 75% FO), sunflower oil (SO; 25% AO, 75% SO), or poultry fat (PF; 25% AO, 75% PF). These diets had equivalent protein and energy concentrations, but due to the different supplemental lipid sources, varied widely in their fatty acid composition. Fish fed AO had a significantly higher (P<0.05) first Ucrit (2.62+/-0.07 body lenght s(-1)) than those fed PF (2.22+/ 0.12 body lenght s(-1)) that had low muscle ratios of n-3 highly unsaturated fatty acids (n-3 HUFA) to saturated fatty acids (SFA) and arachidonic acid (AA), and high levels of oleic acid. Fish in the FO and SO diet groups swam as well as AO-fed fish in both swimming trials. The performance of fish fed AO decreased significantly (P<0.05) during the second swimming trial (i.e. Ucrit2/Ucrit1=0.92+/-0.02). No significant differences occurred between diet groups for the second swim trial. There was a positive correlation between both n 3 HUFA/SFA and n-3 HUFA/AA ratios, and Ucrit1. A negative correlation was found between dietary AA and oleic acids, and Ucrit1. The present study suggests that low dietary n-3 HUFA/ SFA and n-3 HUFA/AA ratios may negatively affect swimming performance. The former possibly can be offset by increasing linoleic acid in the presence of nutritionally adequate n-3 HUFA (e.g. SO diet). Lipid supplements consisting largely of vegetable oils did not compromise fish cardiorespiratory physiology under the conditions of this study. PMID- 15123194 TI - Comparative studies on the wide frequency band electrocardiogram and vectorcardiogram in pigeon and mouse. AB - Wide frequency band ECG and vectorcardiogram in anesthetized pigeon and mouse were studied from the standpoint of comparison. The key results were as follows: in pigeon, the direction of the main QRS was inverted in leads II, III and aVF, and upright in lead aVR, which was contrary to that in mouse. The T wave was upright in leads II, III and aVF, but inverted in lead aVR in pigeon, which was the same as that in mouse. In pigeon, there was a large notch on the upstroke of the S wave in lead II without exception, but there was no such notch in the corresponding lead in mouse. The QRS vector loop in the frontal plane lay between -90 and -180 degrees in pigeon, while that of mouse lay between 0 and 90 degrees. The relative power of high frequency range (80-1000 Hz) of the QRS in lead II was approximately 15% in pigeon, but 55% in mouse. The direction of the main QRS was contrary in pigeon and mouse because the subepicardial muscles were depolarized before the subendocardial muscles in pigeon, but the latter were depolarized before the former in mouse. The direction of the T waves was the same in both pigeon and mouse because subepicardial muscles were all repolarized before subendocardial muscles. PMID- 15123195 TI - Ion transport mechanisms in the mesonephric collecting duct system of the toad Bufo bufo: microelectrode recordings from isolated and perfused tubules. AB - It is not clear how and whether terrestrial amphibians handle NaCl transport in the distal nephron. Therefore, we studied ion transport in isolated perfused collecting tubules and ducts from toad, Bufo bufo, by means of microelectrodes. No qualitative difference in basolateral cell membrane potential (Vbl) was observed between tubules and ducts in response to ion substitutions, inhibitor and agonist applications. Cl- substitution experiments indicated a small Cl- conductance in the basolateral membrane. The apical membrane did not have a significant Cl- conductance. Luminal [Na+] steps and amiloride application showed a small apical Na+ conductance. Arginine vasotocin depolarized Vbl. The small apical Na+ conductance indicates that the collecting duct system contributes little to NaCl reabsorption when compared to aquatic amphibians. In contrast, Vbl rapidly depolarized upon lowering of [Na+] in the bath, demonstrating the presence of a Na+-coupled anion transporter. [HCO3-] steps revealed that this transporter is not a Na+-HCO3- cotransporter. Together, our results indicate that a major task of the collecting duct system in B. bufo is not conductive NaCl transport but rather K+ secretion, as shown by our previous studies. Moreover, our results indicate the presence of a novel basolateral Na+-coupled anion transporter, the identity of which remains to be elucidated. PMID- 15123196 TI - Cost of living in free-ranging degus (Octodon degus): seasonal dynamics of energy expenditure. AB - Animals process and allocate energy at different seasons at variable rates, depending on their breeding season and changes in environmental conditions and resulting physiological demands. Overall total energy expenditure, in turn, should either increase in some seasons due to special added demands (e.g. reproduction) or it could simply remain at about the same level, in which case the animals must show compensatory rebalancing of other expenditures that can be reduced. To test for the alternative hypotheses of seasonal variability or compensation, we measured total daily energy expenditure (DEE) in free-living degus (Octodon degus) at four seasons and followed this with determinations of basal metabolic rate (BMR) in the laboratory in the same individuals. DEE varied seasonally but was only significantly different (lower) in summer (non-breeding season), with a DEE:BMR ratio of only 1.6, whereas autumn, winter and spring DEE values were statistically indistinguishable from one another and showed DEE:BMR ratios ranging from 1.9 to 2.2. Our values of DEE in the field fall within the broad range of allometric expectation for herbivorous mammals in general, but the ratios of DEE:BMR are lower than expected. This, together with the lack of strong major shifts in total levels of DEE, suggests that degus are showing compensatory shifts among various categories of energy expenditure that allow them to manage their overall energy balance by minimizing total expenditure. PMID- 15123197 TI - Seasonal changes in angiotensin converting enzyme activity in male and female frogs (Rana esculenta). AB - Gonad, lung, kidney and serum angiotensin converting enzyme (ACE) activities were determined by specific substrate hydrolysis in male and female Rana esculenta over 1 year. Ovary ACE activity showed the highest values among the different tissues, with a significant peak (223+/-52 nmol min(-1) mg protein(-1)) in late winter-early spring. Testis ACE activity followed a significant seasonal cycle, increasing from September to peak in April (2.5+/-0.8 nmol min(-1) mg protein( 1)) and then decreased in the post-reproductive period. Lung and kidney ACE activities were not correlated with the annual reproductive cycle phases. In serum a peak of activity was present in the post-reproductive period both in male and female frogs. The present data show a correlation between ACE and the annual reproductive cycle of R. esculenta. PMID- 15123198 TI - Cortisol response of green sturgeon to acid-infusion stress. AB - Cortisol and lactate are classic indicators of stress in fishes and their interactive effects on metabolism during recovery from stress have recently become a subject of more intense study. We examined how stressing green sturgeon through acid infusion affected the cortisol response and lactate metabolism in green sturgeon (Acipenser medirostris). Both lactic acid (0.3 M) and HCl (0.3 N) infusion (infusion volumes 1.5 ml kg(-1)) elicited an immediate cortisol response (21.61+/-4.61 ng ml(-1) and 17.50+/-3.00 ng ml(-1), respectively). Lactic acid prolonged the cortisol response compared to HCl (90 min vs. 25 min). Neutralized lactate (0.23 M; with 1 N NaOH; final pH 7.8) and NaCl (0.9%) infusion (infusion volumes 1.5 ml kg(-1)) did not affect plasma cortisol. Sturgeon infused with lactic acid showed a faster rate of lactate disappearance from plasma than those with neutralized lactic acid. We relate these findings to lactate metabolism following exercise, acid-infusion and air immersion stress in fishes. PMID- 15123199 TI - Calcium regulation in crustaceans during the molt cycle: a review and update. AB - Epithelial cells of the gut, gills, antennal glands and integument regulate calcium concentrations in crustaceans during the molt cycle. A cellular calcium transport model has been proposed suggesting the presence of calcium pumps, cation antiporters and calcium channels in transporting epithelial membranes that regulate the movements of this cation across the cell layer. Basolateral calcium transport during postmolt appears mainly regulated by the low affinity NCX antiporter, while calcium regulating 'housekeeping' activities of these cells in intermolt are controlled by the high affinity calcium ATPase (PMCA). A model is proposed for the involvement of the epithelial ER in the massive transepithelial calcium fluxes that occur during premolt and postmolt. This model involves the endoplasmic reticulum SERCA and RyR proteins and proposed cytoplasmic unstirred layers adjacent to apical and basolateral plasma membranes where calcium activities may largely exceed those in the bulk cytoplasmic phase. A result of the proposed transepithelial calcium transport model is that large quantities of calcium can be moved through these cells by these processes without affecting the low, and carefully controlled, bulk cytoplasmic calcium activities. PMID- 15123200 TI - Effect of hypotonic shock on cultured pavement cells from freshwater or seawater rainbow trout gills. AB - The effect of hypotonic shock on cultured pavement gill cells from freshwater (FW)- and seawater (SW)-adapted trout was investigated. Exposure to 2/3rd strength Ringer solution produced an increase in cell volume followed by a slow regulatory volume decrease (RVD). The hypotonic challenge also induced a biphasic increase in cytosolic Ca(2+) with an initial peak followed by a sustained plateau. Absence of external Ca(2+) did not modify cell volume under isotonic conditions, but inhibited RVD after hypotonic shock. [Ca(2+)](i) response to hypotonicity was also partially inhibited in Ca-free bathing solutions. Similar results were obtained whether using cultured gill cells prepared from FW or SW fishes. When comparing freshly isolated cells with cultured gill cells, a similar Ca(2+) signalling response to hypotonic shock was observed regardless of the presence or absence of Ca(2+) in the solution. In conclusion, gill pavement cells in primary culture are able to regulate cell volume after a cell swelling and express a RVD response associated with an intracellular calcium increase. A similar response to a hypotonic shock was recorded for cultured gill cells collected from FW and SW trout. Finally, we showed that calcium responses were physiologically relevant as comparable results were observed with freshly isolated cells exposed to hypoosmotic shock. PMID- 15123201 TI - Role of urea in the postprandial urine concentration cycle of the insectivorous bat Antrozous pallidus. AB - Insectivorous bats, which feed once daily, produce maximally concentrated urine only after feeding. The role of urea as an osmolyte in this process was investigated in pallid bats (Antrozous pallidus) in the laboratory. Following a 24-h fast, plasma and urine were sampled before and 2 h after feeding in postprandial (PP) animals and before and 2 h after similar treatment without feeding in nonfed (NF) animals. Food consumption by PP animals and handling of NF animals had no effect on blood water content as measured by hematocrit and plasma oncotic pressure. Food consumption increased both plasma osmolality (P(osm)) and plasma urea (P(urea)) by as much as 15%. Food consumption also increased urine osmolality (U(osm)) and urine urea (U(urea)) by 50-100%. Feeding increased U(osm) regardless of changes in P(osm), and elevation of U(osm) resulted primarily from increased U(urea). In NF bats, P(osm) and P(urea) were unchanged, while U(osm) and U(urea) increased by as much as 25%. Again, increased U(osm) resulted primarily from increased U(urea). The PP urine concentration cycle of pallid bats resulted from increased urea excretion in response to apparent rapid urea synthesis. Bats rapidly metabolized protein and excreted urea following feeding when body water was most plentiful. PMID- 15123202 TI - Lipopolysaccharide evokes microaggregation reactions in hemocytes isolated from tobacco hornworms, Manduca sexta. AB - Insect cellular immune reactions to bacterial infection include nodule formation. Eicosanoids mediate several cellular actions in the nodulation process, including formation of hemocyte microaggregates, an early step. In previous work, we reported that isolated hemocytes produce and secrete eicosanoids that influence hemocyte behavior in response to bacterial challenge. We also reported that microaggregate formation in response to challenge was mediated by prostaglandins (PGs), but not by products of the lipoxygenase (LOX) pathways. In this paper we describe experiments designed to test the idea that exposing isolated hemocytes to lipopolysaccharide (LPS) evokes formation of hemocyte microaggregates and this cellular action is mediated by PGs. Results show that isolated hemocyte preparations challenged with LPS formed more hemocyte microaggregates than unchallenged preparations (6.9x10(3) microaggregates/ml hemolymph vs. 2.5x10(3) microaggregates/ml hemolymph). LPS challenge stimulated formation of hemocyte microaggregates in a dose dependent manner. Experimental groups pretreated with cyclooxygenase inhibitors produced fewer hemocyte microaggregates in response to LPS challenge than untreated control groups. The formation of hemocyte microaggregates was not influenced by LOX inhibitors. Furthermore, the influence of dexamethasone was reversed by supplementing the experimental groups with the eicosanoid precursor fatty acid molecule, arachidonic acid and PGH(2). Palmitic acid, which is not substrate for eicosanoid biosynthesis, did not reverse the effects of dexamethasone on the formation of microaggregates. The LOX product 5(S)hydroperoxyeicosa-6E,8Z,11Z,14Z-tetraenoic acid also did not reverse the effects of dexamethasone. These results are consistent with similar investigations performed with bacterial suspensions. We infer that isolated hemocyte preparations recognize and react to LPS by forming microaggregates and this reaction is mediated by PGs, but not products of the LOX pathway. PMID- 15123203 TI - Seasonal and daily changes in the capacity for nonshivering thermogenesis in the golden hamsters housed under semi-natural conditions. AB - Nonshivering thermogenesis (NST) is a main source of heat for many small mammals. It undergoes seasonal changes, being the highest in winter and the lowest in summer. Such acclimatization can ensure winter survival for species living in moderate or cold climates. Nevertheless, not only seasonal, but also daily changes in the capacity for NST seem to be of great importance. In this study, the effects of season and time of day on the temperature of brown adipose tissue (T(BAT)), preferred ambient temperature (PT(a)) and activity after noradrenaline (NA) injections in golden hamsters (Mesocricetus auratus) housed under semi natural conditions were investigated. Animals were kept in outdoor enclosures and experienced natural changes in both, photoperiod and ambient temperature (T(a)). NA-induced hyperthermia was the largest during autumn (mean increase in T(BAT) by 0.74+/-0.04 degrees C), while during summer increase in T(BAT) was similar to that recorded in control (saline-injected) animals (0.16+/-0.05 degrees C and 0.24+/-0.04 degrees C, respectively). In spring hyperthermia was intermediary (0.57+/-0.05 degrees C). Daily variations in the response to NA depended on the season. In summer, the largest increase in T(BAT) (0.45+/-0.1 degrees C) was recorded during the first part of the day, while in autumn-in the middle of the day and night (1.1+/-0.1 degrees C and 0.9+/-0.1 degrees C, respectively). In spring, all NA injections induced large increase in T(BAT) except for the injection in the middle of the night. The largest decrease in PT(a) after NA administration was recorded in autumn (mean decrease by 1.5+/-0.3 degrees C). Both, seasonal and daily changes in the capacity for NST reflect different demands for heat dependently on the time of the year and time of the day. It can be concluded that although long history of breeding in captivity, golden hamsters preserved ability to survive in natural environment. PMID- 15123204 TI - Age- and sex-dependent changes in pulse pressure in fowl aorta. AB - Chickens (males more than females) have higher blood pressure (BP) than most mammals and spontaneously develop vascular neointimal plaques (NP) and diffuse subendothelial thickening in the lower segment of the abdominal aorta (AbA, referred to as 'NP-prone area') that partly resemble atherosclerotic lesions in mammals. NP areas, which are larger in males, have a causal relationship with incremental increases in BP during maturation. We hypothesize that decreased wall distensibility and altered hemodynamic forces at the NP-prone area may contribute to the NP formation. We measured pressure pulse wave (PW) and systolic and diastolic BP along the descending aorta in anesthetized chickens at different ages using an intravascular microtip transducer and calculated pulse pressure (PP) as an indicator for artery distensibility. At all ages examined and in both sexes, the PW showed a sharper peak at the more peripheral locations and the amplitude of the PW increased as it descended the aorta. PP, expressed as relative increases from the PP in the aortic arch (%), was 40.4+/-12.6 and 71.4+/ 18.6 at the AbA and ischiadic artery, respectively, in young males (24-27 weeks); 23.5+/-8.6 and 43.8+/-16.2 in adults (72-75 weeks); and 5.4+/-3.4 and 9.1+/-4.9 in chicks (5-7 weeks). Location-dependent increases in PP were significantly higher in young males (P<0.05). The PP increases in females were not different among the three age groups. The contour of the PW in the proximal aorta changes in older birds, exhibiting steeper increases in the ascending and descending limbs, suggesting that faster wave reflection from the periphery augments peak systolic pressure. NP was most frequently seen in the lower segment of the abdominal aorta in older males. These results suggest that: (1) site-dependent increases in PP amplitude are marked in young males, possibly reflecting a reduction in arterial wall elasticity enhanced by incremental rises in BP, and (2) NP formation may contribute to the stiffness of aortic walls in the NP-prone area. PMID- 15123205 TI - Differential tolerance of body fluid dilution in two species of tropical hermit crabs: not due to osmotic/ionic regulation. AB - The tropical intertidal hermit crabs Clibanarius taeniatus and Clibanarius virescens were examined for differences in survival and physiological responses in low salinity. We found that C. taeniatus survived better in dilute seawater than C. virescens and that these species did not differ in their abilities to regulate haemolymph osmolarity, ionic concentration of the haemolymph or body fluid volume. We also found no difference in oxygen consumption between the species when acutely exposed to a range of temperature and salinity combinations. It is concluded that the greater survival in dilute seawater by C. taeniatus compared to C. virescens is due to a greater tolerance of dilution of body fluids by C. taeniatus. Differences in tolerance to dilute seawater may influence the habitat preferences of these species within the same geographical area. PMID- 15123206 TI - The development of homeothermy in mink (Mustela vison). AB - In mink (Mustela vison) kits newborn mortality is very high. One of the major causes of death is hypothermia. The objectives of this study were to observe the development of thermoregulation in mink kits, and their ability to maintain their body temperature during the postnatal period (1-50 days of age). Based on the kit's body weight (BW), and rectal and ambient temperature measurements during cold (+4 degrees C) and warm (+40 degrees C) exposures, a homeothermy index (HI) and cooling and warming rates were calculated. No significant differences in the body temperatures were found between the kits and the dam after 36 days of age. The kits were able to maintain homeothermy by 22 days of age (HI 90%). The body cooling rate was 0.88+/-0.04 degrees C min(-1) on day 1 but only 0.35+/-0.03 degrees C min(-1) at 22 days of age. The body WR was lower: day 1, 0.85+/-0.04 degrees C min(-1) and 0.22+/-0.03 degrees C min(-1) at 22 days of age. All measured and calculated thermophysiological variables were significantly influenced by BW and age of the kit. PMID- 15123207 TI - Sulfamethazine advances puberty in male chicks by effecting a rapid increase in gonadotropins. AB - A sulfonamide, sulfamethazine (SMZ) has been shown to have a robust, progonadal effect. The mechanism of action of SMZ, however, is unknown. Our hypothesis is that the compound may act centrally and/or at the level of the pituitary. Four experiments were completed to test that hypothesis. Chicks exposed to a continuous photoperiod and fed a diet containing 0.2% SMZ showed an exponential increase in testes size. When 6 weeks of age (5 weeks on the SMZ diet), experimentals had testes weight nine times heavier than controls. Profiles for thyroid and gonadotropin plasma hormones suggested that T(3) was transiently lower in experimentals solely during the first week on treatment, while thyroxine levels were not different from controls. In contrast, luteinizing hormone (LH) and follicle stimulating hormone (FSH) were significantly elevated at the initial 1-week sampling point and remained elevated throughout the entire experiment. In a follow-up study, LH was found significantly higher than controls by 48 h after initially consuming the compound. When T(3) was added to the SMZ diet at 0.5 ppm, the progonadal effect of SMZ was attenuated. Importantly, chronic intake of T(3) delayed but did not block the stimulatory effect of SMZ for increasing plasma LH. We conclude that since one of the primary effects of SMZ is to increase rapidly plasma gonadotropins, data suggest the compound is acting at the level of the brain or pituitary to stimulate early gonadal development in chicks. PMID- 15123208 TI - Saturated fatty acids suppress adrenocorticotropic hormone (ACTH) release from rat anterior pituitary cells in vitro. AB - We studied whether fatty acids modify adrenocorticotropic hormone (ACTH) release induced by stimulation with corticotropin-releasing hormone (CRH) from rat anterior pituitary cells. Stimulation with CRH (0.01-100 nmol/l) significantly and concentration-dependently increased ACTH release, which was synergistically enhanced by the simultaneous stimulation with 1 nmol/l arginine-vasopressin. Addition of saturated fatty acids (butyrate, caprylate, laurate, palmitate and stearate) in a medium at 1 mmol/l, despite effects on the basal release, significantly reduced the ACTH release induced by CRH (1 nmol/l) stimulation. Caprylate suppressed ACTH release in a concentration-dependent manner. However, unsaturated C18 and C20 fatty acids (oleate, linolate, linolenate and arachidonate) at 1 mmol/l significantly increased the basal release, but none of them suppressed CRH (1 nmol/l)-induced ACTH release. In the presence of caprylate (1 mmol/l), CRH (1 nmol/l)-stimulated increase in cellular calcium ion concentration was diminished. From these results we conclude that saturated fatty acids have a suppressing effect on CRH-induced ACTH increase in primary cultured rat anterior pituitary cells. PMID- 15123209 TI - Chemical composition of the sponge Hymeniacidon sanguinea from the Canary Islands. AB - The fatty acid composition of the lipids from the sponge Hymeniacidon sanguinea was investigated and 73 acids were identified. Three of them were new and their structures, elucidated by gas chromatography/mass spectrometry, were identified as 13-methyl-icosanoic, 15-methyl-docosanoic and 3,13-dimethyl-tetradecanoic. Only 12 sterols were present, with cholestanol predominating in the volatile fraction; 22 compounds were identified, mainly hydrocarbons and fatty acids. In the n-butanol fraction, we found mainly nitrogen-containing compounds. The compositions of sterols and lipids in the sponge collected from two locations with different ecological conditions (Canary Islands and Black Sea) were compared. PMID- 15123210 TI - Characterization of bovine serum ferritin-binding proteins. AB - Ferritin-binding proteins (FBPs) in bovine serum were characterized by ferritin immunoassay, ferritin-binding activity, and immunoblotting. Serum ferritin, but not tissue ferritin, was precipitated by centrifugation at 14000 x g for 30 min, and bovine spleen ferritin added to bovine serum was precipitated by centrifugation at 1650 x g for 20 min. Two FBPs (FBP1 and FBP2) were purified from bovine serum by sequential chromatography on bovine spleen ferritin Sepharose 4B affinity and Sephacryl S-300 columns. FBP1 separated into 82 kDa- and 26 kDa-bands on SDS-PAGE, while FBP2 separated into 55 kDa- and 26 kDa-bands. FBP1 and FBP2 were identified as IgM and IgG, respectively, by immunoblotting with alkaline phosphatase-labeled antibodies specific for bovine IgM, IgG, and IgA heavy chains. Given these results, we suggest that bovine FBPs are autoantibodies (IgM and IgG) to ferritin and that circulating ferritin exists as an immune complex. PMID- 15123211 TI - Hepatopancreas gluconeogenesis and glycogen content during fasting in crabs previously maintained on a high-protein or carbohydrate-rich diet. AB - The present study assessed the effect of different fasting times on the in vitro gluconeogenic capacity of Chasmagnathus granulata crabs previously adapted to a high-protein (HP) or carbohydrate-rich (HC) diet using the incorporation of [U (14)C]l-lactate or [U-(14)C]l-alanine into glucose. We also recorded haemolymphatic glucose and hepatopancreatic glycogen levels. In the HP group, on the third day of fasting there were decreases in the synthesis of glucose from (14)C-alanine and in haemolymph glucose. After 15 days of fasting, haemolymph glucose and hepatopancreatic glycogen levels were maintained by an increase in the conversion of (14)C-alanine into glucose. However, after 21 days of fasting the gluconeogenic capacity was decreased and hepatopancreas glycogen concentration was reduced. In the HC group, hepatopancreatic glycogen was the energy source during the first 6 days of fasting. Gluconeogenesis from (14)C lactate decreased after 6 days of fasting, remaining low until 21 days of fasting. The conversion of (14)C-alanine into glucose was increased after 15 days fasting and hepatopancreatic glycogen was raised in relation to that present after a 6-day fasting. In both dietary groups the stabilization in the levels of haemolymph glucose after 21 days fasting may result from a reduction in metabolic rate during restricted feeding. PMID- 15123212 TI - Physiological and biochemical responses to dietary protein in the omnivore passerine Zonotrichia capensis (Emberizidae). AB - We studied the physiological, biochemical and morphological responses of the omnivore sparrow Zonotrichia capensis, a small opportunistic passerine from Central Chile acclimated to high- and low-protein diets. After 4 weeks of acclimation to 30% (high-protein group) or 7% (low-protein group) dietary casein, we collected urine and plasma for nitrogen waste production and osmometry analysis, and measured gross renal morphology. Plasma osmolality and hematocrit were not significantly affected by dietary treatment, but urine osmolality was higher in the high-protein group than in the low-protein group. Kidney and heart masses were higher in animals acclimated to the high-protein diet. Mean total nitrogen waste was significantly higher in the high-protein group, but the proportions of nitrogen excreted as uric acid, urea and ammonia were unaffected by diet. Our data suggest that the response of Z. capensis to an increase in dietary protein content is through greater amounts of total nitrogen excretion and hypertrophy of kidney structures, without any modification of the proportion of excretory compounds. PMID- 15123213 TI - Electrophysiological demonstration of independent olfactory receptor types and associated neuronal responses in the trout olfactory bulb. AB - The present study attempts to highlight the principles by which peripheral olfactory information of across- and within-class odorant signals is transformed into bulbar neuron responses. For this purpose, we performed electro-olfactogram cross-adaptation and mixture experiments as well as single unit recording of olfactory bulb neurons using amino acid, bile acid and F-prostaglandin stimulants in brown and rainbow trout. The results show that amino acids, a bile acid and a F-prostaglandin activate independent receptor types. However, within the class of amino acids, different receptor types are only partially independent. Neurons responsive to bile acid and amino acids were segregated to the mid-dorsal and latero-posterior olfactory bulb, respectively. Of the 43 responsive olfactory bulb neurons studied in brown trout, 41 showed specificity for one odorant class. Olfactory bulb neurons gained responsiveness to new amino acids with increasing stimulant concentration. We conclude that different odorant classes activate specific neurons located in different regions of the trout olfactory bulb, and that information distinguishing related amino acids can be represented in a limited number of bulbar neurons with distinct response profiles under the conditions investigated. PMID- 15123215 TI - Differential energy costs of winter acclimatized common spiny mice Acomys cahirinus from two adjacent habitats. AB - The common spiny mouse Acomys cahirinus, of Ethiopian origin, has a widespread distribution across arid, semi-arid and Mediterranean parts of the Arabian sub region. We compared the daily energy expenditure (DEE), water turnover (WTO) and sustained metabolic scope (SusMS=DEE/resting metabolic rate) of two adjacent populations during the winter. Mice were captured from North- and South- facing slopes (NFS and SFS) of the same valley, comprising mesic and xeric habitats, respectively. Both DEE and SusMS winter values were greater in NFS than SFS mice and were significantly greater than values previously measured in the summer for these two populations in the same environments. However, WTO values were consistent with previously established values and were not significantly different from allometric predictions for desert eutherians. We suggest that physiological plasticity in energy expenditure, which exists both temporally and spatially, combined with stable WTO, perhaps reflecting a xeric ancestry, has enabled A. cahirinus to invade a wide range of habitats. PMID- 15123214 TI - Mechanisms contributing to intracellular pH homeostasis in an immortalised human chondrocyte cell line. AB - The maintenance of chondrocyte pH is an important parameter controlling cartilage matrix turnover rates. Previous studies have shown that, to varying degrees, chondrocytes rely on Na(+)/H(+) exchange to regulate pH. HCO(3)(-)-dependent buffering and HCO(3)(-)-dependent acid-extrusion systems seem to play relatively minor roles. This situation may reflect minimal carbonic anhydrase activity in cartilage cells. In the present study, the pH regulation of the human chondrocyte cell line, C-20/A4 has been characterised. Intracellular pH (pH(i)) was measured using the H(+)-sensitive fluoroprobe BCECF. In solutions lacking HCO(3)(-)/CO(2), pH(i) was approximately 7.5, and the recovery from intracellular acidification was predominantly mediated by a Na(+)-dependent, amiloride- and HOE 694-sensitive process. A small additional component which was sensitive to chloro-7-nitrobenz-2 oxa-1,3-diazole, an inhibitor of the V-type H(+)-ATPase, was also apparent. In solutions containing HCO(3)(-)/CO(2), pH(i) was approximately 7.2. Comparison of buffering capacity in the two conditions showed that this variable was not significantly augmented in HCO(3)(-)/CO(2)-containing media. The recovery from intracellular acidification was more rapid in the presence of HCO(3)(-)/CO(2), although under these conditions it was again largely dependent on Na(+) ions and inhibited by amiloride and HOE 694. A small component was inhibited by SITS, although this effect did not reach the level of statistical significance. These findings indicate that HCO(3)(-)-dependent processes play only a minimal role in pH regulation in C-20/A4 chondrocytes. pH regulation instead relies heavily on the Na(+)/H(+) exchanger together with a H(+)-ATPase. The absence of extrinsic (HCO(3)(-)/CO(2)) buffering is likely to reflect the low levels of carbonic anhydrase in these cells. In addition to providing fundamental information about a widely-used cell line, these findings support the contention that the unusual nature of pH regulation in chondrocytes reflects the paucity of carbonic anhydrase activity in these cells. PMID- 15123216 TI - Respiratory response to temperature and hypoxia in the zebra mussel Dreissena polymorpha. AB - The effects of temperature acclimation, acute temperature variation and progressive hypoxia on oxygen consumption rates (VO2) were determined for the zebra mussel Dreissena polymorpha. In the first experiment, after acclimation to 5, 15 or 25 degrees C for at least 2 weeks, VO2 was determined at 5 degrees C increments from 5 to 45 degrees C. VO2 increased in all three acclimation groups from 5 to 30 degrees C, corresponding to the normal ambient temperature range for this species. Mussels displayed imperfect temperature compensation at temperatures above 15 degrees C, but exhibited little acclimatory ability below 15 degrees C. In the hypoxia experiment, VO2 was determined over the course of progressive hypoxia, from full saturation (oxygen tension [PO2]=160 Torr [21.3 kPa]) to a PO2 at which oxygen uptake ceased (<10 Torr [1.3 kPa]). Mussels were acclimated to either 5, 15 or 25 degrees C for at least 2 weeks and their respiratory response to progressive hypoxia was measured at three test temperatures (5, 15 and 25 degrees C). The degree of oxygen regulation increased with increasing test temperature, particularly from 5 to 15 degrees C, but decreased with increasing acclimation temperature. The decreased metabolic rate observed for warm-acclimated animals, particularly in the upper portion of the temperature range of the zebra mussel, may allow for conservation of organic energy stores during warm summer months. Compared to other freshwater bivalves, D. polymorpha is a relatively poor oxygen regulator, corresponding with its preference for well-oxygenated aquatic habitats. In addition, a new quantitative method for determining the degree of oxygen regulation is presented. PMID- 15123217 TI - Skeletal muscle bioenergetics: a comparative study of mitochondria isolated from pigeon pectoralis, rat soleus, rat biceps brachii, pig biceps femoris and human quadriceps. AB - The metabolism of mitochondria isolated from five functionally different skeletal muscles is compared. Data for a single ectothermic preparation are also reported. The mitochondria were prepared in yields of 44+/-7% from 50 to 100 mg muscle. The muscle content of mitochondrial protein ranged between 2 and 40 g kg(-1). Twelve specific activities of key enzymes and metabolic systems were determined, 10 of these in functional assays with respiratory measurements. The specific activities of glutamate dehydrogenase, alpha-glycerophosphate dehydrogenase, and exo-NADH oxidase differed considerably among muscle sources. Seven specific activities, including very central reactions, showed low among-muscle variation. The activity of ATP synthesis, for instance, was 1.0-1.3 mmol min(-1) g(-1) mitochondrial protein, 25 degrees C. In vitro data were extrapolated to in vivo conditions of the muscles. The calculated rates of respiration and ATP synthesis were in accordance with reported tissue activities. Pigeon pectoralis mitochondria showed a unique cytochrome spectrum and a respiratory chain activity that might effect simultaneous carbohydrate and fatty acid respiration. In mitochondria from the other muscles, the respiratory chain activity balanced the carbohydrate oxidation capacity. In all muscles, the respiratory capacity exceeds that needed for oxidative phosphorylation. This may secure maximal mitochondrial ATP synthesis during maximal work rates and high cellular [Ca(2+)]. PMID- 15123218 TI - The NHLBI COPD clinical research network. PMID- 15123219 TI - Elevation of retinyl ester level in the lungs of rats following repeated intraperitoneal injections of retinoic acid or retinoyl glucuronide. AB - Retinoic acid (RA) is involved in the development of both the conducting airway and alveolar portions of the lung. RA plays a key role in the induction of the formation of alveolar septa. Retinoyl beta-glucuronide (RAG), an endogenous retinoid, acts like RA, but is much less cytotoxic. We examined the effect of daily intraperitoneal doses of RA and RAG (1.66 micromol/kg) in dimethyl sulfoxide (DMSO) and cotton seed oil on the stores of retinol and retinyl ester (RE) in the lung of rats. In two separate experiments, one involving normal rats, and the other involving elastase-treated rats, there was up to a 9-fold increase in REs content in the lungs of rats that received RA or RAG for 12 days as compared to Control rats. The accumulation was most profound when RA was injected in DMSO and least when RAG was injected in cotton seed oil. The reason for the accumulation of REs in the lung is not clearly known. PMID- 15123220 TI - Onset of action of formoterol/budesonide in single inhaler vs. formoterol in patients with COPD. AB - Formoterol is a beta(2)-agonist bronchodilator that combines a fast onset of action with a long duration of broncholytic effect. An increasing documentation is showing that the combination of a long acting beta(2)-adrenoceptor agonist bronchodilator and an inhaled corticosteroid targets the airways obstruction in patients with COPD. In this study, we have explored whether the acute addition of an inhaled corticosteroid influences the fast bronchodilator response to formoterol. A total of 20 patients with stable COPD were randomized. Single doses of formoterol/budesonide 2 x (4.5/160)microg or formoterol 2 x 4.5 microg were given via Turbuhaler. Serial measurements of FEV(1) were performed over 60 min. Formoterol/budesonide elicited a significantly larger mean FEV(1)-AUC(0-15 min) than formoterol alone. Also the change in FEV(1) 15 min after inhalation of formoterol/budesonide combination (0.197 l; 95% CI: to 0.142-0.252) was greater than that induced by formoterol alone (0.147 l; 95% CI: to 0.092-0.201). The mean increases in FEV(1) were always higher after budesonide/formoterol than formoterol alone, although both treatments induced a significant improvement over baseline at each explored time point. Even the FEV(1)-AUC(0-60 min) after formoterol/budesonide was significantly larger than that after formoterol. Both treatments induced a significant reduction in VAS score but did not modify heart rate in a statistically significant manner. This study indicates that the addition of budesonide influences the fast onset of action of formoterol, but does not induce systemic effects, in patients with stable COPD. PMID- 15123221 TI - Effect of salbutamol on smoking related cough. AB - Smokers have an increased prevalence of chronic cough and may complain of exacerbation of cough when attempting smoking cessation. We investigated the use of smokers cough as a model for testing anti-tussive agents. The effect of salbutamol was compared with placebo in healthy adult smokers. In a randomised double blind crossover study the effect of 400 microg salbutamol via MDI plus spacer versus placebo was studied. Cough was assessed before and after the first cigarette of the day (received at 20 minutes) and throughout the day. Cough frequency, citric acid cough challenge, change in cough symptoms and peak flow were recorded. Salbutamol reduced the mean cough frequency between 0 and 20 min. A mean of 4.5 compared to 6 on placebo (p<0.05). A significant reduction in cough followed cigarette consumption in those on placebo. Mean pre-cigarette 6 compared to 3.9 post-cigarette (p<0.02). The citric acid concentration causing two coughs (C2) at 60 min increased on salbutamol. Geometric mean 278.8 compared to 190.4 mM on placebo (p<0.03). Cough frequency is reduced in smokers following a cigarette. The reduction in cough frequency and evoked cough after salbutamol suggests that beta agonists have modest activity in smoking related cough and that smokers cough represents a sensitive model to test anti-tussive activity. PMID- 15123222 TI - The role of neutrophils in LPS-induced changes in pulmonary function in conscious rats. AB - We have previously reported on a model of lipopolysaccharide (LPS)-induced pulmonary inflammation in rats, where LPS-challenged animals develop a significant pulmonary neutrophilia and mucus hypersecretion. In the current studies, we utilized whole body plethysmography and computer assisted data acquisition to examine changes in pulmonary parameters, e.g. frequency (f) tidal volume and Penh as a measure of bronchoconstriction, due to LPS-challenge in conscious rats. Compared to saline challenge, LPS-challenged rats displayed a significant increase in (f) which began within 30 min, peaked by 2 h and remained elevated up to 24 h. Mirroring this increase in (f) was a decrease in the observed tidal volume of LPS-challenged rats. Additionally, compared to saline challenge, LPS-challenge provoked a significant and spontaneous bronchoconstriction, as measured by Penh, 2 h after challenge. In order to further understand these observed LPS-induced pulmonary changes, we utilized two classes of pulmonary obstructive disease standards, namely, bronchodilators and anti-inflammatory agents, and examined their ability to affect the spontaneous bronchoconstriction and the increase in (f) seen at two discrete time points, i.e. 2 and 24 h after LPS-challenge. While ineffective on either the 2 h increase in (f) or the LPS-induced inflammation, animals pretreated with salbutamol (10 mg/kg, p.o.) were protected from the increase in (f) seen at the 24 h time point after LPS-challenge. In contrast, when animals were pretreated with theophylline (10 mg/kg, p.o.) no effect on the LPS-induced pulmonary inflammation or increase in (f) was noted. Meanwhile, in animals pretreated with either betamethasone (3 mg/kg, p.o.) or SB207499 (10 mg/kg, p.o.), a PDE4 inhibitor, doses previously shown to block the LPS-induced neutrophilic inflammation, the persistent increase in (f) seen at 24 h was attenuated, but neither compound was able to attenuate either the increase in (f) or the spontaneous bronchoconstriction seen at 2 h. In summary, the intra-tracheal LPS-challenge of rats results in pulmonary inflammation and dysfunction, which is similar to that seen in COPD patients. We conclude that the early increase in (f) and bronchoconstriction are not dependent upon airway inflammation, but airway inflammation most likely contributes to the persistent increase in (f) seen at 24 h. PMID- 15123223 TI - Salmeterol/fluticasone propionate in a Single Inhaler Device versus theophylline+fluticasone propionate in patients with COPD. AB - STUDY OBJECTIVES: The aim of this study was to compare the relative efficacy in terms of improvement in symptoms and lung function of salmeterol/fluticasone propionate (SLM/FP) combination administered through the Diskus inhaler versus theophylline (THEO) added to FP Diskus in patients with stable chronic obstructive pulmonary disease (COPD). METHODS AND MEASUREMENTS: Eighty patients were randomized to receive 4 months of treatment in one of two treatment groups: (1) fixed combination of SLM 50 microg and FP 500 microg Diskus, 1 inhalation twice daily; or (2) FP Diskus 500 microg, 1 inhalation twice daily, plus oral titrated THEO twice daily. Patients attended the clinic before and after 4, 8, 12 and 16 weeks of treatment for evaluations of pulmonary function, and dyspnea, which was assessed using an analogic visual scale. Also the supplemental salbutamol use was measured. RESULTS: . Sixty-six patients completed the 4-month treatment period: 37 patients receiving SLM/FP and 29 patients receiving THEO+FP. Patients were withdrawn for various reasons, the most common of which were poor compliance with the protocol, exacerbation and GI events. A gradual increase in FEV(1) was observed with each treatment. Maximum significant increases in FEV(1) over baseline values that were observed after 4 months of treatment were as follows: SLM/FP 0.172 l (95% CI: 0.084-0.260) and THEO+FP 0.155 l (95% CI: 0.054 0.256). SLM/FP experienced significantly (p<0.05) greater improvements in dyspnea, and required significantly fewer supplemental salbutamol treatments than the THEO+FP group. CONCLUSIONS: Our results suggest that SLM/FP combination may provide substantial benefits in both physiologic and clinical outcomes in symptomatic patients with COPD. It also causes a more effective control than THEO+FP. PMID- 15123224 TI - One week treatment with salmeterol does not prevent early and late asthmatic responses and sputum eosinophilia induced by allergen challenge in asthmatics. AB - Salmeterol is an effective long-acting beta(2)-agonist bronchodilator, able to inhibit, as a single dose, asthmatic responses induced by several stimuli including allergen, and the subsequent increase in sputum eosinophilia. Aim of the present study was to investigate whether these effects of salmeterol persisted after 1 week of continuous treatment, or whether a loss of the bronchoprotective effects of salmeterol can occur over time. We investigated in a cross-over double blind placebo-controlled study, the protective effect of 1 week treatment with salmeterol on allergen-induced early and late responses and the associated airway inflammation in 15 atopic asthmatic subjects. Eosinophil percentage and Eosinophil Cationic Protein (ECP) concentration in peripheral blood and in hypertonic saline induced sputum were measured at baseline and 24 h after allergen inhalation. Salmeterol partially inhibited early asthmatic response, but it did not inhibit late asthmatic response in comparison with placebo. Salmeterol did not inhibit also the increase in sputum eosinophils percentage 24 h after allergen inhalation (E%, median: 22.7 and 15%, after placebo and after salmeterol respectively, p=n.s. between two post-allergen sputum samples). Also, the increase in blood eosinophils and both sputum and serum ECP at 24 h after allergen challenge was not affected by salmeterol pre treatment. In conclusion, 1 week treatment with salmeterol causes a loss of its protective effect on allergen-induced airway bronchoconstriction, and does not prevent the subsequent increase in sputum and serum eosinophilic markers. PMID- 15123225 TI - Effect of inhaled N-acetylcysteine on hydrogen peroxide exhalation in healthy subjects. AB - N-acetylcysteine (NAC) has antioxidant properties and its oral administration decreased H(2)O(2) exhalation in patients with chronic obstructive pulmonary disease. In this study we tested whether inhaled NAC could suppress H(2)O(2) levels in exhaled breath condensate (EBC) of eight healthy subjects that have never smoked (never-smokers). Original NAC solution (ACC vial, 300 mg NAC in 3 ml solvent), NAC-placebo (vehicle), sterile 0.9% NaCl or distilled water were nebulized via the pneumatic De Vilbiss nebulizer once daily every 7 days and H(2)O(2) and thiols exhalation was measured just before, 30 min and 3 h after the end of drug administration. Additional in vitro experiments were performed to evaluate NAC stability during nebulization, reactivity with H(2)O(2) and possible H(2)O(2) generation in aqueous NAC solutions. NAC almost completely abolished H(2)O(2) exhalation 30 min after inhalation (0.02+/-0.04 vs. 0.21+/-0.09 microM, p<0.001). However, 3 h later the H(2)O(2) levels raised 1.8-fold from baseline (p<0.01). Other inhaled solutions did not affect H(2)O(2) levels. Mean thiol concentration in EBC rose (p<0.05) after treatment with NAC and reached 1.03+/ 0.48 microM at 3 h. Although, 25 and 50 mM NAC completely inhibited H(2)O(2) peroxidase-luminol-dependent chemiluminescence, detectable amounts of H(2)O(2) were generated in NAC solutions. It was accompanied by moderate loss of -SH groups. Catalase and ascorbic acid prevented H(2)O(2) formation in NAC solutions. In conclusion inhaled NAC revealed biphasic effect on H(2)O(2) exhalation in healthy subjects, which depends on direct H(2)O(2) scavenging and H(2)O(2) generation related to drug oxidation. The net result of these processes may determine anti- or pro-oxidant action of inhaled NAC. PMID- 15123226 TI - Allergen challenge alters lymphocyte phosphodiesterase activity in horses with heaves. AB - Heaves is an allergic airway disease in horses characterised by reversible airway obstruction, bronchial hyperresponsiveness and airway inflammation associated with a Th(2) response. Cyclic nucleotide-dependent signalling pathways can regulate lymphocyte function. In this study, we examined lymphocyte PDE activity comparing horses with heaves to healthy control animals. Total PDE activity and the effects of isoenzyme selective inhibitors were measured before, 5 and 24 h after the start of a 7 h allergen challenge. Allergen challenge had no effect on either total cAMP PDE activity or its inhibition by the PDE4 selective inhibitor, rolipram, and the non-selective PDE inhibitor, theophylline. In contrast, the PDE3 selective inhibitor, quazinone, caused significantly greater inhibition of cAMP PDE activity before challenge in the heaves susceptible group. Additionally, total cGMP PDE activity was significantly lower 24 h after the start of challenge in the heaves affected group (11+/-2 and 21+/-3 pmol/min/mg for heaves and control animals, respectively) and the PDE5 selective inhibitor, zaprinast, caused significantly less inhibition in the heaves group at this time point. The functional significance of these findings was explored by examining the effect of PDE3, PDE4 and PDE5 selective inhibitors on mitogen-induced mononuclear cell proliferation before and 24 h after the start of allergen challenge. Proliferation decreased after challenge in the heaves group (stimulation index=328+/-110 and 200+/-72 before and after challenge, respectively) whilst remaining constant in the control group (stimulation index=161+/-13 and 183+/-45 before and after challenge, respectively). However, all three PDE inhibitors caused a similar amount of inhibition at each time point and the effect of a combination of a PDE3 and a PDE5 inhibitor was simply additive in both groups. These results suggest differences in the control of lymphocyte PDE activity in horses with heaves. PMID- 15123227 TI - Hyperresponsiveness to tobacco dust extract in sensitized guinea pig trachea. AB - The role of pre-existing airway inflammation in the pathogenesis of occupational airway disease is poorly understood. Previously we studied an extract of tobacco dust (TDE) and determined that it causes concentration dependent contractions of nonsensitized guinea pig trachea (GPT). In the present study animals were sensitized using Ovalbumin (OA) and subsequently challenged with an aerosol of 2.5% OA on day 21. A control group of nonsensitized GPs were divided into rings in which the epithelium was retained (EPI+) or removed (EPI-). Concentration related contractions of sensitized and nonsensitized GPTs were elicited with TDE. Sensitized GPTs demonstrated a greater contractile response to TDE than did nonsensitized GPTs. In nonsensitized animals the EPI- GPTs demonstrated a lesser response to TDE than did the EPI+. Similar findings were demonstrated in sensitized GPTs with and without epithelium. When epithelium was removed, sensitized and nonsensitized GPTs behaved similarly. Moreover, sensitized GPTs without epithelium and nonsensitized with epithelium responded similarly. These findings suggest that presensitization with an unrelated antigen enhances the response to an occupational agent and that in sensitized animals at least part of the enhanced response is mediated by the epithelial layer. PMID- 15123228 TI - Beta2-integrin adhesion caused by eotaxin but not IL-5 is blocked by PDE-4 inhibition and beta2-adrenoceptor activation in human eosinophils. AB - We investigated the effect and mechanism(s) of PDE-4 treatment with concurrent beta2-adrenoceptor stimulation on human eosinophil adhesion mediated by beta2 integrin in vitro. Eosinophils were pretreated with either rolipram, a PDE-4 inhibitor, alone or combined with salmeterol, a beta2-adrenoceptor agonist, before activation with either eotaxin or IL-5. Beta2-integrin mediated adhesion was assessed as adherence to BSA, an established surrogate for ICAM-1. Rolipram caused progressive blockade (77.7 +/- 6.2%) of adhesion elicited by eotaxin. Maximal blockade of IL-5-activated adhesion by rolipram was substantially less (29.9 +/- 5.2%). Salmeterol + rolipram synergistically enhanced the blockade of eotaxin-activated adhesion. Eotaxin also caused approximately 50% increase in surface CD11b expression, which was blocked additively by rolipram + salmeterol. By contrast, CD11b upregulation caused by IL-5 was not blocked by rolipram + salmeterol. Rolipram also attenuated cPLA2 phosphorylation caused by eotaxin but did not block IL-5-induced phosphorylation. We conclude that rolipram blocks expression of CD11b and inhibits cPLA2 phosphorylation in human eosinophils, thus blocking eotaxin-induced adhesion of beta2-integrin. IL-5-induced adhesion likely utilizes a different upstream mechanism in regulation of integrin adhesion. PMID- 15123229 TI - Cholinergic responsiveness of the individual airway after allergen instillation in sensitised pigs. AB - Allergen exposure of sensitised lungs produces bronchial hyperresponsiveness in vivo associated with airway inflammation and remodelling. It is unclear if hyperresponsiveness is also present in airways in vitro under similar conditions of drug provocation as carried out in vivo, and at different times after allergen challenge. This study records responsiveness of individual airway segments to acetylcholine (ACh) in sensitised bronchi after instillation of allergen (ovalbumin, OA). Airway histology and sensitivity and maximum effects to ACh were recorded 1, 24 and 72 h and 1 week after OA. OA-instilled airways exhibited eosinophilia and epithelial proliferation. Physiological recordings showed no change in maximum contractions of airway segments to acetylcholine placed in the airway lumen except at 24 h where they were reduced. In contrast maximum contractions to ACh to the airway adventitia were reduced at all times except 1 week, with the greatest change occurring at 24 h. There were no changes in airway sensitivity to either route of ACh in OA-instilled airways but the difference in sensitivity to adventitial and lumenal ACh was reduced. Results show that allergen does not produce hyperresponsiveness at the airway wall but it may alter an interaction between airway smooth muscle and other structural components of the airway. PMID- 15123230 TI - Budesonide/formoterol in a single inhaler rapidly relieves methacholine-induced moderate-to-severe bronchoconstriction. AB - Inhalers containing corticosteroids and long-acting beta2-agonists are becoming increasingly important in asthma management. A rapid effect is important to patients, particularly during exacerbations. We compared the onset of bronchodilation and patient-perceived relief from dyspnoea following single inhaler budesonide/formoterol or salmeterol/fluticasone in a model of acute bronchoconstriction. A randomised, double-blind, double-dummy, single-dose, crossover study included 27 outpatients with asthma (mean age 35 years; mean FEV1 90% predicted normal). Immediately following methacholine-induced bronchoconstriction (fall in FEV1 > or = 30%), patients inhaled budesonide/formoterol (160/4.5 microg, 1 or 2 inhalations; Symbicort Turbuhaler), salmeterol/fluticasone (50/250 microg; Seretide Diskus) or placebo on 4 study days. Lung function and Borg score were assessed for 30 min. During methacholine induced provocation (final mean FEV1 62.5% of baseline), mean Borg score increased 10-fold (from 0.3 to 3.0 units). Hereafter, mean FEV1 at 3 min improved significantly more after budesonide/formoterol 1 and 2 inhalations (37 and 38%, respectively) than after salmeterol/fluticasone (23%; P < 0.001) or placebo (10%; P < 0.001). Median recovery times to 85% of baseline FEV1 were shorter for budesonide/formoterol (1 or 2 inhalations: 3.3 and 2.8 min, respectively) than salmeterol/fluticasone (8.9 min; P < 0.001) and placebo (> 30 min). One min after budesonide/formoterol, dyspnoea was significantly reduced (Borg score -0.86 units, both doses) compared with salmeterol/fluticasone (-0.55 units; P < 0.05) and placebo (-0.23 units; P < 0.001). Budesonide/formoterol provides immediate bronchodilation, faster than salmeterol/fluticasone, which patients can feel during acute methacholine-induced bronchoconstriction. PMID- 15123231 TI - Proteinase-activated receptor (PAR)-mediated vasorelaxation in pulmonary arteries from normotensive and hypoxic pulmonary hypertensive rats. AB - Proteinase-activated receptor (PAR)-mediated vasorelaxant responses were examined in main pulmonary artery preparations from control rats and rats exposed to hypoxia (10% oxygen) for 1 or 4 weeks to induce pulmonary hypertension. Trypsin and the PAR-2 peptide, SLIGRL, relaxed phenylephrine precontracted preparations, with maximum responses the same as the maximum response to acetylcholine. Responses to trypsin and SLIGRL were abolished by endothelium removal or a nitric oxide (NO) synthase inhibitor, and were, therefore, due to release of endothelium derived NO. In pulmonary arteries from rats exposed to hypoxia for 1 week, the potencies and maximal responses for acetylcholine and trypsin were markedly reduced compared with data in control rats, but these values were restored to normal in arteries from 4-week hypoxic rats. In contrast, the potency of SLIGRL was unchanged in arteries from either group of hypoxic rats. The data for trypsin and acetylcholine are consistent with previous findings for a variety of endothelium-dependent vasodilators and reflect an impairment of endothelial function in early hypoxic pulmonary hypertension. The data for SLIGRL emphasise that one cannot necessarily predict changes in PAR-2-mediated endothelium dependent vasorelaxation in disease states from data with the classical endothelium-dependent vasodilator, acetylcholine. PMID- 15123232 TI - Effective attenuation of endotoxin-induced acute lung injury by 2,3 diacetyloxybenzoic acid in two independent animal models. AB - The pathology of acute lung injury (ALI) is often modeled in animal studies by the administration of lipopolysaccharide (LPS), which results in an endotoxemia with sequelae similar to that seen in acute respiratory distress syndrome (ARDS). Here we report the results of two studies designed to examine the efficacy of a novel agent, 2,3-diacetyloxybenzoic acid (2,3-DABA), in the treatment of LPS induced ALI. In two separate animal models, 2,3-DABA was effective in significantly reducing lung microvascular permeability, a condition commonly seen in ARDS, which results in pulmonary edema and respiratory insufficiency. In each model, it is demonstrated that the mechanism by which 2,3-DABA exerts this effect occurs subsequent to the recruitment of neutrophils to the site of inflammation. Lung permeability was significantly decreased in both models by treatment with 2,3-DABA, suggesting that this agent, either alone or in combination therapy, may be useful in the treatment of ALI associated with ARDS. PMID- 15123233 TI - Breaking down tumor defenses. PMID- 15123234 TI - Nano-tailoring; stitching alterations on viral coats. PMID- 15123235 TI - Heparin-induced cancer cell death. PMID- 15123236 TI - PDZ domains and their ligands. PMID- 15123237 TI - Enhanced intravenous transgene expression in mouse lung using cyclic-head cationic lipids. AB - Herein, we report enhanced intravenous mouse lung transfection using novel cyclic head-group analogs of usually open-head cationic transfection lipids. Design and synthesis of the new cyclic-head lipid N,N-di-n-tetradecyl-3,4-dihydroxy pyrrolidinium chloride (lipid 1) and its higher alkyl-chain analogs (lipids 2-4) and relative in vitro and in vivo gene transfer efficacies of cyclic-head lipids 1-4 to their corresponding open-head analogs [lipid 5, namely N,N-di-n-tetradecyl N,N-(2-hydroxyethyl)ammonium chloride and its higher alkyl-chain analogs, lipids 6-8] have been described. In stark contrast to comparable in vitro transfection efficacies of both the cyclic- and open-head lipids, lipids 1-4 with cyclic heads were found to be significantly more efficient (by 5- to 11-fold) in transfecting mouse lung than their corresponding open-head analogs (5-8) upon intravenous administration. The cyclic-head lipid 3 with di-stearyl hydrophobic tail was found to be the most promising for future applications. PMID- 15123238 TI - The first fluorescent diboronic acid sensor specific for hepatocellular carcinoma cells expressing sialyl Lewis X. AB - Carbohydrate antigens with subterminal fucosylation have been implicated in the development and progression of several cancers, including hepatocellular carcinoma (HCC). Fluorescent sensors targeting fucosylated carbohydrate antigens could potentially be used for diagnostic and other applications. We have designed and synthesized a series of 26 diboronic acid compounds as potential fluorescent sensors for such carbohydrates. Among these compounds, 7q was able to fluorescently label cells expressing high levels of sLex (HEPG2) within a concentration range of 0.5 to 10 microM. This compound (7q) did not label cells expressing Lewis Y (HEP3B), nor cells without fucosylated antigens (COS7). This represents the first example of a fluorescent compound labeling cells based on cell surface carbohydrate structures. PMID- 15123239 TI - An improved method for the synthesis of cellulose membrane-bound peptides with free C termini is useful for PDZ domain binding studies. AB - SPOT synthesis permits parallel synthesis and screening of thousands of cellulose membrane-bound peptides to study protein-protein interactions in a proteomic context. Recognition of C-terminal residues is one of the most common binding features of PDZ domains. Unfortunately, most solid support-bound peptide libraries lack a free C terminus due to C-terminal fixation on the solid support. To overcome this restriction, we developed a robust methodology based on our previous strategy for generating peptides with authentic C termini. To validate this improved method, we screened a human peptide library of 6223 C termini with the syntrophin PDZ domain. Furthermore, using the same library, new peptide ligands derived from membrane proteins and receptors were found for the ERBIN PDZ domain. Finally, we identified the protein kinase breakpoint cluster region, which is known as a negative regulator of cell proliferation and oncogenic transformation, as an ERBIN ligand. PMID- 15123240 TI - Context-dependent behavior of the enterocin iterative polyketide synthase; a new model for ketoreduction. AB - Heterologous expression and mutagenesis of the enterocin type II polyketide synthase (PKS) system suggest for the first time that the association of an extended set of proteins and substrates is needed for the effective production of the enterocin-wailupemycin polyketides. In the absence of its endogenous ketoreductase (KR) EncD in either the enterocin producer "Streptomyces maritimus" or the engineered host S. lividans K4-114, the enterocin minimal PKS is unable to produce benzoate-primed polyketides, even when complemented with the homologous actinorhodin KR ActIII or with EncD active site mutants. These data suggest that the enterocin PKS requires EncD to serve a catalytic and not just a structural role in the functional PKS enzyme complex. This strongly implies that EncD reduces the polyketide chain during elongation rather than after its complete assembly, as suggested for most type II PKSs. PMID- 15123241 TI - Targeting specific PDZ domains of PSD-95; structural basis for enhanced affinity and enzymatic stability of a cyclic peptide. AB - A cyclic peptide, Tyr-Lys-c[-Lys-Thr-Glu(betaAla)-]-Val, incorporating a beta-Ala lactam side chain linker and designed to target the PDZ domains of the postsynaptic density protein 95 (PSD-95), has been synthesized and structurally characterized by NMR while free and bound to the PDZ1 domain of PSD-95. While bound, the lactam linker of the peptide makes a number of unique contacts outside the canonical PDZ binding motif, providing a novel target for PDZ-domain specificity as well as producing a 10-fold enhancement in binding affinity. Additionally, the cyclization greatly enhances the enzymatic stability, increasing the duration that the peptide inhibits the association between PSD-95 and glutamate receptors, effectively inhibiting the clustering of kainate receptors for over 14 hr after application. Highly specific regulation of kainate receptor action may provide a novel route for treatment of drug addiction and epilepsy. PMID- 15123242 TI - Novel real-time sensors to quantitatively assess in vivo inositol 1,4,5 trisphosphate production in intact cells. AB - Real-time observation of messenger molecules in individual intact cells is essential for physiological studies of signaling mechanisms. We have developed a novel inositol 1,4,5-trisphosphate (IP(3)) sensor based on the pleckstrin homology (PH) domain from phospholipase C (PLC) delta. The environmentally sensitive fluorophore 6-bromoacetyl-2-dimethyl-aminonaphtalene was conjugated to the genetically introduced cysteine at the mouth of the IP(3) binding pocket for enhanced IP(3) selectivity and for rapid and direct visualization of intracellular IP(3) > or = 0.5 microM as fluorescence emission decreased. The probe, tagged with arginine-rich sequences for efficient translocation into various cell types, revealed a major contribution of Ca2+ influx to PLC-mediated IP(3) production that boosts Ca2+ release from endoplasmic reticulum. Thus, our IP(3) probe was extremely effective to quantitatively assess real-time physiological IP(3) production via those pathways formed only in the intact cellular configuration. PMID- 15123243 TI - Poly(beta-amino ester)s promote cellular uptake of heparin and cancer cell death. AB - Heparin/heparan sulfate-like glycosaminoglycans (HSGAGs) are involved in diverse cellular processes in the extracellular matrix (ECM). The biological effect of HSGAGs depends on disaccharide content and physiological location within the ECM. HSGAGs are also brought into cells during membrane transcytosis and growth factor signaling while protein bound. We sought to probe the impact of free HSGAGs within the cell by using heparin as a model HSGAG. A library of poly(beta-amino ester)s, which internalize DNA, was examined for the capacity of its members to internalize heparin. Fourteen polymers enabled heparin internalization. The most efficacious polymer reduced murine melanoma cell growth by 73%. No glycosaminoglycan was as efficacious as highly sulfated, full-length heparin. Internalized heparin likely interferes with transcription factor function and subsequently induces apoptotic cell death. Therefore, internalized heparin is a novel mechanism for inducing apoptosis of cancer cells. PMID- 15123244 TI - ADP-specific sensors enable universal assay of protein kinase activity. AB - Two molecular sensors that specifically recognize ADP in a background of over 100 fold molar excess of ATP are described. These sensors are nucleic-acid based and comprise a general method for monitoring protein kinase activity. The ADP-aptamer scintillation proximity assay is configured in a single-step, homogeneous format while the allosteric ribozyme (RiboReporter) sensor generates a fluorescent signal upon ADP-dependent ribozyme self-cleavage. Both systems perform well when configured for high-throughput screening and have been used to rediscover a known protein kinase inhibitor in a high-throughput screening format. PMID- 15123245 TI - Ligand-mediated transcription elongation control using triplex-based padlock oligonucleotides. AB - Triplex-forming oligonucleotides (TFOs) provide useful tools for the artificial regulation of gene expression at the transcriptional level. They can become topologically linked to their DNA target upon circularization, thereby forming very stable triple helical structures. These "padlock oligonucleotides" are able to interfere with transcription elongation when their target site is located in the transcribed region of a gene. In vitro transcription experiments showed that a bacterial RNA polymerase was stopped at the site of triple-helix formation, whereas expression of a reporter gene was inhibited in live cells. In both cases, the padlock oligonucleotide was more efficient at inhibiting transcription elongation than a linear TFO, and the inhibition was observed only in the presence of a triplex stabilizing agent. These results provide new insights into the ligand-modulated locking of padlock oligonucleotides around their DNA target. PMID- 15123246 TI - Simultaneous characterization of the reductive unfolding pathways of RNase B isoforms by top-down mass spectrometry. AB - A novel method for characterization of the simultaneous reductive unfolding pathways of five isoforms of bovine pancreatic ribonuclease B (RNase B) is demonstrated. The results indicate that each isoform unfolds reductively through two three-disulfide-containing structured intermediates before proceeding to the fully reduced form, as in the reductive unfolding pathways of the A variant lacking the carbohydrate chain. The rates of reduction of bovine pancreatic ribonuclease A (RNase A) and RNase B and the formation and consumption of their reductive intermediates are identical, indicating that the unfolding events necessary to expose disulfide bonds for reduction are not affected by the oligosaccharide. The method utilizes top-down mass spectrometry and a naturally occurring tag on the protein, viz. the carbohydrate moiety, to obtain unfolding information of an ensemble of protein isoforms and is a generally applicable methodological advance for conducting folding studies on mixtures of different proteins. PMID- 15123247 TI - Structural determinants of CDK4 inhibition and design of selective ATP competitive inhibitors. AB - A number of selective inhibitors of the CDK4/cyclin D1 complex have been reported recently. Due to the absence of an experimental CDK4 structure, the ligand and protein determinants contributing to CDK4 selectivity are poorly understood at present. Here, we report the use of computational methods to elucidate the characteristics of selectivity and to derive the structural basis for specific, high-affinity binding of inhibitors to the CDK4 active site. From these data, the hypothesis emerged that appropriate incorporation of an ionizable function into a CDK2 inhibitor results in more favorable binding to CDK4. This knowledge was applied to the design of compounds in the otherwise CDK2-selective 2-anilino-4 (thiazol-5-yl)pyrimidine pharmacophore that are potent and highly selective ATP antagonists of CDK4/cyclin D1. The findings of this study also have significant implications in the design of CDK4 mimic structures based on CDK2. PMID- 15123248 TI - Profiling enzyme activities in vivo using click chemistry methods. AB - Methods for profiling the activity of enzymes in vivo are needed to understand the role that these proteins and their endogenous regulators play in physiological and pathological processes. Recently, we introduced a tag-free strategy for activity-based protein profiling (ABPP) that utilizes the copper(I) catalyzed azide-alkyne cycloaddition reaction ("click chemistry") to analyze the functional state of enzymes in living cells and organisms. Here, we report a detailed characterization of the reaction parameters that affect click chemistry based ABPP and identify conditions that maximize the speed, sensitivity, and bioorthogonality of this approach. Using these optimized conditions, we compare the enzyme activity profiles of living and homogenized breast cancer cells, resulting in the identification of several enzymes that are labeled by activity based probes in situ but not in vitro. PMID- 15123249 TI - Generation of new landomycins by combinatorial biosynthetic manipulation of the LndGT4 gene of the landomycin E cluster in S. globisporus. AB - A 3 kb DNA fragment from the Streptomyces globisporus 1912 landomycin E (LaE) biosynthetic gene cluster (lnd) was completely sequenced. Three open reading frames were identified, lndGT4, lndZ4, and lndZ5, whose probable translation products resemble a glycosyltransferase, a reductase, and a hydroxylase, respectively. Studies of generated mutants from disruption and complementation experiments involving the lndGT4 gene allowed us to determine that LndGT4 controls the terminal L-rhodinose sugar attachment during LaE biosynthesis and that LndZ4/LndZ5 are responsible for the unique C11-hydroxylation of the landomycins. Generation of the novel landomycins F, G, and H in the course of these studies provided evidence for the flexibility of lnd glycosyltransferases toward their acceptor substrates and a basis for initial structure-activity relationships within the landomycin family of antibiotics. PMID- 15123250 TI - Synthesis, characterization, and cytotoxicity of a series of estrogen-tethered platinum(IV) complexes. AB - Several estrogen-tethered platinum(IV) complexes were prepared and characterized by ESI-MS and (1)H NMR spectroscopy. Their design was inspired by the observation that estrogen receptor-positive cells exposed to the hormone are sensitized to cisplatin. Intracellular reduction of bis-estrogen-cis diamminedichloroplatinum(IV), BEP(n) (where n = 1-5 methylene groups between Pt and estrogen), occurs to afford cisplatin and two equivalents of the linker modified estrogen. The ability of BEP(n) to induce overexpression of HMGB1 was established by immunofluorescence microscopy. The cytotoxicity of the compounds was evaluated in ER(+) MCF-7 and ER(-) HCC-1937 human breast cancer cell lines. BEP3 selectively induces overexpression of HMGB1 in MCF-7 cells, compared to HCC 1937 cells, and enhances their sensitivity (IC(50) = 2.1 +/- 0.4 microM versus 3.7 +/- 0.9 microM, respectively) to the compound. The difference in compound activities and the potential of compounds of this class for treating breast and ovarian cancer are discussed. PMID- 15123251 TI - A bacterial acetyltransferase capable of regioselective N-acetylation of antibiotics and histones. AB - The Salmonella enterica chromosomally encoded AAC(6')-Iy has been shown to confer broad aminoglycoside resistance in strains in which the structural gene is expressed. The three-dimensional structures reported place the enzyme in the large Gcn5-related N-acetyltransferase (GNAT) superfamily. The structure of the CoA-ribostamycin ternary complex allows us to propose a chemical mechanism for the reaction, and comparison with the Mycobacterium tuberculosis AAC(2')-CoA ribostamycin complex allows us to define how regioselectivity of acetylation is achieved. The AAC(6')-Iy dimer is most structurally similar to the Saccharomyces cerevisiae Hpa2-encoded histone acetyltransferase. We demonstrate that AAC(6')-Iy catalyzes both acetyl-CoA-dependent self-alpha-N-acetylation and acetylation of eukaryotic histone proteins and the human histone H3 N-terminal peptide. These structural and catalytic similarities lead us to propose that chromosomally encoded bacterial acetyltransferases, including those functionally identified as aminoglycoside acetyltransferases, are the evolutionary progenitors of the eukaryotic histone acetyltransferases. PMID- 15123252 TI - Transformation of aminoacyl tRNAs for the in vitro selection of "drug-like" molecules. AB - Evolutionary approaches are regularly used to isolate single molecules with desired activities from large populations of nucleic acids (approximately 10(15)). Several methods have also been developed to generate libraries of mRNA encoded peptides and proteins for the in vitro selection of functional polypeptides. In principal, such mRNA encoding systems could be used with libraries of nonbiological polymers if the ribosome can be directed to polymerize tRNAs carrying unnatural amino acids. The fundamental problem is that current chemical aminoacylation systems cannot easily produce sufficient amounts of the numerous misacylated tRNAs required to synthesize a complex library of encoded polymers. Here, we show that bulk-aminoacylated tRNA can be transformed into N monomethylated aminoacyl tRNA and translated. Because poly-N-methyl peptide backbones are refractory to proteases and are membrane permeable, our method provides an uncomplicated means of evolving novel drug candidates. PMID- 15123253 TI - Benitec, Ltd. Gene silencing from down under. PMID- 15123254 TI - Optimized chimeragenesis; creating diverse p450 functions. AB - A drawback to generating chimeric proteins by chimeragenesis, especially when the "parent" proteins share low sequence identity, is that unfolded proteins frequently result. In this issue of Chemistry & Biology, Arnold and coworkers report their use of the SCHEMA algorithm to effectively predict ideal hybrids of cytochromes p450. PMID- 15123255 TI - Quantifying intermediates in template-directed natural product biosynthesis. AB - High-performance mass spectrometry is providing new experimental windows into the enzymology of natural product biosynthesis. The first quantitative assessments of covalently attached biosynthetic intermediates promise to shine new light on template-directed biosynthesis. PMID- 15123256 TI - Profiling natural product biosynthesis. AB - Natural products are a rich source of therapeutics; however, artificially reengineering the biosynthetic pathways that generate these compounds could potentially generate "designer" drugs. Last month in Chemistry & Biology, Burkart and coworkers reported their technique to track and better understand the components of these pathways. PMID- 15123257 TI - The modular approach to ligand discovery. AB - Identifying specific protein-ligand interactions is a long-standing problem in drug discovery and chemical biology, which is only exacerbated by the abundance of uncharacterized proteins revealed by genomics. Last month in Chemistry Biology, Sem et al. described a powerful technique for rapidly screening protein families for ligands. PMID- 15123258 TI - Rpd3p relocation mediates a transcriptional response to rapamycin in yeast. AB - Treating yeast cells with rapamycin, a small molecule that inhibits the TOR proteins, leads to the repression of many genes. Consistent with prior studies, we find that RPD3, which encodes a histone deacetylase (HDAC), is required for repression upon rapamycin treatment. To elucidate the mechanism underlying RPD3 mediated repression, we screened all promoters in yeast for occupancy by Rpd3p before and after treatment with rapamycin. We find that Rpd3p binds to the promoters of rapamycin-repressible genes only following treatment. These data conflict with a previously proposed model suggesting that Rpd3p is constitutively bound to rapamycin-repressible genes and becomes active only after a stimulus such as treatment with rapamycin. Rather, the comprehensive analysis presented here strongly supports a model in which recruitment of Rpd3p to gene promoters is a regulated step in the control of gene repression. PMID- 15123259 TI - Cellular delivery of MRI contrast agents. AB - Magnetic resonance imaging (MRI) is a powerful tool for acquiring images of opaque living animals with the benefit of tracking events over extended periods of time on the same specimen. Contrast agents are used to enhance regions, tissues, and cells that are magnetically similar but histologically distinct. A principal barrier to the development of MRI contrast agents for investigating biological questions is the delivery of agents across cellular membranes. Here, we describe the synthesis and in vitro testing of Gd(III)-based MRI contrast agents containing varying length polyarginine oligomers capable of permeating cell membranes. We examine the effect of the length of oligomer on T(1) enhancement and cellular uptake. Furthermore, the effect of incubation time, concentration, and cell type on uptake is explored. Toxicity and washout studies are performed in addition to MRI phantom studies. PMID- 15123260 TI - Functional evolution and structural conservation in chimeric cytochromes p450: calibrating a structure-guided approach. AB - Recombination generates chimeric proteins whose ability to fold depends on minimizing structural perturbations that result when portions of the sequence are inherited from different parents. These chimeric sequences can display functional properties characteristic of the parents or acquire entirely new functions. Seventeen chimeras were generated from two CYP102 members of the functionally diverse cytochrome p450 family. Chimeras predicted to have limited structural disruption, as defined by the SCHEMA algorithm, displayed CO binding spectra characteristic of folded p450s. Even this small population exhibited significant functional diversity: chimeras displayed altered substrate specificities, a wide range in thermostabilities, up to a 40-fold increase in peroxidase activity, and ability to hydroxylate a substrate toward which neither parent heme domain shows detectable activity. These results suggest that SCHEMA-guided recombination can be used to generate diverse p450s for exploring function evolution within the p450 structural framework. PMID- 15123261 TI - Crosslinking of and coupling to viral capsid proteins by tyrosine oxidation. AB - Cowpea mosaic virus is composed of 60 identical copies of a two-subunit protein organized in pentameric assemblies around the icosahedral 5-fold symmetry axis. Treatment of the virus with the Ni(II) complex of the tripeptide GGH and a peroxide oxidant, or irradiation in the presence of Ru(bpy)(3)(2+) and persulfate generates covalent crosslinks across the pentameric subunit boundaries, effectively stitching the subunits together. Intersubunit crosslinking was found to occur exclusively at adjacent tyrosine residues (Y52-Y103), as predicted from the X-ray crystal structure of the capsid, and to be more extensive with the photochemical ruthenium system. The Ni/GGH oxidative procedure was also used to make covalent attachments to the virion by trapping with a functionalized disulfide reagent. PMID- 15123262 TI - Mass spectrometric interrogation of thioester-bound intermediates in the initial stages of epothilone biosynthesis. AB - Direct detection of thioester intermediate mixtures bound to EpoC, a 195 kDa polyketide synthase, has been achieved using limited proteolysis and Fourier transform mass spectrometry (FTMS). Incubation with various N-acetylcysteamine thioester (S-NAC) substrate mimics produced mass shifts on the EpoC ACP domain consistent with their condensation with an enzyme-bound carbanion produced by the decarboxylation of methylmalonyl-S-EpoC. Reconstitution of EpoA-ACP, EpoB, and EpoC gave a +165.0 Da mass shift consistent with the formation of the methylthiazolyl-methacrylyl product by incorporation of acetyl-CoA, cysteine, and methylmalonyl-CoA. Thioester-templated reaction intermediates and products are typically characterized by quantifying radioactive substrates, either enzyme bound or chemically hydrolyzed. In contrast, the MS-based methodology described here provides semiquantifiable ratios of free enzyme, intermediate, and product occupancy and reveals that certain substrates result in a >50% formation of nonproductive intermediates. PMID- 15123263 TI - Small molecule inhibitors of mucin-type O-linked glycosylation from a uridine based library. AB - The polypeptide N-acetyl-alpha-galactosaminyltransferases (ppGalNAcTs, also abbreviated ppGaNTases) initiate mucin-type O-linked glycosylation and therefore play pivotal roles in cell-cell communication and protection of tissues. In order to develop new tools for studying mucin-type O-linked glycosylation, we screened a 1338 member uridine-based library to identify small molecule inhibitors of ppGalNAcTs. Using a high-throughput enzyme-linked lectin assay (ELLA), two inhibitors of murine ppGalNAcT-1 (K(I) approximately 8 microM) were identified that also inhibit several other members of the family. The compounds did not inhibit other mammalian glycosyltransferases or nucleotide sugar utilizing enzymes, suggesting selectivity for the ppGalNAcTs. Treatment of cells with the compounds abrogated mucin-type O-linked glycosylation but not N-linked glycosylation and also induced apoptosis. These uridine analogs represent the first generation of chemical tools to study the functions of mucin-type O-linked glycosylation. PMID- 15123264 TI - In vivo targeting of organic calcium sensors via genetically selected peptides. AB - A library of constrained peptides that form stable folded structures was screened for aptamers that bind with high affinity to the fluorescent dye Texas red. Two selected clones had binding constants to Texas red of 25 and 80 pM as phage and binding had minimal effects on the fluorescence of Texas red. The peptides interact with distinct but overlapping regions of Texas red. One peptide bound to X-rhod calcium sensors, which share the same core fluorophore as Texas red. These dyes retained calcium sensitivity when bound to the peptide. This peptide was used to label a fusion protein with X-rhod-5F in vivo, and X-rhod sensed changes in calcium locally. Thus, minimal, constrained peptides can functionally bind to environmentally sensitive dyes or other organic agents in biological contexts, suggesting tools for in vivo imaging and analysis. PMID- 15123265 TI - Starter unit choice determines the production of two tetraene macrolides, rimocidin and CE-108, in Streptomyces diastaticus var. 108. AB - Streptomyces diastaticus var. 108, a newly isolated strain, produces two closely related tetraene macrolides (rimocidin and CE-108) as well as oxytetracycline. A region of 19,065 base pairs of DNA from the S. diastaticus var. 108 genome was isolated, sequenced, and characterized. Ten complete genes and one truncated ORF were located. Disruption of these genes proved that this genomic region is part of the biosynthetic cluster for the two tetraenes. The choice of starter units by the loading module and the in vivo availability of the starter metabolites are crucial for the final ratio of the two macrolides. A second type I PKS, unrelated to tetraene biosynthesis, was also identified; disruption of these genes suggests that they would code for enzymes involved in the biosynthesis of a polyketide that might compete metabolically with rimocidin production. PMID- 15123266 TI - Novel heparan sulfate mimetic compounds as antitumor agents. AB - Heparan sulfate glycosaminoglycans (HSGAGs) are involved in tumor cell growth, adhesion, invasion, and migration, due to their interactions with various proteins. In this study, novel HSGAG-mimetic compounds (KI compounds) were designed and synthesized. As a result of cell-based assays, KI-105 was found to exert potent inhibitory activities against migration and invasion of human fibrosarcoma HT1080 cells. The present results indicate that a novel invasion/migration inhibitor, KI-105, can increase the adherence of HT1080 cells. It was conceivable that this cellular effect was caused by an increase in the amount of cell-surface HSGAGs and focal adhesions. Although further investigations are needed to decipher the molecular mechanism of KI-105, it is suggested that heparanase and Cdc42 are involved in its biological effects. PMID- 15123267 TI - Cytochrome p450 taxadiene 5alpha-hydroxylase, a mechanistically unusual monooxygenase catalyzing the first oxygenation step of taxol biosynthesis. AB - The first oxygenation step in the biosynthesis of the anticancer drug taxol in Taxus species is the cytochrome p450-mediated hydroxylation (with double bond migration) of the diterpene olefin precursor taxa-4(5),11(12)-diene to taxa 4(20),11(12)-dien-5alpha-ol. A homology-based cloning strategy, employing an induced Taxus cell library, yielded a cDNA encoding taxadiene 5alpha-hydroxylase, which was functionally expressed in yeast and insect cells. The recombinant enzyme was characterized and shown to efficiently utilize both taxa-4(5),11(12) diene and taxa-4(20),11(12)-diene (as an adventitious substrate) to synthesize taxa-4(20),11(12)-dien-5alpha-ol. This hydroxylase resembles, in sequence and properties, other cytochrome p450 oxygenases of taxol biosynthesis. The utilization of both taxadiene isomers in the formation of taxa-4(20),11(12)-dien 5alpha-ol is novel, suggesting a reaction mechanism involving promiscuous radical abstraction with selective oxygen insertion rather than epoxidation of the C4,C5 alkene of the natural substrate and allylic rearrangement of the resulting taxa 11(12)-en-4,5epoxide. PMID- 15123268 TI - Rational design and real time, in-cell detection of the proapoptotic activity of a novel compound targeting Bcl-X(L). AB - Antiapoptotic Bcl-2-family proteins Bcl-2 and Bcl-X(L) have been recently validated as drug discovery targets for cancer. Here, by using a combination of molecular modeling, NMR-based structural analysis, fluorescence polarization assays, and cell-based assays, we have designed and characterized a novel proapoptotic compound targeting these proteins. Our compound, Apogossypol, is capable of binding and inhibiting Bcl-2 and Bcl-X(L) with high affinity and induces apoptosis of tumor cell lines. Mechanistic studies on the action of our compound were also performed via confocal microscopy that provided real-time detection of the interaction with Bcl-X(L) in intact cells. Finally, preliminary data on cells freshly isolated from patients affected by chronic lymphocytic leukemia strongly suggest potential applications of Bcl-2 antagonists as chemosensitizers in cancer therapy. PMID- 15123269 TI - A new class of phytoestrogens; evaluation of the estrogenic activity of deoxybenzoins. AB - Although deoxybenzoins are intermediates in the synthesis of isoflavones, their estrogenic activity has not been investigated. Eleven deoxybenzoins were synthesized and their estrogenicity was evaluated. While their affinities for estrogen receptors (ER) ERalpha and ERbeta were found grossly comparable to those of daidzein, some exhibited considerable selectivity and transcriptional bias toward ERbeta, which appeared to allow for enhancement of ER-mediated transcription via deoxybenzoin binding of ERbeta. Their activity to stimulate the proliferation of ER-positive breast cancer cells and regulate the expression of endogenous and stably transfected reporter genes differed considerably, with some inhibiting cell proliferation while effectively inducing gene expression at the same time. Molecular modeling confirmed that deoxybenzoins fit well in the ligand binding pocket of ERbeta, albeit with different orientations. Our data support the view that deoxybenzoins constitute a promising new class of ERbeta-biased phytoestrogens. PMID- 15123270 TI - Identification of small molecule inhibitors that distinguish between non transferrin bound iron uptake and transferrin-mediated iron transport. AB - Chemical genetics is an emerging field that takes advantage of combinatorial chemical and small molecule libraries to dissect complex biological processes. Here we establish a fluorescence-based assay to screen for inhibitors of iron uptake by mammalian cells. Using this approach, we screened the National Cancer Institute's Diversity Set library for inhibitors of non-transferrin bound iron uptake. This screen identified 10 novel small molecule inhibitors of iron transport with IC(50) values that ranged from 5 to 30 microM. Of these ten compounds, only two blocked uptake of iron mediated by transferrin. Thus, this study characterizes the first small molecule inhibitors that distinguish between different pathways of iron transport. PMID- 15123271 TI - Thios pharmaceuticals. Targeting sulfation pathways. PMID- 15123272 TI - RNA: primed for packing? AB - How does RNA's seemingly subtle 2'-hydroxyl group contribute to its diverse biological functions? Using quantitative structure-activity relationship (QSAR) analysis, Piccirilli and colleagues find that the 2'-OH fosters van der Waals and hydrophobic interactions that play underappreciated roles in RNA structure and molecular recognition. PMID- 15123273 TI - Finding Cinderella after the ball: a three-hybrid approach to drug target identification. AB - A major bottleneck in drug discovery is identifying the targets of small molecules. The yeast three-hybrid assay extends the two-hybrid approach to screen for protein-small molecule interactions. In this issue of Chemistry & Biology, GPC Biotech reports the first application of this promising assay. PMID- 15123274 TI - What makes epothilones stick? AB - The potent epothilone tubulin polymerization promoters have been studied extensively by synthetic and SAR approaches. The paper by Buey et al. (, this issue of Chemistry & Biology) adds a new depth of mechanistic understanding by a careful analysis of the tubulin polymerization mechanism of the epothilones. PMID- 15123275 TI - New start and finish for complex polyketide biosynthesis. AB - The polyketide vicenistatin has significant anticancer activity. In the January issue of Chemistry & Biology, Kakinuma and coworkers reported on the vicenistatin biosynthetic gene cluster and demonstrated in vitro glycosylation of its aglycone core, raising the possibility of producing analogs with altered sugar residues. PMID- 15123276 TI - Understanding diseases via receptor regulation. AB - In this issue of Chemistry & Biology, Guy and coworkers demonstrate that they can selectively recruit individual nuclear receptors by using small molecules (proteomimetics) in combination with specific agonists. This may ultimately lead to a link between the receptor's signaling pathway and its role in individual diseases. PMID- 15123277 TI - How to silence silencing. AB - Two recent reports describe the stunning crystal structures of complexes between a viral protein that suppresses RNA silencing and a 21 nucleotide small interfering (si)RNA. PMID- 15123278 TI - Toxicogenomics in predictive toxicology in drug development. AB - The goal of toxicology is the assessment of possible risk to man. An emerging technology with the potential to have a major impact on risk assessment is toxicogenomics. In this review, we provide an overview of the many possibilities for toxicogenomics including technology platforms, data interpretation, and regulatory perspective and we give examples of toxicogenomics investigations. Toxicogenomics is a powerful tool for compound classification, for mechanistic studies, and for the detection of toxicity markers. Thus, toxicogenomics helps in the extrapolation of findings across species and increases predictability. Biomarkers are valuable in the evaluation of compounds at earlier development phases, improving clinical candidate selection. Caution regarding the interpretation of the results is still necessary. Nevertheless, toxicogenomics will accelerate preclinical safety assessments and improve the prediction of toxic liabilities, as well as of potential risk accumulation for drug-drug or drug-disease interactions. PMID- 15123279 TI - In vitro and in vivo production of new aminocoumarins by a combined biochemical, genetic, and synthetic approach. AB - The aminocoumarin antibiotics clorobiocin, novobiocin, and coumermycin A(1) are inhibitors of bacterial gyrase. Their chemical structures contain amide bonds, formed between an aminocoumarin ring and an aromatic acyl component, which is 3 dimethylallyl-4-hydroxybenzoate in the case of novobiocin and clorobiocin. These amide bonds are formed under catalysis of the gene products of cloL, novL, and couL, respectively. We first examined the substrate specificity of the purified amide synthetases CloL, NovL, and CouL for the various analogs of the prenylated benzoate moiety. We then generated new aminocoumarin antibiotics by feeding synthetic analogs of the 3-dimethylallyl-4-hydroxybenzoate moiety to a mutant strain defective in the biosynthesis of the prenylated benzoate moiety. This resulted in the formation of 32 new aminocoumarin compounds. The structures of these compounds were elucidated using FAB-MS and (1)H-NMR spectroscopy. PMID- 15123280 TI - Systems-based design of bi-ligand inhibitors of oxidoreductases: filling the chemical proteomic toolbox. AB - Genomics-driven growth in the number of enzymes of unknown function has created a need for better strategies to characterize them. Since enzyme inhibitors have traditionally served this purpose, we present here an efficient systems-based inhibitor design strategy, enabled by bioinformatic and NMR structural developments. First, we parse the oxidoreductase gene family into structural subfamilies termed pharmacofamilies, which share pharmacophore features in their cofactor binding sites. Then we identify a ligand for this site and use NMR-based binding site mapping (NMR SOLVE) to determine where to extend a combinatorial library, such that diversity elements are directed into the adjacent substrate site. The cofactor mimic is reused in the library in a manner that parallels the reuse of cofactor domains in the oxidoreductase gene family. A library designed in this manner yielded specific inhibitors for multiple oxidoreductases. PMID- 15123281 TI - Manipulation of carrier proteins in antibiotic biosynthesis. AB - Engineering biosynthetic pathways into suitable host organisms has become an attractive venue for the design, evaluation, and production of small molecule therapeutics. Polyketide (PK) and nonribosomal peptide (NRP) synthases have been of particular interest due to their modular structure, yet routine cloning and expression of these enzymes remains challenging. Here we describe a method to covalently label carrier proteins from PK and NRP synthases using the enzymatic transfer of a modified coenzyme A analog by a 4'-phosphopantetheinyltransferase. Using this method, carrier proteins can be loaded with single fluorescent or affinity reporters, providing novel entry for protein visualization, Western blot identification, and affinity purification. Application of these methods provides an ideal tool to track and quantify metabolically engineered pathways. Such techniques are valuable to measure protein expression, solubility, activity, and native posttranslational modification events in heterologous systems. PMID- 15123282 TI - Bright fluorescent chemosensor platforms for imaging endogenous pools of neuronal zinc. AB - A series of new fluorescent Zinpyr (ZP) chemosensors based on the fluorescein platform have been prepared and evaluated for imaging neuronal Zn(2+). A systematic synthetic survey of electronegative substitution patterns on a homologous ZP scaffold provides a basis for tuning the fluorescence responses of "off-on" photoinduced electron transfer (PET) probes by controlling fluorophore pK(a) values and attendant proton-induced interfering fluorescence of the metal free (apo) probes at physiological pH. We further establish the value of these improved optical tools for interrogating the metalloneurochemistry of Zn(2+); the novel ZP3 fluorophore images endogenous stores of Zn(2+) in live hippocampal neurons and slices, including the first fluorescence detection of Zn(2+) in isolated dentate gyrus cultures. Our findings reveal that careful control of fluorophore pK(a) can minimize proton-induced fluorescence of the apo probes and that electronegative substitution offers a general strategy for tuning PET chemosensors for cellular studies. In addition to providing improved optical tools for Zn(2+) in the neurosciences, these results afford a rational starting point for creating superior fluorescent probes for biological applications. PMID- 15123283 TI - A three-hybrid approach to scanning the proteome for targets of small molecule kinase inhibitors. AB - In this study, we explored the application of a yeast three-hybrid (Y3H)-based compound/protein display system to scanning the proteome for targets of kinase inhibitors. Various known cyclin-dependent kinase (CDK) inhibitors, including purine and indenopyrazole analogs, were displayed in the form of methotrexate based hybrid ligands and deployed in cDNA library or yeast cell array-based screening formats. For all inhibitors, known cell cycle CDKs as well as novel candidate CDK-like and/or CDK-unrelated kinase targets could be identified, many of which were independently confirmed using secondary enzyme assays and affinity chromatography. The Y3H system described here may prove generally useful in the discovery of candidate drug targets. PMID- 15123284 TI - Interaction of epothilone analogs with the paclitaxel binding site: relationship between binding affinity, microtubule stabilization, and cytotoxicity. AB - The interactions of epothilone analogs with the paclitaxel binding site of microtubules were studied. The influence of chemical modifications in the C15 side chain and in C12 on binding affinity and microtubule elongation was characterized. Modifications favorable for binding affinity are (1). a thiomethyl group at C21 of the thiazole side chain, (2). a methyl group at C12 in S configuration, (3). a pyridine side chain with C15 in S configuration, and (4). a cyclopropyl moiety between C12 and C13. The same modification in different ligands has similar effect on affinity, allowing good structure-affinity characterization. The correlation between binding, microtubule stabilization, and cytotoxicity of the compounds has been determined, showing differential effects of the modifications. The binding constants correlate well with IC(50) values, demonstrating that affinity measurements are a useful tool for drug design. PMID- 15123285 TI - New strategies for exploring RNA's 2'-OH expose the importance of solvent during group II intron catalysis. AB - The 2'-hydroxyl group contributes inextricably to the functional behavior of many RNA molecules, fulfilling numerous essential chemical roles. To assess how hydroxyl groups impart functional behavior to RNA, we developed a series of experimental strategies using an array of nucleoside analogs. These strategies provide the means to investigate whether a hydroxyl group influences function directly (via hydrogen bonding or metal ion coordination), indirectly (via space filling capacity, inductive effects, and sugar conformation), or through interactions with solvent. The nucleoside analogs span a broad range of chemical diversity, such that quantitative structure activity relationships (QSAR) now become possible in the exploration of RNA biology. We employed these strategies to investigate the spliced exons reopening (SER) reaction of the group II intron. Our results suggest that the cleavage site 2'-hydroxyl may mediate an interaction with a water molecule. PMID- 15123286 TI - Synthesis and target identification of hymenialdisine analogs. AB - Hymenialdisine (HMD) is a sponge-derived natural product kinase inhibitor with nanomolar activity against CDKs, Mek1, GSK3beta, and CK1 and micromolar activity against Chk1. In order to explore the broader application of the pyrrolo[2,3 c]azepine skeleton of HMD as a general kinase inhibitory scaffold, we searched for additional protein targets using affinity chromatography in conjunction with the synthesis of diverse HMD analogs and profiled HMD against a panel of 60 recombinant enzymes. This effort has led to nanomolar to micromolar inhibitors of 11 new targets including p90RSK, KDR, c-Kit, Fes, MAPK1, PAK2, PDK1, PKCtheta, PKD2, Rsk1, and SGK. The synthesis of HMD analogs has resulted in the identification of compounds with enhanced and/or dramatically altered selectivities relative to HMD (28n) and in molecules with antiproliferative activities 30-fold higher than HMD (28p). PMID- 15123287 TI - Rational design of a bimodular model system for the investigation of heterocyclization in nonribosomal peptide biosynthesis. AB - Cyclization (Cy) domains in NRPS catalyze the heterocyclization of cysteine and serine/threonine to thiazoline and oxazoline rings. A model system consisting of the first two modules of bacitracin synthetase A fused to the thioesterase (Te) domain of tyrocidine synthetase was constructed (BacA1-2-Te) and shown to be active in production of the heterocyclic IleCys(thiazoline). Based on this model system, the feasibility of Cy domain module fusions was investigated by replacing the BacA2 Cy-A-PCP-module with modules of MbtB and MtaD from the biosynthesis systems of mycobactin and myxothiazol, revealing the formation of novel heterocyclic dipeptides. To dissect the reaction sequence of the Cy domain in peptide bond formation and heterocyclization, several residues of the BacA1-2-Te Cy domain were analyzed by mutagenesis. Two mutants exhibited formation of the noncyclic dipeptide, providing clear evidence for the independence of condensation and cyclization. PMID- 15123288 TI - Ligand-selective inhibition of the interaction of steroid receptor coactivators and estrogen receptor isoforms. AB - Ligand-dependent nuclear hormone receptor (NR) signaling requires direct interaction between NR and the steroid receptor coactivators (SRC). Herein we utilize a library of SRC2 peptidomimetics to select for specific inhibitors of the interaction of SRC2 with the two estrogen receptor (ER) isoforms, ERalpha and ERbeta, in the presence of three different ligands: 17beta-estradiol, diethylstilbesterol, and genistein. The pattern of inhibitor selectivity for each ER isoform varied depending upon which ligand was present, thus demonstrating that the ligands exert unique allosteric effects upon the surface of the SRC binding pocket. Several of the lead compounds are highly (>100-fold) selective for blocking the binding of SRC2 to ERalpha, in preference to ERbeta, in the presence of one ligand and therefore may prove useful for decoupling ERbeta signaling from ERalpha signaling. PMID- 15123289 TI - Efficacy of cellular vaccines and genetic adjuvants against bacterial kidney disease in chinook salmon (Oncorhynchus tshawytscha). AB - DNA adjuvants and whole bacterial cell vaccines against bacterial kidney disease (BKD) were tested in juvenile chinook salmon. Whole cell vaccines of either a nonpathogenic Arthrobacter spp. or an attenuated Renibacterium salmoninarum strain provided limited prophylactic protection against acute intraperitoneal challenge with virulent R. salmoninarum, and the addition of either synthetic oligodeoxynucleotides or purified R. salmoninarum genomic DNA as adjuvants did not increase protection. However, a combination of both whole cell vaccines significantly increased survival among fish naturally infected with R. salmoninarum, and the surviving fish treated with the combination vaccine exhibited reduced levels of bacterial antigens in the kidney. This is the first demonstration of a potential therapeutic effect of a whole cell vaccine against BKD. PMID- 15123290 TI - Vitamin E requirements of juvenile grass shrimp, Penaeus monodon, and effects on non-specific immune responses. AB - A feeding trial was conducted to determine the dietary vitamin E (DL-alpha tocopheryl acetate, dl-alpha-TOA) requirement and its effect on the non-specific immune responses of juvenile grass shrimp, Penaeus monodon. Purified diets with eight levels (0, 25, 50, 75, 100, 150, 200, 400 mg vitamin E kg diet-1) of supplemental dl-alpha-TOA were fed to P. monodon (mean initial weight 0.29 +/- 0.01 g) for eight weeks. Each diet was fed to three replicate groups of shrimp. Weight gains and total haemocyte count (THC) were higher (P < 0.05) in shrimp fed diets supplemented with 75 and 100 mg vitamin E kg diet-1 than in shrimp fed diets supplemented with 60-fold induction compared to supernatants from non-stimulated cells). There was no response to supernatants from LPS- and ConA/PMA-stimulated leucocytes, demonstrating the specificity for type I interferon-like activity. Duplicate samples analysed using a cell protection assay for detection of antiviral activity correlated well with levels obtained by the Mx1 promoter reporter gene assay (R2=0.97), confirming the reporter assay as a reliable substitute for the standard antiviral assay. The Mx reporter gene assay also has advantages in terms of sensitivity, high dynamic range and reliability over the conventional cell protection assay. PMID- 15123322 TI - Influences of traditional Chinese medicine on non-specific immunity of Jian Carp (Cyprinus carpio var. Jian). AB - The influence of traditional Chinese medicine (TCM) formulation from Astragalus Root (Radix astragalin seu Hedysari) and Chinese Angelica Root (R. Angelicae Sinensis) at a ratio of 5:1 (w/w) on non-specific immunity of Jian carp, Cyprinus carpio var. Jian was investigated. The number of NBT-positive cells in the blood and lysozyme and complement activities in the serum of Carp fed with commercial feed supplemented with 1.0% (diet 1) and 1.5% (diet 2) TCM at 10 day of post feeding were not different from those of the control group fed with feed unsupplemented TCM 10 days post-feeding (P>0.05), but at 20 and 30 days they increased significantly (P<0.05). The values of diet 1 group and diet 2 group at 20 day and at 30 day were not significantly different (P>0.05) from each other. In addition, the TCM formula increased body weight of experimental fish by about 16.84% (diet 1) and 19% (diet 2) above that of the control group. Therefore, these data suggest that the TCM formula could elevate the function of non specific immunity of Jian carp. The optimal dosage added to commercial carp feed was 1.0% (w/w) and the oral administration time as a course of treatment was 20 days. PMID- 15123323 TI - The auxotrophic aroA mutant of Aeromonas hydrophila as a live attenuated vaccine against A. salmonicida infections in rainbow trout (Oncorhynchus mykiss). AB - An auxotrophic aroA mutant of the Aeromonas hydrophila AG2 strain is a live attenuated vaccine against A. hydrophila infection in rainbow trout (Oncorhynchus mykiss). The protection conferred by the live attenuated vaccine against A. salmonicida strains is reported here, and several parameters of the specific and non-specific immune response in vaccinated trout were characterised. Vaccination with a dose of 10(7)cells/fish of the aroA mutant elicited significant protection against the Hooke and DK30 strains of A. salmonicida (relative percent survival RPS >60%). This cross-protection correlated moderately with the activation of the humoral and cellular specific immune responses, which show cross-reactivity against antigens shared by the two bacterial species, and a moderate increase in the lysozyme and antiprotease activities in the serum of vaccinated trout. PMID- 15123324 TI - Role of capsule in serotypic differences and complement fixation by Lactococcus garvieae. AB - Seventeen geographically distinct isolates of Lactococcus garvieae, isolated from diseased fish, were compared serologically using antiserum raised against the various isolates in rainbow trout. Sera raised against a capsule deficient isolate did not agglutinate capsulated isolates, regardless of origin. In contrast, all antisera raised against capsulated isolates cross reacted strongly with non-capsulated isolates. Antisera raised against capsulated Japanese isolates cross reacted with other capsulated Japanese isolates including isolates from geographically distinct prefectures within Japan (Ehime and Oita). However, antisera against these virulent capsulated isolates did not cross react with European capsulated isolates. Antisera raised against European capsulated isolates cross reacted with other European isolates, regardless of origin within Europe (UK, Italy, Spain), but did not cross-react with Japanese capsulated isolates. Agglutination assays performed with a range of fifteen lectins revealed differences in surface carbohydrate structure: capsule deficient isolates agglutinated with concanavalin A, Ricinis communis agglutinin, Pisum sativum agglutinin, Lens culinaris agglutinin, wheat germ agglutinin and succinylated wheat germ agglutinin. European capsulated isolates agglutinated with concanavalin A only. The Japanese capsulated isolates were not agglutinated by any of the lectins used in this study. Representative isolates from each group (Japanese capsulated and non-capsulated, European capsulated and non-capsulated) were investigated for their ability to fix complement. Non-capsulated isolates fixed complement regardless of origin, and antibody did not markedly enhance complement fixation. In contrast, the capsulated isolates were less efficient at fixing complement, but complement fixation was markedly increased by homologous antibody. PMID- 15123325 TI - In vitro effects of LPS, IL-2, PDGF and CRF on haemocytes of Mytilus galloprovincialis Lmk. AB - The cells in charge of the innate immune response in the marine mussel Mytilus galloprovincialis Lmk. are the haemocytes. These cells respond in different ways to agents such as lipopolysaccharide (LPS), interleukin-2 (IL-2), platelet derived growth factor (PDGF) and corticotropin releasing factor (CRF). After stimulation of the haemocytes, the expression of molecules reactive with monoclonal antibodies raised to the alpha chain of the IL-2 receptor, present in their membrane, differed depending on the agent used. The same happened with regard to the levels of dopamine, adrenaline and noradrenaline released to the medium by the haemocytes. It should also be noted that no catecholamine release was detected and the level of expression of IL-2Ralpha showed no significant variation in cultured cells that had not been treated with inducers. These facts would indicate that most haemocytes were in the same starting condition at the moment that the stimulation was performed. Therefore, cultured haemocytes can be a highly reliable model in the study of the innate immune system. PMID- 15123326 TI - Isolation and characterization of complement component C3 from Atlantic cod (Gadus morhua L.) and Atlantic halibut (Hippoglossus hippoglossus L.). AB - Complement component C3 was isolated from the plasma of cod (Gadus morhua L.) and halibut (Hippoglossus hippoglossus L.). Fast protein liquid chromatography (FPLC) techniques, involving ion exchange and gel filtration columns, were used. The purified proteins were analysed by SDS-PAGE which showed a two-chain structure, alpha- and beta-chains, as seen in higher vertebrates. Both proteins had intra chain thioesters located within their alpha-chains and N-terminal amino acid sequencing confirmed their identity with reference to known C3 amino acid sequences from other species. Specific antibodies were prepared against cod and halibut C3 and tested in Western blotting on sera and purified C3. The proteolytic fragmentation of C3 was tested with trypsin, pepsin, papain and the extracellular product (ECP) from the bacterium Aeromonas salmonicida ssp. achromogenes (Asa). Both trypsin and papain were successful in cleaving C3 whereas pepsin and ECP had no effect. Carbohydrate moieties were detected in the alpha- and beta-chains of cod and halibut C3 and N-linked oligosaccharides were removed from the C3 with PNGase treatment, revealing a difference in C3 glycosylation between the two species. PMID- 15123327 TI - The effect of dietary administration of the fungus Mucor circinelloides on non specific immune responses of gilthead seabream. AB - The immunostimulant potential of the whole fungus Mucor circinelloides administered in the diet to gilthead seabream (Sparus aurata L.) was studied. Three different lyophilised strains were used: the wild-type (R7B) and two mutant strains in the carotene-production pathway which are enriched in lycopene (MU224) or beta-carotene (T31). Fish were fed diets containing 0 (control) or 10 g of one of the whole M. circinelloides strains per kg feed. After 2, 4 or 6 weeks of treatment the growth rate as well as humoral (lysozyme activity) and cellular (phagocytosis and cytotoxicity) immune responses were determined. The specific growth rate increased slightly with all the M. circinelloides-supplemented diets. Serum lysozyme activity increased slightly (P>0.05) in fish fed the mutant strain supplemented diets. Of the cellular responses, phagocytosis significantly increased after 6 weeks, in fish fed the wild-type strain-supplemented diets while cytotoxicity increased after 4 weeks in fish fed the beta-carotene-enriched strain-supplemented diet. The immune responses were increased to some degree by diets containing whole M. circinelloides strains. These results are discussed in the light of the on-going search for new microorganisms, wild or mutant-type, for use as immunostimulants in fish farming. PMID- 15123328 TI - Expression of luciferase in selected organs following delivery of naked and formulated DNA to rainbow trout (Oncorhynchus mykiss) by different routes of administration. AB - In the present work, the expression of luciferase in selected organs following administration of DNA delivered as naked, liposome-formulated or chitosan formulated by different routes of administration (intramuscular, intraperitoneal and intravenous injection, immersion and anal intubation) was studied in rainbow trout (Oncorhynchus mykiss). The different formulations and routes of administration both influenced in which organs luciferase was expressed and the magnitude of expression. The highest expression levels of luciferase in the head kidney and liver were found after an intraperitoneal injection of lipoplex 2. In the spleen, the highest levels were detected after injection of naked DNA (intraperitonal or intramuscular) and lipoplex 2 (intraperitoneal). Following intravenous injection, naked DNA gave higher expression levels in the organs than the formulated plasmids and immersion and anal intubation were not effective routes of delivery as no expression of luciferase could be detected in any of the organs tested. Additionally, PCR using a primer specific for a 600 bp region of the luciferase gene pcDNA3-luc was used to assess the distribution of the plasmid itself after intramuscular and intraperitoneal injection. Positive amplification was obtained in spleen, head kidney, liver and muscle at the injection site following injection of formulated plasmids, while only muscle tissue from the injection site was positive when naked DNA was used. PMID- 15123329 TI - Is Apolipoprotein A-I a regulating protein for the complement system of cod (Gadus morhua L.)? PMID- 15123331 TI - Neurochemical diagnosis of Alzheimer's dementia by CSF Abeta42, Abeta42/Abeta40 ratio and total tau. AB - Cerebrospinal fluid (CSF) concentrations of amyloid beta peptides ending at positions 42 and 40 (Abeta42 and Abeta40, respectively), and total tau (tTau) protein were measured by ELISA in order to compare their accuracy in discriminating patients with Alzheimer's disease (AD, n = 22), non-Alzheimer dementia (nAD, n = 11) and control subjects (CON, n=35). As compared to the other groups, the concentrations of Abeta42 and tTau were decreased (P<0.001) and increased (P<0.001) in AD, respectively, while Abeta40 did not differ significantly among the groups. Receiver operating characteristic (ROC) analysis was performed to define cut-off values for maximized sensitivity and specificity. For all groups compared the Abeta peptide ratio 42/40 classified more patients correctly, as compared to the concentration of Abeta42 alone: AD versus controls, 94 and 86.7%; AD versus nAD, 90 and 85% and AD versus nAD plus controls, 90.8 and 87%, respectively. The percentage of correctly classified patients was further improved when the Abeta ratio was combined with the analysis of the tTau concentration. Presence of the apolipoprotein E 4 allele, age or degree of mental disability did not significantly influence the parameters studied. PMID- 15123332 TI - Co-accumulation of vascular endothelial growth factor with beta-amyloid in the brain of patients with Alzheimer's disease. AB - Alzheimer's disease (AD) is accompanied by the progressive deposition of beta amyloid (Abeta) in both senile plaques and cerebral blood vessels, loss of central neurons, and vessel damage. Cerebral hypoperfusion is one of the major clinical features in AD and likely plays a critical role in its pathogenesis. In addition to its major roles in angiogenesis, vascular endothelial growth factor (VEGF) has neurotrophic and neuroprotective effects. VEGF is an ischemia inducible factor and increased expression of VEGF often occurs in AD. Although the presence of VEGF immunoreactivity in the AD brain has been described previously, the direct interaction of VEGF with Abeta has not been established. Here, we show that VEGF is co-localized with Abeta plaques in the brains of patients with AD. In vitro experiments show that VEGF binds to Abeta with high affinity (K(D) approximate to 50 pM). VEGF is co-aggregated with Abeta without any apparent effect on the rate of aggregation, strongly binds to pre-aggregated Abeta, and is very slowly released from the co-aggregated complex. Continuous deposition of VEGF in the amyloid plaques most likely results in deficiency of available VEGF under hypoperfusion and, thus, may contribute to neurodegeneration and vascular dysfunction in the progression of AD. PMID- 15123333 TI - A haplotype of the methylenetetrahydrofolate reductase gene is protective against late-onset Alzheimer's disease. AB - Epidemiological studies have shown that elevated plasma homocysteine (Hcy) levels play an important role in the pathogenesis of Alzheimer's disease (AD). In spite of the evidence that a C677T polymorphism in the methylenetetrahydrofolate reductase (MTHFR) gene elevates plasma Hcy levels, the impact of the C677T polymorphism on the development of AD is controversial. Here, we performed a genetic case-control study in a Japanese population to investigate whether three polymorphisms of the MTHFR gene, C677T (Ala222Val), A1298C (Glu429Ala), and A1793G (Arg594Gln), are associated with the development of late-onset AD (LOAD). In our study, the MTHFR gene had four major regional haplotypes: Haplotype A (677C-1298A-1793G), Haplotype B (677T-1298A-1793G), Haplotype C (677C-1298C 1793G), and Haplotype D (677C-1298C-1793A). The frequency of Haplotype C in LOAD was significantly lower than that in control group. Furthermore, the benefit conferred by the presence of at least one Haplotype C was stronger in LOAD patients who lacked the ApoE 4 allele (OR=0.293; 95% CI=0.115-0.744; P=0.010). The results indicate that Haplotype C of the MTHFR gene is protective against the development of LOAD. PMID- 15123334 TI - Assessment of association between mitochondrial aldehyde dehydrogenase polymorphism and Alzheimer's disease in an older Korean population. AB - BACKGROUND: The mutant allele of mitochondrial aldehyde dehydrogenase (ALDH2(*)2) was found to be associated with Alzheimer's disease (AD) in a Japanese sample, interacting with the apolipoprotein E epsilon 4 allele (Apo E4). OBJECTIVE: In a community Korean population we sought to investigate associations between ALDH2 genotypes and the following outcomes: cognitive impairment, previous cognitive decline, dementia and AD. METHODS: Six hundred ninety community residents aged 65 or over were assessed for demographic characteristics, drinking behaviour, cognitive function, clinical diagnoses of dementia and AD, physical health status, and genotype (ALDH2 and Apo E). RESULTS: There were no significant associations between the ALDH2(*)2 and any cognitive outcome, before or after adjustment for alcohol-related characteristics. These findings were consistent both in the non-drinkers and drinkers. Interaction between ALDH2 and Apo E was only found for one outcome (previous cognitive decline) at borderline levels of significance (P=0.058). CONCLUSIONS: Overall, these findings in a community population did not support a substantial role for ALDH2 genotype in the aetiology of dementia. PMID- 15123335 TI - Hippocampus and entorhinal cortex in mild cognitive impairment and early AD. AB - Magnetic resonance imaging (MRI) has been suggested as a useful tool in early diagnosis of Alzheimer's disease (AD). Based on MRI-derived volumes, we studied the hippocampus and entorhinal cortex (ERC) in 59 controls, 65 individuals with mild cognitive impairment (MCI) and 48 patients with AD. The controls and individuals with MCI were derived from population-based cohorts. Volumes of the hippocampus and ERC were significantly reduced in the following order: control > MCI > AD. Stepwise discriminant function analysis showed that the most efficient overall classification between controls and individuals with MCI subjects was achieved with ERC measurements (65.9%). However, the best overall classification between controls and AD patients (90.7%), and between individuals with MCI and AD patients (82.3%) was achieved with hippocampal volumes. Our results suggest that the ERC atrophy precedes hippocampal atrophy in AD. The ERC volume loss is dominant over the hippocampal volume loss in MCI, whereas more pronounced hippocampal volume loss appears in mild AD. PMID- 15123336 TI - Effects of beta-hydroxybutyrate on cognition in memory-impaired adults. AB - Glucose is the brain's principal energy substrate. In Alzheimer's disease (AD), there appears to be a pathological decrease in the brain's ability to use glucose. Neurobiological evidence suggests that ketone bodies are an effective alternative energy substrate for the brain. Elevation of plasma ketone body levels through an oral dose of medium chain triglycerides (MCTs) may improve cognitive functioning in older adults with memory disorders. On separate days, 20 subjects with AD or mild cognitive impairment consumed a drink containing emulsified MCTs or placebo. Significant increases in levels of the ketone body beta-hydroxybutyrate (beta-OHB) were observed 90 min after treatment (P=0.007) when cognitive tests were administered. beta-OHB elevations were moderated by apolipoprotein E (APOE) genotype (P=0.036). For 4+ subjects, beta-OHB levels continued to rise between the 90 and 120 min blood draws in the treatment condition, while the beta-OHB levels of 4- subjects held constant (P<0.009). On cognitive testing, MCT treatment facilitated performance on the Alzheimer's Disease Assessment Scale-Cognitive Subscale (ADAS-cog) for 4- subjects, but not for 4+ subjects (P=0.04). Higher ketone values were associated with greater improvement in paragraph recall with MCT treatment relative to placebo across all subjects (P=0.02). Additional research is warranted to determine the therapeutic benefits of MCTs for patients with AD and how APOE-4 status may mediate beta-OHB efficacy. PMID- 15123337 TI - Sulindac improves memory and increases NMDA receptor subunits in aged Fischer 344 rats. AB - Inflammatory processes in the central nervous system are thought to contribute to Alzheimer's disease (AD). Chronic administration of nonsteroidal anti inflammatory drugs (NSAIDs) decreases the incidence of Alzheimer's disease. There are very few studies, however, on the cognitive impact of chronic NSAID administration. The N-methyl-d-aspartate (NMDA) receptor is implicated in learning and memory, and age-related decreases in the NMDA NR2B subunit correlate with memory deficits. Sulindac, an NSAID that is a nonselective cyclooxygenase (COX) inhibitor was chronically administered to aged Fischer 344 rats for 2 months. Sulindac, but not its non-COX active metabolite, attenuated age-related deficits in learning and memory as assessed in the radial arm water maze and contextual fear conditioning tasks. Sulindac treatment also attenuated an age related decrease in the NR1 and NR2B NMDA receptor subunits and prevented an age related increase in the pro-inflammatory cytokine, interleukin 1beta (IL-1beta), in the hippocampus. These findings support the inflammation hypothesis of aging and have important implications for potential cognitive enhancing effects of NSAIDs in the elderly. PMID- 15123338 TI - Long-term dietary restriction causes negative effects on cognitive functions in rats. AB - Long-term dietary restriction is reported to increase life span and improve age related cognitive deficits. The present study shows that the restriction increases the life span of rats but decreases their cognitive ability. Thirty-two rats were divided into restricted and ad lib feeding groups at 2.5 months of age. The restricted rats were kept at a weight of 280g. The restricted rats were poor in performing the Morris water maze task at 7-12 months. At 17-18 months, they were poor in performing the delayed matching-to-place task. At 24-27 months, the surviving 13 restricted and 5 ad lib rats performed the spatial discrimination task. The restricted rats were also poor in performing this task. Injection of glucose prior to the discrimination task improved their performance to the level of the ad lib rats. These results suggest that dietary restriction is beneficial for longevity but has negative effects on the performance of cognitive tasks, and that the cause of the negative effects may be a reduced availability of glucose in the food-restricted aged rats. PMID- 15123339 TI - Impact of age and caloric restriction on neurogenesis in the dentate gyrus of C57BL/6 mice. AB - Age-related changes in neurogenesis and its modulation by caloric restriction (CR) were studied in C57BL/6 mice. To this end, bromodeoxyuridine (BrdU) labeling was used to assess neuronal and glial precursor proliferation and survival in the granular cell layer (GCL) and the hilus of the dentate gyrus of 2-, 12-, 18-, and 24-month-old mice. For both regions, we found an age-dependent decrease in proliferation but not in survival of newborn cells. Interestingly, the reduction in proliferation occurred between 2 and 18 months of age with no additional decline between 18- and 24-month-old mice. Phenotyping of the newborn cells revealed a decrease in the neuron fraction in the GCL between 2 and 12 months of age but not thereafter. The majority of BrdU cells in the hilus colocalized with astrocytic but none with neuronal markers. CR from 3 to 11 months of age had no effect on neurogenesis in the GCL, but had a survival-promoting effect on newly generated glial cells in the hilus of the dentate gyrus. In conclusion, C57BL/6 mice reveal a substantial reduction in neurogenesis in the dentate gyrus until late adulthood with no further decline with aging. Long-term CR does not counteract this age-related decline in neurogenesis but promotes survival of hilar glial cells. PMID- 15123340 TI - BDNF is necessary for maintenance of noradrenergic innervations in the aged rat brain. AB - In the axon terminals of the locus coeruleus (LC) neurons, a high level of axonal branching was occurred in the middle-aged brain, and the increased branching was maintained in the aged brain. In the present study, we hypothesized that neurotrophic support is necessary for the morphological age-related changes seen in the noradrenergic innervations from the LC to frontal cortex. Through immunohistochemical and quantitative image analyses, we examined the age dependent effects of brain-derived neurotrophic factor (BDNF) on the noradrenergic axon terminals in the frontal cortex of F344 rats. We continuously infused BDNF into the frontal cortex of young (6-months-old), middle-aged (13 months-old), or aged (25-months-old) rats. Exogenous BDNF infusion caused a marked increase in the density of noradrenergic axons in the aged brain, but no trophic action of BDNF was observed in the young and middle-aged brain. Neutralization of endogenous BDNF with a specific function-blocking antibody to BDNF led to a reduction in noradrenergic axons in the frontal cortex of 19-month old rats. The present results suggest that BDNF is not involved in the augmentation of noradrenergic innervations in the aging brain, but it is necessary for the maintenance of noradrenergic innervations in the aged brain. PMID- 15123341 TI - Mitochondrial polarisation status and [Ca2+]i signalling in rat cerebellar granule neurones aged in vitro. AB - Mitochondrial membrane potential is a major factor that controls, ultimately, the cellular energy supply. By use of a mitochondrial membrane potential dye (rhodamine 123, R123) and image analysis we show that during long-term (>3 weeks) culture of primary neurones (cerebellar granule neurones) there is a gradual and time-dependent depolarisation of neuronal mitochondria. This process was demonstrated by analysing the changes in the heterogeneity of the cytosolic rhodamine 123 fluorescent signal as a function of the age in culture and by measuring the amplitude of the rhodamine 123 fluorescence evoked by the addition of a mitochondrial protonophore (FCCP). The relationship between cytosolic [Ca(2+)](i) and mitochondrial membrane potential was assessed by recording both parameters simultaneously, in neurones loaded with fura-2 and rhodamine 123. Neuronal stimulation (KCl-evoked depolarisation) induced a mitochondrial depolarisation response resulting from the entry of cytosolic Ca(2+) into mitochondria. In young cultures (10 DIV), the mitochondrial membrane potential recovered fully within 30s from the start of the stimulation, despite the continuous presence of the depolarisation stimulus and the maintained cytosolic [Ca(2+)](i) signal. In contrast, in older neurones (DIV 22), the mitochondrial response was of smaller amplitude and displayed a much longer repolarization period. Also, in these older neurones, the threshold [Ca(2+)](i) level required for the initiation of the mitochondrial depolarisation response was increased by 50%. Thus, the present results indicate that neuronal maturation and ageing in conditions of long-term in vitro culture determine significant changes in the mitochondrial polarisation status that are manifest both in resting conditions and during stimulation and could explain some of the reported changes in neuronal homeostasis in long-term neuronal cultures. PMID- 15123342 TI - Prominent decline of newborn cell proliferation, differentiation, and apoptosis in the aging dentate gyrus, in absence of an age-related hypothalamus-pituitary adrenal axis activation. AB - Neurogenesis and apoptosis in the hippocampal dentate gyrus (DG) occur during development and adulthood. However, little is known about how these two processes relate to each other during aging. In this study, we examined apoptosis, proliferation, migration, and survival of newborn cells in the young (2 weeks), young-adult (6 weeks), middle-aged (12 months), and old (24 months) rat DG. We also measured dentate volume and cell numbers, along with basal corticosterone and stress response parameters. We show that new cell proliferation and apoptosis slow down profoundly over this time period. Moreover, migration and differentiation into a neuronal or glial phenotype was strongly reduced from 6 weeks of age onwards; it was hardly present in middle-aged and old rats as confirmed by confocal analysis. Surprisingly, we found no correlation between cell birth and corticosterone levels or stress response parameters in any age group. PMID- 15123343 TI - Aging, sexual dimorphism, and hemispheric asymmetry of the cerebral cortex: replicability of regional differences in volume. AB - We examined age-, sex-, and hemisphere-related differences in the cerebral cortex. Volumes of the cerebral hemispheres and 13 regions of interest (ROIs) were measured on magnetic resonance images of 200 healthy adults. The strength of association between age and volume differed across ROIs. The lateral prefrontal cortex exhibited the greatest age-related differences, whereas significantly weaker associations were observed in the prefrontal white matter, sensory-motor, and visual association regions. The hippocampal shrinkage was significant in people in their mid-fifties. The primary visual, anterior cingulate, the inferior parietal cortices, and the parietal white matter showed no age-related differences. The pattern of age-related regional differences replicated the findings previously obtained on an independent sample drawn from the same population. Men evidenced larger volumes in all ROIs except the inferior parietal lobule, even after sexual dimorphism in body size was statistically controlled. In some regions (hippocampus and fusiform gyrus) men exhibited steeper negative age-related trends than women. Although a typical pattern of global hemispheric asymmetry was observed, the direction and magnitude of regional volumetric asymmetry was as inconsistent as in the previous reports. Thus, a pattern of age related shrinkage suggesting increased vulnerability of the lateral prefrontal cortex to aging appears stable and replicable, whereas little consistency exists in sex-related and hemispheric differences in regional cortical volumes. PMID- 15123344 TI - High prevalence of thorn-shaped astrocytes in the aged human medial temporal lobe. AB - Thorn-shaped astrocytes (TSA) are glial fibrillary tangles that contain abnormally phosphorylated and aggregated microtubule-associated tau protein. The present study examines the prevalence of TSA in the human medial temporal lobe of 100 autopsy brains aged 42-97 years (mean age: 65 years). Serial brain sections were cut at 100 microm and stained using phosphorylation-dependent anti-tau antibodies (AT8, PHF-1, TG3, Alz-50) and silver staining methods for neurofibrillary changes and beta-amyloid deposits. TSA preferentially were distributed in periventricular, subependymal, and subpial areas of the mediobasal temporal lobe (MTL). Double-labeling with AT8 and anti-GFAP antibodies demonstrated that the abnormal tau protein was deposited in astroglial cell bodies and in proximal and distal astroglial processes. A pronounced inter individual variation was noted in the density of AT8-positive TSA, thereby allowing distinction of mild, moderate, and severe involvement. TSA were absent in individuals younger than 60 years. A significant increase in the prevalence of TSA was noted with advancing age. In the age-range of 75-98 years TSA were found in approximately 50% of all individuals. The development of TSA was not correlated with the severity of Alzheimer-related cortical pathology. In summary, this study suggests that TSA is a distinct form of glial tau pathology that occurs with a high frequency in elderly individuals. PMID- 15123345 TI - Aging, spatial learning, and total synapse number in the rat CA1 stratum radiatum. AB - The aim of this study was to determine whether spatial learning deficits in aged rats are associated with a loss of hippocampal synapses. The Morris water maze task was used to assess the spatial learning capacity of young and aged rats and to attribute aged animals to learning-impaired and learning-unimpaired groups. The number of axospinous synapses in the entire volume of the CA1 stratum radiatum was estimated with unbiased stereological techniques. The results show that the total number of all axospinous synapses and of their perforated and nonperforated subtypes remains constant in the CA1 stratum radiatum of aged learning-impaired rats as compared to aged learning-unimpaired rats and to young adults. Thus, neither age-related deficits in spatial learning nor advanced chronological age are associated with a loss of axospinous synapses from the rat CA1 stratum radiatum. PMID- 15123347 TI - Mechanisms of HCV reinfection and allograft damage after liver transplantation. PMID- 15123348 TI - Susceptibility to primary sclerosing cholangitis is associated with polymorphisms of intercellular adhesion molecule-1. AB - BACKGROUND/AIMS: Intercellular adhesion molecule-1 (ICAM-1, CD54) gene polymorphisms have been implicated in the susceptibility to a range of inflammatory diseases, including inflammatory bowel disease (IBD). Primary sclerosing cholangitis (PSC) is an immune-mediated chronic cholestatic liver disease associated with IBD. ICAM-1 is expressed on proliferating bile ducts and interlobular bile ducts in late stage PSC and serum levels of soluble intercellular adhesion molecules are increased in PSC. The aim of this study was to analyse ICAM-1 gene polymorphisms in PSC patients. METHODS: In this study, 104 patients with PSC and 213 healthy controls were recruited from Oxfordshire Caucasians. PCR with sequence-specific primers (PCR-SSP) was used to detect both ICAM-1 biallelic polymorphisms G241R and K469E. The results were controlled for the HLA haplotypes associated with PSC. RESULTS: The E/E frequency of K469E in PSC was 12% (12/104), significantly lower than that in controls (24%, 51/213;P = 0.009; Pc = 0.02; OR, 0.41). The occurrence of the haplotype G241-E469/G241-E469 in PSC was 4% (4/104), significantly lower than the control group (13%, 28/213; P = 0.01; Pc = 0.04; OR, 0.26). There was no difference between PSC and control groups in the frequencies of the genotype R241G or in allele frequencies of K469E. CONCLUSIONS: The E469E homozygote status for ICAM-1 is associated with protection against PSC. PMID- 15123349 TI - Primary biliary cirrhosis: an infectious disease caused by Chlamydia pneumoniae? AB - BACKGROUND/AIMS: The etiology and pathogenesis of primary biliary cirrhosis (PBC) remain elusive. Both an infectious etiology and molecular mimicry have been implicated. The aim is to study the prevalence of Chlamydial antigens and RNA in the liver tissue of patients with PBC. METHODS: We compared the prevalence of Chlamydial antigen and RNA in 25 explants with PBC who underwent orthotopic liver transplantation with 105 explanted livers from other chronic liver disease. We also studied 14 liver biopsies from patients with early stages of PBC. Donor livers were also studied. RESULTS: In all 39 patients with PBC, Chlamydia pneumoniae antigens were present but not Chlamydia trachomatis, and only 9/105 (8.5%) of patients in the other categories were positive (P<0.01) for C. pneumoniae. Eight explanted PBC livers were tested for C. pneumoniae 16S RNA by in situ hybridization and were positive. CONCLUSIONS: The presence of C. pneumoniae antigen and RNA in liver tissue of patients with PBC suggests that C. pneumoniae antigen may trigger an immune response based on molecular mimicry. PMID- 15123350 TI - Experimental cholestatic liver disease through bile-duct ligation in rats results in skeletal fragility and impaired osteoblastogenesis. AB - BACKGROUND/AIMS: Patients with cholestatic liver disease have 'low-turnover' osteoporosis. Since we reported that bile-duct ligated (BDL) rats develop bone disease with low bone formation and mass, we examined whether their reduced bone mass results in skeletal fragility, and whether the reduction in osteoprogenitor cells could explain the depressed bone formation. METHODS: Four-week-old rats were pair-fed and subjected to BDL or sham surgery. After 4 weeks, ex vivo bone marrow stromal cell cultures were used to estimate the number of osteoprogenitors and tibial strength was measured by mechanical testing. The serum levels of albumin, bilirubin, alanine amino-transferase (ALT), alkaline phosphatase (ALP) and nitrite were measured. RESULTS: BDL rats had elevated levels of bilirubin, ALT, ALP and nitrite. Tibiae of BDL rats were weaker than those of sham rats, exhibiting lower maximal force (-34%) and stiffness (-37%). The number of mineralized bone-like nodules in cultures from BDL rats was 65% lower than that in cultures from sham-operated rats, attesting to a diminished number of osteoprogenitors. CONCLUSIONS: Skeletal fragility diminished osteoprogenitor pool and elevated plasma levels of nitrite are three additional characteristics of the bone disease that develops in BDL rats, thus increasing the validity of this animal model as representing the human bone disease in patients with cholestatic liver disease. PMID- 15123351 TI - Hepatocyte growth factor-induced proliferation of hepatic stem-like cells depends on activation of NF-kappaB. AB - BACKGROUND/AIMS: Hepatocyte growth factor (HGF) regulates proliferation of hepatic stem cells. Transcription factor nuclear factor kappa B (NF-kappaB) has been demonstrated as a key mediator for cell growth regulation. We investigated the role of NF-kappaB in HGF-mediated cellular proliferation responses in a rat liver-derived hepatic stem-like cell line WB-F344. METHODS: Cell proliferation was determined by incorporation of [3H]thymidine. Phosphorylation of ERK1/2, p38 MAPK, Akt and IkappaBalpha by HGF stimulation was detected by Western blotting. NF-kappaB activation was determined by electrophoretic mobility shift assay and NF-kappaB-mediated SEAP reporter assay. NF-kappaB activation was inhibited by treatment with an IkappaBalpha dominant-negative vector or inhibitor BAY-11-7082. RESULTS: We found that stimulation of WB-F344 cells with HGF promoted cell proliferation and effectively protected WB-F344 cells from apoptosis induced by TNF-alpha. We also observed activation of ERK1/2, p38 MAPK, Akt and NF-kappaB signaling pathways by HGF in WB-F344 cells. HGF-induced cell proliferation was partly blocked by pre-treatment of the cells with inhibitors against MEK1 or p38 MAPK, and completely blocked using an inhibitor for NF-kappaB activity. Furthermore, it was demonstrated that IkappaB mutant that suppressed NF-kappaB activity completely blocked HGF-induced cell proliferation. CONCLUSIONS: NF kappaB activity is required for HGF-induced proliferation in hepatic stem-like cell line WB-F344, and this activity requires ERK1/2 and p38 MAPK pathways. PMID- 15123352 TI - Activin A augments vascular endothelial growth factor activity in promoting branching tubulogenesis in hepatic sinusoidal endothelial cells. AB - BACKGROUND/AIMS: The production of activin A is markedly up-regulated in hepatocytes after partial hepatectomy. This factor tonically inhibits growth of hepatocytes but little is known about its effect on sinusoidal endothelial cells (SEC). In the present study, we investigated whether or not activin A affects growth and differentiation of SEC. METHODS: Growth and survival of SEC were measured in monolayer culture. Capillary formation was studied using SEC cultured in a collagen gel. RESULTS: SEC could not survive in the absence of vascular endothelial growth factor (VEGF). Activin A had a small effect on prevention of cell death and also enhanced anti-apoptotic action of VEGF. In addition, activin A and VEGF acted synergistically to stimulate cell growth of SEC. In the collagen gel, VEGF induced capillary formation of SEC. Activin A had little effect on branching tubulogenesis by itself but markedly enhanced tubular formation induced by VEGF. Finally, VEGF induced the expression of activin A and activin A increased the expression of VEGF receptors in cultured SEC. CONCLUSIONS: Activin A augments VEGF activity in promoting growth and tubulogenesis of SEC. PMID- 15123353 TI - In vivo real-time microangiography of the liver in mice using synchrotron radiation. AB - BACKGROUND/AIMS: The aim of this study was to visualize hepatic microvessels (less than 100 microm in diameter) in vivo, which could not be visualized by conventional X-ray angiography, by using synchrotron radiation (ultra-bright and monochromatic X-ray). METHODS: Five female Balb/c nu/nu mice were used. To investigate the hepatic microvessels under nearly physiologic conditions, we performed in vivo aortography under anesthesia with 370 mgI/ml nonionic iodine contrast medium using monochromatic 17-keV X-rays generated by a synchrotron. Images were captured with a pixel matrix size of 1024 x 1024 at a rate of 30 pictures/s. The field of view was 7 mm x 7 mm and thus the pixel size was approximately 7 microm. Captured images were evaluated both qualitatively and quantitatively. RESULTS: Small hepatic arterial and portal venous branches of the liver were visualized separately during one sequential aortogram. The minimum diameter of the vessels observed was approximately 20 microm, and the vessels which ran parallel to the hepatic artery were observed and it seemed to be intrahepatic peribiliary arterial plexus. CONCLUSIONS: Our new experimental model would be useful for visualization of changes in the hepatic microcirculation under nearly physiologic conditions. PMID- 15123354 TI - Tetrathiomolybdate in the treatment of acute hepatitis in an animal model for Wilson disease. AB - BACKGROUND/AIMS: Tetrathiomolybdate (TTM) is a potent copper-chelating agent that has been shown to be effective in Wilson disease patients with neurological symptoms. Here, we investigate the potential use of TTM in treating the acute hepatic copper toxicosis in Long-Evans Cinnamon (LEC) rats, an authentic model for Wilson disease. METHODS: After the onset of acute hepatitis, LEC rats were treated once with 10 mg TTM/kg. After 1 and 4 days, parameters of liver toxicity and the subcellular distribution and binding of copper and iron were studied. RESULTS: In 11 out of 12 rats TTM rapidly improved acute hepatitis. Hepatic copper decreased through removal from cytosolic metallothionein and lysosomal metallothionein polymers. The remaining lysosomal copper forms a metallothionein copper-TTM complex. In an almost moribund rat, however, TTM caused severe hepatotoxicity with fatal outcome. CONCLUSIONS: TTM is effective in treating acute hepatitis in LEC rats when applied before the animals become moribund. TTM appears to act by removing the presumable reactive copper associated to lysosomal metallothionein polymers. The remaining lysosomal copper seems to be inactivated by forming a complex with TTM. Moreover, TTM removes copper from cytosolic copper containing metallothionein. As a consequence, metallothionein is degraded and the uptake of copper-metallothionein into the lysosomes and the formation of the metallothionein polymer associated copper is reduced. PMID- 15123355 TI - Inappropriately low angiotensin II generation: a factor determining reduced kidney function and survival in patients with decompensated cirrhosis. AB - BACKGROUND/AIMS: Angiotensin II contributes to the post-glomerular arteriolar vasoconstriction which maintains the glomerular filtration rate (GFR) in renal hypoperfusion. To explore whether depressed angiotensin II generation, due to reduced angiotensinogen production or low angiotensin-converting enzyme (ACE) levels, could impair kidney function in advanced cirrhosis. METHODS: We studied and prospectively followed up 21 diuretic-free ascitic cirrhotic patients, through these determinations: plasma levels of active renin (AR), renin activity (PRA), angiotensin II, ACE and aldosterone; renal clearances of sodium, inulin and para-aminohippurate; antipyrine clearance. Fifteen healthy subjects were also studied. RESULTS: GFR distribution was bimodal, 10 patients had low GFR values (l GFR group) and 11 had normal-GFR values (n-GFR group) (below and above 105 ml/min per 1.73 m(2) body surface area). Antipyrine clearance and Child-Pugh score did not differ in the two patient groups. l-GFR group had higher AR and PRA values, lower ACE levels and a significantly higher AR/Angiotensin II ratio than n-GFR group (all P<0.01). All 21 patients showed increased values of the AR/PRA ratio, i.e. subnormal angiotensinogen levels (P<0.03). The 18-month survival rates of l GFR and n-GFR groups were 20 and 81% (P<0.02). CONCLUSIONS: Low-GFR cirrhotic patients had a worse survival rate associated with more severe contraction of the effective arterial blood volume, higher AR/Angiotensin II ratio and lower ACE levels. PMID- 15123356 TI - High-density lipoproteins reduce the effect of endotoxin on cytokine production and systemic hemodynamics in cirrhotic rats with ascites. AB - BACKGROUND/AIMS: Hypersensitivity to endotoxin is a recognized feature in cirrhosis. High-density lipoproteins (HDL) have a high capacity to inactivate endotoxin. The aim was to determine if HDL reduces the effect of endotoxin on cytokine production and systemic hemodynamics in experimental cirrhosis. METHODS: The study was performed in control and rats with carbon-tetrachloride induced cirrhosis with ascites. Hemodynamic parameters were determined before and after several doses of endotoxin. The effects of 25 microg/kg of endotoxin on the serum concentration of TNFalpha and mean arterial pressure (MAP) were determined after treatment with HDL (80 mg/kg) or saline. RESULTS: Whereas endotoxin decreased MAP only at doses of 100 and 1000 microg/kg in control rats, in cirrhotic rats significant hypotension occurred at doses of 25, 50, 100 and 1000 microg/kg. Following the administration of endotoxin (25 microg/kg) the serum levels of TNFalpha were 140 times higher in cirrhotic than in control rats (89?835+/-21?090 vs. 625+/-137 pg/ml; P<0.001). Serum TNFalpha was 80% lower in cirrhotic rats pretreated with HDL (18?890+/-5012 pg/ml; P<0.001) than in those pretreated with saline. The administration of endotoxin (25 microg/kg) was associated with a significant lower decrease of MAP in cirrhotic rats pretreated with HDL than in those receiving saline (11.9+/-3.5 vs. 24.7+/-4.3%; P<0.05). CONCLUSIONS: HDL attenuates the increased effect of endotoxin on cytokine production and systemic hemodynamic in cirrhosis. PMID- 15123358 TI - Effects of liver failure on branched-chain alpha-keto acid dehydrogenase complex in rat liver and muscle: comparison between acute and chronic liver failure. AB - BACKGROUND/AIMS: Branched-chain alpha-keto acid dehydrogenase (BCKDH) complex catalyses the committed step in the branched-chain amino acid (BCAA) catabolic pathway. In many cases of liver failure, the serum BCAAs/aromatic amino acids ratio (Fisher's ratio) decreases, and BCAAs have been administered to patients with liver failure to correct this ratio. We conducted an animal study to examine whether the effects on hepatic BCKDH complex differ between acute liver failure (ALF) and chronic liver failure (CLF). METHODS: ALF and CLF was induced in rats by a single high-dose injection and 21 weeks of repeated low-dose injections of carbon tetrachloride, respectively. Plasma BCAA and branched-chain alpha-keto acid (BCKA) levels, and activities and protein amounts of hepatic BCKDH complex and kinase were measured. RESULTS: ALF was characterized by elevated plasma BCAA and BCKA levels and decreased hepatic BCKDH activity. CLF was characterized by decreased plasma BCAA and BCKA levels and increased hepatic BCKDH activity. This increase in BCKDH activity in CLF was associated with the decreased BCKDH kinase, which is responsible for the BCKDH inactivation. CONCLUSIONS: The results obtained in the present study suggest that BCAA catabolism is suppressed in ALF and increased in CLF. PMID- 15123357 TI - Connective tissue growth factor induces c-fos gene activation and cell proliferation through p44/42 MAP kinase in primary rat hepatic stellate cells. AB - BACKGROUND/AIMS: Connective tissue growth factor (CCN2) is expressed during activation of hepatic stellate cells (HSC) and promotes HSC proliferation, adhesion, and collagen production. The aim of the study was to investigate CCN2 signaling pathways in HSC. METHODS: Primary HSC were obtained by enzymatic perfusion of rat liver. DNA synthesis was evaluated by [(3)H]thymidine incorporation. Phosphorylation of Elk-1, extracellular signal-regulated kinase (ERK1/2) and focal adhesion kinase (FAK) was evaluated by Western blot. Transcriptional factor binding activity was determined by gel mobility shift assay while c-fos promoter and CCN2 promoter activity was evaluated using luciferase reporters. c-fos mRNA expression was evaluated by Northern blot. RESULTS: CCN2 stimulated DNA synthesis and phosphorylation of FAK, Elk-1 and ERK1/2, the latter of which was blocked by heparin. The serum response element binding activity and luciferase reporter activity of the c-fos promoter, together with expression of c-fos, were enhanced by CCN2. CCN2-induced c-fos gene activation, expression and cell proliferation were blocked by inhibiting ERK1/2 with PD98059. CCN2 promoter activity was enhanced by TGF-beta1 or PDGF via a Smad7-dependent pathway. CONCLUSIONS: CCN2-stimulated HSC DNA synthesis is associated with transient induction of c-fos gene activation and expression as well as activation of the ERK1/2 signal pathway. PMID- 15123359 TI - Induction of Mx-2 in rat liver by toxic injury. AB - BACKGROUND/AIMS: Mx proteins are supposed to be strictly regulated by viruses or interferon-alpha (IFN-alpha). We used a non-viral model of acute liver injury to study Mx expression. METHODS: We induced toxic liver injury by CCl(4), and studied the expression of IFN-alpha, IFN-gamma, and IFN-inducible antiviral genes (Mx-2; 2'-5' oligoadenylate synthetase, 2-5 A; double-stranded RNA-activated protein kinase, PKR). RESULTS: Similar to 2-5 A and PKR, Mx-2 gene expression was biphasically induced after CCl(4) administration with a maximum at 24 h, and a second peak at 72 h. On protein level, Mx-2 only was up-regulated. IFN-alpha remained constant for the first 24 h while IFN-gamma peaked at 6 h. Thereafter, IFN-alpha increased to a maximum at 72 h while IFN-gamma decreased to 77+/-4%. Small monocyte-like liver macrophages, but not large macrophages, expressed Mx-2 constitutively. In vitro, IFN-alpha but not IFN-gamma induced Mx-2 in different liver cell populations. IFN-gamma, instead, reduced the susceptibility of liver macrophages to the actions of IFN-alpha. CONCLUSIONS: Our data suggest that Mx expression does not invariably result from the presence of a viral particle or IFN-alpha synthesis but may represent an innate defensive armamentarium that may be up-regulated without antigen specificity upon liver injury. PMID- 15123360 TI - Toxicity of low dose azathioprine and 6-mercaptopurine in rat hepatocytes. Roles of xanthine oxidase and mitochondrial injury. AB - BACKGROUND/AIMS: To study effects of pharmacologic concentrations of azathioprine and 6-mercaptopurine (6-MP) on rat hepatocytes. METHODS: Hepatocytes cultured on matrigel were incubated with azathioprine or 6-MP; effects of putative protective agents were studied. Viability (LDH leakage), reduced (GSH) and oxidized glutathione (GSSG), mitochondrial (mt) GSH, ATP and ultrastructural changes were determined. RESULTS: Azathioprine and 6-MP (0.5-5 micromol/l) reduced viability 5 34% at day 1 and 42-92% by day 4. Allopurinol (20 microM) (xanthine oxidase inhibitor) and 2 mM Trolox (vitamin E analog) together provided near complete protection. During culture with azathioprine, GSSG increased before cell death and there was a disproportionate reduction of mtGSH and ATP, together with ultrastructural abnormalities in mitochondria. All changes were prevented by allopurinol and trolox. Discontinuation of 1 micromol/l azathioprine restored ATP levels and arrested cell injury, while culture in glucose-enriched media augmented ATP levels and ameliorated cell death. CONCLUSIONS: Clinically relevant concentrations of azathioprine and 6-MP are toxic to rat hepatocyte cultures by a mechanism that involves oxidative stress, mitochondrial injury and ATP depletion. This can lead to irreversible de-energization and cell death by oncosis (necrosis). PMID- 15123361 TI - Mechanism of liver regeneration after partial hepatectomy using mouse cDNA microarray. AB - BACKGROUND/AIMS: The liver has the capacity to regenerate after partial hepatectomy. In order to clarify the mechanism of liver regeneration, we observed the initial stage, especially the mechanism of gene expression during progress from G0 to S phase (0-24 h), and attempted to identify new genes controlling progress to the S phase. METHODS: We applied large-scale gene expression analysis with complementary DNA microarrays in mouse hepatectomy models to clarify the mechanism of liver regeneration after partial hepatectomy. RESULTS: As a result, 23 new immediate-early gene candidates such as interleukin-1 receptor associated kinase-1 and karyopherin alpha-1, which are involved in transportation within the nucleus, were discovered. Candidates for new genes concerned with the progress to the S phase were discovered: inhibitor of DNA binding 2 (ID2) and inhibitor of DNA binding 3 (ID3), both new liver regeneration factors that promoted progress to the S phase, and GADD45 gamma (growth arrest and DNA-damage-inducible protein) as a factor inhibiting that process. CONCLUSIONS: The above results not only suggest the importance of NFkappaB in the initial stage of liver regeneration but also points to the orderly maintenance of the proliferation of the cells in liver regeneration. PMID- 15123362 TI - Rising incidence of intrahepatic cholangiocarcinoma in the United States: a true increase? AB - BACKGROUND/AIMS: The incidence of intrahepatic cholangiocarcinoma (ICC) has been reported to be increasing in the USA. The aim of this study is to examine whether this is a true increase or a reflection of improved detection or reclassification. METHODS: Using data from the Surveillance Epidemiology and End Results (SEER) program, incidence rates for ICC between 1975 and 1999 were calculated. We also calculated the proportions of cases with each tumor stage, microscopically confirmed cases, and the survival rates. RESULTS: A total of 2864 patients with ICC were identified. The incidence of ICC increased by 165% during the study period. Most of this increase occurred after 1985. There were no significant changes in the proportion of patients with unstaged cancer, localized cancer, microscopic confirmation, or with tumor size <5 cm during the period of the most significant increase. The 1-year survival rate increased significantly from 15.8% in 1975-1979 to 26.3% in 1995-1999, while 5-year survival rate remained essentially the same (2.6 vs. 3.5%). CONCLUSIONS: The incidence of ICC continues to rise in the USA. The stable proportions over time of patients with early stage disease, unstaged disease, tumor size <5 cm, and microscopic confirmation suggest a true increase of ICC. PMID- 15123363 TI - Combination therapy with amantadine and interferon in naive patients with chronic hepatitis C: meta-analysis of individual patient data from six clinical trials. AB - BACKGROUND/AIMS: In chronic hepatitis C, clinical trials evaluating the efficacy of amantadine (AMA) and interferon (INF) compared to INF monotherapy, have produced conflicting results. We performed a meta-analysis of the individual patient's data from previous studies. METHODS: Nine hundred and seventy-two patients from six European centres were evaluated by means of individual patient meta-analysis, using mixed models with centres and the centre-treatment interaction fitted as random variables. RESULTS: At the end of therapy, virological responses were 38.5% (95% CI 34.1-42.8) after INF and AMA, and 29.5% (95% CI 25.5-33.6) after INF alone (P = 0.003). Sustained response occurred in 111 (23.1%; 95% CI 19.3-20.2) and 85 patients (17.3%; 95% CI 14.0-20.7), respectively (P = 0.03). Even accounting for the centre effect, therapy with AMA and INF was more effective than IFN alone (P = 0.029). When genotypes and viraemia levels were combined, the response rate after combination therapy doubled that observed with IFN alone in all subgroups, except those with low viraemia and genotypes 2 or 3. CONCLUSIONS: In chronic hepatitis C, therapy with AMA and INF is effective and may be an alternative to INF and ribavirin in patients who cannot tolerate ribavirin. PMID- 15123365 TI - Complementary and alternative therapies in the treatment of chronic hepatitis C: a systematic review. AB - BACKGROUND/AIMS: Hepatitis C is an escalating global health problem. The recommended treatment regimen is associated with considerable expense, adverse effects and poor efficacy in some patients. Complementary therapies are widely promoted for and used by patients with hepatitis C. The aim is to systematically assess the efficacy of complementary therapies in treating chronic hepatitis C. METHODS: Systematic searches were conducted in six databases, reference lists of all papers were checked for further relevant publications and information was requested from experts. No language restrictions were imposed. RESULTS: Twenty seven eligible randomised clinical trials were located involving herbal products and supplements. No randomised clinical trials were identified for any other complementary therapy. In 14 of the trials, patients received interferon-alpha in combination with the complementary therapy. Less than half the trials (11/27) were of good methodological quality. Compared with the control group, significant improvements in virological and/or biochemical response were seen in trials of vitamin E, thymic extract, zinc, traditional Chinese medicine, Glycyrrhiza glabra and oxymatrine. CONCLUSIONS: We identified several promising complementary therapies, although extrapolation of the results is difficult due to methodological limitations. More research is warranted to establish the role of these and other therapies in the treatment of hepatitis C. PMID- 15123364 TI - The impact of steatosis on disease progression and early and sustained treatment response in chronic hepatitis C patients. AB - BACKGROUND/AIMS: Questions remain regarding the etiology of steatosis in hepatitis C, and its impact on disease progression and treatment outcomes. METHODS: We evaluated liver biopsies from 574 patients with chronic hepatitis C from a single center. RESULTS: Severity of steatosis was associated with body mass index, HCV genotype 3 infection, age, and duration of infection (P1 mm) needles (CN). As fine needles (FN) do not require anaesthesia, are used over a wider range of coagulation values and allow multiple passes, we compared the diagnostic yield of FN vs. CN biopsies. METHODS: Paired samples obtained with FN (0.8 mm) and CN (1.2 mm) on 149 consecutive outpatients from a tertiary care institution were evaluated prospectively. Histologic variables were quantitatively scored. Sensitivity, specificity, positive predictive value, negative predictive value and positive likelihood ratio were calculated as measures of diagnostic ability assuming CN as reference. RESULTS: FN biopsy was adequate in 83 cases, CN in 140 cases (P<0.001). Considering the 83 paired adequate specimens, the best sensitivity of FN vs. CN was for portal inflammation (0.95%) and the worst for cirrhosis (0.33%). Overall discriminant ability of FN was unsatisfactory and histologic variables were systematically underscored. Tolerability was good for both procedures. CONCLUSIONS: The advantages of FN biopsy are lost on its inferior diagnostic performance. Its use in diffuse liver diseases should be restricted to early non-fibrotic lesions. PMID- 15123367 TI - Mutations in the basic core promoter region of hepatitis B virus. Relationship with precore variants and HBV genotypes in a Spanish population of HBV carriers. AB - BACKGROUND/AIMS: To determine the prevalence and significance of hepatitis B virus (HBV) basic core promoter (BCP) mutations and to establish their relationship with precore (preC) mutations, HBV genotypes and HBV-DNA levels. METHODS: BCP and preC mutations and genotypes were determined by sequencing. RESULTS: Genomic analysis was performed in 129 (71%) of 182 patients. BCP mutations were detected in 83% of 18 HBeAg-negative (e-) chronic hepatitis B (CHB) patients with fluctuating ALT levels, and in 76% of 58 e- CHB with elevated ALT. The prevalence was lower and similar, 55% in 30 HBeAg-positive CHB (e+ CHB) with elevated ALT and in 23 e- inactive carriers. Frequency of preC mutations was higher in e- CHB (80%) than in e- inactive carriers (65%). Among e- CHB, patients with elevated ALT and preC mutations at nt 1896 showed highest HBV-DNA, regardless of BCP mutations. BCP mutations were similar in genotypes A and D, while preC mutations were most common in genotype D (82 vs. 40%). Simultaneous presence of the main BCP (1762, 1764) and preC (1896, 1899) mutations was associated with the degree of histological injury. CONCLUSIONS: Combined BCP and preC mutational and genotype analysis provides clinically relevant information in the study of HBV infection. PMID- 15123368 TI - Non-sequencing molecular approaches to identify preS2-defective hepatitis B virus variants proved to be associated with severe liver diseases. AB - BACKGROUND/AIMS: PreS2-defective hepatitis B virus (HBV) variants may emerge during chronic HBV infection. These variants carry mutation(s) at the ATG-start codon and/or in-frame deletion into the preS2 genomic region and are commonly detected by sequencing analyses. We evaluated the prevalence of these variants in a large series of chronic HBV infected patients through non-sequencing molecular approaches. METHODS: We examined HBV isolates from 110 HBV carriers: 15 were inactive carriers (IC); 50 had chronic hepatitis (CH); 25 were cirrhotics; 19 had hepatocellular carcinoma (HCC). The entire preS2 genomic region was amplified by PCR technique. The amplicons were processed: (A) through electrophoresis on acrylamide gel to reveal deleted genomes; (B) through electrophoresis on agarose gel after digestion by NlaIII enzyme that cuts the wild ATG-start-codon but not the mutated one. RESULTS: We detected preS2 variants in 56/110 cases (51%). In particular, we found preS2-defective mutants in 2/15 IC, 25/50 CH, 13/26 cirrhotics, and 16/19 HCC. The presence of these variants was thus significantly associated with active infection and liver disease (P<0.002). Moreover, among cases with liver disease preS2-mutants were more prevalent in HCC patients (P<0.02). CONCLUSIONS: Our non-sequencing molecular methods are sensitive and specific, and simplify the identification of all preS2 HBV variant forms. Infection by these variants is significantly associated with active infection and HCC. PMID- 15123369 TI - Hepatitis B virus enhances transduction of human hepatocytes by SV40-based vectors. AB - BACKGROUND/AIMS: Chronic HBV infection, a world-wide epidemic, can lead to chronic hepatitis and eventually to cirrhosis and hepatocellular carcinoma. The liver poses obstacles for many available gene-transfer vectors. SV40-based vectors can transduce human hepatic and hematopoietic cells. We studied the effect of HBV on the transduction - efficiency of human hepatic cells by SV40 - based vectors. METHODS: A SV40-vector carrying the luciferase gene, and wild-type SV40, were used to assess transduction efficiency of human HBV-positive and HBV negative hepatic cells. Transduction efficiency was measured as luciferase activity or by T-antigen staining. To evaluate whether differences in transduction efficiency are due to cell recognition and/or nuclear transport, MHC I receptors were measured by FACS analysis and SV40-DNA was extracted from the nuclei of transduced cells and quantified. RESULTS: Two HBV-positive cell-lines, HepG2.2.2.15 and FLC4-A10II, were transduced significantly more efficiently than their parental HBV-negative cell-lines. Transient transfection of HuH-7 cells with the HBV genome also increased transduction efficiency. The level of MHC-I, the cellular receptor for SV40, was comparable in all the cell-lines studied. However, soon after infection with SV40, the nuclei of HepG2.2.2.15 contained >6 fold more SV40-DNA than HepG2. CONCLUSIONS: HBV increases transduction by SV40 vectors. This is due to enhanced vector entry and/or transport into the nucleus. SV40-vectors appear to have a potential for gene therapy for the treatment of HBV infections. PMID- 15123371 TI - Hepatitis C virus and liver steatosis: when fat is not beautiful. PMID- 15123370 TI - Hepatitis C virus Core Antigen as a predictor of non-response in genotype 1 chronic hepatitis C patients treated with peginterferon alpha-2b plus ribavirin. AB - BACKGROUND/AIMS: Chronic hepatitis C patients infected by genotype 1 are the least responsive to combination therapy and therefore monitoring response is important in identifying non-responders quickly, permitting therapy discontinuation and avoiding side effects and costs. We examined the usefulness of measuring total HCV Core Ag in early treatment with peginterferon alpha-2b and ribavirin in genotype 1 patients in the prediction of response and compared the results with those from HCV RNA quantification. METHODS: Two hundred and sixty eight serum samples from 46 genotype 1 patients receiving combination therapy were examined for HCV Core Ag and quantitative HCV RNA. RESULTS: At baseline, mean HCV RNA and HCV Core Ag concentrations were significantly lower in sustained virologic responders than in non-responders. The negative predictive value of HCV Core Ag testing in predicting non-response at week 12 is 100%, and for a 2 log drop in HCV RNA, using two quantitative tests, it is 88%. CONCLUSIONS: HCV Core Ag determination allows the identification of non-responders with only one test at week 12 and permits stopping therapy in these patients. HCV Core Antigen testing is cheaper and easier to perform than HCV RNA quantification. PMID- 15123372 TI - Monitoring response to antiviral treatment by serum hepatitis C virus core antigen: too early to take shortcuts? PMID- 15123373 TI - Regulation of bile acid synthesis: pathways, nuclear receptors, and mechanisms. PMID- 15123374 TI - Oral glutamine challenge and magnetic resonance spectroscopy in three patients with congenital portosystemic shunts. AB - BACKGROUND/AIMS: Congenital portosystemic shunts are rare abnormalities of liver vasculature that can cause neurological symptoms, probably secondarily to the effects of the metabolism of ammonia in the brain. Our aim was to investigate the relationship between capillary blood ammonia after oral glutamine challenge and magnetic resonance spectroscopy in three patients with congenital portosystemic shunts. METHODS: Neuropsychological tests, oral glutamine challenge and magnetic resonance spectroscopy were performed at baseline and after 6 months of follow-up in three patients with congenital portosystemic shunts. The results were compared to those obtained in a group of six cirrhotic patients with prior episodes of hepatic encephalopathy and healthy controls. RESULTS: Patients with congenital portosystemic shunts exhibited abnormalities of neuropsychological tests, magnetic resonance spectroscopy and a response to the oral glutamine challenge similar to those observed in patients with cirrhosis. The intensity of the rise of brain glutamine was correlated to the area under the curve of ammonia after the oral glutamine challenge (R=0.72). CONCLUSIONS: Neurological manifestations of patients with congenital portosystemic shunts are mediated through similar mechanisms that are involved in the pathogenesis of hepatic encephalopathy. The area under the curve appears to be the better parameter that defines the response to the oral glutamine challenge. PMID- 15123375 TI - Biliary ascariasis. PMID- 15123376 TI - Duplex polymerase chain reaction-restriction fragment length polymorphism assay for rapid detection of HFE mutations-C282Y occurs with a low frequency in Tehran's population. PMID- 15123377 TI - Hepatotoxicity induced by clopidogrel. PMID- 15123378 TI - Hepatitis C virus genotype 1 associated with massive steatosis of the liver and hypo-beta-lipoproteinemia. PMID- 15123379 TI - Instantaneous response to surgically induced hypoxia of hepatocellular carcinoma. PMID- 15123380 TI - Preformed antibodies to biliary epithelial cells and acute rejection of liver allografts. PMID- 15123382 TI - Assessment of the safety of foods derived from genetically modified (GM) crops. AB - This paper provides guidance on how to assess the safety of foods derived from genetically modified crops (GM crops); it summarises conclusions and recommendations of Working Group 1 of the ENTRANSFOOD project. The paper provides an approach for adapting the test strategy to the characteristics of the modified crop and the introduced trait, and assessing potential unintended effects from the genetic modification. The proposed approach to safety assessment starts with the comparison of the new GM crop with a traditional counterpart that is generally accepted as safe based on a history of human food use (the concept of substantial equivalence). This case-focused approach ensures that foods derived from GM crops that have passed this extensive test-regime are as safe and nutritious as currently consumed plant-derived foods. The approach is suitable for current and future GM crops with more complex modifications. First, the paper reviews test methods developed for the risk assessment of chemicals, including food additives and pesticides, discussing which of these methods are suitable for the assessment of recombinant proteins and whole foods. Second, the paper presents a systematic approach to combine test methods for the safety assessment of foods derived from a specific GM crop. Third, the paper provides an overview on developments in this area that may prove of use in the safety assessment of GM crops, and recommendations for research priorities. It is concluded that the combination of existing test methods provides a sound test-regime to assess the safety of GM crops. Advances in our understanding of molecular biology, biochemistry, and nutrition may in future allow further improvement of test methods that will over time render the safety assessment of foods even more effective and informative. PMID- 15123383 TI - Unintended effects and their detection in genetically modified crops. AB - The commercialisation of GM crops in Europe is practically non-existent at the present time. The European Commission has instigated changes to the regulatory process to address the concerns of consumers and member states and to pave the way for removing the current moratorium. With regard to the safety of GM crops and products, the current risk assessment process pays particular attention to potential adverse effects on human and animal health and the environment. This document deals with the concept of unintended effects in GM crops and products, i.e. effects that go beyond that of the original modification and that might impact primarily on health. The document first deals with the potential for unintended effects caused by the processes of transgene insertion (DNA rearrangements) and makes comparisons with genetic recombination events and DNA rearrangements in traditional breeding. The document then focuses on the potential value of evolving "profiling" or "omics" technologies as non-targeted, unbiased approaches, to detect unintended effects. These technologies include metabolomics (parallel analysis of a range of primary and secondary metabolites), proteomics (analysis of polypeptide complement) and transcriptomics (parallel analysis of gene expression). The technologies are described, together with their current limitations. Importantly, the significance of unintended effects on consumer health are discussed and conclusions and recommendations presented on the various approaches outlined. PMID- 15123384 TI - The relevance of gene transfer to the safety of food and feed derived from genetically modified (GM) plants. AB - In 2000, the thematic network ENTRANSFOOD was launched to assess four different topics that are all related to the testing or assessment of food containing or produced from genetically modified organisms (GMOs). Each of the topics was linked to a European Commission (EC)-funded large shared cost action (see http://www.entransfood.com). Since the exchange of genetic information through horizontal (lateral) gene transfer (HGT) might play a more important role, in quantity and quality, than hitherto imagined, a working group dealing with HGT in the context of food and feed safety was established. This working group was linked to the GMOBILITY project (GMOBILITY, 2003) and the results of the deliberations are laid down in this review paper. HGT is reviewed in relation to the potential risks of consuming food or feed derived from transgenic crops. First, the mechanisms for obtaining transgenic crops are described. Next, HGT mechanisms and its possible evolutionary role are described. The use of marker genes is presented in detail as a special case for genes that may pose a risk. Furthermore, the exposure to GMOs and in particular to genetically modified (GM) deoxyribonucleic acid (DNA) is discussed as part of the total risk assessment. The review finishes off with a number of conclusions related to GM food and feed safety. The aim of this paper is to provide a comprehensive overview to assist risk assessors as well as regulators and the general public in understanding the safety issues related to these mechanisms. PMID- 15123385 TI - Detection and traceability of genetically modified organisms in the food production chain. AB - Both labelling and traceability of genetically modified organisms are current issues that are considered in trade and regulation. Currently, labelling of genetically modified foods containing detectable transgenic material is required by EU legislation. A proposed package of legislation would extend this labelling to foods without any traces of transgenics. These new legislations would also impose labelling and a traceability system based on documentation throughout the food and feed manufacture system. The regulatory issues of risk analysis and labelling are currently harmonised by Codex Alimentarius. The implementation and maintenance of the regulations necessitates sampling protocols and analytical methodologies that allow for accurate determination of the content of genetically modified organisms within a food and feed sample. Current methodologies for the analysis of genetically modified organisms are focused on either one of two targets, the transgenic DNA inserted- or the novel protein(s) expressed- in a genetically modified product. For most DNA-based detection methods, the polymerase chain reaction is employed. Items that need consideration in the use of DNA-based detection methods include the specificity, sensitivity, matrix effects, internal reference DNA, availability of external reference materials, hemizygosity versus homozygosity, extrachromosomal DNA, and international harmonisation. For most protein-based methods, enzyme-linked immunosorbent assays with antibodies binding the novel protein are employed. Consideration should be given to the selection of the antigen bound by the antibody, accuracy, validation, and matrix effects. Currently, validation of detection methods for analysis of genetically modified organisms is taking place. In addition, new methodologies are developed, including the use of microarrays, mass spectrometry, and surface plasmon resonance. Challenges for GMO detection include the detection of transgenic material in materials with varying chromosome numbers. The existing and proposed regulatory EU requirements for traceability of genetically modified products fit within a broader tendency towards traceability of foods in general and, commercially, towards products that can be distinguished from each other. Traceability systems document the history of a product and may serve the purpose of both marketing and health protection. In this framework, segregation and identity preservation systems allow for the separation of genetically modified and non-modified products from "farm to fork". Implementation of these systems comes with specific technical requirements for each particular step of the food processing chain. In addition, the feasibility of traceability systems depends on a number of factors, including unique identifiers for each genetically modified product, detection methods, permissible levels of contamination, and financial costs. In conclusion, progress has been achieved in the field of sampling, detection, and traceability of genetically modified products, while some issues remain to be solved. For success, much will depend on the threshold level for adventitious contamination set by legislation. PMID- 15123386 TI - Societal aspects of genetically modified foods. AB - This paper aims to examine some of the reasons behind public controversy associated with the introduction of genetically modified foods in Europe the 1990s. The historical background to the controversy is provided to give context. The issue of public acceptance of genetically modified foods, and indeed the emerging biosciences more generally, is considered in the context of risk perceptions and attitudes, public trust in regulatory institutions, scientists, and industry, and the need to develop communication strategies that explicitly include public concerns rather than exclude them. Increased public participation has been promoted as a way of increasing trust in institutional practices associated with the biosciences, although questions still arise as to how to best utilise the outputs of such exercises in policy development. This issue will become more of a priority as decision-making systems become more transparent and open to public scrutiny. The results are discussed in the context of risk assessment and risk management, and recommendations for future research are made. In particular, it is recommended that new methods are developed in order to integrate public values more efficaciously into risk analysis processes, specifically with respect to the biosciences and to technology implementation in general. PMID- 15123387 TI - Safety assessment, detection and traceability, and societal aspects of genetically modified foods. European Network on Safety Assessment of Genetically Modified Food Crops (ENTRANSFOOD). Concluding remarks. AB - The most important results from the EU-sponsored ENTRANSFOOD Thematic Network project are reviewed, including the design of a detailed step-wise procedure for the risk assessment of foods derived from genetically modified crops based on the latest scientific developments, evaluation of topical risk assessment issues, and the formulation of proposals for improved risk management and public involvement in the risk analysis process. PMID- 15123388 TI - Cerebellum development and schizophrenia: an association study of the human homeogene Engrailed 2. AB - Epidemiological data and family studies in schizophrenia show that genetic factors contribute to the vulnerability to this disorder. The homeogene Engrailed 2 (EN2) is specifically involved in patterning the region that gives rise to the cerebellum and controls the plasticity of midbrain dopaminergic neurons. We carried out an association study for a CA repeat polymorphism located in the 3' region of the homeogene EN2. The subjects consisted of 165 patients with schizophrenia and 97 controls matched for age and ethnicity from a French Caucasian population. We found no significant association of schizophrenia with this bi-nucleotide repeat polymorphism of the EN2 gene. PMID- 15123389 TI - Lack of association between the serotonin transporter promoter gene polymorphism and impulsivity or aggressive behavior among suicide attempters and healthy volunteers. AB - The association between a polymorphism in the promoter area of the serotonin transporter (17q11.1-q12) with impulsivity and history of aggressive behavior was studied in a Spanish general hospital. Subjects comprised 216 suicide attempters (152 women and 64 men) and 223 control blood donors (124 women and 99 men). They were classified as S individuals (s/s or s/l) with low expression of the serotonin transporter, and L individuals (l/l) with high expression. The genotype was not associated with high levels of impulsivity (measured with the Barratt Impulsiveness Scale) or history of aggressive behavior (measured with the Brown Goodwin scale). This lack of association did not appear to be explained by lack of statistical power. High scores on the Barratt Impulsiveness Scale (BIS) and the Brown-Goodwin Aggressive Behavior Scale were associated with being an attempter, male gender and borderline personality disorder. While our Spanish suicide attempters and published US suicide attempters have similar BIS scores, our Spanish suicide attempters have significantly lower aggressive behavior scores. If cross-cultural differences in aggressive behavior scores are definitively established, country norms for aggressive behavior scales will need to be developed to compare genetic studies in different countries. PMID- 15123390 TI - Assessment of behavioral and cognitive impulsivity: development and validation of the Lifetime History of Impulsive Behaviors Interview. AB - The construction and initial psychometric evaluation of an interview assessment of clinically significant impulsivity (Lifetime History of Impulsive Behaviors; LHIB) is presented. Personality-disordered and control subjects participated by completing self-report measures of depression, anxiety and social desirability, along with self-report and laboratory analogue measures of impulsivity, and finally the LHIB. The LHIB demonstrated good to excellent internal consistency and test-retest reliability. Supporting concurrent construct validity, scores on the LHIB correlated with other self-report measures of impulsivity. Diagnostic group differences were obtained and the LHIB evidenced concurrent validity in its ability to classify subjects by scores. No relationship was obtained between the LHIB and laboratory analogue measures. While evidence of discriminant validity was mixed, these data suggest that the LHIB may be a useful instrument for the assessment of impulsive behavior. PMID- 15123392 TI - Gender differences in depression: the role of personality factors. AB - The goal of the study was to determine the association between gender and the Big Five personality factors, and to identify the role of personality factors in the association between gender and depression among adults in the United States. Data were drawn from the Midlife Development in the United States Survey (N=3032). Multivariate analysis of variance (MANOVA) was used to examine gender differences on the Big Five personality factors (i.e. agreeableness, neuroticism, openness to experience, extraversion, and conscientiousness). Multivariate logistic regression analyses were conducted to examine the relation between gender and depression, and to test whether this association is moderated by neuroticism. Levels of neuroticism, agreeableness, extraversion, and conscientiousness were significantly higher among females than among males; in contrast, level of openness to experience was significantly higher among males. Female gender was associated with increased odds of experiencing depression. Results showed that neuroticism played a significant contributory role in the relationship between being female and major depression, though the role of gender remained statistically significant after adjustment. These data suggest that gender differences in personality factors, specifically neuroticism, may play a key role in the well-documented gender difference in depression. Our findings indicate that neuroticism may moderate the association between female gender and increased risk of depression among adults. These findings require replication using longitudinal data. PMID- 15123391 TI - Repetitive transcranial magnetic stimulation: a putative add-on treatment for major depression in elderly patients. AB - Repetitive transcranial magnetic stimulation (rTMS) is a recent putative treatment for affective disorders. Several studies have demonstrated antidepressant effects of rTMS in younger patients; we aimed to assess its effect in older outpatients with treatment-resistant major depression. Twenty-four outpatients (mean age=62 years, S.D.=12) with major depression were randomized for sham or real stimulation and received 10 daily rTMS sessions (20 Hz, 2-s trains, 28-s intertrain intervals, 100% of motor threshold) in addition to the antidepressant medication. For sham stimulation, the coil was tilted 90 degrees. Depression severity was assessed using the Hamilton Depression Rating Scale, the Beck Depression Inventory, items from the NIMH self-rated symptom scale, and a visual analog depression scale. Mini-Mental Status Examination performance, memory, and executive and attentional functions were measured to control for cognitive side effects. Depression ratings revealed significant antidepressant effects within 2 weeks in both sham and real stimulation groups; however, there were no between-group differences. Treatment with rTMS was safe; adverse events were rare and not more prevalent in either group, and cognitive assessment did not show any deterioration. We were unable to demonstrate any additional antidepressant effects of real stimulation in elderly patients with treatment resistant major depression. Therapeutic effects of rTMS in this clinically challenging patient group remain to be demonstrated. PMID- 15123393 TI - Moderating effects of major depression on patterns of comorbidity in patients with panic disorder. AB - Given the high rate of co-occurring major depression in patients with panic disorder, it is unclear whether patterns of comorbidity in individuals with panic disorder reported in the literature are associated with panic disorder or with the presence of major depression. Subjects were 231 adult subjects with panic disorder and major depression (n=102), panic disorder without comorbid major depression (n=29), major depression without comorbid panic disorder (n=39), and neither panic disorder nor major depression (n=61). Subjects were comprehensively assessed with structured diagnostic interviews that examined psychopathology across the life cycle. Panic disorder, independently of comorbidity with major depression, was significantly associated with comorbid separation anxiety disorder, simple phobia, obsessive-compulsive disorder, generalized anxiety disorder, and agoraphobia. Major depression, independently of comorbidity with panic disorder, was significantly associated with comorbidity with psychoactive substance use disorders and childhood disruptive behavior disorders. Overanxious disorder was associated with both panic disorder and major depression. Major depression has important moderating effects on patterns of comorbidity of panic disorder in referred adults. PMID- 15123394 TI - Somatic symptoms most often associated with depression in an urban hospital medical setting in Japan. AB - Somatic symptoms are known to be the major manifestation in patients with depression. The aim of the present study was to investigate the major somatic and psychiatric symptoms associated with depression in each sex. Patients with a DSM IV diagnosis of depressive disorders (n=335) and comparison patients without depression (n=425) among new outpatients in an urban hospital medical setting were eligible for study. The relationship between the three most distressing subjective symptoms at the first visit, confirmed by the patient's description on a health questionnaire and the admitting physician's interview, and depression was investigated in each sex. Most (77.4%) of the complaints in patients with depression were somatic. In a simple logistic regression analysis, diarrhea, excessive sweating and psychomotor retardation in men, and headache, depressed mood and grief in women were associated with depression. In multiple logistic regression analysis, diarrhea, excessive sweating and weight loss in men, and headache, dysesthesia and grief in women, as well as sleep disturbance, loss of appetite, general fatigue, loss of interest and agitation in both sexes, were statistically significantly associated with depression. Fever in men was also associated with depression by Fisher's exact test. PMID- 15123395 TI - Familial transmission of suicidal ideation and suicide attempts: evidence from a general population sample. AB - The goals of the study were (1) to determine the association between parental and offspring suicidal ideation and suicide attempts among adult offspring in a general community sample, and (2) to examine the extent to which this association can be explained by mediating processes of mental disorders. Data were drawn from the National Comorbidity Survey (n=8098), a representative household sample of adults aged 15-54 in the United States. The relationships between suicidal ideation and suicide attempts among adult offspring and suicidal ideation and suicide attempt in their parents, compared with those in parents not characterized by suicidal ideation or suicide attempts, were calculated using multiple logistic regression analyses. Analyses were adjusted for differences in sociodemographic characteristics and for mental disorders. Results showed that parental suicidal ideation was associated with a significantly increased likelihood of suicidal ideation [OR=1.7 (1.2, 2.5)] and suicide attempt [OR=1.4 (0.9, 2.1)] among offspring. Parental suicide attempt was associated with increased odds of suicidal ideation [OR=2.0 (1.4, 2.9)] and suicide attempt [OR=2.2 (1.4, 3.4)] among offspring. Comorbid mental disorders contributed to the strength of these associations, but with the exception of the link between parental suicidal ideation and offspring suicide attempt, all remained statistically significant even after adjustment. These data provide initial evidence of familial linkages (parent-offspring) of suicidal ideation and behavior among a sample of adults representative of the US population. The data suggest that comorbid mental disorders contribute to these associations but do not completely account for them. The findings are consistent with and extend results from family, clinical, and high-risk studies suggesting that a familial risk of suicidal ideation and suicide behavior occurs in the general population. Implications for prevention and future research are discussed. PMID- 15123396 TI - Is seasonality of suicides stronger in victims with hospital-treated atopic disorders? AB - The aim of the present study was to test whether the seasonal distribution of suicides differed between atopic and non-atopic suicide victims. A cross sectional comparison of the semi-annual and seasonal distribution of suicides was made by using a 13-year database of all suicides (1296 males, 289 females) committed during the years 1988-2000 in the province of Oulu in Northern Finland. During the first half of the year, the proportion of suicides among atopic patients was significantly higher than that linked with non-atopic patients. Of all atopic patients, 72% committed suicide during the first and 28% during the second half of the year. Suicides among victims without any atopic disorders followed a uniform seasonal distribution throughout the year (50 vs. 50%). The exacerbation of an atopic disorder may increase the risk of suicide in spring; something that should be taken note of in clinical work. PMID- 15123397 TI - Factorial invariance of the CES-D in low socioeconomic status African Americans compared with a nationally representative sample. AB - This study examined the factor structure of the Center for Epidemiologic Studies Depression Scale (CES-D) in low socioeconomic status African Americans (n=426). Confirmatory factor analysis indicated that the four factors-(1) depressed affect, (2) positive affect, (3) somatic complaints, and (4) interpersonal problems-of the CES-D scale previously found in the general population were supported in this sample. These results were cross-validated in other nationally representative samples of African-American participants (n=988) and Caucasians (n=666), and the four-factor structure of the CES-D scale replicated in all three groups in this study. An alternative model was also tested with the factors depressed affect and somatic complaints combined as a single factor, a finding often reported in minority groups. Results indicated a significantly poorer fit for the three-factor model compared with the four-factor model for all three groups. In addition, higher loading differences were significantly evident between African-American and Caucasian groups, while higher loading similarities were found between the two African-American groups. These findings provide further evidence of measurement equivalency of the CES-D scale in samples with differential characteristics including race and socioeconomic status. PMID- 15123398 TI - Introduction to risk assessment and serum risk markers for the prevention of coronary heart disease and other potential conditions that impact men's health, part I: what do I tell my patients? AB - This article covers the basic cholesterol profile and a method of assessing the potential risks for CHD for most men. It is hoped that this overview will assist clinicians and patients in raising the awareness of CHD as the leading cause of mortality in men, and help to place the overall risk for mortality into a proper perspective. PMID- 15123399 TI - Introduction to risk assessment and serum risk markers for the prevention of coronary heart disease and other potential conditions that impact men's health, part II: what do I tell my patients? AB - Markers and other potential tests for cardiovascular risk abound, and a more extensive review of these and other markers can be found elsewhere. Cardiovascular health tends to mirror urologic health for several conditions. It is imperative in the urology setting that clinicians discuss the probability of early morbidity and mortality with patients. This should not undermine the importance of prostate cancer and other diseases, but rather place risks in their proper perspective. Patients seem to need this guidance more than ever because life expectancy for most individuals continues to increase, and the potential for comorbidities seems to increase in the average aging patient. It is hoped that this article will prompt new interest in evaluating some risk factors for various urologic conditions. This may move researchers closer to discovering intervention agents that could impact numerous male health conditions favorably. Improving men's health will require a concerted educational and research effort from several health disciplines, a reminder that different medical specialties have much in common, and the role of preventive medicine is to point out similarities as opposed to differences. PMID- 15123400 TI - Prostate cancer and coronary heart disease: correlation or coincidence? AB - A few past clinical and recent case-control studies of statin use, for example, in patients with and without prostate cancer have not demonstrated its potential for reducing or preventing the risk for this disease, and the potential for benefit may have been a confounding coincidence. Data from larger continuing and future studies will be needed to resolve this issue, but the recent data on cholesterol or dyslipidemia and risk increase or reduction with treatment are interesting, especially because of other potential improvements with therapy in nonprostate cancers. In addition, the finding that some available cancer treatments improve some parameters of the lipid profile is fascinating, and some cancer drugs are being used in a specific cardiovascular disease treatment setting to improve outcome. Even if CHD, dyslipidemia, and the treatment of these conditions has no role in preventing prostate cancer or its progression, what has been lost? CVD is still the leading cause of death of men, and a heart-healthy program for the patient concerned about prostate disease would reduce this primary cause of death. Patients would take a step forward in improving all-cause mortality. Recent data from surveys, however, continue to demonstrate that men have an inadequate understanding of cholesterol and heart disease. Crisis creates opportunity, and individuals working in urology have ample reasons not only to discuss the overall benefits of reducing lipid markers, but to improve cholesterol and CHD awareness as much as health professionals working in other fields of medicine. The marriage between general preventive medicine and urology seems to be inevitable, and in the authors' opinion, this merger will provide the foundation for novel research that could affect patients' lives dramatically. PMID- 15123401 TI - The potential role of cyclooxygenase-2 inhibitors and 5alpha-reductase inhibitors in the prevention of urologic conditions. AB - Prevention of urologic cancer is a new field for urology. It focuses attention on men and women who may not be under the regular care of physicians and are without symptoms of disease. Although risk factors (eg, smoking in bladder cancer, family history or African-American ethnicity in prostate cancer, cryptorchid testes in testis cancer, Von Hippel-Lindau disease in renal cell carcinoma) can identify individuals at a greater risk for genitourinary malignancies, most patients have no risk factors except gender and age. Thus, developing public health recommendations that will have a major impact on these diseases will be challenging. Prevention strategies will be held to a higher standard of safety than traditional cancer treatment unless populations at a high risk for disease can be identified. It will be necessary for urologists to monitor the field of disease prevention because of the high frequency of urologic malignancies and the growing elderly population in the United States. Urologists are frequently providers for and confidants of this patient population and must be able to counsel these men and women about the benefits and risks of such prevention strategies. PMID- 15123402 TI - Screening for prostate cancer: current recommendations. AB - Recent studies have questioned the efficacy of PSA as a marker for the early detection of prostate cancer, but techniques are being investigated to improve the sensitivity and specificity of screening. It is hoped that new methods can differentiate between lethal and nonlethal cancers, thereby avoiding lead-time bias. Even with the current limitations of PSA, the combination of stage migration seen with screening, the recent Scandinavian study showing decrease of disease progression following surgical extirpation, and the known mortality in patients presenting with advanced disease help support PSA screening for prostate cancer. It is hoped that prospective, randomized, long-term screening studies, such as the PLCO and ERSCP trials, will show improved survival using the admittedly imperfect PSA marker in prostate cancer screening. PMID- 15123403 TI - Potential pathologic markers for prostate chemoprevention studies. AB - The field of exploring potential biomarkers for prostate cancer prevention continues to expand rapidly. In addition to a limited scope of histologic alterations, mainly PIN, a growing array of promising technologies (eg, computer assisted analysis), and molecules involved in cell-cycle regulations, angiogenesis, and structural and numerical chromosomal alterations are potential candidates for surrogate endpoint biomarkers for prevention trials. Before any these potential candidates are adopted, however, the technical, interpretational, and validation requirements must be satisfied. More important, it is crucial to demonstrate that modulation of the frequency of the biomarker decreases the rate of cancer development. PMID- 15123404 TI - Risk factors for adult renal cell carcinoma. AB - This study confirmed several independent risk and protective factors for RCC identified in the authors' previous study. Protective factors such as oral contraceptive use and moderate alcohol consumption were identified only in women. Tobacco consumption and severe obesity were the main independent risk factors. There were other modifiable risk markers, however, such as occupational exposure, thiazidic drug intake, and urinary tract infections. The associations between risk factors and RCC were weak, even for tobacco, for which the association was weaker than that for lung cancer. The identified risks involve a large proportion of the population, however, and the risk attributable to these types of exposure is high. The authors' recommendations for the prevention of RCC are therefore similar to those for the prevention of cardiovascular disease and cancer, and should be disseminated to the general population. The high-risk groups identified are too large for a specific early-screening program for RCC, but such screening might be appropriate if restricted to selected age groups. PMID- 15123405 TI - Prevention and treatment of erectile dysfunction using lifestyle changes and dietary supplements: what works and what is worthless, part I. AB - Clinicians working in urology should adhere to the same guidelines that are observed in cardiovascular medicine when dealing with a patient with ED. A golden opportunity exists to discuss lifestyle changes with any man with or concerned about ED. Providing heart-healthy recommendations to men with minimal to extensive ED may produce a twofold impact: (1) patients may be able to affect the future extent of their disease, and (2) patients may become healthier overall. Patients following a heart-healthy lifestyle after a diagnosis of ED or to prevent ED should enjoy increased quality or quantity of life. The time is more than ripe for patients to understand that heart health is tantamount to erectile health. PMID- 15123406 TI - Prevention and treatment of erectile dysfunction using lifestyle changes and dietary supplements: what works and what is worthless, part II. AB - It seems naive to believe that some plants or herbs do not contain specific compounds that could benefit patients with ED. Many supplements have not been investigated in a laboratory or clinical research setting before commercial sale, however,which creates a complex situation. If efficacy is or is not demonstrated through adequate research, then the benefit or lack thereof cannot be mentioned on the label. Furthermore, clinicians and the public cannot be made aware of which compounds or supplements are effective because no general standards for sale exist under the current guidelines. Dietary supplements have received a tremendous amount of publicity. The large and growing market for ED treatment seems to have contributed partly to the promotion of numerous supplements and their apparent benefits. Whether these dietary supplements have merit is questionable. Some supplements may produce results opposite to those advertised. Other supplements may be enjoying the benefits of the placebo effect. Because a placebo response of 25% to 50% has been recorded in clinical trials with effective agents, it is understandable that some supplements enjoy financial success despite the limited research espousing their use. If one to two of four individuals or one of three individuals who try a dietary supplement gain some benefit for their ED, the market for these supplements will remain extraordinary. On a larger scale, of 100,000 men who try a supplement, approximately 25,000 to 50,000 will claim some success. The challenge for clinicians is to discuss the placebo response properly and the need for good research before any intervention, especially supplements, can be advocated for general use. Table 2 summarizes some popular ED supplements and general conclusions that can be drawn from clinical investigations. Some dietary supplements may have an active ingredient that benefits patients with certain types of ED. An exciting area of future dietary supplement research is the ability of certain agents to have a synergistic effect with prescription agents for ED, thereby improving response rates in men that have failed approved ED therapy initially, especially with oral agents. Randomized clinical trials are the best method of determining which dietary supplements will become a part of conventional medicine. Therefore, more randomized trials for dietary supplements are needed so that they may have the opportunity to become a part of the mainstream milieu, which means that more funding needs to be made available for ED research. The coming years of research should bring enormous excitement and objectivity to this area of medicine. PMID- 15123407 TI - The impact of obesity in urology. AB - The incidence and progression of urologic diseases, as well as several urologic cancers.depend on many interrelated factors, such as obesity, diet, genetics, environment, age, and the immune system. Obesity is a risk factor for stress urinary incontinence, ED, infertility, and renal calculi. Numerous publications have demonstrated that a high dietary intake of fat increases prostate cancer risk, although the mechanisms are not clear. Although some reports may demonstrate an association between obesity and prostate cancer, it may be hard to establish because, in general, men with obesity have a high-fat diet. Obesity, recurrent urinary tract infections, increased intake of protein and fried foods, and female sex seem to increase the risk of renal cancer. Environmental toxins seem to be the major factors affecting the incidence of bladder cancers. Thus, dietary modification and other public health measures directed at reducing weight may reduce the incidence of urologic illnesses. More studies are necessary to determine the therapeutic effects of weight loss and dietary modification on the incidence and progression of urologic tumors. PMID- 15123408 TI - Lifestyle recommendations to prevent prostate cancer, part I: time to redirect our attention? AB - This article provides a foundation for clinicians willing to provide lifestyle change recommendations for the prevention of prostate cancer. In part II, more general and specific lifestyle recommendations will be provided. It is imperative to provide patients with realistic and practical recommendations that are not only consistent in the medical literature, but will improve overall compliance. PMID- 15123409 TI - Lifestyle recommendations to prevent prostate cancer, part II: time to redirect our attention? AB - This article provides a foundation for men who want to incorporate lifestyle changes to reduce their risk for prostate cancer and, more importantly, impact all-cause mortality. Table 1 summarizes some of these lifestyle changes that can be recommended to patients in most settings. Minimal time is required to suggest these changes, and a copy of Table 1 can be provided as a reminder to patients. Although these recommendations may seem simple, past studies of men have demonstrated that few (less than 5%) adhere to numerous healthy behaviors simultaneously. It seems to be more common to follow one healthy change in excess than to make multiple changes in moderation. This may be the result of past studies focusing on one lifestyle change to affect disease risk; poor compliance; lack of attention, time, or understanding to this detail; or lack of motivation on the part of the health professional and the patient. Clinical trials of combined moderate lifestyle changes, however, demonstrate that the total effort to make healthy lifestyle changes is more important than one or two behavioral changes in affecting cardiovascular markers, cancer, and all-cause mortality. Recommending a pill is an easy answer, but few supplements for prostate-cancer prevention or total mortality reduction can be recommended, and long-term compliance is a concern with any agent. Additionally, the potential for supplements to increase the risk for prostate cancer or interfere with conventional treatment continues to be a concern, and no dietary supplement has come close to matching the reduction in all-cause mortality observed in clinical trials of lifestyle changes. The time seems ripe to redirect our attention regarding lifestyle changes and prostate cancer risk. What is heart-healthy is prostate-healthy, which makes it more likely that any man concerned about the risk for prostate cancer will make healthy lifestyle changes. PMID- 15123410 TI - The impact of osteoporosis in men treated for prostate cancer. AB - Prostate cancer patients are at significant risk for SREs, with up to 50% of androgen-insensitive patients experiencing an SRE at 24 months. The risk increases with the duration and type of cancer treatment. SREs decrease HRQOL, increase the cost of care, and are associated negatively with overall survival. Screening men at greatest risk (slender white men and men with hormone refractory disease or metastatic disease) with BMD measurements, and initiating empiric therapy (vitamin D3, calcium, parenteral estrogens, bisphosphates) may be warranted. PMID- 15123411 TI - Preventing male osteoporosis: prevalence, risks, diagnosis and imaging tests. AB - There is no universal definition of an osteoporotic fracture. Fractures of the vertebra, hip, and forearm generally are considered osteoporotic fractures. There is an increasing recognition, however, that osteoporosis can lead to fractures at other anatomic sites, including the ribs, humerus, tibia, pelvis, and femur. Excluding these types of fractures could underestimate the total cost and impact of osteoporosis. There are a variety of risk factors for osteoporosis and fracture, but several methods can quantify a patient's risk, and these tools are valuable in guiding clinicians in effective intervention. The author hopes this brief review provides clinicians with an introduction and overview of osteoporosis and its risk factors, screening methods, and procedures. The field of urology is constantly evolving, with a growing need for clinicians to become more knowledgeable about preventive medicine. This and other articles should provide a good foundation for clinicians to alter the increasing prevalence of a medical condition that is one of the most preventable causes of morbidity and mortality in men. PMID- 15123412 TI - Understanding treatments for bone loss and bone metastases in patients with prostate cancer: a practical review and guide for the clinician. AB - Prostate cancer patients are at risk for developing bone loss and bone metastases. Clinicians prescribing ADT should appreciate the potential effects of ADT on BMD as well as the morbidity and mortality that can result from osteoporotic fractures. Measures to address the evaluation of patients and when to treat patients with significant bone loss have been discussed. Bisphosphonates effectively prevent loss of BMD in prostate cancer patients. Treatment of prostate cancer patients with established bone metastases with zoledronic acid should be considered strongly based on the results of the Saad study and other studies of patients with bone metastases with other malignancies. Zoledronic acid is approved by the US FDA for use in men with metastatic hormone-refractory prostate cancer and in the European Union for any patient with bone metastases, including prostate cancer patients,because of the beneficial impact of zoledronic acid on skeletal-related events. There is no validated method to determine which patients might benefit most from bisphosphonate therapy in this setting. Many questions about the use of bisphosphonate therapy in men with prostate cancer must be addressed, both in terms of the use in bone loss and bone metastases. These questions include: What is the optimal timing of therapy? Which bisphosphonate is best? What is the best dose and dose schedule? Do bisphosphonates effectively decrease skeletal fracture rates in patients with osteoporosis? How long should patients receive therapy? Are bisphosphonate "holidays" warranted? What are the long-term skeletal and renal toxicities? Is there a role for sequencing bisphosphonate therapy either before or after chemotherapy? Is bisphosphonate therapy synergistic with certain chemotherapy or other bone-targeted therapies? Which patients are the most likely to benefit from bisphosphonate therapy? What are clinically significant endpoints of bisphosphonate trials in patients with metastatic disease? Does inhibiting bone turnover also inhibit formation of bone metastases? Preliminary work in these areas has been completed, but more questions than answers are available. Given the rising costs of health care, it is imperative that these questions be addressed to best use the health care dollar while offering high-risk patients the best available therapy. At present, no data suggest that bisphosphonates should be used routinely to prevent BMD loss in men with normal BMD or to prevent the development of bone metastases in men with biochemical relapse. Continuing trials may give us guidance in the future. PMID- 15123413 TI - Reducing the toxicity associated with the use of radiotherapy in men with localized prostate cancer. AB - The risk for serious complications associated with modern radiotherapy is relatively low. Compared with conventional radiotherapy, 3DCRT and IMRT allow higher doses to be given more safely. The use of IMRT increases the time and effort required by physicians and physicists. Although there is a clear move toward IMRT, 3DCRT is considered standard. The quality assurance procedures for and fundamental questions regarding IMRT are evolving, including the radiobiologic consequences of altered time-dose fractionation and the greater dose heterogeneity in the target. Defining an accurate target volume and routinely correcting for set-up error and organ movement before each treatment promise to reduce the complications associated with EBRT in the next 5 years. Complications following prostate brachytherapy are better understood now than 10 years ago. Dosimetric or patient selection factors that correlate with a higher risk for complications, such as acute retention, strictures, severe prolonged urinary symptoms, fistulas, proctitis, and impotence, often can be accounted for or avoided. Finally, the role of radiation protectors is beginning to be addressed. It is hoped that these advances will eliminate toxicity associated with radiotherapy and increase cure rates. PMID- 15123414 TI - Prevention and management of prostate cancer chemotherapy complications. AB - Prevention and management of the adverse effects of prostate cancer chemotherapy depend on skilled regimen selection, dose adjustment, use of supportive care strategies, and a thorough understanding of the patient- and regimen-related factors that determine the risk for toxicity. Urologists, radiation oncologists, and primary care providers can play an important role before chemotherapy is prescribed by judicious use of treatments that impair bone marrow and other vital organ function. The current role of chemotherapy in prostate cancer is palliative. Successful palliation depends on reducing cancer-related suffering without introducing treatment-related suffering. Thus prevention and management of toxicity is central to the success of chemotherapy in advanced prostate cancer. PMID- 15123417 TI - Recognition of human mitochondrial tRNALeu(UUR) by its cognate leucyl-tRNA synthetase. AB - Accuracy of protein synthesis depends on specific recognition and aminoacylation of tRNAs by their cognate aminoacyl-tRNA synthetases. Rules governing these processes have been established for numerous prokaryotic and eukaryotic cytoplasmic systems, but only limited information is available for human mitochondrial systems. It has been shown that the in vitro transcribed human mitochondrial tRNA(Leu(UUR)) does not fold into the expected cloverleaf, but is however aminoacylated by the human mitochondrial leucyl-tRNA synthetase. Here, the role of the structure of the amino acid acceptor branch and the anticodon branch of tRNA(Leu(UUR)) in recognition by leucyl-tRNA synthetase was investigated. The kinetic parameters for aminoacylation of wild-type and mutant tRNA(Leu(UUR)) transcripts and of native tRNA(Leu(UUR)) were determined. Solution structure probing was performed in the presence or in the absence of leucyl-tRNA synthetase and correlated with the aminoacylation kinetics for each tRNA. Replacement of mismatches in either the anticodon-stem or D-stem that are present in the wild-type tRNA(Leu(UUR)) by G-C base-pairs is sufficient to induce (i) cloverleaf folding, (ii) improved aminoacylation efficiency, and (iii) interactions with the synthetase that are similar to those with the native tRNA(Leu(UUR)). Leucyl-tRNA synthetase contacts tRNA(Leu(UUR)) in the amino acid acceptor stem, the anticodon stem, and the D-loop, which is unprecedented for a leucine aminoacylation system. PMID- 15123416 TI - Internal gene duplication in the evolution of prokaryotic transmembrane proteins. AB - We investigated the evolution of transmembrane (TM) topology by detecting partial sequence repeats in TM protein sequences and analyzing them in detail. A total of 377 sequences that seem to have evolved by internal gene duplication events were found among 38,124 predicted TM protein sequences (except for single-spannings) from 87 prokaryotic genomes. Various types of internal duplication patterns were identified in these sequences. The majority of them are diploid-type (including quasi-diploid-type) duplication in which a primordial protein sequence was duplicated internally to become an extant TM protein with twice as many TM segments as the primordial one, and the remaining ones are partial duplications including triploid-type. The diploid-type repeats are recognized in many 8-tms, 10-tms and 12-tms TM protein sequences, suggesting the diploid-type duplication was a principle mechanism in the evolutionary development of these types of TM proteins. The "positive-inside" rule is satisfied in whole sequences of both 10 tms and 8-tms TM proteins and in both halves of 10-tms proteins while not necessarily in the second half of 8-tms proteins, providing fit examples of "internal divergent topology evolution" likely occurred after a diploid-type internal duplication event. From analyzing the partial duplication patterns, several evolutionary pathways were recognized for 6-tms TM proteins, i.e. from primordial 2-tms, 3-tms and 4-tms TM proteins to extant 6-tms proteins. Similarly, the duplication pattern analysis revealed plausible evolution scenarios that 7-tms TM proteins have arisen from 3-tms, 4-tms and 5-tms TM protein precursors via partial internal gene duplications. PMID- 15123418 TI - Polarity effects in the lactose operon of Escherichia coli. AB - An intergenic RNA segment between lacY and lacA of the lactose operon in Escherichia coli is cleaved by RNase P, an endoribonuclease. The cleavage of the intergenic RNA was ten times less efficient than cleavage of a tRNA precursor in vitro. Fragments of the RNase P cleavage product are detectable in vivo in the wild-type strain but not in a mutant strain at the restrictive temperature. The cleavage product that contains lacA in the wild-type strain was quickly degraded. When this intergenic segment was cloned upstream of a reporter gene, the expression of the reporter gene was also inhibited substantially in wild-type E.coli, but not in a temperature sensitive mutant strain in RNase P at the restrictive temperature. These results support data regarding the natural polarity between lacZ versus lacA, the downstream gene. PMID- 15123419 TI - Architecture and folding mechanism of the Azoarcus Group I Pre-tRNA. AB - Self-splicing RNAs must evolve to function in their specific exon context. The conformation of a group I pre-tRNA(ile) from the bacterium Azoarcus was probed by ribonuclease T(1) and hydroxyl radical cleavage, and by native gel electrophoresis. Biochemical data and three-dimensional models of the pre-tRNA showed that the tRNA is folded, and that the tRNA and intron sequences form separate tertiary domains. Models of the active site before steps 1 and 2 of the splicing reaction predict that exchange of the external G-cofactor and the 3' terminal G is accomplished by a slight conformational change in P9.0 of the Azoarcus group I intron. Kinetic assays showed that the pre-tRNA folds in minutes, much more slowly than the intron alone. The dependence of the folding kinetics on Mg(2+) and the concentration of urea, and RNase T(1) experiments showed that formation of native pre-tRNA is delayed by misfolding of P3-P9, including mispairing between residues in P9 and the tRNA. Thus, although the intron and tRNA sequences form separate domains in the native pre-tRNA, their folding is coupled via metastable non-native base-pairs. This could help prevent premature processing of the 5' and 3' ends of unspliced pre-tRNA. PMID- 15123420 TI - Dynamics of DNA loop capture by the SfiI restriction endonuclease on supercoiled and relaxed DNA. AB - The SfiI endonuclease is a prototype for DNA looping. It binds two copies of its recognition sequence and, if Mg(2+) is present, cuts both concertedly. Looping was examined here on supercoiled and relaxed forms of a 5.5 kb plasmid with three SfiI sites: sites 1 and 2 were separated by 0.4 kb, and sites 2 and 3 by 2.0 kb. SfiI converted this plasmid directly to the products cut at all three sites, though DNA species cleaved at one or two sites were formed transiently during a burst phase. The burst revealed three sets of doubly cut products, corresponding to the three possible pairings of sites. The equilibrium distribution between the different loops was evaluated from the burst phases of reactions initiated by adding MgCl(2) to SfiI bound to the plasmid. The short loop was favored over the longer loops, particularly on supercoiled DNA. The relative rates for loop capture were assessed after adding SfiI to solutions containing the plasmid and MgCl(2). On both supercoiled and relaxed DNA, the rate of loop capture across 0.4 kb was only marginally faster than over 2.0 kb or 2.4 kb. The relative strengths and rates of looping were compared to computer simulations of conformational fluctuations in DNA. The simulations concurred broadly with the experimental data, though they predicted that increasing site separations should cause a shallower decline in the equilibrium constants than was observed but a slightly steeper decline in the rates for loop capture. Possible reasons for these discrepancies are discussed. PMID- 15123421 TI - Bubble nucleation and cooperativity in DNA melting. AB - Bubbles in DNA are related to fundamental processes such as duplication and transcription. Using a new ensemble technique to trap intermediate states, we present direct measurements of the average length of the denaturation bubble and the statistical weights of the bubble states in the temperature-driven melting of DNA oligomers. For a bubble flanked by double-stranded regions, we find a nucleation size of approximately 20 bases, and a broad distribution of bubble sizes. However, for bubbles opening at the ends of the molecule there is no nucleation threshold. The measured statistical weights of different conformations agree with the predictions of the thermodynamic models in the case of unzipping from the ends; however, internal bubble states are not completely described by the models. The measurements further show that, due to end effects, the melting transition becomes a two-state process only in the limit of a molecule length L approximately 1 bp. PMID- 15123422 TI - A description scheme of biological processes based on elementary bricks of action. AB - With the fast growth of high-throughput strategies in Biology, there is a strong need to accelerate knowledge acquisition and organization of molecular functions. Unfortunately, although we know that there is a correlation between protein molecules and their functions, we are unable to clearly identify this link. Here, we revisit the current views of protein functions as well as their annotation, and we show that they are incompatible with unambiguous interpretations and the use of this knowledge. We describe herein a description scheme for biological processes based on elementary bricks of action that may be associated with biological molecules. To retrieve the descriptive quality found in annotations of other kinds of biological data, it was decided to develop a scheme involving four levels of abstraction: Basic Elements of Action, Biological Activities, Biological Functionalities and Biological Roles. This multi-level organization is a generic method; it allows for a description of biological processes by using a limited number of elementary bricks of action. Moreover, by using this description of biological processes, it should now be possible to clearly identify unambiguous relationships between the organization of biological processes and the structural or functional organizations of biological molecules. PMID- 15123423 TI - Loop relaxation, a mechanism that explains the reduced specificity of rabbit 20alpha-hydroxysteroid dehydrogenase, a member of the aldo-keto reductase superfamily. AB - The aldo-keto reductase rabbit 20alpha-hydroxysteroid dehydrogenase (rb20alpha HSD; AKR1C5) is less selective than other HSDs, since it exerts its activity both on androgens (C19 steroids) and progestins (C21 steroids). In order to identify the molecular determinants responsible for this reduced selectivity, binary (NADPH) and ternary (NADP(+)/testosterone) complex structures were solved to 1.32A and 2.08A resolution, respectively. Inspection of the cofactor-binding cavity led to the identification of a new interaction between side-chains of residues His222 and Lys270, which cover the central phosphate chain of the cofactor, reminiscent of the "safety-belt" found in other aldo-keto reductases. Testosterone is stabilized by a phenol/benzene tunnel composed of side-chains of numerous residues, among which Phe54, which forces the steroid to take up an orientation markedly contrasting with that found in HSD ternary complexes reported. Combining structural, site-directed mutagenesis, kinetic and fluorescence titration studies, we found that the selectivity of rb20alpha-HSD is mediated by (i) the relaxation of loop B (residues 223-230), partly controlled by the nature of residue 230, (ii) the nature of the residue found at position 54, and (iii) the residues found in the C-terminal tail of the protein especially the side-chain of the amino acid 306. PMID- 15123424 TI - Structural studies of the Nudix hydrolase DR1025 from Deinococcus radiodurans and its ligand complexes. AB - We have determined the crystal structure, at 1.4A, of the Nudix hydrolase DR1025 from the extremely radiation resistant bacterium Deinococcus radiodurans. The protein forms an intertwined homodimer by exchanging N-terminal segments between chains. We have identified additional conserved elements of the Nudix fold, including the metal-binding motif, a kinked beta-strand characterized by a proline two positions upstream of the Nudix consensus sequence, and participation of the N-terminal extension in the formation of the substrate-binding pocket. Crystal structures were also solved of DR1025 crystallized in the presence of magnesium and either a GTP analog or Ap(4)A (both at 1.6A resolution). In the Ap(4)A co-crystal, the electron density indicated that the product of asymmetric hydrolysis, ATP, was bound to the enzyme. The GTP analog bound structure showed that GTP was bound almost identically as ATP. Neither nucleoside triphosphate was further cleaved. PMID- 15123425 TI - A Structural-informatics approach for tracing beta-sheets: building pseudo C(alpha) traces for beta-strands in intermediate-resolution density maps. AB - We report the development of two computational methods to assist density map interpretation at intermediate resolutions: sheettracer for building pseudo C(alpha) models of beta-sheets, and a deconvolution method for enhancing features attributed to major secondary structural elements. Sheettracer is tightly coupled with sheetminer, which was developed to locate sheet densities in intermediate resolution density maps. The results from sheetminer are used as inputs to sheettracer, which employs a multi-step ad hoc morphological analysis of sheet densities to trace individual strands of beta-sheets. The methods were tested on simulated density maps from 12 protein crystal structures that represent a reasonably complete sampling of sheet morphology. The sheet-tracing results were quantitatively assessed in terms of sensitivity, specificity and rms deviations. Furthermore, sheettracer and the deconvolution method were rigorously tested on experimental maps of the lambda2 protein of reovirus at resolutions of 7.6A and 11.8A. Our results clearly demonstrate the capability of sheettracer in building pseudo-C(alpha) models of beta-sheets in intermediate-resolution density maps and the power of the deconvolution method in enhancing the performance of sheettracer. These computational methods, along with other related ones, should facilitate recognition and analysis of folding motifs from experimental data at intermediate resolutions. PMID- 15123427 TI - Unique physicochemical profile of beta-amyloid peptide variant Abeta1-40E22G protofibrils: conceivable neuropathogen in arctic mutant carriers. AB - A new early-onset form of Alzheimer's disease (AD) was described recently where a point mutation was discovered in codon 693 of the beta-amyloid (Abeta) precursor protein gene, the Arctic mutation. The mutation translates into a single amino acid substitution, glutamic acid-->glycine, in position 22 of the Abeta peptide. The mutation carriers have lower plasma levels of Abeta than normal, while in vitro studies show that Abeta1-40E22G protofibril formation is significantly enhanced. We have explored the nature of the Abeta1-40E22G peptide in more detail, in particular the protofibrils. Using size-exclusion chromatography (SEC) and circular dichroism spectroscopy (CD) kinetic and secondary structural characteristics were compared with other Abeta1-40 peptides and the Abeta12-28 fragment, all having single amino acid substitutions in position 22. We have found that Abeta1-40E22G protofibrils are a group of comparatively stabile beta sheet-containing oligomers with a heterogeneous size distribution, ranging from >100 kDa to >3000 kDa. Small Abeta1-40E22G protofibrils are generated about 400 times faster than large ones. Salt promotes their formation, which significantly exceeds all the other peptides studied here, including the Dutch mutation Abeta1 40E22Q. Position 22 substitutions had significant effects on aggregation kinetics of Abeta1-40 and in Abeta12-28, although the qualitative aspects of the effects differed between the native peptide and the fragment, as no protofibrils were formed by the fragments. The rank order of protofibril formation of Abeta1-40 and its variants was the same as the rank order of the length of the nucleation/lag phase of the Abeta12-28 fragments, E22V>E22A?E22G>E22Q?E22, and correlated with the degree of hydrophobicity of the position 22 substituent. The molecular mass of peptide monomers and protofibrils were estimated better in SEC studies using linear rather than globular calibration standards. The characteristics of the Abeta1-40E22G suggest an important role for the peptide in the neuropathogenesis in the Arctic form of AD. PMID- 15123426 TI - Regulation of LIP level and ROS formation through interaction of H-ferritin with G-CSF receptor. AB - A variety of hematopoietic factors including granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF), interleukin 3 (IL-3) and thrombopoietin (TPO) induce a rapid increase of intracellular reactive oxygen species (ROS). ROS induces the activation of many signaling molecules, including Shc, Lck, syk, PKC, MAPK, STAT3, through inhibition of protein phosphatase. Each growth factor has a specific cell-surface receptor, which activates both unique and shared signal transduction pathways. The processes of signal transduction linking cell-surface receptor to the formation of intracellular ROS have not been elucidated fully. Ferritins are composed of two subunit types, H and L, and made of 24 subunits that sequester up to 4500 atoms of iron. When the stored iron atoms are released from H-ferritin, through iron-catalyzed reaction, they have the capacity to promote the formation of ROS. Here, the interaction of G-CSFR and H-ferritin was confirmed by yeast two hybrid screen, mammalian two-hybrid assays, glutathione-S-transferase (GST) pull down experiments and immunoprecipitation studies in vitro and in vivo. Additional immunofluorescence assay showed that the two proteins colocalized along the plasma membrane and partly in the cytoplasm. The binding site for H-ferritin was demonstrated to locate to the box3 motif on the C-terminal region of granulocyte colony-stimulating factor receptor (G-CSFR). Furthermore, we found the interaction of full-length G-CSFR with H-ferritin was dissociated at 30 minutes after G-CSF induction and then began to assemble at 45 minutes. The labile iron pool (LIP) is a pool of redox-active iron complexes, which is regulated tightly by the expression of H-ferritin. Experiments showed that the level of LIP increased significantly at 30 minutes after G-CSF stimulation and intracellular ROS formation changed in a pattern similar to LIP response to G-CSF in bone marrow hematopoietic cells. G-CSF-induced changes in the level of LIP and ROS formation could be blocked by pretreatment with iron chelators that repressed the expression of H-ferritin. In addition, the phosphorylation of STAT3 induced by G CSF was decreased in iron chelator-treated hematopoietic cells. These data suggested that LIP may be released from the dissociated H-ferritin, and then induce intracellular ROS formation in the bone-marrow hematopoietic cells. ROS, acting as a second messenger, might take part in G-CSF receptor signal transduction. So, here, a new G-CSFR-H-ferritin-LIP-ROS pathway is proposed for regulation of intracellular ROS formation in bone-marrow hematopoietic cells. PMID- 15123428 TI - Crystal structure of the oxidized form of the periplasmic mercury-binding protein MerP from Ralstonia metallidurans CH34. AB - In Ralstonia metallidurans CH34, the gene merP encodes for a periplasmic mercury binding protein which is capable of binding one mercury atom. The metal-binding site of MerP consists of the highly conserved sequence GMTCXXC found in the family that includes metallochaperones and metal-transporting ATPases. We purified MerP from R.metallidurans CH34 and solved its crystal structure under the oxidized form at 2.0A resolution. Superposition with structures of other metal-binding proteins shows that the global structure of R.metallidurans CH34 oxidized MerP follows the general topology of the whole family. The largest differences are observed with the NMR structure of oxidized Shigella flexneri MerP. Detailed analysis of the metal-binding site suggests a direct role for Y66 in stabilizing the thiolate group of C17 during the mercury-binding reaction. The metal-binding site of oxidized MerP is also similar to the metal-binding sites of oxidized copper chaperone for superoxide dismutase and Atx1, two copper-binding proteins from Saccharomyces cerevisiae. Finally, the packing of the MerP crystals suggests that F38, a well-conserved residue in the MerP family may be important in mercury binding and transfer. We propose a possible mechanism of mercury transfer between two CXXC motifs based on a transient bi-coordinated mercury intermediate. PMID- 15123429 TI - Structural analysis of the human Golgi-associated plant pathogenesis related protein GAPR-1 implicates dimerization as a regulatory mechanism. AB - The plant pathogenesis related proteins group 1 (PR-1) and a variety of related mammalian proteins constitute a PR-1 protein family that share sequence and structural similarities. GAPR-1 is a unique family member as thus far it is the only PR-1 family member that is not co-translationally targeted to the lumen of the endoplasmic reticulum before trafficking to either vacuoles or secretion. Here we report that GAPR-1 may form dimers in vitro and in vivo, as determined by yeast two-hybrid screening, biochemical and biophysical assays. The 1.55A crystal structure demonstrates that GAPR-1 is structurally homologous to the other PR-1 family members previously solved (p14a and Ves V 5). Through an examination of inter-molecular interactions between GAPR-1 molecules in the crystal lattice, we propose a number of the highly conserved amino acid residues of the PR-1 family to be involved in the regulation of dimer formation of GAPR-1 with potential implications for other PR-1 family members. We show that mutagenesis of these conserved amino acid residues leads to a greatly increased dimer population. A recent report suggests that PR-1 family members may exhibit serine protease activity and further examination of the dimer interface of GAPR-1 indicates that a catalytic triad similar to that of serine proteases may be formed across the dimer interface by residues from both molecules within the dimer. PMID- 15123430 TI - Conformational properties of the iso-1-cytochrome C denatured state: dependence on guanidine hydrochloride concentration. AB - Production of seven single surface histidine variants of yeast iso-1-cytochrome c allowed measurement of the apparent pK(a), pK(a)(obs), for histidine-heme loop formation for loops of nine to 83 amino acid residues under varying denaturing conditions (2 M to 6 M guanidine hydrochloride, gdnHCl). A linear correlation between pK(a)(obs) and the log of the loop size is expected for a random coil, pK(a)(obs) proportional to k log(n), where k is a scaling factor and n is the number of monomers in the loop. For small loops of nine, 16, and 22 monomers, no dependence of pK(a)(obs) on loop size was observed at any denaturant concentration indicating effects from chain stiffness. For larger loops of 37, 56, 72, and 83 monomers, the dependence of pK(a)(obs) on log(n) was linear and the slope of that dependence decreased with increasing concentration of denaturant. The scaling factor obtained at 5 M and 6 M gdnHCl for the larger loop sizes was approximately -2.0, close to the value of -2.2 expected for a random coil with excluded volume. However, scaling factors obtained under less harsh denaturing conditions (2 M to 4.5 M gdnHCl) deviated strongly from that expected for a random coil, being in the range -3 to -4. The gdnHCl dependence of pK(a)(obs) at each loop size was also evaluated to obtain denaturant m-values. Short loops where chain stiffness dominates had similar m-values of approximately 0.25 kcal/mol M. For larger loops m-values decrease with increasing loop size indicating that less hydrophobic area is sequestered when larger loops form. It is known that the earliest events in protein folding involve the formation of simple loops. The data from these studies provide direct insight into the relative probability with which loops of different sizes will form, as well as the factors which affect loop formation. PMID- 15123431 TI - Phi value analysis of an allosteric transition of GroEL based on a single-pathway model. AB - There are currently two contradictory models for the kinetics of the ATP-induced GroEL allosteric transition occurring around 20 microM ATP. One model, proposed by Horovitz et al. demonstrates the existence of two parallel pathways for the allosteric transition and an abrupt ATP-dependent switch from one pathway to the other. The other model, which was proposed by the present authors, shows no need to assume the parallel pathways, and a combination of the transition-state theory and the Monod-Wyman-Changeux model of allostery can explain the kinetics as well as the equilibrium of the transition. The discrepancy appears to be due to whether we regard the transition as reversible or irreversible. Thus, here we have investigated the reversibility of the allosteric transition between 0 microM and 70 microM ATP by the use of a stopped-flow double-jump technique, which has allowed us to monitor the kinetics of the reverse reaction from the relaxed state at a high ATP concentration to the tense state at a low ATP concentration. The tryptophan fluorescence of a tryptophan-inserted variant of GroEL was used to follow the kinetics. As a result, the allosteric transition was shown to be a reversible process, supporting the validity of our model. We also show that the structural environment around the ATP-binding site of GroEL in the transition state is very similar to that in the relaxed state (Phi=0.9) by using a Phi value analysis in the kinetic Monod-Wyman-Changeux model, which is analogous to the mutational Phi value analysis in protein folding. PMID- 15123432 TI - Unique structural characteristics of peptide deformylase from pathogenic bacterium Leptospira interrogans. AB - Peptide deformylase (PDF), which is essential for normal growth of bacteria but not for higher organisms, is explored as an attractive target for developing novel antibiotics. Here, we present the crystal structure of Leptospira interrogans PDF (LiPDF) at 2.2A resolution. To our knowledge, this is the first crystal structure of PDF associating in a stable dimer. The key loop (named the CD-loop: amino acid residues 66-76) near the active-site pocket adopts "closed" or "open" conformations in the two monomers forming the dimer. In the closed subunit, the CD-loop and residue Arg109 block the entry of the substrate-binding pocket, while the active-site pocket of the open subunit is occupied by the C terminal tail from the neighbouring molecule. Moreover, a formate group, as one product of deformylisation, is observed bound with the active-site zinc ion. LiPDF displays significant structural differences in the C-terminal region compared to both type-I and type-II PDFs, suggesting a new family of PDFs. PMID- 15123434 TI - Predicting functional sites in proteins: site-specific evolutionary models and their application to neurotransmitter transporters. AB - Currently there exist several computational methods for predicting the functional sites in a set of homologous proteins based on their sequences. Due to difficulties in defining the functional site in a protein, it is not trivial to compare the performance of these methods, evaluate their limitations and quantify improvements by new approaches. Here, we use extensive mutation data from two proteins, Lac repressor and subtilisin, to perform such an analysis. Along with the evaluation of existing approaches, we describe a site class model of evolution as a tool to predict functional sites in proteins. The results indicate that this model, which simulates the evolution process at the amino acid level using site-specific substitution matrices, provides the most accurate information on functional sites in a given protein family. Secondly, we present an application of this model to neurotransmitter transporters, a superfamily of proteins of which we have limited experimental knowledge. Based on this application we present testable hypotheses regarding the mechanism of action of these proteins. PMID- 15123433 TI - The oxalate effect on release of iron from human serum transferrin explained. AB - A unique feature of the mechanism of iron binding to the transferrin (TF) family is the synergistic relationship between metal binding and anion binding. Little or no iron will bind to the protein without concomitant binding of an anion, physiologically identified as carbonate. Substitution of oxalate for carbonate produces no significant changes in polypeptide folding or domain orientation in the N-lobe of human serum TF (hTF) as revealed by our 1.2A structure. The oxalate is able to bind to the iron in a symmetric bidentate fashion, which, combined with the low pK(a) of the oxalate anion, makes iron displacement more difficult as documented by both iron release kinetic and equilibrium data. Characterization of an N-lobe in which the arginine at position 124 is mutated to alanine reveals that the stabilizing effect of oxalate is even greater in this mutant and nearly cancels the destabilizing effect of the mutation. Importantly, incorporation of oxalate as the synergistic anion appears to completely inhibit removal of iron from recombinant full-length hTF by HeLa S(3) cells, strongly indicating that oxalate also replaces carbonate in the C-lobe to form a stable complex. Kinetic studies confirm this claim. The combination of structural and functional data provides a coherent delineation of the effect of oxalate binding on hTF and rationalizes the results of many previous studies. In the context of iron uptake by cells, substitution of carbonate by oxalate effectively locks the iron into each lobe of hTF, thereby interfering with normal iron metabolism. PMID- 15123435 TI - Structural adaptation of the glycophorin A transmembrane homodimer to D-amino acid modifications. AB - Protein-protein recognition is an essential process in life. The chemistry of these kind of interactions is predominantly stereospecific (i.e. receptor-ligand, antibody-hapten binding). Here, we investigated whether the hydrophobic nature of the membrane affects this stereospecificity. To this end, we synthesized a diastereomer analogue (2D-GPA) of the glycophorin A transmembrane helix, with two l-valine residues replaced by their d-enantiomer. This ensures a disruption of the secondary structure. We investigated the ability of the diastereomer peptide to recognize the GPA chimera in the ToxR homodimer reporting system, in vivo. The peptide demonstrated a dose-dependent dominant negative effect on the GPA transmembrane in the bacterial ToxR system, suggesting a wild-type like interaction. This result was corroborated in vitro by fluorescence energy transfer between 2D-GPA and all-l GPA. Peptide binding to the bacteria was confirmed through confocal imaging, and Western blot confirmed that ToxR GPA receptor levels are not affected by the presence of the exogenous peptide. In order to understand the structural basis for heterodimer formation, homodimer and heterodimer structures, based on the NMR 3D structure of GPA, were subjected to a molecular dynamics simulation. The resulting heterodimer structure maintained most of the original inter-helical interactions, and its structure is similar to that of the homodimer. We postulate that the need to satisfy all H-bonds can compensate for the structural strain induced by the presence of the d-amino acid residues. PMID- 15123436 TI - Dietary fat consumption, readiness to change, and ethnocultural association in midlife African American women. AB - Cardiovascular disease (CVD) is the number 1 killer of American women, with African American women disproportionately represented. Dietary fat consumption is a major risk factor for the development of CVD. This study examined the average daily diet of urban midlife African American women, specifically the relation between dietary fat, readiness to change, ethnocultural association, and selected sociodemographic variables. Three hundred days of dietary data were collected. Sixty five percent of the participants reported currently avoiding high fat food with another 25% planning to avoid high fat food. Although 90% of the participants were avoiding or planning to avoid high fat foods, 77% were consuming diets with over 30% of their calories from fat. Of 11 variables considered, ethnocultural association was the only variable found to be consistently positively related to dietary fat intake. Community dietary education for midlife African American women needs to target all, especially those with stronger cultural bonds. PMID- 15123437 TI - Kansas office-based nurses' evaluation of patient tobacco cessation activities. AB - In this study, we examined the tobacco cessation efforts of nurses working in primary care settings. A 43-item questionnaire was mailed to 1,036 office-based nurses located throughout Kansas. With a response rate of 50.1%, 415 questionnaires were available for analysis. Although 89% of respondents encountered patients who smoked on a daily or weekly basis, only 51% reported documenting their patients' tobacco use, and 38% assessed patients' readiness to quit. Two thirds (66%) of nurses believed that tobacco management was part of their role but only 35% provided cessation advice, 23% recommended nicotine replacement therapy, and 14% provided coping techniques. Nurses cited barriers such as perceiving patients as disinterested or unmotivated to quit (65%) and having little time (55%), skills (32%), or knowledge (25%). Most (91%) agreed that they needed additional tobacco control education. Nurses who were advanced registered nurse practitioners or clinical nurse specialists were more likely to feel confident about their smoking cessation counseling skills compared to nurses with less education (66.7 vs. 31.2%, p =.010). Office-based nurses identified specific barriers that could be addressed through professional education about tobacco management. PMID- 15123438 TI - Applying the social ecological theory to type 2 diabetes prevention and management. AB - Obesity and Type 2 diabetes have become major public health problems in the United States. Community health nurses, with expertise in preventive health care, have the potential to play a vital role in addressing these significant health issues. The purposes of this article are to identify current challenges related to obesity and Type 2 diabetes and to present the social ecological theory as a framework for the expansion of the reach of diabetes prevention and management that is relevant to community health nurses. PMID- 15123439 TI - Public health nurses make a difference: a Japanese rural village benefits. PMID- 15123440 TI - HIV medication adherence programs: the importance of social support. AB - Since the advent of medical treatments for HIV, the promotion of adherence to these difficult treatment regimens has proven critical to disease management. Three Connecticut state-funded HIV medication adherence programs were evaluated.1 The purpose of this process evaluation was to explore and compare the goals and modality of each adherence program, assess client and staff satisfaction, and provide recommendations for the improvement of these programs. Focus group interviews with clients and individual interviews with staff were conducted at each of the programs. Interviews were transcribed, coded and analyzed with a code and retrieve method of theme identification. Focus group themes included the importance of social support on medication adherence and the "lifesaving" effect the program has had. The staff expressed that although complete adherence should be the long-term objective, more intermediate objectives should be considered (e.g., behavioral changes to increase clients' ability, self-esteem, and self efficacy to take medications). PMID- 15123443 TI - Online analysis of volatile organic compound emissions from Sitka spruce (Picea sitchensis). AB - Volatile organic compound (VOC) emissions from Sitka spruce (Picea sitchensis Bong.) growing in a range of controlled light and temperature regimes were monitored online with a proton transfer reaction-mass spectrometer (PTR-MS) operating at a temporal resolution of approximately 1 min. Isoprene emissions accounted for an average of more than 70% of measured VOCs and up to 3.5% of assimilated carbon. Emission rates (E) for isoprene correlated closely with photosynthetic photon flux (PPF) and temperature, showing saturation at a PPF of between 300 and 400 micromol m(-2) s(-1) and a maximum between 35 and 38 degrees C. Under standard conditions of 30 degrees C and 1000 micromol m(-2) s(-1) PPF, the mean isoprene E was 13 microg gdm(-1) h(-1), considerably higher than previously observed in this species. Mean E for acetaldehyde, methanol and monoterpenes at 30 degrees C were 0.37, 0.78 and 2.97 microg gdm(-1) h(-1), respectively. In response to a sudden light to dark transition, isoprene E decreased exponentially by > 98% over about 3 h; however, during the first 7 min, this otherwise steady decay was temporarily but immediately depressed to approximately 40% of the pre-darkness rate, before rallying during the following 7 min to rejoin the general downward trajectory of the exponential decay. The sudden sharp fall in isoprene E was mirrored by a burst in acetaldehyde E. The acetaldehyde E maximum coincided with the isoprene E minimum (7 min post illumination), and ceased when isoprene emissions resumed their exponential decay. The causes of, and linkages between, these phenomena were investigated. PMID- 15123444 TI - Transgenic poplar characterized by ectopic expression of a pine cytosolic glutamine synthetase gene exhibits enhanced tolerance to water stress. AB - Physiological responses to water stress in hybrid poplar (INRA 7171-B4, Populus tremula L. x P. alba L.) lines transformed to overexpress a pine cytosolic glutamine synthetase (GS1) gene were compared with those of non-transgenic plants. Before, during and after a drought treatment, net photosynthetic rates (Anet) were higher in transgenic than in non-transgenic plants. Stomatal conductance (gs) was higher in transgenic than in non-transgenic plants before, but not after exposure to drought. Before drought treatment, a sudden reduction in photosynthetic photon flux caused a greater burst of CO2 efflux in transgenic than non-transgenic plants, indicating greater photorespiratory activity. Drought caused greater reductions in photochemical quenching, photosystem II (PSII) antennae transfer efficiency (Fv'/Fm') and light-adapted PSII yield (PhiPSII) in non-transgenic than in transgenic plants, especially at low irradiances. Antennae based thermal dissipation was higher in transgenic plants than in non-transgenic plants both during the imposition of drought and 1 or 3 days after the relief of drought. Under severe water stress and subsequently, transgenic plants maintained a higher expression of glutamine synthetase, glutamate synthase and Rubisco and higher concentrations of chlorophyll and glycine than non-transgenic plants. These findings indicate that overexpression of pine cytosolic GS1 enhanced sustained photosynthetic electron transport capacity during severe stomatal limitation. The data also suggest that ectopic expression of cytosolic GS increases photorespiratory activity, and that this serves as a protective sink for electrons from photosynthetic reaction centers. PMID- 15123445 TI - Spatial and seasonal variability of temperature responses of biochemical photosynthesis parameters and leaf nitrogen content within a Pinus densiflora crown. AB - We measured seasonal variation in area-based nitrogen concentration (N), maximum rate of carboxylation (Vcmax) and maximum rate of electron transport (Jmax) in 1 year-old needles along four first-order branches within a Pinus densiflora Sieb. & Zucc. crown, and analyzed their relationships to growth irradiance and temperature. Each leaf light environment was expressed as a ratio of the monthly mean of daily integrated photosynthetically active irradiance (Iint) for the particular needle to Iint above the canopy (Irel). Needle N decreased in the upper crown during the development of new needles, whereas it remained fairly constant in the lower crown, reflecting differences between upper and lower crown needles in their contribution to the nitrogen of new needles. Gradients of N within the crown were correlated with Irel in all seasons (r2 = 0.40-0.78). Seasonal variation in N was weakly correlated with mean daily air minimum temperatures. Both Vcmax and Jmax showed seasonal variation in all first-order branches, and decreased to their lowest values in winter. The gradients of Vcmax and Jmax within the crown were not correlated with Irel in some seasons, but were correlated with changes in N in most months (r2 = 0.33-0.75), except in the winter. Furthermore, the regression slope of the relationship between N and Vcmax and the temperature response of Vcmax and Jmax exhibited seasonal variation. PMID- 15123446 TI - Size dependency of photosynthetic water- and nitrogen-use efficiency and hydraulic limitation in Acer mono. AB - We examined open-grown Acer mono Maxim. trees of different sizes to test the hypotheses that (1) hydraulic limitation increases with tree size, thereby reducing photosynthesis, and (2) photosynthetic water- and nitrogen-use efficiencies change with tree size. Maximum net assimilation rate per unit dry mass was significantly lower in large trees than in small trees, whereas leaf nitrogen concentration increased with tree size. As a consequence, photosynthetic nitrogen-use efficiency decreased with increase in tree size. Photosynthetic water-use efficiency, however, increased with tree size, partly as a result of reduced stomatal conductance. Neither root-to-leaf hydraulic conductance nor minimum leaf water potential changed with tree size. PMID- 15123447 TI - Age-related changes in ecosystem structure and function and effects on water and carbon exchange in ponderosa pine. AB - As forests age, their structure and productivity change, yet in some cases, annual rates of water loss remain unchanged. To identify mechanisms that might explain such observations, and to determine if widely different age classes of forests differ functionally, we examined young (Y, approximately 25 years), mature (M, approximately 90 years) and old (O, approximately 250 years) ponderosa pine (Pinus ponderosa Dougl. ex P. Laws.) stands growing in a drought-prone region of central Oregon. Although the stands differed in tree leaf area index (LAIT) (Y = 0.9, M = 2.8, O = 2.1), cumulative tree transpiration measured by sap flow did not differ substantially during the growing season (100-112 mm). Yet when water was readily available, transpiration per unit leaf area of the youngest trees was about three times that of M trees and five times that of O trees. These patterns resulted from a nearly sixfold difference in leaf specific conductance (KL) between the youngest and oldest trees. At the time of maximum transpiration in the Y stand in May-June, gross carbon uptake (gross ecosystem production, GEP) was similar for Y and O stands despite an almost twofold difference in stand leaf area index (LAIS). However, the higher rate of water use by Y trees was not sustainable in the drought-prone environment, and between spring and late summer, KL of Y trees declined fivefold compared with a nearly twofold decline for M trees and a < 30% reduction in O trees. Because the Y stand contained a significant shrub understory and more exposed soil, there was no appreciable difference in mean daily latent energy fluxes between the Y stand and the older stands as measured by the eddy-covariance technique. These patterns resulted in 60 to 85% higher seasonal GEP and 55 to 65% higher water-use efficiency at the M and O stands compared with the Y stand. PMID- 15123448 TI - Long-term effects of fire frequency on carbon storage and productivity of boreal forests: a modeling study. AB - Climate change is predicted to shorten the fire interval in boreal forests. Many studies have recorded positive effects of fire on forest growth over a few decades, but few have modeled the long-term effects of the loss of carbon and nitrogen to the atmosphere. We used a process-based, dynamic, forest ecosystem model, which couples the carbon, nitrogen and water cycles, to simulate the effects of fire frequency on coniferous forests in the climate of Prince Albert, Saskatchewan. The model was calibrated to simulate observed forest properties. The model predicted rapid short-term recovery of net primary productivity (NPP) after fire, but in the long term, supported the hypotheses that (1) current NPP and carbon content of boreal forests are lower than they would be without periodic fire, and (2) any increase in fire frequency in the future will tend to lower NPP and carbon storage. Lower long-term NPP and carbon storage were attributable to (1) loss of carbon on combustion, equal to about 20% of NPP over a 100-200 year fire cycle, (2) loss of nitrogen by volatilization in fire, equal to about 3-4 kg N ha(-1) year(-1) over a 100-200 year fire cycle, and (3) the fact that the normal fire cycle is much shorter than the time taken for the forest (especially the soil) to reach an equilibrium carbon and nitrogen content. It was estimated that a shift in fire frequency from 200 to 100 years over 1000 Mha of boreal forest would release an average of about 0.1 Gt C year(-1) over many centuries. PMID- 15123449 TI - Physiological, morphological and allocational plasticity in understory deciduous trees: importance of plant size and light availability. AB - In a 4-year study, we investigated changes in leaf physiology, crown morphology and whole-tree biomass allocation in seedlings and saplings of shade-tolerant sugar maple (Acer saccharum Marsh.) and intermediate shade-tolerant yellow birch (Betula alleghaniensis Britt.) growing in natural understory light (0.5 to 35% of full sunlight) or in understory light reduced by 50% with shade nets to simulate the effect of gap closure. Leaf physiological parameters were mainly influenced by the light gradient, whereas crown morphological and whole-tree allocational parameters were mainly influenced by tree size. No single physiological, morphological or allocational trait was identified that could explain the difference in shade tolerance between the species. Yellow birch had higher growth rates, biomass allocation to branches and leaf physiological plasticity and lower crown morphological plasticity in unmodified understory light than sugar maple. Sugar maple did not display significant physiological plasticity, but showed variation with tree size in both crown morphology and whole-tree biomass allocation. When sugar maple was small, a greater proportion of whole-tree biomass was allocated to roots. However, physiological differences between the species decreased with decreasing light and most morphological and allocational differences tended to disappear with increasing tree size, suggesting that many species differences in shade-tolerance are expressed mainly during the seedling stage. Understory trees of both species survived for 4 years under shade nets, possibly because of higher plasticity when small and the use of stored reserves when taller. PMID- 15123450 TI - Temperature effects on xylem sap osmolarity in walnut trees: evidence for a vitalistic model of winter embolism repair. AB - We studied the effect of temperature on the carbohydrate status of parenchyma cells during winter in relation to the efflux and influx of sugars between parenchyma cells and xylem vessels in 1-year-old twigs of walnut (Juglans regia L.). The mechanism of sugar transfer between contact cells and vessels was also investigated. We obtained new insights into the possible osmotic role of sugars, particularly sucrose, in stem pressure formation and winter embolism repair. Accumulation of sucrose in the xylem sap during winter was mainly influenced by: (1) abundant conversion of starch to sucrose in the symplast at low temperatures; (2) sucrose efflux into the apoplast at low temperatures (1 degrees C); and (3) inefficient sugar uptake at low temperatures, although efficient sugar uptake occurred at 15 degrees C. We hypothesize that a diethyl pyrocarbonate (DEPC) sensitive protein mediates facilitated diffusion of sucrose from parenchyma cells to xylem vessels (efflux) in walnut. We discuss the possible occurrence of active H+-sucrose symports and the coexistence of both influx and efflux processes in walnut in winter and the modulation of the relative importance of these flows by temperature. PMID- 15123451 TI - Characteristics of photosynthesis and stomatal conductance in the shrubland species manuka (Leptospermum scoparium) and kanuka (Kunzea ericoides) for the estimation of annual canopy carbon uptake. AB - Responses of photosynthesis to carbon dioxide (CO2) partial pressure and irradiance were measured on leaves of 39-year-old trees of manuka (Leptospermum scoparium J. R. Forst. & G. Forst.) and kanuka (Kunzea ericoides var. ericoides (A. Rich.) J. Thompson) at a field site, and on leaves of young trees grown at three nitrogen supply rates in a nursery, to determine values for parameters in a model to estimate annual net carbon uptake. These secondary successional species belong to the same family and commonly co-occur. Mean (+/- standard error) values of the maximum rate of carboxylation (hemi-surface area basis) (Vcmax) and the maximum rate of electron transport (Jmax) at the field site were 47.3 +/- 1.9 micromol m(-2) s(-1) and 94.2 +/- 3.7 micromol m(-2) s(-1), respectively, with no significant differences between species. Both Vcmax and Jmax were positively related to leaf nitrogen concentration on a unit leaf area basis, and the slopes of these relationships did not differ significantly between species or between the trees in the field and young trees grown in the nursery. Mean values of Jmax/Vcmax measured at 20 degrees C were significantly lower (P < 0.01) for trees in the field (2.00 +/- 0.05) than for young trees in the nursery with similar leaf nitrogen concentrations (2.32 +/- 0.08). Stomatal conductance decreased sharply with increasing air saturation deficit, but the sensitivity of the response did not differ between species. These data were used to derive parameters for a coupled photosynthesis-stomatal conductance model to scale estimates of photosynthesis from leaves to the canopy, incorporating leaf respiration at night, site energy and water balances, to estimate net canopy carbon uptake. Over the course of a year, 76% of incident irradiance (400-700 nm) was absorbed by the canopy, annual net photosynthesis per unit ground area was 164.5 mol m(-2) (equivalent to 1.97 kg C m(-2)) and respiration loss from leaves at night was 37.5 mol m(-2) (equivalent to 0.45 kg m(-2)), or 23% of net carbon uptake. When modeled annual net carbon uptake for the trees was combined with annual respiration from the soil surface, estimated net primary productivity for the ecosystem (0.30 kg C m(-2)) was reasonably close to the annual estimate obtained from independent mensurational and biomass measurements made at the site (0.22 +/- 0.03 kg C m(-2)). The mean annual value for light-use efficiency calculated from the ratio of net carbon uptake (net photosynthesis minus respiration of leaves at night) and absorbed irradiance was 13.0 mmol C mol(-1) (equivalent to 0.72 kg C GJ(-1)). This is low compared with values reported for other temperate forests, but is consistent with limitations to photosynthesis in the canopy attributable mainly to low nitrogen availability and associated low leaf area index. PMID- 15123452 TI - Stomatal conductance of Acer rubrum ecotypes under varying soil and atmospheric water conditions: predicting stomatal responses with an abscisic acid-based model. AB - A multiplicative model of stomatal conductance was developed and tested in two functionally distinct ecotypes of Acer rubrum L. (red maple). The model overcomes the main limitation of the commonly used Ball-Berry model (Ball et al. 1987) by accounting for stomatal behavior under soil drying conditions. We combined the Ball-Berry model with an integrated expression of abscisic acid (ABA)-based stomatal response to ABA concentration ([ABA]) in bulk leaf tissue (gfac), which coupled physiological changes at the leaf level with those in the root. The factor gfac = exp(-beta[ABA]L) incorporated the stomatal response to [ABA] into the Ball-Berry model by down regulating stomatal conductance (gs) in response to physiological changes in the root. The down regulation of gs is pertinent under conditions where soil drying may modify the delivery of chemical signals to leaf stomata. Model testing indicated that the multiplicative model was capable of predicting gs in red maple under wide ranges of soil and atmospheric conditions. Concordance correlation coefficients were high (between 0.59 and 0.94) for the tested ecotypes under three environmental conditions (atmospheric, rhizospheric and minimal stress). The study supported the use of gfac as a gas exchange function that controls water stress effects on gs and aids in the prediction of gs responses. PMID- 15123453 TI - Stomatal limitation to CO2 assimilation and down-regulation of photosynthesis in Quercus ilex resprouts in response to slowly imposed drought. AB - Holm oak (Quercus ilex L.) is native to hot, dry Mediterranean forests where limited water availability often reduces photosynthesis in many species, and forest fires are frequent. Holm oaks resprout after a disturbance, with improved photosynthetic activity and water relations compared with unburned plants. To better understand the role of water availability in this improvement, watering was withheld from container-grown plants, either intact (controls) or resprouts after excision of the shoot, to gradually obtain a wide range of soil water availabilities. At high water availability, gas exchange rates did not differ between controls and resprouts. At moderate soil dryness, net photosynthesis of control plants decreased as a result of increased stomatal limitation, whereas gas exchange rates of resprouts, which had higher midday and predawn leaf water potentials, were unchanged. Under severe drought, resprouts showed a less marked decline in gas exchange than controls and maintained photosystem II integrity, as indicated by chlorophyll fluorescence measurements. Photosynthesis was down regulated in both plant types in response to reduced CO2 availability caused by high stomatal limitation. Lower non-stomatal limitations in resprouts than in control plants, as evidenced by higher carboxylation velocity and the capacity for ribulose-1,5-bisphosphate regeneration, conferred greater drought resistance under external constraints similar to summer conditions at midday. PMID- 15123454 TI - Mist, substrate water potential and cutting water potential influence rooting of stem cuttings of loblolly pine. AB - We investigated the influence of cutting water potential (Psicut) on rooting of juvenile hardwood (dormant) and softwood (succulent) stem cuttings of loblolly pine (Pinus taeda L.) propagated under varying substrate water potentials (Psisub) and volumes of mist application. Mist treatment and Psisub contributed to the Psicut of unrooted stem cuttings. When Psisub was held constant across mist treatments, mist treatment contributed strongly to Psicut. Substrate water potential affected rooting percentage when mist treatment was sub-optimal or excessive, otherwise mist treatment had a stronger effect than Psisub on rooting percentage. Cuttings rooted best when subjected to moderate cutting water potentials (-0.5 to -1.2 MPa) during the initial 4 or 5 weeks of the rooting period. Cuttings experiencing either severe water deficit or no water deficit rooted poorly. We conclude that the rooting environment should impose a moderate water stress on loblolly pine stem cuttings to achieve optimum rooting. PMID- 15123455 TI - Effects of elevated concentrations of ozone and carbon dioxide on the electrical impedance of leaves of silver birch (Betula pendula) clones. AB - Effects of elevated concentrations of tropospheric ozone ([O3]) and carbon dioxide ([CO2]) on leaves of two silver birch (Betula pendula Roth) clones were monitored for three growing seasons (1998, 1999, 2000) by means of electrical impedance spectroscopy (EIS). The field trial with open-top chambers (OTCs) was conducted on two clones (Clone 4 and Clone 80) with five treatments and four independent replicates. Treatments were: (1) outside control, (2) chamber control, (3) 2x ambient [O3], (4) 2x ambient [CO2] and (5) 2x ambient [CO2] + 2x ambient [O3]. Fumigations started in 1999 and continued in 2000. Measurements were made in 1998 before the fumigations and thereafter EIS was carried out four times in each season. The impedance spectra of about 10 leaves from each tree at each time were measured at 42 frequencies between 80 and 1 MHz. Leaf spectra were modeled by a distributed circuit element model (DCE) (one DCE in series with a resistor), which yields the extracellular and intracellular resistances, the relaxation time and the distribution coefficient of the relaxation time. The EIS properties of the leaves changed significantly during the growing season when new leaves were expanding. The clones differed in their EIS properties. Clone 4 had a significantly higher extracellular resistance and distribution coefficient than Clone 80. The clones responded similarly to the fumigation treatments. Differences between treatments emerged especially during the second fumigation season in 2000. Elevated [O3] reduced both the relaxation time and the extracellular resistance, indicating cell membrane damage. Elevated [CO2] increased the intracellular resistance, indicating changes in symplastic composition. The biological interpretation of the EIS parameters in birch leaves is discussed. PMID- 15123456 TI - Belowground carbon dynamics in loblolly pine (Pinus taeda) immediately following diammonium phosphate fertilization. AB - Forest soils store an immense quantity of labile carbon (C) and a may be a large potential sink for atmospheric C. Forest management practices such as fertilization may enhance overall C storage in soils, yet changes in physiological processes following nutrient amendments have not been widely investigated. We intensively monitored belowground C dynamics for nearly 200 days following diammonium phosphate fertilization of pot-grown loblolly pine (Pinus taeda L.) seedlings in an effort to examine the short-term effects of fertilization on processes involved in soil C sequestration. Soil respiration rates initially increased in fertilized pots relative to controls, followed by a brief reversal in this trend and then a final sustained pattern of elevated rates of soil respiration in the fertilized treatment. Patterns in soil respiration rates over time reflected changes in autotrophic (root) and heterotrophic (microbial) components of soil respiration. Root respiration rates were greater in the fertilized treatment 49 days following fertilization and returned to control rates by the end of the study. In contrast, microbial respiration rates and microbial activity per soil C concentration remained depressed over the same time period. Compared with control seedlings, total root biomass was 27% greater in fertilized seedlings harvested at the end of the study, indicating that the elevated soil respiration rates observed toward the end of the study were a result of increased respiring root biomass. We conclude that fertilization, at least over the short-term, may increase soil C sequestration by increasing belowground biomass production and reducing microbial driven C turnover. PMID- 15123457 TI - Nondestructive detection of decay in living trees. AB - We used a four-point resistivity method to detect wood decay in living trees. A low-frequency alternating current was applied to the stem and the induced voltage measured between two points along the stem. The effective resistivity of the stem was estimated based on stem cross-sectional area. A comparison within a group of trees showed that trees with butt rot had an effective resistivity that was at least a factor of two lower than that of healthy trees. In tests on several groups of Norway spruce (Picea abies (L.) Karst.) comprising more than 300 trees in total, the method detected butt rot with high accuracy. We validated the method both by measurements and by finite element modeling and simulations. PMID- 15123458 TI - Sensitivity to chilling temperatures and distribution differ in the mangrove species Kandelia candel and Avicennia marina. AB - We compared the effects of short-term (hours) and long-term (days) exposure to chilling temperatures on the photosynthetic gas exchange, leaf characteristics and chlorophyll a fluorescence of seedlings of the mangrove species Kandelia candel Druce and Avicennia marina (Forsk.) Vierh. Both species occur along the west coast of Taiwan, but K. candel occurs further north than A. marina. We hypothesized that temperature was one of the major environmental factors limiting the northern distribution of A. marina. Avicennia marina was more sensitive to chilling temperatures than K. candel. Leaves of both species showed reductions in light-saturated photosynthetic rates (Amax), stomatal conductance (gs) and quantum yield of photosystem II after a 1-h exposure to 15 degrees C, with A. marina showing significantly greater reductions in Amax and gs than K. candel. No significant differences in Amax, gs and electron transport rate (ETR) were found between leaves of K. candel grown at 15 and 30 degrees C for 10 days. However, leaves of A. marina grown for 10 days at 15 degrees C had significantly lower Amax, gs and ETR than plants grown at 30 degrees C. After 20 days at 15 degrees C, leaf mass per area of both species was increased significantly, whereas area based chlorophyll concentrations were reduced, with significantly greater changes in A. marina than in K. candel. We concluded that sensitivity to low winter temperatures is a primary limiting factor in the distribution of A. marina along the western coast of Taiwan. PMID- 15123459 TI - Surgery and imatinib in the management of GIST: emerging approaches to adjuvant and neoadjuvant therapy. AB - Gastrointestinal stromal tumor (GIST) is a neoplasm of the gastrointestinal tract, mesentery, or omentum that expresses the protein-tyrosine kinase KIT (CD117) and is the most common mesenchymal tumor arising at these sites. Surgical resection is the first-line intervention for operable GISTs, particularly localized primary tumors, and it was historically the only effective treatment. However, more than half of all GIST patients present with locally advanced, recurrent, or metastatic disease. The 5-year survival rate ranges from 50% to 65% after complete resection of a localized primary GIST and decreases to approximately 35% for patients with advanced disease who undergo complete surgical resection. A total of 40% to 90% of all GIST surgical patients subsequently have postoperative recurrence or metastasis. Imatinib is a potent, specific inhibitor of KIT that has demonstrated significant activity and tolerability in the treatment of malignant unresectable or metastatic GIST, inducing tumor shrinkage of 50% or more or stabilizing disease in most patients. A key strategy for prolonging the survival of patients with GIST is to improve the outcome of surgery. It is possible that the adjuvant and neoadjuvant use of imatinib (e.g., rendering initially inoperable tumors resectable) in the overall management approach to advanced GIST may contribute to surgeons' success in attaining this objective. PMID- 15123460 TI - Artificial neural network for prediction of lymph node metastases in gastric cancer: a phase II diagnostic study. AB - BACKGROUND: Extension of lymphadenectomy in gastric cancer is controversial, and preoperative diagnosis of lymph node metastases (LNM) is difficult. Therefore, knowledge-based systems such as the Maruyama computer program (MCP) are being developed. MCP shows good prognostic value for the compartments; however, for different lymph node groups (LNG) there are a large number of false positives. The aim of this study was to evaluate artificial neural networks (ANN) for predicting LNM in patients with gastric cancer and to compare the predictive power with that of MCP. METHODS: A total of 135 consecutive patients who underwent D2 gastrectomy were included. We applied a single-layer perceptron to the data of 4302 patients from the National Cancer Center, Tokyo, and compared the results with those from the MCP. RESULTS: Prediction of N(+) or N0 with ANN-1 (Borrmann classification, T category, and tumor size and location) had an accuracy of 79%. The predictive value for LNM in each of the LNG varied: ANN-1, 64% to 86%; MCP, 42% to 70%. We constructed another ANN by using the additional parameter of metastases in LNG 3 as an example of sentinel node. The accuracy of this ANN was 93%. CONCLUSIONS: Using an ANN, LNM in each LNG can be accurately predicted. Additional knowledge about one lymph node improves the results. PMID- 15123461 TI - Treatment of peritoneal carcinomatosis from colorectal cancer: impact of complete cytoreductive surgery and difficulties in conducting randomized trials. AB - BACKGROUND: Colorectal peritoneal carcinomatosis (PC) is a frequent and very lethal event. However, cure may be possible with maximal cytoreductive surgery associated with early postoperative intraperitoneal chemotherapy (EPIC). METHODS: Between 1996 and 2000, we conducted a two-center prospective randomized trial comparing EPIC plus systemic chemotherapy with systemic chemotherapy alone, both after complete cytoreductive surgery of colorectal PC. Only 35 patients could be included among the 90 who were theoretically required, mainly because of patient dissatisfaction with the inclusion criteria. For this reason, the trial was stopped prematurely. RESULTS: Analysis of these 35 patients showed that complete resection of PC resulted in a 2-year survival rate of 60%-far above the classic 10% survival rate among patients with colorectal PC treated with systemic chemotherapy and symptomatic surgery. In this small series, EPIC did not demonstrate any advantage for survival. CONCLUSIONS: This supports the use of complete cytoreductive surgery in selected patients and calls for a prospective randomized trial comparing adjuvant systemic chemotherapy with intraperitoneal chemohyperthermia after complete resection. PMID- 15123462 TI - Diagnostic laparoscopy for primary and secondary liver malignancies: impact of improved imaging and changed criteria for resection. AB - BACKGROUND: Diagnostic laparoscopy (DL) combined with laparoscopic ultrasonography (LUS) has previously shown positive results as a staging modality for liver malignancies. Recent improvements in noninvasive diagnostic imaging techniques such as multiphasic spiral computed tomography, together with the policy that bilobar disease or the number of lesions is no longer considered an absolute exclusion criterion for curative resection, could reduce the additional value of DL. This study retrospectively analyzed the efficacy of DL combined with LUS for liver malignancies to assess the effect of improved imaging and changed criteria for resection. METHODS: All patients with primary or metachronous secondary liver malignancy eligible for resection in 1997 to 2002 were included. RESULTS: DL combined with LUS was performed in 84 consecutive patients (56 men and 28 women; mean age, 59 years) with primary (n = 33) or secondary (n = 51) liver malignancies. DL showed unresectability in 13 patients (39%) with primary malignancy. Exploratory laparotomy showed that an additional 5 (25%) of the remaining 20 patients had unresectable disease. DL showed unresectability in 5 patients (12%) with colorectal liver metastasis (n = 43). At laparotomy, another 7 (18%) of the remaining 38 patients had unresectable disease. In five patients (13%) from the latter group, LUS could not be performed because of adhesions from previous surgery. CONCLUSIONS: DL combined with LUS is an adequate staging modality for primary liver malignancies. For colorectal liver metastasis, more liberal resection criteria, a high failure rate due to adhesions from previous surgery, and better preoperative imaging probably resulted in a lower efficacy. PMID- 15123463 TI - Squamous cell carcinoma related oncogene regulates angiogenesis through vascular endothelial growth factor-A. AB - BACKGROUND: Squamous cell carcinoma related oncogene expression (SCCRO) correlates with vascular endothelial growth factor-A expression. This data is validated in human lung tumors and provides a putative pathway for angiogenesis in a subset of squamous cell carcinomas. Squamous cell carcinoma related oncogene is a novel oncogene identified by positional cloning of a recurrent amplification at 3q26.3. It is over-expressed in 39.8% of lung, head and neck, cervical, and ovarian carcinomas. SCCRO imparts an aggressive phenotype to affected cancers, which may be related to increased angiogenesis due to SCCRO expression. Our previous work has demonstrated a link between SCCRO and vascular endothelial growth factor-A (VEGF-A) expression in vitro, suggesting a mechanism for SCCRO induced angiogenesis. The present study aims to confirm and validate this link between SCCRO and VEGF-A expression in an ex vivo human tumor cohort. METHODS: Fresh tissue was collected at Memorial Sloan-Kettering Cancer Center from 34 patients undergoing primary resection of lung squamous cell carcinomas. RNA was extracted from this tissue, reverse-transcribed, and real-time polymerase chain reaction (RT-PCR) was carried out using a BioRad iQ iCycler with SYBR green fluorophore. Microvessel counting was performed on the tumor specimens using CD34 immunohistochemistry. RESULTS: The expression of both SCCRO and VEGF-A mRNA varies widely in both tumor and normal tissue. SCCRO and VEGF-A co-expression was significantly correlated (R(2) = 0.63; P < 0.032). Microvessel counts were not associated with expression of SCCRO or VEGF-A and failed to significantly predict survival. VEGF-A expression in this patient group is a predictor of overall survival (P < 0.032). CONCLUSIONS: VEGF-A expression correlates with SCCRO expression in these primary human lung squamous cell carcinomas and is a predictor of clinical behavior. This data supports the association of SCRRO and VEGF-A in the induction of angiogenesis. PMID- 15123464 TI - A prospective analysis of the effect of blue-dye volume on sentinel lymph node mapping success and incidence of allergic reaction in patients with breast cancer. AB - BACKGROUND: This study examined whether the volume of isosulfan blue dye used in sentinel lymph node (SLN) mapping in breast cancer is related to the SLN identification rate or to the incidence of allergic reactions. METHODS: From January 2001 to November 2002, 1728 breast cancer patients underwent 1832 SLN mapping procedures with the combined technique of intraparenchymal blue dye and intradermal radioisotope. Details of each procedure and all allergic reactions were prospectively recorded. Bilateral synchronous SLN procedures were considered as one dye exposure but as two distinct procedures for determining mapping success. Dye-only success was defined as the proportion of cases in which the SLN was identified by blue dye alone. Overall dye success was defined as the proportion of cases in which the SLN was identified by blue dye with or without isotope. RESULTS: When stratified by volume of blue dye, there were no significant differences in dye-only successes, overall dye successes, or mapping failures. Allergic reactions were documented in 31 (1.8%) of 1728 patients. Hypotensive reactions occurred in 3 (.2%) of 1728 patients; 2 (.1%) required pressor support. There was a nonsignificant trend toward fewer allergic reactions with smaller volumes of blue dye. CONCLUSIONS: In combined-technique SLN mapping protocols for breast cancer, using smaller volumes of blue dye may represent a means of optimizing the safety of the procedure without compromising its success. PMID- 15123465 TI - Cryoablation of early-stage breast cancer: work-in-progress report of a multi institutional trial. AB - BACKGROUND: With recent improvements in breast imaging, our ability to identify small breast tumors has markedly improved, prompting significant interest in the use of ablation without surgical excision to treat early-stage breast cancer. We conducted a multi-institutional pilot safety study of cryoablation in the treatment of primary breast carcinomas. METHODS: Twenty-nine patients with ultrasound-visible primary invasive breast cancer 1.5 cm, cryoablation was not reliable with this technique. Patients with noncalcified DCIS were the cause of most cryoablation failures. CONCLUSIONS: Cryoablation is a safe and well-tolerated office-based procedure for the ablation of early-stage breast cancer. At this time, cryoablation should be limited to patients with invasive ductal carcinoma 75%) for the matched sets with a few exceptions, including questions that asked for a provider's own behavior or expectation (eg, expecting further contact with a patient's family). However, chance-corrected agreement (measured as kappa) was low. CONCLUSIONS: There was a good quality of care at the time of death for most patients, with minimal pain and suffering reported by providers. There were differences in perception explained by roles (physicians vs nurses). Future research should examine differences by provider experience and in other care settings. PMID- 15123475 TI - How pediatricians counsel parents when no "best-choice" management exists: lessons to be learned from hypoplastic left heart syndrome. AB - BACKGROUND: Hypoplastic left heart syndrome (HLHS) is a life-threatening congenital cardiac defect. Three mutually exclusive management options exist: the Norwood palliative procedure, cardiac transplantation, and comfort care without surgical intervention. OBJECTIVES: To assess which management options are presented to parents of infants with HLHS, and to determine what factors influence physicians' recommendations. DESIGN: Cross-sectional survey. SETTING: Fourteen of the largest pediatric cardiac surgery centers in the United States. PARTICIPANTS: Attending physicians in neonatology, cardiology, critical care practice, and cardiac surgery. INTERVENTION: A survey was distributed asking physicians what options they present to parents of infants with HLHS and what their recommendations are in general, as well as physician perceptions of HLHS outcomes and demographic information. MAIN OUTCOME MEASURES: Which options physicians discuss and which they recommend. RESULTS: Of 454 eligible physicians 257 (57%) responded to the survey, of which 110 make treatment recommendations to parents. Neonatologists were least likely to recommend surgery. Physicians who recommend surgery recommend procedures performed at their own institution over those performed elsewhere (odds ratio, 2.80; 95% confidence interval, 2.24-3.51). Twenty-six percent of physicians do not discuss nonsurgical management, and 25% of those at centers that do not perform cardiac transplantation do not discuss this option. The recommendations physicians make to parents are poorly associated with their predictions of postoperative outcomes. CONCLUSIONS: Physician recommendations to parents are poorly associated with their estimates of outcomes, and some physicians recommend the treatment preferred at their own institution over other options even when they predict better outcomes from another approach. Further, many physicians do not disclose all reasonable management options to parents of infants with HLHS. These findings raise doubts as to whether parents are given adequate information to make truly informed decisions. PMID- 15123476 TI - Trends from an HIV seroprevalence study among childbearing women in New York State from 1988 through 2000: a valuable epidemiologic tool. AB - BACKGROUND: Women in New York State are heavily affected by the human immunodeficiency virus (HIV) epidemic. New York has had the largest number of births to HIV-infected pregnant women in the United States. Data collected as part of the Survey of Childbearing Women have been valuable for assessing the impact of the disease on the women of New York. OBJECTIVE: To assess HIV prevalence trends among childbearing women in New York State. DESIGN, SETTING, AND PARTICIPANTS: An unlinked HIV seroprevalence study was conducted among all women residing in and giving birth in New York State from 1988 through 2000. Trend and cohort analyses were conducted. Main Outcome Measure HIV prevalence, defined as the number of HIV-positive specimens divided by the total number of HIV-positive and HIV-negative specimens, by geographic region, racial/ethnic group, and maternal age cohort. RESULTS: Trends indicated a steady decline in HIV prevalence in New York State. New York City had a 49% decrease in prevalence between 1988 through 1989 and 1999 through 2000, and the rest of the state showed a 24% decline. However, birth cohort analysis indicated different patterns in trend by subpopulation, with some groups experiencing little or no decline. CONCLUSION: This study reports on the only statewide population-based HIV prevalence data currently available for childbearing women; these data have been a valuable tool for monitoring trends, targeting resources, and evaluating programs and policies. PMID- 15123477 TI - Identifying children at high risk for overweight at school entry by weight gain during the first 2 years. AB - OBJECTIVE: To assess the best anthropometric predictor from birth to 2 years for later overweight, based on recent studies reporting that large infant weight or length gain predicts subsequent overweight. DESIGN: Retrospective cohort study. SETTING: Southern Germany. PARTICIPANTS: German children (n = 4235) aged 5.0 to 6.9 years. MAIN OUTCOME MEASURES: Overweight at school entry was defined according to sex- and age-specific body mass index cutpoints proposed by the International Obesity Task Force. Weight, length, body mass index, and ponderal index differences between birth, 6 months, 12 months, and 24 months of age were compared by receiver operating characteristic curves and predictive values. RESULTS: For all variables, the largest area under the receiver operating characteristic curve was observed with a 24-month follow-up: 0.76 (95% confidence interval [CI], 0.74-0.79) for weight, 0.70 (95% CI, 0.67-0.72) for body mass index, and 0.58 (95% CI, 0.55-0.61) for length gain. The highest Youden index ([sensitivity plus specificity] minus 1) for weight gain from birth to 24 months (41%) was attained for a cutpoint of 9764 g, with a corresponding positive likelihood ratio of 2.39 (95% CI, 2.20-2.59) and positive predictive value of 19% (95% CI, 17%-21%), despite an odds ratio of 5.7 (95% CI, 4.5-7.1). CONCLUSIONS: Weight gain from birth to 24 months was the best overall predictor of later overweight compared with other anthropometric markers and intervals. However, the corresponding poor positive predictive value suggests that only 1 of 5 children with a large weight gain in the first 2 years is overweight at school entry and reflects an insufficient predictability in the general population. PMID- 15123478 TI - Analysis of perpetrator admissions to inflicted traumatic brain injury in children. AB - BACKGROUND: Scientific and courtroom debate exists regarding the timing of onset of symptoms and the mechanism of injury in infants and children with inflicted traumatic brain injury (ITBI). OBJECTIVES: To determine the time interval between ITBI and the onset of symptoms and to explore the mechanism of ITBI. DESIGN, SETTING, AND PATIENTS: Retrospective review of all cases of pediatric ITBI admitted between January 1, 1981, and July 31, 2001, to a large academic medical center and cases admitted to 2 additional academic institutions between January 1, 1996, and August 31, 2000, and January 1, 2001, and July 31, 2001, comparing 81 cases of ITBI in which perpetrators admitted to abuse with 90 cases in which no abuse admission was made. The patients with perpetrator admissions to ITBI consisted of 53 boys (65%) and 28 girls (35%). Their ages ranged from 2 weeks to 52 months. MAIN OUTCOME MEASURES: Characteristics associated with perpetrator admissions to ITBI in children. RESULTS: Shaking was the most common mechanism of injury among all cases with perpetrator admissions: 55 (68%) of the 81 perpetrators admitted to shaking the children. Impact was not described in 44 (54%) of the 81 cases. In cases in which only impact was described, 60% (12/20) of the children showed skull or scalp injury, compared with 12% (4/32) with skull or scalp injury in the shake only group. In 52 (91%) of 57 cases in which the time to the onset of symptoms was described, symptoms appeared immediately after the abuse. In 5 cases (9%), the timing of symptoms was less clear, but they occurred within 24 hours. None of the children were described as behaving normally after the event. CONCLUSIONS: The symptoms of inflicted head injury in children are immediate. Most perpetrators admitted to shaking without impact. These data, combined with the relative lack of skull and scalp injury, suggest that shaking alone can produce the symptoms seen in children with ITBI. PMID- 15123479 TI - Benefits of a school-based asthma treatment program in the absence of secondhand smoke exposure: results of a randomized clinical trial. AB - BACKGROUND: Daily maintenance medications are recommended for all children with mild persistent to severe persistent asthma; however, poor adherence to these medications is common. OBJECTIVE: To evaluate the impact of school-based provision of inhaled corticosteroids on asthma severity among urban children with mild persistent to severe persistent asthma. DESIGN: Children aged 3 to 7 years with mild persistent to severe persistent asthma were identified at the start of the 2000-2001 and 2001-2002 school years in Rochester. Children were assigned randomly to a school-based care group (daily inhaled corticosteroids provided through the school) or a usual-care group (inhaled corticosteroids not given through school). MAIN OUTCOME MEASURE: Improvement in parent-reported symptom free days. RESULTS: Of 242 eligible children, 184 were enrolled from 54 urban schools. Data for 180 children were available. Parents of children in the school based care group had a greater improvement in quality of life compared with parents of children in the usual-care group (change score, 0.63 vs 0.24; P =.047); also, children in the school-based care group vs the usual-care group missed less school because of asthma (mean total days missed, 6.8 vs 8.8; P =.047) and experienced more symptom-free days during the early winter months (mean days per 2-week period, 9.2 vs 7.3; P =.02). A post hoc analysis revealed that all significant findings were produced by differences among children who were not exposed to secondhand smoke. Furthermore, among children not exposed to smoke, those in the school-based care group vs the usual-care group had more symptom-free days overall (11.5 vs 10.5; P =.046), had fewer days needing rescue medications (1.6 vs 2.3; P =.03), and were less likely to have had 3 or more acute visits for asthma (6 [13%] of 47 children vs 17 [31%] of 54 children; P =.03). CONCLUSIONS: School-based provision of inhaled corticosteroids significantly improved symptoms, quality of life, and absenteeism among urban children with mild persistent to severe persistent asthma. This effect was seen only among children not exposed to secondhand smoke. PMID- 15123480 TI - Interpreting subgroup analyses: is a school-based asthma treatment program's effect modified by secondhand smoke exposure? PMID- 15123481 TI - The association of caffeinated beverages with blood pressure in adolescents. AB - OBJECTIVE: To assess the association between the consumption of caffeinated beverages and blood pressure in African American and white adolescents. DESIGN: This study was part of ongoing research examining stress-induced hemodynamic responses in adolescents. African American and white adolescents (n = 159) selected foods and beverages for a 3-day sodium-controlled diet. Caffeine in these foods was used to stratify participants into 3 categories (0-50 mg/d, >50 100 mg/d, and >100 mg/d). Before menu selection, blood pressure readings were obtained. STATISTICAL ANALYSIS: A general linear model (multiple regression with both categorical and continuous variables) was developed to assess the effects of race, category of caffeine intake, and interaction of race and caffeine intake on systolic and diastolic blood pressure controlling for sex and body mass index (calculated as weight in kilograms divided by height in meters squared). RESULTS: The association between systolic blood pressure and caffeine category varied by race (P =.001). African Americans consuming more than 100 mg/d of caffeine had higher systolic blood pressure readings than the groups consuming 0 to 50 mg/d (mean difference, 6.0 mm Hg; 95% confidence interval [CI], 2.3 to 9.7) or more than 50 to 100 mg/d (mean difference, 7.1 mm Hg; 95% CI, 3.4 to 10.7). The effect on diastolic blood pressure was less pronounced (P =.08). The diastolic blood pressure of the group consuming more than 100 mg/d was 3.7 mm Hg (95% CI, 0.41 to 7.0) higher than the group consuming more than 50 to 100 mg/d and was not statistically different from the group consuming 0 to 50 mg/d (mean difference, 2.4 mm Hg; 95% CI, -0.9 to 5.8). There was no evidence that the association between diastolic blood pressure and caffeine intake varied by race (P =.80). CONCLUSIONS: For adolescents, especially African American adolescents, caffeine intake may increase blood pressure and thereby increase the risk of hypertension. Alternatively, caffeinated drink consumption may be a marker for dietary and lifestyle practices that together influence blood pressure. Additional research is needed owing to rising rates of adolescent hypertension and soft drink consumption. PMID- 15123482 TI - Bulimia nervosa in adolescents: a disorder in evolution? AB - BACKGROUND: There are few reports that describe the manifestation of bulimia nervosa (BN) among adolescents. Moreover, none make reference to the comparative clinical manifestation of adolescent BN and adolescent anorexia nervosa. Nor are any reports available of how distinct partial-syndrome BN cases are from those that meet full diagnostic criteria for BN. OBJECTIVES: To describe 3 groups of adolescents, those with a full-syndrome eating disorder (BN and anorexia nervosa) or partial-syndrome BN, and to compare these groups along demographic, general psychopathology, and eating disorder variables. DESIGN: The study population included 120 adolescents with eating disorders who were initially seen at The University of Chicago Eating Disorders Program, Chicago, Ill, for treatment. All participants completed an assessment prior to treatment. MEASURES: Weight and height were obtained from all participants. Participants also completed a baseline demographic questionnaire (eg, menstrual status, ethnicity, family status), Beck Depression Inventory, Rosenberg Self-Esteem Scale, and the Eating Disorder Examination. RESULTS: Partial-syndrome BN cases are clinically quite similar to their full-syndrome counterparts. Only objective binge eating episodes and purge frequency distinguished BN and partial-syndrome BN cases. Anorexia nervosa cases, on the other hand, were quite distinct from BN and partial syndrome BN cases on almost all variables. CONCLUSION: Early recognition and swift treatment of eating disorders in adolescents, regardless of whether a diagnostic threshold is met, are imperative because they will lead to early intervention thereby potentially improving eating disorder recovery rates. PMID- 15123483 TI - Enteral vs intravenous rehydration therapy for children with gastroenteritis: a meta-analysis of randomized controlled trials. AB - OBJECTIVE: To review the relative efficacy and safety of enteral vs intravenous (IV) rehydration therapy in treating childhood gastroenteritis. DATA SOURCES: MEDLINE, EMBASE, and the Cochrane Controlled Trials Register databases were searched. Known investigators and expert bodies were contacted to locate unpublished and ongoing studies. STUDY SELECTION: Studies were selected based on the following criteria: randomized or quasi-randomized trials; children younger than 15 years with a clinical diagnosis of gastroenteritis of less than 1-week duration; interventions comprising enteral and IV treatment arms; and at least 1 of the following: major adverse event rates, treatment failure rates, weight gain with treatment, measurement of ongoing losses, length of hospital stay, costs of treatment, and satisfaction with treatment. DATA EXTRACTION: Data were extracted from eligible studies, which were then combined using a random-effects model. DATA SYNTHESIS: Sixteen trials involving 1545 children and conducted in 11 countries were identified. Compared with children treated with IV rehydration, children treated with oral rehydration had significantly fewer major adverse events, including death or seizures (relative risk, 0.36; 95% confidence interval [CI], 0.14-0.89), and a significant reduction in length of hospital stay (mean, 21 hours; 95% CI, 8-35 hours). There was no difference in weight gain between the 2 groups (mean, -26 g; 95% CI, -61 to 10 g). The overall failure rate of enteral therapy was 4.0% (95% CI, 3.0%-5.0%). CONCLUSIONS: For childhood gastroenteritis, enteral rehydration is as effective if not better than IV rehydration. Enteral rehydration by the oral or nasogastric route is associated with significantly fewer major adverse events and a shorter hospital stay compared with IV therapy and is successful in most children. PMID- 15123484 TI - Osteopenia in children who have undergone posterior fossa or craniospinal irradiation for brain tumors. AB - OBJECTIVES: To determine lumbar spine and total body bone mineral density (BMD) in pediatric patients who have undergone cranial or craniospinal irradiation for posterior fossa tumors, specifically medulloblastoma and ependymoma and to analyze the association between degree of osteopenia and factors that may affect BMD. METHODS: Retrospective and prospective data collection included medical record review and examination, including pubertal, dietary, and activity assessment. Lumbar spine and total body BMD were measured by means of dual energy x-ray absorptiometry. Patients were routinely observed by the endocrinology department, and hormone deficiencies were corrected promptly. A subset of patients received calcium and vitamin D supplementation and underwent repeat BMD measurement 1 year later. RESULTS: Of 24 patients aged 4 to 20 years, 11 of whom were male, recruited from 1996 through 1999, 19 had medulloblastoma. All 19 underwent craniospinal radiotherapy plus a boost to the posterior fossa (mean +/- SD of 5410 +/- 130 rad [54.1 +/- 1.3 Gy] to the posterior fossa, mean +/- SD of 3470 +/- 460 rad [34.7 +/- 4.6 Gy] to the whole brain and spinal axis), and 8 of 19 underwent chemotherapy. The remaining 5 patients had ependymoma and underwent irradiation to the posterior fossa only (mean +/- SD of 5680 +/- 720 rad [56.8 +/ 7.2 Gy]). Therefore, there were 3 treatment groups: craniospinal irradiation and chemotherapy, only craniospinal irradiation, and only posterior fossa irradiation. Bone mineral studies were performed a mean +/- SD of 5.42 +/- 3.23 years after therapy. Our patients had lower total body BMD (mean z score, -0.47; 95% confidence interval, -0.85 to -0.09) and lumbar spine BMD (mean z score, 1.27; 95% confidence interval, -1.81 to -0.73) as compared with those of the the general population. There was no significant difference in mean lumbar spine BMD between patients in the 3 groups. Our patients consumed a diet deficient in vitamin D and calcium (mean +/- SD 53.6% +/- 24.1% and 70.0% +/- 37.4% of the amount recommended, respectively). Of 7 patients who underwent measurements 1 year later, 5 had in increase in BMD that was parallel to normal curves, with no compensatory increase. Four patients were hypothyroid, 6 were growth hormone deficient, and 6 were both. All hormones were replaced, with the exception of growth hormone in 1 patient. By using regression analysis, the factors that affected lumbar spine BMD, protectively in both cases, were calcium intake (beta = 0.015, 95% confidence interval, 0.001-0.029) and female sex (beta = 1.422, 95% confidence interval, 0.456-2.388). CONCLUSIONS: Children who have undergone irradiation for posterior fossa tumors have diminished total body and lumbar spine BMD, as compared with those of the general population. This reduction was similar within all 3 treatment groups, which suggests that chemotherapy did not play a major role and that localized irradiation may have systemic effects. This population often has balance and gait problems, so the risk of falling, coupled with osteopenia, may place them at considerably increased risk of fractures. PMID- 15123485 TI - Depiction of tobacco use in popular children's picture books. PMID- 15123486 TI - Prevalence of narcotic analgesic abuse among students: individual or polydrug abuse? PMID- 15123487 TI - The nightmare. PMID- 15123488 TI - The effect of a managed behavioral health carve-out on quality of care for medicaid patients diagnosed as having schizophrenia. AB - CONTEXT: Managed behavioral health carve-outs (MBHCOs) are a regular feature of public and private mental health care systems and have been successful in reducing costs. The evidence on quality impacts is limited and suggests comparable quality overall, except that people with severe psychiatric disorders may be those most disadvantaged by MBHCOs. OBJECTIVE: To explore the effect of implementing an MBHCO on the quality of outpatient care received by enrollees diagnosed as having schizophrenia. DESIGN AND PARTICIPANTS: Observational retrospective cohort study using a quasi-experimental design of state Medicaid enrollees diagnosed as having schizophrenia, aged 18 to 64 years between 1994 and 2000 in the carve-out and comparison regions (8082 person-years). SETTING: Ambulatory care. MAIN OUTCOME MEASURES: Quality indicators derived from the Schizophrenia Patient Outcomes Research Team recommendations. RESULTS: There was no statistical difference between the carve-out and integrated arrangements in the likelihood of receiving any antipsychotic medication (odds ratio [OR], 1.02; 95% confidence interval [CI], 0.81-1.29), second-generation antipsychotics (including clozapine: OR, 1.05; 95% CI, 0.86-1.28; not including clozapine: OR, 1.05; 95% CI, 0.85-1.29), or antiextrapyramidal medication (OR, 1.36; 95% CI, 0.84-2.19). The carve-out was negatively associated with receiving any individual therapy (OR, 0.27; 95% CI, 0.22-0.33), group therapy (OR, 0.19; 95% CI, 0.14 0.25), and psychosocial rehabilitation (OR, 0.31; 95% CI, 0.26-0.38). Family therapy occurred for less than 1% of this population in both carve-out and integrated regions. CONCLUSIONS: The MBHCO was not associated with changes in medication quality (for which it was not at financial risk). It was significantly associated with sharp decreases in the likelihood of receiving psychosocial treatments (for which it was financially at risk)-independent of whether a clinical evidence base supported them. PMID- 15123489 TI - Brain metabolic alterations in medication-free patients with bipolar disorder. AB - BACKGROUND: Bipolar disorder (BD) has substantial morbidity and incompletely understood neurobiological underpinnings. OBJECTIVE: To investigate brain chemistry in medication-free individuals with BD. DESIGN: Two-dimensional proton echo-planar spectroscopic imaging (PEPSI) (32 x 32, 1-cm(3) voxel matrix) acquired axially through the cingulate gyrus was used to quantify regional brain chemistry. SETTING: The Center for Anxiety and Depression at the University of Washington in Seattle and the Bipolar Research Programs at McLean Hospital and the Massachusetts General Hospital in Boston. PARTICIPANTS: Thirty-two medication free outpatients with a diagnosis of BD type I (BDI) or BD type II (BDII), predominantly in a depressed or mixed-mood state, were compared with 26 age- and sex-matched healthy controls. MAIN OUTCOME MEASURES: Tissue type (white and gray) and regional analyses were performed to evaluate distribution of lactate; glutamate, glutamine, and gamma-aminobutyric acid (Glx); creatine and phosphocreatine (Cre); choline-containing compounds (Cho); N-acetyl aspartate; and myo-inositol. Chemical relationships for diagnosis and mood state were evaluated. RESULTS: Patients with BD exhibited elevated gray matter lactate (P =.005) and Glx (P =.007) levels; other gray and white matter chemical measures were not significantly different between diagnostic groups. Isolated regional chemical alterations were found. An inverse correlation between 17-item Hamilton Depression Rating Scale scores and white matter Cre levels was observed for BD patients. CONCLUSIONS: Gray matter lactate and Glx elevations in medication-free BD patients suggest a shift in energy redox state from oxidative phosphorylation toward glycolysis. The possibility of mitochondrial alterations underlying these findings is discussed and may provide a theoretical framework for future targeted treatment interventions. PMID- 15123490 TI - Four-year prospective outcome and natural history of mania in children with a prepubertal and early adolescent bipolar disorder phenotype. AB - BACKGROUND: Diagnosis of child mania has been contentious. OBJECTIVE: To investigate natural history and prospective validation of the existence and long episode duration of mania in children. DESIGN: Four-year prospective longitudinal study of 86 subjects with intake episode mania who were all assessed at 6, 12, 18, 24, 36, and 48 months. The phenotype was defined as DSM-IV bipolar I disorder (manic or mixed) with at least 1 cardinal symptom (elation and/or grandiosity) to ensure differentiation from attention-deficit/hyperactivity disorder. Parent and child informants were separately interviewed, by highly experienced research nurses, using the Washington University in St Louis Kiddie Schedule for Affective Disorders and Schizophrenia (WASH-U-KSADS). A Children's Global Assessment Scale score of 60 or less was needed to establish definite impairment. Treatment was by subjects' community practitioners. SETTING: Research unit in a university medical school. PARTICIPANTS: Subjects were obtained from psychiatric and pediatric sites by consecutive new case ascertainment, and their baseline age was 10.8 +/- 2.7 years. Onset of the baseline episode was 7.4 +/- 3.5 years. (Data are given as mean +/- SD.) MAIN OUTCOME MEASURES: Episode duration, weeks ill, recovery/relapse rates, and outcome predictors. RESULTS: Prospective episode duration of manic diagnoses, using onset of mania as baseline date, was 79.2 +/- 66.7 consecutive weeks. Any bipolar disorder diagnosis occurred during 67.1% +/- 28.5% of total weeks, during the 209.4 +/- 3.3 weeks of follow-up. Subjects spent 56.9% +/- 28.8% of total weeks with mania or hypomania (unipolar or mixed), and 38.7% +/- 28.8% of these were with mania. Major or minor depression and dysthymia (unipolar or mixed) occurred during 47.1% +/- 30.4% of total weeks. Polarity switches occurred 1.1 +/- 0.7 times per year. Low maternal warmth predicted faster relapse after recovery from mania (chi(2) = 13.6, P =.0002), and psychosis predicted more weeks ill with mania or hypomania (F(1,80) = 12.2, P =.0008). Pubertal status and sex were not predictive. (Data are given as mean +/- SD.) CONCLUSIONS: These findings validate the existence, long-episode duration, and chronicity of child mania. Differences from the natural history of adult bipolar disorder are discussed. PMID- 15123491 TI - Familiality of symptom dimensions in depression. AB - BACKGROUND: Depression is a clinically heterogeneous disorder thought to result from multiple genes interacting with environmental and developmental components. A dimensional rather than a categorical approach to depressive phenotype definition may be more useful for identification of susceptibility genes. OBJECTIVES: To perform an exploratory factor analysis on a range of depressive and anxiety symptoms in a large, well-defined sample of depressed siblings, as well as a confirmatory factor analysis in a separate large group of unrelated depressed subjects, and to analyze correlations of identified symptom dimensions between depressed siblings. DESIGN: Subjects (N = 1034), including 475 sibling pairs, with a history of at least 2 depressive episodes were recruited from the Depression Network Study, a large-scale multicenter collection of families affected by recurrent unipolar depression. Subjects were interviewed using the Schedules for Clinical Assessment in Neuropsychiatry (SCAN) and diagnosed according to the DSM-IV and the International Classification of Diseases, 10th Revision, using a computerized scoring program (CATEGO5). Factor analysis was carried out on 26 depression symptom items, including 4 anxiety screening items. Confirmatory factor analysis was performed on an independent sample of 485 depressed individuals. RESULTS: Four interpretable factors were identified: (1) mood symptoms and psychomotor retardation; (2) anxiety; (3) psychomotor agitation, guilt, and suicidality; and (4) appetite gain and hypersomnia. For each symptom group, a quantitative scale was constructed, and correlations between siblings were calculated. There was a moderate degree of sibling homotypia for some depressive symptoms, and factors 1, 2, and 3 showed significant positive familial correlation (0.145 [P =.001], 0.335 [P<.001], and 0.362 [P<.001], respectively). CONCLUSIONS: This is the first study of large, well-defined samples of depressed subjects in whom symptom dimensions have been derived and then confirmed using independent material. The significant correlations between siblings for 3 of the dimensions suggest substantial familial, perhaps genetic, etiologies. PMID- 15123492 TI - Quality of care for medicaid-covered youth treated with antidepressant therapy. AB - BACKGROUND: Although antidepressant use has increased in pediatric populations, few studies have addressed the quality of follow-up care or duration of treatment for depressed youth. OBJECTIVE: To evaluate the quality of care for antidepressant-treated youth, using the Health Plan Employer Data and Information Set guidelines (>or=3 visits in the 3 months after a new antidepressant prescription fill and continuation of antidepressant use at 3 and 6 months) as a benchmark. DESIGN: Administrative records were examined for 1205 Medicaid-covered youth (aged 5-18 years) who presented with a "new episode" of depression in 1998. Statistics were generated to describe the number of follow-up visits and duration of treatment within 6 months of first prescription fill. RESULTS: A total of 507 (42.1%) youth with new episodes of depression were treated with antidepressants. Selective serotonin reuptake inhibitors accounted for 80.9% of prescriptions. Twenty-eight percent (28.1%) of youth with an antidepressant fill had 3 or more follow-up visits in the subsequent 3 months; however, an additional 29.2% had no further provider visits. Selective serotonin reuptake inhibitors were continued by 46.6% of treated youth at 3 months and by 26.3% at 6 months. CONCLUSIONS: Many antidepressant-treated youth do not receive adequate follow-up or duration of treatment. Future studies should address reasons for poor follow-up and methods to improve monitoring for these youth. PMID- 15123493 TI - Stress burden and the lifetime incidence of psychiatric disorder in young adults: racial and ethnic contrasts. AB - BACKGROUND: With the exception of studies of individual traumatic events, the significance of stress exposure in psychiatric disorder previously has not been effectively examined. OBJECTIVE: To address the hypothesis that accumulated adversity represents an important risk factor for the subsequent onset of depressive and anxiety disorders. DESIGN: A community-based study of psychiatric and substance use disorders among a large, ethnically diverse cohort representative of young adults in South Florida. Adversity was estimated with a count of major and potentially traumatic events experienced during one's lifetime and prior to the onset of disorder. SETTING: Most interviews took place in the homes of participants, with 30% conducted by telephone. PARTICIPANTS: We obtained a random sample of individuals aged 18 to 23 years from a previously studied representative sample of young adolescents. Because participants in the prior study were predominantly boys, a supplementary sample of girls was randomly obtained from the early-adolescence school class rosters. A total of 1803 interviews were completed, representing a success rate of 70.1%. MAIN OUTCOME MEASURES: Depressive and anxiety disorders were assessed through computer assisted personal interviews using the DSM-IV version of the Michigan Composite International Diagnostic Interview. RESULTS: Level of lifetime exposure to adversity was found to be associated with an increased risk of subsequent onset of depressive and/or anxiety disorder. This association remained clearly observable when childhood conduct disorder, attention-deficit/hyperactivity disorder, prior substance dependence, and posttraumatic stress disorder were held constant and when the possibility of state dependence effects was considered. CONCLUSION: Evidence suggests that high levels of lifetime exposure to adversity are causally implicated in the onset of depressive and anxiety disorders. PMID- 15123494 TI - Conduct problems in children and adolescents: a twin study. AB - BACKGROUND: Evidence supports a genetic influence on conduct problems as a continuous measure of behavior and as a diagnostic category. However, there is a lack of studies using a genetically informative design combined with several different informants and different settings. OBJECTIVES: To examine genetic and environmental influences on conduct problems rated by parent and teacher reports and self-reports and to determine whether their ratings reflect a common underlying phenotype. DESIGN: A twin study design was used to examine conduct problem scores from ratings by teachers, parents, and twins themselves. SETTING: General community. PARTICIPANTS: Twins aged 5 to 17 years participating in the Cardiff Study of All Wales and North England Twins (CaStANET) project. MAIN OUTCOME MEASURES: Conduct problem scale from the Strengths and Difficulties Questionnaire. RESULTS: Conduct problem scores were significantly heritable based on parent and teacher reports and self-reports. Combining data from all 3 informants showed that they are rating a common underlying phenotype of pervasive conduct problems that is entirely genetic, while teacher ratings show separate genetic influences that are not shared with other raters. CONCLUSIONS: Conduct problems are significantly heritable based on parent and teacher reports and self reports, and are also influenced by environmental effects that impinge uniquely on children from the same family. There is a cross-situational conduct problems' phenotype, underlying the behavior measured by all informants, that is wholly genetic in origin. No significant influence of shared environmental effects was found. PMID- 15123495 TI - A randomized effectiveness trial of stepped collaborative care for acutely injured trauma survivors. AB - CONTEXT: Although posttraumatic stress disorder (PTSD) and alcohol abuse frequently occur among acutely injured trauma survivors, few real-world interventions have targeted these disorders. OBJECTIVE: We tested the effectiveness of a multifaceted collaborative care (CC) intervention for PTSD and alcohol abuse. DESIGN: Randomized effectiveness trial. PARTICIPANTS: We recruited a population-based sample of 120 male and female injured surgical inpatients 18 or older at a level I trauma center. INTERVENTION: Patients were randomly assigned to the CC intervention (n = 59) or the usual care (UC) control condition (n = 61). The CC patients received stepped care that consisted of (1) continuous postinjury case management, (2) motivational interviews targeting alcohol abuse/dependence, and (3) evidence-based pharmacotherapy and/or cognitive behavioral therapy for patients with persistent PTSD at 3 months after injury. MAIN OUTCOME MEASURES: We used the PTSD symptomatic criteria (PTSD Checklist) at baseline and 1, 3, 6, and 12 months after injury, and alcohol abuse/dependence (Composite International Diagnostic Interview) at baseline and 6 and 12 months after injury. RESULTS: Random-coefficient regression analyses demonstrated that over time, CC patients were significantly less symptomatic compared with UC patients with regard to PTSD (P =.01) and alcohol abuse/dependence (P =.048). The CC group demonstrated no difference (-0.07%; 95% confidence interval [CI], -4.2% to 4.3%) in the adjusted rates of change in PTSD from baseline to 12 months, whereas the UC group had a 6% increase (95% CI, 3.1%-9.3%) during the year. The CC group showed on average a decrease in the rate of alcohol abuse/dependence of 24.2% (95% CI, -19.9% to -28.6%), whereas the UC group had on average a 12.9% increase (95% CI, 8.2%-17.7%) during the year. CONCLUSIONS: Early mental health care interventions can be feasibly and effectively delivered from trauma centers. Future investigations that refine routine acute care treatment procedures may improve the quality of mental health care for Americans injured in the wake of individual and mass trauma. PMID- 15123496 TI - Sleep in lifetime posttraumatic stress disorder: a community-based polysomnographic study. AB - BACKGROUND: Sleep complaints are common in posttraumatic stress disorder (PTSD) and are included in the DSM criteria. Polysomnographic studies conducted on small samples of subjects with specific traumas have yielded conflicting results. We therefore evaluated polysomnographic sleep disturbances in PTSD. METHODS: A representative cohort of young-adult community residents followed-up for 10 years for exposure to trauma and PTSD was used to select a subset for sleep studies for 2 consecutive nights and the intermediate day. Subjects were selected from a large health maintenance organization and are representative of the geographic area except for the extremes of the socioeconomic status range. The subset for the sleep study was selected from the 10-year follow-up of the cohort (n = 913 [91% of the initial sample]). Eligibility criteria included (1) subjects exposed to trauma during the preceding 5 years; (2) others who met PTSD criteria; and (3) a randomly preselected subsample. Of 439 eligible subjects, 292 (66.5%) participated, including 71 with lifetime PTSD. Main outcomes included standard polysomnographic measures of sleep induction, maintenance, staging, and fragmentation; standard measures of apnea/hypopnea and periodic leg movement; and results of the multiple sleep latency test. RESULTS: On standard measures of sleep disturbance, no differences were detected between subjects with PTSD and control subjects, regardless of history of trauma or major depression in the controls. Persons with PTSD had higher rates of brief arousals from rapid eye movement (REM) sleep. Shifts to lighter sleep and wake were specific to REM and were significantly different between REM and non-REM sleep (F(1,278) = 5.92; P =.02). CONCLUSIONS: We found no objective evidence for clinically relevant sleep disturbances in PTSD. An increased number of brief arousals from REM sleep was detected in subjects with PTSD. Sleep complaints in PTSD might represent amplified perceptions of brief arousals from REM sleep. PMID- 15123497 TI - Apolipoprotein E epsilon4 count affects age at onset of Alzheimer disease, but not lifetime susceptibility: The Cache County Study. AB - BACKGROUND: The incidence of Alzheimer disease (AD) increases strongly with age, but little is known about the cumulative incidence of AD over a lifetime of 100 years, or its relationship to the polymorphic APOE locus that encodes apolipoprotein E. APOE is a strong genetic risk factor for AD. OBJECTIVES: To estimate the occurrence of AD as a function of age and number of APOE epsilon4 alleles; and to explore evidence for heterogeneity of AD risk related to APOE genotype and to other sources. DESIGN: Nonparametric and parametric survival analyses of AD incidence in prospective longitudinal study. SETTING AND PARTICIPANTS: A total of 3308 elderly residents of Cache County, Utah. MAIN OUTCOME MEASURES: Cumulative incidence of AD; in mixture models assuming susceptible and nonsusceptible individuals, the proportion of individuals not susceptible to AD at any age. RESULTS: Models that assumed a proportion of invulnerable individuals provided strongly improved fit to the data. These models estimated the 100-year lifetime incidence of AD at 72%, implying that 28% of individuals would not develop AD over any reasonable life expectancy. We confirmed the acceleration of AD onset in individuals with 1 or, especially, 2 APOE, epsilon4 alleles but observed no meaningful difference in 100-year lifetime incidence related to number of epsilon4 alleles. CONCLUSIONS: The APOE epsilon4 allele acts as a potent risk factor for AD by accelerating onset. However, the risk of AD appears heterogeneous in ways independent of APOE. Some individuals seem destined to escape AD, even over an extended lifespan. Their relative invulnerability may reflect other genes or environmental factors that can be investigated. PMID- 15123498 TI - Depression: international intervention for a global problem. PMID- 15123499 TI - Cross-national differences in diet, the outcome of schizophrenia and the prevalence of depression: you are (associated with) what you eat. PMID- 15123500 TI - Foetal origins of schizophrenia: testable hypotheses of genetic and environmental influences. PMID- 15123501 TI - Global burden of depressive disorders in the year 2000. AB - BACKGROUND: The initial Global Burden of Disease study found that depression was the fourth leading cause of disease burden, accounting for 3.7% of total disability adjusted life years (DALYs) in the world in 1990. AIMS: To present the new estimates of depression burden for the year 2000. METHOD: DALYs for depressive disorders in each world region were calculated, based on new estimates of mortality, prevalence, incidence, average age at onset, duration and disability severity. RESULTS: Depression is the fourth leading cause of disease burden, accounting for 4.4% of total DALYs in the year 2000, and it causes the largest amount of non-fatal burden, accounting for almost 12% of all total years lived with disability worldwide. CONCLUSIONS: These data on the burden of depression worldwide represent a major public health problem that affects patients and society. PMID- 15123502 TI - Reducing the global burden of depression: population-level analysis of intervention cost-effectiveness in 14 world regions. AB - BACKGROUND: International evidence on the cost and effects of interventions for reducing the global burden of depression remain scarce. Aims To estimate the population-level cost-effectiveness of evidence-based depression interventions and their contribution towards reducing current burden. METHOD: Primary-care based depression interventions were modelled at the level of whole populations in 14 epidemiological subregions of the world. Total population-level costs (in international dollars or I$) and effectiveness (disability adjusted life years (DALYs) averted) were combined to form average and incremental cost-effectiveness ratios. RESULTS: Evaluated interventions have the potential to reduce the current burden of depression by 10-30%. Pharmacotherapy with older antidepressant drugs, with or without proactive collaborative care, are currently more cost-effective strategies than those using newer antidepressants, particularly in lower-income subregions. CONCLUSIONS: Even in resource-poor regions, each DALYaverted by efficient depression treatments in primary care costs less than 1 year of average per capita income, making such interventions a cost-effective use of health resources. However, current levels of burden can only be reduced significantly if there is a substantial increase substantial increase in treatment coverage. PMID- 15123503 TI - International variations in the outcome of schizophrenia and the prevalence of depression in relation to national dietary practices: an ecological analysis. AB - BACKGROUND: Dietary variations are known to predict the prevalence of physical illnesses such as diabetes and heart disease but the possible influence of diet on mental health has been neglected. AIMS: To explore dietary predictors of the outcome of schizophrenia and the prevalence of depression. METHOD: Ecological analysis of national dietary patterns in relation to international variations in outcome of schizophrenia and prevalence of depression. RESULTS: A higher national dietary intake of refined sugar and dairy products predicted a worse 2-year outcome of schizophrenia. A high national prevalence of depression was predicted by a low dietary intake of fish and seafood. CONCLUSIONS: The dietary predictors of outcome of schizophrenia and prevalence of depression are similar to those that predict illnesses such as coronary heart disease and diabetes, which are more common in people with mental health problems and in which nutritional approaches are widely recommended. Dietary intervention studies are indicated in schizophrenia and depression. PMID- 15123504 TI - Focal changes in brain energy and phospholipid metabolism in first-episode schizophrenia: 31P-MRS chemical shift imaging study at 4 Tesla. AB - BACKGROUND: Membrane phospholipid and high-energy abnormalities measured with phosphorus magnetic resonance spectroscopy ((31)P-MRS) have been reported in patients with schizophrenia in several brain regions. AIMS: Using improved imaging techniques, previously inaccessible brain regions were examined in patients with first-episode schizophrenia and healthy volunteers with 4.0 T (31)P MRS. METHOD: Brain spectra were collected in vivo from 15 patients with first episode schizophrenia and 15 healthy volunteers from 15 cm(3) effective voxels in the thalamus, cerebellum, hippocampus, anterior/posterior cingulate, prefrontal cortex and parieto-occipital cortex. RESULTS: People with first-episode schizophrenia showed increased levels of glycerophosphocholine in the anterior cingulate. Inorganic phosphate, phosphocreatine and adenosine triphosphate concentrations were also increased in the anterior cingulate in this group. CONCLUSIONS: The increased phosphodiester and high-energy phosphate levels in the anterior cingulate of brains of people with first-episode schizophrenia may indicate neural overactivity in this region during the early stages of the illness, resulting in increased excitotoxic neural membrane breakdown. PMID- 15123505 TI - Impact of child sexual abuse on mental health: prospective study in males and females. AB - BACKGROUND: The lack of prospective studies and data on male victims leaves major questions regarding associations between child sexual abuse and subsequent psychopathology. AIMS: To examine the association between child sexual abuse in both boys and girls and subsequent treatment for mental disorder using a prospective cohort design. METHOD: Children (n=1612; 1327 female) ascertained as sexually abused at the time had their histories of mental health treatment established by data linkage and compared with the general population of the same age over a specified period. RESULTS: Both male and female victims of abuse had significantly higher rates of psychiatric treatment during the study period than general population controls (12.4% v. 3.6%). Rates were higher for childhood mental disorders, personality disorders, anxiety disorders and major affective disorders, but not for schizophrenia. Male victims were significantly more likely to have had treatment than females (22.8% v.10.2%). CONCLUSIONS: This prospective study demonstrates an association between child sexual abuse validated at the time and a subsequent increase in rates of childhood and adult mental disorders. PMID- 15123506 TI - Somatic symptoms in depression: evaluation of their diagnostic weight in an African setting. AB - BACKGROUND: Somatic symptoms are extremely common features of depression and other mental disorders in African countries such as Nigeria, but their weight in the diagnosis of depression is not certain. AIM: To determine what weight should be assigned to these symptoms in comparison with other well-known symptoms in the diagnosis of depression. METHOD: A sample of 829 persons completed the Patient Health Questionnaire which was earlier modified by the inclusion of the somatic symptoms being studied. Using principal component analysis and a logistic regression model, the contributions of these symptoms in comparison with others were determined. RESULTS: Core depressive symptoms accounted for most of the total variance for depression. The somatic symptoms studied loaded separately from the core depressive symptoms and were not as good predictors of depression. A cognitive factor emerged as well as some somatic factors. CONCLUSIONS: Although somatic symptoms may be florid among patients with depression, they have considerably less weight than core depressive symptoms in the diagnosis of depression. The emerging cognitive factor could be similar to that described by previous authors. PMID- 15123507 TI - Ethnic variation among adolescent psychiatric in-patients with psychotic disorders. AB - BACKGROUND: There is strong evidence that the rates of psychiatric admission for psychosis in the UK are elevated for the Black adult population compared with the White population. Black adults also have a higher rate of involuntary psychiatric admissions. There have been no studies in this country investigating links between ethnic background and psychiatric admission in the adolescent population. AIMS: To investigate whether Black compared with White adolescents from London are overrepresented in psychiatric in-patient settings and whether they are more likely to be detained under the Mental Health Act 1983. METHOD: Cross-sectional survey of London adolescents aged 13-17 years, who were in-patients in psychiatric units. RESULTS: Adolescents from the Black group (Black African, Black Caribbean, Black British) were overrepresented among those admitted with a diagnosis of a psychotic disorder when compared with adolescents from the White group (White British, White Irish, White Other): odds ratio=3.7, 95% CI 2.0-6.7. They were also more likely to be detained on admission and more likely to be born outside the UK and more likely to be born outside the UK and have refugee background. CONCLUSIONS: The possible impact of various background factors influencing admission is discussed. PMID- 15123508 TI - Impact of compulsory community treatment on admission rates: survival analysis using linked mental health and offender databases. AB - BACKGROUND: There is controversy as to whether compulsory community treatment for psychiatric patients reduces hospital admission rates. AIMS: To examine whether community treatment orders (CTOs) reduce admission rates, using a two-stage design of matching and multivariate analyses to take into account socio demographic factors, clinical factors, case complexity and previous psychiatric and forensic history. METHOD: Survival analysis of CTO cases and controls from three linked Western Australian databases of health service use, involuntary treatment and forensic history. We used two control groups: one matched on demographic characteristics, diagnosis, past psychiatric history and treatment setting, and consecutive controls matched on date of discharge from in-patient care. RESULTS: We matched 265 CTO cases with 265 matched controls and 224 consecutive controls (total n=754). The CTO group had a significantly higher readmission rate: 72% v. 65% and 59% for the matched and consecutive controls (log-rank chi(2)=4.7, P=0.03). CTO placement, aboriginal ethnicity, younger age, personality disorder and previous health service use were associated with increased admission rates. CONCLUSIONS: Community treatment orders alone do not reduce admissions. PMID- 15123509 TI - Relationship between alcohol use disorders and suicidality in a psychiatric population: in-patient prevalence study. AB - BACKGROUND: Alcohol misuse is a risk factor in suicide and parasuicide. AIMS: To measure the prevalence of alcohol use disorders in a cohort of psychiatric admissions using the Alcohol Use Disorders Identification Test (AUDIT), and the relationship between the AUDIT score and suicidality. METHOD: Consecutive psychiatric admissions were interviewed with a lifestyle survey that included the AUDIT, and admission case notes were reviewed. RESULTS: Out of 200 subjects, 48.5% scored 8 or more (indicating hazardous or harmful alcohol use) and 22.5% scored 16 or more (indicating significant alcohol dependence) on the AUDIT. There were no significant gender differences. Alcohol misuse was strongly associated with suicidality. CONCLUSIONS: The AUDIT questionnaire should be incorporated into psychiatric assessments when risk of self-harm is being evaluated. Further research is warranted to examine the impact of interventions for alcohol use disorders in psychiatric settings on self-harm and suicidal ideation. PMID- 15123510 TI - Prematurity at birth and adolescent depressive disorder. AB - Association between prematurity/low birthweight and adolescent depressive disorder studied using a case-control design within a prospective cohort study of 2032 adolescents. Odds for depressive disorder were 11-fold (95% CI 2-62) higher for the premature/low-birthweight participants after regression adjustment for major confounding factors. For premature/low-birthweight females, cumulative rates of depressive disorder over 30 months were 15.2% (95% CI 11.1-20.5) v. 1.8% (95% CI 1.6-2.1) in those with normal deliveries. Physiological adaptations in utero before full term may be implicated causally in some cases of depression in adolescence. PMID- 15123511 TI - Computer-aided self-help for phobia/panic via internet at home: a pilot study. AB - In an open study, ten people with phobia or panic disorder who could not travel repeatedly to a therapist accessed a computer-aided exposure self-help system (FearFighter) at home on the internet with brief therapist support by telephone. They improved significantly, and their outcome and satisfaction resembled those in patients with similar disorders who used FearFighter in clinics with brief face-to-face therapist support. PMID- 15123512 TI - Mental health in the enlarged European Union: need for relevant public mental health action. PMID- 15123513 TI - Venlafaxine and SSRI remission data revisited. PMID- 15123514 TI - Need for medicine-based evidence in pharmacotherapy. PMID- 15123515 TI - Meanings and causes in ADHD. PMID- 15123516 TI - Commissioning conundrum for custodial care. PMID- 15123517 TI - A new name for the Journal? PMID- 15123518 TI - Integrated in-patient adolescent services. PMID- 15123521 TI - HMG-CoA reductase inhibitor simvastatin profoundly improves survival in a murine model of sepsis. AB - BACKGROUND: HMG-CoA reductase inhibitors, such as simvastatin, have been shown to exhibit pronounced immunomodulatory effects independent of lipid lowering but to date have not been used to treat severe inflammatory disease such as sepsis. We thus approached the question of whether treatment with simvastatin might improve cardiovascular function and survival in sepsis. METHODS AND RESULTS: Mice treated with simvastatin and rendered septic by cecal ligation and perforation (CLP) show a mean survival time close to 4 times the value found in untreated mice. This dramatic improvement is based on a complete preservation of cardiac function and hemodynamic status, which are severely impaired in untreated CLP mice [eg, 20 hours after CLP, cardiac output declined from 1.24+/-0.09 to 0.87+/-0.11 mL x min(-1) x g(-1) in untreated mice (P<0.005; n=12), while remaining unaltered (1.21+/-0.08 mL x min(-1) x g(-1) at baseline and 1.15+/-0.1 mL x min(-1) x g(-1) 20 hours after CLP, P=NS, n=12) in CLP mice treated with simvastatin]. Untreated CLP mice remained refractory to beta-stimulation, whereas the responsiveness to dobutamine was restored by treatment with simvastatin. Susceptibility of coronary flow to endothelial nitric oxide synthase (eNOS) stimulation by bradykinin was close to 3 times as pronounced in untreated CLP mice as in untreated sham operated mice, indicating a high level of eNOS activation secondary to sepsis. In addition, treatment with simvastatin reversed inflammatory alterations in CLP mice, namely, increased monocyte adhesion to endothelium. CONCLUSIONS: Simvastatin, which is well established in the treatment of lipid disorders and coronary artery disease, might have the additional potential of being an effective agent in sepsis treatment. PMID- 15123520 TI - Aldosterone administration to mice stimulates macrophage NADPH oxidase and increases atherosclerosis development: a possible role for angiotensin-converting enzyme and the receptors for angiotensin II and aldosterone. AB - BACKGROUND: The renin-angiotensin-aldosterone system is involved in the pathogenesis of atherosclerosis, partially because of its pro-oxidative properties. We questioned the effect and mechanisms of action of administration of aldosterone to apolipoprotein E-deficient (E(0)) mice on their macrophages and aorta oxidative status and the ability of pharmacological agents to block this effect. METHODS AND RESULTS: Aldosterone (0.2 to 6 microg. mouse(-1) x d(-1)) was administered to E(0) mice alone or in combination with eplerenone (200 mg x kg( 1) x d(-1)), ramipril (5 mg x kg(-1) x d(-1)), or losartan (25 mg x kg(-1) x d( 1)). Mouse aortic atherosclerotic lesion area and macrophage and aortic oxidative status were evaluated. Aldosterone administration enhanced the mouse atherosclerotic lesion area by 32%. Mouse peritoneal macrophages and aortic segments from aldosterone-treated mice exhibited increased superoxide anion formation by up to 155% and 69%, respectively, and this effect was probably mediated by NADPH oxidase activation, because increased translocation of its cytosolic component p47phox to the macrophage plasma membrane was observed. THP-1 macrophages incubated in vitro with aldosterone (10 micromol/L) exhibited a higher capacity to release superoxide ions by 110% and increased ability to oxidize LDL by 74% compared with control cells. Aldosterone administration enhanced mouse peritoneal macrophage ACE activity and mRNA expression by 2.3-fold and 2.4-fold, respectively. Only cotreatment of eplerenone with ramipril or losartan completely blocked the oxidative effects of aldosterone. CONCLUSIONS: Aldosterone administration to E(0) mice increased macrophage oxidative stress and atherosclerotic lesion development. Blocking of the mineralocorticoid receptor and inhibition of tissue ACE and/or the angiotensin receptor-1 reduced aldosterone deleterious pro-oxidative and proatherogenic effects. PMID- 15123522 TI - Optimal noninvasive assessment of left ventricular filling pressures: a comparison of tissue Doppler echocardiography and B-type natriuretic peptide in patients with pulmonary artery catheters. AB - BACKGROUND: Early transmitral velocity/tissue Doppler mitral annular early diastolic velocity (E/Ea) and B-type natriuretic peptide (BNP) have been correlated with left ventricular filling pressures, yet there are no data on how these 2 estimates of left ventricular filling pressures compare. METHODS AND RESULTS: Patients admitted to intensive care underwent simultaneous tissue Doppler echocardiography, BNP measurement, and pulmonary capillary wedge pressure (PCWP) determination. The ability of mitral E/Ea and BNP to predict PCWP >15 mm Hg was assessed. Fifty patients were studied. Ln BNP had a correlation of r=0.32 (P=0.02) with PCWP compared with r=0.69 (P<0.001) between E/Ea and PCWP. E/Ea >15 was the optimal cutoff to predict PCWP >15 mm Hg (sensitivity, 86%; specificity, 88%), whereas the optimal BNP cutoff was >300 pg/mL (sensitivity, 91%; specificity, 56%). The correlation between change in PCWP and change in E/Ea at 48 hours was r=0.87 (P=0.003) compared with r=-0.59 (P=0.39) for BNP. In the 36 patients with cardiac disease, E/Ea >15 (sensitivity, 92%; specificity, 91%) appeared more accurate than BNP >400 pg/mL (sensitivity, 92%; specificity, 51%), whereas in patients without cardiac disease, BNP (sensitivity, 81%; specificity, 83%) appeared more accurate than E/Ea >15 (sensitivity, 74%; specificity, 72%) for PCWP >15 mm Hg. CONCLUSIONS: In intensive care unit patients, mitral E/Ea has a better correlation than BNP with PCWP. Both BNP and mitral E/Ea have high sensitivity for PCWP >15 mm Hg; however, E/Ea appears more specific in this patient population. In patients without cardiac disease, BNP appears more accurate than E/Ea for PCWP >15 mm Hg, whereas E/Ea appears more accurate in patients with cardiac disease. PMID- 15123523 TI - Scintigraphic imaging of matrix metalloproteinase activity in the arterial wall in vivo. AB - BACKGROUND: Matrix metalloproteinases (MMPs) are enzymes involved in the proteolytic degradation of extracellular matrix. They play an important role in several disease processes, such as inflammation, cancer, and atherosclerosis. METHODS AND RESULTS: In this study, we have used the broad-spectrum MMP inhibitor CGS 27023A to develop the radioligand [123I]I-HO-CGS 27023A for in vivo imaging of MMP activity. Using this radioligand, we were able to specifically image MMP activity by scintigraphy in vivo in the MMP-rich vascular lesions that develop after carotid artery ligation and cholesterol-rich diet in apolipoprotein E deficient mice. These results were confirmed by gamma counting of lesional tissue (counts per minute per milligram). CONCLUSIONS: Imaging of MMP activity in vivo is feasible using radiolabeled MMP inhibitors. Additional studies are needed to test the potential of this approach as a novel noninvasive clinical diagnostic tool for the management of human MMP-related diseases. PMID- 15123524 TI - Impact of sirolimus-eluting stents on outcome in diabetic patients: a SIRIUS (SIRolImUS-coated Bx Velocity balloon-expandable stent in the treatment of patients with de novo coronary artery lesions) substudy. AB - BACKGROUND: Randomized clinical trials have shown that a sirolimus-eluting stent significantly reduces restenosis after percutaneous coronary revascularization. Diabetic patients are known to have a higher risk of restenosis compared with nondiabetic patients. The purpose of this analysis was to determine the impact of sirolimus-eluting stents on outcomes of diabetic compared with nondiabetic patients. METHODS AND RESULTS: The SIRIUS (SIRolImUS-coated Bx Velocity balloon expandable stent in the treatment of patients with de novo coronary artery lesions) trial is a randomized, double-blind study that compared sirolimus eluting and bare metal stent implantation in 1058 patients with de novo native coronary artery lesions. Diabetes mellitus was present in 279 (26%) patients (diabetes mellitus group, 131 patients received sirolimus-eluting stents and 148 patients received bare metal stents) and was absent in 778 patients (no-diabetes mellitus group, 402 patients received sirolimus-eluting stents and 376 patients received bare metal stents). At 270 days, target lesion revascularization was reduced in diabetic patients from 22.3% with bare metal stents to 6.9% with sirolimus-eluting stents (P<0.001) and in nondiabetic patients from 14.1% to 2.99% (P<0.001), respectively. Major adverse cardiac events were reduced in diabetic patients from 25% with bare metal stents to 9.2% with sirolimus-eluting stents (P<0.001) and from 16.5% to 6.5% (P<0.001) in nondiabetic patients, respectively. CONCLUSIONS: Implantation of sirolimus-eluting stents compared with bare metal stents in de novo coronary lesions reduces major adverse cardiac events in patients with and without diabetes mellitus. However, among patients receiving sirolimus-eluting stents, there remains a trend toward a higher frequency of repeat intervention in diabetic patients compared with nondiabetic patients, particularly in the insulin-requiring patients. PMID- 15123525 TI - Thioredoxin-interacting protein controls cardiac hypertrophy through regulation of thioredoxin activity. AB - BACKGROUND: Although cellular redox balance plays an important role in mechanically induced cardiac hypertrophy, the mechanisms of regulation are incompletely defined. Because thioredoxin is a major intracellular antioxidant and can also regulate redox-dependent transcription, we explored the role of thioredoxin activity in mechanically overloaded cardiomyocytes in vitro and in vivo. METHODS AND RESULTS: Overexpression of thioredoxin induced protein synthesis in cardiomyocytes (127+/-5% of controls, P<0.01). Overexpression of thioredoxin-interacting protein (Txnip), an endogenous thioredoxin inhibitor, reduced protein synthesis in response to mechanical strain (89+/-5% reduction, P<0.01), phenylephrine (80+/-3% reduction, P<0.01), or angiotensin II (80+/-4% reduction, P<0.01). In vivo, myocardial thioredoxin activity increased 3.5-fold compared with sham controls after transverse aortic constriction (P<0.01). Aortic constriction did not change thioredoxin expression but reduced Txnip expression by 40% (P<0.05). Gene transfer studies showed that cells that overexpress Txnip develop less hypertrophy after aortic constriction than control cells in the same animals (28.1+/-5.2% reduction versus noninfected cells, P<0.01). CONCLUSIONS: Thus, even though thioredoxin is an antioxidant, activation of thioredoxin participates in the development of pressure-overload cardiac hypertrophy, demonstrating the dual function of thioredoxin as both an antioxidant and a signaling protein. These results also support the emerging concept that the thioredoxin inhibitor Txnip is a critical regulator of biomechanical signaling. PMID- 15123526 TI - Innate immune recognition of invasive bacteria accelerates atherosclerosis in apolipoprotein E-deficient mice. AB - BACKGROUND: Infectious diseases have emerged as potential risk factors for cardiovascular disease (CVD). Epidemiological studies support a connection between periodontal disease, a chronic inflammatory disease of the supporting tissues of the teeth, and CVD. METHODS AND RESULTS: To directly test the connection between periodontal disease and atherosclerosis, apoE-/- mice were orally challenged with the periodontal disease pathogen Porphyromonas gingivalis or an invasion-impaired P gingivalis fimbriae-deficient mutant (FimA-). Both wild type P gingivalis and the FimA- mutant were detected in blood and aortic arch tissue of apoE-/- mice by PCR after challenge. ApoE-/- mice challenged with wild type P gingivalis presented with increased atherosclerotic plaque and expressed the innate immune response markers Toll-like receptor (TLR)-2 and TLR-4 in aortic tissue. Despite detection of the FimA- mutant in the blood and in aortic arch tissue, apoE-/- mice challenged with the FimA- mutant did not present with periodontal disease, upregulation of TLRs, or accelerated atherosclerosis. Furthermore, we demonstrate that immunization to control P gingivalis-elicited periodontal disease concomitantly prevents P gingivalis-accelerated atherosclerosis. CONCLUSIONS: We conclude that invasive P gingivalis accelerates atherosclerosis. PMID- 15123527 TI - Growth hormone replacement decreases plasma levels of matrix metalloproteinases (2 and 9) and vascular endothelial growth factor in growth hormone-deficient individuals. AB - BACKGROUND: Matrix metalloproteinases (MMP) are implicated in cardiovascular disease. Growth hormone (GH) deficiency is associated with increased cardiovascular mortality. We assessed whether GH replacement, in GH-deficient adults, has any effect on plasma levels of MMP-2 and MMP-9 and on vascular endothelial growth factor (VEGF), known to activate MMPs. METHODS AND RESULTS: The study comprised 66 GH-deficient adults, 37.8+/-14.7 years of age (37 female). Plasma MMP-2 and MMP-9, VEGF, and insulin-like growth factor-1 (IGF-1) were measured at baseline (V1), at 12 months (V2), and at 24 months of GH treatment (V3). IGF-1 levels rose under GH replacement (mean+/-SD): V1, 151.6+/-91.9 microg/mL; V2, 270.2+/-114.8 microg/mL; and V3, 266.2+/-109.8 (V1 versus V2; P<0.001: V2 versus V3; P=0.76). MMP-9 exhibited the most pronounced and sustained decline from 1248.0+/-651.1 ng/mL at V1, 949.2+/-457.7 ng/mL at V2, and 760.8+/ 386.1 ng/mL at V3 (P<0.001 at all time points). A similar pattern was detected for VEGF levels: 358.5+/-209.0 pg/mL at V1, 310.6+/-225.7 pg/mL at V2 (P<0.001), and 283.7+/-202.7 pg/mL at V3 (V2 versus V3; P=0.005). MMP-2 demonstrated a significant decline initially from V1 to V2 (1134.4+/-217.8 ng/mL versus 1074.5+/ 203.0 ng/mL, respectively; P=0.031), reaching a plateau at V3 (1072.3+/-220.2 ng/mL) (V2 versus V3; P=0.93). A negative relation existed between MMP-9 versus IGF-1 and MMP-2 versus IGF-1 (P<0.001 and P=0.007, respectively) as well as between VEGF and IGF-1 (P<0.001). CONCLUSIONS: These changes in MMPs and VEGF may contribute to the anticipated reduction in vascular mortality in hypopituitary adults receiving GH replacement. PMID- 15123528 TI - Prostaglandin E2 protects the heart from ischemia-reperfusion injury via its receptor subtype EP4. AB - BACKGROUND: In the heart with acute myocardial infarction, production of prostaglandin (PG) E2 increases significantly. In addition, several subtypes of PGE2 receptors (EPs) have been reported to be expressed in the heart. The role of PGE2 in cardiac ischemia-reperfusion (I/R) injury, however, remains unknown. We intended to clarify the role of PGE2 via EP4, an EP subtype, in I/R injury using mice lacking EP4 (EP4-/- mice). METHODS AND RESULTS: In murine cardiac ventricle, competitive reverse transcription-polymerase chain reaction revealed the highest expression level of EP4 mRNA among EP mRNAs. EP4-/- mice had larger infarct size than wild-type mice in a model of I/R; the left anterior descending coronary artery was occluded for 1 hour, followed by 24 hours of reperfusion. In addition, isolated EP4-/- hearts perfused according to the Langendorff technique had greater functional and biochemical derangements in response to I/R than wild-type hearts. In vitro, AE1-329, an EP4 agonist, raised cAMP concentration remarkably in noncardiomyocytes, whereas the action was weak in cardiomyocytes. When 4819 CD, another EP4 agonist, was administered 1 hour before coronary occlusion, it reduced infarct size significantly in wild-type mice. Notably, a similar cardioprotective effect was observed even when it was administered 50 minutes after coronary occlusion. CONCLUSIONS: Both endogenous PGE2 and an exogenous EP4 agonist protect the heart from I/R injury via EP4. The potent cardioprotective effects of 4819-CD suggest that the compound would be useful for treatment of acute myocardial infarction. PMID- 15123529 TI - Short-acting beta-adrenergic antagonist esmolol given at reperfusion improves survival after prolonged ventricular fibrillation. AB - BACKGROUND: High catecholamine concentrations are cytotoxic to cardiac myocytes. We hypothesized that myocardial interstitial catecholamine levels are greatly elevated immediately after long-duration ventricular fibrillation (VF), defibrillation, and reperfusion and that the short-acting beta-antagonist esmolol administered at reperfusion would protect against this catecholamine surge and improve survival. METHODS AND RESULTS: In part 1 of this study, catecholamines from myocardial interstitial fluid (ISF) and aortic and coronary sinus plasma were quantified by use of 3H-labeled radioenzymatic assay in 8 open-chest, anesthetized pigs. Eight minutes of electrically induced VF was followed by internal defibrillation and reperfusion. By 4 minutes of VF, ISF norepinephrine increased significantly, from 1.3+/-0.3 to 7.4+/-2.4 ng/mL. Epinephrine increased significantly, from 0.4+/-0.2 to 1.5+/-0.7 ng/mL. ISF norepinephrine and epinephrine peaked at 219.2+/-92.1 and 63.7+/-25.1 ng/mL after defibrillation and reperfusion and decreased significantly to 12.2+/-3.5 and 6.7+/-3.1 ng/mL 23 minutes after defibrillation. Transcardiac catecholamine changes were similar. In part 2, 8 minutes of VF was followed by external defibrillation in anesthetized, closed-chest pigs. Animals received 1.0 mg/kg esmolol (n=8) or saline (n=8) intravenously at the start of cardiopulmonary resuscitation (CPR). Advanced cardiac life support, including CPR and epinephrine, was delivered to both groups. Esmolol before reperfusion improved return of spontaneous circulation and 4-hour survival (7/8 versus 3/8 survivors, chi2 P<0.05). CONCLUSIONS: Transcardiac and ISF norepinephrine and epinephrine levels are briefly massively elevated after 8 minutes of VF, defibrillation, and reperfusion. A short-acting beta-antagonist administered immediately after defibrillation improves return of spontaneous circulation and 4-hour survival after this prolonged VF. PMID- 15123530 TI - Effect of obesity and insulin resistance on myocardial substrate metabolism and efficiency in young women. AB - BACKGROUND: Obesity is a risk factor for impaired cardiac performance, particularly in women. Animal studies suggest that alterations in myocardial fatty acid metabolism and efficiency in obesity can cause decreased cardiac performance. In the present study, we tested the hypothesis that myocardial fatty acid metabolism and efficiency are abnormal in obese women. METHODS AND RESULTS: We studied 31 young women (body mass index [BMI] 19 to 52 kg/m2); 19 were obese (BMI >30 kg/m2). Myocardial oxygen consumption (MVO2) and fatty acid uptake (MFAUp), utilization (MFAU), and oxidation (MFAO) were quantified by positron emission tomography. Cardiac work was measured by echocardiography, and efficiency was calculated as work/MVO2. BMI correlated with MVO2 (r=0.58, P=0.0006), MFAUp (r=0.42, P<0.05), and efficiency (r=-0.40, P<0.05). Insulin resistance, quantified by the glucose area under the curve (AUC) during an oral glucose tolerance test, correlated with MFAUp (r=0.55, P<0.005), MFAU (r=0.62, P<0.001), and MFAO (r=0.58, P<0.005). A multivariate, stepwise regression analysis showed that BMI was the only independent predictor of MVO2 and efficiency (P=0.0005 and P<0.05, respectively). Glucose AUC was the only independent predictor of MFAUp, MFAU, and MFAO (P<0.05, <0.005, and <0.005, respectively). CONCLUSIONS: In young women, obesity is a significant predictor of increased MVO2 and decreased efficiency, and insulin resistance is a robust predictor of MFAUp, MFAU, and MFAO. This increase in fatty acid metabolism and decrease in efficiency is concordant with observations made in experimental models of obesity. These metabolic changes may play a role in the pathogenesis of decreased cardiac performance in obese women. PMID- 15123531 TI - Delayed enhancement and T2-weighted cardiovascular magnetic resonance imaging differentiate acute from chronic myocardial infarction. AB - BACKGROUND: Delayed enhancement (DE) cardiovascular magnetic resonance (CMR) detects acute and chronic myocardial infarction (MI) by visualizing contrast media accumulation in infarcted segments. T2-weighted CMR depicts infarct-related myocardial edema as a marker of acute but not chronic myocardial injury. We investigated the clinical utility of an approach combining both techniques to differentiate acute from chronic MI. METHODS AND RESULTS: Seventy-three MI patients were studied in 2 groups. Group A consisted of 15 acute MI patients who were studied twice, on day 1 and 3 months after MI. In group B, 58 patients with acute or chronic MI underwent 1 CMR scan. T2-weighted and DE images of matched slices were acquired on a 1.5-T system. In group A, quantitative segmental and region of interest-based analyses were performed to observe signal changes between the acute and chronic phases. In group B, T2-weighted and DE images were examined visually by 2 blinded observers for the presence or absence of hyperintense areas in corresponding segments. For infarct localization, coronary angiography and/or ECG changes served as the reference standard. In group A, the contrast-to-noise ratio on T2-weighted images dropped in the infarcted segments from 2.7+/-1.1 on day 1 to 0.1+/-1.2 after 3 months (P<0.0001). There was no significant change in contrast-to-noise ratio in DE images (1.9+/-1.5 versus 1.3+/-1.0; P=NS). The qualitative assessment of T2-weighted and DE images in group B yielded a specificity of 96% to differentiate acute from chronic lesions. CONCLUSIONS: An imaging approach combining DE and T2-weighted CMR accurately differentiates acute from chronic MI. PMID- 15123532 TI - Peroxisome proliferator-activated receptor ligand bezafibrate for prevention of type 2 diabetes mellitus in patients with coronary artery disease. AB - BACKGROUND: Recent studies have shown that type 2 diabetes is preventable by both lifestyle interventions and medications that influence primary glucose metabolism. Whether pharmacological interventions that influence primary lipid metabolism can also delay development of type 2 diabetes is unknown. The goal of this study was to evaluate the effect of the peroxisome proliferator-activated receptor ligand bezafibrate on the progression of impaired fasting glucose phase to type 2 diabetes in patients with coronary artery disease over a 6.2-year follow-up period. METHODS AND RESULTS: The study sample comprised 303 nondiabetic patients 42 to 74 years of age with a fasting blood glucose level of 110 to 125 mg/dL (6.1 to 6.9 mmol/L). The patients received either 400 mg bezafibrate retard (156 patients) or placebo (147 patients) once a day. No patients were using statins, and use of ACE inhibitors, which also reduce diabetes incidence, was relatively low. During follow-up, development of new-onset diabetes was recorded in 146 patients: in 80 (54.4%) from the placebo group and 66 (42.3%) from the bezafibrate group (P=0.04). The mean time until onset of new diabetes was significantly delayed in patients on bezafibrate compared with patients on placebo: 4.6+/-2.3 versus 3.8+/-2.6 years (P=0.004). Multivariate analysis identified bezafibrate treatment as an independent predictor of reduced risk of new diabetes development (hazard ratio, 0.70; 95% CI, 0.49 to 0.99). Other significant variables associated with future overt type 2 diabetes in patients with impaired fasting glucose were total cholesterol level (hazard ratio, 1.22; 95% CI 1.0 to 1.51) and body mass index (hazard ratio, 1.10; 95% CI, 1.05 to 1.16). CONCLUSIONS: Bezafibrate reduces the incidence and delays the onset of type 2 diabetes in patients with impaired fasting glucose. Whether the combination of bezafibrate with other recommended drugs for secondary prevention (statins and ACE inhibitors) would be as efficacious as suggested by our results remains to be determined. PMID- 15123533 TI - Six- and twelve-month results from first human experience using everolimus eluting stents with bioabsorbable polymer. AB - BACKGROUND: Everolimus, an active immunosuppressive and antiproliferative agent of the same family as sirolimus (rapamycin), has demonstrated significant reduction of neointimal proliferation in animal studies. The First Use To Underscore restenosis Reduction with Everolimus (FUTURE) I trial was the first in human experience to evaluate the safety and efficacy of everolimus-eluting stents (EES), coated with a bioabsorbable polymer, compared with bare metal stents (BMS). METHODS AND RESULTS: FUTURE I was a prospective, single-blind, randomized trial that enrolled 42 patients with de novo coronary lesions (EES 27, BMS 15). Patient and lesion characteristics were comparable between the groups. Major adverse cardiac event rates were low at 30 days and 6 months, without any early or late stent thrombosis for either group (P=NS). Between 6 and 12 months, there were no additional reports of major adverse cardiac events. The 6-month angiographic in-stent restenosis rate was 0% versus 9.1% (1 patient) (P=NS), with an associated late loss of 0.11 mm versus 0.85 mm (P<0.001), and the in-segment restenosis rate was 4% (1 patient) and 9.1% (1 patient) (P=NS) for EES and BMS, respectively. Intravascular ultrasound analysis revealed a significant reduction of percent neointimal volume in EES compared with BMS (2.9+/-1.9 mm3/mm versus 22.4+/-9.4 mm3/mm, P<0.001). There was no late stent malapposition in either group. The safety and efficacy of the EES appeared to be sustained at 12 months. CONCLUSIONS: In this initial clinical experience, EES with bioabsorbable polymer demonstrated a safe and efficacious method to reduce in-stent neointimal hyperplasia and restenosis. PMID- 15123534 TI - Neurological events during long-term mechanical circulatory support for heart failure: the Randomized Evaluation of Mechanical Assistance for the Treatment of Congestive Heart Failure (REMATCH) experience. AB - BACKGROUND: Progression of heart failure can lead to cardiac transplantation, but when patients are ineligible, long-term mechanical circulatory support may improve survival. The REMATCH trial showed that left ventricular assist devices (LVADs) prolonged survival in patients with end-stage disease, but with a significant number of adverse events. We report on the neurological outcomes in the REMATCH trial. METHODS AND RESULTS: We examined new neurological events in the 129 patients randomized to either LVAD placement (n=68) or medical management (n=61), classified as stroke, transient ischemic attack, toxic-metabolic encephalopathy, and other. There were 46 neurological events: 42 in 30 LVAD patients and 4 in 4 patients in the medical arm (chi2, 30/68 versus 4/61, P<0.001). Sixteen percent of the LVAD patients had a stroke, with a rate of 0.19 per year (95% CI, 0.10 to 0.33), many occurring in the postoperative period. The stroke rate in the medical arm was 0.052. A Kaplan-Meier survival analysis showed a 44% reduction in the risk of stroke or death in the LVAD group versus the optimal medical group (P=0.002). The mean interval from implantation to stroke was 221.8 days (+/-70.4 days). History of stroke, age, and sepsis were not stroke risk factors in the LVAD group. CONCLUSIONS: Fewer than half of the patients in the LVAD group had a neurological event, and there were few neurological deaths. Survival analysis combining stroke or death demonstrated a significant benefit for long-term circulatory support with an LVAD over medical therapy. Future trials will need to address prospectively all neurological outcomes, including neurocognitive function, and the role of long-term neuroprotection. PMID- 15123535 TI - Acceleration of the healing process and myocardial regeneration may be important as a mechanism of improvement of cardiac function and remodeling by postinfarction granulocyte colony-stimulating factor treatment. AB - BACKGROUND: We investigated whether the improvement of cardiac function and remodeling after myocardial infarction (MI) by granulocyte colony-stimulating factor (G-CSF) relates to acceleration of the healing process, in addition to myocardial regeneration. METHODS AND RESULTS: In a 30-minute coronary occlusion and reperfusion rabbit model, saline (S) or 10 microg x kg(-1) x d(-1) of human recombinant G-CSF (G) was injected subcutaneously from 1 to 5 days after MI. Smaller left ventricular (LV) dimension, increased LV ejection fraction, and thicker infarct-LV wall were seen in G at 3 months after MI. At 2, 7, and 14 days and 3 months after MI, necrotic tissue areas were 14.2+/-1.5/13.4+/-1.1, 0.4+/ 0.1/1.8+/-0.5*, 0/0, and 0/0 mm2 x slice(-1) x kg(-1), granulation areas 0/0, 4.0+/-0.7/8.5+/-1.0*, 3.9+/-0.8/5.7+/-0.7,* and 0/0 mm2 x slice(-1) x kg(-1), and scar areas 0/0, 0/0, 0/0, and 4.2+/-0.5/7.9+/-0.9* mm2 x slice(-1) x kg(-1) in G and S, respectively (*P<0.05, G versus S). Clear increases of macrophages and of matrix metalloproteinases (MMP) 1 and 9 were seen in G at 7 days after MI. This suggests that G accelerates absorption of necrotic tissues via increase of macrophages and reduces granulation and scar tissues via expression of MMPs. Meanwhile, surviving myocardial tissue areas within the risk areas were significantly increased in G despite there being no difference in LV weight, LV wall area, or cardiomyocyte size between G and S. Confocal microscopy revealed significant increases of cardiomyocytes with positive 3,3,3',3' tetramethylindocarbocyanine perchlorate and positive troponin I in G, suggesting enhanced myocardial regeneration by G. CONCLUSIONS: The acceleration of the healing process and myocardial regeneration may play an important role for the beneficial effect of post-MI G-CSF treatment. PMID- 15123536 TI - De novo testosterone production in luteinizing hormone receptor knockout mice after transplantation of leydig stem cells. AB - Mesenchymal stem cells or Leydig cell progenitors are rare and difficult to isolate from adult testes. The property of differential efflux of Hoechst 33342 dye by the multi-drug-like transporter enriches murine hematopoietic stem cells from bone marrow. Our work on testicular cell transplantation suggests that the "Hoechst dim" side population (SP) also contains Leydig stem cells or progenitors that proliferate and differentiate into mature functional Leydig cells. We harvested testicular cells from cryptorchid ROSA26 mice, stained them with Hoechst dye, and isolated the cell population that excludes the dye using flow cytometry. Mice with targeted deletion of the LH receptor (LHR) gene were used as the recipients of the transplanted cells. These mice are hypogonadal and infertile. Both testicular SP and non-SP cells were transplanted into the interstitium of the LHR knockout recipients' testes. Serial serum testosterone assays revealed a significant increase in the circulating testosterone levels and restoration of spermatogenesis in the LHR-knockout recipients transplanted with the SP cells compared with that of those transplanted with non-SP. A SP cell concentration- and time-dependent increase in circulating testosterone was observed. This demonstrates the successful transplantation of functional putative Leydig stem cells into a hypogonadal recipient. The increase in testosterone concentration indicates the de novo synthesis of androgen by the transplanted SP cells. This method offers a novel technique to isolate Leydig stem cells and to study Leydig cell development. PMID- 15123537 TI - Differential roles for cholecystokinin a receptors in energy balance in rats and mice. AB - Although cholecystokinin A (CCK-A) receptors (CCK-AR) mediate the feeding inhibitory actions of CCK in both rats and mice, the absence of CCK-AR results in species-specific phenotypes. The lack of CCK-AR in Otsuka Long-Evans Tokushima fatty (OLETF) rats results in hyperphagia and obesity. We have suggested that demonstrated increases in meal size and elevated levels of dorsomedial hypothalamic (DMH) neuropeptide Y (NPY) gene expression may contribute to this phenotype. In contrast to OLETF rats, CCK-AR(-/-) mice have normal total daily food intake and do not develop obesity. To assess the basis underlying the different phenotypes in rats and mice lacking CCK-AR, we characterized meal patterns in CCK-AR(-/-) mice and determined whether CCK-AR(-/-) mice exhibited an alteration in DMH NPY gene expression. We demonstrate that although CCK-AR(-/-) mice show a similar dysregulation in meal size as OLETF rats, they do not have an elevation in DMH NPY mRNA expression levels. In fact, intact mice have no CCK-AR in the DMH. Furthermore, in intact rats, NPY and CCK-AR are colocalized in DMH neurons, and parenchymal injection of CCK into the DMH reduces food intake and down-regulates DMH NPY mRNA expression. These results suggest that although CCK AR plays a role in the mediation of CCK actions in the control of meal size in both rats and mice, CCK-AR seems to contribute to modulating DMH NPY levels only in rats. The deficit in CCK's action in the control of DMH NPY gene expression may play a major role in the obese phenotype in OLETF rats. PMID- 15123538 TI - Naked deoxyribonucleic acid vaccination induces recognition of diverse thyroid peroxidase T cell epitopes. AB - Recently, we observed that vaccination of BALB/c mice with thyroid peroxidase (TPO)-DNA in a plasmid is highly effective at inducing antibodies that interact with the immunodominant region recognized by human autoantibodies. We have now analyzed the TPO epitopes recognized by memory T cells in these animals. Splenocytes from TPO-DNA (not control DNA)-vaccinated mice responded to TPO protein antigen, as measured by interferon-gamma production. As a group, TPO immunized mice recognized 35 of 55 overlapping synthetic peptides that encompass the 814-amino acid TPO ectodomain. In individual mice, between five and 10 peptides induced splenocyte responses. Two T cell epitopes were immunodominant, one of which is also recognized by patients with autoimmune thyroid disease. To explore a potential correlation between T and B cell epitopes, we analyzed serum TPO antibody epitopic fingerprints. No relationship was evident. However, the number of T cell epitopes recognized by individual mice was inversely proportional to recognition of an antibody epitopic subdomain. The diversity of TPO T cell epitopes is in striking contrast to the restricted number of TSH receptor (TSHR) peptides (four of 29) recognized by T cells, as is the paucity of antibodies in the same strain of mice vaccinated with TSHR-DNA. In conclusion, our data highlight differences for both antibody and T cell epitopic recognition in TPO- vs. TSHR-DNA-immunized BALB/c mice. These findings provide insight into mechanisms that may be involved in spontaneous immune responses to two major thyroid autoantigens in humans. PMID- 15123539 TI - Radial artery bypass grafts have an increased occurrence of angiographically severe stenosis and occlusion compared with left internal mammary arteries and saphenous vein grafts. AB - BACKGROUND: The radial artery has been increasingly used in CABG. However, angiographic outcome data have been limited. METHODS AND RESULTS: We reviewed all coronary angiography procedures from February 1996 to October 2001 and selected patients with a radial artery bypass graft. Angiographic outcomes were divided into groups as (1) occluded, (2) severe disease (> or =70% stenosis, or string sign), or (3) patent (<70% stenosis). Multivariable analyses determined predictors of severe disease or occlusion. A total of 310 patients had a radial artery graft. Mean follow-up after coronary artery bypass grafting was 565+/-511 days. Radial artery grafts had a patency rate of 51.3%, which was significantly lower than that for left internal mammary arteries (90.3%, P<0.0001) or saphenous vein grafts (64.0%, P=0.0016). Radial artery grafts had an occlusion rate of 33.7%, compared with 4.8% for left internal mammary arteries (P<0.0001), and had a severe stenosis rate of 15.1%, compared with 5.9% for saphenous vein grafts (P=0.0003) and 4.8% for left internal mammary arteries (P<0.0001). Women had a worse overall radial artery patency rate than men (38.9% versus 56.1%, P=0.025). A radial artery graft was the most powerful multivariable predictor of severe stenosis or occlusion (chi2=28.87, P<0.0001). Because of diseased radial artery grafts, 58 patients required subsequent percutaneous intervention, and 26 patients required repeat CABG. CONCLUSIONS: In patients predominantly presenting with signs and symptoms of myocardial ischemia after CABG, radial artery grafts have lower patency rates than left internal mammary artery and saphenous vein grafts. Selective use of the radial artery is warranted, particularly in women. PMID- 15123540 TI - Images in cardiovascular medicine. Coronary uptake of gallium-67 citrate after implantation of sirolimus-eluting stent. PMID- 15123541 TI - Images in cardiovascular medicine. Massive left ventricular aneurysm. PMID- 15123542 TI - Rofecoxib use increases acute myocardial infarction risk. PMID- 15123543 TI - Mechanisms of channel gating of the ligand-gated ion channel superfamily inferred from protein structure. AB - The nicotinic-like ligand-gated ion channel superfamily consists of a group of structurally related receptors that activate an ion channel after the binding of extracellular ligand. The recent publications of the crystal structure of an acetylcholine binding protein and a refined electron micrograph structure of the membrane-bound segment of an acetylcholine receptor have led to insights into the molecular determinants of receptor function. Although the structures confirmed much biochemical and electrophysiological data obtained about the receptors, they also provide opportunities to study further the mechanisms that allow channel activation stimulated by ligand-binding. Here we review the mechanisms of channel gating that have been elucidated by information gained from the structures of the acetylcholine binding protein and membrane-bound segment of the acetylcholine receptor. PMID- 15123544 TI - Role of sex, gonadectomy and sex hormones in the development of nitric oxide inhibition-induced hypertension. AB - In this study we have evaluated the influence of sex, gonadectomy and sex hormones on the development of L-NAME-induced hypertension in the rat, focusing our investigation on blood pressure (BP), plasma renin activity (PRA), cardiac hypertrophy and proteinuria. Three experiments were performed to investigate: (i) the influence of sex on the development of L-NAME-induced hypertension; (ii) the effects of gonadectomy on the dimorphism of L-NAME-induced hypertension; and (iii) the effects of testosterone in ovariectomized female and of 17beta oestradiol in orchidectomized male rats. Male L-NAME-treated rats had higher BP values than females. Orchidectomy of L-NAME-treated rats reduced BP to the levels of females and ovariectomy did not affect hypertension in females. Oestrogenized and orchidectomized males had a BP level similar to intact female L-NAME-treated rats. However, androgenization and ovariectomy did not change BP in female L-NAME treated rats. PRA was greater in intact male L-NAME hypertensive rats than in female rats, and gonadectomy protected against the increase in PRA such that PRA was similar among all the groups. Intact female hypertensive rats showed significantly greater ventricular hypertrophy compared with male hypertensive rats. Male L-NAME hypertensive rats had increased proteinuria that was not present in female rats. Moreover, testosterone increased proteinuria in males and females regardless of the BP level. Male L-NAME-treated rats developed higher BP, PRA and proteinuria than female rats, but were more resistant to the development of cardiac hypertrophy. The higher PRA of male L-NAME-treated rats might be involved in the sex-dependent dimorphism of this model of hypertension. PMID- 15123545 TI - The electrophysiological and mechanical effects of 2,3-butane-dione monoxime and cytochalasin-D in the Langendorff perfused rabbit heart. AB - Procedures that reduce contraction are used to facilitate optical measurements of membrane potential, but it is unclear to what extent they affect the excitability of the heart. This study has examined the electrophysiological consequences of a range of extracellular [Ca2+] (0.7-2.5 mmol l(-1)), 2,3-butane-dione monoxime (BDM; 1-20 mmol l(-1)) and cytochalasin-D (Cyto-D; 1-5 micromol l(-1)). METHODS: Monophasic action potentials (MAPs) were recorded from the basal epicardial surface of the left ventricle of isolated rabbit hearts. Conduction delay (CD) and time to 90% repolarisation of the monophasic action potential (MAPD90) were measured. The effects of BDM and Cyto-D on restitution were studied at a [Ca2+] of 1.9 mmol l(-1). Restitution curves for MAPD90 were generated using a standard S1-S2 protocol. RESULTS: All manoeuvres decreased left ventricular developed pressure (LVDP): 0.7 mmol l(-1) Ca2+ to 74.0 +/- 6.1%, 20 mmol l(-1) BDM to 4.5 +/- 1.0%, and 5 micromol l(-1) Cyto-D to 12.8 +/- 3.5% of control value. CD decreased from a control value (33.3 +/- 1.0 ms, n= 16) to 93.0 +/- 2.2% in 0.7 mmol l(-1) Ca2+, but increased to 133.7 +/- 10.5% in 20 mmol l(-1) BDM and 127.4 +/- 10.6% in 5 micromol l(-1) Cyto-D. At 350 ms pacing cycle length, MAPD90 (control = 119.6 +/- 1.7 ms n= 16) was prolonged by reduced extracellular [Ca2+]. BDM had no effects on MAPD90 at control pacing rates. Cyto-D caused a significant prolongation (to 115.0 +/- 3.0% of control, n= 6) at the highest concentration studied (5 micromol l(-1)). Both BDM (20 mmol l(-1)) and Cyto-D (3 micromol l( 1)) flattened the restitution curves but neither agent altered maximum MAPD90. CONCLUSIONS: Extracellular [Ca2+] of 1.9 mmol l(-1) in conjunction with a moderate dose of Cyto-D (3 micromol l(-1)) reduced contractility with minimal effects on action potential duration and conduction at a fixed pacing cycle length. However, both BDM and Cyto-D had pronounced effects on electrical restitution. PMID- 15123546 TI - Volume regulation is defective in renal proximal tubule cells isolated from KCNE1 knockout mice. AB - The membrane protein KCNE1 has been implicated in cell volume regulation. Using a knockout mouse model, this study examined the role of KCNE1 in regulatory volume decrease (RVD) in freshly isolated renal proximal tubule cells. Cell diameter was measured using an optical technique in response to hypotonic shock and stimulation of Na(+)-alanine cotransport in cells isolated from wild-type and KCNE1 knockout mice. In HEPES buffered solutions 64% of wild-type and 56% of knockout cells demonstrated RVD. In HCO3- buffered solutions 100% of the wild type cells showed RVD, while in the knockout cells the proportion of cells displaying RVD remained unchanged. RVD in the knockout cells was rescued by valinomycin, a K+ ionophore. In wild-type HCO3- dependent cells the K+ channel inhibitors barium and clofilium inhibited RVD. These data suggest that mouse renal proximal tubule is comprised of two cell populations. One cell population is capable of RVD in the absence of HCO3-, whereas RVD in the other cell population has an absolute requirement for HCO3-. The HCO3- dependent RVD requires the normal expression of KCNE1. PMID- 15123547 TI - Application of red laser video-rate scanning confocal microscopy to in vivo assessment of tubular function in the rat: selective action of diuretics on tubular diameter. AB - This study examined the use of a red laser illuminated, video-rate scanning confocal reflection microscopy (VRSCM) system, with improved structural and functional imaging at high temporal resolution, to visualize physiological changes in the kidney in response to pharmacological stimuli. We applied VRSCM to superficial nephrons in vivo and measured temporal changes in the diameter of proximal and/or distal tubular segments in response to the administration of three major classes of diuretics with known selective actions at specific nephron sites. Mannitol caused measurable increases in both proximal and distal tubular diameter, whereas frusemide and hydrochlorothiazide caused dilation of the distal tubules only. The findings indicate that VRSCM is capable of detecting and quantifying predicted dynamic changes in renal tubular diameter. PMID- 15123548 TI - Glutamate and tachykinin receptors in central sensitization of withdrawal reflexes in the decerebrated rabbit. AB - This study assessed the involvement of NMDA and group I metabotropic glutamate receptors, and tachykinin NK1 and NK3 receptors, in central sensitization of withdrawal reflexes in the decerebrated rabbit. Reflexes evoked in the ankle flexor tibialis anterior and the knee flexor semitendinosus by electrical stimulation at the base of the toes were enhanced for 29-63 min after application of 20% mustard oil to the tips of the toes. Selective antagonists of mGlu1, mGlu5, NMDA and NR2B-subunit-containing NMDA glutamate receptors, as well as NK1, and NK3 receptors, and a non-selective blocker of all tachykinin receptors, were assessed for their effects on the magnitude and duration of the increase in reflexes induced by mustard oil. Dizocilpine, an antagonist of all NMDA receptors (1 mg intrathecal) abolished facilitation of tibialis anterior reflexes and significantly reduced the magnitude and duration of increase of the semitendinosus response. The NR2B-subtype selective antagonist CP-101,606 decreased the magnitude of facilitation of both reflexes but had no effect on duration of enhancement. Selective antagonists for the mGlu1 (CPCCOEt, 1-3 mg intrathecal), mGlu5 (MPEP, 0.2-1 mg intrathecal), NK1 (L-733,060, 0.3 mg intrathecal) or NK3 (SR 142,801, 1 mg kg(-1) i.v.) receptors had no effect on the amplitude or duration of sensitization. However, the non-selective tachykinin receptor blocker ZD-6021 (0.3 mg intrathecal) reduced the amplitude but not the duration of sensitization in the flexor reflexes. Combination of ZD-6021 with CP 101,606 (doses as above) decreased both aspects of the sensitization response. Dizocilpine reduced reflexes evoked from the heel per se, and dizocilpine, CP 101,606 and ZD-6021 reduced arterial blood pressure. Otherwise the drugs used had no effects on baseline variables. The present data confirm the importance of NMDA receptors as a critical part of the process of central sensitization, provide no evidence for a role of metabotropic glutamate receptors, and show that simultaneous blockade of all tachykinin receptors is required to reveal their role in hyperalgesia. The data further indicate that a combined pharmacological approach offers a potential way forward for the development of new antihyperalgesic agents. PMID- 15123549 TI - Heart rate and heart rate variability in chicken embryos at the end of incubation. AB - Our immediate goal was to study heart rate variability (HRV) in chicken embryos in the egg. Instantaneous heart rate data were needed for this purpose, and accordingly an ECG recording method in the egg was developed. The aim of this work was to test the hypothesis that autonomic nervous cardiac modulation, as shown from HRV parameters, is present at the end of development and that it reaches a constant value during the last days of incubation. Embryonic chicken heart rate was obtained at the final incubation period (days 19 and 20) from ECG recordings. Tachograms were computed and time- and frequency-domain indices of HRV were determined. No significant differences were found between HRV indices from day 19 and day 20. The power spectra extended in two frequency bands with centre frequency around 0.6-0.7 Hz (low frequency (LF) component), and another around 1.2-1.5 Hz (high frequency (HF) component); the latter was shown to reflect respiratory sinus arrhythmia. A relation between mean RR interval and some HRV parameters (rMSSD, pNN5 and HF power) was shown. HRV results obtained from embryonic chickens, showed the presence of modulation of cardiovascular function by the autonomic nervous system. The results suggested that sympathetic and parasympathetic activities have already reached a constant level at day 19 of incubation. High frequency oscillations (0.78-2.5 Hz) were detected and are considered to reflect respiratory sinus arrhythmia. PMID- 15123550 TI - Phenotypic differences in cholinergic responses of distal colonic epithelium. AB - The Flinders sensitive line (FSL) rats exhibit an increased cholinergic responsiveness in vivo when compared to their counterparts, the Flinders resistant line (FRL) rats. The functional consequences of this phenotypic difference on colonic mucosal function are not known. We sought to determine whether isolated distal colonic mucosa from the two strains exhibit differential responses to cholinergic agonists. The responses of the distal colonic mucosa from two lines of rats to carbachol were compared by recording changes in short circuit current. The ion movements associated with these changes were assessed by flux analysis of the radiotracers, 22Na and 36Cl. The anticipated hyper responsiveness to cholinergic stimulation in FSL rats was not seen. Carbachol responses were significantly enhanced by indomethacin pretreatment only in FRL rats. Tetrodotoxin (TTX) pretreatment significantly reduced responses to carbachol in FSL rats at all concentrations tested, though this was only seen with lower concentrations in FRL rats. Flux analysis indicated that both lines absorbed Na+ and Cl- under basal conditions and that a significant residual flux was present. Stimulation with carbachol led to significant reductions in net Na+ and Cl- fluxes in both lines. The changes in net Na+ and Cl- flux in both lines stem largely from a decrease in mucosal to serosal fluxes of both ions with an increase in serosal to mucosal flux of Cl-. The striking difference is the significant reduction in residual flux seen only in FRL rats. Indomethacin pretreatment abolished the changes in residual flux seen in FRL rats. Thus the responses to carbachol in these rats had at least three components: (a) a direct effect on the transporting colonocyte, (b) an indirect effect mediated by an arachidonic acid metabolite, and (c) another indirect effect involving a neurotransmitter. The relative contributions of each of these components were different in the two lines. PMID- 15123551 TI - Nitric oxide-dependent protein synthesis in parotid and submandibular glands of anaesthetized rats upon sympathetic stimulation or isoprenaline administration. AB - In anaesthetized female rats, the beta-adrenoceptor agonist isoprenaline was intravenously infused (20 microg kg(-1) min(-1)) for 30 min or the ascending cervical sympathetic nerve trunk was intermittently stimulated (50 Hz, 1 s every tenth second) on one side for 30 min. The incorporation of [3H]leucine into trichloroacetic acid (TCA)-insoluble material was used as an index of protein synthesis. In response to isoprenaline, the [3H]leucine incorporation increased by 79% in the parotid glands and by 82% in the submandibular glands. The neuronal type NO-synthase inhibitor N-PLA, reduced (P < 0.001) this response to 26% and 20%, respectively. Sympathetic stimulation under alpha-adrenoceptor blockade increased the [3H]leucine incorporation by 192% in the parotid glands and by 35% in the submandibular glands. N-PLA reduced the corresponding percentage figures to 86% (P < 0.01) and 8% (P < 0.05). When tested in the parotid glands, the non selective NO-synthase inhibitor L-NAME reduced (P < 0.01) the nerve-evoked response to 91%. The increase in [3H]leucine incorporation in response to sympathetic stimulation under beta-adrenoceptor blockade was not affected by N PLA in the parotid (139% versus 144%) and submandibular glands (39% versus 34%). In non-stimulated glands, the [3H]leucine incorporation was not influenced by the NO-synthase inhibitors. In conclusion, beta-adrenoceptor mediated salivary gland protein synthesis is largely dependent on NO generation by neuronal type NO synthase, most likely of parenchymal origin. PMID- 15123552 TI - Delayed shrinkage triggered by the Na+-K+ pump in terbutaline-stimulated rat alveolar type II cells. AB - Terbutaline (10 microm) induced a triphasic volume change in alveolar type II (AT II) cells: an initial shrinkage (initial phase) followed by cell swelling (second phase) and a gradual shrinkage (third phase). The present study demonstrated that the initial and the third phases are evoked by the activation of K+ and Cl- channels and the second phase is evoked by the activation of Na+ and Cl- channels. Ouabain blocked the third phase, although it did not block the initial and second phases. This suggests that the third phase is triggered by the Na+-K+ pump. Tetraethylammonium (TEA, a K+ channel blocker) decreased the volume of AT II cells and enhanced the terbutaline-stimulated third phase, although quinidine, another K+ channel blocker, increased the volume of AT-II cells. The TEA-induced cell shrinkage was inhibited by ouabain, suggesting that TEA increases Na+-K+ pump activity. Ba2+, 2,3-diaminopyridine and a high [K+]o (30 mm) similarly decreased the volume of AT-II cells. These findings suggest that depolarization induced by TEA increases Na+-K+ pump activity, which increases [K+]i. This [K+]i increase, in turn, hyperpolarizes membrane potential. Valinomycin (a K+ ionophore), which induces hyperpolarization, decreased the volume of AT-II cells and enhanced the third phase in these cells. In conclusion, in terbutaline stimulated AT-II cells, an increase in Na+-K+ pump activity hyperpolarizes the membrane potential and triggers the third phase by switching net ion transport from NaCl entry to KCl release. PMID- 15123553 TI - The role of endothelial cell Ca2+ store release in the regulation of microvascular permeability in vivo. AB - Microvascular permeability is regulated by changes in intracellular calcium concentration. The mechanism by which this increase in calcium determines permeability under normal conditions and during stimulation with agonists remains to be elucidated. In order to determine whether calcium release from intracellular stores could contribute towards the regulation of vascular permeability, hydraulic conductivity (Lp) was measured in frog mesenteric microvessels during stimulation of the endothelial cells of these vessels with agonists that release calcium from the intracellular stores. ATP (which acts through activation of inositol 1,4,5-trisphosphate (IP3) receptors) increased Lp in the absence of calcium influx across the plasma membrane 2.3 +/- 0.3 fold (mean +/- s.e.m., P < 0.01, n = 8), which was less than the increase in the presence of calcium influx (3.1 +/- 1.1 fold). Caffeine (which acts through activation of ryanodine receptors) also increased Lp in the absence of calcium influx across the plasma membrane 3.8 +/- 1.0 fold (P < 0.01, n = 9), but by at least as much as it does in the presence of calcium influx (2.8 +/- 0.5 fold). It is surprising that there was a strong positive correlation between the size of the response during store release and the baseline permeability (r = 0.91 for ATP, r = 0.75 for caffeine). This suggests that the filling state of the stores may regulate the baseline permeability of the microvessels. PMID- 15123554 TI - Vulnerability to ventricular arrhythmias [corrected] and heterogeneity of action potential duration in normal rats. AB - In normal rats, we analysed the arrhythmogenic role of intrinsic action potential duration (APD) heterogeneity. In each animal, ventricular arrhythmic events (VAEs) occurring spontaneously and during the exposure to an acute social challenge were telemetrically recorded. Action potentials were recorded from isolated left ventricular myocytes, at a pacing rate of 5 Hz (patch clamp: current-clamp mode). APDs were measured at -20 mV, -30 mV, -40 mV, -50 mV and -60 mV. The difference between the shortest and the longest APD was also computed, as an index of individual APD heterogeneity. Animals predisposed to stress-induced arrhythmias showed higher values of APD and APD heterogeneity as compared with the remaining rats. We concluded that, in the normal heart, a large intrinsic APD heterogeneity resulting from specific electrophysiological properties of ventricular myocytes is not in itself arrhythmogenic, but can predispose towards arrhythmia development under certain conditions, such as autonomic activation. PMID- 15123555 TI - Re-innervation and recovery of rat soleus muscle and motor unit function after nerve crush. AB - In this study we have investigated the effects of peripheral nerve crush on the contractile properties of the adult rat soleus muscle. The soleus nerve was crushed close to the muscle and functional re-innervation was assessed by the measurement of the force produced by contraction induced by electrical nerve stimulation. Whole soleus muscle and single motor unit (MU) properties were studied at increasing re-innervation times 7-56 days after crush. Results showed progressive re-innervation as 50% of the axotomized motoneurones had re innervated their muscle 7 days after crush, 72% at 14 days and re-innervation was complete at 28 days. The force parameters recovered more slowly. Tetanic contractions faded at high frequency stimulation, which did not occur in the control muscle. This disruption in the tetanic response was more pronounced in single MUs. Our results demonstrate for the first time a process of progressive axonal re-innervation by the axotomized motoneurones and provide a functional picture of the effective restoration of the neuromuscular function. PMID- 15123556 TI - The effects of insulin on mesenteric blood flow in anaesthetized pigs. AB - Infusion of insulin in anaesthetized pigs has been shown to cause an increase in renal blood flow and a decrease in coronary blood flow, which were the net result of a vasoconstriction involving sympathetic alpha-adrenoceptor-mediated mechanisms and of a local vasodilatation involving the endothelial release of nitric oxide. In the present study, the effect of insulin on superior mesenteric blood flow was examined in pentobarbitone-anaesthetized pigs at constant heart rate, aortic blood pressure, left ventricular contractility and blood levels of glucose and potassium. In 10 pigs, infusion of 0.004 IU kg(-1) min(-1) of insulin increased mesenteric flow. In five of these pigs, intravenous phentolamine enhanced the increase in mesenteric flow elicited by insulin, a response which was abolished by the subsequent injection of N(omega)-nitro-L-arginine methyl ester (L-NAME) into the mesenteric artery. In the remaining five pigs, infusion of insulin after intramesenteric injection of L-NAME caused a decrease in mesenteric flow. This response was abolished by the subsequent intravenous administration of phentolamine. The present study showed that infusion of insulin in anaesthetized pigs primarily caused a mesenteric vasodilatation, which was the net result of two opposite effects, namely a predominant vasodilatation mediated by the endothelial release of nitric oxide and a sympathetic vasoconstrictor mechanism mediated by alpha-adrenoceptors. PMID- 15123557 TI - The expanding family of -RFamide peptides and their effects on feeding behaviour. AB - Neuropeptides terminating in -Arg-Phe-NH(2) (-RFamide) were first discovered in molluscan nervous systems, but were soon recognized to occur widely throughout the invertebrates. Progress in characterizing members of the family in vertebrates has been slower. In mammals, however, it is now clear that there are at least five genes encoding members of the family, and at least five G-protein coupled receptors at which they act. The tissue distribution of the peptides and their receptors is wide and there are likely to be many different functions. One of the emerging themes from recent research is that these peptides are involved in control of feeding behaviour both in invertebrates and in vertebrates. This would seem to be a remarkable example of conservation of chemical structure and biological function throughout nervous system evolution. PMID- 15123558 TI - Differential expression of the mechanosensitive potassium channel TREK-1 in epicardial and endocardial myocytes in rat ventricle. AB - Mechanoelectric feedback (MEF) is the process by which mechanical forces on the myocardium induce electrical responses. It is thought that MEF is important in controlling the beat to beat force of contraction in the ventricle, in response to fluctuations in load, and it may also play a role in controlling the dispersion of repolarization. The transduction mechanism for MEF is via stretch sensitive ion channels in the surface membrane of myocytes. Two types of stretch sensitive channels have been described; a non-selective cation channel, and a potassium selective channel. TREK-1 is a member of the recently cloned tandem pore potassium channels that has been shown to be mechanosensitive and to be expressed in rat heart. Here we report that the gene expression level of TREK-1, quantified using real-time RT-PCR against glyceraldehyde phosphate dehydrogenase (GAPDH) as a comparator gene, was found to be 0.34 +/- 0.14 in endocardial cells compared to 0.02 +/- 0.02 in epicardial cells (P < 0.05). To confirm that this is reflected in a different current density, whole cell TREK-1 currents, activated by chloroform, were recorded with patch clamp techniques in epicardial and endocardial cells. TREK-1 current density in epicardial and endocardial cells was 0.21 +/- 0.06 pA/pF and 0.8 +/- 0.27 pA/pF, respectively (P 0.05)]. It was concluded that the faster pulmonary O(2) kinetics during heavy KE exercise following prior heavy exercise was not associated with a similar modulation in the BF to the working muscles. PMID- 15123560 TI - Interactions between nitric oxide and superoxide on the neural regulation of proximal fluid reabsorption in hypertensive rats. AB - This study investigated the role of nitric oxide (NO) and superoxide anions in modulating the renal nerve-dependent increases in proximal tubular fluid reabsorption (Jva). Renal nerve stimulation at 0.75 and 1.0 Hz (15 V, 0.2 ms) in anaesthetized Wistar rats had no effect on glomerular filtration rate but decreased urine flow and sodium excretion in a frequency-related manner, reaching 39 and 49% at 1.0 Hz, respectively (P < 0.01) and increased Jva by 11 and 31% (P < 0.01). In the stroke prone spontaneously hypertensive rats (SHRSP), basal mean blood pressure was higher (123 +/- 2 versus 99 +/- 2 mmHg, P < 0.001), glomerular filtration rate, urine flow, sodium excretion and proximal tubular fluid reabsorption (Jva) were lower (all P < 0.001) than in the Wistar rats. Renal nerve stimulation in the SHRSP did not change glomerular filtration rate but decreased urine flow, and sodium excretion by 18 and 34% (P < 0.05) at 1.0 Hz which was less (P < 0.05) than that in the Wistar rats. Under these conditions, Jva was increased at 0.75 Hz by 27%, and to a comparable extent at 1.0 Hz, which was a pattern very different from the frequency related rises reported in the Wistar rats. In the SHRSP, intratubular Nomega-nitro-L-arginine methyl ester (L NAME) had no effect on baseline Jva or the pattern of response to renal nerve stimulation which contrasted with earlier reports in the Wistar rat. Intraluminal superoxide dismutase (SOD) had no effect on basal Jva in the Wistar rats but increased it in the SHRSP (P < 0.05) while the pattern of change in Jva during nerve stimulation was unaltered in both rat strains. By contrast, in the SHRSP, intraluminal sodium nitroprusside (SNP) resulted in a frequency related increase in Jva comparable to that obtained in the vehicle treated Wistar rats. These data suggest that in the hypertensive rats, superoxide anion production is raised which depresses Jva and interacts with NO preventing a normal Jva response to renal nerve stimulation. PMID- 15123561 TI - Evidence of lactoferrin transportation into blood circulation from intestine via lymphatic pathway in adult rats. AB - Using adult rats, the characteristic transporting system for lactoferrin (LF) from intestinal lumen into the blood circulation was investigated. The rats were randomly divided into two groups, a non-collected thoracic lymph (NC) group and a collected thoracic lymph (LC) group. Peripheral blood and thoracic lymph were collected from a jugular vein and a thoracic lymph duct, respectively, under anaesthesia. Bovine LF (bLF) was infused into the duodenal lumen by needle over a 1-min period at a dose of 1 g kg(-1). The transported bLF in the plasma and lymph was assayed quantitatively by double-antibody enzyme-linked immunosorbent assay (ELISA). Morphological investigation was also carried out in the intestine, lymph node, and liver. Following intraduodenal administration of bLF, the transported bLF in the NC group was detected in the plasma, and reached a peak value at 2 h. Furthermore, the bLF concentration in the thoracic duct lymph fluid in the LC group increased significantly, and peaked 2 h after the administration. In addition, bLF was not detected in the plasma of the LC group. Immunohistochemical analysis clearly showed anti-bLF positive particles in the epithelial cells of the apical villi. The striated border and baso-lateral membrane were also bLF positive. These results suggest that intraduodenally infused bLF is transported into the blood circulation via the lymphatic pathway, not via portal circulation in adult rats. PMID- 15123562 TI - The CSF and arterial to internal jugular venous hormonal differences during exercise in humans. AB - Strenuous exercise increases the cerebral uptake of carbohydrate out of proportion to that of oxygen, but it is unknown whether such enhanced carbohydrate uptake is influenced by the marked endocrine response to exercise. During exhaustive exercise this study evaluated the a-v differences across the brain (a-v diff) of hormones that could influence its carbohydrate uptake (n= 9). In addition, neuroendocrine activity and a potential uptake of hormones via the cerebrospinal fluid (CSF) were assessed by lumbar puncture postexercise and at rest (n= 6). Exercise increased the arterial concentration of noradrenaline and adrenaline, but there was no cerebral uptake. However, following exercise CSF noradrenaline was 1.4 (0.73-5.5) nmol l(-1), and higher than at rest, 0.3 (0.19 1.84) nmol l(-1) (P < 0.05), whereas adrenaline could not be detected. Exercise increased both the arterial concentration of NH(4)(+) and its a-v diff, which increased from 1 (-12 to 5) to 17 (5-41) micromol l(-1) (P < 0.05), while the CSF NH(4)(+) was reduced to 7 (0-10) versus 11 (7-16) micromol l(-1) (P < 0.05). There was no release from, or accumulation in the brain of interleukin (IL)-6, tumour necrosis factor (TNF-alpha), heatshock protein (HSP72), insulin, or insulin-like growth factor (IGF)-I. The findings indicate that for maximal exercise, the concentration of noradrenaline is increased within the brain, whereas blood borne hormones and cytokines are seemingly unimportant. The results support the notion that the exercise-induced changes in brain metabolism are controlled by factors intrinsic to the brain. PMID- 15123563 TI - NMDA receptor antagonism blocks the cardiovascular responses to microinjection of trans-ACPD into the NTS of awake rats. AB - The possible interaction of glutamatergic metabotropic agonists and N-methyl-d aspartate (NMDA) receptors was investigated in the nucleus tractus solitarii (NTS) of awake rats. The cardiovascular responses to unilateral microinjection of trans-1-amino-1,3-cyclopentanediocarboxylic acid (trans-ACPD; 250 pmol/50 nL) into the NTS (n= 8) produced hypotension (-64 +/- 4 mmHg) and bradycardic (-206 +/- 11 bpm) responses, which were blocked by previous microinjection of 2-amino-5 phosphonovaleric acid (AP-5; 10 nmol/50 nL), a selective antagonist of NMDA ionotropic receptors, into the same site. Intravenous injection of methyl atropine blocked both the bradycardic and hypotensive responses to microinjection of trans-ACPD into the NTS, indicating that the hypotension was secondary to the intense bradycardic response. The data also showed that the bradycardic and hypotensive responses to microinjection of an NMDA agonist (10 pmol/50 nL) into the NTS were not affected by previous microinjection of alpha-methyl-4 carboxyphenylglycine (MCPG; 5 nmol/50 nL), a non-selective antagonist of metabotropic receptors. The results showing that the cardiovascular responses to microinjection of trans-ACPD into the NTS were blocked by AP-5 indicate that the responses to metabotropic agonists in the NTS involves NMDA receptors. PMID- 15123564 TI - The effect of changes in arterial PCO2 on neuroendocrine function in man. AB - There is evidence that changes in arterial P(CO(2)) (P(a,CO(2))), as well as P(O(2)), influence neuroendocrine function. The hyponatraemia and fluid retention (cor pumonale) seen in chronic obstructive pulmonary disease (COPD) and type II respiratory failure is associated with increased vasopressin release. This study examines the specific effects of altered P(a,CO(2)) on hormone release from the posterior and anterior pituitary. The study was performed in 20 ventilated ICU patients in the late recovery phase of their illness. None had primary respiratory disease. Control blood samples were taken and the alveolar ventilation was then adjusted to allow the P(a,CO(2)) increase or decrease for a period of 3 h, during which time further blood samples were taken for the determination, by radioimmmunoassy of vasopressin, oxytocin, growth hormone and cortisol. Urine output and electrolyte concentrations were also measured. Circulating concentrations of growth hormone and oxytocin increased with increasing P(a,CO(2)). Vasopressin release showed a similar pattern up to a P(a,CO(2)) of approximately 6.0 kPa, above which vasopressin concentrations were inversely related to P(a,CO(2)). There was no significant effect on cortisol concentrations. No significant effects were established in urinary parameters during the short period of this study. Thus an increase in CO(2) is associated with stimulated pituitary hormone release. The effect on the neurohypophysial hormones may account for the fluid retention and hyponatraemia seen in COPD and hence provide a rationale for treatment. PMID- 15123565 TI - The effect of hypoxia on pulmonary O2 uptake, leg blood flow and muscle deoxygenation during single-leg knee-extension exercise. AB - The effect of hypoxic breathing on pulmonary O(2) uptake (VO(2p)), leg blood flow (LBF) and O(2) delivery and deoxygenation of the vastus lateralis muscle was examined during constant-load single-leg knee-extension exercise. Seven subjects (24 +/- 4 years; mean +/-s.d.) performed two transitions from unloaded to moderate-intensity exercise (21 W) under normoxic and hypoxic (P(ET)O(2)= 60 mmHg) conditions. Breath-by-breath VO(2p) and beat-by-beat femoral artery mean blood velocity (MBV) were measured by mass spectrometer and volume turbine and Doppler ultrasound (VingMed, CFM 750), respectively. Deoxy-(HHb), oxy-, and total haemoglobin/myoglobin were measured continuously by near-infrared spectroscopy (NIRS; Hamamatsu NIRO-300). VO(2p) data were filtered and averaged to 5 s bins at 20, 40, 60, 120, 180 and 300 s. MBV data were filtered and averaged to 2 s bins (1 contraction cycle). LBF was calculated for each contraction cycle and averaged to 5 s bins at 20, 40, 60, 120, 180 and 300 s. VO(2p) was significantly lower in hypoxia throughout the period of 20, 40, 60 and 120 s of the exercise on transient. LBF (l min(-1)) was approximately 35% higher (P > 0.05) in hypoxia during the on-transient and steady-state of KE exercise, resulting in a similar leg O(2) delivery in hypoxia and normoxia. Local muscle deoxygenation (HHb) was similar in hypoxia and normoxia. These results suggest that factors other than O(2) delivery, possibly the diffusion of O(2,) were responsible for the lower O(2) uptake during the exercise on-transient in hypoxia. PMID- 15123566 TI - Nitric oxide and thyroid gland: modulation of cardiovascular function in autonomic-blocked anaesthetized rats. AB - We have previously reported that acute administration of N(G)-nitro-l-arginine methyl ester (L-NAME) increases the mean arterial pressure (MAP) and heart rate (HR) in autonomic-blocked (CAB) anaesthetized rats. In the present study we examined whether thyroid and adrenal glands are involved in these pressor and chronotropic responses. Sprague-Dawley rats were studied after bilateral vagotomy and ganglionic blockade with hexamethonium (10 mg kg(-1)), and stabilization of MAP with infusion of phenylephrine (PE) (6 microg kg(-1) min(-1)). The rats were divided into groups: L, CAB; PE, CAB + PE bolus (6 microg kg(-1)); L-TX, thyroidectomy + CAB; L-AX, adrenalectomy + CAB; TX, only thyroidectomy; C, CAB. L, L-AX and L-TX groups received a bolus of l-NAME (7.5 mg kg(-1)). Triiodothyronine (T3), thyroxin (T4) and thyrotropin (TSH) levels were measured in L and L-TX rats before and after l-NAME administration. Reduced nicotamide adenine dinucleotide (NADPH) diaphorase activity was determined in heart and aorta of the TX group. The pressor response induced by l-NAME was similar in all groups. l-NAME-induced-tachycardia was associated with this rise in MAP. Adrenalectomy did not modify this chronotropic response, but it was attenuated by thyroidectomy. Thyroidectomy by itself decreased the circulating levels of T3 but it had no effect on the plasma levels of T4 and TSH. L and L-TX groups showed similar levels of circulating T4 and TSH, meanwhile the plasma level of T3 decreased in the L group. Nitric oxide synthase (NOS) activity in atria as well as in aorta was greater in the TX group compared with C. When autonomic influences are removed, the thyroid gland modulates intrinsic heart rate via a mechanism that involves, at least in part, the nitric oxide pathway. PMID- 15123567 TI - Elevated blood pressure in normotensive rats produced by 'knockdown' of the angiotensin type 2 receptor. AB - Most of our knowledge of the function of the angiotensin type 2 receptor (AT(2)R) has been obtained from transgenic mouse models. The aim of the present study was to investigate the role of the AT(2)R in normotensive Sprague-Dawley (SD) rats by using antisense gene transfer technology to 'knockdown' this specific receptor subtype. A retroviral vector containing full-length AT(2)R antisense cDNA (AT(2)R AS) was constructed and the effectiveness of the transduction of AT(2)R-AS was studied in vitro. In subsequent in vivo studies, 5-day-old normotensive SD rats received a single intracardiac bolus (25 microl) of AT(2)R-AS viral particles. When animals reached adulthood, direct blood pressure (BP), and both pressor and dipsogenic responses to angiotensin II were investigated. Long-lasting expression of the AT(2)R-AS transcript and a reduction in mRNA and binding of the AT(2)R was observed in vitro. Expression of AT(2)R-AS transcript was maintained for 90 days in heart, kidney, lung and brain, indicating a high degree of transgene transduction in vivo. As adults, systolic BP and the pressor responses to angiotensin were significantly elevated in AT(2)R-AS-treated rats. However, AT(2)R-AS-treated rats displayed significantly reduced dipsogenic responses to both angiotensin and water deprivation. Collectively, these data demonstrate that a single neonatal injection of the retroviral vector containing antisense to the AT(2) receptors in rats results in similar cardiovascular and dipsogenic responses as reported in AT(2)R knockout mice. The actions of the AT(2) receptors appear to be antagonistic to the cardiovascular actions of the AT(1) receptors, whereas AT(1) and AT(2) receptors appear to act synergistically in the regulation of water intake. PMID- 15123568 TI - Role of angiotensin II in the pressor response to cortisol in fetal sheep during late gestation. AB - Glucocorticoids increase blood pressure in utero, but the mechanisms responsible are unclear. This study investigated the hypothesis that the hypertensive effects of cortisol depend upon a functional renin-angiotensin system (RAS). The study examined, in the sheep fetus, whether blockade of the Ang II type 1 (AT(1)) specific receptor prevented the cortisol-induced increase in blood pressure. From 124 +/- 1 days of gestation (term 145 +/- 2 days), 27 chronically catheterized sheep fetuses were infused i.v. for 5 days with one of the following: (1) saline (0.9% NaCl at 2.5 ml day(-1), n= 6); (2) cortisol (3-5 mg kg(-1) day(-1), n= 7); (3) AT(1) receptor antagonist (GR138950, 1-3 mg kg(-1) day(-1) in saline, GRS, n= 7); or (4) cortisol and GR138950 (GRC, n= 7). On all days of infusion, plasma cortisol was greater in both groups of cortisol-treated fetuses than in the respective control fetuses (P < 0.05), and GR138950 prevented the pressor response to exogenous Ang II. Over 5 days of infusion, blood pressure increased by a maximum of 7.6 +/- 1.4 mmHg (mean +/-s.e.m., P < 0.05) in the cortisol-, but not saline-infused, fetuses. Blockade of the AT(1) receptor caused significant reductions in blood pressure in both GRS- and GRC-treated groups (P < 0.05); in the GRS-treated fetuses, the fall in blood pressure was significant from the first day of infusion, while in GRC-treated fetuses the decrement was not significant until the second day (P < 0.05). Over the period of the infusion, decreases in arterial blood pH andP(a,O(2)), and an increase inP(a,CO(2)), were observed in the fetuses treated with the AT(1) receptor antagonist (P < 0.05). Therefore, in the sheep fetus, 5 days of AT(1) receptor antagonism suppresses the cortisol-induced rise in blood pressure. These results suggest that cortisol may increase blood pressure within 24 h of administration by a mechanism that is independent of the fetal RAS. Thereafter, Ang II, via the AT(1) receptor, may mediate, in part, the hypertensive effects of cortisol in utero. PMID- 15123569 TI - Left ventricular hypertrophy is more common in black than white hypertensives: is this news? PMID- 15123570 TI - Hypoadiponectinemia is an independent risk factor for hypertension. AB - Adiponectin is one of the key molecules in the metabolic syndrome, and its concentration is decreased in obesity, type-2 diabetes, and coronary artery disease. Genetic investigation has revealed that 2 polymorphisms (I164T and G276T) are related to adiponectin concentration and diabetes. To examine whether adiponectin affects hypertension genetically or biologically, we performed a case control study. A total of 446 diagnosed cases of hypertension (HT) in men and 312 normotensive (NT) men were enrolled in this study. Plasma adiponectin concentration was measured using an enzyme-linked immunosorbent assay system. Single nucleotide polymorphisms were determined by TaqMan polymerase chain reaction method. After adjustment for confounding factors, adiponectin concentration was significantly lower in HT (HT: 5.2+/-0.2 microg/mL; NT: 6.1+/ 0.2 microg/mL; P<0.001). Furthermore, multiple regression analysis indicated that hypoadiponectinemia was an independent risk factor for hypertension (P<0.001). Blood pressure was inversely associated with adiponectin concentration in normotensives regardless of insulin resistance. In subjects carrying the TC genotype of the I164T polymorphism, adiponectin concentration was significantly lower (TC: 2.6+/-0.9 microg/mL; TT: 5.5+/-0.1 microg/mL; P<0.01), and most of them had hypertension. In contrast, the G276T polymorphism was not associated with adiponectin concentration or hypertension. In conclusion, hypoadiponectinemia is a marker for predisposition to hypertension in men. PMID- 15123571 TI - Insulin resistance and hypertension: the Insulin Resistance Atherosclerosis study. AB - The association between insulin resistance and insulinemia and hypertension is controversial. We examined the relation between insulin resistance and hypertension in 564 non-Hispanic whites (NHW), 505 Hispanics (H), and 413 African Americans (AA) who participated in the Insulin Resistance Atherosclerosis Study (IRAS). Insulin sensitivity was measured with a frequently sampled intravenous glucose tolerance test with minimal model analysis. The prevalence of hypertension was 32.5%, 49.4%, and 32.3% in NHW, AA, and H, respectively (P<0.001). When subjects without diabetes in all ethnic groups were combined, age, male sex, race (AA), body mass index (BMI), and insulin resistance, but not fasting insulin, were significantly associated with hypertension. When each ethnic group was analyzed separately, insulin resistance was significantly associated with hypertension in NHW and H, but not AA. After excluding subjects taking antihypertensive medications, male sex, BMI, fasting glucose, and insulin resistance, but not fasting insulin, were significant determinants of blood pressure. When the 3 ethnic groups were analyzed separately, insulin resistance was significantly associated with blood pressure in H, but not NHW, or AA. Neither insulin resistance nor fasting insulin was significantly associated with hypertension or blood pressure in subjects with diabetes of the 3 ethnic groups after adjusting for age, sex, BMI, and waist. In conclusion, insulin resistance, but not insulinemia, was related to hypertension and blood pressure in subjects without diabetes, but ethnic differences in these relations appear to exist. Neither insulin resistance nor insulinemia was related to hypertension or blood pressure in patients with type 2 diabetes in the 3 ethnic groups. PMID- 15123572 TI - Changes in arterial stiffness and wave reflection with advancing age in healthy men and women: the Framingham Heart Study. AB - With advancing age, arterial stiffness and wave reflections increase and elevate systolic and pulse pressures. An elevated central pulse pressure is generally ascribed to increased wave reflection and portends an unfavorable prognosis. Using arterial tonometry, we evaluated central (carotid-femoral) and peripheral (carotid-brachial) pulse wave velocity, amplitudes of forward and reflected pressure waves, and augmentation index in 188 men and 333 women in the Framingham Heart Study offspring cohort who were free of clinical cardiovascular disease, hypertension, diabetes, smoking within the past 12 months, dyslipidemia, and obesity. In multivariable linear regression models, advancing age was the predominant correlate of higher carotid-femoral pulse wave velocity; other correlates were higher mean arterial pressure, heart rate, and triglycerides and walk test before tonometry (model R2=0.512, P<0.001). A similar model was obtained for carotid-brachial pulse wave velocity (model R2=0.227, P<0.001), although the increase with advancing age was smaller. Owing to different relations of age to central and peripheral stiffness measures, carotid-femoral pulse wave velocity was lower than carotid-brachial pulse wave velocity before age 50 years but exceeded it thereafter, leading to reversal of the normal central-to-peripheral arterial stiffness gradient. In this healthy cohort with a minimal burden of cardiovascular disease risk factors, an age-related increase in aortic stiffness, as compared with peripheral arterial stiffness, was associated with increasing forward wave amplitude and pulse pressure and reversal of the arterial stiffness gradient. This phenomenon may facilitate forward transmission of potentially deleterious pressure pulsations into the periphery. PMID- 15123573 TI - Differences in left ventricular structure between black and white hypertensive adults: the Hypertension Genetic Epidemiology Network study. AB - The degree to which ethnic differences in left ventricular structure among hypertensive adults are independent of clinical and hemodynamic factors remains uncertain. We assessed whether left ventricular mass and geometry differ between black and white hypertensives after accounting for differences in such factors. Our study group comprised 1060 black and 580 white hypertensive participants free of valvular or coronary disease in a population-based cohort. Blood pressure was measured during a clinic visit and echocardiography was performed using standardized protocols. After controlling for clinical and hemodynamic parameters (cardiac index, peripheral resistance index, and pulse pressure/stroke index), both left ventricular mass and relative wall thickness were higher in blacks than whites (173.9+/-30.9 versus 168.3+/-24.3 grams, P=0.006, and 0.355+/-0.055 versus 0.340+/-0.055 grams, P<0.001). Similarly, the adjusted risk of having left ventricular hypertrophy, whether indexed by height(2.7) or by body surface area, was greater for blacks than for whites (odds ratio: 1.80; 95% CI: 1.29 to 2.51; and odds ratio: 2.50; 95% CI: 1.58 to 3.96, respectively), and this was also true for concentric geometry (odds ratio: 2.28; 95% CI: 1.22 to 4.25). Further adjustment for relatedness in this genetic epidemiological study did not attenuate these differences. Our findings confirm the strong association between black ethnicity and increased left ventricular mass and relative wall thickness in hypertensive adults and demonstrate that these differences are independent of standard clinical and hemodynamic parameters. Whether such differences relate to distinct ambulatory pressure profiles or an ethnic propensity to cardiac hypertrophy requires further investigation. PMID- 15123574 TI - C3 or not C3: that is the question. PMID- 15123575 TI - Stimulation of cyclic GMP production via AT2 and B2 receptors in the pressure overloaded aorta after banding. AB - Abdominal aortic banding induces upregulation of the angiotensin II (Ang II) type 2 (AT2) receptor, thereby decreasing the contractile response to Ang II in the thoracic aorta of the rat. The aim of this study was to use a mouse model to clarify the mechanisms by which the banding elicits upregulation of the aortic AT2 receptor and the subsequent attenuation of Ang II responsiveness. Concomitantly with the elevation in blood pressure and plasma renin concentration after banding, AT2-receptor mRNA levels in the thoracic aorta rapidly increased in mice within 4 days. Upregulation of the AT2 receptor, as well as blood pressure elevation after banding, was abolished by losartan administration. The contractile response to Ang II was depressed in aortic rings of banding mice but not of sham mice, and was restored by either the AT2-receptor antagonist PD123319 or the bradykinin B2-receptor antagonist icatibant. cGMP content in the thoracic aorta of banding mice was 9-fold greater than that of sham mice, and the elevation was reduced to sham levels 1 hour after intravenous injection of PD123319 or icatibant. When aortic rings were incubated with Ang II, cGMP content increased in banding rings but not in sham rings; the pretreatment with PD123319 or icatibant inhibited Ang II-induced cGMP production. These results suggest that aortic banding induces upregulation of the AT2 receptor through increased circulating Ang II via the AT1 receptor, thereby activating a vasodilatory pathway in vessels through the AT2 receptor via the kinin/cGMP system. PMID- 15123576 TI - Role of hypothalamic melanocortin 3/4-receptors in mediating chronic cardiovascular, renal, and metabolic actions of leptin. AB - The present study examined whether blockade of melanocortin receptors subtypes 3 and 4 (MC3/4-R) inhibits chronic cardiovascular and dietary responses to leptin infusion. A cannula was placed in the lateral ventricle of male Sprague-Dawley rats for chronic intracerebroventricular (ICV) infusion via osmotic minipump, and arterial and venous catheters were implanted for measurement of mean arterial pressure (MAP) and heart rate (HR) 24 h/d and IV infusions. After a 5-day control period, rats received (1) 0.9% saline vehicle ICV for 12 days plus leptin (1 microg/kg per minute IV, n=5) during the final 7 days; (2) MC3/4-R antagonist SHU 9119 (1 nmol/h ICV) for 12 days plus leptin (1 microg/kg per minute IV, n=6) during the final 7 days; and (3) SHU-9119 (1 nmol/h ICV, n=8) for 12 days. Leptin infusion in vehicle-treated rats caused a small increase in MAP (5+/-1 mm Hg) despite reduced food intake (23+/-1 to 10+/-1 g/d) and decreased body weight ( 6%+/-1%). SHU-9119 infusion completely prevented the cardiovascular and dietary actions of leptin, leading to increased food intake (23+/-1 to 49+/-4 g/d) and body weight (+30%+/-2%), markedly decreased HR (-77+/-9 bpm), and caused a decrease in MAP (-6+/-1 mm Hg). Similar results were observed when SHU-9119 was infused alone in vehicle-treated rats. Leptin decreased plasma insulin to 30% of control values, an effect that was also abolished by SHU-9119 treatment, which caused a 5-fold increase in plasma insulin concentration. Thus, MC3/4-R antagonism completely blocked the chronic cardiovascular, satiety, and metabolic effects of leptin, suggesting that the hypothalamic melanocortin system plays an important role in mediating these actions of leptin. PMID- 15123577 TI - Left ventricle mass index and the common, functional, X-linked angiotensin II type-2 receptor gene polymorphism (-1332 G/A) in patients with systemic hypertension. AB - A common intronic polymorphism, (-1332 G/A) of the angiotensin type-2 receptor gene, located on the X-chromosome, has been reported to be functional. The aim of our study was to evaluate this polymorphism for an association with left ventricular hypertrophy. Left ventricle (LV) mass was measured in 197 patients with systemic hypertension and 60 normal volunteers using a 1.5-Tesla Philips MRI system. Genotyping was performed using a restriction enzyme digestion of an initial 310-bp polymerase chain reaction product that included the angiotensin type-2 (-1332 G/A) locus. The mean LV mass index for the male patients was 94.3+/ 19.6 g/m2 (n=125) and for the female patients was 71.2+/-12.0 g/m2 (n=72). Seventy-three (37.1%) of all patients had an elevated LV mass index, defined as the mean LV mass index for normal volunteers plus 2 SD (males 77.8+/-9.1 g/m2, n=30; females 61.5+/-7.5 g/m2, n=30). Comparison of LV mass index of the A_/AA genotype (mean LV mass index=82.4+/-21.1 g/m2; n=123) against that of the G_/GG genotype (mean LV mass index=88.1+/-19.0 g/m2; n=89) as a continuous variable was significant by ANOVA (P=0.044). chi2 Comparison between subjects with and subjects without left ventricular hypertrophy revealed an excess of the G_/GG genotype among the group with LV hypertrophy (P=0.031). We observed an association between the angiotensin type-2 receptor (-1332 G) allele and the presence of left ventricular hypertrophy in hypertensive subjects. PMID- 15123578 TI - Role of osteopontin in cardiac fibrosis and remodeling in angiotensin II-induced cardiac hypertrophy. AB - Osteopontin (OPN) is upregulated in several experimental models of cardiac fibrosis and remodeling. However, its direct effects remain unclear. We examined the hypothesis that OPN is important for the development of cardiac fibrosis and remodeling. Moreover, we examined whether the inhibitory effect of eplerenone (Ep), a novel aldosterone receptor antagonist, was mediated through the inhibition of OPN expression against cardiac fibrosis and remodeling. Wild-type (WT) and OPN-deficient mice were treated with angiotensin II (Ang II) for 4 weeks. WT mice receiving Ang II were divided into 2 groups: a control group and an Ep treatment group. Ang II treatment significantly elevated blood pressure and caused cardiac hypertrophy and fibrosis in WT mice. Ep treatment and OPN deficiency could reduce the Ang II-induced elevation of blood pressure and ameliorate the development of cardiac fibrosis, whereas Ep-only treatment abolished the development of cardiac hypertrophy. Most compelling, the reduction of cardiac fibrosis led to an impairment of cardiac systolic function and subsequent left ventricular dilatation in Ang II-treated OPN-deficient mice. These results suggest that OPN has a pivotal role in the development of Ang II induced cardiac fibrosis and remodeling. Moreover, the effect of Ep on the prevention of cardiac fibrosis, but not cardiac hypertrophy, might be partially mediated through the inhibition of OPN expression. PMID- 15123579 TI - Analysis of segmental duplications and genome assembly in the mouse. AB - Limited comparative studies suggest that the human genome is particularly enriched for recent segmental duplications. The extent of segmental duplications in other mammalian genomes is unknown and confounded by methodological differences in genome assembly. Here, we present a detailed analysis of recent duplication content within the mouse genome using a whole-genome assembly comparison method and a novel assembly independent method, designed to take advantage of the reduced allelic variation of the C57BL/6J strain. We conservatively estimate that approximately 57% of all highly identical segmental duplications (>or=90%) were misassembled or collapsed within the working draft WGS assembly. The WGS approach often leaves duplications fragmented and unassigned to a chromosome when compared with the clone-ordered-based approach. Our preliminary analysis suggests that 1.7%-2.0% of the mouse genome is part of recent large segmental duplications (about half of what is observed for the human genome). We have constructed a mouse segmental duplication database to aid in the characterization of these regions and their integration into the final mouse genome assembly. This work suggests significant biological differences in the architecture of recent segmental duplications between human and mouse. In addition, our unique method provides the means for improving whole-genome shotgun sequence assembly of mouse and future mammalian genomes. PMID- 15123580 TI - The functional genomic distribution of protein divergence in two animal phyla: coevolution, genomic conflict, and constraint. AB - We compare the functional spectrum of protein evolution in two separate animal lineages with respect to two hypotheses: (1) rates of divergence are distributed similarly among functional classes within both lineages, indicating that selective pressure on the proteome is largely independent of organismic-level biological requirements; and (2) rates of divergence are distributed differently among functional classes within each lineage, indicating species-specific selective regimes impact genome-wide substitutional patterns. Integrating comparative genome sequence with data from tissue-specific expressed-sequence-tag (EST) libraries and detailed database annotations, we find a functional genomic signature of rapid evolution and selective constraint shared between mammalian and nematode lineages despite their extensive morphological and ecological differences and distant common ancestry. In both phyla, we find evidence of accelerated evolution among components of molecular systems involved in coevolutionary change. In mammals, lineage-specific fast evolving genes include those involved in reproduction, immunity, and possibly, maternal-fetal conflict. Likelihood ratio tests provide evidence for positive selection in these rapidly evolving functional categories in mammals. In contrast, slowly evolving genes, in terms of amino acid or insertion/deletion (indel) change, in both phyla are involved in core molecular processes such as transcription, translation, and protein transport. Thus, strong purifying selection appears to act on the same core cellular processes in both mammalian and nematode lineages, whereas positive and/or relaxed selection acts on different biological processes in each lineage. PMID- 15123582 TI - A global view of the selection forces in the evolution of yeast cis-regulation. AB - The interaction between transcription factors and their DNA binding sites is key to understanding gene regulation. By performing a genome-wide study of the evolutionary dynamics in yeast promoters, we provide a first global view of the network of selection forces in the evolution of transcription factor binding sites. This analysis gives rise to new models for binding site activity, identifies families of related binding sites, and characterizes the functional similarities among them. We discovered rich and highly optimized selective pressures operating inside and around these families. In several cases, this organization reveals that a single transcription factor has multiple functional modes. We demonstrate how such functional heterogeneity is related to the binding site's affinity and how it is exploited in transcription programs. PMID- 15123581 TI - The Human MitoChip: a high-throughput sequencing microarray for mitochondrial mutation detection. AB - Somatic mitochondrial mutations are common in human cancers, and can be used as a tool for early detection of cancer. We have developed a mitochondrial Custom Reseq microarray as an array-based sequencing platform for rapid and high throughput analysis of mitochondrial DNA. The MitoChip contains oligonucleotide probes synthesized using standard photolithography and solid-phase synthesis, and is able to sequence >29 kb of double-stranded DNA in a single assay. Both strands of the entire human mitochondrial coding sequence (15,451 bp) are arrayed on the MitoChip; both strands of an additional 12,935 bp (84% of coding DNA) are arrayed in duplicate. We used 300 ng of genomic DNA to amplify the mitochondrial coding sequence in three overlapping long PCR fragments. We then sequenced >2 million base pairs of mitochondrial DNA, and successfully assigned base calls at 96.0% of nucleotide positions. Replicate experiments demonstrated >99.99% reproducibility. In matched fluid samples (urine and pancreatic juice, respectively) obtained from five patients with bladder cancer and four with pancreatic cancer, the MitoChip detected at least one cancer-associated mitochondrial mutation in six (66%) of nine samples. The MitoChip is a high-throughput sequencing tool for the reliable identification of mitochondrial DNA mutations from primary tumors in clinical samples. PMID- 15123583 TI - Dynamics of a human interparalog gene conversion hotspot. AB - Gene conversion between paralogs can alter their patterns of sequence identity, thus obscuring their evolutionary relationships and affecting their propensity to sponsor genomic rearrangements. The details of this important process are poorly understood in the human genome because allelic diversity complicates the interpretation of interparalog sequence differences. Here we exploit the haploid nature of the Y chromosome, which obviates complicating interallelic processes, together with its known phylogeny, to understand the dynamics of conversion between two directly repeated HERVs flanking the 780-kb AZFa region on Yq. Sequence analysis of a 787-bp segment of each of the HERVs in 36 Y chromosomes revealed one of the highest nucleotide diversities in the human genome, as well as evidence of a complex patchwork of highly directional gene conversion events. The rate of proximal-to-distal conversion events was estimated as 2.4 x 10(-4) to 1.2 x 10(-3) per generation (3.9 x 10(-7) to 1.9 x 10(-6) per base per generation), and the distal-to-proximal rate as about one-twentieth of this. Minimum observed conversion tract lengths ranged from 1 to 158 bp and maximum lengths from 19 to 1365 bp, with an estimated mean of 31 bp. Analysis of great ape homologs shows that conversion in this hotspot has a deep evolutionary history. PMID- 15123584 TI - Testing the chromosomal speciation hypothesis for humans and chimpanzees. AB - Fixed differences of chromosomal rearrangements between isolated populations may promote speciation by preventing between-population gene flow upon secondary contact, either because hybrids suffer from lowered fitness or, more likely, because recombination is reduced in rearranged chromosomal regions. This chromosomal speciation hypothesis thus predicts more rapid genetic divergence on rearranged than on colinear chromosomes because the former are less porous to gene flow. A number of studies of fungi, plants, and animals, including limited genetic data of humans and chimpanzees, support the hypothesis. Here we reexamine the hypothesis for humans and chimpanzees with substantially more genomic data than were used previously. No difference is observed between rearranged and colinear chromosomes in the level of genomic DNA sequence divergence between species. The same is also true for protein sequences. When the gorilla is used as an outgroup, no acceleration in protein sequence evolution associated with chromosomal rearrangements is found. Furthermore, divergence in expression pattern between orthologous genes is not significantly different for rearranged and colinear chromosomes. These results, showing that chromosomal rearrangements did not affect the rate of genetic divergence between humans and chimpanzees, are expected if incipient species on the evolutionary lineages separating humans and chimpanzees did not hybridize. PMID- 15123585 TI - A versatile assay for high-throughput gene expression profiling on universal array matrices. AB - We report a flexible, sensitive, and quantitative gene-expression profiling system for assaying more than 400 genes, with three probes per gene, for 96 samples in parallel. The cDNA-mediated annealing, selection, extension and ligation (DASL) assay targets specific transcripts, using oligonucleotides containing unique address sequences that can hybridize to universal arrays. Cell specific gene expression profiles were obtained using this assay for hormone treated cell lines and laser-capture microdissected cancer tissues. Gene expression profiles derived from this assay were consistent with those determined by qRT-PCR. The DASL assay has been automated for use with a bead-based 96-array matrix system. The combined high-throughput assay and readout system is accurate and efficient, and can cost-effectively profile the expression of hundreds of genes in thousands of samples. PMID- 15123586 TI - Compositional gene landscapes in vertebrates. AB - The existence of a well conserved linear relationship between GC levels of genes' second and third codon positions (GC2, GC3) prompted us to focus on the landscape, or joint distribution, spanned by these two variables. In human, well curated coding sequences now cover at least 15%-30% of the estimated total gene set. Our analysis of the landscape defined by this gene set revealed not only the well documented linear crest, but also the presence of several peaks and valleys along that crest, a property that was also indicated in two other warm-blooded vertebrates represented by large gene databases, that is, mouse and chicken. GC2 is the sum of eight amino acid frequencies, whereas GC3 is linearly related to the GC level of the chromosomal region containing the gene. The landscapes therefore portray relations between proteins and the DNA environments of the genes that encode them. PMID- 15123587 TI - Two methods of whole-genome amplification enable accurate genotyping across a 2320-SNP linkage panel. AB - Comprehensive genome scans involving many thousands of SNP assays will require significant amounts of genomic DNA from each sample. We report two successful methods for amplifying whole-genomic DNA prior to SNP analysis, multiple displacement amplification, and OmniPlex technology. We determined the coverage of amplification by analyzing a SNP linkage marker set that contained 2320 SNP markers spread across the genome at an average distance of 2.5 cM. We observed a concordance of >99.8% in genotyping results from genomic DNA and amplified DNA, strongly indicating the ability of both methods used to amplify genomic DNA in a highly representative manner. Furthermore, we were able to achieve a SNP call rate of >98% in both genomic and amplified DNA. The combination of whole-genome amplification and comprehensive SNP linkage analysis offers new opportunities for genetic analysis in clinical trials, disease association studies, and archiving of DNA samples. PMID- 15123588 TI - The Ensembl core software libraries. AB - Systems for managing genomic data must store a vast quantity of information. Ensembl stores these data in several MySQL databases. The core software libraries provide a practical and effective means for programmers to access these data. By encapsulating the underlying database structure, the libraries present end users with a simple, abstract interface to a complex data model. Programs that use the libraries rather than SQL to access the data are unaffected by most schema changes. The architecture of the core software libraries, the schema, and the factors influencing their design are described. All code and data are freely available. PMID- 15123589 TI - The Ensembl analysis pipeline. AB - The Ensembl pipeline is an extension to the Ensembl system which allows automated annotation of genomic sequence. The software comprises two parts. First, there is a set of Perl modules ("Runnables" and "RunnableDBs") which are 'wrappers' for a variety of commonly used analysis tools. These retrieve sequence data from a relational database, run the analysis, and write the results back to the database. They inherit from a common interface, which simplifies the writing of new wrapper modules. On top of this sits a job submission system (the "RuleManager") which allows efficient and reliable submission of large numbers of jobs to a compute farm. Here we describe the fundamental software components of the pipeline, and we also highlight some features of the Sanger installation which were necessary to enable the pipeline to scale to whole-genome analysis. PMID- 15123590 TI - The Ensembl automatic gene annotation system. AB - As more genomes are sequenced, there is an increasing need for automated first pass annotation which allows timely access to important genomic information. The Ensembl gene-building system enables fast automated annotation of eukaryotic genomes. It annotates genes based on evidence derived from known protein, cDNA, and EST sequences. The gene-building system rests on top of the core Ensembl (MySQL) database schema and Perl Application Programming Interface (API), and the data generated are accessible through the Ensembl genome browser (http://www.ensembl.org). To date, the Ensembl predicted gene sets are available for the A. gambiae, C. briggsae, zebrafish, mouse, rat, and human genomes and have been heavily relied upon in the publication of the human, mouse, rat, and A. gambiae genome sequence analysis. Here we describe in detail the gene-building system and the algorithms involved. All code and data are freely available from http://www.ensembl.org. PMID- 15123591 TI - The Ensembl Web site: mechanics of a genome browser. AB - The Ensembl Web site (http://www.ensembl.org/) is the principal user interface to the data of the Ensembl project, and currently serves >500,000 pages (approximately 2.5 million hits) per week, providing access to >80 GB (gigabyte) of data to users in more than 80 countries. Built atop an open-source platform comprising Apache/mod_perl and the MySQL relational database management system, it is modular, extensible, and freely available. It is being actively reused and extended in several different projects, and has been downloaded and installed in companies and academic institutions worldwide. Here, we describe some of the technical features of the site, with particular reference to its dynamic configuration that enables it to handle disparate data from multiple species. PMID- 15123592 TI - Sockeye: a 3D environment for comparative genomics. AB - Comparative genomics techniques are used in bioinformatics analyses to identify the structural and functional properties of DNA sequences. As the amount of available sequence data steadily increases, the ability to perform large-scale comparative analyses has become increasingly relevant. In addition, the growing complexity of genomic feature annotation means that new approaches to genomic visualization need to be explored. We have developed a Java-based application called Sockeye that uses three-dimensional (3D) graphics technology to facilitate the visualization of annotation and conservation across multiple sequences. This software uses the Ensembl database project to import sequence and annotation information from several eukaryotic species. A user can additionally import their own custom sequence and annotation data. Individual annotation objects are displayed in Sockeye by using custom 3D models. Ensembl-derived and imported sequences can be analyzed by using a suite of multiple and pair-wise alignment algorithms. The results of these comparative analyses are also displayed in the 3D environment of Sockeye. By using the Java3D API to visualize genomic data in a 3D environment, we are able to compactly display cross-sequence comparisons. This provides the user with a novel platform for visualizing and comparing genomic feature organization. PMID- 15123593 TI - The otter annotation system. AB - With the completion of the human genome sequence and genome sequence available for other vertebrate genomes, the task of manual annotation at the large genome scale has become a priority. Possibly even more important, is the requirement to curate and improve this annotation in the light of future data. For this to be possible, there is a need for tools to access and manage the annotation. Ensembl provides an excellent means for storing gene structures, genome features, and sequence, but it does not support the extra textual data necessary for manual annotation. We have extended Ensembl to create the Otter manual annotation system. This comprises a relational database schema for storing the manual annotation data, an application-programming interface (API) to access it, an extensible markup language (XML) format to allow transfer of the data, and a server to allow multiuser/multimachine access to the data. We have also written a data-adaptor plugin for the Apollo Browser/Editor to enable it to utilize an Otter server. The otter database is currently used by the Vertebrate Genome Annotation (VEGA) site (http://vega.sanger.ac.uk), which provides access to manually curated human chromosomes. Support is also being developed for using the AceDB annotation editor, FMap, via a perl wrapper called Lace. The Human and Vertebrate Annotation (HAVANA) group annotators at the Sanger center are using this to annotate human chromosomes 1 and 20. PMID- 15123594 TI - The Ensembl computing architecture. AB - Ensembl is a software project to automatically annotate large eukaryotic genomes and release them freely into the public domain. The project currently automatically annotates 10 complete genomes. This makes very large demands on compute resources, due to the vast number of sequence comparisons that need to be executed. To circumvent the financial outlay often associated with classical supercomputing environments, farms of multiple, lower-cost machines have now become the norm and have been deployed successfully with this project. The architecture and design of farms containing hundreds of compute nodes is complex and nontrivial to implement. This study will define and explain some of the essential elements to consider when designing such systems. Server architecture and network infrastructure are discussed with a particular emphasis on solutions that worked and those that did not (often with fairly spectacular consequences). The aim of the study is to give the reader, who may be implementing a large-scale biocompute project, an insight into some of the pitfalls that may be waiting ahead. PMID- 15123595 TI - ESTGenes: alternative splicing from ESTs in Ensembl. AB - We describe a novel algorithm for deriving the minimal set of nonredundant transcripts compatible with the splicing structure of a set of ESTs mapped on a genome. Sets of ESTs with compatible splicing are represented by a special type of graph. We describe the algorithms for building the graphs and for deriving the minimal set of transcripts from the graphs that are compatible with the evidence. These algorithms are part of the Ensembl automatic gene annotation system, and its results, using ESTs, are provided at www.ensembl.org as ESTgenes for the mosquito, Caenorhabditis briggsae, C. elegans, zebrafish, human, mouse, and rat genomes. Here we also report on the results of this method applied to the human and mouse genomes. PMID- 15123596 TI - GeneWise and Genomewise. AB - We present two algorithms in this paper: GeneWise, which predicts gene structure using similar protein sequences, and Genomewise, which provides a gene structure final parse across cDNA- and EST-defined spliced structure. Both algorithms are heavily used by the Ensembl annotation system. The GeneWise algorithm was developed from a principled combination of hidden Markov models (HMMs). Both algorithms are highly accurate and can provide both accurate and complete gene structures when used with the correct evidence. PMID- 15123597 TI - Munc13-1-mediated vesicle priming contributes to secretory amyloid precursor protein processing. AB - The amyloid precursor protein (APP) gives rise toc beta-amyloid peptides, which are the main constituents of senile plaques in brains of Alzheimer's disease patients. Non-amyloidogenic processing of the APP can be stimulated by phorbol esters (PEs) and by intracellular diacylglycerol (DAG) generation. This led to the hypothesis that classical and novel protein kinase Cs (PKCs), which are activated by DAG/PEs, regulate APP processing. However, in addition to PKCs, there are other DAG/PE receptors present in neurons that may participate in the modulation of APP processing. Munc13-1, a presynaptic protein with an essential role in synaptic vesicle priming, represents such an alternative target of the DAG second messenger pathway. Using Munc13-1 knock-out mice and knock-in mice expressing a Munc13-1(H567K) variant deficient in DAG/PE binding, we determined the relative contributions of PKCs and Munc13-1 to PE-stimulated secretory APP processing. We establish that, in addition to PKC, Munc13-1 significantly contributes to the regulation of secretory APP metabolism. PMID- 15123598 TI - The presenilin proteins are components of multiple membrane-bound complexes that have different biological activities. AB - Several lines of evidence have indicated that the presenilin proteins function within macromolecular complexes and are necessary for the regulated intramembranous proteolysis of certain type 1 transmembrane proteins, including the amyloid precursor protein, Notch, and p75. Data from multiple complementary experiments now suggest that there may be several distinct presenilin complexes. We show here that presenilin mutations and certain detergents affect the abundance and componentry of the presenilin complexes, and these structural effects correlate with their effects on gamma-secretase activity. Our data suggest that there are at least three complexes, including a approximately 150 kDa nicastrin-aph-1 complex (which is likely to be a precursor complex). There is a stable and abundant intermediate complex of approximately 440 kDa, which contains aph-1, pen-2, nicastrin, and PS1. However, it is the very low abundance, high mass (>/=670 kDa) heteromeric complexes that are associated with the highest gamma-secretase-specific activity. PMID- 15123599 TI - Synaptotagmins I and II act as nerve cell receptors for botulinum neurotoxin G. AB - Botulinum neurotoxins (BoNTs) induce muscle paralysis by selectively entering cholinergic motoneurons and subsequent specific cleavage of core components of the vesicular fusion machinery. Complex gangliosides are requisite for efficient binding to neuronal cells, but protein receptors are critical for internalization. Recent work evidenced that synaptotagmins I and II can function as protein receptors for BoNT/B (Dong, M., Richards, D. A., Goodnough, M. C., Tepp, W. H., Johnson, E. A., and Chapman, E. R. (2003) J. Cell Biol. 162, 1293 1303). Here, we report the protein receptor for a second BoNT serotype. Like BoNT/B, BoNT/G employs synaptotagmins I and II to enter phrenic nerve cells. Using pull-down assays we show that only BoNT/G, but neither the five remaining BoNTs nor tetanus neurotoxin, interacts with synaptotagmins I and II. In contrast to BoNT/B, interactions with both isoforms are independent of the presence of gangliosides. Peptides derived from the luminal domain of synaptotagmin I and II are capable of blocking the neurotoxicity of BoNT/G in phrenic nerve preparations. Pull-down and neutralization assays further established the membrane-juxtaposed 10 luminal amino acids of synaptotagmins I and II as the critical segment for neurotoxin binding. In addition, we show that the carboxyl terminal domain of the cell binding fragment of BoNT/B and BoNT/G mediates the interaction with their protein receptor. PMID- 15123600 TI - Identification of the mitochondrial ATP-Mg/Pi transporter. Bacterial expression, reconstitution, functional characterization, and tissue distribution. AB - The mitochondrial carriers are a family of transport proteins that, with a few exceptions, are found in the inner membranes of mitochondria. They shuttle metabolites, nucleotides, and cofactors through this membrane and thereby connect and/or regulate cytoplasm and matrix functions. ATP-Mg is transported in exchange for phosphate, but no protein has ever been associated with this activity. We have isolated three human cDNAs that encode proteins of 458, 468, and 489 amino acids with 66-75% similarity and with the characteristic features of the mitochondrial carrier family in their C-terminal domains and three EF-hand Ca(2+) binding motifs in their N-terminal domains. These proteins have been overexpressed in Escherichia coli and reconstituted into phospholipid vesicles. Their transport properties and their targeting to mitochondria demonstrate that they are isoforms of the ATP-Mg/Pi carrier described in the past in whole mitochondria. The tissue specificity of the three isoforms shows that at least one isoform was present in all of the tissues investigated. Because phosphate recycles via the phosphate carrier in mitochondria, the three isoforms of the ATP Mg/Pi carrier are most likely responsible for the net uptake or efflux of adenine nucleotides into or from the mitochondria and hence for the variation in the matrix adenine nucleotide content, which has been found to change in many physiopathological situations. PMID- 15123601 TI - An Hsp27-related, dominant-negative-acting intracellular estradiol-binding protein. AB - New World primates (NWPs) exhibit a compensated form of resistance to gonadal steroid hormones. We demonstrated recently that estrogen resistance in NWP cells was associated with the overexpression of two proteins, a nonreceptor-related, dominant-negative-acting estrogen response element (ERE)-binding protein (ERE-BP) and an intracellular estradiol-binding protein (IEBP). Based on the N-terminal sequences of tryptic fragments of IEBP isolated from a 17beta-estradiol (E2) affinity column we cloned a full-length cDNA for IEBP from the estrogen-resistant NWP cell line, B95-8. Subsequent sequence analysis revealed 87% sequence identity between the deduced peptide for IEBP and human Hsp27. When hormone-responsive, wild-type Old World primate (OWP) cells were transiently transfected with IEBP cDNA, E2-directed ERE reporter luciferase activity was reduced by 50% compared with vector only-transfected OWP cells (p < 0.0018). When IEBP and ERE-BP were cotransfected, ERE promoter-reporter activity was reduced by a further 60% (p < 0.0001). Electrophoresis mobility shift analyses showed that IEBP neither bound to ERE nor competed with the estrogen receptor (ER) for binding to ERE. However, there was evidence of protein-protein interaction of IEBP and ERalpha; IEBP was coimmunoprecipitated with anti-ERalpha antibody in wild-type cells stably transfected with IEBP. A specific interaction between ERalpha and IEBP was confirmed in glutathione S-transferase pull-down and yeast two-hybrid assays. Data indicate that the Hsp27-related IEBP interacts with the ligand binding domain of the ERalpha. In summary, by inhibiting the ERalpha-E2 interaction, IEBP acts to squelch ERalpha-directed ERE-regulated transactivation and promote estrogen resistance in NWP cells. PMID- 15123602 TI - Solution structure of the tandem Src homology 3 domains of p47phox in an autoinhibited form. AB - The phagocyte NADPH oxidase is a multisubunit enzyme responsible for the generation of superoxide anions (O(2).) that kill invading microorganisms. p47(phox) is a cytosolic subunit of the phagocyte NADPH oxidase, which plays a crucial role in the assembly of the activated NADPH oxidase complex. The molecular shapes of the p47(phox) tandem SH3 domains either with or without a polybasic/autoinhibitory region (PBR/AIR) at the C terminus were studied using small angle x-ray scattering. The tandem SH3 domains with PBR/AIR formed a compact globular structure, whereas the tandem SH3 domains lacking the PBR/AIR formed an elongated structure. Alignment anisotropy analysis by NMR based on the residual dipolar couplings revealed that the tandem SH3 domains with PBR/AIR were in good agreement with a globular module corresponding to the split half of the intertwisted dimer in crystalline state. The structure of the globular module was elucidated to represent a solution structure of the tandem SH3 domain in the autoinhibited form, where the PBR/AIR bundled the tandem SH3 domains and the linker forming a closed structure. Once PBR/AIR is released by phosphorylation, rearrangements of the SH3 domains may occur, forming an open structure that binds to the cytoplasmic proline-rich region of membrane-bound p22(phox). PMID- 15123603 TI - Direct photoaffinity labeling by dolastatin 10 of the amino-terminal peptide of beta-tubulin containing cysteine 12. AB - Tubulin with bound [5-3H]dolastatin 10 was exposed to ultraviolet light, and 8 10% of the bound drug cross-linked to the protein, most of it specifically. The primary cross-link was to the peptide spanning amino acid residues 2-31 of beta tubulin, but the specific amino acid could not be identified. Indirect studies indicated that cross-link formation occurred between cysteine 12 and the thiazole moiety of dolastatin 10. An equipotent analog of dolastatin 10, lacking the thiazole ring, did not form an ultraviolet light-induced cross-link to beta tubulin. Preillumination of tubulin with ultraviolet light, known to induce cross link formation between cysteine 12 and exchangeable site nucleotide, inhibited the binding of [5-3H]dolastatin 10 and cross-link formation more potently than it inhibited the binding of colchicine or vinblastine to tubulin. Conversely, binding of dolastatin 10 to tubulin inhibited formation of the cross-link between cysteine 12 and the exchangeable site nucleotide. Dithiothreitol inhibited formation of the beta-tubulin/dolastatin 10 cross-link but not the beta tubulin/exchangeable site nucleotide cross-link. Modeling studies revealed a highly favored binding site for dolastatin 10 at the + end of beta-tubulin in proximity to the exchangeable site GDP. Computational docking of an energy minimized dolastatin 10 conformation at this site placed the thiazole ring of dolastatin 10 8-9 A from the sulfur atom of cysteine 12. Dolastatin 15 and cryptophycin 1 could also be docked into positions that overlapped more extensively with the docked dolastatin 10 than with each other. This result was consistent with the observed binding properties of these peptides. PMID- 15123604 TI - A M55V polymorphism in a novel SUMO gene (SUMO-4) differentially activates heat shock transcription factors and is associated with susceptibility to type I diabetes mellitus. AB - Three SUMO (small ubiquitin-related modifier) genes have been identified in humans, which tag proteins to modulate subcellular localization and/or enhance protein stability and activity. We report the identification of a novel intronless SUMO gene, SUMO-4, that encodes a 95-amino acid protein having an 86% amino acid homology with SUMO-2. In contrast to SUMO-2, which is highly expressed in all of the tissues examined, SUMO-4 mRNA was detected mainly in the kidney. A single nucleotide polymorphism was detected in SUMO-4, substituting a highly conserved methionine with a valine residue (M55V). In HepG2 (liver carcinoma) cells transiently transfected with SUMO-4 expression vectors, Met-55 was associated with the elevated levels of activated heat shock factor transcription factors as compared with Val-55, whereas the levels of NF-kappaB were suppressed to an identical degree. The SUMO-4M (Met) variant is associated with type I diabetes mellitus susceptibility in families (p = 4.0 x 10(-4)), suggesting that it may be involved in the pathogenesis of type I diabetes. PMID- 15123605 TI - Insulin/Foxo1 pathway regulates expression levels of adiponectin receptors and adiponectin sensitivity. AB - Adiponectin/Acrp30 is a hormone secreted by adipocytes, which acts as an antidiabetic and antiatherogenic adipokine. We reported previously that AdipoR1 and -R2 serve as receptors for adiponectin and mediate increased fatty acid oxidation and glucose uptake by adiponectin. In the present study, we examined the expression levels and roles of AdipoR1/R2 in several physiological and pathophysiological states such as fasting/refeeding, obesity, and insulin resistance. Here we show that the expression of AdipoR1/R2 in insulin target organs, such as skeletal muscle and liver, is significantly increased in fasted mice and decreased in refed mice. Insulin deficiency induced by streptozotocin increased and insulin replenishment reduced the expression of AdipoR1/R2 in vivo. Thus, the expression of AdipoR1/R2 appears to be inversely correlated with plasma insulin levels in vivo. Interestingly, the incubation of hepatocytes or myocytes with insulin reduced the expression of AdipoR1/R2 via the phosphoinositide 3 kinase/Foxo1-dependent pathway in vitro. Moreover, the expressions of AdipoR1/R2 in ob/ob mice were significantly decreased in skeletal muscle and adipose tissue, which was correlated with decreased adiponectin binding to membrane fractions of skeletal muscle and decreased AMP kinase activation by adiponectin. This adiponectin resistance in turn may play a role in worsening insulin resistance in ob/ob mice. In conclusion, the expression of AdipoR1/R2 appears to be inversely regulated by insulin in physiological and pathophysiological states such as fasting/refeeding, insulin deficiency, and hyper-insulinemia models via the insulin/phosphoinositide 3-kinase/Foxo1 pathway and is correlated with adiponectin sensitivity. PMID- 15123606 TI - The hepatitis B virus X protein inhibits secretion of apolipoprotein B by enhancing the expression of N-acetylglucosaminyltransferase III. AB - The X protein of hepatitis B virus (HBx) plays a major role on hepatocellular carcinoma (HCC). Apolipoprotein B (apoB) in the liver is an important glycoprotein for transportation of very low density lipoproteins and low density lipoproteins. Although lipid accumulation in the liver is known as one of the factors for the HCC, the relationship between HBx and apoB during the HCC development is poorly understood. To better understand the biological significance of HBx in HCC, liver Chang cells that specifically express HBx were established and characterized. In this study we demonstrate that overexpression of HBx significantly up-regulates the expression of UDP-N-acetylglucosamine:beta d-mannoside-1,4-N-acetylglucosaminyltransferase-III (GnT-III), an enzyme that functions as a bisecting-N-acetylglucosamine (GlcNAc) transferase in apoB, and increases GnT-III promoter activity in a chloramphenicol acetyltransferase assay. GnT-III expression levels of HBx-transfected cells appeared to be higher than that of hepatocarcinoma cells as well as GnT-III-transfected cells, indicating that HBx may has a strong GnT-III promotor-enhancing activity. Intracellular levels of apoBs, which contained the increased bisecting GlcNAc, were accumulated in HBx-transfected liver cells. These cells as well as GnT-III-transfected liver cells revealed the inhibition of apoB secretion and the increased accumulation of intracellular triglyceride and cholesterol compared with vector-transfected cells. Moreover, overexpression of GnT-III and HBx in liver cells was shown to down-regulate the transcriptional level of microsomal triglyceride transfer protein, which regulates the assembly and secretion of apoB. Therefore, our study strongly suggested that the HBx increase in intracellular accumulation of aberrantly glycosylated apoB resulted in inhibition of secretion of apoB as well as intracellular lipid accumulation by elevating the expression of GnT-III. PMID- 15123607 TI - Transposon mutagenesis of Trypanosoma brucei identifies glycosylation mutants resistant to concanavalin A. AB - We have engineered Trypanosoma brucei with a novel mariner transposition system that allows large populations of mutant cells to be generated and screened. As a proof of principle, we isolated and characterized two independent clones that were resistant to the cytotoxic action of concanavalin A. In both clones, the transposon had integrated into the locus encoding a homologue of human ALG12, which encodes a dolichyl-P-Man: Man(7)GlcNAc(2)-PP-dolichyl-alpha6 mannosyltransferase. Conventional knock-out of ALG12 in a wild-type background gave an identical phenotype to the mariner mutants, and biochemical analysis confirmed that they have the same defect in the N-linked oligosaccharide synthesis pathway. To our surprise, both mariner mutants were homozygous; the second allele appeared to have undergone gene conversion by the mariner-targeted allele. Subsequent experiments showed that the frequency of gene conversion at the ALG12 locus, in the absence of selection, was 0.25%. As we approach the completion of the trypanosome genome project, transposon mutagenesis provides an important addition to the repertoire of genetic tools for T. brucei. PMID- 15123608 TI - Lipin expression preceding peroxisome proliferator-activated receptor-gamma is critical for adipogenesis in vivo and in vitro. AB - We recently identified mutations in the lipin gene, Lpin1, as the cause of lipodystrophy in the fatty liver dystrophy (fld) mouse. Here we identify impaired adipocyte differentiation as the basis for lipodystrophy in lipin-deficient mice and demonstrate that lipin is required for normal induction of the adipogenic gene transcription program. We found that the reduced adiposity in chow fed fld mice and resistance to obesity in fld mice fed a high-fat diet is associated with reduced adipogenic gene expression. Using primary mouse embryonic fibroblasts isolated from fld mice, we confirmed that lipin deficiency prevents normal lipid accumulation and induction of key adipogenic genes, including peroxisome proliferator-activated receptor (PPAR)gamma and CCAAT enhancer-binding protein (C/EBP)alpha. However, our previous studies of daily gene expression in differentiating 3T3-L1 preadipocytes indicated that lipin expression is undetectable until about day 3 of differentiation, at a point after PPARgamma and C/EBPalpha gene expression is established. This paradox was resolved by examining gene expression at 10-h intervals during 3T3-L1 cell differentiation, leading to detection of transient lipin expression at 10 h into the differentiation program, prior to the induction of PPARgamma and C/EBPalpha. Consistent with a requirement for lipin expression upstream of PPARgamma, differentiation of lipin-deficient mouse embryonic fibroblasts could be rescued by ectopic expression of PPARgamma. Thus, we conclude that lipin expression is required prior to PPARgamma during adipocyte differentiation. PMID- 15123609 TI - A conserved DpYR motif in the juxtamembrane domain of the Met receptor family forms an atypical c-Cbl/Cbl-b tyrosine kinase binding domain binding site required for suppression of oncogenic activation. AB - The activation and phosphorylation of Met, the receptor tyrosine kinase (RTK) for hepatocyte growth factor, initiates the recruitment of multiple signaling proteins, one of which is c-Cbl, a ubiquitin-protein ligase. c-Cbl promotes ubiquitination and enhances the down-modulation of the Met receptor and other RTKs, targeting them for lysosomal sorting and subsequent degradation. The ubiquitination of Met by c-Cbl requires the direct interaction of the c-Cbl tyrosine kinase binding (TKB) domain with tyrosine 1003 in the Met juxtamembrane domain. Although a consensus for c-Cbl TKB domain binding has been established ((D/N)XpYXX(D/E0phi), this motif is not present in Met, suggesting that other c Cbl TKB domain binding motifs may exist. By alanine-scanning mutagenesis, we have identified a DpYR motif including Tyr(1003) as being important for the direct recruitment of the c-Cbl TKB domain and for ubiquitination of the Met receptor. The substitution of Tyr(1003) with phenylalanine or substitution of either aspartate or arginine residues with alanine impairs c-Cbl-recruitment and ubiquitination of Met and results in the oncogenic activation of the Met receptor. We demonstrate that the TKB domain of Cbl-b, but not Cbl-3, binds to the Met receptor and requires an intact DpYR motif. Modeling studies suggest the presence of a salt bridge between the aspartate and arginine residues that would position pTyr(1003) for binding to the c-Cbl TKB domain. The DpYR motif is conserved in other members of the Met RTK family but is not present in previously identified c-Cbl-binding proteins, identifying DpYR as a new binding motif for c Cbl and Cbl-b. PMID- 15123610 TI - Identification of two binding regions for the suppressor of hairless protein within the intracellular domain of Drosophila notch. AB - Notch is a phylogenetically conserved transmembrane receptor that is required for many aspects of animal development. Upon ligand stimulation, a fragment of Notch is released proteolytically and enters the nucleus to form a complex with the DNA binding protein CSL (CBF1/Suppressor of Hairless/Lag1) and activate transcription of Notch-CSL target genes. The physical structure of the Notch-CSL complex remains unclear, however, clouding the interpretation of previous efforts to correlate Notch structure and function. We have, therefore, characterized the binding of Drosophila CSL (called Suppressor of Hairless, or Su(H)) to the intracellular domain of Drosophila Notch both in vitro and in vivo. We report the identification of two Su(H) binding regions in Notch. The first is in the juxtamembrane region (the "RAM" domain). The second is just C-terminal to the Notch ankyrin repeats, overlapping or identical to two previously proposed nuclear localization sequences, in a domain we term PPD (potential phosphorylated domain). The ankyrin repeats themselves do not bind to Su(H); however, they substantially enhance binding of Su(H) to the more C-terminal region. Consistent with this picture, removal of either the Ram or PPD binding sites, separately, modestly reduces Notch activity in vivo, whereas removal of both renders Notch severely defective. These results clarify the relationship between Notch and CSL, help to explain the importance of the ankyrin repeats in Notch signaling, and reconcile many apparently contradictory results from previous Notch structure/function studies. Moreover, they suggest a second function for the Notch nuclear localization sequence elements. PMID- 15123611 TI - Actions downstream of cyclic GMP/protein kinase G can reverse protein kinase C mediated phosphorylation of CPI-17 and Ca2+ sensitization in smooth muscle. AB - Ca(2+) sensitivity of smooth muscle contraction is modulated by several systems converging on myosin light chain phosphatase (MLCP). Rho-Rho kinase is considered to inhibit MLCP via phosphorylation, whereas protein kinase C (PKC) induced sensitization has been shown to be dependent on phosphorylation of the inhibitory protein CPI-17. We have explored the interaction of cGMP-dependent protein kinase (PKG) with Ca(2+) sensitization pathways using permeabilized mouse smooth muscle. Three conditions giving approximately 50% of maximal active force were compared in small intestinal preparations: 1). Ca(2+)-activated unsensitized muscle (pCa 5.9 with Rho kinase inhibitor Y27632); 2). Rho-Rho kinase-sensitized muscle (pCa 6.1 with guanosine 5'-3-O-(thio)triphosphate); and 3). PKC-sensitized muscle (pCa 6.0 with Y27632 and PKC activator phorbol 12,13-dibutyrate). 8-Br-cGMP relaxed the sensitized muscles but had marginal effects on unsensitized preparations, showing that PKG reverses both PKC and Rho-mediated Ca(2+) sensitization. CPI-17 was present in permeabilized intestinal tissue. In PKC-sensitized preparations, CPI-17 phosphorylation decreased in response to 8-Br-cGMP. The rate of PKC mediated phosphorylation in the presence of the MLCP inhibitor microcystin-LR was not influenced by 8-Br-cGMP. PKC-induced Ca(2+) sensitization also was reversed in vascular smooth muscle tissues (portal vein and femoral artery). We conclude that actions downstream of cGMP/PKG can reverse PKC-mediated phosphorylation of CPI-17 and Ca(2+) sensitization in smooth muscle. PMID- 15123612 TI - Direct probing of copper active site and free radical formed during bicarbonate dependent peroxidase activity of bovine and human copper, zinc-superoxide dismutases. Low-temperature electron paramagnetic resonance and electron nuclear double resonance studies. AB - Using X-band electron paramagnetic resonance (EPR) and electron nuclear double resonance (ENDOR) spectroscopy at liquid helium temperatures, the Cu(II) coordination geometry at the active site of bovine and human copper,zinc superoxide dismutases (bSOD1 and hSOD1) treated with H(2)O(2) and bicarbonate (HCO(3)(-)) was examined. The time course EPR of wild type human SOD1 (WT hSOD1), W32F hSOD1 mutant (tryptophan 32 substituted with phenylalanine), and bSOD1 treated with H(2)O(2) and HCO(3)(-) shows an initial reduction of active site Cu(II) to Cu(I) followed by its oxidation back to Cu(II) in the presence of H(2)O(2). However, HCO(3)(-) induced a Trp-32-derived radical from WT hSOD1 but not from bSOD1. The mutation of Trp-32 by phenylalanine totally eliminated the Trp-32 radical signal generated from W32F hSOD1 treated with HCO(3)(-) and H(2)O(2). Further characterization of the free radical was performed by UV irradiation of WT hSOD1 and bSOD1 that generated tryptophanyl and tyrosyl radicals. Both proton ((1)H) and nitrogen ((14)N) ENDOR studies of bSOD1 and hSOD1 in the presence of H(2)O(2) revealed a change in the geometry of His-46 (or His-44) and His-48 (or His-46) coordinated to Cu(II) at the active site of WT hSOD1 and bSOD1, respectively. However, in the presence of HCO(3)(-) and H(2)O(2), both (1)H and (14)N ENDOR spectra were almost identical to those derived from native bSOD1. We conclude that HCO(3)(-)-derived oxidant does not alter significantly the Cu(II) active site geometry and histidine coordination to Cu(II) in SOD1 as does H(2)O(2) alone; however, the oxidant derived from HCO(3)( ) (i.e. carbonate anion radical) reacts with surface-associated Trp-32 in hSOD1 to form the corresponding radical. PMID- 15123613 TI - Oleoylethanolamide stimulates lipolysis by activating the nuclear receptor peroxisome proliferator-activated receptor alpha (PPAR-alpha). AB - Amides of fatty acids with ethanolamine (FAE) are biologically active lipids that participate in a variety of biological functions, including the regulation of feeding. The polyunsaturated FAE anandamide (arachidonoylethanolamide) increases food intake by activating G protein-coupled cannabinoid receptors. On the other hand, the monounsaturated FAE oleoylethanolamide (OEA) reduces feeding and body weight gain by activating the nuclear receptor PPAR-alpha (peroxisome proliferator-activated receptor alpha). In the present report, we examined whether OEA can also influence energy utilization. OEA (1-20 microm) stimulated glycerol and fatty acid release from freshly dissociated rat adipocytes in a concentration-dependent and structurally selective manner. Under the same conditions, OEA had no effect on glucose uptake or oxidation. OEA enhanced fatty acid oxidation in skeletal muscle strips, dissociated hepatocytes, and primary cardiomyocyte cultures. Administration of OEA in vivo (5 mg kg(-1), intraperitoneally) produced lipolysis in both rats and wild-type mice, but not in mice in which PPAR-alpha had been deleted by homologous recombination (PPAR alpha(-/-)). Likewise, OEA was unable to enhance lipolysis in adipocytes or stimulate fatty acid oxidation in skeletal muscle strips isolated from PPAR-alpha mice. The synthetic PPAR-alpha agonist Wy-14643 produced similar effects, which also were dependent on the presence of PPAR-alpha. Subchronic treatment with OEA reduced body weight gain and triacylglycerol content in liver and adipose tissue of diet-induced obese rats and wild-type mice, but not in obese PPAR-alpha(-/-) mice. The results suggest that OEA stimulates fat utilization through activation of PPAR-alpha and that this effect may contribute to its anti-obesity actions. PMID- 15123614 TI - Loss of srf-3-encoded nucleotide sugar transporter activity in Caenorhabditis elegans alters surface antigenicity and prevents bacterial adherence. AB - During the establishment of a bacterial infection, the surface molecules of the host organism are of particular importance, since they mediate the first contact with the pathogen. In Caenorhabditis elegans, mutations in the srf-3 locus confer resistance to infection by Microbacterium nematophilum, and they also prevent biofilm formation by Yersinia pseudotuberculosis, a close relative of the bubonic plague agent Yersinia pestis. We cloned srf-3 and found that it encodes a multitransmembrane hydrophobic protein resembling nucleotide sugar transporters of the Golgi apparatus membrane. srf-3 is exclusively expressed in secretory cells, consistent with its proposed function in cuticle/surface modification. We demonstrate that SRF-3 can function as a nucleotide sugar transporter in heterologous in vitro and in vivo systems. UDP-galactose and UDP-N acetylglucosamine are substrates for SRF-3. We propose that the inability of Yersinia biofilms and M. nematophilum to adhere to the nematode cuticle is due to an altered glycoconjugate surface composition of the srf-3 mutant. PMID- 15123615 TI - Dynamin interacts with members of the sumoylation machinery. AB - Dynamin is a GTP-binding protein whose oligomerization-dependent assembly around the necks of lipid vesicles mediates their scission from parent membranes. Dynamin is thus directly involved in the regulation of endocytosis. Sumoylation is a post-translational protein modification whereby the ubiquitin-like modifier Sumo is covalently attached to lysine residues on target proteins by a process requiring the concerted action of an activating enzyme (ubiquitin-activating enzyme), a conjugating enzyme (ubiquitin carrier protein), and a ligating enzyme (ubiquitin-protein isopeptide ligase). Here, we show that dynamin interacts with Sumo-1, Ubc9, and PIAS-1, all of which are members of the sumoylation machinery. Ubc9 and PIAS-1 are known ubiquitin carrier protein and ubiquitin-protein isopeptide ligase enzymes, respectively, for the process of sumoylation. We have identified the coiled-coil GTPase effector domain (GED) of dynamin as the site on dynamin that interacts with Sumo-1, Ubc9, and PIAS-1. Although we saw no evidence of covalent Sumo-1 attachment to dynamin, Sumo-1 and Ubc9 are shown here to inhibit the lipid-dependent oligomerization of dynamin. Expression of Sumo-1 and Ubc9 in mammalian cells down-regulated the dynamin-mediated endocytosis of transferrin, whereas dynamin-independent fluid-phase uptake was not affected. Furthermore, using high resolution NMR spectroscopy, we have identified amino acid residues on Sumo-1 that directly interact with the GED of dynamin. The results suggest that the GED of dynamin may serve as a scaffold that concentrates the sumoylation machinery in the vicinity of potential acceptor proteins. PMID- 15123616 TI - Crystal structure of the Toll/interleukin-1 receptor domain of human IL-1RAPL. AB - The Toll/interleukin-1 receptor (TIR) domain is conserved in the intracellular regions of Toll-like receptors (TLRs) and interleukin-1 receptors (IL-1Rs) as well as in several cytoplasmic adapter molecules. This domain has crucial roles in signal transduction by these receptors for host immune response. Here we report the crystal structure at 2.3-A resolution of the TIR domain of human IL 1RAPL, the first structure of a TIR domain of the IL-1R superfamily. There are large structural differences between this TIR domain and that of TLR1 and TLR2. Helix alphaD in IL-1RAPL is almost perpendicular to its equivalent in TLR1 or TLR2. The BB loop contains a hydrogen bond unique to IL-1RAPL between Thr residues at the 8th and 10th positions. The structural and sequence diversity among these domains may be important for specificity in the signal transduction by these receptors. A dimer of the TIR domain of IL-1RAPL is observed in the crystal, although this domain is monomeric in solution. Residues in the dimer interface are mostly unique to IL-1RAPL, which is consistent with the distinct functional roles of this receptor. Our functional studies show IL-1RAPL can activate JNK but not the ERK or the p38 MAP kinases, whereas its close homolog, TIGIRR, cannot activate JNK. Deletion mutagenesis studies show that the activation of JNK by IL-1RAPL does not depend on the integrity of its TIR domain, suggesting a distinct mechanism of signaling through this receptor. PMID- 15123617 TI - The expression of the phosphotyrosine phosphatase DEP-1/PTPeta dictates the responsivity of glioma cells to somatostatin inhibition of cell proliferation. AB - Here we characterize the intracellular effectors of the antiproliferative activity of somatostatin in glioma cell lines and post-surgical specimens. The responsiveness to somatostatin correlated with the expression of the phosphotyrosine phosphatase DEP-1/PTPeta, identified in C6 and U87MG cells, in which somatostatin inhibited cell growth. The expression of a dominant negative mutant of DEP-1/PTPeta in C6 cells abolished somatostatin effects, confirming the involvement of this phosphotyrosine phosphatase in such effects. Somatostatin treatment increased the activity of DEP-1/PTPeta and inhibited ERK1/2 activation. Conversely, basic fibroblast growth factor-dependent MEK phosphorylation was not affected, suggesting a direct effect on ERK1/2. In vitro experiments showed that PTPeta was able to interact and dephosphorylate ERK1/2 activated by basic fibroblast growth factor. Furthermore, by transfecting PTPeta in the somatostatin unresponsive, DEP-1/PTPeta-deficient U373MG cells, the somatostatin-dependent control of cell proliferation was recovered. Finally we evaluated the requirement for DEP-1/PTPeta in somatostatin inhibition of cell proliferation in post surgical specimens derived from different grade human gliomas. Although all of the glioma analyzed expressed somatostatin receptor mRNA, DEP-1/PTPeta expression was limited to 8 of 22 of the tumors. Culturing seven gliomas, a correlation between the expression of DEP-1/PTPeta and the somatostatin antiproliferative effects was identified. In conclusion we propose that the expression and activation of DEP-1/PTPeta is required for somatostatin inhibition of glioma proliferation. PMID- 15123618 TI - Involvement of Cdk5/p25 in digoxin-triggered prostate cancer cell apoptosis. AB - Cardiac digitalis has been considered to be a treatment for breast cancer. Our previous study indicates that digoxin, one member in digitalis, decreases the proliferation of prostate cancer cells, but the mechanisms remain unclear. In the present study, Ca(2+) proved to be an important factor in digoxin-triggered prostate cancer cell death. Because cyclin-dependent kinase (Cdk)5 and p35 cleavage (p25 formation) have been reported to be targets of intracellular Ca(2+), and subsequently correlated to apoptosis, we not only demonstrated first that Cdk5, p35, and p25 proteins were all expressed in prostate cancer cells (including lymph node carcinoma of the prostate (LNCaP) and DU-145 cells), but also showed where p25 formation and Cdk5 kinase activity were affected by treatment with digoxin. The inhibitor of p35 cleavage (calpeptin) was used to reduce p25 formation, and the result suggested that p25 accumulation might be the major cause of digoxin-triggered LNCaP cell death. Butyrolactone-I and roscovitine, two Cdk5 kinase inhibitors, were also found to prevent digoxin triggered LNCaP cell death. In addition, treatment of siRNA-Cdk5 diminished digoxin-triggered cell death, as compared with the treatments of siRNA-Cdk1 or siRNA-Cdk2, which implies the specific involvement of Cdk5 in digoxin-triggered cell death. Caspase inhibitor set and terminal deoxynucleotidyltransferase mediated dUTP nick end labeling assay were used to demonstrate that digoxin triggered LNCaP cell apoptosis through Cdk5 activation. These results suggest that Cdk5/p35 and p25 are novel players in digoxin-triggered prostate cancer cell apoptosis and, therefore, become potential therapeutic targets. PMID- 15123619 TI - Identification, expression, and purification of a pyrethroid-hydrolyzing carboxylesterase from mouse liver microsomes. AB - Carboxylesterases are enzymes that catalyze the hydrolysis of a wide range of ester-containing endogenous and xenobiotic compounds. Although the use of pyrethroids is increasing, the specific enzymes involved in the hydrolysis of these insecticides have yet to be identified. A pyrethroid-hydrolyzing enzyme was partially purified from mouse liver microsomes using a fluorescent reporter similar in structure to cypermethrin (Shan, G., and Hammock, B. D. (2001) Anal. Biochem. 299, 54-62 and Wheelock, C. E., Wheelock, A. M., Zhang, R., Stok, J. E., Morisseau, C., Le Valley, S. E., Green, C. E., and Hammock, B. D. (2003) Anal. Biochem. 315, 208-222) and subsequently identified as a carboxylesterase (NCBI accession number BAC36707). The expressed sequence tag was then cloned, expressed in baculovirus, and purified to homogeneity. Kinetic constants for a large number of both type I and type II pyrethroid or pyrethroid-like substrates were determined. This esterase possesses similar kinetic constants for cypermethrin and its fluorescent-surrogate (k(cat) = 0.12 +/- 0.03 versus 0.11 +/- 0.01 s( 1)). Compared with their cis- counterparts, trans-permethrin and cypermethrin were hydrolyzed 22- and 4-fold faster, respectively. Of the four fenvalerate isomers the (2R)(alphaR)-isomer was hydrolyzed at least 1 order of magnitude faster than any other isomer. However, it is unlikely that this enzyme accounts for the total pyrethroid hydrolysis in the microsomes because both isoelectrofocusing and native PAGE indicate the presence of a second region of cypermethrin-metabolizing enzymes. A second carboxylesterase gene (NCBI accession number NM_133960), isolated during a cDNA mouse liver library screening, was also found to hydrolyze pyrethroids. Both these enzymes could be used as preliminary tools in establishing the relative toxicity of new pyrethroids. PMID- 15123620 TI - Mice with a targeted disruption of the AE2 Cl-/HCO3- exchanger are achlorhydric. AB - The AE2 Cl-/HCO3- exchanger is expressed in numerous cell types, including epithelial cells of the kidney, respiratory tract, and alimentary tract. In gastric epithelia, AE2 is particularly abundant in parietal cells, where it may be the predominant mechanism for HCO3- efflux and Cl- influx across the basolateral membrane that is needed for acid secretion. To investigate the hypothesis that AE2 is critical for parietal cell function and to assess its importance in other tissues, homozygous null mutant (AE2(-/-)) mice were prepared by targeted disruption of the AE2 (Slc4a2) gene. AE2(-/-) mice were emaciated, edentulous (toothless), and exhibited severe growth retardation, and most of them died around the time of weaning. AE2(-/-) mice exhibited achlorhydria, and histological studies revealed abnormalities of the gastric epithelium, including moderate dilation of the gastric gland lumens and a reduction in the number of parietal cells. There was little evidence, however, that parietal cell viability was impaired. Ultrastructural analysis of AE2(-/-) gastric mucosa revealed abnormal parietal cell structure, with severely impaired development of secretory canaliculi and few tubulovesicles but normal apical microvilli. These results demonstrate that AE2 is essential for gastric acid secretion and for normal development of secretory canalicular and tubulovesicular membranes in mouse parietal cells. PMID- 15123621 TI - The aryl hydrocarbon receptor displaces p300 from E2F-dependent promoters and represses S phase-specific gene expression. AB - The environmental contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) causes a wide range of toxic, teratogenic, and carcinogenic effects. TCDD is a ligand for the aromatic hydrocarbon receptor (AHR), a ligand-activated transcription factor believed to be the primary mediator of these effects. Activation of the AHR by TCDD also elicits a variety of effects on cell cycle progression, ranging from proliferation to arrest. In this report, we have characterized further the role of the activated AHR in cell cycle regulation. In human mammary carcinoma MCF-7 and mouse hepatoma Hepa-1 cells, TCDD treatment decreased the number of cells in S phase and caused the accumulation of cells in G(1). In Hepa-1 cells, this effect correlated with the transcriptional repression of several E2F-regulated genes required for S phase progression. AHR-mediated gene repression was dependent on its interaction with retinoblastoma protein but was independent of its transactivation function because AHR mutants lacking DNA binding or transactivation domains repressed E2F-dependent expression as effectively as wild type AHR. Overexpression of p300 suppressed retinoblastoma protein-dependent gene repression, and this effect was reversed by TCDD. Chromatin immunoprecipitation assays showed that TCDD treatment caused the recruitment of AHR to E2F-dependent promoters and the concurrent displacement of p300. These results delineate a novel mechanism whereby the AHR, a known transcriptional activator, also mediates gene repression by pathways involving combinatorial interactions at E2F responsive promoters, leading to the repression of E2F-dependent, S phase specific genes. The AHR seems to act as an environmental checkpoint that senses exposure to environmental toxicants and responds by signaling cell cycle inhibition. PMID- 15123622 TI - Protein interactions involved in nuclear import of the Agrobacterium VirE2 protein in vivo and in vitro. AB - Agrobacterium, the only known organism capable of trans-kingdom DNA transfer, genetically transforms plants by transferring a segment of its DNA, T-DNA, into the nucleus of the host cell where it integrates into the plant genome. One of the central events in this genetic transformation process is nuclear import of the T-DNA molecule, which to a large degree is mediated by the bacterial virulence protein VirE2. VirE2 is distinguished by its nuclear targeting, which occurs only in plant but not in animal cells and is facilitated by the cellular VIP1 protein. The molecular mechanism of the VIP1 function is still unclear. Here, we used in vitro assays for nuclear import and quantification of protein protein interactions to directly demonstrate formation of ternary complexes between VirE2, VIP1, and a component of the cellular nuclear import machinery, karyopherin alpha. Our results indicate that VIP1 functions as a molecular bridge between VirE2 and karyopherin alpha, allowing VirE2 to utilize the host cell nuclear import machinery even without being directly recognized by its components. PMID- 15123623 TI - Neurokinin-B transcription in erythroid cells: direct activation by the hematopoietic transcription factor GATA-1. AB - The GATA family of transcription factors establishes genetic networks that control developmental processes including hematopoiesis, vasculogenesis, and cardiogenesis. We found that GATA-1 strongly activates transcription of the Tac-2 gene, which encodes proneurokinin-B, a precursor of neurokinin-B (NK-B). Neurokinins function through G protein-coupled transmembrane receptors to mediate diverse physiological responses including pain perception and the control of vascular tone. Whereas an elevated level of NK-B was implicated in pregnancy associated pre-eclampsia (Page, N. M., Woods, R. J., Gardiner, S. M., Lomthaisong, K., Gladwell, R. T., Butlin, D. J., Manyonda, I. T., and Lowry, P. J. (2000) Nature 405, 797-800), the regulation of NK-B synthesis and function are poorly understood. Tac-2 was expressed in normal murine erythroid cells and was induced upon ex vivo erythropoiesis. An estrogen receptor fusion to GATA-1 (ER GATA-1) and endogenous GATA-1 both occupied a region of Tac-2 intron-7, which contains two conserved GATA motifs. Genetic complementation analysis in GATA-1 null G1E cells revealed that endogenous GATA-2 occupied the same region of intron 7, and expression of ER-GATA-1 displaced GATA-2 and activated Tac-2 transcription. Erythroid cells did not express neurokinin receptors, whereas aortic and yolk sac endothelial cells differentially expressed neurokinin receptor subtypes. Since NK-B induced cAMP accumulation in yolk sac endothelial cells, these results suggest a new mode of vascular regulation in which GATA-1 controls NK-B synthesis in erythroid cells. PMID- 15123624 TI - Saccharomyces cerevisiae Hop1 zinc finger motif is the minimal region required for its function in vitro. AB - Saccharomyces cerevisiae meiosis-specific HOP1, which encodes a core component of synaptonemal complex, plays a key role in proper pairing of homologous chromosomes and processing of meiotic DNA double strand breaks. Isolation and analysis of hop1 mutants indicated that these functions require Cys(371) of Hop1 embedded in a region (residues 343-378) sharing homology to a zinc finger motif (ZnF). However, the precise biochemical function of Hop1, or its putative ZnF, in these processes is poorly understood. Our previous studies revealed that Hop1 is a DNA-binding protein, showed substantially higher binding affinity for G4 DNA, and enhances its formation. We report herein that ZnF appears to be sufficient for both zinc as well as DNA-binding activities. Molecular modeling studies suggested that Hop1 ZnF differs from the previously characterized natural ZnFs. The zinc-binding assay showed that the affinity for zinc is weaker for C371S ZnF mutant compared with the wild type (WT) ZnF. Analysis of CD spectra indicated that zinc and DNA induce substantial conformational changes in WT ZnF, but not in C371S ZnF mutant. The results from a number of different experimental approaches suggested that the DNA-binding properties of ZnF are similar to those of full length Hop1 and that interaction with DNA rich in G residues is particularly robust. Significantly, WT ZnF by itself, but not C371S mutant, was able to bind duplex DNA and promote interstitial pairing of DNA double helices via the formation of guanine quartets. Together, these results implicate a direct role for Hop1 in pairing of homologous chromosomes during meiosis. PMID- 15123625 TI - Transcriptional activity of peroxisome proliferator-activated receptor gamma is modulated by SUMO-1 modification. AB - Covalent modification of many transcription factors with SUMO-1 is emerging as a key role of trans-activational regulation. Here, we demonstrate that peroxisome proliferator-activated receptor (PPAR) gamma, which is a ligand-activated nuclear receptor, is modified by SUMO-1. Sumoylation of PPARgamma mainly occurs at a lysine residue within the activation function 1 domain. Furthermore, we show that the PIAS family proteins, PIAS1 and PIASxbeta, function as E3 ligases (ubiquitin protein isopeptide ligase) for PPARgamma. PPARgamma interacts directly with PIASxbeta in a ligand-independent manner. Analysis using a PPARgamma mutant with a disrupted sumoylation site shows that modification of PPARgamma by SUMO-1 represses its transcriptional activity. Interestingly, PIASxbeta and Ubc9 enhance the transcriptional activity of PPARgamma independent of PPARgamma sumoylation. Furthermore, PPARgamma ligand-induced apoptosis in a human hepatoblastoma cell line, HepG2, is significantly enhanced by ectopic production of the sumoylation mutant PPARgamma. These results suggest that the PPARgamma-dependent transactivation pathway seems to be modulated by SUMO-1 modification and may serve as a novel target for apoptosis-induction therapy in cancer cells. PMID- 15123626 TI - Cyclin-dependent kinase 5 associated with p39 promotes Munc18-1 phosphorylation and Ca(2+)-dependent exocytosis. AB - Cyclin-dependent kinase 5 (Cdk5) is a proline-directed serine/threonine protein kinase that requires association with a regulatory protein, p35 or p39, to form an active enzyme. Munc18-1 plays an essential role in membrane fusion, and its function is regulated by phosphorylation. We report here that both p35 and p39 were expressed in insulin-secreting beta-cells, where they exhibited individual subcellular distributions and associated with membranous organelles of different densities. Overexpression of Cdk5, p35, or p39 showed that Cdk5 and p39 augmented Ca(2+)-induced insulin exocytosis. Suppression of p39 and Cdk5, but not of p35, by antisense oligonucleotides selectively inhibited insulin exocytosis. Transient transfection of primary beta-cells with Munc18-1 templates mutated in potential Cdk5 or PKC phosphorylation sites, in combination with Cdk5 and the different Cdk5 activators, suggested that Cdk5/p39-promoted Ca(2+)-dependent insulin secretion from primary beta-cells by phosphorylating Munc18-1 at a biochemical step immediately prior to vesicle fusion. PMID- 15123627 TI - Regulation of CXC chemokine receptor 4-mediated migration by the Tec family tyrosine kinase ITK. AB - Chemokines are critical in controlling lymphocyte traffic and migration. The CXC chemokine CXCL12/SDF-1alpha interacts with its receptor CXCR4 to induce the migration of a number of different cell types. Although an understanding of the physiological functions of this chemokine is emerging, the mechanism by which it regulates T cell migration is still unclear. We show here that the Tec family kinase ITK is activated rapidly following CXCL12/SDF-1alpha stimulation, and this requires Src and phosphatidylinositol 3-kinase activities. ITK regulates the ability of CXCL12/SDF-1alpha to induce T cell migration as overexpression of wild type ITK-enhanced migration, and T cells lacking ITK exhibit reduced migration as well as adhesion in response to CXCL12/SDF-1alpha. Further analysis suggests that ITK may regulate CXCR4-mediated migration and adhesion by altering the actin cytoskeleton, as ITK null T cells were significantly defective in CXCL12/SDF-1a mediated actin polymerization. Our data suggest that ITK may regulate the ability of CXCR4 to induce T cell migration. PMID- 15123628 TI - High glucose down-regulates intercellular communication in retinal endothelial cells by enhancing degradation of connexin 43 by a proteasome-dependent mechanism. AB - Intercellular communication through gap junctions (GJIC) is most likely relevant to maintaining the integrity of the blood-retinal barrier. In this study, we investigated the mechanism whereby high glucose enhances degradation of connexin 43 (Cx43), thus contributing to a decrease in GJIC. The levels of Cx43 in bovine retinal endothelial cells exposed to high glucose (25 mm) decreased about 50% as compared with controls (5.5 mm glucose). Consistently, the half-life of the protein decreased from 2.3 to 1.9 h. The proteasome inhibitors MG132 and lactacystin prevented the loss of Cx43 induced by high glucose and extended Cx43 half-life. The amount of phosphorylated Cx43 increased in high glucose and after proteasome inhibition. Scrape-loading dye transfer experiments show that high glucose is associated to a decrease of 40% in GJIC. Significantly, this reduction can be reversed by proteasome inhibitors. The decrease in GJIC in cells exposed to high glucose is associated with a loss of Cx43 from the plasma membrane, as demonstrated by immunofluorescence and biotinylation of cell-surface proteins. Results indicate that increased phosphorylation of Cx43 under high glucose is the mechanism targeting Cx43 for degradation by a proteasome-dependent mechanism. Increased degradation of Cx43 and reduction of GJIC in high glucose may be of physiological importance by contributing to endothelial cell dysfunction associated with the breakdown of the blood-retinal barrier in diabetic retinopathy. PMID- 15123629 TI - Low levels of expression of leptin receptor at the cell surface result from constitutive endocytosis and intracellular retention in the biosynthetic pathway. AB - The leptin receptor is mainly localized in intracellular compartments in target tissues. To study the mechanisms leading to this intracellular localization, two main isoforms of leptin receptors, OB-Ra and OB-Rb, were expressed in HeLa cells. Both isoforms were localized at steady state in the trans-Golgi network, in endosomes, and to a lesser extent, at the cell surface. They turned over with a half-life of less than 2 h. Both isoforms of leptin receptors were constitutively endocytosed in a ligand-independent manner and degraded in lysosomes with no evidence of recycling to the cell surface or to the trans-Golgi network. The endocytosis was inhibited by the deletion of the cytoplasmic domain. Newly synthesized leptin receptors were partially retained in the Golgi complex or in a post-Golgi intracellular compartment. The transmembrane domain was found to be important for this intracellular retention in the biosynthetic pathway, whereas the cytoplasmic domain was not involved. The data suggest that the low levels of expression of leptin receptors at the cell surface results from partial retention in the biosynthetic pathway, coupled to constitutive removal from the plasma membrane via ligand-independent, constitutive endocytosis. PMID- 15123630 TI - Induction of heme oxygenase-1 inhibits NAD(P)H oxidase activity by down regulating cytochrome b558 expression via the reduction of heme availability. AB - Heme-oxygenase-1 (HO-1), the rate-limiting enzyme of heme degradation, has powerful anti-oxidant properties related to the production of the reactive oxygen species scavenger bilirubin. However, some data suggest that HO-1 could also inhibit the cellular production of reactive oxygen species. Therefore, we investigated whether the anti-oxidant properties of HO-1 could be mediated by modulation of the activity and/or expression of the heme-containing NAD(P)H oxidase, the main source of the superoxide anion (O(2)(-)) in phagocytic cells. Increasing HO-1 expression in RAW 264.7 macrophages effectively decreased NAD(P)H oxidase activity and expression of gp91(phox), its heme-containing catalytic component, because of deficient protein maturation and increased degradation. Loading cells with heme reversed the decrease in O(2)(-) production and gp91(phox) expression induced by HO-1 overexpression. Similar results were obtained in vivo in rat alveolar macrophages after pharmacological modulation of HO-1 expression or activity. These results show that a decrease in heme content due to HO-1 activation limits heme availability for maturation of the gp91(phox) subunit and assembly of the functional NAD(P)H oxidase. This study provides a new mechanism to explain HO-1 anti-oxidant properties. PMID- 15123631 TI - Positive regulation of phagocytosis by SIRPbeta and its signaling mechanism in macrophages. AB - SIRPbeta (signal-regulatory protein beta) is a transmembrane protein that is expressed in hematopoietic cells but whose functions are unknown. We have now cloned mouse SIRPbeta cDNA and have shown that the gene is expressed in various tissues in addition to cells of the macrophage lineage. Engagement of SIRPbeta by specific monoclonal antibodies promoted Fcgamma receptor-dependent or independent phagocytosis in mouse peritoneal macrophages. It also induced marked activation of MAPK and the upstream kinase MEK but weak activation of Akt. MEK inhibitors markedly blocked the promotion of phagocytosis by SIRPbeta, whereas an inhibitor of phosphoinositide 3-kinase partly blocked such response. In addition, inhibitors of myosin light chain kinase or of myosin ATPase blocked the promotion of phagocytosis by SIRPbeta. Furthermore, SIRPbeta induced the formation of filopodia and lamellipodia in macrophages as well as the translocation of activated MAPK to these structures. It also elicited tyrosine phosphorylation of DAP12, Syk, and SLP-76, and a Syk inhibitor blocked the promotion of phagocytosis and activation of MAPK by SIRPbeta. Our results suggest that engagement of SIRPbeta promotes phagocytosis in macrophages by inducing the tyrosine phosphorylation of DAP12, Syk, and SLP-76 and the subsequent activation of a MEK MAPK-myosin light chain kinase cascade. PMID- 15123632 TI - Phosphorylation of focal adhesion kinase at tyrosine 861 is crucial for Ras transformation of fibroblasts. AB - Although elevated expression and increased tyrosine phosphorylation of focal adhesion kinase (FAK) are crucial for tumor progression, the mechanism by which FAK promotes oncogenic transformation is unclear. We have therefore determined the role of FAK phosphorylation at tyrosine 861 in the oncogenic transformation of NIH3T3 fibroblasts. FAK phosphorylation at tyrosine 861 was increased in both constitutively H-Ras-transformed and H-Ras-inducible NIH3T3 cells, in parallel with cell transformation. However, H-Ras-inducible cells transfected with the nonphosphorylatable mutant FAK Y861F showed decreased migration/invasion, focus forming activity and anchorage-independent growth, compared with either wild-type or kinase-defective FAK. In contrast to unaltered FAK/Src activity, the association of FAK and p130(CAS) was decreased in FAK Y861F-transfected cells, and FAK phosphorylation at tyrosine 861 enhanced this association in vitro. Consistently, FAK Y861F-transfected cells were defective in activation of c-Jun NH(2)-terminal kinase and in expression of matrix metalloproteinase-9 during transformation. Taken together, these results strongly suggest that FAK phosphorylation at tyrosine 861 is crucial for H-Ras-induced transformation through regulation of the association of FAK with p130(CAS). PMID- 15123633 TI - 5'-3' RNA-RNA interaction facilitates cap- and poly(A) tail-independent translation of tomato bushy stunt virus mrna: a potential common mechanism for tombusviridae. AB - Tomato bushy stunt virus (TBSV) is the prototypical member of the genus Tombusvirus in the family Tombusviridae. The (+)-strand RNA genome of TBSV lacks both a 5' cap and a 3' poly(A) tail and instead contains a 3'-terminal RNA sequence that acts as a cap-independent translational enhancer (3' CITE). In this study, we have determined the RNA secondary structure of the translation-specific central segment of the 3' CITE, termed region 3.5 (R3.5). MFOLD structural modeling combined with solution structure mapping and comparative sequence analysis indicate that R3.5 adopts a branched structure that contains three major helices. Deletion and substitution studies revealed that two of these extended stem-loop (SL) structures are essential for 3' CITE activity in vivo. In particular, the terminal loop of one of these SLs, SL-B, was found to be critical for translation. Compensatory mutational analysis showed that SL-B functions by base pairing with another SL, SL3, in the 5' untranslated region of the TBSV genome. Thus, efficient translation of TBSV mRNA in vivo requires a 5'-3' RNA-RNA interaction that effectively circularizes the message. Similar types of interactions are also predicted to occur in TBSV subgenomic mRNAs between their 5' untranslated regions and the 3' CITE, and both genomic and subgenomic 5'-3' interactions are well conserved in all members of the genus Tombusvirus. In addition, a survey of other genera in Tombusviridae revealed the potential for similar 5'-3' RNA-RNA-based interactions in their viral mRNAs, suggesting that this mechanism extends throughout this large virus family. PMID- 15123634 TI - Requirement of histone deacetylase activity for signaling by STAT1. AB - STAT1 is a transcription factor that plays a crucial role in signaling by interferons (IFNs). In this study we demonstrated that inhibitors of histone deacetylase (HDAC) activity, butyrate, trichostatin A, and suberoylanilide hydroxamic acid, prevented IFNgamma-induced JAK1 activation, STAT1 phosphorylation, its nuclear translocation, and STAT1-dependent gene activation. Furthermore, we showed that silencing of HDAC1, HDAC2, and HDAC3 through RNA interference markedly decreased IFNgamma-driven gene activation and that overexpression of HDAC1, HDAC2, and HDAC3 enhanced STAT1-dependent transcriptional activity. Our data therefore established the essential role of deacetylase activity in STAT1 signaling. Induction of IRF-1 by IFNgamma requires functional STAT1 signaling and was abrogated by butyrate, trichostatin A, suberoylanilide hydroxamic acid, and STAT1 small interfering RNA. In contrast, silencing of STAT1 did not interfere with IFNgamma-induced expression of STAT2 and caspase-7, and HDAC inhibitors did not preclude IFNgamma-induced expression of STAT1, STAT2, and caspase-7, suggesting that HDAC inhibitors impede the expression of IFNgamma target genes whose expression depends on STAT1 but do not interfere with STAT1-independent signaling by IFNgamma. Finally, we showed that inhibitors of deacetylase activity sensitized colon cancer cells to IFNgamma induced apoptosis through cooperative negative regulation of Bcl-x expression, demonstrating that interruption of the balance between STAT1-dependent and STAT1 independent signaling significantly alters the biological activity of IFNgamma. PMID- 15123635 TI - Impact of replacement of D1 C-terminal alanine with glycine on structure and function of photosynthetic oxygen-evolving complex. AB - The C-terminal alanine 344 (Ala-344) in the D1 protein of photosystem II is conserved in all of the organisms performing oxygenic photosynthesis. A free alpha-COO(-) of Ala-344 has been proposed to be responsible for ligating the Mn cluster. Here, we constructed a mutant having D1 in which D1-Ala-344 was replaced with glycine (Gly) in cyanobacterium Synechocystis sp. PCC 6803. The effects of this minimal change in the side group from methyl to hydrogen on the properties of the oxygen-evolving complex were comprehensively investigated using purified core particles. The mutant grew photoautotrophically, and little change was observed in the protein composition of the oxygen-evolving core particles. The Gly-substituted oxygen-evolving complex showed small but normal S(2) multiline and enhanced g = 4.1 electron spin resonance signals and S(2)-state thermoluminescence bands with slightly elevated peak temperature. The Gly substitution resulted in distinct but relatively small changes in a few bands arising from the putative carboxylate ligand for the Mn cluster in the mid frequency (1800-1000 cm(-1)) S(2)/S(1) Fourier transform infrared difference spectrum. In contrast, the low frequency (670-350 cm(-1)) S(2)/S(1) Fourier transform infrared difference spectrum was markedly changed by the substitution. The results indicate that the internal structure of the Mn cluster and/or the interaction between the Mn cluster and its ligand are considerably altered by a simple change in the side group, from methyl to hydrogen, at the C-terminal of the D1 protein. PMID- 15123637 TI - The nucleoside derivative 5'-O-trityl-inosine (KIN59) suppresses thymidine phosphorylase-triggered angiogenesis via a noncompetitive mechanism of action. AB - Thymidine phosphorylase (TPase) catalyzes the reversible phosphorolysis of pyrimidine deoxynucleosides to 2-deoxy-d-ribose-1-phosphate and their respective pyrimidine bases. The enzymatic activity of TPase was found to be essential for its angiogenesis-stimulating properties. All of the previously described TPase inhibitors are either pyrimidine analogues that interact with the nucleoside binding site of the enzyme or modified purine derivatives that mimic the pyrimidine structure and either compete with thymidine or act as a multisubstrate (competitive) inhibitor. We now describe the inhibitory activity of the purine riboside derivative KIN59 (5'-O-tritylinosine) against human and bacterial recombinant TPase and TPase-induced angiogenesis. In contrast to previously described TPase inhibitors, KIN59 does not compete with the pyrimidine nucleoside or the phosphate-binding site of the enzyme but noncompetitively inhibits TPase when thymidine or phosphate is used as the variable substrate. In addition, KIN59 was far more active than other TPase inhibitors, previously tested by us, against TPase-induced angiogenesis in the chorioallantoic membrane assay. The observed anti-angiogenic effect of KIN59 was not accompanied by inflammation or any visible toxicity. Inosine did not inhibit the enzymatic or angiogenic activity of the enzyme, indicating that the 5'-O-trityl group in KIN59 is essential for the observed effects. In contrast with current concepts, our data indicate that the angiogenic activity of TPase is not solely directed through its functional nucleoside and phosphate-binding sites. Other regulatory (allosteric) site(s) in TPase may play an important role in the mechanism of TPase-triggered angiogenesis stimulation and apoptosis inhibition. Identification of these site(s) is important to obtain a better insight into the molecular role of TPase in the progression of cancer and angiogenic diseases. PMID- 15123636 TI - Specific role for p300/CREB-binding protein-associated factor activity in E2F1 stabilization in response to DNA damage. AB - E2F1, a member of the E2F family of transcription factors, plays a pivotal role in controlling both physiological cell-cycle progression and apoptotic cell death in response to DNA damage and oncogene activation. In response to genotoxic stresses, E2F1 is stabilized by signals that include ATM-dependent phosphorylation. We recently demonstrated that DNA damage induces also E2F1 acetylation, which is required for its recruitment onto apoptotic gene promoters. Here we show that E2F1 is stabilized in response to doxorubicin and cisplatin treatments even in the absence of either ATM-dependent phosphorylation or p53 and cAbl, two major transducers of DNA damage signaling. We found that acetylation of E2F1 is, instead, required to stabilize the protein in response to doxorubicin. Finally, we report that the formation of E2F1-p300/CREB-binding protein associated factor (P/CAF) complexes is preferentially induced in doxorubicin treated cells, and that P/CAF acetyltransferase (HAT), but not p300 HAT activity, is required for a significant E2F1 stabilization and accumulation. Our results unveil a differential role of P/CAF and p300 in acetylation-induced stabilization of E2F1, thus supporting a specific role for P/CAF HAT activity in E2F1-dependent apoptosis in response to DNA damage. PMID- 15123638 TI - BCL-2 translation is mediated via internal ribosome entry during cell stress. AB - The cellular response to stress involves a rapid inhibition of cap-dependent translation via multiple mechanisms, yet some translation persists. This residual translation may include proteins critical to the cellular stress response. BCL-2 is a key inhibitor of intrinsic apoptotic signaling. Its primary transcript contains a 1.45-kb 5'-untranslated region (UTR) including 10 upstream AUGs that may restrict translation initiation via cap-dependent ribosome scanning. Thus, we hypothesized that this 5'-UTR may contain an internal ribosome entry site (IRES) that facilitates BCL-2 translation, particularly during cell stress. Here we show that the BCL-2 5'-UTR demonstrated IRES activity both when translated in vitro and also when m(7)G-capped and polyadenylated mRNA was transiently transfected into 293T cells. The activity of this IRES in unstressed cells was approximately 6% the strength of the hepatitis C virus IRES but was induced 3-6-fold in a dose dependent manner following short term treatment with either etoposide or sodium arsenite. Thus, the IRES-mediated translation of BCL-2 may enable the cell to replenish levels of this critical protein during cell stress, when cap-dependent translation is repressed, thereby maintaining the balance between pro- and anti apoptotic BCL-2 family members in the cell and preventing unwarranted induction of apoptosis. PMID- 15123639 TI - A novel benzodiazepine increases the sensitivity of B cells to receptor stimulation with synergistic effects on calcium signaling and apoptosis. AB - Bz-423 is a 1,4-benzodiazepine with selective lymphotoxic properties and potent therapeutic activity against lupus-like disease in autoimmune mice. In NZB/W lupus-prone mice, Bz-423 specifically kills germinal center B cells, which are the cells that drive disease both in this model and in human systemic lupus erythematosus. In this report, the mechanistic basis for the selective action of Bz-423 is investigated. We show that Bz-423-induces superoxide as an immediate early response and that this reactive oxygen species is more effective as a second messenger death signal in B cells activated by B cell receptor stimulation compared with resting cells. As a result, low [Bz-423] that are not cytotoxic to non-stimulated cells kill stimulated cells in synergy with anti-immunoglobulin M antibodies. Subsequent experiments demonstrated that Bz-423 extends the rise in intracellular calcium that accompanies anti-immunoglobulin M stimulation, and this effect mediates the synergistic death response. Because B cell hyperactivation and altered calcium signaling is a distinguishing feature of autoreactive lymphocytes in lupus, the mechanism by which Bz-423 induces apoptosis preferentially targets disease-causing cells on the basis of their activation state. Thus, molecules like Bz-423 could form the basis for new and selective anti-lupus agents. PMID- 15123640 TI - Hyaluronan-CD44 interaction with Rac1-dependent protein kinase N-gamma promotes phospholipase Cgamma1 activation, Ca(2+) signaling, and cortactin-cytoskeleton function leading to keratinocyte adhesion and differentiation. AB - In this study we have investigated hyaluronan (HA)-CD44 interaction with protein kinase N-gamma (PKNgamma), a small GTPase (Rac1)-activated serine/threonine kinase in human keratinocytes. By using a variety of biochemical and molecular biological techniques, we have determined that CD44 and PKNgamma kinase (molecular mass approximately 120 kDa) are physically linked in vivo. The binding of HA to keratinocytes promotes PKNgamma kinase recruitment into a complex with CD44 and subsequently stimulates Rac1-mediated PKNgamma kinase activity. The Rac1 activated PKNgamma in turn increases threonine (but not serine) phosphorylation of phospholipase C (PLC) gamma1 and up-regulates PLCgamma1 activity leading to the onset of intracellular Ca(2+) mobilization. HA/CD44-activated Rac1-PKNgamma also phosphorylates the cytoskeletal protein, cortactin, at serine/threonine residues. The phosphorylation of cortactin by Rac1-PKNgamma attenuates its ability to cross-link filamentous actin in vitro. Further analyses indicate that the N-terminal antiparallel coiled-coil (ACC) domains of PKNgamma interact directly with Rac1 in a GTP-dependent manner. The binding of HA to CD44 induces PKNgamma association with endogenous Rac1 and its activity in keratinocytes. Transfection of keratinocytes with PKNgamma-ACCcDNA reduces HA-mediated recruitment of endogenous Rac1 to PKNgamma and blocks PKNgamma activity. These findings suggest that the PKNgamma-ACC fragment acts as a potent competitive inhibitor of endogenous Rac1 binding to PKNgamma in vivo. Most important, the PKNgamma-ACC fragment functions as a strong dominant-negative mutant that effectively inhibits HA/CD44-mediated PKNgamma phosphorylation of PLCgamma1 and cortactin as well as keratinocyte signaling (e.g. Ca(2+) mobilization and cortactin-actin binding) and cellular functioning (e.g. cell-cell adhesion and differentiation). Taken together, these findings strongly suggest that hyaluronan CD44 interaction with Rac1-PKNgamma plays a pivotal role in PLCgamma1-regulated Ca(2+) signaling and cortactin-cytoskeleton function required for keratinocyte cell-cell adhesion and differentiation. PMID- 15123641 TI - The structure of the Cys-rich terminal domain of Hydra minicollagen, which is involved in disulfide networks of the nematocyst wall. AB - The minicollagens found in the nematocysts of Hydra constitute a family of invertebrate collagens with unusual properties. They share a common modular architecture with a central collagen sequence ranging from 14 to 16 Gly-X-Y repeats flanked by polyproline/hydroxyproline stretches and short terminal domains that show a conserved cysteine pattern (CXXXCXXXCXXX-CXXXCC). The minicollagen cysteine-rich domains are believed to function in a switch of the disulfide connectivity from intra- to intermolecular bonds during maturation of the capsule wall. The solution structure of the C-terminal fragment including a minicollagen cysteine-rich domain of minicollagen-1 was determined in two independent groups by 1H NMR. The corresponding peptide comprising the last 24 residues of the molecule was produced synthetically and refolded by oxidation under low protein concentrations. Both presented structures are identical in their fold and disulfide connections (Cys2-Cys18, Cys6-Cys14, and Cys10-Cys19) revealing a robust structural motif that is supposed to serve as the polymerization module of the nematocyst capsule. PMID- 15123642 TI - Nitrate reductase activity is required for nitrate uptake into fungal but not plant cells. AB - The ability to transport net nitrate was conferred upon transformant cells of the non-nitrate-assimilating yeast Pichia pastoris after the introduction of two genes, one encoding nitrate reductase and the other nitrate transport. It was observed that cells of this lower eukaryote transformed with the nitrate transporter gene alone failed to display net nitrate transport despite having the ability to produce the protein. In addition, loss-of-function nitrate reductase mutants isolated from several nitrate-assimilating fungi appeared to be unable to accumulate nitrate. Uptake assays using the tracer (13)NO(3)(-) showed that nitrate influx is negligible in cells of a nitrate reductase null mutant. In parallel studies using a higher eukaryotic plant, Arabidopsis thaliana, loss-of function nitrate reductase strains homozygous for both NIA1 insertion and NIA2 deletion were found to have no detectable nitrate reductase mRNA or nitrate reductase activity but retained the ability to transport nitrate. The reasons for these fundamental differences in nitrate transport into the cells of representative members of these two eukaryotic kingdoms are discussed. PMID- 15123643 TI - Crystal structure of PapA5, a phthiocerol dimycocerosyl transferase from Mycobacterium tuberculosis. AB - Polyketide-associated protein A5 (PapA5) is an acyltransferase that is involved in production of phthiocerol and phthiodiolone dimycocerosate esters, a class of virulence-enhancing lipids produced by Mycobacterium tuberculosis. Structural analysis of PapA5 at 2.75-A resolution reveals a two-domain structure that shares unexpected similarity to structures of chloramphenicol acetyltransferase, dihydrolipoyl transacetylase, carnitine acetyltransferase, and VibH, a non ribosomal peptide synthesis condensation enzyme. The PapA5 active site includes conserved histidine and aspartic acid residues that are critical to PapA5 acyltransferase activity. PapA5 catalyzes acyl transfer reactions on model substrates that contain long aliphatic carbon chains, and two hydrophobic channels were observed linking the PapA5 surface to the active site with properties consistent with these biochemical activities and substrate preferences. An additional alpha helix not observed in other acyltransferase structures blocks the putative entrance into the PapA5 active site, indicating that conformational changes may be associated with PapA5 activity. PapA5 represents the first structure solved for a protein involved in polyketide synthesis in Mycobacteria. PMID- 15123644 TI - Conserved amino acid residues in the COOH-terminal tail are indispensable for the correct folding and localization and enzyme activity of neutral ceramidase. AB - Several lines of evidence suggest that neutral ceramidase is involved in the regulation of ceramide-mediated signaling. Recently, the enzymes from mouse and rat were found to be localized at plasma membranes as a type II integral membrane protein, occasionally being detached from the cells after proteolytic processing of the NH(2)-terminal anchoring region (Tani, M., Iida, H., and Ito, M. (2003) J. Biol. Chem. 278, 10523-10530). We report here that conserved hydrophobic amino acid residues in the COOH-terminal tail are indispensable for the correct folding and localization, and enzyme activity of neutral ceramidase. Truncation of four, but not three, amino acid residues from the COOH terminus of rat neutral ceramidase resulted in a complete loss of enzyme activity as well as cell surface expression in HEK293 cells. Point mutation analysis revealed that Ile(758), the 4(th) amino acid residue from the COOH terminus, and Phe(756) are essential for the enzyme to function. The truncated and mutated enzymes were found to be retained in the endoplasmic reticulum (ER) and rapidly degraded without transportation to the Golgi apparatus. Treatment of the cells expressing the aberrant COOH-terminal enzyme with MG-132, a specific inhibitor for the proteasome, increased the accumulation of the enzyme in the ER, indicating that the misfolded enzyme was degraded by the proteasome. It was also found that the COOH-terminal tail was indispensable for the enzyme activity and correct folding of the prokaryote ceramidase from Pseudomonas aeruginosa, indicating that the importance of the COOH-terminal tail of the enzyme has been preserved through evolution. PMID- 15123645 TI - Early painful diabetic neuropathy is associated with differential changes in tetrodotoxin-sensitive and -resistant sodium channels in dorsal root ganglion neurons in the rat. AB - Diabetic neuropathy is a common form of peripheral neuropathy, yet the mechanisms responsible for pain in this disease are poorly understood. Alterations in the expression and function of voltage-gated tetrodotoxin-resistant (TTX-R) sodium channels have been implicated in animal models of neuropathic pain, including models of diabetic neuropathy. We investigated the expression and function of TTX sensitive (TTX-S) and TTX-R sodium channels in dorsal root ganglion (DRG) neurons and the responses to thermal hyperalgesia and mechanical allodynia in streptozotocin-treated rats between 4-8 weeks after onset of diabetes. Diabetic rats demonstrated a significant reduction in the threshold for escape from innocuous mechanical pressure (allodynia) and a reduction in the latency to withdrawal from a noxious thermal stimulus (hyperalgesia). Both TTX-S and TTX-R sodium currents increased significantly in small DRG neurons isolated from diabetic rats. The voltage-dependent activation and steady-state inactivation curves for these currents were shifted negatively. TTX-S currents induced by fast or slow voltage ramps increased markedly in neurons from diabetic rats. Immunoblots and immunofluorescence staining demonstrated significant increases in the expression of Na(v)1.3 (TTX-S) and Na(v) 1.7 (TTX-S) and decreases in the expression of Na(v) 1.6 (TTX-S) and Na(v)1.8 (TTX-R) in diabetic rats. The level of serine/threonine phosphorylation of Na(v) 1.6 and In Na(v)1.8 increased in response to diabetes. addition, increased tyrosine phosphorylation of Na(v)1.6 and Na(v)1.7 was observed in DRGs from diabetic rats. These results suggest that both TTX-S and TTX-R sodium channels play important roles and that differential phosphorylation of sodium channels involving both serine/threonine and tyrosine sites contributes to painful diabetic neuropathy. PMID- 15123646 TI - Janus kinase (Jak) subcellular localization revisited: the exclusive membrane localization of endogenous Janus kinase 1 by cytokine receptor interaction uncovers the Jak.receptor complex to be equivalent to a receptor tyrosine kinase. AB - The Janus kinases are considered to be cytoplasmic kinases that constitutively associate with the cytoplasmic region of cytokine receptors, and the Janus kinases (Jaks) are crucial for cytokine signal transduction. We investigated Jak1 localization using subcellular fractionation techniques and fluorescence microscopy (immunofluorescence and yellow fluorescent protein-tagged Jaks). In the different experimental approaches we found Jak1 (as well as Jak2 and Tyk2) predominantly located at membranes. In contrast to previous reports we did not observe Jak proteins in significant amounts within the nucleus or in the cytoplasm. The cytoplasmic localization observed for the Jak1 mutant L80A/Y81A, which is unable to associate with cytokine receptors, indicates that Jak1 does not have a strong intrinsic membrane binding potential and that only receptor binding is crucial for the membrane recruitment. Finally we show that Jak1 remains a membrane-localized protein after cytokine stimulation. These data strongly support the hypothesis that cytokine receptor.Janus kinase complexes can be regarded as receptor tyrosine kinases. PMID- 15123647 TI - Characterization of structural determinants of granzyme B reveals potent mediators of extended substrate specificity. AB - Granzymes are trypsin-like serine proteases mediating apoptotic cell death that are composed of two genetically distinct subfamilies: granzyme A-like proteases resemble trypsin in their active site architecture, while granzyme B-like proteases are quite distinct. Granzyme B prefers substrates containing P4 to P1 amino acids Ile/Val, Glu/Met/Gln, Pro/Xaa, and aspartic acid N-terminal to the proteolytic cleavage. By investigating the narrow extended specificity of the granzyme B-like proteases the mediators of their unique specificity are being defined. The foci of this study were the structural determinants Ile99, Tyr174, Arg192, and Asn218. Even modest mutations of these residues resulted in unique extended specificity profiles as determined using combinatorial substrate libraries and individual fluorogenic substrates. As with other serine proteases, Ile99 completely defines and predicts P2 specificity, primarily through the binding constant Km. Asn218 variants have minor effects alone but in combination with mutations at Arg192 and Ile99 alter P2 through P4 extended specificity. For each variant, the activity on its cognate substrate was equal to that of granzyme B for the same substrate. Thus, mutations at these determinants change extended selectivity preferentially over catalytic power. Additionally Asn218 variants result in increased activity on the wild type substrate, while the N218A/I99A variant disrupts the additivity between P2 and P4 specificity. This defines Asn218 not only as a determinant of specificity but also as a structural component required for P2 and P4 independence. This study confirms four determinants of granzyme B extended substrate specificity that constitute a canon applicable to the study of the remaining family members. PMID- 15123648 TI - Functional characterization of a trafficking-defective HCN4 mutation, D553N, associated with cardiac arrhythmia. AB - Hyperpolarization-activated cyclic nucleotide-gated channel 4 gene HCN4 is a pacemaker channel that plays a key role in automaticity of sinus node in the heart, and an HCN4 mutation was reported in a patient with sinus node dysfunction. Expression of HCN4 in the heart is, however, not confined to the sinus node cells but is found in other tissues, including cells of the conduction system. On the other hand, mutations in another cardiac ion channel gene, SCN5A, also cause sinus node dysfunction as well as other cardiac arrhythmias, including long QT syndrome, Brugada syndrome, idiopathic ventricular fibrillation, and progressive cardiac conduction disturbance. These observations imply that HCN4 abnormalities may be involved in the pathogenesis of various arrhythmias, similar to the SCN5A mutations. In this study, we analyzed patients suffering from sinus node dysfunction, progressive cardiac conduction disease, and idiopathic ventricular fibrillation for mutations in HCN4. A missense mutation, D553N, was found in a patient with sinus node dysfunction who showed recurrent syncope, QT prolongation in electrocardiogram, and polymorphic ventricular tachycardia, torsade de pointes. In vitro functional study of the D553N mutation showed a reduced membranous expression associated with decreased If currents because of a trafficking defect of the HCN4 channel in a dominant-negative manner. These data suggest that the loss of function of HCN4 is associated with sinus nodal dysfunction and that a consequence of pacemaker channel abnormality might underlie clinical features of QT prolongation and polymorphic ventricular tachycardia developed under certain conditions. PMID- 15123649 TI - SREBP-1c mediates the insulin-dependent hepatic glucokinase expression. AB - The regulation of hepatic glucose metabolism is important in glucose homeostasis, and liver glucokinase (LGK) plays a central role in this process. Hepatic glucokinase expression is known to be regulated by insulin. Recently it has been suggested that sterol regulatory element binding protein-1c (SREBP-1c) mediates the action of insulin on LGK transcription; however, the precise mechanism is not, to date, well known. In the present study, we identified two functional SREBP-1c response elements, SREa and SREb, in the rat LGK promoter. SREBP-1c could bind to these SREs and activate the LGK promoter, and insulin activated the LGK promoter in Alexander cells. The physical interaction between the protein and SREs of the LGK promoter in vivo was also confirmed. Insulin selectively increased SREBP-1c and LGK expression in primary hepatocytes. Adenoviral expression of SREBP-1c stimulated LGK expression, and the dominant negative mutant of SREBP-1c blocked the increased gene expression of LGK by insulin and SREBP-1c. A chromatin immunoprecipitation assay using primary hepatocytes showed increased binding of SREBP-1 to SREs of the LGK promoter by insulin. PMID- 15123650 TI - Differential regulation of human and mouse orphan nuclear receptor small heterodimer partner promoter by sterol regulatory element binding protein-1. AB - Small heterodimer partner (SHP; NR0B2) is an unusual orphan nuclear receptor that lacks a conventional DNA-binding domain and acts as a modulator of transcriptional activities of a number of nuclear receptors. Herein, we report that the human SHP promoter (hSHP) is activated by sterol regulatory element binding protein-1 (SREBP-1), which regulates the expression of various genes involved in cholesterol and fatty acid synthesis. Overexpression of SREBP-1 activated the human but not mouse SHP promoter, although SREBP-2 had little effect on the SHP promoter in CV-1 cells. Serial deletion reporter assays revealed that SREBP-1-responsive region is located within the sequences from -243 to -120 bp in the hSHP promoter. DNase I footprinting, gel shift assays, and chromatin immunoprecipitation assays demonstrated that SREBP-1 binds directly to the hSHP promoter. Site-directed mutagenesis made it clear that the hSHP promoter activation by SREBP-1 is mostly mediated by the SRE1 (-186 to -195 bp) in the hSHP promoter, which is not conserved in the mouse SHP promoter. Moreover, adenovirus-mediated overexpression of SREBP-1c/ADD-1 induced SHP mRNA expression and repressed CYP7A1 expression in HepG2 cells. Finally, we found that a four nucleotide deletion (-195CT-GAdel) in the hSHP promoter, which is reported to be associated with altered body weight and insulin secretion in human, coincides with the SRE1. This mutation strongly decreased both basal and SREBP-1 dependent activities of the hSHP promoter, because of the reduced binding of SREBP-1 to the mutated SRE1. Overall, our results demonstrate a differential regulation of human and mouse SHP promoters by SREBP-1. We propose a possible role of SREBP-1 in the species differential regulation of cholesterol and bile acid homeostasis via a novel mechanism of up-regulation of the hSHP gene expression. PMID- 15123651 TI - hXRCC2 enhances ADP/ATP processing and strand exchange by hRAD51. AB - The assembly of bacterial RecA, and its human homolog hRAD51, into an operational ADP/ATP-regulated DNA-protein (nucleoprotein) filament is essential for homologous recombination repair (HRR). Yet hRAD51 lacks the coordinated ADP/ATP processing exhibited by RecA and is less efficient in HRR reactions in vitro. In this study, we demonstrate that hXRCC2, one of five other poorly understood non redundant human mitotic RecA homologs (hRAD51B, hRAD51C, hRAD51D, hXRCC2, and hXRCC3), stimulates hRAD51 ATP processing. hXRCC2 also increases hRAD51-mediated DNA unwinding and strand exchange activities that are integral for HRR. Although there does not seem to be a long-lived interaction between hXRCC2 and hRAD51, we detail a strong adenosine nucleotide-regulated interaction between the hXRCC2 hRAD51D heterodimer and hRAD51. These observations begin to elucidate the separate and specialized functions of the human mitotic RecA homologs that enable an efficient nucleoprotein filament required for HRR. PMID- 15123652 TI - The rat leukocyte-type 12-lipoxygenase exhibits an intrinsic hepoxilin A3 synthase activity. AB - Hepoxilins are biologically relevant eicosanoids formed via the 12-lipoxygenase pathway of the arachidonic acid cascade. Although these eicosanoids exhibit a myriad of biological activities, their biosynthetic mechanism has not been investigated in detail. We examined the arachidonic acid metabolism of RINm5F rat insulinoma cells and found that they constitutively express a leukocyte-type 12S lipoxygenase. Moreover, we observed that RINm5F cells exhibit an active hepoxilin A(3) synthase that converts exogenous 12S-HpETE (12S-5Z,8-Z,10E,14Z-12 hydro(pero)xy-eicosa-5,8,10,14-tetraenoic acid) or arachidonic acid predominantly to hepoxilin A(3). 12S-lipoxygenase and hepoxilin A(3) synthase activities were co-localized in the cytosol; immunoprecipitation with an anti-12S-lipoxygenase antibody co-precipitated the two catalytic activities. These data suggested that hepoxilin A(3) synthase activity may be considered an intrinsic catalytic property of the leukocyte-type 12S-lipoxygenase. To test this hypothesis we cloned the leukocyte-type 12S-LOX from RINm5F cells, expressed it in Pichia pastoris, and found that the recombinant enzyme exhibited both 12S-lipoxygenase and hepoxilin A(3) synthase activities. The recombinant human platelet-type 12S lipoxygenase and the porcine leukocyte-type 12S-lipoxygenase also exhibited hepoxilin A(3) synthase activity. In contrast, the native rabbit reticulocyte type 15S-lipoxygenase did not convert 12S-HpETE to hepoxilin isomers. These data suggest that the positional specificity of lipoxygenases may be crucial for this catalytic function. This hypothesis was confirmed by site-directed mutagenesis studies that altered the positional specificity of the rat leukocyte-type 12S- and the rabbit reticulocyte-type 15-lipoxygenase. In summary, it may be concluded that naturally occurring 12S-lipoxygenases exhibit an intrinsic hepoxilin A(3) synthase activity that is minimal in lipoxygenase isoforms with different positional specificity. PMID- 15123653 TI - Notch oncoproteins depend on gamma-secretase/presenilin activity for processing and function. AB - During normal development Notch receptor signaling is important in regulating numerous cell fate decisions. Mutations that truncate the extracellular domain of Notch receptors can cause aberrant signaling and promote unregulated cell growth. We have examined two types of truncated Notch oncoproteins that arise from proviral insertion into the Notch4 gene (Notch4/int-3) or a chromosomal translocation involving the Notch1 gene (TAN-1). Both Notch4/int-3 and TAN-1 oncoproteins lack most or all of their ectodomain. Normal Notch signaling requires gamma-secretase/presenilin-mediated proteolytic processing, but whether Notch oncoproteins are also dependent on gamma-secretase/presenilin activity is not known. We demonstrate that Notch4/int-3-induced activation of the downstream transcription factor, CSL, is abrogated in cells deficient in presenilins or treated with a pharmacological inhibitor of gamma-secretase/presenilins. Furthermore, we find that both Notch4/int-3 and TAN-1 accumulate at the cell surface, where presenilin-dependent cleavage occurs, when gamma secretase/presenilin activity is inhibited. gamma-Secretase/presenilin inhibition effectively blocks cellular responses to Notch4/int-3, but not TAN-1, apparently because some TAN-1 polypeptides lack transmembrane domains and do not require gamma-secretase/presenilin activity for nuclear access. These studies highlight potential uses and limitations of gamma-secretase/presenilin inhibitors in targeted therapy of Notch-related neoplasms. PMID- 15123654 TI - Single nucleotide polymorphism (-468 Gly to A) at the promoter region of SREBP-1c associates with genetic defect of fructose-induced hepatic lipogenesis [corrected]. AB - To evaluate the genetic susceptibility to metabolic disorders induced by high fructose diet, we investigated the metabolic characteristics in 10 strains of inbred mice and found that they were separated into CBA and DBA groups according to the response to high fructose diet. The hepatic mRNA expression of the sterol regulatory element-binding protein-1 (SREBP-1) in CBA/JN was remarkably enhanced by high fructose diet but not in DBA/2N. Similar results were observed in primary hepatocytes after exposure to fructose. The nucleotide sequence at -468 bp from the putative starting point of the SREBP-1c gene was adenine in the DBA group while it was guanine in the CBA group. In hepatocytes from CBA/JN, the activity of CBA-SREBP-1c promoter was significantly increased by 2.4- and 2.2-fold, in response to 30 mm fructose or 10 nm insulin, respectively, whereas the activity of DBA-SREBP-1c promoter responded to insulin but not to fructose. In hepatocytes from DBA/2N, both types of SREBP-1c promoter activities in response to insulin were attenuated. Furthermore, electrophoretic mobility shift assay revealed an unidentified nuclear protein bound to the oligonucleotides made from the region between -453 to -480 bp of the SREBP-1c promoter of CBA/JN but not to the probe from DBA/2N. Thus, in DBA/2N, the reduced mRNA expression of SREBP-1 after fructose refeeding appeared to associate with two independent mechanisms, 1). loss of binding of unidentified proteins to the region between -453 to -480 bp of the SREBP-1c promoter and 2). impaired insulin stimulation of SREBP-1c promoter activity. PMID- 15123655 TI - Genetic interactions between Brca1 and Gadd45a in centrosome duplication, genetic stability, and neural tube closure. AB - GADD45a is a transcription target of the breast tumor suppressor gene BRCA. It was recently shown that mouse embryonic fibroblast cells carrying a targeted deletion of exon 11 of Brca1 (Brca1(Delta11/Delta11)) or a Gadd45A-null mutation (Gadd45a(-/-)) suffer centrosome amplification. To study genetic interactions between these genes during centrosome duplication, we generated Brca1(Delta11/Delta)(11)Gadd45a(-/-) mice by crossing each mutant. We found that all Brca1(Delta11/Delta11)Gadd45a(-/-) embryos at embryonic days 9.5-10.5 were exencephalic and exhibited a high incidence of apoptosis accompanied by altered levels of BAX, BCL-2, and p53. The trigger for these events is likely the genetic instability arising from centrosome amplification that is associated, at least in part, with decreased expression of the NIMA-related kinase NEK2. We demonstrate that small interfering RNA-mediated suppression of Brca1 decreased Nek2 more dramatically in Gadd45a(-/-) cells than in wild-type cells and, conversely, that overexpression of Brca1 and/or Gadd45a up-regulated transcription of Nek2. Furthermore, we show that overexpression of Nek2 in Brca1-specific small interfering RNA-treated wild-type and Gadd45a(-/-) cells repressed abnormal centrosome amplification. These observations suggest that NEK2 plays a role in mediating the actions of BRCA1 and GADD45a in regulating centrosome duplication and in maintaining genetic stability. PMID- 15123656 TI - Characterization of sugar binding by osteoclast inhibitory lectin. AB - Osteoclast inhibitory lectin (OCIL) is a membrane-bound C-type lectin that blocks osteoclast differentiation and, via binding to its cognate receptor NKRP1D, inhibits natural killer cell-mediated cytotoxicity. OCIL is a member of the natural killer cell receptor C-type lectin group that includes CD69 and NKRP1D. We investigated carbohydrate binding of soluble recombinant human and mouse OCIL in enzyme-linked immunosorbent assay-based assays. OCIL bound immobilized high molecular weight sulfated glycosaminoglycans, including fucoidan, lambda carrageenan, and dextran sulfate, but not unsulfated dextran or sialated hyaluronic acid. Carbohydrate binding was Ca(2+)-independent. Binding of immobilized low molecular weight glycosaminoglycans, including chondroitin sulfate (A, B, and C forms) and heparin, was not observed. However, the soluble forms of these low molecular weight glycosaminoglycans competed for OCIL binding of immobilized fucoidan (as did soluble fucoidan, dextran sulfate, and lambda carrageenan), indicating that OCIL does recognize these carbohydrates. Inhibition constants for chondroitin sulfate A and heparin binding were 380 and 5 nm, respectively. Immobilized and soluble monosaccharides did not bind OCIL. The presence of saturating levels of fucoidan, dextran sulfate, and lambda carrageenan did not affect OCIL inhibition of osteoclast formation. The fucoidan binding lectins Ulex europaeus agglutinin I and Anguilla anguilla agglutinin did not block osteoclast formation or affect the inhibitory action of OCIL. Although the osteoclast inhibitory action of OCIL is independent of sugar recognition, we have found that OCIL, a lectin widely distributed, but notably localized in bone, skin, and other connective tissues, binds a range of physiologically important glycosaminoglycans, and this property may modulate OCIL actions upon other cells. PMID- 15123657 TI - Osteopenia in Siah1a mutant mice. AB - Siah1a has been implicated in numerous signaling pathways because of its ability to induce ubiquitin-mediated degradation of many protein substrates. Siah1a knockout mice are growth-retarded, exhibit early lethality, and display spermatogenic defects. In this study we identified a striking low bone volume phenotype in these mice (trabecular bone volume was halved compared with wild type mice), linking Siah1a to bone metabolism for the first time. Markers of bone formation, including osteoblast numbers and osteoid volume, were decreased by up to 40%, whereas the number of osteoclasts was more than doubled in Siah1a mutant mice. However, ex vivo osteoclast formation occurs normally and hematopoietic osteoclast progenitor cell types were present in normal numbers in Siah1a mutant mice. Moreover, adoptive transfer of Siah1a mutant bone marrow into wild type mice failed to reproduce the osteopenia or increased osteoclast numbers observed in mutant mice. Although ex vivo osteoblast colony formation was normal in Siah1a mutant mice, mineralization from these cells was elevated in cultures from Siah1a mutant mice, which may explain the reduction in osteoid volume seen in vivo. These findings suggest that although Siah1a is clearly essential for normal bone metabolism, the bone defect in Siah1a mutant mice is not due to cell-autonomous requirements for Siah1a in osteoblast or osteoclast formation. We propose that bone metabolism defects in Siah1a mutant mice are secondary to an alteration in an unidentified systemic, paracrine, or metabolic factor in these mice. PMID- 15123658 TI - Localization of a cryptic binding site for tenascin on fibronectin. AB - Fibronectin and tenascin are large extracellular matrix proteins that interact with each other and with integrin receptors to regulate cell growth and movement. They are both modular proteins composed of independently folded domains (modules) that are arranged in linear fashion. Fibronectin is a covalent dimer and tenascin is a hexamer. The site on tenascin to which fibronectin binds has been localized to type III modules 3-5. In this study we use surface plasmon resonance to examine the interaction between various fragments of fibronectin and tenascin to further characterize and localize the binding sites. We found that tenascin fragments that contain type III modules 3-5 bind primarily to the N-terminal 29 kDa hep-1/fib-1 domain, which contains the first five type I modules of fibronectin. The dissociation constant, K(d), is approximately 1 microm. The binding site on fibronectin appears to be cryptic in the whole molecule in solution but is exposed on the proteolytic fragments and probably when fibronectin is in the extended conformation. PMID- 15123659 TI - Identification of the region in Cdc42 that confers the binding specificity to activated Cdc42-associated kinase. AB - The Rho family small G-protein Cdc42 has been implicated in a diversity of biological functions. Multiple downstream effectors have been identified. How Cdc42 discriminates the interaction with its multiple downstream effectors is not known. Activated Cdc42-associated tyrosine kinase (ACK) is a very specific effector of Cdc42. To delineate the Cdc42 signaling pathway mediated by ACK, we set about to identify the specific ACK-binding region in Cdc42. We utilized TC10, another member of the Rho family of G-proteins that is 66.7% identical to Cdc42, to construct TC10/Cdc42 chimeras for screening the specific ACK-binding region in Cdc42. A region between switch I and switch II has been identified as the specific ACK-binding (AB) region. The replacement of the AB region with the corresponding region in TC10 resulted in the complete loss of ACK-binding ability but did not affect the binding to WASP, suggesting that the AB region confers the binding specificity to ACK. On the other hand, replacement of the corresponding region of TC10 with the AB region enabled TC10 to acquire ACK-binding ability. Eight residues are different between the AB region and the corresponding region of TC10. The mutational analysis indicated that all eight residues contribute to the binding to ACK2. The assays for the Cdc42-mediated activation of ACK2 indicated that the AB region is essential for Cdc42 to activate ACK2 in cells. Thus, our studies have defined a specific ACK-binding region in Cdc42 and have provided a molecular basis for generating ACK binding-defective mutants of Cdc42 to delineate ACK-mediated signaling pathway. PMID- 15123660 TI - Specificity of a soluble UDP-galactose: fucoside alpha1,3-galactosyltransferase that modifies the cytoplasmic glycoprotein Skp1 in Dictyostelium. AB - Skp1 is an adaptor-like protein in E3(SCF)-ubiquitin ligases and other multiprotein complexes of the cytoplasm and nucleus. In Dictyostelium, Skp1 is modified by an unusual pentasaccharide containing a Galalpha1-Fuc linkage, whose formation is examined here. A cytosolic extract from Dictyostelium was found to yield, after 2400-fold purification, an activity that could transfer Gal from UDP Gal to both a Fuc-terminated glycoform of Skp1 and synthetic Fuc conjugates in the presence of Mn(2+) and dithiothreitol. The microsomal fraction was devoid of activity. The linkage formed was Galalpha1,3Fuc based on co-chromatography with only this synthetic isomer conjugate, and sensitivity to alpha1,3/6 galactosidase. Skp1 exhibited an almost 1000-fold lower K(m) and 35-fold higher V(max) compared with a simple alpha-fucoside, but this advantage was abolished by denaturation or alkylation of Cys residues. A comparison of a complete series of synthetic glycosides representing the non-reducing terminal mono-, di-, and trisaccharides of Skp1 revealed, surprisingly, that the disaccharide is most active owing primarily to a V(max) advantage, but still much less active than Skp1 itself because of a K(m) difference. These findings indicate that alpha GalT1 is a cytoplasmic enzyme whose modification of Skp1 requires proper presentation of the terminal acceptor disaccharide by a folded Skp1 polypeptide, which correlates with previous evidence that the Galalpha1,3Fuc linkage is deficient in expressed mutant Skp1 proteins. PMID- 15123661 TI - A conserved motif for the transport of G protein-coupled receptors from the endoplasmic reticulum to the cell surface. AB - The structural determinants for the export trafficking of G protein-coupled receptors are poorly defined. In this report, we determined the role of carboxyl termini (CTs) of alpha2B-adrenergic receptor (AR) and angiotensin II type 1A receptor (AT1R) in their transport from the endoplasmic reticulum (ER) to the cell surface. The alpha2B-AR and AT1R mutants lacking the CTs were completely unable to transport to the cell surface and were trapped in the ER. Alanine scanning mutagenesis revealed that residues Phe436 and Ile433-Leu444 in the CT were required for alpha2B-AR export. Insertion or deletion between Phe436 and Ile443-Leu444 as well as Ile443-Leu444 mutation to FF severely disrupted alpha2B AR transport, indicating there is a defined spatial requirement, which is essential for their function as a single motif regulating receptor transport from the ER. Furthermore, the carboxyl-terminally truncated as well as Phe436 and Ile443-Leu444 mutants were unable to bind ligand and the alpha2B-AR CT conferred its transport properties to the AT1R mutant without the CT in a Phe436-Ile443 Leu444-dependent manner. These data suggest that the Phe436 and Ile443-Leu444 may be involved in both proper folding and export from the ER of the receptor. Similarly, residues Phe309 and Leu316-Leu317 in the CT were identified as essential for AT1R export. The sequence F(X)6LL (where X can be any residue, and L is leucine or isoleucine) is highly conserved in the membrane-proximal CTs of many G protein-coupled receptors and may function as a common motif mediating receptor transport from the ER to the cell surface. PMID- 15123662 TI - Antagonistic cross-talk between Rac and Cdc42 GTPases regulates generation of reactive oxygen species. AB - Cross-talk between Rho GTPase family members (Rho, Rac, and Cdc42) plays important roles in modulating and coordinating downstream cellular responses resulting from Rho GTPase signaling. The NADPH oxidase of phagocytes and nonphagocytic cells is a Rac GTPase-regulated system that generates reactive oxygen species (ROS) for the purposes of innate immunity and intracellular signaling. We recently demonstrated that NADPH oxidase activation involves sequential interactions between Rac and the flavocytochrome b(558) and p67(phox) oxidase components to regulate electron transfer from NADPH to molecular oxygen. Here we identify an antagonistic interaction between Rac and the closely related GTPase Cdc42 at the level of flavocytochrome b(558) that regulates the formation of ROS. Cdc42 is unable to stimulate ROS formation by NADPH oxidase, but Cdc42, like Rac1 and Rac2, was able to specifically bind to flavocytochrome b(558) in vitro. Cdc42 acted as a competitive inhibitor of Rac1- and Rac2-mediated ROS formation in a recombinant cell-free oxidase system. Inhibition was dependent on the Cdc42 insert domain but not the Switch I region. Transient expression of Cdc42Q61L inhibited ROS formation induced by constitutively active Rac1 in an NADPH oxidase-expressing Cos7 cell line. Inhibition of Cdc42 activity by transduction of the Cdc42-binding domain of Wiscott-Aldrich syndrome protein into human neutrophils resulted in an enhanced fMetLeuPhe-induced oxidative response, consistent with inhibitory cross-talk between Rac and Cdc42 in activated neutrophils. We propose here a novel antagonism between Rac and Cdc42 GTPases at the level of the Nox proteins that modulates the generation of ROS used for host defense, cell signaling, and transformation. PMID- 15123663 TI - Colon carcinoma cell growth is associated with prostaglandin E2/EP4 receptor evoked ERK activation. AB - Cyclooxygenase (COX) and its prostanoid metabolites have been implicated in the control of cell survival; however, their role as mitogens remains undefined. To better understand the role of prostanoids on cell growth, we used mouse colon adenocarcinoma (CT26) cells to investigate the role of prostaglandin E(2) (PGE(2)) in cell proliferation. CT26 cells express both COX1 and COX2 and metabolize arachidonic acid to PGE(2.) Treatment with indomethacin, or COX selective inhibitors, prevents PGE(2) biosynthesis and CT26 cell proliferation. The anti-proliferative effects of COX inhibition are rescued specifically by treatment with PGE(2) or the EP4 receptor-selective agonist PGE(1)-OH via phosphatidylinositol 3-kinase/extracellular signal-regulated kinase (ERK) activation, thus providing a functional link between PGE(2)-induced cell proliferation and EP4-mediated ERK signaling. Indomethacin or COX2 inhibitors, but not COX1 inhibitors, reduced the size and number of CT26-derived tumors in vivo. These inhibitory effects are paralleled by marked declines in the levels of tumor PGE(2), suggesting that their anti-tumor effects are directly associated with the inhibition of COX2 enzymatic activity. The described anti-tumor effects of indomethacin are evident whether it is administered at the time of, or 7 days after, tumor cell injection, suggesting that it has tumor preventive and therapeutic actions. Furthermore, the observation that indomethacin increases the survival rates of tumor-bearing mice, even after withdrawal of the drug, indicates that its effects are long lasting and that it may be potentially useful for the prevention and the clinical management of human cancers. PMID- 15123664 TI - Degradation of pulmonary surfactant protein D by Pseudomonas aeruginosa elastase abrogates innate immune function. AB - The alveolar epithelium is lined by surfactant, a lipoprotein complex that both reduces surface tension and mediates several innate immune functions including bacterial aggregation, alteration of alveolar macrophage function, and regulation of bacterial clearance. Surfactant protein-D (SP-D) participates in several of these immune functions, and specifically it enhances the clearance of the pulmonary pathogen Pseudomonas aeruginosa, a common cause of morbidity and mortality in cystic fibrosis (CF) patients. P. aeruginosa secretes a variety of virulence factors including elastase, a zinc-metalloprotease, which degrades both SP-A and SP-D. Here we show that SP-D is cleaved by elastase to produce a stable 35-kDa fragment in a time-, temperature-, and dose-dependent manner. Degradation is inhibited by divalent metal cations, a metal chelator, and the elastase inhibitor, phosphoramidon. Sequencing the SP-D degradation products localized the major cleavage sites to the C-terminal lectin domain. The SP-D fragment fails to bind or aggregate bacteria that are aggregated by intact SP-D. SP-D fragment is observed when normal rat bronchoalveolar lavage (BAL) is treated with Pseudomonas aeruginosa elastase, and SP-D fragments are present in the BAL of CF lung allograft patients. These data show that degradation of SP-D occurs in the BAL environment and that degradation eliminates many normal immune functions of SP-D. PMID- 15123665 TI - Peptide inhibition of catalytic and noncatalytic activities of matrix metalloproteinase-9 blocks tumor cell migration and invasion. AB - Migration of invasive cells appears to be dependent on matrix metalloproteinases (MMPs) anchored on the cell surface through integrins. We have previously demonstrated an interaction between the integrin alpha-subunit I domain and the catalytic domain of MMP-9. We now show that there is also an interaction between the integrin beta subunit and MMP-9. Using phage display, we have developed MMP-9 inhibitors that bind either to the MMP-9 catalytic domain, the collagen binding domain, or the C-terminal hemopexin-like domain. The C-terminal domain-binding peptide mimics an activation epitope in the stalk of the integrin beta chain and inhibits the association of MMP-9 C-terminal domain with alpha(V)beta(5) integrin. Unlike other MMP-9 binding peptides, it does not directly inhibit catalytic activity of MMP-9, but still prevents proenzyme activation and cell migration in vitro and tumor xenograft growth in vivo. We also find an association between MMP-9 and urokinase-plasminogen activator receptor and find that urokinase-plasminogen activator receptor is cleaved by MMP-9. Collectively, we have defined molecular details for several interactions mediated by the different MMP-9 domains. PMID- 15123666 TI - The role of protein kinase D in neurotensin secretion mediated by protein kinase C-alpha/-delta and Rho/Rho kinase. AB - Neurotensin (NT) is a gut peptide that plays an important role in gastrointestinal (GI) secretion, motility, and growth as well as the proliferation of NT receptor positive cancers. Secretion of NT is regulated by phorbol ester-sensitive protein kinase C (PKC) isoforms-alpha and -delta and may involve protein kinase D (PKD). The purpose of our present study was: (i) to define the role of PKD in NT release from BON endocrine cells and (ii) to delineate the upstream signaling mechanisms mediating this effect. Here, we demonstrate that small interfering RNA (siRNA) targeted against PKD dramatically inhibited both basal and PMA-stimulated NT secretion; NT release is significantly increased by overexpression of PKD. PKC-alpha and -delta siRNA attenuated PKD activity, whereas overexpression of PKC-alpha and -delta enhanced PKD activity. Rho kinase (ROK) siRNA significantly inhibited NT secretion, whereas overexpression of ROKalpha effectively increased NT release. Rho protein inhibitor C3 dramatically inhibited both NT secretion and PKD activity. In conclusion, our results demonstrate that PKD activation plays a central role in NT peptide secretion; upstream regulators of PKD include PKC-alpha and -delta and Rho/ROK. Importantly, our results identify novel signaling pathways, which culminate in gut peptide release. PMID- 15123667 TI - The N-glycosidase activity of the ribosome-inactivating protein ME1 targets single-stranded regions of nucleic acids independent of sequence or structural motifs. AB - ME(1), a type I ribosome-inactivating protein (RIP), belongs to a family of enzymes long believed to possess rRNA N-glycosidase activity directed solely at the universally conserved residue A4324 in the sarcin/ricin loop of large eukaryotic and prokaryotic rRNAs. We have investigated the effect of modifying the structure of nonribosomal RNA substrates on their interaction with ME(1) and other RIPs. ME(1) was shown to depurinate a variety of partially denatured nucleic acids, randomly removing adenine residues from single-stranded regions and, to a lesser extent, guanine residues from wobble base-pairs in hairpin stems. A defined sequence motif was not required for recognition of non-paired adenosines and cleavage of the N-glycosidic bond. Substrate recognition and ME(1) activity appeared to depend on the physical availability of nucleotides, and denaturation of nucleic acid substrates increased their interaction with ME(1). Pretreatment of mRNA at 75 degrees C rather than 60 degrees C, for example, lowered the apparent K(D) from 87.1 to 73.9 nm, making it more vulnerable to depurination by RIPs. Exposure to ME(1) in vitro completely abolished the infectivity of partially denatured RNA transcripts of the potato spindle tuber viroid, suggesting that RIPs may target invading nucleic acids before they reach host ribosomes in vivo. Our data suggest that the extensive folding of many potential substrates interferes with their ability to interact with RIPs, thereby blocking their inactivation by ME(1) (or other RIPs). PMID- 15123668 TI - Voltage dependence of H+ buffering mediated by sodium bicarbonate cotransport expressed in Xenopus oocytes. AB - The electrogenic sodium bicarbonate cotransporter (NBCe1) is expressed in many epithelial cells and, in the brain, in glial cells. Little is known about the physiological significance of the NBCe1 for proton homeostasis and for other acid/base-coupled transporters in these cells. We have measured the voltage dependent transport activity of an NBC from human kidney, type hkNBCe1, expressed in oocytes of the frog Xenopus laevis, by recording membrane current and the changes in intracellular pH and sodium at different membrane potentials between 20 and -100 mV. The apparent intracellular buffer capacity was increased and became dependent upon membrane voltage when the NBCe1 was expressed; the measured buffer capacity increased by up to 7 mm/10 mV of membrane depolarization. Lactate transport by the electroneutral monocarboxylate transporter became enhanced and dependent upon membrane potential, when the monocarboxylate transporter (isoform 1) was co-expressed with NBCe1 in oocytes. Our results indicate that the electrogenic NBCe1 renders the cell membrane potential an effective regulator of intracellular H(+) buffering and acid/base-coupled metabolite transport. PMID- 15123669 TI - Regulation of neuronal voltage-gated sodium channels by the ubiquitin-protein ligases Nedd4 and Nedd4-2. AB - Nedd4 and Nedd4-2 are ubiquitin-protein ligases known to regulate a number of membrane proteins including receptors and ion transporters. Regulation of the epithelial Na(+) channel by Nedd4 and Nedd4-2 is mediated via interactions between the PY motifs of the epithelial sodium channel subunits and the Nedd4/Nedd4-2 WW domains. This example serves as a model for the regulation of other PY motif-containing ion channels by Nedd4 and Nedd4-2. We found that the carboxyl termini of the six voltage-gated Na(+) (Na(v)) channels contain typical PY motifs (PPXY), and a further Na(v) contains a PY motif variant (LPXY). Not only did we demonstrate by Far-Western analysis that Nedd4 and Nedd4-2 interact with the PY motif-containing Na(v) channels, but we also showed that these channels have conserved WW domain binding specificity. We further showed that the carboxyl termini fusion proteins of one central nervous system and one peripheral nervous system-derived Na(+) channel (Na(v)1.2 and Na(v)1.7, respectively) are readily ubiquitinated by Nedd4-2. In Xenopus oocytes, Nedd4-2 strongly inhibited the activities of all three Na(v)s (Na(v)1.2, Na(v)1.7, and Na(v)1.8) tested. Interestingly, Nedd4 suppressed the activity of Na(v)1.2 and Na(v)1.7 but was a poor inhibitor of Na(v)1.8. Our results provide evidence that Nedd4 and Nedd4-2 are likely to be key regulators of specific neuronal Na(v) channels in vivo. PMID- 15123670 TI - Functional expression of TRESK-2, a new member of the tandem-pore K+ channel family. AB - A new member of the tandem-pore K+ (K(2P)) channel family has been isolated from mouse testis complementary DNA. The new K(2P) channel was named TRESK-2, as its amino acid sequence shares 65% identity with that of TRESK-1. Mouse TRESK-2 is a 394-amino acid protein and possesses four putative transmembrane segments and two pore-forming domains. TRESK-2 has a long cytoplasmic domain joining the second and third transmembrane segments and a short carboxyl terminus. In the rat, TRESK 2 mRNA transcripts were expressed abundantly in the thymus and spleen and at low levels in many other tissues, including heart, small intestine, skeletal muscle, uterus, testis, and placenta, as judged by Northern blot analysis. TRESK-2 mRNA was also expressed in mouse and human tissues. In COS-7 cells transfected with TRESK-2 DNA, a time-independent and noninactivating K+-selective current was recorded. TRESK-2 was insensitive to 1 mm tetraethylammonium, 100 nm apamin, 1 mm 4-aminopyridine, and 10 microm glybenclamide. TRESK-2 was inhibited by 10 microm quinidine, 20 microm arachidonate and acid (pH 6.3) at 49, 43, and 23%, respectively. Single channel openings of TRESK-2 showed marked open channel noise. In symmetrical 150 mm KCl, the current-voltage relationship of TRESK-2 was slightly inwardly rectifying, with the single channel conductance 13 picosiemens (pS) at +60 mV and 16 pS at -60 mV. In inside-out patches, TRESK-2 was unaffected by the intracellular application of 10 microm guanosine 5'-O-(thiotriphosphate). These results show that TRESK-2 is a functional member of the K(2P) channel family and contributes to the background K+ conductance in many types of cells. PMID- 15123671 TI - AML1/Runx1 recruits calcineurin to regulate granulocyte macrophage colony stimulating factor by Ets1 activation. AB - Acute myeloid leukemia 1 (AML1), also denoted Runx1, is a transcription factor essential for hematopoiesis, and the AML1 gene is the most common target of chromosomal translocations in human leukemias. AML1 binds to sequences present in the regulatory regions of a number of hematopoiesis-specific genes, including certain cytokines such as granulocyte macrophage colony-stimulating factor (GM CSF) up-regulated after T cell receptor stimulation. Here we show that both subunits of the Ca(2+)/calmodulin-dependent protein phosphatase calcineurin (CN), which is activated upon T cell receptor stimulation, interact directly with the N terminal runt homology domain-containing part of AML1. The regulatory CN subunit binds AML1 with a higher affinity and in addition also interacts with the isolated runt homology domain. The related Runx2 transcription factor, which is essential for bone formation, also interacts with CN. A constitutively active derivative of CN is shown to activate synergistically the GM-CSF promoter/enhancer together with AML1 or Runx2. We also provide evidence that relief of the negative effect of the AML1 sites is important for Ca(2+) activation of the GM-CSF promoter/enhancer and that AML1 overexpression increases this Ca(2+) activation. Both subunits of CN interact with AML1 in coimmunoprecipitation analyses, and confocal microscopy analysis of cells expressing fluorescence-tagged protein derivatives shows that CN can be recruited to the nucleus by AML1 in vivo. Mutant analysis of the GM-CSF promoter shows that the Ets1 binding site of the promoter is essential for the synergy between AML1 and CN in Jurkat T cells. Analysis of the effects of inhibitors of the protein kinase glycogen synthase kinase-3beta and in vitro phosphorylation/dephosphorylation analysis of Ets1 suggest that glycogen synthase kinase-3beta-phosphorylated Ets1 is a target of AML1-recruited CN phosphatase at the GM-CSF promoter. PMID- 15123672 TI - Gbetagamma-activated inwardly rectifying K(+) (GIRK) channel activation kinetics via Galphai and Galphao-coupled receptors are determined by Galpha-specific interdomain interactions that affect GDP release rates. AB - Gbetagamma-activated inwardly rectifying K(+) (GIRK) channels have distinct gating properties when activated by receptors coupled specifically to Galpha(o) versus Galpha(i) subunit isoforms, with Galpha(o)-coupled currents having approximately 3-fold faster agonist-evoked activation kinetics. To identify the molecular determinants in Galpha subunits mediating these kinetic differences, chimeras were constructed using pertussis toxin (PTX)-insensitive Galpha(oA) and Galpha(i2) mutant subunits (Galpha(oA(C351G)) and Galpha(i2(C352G))) and examined in PTX-treated Xenopus oocytes expressing muscarinic m2 receptors and Kir3.1/3.2a channels. These experiments revealed that the alpha-helical N-terminal region (amino acids 1-161) and the switch regions of Galpha(i2) (amino acids 162-262) both partially contribute to slowing the GIRK activation time course when compared with the Galpha(oA(C351G))-coupled response. When present together, they fully reproduce Galpha(i2(C352G))-coupled GIRK kinetics. The Galpha(i2) C terminal region (amino acids 263-355) had no significant effect on GIRK kinetics. Complementary responses were observed with chimeras substituting the Galpha(o) switch regions into the Galpha(i2(C352G)) subunit, which partially accelerated the GIRK activation rate. The Galpha(oA)/Galpha(i2) chimera results led us to examine an interaction between the alpha-helical domain and the Ras-like domain previously implicated in mediating a 4-fold slower in vitro basal GDP release rate in Galpha(i1) compared with Galpha(o). Mutations disrupting the interdomain contact in Galpha(i2(C352G)) at either the alphaD-alphaE loop (R145A) or the switch III loop (L233Q/A236H/E240T/M241T), significantly accelerated the GIRK activation kinetics consistent with the Galpha(i2) interdomain interface regulating receptor-catalyzed GDP release rates in vivo. We propose that differences in Galpha(i) versus Galpha(o)-coupled GIRK activation kinetics are due to intrinsic differences in receptor-catalyzed GDP release that rate-limit Gbetagamma production and is attributed to heterogeneity in Galpha(i) and Galpha(o) interdomain contacts. PMID- 15123673 TI - Complex interplay among regulators of drug resistance genes in Saccharomyces cerevisiae. AB - The Gal4p family of yeast zinc cluster proteins comprises regulators of multidrug resistance genes. For example, Pdr1p and Pdr3p bind as homo- or heterodimers to pleiotropic drug response elements (PDREs) found in promoters of target genes. Other zinc cluster activators of multidrug resistance genes include Stb5p and Yrr1p. To better understand the interplay among these activators, we have performed native co-immunoprecipitation experiments using strains expressing tagged zinc cluster proteins from their natural chromosomal locations. Interestingly, Stb5p is found predominantly as a Pdr1p heterodimer and shows little homodimerization. No interactions of Stb5p with Pdr3p or Yrr1p could be detected in our assays. In contrast to Stb5p, Yrr1p is only detected as a homodimer. Similar results were obtained using glutathione S-transferase pull down assays. Importantly, the purified DNA binding domains of Stb5p and Pdr1p bound to a PDRE as heterodimers in vitro. These results suggest that the DNA binding domains of Pdr1p and Stb5p are sufficient for heterodimerization. Our data demonstrate a complex interplay among these activators and suggest that Pdr1p is a master drug regulator involved in recruiting other zinc cluster proteins to fine tune the regulation of multidrug resistance genes. PMID- 15123674 TI - Structural basis for coronavirus-mediated membrane fusion. Crystal structure of mouse hepatitis virus spike protein fusion core. AB - The surface transmembrane glycoprotein is responsible for mediating virion attachment to cell and subsequent virus-cell membrane fusion. However, the molecular mechanisms for the viral entry of coronaviruses remain poorly understood. The crystal structure of the fusion core of mouse hepatitis virus S protein, which represents the first fusion core structure of any coronavirus, reveals a central hydrophobic coiled coil trimer surrounded by three helices in an oblique, antiparallel manner. This structure shares significant similarity with both the low pH-induced conformation of influenza hemagglutinin and fusion core of HIV gp41, indicating that the structure represents a fusion-active state formed after several conformational changes. Our results also indicate that the mechanisms for the viral fusion of coronaviruses are similar to those of influenza virus and HIV. The coiled coil structure has unique features, which are different from other viral fusion cores. Highly conserved heptad repeat 1 (HR1) and HR2 regions in coronavirus spike proteins indicate a similar three dimensional structure among these fusion cores and common mechanisms for the viral fusion. We have proposed the binding regions of HR1 and HR2 of other coronaviruses and a structure model of their fusion core based on our mouse hepatitis virus fusion core structure and sequence alignment. Drug discovery strategies aimed at inhibiting viral entry by blocking hairpin formation may be applied to the inhibition of a number of emerging infectious diseases, including severe acute respiratory syndrome. PMID- 15123675 TI - Mutational analysis of the archaeal tyrosine recombinase SSV1 integrase suggests a mechanism of DNA cleavage in trans. AB - The only tyrosine recombinase so far studied in archaea, the SSV1 integrase, harbors several changes in the canonical residues forming the catalytic pocket of this family of recombinases. This raised the possibility of a different mechanism for archaeal tyrosine recombinase. The residues of Int(SSV) tentatively involved in catalysis were modified by site-directed mutagenesis, and the properties of the corresponding mutants were studied. The results show that all of the targeted residues are important for activity, suggesting that the archaeal integrase uses a mechanism similar to that of bacterial or eukaryotic tyrosine recombinases. In addition, we show that Int(SSV) exhibits a type IB topoisomerase activity because it is able to relax both positive and negative supercoils. Interestingly, in vitro complementation experiments between the inactive integrase mutant Y314F and all other inactive mutants restore in all cases enzymatic activity. This suggests that, as for the yeast Flp recombinase, the active site is assembled by the interaction of the tyrosine from one monomer with the other residues from another monomer. The shared active site paradigm of the eukaryotic Flp protein may therefore be extended to the archaeal tyrosine recombinase Int(SSV). PMID- 15123676 TI - Allosteric beta1 integrin antibodies that stabilize the low affinity state by preventing the swing-out of the hybrid domain. AB - The ligand binding function of integrins can be modulated by various monoclonal antibodies by both direct and indirect mechanisms. We have characterized an anti beta(1) antibody, SG/19, that had been reported to inhibit the function of the beta(1) integrin on the cell surface. SG/19 recognized the wild type beta(1) subunit that exists in a conformational equilibrium between the high and low affinity states but bound poorly to a mutant beta(1) integrin that had been locked in a high affinity state. Epitope mapping of SG/19 revealed that Thr(82) in the beta(1) subunit, located at the outer face of the boundary between the I like and hybrid domains, was the key binding determinant for this antibody. Direct visualization of the alpha (5)beta(1) headpiece fragment in complex with SG/19 Fab with electron microscopy confirmed the location of the binding surface and showed that the ligand binding site is not occluded by the bound Fab. Surface plasmon resonance showed that alpha (5)beta(1) integrin bound by SG/19 maintained a low affinity toward its physiological ligand fibronectin (Fn) whereas binding by function-blocking anti-alpha(5) antibodies resulted in a complete loss of fibronectin binding. Thus a class of the anti-beta antibodies represented by SG/19 attenuate the ligand binding function by restricting the conformational shift to the high affinity state involving the swing-out of the hybrid domain without directly interfering with ligand docking. PMID- 15123677 TI - Differential effects of the SR proteins 9G8, SC35, ASF/SF2, and SRp40 on the utilization of the A1 to A5 splicing sites of HIV-1 RNA. AB - Splicing is a crucial step for human immunodeficiency virus, type 1 (HIV-1) multiplication; eight acceptor sites are used in competition to produce the vif, vpu, vpr, nef, env, tat, and rev mRNAs. The effects of SR proteins have only been investigated on a limited number of HIV-1 splicing sites by using small HIV-1 RNA pieces. To understand how SR proteins influence the use of HIV-1 splicing sites, we tested the effects of overproduction of individual SR proteins in HeLa cells on the splicing pattern of an HIV-1 RNA that contained all the splicing sites. The steady state levels of the HIV-1 mRNAs produced were quantified by reverse transcriptase-PCR. For interpretation of the data, transcripts containing one or several of the HIV-1 acceptor sites were spliced in vitro in the presence or the absence of one of the tested SR proteins. Both in vivo and in vitro, acceptor sites A2 and A3 were found to be strongly and specifically regulated by SR proteins. ASF/SF2 strongly activates site A2 and to a lesser extent site A1. As a result, upon ASF/SF2 overexpression, the vpr mRNA steady state level is specifically increased. SC35 and SRp40, but not 9G8, strongly activate site A3, and their overexpression ex vivo induces a dramatic accumulation of the tat mRNA, to the detriment of most of the other viral mRNAs. Here we showed by Western blot analysis that the Nef protein synthesis is strongly decreased by overexpression of SC35, SRp40, and ASF/SF2. Finally, activation by ASF/SF2 and 9G8 was found to be independent of the RS domain. This is the first investigation of the effects of variations of individual SR protein concentrations that is performed ex vivo on an RNA containing a complex set of splicing sites. PMID- 15123678 TI - Insulin induces SOCS-6 expression and its binding to the p85 monomer of phosphoinositide 3-kinase, resulting in improvement in glucose metabolism. AB - The suppressors of cytokine signaling (SOCS) family is thought to act largely as a negative regulator of signaling by cytokines and some growth factors. Surprisingly, the SOCS-6 transgenics had no significant defects in the cytokine signaling and hematopoietic system but displayed significant improvements in glucose metabolism. Insulin stimulation of Akt/protein kinase B was also potentiated. Biochemical analysis showed that, after insulin stimulation, SOCS-6 interacted with the monomeric p85 subunit of class-Ia phosphoinositide (PI) 3 kinase but not with p85/p110 dimers. Furthermore, SOCS-6 expression is transiently increased by serum and insulin in normal fibroblasts. However, both the mRNA and protein of SOCS-6 were rapidly degraded after induction by insulin. The degradation of the SOCS-6 protein was partially inhibited by a proteasome inhibitor, suggesting a proteasome-mediated degradation mechanism. In contrast, SOCS-6-associated p85 was not degraded and could be recruited to the newly synthesized SOCS-6 molecules in the presence of insulin, suggesting that SOCS-6 expression and its interaction with p85, but not the degradation, is regulated by insulin. The phenotype of SOCS-6 transgenic mice bears a striking resemblance to p85 knock-out mouse models in which glucose metabolism stimulated by insulin is significantly improved despite reduced activation of PI 3-kinase. This suggests that monomeric p85 might play a physiologically important role in attenuating signaling through PI 3-kinase-dependent pathways in unstimulated cells. Therefore, our results indicate that SOCS-6 may provide a dynamically regulated mechanism by which insulin can transiently overcome the negative effects that p85 monomers have on signaling via PI 3-kinase-dependent signaling pathways. PMID- 15123679 TI - An Arg307 to Gln polymorphism within the ATP-binding site causes loss of function of the human P2X7 receptor. AB - The P2X(7) receptor is a ligand-gated channel that is highly expressed on mononuclear cells of the immune system and that mediates ATP-induced apoptosis. Wide variations in the function of the P2X receptor have been observed, explained in part by (7)loss-of-function polymorphisms that change Glu(496) to Ala (E496A) and Ile(568) to Asn (I568N). In this study, a third polymorphism, which substitutes an uncharged glutamine for the highly positively charged Arg(307) (R307Q), has been found in heterozygous dosage in 12 of 420 subjects studied. P2X(7) function was measured by ATP-induced fluxes of Rb(+), Ba(2+), and ethidium(+) into peripheral blood monocytes or various lymphocyte subsets and was either absent or markedly decreased. Transfection experiments showed that P2X(7) carrying the R307Q mutation lacked either channel or pore function despite robust protein synthesis and surface expression of the receptor. The monoclonal antibody (clone L4) that binds to the extracellular domain of wild type P2X(7) and blocks P2X(7) function failed to bind to the R307Q mutant receptor. Differentiation of monocytes to macrophages up-regulated P2X(7) function in cells heterozygous for the R307Q to a value 10-40% of that for wild type macrophages. However, macrophages from a subject who was double heterozygous for R307Q/I568N remained totally non-functional for P2X(7), and lymphocytes from the same subject also lacked ATP-stimulated phospholipase D activity. These data identify a third loss of-function polymorphism affecting the human P2X(7) receptor, and since the affected Arg(307) is homologous to those amino acids essential for ATP binding to P2X(1) and P2X(2), it is likely that this polymorphism abolishes the binding of ATP to the extracellular domain of P2X(7). PMID- 15123681 TI - Insulin receptor substrate-2-dependent interleukin-4 signaling in macrophages is impaired in two models of type 2 diabetes mellitus. AB - We have shown previously that hyperinsulinemia inhibits interferon-alpha dependent activation of phosphatidylinositol 3-kinase (PI3-kinase) through mammalian target of rapamycin (mTOR)-induced serine phosphorylation of insulin receptor substrate (IRS)-1. Here we report that chronic insulin and high glucose synergistically inhibit interleukin (IL)-4-dependent activation of PI3-kinase in macrophages via the mTOR pathway. Resident peritoneal macrophages (PerMPhis) from diabetic (db/db) mice showed a 44% reduction in IRS-2-associated PI3-kinase activity stimulated by IL-4 compared with PerMPhis from heterozygote (db/+) control mice. IRS-2 from db/db mouse PerMPhis also showed a 78% increase in Ser/Thr-Pro motif phosphorylation without a difference in IRS-2 mass. To investigate the mechanism of this PI3-kinase inhibition, 12-O tetradecanoylphorbol-13-acetate-matured U937 cells were treated chronically with insulin (1 nm, 18 h) and high glucose (4.5 g/liter, 48 h). In these cells, IL-4 stimulated IRS-2-associated PI3-kinase activity was reduced by 37.5%. Importantly, chronic insulin or high glucose alone did not impact IL-4-activated IRS-2-associated PI3-kinase. Chronic insulin + high glucose did reduce IL-4 dependent IRS-2 tyrosine phosphorylation and p85 association by 54 and 37%, respectively, but did not effect IL-4-activated JAK/STAT signaling. When IRS-2 Ser/Thr-Pro motif phosphorylation was examined, chronic insulin + high glucose resulted in a 92% increase in IRS-2 Ser/Thr-Pro motif phosphorylation without a change in IRS-2 mass. Pretreatment of matured U937 cells with rapamycin blocked chronic insulin + high glucose-dependent IRS-2 Ser/Thr-Pro motif phosphorylation and restored IL-4-dependent IRS-2-associated PI3-kinase activity. Taken together these results indicate that IRS-2-dependent IL-4 signaling in macrophages is impaired in models of type 2 diabetes mellitus through a mechanism that relies on insulin/glucose-dependent Ser/Thr-Pro motif serine phosphorylation mediated by the mTOR pathway. PMID- 15123680 TI - The peroxisome proliferator-activated receptor alpha (PPARalpha) agonist ciprofibrate inhibits apolipoprotein B mRNA editing in low density lipoprotein receptor-deficient mice: effects on plasma lipoproteins and the development of atherosclerotic lesions. AB - Low density lipoprotein receptor (LDLR)-deficient mice fed a chow diet have a mild hypercholesterolemia caused by the abnormal accumulation in the plasma of apolipoprotein B (apoB)-100- and apoB-48-carrying intermediate density lipoproteins (IDL) and low density lipoproteins (LDL). Treatment of LDLR deficient mice with ciprofibrate caused a marked decrease in plasma apoB-48 carrying IDL and LDL but at the same time caused a large accumulation of triglyceride-depleted apoB-100-carrying IDL and LDL, resulting in a significant increase in plasma cholesterol levels. These plasma lipoprotein changes were associated with an increase in the hepatic secretion of apoB-100-carrying very low density lipoproteins (VLDL) and a decrease in the secretion of apoB-48 carrying VLDL, accompanied by a significant decrease in hepatic apoB mRNA editing. Hepatic apobec-1 complementation factor mRNA and protein abundance were significantly decreased, whereas apobec-1 mRNA and protein abundance remained unchanged. No changes in apoB mRNA editing occurred in the intestine of the treated animals. After 150 days of treatment with ciprofibrate, consistent with the increased plasma accumulation of apoB-100-carrying IDL and LDL, the LDLR deficient mice displayed severe atherosclerotic lesions in the aorta. These findings demonstrate that ciprofibrate treatment decreases hepatic apoB mRNA editing and alters the pattern of hepatic lipoprotein secretion toward apoB-100 associated VLDL, changes that in turn lead to increased atherosclerosis. PMID- 15123682 TI - The residue 129 polymorphism in human prion protein does not confer susceptibility to Creutzfeldt-Jakob disease by altering the structure or global stability of PrPC. AB - There are two common forms of prion protein (PrP) in humans, with either methionine or valine at position 129. This polymorphism is a powerful determinant of the genetic susceptibility of humans toward both sporadic and acquired forms of prion disease and restricts propagation of particular prion strains. Despite its key role, we have no information on the effect of this mutation on the structure, stability, folding, and dynamics of the cellular form of PrP (PrP(C)). Here, we show that the mutation has no measurable effect on the folding, dynamics, and stability of PrP(C). Our data indicate that the 129M/V polymorphism does not affect prion propagation through its effect on PrP(C); rather, its influence is likely to be downstream in the disease mechanism. We infer that the M/V effect is mediated through the conformation or stability of disease-related PrP (PrP(Sc)) or intermediates or on the kinetics of their formation. PMID- 15123683 TI - Frataxin-mediated iron delivery to ferrochelatase in the final step of heme biosynthesis. AB - Human ferrochelatase, a mitochondrial membrane-associated protein, catalyzes the terminal step of heme biosynthesis by insertion of ferrous iron into protoporphyrin IX. The recently solved x-ray structure of human ferrochelatase identifies a potential binding site for an iron donor protein on the matrix side of the homodimer. Herein we demonstrate Hs holofrataxin to be a high affinity iron binding partner for Hs ferrochelatase that is capable of both delivering iron to ferrochelatase and mediating the terminal step in mitochondrial heme biosynthesis. A general regulatory mechanism for mitochondrial iron metabolism is described that defines frataxin involvement in both heme and iron-sulfur cluster biosyntheses. In essence, the distinct binding affinities of holofrataxin to the target proteins, ferrochelatase (heme synthesis) and ISU (iron-sulfur cluster synthesis), allows discrimination between the two major iron-dependent pathways and facilitates targeted heme biosynthesis following down-regulation of frataxin. PMID- 15123684 TI - Functional mapping of Ca2+ signaling complexes in plasma membrane microdomains of polarized cells. AB - Many cells cluster signaling complexes in plasma membrane microdomains. Polarized secretory cells cluster all Ca2+ signaling proteins, including GPCRs, at the apical pole. The functional significance of such an arrangement is not known because of a lack of techniques for functional mapping of signaling complexes at plasma membrane patches. In the present work, we developed such a technique based on the use of two patch pipettes, a recording and a stimulating pipette (SP). Including 20% glycerol in the SP solution increased the viscosity and the hydrophobicity to prevent leakage and formation of tight seals on the plasma membrane. This allowed moving the SP between sites to stimulate multiple patches of the same cell and with the same agonist concentrations. Functional mapping of Ca2+ signaling in pancreatic acinar cells revealed that the M3, cholecystokinin, and bombesin signaling complexes at the apical pole are much more sensitive to stimulation than those at the basal pole. Furthermore, at physiological agonist concentrations, Ca2+ signals could be evoked only by stimulation of membrane patches at the apical pole. [Ca2+](i) imaging revealed that Ca2+ waves were invariably initiated at the site of apical membrane patch stimulation, suggesting that long range diffusion of second messengers is not obligatory to initiate and propagate apical-to-basal Ca2+ waves. The present studies reveal a remarkable heterogeneity in responsiveness of Ca2+ signaling complexes at membrane microdomains, with the most responsive complexes confined to the apical pole, probably to restrict the Ca2+ signals to the site of exocytosis and allow the polarized functions of secretory cells. PMID- 15123685 TI - Conditional expression of 15-lipoxygenase-1 inhibits the selenoenzyme thioredoxin reductase: modulation of selenoproteins by lipoxygenase enzymes. AB - The selenoenzyme thioredoxin reductase regulates redox-sensitive proteins involved in inflammation and carcinogenesis, including ribonucleotide reductase, p53, NFkappaB, and others. Little is known about endogenous cellular factors that modulate thioredoxin reductase activity. Here we report that several metabolites of 15-lipoxygenase-1 inhibit purified thioredoxin reductase in vitro. 15(S) Hydroperoxy-5,8,11-cis-13-trans-eicosatetraenoic acid, a metastable hydroperoxide generated by 15-lipoxygenase-1, and 4-hydroxy-2-nonenal, its non-enzymatic rearrangement product inhibit thioredoxin reductase with IC(50) = 13 +/- 1.5 microm and 1 +/- 0.2 microm, respectively. Endogenously generated metabolites of 15-lipoxygenase-1 also inhibit thioredoxin reductase in HEK-293 cells that harbor a 15-LOX-1 gene under the control of an inducible promoter complex. Conditional, highly selective induction of 15-lipoxygenase-1 caused an inhibition of ribonucleotide reductase activity, cell cycle arrest in G(1), impairment of anchorage-independent growth, and accumulation of the pro-apoptotic protein BAX. All of these responses are consistent with inhibition of thioredoxin reductase via 15-lipoxygenase-1 overexpression. In contrast, metabolites of 5-lipoxygenase were poor inhibitors of isolated thioredoxin reductase, and the overexpression of 5-lipoxygenase did not inhibit thioredoxin reductase or cause a G cell cycle arrest. The influences of 15-lipoxygenase-1 on (1)inflammation, cell growth, and survival may be attributable, in part, to inhibition of thioredoxin reductase and several redox-sensitive processes subordinate to thioredoxin reductase. PMID- 15123686 TI - An activin mutant with disrupted ALK4 binding blocks signaling via type II receptors. AB - Activins control many physiologic and pathophysiologic processes in multiple tissues and, like other TGF-beta superfamily members, signal via type II (ActRII/IIB) and type I (ALK4) receptor serine kinases. ActRII/IIB are promiscuous receptors known to bind at least a dozen TGF-beta superfamily ligands including activins, myostatin, several BMPs, and nodal. Here we utilize a new screening procedure to rapidly identify activin-A mutants with loss of signaling activity. Our goal was to identify activin-A mutants able to bind ActRII but unable to bind ALK4 and which would be, therefore, candidate type II activin receptor antagonists. Using the structure of BMP-2 bound to its type I receptor (ALK3) as a guide, we introduced mutations in the context of the inhibin betaA cDNA and assessed the signaling activity of the resulting mutant proteins. We identified several mutants in the finger (M91E, I105E, M108A) and wrist (activin A/activin C chimera, S60P, I63P) regions of activin-A with reduced signaling activity. Of these the M108A mutant displayed the lowest signaling activity while retaining wild-type-like affinity for ActRII. Unlike wild-type activin-A, the M108A mutant was unable to form a cross-linked complex with ALK4 in the presence of ActRII indicating that its ability to bind ALK4 was disrupted. This data suggested that the M108A mutant might be capable of modulating signaling of activin and related ligands. Indeed, the M108A mutant antagonized activin-A and myostatin, but not TGF-beta, signaling in 293T cells, indicating it may be generally capable of blocking ligands that signal via ActRII/IIB. PMID- 15123687 TI - The androgen receptor acetylation site regulates cAMP and AKT but not ERK-induced activity. AB - The androgen receptor (AR) regulates ligand-dependent gene transcription upon binding specific DNA sequences. The AR conveys both trans-activation and trans repression functions, which together contribute to prostate cellular growth, differentiation, and apoptosis. Like histone H3, the AR is post-translationally modified by both acetylation and phosphorylation. The histone acetyltransferase p300 transactivates the AR and directly acetylates the AR in vitro at a conserved motif. Point mutations of the AR acetylation motif that abrogate acetylation reduce trans-activation by p300 without affecting the trans-repression function of the AR. The current studies assessed the functional relationship between acetylation and phosphorylation of the AR. Herein trans-activation of the AR acetylation site mutants were enhanced by the p42/p44 MAPK pathway but were defective in regulation by protein kinase A (PKA) signaling. PKA inhibition augmented ARwt activity but not AR acetylation mutant gene reporter activity and association at an androgen response element in chromatin immunoprecipitation assays. Mutations of the lysine residues at the AR acetylation site reduced trichostatin A (TSA) responsiveness and ligand-induced phosphorylation of the AR. The AR acetylation site mutant formed ligand-induced phosphorylation-dependent isoforms with distinguishable characteristics from wild type AR as determined with two-dimensional electrophoresis. Conversely, point mutation of a subset of AR phosphorylation sites reduced trichostatin A responsiveness and trans activation by histone acetyltransferases. Together these studies suggest that acetylation and phosphorylation of the AR are linked events and that the conserved AR lysine motif contributes to a select subset of pathways governing AR activity. PMID- 15123688 TI - Critical role of residues defining the ligand binding pocket in hepatocyte nuclear factor-4alpha. AB - Hepatocyte nuclear factor-4alpha (HNF-4alpha), a member of the nuclear receptor superfamily, is a crucial regulator of a large number of genes involved in glucose, cholesterol, and fatty acid metabolism. Unlike other members of the superfamily, HNF-4alpha activates transcription in the absence of exogenously added ligand. Recently published crystallographic data show that fatty acids are endogenous ligands for HNF-4. Transcriptional analysis of point mutations of the residues that are located in helices H3, H5, H10, and H11, which have been shown to come in contact with the ligand, resulted in a dramatic decrease in activity, without affecting DNA binding and dimerization. Our results show the importance of residues Ser-181, Met-182 in H3, Leu-219, Leu-220 and Arg-226 in H5, Ile-338 in H10, and Ile-346 in H11 that line the ligand-binding domain pocket in HNF 4alpha and impair its transactivation potential. Structural modeling reveals that the mutations do not cause any large scale structural alterations, and the observed loss in transactivation can be attributed to local changes, demonstrating that these residues play a significant role in maintaining the structural integrity of the HNF-4alpha ligand binding pocket. PMID- 15123689 TI - Interleukin-7 inactivates the pro-apoptotic protein Bad promoting T cell survival. AB - Interleukin-7 (IL-7) is a cytokine that is required for T cell development and survival. The anti-apoptotic function of IL-7 is partly through induction of Bcl 2 synthesis and cytosolic retention of Bax. Here we show that the Bcl-2 homology 3 domain-only protein, Bad, is involved in cell death following IL-7 withdrawal from D1 cells, an IL-7-dependent murine thymocyte cell line. IL-7 stimulation resulted in the inactivation of Bad by phosphorylation at Ser-112, -136, and 155. The phosphoinositide 3-kinase (PI3K)/Akt pathway has been implicated previously in Bad phosphorylation. In response to IL-7, the PI3K/Akt pathway induced phosphorylation at Ser-136 and -155, but Ser-112 was partly independent of the PI3K/Akt pathway, indicating an as yet unknown pathway in this response. Following IL-7 withdrawal, dephosphorylated Bad translocated from cytosol to mitochondria, bound to Bcl-2, and accelerated cell death. Thus, the inactivation of Bad contributes to the survival function of IL-7. PMID- 15123691 TI - Sue1p is required for degradation of labile forms of altered cytochromes C in yeast mitochondria. AB - Previous studies on certain altered holo-isocytochromes c revealed a rho(-) dependent degradation (RDD) phenotype, in which certain altered holo-iso-1 cytochromes c are at normal or nearly normal levels in rho+ strains, but are at low levels or absent in rho- strains, although wild-type holo-iso-1-cytochrome c is present at normal levels in both rho+ and related rho- strains. The diminished levels of altered holo-iso-1-cytochrome c are due to the rapid degradation that is carried out by a novel proteolytic pathway in the IMS of mitochondria. SUE1, a nuclear gene that encodes a mitochondrial protein, was identified with a genetic screen for mutants that diminish RDD. The levels of RDD and certain other types of altered holo-iso-1-cytochrome c were elevated in rho- sue1 strains. Also, rho+ sue1 strains containing certain altered holo-iso-1-cytochromes c grew better on non-fermentable carbon sources than the corresponding rho+ SUE1 strains. These results indicate that Sue1p may play an important role in the degradation of abnormal holo-iso-1-cytochrome c in the mitochondria. PMID- 15123690 TI - Sequence-specific interaction between mitochondrial Fe-S scaffold protein Isu and Hsp70 Ssq1 is essential for their in vivo function. AB - Isu, the scaffold for assembly of Fe-S clusters in the yeast mitochondrial matrix, is a substrate protein for the Hsp70 Ssq1 and the J-protein Jac1 in vitro. As expected for an Hsp70-substrate interaction, the formation of a stable complex between Isu and Ssq1 requires Jac1 in the presence of ATP. Here we report that a conserved tripeptide, PVK, of Isu is critical for interaction with Ssq1 because amino acid substitutions in this tripeptide inhibit both the formation of the Isu-Ssq1 complex and the ability of Isu to stimulate the ATPase activity of Ssq1. These biochemical defects correlate well with the growth defects of cells expressing mutant Isu proteins. We conclude that the Ssq1-Isu substrate interaction is critical for Fe-S cluster biogenesis in vivo. The ability of Jac1 and mutant Isu proteins to cooperatively stimulate the ATPase activity of Ssq1 was also measured. Increasing the concentration of Jac1 and mutant Isu together but not individually partially overcame the effect of the reduced affinity of the Isu mutant proteins for Ssq1. These results, along with the observation that overexpression of Jac1 was able to suppress the growth defect of an ISU mutant, support the hypothesis that Isu is "targeted" to Ssq1 by Jac1, with a preformed Jac1-Isu complex interacting with Ssq1. PMID- 15123692 TI - Merlin, a tumor suppressor, interacts with transactivation-responsive RNA-binding protein and inhibits its oncogenic activity. AB - The neurofibromatosis type 2 gene-encoded protein, merlin, is related to the ERM (ezrin, radixin, and moesin) family of membrane-cytoskeleton-associated proteins. Recent studies suggest that the loss of neurofibromatosis type 2 function contributes to tumor development and metastasis. Although the cellular functions of merlin as a tumor suppressor are relatively well characterized, the cellular mechanism whereby merlin controls cell proliferation from membrane locations is still poorly understood. During our efforts to find potential merlin modulators through protein-protein interactions, we identified transactivation-responsive RNA-binding protein (TRBP) as a merlin-binding protein in a yeast two-hybrid screen. The interaction between TRBP and merlin was confirmed by glutathione S transferase pull-down assays, co-immunoprecipitation, and co-localization experiments. The carboxyl-terminal regions of each protein were responsible for their interaction. Cells overexpressing TRBP showed enhanced cell growth in cell proliferation assays and also exhibited transformed phenotypes, such as anchorage independent cell growth and tumor development in mouse xenografts. Merlin efficiently inhibited these oncogenic activities of TRBP in our experiments. These results provide the first clue to the functional interaction between TRBP and merlin and suggest a novel mechanism for the tumor suppressor function of merlin both in vitro and in vivo. PMID- 15123693 TI - Sec22p export from the endoplasmic reticulum is independent of SNARE pairing. AB - Molecularly distinct sets of SNARE proteins localize to specific intracellular compartments and catalyze membrane fusion events. Although their central role in membrane fusion is appreciated, little is known about the mechanisms by which individual SNARE proteins are targeted to specific organelles. Here we investigated functional domains in Sec22p that direct this SNARE protein to the endoplasmic reticulum (ER), to Golgi membranes, and into SNARE complexes with Bet1p, Bos1p, and Sed5p. A series of Sec22p deletion mutants were monitored in COPII budding assays, subcellular fractionation gradients, and SNARE complex immunoprecipitations. We found that the N-terminal "profilin-like" domain of Sec22p was required but not sufficient for COPII-dependent export of Sec22p from the ER. Interestingly, versions of Sec22p that lacked the N-terminal domain were assembled into ER/Golgi SNARE complexes. Analyses of Sec22p SNARE domain mutants revealed a second signal within the SNARE motif (between layers -4 and -1) that was required for efficient ER export. Other SNARE domain mutants that contained this signal were efficiently packaged into COPII vesicles but failed to assemble into SNARE complexes. Together these results indicated that SNARE complex formation is neither required nor sufficient for Sec22p packaging into COPII transport vesicles and subsequent targeting to the Golgi complex. We propose that the COPII budding machinery has a preference for unassembled ER/Golgi SNARE proteins. PMID- 15123694 TI - The Pseudomonas syringae genome encodes a combined mannuronan C-5-epimerase and O acetylhydrolase, which strongly enhances the predicted gel-forming properties of alginates. AB - Alginates are industrially important, linear copolymers of beta-d-mannuronic acid (M) and its C-5-epimer alpha-l-guluronic acid (G). The G residues originate from a postpolymerization reaction catalyzed by mannuronan C-5-epimerases (MEs), leading to extensive variability in M/G ratios and distribution patterns. Alginates containing long continuous stretches of G residues (G blocks) can form strong gels, a polymer type not found in alginate-producing bacteria belonging to the genus Pseudomonas. Here we show that the Pseudomonas syringae genome encodes a Ca(2+)-dependent ME (PsmE) that efficiently forms such G blocks in vitro. The deduced PsmE protein consists of 1610 amino acids and is a modular enzyme related to the previously characterized family of Azotobacter vinelandii ME (AlgE1-7). A- and R-like modules with sequence similarity to those in the AlgE enzymes are found in PsmE, and the A module of PsmE (PsmEA) was found to be sufficient for epimerization. Interestingly, an R module from AlgE4 stimulated Ps-mEA activity. PsmE contains two regions designated M and RTX, both presumably involved in the binding of Ca(2+). Bacterial alginates are partly acetylated, and such modified residues cannot be epimerized. Based on a detailed computer-assisted analysis and experimental studies another PsmE region, designated N, was found to encode an acetylhydrolase. By the combined action of N and A PsmE was capable of redesigning an extensively acetylated alginate low in G from a non gel-forming to a gel-forming state. Such a property has to our knowledge not been previously reported for an enzyme acting on a polysaccharide. PMID- 15123695 TI - Hetero-oligomerization between beta2- and beta3-adrenergic receptors generates a beta-adrenergic signaling unit with distinct functional properties. AB - The ability of the closely related beta(2)- and beta(3)-adrenergic receptors (AR) to form hetero-oligomers was assessed by bioluminescence resonance energy transfer. Quantitative bioluminescence resonance energy transfer titration curves revealed that the beta(2)AR has identical propensity to hetero-oligomerize with the beta(3)AR than to form homo-oligomers. To determine the influence of heterooligomerization, a HEK293 cell line stably expressing an excess of beta(3)AR over beta(2)AR was generated so that all beta(2)AR are engaged in hetero-oligomerization with beta(3)AR, providing a tool to study the effect of hetero-oligomerization on beta(2)AR function in the absence of any beta(2)AR homooligomer. The hetero-oligomerization had no effect on the ligand binding properties of various beta(2)AR ligands and did not affect the potency of isoproterenol to stimulate adenylyl cyclase. Despite the unaltered ligand binding properties of the beta(2/3)AR hetero-oligomer, the stable association of the beta(2)AR with the beta(3)AR completely blocked agonist-stimulated internalization of the beta(2)AR. Given that the beta(3)AR is resistant to agonist-promoted endocytosis, the results indicate that the beta(3)AR acted as a dominant negative of the beta(2)AR endocytosis process. Consistent with this notion, the beta(2/3)AR hetero-oligomer displayed a lower propensity to recruit beta-arrestin-2 than the beta(2)AR. The hetero-oligomerization also led to a change in G protein coupling selectivity. Indeed, in contrast to beta(2)AR and beta(3)AR, which regulate adenylyl cyclase and extracellular signal-regulated kinase activity through a coupling to G(s) and G(i/o), no G(i/o) coupling was observed for the beta(2/3)AR hetero-oligomer. Together, these results demonstrate that hetero-oligomerization between beta(2)AR and beta(3)AR forms a beta adrenergic signaling unit that possesses unique functional properties. PMID- 15123696 TI - S-glutathiolation of Ras mediates redox-sensitive signaling by angiotensin II in vascular smooth muscle cells. AB - Angiotensin II (AII) increases production of reactive oxygen species from NAD(P)H oxidase, a response that contributes to vascular hypertrophy. Here we show in cultured vascular smooth muscle cells that S-glutathiolation of the redox sensitive Cys(118) on the small GTPase, Ras, plays a critical role in AII-induced hypertrophic signaling. AII simultaneously increased the Ras activity and the S glutathiolation of Ras (GSS-Ras) detected by biotin-labeled GSH or mass spectrometry. Both the increase in activity and GSS-Ras was labile under reducing conditions, suggesting the essential nature of this thiol modification to Ras activation. Overexpression of catalase, a dominant-negative p47(phox), or glutaredoxin-1 decreased GSS-Ras, Ras activation, p38, and Akt phosphorylation and the induction of protein synthesis by AII. Furthermore, expression of a Cys(118) mutant Ras decreased AII-mediated p38 and Akt phosphorylation as well as protein synthesis. These results show that H(2)O(2) from NAD(P)H oxidase forms GSS-Ras on Cys(118) and increases its activity leading to p38 and Akt phosphorylation, which contributes to the induction of protein synthesis. This study suggests that GSS-Ras is a redox-sensitive signaling switch that participates in the cellular response to AII. PMID- 15123697 TI - Plasma membrane expression of T-type calcium channel alpha(1) subunits is modulated by high voltage-activated auxiliary subunits. AB - It has been suggested that the auxiliary subunits of high voltage-activated (HVA) calcium channels modulate T-type, low voltage-activated (LVA) calcium channels. Such a regulation has yet to be documented, especially because there has been no biochemical characterization of T-channels. To monitor total protein levels and plasma membrane expression of T-channels in living cells, external epitopes (hemagglutinin, FLAG) were introduced into human recombinant Ca(V)3 channels that were also N-terminally fused to green fluorescent protein. Utilizing Western blot techniques, fluorescence flow cytometry, immunofluorescence, luminometry, and electrophysiology, we describe here that beta(1b) and alpha(2)-delta(1) subunits enhance the level of Ca(V)3 proteins as well as their plasma membrane expression in various expression systems. We also report that, in both Xenopus oocytes and mammalian cells, the alpha(2)-delta(1) subunits increase by at least and beta(1b) 2-fold the current density of Ca(V)3 channels with no change in the electrophysiological properties. Altogether, these data indicate that HVA auxiliary subunits modulate Ca(V)3 channel surface expression, suggesting that the membrane targeting of HVA and LVA alpha(1) subunits is regulated dynamically through the expression of a common set of regulatory subunits. PMID- 15123698 TI - Regulation of the mammalian circadian clock by cryptochrome. PMID- 15123699 TI - The fourth transmembrane segment of the Na,K-ATPase alpha subunit: a systematic mutagenesis study. AB - The Na,K-ATPase is a major ion-motive ATPase of the P-type family responsible for many aspects of cellular homeostasis. To determine the structure of the pathway for cations across the transmembrane portion of the Na,K-ATPase, we mutated 24 residues of the fourth transmembrane segment into cysteine and studied their function and accessibility by exposure to the sulfhydryl reagent 2-aminoethyl methanethiosulfonate. Accessibility was also examined after treatment with palytoxin, which transforms the Na,K-pump into a cation channel. Of the 24 tested cysteine mutants, seven had no or a much reduced transport function. In particular cysteine mutants of the highly conserved "PEG" motif had a strongly reduced activity. However, most of the non-functional mutants could still be transformed by palytoxin as well as all of the functional mutants. Accessibility, determined as a 2-aminoethyl-methanethiosulfonate-induced reduction of the transport activity or as inhibition of the membrane conductance after palytoxin treatment, was observed for the following positions: Phe(323), Ile(322), Gly(326), Ala(330), Pro(333), Glu(334), and Gly(335). In accordance with a structural model of the Na,K-ATPase obtained by homology modeling with the two published structures of sarcoplasmic and endoplasmic reticulum calcium ATPase (Protein Data Bank codes 1EUL and 1IWO), the results suggest the presence of a cation pathway along the side of the fourth transmembrane segment that faces the space between transmembrane segments 5 and 6. The phenylalanine residue in position 323 has a critical position at the outer mouth of the cation pathway. The residues thought to form the cation binding site II ((333)PEGL) are also part of the accessible wall of the cation pathway opened by palytoxin through the Na,K pump. PMID- 15123700 TI - Hindering the strand passage reaction of human topoisomerase IIalpha without disturbing DNA cleavage, ATP hydrolysis, or the operation of the N-terminal clamp. AB - DNA topoisomerase II is an essential enzyme that releases a topological strain in DNA by introduction of transient breaks in one DNA helix through which another helix is passed. While changing DNA topology, ATP is required to drive the enzyme through a series of conformational changes dependent on interdomain communication. We have characterized a human topoisomerase IIalpha enzyme with a two-amino acid insertion at position 351 in the transducer domain. The mutation specifically abolishes the DNA strand passage event of the enzyme, probably because of a sterical hindrance of T-segment transport. Thus, the enzyme fails to decatenate and relax DNA, even though it is fully capable of ATP hydrolysis, closure of the N-terminal clamp, and DNA cleavage. The cleavage activity is increased, suggesting that the transducer domain has a role in regulating DNA cleavage. Furthermore, the enzyme has retained a tendency to increase DNA cleavage upon nucleotide binding and also responds to DNA with elevated ATP hydrolysis. However, the DNA-mediated increase in ATP hydrolysis is lower than that obtained with the wild-type enzyme but similar to that of a cleavage deficient topoisomerase IIalpha enzyme. Our results strongly suggest that the strand passage event is required for efficient DNA stimulation of topoisomerase II-mediated ATP hydrolysis, whereas the stimulation occurs independent of the DNA cleavage reaction per se. A comparison of the strand passage deficient-enzyme described here and the cleavage-deficient enzyme may have applications in other studies where a clear distinction between strand passage and topoisomerase II mediated DNA cleavage is desirable. PMID- 15123701 TI - Transcription of the yeast iron regulon does not respond directly to iron but rather to iron-sulfur cluster biosynthesis. AB - Saccharomyces cerevisiae responds to iron deprivation by increased transcription of the iron regulon, including the high affinity cell-surface transport system encoded by FET3 and FTR1. Here we demonstrate that transcription of these genes does not respond directly to cytosolic iron but rather to the mitochondrial utilization of iron for the synthesis of iron-sulfur (Fe-S) clusters. We took advantage of a mutant form of an iron-dependent enzyme in the sterol pathway (Erg25-2p) to assess cytosolic iron levels. We showed that disruption of mitochondrial Fe-S biosynthesis, which results in excessive mitochondrial iron accumulation, leads to transcription of the iron transport system independent of the cytosolic iron level. There is an inverse correlation between the activity of the mitochondrial Fe-S-containing enzyme aconitase and the induction of FET3. Regulation of transcription by Fe-S biosynthesis represents a mechanism by which cellular iron acquisition is integrated with mitochondrial iron metabolism. PMID- 15123702 TI - Bt-R1a extracellular cadherin repeat 12 mediates Bacillus thuringiensis Cry1Ab binding and cytotoxicity. AB - The cadherin protein Bt-R(1a) is a receptor for Bacillus thuringiensis Cry1A toxins in Manduca sexta. Cry1Ab toxin is reported to bind specific epitopes located in extracellular cadherin repeat (CR) 7 and CR11 on Bt-R(1) (Gomez, B., Miranda-Rios, J., Riudino-Pinera, E., Oltean, D. I., Gill, S. S., Bravo, A., and Soberon, M. (2002) J. Biol. Chem. 277, 30137-30143; Dorsch, J. A., Candas, M., Griko, N., Maaty, W., Midboe, E., Vadlamudi, R., and Bulla, L. (2002) Insect Biochem. Mol. Biol. 32, 1025-1036). We transiently expressed CR domains of Bt R(1a) in Drosophila melanogaster Schneider 2 (S2) cells as fusion peptides between a signal peptide and a terminal region that included membrane-proximal, membrane-spanning, and cytoplasmic domains. A domain consisting of CR11 and 12 was the minimal (125)I-Cry1Ab binding region detected under denaturing conditions. Only CR12 was essential for Cry1Ab binding and cytotoxicity to S2 cells when tested under native conditions. Under these conditions expressed CR12 bound (125)I-Cry1Ab with high affinity (K(com) = 2.9 nm). Flow cytometry assays showed that expression of CR12 conferred susceptibility to Cry1Ab in S2 cells. Derivatives of Bt-R(1a) with separate deletions of CR7, 11, and 12 were expressed in S2 cells. Only deletion of CR12 caused loss of Cry1Ab binding and cytotoxicity. These results demonstrate that CR12 is the essential Cry1Ab binding component on Bt-R(1) that mediates Cry1Ab-induced cytotoxicity. PMID- 15123703 TI - Interaction of SDS with Na+/K+-ATPase: SDS-solubilized enzyme retains partial structure and function. AB - Because nearly all structure/function studies on Na(+)/K(+)-ATPase have been done on enzymes prepared in the presence of SDS, we have studied previously unrecognized consequences of SDS interaction with the enzyme. When the purified membrane-bound kidney enzyme was solubilized with SDS or TDS concentrations just sufficient to cause complete solubilization, but not at concentrations severalfold higher, the enzyme retained quaternary structure, exhibiting alpha,alpha-, alpha,beta-, beta,beta-, and alpha,gamma-associations as detected by chemical cross-linking. The presence of solubilized oligomers was confirmed by sucrose density gradient centrifugation. This solubilized enzyme had no ATPase activity and was not phosphorylated by ATP, but it retained the ability to occlude Rb(+) and Na(+). This, and comparison of cross-linking patterns obtained with different reagents, suggested that the transmembrane domains of the enzyme are more resistant to SDS-induced unfolding than its other domains. These findings (a). indicate that the partially unfolded oligomer(s) retaining partial function is the intermediate in the SDS-induced denaturation of the native membrane enzyme having the minimum oligomeric structure of (alpha,beta,gamma)(2) and (b). suggest potential functions for Na(+)/K(+)-ATPase with intrinsically unfolded domains. Mixtures of solubilized/partially unfolded enzyme and membrane bound enzyme exhibited cross-linking patterns and Na(+) occlusion capacities different from those of either enzyme species, suggesting that the two interact. Formation of the partially unfolded enzyme during standard purification procedure for the preparation of the membrane-bound enzyme was shown, indicating that it is necessary to ensure the separation of the partially unfolded enzyme from the membrane-bound enzyme to avoid the distortion of the properties of the latter. PMID- 15123704 TI - Phosphatidylinositol 3-kinase signaling is involved in neurogenesis during Xenopus embryonic development. AB - Phosphatidylinositol 3-kinase (PI3K) has numerous cellular functions, including cell survival and proliferation. In this study, we demonstrated that the expression of the active form of PI3K induced dorsal differentiation and axis duplication and strongly induced the expression of neural markers. In contrast, the inhibition of PI3K activity by its dominant negative mutant induced the phenotype of losing posterior structures and the expression of ventral markers. Akt is an essential target of PI3K for neurogenesis. The expression of the active form of Akt induced axis duplication and increased the expression of neural markers. Inhibition of the Akt activity abolished the PI3K-induced double heads and axes. This signal transmits through its target, glycogen synthase kinase 3beta, which is known to mediate Wnt signaling for Xenopus development. These results identify a new function of PI3K/Akt signaling in axis formation and neurogenesis during Xenopus embryonic development and provide a direct link between growth factor-mediated PI3K/Akt signaling and Wnt signaling during embryonic development. PMID- 15123705 TI - Reactive oxygen species mediate Met receptor transactivation by G protein-coupled receptors and the epidermal growth factor receptor in human carcinoma cells. AB - Cross-communication between the Met receptor tyrosine kinase and the epidermal growth factor receptor (EGFR) has been proposed to involve direct association of both receptors and EGFR kinase-dependent phosphorylation. Here, we demonstrate that in human hepatocellular and pancreatic carcinoma cells the Met receptor becomes tyrosine phosphorylated not only upon EGF stimulation but also in response to G protein-coupled receptor (GPCR) agonists. Whereas specific inhibition of the EGFR kinase activity blocked EGF- but not GPCR agonist-induced Met receptor transactivation, it was abrogated in the presence of a reducing agent or treatment of cells with a NADPH oxidase inhibitor. Both GPCR ligands and EGF are further shown to increase the level of reactive oxygen species within the cell. Interestingly, stimulation of the Met receptor by either GPCR agonists, EGF or its cognate ligand HGF, resulted in release of Met-associated beta-catenin and in its Met-dependent translocation into the nucleus, as analyzed by small interfering RNA-mediated knockdown of the Met receptor. Our results provide a new molecular explanation for cell surface receptor cross-talk involving the Met receptor and thereby link the wide diversity of GPCRs and the EGFR to the oncogenic potential of Met signaling in human carcinoma cells. PMID- 15123706 TI - Trans-inactivation of receptor tyrosine kinases by novel angiotensin II AT2 receptor-interacting protein, ATIP. AB - Negative regulation of mitogenic pathways is a fundamental process that remains poorly characterized. The angiotensin II AT2 receptor is a rare example of a 7 transmembrane domain receptor that negatively cross-talks with receptor tyrosine kinases to inhibit cell growth. In the present study, we report the molecular cloning of a novel protein, ATIP1 (AT2-interacting protein), which interacts with the C-terminal tail of the AT2 receptor, but not with those of other receptors such as angiotensin AT1, bradykinin BK2, and adrenergic beta(2) receptor. ATIP1 defines a family of at least four members that possess the same domain of interaction with the AT2 receptor, contain a large coiled-coil region, and are able to dimerize. Ectopic expression of ATIP1 in eukaryotic cells leads to inhibition of insulin, basic fibroblast growth factor, and epidermal growth factor-induced ERK2 activation and DNA synthesis, and attenuates insulin receptor autophosphorylation, in the same way as the AT2 receptor. The inhibitory effect of ATIP1 requires expression, but not ligand activation, of the AT2 receptor and is further increased in the presence of Ang II, indicating that ATIP1 cooperates with AT2 to transinactivate receptor tyrosine kinases. Our findings therefore identify ATIP1 as a novel early component of growth inhibitory signaling cascade. PMID- 15123707 TI - Cytoplasmic nuclear transfer of the actin-capping protein tropomodulin. AB - Tropomodulin (Tmod) is a cytoskeletal actin-capping protein that interacts with tropomyosin at the pointed end of actin filaments. E-Tmod is an isoform that expresses predominantly in cardiac cells and slow skeletal muscle fibers. We unexpectedly discovered significant levels of Tmod in nuclei and then defined peptide domains in Tmod responsible for nuclear import and export. These domains resemble, and function as, a nuclear export signal (NES) and a pattern 4 nuclear localization signal (NLS). Both motifs are conserved in other Tmod isoforms and across species. Comparisons of wild-type Tmod and Tmod carrying mutations in these peptide domains revealed that Tmod normally traffics through the nucleus. These observations logically presuppose that Tmod functions may include a nuclear role. Indeed, increasing Tmod in the nucleus severely hampered myogenic differentiation and selectively suppressed muscle-specific gene expression (endogenous p21, myosin heavy chain, myogenin, and Tmod) but did not affect endogenous glyceraldehyde-3-phosphate dehydrogenase or expression from a transfected E-GFP vector. These results suggest that, at least in myogenic cells, nuclear Tmod may be involved in the differentiation process. PMID- 15123708 TI - Human deafness mutation of myosin VI (C442Y) accelerates the ADP dissociation rate. AB - The missense mutation of Cys(442) to Tyr of myosin VI causes progressive postlingual sensorineural deafness. Here we report the affects of the C442Y mutation on the kinetics of the actomyosin ATP hydrolysis mechanism and motor function of myosin VI. The largest changes in the kinetic mechanism of ATP hydrolysis produced by the C442Y mutation are about 10-fold increases in the rate of ADP dissociation from both myosin VI and actomyosin VI. The rates of ADP dissociation from acto-C442Y myosin VI-ADP and C442Y myosin VI-ADP are 20-40 times more rapid than the steady state rates and cannot be the rate-limiting steps of the hydrolysis mechanism in the presence or absence of actin. The 2-fold increase in the actin gliding velocity of C442Y compared with wild type (WT) may be explained at least in part by the more rapid rate of ADP dissociation. The C442Y myosin VI has a significant increase ( approximately 10-fold) in the steady state ATPase rate in the absence of actin relative to WT myosin VI. The steady state rate of actin-activated ATP hydrolysis is unchanged by the C442Y mutation at low (<10(-7) m) calcium but is calcium-sensitive with a 1.6-fold increase at high ( approximately 10(-4) m) calcium that does not occur with WT. The actin gliding velocity of the C442Y mutant decreases significantly at low surface density of myosin VI, suggesting that the mutation hampers the processive movement of myosin VI. PMID- 15123709 TI - A femtomolar acting octapeptide interacts with tubulin and protects astrocytes against zinc intoxication. AB - An octapeptide was previously described that protects neurons against a wide variety of insults directly and indirectly as a result of interactions (at femtomolar concentrations) with supporting glial cells. The current study set out to identify the octapeptide binding molecules so as to understand the high affinity mechanisms of cellular protection. Studies utilizing affinity chromatography of brain extracts identified tubulin, the brain major protein, as the octapeptide-binding ligand. Dot blot analysis with pure tubulin and the biotinylated octapeptide verified this finding. When added to cerebral cortical astrocytes, the octapeptide (10(-15)-10(-10) m) induced a rapid microtubule reorganization into distinct microtubular structures that were stained by monoclonal tubulin antibodies and visualized by confocal microscopy. Fluorescein labeled octapeptide induced a similar change and was detected in the intracellular milieu, even when cells were incubated at 4 degrees C or at low pH. In a cell-free system, the octapeptide stimulated tubulin assembly into microtubules. Furthermore, treatment of astrocytes with zinc chloride resulted in microtubule disassembly and cell death that was protected by the octapeptide. In conclusion, the results suggest that the octapeptide crosses the plasma membrane and interacts directly with tubulin, the microtubule subunit, to induce microtubule reorganization and improved survival. Because microtubules are the key component of the neuronal and glial cytoskeleton that regulates cell division, differentiation, and protection, this finding may explain the breadth and efficiency of the cellular protective capacities of the octapeptide. PMID- 15123710 TI - Structural basis for the autoinhibition and STI-571 inhibition of c-Kit tyrosine kinase. AB - The activity of the c-Kit receptor protein-tyrosine kinase is tightly regulated in normal cells, whereas deregulated c-Kit kinase activity is implicated in the pathogenesis of human cancers. The c-Kit juxtamembrane region is known to have an autoinhibitory function; however the precise mechanism by which c-Kit is maintained in an autoinhibited state is not known. We report the 1.9-A resolution crystal structure of native c-Kit kinase in an autoinhibited conformation and compare it with active c-Kit kinase. Autoinhibited c-Kit is stabilized by the juxtamembrane domain, which inserts into the kinase-active site and disrupts formation of the activated structure. A 1.6-A crystal structure of c-Kit in complex with STI-571 (Imatinib or Gleevec) demonstrates that inhibitor binding disrupts this natural mechanism for maintaining c-Kit in an autoinhibited state. Together, these results provide a structural basis for understanding c-Kit kinase autoinhibition and will facilitate the structure-guided design of specific inhibitors that target the activated and autoinhibited conformations of c-Kit kinase. PMID- 15123711 TI - Regulation of the mouse epithelial Ca2(+) channel TRPV6 by the Ca(2+)-sensor calmodulin. AB - TRPV5 and TRPV6 are members of the superfamily of transient receptor potential (TRP) channels and facilitate Ca(2+) influx in a variety of epithelial cells. The activity of these Ca(2+) channels is tightly controlled by the intracellular Ca(2+) concentration in close vicinity to the channel mouth. The molecular mechanism underlying the Ca(2+)-dependent activity of TRPV5/TRPV6 is, however, still unknown. Here, the putative role of calmodulin (CaM) as the Ca(2+) sensor mediating the regulation of channel activity was investigated. Overexpression of Ca(2+)-insensitive CaM mutants (CaM(1234) and CaM(34)) significantly reduced the Ca(2+) as well as the Na(+) current of TRPV6- but not that of TRPV5-expressing HEK293 cells. By combining pull-down assays and co-immunoprecipitations, we demonstrated that CaM binds to both TRPV5 and TRPV6 in a Ca(2+)-dependent fashion. The binding of CaM to TRPV6 was localized to the transmembrane domain (TRPV6(327-577)) and consensus CaM-binding motifs located in the N (1-5-10 motif, TRPV6(88-97)) and C termini (1-8-14 motif, TRPV6(643-656)), suggesting a mechanism of regulation involving multiple interaction sites. Subsequently, chimeric TRPV6/TRPV5 proteins, in which the N and/or C termini of TRPV6 were substituted by that of TRPV5, were co-expressed with CaM(34) in HEK293 cells. Exchanging, the N and/or the C termini of TRPV6 by that of TRPV5 did not affect the CaM(34)-induced reduction of the Ca(2+) and Na(+) currents. These results suggest that CaM positively affects TRPV6 activity upon Ca(2+) binding to EF hands 3 and 4, located in the high Ca(2+) affinity CaM C terminus, which involves the N and C termini and the transmembrane domain of TRPV6. PMID- 15123712 TI - The solution structure of the N-terminal domain of human vitronectin: proximal sites that regulate fibrinolysis and cell migration. AB - The three-dimensional structure of an N-terminal fragment comprising the first 51 amino acids from human plasma vitronectin, the somatomedin B (SMB) domain, has been determined by two-dimensional NMR approaches. An average structure was calculated, representing the overall fold from a set of 20 minimized structures. The core residues (18-41) overlay with a root mean square deviation of 2.29 +/- 0.62 A. The N- and C-terminal segments exhibit higher root mean square deviations, reflecting more flexibility in solution and/or fewer long-range NOEs for these regions. Residues 26-30 form a unique single-turn alpha-helix, the locus where plasminogen activator inhibitor type-1 (PAI-1) is bound. This structure of this helix is highly homologous with that of a recombinant SMB domain solved in a co-crystal with PAI-1 (Zhou, A., Huntington, J. A., Pannu, N. S., Carrell, R. W., and Read, R. J. (2003) Nat. Struct. Biol. 10, 541-544), although the remainder of the structure differs. Significantly, the pattern of disulfide cross-links observed in this material isolated from human plasma is altogether different from the disulfides proposed for recombinant forms. The NMR structure reveals the relative orientation of binding sites for cell surface receptors, including an integrin-binding site at residues 45-47, which was disordered and did not diffract in the co-crystal, and a site for the urokinase receptor, which overlaps with the PAI-1-binding site. PMID- 15123713 TI - Solution structure of spheniscin, a beta-defensin from the penguin stomach. AB - Recently two beta-defensins, named spheniscins, have been isolated from the stomach content of the king penguin (Aptenodytes patagonicus), which is capable of preserving food for several weeks during egg incubation (Thouzeau, C., Le Maho, Y., Froget, G., Sabatier, L., Le Bohec, C., Hoffmann, J. A., and Bulet, P. (2003) J. Biol. Chem. 278, 51053-51058). It has been proposed that, in combination with other antimicrobial peptides, spheniscins may be involved in this long term preservation of food in the bird's stomach. To draw some structure/function features, the three-dimensional structure in aqueous solution of the most abundant spheniscin (Sphe-2) was determined by two-dimensional NMR and molecular modeling techniques. The overall fold of Sphe-2 includes a three stranded antiparallel beta-sheet stabilized by three disulfide bridges with a pairing typical of beta-defensins. In addition, the N-terminal segment shows helical features on most structures. Sphe-2 is highly cationic, and its surface displays a hydrophobic patch. Comparative modeling revealed that this patch is preserved in avian defensins. The activity of Sphe-2 against a pathogenic Gram positive strain was retained in vitro in the conditions of osmolarity found in penguin stomach content and also in different salt concentrations and compositions up to those reported for seawater. Comparison with structurally related mammalian beta-defensins showed that the hydrophobic patch is not preserved in mammalian beta-defensins and that the high cationicity of Sphe-2 is presumably the critical factor for its retained activity in high salt concentrations. Such peculiarities, in addition to a broad activity spectrum, suggest that penguin defensins may represent interesting probes for the design of highly efficient antibiotics to fight off pathogens that develop in relatively salt-rich body fluids. PMID- 15123714 TI - PKD2 interacts and co-localizes with mDia1 to mitotic spindles of dividing cells: role of mDia1 IN PKD2 localization to mitotic spindles. AB - Mutations in pkd2 result in the type 2 form of autosomal dominant polycystic kidney disease, which accounts for approximately 15% of all cases of the disease. PKD2, the protein product of pkd2, belongs to the transient receptor potential superfamily of cation channels, and it can function as a mechanosensitive channel in the primary cilium of kidney cells, an intracellular Ca(2+) release channel in the endoplasmic reticulum, and/or a nonselective cation channel in the plasma membrane. We have identified mDia1/Drf1 (mammalian Diaphanous or Diaphanous related formin 1 protein) as a PKD2-interacting protein by yeast two-hybrid screen. mDia1 is a member of the RhoA GTPase-binding formin homology protein family that participates in cytoskeletal organization, cytokinesis, and signal transduction. We show that mDia1 and PKD2 interact in native and in transfected cells, and binding is mediated by the cytoplasmic C terminus of PKD2 binding to the mDia1 N terminus. The interaction is more prevalent in dividing cells in which endogenous PKD2 and mDia1 co-localize to the mitotic spindles. RNA interference experiments reveal that endogenous mDia1 knockdown in HeLa cells results in the loss of PKD2 from mitotic spindles and alters intracellular Ca(2+) release. Our results suggest that mDia1 facilitates the movement of PKD2 to a centralized position during cell division and has a positive effect on intracellular Ca(2+) release during mitosis. This may be important to ensure equal segregation of PKD2 to the daughter cell to maintain a necessary level of channel activity. Alternatively, PKD2 channel activity may be important in the cell division process or in cell fate decisions after division. PMID- 15123715 TI - Control of replication origin density and firing time in Xenopus egg extracts: role of a caffeine-sensitive, ATR-dependent checkpoint. AB - A strict control of replication origin density and firing time is essential to chromosomal stability. Replication origins in early frog embryos are located at apparently random sequences, are spaced at close ( approximately 10-kb) intervals, and are activated in clusters that fire at different times throughout a very brief S phase. Using molecular combing of DNA from sperm nuclei replicating in Xenopus egg extracts, we show that the temporal order of origin firing can be modulated by the nucleocytoplasmic ratio and the checkpoint abrogating agent caffeine in the absence of external challenge. Increasing the concentration of nuclei in the extract increases S phase length. Contrary to a previous interpretation, this does not result from a change in local origin spacing but from a spreading of the time over which distinct origin clusters fire and from a decrease in replication fork velocity. Caffeine addition or ATR inhibition with a specific neutralizing antibody increases origin firing early in S phase, suggesting that a checkpoint controls the time of origin firing during unperturbed S phase. Furthermore, fork progression is impaired when excess forks are assembled after caffeine treatment. We also show that caffeine allows more early origin firing with low levels of aphidicolin treatment but not higher levels. We propose that a caffeine-sensitive, ATR-dependent checkpoint adjusts the frequency of initiation to the supply of replication factors and optimizes fork density for safe and efficient chromosomal replication during normal S phase. PMID- 15123716 TI - Dissecting the cell-killing mechanism of the topoisomerase II-targeting drug ICRF 193. AB - Topoisomerase II is an essential enzyme that is targeted by a number of clinically valuable anticancer drugs. One class referred to as topoisomerase II poisons works by increasing the cellular level of topoisomerase II-mediated DNA breaks, resulting in apoptosis. Another class of topoisomerase II-directed drugs, the bis-dioxopiperazines, stabilizes the conformation of the enzyme where it attains an inactive salt-stable closed clamp structure. Bis-dioxopiperazines, similar to topoisomerase II poisons, induce cell killing, but the underlying mechanism is presently unclear. In this study, we use three different biochemically well characterized human topoisomerase IIalpha mutant enzymes to dissect the catalytic requirements needed for the enzyme to cause dominant sensitivity in yeast to the bis-dioxopirazine ICRF-193 and the topoisomerase II poison m-AMSA. We find that the clamp-closing activity, the DNA cleavage activity, and even both activities together are insufficient for topoisomerase II to cause dominant sensitivity to ICRF-193 in yeast. Rather, the strand passage event per se is an absolute requirement, most probably because this involves a simultaneous interaction of the enzyme with two DNA segments. Furthermore, we show that the ability of human topoisomerase IIalpha to cause dominant sensitivity to m-AMSA in yeast does not depend on clamp closure or strand passage but is directly related to the capability of the enzyme to respond to m-AMSA with increased DNA cleavage complex formation. PMID- 15123717 TI - Tissue inhibitor of metalloproteinases-3 (TIMP-3) is a binding partner of epithelial growth factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1). Implications for macular degenerations. AB - Tissue inhibitor of metalloproteinases-3 (TIMP-3) is a matrix-bound inhibitor of matrix metalloproteinases. Mutations in the Timp-3 gene cause Sorsby fundus dystrophy (SFD), a hereditary macular degenerative disease. The pathogenic mechanisms responsible for the disease phenotype are unknown. In an in vivo quest for binding partners of the TIMP-3 protein in the subretina, we identified epidermal growth factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1, also known as fibulin 3) as a strong interacting protein. The COOH terminal end of TIMP-3 was involved in the interaction. Interestingly, a missense mutation in EFEMP1 is responsible for another hereditary macular degenerative disease, Malattia Leventinese (ML). Both SFD and ML have strong similarities to age-related macular degeneration (AMD), a major cause of blindness in the elderly population of the Western hemisphere. Our results were supported by significant accumulation and expression overlap of both TIMP-3 and EFEMP1 between the retinal pigment epithelia and Bruch membrane in the eyes of ML and AMD patients. These results provide the first link between two different macular degenerative disease genes and imply the possibility of a common pathogenic mechanism behind different forms of macular degeneration. PMID- 15123718 TI - Conformation of membrane-associated proapoptotic tBid. AB - The proapoptotic Bcl-2 family protein Bid is cleaved by caspase-8 to release the C-terminal fragment tBid, which translocates to the outer mitochondrial membrane and induces massive cytochrome c release and cell death. In this study, we have characterized the conformation of tBid in lipid membrane environments, using NMR and CD spectroscopy with lipid micelle and lipid bilayer samples. In micelles, tBid adopts a unique helical conformation, and the solution NMR (1)H/(15)N HSQC spectra have a single well resolved resonance for each of the protein amide sites. In lipid bilayers, tBid associates with the membrane with its helices parallel to the membrane surface and without trans-membrane helix insertion, and the solid-state NMR (1)H/(15)N polarization inversion with spin exchange at the magic angle spectrum has all of the amide resonances centered at (15)N chemical shift (70-90 ppm) and (1)H-(15)N dipolar coupling (0-5 kHz) frequencies associated with NH bonds parallel to the bilayer surface, with no intensity at frequencies associated with NH bonds in trans-membrane helices. Thus, the cytotoxic activity of tBid at mitochondria may be similar to that observed for antibiotic polypeptides, which bind to the surface of bacterial membranes as amphipathic helices and destabilize the bilayer structure, promoting the leakage of cell contents. PMID- 15123719 TI - Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit modulate RAG mediated cleavage: implications for the enforcement of the 12/23 rule. AB - The 12/23 rule is a critical step for regulation of V(D)J recombination. To date, only the RAG proteins and high mobility group protein 1 or 2 have been implicated in 12/23 regulation. Through protein fractionation and biochemical experiments, we find that Ku70/Ku80 and DNA-dependent protein kinase catalytic subunit (DNA PKcs) modulate RAG-mediated cleavage. Modulation of cleavage by Ku70/80 and DNA PKcs results in preferential inhibition of 12/12 and 23/23 DNA cleavage, thus increasing 12/23 rule specificity. This observation indicates that DNA repair factors, Ku70/80 and DNA-PKcs, might be present upstream of the DNA cleavage events and not recruited downstream as is currently thought, assigning new nonrepair functions to the DNA-dependent protein kinase. PMID- 15123720 TI - Post-transcriptional regulation of sterol regulatory element-binding protein-1 by ethanol induces class I alcohol dehydrogenase in rat liver. AB - Members of the sterol regulatory element-binding protein (SREBP) family of transcription factors control the synthesis and uptake of cholesterol, fatty acids, triglycerides, and phospholipids. Continuous intragastric infusion of ethanol-containing diets as part of total enteral nutrition generates well defined 6-day cycles (pulses) of urine ethanol concentrations (UECs) in rats. Pulsatile UECs are the result of cyclical expression and activity of the principal alcohol-metabolizing enzyme, hepatic Class I alcohol dehydrogenase (ADH), and this mechanism involves regulated CCAAT/enhancer-binding protein-beta expression and binding to the ADH promoter. In this study, we further explore the molecular mechanism for ethanol-induced ADH expression during the UEC pulse in adult male rats fed ethanol by total enteral nutrition for 21-30 days. In hypophysectomized rats, in which the ADH protein increased by approximately 6 fold, the nuclear form of SREBP-1 decreased by approximately 7-fold. Because the ADH promoter contains two canonical sterol response element (SRE) sites (-63 to 53 and -52 to -40 relative to the transcription start site), electrophoretic mobility shift assays were conducted using an ADH-specific SRE site. Hepatic nuclear protein binding decreased by 2.4-fold on the ascending limbs and by 3.6 fold on the descending limbs of UEC pulses (p < 0.05). The specificity of nuclear protein binding to the ADH-SRE site was confirmed using antibody and UV cross link assays. The in vivo binding status of SREBP-1 to ADH-SRE sites, as measured by the chromatin immunoprecipitation assay, had a pattern very similar to the electrophoretic mobility shift assay results. Functional analysis of the ADH-SREs demonstrated these sites to be essential for ADH transcription. In vitro transcription assays demonstrated that depletion of the SREBP-1 protein from nuclear extracts increased transcription activity by approximately 5-fold and that the liver X receptor agonist T0901317 (a known activator of SREBP-1c transcription) reduced in vitro expression of ADH mRNA by 2-fold. We conclude that SREBP-1 is a negative regulator of the ADH gene and may work in concert with the CCAAT/enhancer-binding proteins to mediate ethanol induction of ADH in vivo. PMID- 15123721 TI - Hydrogen peroxide activates the Gas6-Axl pathway in vascular smooth muscle cells. AB - Axl, a receptor tyrosine kinase, is involved in cell survival, proliferation, and migration. We have shown that Axl expression increases in the neointima of balloon-injured rat carotids. Because oxidative stress is known to play a major role in remodeling of injured vessels, we hypothesized that H(2)O(2) might activate Axl by promoting autophosphorylation. H(2)O(2) rapidly stimulated Axl tyrosine phosphorylation in rat vascular smooth muscle cells within 1 min that was maximal at 5 min (6-fold). The response to H(2)O(2) was concentration dependent with EC(50) of approximately 500 microm. Axl phosphorylation was partly dependent on production of its endogenous ligand, growth arrest gene 6 (Gas6), because Axl-Fc, a fragment of Axl extracellular domain that neutralizes Gas6, inhibited H(2)O(2)-induced Axl phosphorylation by 50%. Axl phosphorylation by H(2)O(2) was also attenuated by warfarin, which inhibits Gas6 activity by preventing post-translational modification. In intact vessels Axl was phosphorylated by H(2)O(2), and Axl phosphorylation was inhibited by warfarin treatment in balloon-injured carotids. Akt, a downstream target of Axl, was phosphorylated by H(2)O(2)in Axl(+/+) mouse aorta but significantly inhibited in Axl(-/-) aorta. Intimal proliferation was decreased significantly in a cuff injury model in Axl(-/-) mice compared with Axl(+/+) mice. In summary, Axl is an important signaling mediator for oxidative stress in cultured vascular smooth muscle cells and intact vessels and may represent an important therapeutic target for vascular remodeling and response to injury. PMID- 15123722 TI - Ectodomain shedding of SHPS-1 and its role in regulation of cell migration. AB - SHPS-1 is a transmembrane protein whose cytoplasmic region undergoes tyrosine phosphorylation and then binds the protein-tyrosine phosphatase SHP-2. Formation of the SHPS-1-SHP-2 complex is implicated in regulation of cell migration. In addition, SHPS-1 and its ligand CD47 constitute an intercellular recognition system that contributes to inhibition of cell migration by cell-cell contact. The ectodomain of SHPS-1 has now been shown to be shed from cells in a reaction likely mediated by a metalloproteinase. This process was promoted by activation of protein kinase C or of Ras, and the released ectodomain exhibited minimal CD47 binding activity. Metalloproteinases catalyzed the cleavage of a recombinant SHPS 1-Fc fusion protein in vitro, and the primary cleavage site was localized to the juxtamembrane region of SHPS-1. Forced expression of an SHPS-1 mutant resistant to ectodomain shedding impaired cell migration, cell spreading, and reorganization of the actin cytoskeleton. It also increased the tyrosine phosphorylation of paxillin and FAK triggered by cell adhesion. These results suggest that shedding of the ectodomain of SHPS-1 plays an important role in regulation of cell migration and spreading by this protein. PMID- 15123723 TI - Human constitutive androstane receptor mediates induction of CYP2B6 gene expression by phenytoin. AB - Compared with its rodent orthologs, little is known about the chemical specificity of human constitutive androstane receptor (hCAR) and its regulation of hepatic enzyme expression. Phenytoin (PHY), a widely used antiepileptic drug, is a potent inducer of CYP2B6 in primary human hepatocytes, but does not activate human pregnane X receptor (PXR) significantly in cell-based transfection assays at the same concentrations associated with potent induction of CYP2B6. Based on this observation, we hypothesized that PHY may be a selective activator of hCAR. In primary human hepatocytes, expression of CYP2B6 reporter genes containing phenobarbital-responsive enhancer module (PBREM) or PBREM/xenobiotic-responsive enhancer module (XREM) response elements were activated up to 14- and 28-fold, respectively, by 50 microm PHY. By contrast, parallel experiments in HepG2 cell lines co-transfected with an hPXR expression vector did not show increased reporter activity. These results indicated that a PXR-independent pathway, which is retained in primary hepatocytes, is responsible for PHY induction of CYP2B6. Further experiments revealed that PHY effectively translocates hCAR from the cytoplasm into the nucleus in both primary human hepatocytes and CAR(-/-) mice. Compared with vehicle controls, PHY administration significantly increased CYP2B6 reporter gene expression, when this reporter construct was delivered together with hCAR expression vector into CAR(-/-) mice. However, PHY did not increase reporter gene expression in CAR(-/-) mice in the absence of hCAR vector, implying that CAR is essential for mediating PHY induction of CYP2B6 gene expression. Taken together, these observations demonstrate that, in contrast to most of the known CYP2B6 inducers, PHY is a selective activator of CAR in humans. PMID- 15123724 TI - Phosphatidylinositol 4,5-bisphosphate regulates adipocyte actin dynamics and GLUT4 vesicle recycling. AB - To investigate the potential role of phosphatidylinositol 4, 5-bisphosphate (PI(4,5)P2) in the regulation of actin polymerization and GLUT4 translocation, the type I phosphatidylinositol 4-phosphate 5-kinases (PIP5Ks) were expressed in 3T3L1 adipocytes. In preadipocytes (fibroblasts) PIP5K expression promoted actin polymerization on membrane-bound vesicles to form motile actin comets. In contrast, expression of PIP5K in differentiated 3T3L1 adipocytes resulted in the formation of enlarged vacuole-like structures coated with F-actin, cortactin, dynamin, and N-WASP. Treatment with either latrunculin B (an inhibitor for actin polymerization) or Clostridium difficile toxin B (a general Rho family inhibitor) resulted in a relatively slower disappearance of coated F-actin from these vacuoles, but the vacuoles themselves remained unaffected. Functionally, the increased PI(4,5)P2 levels resulted in an inhibition of transferrin receptor and GLUT4 endocytosis and a slow accumulation of these proteins in the PI(4,5)P2 enriched vacuoles along with the non-clathrin-derived endosome marker (caveolin) and the AP-2 adaptor complex. However, these structures were devoid of early endosome markers (EEA1, clathrin) and the biosynthetic membrane secretory machinery markers p115 (Golgi) and syntaxin 6 (trans-Golgi Network). Taken together, these data demonstrate that PI(4,5)P2 has distinct morphologic and functional properties depending upon specific cell context. In adipocytes, altered PI(4,5)P2 metabolism has marked effects on GLUT4 endocytosis and intracellular vesicle trafficking due to the derangement of actin dynamics. PMID- 15123725 TI - Hepatic nuclear factor 3 and nuclear factor 1 regulate 5-aminolevulinate synthase gene expression and are involved in insulin repression. AB - Although the negative regulation of gene expression by insulin has been widely studied, the transcription factors responsible for the insulin effect are still unknown. The purpose of this work was to explore the molecular mechanisms involved in the insulin repression of the 5-aminolevulinate synthase (ALAS) gene. Deletion analysis of the 5'-regulatory region allowed us to identify an insulin responsive region located at -459 to -354 bp. This fragment contains a highly homologous insulin-responsive (IRE) sequence. By transient transfection assays, we determined that hepatic nuclear factor 3 (HNF3) and nuclear factor 1 (NF1) are necessary for an appropriate expression of the ALAS gene. Insulin overrides the HNF3beta or HNF3beta plus NF1-mediated stimulation of ALAS transcriptional activity. Electrophoretic mobility shift assay and Southwestern blotting indicate that HNF3 binds to the ALAS promoter. Mutational analysis of this region revealed that IRE disruption abrogates insulin action, whereas mutation of the HNF3 element maintains hormone responsiveness. This dissociation between HNF3 binding and insulin action suggests that HNF3beta is not the sole physiologic mediator of insulin-induced transcriptional repression. Furthermore, Southwestern blotting assay shows that at least two polypeptides other than HNF3beta can bind to ALAS promoter and that this binding is dependent on the integrity of the IRE. We propose a model in which insulin exerts its negative effect through the disturbance of HNF3beta binding or transactivation potential, probably due to specific phosphorylation of this transcription factor by Akt. In this regard, results obtained from transfection experiments using kinase inhibitors support this hypothesis. Due to this event, NF1 would lose accessibility to the promoter. The posttranslational modification of HNF3 would allow the binding of a protein complex that recognizes the core IRE. These results provide a potential mechanism for the insulin-mediated repression of IRE-containing promoters. PMID- 15123726 TI - Adiponectin stimulates angiogenesis in response to tissue ischemia through stimulation of amp-activated protein kinase signaling. AB - Obesity is a risk factor for the development of cardiovascular diseases that are associated with impaired angiogenesis. Adiponectin is an adipocyte-specific adipocytokine with anti-atherogenic and anti-diabetic properties, and its plasma levels are reduced in association with obesity-linked diseases. Here, we investigated whether adiponectin regulates angiogenesis in response to tissue ischemia using adiponectin knock-out (KO) mice. Angiogenic repair of ischemic hind limbs was impaired in adiponectin-KO mice compared with wild-type (WT) mice as evaluated by laser Doppler flow method and capillary density analyses. Adenovirus-mediated supplement of adiponectin accelerated angiogenic repair in both adiponectin-KO and WT mice. Intramuscular injection of an adenovirus encoding dominant-negative AMP-activated kinase diminished the improvement in limb perfusion seen in WT mice and abolished the adiponectin-induced enhancement of perfusion. These data indicate that adiponectin can function to stimulate angiogenesis in response to ischemic stress by promoting AMP-activated kinase signaling. Therefore, adiponectin may be useful in the treatment for obesity related vascular deficiency diseases. PMID- 15123727 TI - Combinatorial complexity of 5' alternative acetylcholinesterase transcripts and protein products. AB - To explore the scope and significance of alternate promoter usage and its putative inter-relationship to alternative splicing, we searched expression sequence tags for the 5' region of acetylcholinesterase (ACHE) genes. Three and five novel first exons were identified in human and mouse ACHE genes, respectively. Reverse transcription-PCR and in situ hybridization validated most of the predicted transcripts, and sequence analyses of the corresponding genomic DNA regions suggest evolutionarily conserved promoters for each of the novel exons identified. Distinct tissue specificity and stress-related expression patterns of these exons predict combinatorial complexity with known 3' alternative AChE mRNA transcripts. Unexpectedly one of the 5' exons encodes an extended N terminus in-frame with the known AChE sequence, extending the increased complexity to the protein level. The resultant membrane variant(s), designated N-AChE, is developmentally regulated in human brain neurons and blood mononuclear cells. Alternative promoter usage combined with alternative splicing may thus lead to stress-dependent combinatorial complexity of AChE mRNA transcripts and their protein products. PMID- 15123728 TI - Energetics of the sequence-specific binding of single-stranded DNA by the F factor relaxase domain. AB - Transfer of conjugative plasmids between bacteria requires the activity of relaxases or mobilization proteins. These proteins nick the plasmid in a site- and strand-specific manner prior to transfer of the cut strand from donor to recipient. TraI36, the relaxase domain of TraI from plasmid F factor, binds a single-stranded DNA (ssDNA) oligonucleotide containing an F factor sequence with high affinity and sequence specificity. To better understand the energetics of this interaction, we examined the temperature, salt, and pH dependence of TraI36 recognition. Binding is entropically driven below 25 degrees C and enthalpically driven at higher temperatures. van't Hoff analysis yields an estimated deltaC(P)(0) of binding (-3300 cal x mol(-1) x K(-1)) that is larger and more negative than that observed for most double-stranded DNA (dsDNA)-binding proteins. Based on analyses of circular dichroism data and the crystal structure of the unliganded protein, we attribute the deltaC(P)(0) to both burial of hydrophobic surface area and coupled folding and binding of the protein. The salt dependence of the binding indicates that several ssDNA phosphates are buried in the complex, and the pH dependence of the binding suggests that some of these ssDNA phosphates form ionic interactions with basic residues of the protein. Although data are available for relatively few sequence-specific ssDNA-binding proteins, sufficient differences exist between TraI36 and other proteins to indicate that, like dsDNA-binding proteins, ssDNA-binding proteins use different motifs and combinations of forces to achieve specific recognition. PMID- 15123729 TI - Molecular mechanism of dimerization of Bowman-Birk inhibitors. Pivotal role of ASP76 in the dimerzation. AB - Horsegram (Dolichos biflorus), a protein-rich leguminous pulse, is a crop native to Southeast Asia and tropical Africa. The seeds contain multiple forms of Bowman Birk type inhibitors. The major inhibitor HGI-III, from the native seed with 76 amino acid residues exists as a dimer. The amino acid sequence of three isoforms of Bowman-Birk inhibitor from germinated horsegram, designated as HGGI-I, HGGI II, and HGGI-III, have been obtained by sequential Edman analyses of the pyridylethylated inhibitors and peptides derived therefrom by enzymatic and chemical cleavage. The HGGIs are monomers, comprising of 66, 65, and 60 amino acid residues, respectively. HGGI-III from the germinated seed differs from the native seed inhibitor in the physiological deletion of a dodecapeptide at the amino terminus and a tetrapeptide, -SHDD, at the carboxyl terminus. The study of the state of association of HGI-III, by size-exclusion chromatography and SDS PAGE in the presence of 1 mM ZnCl2, has revealed the role of charged interactions in the monomer <--> dimer equilibria. Chemical modification studies of Lys and Arg have confirmed the role of charge interactions in the above equilibria. These results support the premise that a unique interaction, which stabilizes the dimer, is the cause of self-association in the inhibitors. This interaction in HGI-III involves the epsilon-amino group of the Lys24 (P1 residue) at the first reactive site of one monomer and the carboxyl of an Asp86 at the carboxyl terminus of the second monomer. Identification of the role of these individual amino acids in the structure and stability of the dimer was accomplished by chemical modifications, multiple sequence alignment of legume Bowman-Birk inhibitors, and homology modeling. The state of association may also influence the physiological and functional role of these inhibitors. PMID- 15123730 TI - Operator design and mechanism for CarA repressor-mediated down-regulation of the photoinducible carB operon in Myxococcus xanthus. AB - The carB operon encodes all except one of the enzymes involved in light-induced carotenogenesis in Myxococcus xanthus. Expression of its promoter (P(B)) is repressed in the dark by sequence-specific DNA binding of CarA to a palindrome (pI) located between positions -47 and -64 relative to the transcription start site. This promotes subsequent binding of CarA to additional sites that remain to be defined. CarS, produced in the light, interacts physically with CarA, abrogates CarA-DNA binding, and thereby derepresses P(B). In this study, we delineate the operator design that exists for CarA by precisely mapping out the second operator element. For this, we examined how stepwise deletions and site directed mutagenesis in the region between the palindrome and the transcription start site affect CarA binding around P(B) in vitro and expression of P(B) in vivo. These revealed the second operator element to be an imperfect interrupted palindrome (pII) spanning positions -26 to -40. In vitro assays using purified M. xanthus RNA polymerase showed that CarA abolishes P(B)-RNA polymerase binding and runoff transcription and that both were restored by CarS, thus rationalizing the observations in vivo. CarA binding to pII (after association with pI) effectively occludes RNA polymerase from P(B) and so provides the operative mechanism for the repression of the carB operon by CarA. The bipartite operator design, whereby transcription is blocked by the low affinity CarA-pII binding and is readily restored by CarS, may have evolved to match the needs for a rapid and an effective response to light. PMID- 15123731 TI - Transcriptional regulation of rat CYP2A3 by nuclear factor 1: identification of a novel NFI-A isoform, and evidence for tissue-selective interaction of NFI with the CYP2A3 promoter in vivo. AB - Rat CYP2A3 and its mouse and human orthologs are expressed preferentially in the olfactory mucosa. We found previously that an element in the proximal promoter region of CYP2A3 (the nasal predominant transcriptional activating (NPTA) element), which is similar to a nuclear factor 1 (NFI)-binding site, is critical for transcriptional activation of CYP2A3 in vitro. We proposed that this element might be important for tissue-selective CYP2A3 expression. The goals of the present study were to characterize NPTA-binding proteins and to obtain more definitive evidence for the role of NFI in the transcriptional activation of CYP2A3. The NPTA-binding proteins were isolated by DNA-affinity purification from rat olfactory mucosa. Mass spectral analysis indicated that isoforms corresponding to all four NFI genes were present in the purified NPTA-binding fraction. Further analysis of NPTA-binding proteins led to the identification of a novel NFI-A isoform, NFI-A-short, which was derived from alternative splicing of the NFI-A transcript. Transient transfection assay showed that NFI-A2, an NFI isoform previously identified in the olfactory mucosa, transactivated the CYP2A3 promoter, whereas NFI-A-short, which lacks the transactivation domain, counteracted the activation. Chromatin immunoprecipitation assays indicated that NFI proteins are associated with the CYP2A3 promoter in vivo, in rat olfactory mucosa, but essentially not in the liver where the CYP2A3 promoter is hypermethylated and CYP2A3 is not expressed. These data strongly support a role for NFI transcription factors in the transcriptional activation of CYP2A3. PMID- 15123732 TI - Chemotherapy for melanoma: the resultant of conflicting vectors. PMID- 15123733 TI - Exposure of melanoma cells to dacarbazine results in enhanced tumor growth and metastasis in vivo. AB - PURPOSE: In recent years, the incidence of cutaneous melanoma has increased more than that of any other cancer. Dacarbazine is considered the gold standard for treatment, having a response rate of 15% to 20%, but most responses are not sustained. Previously, we have shown that short exposure of primary cutaneous melanoma cells to dacarbazine resulted in the upregulation of interleukin-8 (IL 8) and vascular endothelial growth factor (VEGF). The purpose of the present study was to determine how long-term exposure of melanoma cells to dacarbazine would affect their tumorigenic and metastatic potential in vivo. MATERIALS AND METHODS: The primary cutaneous melanoma cell lines SB2 and MeWo were repeatedly exposed in vitro to increasing concentrations of dacarbazine, and dacarbazine resistant cell lines SB2-D and MeWo-D were selected and examined for their ability to grow and metastasize in nude mice. RESULTS: The dacarbazine-resistant cell lines SB2-D and MeWo-D exhibited increased tumor growth and metastatic behavior in vivo. This increase could be explained by the activation of RAF, MEK, and ERK, which led to the upregulation of IL-8 and VEGF. More IL-8, VEGF, matrix metalloproteinase-2 (MMP-2), and microvessel density (CD-31) were found in tumors produced by SB2-D and MeWo-D in vivo than in those produced by their parental counterparts. No mutations were observed in BRAF. CONCLUSION: Our results have significant clinical implications. Treatment of melanoma patients with dacarbazine could select for a more aggressive melanoma phenotype. We propose that combination treatment with anti-VEGF/IL-8 or MEK inhibitors may potentiate the therapeutic effects of dacarbazine. PMID- 15123734 TI - All systems go. AB - Reductionism has served cell biologists well in the past decades, but systems biologists are now promising to broaden the picture. The challenge will be to provide not just the lists and quantitation but real mechanistic insights. PMID- 15123735 TI - Degradative organelles containing mislocalized alpha-and beta-synuclein proliferate in presenilin-1 null neurons. AB - Presenilin-1 null mutation (PS1 -/-) in mice is associated with morphological alterations and defects in cleavage of transmembrane proteins. Here, we demonstrate that PS1 deficiency also leads to the formation of degradative vacuoles and to the aberrant translocation of presynaptic alpha- and beta synuclein proteins to these organelles in the perikarya of primary neurons, concomitant with significant increases in the levels of both synucleins. Stimulation of autophagy in control neurons produced a similar mislocalization of synucleins as genetic ablation of PS1. These effects were not the result of the loss of PS1 gamma-secretase activity; however, dysregulation of calcium channels in PS1 -/- cells may be involved. Finally, colocalization of alpha-synuclein and degradative organelles was observed in brains from patients with the Lewy body variant of AD. Thus, aberrant accumulation of alpha- and beta-synuclein in degradative organelles are novel features of PS1 -/- neurons, and similar events may promote the formation of alpha-synuclein inclusions associated with neurodegenerative diseases. PMID- 15123736 TI - ERK activation promotes neuronal degeneration predominantly through plasma membrane damage and independently of caspase-3. AB - Our recent studies have shown that extracellular-regulated protein kinase (ERK) promotes cell death in cerebellar granule neurons (CGN) cultured in low potassium. Here we report that the "death" phenotypes of CGN after potassium withdrawal are heterogeneous, allowing the distinction between plasma membrane (PM)-, DNA-, and PM/DNA-damaged populations. These damaged neurons display nuclear condensation that precedes PM or DNA damage. Inhibition of ERK activation either by U0126 or by dominant-negative mitogen-activated protein kinase/ERK kinase (MEK) overexpression results in a dramatic reduction of PM damaged neurons and nuclear condensation. In contrast, overexpression of constitutively active MEK potentiates PM damage and nuclear condensation. ERK-promoted cellular damage is independent of caspase-3. Persistent active ERK translocates to the nucleus, whereas caspase-3 remains in the cytoplasm. Antioxidants that reduced ERK activation and PM damage showed no effect on caspase-3 activation or DNA damage. These data identify ERK as an important executor of neuronal damage involving a caspase-3-independent mechanism. PMID- 15123737 TI - Proximal, selective, and dynamic interactions between integrin alphaIIbbeta3 and protein tyrosine kinases in living cells. AB - Stable platelet aggregation, adhesion, and spreading during hemostasis are promoted by outside-in alphaIIbbeta3 signals that feature rapid activation of c Src and Syk, delayed activation of FAK, and cytoskeletal reorganization. To evaluate these alphaIIbbeta3-tyrosine kinase interactions at nanometer proximity in living cells, we monitored bioluminescence resonance energy transfer between GFP and Renilla luciferase chimeras and bimolecular fluorescence complementation between YFP half-molecule chimeras. These techniques revealed that alphaIIbbeta3 interacts with c-Src at the periphery of nonadherent CHO cells. After plating cells on fibrinogen, complexes of alphaIIbbeta3-c-Src, alphaIIbbeta3-Syk, and c Src-Syk are observed in membrane ruffles and focal complexes, and the interactions involving Syk require Src activity. In contrast, FAK interacts with alphaIIbbeta3 and c-Src, but not with Syk, in focal complexes and adhesions. All of these interactions require the integrin beta3 cytoplasmic tail. Thus, alphaIIbbeta3 interacts proximally, if not directly, with tyrosine kinases in a coordinated, selective, and dynamic manner during sequential phases of alphaIIbbeta3 signaling to the actin cytoskeleton. PMID- 15123738 TI - Identification of the membrane receptor of a class XIV myosin in Toxoplasma gondii. AB - Apicomplexan parasites exhibit a unique form of substrate-dependent motility, gliding motility, which is essential during their invasion of host cells and during their spread between host cells. This process is dependent on actin filaments and myosin that are both located between the plasma membrane and two underlying membranes of the inner membrane complex. We have identified a protein complex in the apicomplexan parasite Toxoplasma gondii that contains the class XIV myosin required for gliding motility, TgMyoA, its associated light chain, TgMLC1, and two novel proteins, TgGAP45 and TgGAP50. We have localized this complex to the inner membrane complex of Toxoplasma, where it is anchored in the membrane by TgGAP50, an integral membrane glycoprotein. Assembly of the protein complex is spatially controlled and occurs in two stages. These results provide the first molecular description of an integral membrane protein as a specific receptor for a myosin motor, and further our understanding of the motile apparatus underlying gliding motility in apicomplexan parasites. PMID- 15123739 TI - Smad6/Smurf1 overexpression in cartilage delays chondrocyte hypertrophy and causes dwarfism with osteopenia. AB - Biochemical experiments have shown that Smad6 and Smad ubiquitin regulatory factor 1 (Smurf1) block the signal transduction of bone morphogenetic proteins (BMPs). However, their in vivo functions are largely unknown. Here, we generated transgenic mice overexpressing Smad6 in chondrocytes. Smad6 transgenic mice showed postnatal dwarfism with osteopenia and inhibition of Smad1/5/8 phosphorylation in chondrocytes. Endochondral ossification during development in these mice was associated with almost normal chondrocyte proliferation, significantly delayed chondrocyte hypertrophy, and thin trabecular bone. The reduced population of hypertrophic chondrocytes after birth seemed to be related to impaired bone growth and formation. Organ culture of cartilage rudiments showed that chondrocyte hypertrophy induced by BMP2 was inhibited in cartilage prepared from Smad6 transgenic mice. We then generated transgenic mice overexpressing Smurf1 in chondrocytes. Abnormalities were undetectable in Smurf1 transgenic mice. Mating Smad6 and Smurf1 transgenic mice produced double transgenic pups with more delayed endochondral ossification than Smad6 transgenic mice. These results provided evidence that Smurf1 supports Smad6 function in vivo. PMID- 15123741 TI - SLAM Family Receptors Regulate Immunity with and without SAP-related Adaptors. AB - Two papers describing mice deficient in signaling lymphocyte activation molecule and 2B4 represent the first accounts of immune phenotypes in animals lacking members of the SLAM family of receptors. The findings provide definitive evidence of the importance of SLAM-related receptors in the regulation of T cell, macrophage, and natural killer cell functions. PMID- 15123740 TI - Involvement of caspase-4 in endoplasmic reticulum stress-induced apoptosis and Abeta-induced cell death. AB - Recent studies have suggested that neuronal death in Alzheimer's disease or ischemia could arise from dysfunction of the endoplasmic reticulum (ER). Although caspase-12 has been implicated in ER stress-induced apoptosis and amyloid-beta (Abeta)-induced apoptosis in rodents, it is controversial whether similar mechanisms operate in humans. We found that human caspase-4, a member of caspase 1 subfamily that includes caspase-12, is localized to the ER membrane, and is cleaved when cells are treated with ER stress-inducing reagents, but not with other apoptotic reagents. Cleavage of caspase-4 is not affected by overexpression of Bcl-2, which prevents signal transduction on the mitochondria, suggesting that caspase-4 is primarily activated in ER stress-induced apoptosis. Furthermore, a reduction of caspase-4 expression by small interfering RNA decreases ER stress induced apoptosis in some cell lines, but not other ER stress-independent apoptosis. Caspase-4 is also cleaved by administration of Abeta, and Abeta induced apoptosis is reduced by small interfering RNAs to caspase-4. Thus, caspase-4 can function as an ER stress-specific caspase in humans, and may be involved in pathogenesis of Alzheimer's disease. PMID- 15123742 TI - Plasmodium falciparum associated with severe childhood malaria preferentially expresses PfEMP1 encoded by group A var genes. AB - Parasite-encoded variant surface antigens (VSAs) like the var gene-encoded Plasmodium falciparum erythrocyte membrane protein 1 (PfEMP1) family are responsible for antigenic variation and infected red blood cell (RBC) cytoadhesion in P. falciparum malaria. Parasites causing severe malaria in nonimmune patients tend to express a restricted subset of VSA (VSA(SM)) that differs from VSA associated with uncomplicated malaria and asymptomatic infection (VSA(UM)). We compared var gene transcription in unselected P. falciparum clone 3D7 expressing VSA(UM) to in vitro-selected sublines expressing VSA(SM) to identify PfEMP1 responsible for the VSA(SM) phenotype. Expression of VSA(SM) was accompanied by up-regulation of Group A var genes. The most prominently up regulated Group A gene (PFD1235w/MAL7P1.1) was translated into a protein expressed on the infected RBC surface. The proteins encoded by Group A var genes, such as PFD1235w/MAL7P1.1, appear to be involved in the pathogenesis of severe disease and are thus attractive candidates for a vaccine against life-threatening P. falciparum malaria. PMID- 15123743 TI - Natural killer T cells infiltrate neuroblastomas expressing the chemokine CCL2. AB - CD1d-restricted Valpha24-Jalpha18-invariant natural killer T cells (iNKTs) are potentially important in tumor immunity. However, little is known about their localization to tumors. We analyzed 98 untreated primary neuroblastomas from patients with metastatic disease (stage 4) for tumor-infiltrating iNKTs using TaqMan((R)) reverse transcription polymerase chain reaction and immunofluorescent microscopy. 52 tumors (53%) contained iNKTs, and oligonucleotide microarray analysis of the iNKT(+) and iNKT(-) tumors revealed that the former expressed higher levels of CCL2/MCP-1, CXCL12/SDF-1, CCL5/RANTES, and CCL21/SLC. Eight tested neuroblastoma cell lines secreted a range of CCL2 (0-21.6 ng/ml), little CXCL12 (95% oxygen) for 24 to 72 h, and pulmonary and hepatic expression of CYP1A and lung injury were studied. Hyperoxia caused significant increases in pulmonary and hepatic CYP1A1 activities (ethoxyresorufin O-deethylase) and mRNA levels in wild type (C57BL/6J) AHR (+/+), but not AHR (-/-) mice, suggesting that AHR-dependent mechanisms contributed to CYP1A1 induction. On the other hand, hyperoxia augmented hepatic CYP1A2 expression in both wild-type and AHR (-/-) animals, suggesting that AHR-independent mechanisms contributed to the CYP1A2 regulation by hyperoxia. AHR (-/-) mice exposed to hyperoxia were more susceptible than wild type mice to lung injury and inflammation, as indicated by significantly higher lung weight/body weight ratios, increased pulmonary edema, and enhanced neutrophil recruitment into the lungs. In conclusion, our results support the hypothesis that the hyperoxia induces CYP1A1, but not CYP1A2, expression in vivo by AHR-dependent mechanisms, a phenomenon that may mechanistically contribute to the beneficial effects of the AHR in hyperoxic lung injury. PMID- 15123766 TI - Flavonoids from artichoke (Cynara scolymus L.) up-regulate endothelial-type nitric-oxide synthase gene expression in human endothelial cells. AB - Nitric oxide (NO) produced by endothelial nitric-oxide synthase (eNOS) represents an antithrombotic and anti-atherosclerotic principle in the vasculature. Hence, an enhanced expression of eNOS in response to pharmacological interventions could provide protection against cardiovascular diseases. In EA.hy 926 cells, a cell line derived from human umbilical vein endothelial cells (HUVECs), an artichoke leaf extract (ALE) increased the activity of the human eNOS promoter (determined by luciferase reporter gene assay). An organic subfraction from ALE was more potent in this respect than the crude extract, whereas an aqueous subfraction of ALE was without effect. ALE and the organic subfraction thereof also increased eNOS mRNA expression (measured by an RNase protection assay) and eNOS protein expression (determined by Western blot) both in EA.hy 926 cells and in native HUVECs. NO production (measured by NO-ozone chemiluminescence) was increased by both extracts. In organ chamber experiments, ex vivo incubation (18 h) of rat aortic rings with the organic subfraction of ALE enhanced the NO-mediated vasodilator response to acetylcholine, indicating that the up-regulated eNOS remained functional. Caffeoylquinic acids and flavonoids are two major groups of constituents of ALE. Interestingly, the flavonoids luteolin and cynaroside increased eNOS promoter activity and eNOS mRNA expression, whereas the caffeoylquinic acids cynarin and chlorogenic acid were without effect. Thus, in addition to the lipid-lowering and antioxidant properties of artichoke, an increase in eNOS gene transcription may also contribute to its beneficial cardiovascular profile. Artichoke flavonoids are likely to represent the active ingredients mediating eNOS up-regulation. PMID- 15123767 TI - Analysis of the effects of cannabinoids on synaptic transmission between basket and Purkinje cells in the cerebellar cortex of the rat. AB - The hypothesis of the present work was that activation of CB1 cannabinoid receptors inhibits GABAergic neurotransmission between basket and Purkinje cells in the cerebellar cortex. The aim was to test this hypothesis under near physiological conditions. Action potentials of basket cells and spontaneous inhibitory postsynaptic currents (sIPSCs) in synaptically coupled Purkinje cells were recorded simultaneously in rat brain slices. The cannabinoid agonists (R) (+)-[2,3-dihydro-5-methyl-3-[(morpholinyl) methyl] pyrrolo[1,2,3-de]-1,4 benzoxazinyl]-(1-naphthalenyl)methanone mesylate (WIN 55212-2) and (-)-cis-3-[2 hydroxy-4-(1,1-dimethylheptyl)-phenyl]-trans-4-(3-hydroxy-propyl)-cyclohexanol (CP55940) decreased the amplitude of sIPSCs occurring simultaneously with basket cell action potentials and lowered the success rate of synaptic transmission. These effects were prevented by the CB1 receptor antagonist N-piperidino-5-(4 chlorophenyl)-1-(2,4-dichloro-phenyl)-4-methyl-3-pyrazole-carboxamide (SR141716). Depolarization of Purkinje cells also led to suppression of neurotransmission; prevention of this suppression by CP55940 and SR141716 indicates that endocannabinoids released from Purkinje cells were involved. WIN 55212-2 lowered the amplitude of autoreceptor currents recorded in basket cells (autoreceptor currents are due to the action of GABA released from axon terminals on GABAA autoreceptors of the same axon terminals); this is novel proof of the presynaptic action of cannabinoids. Autoreceptor current experiments also indicated that endogenous cannabinoids are not released by basket cell axon terminals. A presynaptic action is additionally supported by the observation that WIN 55212-2 lowered the frequency of miniature IPSCs recorded in the presence of tetrodotoxin and the calcium ionophore ionomycin. In conclusion, activation of CB1 receptors by exogenous cannabinoids and by endogenous cannabinoids released by Purkinje cells presynaptically inhibits GABAergic neurotransmission between basket and Purkinje cells. This was demonstrated under near-physiological conditions: transmitter release was elicited by action potentials generated by spontaneously firing intact presynaptic neurons. PMID- 15123768 TI - Bioactivation of 1,1-dichloroethylene by CYP2E1 and CYP2F2 in murine lung. AB - 1,1-Dichloroethylene (DCE) exposure evokes lung toxicity with selective damage to bronchiolar Clara cells. Recent in vitro studies have implicated CYP2E1 and CYP2F2 in the bioactivation of DCE to 2-S-glutathionyl acetate [C], a putative conjugate of DCE epoxide with glutathione. An objective of this study was to test the hypothesis that bioactivation of DCE is catalyzed by both CYP2E1 and CYP2F2 in murine lung. Western blot analysis of lung microsomal proteins from DCE treated CD-1 mice showed time-dependent loss of immunodetectable CYP2F2 and CYP2E1 protein. Dose-dependent formation of conjugate [C] was observed in the lungs of CD-1 mice treated with DCE (75-225 mg/kg), but it was not detected after pretreatment with 5-phenyl-1-pentyne (5-PIP). Treatment of mice with 5-PIP and also with diallyl sulfone (DASO2) significantly inhibited hydroxylation of p nitrophenol (PNP) and chlorzoxazone (CHZX). Incubation of recombinant CYP2F3 (a surrogate for CYP2F2) and recombinant CYP2E1 with PNP and CHZX confirmed that they are substrates for both of the recombinant enzymes. Incubation of the recombinant enzymes with DASO2 or 5-PIP significantly inhibited hydroxylation of both PNP and CHZX. Bronchiolar injury was elicited in CD-1 mice treated with DCE (75 mg/kg), but it was abrogated with 5-PIP pretreatment. Bronchiolar toxicity also was manifested in the lungs of CYP2E1-null and wild-type mice treated with DCE (75 mg/kg), but protection ensued after pretreatment with 5-PIP or DASO2. These results showed that bioactivation of DCE in murine lung occurred via the catalytic activities of both CYP2E1 and CYP2F2 and that bioactivation by these enzymes mediated the lung toxicity. PMID- 15123769 TI - IFN-gamma inhibits the proliferation of allergen-activated T lymphocytes from atopic, asthmatic patients by inducing Fas/FasL-mediated apoptosis. AB - The defect in interferon-gamma (IFN-gamma) production that results in a T helper cell type 2-dominated response may be responsible for a decrease in the apoptosis of allergen-activated T cells in asthma. We investigated the effect of recombinant IFN-gamma on proliferation, Fas/Fas ligand (FasL) expression, and apoptosis in allergen-stimulated peripheral blood mononuclear cells obtained from atopic, asthmatic patients and nonatopic, control subjects. The addition of IFN gamma at the start of cultures markedly inhibited the proliferative response to a specific allergen in cells from all asthmatic patients, whereas no change was observed in cells from nonatopic, control subjects. IFN-gamma induced an increase in the expression of Fas and FasL by allergen-stimulated CD4+ T cells from asthmatic patients and caused the apoptosis of these cells. A Fas-blocking monoclonal antibody prevented the inhibitory effect of IFN-gamma on allergen induced proliferation. These results suggest that IFN-gamma inhibits the proliferation of allergen-stimulated CD4+ T cells from atopic, asthmatic patients by inducing the surface expression of Fas and FasL, which in turn triggers their apoptotic program. The defect in IFN-gamma production involved in the allergic, immune response may therefore be responsible for a decrease in apoptosis of allergen-activated T lymphocytes in the airways of atopic, asthmatic patients. PMID- 15123770 TI - Interleukin-15 enhances human neutrophil phagocytosis by a Syk-dependent mechanism: importance of the IL-15Ralpha chain. AB - Interleukin-15 (IL-15) is a cytokine that possesses interesting, potential therapeutic properties. However, based on several parameters including activation of neutrophils, it is also recognized as a proinflammatory cytokine. The mechanisms by which IL-15 activates human neutrophil functions are not fully understood. Although these cells express a functional IL-15 receptor (IL-15R) composed of IL-15Ralpha, IL-2/15Rbeta (CD122), and gamma(c) (CD132) subunits, the role of each receptor component has not been investigated in IL-15-induced human neutrophil responses. In the present study, fluorescein-activated cell sorter analysis revealed that the ability of IL-15 to enhance neutrophil phagocytosis is not a result of increased expression of IL-15Ralpha, CD122, or CD132 on the neutrophil cell surface. Pretreatment of neutrophils with specific antibodies to IL-15Ralpha, CD122, or CD132 was found to inhibit phagocytosis of opsonized-sheep red blood cells by nearly 40%, 21%, and 27%, respectively. As expected, pretreatment of neutrophils with anti-IL-2Ralpha (CD25) had no effect. Pretreatment of cells with the Syk inhibitor piceatannol was found to significantly inhibit the ability of IL-15 to enhance phagocytosis. In addition, IL-15 was found to induce tyrosine phosphorylation of Syk that was largely inhibited by pretreating cells with piceatannol. Moreover, we found that Syk kinase is physically associated with IL-15Ralpha. We conclude that IL-15R enhances neutrophil phagocytosis by a Syk-dependent mechanism and that the IL 15Ralpha chain plays a key role in mediating this response, at least by interacting with Syk kinase. PMID- 15123771 TI - CXCR2 inhibition suppresses hemorrhage-induced priming for acute lung injury in mice. AB - Polymorphonuclear neutrophil (PMN) extravasation/sequestration in the lung and a dysregulated inflammatory response characterize the pathogenesis of acute lung injury (ALI). Previously, we have shown that hemorrhage (Hem) serves to prime PMN such that subsequent septic challenge [cecal ligation and puncture (CLP)] produces a pathological, inflammatory response and consequent lung injury in mice. Keratinocyte-derived chemokine (KC) and macrophage inflammatory protein-2 (MIP-2) are murine CXC chemokines found elevated in the lungs and plasma following Hem/CLP and have been reported by others to share a common receptor (CXCR2). Based on these data, we hypothesize that blockade of CXCR2 immediately following Hem would suppress KC and MIP-2 priming of PMN, thereby reducing the inflammatory injury observed following CLP. To assess this, Hem mice (90 min at 35+/-5 mmHg) were randomized to receive 0, 0.4, or 1 mg antileukinate (a hexapeptide inhibitor of CXCRs) in 100 microl phosphate-bufferd saline (PBS)/mouse subcutaneously, immediately following resuscitation (Ringer's lactate 4x drawn blood volume). Twenty-four hours post-Hem, mice were subjected to CLP and killed 24 h later. The results show that blockade of CXCR2 significantly (P<0.05, Tukey's test) reduced PMN influx, lung protein leak, and lung-tissue content of interleukin (IL)-6, KC, and MIP-2 and increased tissue IL-10 levels. Plasma IL-6 was significantly decreased, and IL-10 levels increased in a dose dependent manner compared with PBS-treated mice. A differential effect was observed in plasma levels of KC and MIP-2. KC showed a significant reduction at the 0.4 mg antileukinate dose. In contrast, plasma MIP-2 was significantly elevated at both doses compared with the PBS-treated controls. Together, these data demonstrate that blockade of CXCR2 signaling attenuates shock-induced priming and ALI observed following Hem and subsequent septic challenge in mice. PMID- 15123772 TI - CCL2, a product of mice early after systemic inflammatory response syndrome (SIRS), induces alternatively activated macrophages capable of impairing antibacterial resistance of SIRS mice. AB - Infection associated with systemic inflammatory response syndrome (SIRS) is a major cause of morbidity and mortality in patients with major surgery, polytrauma, and severe burn injury. In previous studies, mice with severe pancreatitis (a mouse model of SIRS, SIRS mice) have been shown to be greatly susceptible to various infections. In the present study, a mechanism involved in the impaired resistance of SIRS mice to infectious complications was investigated. Sera from SIRS mice impaired the resistance of normal mice to infectious complications induced by cecal ligation and puncture (CLP). CC chemokine ligand 2 (CCL2) was detected in sera of SIRS mice. Resident macrophages (RMphi) cultured with SIRS mouse sera converted to alternatively activated macrophages (AAMphi), which were also demonstrated in mice treated with recombinant murine CCL2. However, AAMphi were not demonstrated in mice injected with SIRS mouse sera and anti-CCL2 monoclonal antibody (mAb) in combination. Furthermore, normal mice that received SIRS mouse sera and anti-CCL2 mAb resisted CLP-induced infectious complications. These results indicate that the resistance of SIRS mice to infectious complications is impaired by AAMphi generated from RMphi in response to SIRS-associated CCL2 production. PMID- 15123773 TI - P-selectin glycoprotein ligand-1 is required for the development of cutaneous vasculitis induced by immune complex deposition. AB - Immune complex (IC)-induced tissue injury is mediated by inflammatory cell infiltration that is highly regulated by various adhesion molecules. To assess the contribution of P-selectin glycoprotein ligand-1 (PSGL-1) and selectins in the pathogenetic process, the cutaneous reverse-passive Arthus reaction was examined in mice treated with monoclonal antibodies (mAb) to PSGL-1 or P- and/or E-selectin. Edema and hemorrhage were significantly reduced in mice treated with anti-P-selectin mAb compared with control mice while they were not inhibited in mice treated with anti-E-selectin mAb. It is remarkable that blocking PSGL-1 by mAb resulted in significant, further reduction in edema and hemorrhage compared with blocking anti-P- or anti-E-selectin. However, blockade of E- and P-selectins exhibited more significant reduction relative to PSGL-1 blockade. The inhibited edema and hemorrhage paralleled reduced infiltration of neutrophils and mast cells. Reduced infiltration of neutrophils and mast cells was observed in the peritoneal Arthus reaction and was associated with the decreased production of tumor necrosis factor alpha and interleukin-6. The results of this study indicate that PSGL-1 contributes to the Arthus reaction mainly as a ligand of P-selectin and partly as a ligand of E- and/or L-selectin by regulating neutrophil and mast cell recruitment and that PSGL-1 would be a therapeutic target for human IC mediated diseases. PMID- 15123774 TI - Cross-linking of MHC class I molecules on human NK cells inhibits NK cell function, segregates MHC I from the NK cell synapse, and induces intracellular phosphotyrosines. AB - Engagement of major histocompatibility complex (MHC) class I molecules on immune cells, where they are usually highly expressed, induces signal transduction events of unclear significance. We show here that antibody-mediated cross-linking of MHC-I molecules on human natural killer (NK) cells inhibits their cytotoxic activity against tumor target cells. Inhibition by anti-MHC class I monoclonal antibody exhibits molecular specificity and is an isotype and Fc-independent process. Physical hindrance of specific molecular recognition, induction of apoptosis, or reciprocal NK cell killing, which could be induced by cross-linking of MHC I molecules, has also been ruled out as putative mechanisms of inhibition. Confocal microscopy analysis revealed that MHC class I molecules on the surface of NK cells colocalize constitutively with GM1, a marker of lipid rafts. Cross linking of MHC class I resulted in the asymmetric redistribution of GM1-enriched raft domains, which are concentrated to the immunological synapse, and MHC I molecules, which segregate to the opposite pole. Also, the cross-linking of MHC I on NK cells induced intracellular tyrosine phosphorylations. These results suggest that MHC I molecules on NK cells could transmit inhibitory signals upon engagement with putative ligands expressed on the surface of those cells that need to be protected from natural cytotoxicity. PMID- 15123775 TI - CD1d-restricted "NKT" cells and myeloid IL-12 production: an immunological crossroads leading to promotion or suppression of effective anti-tumor immune responses? AB - CD1d-restricted T cells are remarkable for their unusual ability to respond to self-antigens and to contribute to both immunostimulatory and immunosuppressive responses. Their effects in different cancer models have appeared contradictory; in some cases, they are linked to the generation of effective tumor clearance, and in others, they seem to contribute to suppression of anti-tumor responses. Recent results suggest CD1d-restricted T cells are involved in critical interactions with myeloid dendritic cells (DCs) that can affect the subsequent course of the immune response, and that factors such as the strength of the antigenic signal and the presence or absence of proinflammatory cytokines may determine the outcome of these interactions. In the presence of a strong antigenic signal, CD1d-restricted T cells induced myeloid DCs to secrete interleukin (IL)-12, and these DCs in turn activated naive T cells to secrete Th1 cytokines. When exposed to the weak antigenic stimulus of self-antigens, CD1d restricted T cells induced DCs to secrete IL-10 but not IL-12, and these DCs failed to stimulate Th1 cytokine production by naive T cells. In contrast, CD1d restricted T cells that were stimulated by self-antigens in the presence of IL-12 potently secreted interferon-gamma (IFN-gamma) and were among the first lymphocytes to become activated in vivo. Hence, CD1d-restricted T cells may promote or prevent effective anti-tumor responses that are mediated by other lymphocytic effector cells by influencing IL-12 production by myeloid DCs and by their own production of early IFN-gamma in response to IL-12. PMID- 15123776 TI - The expanded family of class II cytokines that share the IL-10 receptor-2 (IL 10R2) chain. AB - Several novel interleukin (IL)-10-related cytokines have recently been discovered. These include IL-22, IL-26, and the interferon-lambda (IFN-lambda) proteins IFN-lambda1 (IL-29), IFN-lambda2 (IL-28A), and IFN-lambda3 (IL-28B). The ligand-binding chains for IL-22, IL-26, and IFN-lambda are distinct from that used by IL-10; however, all of these cytokines use a common second chain, IL-10 receptor-2 (IL-10R2; CRF2-4), to assemble their active receptor complexes. Thus, IL-10R2 is a shared component in at least four distinct class II cytokine receptor complexes. IL-10 binds to IL-10R1; IL-22 binds to IL-22R1; IL-26 binds to IL-20R1; and IFN-lambda binds to IFN-lambdaR1 (also known as IL-28R). The binding of these ligands to their respective R1 chains induces a conformational change that enables IL-10R2 to interact with the newly formed ligand-receptor complexes. This in turn activates a signal-transduction cascade that results in rapid activation of several transcription factors, particularly signal transducer and activator of transcription (STAT)3 and to a lesser degree, STAT1. Activation by IL-10, IL-22, IL-26, or IFN-lambda can be blocked with neutralizing antibodies to the IL-10R2 chain. Although IL-10R2 is broadly expressed on a wide variety of tissues, only a subset of these tissues expresses the ligand-binding R1 chains. The receptors for these cytokines are often present on cell lines derived from various tumors, including liver, colorectal, and pancreatic carcinomas. Consequently, the receptors for these cytokines may provide novel targets for inhibiting the growth of certain types of cancer. PMID- 15123777 TI - Increased numbers of committed myeloid progenitors but not primitive hematopoietic stem/progenitors in mice lacking STAT6 expression. AB - Signal transducer and activator of transcription-6 (STAT6) plays important roles in cytokine signaling via interleukin-4 and -13 receptors (IL-4R and IL-13R). Mice in which STAT6 has been disrupted by homologous recombination show defects in T helper cell type 2 (Th2) lymphocyte production, resulting in an accumulation of Th1 cells. In addition to defects in differentiation and proliferation of T lymphocytes, STAT6-deficient mice show increased cell-cycle activation and frequency of myeloid progenitors. Although this has been shown to be mediated through Oncostatin M production by T cells, IL-4Ralpha and STAT6 have also recently been found to be enriched for expression in primitive hematopoietic stem cells (HSCs) in gene expression-profiling studies. Therefore, we have investigated whether defects in hematopoietic function in mice lacking STAT6 expression extended into the primitive hematopoietic compartments of the bone marrow. Here, we report that STAT6 deficiency increased bone marrow-committed myeloid progenitors but did not alter the number of cells enriched for HSC/multipotent progenitors, primitive cobblestone area-forming cells assayed in vitro, or bone marrow short-term or long-term repopulating cells assayed in vivo. Therefore, the requirement for STAT6 activation during hematopoiesis is limited, and primitive hematopoietic cell types are insulated against possible effects of cytokine stimulation by Th1 cells. PMID- 15123778 TI - Cytosolic phospholipase A2 is responsible for prostaglandin E2 and leukotriene B4 formation in phagocyte-like PLB-985 cells: studies of differentiated cPLA2 deficient PLB-985 cells. AB - Our previously established model of cytosolic phospholipase A(2) (cPLA(2)) deficient, differentiated PLB-985 cells (PLB-D cells) was used to determine the physiological role of cPLA(2) in eicosanoid production. Parent PLB-985 (PLB) cells and PLB-D cells were differentiated toward the monocyte or granulocyte lineages using 5 x 10(-)(8) M 1,25 dihydroxyvitamin D(3) or 1.25% dimethyl sulfoxide, respectively. Parent monocyte- or granulocyte-like PLB cells released prostaglandin E(2) (PGE(2)) when stimulated by ionomycin, A23187, opsonized zymosan, phorbol 12-myristate 13-acetate, or formyl-Met-Leu-Phe (fMLP), and monocyte- or granulocyte-like PLB-D cells did not release PGE(2) with any of the agonists. The kinetics of cPLA(2) translocation to nuclear fractions in monocyte like PLB cells stimulated with fMLP or ionomycin was in correlation with the kinetics of PGE(2) production. Granulocyte-like PLB cells, but not granulocyte like PLB-D cells, secreted leukotriene B(4) (LTB(4)) after stimulation with ionomycin or A23187. Preincubation of monocyte-like parent PLB cells with 100 ng/ml lipopolysaccharide (LPS) for 16 h enhanced stimulated PGE(2) production, which is in correlation with the increased levels of cPLA(2) detected in these cells. LPS preincubation was less potent in increasing PGE(2) and LTB(4) secretion and did not affect cPLA(2) expression in granulocyte-like PLB cells, which may be a result of their lower levels of surface LPS receptor expression. LPS had no effect on monocyte- or granulocyte-like PLB-D cells. The lack of eicosanoid formation in stimulated, differentiated cPLA(2)-deficient PLB cells indicates that cPLA(2) contributes to stimulated eicosanoid formation in monocyte and granulocyte-like PLB cells. PMID- 15123780 TI - Identification of genes differentially expressed in testes containing carcinoma in situ. AB - Virtually all testicular germ cell tumours originate from a common precursor, the carcinoma in situ (CIS) cell. The precise nature of the molecular mechanisms leading to CIS remains largely unknown. We performed the first systematic analysis of gene expression in testis with CIS compared to normal testis by the differential display (DDRT-PCR) method, with subsequent analysis by RT-PCR and in situ hybridization (ISH). In tissue containing CIS we identified overexpression of 28 mRNA, some previously reported in CIS and a number of genes not previously described in germ cell neoplasia, including the novel expressed sequence tag (EST) OIC1 (Overexpressed In CIS). The genes could be grouped functionally into genes involved in cell growth, proliferation, differentiation, immunological response, and genes with unknown biological function. Examples of overexpressed genes are SFRP1 that is involved in Wnt signalling and IGFBP6, which is of importance for fetal growth and inhibits cell growth through insulin-like growth factor-II. ISH analysis showed that both mRNA were localized to CIS cells. The results of our search for differentially expressed genes in CIS demonstrated a number of genes linked to testicular development (e.g. DCN, IGFBP6, SFRP1, SALL1), supporting our hypothesis that the origin of CIS is probably associated with disturbances of the fetal development of the testis. PMID- 15123779 TI - Regulation of interleukin-12 gene expression and its anti-tumor activities by prostaglandin E2 derived from mammary carcinomas. AB - Interleukin-12 (IL-12)-mediated immune responses are critical for the control of malignant development. Tumors can actively resist detrimental immunity of the host via many routes. Prostaglandin E2 (PGE2) is one of the major immune suppressive factors derived from many types of tumors. Here, we show that systemic administration of recombinant IL-12 could therapeutically control the growth of aggressive TS/A and 4T1 mouse mammary carcinomas. However, PGE2 produced by tumors potently inhibits the production of endogenous IL-12 at the level of protein secretion, mRNA synthesis, and transcription of the constituent p40 and p35 genes. The inhibition can be reversed by NS-398, a selective inhibitor of the enzymatic activity of cyclooxygenase 2 in PGE2 synthesis. Moreover, PGE2-mediated inhibition of IL-12 production requires the functional cooperation of AP-1 and AP-1 strongly suppresses IL-12 p40 transcription. Blocking PGE2 production in vivo results in a marked reduction in lung metastasis of 4T1 tumors, accompanied by enhanced ability of peritoneal macrophages to produce IL-12 and spleen lymphocytes to produce interferon-gamma. This study contributes to the elucidation of the molecular mechanisms underlying the interaction between a progressive malignancy and the immune defense apparatus. PMID- 15123781 TI - Biomarkers of genotoxicity and other end-points in an integrated approach to environmental risk assessment. AB - Risk is defined as the probability of a given toxicological hazard resulting in actual biological harm. This involves some form of mathematical relationship between exposure and toxic effects. Simplified models based on laboratory testing in surrogate species neglect potentially important factors in real life situations. Our own approach to the study of atmospheric and edaphic pollution, focused on realism, includes the use of sentinel species (animals as prospectors and integrators of information, along both the spatial and the temporal axes) and selected biomarkers. We aim to: (i) consider pollution as a complex mixture; (ii) take into account homeostasis of the environment and of living organisms; (iii) be realistic (all data obtained in the field; calculations based on actual effects; exposure measured as internal dose). The proposed test battery divides toxicological information into four blocks: systemic effects (serum biochemistry and histopathology in wild wood mice), reproduction (epididymis cell count in mice, malformations in amphibian larvae), genotoxicity (Comet test in mice and earthworms) and population effects (abundance and diversity in arthropods). Each block is represented by the sum of the results of the tests performed within the block (presented as a severity score from 0 to 3). A final value is obtained to represent the integrated toxicological harm (ITH) occurring at a given location. To assess exposure, taking into account bioavailability, we propose (i) for soil contamination studies, measuring EROD activity in liver; (ii) for atmospheric pollution, the gaseous fraction is taken from immission gases analysis, while the solid fraction is assessed through levels of metals in sentinel organisms, the values of both fractions then being combined. Finally, a regression line is established for exposure versus ITH in four to five locations with decreasing exposure levels, ranging from the immediate neighbourhood of the pollution focus to controls, following the main dissemination line. In this model we may interpolate new exposure data to find the corresponding predicted ITH. Such a prediction may be directly interpreted as a form of risk assessment or, alternatively, these pairs (toxicological harm/exposure) could then be related to a conventional scale of ecotoxicological risk. PMID- 15123782 TI - Endogenous DNA damage in humans: a review of quantitative data. AB - DNA damage plays a major role in mutagenesis, carcinogenesis and ageing. The vast majority of mutations in human tissues are certainly of endogenous origin. A thorough knowledge of the types and prevalence of endogenous DNA damage is thus essential for an understanding of the interactions of endogenous processes with exogenous agents and the influence of damage of endogenous origin on the induction of cancer and other diseases. In particular, this seems important in risk evaluation concerning exogenous agents that also occur endogenously or that, although chemically different from endogenous ones, generate the same DNA adducts. This knowledge may also be crucial to the development of rational chemopreventive strategies. A list of endogenous DNA-damaging agents, processes and DNA adduct levels is presented. For the sake of comparison, DNA adduct levels are expressed in a standardized way, including the number of adducts per 10(6) nt. This list comprises numerous reactive oxygen species and products generated as a consequence (e.g. lipid peroxides), endogenous reactive chemicals (e.g. aldehydes and S-adenosylmethionine), and chemical DNA instability (e.g. depurination). The respective roles of endogenous versus exogenous DNA damage in carcinogenesis are discussed. PMID- 15123783 TI - Genotoxicity of tamoxifen citrate and 4-nitroquinoline-1-oxide in the wing spot test of Drosophila melanogaster. AB - Tamoxifen (TAM) is an anti-oestrogen used for treatment and prevention of human breast cancer, but it is also related to human endometrial and uterine cancer. The wing spot test in Drosophila melanogaster was employed to determine the genotoxic effects of TAM and 4-nitroquinoline-1-oxide (4-NQO), a carcinogen that produces adducts similar to TAM-DNA adducts detected in rodent liver and human liver microsomes. As Drosophila spp. have no oestrogen receptor, no effects can result in binding of TAM to a receptor. Chronic treatments with TAM citrate were performed with 3-day-old larvae of the standard (ST) and high bioactivation (HB) crosses of the wing spot test at concentrations of 0.66, 1.66 and 3.33 mM. In addition, the carcinogen 4-NQO was administered at 2.5 and 5.0 mM. Somatic spots on normal wings from marker-heterozygous flies and on serrate wings from balancer heterozygous flies were scored to determine mutation and recombination events in somatic cells for each compound. The results showed genotoxic effects of TAM at 1.66 and 3.33 mM in the ST cross only and without a clear dose-response effect. This suggests a weak genotoxicity of this anti-oestrogen. The negative results obtained with TAM in the HB cross may indicate efficient detoxification of the compound by the increased xenobiotic metabolism present in this cross. As reported before, 4-NQO showed genotoxic effects in the ST cross with a clear dose response effect. For the first time, we report enhanced effects of this compound in the HB cross. It is concluded that the genotoxicity of TAM in the Drosophila wing spot test is different from that of 4-NQO. PMID- 15123784 TI - Profiles of gene expression changes in L5178Y mouse lymphoma cells treated with methyl methanesulfonate and sodium chloride. AB - Treatment of cells with genotoxic chemicals is expected to set into motion a series of events including gene expression changes to cope with the damage. We have investigated gene expression changes in L5178Y TK(+/-) mouse lymphoma cells in culture following treatment with methyl methanesulfonate (MMS), a direct acting genotoxin, and sodium chloride (NaCl), which induces mutations in these cells through indirect mechanisms at high concentrations. The mouse lymphoma cells were treated for 4 or 24 h and the cells were harvested for RNA isolation at the end of the treatment. Analysis of the transcriptome was performed using Clontech Mouse 1.2K cDNA microarrays (1185 genes) and hybridized using 32P labeled cDNA. The microwell methodology was used to quantify the mutagenic response. Of the genes examined, MMS altered the expression (1.5-fold or more) of only five (four at 4 h and one after 24 h treatment). NaCl altered two genes after 4 h treatment, but after 24 h it altered 19 genes (13 down- and six up regulated). Both compounds altered the expression of several genes associated with apoptosis and NaCl altered genes involved in DNA damage/response and GTP related proteins. This, along with other data, indicates that the widely used L5178Y TK(+/-) mouse lymphoma cells in culture are relatively recalcitrant in terms of modulating gene expression to deal with genotoxic insult. PMID- 15123785 TI - Evaluation of the mutagenic and cytotoxic effects of mercurous chloride by the micronuclei technique in golden Syrian hamsters. AB - The aims of this study were to evaluate the mutagenic and cytotoxic activity of mercurous chloride by the micronucleus technique in vivo on the bone marrow of golden Syrian hamsters after a single i.p. drug administration. Forty male golden Syrian hamsters were classified into eight groups: negative control, positive control and six groups treated with different doses of mercurous chloride (1.25, 2.5, 5, 10, 20 and 40 mg/kg). The negative control was injected with physiological saline i.p. and the positive control with cyclophosphamide at a dose of 80 mg/kg i.p. With respect to mutagenic effect, the average number of micronucleated polychromatic erythrocytes (MPE) in hamsters treated with different doses of mercurous chloride was not significant compared with the negative control. With respect to cytotoxic effect, the average polychromatic erythrocyte/red blood cell ratio showed a significant decrease when the doses were higher than the 2.5 mg/kg dose compared with the negative control. In conclusion, this preliminary study shows a cytotoxic effect but not a mutagenic effect of calomel in vivo at one time point (24 h). PMID- 15123786 TI - In vivo kinetics of micronuclei induction by bifunctional alkylating antineoplastics. AB - The aim of the present study was to determine in vivo the kinetics of micronucleated polychromatic erythrocyte (MN-PCE) induction in mice, as an approach for studying the mechanism of micronuclei induction by mitomycin C, cis diamine dichloroplatinum, busulfan and bis-chloroethylnitrosourea, bifuctional alkylating antineoplastic agents having different patterns of crosslink induction. The kinetics of MN-PCE induction was established by scoring the frequency of MN-PCE in 2000 PCE in peripheral blood, for periods of 8 or 10 h after acute treatment and up to 80 h, with different doses of the agent. The kinetics of MN-PCE induction and particularly the times of maximal induction by different bifunctional alkylating agents were compared with the kinetics previously obtained for ethylnitrosourea, methylnitrosourea and 6-mercaptopurine, agents that cause MN-PCE mainly in the first, second and third divisions after exposure, respectively. The results obtained in the present study allow us to conclude that: (i) bifunctional alkylating agents have very different efficiencies of genotoxic and cytotoxic action; (ii) all assayed bifunctional alkylating agents induced micronuclei during the first cell division, owing to the mistaken repair of primary lesions, e.g. excision; (iii) busulfan and bis chloroethylnitrosourea showed an additional late mechanism of micronuclei induction, which is expressed at the third division and seems to be related to the mismatch repair process. PMID- 15123787 TI - In vivo mutagenicity and mutation spectrum in the bone marrow and testes of B6C3F1 lacI transgenic mice following inhalation exposure to ethylene oxide. AB - The lacI mutant frequency and mutation spectrum were determined in the bone marrow and testes of B6C3F1 lacI transgenic mice exposed by inhalation to ethylene oxide (EO). Groups of male transgenic lacI B6C3F1 mice were exposed to 0, 25, 50, 100 or 200 p.p.m. EO for up to 48 weeks (6 h/day, 5 days/week) and were killed at 12, 24 or 48 weeks of EO exposure for determination of lacI mutant frequency. In the bone marrow, the lacI mutant frequency was significantly increased at the two highest exposure levels (100 and 200 p.p.m.) and at the 48 week exposure time point. The shape of the exposure-response curve for lacI mutant frequency in the bone marrow was non-linear. DNA sequence analysis of the bone marrow mutation spectrum revealed that only AT-->TA transversions occurred at an increased frequency in EO-exposed mice: 25.4% in EO-exposed mice for 48 weeks (200 p.p.m.) compared with 1.4% in air controls. In testes, the lacI mutant frequency was increased at a single exposure level of 200 p.p.m. for 24 weeks. At 48 weeks, the lacI mutant frequency in testes was significantly increased to an equal degree at 25, 50 and 100 p.p.m. EO but not at 200 p.p.m. Analysis of the testes mutation spectrum in air control mice and in mice exposed to 200 p.p.m. EO for 48 weeks revealed that no single mutational type occurred at an increased frequency. In the testes, there was a small increase across all mutational types that was sufficient to increase the overall lacI mutation frequency although not significant individually. The mutation spectrum in testes of EO-exposed mice also revealed that the increased lacI mutant frequency observed at 25 or 50 p.p.m. EO was not due to an increase in mutant siblings (clonality). These data demonstrate that inhalation exposure to EO for up to 48 weeks produces distinct mutagenic responses in bone marrow and testes. PMID- 15123788 TI - Effect of sodium arsenite on peripheral lymphocytes in vitro: individual susceptibility among a population exposed to arsenic through the drinking water. AB - Arsenic (As) contamination in ground water has affected more than 19 countries. Approximately 36 million people in the Bengal delta alone are exposed to this toxicant via drinking water (>50 microg/l) and are at potential health risk. Chronic ingestion of As via drinking water is associated with occurrence of skin lesions, cancer and other arsenic-induced diseases in West Bengal, India. An in vitro cytogenetic study was performed utilizing chromosomal aberrations (CA) in lymphocytes treated with sodium arsenite (0-5 microM) in six symptomatic (having arsenic-related skin lesions) individuals, six age- and sex-matched As-exposed asymptomatic (no arsenic-related skin lesions) individuals and six control individuals with similar socio-economic status residing in non-affected districts of West Bengal with no evidence of As exposure. The mean As content in nails and hair was 9.61 and 5.23 microg/g in symptomatic, 3.48 and 2.17 microg/g in asymptomatic and 0.42 and 0.33 microg/g in the control individuals, respectively. The main aim of our study was to determine whether genotoxic effects differed in the lymphocytes of the control (no exposure to arsenic), asymptomatic and symptomatic individuals after in vitro treatment with sodium arsenite. Although both the exposed groups had chronic exposure to As through the drinking water, individuals with skin lesions accumulated more As in their nails and hair and excreted less in urine (127.80 versus 164.15 microg/l). The results show that sodium arsenite induced a significantly higher percentage of aberrant cells in the lymphocytes of control individuals than in the lymphocytes of both the exposed groups. Within the two exposed groups As induced higher incidences of CA in the symptomatic than the asymptomatic individuals. These results suggest that asymptomatic individuals have relatively lower sensitivity and susceptibility to induction of genetic damage by As compared with the symptomatic individuals. PMID- 15123789 TI - Effects of vitamin A on doxorubicin-induced chromosomal aberrations in bone marrow cells of rats. AB - The present study was carried out to evaluate the role of vitamin A (VA) on the induction of chromosomal aberrations (CA) in rat bone marrow cells and to investigate its modulating effect on chromosomal damage induced by doxorubicin (DXR). Wistar rats were treated with VA (7.5, 15 and 30 microg/kg body wt) once a day for 2 days by gavage before injecting DXR (90 mg/kg body wt). Rats in the control group were treated with corresponding doses of water and olive oil. Animals treated with the medium dose of VA (15 microg/kg body wt) plus single dose of DXR presented a statistically significant reduction in total number of CA and in number of abnormal metaphases (P < 0.05). However, when compared with control and DXR groups, the low and high VA doses (7.5 and 30 microg/kg body wt) were found to be less efficient than the medium dose VA (15 microg/kg body wt) in terms of parameters analyzed. Furthermore, the high dose of VA group (30 microg/kg body wt) was found to be clastogenic (P < 0.05). This study concludes that the protective effect of VA against chromosome damage is dose dependent. PMID- 15123790 TI - Characterization of the hamster FancG/Xrcc9 gene and mutations in CHO UV40 and NM3. AB - The human FANCG/XRCC9 gene, which is defective in Fanconi anemia complementation group G (FA-G) cells, was first cloned by genetic complementation of the mitomycin C (MMC) sensitivity of CHO mutant UV40. The CHO NM3 mutant was subsequently assigned to the same complementation group. The parental AA8 CHO cells are hemizygous at the FancG locus, and we identified frameshift mutations that result in N-terminal truncations of the protein in both UV40 and NM3. Hypersensitivity to DNA cross-linking agents, such as MMC, typically characterizes FA cells. By introducing the native CHO FancG gene into mutant NM3, we demonstrate that hamster FancG fully corrects the 3-fold sensitivity to methyl methanesulfonate (MMS) as well as the 10-fold sensitivity to MMC, whereas resistance to ionizing radiation did not increase appreciably. In contrast, hamster cDNA transformants showed incomplete correction for both MMC and MMS sensitivity. The constitutively expressed FancG protein is present in the cytoplasmic, nuclear and chromatin fractions. FancG protein levels and subcellular localization do not change appreciably as a function of cell cycle position. Our results are consistent with roles of FancG in both the nuclear and cytoplasmic compartments to maintain genomic stability in response to various genotoxic agents. PMID- 15123791 TI - Comparative study in the Ames test of benzo[a]pyrene and 2-aminoanthracene metabolic activation using rat hepatic S9 and hepatocytes following in vivo or in vitro induction. AB - We studied the replacement of hepatic S9 with in vivo and in vitro induced hepatocytes as a metabolic activation system with the aim of broadening the possibilities of mutagenic assays. Rats were pretreated with beta-naphthoflavone (BNF), phenobarbital (PB), 3-methylcholanthrene (MC) and a combination of BNF and PB (BNF + PB). Mutagenic activation of benzo[a]pyrene (BP) and 2-aminoanthracene (2AA) by hepatic S9 and hepatocytes was determined in the Ames test. Primary rat hepatocytes were used for in vitro induction and were used as the activating system in the Ames test. In vivo BNF treatment greatly increased the metabolic activation capacity of hepatic S9 and hepatocytes towards BP. With regard to 2AA activation, S9 and hepatocytes showed different BNF induction profiles. PB treatment reduced the mutagenicity of both compounds. Although ethoxyresorufin O dealkylase (EROD) activity of S9 from BNF + PB-treated animals was almost 30-fold greater than the control, its effectiveness in activation of 2AA was below the control level. A large part of the EROD activity of control cells was lost during culture, together with the ability to activate 2AA, however, 72 h of MC induction increased EROD activity to 200-fold of the control, which corresponds to 28% of that of in vivo induced hepatocytes. The mutagenic potential of BP activated by in vitro induced hepatocytes was 10-fold above the control, which is 47% of the mutagenicity detected following in vivo induction. In vitro induced hepatocytes increased 2AA mutagenicity to 14.6-fold over the control, which corresponds to 68% of in vivo induction. Our results suggest that primary culture of hepatocytes provides a useful model for the study of the role of metabolic activation processes concerning enzyme activity of cytochromes P450 and other metabolic enzymes and induction profiles of different inducers. PMID- 15123792 TI - Production of alpha-1,3-galactosyltransferase null pigs by means of nuclear transfer with fibroblasts bearing loss of heterozygosity mutations. AB - Hyperacute rejection of porcine organs by old world primate recipients is mediated through preformed antibodies against galactosyl-alpha-1,3-galactose (Galalpha-1,3-Gal) epitopes expressed on the pig cell surface. Previously, we generated inbred miniature swine with a null allele of the alpha-1,3 galactosyltransferase locus (GGTA1) by nuclear transfer (NT) with gene-targeted fibroblasts. To expedite the generation of GGTA1 null pigs, we selected spontaneous null mutant cells from fibroblast cultures of heterozygous animals for use in another round of NT. An unexpectedly high rate of spontaneous loss of GGTA1 function was observed, with the vast majority of null cells resulting from loss of the WT allele. Healthy piglets, hemizygous and homozygous for the gene targeted allele, were produced by NT by using fibroblasts that had undergone deletional and crossover/gene conversion events, respectively. Aside from loss of Galalpha-1,3-Gal epitopes, there were no obvious phenotypic differences between these null piglets and WT piglets from the same inbred lines. In fact, congenital abnormalities observed in the heterozygous NT animals did not reappear in the serially produced null animals. PMID- 15123793 TI - The DNA-unwinding mechanism of the ring helicase of bacteriophage T7. AB - Helicases are motor proteins that use the chemical energy of NTP hydrolysis to drive mechanical processes such as translocation and nucleic acid strand separation. Bacteriophage T7 helicase functions as a hexameric ring to drive the replication complex by separating the DNA strands during genome replication. Our studies show that T7 helicase unwinds DNA with a low processivity, and the results indicate that the low processivity is due to ring opening and helicase dissociating from the DNA during unwinding. We have measured the single-turnover kinetics of DNA unwinding and globally fit the data to a modified stepping model to obtain the unwinding parameters. The comparison of the unwinding properties of T7 helicase with its translocation properties on single-stranded (ss)DNA has provided insights into the mechanism of strand separation that is likely to be general for ring helicases. T7 helicase unwinds DNA with a rate of 15 bp/s, which is 9-fold slower than the translocation speed along ssDNA. T7 helicase is therefore primarily an ssDNA translocase that does not directly destabilize duplex DNA. We propose that T7 helicase achieves DNA unwinding by its ability to bind ssDNA because it translocates unidirectionally, excluding the complementary strand from its central channel. The results also imply that T7 helicase by itself is not an efficient helicase and most likely becomes proficient at unwinding when it is engaged in a replication complex. PMID- 15123794 TI - Chlamydial histone-DNA interactions are disrupted by a metabolite in the methylerythritol phosphate pathway of isoprenoid biosynthesis. AB - The chlamydial developmental cycle is characterized by an intracellular replicative form, termed the reticulate body, and an extracellular form called the elementary body. Elementary bodies are characterized by a condensed chromatin, which is maintained by a histone H1-like protein, Hc1. Differentiation of elementary bodies to reticulate bodies is accompanied by dispersal of the chromatin as chlamydiae become transcriptionally active, although the mechanisms of Hc1 release from DNA have remained unknown. Dissociation of the nucleoid requires chlamydial transcription and translation with negligible loss of Hc1. A genetic screen was therefore designed to identify chlamydial genes rescuing Escherichia coli from the lethal effects of Hc1 overexpression. CT804, a gene homologous to ispE, which encodes an intermediate enzyme of the non-mevalonate methylerythritol phosphate (MEP) pathway of isoprenoid biosynthesis, was selected. E. coli coexpressing CT804 and Hc1 grew normally, although they expressed Hc1 to a level equivalent to that which condensed the chromatin of parent Hc1-expressing controls. Inhibition of the MEP pathway with fosmidomycin abolished IspE rescue of Hc1-expressing E. coli. Deproteinated extract from IspE expressing bacteria caused dispersal of purified chlamydial nucleoids, suggesting that chlamydial histone-DNA interactions are disrupted by a small metabolite within the MEP pathway rather than by direct action of IspE. By partial reconstruction of the MEP pathway, we determined that 2-C-methylerythritol 2,4 cyclodiphosphate dissociated Hc1 from chlamydial chromatin. These results suggest that chlamydial histone-DNA interactions are disrupted upon germination by a small metabolite in the MEP pathway of isoprenoid biosynthesis. PMID- 15123795 TI - Identification of hepoxilin A3 in inflammatory events: a required role in neutrophil migration across intestinal epithelia. AB - The mechanism by which neutrophils [polymorphonuclear leukocyte (PMNs)] are stimulated to move across epithelial barriers at mucosal surfaces has been basically unknown in biology. IL-8 has been shown to stimulate PMNs to leave the bloodstream at a local site of mucosal inflammation, but the chemical gradient used by PMNs to move between adjacent epithelial cells and traverse the tight junction at the apical neck of these mucosal barriers has eluded identification. Our studies not only identify this factor, previously termed pathogen-elicited epithelial chemoattractant, as the eicosanoid hepoxilin A(3) (hepA(3)) but also demonstrate that it is a key factor promoting the final step in PMN recruitment to sites of mucosal inflammation. We show that hepA(3) is synthesized by epithelial cells and secreted from their apical surface in response to conditions that stimulate inflammatory events. Our data further establish that hepA(3) acts to draw PMNs, via the establishment of a gradient across the epithelial tight junction complex. The functional significance of hepA(3) to target PMNs to the lumen of the gut at sites of inflammation was demonstrated by the finding that disruption of the 12-lipoxygenase pathway (required for hepA(3) production) could dramatically reduce PMN-mediated tissue trauma, demonstrating that hepA(3) is a key regulator of mucosal inflammation. PMID- 15123796 TI - Urinary concentrating defect in mice with selective deletion of phloretin sensitive urea transporters in the renal collecting duct. AB - To investigate the role of inner medullary collecting duct (IMCD) urea transporters in the renal concentrating mechanism, we deleted 3 kb of the UT-A urea transporter gene containing a single 140-bp exon (exon 10). Deletion of this segment selectively disrupted expression of the two known IMCD isoforms of UT-A, namely UT-A1 and UT-A3, producing UT-A1/3(-/-) mice. In isolated perfused IMCDs from UT-A1/3(-/-) mice, there was a complete absence of phloretin-sensitive or vasopressin-stimulated urea transport. On a normal protein intake (20% protein diet), UT-A1/3(-/-) mice had significantly greater fluid consumption and urine flow and a reduced maximal urinary osmolality relative to wild-type controls. These differences in urinary concentrating capacity were nearly eliminated when urea excretion was decreased by dietary protein restriction (4% by weight), consistent with the 1958 Berliner hypothesis stating that the chief role of IMCD urea transport in the concentrating mechanism is the prevention of urea-induced osmotic diuresis. Analysis of inner medullary tissue after water restriction revealed marked depletion of urea in UT-A1/3(-/-) mice, confirming the concept that phloretin-sensitive IMCD urea transporters play a central role in medullary urea accumulation. However, there were no significant differences in mean inner medullary Na(+) or Cl(-) concentrations between UT-A1/3(-/-) mice and wild-type controls, indicating that the processes that concentrate NaCl were intact. Thus, these results do not corroborate the predictions of passive medullary concentrating models stating that NaCl accumulation in the inner medulla depends on rapid vasopressin-regulated urea transport across the IMCD epithelium. PMID- 15123797 TI - A parieto-frontal network for visual numerical information in the monkey. AB - Recent electrophysiological studies in monkeys have implicated the prefrontal cortex (PFC) and posterior parietal cortex (PPC) in numerical judgments. The functional organization and respective contributions of these (and other) cortical areas, however, are unknown; their neural activity during numerical judgments has not been directly compared. We surveyed activity in the PPC and the anterior inferior temporal cortex while monkeys performed a visual numerosity judgment task and compared it with a population of PFC neurons. In the PPC, the proportion of numerosity-selective neurons was highest in the fundus of the intraparietal sulcus; only few numerosity-selective neurons were found in other PPC areas or the anterior inferior temporal cortex. Further, neurons in the fundus of the intraparietal sulcus responded and conveyed numerosity earlier than PFC neurons, suggesting that numerosity information flows from the PPC to the lateral PFC. This finding suggests a parieto-frontal network for numerosity in monkeys and establishes homologies between the monkey and human brain. PMID- 15123798 TI - Carbon nutrition of Escherichia coli in the mouse intestine. AB - Whole-genome expression profiling revealed Escherichia coli MG1655 genes induced by growth on mucus, conditions designed to mimic nutrient availability in the mammalian intestine. Most were nutritional genes corresponding to catabolic pathways for nutrients found in mucus. We knocked out several pathways and tested the relative fitness of the mutants for colonization of the mouse intestine in competition with their wild-type parent. We found that only mutations in sugar pathways affected colonization, not phospholipid and amino acid catabolism, not gluconeogenesis, not the tricarboxylic acid cycle, and not the pentose phosphate pathway. Gluconate appeared to be a major carbon source used by E. coli MG1655 to colonize, having an impact on both the initiation and maintenance stages. N acetylglucosamine and N-acetylneuraminic acid appeared to be involved in initiation, but not maintenance. Glucuronate, mannose, fucose, and ribose appeared to be involved in maintenance, but not initiation. The in vitro order of preference for these seven sugars paralleled the relative impact of the corresponding metabolic lesions on colonization: gluconate > N-acetylglucosamine > N-acetylneuraminic acid = glucuronate > mannose > fucose > ribose. The results of this systematic analysis of nutrients used by E. coli MG1655 to colonize the mouse intestine are intriguing in light of the nutrient-niche hypothesis, which states that the ecological niches within the intestine are defined by nutrient availability. Because humans are presumably colonized with different commensal strains, differences in nutrient availability may provide an open niche for infecting E. coli pathogens in some individuals and a barrier to infection in others. PMID- 15123799 TI - Impaired spontaneous anthropomorphizing despite intact perception and social knowledge. AB - Humans spontaneously imbue the world with social meaning: we see not only emotions and intentional behaviors in humans and other animals, but also anger in the movements of thunderstorms and willful sabotage in crashing computers. Converging evidence supports a role for the amygdala, a collection of nuclei in the temporal lobe, in processing emotionally and socially relevant information. Here, we report that a patient with bilateral amygdala damage described a film of animated shapes (normally seen as full of social content) in entirely asocial, geometric terms, despite otherwise normal visual perception. Control tasks showed that the impairment did not result from a global inability to describe social stimuli or a bias in language use, nor was a similar impairment observed in eight comparison subjects with damage to orbitofrontal cortex. This finding extends the role of the amygdala to the social attributions we make even to stimuli that are not explicitly social and, in so doing, suggests that the human capacity for anthropomorphizing draws on some of the same neural systems as do basic emotional responses. PMID- 15123800 TI - Short amino acid stretches can mediate amyloid formation in globular proteins: the Src homology 3 (SH3) case. AB - Protein misfolding and deposition underlie an increasing number of debilitating human disorders. We have shown that model proteins unrelated to disease, such as the Src homology 3 (SH3) domain of the p58alpha subunit of bovine phosphatidyl inositol-3'-kinase (PI3-SH3), can be converted in vitro into assemblies with structural and cytotoxic properties similar to those of pathological aggregates. By contrast, homologous proteins, such as alpha-spectrin-SH3, lack the capability of forming amyloid fibrils at a measurable rate under any of the conditions we have so far examined. However, transplanting a small sequence stretch (6 aa) from PI3-SH3 to alpha-spectrin-SH3, comprising residues of the diverging turn and adjacent RT loop, creates an amyloidogenic protein closely similar in its behavior to the original PI3-SH3. Analysis of specific PI3-SH3 mutants further confirms the involvement of this region in conferring amyloidogenic properties to this domain. Moreover, the inclusion in this stretch of two consensus residues favored in SH3 sequences substantially inhibits aggregation. These findings show that short specific amino acid stretches can act as mediators or facilitators in the incorporation of globular proteins into amyloid structures, and they support the suggestion that natural protein sequences have evolved in part to code for structural characteristics other than those included in the native fold, such as avoidance of aggregation. PMID- 15123801 TI - The C-terminal domain of DNA gyrase A adopts a DNA-bending beta-pinwheel fold. AB - DNA gyrase is unique among enzymes for its ability to actively introduce negative supercoils into DNA. This function is mediated in part by the C-terminal domain of its A subunit (GyrA CTD). Here, we report the crystal structure of this approximately 35-kDa domain determined to 1.75-A resolution. The GyrA CTD unexpectedly adopts an unusual fold, which we term a beta-pinwheel, that is globally reminiscent of a beta-propeller but is built of blades with a previously unobserved topology. A large, conserved basic patch on the outer edge of this domain suggests a likely site for binding and bending DNA; fluorescence resonance energy transfer-based assays show that the GyrA CTD is capable of bending DNA by > or =180 degrees over a 40-bp region. Surprisingly, we find that the CTD of the topoisomerase IV A subunit, which shares limited sequence homology with the GyrA CTD, also bends DNA. Together, these data provide a physical explanation for the ability of DNA gyrase to constrain a positive superhelical DNA wrap, and also suggest that the particular substrate preferences of topoisomerase IV might be dictated in part by the function of this domain. PMID- 15123802 TI - The structure of a thermophilic archaeal virus shows a double-stranded DNA viral capsid type that spans all domains of life. AB - Of the three domains of life (Eukarya, Bacteria, and Archaea), the least understood is Archaea and its associated viruses. Many Archaea are extremophiles, with species that are capable of growth at some of the highest temperatures and extremes of pH of all known organisms. Phylogenetic rRNA-encoding DNA analysis places many of the hyperthermophilic Archaea (species with an optimum growth > or = 80 degrees C) at the base of the universal tree of life, suggesting that thermophiles were among the first forms of life on earth. Very few viruses have been identified from Archaea as compared to Bacteria and Eukarya. We report here the structure of a hyperthermophilic virus isolated from an archaeal host found in hot springs in Yellowstone National Park. The sequence of the circular double stranded DNA viral genome shows that it shares little similarity to other known genes in viruses or other organisms. By comparing the tertiary and quaternary structures of the coat protein of this virus with those of a bacterial and an animal virus, we find conformational relationships among all three, suggesting that some viruses may have a common ancestor that precedes the division into three domains of life >3 billion years ago. PMID- 15123803 TI - Distinct localization of histone H3 acetylation and H3-K4 methylation to the transcription start sites in the human genome. AB - Almost 1-2% of the human genome is located within 500 bp of either side of a transcription initiation site, whereas a far larger proportion (approximately 25%) is potentially transcribable by elongating RNA polymerases. This observation raises the question of how the genome is packaged into chromatin to allow start sites to be recognized by the regulatory machinery at the same time as transcription initiation, but not elongation, is blocked in the 25% of intragenic DNA. We developed a chromatin scanning technique called ChAP, coupling the chromatin immunoprecipitation assay with arbitrarily primed PCR, which allows for the rapid and unbiased comparison of histone modification patterns within the eukaryotic nucleus. Methylated lysine 4 (K4) and acetylated K9/14 of histone H3 were both highly localized to the 5' regions of transcriptionally active human genes but were greatly decreased downstream of the start sites. Our results suggest that the large transcribed regions of human genes are maintained in a deacetylated conformation in regions read by elongating polymerase. Common models depicting widespread histone acetylation and K4 methylation throughout the transcribed unit do not therefore apply to the majority of human genes. PMID- 15123804 TI - The soluble ectodomain of herpes simplex virus gD contains a membrane-proximal pro-fusion domain and suffices to mediate virus entry. AB - Entry of herpes simplex virus (HSV) 1 into cells requires the interaction of HSV gD with herpesvirus entry mediator or nectin1 receptors, and fusion with cell membrane mediated by the fusion glycoproteins gB, gH, and gL. We report that the gD ectodomain in soluble form (amino acids 1-305) was sufficient to rescue the infectivity of a gD-null HSV mutant, indicating that gD does not need to be anchored to the virion envelope to mediate entry. Entry mediated by soluble gD required, in addition to the receptor-binding sites contained within residues 1 250, a discrete downstream portion (amino acids 261-305), located proximal to the transmembrane segment in full-length gD. We named it as profusion domain. The pro fusion domain was required for entry mediated by virion-bound gD, because its substitution with the corresponding region of CD8 failed to complement the infectivity of gD(-/+) HSV. Furthermore, a receptor-negative gD (gD(Delta6-259)) inhibited virus infectivity when coexpressed with wild-type gD; i.e., it acted as a dominant-negative gD mutant. The pro-fusion domain is proline-rich, which is characteristic of regions involved in protein-protein interactions. P291L-P292A substitutions diminished the gD capacity to complement gD(-/+) HSV infectivity. We propose that gD forms a tripartite complex with its receptor and, by way of the proline-rich pro-fusion domain, with the fusion glycoproteins, or with one of them. The tripartite complex would serve to recruit/activate the fusion glycoproteins and bring them from a fusion-inactive to a fusion-active state, such that they execute fusion of the virion envelope with cell membrane. PMID- 15123805 TI - Chromatin immunoprecipitation microarrays for identification of genes silenced by histone H3 lysine 9 methylation. AB - Switching from acetylation to methylation at histone H3 lysine 9 (K9) has recently been shown to contribute to euchromatin gene silencing. To identify genes silenced by K9 modifications, we probed a human CpG island microarray with DNA obtained by chromatin immunoprecipitation (ChIP) in a cancer cell line using an anti-H3-K9 methylated antibody or an anti-H3-K9 acetylated antibody. Of the 27 clones with the highest signal ratio of K9 methylation over acetylation (Me/Ac), 13 contained repetitive sequences. Among 14 nonrepetitive clones, we identified 11 genes (seven known and four previously undescribed), one EST, and two unknown fragments. Using ChIP-PCR, all 18 examined clones showed higher ratios of H3-K9 Me/Ac than the active gene control, P21, thus confirming the microarray data. In addition, we found a strong correlation between the K9 Me/Ac ratio and CpG island DNA methylation (R = 0.92, P < 0.01), and five of seven genes examined (megalin, thrombospondin-4, KR18, latrophilin-3, and phosphatidylinositol-3-OH kinase P101 subunit) showed lack of expression by RT-PCR and reactivation by DNA methylation and/or histone deacetylase inhibition, suggesting that these genes are true targets of silencing through histone modifications. All five genes also showed significant DNA methylation in a cell line panel and in primary colon cancers. Our data suggest that CpG island microarray coupled with ChIP can identify novel targets of gene silencing in cancer. This unbiased approach confirms the tight coupling between DNA methylation and histone modifications in cancer and could be used to probe gene silencing in nonneoplastic conditions as well. PMID- 15123806 TI - Paracrine regulation of mammalian oocyte maturation and male germ cell survival. AB - Mammalian oocytes are arrested at the prophase of meiosis before induction of maturation by the preovulatory luteinizing hormone (LH) surge. LH also promotes the survival of meiotic male germ cells in the testis. Because LH binds somatic cells, the mechanism underlying its regulation of germ cell function is unclear. We found that LH stimulates Leydig insulin-like 3 (INSL3) transcripts in ovarian theca and testicular Leydig cells. INSL3, in turn, binds a G protein-coupled receptor, LGR8 (leucine-rich repeat-containing G protein-coupled receptor 8), expressed in germ cells to activate the inhibitory G protein, thus leading to decreases in cAMP production. Treatment with INSL3 initiates meiotic progression of arrested oocytes in preovulatory follicles in vitro and in vivo and suppresses male germ cell apoptosis in vivo, thus demonstrating the importance of the INSL3 LGR8 paracrine system in mediating gonadotropin actions. PMID- 15123807 TI - A small-molecule approach to studying invasive mechanisms of Toxoplasma gondii. AB - Toxoplasma gondii is the most common protozoan parasite of humans. Infection with T. gondii can lead to life-threatening disease as a result of repeated cycles of host cell invasion, parasite replication, and host cell lysis. Relatively little is known about the invasive mechanisms of T. gondii and related parasites within the Phylum Apicomplexa (including Plasmodium spp., the causative agents of malaria), due to difficulties associated with studying genes essential to invasion in haploid obligate intracellular organisms. To circumvent this problem, we have developed a high-throughput microscope-based assay, which we have used to screen a collection of 12,160 structurally diverse small molecules for inhibitors of T. gondii invasion. A total of 24 noncytotoxic invasion inhibitors were identified. Secondary assays demonstrated that different inhibitors perturb different aspects of invasion, including gliding motility, secretion of host cell adhesins from apical organelles (the micronemes), and extension of a unique tubulin-based structure at the anterior of the parasite (the conoid). Unexpectedly, the screen also identified six small molecules that dramatically enhance invasion, gliding motility, and microneme secretion. The small molecules identified here reveal a previously unrecognized complexity in the control of parasite motility and microneme secretion, and they constitute a set of useful probes for dissecting the invasive mechanisms of T. gondii and related parasites. Small-molecule-based approaches provide a powerful means to address experimentally challenging problems in host-pathogen interaction, while simultaneously identifying new potential targets for drug development. PMID- 15123808 TI - Helicobacter pylori interacts with the human single-domain trefoil protein TFF1. AB - Why Helicobacter pylori colonizes only gastric tissue is unknown. It is found on gastric mucus-secreting cells and in the overlying gastric mucus but not deep in gastric glands. This localization mirrors the expression of trefoil factor 1, TFF1. We hypothesized that H. pylori interacting with TFF1 could explain the tropism of this bacteria for gastric tissue. Recombinant human TFF1 expressed in Escherichia coli was purified by affinity chromatography, ion-exchange chromatography, and gel filtration. Binding of H. pylori was assessed by using flow cytometry and the BIAcore system, which allows real-time monitoring of molecular interactions. In flow cytometry, H. pylori bound to the TFF1 dimer, but Campylobacter jejuni strains and the laboratory strain of E. coli, HB101, did not bind. When the BIAcore system was used, H. pylori bound strongly to TFF1-coated dextran chips compared with uncoated chips. Binding was inhibited by a TFF1 monoclonal antibody and by soluble TFF1. H. pylori bound to porcine gastric mucin only if it was pretreated with TFF1. In conclusion, H. pylori interacts avidly with the dimeric form of TFF1, and this interaction enables binding to gastric mucin, suggesting that TFF1 may act as a receptor for the organism in vivo. This interaction may underline the previously unexplained tropism of this organism for gastric tissue and its colocalization with the gastric mucin MUC5AC. PMID- 15123809 TI - Identification of core amino acids stabilizing rhodopsin. AB - Rhodopsin is the only G protein-coupled receptor (GPCR) whose 3D structure is known; therefore, it serves as a prototype for studies of the GPCR family of proteins. Rhodopsin dysfunction has been linked to misfolding, caused by chemical modifications that affect the naturally occurring disulfide bond between C110 and C187. Here, we identify the structural elements that stabilize rhodopsin by computational analysis of the rhodopsin structure and comparison with data from previous in vitro mutational studies. We simulate the thermal unfolding of rhodopsin by breaking the native-state hydrogen bonds sequentially in the order of their relative strength, using the recently developed Floppy Inclusion and Rigid Substructure Topography (FIRST) method [Jacobs, D. J., Rader, A. J., Kuhn, L. A. & Thorpe, M. F. (2001) Proteins 44, 150-165]. Residues most stable under thermal denaturation are part of a core, which is assumed to be important for the formation and stability of folded rhodopsin. This core includes the C110-C187 disulfide bond at the center of residues forming the interface between the transmembrane and the extracellular domains near the retinal binding pocket. Fast mode analysis of rhodopsin using the Gaussian network model also identifies the disulfide bond and the retinal ligand binding pocket to be the most rigid region in rhodopsin. Experiments confirm that 90% of the amino acids predicted by the FIRST method to be part of the core cause misfolding upon mutation. The observed high degree of conservation (78.9%) of this disulfide bond across all GPCR classes suggests that it is critical for the stability and function of GPCRs. PMID- 15123810 TI - The diploid genome sequence of Candida albicans. AB - We present the diploid genome sequence of the fungal pathogen Candida albicans. Because C. albicans has no known haploid or homozygous form, sequencing was performed as a whole-genome shotgun of the heterozygous diploid genome in strain SC5314, a clinical isolate that is the parent of strains widely used for molecular analysis. We developed computational methods to assemble a diploid genome sequence in good agreement with available physical mapping data. We provide a whole-genome description of heterozygosity in the organism. Comparative genomic analyses provide important clues about the evolution of the species and its mechanisms of pathogenesis. PMID- 15123811 TI - Imaging single membrane fusion events mediated by SNARE proteins. AB - Using total internal reflection fluorescence microscopy, we have developed an assay to monitor individual fusion events between proteoliposomes containing vesicle soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) and a supported planar bilayer containing cognate target SNAREs. Approach, docking, and fusion of individual vesicles to the target membrane were quantified by delivery and subsequent lateral spread of fluorescent phospholipids from the vesicle membrane into the target bilayer. Fusion probability was increased by raising divalent cations (Ca2+ and Mg2+). Fusion of individual vesicles initiated in <100 ms after the rise of Ca2+ and membrane mixing was complete in 300 ms. Removal of the N-terminal H(abc) domain of syntaxin 1A increased fusion probability >30-fold compared to the full-length protein, but even in the absence of the H(abc) domain, vesicle fusion was still enhanced in response to Ca2+ increase. Our observations establish that the SNARE core complex is sufficient to fuse two opposing membrane bilayers at a speed commensurate with most membrane fusion processes in cells. This real-time analysis of single vesicle fusion opens the door to mechanistic studies of how SNARE and accessory proteins regulate fusion processes in vivo. PMID- 15123812 TI - Incorporating chemical modification constraints into a dynamic programming algorithm for prediction of RNA secondary structure. AB - A dynamic programming algorithm for prediction of RNA secondary structure has been revised to accommodate folding constraints determined by chemical modification and to include free energy increments for coaxial stacking of helices when they are either adjacent or separated by a single mismatch. Furthermore, free energy parameters are revised to account for recent experimental results for terminal mismatches and hairpin, bulge, internal, and multibranch loops. To demonstrate the applicability of this method, in vivo modification was performed on 5S rRNA in both Escherichia coli and Candida albicans with 1-cyclohexyl-3-(2-morpholinoethyl) carbodiimide metho-p-toluene sulfonate, dimethyl sulfate, and kethoxal. The percentage of known base pairs in the predicted structure increased from 26.3% to 86.8% for the E. coli sequence by using modification constraints. For C. albicans, the accuracy remained 87.5% both with and without modification data. On average, for these sequences and a set of 14 sequences with known secondary structure and chemical modification data taken from the literature, accuracy improves from 67% to 76%. This enhancement primarily reflects improvement for three sequences that are predicted with <40% accuracy on the basis of energetics alone. For these sequences, inclusion of chemical modification constraints improves the average accuracy from 28% to 78%. For the 11 sequences with <6% pseudoknotted base pairs, structures predicted with constraints from chemical modification contain on average 84% of known canonical base pairs. PMID- 15123813 TI - CD81 is an entry coreceptor for hepatitis C virus. AB - Hepatitis C virus (HCV) envelope glycoproteins E1/E2 can pseudotype retroviral particles and efficiently mediate entry into target cells. Using this experimental system, we determined HCV tropism for different cell types. Only primary hepatocytes and one hepatoma cell line were susceptible to HCV pseudovirus entry, which could be inhibited by sera from HCV-infected individuals. Furthermore, expression of the putative HCV receptor CD81 on nonpermissive human hepatic but not murine cells enabled HCV pseudovirus entry. Importantly, inhibition of viral entry by an anti-CD81 mAb occurred at a step following HCV attachment to target cells. Our results indicate that CD81 functions as a post-attachment entry coreceptor and that other cellular factors act in concert with CD81 to mediate HCV binding and entry into hepatocytes. PMID- 15123814 TI - A random-periods model for expression of cell-cycle genes. AB - We propose a nonlinear regression model for quantitatively analyzing periodic gene expression in studies of experimentally synchronized cells. Our model accounts for the observed attenuation in cycle amplitude by a simple and biologically plausible mechanism. We represent the expression level for each gene as an average across a large number of cells. For a given cell-cycle gene, we model its expression in each cell in the culture as following the same sinusoidal function except that the period, which in any individual cell must be the same for all cell-cycle genes, varies randomly across cells. We model these random periods by using a lognormal distribution. The variability in period causes the measured amplitude of the cyclic expression trajectory to attenuate over time as cells fall increasingly out of synchrony. Gene-specific parameters include initial amplitude and phase angle. Applying the model to data from Whitfield et al. [Whitfield, M. L., Sherlock, G., Saldanha, A. J., Murray, J, I., Ball, C. A., et al. (2002) Mol. Biol. Cell 13, 1977-2000], we fit the trajectories of 18 well characterized phase-marker genes and find that the fit does not suffer when a common lognormal distribution is assumed for all 18 genes compared with a separate distribution for each. We then use the model to identify 337 periodically expressed transcripts, including the 18 phase-marker genes. The model permits estimation of and hypothesis testing about biologically meaningful parameters that characterize cycling genes. PMID- 15123815 TI - RNA surveillance down-regulates expression of nonfunctional kappa alleles and detects premature termination within the last kappa exon. AB - Random V(D)J junctions would generate nonfunctional and/or out-of-frame sequences in about two-thirds of cases and result in abundant transcripts encoding truncated proteins. Although allelic exclusion at the DNA recombination level ensures that a single allele is functional, the frequent biallelic rearrangements need additional mechanisms to down-regulate aberrant transcripts in those cells with both a functionally and a nonfunctionally rearranged allele. The process of nonsense-mediated decay targets aberrantly rearranged Ig heavy-chain transcripts, but the situation of light-chain mRNAs is more complex, because they do not meet the usual requirements for nonsense-mediated decay and most often lack a spliceable intron downstream of the premature termination. We studied immunoglobulin heavy-chain -/- pro-B cells in which light chain genes get rearranged and expressed in the absence of any selection for the assembly of a functional B cell receptor. Using this model, we show that the whole kappa locus is accessible in pro-B cells and allows the assembly of a broad spectrum of VkappaJkappa segments, most of which are out-of-frame. This model provides an evaluation of the in vivo efficiency of RNA surveillance toward aberrant kappa mRNAs produced in pro-B cells. Our data show that nonfunctional kappa transcripts are excluded from the mature mRNA pool not only by detecting termination in an upstream exon but also by detecting changes in the position of termination within the last exon. Similar mechanisms efficiently down-regulate nonfunctional kappa transcripts arising in normal mature B cells due to the biallelic transcription of rearranged kappa genes. PMID- 15123816 TI - The unfolding kinetics of ubiquitin captured with single-molecule force-clamp techniques. AB - We use single-molecule force spectroscopy to study the kinetics of unfolding of the small protein ubiquitin. Upon a step increase in the stretching force, a ubiquitin polyprotein extends in discrete steps of 20.3 +/- 0.9 nm marking each unfolding event. An average of the time course of these unfolding events was well described by a single exponential, which is a necessary condition for a memoryless Markovian process. Similar ensemble averages done at different forces showed that the unfolding rate was exponentially dependent on the stretching force. Stretching a ubiquitin polyprotein with a force that increased at a constant rate (force-ramp) directly measured the distribution of unfolding forces. This distribution was accurately reproduced by the simple kinetics of an all-or-none unfolding process. Our force-clamp experiments directly demonstrate that an ensemble average of ubiquitin unfolding events is well described by a two state Markovian process that obeys the Arrhenius equation. However, at the single molecule level, deviant behavior that is not well represented in the ensemble average is readily observed. Our experiments make an important addition to protein spectroscopy by demonstrating an unambiguous method of analysis of the kinetics of protein unfolding by a stretching force. PMID- 15123817 TI - p300 regulates p53-dependent apoptosis after DNA damage in colorectal cancer cells by modulation of PUMA/p21 levels. AB - Activation of the tumor suppressor p53 by DNA damage induces either cell cycle arrest or apoptosis, but what determines the choice between cytostasis and death is not clear. In this report, we show that the E1A-binding p300 nucleoprotein is a key determinant of p53-dependent cell fate in colorectal cancer cells: absence of p300 increases apoptosis in response to DNA damage. In addition, p300 deficient (p300(-)) cells fail to undergo G(1)/S arrest after UV irradiation. These abnormalities are associated with prolongation of p53 stability, reduced p53-acetylation, blunting of MDM2 activation, failure to transactivate p21, and a disproportionate increase in PUMA levels. When xenografted, p300(-) cells are more sensitive to chemotherapy with doxorubicin. These results show that p300 is a key regulator of the p53 response and suggest that p300 inhibition could be used to modulate chemotherapy. PMID- 15123818 TI - Microtubule composition: cryptography of dynamic polymers. PMID- 15123819 TI - TLR7: A new sensor of viral infection. PMID- 15123820 TI - Functional evidence for active site location of tetrameric thymidylate synthase X at the interphase of three monomers. AB - Little is known about the catalytic mechanism of the recently discovered ThyX family of flavin-dependent thymidylate synthases that are required for thymidylate (deoxythymidine 5'-monophosphate) synthesis in a large number of microbial species. Using a combination of site-directed mutagenesis and biochemical measurements, we have identified several residues of the Helicobacter pylori ThyX protein with crucial roles in ThyX catalysis. By providing functional evidence that the active site(s) of homotetrameric ThyX proteins is formed by three different subunits, our findings suggest that ThyX proteins have evolved through multimerization of inactive monomers. Moreover, because the active-site configurations of ThyX proteins, present in many human pathogenic bacteria, and of human thymidylate synthase ThyA are different, our results will aid in the identification of compounds specifically inhibiting microbial growth. PMID- 15123821 TI - Developmentally regulated IkappaB expression in intestinal epithelium and susceptibility to flagellin-induced inflammation. AB - Necrotizing enterocolitis is a devastating inflammatory condition of the intestine that occurs almost exclusively in premature newborns. Although its exact pathogenesis is unclear, we have postulated that it may result from a predisposition of the immature intestine to mount an unusually robust and damaging response to microbial infection. In support of this idea, we report that the IL-8 response of an immature human enterocyte cell line to bacterial infection was significantly higher than that of a mature enterocyte cell line. The response in both cell lines was flagellin-dependent. Corresponding to the difference in IL-8 production, the immature enterocytes expressed appreciably lower levels of specific IkappaB genes when compared with the mature enterocytes. Similar developmentally regulated differences in cytokine response and IkappaB expression were also seen in primary rat enterocytes, indicating that these observations were not peculiarities of the cell lines. Furthermore, when the level of IkappaBalpha expression was increased in the immature cell line by transfection, the flagellin-dependent IL-8 response was attenuated. Thus, we have demonstrated a previously undescribed developmental regulation of IkappaB expression in the intestine involved in modulating the IL-8 response to bacterial infection, which may contribute to the pathogenesis of age-specific inflammatory bowel diseases such as necrotizing enterocolitis. PMID- 15123822 TI - Turning on ribonucleotide reductase by light-initiated amino acid radical generation. AB - Ribonucleotide reductases (RNRs) catalyze the conversion of nucleotides to deoxynucleotides in all organisms, providing the monomeric precursors required for DNA replication and repair. The class I RNRs are composed of two subunits; the R1 subunit contains the active site for nucleotide reduction and allosteric effector binding sites, whereas the R2 subunit houses the essential diirontyrosyl (Y.) radical cofactor. A major unresolved issue is the mechanism by which the tyrosyl radical on R2 (Y122, Escherichia coli numbering) reversibly generates the transient thiyl radical (S.) on R1 that initiates nucleotide reduction. This intersubunit radical initiation is postulated to occur through a defined pathway involving conserved aromatic amino acids (R2: Y122, W48, Y356; R1: Y731, Y730) over a long distance of 35 A. A 20-mer peptide identical to the C-terminal tail of R2 (356-375) and containing Y356 is a competitive inhibitor with respect to R2, and it effectively blocks nucleotide reduction. We now report that a 21-mer peptide, in which a tryptophan has been incorporated at the N terminus of the 20th mer, can replace the R2 subunit and initiate nucleotide reduction by photoinitiated radical generation. The deoxynucleotide generated depends on the presence of allosteric effector and is pathway-dependent. Replacement of Y731 of R2 with phenylalanine prevents deoxynucleotide formation. These results provide direct evidence for the chemical competence of aromatic amino acid radicals and the importance of Y356 in R2 in the radical initiation process of the class I RNRs. PMID- 15123823 TI - Interactions of glutaredoxins, ribonucleotide reductase, and components of the DNA replication system of Escherichia coli. AB - A strain of Escherichia coli missing three members of the thioredoxin superfamily, thioredoxins 1 and 2 and glutaredoxin 1, is unable to grow, a phenotype presumed to be due to the inability of cells to reduce the essential enzyme ribonucleotide reductase. Two classes of mutations can restore growth to such a strain. First, we have isolated a collection of mutations in the gene for the protein glutaredoxin 3 that suppress the growth defect. Remarkably, all eight independent mutations alter the same amino acid, methionine-43, changing it to valine, isoleucine, or leucine. From the position of the amino acid changes and their effects, we propose that these alterations change the protein so that its properties are closer to those of glutaredoxin 1. The second means of suppressing the growth defects of the multiply mutant strain was by mutations in the DNA replication genes, dnaA and dnaN. These mutations substantially increase the expression of ribonucleotide reductase, most likely by altering the interaction of the regulatory protein DnaA with the ribonucleotide reductase promoter. Our results suggest that this increase in the concentration of ribonucleotide reductase in the cell allows more effective interaction with glutaredoxin 3, thus restoring an effective pool of deoxyribonucleotides. Our studies present direct evidence that ribonucleotide reductase is the only essential enzyme that requires the three reductive proteins missing in our strains. Our results also suggest an unexpected regulatory interaction between the DnaA and DnaN proteins. PMID- 15123824 TI - Latest folding game results: protein A barely frustrates computationalists. PMID- 15123825 TI - Malondialdehyde adducts in DNA arrest transcription by T7 RNA polymerase and mammalian RNA polymerase II. AB - Malondialdehyde, a genotoxic byproduct of lipid peroxidation, reacts with guanine in DNA to form pyrimido[1,2-alpha]purin-10(3H)one (M(1)dG), the first endogenous DNA lesion found to be a target of nucleotide excision repair enzymes. A subpathway of nucleotide excision repair, transcription-coupled repair, is thought to occur when RNA polymerase (RNAP) is arrested at damage in transcribed DNA strands and might function for efficient removal of M(1)dG in active genes. Results presented here show that M(1)dG and its stable, exocyclic analog 1,N(2) propanodeoxyguanine (PdG), arrest translocation of T7 RNAP and mammalian RNAPII when located in the transcribed strand of a DNA template. M(1)dG paired with thymine is exocyclic and poses a stronger block to transcription than the acyclic N(2)-(3-oxo-1-propenyl)-dG, formed upon cytosine-catalyzed opening of M(1)dG in duplex DNA. PdG is a complete block to RNAPII regardless of base pairing. The elongation factor TFIIS (SII) induces reversal and RNA transcript cleavage by RNAPII arrested at PdG. Thus, arrested RNAPII complexes may be stable at M(1)dG in cells and may resume transcription once the offending adduct is removed. The conclusion from this work is that malondialdehyde adducts in the transcribed strand of expressed genes are strong blocks to RNAPs and are targets for cellular transcription-coupled repair. If so, then M(1)dG, already known to be highly mutagenic in human cells, also may contribute to apoptosis in the developing tissues of individuals with Cockayne's syndrome, a hereditary disorder characterized by transcription-coupled repair deficiency. PMID- 15123826 TI - DNA end joining becomes less efficient and more error-prone during cellular senescence. AB - Accumulation of somatic mutations is thought to contribute to the aging process. Genomic instability has been shown to increase during aging, suggesting an aberrant function of DNA double-strand break (DSB) repair. Surprisingly, DSB repair has not been examined with respect to cellular senescence. Therefore, we have studied the ability of young, presenescent, and senescent normal human fibroblasts to repair DSBs in transfected DNA by using a fluorescent reporter substrate. We have found that the efficiency of end joining is reduced up to 4.5 fold in presenescent and senescent cells, relative to young cells. Sequence analysis of end junctions showed that the frequency of precise ligation was higher in young cells, whereas end joining in old cells was associated with extended deletions. These results indicate that end joining becomes inefficient and more error-prone during cellular senescence. Furthermore, the ability to use microhomologies for end joining was compromised in senescent cells, suggesting that young and senescent cells may use different end joining pathways. We hypothesize that inefficient and aberrant end joining is a likely mechanism underlying the age-related genomic instability and higher incidence of cancer in the elderly. PMID- 15123827 TI - Imprinting of the human L3MBTL gene, a polycomb family member located in a region of chromosome 20 deleted in human myeloid malignancies. AB - L3MBTL encodes a member of the Polycomb family of proteins, which, together with Trithorax group proteins, is responsible for the coordinated regulation of patterns of gene activity. Members of the Polycomb family also regulate self renewal of normal and malignant hematopoietic stem cells. L3MBTL lies in a region of chromosome 20, deletion of which is associated with myeloid malignancies and represents a good candidate for a 20q target gene. However, mutations of L3MBTL have not been identified in patients with 20q deletions or in cytogenetically normal patients. Here we demonstrate that monoallelic methylation of two CpG islands correlates with transcriptional silencing of L3MBTL, and that L3MBTL transcription occurs from the paternally derived allele in five individuals from two families. Expression of the paternally derived allele was observed in multiple hematopoietic cell types as well as in bone marrow derived mesenchymal cells. Deletions of 20q associated with myeloid malignancies resulted in loss of either the unmethylated or methylated allele. Our results demonstrate that L3MBTL represents a previously undescribed imprinted locus, a vertebrate Polycomb group gene shown to be regulated by this mechanism, and has implications for the pathogenesis of myeloid malignancies associated with 20q deletions. PMID- 15123828 TI - Application of evolutionary algorithm methods to polypeptide folding: comparison with experimental results for unsolvated Ac-(Ala-Gly-Gly)5-LysH+. AB - We present an evolutionary method for finding the low-energy conformations of polypeptides. The application, called FOLDAWAY,is based on a generic framework and uses several evolutionary operators as well as local optimization to navigate the complex energy landscape of polypeptides. It maintains two complementary representations of the structures and uses the CHARMM force field for evaluating the energies. The method is applied to unsolvated Met-enkephalin and Ac-(Ala-Gly Gly)(5)-Lys(+)H(+). Unsolvated Ac-(Ala-Gly-Gly)(5)-Lys(+)H(+) has been the object of recent experimental studies using ion mobility measurements. It has a flat energy landscape where helical and globular conformations have similar energies. FOLDAWAY locates several large groups of structures not found in previous molecular dynamics simulations for this peptide, including compact globular conformations, which are probably present in the experiments. However, the relative energies of the different conformations found by FOLDAWAY do not accurately match the relative energies expected from the experimental observations. PMID- 15123829 TI - CRE recombinase-inducible RNA interference mediated by lentiviral vectors. AB - Recently, several systems designed to trigger RNA interference by using small hairpin RNA driven by polymerase III promoters have been described. Here, we report a lentiviral-mediated small interfering RNA delivery system that can be induced by CRE recombinase. The system consists of a lentiviral vector carrying a mouse U6 promoter that is separated from a small hairpin RNA by a random DNA stuffer sequence flanked by modified loxP sites. The silencing cassette is not expressed until activated by addition of CRE recombinase delivered by a lentiviral vector. We have used this system to show specific down-regulation of GFP and two endogenous genes (the tumor suppressor p53 and the NF-kappaB transcription factor subunit p65) in vitro. Furthermore, down-regulation of both p53 and p65 resulted in the expected effect on downstream genes and cellular phenotype. We foresee multiple applications of this system both in vitro and in vivo to down-regulate specific targets in a tissue-specific and localized manner. PMID- 15123830 TI - New thioredoxin targets in the unicellular photosynthetic eukaryote Chlamydomonas reinhardtii. AB - Proteomics were used to identify the proteins from the eukaryotic unicellular green alga Chlamydomonas reinhardtii that can be reduced by thioredoxin. These proteins were retained specifically on a thioredoxin affinity column made of a monocysteinic thioredoxin mutant able to form mixed disulfides with its targets. Of a total of 55 identified targets, 29 had been found previously in higher plants or Synechocystis, but 26 were new targets. Biochemical tests were performed on three of them, showing a thioredoxin-dependent activation of isocitrate lyase and isopropylmalate dehydrogenase and a thioredoxin-dependent deactivation of catalase that is redox insensitive in Arabidopsis. In addition, we identified a Ran protein, a previously uncharacterized nuclear target in a photosynthetic organism. The metabolic and evolutionary implications of these findings are discussed. PMID- 15123831 TI - Single-molecule imaging analysis of Ras activation in living cells. AB - A single-molecule fluorescence resonance energy transfer (FRET) method has been developed to observe the activation of the small G protein Ras at the level of individual molecules. KB cells expressing H- or K-Ras fused with YFP (donor) were microinjected with the fluorescent GTP analogue BodipyTR-GTP (acceptor), and the epidermal growth factor-induced binding of BodipyTR-GTP to YFP-(H or K)-Ras was monitored by single-molecule FRET. On activation, Ras diffusion was greatly suppressed/immobilized, suggesting the formation of large, activated Ras signaling complexes. These complexes may work as platforms for transducing the Ras signal to effector molecules, further suggesting that Ras signal transduction requires more than simple collisions with effector molecules. GAP334-GFP recruited to the membrane was also stationary, suggesting its binding to the signaling complex. The single-molecules FRET method developed here provides a powerful technique to study the signal-transduction mechanisms of various G proteins. PMID- 15123832 TI - Hydration and mobility of HO-(aq). AB - The hydroxide anion plays an essential role in many chemical and biochemical reactions. But a molecular-scale description of its hydration state, and hence also its transport, in water is currently controversial. The statistical mechanical quasichemical theory of solutions suggests that HO.[H2O]3(-) is the predominant species in the aqueous phase under standard conditions. This result agrees with recent spectroscopic studies on hydroxide water clusters and with the available thermodynamic hydration free energies. In contrast, a recent ab initio molecular dynamics simulation has suggested that HO.[H2O]4(-) is the only dominant aqueous solution species. We apply adiabatic ab initio molecular dynamics simulations and find good agreement with both the quasichemical theoretical predictions and experimental results. The present results suggest a picture that is simpler, more traditional, but with additional subtlety. These coordination structures are labile but the tricoordinate species is the prominent case. This conclusion is unaltered with changes in the electronic density functional. No evidence is found for rate-determining activated interconversion of a HO.[H2O]4(-) trap structure to HO.[H2O]3(-) mediating hydroxide transport. The view of HO- diffusion as the hopping of a proton hole has substantial validity, the rate depending largely on the dynamic disorder of the water hydrogen-bond network. PMID- 15123833 TI - Genome-wide somatic hypermutation. AB - DNA mutagenesis is generally considered harmful. Yet activated B cells normally mutate the Ig loci. Because this somatic hypermutation is potentially dangerous, it has been hypothesized that mutations do not occur throughout the genome but instead are actively targeted to the Ig loci. Here we challenge this longstanding and widely accepted hypothesis. We demonstrate that hypermutation requires no Ig gene sequences. Instead, activation-induced cytidine deaminase and other trans acting hypermutation factors may function as general mutators. PMID- 15123834 TI - Response experiments for nonlinear systems with application to reaction kinetics and genetics. AB - A unified type of response experiment is suggested for complex systems made up of individual species (atoms, molecules, quasi-particles, biological organisms, etc.). We make the following assumptions: (i) some of the species may exist in two forms, labeled and unlabeled, respectively; (ii) the kinetic and transport properties of the labeled and unlabeled species are the same, respectively (neutrality assumption); (iii) the experiment preserves the total input and output fluxes; only the fractions of the labeled compounds in the input and output fluxes are varied. Under these circumstances a linear integral superposition law connects the fractions of labeled species in the input and output fluxes. This linear superposition law is valid for homogeneous and inhomogeneous systems and for systems with intrinsic (hidden) state variables; it arises from the neutrality condition and holds even though the underlying dynamics of the process may be highly nonlinear. Because this response law does not involve the linearization of the evolution equations it has great potential for the analysis of complex physical, chemical, and biological systems. We compare our approach with the linearization techniques used in biochemistry and genetics. We consider a simple reaction network involving replication, transformation, and disappearance steps and study the influence of experimental (measurement) and linearization errors on the evaluated values of rate coefficients. We show that the method involving the linearization of the kinetic equations leads to unpredictable results; because of the interference between measurement and linearization errors, either error compensation or error amplification occurs. Although our approach does not eliminate the effects of measurement errors, it leads to more consistent results. For a broad range of input fractions no error amplification or compensation occurs, and the error range for the rate coefficients is about the same as the error range of the measurements. PMID- 15123835 TI - Recruitment of a cytoplasmic response regulator to the cell pole is linked to its cell cycle-regulated proteolysis. AB - The response regulator CtrA, which silences the Caulobacter origin of replication and controls multiple cell cycle events, is specifically proteolyzed in cells preparing to initiate DNA replication. At the swarmer-to-stalked cell transition and in the stalked compartment of the predivisional cell, CtrA is localized to the cell pole just before its degradation. Analysis of the requirements for CtrA polar localization and CtrA proteolysis revealed that both processes require a motif within amino acids 1-56 of the CtrA receiver domain, and neither process requires CtrA phosphorylation. These results strongly suggest that CtrA polar localization is coupled to its cell cycle-regulated proteolysis. The polarly localized DivK response regulator promotes CtrA localization and proteolysis, but it does not directly recruit CtrA to the cell pole. Mutations in the divJ and pleC histidine kinases perturb the characteristic asymmetry of CtrA localization and proteolysis in the predivisional cell. We propose that polar recruitment of CtrA evolved to ensure that CtrA is degraded only in the stalked half of the predivisional cell, perhaps by localizing a proteolytic adaptor protein to the stalked pole. This is an example of controlled proteolysis of a cytoplasmic protein that is associated with its active recruitment to a specific subcellular address. PMID- 15123836 TI - Parallel somatic and synaptic processing in the induction of intermediate-term and long-term synaptic facilitation in Aplysia. AB - The induction of different phases of memory depends on the amount and patterning of training, raising the question of whether specific training patterns engage different cellular mechanisms and whether these mechanisms operate in series or in parallel. We examined these questions by using a cellular model of memory formation: facilitation of the tail sensory neuron-motor neuron synapses by serotonin (5-hydroxytryptamine, 5-HT) in the CNS of Aplysia. We studied facilitation in two temporal domains: intermediate-term facilitation (1.5-3 h) and long-term facilitation (LTF, >24 h). Both forms can be induced by using several different temporal and spatial patterns of 5-HT, including (i) repeated, temporally spaced pulses of 5-HT to both the sensory neuron soma and the sensory neuron-motor neuron synapse, and (ii) temporally asymmetric exposure of 5-HT to the soma and synapse under conditions in which neither exposure alone induces LTF. We first examined the protein and RNA synthesis requirements for LTF induced by these two patterns and found that asymmetric (but not repeated) 5-HT application induced LTF that required postsynaptic protein and RNA synthesis. We next focused on the patterning and protein synthesis requirements for intermediate-term facilitation. We found that intermediate-term facilitation (i) is induced locally at the synapse, (ii) requires multiple pulses of 5-HT, and (iii) requires synaptic protein synthesis. Our findings show that different temporal and spatial patterns of 5-HT induce specific temporal phases of long lasting facilitation in parallel by engaging different cellular and molecular mechanisms. PMID- 15123837 TI - Ribozyme knockdown functionally links a 1,25(OH)2D3 membrane binding protein (1,25D3-MARRS) and phosphate uptake in intestinal cells. AB - We used a ribozyme loss-of-function approach to demonstrate that the protein product of a cDNA encoding a multifunctional membrane-associated protein binds the seco-steroid 1,25(OH)(2)D(3) and transduces its stimulatory effects on phosphate uptake. These results are paralleled by studies in which the ability of the hormone to stimulate phosphate uptake in isolated chick intestinal epithelial cells is abolished by preincubation with Ab099 directed against the amino terminus of the protein. We now report the complete sequence of the cloned chicken cDNA for the 1,25D(3)-MARRS (membrane-associated, rapid-response steroid binding) protein and reveal it to be identical to the multifunctional protein ERp57. Functional studies showed that active ribozyme, but not a scrambled control, decreased specific membrane-associated 1,25(OH)(2)D(3) binding, but did not affect binding to the nuclear receptor for 1,25(OH)(2)D(3). Seco-steroid dependent stimulation of protein kinase C activity was diminished as 1,25D(3) MARRS protein levels were reduced in the presence of the ribozyme, as judged by Western blot analyses. Phosphate uptake in isolated cells is an index of intestinal phosphate transport that occurs during growth and maturation. Whereas cells and perfused duodena robustly responded to 1,25(OH)(2)D(3) in preparations from young birds, older animals no longer responded with stimulated phosphate uptake or transport. The age-related decline was accompanied by a decrease in 1,25D(3)-MARRS mRNA that was apparent up to 1 year of age. Together, these studies functionally link phosphate transport in the chick duodenum with the 1,25D(3)-MARRS protein and point to a previously uncharacterized role for this multifunctional protein class. PMID- 15123838 TI - Measuring the refolding of beta-sheets with different turn sequences on a nanosecond time scale. AB - Whether turns play an active or passive role in protein folding remains a controversial issue at this juncture. Here we use a photolabile cage strategy in combination with laser-flash photolysis and photoacoustic calorimetry to study the effects of different turns on the kinetics of beta-hairpin refolding on a nanosecond time scale. This strategy opens up a temporal window to allow the observation of early kinetic events in the protein refolding process at ambient temperature and pH without interference from any denaturants. Our results provide direct evidence demonstrating that even a one-residue difference in the turn region can change the refolding kinetics of a peptide. This observation suggests an active role for turn formation in directing protein folding. PMID- 15123839 TI - In vivo tracking of T cell development, ablation, and engraftment in transgenic zebrafish. AB - Transgenic zebrafish that express GFP under control of the T cell-specific tyrosine kinase (lck) promoter were used to analyze critical aspects of the immune system, including patterns of T cell development and T cell homing after transplant. GFP-labeled T cells could be ablated in larvae by either irradiation or dexamethasone added to the water, illustrating that T cells have evolutionarily conserved responses to chemical and radiation ablation. In transplant experiments, thymocytes from lck-GFP fish repopulated the thymus of irradiated wild-type fish only transiently, suggesting that the thymus contains only short-term thymic repopulating cells. By contrast, whole kidney marrow permanently reconstituted the T lymphoid compartment of irradiated wild-type fish, suggesting that long-term thymic repopulating cells reside in the kidney. PMID- 15123840 TI - The Six1 homeoprotein stimulates tumorigenesis by reactivation of cyclin A1. AB - Homeobox genes constitute a large family of transcription factors that are essential during normal development and are often dysregulated in cancer. However, the molecular mechanisms by which homeobox genes influence cancer remain largely unknown. Here we show that the tissue-restricted cyclin A1 is a transcriptional target of the Six1 homeoprotein. Both genes are expressed in the embryonic but not the terminally differentiated mammary gland, and Six1-knockout mice show a dramatic reduction of cyclin A1 in the embryonic mammary gland. In addition, both genes are reexpressed in breast cancers. Six1 overexpression increases cyclin A1 mRNA levels and activity, cell proliferation, and tumor volume, whereas Six1 down-regulation decreases cyclin A1 mRNA levels and proliferation. Overexpression of Six1 in wild-type mouse embryonic fibroblasts, but not in knockout variants lacking the cyclin A1 gene, induces cell proliferation. Furthermore, inhibition of cyclin A1 in Six1-overexpressing mammary carcinoma cells decreases proliferation. Together these results demonstrate that cyclin A1 is required for the proliferative effect of Six1. We conclude that Six1 overexpression reinstates an embryonic pathway of proliferation in breast cancer by up-regulating cyclin A1. PMID- 15123842 TI - What's "unstable" in unstable angina? AB - The role of emotional distress (e.g., anger, depression, and anxiety) in anginal chest discomfort (ACD) may have been underestimated. The authors review the empirical studies in this area, which are inconsistent with the standard theory on the ischemia-angina relationship; summarize the substantial evidence indicating a strong and consistent cross-sectional/prospective epidemiological association of emotional distress and ischemia/ACD; review the distress-targeted, interventional evidence confirming a causal relationship (i.e., reduced chest discomfort and health system utilization), thus confirming clinical utility of such interventions; and explore the possible mechanisms that might account for the relationship between emotional distress and chest discomfort. Substantial clinical benefit may be achieved by aggressively detecting and treating emotional distress in ACD patients. PMID- 15123841 TI - Requirement for a conserved Toll/interleukin-1 resistance domain protein in the Caenorhabditis elegans immune response. AB - The p38 mitogen-activated protein kinase pathway regulates innate immune responses in evolutionarily diverse species. We have previously shown that the Caenorhabditis elegans p38 mitogen-activated protein kinase, PMK-1, functions in an innate immune response pathway that mediates resistance to a variety of microbial pathogens. Here, we show that tir-1, a gene encoding a highly conserved Toll/IL-1 resistance (TIR) domain protein, is also required for C. elegans resistance to microbial pathogens. RNA interference inactivation of tir-1 resulted in enhanced susceptibility to killing by pathogens and correspondingly diminished PMK-1 phosphorylation. Unlike all known TIR-domain adapter proteins, overexpression of the human TIR-1 homologue, SARM, in mammalian cells was not sufficient to induce expression of NF-kappaB or IRF3-dependent reporter genes that are activated by Toll-like receptor signaling. These data reveal the involvement of a previously uncharacterized, evolutionarily conserved TIR domain protein in innate immunity that is functionally distinct from other known TIR domain signaling adapters. PMID- 15123843 TI - Relationships among alexithymia, adverse childhood experiences, sociodemographic variables, and actual mood disorder: a 2-year clinical follow-up study of patients with major depressive disorder. AB - This 2-year follow-up study examined relationships among alexithymia, adverse childhood experiences, sociodemographic variables, and actual mood disorder among patients with major depressive disorder (N=106). Alexithymia was assessed with the Toronto Alexithymia Scale (TAS-20), depression with the Beck Depression Inventory, and actual mood disorder with the Structured Clinical Interview for DSM-III-R. A questionnaire that assessed sociodemographic characteristics and adverse childhood experiences was also used. Long-lasting alexithymic features were associated with blue-collar work, harsh discipline, unhappiness of the childhood home, depression at 12 months, and major depressive disorder diagnosis at 24 months. Furthermore, the results showed that alexithymic features could also be situational reactions to depression. PMID- 15123844 TI - Amantadine for executive dysfunction syndrome in patients with dementia. AB - This article reports the results of an open uncontrolled chart review study of amantadine treatment for executive dysfunction syndrome in patients with dementia. All patients admitted to the neuropsychiatry or geriatric psychiatry inpatient units of Johns Hopkins Hospital in 2000 and 2001 who were treated empirically with amantadine for executive dysfunction syndrome were included in the review. Of the 30 patients whose cases were reviewed, 17 (57%) were at least "much improved," and most patients were discharged taking amantadine, suggesting that their physicians believed that they may have benefited from it. The medication was well tolerated in this frail group of patients. Most patients were taking one or more concurrent psychotropic medications, which may have contributed to the positive outcomes. Despite its limitations, this study offers preliminary data to support a controlled trial of amantadine in patients with executive dysfunction syndrome. PMID- 15123845 TI - Citalopram treatment of major depressive disorder in Hispanic HIV and AIDS patients: a prospective study. AB - Fourteen Hispanic and six non-Hispanic outpatients with HIV-spectrum illness and major depressive disorder were enrolled in a 6-week, open-label, flexible-dose study of citalopram (dose range=10-40 mg/day). The depressive symptoms of 50% of the 14 patients who completed the study responded to citalopram (mean dose=34 mg/day). The treatment response rate, effective citalopram dose, total number of reported adverse events, and attrition rate did not differ between the ethnic groups. Two patients discontinued because of adverse events (rash, nausea), and four patients discontinued because of noncompliance with the protocol. The findings suggest that citalopram is an effective and well-tolerated antidepressant for Hispanic and non-Hispanic HIV-infected patients. PMID- 15123846 TI - Safety and tolerability of extended-release venlafaxine in severe medical and surgical illness. AB - OBJECTIVE: This study attempted to determine if extended-release venlafaxine is safe for use in severely medically and surgically ill depressed patients. METHOD: The charts of 16 patients who were admitted to medical and surgical inpatient services and given extended-release venlafaxine were retrospectively evaluated for dose and duration of drug treatment, blood pressure changes, medication changes, and side effects. RESULTS: There was 50%-75% improvement in depressive symptoms, with a statistically insignificant mean elevation in blood pressure. CONCLUSIONS: Extended-release venlafaxine appears to be a safe and tolerable agent for the medical-surgical depressed inpatient. PMID- 15123847 TI - Developing a brief monitoring procedure for alcohol-dependent liver graft recipients. AB - To address the occurrence of deaths in later postoperative years among alcohol dependent liver graft recipients, the authors developed the Brief Active Focused Follow-Up protocol as an instrument for monitoring alcohol use following liver transplant. In this preliminary study, patient receptiveness to its use was tested and its ability to identify patient drinking was noted. Alcohol-dependent liver transplant recipients (N=24) and alcohol-dependent nontransplant patients (N=25) were asked to rate their receptiveness to the Brief Active Focused Follow Up in three areas. Subjects used a five-point scale for which 5 indicated the highest positive response. Liver transplant recipients responded positively to the Brief Active Focused Follow-Up, with mean responses of 4.92, 4.08, and 4.63 with regard to clarity, usefulness, and ease of completion, respectively. Nontransplant subjects responded similarly, with mean responses of 4.88, 4.12, and 4.52. The two groups were not significantly different in their receptiveness to the Brief Active Focused Follow-Up interview. The Brief Active Focused Follow Up identified alcohol use within the last 30 days: 8% (N=2 of 24) in the transplant group, and 56% (N=14 of 25) in the nontransplant group. These results suggest that 1) this manualized, brief monitoring technique is well received by alcohol-dependent liver transplant patients, and 2) the Brief Active Focused Follow-Up's "user friendliness" makes it a potentially appropriate instrument for long-term monitoring of alcohol use among alcohol-dependent liver graft recipients. PMID- 15123848 TI - Effect of medical comorbidity on response to fluoxetine augmentation or dose increase in outpatients with treatment-resistant depression. AB - This study assessed the effect of general medical comorbidity on response to next step antidepressant treatments among subjects with major depressive disorder whose depression failed to respond to an 8-week open trial of 20 mg/day of fluoxetine. Of the 386 outpatients in the open trial, 101 who remained depressed were randomly assigned to double-blind treatment with either an increased dose of fluoxetine or lithium or desipramine augmentation for 4 weeks. The Cumulative Illness Rating Scale (CIRS) was used to assess baseline general medical comorbidity, and the Hamilton Depression Rating Scale was used to assess depressive symptoms. Logistic regression analysis showed that CIRS score was not associated with likelihood of remission or premature study discontinuation. Medical comorbidity thus does not appear to be associated with significantly poorer outcome among patients whose major depressive disorder failed initially to respond to an initial trial of 20 mg/day of fluoxetine. PMID- 15123849 TI - Cognitive function in patients with chronic granulomatous disease: a preliminary report. AB - Chronic granulomatous disease is an inherited immunodeficiency in which phagocytes fail to generate superoxide and its metabolites, resulting in severe recurrent infections and frequent hospitalizations. Chronic illness and frequent hospitalizations can affect growth and development as well as social and educational opportunities. Since no data have been reported on cognitive functioning in patients with this illness, the authors sought to examine cognitive function in a group of patients with chronic granulomatous disease. A retrospective chart review of 26 patients seen and followed at the National Institutes of Health who had received cognitive testing at the request of parent or staff was performed. Demographic information including medical, psychiatric, and developmental histories was gathered. Six patients (23%) were found to have an IQ of 70 or below, indicative of cognitive deficits, and all of those patients had defects in the membrane-linked cytochrome b558. The prevalence of cognitive deficits in this selected population of chronic granulomatous disease patients was high. The determination of the true distribution of cognitive functioning in the general chronic granulomatous disease population is important, since cognitive deficits have implications for educational planning and potential therapies such as transplantation and gene therapy in children. PMID- 15123850 TI - How do delirium and dementia increase length of stay of elderly general medical inpatients? AB - The purpose of this study was to identify the proximate causes through which dementia and delirium extend length of stay (LOS) in elderly general hospital patients. Among 93 patients age >/=65 years admitted to a tertiary-care teaching hospital through the emergency department, admission ratings of cognitive impairment, delirium, and dementia predicted the emergence of mental and behavioral manifestations of delirium and dementia in the hospital and greater LOS. Mental and behavioral manifestations also predicted greater LOS. On average, mental manifestations appeared first and were followed by behavioral manifestations, and the appearance of both types of manifestations occurred before the mean LOS. The results suggest that elderly patients with dementia and/or delirium who become symptomatic after admission to a general hospital first show mental signs and symptoms, then show behavioral disturbances, which appear to be the proximate causes of greater LOS. PMID- 15123851 TI - Comorbid factitious and conversion disorders. PMID- 15123852 TI - Forgotten frontal lobe syndrome or "Executive Dysfunction Syndrome". PMID- 15123853 TI - False positives on the clinical institute withdrawal assessment for alcohol revised: is this scale appropriate for use in the medically ill? PMID- 15123854 TI - Antiretrovirals, part 1: overview, history, and focus on protease inhibitors. AB - This column is the first in a series on HIV/AIDS antiretroviral drugs. This first review summarizes the history of HIV/AIDS and the development of highly active antiretroviral therapy (HAART) and highlights why it is important for non-HIV specialists to know about these drugs. There are four broad classes of HIV medications used in varying combinations in HAART: the protease inhibitors, nucleoside analogue reverse transcriptase inhibitors, the non-nucleoside reverse transcriptase inhibitors, and cell membrane fusion inhibitors. This paper reviews the mechanism of action, side effects, toxicities, and drug interactions of the protease inhibitors. PMID- 15123855 TI - Citalopram and quality of life in lung transplant recipients. PMID- 15123857 TI - Mental health care in primary care settings. PMID- 15123859 TI - Child abuse. PMID- 15123860 TI - For a safer outcome with newborn jaundice. PMID- 15123861 TI - Efficacy of oral phenobarbitone in term "at risk" neonates in decreasing neonatal hyperbilirubinemia: a randomized double-blinded, placebo controlled trial. AB - OBJECTIVE: To evaluate the efficacy of oral phenobarbitone in "at risk " term neonates (with high cord bilirubin) in decreasing hyperbilirubinemia. DESIGN: Double blind, placebo-control, randomized trial. SETTING: Tertiary level neonatal unit. OUTCOME: Primary-hyperbilirubinemia defined as total serum bilirubin (TSB) greater than 13 mg/dL. Secondary-TSB at 72 +/- 12 hr, need for phototherapy or exchange transfusion and side effects of phenobarbitone therapy. METHODS: All consecutively born term healthy neonates with cord bilirubin > or = 2.5 mg/dL were randomly assigned to receive either phenobarbitone (n = 37) or placebo (n = 38) after obtaining informed consent. Phenobarbitone was administered orally (5 mg/kg/day) for 3 days starting within 12 hours of birth. The neonates were followed up till seven days of life. TSB was estimated in neonates who developed jaundice with clinically assessed level of 8-10 mg/dL and at 72 +/-12 hours of age in 55 neonates. RESULTS: The baseline characteristics were similar in two groups. There was no significant reduction in incidence of hyperbilirubinemia in phenobarbitone group compared to in placebo group (6/37 (16.2%) versus 13/38 (34.3%); RR 0.47, 95% confidence interval: 0.20-1.11; risk difference: -18.1%, 95% confidence interval: -39.5 to 3.3%). However TSB at 72 +/-12 hours in phenobarbitone group (mean +/- S.D: 10.0 +/- 3.7 mg/dL) was significantly lesser than in placebo group (mean +/- S.D: 12.3 +/- 3.3 mg/dL) (difference of means: 2.3 mg/dL, 95% confidence interval: -3.9 to -0.7 mg/dl, P = 0.018). No significant difference with respect to need for treatment was observed in two groups. No significant adverse effects of phenobarbitone were noted. CONCLUSIONS: Prophylactic phenobarbitone is not helpful in reducing the incidence of hyper bilirubinemia in "at risk" term neonates. PMID- 15123862 TI - Consensus Statement of IAP National Task Force: status report on management of acute diarrhea. PMID- 15123863 TI - Monitoring adverse reaction to steroid therapy in children. AB - Patients on corticosteroid therapy, specially for a long period are likely to develop many adverse effects related to the therapy. A physician should be conversant with these to ensure early detection, management and prevention, where possible. Thus, all patients on a long-term corticosteroid therapy should have a baseline and 3 monthly assessments for weight, height, blood pressure and other clinical features of Cushing's syndrome. A 2 hours postprandial blood sugar and serum electrolyte estimation should also be included. Ophthalmic evaluation for glaucoma and cataract should be carried out at 6 monthly intervals and densitometry annually for early detection of osteopenia. In addition, a high index of suspicion should be maintained for timely detection of infections, avascular bone necrosis, myopathy and pseudotumor cerebri. PMID- 15123864 TI - Aztreonam. AB - Aztreonam belongs to the monobactam group of naturally occurring antibiotic compounds characterized by a monocycling ring structure. Aztreonam is the first monobactam that has been approved for use in pediatric medicine by US FDA in the year 1998. PMID- 15123865 TI - Effect of partial exchange transfusion in asymptomatic polycythemic LBW babies. AB - This randomized controlled trial was conducted to determine the effect of partial exchange transfusion in polycythemic babies. Forty five asymptomatic polycythemic babies with birth weight < or = 2000 g were included and randomly assigned to undergo either partial exchange transfusion using isotonic saline within 4 hours of screening or routine medical management. Outcome measures were neonatal morbidity (especially hypoglycemia and neurological alterations) and mortality; developmental delays using DDST-II, neurological deficits, tone and DTR abnormalities over 18 months follow up period. The overall neonatal morbidity in this study was low and comparable in the two groups. Some of the polycythemic babies in the non-exchanged group found initially at 3 months age with "suspected development" grew out of their developmental delay at 18 months of age or later while those who underwent exchange transfusion and with retarded development at 3 months of age remained so even at 18 months of age. PMID- 15123866 TI - Pattern of pediatric dermatoses in a referral center in South India. AB - This study was undertaken to determine the pattern of dermatoses in children in south India. All children <14 years presenting to us between May 2001 and June 2002 were recruited. A total of 2100 children (males -995; females- 1105) with 2144 dermatoses were recorded. Infections and infestations were the most common dermatoses (54.5%) followed by dermatitis and eczema (8.6%), pigmentary disorders (5.7%), insect bite reaction (5.27%), hair and nail disorders (5.2%), miliaria (4.1%), nutritional deficiency disorders (2.8%), urticaria (2.5%), genetic disorders (2.1%), psoriasis (1.4%), collagen vascular disorders (0.5%), hemangiomas (0.5%), drug eruptions (0.3%), pityriasis rosea (0.2%) and others (5.8%). Pyodermas were the most common dermatoses (47.13%) followed by scabies (30.6%) amongst infections and infestations. Atopic dermatitis was noticed only in 3 patients. Insect bite reactions (papular urticaria) (5.27%) and miliaria (4.1%) were attributed to the tropical weather conditions in this coastal area. Genetic disorders including ichthyosis and palmoplantar keratoderma contributed to 2.1% of cases and could be due to the high incidence of consanguinous marriages in this society. PMID- 15123867 TI - Cost of syrup versus capsule form of vitamin A supplementation. AB - The study was done to estimate the cost of each dose of vitamin A (2,00,000 Units) to the health system when delivered as a capsule, applicap or as syrup form. The cost of distribution of vitamin A supplements was estimated for the manufacturers, district and delivery level. The lowest cost per dose was for capsules in plastic jar (Rs. 0.99) and the highest was for the syrup in glass bottle (Rs. 1.29), the option currently being practiced. The distribution costs were least for the capsule, which compensates for its higher production cost. The cost of syrup was also more due to high degree of wastage compared to capsules. While cost is an important issue, other operational factors need also to be considered. PMID- 15123868 TI - An outbreak of echovirus meningitis in children. AB - An outbreak of aseptic meningitis in children as evidenced by increase in the number of admissions in a tertiary care hospital is described. Clinical data and stool samples were collected from 25 hospitalized infants and young children. The stool samples were subjected to virological investigations. Fever and vomiting were the commonest symptoms. Cerebrospinal fluid (CSF) showed lymphocytic pleocytosis in majority of cases. Of the 25 stool samples, 14 showed an enterovirus specific cytopathogenic effect (CPE) in rhabdomyosarcoma (RD) cell line. All the 14 samples were positive for enterovirus RNA by reverse transcription-polymerase chain reaction (RT-PCR). Partial sequencing of the Virion protein 1 (VPI) region of the enterovirus genome carried out on the first 7 isolates revealed 5 isolates to be echovirus serotype 4 and one each to be echovirus serotypes 3 and 30. All children showed a rapid recovery and were discharged within 3 days of admission. PMID- 15123869 TI - Bronchial transection: delayed diagnosis and successful repair. AB - Traumatic bronchial transection is usually recognized and repaired immediately after injury. Bronchial transection has a variety of clinical presentations due to air leak into the pleural cavity and it is very rare to have total absence of air leak from the transected bronchus at presentation. We present one such case of main right bronchus injury with total absence of initial clinical signs and symptoms, leading to a delay in the diagnosis. However, the surgical repair eight months after injury showed excellent recovery of the chronically collapsed lung. PMID- 15123870 TI - Toxic epidermal necrolysis treated with intravenous immunoglobulin and granulocyte colony-stimulating factor. AB - We report a case of toxic epidermal necrolysis who was successfully treated with intravenous immunoglobin and granulocyte colony-stimulating factor. He had poor prognostic factors such as extensive epidermal loss, neutropenia, acute respiratory distress syndrome and candida sepsis, but nonetheless made a complete recovery. PMID- 15123871 TI - Acute renal tubular dysfunction in association with Salmonella enteritidis. AB - A thirteen-year-old boy presented with acute renal tubular dysfunction after an infection with salmonella enteritidis. The child recovered following treatment with ciprofloxacin for a week. PMID- 15123872 TI - Dengue shock syndrome in newborn: a case series. AB - Four cases of dengue shock syndrome were seen during an epidemic of dengue fever. Three cases recovered following appropriate management. PMID- 15123873 TI - Hypomelanosis of Ito. PMID- 15123874 TI - Fibrodysplasia ossificans progressiva. PMID- 15123875 TI - Travails of demystifying a mysterious disease in India. PMID- 15123876 TI - Blood lead levels among children aged 0-15 years in Hangzhou, China. PMID- 15123877 TI - Chronic renal failure with retinal detachment. PMID- 15123880 TI - Will Biotechnology Overcrowd the Planet? PMID- 15123879 TI - Familial infantile globoid cell leukodystrophy (Krabbes disease). PMID- 15123881 TI - National Cancer Institute and Imaging-Intersecting Scientific Opportunity with Clinical Need. PMID- 15123882 TI - Automatic Discrimination of Abnormal Subjects Using the Visual Evoked Potential Spectral Components. AB - Study of visual evoked potential (VEP) is one of the utilized methods in clinical diagnosis of ophthalmology and neurological disorders. The automatic detection of VEP spectral components is an important tool in the diagnosis of mental activity. This paper presents a novel computational approach using feedforward neural network to identify abnormal subjects from changes in spectral components. The output vector from the feedforward neural network is based on the VEP spectral components. The software was developed to identify mental state from the VEP spectral components using Matlab software package. Using this approach, it is possible to perform real-time abnormality identification accurately on personal computers. PMID- 15123883 TI - Patch Clamp Study of Serotonin-Gated Currents via 5-HT Type 3 Receptors by Using a Novel Approach SHAM for Receptor Channel Scanning. AB - We studied 5-hydroxy tryptamine type 3 (5-HT(3)) receptors transfected in tsA-201 cell line to examine serotonin-induced whole cell currents. Using the site directed mutagenesis technique, we individually mutated each residue in the membrane-spanning M2 segment to histidine. A high proportion of tsA-201 cells cotransfected with the cDNAs of 5-HT(3)R and CD8 produced large amplitude responses (0.5?7.0 nA) to serotonin. The dose-response curve of wild-type (WT) receptor ranging from 0.5 to 500 $?mu$ mole increases its K(d) values, and I_?mathrm{max} of 5-HT(3)R falls at low external pH as if protonation of an acid group is enough to block the channel. Lysine at position 281, a basic residue, is more susceptible to acidification-induced blockade of the 5-HT(3)R channel. Dose response curves of K281S (replacing lysine at the 281 position with serine) at different pH are not significantly modulated, and histidine substitutions at the three consecutive positions 293, 294, and 296 eliminate the pH block of the channel. PMID- 15123884 TI - Genetic Cross-Talk During Head Development in Drosophila. AB - The dorsal head vertex of Drosophila is specified mainly by the orthodenticle (otd) gene. The expression and the function of otd are regulated by the concerted action of many genes including hedgehog (hh) and notch (N). These genes are components of a meshwork of signaling transduction pathways that interact to form the dorsal head capsule of the fruit fly. Loss-of-function Hh mutants lack ocelli; however, loss-of-function N mutants lack a different domain of the dorsal head vertex. This report provides new evidence that the Hh and N pathways are two epistatic signaling cascades that act genetically upstream of the dorsal head capsule specification gene. PMID- 15123886 TI - Three-Dimensional Culture of Hybridoma Cells Secreting Anti-Human Chorionic Gonadotropin by a New Rolling Culture System. AB - Cell growth rate and production of monoclonal antibody (MAb) of hybridoma cells producing anti-human chorionic gonadotropin (hCG) MAb have been used as investigation criteria in double-mouthed rolling bottle (DMRB). Compared with T flask cell culture, both of the cell number and MAb production increased by approximately 42.5% when the medium was supplemented with 5% fetal calf serum (FCS) and DMRB rotated at 2 turns per minute. Yield of MAb was experimentally related to the number of viable cells. Interestingly, MAb yield was four times as high as that cultured in T-flask in the first 24 hours, and about 75% yield of total MAb was secreted by 48 hours during the culture. It appears that the promoted cell growth and MAb yield are resulted from the three-dimensional growth of hybridoma cells under a suitably revolving condition. PMID- 15123885 TI - Adhesion to Vitronectin and Collagen I Promotes Osteogenic Differentiation of Human Mesenchymal Stem Cells. AB - The mechanisms controlling human mesenchymal stem cells (hMSC) differentiation are not entirely understood. We hypothesized that the contact with extracellular matrix (ECM) proteins normally found in bone marrow would promote osteogenic differentiation of hMSC in vitro. To test this hypothesis, we cultured hMSC on purified ECM proteins in the presence or absence of soluble osteogenic supplements, and assayed for the presence of well-established differentiation markers (production of mineralized matrix, osteopontin, osteocalcin, collagen I, and alkaline phosphatase expression) over a 16-day time course. We found that hMSC adhere to ECM proteins with varying affinity (fibronectin > collagen I >/= collagen IV >/= vitronectin > laminin-1) and through distinct integrin receptors. Importantly, the greatest osteogenic differentiation occurred in cells plated on vitronectin and collagen I and almost no differentiation took place on fibronectin or uncoated plates. We conclude that the contact with vitronectin and collagen I promotes the osteogenic differentiation of hMSC, and that ECM contact alone may be sufficient to induce differentiation in these cells. PMID- 15123887 TI - Evidence for a Novel, Caspase-8-Independent, Fas Death Domain-Mediated Apoptotic Pathway. AB - Certain caspase-8 null cell lines demonstrate resistance to Fas-induced apoptosis, indicating that the Fas/FasL apoptotic pathway may be caspase-8 dependent. Some reports, however, have shown that Fas induces cell death independent of caspase-8. Here we provide evidence for an alternative, caspase-8 independent, Fas death domain-mediated apoptotic pathway. Murine 12B1-D1 cells express procaspase-3, -8, and -9, which were activated upon the dimerization of Fas death domain. Bid was cleaved and mitochondrial transmembrane potential was disrupted in this apoptotic process. All apoptotic events were completely blocked by the broad-spectrum caspase inhibitor Z-VAD-FMK, but not by other peptide caspase inhibitors. Cyclosporin A (CsA), which inhibits mitochondrial transition pore permeability, blocked neither pore permeability disruption nor caspase activation. However, CsA plus caspase-8 inhibitor blocked all apoptotic events of 12B1-D1 induced by Fas death domain dimerization. Our data therefore suggest that there is a novel, caspase-8-independent, Z-VAD-FMK-inhibitable, apoptotic pathway in 12B1-D1 cells that targets mitochondria directly. PMID- 15123888 TI - Definition of Soybean Genomic Regions That Control Seed Phytoestrogen Amounts. AB - Soybean seeds contain large amounts of isoflavones or phytoestrogens such as genistein, daidzein, and glycitein that display biological effects when ingested by humans and animals. In seeds, the total amount, and amount of each type, of isoflavone varies by 5 fold between cultivars and locations. Isoflavone content and quality are one key to the biological effects of soy foods, dietary supplements, and nutraceuticals. Previously we had identified 6 loci (QTL) controlling isoflavone content using 150 DNA markers. This study aimed to identify and delimit loci underlying heritable variation in isoflavone content with additional DNA markers. We used a recombinant inbred line (RIL) population ( $n=100$ ) derived from the cross of ?Essex? by ?Forrest,? two cultivars that contrast for isoflavone content. Seed isoflavone content of each RIL was determined by HPLC and compared against 240 polymorphic microsatellite markers by one-way analysis of variance. Two QTL that underlie seed isoflavone content were newly discovered. The additional markers confirmed and refined the positions of the six QTL already reported. The first new region anchored by the marker BARC Satt063 was significantly associated with genistein ( $P=0.009$, $Rcirc;2=29.5?%$ ) and daidzein ( $P=0.007$, $Rcirc;2=17.0?%$ ). The region is located on linkage group B2 and derived the beneficial allele from Essex. The second new region defined by the marker BARC-Satt129 was significantly associated with total glycitein ( $P=0.0005$, $Rcirc;2=32.0?%$ ). The region is located on linkage group D1a+Q and also derived the beneficial allele from Essex. Jointly the eight loci can explain the heritable variation in isoflavone content. The loci may be used to stabilize seed isoflavone content by selection and to isolate the underlying genes. PMID- 15123889 TI - Microencapsulated Genetically Engineered Lactobacillus plantarum 80 (pCBH1) for Bile Acid Deconjugation and Its Implication in Lowering Cholesterol. AB - Cholesterol is known to be a major risk factor for coronary heart disease (CHD). Current treatments for elevated blood cholesterol include dietary management, regular exercise, and drug therapy with fibrates, bile acid sequestrants, and statins. Such therapies, however, are often suboptimal and carry a risk for serious side effects. This study shows that microencapsulated Lactobacillus plantarum 80 (pCBH1) cells can efficiently break down and remove bile acids, and establishes a basis for their use in lowering blood serum cholesterol. Results show that microencapsulated LP80 (pCBH1) is able to effectively break down the conjugated bile acids glycodeoxycholic acid (GDCA) and taurodeoxycholic acid (TDCA) with bile salt hydrolase (BSH) activities of 0.19 and 0.08 $?mu$ mol DCA/mg CDW/h respectively. This article also summarizes the physiological interrelationship between bile acids and cholesterol and predicts the oral doses of microencapsulated Lactobacillus plantarum 80 (pCBH1) cells required for lowering cholesterol. PMID- 15123897 TI - Poisoning prevention education during emergency department visits for childhood poisoning. AB - OBJECTIVE: Childhood poisoning represents a major public health problem that can be prevented through educational efforts. Commonly, young children experiencing a first poisoning episode will have a second occurrence. The aim of this study is to assess whether caretakers of children evaluated in an emergency department for acute poisoning exposure recall receiving poisoning prevention education as part of the medical encounter. METHODS: Consecutive poisoning exposures reported to a regional Poison Control Center concerning children under the age of 6 years were identified. Inclusion criteria were an emergency department referral and subsequent discharge. Within 10 days of emergency department visit, a single trained interviewer administered by telephone a scripted questionnaire concerning the provision of poisoning prevention education. Poison control center logs were reviewed for demographic and treatment data. RESULTS: Of 102 eligible subjects, 77 families were contacted and 75 participated. The median age of poison exposure was 2 years; 45% were male. Seventy-three percent (95% CI: 61% to 83%) of caregivers reported receiving no poisoning prevention education. Among the minority receiving some information, only 25% received verbal instruction. Caregivers of children treated in urban academic centers were more likely to recall prevention education than those treated at suburban and rural hospitals (RR = 5.4; 1.8 to 16.2). Caregivers of children treated at specialized children's hospitals were more likely recall prevention education than general hospitals (RR = 5.3; 1.6 to 17.7). CONCLUSION: Emergency department personnel are missing a potentially important opportunity to provide poisoning prevention education to families of children at high risk for repeat occurrence. PMID- 15123898 TI - Acetaminophen dose accuracy and pediatric emergency care. AB - OBJECTIVE: To describe the demographic and clinical characteristics of children who receive acetaminophen per emergency department standing orders and identify factors associated with supratherapeutic doses (>or=16 mg/kg). METHODS: A cross sectional retrospective study of consecutive children who received acetaminophen at an urban children's hospital emergency department. RESULTS: Over a 1-week period, 156 (24%) of 661 emergency department children were treated with acetaminophen. Of the children receiving acetaminophen, 64 (41%) were less than 2 years and 92 (59%) were males. The indication for acetaminophen was fever in 140 (90%) children, and 70 (50%) had temperatures >or=39degreesC. The route of administration was oral [133 (85%)] and rectal [23 (15%)]. There were 122 (78%) correct doses (standing order dose range of 10 to 15 mg/kg), 15 (10%) doses of <10 mg/kg, and 19 (12%, 95% CI, 8% to 18%) doses of >or=16 mg/kg. In comparison, children who received acetaminophen at normal or low doses (<16 mg/kg) versus supratherapeutic doses (>or=16 mg/kg) showed a significant difference in route. The rate of rectal route was significantly greater in the supratherapeutic doses (95% CI for changes in proportion, 14% to 48%). CONCLUSIONS: Review of acetaminophen dose accuracy, particularly the rectal route, is recommended for emergency departments with standing orders for acetaminophen. PMID- 15123899 TI - Office preparedness for pediatric emergencies: baseline preparedness and the impact of guideline distribution. AB - OBJECTIVE: To determine the baseline level of preparedness of Wisconsin primary care physician offices for pediatric emergencies and to assess the impact of mail distribution of guidelines on this level of preparedness. METHOD: Preintervention surveys were mailed to all Wisconsin physicians requesting information about pediatric equipment, medications, training, and policies. Guidelines were distributed by mail to all physicians who returned completed surveys. An identical postintervention survey was mailed to these physicians, and an analysis of the paired surveys for each respondent was performed. RESULTS: Baseline preparedness of 1051 Wisconsin physician offices ranged from 37% with intraosseous needles to 96% with albuterol solution for inhalation. Physician certification in pediatric advanced life support was required in 26% of offices. A total of 568 paired preintervention and postintervention surveys were analyzed. Improvements were identified for the availability of equipment and medications, transport policies, and reference guides. CONCLUSIONS: Mail distribution of guidelines was minimally effective in improving the preparedness of Wisconsin offices for pediatric emergencies. PMID- 15123900 TI - Utility of an immunization registry in a pediatric emergency department. AB - OBJECTIVES: Determine prevalence of participation and underimmunization rate in a regional immunization registry (IR) among patients presenting to a university pediatric emergency department (PED). Rate of agreement between parental report and documented immunization status was also measured. METHODS: A convenience sample of parents of patients younger than 11 years registered in the PED were approached with a short questionnaire. When informed consent was obtained, the Central New York (CNY) IR was accessed via computer to see if the child was in the registry and to ascertain if their immunizations were up-to-date (UTD). Rate of agreement between parental report and immunization status documented in the IR was calculated. RESULTS: 698 (97%) of 720 patients consented to participate. Of these, 235 (34%, 95% CI, 30-37) were enrolled in the IR. Eighty-five (36%, 95% CI, 30-42) enrolled patients were under age 2. Sixty-seven (29%, 95% CI, 23-34) were from private group practices, 146 (62%, 95% CI, 56-68) were from university/community health center clinics and the source of primary care for 22 patients (9%) was unknown. Only 67 (29%, 95% CI, 23-34) parents of children in the IR were aware that they were enrolled. Of IR patients, 225 (96%, 95% CI, 93 98) stated they were UTD, while only 143 (61%, 95% CI, 55-67) were documented to be so. CONCLUSIONS: A significant number of patients seen in the PED were in the CNY IR. More than one-half of the parents of enrolled children did not recall that they had previously registered their child. Only 61% of patients were UTD, whereas parents reported that almost all were. In the PED, use of an IR would create an opportunity for intervention in a large number of patients who were not UTD. PMID- 15123901 TI - Into hot water head first: distribution of intentional and unintentional immersion burns. AB - INTRODUCTION: Experience with several, previously unreported, intentional face first immersion burns led us to evaluate the distribution of inflicted and unintentional immersion scald burns in a hospital series. SETTING: (1) Authors' clinical and legal practices; (2) Burn center at regional Level 1 trauma hospital. SUBJECTS: : (1) Case series of face-first, inflicted immersion burn victims; (2) Consecutive hospitalized scald burn victims younger than 5 years old, 1/3/1996 to 3/25/2000. METHODS: (1) Individual case reports; (2) Retrospective records review. Simple descriptive statistics, Fisher Exact test and t test. RESULTS: (1) Six cases of inflicted head and neck immersion injury are described. Four were tap water and 2 food/drink scalds. (2) 22/195 hospitalized victims had sustained immersion burns, 13 from tap water and 9 from other fluids. Six (46%) tap water immersions and no (0%) other immersions had inflicted injuries (P = 0.05). Two of the tap water immersions and one other source immersion included burning of the head and neck. Of these, one tap water immersion, but no other immersion, was inflicted. In no patients were head and neck injuries the sole or predominant site of scalding. In all, 9 children sustained inflicted scalds. Bilateral lower extremity tap water immersion scalds occurred in 100% (6/6) of abusive and 29% (2/7) of unintentional injuries (P = 0.02). Buttock and perineal injuries occurred in 67% (4/6) inflicted versus 29% (2/7) unintentional tap water immersion scalds (P = 0.28). Other fluids caused bilateral lower extremity immersion burns in 3/9 (33 %) unintentionally injured patients, but no abused children (NS). CONCLUSIONS: Craniofacial immersion injury, although seen by the authors in legal cases, is infrequent. It was present incidentally in one inflicted tap water burn in the consecutive hospital series. This series affirms the predominance of bilateral lower extremity burns in inflicted tap water immersions. Buttock/perineal immersions were more common with abuse than with unintentional injury. PMID- 15123902 TI - Surgical emphysema: a rare presentation of foreign body inhalation. AB - An 11-year-old girl with an almond lodging in the tracheobronchial tree is described. She presented with an uncommon symptom of subcutaneous emphysema The x ray revealed left-sided pneumothorax and pneumomediastinum. Intercostal drain was inserted, but she developed respiratory failure and was ventilated. After initial stabilization for 60 hours, she deteriorated again and her x-ray revealed right sided collapse. After removal of the foreign body, she was discharged but presented again with stridor necessitating tracheostomy. Tracheal stenosis was found and required end-to-end anastomosis. The authors feel that, while foreign bodies are uncommon in this age group with emphysema as a rarer manifestation, this cause should be kept in mind, even in the absence of forthcoming history. A high index of suspicion for tracheobronchial foreign body is required in atypical presentations of acute pediatric respiratory distress. PMID- 15123903 TI - Sildenafil citrate ingestion in a pediatric patient. AB - BACKGROUND: Sildenafil citrate is the first FDA-approved oral agent for male erectile dysfunction. Common adverse effects include flushing, headache, and dyspepsia, although more serious side effects have been reported. Because of its specific therapeutic indication, sildenafil toxicity has been limited almost exclusively to adults. We report a symptomatic case of pediatric sildenafil ingestion. CASE: A 2-year-old male ingested 75 mg of sildenafil citrate (Viagra) 2 hours prior to arrival at an emergency room. Ipecac syrup had been given at home with one episode of vomiting. Activated charcoal was considered but withheld due to the delayed presentation to the hospital. The patient was observed in the hospital for 17.5 hours. Observed clinical effects included facial flushing, transient penile engorgement, bilateral rhonchi, and diarrhea. No significant cardiovascular effects were seen. A bronchodilator was given with resolution of rhonchi. No other specific interventions were required. One day after discharge, the patient had one additional bout of diarrhea and complained of pain in the penile region for one day. Two weeks after the exposure, the patient's mother denied any unusual symptoms. CONCLUSION: Pediatric ingestion of sildenafil may result in mild symptoms including persistent flushing and penile engorgement with associated pain. Penile pain may persist even after resolution of the erection. It is questionable whether the respiratory symptoms and diarrhea were related since neither has been described following sildenafil exposure. Significant cardiovascular symptoms were not seen. Early administration of ipecac syrup did not prevent symptoms from developing. PMID- 15123904 TI - Subgaleal hematoma from hair braiding leads to the diagnosis of von Willebrand disease. AB - A 17-year-old male developed a swollen painful scalp hematoma after having his hair braided. This extensive hematoma was drained, but then reaccumulated. Because of this unusual complication, laboratory testing for a bleeding disorder was performed. This patient was diagnosed with type 1 von Willebrand disease. To our knowledge, this is the first case of von Willebrand disease presenting as a subgaleal hematoma. von Willebrand disease is a common inherited bleeding disorder which should be considered in patients with unusual bleeding. PMID- 15123905 TI - Reversal of an antihistamine-induced coma with flumazenil. AB - Flumazenil is a competitive antagonist with specific action at the central benzodiazepine receptor. It is used when benzodiazepine intoxication is suspected. Its use has also been reported in cannabis intoxication, chloral hydrate overdose, hepatic encephalopathy, and alcohol intoxication. We report the case of a 7-month-old male infant with a depressed level of consciousness after intentional intoxication of antihistamines, whose mental status fully recovered after administration of flumazenil. To our knowledge, this is the first case in children where flumazenil has been reported to reverse antihistamine-induced coma. PMID- 15123906 TI - An unusual pediatric hand fracture. PMID- 15123907 TI - Cardiac troponins in pediatrics. PMID- 15123909 TI - Pediatric emergency medicine: legal briefs. PMID- 15123910 TI - Pediatric rapid sequence intubation: a review. AB - Pediatric rapid sequence intubation is a skill of great importance to emergency medicine physicians. Developing a systematic strategy for approaching an emergent airway aids in the proper handling of this event. This paper is a review of the current recommendations for pediatric rapid sequence intubation including current medications and surgical rescue techniques. PMID- 15123911 TI - ECGs in the ED. PMID- 15123912 TI - Changes to EMTALA rules affect pediatric emergency departments. PMID- 15123917 TI - Editorial comment. Computers in orthopaedic surgery. PMID- 15123918 TI - Application of punched card indexing systems to orthopaedic surgery. 1957. PMID- 15123919 TI - A comparison of digital cameras: features essential for the orthopaedic surgeon. AB - To make recommendations for the selection of digital imaging equipment and its use in the typical orthopaedic surgery practice, we investigated four digital cameras with maximal resolution ranging from 1.3-3.34 megapixels. We took images of a plain radiograph, a magnetic resonance image, a hand model, and a minifragment plate with all four digital cameras and a 35-mm film camera. Several variables were evaluated to determine their effect on image quality, including adjusting maximal camera resolution, using a flash, using a camera stand, and using a macromode. Images were graded on a 3-point scale on the computer desktop and as 5 x 7 inch prints by two blinded observers. A maximal camera resolution of 2.1 megapixels was required to make photograph-quality 5 x 7 inch prints of all subjects imaged. No difference in print quality was seen between images taken in the highest quality JPEG format and the uncompressed TIFF format. A macromode with closest focal length less than 5 inches was needed for imaging small subjects. The 1.3-megapixel camera was comparable with the higher resolution cameras for imaging radiographic studies and when viewing color images on the computer desktop. PMID- 15123920 TI - Digital video. AB - The process of digital capture, editing, and archiving video has become an important aspect of documenting arthroscopic surgery. Recording the arthroscopic findings before and after surgery is an essential part of the patient's medical record. The hardware and software has become more reasonable to purchase, but the learning curve to master the software is steep. Digital video is captured at the time of arthroscopy to a hard disk, and written to a CD at the end of the operative procedure. The process of obtaining video of open procedures is more complex. Outside video of the procedure is recorded on digital tape with a digital video camera. The camera must be plugged into a computer to capture the video on the hard disk. Adobe Premiere software is used to edit the video and render the finished video to the hard drive. This finished video is burned onto a CD. We outline the choice of computer hardware and software for the manipulation of digital video. The techniques of backup and archiving the completed projects and files also are outlined. The uses of digital video for education and the formats that can be used in PowerPoint presentations are discussed. PMID- 15123921 TI - Living with digital imaging. AB - Increasingly orthopaedic departments are using digital radiology and imaging systems. We review the technical features of a digital radiology system for the orthopaedic surgeon. The pros and cons of digital and computed radiology are discussed. Hardware and software requirements for storage, retrieval, manipulation, and display of digital images are reviewed. The potential benefits to the clinician and the patient are outlined. PMID- 15123922 TI - Image catalogs. AB - The advent of digital photography and radiography allows documentation of interesting clinical findings with unprecedented ease, and many orthopaedic surgeons have taken extensive advantage of this opportunity to create large digital libraries of clinical results. However, this leaves surgeons with a rapidly increasing volume of data to store and organize; therefore, a system for archiving, locating, and managing images, radiographs, and digital slide presentations has become a crucial need in most orthopaedic groups and practices. However, many surgical groups and practices are not familiar with the computer technology available to initiate such systems. In this review, we discuss several software solutions currently on the market to address the specific needs of orthopaedic surgeons, and as a practical example, discuss a system that is in place in the Department of Orthopaedic Surgery at our institution. Overall, depending on the individual circumstances of each institution, there are various options that meet different technologic and financial requirements. PMID- 15123923 TI - Database production and maintenance. AB - The advent of computers and electronic databases have revolutionized the way that clinical research is done. If study inclusion criteria are saved in the database, creating a study population can be accomplished by constructing an electronic query. Prospectively-entered outcome data related to a particular patient population then can be accessed and exported to a statistical software application for analysis. The senior author has more than 2 decades of experience with computer-based clinical outcome databases. In the context of his personal experience, this review offers some insights related to database design and maintenance. PMID- 15123924 TI - Searching the medical literature. AB - Medical literature searches have become more complex because of the increasing amount of published material and the multiple available databases indexing those publications. Although newly graduated physicians may have received some training in literature searching as part of a medical school curriculum, most clinicians have received no formal training in this skill. In today's world of evidence based medicine, access to published data is crucial, and the importance of a systematic approach to searching cannot be overemphasized. A brief review of searching techniques can greatly increase productivity while using the MEDLINE, Excerpta Medica, Science Citation Index, and other databases. Similarly, knowing the options available for remote access to databases and electronic delivery of articles can expedite the task of accessing publications. PMID- 15123925 TI - Patient education through the Internet: academic and private practice sites. AB - Although Internet use among patients continues to increase, the quality of disseminated information in orthopaedic surgery often is substandard. We reviewed and compared the web sites of academic orthopaedic surgery departments and private practices and rated their informational content with respect to patient education. Only 11.5% of academic sites and 52% of private practice sites list information on common orthopaedic conditions. Of these, less than 1/3 have information on various topics, and few have links to other patient education sites. Private practice web sites make better use of the Internet to provide patients with practical information about their individual practices. Few sites post the date of last update and those that do often have not been updated for more than 6 months. Academic and private practices underutilize the Internet as an educational resource. Orthopaedic surgeons are missing an important opportunity to provide accurate and reliable information to their patients. PMID- 15123926 TI - The digital divide phenomenon in a hand surgery outpatient clinic. AB - The Internet has become an important source of medical information for patients. However, the availability of this resource for individuals in the lower socioeconomic groups is limited, a concept termed the digital divide. To evaluate this phenomenon, we conducted a survey study to quantify the accessibility and use of the Internet for obtaining medical information in an outpatient hand surgery clinic population. A 28-question survey was distributed to 207 patients concerning computer accessibility, Internet use for medical and nonmedical information, Internet trust and security, and patient economic demographics. After analysis of the data, we found individuals in households with higher incomes had a greater likelihood of owning computers than those in lower income brackets. As income increases, the time spent on the Internet also increases. Another statistically significant trend was that higher income patient households thought that Internet information was trustworthy, secure, and private compared with the lower income brackets. We concluded that clinical settings where the predominant patient population earns less than $18,000 may not benefit or use physician Internet-based services or information. The concept of the digital divide seems to be a real phenomenon in the clinical practice of orthopaedics. PMID- 15123927 TI - Evaluating the quality of Internet-derived information on plantar fasciitis. AB - Patients seeking medical advice increasingly turn to the World Wide Web. To test the hypothesis that Internet-provided medical information often is inaccurate and biased, we selected a common musculoskeletal condition. Three common Internet search engines were used to search "plantar fasciitis" and "plantar fascitis." Combining the first 50 web addresses from each search engine for both phrases and eliminating duplicate sites resulted in a database of 152 websites. Each website then was analyzed for authorship, sponsoring agency, and presence of financial incentive. The informational value score of each site then was graded using a 10 point scale evaluating the following parameters: basic definition of plantar fasciitis, causes and risk factors, clinical symptoms, diagnostic tests, differential diagnosis, conservative and surgical treatment options, and complications. The overall mean information value score for all 152 sites was 3.8 points; 36% of the sites showed financial bias. The highest mean scores among all authorship and sponsorship categories belonged to orthopaedic surgeons and academic institutions with mean scores of 7.1 and 6.1 points, respectively. Sites without obvious financial bias had significantly higher information value scores than sites with financial interests. Healthcare professionals should consider these findings when advising patients. PMID- 15123928 TI - Teleradiology in orthopaedics. AB - Teleradiology is a means of electronically transmitting radiographic image files from one location to another. Technologic advances in digital imaging, telecommunications, digital storage, and viewing technologies have made teleradiology readily available and reasonably affordable. The five components of a teleradiology system include: a sending station, a transmission network, a storage device, a viewing station and, a software package. The advantage of teleradiology is the mobility of digital images. In contrast to plain radiographs that only can be seen in one location at a time, multiple persons who are at different locations can view digital images simultaneously. When applied to orthopaedic trauma applications, when the consulting orthopaedist is at a remote location from the patient, teleradiology has been shown to improve diagnostic accuracy, disposition planning of patients from emergency departments or outlying hospitals, and planning of surgical procedures. These systems also improve the comfort level of consulting orthopaedic surgeons and potentially limit the risk of litigation for incorrect diagnosis. The quality, convenience, and effectiveness of teleradiology systems should improve as the technologies continue to mature. Having radiographic images available on handheld devices, such as cell phones, is likely to be a reality in the near future. PMID- 15123929 TI - Image guidance: fluoroscopic navigation. AB - Computer-assisted orthopaedic surgery slowly is making its way into routine orthopaedic practice. Orthopaedic trauma has long been identified as a potential impact area of this new technology. Early experience with three-dimensional (3D) image-guided surgery was promising, but this particular technique was limited by the inability to update the 3D computer model in the operating room after fracture reduction maneuvers or implant placement. Virtual fluoroscopy, or fluoroscopic navigation, became available in 1999 and has proven to be a more versatile technology for fracture treatment. Fluoroscopic navigation systems allow the surgeon to store multiple intraoperative fluoroscopic images on a computer workstation; the position of special optically-tracked surgical instruments or implants then may be virtually overlaid onto the stored images in multiple planes during implant placement. The ability to update images after fracture manipulation now has expanded the application of computer-assisted surgery to any procedures that traditionally have relied on intraoperative C-arm use. In selected applications, this technology has been shown to decrease operative time and intraoperative radiation exposure. The advantages of the new technique of fluoroscopic navigation and its current use in trauma applications will be discussed. PMID- 15123930 TI - Computed tomography-based navigation for hip, knee, and spine surgery. AB - A review of CT-based orthopaedic navigation is presented with a specific emphasis on arthroplasty for the hip and the knee. Fundamental issues about the laboratory and clinical validation of the applications are addressed. The ability to compute the position and orientation of an acetabular implant using a postoperative CT scan was investigated. Angle deviations relative to known positions were computed with an error of less than 1 degree. Then, the system accuracy for three dimensional reconstruction and registration of two cadaveric pelvis specimens was measured with more than 350 registrations. We observed a maximal inclination error of 5 degrees in 99% of cases and a maximal anteversion error of 5 degrees in 97% of cases. The accuracy of the three-dimensional reconstruction and registration for knee arthroplasty also was measured and computed with an angular accuracy of 0.5 degrees in the AP plane and accuracy of 3 degrees in the lateral plane. A clinical study then was done in 109 cases where 96% of implants were installed with a hip-knee-ankle angle of 180 +/- 3 degrees . Computed tomography based navigation for orthopaedic surgery provides greater accuracy and reproducibility than conventional surgery. As noted by learning curves, software improvements are needed to bring it into daily clinical routine. PMID- 15123931 TI - Word recognition software use in a busy orthopaedic practice. AB - For the past 3 years, our orthopaedic office has used word recognition programs to expedite medical record reporting, operative dictation, and letter writing. The use of these programs has resulted in cost savings, increased efficiency, and an improvement in the quality and the thoroughness of our medical records. The methods whereby we were able to incorporate a word recognition program into our operative dictations and our office notes are discussed and explained. Advantages and disadvantages of different programs were evaluated and discussed. We have been able to use word recognition programs in the daily function of our orthopaedic office although not without some difficulties. These programs have resulted in cost savings and time savings and provided medicolegal protection because of the thoroughness of the documentation. The use of templates allows efficiency comparable with conventional dictation. Word recognition programs can be used effectively in busy orthopaedic offices when combined with note templates. PMID- 15123932 TI - The use of a personal digital assistant in orthopaedic surgical practice. AB - The personal data assistant is a powerful tool enabling data acquisition, analysis, and scheduling. The Palm and Windows Pocket PC Operating Systems are available in various personal data assistants that combine bright screens, ease of use, and compactness. Data that are acquired can be imported into standardized spreadsheets for statistical analysis. Report generation using these data can simplify record keeping, facilitate later research, and decrease secretarial typing time. The use of a forms manager, such as Pendragon Forms, enables rapid creation of personal data assistant forms that interface with numerous computer database programs. We currently use theses programs for data acquisition when patients are seen in the office, in the operating room, and when returning for followup. PMID- 15123933 TI - Internet-based learning. AB - The Internet, with established and newly appearing websites, is becoming a more integral part of orthopaedic education every day. We review some of the well known resources on the Internet (Orthogate, OrthoNet, American Academy of Orthopaedic Surgeons site, Orthopedic Hyperguide, WorldOrtho, Wheeless's Textbook of Orthopaedics, Orthoteers, AO North America site, University of Iowa Virtual Hospital texts, and South Australian Orthopaedic Registrars' Notebook) and reports the results of a survey of 35 orthopaedic surgery residents and 24 attending orthopaedic surgeons on the use of orthopaedic knowledge resources on the World Wide Web. The top three most commonly used online sites by the residents were Wheeless's Textbook of Orthopaedics, the American Academy of Orthopaedics Surgeons website, and Orthopedics Hyperguide. According to the survey, all 35 residents used online resources for learning, giving preference to online textbooks when looking for clinical information and online practice examinations when preparing for the in-training (OITE) or board examinations. Among the attending physicians, 19 (79%) used the Internet. Their most commonly used online sites were the American Academy of Orthopaedics Surgeons website, AO North America site, and Wheeless's Textbook of Orthopaedics. The attending physicians also preferred online textbooks to other online resources. Also provided in this article is a discussion of our idea for a future comprehensive, accurate, and constantly updated orthopaedic online resource that uses already available technology. PMID- 15123934 TI - Adoption of information technology by resident physicians. AB - The Internet represents a technological revolution that is transforming our society. In the healthcare industry, physicians have been typified as slow adopters of information technology. However, young physicians, having been raised in a computer-prevalent society, may be more likely to embrace technology. We attempt to characterize the use and acceptance of the Internet and information technology among resident physicians in a large academic medical center and to assess concerns regarding privacy, security, and credibility of information on the Internet. A 41-question survey was distributed to 150 pediatric, medical, and surgical residents at an urban, academic medical center. One hundred thirty-five residents completed the survey (response rate of 90%). Responses were evaluated and statistical analysis was done. The majority of resident physicians in our survey have adopted the tools of information technology. Ninety-eight percent used the Internet and 96% use e-mail. Two-thirds of the respondents used the Internet for healthcare-related purposes and a similar percentage thought that the Internet has affected their practice of medicine positively. The majority of residents thought that Internet healthcare services such as electronic medical records, peer-support websites, and remote patient monitoring would be beneficial for the healthcare industry. However, they are concerned about the credibility, privacy, and security of health and medical information online. The majority of resident physicians in our institution use Internet and information technology in their practice of medicine. Most think that the Internet will continue to have a beneficial role in the healthcare industry. PMID- 15123935 TI - Internal impingement of the shoulder in flexion. AB - Contact of the rotator cuff to the superior glenoid with the arm in flexion has been described and postulated to be a source of rotator cuff disease. The goals of the current study were to document the existence of internal impingement in flexion arthroscopically and to determine its prevalence in patients with various diseases. Also, we attempted to determine the clinical significance of internal impingement in flexion by investigating the associations between internal impingement in flexion and the preoperative and intraoperative findings. A consecutive case series of 376 patients having arthroscopy of the shoulder were entered prospectively into this study. During arthroscopy, intraarticular lesions were evaluated and the presence of contact of the rotator cuff to the superior glenoid and the degree of flexion making the contact were noted. Statistical analysis was done with two dependent variables defined: the presence of internal impingement in flexion and the flexion degree making internal impingement in flexion. Of the 376 patients, 277 (74%) had internal impingement in flexion and 99 (26%) did not have internal impingement in flexion. There were no statistically significant differences in the prevalence of internal impingement in flexion according to the primary diagnoses. Statistical analysis revealed that the presence of internal impingement in flexion was associated with Type II superior labrum anterior posterior (SLAP) lesions and the presence of internal impingement of the rotator cuff to the superior glenoid in abduction and external rotation. There was a significant relationship between rotator cuff disease and decreasing angle of contact for internal impingement in flexion. This study showed that internal impingement in flexion is common in a cohort of patients having shoulder surgery, with an overall prevalence of 74%, and that internal impingement in flexion may contribute to the development of Type II SLAP lesions and rotator cuff disease. PMID- 15123936 TI - Glenoid wear after shoulder hemiarthroplasty: quantitative radiographic analysis. AB - Symptomatic glenoid arthrosis may limit the long-term success of shoulder hemiarthroplasty in patients who are young and functionally demanding. The principal objective of the current study was to quantify glenoid wear after proximal humeral replacement in young, active subjects. Eight patients, ages 21 to 60 years (mean, 45 years), met inclusion criteria. The mean followup was 43 months. Functional scores for the cohort averaged 60% of age and gender-adjusted healthy subjects (range, 28%-84%). Glenohumeral joint space was measured on serial axillary radiographs using a Microscribe 3-DX digitizing device (measurement accuracy, 0.23 mm). Progressive glenoid wear was found in all eight patients. The mean decrease was 2 mm (range, 1.3-2.8 mm), a 68% decrease in glenohumeral joint space. Glenoid cartilage wear also was correlated with Constant and Murley scores. Patients with residual joint spaces less than 1 mm had a mean score of 50%, compared with a score of 71% for patients with joint spaces greater than 1 mm. There were no correlations between wear and mechanism of injury, duration of symptoms, and prior surgery. This study suggests that glenoid cartilage erosion can be expected routinely after humeral head replacement in young, active individuals, and that such wear may adversely affect function or necessitate conversion to total shoulder arthroplasty. PMID- 15123937 TI - Souter arthroplasty for elbows with severe destruction. AB - One hundred fifty-eight primary Souter elbow arthroplasties were done on 134 patients (121 women) with severe joint destruction (Larsen Grade 5) or large bone defects or both. Joint replacement operations were done at our institution from 1985-1997. The study group comprised 156 joints in 132 patients with rheumatoid arthritis or other variants of chronic inflammatory joint disease, one in a patient with osteoarthritis, and one patient with posttraumatic arthrosis. The mean age of the patients at the time of surgery was 57 years (range, 26-81 years) and the mean disease duration was 27 years (tinge, 2-70 years). Radiographically, severe bone defects were detected in 100 humeri and 134 ulnas. Retentive (snap fit) ulnar components were implanted in 110 joints, and bone grafts were used on 26 humeri and 14 ulnas. Major complications led to five early and 16 late reoperations in 19 patients. Four reoperations were done because of dislocation and eight because of aseptic loosening. One reoperation was done because of early infection and five were done because of late infection. One patient had reoperation because of superficial infection in the bursa olecrani and one triceps tendon rupture also was repaired. One patient had wound repair because of marginal necrosis. In the survival analysis, the cumulative success rate without revision for aseptic loosening at 5 years followup was 97%. Despite the demanding nature of these arthroplasties, the primary results are encouraging. Technically, it is possible to do elbow replacement, even on elbows where the humeral condyles or olecranon or both are missing, if there is sufficient bone left on the diaphyseal areas for primary stem fixation. However, in these extreme cases, the poor general condition of the patient or the difficult soft tissue problems in the elbow region may prove to be a contraindication for joint replacement. PMID- 15123938 TI - Influence of articular geometry on prosthetic wrist stability. AB - Ellipsoid and toroid-shaped articulations for total wrist prostheses were evaluated using computer modeling and laboratory experiments. An ellipsoidal design was found to accommodate greater width of the concave proximal component, resulting in better capture and prosthetic stability than a toroid shape. An ellipsoid articulation also provides greater contact area through the available arc of motion. An ellipsoidal articulation is a reasonable design for total wrist arthroplasty. PMID- 15123939 TI - Hydroxyapatite does not improve the outcome of a bipolar hemiarthroplasty. AB - In a one-surgeon study the clinical and radiographic results of 30 cementless bipolar hip prostheses in 24 patients younger than 55 years were evaluated. Eleven noncoated prostheses (Noncoated Group) and 19 hydroxyapatite-coated prostheses (Hydroxyapatite Group) were compared after a mean followup of 10.4 years. The Harris hip score increased from a preoperative average of 41 points to 76 points at final followup (Noncoated Group, 70 points; Hydroxyapatite Group, 80 points). Thigh or groin pain was present in 15 patients (16 hips) (Noncoated Group, 55%; Hydroxyapatite Group, 53%). Radiographically, subsidence was the major problem at the noncoated prostheses (Noncoated Group, 91%; Hydroxyapatite Group, 5%), whereas osteolysis mainly was seen at the hydroxyapatite-coated prostheses (Noncoated Group, 18%; Hydroxyapatite Group, 89%). The obtained aseptic revision rate of 27% (Noncoated Group, 27%; Hydroxyapatite Group, 26%) is too high to use this implant in young patients. The large amounts of polyethylene wear debris generated by the bipolar system play an important role in this limited success. In the initially well-fixed hydroxyapatite-coated prostheses the sealing effect of a hydroxyapatite coating creates high concentrations of polyethylene in the limited joint space, resulting in massive proximal femoral osteolysis. Consequently, a hydroxyapatite coating introduces a new failure mechanism. Therefore, hydroxyapatite does not improve the outcome of a cementless bipolar hemiarthroplasty in the long-term. PMID- 15123940 TI - Outcomes of total hip arthroplasty are similar for patients with displaced femoral neck fractures and osteoarthritis. AB - This study attempted to evaluate whether total hip arthroplasty for displaced femoral neck fractures had significantly different outcomes when compared with total hip arthroplasty for osteoarthritis. This is a retrospective study of 60 patients who had total hip arthroplasties between 1997 and 2001. Thirty patients (mean age, 79.7 years) had total hip arthroplasties for displaced femoral neck fractures; 30 patients (mean age, 76.9 years) were treated with total hip arthroplasties for osteoarthritis. The same surgeon used the modified lateral approach for all surgeries. All patients had radiographic assessment, physical examination, and evaluation with the Harris hip score. The mean followup was 38 months. The mean Harris hip score for the 25 patients treated with a total hip arthroplasty for a femoral neck fracture was 81 points; the mean hip score for the 27 patients treated with a total hip arthroplasty for osteoarthritis was 87 points. No statistically significant differences between these groups were observed. Patients who were treated with a total hip arthroplasty for a femoral neck fracture did not have increased perioperative morbidity compared with patients who had a total hip arthroplasty for osteoarthritis. This study suggests that the outcomes for total hip arthroplasties in this consecutive series of patients treated for displaced femoral neck fractures and osteoarthritis are comparable. PMID- 15123941 TI - Perioperative morbidity in bilateral one-stage total knee replacements. AB - A retrospective review was done on 501 patients who had bilateral sequential one stage total knee replacements from September 1995 to April 2000 to evaluate perioperative (inhospital) morbidity. One thousand two knee replacements were done with the patients receiving regional anesthesia, on 286 women and 215 men with an average age of 66 years. The average transfusion requirement was 2.8 units of blood per patient. There were no deaths, myocardial infarctions, or cerebrovascular accidents. The mean length of hospital stay was 7.2 days. One hundred forty-four perioperative complications were observed in 109 patients (21.8%). These complications included 27 arrhythmias (5%), one congestive heart failure (0.2%), 65 lower extremity deep venous thromboses (13%), 14 fat emboli (3%), and two pulmonary emboli (0.4%). Other major complications were pneumonia (1%), acute renal failure (0.4%), ileus (2%), and mental status changes (2%). Wound complications included two deep infections (0.4%), three hematomas (0.6%), and five delayed wound healings (0.9%). According to the current study the perioperative morbidity and mortality is acceptable if the procedure is used for selected patients. Patients with significant comorbidities should have a staged bilateral total knee replacement. PMID- 15123942 TI - Higher cumulative revision rate of knee arthroplasties in younger patients with osteoarthritis. AB - This study was designed to test the hypothesis that younger patients treated for osteoarthritis and similar conditions using total knee arthroplasty and unicompartmental knee arthroplasty have a lower implant survival rate when compared with older patients. Previous studies have been done on a small number of patients and only included the younger patients. In many cases patients treated for rheumatoid arthritis have been included in the studies and exceptional survival rates have been reported. The current study compared the cumulative revision rate of the components in 33,251 patients older than 60 years and 2606 patients younger than 60 years treated with total knee arthroplasty or unicompartmental knee arthroplasty for osteoarthritis or similar conditions. Cox regression was used to compare the risk for revision between the two age groups and between gender and the effect of year of operation. The results showed a higher cumulative revision rate for the group of younger patients in all statistical analyses and the risk ratio for revision was significantly lower for the group of older patients. The risk for revision decreased for both groups when considering the year of surgery. This is probably attributable to better implant components and surgical techniques. PMID- 15123943 TI - The accuracy of ultrasound for measurement of mobile- bearing motion. AB - After anterior cruciate ligament-sacrificing total knee replacement, mobile bearings sometimes have paradoxic movement but the implications of such movement on function, wear, and implant survival are not known. To study this potential problem accurate, reliable, and widely available inexpensive tools for in vivo mobile-bearing motion analyses are needed. We developed a method using an 8-MHz ultrasound to analyze mobile-bearing motion and ascertained accuracy, precision, and reliability compared with plain and standard digital radiographs. The anterior rim of the mobile bearing was the target for all methods. The radiographs were taken in a horizontal plane at neutral rotation and incremental external and internal rotations. Five investigators examined four positions of the mobile bearing with all three methods. The accuracy and precision were: ultrasound, 0.7 mm and 0.2 mm; digital radiograph, 0.4 mm and 0.2 mm; and plain radiographs, 0.7 mm and 0.3 mm. The interrater and intrarater reliability ranged between 0.3 to 0.4 mm and 0.1 to 0.2 mm, respectively. The difference between the methods was not significant for neutral rotation but ultrasound was significantly more accurate than any one degree of rotation or higher. Ultrasound of 8 MHz provides an accuracy and reliability that is suitable for evaluation of in vivo meniscal bearing motion. Whether this method or others are sufficiently accurate to detect motion leading to abnormal wear is not known. PMID- 15123944 TI - Eight- to 14-year followup of arthroscopic meniscal repair. AB - The current study aims to elucidate the midterm to long-term progression of arthroscopic meniscal repair which had been confirmed through a second-look arthroscopy at an average of 10 months after repair. Twenty-eight menisci of 28 patients were investigated at 8 to 14 years (mean, 10.2 +/- 1.8 years), eight after isolated meniscal repair (Isolated group) and 20 after concomitant anterior cruciate reconstruction (Anterior cruciate ligament group). All were rated excellent except four that rated good in the anterior cruciate ligament group as indicated by Lysholm's score at followup. None of the Isolated group, but 12 of the Anterior cruciate ligament group, had osteoarthritic changes seen on radiographs. Four and 19, respectively, had increased signal intensity on magnetic resonance imaging scans, 10 in the Anterior cruciate ligament group had changes of Grade 3 severity. However, no meniscal signs or symptoms were present and no significant relationship between findings of radiographs and magnetic resonance imaging scans was apparent. Anterior cruciate reconstructed knees that were changed to Grade 3 severity as seen on magnetic resonance imaging scans had higher sagittal instability seen on stress radiography than knees with Grade 2 or less severe changes. The anterior laxity of the knees influenced the severity of meniscal signal changes on the magnetic resonance imaging scans. PMID- 15123945 TI - The modular segmental kinematic rotating hinge for nonneoplastic limb salvage. AB - From January 1980 to July 1998, 25 patients (26 knees) were treated with an arthroplasty using a Modular Segmental Kinematic Rotating Hinge total knee prosthesis for nonneoplastic limb salvage. The indications included: nonunion of a periprosthetic femur fracture (11 knees), severe bone loss and ligamentous instability (eight knees), nonunion of a supracondylar femur fracture (four knees), acute periprosthetic fracture (one knee), fracture of a previous hinge (one knee), and prior resection arthroplasty (one knee). The average age of the patients was 72.3 years. Twenty-two arthroplasties were revisions. The average followup was 58.5 months. At the latest followup, knee extension averaged 2.4 degrees and flexion averaged 93.6 degrees. The Knee Society knee score improved from an average of 45.4 preoperatively to 75.5. Preoperatively, functional scores averaged 8.6 and improved to 25. Complications occurred in eight patients. The most common was deep infection (five patients). The use of the Modular Kinematic Rotating Hinge for nonneoplastic limb salvage represents a small proportion (0.14%) of all primary and revision knee arthroplasties done at our institution. The indications for the surgery are for a highly complex and small subset of patients. The patients in the current study gained significant improvement in overall range of motion, Knee Society knee scores, and functional scores when this prosthesis was used. PMID- 15123946 TI - Effects of knee pain relief in osteoarthritis on gait and stair-stepping. AB - Osteoarthritic knee pain affects patient mobility. Relief of knee pain in osteoarthritis has been reported to increase loading of the knee during gait, but it is unknown whether such pain relief enhances knee loading during more demanding activities such as stair-stepping. The gait of 19 patients and stair stepping of 14 patients with painful medial compartment osteoarthritis of the knee was assessed before and after pain-relieving intraarticular injection of the knee and compared with those of 21 healthy control subjects. There were significant increases in gait velocity, cadence, maximum external knee adduction moment (indicating increased loading in the medial compartment of the knee), and maximum external hip adduction and ankle abduction moments immediately after the injection. With the exception of velocity and ankle abduction moment, these variables were returned to levels that were not statistically different from those of the control subjects. However, no significant differences were found during stair-stepping in the external adduction-abduction moments about the knee, hip, or ankle after injection. Furthermore, the postinjection magnitudes of these variables during stair-stepping were significantly less than those of the controls. Therefore, although the relief of knee pain is sufficient to enhance gait function in osteoarthritis of the knee, it is insufficient to enhance stair stepping function. PMID- 15123947 TI - Modifications of the Weil osteotomy have no effect on plantar pressure. AB - Previous studies have shown that increasing angulation of the Weil osteotomy produces greater plantar translation of the metatarsal head. Modifications have been proposed to reduce plantar translation. However, there is no evidence that the increased plantar translation with a Weil osteotomy is clinically significant or that these modifications are required. Ten lower extremities consisting of five matched pairs were used to evaluate whether different configurations of the Weil osteotomy altered plantar pressure in a dynamic cadaver model. For each pair, an oblique Weil osteotomy with a 5-mm shift was done on one side and a standard (parallel) Weil osteotomy with a 5-mm shift was done on the matched foot. A 4-mm slice resection and a metatarsal head resection then were done sequentially. Plantar pressures were measured with cyclic loading to 700 N at a frequency of 1 Hz with an F-scan in-shoe sensor on the intact specimens and after each intervention. Increased plantar translation of the metatarsal head with a more oblique Weil osteotomy did not significantly increase plantar pressure, and the 4-mm slice resection did not significantly unload the metatarsal head. Only complete metatarsal head resection significantly unloaded the metatarsal head. PMID- 15123948 TI - Second metatarsophalangeal joint: biomechanics and reconstruction potential. AB - A damaged distal radioulnar joint results in instability, pain, and loss of motion. This amounts to function loss. Of the many proposed reconstructive procedures, only a vascularized joint transfer offers growth potential in children and durability. The well-studied vascular anatomy and minimal donor morbidity of the second metatarsophalangeal joint makes it a potentially useful source. The authors evaluated various biomechanical aspects of the second metatarsophalangeal joint, which included the mediolateral arc of movement of the second metatarsophalangeal joint in hyperextension when subjected to lateral stress loading; the different patterns of disruption at peak load; and the flexion and extension range of movement. Twenty-six cadaveric specimens were tested with a customized jig. The results showed that the mediolateral arc of movement of 114 degrees approximates that of the flexion and extension range of movement of 114.2 degrees. However, it differed in that it was equal in both directions, compared with an extension arc that was greater than a flexion arc. The peak load was approximately 100 N, and this resulted in fracture, avulsion, and ligament tear. These biomechanical results may be useful to reconstructive surgeons. PMID- 15123949 TI - Surgical treatment and recurrence rate of aneurysmal bone cysts in children. AB - High recurrence rates have been reported after surgical treatment of aneurysmal bone cysts, ranging from 10-59%, and greater (five of seven) in children younger than 10 years. No previous study has focused specifically on recurrence as a function of age in a large pediatric study. The purpose of the current study was to review the outcome of current surgical techniques in the treatment of primary aneurysmal bone cysts in children and to determine if recurrence rates were higher in younger children, as proposed by some authors. This study included 45 children with primary aneurysmal bone cysts with more than 2 years followup after their first treatment using our current four-step surgical technique. The subjects were studied and classified on the basis of their age group (< or = 10 years of age or >10 years of age). The recurrence rate, and other parameters were determined after surgical treatment. Of the 45 children with primary aneurysmal bone cysts, 13 children were 10 years or younger and 32 were older than 10 years. Persistence or recurrence of the lesion requiring additional surgery occurred in three children in the younger age group and in five children in the older age group. The difference in persistence or recurrence rates based on age was not statistically significant. However, the persistence or recurrence rates are lower than previously reported (82% overall success rate after initial surgery). PMID- 15123950 TI - Osteosarcoma in children: long-term functional analysis. AB - Multimodal, limb-saving therapy is the treatment of choice in children with osteosarcoma. However, few data are available on long-term functional problems and disabilities in surviving patients. Seventy-eight children treated for Stage II-B osteosarcoma who were treated between 1970 and 1997 were evaluated. Beyond the clinical examination, functional results were assessed using the scoring system of the Musculoskeletal Tumor Society. Patients reported their physical disabilities when completing the Toronto Extremity Salvage Score. The most recent cohort of patients who had tumor resection combined with preoperative and postoperative chemotherapy had the best 10-year survival rate at 70%. Thirty-four children (44%) were treated by limb-sparing therapy with 25 children requiring an endoprosthesis. Ten of these 25 children (40%) required one or in some cases several replacements of the endoprostheses because of complications. The original prostheses were in place for a median of 4.9 years before the first exchange (range, 2.0-12.0 years). Evaluations with rating scales showed that the subjective estimation of disabilities by the Toronto Extremity Salvage Score was significantly higher than a rating of limb functions by a doctor using the Musculoskeletal Tumor Society score. Best results were observed in children with an endoprosthesis of the distal femur who achieved as much as 90% ability to do their daily life routine. Limb-saving therapy in children with osteosarcoma enables a return to activities of daily living in long-term survivors with a minimum of remaining disability. PMID- 15123951 TI - Giant cell tumor and Paget's disease of bone in one family: geographic clustering. AB - Giant cell tumor is a rare complication of Paget's disease of bone. Typically, this tumor occurs in the case of polyostotic disease and only in pagetic bones. This tumor rarely has been seen in multiple family members who have Paget's disease, although Paget's bone disease clearly has a hereditary component. Our report documents four cases of polyostotic Paget's bone disease complicated by benign giant cell tumor. In two patients, the giant cell tumor also was multifocal. All patients were from one family. They were born in Avellino and reside in Campania, a Southern Italian region. The ancestors of the patients with familial giant cell tumor in Paget's bone disease were born in the same geographic area. These data suggest that a combination of environmental and genetic factors could be responsible for linkage of the patients born in Avellino with this neoplasm that is highly unusual in patients with Paget's disease of bone. PMID- 15123952 TI - Leiomyosarcoma of somatic soft tissues. AB - Leiomyosarcoma is a rare, aggressively malignant connective tissue tumor of mature adults, which arises from smooth muscle. It occurs most frequently in the uterus, bowel, vascular tissues, and less commonly in somatic soft tissue or bone. The tumor when it arises in soft tissue has distinctive histologic features which somewhat resemble malignant fibrous histiocytoma (otherwise known as myxofibrosarcoma). The Orthopaedic Oncology Service at our institution has treated 66 patients with these lesions and thus far, 1/2 of the patients have died of disease at a mean of 3 years after discovery. Factors that increase the death rate include size of the tumor, Musculoskeletal Tumor Society Stage of disease, and to a lesser extent particularly in the lower extremities, anatomic site. Radiation and chemotherapy had little direct effect on the outcome but patients treated with surgery and adjunctive agents seemed to live longer than their cohorts treated with surgery alone. The purpose of this study is a general review of the clinical and prognostic features of this cancer. PMID- 15123953 TI - Allografts about the Knee in Young Patients with High-Grade Sarcoma. AB - Reconstruction after resections for high-grade sarcomas about the knee in children and adolescents is a challenging problem because of the large soft tissue and skeletal defects, the effects of adjuvant therapy, and the potential for long-term use of the limb. One hundred sixteen patients, all 18 years or younger, with osteosarcoma or Ewing's sarcoma located between the middle femur and middle tibia, were treated with chemotherapy, resection, and allograft reconstruction. One hundred three patients with osteosarcoma and 13 patients with Ewing's sarcoma had 105 Stage II and 11 Stage III tumors. There were 72 osteoarticular grafts (39 femur, 33 tibia), 28 intercalary grafts (19 femur), seven allograft-prosthetic composites (all femur,) and nine allograft-arthrodeses (seven femur, two tibia). At latest followup, 49% of all of the allograft reconstructions were rated good or excellent, 14% were rated as fair, and 37% were failures. Sixteen percent had an infection develop. Twenty-seven percent of patients had a fracture, 34% had a nonunion, and 14 patients eventually required amputation. Reconstruction of large bone defects about the knee in young patients who are being treated with chemotherapy is difficult. Although complications significantly affect outcome, allografts are a viable option for reconstruction in children with high-grade sarcomas about the knee. PMID- 15123954 TI - Operative treatment of tibial plateau fractures in patients older than 55 years. AB - Surgical treatment of tibial plateau fractures in the older patient poses an additional challenge because of the underlying condition of the bone and articular surface. We sought to identify risk factors for poorer outcomes in the operative treatment of displaced tibial plateau fractures in older patients. Thirty-nine displaced tibial plateau fractures in patients 55 years and older were treated operatively. Patients were evaluated objectively with Rasmussen clinical and radiologic scoring techniques, and the Short Musculoskeletal Function Assessment and the Short-Form 36 self-assessment instruments. The Rasmussen clinical and radiologic scoring systems, used on average 2.54 years postoperatively, found acceptable results in 87.2% and 82.1% of patients, respectively. The fracture classification of Schatzker was not predictive of results. External fixation was associated with significantly poorer results. Increasing age was associated with poorer clinical and self-assessment scores, although preexisting degenerative joint disease was not. The results from the Short-Form 36 indices were not significantly worse for our study patients. The average Short Musculoskeletal Function Assessment score of our study patients indicated poorer function for mobility than a normative group. Operative treatment of this injury in this population can result in favorable outcomes as evaluated by clinical, radiographic, and self-assessment criteria. PMID- 15123955 TI - Undisplaced intracapsular hip fractures: results of internal fixation in 375 patients. AB - Three hundred seventy-five patients with an undisplaced intracapsular proximal femoral fracture were treated with internal fixation. Nonunion occurred in 24 patients (6.4%) and avascular necrosis occurred in 15 patients (4.0%). Reoperation with an arthroplasty was required in 29 patients (7.7%). The age, walking ability of the patient, and degree of impaction seen on the anteroposterior radiograph or angulation seen on the lateral radiographs were of statistical significance in predicting fracture healing complications. The results for this series of patients were compared with the results in published reports identified by a comprehensive literature search. Summation of the results indicated that the overall risk of redisplacement or nonunion of the fracture was 4.3% (95% confidence interval, 3.4%-5.3%) with internal fixation of an undisplaced intracapsular fracture. For conservative treatment, the failure rate was 19.6% (95% confidence interval, 17.2%-22.1%). The incidence of avascular necrosis with internal fixation at 1 year was 2.2% (95% confidence interval, 1.6% 2.9%) compared with 2.8% (95% confidence interval, 1.9%-4.0%) with nonoperative treatment. Internal fixation is recommended for the treatment of undisplaced intracapsular hip fractures. PMID- 15123956 TI - Blastomycosis of long bones. AB - The presentation of blastomycosis clinically and radiographically is nonspecific and often mistaken for a neoplasm. Delay in diagnosis is common. Patients with osseous blastomycosis present with pain and swelling. Radiographs usually show an eccentric lucency in the distal ends of long bones. These patients frequently are referred for a neoplastic workup and a diagnosis is made only after biopsy. We review the cases of five patients diagnosed with a bone tumor who had blastomycosis osteomyelitis. The time to diagnosis from original symptoms was 4.7 months (range, 3-8 months). The average age of the patients was 45.6 years (range, 20-59 years). A Musculoskeletal Tumor Society functional assessment was done. Early radiographs of the current patients ranged from normal to showing faint osteopenia in the involved location. As the disease progressed, the area of lucency appeared with either diffuse or well-marginated borders. Treatment included surgical debridement with antifungals. The mean functional score was 93.3%. All patients are disease-free. Blastomycosis, similar to tuberculosis, often is mistaken for a neoplasm. Blastomycosis osteomyelitis can be treated with excellent results. The key is diagnosis and including endemic fungal infections in the differential diagnosis of bone tumors. In addition, every potential neoplasm should include cultures of specimens obtained at biopsy. PMID- 15123957 TI - Magnetic resonance imaging in stress fractures and shin splints. AB - The purpose of the current study was to determine whether stress fractures and shin splints could be discriminated with MRI in the early phase. Twenty-two athletes, who had pain in the middle or distal part of their leg during or after sports activity, were evaluated with radiographs and MRI scans. Stress fractures were diagnosed when consecutive radiographs showed local periosteal reaction or a fracture line, and shin splints were diagnosed in all the other cases. In all eight patients with stress fractures, an abnormally wide high signal in the localized bone marrow was the most detectable in the coronal fat-suppressed MRI scan. In 11 patients with shin splints, the coronal fat-suppressed MRI scans showed a linear abnormally high signal along the medial posterior surface of the tibia, and in seven patients with shin splints, the MRI scans showed a linear abnormally high signal along the medial bone marrow. No MRI scans of shin splints showed an abnormally wide high signal in the bone marrow as observed on MRI scans of stress fractures. This study showed that fat-suppressed MRI is useful for discrimination between stress fracture and shin splints before radiographs show a detectable periosteal reaction in the tibia. PMID- 15123958 TI - Cryotherapy decreases intraarticular temperature after ACL reconstruction. AB - Cryotherapy has been shown to decrease intraarticular temperature in all regions of the knee after arthroscopy. The purpose of our study was to determine if similar declines in intraarticular temperature were seen with the use of cryotherapy after anterior cruciate ligament reconstruction, a procedure which, unlike simple arthroscopy, produces postoperative hemarthrosis. Sixteen patients had intraarticular temperatures measured for 2 hours after anterior cruciate ligament reconstruction with cryotherapy instituted for only 1 of the 2 hours (eight patients for the first hour and eight patients for the second hour). Significant declines were seen in the suprapatellar pouch after either hour (-2.7 degrees C in Group 1, -2.7 degrees C in Group 2) but not in the lateral gutter. The difference between cryotherapy versus no cryotherapy in the first hour in the suprapatellar pouch was 6.0 degrees C, a clinically meaningful temperature difference. We hypothesize the swelling and hemarthrosis was more pronounced in the lateral gutter because of its dependent position and therefore blunted the effect of cryotherapy seen in the lateral gutter. Cooling of the intraarticular temperature should be considered in the clinical benefits of cryotherapy. PMID- 15123959 TI - Histologic findings of anterior cruciate ligament reconstruction with Achilles allograft. AB - The purpose of the current study was to evaluate the remodeling process of Achilles allograft for anterior cruciate ligament reconstruction under light microscopic and electron microscopic evaluations. In 12 patients, histologic examinations were done preoperatively and during arthroscopy at 6, 12, and 24 months. Gross observations were made with respect to width, vascularity, elasticity, and stability of the Achilles allograft. Biopsy specimens were obtained at the followup arthroscopic study. Light microscopic examinations done at 6 months showed many fibroblasts with spindle-shaped nuclei and hypercellularity. At 12 months, the small folds pattern of collagen bundles and decreased cellularity were observed. At 24 months, gross and findings of the light microscopic evaluations of the allograft tendons were similar to those of the normal anterior cruciate ligaments. On electron microscopic examinations done at 24 months, the allograft had fibroblasts with much cytoplasm and densely packed parallel-laid collagen fibrils, which showed the characteristic cross striations, but the Achilles allograft ligaments did not show similar findings compared with biopsy samples from normal anterior cruciate ligaments. PMID- 15123960 TI - Triamcinolone suppresses human tenocyte cellular activity and collagen synthesis. AB - Glucocorticoid injection is widely used in tendon disorders. Despite previous studies on the histologic and biomechanical changes in tendons after glucocorticoid injections, the role of glucocorticoid in tendon rupture still is controversial. It was hypothesized that glucocorticoid has a direct deleterious effect on human tenocytes, suppressing its cellular activity and collagen production. Primary cultures of human tenocytes were obtained from explants of healthy patellar tendon harvested during anterior cruciate ligament reconstructions. The effects on cell viability and cell proliferation were measured by [3-(4,5-demethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay and 5-bromo-deoxyuridine incorporations. The effect on collagen synthesis was measured by H-proline incorporation assay. Triamcinolone acetonide at 10 to 10 mol/L decreased human tenocyte viability to 45% to 88% of control in a dose dependent manner. Cell proliferation was suppressed to 87% +/- 8% at all doses. Treatment with 1 micromol/L triamcinolone acetonide reduced the amount of collagen synthesis as measured by H-proline incorporation from 40 +/- 2 cpm/1000 cells to 27 +/- 4 cpm/1000 cells. The suppressed human tenocyte cellular activity and reduced collagen production may lead to disturbed tendon structure and predispose the tendon to subsequent spontaneous rupture. PMID- 15123961 TI - How accurate is partial weightbearing? AB - The accuracy of partial weightbearing was assessed in six healthy volunteers and 23 patients who had sustained either a fracture of a lower limb or surgery. They were trained to weightbear partially using the conventional bathroom scale method and were assessed in a gait laboratory using force platforms. The amount of weight exerted on the involved limb during three-point crutch walking was determined. Four of six volunteers exerted a mean of 27% of body weight more than required. The remaining two volunteers exerted a mean of 8.5% of body weight less than required. Of the 23 patients, 21 exerted a mean of 35.3% of body weight more than that prescribed and two patients exerted a mean of 11.97% of body weight less than that prescribed. In both groups there was little relationship between the weightbearing prescribed and actual weightbearing. None of the patients or volunteers was able to reproduce the extent of partial weightbearing for which they were trained using the bathroom scale method, confirming that this technique of instructing patients in partial weightbearing is inaccurate. PMID- 15123962 TI - Asymmetric limb loading with true or simulated leg-length differences. AB - Unequal leg lengths result in asymmetric limb loading but opinions vary on the size of the difference inducing abnormal loading, and which limb sustains the greater load. Our study compared limb-loading asymmetries during walking for subjects with anatomic leg-length discrepancies between 1.0 and 3 cm, subjects without length discrepancies, and for subjects with a simulated a 1.31-cm leg length discrepancy. Symmetry indices were calculated for peak ground reaction force during weight acceptance, rate of change of weight acceptance force, peak push-off force, and rate of change of push-off force. All symmetry measures were significantly different from normal for the simulated leg-length discrepancy. The shorter limb sustained a greater proportion of the load and loading rate. The anatomic leg-length discrepancy group showed the same trend with the exception of the push-off force rate. There were equivalent size-effect differences for both leg-length discrepancy conditions; however, for the anatomic leg-length discrepancy group, only the weight acceptance force symmetry value was statistically different from normal. The shorter limb sustains a greater proportion of load and loading rates; therefore, equalizing leg lengths should be considered even with bilateral differences less than 3 cm. PMID- 15123963 TI - Effective treatment of osteomyelitis with biodegradable microspheres in a rabbit model. AB - Biodegradable microspheres were manufactured from a high molecular weight copolymer of 50% lactic and 50% glycolic acid and the antibiotic tobramycin. It was hypothesized that the microspheres would be more effective than polymethylmethacrylate beads in the local delivery of tobramycin and that the microspheres would not inhibit bone healing. Osteomyelitis was established in 40 New Zealand White rabbits using Staphylococcus aureus. All animals had irrigation and debridement of the infected radii four weeks after inoculation and were divided into five treatment groups: debridement alone, microspheres alone, microspheres containing tobramycin plus parenteral treatment with cefazolin, polymethylmethacrylate beads containing tobramycin plus parenteral cefazolin, and parenteral cefazolin. All animals were sacrificed after 4 weeks of treatment. The group treated with microspheres plus parenteral antibiotics was the only group to have a significantly higher percentage of animals without bacteria after 4 weeks of treatment when compared with the control group. Additionally, the animals treated with microspheres had a higher degree of bone healing in the defect than the animals treated with bone cement. The most effective treatment was biodegradable microspheres combined with parenteral antibiotic in this rabbit osteomyelitis model. PMID- 15123964 TI - Thigh mass in a 9-year-old boy. PMID- 15123965 TI - Sternal pain in an 11-year-old boy. PMID- 15123966 TI - A 36-year-old woman with a toe mass. PMID- 15123968 TI - Hospitals are nursing institutions. PMID- 15123969 TI - Nursing care of the patient with preterm premature rupture of membranes. AB - Preterm premature rupture of the membranes (PPROM) is diagnosed when rupture of the amniotic membranes occurs prior to the completion of the 36th week of gestation. PPROM accounts for 25% of all cases of premature rupture of the membranes and is responsible for 30%-40% of all preterm deliveries. In mothers diagnosed with PPROM without evidence of infection, active labor, or fetal compromise, the current standard of care is expectant management. The goal of expectant management is the prolongation of the pregnancy to increase fetal gestational age thus potentially decreasing the effects of prematurity. Expectant management consists of ongoing observation for signs and symptoms of infection, active labor, and/or nonreassuring fetal status. This article provides clinical nursing guidelines for the mother diagnosed with PPROM who is managed expectantly. Eight targeted areas for nursing assessment and intervention are described: preterm labor, side effects of tocolytic therapy, maternal/fetal infection, fetal compromise, side effects of extended bed rest, maternal stress, educational needs, and routine prenatal care. PMID- 15123970 TI - The efficacy of facilitated tucking for relieving procedural pain of endotracheal suctioning in very low birthweight infants. AB - PURPOSE: This study compared the efficacy of a behavioral pain reducing intervention (facilitated tucking) with standard neonatal intensive care unit (NICU) care for decreasing procedural pain (endotracheal suctioning) in very low birthweight (VLBW) infants. STUDY DESIGN AND METHODS: A prospective randomized crossover design with infants as their own controls were used. The sample consisted of 40 VLBW infants, 23-32 weeks gestation, and weighing 560-1498 g with tracheal intubation. The infants were observed twice during each endotracheal suctioning experience; one suctioning was done according to normal nursery routine; another was done using facilitated tucking (the caregiver "hand swaddling" the infant by placing a hand on the infant's head and feet while providing flexion and containment). The Premature Infant Pain Profile (PIPP) measured the infant's pain response, and severity of illness of each infant was measured by the Score for Neonatal Acute Physiology (SNAP) and the NTISS (Neonatal Therapeutic Intervention Scoring System). Repeated measures analysis of variance (RMANOVA) determined the efficacy of facilitated tucking for reducing procedural pain (PIPP) and the effects of order of intervention vs. control. Regression analyses examined the relationship of gestational age, severity of illness, and number of painful procedures to the pain response. RESULTS: There was a significant difference between the PIPP scores for tucking and nontucking positions (p = 0.001) and a nonsignificant interaction with order (p = 0.64) as well as a nonsignificant main effect for order (p = 0.46). In the regression analyses, all predictors taken together did not significantly predict PIPP scores in the tucked position (p = 0.11) or nontucked position (p = 0.57). CLINICAL IMPLICATIONS: Facilitated tucking is a developmentally sensitive, nonpharmacological comfort measure that can relieve procedural pain in VLBW infants. Nurses need to be increasingly aware of infant pain during daily care taking, and to use validated pain assessment instruments. Further clinical research on individual pain assessment is needed for better understanding of the quality and significance of pain for each infant, and the factors that affect pain expression. PMID- 15123972 TI - Obstetrical accidents involving intravenous magnesium sulfate: recommendations to promote patient safety. AB - Magnesium sulfate is commonly used in obstetrical practice both as seizure prophylaxis in women with preeclampsia, as well as to inhibit preterm labor contractions. However, despite (and perhaps because of) years of use and provider familiarity, the administration of magnesium sulfate occasionally results in accidental overdose and patient harm. Fortunately, in most instances when potentially fatal amounts of magnesium sulfate are given, the error is recognized before permanent adverse outcomes occur. Nevertheless, a significant and sometimes unappreciated risk of harm to mothers and babies continues to exist. Intravenous magnesium sulfate treatment has become routine practice in obstetrics, but this does not lessen the vigilance required for safe care for mothers and babies. Implementation of the recommendations provided in this article will promote patient safety and decrease the likelihood of an accidental overdose, as well as increase the chances of identifying an error before a significant adverse outcome occurs. PMID- 15123974 TI - Are nurses acting as role models for the prevention of SIDS. AB - PURPOSE: To examine nurses' knowledge, attitude, and practice in positioning healthy newborns for sleep in the hospital setting. DESIGN AND METHODS: A cross sectional descriptive design was used to survey a convenience sample of practicing maternal child nurses in 58 Missouri hospitals. A 24-item investigator designed questionnaire was developed with input from SIDS Resources in Missouri. RESULTS: A total of 528 surveys were analyzed. These nurses reported no longer placing infants in the prone position for sleep, but almost 75% of those answering the survey used either the side-lying position or a mixture of side and back positioning, even though 96% of the nurses said they were aware of the AAP Guidelines recommending "back to sleep." Forty-five percent of the nurses thought the infant would be at risk for aspiration if only placed on his/her back. Only 53% of the nurses knew their hospital's policy about newborn positioning; 80% of those who knew about the policy said it included the lateral position as being acceptable practice. CLINICAL IMPLICATIONS: Nurses are the role models for new parents regarding newborn sleep position, and are in a unique position to influence parents' decisions about how to place their infants for sleep at home. Because nurses continue to worry about aspiration when newborns are placed on their backs, it is clear that more education is needed for hospital nurses about newborn sleep position and hospital policies, as well as AAP Guidelines. PMID- 15123975 TI - Postpartum depression as profiled through the depression screening scale. AB - PURPOSE: To describe in greater depth the profiles of the dimensions of the Postpartum Depression Screening Scale (PDSS), with a focus on those women identified with postpartum depression (PPD), and to provide a discussion of the implications for early detection in the community. METHOD AND DESIGN: Subanalysis of data from 150 new mothers who completed the PDSS and had a DSM-IV diagnostic interview conducted by a nurse psychotherapist. Data analysis focused on exploring the profiles of women who were diagnosed with PPD as well as those who were not. RESULTS: The respondents within the major PPD group averaged scores twice as high as those in the nondepressed group, and described more profound emotional responses to their maternal role transitions. CLINICAL IMPLICATIONS: PPD should be conceptualized as occurring in a continuum, with symptoms worsening over time for some women. Nurses working across healthcare settings can use the PDSS for identifying women with PPD. All healthcare providers who are in contact with postpartum women should be open to discussing these women's emotional needs, and should assess women on an ongoing basis. Using all opportunities to listen to women's unique stories that unfold during the postpartum period can help identify women who require treatment for this curable illness. PMID- 15123976 TI - Deep vein thrombosis in pregnancy. AB - This article provides a review of the incidence, pathophysiology, and treatment of deep vein thrombosis (DVT) in pregnancy, a rare but serious complication of pregnancy. The incidence of DVT in pregnancy varies widely, but it is a leading cause of maternal morbidity in both the United States and the United Kingdom. Risk factors during pregnancy include prolonged bed rest or immobility, pelvic or leg trauma, and obesity. Additional risk factors are preeclampsia, Cesarean section, instrument-assisted delivery, hemorrhage, multiparity, varicose veins, a previous history of a thromboembolic event, and hereditary or acquired thrombophilias such as Factor V Leiden. Heparin is the anticoagulant of choice to treat active thromboembolic disease or to administer for thromboprophylaxis, but low molecular-weight heparin is being used with increasing frequency in the pregnant woman. Perinatal nurses should be aware of the symptoms, diagnostic tools, and treatment options available to manage active thrombosis during pregnancy and in the intrapartum and postpartum periods. PMID- 15123977 TI - Dimensions of women's long-term postabortion experience. AB - PURPOSE: To explore and describe the long-term postabortion experience as lived by women, at least 5 years after a first-trimester-induced abortion. METHODS: This phenomenological study used semistructured interviews and constant comparison analysis. Stories of 17 women were recorded on audio tape, transcribed verbatim, coded, and analyzed. RESULTS: Women who had induced abortions represented several ethnic groups, religions, and occupations, and were single, married, or divorced. The average number of years from abortion to interview was 18.9 (range 6-31 years). The age range at first abortion was 14 to 43, and at interview was 23 to 60 years of age. Five themes emerged within the women's stories: Making the Decision, Coping With the Memories, Gaining Perspective, Seeking Help, and Recognizing Its Worth. Most women who participated in this study were able to integrate the abortion experience into their lives, and had found meaning in the abortion experience. CLINICAL IMPLICATIONS: This study provides yet more reasons why nursing should encourage women to prevent unplanned pregnancies through fertility control. Women in this study described many life changing experiences, both positive and negative, because of an abortion. Therefore, preabortion counseling should be sensitive and include information about possible long-term effects. Postabortion support should acknowledge spiritual issues, and include steps women can take to help heal themselves, such as grief counseling and mourning rituals when appropriate. PMID- 15123978 TI - Thiamine (vitamin B1) revisited. PMID- 15123979 TI - Evaluating the nursing PDA discussion list. PMID- 15123981 TI - Terbutaline and methergine in look-alike packaging. PMID- 15123982 TI - Assisted reproductive technology surveillance--United States, 2001. AB - PROBLEM/CONDITION: In 1996, CDC initiated data collection regarding assisted reproductive technology (ART) procedures performed in the United States to determine medical center-specific pregnancy success rates, as mandated by the Fertility Clinic Success Rate and Certification Act (FCSRCA) (Public Law 102-493, October 24, 1992). ART includes fertility treatments in which both eggs and sperm are handled in the laboratory (i.e., in vitro fertilization and related procedures). Patients who undergo ART treatments are more likely to deliver multiple-birth infants than women who conceive naturally. Multiple births are associated with increased risk for mothers and infants (e.g., pregnancy complications, premature delivery, low-birthweight infants, and long-term disability among infants). REPORTING PERIOD: 2001. DESCRIPTION OF SYSTEM: CDC contracts with a professional society, the Society for Assisted Reproductive Technology (SART), to obtain data from fertility medical centers located in the United States. Since 1997, CDC has compiled data related to ART procedures. The Assisted Reproductive Technology Surveillance System was initiated by CDC in collaboration with the American Society for Reproductive Medicine, the Society for Assisted Reproductive Technology, and RESOLVE: The National Infertility Association. RESULTS: In 2001, a total of 29,344 live-birth deliveries and 40,687 infants resulting from 107,587 ART procedures were reported from 384 medical centers in the United States and U.S. territories. Nationally, 80,864 (75%) of ART treatments used freshly fertilized embryos from the patient's eggs; 14,705 (14%) used thawed embryos from the patient's eggs; 8,592 (8%) used freshly fertilized embryos from donor eggs; and 3,426 (3%) used thawed embryos from donor eggs. Overall, 40% of ART procedures that progressed to the transfer stage resulted in a pregnancy; 33% resulted in a live-birth delivery (delivery of > or =1 infant); and 21% resulted in a singleton live birth. The highest live-birth rates were observed among ART procedures using freshly fertilized embryos from donor eggs (47%). The greatest numbers of ART procedures were performed among residents of California (13,124), New York (12,379), Massachusetts (8,151), Illinois (7,933), and New Jersey (6,011). These five states also reported the highest number of live-birth deliveries and infants born as a result of ART. The ratio of number of ART procedures per million population ranged from 74 in Idaho to 1,273 in Massachusetts, with a national average of 371 ART procedures started per million persons. Among ART treatments in which freshly fertilized embryos from the patient's eggs were used, substantial variation in live birth rates by patient (e.g., women aged < or =40 years) and treatment characteristics (e.g., ovulatory dysfunction, endometriosis, or unexplained infertility) was observed. The risk for a multiple-birth delivery was highest for women who underwent ART transfer procedures using freshly fertilized embryos from either donor eggs (42%) or from their own eggs (36%). Among ART transfer procedures in which the patient's own eggs were used, an inverse relation existed between multiple-birth risk and patient age. Number of embryos transferred and embryo availability (an indicator of embryo quality) were also strong predictors of multiple-birth risk. Of the 40,687 infants born, 46% were twins, and 8% were triplet and higher order multiples. The total multiple-infant birth rate was 53%. Approximately 1% of U.S. infants born in 2001 were conceived through ART. Those infants accounted for 16% of multiple births nationally. INTERPRETATION: Whether an ART procedure resulted in a pregnancy and live-birth delivery varied according to different patient and treatment factors. ART poses a major risk for multiple births. This risk varied according to the patient's age, the type of ART procedure performed, the number of embryos transferred, and embryo availability (an indicator of embryo quality). PUBLIC HEALTH ACTION: ART-related multiple births represent a sizable proportion of all multiple births nationally and in selected states. Efforts should be made to limit the number of embryos transferred for patients undergoing ART. PMID- 15123983 TI - Malaria surveillance--United States, 2002. AB - PROBLEM/CONDITION: Malaria is caused by any of four species of intraerythrocytic protozoa of the genus Plasmodium (i.e., P. falciparum, P. vivax, P. ovale, or P. malariae). These parasites are transmitted by the bite of an infective female Anopheles species mosquito. The majority of malaria infections in the United States occur among persons who have traveled to areas with ongoing transmission. In the United States, cases can occur through exposure to infected blood products, by congenital transmission, or by local mosquitoborne transmission. Malaria surveillance is conducted to identify episodes of local transmission and to guide prevention recommendations for travelers. PERIOD COVERED: This report covers cases with onset of illness in 2002. DESCRIPTION OF SYSTEM: Malaria cases confirmed by blood film are reported to local and state health departments by health-care providers or laboratory staff. Case investigations are conducted by local and state health departments, and reports are transmitted to CDC through the National Malaria Surveillance System (NMSS). Data from NMSS serve as the basis for this report. RESULTS: CDC received reports of 1,337 cases of malaria with an onset of symptoms in 2002 among persons in the United States or one of its territories. This number represents a decrease of 3.3% from the 1,383 cases reported for 2001. P. falciparum, P. vivax, P. malariae, and P. ovale were identified in 52.3%, 25.4%, 2.8%, and 2.8% of cases, respectively. Eleven patients (0.8% of total) were infected by > or =2 species. The infecting species was unreported or undetermined in 213 (15.9%) cases. Compared with 2001, the number of reported malaria cases acquired in Asia (n = 171) and Africa (n = 903) increased by 4.3% and 1.9%, respectively, whereas the number of cases acquired in the Americas (n = 141) decreased by 41.2%. Of 849 U.S. civilians who acquired malaria abroad, 317 (37.3%) reported that they had followed a chemoprophylactic drug regimen recommended by CDC for the area to which they had traveled. Five patients became infected in the United States, one through congenital transmission, one probable transfusion-related, and three whose infection cannot be linked epidemiologically to secondary cases. Eight deaths were attributed to malaria. All deaths were caused by P. falciparum. INTERPRETATION: The 3.3% decrease in malaria cases in 2002, compared with 2001, resulted primarily from a marked decrease in cases acquired in the Americas, but this decrease was offset somewhat by an increase in the number of cases acquired in Africa and Asia. This limited decrease probably represents year-to-year variation in malaria cases, but also could have resulted from local changes in disease transmission, decreased travel to malaria-endemic regions, fluctuation in reporting to state and local health departments, or an increased use of effective antimalarial chemoprophylaxis. In the majority of reported cases, U.S. civilians who acquired infection abroad were not on an appropriate chemoprophylaxis regimen for the country in which they acquired malaria. PUBLIC HEALTH ACTION: Additional information was obtained concerning the eight fatal cases and the five infections acquired in the United States. Persons traveling to a malarious area should take one of the recommended chemoprophylaxis regimens appropriate for the region of travel, and travelers should use personal protection measures to prevent mosquito bites. Any person who has been to a malarious area and who subsequently experiences a fever or influenza-like symptoms should seek medical care immediately and report their travel history to the clinician; investigation should include a blood-film test for malaria. Malaria infections can be fatal if not diagnosed and treated promptly. Recommendations concerning malaria prevention can be obtained from CDC by calling the Malaria Hotline at 770-488-7788 or by accessing CDC's Internet site at http://www.cdc.gov/travel. PMID- 15123984 TI - Diagnosis and management of foodborne illnesses: a primer for physicians and other health care professionals. PMID- 15123985 TI - Increases in fluoroquinolone-resistant Neisseria gonorrhoeae among men who have sex with men--United States, 2003, and revised recommendations for gonorrhea treatment, 2004. AB - In the United States, an estimated 700,000-800,000 persons are infected with Neisseria gonorrhoeae each year. Since 1993, CDC has recommended use of fluoroquinolones (i.e., ciprofloxacin, floxacin, or levofloxacin) for gonorrhea treatment. Fluoroquinolone therapy is used frequently because it is an inexpensive, oral, and single-dose therapy. However, because of increased prevalence of fluoroquinolone-resistant N. gonorrhoeae (QRNG) in Asia, the Pacific Islands (including Hawaii), and California, fluoroquinolones are no longer recommended for treating gonorrhea acquired in those locations. This report describes increases in QRNG among men who have sex with men (MSM) in Massachusetts, New York City, and 30 sites surveyed by the Gonococcal Isolate Surveillance Project (GISP) during 2003. CDC recommends that clinicians no longer use fluoroquinolones as a first-line treatment for gonorrhea in MSM. PMID- 15123986 TI - Preliminary FoodNet data on the incidence of infection with pathogens transmitted commonly through food--selected sites, United States, 2003. AB - In the United States, an estimated 76 million persons contract foodborne and other acute diarrheal illnesses each year. CDC's Emerging Infections Program Foodborne Diseases Active Surveillance Network (FoodNet) collects data on diseases caused by enteric pathogens transmitted commonly through food in nine U.S. sites. FoodNet quantifies and monitors the incidence of these infections by conducting active surveillance for laboratory-diagnosed illness. This report describes preliminary surveillance data for 2003 and compares them with 1996-2002 data. The data indicate substantial declines in the incidence of infections caused by Campylobacter, Cryptosporidium parvum, Escherichia coli O157, Salmonella, and Yersinia enterocolitica. These data represent progress toward meeting the 2010 national health objectives of reducing the incidence of foodborne infections (objective nos. 10.1a, 10.1b, and 10.1d). However, increased efforts are needed to reduce further the incidence of foodborne illnesses, particularly among children. PMID- 15123987 TI - Progress toward poliomyelitis eradication--Nigeria, January 2003-March 2004. AB - Since the 1988 World Health Assembly resolution to eradicate poliomyelitis, three World Health Organization (WHO) regions (Americas, Western Pacific, and European) have been certified polio-free, and the number of countries where polio is endemic has decreased from 125 in 1988 to six in 2003 (Afghanistan, Egypt, India, Niger, Nigeria, and Pakistan). In 2003, Nigeria, the most populous country of the African continent (2003 population projected from 1991 census report: 125 million), reported 355 wild poliovirus (WPV) cases, accounting for 45% of cases reported globally and >80% of cases reported from the African Region (AFR). This report summarizes progress toward polio eradication in Nigeria during January 2003-March 2004. The findings indicate the urgent need to implement high-quality supplementary immunization activities (SIAs) in Nigeria to interrupt ongoing WPV transmission. PMID- 15123988 TI - Summary of notifiable diseases--United States, 2002. PMID- 15123989 TI - Bioinformatic approaches for identification and characterization of olfactomedin related genes with a potential role in pathogenesis of ocular disorders. AB - PURPOSE: To identify olfactomedin domain containing proteins, which are expressed in the eye and have similarity to myocilin, to test as potential candidates for eye diseases. Most of the mutations in myocilin causing primary open angle glaucoma are located in the olfactomedin domain. In vitro experiments demonstrated interaction between optimedin and myocilin through the conserved olfactomedin domains of the proteins in rats, and it was speculated that optimedin might have a role in the pathogenesis of ocular disorders. Hence, we aimed to identify myocilin related human proteins having conserved olfactomedin domains with potential to interact between them and examine the expression patterns in the eye by bioinformatics approaches. This endeavor would have the potential to identify new candidate genes for eye diseases in general and glaucoma in particular to be tested by wet-lab experiments. METHODS: Proteins with homology to myocilin were selected by BLASTp at the NCBI server. cDNA sequences and corresponding genomic contigs were retrieved. Pairwise BLAST was done to investigate the gene structure. The human EST database and NEIBank were searched against the selected cDNAs to look for tissue specific expression of the transcripts. RESULTS: The study led to the identification of three groups of proteins encoded by three different genes; Noelin 1 (9q34.3), Noelin 2 (19p13.2), and Noelin 3 (1p22) encompassing 45,575 bp, 82,679 bp, and 1,93,421 bp of the genomic sequence, respectively. Genomic structures, alternate usage of exons, and molecular evolution of the Noelins were determined. Similar structures of the genes, splicing patterns and high levels of homology shed light on the relatedness and molecular evolution of this group of olfactomedin related proteins. Strikingly, however, Noelin 1 and Noelin 3 were found to be expressed as multiple splice variants while only a single spliced transcript could be identified for Noelin 2. A human EST database search suggested the expression of all three Noelin genes in the brain but only two (Noelin 1 and Noelin 2) in the eye despite experimental evidence for expression of Noelin 3 in ocular tissue. Myocilin was determined to have similar levels (60-61%) of homology with all three Noelin gene products (Noelin 1_v1, Noelin 2_v1, and Noelin 3_v1) at the conserved olfactomedin domains. CONCLUSIONS: Mammalian Noelin 1 evolved from its precursor, followed by evolution of Noelin 3 and Noelin 2 by gene duplication events. Myocilin might have evolved from Noelin 2 by gene duplication followed by exon fusion. Noelin 1 and Noelin 2 could be tested as candidate genes for eye diseases based on their expressions in the eye and shared olfactomedin domains with Myocilin in the C-termini of the respective proteins. PMID- 15123990 TI - Functional analyses of mutant recessive GUCY2D alleles identified in Leber congenital amaurosis patients: protein domain comparisons and dominant negative effects. AB - PURPOSE: Recessive mutations in GUCY2D, the gene encoding the retinal guanylyl cyclase protein, RetGC-1, have been shown to cause Leber Congenital Amaurosis (LCA), a severe retinal dystrophy. The purpose of this study was to determine the functional consequences of selected mutations in GUCY2Dlinked to LCA. The mutations investigated in this study map to the catalytic domain (P858S, L954P) and the extracellular domain (C105Y, L325P) of RetGC-1. METHODS: All four mutations were introduced into the in vitro expression plasmid, pRC-CMV human RetGC-1, and expressed in HEK-293 cells. We assayed the abilities of the mutant cyclases to generate cGMP (basal activity), and to be activated by guanylyl cyclase activating proteins (GCAP-1 and GCAP-2). Additionally, we co-expressed the catalytic domain mutations (P858S and L954P) with a wild-type allele to test for dominant negative effects on wild-type RetGC-1. RESULTS: The P858S and L954P mutations, both in highly conserved residues of the catalytic domain of RetGC-1, severely impair basal, GCAP-1, and GCAP-2 stimulated catalytic activity of the enzyme. In addition, when co-expressed with the wild-type allele, both catalytic domain mutations act as dominant negative proteins and reduce the activity of wild-type RetGC-1. The basal activities of the C105Y and L325P mutants are unaltered, but GCAP-1 and GCAP-2 stimulated cyclase activities are reduced approximately 50%. CONCLUSIONS: GUCY2D mutations from LCA patients have distinct functional consequences on RetGC-1 catalytic activity in vitro. Our analyses showed that the catalytic domain mutations cause a marked reduction in cyclase activity, while the extracellular domain mutations moderately reduce activity. The catalytic domain mutant alleles cause dominant negative effects, indicating that the functionality of RetGC-1 is compromised even in heterozygotes. This is consistent with abnormalities in cone electroretinograms (ERGs) detected in obligate heterozygous GUCY2D parents that carry the L954P mutation. PMID- 15123994 TI - Hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy is a fascinating disease of marked heterogeneity. Hypertrophic cardiomyopathy was originally characterized by massive myocardial hypertrophy in the absence of known cause, a dynamic left ventricular outflow obstruction, and increased risk of sudden death. It is now well accepted that multiple mutations in genes encoding for the cardiac sarcomere are responsible for the disease. Complex morphologic and pathophysiological differences, disparate natural history studies, and novel treatment strategies underscore the challenge to the practicing cardiologist when faced with the management of the patient with hypertrophic cardiomyopathy. PMID- 15123996 TI - Pharmacogenomics in drug development: societal and technical aspects. PMID- 15123997 TI - Lumbar spinal angiolipomas: report of two cases and review of the literature. AB - STUDY DESIGN: Case report and review of the literature. OBJECTIVES: To describe two patients with angiolipoma in the ventral aspect of the lumbar epidural space, to discuss the clinical, radiologic, and surgical features of these lesions, and to review previously reported cases. SETTING: Rome, Italy. METHODS: Two cases, a 60-year-old man and a 54-year-old woman presented with lumbar-sciatic pain but with no abnormal neurological signs. Investigation (CT and MRI) demonstrated lumbar tumours. RESULTS: Laminectomy and excision of the tumors were performed, and symptoms improved immediately. CONCLUSIONS: Magnetic resonance imaging with suppression fat sequences allows the recognition of these lesions. The prognosis after surgical removal of spinal angiolipoma is favorable. PMID- 15123998 TI - Dantrolene and pleural effusion: case report and review of literature. AB - STUDY DESIGN: Single-subject case (a quadriplegic female, 56 years). OBJECTIVES: To describe a new case of eosinophilic pleural effusion induced by dantrolene chronic administration. SETTING: Physical medicine and rehabilitation unit in a teaching hospital, France. METHODS: Diagnosis of an eosinophilic pleural effusion induced by dantrolene without any respiratory symptoms, except a decrease of breath sounds on the right lung base. RESULTS: Chest radiograph revealed a right sided pleural effusion, and blood cell count a significant peripheral eosinophilia. Thoracenthesis contained 85% of eosinophils. The other explorations eliminated other causes of pleural effusion. The diagnosis of drug-induced effusion was almost sure and led us to discontinue the dantrolene. After 3 months, she had completely recovered. These characteristics, similar to the eight other cases described in the literature, are essential for the diagnosis of pleural effusion induced by dantrolene. CONCLUSION: Dantrolene, a long-acting skeletal muscle relaxant, is well known to induce liver side effects but it can also induce pleural pericarditis. The pathogenesis is still not clearly identified, but similarities of chemical structures of dantrolene and nitrofurantoine make us think that it could be the same mechanism. The association between dantrolene and nitrofurantoine may have contributed to the expression of the pleural effusion. PMID- 15123999 TI - Brucellar spondylitis: a rare cause of spinal cord compression. AB - STUDY DESIGN: A case report. OBJECTIVES: To present and discuss some of the difficulties in the diagnosis of brucellar spondylitis. SETTING: Ankara University, Ibni Sina Hospital, Turkey. METHODS: We report a patient with paraplegia, misdiagnosed as having a malignancy or tuberculosis who actually suffered from brucellar spondylitis. Diagnosis was established by her history and a compatible clinical picture together with a standard tube agglutination (Wright test) titer of > or =1/160 of antibodies for brucellosis. The patient was treated with oral doxycycline, rifampicin, and ciprofloxacin combination. RESULTS: At the end of the treatment, the blood Brucella Wright and anti-human globulin T titer levels decreased. Her lower limb weakness improved. She could walk, and climb stairs with the help of a cane. Urinary retention and fecal incontinence also resolved. CONCLUSION: Brucellosis is a systemic infection involving the musculoskeletal and nervous systems. Spondylitis frequently occurs in elderly patients. An early diagnosis of brucellar spondylitis can often be difficult. In endemic regions, as in the case of our country, brucellar spondylitis should always be considered in the differential diagnosis of older patients with back pain and constitutional symptoms. An early diagnosis will help to prevent the development of more severe complications such as spinal cord compression. PMID- 15124000 TI - Stepping-like movements in humans with complete spinal cord injury induced by epidural stimulation of the lumbar cord: electromyographic study of compound muscle action potentials. AB - STUDY DESIGN: It has been previously demonstrated that sustained nonpatterned electric stimulation of the posterior lumbar spinal cord from the epidural space can induce stepping-like movements in subjects with chronic, complete spinal cord injury. In the present paper, we explore physiologically related components of electromyographic (EMG) recordings during the induced stepping-like activity. OBJECTIVES: To examine mechanisms underlying the stepping-like movements activated by electrical epidural stimulation of posterior lumbar cord structures. MATERIALS AND METHODS: The study is based on the assessment of epidural stimulation to control spasticity by simultaneous recordings of the electromyographic activity of quadriceps, hamstrings, tibialis anterior, and triceps surae. We examined induced muscle responses to stimulation frequencies of 2.2-50 Hz in 10 subjects classified as having a motor complete spinal cord injury (ASIA A and B). We evaluated stimulus-triggered time windows 50 ms in length from the original EMG traces. Stimulus-evoked compound muscle action potentials (CMAPs) were analyzed with reference to latency, amplitude, and shape. RESULTS: Epidural stimulation of the posterior lumbosacral cord recruited lower limb muscles in a segmental-selective way, which was characteristic for posterior root stimulation. A 2.2 Hz stimulation elicited stimulus-coupled CMAPs of short latency which were approximately half that of phasic stretch reflex latencies for the respective muscle groups. EMG amplitudes were stimulus-strength dependent. Stimulation at 5-15 and 25-50 Hz elicited sustained tonic and rhythmic activity, respectively, and initiated lower limb extension or stepping-like movements representing different levels of muscle synergies. All EMG responses, even during burst-style phases were composed of separate stimulus-triggered CMAPs with characteristic amplitude modulations. During burst-style phases, a significant increase of CMAP latencies by about 10 ms was observed. CONCLUSION: The muscle activity evoked by epidural lumbar cord stimulation as described in the present study was initiated within the posterior roots. These posterior roots muscle reflex responses (PRMRRs) to 2.2 Hz stimulation were routed through monosynaptic pathways. Sustained stimulation at 5-50 Hz engaged central spinal PRMRR components. We propose that repeated volleys delivered to the lumbar cord via the posterior roots can effectively modify the central state of spinal circuits by temporarily combining them into functional units generating integrated motor behavior of sustained extension and rhythmic flexion/extension movements. This study opens the possibility for developing neuroprostheses for activation of inherent spinal networks involved in generating functional synergistic movements using a single electrode implanted in a localized and stable region. PMID- 15124001 TI - Potential impact of orthotic gait exercise on natural killer cell activities in thoracic level of spinal cord-injured patients. AB - STUDY DESIGN: Prospective before-after trial. OBJECTIVE: To examine the changes of natural killer (NK) cell activity in response to orthotic gait exercise in thoracic level of spinal cord-injured (SCI) patients. SETTING: National Rehabilitation Center for Persons with Disabilities, Japan. METHODS: In all, 10 thoracic level of SCI patients (ranging Th5-Th12), who experienced orthotic gait training, participated in this study. NK cell activity at an effector:target (E/T) ratio (20:1) was examined in a sample of peripheral blood taken before and just after orthotic gait exercise for 20 min. On a separate day, to evaluate the physical intensity of the orthotic gait exercise, cardiorespiratory responses at rest and during exercise were measured. RESULTS: The resting value of the NK cell activity in our SCI patients was remarkably lower than that in normal subjects reported in previous studies. The NK cell activity was significantly increased through a 20 min orthotic gait exercise (pre versus post; 12.7+/-5.28 versus 17.76+/-6.71, P<0.05). The steady-state value of oxygen (VO2) and heart rate (HR) were 18.13+/-3.92 ml/kg and 142.53+/-19.84 b/min, respectively. It was noteworthy that a patient who showed decrement of NK cell activity in response to exercise had the highest level of injury (Th5), and showed the higher energy cost of orthotic gait. CONCLUSION: These findings suggested that the orthotic gait exercise has the potential to enhance the immune function for SCI persons, although patients with a higher level of SCI may have some difficulties. SPONSORSHIP: Mitsui Sumitomo Insurance Welfare Foundation PMID- 15124002 TI - Importance of specialised spinal units. PMID- 15124003 TI - Androgen signaling and post-transcriptional downregulation of Bcl-2 in androgen unresponsive prostate cancer. AB - We previously characterized the LNCaP prostate cancer progression model and showed that despite loss of Bcl-2 protein in the androgen-unresponsive LNCaP unresponsive (UR) cells, these cells maintained an increased resistance to the induction of apoptosis. Since the loss of Bcl-2 protein coincided with the progression to androgen-unresponsiveness, we sought to determine if Bcl-2 expression was regulated through androgen signaling pathways. LNCaP-responsive (R) and -UR cells grown in charcoal-stripped serum conditions for 3 months differentiated to a neuroendocrine (NE)-like morphology. Under these conditions, LNCaP-UR cells regained Bcl-2 protein expression, and LNCaP-R cells overexpressed Bcl-2. Chronic exposure to casodex resulted in differentiation of both LNCaP-R and -UR cells to the NE-type morphology accompanied by a marked downregulation of Bcl-2 protein, while Bax protein levels were unchanged. Downregulation of Bcl-2 was post-transcriptional since Bcl-2 message levels were unchanged in LNCaP cells treated with casodex. These data suggest that Bcl-2 is post-transcriptionally modulated by androgen signaling pathways in LNCaP cells. PMID- 15124004 TI - Variants in the catechol-o-methyltransferase (COMT) gene are associated with schizophrenia in Irish high-density families. AB - The enzyme catechol-o-methyltransferase (COMT) transfers a methyl group from adenosylmethionine to catecholamines including the neurotransmitters dopamine, epinephrine and norepinephrine. This methylation results in the degradation of catecholamines. The involvement of the COMT gene in the metabolic pathway of these neurotransmitters has made it an attractive candidate gene for many psychiatric disorders. In this article, we reported our study of association of COMT with schizophrenia in Irish families with a high density of schizophrenia. Three single nucleotide polymorphisms (SNPs) were genotyped for the 274 such families and within-family transmission disequilibrium tests were performed. SNP rs4680, which is the functional Val/Met polymorphism, showed modest association with the disease by the TRANSMIT, FBAT and PDT programs, while the other two SNPs were negative. These SNPs showed lower level of LDs with each other in the Irish subjects than in Ashkenazi Jews. Haplotype analysis indicated that a haplotype, haplotype A-G-A for SNPs rs737865-rs4680-rs165599, was preferentially transmitted to the affected subjects. This was different from the reported G-G-G haplotype found in Ashkenazi Jews, but both haplotypes shared the Val allele. We concluded that COMT gene is associated with schizophrenia and carries a small but significant risk to the susceptibility in the Irish subjects. PMID- 15124006 TI - SNP and haplotype analysis of a novel tryptophan hydroxylase isoform (TPH2) gene provide evidence for association with major depression. AB - Tryptophan hydroxylase (TPH), being the rate-limiting enzyme in the biosynthesis of serotonin plays a major role as candidate gene in several psychiatric disorders. Recently, a second TPH isoform (TPH2) was identified in mice, which was exclusively present in the brain. In a previous post-mortem study of our own group, we could demonstrate that TPH2 is also expressed in the human brain, but not in peripheral tissues. This is the first report of an association study between polymorphisms in the TPH2 gene and major depression (MD). We performed single-nucleotide polymorphism (SNP), haplotype and linkage disequlibrium studies on 300 depressed patients and 265 healthy controls with 10 SNPs in the TPH2 gene. Significant association was detected between one SNP (P=0.0012, global P=0.0051) and MD. Haplotype analysis produced additional support for association (P<0.0001, global P=0.0001). Our findings provide evidence for an involvement of genetic variants of the TPH2 gene in the pathogenesis of MD and might be a hint on the repeatedly discussed duality of the serotonergic system. These results may open up new research strategies for the analysis of the observed disturbances in the serotonergic system in patients suffering from several other psychiatric disorders. PMID- 15124007 TI - What can laboratory notifications tell us about Chlamydia infection? PMID- 15124008 TI - Capacity building infrastructure grants workshop. PMID- 15124009 TI - Meningococcal disease in New South Wales, 1991-2002. PMID- 15124010 TI - Communicable diseases report, New South Wales, for December 2003 and January 2004. PMID- 15124011 TI - Hard-hearted CRP. PMID- 15124012 TI - Lost in transdifferentiation. AB - What are the true origins of the smooth muscle cells (SMCs) present in the intimal lesions of transplant arteriosclerosis? A new study in the JCI shows that Sca-1(+) cells purified from the mouse aortic root can migrate through an irradiated vein graft to the neointima of the vessel and transdifferentiate to express the early SMC differentiation marker gene SM22. Do Sca-1(+) cells transdifferentiate into SMC-like cells, or is activation of SMC marker genes a consequence of fusion of these cells with preexisting SMCs, a possibility raised by results of studies of adult stem cells in animal models of liver regeneration ? Or could this be bona fide transdifferentiation that recapitulates the pathologic processes in humans? PMID- 15124013 TI - Anemia of inflammation: the cytokine-hepcidin link. AB - The anemia of inflammation, commonly observed in patients with chronic infections, malignancy, trauma, and inflammatory disorders, is a well-known clinical entity. Until recently, we understood little about its pathogenesis. It now appears that the inflammatory cytokine IL-6 induces production of hepcidin, an iron-regulatory hormone that may be responsible for most or all of the features of this disorder. PMID- 15124014 TI - Hold the antioxidants and improve plasma lipids? AB - Intrahepatic proteolysis is a major determinant of secretion of ApoB-containing lipoproteins into plasma. Stimulation of post-ER presecretory proteolysis (PERPP) of ApoB by n-3 polyunsaturated fatty acids has been found to result in reduced secretion of VLDL particles by hepatocytes. A new study has shown that this stimulation is promoted by pro-oxidant conditions that result in increased hepatic lipid hydroperoxide content. Conversely, PERPP is suppressed by antioxidants and by saturated fatty acids, which are not susceptible to lipid peroxidation. Hence reduction of oxidative stress may have the unexpected side effect of increasing plasma lipid levels. PMID- 15124015 TI - Dysbindin-1 and schizophrenia: from genetics to neuropathology. AB - The gene encoding dysbindin-1 has recently been implicated in susceptibility to schizophrenia. In this issue of the JCI, Talbot et al. show that, contrary to expectations, dysbindin-1 is located presynaptically in glutamatergic neurons and is reduced at these locations in schizophrenia. Further studies of dysbindin-1 and the proteins with which it interacts can be expected to throw light on the pathogenesis of schizophrenia. PMID- 15124016 TI - Abundant progenitor cells in the adventitia contribute to atherosclerosis of vein grafts in ApoE-deficient mice. AB - Recent evidence indicates that vascular progenitor cells may be the source of smooth muscle cells (SMCs) that accumulate in atherosclerotic lesions, but the origin of these progenitor cells is unknown. To explore the possibility of vascular progenitor cells existing in adults, a variety of tissues from ApoE deficient mice were extensively examined. Immunohistochemical staining revealed that the adventitia in aortic roots harbored large numbers of cells having stem cell markers, e.g., Sca-1(+) (21%), c-kit(+) (9%), CD34(+) (15%), and Flk1(+) cells (4%), but not SSEA-1(+) embryonic stem cells. Explanted cultures of adventitial tissues using stem cell medium displayed a heterogeneous outgrowth, for example, islands of round-shaped cells surrounded by fibroblast-like cell monolayers. Isolated Sca-1(+) cells were able to differentiate into SMCs in response to PDGF-BB stimulation in vitro. When Sca-1(+) cells carrying the LacZ gene were transferred to the adventitial side of vein grafts in ApoE-deficient mice, beta-gal(+) cells were found in atherosclerotic lesions of the intima, and these cells enhanced the development of the lesions. Thus, a large population of vascular progenitor cells existing in the adventitia can differentiate into SMCs that contribute to atherosclerosis. Our findings indicate that ex vivo expansion of these progenitor cells may have implications for cellular, genetic, and tissue engineering approaches to vascular disease. PMID- 15124017 TI - Hematopoietic myelomonocytic cells are the major source of hepatocyte fusion partners. AB - Several recent reports have demonstrated that transplantation of bone marrow cells can result in the generation of functional hepatocytes. Cellular fusion between bone marrow-derived cells and host hepatocytes has been shown to be the mechanism of this phenomenon. However, the exact identity of the bone marrow cells that mediate cellular fusion has remained undetermined. Here we demonstrate that the hematopoietic progeny of a single hematopoietic stem cell (HSC) is sufficient to produce functional hepatic repopulation. Furthermore, transplantation of lymphocyte-deficient bone marrow cells and in vivo fate mapping of the myeloid lineage revealed that HSC-derived hepatocytes are primarily derived from mature myelomonocytic cells. In addition, using a Cre/lox based strategy, we directly demonstrate that myeloid cells spontaneously fuse with host hepatocytes. Our findings raise the possibility that differentiated myeloid cells may be useful for future therapeutic applications of in vivo cellular fusion. PMID- 15124018 TI - IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin. AB - Hypoferremia is a common response to systemic infections or generalized inflammatory disorders. In mouse models, the development of hypoferremia during inflammation requires hepcidin, an iron regulatory peptide hormone produced in the liver, but the inflammatory signals that regulate hepcidin are largely unknown. Our studies in human liver cell cultures, mice, and human volunteers indicate that IL-6 is the necessary and sufficient cytokine for the induction of hepcidin during inflammation and that the IL-6-hepcidin axis is responsible for the hypoferremia of inflammation. PMID- 15124019 TI - Lipid peroxidation and oxidant stress regulate hepatic apolipoprotein B degradation and VLDL production. AB - How omega-3 and omega-6 polyunsaturated fatty acids (PUFAs) lower plasma lipid levels is incompletely understood. We previously showed that marine omega-3 PUFAs (docosahexaenoic acid [DHA] and eicosapentaenoic acid) stimulate a novel pathway, post-ER presecretory proteolysis (PERPP), that degrades apolipoprotein B100 (ApoB100), thereby reducing lipoprotein secretion from liver cells. To identify signals stimulating PERPP, we examined known actions of omega-3 PUFA. In rat hepatoma or primary rodent hepatocytes incubated with omega-3 PUFA, cotreatment with the iron chelator desferrioxamine, an inhibitor of iron-dependent lipid peroxidation, or vitamin E, a lipid antioxidant, suppressed increases in thiobarbituric acid-reactive substances (TBARSs; a measure of lipid peroxidation products) and restored ApoB100 recovery and VLDL secretion. Moreover, omega-6 and nonmarine omega-3 PUFA, also prone to peroxidation, increased ApoB100 degradation via intracellular induction of TBARSs. Even without added fatty acids, degradation of ApoB100 in primary hepatocytes was blocked by desferrioxamine or antioxidant cotreatment. To extend these results in vivo, mice were infused with DHA, which increased hepatic TBARSs and reduced VLDL-ApoB100 secretion. These results establish a novel link between lipid peroxidation and oxidant stress with ApoB100 degradation via PERPP, and may be relevant to the hypolipidemic actions of dietary PUFAs, the basal regulation of ApoB100 secretion, and hyperlipidemias arising from ApoB100 overproduction. PMID- 15124020 TI - Impaired pancreatic growth, beta cell mass, and beta cell function in E2F1 (-/- )mice. AB - We evaluated the effects of E2F1 on glucose homeostasis using E2F1(-/-) mice. E2F1(-/-) mice show an overall reduction in pancreatic size as the result of impaired postnatal pancreatic growth. Furthermore, these animals have dysfunctional beta cells, linked to impaired PDX-1 activity. Because of the disproportionate small pancreas and dysfunctional islets, E2F1(-/-) mice secrete insufficient amounts of insulin in response to a glucose load, resulting in glucose intolerance. Despite this glucose intolerance, E2F1(-/-) mice do not develop overt diabetes mellitus because they have insulin hypersensitivity, which is secondary to a diminished adipose tissue mass and altered adipocytokine levels, which compensates for the defect in insulin secretion. These data demonstrate that factors controlling cell proliferation, such as E2F1, determine pancreatic growth and function, subsequently affecting metabolic homeostasis. PMID- 15124021 TI - Bacterial flagellin is a dominant antigen in Crohn disease. AB - Chronic intestinal inflammation, as seen in inflammatory bowel disease (IBD), results from an aberrant and poorly understood mucosal immune response to the microbiota of the gastrointestinal tract in genetically susceptible individuals. Here we used serological expression cloning to identify commensal bacterial proteins that could contribute to the pathogenesis of IBD. The dominant antigens identified were flagellins, molecules known to activate innate immunity via Toll like receptor 5 (TLR5), and critical targets of the acquired immune system in host defense. Multiple strains of colitic mice had elevated serum anti-flagellin IgG2a responses and Th1 T cell responses to flagellin. In addition, flagellin specific CD4(+) T cells induced severe colitis when adoptively transferred into naive SCID mice. Serum IgG to these flagellins, but not to the dissimilar Salmonella muenchen flagellin, was elevated in patients with Crohn disease, but not in patients with ulcerative colitis or in controls. These results identify flagellins as a class of immunodominant antigens that stimulate pathogenic intestinal immune reactions in genetically diverse hosts and suggest new avenues for the diagnosis and antigen-directed therapy of patients with IBD. PMID- 15124022 TI - Prevention of gastrointestinal tumors based on adenomatous polyposis coli gene mutation by dendritic cell vaccine. AB - Here we describe the effect of immunization with dendritic cells loaded with syngeneic tumor cells (DC/Ts) by polyethylene glycol treatment, on tumor development in adenomatous polyposis coli (APC) gene mutant mouse models, APC1309 and APC(Min-/+), in which adenomatous polyps of the gastrointestinal tracts develop with a high incidence. Treatment with DC/Ts prevented the development of gastrointestinal tumors, and coadministration of DC/Ts and IL-12 caused a further reduction in tumor incidence. Splenocytes from APC1309 mice treated with DC/Ts and IL-12 showed no cytotoxic activity toward the tumor cells, but serum antibody specific to them was detected. IgG from the treated mice exhibited cytotoxic activity against the tumor cells in vitro. Predominance of Th2 cell response over Th1 response was also suggested by ELISPOT assays in the treated mice. Depletion in vivo of CD4(+) T cells, not CD8(+) T cells, by the intraperitoneal administration of corresponding mAb's decreased the antitumor effect of DC/T inoculation. Immunofluorescence microscopic studies showed that Ig was attached to tumor cells in mice treated with DC/Ts and IL-12. These findings indicate that DC/T vaccination prevents tumor development through APC gene mutation and that its preventive effects are mediated by humoral antitumor immunity. PMID- 15124023 TI - Pre-B cell colony-enhancing factor inhibits neutrophil apoptosis in experimental inflammation and clinical sepsis. AB - Pre-B cell colony-enhancing factor (PBEF) is a highly conserved 52-kDa protein, originally identified as a growth factor for early stage B cells. We show here that PBEF is also upregulated in neutrophils by IL-1beta and functions as a novel inhibitor of apoptosis in response to a variety of inflammatory stimuli. Induction of PBEF occurs 5-10 hours after LPS exposure. Prevention of PBEF translation with an antisense oligonucleotide completely abrogates the inhibitory effects of LPS, IL-1, GM-CSF, IL-8, and TNF-alpha on neutrophil apoptosis. Immunoreactive PBEF is detectable in culture supernatants from LPS-stimulated neutrophils, and a recombinant PBEF fusion protein inhibits neutrophil apoptosis. PBEF is also expressed in neutrophils from critically ill patients with sepsis in whom rates of apoptosis are profoundly delayed. Expression occurs at higher levels than those seen in experimental inflammation, and a PBEF antisense oligonucleotide significantly restores the normal kinetics of apoptosis in septic polymorphonuclear neutrophils. Inhibition of apoptosis by PBEF is associated with reduced activity of caspases-8 and -3, but not caspase-9. These data identify PBEF as a novel inflammatory cytokine that plays a requisite role in the delayed neutrophil apoptosis of clinical and experimental sepsis. PMID- 15124024 TI - The MHC class I-like Fc receptor promotes humorally mediated autoimmune disease. AB - The MHC class I family-like Fc receptor, FcRn, is normally responsible for extending the life span of serum IgG Ab's, but whether this molecule contributes to autoimmune pathogenesis remains speculative. To determine directly whether this function contributes to humoral autoimmune disease, we examined whether a deficiency in the FcRn heavy chain influences autoimmune arthritis in the K/BxN mouse model. FcRn deficiency conferred either partial or complete protection in the arthritogenic serum transfer and the more aggressive genetically determined K/BxN autoimmune arthritis models. The protective effects of an FcRn deficiency could be overridden with excessive amounts of pathogenic IgG Ab's. The therapeutic saturation of FcRn by high-dose intravenous IgG (IVIg) also ameliorated arthritis, directly implicating FcRn blockade as a significant mechanism of IVIg's anti-inflammatory action. The results suggest that FcRn is a potential therapeutic target that links the initiation and effector phases of humoral autoimmune disease. PMID- 15124025 TI - The cyclin-dependent kinase inhibitor p57(Kip2) mediates proliferative actions of PTHrP in chondrocytes. AB - Parathyroid hormone-related peptide (PTHrP) is a positive regulator of chondrocyte proliferation during bone development. In embryonic mice lacking PTHrP, chondrocytes stop proliferating prematurely, with accelerated differentiation. Because the bone phenotype of mice lacking the cyclin-dependent kinase inhibitor p57(Kip2) is the opposite of the PTHrP-null phenotype, we hypothesized that PTHrP's proliferative actions in chondrocytes might be mediated by opposing p57. We generated p57/PTHrP-null embryos, which showed partial rescue of the PTHrP-null phenotype. There was reversal of the loss of proliferative chondrocytes in most bones, with reversal of the accelerated differentiation that occurs in the PTHrP-null phenotype. p57 mRNA and protein were upregulated in proliferative chondrocytes in the absence of PTHrP. Metatarsal culture studies confirmed the action of PTHrP to decrease p57 mRNA and protein levels in a model in which parathyroid hormone (PTH), used as an analog of PTHrP, increased chondrocyte proliferation rate and the length of the proliferative domain. PTH treatment of p57-null metatarsals had no effect on proliferation rate in round proliferative chondrocytes but still stimulated proliferation in columnar chondrocytes. These studies suggest that the effects of PTHrP on both the rate and extent of chondrocyte proliferation are mediated, at least in part, through suppression of p57 expression. PMID- 15124026 TI - Deletion of vanilloid receptor 1-expressing primary afferent neurons for pain control. AB - Control of cancer, neuropathic, and postoperative pain is frequently inadequate or compromised by debilitating side effects. Inhibition or removal of certain nociceptive neurons, while retaining all other sensory modalities and motor function, would represent a new therapeutic approach to control severe pain. The enriched expression of transient receptor potential cation channel, subfamily V, member 1 (TRPV1; also known as the vanilloid receptor, VR1) in nociceptive neurons of the dorsal root and trigeminal ganglia allowed us to test this concept. Administration of the potent TRPV1 agonist resiniferatoxin (RTX) to neuronal perikarya induces calcium cytotoxicity by opening the TRPV1 ion channel and selectively ablates nociceptive neurons. This treatment blocks experimental inflammatory hyperalgesia and neurogenic inflammation in rats and naturally occurring cancer and debilitating arthritic pain in dogs. Sensations of touch, proprioception, and high-threshold mechanosensitive nociception, as well as locomotor function, remained intact in both species. In separate experiments directed at postoperative pain control, subcutaneous administration of RTX transiently disrupted nociceptive nerve endings, yielding reversible analgesia. In human dorsal root ganglion cultures, RTX induced a prolonged increase in intracellular calcium in vanilloid-sensitive neurons, while leaving other, adjacent neurons unaffected. The results suggest that nociceptive neuronal or nerve terminal deletion will be effective and broadly applicable as strategies for pain management. PMID- 15124027 TI - Dysbindin-1 is reduced in intrinsic, glutamatergic terminals of the hippocampal formation in schizophrenia. AB - Eleven studies now report significant associations between schizophrenia and certain haplotypes of single-nucleotide polymorphisms in the gene encoding dysbindin-1 at 6p22.3. Dysbindin-1 is best known as dystrobrevin-binding protein 1 (DTNBP1) and may thus be associated with the dystrophin glycoprotein complex found at certain postsynaptic sites in the brain. Contrary to expectations, however, we found that when compared to matched, nonpsychiatric controls, 73-93% of cases in two schizophrenia populations displayed presynaptic dysbindin-1 reductions averaging 18-42% (P = 0.027-0.0001) at hippocampal formation sites lacking neuronal dystrobrevin (i.e., beta-dystrobrevin). The reductions, which were not observed in the anterior cingulate of the same schizophrenia cases, occurred specifically in terminal fields of intrinsic, glutamatergic afferents of the subiculum, the hippocampus proper, and especially the inner molecular layer of the dentate gyrus (DGiml). An inversely correlated increase in vesicular glutamate transporter-1 (VGluT-1) occurred in DGiml of the same schizophrenia cases. Those changes occurred without evidence of axon terminal loss or neuroleptic effects on dysbindin-1 or VGluT-1. Our findings indicate that presynaptic dysbindin-1 reductions independent of the dystrophin glycoprotein complex are frequent in schizophrenia and are related to glutamatergic alterations in intrinsic hippocampal formation connections. Such changes may contribute to the cognitive deficits common in schizophrenia. PMID- 15124028 TI - Neuronally expressed stem cell factor induces neural stem cell migration to areas of brain injury. AB - Neural stem/progenitor cell (NSPC) migration toward sites of damaged central nervous system (CNS) tissue may represent an adaptive response for the purpose of limiting and/or repairing damage. Little is known of the mechanisms responsible for this migratory response. We constructed a cDNA library of injured mouse forebrain using subtractive suppression hybridization (SSH) to identify genes that were selectively upregulated in the injured hemisphere. We demonstrate that stem cell factor (SCF) mRNA and protein are highly induced in neurons within the zone of injured brain. Additionally, the SCF receptor c-kit is expressed on NSPCs in vitro and in vivo. Finally, we demonstrate that recombinant SCF induces potent NSPC migration in vitro and in vivo through the activation of c-kit on NSPCs. These data suggest that the SCF/c-kit pathway is involved in the migration of NSPCs to sites of brain injury and that SCF may prove useful for inducing progenitor cell recruitment to specific areas of the CNS for cell-based therapeutic strategies. PMID- 15124029 TI - Noise minimization in eukaryotic gene expression. AB - All organisms have elaborate mechanisms to control rates of protein production. However, protein production is also subject to stochastic fluctuations, or "noise." Several recent studies in Saccharomyces cerevisiae and Escherichia coli have investigated the relationship between transcription and translation rates and stochastic fluctuations in protein levels, or more generally, how such randomness is a function of intrinsic and extrinsic factors. However, the fundamental question of whether stochasticity in protein expression is generally biologically relevant has not been addressed, and it remains unknown whether random noise in the protein production rate of most genes significantly affects the fitness of any organism. We propose that organisms should be particularly sensitive to variation in the protein levels of two classes of genes: genes whose deletion is lethal to the organism and genes that encode subunits of multiprotein complexes. Using an experimentally verified model of stochastic gene expression in S. cerevisiae, we estimate the noise in protein production for nearly every yeast gene, and confirm our prediction that the production of essential and complex-forming proteins involves lower levels of noise than does the production of most other genes. Our results support the hypothesis that noise in gene expression is a biologically important variable, is generally detrimental to organismal fitness, and is subject to natural selection. PMID- 15124030 TI - Exclusion spirometry: a good idea. PMID- 15124031 TI - Exclusion spirometry: a bad idea. PMID- 15124032 TI - Respirologists--doing what we can. PMID- 15124033 TI - Tuberculosis control in Ecuador: unforeseen problems, unanticipated strengths. PMID- 15124034 TI - Exclusion spirometry: an initiative to increase lung function assessment in primary care. AB - Ongoing spirometry quality standards are difficult to bring into the daily routine of general practice. As a result, spirometry is rarely performed by primary care physicians. A new approach is proposed: exclusion spirometry. Acceptable and reproducible results are sought. However, the goal of the test is to try to reach values within normal limits, even if results do not reach quality standards. Normal results would be sufficient to exclude respiratory impairment, except in asthma. Abnormal results would require further testing in a diagnostic spirometry laboratory. The aim of the initiative is to enhance the compliance of general practitioners in using spirometers for screening. PMID- 15124035 TI - Physiological changes at altitude in nonasthmatic and asthmatic subjects. AB - Exercised-induced asthma is not due to exercise itself per se, but rather is due to cooling and/or drying of the airway because of the increased ventilation that accompanies exercise. Travel to high altitudes is accompanied by increased ventilation of cool, often dry, air, irrespective of the level of exertion, and by itself, this could represent an 'exercise' challenge for asthmatic subjects. Exercise-induced bronchoconstriction was measured at sea level and at various altitudes during a two-week trek through the Himalayas in a group of nonasthmatic and asthmatic subjects. The results of this study showed that in mild asthmatics, there was a significant reduction in peak expiratory flow at very high altitudes. Contrary to the authors' hypothesis, there was not a significant additional decrease in peak expiratory flow after exercise in the asthmatic subjects at high altitude. However, there was a significant fall in arterial oxygen saturation postexercise in the asthmatic subjects, a change that was not seen in the nonasthmatic subjects. These data suggest that asthmatic subjects develop bronchoconstriction when they go to very high altitudes, possibly via the same mechanism that causes exercise-induced asthma. PMID- 15124036 TI - What is unique about the new Canadian COPD guidelines? PMID- 15124037 TI - Surfactant--where are we in 2003? PMID- 15124038 TI - Steroids and/or cytotoxic agents should be used early in the management of patients with IPF--the pro argument. PMID- 15124039 TI - Steroids and/or cytotoxic agents should be used early in the management of patients with IPF - the con argument. PMID- 15124040 TI - Helical computed tomography has a role in the screening of lung cancer--the pro argument. PMID- 15124041 TI - Helical computed tomography has a role in the screening of lung cancer--the con argument. PMID- 15124042 TI - Combination of ICSs and LABAs should be used in the management of patients with COPD--the pro argument. PMID- 15124043 TI - Combination of ICSs and LABAs should be used in the management of patients with COPD - the con argument. PMID- 15124044 TI - Recent advances in therapy for gastric cancer. AB - Gastric cancer is the fourth leading cause of death from cancer in Taiwan. Complete surgical resection is the only potentially curative treatment for gastric cancer. Randomized trials have failed to show the superiority of D2 over D1 dissection, and comparisons showing higher survival rates following more extensive surgery in Japan may have been influenced at least in part by the fact that D1 dissection underestimates disease stage in Western populations. No studies have shown convincing evidence of a survival benefit from adjuvant chemotherapy. A regimen of postoperative 5-fluorouracil (5-FU)-based chemoradiotherapy can improve disease-free and overall survival compared with surgery alone. Inadequate lymph node dissection for local control is a major concern. For advanced disease, HDFL (weekly 24-hour infusion of high-dose 5-FU and leucovorin)-based 'doublet' chemotherapy forms a cornerstone of combination chemotherapy in gastric cancer, but the effect on prolonged median survival is minimal. A 'sequential' non-cross-resistant strategy may be useful to prolong overall survival in patients with advanced gastric cancer. There are indications that preoperative neoadjuvant chemotherapy or chemoradiotherapy may increase the resectability of tumors and reduce the risk of postoperative recurrence. In the future, substantial improvements of treatment outcome will likely depend on the integration of novel, molecularly targeted agents into multimodality treatment strategies for all stages of gastric cancer. Further clinical research is mandatory to develop optimal therapies for gastric cancer. PMID- 15124045 TI - Relationship between Helicobacter pylori infection and erosive gastroesophageal reflux disease. AB - BACKGROUND AND PURPOSE: The role of Helicobacter pylori in the pathogenesis of gastroesophageal reflux disease remains controversial. The aims of the study were to assess the factors that influence the severity of erosive esophagitis in Taiwanese and to determine the prevalence of H. pylori infection in symptomatic patents. METHODS: Patients with erosive esophagitis confirmed by upper endoscopy from January 2002 to December 2002 at National Taiwan University Hospital were enrolled. The severity of erosive esophagitis was graded endoscopically according to the Los Angeles classification. H. pylori infection was determined by pathology and culture of biopsies at the gastric body and/or antrum. A sex matched control was selected to determine the prevalence of H. pylori infection. RESULTS: A total of 104 patients were recruited for the study, 3 of whom were excluded due to previous eradication of H. pylori infection. The prevalence of H. pylori infection in the 101 eligible patients with erosive esophagitis was 34.7%, which was significantly lower than the 64.9% prevalence in the control group (p < 0.001). Age and hiatal hernia were significantly associated with the severe erosive esophagitis (odds ratio, 1.05 and 4.44, respectively). H. pylori infection status, gender, smoking, alcohol drinking, and coexistent systemic diseases such as diabetes and hypertension were not correlated with the severity of erosive esophagitis. CONCLUSIONS: The overall prevalence of H. pylori infection was significantly lower in symptomatic patients with erosive esophagitis than in controls. Age and hiatal hernia were significantly associated with the severity of erosive esophagitis. H. pylori infection was inversely correlated with the occurrence of erosive esophagitis, but was not correlated with its severity. PMID- 15124046 TI - Simultaneous determination of indinavir, ritonavir and saquinavir in plasma by high-performance liquid chromatography. AB - BACKGROUND AND PURPOSE: Antiretroviral regimens consisting of a combination of protease inhibitors have been shown to be highly potent for the treatment of HIV infection. Therapeutic drug monitoring may ensure optimal drug levels to improve efficacy and minimize side effects. The purpose of this study was to establish a method for simultaneous determination of plasma concentrations of indinavir (IDV), ritonavir (RTV) and saquinavir (SQV) by high-performance liquid chromatography (HPLC) in a single run. METHODS: A C(18) column and an ultraviolet detector set at 215 nm were used in the HPLC. The mobile phase consisted of phosphate buffer (50 mM, pH 5.6) and acetonitrile (55:45, v/v), and the flow rate was 1.5 mL/min. Plasma sample (0.5 mL) containing IDV, RTV, and SQV was alkalinized with ammonia water (10%) and propylparaben added as an internal standard (IS), which was then extracted with methyl tert.-butyl ether. The organic layer was taken and evaporated to dryness. The residue was dissolved in the mobile phase, and the solution was washed with n-hexane followed by HPLC analysis. RESULTS: The calibration curves were linear for each of the 3 drugs in the studied concentration range (0.1 to 5 microg/mL). The extraction recoveries were greater than 90%. One HPLC run takes less than 15 minutes. Analysis of plasma samples of patients treated with combinations of these drugs demonstrated that the calibration curves covered the clinical concentration range. CONCLUSIONS: This study established a simple and accurate method for simultaneous determination of IDV, RTV, and SQV in plasma by isocratic HPLC. The method is practical for therapeutic drug monitoring. PMID- 15124047 TI - Health-related quality of life among urban, rural, and island community elderly in Taiwan. AB - BACKGROUND AND PURPOSE: There are published norms of the 36-item short form of the Medical Outcomes Study Questionnaire (SF-36) for many countries, but few such studies have been conducted in Taiwan. The purpose of this study was to provide the norms for the SF-36 health status measure in urban, rural, and remote island community elderly populations, and to explore the relation between demographic or characteristic factors and the SF-36. METHODS: A structured questionnaire was used for door-to-door data collection in this survey of an urban, a rural, and a remote population (Shihpai, Yuli, and Kinmen, respectively, in Taiwan). Interviewers also collected information on subjects' demographics, medical history, use of health services, and health-related quality of life using the SF 36. A total of 6503 subjects who were at least 65 years of age were invited, and 4424 (73.1%) participated in the survey. RESULTS: The urban norms were significantly higher than the rural norms in seven scales of the SF-36 and significantly higher than the remote island norms in 4 scales, including physical functioning (84.6 vs 77.1), role limitations due to physical problems (77.8 vs 70.2), general health perceptions (70.5 vs 65.5), and role limitations due to emotional problems (90.6 vs 85.2). Remote island norms were significantly higher than urban and rural norms in vitality and mental health scales. In the multivariate analyses, women had significantly poorer scores in all scales of the SF-36 than men (p < 0.05), except for social functioning. Higher education was significantly associated with increased scores in all scales, except for vitality/energy and mental health scales. Scores of all scales were significantly reduced according to the number of chronic diseases a subject had. The need for domestic services and utilization of inpatient and outpatient health services were associated with lower scales (p < 0.05). CONCLUSIONS: There were significant differences between urban, rural, and remote island elderly populations in most scales of the SF-36. The urban elderly population had the greatest health-related quality of life on most scales, particularly on the physical health scales. The remote island elderly population had the highest scores on the vitality and mental health scales, whereas the rural elderly population had the poorest health related quality of life, particularly rural women. PMID- 15124048 TI - Health-related quality of life in Taiwanese patients with bronchial asthma. AB - BACKGROUND AND PURPOSE: Health-related quality of life (HRQOL) is primarily an assessment of how domains of life are affected by health. This study investigated the factors influencing HRQOL of Taiwanese patients with bronchial asthma. METHODS: A cross-sectional study was conducted to assess the factors influencing HRQOL. The sample consisted of 242 outpatients with bronchial asthma. The Chinese language version of the St. George's Respiratory Questionnaire was used to measure HRQOL. Data were analyzed using simple regression and multiple regression. RESULTS: Age, marital status, level of education, asthma severity, peak expiratory flow, and previous hospital admissions were found to be predictors of HRQOL. Gender, duration of disease, and history of emergency visits were not correlated with HRQOL. The variables entered in the HRQOL model accounted for 17.4% of the total variance (adjusted R(2)). The regression coefficients indicated that the mean scores increased 7.68 in patients with moderate to severe disease severity, and increased 7.34 in patients with a history of hospital admissions. The mean scores decreased 7.60 in married patients. CONCLUSION: This study found that marital status, asthma severity, and history of hospital admissions were major predictors of HRQOL in Taiwanese patients with bronchial asthma. PMID- 15124049 TI - Visual outcome in primary macula-off rhegmatogenous retinal detachment treated with scleral buckling. AB - BACKGROUND AND PURPOSE: To evaluate the visual outcome of primary macula-off rhegmatogenous retinal detachment after successful scleral buckling. METHODS: A retrospective, non-controlled case series study was conducted in 93 patients (93 eyes) who underwent primary successful scleral buckling procedure for retinal detachment. Factors including duration of macular detachment, patient age, preoperative best-corrected visual acuity (VA), surgical management of subretinal fluid, and refractive error were analyzed statistically to determine their association with final visual outcome. RESULTS: Postoperative VA of 20/50 or better was found in 53.6% of eyes with duration of macular detachment within 7 days, and 29.7% of eyes with macular detachment for more than 7 days (Fisher's exact test, p = 0.019). VA better than 20/50 was found in 61% of eyes with preoperative VA better than 20/400 and in 33.9% with preoperative VA worse than 20/400 (Fisher's exact test, p = 0.008). Patients aged 30 years or less achieved better mean postoperative VA than those aged 31 to 50 and those aged 50 years and older (ANOVA, p = 0.003). Patients with low-grade myopia (< -6D) regained significantly better mean postoperative VA as compared with high myopia (> -6D) and emmetropic eyes (0 to +3D) (ANOVA, p < 0.001). Subretinal fluid drainage procedure did not affect postoperative visual result. Multivariate logistic regression analysis revealed that the duration of macular detachment was the only variable affecting the visual result. CONCLUSION: Scleral buckle surgery performed within the first week, preoperative vision more than 20/400, age younger than 30 years old, and low-grade myopia were associated with significantly better visual recovery from macular-off rhegmatogenous retinal detachment. PMID- 15124050 TI - Cytogenetic surveillance of mentally-retarded school children in southern Taiwan. AB - BACKGROUND AND PURPOSE: Mental retardation (MR), defined as having an IQ of less than 70, is present in approximately 2 to 3% of the population. Data on chromosomal abnormalities, an important cause of MR, are limited in the Taiwanese literature. This study evaluated the frequency and pattern of chromosomal abnormalities in school children with MR in southern Taiwan. METHODS: Peripheral blood samples of 419 children were collected from November 1999 to January 2003. Those with Prader-Willi syndrome (PWS), Angelman syndrome (AS) or fragile-X syndrome were excluded from the study. Metaphase chromosome preparations were obtained from peripheral blood cultures, and trypsin-giemsa (GTG) banded chromosomes were examined at the level of 500 to 600 bands. Fluorescence in situ hybridization was done for cases whose karyotypes could not be determined by conventional cytogenetic analysis. RESULTS: Of the 419 enrollees with MR, 10 had mild MR, while most had moderate to profound MR. Chromosomal abnormalities were found in 22.43% of the cases, with trisomy 21 being the major chromosomal abnormality, occurring in 77 cases (18.38%, 77/419 cases). Sex chromosome aneuploidies were found in 3 cases (0.72%, 3/419 cases). Structural abnormalities of autosomes were found in 13 cases (3.10%, 13/419 cases), including deletion, markers, unbalanced translocations, and inversions. One subject was found to have monosomy 20 mosaicism (0.24%, 1/419). CONCLUSIONS: Chromosomal abnormalities occurred in a high proportion of mentally retarded school children from southern Taiwan, with trisomy 21 being the most prevalent. These findings indicate the need for increased attention to prenatal, perinatal and postnatal screening in this population. PMID- 15124052 TI - Intraspinal extradural ganglion cyst of the cervical spine. AB - Intraspinal ganglion cyst is an uncommon lesion, which occurs most frequently in the lower lumbar region; occurrence in the cervical region is extremely rare. We report a case of ganglion cyst in the cervical region and describe its clinical pathological and radiological findings. A 59-year-old man presented with sudden lower limbs weakness and numbness below the nipple level. Magnetic resonance imaging revealed an intraspinal extradural lobulated cystic lesion at the level of C6-7 in contact with the left facet joint and posterior erosion of the spinal process at C7. The spinal cord was severely compressed by this lesion which was hypointense on T1-weighted imaging and hyperintense on T2-weighted imaging and short T1 inversion recovery. The cyst wall was strongly enhanced after contrast injection. Intraoperatively, the mass was found to arise from the capsule of the C6-7 facet joint. The excised cyst contained jelly-like fluid. The patient's neurologic symptoms had fully recovered 20 days after the operation. The histopathologic analysis was consistent with ganglion cyst. Ganglion cyst of the cervical region is extremely rare, but must be considered in the differential diagnosis of intraspinal extradural compressive syndromes. PMID- 15124051 TI - E207K mutation of low-density lipoprotein receptor in familial hypercholesterolemia. AB - We report a case of heterozygous familial hypercholesterolemia (HeFH) in a 36 year-old man with premature coronary artery disease (CAD). Hypercholesterolemia was found in family members including his mother, wife and all 3 of his children (2 boys aged 6 and 3 years, 1 girl aged 4 years). Genetic analysis revealed a G- >A substitution at nucleotide 682, resulting in Glu(207) to Lys (E207K) mutation of the ligand-binding domain of the low-density lipoprotein receptor (LDLR) of all the family members with hypercholesterolemia except for the proband's wife. Genetic study showed that this mutation was inherited from the proband's mother then transmitted to all 3 children. Detection of this mutation identifies the cause of hypercholesterolemia and allows appropriate early treatment to prevent premature CAD. PMID- 15124053 TI - Femoral head preservation in non-united femoral neck fracture and head osteonecrosis in Cushing's disease. AB - There have been few reports associating avascular necrosis of bone with Cushing's disease. Patients with Cushing's disease and avascular necrosis of the femoral head usually receive total hip arthroplasty. However, hip prosthetic replacement in younger patients has been criticized due to a high incidence of component loosening. We report a case of successful femoral head preservation in non-united femoral neck fracture and head osteonecrosis in a 14-year-old girl with Cushing's disease (adrenocorticotropic hormone-secreting pituitary adenoma) who developed avascular necrosis of the right femoral head and pathologic fracture of the right femoral neck 2 years after the onset of hypercortisolism. Subtrochanteric valgus osteotomy was performed to preserve the femoral head after successful transsphenoidal surgery to remove pituitary microadenoma. At follow-up 10 years after the osteotomy, the femoral head had revascularized and the femoral neck fracture were united with much improvement of hip function. Dual energy X-ray absorptiometry scan of the right hip showed +0.86 SD from the normal bone densitometry. Aggressive femoral head preservation may be an effective alternative to treat this rare situation in a teenager. PMID- 15124054 TI - Partial chromosomal 11q trisomy as an unbalanced result of t(11;13)(q23.1;q34)mat in two sisters. AB - Partial trisomy of chromosome 11q is a rare but rather distinct clinical syndrome. Cases of distal trisomy of chromosome 11(q23.1-qter) in 2 sisters are described. The features included microbrachycephaly, long philtrum, retracted lower lip, short neck, heart defects, marked leukocytosis, and psychomotor retardation. These patients died at the age of 3 months and 6 months, respectively. Chromosome analyses showed a distal trisomy 11q resulting from maternal t(11;13)(q23.1;q34). Fluorescence in situ hybridization using the painting probes for chromosomes 11 and 13, along with a unique sequence for mixed lineage leukemia (MLL) gene confirmed this condition. The phenotypes of both sisters are most likely to be related to partial trisomy of 11q and a triplicated gene dosage of MLL. PMID- 15124055 TI - Buccal mucosal graft for urethral reconstruction. AB - Urethral reconstruction with graft substances, such as skin and bladder mucosa, has been previously used when primary anastomosis cannot be achieved. However, stricture and meatal prolapse are associated with these grafts. We report the use of buccal mucosa for the reconstruction of urethral defects in 3 patients. One patient with failed operation for hypospadias received tube buccal mucosal graft for urethral replacement. Two patients with urethral necrosis and stricture received onlay buccal mucosal graft. All patients were disease-free during follow up (range, 12 to 49 months; mean, 36 months). One patient had a pinhole fistula that was successfully managed with simple repair. This technique appears to be useful for urethral reconstruction when a local graft is not available, even in patients with complicated conditions. PMID- 15124068 TI - [International severity scale of diabetic retinopathy and diabetic macular edema]. PMID- 15124069 TI - [Liposomes as ocular drug delivery systems]. PMID- 15124070 TI - [Post-operative endophthalmitis. Developing guidelines for good clinical practice]. PMID- 15124071 TI - [Corneal epithelial complications after vitrectomy: a retrospective study]. AB - PURPOSE: To establish the incidence, evolution and final visual outcome of corneal epithelial complications after vitrectomy in a consecutive series performed throughout a period of one year. METHODS: 195 consecutive medical records of patients vitrectomized in 2001 have been retrospectively reviewed. Patients were clinically evaluated the day after vitrectomy, during the first week, and then weekly for 3 months (minimum follow-up). Functional results have been established at the end of this period. RESULTS: 12 out of 195 eyes developed corneal epithelial complications after surgery (6.1%). Eight cases were diagnosed of persistent epithelial defect (4.1%), and four cases (2%) of necrotizing herpetic keratitis. Six out of 12 were diabetic. Corneal epithelial complications affected 10.5% of diabetic and 4.3% of non-diabetic patients (p=0.10). The majority of persistent epithelial defects healed without sequelae in less than two months. Herpetic keratitis required between 3 to 4 months to heal and caused corneal scarring in all cases. CONCLUSIONS: Persistent corneal epithelial defects after vitrectomy are a relatively frequent complication, mainly in diabetic patients. Recurrence of herpetic keratitis must also be kept in mind. This finding has not been previously reported and needs further studies. PMID- 15124072 TI - [Efficiency of brimonidine 0.2% and dorzolamide 2% as adjunctive therapy to beta blockers]. AB - PURPOSE: To evaluate the clinical efficiency and tolerability of brimonidine and dorzolamide twice daily as an adjunctive therapy for glaucoma patients with an inadequate response to beta-blockers therapy. METHODS: This multicenter prospective analysis included 92 patients (180 eyes) with primary open-angle glaucoma or ocular hypertension on therapy beta-blockers and with intraocular pressure (IOP) greater than or equal to 18mmHg. The patients were randomly treated either with brimonidine 0.2% or dorzolamide 2% added for three months. Efficiency was determined by the reduction in 15% IOP from baseline at the first and the third month. RESULTS: Mean pre-treatment IOP was 22.37 DE 2.8 mmHg in the brimonidine group and 22.38 DE 2.6 mmHg in the dorzolamide group; mean post treatment IOP decrease was 4.39 mmHg in the brimonidine group and 3.29 mmHg in the dorzolamide group. Clinical control at the first month was achieved in 78.3% and 71% of cases respectively (p=0.05). No statistical differences existed between groups for systemic adverse events. Four patients on brimonidine discontinued treatment due to local side effects. In the dorzolamide group, two patients left the treatment referring itching and three others left due to ocular allergy. CONCLUSIONS: This study found similar efficiency and safety when treating with brimonidine or dorzolamide as an adjunctive therapy for patients with hypertension or primary open-angle glaucoma. PMID- 15124073 TI - [A study of lacunar folds in relation to the etiology and severity of dry eye]. AB - PURPOSE: To determine the diagnostic value of studying lacunar folds in relation to the different etiologies responsible for dry-eye syndrome. METHODS: We present a prospective study of 206 eyes from 106 patients (38 males and 68 females). In all patients the abduction degree was measured after instilling a fluorescein drop, which makes lacunar folds disappear. We divided the patients into ten etiological groups applying Madrid classification criteria. Patients were also divided into four groups according to clinical severity. RESULTS: We found the early disappearance of lacunar folds was more prevalent among patients with immunological dry eye. Early disappearance was less frequent in patients with tantalic, pharmacological, inflammatory and traumatic dry eye. The folds disappeared more rapidly in patients with greater clinical severity. CONCLUSIONS: The early disappearance of lacunar folds is a sign of conjunctival retraction manifest in patients with dry eye syndrome. The degree of conjunctival retraction degree varies according to the dry eye etiology and severity of the symptoms. PMID- 15124074 TI - [Serous retinal detachment in systemic erithematosus lupus during corticosteroid therapy]. AB - CASE REPORT: A young female patient with systemic lupus erithematosus, inactive for a long period, is described. She develops bilateral serous detachments of the retinal pigment epithelium and neurosensory retina after corticosteroid therapy for a residual nephritis. DISCUSSION: This unusual chorioretinopathy has been reported in association with systemic lupus erithematosus and during corticosteroid therapy in many disorders. This patient, with low activity erithematosus lupus, develops serous retinal detachments after receiving corticosteroid therapy that damage the coriocapillary-pigment epithelium barrier, disordered in lupus patients. PMID- 15124075 TI - [Oral itraconazole for treatment of a candida parapsilosis endophthalmitis case]. AB - CLINICAL CASE: A case of exogenous Candida parapsilosis endophthalmitis was treated first with oral fluconazol and later with intravenous Anfotericin without success. A subsequent therapy using topical fluconazole and oral itraconazole appeared to be effective in controlled the infection. DISCUSSION: Current elective treatment for fungal endophthalmitis is Anfotericine B i.v. An alternative therapy are the azoles. In the reported case itraconazole was used, although its intraocular penetration and effectiveness have not yet been satisfactorily demonstrated. The clinical response in this case leads to the possibility that intraocular oral itraconazole penetration can be sufficient to control infection in cases where the causal fungi is sensitive. PMID- 15124076 TI - [Bilateral traumatic retinopathy associated to whiplash injury]. AB - CASE REPORT: A 20-year old female complained of a paracentral scotoma after having suffered a whiplash injury. Eye fundus examination and fluorescein angiography suggested the diagnosis of traumatic retinopathy. DISCUSSION: There is not a clear explanation to justify the development of traumatic retinopathy after an indirect ocular trauma. Some authors favour a mixed mechanism, mechanic and vascular, to explain these alterations. We consider that a vascular aetiology is the most possible explanation for the development of traumatic retinopathy. PMID- 15124077 TI - [Comments on Cogan's syndrome (congenital oculomotor apraxia) based on a case]. AB - CASE: A 4-month-old girl was seen by a paediatrician for psychomotor delay and defective visual fixation. In order to shift gaze direction, she had to jerk her head in order to drag her eyes in the direction she wanted to look. She also showed non-specific pyramidal impairment. Herpes human virus type 6 (HHV-6) was amplified by PCR. No other findings could explain her troubles with fixation. She was diagnosed with congenital ocular motor apraxia (C.O.M.A.). DISCUSSION: When C.O.M.A. is suspected, other causes of defective fixation and abnormal cephalic movements must be ruled out through the use of physical examination, electrophysiology and neuroimaging: troubles with ocular motility, poor vision or neurological processes. There have been cases associated with neurological misdevelopment, systemic diseases, metabolic deficits and chromosomic abnormalities. PMID- 15124078 TI - [Ophthalmology for the rural physician (1919)]. PMID- 15124080 TI - Prevention of experimentally induced irritant contact dermatitis by extracts of Isatis tinctoria compared to pure tryptanthrin and its impact on UVB-induced erythema. AB - Lipophilic extracts of Isatis tinctoria L. exhibit significant activity against several clinically relevant targets of inflammation. The alkaloid tryptanthrin was identified as one of the active principles in woad and characterised as a potent dual inhibitor of COX-2 and 5-LOX. Here, the anti-inflammatory efficacy of topical application of three different Isatis extracts and tryptanthrin was investigated in human volunteers. Two different models were used, namely the sodium lauryl sulphate (SLS)-induced irritant contact dermatitis (ICD) and UVB induced erythema. Twenty healthy volunteers without any skin disease participated in the study. Cumulative irritant contact dermatitis was induced on test fields on the volunteers' backs by twice daily application of 0.5 % sodium lauryl sulphate over a period of four days. Half of the test fields were treated with the test substances during the eliciting phase, while the remaining test fields were treated over a period of 4 days after induction of dermatitis. In the second model, a UVB erythema on the volunteers' lower backs was induced using the double minimal erythema dose (MED). Twenty-four hours after irradiation the test fields were treated with the test substances over a period of 3 days. All reactions were assessed visually and by non-invasive bioengineering methods (evaporimetry and chromametry). Treatment with extracts during the ICD eliciting phase led to a significantly smaller increase of visual scores and transepidermal water loss compared to the untreated test field. For tryptanthrin this benefit was also observed, but the improvement was not statistically significant. When treatment was performed after completing the eliciting phase, accelerated resolution of the irritant reaction could not be observed. In the UVB erythema model anti inflammatory effects of the test substances were not observed. PMID- 15124081 TI - Aristolactam BII of Saururus chinensis attenuates glutamate-induced neurotoxicity in rat cortical cultures probably by inhibiting nitric oxide production. AB - Saurolactam and aristolactam BII, aristolactam-type alkaloids isolated from the aerial part of Saururus chinensis (Lour.) Ball (Saururaceae), showed significant neuroprotective activity against glutamate-induced toxicity in primary cultured rat cortical cells. The action mechanism of aristolactam BII, the more potent neuroprotective compound, was investigated using primary cultures of rat cortical cells as an in vitro system. Aristolactam BII attenuated glutamate-induced neurotoxicity significantly when it was added immediately or up to 9 h after the excitotoxic glutamate challenge. The alkaloid could not protect cultured neuronal cells from neurotoxicity induced by kainic acid or N-methyl- D-aspartate in a pre treatment paradigm. However, aristolactam BII successfully reduced the overproduction of nitric oxide and the level of cellular peroxide in cultured neurons when it was treated as a post-treatment paradigm. These results may suggest that aristolactam BII exerts its significant neuroprotective effects on glutamate-injured primary cultures of rat cortical cells by directly inhibiting the production of nitric oxide. PMID- 15124082 TI - Silibinin down-regulates prostate epithelium-derived Ets transcription factor in LNCaP prostate cancer cells. AB - The androgen-sensitive human prostate cancer cell line LNCaP expresses the estrogen receptor beta and androgen receptor and can be stimulated by androgens to secrete prostate-specific antigen (PSA). In this study we demonstrate the cancer protective potential of silibinin, a flavolignan derived from the fruits of Silybum marianum, which down-regulates the co-activator of the androgen receptor, the prostate epithelium-derived Ets transcription factor (PDEF) and consequently the secretion of PSA. LNCaP cells were treated with various concentrations of silibinin in the presence or in the absence of 5alpha dihydrotestosterone (DHT). We used real-time RT-PCR to quantify mRNA expression of PDEF and PSA with gene-specific dual-labelled fluorescence probes. PSA secretion from LNCaP cells in conditioned media was measured with the Elecsys System 2010. Silibinin down-regulated PSA mRNA expression and PSA secretion in conditioned medium under basal and DHT (10(-8) M) stimulated conditions, which was paralleled by PDEF down-regulation. DHT alone stimulated PDEF and PSA gene expression and PSA secretion. The down-regulation of basal as well as DHT stimulated PDEF and PSA by silibinin demonstrates the antiproliferative potential of this agent. These effects underline the possible therapeutic use of silibinin in the management of prostate cancer. PMID- 15124083 TI - A comparison of apoptosis and necrosis induced by ent-kaurene-type diterpenoids in HL-60 cells. AB - We previously reported that ent-11alpha-hydroxy-16-kauren-15-one (KD) induces apoptosis through a caspase-dependent pathway and the induction of apoptosis is dependent on its enone group in human leukemia cells. Here we investigated the abilities of some KD-related compounds with enone group (Fig. 1) to induce apoptosis and to activate some caspases. The IC50 values of ent-kaurene-related compounds possessing the enone group, ent-1beta-hydroxy-9(11),16-kauradien-15-one (1), ent-9(11),16-kauradiene-12,15-dione (2) and the rearranged ent-kaurane-type diterpene (3), against HL-60 cells after 12 h of treatment were 40 microM, 1.8 microM and 5.5 microM, respectively. Although treatment with 3 induced apoptosis, DNA ladder formation was not observed after treatment with 1 or 2. Induction of necrosis, as assayed by trypan blue staining, was observed after treatment with 1 or 2. Treatment with compound 1, 2 or 3 induced proteolysis of poly(ADP-ribose) polymerase (PARP), a substrate of caspase-3, and processing of caspase-3. Activation of caspase-8 and processing of Bid, a typical substrate of caspase-8, were also observed on treatment with these compounds. Pretreatment with a broad spectrum inhibitor of caspases attenuated apoptosis induced by 3 but not necrosis induced by 1 and 2. In summary, KD-related compounds are a unique family of diterpenes that cause either caspase-dependent apoptotic or necrotic cell death. PMID- 15124084 TI - Alkaloids from Cassytha filiformis and related aporphines: antitrypanosomal activity, cytotoxicity, and interaction with DNA and topoisomerases. AB - Cassytha filiformis (Lauraceae), a widely distributed parasitic plant, contains several aporphine alkaloids and is often used in African folk medicine to treat cancer, African trypanosomiasis and other diseases. In a previous investigation, we showed that the alkaloid plant extract and the isolated aporphines possessed in vitro cytotoxic properties. In this paper, we evaluated the in vitro activity of the alkaloid extract (IC50 = 2.2 microg/mL) and its three major aporphine alkaloids (actinodaphnine, cassythine, and dicentrine) on Trypanosoma brucei brucei as well as four related commercially available aporphines (bulbocapnine, glaucine, isocorydine, boldine). Only the three alkaloids from Cassytha filiformis were active on the trypanosomes in vitro (IC50 = 3-15 microM). Additionally, we compared the cytotoxicity of these seven compounds on HeLa cells. Glaucine was the most cytotoxic compound on HeLa cells (IC50 = 8.2 microM) in the series. In order to elucidate their mechanism of action, the binding mode of these molecules to DNA was studied by UV absorption, circular and linear dichroism spectroscopy. The results of the optical measurements indicated that all seven aporphines effectively bind to DNA and behave as typical intercalating agents. Biochemical experiments showed that actinodaphnine, cassythine and dicentrine also interfere with the catalytic activity of topoisomerases in contrast to the four other aporphines. These interactions with DNA may explain, at least in part, the effects observed on cancer cells and on trypanosomes. PMID- 15124085 TI - Anti-inflammatory activity of two cucurbitacins isolated from Cayaponia tayuya roots. AB - Fractionation of an anti-inflammatory extract from Cayaponia tayuya roots yielded two active compounds, identified as 23,24-dihydrocucurbitacin B (1) and cucurbitacin R (2). Both were evaluated for their anti-inflammatory activity on several experimental models of pain and inflammation. In addition, their cytotoxicity and effects on leukotriene B4 (LTB4) formation were evaluated in rat polymorphonuclear leukocytes. Both compounds showed activity in the following models: carrageenan-induced mouse paw oedema (1, 4 mg/kg p.o., 46% inhibition at 3 h), phospholipase A2-induced mouse paw oedema (2, 3 mg/kg i.p., 61% inhibition at 60 min), serotonin-induced mouse paw oedema (1 and 2, 0.5 mg/kg s.c., 73% and 79% inhibition, respectively), 12- O-tetradecanoylphorbol 13-acetate (TPA) induced acute ear oedema (2, 36% inhibition at 4 mg/kg p.o., and 87% inhibition at 0.1 mg/ear topically). The compounds were also active against the inflammation induced by repeated application of TPA on mouse ears, affecting both the oedema itself (1 and 2 at 0.1 mg/ear, 44% and 56% inhibition, respectively) as well as cell infiltration (68% and 69%, respectively). The activity of both compounds against oedema induced by serotonin was not modified by the glucocorticoid receptor antagonist mifepristone; however, the protein synthesis inhibitor cycloheximide abolished the anti-inflammatory response in both cases. Neither compound modified the production of LTB4 in rat polymorphonuclear leukocytes, nor did they exhibit analgesic properties at the dose assayed. PMID- 15124086 TI - A new anti-inflammatory glucoside from Ficus racemosa L. AB - Bioassay-guided fractionation of the ethanol extract of Ficus racemosa resulted in the identification of a new compound (rel)-4,6-dihydroxy-5-[3-methyl-(E) propenoic acid-3-yl]-7-beta-glucopyranosyl-[2alpha,3beta-dihydrobenzofuran]-(3,2: b)-[4alpha,5beta-dihydroxy-6alpha-hydroxymethyltetrahydropyran] (racemosic acid). Racemosic acid showed potent inhibitory activity against COX-1 and 5-LOX in vitro with IC50 values of 90 and 18 microM, respectively. Racemosic acid also demonstrated a strong antioxidant activity to scavenge ABTS free radical cations with an IC50 value of 19 microM. In addition, cytotoxic effects of the extracts of F. racemosa were investigated in vitro using the ATP-based luminescence assay and results showed no cytotoxicity on the cell lines skin fibroblasts (1BR3), human Caucasian hepatocyte carcinoma (Hep G2) and human Caucasian promyelocytic leukaemia (HL-60). Bergenin was also isolated from the same active fraction. PMID- 15124087 TI - Scutellarin attenuates oxidative glutamate toxicity in PC12 cells. AB - The present study investigated the protective effects of the antioxidant scutellarin against oxidative toxicity induced by glutamate in PC12 cells. Vitamin E, a classical antioxidant was employed as a comparative agent. Incubation of PC12 cells with 10 mM glutamate resulted in significant cytotoxity as evaluated by the MTT and lactate dehydrogenase (LDH) assays, decreases of GSSG reductase activity, disturbance of the cell redox state as indicated by the GSH/GSSG ratio, and accumulation of intracellular reactive oxygen species (ROS) and lipid peroxidation products. Scutellarin at 0.1, 1 and 10 microM significantly protected against the cytoxicity and production of ROS and lipid peroxidation induced by glutamate. Scutellarin did not prevent the reduction of cellular GSH levels, but it up-regulated GSSG reductase activity, thus preventing an increase in cellular GSSG levels, and concomitantly improved the cell redox status. Our data also show that the protective effects of scutellarin against glutamate-induced oxidative toxicity are more potent than that of vitamin E. These results demonstrate that scutellarin can protect PC12 cells from oxidative glutamate toxicity by scavenging ROS, inhibiting lipid peroxidation and improving the cell redox status, and may reduce the cellular damage in pathological conditions associated with excessive glutamate release. PMID- 15124088 TI - Stability control of senna leaves and senna extracts. AB - Powdered senna leaves and a commercial methanolic senna leaf extract were investigated for apparent degradation pathways of known constituents. Different defined storage conditions were chosen according to the guidelines of the international conference on harmonization. Analytical fingerprinting was carried out by HPLC with photodiode array detection. Differences in degradation pathways were observed between the powdered herbal drug material and the extract, depending on storage conditions and packaging materials. Within the crude plant material sennosides were shown to be degraded to sennidine monoglycosides, while rhein 8-O-glucoside was hydrolysed to rhein by enzymatic processes. Degradation of the anthranoid compounds was not due to the same pathways in the investigated commercial extracts. Only unspecific alterations of all compounds were observed. Forced decomposition of this herbal drug preparation under high temperature caused oxidative decomposition of the sennosides to rhein 8-O-glucoside. Furthermore flavonoid glycosides decomposition were observed with an apparent increase in the content of flavone aglyca. PMID- 15124089 TI - Antiplasmodial agents from the leaves of Glossocalyx brevipes. AB - A phytochemical study of the methylene chloride/methanol (1/1) extract of the leaves of Glossocalyx brevipes Benth. (Monimiaceae) afforded three new derivatives of homogentisic acid, methyl 2-(1'beta-geranyl-5'beta-hydroxy-2' oxocyclohex-3'-enyl)acetate (1), 2-(1'beta-geranyl-5'beta-hydroxy-2'-oxocyclohex 3'-enyl)acetic acid (2), methyl 2-(1'beta-geranyl-5'beta-hydroxy-4'beta-methoxy 2'-oxocyclohexyl)acetate (3), and two known alkaloids, aristololactam BII and liriodenine. Compounds 1 and 2 and liriodenine showed modest in vitro activity against Plasmodium falciparum. PMID- 15124090 TI - Cytotoxic alkaloids from the roots of Tylophora atrofolliculata. AB - Four new phenanthroindolizidine alkaloids, tylophoridicines C-F, together with three known ones, R-(+)-deoxytylophorinidine, tylophorinine and tylophorinidine, were isolated from the roots of Tylophora atrofolliculata. The structures were determined on the basis of spectral evidence. These alkaloids exhibited cytotoxic activity in vitro on HCT-8 cell (with IC50 values in the range 0.083 to 18.99 microM) and KB cell (in the range 3.56 to 18.22 microM) lines. PMID- 15124091 TI - Nor-lignans and steroidal saponins from Asparagus gobicus. AB - From the roots of Asparagus gobicus, four new nor-lignans, 3'-methoxynyasin, iso agatharesinol, gobicusins A, B and one new steroidal saponin, 3-O-[beta-D xylopyranosyl(1-4)-beta-D-glucopyranosyl(1-2)-beta-D-glucopyranosyl]-(25S)-5beta spirostan-3beta-ol (11) were isolated, together with twelve known compounds. The structures of the new compounds were established by spectroscopic methods including 2D-NMR techniques (1H-1H COSY, HMBC, HMQC) and chemical transformations. Nyasol (5) and 11 exhibited remarkable in vitro cytotoxic activity against cultured HO-8910 (human ovarian carcinoma) and Bel-7402 (human hepatoma) cells with IC50 vales of 30.6 and 29.4 microM, 5.2 and 5.2 microM, respectively. PMID- 15124092 TI - Diterpenoids from Salvia wagneriana. AB - From the surface exudate of the aerial parts of Salvia wagneriana, three new clerodane diterpenoids, the known hardwickiic acid and 1,10-didehydrosalviarin, were obtained. Two were bis-diterpenoids. Their structures were established by 1D and 2D-NMR spectroscopic techniques. PMID- 15124093 TI - New triterpenoid saponins from bulbs of Bolbostemma paniculatum. AB - Nine new triterpenoid saponins were isolated from the bulbs of Bolbostemma paniculatum (Maxim.) Franquet (Cucurbitaceae): 7beta,18,20,26-tetrahydroxy-(20S) dammar-24 E-en-3-O-alpha-L-(3-acetyl)arabinopyranosyl-(1-->2)-beta-D glucopyranoside, 7beta,18,20,26-tetrahydroxy-(20S)-dammar-24E-en-3-O-alpha-L-(4 acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,18,20,26 tetrahydroxy-(20S)-dammar-24E-en-3-O-alpha-L-arabinopyranosyl-(1-->2)-beta-D-(6 acetyl)glucopyranoside, 7beta,20,26-trihydroxy-(20S)-dammar-24E-en-3-O-alpha-L arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,20,26-trihydroxy-(20S) dammar-24E-en-3-O-alpha-L-(3-acetyl)arabinopyranosyl-(1-->2)-beta-D glucopyranoside, 7beta,20,26-trihydroxy-(20S)-dammar-24E-en-3-O-alpha-L-(4 acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside, 7beta,20,26-trihydroxy-8 formyl-(20S)-dammar-24E-en-3-O-alpha-L-(3-acetyl)arabinopyranosyl-(1-->2)-beta-D glucopyranoside, 7beta,20,26-trihydroxy-8-formyl-(20S)-dammar-24E-en-3- O-alpha-L (4-acetyl)arabinopyranosyl-(1-->2)-beta-D-glucopyranoside and 6'-O palmitoyltubeimoside I. In addition, four known triterpenoid saponins: tubeimoside I, tubeimoside II, tubeimoside III and tubeimoside IV were isolated. The structures of the above compounds were elucidated based on spectroscopic studies, and the configuration of C-20 of tubeimoside IV was revised as S rather than R as reported in previous literature. The compounds were tested for their antiviral activity PMID- 15124094 TI - Mechanisms involved in the vasodilator effect of the flavanol floranol in rat small mesenteric arteries. AB - The mechanism underlying the vasodilator effect of the flavonoid floranol was studied in rat small mesenteric arteries. Floranol produced a concentration dependent vasorelaxant effect in endothelium-containing and endothelium-denuded vessels pre-contracted with phenylephrine, which was more potent in endothelium intact vessels. In endothelium-intact mesenteric arteries, l-NAME but not indomethacin produced a shift to the right in the vasorelaxant effect of floranol. In endothelium-denuded vessels TEA and BaCl2 did not change the floranol-induced vasorelaxation. When endothelium-denuded vessels were pre contracted with 50 mM KCl, floranol induced a vasorelaxant effect comparable with phenylephrine pre-contracted vessels. We conclude that floranol is a new vasodilator compound in rat small mesenteric arteries. Part of this effect is dependent on endothelial nitric oxide (NO) and part is dependent on the inhibition of voltage-dependent calcium channels in the smooth muscle cells. PMID- 15124095 TI - Geniposide, an anti-angiogenic compound from the fruits of Gardenia jasminoides. AB - The EtOH extract of gardenia (Gardenia jasminoides Ellis) fruits was previously found to possess potent anti-angiogenic activity in the chick embryo chorioallantoic membrane (CAM) assay. Bioassay-guided fractionation and purification of the EtOH extract yielded an active anti-angiogenic compound, which was determined to be an iridoid glucoside, geniposide, by spectral analyses. Geniposide showed anti-angiogenic activity in a dose-dependent manner. It also exhibited an inhibitory effect in the range of 25-100 microM on the growth of the transformed NIH3T3 cell line. PMID- 15124096 TI - Production of aralin, a selective cytotoxic lectin against human transformed cells, in callus culture of Aralia elata. AB - Recently we found that aralin, a novel lectin, from Aralia elata selectively induces apoptosis in human virus-transformed and cancer cells compared with normal cells. To overcome the reduction of aralin biosynthesis according to the development of leaves, we established callus tissues from the young shoots. The N terminal amino acid sequence and immunochemical analyses revealed that ca-aralin, an aralin synthesized in the callus, was structurally identical with aralin except for the glycosylation of the B chain. The selective cytotoxic index (ratio of IC50 value against human lung normal cells versus virus-transformed cells) of ca-aralin was 2-fold higher than that of aralin. Thus, these results suggest that ca-aralin is more useful than aralin as an anticancer agent. PMID- 15124097 TI - Inhibitory activity of Juniperus communis on 12(S)-HETE production in human platelets. AB - Extracts of Juniperus communis L. (Cupressaceae) have been evaluated for their inhibitory activity on human platelet-type 12(S)-lipoxygenase [12(S)-LOX]. The methylene chloride extracts of Juniperi lignum, Juniperi pseudo-fructus and the ethyl acetate extract of Juniperi pseudo-fructus showed a significant inhibition on the production of 12(S)-HETE [12(S)-hydroxy-5,8,10,14-eicosatetraenoic acid] at 100 microg/mL (54.0 +/- 6.73, 66.2 +/- 4.03 and 76.2 +/- 3.36%, respectively). From the methylene chloride extract of the wood, cryptojaponol and beta sitosterol were isolated as compounds with inhibitory activity (inhibition at 100 microg/mL = 55.4 +/- 2.80% [IC50 = 257.5 microM] and 25.0 +/- 2.15%, respectively). In addition, a lipid fraction containing unsaturated fatty acids contributed to the in vitro activity of the crude extract. PMID- 15124098 TI - Deoxypodophyllotoxin, a naturally occurring lignan, inhibits the passive cutaneous anaphylaxis reaction. AB - This study examined the effect of a podophyllotoxin derivative, deoxypodophyllotoxin (anthricin), which is a medicinal herb product isolated from Anthriscus sylvestris Hoffm. Deoxypodophyllotoxin was tested in a rat PCA (passive cutaneous anaphylaxis) assay by administering deoxypodophyllotoxin intraperitoneally (1.0 to 10 mg/kg, i.p.) and intravenously (0.25 to 1.0 mg/kg, i.v.). Deoxypodophyllotoxin dose-dependently inhibited the PCA reaction activated by anti-dinitrophenyl (DNP) IgE. The PCA inhibitory activity of deoxypodophyllotoxin was stronger than those of prednisolone and indomethacin, which were used as positive controls. These results suggest that deoxypodophyllotoxin may be beneficial in regulating the immediate-type allergic reaction. PMID- 15124099 TI - The hemolytic activity of epoxydammarane triterpenoids. AB - The hemolytic activity of the epoxydammarane triterpenoids isolated from the Far East species of the genus Betula and their semi-synthetic derivatives was investigated. Comparative studies of epoxydammarane triterpenoid activities at pH 5.5 and 5.0 and at 37 and 42 degrees C showed that 3-oxo, 3,11-dioxo, 3- and 11 acetoxy, as well as 3,11-diacetoxy derivatives had hemolytic potencies lower than the corresponding polyols; triterpenoids with a 3alpha-OH group were stronger than their analogues with a 3beta-OH group; epoxydammaranetriols were somewhat more potent than epoxydammaranediols. Triterpenoids esterified with malonic acid at C-3 possessed the strongest hemolytic activity among the tested compounds. PMID- 15124100 TI - Eremophilane sesquiterpene lactones from Ligularia virgaurea ssp. oligocephala. AB - From an extract of the whole plant of Ligularia virgaurea ssp. oligocephala, three new eremophilane sesquiterpene lactones, 10alpha-hydroxy-1-oxoeremophila 7(11),8(9)-dien-12,8-olide (1), 6alpha,10alpha-dihydroxy-1-oxoeremophila 7(11),8(9)-dien-12,8-olide (2), 6beta,10beta-dihydroxyeremophila-7(11),8(9)-dien 12,8-olide (3), as well as four known sesquiterpene lactones with the same carbon skeleton, named toluccanolides A-C (4, 5, 7) and 6beta-hydroxy-8alpha methoxyeremophila-1(10),7(11)-dien-12,8beta-olide (6) were isolated. Their structures were elucidated by spectroscopic methods including intensive 2D NMR techniques (gCOSY, gHMQC, gHMBC and 1H-1H NOESY for 2) and HR-ESI-MS. PMID- 15124101 TI - Bias toward the null hypothesis in model-free linkage analysis is highly dependent on the test statistic used. AB - Recently, it has been suggested that traditional nonparametric multipoint-linkage procedures can show a "bias" toward the null hypothesis of no effect when there is incomplete information about allele sharing at genotyped marker loci (or at positions in between marker loci). Here, I investigate the extent of this bias for a variety of test statistics commonly used in qualitative- ("affecteds only") and quantitative-trait linkage analysis. Through simulation and analytical derivation, I show that many of the test statistics available in standard linkage analysis packages (such as Genehunter, Merlin, and Allegro) are, in fact, not affected by this bias problem. A few test statistics--most notably the nonparametric linkage statistic and, to a lesser extent, the Aspex-MLS and Haseman-Elston statistics--are affected by the bias. Variance-components procedures, although unbiased, can show inflation or deflation of the test statistic attributable to the inclusion of pairs with incomplete identity-by descent information. Results obtained--for instance, in genome scans--using these methods might therefore be worth revisiting to see if greater power can be obtained by use of an alternative statistic or by eliminating or downweighting uninformative relative pairs. PMID- 15124102 TI - Mutations of the ephrin-B1 gene cause craniofrontonasal syndrome. AB - Craniofrontonasal syndrome (CFNS) is an X-linked craniofacial disorder with an unusual manifestation pattern, in which affected females show multiple skeletal malformations, whereas the genetic defect causes no or only mild abnormalities in male carriers. Recently, we have mapped a gene for CFNS in the pericentromeric region of the X chromosome that contains the EFNB1 gene, which encodes the ephrin B1 ligand for Eph receptors. Since Efnb1 mutant mice display a spectrum of malformations and an unusual inheritance reminiscent of CFNS, we analyzed the EFNB1 gene in three families with CFNS. In one family, a deletion of exons 2-5 was identified in an obligate carrier male, his mildly affected brother, and in the affected females. In the two other families, missense mutations in EFNB1 were detected that lead to amino acid exchanges P54L and T111I. Both mutations are located in multimerization and receptor-interaction motifs found within the ephrin-B1 extracellular domain. In all cases, mutations were found consistently in obligate male carriers, clinically affected males, and affected heterozygous females. We conclude that mutations in EFNB1 cause CFNS. PMID- 15124103 TI - The novel gene encoding a putative transmembrane protein is mutated in gnathodiaphyseal dysplasia (GDD). AB - Gnathodiaphyseal dysplasia (GDD) is a rare skeletal syndrome characterized by bone fragility, sclerosis of tubular bones, and cemento-osseous lesions of the jawbone. By linkage analysis of a large Japanese family with GDD, we previously mapped the GDD locus to chromosome 11p14.3-15.1. In the critical region determined by recombination mapping, we identified a novel gene (GDD1) that encodes a 913-amino-acid protein containing eight putative transmembrane-spanning domains. Two missense mutations (C356R and C356G) of GDD1 were identified in the two families with GDD (the original Japanese family and a new African American family), and both missense mutations occur at the cysteine residue at amino acid 356, which is evolutionarily conserved among human, mouse, zebrafish, fruit fly, and mosquito. Cellular localization to the endoplasmic reticulum suggests a role for GDD1 in the regulation of intracellular calcium homeostasis. PMID- 15124107 TI - [Activation of gene transcription]. PMID- 15124108 TI - [Activation of G-proteins in three dimensions: one step toward the therapeutic inhibition of nucleotide exchange factors]. PMID- 15124109 TI - [High frequency stimulation of the subthalamic nucleus in Parkinson's disease]. PMID- 15124110 TI - [MicroRNA are everywhere]. PMID- 15124111 TI - [Clinical aspects of acute coronary syndromes]. AB - There are two types of acute coronary syndromes : those with or without ST segment elevation. The former require urgent therapeutic measures to reopen the culprit artery (intravenous thrombolysis or primary percutaneous coronary intervention). For the latter, risk stratification is essential and is based upon clinical and biochemical markers. Among them, recent and repeated anginal attacks, ST-segment modifications on admission electrocardiogram, and increased markers of myonecrosis (particularly increased troponin levels) are strong predictors of untoward outcome. According to the risk profile, the initial management is based upon an invasive strategy with powerful antithrombotic medications and urgent angiography, or upon a non-invasive strategy using stress testing, preferably coupled with myocardial imaging techniques. In all instances, secondary prevention measures are determinant to try and stop the progression of the atherosclerotic disease. PMID- 15124112 TI - [Renin-angiotensin system and vascular remodelling]. AB - The renin-angiotensin system (RAS) is compartmented between circulating blood and tissue pericellular space. Whereas renin and its substrate diffuse easily from one compartment to another, the angiotensin peptides act in the compartment where there are generated: blood or pericellular space. Renin is trapped in tissues by low and high affinity receptors. In the target cells, angiotensin II/AT1 receptor interaction generates different signals including an immediate functional calcium dependent response, secondary hypertrophy and a late proinflammatory and procoagulant response. These late pathological effects are mediated by NADPH oxydase-generated free oxygen radicals and NFkappaB activation. In vivo, the tissue binding of renin and the induction of converting enzyme are the main determinants of the involvement of the RAS in vascular remodeling. The target cells of interstitial angiotensin II are mainly the vascular smooth muscle cells and fibroblasts, whereas the endothelial cells and circulating leukocytes are the main targets of circulating angiotensin II. In vivo, angiotensin II participates in the vascular wall hypertrophy associated with hypertension. In diabetes, as in other localized fibrotic cardiovascular diseases, the tissue effects of angiotensin II are mainly dependent on its ability to induce TGF-beta expression. In experimental atherosclerosis, angiotensin II infusion induces aneurysm formation mediated by activation of circulating leucocytes. In these models, the administration of angiotensin II antagonists has beneficial effects on pathological remodeling. Such beneficial effects of angiotensin II antagonists in localized pathological remodeling have not yet been demonstrated in humans. PMID- 15124113 TI - [Polarization of eggs and embryos: some common principles]. AB - Embryonic development depends on the establishment of polarities which define the axial characteristics of the body. In a small number of cases such as the embryo of the fly drosophila, developmental axes are established well before fertilization while in other organisms such as the nematode worm C. elegans these axes are set up only after fertilization. In most organisms the egg posesses a primary (A-V, Animal-Vegetal) axis acquired during oogenesis which participates in the establishment of the embryonic axes. Such is the case for the eggs of ascidians or the frog Xenopus whose AV axes are remodelled by sperm entry to yield the embryonic axes. Embryos of different species thus acquire an anterior end and a posterior end (Antero-Posterior, A-P axis), dorsal and ventral sides (D V axis) and then a left and a right side. PMID- 15124114 TI - [Genetic basis of planar polarity]. AB - Several epitheliums exhibit a clear polarity that lies within the plane of the epithelium. This polarity, referred to as planar polarity or tissue polarity, is oriented perpendicular to the apical-basal polarity of the epithelium. Over the last two decades, the genetic and molecular bases of planar polarity have been intensively investigated in Drosophila. Recent studies have shown that establishment of planar polarity relies on the unipolar distribution of a small number of signaling molecules localizing at the apical cortex. Unipolar localization of planar polarity proteins defines two opposite and complementary cortical domains. These domains show a stereotyped orientation at the tissue level. Positioning of these cortical domains is coordinated at the tissue level by a second class of signaling molecules that form an activity gradient across the epithelium. Together these data have led to a general model of planar polarity establishment. Considering that planar polarity genes have been conserved from flies to vertebrates, this model may be useful for our understanding of epithelium biology in mammals. PMID- 15124115 TI - [Helicobacter pylori and gastric cancer]. AB - Gastric cancer is a worldwide cancer especially frequent in Japan and South America. This cancer affects 10 to 70 people per 100,000 according to the countries. Since the end of the Second World War, the incidence of gastric cancer has been decreasing in France and accounts for less than 10 % of mortality. Helicobacter pylori infection, host genetic background, food regimen are known to be involved in this cancer. Helicobacter pylori should be eradicated in selected patients, such as patients' relatives with documented gastric cancer as well as patients having another gastrointestinal cancer. PMID- 15124116 TI - [Anchoring proteins and cardiac sudden death: how and why?]. AB - Mutations in proteins responsible for ion transport in cardiac tissue can induce a destabilization of electrical function and provoke cardiac sudden death. Identification of a genetic anomaly in a French family that developed the syndrome of cardiac sudden death has revealed a crucial new element in normal cardiac electrical function : Ion channels need to be anchored to specific domains at the plasma membrane by an anchoring protein called ankyrin-B. PMID- 15124117 TI - [Utrophin, a way to cure Duchenne muscle dystrophy]. AB - Duchenne muscle dystrophy results from the absence of dystrophin, a cytoskeletal protein of the muscle fibre. Dystrophin plays an essential role in the integrity of the membrane-associated protein complexes connected to the extracellular matrix. On chromosome 6 is located the gene of a protein presenting 80 % homology with dystrophin : utrophin, which is expressed at the neuromuscular junction. The review examines if utrophin can replace dystrophin and correct the structural and functional characteristics of the myopathy, and how the improvements can be quantitatively expressed. In transgenic mice, deficient in dystrophin, but overexpressing large quantities of utrophin, the latter is found on structures where dystrophin is normally located, histological signs of necrosis disappear and the recovery of functional disorders, specially affecting the mechanical properties of the muscle fibres, can be complete. The review examines also several ways of obtaining overexpression of utrophin in adult mdx mice, such as conditioned expression of the utrophin transgene (using a tetracycline-sensitive transactivator), transfection with viral vectors containing the utrophin cDNA (complete or truncated), actions on factor(s) controlling utrophin expression at the neuromuscular junction (heregulin, 4 N-acetylgalactosamine), and pharmacological ways of inducing expression (NO, arginine). Though partial improvements of the myopathy status have been obtained by these various approaches, they remain limited by their localized action and/or by the moderate level of utrophin expression obtained. Further researchs to overcome these limitations are urgently needed in order to transform the very promising effect of utrophin overexpression into a real treatment of Duchenne myopathy. PMID- 15124118 TI - [Evolution and virulence of primate lentiviruses]. AB - While the AIDS epidemic caused by human immunodeficiency viruses (HIV) has resulted in the death of over 20 million people worldwide, simian immunodeficiency virus (SIV) infection, found in numerous African primate species, does not induce disease symptoms. The factors accounting for this difference between humans and natural host of SIV remain poorly understood. The entangled nature of the host/virus relationship could be the answer, rather than independent virus or host factors. Such a relationship is as a consequence of host/virus adaptation which has evolved over long periods in naturally infected primate species. PMID- 15124119 TI - [Nasopharyngeal carcinomas and Epstein-Barr virus: from epidemiology and detection to therapy]. AB - Nasopharyngeal carcinomas (NPC) challenge clinicians and biologists in various fields including epidemiology, genetics, virology and immunology. These tumours have a striking geographical distribution. They are constantly associated with the Epstein-Barr virus (EBV) and contain a massive lymphocytic infiltrate. Their study has major implications especially at this moment while a pathological role of EBV is suspected in several other human epithelial malignancies (for example gastric, mammary and thyroid carcinomas). The North-South Workshop on Nasopharyngeal Carcinoma was held at the Institut Gustave-Roussy in early December 2003. Its main goal was to support the exchanges between clinical research on NPC in the South and basic research in the North. With regard to epidemiology and genetics, the main information was the possible existence of several susceptibility genes (including two of them on the 4p and 5p chromosomes). In virology, participants have emphasized the selection of peculiar EBV variants within the malignant cells and the expression of novel oncogenic viral proteins : LMP2 and BARF1. Cellular gene alterations also contribute to NPC development, especially inactivation of tumor suppressor genes located on the 3p chromosome. Therapeutic research was not forgotten. Hope of higher rate of cure relies on improved ballistic processes in radiotherapy (IMRT) and on the development of targeted therapeutics : induction of the lytic/productive viral cycle, gene therapy with conditional replicative adenoviruses, antitumor vaccination directed against the viral protein LMP2. PMID- 15124120 TI - [Studies of several analytical methods for antioxidant potential evaluation in food]. AB - Oxidation control is necessary to manage the evolution of complex biological system, particularly in food whose degradation could have consequences on food security. After description of context and oxidation mechanisms, several analytical methods to evaluate the additive antioxidant potential are presented. This evaluation is performed either by quantification of products (in particular hydroperoxydes) using direct or indirect photometric techniques and chemical titration with suitable reactants or on the effectiveness to trap free radicals with modelized systems that can generate them. Methods based on the comparison of radical trapping ability between an additive and Trolox (particularly Trolox(R) equivalent antioxydant capacity, TEAC) can be applied to many products whatever the hydrophily or the hydrophobia of the medium. PMID- 15124121 TI - [From the conception of the PRINS to its coronation]. AB - As a non-isotopic molecular cytogenetic technique, the primed in situ (PRINS) labelling reaction represents a major technological progress achieved in the past decade. It has become a routine technique for the microscopic visualization of specific DNA sequences in cells and nuclei and constitutes a good alternative to the fluorescence in situ hybridization (FISH) procedure. Among the multiple advantages that characterize the PRINS technique, specificity, rapidity, reliability, reproducibility, and cost-effectiveness can be mentioned. PRINS can be in addition associated with other techniques like FISH, indirect immunofluorescence, and nick translation. The most recent developments show the great potential of this technique. Now PRINS can be used to study single-copy genes and, consequently, can be routinely used to investigate deletions associated with microdeletion syndromes. Therefore, the PRINS technique has the potential to become a widely used molecular cytogenetic tool in clinics and research. This short review presents how the PRINS technique contributed to further the understanding of biological phenomena and describes the different possibilities and applications of the PRINS method in several biological and clinical fields (pre-implantation testing, prenatal, constitutional and oncologic genetic diagnosis). PMID- 15124122 TI - [Collective of users in the fields of AIDS and drug addiction]. AB - The extent of mobilization caused by the epidemy of AIDS was underlined many times. Because of a logic of selective dissemination, infection by HIV not only started action of patients and their close relations, but also that of the two most affected social groups : homosexual men and drug users. Existing since almost a century, the collectives of patients have shown various configurations, from consensual groups which developped in the thirties to protestor groups emerging within the seventies. We show here how both in AIDS and drug addiction, the collectives of users multiplied through differentiated public identification choices. In the fight against AIDS, all the forms of mobilization coexist. Concerning drugs usage, actions are less diversified and divide in two main categories: groups of interest and self-help groups. PMID- 15124123 TI - [Analysis of gene expression in the human nephron]. PMID- 15124124 TI - [Budget. 1848]. PMID- 15124125 TI - [A squall about DNA chips]. PMID- 15124126 TI - Proceedings from the 3rd Annual Updates in Hematology and Oncology. July 2003, Sea Island, Georgia, USA. PMID- 15124127 TI - Novel agents and incremental advances in the treatment of head and neck cancer. AB - Cancer of the head and neck is an important medical problem, with approximately 46,500 cases predicted in the United States alone in 2003. Worldwide, more than 600,000 cases are anticipated. While several different histologic subtypes of head and neck cancer are seen in different parts of the world, more than 90% of tumors diagnosed in the United States are squamous cell carcinomas. Major strides in the management of this disease have been made in the last decade. These include, but are not limited to, the evolution of organ preservation, the increasingly well recognized role of concurrent chemoradiation therapy as either definitive therapy for unresectable disease or adjuvant therapy for high-risk surgical disease, and significant improvements in cytotoxic chemotherapy. The role of chemotherapy in this disease has been a subject of debate. Chemotherapy is now routinely included in the multimodality treatment of unresectable disease of the oral pharynx, larynx, and oral cavity. There is now increasing evidence supporting the role of induction chemotherapy in head and neck cancer. As intensified chemotherapy and radiation therapy have improved local control, the increasing incidence of distant metastases has necessitated the need for enhanced systemic control. These approaches are the topics of extensive investigations. PMID- 15124128 TI - The care of the lung cancer patient in the 21st century: a new age. AB - Lung cancer is by far the leading cause of cancer-related death within the United States and throughout the world, with a global incidence estimating 1,240,000 in 2001. While median survival has improved from 7 months in 1960 to 16.8 months in 2001 for all patients diagnosed with lung cancer, much of this progress relates to improved surgical techniques, combined modality therapy for locally advanced disease, improved symptom palliation, and moderate but real improvements in survival of stage IV disease. However, much work remains to be done in a field in which only 15% to 17% of patients live 5 years. The role of chemotherapy in the last decade has expanded substantially, with evidence for extension of median survival in stage IV from 4 months to 8 to 10 months, improvements in symptom control, as well as 1-year survival almost tripling from 11% to 14% to 32% to 37% with modern chemotherapy regimens. Furthermore, progress has also been seen in the use of concomitant chemoradiotherapy, which has become the mainstream approach for treating patients with locally advanced non-small cell lung cancer. Finally, trials with cisplatin-based chemotherapy suggest that for resectable non small cell lung cancer, improvement in disease-free and overall survival at 5 years is in the range of 4% to 5%. With the development of multiple small molecules and monoclonal antibodies targeting important growth factor receptors, oncogenes and tumor-suppressor genes known to be aberrant in lung cancer, there is hope for further incremental improvements in the treatment of this deadly disease. PMID- 15124129 TI - Update on the management of connective tissue malignancies. AB - Approximately 11,000 new cases of connective tissue malignancies are anticipated in 2004. These diseases can be divided into soft-tissue sarcomas, sarcomas of bone, and gastrointestinal stromal tumors. Optimal management of these diseases requires a multidisciplinary team with expertise in surgery, pathology, radiotherapy, and chemotherapy. Over half of patients with stage III soft tissue and bone sarcomas are cured, as are some patients with metastatic disease. Imatinib mesylate has been an important advance in the treatment of gastrointestinal stromal tumors. PMID- 15124130 TI - New targeted therapies in breast cancer. AB - Targeted therapy for breast cancer has existed since 1986, when Beatson published his observations on oophorectomy. In the past 5 years monoclonal antibodies and tyrosine kinase inhibitors have been evaluated in phase I and II clinical trials of breast cancer. Trastuzumab (Herceptin; Genentech, Inc, South San Francisco, CA) was the first such agent to be approved by the US Food And Drug Administration, following findings that it improved survival in patients with metastatic breast cancer. Gefitinib (ZD1839, Iressa; AstraZeneca, Wilmington, DE) has little activity as a single agent in unselected patients, but, preclinical data have suggested synergy with tamoxifen and other hormonal agents, as well as other growth factor inhibitors. Bevacizumab (Avastin; Genentech, Inc), a monoclonal antibody against vascular endothelial growth factor is being evaluated in metastatic breast cancer. Furthermore, agents targeting other pathways, such as the Ras pathway with farnesyl transferase inhibitors, and mTOR with rapamycin analogues are currently under investigation. In the next few years, and as trials with the above agents mature, we will further define the role of these targeted agents in the treatment of breast cancer. PMID- 15124131 TI - Novel therapeutic strategies for androgen-independent prostate cancer: an update. AB - Close to 30,000 men will die from prostate cancer in the United States in 2003. Hormonal ablation, the basis of systemic therapy, will invariably fail to control the progression of metastatic prostate cancer in the long run. For many years the only available therapeutic modalities for patients with metastatic androgen independent prostate cancer have been second-line hormonal maneuvers with estrogens or steroids and chemotherapy. So far, chemotherapy has been shown to confer adequate palliation but no overall survival benefit in prospective randomized controlled trials, and the only chemotherapy drugs approved by the US Food and Drug Administration are mitoxantrone and estramustine. Novel treatment strategies that aim at specific signaling pathways, apoptosis, differentiation, or specific membranous targets are being developed. Such new therapeutic modalities, along with recent data on immunotherapy and bone targeting strategies, will be reviewed here. PMID- 15124132 TI - Update on the systemic treatment of malignant melanoma. AB - The purpose of this article is to review some of the recent advances and disappointments in the systemic treatment of melanoma and to highlight some of the ongoing trials in this area. Two major advances in the staging of melanoma are the new American Joint Committee on Cancer staging system and the use of the sentinel node biopsy. Interferon remains the standard adjuvant therapy for high risk patients. Ongoing trials are evaluating 1 month of high-dose interferon versus observation in intermediate-risk patients and comparing standard interferon with biochemotherapy. Allogeneic and peptide vaccines and granulocyte macrophage colony-stimulating factor are also being evaluated. Dacarbazine and high-dose IL-2 are the only US Food and Drug Administration approved systemic treatments for stage IV disease. Several new agents are being evaluated. Melanoma remains a prototype disease in which patients should be encouraged to participate in clinical trials. PMID- 15124133 TI - Emerging molecular therapies for brain tumors. AB - This article briefly reviews the basic research findings on brain tumors made in the last decade and how they are now impacting clinical trials. Major improvements in our understanding of the genetic, molecular, and biological mechanisms that lead to brain tumor development have been made. These include mechanistic insights on how tumor cells overcome cell cycle control, evade programmed cell death, induce blood vessel formation, and escape immune regulation. These advances have been tremendously accelerated by the availability of novel whole-genome analysis technologies. These findings are now being translated into innovative clinical trials that provide new hope for patients with these devastating diseases. PMID- 15124134 TI - Cancer screening. AB - Cancer screening is a complicated science. Each screening intervention must be carefully assessed before it is widely implemented. A screening test can falsely appear useful as it finds disease at an early stage and leads to intervention and cure. Such a test can be harmful to the population screened if it commonly finds disease that fulfills the pathologic criteria of cancer but behaves indolently (meaning it would never harm the host). Such "pseudo-disease" or "overdiagnosed disease" has been demonstrated in many malignancies including cancers of the lung, breast, and especially the prostate. The nature of each specific screening test and each disease is such that some screened patients may receive unnecessary treatment with all its complications and risk. Alternatively, some screening technologies have been proven useful providing net benefit to the population screened. Often these beneficial technologies are underused. These screening technologies if widely implemented have the potential of saving countless lives. Many available screening tests have tremendous potential in terms of benefit, but have yet to be fully assessed. At the minimum, patients should be informed of what is known, what is not known, and what is believed about these tests. PMID- 15124135 TI - An update on the management of relapsed and primary refractory Hodgkin's disease. AB - With the use of combined modality therapy for early stage disease and a risk adapted approach for advanced stage disease, nearly 90% of patients with Hodgkin's lymphoma are cured with initial therapy. However, in patients who have primary refractory or relapsed disease, high-dose therapy and autologous stem cell transplantation is the best curative option. The use of peripheral blood progenitor cells has decreased transplant related mortality to less than 3%; but long-term progression-free survival as increased minimally. Although prognostic factors have been used to tailor therapy in de novo Hodgkin's lymphoma their utility in the relapsed-refractory setting has not been exploited. This update will discuss these important prognostic factors and try to guide oncologists in treatment decisions in this setting. PMID- 15124136 TI - Treatment of chronic lymphocytic leukemia. AB - Treatment options for patients with chronic lymphocytic leukemia have changed over the past two decades. This article reviews the experience accumulated with the use of alkylating agents alone and in combination; purine analogues alone and in combination and monoclonal antibodies such as rituximab, and alemtuzumab alone and in combination. The results obtained with different treatment strategies are summarized, compared, and reviewed. PMID- 15124137 TI - Advances in follicular lymphoma. AB - Follicular lymphoma is a usually indolent lymphoma that responds well to chemotherapy. While multiple treatments show a good response rate, most patients relapse. Emerging therapies, such as antibody therapy and stem cell transplantation, are increasingly being used to try to lengthen response time. This article will review the available treatments for follicular lymphoma and discuss the studies supporting newer strategies of treatment. PMID- 15124138 TI - Update on myelodysplastic syndromes: new approaches to classification and therapy. AB - Myelodysplastic syndromes have long provided hematologists with difficult therapeutic challenges, and until recently treatment options beyond supportive care were limited. Recent advances in our understanding of hematopoiesis, immunology, and genetics have led to a better understanding of the natural history of these disorders and have facilitated development of more rational and targeted treatment approaches. A number of promising agents are in various phases of study, including arsenic trioxide, CC5013, the farnesyltransferase inhibitors, and DNA methyltransferase inhibitors. In addition, less intensive strategies for allogeneic stem cell transplantation now permit us to offer potentially curative therapy to a larger proportion of patients. Optimal management of an individual patient requires consideration of the disease and its expected course, available treatment options, the patient's age and condition, and an ongoing assessment of the goals of therapy. PMID- 15124139 TI - Update in acute leukemia 2003: a risk adapted approach to acute myeloblastic leukemia in adults. AB - Acute myeloblastic leukemia represents a heterogeneous group of diseases. The diagnosis and prognosis is most accurately provided by pretreatment assessment of the clonal molecular genetic derangement responsible for the disease, often provided by cytogenetic analysis. Other prognostic features include patient age, antecedent myelodysplasia, prior chemotherapy, and the presence of FLT-3 mutations. Accurate assessment of prognosis permits a risk-adapted treatment approach to maximize probability of cure and minimize treatment-related toxicity. The majority of patients with promyelocytic leukemia with the PML/RARalpha fusion gene can be cured with an all-trans-retinoic acid and anthracycline-based treatment program. All other patients are typically given cytarabine and anthracycline-containing induction regimens, although some with particularly poor prognosis disease may be more appropriate candidates for experimental induction therapies. Postinduction treatments include further conventional chemotherapy, stem cell transplant strategies, and experimental approaches. Issues pertinent in selecting treatments for patients in the different risk categories are reviewed. PMID- 15124140 TI - Cellular immunotherapy and cancer. AB - Recent data indicate that cellular immune responses can produce an anticancer effect. Two types of cellular immune responses appear to be clinically relevant: antigen specific immunity mediated by T cells that recognize tumor-associated peptide antigen expressed on surface HLA class I or class II molecules, and antigen nonspecific immune responses mediated by natural killer (NK) cells that are activated by the failure to recognize cognate "self" HLA class I molecules. The graft-versus-tumor effect of allogeneic transplants using HLA-matched donors is mediated by antigen-specific T cells, while allogeneic transplantation using HLA mismatched donors can also lead to donor NK cells with potent antitumor activity. HLA mismatched haplo-identical transplants can exert a powerful antileukemia effect based on expansion of antigen nonspecific donor NK cells, but slow recovery of donor T cells is associated with a profound immune deficiency and increased opportunistic infections that limit the overall clinical success of this type of transplant maneuver. The focus for most immunologic maneuvers designed to enhance the graft-versus-tumor effect has been on strategies to expand or enhance the activity of antigen-specific donor T cells that recognize host tumor cells. In the case of hematologic malignancies, the activity of donor T cells against tumor cells is regulated by dendritic cells, which can augment or inhibit cellular immune responses. Future successes in enhancing the patients' own cellular immune responses to cancer will likely be based on activation of T cells by the appropriate dendritic cell subset and, in the case of tumors that have down-regulated HLA class I expression, by activated NK cells. PMID- 15124141 TI - New options for the treatment of invasive fungal infections. AB - Fungal infections continue to plague patients undergoing treatment of malignancies. In fact, the incidence as well as the types of fungal species causing serious invasive fungal infections is increasing. Candida infections have become the fourth leading cause of nosocomial blood stream infections. Moreover, in addition to Aspergillus, non-Aspergillus species have emerged as etiologic agents of mold infections in cancer patients. Recently, there have been new and much needed re-enforcements to the antifungal armamentarium. These include the broader triazole antifungal agents that are in clinical trials (posaconazole) or have been recently approved (voriconazole) and the echinocandins, a new antifungal class of which the first approved agent caspofungin is now available. We review the clinical data, side effects, drug interactions, and unique spectrum of activity of these newer antifungal agents against both Candida and clinically relevant mold species. PMID- 15124142 TI - Multiple myeloma: the role of transplant and novel treatment strategies. AB - Multiple myeloma will be diagnosed in over 14,000 new patients over the next year. While myeloma is rarely curable, recent advances have improved the care and outlook for most patients. Recent progress has been made on the molecular characterization of myeloma, cytogenetics, prognosis, and supportive care. Randomized trials have proven the efficacy of high-dose therapy supported by autologous transplantation. Reduced-intensity allogeneic transplantation, associated with much less morbidity and mortality than standard myeloablative transplantation, can now be offered to more myeloma patients. And finally, a greater understanding of the pathophysiology of myeloma development and progression, growth, and survival have led to the development and clinical use of several novel therapeutics. This review will discuss recent improvements in therapies for myeloma focusing on transplantation and novel targeted therapies. PMID- 15124145 TI - Ethical use of long-acting medications in the treatment of severe and persistent mental illnesses. AB - volunteerism Mental illnesses are prevalent, cause great suffering, and are burdensome to society. Traditional "depot" antipsychotic agents are used to treat the most severely and persistently mentally ill individuals. They will soon be joined by new atypical antipsychotic medications in long-acting formulations. These long-acting medications pose special ethical issues, but may greatly benefit some people who suffer from severe and persistent mental illnesses. The objective of the current report is to highlight ethical considerations related to the use of long-acting antipsychotic medications. An analysis centering on the ethical concepts of voluntarism, beneficence, and justice is performed, integrating relevant empirical evidence and bioethics principles. Two main conceptual issues related to constraints upon voluntarism and coercion exist. Careful examination of ethically important empirical evidence suggests that voluntarism may not be perceived as an issue for some persons receiving depot antipsychotic medications. A favorable balance of benefits and risks has been documented for some individuals with severe and persistent mental illnesses. Access to care, innovative treatments, and long-acting antipsychotic medications may arise as an issue of justice and nondiscrimination in the care of mental illness. Considerations of justice indicate that long-acting medications may need to be used more frequently and earlier in the course of severe and persistent mental illness for select patients. We conclude that great care should be given to ethically important issues surrounding voluntarism, beneficent care, and equitable access to innovative psychiatric treatments, especially for persons who carry the burden of stigma as well as severe and persistent mental illness. PMID- 15124146 TI - Affective disorders in the first-degree relatives of bipolar probands: results from the South Island Bipolar Study. AB - The current study was performed to document observed rates of affective disorders in the first degree relatives of probands with bipolar I or II disorder; to determine whether bipolar II probands have an excess of bipolar II relatives; and to determine whether bipolar probands with a history of one or more suicide attempts have more relatives who have also made suicide attempts. Bipolar probands with positive family histories of affective disorder were recruited from a variety of sources for a study on the molecular genetics of bipolar disorder. Probands and relatives were interviewed with the Diagnostic Interview for Genetic Studies (DIGS) and blood was obtained for DNA extraction and genetic analyses. Among 423 first-degree adult relatives of 153 bipolar probands, 7% (29) had bipolar I disorder, 7% had bipolar II disorder, and 7% had bipolar not otherwise specified (NOS) disorder, making 21% of relatives with any bipolar disorder. A further 42% of relatives had a depressive disorder and only 38% had no affective disorder. A suicide attempt by a proband was not associated with any increase in suicide attempts by relatives. We conclude that while unipolar depressive disorders are the most common affective disorders in the first-degree relatives of bipolar probands, extension of the bipolar phenotype to include bipolar spectrum disorders results in 21% of relatives having any bipolar disorder. PMID- 15124147 TI - Screening and diagnostic utility of self-report attention deficit hyperactivity disorder scales in adults. AB - Attention deficit hyperactivity disorder (ADHD) in adults is associated with significant social, legal, occupational, and psychiatric difficulties. The estimated prevalence of ADHD in the adult population is between 0.3% and 5%. Recent interest in the condition in adults has been accompanied by the appearance of a number of readily available scales for screening adults and aiding in the diagnosis of ADHD in this age group. However, there are few published data on the validity and reliability of such measures. We examined the diagnostic and screening utility of three ADHD scales (Adult Rating Scale [ARS], Attention Deficit Scales for Adults [ADSA], and Symptom Inventory for ADHD) in 82 adults presenting for ADHD evaluation. All three instruments were sensitive to the presence of symptoms in adults with ADHD (correctly identifying 78% to 92% of patients with ADHD), but a high proportion of individuals with non-ADHD diagnoses screened positive (incorrectly identifying between 36% and 67% of non-ADHD patients). Our results suggest that the use of such measures for screening and as an aid in diagnosis should be approached with considerable caution. PMID- 15124148 TI - Asperger's disorder: a review of its diagnosis and treatment. AB - Asperger's disorder is a pervasive developmental disorder that shares similar features of social impairment disorder, restricted interests, and repetitive behaviors with autistic disorder. Although both Asperger's disorder and autistic disorder persist into adulthood, autistic disorder is usually apparent before the age of 3, while Asperger's disorder usually manifests itself at preschool age. Asperger's disorder in the majority of cases is not associated with delay in language development and there is an increased likelihood to seek social interactions and to engage in activities and friendship with others. In contrast to autistic disorder, most Asperger's disorder patients have normal intellectual functioning and some have motor clumsiness. Although the etiology of Asperger's disorder is still undetermined, this article will review the assessment and treatment interventions that could improve the prognosis of this illness. The historical background, epidemiology, diagnostic features, differential diagnosis, and course and overall management/treatment of Asperger's disorder will be discussed. Despite the absence of a cure for Asperger's disorder, the awareness of its distinctive clinical features that differentiate it from autistic disorder could improve its prognosis and differentiate response to treatment and comorbid conditions. PMID- 15124149 TI - Life events and personality factors in children and adolescents with obsessive compulsive disorder and other anxiety disorders. AB - The association among life events, personality factors, and anxiety disorders in children and adolescents was assessed in 28 children with obsessive-compulsive disorder (OCD), 28 children with other anxiety disorders (AD), and 24 normal controls using the Life Events Checklist (LEC) and the Junior Temperament and Character Inventory (JTCI). No significant differences were found among the groups for demographic and clinical characteristics. Children with OCD had significantly more total life events and more negative life events in the year before onset than normal controls, and they perceived the life events as having more impact. Scores for children with other AD fell between the other two groups for most of the life event parameters. The only specific life event that distinguished children with AD from normal controls was major illness or injury of a relative. High anxiety levels and older age--but not depression level- predicted a greater perceived impact of life events. Children with OCD and other AD both scored higher than normal controls on the harm avoidance parameter of the JTCI. Harm avoidance scores correlated positively and significantly with the reported occurrence of negative life events and their perceived impact. Thus, quantity, quality, and specificity of life events may be associated with AD in young people, especially OCD. This association may be related to the personality characteristic of harm avoidance. PMID- 15124150 TI - Outcome of psychiatric treatment: what is relevant for our patients? AB - This study investigated relevant outcome domains in the patient's perspective following psychiatric outpatient treatment for non-psychotic, non-substance related disorders. Questionnaires, including the Client Satisfaction Questionnaire (CSQ) and the Bern Inventory of Treatment Goals (BIT-C) applied as a broad typology of outcome domains, were mailed 1 year after treatment to outpatients who had undergone eight or more therapy sessions. Patients reported a wide range of relevant outcomes, including changes with respect to the interpersonal domain, their self-concept, and existential issues. Changes in depressive and anxiety symptoms were rated as particularly important; the reports of both symptomatic and more integral changes were related to treatment characteristics, patient's diagnostic category, and patient's employment status. Patient satisfaction was particularly related to reported changes in the interpersonal domain. This exploratory study provides evidence that traditional outcome measures that include mood, anxiety, and fear symptoms continue to assess the most important areas for change in patients' views. However, they might miss relevant therapeutic achievements in some of our patients, particularly in those suffering from adjustment and personality disorders. The use of measures that include dimensions such as personal growth, purpose of life, and positive relations with others may record important changes in these patients. PMID- 15124151 TI - Parental rearing style, premorbid personality, mental health, and quality of life in chronic regional pain: A causal analysis. AB - The aim of the current study was to establish the causal model among parental bonding, personality characteristics, mental health, quality of life, and chronic regional pain (CRP). Thirty CRP patients and 56 mental illness patients were compared using the Visual Analogue Scale (VAS), Chinese Health Questionnaire (CHQ), Eysenck Personality Questionnaire (EPQ), World Health Organization Quality of Life-Brief-Tawain Version (WHOQOL-BREF-TW), and Parental Bonding Instrument (PBI). There were significant differences in mental health, personality characteristics, and quality of life between the CRP and mental illness groups. Structural equation modeling showed that parental bonding could directly affect personality characteristics, and, hence, directly impact disease and quality of life. CRP is different from mental illness in many dimensions. In this study, CRP appeared to be caused by actual physical dysfunction rather than mental dysfunction. PMID- 15124152 TI - Patient characteristics and diagnostic discrepancy in first-episode psychosis. AB - Despite the operational criteria in diagnostic systems there is still marked diversity between clinical and research diagnoses in populations with psychotic disorders. The objective of the current study was to explore the association of patient-related factors with diagnostic agreement between clinical diagnoses and Schedules for Clinical Assessment in Neuropsychiatry (SCAN-2) diagnoses in first episode psychosis. The sample included 80 consecutive patients. As explanatory variables we used demographic characteristics (gender, age, living circumstances, education, and social activities), measures of psychopathology (Positive and Negative Symptom scale [PANSS], Hamilton Depression Scale, Global Assessment of Functioning Scale [GAF], and Strauss-Carpenter Scale), duration of untreated psychosis, and diagnostic category according to SCAN-2 interview. The overall agreement value between the clinical and research diagnoses was 0.55 (kappa). In the whole sample low scores on the PANSS negative subscale, low level of education, and high score on the PANSS item for delusions predicted diagnostic discrepancy. Acute and transient psychotic disorder as a research diagnosis predicted diagnostic agreement. In the schizophrenia group, young age and lack of social activities predicted diagnostic agreement. Bivariate comparisons of treatment compliance, perceived medication side effects, or negative attitudes towards treatment showed no associations with diagnostic agreement. The results confirm some of the findings in the few previous studies. The diagnosis of schizophrenia is likely to be delayed and there is a need for further education with clinicians in recognizing the symptoms of schizophrenia. PMID- 15124153 TI - Obsessive-compulsive symptoms in patients with panic disorder. AB - Comorbidity studies have shown an important association between panic disorder (PD) and obsessive-compulsive disorder (OCD). The aim of the current study was to evaluate the prevalence of obsessive-compulsive symptoms (OCS) and OCD in patients with PD. Forty-eight consecutive PD cases (DSM-IV diagnostic criteria) referred to a Brazilian university hospital clinic were studied. The Yale Brown Obsessive Compulsive scale (Y-BOCS) checklist was used to identify the OCS. Subclinical OCD was considered when subjects met all but one DSM-IV criteria for OCD (symptoms did not cause significant distress and interference, did not last more than 1 hour per day, or were not considered excessive or irrational), and OCS when only the criterion for presence of obsessions or compulsions was met. Twenty-nine (60.4%) of the 48 patients evaluated (19 men and 29 women) had at least one OCS: nine (18.8%) had mild OCS, 11 (22.9%) had subclinical OCD, and nine (18.8%) had comorbid OCD. Therefore, 41.7% of the patients had either clinical or subclinical OCD. OCS occurred more frequently in women and, in 70.4% of the cases, preceded the onset of PD. Our results suggest that it is important to evaluate systematically the co-occurrence of OCS in patients with PD, due to the considerable overlap found in symptoms, which may have therapeutic implications. As panic symptoms are usually the main complaint, OCS are often found only when directly investigated. PMID- 15124154 TI - Korean version of the diagnostic interview for genetic studies: Validity and reliability. AB - The Diagnostic Interview for Genetic Studies (DIGS), developed in 1994 by the National Institute of Mental Health (NIMH), was translated into Korean and tested for reliability and diagnostic validity. Concurrent validity was tested using the Structured Clinical Interview for DSM-IV (SCID) and clinical diagnoses in 53 patients, most of whom had either schizophrenia or bipolar disorder. Inter-rater reliability was tested in 24 patients. Test-retest reliability was also tested in 17 patients. Overall and specific diagnostic validity for the Korean version of DIGS (DIGS-K) was excellent for most diagnoses. Inter-rater and test-retest reliability for overall and specific diagnoses also ranged from fair to excellent. For schizoaffective disorder, the test-retest reliability of DIGS-K was in a fair range, although the level was lower than that of other diagnoses. However, its diagnostic validity and inter-rater reliability was below fair range. In conclusion, DIGS-K appears to be a reliable interview for major psychiatric disorders. PMID- 15124155 TI - Resolution of delusional depression after recovery from delirium. AB - Little attention has been given to the effects of delirium on the course of depression. In clinical practice, we sometimes observe delirium brought on incidentally by severe physical illness or therapeutic drugs such as tricyclic antidepressants. Recently, investigators have discussed whether delirium can in fact have a beneficial effect on the course of depression. We present three cases of delusional depression in which depressive symptoms resolved after patients recovered from incidental delirium caused in two cases by medication, and by respiratory distress leading to asphyxiation in the third. We surmise that delirium may create a biological effect similar to that of electroconvulsive therapy (ECT), which is widely hailed as an effective treatment for delusional depression. Retrograde amnesia caused by delirium and the supportive milieu during treatment of the delirium may have a beneficial psychological effect on recovery from delusional depression. PMID- 15124156 TI - Eating behavior and other distracting behaviors while driving among patients with eating disorders. AB - The current study sought to better characterize eating behavior, binge-eating behavior, and other potentially problematic, distracting behaviors while driving in patients with eating disorders. Forty patients with eating disorders who reported eating in their car at least once per week were included. Thirty subjects with eating disorders reported binge-eating while driving. A surprisingly high number of subjects reported engaging in a variety of distracting behaviors, including changing clothes, reading, applying make-up, and combing/brushing their hair while driving. Potentially problematic behaviors such as binge-eating while driving should be inquired about during assessment, and if found should be addressed in treatment planning. PMID- 15124162 TI - Audiovestibular findings in patients with delayed and idiopathic endolymphatic hydrops: a comparative study. AB - OBJECTIVE: The objective of this study was to analyze the neuro-otological findings in patients with delayed endolymphatic hydrops and compare them with audiovestibular abnormalities found in patients with idiopathic endolymphatic hydrops. Study design A retrospective open case study. SETTING: A referral ambulatory dizziness clinic. PATIENTS: Medical records of 30 patients with delayed endolymphatic hydrops and of 33 patients with idiopathic hydrops were analyzed. Outcome measures Clinical, electronystagmographic, caloric, and audiometric results in both groups of patients. RESULTS: No differences in the prevalence of abnormalities on clinical testing, electronystagmography (including rotatory chair), or caloric testing were found between the groups. The clinical outcome of conservative therapy was better in patients with idiopathic endolymphatic hydrops (P =.03). CONCLUSIONS: These findings are in keeping with the presumed common pathophysiology of delayed and idiopathic endolymphatic hydrops. However, it is important to identify patients with delayed hydrops in view of the chance of the potential poor response to medical therapy. PMID- 15124163 TI - Resection of juvenile angiofibroma using the Le Fort I approach. AB - BACKGROUND: Juvenile angiofibroma (JA) is a rare tumor that occurs exclusively in adolescent male patients. The mainstay treatment for JA is complete surgical excision. Acceptable surgical approaches for the resection of JA include transantral, transpalatal, combined, endoscopic, and Le Fort I procedures, among others. Because exposure of the entire extension of the tumor may not possible, the recurrence rates after surgical treatment of JA may be as high as 55%. The purpose of this study was to evaluate the results after using the Le Fort I technique for the resection of JA. METHODS: We retrospectively studied 19 patients with JA submitted to surgical resection by using the Le Fort I approach from March 1983 to September 2002. Data regarding demographic characteristics, tumor topography, use of embolization, recurrence, and complication of treatment were obtained. RESULTS: Patient age at the time of diagnosis ranged from 8 to 26 years with a mean age of 16 years. The most common tumor site was the nasopharynx in 100% of the cases, pterygopalatine fossa (95%), nasal cavity (84%), and sphenoid sinus (63%). Angiography and embolization were performed preoperatively in 57% of the patients. The follow-up period ranged from 1 to 19 years (mean, 9.7 years; median, 8.0 years). One patient experienced malocclusion after surgery. However, we did not observe any recurrences during the follow-up period. CONCLUSION: We conclude that the Le Fort I approach is a safe technique that permits the total resection of the JA with a low rate of postoperative complications and a low rate of recurrence. PMID- 15124164 TI - Sinonasal undifferentiated carcinoma: case series and literature review. AB - PURPOSE: The purpose of this study was to understand the natural history and progression of sinonasal undifferentiated carcinoma (SNUC) to establish optimal management guidelines. Methods and materials We analyzed 8 consecutively treated patients diagnosed with SNUC between 1995 and 2002 at UCLA Medical Center. Staging was classified by the Kadish System with 7 patients presenting at stage C and 1 patient with stage B disease. Five patients received surgery. Four of these 5 patients received adjuvant radiotherapy, with 2 patients receiving it concurrently with chemotherapy (cisplatinum/5-fluorouracil). One patient received surgery alone as definitive management. Of the remaining 3 patients who did not receive surgical treatment, concurrent chemotherapy and radiation was used. RESULTS: At last follow-up, 6 of the 8 patients were still alive (overall survival 75%) with mean survival time of 20.6 months. However, only 2 of the 6 were alive and free of disease (disease-free survival 25%) with a mean disease free survival time of 12.3 months. Locoregional recurrence occurred in 5 of the 8 patients (63%). Time to recurrence ranged from 3 to 30 months. Distant metastasis presented in 4 of the 8 patients (50%), all with disease spread to bone. Two of the 4 patients with distant metastasis were found to have locoregional disease recurrence at the same time. Time to metastasis ranged from 2 to 30 months. Results also show that the 5 patients who received surgery are still alive with a mean survival time of 23 months at last follow-up. Meanwhile, 1 of the 3 patients who received no surgical therapy is alive, with a mean survival time of 16.7 months in this group. CONCLUSIONS: SNUC has proven to have a poor prognosis. Although limited by small numbers, this study along with reported series in the past appear to suggest longer survival results with aggressive multimodality therapy, especially with the incorporation of complete surgical resection. PMID- 15124165 TI - Office-based foreign-body management using videoendoscope. AB - BACKGROUND: The videoendoscope has a small charge-coupled device (CCD) chip built into its tip that provides a clear image. This report concerns office-based pharyngeal and laryngeal foreign-body management using a videoendoscope. METHODS: Three types of videoendoscopes (videoendoscopes equipped with and also without a hood at their tips and a rigid videoendoscope) were used in this study. Seventeen patients who complained of pharyngeal and laryngeal foreign bodies were treated with these videoendoscopes. RESULTS: The advantages of this intervention were (1) videoendoscopes presented clear dynamic color images on a color video monitor and provided excellent resolution and recording and thus yielded high diagnostic accuracy and fine intervention; (2) the videoendoscope's diameter was relatively small and resulted in less discomfort for patients, even for children; (3) patients could be examined and treated in a sitting position on a procedure chair at the otolaryngology outpatient clinic, which obviated general anesthesia; (4) the videoendoscope equipped with a hood enabled treatment in a closed cavity like the hypopharynx; (5) minute foreign bodies located at the portion of the pharynx, which were hard to examine, could be depicted clearly; (6) pernasal endoscopy allowed the doctor to examine patients who had a strong gag reflex; and (7) good image documentation on the color video monitor allowed the physician to carry out safe intervention. A disadvantage of this procedure was that the extraction of different kinds of foreign bodies was limited, but it depended in part on the efficacy of the forceps. CONCLUSION: Foreign-body extraction using a videoendoscope is one of the reliable procedures that has widened the indications for office-based endoscopy. PMID- 15124166 TI - NARES: a risk factor for obstructive sleep apnea? AB - BACKGROUND: Nonallergic rhinitis with eosinophilia syndrome (NARES) constitutes a rare nasal condition characterized by a chronic, eosinophilic inflammation. Patients' major complaints constitute nasal congestion and rhinorrhea. Obstructive sleep apnea syndrome (OSAS) is a potentially life-threatening condition characterized by recurrent episodes of obstruction of the upper airways resulting in oxygen desaturation. Nasal congestion constitutes one predisposing factor for OSAS. OBJECTIVE: The purpose was to study whether NARES constitutes a risk factor for OSAS. METHODS: The study included 26 patients presenting typical symptoms of sleep apnea. Ten patients were diagnosed to suffer from NARES (mean age 56.8 +/- 12.5, body mass index [BMI] 29.3 kg/m(2) +/- 2.8; 9 men:1 woman) and were compared with 16 age- and BMI-matched individuals (mean age 58.8 +/- 11.6, BMI 29.7 kg/m(2) +/- 3.8, 16 men) without any nasal inflammation, such as allergic rhinitis, sinusitis, nasal polyposis, or vasomotor rhinitis. All patients were tested by polysomnography for an OSAS. RESULTS: Patients suffering from NARES revealed significantly (P <.01) impaired polysomnographic parameters (hypopnea index, apnea-hypopnea index, mean and minimal oxygen saturation) compared with patients without any nasal inflammation. CONCLUSIONS: Our data point to NARES as a risk factor for the induction or augmentation of OSAS. NARES patients suffered from severe OSAS, whereas nondiseased individuals suffered only from moderate OSAS, according to the criteria of the American Academy of Sleep Medicine. Our data support results of others, suggesting chronic nasal inflammation to cause OSAS. Mechanisms for our observations are not fully understood yet. Nasal obstruction or neuronal reflexes might be involved. PMID- 15124167 TI - Inverted papilloma: report of 89 cases. AB - PURPOSE: In this study, the surgical management of an inverted papilloma of the nose and paranasal sinuses performed at our institution is reviewed. METHODS: Eighty-nine patients diagnosed with an inverted papilloma and treated at the Ear, Nose, and Throat Department of Bologna University from January 1980 to January 2001 were retrospectively reviewed. Only 86 out of the 89 patients were retrospectively analyzed because 3 patients were treated for malignant tumors because of the association between inverted papilloma and invasive squamous cell carcinoma. Fifty patients were treated using traditional techniques such as lateral rhinotomy and midface degloving with medial maxillectomy (11 cases), a Caldwell-Luc procedure (21 cases), and transnasal ethmoidectomy or sphenoethmoidectomy (18 cases), whereas 36 patients were treated using an endonasal endoscopic approach for their primary tumor. RESULTS: The traditionally treated patients had a mean follow-up of 8 years (6-19 years), whereas the endoscopically treated patients had a mean follow-up of 54 months (24-97 months). The overall recurrence rate was 15%. There were 12 recurrences (24%) in the traditionally treated patients: 1 out of 11 patients (9%) had a recurrence after a medial maxillectomy by means of a lateral rhinotomy/midface degloving, whereas 11 patients out of 39 (28%) had a recurrence after more conservative procedures such as the transantral approach and transnasal ethmoidectomy or sphenoethmoidectomy. On the other hand, 1 recurrence was observed in the endoscopically treated patients (3%). CONCLUSIONS: Traditional techniques such as Caldwell-Luc and conservative transnasal ethmoidectomy or sphenoethmoidectomy are associated with a high rate of recurrence in the treatment of an inverted papilloma. On the other hand, the endoscopic endonasal approach is an effective treatment for an inverted papilloma in selected cases and, when performed by a skilled surgeon, can achieve a radical resection of the tumor because of excellent magnification and visualization, with results comparable to those of traditional techniques such as lateral rhinotomy with medial maxillectomy. PMID- 15124168 TI - Localized amyloidosis of the tongue: a review. AB - Amyloidosis is a disease entity defined by the presence of extracellular deposits of proteinacious material. These deposits have a characteristic apple-green birefringence with polarized light after staining with Congo red. Amyloid involvement of the tongue is almost universally secondary to systemic disease. The mean survival of patients with a systemic form of amyloidosis is between 5 to 15 months, whereas those with the localized form have an excellent prognosis. This article presents a case of the more unusual localized form. A thorough evaluation, including abdominal fat or rectal biopsy, is essential in every patient to identify any systemic involvement. The absence of systemic amyloidosis offers a much more favorable prognosis and may be treated with simple surgical excision. PMID- 15124169 TI - Paraganglioma presenting as chronic otitis media with cholesteatoma: pitfalls and strategies. AB - The coexistence of paraganglioma with cholesteatoma is a very rare clinical disorder. The clinical presentations are nonspecific. There may be radiological characteristics of either cholesteatoma or paraganglioma in the middle ear area, but the diagnosis of the coexistence of the 2 is usually made only postoperatively. Here is such a case that is made more interesting not only because it initially presented with conductive hearing loss but also because the clinical picture mimicked chronic otitis media. The patient underwent postauricular tympanomastoidectomy with extended facial recess approach to remove the tumor. No evidence of recurrence and complications were noted. PMID- 15124170 TI - Unusual presentation of blunt laryngeal injury with cricotracheal disruption by attempted hanging: a case report. AB - The management of a patient with traumatic disruption of the cricotracheal junction in an attempted suicide by hanging is described. Such injury is uncommon, and many patients die at the scene; detailed radiologic imaging is rare because of the urgency of airway management. The delayed complete disruption of the major airway in this patient allowed adequate imaging and corrective management. Associated soft-tissue injuries of the great vessels of the neck are also described. PMID- 15124171 TI - Cavernous hemangioma of the internal auditory canal: a case report and review of the literature. AB - Cavernous hemangiomas are rare lesions that may be difficult to distinguish from acoustic neuromas, schwannomas, or other lesions of the cerebellopontine angle on imaging studies. The clinical history, workup including radiologic evaluation, histologic assessment, and a brief review of the literature are presented. PMID- 15124172 TI - Pigmented villonodular synovitis of the temporomandibular joint: an extensive case with skull-base involvement. AB - Pigmented villonodular synovitis (PVNS) is a benign lesion of unclear etiology involving the synovial membranes of joints, bursae, and tendon sheaths. Its occurrence in temporomandibular joint (TMJ) is distinctly rare. Despite its benign nature, PVNS is locally destructive to the surrounding structures. Misdiagnosis and delay in diagnosis contribute to the frequency of tumors presenting at an advanced stage at the time of treatment. Thus, PVNS often requires extensive surgery. A case of PVNS of the left TMJ, involving the left infratemporal fossa and subtemporal cranial base, is presented herein. Preoperative computerized tomography, magnetic resonance imaging, and fine-needle aspiration biopsy were used to establish a presumptive diagnosis and to determine the extent of the lesion. Complete excision of the lesion through an infratemporal fossa approach has provided 7 years of a disease-free period with adequate preservation of function. PMID- 15124173 TI - Facial palsy and Wegener's granulomatosis. AB - Facial palsy can occur as a result of various pathological processes, which are not always amenable to early diagnosis. This article is a case presentation of a patient with facial palsy, after an acute otitis media manifestation, as a first symptom of Wegener's granulomatosis. The clues leading to diagnosis consist of the practitioner's clinical suspicion of the disease, the use of the appropriate serological measurements (c-antineutrophil cytoplasmic autoantibody), and the histological confirmation. The early initiation of treatment leads to high rates of remission of an otherwise lethal disease. PMID- 15124174 TI - Allergic fungal rhinosinusitis: a report of 8 cases. AB - Eight patients presented with clinical manifestations such as polyps and mucin were reported to have allergic fungal rhinosinusitis (AFS). Histopathologic sections from tissue samples containing mucin from the paranasal sinuses obtained by endoscopic operation showed scattered hyphal elements within the allergic mucin but no tissue invasion. Associated fungi were 4 cases of Aspergillus flavus; a case each of Aspergillus niger and Bipolaris hawaiiensis, mixed colonization with B. hawaiiensis, and Curvularia lunata; and 1 case of Bipolaris species. Elevated immunoglobulin E level was reported in some patients ranging from (706 to 1969 IU/mL). All patients underwent endoscopic surgery; polypectomy and clearance of all affected sinuses were performed. Medical treatment involved the use of local and systemic corticosteroids. The patients have done well, with no evidence of recurrent disease. PMID- 15124175 TI - Collision tumor of apocrine carcinoma and squamous carcinoma occurring in the ear. AB - An unusual case of a cutaneous collision tumor comprised of a squamous cell carcinoma and an apocrine carcinoma occurring in the auricle of an 82-year-old man is presented. Microscopic study confirmed 2 distinct tumors: a squamous cell carcinoma arising in the epidermis and an adjacent apocrine carcinoma. We discuss the diagnostic criteria and review pertinent recent literature. To our knowledge, this is the first English language report of such a collision tumor. PMID- 15124176 TI - A review of cardiovascular complications accompanying AIDS. AB - Manifestations of cardiovascular system involvement are not uncommon complications of HIV infection, especially in AIDS patients. However, the frequency of these manifestations is influenced by different variables including: survival prolongation in HIV-infected patients, because of advances in antiretroviral treatment; improvement of immunodepression and reduction in the occurrence of opportunistic infections; adverse effects of some drugs. At present, on the whole cardiovascular complications that are HIV correlated in the western world, including Italy, occur less frequently than in the past. However complications associated with alterations in lipometabolism prevail because they can be promoted by some protease inhibitors in predisposed subjects. The most frequently reported questions and a careful analysis of recent data in the medical literature regarding the most common HIV-correlated cardiovascular complications are discussed in this review. PMID- 15124177 TI - Toxicological effects of iron on intestinal cells. AB - The aim of the present study was to investigate whether iron, which is involved in the formation of free radicals in vitro, can initiate cellular injury in human intestinal cells. The effects of various concentrations of iron were studied in preconfluent, colonic-cancerogenous cells, and also in postconfluent, differentiating cells. Cellular damage was assessed using cell proliferation (serial cell counting), tetrazolium dye (MTT) uptake, lactate dehydrogenase (LDH) release and apoptosis studies based on caspase-3 activities. Also the activities of the major antioxidative enzymes, superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) were measured after the cells had been exposed to iron. Our results indicated that preconfluent cells were more susceptible to iron toxicity, as assessed by a significant reduction in cell proliferation and MTT uptake in a concentration-dependent manner compared to the control. However, no evidence for MTT uptake was observed in postconfluent cells. Caspase-3 activity, an indicator of cell apoptosis, considerably increased in preconfluent cells at high iron levels compared to the control (p < 0.05), whereas postconfluent cells were not significantly affected. LDH release was similar for both groups and was significantly higher than the control at 900 microM iron and above. SOD activities were not affected by iron in either group, whereas GPx was considerably higher in iron-treated cells in both groups compared with the control (because of relatively high standard deviations this effect was not significant). In conclusion we suggest that iron exerts its toxic effects intracellularly especially in preconfluent Caco-2 cells, whereas only high iron doses were able to alter the viability of differentiating, enterocyte-like cells. PMID- 15124178 TI - The effect of desferrioxamine on peroxynitrite-induced oxidative damage in erythrocytes. AB - The aim of this study was to investigate the effect of desferrioxamine on peroxynitrite-mediated damage in erythrocytes by measuring the 3-nitrotyrosine level and glutathione peroxidase and Na(+)-K(+) ATPase activities in vitro. 3 Nitrotyrosine levels were determined by HPLC; glutathione peroxidase and Na(+) K(+) ATPase activities were measured by spectrophotometry. Peroxynitrite increased the 3-nitrotyrosine level but decreased both enzyme activities. In the presence of desferrioxamine, glutathione peroxidase activity was increased with a decrease in the 3-nitrotyrosine level. Desferrioxamine was found to possess an important antioxidant activity as assessed in an in vitro system, reducing protein nitration, restoring enzyme activities and maintaining erythrocyte membrane integrity. PMID- 15124179 TI - Alterations of oxidative-antioxidative status in human cutaneous leishmaniasis. AB - Enhanced lipid peroxidation and decreased antioxidant defences have been defined in several diseases. In the present study, we aimed to evaluate the oxidative antioxidative status of patients with cutaneous leishmaniasis (CL). Concentrations of erythrocyte lipid peroxidation (LPO), as an indicator for the oxidative status, reduced glutathione (GSH), glutathione peroxidase (GSH-Px) and serum vitamin C levels, as indicators for the antioxidative status, were measured. Seventy patients aged between 15 and 50 years (38 patients had active CL and 32 patients had healed CL) and 40 healthy controls aged between 19 and 50 years were included in the study. LPO and GSH of the patients with active CL were significantly higher (p < 0.001), whereas erythrocyte GSH-Px and serum vitamin C levels were lower (p < 0.001, p < 0.01 respectively) than those of healthy controls. There was a significant inverse correlation between LPO and serum vitamin C level (r=-0.32, p < 0.05) in active CL. No significant correlation of LPO, GSH, GSH-Px and serum vitamin C levels in control groups or in the group with healed CL was detected. In the light of our findings it is possible to conclude that patients having CL are affected by oxidative stress, which most likely induces the endogenous antioxidant system. An imbalance between the oxidant and antioxidant systems occurs and the suppressed antioxidants and increased lipid peroxidation may contribute to the progression of the disease. PMID- 15124180 TI - In vivo evidence suggesting a role for purine-catabolizing enzymes in the pathogenesis of cisplatin-induced nephrotoxicity in rats and effect of erdosteine against this toxicity. AB - The aim of this experimental study was to investigate the possible role of adenosine deaminase (AD) and xanthine oxidase (XO) in the pathogenesis of cisplatin-induced nephrotoxicity and the effect of erdosteine in decreasing the toxicity. The intraperitoneal injection of cisplatin (7 mg kg(-1) body weight) induced a significant increase in plasma creatinine level and blood urea nitrogen (BUN), and plasma and damaged renal tissue activities of AD and XO in rats. Co treatment with erdosteine (10 mg kg(-1)day(-1)) attenuated the increase in the plasma creatinine and BUN levels, and significantly prevented the increase in tissue and plasma AD and XO activities (P<0.05). The results of this study revealed that XO and AD may play an important role in the pathogenesis of cisplatin-induced nephrotoxicity. The potent free radical scavenger erdosteine may have protective potential in this process and it will become a promising drug in the prevention of this undesired side-effect of cisplatin, but further studies are needed to illuminate the exact protection mechanism of erdosteine against cisplatin-induced nephrotoxicity. PMID- 15124181 TI - Paraoxonase 55 and 192 polymorphism and its relationship to serum paraoxonase activity and serum lipids in Turkish patients with non-insulin dependent diabetes mellitus. AB - We investigated the effect of PON 55 and PON 192 polymorphisms on serum PON1 activity and lipid profiles in 213 non-insulin dependent diabetes mellitus (NIDDM) individuals and 116 non-diabetic controls among Turkish subjects. The distribution of PON 55/192 gene polymorphism was determined by polymerase chain reaction-based restriction fragment length polymorphism. Serum lipid levels were measured enzymically. PON activity was measured by spectrophotometric assay of p nitrophenol production following addition of paraoxon. We found that PON 55 and 192 genotype distribution was similar in patients and controls and paraoxonase activity was generally lower in diabetics than in control subjects. We showed that PON 55 and 192 genotypes have a major effect on serum PON activity. PON 192 BB homozygotes had significantly higher PON activity than AA and AB genotypes among the control and NIDDM populations (p<0.001). PON 55 MM homozygotes had significantly lower PON activity than did LL and LM genotypes in control and NIDDM populations (p<0.05). The PON1 55 and 192 polymorphisms did not consistently influence the serum lipid profiles in either population. In conclusion, our results suggest that the paraoxonase activities are affected by PON1 genetic variability in Turkish NIDDM patients and controls. PMID- 15124182 TI - Amelioration of methotrexate-induced enteritis by melatonin in rats. AB - The anti-tumour drug methotrexate (MTX) induces intestinal mucosa injury resulting in malabsorption and diarrhoea. The purpose of this study was to investigate whether exogenous melatonin could protect the gut from MTX-induced damage in rats. A single dose of MTX (20 mg kg(-1), i.p.) was followed by i.p. saline or melatonin injections (10 mg kg(-1), MTX + Mel) for the next 5 days. On the fifth day, intestinal transit was assessed using charcoal propagation. Rats were decapitated and small intestinal segments were fixed for light (LM) and scanning electron microscope (SEM) examinations. Other intestinal segments were stored to measure glutathione (GSH) and malondialdehyde (MDA) levels, myeloperoxidase (MPO) and ATPase activity. MTX led to loss of more than 10% of the initial body weight (p < 0.01). Conversely, weight loss was markedly less in the melatonin-treated MTX group (p < 0.05). Bowel motility was increased in MTX treated rats, while the transit index in the MTX-Mel group was not different from the control group. MTX caused decreases in GSH levels and ATPase activity, with increases in MDA levels and MPO activity. These changes were reversed in MTX-Mel treated rats (p < 0.05-p < 0.001). LM and SEM in the MTX group revealed desquamation of surface epithelium and glandular degeneration, while the epithelium was slightly damaged in the MTX-Mel group. In conclusion, the present study demonstrates that melatonin is capable of reversing MTX-induced intestinal dysfunctions, indicating that it may be beneficial in ameliorating the symptoms of chemotherapy-induced enteritis. PMID- 15124183 TI - The interaction of haemoglobin, magnesium, organic phosphates and band 3 protein in nucleated and anucleated erythrocytes. AB - The anion influx was measured in order to study the interaction among organic phosphates, magnesium, haemoglobin and the N-terminal of the cytoplasmic domain of band 3 protein in human, chicken and trout erythrocytes. The rate constant for SO(4)(2-) influx in human and trout erythrocytes increased significantly when it was measured with an increased concentration of intracellular Mg(2+). The SO(4)(2 ) influx was also measured in human erythrocyte ghosts in the presence and absence of Mg(2+). The smaller activation provoked by Mg(2+) in ghosts could be caused by the presence of a small quantity of haemoglobin which remained inside. The SO(4)(2-) uptake in chicken erythrocytes in the presence and in absence of Mg(2+) was characterized by very similar rate constants. The results suggest that the small increase in intracellular Mg(2+) in the erythrocytes involves an increase in the formation of Mg(2+)-ATP and Mg(2+)-2,3 BPG complexes reducing the affinity of the organic phosphates for Hb. This new situation may influence the functions of the anion transporter with consequent variations of SO(4)(2-) influx throughout the erythrocyte membrane in human and in trout erythrocytes, whereas in chicken RBCs this function cannot occur and, in fact, no increase in sulphate influx was noticeable. The measurement of Hb/O(2) affinity by the use of alternating fixed and variable concentrations of organic phosphates and Mg(2+), confirms the interactions between these elements and their effect on the mechanism of the affinity. When we measured the sulphate influx in the presence of DIDS we found some differences in the three types of cells. PMID- 15124184 TI - 5-Fluorouracil enhances apoptosis sensitivity of T lymphocytes mediated by CD3 epsilon. AB - Previous studies by our laboratory have reported that the T cell receptor (TCR) TCR/CD3 complex could mediate activation as well as apoptosis of T lymphocytes. Two tyrosine residues in the ITAM (immuno-receptor tyrosine-based activation motifs) of CD3 epsilon were required for apoptosis signalling of Jurkat T lymphocytes. Stable cell lines TJK and T3JK produced from CD8(-) Jurkat T lymphocytes by transfection with wild-type and mutant CD8 epsilon (fusion of the extracellular and transmembrane domains of human CD8 alpha to the intracellular domain of mouse CD3 epsilon), were used with CD8(-) Jurkat T lymphocytes for studying the role of single intact CD3 epsilon. 5-Fluorouracil (5-FU), a chemotherapeutic drug can induce cell death of many tumour cell lines. In the present experiments, we examined the expression of caspase-3, p53 and Bid in the three cell lines induced by 5-FU and/or anti-CD8 antibody. We found high expression of p53 during activation-induced cell death of TJK cells mediated by anti-CD8 antibody and apoptosis of TJK and T3JK induced by 5-FU, implicating p53 involvement in apoptosis of leukemia cells induced by anti-CD8 antibody and 5-FU. We also detected the active form of caspase-3 and Bid in apoptotic leukemia cells after treatment with 5-FU and/or anti-CD8 antibody, indicating that the drug and antibody induced cell death through caspase-3 and the signal pathway may involve the Bcl-2 protein family. Our results showed that combined treatment with 5-FU and anti-CD8 antibody could enhance the rate of apoptosis induced by 5-FU or anti CD8 antibody through increased expression of p53 and by promoting activation of caspase-3 and Bid. This suggests that the combination of 5-FU and anti-CD8 antibody may play an important role in inducing apoptosis of leukemia cells. PMID- 15124185 TI - Effect of endorphin exposure at weaning on the endorphin and serotonin content of white blood cells and mast cells in adult rat. AB - Hormonal imprinting takes place perinatally at the first encounter between the developing receptor and its target hormone, resulting in the accomplishment of normal receptor development. In the presence of an excess of target hormone or the absence of it, or an excess of related molecules which can be bound by the receptor, faulty imprinting develops with life-long consequences. In previous experiments neonatal endorphin exposure caused a decrease in endorphin and serotonin content of peritoneal mast cells of adult animals. In the present experiment 25-day-old (weaned) female rats received 2 microg endorphin, and the endorphin as well as serotonin content of adult mast cells and white blood cells was studied by flow cytometry and confocal microscopy. Peritoneal lymphocytes and blood monocytes contained significantly (p<0.01) less endorphin and peritoneal mast cells less serotonin (p<0.07, i.e. of questionable significance) than the untreated control. The results bring attention to the possibility of durable imprinting of differentiating cells later in life and to the durable (possibly life-long) effect of an endorphin excess (perhaps caused by injury) manifested in the change of endorphin and serotonin content of immune cells. PMID- 15124186 TI - Prolonged effect of an H1-receptor blocker antihistamine on the histamine content of white blood cells and mast cells. AB - Hormonal imprinting usually takes place perinatally at the first encounter between the developing receptor and its target hormone, determining the future binding capacity of the receptor for life. Molecules similar to a hormone can cause faulty imprinting also with life-long consequences. Hormone production of the imprinted cell is also durably influenced. In cytogenic organs imprinting can also be provoked in adulthood. At present the effect of a single terfenadine treatment in adult rats on the histamine content of peritoneal cells (lymphocytes, mast cells and the monocyte-macrophage-granulocyte group), white blood cells (lymphocytes, granulocytes, monocytes) and thymic lymphocytes was studied 3 weeks after treatment to clarify the effect of prolonged treatment with an antihistamine in adulthood. The cells were studied by flow cytometric analysis. Peritoneal mast cells contained significantly more and thymic lymphocytes significantly less histamine than controls. In the other cells the differences were not significant. The results support earlier observations on the effect of antihistamines on mast cell histamine release (inhibition) and call attention to the fact that this effect is durable (hormonal imprinting provoked in adults). PMID- 15124187 TI - Combination of endometrial thickness and time since menopause in predicting endometrial cancer in women with postmenopausal bleeding. AB - PURPOSE: This study was conducted to assess the combination of endometrial thickness, as measured by transvaginal sonography, and time since menopause, in predicting the presence of endometrial cancer in women with postmenopausal bleeding. METHODS: The study group consisted of 95 women with postmenopausal bleeding who underwent sonographic measurement of endometrial thickness followed by endometrial biopsy. No patient had ever received hormone replacement therapy. RESULTS: The mean endometrial thickness was significantly lower in the absence of endometrial carcinoma (6.9 +/- 4.3 mm) than in its presence (13.5 +/- 7.7 mm) (p < 0.005). The incidence of endometrial carcinoma increased with increases in endometrial thickness and the number of years since menopause. No patient had carcinoma when the endometrium was less than 5 mm thick, but 18.5% did when the thickness exceeded 9 mm. The incidence of cancer was 2.6% in women who had undergone menopause less than 5 years earlier but was 21.4% in women who had undergone menopause more than 15 years prior. Multiple logistic regression analysis showed that time since menopause and endometrial thickness were statistically significant predictors of endometrial carcinoma. CONCLUSIONS: Time since menopause and endometrial thickness together define cutoff points for the diagnostic biopsy of tissue samples for endometrial carcinoma; that is, within a particular time interval, sampling should not be performed if the thickness is below a given value. When using cutoff points of 6 mm of endometrial thickness for women experiencing menopause 5-15 years prior and 5 mm in those going through menopause 15 or more years prior, approximately 60% of invasive procedures may be avoided. In addition, models derived by multiple logistic regression can be used to calculate a patient's risk of cancer based on her age and endometrial thickness. PMID- 15124188 TI - Comparison of transperineal and transvaginal sonography in predicting preterm delivery. AB - PURPOSE: A major advantage of transperineal sonography (TPUS) is its ability to evaluate the cervix without causing any distortion. This study was performed to compare transvaginal sonography (TVUS) and TPUS at 24 weeks of gestation in predicting preterm delivery in low-risk pregnancy. METHODS: Three hundred fifty seven pregnant women underwent TVUS and TPUS at 24 weeks of gestation. The relationship between cervical length and preterm delivery was assessed. Accuracy values of TVUS and TPUS at 24 weeks of gestation were compared in predicting preterm delivery. RESULTS: Preterm delivery (before 36 weeks of gestation) occurred in 22 pregnancies (6.2%). Mean cervical lengths measured by TVUS and TPUS were significantly different in preterm and term delivery groups (P < 0.05). Areas under the curves were 0.801 and 0.857 for the transvaginal and transperineal measurements, respectively. The coefficient of correlation between the transvaginal and transperineal cervical length measurements was 0.83. TPUS had a sensitivity of 77% in predicting preterm delivery, with a false-positive rate of 17% and a relative risk of 4.5 at the 32.5-mm cutoff value. CONCLUSIONS: When the cervix is well visualized, TPUS can predict preterm delivery as accurately as TVUS. PMID- 15124189 TI - Placental thickness in the first half of pregnancy. AB - PURPOSE: This study was conducted to establish normal values of placental thickness during the first half of pregnancy. METHODS: Normal pregnant women with singleton pregnancies between 8 and 20 weeks of gestation were recruited into the study. All the newborns were normal at birth. Placental thickness was measured perpendicularly through the thickest part of the placenta on transabdominal scans. The placental thickness data were analyzed for mean, standard deviation, 95% confidence interval, and 2.5(th), 5(th), 50(th), 95(th), and 97.5(th) percentile for each week of gestational age. The best-fit mathematical model was derived by regression analysis. RESULTS: The total number of measurements was 333 and the number of measurements for each week of gestational age ranged from 9 to 37. Regression analysis yielded the following linear equation of the relationship: placental thickness (in mm) = gestational age (in weeks) x 1.4-5.6 (r = 0.82). CONCLUSION: We have established a nomogram for placental thickness. This resource may be a useful aid in the early detection of placental abnormalities, such as hydropic placenta secondary to hemoglobin Bart's disease. PMID- 15124190 TI - Sonographic assessment of clubfoot. AB - PURPOSE: This study was performed to develop a standardized methodology for the sonographic assessment of clubfoot at birth and at the end of both conservative treatment and surgical correction. METHODS: Forty-two congenital clubfeet and 42 normal feet were examined sonographically in the position of spontaneous alignment and during passive manual correction. Scans along 4 planes provided information relevant to the assessment: sagittal posterior, sagittal anterior, coronal lateral, and transverse. RESULTS: Sagittal posterior sonograms demonstrated the progressive gain of dorsiflexion ability during the different steps of treatment for clubfoot. Sagittal anterior sonograms could not demonstrate the normal alignment of the navicular in clubfeet because of the bone's medial displacement. On transverse sonograms, the talar head and the medially displaced navicular may lie on the same plane, depending on the severity of the deformity. Coronal lateral sonograms provided for estimation of the relationships between the calcaneus and cuboid, which were described by the calcaneal-cuboid angle. CONCLUSIONS: Sonography is a promising technique for assessment and monitoring of clubfoot during treatment. The method described here yields accurate and reproducible information about the anatomy of the nonossified clubfoot, helping the orthopedic team decide on appropriate treatment steps. PMID- 15124191 TI - B-flow imaging of internal carotid artery stenosis: Comparison with power Doppler imaging and digital subtraction angiography. AB - PURPOSE: Digital subtraction angiography (DSA) is the gold standard in the diagnosis of carotid artery stenosis, but it has a relatively high complication rate. We evaluated the efficacy of B-flow imaging (BFI) in examining internal carotid artery stenosis (ICAS) compared with power Doppler imaging (PDI) and DSA. METHODS: We performed BFI, PDI, and DSA on 56 consecutive patients with suspected ICAS. The degree of stenosis was calculated for each technique, and results of BFI and PDI were then correlated with those of DSA. RESULTS: Measurements of the percentage of stenosis made using both sonographic techniques were significantly correlated with those of DSA (p < 0.0001). However, the coefficient of correlation between DSA and BFI (r = 0.94) was higher than that between DSA and PDI (r = 0.87). The mean difference between ICAS measurements with BFI and DSA was -1.3% (95% confidence interval [CI], -2.5 to 0). The mean difference between ICAS measurements with PDI and DSA was -6.5% (95% CI, -8.2 to -4.7). ICAS was graded significantly lower on PDI than on DSA, whereas BFI findings were similar to those of DSA. CONCLUSION: BFI shows high correlation with DSA and provides a more accurate planimetric evaluation of ICAS than PDI does. PMID- 15124192 TI - Sonographic demonstration of duodenobiliary reflux with soda enhancement. AB - Cholangitis rarely occurs after sphincteroplasty if there is no biliary obstruction. We report the case of a patient who developed recurrent cholangitis despite having a patent biliary tract after sphincteroplasty. Duodenobiliary reflux was demonstrated on sonography after enhancement of the reflux flow with a novel oral contrast agent, a carbonated soda beverage. Sonography with contrast enhancement provided by soda solution may prove satisfactory to detect duodenobiliary reflux after sphincteroplasty safely, effectively, and economically. PMID- 15124193 TI - Infertility due to intrauterine residual fetal bone fragments. AB - A case of intrauterine retention of fetal bone diagnosed 8 years after termination of a pregnancy is presented. The patient had a history of hypermenorrhea infertility, and persistent vaginal discharge beginning after the abortion. Transvaginal sonography demonstrated an intrauterine foreign body. Curettage was performed, and fetal bone fragments were found within the removed materials. PMID- 15124194 TI - Prenatal sonographic diagnosis of acrania associated with amniotic bands. AB - We describe the prenatal sonographic findings in 2 cases of amniotic band sequence-related acrania. In the first case, sonography demonstrated acrania, asymmetric facial clefts, and digital fusion on the hands and feet. In the second case, sonography demonstrated acrania and amniotic bands adhering to the brain, face, and body. This report highlights the importance of a careful search for amniotic bands and a detailed survey of the fetus' face, digits, and body in the sonographic evaluation of fetal acrania. PMID- 15124195 TI - Transperineal sonographic findings in a woman with urethral mucosa prolapse. AB - We present a case of urethral mucosa prolapse in a postmenopausal woman. Transperineal gray-scale sonograms revealed a clover-shaped, hypoechoic mass protruding from the urethral meatus. Doppler studies revealed blood flow within the protruding mass. Detection of feeding arteries and draining veins contributed to exclude the possibility of necrosis and gangrene, thus allowing conservative medical treatment to be tried. The mucosal prolapse regressed after 1 week's treatment with topical estrogen and antibiotics and sitz baths. Follow-up sonograms showed a normal urethral configuration and diminished vascular flow when compared with the initial study. PMID- 15124196 TI - Late-onset ureteric urinoma with intermittent jet flow as a complication of ureterolithotomy. AB - We present the case of a late-onset urinoma in a 47-year-old man with a history of right ureterolithotomy performed 6 months previously, following 2 unsuccessful attempts at extracorporeal shock wave lithotripsy. The ipsilateral kidney was nonfunctional and hydronephrotic. The urinoma did not show enhancement on contrast-enhanced CT and intravenous pyelography. Sonography revealed turbulence in the urinoma and jet flow from the ureter. Lab studies of fluid obtained by percutaneous puncture and aspiration yielded misleading findings for urine and revealed the presence of hemosiderin-laden macrophages. The urinoma was surgically resected and ureteroneocystostomy was performed. PMID- 15124197 TI - Summary workshop report: biopharmaceutics classification system--implementation challenges and extension opportunities. PMID- 15124198 TI - Assessing the antifungal activity and toxicity profile of amphotericin B lipid complex (ABLC; Abelcet) in combination with caspofungin in experimental systemic aspergillosis. AB - The purpose of this study was to assess the antifungal activity and renal and hepatic toxicity of amphotericin B lipid complex (ABLC; Abelcet) following co administration of Caspofungin to rats infected with Aspergillus fumigatus. Aspergillus fumigatus inoculum (1.3-2.3 x 10(7) colony forming units [CFU]) was injected via the jugular vein; 48 h later male albino Sprague-Dawley rats (350 400 g) were administered either a single intravenous (i.v.) dose of Fungizone(R) (1 mg AmpB/kg), ABLC (1 or 5 mg AmpB/kg), or an equivalent volume of normal saline (NS) (vehicle control) once daily for 4 days. Rats were further randomized into groups to receive 3 mg/kg Caspofungin or physiologic saline i.v. once daily for 4 days. To assess antifungal activity, brain, lung, heart, liver, spleen, and kidney sections were homogenized with NS (2 mL; 1 g of each tissue/mL) and a 0.1 mL aliquot was spread plated onto a Sabouraud dextrose agar plate. The plates were incubated for 48 h at 37 degrees C, at which time the numbers of CFU were determined and corrected for tissue weight. To assess renal and hepatic toxicity, serum creatinine and aspartate aminotransferase levels were determined. Fungizone and ABLC at a dosing regimen of 1 mg/kg i.v. once daily for four consecutive days and Caspofungin at a dosing regimen of 3 mg/kg i.v. once daily for four consecutive days had similar effectiveness in decreasing the total number of Aspergillus fumigatus CFUs found in all organs analyzed compared to non-treated controls. A combination of ABLC (1 mg/kg i.v. x 4 days) and Caspofungin (3 mg/kg i.v. x 4 days) significantly decreased the total number of Aspergillus fumigatus CFUs found in all organs analyzed compared to Caspofungin alone and non-treated controls. ABLC at a dosing regiment of 5 mg/kg i.v. once daily for four consecutive days was more effective in decreasing the total number of Aspergillus fumigatus CFUs found in all organs analyzed compared to Fungizone or ABLC alone at 1 mg/kg and Caspofungin alone at 3 mg/kg. However, a combination of ABLC (5 mg/kg i.v. x 4 days) and Caspofungin (3 mg/kg i.v. x 4 days) was not more effective than ABLC at 5 mg/kg or the combination of ABLC at 1 mg/kg and Caspofungin 3 mg/kg in reducing the total number of Aspergillus fumigatus CFUs compared to controls. Except for non-treated infected control rats, none of the treatment groups tested displayed a greater than 50% increase in serum creatinine concentrations from baseline. In addition, only ABLC at a dosing regimen of 1 mg/kg i.v. once daily for four consecutive days displayed a greater than 50% increase in AST concentration from baseline. Taken together, these findings suggest that ABLC at 5 mg/kg once daily x 4 days appears to be the best therapeutic choice in this animal model. PMID- 15124199 TI - Challenges in the development of high protein concentration formulations. AB - Development of formulations for protein drugs requiring high dosing (in the order of mg/kg) may become challenging for solubility limited proteins and for the subcutaneous (SC) route with <1.5 mL allowable administration volume that requires >100 mg/mL protein concentrations. Development of high protein concentration formulations also results in several manufacturing, stability, analytical, and delivery challenges. The high concentrations achieved by small scale approaches used in preformulation studies would have to be confirmed with manufacturing scale processes and with representative materials because of the lability of protein conformation and the propensity to interact with surfaces and solutes which render protein solubilities that are dependent on the process of concentrating. The concentration dependent degradation route of aggregation is the greatest challenge to developing protein formulations at these higher concentrations. In addition to the potential for nonnative protein aggregation and particulate formation, reversible self-association may occur, which contributes to properties such as viscosity that complicates delivery by injection. Higher viscosity also complicates manufacturing of high protein concentrations by filtration approaches. Chromatographic and electrophoretic assays may not accurately determine the non-covalent higher molecular weight forms because of the dilutions that are usually encountered with these techniques. Hence, techniques must be used that allow for direct measurement in the formulation without substantial dilution of the protein. These challenges are summarized in this review. PMID- 15124200 TI - A novel method for the preparation of liposomes: freeze drying of monophase solutions. AB - A novel method is described for the preparation of sterile and pyrogen-free submicron liposomes of narrow size distribution. The method is based on the formation of a homogeneous dispersion of lipids in water-soluble carrier materials. To obtain the lipid-containing solid dispersion, liposome-forming lipids and water-soluble carrier materials are dissolved in tert-butyl alcohol/water cosolvent systems to form an isotropic monophase solution, and then the resulting solution is lyophilized after sterilization by filtration through 0.2 microm pores. On addition of water, the lyophilized product spontaneously forms homogeneous liposome preparation. After investigation of the various parameters associated with this method it is found that the lipid/carrier ratio is the key factor affecting the size and the polydispersity of liposome preparation. Based on the data from DSC, X-ray diffraction, and size measurements, a possible liposome formation mechanism is proposed. In addition, the application of this new method to the passive loading and active loading of drugs into liposomes is discussed in detail. PMID- 15124201 TI - Mechanistic study of chemical skin permeation enhancers with different polar and lipophilic functional groups. AB - In a previous study, the enhancement effects on the transport of a steroidal permeant along the hairless mouse skin (HMS) stratum corneum (SC) lipoidal pathway were investigated for two homologous series of chemical enhancers: the 1 alkyl-2-pyrrolidones and the 1-alkyl-2-azacycloheptanones. The objective of the present study was to extend this investigation to a broader range of enhancers in order that generalizations with regard to the mechanistic aspects of enhancer function might be established. Specific questions to be addressed included: (a) what is the nature of the microenvironment of the enhancer site of action? (b) what is the extent of the equilibrium uptake of the enhancer from its E = 10 aqueous enhancer solution (the aqueous concentration for which the enhancer induces a tenfold transport enhancement) into the HMS SC intercellular lipid "phase"? and (c) are the microenvironment of the enhancer site of action and that for the equilibrium enhancer uptake at E = 10 relatively independent of the molecular characteristics of the enhancers (as suggested by the earlier study)? Enhancers selected for this study included: a wide range of polar head group size and polarity; n-alkyl group chain lengths from C(4) to C(12); and enhancers in which a double bond is substituted for a single bond in the hydrocarbon chain (3 alkenols) from C(5) to C(9). In addition to the main study, an ancillary set of experiments were to be conducted on the partitioning of a surrogate permeant (estradiol) into the intercellular lipid "phase" under E = 10 isoenhancement conditions to assess the extent to which the permeant partition coefficient may contribute to the permeation enhancement. The following were the principal findings of this research. First, there was very good correlation between the E = 10 isoenhancement aqueous enhancer concentrations and K(octanol/water) for all the studied enhancers. Second, the partitioning of the enhancer from the E = 10 aqueous enhancer solution into the HMS SC intercellular lipid "phase" was found to be relatively independent of the molecular characteristics for all studied enhancers, and the partition coefficients also correlated well with K(octanol/water). These results may have the following meanings: both the microenvironment of the enhancer site of action and the SC intercellular lipid "phase" involved in the enhancer partitioning experiments are well mimicked by liquid n-octanol, and the "intrinsic" potencies (as assessed by the equilibrium enhancer concentration in the microenvironment at the site of action) of the enhancers are relatively independent of the molecular characteristics of the studied enhancers. Finally, the estradiol partitioning experiments suggest the permeant partitioning into the HMS SC intercellular lipid "phase" is enhanced around five- to seven-fold when permeation is enhanced ten-fold for most of the studied enhancers; therefore, the enhancement of the permeant partition coefficient rather than the permeant diffusion coefficient seems to be more important in permeation enhancement of the SC barrier lipoidal pathway. PMID- 15124202 TI - Oral bioavailability and multiple dose tolerability of an antisense oligonucleotide tablet formulated with sodium caprate. AB - In vivo study was performed to determine the tolerability and pharmacokinetics of ISIS 104838, a phosphorothioate antisense oligonucleotide targetting human tumour necrosis factor alpha (TNF-alpha) mRNA, following multi-dose administration via intravenous and oral routes. Oral tablet formulations of ISIS 104838 were pre formulated with the permeation enhancer, sodium caprate, in an enteric-coated solid dosage form. The average plasma bioavailability of ISIS 104838 was 1.4% relative to IV. The tissue distribution profile was similar following both routes of administration, with highest concentrations observed in the kidney followed by the liver, lymph nodes and spleen. Plasma bioavailability underestimated the tissue accumulation of ISIS 104838 observed 1 day after the last dose. Mean systemic tissue bioavailability ranged from 2.0 to 4.3%, relative to IV tissues, and was dependent on tissue type. No marked differences were noted in the pharmacokinetic parameters following multi-dosing either via intravenous or oral routes. All formulations administered were well tolerated. This paper reports the first evaluation of solid oral dosage forms comprising sodium caprate and an antisense oligonucleotide. Furthermore, this study demonstrates the oral delivery of ISIS 104838 from solid oral dose formulations, with the achievement of comparable tissue concentrations of the oligonucleotide to that of the intravenous treatment. PMID- 15124203 TI - Combined application of parallel artificial membrane permeability assay and Caco 2 permeability assays in drug discovery. AB - Data from permeability profiling using the parallel artificial membrane permeability assay (PAMPA) and cell monolayer (Caco-2 and MDR1-MDCKII) methods were compared for two published compound sets and one in-house set. A majority of compounds in each set correlated (R(2) = 0.76-0.92), indicating the predominance of passive diffusion in the permeation of these compounds. Compounds that did not correlate grouped into two subsets. One subset had higher PAMPA permeability than cell monolayer permeability and consisted of compounds that are subject to secretory mechanisms: efflux or reduced passive diffusion of bases under Caco-2 when run under a pH gradient. The other subset had higher cell monolayer permeability than PAMPA permeability and consisted of compounds that are subject to absorptive mechanisms: paracellular, active transport, or increased passive diffusion of acids under Caco-2 when run under a pH gradient. Given the characteristics of the two methods, these studies suggest how PAMPA and Caco-2 can be synergistically applied for efficient and rapid investigation of permeation mechanisms in drug discovery. During early discovery, all compounds can be rapidly screened using PAMPA at low pH and neutral pH to assess passive diffusion permeability to indicate potential for gastrointestinal and cell assay permeation. During intermediate discovery, selected compounds can be additionally assayed by apical-to-basolateral Caco-2, which, in combination with PAMPA data, indicates susceptibility to additional permeation mechanisms (secretory and absorptive). During mid-to-late discovery, selected candidates can be examined in detail via multiple directional Caco-2 experiments and with transporter inhibitors for complete characterization of permeation mechanisms. PMID- 15124204 TI - Drying of poloxamer hydrogel films. AB - The drying of hydrogel films formed by Poloxamer 407 poly(ethylene oxide) poly(propylene oxide)-poly(ethylene oxide) amphiphilic block copolymer was investigated at various air relative humidity (RH) conditions ranging from 11 to 97%. Initially, the amount of water lost increased linearly with the drying time. After this linear region (stage I), a nonlinear behavior was observed (stage II). The drying rate increased with decreasing RH, thus greatly shortening the drying time. A decrease of the film thickness also shortened the drying time; however, the drying mechanism did not change. Three models for one-dimensional water diffusion were used to fit the experimental results at different RH conditions and film thicknesses. Model 1 assumes semi-infinite medium and constant diffusion coefficient, and fits very well the data in stage I of the drying process. The fitted water diffusion coefficient (D) is 5 x 10(-10) m(2)/s, whereas the effects of the RH are captured by a proportionality constant (alpha) that appears in the boundary condition. Model 2 considers a finite (constant) film thickness and captures the experimental observations over the whole drying period for the same D and alpha as in Model 1. The analytical solutions available for Models 1 and 2, used together with the experimentally derived model parameters D and alpha, allow for easy estimation of drying time and water loss from Poloxamer hydrogel films of various compositions and thicknesses and at different relative humidities. Numerical solutions for water diffusion under conditions of decreasing film thickness and diffusion coefficient being a function of concentration are also presented (Model 3). It becomes apparent from the fit of the data to the different models that the drying rate is more sensitive to the boundary condition at the film-air interface (represented by alpha) than to the diffusion in the film. It is notable that the alpha values obtained from the fits of the Poloxamer hydrogel drying rate are comparable to those obtained from drying of water films under the same experimental conditions. PMID- 15124205 TI - Solubilization behavior of poorly soluble drugs with combined use of Gelucire 44/14 and cosolvent. AB - Gelucire 44/14 is a surface-active excipient that can solubilize poorly soluble drugs. We investigated its solubilization behavior when coexisting with dimethylacetoamide (DMA) or dimethylsulfoxide (DMSO), both of which are also expected to enhance drug solubility. Gelucire was confirmed to form micelles by surface tension and fluorescence measurements both in water and water/cosolvent mixtures. Light-scattering measurements revealed that DMA and DMSO affect the micellar morphology in a different manner. DMA helped form large structures by being entrapped in the hydrophobic region of the micelles and/or inducing the aggregation. DMSO was likely to be anchored to the interfacial layer and did not induce micelle growth. Two model drugs, phenytoin and indomethacin, were employed to observe the solubilization behavior of poorly soluble drugs in Gelucire/cosolvent mixtures. The solubility of these drugs in the mixtures could be explained very well by using the new solubility model introduced in this article. Addition of cosolvents to the Gelucire solution did not enhance the solubility very much, and thus the combined use of cosolvents with Gelucire offered only little advantage from the viewpoint of solubility. PMID- 15124206 TI - Computational prediction of the plasma protein-binding percent of diverse pharmaceutical compounds. AB - A nonlinear regression analysis has been applied to develop a new method to predict the plasma protein-binding percent of structurally diverse pharmaceutical compounds. The analysis included over 300 launched drugs with experimental human plasma protein binding percent data. These drugs were classified according to protonation state and pharmacophore features. The correlation formula for each class is a simple sigmoidal function of variable LogP or LogD. A correlation formula of variable LogD at pH 7.4 with a good correlation coefficient (R-squared = 0.803) was obtained for neutral and basic drugs, with the exception of zwitterions. A correlation formula using LogP as variable for acidic drugs with one of the specific pharmacophore features gave a good correlation coefficient (R squared = 0.786). The method was verified using the protein binding data of 20 compounds that had not been included in the data set to configure the formulas. The correlation coefficient (R-squared) between the experimental and predicted protein binding percent was 0.830. In conclusion, the method developed and described in this report can provide precise and useful prediction of plasma protein binding percent for new drug candidates. PMID- 15124207 TI - Comparative bioavailability study in dogs of a self-emulsifying formulation of progesterone presented in a pellet and liquid form compared with an aqueous suspension of progesterone. AB - A pellet formulation of progesterone in a self-emulsifying system (SES) was prepared by the process of extrusion/spheronization to provide a good in vitro drug release (100% within 30 min, T(50%) at 13 min). A three-way randomized crossover study was performed in six fasted male beagle dogs with these pellets and the same SES liquid formulation, both contained in a hard shell capsule, and an aqueous suspension. The same dose of progesterone (16 mg) in pellets and in the SES liquid formulation resulted in similar AUC, C(max) and T(max) values, estimated from progesterone plasma levels by (125)I radioimmunoassay. Although the maximum absorption was slightly retarded (0.5 to 1 h) by SES (pellets and liquid), AUC and C(max) were approximately seven and nine times greater then those obtained when an aqueous suspension formulation of the same dose of progesterone was administered to the same dogs. These results showed that it was possible to improve the bioavailability of the poorly soluble, poorly permeable progesterone when administered in SES. Moreover, presenting the progesterone in the form of a pellet did not prevent the release of the drug in vivo. These data demonstrate the utility of extrusion/spheronization in delivering a nonaqueous system in a novel solid dosage form. PMID- 15124208 TI - Estimation of the fragility index of indomethacin by DSC using the heating and cooling rate dependency of the glass transition. AB - In this study we have investigated the features of the glass transition relaxation of indomethacin using Differential Scanning Calorimetry (DSC). The purpose of this work is to provide an estimation of the activation energy at the glass transition temperature, as well as of the fragility index, of amorphous indomethacin from DSC data. To do so, the glass transition temperature region of amorphous indomethacin was characterized in both cooling and heating regimes. The activation energy for structural relaxation (directly related to glass fragility) was estimated from the heating and cooling rate dependence of the location of the DSC profile of the glass transition. The obtained results were similar in the heating and in the cooling modes. The results on the fragility index of indomethacin obtained in the present study, m = 60 in the cooling mode and m = 56 in the heating mode, are compared with other values previously published in the literature. PMID- 15124209 TI - Human skin permeation and partition: general linear free-energy relationship analyses. AB - Literature values of the permeability coefficient for permeation of human skin from water have been adjusted for ionization in water and adjusted for temperature. The obtained values of log K(p) for 119 solutes at 37 degrees C have been correlated with Abraham descriptors to yield an equation with R(2) = 0.832 and SD = 0.46 log units. Three separate test sets of 60 compounds had log K(p) predicted with an SD of 0.48 log units. The main factors that influence log K(p) are solute hydrogen bond basicity that lowers the permeability coefficient and solute volume that increases the permeability coefficient. Human skin-water partition coefficients, as log K(sc), have been collected for 45 compounds and yield an equation with R(2) = 0.926 and SD = 0.22 log units. We have compared the log K(p) equation to equations for various other processes, but have found no process that appears to be similar to that for skin permeation. The nearest process to skin-water partition is the isobutanol-water partition system. An equation for lateral diffusion in the stratum corneum is shown to be reasonably close to various diffusion-related processes. PMID- 15124210 TI - Effect of tumor necrosis factor-alpha and interferon-gamma on intestinal P glycoprotein expression, activity, and localization in Caco-2 cells. AB - The P-glycoprotein (Pgp), a drug efflux pump, is expressed in intestinal epithelial cells, where it constitutes a barrier against xenobiotics. In inflammatory bowel disease, a dysregulation in the production of tumor necrosis factor (TNF)alpha and interferon (IFN)gamma, and an alteration of Pgp expression and activity have been reported. The aim of this study was to investigate the effects of TNF alpha and IFN gamma on intestinal Pgp expression, activity, and localization in Caco-2 cells grown on filters. TNF alpha induced both a strong time-dependent diminution (-56%) of MDR1 mRNA (semiquantitative reverse transcription polymerase chain reaction) and a significant decrease of unidirectional transport of rhodamine 123 after 48 h of exposure at 10 ng/mL. By confocal laser scanning microscopy, the Pgp was mainly localized to the apical plasma membrane of both control and TNF alpha-treated cells. By contrast, IFN gamma induced up-regulation of both mRNA MDR1 and Pgp protein expression without incidence on Pgp activity. Interestingly, a colocalization of Pgp with lateral F actin was observed. Associated with TNF alpha, IFN gamma produced neither an antagonist nor synergistic effect on Pgp activity. In conclusion, our results demonstrate an inhibitory effect of TNF alpha and no effect of IFN gamma on Pgp transport activity using rhodamine 123 as a substrate. Mechanisms of action of these cytokines remain to be studied. PMID- 15124211 TI - Experimental design on single-time-point high-throughput microsomal stability assay. AB - An experimental design for a single-time-point microsomal stability assay was evaluated as compared with multiple-time-point studies. Results obtained from single-time-point experiments are in excellent agreement with those from multiple time points. First-order reaction kinetics revealed rapid changes of predicted half-life from percent remaining of the parent compound at the inflection points, suggesting a maximum predictive limit for half-life. Selection of the incubation time in single-time-point assays is important to obtain balanced information for stable and unstable compounds. A short incubation time (e.g., 5 min) is most useful for differentiating between unstable compounds, which is beneficial to direct the synthetic efforts in projects with poor metabolic stability. A long incubation time (e.g., 30 min) is more applicable to a compound series with high metabolic stability. For screening purposes, a moderate incubation time (e.g., 15 min) is recommended to achieve good resolution and a sufficiently high maximum predictive limit for half-life. This study suggests that a single-time-point assay is sufficient for ranking compounds in early drug discovery. It increases throughput and reduces turnaround time and cost. PMID- 15124212 TI - Rapid ESI-MS method for examining the thermal decomposition of pharmaceuticals. AB - A Finnigan MAT TSQ 700 triple quadrupole mass spectrometer equipped with a Harvard 22 syringe pump has been used to investigate the thermal decomposition of three common pharmaceuticals, acetaminophen, indomethacin, and mefenamic acid. In addition, comparative measurements on these same drugs were carried out by HPLC after thermally degrading them at elevated temperatures in order to evaluate the usefulness of the ESI-MS as a rapid means of determining the relative stability and identifying thermal decomposition products. Similar results were obtained between the two techniques (i.e., the relative rates of decomposition were mefenamic acid > indomethacin > acetaminophen), with the advantage that the ESI MS approach was much quicker to perform. PMID- 15124213 TI - Evaluation of the relationship of the molecular aggregation state of amphotericin B in medium to its genotoxic potential. AB - This work analyzes the genotoxicity potential, in the G2 phase of the cellular cycle, of an amphotericin B (AmB) commercially available form (Fungizone), and correlates it with the physicochemical properties of this product in aqueous media. The genotoxic studies were performed using peripheral blood lymphocytes from human donors. The chromosome aberrations and mitotic index were determined. Absorption spectra of Fungizone were obtained by dispersion of the stock solution in water for injection at various AmB concentrations, and using different cuvette path lengths for spectrophotometric determination. The absorption spectra of Fungizone in water are concentration dependent. High concentrations of Fungizone present a spectrum with an intense band at 340 nm, characteristic of AmB self association. Conversely, at low concentrations, the spectra are similar to those obtained with AmB in methanol, with a positive band at 409 nm, assigned to AmB monomeric form. Similarly, the cytogenetic analysis shows an important decrease on the mitotic index, which is also concentration dependent when compared with control. Furthermore, the chromosome aberrations present a small, not statistically significant, increase only at the highest concentration. The results suggest that the Fungizone presents a cytotoxicity similar to membrane pore formation in mammalian cells that depends on the existence of self associated AmB. In the presence of only monomeric forms, this phenomenon disappears. However, no genotoxicity was observed in this study. PMID- 15124214 TI - Interaction of epirubicin HCl with surfactants: effect of NaCl and glucose. AB - The interaction of an antitumoural drug, Epirubicin HCl, with anionic (sodiumdodecylsulfate; SDS), cationic (cetyltrimethylammonium bromide; CTAB), and nonionic (t-octylphenoxypolyethoxyethanol; TX-100, polyoxyethylenesorbitanmonolaurate; Tween 20) surfactants has been studied by absorption spectra as a function of surfactant concentration ranging from the premicellar to postmicellar region. At the concentrations below the critical micelle concentration (CMC), the equilibrium complex formation constant between Epirubicin cations and SDS anions has been determined by Job's method. Above the CMC, binding constant (K(b)) of Epirubicin to various types of micelles has been calculated by means of the Benesi-Hildebrand Equation. The nonionic surfactant micelles showed stronger interaction than the ionic SDS micelles, and the binding tendency of Epirubicin followed the order: Tween 20 > TX-100 > SDS. Binding of Epirubicin also has been studied in the presence of NaCl and glucose because it is administered to patients intravenously in 0.9% NaCl or 5% glucose solution. The additives have been observed to affect the CMC of the surfactants and the Epirubicin-micelle binding constant appreciably. The presence of NaCl and glucose lowered the CMC of all the surfactants studied. The binding constant of Epirubicin decreased in the presence of NaCl but increased in the presence of glucose. The equilibrium complex formation constant between Epirubicin and SDS decreased in the presence of NaCl compared with purely aqueous media. PMID- 15124215 TI - Glibenclamide transdermal patches: physicochemical, pharmacodynamic, and pharmacokinetic evaluations. AB - In the present study, matrix type transdermal patches containing glibenclamide were prepared using different ratios of ethyl cellulose (EC)/polyvinylpyrrolidone (PVP) and Eudragit RL-100 (ERL)/Eudragit RS-100 (ERS) by solvent evaporation technique. The possible drug and polymer interaction was studied by infrared spectroscopy, differential scanning calorimetry, and HPTLC analysis. All the prepared formulations were subjected to physicochemical studies (thickness, weight variation, drug content, moisture content and uptake, and flatness), in vitro release and in vitro permeation studies through mouse skin. The results suggested that there was no interaction between drug and polymers. Variations in drug release/permeation profiles among the formulations studied were observed. The microphotographs obtained by scanning electron microscopy showed the formation of pores on the surface of the patches after in vitro skin permeation studies. Based on physicochemical and in vitro skin permeation studies, the formulations with EC:PVP (3:2) and ERL:ERS (4:1) were selected for in vivo experiments. The hypoglycemic activity of the patches in comparison with oral glibenclamide administration was studied for acute (24 h) and long-term (6 weeks) effect in both normal and streptozotocin-induced diabetic mice. Various biochemical parameters (serum levels of high-density lipoprotein-cholesterol, triglycerides, total cholesterol, alanine transaminase, aspertate transaminase, urea, and creatinine and liver protein and glycogen content) and histopathological (liver, pancreas and stomach) studies were carried out in diabetic mice after treating for 6 weeks. The patches were subjected to skin irritation test (by both visual observation and histopathological evaluation), oral glucose tolerance test and pharmacokinetic evaluation in mice. The results revealed that the patches successfully prevented the severe hypoglycemia in the initial hours, which is the major side effect associated with oral route. The patches maintained similar effect during long-term treatment also. The transdermal systems produced better improvement with all the tested biochemical parameters compared to oral administration. They produced improved repair of the tissues after diabetes induced tissue injury and exhibited negligible skin irritation. The pharmacokinetic evaluation showed that the patches could maintain almost steady-state concentration of drug within the pharmacologically effective range for prolonged period of time. The better in vivo performance of the transdermal patches of glibenclamide in comparison with oral administration could be due to day-to-day glycemic control on long-term application. PMID- 15124216 TI - A new nonlinear equation for the tissue/blood partition coefficients of neutral compounds. AB - A nonlinear model equation based on tissue composition (a content of lipids, proteins, and water) for the tissue/blood partition coefficients of compounds was developed. Based on this model, our nonlinear regression analysis for neutral compounds partitioning into the kidney, brain, muscle, lung, liver, heart, and fat resulted in equations with high fitting power (training set: n = 166, r(2) = 0.851, s = 0.260, Q(2) = 0.833) and strong predictive power (test set: n = 49, r(2) = 0.851, s = 0.246, Q(2) = 0.836). This model shows that the tissue/blood partition coefficients of a compound depend strongly on tissue compositions. The magnitudes of partition coefficients of a compound in different tissues are mainly modulated by volume fractions or weight fractions of tissue compositions. PMID- 15124217 TI - Benzyl alcohol-induced destabilization of interferon-gamma: a study by hydrogen deuterium isotope exchange. AB - The destabilizing effect of a multidose preservative, benzyl alcohol, on IFN gamma was investigated. Hydrogen-deuterium isotope exchange (HX) detected by mass spectrometry (MS) was used to detect tertiary structure changes and measure global unfolding rates. The experiments showed that tertiary structure changes previously reported using circular dichroism may involve only a limited portion of the protein with the hydrophobic core of the protein remaining intact. Protein unfolding rates measured by hydrogen exchange were very sensitive to benzyl alcohol concentration, and increased markedly when salt was also added. Dynamic light scattering and size-exclusion chromatography showed that a small fraction of the protein formed large aggregates during the first few days. Measurements at longer incubation times (up to 8 days) showed that a significant fraction of protein was trapped in a structure less protected from hydrogen exchange, but not completely unfolded. This fraction of protein may be responsible for the irreversible loss of activity observed in earlier studies. PMID- 15124218 TI - Modeling the deagglomeration of micronized benzodiazepines from powder mixtures added to dissolution media. AB - The objective of this research was to model benzodiazepine deagglomeration profiles of percent agglomerated versus time when interactive mixtures containing micronized benzodiazepines were added to water. Micronized diazepam, nitrazepam, oxazepam, and, for ternary mixtures, micronized sodium lauryl sulfate were mixed with lactose-povidone granules (250-355 microm). After rapid dissolution of the lactose granules, bimodal particle size distributions of benzodiazepines, determined by laser diffraction particle sizing, represented dispersed and agglomerated distributions. The concentrations of agglomerated particle decreased with time and approached constant values. Deagglomeration profiles were determined and best modeled by a three-parameter single-exponential decay equation. A nonlinear least-squares approach was used to estimate the concentration of dispersible (C(0)) and nondispersible agglomerates (C(0a)) and the deagglomeration rate constant (K(a)). Increasing benzodiazepine and sodium lauryl sulphate concentrations in the lactose-povidone mixtures increased both dispersible and nondispersible agglomerate concentrations. Deagglomeration rate was relatively fast with half-lives around 15 min. The estimated parameters of C(0a) and K(a) may provide useful information in optimizing the design of formulations of poorly water soluble, micronized drugs to maximize their dispersion. PMID- 15124219 TI - Volume of distribution at steady state for a linear pharmacokinetic system with peripheral elimination. AB - The problem of finding the steady-state volume of distribution V(ss) for a linear pharmacokinetic system with peripheral drug elimination is considered. A commonly used equation V(ss) = (D/AUC)*MRT is applicable only for the systems with central (plasma) drug elimination. The following equation, V(ss) = (D/AUC)*MRT(int), was obtained, where AUC is the commonly calculated area under the time curve of the total drug concentration in plasma after intravenous (iv) administration of bolus drug dose, D, and MRT(int) is the intrinsic mean residence time, which is the average time the drug spends in the body (system) after entering the systemic circulation (plasma). The value of MRT(int) cannot be found from a drug plasma concentration profile after an iv bolus drug input if a peripheral drug exit occurs. The obtained equation does not contain the assumption of an immediate equilibrium of protein and tissue binding in plasma and organs, and thus incorporates the rates of all possible reactions. If drug exits the system only through central compartment (plasma) and there is an instant equilibrium between bound and unbound drug fractions in plasma, then MRT(int) becomes equal to MRT = AUMC/AUC, which is calculated using the time course of the total drug concentration in plasma after an iv bolus injection. Thus, the obtained equation coincides with the traditional one, V(ss) = (D/AUC)*MRT, if the assumptions for validity of this equation are met. Experimental methods for determining the steady-state volume of distribution and MRT(int), as well as the problem of determining whether peripheral drug elimination occurs, are considered. The equation for calculation of the tissue-plasma partition coefficient with the account of peripheral elimination is obtained. The difference between traditionally calculated V(ss) = (D/AUC)*MRT and the true value given by (D/AUC)*MRT(int) is discussed. PMID- 15124220 TI - Profile of P-glycoprotein distribution in the rat and its possible influence on the salbutamol intestinal absorption process. AB - The intrinsic absorption of salbutamol in different intestinal segments of the rat was measured and related with the corresponding intestinal P-glycoprotein (P gp) expression levels. The apparent absorption rate constants (k(a), h(-1)) observed in each fraction by means of the "in situ" rat gut absorption method after perfusion of a 0.29-mM isotonic solution of salbutamol were used as absorption indexes. In a separate series of studies, a semiquantitative analysis of the mRNA expression of P-gp by means of polymerase chain reaction and Western blot with an antibody raised against the P-gp were also performed. The "in situ" k(a) values determined in the different segments (h(-1)) showed that the absorption is not homogeneous along the intestinal tract, that is, 0.499 +/- 0.054 for colon, 0.474 +/- 0.052 for the proximal segment, 0.345 +/- 0.014 for the mean, and 0.330 +/- 0.023 for the distal fraction. Addition of verapamil to the perfusion fluid did provide a better absorption of salbutamol in the distal segment. The analysis of the mRNA expression and levels of P-gp showed that the enzyme content in each section of the intestine was inversely related to salbutamol absorption. PMID- 15124221 TI - Mg,Al layered double hydroxides with intercalated indomethacin: synthesis, characterization, and pharmacological study. AB - Magnesium aluminium layered double hydroxides (LDH) with a molar Mg/Al ratio of 2.0 have been prepared with intercalated indomethacin following two routes: reconstruction from a previously calcined Mg(2)Al-CO(3) LDH, and coprecipitation from the corresponding chlorides. The solids have been characterized by powder X ray diffraction, FTIR, and (13)C CP/MAS NMR spectroscopies and thermal stability (differential thermal analysis and thermogravimetric analysis). Intercalation of the drug is attained by both routes; however, while coprecipitation leads to a single layered structure, contamination with another layered MgAl-CO(3) phase occurs by the reconstruction method. The amount of drug intercalated, as well as the height of the gallery, are larger by the coprecipitation than by the reconstruction one. The data obtained support a somewhat tilted, upwards orientation of the drug molecules forming an interdigited bilayer, in the case of the sample prepared by coprecipitation, with the carboxylate groups pointing towards the hydroxyl layers. However, in the case of the sample prepared by reconstruction, the molecules are forming a tilted, upwards monolayer. The solids prepared are stable up to 250 degrees C. Pharmacological studies in vivo show that intercalation of the drug in the LDH reduces the ulcerating damage of the drug. PMID- 15124222 TI - Transgenic Cre expression mice for generation of erythroid-specific gene alterations. AB - Transgenic mice that express Cre recombinase in erythroid cell lineages were developed so that genes affecting erythropoiesis/hematopoiesis may be altered without necessarily affecting fetus viability. A micro-LCR cassette-beta-globin promoter-Cre recombinase gene (microLCR-betapr-Cre) construct was synthesized and used to generate transgenic mice. Concurrently, we produced mice containing a microLCR-loxP-flanked beta sickle gene (microLCR-loxP-beta(S)-loxP) construct. microLCR-betapr-Cre mice with intact transgenes in variable copy number were identified. Cre expression was assessed by RNAse protection and RT-PCR. Cre function was ascertained by breeding to microLCR-loxP-beta(S)-loxP mice. We demonstrate that beta(S) expression was not detected in the blood of bigenics, but the gene was present in nonerythroid cells. Thus, excision of the loxP flanked beta(S) gene was restricted to erythroid cell lineages. PMID- 15124223 TI - Essential roles of BMPR-IA signaling in differentiation and growth of hair follicles and in skin tumorigenesis. AB - Hair differentiation and growth are controlled by complex reciprocal signaling between epithelial and mesenchymal cells. To better understand the requirement and molecular mechanism of BMP signaling in hair follicle development, we performed genetic analyses of bone morphogenetic protein receptor 1A (BMPR-IA) function during hair follicle development by using a conditional knockout approach. The conditional mutation of Bmpr1a in ventral limb ectoderm and its derivatives (epidermis and hair follicles) resulted in a lack of hair outgrowth from the affected skin regions. Mutant hair follicles exhibited abnormal morphology and lacked hair formation and pigment deposition during anagen. The timing of the hair cycle and the proliferation of hair matrix cells were also affected in the mutant follicles. We demonstrate that signaling via epithelial BMPR-IA is required for differentiation of both hair shaft and inner root sheath from hair matrix precursor cells in anagen hair follicles but is dispensable for embryonic hair follicle induction. Surprisingly, aberrant de novo hair follicle morphogenesis together with hair matrix cell hyperplasia was observed in the absence of BMPR-IA signaling within the affected skin of adult mutants. They developed hair follicle tumors from 3 months of age, indicating that inactivation of epidermal BMPR-IA signaling can lead to hair tumor formation. Taken together, our data provide genetic evidence that BMPR-IA signaling plays critical and multiple roles in controlling cell fate decisions or maintenance, proliferation, and differentiation during hair morphogenesis and growth, and implicate Bmpr1a as a tumor suppressor in skin tumorigenesis. PMID- 15124224 TI - Drosophila eye disc margin is a center for organizing long-range planar polarity. AB - Planar polarity patterning involves long-range signaling and signal transduction. In Drosophila eye, Dishevelled (Dsh) is not only crucial for cell-autonomous transduction of a polarity signal(s) but is also involved in nonautonomous signaling function. To identify the sites for long-range polarity signaling in eye disc, we examined spatial and temporal conditions for nonautonomous Dsh function. Here we show that Dsh and its downstream factor Armadillo (Arm) are required in the border region of eye disc between the peripodial membrane (PM) and the disc proper (DP) for nonautonomous signaling. Conditional misexpression of Dsh or Arm at the posterior margin of the disc was sufficient to induce nonautonomous polarity reversals. A critical time window for the induction of such changes was approximately coincident with the timing of morphogenetic furrow initiation. Our data suggest that the disc margin is an essential site for organizing planar polarity during the initial stage of retinal morphogenesis. PMID- 15124225 TI - Inactivation of the murine Transferrin Receptor 2 gene using the Cre recombinase: loxP system. AB - Transferrin Receptor 2 (TfR2) is a key molecule involved in the regulation of iron homeostasis. Mutations in TfR2 lead to type 3 hemochromatosis in humans. We have developed mice with a targeted deletion of TfR2. The Cre-recombinase:loxP system used to create the mice allows both full deletion and tissue-specific deletion of TfR2. The development of these mice will provide new models for type 3 hemochromatosis and assist in determining the role of TfR2 in iron metabolism. PMID- 15124226 TI - The Ser(+r83k) mutation is a second site mutation of SerD affecting the N terminus of serrate. AB - The Serrate gene encodes an essential ligand for Notch signaling used during development of the adult wing and other systems in Drosophila melanogaster. Animals heterozygous or homozygous for the Ser(D) allele of this gene display characteristic defects in wing margin formation. We have characterized a spontaneously arising intragenic suppressor of Ser(D) named Ser(+r83k). Homozygous double mutant Ser(+r83k), Ser(D) animals are viable, with normal wing margin formation, but display an aberrant outspread wing posture. The two mutations can be separated by meiotic recombination which restores the Ser(D) mutant phenotype and demonstrates that in the absence of Ser(D) the Ser(+r83k) mutation is homozygous lethal. These two mutations therefore display allelic compensation. Molecular analysis reveals a single C-T transition mutation within the 5' (protein encoding region) of the Ser(+r83k) transcript. This mutation is predicted to change Arginine(176) to Cysteine, possibly leading to altered interactions with the Notch receptor. PMID- 15124227 TI - A keratin K5Cre transgenic line appropriate for tissue-specific or generalized Cre-mediated recombination. AB - We describe here a mouse line bearing a bovine keratin K5Cre recombinase transgene. These mice showed a dual pattern of Cre-mediated recombination, depending on the parent transmitting the transgene. In paternal transmission, recombination occurred specifically in the skin and stratified epithelia-as expected according to the expression of endogenous keratin K5. However, constitutive recombination between loxP sites transmitted by the sperm took place when the mother possessed the K5Cre transgene, even when the transgene was absent in the progeny. Cre expression in late-stage oocytes, with the Cre protein persisting into the developing embryo, leads to the constitutive recombination observed. Thus, this transgenic line allows for both tissue-specific and generalized recombination, depending on the breeding scheme. PMID- 15124228 TI - Laser capture microdissection of fluorescently labeled embryonic cranial neural crest cells. AB - This study is the first to report a unique genetic strategy to permanently label mammalian neural crest cells (NCC) with a fluorescent marker, selectively isolate the labeled NCC or their derivatives during murine ontogenesis by laser capture microdissection (LCM), and prepare molecular components, such as RNA, for selective gene expression analyses. Through utilization of a Cre recombinase/loxP system, a genetic strategy that has been used repeatedly to achieve tissue specific activation of reporter transgenes in mice, a novel two-component mouse model was created in which neural crest cells (and their progeny) are indelibly marked throughout the pre- and postnatal lifespan of the organism. To generate this mouse model, a Wnt1-Cre transgenic line was crossed with a mouse line expressing a conditional reporter transgene ("floxed" enhanced green fluorescent protein). Resulting offspring, expressing both the Wnt1-Cre and "floxed" EGFP alleles, demonstrated EGFP expression in the NCC and all of their derivatives throughout embryonic, postnatal, and adult stages. In the present study, EGFP labeled cranial NCC from the first branchial arch of gestational day 9.5 murine embryos were visualized in frozen tissue sections and isolated by LCM under epifluorescence optics. RNA was extracted from "captured" cells and amplified by double-stranded cDNA synthesis and in vitro transcription. Amplified mRNA samples from "captured" cells were evaluated by TaqMan quantitative, real-time PCR for the expression of a panel of NCC gene markers. The molecular genetic strategy delineated in this report will facilitate future embryo-genomic and -proteomic analyses of mammalian NCC that will serve to further our understanding of these pluripotent embryonic progenitor cells. PMID- 15124229 TI - Loss-of-imprinting of Peg1 in mouse interspecies hybrids is correlated with altered growth. AB - Previous studies have shown that loss-of-imprinting (LOI) is a regular occurrence in interspecies hybrids of the genus Peromyscus. Furthermore, evidence was presented that indicated that LOI is involved in a placental hybrid dysgenesis effect resulting in abnormal placental growth and thus possibly in speciation. We show here that LOI of the strictly paternally expressed gene Peg1 (also called Mest) occurs in F1 hybrids between Mus musculus (MMU) and M. spretus (MSP). Peg1 LOI is correlated with increased body weight and increased weight of two of the organs tested, kidney and spleen. X-gal staining of tissues derived from Peg1(+/ ) x MSP F1 mice, carrying a maternal LacZ knock-in allele of Peg1, demonstrates that LOI is stochastic in that it affects different tissues to variable extents and that, even within one tissue, not all cells are similarly affected. Furthermore, this expression from the maternal allele does not necessarily follow the endogenous paternal Peg1 expression pattern. Our results indicate that LOI occurs in interspecies hybrids in the genus Mus and that altered growth is a frequent outcome of LOI. PMID- 15124230 TI - Should vaccines and antiviral therapy for influenza now be deployed strategically? PMID- 15124231 TI - SEN virus infection. AB - The SEN virus (SEN-V) belongs to a recently discovered group of DNA viruses whose members (SEN-V-D and SEN-V-H) are associated with post-transfusion hepatitis. It is a single-stranded circular, non-enveloped DNA virus of approximately 3600 to approximately 3800 nucleotides with at least three open reading frames (ORFs). Eight different strains of SEN-V have been identified and provisionally classified as members of the Circoviridae family, a group of small, single stranded, non-enveloped circular DNA viruses that includes the TT virus (TTV), TUS01, SANBAN, PMV and YONBAN. Prevalences in different populations show great variability with marked differences between different countries and groups. Although parenteral transmission is very likely, other routes of transmission cannot be excluded. Mother to infant transmission has been demonstrated. The effect of SEN-V on chronic liver diseases has been studied. The influence of SEN V on the response to HCV therapy was investigated in three studies, with contradictory results. Data for other acute and chronic liver diseases are sparse. Further studies are needed to define the pathogenesis and clinical importance of SEN-V infection. PMID- 15124232 TI - New drugs and treatment for respiratory syncytial virus. AB - The respiratory syncytial virus (RSV) is a global health problem affecting infants and the elderly and claiming more lives than AIDS in many parts of the world. Only two antibody drugs are approved for its prevention, and ribavarin, a relatively nonspecific antiviral, is used for treatment. In the mid-1990s, a number of pharmaceutical and biotech companies initiated research programs against RSV. Together, the academic and the industrial R&D covered the whole spectrum of antibodies, vaccines, synthetic small molecule antiviral and antisense technology, and at one point, accounted for at least 25 active R&D programs. However, coincident to the marketing of the monoclonal antibody palivizumab (Synagis) in 1998, a sharp decline in such projects ensued. Many companies recently cancelled RSV projects during a prioritisation of their R&D portfolios although the continuing medical need, large market size and sales projections clearly indicate that a safe and effective RSV drug or vaccine is likely to attain blockbuster status. Today RSV receives an insignificant fraction of the R&D budget compared with AIDS, for example. This article reviews the present status of the anti-RSV regimen, covers drugs in the market and in development, and attempts to link basic research to industrial drug development, animal models of RSV, clinical trials, current clinical management, and present and future market projections. It is hoped that the unmet medical need of the victims of RSV will encourage continued involvement of the pharmaceutical and biotechnology industry in developing safe and effective prevention and treatments for RSV. PMID- 15124233 TI - A human murine mammary tumour virus-like agent is an unconvincing aetiological agent for human breast cancer. AB - There has recently been renewed interest in both the scientific literature, and in the media, that a human relative (HMLV) of murine mammary tumour virus (MMTV) may be implicated in the aetiology of up to 42% of sporadic cases of human breast cancer. Such reports are potentially of considerable clinical significance, as aside from the small percentage of genetically acquired breast cancers, the cause of sporadic cases of breast cancer is completely unknown. Indeed, convincing proof of an infectious cause for human breast malignancies would permit the development of new preventative measures, treatment modalities and raise the possibility of prophylactic and therapeutic vaccines. Here we review the historical background for a retroviral cause of human breast cancers and give an up-to-date critique of the most recent research findings. We conclude that the available evidence for an infectious aetiology of human breast cancers is unconvincing. Amongst the many cognate arguments against an infective hypothesis for sporadic cases of human breast cancer are the facts that: (i). human tissues lack the appropriate cell-surface receptor for entry of MMTVs/HMLVs; (ii). unlike all other virally caused human malignancies, immunodeficiency does not predispose to an increased incidence, or prevalence, of human breast cancers; and, (iii). reports of PCR amplification of MMTV/HMLV sequences from breast cancers have been robustly disputed by four independent laboratories. Indeed positive PCR results may be readily explained by the mis-amplification of host genomic DNA. Hence, the burden of proof currently lies with those who champion a viral cause for sporadic cases of breast cancer. PMID- 15124234 TI - JC virus genotyping offers a new paradigm in the study of human populations. AB - A small DNA virus, named JC virus (JCV) and belonging to the Polyomaviridae, is attracting the attention of anthropologists worldwide, as JCV genotyping appears to be a novel means of elucidating human migrations and the origins of various ethnic groups. The basic properties of JCV, the regional distributions of JCV genotypes, and the phylogenetic relationships among various JCV genotypes are described. Then, a study is described in which the origin of the modern Japanese was extensively investigated using the JCV genotyping method. Based on JCV genotypes in neighboring areas, the origins of people who carried JCV genotypes to the Japanese Archipelago are discussed. Finally, the relationships between JCV genotypes and Y-chromosome haplogroups are examined, as genetic variation on the Y chromosome has recently been examined in detail to investigate ancient human migrations and the population structures of human groups. PMID- 15124235 TI - BK virus-associated nephropathy in kidney transplant recipients. AB - Polyomavirus BK has emerged as an important cause of renal allograft infection leading to allograft dysfunction and loss in kidney transplant recipients. Significant progress has been made, particularly in the area of diagnostic methods for BK virus, thereby facilitating diagnosis, screening and monitoring of infection. This review outlines current concepts on the epidemiology, pathogenesis, diagnosis and therapy of BK virus nephropathy. The precise risk factors that are important for induction and progression of infection to invasive disease, the most effective diagnostic strategies, and the efficacy of current therapeutic approaches, all remain to be defined fully. It is to be hoped that these deficiencies will stimulate research to address these important questions. PMID- 15124236 TI - Abstracts of the 7th International Congress of Vertebrate Morphology. Boca Raton, Florida, USA. 27 July-1 August, 2004. PMID- 15124237 TI - Whither the ACR20? PMID- 15124238 TI - Is erythrocyte sedimentation rate the preferable measure of the acute phase response in rheumatoid arthritis? PMID- 15124239 TI - Subclinical atherosclerosis in systemic lupus erythematosus. PMID- 15124240 TI - The osteoprotegerin/osteoprotegerin ligand family: role in inflammation and bone loss. PMID- 15124241 TI - Is there anything else we could possibly need to know about COX-2 selective inhibitor drugs? PMID- 15124242 TI - Functional status and radiographic joint damage are associated with health economic outcomes in patients with rheumatoid arthritis. AB - OBJECTIVE: This analysis examines the relationship between the functional and radiographic measures of disease activity and the employment status in patients with rheumatoid arthritis (RA). We also assessed the influence of improvement in physical function on employability, healthcare costs, and quality of life, utilizing data collected in the ATTRACT trial. METHODS: During the ATTRACT trial, the Health Assessment Questionnaire (HAQ) disability index, radiographic damage measured by the van der Heijde modified Sharp (vdH-Sharp) score, employment status, healthcare resource utilization, and quality of life measured by Medical Outcomes Survey Short Form-36 were assessed at baseline and again periodically through Week 54. Clinically important improvement was defined as an improvement in the HAQ of > or = 0.25 from baseline to Week 54. RESULTS: There was a significant association at baseline between functional status and the percentages of patients employed. Increased radiographic joint damage was associated with lower full-time employment rate, with patients in the 2 highest quartiles (vdH Sharp score > 51.5) of radiographic damage having lower rates of full-time employment than those with less damage. During the ATTRACT trial, patients who achieved a clinically important improvement in HAQ scores had a significant improvement in their employability (21% vs 3%; p < 0.001), in their time lost from work (7 vs 30 days; p = 0.012), in their total/direct medical costs (7093/6791 US dollars vs 11,712/10,039 US dollars; p < 0.001), and in their quality of life (p < 0.001) compared with those who did not demonstrate this improvement. CONCLUSION: Functional disability and radiographic joint damage are correlated with employment in patients with RA. Clinically important improvement in HAQ scores is associated with substantial health economic and quality of life benefits for patients with RA. PMID- 15124243 TI - Criteria for improvement in rheumatoid arthritis: alternatives to the American College of Rheumatology 20. AB - OBJECTIVE: Appropriate outcome measures are critical to estimating treatment effects in clinical trials and observational studies. The American College of Rheumatology 20 (ACR 20) is the standard measure to assess treatment effects in rheumatoid arthritis (RA) randomized controlled clinical trials (RCT). Although the ACR 20, which measures at least a 20% improvement in a number of different measures, is a very useful dichotomous measure for identifying novel treatments with potential activity, it may fail to discriminate among active treatments. In an effort to increase stringency, trials have used the ACR at 50% and 70%. We investigated the behavior of these and other scoring systems to quantify treatment outcome in several RCT in RA. METHODS: The Cooperating Clinics for the Systematic Study of Rheumatic Disease (CSSRD) database contained 2 trials with a total of 6 arms. Using raw data for each patient entered, we calculated ACR 20, 50, and 70 for each arm. We also calculated the average number of criteria met for the ACR 20, 50, and 70 and their respective areas under the curve over time. RESULTS: The ACR 20 failed to discriminate among active therapies; however, the ACR 50 was too stringent, and only one patient in these trials satisfied the ACR 70. The average number of criteria satisfied at the 50% level at the final trial visit discriminated well, as did the area under the curve. CONCLUSION: The average number of ACR criteria met at a 20% or 50% level discriminated better than the traditional ACR criteria in these trials. More of the information is preserved by the area under the curve of the number of ACR criteria satisfied at each level because area preserves both the number of criteria and the time dependence. The area under the curve of the number of ACR criteria met is a discriminatory, specific, time-dependent, responsive, and domain-preserving metric to use as the primary outcome measure in trials of agents for the treatment of RA. These conclusions should be tested in additional data sets. PMID- 15124244 TI - Glucocorticoids and insulin sensitivity in rheumatoid arthritis. AB - OBJECTIVE: To evaluate the effects of glucocorticoids on cardiovascular (CV) risk in rheumatoid arthritis (RA). METHODS: We recorded demographic, clinical, disease outcome, and treatment variables in 92 consecutive RA patients who were not taking lipid-lowering or antidiabetic medications. Fasting blood tests were taken for determination of lipids, ultra sensitive C-reactive protein (CRP), rheumatoid factor, insulin, and glucose. Insulin sensitivity was determined using the Quantitative Insulin Sensitivity Check Index (QUICKI). RESULTS: Seventy-four (80%) patients were women, 80 Caucasian, 9 Asian, 2 of mixed ancestry and 1 Black. Their mean (95% confidence interval, CI) age, disease duration, and followup duration at our clinic were 56 (54-58), 11 (9-13) and 6 (5-6) years, respectively. Thirty-seven (40%) patients had received oral prednisone [cumulative dose 4.8 (2.0-8.5) g; duration one month to 20 years], and all patients had received pulsed (intraarticular, intramuscular, and/or intravenous) methylprednisolone [cumulative dose 2.0 (1.6-2.6) g]. Glucocorticoids were not associated with obesity, hypertension, or dyslipidemia. Having taken prednisone and high yearly frequencies of pulsed glucocorticoid administrations were associated with decreased insulin sensitivity (p < 0.05). After controlling for body mass index, ever having taken prednisone and high doses of pulsed glucocorticoids were independently associated with decreased insulin sensitivity (p < 0.05). CONCLUSION: Previous exposure to oral prednisone and high doses of pulsed glucocorticoids were associated with decreased insulin sensitivity in RA. Since decreased insulin sensitivity is an independent risk factor for CV disease, glucocorticoids may contribute to the excess CV event rates in RA. PMID- 15124245 TI - Increase of cyclooxygenase-2 expression by interleukin 15 in rheumatoid synoviocytes. AB - OBJECTIVE: To determine the effect of interleukin 15 (IL-15) on cyclooxygenase-2 (COX-2) expression in rheumatoid synoviocytes. METHODS: Fibroblast-like synoviocytes (FLS) were prepared from the synovial tissues of patients with rheumatoid arthritis (RA) and cultured in the presence of IL-15. Levels of COX-2 mRNA and protein expression were determined by reverse transcription-polymerase chain reaction and Western blot, respectively. ELISA was used to measure concentrations of IL-1beta, tumor necrosis factor-a (TNF-a), and prostaglandin E2 (PGE2) in the culture supernatants. RESULTS: IL-15 dose-dependently increased COX 2 mRNA and protein expression in FLS, but not the COX-1 mRNA level. Both IL-1beta and TNF-a upregulated COX-2 mRNA comparably to IL-15, but neither IL-2 nor interferon-g had any effect on the COX-2 mRNA level. Treatment with anti-IL-1beta or anti-TNF-a antibodies partially reduced the IL-15-stimulated COX-2 mRNA expression, suggesting that these cytokines may take part in modulating COX-2 by IL-15. Dexamethasone and pyrolidine dithiocarbamate, but not curcumin, completely blocked the IL-15-induced upregulation of COX-2 mRNA. A gel mobility shift assay revealed that nuclear factor-kB (NF-kB) was one of the major signal molecules to mediate IL-15-induced COX-2 upregulation. The increase of COX-2 by IL-15 is PGE2 dependent because exogenous PGE2 reversed the suppressive effect of NS-398, a selective COX-2 inhibitor, on COX-2 mRNA and protein expression. CONCLUSION: This study confirms the effect of IL-15 on upregulation of COX-2 in a PGE2-dependent manner. The activation of NF-kB bound to the COX-2 promoter appears to be a downstream target of IL-15 stimulation in FLS, exerted either directly or through the increase in IL-1beta and TNF-a production. PMID- 15124246 TI - Relative sensitivity to change of the erythrocyte sedimentation rate and serum C reactive protein concentration in rheumatoid arthritis. AB - OBJECTIVE: To compare the sensitivity to change of the erythrocyte sedimentation rate (ESR) and serum C-reactive protein (CRP) concentration used as measures of rheumatoid arthritis (RA) activity. METHODS: A literature search was conducted to identify all clinical trials and observational studies of disease-modifying medications and corticosteroids in RA that reported results for both ESR and CRP before treatment and 4 weeks to 24 weeks after treatment in the same patients. For each test, effect sizes were computed as the change in the test with treatment divided by the pretreatment standard deviation. A pooled analysis was performed on the paired differences in effect sizes for ESR and CRP within each study. RESULTS: One hundred twenty-three studies with 184 active treatment arms were identified that included measurements of both ESR and CRP. Sixty-three studies with 90 active treatment arms provided sufficient data to permit calculation of effect sizes, and were included in the analysis. In the 36 treatment arms that reported results at 12 weeks, the ESR was more sensitive to change than the CRP, with a paired difference in effect sizes of 0.09 units (95% confidence interval 0.03, 0.15; p = 0.005). In the 76 treatment arms that reported results at 24 weeks, the ESR was also more sensitive to change than the CRP, with a paired difference in effect sizes of 0.11 units (95% CI 0.05, 0.17; p = 0.0004). CONCLUSION: In these studies of disease-modifying medications in RA, the ESR was more sensitive to change than the CRP at 12 weeks and 24 weeks of treatment. Few studies examined changes in these measures at times earlier than 12 weeks. PMID- 15124247 TI - Radiation synovectomy of the ankle with 75 MBq colloidal 186rhenium-sulfide: effect, leakage, and radiation considerations. AB - OBJECTIVE: In a retrospective study we evaluated the effect, duration of effect, and safety of radiosynoviorthesis of the ankle in patients with persistent synovitis, refractory to disease modifying antirheumatic drugs (DMARD) and intraarticular glucocorticoid injections. We estimated leakage and dose to target and non-target organs. METHODS: Radiation synovectomy was performed by injection of 75 MBq 186rhenium colloid and 20 mg triamcinolone-hexacetonide mixed in a volume of about 1.5 ml. About 24 hours after injection, leakage of the radionuclide was measured with a single-head gamma camera, with views of the ankle joint, regional (inguinal) lymph nodes, and liver. Leakage was expressed as counts in the target region of interest corrected for background relative to total counts corresponding with percentage of injected dose. The effect of radiosynoviorthesis was scored into 3 categories: (1) No effect, i.e., persistent synovitis or only minimal reduction of swelling and/or pain, or the need of intraarticular glucocorticoid injection within 3 months or arthrodesis of the treated joint within 6 months. (2) Moderate effect, i.e., significant reduction of swelling, pain, and improvement of function. (3) Good effect, i.e., complete or almost complete remission of synovitis. RESULTS: The mean age of patients (28 women, 12 men) at the time of treatment was 58 years (range 33-76); 54 consecutive procedures in ankles of the 40 patients were evaluated. No effect was found in 12 of 54 (22%) treated joints; moderate effect in 12 (22%), with a mean duration of effect of 34 months (range 12-49); and good effect in 30 (56%), with a mean duration of effect of 41 months (range 21-75). Mean effect-duration did not differ significantly between the moderate and good effect groups. Mean leakage did not differ significantly between the effect groups. CONCLUSION: Radiation synovectomy of the ankle is a safe and effective treatment in persistent synovitis, although all patients eventually experienced recurrence of arthritis. PMID- 15124248 TI - Economic evaluation of folate supplementation during methotrexate treatment in rheumatoid arthritis. AB - OBJECTIVE: To determine cost-effectiveness of folic or folinic acid supplementation in patients with rheumatoid arthritis (RA) who started methotrexate (MTX) treatment. METHODS: An economic evaluation, performed alongside a randomized, double blind, placebo controlled trial with followup of 48 weeks. Patients started MTX with placebo (n = 137), folic acid (n = 133), or folinic acid (n = 141). Outcome measures were drug survival and quality-adjusted life-years (QALY), measured with the EuroQol questionnaire. Both medical and nonmedical costs were analyzed. RESULTS: Drug survival after 48 weeks was 60% for placebo, 81% for folic acid, and 87% for folinic acid. QALY during a 48 week period were 0.55 (95% CI 0.52-0.58) in the placebo group, 0.55 (95% CI 0.52-0.58) in the folic acid group, and 0.58 (95% CI 0.56-0.60) in the folinic acid group. Mean medical costs were 1398 US dollars (placebo), 1409 US dollars (folic acid), and 1776 US dollars (folinic acid). Mean total costs were 3339 US dollars, 3632 US dollars, and 3296 US dollars, respectively. CONCLUSION: In terms of resource deployment, no statistically significant difference was found between the 3 strategies. The preferred strategy consists of folic acid supplementation because of improved drug survival. PMID- 15124249 TI - Duplex study of the carotid and femoral arteries of patients with systemic lupus erythematosus: a controlled study. AB - OBJECTIVE: To identify atherosclerosis in the common carotid (CCA) and common femoral arteries (CFA) of patients with systemic lupus erythematosus (SLE) and matched controls. METHODS: Fifty-one consecutive patients with SLE were enrolled in the study. Controls were matched by age, sex, ethnicity, and atherosclerosis risk factors. All patients and controls underwent ultrasonic biopsy (U-B) of the CFA and CCA, a noninvasive screening technique that detects early atherosclerotic plaques and changes. The U-B features were classified and scored as follows: class A: normal (score 0); class B: interface disruption (score 2); class C: intima-media granulation (score 4); class D: plaque without hemodynamic disturbance (score 6); class E: stenotic plaque (score 8); and class F: plaque with symptoms (score 10). Total score was calculated. Classes A and B indicate an intact media; classes D to F point to a significant medial involvement; class C signifies a borderline lesion with a potential for regression to normal or progression to a plaque. RESULTS: Mean ages were 40.5 years for SLE patients and 41 years for controls (p = 0.6). Ninety-six percent of the patients and controls were women. The mean disease duration of SLE was 8.65 years. Frequencies of risk factors among the SLE patients compared to controls were hypertension (30% vs 24%), smoking (23% vs 24%), and dyslipidemia (17.7% vs 17%). No patient had diabetes mellitus or family history of cardiovascular disease. A 3.17-fold increased rate of atherosclerotic plaques was detected in the SLE patients compared with controls (95% CI 1.08-10.9). Twenty-eight percent of SLE patients had at least a single class D-F lesion in one of the 4 vessels tested, compared with 10% in the control group (p = 0.02). In addition, the mean total U-B score of the SLE patients was significantly higher than that of the controls (5.65 vs 3.14; p = 0.02). Univariate analyses showed that the development of plaques in SLE was associated with a history of ischemic heart disease, hypertension, cardiovascular accident, and anemia. Multivariate analysis found plaques to be strongly associated with age, particularly in those older than 50 (OR 2.66, p = 0.000). CONCLUSION: Patients with SLE have a high rate of atherosclerotic changes compared to controls. The development of atherosclerosis is strongly associated with age. PMID- 15124250 TI - The effect of race on disease activity in systemic lupus erythematosus. AB - OBJECTIVE: To determine and contrast the disease activity and clinical variables between Hispanic and Caucasian patients with systemic lupus erythematosus (SLE) in New Mexico. METHODS: Socioeconomic-demographic and clinical data were collected from 125 SLE patients by an interview-administered questionnaire. The Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) was used to assess disease activity. RESULTS: Seventy-four Hispanics (H) and 40 Caucasians (C) were compared. Demographics including age, gender, disease duration, marital status, and cigarette smoking were similar between the 2 groups. However, education and income were higher in the Caucasian group compared to the Hispanic group. There was no significant difference between the 2 groups in overall disease activity as measured by the SLEDAI. However, when individual components of the SLEDAI were compared, Hispanics had an increased prevalence of arthritis (77% vs 51%, p = 0.01) and depressed complement levels (40% vs 18%, p = 0.02). Moreover, corticosteroid use was higher among the Hispanics compared to the Caucasian population (p = 0.03). CONCLUSION: We found similar levels of overall disease activity in Hispanic and Caucasian patients with SLE. However, Hispanics used more corticosteroids, had a greater prevalence of arthritis, and had depressed complement levels indicating increased SLE disease activity highly restricted to specific domains. It remains to be determined whether these restricted but discrete differences are genetic in origin, or are related cultural or environmental factors. PMID- 15124251 TI - Antibodies to tissue transglutaminase and Saccharomyces cerevisiae in ankylosing spondylitis and psoriatic arthritis. AB - OBJECTIVE: Subclinical gut inflammation has been described in patients with ankylosing spondylitis (AS) or psoriatic arthritis (PsA). Joint involvement has also been reported related to celiac disease. We investigated IgA antibodies to bovine tissue tranglutaminase (tTg) and IgA and IgG antibodies to human tTg and to Saccharomyces cerevisiae (ASCA) in patients with AS and PsA. METHODS: We evaluated the frequency of IgA antibodies to bovine tTg, and of IgA and IgG antibodies to human tTg and to ASCA in 43 patients with AS and 75 with PsA. As control groups we considered 79 patients with rheumatoid arthritis (RA) and 78 healthy blood donors. RESULTS: We detected antibodies as follows: IgA antibodies to bovine tTg in 1/43 patients with AS, 3/75 with PsA, 1/79 with RA, and in 9/78 healthy controls; IgA antibodies to human tTg in 1/43 patients with AS, 1/75 with PsA, 1/79 with RA, and in 3/78 healthy controls; IgG antibodies to human tTg in 1/43 patients with AS, 4/75 with PsA, 5/79 with RA, and in 7/78 healthy controls. IgA ASCA were confirmed in 10/43 patients with AS, 7/75 with PsA, 14/79 with RA, and in 7/78 healthy controls; IgG ASCA were present in 5/43 patients with AS, 4/75 with PsA, 8/79 with RA, and in 8/78 healthy controls. No statistically significant difference was observed in the prevalence of IgA or IgG antibodies to bovine and human tTg and in the frequency and in mean level of IgA or IgG ASCA between the studied groups or between each group and healthy controls. CONCLUSION: Our data fail to show an increased prevalence of autoantibodies associated with celiac and Crohn's disease in patients with AS and PsA. PMID- 15124252 TI - Unique angiopathy after herpes virus infection. AB - OBJECTIVE: We describe 3 Japanese patients with peculiar renal and/or coronary arterial stenosis and/or multiple aneurysms after herpes virus infection, following ischemic symptoms. We investigated for viral antigens and viral DNA in situ, and for shared abnormalities of cellular immunity. METHODS: Panarteriography was performed diagnostically, and patients were grouped as follows: 3 patients with peculiar renal and/or coronary artery narrowing and/or multiple aneurysms; another 3 patients with renal fibromuscular dysplasia; and other young adults with effort angina, with no history of herpes virus infection, as controls. Detection of viral antigens and viral DNA in situ was done by polymerase chain reaction method and immunohistochemical staining. Cellular immunity was examined at the time of ischemic symptoms. RESULTS: Viral antigens and DNA were scarcely detected, except in herpes zoster skin lesion with leukocytoclastic vasculitis. However, shared abnormalities of cellular immunity, such as a decreased CD4+ T cell number and reduced natural killer cell activity, were more prominent in the 3 patients with unique vasculopathy after herpes virus infection. CONCLUSION: Unique vasculopathy following herpes virus infection might be a more severe and extensive disease. We speculate that sustained viral infection, repetitive activation of virus related antigens, and suppressed immune state might contribute to formation of peculiar vascular alterations. PMID- 15124253 TI - Development of the BD-QoL: a quality of life measure specific to Behcet's disease. AB - OBJECTIVE: Current outcome measures for patients with Behcet's disease (BD) are impairment-focused and do not necessarily take account of the wider impact of the condition on the individual's lifestyle. Our aim was to develop a disease specific measure of quality of life (QoL) for BD. METHODS: The content of the BD QoL was derived from qualitative interviews with patients using a "needs-based" approach to identify items. A postal survey was used to test the scaling properties, reliability, internal consistency, and validity of the new questionnaire using Rasch analysis. A second postal survey was used to assess test-retest reliability and internal consistency and to provide further evidence of the validity of the questionnaire. RESULTS: Main themes emerging from the qualitative interviews included relationships, emotions, limitations in day to day activities, and self-image. From these themes 71 statements were chosen as potential items for the BD-QoL. After analysis, 30 items of the BD-QoL emerged free of item bias for age and sex. Fit to the Rasch model was excellent. In the second postal survey test-retest reliability of the 30 item BD-QoL was 0.84. CONCLUSION: The BD-QoL provides the clinician with a simple, reliable, and valid tool for assessing the influence of interventions for BD and for evaluating models of service delivery. It is well accepted by patients, and has excellent scaling and psychometric properties. The BD-QoL complements information obtained through BD-specific disease activity scales. PMID- 15124254 TI - Resource utilization in postmenopausal osteoporosis without incident fractures. AB - OBJECTIVE: To evaluate the annual utilization and cost incurred by postmenopausal women with osteoporosis in a public tertiary healthcare facility. METHODS: One hundred consecutive patients with postmenopausal osteoporosis were selected from the Universidade Federal de Sao Paulo (UNIFESP), Brazil. The socioeconomic and clinical characteristics, quality of life, and resource utilization and costs in the last year were assessed using 3 questionnaires. Each resource used had its unitary cost based on the Public Healthcare System Table of Fees (1998). RESULTS: The mean age was 66 years and the monthly family income was 456 US dollars (1998). There was a mean of 7 visits/patient/year. Seventy-seven percent of the patients used calcium and 38% used estrogens. There were no incident fractures during the study. The mean total annual cost for osteoporosis treatment, considering the societal perspective, was 775 US dollars (1998) per patient/year. CONCLUSION: The costs of all treatment drugs related and not related to osteoporosis represented 9% of the monthly household income. Among some drugs used, many are lacking evidence of effectiveness, such as shark cartilage. The costs related to transportation were significant, representing 21% of the total directly spent by the patients. Further studies are necessary to rationally drive treatment, based on the best cost-effectiveness strategies of osteoporosis management. PMID- 15124255 TI - Immunogenicity of hylan g-f 20 in Guinea pigs and mice. AB - OBJECTIVE: To determine the product-specific immunogenicity of a chemically modified sodium hyaluronate derivative, hylan G-F 20, that is used in the treatment of osteoarthritis of the knee. METHODS: Guinea pigs were subcutaneously immunized with hylan G-F 20 (Synvisc) once a week for 3 weeks. After resting, these animals received an intradermal challenge with hylan to elicit allergic skin reactions. Animal sera were tested for the presence of hylan-specific antibodies by homologous passive cutaneous anaphylaxis (PCA) assay and of anti hylan IgG by ELISA. Further, mice were similarly immunized with hylan, and their sera were analyzed by heterologous PCA assay in rats and by ELISA for anti-hylan Ig(G+M) and anti-hylan IgE. RESULTS: In the guinea pig studies, acute and delayed erythematous skin reactions were elicited in immunized animals after the intradermal challenge with hylan. The sera of hylan-immunized guinea pigs showed positive reaction in the homologous PCA assay and significantly high amount of anti-hylan IgG, whereas the sera did not show any cross-reactivity against sodium hyaluronate. Hylan also exhibited immunogenicity in mice of 3 inbred strains, and C3H/HeN mice showed higher production of anti-hylan antibodies than Balb/c and C57BL/6 mice. CONCLUSION: Hylan G-F 20 exhibited immunogenicity in guinea pigs and mice. Recent reported severe acute inflammatory reactions in human patients after repeated intraarticular injections of hylan G-F 20 might involve product specific, immune-mediated mechanisms. PMID- 15124256 TI - A population based historical cohort study of the mortality associated with nabumetone, Arthrotec, diclofenac, and naproxen. AB - OBJECTIVE: To identify the unbiased differences in all cause mortality among populations using 4 non-steroidal antiinflammatory drugs (NSAID): nabumetone, Arthrotec, diclofenac plus a cytoprotective agent dispensed separately (diclofenac+), and naproxen. METHODS: We performed a population based historical cohort study using linked data from several provincial health care databases. Logistic regression was used to produce estimates of the mortality associated with the study drugs unbiased by known confounders. The entire population of the province of Saskatchewan, Canada entitled to drug plan benefits in 1995 was eligible (approximately 91% of 1 million people). Participants were identified if they filled a prescription for one of the 4 study NSAID (18,424 individuals). They were then followed forward in time for 6 months to determine all cause mortality. RESULTS: Compared to nabumetone, the adjusted odds of death for participants taking Arthrotec was 1.4 (95% confidence interval, CI: 0.9-2.1), for diclofenac+ 2.0 (1.3-3.1), and naproxen 3.0 (1.9-4.6). CONCLUSION: The multivariate analysis showed patients taking nabumetone and Arthrotec had significantly lower mortality than those taking other study drugs. Nabumetone had 1/3 to 1/5 the mortality associated with the diclofenac+ and naproxen groups. It appears that inherent gastroprotective strategies in the study NSAID may translate into decreased mortality at the population level. PMID- 15124257 TI - Intraarticular osteoid osteoma: clinical features, imaging results, and comparison with extraarticular localization. AB - OBJECTIVE: Intraarticular osteoid osteoma is uncommon and presents diagnostic difficulties, which are important for both rheumatologists and orthopedic surgeons. Clinical symptoms, imaging procedures, differential diagnostic problems, and treatment results of intraarticular as compared with extraarticular osteoma are analyzed in this retrospective study. METHODS: Nineteen patients with intraarticular osteoid osteomas (Group A), with a mean followup period of 34 months, are compared with 15 others with extraarticular lesions (Group B). RESULTS: Nine intraarticular tumors were located in the hip, 3 in the elbow, 6 in the ankle, and one in the first metatarsal head. The nonspecific symptoms in Group A, such as chronic synovitis, decreased range of motion, joint effusion, contractures, and lack of the intense perifocal sclerotic margin on radiographs, led to significant delay in diagnosis (on average 26.6 mo in Group A, 8.5 mo in Group B). The extreme variety of previous diagnoses at referral reflect the problems of differential diagnosis. A detectable nidus is often absent on conventional radiograph. Bone scintigraphy is unspecific and often fails to visualize the nidus. Computed tomography scans were accurate in two-thirds of the intraarticular and in 90% of extraarticular cases. Magnetic resonance image findings, although sometimes controversial, provided essential additional information for the correct diagnosis and therapy. CONCLUSION: Clinical symptoms and imaging signs of intraarticular osteoid osteomas were significantly different from the classical hallmarks of extraarticular lesions. The 10% intraarticular occurrence of osteoid osteomas in this series is not as rare as some investigators suggest. The radiological and clinical findings are uncharacteristic and misleading, and the lesions are difficult to identify. Careful search for history data, such as nocturnal pain and positive salicylate test, in addition to extensive imaging procedures, led to the correct diagnosis prior to surgery in two-thirds of our patients with intraarticular osteoid osteomas. PMID- 15124258 TI - Rheumatic disease in an Australian Aboriginal community in North Queensland, Australia. A WHO-ILAR COPCORD survey. AB - OBJECTIVE: To estimate prevalences of rheumatic diseases in Aboriginal Australians. METHODS: The methodology of the Community Oriented Program for the Control of Rheumatic Diseases (COPCORD) was followed. Everyone aged 15 years or older in Yarrabah, North Queensland, was invited to complete a COPCORD Core Questionnaire. Aboriginal health workers carried out a house-to-house survey during January 2002. People reporting current musculoskeletal symptoms and 56 others (controls) were examined at the community health center. RESULTS: Eighty percent of the target population was covered during the survey. Eight hundred and forty-seven questionnaires were completed (47% men) and 135 people refused, a response of 86%. Rheumatic symptoms within the previous 7 days were reported by 33% and past symptoms by 22%. The most common sites of current pain were low back (12.5%), knee (11.2%), and shoulder (8.9%). Sixty-seven people (7.7%) said activities were limited by their symptoms. Two hundred and sixty-three people were examined, and the most common diagnoses were soft tissue pain (point prevalence 7.4%), osteoarthritis (5.5%), and low back pain (4.3%). The cumulative prevalence of gout was 7.0% in men and 0.9% in women over the age of 15 years. The relative risk of gout associated with drinking regularly was 2.5, and with body mass index > 25 was 3.3. No rheumatoid arthritis or systemic lupus erythematosus cases were identified, but there were 4 cases of psoriatic arthritis (point prevalence 0.5%). CONCLUSION: This is the first unselected population study of rheumatic diseases in Australian Aboriginals. There was a high prevalence of gout among men, with modifiable factors of weight and alcohol identified. PMID- 15124259 TI - Significant variation exists in home care services following total joint arthroplasty. AB - OBJECTIVE: To determine a preliminary profile of the variation in rehabilitation and home care services for patients with total joint arthroplasty (TJA) in Ontario in 2001. METHODS: A cross-sectional survey was conducted of directors at the 43 regional community care access centers (CCAC). RESULTS: One-third (36%) of CCAC had existing care pathways, 54% had defined discharge criteria, and 32% had predetermined the length of home care services. The intensity and frequency of services provided were variable. CONCLUSION: There is a need to standardize rehabilitation protocols to maintain quality of care and contain costs. PMID- 15124260 TI - Depressed mood impedes pain treatment response in patients with fibromyalgia. AB - OBJECTIVE: To investigate prognostic factors in the course of the fibromyalgia syndrome (FM) from baseline to post-treatment. METHODS: Fifty-seven patients with FM were examined in a randomized intervention study. Pre-treatment variables were entered into linear regression analyses: gender, age, duration of disease, allocation to treatment, pain distribution (based on a patient-made drawing), fatigue, sleep disturbance, and depressed mood (based on visual analog scores), with pain distribution at treatment completion as the dependent variable. RESULTS: Depressed mood at baseline was a significant predictor of sustained widespread pain at treatment completion. CONCLUSION: The findings indicate a role for depressed mood as a predictive factor for treatment response. PMID- 15124261 TI - Assessment of osteoporosis by quantitative ultrasound versus dual energy X-ray absorptiometry in children with chronic rheumatic diseases. AB - OBJECTIVE: To evaluate the validity of quantitative ultrasound bone sonometry (QUBS) as a screening tool for the diagnosis of osteoporosis in children with chronic rheumatic diseases (CRD), compared to the conventional dual energy x-ray absorptiometry (DEXA). METHODS: Forty children with CRD [32 with juvenile idiopathic arthritis (JIA), 6 with systemic lupus erythematosus, and 2 with dermatomyositis] aged 9.9 +/- 4.3 years, were evaluated by QUBS of radius and tibia and DEXA of the lumbar spine. Twenty-five (62.5%) patients were treated with corticosteroids. Measurements of the velocity of the ultrasound wave, expressed as speed of sound (SOS) in m/s, and the results of the bone mineral density (BMD) assessed by DEXA were compared to reference data from healthy age and sex matched Israeli children. RESULTS: Compared to controls, patients with CRD had significantly lower values by QUBS and DEXA alike. BMD and SOS z scores < -1 SD were found in 45% and 38% of the patients, respectively. Reduced BMD and SOS values correlated with age at disease onset and corticosteroid treatment. BMD alone correlated negatively with disease duration and methotrexate therapy. BMD was significantly lower in patients with polyarticular JIA compared to patients with oligoarticular disease (p < 0.03). SOS values did not differ between subtypes of JIA. A significant positive correlation was found between the lumbar DEXA and radius SOS. CONCLUSION: QUBS evaluation of radius and tibia yielded results comparable to DEXA and may therefore be used for screening patients with CRD for osteoporosis. QUBS might represent a promising means of evaluating bone quality in at-risk children. PMID- 15124262 TI - Osteoprotegerin (OPG)/RANK-L system in juvenile idiopathic arthritis: is there a potential modulating role for OPG/RANK-L in bone injury? AB - OBJECTIVE: To evaluate serum levels of osteoprotegerin (OPG) and receptor activator of nuclear factor kB-ligand (RANK-L) in patients with juvenile idiopathic arthritis (JIA); to correlate these values with disease activity variables, radiological bone damage, and bone mass; and to correlate OPG gene polymorphisms with bone mass. METHODS: Eighty-four patients (66 girls and 18 boys) with JIA and 40 sex and age-matched controls were enrolled. Serum OPG and RANK-L were measured using an enzyme-linked immunosorbent assay. OPG genotyping was performed by polymerase chain reaction. RESULTS: Patients with JIA had significantly higher levels of serum OPG than controls (p = 0.001) and lower levels of RANK-L in comparison with controls (p = 0.0003). The OPG/RANK-L ratio in patients was higher than in controls (p = 0.004). No significant correlations were found between disease duration, erythrocyte sedimentation rate, and C reactive protein values with either OPG or RANK-L serum levels. A significant difference in serum OPG levels (but not in RANK-L) was found between patients with and without erosions (p = 0.008). No correlation was found between OPG and RANK-L levels and bone mass (DXA Z scores). A higher prevalence of OPG CC genotype was found in both patients (65.4%) and controls (82.5%) (p = 0.006). Subjects with CC genotype had a higher lumbar spine bone mineral density (LS BMD). CONCLUSION: We evaluated for the first time levels of OPG and RANK-L in children with JIA. The higher OPG/RANK-L ratio in JIA might be the result of a compensatory production of OPG. The presence of the T allele of the OPG gene appears to be associated with low BMD. PMID- 15124263 TI - Nitric oxide-derived species in synovial fluid from patients with juvenile idiopathic arthritis. AB - OBJECTIVE: To evaluate superoxide anion (O2-), nitrite/nitrate (NO2-/NO3-), and nitrotyrosine (NT) production and the contribution of myeloperoxidase (MPO) to the production of NT-containing proteins in the synovial fluid (SF) of patients with juvenile idiopathic arthritis (JIA). The affected tissues in inflammatory arthritis produce large amounts of nitric oxide (NO) or peroxynitrite (ONOO-) but there are no reports of NO or ONOO- participation in JIA. We also attempted to correlate our findings with variables of disease activity and articular damage. METHODS: We analyzed 40 patients with JIA, mean age 12.7 years, mean disease duration 7.8 years. O2- production was measured by cytochrome C reduction after incubation of 106 synovial fluid (SF) cells with or without phorbol myristate acetate (PMA), formyl-methionyl-leucyl-phenylalanine (FMLP) or opsonized zymosan. SF and serum NO2-/NO3- levels were measured by Griess reaction; NT was detected by Western blot. Myeloperoxidase (MPO) activity was estimated spectrophotometrically. Clinical and laboratory variables [erythrocyte sedimentation rate, C reactive protein (CRP), and radiological score] and interleukin 6 (IL-6) levels were evaluated. RESULTS: NO2-/NO3- production was greatly enhanced in the joints of JIA patients (54.6 +/- 3.2 micro M) when compared with serum (13.9 +/- 0.6 micro M; p < 0.001). NO2-/NO3- levels in SF were positively correlated with the number of infiltrating lymphomononuclear cells. NT-modified proteins detected in the SF showed a high correlation with radiological score, disease duration, CRP, and IL-6. CONCLUSION: Our results confirm the increased oxidative stress in children with JIA, suggesting a high in situ production of NO. The positive correlation between the expression of NT modified proteins and variables of disease activity and damage is additional evidence that nitrogen and oxygen species may be involved in the joint destruction seen in patients with JIA. PMID- 15124264 TI - Another patient with chromosome 18 deletion syndrome and juvenile rheumatoid arthritis. AB - Previously, 4 children with deletion of the long arm of chromosome 18 and chronic arthritis were reported. We present an 8-year-old girl with arthritis, atrial septal defect, external auditory canal atresia, and developmental delay. She is the fifth child reported with 18q- syndrome and juvenile rheumatoid arthritis. Evidence is mounting that genetic loci on chromosome 18 may play a role in the expression of complex autoimmune diseases. Idiopathic arthritis should be considered as a potential additional feature in 18q- syndrome. PMID- 15124265 TI - Embolic complications of a mitral valve rheumatoid nodule. AB - We describe a patient with a rheumatoid nodule on the mitral valve who developed embolic phenomena from an overlying thrombus. It is important to recognize that thrombus can develop on intracardiac rheumatoid nodules and that these patients may require anticoagulation. PMID- 15124266 TI - Three-dimensional power Doppler sonographic visualization of synovial angiogenesis in rheumatoid arthritis. PMID- 15124267 TI - The fate of abstracts: without publication, science is dead. PMID- 15124269 TI - Serious gastrointestinal events from low dose analgesic use. PMID- 15124270 TI - Serious gastrointestinal events from low dose analgesic use. PMID- 15124272 TI - Ophthalmologic monitoring for antimalarial toxicity. PMID- 15124273 TI - Diagnostic value of anticyclic citrullinated peptide antibody in rheumatoid arthritis. PMID- 15124276 TI - Prevalence of silent amyloidosis in RA and its clinical significance. PMID- 15124278 TI - Macrophage activation syndrome due to methotrexate in a 12-year-old boy with dermatomyositis. PMID- 15124280 TI - Breast implant related disease. PMID- 15124282 TI - Hepatitis C virus and rheumatoid arthritis: further pieces to the puzzle. PMID- 15124283 TI - Etanercept in breast milk. PMID- 15124284 TI - Severe recurrent neurological disease in the MAGIC syndrome. PMID- 15124285 TI - [Vaccinations in pig health care in the Netherlands--analysis of a questionnaire among veterinarians]. AB - Veterinarians specializing in pig health care responded to a questionnaire regarding their experiences with vaccinations. The goal of the questionnaire was to gain insight into a) the reasons for vaccination and b) the factors involved in the efficacy of vaccination. The results indicated that vaccinations were typically initiated because of health problems at the farm. Veterinarians worked together with farmers and other concerned parties to initiate vaccinations. Respondents predicted that the number of vaccinations would decrease substantially with optimal farm management, but would increase if farmers were allowed (under strict conditions) to vaccinate their own pigs. The results further indicated that the nature of financial compensation predicted the rate of vaccination. Veterinarians paid according to a fixed 'fee for service' system vaccinated less frequently than did veterinarians who were paid according to an ongoing farm management contract. In conclusion, veterinarians appear to be restricted in their capacity to disseminate their experience and knowledge due to the competing needs of the farming network (animals, farmers, retailers). PMID- 15124286 TI - [There are more dogs called Rover]. PMID- 15124287 TI - [The General Food Law]. PMID- 15124288 TI - [Reflection on the KKM (milk quality) and PBB (periodic visitations): are there perspectives?]. PMID- 15124289 TI - [Idiopathic gastric rupture in a Friesian foal]. AB - Acute idiopathic gastric rupture resulted in sudden death of a 4.5-months-old healthy Friesian foal. No morphological lesions were present which could explain the preceding gastric impaction. In this case hypodypsia may have been (partially) responsible for the impaction as only a mechanical drinking apparatus, unsuitably for horses, was available. PMID- 15124290 TI - [Political attention for animal welfare and health]. PMID- 15124291 TI - [BVD management: how do you get infected farms virus-free?]. PMID- 15124292 TI - [Reaction from the Dutch Food Industry Companion Animals to the letter sent titled 'welfare claim']. PMID- 15124293 TI - [Update of business employers' negotiations]. PMID- 15124294 TI - Try these claims-appeal strategies. PMID- 15124295 TI - The English patient's EMR. PMID- 15124296 TI - How to discuss end-of-life care. Here's a guide to making advance care planning a part of your practice. PMID- 15124297 TI - What is an FP? Specialty leaders went through an extensive survey process last year. Here's a preliminary look at their findings. PMID- 15124298 TI - I thought I understood breast cancer. PMID- 15124299 TI - Why won't they help my patient? PMID- 15124300 TI - Buying a densitometer. FAQs answered. PMID- 15124301 TI - The separate worlds of the physician-defendant. PMID- 15124302 TI - Why make time for behavioral counseling? PMID- 15124303 TI - Is your call coverage working? PMID- 15124304 TI - Extra hospital days can cost you plenty. PMID- 15124305 TI - Group practice. The best way to divide income. PMID- 15124306 TI - Keeping tabs on insurance plans. PMID- 15124307 TI - A knowledge base fit for all midwives. PMID- 15124308 TI - More on cesareans. PMID- 15124309 TI - More on cesareans. PMID- 15124310 TI - Marion's message. Support nurse-midwives. PMID- 15124311 TI - What do you consider essential midwifery knowledge? What do you consider advanced midwifery knowledge? Who owns that knowledge? PMID- 15124312 TI - Ways of knowing. Open and closed systems. PMID- 15124313 TI - Wild, beautiful birth. PMID- 15124314 TI - Women's innate knowledge. PMID- 15124315 TI - Passing it on. PMID- 15124316 TI - The knowing body and remembering heart. PMID- 15124317 TI - An intimate dance with birthing women. PMID- 15124318 TI - Ideas on writing protocols for your practice. PMID- 15124319 TI - Omega 3 oils and pregnancy. AB - Women can safely commence omega 3 intake in early pregnancy to allow the full benefits to be incorporated into the body. Pregnant women should consider the intake of omega 3 oils and evening primrose oil throughout pregnancy in order possibly to prevent preterm delivery, promote an easier birth, assist the baby's brain and eye health and preprogram the baby's cell membranes for optimum lifelong wellness. Benefits to the mother may include prevention of postnatal depression and the comfort of knowing that she is giving her baby a healthy start in life. PMID- 15124320 TI - Woman's work. PMID- 15124321 TI - The business of midwifery. An introduction. PMID- 15124322 TI - Herbal friend for birthing mothers: partridgeberry. PMID- 15124323 TI - Interviews with maiden midwives. Reaching out to the next generation. Interview by Mary Kroeger. PMID- 15124324 TI - Nursing the caesarean born. PMID- 15124325 TI - Midwifery and chiropractic. Bonding for life. PMID- 15124326 TI - The birth of a collaboration. PMID- 15124327 TI - Making birth humane in Brazil. PMID- 15124328 TI - Suspect diagnoses come with biophysical profiling.. PMID- 15124329 TI - More on cesareans. PMID- 15124330 TI - Nuchal cord response. PMID- 15124331 TI - Growing against the grain. PMID- 15124332 TI - Rx for investors. PMID- 15124333 TI - Children's dental health month: a different perspective. PMID- 15124334 TI - Breaking up doesn't have to be hard to do. PMID- 15124335 TI - New York City agency seeks volunteer emergency responders. PMID- 15124336 TI - Medicaid children. A vulnerable cohort. AB - Vulnerable populations face special problems in dealing with the complexities of our fragmented health care delivery system, creating impediments to timely and effective care. Persistent differences in health status, utilization and outcomes exist among racial/ethnic groups, among age groups and between the sexes. Children are one specific cohort that serves as an unfortunate example of vulnerability within this system. This paper reviews some of the barriers that exist in access to care for Medicaid children. PMID- 15124337 TI - Get ready! The infants are coming. AB - The scourge of early childhood caries is well known. Knowing when and how to approach the littliest patients and their parents still requires education and effort, but, when done correctly, leads to a winning situation for all involved. An overview of the why's and how's of incorporating pediatric patients into the general dental practice. PMID- 15124338 TI - Dental disease control in Pine Hill, New Mexico. AB - One-year results of a community-operated dental disease control project in Pine Hill, New Mexico. The program uses fluoride, chiefly rinse, and has not only reduced the amount of decay in permanent teeth, but has markedly reduced the need for restorative care of primary teeth. PMID- 15124339 TI - B. T. Whitney. Physician, dentist, inventor, entrepreneur and dedicated figther for dentistry and dental education. AB - In the mid-1850s, Western New York had become one of the most highly industrialized areas of the nation, largely because it was the terminus of the Erie Canal. Buffalo was among the foremost cities in the country; and its dentists were actively establishing a professional association. What was lacking, however, was a dental school to serve the whole area, including the territory to the west. Four leading men in the profession, who also started one of America's greatest and most innovative dental manufacturing companies, struggled to achieve that goal. Foremost among them was Benajah Ticknor Whitney, a physician, who had received his M.D. at the first medical school west of the Hudson River. But his interest in dentistry developed early in his training, and Dr. Whitney went on to become one of the state's outstanding dental leaders. Sadly, he didn't live to see his dream fulfilled. It would take 20 more years--and a hard fight to overcome opposition within the profession--until a university-affiliated school was established in Buffalo. PMID- 15124340 TI - We are not bankers. PMID- 15124341 TI - Simple blood test may provide clues to periodontal disease. PMID- 15124342 TI - Pediatrics academy calls for elimination of soda in schools. PMID- 15124343 TI - Cassetteless digital x-ray systems. AB - Cassetteless digital x-ray (DX) systems combine film-quality images and fast patient throughput. Because they are expensive, they need to be versatile enough to perform most or all radiographic projections. To meet this challenge, DX manufacturers have developed a variety of imaging detectors and gantry configurations. This presents facilities with an array of choices. In this Evaluation, we examine cassetteless DX systems from six suppliers: Canon, Fujifilm, Imaging Dynamics, Kodak, Siemens, and Swissray. We compare them with the three systems--from Del Medical, GE, and Philips--that we evaluated in our November 2001 issue. Our testing examines the ability of these systems to provide at least the same amount of diagnostic information as conventional screen-film systems while significantly increasing the overall efficiency of the radiology department. Each system provides a slightly different trade-off between image quality and usability. We rate three systems Preferred due to their excellent image quality, good ease of use, and the wide range of exams they can provide using a single detector. The remaining six devices are all rated Acceptable, although one of them is ranked slightly superior to the other five. PMID- 15124344 TI - Cassette versus cassetteless DX. PMID- 15124345 TI - A high-pressure situation: conventional i.v. sets can burst when used with CT contrast media injectors. PMID- 15124346 TI - Defective call cords used with Rauland-Borg nurse call system prevent staff notification. PMID- 15124347 TI - Philips Heartstream defibrillator interferes with ultrasound system. PMID- 15124348 TI - [The similarities and the differences of embryogenesis and carcinogenesis]. AB - The main similarities and embryonic and tumor cells, as well as the mechanisms preventing the malignant transformation of embryonic cells, are presented in this review. Special attention is paid to the role of specific polypeptide growth factors in reciprocally excluding processes: embryogenesis and carcinogenesis. Based on the presented analysis, new potential targets for antitumor drugs are considered. PMID- 15124349 TI - [Specific features of development of the male generative organs in clones of the Siberian stone pine (Pinus sibirica De Tour) of different origins in the Krasnoiarsk forest-steppe]. AB - The development of male generative organs of the Siberian stone pine clones of different origin was analyzed in the Krasnoyarsk forest-steppe for many years. Geographical and individual variability was described as concerns the number of microstrobils and pollen variability. The beginning of "flowering", abundance of microstrobils, and their regular formation were shown to markedly vary and, all other conditions being equal, be determined by the presence of clones, their hereditary features, and environmental conditions, as well by the influence of stock. PMID- 15124350 TI - [Patterns of protein biosynthesis in the embryo and endosperm during embryogenesis in Pinus sylvestris L]. AB - The patterns of protein biosynthesis in the embryo and endosperm during embryogenesis in the Scots pine were studied using electrophoresis and biochemistry methods. Proteins of the albumin-globulin fraction were visualized already at the early embryonic stages. The main polypeptide components (48-60, 37 39, and 20-22 kDa) were gradually accumulated in the course of maturation. A high synchrony was noted between the stages of embryogenesis and molecular events related to protein biosynthesis and accumulation in the developing seed. PMID- 15124351 TI - [The use of human fibroblasts grown on microcarriers for the formation of the connective tissue equivalent]. AB - Live equivalents of tissues, specifically those produced on the basis of fibroblasts and collagen gel, are widely used for repair of organ and tissues defects. In clinical practice, it is more convenient to use the fibroblasts grown on microcarriers or such a connective tissue equivalent when the fibroblasts on microcarriers are embedded in collagen gel. We studied the properties of a connective tissue equivalent produced by embedding the fibroblasts grown on microcarriers in collagen gel for its prospective use in clinical practice. According to our results, the optimal time of use of the live tissue equivalent amounts to three--four days after embedding of fibroblasts on microcarriers in gel. At that time, contraction only begins, which facilitates manipulations with the gel. PMID- 15124352 TI - [A population of neurons expressing tyrosine hydroxylase and migrating from olfactory placode into forebrain during ontogenesis in rats]. AB - Olfactory placodes, that give rise to the olfactory and respiratory epithelia during ontogenesis, are a source of many neurons migrating into forebrain in the direction of growth of the olfactory nerves. The neurons expressing gonadotropin releasing hormone (GnRH) are among the best studied in the population in question. This hormone is responsible for the central regulation of reproduction in adult animals. It was already shown that, in addition to the GnRH immunoreactive neurons, a small amount of neurons expressing tyrosine hydroxylase (TH), the first enzyme of catecholamine synthesis, migrates into the forebrain. Such a transient population of TH-immunoreactive neurons was shown by means of single and double immmunohistochemical labeling. The TH neurons were first found on branches of the olfactory, terminal, and vomeronasal nerves, along the trajectory of migration of GnRH-immunoreactive neurons on day 15 of embryogenesis, which preceded the appearance of GnRH-immunoreactive neurons. On days 17-21 of embryogenesis, both populations of neurons were found in almost the same areas and on day 21 single neurons contained both GnRH and TH. There were no neurons expressing decarboxylase of aromatic acids (DAA), the second enzyme of catecholamine synthesis, among TH-immunoreactive neurons, thus suggesting noncatecholaminergic nature of these neurons. However, single nonenzymatic DAA immunoreactive neurons were found in the area of anterior olfactory nuclei in the forebrain, which suggests their involvement in local cooperative synthesis of catecholamines in the area where GnRH-immunoreactive neurons penetrate in the forebrain. Thus, the neurons expressing TH, TH and GnRH, and DAA were found in rats during prenatal period in the nasal part of the head along the nerves projecting into the forebrain and in the rostral part of forebrain. The origin and functional significance of these neurons are discussed. PMID- 15124353 TI - [NADPH-diaphorase-positive nerve cells in heterotopic spinal cord transplants]. AB - The method of ectopic transplantation of embryonic CNS rudiments makes it possible to study the mechanisms underlying adaptation of the transplanted embryonic rudiments. The production of nitric oxide by cells is considered as one of such mechanisms. NADPH-diaphorase is an index of the presence of nitric oxide synthase in cells. It was shown that the nerve cells of rat embryonic spinal cord transplants preserved their capacity to express NADPH-diaphorase after transplantation in the sciatic nerve of an adult animal for six months. The dynamics of NADPH-diaphorase-positive neurons of rat embryonic spinal cord developing after transplantation and in situ were studied. In spinal cord neck region, small bipolar NADPH-diaphorase-positive neurons were visualized on day 17 of prenatal development. After transplantation of the embryonic (day 15) spinal cord in the nerve, NADPH-diaphorase-positive neurons were formed later than in situ: within seven days. The results of histochemical studies carried out within six months after the operation suggest a protective role of NADPH-diaphorase in the neurons of allotransplants developing under the conditions of altered microenvironment and insufficient innervation and also suggest that nitric oxide can cause the death of neurons in long surviving transplants. PMID- 15124354 TI - [The studies of temporal and spatial characteristics of somitogenesis in fish embryos]. AB - New data were obtained corroborating that somitogenesis is a rhythmic process, in which the time of somite formation is strictly constant. This constant (tau s) can be considered as a natural unit of developmental "biological clock" characterizing rhythmic processes. The constant tau s can be determined with an exceptional accuracy that has no analogs among the known biological processes. This fact alone suggests that the accuracy of developmental clock is very high. In addition to the constancy of tau s, all forming somites have equal linear size along the notochord axis. In the process with strictly constant temporal and spatial factors, time plays the leading role in triggering the formation of new somite. This became clear in studies of twin embryos. Both embryos had the same number of somites but they were shorter than in the normal embryos. Also, we measured the length of head and both segmented and unsegmented caudal parts of the trunk. Combined with the published data on somitogenesis, our results suggest that the previously proposed scheme for the role of developmental clock in embryogenesis predicts: (1) a possible loss of some embryonic stages without serious consequences for subsequent development and (2) periodic switching on/off any embryonic processes (at the molecular, cellular, or supercellular level) with intervals multiple to tau s. PMID- 15124355 TI - [Dorsoventral differences of morphogenetic potencies of the loach blastoderm in experiments with alteration of the mass of explanted fragments]. AB - We studied the influence of doubling the mass of explanted fragments of the dorsal and ventral loach blastoderm at the early gastrula stage on their capacity for differentiation of axial structures. The dorsoventral differences are as follows: the differentiation of somites correlates, according to the results of factor analysis, with the shape complication only in double dorsal explants, while the notochord is more differentiated in the ventral fragments, if it is present, than in the dorsal ones. Doubling of the mass of dorsal fragments of the blastoderm enhances their morphogenetic potencies and shifts differentiation towards the formation of trunk axial structures. The increased mass of ventral fragments does not affect their differentiation and morphogenesis, but disturbs the correlation of these processes. PMID- 15124356 TI - [Phyllotaxis types and patterns of their realization]. AB - The types of phyllotaxis have been considered. The types of phyllotaxis occurring in plants allow them to keep unchanged the position of leaves on the shoot during growth, despite unequal stretching of stem in length and diameter. PMID- 15124357 TI - Time for a leadership tune-up? PMID- 15124358 TI - Proceed with vision power. AB - Today's successful nurse managers implement a visionary leadership style. PMID- 15124359 TI - Supply and demand: it's your business. AB - Economics can help nurse managers analyze situations in which people must make choices from limited options. PMID- 15124360 TI - What's new for 2002? AB - The Joint Commission's anticipated changes for 2002 include clarifications to existing standards in three manuals and several new initiatives. PMID- 15124361 TI - Psychiatric crises and emergency admissions. AB - Emphasize safe practice to avoid legal liability when evaluating individuals in psychiatric crisis. Test your knowledge with the following questions, then check your answers at http://www.nursingmanagement.com. PMID- 15124362 TI - Stemming the tide of pleural effusions. AB - Teach staff to determine the cause and type of pleural effusion to ensure appropriate patient care. PMID- 15124363 TI - Telemetry systems help nurses work smarter. AB - Higher patient acuities on medical/surgical floors prompt use of new telemetric capabilities for bedside oxygenation monitors. PMID- 15124364 TI - Safer medication management. AB - The five rights of medication administration alone don't ensure solid medication management. A systems approach helps to prevent errors. PMID- 15124365 TI - "Techno-marvels" just around the corner. AB - In the next 18 months, cutting-edge information technology will likely become commonplace in hospitals. PMID- 15124366 TI - IC advances decrease patient and staff risk. AB - New products, research, practices, and job roles continue to advance infection control practices. PMID- 15124367 TI - I.v. risk reducers. AB - The trend of rapid advancements in infusion therapy continues, specifically regarding catheters and insertion techniques. PMID- 15124368 TI - The ABCs of PDAs. AB - The popularity of personal digital assistants (PDAs) continues to rise, due mostly to the convenience and reliability they offer caregivers. PMID- 15124369 TI - What's new at the point of care? AB - Preview the latest in near-patient specimen analysis technology, including blood glucose and occult blood tests, and ways to measure heparin's anti-coagulation. PMID- 15124370 TI - JCAHO and HCFA issue new restraint guidelines. AB - Incorporate policies and procedures into your facility's care plan that enhance safe restraint and seclusion. PMID- 15124371 TI - Breathing easy. AB - Patient care models increasingly call for collaborative initiatives between nursing and respiratory care practitioners. PMID- 15124372 TI - Self-directed work teams: use with caution. PMID- 15124373 TI - Medicare drugs. AB - The following overview knits together a collection of articles that focus on prescription drug issues as they relate to the Medicare Program. In general, the articles examine the following themes: drug cost management; drug cost estimation, and racial disparities in drug coverage and use. These commentaries provide information on private-sector experience in administering a drug benefit, models that might be used to estimate take-up rates and their associated costs, and several indepth looks at racial disparities by various chronic conditions. PMID- 15124374 TI - Prescription drug benefits: cost management issues for Medicare. AB - Little attention has been devoted in policy circles as to how Medicare would manage an outpatient prescription drug benefit. This article, first, discusses the role of the pharmacy benefits manager (PBM), the entity that processes claims and otherwise helps administer the benefit. It then discusses the major decisions that will be necessary regarding such matters as: which drugs should be covered; how broad should the pharmacy network be; whether there should be incentives to obtain generic rather than brand-name drugs when available; for drugs with no generic equivalent, should there be incentives to obtain less expensive, medically appropriate brand-name drugs; and how should prescription drug utilization be managed. PMID- 15124375 TI - Reporting of drug expenditures in the MCBS. AB - Comparing data from both the 1999 MCBS and drug utilization data supplied by the survey respondents' pharmacies, the author details the methods used to determine the level of misreporting of drug expenditures in the MCBS. Findings suggest that prescription drug expenditures are underreported by 17 percent and the number of prescriptions used is underreported by 17.7 percent. The data also identify demographic factors that predict a beneficiary's likelihood to either overreport or underreport his or her medications, as well as the extent to which beneficiaries misreport their drug use and spending. PMID- 15124376 TI - Predictability of prescription drug expenditures for Medicare beneficiaries. AB - MCBS data are used to analyze the predictability of drug expenditures by Medicare beneficiaries. Predictors include demographic characteristics and measures of health status, the majority derived using CMS' diagnosis cost group/hierarchical condition category (DCG/HCC) risk-adjustment methodology. In prospective models, demographic variables explained 5 percent of the variation in drug expenditures. Adding health status measures raised this figure between 10 and 24 percent of the variation depending on the model configuration. Adding lagged drug expenditures more than doubled predictive power to 55 percent. These results are discussed in the context of forecasting, and risk adjustment for the proposed new Medicare drug benefit. PMID- 15124377 TI - Participation and crowd-out in a Medicare drug benefit: simulation estimates. AB - This article provides information on likely participation in the Medicare prescription drug plan and expected crowd-out. We use a microsimulation model based on data from the MCBS to estimate the costs and benefits of a Medicare drug plan, including the benefits from reductions in risk. The simulations are repeated using different combinations of benefits and subsidies. In addition, the simulations explore the effects of different behavioral parameters for moral hazard (the extent to which participants increase drug spending in response to reduced costs) and risk aversion (the extent to which participants would be willing to pay to avoid risk) to identify the impact of these factors on participation and crowd-out. PMID- 15124378 TI - Racial and ethnic disparities in prescription coverage and medication use. AB - This study compared drug coverage and prescription drug use by race and Hispanic ethnicity for Medicare beneficiaries with three chronic conditions: diabetes, hypertension, or heart disease. We found that among beneficiaries without any drug coverage black persons and Hispanics used 10 to 40 percent fewer medications, on average, than white persons with the same illness, and spent up to 60 percent less in total drug costs. Having drug coverage somewhat lessened these differences although the effect was consistent with only M + C prescription benefits. Substantially lower medication use remained for dually eligible black beneficiaries and Hispanics with employer-sponsored drug benefits. PMID- 15124379 TI - Racial disparities in prescription drug use among dually eligible beneficiaries. AB - Well-documented racial disparities in use of medical services raise concerns about such disparities in other aspects of health care. We compare the difference in Medicaid pharmacy use between black and white dually eligible Medicare beneficiaries. Controlling for the presence of chronic illnesses, we find that black beneficiaries have significantly fewer prescriptions filled and lower pharmacy costs in 8 of the 10 States examined, despite having higher physician costs. If this disparity stems from a lack of provider or beneficiary knowledge, programs to educate providers or beneficiaries may hold the greatest promise for reducing it, whether pharmacy coverage is obtained from Medicaid or from a new Medicare benefit. PMID- 15124380 TI - Medicare beneficiary's use of prescription drug discount cards, CY 2002. PMID- 15124381 TI - It's all about patient care. PMID- 15124383 TI - Financial responsibility for medical care of family members--tidbits from the Iowa courts. PMID- 15124385 TI - Refusing service to "no pay" patients. PMID- 15124384 TI - Physicians rally at state Capitol for common sense tort reform. PMID- 15124386 TI - [Occupational medicine in nuclear industry and power engineering]. AB - The author analysed results of medical service in atomic industry and power engineering over 50 years. Those results are beneficial for management in occupational medicine for any new complicated and potentially dangerous technology and activity. PMID- 15124387 TI - [Occupational diseases in nuclear industry workers]. AB - The authors analysed contemporary work conditions, occupational morbidity and traumatism in atomic industry. Major types of the occupational diseases are listed. The article contains measures on lower morbidity and prophylaxis of occupational diseases and traumatism among atomic industry workers. PMID- 15124388 TI - [Results and prospects of radiation neurology]. AB - The article represents main results and prospective goals on a problem concerning neurologic disorders caused by ionizing radiation in atomic industry workers. PMID- 15124389 TI - [Medical and psychological aspects of safety measures maintenance among nuclear and power station personnel]. AB - The article deals with 15-year experience of medical and psychophysiologic service in Medical and Sanitary Establishment No. 156 and Balakovo nuclear power station on providing reliability of occupational activities for the station personnel. PMID- 15124390 TI - [Psychological services in the professional training of the NES (nuclear energy station) operators]. AB - Psychophysiologic service could play an important role in optimal training and anti-wreek practice with various tasks that simulate some aspects of operations' activities in contingency. PMID- 15124391 TI - [Clinical and functional tests for pulmonary hypertension and cor pulmonale in chronic pulmonary berylliosis and plutonium-induced pulmonary sclerosis]. AB - Berylliosis and plutonium pneumosclerosis patients appeared to have two types of pulmonary hypertension: hypervolemic and hypertonic ones. Hypervolemic type of pulmonary circulation was revealed in 31.7% of berylliosis patients and in 53.8% of plutonium pneumosclerosis patients. Hypertonic type of pulmonary circulation was seen in 68.3% of berylliosis patients and in 46.2% of plutonium pneumosclerosis patients mainly at stages II and III. Berylliosis and plutonium pneumosclerosis patients are subjects for early pulmonary hypertension--moderate in most patients at stage I and high at stage II and III. Pulmonary hypertension is a main pathogenetic factor underlying chronic corpulmonale formation. PMID- 15124392 TI - [Hemopoietic response to americium and plutonium exposure in nuclear industry workers]. AB - The examinees demonstrated definite effects caused by total influence of Am-241 and Pu-239: chronic radiation sickness with diffuse pneumosclerosis, chronic toxic radiation bronchitis and peripheral blood changes (reactive alterations and stable lower platelets and reticulocytes count), microfocal hypoplasia, osteodysplasia, reactive hepatopathy. Frequency of these effects increased with higher radiation exposure and did not depend on the age. PMID- 15124393 TI - [Medical and biological consequences of human's chronic exposure to radiation]. AB - The authors considered medical consequences of radiation accidents (burial of radioactive waste into Techa river and accident in 1957) in "Majak" Industrial Association. Results of long-term observations helped to evaluate health state of people who underwent chronic exposure to radiation and of their descendants. PMID- 15124394 TI - [Case of chronic radiation sickness with subsequent leukemia in track driver subjected to long-external gamma-irradiation (cesium-137)]. AB - The article deals with consequences seen in track driver after prolonged exposure to radiation caused by "lost" gamma-source (cesium-137) that long remained in receptacle of the vehicle's left door. Radiation dose averaged 8 Gy. The authors presented clinical manifestations, changes in peripheral blood and bone marrow, cytogenetic data by progression of radiation hemopoiesis hypoplasy to myelodysplastic syndrome and to acute leukaemia. PMID- 15124395 TI - [Acute local radiation lesions and their consequences (contingency in flaw inspection)]. AB - Illustrated with case history (patient F), the authors represented diagnosis, prognosis, treatment, long-term consequences of local radiation lesions. PMID- 15124397 TI - [Respiratory system peculiarities in workers who underwent acute toxic exposure to beryllium]. PMID- 15124396 TI - [Evaluating the significance of some factors that influence radioactive iodine intake in acute radiation sickness patients victims of Chernobyl power station accident]. PMID- 15124398 TI - Children as organ donors: a persistent ethical issue. PMID- 15124399 TI - Cognitive development and pediatric consent to organ donation. PMID- 15124400 TI - Child organ donation, family autonomy, and intimate attachments. PMID- 15124401 TI - The child as living donor: parental consent and child assent. PMID- 15124402 TI - Designing an ethical policy for bone marrow donation by minors and others lacking capacity. PMID- 15124403 TI - The child as organ and tissue donor: discussions in the Danish Council of Ethics. PMID- 15124404 TI - Children and organ donation: some cautionary remarks. PMID- 15124405 TI - CQ sources/bibliography. PMID- 15124406 TI - Response to "Utilitarianism shot down by its own men" by Tuija Takala (CQ Vol 12, No 4). Takala shoots herself in the foot. PMID- 15124407 TI - If you have said A, you must also say B: is this always true? PMID- 15124408 TI - Five ways to kill the biotech industry (and one to help it prosper). PMID- 15124409 TI - Predatory hospital billing: dynamic cost shifting to the uninsured. PMID- 15124410 TI - The HIPAA headache. It just won't go away. An exclusive survey of privacy officers says full compliance with the 1-year-old regulations is still elusive. AB - After a year of HIPAA, there's still some adjusting to do. Some don't grasp the value of accounting to patients about disclosures, but HHS' Richard Campanelli, left, says it offers patients reassurance that their data was safeguarded. Baylor's Donna Bowers, on the cover, says caregivers are now understanding that "Maybe (patients) do care that I'm asking all these personal questions in front of all these people." PMID- 15124411 TI - Concierge critique. Warning doesn't worry docs at boutique practices. PMID- 15124412 TI - More docs disciplined. PMID- 15124413 TI - Moving money. Survey may cause regional shifts in Medicare dollars. PMID- 15124414 TI - Shock to the system. Problems may force VA to forsake computer project. PMID- 15124415 TI - Wrangling in Texas. Four Parkland trustees resign after dispute with CEO. PMID- 15124416 TI - Everybody's doing it. Pending Anthem deal opens door for more mergers. PMID- 15124417 TI - The health plan challenge. Watching to see if industry rhetoric on quality, costs is backed by action. PMID- 15124418 TI - Scaling the cliff. Post-acute providers report healthier numbers despite Medicare challenges. PMID- 15124419 TI - A brand new bag. Trauma-care revolution among benefits touted as blood substitute enters critical phase of clinical trials. PMID- 15124429 TI - Is personal choice an absolute right? PMID- 15124430 TI - Reflections on a research journey. PMID- 15124431 TI - A web of connections. PMID- 15124432 TI - Community development in an African village. PMID- 15124433 TI - CNPS: respecting your privacy rights. PMID- 15124434 TI - All in the family. Interview by Barbara Sibbald. PMID- 15124435 TI - Pilot crashes. Lawsuit by four N.J. hospitals shuts down experimental gain sharing program designed to reward docs for controlling costs. AB - When Robert Wood Johnson Health System challenged a CMS gain-sharing demonstration project, its complaint was that it was excluded. But now a judge has shut down the experiment. The ruling could have ripple effects on other hospitals seeking creative ways to enlist doctors in containing costs. Hunterdon Medical Center had already recruited 75 physicians, says Robert Pickoff, left, chief medical officer. PMID- 15124436 TI - Try, try again. GOP pushes to clamp down on Medicaid abuse. PMID- 15124437 TI - Under scrutiny in N.C. $414 million in hospital payments questioned. PMID- 15124438 TI - Price-fixing probe. FTC investigates three N.M. hospitals and their IPAs. PMID- 15124439 TI - Changing the subject. Tenet names six new CMOs to reinforce patient safety. PMID- 15124440 TI - Compensation coalition. Groups seek new claims process for immigrant care. PMID- 15124441 TI - Rough and unready. Billions have been spent on terror preparedness, without much result. PMID- 15124442 TI - The nurse of the future. Fewer workers, new technologies and older patients place RNs at center stage. PMID- 15124443 TI - Front lines to front office. Nurses draw on patient-care experience, interdepartmental knowledge as they graduate to top executive positions. PMID- 15124445 TI - The field of health law: its past and future. Introduction. PMID- 15124444 TI - By the numbers. Nurse-staffing levels. PMID- 15124447 TI - Health care law and policy: whence and whither? PMID- 15124446 TI - Feng Shui and the restructuring of the hospital corporation: a call for change in the face of the medical error epidemic. PMID- 15124448 TI - Bioethics? The law and biomedical advance. PMID- 15124450 TI - A tribute to mentors. PMID- 15124449 TI - From the doctor to the system: the new demands of health law. PMID- 15124451 TI - Where is the "there" in health law? Can it become a coherent field? PMID- 15124452 TI - I've seen enough! My life and times in health care law and policy. PMID- 15124453 TI - Five easy pieces: motifs of health law. PMID- 15124454 TI - Comparative and international health law. PMID- 15124455 TI - Reflections on race and bioethics in the United States. PMID- 15124456 TI - The four ages of health law. PMID- 15124457 TI - Health law at fifty years: a look back. PMID- 15124458 TI - The growth of health law and bioethics. PMID- 15124459 TI - Where is health law going?: follow the money. PMID- 15124460 TI - A medical ghost in the e-health machine. PMID- 15124461 TI - Some reflections on teaching law and medicine in law school since the '60s. PMID- 15124462 TI - Letter to the editors of Health Matrix. PMID- 15124463 TI - [Simultaneous operations during laparoscopic cholecystectomy]. AB - Simultant laparoscopic operations were performed in 1993-2003 yrs period in 321 patients, including 287--using laparoscopic and 34--the combined (laparoscopic and open) access. Concurrent diseases were diagnosed preoperatively in 219 (68.2%) of patients and were disclosed while doing intraoperative revision--in 102 (31.8%). The simultant operations performance, as a rule, enhanced mildly the total duration of a basic stage, did not influence the duration of postoperative period and the patients rehabilitation essentially, as well as for frequency of the intra--and postoperative complications occurrence. The authors consider that it is expedient to perform laparoscopic intervention simultaneously for concurrent surgical diseases of abdominal cavity. PMID- 15124464 TI - [Technical aspects of laparoscopic cholecystectomy in umbilical hernia]. AB - There were analyzed the results of laparoscopic cholecystectomy performance in 1230 patients, in 60 (4.7%) of whom--with simultant intervention for concurrent umbilical hernia. The expediency of simultant operation performance was caused by exclusion of the concurrent disease progressing, the reduction of the temporary disablement duration and by the shortage of the treatment expenses. The management of postoperative period is similar both in simultant treatment and in conventional performance of laparoscopic cholecystectomy. PMID- 15124465 TI - [Morphological characteristics of cystic pancreatic tumors]. AB - The results of 86 clinical observations of cystic pancreatic tumor are adduced. The expediency of the postoperative pathomorphological investigation conduction was substantiated because of high degree of the cystic pancreatic tumors polymorphism presence and low informativity of preoperative and intraoperative investigations. PMID- 15124466 TI - [Application of octreotide in patients with acute pancreatitis]. AB - In 1986-2002 yrs period 515 patients were treated in the clinic for an acute pancreatitis. The severity of the disease was assessed according to clinical and laboratory data, basing on Ranson scale. Severe acute pancreatitis was diagnosed in 45 (8.7%) of patients. In 16 patients together with conventional treatment octreotide was applied in 100-300 mkg/day dosage during 5 days. Fast improvement of the patients state and elimination of acute pancreatitis symptoms was noted. All the patients are alive. PMID- 15124467 TI - [Strategies in the surgery choice in patients with hepato-pancreatic-duodenal biliary diseases involving cardiovascular and respiratory system diseases]. AB - Results of treatment of 38 patients, suffering nontumoral and tumoral diseases of the hepatopancreatoduodenal zone organs and concurrent diseases of cardiovascular and respiratory systems were analyzed. Septic-purulent affections, caused by the principal disease, had constituted the leading complication in occlusion-stenotic affection of cardiac coronary vessels. Due to the alternative absence the majority of the patients were operated on with lethal outcome. Basing on analysis of morphometric investigations of cardiac vessels there was suggested the expedience of coronarographic investigation conduction and, basing on its results, to perform preoperatively the balloon dilatation or stenting of cardiac coronary vessels, independently of kind of the main disease present. In presence of haemodynamically significant stenosis of the cardiac coronary vessels trunk and main branches (more than 70% of their lumen) the lethal outcome risk after performance of operative intervention is very high. PMID- 15124468 TI - [Experience of laparoscopic hernioplasty in the treatment of inguinal hernia]. AB - For 2001-2003 yrs period laparoscopic prosthesis hernioplasty was performed in 127 patients. Oblique reducible inguinoscrotal hernia had constituted the main nosological form of the disease. Peculiarities of preoperative preparation, anesthesiological support, operative procedures, complications postoperative management and medicinal securing of patients, immediate and late follow-up results of their surgical treatment were lighted on. The advantages of laparoscopic prosthesis hernioplasty were proved, such as the recurrence frequency lowering, social-economic efficacy, associated with reduction of the patients stationary stay, their labour rehabilitation and the recurrence absence. PMID- 15124469 TI - [Membrane plasmapheresis in complex treatment of endogenous intoxication in diffuse peritonitis of appendicular genesis in children]. AB - With the aim to lower the degree of endogenous intoxication in 9 children with diffuse peritonitis of appendicular genesis the membrane plasmapheresis with the plasma exchange was used in complex of postoperative treatment. Depending on severity of clinical course of the disease there were conducted two procedures on the 2-3 and 5-7 postoperative day to every patient. Application of the membrane plasmapheresis in complex of treatment had promoted the reduction of the endogenous intoxication degree due to influence on its main pathogenetic chains and the immune systemic reactivity raising. PMID- 15124470 TI - [Cardiac valve prosthesis in patients with infective endocarditis and chronic renal insufficiency in terminal stage on a programmable hemodialysis]. AB - Mechanical cardiac valve implantation was performed in 3 patients with infective endocarditis, sepsis, chronical renal insufficiency of terminal stage, to whom the programmed dialysis was conducted. The operation was performed for vital indications. All the patients after the operation are alive, their treatment was continued, using programmed hemodialysis, effect of operation is good. The success of treatment of infective endocarditis in patients with chronical renal insufficiency of terminal stage depends on timely radical elimination of the infection foci in endocardium, the intracardial and systemic hemodynamics normalization, application of etiotropic antibacterial therapy, an adequate programmed hemodialysis. PMID- 15124471 TI - [Experience of surgical treatment of primary peripheral pulmonary cancer and of tumoral metastases by video-assisted thoracoscopy]. AB - In clinic of thoracic surgery were operated on 2108 patients for various forms of pulmonary cancer, 218 (10.3%) of them--using videothoracoscopy (VTHS). In 141 (64.7%) patients the method of VTHS was applied for diagnostical purposes and in 55 (25.2%)--for treatment. Histological investigations of excised specimen showed planocellular cancer in 38 patients, adenocarcinoma--in 9, nonsmall cell carcinoma--in 6, anaplastic cancer--in 2. The operation cost while applying VTHS had constituted at average 648.65 US dollars, of thoracotomy--1283.5 US dollars. PMID- 15124472 TI - [Aspects of oxygen regimen and metabolism in patients after operative intervention under neuroleptic analgesia]. AB - Result of microsurgical correction of posttraumatic defect of soft tissues depends a lot on anesthesiological support. During the operation the inhibition of energetic status causes the occurrence of oxygentransporting debt, metabolic acidosis develops. Analgosedation, peculiar to anesthesy, promotes an organism defense toward operative trauma of middle severity, causes temporary hypometabolic effect, changed than by oxidantic stress and general adaptive syndrome development. PMID- 15124473 TI - [Analysis of postoperative recurrence of malignant tumors of adrenal glands]. AB - Malignant tumors of suprarenal glands (MTSG) are characterized by aggressive course, inclination to recurrences and early metastatic spreading. Causes of the recurrence occurrence of MTSG were analyzed in 292 patients. There was revealed more frequent recurrence of tumor of the adrenal gland cortical substance than of medullary substance. In anaplastic and low differentiated forms of cancer the prognosis is poor, efficacy of treatment is extremely low. Most frequently (in 63.5% of observations) the recurrence is localized in regional collectors of the lymph outflow. Elaboration of operative interventions, which include prophylactic systemic dissection of retroperitoneal fat, would have to permit to lower the recurrence frequency after primary surgical intervention. Occurrence of the tumor recurrence after primary operation constitute a bad prognostic sign. Of the patients with the tumor recurrence 55.8% are considered incurable. PMID- 15124474 TI - [Synchronous two-team surgeries of cancer and cicatricial stenosis of the esophagus]. AB - The experience of two-brigade operations concerning a cancer of the esophagus and its cicatrical strictures in 47 patients is reduced. The method of operations performation and their advantage above one-brigade interventions is circumscribed. The complications, and also their matching with standard procedures of operative measures are reduced. The prospects for the development of two-brigade operations are construed. PMID- 15124475 TI - [Immediate and remote results of pylorus-preserving and pylorus-reducing operations in complex surgical treatment of perforated pyloric-duodenal ulcers]. AB - The results of surgical treatment of 224 patients with perforated pyloroduodenal ulcers were analyzed. 110 patients were operated using pyloruspreserving and pylorusreducing methods. The new method of pyloroduodenoplasty is suggested. Duodenogastral reflux after pyloruspreserving operations is arised in 4.5% of observations, after pylorusdestructive--in 46.7%. PMID- 15124476 TI - [Modern radiological method of diagnosis of respiratory foreign bodies and their complications in children]. PMID- 15124477 TI - [Historical aspects of thoracoscopic surgery]. PMID- 15124478 TI - [The role of interrelationship of neutrophils, endotheliocytes and macrophages in the etiology and course of septic-purulent state. Report 4. Intercellular interaction in inflammation]. PMID- 15124479 TI - [Fatal erosive hemorrhage due to tuberculous involvement of aorta]. PMID- 15124480 TI - [Iatrogenic intraperitoneal laceration of urinary bladder wall during trans urinary adenomectomy for benign hyperplasia of the prostate gland]. PMID- 15124481 TI - [Modification of gastric resection by Bihlroth I method]. PMID- 15124482 TI - [Method of catheterization of peripheral lymphatic vessel for prolonged application of preparations]. PMID- 15124483 TI - [Observation of segmental diverticulitis of small intestine with perforation, inter-loop abscess]. PMID- 15124484 TI - [Agenesis of gallbladder]. PMID- 15124485 TI - [Concurrent surgeries in children]. PMID- 15124486 TI - [Hyper-eosinophilia in the pleural exudate and peripheral blood in patient with exudative pleurisy]. PMID- 15124487 TI - [Sports and adolescence]. PMID- 15124488 TI - [Prostate cancer: view of the horizon]. PMID- 15124489 TI - [Technic of removing bladder clots]. PMID- 15124490 TI - [Sudden death: heart defibrillator as guardian angel]. PMID- 15124491 TI - [Decubitus ulcers: prevention is at the heart of the debate]. PMID- 15124492 TI - ["Let us defend the Institute of Rare Diseases" Interview by Helene Delmotte and Marie-France Guerel]. PMID- 15124493 TI - [Esther's teddy bear]. PMID- 15124494 TI - [Play helps you!]. PMID- 15124495 TI - [Somatropin]. PMID- 15124496 TI - [Biovigilance. (Decree no 2003-1206 of December 12, 2003]. PMID- 15124497 TI - Trouble in the air. PMID- 15124498 TI - Summer of the cicada. PMID- 15124499 TI - The time before dying. PMID- 15124500 TI - Improving cancer screening adherence: using the "teachable moment" as a delivery setting for educational interventions. PMID- 15124501 TI - Population-based hepatitis C surveillance and treatment in a national managed care organization. AB - OBJECTIVES: To use a national population-based automated claims database to study the testing rate, prevalence, and prescribing patterns for chronic hepatitis C. STUDY DESIGN: A retrospective descriptive study that analyzes medical and pharmacy automated claims from affiliated health plans in 4 regions of the United States. METHODS: Data were collected from 11 UnitedHealth Group-affiliated health plans (3.9 million members) from January 1, 1997, to December 31, 1999. Medical claims were used to identify persons tested for hepatitis C virus (HCV). Persons with chronic HCV were identified through medical and pharmacy claims. Patterns of drug use and treatment were analyzed, including prescribing physician specialty and proportion of patients receiving baseline and follow-up testing. RESULTS: Of 27,871 members tested for HCV (0.7%), 1869 (6.7%) were diagnosed as having chronic HCV. Tested patients were more likely to be female (odds ratio [OR], 1.1) and older (> or = 25 years; OR, 4.1). Of 3259 patients with HCV, most were male (OR, 1.8) and older (> or = 25 years; OR, 32.0). Of these patients, 33.6% (n = 670) of men and 25.2% (n = 319) of women received treatment. Combination therapy users were more likely to undergo baseline (OR, 4.8) and follow-up (OR, 6.2) testing compared with interferon alfa monotherapy users. CONCLUSIONS: Of the total population, 0.7% were tested for HCV, of whom 6.7% were diagnosed as having chronic HCV. Although women were more likely to undergo testing, prevalence and therapy rates for chronic HCV were higher in men. Most patients did not receive recommended baseline and follow-up testing, and the approximate 30% therapy rate suggested that many patients with HCV remain untreated. PMID- 15124502 TI - Comparing breast cancer case identification using HMO computerized diagnostic data and SEER data. AB - OBJECTIVE: To determine the sensitivity and positive predictive value (PPV) of computerized diagnostic data from health maintenance organizations (HMOs) in identifying incident breast cancer cases. STUDY DESIGN: An HMO without a cancer registry developed an algorithm identifying incident breast cancer cases using computerized diagnostic codes. Two other HMO sites with Surveillance, Epidemiology, and End Results (SEER) registries duplicated this case identification approach. Using the SEER registries as the criterion standard, we determined the sensitivity and PPV of the computerized data. METHODS: Data were collected from HMO computerized data-bases between January 1, 1996, and December 31, 1999. Surveillance, Epidemiology, and End Results data were also used. RESULTS: The overall sensitivity of the HMO databases was between 0.92 (95% confidence interval [CI], 0.91-0.96) and 0.99 (95% CI, 0.98-0.99). Sensitivity was high (range, 0.94-0.98), for the first 3 (of 4) years, dropping slightly (range, 0.81-0.94) in the last year. The overall PPV ranged from 0.34 (95% CI, 0.32-0.35) to 0.44 (95% CI, 0.42-0.46). Positive predictive value rose sharply (range, 0.18-0.20) after the first year to 0.83 and 0.92 in the last year because prevalent cases were excluded. Review of a random sample of 50 cases identified in the computerized data-bases but not by SEER data indicated that, while SEER usually identified the cases, the registry did not associate every case with the health plan. CONCLUSIONS: Health maintenance organization computerized databases were highly sensitive for identifying incident breast cancer cases, but PPV was low in the initial year because the systems did not differentiate between prevalent and incident cases. Health maintenance organizations depending solely on SEER data for cancer case identification will miss a small percentage of cases. PMID- 15124503 TI - Primary care practice and facility quality orientation: influence on breast and cervical cancer screening rates. AB - BACKGROUND: Despite the importance of early cancer detection, variation in screening rates among physicians is high. Insights into factors influencing variation can guide efforts to decrease variation and increase screening rates. OBJECTIVES: To explore the association of primary care practice features and a facility's quality orientation with breast and cervical cancer screening rates. STUDY DESIGN: Cross-sectional study of screening rates among 144 Department of Veterans Affairs (VA) medical centers and for a national sample of women. METHODS: We linked practice structure and quality improvement characteristics of individual VA medical centers from 2 national surveys (1 to primary care directors and 1 to a stratified random sample of employees) to breast and cervical cancer screening rates determined from a review of random medical records. We conducted bivariate analyses and multivariate logistic regression of primary care practice and facility features on cancer screening rates, above and below the median. RESULTS: While the national screening rates were high for breast (87%) and cervical cancer (90%), higher screening rates were more likely when primary care providers were consistently notified of specialty visits and when staff perceived a greater organizational commitment to quality and anticipated rewards and recognition for better performance. CONCLUSIONS: Organization and quality orientation of the primary care practice and its facility can enhance breast and cervical cancer screening rates. Internal recognition of quality performance and an overall commitment to quality improvement may foster improved prevention performance, with impact varying by clinical service. PMID- 15124504 TI - Health plan policies and programs for colorectal cancer screening: a national profile. AB - BACKGROUND: A consensus has emerged that average-risk adults 50 years of age or older should be screened for colorectal cancer (CRC). OBJECTIVES: To describe health plans' coverage policies, guidelines, and organized programs to promote CRC screening. STUDY DESIGN AND METHODS: Review of data from the National Cancer Institute Survey of Colorectal Cancer Screening Practices, administered to a national sample of health plans in 1999-2000. The survey inquired about coverage policies for fecal occult blood testing, sigmoidoscopy, colonoscopy, and double contrast barium enema; the nature of any guidelines the plan had issued to its providers on CRC screening; and systems for recruiting patients into screening and for tracking and reporting the results of screening and follow-up procedures. RESULTS: Of 346 eligible health plans, 180 (52%) responded. Nearly all health plans covered at least 1 CRC screening modality. Plans were most likely to cover fecal occult blood testing (97%) and least likely to cover colonoscopy (57%). Sixty-five percent had issued guidelines on CRC screening to providers. One quarter had a mechanism to remind patients that they are due for CRC screening, but fewer had systems for prompting providers, contacting noncompliant patients, or tracking completion of screening. CONCLUSIONS: Health plans have the ability to provide organizational infrastructure for a broad range of preventive services to well-defined populations. However, few health plans had all 3 essential CRC screening delivery components--coverage, guidelines, and tracking systems--in place in 1999-2000. PMID- 15124505 TI - Malpractice prevention, patient safety, and quality of care: a critical linkage. AB - There is growing evidence of a negative effect of the current American preoccupation with malpractice on efforts to reduce error, enhance safety, and improve other domains of quality. The use by some insurers of systems assessment and risk analysis programs, linked to rewards for performance--which, taken together, we term proactive risk management--offers an opportunity to enhance our focus on systems and to bring patient safety and malpractice risk reduction into close congruence with other quality improvement efforts. Given the increasing burden of malpractice, as well as the emerging concerns about patient safety, managed care organizations and their providers need to work together with malpractice insurers and quality improvement experts to refocus their efforts on creating systems improvement; driving measurement, analysis, and feedback; and developing incentives for performance that will align quality and risk management efforts and drive breakthroughs in quality, including patient safety. PMID- 15124506 TI - Verification of a decision analytic model assumption using real-world practice data: implications for the cost effectiveness of cyclo-oxygenase 2 inhibitors (COX-2s). PMID- 15124507 TI - Naming and taming our ethical dilemmas. PMID- 15124508 TI - Faculty matters. Patty J Hale. PMID- 15124509 TI - An ethics framework for nursing education on the Internet. AB - An ethics framework for the use of the Internet for nursing education, consultation, research, and practice is proposed. The proposed framework, discussed in terms of structure, process, and outcome, would guide policy for confidentiality, privacy issues, professionalism, ownership of intellectual property, accountability, and control of access. A goal would be to encourage creative problem solving without constant reference to authority. PMID- 15124510 TI - A development and implementation framework for Web-based nursing courses. AB - The demand for web-based nursing courses necessitates efficient, effective approaches to course development and implementation. The NursingWeb Framework addresses web-based nursing courses within the context of technological, legal/administrative, and curricular components and infrastructure and course specific structural features, processes/procedures, and outcomes. Aspects of the framework are illustrated using the web-based undergraduate nursing research course of the University of Maryland Baltimore's School of Nursing. PMID- 15124511 TI - Reflections on the role of faculty in distance learning and changing pedagogies. AB - The increasing number of nursing faculty teaching in distance education programs represents a paradigm shift that has implications for faculty role and changing pedagogies. This descriptive study investigated experiences of nursing faculty teaching web-based courses. Participants were drawn from eight nursing schools in the United States and Canada. Nineteen faculty discussed perceptions of teaching online in small-group teleconference interviews. Major categories identified were: faculty role issues, redesigning/rethinking courses, handling communications, developing partnerships, managing time, and dealing with technology. The core category was: redesigning pedagogies and rethinking faculty role for online teaching. Dimensional analysis was used to develop a matrix telling the story of the experience within the perspectives of antecedent conditions, context, strategies, and consequences. Results indicated that support systems, technology partnerships, and policies should be in place before redesign. The context of redesign was evident in moving from "on stage" to a virtual environment. Strategies used to redesign courses included collaboration, rethinking communications, and faculty development. Consequences had positive and negative outcomes with respect to their impact on faculty role, teaching approaches, and student/faculty relationships. PMID- 15124512 TI - Community-driven nursing transforming nursing curricula and instruction. AB - The aim of this study was to evaluate an innovative approach to teaching and learning community-based nursing. A hermeneutic research procedure was used to analyze data from individual interviews with undergraduate nursing students, faculty, and community preceptors. Two themes, or common meanings, of participation were identified: 1) Forming new partnerships: Experiencing community-driven nursing, and 2) Creating new visions: Thinking about community driven nursing. By describing and interpreting these themes, the authors show how nursing curricula and instruction should be transformed regardless of clinical setting. PMID- 15124513 TI - The student/faculty international exchange: responding to the challenge of developing a global perspective in nursing education. AB - A student/faculty international exchange is an effective means for nursing programs to respond to the challenge of developing a global perspective in nursing education. This article explores the use of a student/faculty exchange as an intervention to help facilitate an international partnership. Bandura's social learning theory provides the conceptual framework to model the exchange, a collaborative effort that offers faculty and students the opportunity to acquire an international view of health care by comparing and sharing nursing data across populations, settings, and geographic areas. Strategies for developing a successful exchange program, obstacles involved in the process, and evaluation methods are addressed. PMID- 15124514 TI - Hallmarks of excellence in nursing education. PMID- 15124515 TI - The new pill pushers. PMID- 15124516 TI - The roots of war. PMID- 15124517 TI - Driven to distraction. PMID- 15124518 TI - The temperamental mind. PMID- 15124519 TI - Preventing fires, igniting questions. PMID- 15124520 TI - Perspectives. Determined drug-card players face barriers on take-up. PMID- 15124521 TI - The first Nobel Peace Prize, Henry Dunant (founder of the International Red Cross) and his "Memoirs". AB - To celebrate the memory and work of Henry Dunant, on the centenary of the presentation of the first Nobel Peace Prize, rightly awarded to Dunant for his having founded the institution of the International Red Cross, this paper presents the reader with some insights into his activities and sufferings, his trials and tribulations, and the hope and strength of his character. The ceaseless efforts made by Dunant to bring about the Institution which today represents Hope for so many suffering people who are silent victims of wars and atrocities, are fleetingly presented. The authors' intention is to give due recognition to Dunant for his work, and to highlight the humanity and the moral and social worth of the face behind the International Red Cross. PMID- 15124522 TI - Embryonic discourse: abortion, stem cells and cloning. AB - This Article interprets the debate about abortion and the debate about embryonic research and therapeutic cloning as aspects of a larger history of ideas. The Article suggests that embryos increasingly stand for different truths in discourse about abortion on the one-hand and about embryonic stem cell research and therapeutic cloning on the other. More specifically, the Article suggests that the contemporary debate about the meaning of the embryo in the context both of abortion and of embryonic research bespeaks a widespread transformation in Western, and especially American, society during the last three or four decades. At base, that transformation involves displacement of an understanding of personhood, particularly in domestic settings that depended on the submersion of individualism with an understanding of personhood that values autonomous individuality and that envisions community as the consequence of individuals' distinct choices rather than as a pre-existing, hierarchically structured whole. PMID- 15124523 TI - Adult stem cells. PMID- 15124524 TI - Production of hypoxia-induced corneal edema in aged eyes. AB - PURPOSE: Corneal thickness assessment is a common clinical procedure applied in corneal and contact lens care. This study aims to investigate the effect of age on hypoxia-induced corneal swelling. METHODS: Eighteen male subjects were equally divided into the younger [(23.7 +/- 0.8) years old] and older [(74.4 +/- 2.5) years old] groups. Each subject wore a thick soft contact lens (uniform thickness of 0.3 mm) on the left cornea. With the contact lens in place, the baseline central corneal thickness was measured using a specially designed photo pachometer. The lens was then patched behind the closed eyelids, producing an extremely hypoxic stress to the cornea. The change in central corneal thickness was monitored every 20 minutes with momentary disruptions to the hypoxic stress over the next 2 hours. The increase in thickness was taken as an index of corneal edema. The rate of change in corneal thickness, as derived from a non-linear mathematical model, was compared between groups. RESULTS: The corneal thickness of both age groups increased significantly with time (P < 0.0001). The mean corneal swelling constant for the older subjects was 16.5 x 10(-3) (S.E.M. = 2.65 x 10(-3)) and the value for the younger subjects was 46.5 x 10(-3) (S.E.M. = 3.25 x 10(-3)). The difference was statistically significant (P < 0.0001). CONCLUSION: Aging cornea has a slower hypoxia-induced edema response compared with the younger group. Whether it is caused by a decreased corneal lactate production or an increased resistance to physical expansion deserves further investigation. PMID- 15124525 TI - Comparison of the inflammatory reaction to laser photolysis and conventional phacoemulsification. AB - PURPOSE: To compare the postoperative inflammatory reaction induced by laser photolysis and conventional phacoemulification. METHODS: Anterior chamber flare and cells were measured preoperatively and 1 day, 1 week, 1 month and 3 months postoperatively in two groups. In group 1, thirty-two eyes underwent laser photolysis. In group 2, thirty-five eyes underwent conventional phacoemulification. RESULTS: Anterior chamber flare and cell measurements had no significant difference between two groups preoperatively and 1 day, 1 week, 1 month and 3 months postoperatively. Flare and cell values returned to preoperative values by the third month. CONCLUSIONS: Postoperative inflammatory response to laser photolysis was equal to conventional phacoemulsification, and recovery occurred in 3 months postoperatively. PMID- 15124526 TI - Clinical analysis of 106 cases with elevated intraocular pressure in thyroid associated ophthalmopathy. AB - PURPOSE: To summarize the clinical manifestation of thyroid-associated ophthalmopathy (TAO) with elevated intraocular pressure (IOP), and to analyze the contributing factors. METHODS: One hundred and six cases (188 eyes) of ocular hypertension in 339 cases (597 eyes) with TAO were collected from 1994 to 2001 and their clinic manifestations were summarized and analyzed. RESULTS: It was demonstrated that the incidence of ocular hypertension in TAO was 31.3%, and was more frequently found in the male than in the female. The elevated IOP in TAO was found to be partially related to compression of the eyeball by enlarged extraocular muscles, the elevated intraorbital pressure as result of the proliferation of intraorbital connective tissue and the enlargement as well as swelling of extraocular muscles. It was also related to the severity of TAO other than the course of TAO. The ocular hypertension in most cases can be controlled with reduction of IOP by methylprednisolone or orbital decompression. CONCLUSION: Specific clinical features were found in TAO patients with ocular hypertension. It should be differentiated with primary glaucoma. The IOP in most cases can be controlled by prompt and effective treatment of TAO. PMID- 15124527 TI - Clinical features of choroidal metastases from carcinoid tumour. AB - PURPOSE: To report ophthalmologic and angiographic features of choroidal metastases from carcinoid tumor and analyze their common clinical manifestation. METHODS: Ophthalmologic examinations and fundus fluorescein angiography (FFA) were performed in 30 patients suffered from carcinoid tumor, and four patients diagnosed of breast cancer (2 cases), lung cancer (1 case) and maxillary sinus cancer (1 case) were confirmed with choroidal metastases. RESULTS: Choroidal metastases were found as the initial manifestations of the malignant tumors on 2 patients whose initial and chief complaints were decreasing vision, their fundus lesions were mainly presented in the posterior pole and FFA showed high density of fluorescence of the lesions. CONCLUSIONS: This study indicated choroidal metastasis might be the first sign of metastases for patients with cancer. For patients with unknown metastasic cancers, examinations of the choroids may be useful for diagnosis and prognosis. PMID- 15124528 TI - Angioleiomyoma of the ciliary body: a case report. AB - PURPOSE: To report a rare case of angioleiomyoma of the ciliary body METHODS: The clinical manifestation, imaging findings, histopathologic characteristics were analyzed in a 32-year-old male patient with angioleiomyoma of the ciliary body. RESULTS: The tumor was removed intact with local resection. Histopathologic examination revealed that the tumor was full of vessels and it was composed of spindle cells with abundant cytoplasm. Immunohistochemical studies showed positive for SMA and Desmin and negative for S100 and HMB-45. CONCLUSIONS: Angioleiomyoma of the ciliary body is a rare tumor that can be successfully treated with local surgical resection in this area. It needs to be differentiated from other tumors, especially malignant melanoma. PMID- 15124529 TI - A case of choroidal tubercles. AB - PURPOSE: To report a case of choroidal tubercles in a miliary tuberculosis boy. METHOD: Clinical features description. RESULTS: A 14-year-old boy was found to have multifocal choroidal tubercles. Angiography was performed. He was followed up for 12 months. The choroidal lesions regressed after using anti-tuberculosis drugs for 8 months. Pigment changes remained. CONCLUSION: Tuberculosis may present as a posterior segment inflammation. In miliary tuberculosis, choroid is also a target tissue. PMID- 15124530 TI - Characteristics of optic nerve damage induced by chronic intraocular hypertension in rat. AB - PURPOSE: To set up the Sharma's chronic intraocular hypertension model and investigate the intraocular pressure (IOP) as well as the optic nerve damage of this model in rat. METHODS: The operations of the chronic intraocular hypertension model were performed as described by Sharma in 60 Male Lewis albino rats. IOP was measured using the Tono-Pen XL immediately after surgery and then at 5 day, 2 week or 4 week intervals. Cresyl violet staining of whole-mounted retinas was used to label retinal ganglion cells (RGCs), then RGCs were counted. Paraphenylenediamine (PPD) staining was performed in the semi-thin cross sections of optic nerve of rat, in order to know whether the axons of optic nerve were degenerated or not. RESULTS: There were 47 rats with higher IOP after the episcleral veins cauterized in 60 rats. The ratio of elevated IOP was 78.3%. The IOPs were stable in 4 weeks. After cresyl violet staining, the RGCs loss was 11.0% and 11.3% was found in the central and peripheral retina respectively after 2 weeks of increased IOP. After 4 weeks of increased IOP, the loss of RGCs was 17% for the central retina and 24.6% for the peripheral retina. In the retinas without higher IOP, there was no loss of RGCs. PPD staining showed that optic nerve of rat with about 5.3% damage of axons located at the superior temporal region. Region of affected optic nerve 1 mm posterior to the globe by light microscope showed evidence of damaged axons with axonal swelling and myelin debris. CONCLUSION: Sharma's chronic intraocular hypertension model is a reproducible and effective glaucoma model, which mimics human glaucoma with chronically elevation IOP and induced RGCs loss and damage of optic nerve. PMID- 15124531 TI - Induction of heat shock protein 72 in RGCs of rat acute glaucoma model after heat stress or zinc administration. AB - PURPOSE: To investigate the dynamics of heat shock protein 72 (HSP72) expression in retinal ganglion cells (RGCs) in rat model of acute glaucoma treated with heat stress or intraperitoneal injection of zinc sulfate. METHODS: Twenty-seven male Wistar rats were used to make acute glaucoma models. Five others served as normal control. Acute glaucoma models were made by intracameral irrigation in the right eyes with balanced salt saline (BSS) at 102 mmHg for 2 hours. Nine model rats were killed at different intervals after intracameral irrigation without treatment, which served as damage control. Ten were treated with heat stress 40 degrees C-42 degrees C, and 8 were used for zinc sulfate administration 2 days posterior to intracameral irrigation. Treated model rats were sacrificed at designed intervals after treatment. Right eyes were enucleated immediately, and the retinas were dissected for Western blot. RESULTS: No HSP72 was found in RGCs of normal Wistar rats. In damage control group, slight HSP72 was detected during 6-36 hours posterior to intracameral irrigation. HSP72 was detected significantly expressed in RGCs of both heat shock group and zinc sulfate group. But the dynamics of HSP72 production were quite different in these two treated groups. In heat shock group, HSP72 appeared at the sixth hour after treatment, and increased gradually until its peak production emerged at the 48th hour. HSP72 vanished 8 days later after treatment. In zinc sulfate group, HSP72 expression began 24 hours later after zinc administration, and reached its highest level at the 72th hour posterior to treatment. HSP72 expression then decreased slowly, and disappeared 21 days later after treatment. CONCLUSION: HSP72 can be induced in RGCs of rat acute glaucoma models with heat stress or zinc sulfate administration. But the dynamics of the HSP72 induction in those two groups were quite different. PMID- 15124532 TI - Experiment study of retinal ultrastructure after intravitreal FK506. AB - PURPOSE: To investigate the retinal toxicity of FK506 by intravitreal administration. METHODS: Twenty-two eyes of 14 New Zealand rabbits were investigated. FK506 at concentrations of 5,25 and 50 micrograms/eye was injected into the vitreous cavities respectively. The control eyes were received mixed solution of balanced salt and ethanol. All eyes were examined by tonometry, slit lamp and indirect ophthalmoscopy preoperatively and postoperatively at the 1st, 3rd, 7th, and 14th day respectively. In the final examination, all eyes were enucleated and processed for light and electron microscopy. RESULTS: No evidence of toxic reaction was seen in the eyes received 25 micrograms FK506 or less of FK506. Several eyes received 50 micrograms FK506 and control eyes developed conjunctival congestion and slightly bloody exudates in anterior chamber which may be related to irritation of ethanol. Two of five eyes received 50 micrograms developed transient vitreous opacities. Electron microscopically, the mitochondria of the photoreceptor cells were swelled in the eyes treated with 50 micrograms FK 506. CONCLUSION: It is safety with intravitreal FK506. There are no irritation and toxicity to the rabbits eyes with the intravitreal doses of 25 micrograms FK506 or less. The doses of 50 micrograms FK506 are proved to be toxic to the retina. PMID- 15124533 TI - Influence of IL-1 beta and TNF-alpha on Fas expression of human retinal pigment epithelial cells in vitro. AB - PURPOSE: To observe Fas expression change of cultured human retinal pigment epithelial (RPE) cells by IL-1 beta and TNF-alpha. METHODS: With flow cytometry, immunohischemistry, and color imaging system, Fas expressions by exposure to IL-1 beta and/or TNF-alpha were measured. RESULTS: The gray degree values of Fas expression were 67.5 +/- 6.1 in IL-1 beta + TNF-alpha-treated group, 80.1 +/- 9.2 in IL-1 beta-treated group, and 70.4 +/- 6.4 in TNF-alpha-treated group, respectively. There were significant differences (P < 0.005) compared with control group (107.0 +/- 10.2). Flow cytometry showed that 15.0% cultured human RPE cells expressed Fas. Fas-positive in IL-1 beta, TNF-alpha, and IL-1 beta + TNF-alpha-treated groups expressed was 28.1%, 34.5%, and 65.2%, respectively. CONCLUSION: IL-1 beta, TNF-alpha, and combining both of them can up-regulate Fas protein expression, which may contribute to more Fas (+) cells in proliferative vitreoretinopathy (PVR). Minimizing this process by means of inducing apoptosis of Fas (+) proliferative cells of Fas/FasL pathway is a future preventive and therapeutic possibility for PVR. PMID- 15124534 TI - Muscarinic receptor agonists protect cultured bovine trabecular meshwork cells against apoptosis induced by dexamethasone. AB - PURPOSE: To study whether muscarinic receptor agonists can protect cultured bovine trabecular meshwork cells against apoptosis induced by dexamethasone. METHODS: The third to fifth passages of bovine trabecular meshwork cells were grown to confluence and incubated for 1-14 days in growth media with dexamethasone or pretreatment of pilocarpine or carbachol. The cultures were evaluated for apoptosis by phase-contrast microscopy, fluorescence microscopy, DNA laddering and flow cytometric analysis. RESULTS: Dexamethasone (0.24-0.96 mmol.L-1) induced apoptosis of trabecular meshwork cells in a dose and time dependent manner. Before 0.48 mmol.L-1 dexamethasone-treatment, 1.84 mmol.L-1 of pilocarpine or 2.74 mmol.L-1 of carbachol added could significantly reduce apoptotic percentage. CONCLUSION: Muscarinic receptor agonists can protect cultured bovine trabecular meshwork cells against apoptosis induced by dexamethasone. PMID- 15124535 TI - Quantitative measurements of the decentration distance of four kinds of intraocular lens. AB - PURPOSE: To investigate a quantitative method to measure decentration distance(DD) and analyze the DD of four kinds of intraocular lenses (IOLs). METHODS: The pseudophakic eye photos were taken under the slit lamp after the IOLs were implanted a year or longer. Then the images were inputted into a computer and the Photoshop software was applied to analyze the DD. Four kinds of IOLs were analyzed, including AcrySof MA60BM (11 eyes), Allergan SA40N (11 eyes), AcrySof SA60AT/SA30AL (5 eyes), Corneal Quattro (4 eyes). RESULTS: The mean value of DD was (0.2643 +/- 0.1574) mm in the AcrySof MA60BM group, (0.3536 +/- 0.1719) mm in the Array SA40N group, (0.3099 +/- 0.1529) mm in the SA60AT/SA30AL group, and (0.3261 +/- 0.1877) mm in the Quattro group. The difference of the DD values among these 4 kinds of IOLs had no statistic significance (F = 0.506, P = 0.681) and the values were not equal to zero (t = 10.508, P < 0.001). CONCLUSION: The Photoshop method is a simple and efficient way for DD analysis. The modern foldable posterior-chamber-IOLs have good centration performances when implanted into the capsular bags. PMID- 15124536 TI - Expression of CD44 in cultured human trabecular meshwork cells. AB - PURPOSE: To determine whether cultured human trabecular meshwork cells express CD44 and to discuss their possible relationship with primary open angle glaucoma. METHODS: Human trabecular meshwork cells were cultured in DMEM/F12 media. Total RNAs from the cells were extracted with Trizol reagent. Messenger RNA expression of CD44 in human trabecular meshwork cells was examined by using reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Expression of CD44 was confirmed by Western-blotting and immunofluorescent microscopy. Effect of CD44 specific antisense oligonucleotide on adhesion of trabecular meshwork cells to hyaluronate was determined by MTT assay. RESULTS: A single RT-PCR product whose size was 471 bp was obtained. A band about 80kD was stained by Western-blot. Immunofluorescent examination of expression of CD44 on the cell surface was positive and reactions were mainly localized in cell membranes. Adhesion of trabecular meshwork cells to hyaluronate was inhibited by CD44-specific antisense oligonucleotide. CONCLUSIONS: Cultured human trabecular meshwork cells express CD44. CD44 may play a role in pathogenesis of primary open angle glaucoma. PMID- 15124537 TI - Fundus analysis and visual prognosis of macular hemorrhage in pathological myopia without choroidal neovasculopathy. AB - PURPOSE: To analysis and evaluate the fundus characteristics and visual prognosis of macular hemorrhage in pathological myopia without choroidal neovasculopathy. METHODS: Thirty-seven patients (38 eyes) of pathological myopia with macular hemorrhage and without choroidal neovascularization (CNV) underwent color photograph and fundus fluorescein angiography (FFA) examinations. Indocyanine green angiography (ICGA) was also performed on 11 patients (11 eyes). Follow-up ranged from 3 to 21 months. RESULTS: The macular hemorrhage in pathological myopia without CNV demonstrated oval, less than 1PD, without edema and exudation. Lacquer cracks appeared at the site of previous subretinal bleeding in 84.2% of the eyes. The visual acuities were improved in 81.6% of eyes during the follow-up period. ICGA revealed linear hypofluorescence in 7 of 11 eyes (63.6%), indicating a ruptured Bruch's membrance at the onset of subretinal bleeding. CONCLUSION: A rupture of choriocapillaris complex and Bruch's membrane causes macular hemorrhage of pathological myopia without CNV, leading to the formation of a new lacquer crack. Its prognosis is favorable. PMID- 15124538 TI - [Endometrial carcinoma: which radiotherapy for which patient?]. AB - The endometrial carcinoma is the third most common cancer in women. The endometrioid adenocarcinomas represent 75% to 80% of the pathologic sub-types, and 90% of the lesions are stages I or II at diagnosis. The treatment strategies are based upon surgery and radiotherapy, but the respective place of external irradiation and vaginal cuff brachytherapy is not strictly established. The role of the pelvic lymphadenectomy and the role of chemotherapy for aggressive histologies or locally advanced diseases remain also controversial issues. The question "which radiotherapy for which patient?" is still a current question, however, the answers we will bring do not permit yet to remove all uncertainties. PMID- 15124539 TI - [Dose-response relationship in radiotherapy: an evidence?]. AB - The dose-response relationship is a fundamental basis of radiobiology. Despite many clinical datas, difficulties remain to demonstrate a relation between dose and local control: relative role of treatment associated with radiation therapy (surgery, chemotherapy, hormonal therapy), tumor heterogeneity, few prospective randomized studies, uncertainty of local control assessment. Three different situations are discussed: tumors with high local control probabilities for which dose effect is demonstrated by randomized studies (breast cancer) or sound retrospective datas (soft tissues sarcomas), tumors with intermediate local control probabilities for which dose effect seems to be important according to retrospective studies and ongoing or published phase III trials (prostate cancer), tumors with low local control probabilities for which dose effect appears to be modest beyond standard doses, and inferior to the benefit of concurrent chemotherapy (lung and oesophageal cancer). For head and neck tumors, the dose-response relationship has been explored through hyperfractionation and accelerated radiation therapy and a dose effect has been demonstrated but must be compared to the benefit of concurrent chemotherapy. Last but not least, the development of conformal radiotherapy allow the exploration of the dose response relationship for tumors such as hepatocellular carcinomas traditionally excluded from the field of conventional radiation therapy. In conclusion, the dose response relationship remains a sound basis of radiation therapy for many tumors and is a parameter to take into account for further randomized studies. PMID- 15124540 TI - [Respiration-gated radiotherapy: current techniques and potential benefits]. AB - Respiration-gated radiotherapy offers a significant potential for improvement in the irradiation of tumor sites affected by respiratory motion such as lung, breast and liver tumors. An increased conformality of irradiation fields leading to decreased complications rates of organs at risk (lung, heart...) is expected. Respiratory gating is in line with the need for improved precision required by radiotherapy techniques such as 3D conformal radiotherapy or intensity modulated radiotherapy. Reduction of respiratory motion can be achieved by using either breath hold techniques or respiration synchronized gating techniques. Breathhold techniques can be achieved with active, in which airflow of the patient is temporarily blocked by a valve, or passive techniques, in which the patient voluntarily breath-hold. Synchronized gating techniques use external devices to predict the phase of the respiration cycle while the patient breaths freely. These techniques presently investigated in several medical centers worldwide. Although promising, the first results obtained in lung and liver cancer patients require confirmation. Physical, technical and physiological questions still remain to be answered. This paper describes the most frequently used gated techniques and the main published clinical reports on the use of respiration gated radiotherapy in order to evaluate the impact of these techniques. PMID- 15124541 TI - [Surgical robotics, short state of the art and prospects]. AB - State-of-the-art robotized systems developed for surgery are either remotely controlled manipulators that duplicate gestures made by the surgeon (endoscopic surgery applications), or automated robots that execute trajectories defined relatively to pre-operative medical imaging (neurosurgery and orthopaedic surgery). This generation of systems primarily applies existing robotics technologies (the remote handling systems and the so-called "industrial robots") to current surgical practices. It has contributed to validate the huge potential of surgical robotics, but it suffers from several drawbacks, mainly high costs, excessive dimensions and some lack of user-friendliness. Nevertheless, technological progress let us anticipate the appearance in the near future of miniaturised surgical robots able to assist the gesture of the surgeon and to enhance his perception of the operation at hand. Due to many in-the-body articulated links, these systems will have the capability to perform complex minimally invasive gestures without obstructing the operating theatre. They will also combine the facility of manual piloting with the accuracy and increased safety of computer control, guiding the gestures of the human without offending to his freedom of action. Lastly, they will allow the surgeon to feel the mechanical properties of the tissues he is operating through a genuine "remote palpation" function. Most probably, such technological evolutions will lead the way to redesigned surgical procedures taking place inside new operating rooms featuring a better integration of all equipments and favouring cooperative work from multidisciplinary and sometimes geographically distributed medical staff. PMID- 15124542 TI - [Automatization and robotics of the set-up and treatment of patients irradiated for brain and base ot the skull tumors]. AB - Progresses of the three-dimensional imageries and of the software of planning systems makes that the radiotherapy of the tumours of brain and the base of skull is increasingly precise. The set-up of the patients and the positioning of the beams are key acts whose realization can become extremely tiresome if the requirement of precision increases. This precision very often rests still on the visual comparison of digital images. In the near future, the development of the automated systems controlled by robots should allow a noticeable improvement of the precision, safety and speed of the patient set-up. PMID- 15124543 TI - [Intensity modulated radiotherapy (IMRT) in France: The boost of the national funding for the new expensive innovative technologies (STIC 2001 and 2002)]. AB - The STIC 2001 and STIC 2002 projects intend to allow the implementation and the assessment of Intensity Modulated Radiation Therapy in France. IMRT is an innovative technique in which the high-dose radiation volume conforms to an accurately defined target volume with less morbidity to the surrounding normal tissues. The main medical objectives of the projects are (1) to improve the therapeutic index while decreasing acute toxicity and late sequelae (mainly xerostomia and acute mucite for head and neck tumors), which allows an increase in the radiation dose to the tumor and then a better tumor control; (2) to propose a salvage treatment to patients who locally recurred in previously irradiated sites; (3) to determine the optimal treatment guidelines for a safe use of the technique in clinical routine. Our projects also aim at comparing IMRT and 3D conformal treatments on the one hand (STIC 2001), and IMRT and conventional treatments on the other hand (STIC 2002), with regard to costs. As a matter of fact, the use of IMRT is presently limited in France because its implementation requires high investment and personnel costs. The seventeen French Regional Cancer Centres involved in the two projects intend to study the additional cost of the use IMRT in comparison with the use of standard techniques, which appears to be a step for a wide use of this technique in France. Each of the studies is two-year prospective, and includes patients with head and neck tumors treated with a curative intend (post operative or exclusive treatments for STIC 2002 and STIC 2002), and patients with a prostate cancer (STIC 2001). PMID- 15124544 TI - [Radiation protectants of the crystalline lens]. AB - During more than a half of century, numerous compounds have been tested in different models against radiation-induced cataract. In this report, we will review the radioprotectors that have been already tested for non-human crystalline lens protection. We will focus on the most important published studies in this topic and the mechanisms of cytoprotection reported in vitro and in vivo from animals. The most frequent mechanisms incriminated in the cytoprotective effect are: free radical scavenging, limitation of lipid peroxidation, modulation of cycle progression increase of intracellular reduced glutathion pool, reduction of DNA strand breaks and limitation of apoptotic cell death. Amifostine (or Ethyol) and anethole dithiolethione (or Sulfarlem), already used clinically as chemo- and radioprotectants, could be further tested for ocular radioprotection particularly for radiation-induced cataract. PMID- 15124545 TI - [New developments in oncologic hyperthermia]. AB - Hyperthermia between 40 and 45 degrees C is now recognized to radiosensitive tumors locally increasing local control. The more recent technological progress show the possibility of heating at depth while controlling temperature by opened NMR. In Germany and Netherland, hyperthermia is a standard treatment for cervix carcinomas and for some sarcomas. Many on going clinical trials has been recently opened and a European network of excellence "Eurotherm" is proposed. More recently thermo-sensitive liposomes has been proposed in the US for carrying and targeting not only chemotherapeutics agents but also gene therapies. In France, these deep heating techniques are not applied. Today, only microwave superficial hyperthermia, thermo ablation using either radiofrequencies or high focussed ultrasound, and loco regional heating by intraperitoneal perfusion or by external circulation for tumors of the extremities, are in clinical evaluation. Is appears suitable to define clinical French teams, that could evaluate the clinical use of deep heating by the more recent developed techniques. PMID- 15124546 TI - [Cancer of the testis: role of radiotherapy in 2003]. AB - Germ-cell tumors of the testis are rare tumors of the young adult. Half of them are seminoma. The majority of patients have disease limited to the testis. Radiotherapy still remains the standard treatment of these patients. Almost all patients are cured by orchidectomy and radiotherapy on the lomboaortic area extended to homolateral iliac area. The dose is 24 to 30 Gy in a standard fractionation. Different studies are ongoing to reduce the irradiation field (omission of the pelvic irradiation), to decrease irradiation dose (to 20 Gy). Other treatment options are strict surveillance and adjuvant carboplatin based chemotherapy. None of these options are standard treatments. A strict attention must be directed on controlateral germ-cell tumors and second cancers. PMID- 15124547 TI - [Radiotherapy of testicular seminoma: changes over the past 10 years]. AB - Radiotherapy is generally considered as the standard treatment for most testicular seminomas. However, there have been substantial changes in the management of these tumours over the past few years. In early seminoma, there is a trend towards a decrease in treatment intensity or even towards therapeutic abstention (i.e. surveillance); whereas in advanced cases, combination chemotherapy is taking over from radiotherapy. In stage I, where cure rates are almost 100%, the limiting of the lymph node area to be irradiated and decrease of the dose to 20-25 Gy was followed by very low long-term toxicity rates, and a very small risk of infertility, without compromising the overall prognosis. Surveillance is an acceptable alternative to postoperative radiotherapy. However, the risk of nodal relapse is around 18-20%. With surveillance, the frequency and duration of follow-up is increased in comparison to the same with postoperative radiotherapy, with higher cost. In stage IIa, radiotherapy remains the standard but recent studies have shown that limiting the nodal volume to the paraortic area is justified as in stage I. In stage IIb and higher, combination chemotherapy is almost always given. However, the association between carboplatin and radiotherapy represents an efficient and well-tolerated alternative. Late tissue damage and the risk of decrease in fertility are minimized with novel radiotherapeutic approaches. However, the occurrence of second cancers in the long term is a matter of concern. It is possible though, that patients with seminoma have a tendency per se to develop second cancers. The prognosis of cryptorchid seminoma and of HCG-producing seminoma has been the subject of controversy but recent large studies have demonstrated that stage for stage, the cure rates are similar to those of other seminomas. PMID- 15124548 TI - [Verification of the dose delivered to the patient by means of TLD, SC, PID. What future?]. AB - Among the different possibilities to check the accuracy of the treatment delivered, only in vivo dosimetry ensures the precision of the dose delivered to the patient during the treatment. In 1970-1980, Ruden assessed the use of thermoluminescent dosimetry to perform in vivo measurements at Radiumemmet in Stockholm. Straightforward in its principle but demanding in its implementation, thermoluminescent dosimetry has largely been used. Today, thanks to the work of Rikner, the use of semiconductor detectors allows the general implementation of in vivo dosimetry. Tomorrow, we will use electronic portal imaging device to verify the geometrical patient setup and the dose delivery at the same time. Its implementation remains complex and will need the development of algorithms to compute exit dose or midplane dose using portal in vivo dosimetry. First clinical results show that portal imaging is an accurate alternative for conventional in vivo dosimetry using diodes. PMID- 15124549 TI - [Lymphatic mapping and inguinal sentinel lymph node biopsy in anal canal cancers to avoid prophylactic inguinal irradiation]. AB - Thirty-five patients with clinically N0 cancers of the canal anal, 33 epidermoid carcinomas et 2 melanomas were histologically staged with inguinal sentinel lymph node biopsy (ISN). With the combined technique, blue dye and radiocolloid the ISN was identified in 100% of the cases. The ISN was invaded in 7 cases/33 for epidermoid tumors and 2/2 for melanomas. After 18 months of follow-up, no inguinal recurrence could be seen in ISN pN0 cases. In conclusion, ISN biopsy is a reliable procedure to stage anal canal cancers. It should prevent unnecessary prophylactic inguinal irradiation for pN0 ISN. Inguinal irradiation is only indicated in pN1 ISN. PMID- 15124550 TI - [Radiotherapy of carcinomas of the anal canal. Tenon Hospital experience]. AB - Since 1980, curative-intent radiation therapy of epidermoid carcinoma of the anal canal is the standard first line treatment. The combined concomitant chemotherapy and radiation therapy is presently established for locally advanced tumors more than 4 cm in length and/or with nodal involvement. We report the Tenon hospital experience since 1972 concerning the long term results after radiation therapy, the modifications of the radiation technique, and the evolution of treatment strategy. PMID- 15124551 TI - [Rationale and progress of the phase III trial: intensification of the treatment of locally advanced squamous cell carcinoma of the anal canal]. AB - Concomitant radiotherapy (5FU-MMC) was proved to be useful in locally advanced anal canal carcinoma. Nevertheless, it remains 30% of failures after this conservative treatment. The tolerance and efficiency of a neoadjuvant chemotherapy (5-FU-CDDP) were validated by a phase II trial including 80 patients, which obtained 73% of colostomy free survival and 70% of relapse free survival at 3-year follow-up. Its usefulness is studied in an ongoing phase III trial, as well as the dose escalation of the boost, from 15 Gy to 25-25 Gy. The results of the 101 first included patients are studied by an intermediate analyze. In July 2003, 222 patients were enrolled by 33 investigating centres out of the 350 planned patients until the end of the trial in December 2004. PMID- 15124552 TI - [MRI of the breast. Techniques and current values]. AB - Breast MR imaging and PET allow physiopathological tissular approach of breast tumors through tumoral angiogenesis. All these series published agreed in the high sensitivity of breast MRI for detecting invasive breast cancers whereas the sensitivity varies from 60 to 85% in detecting ductal carcinoma in situ. However, its specificity reported varies between 60 and 97%, mainly due to some entities such as fibroadenomas and fibrocystic disease. Clinical indication are the diagnosis of local relapses, the evaluation of pre-operative chemotherapy, the diagnosis of an isolated axillary lymphadenopathy, pre-operative evaluation of breast cancer and follow up of breast prothesis. PMID- 15124553 TI - [Histological margin and residual disease assessment for breast carcinoma]. AB - Margin and histological size of ductal in situ carcinoma or intraductal component of an infiltrative carcinoma are important prognostic factors to predict presence/absence as well as amount of residual tumor burden. Their evaluation requires standardized pathological analysis. These factors should be interpreted in clinical and radiological context. PMID- 15124554 TI - [Accelerated partial breast irradiation following breast conservative surgery. A review of the literature]. AB - After breast-conservative surgery, radiation therapy delivered to the whole breast to 50 Gy with an additional boost of 10 to 16 Gy is the standard of care. Based upon data showing that the vast majority of the recurrences occur within and surrounding the original tumor site and in order to reduce the morbidity of whole breast radiotherapy and to give the treatment in a shorter time, partial breast irradiation has been developed by several institutions. Partial breast radiotherapy is given by intraoperative radiotherapy (photons or electrons), or brachytherapy (low dose rate or high dose rate) or external radiotherapy using 3D conformal radiation therapy. This is a review of the literature data on this topic. PMID- 15124556 TI - [French Society of Oncologic Radiotherapy. Membership list]. PMID- 15124555 TI - [Partial breast irradiation: high dose rate peroperative brachytherapy technique using the MammoSite]. AB - In the conservative management of breast cancer, radiation therapy delivering 45 to 50 Gy to the whole breast, in 4.5 to 5 weeks, followed by a booster dose of 10 to 20 Gy is the standard of care. Based on the numerous studies which have reported that the local recurrences occurs within and surrounding the primary tumor site and in order to decrease the treatment duration and its morbidity, partial breast irradiation using several techniques has been developed. Partial irradiation may be considered as an alternative local adjuvant treatment for selected patients with favorable prognostic factors. Using external beam radiation therapy, the 3D-conformal technique is appropriate to deliver the whole dose to a limited volume. In UK, an intraoperative technique using a miniature beam of low energy of x-ray (50 Kv) has been developed (Targit). Milan's team have developed an intraoperative electrons beam radiotherapy using a dedicated linear accelerator in the operative room. In USA and Canada the MammoSite has been advised for clinical use in per-operative brachytherapy of the breast. These two last techniques are currently compared in phase III randomised studies to the standard whole breast irradiation followed by a tumour bed booster dose. In this review we will focus on the MammoSite technique and will describe the per operative implantation procedure, radiological controls ad dosimetric aspects. PMID- 15124557 TI - Nobel winner pushes for women's rights. PMID- 15124558 TI - Kaiser study shows media rarely focus on most affected by HIV. PMID- 15124559 TI - Shareholders applaud Coke for Africa HIV/AIDS initiative. PMID- 15124560 TI - Religious groups stand firm for action on HIV/AIDS in Africa. PMID- 15124561 TI - Prisons. Fear of HIV exposure insufficient to uphold indifference claim. PMID- 15124562 TI - EMT not required to prove HIV exposure for disability claim. PMID- 15124563 TI - Elderly and AIDS. CDC reports jump in number of older Americans with AIDS. PMID- 15124564 TI - Felony exposure. Man jailed for failing to disclose HIV-positive status. PMID- 15124565 TI - Prisons. Inmate's claim he was refused AIDS drugs survives challenge. PMID- 15124566 TI - HIV cases take time, resources to make ADA claim. PMID- 15124567 TI - ADA. How to define major life activities in HIV disability cases. PMID- 15124568 TI - ADA. 5 ways to explore HIV's effect on limiting life activity. PMID- 15124569 TI - Negligence. Government cleared in disabled man's AIDS-related death. PMID- 15124570 TI - Confidentiality. Inmates given access to medical records in disclosure dispute. PMID- 15124571 TI - Jury award. Parents ordered to pay $2 million for not revealing son's HIV. PMID- 15124572 TI - News from the 11th conference on retroviruses and opportunistic infections. PMID- 15124573 TI - A journey toward finding the best medical care. PMID- 15124574 TI - Support groups: tools for living well. PMID- 15124575 TI - Reaching out to the Hispanic community. PMID- 15124576 TI - Lipoatrophy: a new "scarlet letter". PMID- 15124577 TI - Reading food labels. PMID- 15124578 TI - Fear, loathing and discrimination in Las Vegas. PMID- 15124579 TI - Report from the 11th conference on retroviruses and opportunistic infections. Epidemiology. PMID- 15124580 TI - Report from the 11th conference on retroviruses and opportunistic infections. Acute and recent infection. PMID- 15124581 TI - Report from the 11th conference on retroviruses and opportunistic infections. Studies of approved antiretrovirals. PMID- 15124582 TI - Report from the 11th conference on retroviruses and opportunistic infections. STI. PMID- 15124583 TI - Report from the 11th conference on retroviruses and opportunistic infections. Resistance. PMID- 15124584 TI - Report from the 11th conference on retroviruses and opportunistic infections. New drugs. PMID- 15124585 TI - Report from the 11th conference on retroviruses and opportunistic infections. Metabolic complications. PMID- 15124586 TI - Report from the 11th conference on retroviruses and opportunistic infections. Hepatitis. PMID- 15124587 TI - Report from the 11th conference on retroviruses and opportunistic infections. Neurology of HIV. PMID- 15124588 TI - Report from the 11th conference on retroviruses and opportunistic infections. HIV in resource-limited countries. PMID- 15124589 TI - Report from the 11th conference on retroviruses and opportunistic infections. Microbicides. PMID- 15124590 TI - Report from the 11th conference on retroviruses and opportunistic infections. Vaccines and immune-based therapies. PMID- 15124591 TI - Report from the 11th conference on retroviruses and opportunistic infections. OIs and STDs. PMID- 15124592 TI - Grants. HIV/AIDS funding sources still available. PMID- 15124593 TI - International. Rights group: U.S. blocking access to generic AIDS drugs. PMID- 15124594 TI - Drug access. Coalition calls for drugmakers to lower AIDS drug prices. PMID- 15124595 TI - Needlestick injury. HIV-related emotional distress is compensable. PMID- 15124596 TI - Sentencing. Tough sentence for HIV-positive drug dealer upheld. PMID- 15124597 TI - HIV-prevention program fails rape survivors in Africa, group says. PMID- 15124598 TI - Prisons. Inhumane conditions threaten lives of inmates with HIV. PMID- 15124599 TI - Prisons. Doctor offers remedies for HIV ward. PMID- 15124600 TI - SSI benefits denied man who could not return to work. PMID- 15124601 TI - Veteran with AIDS wins disability appeal. PMID- 15124602 TI - Doctor loses license after failing to prove HIV discrimination. PMID- 15124603 TI - Predator label dropped for man who exposed victim to HIV. PMID- 15124604 TI - Discrimination. Employer's word insufficient to dismiss HIV bias case. PMID- 15124605 TI - Suicidal man loses HIV discrimination claim. PMID- 15124606 TI - Massachusetts university to begin human HIV vaccine trials. PMID- 15124607 TI - Gertie Florentine Marx. PMID- 15124608 TI - Margaret Thaler Singer. PMID- 15124610 TI - Views of doctors on clinical correspondence: questionnaire survey and audit of content of letters. PMID- 15124615 TI - Using diazepam and atropine before strabismus surgery to prevent postoperative nausea and vomiting: a randomized, controlled study. PMID- 15124616 TI - AIDS orphans in Africa. PMID- 15124617 TI - The seven habits of highly effective drug discoverers? PMID- 15124618 TI - Rat genome sequence reignites preclinical model debate. PMID- 15124619 TI - Another long leaderless period in store for FDA. PMID- 15124620 TI - Patentability in Europe. PMID- 15124621 TI - Mark C. Fishman. PMID- 15124622 TI - The fast track effect. PMID- 15124623 TI - Tadalafil and vardenafil. PMID- 15124624 TI - [Application of cocaine in maxillofacial surgery]. PMID- 15124625 TI - [State of affairs: forgotten abdominal sponge]. PMID- 15124626 TI - [Bilateral chronic recurrent laryngeal nerve paralysis after bilateral thyroidectomy in follicular thyroid carcinoma (pT1, N0,M0) on the right side]. PMID- 15124627 TI - New inhibitors targeting bacterial RNA polymerase. PMID- 15124628 TI - Prions: so many fibers, so little infectivity. PMID- 15124629 TI - Enzymatic conversion of cytidine to lysidine in anticodon of bacterial isoleucyl tRNA--an alternative way of RNA editing. PMID- 15124630 TI - The PepSY domain: a regulator of peptidase activity in the microbial environment? PMID- 15124631 TI - MANSC: a seven-cysteine-containing domain present in animal membrane and extracellular proteins. PMID- 15124632 TI - Medical slang in British hospitals. AB - The usage, derivation, and psychological, ethical, and legal aspects of slang terminology in medicine are discussed. The colloquial vocabulary is further described and a comprehensive glossary of common UK terms provided in appendix. This forms the first list of slang terms currently in use throughout the British medical establishment. PMID- 15124633 TI - ACOG practice bulletin. Cervical insufficiency. PMID- 15124635 TI - Historical perspectives on the Louisiana State Board of Dentistry. PMID- 15124634 TI - Organized dentistry and dental education: an important partnership. PMID- 15124636 TI - The relationship between periodontal disease and preterm low birth weight and a new Medicaid dental program intervention for pregnant women. PMID- 15124637 TI - An archival perspective of the history of dentistry in Louisiana. PMID- 15124638 TI - Estate planning. PMID- 15124639 TI - A history of dental hygiene in Louisiana. PMID- 15124640 TI - The dental alliances of Louisiana: our history and remembrances. PMID- 15124641 TI - Proceedings of the TRANSCRIPTOME conferences and the SYSTEMOSCOPE International consortium, 2000, 2002, 2003. PMID- 15124642 TI - The fragments of the Methodists: Methodism outside Soranus.. PMID- 15124643 TI - Dennis Chapman. PMID- 15124644 TI - Robert Ernest Davies. PMID- 15124645 TI - Sir John Carew Eccles, A.C. PMID- 15124646 TI - Charles Edmund Ford. PMID- 15124647 TI - Sir John Cowdery Kendrew. PMID- 15124648 TI - Albert Neuberger. PMID- 15124649 TI - Michael Smith. PMID- 15124650 TI - Douglas Robert Wilkie. PMID- 15124651 TI - Olivier Louis Zangwill. PMID- 15124652 TI - Poetical allusions to the circulation of blood up to the end of the seventeenth century. AB - The history of medicine has conventionally been studied by research into prose medical texts, which provide an index of the depth of the professional knowledge available to the contemporary physician, but examination of poetry gives a measure of the diffusion of this knowledge into the laity. William Harvey's monumental discovery of the circulation of the blood in 1628 is now acknowledged as a revolutionary milestone in the history of medicine. It was met however with initial rejection by the majority of his colleagues, and it was not until over twenty years later that Harvey and his discoveries start to be mentioned in poetry. It is interesting to note that the eventual acclaim for his work might well have been based on its correspondence with the rain cycle! In the second half of the century, medical poets start to pay tribute to Harvey's contribution, and a number of his colleagues wrote eulogies to him and his work. PMID- 15124653 TI - Jordi Casals-Ariet. PMID- 15124654 TI - [Embalmers/physicians in ancient Egypt]. AB - On a daily basis, priest embalmers carefully carried out mummification of bodies to give them a certain immortality. Could they also have practised the profession of medicine? PMID- 15124655 TI - Sir Thomas Oliver (1853-1942) and the health of antimony workers. AB - Imported antimony sulphide ore has been processed in the north-east of England on Tyneside since 1864, first at Gateshead and then at Willington Quay, until the process closed in 2000. The health of antimony workers was a concern of Sir Thomas Oliver (1853-1942) who was distinguished in the field of occupational medicine, particularly in connection with diseases due to lead exposure. Antimony appears to have fascinated him but he underestimated its toxic effects on the process workers in concluding that they were healthy and that there were no industrial hygiene problems in the process. Subsequent investigations have presented a much less satisfactory picture although in recent times the factory atmosphere had been transformed so that Oliver's view had probably come true. PMID- 15124656 TI - Periprosthetic fracture treatment. PMID- 15124657 TI - Preservation of knee articular surface with retrograde intramedullary nailing of the femur. PMID- 15124658 TI - Flexion-extension gap changes during total knee arthroplasty: effect of posterior cruciate ligament and posterior osteophytes removal. AB - Flexion and extension gap heights were measured in 50 consecutive primary posterior-stabilized total knee arthroplasties (TKAs) to determine whether posterior cruciate ligament (PCL) release or re-establishment of the posterior condylar recess increased gap width. After PCL release, a slight symmetrical increase was noted in both gaps. In extension, gap width increased on average 1.3 mm and 1 mm in the medial and lateral compartments, respectively. The same pattern was observed in flexion, averaging 1.3 mm medially and 1.3 mm laterally. Another increase in the two gaps was observed after the posterior condylar osteophytes were removed and the posterior recess was re-established. The gaps in extension increased, with respect to the base-line value, on average 1.8 mm medially and 1.8 mm laterally, whereas flexion increased an average 2 mm medially and 2.2 mm laterally. No statistical differences were noted between flexion and extension gaps. No independent differences between the flexion and extension gaps were found in any surgical phase. Posterior cruciate ligament removal and re establishment of posterior condylar recess does not require additional consideration in gap balancing during posterior-stabilized TKA. PMID- 15124659 TI - Viscosupplementation with hylan G-F 20 (Synvisc): pain and mobility observations from 74 consecutive patients. AB - Seventy-four patients (120 knees, 360 injections) referred to an orthopedic surgery clinic by their primary care physician for osteoarthritis knee pain received 3 weekly intra-articular injections of hylan G-F 20 (Synvisc; Genzyme Biosurgery, Ridgefield, NJ). Osteoarthritic knee pain, the ability to perform activities, and the amount of pain medication, physiotherapy, and assistive device use were prospectively evaluated 1 month and 1 year after treatment. The majority of patients reported improved or much improved pain at 1 month (99%) and 1 year (91%) after hylan G-F 20 therapy. Patient activity was similarly improved in the majority of patients at 1 month (99%) and 1 year (69%) following physiotherapy. Of the 120 knees treated with hylan G-F 20, pain relief >6 months was reported for 62% of knees. Use of analgesics, nonsteroidal anti-inflammatory drugs, intra-articular steroids, physiotherapy, and assistive devices was reduced with hylan G-F 20 for 75%-100% of patients at 1 month, and was sustained at 1 year in the majority of patients (78%-94%) for the use of analgesics, steroids, and physiotherapy. Three (4%) patients experienced erythema and irritation at the injection site that resolved within 48 hours and did not interfere with the remaining injections. Hylan G-F 20 effectively and safely relieves osteoarthritis knee pain, facilitates an improved activity level, and decreases the need for pain medication, physiotherapy, and assistive devices. PMID- 15124660 TI - Adolescent ACL injury: treatment considerations. AB - Anterior cruciate ligament (ACL) injury in a skeletally immature patient presents unique treatment challenges. In many cases, conservative treatment with bracing and physical therapy fails, resulting in recurrent instability, pain, swelling, and meniscal and chondral injury. The goal of surgical reconstruction is to recreate ACL stability without causing growth plate arrest, leg-length discrepancy, or angular deformity. Patient characteristics such as skeletal age, Tanner stage, onset of menses, family member height, growth spurt, recent change in foot size, and growth charts can help the surgeon approximate the degree of skeletal maturity and aid in selecting the timing and safest type of reconstruction. Numerous surgical techniques, ranging from an extra-articular reconstruction to intra-articular graft passage without physeal violation to standard transtibial and transfemoral tunnel placement with physeal violation, have been popularized. The majority of existing studies are retrospective case series, describing a particular author's specific technique experience. This article reviews the basic science and clinical literature, presents a treatment algorithm, and provides several case studies. PMID- 15124661 TI - Early ACL reconstruction in combined ACL-MCL injuries. AB - This study reports 18 patients with 19 combined ligament injuries with complete anterior cruciate ligament (ACL) tear and a minimum grade II medial collateral ligament (MCL) tear who underwent early reconstruction of the ACL and nonoperative treatment of the MCL. Inclusion criteria included ACL reconstruction performed within 3 weeks of initial injury, no history of antecedent injury to the ipsilateral knee, and 2-year follow-up data. Associated injuries were noted in 11 patients including 6 isolated lateral meniscal tears, 1 isolated medial meniscal tear, 5 combined meniscal tears, 1 chondral injury, and 1 patellar fracture. Subjective minimum 2-year follow-up yielded a mean Lysholm score of 94.5 and a mean Tegner activity score of 8.4. Serial clinical examinations demonstrated good functional outcomes, range of motion, and strength. No patient experienced ACL graft failure or valgus instability or required subsequent surgery for chondral or meniscal damage. One patient required a second surgery for arthrofibrosis. Clinical and functional outcomes in this study were good with low motion complication rates. Based on our data, early surgical reconstruction of the ACL and nonoperative treatment of the MCL in combined injuries is acceptable and results in excellent clinical and functional outcomes. PMID- 15124662 TI - Bipolar and monopolar radiofrequency treatment of osteoarthritic knee articular cartilage: acute and temporal effects on cartilage compressive stiffness, permeability, cell synthesis, and extracellular matrix composition. AB - The cellular, biochemical, biomechanical, and histologic effects of radiofrequency-generated heat on osteoarthritic cartilage were assessed. Articular cartilage explants (n=240) from 26 patients undergoing total knee arthroplasty were divided based on Outerbridge grade (I or II/III) and randomly assigned to receive no treatment (controls) or monopolar or bipolar radiofrequency at 15 or 30 W. Both potentially beneficial and harmful effects of radiofrequency treatment of articular cartilage were noted. It will be vital to correlate data from in vitro and in vivo study of radiofrequency thermal chondroplasty to determine the clinical usefulness of this technique. PMID- 15124663 TI - Rehabilitation after knee immobilization in octogenarians with patellar fractures. AB - Between January 1990 and December 1999, 14 octogenarians (8 women and 6 men) underwent surgery for comminuted patellar fracture. Average patient age was 83 years (range: 80-88 years). Follow-up ranged from 1-8 years (mean: 3.5 years). Ten of 14 patients were totally independent, whereas 4 patients used a cane for mobilization. Twelve patients had background diseases. Patients underwent operative treatment with tension band wires followed by cast immobilization (knee in approximately 10 degrees of flexion) for 6 weeks. Immediate full weight bearing was initiated in all patients, and intense rehabilitation was performed after cast removal to increase range of motion. Complete union was noted for all fractures. All patients but 1 had an active extension lag of 10 degrees-20 degrees before physiotherapy and maximum flexion was 70 degrees. After physiotherapy, 4 patients regained full active extension and all patients achieved >100 degrees of flexion. Twelve of 14 patients returned to their pre injury functional level. A slight deterioration was noted in 2 patients. Although knee immobilization may cause severe limitation in range of motion, its use in elderly patients followed by intense rehabilitation is advocated and showed good results. PMID- 15124664 TI - Anterior knee pain due to biplanar rotatory malalignment of the femoral component in total knee arthroplasty. Case report. PMID- 15124665 TI - Popliteal artery rupture during total knee replacement in a patient with Monckeberg's sclerosis. Case report. PMID- 15124666 TI - Pharmacodynamic pitfalls in antifungal treatment. PMID- 15124667 TI - Update on the treatment of cerebral aspergillosis. AB - In aspergillus infections of the central nervous system (CNS) the mortality risk usually exceeds 90%. Data from case-reports and a recent retrospective study suggest that neurosurgical interventions, such as abscess resections, stereotactic drainages, or the use of intraventricular catheters, might improve the outcome in CNS aspergillosis. However, there is a lack of clear evidence supporting an extensive neurosurgical management in these patients. A major reason for the devastating prognosis in CNS aspergillosis is a poor penetration of antifungal drugs into the CNS, with the exception of voriconazole. Treatment with voriconazole results in measurable drug levels in the cerebrospinal fluid, which may exceed the minimal inhibitory concentration for aspergillus. Moreover, voriconazole brain tissue levels exceed those measured for other antifungal drugs. In a recent retrospective study, a complete or partial response occurred in 35% of patients who were treated with voriconazole for CNS aspergillosis with a survival rate of 31%. These data support the use of voriconazole in this clinical setting. An intense neurosurgical management and higher doses of voriconazole might further improve the outcome in CNS aspergillosis, but this needs to be evaluated in future studies. PMID- 15124668 TI - Impact of karyotype on treatment outcome in acute myeloid leukemia. AB - Over the course of the last three decades it has become apparent that the majority of cases of acute myeloid leukemia (AML) are characterized by at least one of a variety of recurrent chromosomal abnormalities. Whilst in many instances it remains uncertain as to which abnormalities represent primary events in the pathogenesis of AML, those which provide critical second hits that are required for progression to full blown leukemia or those that are merely markers of the leukemic process, it is nevertheless clear that diagnostic karyotype is a key determinant of outcome in this disease. Indeed there is mounting evidence to support the notion that cytogenetic analysis can serve to identify biologically distinct subsets of AML that demand tailored therapeutic approaches. This underpins the trend towards more widespread adoption of routine cytogenetic and molecular analysis in the characterization of patients with a diagnosis of acute leukemia. A key challenge for the future is to use this information to achieve greater consensus in risk group assignment of AML which will provide a more reliable framework for determining the most appropriate treatment approach for individual patients with this disease. PMID- 15124669 TI - Risk-adapted therapy of AML: the AMLCG experience. AB - Acute myeloid leukemia (AML) is a heterogeneous composition of biologically defined subgroups. Modern trials aim at developing subgroup-specific therapies. The German AML Cooperative Group 1999 trial asks three questions in a randomized factorial design: high-dose vs. standard-dose AraC during induction therapy; G CSF priming vs. no G-CSF priming; and autologous stem cell transplantation vs. maintenance therapy. An interim analysis with 938 patients reveals subgroup specific differences in treatment efficacies. Thus, the application of high-dose AraC during induction results in a superior outcome as compared to standard-dose AraC in patients with unfavorable prognosis but not in other patients. These results underline the need for large comprehensive trials to allow the detection of therapy effects in biologically defined subgroups of AML. PMID- 15124670 TI - Targeted therapy of AML new concepts. PMID- 15124671 TI - Priming with G-CSF in acute myeloid leukemia: preliminary data of the AMLCG. PMID- 15124672 TI - Priming with GM-CSF for acute myelogenous leukemia (AML): GOELAM data. PMID- 15124673 TI - Nuelasta versus G-CSF in solid tumours. PMID- 15124674 TI - Modern diagnostics in chronic myeloproliferative diseases (CMPDs). AB - According to the new WHO classification a group of chronic myeloproliferative diseases (CMPDs) were defined: chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), chronic eosinophilic leukemia and hypereosinophilic syndrome (CEL/HES), polycythemia vera (PV), chronic idiopathic myelofibrosis (with extramedullary hematopoiesis, CIMF), essential thrombocythemia (ET), and so called CMPD/unclassifiable. As clinical features and laboratory findings differ widely between these diseases several diagnostic approaches are mandatory at diagnosis for classification and are needed also for follow up studies, especially for the measurement of minimal residual disease (MRD). We here outline the laboratory set up at diagnosis and during follow up in CMPDs with specific focus on the respective therapeutical consequences. Only by using a comprehensive diagnostic panel including cytomorphology, cytogenetics, and molecular genetic methods establishing the correct diagnosis, optimizing treatment as well as evaluating treatment response is possible in CMPDs today. PMID- 15124675 TI - Imatinib mesylate selectively influences the cellular metabolism of cytarabine in BCR/ABL negative leukemia cell lines and normal CD34+ progenitor cells. AB - STI-571 (Imatinib/Glivec) has been shown to have synergism with various chemotherapeutic agents including cytosine arabinoside (Ara-C) in BCR/ABL positive leukemia cells. The antiproliferative and proapopotic effects of STI-571 in these experiments are mainly explained by its ability to specifically block the fusion-protein BCR/ABL which has a constitutively active tyrosine kinase activity. We investigated the effects of STI-571 in combination with Ara-C on BCR/ABL negative leukemia cell lines and CD34+ hematopoietic progenitor cells in vitro. Raji, HL-60, K562, Kasumi and KG1a leukemia cells and CD34+ cells from healthy donors were incubated with 5-20 microg/ml Ara-C for 5 h alone or in combination with 10 microg/ml STI-571. Intracellular levels of Ara-CTP measured by HPLC were increased 1.5-3 fold in leukemia cells with most promiment effects in HL-60, Kasumi and Raji cells. In HL-60 cells a linear correlation between the concentration of STI-571 (1-10 microg/ml) and the subsequent levels of Ara-CTP was observed. A linear increase of Ara-CTP could be induced by increasing the incubation time with STI-571 from 2-6 h with a ceiling effect after 8 h. In contrast coincubation of mononuclear cells or purified CD34+ cells with STI-571 at therapeutic concentrations lead to decreased intracellular levels of Ara-CTP. The synergism between Ara-C and STI-571 was even more pronounced in Raji and HL 60 cells when 300 ng/ml G-CSF were added at the beginning of the culture period. Intracellular measurements of STI-571 revealed no decreased or increased levels of the compound when increasing Ara-C concentrations were used. Our findings indicate that STI-571 can have significant impact on nucleoside metabolism in malignant and non-malignant hematopoietic cells. Further investigations will have to show whether theses effects can lead to increased cytotoxicity in primary blasts of patients with acute leukemia. PMID- 15124676 TI - Imatinib mesylate (Glivec, Gleevec) in the treatment of chronic myelogenous leukemia (CML) and gastrointestinal stromal tumors (GIST). PMID- 15124678 TI - Transplantation strategies in chronic lymphocytic leukemia: current concepts of the German CLL Study Group and the EBMT. PMID- 15124677 TI - Role of MabCampath in allogeneic transplantation. AB - Alemtuzumab reduces the incidence of acute and chronic GvHD following stem cell transplantation and reduces GvHD-related mortality. There is a delay in immune reconstitution and an increased incidence of viral infections with the use of alemtuzumab, however many of these infections are asymptomatic, and at least in the case of CMV in the sibling setting, do not adversely effect transplant related mortality. Disease relapse appears more common but approaches incorporating DLI may offset this tendency in immune responsive malignancies. Delivery of these therapies in a less toxic manner remains a priority for future research. PMID- 15124680 TI - Novel treatment strategies in follicular lymphoma. AB - Malignant lymphomas are a heterogenous group of malignancies, belonging to the 10 most frequent types of cancers worldwide. In indolent lymphoma only patients with limited stage I/II (Ann Arbor) can be potentially cured by local irradiation. However, about 85% of cases present with advanced stage; for these patients no established therapeutic strategy with curative potential exist. The development of the anti-CD20 monoclonal antibody rituximab has been one of the major contributions in the field of anti-lymphoma treatment in the recent years. Various clinical trials have suggested the combination of chemotherapy and rituximab to be highly effective in de novo as well as relapsed indolent lymphomas. PMID- 15124679 TI - Treosulfan/fludarabine: a new conditioning regimen in allogeneic transplantation. AB - Recently, the water-soluble bifunctional alkylating agent treosulfan demonstrated broad stem cell toxicity, immunosuppressive as well as antileukemic activity. Due to its well known low non-hematologic toxicity profile, treosulfan was considered an alternative agent for conditioning prior to allogeneic transplantation. A first clinical study, combining 3 x 10 g/m2 of treosulfan with 5 x 30 mg/m2 of fludarabine, demonstrated the feasibility of this conditioning. A fast, reliable and complete development of the donor hematopoiesis was evident as well as a low non-hematologic toxicity, transplantation-related mortality and relapse rate. In a second study treosulfan was escalated from 3 x 10 to 3 x 12 and 3 x 14 g/m2. In this protocol, 55 pts (patients) not amenable to standard conditioning suffering from various hematological malignancies were included. Complete donor chimerism was reached by day 28 in 80% of the pts. So far, 8 pts (11%) died without disease progression and 11 pts (20%) relapsed. Treosulfan was very well tolerated. Especially no hepatic VOD, severe cardiac or pulmonary toxicity was noted. Acute GvHD (degrees 11-IV) occurred in 44% and chronic GvHD in 45% of pts. Considering the poor prognosis of these study populations, treosulfan-based conditioning is considered to be safe and efficient. New phase 11 clinical protocols in AML and MDS will be initiated. PMID- 15124681 TI - New treatment strategies in lymphomas: aggressive lymphomas. AB - This review will cover the use of these monoclonal antibodies alone or in combination with chemotherapy for the treatment of aggressive lymphomas. Rituximab, an unconjugated anti-CD20 chimeric antibody, is certainly the most widely used but other unconjugated or radiolabeled monoclonal antibodies may catch up quickly. Rituximab combined with chemotherapy allows increasing the complete response rate, to decrease progression during treatment or relapse, to increase duration of response, event-free survival and overall survival. This benefit is now demonstrated in several randomized studies in different settings. Less data are available for the use of Rituximab in maintenance after chemotherapy or autologous transplant. The use of monoclonal antibodies (MAb) for the treatment of lymphoma patients appeared some 7 years ago and, firstly, they have been developed for so-called 'low-grade' or indolent lymphomas. Murine antibodies have been used with toxin or isotopes attached to them and, in this case, the antibody is used to specifically transport the active agent, often a radionucleide, to lymphoma cells. In the case of unmodified, naked, monoclonal antibodies, such as rituximab, the chimeric human-mouse antibody fixes the antigen on the membrane of lymphoma cells with the murine antibody part and stimulates the immune host mechanisms through the human Fc part. The fact of fixing the antigen on the cell surface may also trigger a cascade of biologic events leading to the cell death through the apoptotic process. PMID- 15124682 TI - Targeted therapies in myeloid leukemias. AB - There is still a compelling need to improve therapeutic outcome in AML. However, during the past several years our understanding of the genetic basis of AML, and the nature of the mutations that contribute to the phenotype, have been elucidated in cell culture and murine models of leukemia. The validation of various mutant leukemogenic gene products has in turn led to the development of an expanding group of molecular targeted therapies that have potential to improve the therapeutic window for treatment of AML. PMID- 15124683 TI - The recent JALSG study for newly diagnosed patients with acute promyelocytic leukemia (APL). AB - Based on the prognostic factors obtained from our previous APL92 study, in the JALSG APL97 study, we intensified chemotherapy for patients with leukocyte counts > or = 3,000/microL and > or = 10,000/microL, also intensified consolidation chemotherapy, and then tested whether further chemotherapy is required in patients with negative RT-PCR for PML/RARalpha after the completion of consolidation therapy. Of 256 presently evaluable patients, 244 (95%) achieved CR. Predicted 5-year EFS is 67% and predicted 5-year overall survival 84%. PMID- 15124684 TI - Monitoring of minimal residual disease in acute promyelocytic leukemia. PMID- 15124685 TI - Curative therapeutic approaches to APL. AB - Acute promyelocytic leukemia (APL) has become the most curable subtype of acute myeloid leukemia in adults. It represents the only established example of successful differentiation therapy. With current therapy which includes all-trans retinoic acid (ATRA) and anthracycline-based chemotherapy for induction, anthracycline-based consolidation and maintenance with ATRA and/or low-dose chemotherapy, approximately 75-85% of patients with acute promyelocytic leukemia (APL) remain alive and disease-free at 5 years, and most patients are likely to be cured, an unprecedented achievement in the field of hematologic malignancies. However, several causes for failure to be cured need to be addressed. The first is early death which occurs in approximately 10% and is frequently attributable to hemorrhage due to the characteristic coagulopathy. The second is relapse, particularly in intermediate- and high-risk patients. Analyses of new prognostic factors may permit refinement of current risk classification and identify patients warranting alternative therapy. Finally, long-term consequences of current treatment will be important to recognize, including delayed cardiomyopathy, extramedullary relapse related to sanctuary sites, and the potential for second malignancies. For patients who do relapse, arsenic trioxide appears to be the treatment of choice since the majority of patients achieve a second complete morphologic, cytogenetic, and even molecular remission. While some patients achieving a second complete remission have prolonged disease-free survival with consolidation and maintenance arsenic, high-dose chemotherapy with autologous hematopoietic stem cell transplantation appears to offer the highest likelihood of cure. Such a strategy or anti-CD33 antibodies, recently shown to be active in APL, might be considered for high-risk patients in first remission. PMID- 15124686 TI - Epigenetic regulation of tumor suppressors in t(8:21)-containing AML. AB - The t(8;21) is perhaps the most frequent chromosomal translocation associated with acute myeloid leukemia. The translocation creates a fusion protein that consists of the DNA binding domain of the RUNX1 transcription factor fused to the MTG8 transcriptional co-repressor to create a potent transcriptional repressor. Here, we discuss the possibility that the t(8;21) fusion protein represses tumor suppressors that regulate the RAS signaling pathway and the p53 oncogenic checkpoint. PMID- 15124688 TI - Farnesyltransferase inhibitors (FTIs) in myeloid malignancies. AB - Farnesyltransferase inhibitors (FTIs) are small-molecule inhibitors that selectively inhibit farnesylation of a number of intracellular substrate proteins such as Ras. Preclinical work has revealed their ability to effectively inhibit tumor growth in vitro and in vivo in animal models across a wide range of malignant phenotypes. Myeloid malignancies are appropriate disease targets, in that they express relevant biologic targets, such as Ras, Mitogen-Activated Protein Kinase (MAPK), AKT, and others that may depend upon farnesyl protein transferase (FTase) activity to promote proliferation and survival. Phase I trials in acute leukemias and myelodysplasia have demonstrated biologic and clinical activities as determined by target enzyme inhibition, low toxicity, and both complete and partial responses. As a result, phase II trials have been initiated in a variety of hematologic malignancies and disease settings, in order to further validate clinical activity and to identify downstream signal transduction targets that may be modified by these agents. It is anticipated that these studies will serve to define the optimal roles of FTIs in patients with hematologic malignancies and provide insight into effective methods by which to combine FTIs with other agents. PMID- 15124687 TI - Core binding factor acute myeloid leukemia. Cancer and Leukemia Group B (CALGB) Study 8461. PMID- 15124689 TI - PKC 412 FLT3 inhibitor therapy in AML: results of a phase II trial. PMID- 15124690 TI - Valproic acid: an old drug newly discovered as inhibitor of histone deacetylases. AB - Fusion proteins encoded by several types of chromosomal translocations in promyelocytic leukemia can serve as aberrant transcriptional repressors relying on recruitment of histonedeacetylases (HDACs) into DNA-associated multi-protein complexes. Thus, inappropriate modulation of chromatin structure by HDACs and subsequently repression of gene expression that is critical for myeloid differentiation appear to be major factors in the development of the disease. They identify inhibitors of HDACs as prime candidates for novel anti leukemic drugs. Over the last years several candidate compounds have been introduced into clinical trials and have successfully been used in compassionate use protocols. Amongst them phenylbutyrate served as the first example to establish proof of principle. Novel drugs such as suberoylanilide hydroxamic acid (SAHA) are developed for example by modifications of the microbial HDAC inhibitory compound trichostatin A with a hydroxamic acid as the key structural element. The branched chain carboxylic acid valproic acid (VPA) that is in use as antiepileptic drug over decades was also discovered to inhibit HDACs and preferentially class I HDACs. HDAC inhibition is likely to mediate the teratogenic side effects of VPA but not the antiepileptic activity. In contrast to other HDAC inhibitors VPA also induces proteasomal degradation of HDAC2. None of the currently available compounds may be the optimum HDAC inhibitory drug but each of them may serve to answer urgent questions concerning the concept of HDAC inhibition in the treatment of malignant diseases. Prominent questions are i) whether and by which mechanisms HDAC inhibition can be expected to affect a malignant disease not only in the early stage but also at later stages that have acquired additional genetic defects, ii) which forms of cancer in addition to myelocytic leukemia respond to HDAC inhibition, iii) by which markers those susceptible forms could be identified and iv) which individual HDACs are the most critical isoenzymes to address in treatment of malignant diseases. PMID- 15124691 TI - G3139, a BCL-2 antisense oligo-nucleotide, in AML. PMID- 15124692 TI - New designs for phase II trials: application to a trial of targeted therapies vs. chemotherapy in patients age > 60 with AML/high-risk MDS. PMID- 15124693 TI - Genetic classification of acute myeloid leukemia (AML). AB - In 50-60% of patients with acute myeloid leukemia (AML) acquired clonal chromosome aberrations can be observed after metaphase banding analyses. The cytogenetic results at diagnosis provide the most single important parameter for determining prognosis so far. Numerous recurrent karyotype abnormalities have been described in AML. These findings on the chromosomal level were followed and supplied by molecular studies that have identified genes involved in leukemogenesis. Even more, molecular markers such as MLL partial tandem duplications (MLL-PTD) or FLT3 length mutations (FLT3-LM) were found to characterize specific subtypes of AML and completed the genetic marker profile. The identification of specific chromosomal abnormalities or molecular markers and their correlation with cytomorphological features, immunophenotype as well as clinical outcome led to a new understanding of AML as a heterogeneous group of distinct biological entities. The importance of cytogenetic and molecular genetic findings in AML for classification and for the understanding of pathogenetic mechanisms is increasingly appreciated in clinical context and was translated also into the new WHO-classification of AML that uses cytogenetic abnormalities as a major criterion. PMID- 15124694 TI - Subgroup specific therapy effects in AML: AMLCG data. PMID- 15124695 TI - P-glycoprotein (Pgp) modulation in untreated acute myeloid leukemia (AML): Cancer and Leukemia Group B (CALGB) trials in younger and older adults. PMID- 15124696 TI - Treatment of acute myeloid leukemia younger adults: the GOELAM experience. PMID- 15124697 TI - Age and the nature of acute myeloid leukemia. PMID- 15124698 TI - MRC trials in childhood acute myeloid leukaemia. AB - The modern approach to therapy for acute myeloid leukaemia (AML) in children began in the late 80's and in the MRC series led to a 30% improvement in survival, up to levels of about 50%. Since 1995 the most recent trial AML 12 has taken those figures to two thirds event free survival and similar overall survival. Resistant disease rates remain at 4% overall but the death rate in complete remission has fallen from 11% to 6% despite increasing intensity of therapy, and due to advances in supportive care including nutrition and antibiotics/antifungals. However, although relapse rates have continued to fall, the biggest challenge is to reduce the currently one third relapse rate. We are much better at predicting who is likely to relapse, based mainly on primary resistance to therapy and karyotype. Analysis of 629 out of the last 808 cases in whom cytogenetic testing was successful (78%) has shown very clearly that t(8;21), t(15;17), inv(16) are independent good risk features. Additionally, loss of a sex chromosome in the 8;21 group defines a group which does exceptionally well, with 93% EFS at 5 years. Chromosome 7 abnormalities also remain of independent prognostic significance when age, WHO classification and white cell count are taken into account, with monosomy 7 doing even worse than 7q abnormalities. The current trial MRC AML 15 investigates the role of fludarabine- idarubicin combination therapy in the induction courses and the role of high dose cytarabine during consolidation; the aim being to increase efficacy and reduce toxicity, particularly that involving the heart. New approaches such as targeted antibody therapy will be explored when toxicity data for children permits. PMID- 15124699 TI - Prognostic relevance of risk groups in the pediatric AML-BFM trials 93 and 98. PMID- 15124700 TI - Maintenance therapy in childhood acute myeloid leukemia. AB - PURPOSE: To determine whether, after very intensive induction and consolidation therapy in childhood AML, further maintenance therapy (MT) confers any advantage. PATIENTS AND METHODS: Three hundred-nine children with previously untreated AML were registered in the LAME 89/91 protocol. This three-cycle intensive regimen included an induction phase (mitoxantrone plus cytarabine) and, for non allografted patients, two consolidation courses, one containing timed-sequential high-dose cytarabine, asparaginase and amsacrine. In the LAME 89 study, patients were given an additional MT consisting of mercaptopurine and cytarabine for 18 months. In the LAME 91 trial, patients were randomized to be given or not MT after consolidation therapy. RESULTS: Out of 309 patients, 276 (90%) achieved a complete remission. The overall survival (OS) and event-free survival at 6 years for all patients were 60% +/- 6% and 48% +/- 6%, respectively. For the complete responders after consolidation therapy, the 5-year OS was significantly better in patients randomized for no further treatment than in patients randomized for MT (81% +/- 13% vs 58% +/- 15%; p = 0.04) whilst the 5-year disease-free survival was not significantly different (60% +/- 19% vs 50% +/- 15%; p = 0.25). The improvement of OS in MT-patients appeared to be related to a higher salvage rate after relapse. CONCLUSION: Over 50% of patients can be cured of AML in childhood. In the context of a very short and drug-intensive regimen, low-dose MT, owing to the lack of improvement in disease control and the worsening of survival, should not be recommended. Over the past 20 years, the outcome of acute myeloid leukemia (AML) in children has improved substantially. In the eighties, complete remission (CR) was achieved in nearly 90% of patients but event-free survival (EFS) was poor. Myeloablative therapy followed by allogenic bone-marrow transplantation (allo BMT) from an HLA-identical sibling was demonstrated, in our experience, to be the treatment of choice for improving DFS in children with AML in first remission. The major issue was how best to maintain complete remission for patients without an HLA sibling donor. Whereas several groups continued to include low-dose MT and others decided to omit it, in 1991, our group undertook a prospective randomized trial (LAME 91 protocol), the main aim of which was to assess the efficacy of MT in addition to an intensive induction and consolidation chemotherapy. The main results have been published previously and are now updated and described in a higher number of patients. PMID- 15124701 TI - Intensified induction therapy for children with AML. PMID- 15124702 TI - Evolution of BFM trials for childhood ALL. AB - Up to 80% of pediatric patients with acute lymphoblastic leukemia (ALL) can be cured if intensive therapy is applied. Severe side effects are encountered in all patients of which, however, only the minority is life-threatening. The leading cause of failure in childhood ALL is still recurrence of disease. To reduce the rate of relapses, but also to limit treatment morbidity, the ALL-BFM group has aimed to improve the risk-adaptation of therapy. The most important addition to clinical factors (e.g. age, WBC, extramedullary involvement), and biological characteristics (such as immunphenotype and cytogenetics), was the recognition of early in vivo treatment response as the strongest predictor for relapse. The determination of leukemic blasts in peripheral blood after exposure to 7 days of prednisone (PRED) and one dose of intrathecal methotrexate (prednisone response) as developed by BFM identified multidrug resistant patients: Such patients had still more than 1,000 blasts per microL at day 8 of therapy (defined as PRED poor responders, 10% of all patients). Prognosis for these was only approximately 35% as compared to approximately 80% in patients with adequate PRED response. Patient characteristics at relapse reveal that most of them were originally comprised in "good risk" patient subgroups: e.g., in trial ALL-BFM 90, 50% of the relapses were noted in patients with c-ALL even though that group had an EFS of 82% (SE 1%). 70% of the recurrences are found among patients with good response to PRED indicating the lack of specificity in the definition of that subgroup. Therefore, the more refined way of determining in vivo response based on the detection of minimal residual disease (MRD) at defined timepoints by identifying clone specific T-cell receptor- (TCR) or immunglobuline (Ig) gene rearrangements appears to be able to define the patient at high risk to relapse more specifically. In the current ALL-BFM strategy, the high sensitivity of the method is utilized to apply treatment reduction in patients with fast clearance of leukemia. Persistent disease in contrast is an indication for treatment modification and intensification. Logistics and quality controls are demanding but essential for the introduction of this new technology into clinical practice. PMID- 15124704 TI - The U.S. trials in adult acute lymphoblastic leukemia. PMID- 15124703 TI - Rationale and design of Total Therapy Study XV for newly diagnosed childhood acute lymphoblastic leukemia. AB - The current cure rate of 80% in childhood acute lymphoblastic leukemia (ALL) attests to the effectiveness of risk-directed therapy developed through well designed clinical trials. The ongoing Total Therapy Study XV at St. Jude Children's Research Hospital was designed to further increase cure rate and to improve quality of life. The study consists of intensive systemic and intrathecal therapy but does not include cranial irradiation, irrespective of a patient's risk features. The intensity of postremission consolidation, continuation and reinduction therapy is based on the level of minimal residual disease at the end of induction, as measured by both flow cytometric detection of aberrant immunophenotypes and polymerase-chain-reaction amplification of clonal antigen receptor gene rearrangements. Status of thiopurine methyltransferase is determined prospectively for treatment modification. Pharmacogenetic, pharmacodynamic, gene expression and proteomic profiling studies of host normal cells and leukemic cells are performed in parallel to elucidate the mechanisms of drug resistance and to advance our understanding of leukemogenesis. PMID- 15124705 TI - Risk/MRD adapted GMALL trials in adult ALL. AB - The German Multicenter Study Group for Adult ALL (GMALL) conducts since 1984 trials with risk adapted study design. The model of conventional prognostic factors comprises now WBC, age, immunophenotype, cytogenetics and molecular genetics. Risk stratification according to these factors allows a highly significant prediction of relapse risk in adult ALL. In the recent GMALL study minimal residual disease (MRD) was added to the risk model. Trials in childhood and adult ALL showed convincingly that MRD is a relevant and independent prognostic factor. It is of particular value in standard risk (SR) patients as defined by conventional factors. In the current GMALL study a risk stratification according to conventional factors is followed by a MRD based stratification in SR patients. Whereas high and very high risk patients receive a stem cell transplantation (SCT) in first CR after induction and first consolidation, SR patients receive cyclic consolidation therapy for one year with MRD monitoring. At the end of the first year a stratification according to course and level of MRD takes place. Treatment is stopped in patients with low risk whereas in high risk patients a SCT is planned. Patients who cannot be allocated to either group are treated as intermediate risk and receive one year of intensified maintenance therapy. Preliminary results show that MRD based risk stratification is feasible and that the treatment recommendations for MRD based risk groups are reasonable. In the future however an earlier identification of high risk patients (after 4 months) will be attempted. PMID- 15124706 TI - Autologous versus allogeneic stem cell transplantation in acute myeloid leukemia. AB - Using the data of the patients in complete remission (CR) up to the age of 45 years included in the EORTC-LG/GIMEMA AML-10 trial we investigated the value of the strategy to perform either an autologous (auto-SCT) or an allogeneic (allo SCT) stem cell transplantation on an intention to treat basis. Between 1993 and 1999, out of 1198 pts, 822 achieved CR. 734 pts, constituting the study group, received an intensieve consolidation course: 293 had a sibling donor and 441 had not. Allo-SCT and auto-SCT was performed in 68.9% and 55.8%, respectively. Cytogenetics was successfully performed in 446 pts. Risk groups were: good (t(8;21), inv(16)), intermediate (NN or -Y only), bad/very bad (all others). Median follow-up was 4 years. The 4-year disease-free survival (DFS) rate of patients with a donor vs of those without a donor was 52.2% vs 42.2%, p = 0.044; the relapse incidence was 30.4% vs 52.5%, death in first complete remission was 17.4% vs 5.3%, and the survival rate was 58.3% vs 50.8% (p = 0.18). The DFS rates in pts with and without a sibling donor were similar in pts with good or intermediate risk cytogenetics, but 43.4% and 18.4%, respectively, in pts with bad or very bad risk cytogenetics. In younger patients (15-35 yrs), the difference was more pronounced. The strategy to perform an allo-SCT in patients where a family donor was available led to better overall results than to perform an auto-SCT, especially for younger patients or those with bad or very bad risk cytogenetics. PMID- 15124707 TI - Transplantation strategies in AML: AMLCG data. AB - Allogeneic stem cell transplantation (allo-SCT) is considered the most potent postremission therapy for acute myeloid leukemia (AML). Its superior antileukemic activity is largely ascribed to the powerful graft-versus-leukemia (GvL) effects exerted by donor lymphocytes. However, due to considerable treatment-related lethality the gains in relapse prevention do not necessarily translate into survival advantages in the overall patient population. Therefore, allo-SCT for adult patients with AML in first complete remission (CR1) is currently recommended only for younger and medically fit patients who are at intermediate to high risk of relapse and have an HLA-identical sibling donor. Stem cell allografting from alternative donors in CR1 is considered an option for high risk patients as defined by cytogenetic abnormalities or incomplete response after one course of induction chemotherapy and should usually be performed in the context of a clinical protocol. PMID- 15124708 TI - Residual sleepiness in patients with optimally treated sleep apnea: a case for hypoxia-induced oxidative brain injury. PMID- 15124709 TI - Sleep disorders and ADHD: shared and common phenotypes. PMID- 15124710 TI - Development of a new manual for characterizing sleep. PMID- 15124711 TI - Long-term intermittent hypoxia in mice: protracted hypersomnolence with oxidative injury to sleep-wake brain regions. AB - STUDY OBJECTIVES: This study was designed to test the hypothesis that long-term intermittent hypoxia (LTIH), modeling the hypoxia-reoxygenation events of sleep apnea, results in oxidative neural injury, including wake-promoting neural groups, and that this injury contributes to residual impaired maintenance of wakefulness. DESIGN: Sleep times and oxidative-injury parameters were compared for mice exposed to LTIH and mice exposed to sham LTIH. SUBJECTS: Adult male C57BL/6J mice were studied. INTERVENTIONS: Mice were exposed to LTIH or sham LTIH in the lights-on period daily for 8 weeks. Electrophysiologic sleep-wake recordings and oxidative-injury measures were performed either immediately or 2 weeks following LTIH exposures. MEASUREMENTS AND RESULTS: At both intervals, total sleep time per 24 hours in LTIH-exposed mice was increased by more than 2 hours, (P<.01). Mean sleep latency was reduced in LTIH-exposed mice relative to sham LTIH mice (8.9 +/- 1.0 minutes vs 12.7 +/- 0.5 minutes, respectively, P<.01). Oxidative injury was present 2 weeks following LTIH in wake-promoting regions of the basal forebrain and brainstem: elevated isoprostane 8,12-iso IPF2alpha-VI, 22%, P<.05; increased protein carbonylation, 50%, P<.05, increased nitration, 200%, P<.05, and induction of antioxidant enzymes glutathione reductase and methionine sulfoxide reductase A, P<.01. CONCLUSIONS: Exposure to LTIH results in an array of significant oxidative injuries in sleep-wake regions of the brain, and these biochemical changes are associated with marked hypersomnolence and increased susceptibility to short-term sleep loss. The residual forebrain redox alterations in wake-promoting brain regions may contribute to persistent sleepiness in a prevalent disorder, obstructive sleep apnea. PMID- 15124713 TI - Sleep deprivation and fatigue in residency training: results of a national survey of first- and second-year residents. AB - STUDY OBJECTIVES: To examine the relationship between residents' self-reported sleep hours, work hours, and other empirical correlates. DESIGN: Using the American Medical Association's Graduate Medical Education database, a national, random sample of PGY (postgraduate year) 1 and PGY2 residents in the 1998-1999 training year was surveyed by mail. MEASUREMENTS AND RESULTS: Residents completed a 5-page survey with 44 questions requiring 144 separate responses about their residency experience. Completed surveys were received from 3,604 of 5,616 residents contacted, a 64.2% response rate. Although work hours and sleep hours were significantly correlated (r = -.39), this relationship was less robust than is generally assumed. Total average sleep hours varied across specialties but also within specialties. Just over 20% of all residents reported sleeping an average of 5 hours or less per night, with 66% averaging 6 hours or less per night. Residents averaging 5 or fewer hours of sleep per night were more likely to report serious accidents or injuries, conflict with other professional staff, use of alcohol, use of medications to stay awake, noticeable weight change, working in an "impaired condition," and having made significant medical errors. CONCLUSIONS: Reduced sleep hours were significantly related to a number of work related, learning, and personal health variables. Capping residents' work hours is unlikely to fully address the sleep deficits and resulting impairments reported by residents. PMID- 15124712 TI - Kindling stimuli delivered at different times in the sleep-wake cycle. AB - STUDY OBJECTIVES: Clinical and experimental observations argue that sleep disturbances are common and coexist in patients with epilepsy. Our previous observations have suggested that neural-immune interactions between corticotropin releasing hormone (CRH) and interleukin-1 (IL-1) are involved in the regulation of physiological sleep-wake behavior. In the present study, we determined the involvement of CRH and IL-1 in the alteration of sleep-wake activities in rats when amygdala kindling was given either at the beginning of the light period (light-onset kindling) or at the beginning of the dark period (dark-onset kindling). DESIGN: The analysis of sleep-wake activities was performed before and after full-blown kindling, and the actions of CRH and IL-1 were tested by pharmacological blockade. SETTING: Neuroscience Laboratory at China Medical University Hospital. PARTICIPANT AND INTERVENTIONS: Male Sprague-Dawley rats were implanted with electroencephalogram (EEG) electrodes and an intracerebroventricular (ICV) guide cannula. Kindling stimuli delivered via a bipolar electrode placing in the right central nucleus of the amygdala. MEASUREMENT AND RESULTS: Amygdala kindling induced diverse effects on sleep. Slow wave sleep (SWS) and rapid eye movement (REM) sleep decreased during the first 12 hour light period when rats were kindled at light onset. When dark-onset kindling was given, SWS increased but REM sleep was not altered during the first 12-hour dark period. After light-onset kindling, the circulating corticosterone concentrations increased and were blocked by ICV administration of the CRH receptor antagonist, astressin or alpha-hCRH. The ICV administration of the CRH antagonist blocked the light-onset kindling-induced decrease of SWS in a dose dependent manner. After dark-onset kindling, IL-1 mRNA expression in the hippocampus and cortex increased. The dark-onset kindling-induced SWS was blocked in a dose-dependent manner by ICV administration of IL-1 receptor antagonist (IL 1 ra). The slow-wave activity during SWS was enhanced regardless of when kindling occurred, but both CRH antagonists and IL-1 ra had little effect on the alteration of slow-wave activity. CONCLUSION: These observations argue that amygdala-kindling-induced sleep-wake alterations are modulated by central increases in CRH or IL-1. PMID- 15124714 TI - Sleep-related fatal vehicle accidents: characteristics of decisions made by multidisciplinary investigation teams. AB - OBJECTIVES: To analyze factors that explain the attribution of crash causes as sleep-related by accident investigators. DESIGN: Analysis of national database of fatal road accidents studied in depth. All nonprofessional nonintoxicated car drivers responsible for a fatal accident from 1991 to 2001 were included (N = 1464). SETTING: Finland, with approximately 5.1 million inhabitants and 2.3 million motor vehicles. PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS: Comprehensive database recorded by multidisciplinary investigation teams, with specific emphasis on the availability of sleep-related driver variables and sleep related causal decisions by teams. RESULTS: Injury severity, age, and marital status of the responsible car driver were related to the proportion of missing data in fatigue-related variables in the database (sleeping time, time awake, lifetime mileage). While there were differences between investigation teams and their activities, a series of logistic regression models showed that the lack of relevant variables in the database did not affect the proportion of accidents attributed to falling asleep (10% of cases) or as having fatigue-related causal factors (an additional 5% of the cases). The accident type (head-on and running off versus other) and road conditions (dry or wet versus icy or snowy pavement) predicted the investigation teams' attribution of sleep-related causes in all models. CONCLUSIONS: Multidisciplinary teams' attribution of sleep-related causal factors were rather stable, comprising 10% to 15% of the cases investigated, independent of the availability of specific sleep-related information. PMID- 15124715 TI - Obstructive sleep-disordered breathing with a dominant cyclic alternating pattern -a recognizable polysomnographic variant with practical clinical implications. AB - OBJECTIVES: To define the clinical and polysomnographic features of a distinct variant of obstructive sleep-disordered breathing that is remarkably mild during rapid eye movement (REM) sleep. DESIGN: Observational study and evaluation of polysomnographic and clinical records. SETTING: American Academy of Sleep Medicine-accredited multidisciplinary sleep disorders center and laboratory. PATIENTS: 35 medication-free subjects with clinical and polysomnographic severe obstructive sleep-disordered breathing selected for dominance of 1 of 2 disordered breathing patterns. INTERVENTIONS: Positive airway pressure titration. MEASUREMENTS AND RESULTS: Nasal pressure was used to score respiratory events. Sleep was scored by both the standard criteria and cyclic alternating pattern (CAP), and the distribution of respiratory events was tabulated and analyzed. A distinct clinical and polysomnographic syndrome emerged, CAP-dominant sleep disordered breathing, characterized by severe relatively short cycle obstructive events during non-REM sleep that were mild in REM sleep. Characteristics include lower body mass index, fewer apneas, and a lower hypoxic burden as reflected by frequency and severity of nocturnal oxygen saturation. During positive pressure titration, a remarkable respiratory instability emerged selectively during CAP, in contrast to stability during REM sleep. This partial treatment failure was associated with persistent clinical symptoms. CONCLUSIONS: This variant of sleep apnea may reflect a dominant component of respiratory instability and periodic breathing coupled with upper-airway obstruction. Its existence questions the conventional practice of calculating global respiratory indexes. Besides positive airway pressure, measures to treat periodic breathing may be required. PMID- 15124716 TI - Relationship between obstructive sleep apnea and diurnal leptin rhythms. AB - STUDY OBJECTIVES: Several studies have reported an association between obstructive sleep apnea and leptin, a hormone that influences satiety and body weight. We evaluated the relationship of leptin levels and the overnight change in levels with sleep apnea. DESIGN: Cross-sectional analysis of data from a prospective cohort study. SETTING: Case Western Reserve University General Clinical Research Center. PATIENTS OR PARTICIPANTS: A total of 138 individuals participating in the Cleveland Family Study--59% women, 45% African-American, with mean apnea-hypopnea index (AHI) of 6.9 (range 0-106)--were studied. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Serum leptin was measured at 10:00 pm to 11:00 pm and at 7:00 am to 8:00 am. Leptin levels in both the morning and evening were positively correlated with apnea-hypopnea index (AHI) in age-, sex-, and race-adjusted analyses (P < .0001) but not after additional adjustment for body mass index. The evening/morning leptin ratio, however, was associated with AHI independent of age, sex, race, body mass index, and waist-hip ratio (P = .03). Conversely, AHI was an independent predictor of the leptin ratio (P = .001) and more predictive than arousal index, oxygenation indices, time asleep, or sleep-stage distribution. An AHI > 15 was associated with a 23% increase in leptin ratio. CONCLUSIONS: These findings suggest that sleep apnea may suppress secretion of leptin in the morning. Alternatively, the relative elevation in evening leptin may influence apnea pathogenesis. PMID- 15124717 TI - Expiratory changes in pressure: flow ratio during sleep in patients with sleep disordered breathing. AB - OBJECTIVES: The size of the upper airway is smallest during sleep, at the end of expiration. This may favor upper-airway collapse in patients with obstructive sleep apnea. In the respiratory cycles preceding obstructive events during sleep, our hypothesis is that upper-airway resistance (UAR) increased earlier during expiration prior to changes occurring during inspiration. DESIGN: We analyzed the pharyngeal pressure-to-flow ratios in order to determine variations in UAR for both inspiration and expiration during stable respiration and the 4 consecutive breaths preceding upper-airway obstructive events in stage 2 sleep. To assess the variation of resistance throughout the within-breath period during stable respiration and the 4 breaths preceding obstructive events, results were expressed as the instantaneous resistance at fixed points 10%, 30%, 50%, 70%, and 90% of time values of inspiration and expiration. Global inspiratory and expiratory UARs during wakefulness and sleep in stable respiration were expressed by the median of instantaneous UAR values. SETTING: Tertiary-care academic medical center. PATIENTS: Eleven patients with moderate to severe sleep disordered breathing. INTERVENTION: None. MEASUREMENTS AND RESULTS: During stable respiration, both inspiratory and expiratory resistance increased during sleep, compared to values while awake. The difference between inspiratory and expiratory UAR increased when sleep deepened. During the respiratory cycle, the increase in the end-expiratory UAR occurred earlier than during inspiration; during stable respiration, UAR was much aggravated during the last three breaths preceding an obstructive event. CONCLUSION: Increases in the expiratory UAR occurred earlier than during inspiration in the cycles preceding upper-airway collapse in patients with sleep apnea. This finding suggested an important role of the expiratory phase in promoting upper-airway collapse and is in accordance with the inspiratory pharyngeal instability occurring when lowering the expiratory pressure in patients with obstructive sleep apnea. PMID- 15124718 TI - Auto-titrating versus standard continuous positive airway pressure for the treatment of obstructive sleep apnea: results of a meta-analysis. AB - STUDY OBJECTIVE: To compare the effectiveness of auto-titrating continuous positive airway pressure (APAP) versus conventional continuous positive airway pressure (CPAP) in reducing the apnea-hypopnea index (AHI), reducing the mean airway pressure, improving subjective sleepiness, and improving treatment adherence in patients with obstructive sleep apnea (OSA). DESIGN: Meta-analysis and metaregression of published randomized trials comparing APAP to CPAP. SETTING: N/A. PARTICIPANTS: N/A. INTERVENTIONS: N/A. RESULTS: We identified 9 randomized trials studying a total of 282 patients. Compared to CPAP, there was no significant advantage of APAP in reducing AHI or sleepiness (pooled APAP-CPAP posttreatment AHI and Epworth Sleepiness Scale score = -0.20 events per hour, 95% confidence interval:[-0.74,0.35], and -0.56 [-1.4,0.3] respectively). The use of APAP reduced the mean applied pressure across the night by 2.2 cm water [1.9,2.5] compared to CPAP. Adherence with therapy was not substantially improved with APAP; pooled estimate of improvement was 0.20 hours per night ([-0.16,0.57], P = .28) using a random-effects model. CONCLUSIONS: Compared to standard CPAP, APAP is associated with a reduction in mean pressure. However, APAP and standard CPAP were similar in adherence and their ability to eliminate respiratory events and to improve subjective sleepiness. Given that APAP is more costly than standard CPAP, APAP should not be considered first-line chronic therapy in all patients with OSA. However, APAP may be useful in other situations (eg, home titrations, detection of mouth leak) or in certain subgroups of patients with OSA. Identifying circumstances in which APAP is a definite improvement over CPAP in terms of costs or effects should be the focus of future studies. PMID- 15124719 TI - Tongue-muscle training by intraoral electrical neurostimulation in patients with obstructive sleep apnea. AB - STUDY OBJECTIVES: To investigate the efficacy of tongue-muscle training by electrical neurostimulation of the upper-airway muscles as an alternative therapy option for obstructive sleep apnea syndrome. DESIGN: A randomized, placebo controlled, double-blind study. SETTING: Department of pneumology and sleep laboratory, University of Witten/Herdecke, Germany. PATIENTS: 67 patients with an apnea-hypopnea index of 10 to 40 per hour were randomly assigned to 2 groups: a treatment group of 33 patients (mean age, 50.8 +/- 12.1 years; mean body mass index, 29.1 +/- 4.4 kg/m2) and a placebo group of 34 patients (mean age, 53.3 +/- 11.3 years; mean body mass index, 28.9 +/- 4.9 kg/m2). Fifty-seven patients completed the study. INTERVENTIONS: Tongue-muscle training during the daytime for 20 minutes twice a day for 8 weeks. MEASUREMENTS AND RESULTS: Treatment efficacy was examined by polysomnography. Snoring, but not apnea-hypopnea index, improved with stimulation (snoring baseline, 63.9 +/- 23.1 epochs per hour; stimulation training, 47.5 +/- 31.2; P < .05) but not with placebo training (snoring baseline, 62.4 +/- 26.1 epochs per hour; placebo, 62.1 +/- 23.8; NS.). CONCLUSIONS: Although tongue-muscle training cannot generally be recommended for the treatment of sleep apnea, the method has proven to be effective in the treatment of snoring. PMID- 15124720 TI - Sleep disorders and daytime sleepiness in children with attention deficit/hyperactive disorder. AB - STUDY OBJECTIVES: Children with attention-deficit/hyperactive disorder (ADHD), in spite of being hyperactive, still benefit from treatment with stimulant medications. We hypothesized that children with ADHD are in fact sleepy during the day, and we sought to test it objectively. DESIGN: Single blind comparative study SETTING: University medical center PARTICIPANTS: Thirty-four children with a previous diagnosis of ADHD (mean age +/- SD, 12.4 +/- 4.6 years) and 32 matched controls (mean age, 12.0 +/- 3.6 years). INTERVENTIONS: N/A. MEASUREMENTS: All participants underwent a full-night polysomnographic study followed by a multiple sleep latency test (MSLT). RESULTS: Sleep latency, total sleep time, and sleep efficiency were comparable between the groups, yet children with ADHD were significantly sleepier during the day than those in the control group (mean MSLT score of 21.9 +/- 5.5 minutes versus 27.9 +/- 2.0 minutes, P < .005). Of the children with ADHD, 17 (50%) had signs of sleep-disordered breathing, compared with 7 of the control group (22%, P < .05). Five of the ADHD group had periodic limb movements during sleep (15%) versus none in the control group. Children without sleep-disordered breathing or periodic limb movements during sleep had the lowest nocturnal sleep efficiency and total sleep time. CONCLUSIONS: We conclude that children with ADHD demonstrate objective daytime somnolence, which may explain the beneficial effects of treatment with stimulant medications. Primary sleep disorders, especially sleep-disordered breathing and periodic limb movement disorder, should be looked for in children with ADHD. PMID- 15124721 TI - Sleep and neurobehavioral functioning in boys with attention deficit/hyperactivity disorder and no reported breathing problems. AB - STUDY OBJECTIVE: To investigate by objective measures the neurobehavioral correlates of sleep patterns in boys with attention-deficit/hyperactivity disorder (ADHD) and no reported breathing problems and in control children. DESIGN: Sleep assessment was conducted for 5 consecutive days and neurobehavioral assessment was conducted in the morning of the third day of measurement in children with ADHD and no breathing problems and control children. SETTING: The neurobehavioral assessment was conducted in the school; sleep was assessed in the home. PARTICIPANTS: 25 controls and 24 boys diagnosed with ADHD between 7 and 11 years of age. MEASUREMENTS: Sleep was monitored using actigraphy for 5 consecutive nights. A computerized neurobehavioral evaluation system was used to assess children's neurobehavioral functioning. RESULTS: Canonical correlation analyses revealed different patterns of associations between sleep and neurobehavioral functioning for the 2 groups. Significant relationships were found between measures of sleep and performance on complicated neurobehavioral tasks in the control group but not in the ADHD group. In addition, children with ADHD had increased instability of sleep parameters compared to the controls. CONCLUSIONS: The findings highlighted the distinct nature of associations between sleep and neurobehavioral functioning in boys with ADHD children and no reported breathing problems compared to controls. Despite the similarity between symptoms caused by sleep fragmentation and symptoms of ADHD, these appear to be different clinical conditions. The source of inattentiveness in children should be identified for best planning of clinical intervention. PMID- 15124722 TI - Sleep pressure score: a new index of sleep disruption in snoring children. AB - STUDY OBJECTIVES: Excessive daytime sleepiness (EDS), as measured by objective criteria, is infrequent in snoring children despite a high prevalence of EDS related behavioral manifestations. We hypothesized that sleep architecture and arousal indexes may be altered relative to the severity of sleep-disordered breathing (SDB). DESIGN: Retrospective and prospective study. SETTING: Questionnaires were distributed through sleep clinic or school program; polysomnograms were performed at Kosair Children's Hospital in Louisville, Kentucky. PARTICIPANTS: To examine this issue, 182 children with SDB, 163 children with primary snoring, and 214 control children with a mean age of 6.9 +/ 2.6 years underwent polysomnographic evaluation in the laboratory. MEASUREMENTS AND RESULTS: Significant increases in slow-wave sleep (percentage of total sleep time) and decreases in rapid eye movement sleep (percentage of total sleep time) occurred in the SDB group (P < .0001). Spontaneous and respiratory arousal indexes and the apnea-hypopnea index (AHI) displayed negative and positive correlations, respectively, suggesting reciprocal interactions. Based on these observations, a sleep pressure score (SPS) was derived as a surrogate numeric measure for disrupted sleep homeostasis. The SPS exhibited linear increases relative to AHI, reaching a plateau at an AHI of 30 to 40 per hour of total sleep time. Furthermore, SPS values were significantly higher among African American and obese children (P < .0001). CONCLUSIONS: Sleep architecture is not preserved in children with SDB. An algorithm allowing for calculation of sleep propensity and disturbed sleep homeostasis in children who snore is proposed and may be of practical value in the assessment of sleepiness. PMID- 15124723 TI - Sleep pressure correlates of cognitive and behavioral morbidity in snoring children. AB - STUDY OBJECTIVES: Sleep architecture is not preserved in children with sleep disordered breathing but, rather, undergoes dynamic changes that exhibit significant correlation with severity of sleep-disordered breathing. A sleep pressure score (SPS) with a cutoff value of 0.25 was derived from analysis of a large cohort of snoring and control children. Neurocognitive batteries were applied to examine the potential effect of SPS. DESIGN: Prospective study. PARTICIPANTS: 199 children who underwent a battery of neurobehavioral tests following an overnight sleep study were assigned to SPSHigh (> or = 0.25) or SPSLow (< 0.25) groups, and their neurocognitive performances were compared. RESULTS: Children in the SPSHigh group were significantly more likely to have deficits in memory, language abilities, verbal abilities, and some visuospatial functions than were children in the SPSLow group. These effects remained highly significant after adjusting for confounding variables and exhibited small to moderate effect sizes. CONCLUSIONS: We conclude that a sleep-pressure numerical factor derived from the overnight polysomnogram in snoring children is associated with deficits in neurobehavioral daytime functions that is independent of respiratory disturbance and hypoxemia and suggests a significant role for disturbed sleep homeostasis in pediatric sleep-disordered breathing. PMID- 15124724 TI - Vulnerability to stress-related sleep disturbance and hyperarousal. AB - STUDY OBJECTIVES: To determine the presence of a hypothesized trait vulnerability to sleep disturbance and hyperarousal. DESIGN: Polysomnographic assessment of sleep in response to stress during a first night in the laboratory and subsequent physiologic arousal. PARTICIPANTS: One hundred and four individuals (46% men, mean age 40.4 +/- 12.9 years) drawn from a population-based sample. INTERVENTIONS: Individuals were exposed to a first night in the laboratory. MEASUREMENTS AND RESULTS: Participants completed a Likert-scale questionnaire, consisting of 27 items, that assesses sleep disturbance in response to commonly experienced stressful situations. Factor analytic techniques identified a single 9-item factor that was representative of the construct of "stress-related" vulnerability to sleep disturbance. Reliability of the resulting 9-item scale was high (Cronbach's alpha = .83). Individuals with higher scores on this scale, the Ford Insomnia Response to Stress Test (FIRST; median split), had a lower sleep efficiency (P = .001), as well as an increased latency to stage 1 sleep (P = .001) and persistent sleep (P = .002) on the first night of nocturnal polysomnography. Moreover, these high-scoring individuals showed increased arousal as evidenced by an elevated sleep latency on the Multiple Sleep Latency Test compared to individuals with low FIRST scores. Importantly, after controlling for current and past insomnia, the differences between individuals scoring high and low on the FIRST in terms of nocturnal sleep and daytime arousal remained significant. Other stages of sleep (stage 2, slow-wave, and rapid eye movement sleep) were not different between the groups. CONCLUSIONS: These results showing a relationship between FIRST scores and nocturnal polysomnography and Multiple Sleep Latency Test scores have 3 potential implications: (1) the data demonstrate a characteristic that relates to vulnerability to stress-related sleep disturbance as manifested by a first night in the laboratory; (2) the elevated latencies on the Multiple Sleep Latency Test in these individuals, despite significantly disturbed sleep, support the notion of physiologic hyperarousal in these individuals and suggests they may be predisposed to developing chronic primary insomnia; and (3) the vulnerability identified may underlie vulnerability to transient sleep disturbance associated with other sleep disruptive factors. PMID- 15124725 TI - Sleep and reported daytime sleepiness in normal subjects: the Sleep Heart Health Study. AB - STUDY OBJECTIVES: To describe the distribution of nocturnal sleep characteristics and reports of daytime sleepiness in a large well-defined group of healthy adults. DESIGN: The Sleep Heart Health Study is a multicenter study examining sleep and cardiopulmonary parameters through nocturnal polysomnography in adults enrolled in geographically distinct cardiovascular cohorts. SETTING: Community setting. PARTICIPANTS: 470 subjects enrolled in the Sleep Heart Health Study (n = 6440) were selected as a 'normative' group based on screening of health conditions and daily habits that could interfere with sleep. MEASUREMENTS AND RESULTS: Home-based nocturnal polysomnography was obtained on all participants and centrally scored for sleep and respiratory parameters. Demographic and health related data were obtained and updated at the time of the home visit. Sleep efficiency decreased by 1.6% for each 10 years of increased age. Sleep time decreased by 0.1 hours (6.0 minutes) for each 10-year age increase and was longer in women. The arousal index increased by 0.8 for each 10-year increase in age and was lower by 1.4 in women. Women had a lower mean percentage of stage 1 and stage 2 sleep. Mean percentage of slow-wave sleep was higher in women (by 6.7%). Percentage of slow-wave sleep decreased with increased age for men only (by 1.9% for each 10-year age change). CONCLUSIONS: Data suggest a clear lessening in the quantity and quality of sleep with age that appears to be more rapid in males compared to females. PMID- 15124726 TI - Similarities and differences in sleep-wake patterns among adults and their children. AB - STUDY OBJECTIVE: To determine associations and differences in sleep-wake patterns among children and their parents and to explore the correlates for sleep-wake patterns among adults. DESIGN: A school-based cross-sectional survey. PARTICIPANTS: Sample included 1479 fourth to eighth graders and their parents, using a multistage sampling method. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Students and their parents completed a Sleep Habit Questionnaire, which included sleep schedules, a mood scale, and the morningness/eveningness scale. Statistical methods included Pearson and Spearman correlations and analysis of variance using a mixed model. Results showed low correlations in sleep schedules and sleep-wake patterns between children and those parents who did not work shifts or have an evening or night job. Compared to their children, parents tended to go to bed and rise later, have shorter nighttime sleep duration, have less weekend compensation of sleep, and have the morning-type sleep profile. In addition, junior-high students (seventh and eight graders) demonstrated different sleep-wake patterns compared to elementary-school students (fourth to sixth graders). Young age, moodiness, and shift work were associated with tendencies to be the evening type among parent participants. CONCLUSIONS: Our findings indicate that age, social demands (school work for children and adolescents and employment type for adults), and mood status are associated with sleep-wake patterns. Future study examining the association between evening-sleep type and neurotic traits and psychopathology in the adult population will be our next step. PMID- 15124727 TI - Associations between gender and measures of daytime somnolence in the Sleep Heart Health Study. AB - STUDY OBJECTIVES: To examine the relationship of gender to subjective measures of sleepiness, including the Epworth Sleepiness Scale (ESS), in a community-based population. DESIGN: A cross-sectional study. SETTING/PARTICIPANTS: Multicenter Sleep Heart Health Study participants (N = 6.440, 52% women) recruited from ongoing cohort studies. INTERVENTIONS: N/A. MEASUREMENTS: Scores from the ESS, Sleep Heart Health Study daytime sleepiness and feeling unrested questions, polysomnography results (respiratory disturbance index at 4% desaturation), as well as data on difficulty initiating and maintaining sleep, insufficient sleep, sedative use, alcohol use, cardiovascular or respiratory disease, frequent awakening due to leg cramps. RESULTS: Women reported feeling sleepy as often as men did (odds ratio [OR] = 1.06; confidence interval [CI], 0.86-1.32), but women were less likely to have an ESS score > 10 (adjusted OR = 0.77; CI, 0.66-0.90) and more likely to report feeling unrested (adjusted OR = 1.39; CI, 1.14-1.69) than men. In men, the ESS score was more strongly correlated with reports of feeling unrested or sleepy compared to women. CONCLUSIONS: Men and women answer questions on sleepiness differently. Findings indicate that using the ESS to detect subjective sleepiness is more likely to identify men with sleepiness. Since the ESS is more strongly related to other subjective measures in men, the ESS may be a more sensitive measure of subjective sleepiness in men than in women. PMID- 15124728 TI - Periodic limb movements in children: prevalence in a referred population. AB - STUDY OBJECTIVES: To determine the prevalence of periodic limb movements of sleep (PLMS) in children referred for evaluation of sleep disorders. DESIGN: A retrospective analysis of all overnight polysomnograms (PSG) performed at a tertiary-level pediatric care facility. SETTING: All PSG studies were performed in the pediatric sleep laboratory associated with the Alberta Children's Hospital in Calgary, AB, Canada. The pediatric sleep laboratory was situated in the Foothills Hospital Alberta Lung Association Sleep Center until August 2002. At that time, the pediatric sleep laboratory was relocated to the Alberta Children's Hospital. PARTICIPANTS: All children of any age undergoing PSG for any reason between October 1999 and March 2003. MEASUREMENTS AND RESULTS: All PSG records were reviewed, and the following data was extracted: periodic limb movement index (PLM index), periodic limb movements with arousal index (PLM w/arousal index), apnea-hypopnea index (AHI), and patient demographics (age, sex, comorbidities). A total of 591 PSG studies were reviewed. Thirty-three of the 591 children (5.6%) had evidence of PLMs > 5 per hour. Twenty of the 33 (60.0%) had coexistent obstructive sleep apnea (AHI > 1/hour). Only 7 of the 591 children studied (1.2%) had evidence of PLM > 5 per hour with no other comorbidity. Two of 13 children with PLM > 5 per hour and no evidence of obstructive sleep apnea had attention deficit/hyperactivity disorder. The prevalence of PLMS in the 28 of the 591 subjects with a preexisting diagnosis of ADHD was increased at 7.1%. CONCLUSIONS: PLMS is an uncommon disorder of childhood. In a select population at increased risk for having a sleep disorder, the prevalence of isolated PLMS is only 1.2%. PMID- 15124729 TI - Serotonergic antidepressants are associated with REM sleep without atonia. AB - STUDY OBJECTIVES: Rapid eye movement (REM) sleep behavior disorder (RBD) is generally observed in older men and in individuals with specific neurologic diseases. There are case reports of RBD in individuals taking serotonergic antidepressants. Our objective was to assess electromyogram (EMG) activity during REM sleep in individuals taking serotonergic antidepressants and in a matched control group not on such medication. DESIGN: Chart review of clinical and polysomnographic data. SETTING: Sleep laboratory affiliated with a general hospital. PARTICIPANTS: 15 subjects taking a serotonergic antidepressant and 15 age-matched individuals not on such medication. MEASUREMENTS: Submental and anterior tibialis tonic and phasic EMG activity during REM sleep, REM latency, time in REM, apnea-hypopnea index, periodic leg movements of sleep index, and sleep-architecture measures. RESULTS: Tonic, but not phasic, submental EMG activity during REM sleep was significantly more common in the antidepressant treated group than in the control group (P < .02). Tonic REM submental EMG activity correlated with REM latency (r = .42, P = .02) and inversely with REM time (r = -.36, P = .05). Subject age correlated with tonic REM submental EMG activity (r = .58, P = .02) in the antidepressant group There were also trends for more phasic activity in the anterior tibialis (P = .09) and submental (P = .07) EMG in REM sleep in the antidepressant group than in the control group. CONCLUSIONS: Subjects taking serotonergic antidepressants had more EMG activity in the submental lead during REM sleep than did controls. This correlated with measures of REM suppression and age. Individuals taking such medications may be at increased risk of developing REM sleep behavior disorder, particularly with increasing age. PMID- 15124730 TI - An automatic method for scoring leg movements in polygraphic sleep recordings and its validity in comparison to visual scoring. AB - STUDY OBJECTIVES: The study presents an automatic method for scoring leg movements in polysomnographic recordings and describes an empirical investigation of its validity. DESIGN: Leg movements measured by means of the surface electromyogram activation of the right and left tibialis anterior muscle contained in 24 digitally recorded all-night polysomnograms were analyzed visually according to the American Sleep Disorders Association guidelines by 2 experienced raters and automatically scored using a newly developed electromyogram-based analytical method. Two visual scorings and the automatic scoring were compared in pairs using descriptive and confirmative statistical methods. SETTING: N/A. PARTICIPANTS: All-night polysomnograms of adaptation nights at the sleep laboratory of 10 patients with idiopathic restless legs syndrome (RLS) according to the International RLS Study Group. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Agreement rates between the 2 well-trained scorers and the automatic method were comparable. Based on the first scorer's results (100%) 92.5% of the movements were detected by the second scorer and 94.3% by the automatic method. When the visual scorings were compared, the rate of false positive and false-negative errors were 7% and 3%, respectively. Comparing both visual scorings with the results of the automatic scoring yielded false-positive and false-negative rates in the range from 3% to 8%. CONCLUSIONS: The degree of accordance between the 2 visual scorings and between the visual and the automatic scorings were comparable. Therefore, this method is valid and may be used for the automatic detection of leg movements in future studies. PMID- 15124731 TI - A standard procedure enhances the correlation between subjective and objective measures of sleepiness. AB - OBJECTIVE: The goal of this study was to assess whether instituting a standard procedure to minimize transient activation prior to the subjective rating of sleepiness can improve the predictive value of the rating process. METHODS: Thirty young adults, aged 19 to 26 years, participated in the study. Subsequent to sleeping at home with bedtime restricted to 5 hours, they came to the sleep laboratory. They were instructed to rate their level of sleepiness on the Stanford Sleepiness Scale (SSS) and visual analog scales (VAS). A "calm-down" procedure, sitting quietly with eyes closed for 1 minute, was instituted prior to sleepiness ratings for half of the subjects (experimental group) but not for the other half of the subjects (control group). A nap trial with polysomnographic recording was then conducted, followed by a vigilance test. RESULTS: For the experimental group, VAS results of "sleepiness" and "alertness" both correlated significantly with sleep-onset latency during the nap (SOL: r = -.62 and .64, respectively, P values < .05) and with reaction time (RT) on the vigilance test (r = .56 and -.54, P values < .05). The SSS ratings showed significant correlation with nap SOL (r = -.58, P < .05) but not with RT on the vigilance test (r = .19, p = .52). For the control group, none of the subjective ratings showed significant correlation with objective measures. The differences between the resultant correlations for the 2 groups were statistically significant for 2 sets of correlations: the correlation between VAS of "alertness" and nap SOL and the correlation between VAS of "sleepiness" and RT on the vigilance test. CONCLUSION: The results indicate that the subjective ratings of the sleepiness state for individuals with mild sleep restriction more faithfully reflect a physiologic tendency to fall asleep as well as cognitive attentiveness when the ratings are conducted subsequent to sitting still with eyes closed for a sufficient time to minimize transient activation. PMID- 15124732 TI - Why a fly? Using Drosophila to understand the genetics of circadian rhythms and sleep. AB - Among simple model systems, Drosophila has specific advantages for neurobehavioral investigations. It has been particularly useful for understanding the molecular basis of circadian rhythms. In addition, the genetics of fruit-fly sleep are beginning to develop. This review summarizes the current state of understanding of circadian rhythms and sleep in the fruit fly for the readers of Sleep. We note where information is available in mammals, for comparison with findings in fruit flies, to provide an evolutionary perspective, and we focus on recent findings and new questions. We propose that sleep-specific neural activity may alter cellular function and thus accomplish the restorative function or functions of sleep. In conclusion, we sound some cautionary notes about some of the complexities of working with this "simple" organism. PMID- 15124733 TI - Is Eszopiclone appropriate and effective for the long-term clinical management of chronic insomnia? PMID- 15124734 TI - Restless legs syndrome among pregnant women. PMID- 15124735 TI - The upper airway and obstructive sleep apnea in morbidly obese women. AB - STUDY OBJECTIVES: We are currently carrying out a study on the changes of the upper airway in morbidly obese women and the relationship of the upper airway with the severity of obstructive sleep apnea syndrome (OSAS). This study is similar to the study by Fogel RB et al, which appeared in Sleep 2003;2:150-5. DESIGN: Patients underwent a cardiorespiratory polygraphic sleep study, respiratory function test (spirometry, plethysmography, arterial blood gas analysis), and computed tomographic studies of the upper airway. PATIENTS: Thirteen morbidly obese women being evaluated for weight-reduction surgery. RESULTS: OSAS was present in all the patients. We found a positive correlation between the uvula diameter and age (r = 0.63, P = .02) and a negative correlation between the area of the oropharynx at the end of maximal expiration and the number of desaturations greater than 4% per hour (r = -0.58, P = .03). In the subgroup of patients with severe OSAS (7 cases), the area of the oropharynx at maximal inspiration had a negative correlation with the apnea-hypopnea index (r = -0.78, P = .03). The resistances of the airway positively correlated with apnea hypopnea index (r = 0.89, P = .003), apnea index (r = 0.90, P = .03), and desaturation index (r = 0.91, P = .02). The Pao2 had a negative correlation with apnea-hypopnea index (r = -0.63, P = .02), apnea index (r = -0.65, P = .02), and desaturation index (r = -0.66, P = .02). CONCLUSIONS: Our results confirm that, in morbid obese women, airway resistance is higher in those patients with OSAS. We propose that airway resistance and Pao2 can also constitute a good predictor of apnea severity and that a reduction in the cross-sectional area of the airway at the level of the nasopharynx could be related to the severity of OSAS. PMID- 15124736 TI - EMTALA/COBRA. Emergency Medical Treatment and Active Labor Act. Consolidated Omnibus Budget Reconciliation Act of 1985. PMID- 15124737 TI - Speak out against physician assisted suicide and physician assisted death. PMID- 15124738 TI - America's health care mess (dirty little secrets few talk about). PMID- 15124739 TI - The cockroach as a host for Trichinella and Enterobius vermicularis: implications for public health. AB - Cockroaches are known carriers of bacteria and fungi that produce disease in humans. However, the link between pathogenic helminths and cockroaches has not been fully explored. This preliminary study demonstrates Trichinella and Enterobius (also known as human "pinworm") infestation in cockroaches obtained from a grade school and hospitals in Hawaii. This is the first report of Trichinella and Enterobius infestation in naturally occurring cockroaches. These results suggest that roachs are an unappreciated hosts for these human pathogens and are potential reservoirs for these nematodes, supporting their persistence and transmissibility in the environment. Given the ubiquitous nature of the cockroach and the human-occupied settings in which the infested roaches were found, public health concerns are apparent. Furthermore, this information may have diagnostic value when examining Trichinella and Enterobius-infected individuals. PMID- 15124741 TI - Unusual botfly skin infestation. AB - Myiasis, the infestation of humans and animals with fly larvae, is observed in tropical, lowland areas. Dermatobia hominis is a common cause of cutaneous human infestation in these areas. Patients often present with a furuncular lesion on the extremities, back, or scalp. We report a case of furuncular myiasis in a patient returning from a trip to South America. We will discuss the life-cycle of D. hominis and the clinical findings important in the diagnosis of myiasis. PMID- 15124740 TI - Ophthalmomyiasis in Hawaii. AB - Ophthalmomyiasis is the infestation of the eye by fly larvae. Commonly caused by Oestrus ovis, a female sheep botfly will accidentally deposit her larvae into a human eye, resulting in disease. Prompt recognition and treatment of this condition will improve patient care and reduce potential complications. We report a case of ophthalmomyiasis in a young man from Molokai who was infested while unloading a Christmas tree. PMID- 15124742 TI - Fatal postoperative arrhythmia in a man with a remote history of methamphetamine and cocaine use: a case report. AB - Long-term methamphetamine and cocaine use are associated with significant cardiovascular consequences. Despite these demonstrated associations, adverse effects that persist after cessation of drug use are difficult to establish. Cardiovascular pathology associated with long-term drug use may have subclinical presentations that persist long after cessation of drug use. In asymptomatic persons with a remote history of long-term methamphetamine or cocaine use, it may be prudent to assume existing subclinical cardiovascular pathology with positive methamphetamine or cocaine use history as a cardiac risk factor. PMID- 15124743 TI - Melanoma and Hawaii's youth. AB - Hawaii's sandy beaches, warm crystal waters, and mild climate attract tourists and residents alike to enjoy hours of outdoor activities under the sun. As frequent participants of these sun related activities, Hawaii's youth are exposed to high levels and duration of ultraviolet radiation throughout their early lives. This study aims to define occurrence trends of cutaneous malignant melanoma in Hawaii in correlation to increased childhood ultraviolet exposure. This paper addresses trends in melanoma incidence during 1979-2002 for Hawaii residents < 25 years of age. Data obtained from this review were analyzed by age group and ethnicity. Results show that although the incidence of melanoma is increasing for Hawaii residents over 25 years of age, the rate of melanoma occurrence in Hawaii's youth (< 25 years) is not increasing. PMID- 15124744 TI - Students teaching students: Community Health's School Health Education Program (SHEP). PMID- 15124745 TI - Overweight and obesity, a worsening health problem particularly in children. PMID- 15124746 TI - Basic EMTALA requirements. PMID- 15124747 TI - Hepatitis C virus infection at Sharkia Governorate, Egypt: seroprevalence and associated risk factors. AB - Because many persons with chronic hepatitis C virus (HCV) infection are asymptomatic, population based serologic studies are needed to estimate the prevalence of infection and to develop and evaluate prevention efforts. A sample of 1422 individuals was included in the study by using multistage sampling technique. Their age ranged from 4-78 years with a mean age (34.7 +/- 18.5), 782 were males (55%) and 640 were females (45%). Exposures and demographic characteristics were obtained through a predesigned questionnaire. Antibody to HCV was assessed using micro-particle enzyme immunoassay (MEIA) enzyme assay by IMX, and the HCV RNA was tested by Real-time PCR technique using ABI Prism 7700 system. The seroprevalence of antibodies to HCV were 23.4% and 27.4% in urban and rural areas respectively, with an overall prevalence (25.8%). This reflects prior HCV infection but not necessarily a current liver disease. Prevalence was higher among males than females and increased sharply with age, from 4.8% in those < 20 years old to (41.9%) in older ages (> or = 40 years). Those who were not educated and farmers had a significantly high prevalence. The significant predictors of HCV infection were previous parenteral therapy for schistosomiasis (OR = 4.3, 95% CI = 3.6-7.9), among those over 20 years of age (3.5, 2.18-5.8), blood transfusion (4.1, 2.4-6.9), invasive procedures (surgery and endoscopy), and use of contaminated syringes and needles. Also, shaving at community barbers added significance to the model. Exposures not significantly related to HCV seropositivity were gender, active infection with Schistosoma mansoni, sutures or intravenous and urinary catheterization, water pipe "goza" smoking in group. PMID- 15124748 TI - New concept in histopathological grading and staging of chronic hepatitis C infection at Sharkia Governorate, Egypt. AB - Hepatitis C virus (HCV) has been estimated by the WHO to infect 170 million patients worldwide, with a high prevalence rate (about 24.5%) among Egyptians. The disease could be presented with variable hepatic lesions ranging from mild inflammation, fibrosis, cirrhosis to even end stage liver disease and hepatocellular carcinoma. The Knodell histology activity index, published in 1981, was the first system of its type and is widely regarded as the benchmark for objective, semi-quantitative reproducible description of the various morphological lesions of chronic hepatitis. Other proposals for semi-quantitative evaluation have followed. In this study, when applying these systems on the present cases (109 liver biopsies taken from Egyptian patients infected with HCV), the authors found that the presented histopathological features may be unusual for any of the known scoring systems. Therefore, they suggested a new system for grading and staging of liver diseases in Egyptian patients infected with HCV. Accordingly, the degrees of necroinflammations are classified into 3 grades (1-3) and the progression of fibrosis is classified into 3 stages (1-3). The reduced numbers of grades and stages proposed in this study may be attributed to the rapid course among Egyptians who differ in environmental circumstances from abroad. PMID- 15124749 TI - Molecular cytogentic profiles of hepatitis C infection in patients at Sharkia Governorate, Egypt. AB - It has become apparent that hepatitis C virus (HCV) infection is a major risk factor for the development of hepatocellular carcinoma (HCC) worldwide. The precise mechanism by which HCV causes HCC is not known. Unlike the hepatitis B virus (HBV), HCV is not a DNA virus and does not become integrated within the genome of hepatocytes. It is more likely that HCC occurs against a background of inflammation and regeneration, associated with liver injury due to chronic hepatitis. In this study, 40 of paraffin blocks liver tissues from HCV-PCR positive patients (HBV seronegative) were examined using DNA image cytometry to evaluate its role in diagnosing HCC associated with HCV infection. Fluorescent in situ hybridization (FISH) technique using LSIZNF 217 chromosome 20q 13.2 probe was applied as well. The results showed high percentage of S-phase fraction in cases of G2S2 and G3S3 with DNA diploidy. Only two cases of G3S3 showed DNA aneuploidy with severe amplification of chromosome 20q 13.2. Consequently, DNA imaging cytometry is a good approach in differentiating dysplasia from well differentiated HCC on top of HCV infection. In conclusion HCV has an acquired role in development of HCC through amplification of the aggressive tumor behavior oncogene LSIZNF 217 at chromosome 20q 13.2. PMID- 15124750 TI - Haematologial manifestations in HCV infected patients at Sharkia Governerate, Egypt. AB - Hepatitis C virus (HCV) is a major cause of morbidity worldwide. The aim of this study is to evaluate hematological manifestations occurring in patients suffering from chronic HCV infection. Positive HCV-RNA cases (109) were subjected to the following C.B.C., P.T., P.T.T., B.T., C.T., detection of F.D.Ps., measurement of plasma AAT then B.M. aspiration and examination for 20 cases of them. Patients were classified into 3 groups according to the histopathological staging and grading of liver biopsy. Comparison between groups according to histopathological grading and staging for hematological and chemical parameters revealed significant statistical difference in platelets count, S. Albumin, ALT and AST levels. Comparison between groups according to histopathological grading and staging for coagulation profile, AAT level and FDPs revealed significant statistical difference regarding all parameters. Bone marrow aspiration and examination revealed mild hypocellularity with increased number of lymphocytes and relevance of plasmacytoid-lymphocytes. From this study we can conclude that patients with chronic HCV infection are in need for good observation and follow up before taking therapy because they have some hematological abnormalities which need more concern in order to decrease their progressive effect before starting therapy for HCV per se. They should be always screened and given liver and marrow supportive supplements. PMID- 15124751 TI - Autoantibodies in HCV infected patients at Sharkia Governerate, Egypt. AB - It is not clear whether HCV induces an autoimmune disease in infected patients or not. The aim of this study is to evaluate some immunological manifestations in chronic heapatitis C patients and to find out its relationship to liver pathology. The study included 109 positive HCV-RNA patients. They were classified according to liver histopathology into three groups: Group I included 22 patients (G1S1), Group II included 67 patients (G2S2) & Group III included 20 patients (G3S3), where G=The degree of necro-inflammatory process & S=Stage of liver fibrosis. All patients were investigated for the presence of: cryo-globulin, anti neutrophil cytoplasmic (ANCA), anti-liver kidney microsomes (LKM), anti-double stranded DNA, (ds-DNA), anti-nuclear (ANA), anti-mitochondrial (AMA) and anti smooth muscle (ASMA) auto-antibodies. The following results were obtained: ANCA, LKM, ds-DNA, ANA, ASMA, AMA and cryoglobulin were detected in 83/109 (76.1%), 32/109 (29.4%), 23/109 (21.1%), 38/109 (34.9%), 25/109 (22.9%), 5/109 (4.6%) and 60/109 (55%) of chronic HCV respectively. A highly significant positive correlation was found only between ANCA auto-antibodies and cryoglobulin versus grades of liver cirrhosis. Using ANCA, cryoglobulin, age and gender as covariates and by logistic regression analysis, Odds ratio (OR) revealed that these covariates were significant predictors of cirrhosis that add significance to the model according to the sequence: ANCA, cryoglobulin, age and gender suggesting that these covariates associate significantly with development of cirrhosis in HCV patients and that they are significant predictors of liver cirrhosis in HCV patients. The high prevalence of autoantibodies in chronic HCV patients suggests that HCV may trigger an autoimmune reaction, but most probably do not indicate a distinct autoimmune mechanism. Cryoglobulins and ANCA may be a useful prognostic indicator for increased risk of cirrhosis in chronic HCV patients. Follow up studies are recommended. PMID- 15124752 TI - HCV and associated concomitant infections at Sharkia Governorate, Egypt. AB - The natural history of hepatitis C virus (HCV) infection has a highly variable course. Many patients develop chronic infection, with its consequent risk of cirrhosis, liver failure and hepatocellular carcinoma. A key question is whether patients at high risk of disease progression can be distinguished from those with relatively benign disease course. The disease progression is influenced by other factors such as duration of infection, age at infection, sex, co-infection with hepatitis B virus (HBV), Epstein Bar virus (EBV), cytomegalovirus (CMV), the level of HCV viraemia and its type. Other endemic infections in the community as bilharziasis may have a role in progression of the condition to serious complications. These factors are correlated with newly proposed grades and stages of the disease. The studied (109) cases were divided into 6 groups according to the concomitant infection with HCV. The result proved that groups 1, 3 & 5 had a higher level of viraemia than other groups, and to be the high-risk groups as 56.4% and 34.6% were in G2S2 and G3S3, respectively. All cases of liver cell dysplasia and hepatocellular carcinoma in this study were seen in these groups. The conclusion showed that these factors play an important role in the progression of HCV infection. Death of the patients of this progressive condition occurs in younger age and is more due to liver failure than to HCC. PMID- 15124753 TI - Non-invasive markers and predictors of severity of hepatic fibrosis in HCV patients at Sharkia Governorate, Egypt. AB - Liver biopsy is thought mandatory for management in patients with hepatitis C virus infection (HCV) especially for histopathological grading and staging of the disease to assess suitability for treatment and monitoring disease progression. However, tracking of liver disease progression can't rely on repeated biopsies. The study aimed to evaluate two significant items, we try to develop and validate a non-invasive predictive tool to assess hepatic necro-inflammation and fibrosis. Also, to determine factors that associate severity of hepatic pathology in HCV infected Egyptian patients particularly at Sharkia G. The study included 109 patients with detectable HCV by Real Time-PCR. The patients were classified into three different pathological stages and grades according to the new concept of histopathoglical staging and grading. The different clinical, biochemical, virological and ultra-sonographic parameters were assessed and analyzed and the variables that showed significant association with histopathological staging and grading were included in multivariate logistic regression analysis. The regression model revealed that, platelet count, matrix metalloproteinase-9 (MMP 9), portal vein diameter, splenic longitudinal axis, alanine transaminase, aspartate transaminase and viral load were the factors that add significance to the model in decreasing order of significance. From these findings we generate a new score ranged from 0-9. The score model was applied to our patients to assess its validity where it proved to be accurate in discriminating patients with mild inflammation and fibrosis (sensitivity 81.8%, specificity 80.5% and accuracy 80.7%) and more accurate in detecting patients with cirrhosis (specificity 96.6%, sensitivity 80% & accuracy 93.6%) but less accurate in detecting patients with moderate to severe fibrosis (specificity 66.7%, sensitivity 68.7% & accuracy 67.9%). Also the results revealed that, co-infection with schistosomiasis, old age > or = 45 years and positive history of blood transfusion as a source of infection was significantly associated with severe hepatic pathology. It is concluded that, the score model can't completely replace liver biopsy but at least it could be used to substantially reduce the number of liver biopsies done in patients with HCV infection in assessing disease progression during follow up. Also, it can be used to make decisions about treatment in patients who have contraindications to or who refused liver biopsy. Co-infection with schistosomiasis, age > or = 45 and positive history of blood transfusion in patients with HCV warrant special attention with more intensive follow up. These factors may play a major role in forecasting the course of HCV as well as in determining the therapeutic approach in each case. PMID- 15124754 TI - The role of aflatoxin-contaminated food materials and HCV in developing hepatocellular carcinoma in Al-Sharkia Governorate, Egypt. AB - Aflatoxins, particularly aflatoxin B1 (AFB1) have been recognized as one of the most potent chemical carcinogen. In Egypt, HCV is prevalent. The progressive nature of HCV-related liver diseases was found to be influenced by other factors. In this paper, the role of aflatoxin contamination in the onset of liver cancer in HCV-infected patients was studied. The quantitative identification of the possible aflatoxins contamination in six urban and eleven rural areas using high performance liquid chromatography technique, revealed that corn, wheat, pea nut, lupine "termis", white rice, cowpea "lobiya", fava bean and brown rice showed the prevalence of AFB1 to be 64.7%, 53%, 53%, 47%, 47%, 41%, 29.4% & 29.4% respectively. A positive correlation was found between aflatoxin and positive HCV PCR together with liver disease progression to G3S3, the indicative of hepatocellular carcinoma. Such correlation was not fully understood, but the oncogene amplification caused by HCV-infection may be aggravated by the consumption of aflatoxin contaminated raw food materials or their products. PMID- 15124755 TI - Interferon antibodies: do they alter the beneficial effects of beta-interferon upon relapse rate in multiple sclerosis? PMID- 15124756 TI - Longitudinal analyses of the effects of neutralizing antibodies on interferon beta-1b in relapsing-remitting multiple sclerosis. AB - We have analysed data on exacerbation rates, Expanded Disability Status Scale (EDSS) scores, and lesion burdens using the results of two neutralizing antibody (NAB) assays (CPE and MxA) from the pivotal relapsing-remitting multiple sclerosis (MS) trial of interferon beta-1b (IFNB) with a longitudinal approach, where the influence of NABs in individual patients is assessed by comparing responses during NAB-positive and NAB-negative periods. There are apparent influences on exacerbation rate related to dose of IFNB, titer level, and duration of positivity. With the MxA assay, exacerbation rates after switching to NAB-positive status are estimated to be 28% higher [95% confidence interval (CI): (-15%, 92%)] and -2% higher [95% CI: (-21%, 21%)] on the low- and high-dose IFNB arms, respectively. When compared with all NAB-negative periods, exacerbation rates during NAB-positive periods are estimated to be 29% higher [95% CI: (0%, 67%)] and 18% higher [95% CI: (0%, 40%)] on the low- and high-dose IFNB arms, respectively. When NAB-positive patients again become NAB-negative, no evidence of increased exacerbation rates could then be demonstrated. More detailed exploratory analyses indicate that the effects are most evident in the approximately 20% of patients developing high titers. In these patients, the influence of NABs may be self-limited, as titers often diminish or NABs become undetectable with time. PMID- 15124757 TI - Interferon beta-1a in relapsing multiple sclerosis: four-year extension of the European IFNbeta-1a Dose-Comparison Study. AB - BACKGROUND: Multiple sclerosis (MS) is a chronic disease requiring long-term monitoring of treatment. OBJECTIVE: To assess the four-year clinical efficacy of intramuscular (IM) IFNbeta-1a in patients with relapsing MS from the European IFNbeta-1a Dose-Comparison Study. METHODS: Patients who completed 36 months of treatment (Part 1) of the European IFNbeta-1a Dose-Comparison Study were given the option to continue double-blind treatment with IFNbeta-1a 30 mcg or 60 mcg IM once weekly (Part 2). Analyses of 48-month data were performed on sustained disability progression, relapses, and neutralizing antibody (NAb) formation. RESULTS: Of 608/802 subjects who completed 36 months of treatment, 493 subjects continued treatment and 446 completed 48 months of treatment and follow-up. IFNbeta-1a 30 mcg and 60 mcg IM once weekly were equally effective for up to 48 months. There were no significant differences between doses over 48 months on any of the clinical endpoints, including rate of disability progression, cumulative percentage of patients who progressed (48% and 43%, respectively), and annual relapse rates; relapses tended to decrease over 48 months. The incidence of patients who were positive for NAbs at any time during the study was low in both treatment groups. CONCLUSION: Compared with 60-mcg IM IFNbeta-1a once weekly, a dose of 30 mcg IM IFNbeta-1a once weekly maintains the same clinical efficacy over four years. PMID- 15124758 TI - Multiple sclerosis attacks are associated with picornavirus infections. AB - Multiple sclerosis (MS) is a chronic demyelinating disease of the central nervous system, which often follows a relapsing-remitting (RR) course with discrete attacks. MS attacks have been associated with upper respiratory infections (URIs), but the specific viruses responsible have not been identified. We studied a cohort of 16 RRMS patients experiencing URI and followed them for clinically identifiable attacks. The viral causes of 21 separate URIs were investigated using culture and polymerase chain reaction (PCR) of nasal swab specimens, and by serology. Sibleys 'at-risk' period for MS attacks, beginning two weeks before and continuing for five weeks after a URI, was used for the analysis. Seven of the nine (78%) URIs due to picornaviruses were associated with an MS attack during the at-risk period. By contrast, only two of 12 (17%) picornavirus-negative URIs were associated with an MS attack (P = 0.01). The possible role of picornaviruses in the pathogenesis of MS deserves further study. PMID- 15124759 TI - Influence of CCR5 delta32 polymorphism on multiple sclerosis susceptibility and disease course. AB - The CCR5 chemokine receptor has been implicated in the pathogenesis of multiple sclerosis (MS). We carried out an allelic association study using a deletion polymorphism in the coding region of the CCR5 gene in 331 relapsing-remitting (RR) and secondary progressive (SP) MS patients, 108 primary progressive (PP) MS patients and 230 healthy controls. Of the 331 RR and SPMS patients, 172 were recruited from specialist clinics and 159 from a population survey. Disease severity was assessed clinically using the Expanded Disability Status Scale (EDSS) and used to calculate a progression index for each patient (defined as EDSS divided by duration of disease). No significant difference in distribution of the CCR5 delta32 allele was observed between the 331 RR/SPMS patients and controls, between the 108 PPMS patients and controls or between the PPMS and RR/SPMS groups. Furthermore, no differences in rate of disease progression were detected between carriers and noncarriers of the delta32 allele. In the population-based group of RR/SPMS patients, carriage of the CCR5 delta32 polymorphism was associated with a lower age at disease onset (mean age 26.562 versus 31.065 years, P = 0.003). However, no significant differences in age of onset were present in the PPMS group or in a second RRMS population. These results suggest that the CCR5 delta32 polymorphism is not a major determinant of susceptibility to develop MS in the population under study, and conflict with a previously reported association between CCR5 delta32 carriage and a better prognosis. PMID- 15124760 TI - Genotypes at the APOE and SCA2 loci do not predict the course of multiple sclerosis in patients of Portuguese origin. AB - Multiple sclerosis (MS) is a demyelinating disease that affects about one in 500 young Europeans. In order to test the previously proposed influence of the APOE and SCA2 loci on susceptibility to MS, we studied these loci in 243 Portuguese patients and 192 healthy controls and both parents of 92 patients. We did not detect any significant difference when APOE and SCA2 allele frequencies of cases and controls were compared, or when we compared cases with different forms of the disease. Disequilibrium of transmission was tested for both loci in the 92 trios, and we did not observe segregation distortion. To test the influence of the APOE epsilon4 and SCA2 22 CAGs alleles on severity of disease, we compared age at onset and progression rate between groups with and without those alleles. We did not observe an association of the epsilon4 or the 22 CAGs alleles with rate of progression in our total patient population; allele epsilon4 was associated with increased rate of progression of MS in a subset of patients with less than 10 years of the disease. However, globally in the Portuguese population, the APOE and SCA2 genes do not seem to be useful in the clinical context as prognostic markers of this disorder. PMID- 15124761 TI - Win 55212-2, a cannabinoid receptor agonist, attenuates leukocyte/endothelial interactions in an experimental autoimmune encephalomyelitis model. AB - Multiple sclerosis (MS) is the most common of the immune demyelinating disorders of the central nervous system (CNS). Leukocyte/endothelial interactions are important steps in the progression of the disease and substances that interfere with these activities have been evaluated as potential therapeutic agents. Cannabinoid receptor agonists have been shown to downregulate immune responses and there is preliminary evidence that they may slow the progress of MS. The purpose of this investigation was to determine how cannabinoid receptor agonists interfere with leukocyte rolling and adhesion. This was investigated in an experimental autoimmune encephalomyelitis (EAE) model using six to eight week old C57BL/6 mice. Mouse myelin oligodendrocyte protein and pertussis toxin were used to induce EAE. WIN 55212-2, CB1 and CB2 antagonist were given. By use of in vivo intravital microscopy, leukocyte/endothelial interactions were evaluated via a cranial window implanted two days before. The results demonstrated that EAE increases leukocyte rolling and firm adhesion in the brain, and that this increased leukocyte/endothelial interaction can be attenuated by administration of WIN 55212-2. Furthermore, use of the selective antagonists for the CB1 receptor (SR 141716A) and the CB2 receptor (SR144528) in this study demonstrated that the cannabinoid's inhibitory effects on leukocyte/endothelial interactions can be mediated by activating CB2 receptor. PMID- 15124762 TI - Cytokine mRNA expression in patients with multiple sclerosis and fatigue. AB - BACKGROUND: Fatigue is one of the most common disabling symptoms in patients with multiple sclerosis (MS), but the putative role of proinflammatory cytokines remains to be elucidated. METHODS: Thirty-seven patients (27 women, 10 men) with relapsing remitting (n = 29) and secondary progressive (n = 8) MS, aged 41.0 +/- 10.2 years, were studied. Fatigue was assessed by Krupp's Fatigue Severity Scale (FSS). Cytokine mRNA expression for interferon (IFN)-gamma tumor necrosis factor (TNF)-alpha and interleukin (IL)-10 were measured by real time RT PCR. Autonomic function was evaluated by standard tests for parasympathetic and sympathetic function, as well as by serum levels of norepinephrine and epinephrine. RESULTS: Median levels of TNF-alpha mRNA expression were significantly higher in MS patients with (FSS > or = 4.0 and > or = 5.0, n = 26 and n = 14, respectively) than in those without fatigue (FSS < 4.0, n = 11). No differences were seen for IFN-gamma and IL-10 mRNA expression. Cytokine levels were not correlated to autonomic tests or to serum catecholamine levels. CONCLUSIONS: These results suggest that TNF-alpha, as a principal proinflammatory mediator, is associated with MS-related fatigue. This is in support of a pathogenic role of the MS related inflammatory process in the development of fatigue. PMID- 15124763 TI - Risk of bone loss in men with multiple sclerosis. AB - CONTEXT: Osteoporosis and the increased fracture risk associated with osteoporosis become apparent in men approximately 10 years later than women. However, in recent studies, approximately 20% of healthy men in the age range 55 64 years were found to be osteopenic. Emerging data suggest a significantly increased prevalence of osteoporosis in men and women with multiple sclerosis (MS) compared to age-matched controls, but no specific clinical testing recommendations are available for men. OBJECTIVE: To determine the proportion of male MS patients with osteoporosis and to identify the factors associated with the reduction in bone mass. DESIGN: Consecutive male MS patients seen at our MS clinic were screened with dual-X-ray absorptiometry (DEXA) scan for determining the bone mineral density (BMD). All patients had neurological Expanded Disability Status Scale (EDSS) evaluation. The results were compared to healthy age-matched male reference population using the Z score and to a cohort of women MS patients and women controls. Calcium, total testosterone, sex-hormone binding globulin (SHBG), 25-hydroxy-vitamin-D, and parathyroid hormone (PTH) were evaluated in male patients with decreased BMD. Relevant data on body mass index (BMI), medication, alcohol consumption, smoking, and sexual dysfunction were recorded. SETTING: Academic MS Centre. PATIENTS AND OTHER PARTICIPANTS: Forty consecutive male MS patients, age mean 51.2 +/- 8.7 years, and mean EDSS of 5.8 +/- 1.9 were evaluated with DEXA scan. Of these, 17.5% patients were relapsing-remitting (RR) MS, 57.5% were secondary progressive (SP) MS and 25% were primary progressive (PP) MS. MAIN OUTCOME MEASURE: Proportion of male MS patients with reduced BMD at the lumbar spine and femoral neck. RESULTS: Thirty-two (80%) of our patients had a reduced bone mass of either lumbar spine or the femoral neck; of these 17 patients (42.5%) had osteopenia and 15 patients (37.5%) had osteoporosis. Twenty one per cent (eight out of 38 patients) had vertebral, rib or extremities fractures. Multivariate linear regression analysis indicated that the EDSS (P < 0.0001) and BMI (P = 0.0004) were the important factors associated with low BMD at the femoral neck and the EDSS was the important factor (P = 0.0017) associated with low BMD at the lumbar spine. The same factors emerged as significantly associated with the corresponding Z scores, which are corrected for age and sex. No clear association between intravenous steroid therapy and BMD was evident in the multivariate analysis. Low levels of 25-hydroxy-vitamin-D were seen in 37.5% of patients. CONCLUSIONS: The proportion of male MS patients with reduced bone mass is high and disproportionate to their age and ambulation, consistent with an association between the MS disease process and pathological bone loss. Increased awareness and bone density screening of male and female MS patients over 40 years of age is warranted. PMID- 15124764 TI - Bcl-2 and its homologues in the brain of patients with multiple sclerosis. AB - We measured the mRNA expression levels of molecules involved in scavenging free radicals and in apoptosis within normal appearing white and gray matter (NAWM and NAGM, respectively) and chronic active plaque containing frontal lobe specimens of patients with multiple sclerosis (MS). While no regional differences were detected in the mRNA levels of free radical scavengers, Bcl-XL was higher in plaques than in NAWMs, and both Bak and Bcl-2 were found to be increased in correlation with an immune marker (beta2-microglobulin--beta2Mg) in NAWM and plaque compared with corresponding cortical regions. We did not measure a similar white-gray matter difference in the expression of the latter genes in brains of normal or Alzheimer disease controls. This finding indicates that both pro- and anti-apoptotic mechanisms are activated, not only within but also outside of plaques. PMID- 15124765 TI - Progressive change in primary progressive multiple sclerosis normal-appearing white matter: a serial diffusion magnetic resonance imaging study. AB - In spite of marked disability, patients with primary progressive multiple sclerosis (PPMS) display smaller lesion volumes on conventional magnetic resonance imaging (MRI) compared with other forms of multiple sclerosis (MS). Hence, damage to the normal-appearing brain tissue (NABT) may play an important role in explaining the pathogenesis of disability in PPMS. Diffusion-weighted MRI (DW-MRI) probes water diffusion in vivo that can be altered by pathologic changes. Using DW-MRI we investigated diffusion in the NABT of 15 patients with PPMS over one year. The average apparent diffusion coefficient (ADCav) was measured in 10 regions of interest located in the normal-appearing thalamus and the normal-appearing white matter (NAWM). Six healthy subjects served as a reference. In contrast to healthy subjects, patients with PPMS showed an increment within 12 months of the ADCav in NAWM which was associated with an increase of the T2- and T1-lesion volumes. The ADCav in frontal NAWM was associated with disability as measured by the MS Functional Composite Measure. Serial DW-MRI depicts progressive changes in the NAWM of patients with PPMS. Our preliminary findings suggest that the processes causing structural damage in NAWM and lesions in patients with PPMS are partially linked and that changes of water diffusion in NAWM depicted by DW-MRI are clinically relevant. PMID- 15124766 TI - Diffusion tensor imaging in multiple sclerosis: a tool for monitoring changes in normal-appearing white matter. AB - Our objectives were to determine the reproducibility of diffusion tensor imaging (DTI) in volunteers and to evaluate the ability of the method to monitor longitudinal changes occurring in the normal-appearing white matter (NAWM) of patients with multiple sclerosis (MS). DTI was performed three-monthly for one year in seven MS patients: three relapsing-remitting (RRMS), three secondary progressive (SPMS) and one relapsing SP. They were selected with a limited cerebral lesion load. Seven age- and sex-matched controls also underwent monthly examinations for three months. Diffusivity and anisotropy were quantified over the segmented whole supratentorial white matter, with the indices of trace (Tr) and fractional anisotropy (FA). Results obtained in volunteers show the reproducibility of the method. Patients had higher trace and lower anisotropy than matched controls (P < 0.0001). Over the follow-up, both Tr and FA indicated a recovery after the acute phase in RRMS and a progressive shift towards abnormal values in SPMS. Although this result is not statistically significant, it suggests that DTI is sensitive to microscopic changes occurring in tissue of normal appearance in conventional images and could be useful for monitoring the course of the disease, even though it was unable to clearly distinguish between the various physiopathological processes involved. PMID- 15124767 TI - Multiple sclerosis and alcohol: a study of problem drinking. AB - Multiple sclerosis (MS) patients are known to be at increased risk from mood disorders and suicidal ideation. Although these are often associated with alcohol disorders, the drinking habits of MS patients have not been well studied to date. Our study assessed drinking patterns in 140 MS patients, focusing on a possible link between problem drinking and mood and anxiety disorders. Lifetime psychiatric diagnoses were ascertained using the Structured Clinical Interview for DSM-IV disorders (SCID-IV). Results demonstrate that one in six MS patients drink to excess over the course of their lifetime. Those with a history of problem drinking display a higher lifetime prevalence of anxiety (P = 0.006), but not mood disorders. There were also significant associations between problem drinking and a lifetime prevalence of suicidal ideation (P = 0.006), substance abuse (P = 0.001), and a family history of mental illness (P = 0.008). Clinicians should be aware of the possibility of problem drinking in MS patients, and how this may complicate the course of their disease. Clues to problem drinking in MS patients are the presence of a positive family history of mental illness and prominent anxiety. PMID- 15124768 TI - What affects your MS? Responses to an anonymous, Internet-based epidemiological survey. AB - Evolving information technology has raised the possibility of new methods of data collection in multiple sclerosis (MS) research. An anonymous, self-report, Internet-based survey was developed, which asked people with MS their opinion on how various extrinsic factors affected their condition. From September 2001 to July 2002, a total of 2529 people completed the questionnaire. The demographic and clinical profiles of the anonymous respondents indicated that most were likely to have MS. Common factors reported as beneficial were cannabis, cold baths, meditation and dietary factors. Common adverse factors reported were high stress, exposure to high temperatures and viral infections. There was an increasing report of high temperatures as being adverse with increasing respondent age (test for trend, P < 0.001). The adverse report of high temperatures correlated significantly with the report of strong sunlight apparently making MS worse (r = 0.35, P < 0.0001). In Australia, high temperatures were more likely to be reported as adverse in warmer, lower latitude regions. The association between strong sunlight as adverse and age or region did not persist after adjustment for high temperatures. Thus, this apparent adverse factor appeared to relate to solar heat, not solar light. People with MS may risk vitamin D deficiency because of sun avoidance due to heat-related fatigue or intolerance. This is of clinical significance not only for bone health but because vitamin D may have beneficial immunomodulatory properties. The present study provides new information from people with MS on factors that may influence symptoms or clinical course. This information will now be used in the design of formal epidemiological cohort studies. PMID- 15124769 TI - Exercise capacity, disability and leisure physical activity of subjects with multiple sclerosis. AB - The purpose of the present study was to examine exercise capacity and its relationship to neurological disability as measured using the Expanded Disability Status Scale (EDSS) and to leisure physical activity in subjects with multiple sclerosis (MS). Thirty-four men and 61 women (mean age 44 +/- 6.7 years, mean disease duration 5.7 +/- 6.4 years) with mild to moderate disability (EDSS range 1.0-5.5) participated. They underwent an incremental exercise test on a leg cycling ergometer. Leisure physical activity was measured using a questionnaire. Peak oxygen uptake (VO2peak) in men was 27.0 +/- 5.2 mL/kg/min, and in women 21.7 +/- 5.5 mL/kg/min. The disability correlated inversely with the VO2peak both in men (r = - 0.50, P = 0.004) and in women (r = - 0.25, P = 0.05). No correlation between disease duration and VO2peak was found. In a multivariate regression analysis, neurological disability was confirmed as a predictor of VO2peak. No evidence of a relationship between leisure physical activity and VO2peak was found. A main finding was that disability and exercise capacity are inter related, even in subjects who are not severely handicapped (84% had an EDSS of < 4.0). The level of disability should be taken into account in the planning of aerobic exercise programs for fully ambulatory MS subjects. PMID- 15124770 TI - Caregivers of people with multiple sclerosis: experiences of support. AB - The aim of this phenomenological study was to gain an understanding of the experiences of a group of caregivers of people with multiple sclerosis (MS). Sixteen caregivers from Northern Ireland and the Republic of Ireland participated in focus group interviews. The theme of support, either sought or received, emerged as a major aspect of the experiences described. Caregivers' feelings about, and experiences of, support appeared to change over time. Four common phases that caregivers experienced in relation to support were identified as: 'rejecting', 'resisting, 'seeking' and 'accepting' support. This paper will present and discuss these four phases. The study findings highlight the complexity of issues surrounding a caregiver's decision to seek and accept support. It is hoped that the phases identified within this study are useful in depicting how caregivers of people with MS may progress through stages in their desire for, and acceptance of support. Findings from this study are useful to healthcare professionals who work with people with MS and their caregivers by increasing awareness that a caregiver's attitude toward and acceptance of support changes over time. PMID- 15124771 TI - SaGAS, the Short and Graphic Ability Score: an alternative scoring method for the motor components of the Multiple Sclerosis Functional Composite. AB - The timed performances of the 10-m timed walk (TMTW) and the nine-hole peg test (NHPT) of 881 consecutive patients with multiple sclerosis (MS) undergoing a rehabilitation stay, were expressed as a logarithmic function of time in two subscores to form a composite score called the Short and Graphic Ability Score (SaGAS). The subscores (sS) were constructed in such a way that any interval of 0.5 unit corresponds to a change of 25% in the tests. The SaGAS was computed as the mean of four subscores: SaGAS = (2 x TMTWsS + NHPTsS right hand + NHPTsS left hand). With the aid of a nomogram, the timed values of the tests are easily transformed into the corresponding subscores, which are then displayed graphically to facilitate follow-up over time. The correlation coefficients between the SaGAS and the two motor components of the MS Functional Composite (MSFC) (r = 0.987), the Expanded Disability Status Scale (EDSS)(r = -0.83), the Nottingham EADL Index (r = 0.80) and the Rivermead Mobility Index (RMI) (r = 0.90) were all statistically significant (P < 0.001), supporting the validity of the measure. SaGAS had a similar sensitivity to the RMI, but was significantly more sensitive than the EDSS in detecting changes occurring during the rehabilitation stay (14.9% versus 5.0%; P < 0.001) and over a one-year follow-up (35.3% versus 19.7%; P < 0.001). Compared with the motor components of the MSFC, with which it shares several features, SaGAS has several advantages: it does not depend on the stratification of the study population; it does not skew the results of the NHPT towards improvement at the lower end; and it offers an independent assessment of both hands. SaGAS is a simple, intuitive, nonphysician based measure, which could provide consistent scoring in future clinical trials. PMID- 15124772 TI - C1-inhibitor activity and plasma concentrations in patients with monoclonal gammopathies: implications for treatment with recombinant interferon-beta 1b. PMID- 15124773 TI - The humoral and cellular immune response of sheep against Borna disease virus in endemic and non-endemic areas. AB - Borna Disease (BD) is a mostly fatal disease of horses and sheep endemic in central Europe. Antibodies to Borna disease virus (BDV) have been described in sheep and other species living in BD non-endemic areas. Meaningful clinical BDV serology is hampered by difficulties in defining serological cut-offs, which require the investigation of populations from endemic areas. Here we studied BD serology in sheep from endemic and non-endemic areas of similar geography in Switzerland. Antibodies to BDV antigens were detected by ELISA and indirect immunofluorescence analysis (IFA) only in sera from 3 of 6 sheep with autopsy confirmed BD. One serum was positive by IFA but not by ELISA, while 2 sera were negative in both assays, indicating that not all diseased animals develop BDV specific antibodies. Six % of clinically healthy animals (6/106) from an endemic area and 2% from a non-endemic area (4/192) had serum antibody to either BDV p40 or p24 as detected by ELISA. None of the animals showed a cellular immune response to BDV p40. In some healthy sheep from the endemic area, serum antibody titers to BDV p24 antigen remained elevated over several months without onset of disease symptoms. Infections with either BDV or related viruses may thus occur at low frequency in sheep from non-endemic areas leading to the production of antibodies to BDV antigens. We further propose viral strain differences or environmental factor(s) may determine the clinical outcome. PMID- 15124774 TI - [Sonographic examinations of the intestines in calves]. AB - The aim of this investigation was to describe the normal ultrasonic appearance of the intestinal loops of calves at different stages of their adolescence and at various intervals after suckling and to determine reliable reference values. Therefore the abdomen of 20 clinically healthy calves have been examined with a 5 MHz sector probe on the right and left flank of each animal and the exact position, the largest diameter, the thickness of the walls, the peristaltic activity and the appearance of the contents of each part of the intestines described. Most often, the pars cranialis duodeni could be seen ventral of the right costal arch as well as in the right flank, with younger calves it also showed in the left flank sometimes. The position could be changed considerably depending on the period elapsed since the last suckling time. The duodenum descendens and the duodenum ascendens could be traced in the right flank. The thickness of the wall was found to measure between 2 and 3 mm throughout the whole duodenum. The examination of the jejunum and the ileum was basically done in the right flank. In cases where the rumen was not yet fully developed visualization was also possible in the left flank. Most often these parts were seen in cross-section displaying permanent peristaltic activity. With the younger calves the large intestine could equally be traced in both flanks. In older animals it could be recognized as voluminous hollow organ filled with gas, or in case of the colon ascendens as garlandshape. In the jejunum and ileum as well as in the large intestine the thickness of the wall measured between 1 and 2 mm. Each part of the intestine showed an increase in diameter as the calves grew older. The peristaltic activity increased during two hours after suckling and during this time the echoing level of the contents was lower. PMID- 15124775 TI - Endogenous TSH in the diagnosis of hypothyroidism in dogs. AB - To determine whether measurement of canine thyrotropin (cTSH) would aid in the diagnosis of hypothyroidism, serum samples of 65 dogs with clinical signs suggestive of hypothyroidism were evaluated. Diagnosis was confirmed in 26 dogs and excluded in 39 dogs based on TSH-stimulation testing. Total thyroxine (T4) was significantly lower and cTSH significantly higher in hypothyroid dogs compared to euthyroid dogs. Canine TSH was above (> 0.6 ng/ml) in 15 (57.7%) and below the upper limit of the reference range in 11 (42.3%) of the hypothyroid dogs. All of the euthyroid dogs had a cTSH < 0.6 ng/ml. In all dogs with a cTSH above the upper limit of the reference range hypothyroidism could be confirmed. Therefore, our results show that measurement of cTSH has an excellent specificity (100%) and is a valuable tool in confirming canine hypothyroidism. However, due to the low sensitivity of cTSH assays (60%), it can not be recommended to exclude the disease. PMID- 15124776 TI - Oral health and its determinants among Mongolian dentists. AB - This study compares the dental health of dentists with that of their population counterparts, and relates dentists' oral health to oral health behavior and professional preventive knowledge and orientation. A questionnaire requesting information on the dentists' oral health, preventive knowledge, and orientation and oral health behavior, was targeted at all actively practicing dentists (n = 250) in the Mongolian capital of Ulaanbaatar. A self-reported dentigram served as the basis for assessing dental health, while the 1996 National Oral Health Survey provided data for comparison with population counterparts. The response rate was 98%. Compared to their general-population counterparts, younger dentists exhibited a similar level of caries experience (DMFT) and missing teeth (NMT), whereas older dentists exhibited a lower level. In all age groups, mean numbers of decayed teeth (DT) among dentists were much smaller than for their population counterparts, whereas mean numbers of filled teeth (FT) were much higher. Of all, 81% reported brushing their teeth twice daily or more, 62% used fluoride containing toothpaste always or almost always, and 52% consumed sugar-containing foods between main meals less than once a day. The better oral health behavior of dentists was best explained by their greater preventive knowledge and their better dental health by better oral health behavior in logistic models. It is concluded that among these dentists there is room for improvement of their own oral health behavior. Towards this end, specially designed continuing education courses could serve to improve dentists' health-related knowledge and change their oral self-care behavior. PMID- 15124777 TI - Progression of radiographic changes in the temporomandibular joints of patients with rheumatoid arthritis in relation to inflammatory markers and mediators in the blood. AB - The aim of this study was to investigate longitudinal radiographic changes in the temporomandibular joint (TMJ) with clinical involvement of rheumatoid arthritis (RA) and its relation to the blood level of inflammatory mediators and markers. Sixteen patients were investigated by computed tomography on two occasions 25-46 months apart. The radiographs were assessed independently for changes in presence of erosions, sclerosis, flattening, osteophytes, and subchondral pseudocysts. The serum (S) or plasma (P) concentrations of C-reactive protein (CRP), thrombocyte particle concentration, scrotonin (S-5-HT and P-5-HT), tumor necrosis factor alpha, interleukin-1 receptor antagonist, tumor necrosis factor soluble receptor type II, interleukin-1 soluble receptor type II (P-IL-1sRII) and interleukin 6 as well as the erythrocyte sedimentation rate (ESR) were measured. The radiographic status showed no consistent or significant change during the observation period, but the individual variation was considerable. The radiographic signs of erosion and sclerosis varied most. Regression of erosions was associated with high S-5-HT and P-IL-1sRII, while progression of erosions was associated with high P-5-HT. Regression of sclerosis was associated with an increase in P-5-HT and high ESR. Progression of flattening was associated with high CRP. In conclusion, this study indicates that the progression of radiographic changes that occurs in the TMJ of patients with well-controlled RA during a period of 25-46 months seems to be related to the blood levels of CRP, 5-HT, and IL-1sRII. However, only minor progression can be expected to occur, and with considerable individual variation. PMID- 15124779 TI - Beliefs about professional ethics, dentist-patient communication, control and trust among fearful dental patients: the factor structure of the revised Dental Beliefs Survey. AB - The revised version of the Dental Beliefs Survey (DBS-R), intended to measure three dimensions of the patient dentist relationship as perceived by the patient, namely Ethics, Communication and Control, was tested in a confirmatory factor analysis. Five different models for the internal structure of the questionnaire were tested. The final model, a 5-factor solution, basically including the 3 assumed dimensions as well as the re-introduced dimension Trust covered in the first version of the DBS plus a global factor including all items, yielded an acceptable fit. This model also omits 4 items from DBS-R, i.e. items 3, 11, 18, and 28. The reduced DBS-R is recommended for clinical use when assessing the patient's perception of the relationship to the dentist, including both trust and ethical behavior. PMID- 15124778 TI - Mineralization of dentin induced by treatment with bioactive glass S53P4 in vitro. AB - Dentin hypersensitivity can be managed to occlude dentin tubules, but none of the agents used are components of natural dentin. Using a calcium phosphate precipitation (CPP) method, dentin tubules can be occluded with a calcium phosphate (CaP) layer similar to the major inorganic component of dentin. The CPP method utilizes acidic pH conditions, such as etching of dentin, over the course of several dental treatments. A gentler method can be used to produce a CaP layer on the surface of dentin. By treating with bioactive glass S53P4 (BAG), or regular commercial glass (CG), mineralization occurs in physiologically neutral solutions such as simulated body fluid (SBF) and remineralization solution (RMS). After a short period of immersion, silica is dissolved from both types of glass, but the amount of silica released is much greater from BAG than from CG. The dissolved silica is adsorbed on the surface of dentin during the pretreatment procedure and enhances the mineralization of dentin in SBF. After 14 days' mineralization the dentin is fully covered by the CaP layer, but after 14 days' immersion in RMS decalcification of the dentin occurs. Pretreatment with BAG decreases the degree of decalcification of dentin during the mineralization process. These findings suggest that bioactive glass S53P4 can be used as a therapeutic material for mineralization of dentin and its tubules in a physiological environment. PMID- 15124780 TI - A premaxilla with a supernumerary tooth indicating a developmental region with a variety of dental abnormalities: a report of nine cases. AB - The aim of this study was to elucidate whether a premaxilla with a supernumerary tooth has additional dental abnormalities. The study does not include premaxillas with a mesiodens, only premaxillas with parasagittally located supernumerary teeth. Radiographs from eight children, followed from 1 year and 5 months to 11 years and 5 months in the municipal dental health service in Denmark, were analyzed. One patient was seen only once. Dental abnormalities recorded included: invaginations on permanent incisors, resorption of roots of incisors, curved roots of incisors, delayed eruption, and delayed formation of roots. The study shows that there are dental abnormalities within the premaxillary region where the supernumerary tooth is located. This indicates that, apart from the supernumerary tooth, a more widespread deviation from normal conditions including tooth malformation, arrested eruption, and root resorption occur within the premaxillary region. The most important outcome of this study is that in the region with a supernumerary tooth the adjacent incisor shows delayed eruption after surgical removal of the supernumerary tooth. The deviations in root morphology, including root resorption, are not limited to patients who have had orthodontic treatment, but patients who did not receive orthodontic treatment also revealed deviations in root morphology. Therefore the treatment outcome can be different from the expected outcome. PMID- 15124781 TI - Eight-year study on conventional glass ionomer and amalgam restorations in primary teeth. AB - The aim of this randomized clinical study was to compare the longevity and the cariostatic effects of conventional glass ionomer and amalgam restorations in primary teeth placed in everyday practice in the Danish Public Dental Health Service. All restorations inserted during a 7-month period by 14 clinicians in 2 municipalities were included in the study. The sample consisted of 515 conventional glass ionomer restorations and 543 amalgam restorations in 666 children aged between 2.8 and 13.5 years. The restorations were in contact with 592 unrestored surfaces in primary and permanent teeth. The study was terminated after 8 years, with 2% of the restorations in function and 7% patient dropouts. Fifty percent of the teeth restored with glass ionomer and 63% of those with amalgam were exfoliated with the restoration in situ, while 42% of the glass ionomer and 20% of the amalgam restorations had been repaired or replaced. Fracture of restoration, endodontic complication, and loss of retention were the major reasons for failure. The 50% survival time for glass ionomer restorations in all cavity types was 42 months, while the median survival time for amalgam restorations could not be estimated but exceeded 7.8 years (P < 0.001). Progression of caries lesions on tooth surfaces adjacent to amalgam restorations required operative treatment on 30% of the teeth, while only on 16% of teeth adjacent to glass ionomer restorations. The 75% survival time was 40 months for surfaces in contact with glass ionomer compared to 25 months for surfaces in contact with amalgam (P = 0.005). Multivariate analyses were performed in order to assess the influence of a number of factors on the longevity of restorations, occurrence of prevalent failures, and caries treatment of surfaces in contact with the restorations. Owing to the high frequency of failures of the conventional glass ionomer restorations, it was concluded that they are not an appropriate, universal alternative to amalgam for restorations in primary teeth, although they reduce caries progression and the need for operative treatment of adjacent surfaces. PMID- 15124782 TI - A systematic review of Swedish research in orthodontics during the past decade. AB - The aims of this systematic review were to identify the study designs and topics of Swedish orthodontic articles, to elucidate their international position, and to verify in which scientific journals the articles had been published in the past decade. A search of the Medline database for papers published between 1992 and 2002 was made using the Medical Search Heading terms 'orthodontics', 'malocclusion', 'cephalometry', and 'facial bones and growth'. Two independent reviewers selected the articles of Swedish origin and categorized each article according to research design and principal topic. Overall, 15,571 articles in orthodontic research were found, and the Swedish contribution was 1.9% with the majority of these (71.5%) being submitted by universities. Most of the Swedish articles (84.5%) had been published in 10 journals and many high-quality studies with orthodontic interest were published in non-orthodontic journals with higher impact factor scores than the orthodontic journals. Every second study was prospective, and of these, 15 (5.2% of all Swedish articles) were randomized clinical trials (RCTs). It was found that nearly every third study, prospective as well as retrospective, was uncontrolled. The main classification was treatment studies (51.9%), followed by development (18.6%) and diagnostic information (10.7%) studies. Thus, the majority of the articles evaluated therapeutic interventions; however, although the RCT is the preferred study design in evaluation studies, few used this method. In an era focused on evidence-based medicine, studies with an RCT design will be the future challenge for research in the field of orthodontics. PMID- 15124783 TI - Repair bond strength of restorative resin composite applied to fiber-reinforced composite substrate. AB - Delamination or fracture of composite veneers can occur as a result of improper design of the fiber-reinforced composite (FRC) framework. This in vitro study tested the repair bond strength of restorative composite to aged FRC. The substrate was multiphase polymer matrix FRC (everStick) aged by boiling for 8 h and storing at 37 degrees C in water for 6 weeks. The aged substrate surfaces were wet-ground flat with 1200-grit silicon carbide paper and subjected randomly to 5 different surface treatments: 1) An adhesion primer (Composite Activator) and resin (CA), 2) Silane (EspeSil) and resin (SIL-MP), 3) Silane, adhesive primer, and resin (Clearfil Repair) (CF), 4) Air particle-abrading (CoJet), silane, and resin (CJ-SIL-MP), 5) Resin (Scotchbond Multipurpose Resin) only as control (MP). Restorative composite resin (Z250) was added to the substrate in 2 mm layer increments and light-cured. Subsequently, every surface treatment group was divided into 2 subgroups of 12 specimens each. The specimens were either 48 h water-stored or thermocycled (6000 x 5-55 degrees C). The shear bond strengths of composite resin to FRC were measured at a crosshead speed of 1.0 mm/min. The data were analyzed by ANOVA for factors 'treatment type' and 'storage condition'; Tukey's post-hoc tests and Weibull analysis were performed. ANOVA showed a significant difference as a function of surface treatment (P<0.05) and storage condition (P<0.05). The CJ-SIL-MP group showed highest bond strength and Weibull modulus after thermocycling. Repair of multiphase polymer matrix FRC may show reliable bond strength when silane treatment is used along with air-particle abrading. PMID- 15124784 TI - Secondary caries detection by DIAGNOdent and radiography: a comparative in vitro study. AB - DIAGNOdent is now well documented for detection of primary occlusal caries, but not as yet for secondary caries. The aim of this study was to investigate the application of DIAGNOdent for in vitro detection of secondary caries. The material comprised 66 extracted teeth: 48 with amalgam restorations and 18 with tooth-colored restorations. Digital images of occlusal surfaces were captured for each specimen. The teeth were mounted in groups of 3 or 4 in plaster blocks simulating their anatomical positions. Bitewing radiographs of each block were assessed for secondary caries by 5 observers. The margins of each restoration were carefully scanned with DIAGNOdent; the site of the highest reading was localized in digital images; and the corresponding values were registered. Marginal integrity and staining were also documented. The restorations were then removed and the teeth were hemi-sectioned. For verification, two observers working together examined all the cavities under a stereomicroscope at 16 x magnification followed by probing. Sensitivity and specificity for DIAGNOdent and conventional radiography were 0.77/0.81 and 0.65/0.81, respectively. Regarding ROC analyses, the Az values were 0.89 and 0.72 for DIAGNOdent and radiography. For DIAGNOdent, the false-positive fraction included only stained teeth. Cohen's kappa statistics disclosed moderate agreement between the 2 methods, with an agreement of 56%. DIAGNOdent tended to give more positive diagnoses than bitewing radiography. The results indicate that DIAGNOdent may be a helpful tool for detecting secondary caries. PMID- 15124785 TI - Colon injuries: new perspectives. PMID- 15124786 TI - A prospective study of post-traumatic biliary and pancreatic fistuli. The role of expectant management. AB - BACKGROUND: Biliary or pancreatic fistulae occur frequently after operative repair of liver or pancreatic injuries. The existing literature is based on a few retrospective series. The objective of this prospective study is to evaluate the clinical course and efficacy of non-operative treatment of biliary fistuli (BF) and pancreatic fistuli (PF) post-traumatic fistulae. STUDY DESIGN: Patients who following a trauma Laparotomy, developed BF or PF were prospectively followed during a 38-month period (June 1999-August 2001). Demographics, injury type and severity, fistula characteristics and daily output, complications, interventions and cost were recorded. Optimal nutrition, wound management, and control of infection were priorities in fistula management. A fistula was considered completely healed if the patient was on regular diet and the output was zero for at least 48 h. RESULTS: Of 160 patients (injury severity score: 21 + 10) with trauma laparotomy for a liver or pancreatic injury, nineteen patients (12%) developed a fistula (11 BF, 8 PF). No patient died. Infections occurred in 45% (5/11) of BF patients and 50% (4/8) of PF patients. Only two patients required an operation, one for biliopericardium and one more for a pancreatic pseudo-cyst. Fistula management was responsible for more than half of the hospital stay and cost. Patients with PF had longer hospital stays (44 + 28 days) and charges (US dollars 345,000 + 218,000) than patients with BF (22 + 12 days and US dollars 103,000 + 61,000, respectively). CONCLUSIONS: Most post-traumatic BF and PF can be managed non-operatively. BF resolves earlier than PF. Both entities are responsible for substantial increases in hospital length of stay and charges. PMID- 15124787 TI - Gastric rupture from blunt abdominal trauma. AB - Gastric rupture following blunt abdominal trauma is rare, with a reported incidence of 0.02-1.7%. Road traffic accidents remain the most frequent cause. The factors most often implicated in the genesis of this entity are: a history of a recent meal, trauma to the left side of the body and an inappropriate use of seat belts. Splenic injury is generally the most common associated injury. The high morbidity and mortality are directly related to the number of associated injuries, delays in diagnosis and the development of intraabdominal sepsis. We performed a retrospective study of 1300 patients with blunt trauma to the abdomen from 1973 to 2001. Seven patients sustained a gastric rupture (five men and two women). The following associated characteristics were analysed: mechanism of injury, clinical presentation, possible associated injuries and postoperative complications, diagnosis methods and surgical treatment. We found an incidence of gastric rupture of 0.5%. We emphasise an early diagnosis and aggressive surgical treatment as a key to decreasing the mortality and morbidity from this injury. However, in our series, the morbidity is mainly from associated injuries. PMID- 15124788 TI - Effects of molsidomine and lexipafant in hepatic ischaemia--reperfusion injury. AB - PURPOSE: The purpose of this study was to evaluate the effects of nitric oxide donor molsidomine and platelet-activating factor (PAF) antagonist lexipafant on the hepatic IR injury in rats. METHODS: Fifty male Sprague-Dawley rats (200-225 g) were divided into five groups each containing 10 rats; group SO: Sham operation group; group I: hepatic ischaemia group; group IR: ischaemia reperfusion (IR); group M: IR plus pretreatment with molsidomine; group L: IR plus pretreatment with lexipafant. Hepatic ischaemia and reperfusion, each were applied for 45 min. Hepatic specimens were obtained to determine the tissue levels of malondialdehyde (MDA) and histological changes. Blood samples were obtained by cardiac puncture for determination of alanine transaminase (ALT), aspartate transaminase (AST) and lactic dehydrogenase (LDH). RESULTS: The liver damage scores of groups I, IR, M and L were significantly higher than that of group SO (P < 0.001). The liver damage scores of groups IR and M were significantly higher than that of group I (P = 0.009 and 0.0035, respectively). The liver damage scores of groups M and L were significantly lower than that of group IR (P < 0.001 for both M and L). Mean MDA levels of groups I and IR were significantly higher than those of group SO (P < 0.001). Administrations of molsidomine and lexipafant prior to ischaemia-reperfusion (IR) resulted in significant reduction of the MDA values (P < 0.001). A statistically significant (P < 0.001) decrease in the levels of AST, ALT and LDH was observed in groups M and L compared with group IR. CONCLUSION: In conclusion, these observations suggest that pre-treatment with nitric oxide donor molsidomine and PAF antagonist lexipafant before the reperfusion period may be useful in preventing hepatic reperfusion injury. PMID- 15124789 TI - Is early enteral feeding safe in patients who have suffered spinal cord injury? AB - OBJECTIVE: To examine whether enteral feeding is a safe technique to use in the acute stage of spinal cord injury. METHODS: We searched the departmental computerised patient database and clinical records for all patients with spinal cord injuries admitted to the Auckland Hospital Intensive Care Unit (ICU), known as the Department of Critical Care Medicine (DCCM), between January 1988 and December 2000. Patients were included in the study if they had suffered complete spinal cord transection resulting in either paraplegia or quadriplegia. Data was collected for the following variables: length of time to commence enteral feeding, type of enteral feeding, duration of enteral feeding and reasons for interrupting the feed. RESULTS: Thirty-three patients were found and were included in the study. Twenty-seven (82%) of the patients commenced enteral feeding in the DCCM, 25 by nasogastric (NG) and 2 by nasojejunal (NJ) tube. Feeding was commenced a median of 2 days after admission and the median length of enteral feeding was 7.7 days. The main feeding complications that resulted in interrupting the feed were high gastric aspirates. One patient commenced on enteral feeding developed medical complications that prevented continuation. Two patients on NG feeding converted to NJ feeding. CONCLUSION: No major complications associated with enteral feeding were seen in this study. This would indicate that enteral feeding can be safely administered in the acute stage of spinal cord injury provided complications are monitored for daily. PMID- 15124790 TI - Suture characteristics following incubation in synovial fluid or phosphate buffered saline. AB - Selection of an appropriate suture requires a knowledge of many factors. These include its mechanical characteristics, its durability, the period of mechanical effectiveness and the tissue reactivity to the material. Absorbable suture materials were incubated in synovial fluid or phosphate buffered saline, at 37 degrees C, for periods up to 12 weeks. Mechanical testing was performed to determine the yield strength, percentage elongation and stiffness of each material in response to incubation. Significant differences were observed in response to time of incubation, material size ('0' or '2-0'), incubation medium and material properties (P < 0.001). Based on the results obtained Vicryl and PDS appear ideal for short-term and medium term apposition, respectively. Panacryl has more durable mechanical features and may well be suited to long-term tissue apposition, such as tendon repair or arthroplasty. PMID- 15124791 TI - Anthropometrical analysis of cervical spine injuries. AB - To date the principal focus of the mechanism of cervical spine fracture has been directed towards head/neck circumference and vertebral geometric dimensions. However the role of other measurements, including chest circumference and neck length, in a standard cervical fracture population has not yet been studied in detail. Cervical fractures often involve flexion/extension type mechanisms of injury, with the head and cervical spine flexing/extending, using the thorax as an end point of contact. Thus, the thorax may play an important role in neck injuries. STUDY DESIGN: We prospectively studied all patients with cervical spine fractures who were admitted to the National Spinal Injuries Unit from 1 July 2000 to 1 March 2001. Anthropometrical measurement of head circumference, neck circumference, chest circumference, and neck length were analysed. Ages ranged from 18 to 55 years, and all patients with concomitant cervical pathology were excluded from the study. Mechanism of injury involved flexion/extension type injuries in all cases; those with direct axial loading were excluded. A control group of 40 patients (age 18-50 years) involved in high velocity trauma with associated long bone fractures, in whom cervical injury was suspected, but who were without any cervical fracture, or associated pathology, were similarly measured. RESULTS: Our analysis revealed a statistically significant increase in chest size in the male control group versus the male fracture group (97.89 cm versus 94.19 cm, P < 0.05, Student's t-test). There was a correspondingly significant increase in chest circumference between the female controls versus the female fracture group (92.33 cm versus 88.88 cm, P < 0.05, Student's t-test). Our results revealed no statistical difference in head circumference, neck circumference, or neck length between each of the groupings. These results indicate a proportionately larger chest may be a protective factor in cervical spine fractures. PMID- 15124792 TI - A study of 13 cases of near-hanging presenting to an Accident and Emergency Department. AB - This paper reports 13 cases of near-hanging presenting to the Accident and Emergency (A + E) Department of a District General Hospital over a 52-month period. The purpose of the study was to analyse the epidemiology, methods, outcomes and complications of near-hanging. There were 13 cases in the study period, 12 of whom were male and 1 female. The mean age was 31 years. The most common method used was rope (five cases). The estimated mean duration of hanging was 5 min. Height of fall data was poorly recorded but only one fall was recorded as being more than 3 ft. Many had a very poor clinical picture on arrival of the ambulance crews: seven had a Glasgow Coma Score (GCS) of less than or equal to 8 on scene, of which five required intubation. Five patients required ventilation and a period of treatment on the Intensive Therapy Unit (ITU). There were no cervical spine (C-spine) injuries, nor neurological complications. There were two incidences of aspiration pneumonia. All of the patients made a full recovery. It is concluded that patients presenting with this injury go on to make a full recovery, despite an often dismal initial clinical picture. PMID- 15124793 TI - Marchetti nailing of humeral shaft delayed unions. AB - The purpose of this study is to present the results of Marchetti nailing in humeral shaft delayed unions. Fifteen humeral shaft delayed unions were treated with the Marchetti nail. The average age of patients was 52.3 years. All had been treated initially by conservative methods. Delay from trauma to surgery averaged 4.8 months. The nailing technique was retrograde, and reaming of the fracture focus was always performed. Bone graft was not added. Thirteen patients achieved union, in an average of 3.3 months. Two patients require further surgery to gain union. Shoulder and elbow motion was excellent in 10 patients, moderate in four and poor in one. The functional result was excellent in nine patients, good in two, fair in two, and poor in two. Marchetti nailing seems to be an acceptable alternative for humeral shaft delayed unions. PMID- 15124794 TI - Five years' clinical experience with the unreamed humeral nail in the treatment of humeral shaft fractures. AB - With the development of interlocking nail systems especially designed for the upper arm, standards for the operative treatment of humeral shaft fractures have appeared to change. The trumpet-like shape of the medullary cavity does not allow stable splinting with a nail alone, and therefore the bone--nail complex is commonly stabilized with interlocking bolts. Between June 1996 and June 2001, 51 fractures of the humeral shaft were treated operatively at the BG Unfallklinik Ludwigshafen with the unreamed humeral nail (UHN; Synthes). All nails were inserted by the retrograde technique. Ninety-five percent of the patients showed excellent or good shoulder function at follow-up examinations. For elbow function, 91.4% of the patients showed excellent or good results. Three out of four patients with poor elbow function had suffered from an additional injury to the brachial plexus; one patient developed heterotopic ossification. Intraoperative complications were: one iatrogenic lesion of the radial nerve, two intraoperative shaft fractures, one split at the insertion point, and one supracondylar fracture. As implants we used 7.5 mm nails in 36 cases and 6.7 mm nails in 15 cases. Among the 47 patients undergoing follow-up examinations, we found two cases of non-union. All patients were pain-free. Thirty-seven patients were very satisfied, six satisfied and four dissatisfied with the therapy. Decisive criteria for the use of a new implant are a high safety standard and simple reproducibility; these appear to be fulfilled by retrograde nailing of humeral fractures with the UHN. Interlocking nailing with the UHN enriches the range of therapeutic options for humeral shaft fractures. PMID- 15124795 TI - Tibial fractures associated with crush injuries to the soft tissues of the dorsal foot in children. AB - We retrospectively studied 15 children with tibial fractures associated with crush injuries to the soft tissues of the dorsal foot. The fractures, including six open fractures, were united with no complications within an average of 11.1 weeks. Wound closure to treat crush injuries of the dorsal foot was achieved using split- or full-thickness skin grafts in most patients. The outcomes of these grafts were acceptable, and all skin coverage was successful and remained viable with no breakdown. Extensor tendon injuries of the foot sustained by eight patients could not be sutured or repaired due to the nature of the injuries. However, the functional abilities of those injured tendons that could be sutured to surrounding tissues in a neutral position were acceptable, even though two patients had contracture of the toes that was problematic when wearing shoes. To manage crush skin injuries of the dorsal foot, split- or full-thickness skin grafts appear to provide a simple and convenient treatment strategy. In cases associated with extensor tendon injuries, suturing damaged extensor tendons to surrounding tissues represents another useful strategy with acceptable outcomes. PMID- 15124796 TI - Limited open reduction and Ilizarov external fixation in the treatment of distal tibial fractures. AB - The authors reviewed 31 distal tibial fractures (16 involving the tibial plafond) treated with Ilizarov external fixation. The study population was composed of 19 males and 12 females, with an average age of 54. The fractures were classified according to the AO classification: A1 (3), A2 (6), A3 (6), C1 (2), C2 (8), C3 (6). There were six open injuries. In 14 pilon cases, open reduction of the intra articular fragments and bone grafting via a limited incision was performed. Clinical follow-up averaged 28 months (range 18-42). All but one fracture united with an average healing time of 13.9 weeks. Nearly all patients with AO type A fracture had excellent or good functional scores. The 14 cases of AO C2 and C3 group had five (38%) good results, five (38%) fair results and three (24%) poor results. This method yielded results comparable with previous studies using open reduction and internal fixation. Twenty-nine percent of the patients had pin track infection, which remained the most important complication of this method. PMID- 15124797 TI - Fixation of tibial pilon fractures with percutaneous cannulated screws. AB - Closed reduction and internal fixation with percutaneous cannulated screws was performed on seven patients with closed pilon fractures. The mean follow-up was 30.6 months. They were assessed using a subjective scoring system. The average score was 90.8/100. This method of fixation avoids extensive soft tissue dissection and gives excellent results. To our knowledge, this method of closed reduction and stabilisation of pilon fractures solely by percutaneously inserted cannulated screws has not been previously reported. PMID- 15124798 TI - Differences in attitudes to analgesia in post-operative limb surgery put patients at risk of compartment syndrome. AB - Missed compartment syndrome may result in severe disability. Complete masking of pain, the cardinal symptom of compartment syndrome, may contribute to delayed or missed diagnosis. Scenarios reported in the literature as cases of delayed or missed compartment syndrome (due to analgesia) were included in a questionnaire. Each of the respondents was requested to indicate the preferred choice of post operative analgesia for each scenario. Significant differences were found between orthopaedic surgeons and anaesthetists regarding the preferred choice of post operative analgesia in clinical situations which have been previously shown to be associated with a high risk of compartment syndrome. Use of analgesic methods which impair the ability to detect patients with abnormal levels of pain or unusual demands for analgesia may place such patients at risk of the devastating sequelae of a missed compartment syndrome. The differences in attitudes to local and regional nerve blockade between orthopaedic surgeons and anaesthetists suggests that some anaesthetists may be exposing patients to the risk of missed compartment syndrome following extremity surgery. PMID- 15124799 TI - Does patient controlled analgesia delay the diagnosis of compartment syndrome following intramedullary nailing of the tibia? AB - We report on four cases in which the diagnosis of compartment syndrome was delayed by the administration of patient controlled analgesia (PCA) following intramedullary nailing of tibial shaft fractures. We believe that this poses a diagnostic problem and can lead to lasting sequelae as decompression is delayed. We recommend extra vigilance with the use of PCA in patients with intramedullary nailing following tibial shaft fractures. PMID- 15124800 TI - Hip disarticulation--the evolution of a surgical technique. AB - Introduced in the 18th century, hip disarticulation was considered to be one of the most radical operations performed for trauma or disease of the lower limb. The high morbidity and mortality associated with it ensured that it was a rarely performed procedure. It is fortunate that it remains extremely uncommon to the present day. Since the first successful hip disarticulation was described, a number of important advances have occurred. General medical care has improved dramatically and the development of anaesthesia, analgesics, antibiotics and blood transfusions has resulted in greatly decreased morbidity associated with this dramatic operation. This review on the history of hip disarticulation outlines the surgical evolution of the operation, the indications for its use and the techniques used. It draws on the early experiences and preferred techniques of the surgeons of the 19th century, with some discussion on the methods employed to reduce intraoperative haemorrhage. Further development of techniques in the 20th century is also described together with discussion on the evolution of hindquarter amputation. PMID- 15124801 TI - Prevention of posttraumatic hypoxaemia in isolated lower limb long bone fractures with a minimal prophylactic dose of corticosteroids. AB - The efficacy of a minimum dose of methylprednisolone for the prevention of posttraumatic hypoxaemia and fat embolism syndrome (FES) was prospectively studied in 87 patients with isolated, closed or grade I open, femoral and tibial fractures. On admission, the patients were randomly allocated either to a control group given placebo (40 patients) or to a methylprednisolone-treated group (47 patients). A total dose of 6 mg/kg BW methylprednisolone (SoluMedrol, Upjohn) was administered intravenously, divided in six equal doses at 8 h intervals. Six patients (12.8%) in the control group and one patient (2.5%) in the trial group developed FES (P = 0.079) but the difference is not statistically significant. Twenty-four hours after admission, the steroid-treated patients displayed statistically significant higher p(O2) values compared to the control group (P = 0.035) and this difference persisted on the second and the third post-admission day as well (P = 0.008). No corticosteroid-related side-effects were noticed in any of the patients during hospitalisation. Our results support the prophylactic administration of methylprednisolone in small dosage to prevent posttraumatic hypoxaemia and probably FES in patients with isolated lower limb long bone fractures, especially when early fracture stabilisation is not possible. PMID- 15124802 TI - Extensive disruption of abdominal wall musculature with intestinal prolapse, a rare injury caused by seat belt. PMID- 15124804 TI - Spontaneous rupture of a normal spleen. PMID- 15124805 TI - Pathological fracture of the distal humerus treated with Nancy nails. PMID- 15124803 TI - Injuries to the colon from blast effect of penetrating extra-peritoneal thoraco abdominal trauma. AB - Although rare, blast injury to the intestine can result from penetrating thoraco abdominal extra-peritoneal gunshot (and shotgun) wounds despite the absence of injury to the diaphragm or to the peritoneum. Injuries of the spleen, small intestine and the mesentery by this mechanism have been previously reported in the world literature. This paper reports the first two cases of non-penetrating ballistic trauma to the colon. PMID- 15124806 TI - Fixion--an inflatable or deflatable nail? PMID- 15124807 TI - Thyroid transection with spinal injury following blunt trauma in a child. PMID- 15124809 TI - Thick film traps with an irregular film. Preparation and evaluation. AB - A new method for preparation of sorbent-based ultra-thick film traps for concentration of trace volatile components from gaseous matrices is described. The procedure is based on blowing a prepolymer (polydimethylsiloxane) through a capillary tube, forming an irregular film of stationary phase. Subsequently, the prepolymer is immobilized in a few seconds by heating to 200 degrees C. Evaluation of the performance of the new traps showed that the loss of efficiency, compared to regular smooth film traps is only on the order of 20-30%. In terms of breakthrough volume, this loss in performance is rather insignificant. The technology is extremely simple and allows a rapid and cheap production of a large number of ultra-thick film traps, even in non-specialized laboratories. The method can be applied to any type of cross-linkable stationary phase, thereby expanding the scope of sorbent-based trapping and preconcentration concept. Many applications are anticipated in trace and ultra-trace analysis in a wide range of fields, such as environmental chemistry, polymers, food and process analysis. PMID- 15124808 TI - Blast injury caused by a booby-trapped cellular phone. AB - Terrorists are recently using cellular phones to remotely detonate bombs. A patient was injured while assembling a bomb connected to a cellular phone. The patient sustained combined injury to the head and to the dominant hand which held the phone. Amputation of the hand was required, the facial injuries were reconstructed. The characteristics of this unusual type of injury are described and compared to injuries caused by other bombs and explosive devices. PMID- 15124810 TI - Chiral separation and modeling of the three-chiral-center beta-blocker drug nadolol by simulated moving bed chromatography. AB - Nadolol, a beta-blocker used in the management of hypertension and angina pectoris, has three chiral centers and is currently marketed as an equal mixture of its four stereoisomers. Resolution of three of the four stereoisomers of nadolol was obtained previously by HPLC, with a complete separation of the most active enantiomer (RSR)-nadolol, on a column packed with perphenyl carbamoylated beta-cyclodextrin (beta-CD) immobilized onto silica gel. In this study, continuous separation of the target enantiomer of (RSR)-nadolol from its racemic mixture (which is a ternary mixture in the chromatographic system) was studied by non-linear SMB chromatography. Different regions of (2, 3) and (1, 2) complete separation regime were determined in the (m2, m3) region and the effect of non linearity such as overall feed concentration and component composition on the separation performances was investigated. A direct simulation approach has been proposed to simulate the SMB separation performance for the pseudo-binary mixture of nadolol. The simulation was conducted on the basis of a shortcut method constituted only of the weak-key and strong-key components. The performance of the cyclic steady-state behavior of the SMB unit was predicted reasonably well. It was also discussed quantitatively that the complete separation region obtained from the shortcut method is a subset of the true complete separation region and the optimal separation conditions obtained differed slightly from the "true" separation. PMID- 15124812 TI - Quantification from highly drifted and overlapped chromatographic peaks using second-order calibration methods. AB - For determining low levels of pesticides and phenolic compounds in river and wastewater samples by high performance liquid chromatography (HPLC), solid phase extraction (SPE) is commonly used before the chromatographic separation. This preconcentration step is not necessarily selective for the analytes of interest and it may retain other compounds of similar characteristics as well. In this case, we present, humic and fulvic acids caused a large baseline drift and overlapped the analytes to be quantified. The inaccurate determinations of the area of the peaks of these analytes made it difficult to quantify them with univariate calibration. Here we compare three second-order calibration algorithms (generalized rank annihilation method (GRAM), parallel factor analysis (PARAFAC) and multivariate curve resolution-alternating least squares (MCR-ALS)) which efficiently solve this problem. These methods use second-order data, i.e., a matrix of responses for each peak, which is easily obtained with a high performance liquid chromatography-diode array detector (HPLC-DAD). With these methods, the area does not need to be directly measured and predictions are more accurate. They also save time and resources because they can quantify analytes even if the peaks are not resolved. GRAM and PARAFAC require trilinear data. Biased and imprecise concentrations (relative standard deviation, %R.S.D. = 34) were obtained without correcting the time-shift. Hence, a time-shift correction algorithm to align the peaks was needed to obtain accurate predictions. MCR-ALS was the most robust to the time-shift. All three algorithms provided similar mean predictions, which were comparable to those obtained when sulfite was added to the samples. However, the predictions for the different replicates were more similar for the second-order algorithms (%R.S.D. = 3) than the ones obtained by univariate calibration after the sulfite addition (%R.S.D. = 13). PMID- 15124811 TI - Chromatographic impulse response technique with curve fitting to measure binary diffusion coefficients and retention factors using polymer-coated capillary columns. AB - The theoretical basis of a Gaussian-like approximate solution was applied to a chromatographic impulse response technique with curve fitting for measuring binary diffusion coefficients and retention factors using a polymer-coated capillary column. The formulae were derived for evaluating both the accuracy of the approximate solution and the sensitivity of the parameters. The validity of the solution also was confirmed experimentally for pulse injection of phenol in acetone into supercritical carbon dioxide flowing at 313.15 K and 11.6-28.6 MPa. Potential sources for experimental errors of the method are discussed. PMID- 15124813 TI - Transforming chiral liquid chromatography methodologies into more sensitive liquid chromatography-electrospray ionization mass spectrometry without losing enantioselectivity. AB - LC-electrospray ionization (ESI) MS conditions were optimized for the individual chiral separation of 19 compounds of pharmaceutical interest using the macrocyclic glycopeptide-based chiral stationary phases in both polar organic and reversed-phase modes (RPM). The influence of mobile phase composition and MS additive type on sensitivity was investigated for all classes of compounds tested. Compounds with amine or amide groups were efficiently separated, ionized, and detected with the addition of 0.1% (w/w) ammonium trifluoroacetate to the solvent system in either the reversed-phase or polar organic mode (POM). Macrocyclic glycopeptide coupled column technology was initially used to screen all chiral compounds analyzed. Baseline resolution of enantiomers was then achieved with relatively short retention times and high efficiencies on Chirobiotic T, Chirobiotic V or Chirobiotic R narrow bore chiral stationary phases. The polar organic mode offered better limits of detection (as low as 100 pg/ml) and sensitivity over reversed-phase methods. An optimum flow-rate range of 200-400 microl/min was necessary for sensitive chiral LC-ESI-MS analysis. PMID- 15124814 TI - Chirality determination of unusual amino acids using precolumn derivatization and liquid chromatography-electrospray ionization mass spectrometry. AB - Unusual amino acids such as beta-methoxytyrosine (beta-MeOTyr), allo-threonine (allo-Thr) and allo-isoleucine (allo-Ile) were derivatized with N-alpha-(2,4 dinitro-5-fluorophenyl)-L-alaninamide (FDAA), 2,3,4,6-tetra-O-acetyl-beta-D glucopyranosyl isothiocyanate (GITC), (S)-N-(4-nitrophenoxycarbonyl)phenylalanine methoxyethyl ester (S-NIFE), or o-phthalaldehyde/isobutyryl-L-cysteine (OPA IBLC), and then separated via reversed-phase high-performance chromatography followed by UV and electrospray ionization mass spectrometry detection. FDAA generally showed the highest enantioselectivity but the lowest sensitivity among the chiral derivatizing agents (CDAs) investigated. The detection limit of FDAA derivatized amino acids was in the low picomolar range. Although the enantioselectivity of FDAA derivatives was generally quite high, its selectivity among beta-MeOTyr isomers was poor. The best separation of beta-MeOTyr stereoisomers was achieved with S-NIFE. Due to the complex relationships between the investigated CDAs, stereochemical analyses using a combination of two or more of the CDAs gave the most reliable results for a given separation problem. In general, the methods described are selective and reliable, and are being applied to the analysis of unusual amino acids as they occur in marine peptides. PMID- 15124815 TI - Post-column addition as a method of controlling triacylglycerol response coefficient of an evaporative light scattering detector in liquid chromatography evaporative light-scattering detection. AB - The non-linear response is generally the main limitation to the general quantitative use of evaporative light-scattering detection (ELSD). In the particular case of triacylglycerol (TG) analysis, we present a preliminary paper dealing with the use of post-column additives as a means of monitoring the response of such a detector. As TG can form molecular association complexes (ligand-ligate associations) with either cholesterol, urea or silver nitrate, we report the influence of the concentration of each of these chemical compounds in the liquid phase directed towards the ELSD system. The results show that the response coefficient b of the calibration curve either decreases from 1.25-1.30 to 0.51 or increases from 1.25-1.30 to 1.78 according to the nature and concentration of post-column additive. The use of cholesterol as additive, at a discrete concentration, may lead to a linear response curve (b = 1), i.e. to the direct proportionality of ELSD response versus the TG concentration, making quantitative analysis of such solutes easier. On the other hand, to improve sensitivity, the addition of silver nitrate may be chosen for an increase in b value. PMID- 15124816 TI - Oxidative effects induced by dediazoniation of the p-hydroxybenzenediazonium ion in a neutral aqueous medium. AB - The toxicity of arenediazonium ions is believed to result from the appearance of very reactive compounds during the dediazoniation process. In the case of the p hydroxybenzenediazonium ion (PDQ), radical species generated during dediazoniation could potentially initiate lipid peroxidation. The data obtained in spectrophotometric experiments suggest that an interaction between PDQ and linoleic acid (LA) gives rise to the characteristic absorption of oxidized products deriving from LA, both in the presence and absence of a mixed micellar medium containing the surfactant Tween 20 (Tw20). Spectroscopic evidence also clearly points to the interference of these processes in the dediazoniation of PDQ. Analysis by reverse-phase, high-pressure liquid chromatography (HPLC) confirms that the decomposition of PDQ in a mixed micellar medium induces the peroxidation of both LA and methyl linoleate (MEL), thus causing the appearance of peaks characteristic of dienic conjugated hydroperoxides. The same products are observed after interaction between LA and the water-soluble 2,2'-azobis (2 amidinopropane), a frequently used initiator of lipid peroxidation. The proportion of isomers produced during the peroxidation process agrees well with that reported for reactions mediated by free radicals. A further chromatographic analysis of the decomposition of PDQ in the presence of 2-methylcyclohexa-2,5 diene-1-carboxylic acid (CHD) shows that phenol and quinone are the main products of the reaction. These results are discussed on the understanding that aryl and peroxyl radicals abstract a hydrogen atom from CHD, in accordance with our general scheme for PDQ dediazoniation described in a previous publication. PMID- 15124817 TI - Evaluation of fast gas chromatography and gas chromatography-mass spectrometry in the analysis of lipids. AB - Fast and conventional gas chromatography (GC) techniques were applied to nine different lipidic matrices (butter, lard, tallow, and peanut, corn, sunflower, soya, olive, menhaden oils). Simultaneous methylic transesterification was performed on all samples prior to GC analysis. Several practical aspects concerning high speed analysis were investigated, such as the great increase in linear velocity, the use of fast temperature ramps, column sample capacity and detection systems. Analytical results showed certain losses in resolution, balanced by a consistent reduction in analysis time. The actual time savings were variable (60-70 min) as they were dependent on the complexity of the sample while the speed enhancement factor was equal to 10.5. Peak identification was achieved by means of different information sources, such as fast GC-mass spectrometry (MS), linear retention indices and comprehensive two-dimensional (2D) gas chromatography group patterns. The method developed was shown to be applicable in routine applications on complex natural samples. PMID- 15124819 TI - Evaluation of a pulse-discharge helium ionisation detector for the determination of neon concentrations by gas chromatography. AB - A pulse-discharge helium ionisation detector, PDHID (Valco, PD-D2-I) with sample introduced to the discharge zone is shown to be applicable for reliable determinations of neon by gas chromatography. The detection level of 80 pg was obtained, but the dependence between detector response and neon mass was non linear. However, for the discharge gas doped with 33 ppm of neon, a linear response to the neon mass up to 10(-5) g and the detection level of 0.5 ng were obtained. The method can be used for measuring neon concentrations in groundwater systems for hydrogeological purposes. PMID- 15124818 TI - Development and validation of a highly sensitive gas chromatographic-mass spectrometric screening method for the simultaneous determination of nanogram levels of fentanyl, sufentanil and alfentanil in air and surface contamination wipes. AB - A highly sensitive gas chromatographic-mass spectrometric (GC-MS) analytical method for the determination of the opioid narcotics fentanyl, alfentanil, and sufentanil in industrial hygiene personal air samples and surface contamination wipes was developed and comprehensively validated. Sample preparation involved a single step extraction of the samples with methanol, fortified with a fixed amount of the penta-deuterated analogues of the opioid narcotics as internal standard. The GC-MS analytical procedure using selected ion monitoring (SIM) was shown to be highly selective. Linearity was shown for levels of extracted wipe and air samples corresponding to at least 0.1-2 times their surface contamination limit (SCL) and accordingly to 0.1-2 times their time weighted average occupational exposure limit (OEL-TWA) based on a full shift 9601 air sample. Extraction recoveries were determined for spiked air samples and surface wipes and were found to be quantitative for both sampling media in the entire range studied. The air sampling method's limit of detection (LOD) was determined to be 0.4 ng per sample for fentanyl and sufentanil and 1.6 ng per sample for alfentanil, corresponding to less than 1% of their individual OEL for a full shift air sample (9601). The limit of quantification (LOQ) was found to be 1.4, 1.2, and 5.0 ng per filter for fentanyl, sufentanil, and alfentanil, respectively. The wipe sampling method had LODs of 4 ng per wipe for fentanyl and sufentanil and 16 ng per wipe for alfentanil and LOQs of respectively, 14, 12, and 50 ng per wipe. The analytical intra-assay precision of the air sampling and wipe sampling method, defined as the coefficient of variation on the analytical result of six replicate spiked media was below 10 and 5%, respectively, for all opioids at all spike levels. Accuracy expressed as relative error was determined to be below 10%, except for alfentanil at the lowest spike level (-13.1%). The stability of the opioids during simulated air sampling was investigated. For fentanyl and sufentanil a quantitative recovery was observed at all spike levels, while for alfentanil recoveries ranged from 60.3 to 85.4%. When spiked air samples were stored at ambient temperature and at -15 degrees C quantitative recovery was found for fentanyl and sufentanil after 7 and 14 days. For alfentanil a slight loss seemed to occur upon storage during 7 days, being more explicit after 14 days. Ambient storage of spiked wipes seemed to lead to significant losses of all opioids studied, yielding recoveries of 37.7-88.3%. Upon storage of similar wipes at -15 degrees C a significantly higher recovery was found ranging from 77.3 to 88.3%. The developed analytical and sampling procedures have been recently applied in an explorative field study of which the results of surface contamination wipe sampling are presented in this paper. To our knowledge, this is the first study addressing the development and validation of analytical procedures for the assessment of external occupational exposure to potent opioid narcotics. PMID- 15124820 TI - Separation of acidic and basic compounds in capillary electrochromatography with polymethacrylate-based monolithic columns. AB - Methacrylate-based monolithic columns with electroosmotic flow (EOF) or very weak EOF are prepared by in situ copolymerization in the presence of a porogen in fused-silica capillaries pretreated with a bifunctional reagent. Satisfactory separations of acidic and basic compounds on the column with EOF at either low or high pH are achieved, respectively. With sulfonic groups as dissociation functionalities, sufficient EOF mobility still remains as high as 1.74 x 10(-4) cm2 s(-1) V(-1) at low pH. Under this condition, seven acidic compounds are readily separated within 5.7 min. Moreover, at high pH, the peak shape of basic compounds is satisfactory without addition of any masking amines into running mobile phase since the secondary interaction between the basic compounds and the monolithic stationary phase are minimized at high pH. Reversed-phase mechanism for both acidic and basic compounds is observed under investigated separation conditions. In addition, possibilities of acidic and basic compound separations on a monolithic column with extremely low EOF are discussed. PMID- 15124821 TI - Isotachophoretic separation of alkylsulfonates and determination of methanesulfonic acid as main component and as trace component in pharmaceutical drug substances. AB - Alkylsulfonates from methanesulfonic acid to decanesulfonic acid were separated by isotachophoresis with conductivity detection in a common electrolyte system at pH 4.8. The electrolyte system consisted of 10 mM HCl buffered with epsilon aminocaproic acid (pH 4.8) and 0.1% methylhydroxyethylcellulose (MHEC) acting as the leading electrolyte. The terminating electrolyte was 20 mM caproic acid also containing 0.05% MHEC. Current settings of 250 microA for the first and 50 microA for the second capillary were applied. On one hand, the method was applied to the determination of the content of methanesulfonate as the salt forming agent (mesilate) in a recently registered drug substance. The results obtained by ITP were compared with an orthogonal titration method. On the other hand, due to the column-coupling configuration of the electrophoretic instrument, the method could be extended to the trace determination in the ppm range in order to monitor methanesulfonic acid as an impurity in a drug substance. The validation confirmed the linearity of the method between 1 and 10 mg/l, limits of detection and quantification below 1 mg/l, recovery rates from 92.4 to 95.4%, and repeatability with a R.S.D. of 3.8% (six runs with a 4 mg/l spiked sample). Finally, three batches of a newly produced drug substance could be checked for methanesulfonic acid giving results of below 0.0014% (concentration related to the drug substance). PMID- 15124822 TI - Development of a novel solid-phase extraction element for thermal desorption gas chromatography analysis. AB - A novel solid-phase extraction element is developed for sorptive enrichment of dilute analytes from liquid samples with high extraction efficiencies due to its larger amounts of polydimethylsiloxane (PDMS) absorbent than the conventional syringe type of solid-phase microextraction (SPME). The extraction element is made of titanium (Ti) open tubular tube (30 mm x 1.2 mm i.d. x 1.6 mm o.d.) coated with a chemically bonded layer of PDMS (500 microm in thickness). The extraction element combined with thermal desorption-gas chromatography-mass spectrometry using a pyrolysis-gas chromatography-mass spectrometry system was used to extract and analyze a typical herbicide, bethrodine in water samples over a concentration range from 2.5 to 2.5 x 10(4) ng/l. Thus obtained calibration curve showed good linearity for the tested whole concentration range with regression coefficient of 0.992. Detection limit of 0.5 ng/l level was achieved and the reproducibility of the measurements for bethrodine at 10 ng/l level was found to be fairly good with relative standard deviation below 7.5%. PMID- 15124823 TI - New hydrophilic polymeric resin based on 4-vinylpyridine-divinylbenzene for solid phase extraction of polar compounds from water. AB - A 4-vinylpyridine-divinylbenzene (VP-DVB) resin was synthesized to be used for on line solid-phase extraction process and it was tested for a group of polar compounds. The high specific surface area and the nitrogen content of the VP-DVB sorbent increased the interactions with the polar analytes in the preconcentration process. The sorbent enabled 100 ml of water to be concentrated with recoveries higher than 70% for several polar compounds (including phenol) except for oxamyl (55%) and methomyl (43%). The method was used to analyse water samples by liquid chromatography and UV detection. Linearity was good and detection limits were 0.1-0.2 microg l(-1) for all compounds. Several tap and river water and waste water treatment plant samples were analyzed; phenol and (4 chloro-2-methyl-phenoxy)acetic acid (MCPA) were tentatively determined in some samples. PMID- 15124824 TI - Determination of carnosine in feed and meat by high-performance anion-exchange chromatography with integrated pulsed amperometric detection. AB - Carnosine (beta-alanyl-L-histidine) is a dipeptide regarded as an important molecular marker of the presence of processed animal proteins including meat and bone meal in animal feed. For its identification and quantification a sensitive and selective method by high-performance anion-exchange chromatography coupled with integrated pulsed amperometric detection (HPAEC-IPAD) was developed. The assay is based on isocratic elution with 100 mM NaOH as the mobile phase. Interferences of real matrices were efficiently removed; carnosine could be determined at the concentration ranges 0.1-100 microM with a rather low detection limit of 0.23 ng. Unlike feeds for dogs and cats, no carnosine peak was observed in all examined feeds for ruminants. The good analytical characteristics allowed camosine determination down to 5 microg/g of feed. PMID- 15124825 TI - Determination of polybrominated diphenyl ethers in marine biological tissues using microwave-assisted extraction. AB - Growing concern on the environmental impact of polybrominated diphenyl ethers (PBDEs) has created the need for rapid and quality assured analytical methods to quantify PBDEs in a spectrum of matrix types. This study presents the first validated method for the quantification of major PBDE congeners (47, 99 and 100) in marine biological tissues using microwave-assisted extraction (MAE). The recovery of polychlorinated biphenyls and various organochlorine pesticides has also been ascertained. Analytical accuracy, precision, limits of detection and cleanup efficiency were evaluated for PBDE congeners, and empirical data justifies the use of MAE for the extraction and analysis of PBDEs in biological matrices. MAE was also compared to Soxhlet extraction efficiency for PBDEs in the standard reference materials SRM2978 and SRM1588a and gave comparable results (<15% variation). PMID- 15124826 TI - Progress in life sciences and translation into clinical application. PMID- 15124827 TI - Translating knowledge into practice in the "post-genome" era. AB - The Human Genome Project is "completed", but it is only a beginning in the understanding of genomic structure and function. A "human phenome project" is waiting in the wings. The complexity involving a phenotype can be glimpsed, for example, if one enquires into the relationships between mutant phenylalanine hydroxylase (PAH) genotypes and the clinical disorders called PKU/Hyperphenylalaninemia-so called lessons from PKU genotypes and phenotypes. Since genomes speak biochemistry, not phenotype (said RHA Plasterk), for genomics to penetrate medicine, biochemistry and biology must be allies. The ideal translators and ambassadors of the knowledge that must cross the gap between laboratory and bedside are the clinician scientists; restoration of that attenuated community of colleagues is a necessary step in the implementation of genomic medicine. PMID- 15124828 TI - A classification of end-of-life decisions in neonates and infants. AB - AIM: Valid and reliable data are needed to gain insight into the decisions of paediatricians concerning the end of life of newborns and infants. Such data could throw light on the poorly documented aspects of medical practice regarding these end-of-life decisions (ELDs). In this article a classification of ELDs is developed. METHODS: The classification is developed from knowledge generated from large-scale epidemiological studies on ELDs. RESULTS: This classification sets out from considerations of the various possible medical end-of-life procedures for newborns and infants and from important ethical aspects of the decision making process. From both ethical and legal viewpoints the life-shortening intention of the physician is a significant factor for the qualification of ELDs. Furthermore, the consultation of the parents is an important factor in absence of the possibility of the patient's self-determination. CONCLUSION: This classification has been devised with the aim of making reliable and valid descriptions of both the incidence and nature of ELDs in this specific population of newborns and infants. PMID- 15124829 TI - Are our children fit or fat? AB - Overweight and obesity are increasing in Swedish boys and girls aged 7-16 y. The fitness of 16-y-old Swedish boys in 2001 was approximately 10% worse than in 1987. CONCLUSION: It is important to start the prevention of atherosclerosis and type 2 diabetes early. The number of children with physical inactivity and obesity should be as low as possible. PMID- 15124830 TI - Prevention of neonatal cold injury in preterm infants. AB - In many parts of the world neonatal cold injury still contributes to neonatal death in preterm or growth-retarded neonates. Kangaroo Mother Care might improve the situation and is the only effective, affordable and available method to prevent neonatal hypothermia in most developing countries. In developed countries, highly sophisticated incubators and radiant warmers are available but there is increasing concern about the long-term effects of high levels of sound, disturbing light and unplanned procedures which can have a bearing on the use of these tools. A controlled clinical trial of cot-nursing with a heated, water filled mattress is presented in this issue of Acta Paediatrica. CONCLUSION: The results from the new trial and the recently published systematic reviews by the Cochrane Library of technologies to provide warmth to preterm infants in modem neonatal intensive care units, makes it possible to suggest the place of available tools: incubators, radiant warmers, heated water-filled mattresses and care in skin-to-skin contact. PMID- 15124831 TI - Detection of Mycobacterium tuberculosis in gastric aspirate and sputum collected from Ethiopian HIV-positive and HIV-negative children in a mixed in- and outpatient setting. AB - AIM: To investigate, through a prospective study, the detection rate of Mycobacterium tuberculosis in sputa and gastric aspirate from tuberculous children in a low-income country with high prevalence of tuberculosis and an increasing HIV epidemic. METHODS: Gastric aspirates and/or sputum samples were collected from 355 children with pulmonary tuberculosis as follows: from 136 children under 5 y only gastric aspirate was taken, for 159 children aged 5 to 9 y both methods were used, and for 60 children over 10 y only sputum was analysed. The diagnosis of tuberculosis was based on clinical data, tuberculin test and chest radiography. All children were tested for HIV infection. RESULTS: Direct microscopy for acid-fast bacilli was positive for 55 (15%) and mycobacterial culture for 183 (52%) children. The proportion of positive cultures was similar in all age groups. Among the 5 to 9 year-old children who could produce a sputum sample, sputum gave just as good culture yield of M. tuberculosis as gastric aspirate. Of the clinical or radiological findings only weight loss was associated with a higher yield. Repeat gastric aspirate increased the culture yield by 6%. Mycobacterial culture from HIV-positive children gave lower yield compared with HIV-negative children. CONCLUSION: Our data suggest that one gastric aspirate for children less than 6 y and three sputum samples for the older children collected at an outpatient TB clinic, is enough to provide a close to 50% yield of M. tuberculosis available for culture and further analyses. However, with an increasing prevalence of HIV, this detection rate may be reduced. PMID- 15124832 TI - Respiratory outcome in school-aged, very-low-birth-weight children in the surfactant era. AB - AIM: To assess respiratory outcome and its predictors during the surfactant era in very-low-birth-weight (VLBW, birth weight <1500g) schoolchildren with and without bronchopulmonary dysplasia (BPD). METHODS: At 7-8 years of age, 34 VLBW children with BPD diagnosed at a postnatal age of 28 d underwent flow-volume spirometry, metacholine challenge, bronchodilatation test, whole body plethysmography and diffusion capacity measurement. Fourteen of them had not recovered from BPD by a corrected gestational age of 36 wk (sBPD subgroup). The age- and sex-matched control groups comprised 34 VLBW cases without BPD and 34 term children. RESULTS: Current respiratory symptoms in contact with cold air and/or upon exercise were reported in one-third of the VLBW children. Only half of the symptomatic VLBW cases without BPD had inhaled medications. Compared with term controls, the BPD cases had lower forced expiratory volume in 1 s (FEV1), higher ratio of residual volume to total lung capacity and higher airway resistance. Lower FEV1 and specific conductance were found in the sBPD subgroup compared to both control groups. Additionally, their vital capacity was lower than in term controls. A higher rate of bronchial hyper-reactivity and lower diffusion capacity of the lungs were detected in VLBW as against term cases. Low birth weight, long duration of oxygen therapy, low socio-economic status and exposure to animal dander emerged as predictors of poorer respiratory outcome. CONCLUSION: In the surfactant era, birth weight, neonatal respiratory morbidity, as well as later environmental factors appear to affect the respiratory outcome of VLBW children. However, careful pulmonary follow-up of all VLBW children seems to be indicated regardless of the severity of neonatal respiratory problems. PMID- 15124833 TI - Cytokines in the gastric mucosa of children with Helicobacter pylori infection. AB - AIM: Few studies have looked at the cytokine profile in gastric mucosa in children with Helicobacter pylori infection. This study investigated cytokines and their effects on histological abnormalities in the gastric mucosa of children with H. pylori infection. METHODS: The levels of interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and IL-8 proteins were measured in biopsy specimens from the gastric antrum and corpus of children with H. pylori infection, and related to inflammatory cell infiltrations. RESULTS: The antral and corporal mucosal levels of IFN-gamma and IL-8 proteins were significantly higher in children with H. pylori infection than in uninfected children, but there was no such difference in the levels of IL-4 protein. The antral mucosal level of IL-8 protein was significantly higher than the corporal mucosal level of IL-8 protein in the infected children. Inflammatory cell infiltration was significantly higher in the infected children than in the uninfected children, but there were no significant correlations between mucosal cytokine levels and inflammatory cell infiltrations. CONCLUSION: The results suggest that the predominant Th1 cytokine response and enhanced IL-8 production in the mucosa may be involved in the gastric inflammation seen in children infected with H. pylori, as well as in adult patients. PMID- 15124834 TI - Elemental versus polymeric enteral nutrition in paediatric Crohn's disease: a multicentre randomized controlled trial. AB - AIM: To compare the efficacy and safety of an elemental and a polymeric diet as the primary therapy for active Crohn's disease in children. METHODS: In a randomized, non-blind, multicentre, controlled trial in Sweden, 16 children with Crohn's disease received Elemental 028 Extra (E028E) and 17 Nutrison Standard (NuS). Remission rates (Paediatric Crohn's Disease Activity Index (PCDAI) < 10 or a PCDAI decrease of 40% or 15 points of initial level) were compared at 6 wk. RESULTS: There was no significant difference between the two groups in remission rate at 6 wk (intent-to-treat analysis): E028E 11/16 (69%) and NuS 14/17 (82%) (p = 0.438). There was no difference in the decrease in PCDAI and CDAI between patients treated with E028E and those treated with NuS from 0 to 6 wk. Patients treated with NuS gained significantly more weight than patients treated with E028E (+2.5 kg; 95% CI 0.9, 4.1; p = 0.004), this difference remained when adjusting for maximum caloric intake per kilogram bodyweight (+2.9 kg; 95% CI 1.4, 4.5; p = 0.001). Concomitant disease, complications and side effects were seen in 5/33 patients (pyelonephritis, pneumonia, intraabdominal abscess, perianal abscess and borborygmi). CONCLUSION: E028E and NuS did not differ in terms of remission rate. Patients treated with NuS gained more weight than patients with E028E. Polymeric diet may be superior to elemental diet in the treatment of paediatric Crohn's disease where the primary aim is to increase the patient's weight. PMID- 15124835 TI - Antenatal renal pelvis dilatation emphasizing vesicoureteric reflux: two-year follow-up of minor postnatal dilatation. AB - AIM: To describe the occurrence of foetal renal pelvis dilatation in an unselected population of pregnancies and to describe the clinical course in a subgroup of infants with minor renal pelvis dilatation (RPD). METHODS: During 1996-1999 the foetuses of 17850 consecutive pregnant women were scanned with ultrasonography (USG) in the 16th-19th gestational week and later in the pregnancy when indicated. RPD was defined as anterior-posterior pelvic diameter > or = 5 mm. After birth all infants were examined with USG twice. If any postnatal USG showed RPD > or = 10 mm, extended radiological investigation was performed. Infants with pelvis dilatation < 10 mm and no calyceal or ureteric dilatation were not investigated further and antibiotic prophylaxis was discontinued. The children were followed up at 2 y of age. RESULTS: 109 foetuses (0.6%) with RPD were identified. Postnatally, 94/109 were available for follow-up: 43 had postnatal dilatation > or = 10 mm and were investigated and treated according to clinical routine, while 51 had dilatation < 10 mm and were followed up after they had reached at least 2 y of age. No case of pyelonephritis was recorded. USG in 48/51 children was normal. CONCLUSION: The data indicate that infants with minor RPD do not constitute a risk group for renal morbidity and thus do not need antibiotic prophylaxis and extensive radiological examination. PMID- 15124836 TI - Clinical signs of heart failure are associated with increased levels of natriuretic peptide types B and A in children with congenital heart defects or cardiomyopathy. AB - AIM: To study whether natriuretic peptide types B (BNP) and A (ANP) reflect clinical signs of heart failure (CSHF) in children with congenital heart defects or cardiomyopathy resulting in different types of haemodynamic situations, such as pressure overload in coarctation of the aorta (CoA), volume overload in ventricular septal defect (VSD) or systolic dysfunction in dilated cardiomyopathy (DCM). METHODS: Blood samples for plasma P-BNP and P-ANP were taken before procedures during regular investigation from 26 children (9 CoA, 11 VSD and 6 DCM). The ordinary paediatric cardiologist performed the cardiac evaluation and the data were retrieved from medical charts. CSHF was considered positive if two of the following criteria were fulfilled: reduced physical capacity, feeding disorders, dyspnoea, tachypnoea, hepatomegaly and oedema. The statistical methods were non-parametric. RESULTS: 0/9 children with CoA, 5/11 with VSD and 6/6 with DCM had CSHF. In children with CSHF, P-BNP and P-ANP were higher, 263 ng l(-1) (range 47.5-1300) and 303 ng l(-1) (range 168-466), than in those without CSHF, 12.3 ng l(-1) (range 4.8-30.8) and 42.9 ng l(-1) (range 13.7-189), respectively (p < 0.001, Mann-Whitney U-test), irrespective of the diagnosis. The same relationship was also found in the group of children with VSD. CONCLUSION: Plasma levels of ANP and BNP increase in children with CSHF. This increase is seen irrespective of whether it is due to systolic dysfunction, as in children with DCM, or to a volume overload with a normal systolic function, as in children with VSD. PMID- 15124837 TI - Maternal predictors of subcutaneous fat in the term newborn. AB - AIM: To determine the relative influences of some maternal factors on skinfold thickness. The effects of age, parity, height, body mass index (BMI) and pregnancy weight gain (PWG) of the mother on the subscapular skinfold thickness (SST) of the newborn were estimated, and compared with their effects on birthweight (BW), crown-heel length (CHL) and head circumference (HC). METHODS: A sample of 13, 972 healthy, term singletons was selected at the Clamart Maternity Hospital (France). Stepwise regressions were used to determine the most predictive maternal factors for each parameter in the newborn. The respective effects of the mother's age and parity on each newborn dimension were tested by analysis of variance. RESULTS: The SST is a singular parameter, influenced by the mother's BMI and PWG, but not by her height. In contrast, the main predictor of BW, CHL and HC is the height of the mother, and to a lesser extent her PWG and BMI. Parity and maternal age have a smaller effect, except on SST, and essentially between the first and second pregnancies. CONCLUSION: These results clearly separate SST from other newborn dimensions. The skinfold thickness depends only on the nutritional status of the mother, while other dimensions are markedly influenced by the genetic background. This study is the first to demonstrate the singularity of skinfold thickness in newborn infants as a marker of the mother's nutritional status. PMID- 15124838 TI - Cot-nursing using a heated, water-filled mattress and incubator care: a randomized clinical trial. AB - AIMS: To evaluate the thermal responses and weight gain in preterm infants nursed in a cot on a heated, water-filled mattress (HWM) compared with infants receiving care in an air-heated incubator and to compare mothers' stress, anxiety levels and perceptions of their infants in the two groups. METHODS: Stable preterm infants weighing 1300 to 1500 g were enrolled, being randomly allocated to either the study group (n = 41) receiving care in a cot on an HWM, or the control group (n = 33) receiving incubator care. The mean daily body temperature and episodes of cold stress and hyperthermia were recorded. Weight gain (g kg(-1) body weight d(-1)) was also calculated. The mothers completed questionnaires on their perceptions of their infants, and their anxiety and stress levels before randomization, and 2-3 wk later during the trial. RESULTS: The mean body temperature was similar for the first week of the trial (study group 36.9 degrees C vs controls 36.9 degrees C). There were no significant differences in the incidence of cold stress, while more hyperthermic episodes were seen in the study group (p = 0.03). There were no significant differences in weight gain during the first (study group 21.4 g vs controls 19.6 g) or second weeks of the trial (study group 20.5 g vs controls 19.2 g). Neonatal morbidity did not differ between the groups. There were no differences in mothers' perceptions of their babies, or feelings of stress or anxiety. CONCLUSION: There were no differences between infants cot-nursed on an HWM and those receiving incubator care, with the exception of episodes of high temperature. The results suggest that the HWM may be used safely for low-weight preterm infants. PMID- 15124839 TI - Evaluation of an amikacin loading dose for nosocomial infections in very low birthweight infants. AB - AIM: To develop a simplified amikacin dosage regimen for nosocomial infections in preterm infants including a loading dose in order to achieve therapeutic Maximum Serum Concentrations early in the course of therapy. METHODS: Open, non comparative study during November 2000 to April 2001. The modified amikacin dosing and monitoring protocol included a loading dose of 10 mg/kg in the first week of life, followed by a maintenance regimen of 7.5 mg/kg every 24 h. After the first week of life the corresponding doses were 17 mg/kg (loading) and 15 mg/kg (maintenance). A peak level was measured 30 min after the second dose, a trough level immediately before the third dose. RESULTS: Twenty-five very low birthweight infants (median birthweight 739 g, median gestational age 25 wk) who had 34 episodes of amikacin treatment were included in the analysis. Median amikacin peak and trough values were 37.1 micromol/l and 6.3 micromol/l, respectively. Twenty-nine of all peak levels (85%) and 30 of all trough levels (88%) were within the targeted range of >35 micromol/l and <8.5 micromol/l, respectively. All patients with elevated trough levels were of extremely low birthweight and were born in the 24th week of gestation. Hearing evaluations were performed in 17 of 19 surviving infants at discharge home, all of which gave normal results. CONCLUSION: The new amikacin dosing protocol yielded targeted peak and trough concentrations in a high percentage of very low birthweight infants with nosocomial infection after the first week of life. Our simplified dosage regimen achieved acceptable serum concentrations in all birthweight and gestational age groups, with the exception of extremely low birthweight infants weighing less than 700 g and/or with a gestational age of 24 wk or less. Only limited information can be gained from our data regarding the use of amikacin during the first week of life. PMID- 15124840 TI - Long-term follow-up of otherwise healthy term infants with marked hyperbilirubinaemia: should the limits of exchange transfusion be changed in Turkey? AB - AIM: To evaluate prospectively non-haemolytic term infants with marked hyperbilirubinaemia treated by phototherapy only for evidence of bilirubin toxicity at 2-6 y of age, and to determine the suitability for Turkish children of the exchange transfusion limits recently reported by the American Academy of Pediatrics. METHODS: The study group included a total of 30 children, aged 2-6 y, who had developed marked hyperbilirubinaemia (20-24 mg dl(-1), 342-410 micromol l(-1)) during the newborn period (gestational age >37 wk, birthweight >2500 g) and were treated without exchange transfusion because intensive phototherapy, instituted during the preparations for exchange transfusion, was successful in decreasing their serum bilirubin levels. The control group consisted of 30 children of the same age group without clinical jaundice in the newborn period. Physical and neurological examinations, brainstem auditory-evoked potentials (BAEPs) and developmental tests for Turkish children were performed in both the study and control children. RESULTS: There was no difference between the groups with regard to mean BAEP latencies and developmental scores. None of the infants had hearing loss, developmental delay or abnormal neurological findings. CONCLUSION: The results suggest that successful intensive phototherapy without exchange transfusion in otherwise healthy term newborn infants with marked hyperbilirubinaemia (20-24 mg dl(-1), 342-410 micromol l(-1)) might not increase the risk of bilirubin brain injury and that the conventional limit of 20 mg dl( 1) (342 micromol l(-1)) could be changed to 22-24 mg dl(-1) (376-410 micromol l( 1)) for healthy term infants in Turkey. These limits, however, address only infants who do not have haemolytic disease, and the data are not sufficient to draw conclusions on the safety of even higher bilirubin levels (i.e. >24 mg dl( 1), 410 micromol l(-1)) in this population. PMID- 15124841 TI - Association between low Apgar score and neonatal cholestasis. AB - AIM: To investigate the association between low Apgar score and the development of cholestasis. METHODS: Seventy-seven cholestatic infants, all referred to our tertiary centre and born between 1987 and 1996 were studied. Twenty-eight patients had biliary atresia (BA), 36 had various intrahepatic disorders and for 13 patients the aetiology of the cholestasis was unknown. Data on gestational age, mode of delivery, Apgar score and birthweight for the cholestatic infants and 1,118,270 control subjects born during the same time period were obtained from the Swedish Medical Birth Registry. If the Apgar score of the cholestatic patient was <7 at 1 min and/or <9 at 5 min and/or <9 at 10 min of age the available medical records were reviewed for signs of neonatal distress. RESULTS: Five cholestatic patients, all of them premature, fulfilled the Apgar criteria. For two of them the low Apgar score and need for immediate resuscitation were explained by major surgical problems. The other three patients, two with biliary atresia (BA) and one with Alagille syndrome, had clinical signs of neonatal distress. The incidence of low Apgar score in BA patients was 7% and in cholestatic patients without known aetiology 0%, neither figure differing significantly from that of the of the control group (2.6%). CONCLUSION: Low Apgar score is not more common in any of the cholestatic groups than in the general Swedish population of newborns. We suggest that aetiological associations other than low Apgar score need to be considered in infants with cholestasis of unknown cause. PMID- 15124842 TI - Human immunodeficiency virus infection, diarrheal disease and sociodemographic predictors of child growth. AB - AIM: To compare growth patterns between human immunodeficiency virus (HIV) infected and -uninfected preschool children. To examine the associations between diarrheal and respiratory infections, sociodemographic factors and growth. METHODS: A longitudinal study was conducted among 524 children who were 6-60 mo of age at recruitment. Information on sociodemographic characteristics was collected at baseline from the caregiver. Hemoglobin, malaria infection and HIV status of the children were assessed from a blood sample. Monthly height (length if <24 mo) and weight measurements were obtained, and clinical assessments carried out, during an average 12 mo follow-up period. Yearly increments in height and weight were compared by HIV status, incidence of diarrhea and respiratory infections, and levels of sociodemographic variables. RESULTS: After adjusting for maternal education, anemia and vitamin A supplementation, HIV infection was related to 2.8 cm [95% confidence interval (95% CI) 0.6, 5.0] and 1.3 kg (95% CI 0.0, 2.5) lower yearly length and weight gains, respectively, in children who were between 6 and 11 mo old at baseline. Among children who were 12 23 mo old at recruitment, HIV infection was associated with 0.6 kg (95% CI 0.1, 1.0) less yearly weight gain. HIV infection was not related to linear or ponderal growth in children >24 mo old. Maternal illiteracy, severe child anemia and episodes of acute diarrhea were additional risk factors for growth delay in length. CONCLUSION: HIV infection is associated with linear and ponderal growth retardation in children aged <24 mo. Additional predictors of linear growth retardation include preventable conditions such as poor maternal education, child anemia and diarrheal disease. PMID- 15124843 TI - Cross-sectional study of allergic disorders in relation to familial factors in Japanese adolescents. AB - AIM: The roles of heredity and the household environment in the development of allergic disorders are not clearly established. This study examined the relationship between selected familial factors and the prevalence of symptoms of wheeze, atopic eczema and rhinoconjunctivitis in Japanese adolescents. METHODS: Study subjects were 5539 students aged 12-15 y in Suita City. A questionnaire ascertained gender, grade, number of older siblings, maternal age at childbirth, smoking in the household, domestic pets and parental history of allergy, as well as signs and symptoms of allergy in the previous 12 mo. The latter were based on diagnostic criteria from the International Study of Asthma and Allergies in Childhood. RESULTS: Male gender was independently associated with an increased prevalence of wheeze and a decreased prevalence of atopic eczema. The prevalence of rhinoconjunctivitis, but not wheeze or atopic dermatitis, significantly increased with advancing grade. A significant inverse dose-response relationship between the number of older siblings and the prevalence of rhinoconjunctivitis, but not wheeze or atopic dermatitis, was observed. Maternal age at childbirth, smoking in the household and domestic pets were not apparently related to any of the allergic disorders. A positive maternal allergic history was more evidently associated with an increased prevalence of wheeze and rhinoconjunctivitis, but not atopic eczema, than a positive paternal allergic history. CONCLUSION: The findings suggest that the factors associated with allergic disorders in Japan are largely the same as those already identified in Western populations. PMID- 15124844 TI - Urinary and faecal incontinence: a population-based study. AB - AIM: To investigate the coexistence of urine and faecal incontinence in Swedish schoolchildren. METHODS: Cohort study of all schoolchildren in the first and fourth grades in the city of Eskilstuna. A questionnaire was used, in which parents reported the prevalence of urine and faecal incontinence for their children. with a response rate of 67%. Data were analysed with multiple logistic regression. RESULTS: Daytime urinary incontinence (at least once a month) was reported in 6.3% of the first graders and 4.3% of the fourth graders, while bedwetting (at least once a month) was reported in 7.1% and 2.7% and faecal incontinence in 9.8% and 5.6%, respectively. Daytime urinary incontinence was strongly associated with faecal incontinence; adjusted odds ratio (OR) 7.2 (p < 0.001) as well as with bedwetting; OR 4.1 (p < 0.001), whereas faecal incontinence and bedwetting lacked a significant association (OR 1.2). CONCLUSION: This study demonstrates that soiling and daytime urinary incontinence often coexist in Swedish schoolchildren. Collaborative treatment strategies with gastroenterological and urological content need to be developed for these children. PMID- 15124845 TI - Serosurveillance for Japanese encephalitis in children in several districts of West Bengal, India. AB - AIM: To evaluate the prevalence of antibodies to flaviviruses, particularly Japanese encephalitis (JE) in children. METHODS: Virological and serological investigations were conducted into JE, along with dengue 2 (Den2) and West Nile viruses, in the Flavivirus group. The paediatric age group (up to 10 y), in the districts of Burdwan, Bankura, Midnapore and Purulia of West Bengal, India, was assessed for recent activity of these viruses, for 4 consecutive years from 1996 to 1999. In total, 2260 sera samples were collected, of which 204 were from acute fever cases; only 72 paired sera were available. There was a significantly higher incidence of fever cases in children belonging to the schedule caste and schedule tribes. These communities occupy the lowest level in the society and are closely associated with pigs. RESULTS: No virus could be isolated from the sera collected from fever cases. The results of the serological survey showed the presence of antibodies to JE virus in only 13.3% of the contact sera, 33.3% of acute sera and 22.1% of convalescent sera investigated. Only 3.1% of the contact sera had antibodies to Den2 and no antibodies were found against West Nile virus. This indicates that the JE virus is in recent circulation in the population of these four districts, and as the majority of children lack substantial immunity to JE virus, they may be affected by an epidemic. CONCLUSION: Children up to 10 y of age in the four districts of West Bengal are mostly non-immune to JE virus. If an outbreak occurs, the majority of these populations may be affected. PMID- 15124846 TI - Oesophageal atresia in premature infants: an analysis of morbidity and mortality over a period of 20 years. AB - AIM: To determine the morbidity and mortality of premature infants born with oesophageal atresia (OA) and to evaluate historical changes in morbidity and mortality over time. METHODS: Retrospective analysis of morbidity and mortality of all patients admitted for OA, with or without tracheo-oesophageal fistula, between 1982 and 2002. RESULTS: The study group consisted of 197 consecutive patients, of whom 55 (28%) were premature and 21 (11%) very premature. Type A atresia was found more often in very premature and premature infants than in those born at term (p = 0.02). Type E atresia was not found in the premature group (p = 0.004). At least one associated congenital anomaly was also present in 121 patients (61%). Postoperative complications developed more often in very premature and premature infants than in those born at term (p < 0.001). Gastro oesophageal reflux was diagnosed in 32/76 premature infants and in 41/121 term infants (p = 0.001). Mortality among very premature and premature infants was higher than among those born at term (p = 0.003). Withdrawal of treatment was the most frequent cause of death. CONCLUSION: Premature infants with OA have a higher morbidity and mortality than term infants with OA. The complications of prematurity contribute significantly to morbidity and mortality in premature infants with OA. There is no reason to refrain from the standard treatment of OA in premature infants with no severe associated congenital anomalies. PMID- 15124847 TI - Physical activity level and body mass index among schoolchildren in south-eastern Sweden. AB - AIM: The aims of this study were to identify current levels of physical activity and to study the relationship between physical activity and body mass index (BMI) using international cut-off points. METHODS: 871 children, aged 7-14 y, were measured for height and weight and the activity levels were analysed using pedometers to measure mean step counts for 4 consecutive days. RESULTS: Step counts were significantly higher in boys than in girls, and showed stability over age and large in-group differences. BMI showed that 13.2% of the boys and 14.5% of the girls were overweight and 4.5% of both boys and girls were obese. Analysis of step counts and BMIs for boys and girls revealed no significant correlations in any age group. CONCLUSION: Pedometers differentiate among age groups and gender concerning physical activity and facilitate individual goal-setting. The result of this study provide baseline information, useful as reference data, on youth physical activity as daily step counts, as well as on youth BMI levels according to new international cut-off points. PMID- 15124848 TI - Illness and exposure to negative life experiences in adolescence: two sides of the same coin? A study of 15-year-olds in Oslo, Norway. AB - AIM: To investigate associations between negative life experiences and common illnesses among adolescents. METHODS: Cross-sectional questionnaire study carried out at all lower secondary schools (10 grade) in Oslo. Norway, during 2000 and 2001 (n = 8316 pupils). Different negative life experiences and illnesses were addressed. RESULTS: The participation rate was 88%. Among reported negative life experiences last year were a pressure felt to succeed (62%), death of a close person (26%), exposure to physical violence (22%), bullying at school (15%) and sexual violation (4%). A large number of the pupils had some chronic illness: hay fever (38%), eczema (29%) and asthma (13%). Reported illnesses the previous 12 month were: headache (56%), painful neck or shoulders (35%), sore throat at least three times (15%), lower respiratory tract infection (9%) and mental problems for which help was sought (7%). During the week prior to the survey, 26% of all girls had symptoms of a depressive disorder, while this applied to 10% of all boys. Fifty-three percent of the boys (29% of the girls) who had depressive symptoms had been exposed to physical violence. Sexually violated boys had a high probability for seeking help for mental problems (OR = 4.9) and for frequent episodes of sore throat (OR = 2.5). Corresponding odds ratios for girls were 1.7 and 2.5, respectively. CONCLUSION: Common illnesses in adolescence are significantly associated with negative life experiences. In clinical encounters with adolescents not only should the presenting complaints be addressed, but also other common illnesses and relevant background factors such as negative life events. PMID- 15124849 TI - Immunogenicity and safety of a virosomal hepatitis A vaccine (Epaxal) in healthy toddlers and children in Chile. AB - In this open study, 20 toddlers and 80 schoolchildren received an intramuscular dose of Epaxal and a booster dose 12 mo later to assess the efficacy and safety of this aluminium-free, virosomal hepatitis A vaccine. Four weeks after primary vaccination, 94% of toddlers and 99% of schoolchildren had seroconverted, and all toddlers and 94% of schoolchildren remained seroprotected for 12 mo. CONCLUSION: After vaccination with Epaxal and booster, all subjects were seroprotected. Epaxal was very well tolerated by both age groups. PMID- 15124850 TI - Follow-up of acute pulmonary complications in cystic fibrosis by magnetic resonance imaging: a pilot study. AB - The aim of this pilot study was to obtain information on the value of MRI in the follow-up of atelectasis and pneumonic infiltrates in cystic fibrosis (CF). Six patients aged 5-15 y were initially examined using chest X-ray and magnetic resonance imaging (MRI). Both methods provided identical information. During follow-up, MRI proved suitable to monitor pulmonary complications. CONCLUSION: MRI of the lung is feasible and valuable in the follow-up of atelectasis and pulmonary infiltrates in CF. PMID- 15124851 TI - Historical views on celiac disease: the contribution of Adolf Baginsky (1843 1918). PMID- 15124852 TI - Barth syndrome without 3-methylglutaconic aciduria. AB - Barth syndrome involves cardiomyopathy, skeletal myopathy, neutropenia and 3 methylglutaconic (3-mgc) aciduria. 3-mgc aciduria has been observed in almost all reported cases and has served as a diagnostic criterion. CONCLUSION: A case of confirmed BTHS, but without 3-mgc aciduria, emphasizes the importance of extensive investigations in cases with suspected hereditary cardiomyopathy. PMID- 15124853 TI - Megestrol acetate promotes euglycemia and appetite in a child with persistent hyperinsulinemic hypoglycemia of infancy. AB - This report describes a successful treatment with megestrol acetate in a child with persistent hyperinsulinemic hypoglycemia of infancy (PHHI). An 8-y-old child with PHHI treated with octreotide had marked impairment of appetite sensation and feeding skills. Within 3 wk of starting megestrol acetate (8 mg/kg/d) to stimulate her appetite, she had a significant improvement. By 1 y postinitiation, she had acquired age-appropriate eating habits. The megestrol acetate caused hyperglycemia, necessitating the discontinuation of all other therapy for her hypoglycemia. Her height growth remained normal but she was found to have asymptomatic adrenal suppression. CONCLUSION: Megestrol acetate appeared to stimulate appetite and regulate glucose homeostasis in this child with PHHI. Additional studies will be required to document its efficacy and safety in other children with this disorder. PMID- 15124854 TI - Lymphadenitis cervicalis due to Mycobacterium interjectum in immunocompetent children. AB - This report presents two cases of cervical lymphadenitis due to Mycobacterium interjectum in healthy young children, identified by sequencing of the 16S rRNA gene. Surgical resection combined with chemotherapy resulted in cure. CONCLUSION: The attention of clinicians needs to be drawn to an emerging mycobacterial pathogen which might be overlooked or misidentified in routine laboratory testing. PMID- 15124855 TI - Congenital self-healing reticulohistiocytosis: report of a case and review of the literature. AB - Congenital self-healing reticulohistiocytosis (CSHRH), a rare disorder initially seen at birth or in the newborn period, is characterized by spontaneously involuting skin lesions. We present the case of a 12-d-old female who was born with many red-brownish nodules scattered over her entire body, and which regressed spontaneously. CONCLUSION: Although there are no clues to differentiate CSHRH from Letterer-Siwe disease, some signs and symptoms might enable us to reach an accurate diagnosis. A long-term follow-up to detect evidence of relapse or progression of the disease would be well advised. PMID- 15124856 TI - Impact of polymorphisms of beta2- and beta3-adrenergic receptor genes on longitudinal changes in obesity in early childhood. PMID- 15124857 TI - Dopexamine and cellular immune functions during systemic inflammation. AB - An immune-neuroendocrine interaction that is mediated via beta2-adrenergic receptors has been demonstrated previously. Dopexamine is a substance with strong beta2-adrenergic effects and is used in the treatment of critically ill patients. We therefore investigated the effect of dopexamine infusion on survival and cellular immune functions during systemic inflammation. Sepsis (CLP) was induced in male NMRI mice that received either 0.9% saline, dopexamine (0.05 mg/kg/hour ip, DPX), the selective beta2-adrenergic antagonist ICI 118.551 (0.5 mg/kg ip every 12 hours, ICI) or a combination of both drugs. 48 hours after onset of sepsis, survival rate, splenocyte apoptosis, splenocyte proliferation, splenocyte IL-2, IL-6 and IFN-gamma release, and leukocyte distribution were monitored. Dopexamine increased splenocyte apoptosis and normalized the distribution of circulating lymphocytes but did not affect sepsis-induced mortality. ICI 118.551 induced a dramatic increase of mortality paralleled by a decreased splenocyte proliferation and the strongest increase in splenocyte apoptosis. Co administration of dopexamine abolished the ICI 118.551-induced alterations but this effect seemed to be mediated via a pathway other than adrenergic beta2 receptors. We conclude that dopexamine modulates cellular immune functions during systemic inflammation and that different receptor systems are involved in the mediation of this process. Furthermore, the immunomodulatory effect of beta2 adrenergic blockade was demonstrated. PMID- 15124858 TI - Ubiquitin-like polypeptide inhibits cAMP-induced p38 MAPK activation in Th2 cells. AB - Ubi-L, an isoform of the monoclonal nonspecific suppressor factor (MNSF), is an 8.5-kDa ubiquitin-like polypeptide. Ubi-L exhibits an antigen-nonspecific immunosuppressive function on various target cells including murine T helper type 2 (Th2) clone, D10 cells. Ubi-L specifically binds to cell surface receptors on D10 cells. In this study, we observed that Ubi-L inhibited cAMP-induced IL-5 mRNA expression in D10 cells but not in thymoma cell line EL4. In addition, Ubi-L effectively inhibited cAMP-induced p38 MAPK activation in D10 cells. Ubi-L also showed inhibitory activity on IL-5 and IL-13 production by D10 cells stimulated with phorbol ester plus dibutyryl cAMP. Furthermore, Ubi-L inhibited IL-4 production in Th2 cells derived from primary CD4+ T cells. PMID- 15124859 TI - Bacterial lipopolysaccharide induces proliferation of IL-6-dependent plasmacytoma cells by MAPK pathway activation. AB - Mouse plasmacytomas are appropriate models to study the biology of human multiple myeloma (MM). Growth of murine interleukin-6 (IL-6)-dependent hybridoma/plasmacytoma lines can be stimulated by bacterial lipopolysaccharides (LPS). However, the molecular mechanisms of this phenomenon are still not elucidated. In this study the in vitro action of bacterial LPS on the mouse IL-6 dependent B9 hybridoma/plasmacytoma cell line and two IL-6-dependent hybridomas was investigated. The involvement of different signal transduction pathways was established using specific kinase inhibitors in proliferation assays and immunoblotting analysis of the kinase activity. Selective mitogen-activated protein kinase (MAPK) kinase inhibitor PD989059 inhibited both IL-6- and LPS induced B9 cell proliferation. In contrast, in H187 and H188 cells, PD98059 inhibited only LPS-, but not IL-6-stimulated cell growth. The kinetics of MAPK activation in all cell lines showed that phosphorylation of p42 MAPK (encoded as ERK2) but not of p44 MAPK (ERK1), was considerably increased after treatment with LPS. We found that in H187 and H188 hybridomas IL-6 induced proliferation by a different STAT3-dependent mechanism. This study demonstrates the key role of the MAPK pathway in LPS-stimulated growth of mouse IL-6-dependent plasmacytoma cells. These findings suggest the presence of signaling mechanism in MM cells inducible by bacterial mitogens and possibly mediated by Toll-like receptors (TLR)- evolutionarily conserved molecules playing a central role in the microbial recognition and initiation of the cellular innate immune response. PMID- 15124860 TI - Lymphocyte subpopulations from patients with primary antibody deficiency do not show increased telomere erosion. AB - Telomere erosion and residual replicative capacity can be used as markers of the replicative history of somatic cells. We have investigated telomere length, in vitro replicative capacity and rate of telomere erosion in T and B lymphocyte populations from patients with primary antibody deficiency requiring immunoglobulin replacement therapy. We found no significant differences in telomere lengths of B cells, or of CD4+, CD8+, CD45RA+ (naive) and CD45RO+ (memory) T cell populations between patients and age matched controls. Overall, telomere length correlated inversely with age, and was reduced in memory (CD45RO+) as compared with naive (CD45RA+) T cells. In vitro long-term (6 months) cell cultures showed no differences between patients and controls in the mitogen stimulated replicative potential of T cell subpopulations (CD4+, CD8+, CD45RA+, CD45RO+), or in the rates of telomere erosion with cellular replication in these cell populations. The rate of telomere erosion per population doubling in CD45RA+ cells, however, was greater than in CD45RO+ cells in both patients and controls. These data suggest that premature immune exhaustion is unlikely to represent a long-term complication of primary antibody deficiency. PMID- 15124861 TI - Elimination of activated but not resting primary human CD4+ and CD8+ T cells by Fas ligand (FasL/CD95L)-expressing Killer-dendritic cells. AB - Dendritic cells (DC) genetically engineered to express high levels of Fas ligand (FasL/CD95L) have been demonstrated to delete T cells in an antigen specific manner in several different animal models in vivo. However, the immunomodulatory capacity of primary human FasL-expressing Killer-DC has not been determined. Therefore, human Killer-DC were generated from mature monocyte-derived DC using the inducible CRE/LoxP adenoviral vector system, and the immunoregulatory capacity of these cells was analyzed in cocultures with primary human T cells in vitro. Combined transductions of DC by AdloxPFasL and AxCANCre resulted in FasL expression in > 70% of DC without affecting the mature phenotype. Proliferation of activated primary human T cells was inhibited up to 80% in cocultures with FasL-expressing DC but not EGFP-transduced DC, which was due to induction of apoptosis in activated but not resting CD4+ and CD8+ T cells. Apoptosis induced by Killer-DC could be blocked by an anti-FasL-antibody in a dose dependent fashion. The present results demonstrate that FasL-expressing Killer-DC eliminate activated but not resting primary human CD4+ and CD8+ T cells by induction of Fas mediated apoptosis supporting the concept to apply Killer-DC as a novel strategy for the treatment of T cell-dependent autoimmune disease and allograft rejection in humans. PMID- 15124862 TI - Protothecosis. AB - Protothecosis is an infection caused by achlorophyllic algae of the genus Prototheca which rarely affects humans. Some 100 cases have been described in the medical literature, the majority caused by the species P. wickerhamii. The skin is the organ most frequently involved. Diagnosis is performed by isolation of the microorganism in culture or by histopathology. The ideal treatment has not been defined, with amphotericin B and the azoles having been employed. Surgical excision is recommended for small, localized lesions. We describe a case of cutaneous protothecosis on the right fourth finger of a female patient 59 years old with no underlying disease. Administration of itraconazole 400 mg/day for 6 weeks failed to produce an adequate clinical response. Treatment was then changed to fluconazole 200 mg/day, with regression of the lesion. PMID- 15124863 TI - Pathogenesis of indoor fungal diseases. AB - Mold growth within homes and other buildings has been associated to varying degrees with human health problems. These problems vary from allergenic disease to toxicosis. Case definitions for mold exposure have not been adequately defined to allow for a pathognomonic diagnosis of mold-caused disease following indoor exposure. Some important factors that may contribute to the pathogenesis of indoor mold induced disease include beta (1,3)-D-glucans, outer cell wall fungal hydrophobins, 1,8-dihydroxynaphthalene melanin, fungal volatile organic compounds, mycotoxins, and stachylysin. The information in this contribution was presented as the ISHAM Presidential address as a means to clarify some of the confusing surrounding indoor mold-related health issues. PMID- 15124864 TI - Differential expression of actin modulated by temperature in mycelial and yeast cells of Paracoccidioides brasiliensis. AB - We used yeast and mycelial forms of Paracoccidioides brasiliensis to evaluate the effect of heat shock stress on actin expression. P. brasiliensis yeasts harvested during the exponential growth phase showed more expression of the actin mRNA when incubated at 40 degrees C than when incubated at 37 degrees C, the usual temperature at which these yeasts grow. In contrast, expression of actin mRNA was lower in yeasts incubated at 25 degrees C than in yeasts incubated at 37 degrees C. Mycelium harvested at 25 degrees C, an approximation of its normal growth temperature, and then exposed to 37 degrees C and 40 degrees C showed progressively higher expression of actin mRNA. Mycelial and yeast forms showed a similar pattern of response to exposure to supra-optimal temperatures: both showed the same increase in expression of actin. This suggests that actin may play a role not only in cellular differentiation but also in this species' rapid adaptive response to heat stress, a mechanism necessary to deal with a potentially hostile environment. PMID- 15124865 TI - Cytokines in mice treated with amphotericin B-intralipid. AB - Amphotericin B (AMB) intralipid (IL) admixtures (AMB-IL) are composed of components approved for clinical use and are commercially available at low cost. They are stable and exhibit in-vitro and in-vivo efficacy against Candida infections, as well as resulting in significantly reduced toxicity in comparison with that of conventionally administered amphotericin B. We examined the production of cytokines in uninfected mice treated with AMB or AMB-IL, as evaluated by expression of mRNA corresponding to the cytokines. Expression was measured by intensity of bands in comparison to the intensity of beta-actin control bands, with the latter assigned an arbitrary standard value of 100% and other bands measured in relative percentages. We found that both in naive and compromised mice, AMB treatment caused significantly greater production of the pro-inflammatory cytokines tumour necrosis factor alpha (TNF-alpha) and interleukin 1 beta (IL-1beta) than was seen in animals treated with AMB-IL or with another lipid AMB formulation, AmBisome. We hypothesize that the superior tolerance for the AMB-IL admixtures, as compared with conventional AMB, might derive from the reduced expression of the pro-inflammatory cytokines. TNF-alpha and IL-1beta, which mediate many potentially adverse pathophysiological events similar to those seen as side-effects of AMB usage. PMID- 15124866 TI - Subcutaneous phaeohyphomycotic abscess caused by Pleurophomopsis lignicola. AB - A 41-year-old man with a past medical history of diabetes mellitus type II, AIDS (diagnosed 2 years earlier; CD4 count < 10), peripheral neuropathy, and pulmonary tuberculosis of 2 years duration was admitted to the hospital with abnormal liver function tests. There was a chronic hepatitis/cholestasis that had worsened while the patient was undergoing directly observed tuberculosis therapy. On admission, the patient complained of a painful swelling on his right arm. In the posterior aspect of the arm, there was a 3-4-cm subcutaneous mass that was fluctuant, mobile and tender. Incision of the mass released yellowish pus mixed with blood. Direct examination of the pus in KOH mounts and Gram-stained smears revealed subhyaline, septate, branched hyphae. When the pus was cultured on Sabouraud dextrose agar containing chloramphenicol, several velvety, olivaceous grey colonies grew after 7 days at 25 degrees C. When grown on oatmeal agar, the fungus produced subglobose, rostrate pycnidia with phialidic conidiogenous cells, and 1-celled cylindrical conidia. It was identified as Pleurophomopsis lignicola Petrak. This report describes the third known case of subcutaneous infection caused by P. lignicola in an immunocompromized patient. PMID- 15124867 TI - Molecular typing of Cryptococcus neoformans by PCR fingerprinting, in comparison with serotyping and Fourier transform infrared-spectroscopy-based phenotyping. AB - Molecular typing by PCR fingerprinting using the single primer (GACA)4 was performed with 110 isolates of Cryptococcus neoformans. Seventy clinical isolates of C. neoformans var. neoformans from Germany (n = 52) and Africa (n = 18) were included. Of these, serotype A (C. neoformans var. grubii) accounted for 47 isolates, serotype D for 12 and serotype AD for 11. Fourier transform infrared (FT-IR) spectroscopy was evaluated for its discriminatory power in phenotyping. Molecular types, defined by different PCR fingerprinting patterns, were compared to serotypes, and both sets of results were compared with the results of analysis by FT-IR spectroscopy. PCR fingerprinting revealed genotypic diversity within each serotype; it showed three different genotypes (designated VNA1-VNA3) within serotype A, two within serotype D (VND1 and VND2), and three within serotype AD (VNAD1-VNAD3). The nomenclature of molecular types within C. n. var. neoformans, as seen in publications to date, is not uniform. In this study, the name assigned to each genotype was based on the 98.6% concordance of genotypes with serotypes, a correspondence that facilitates interlaboratory comparison. This nomenclature is tentatively recommended as a standard. FT-IR spectroscopy combined with hierarchical cluster analysis successfully distinguished C n. var. neoformans from C. n. var. gattii. For C. n. var. neoformans, FT-IR confirmed three distinct genotypes within serotype A and was able to distinguish isolates derived from particular patients as well as isolates differing at the sub-genotype level. Within C. n. var. gattii, the serotypes B and C did not correlate with the four genotypes VGI-VGIV. However, these serotypes could clearly be separated by FT-IR spectroscopy. The molecular profiles were reproducible, and were more stable and more discriminating than serotyping. In connection with a standardized nomenclature, PCR fingerprinting can be a beneficial tool for global epidemiological studies. FT-IR spectroscopy adds an additional level of resolution. PMID- 15124868 TI - Cryptococcus neoformans var. gattii can exploit Acanthamoeba castellanii for growth. AB - It has recently been proposed that the origin and maintenance of virulence in certain environmental fungi is influenced by their interactions with non vertebrate hosts such as amoebae and nematodes. In prior studies we have shown that the interactions of the soil amoebae Acanthamoeba castellanii with Cryptococcus neoformans varieties neoformans and grubii resemble those with macrophages. Here we extend those studies to C. neoformans variety gattii and describe quantitative differences in the type and outcome of the interactions observed relative to the other varieties. C. neoformans var. gattii proliferated in the presence of A. castellanii but the interaction was primarily extracellular with a paucity of phagocytic events. Experiments with acapsular cells coated with polysaccharide suggest that differences in the capsule structure may be responsible for the different interactions between cells of varieties neoformans, grubii, and gattii with amoebae. The ability of C. neoformans var. gattii to exploit amoebae indicates that despite major biological differences between C. neoformans varieties, all retain the ability to be pathogenic for A. castellanii. PMID- 15124869 TI - Antifungal drug response in an in vitro model of dermatophyte nail infection. AB - Despite terbinafine being fungicidal against Trichophyton rubrum in standard NCCLS assays and rapidly accumulating in nails in vivo, onychomycosis patients require prolonged terbinafine treatment to be cured. To investigate this, we developed a more clinically relevant onychomycosis in vitro test model. Human nail powder inoculated with T. rubrum and incubated in liquid RPMI 1640 salt medium, which did not support growth alone, developed extensive and invasive mycelial growth. Antifungal drugs were added at different concentrations and cultures incubated for 1 to 4 weeks. Fungal survival was determined by spreading cultures on PDA plates without drug and measuring CFU after 1 to 4 weeks incubation. Drug activity was expressed as the nail minimum fungicidal concentration (Nail-MFC) required for 99.9% elimination of viable fungus. Terbinafine Nail-MFC was 4 microg/ml after 1 week exposure, decreasing to 1 microg/ml after 4 weeks exposure, much higher than MFCs < or = 0.03 microg/ml determined in standard NCCLS MIC assays. In contrast, other clinically used drugs were unable to kill T. rubrum after 4 weeks incubation in this model. Invasive mycelial growth on nail appears to protect T. rubrum from the cidal action of systemic drugs, thus providing a rationale for the long treatment periods in onychomycosis. PMID- 15124870 TI - Clinical, histopathological and immunological effects of exposure of canine skin to Malassezia pachydermatis. AB - The effects of the daily application for 7 days of suspensions of Malassezia pachydermatis to normal canine skin were evaluated in 10 beagle dogs. Four out of six dogs challenged without occlusion developed transient lesions generally characterized clinically by mild erythema with papules and histologically by mild epidermal hyperplasia and a superficial perivascular dermatitis. Saline-treated control sites showed no clinical signs. In four dogs challenged with occlusion, skin lesions occurred at both yeast and saline-treated sites; erythema and papules were more severe at the yeast-treated sites in three dogs. Occlusion induced more persistent lesions, which resolved within 24 days. Population densities of the yeast were highest at day 8 and declined rapidly following cessation of application. Peripheral blood mononuclear cell proliferation indices following M. pachydermatis exposure in vitro and serum concentrations of M. pachydermatis-specific IgG antibodies did not vary significantly during the study. Delayed (24 h) intradermal test reactivity to M. pachydermatis antigens developed in all eight dogs with clinical signs following yeast exposure. This study suggests that the resistance of healthy canine skin to infection by M. pachydermatis is mediated by local delayed hypersensitivity responses and, or innate epidermal immune mechanisms. PMID- 15124871 TI - Zygomycosis caused by Cunninghamella bertholletiae in a kidney transplant recipient. AB - Infections caused by Cunninghamella bertholletiae are rare but severe. Only 32 cases have been reported as yet, but in 26 of these this species was a contributing cause of the death of the patient. This opportunistic mould in the order Mucorales infects immunocompromized patients suffering from haematological malignancies or diabetes mellitus, as well as solid organ transplant patients. The lung is the organ most often involved. Two cases of primary cutaneous infection have been previously reported subsequent to soft-tissue injuries. We report a case of primary cutaneous C. bertholletiae zygomycosis in a 54-year-old, insulin-dependent diabetic man who was treated with tacrolimus and steroids after kidney transplantation. No extracutaneous involvement was found. In this patient, the infection may have been related to insulin injections. The patient recovered after an early surgical excision of the lesion and daily administration of itraconazole for 2 months. This case emphasizes the importance of an early diagnosis of cutaneous zygomycosis, which often presents as necrotic-looking lesions. Prompt institution of antifungal therapy and rapid surgical intervention are necessary to improve the prospects of patients who have contracted these potentially severe infections. PMID- 15124872 TI - Serotype distribution of Cryptococcus neoformans in patients in a tertiary care center in India. AB - The prevalence of specific serotypes of Cryptococcus neoformans in a given area bears on regional epidemiological patterns, the expected spectrum of clinical disease, and predicted response to therapy. In this retrospective study we analyzed the serotypes of 45 degrees C neoformans isolates from 36 North Indian patients with varied clinical presentations. The majority of the isolates were serotype A (87%), and surprisingly, a significant number were serotype B (five isolates, 11%), which caused infection in patients both positive and negative for HIV. One unusual isolate was not typable with factor sera. Study of serotype distribution in patients showed serotypes A and B to be present, respectively, in 92% and 8% of 36 patients. In one apparently immunocompetent patient two serotypes, A and B, were isolated simultaneously from two different sites, lung and scalp abscess. This is the first reported case in which an individual was infected with two serotypes at the same time. In one HIV-infected child serotype A was isolated from blood. Our results suggest that the distribution of serotypes in Indian clinical isolates is different than that found in other regions. PMID- 15124873 TI - USDA plans sharp increase in BSE testing. PMID- 15124874 TI - Town hall meeting surveys diversity in the profession. PMID- 15124875 TI - PRRS: is elimination attainable? PMID- 15124876 TI - Shelter medicine: a budding field that is helping to raise the standard of care in animal shelters. PMID- 15124877 TI - Questions about dental radiograph. PMID- 15124878 TI - Opposes pet insurance. PMID- 15124879 TI - Educating the next generation of swine veterinarians: needs and strategies. AB - The education of future swine veterinarians will require a team approach from academic institutions, private practitioners, and the pork industry Students will need to be active participants in their education. University and college administration, faculty, industry, government, professional organizations, and private practitioners will all need to provide the leadership and financial framework to support the future of veterinary education related to swine. PMID- 15124880 TI - What is your diagnosis? Fracture of the fibula. PMID- 15124881 TI - Animal behavior case of the month. A cockatiel was examined because of repetitive chewing of the third digit of the right foot. PMID- 15124882 TI - The informed consent doctrine: what veterinarians should tell their clients. PMID- 15124883 TI - Outbreaks of zoonotic enteric disease associated with animal exhibits. PMID- 15124884 TI - Interactions between lipopolysaccharide and the intestinal epithelium. PMID- 15124885 TI - Evaluation of the effect of cephalexin and enrofloxacin on clinical laboratory measurements of urine glucose in dogs. AB - OBJECTIVE: To determine the effects of cephalexin and enrofloxacin on results of 4 commercially available urine glucose tests in dogs. ANIMALS: 6 healthy adult female dogs. PROCEDURE: In a crossover design, cephalexin (22 and 44 mg/kg [10 and 20 mg/lb], p.o., q 8 h) or enrofloxacin (5 and 10 mg/kg [2.3 and 4.5 mg/lb], p.o., q 12 h) was administered to dogs for 1 day. Urine samples were tested for glucose at 0, 6, and 24 hours after drug administration. In vitro, dextrose was added to pooled glucose-negative canine urine samples containing either no antimicrobial or known concentrations of either antimicrobial; urine samples were then tested for glucose. RESULTS: In vivo, false-positive results were obtained by use of a tablet test in the presence of both antimicrobials and by use of a strip test in the presence of cephalexin. In vitro, false-positive results were obtained with the tablet test at the highest urine concentration of cephalexin (2,400 microg/mL) and with a strip test at the highest concentration of enrofloxacin (600 microg/mL). Enrofloxacin in urine samples containing dextrose caused the urine glucose tests to underestimate urine glucose concentration. CONCLUSIONS AND CLINICAL RELEVANCE: Cephalexin and enrofloxacin at dosages used in clinical practice may result in false-positive or false-negative urine glucose results, and care should be taken when using urine as a basis for identifying or monitoring diabetic animals. PMID- 15124886 TI - Prevalence and characteristics of pain in dogs and cats examined as outpatients at a veterinary teaching hospital. AB - OBJECTIVE: To determine prevalence of pain among dogs and cats examined as outpatients at a veterinary teaching hospital and characteristics of pain in dogs and cats with evidence of pain. DESIGN: Cross-sectional study. ANIMALS: 1,153 dogs and 652 cats examined as outpatients at The Ohio State University during 2002. PROCEDURE: A questionnaire was administered to owners of all dogs and cats. For dogs and cats with evidence of pain, the cause, signs, anatomic location, type (superficial somatic, deep somatic, or visceral), duration, and severity of the pain and the principle mechanism (inflammatory, neuropathic, both, or unknown) responsible for the pain were determined on the basis of questionnaire responses and results of physical examination. The presence of primary hyperalgesia, secondary hyperalgesia, allodynia, and hyposensitivity was recorded. RESULTS: 231 (20%) dogs and 92 (14%) cats had evidence of pain. Dogs with evidence of pain were significantly older and heavier than dogs without. Cats with evidence of pain were significantly older than cats without. In most dogs and cats with evidence of pain, the pain was determined to be of short duration (< 7 days), of mild or moderate severity, somatic, associated with primary hyperalgesia, and inflammatory. Analgesic drugs were frequently administered to dogs with chronic pain, but were not always considered effective. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that mild or moderate pain associated with inflammation may be seen in dogs and cats examined as outpatients. Older, heavier dogs and older cats were more likely to have evidence of pain. PMID- 15124887 TI - Cardiogenic hypertrophic osteopathy in a dog with a right-to-left shunting patent ductus arteriosus. AB - A 5-year-old castrated male Shetland Sheepdog was examined because of progressive bilateral hind limb thickening. Cyanosis of the preputial mucous membranes was evident, whereas the oral mucous membranes had a normal color. A well-structured, palisade-like periosteal reaction with no underlying bone destruction was evident on radiographs of the hind limbs. The radiographic changes were consistent with hypertrophic osteopathy (HO). Severe right-sided cardiomegaly was seen on thoracic radiographs, and a diagnosis of patent ductus arteriosus with right-to left shunting was made by means of echocardiography and contrast echoaortography. The cyanotic heart disease was believed to be the cause of the HO. Hypertrophic osteopathy has been associated with a number of diseases in animals and humans. In humans, congenital heart defects that cause cyanosis are among the most common causes of HO. PMID- 15124888 TI - Ultrasonographic diagnosis of septic arthritis secondary to porcupine quill migration in a dog. AB - A 7-year-old castrated male German Shepherd Dog was evaluated for lethargy, icterus, and sepsis. Porcupine quills had been removed from the dog's face 1 month prior to examination; progressive right forelimb lameness had developed soon after removal of the quills. Septic arthritis of the right elbow joint was diagnosed and was unresponsive to antimicrobial and anti-inflammatory treatments. At the time of referral, the dog had developed endocarditis, septicemia, and disseminated intravascular coagulation. Via ultrasonography, a foreign body consistent with a porcupine quill was detected in the medial portion of the right humeroradial joint. The dog did not respond to initial supportive treatment and died as a result of cardiac arrest. Postmortem examination confirmed the presence of a quill in the medial compartment of the right elbow joint and severe acute endocarditis with septic emboli to the kidneys and spleen. Ultrasonographic examination should be considered as a diagnostic tool when septic arthritis secondary to a foreign body is suspected in dogs. PMID- 15124889 TI - Retroperitoneal sarcomas in dogs: 14 cases (1992-2002). AB - OBJECTIVE: To describe the clinical features, surgical and histologic findings, biological behavior, and outcome of dogs with retroperitoneal sarcomas. DESIGN: Retrospective study. ANIMALS: 14 dogs. PROCEDURES: Medical and pathology records from 1992 to 2002 of dogs with tumors originating in the retroperitoneal space were reviewed. Dogs with retroperitoneal tumors originating from the adrenal glands, kidneys, or ureters were excluded. Inclusion criteria included observation of a tumor arising from the retroperitoneal space during exploratory surgery or necropsy and histologic confirmation of tumor type. Details of clinical signs, diagnostic findings, surgical management, and outcome were determined from medical records and telephone interviews with veterinarians and owners. RESULTS: Retroperitoneal sarcoma was diagnosed in 14 dogs, 2 at necropsy and 12 during exploratory surgery. Hemangiosarcoma was the most common histologic diagnosis. Seven dogs had regional extension of the sarcoma into adjacent organs, and 4 dogs had metastatic disease. Grossly complete resection was possible in 6 dogs. Cytoreductive surgery or incisional biopsy was performed in the remaining dogs. Two dogs were treated with palliative radiation therapy (1 intraoperatively and 1 postoperatively). Three dogs received adjunctive chemotherapy, although none completed the targeted course because of development of local recurrence or metastatic disease. Local recurrence was reported in 2 of 12 dogs and metastasis in 10 of 14 dogs. Thirteen dogs died or were euthanatized as a result of the retroperitoneal sarcoma; 1 dog was alive and disease-free 410 days after surgery. Median survival time was 37.5 days. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, retroperitoneal sarcomas are aggressive tumors with a high rate of local recurrence and metastasis, and a poor survival time. PMID- 15124890 TI - Surgical reduction and stabilization for repair of femoral capital physeal fractures in cats: 13 cases (1998-2002). AB - OBJECTIVE: To evaluate anatomic reduction and surgical stabilization of femoral capital physeal fractures in cats. DESIGN: Retrospective study. ANIMALS: 13 cats. PROCEDURE: Medical records of cats with unilateral or bilateral femoral capital physeal fractures evaluated from 1998 to 2002 were reviewed. Age and weight of cats at the time of surgery; breed; sex; concurrent injuries; severity of lameness before and 1, 2, 4, 6, and 8 weeks after surgery; the amount of fracture reduction achieved and number of Kirschner wires (K-wires) used; degree of degenerative joint disease of the hip joint and lysis of the femoral neck and head observed after surgery; whether K-wires were removed after surgery; and complications after surgery were evaluated. RESULTS: Thirteen cats with 16 capital physeal fractures were identified. There was significant improvement in the severity of clinical lameness in all cats from weeks 1 through 4 after surgery. There was no correlation between the scores of the individuals who evaluated radiographs for fracture reduction and placement of K-wires. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that surgical stabilization and repair of femoral capital physeal fractures facilitate a short recovery period and a good prognosis for return to normal function in cats. PMID- 15124891 TI - Radiotherapy of a recurrent ossifying fibroma in the paranasal sinuses of a horse. AB - A 7-year-old female Thoroughbred was admitted with a history of labored breathing, stridor, and exercise intolerance. Examination revealed a mass in the left paranasal sinuses that was determined to be an ossifying fibroma. Initial treatment consisted of surgical removal of the mass alone; however, the mass recurred 9 months after surgery. The mass was again removed, and adjunctive radiotherapy consisting of 3,000 cGy of cobalt radiation was administered. This time, the tumor did not recur for > 6 years. A third surgery was performed to remove the mass, and adjunctive radiotherapy consisting of 4,000 cGy of photon beam radiation from a linear accelerator was administered. The mass did not recur during the subsequent 3 years. Ossifying fibromas are uncommon tumors that frequently recur if incompletely excised. Results in this horse suggest that adjunctive radiotherapy may delay or prevent tumor recurrence in affected horses. PMID- 15124892 TI - Risk factors for umbilical hernia in Holstein heifers during the first two months after birth. AB - OBJECTIVE: To determine risk factors associated with identification of an umbilical hernia during the first 2 months after birth in Holstein heifers. DESIGN: Case-control study. ANIMALS: 322 Holstein heifers born in a single herd (45 with an umbilical hernia and 277 without). PROCEDURE: Risk factors that were examined included sire, whether the dam had a history of umbilical hernia, milk yield, duration of gestation, whether the dam had a history of dystocia, whether the heifer had a twin, birth weight, total serum protein concentration, and whether the heifer had an umbilical infection. Logistic regression was used to analyze risk factors. RESULTS: Heifers born to sires with > or = 3 progeny with an umbilical hernia were 2.31 times as likely to develop an umbilical hernia as were heifers born to sires with < or = 2 progeny with an umbilical hernia. Heifers with umbilical infection were 5.65 times as likely to develop an umbilical hernia as were heifers without umbilical infection. CONCLUSIONS AND CLINICAL RELEVANCE: Sire and umbilical infection were associated with risk of an umbilical hernia during the first 2 months of life in Holstein heifers. Attributable proportion analysis indicated that the frequency of umbilical hernias in Holstein heifers with umbilical infection would have been reduced by 82% if umbilical infection had been prevented. PMID- 15124893 TI - Significant prognostic factors for Vogt-Koyanagi-Harada disease in the early stage. AB - This study identified possible prognostic signs in the acute stage of Vogt Koyanagi-Harada (VKH) disease in a retrospective chart review of all patients diagnosed with VKH disease between 1991 and 2001. Those who were diagnosed more than 1 month after the onset of ocular symptoms were excluded. Data recorded included age, sex, clinical features, systemic manifestations, recurrence, treatment, complications, and final visual acuity. Exudative retinal detachment was ranked into 3 grades (grade 1: within peripapillary 3 disc diameters and arcade; grade 2: larger than grade 1 but no inferior retinal detachment; grade 3: inferior or total retinal detachment). Of the 31 patients, 19 were males and 12 were females. Mean age at presentation was 38.55 +/- 10.63 years. The mean follow up period was 33.09 months. Extraocular manifestations were present in 17 cases. Four patients had at least one complication, including cataract in seven eyes and glaucoma in one eye. Forty-nine eyes (79%) had a final visual acuity of 6/12 or better. Final visual acuity was significantly better in younger patients (p = 0.023) and those who had less extensive retinal detachment (p = 0.006), no pigmentary change (p = 0.008) and no complications (p = 0.030). The visual prognosis of VKH disease is variable, though generally favorable. In the acute stage, the extent of retinal detachment may be an important risk factor for visual outcome. Further study of this factor as an indicator for treatment is necessary. PMID- 15124894 TI - Physician practices in the diagnosis of carpal tunnel syndrome at a medical center in southern Taiwan. AB - Carpal tunnel syndrome (CTS) is the most common neurologic entrapment disorder diagnosed in the upper limb. Nevertheless, there is still debate about the most reliable test that should be performed to diagnose CTS. Much of the argument has been drawn from the opinions of individuals or groups with varying degrees of expertise in the field; little has been based on actual data. The purpose of this study was to investigate the diagnostic patterns of CTS in an academic medical setting in southern Taiwan. The charts of 1,050 patients with a diagnosis of CTS over a 1-year period (2001-2002) were retrospectively reviewed. Data on 622 patients with new-onset CTS were included in the analysis. On the patient's initial visit, physicians made a diagnosis of CTS in 34.9% of cases solely on the basis of the history of symptoms without resort to provocative tests, while 8.7% of cases were diagnosed on the basis of symptom characteristics alone in spite of negative provocative tests. A CTS diagnosis was given according to symptoms and positive provocative tests in 55% of cases. Apart from these, CTS diagnosis remained unchanged in 27.3% of cases without electrodiagnostic signs of CTS during follow-up visits. An average of 1.6 diagnostic maneuvers were conducted for CTS patients, with nerve conduction velocity (NCV) studies (516 cases) being the most frequently performed, followed by Tinel's sign (350 cases) and Phalen's test (102 cases). Our findings imply that physicians are inclined to base their diagnosis on clinical history and physical examination for patients with suspected CTS. Clear guidelines regarding the indications for referral for NCV studies should be established in response to the increased concerns about the cost effectiveness of diagnostic tests. PMID- 15124895 TI - Potential factors associated with contraceptive intention among adolescent males in Taiwan. AB - This study explored the predictors of contraceptive intention in adolescent males in Taiwan. It used a cross-sectional design and a sample of 1,000 vocational high school male students to determine the potential factors associated with contraceptive intention. Data were collected on an anonymous, self-administered questionnaire, including personal background variables, prior sexual experience, contraceptive knowledge, contraceptive attitude, contraceptive self-efficacy, perception of peers' contraceptive behavior, perception of support from significant others, and parental-adolescent communication about contraception. A total of 230 participants (23%) had prior sexual experience, 13% (30) of whom reported being involved in their girlfriends' pregnancies. Of the participants, 45% recognized the use of condoms as the most favorable contraceptive method for future sexual intercourse. Second to condom use, the combination of the calendar method and condom use was also popular (13.1%). Multiple stepwise regression indicated that better contraceptive attitudes, higher contraceptive self efficacy, no prior sexual experience, more perception of peers' contraceptive behavior, and higher perception of support from significant others were predictors of higher contraceptive intention. The above predicting factors explained 31.9% of the total variance for contraceptive intention among adolescents. These results provide health professionals with important information to understand the reality adolescents encounter and to design effective contraceptive programs for male adolescents. PMID- 15124896 TI - Serum concentration of soluble decoy receptor 3 in glioma patients before and after surgery. AB - The suppression of immune responses in malignant gliomas is thought to be involved in glioma pathogenesis. The newly identified tumor-secreted soluble decoy receptor 3 (DcR3) can bind to the ligands CD95L and LIGHT, thereby neutralizing their pro-apoptotic actions. Little is known of the production of DcR3 by glioma cells. This study investigated the serum concentration of DcR3 in glioma patients before and after tumor removal. Blood samples were taken from 17 glioma patients and 10 control patients. The serum DcR3 concentration was measured using a DcR3 enzyme-linked immunosorbent assay. There was no statistically significant difference between preoperative (0.069 +/- 0.027 ng/mL) and postoperative DcR3 concentrations (0.068 +/- 0.022 ng/mL; p = 0.951). Similarly, there was no difference in preoperative DcR3 concentration between glioma patients (0.069 +/- 0.027 ng/mL) and controls (0.063 +/- 0.023 ng/mL; p = 0.106). Our study demonstrated no alteration in DcR3 concentration in glioma patients before and after tumor removal. PMID- 15124897 TI - Pure red cell aplasia after ABO major-mismatched allogeneic peripheral blood stem cell transplantation successfully treated with plasma exchange and low-dose steroid: two case reports. AB - Pure red cell aplasia (PRCA) is a complication of ABO-incompatible allogeneic stem cell transplantation. The mechanism is not well known, although the isoagglutinin titer before transplantation or cyclosporine use is considered to be the cause. Patients with this complication require more blood transfusions than those without it. There is no standard treatment. We report two cases of PRCA after allogeneic peripheral blood stem cell transplantation that were successfully treated with plasma exchange and low-dose steroid. PMID- 15124898 TI - Multiple symmetric lipomatosis (Madelung's disease): report of two cases. AB - Multiple symmetric lipomatosis (Madelung's disease) is a rare disease characterized by non-encapsulated adipose deposits in the neck, the superior part of the trunk and, very rarely, in the limbs. It is common in middle-aged Caucasian Mediterranean males but very rare in Chinese people. We describe two cases of Madelung's disease: both patients presented complaining of a development of a painless neck mass that was increasing in size. Panendoscopy was normal. Both underwent incisional biopsies that revealed benign adipose tissue characterizing Madelung's disease. We discuss the clinical presentation, associated morbidity, and treatments for Madelung's disease. PMID- 15124899 TI - Unusual upper gastrointestinal bleeding due to late metastasis from renal cell carcinoma: a case report. AB - A case of recurrent massive upper gastrointestinal bleeding originating from metastatic renal cell carcinoma is reported. A 63-year-old woman underwent right nephrectomy 9 years previously and experienced no recurrence during follow-up. A gradually enlarging ulcerative tumor over the bulb of the duodenum and four subsequent episodes of massive bleeding from this tumor occurred between June 2001 and March 2002. The patient underwent surgery in April 2002 for intractable bleeding from the tumor. Renal cell carcinoma metastasis to the duodenum was confirmed from the surgical specimen. Upper gastrointestinal bleeding due to malignancy is very rare and the duodenum is the least frequently involved site. Furthermore, a solitary late renal cell carcinoma metastasis 9 years after a nephrectomy is extremely uncommon. This case suggests that life-long follow-up of renal cell carcinoma patients is necessary, owing to unpredictable behavior and the possibility of long disease-free intervals. In nephrectomized patients suffering from gastrointestinal bleeding, complete evaluation, especially endoscopic examination, is indicated. The possibility of late recurrent renal cell carcinoma metastasis to the gastrointestinal tract should be kept in mind, although it is rare. If the patient is fit for surgery, metastatectomy is the first choice of treatment. PMID- 15124900 TI - Schwannoma of the nasal septum: a case report. AB - Schwannoma is a neurogenic tumor arising from the sheath of myelinated nerves. Only 4% of schwannomas located in the head and neck region involve the sinonasal tract, and those arising from the nasal septum are exceedingly rare. We report a case of a 55-year-old male who presented with only right nasal obstruction. Computerized tomography and clinical examination suggested a benign tumor before the tumor was completely removed by transnasal endoscopic surgery. When encountering a mass in the nasal cavity, schwannoma should be borne in mind in the differential diagnosis. We discuss the clinical presentation, histologic features, differential diagnosis, and therapeutic options for such a rare lesion. PMID- 15124901 TI - Expression of 60 kDa heat shock protein (Hsp60) on plasma membrane of Daudi cells. AB - In Daudi cells, a fraction of the 60 kDa heat shock protein (Hsp60), which is typically a mitochondrial protein, is located on cell membrane. This was demonstrated by the recovery of biotinylated Hsp60 in the anti-Hsp60 immunoprecipitate obtained from cells in which surface-exposed proteins were selectively labeled with biotin. In further experiments, isolated membrane proteins (obtained by two different biochemical methods) were probed in Western blot with two antibodies (N-20 and K-19) directed against different epitopes located, respectively, at the amino- and at the carboxyl-terminus of the Hsp60. Both these antibodies recognized, among the isolated membrane proteins, a unique band with an electrophoretic mobility identical to that of the cytoplasmic Hsp60, thus demonstrating that Hsp60 is present on cell surface as an intact, full length, protein. FACS analysis, performed with the same two highly specific anti Hsp60 antibodies, confirmed that both the N-terminus and the C-terminus of the Hsp60 are exposed outside the cell and are accessible for recognition by the corresponding antibody. Moreover, quantitative analysis of the data showed that constitutive cell surface expression of the Hsp60 is limited to a small fraction (about 10%) of the whole cell population. PMID- 15124902 TI - Energy metabolism during cutaneous wound healing in immunocompromised and aged rats. AB - Cutaneous cells primarily depend upon carbohydrate metabolism for their energy requirement during healing process. But, it may be greatly hampered during various pathological and altered physiological conditions. The present study was therefore undertaken to investigate the intermediate steps of energy metabolism by measuring enzyme activities in the granulation tissues of immunocompromised and aged rats following excision-type of cutaneous injury. The activities of key regulatory enzymes hexokinase (HK), phosphofructokinase (PFK), lactate dehydrogenase (LDH), citrate synthase (CS) and glucose-6 phosphate dehydrogenase (G6PD) have been monitored in the wound tissues of immunocompromised and aged rats at different time intervals (2, 7, 14 and 21 days) of postwounding. The activities of HK and CS were found significantly decreased both in immunocompromised and aged rats as compared to control subjects. However G6PD exhibited an elevated activity at early stage followed by a decreased activity at later phase of healing both in immunocompromised and aged rats. The PFK and LDH demonstrated an upward trend in immunocompromised rats but a decreasing trend in aged rats. Thus, the results suggest that significant alterations in the activities of energy metabolizing enzymes in the granulation tissues in both immunocompromised as well as in aged rats may overall affect the energy availability for cellular activity needed for repair process. Hence, this may perhaps be one of the factor responsible for impaired healing in these subjects. PMID- 15124903 TI - Comparison of inhibitory effects of oxygen radicals and calf serum protein on surfactant activity. AB - The effects of the reactive oxygen species (ROS) superoxide anion (O2*-) and hydroxyl radical (*OH) on the surface tension lowering properties of bovine lipid extract surfactant (BLES) were compared to the effects of calf serum protein (CSP) in a captive bubble surfactometer (CBS). O2*- was generated from xanthine/xanthine oxidase (X/XO), and *OH was generated by the Fenton reaction. ROS were demonstrated by electron spin resonance (ESR) using 5,5-dimethyl-1 pyrroline-N-oxide (DMPO) as the spin trap. Lipid peroxidation was measured using the thiobarbituric acid method. *OH had broad inhibitory effects on surface tension parameters, including adsorption, minimum surface tension, percentage film area change and film compressibility. O2*- showed inhibitory effects on adsorption, film area change and film compressibility but had no significant effect on minimum surface tension. Both O2*- and *OH treatment were associated with a large 'squeezeout' plateau around 20-25 mN/m in the surface tension-area relation, indicating poor film organization during the compression phase. At the concentrations used, ROS were associated with lipid peroxidation of BLES, which also demonstrated radical scavenging properties. Calf serum protein produced inhibitory effects on adsorption, minimum surface tension and percentage film area change that were quantitatively similar to those produced by *OH. The effects on film compression were significantly greater and qualitatively different from those seen with either O2*- or *OH. We conclude that the inhibition of BLES surface activity by ROS and inhibitory proteins can be distinguished in the captive bubble surfactometer and, particularly, by changes in the film compressibility modulus. PMID- 15124904 TI - Glucuronic acid is a novel inducer of heat shock response. AB - The elevated expression of 70 kDa heat shock protein (Hsp70) induces resistance to stress-induced apoptosis. We have screened a variety of natural products for their ability to enhance Hsp70 expression as anti-apoptotic agent. We found that glucuronic acid (GA) induced the synthesis of Hsp70 and that cells pretreated with GA were significantly tolerant to stress including heat shock and hydrogen peroxide. We also found that GA induces the production of reactive oxygen species (ROS), a process inhibited by NADPH oxidase inhibitor, diphenyleneiodonium chloride (DPI) and antioxidant N-acetylcysteine (NAC). GA-induced ROS production was also inhibited in RacN17 cell line overexpressing a dominant negative mutant of Rac1. Furthermore, GA treatment induces MAPKs activation (SAPK/JNK and p38) and Hsp70 expression in ROS dependent manner, suggesting that GA turns on the signaling pathway by generation of ROS through Rac1. We analyzed the profiles of newly synthesized proteins by GA with 2-dimensional gel electrophoresis and MALDI TOF MS and found that two families of proteins were expressed by GA. One was similar to the protein family synthesized by heat shock (Hsp70, Hsp73, Hsp65, Hsp90, vimentin, tubulin, Ras homolog); and the other was a family of protein specific to GA (calreticulin, annexin III, thioredoxin peroxidase). These results suggest that GA-induced stress responses are mediated by ROS generation and are similar, in part, to heat shock-induced responses and GA can be possibly adopted for the protecting agent from cell death. PMID- 15124905 TI - Inhibition of Ras-GTPase, but not tyrosine kinases or Ca2+/calmodulin-dependent protein kinase II, improves recovery of cardiac function in the globally ischemic heart. AB - The signaling pathways involved in ischemic heart disease are not well characterized. In this study, the roles of Ras-GTPase, tyrosine kinases (TKs) and Ca2+/calmodulin-dependent protein kinase II (CaMKII) in global ischemia and reperfusion (I/R) in a perfused rat heart model were investigated and compared to beneficial effects produced by preconditioning (PC). A 40 min episode of global ischemia followed by a 30 min reperfusion in perfused rat hearts produced significantly impaired cardiac function, measured as left ventricular developed pressure (Pmax) and left ventricular end-diastolic pressure (LVEDP), and impaired coronary hemodynamics, measured as coronary flow (CF) and coronary vascular resistance (CVR). Hearts from male Wistar rats pre-treated with the tyrosine kinase inhibitor, genistein (1 mg/kg/day for 6 days), or the CaMKII inhibitor, KN 93 (578 ng/min for 6 days), produced detrimental effects on recovery of cardiac function and coronary hemodynamics. In contrast, pre-treatment with Ras-GTPase inhibitor FPT III (232 ng/min for 6 days) significantly enhanced cardiac recovery in terms of left ventricular contractility and coronary vascular hemodynamics. Treatment with FPT III also significantly reduced expression of the sodium hydrogen exchanger-1 (NHE-1) which was elevated during I/R as detected by Western blotting. These data suggest that TKs and CaMKII are involved in signaling pathways leading to recovery from cardiac ischemia, whereas activation of Ras GTPase signaling pathways are critical in the development of cardiac dysfunction due to I/R. PMID- 15124906 TI - Caspase-3: its potential involvement in Cr(III)-induced apoptosis of lymphocytes. AB - In this study, we have examined the role of caspase-3 in apoptosis of lymphocytes induced by the chromium(III) complexes viz. tris-(1,10 phenanthroline)chromium(III) chloride (Cr(III)-phen) and trans-diaqua[1,3 bis(salicylideneamino)propanechromium(III)] perchlorate (Cr(III)-salprn). Evidence for caspase-3 activation and poly(ADP-ribose) polymerase (PARP) cleavage in lymphocytes exposed to Cr(III) complexes is revealed through Western blotting analysis. Blocking the activity of caspase-3 with z-DEVD-fmk, prevents apoptosis as evidenced through [3H]-thymidine incorporation, DNA fragmentation assay and measurement of sub-G1 cells by flow cytometry. Pretreatment of lymphocytes with free radical scavengers completely attenuates the activity of caspase-3 suggesting that reactive oxygen species (ROS) are upstream activators of caspase 3. Preincubation of lymphocytes with PP2, a selective Src-family tyrosine kinase inhibitor, abolishes the activation of caspase-3 indicating that Src-family tyrosine kinases viz. p56lck, p59fyn and p53/56lyn are mediators of caspase-3 activation during Cr(III) exposure. Collectively, our findings support a plausible mechanism in which Cr(III) mediates ROS generation that precedes the up regulation of p56lck, p59fyn and p53/56lyn which eventually activates caspase-3 to promote apoptotic cell death of lymphocytes. To our knowledge, this is the first report suggesting the importance of Src-family tyrosine kinases for the activation of caspase-3 in metal-induced apoptotic cell death. PMID- 15124907 TI - Impaired Na+,K+-ATPase activity as a mechanism of reactive nitrogen species induced cytotoxicity in guinea pig liver exposed to lipopolysaccharides. AB - In animal models of endotoxin, the excess production of NO and the reactive nitrogen species (RNS), are potent oxidant and nitrating agents, lead to lipid peroxidation, apoptosis, tissue dysfunction and injury and inactivate enzymes in many cell types. Although liver functions are well known to deteriorate following bacterial infection, the underlying specific mechanism(s) remain a matter of considerable debate. Therefore, the aim of the present study was to determine the in vivo effect of bacterial lipopolysaccharides (LPS) on Na+,K+-ATPase activity of guinea pig liver, and to investigate the possible contribution of RNS by measuring of iNOS activity and 3-nitrotyrosine (nTyr) levels. Liver Na+,K+-ATPase activity were maximally inhibited 6 h after LPS injection (p < 0.001 ). nTyr was not detectable in liver of normal control animals, but was detected markedly in LPS exposed animals. LPS treatment significantly increased iNOS activity of liver (p < 0.001). The regression analysis revealed a very close correlation between Na+,K+-ATPase activity and nTyr levels of LPS treated animals (r = -0.863, p < 0.001). Na+, K+-ATPase activity were also negatively correlated with iNOS activity (r = -0.823, p < 0.003) in inflamed tissues. Our results have strongly suggested that bacterial LPS disturbs activity of membrane Na+,K+-ATPase that may be an important component leading to the pathological consequences such as hepatocyte cell loss and dysfunction in which the production of RNS are increased as in the case of LPS challenge. PMID- 15124908 TI - Apolipoprotein C3 SstI polymorphism in the risk assessment of CAD. AB - Various population studies have reported the association of rare S2 allele of apolipoprotein C3 (APOC3) SstI polymorphism with hypertriglyceridemia (HTG) and coronary artery disease (CAD). We were the first to report an association of S2 allele with high triglyceride (TG) levels in healthy volunteers from Northern India. Since HTG is suggested to be a predominant risk factor for CAD among Indians, we have elucidated the relationship of APOC3 SstI polymorphism with the lipid profile and CAD. A total of 158 patients with > or = 70% stenosis in one or more coronary artery (angiographically proven CAD patients), 35 subjects with < 70% stenosis (NCAD) and 151 normal controls (free of heart disease) from Northern plains of India were recruited in the study. DNA samples were analyzed by polymerase chain reaction (PCR) followed by SstI digestion. Lipid profile was estimated by enzymatic kit. We found a strong association of S2 allele with high TG levels, which was more significant in patients. Prevalence of S2 allele in normal controls and CAD patients were comparable, despite the fact that mean TG level was significantly higher in patients. A greater insight into this observation revealed that the prevalence of high TG, if not coupled with other risk factors (like high total cholesterol, low HDL), was comparable in patients and controls. Thus, our study reveals that rare S2 allele may be employed as a susceptibility marker for high TG. However, high TG or S2 allele alone may not contribute to the etiology of CAD. PMID- 15124909 TI - Evidence of apoptosis in human diabetic kidney. AB - Diabetic nephropathy is characterized by an early period of renal growth with glomerular and tubular cell hypertrophy, but this is followed by progressive glomerulosclerosis and tubulointerstitial fibrosis, associated with loss of renal tissue. We studied whether apoptotic cell death occurs in human diabetic nephropathy. Percutaneous renal biopsy samples were obtained from five patients with diabetic nephropathy who were receiving insulin and/or angiotensin converting enzyme inhibitor therapy. Apoptosis was determined by the presence of DNA fragmentation, detected by in situ TUNEL staining, and by characteristic features on electron microscopy, such as chromatin condensation. Apoptosis was present in all five biopsy specimens, either in epithelial cells of the proximal or distal tubules, or in endothelial cells or interstitial cells. No apoptosis was detected in cells of the glomeruli. The present study provides evidence for apoptosis in human diabetic kidney, and suggests a role for apoptosis in the gradual loss of renal mass. PMID- 15124910 TI - TBP-associated factor 1 overexpression induces tolerance to Doxorubicin in confluent H9c2 cells by an increase in cdk2 activity and cyclin E expression. AB - Doxorubicin (DOX) is a DNA topoisomerase II inhibitor widely used in anticancer treatment, however, it can lead to irreversible cardiac damage with severe debilitation. TBP-binding associated factor 1 (TAF1) is increased in DOX damaged hearts in vivo and in cardiomyocytes in vitro. To identify the functional role for TAF1 in DOX-treated heart we overexpressed wild type and mutant TAF1 in H9c2 cells. Overexpression of wild-type TAF1, but not N-terminal kinase domain mutants, increased tolerance to DOX in confluent cells. DOX treatment can cause prolonged G1 arrest. We found increased cdk2 activity coupled to increased cyclin E protein and decreased p21(waf1Cip1) and p27(Kip1) protein to correlate only with increased DOX tolerance and wild-type TAF1. DOX sensitivity was restored when the cdk2-inhibitor Roscovitine was co-administered with DOX. Overexpression of cdk2-alone increased resistance to DOX. Thus, TAF1 induced DOX tolerance in confluent cells through an increase in cdk2 activity is directed by the TAF1 N terminal domain. These studies suggest new avenues for myocardial protection against DOX toxicity and suggest a role for cdk2 in chemorefractory cells. PMID- 15124911 TI - Cell survival signalling in heart derived myofibroblasts induced by preconditioning and bradykinin: the role of p38 MAP kinase. AB - Fibroblasts possess receptors for compounds released during ischemia, including bradykinin. The aims of the present study were to investigate tyrosine kinase and p38 MAP kinase signalling in heart derived myofibroblasts in response to bradykinin and preconditioning ischemia. Fibroblasts from neonatal rat hearts were subjected to pharmacological agents and/or simulated ischemia. Cell viability was measured by the conversion of a tetrazolium salt to its formazan derivative. Preconditioning with 30 min of simulated ischemia followed by 30 min recovery resulted in an 85.4% +/- 7.8% increase in cell survival above that of cells treated with prolonged ischemia alone. Cells treated with bradykinin showed a 35% +/- 7.9 increase in cell survival after lethal ischemia. The B2 receptor antagonist Hoe 140 blocked the protective effect of bradykinin, but did not block preconditioning. The K(ATP) channel blocker glibenclamide and the mitochondria specific K(ATP) blocker 5, hydroxydecanoate, abolished the cytoprotection induced by both preconditioning and bradykinin. The non specific tyrosine kinase inhibitor genistein also abolished the cytoprotection. Effective blockade of cytoprotection was obtained with K(ATP) channel blockers and the tyrosine kinase inhibitor when these compounds were given prior to the preconditioning stimulus and not during the lethal insult. The stress activated protein kinase p38 MAP kinase was investigated by Western blotting and by the use of a specific inhibitor (SB203580). Preconditioning reduced phospho-p38 MAP kinase; in contrast, bradykinin administration markedly increased phosphorylation of p38 MAP kinase. SB203580 protected cells from lethal simulated ischemia. In conclusion, cell survival-signalling pathways activated by bradykinin or simulated ischemia in heart fibroblasts protect via the opening of K(ATP) channels and are independent of the stress-activated p38 MAP kinase and/or related to inhibition of this kinase. PMID- 15124912 TI - Effects of chronic quercetin treatment on antioxidant defence system and oxidative status of deoxycorticosterone acetate-salt-hypertensive rats. AB - We investigated the potential of chronic administration of an oral daily dose (10 mg/kg) of the dietary flavonoid quercetin to prevent hypertension and oxidative stress induced by deoxycorticosterone acetate (DOCA)-salt in rats. We have compared its effects to those produced by the well-known anti-hypertensive drug verapamil, administered orally (20 mg/kg/day). Quercetin and verapamil treatments reduced systolic blood pressure of DOCA-salt rats in approximately 67.6 and 63.3% respectively, producing no effect in control animals. Both drugs reduced significantly hepatic and renal hypertrophy induced by DOCA-salt administration, while only quercetin prevented cardiac hypertrophy. Decreased endothelium dependent relaxation to acetylcholine of aortic rings from DOCA-salt-treated rats was improved by quercetin, but verapamil only enhanced it in the presence of superoxide dismutase (SOD) plus catalase. Increased plasma and heart thiobarbituric acid reactive substances (TBARS) and total glutathione (GSH) levels in liver and heart, decreased liver glutathione peroxidase (GPX) and liver and kidney glutathione transferase (GST) activities were observed in DOCA-salt treated rats compared to the control animals. The antihypertensive effect of quercetin was accompanied by normalisation of plasma TBARS values, improvement of the antioxidant defences system in heart and liver, restoring total GSH levels in both organs and altered liver GST and GPX activities, and improving kidney GST activity. Verapamil treatment only restored GSH levels in heart, having no effect on other alterations induced by DOCA-salt chronic administration in the antioxidant defences analysed. In conclusion, quercetin shows both antihypertensive and antioxidant properties in this model of mineralocorticoid hypertension, while verapamil exhibits only antihypertensive effects. PMID- 15124913 TI - Metabolism of tritiated D-glucose anomers in rat erythrocytes. AB - It was recently proposed that alpha-D-glucose 6-phosphate may undergo enzyme-to enzyme channelling between glucokinase and phosphoglucoisomerase in rat pancreatic islets. The present study aims at exploring whether a different situation prevails in cells deprived of glucokinase, namely in erythrocytes. At anomeric equilibrium, the ratio between D-[2-3H]glucose and D-[5-3H]glucose conversion to 3HOH was lower in rat erythrocytes incubated for 60 min at 4 degrees C in the presence of 2.8 mM, rather than 8.3 mM, D-glucose. This coincided with both a greater relative increase in beta-D-[5-3H]glucose, as compared to alpha-D-[5-3H]glucose, conversion to 3HOH and an increase in the beta/alpha ratio for 3HOH generation from D-[5-3H]glucose in response to an increase in the anomeric concentration from 2.8 to 8.3 mM, the suppression of the difference between the beta/alpha ratios for 3HOH generation from D-[2-3H]glucose and D-[5-3H]glucose in the erythrocytes incubated at 8.3 mM, as distinct from 2.8 mM, alpha- and beta-D-glucose, and a [2-3H]/[5-3H] ratio for 3HOH generation lower than unity in erythrocytes exposed to alpha-D-glucose but not significantly different from unity in the presence of beta-D-glucose. These findings emphasize the relevance of alpha-D-glucose 6-phosphate channelling between hexokinase and phosphoglucoisomerase as a determinant of the difference between D-[2-3H]glucose and D-[5-3H]glucose conversion to 3HOH, and reveal that the regulation of such a tunnelling process by the concentration of the D-glucose represents, in rat erythrocytes, a mirror image of that observed in rat pancreatic islets. The regulation of this process thus tightly depends on the identity of the hexokinase enzyme mainly responsible for the phosphorylation of D-glucose in distinct cell types. PMID- 15124914 TI - Permeability of fructose-1,6-bisphosphate in liposomes and cardiac myocytes. AB - Fructose-1,6-bisphosphate (FBP) helps preserve heart and other organs under ischemic conditions. Previous studies indicated that it can be taken up by various cell types. Here we extended observations from our group that FBP could penetrate artificial lipid bilayers and be taken up by cardiac myocytes, comparing the uptake of FBP to that of L-glucose. Using liposomes prepared by the freeze-thaw method, FBP entered about 200-fold slower than L-glucose. For liposomes of either soybean or egg lipids, 50 mM FBP enhanced the permeability of FBP itself, with little effect on general permeability (measured by uptake of L glucose). In experiments with isolated cardiac myocytes at 21 degrees C, FBP uptake exceeded the uptake of L-glucose by several fold and appeared to equilibrate by 60 min. There was both a saturable component at micromolar levels and a nonsaturable component which dominated at millimolar levels. The saturable component was inhibited by Pi and by other phosphorylated sugars, though with lower affinity than FBP. Both saturable and nonsaturable uptakes were also observed at 3 degrees C. The results indicate that FBP enters myocytes not by simple penetration through the lipid bilayer, but via at least two distinct protein-dependent processes. The uptake could lead to intracellular effects important in hypothermic heart preservation. PMID- 15124915 TI - Liver fatty acid binding protein expression enhances branched-chain fatty acid metabolism. AB - Although liver fatty acid binding protein (L-FABP) is known to enhance uptake and esterification of straight-chain fatty acids such as palmitic acid and oleic acid, its effects on oxidation and further metabolism of branched-chain fatty acids such as phytanic acid are not completely understood. The present data demonstrate for the first time that expression of L-FABP enhanced initial rate and average maximal oxidation of [2,3-3H] phytanic acid 3.5- and 1.5-fold, respectively. This enhancement was not due to increased [2,3-3H] phytanic acid uptake, which was only slightly stimulated (20%) in L-FABP expressing cells after 30 min. Similarly, L-FABP also enhanced the average maximal oxidation of [9,10 3H] palmitic acid 2.2-fold after incubation for 30 min. However, the stimulation of L-FABP on palmitic acid oxidation nearly paralleled its 3.3-fold enhancement of uptake. To determine effects of metabolism on fatty acid uptake, a non metabolizable fluorescent saturated fatty acid, BODIPY-C16, was examined by laser scanning confocal microscopy (LSCM). L-FABP expression enhanced uptake of BODIPY C16 1.7-fold demonstrating that L-FABP enhanced saturated fatty acid uptake independent of metabolism. Finally, L-FABP expression did not significantly alter [2,3-3H] phytanic acid esterification, but increased [9,10-3H] palmitic acid esterification 4.5-fold, primarily into phospholipids (3.7-fold) and neutral lipids (9-fold). In summary, L-FABP expression enhanced branched-chain phytanic acid oxidation much more than either its uptake or esterification. These data demonstrate a potential role for L-FABP in the peroxisomal oxidation of branched chain fatty acids in intact cells. PMID- 15124916 TI - Estradiol-mediated internalisation of the non-activated estrogen receptor from the goat uterine plasma membrane: identification of the proteins involved. AB - An indirect approach has been made to study the molecular details associated with the estradiol-induced internalisation of the non-activated estrogen receptor (naER) from the goat uterine plasma membrane. The internalisation of naER appears to be an energy dependent process. Exposure of the plasma membrane to estradiol results in the activation of a Mg2+ dependent ATPase associated with the membrane fraction. Presence of quercetin in the medium prevented the activation of the Mg2+ ATPase as well as the dissociation of naER from the plasma membrane. Using isolated plasma membrane preparations it has been possible to identify the proteins which interact with naER during various stages of its internalisation. The main proteins identified are: (1) a 58 kDa protein, p58, which apparently recognizes the nuclear localization signals on the naER and transports it to the nucleus: (2) hsp70: (3) hsp90, the functional roles of which remain unknown at this stage; (4) a 50 kDa protein associated with the clathrin coated vesicles, presumed to be involved in recognizing the tyrosine based internalisation signals on the naER; (5) actin which mediates the plasma membrane-to-nucleus movement of the naER-p58 complex. PMID- 15124917 TI - Proteins which mediate the nuclear entry of goat uterine non activated estrogen receptor (naER) following naER internalization from the plasma membrane. AB - The nuclear transport of the internalised naER is influenced by a 58 kDa protein, p58, that appears to recognize the nuclear localization signals on the naER. At the nuclear pore complex the naER-p58 complex binds to a 62 kDa protein, p62; p58 recognizes p62 in this interaction. It is further observed that p62 gets 'docked' at a 66 kDa nuclear pore complex protein, npcp66. The nuclear entry of naER is an ATP-dependent process. An ATP-dependent biphasic nuclear entry of naER, has been observed. It is possible that the docking of p58-naER complex at the nuclear pore complex and the eventual nuclear entry of naER following its dissociation from the p58 are influenced by two different ranges in the concentration of ATP. In this process, it appears that, the nuclear entry requires an additional quantum of energy, provided by the hydrolysed ATP, in contrast to the energy requirement associated with, the nuclear 'docking' event. PMID- 15124918 TI - Pure pressure stress increased monocarboxylate transporter in human aortic smooth muscle cell membrane. AB - Lactate is formed and utilized continuously under fully aerobic conditions. Lactate is oxidized actively at all times, especially during exercise. Family of monocarboxylate transport proteins (MCTs) that are differentially expressed in cells and tissues accomplishes the facilitated transport of lactate across membranes. Previously we reported that there is MCT1 in blood circulation. We also reported the pressure stress stimulated cell proliferation in aortic smooth muscle cells (HASMC). In this experiment we attempted to prove the existence of MCT1 in HASMC and to clarify the effect of pressure stress on MCT1 localization in HASMC. We determined succinate dehydrogenase (SDH) activity as a marker of energy metabolism in cells. SDH activity was increased by pressure stress. Lactate enhanced the SDH activity under pressure stress (160 mmHg for 3 h) as dose dependent manner. On the other hand, lactate excretion was suppressed by the addition of lactate. We could detect MCT1 in the cytosolic and the membrane fractions of HASMC. The pressure stress increased MCT1 in the membrane fraction in the presence of extracellular lactate. In summary, we proved the existence of MCT1 in HASMC. Pressure stress changed the localization of MCT1. The increased membranous MCT1 may transport lactate for energy metabolism in cells. PMID- 15124919 TI - Development and application of Na+/Ca2+ exchange inhibitors. AB - The Na+/Ca2+ exchanger (NCX) is an ion transporter that exchanges Na+ and Ca2+ in either Ca2+ efflux or Ca2+ influx mode, depending on the ion gradients across the plasma membrane and the membrane potential. In heart, smooth muscle cells, neurons, and nephron cells, the NCX is thought to play an important role in the regulation of intracellular Ca2+ concentration. Recently, a novel selective inhibitor (KB-R7943 and SEA0400) of the Ca2+ influx mode of the NCX has been developed. NCX inhibitor is expected to be a pharmaceutical agent that offers effective protection against ischemia/reperfusion injury in several organs such as heart and kidney. Here, we summarize pharmacological profiles of KB-R7943 and SEA0400, the molecular mechanism of its action, and its future prospect as a novel pharmaceutical agent. PMID- 15124920 TI - Intracellular signaling pathways for norepinephrine- and endothelin-1-mediated regulation of myocardial cell apoptosis. AB - Accumulating data support the idea that apoptosis in cardiac myocytes, in part, contributes to the development of heart failure. Since a number of neurohormonal factors are activated in this state, these factors may be involved in the positive and negative regulation of apoptosis in cardiac myocytes. Norepinephrine is one such factor and induces apoptosis in cardiac myocytes via a beta adrenergic receptor pathway. beta-adrenergic agonist-induced apoptosis in cardiac myocytes is dependent on the activation of the cAMP/protein kinase A pathway. Interestingly, the activation of this pathway protects PC12 cells from apoptosis, suggesting that cAMP/protein kinase A regulates apoptosis in a cell type-specific manner. Another neurohormonal factor activated in heart failure is endothelin-1, which acts as a potent survival factor against myocardial cell apoptosis. Intracellular signaling pathways for endothelin-1-mediated protection include activation of MEK-1 /ERK1/2 and PI3 kinase. In addition to these protective pathways common among cell types, endothelin- activates the calcium-activated phosphatase calcineurin, which is necessary for the nuclear import of NFAT transcription factors. These factors interact with the cardiac-restricted zinc finger protein GATA-4 and induce transcription and expression of anti-apoptotic molecule bcl-2. Thus, myocardial cell apoptosis is regulated by pathways unique to cardiac myocytes as well as by those common among cell types. It should be further determined whether agents that specifically block myocardial cell apoptosis will attenuate the progression of heart failure. PMID- 15124921 TI - Akt and Ca2+ signaling in endothelial cells. AB - The protein kinase Akt participates in such important functions of endothelial cells as nitric oxide production and angiogenesis, activities that involve changes in cytosolic Ca2+ concentration. However, it is not known if activation of Akt is itself involved in the regulation of Ca2+ signals produced in these cells. The objective of this study was to examine if Akt is involved in the regulation of Ca2+ signaling in endothelial cells. Agonist-stimulated Ca2+ signals, assessed using fura-2, were compared in porcine aortic endothelial cells under control conditions or conditions in which Akt was blocked either by different inhibitors of phosphatidylinositol 3-kinase (PI3 kinase)/Akt or by transient expression of a dominant-negative form of Akt (dnAkt). We found that the release of intracellular Ca2+ stores stimulated by bradykinin or thapsigargin is not affected by the PI3 kinase inhibitors LY294002 and wortmannin, or by expression of dnAkt. LY294002 dose-dependently inhibits store-operated Ca2+ entry, an effect not seen with wortmannin. Expression of dnAkt has no effect on store-operated Ca2+ entry. We conclude that Akt is not involved in the regulation of agonist-stimulated Ca2+ signals in endothelial cells. The compound LY294002 inhibits store-operated Ca2+ entry in these cells by a mechanism independent of PI3 kinase/Akt inhibition. PMID- 15124922 TI - Cardiomyocytes undergo cells division following myocardial infarction is a spatially and temporally restricted event in rats. AB - Dividing cardiomyocytes are observed in autopsied human hearts following recent myocardial infarction, however there is a lack of information in the literature on the division of these cells. In this study we used a rat model to investigate how and when adult mammalian cardiomyocytes proliferate by cell division after myocardial infarction. Myocardial infarction was induced in Wistar rats by ligation of the left coronary artery. The rats were sacrificed periodically up to 28 days following induced myocardial infarction, and the hearts subjected to microscopic investigation. Cardiomyocytes entering the cell cycle were assayed by observation of nuclear morphology and measuring expression of Ki-67, a proliferating cell marker. Ki-67 positive cardiomyocytes and dividing nuclei were observed initially after 1 day. After 2 days dividing cells gradually increased in number at the ischemic border zone, reaching a peak increase of 1.12% after 3 days, then gradually decreasing in number. Dividing nuclei increased at the ischemic border zone after 3 days, peaked by 0.14% at day 5, and then decreased. In contrast, Ki-67 positive cells and dividing nuclei were limited in number in the non-ischemic area throughout all experiments. In conclusion, mitogenic cardiomyocytes are present in the adult rat heart following myocardial infarction, but were spatially and temporally restricted. PMID- 15124923 TI - Electrical and ionic abnormalities in the heart of cardiomyopathic hamsters: in quest of a new paradigm for cardiac failure and lethal arrhythmia. AB - Cardiomyopathy is primary degenerative disease of myocardium, which leads to cardiac failure and lethal arrhythmia. An appropriate model animal of a particular disease is, in general, greatly helpful for better understanding of its pathogenesis. In 1962, a naturally occurring mutant line of Syrian hamster named BIO1.50 was reported, which inherited cardiomyopathy and muscular dystrophy as autosomal recessive mode with 100% penetrance. To date, several sublines of cardiomyopathic hamsters (CM hamsters) have been derived. The genomic deletion of delta-sarcoglycan, a member of dystrophin-associated proteins, was demonstrated to be the common genetic cause of CM hamsters in 1997. Over the past 40 years, hundreds of papers have been published on the pathophysiological aspects of CM hamsters. The aim of this paper is to annotate every one of the CM hamsters with its historical background and then summarize the previous findings on CM hamsters with special focus on electrical and ionic properties. This review article is expected to serve as a basis to build up a new paradigm for the pathogenesis of cardiac failure and severe arrhythmia. PMID- 15124924 TI - Regulatory molecules for coronary expressions of VEGF and its angiogenic receptor KDR in hypoestrogenic middle-aged female rats. AB - We studied the effects of estrogen deprivation and replacement on the protein and gene expression levels molecules that can be considered to be essential for coronary angiogenesis in middle-aged female rats. The animals were subjected to sham operation, ovariectomy, or ovariectomy with estrogen replacement therapy (ERT). Following ovariectomy, protein and gene expressions of vascular endothelial growth factor (VEGF) and its angiogenic receptor (KDR) showed a marked decline in coronary vessels, as determined by immunohistochemistry and in situ hybridization. ERT resulted in restoration of the ovariectomy-induced changes to intact levels. The coronary expression level of basic fibroblast growth factor was unaffected by estrogen deprivation or treatment. The changes in VEGF and KDR expressions were strongly associated with those in endothelial nitric oxide synthase (eNOS) expression in coronary vessels. Moreover, the age- and gender-dependent accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha) protein appeared to be a determinant molecule of VEGF expression in middle-aged female rats. We reached a conclusion that the VEGF-KDR system plays a key role in coronary angiogenesis in hypoestrogenic elderly women and is critically regulated by estrogen, eNOS and HIF-1alpha. PMID- 15124926 TI - Induction of heat shock protein 72 in the failing heart is attenuated after an exposure to heat shock. AB - Induction of heat shock protein (Hsp) 72 in the right ventricular muscle of the rat with heart failure following acute myocardial infarction (AMI) was examined. AMI was induced by the left coronary artery ligation (CAL). The animals at the 8th, but not 2nd, week after CAL revealed a decrease in cardiac output index (COI), suggesting that heart failure had developed by 8 weeks after CAL. Increases in the right ventricular developed pressure and the ratios of right ventricle/body weight and lung/body weight at the 2nd and 8th weeks showed the development of the right ventricular hypertrophy. After measurement of hemodynamic parameters, the hearts isolated from animals at the 2nd and 8th weeks after CAL (2w- and 8w-CAL hearts, respectively) were perfused and subjected to heat shock (at 42 degrees C, for 15 min) followed by 6-h perfusion. At the end of perfusion, Hsp72 content in the left ventricle without infarct area (viable LV) and the right ventricle (RV) was determined by the Western immunoblotting method. The production of myocardial Hsp72 in the viable LV and RV of the 2w-CAL heart increased after an exposure to heat shock. In contrast, induction of Hsp72 in the viable LV and RV of the 8w-CAL heart was blunted. The results suggest that the development of heart failure following AMI may result in a decrease in the ability for Hsp72 induction not only in the viable LV but also in the RV, leading to contractile dysfunction of the heart. PMID- 15124925 TI - Myosin phosphatase: structure, regulation and function. AB - Phosphorylation of myosin II plays an important role in many cell functions, including smooth muscle contraction. The level of myosin II phosphorylation is determined by activities of myosin light chain kinase and myosin phosphatase (MP). MP is composed of 3 subunits: a catalytic subunit of type 1 phosphatase, PPlc; a targeting subunit, termed myosin phosphatase target subunit, MYPT; and a smaller subunit, M20, of unknown function. Most of the properties of MP are due to MYPT and include binding of PP1c and substrate. Other interactions are discussed. A recent discovery is the existence of an MYPT family and members include, MYPT1, MYPT2, MBS85, MYPT3 and TIMAP. Characteristics of each are outlined. An important discovery was that the activity of MP could be regulated and both activation and inhibition were reported. Activation occurs in response to elevated cyclic nucleotide levels and various mechanisms are presented. Inhibition of MP is a major component of Ca2+-sensitization in smooth muscle and various molecular mechanisms are discussed. Two mechanisms are cited frequently: (1) Phosphorylation of an inhibitory site on MYPT1, Thr696 (human isoform) and resulting inhibition of PP1c activity. Several kinases can phosphorylate Thr696, including Rho-kinase that serves an important role in smooth muscle function; and (2) Inhibition of MP by the protein kinase C-potentiated inhibitor protein of 17 kDa (CPI-17). Examples where these mechanisms are implicated in smooth muscle function are presented. The critical role of RhoA/Rho-kinase signaling in various systems is discussed, in particular those vascular smooth muscle disorders involving hypercontractility. PMID- 15124927 TI - Beneficial effects of olmesartan, a novel angiotensin II receptor type 1 antagonist, upon acute autoimmune myocarditis. AB - Excess amount of cytokine produced by inflammatory stimuli contributes to the progression of myocardial damage in myocarditis. Some angiotensin II receptor type 1 antagonists are reported to inhibit proinflammatory cytokine production in vitro and in vivo. We tested the hypothesis that olmesartan, a novel angiotensin II receptor type 1 antagonist, ameliorated experimental autoimmune myocarditis (EAM) in rats attributing to the suppression of inflammatory cytokines in the heart. We orally administered olmesartan 1, 3, and 10 mg/kg/day to rats with EAM for 3 weeks. The results showed that olmesartan decreased blood pressure significantly compared with the untreated group, but markedly reduced the severity of myocarditis by comparing the heart weight/body weight ratio, pericardial effusion scores, macroscopic scores and microscopic scores. Myocardial interleukin (IL)- 1beta expression by western blotting and IL-1beta positive staining cells by immunohistochemistry were significantly lower in rats with EAM given olmesartan treatment compared with those of rats given vehicle. We conclude that Olmesartan ameliorates acute EAM in rats. The cardioprotection of olmesartan may be due to suppression of inflammatory cytokines dependent of the hemodynamic modifications. PMID- 15124928 TI - Cardioprotective effects of carvedilol on acute autoimmune myocarditis. AB - We investigated whether carvedilol protects against experimental autoimmune myocarditis (EAM) attributing to antioxidant properties. Acute EAM was induced by porcine cardiac myosin in Lewis rats. We orally administered a vehicle, various dosages of carvedilol, metoprolol, or propranolol to rats with EAM for 3 weeks. Three beta-blockers decreased heart rates to the same extent. Carvedilol, but not metoprolol or propranolol, markedly reduced the severity of myocarditis at the two different dosages. Only carvedilol decreased the myocardial protein carbonyl contents, and also decreased the myocardial thiobarbituric acid reactive substance products in rats with EAM. Accordingly, carvedilol protects against acute EAM in rats, and this superior cardioprotective effect of carvedilol to metoprolol and propranolol may be due to the antioxidant properties in addition to the hemodynamic modifications. PMID- 15124929 TI - A structure-activity relationship study of catechol-O-methyltransferase inhibitors combining molecular docking and 3D QSAR methods. AB - A panel of 92 catechol-O-methyltransferase (COMT) inhibitors was used to examine the molecular interactions affecting their biological activity. COMT inhibitors are used as therapeutic agents in the treatment of Parkinson's disease, but there are limitations in the currently marketed compounds due to adverse side effects. This study combined molecular docking methods with three-dimensional structure activity relationships (3D QSAR) to analyse possible interactions between COMT and its inhibitors, and to incite the design of new inhibitors. Comparative molecular field analysis (CoMFA) and GRID/GOLPE models were made by using bioactive conformations from docking experiments, which yielded q2 values of 0.594 and 0.636, respectively. The docking results, the COMT X-ray structure, and the 3D QSAR models are in agreement with each other. The models suggest that an interaction between the inhibitor's catechol oxygens and the Mg2+ ion in the COMT active site is important. Both hydrogen bonding with Lys144, Asn170 and Glu199, and hydrophobic contacts with Trp38, Pro174 and Leu198 influence inhibitor binding. Docking suggests that a large R1 substituent of the catechol ring can form hydrophobic contacts with side chains of Val173, Leu198, Met201 and Val203 on the COMT surface. Our models propose that increasing steric volume of e.g. the diethylamine tail of entacapone is favourable for COMT inhibitory activity. PMID- 15124930 TI - In silico models for the prediction of dose-dependent human hepatotoxicity. AB - The liver is extremely vulnerable to the effects of xenobiotics due to its critical role in metabolism. Drug-induced hepatotoxicity may involve any number of different liver injuries, some of which lead to organ failure and, ultimately, patient death. Understandably, liver toxicity is one of the most important dose limiting considerations in the drug development cycle, yet there remains a serious shortage of methods to predict hepatotoxicity from chemical structure. We discuss our latest findings in this area and present a new, fully general in silico model which is able to predict the occurrence of dose-dependent human hepatotoxicity with greater than 80% accuracy. Utilizing an ensemble recursive partitioning approach, the model classifies compounds as toxic or non-toxic and provides a confidence level to indicate which predictions are most likely to be correct. Only 2D structural information is required and predictions can be made quite rapidly, so this approach is entirely appropriate for data mining applications and for profiling large synthetic and/or virtual libraries. PMID- 15124932 TI - Molecular dynamics simulation of matrix metalloproteinase 2: fluctuations and time evolution of recognition pockets. AB - We report a molecular dynamics simulation study of a zinc-protease--gelatinase A or MMP2--which is a major target for drug design, being involved in tumor metastasis and other degenerative diseases. Two structures have been employed as starting conditions, one based on the crystal of multi-domain proMMP2, the other consisting of the catalytic domain only. The overall fold of the two models is maintained over the 1260 ps trajectory, enabling us to analyze correlations of fluctuations among domains, and to observe the presence of correlations within the catalytic domain in the multi-domain enzyme only, hence due to the presence of hemopexin and fibronectin domains. In the multi-domain protein, two cavities are conserved over the trajectory, one of them pointing to a key region, a crevice surrounding the catalytic zinc. The other one is localized across the three domains of the MMP2 metalloproteinase. These areas are partially covered by the propeptide in the crystal structure of proMMP2. We propose a model of MMP2 collagen interaction that involves both identified cavities and takes into account the inter/intra domain cross-correlations. PMID- 15124931 TI - A molecular modeling study of inhibitors of nuclear factor kappa-B (p50)--DNA binding. AB - Nuclear Factor-kappa B (NF-kappaB) is an inducible transcription factor of the Rel family, and is sequestered in the cytoplasm by the IkappaB family of proteins. NF-kappaB can exist in several dimeric forms, but the p50/p65 heterodimer is the predominant one. Activation of NF-kappaB by a range of stimuli including viral products, and oxidative stress, leads to phosphorylation and proteasome dependent degradation of IkappaB, leading to the release of free NF kappaB. This free NF-kappaB then binds to its target sites (KB sites in the DNA) to initiate transcription. These kappaB sites are also present in the Long Terminal Repeat (LTR) of HIV-1, and hence NF-kappaB (p50 subunit) binding to LTR DNA is critical in viral replication. Targeting direct p50-DNA binding, in this regard, is a novel approach to design anti-HIV gene expression inhibitors, which do not have the problem of resistance unlike in other anti-HIV strategies. The present study is a part of our search for leads for the specific inhibition of p50-DNA binding. We have been experimentally studying different types of these inhibitors, and in this work, we attempted to get a common definition of their structural mechanism onto p50-DNA binding. Using three different classes of inhibitors, we modelled their association with the DNA-Binding Region (DBR) of the p50 subunit of NF-kappaB. Docking studies were carried out using a genetic algorithm based program (GOLD). Further, to compare electrostatic complementarity in the association of the inhibitors with the DBR, Molecular Electrostatic Potentials (MEPs) were generated for the DBR and each inhibitor. The results of docking revealed a strong network of hydrogen bonding interactions for every active inhibitor, and the contrary for the less active ones. Further, the MEPs revealed that the DBR of p50 represents a surface of electropositive potential, and the active inhibitors represent a complementary electronegative surface. With the present modelling study we conclude that the principal properties to be possessed by the new leads against p50-DNA binding should be that of having the ability to make a strong network of hydrogen bonds with the DBR of p50, and preferably, having electronegative potentials in their peripheral surface. PMID- 15124933 TI - Prediction of in vitro metabolic stability of calcitriol analogs by QSAR. AB - The metabolic stability of a drug is an important property for potential drug candidates. Measuring this property, however, can be costly and time-consuming. The use of quantitative structure-activity relationships (QSAR) to estimate the in vitro stability is an attractive alternative to experimental measurements. A data set of 130 calcitriol analogs with known values of in vitro metabolic stability was used to develop QSAR models. The analogs were encoded with molecular structure descriptors computed mainly with the commercial software QikProp and DiverseSolutions. Variable selection was carried out by five different variable selection techniques and Partial Least Squares Regression (PLS) models were generated from the 130 analogs. The models were used for prediction of the metabolic stability of 244 virtual calcitriol analogs. Twenty of the 244 analogs were selected and the in vitro metabolic stability was determined experimentally. The PLS models were able to predict the correct metabolic stability for 17 of the 20 selected analogs, corresponding to a prediction performance of 85%. The results clearly demonstrate the utility of QSAR models in predicting the in vitro metabolic stability of calcitriol analogs. PMID- 15124934 TI - Protein-ligand binding free energy estimation using molecular mechanics and continuum electrostatics. Application to HIV-1 protease inhibitors. AB - A method is proposed for the estimation of absolute binding free energy of interaction between proteins and ligands. Conformational sampling of the protein ligand complex is performed by molecular dynamics (MD) in vacuo and the solvent effect is calculated a posteriori by solving the Poisson or the Poisson-Boltzmann equation for selected frames of the trajectory. The binding free energy is written as a linear combination of the buried surface upon complexation, SASbur, the electrostatic interaction energy between the ligand and the protein, Eelec, and the difference of the solvation free energies of the complex and the isolated ligand and protein, deltaGsolv. The method uses the buried surface upon complexation to account for the non-polar contribution to the binding free energy because it is less sensitive to the details of the structure than the van der Waals interaction energy. The parameters of the method are developed for a training set of 16 HIV-1 protease-inhibitor complexes of known 3D structure. A correlation coefficient of 0.91 was obtained with an unsigned mean error of 0.8 kcal/mol. When applied to a set of 25 HIV-1 protease-inhibitor complexes of unknown 3D structures, the method provides a satisfactory correlation between the calculated binding free energy and the experimental pIC5o without reparametrization. PMID- 15124935 TI - Selective cyclooxygenase-2 inhibitors: similarities and differences. AB - The enzyme cyclooxygenase (COX) was shown to exist as two distinct isoforms about a decade ago. COX-1 is constitutively expressed as a 'housekeeping' enzyme in nearly all tissues, and mediates physiological responses (e.g. cytoprotection of the stomach, and platelet aggregation). On the other hand, COX-2, expressed by cells involved in inflammation (e.g. macrophages, monocytes, synoviocytes), has emerged as the isoform that is primarily responsible for the synthesis of prostanoids involved in acute and chronic inflammatory states. Consequently, the hypothesis that selective inhibition of COX-2 might have therapeutic actions similar to those of non-steroidal anti-inflammatory drugs, but without causing gastrointestinal side effects, was the rationale for the development of selective inhibitors of the COX-2 isoenzyme. Selective COX-2 inhibitors currently used in the clinic are the sulphonamides celecoxib and valdecoxib (parecoxib is a prodrug of valdecoxib), as well as the methylsulphones rofecoxib and etoricoxib. Furthermore, the phenylacetic acid derivative lumiracoxib has gained permission recently to be marketed in Europe. This review discusses the clinically relevant similarities and differences of these substances, with particular emphasis on their diverse pharmacokinetic characteristics. PMID- 15124936 TI - Regulation of peripheral blood and synovial fluid lymphocyte apoptosis in juvenile idiopathic arthritis. AB - OBJECTIVE: Complex regulatory mechanisms are involved in the induction of apoptosis. Their impairment may play a role in the pathogenesis of several autoimmune diseases. Recently, we have described higher incidences of spontaneous apoptosis of peripheral blood (PB) lymphocytes in children with juvenile idiopathic arthritis (JIA). This study aimed to evaluate the regulatory mechanisms that may be responsible for this phenomenon. METHODS: Thirty-four JIA children were examined and compared with 20 healthy children of similar ages. Expression of regulatory proteins p53, Bax and Bcl-2 in lymphocytes isolated from PB and synovial fluid (SF) was assessed. Serum and SF levels of interleukin-15 (IL-15) were also evaluated. RESULTS: The study showed significantly decreased Bcl-2 expression in JIA PB lymphocytes, compared to both healthy control (p = 0.03) and JIA SF lymphocytes (p = 0.005). There were no significant differences found in Bax expression between groups or compartments examined. However, the Bax/Bcl-2 ratio was nearly two-fold higher in PB lymphocytes than in SF of JIA patients (p = 0.001). p53 expression in PB lymphocytes from both JIA and control children did not statistically differ. In JIA, however, p53 was significantly higher in PB than SF lymphocytes (p = 0.016). IL-15 levels were about 20-fold higher in JIA SF than in serum from either JIA or healthy children (p < 0.0001). CONCLUSION: The results suggest that a higher incidence of apoptosis of PB lymphocytes observed in JIA may be associated with down-regulation of Bcl-2, rather than with changes in expression of Bax and p53. In contrast, the low p53 expression and elevated IL-15 appear to provide mechanisms responsible for suppression of apoptosis in SF cells from JIA patients. PMID- 15124937 TI - Cellular and humoral markers of systemic inflammation in acute reactive arthritis and early rheumatoid arthritis. AB - OBJECTIVE: To compare systemic inflammation in reactive arthritis (ReA), rheumatoid arthritis (RA), and sepsis using novel markers of systemic inflammation, and to study whether they are helpful in distinguishing between ReA and RA. METHODS: In 28 patients with acute ReA, 16 patients with early untreated RA, and 25 patients with blood culture-positive sepsis, phagocyte CD 11b expression was measured by flow cytometry, serum procalcitonin (PCT) levels by immunoluminometric assay, and soluble E-selectin (sE-selectin) levels by enzyme linked immunosorbent assay (ELISA). RESULTS: Neutrophil and monocyte CD11b expression and serum levels of PCT and sE-selectin were higher in patients with sepsis than patients with ReA or RA, or in healthy subjects (all p < 0.01). They were comparable in healthy subjects, ReA, and RA. CONCLUSION: Patients with acute ReA and early RA have normal CD11b expression levels on phagocytes and normal PCT and sE-selectin levels in serum. Elevated levels suggest possible sepsis. PMID- 15124938 TI - Cytochrome p450 1Al gene polymorphisms in patients with psoriatic arthritis. AB - OBJECTIVE: To investigate the associations of cytochrome p450 1A1 (CYP1A1) gene polymorphisms with susceptibility to psoriatic arthritis in Taiwan. METHODS: CYP1A1 gene polymorphisms were determined in 52 patients with psoriatic arthritis and in 90 healthy controls by polymerase chain reaction/restriction fragment length polymorphism. RESULTS: The genotype frequency of CYP1A1 4889A/G was significantly increased in patients with psoriatic arthritis in comparison with healthy controls. The phenotype frequency of CYP1A1 4889G was also significantly increased in patients with psoriatic arthritis. The genotype frequency of CYP1A1 4887C/A was significantly higher in patients with psoriatic arthritis than in controls. The allele and phenotype frequencies of 4887A were also significantly increased in patients with psoriatic arthritis. We also found that the association of CYP1A1 4887A with psoriatic arthritis was independent of 4889G but a synergistic effect was present between CYP1A1 4887A and 4889G. The CYP1A1 4889A/G and 4887C/A polymorphisms were not associated with the manifestations and severity of psoriatic arthritis. CONCLUSION: CYP1A1 4887A and 4889G may be precipitating factors for susceptibility to psoriatic arthritis in Taiwan. An additive effect was found between CYP1A1 4887A and 4889G. PMID- 15124939 TI - HLA-DRB1* alleles and temporomandibular joint erosion in patients with various rheumatic diseases. AB - OBJECTIVE: To investigate the association between HLA antigens and temporomandibular joint (TMJ) erosion, salivary composition, and focal sialadenitis in patients with rheumatic diseases. METHODS: Eighty-four patients, 24 with rheumatoid arthritis (RA), 19 with mixed connective tissue disease (MCTD), 19 with ankylosing spondylitis (AS), and 22 with spondyloarthropathy (SPA) were studied. Each patient underwent clinical examination of the masticatory system, unstimulated and stimulated saliva collection, and minor salivary gland biopsy. Radiographs (OPTG) of the TMJ were obtained, and HLA allele (A, B, C and DRB1*) analysis was performed. Erosion in OPTG was scored from 0 (no erosion) to 4 (condyles totally eroded). In the analysis, scores 0-2 were grouped as normal or mild changes, and scores 3-4 as distinct erosions. One hundred healthy blood donors served as controls for HLA typing. RESULTS: Distinct erosion of the TMJ in OPTG was observed in 22 (27%) patients. It affected four (17%) of the 24 patients with RA, three (17%) of the 18 with MCTD, seven (37%) of the 19 patients with AS and eight (38%) of the 21 with SPA non-significant (NS). The mean erosion scores were 1.7 for RA, 1.3 for MCTD, 2.5 for SPA, and 1.6 for AS patients [probability (p) = 0.04]. The frequency of HLA-B27 antigen was higher in the AS and SPA patients, and that of HLA-DRB1*04 allele higher in RA patients than in control subjects. In the whole patient population, HLA-DRB1*01 allele was significantly associated with erosions 16/36 (44%) versus 6/46 (131%1) (p = 0.0014). In the SPA group, patients with HLA-DRBI*01 allele had a significantly higher occurrence of distinct erosions than patients without this allele [8/10 (80%) versus 0/11 (0%) (p = 0.0002)], whereas DRB1*06 was protective [0/8 (0%) versus 8/13 (62%) (p = 0.018)]. HLA-DRB1*04 was associated with increased salivary IgG in the RA patients. CONCLUSION: HLA antigens are significantly associated with the development of destructive lesions in the TMJ, as well as composition of saliva in patients with various rheumatic diseases. PMID- 15124940 TI - Reactions to disability in patients with early versus established rheumatoid arthritis. AB - OBJECTIVES: This study examined differences in reactions to disability between early and established rheumatoid arthritis (RA) patients, and whether these reactions were related to age, physical functioning, acceptance of illness, or self-efficacy. METHODS: Thirty-four patients with early RA (< 2 years since diagnosis) and 84 patients with established RA (> 4 years since diagnosis) completed the Reactions to Impairment and Disability Inventory (RIDI), and measures of anxiety, depression, acceptance of illness, self-efficacy, and physical functioning. RESULTS: Early RA patients reported greater future denial than established RA patients. Younger patients reported more hostility than older patients. Accepting the illness was uniquely related to less anger and hostility. Higher self-efficacy for pain specifically related to greater shock, while patients with poorer self-efficacy for other symptoms reported worse anxiety, depression, shock, and anger. CONCLUSIONS: Denial may be a coping strategy in the early stages of RA: anxiety, depression, shock, and anger appear to persist. Longitudinal studies of RA patients from diagnosis are required to plan interventions timed to maximize patient benefit and optimize healthcare resource utilization. PMID- 15124941 TI - Estimating the prevalence among Caucasian women of primary Sjogren's syndrome in two general practices in Birmingham, UK. AB - OBJECTIVE: To establish the prevalence among women of primary Sjogren's syndrome (PSS) in Birmingham, UK. METHODS: Eight hundred and forty-six female Caucasians from two general practitioner lists were invited to complete a questionnaire that included a screening question on dry eyes and mouth. Individuals who responded positively were evaluated further. RESULTS: Overall, 65/% of individuals who were sent a questionnaire responded. Two had possible PSS, but were negative for anti Ro/La antibodies. Our estimates of the prevalence of PSS ranged from < 0.1% up to 0.4%, depending on the assumptions used. CONCLUSION: Our data support previous studies suggesting a prevalence of PSS in the community of 0.1-0.6% rather than those suggesting a higher figure. PMID- 15124942 TI - Possible importance of immunoglobulin E in foetal loss by mothers with anti-SSA antibody. AB - OBJECTIVE: The incidence of foetal loss and/or adverse foetal outcomes, including congenital heart block (CHB), has been investigated in mothers with anti-SSA antibody detected by immunodiffusion or counter-immunoelectrophoresis methods. We investigated the relationship between several serum parameters (such as autoantibodies and immunoglobulins) and foetal loss in patients with anti-SSA antibody, measured by enzyme-linked immunosorbent assay (ELISA). MATERIAL AND METHODS: Thirty-seven women who showed positivity for anti-SSA antibody and had a history of pregnancy were included in this study. Immunoglobulins and several autoantibodies were assayed by routine laboratory methods at our hospital. RESULTS: Our data indicated that immunoglobulin E (IgE) levels were significantly higher in the anti-SSA antibody positive women with foetal loss than in those without, and that a strong positive correlation between IgE and anti-SSA antibody levels was observed in the former group, but not in the latter. CONCLUSION: The serum IgE level seems to be an important factor in the occurrence of foetal loss in mothers with anti-SSA antibody detected by ELISA. PMID- 15124943 TI - Teaching medical students musculoskeletal examination skills: identifying barriers to learning and ways of overcoming them. AB - OBJECTIVES: To elicit the barriers to the effective teaching of musculoskeletal examination skills amongst medical students. METHODS: This was a qualitative study including six focus groups with specialities most often involved in delivering musculoskeletal clinical teaching: rheumatology, orthopaedics, general practice, and geriatrics. RESULTS: The main barriers to the delivery of effective clinical teaching included the lack of agreement on what to teach, lack of confidence in teaching amongst non-musculoskeletal specialities, and poor communication between specialities. CONCLUSIONS: There is a need to overcome the identified barriers if musculoskeletal clinical teaching to medical students is to be improved. In particular, there is a need to agree which examination skills medical students should learn. PMID- 15124944 TI - Bilateral pulmonary artery aneurysms with protein C and protein S deficiency in a patient with Behcet's disease. AB - Behcet's disease is a chronic inflammatory disorder of unknown aetiology, characterized by recurrent attacks. Pulmonary artery aneurysm is a rare but serious complication of Behcet's disease. We describe a patient with Behcet's disease and protein C and S deficiency who developed bilateral pulmonary artery aneurysms. PMID- 15124945 TI - Acute anterior uveitis in association with an outbreak of Campylobacter jejuni infection. AB - We report a case of acute anterior uveitis (AAU) in association with an outbreak of Campylobacter jejuni infection, with the first estimation of the incidence of AAU triggered by Campylobacter, and discuss reactive ophthalmological complications (AAU, iritis, and conjunctivitis) attributable to Campylobacter. PMID- 15124946 TI - Systemic lupus erythematosus associated with cytomegalovirus infection. PMID- 15124947 TI - Scleroderma in a patient with X-linked agammaglobulinaemia. PMID- 15124948 TI - Exacerbation of cryoglobulinemia associated with intravenous radiographic contrast medium. PMID- 15124949 TI - Acid transport through gastric mucus. AB - The gastric mucosa is frequently exposed to endogenously secreted hydrochloric acid of high acidity. Gastric mucosal defense mechanisms are arranged at different levels of the gastric mucosa and must work in unison to maintain its integrity. In this work, several mechanisms underlying gastric mucosal resistance to strong acid were investigated in anesthetized rats and mice. The main findings were as follows: Only when acid secretion occurred did the pH gradient in the mucus gel withstand back-diffusion of luminal acid (100 mM or 155 mM HCl), and keep the juxtamucosal pH (pH(jm)) neutral. Thus, with no on-going acid secretion and low luminal pH, the pH gradient was destroyed. Bicarbonate ions, produced concomitant with hydrogen ions in the parietal cells during acid secretion and transported by the blood to the surface epithelium, were carried transepithelially through a DIDS-sensitive transport. Prostaglandin-dependent bicarbonate secretion seemed to be less important in maintaining a neutral pH(jm). Removal of the loosely adherent mucus layer did not influence the maintenance of the pH(jm). Hence, only the firmly adherent mucus gel layer, approximately 80 microm thick, seemed to be important for the pH(jm). Staining of the mucus gel with a pH-sensitive dye revealed that secreted acid penetrated the mucus gel from the crypt openings toward the gastric lumen only in restricted paths (channels). One crypt opening was attached to one channel, and the channel was irreversibly formed during acid secretion. Gastric mucosal blood flow increased on application of strong luminal acid (155 mM HCl). This acid-induced hyperemia involved the inducible but not the neural isoform of nitric oxide synthase. These results suggest a novel role for iNOS in gastric mucosal protection and indicate that iNOS is constitutively expressed in the gastric mucosa. PMID- 15124950 TI - Bacteriuria in spinal cord injured patients with neurogenic bladder dysfunction. AB - The occurrence of bacteriuria in spinal cord injured patients with neurogenic bladder dysfunction who used clean intermittent catheterisation to empty their bladders was studied in order to examine cut-off concentration breakpoints for significant bacteriuria in this group of patients using procedures of the European Urinanalysis Guideline. 344 samples were cultured, yielding 285 isolates. Coagulase-negative staphylococci (27%), Enterococci (25%), Klebsiella spp (19%), and Escherichia coli (12%) were the most common findings. Bacteria grew at concentrations of 10(5)-10(8) cfu/L, but only a few at 10(4) cfu/L. It is concluded that low bacterial concentrations in the urine (10(5) cfu/L) of patients with neurogenic bladder dysfunction who are on intermittent catheterisation might be as significant for bladder contamination with bacteria as a high bacterial concentration and can possibly be responsible for bladder infections. PMID- 15124951 TI - The influence of sociodemographic characteristics on health care utilisation in a Swedish municipality. AB - During recent years there has been a debate on factors that might influence health care utilisation, and the possibilities to predict such utilisation in order to facilitate the allocation of health care resources in the community. Currently only the age distribution in the population is usually used for such predictions. In this study the influence of a number of other sociodemographic characteristics on health care utilisation, in addition to age and sex, was assess in a small Swedish community. A postal questionnaire, containing questions on health care utilisation and a number of sociodemographic factors, was sent to a random, stratified sample of the population of Habo municipality, Sweden. Out of the sampled 1312 subjects, 827 (63%) responded to the questionnaire. Subjects aged 65 years and above had higher odds for having consulted a physician, having been hospitalised and having a cost limitation card compared to those below age 65. Men had significantly lower odds than women for having consulted a physician, having visited other health care providers and having a cost limitation card. Singles, as compared to married subjects, had lower odds for all health care measures except having consulted a physician. Subjects with sick leave or disability pension, as compared to working subjects, had higher odds for most health care utilisation measures. A number of sociodemographic factors were thus associated with health care utilisation and might be considered in the health care resources allocation process. PMID- 15124952 TI - Effects of haemodialysis and continuous ambulatory peritoneal dialysis on P300 cognitive potentials in uraemic patients. AB - The aim of this study was to determine the effects of haemodialysis (HD) and continuous ambulatory peritoneal dialysis (CAPD) on P300 cognitive potentials in patients with chronic renal failure (CRF) and to find out if P300 potential is a valuable marker for following subclinical cognitive disorder. This study was performed in 42 patients with chronic uraemia, of whom 25 were on HD and 17 on CAPD, and in 25 healthy subjects. All the subjects were investigated in terms of P300 cognitive potential obtained from auditory stimuli with the oddball paradigm and the Mini-Mental State (MMS) examination. Patients undergoing HD were evaluated before (pre-dialysis) and after (post-dialysis) standard HD treatment. P300 latency was longer in HD patients than in the control group and CAPD patients. The MMS score was greater and P300 latency was shorter after a standard HD session when compared to pre-dialysis values in HD patients (p < 0.001 for both values). There was a negative correlation between P300 latency and MMS scores, and a positive correlation between P300 amplitude and MMS scores. In conclusion, P300 is useful for evaluating cognitive function in uraemia, even in asymptomatic patients, and CAPD is superior to HD in the management of cognitive impairment. PMID- 15124953 TI - Super bone scan due to bone marrow metastases appearing 19 years after surgery for early gastric cancer--a case report. AB - A 54 year old man presenting a so-called super bone scan is reported. The patient had a past history of subtotal gastrectomy due to early gastric cancer 19 years previously. Laboratory data indicated microcytic anemia and high serum alkaline phosphatase. Both the serum calcium and phosphate levels were within normal ranges. In spite of extensive examination for a primary malignant lesion in the organs including the remaining stomach, no solid tumors were identified. Pathological examinations revealed that the tumor cells in the biopsy specimen from the lumbar spine were almost identical to those in the gastric cancer excised 19 years previously. We considered that micrometastases in the bone marrow existed at the time of the initial surgery for gastric cancer and reappeared 19 years later showing super bone scan. PMID- 15124954 TI - The MRI features and treatment of scapulothoracic bursitis: report of four cases. AB - Among cases of soft tissue tumours arising between the inferior angle of the scapula and thoracic wall, scapulothoracic bursitis has rarely been reported. All the reported cases were surgically treated and there have been no reports about the results of conservative treatment observed for a long period. Therefore, we retrospectively evaluated the clinical findings, magnetic resonance imaging (MRI) features, and treatment procedures in order to elucidate the differential diagnosis for, and efficacy of, conservative treatment. The patients were those who consulted our department of orthopedic surgery from 1994 to 1998 with complaints of thoracic back tumours. The patients were 2 men and 2 women whose ages ranged from 46 to 66 years. The tumour locations, MRI findings and treatments were evaluated. The cysts appeared in the inferior angle of the scapula with the shoulder flexed and adducted. There was neither pain nor tenderness, local heat nor redness. The cysts were elastic hard, fluctuating, and less mobile and the margins were well circumscribed by palpation. The diameters were from 6 x 6 cm to 20 x 15 cm. On MR imaging, the cysts were located between the serratus anterior muscle and the chest wall. Slightly high signal intensity on T1WI, high on T2WI and fluid-fluid levels were seen in the cysts in three cases. For the treatment, in one case we performed multiple aspirations until a reduction appeared to occur. The average aspirated contents of the cyst were 100 mL to 200 mL and were bloody-serous. In two cases no special treatments were employed, and in one case the tumour was surgically removed because it was unresponsive to aspiration. In the three cases with conservative treatment, the follow-up time was 8, 9 and 25 months, respectively. The tumours had disappeared in all cases at the time of the final follow-up. High signal intensities on T1 and T2 weighted images, and fluid-fluid level on T2 weighted images were very useful MRI features for the diagnosis of scapulothoracic bursitis. Our observations indicate that conservative treatment can be successful and surgery is not necessary except for cases with pain, excessive friction, or dysfunction. PMID- 15124955 TI - Periosteal chondroma of the fifth toe--a case report. AB - A case of a periosteal chondroma arising in the fifth toe is reported. A 19-year old man presented a palpable mass of his left fifth toe with pain for 2 years. Plain radiographs showed mild irregularity of the cortical bone of the fifth proximal phalanx. Magnetic resonance imaging revealed a tumour attached to the fifth proximal phalanx with low signal intensity on T1-weighted images and high signal intensity on T2-weighted images without intramedullary extension. The tumour was enhanced heterogeneously after intravenous injection of gadolinium diethylene triamine penta-acetic (GD-DTPA). The tumour was removed, together with the covering periosteum. It was diagnosed as periosteal chondroma histologically. PMID- 15124956 TI - Through the eyes of Hollywood: images of social workers in film. AB - This article reports on research that investigated the images of social workers as portrayed in movies as a major medium of popular culture. Findings from an analysis of 44 movies spanning the period from 1938 to 1998 are presented with particular attention to the themes of gender, race, and class. Using the concept of motivated representations in popular culture, we suggest that these popular constructions of social workers are perhaps motivated by efforts to maintain the status quo of the dominant society, rather than to promote social change and social justice. PMID- 15124957 TI - The public's perception of social work: is it what we think it is? AB - This article reports on the public's current perception of the social work profession as examined in a nationally representative, random digit telephone survey of 386 people. The survey asked respondents about their knowledge, beliefs, and attitudes regarding the profession and its practitioners and required respondents to make comparisons between social workers and other helping professionals such as psychiatrists, psychologists, counselors, nurses, and clergy. Results of the survey indicated that for the most part, a majority of the public understands the social work profession and in many ways recognizes its value. PMID- 15124958 TI - Personal, family, and multiple barriers of long-term welfare recipients. AB - This article reports the results of an in-depth, descriptive study of long-term welfare recipients. A random sample of 284 respondents, all of whom had received public assistance for at least 36 months, were interviewed. Results illustrate personal barriers to self-sufficiency, including physical health problems that prevent work, severe domestic violence, educational deficits, substance abuse, learning disabilities, child behavior problems, generalized anxiety disorder, posttraumatic stress disorder, and clinical depression. Study findings underscore the complex, persistent, and multiple difficulties experienced by this population and suggest that for some welfare recipients, long-term (even lifelong) financial supports and social services may be in order. Implications for policy and programming are discussed. PMID- 15124959 TI - Difficulties after leaving TANF: inner-city women talk about reasons for returning to welfare. AB - People who leave welfare commonly return, and this phenomenon has become more pressing in the time-limited TANF program. Fostering stable TANF exits may be particularly difficult in poor inner-city areas because of job shortages and neighborhood deterioration. Perspectives of TANF leavers from five focus groups in Chicago about problems leading to welfare returns are presented. Participants indicated that low wages and unstable jobs were most often responsible for TANF returns. Obtaining health care and child care and inconsistent performance of TANF caseworkers were stressed as contributing factors. Participants also focused on strengths helpful in sustaining exits from TANF, particularly psychological benefits associated with working and informal supports received from family members and friends. The implications of recipient-identified problems and strengths are discussed, including balancing "work first" employment policies with substantive educational and job development policies. Strategies for improving the performance of TANF case planning are discussed. PMID- 15124960 TI - Low-income young fathers: contexts, connections, and self. AB - Although teenage pregnancy is at the center of much current social concern and political debate, the focus tends to be on the young mothers and their children. The lives and parenting experiences of young fathers typically receive less attention from researchers, practitioners, and policymakers. This article presents findings from a qualitative research study of 25 low-income young fathers. Young men were asked questions about their own life experiences and social contexts, their connections with their children and female partners, and the implications these had for sense of self. They were interviewed again one year later. The majority of young fathers were found to be involved significantly in the lives of their children, despite their own struggles. This in turn helped them feel positive about their sense of self. Implications for social policy and programs are discussed. PMID- 15124961 TI - Presence of social workers in nongovernment organizations. AB - This study investigated the job categories full-time social workers occupy in 20 international nongovernment organizations (NGOs). Social workers holding BSW or higher degrees account for 95 percent of the program director and coordinator positions, indicating that now there may be greater use of these professionals. But non-social workers filled 83 percent of the direct services positions. Also disconcerting was the small representation of social workers in administration and development. Thus, social workers are used in program coordination, but underused in leadership and services provision. Organizational resources and programs, when linked to larger annual budgets, may indicate an NGO's ability to provide specialized direct services using social work expertise. PMID- 15124962 TI - Whistle-blowing as a form of advocacy: guidelines for the practitioner and organization. AB - Advocacy has been an inherent component of social work since the mid-1800s. The NASW Code of Ethics explicitly promotes advocacy as an ethical stance against inhumane conditions. Whistle-blowing, on the other hand, occurs mostly in the business and public administration disciplines and is relatively unknown in the social work profession. Using facts from composite cases of whistle-blowing incidents, the purpose of this article is to review the social work profession's current stance on advocacy to protect clients' rights, define and describe theoretical and practical knowledge about whistle-blowing based on a literature review, explain whistle-blowing as a special form of advocacy, and offer guidance to potential whistle-blowers and their organizations on how to handle situations in which whistle-blowing is likely to be considered an option. PMID- 15124963 TI - A qualitative investigation of adherence issues for men who are HIV positive. AB - People who are HIV-positive and people who have been diagnosed with AIDS have new hope in the form of antiretroviral medications. However, many social workers remain unaware of the rigors of this treatment regimen and the difficulties it presents. This article presents the results of a qualitative study of HIV positive individuals who have struggled with the adherence issues endemic to antiretroviral treatment and seeks to give voice to their experiences. The article seeks to educate social workers about antiretroviral treatment for HIV, importance of adherence, and intervention strategies. PMID- 15124964 TI - Relational spirituality and social caregiving. AB - Relational spirituality suggests that strengthening one's relatedness to other people and to sources of meaning beyond self-interest produces joy and vitality, which sustain the capacity to care. This perspective informs a qualitative study of 50 female services providers and social reformers. The interviews revealed that the women's caregiving was sustained not only by efficacy or success, but also by connections to their work, to their social networks, to their clients, and to a sacred source of love and strength. Implications from the study focus on strategies to strengthen caregivers' sense of relatedness to sustain the capacity for social caregiving. PMID- 15124965 TI - Development of innovative group work practice using the intervention research paradigm. AB - Rothman and Thomas' intervention research (IR) paradigm provides an alternative, developmental research method that is appropriate for practice research, especially at the early stages. It is more flexible than conventional experimental designs, capitalizes on the availability of small samples, accommodates the dynamism and variation in practice conditions and diverse populations, and explicitly values practitioners' insights. This article describes two studies to exemplify how the features of IR methodology are well suited to research on innovative group interventions. In one study, a face-to face psychoeducational group for patients living with sickle cell anemia and chronic depression was used. The other study used an online job stress support group for social workers. The challenges and opportunities that can arise when applying the IR paradigm to develop new interventions are also discussed. PMID- 15124966 TI - Who we are, where we come from, and some of our perceptions: comparison of social workers and the general population. AB - Essentially nothing is known about the national population of social workers. Accordingly, to enhance our understanding of the individuals who make up the social work profession, this study used nationally representative data to compare the characteristics of graduate- and bachelor's-level workers with those of the general population. Data are provided on present and childhood demographics as well as a number of current perceptions. In terms of perceptions, social workers were more likely to rate their lives as exciting, to live in racially mixed neighborhoods, to be stronger proponents of controversial speech, and to be perceived by survey interviewers as friendlier than the general public. A number of suggestions for further research are provided to flesh out the initial findings presented in this article. PMID- 15124968 TI - When a social worker becomes a voluntary commissioner and calls on the Code of Ethics. AB - When practicing in a new arena, where does a social worker turn for guidance? This article discusses the author's experiences as chair of a governor-appointed Special Commission to Study Sexual Orientation Discrimination. The Code of Ethics of the National Association of Social Workers proved a useful guide in framing the social work role and in helping to resolve thorny issues that emerged. PMID- 15124967 TI - Is Christian religious conservatism compatible with the liberal social welfare state? AB - This article examines the rise of Christian religious conservatism and explores whether the theological views of the conservative Christian movement are compatible with the liberal social welfare state. The authors conclude that the driving force behind social change should remain with the state, even though faith-based initiatives can provide some basic supports that the social welfare state can use. PMID- 15124969 TI - Social justice and the global economy: new challenges for social work in the 21st century. AB - The globalization of the economy creates new challenges for social work in the arenas of social and economic justice. This article outlines social justice issues related to the debt crisis of the Global South and sweatshops. A presentation of colonial precursors is followed by a detailed examination of these global institutions with an emphasis on the vulnerability, disempowered status, and exploitation of poor people of the Global South. Connections with global inequities in wealth, income, and the distribution of resources are made explicit. The article explores domestic social justice problems as possible points of connection with these issues. Finally, the authors give recommendations for social work education, advocacy, and activism. PMID- 15124970 TI - Increasing the cultural responsiveness of family group conferencing. AB - Child welfare struggles to manage child abuse and neglect and to seek permanency for children, while being culturally responsive to the communities it serves. Family group conferencing, piloted in New Zealand and now used in the United States and other countries, is a strengths-based model that brings together families and their support systems to develop and carry out a plan that protects, nurtures, and safeguards children and other family members. This article describes the model and a culturally competent method for assessing and adapting the model for the African American, Cherokee, and Latino/Hispanic communities in North Carolina. PMID- 15124971 TI - Barriers to family-centered services for infants and toddlers with developmental delays. AB - Social workers have become increasingly involved in providing family-centered services to families with infants and toddlers who have developmental delays. This study examined the perceptions of barriers to family-centered services of 22 family members and 20 services providers in one local services delivery system. A family-centered approach focuses on helping families cope with the challenges of having children with developmental delays, helping families work collaboratively with services providers, and supporting families as they make decisions about services. Families have experienced barriers related to aspects of the traditional Part C early intervention system and to providers outside the realm of traditional early intervention services and barriers related to individual personality characteristics. Participants suggested several strategies for empowering families to take a more active role on early intervention teams. PMID- 15124972 TI - Gender-based salary inequity in social work: mediators of gender's effect on salary. AB - This study examined the direct effect of gender, controlling for years of experience, job role, and other variables, in a sample of 359 Pennsylvania social workers. Men social workers received significantly more yearly salary (an estimated 3,665 dollars more) than women social workers. A path analysis suggested that the salary advantage for men that was attributable to their acquiring more experience and management positions was slightly larger than the direct effect of gender. The ability of merit variables to explain salary was somewhat greater for men than women. Other variables that mediated the effect of gender on salary were MSW specialization, working in a social work or a related area, and practice area (children and youths, health, mental health, or other area). The results suggest that substantial gender disparity continues to exist in social worker salaries. PMID- 15124973 TI - Social work is standing on the legacy of Jane Addams: but are we sitting on the sidelines? PMID- 15124974 TI - Setting the record straight: social work is not a female-dominated profession. PMID- 15124975 TI - Social work and the Supreme Court: a clash of values; a time for action. PMID- 15124976 TI - Television and the public image of social workers: portrayal or betrayal? PMID- 15124977 TI - Drugs used in the treatment of opioid tolerance and physical dependence: a review. AB - Opioid drugs used in the treatment of severe pain are known to produce tolerance that requires a dose increase to maintain a sufficient analgesic effect. As this is connected with side effects such as respiratory depression, it is highly desirable to avoid or at least attenuate the development of tolerance. Closely related, but in some respect dissociable, is the phenomenon of physical dependence, which becomes apparent particularly in heroin withdrawal. Our knowledge about the mechanisms underlying tolerance has increased dramatically in recent years, but a final picture of the importance of each particular mechanism under in vivo conditions has not yet emerged. Recent studies suggest that the so called receptor down-regulation is not the main mechanism in vivo. A desensitization on the basis of receptor decoupling, receptor internalization and increased alternative coupling to stimulatory G-proteins have been demonstrated. However, a functional antagonism of the opioid effects seems to be clinically most important, mediated by the activation of NMDA receptors, up-regulation of adenylyl cyclase and nitric oxide synthase. Drugs blocking these mechanisms are the most promising option in the treatment of tolerance. Namely, alpha2 adrenoreceptor agonists such as clonidine and NMDA antagonists such as ketamine or dextromethorphan have been used to minimize tolerance development during opioid treatment. Moreover, clinical strategies such as opioid rotation and multimodal analgesia, i.e. the simultaneous application of several analgetics of different type, have proven to be successful approaches. PMID- 15124978 TI - Hemodynamic effects of levosimendan in patients with low-output heart failure after cardiac surgery. AB - Following cardiac surgery, low-output syndrome is relatively common. Since this condition can lead to serious consequences, this postsurgical, low-output state should be reversed whenever possible. Patients with low-output syndrome need drug and fluid management aimed at enhancing cardiac contractility and at facilitating optimal myocardial loading. The objective of this pilot study was to evaluate whether benefits of levosimendan, a new calcium-sensitizing agent approved for treatment of patients with acute exacerbation of chronic heart failure, could be extended to patients with low-output syndrome following cardiac surgery. For this study, each patient was given levosimendan as a loading dose of 12 microg/kg over 10 minutes, followed by a continuous infusion of 0.1 microg/kg/min for 12 hours. Of 11 postsurgical patients with severely impaired cardiac output and hemodynamic compromise, 8 patients (73%) showed evidence of combined hemodynamic improvement (> 30% increase in cardiac index and PCWP corrected to < 18 mmHg) within 3 h after the start of levosimendan infusion. Specifically, cardiac index and stroke volume were significantly increased, while mean arterial pressure, indexed systemic vascular resistance, mean pulmonary pressure, right arterial pressure, and pulmonary capillary wedge pressure were all significantly lowered. Taken together, such changes showed enhanced cardiac output along with significantly decreased preload and afterload--conditions associated with recovery of cardiac function. Levosimendan is thus highly favorable for short-term treatment of patients with low cardiac output following cardiac surgery. PMID- 15124979 TI - Effects of micronized fenofibrate on insulin resistance in patients with metabolic syndrome. AB - BACKGROUND AND OBJECTIVE: Metabolic syndrome is characterized by insulin resistance (IR) as well as dyslipidemia, ventral overweight, hypertension and elevated fasting plasma glucose. Since diabetic and prediabetic states are commonly associated with hypertriglyceridemia, fenofibrates have been used in such patients. The aim of this pilot open trial was to study the influence of micronized fenofibrate on insulin resistance and plasma insulin levels in prediabetic and diabetic patients. SUBJECTS: From 114 dyslipidemic patients, 31 with dyslipidemia and insulin resistance were selected to take part in the study. Of the 31 patients, 20 were nondiabetic and only 11 had noninsulin-dependent diabetes mellitus. Eighteen dyslipidemic patients acted as controls. METHODS: Insulin resistance was assessed in a short-term insulin tolerance test. Plasma insulin, antiinsulin antibodies, lipid parameters and the insulin sensitivity index (ISI) were measured at entry and after a 3-month therapy with 200 mg micronized fenofibrate daily. RESULTS: Three-month therapy with micronized fenofibrate resulted in significant ISI increase and was accompanied by a decrease in plasma insulin levels in dyslipidemic patients with metabolic syndrome. ISI also improved in patients with type 2 diabetes mellitus and there was an unexpected increase in plasma insulin levels. Antiinsulin antibodies were unchanged throughout the trial. Reductions in plasma triglycerides and total cholesterol exceeding 50% and 20%, respectively, were observed in patients with metabolic syndrome. These changes were accompanied by an increase in mean levels of plasma high-density lipoprotein (HDL) cholesterol (above 35%). CONCLUSIONS: Micronized fenofibrate is an effective drug in normalizing lipid-lipoprotein levels in patients with metabolic syndrome. After a 3-month fenofibrate therapy, insulin resistance was reduced in a group of patients with dyslipidemia and metabolic syndrome. PMID- 15124980 TI - Influence of an antivertiginous combination preparation of cinnarizine and dimenhydrinate on event-related potentials, reaction time and psychomotor performance--a randomized, double-blind, 3-way crossover study in healthy volunteers. AB - In the present comparative, double-blind, 3-way crossover study, possible effects of an antivertiginous combination preparation on event-related potentials (ERPs) and performance were investigated. Twenty-one healthy volunteers received 4 doses (within 24 h) of a fixed combination of cinnarizine 20 mg and dimenhydrinate 40 mg (Arlevert, ARL), dimenhydrinate 50 mg, or a placebo, in randomized order at 1 week intervals. Auditory event-related potentials (ERPs), reaction time (RT) and psychometric tests were assessed before as well as 60 and 150 minutes after the intake of the 1st (Day 1) and the 4th (Day 2) dose of study medication. The evaluation was primarily based on the difference in the outcomes measured 150 min after the 4th dose (t5) and those before the start of medication intake (t0). None of the medications affected the latency and amplitude of the sensory ERP component N100, neither under passive listening nor under discrimination task conditions. The latency of P300 in response to the rare target tones (oddball paradigm and binary series), showed significant (p < 0.05) delays after 4 doses of dimenhydrinate (18-24 ms), and no significant differences between ARL (3-17 ms) and either dimenhydrinate or placebo (4-13 ms). Responses to nontarget tones remained almost unaffected after medication intake. The secondary analysis of the P300 amplitude showed the greatest decreases under DH in both active series, with no significant differences between ARL and either DH or placebo. The 3 medications did not significantly prolong RT nor did they impair the performance of psychometric tests, or cause significant shifts of current mood. The combination preparation ARL showed the lowest rate of adverse events (n = 1), followed by dimenhydrinate (n = 3) and placebo (n = 6). Two subjects withdrew because of adverse events, both after the intake of placebo. In conclusion, the results gave no evidence for an impairment of central information processing and psychomotor performance after multiple dosing with the fixed combination ARL in healthy volunteers, which might, when present, represent an adverse reaction limiting its use in antivertiginous therapy. No significant differences were found between ARL and placebo. PMID- 15124981 TI - Pharmacokinetic model of R-roscovitine and its metabolite in healthy male subjects. AB - OBJECTIVE: To characterize the pharmacokinetics of R-roscovitine, a novel cyclin dependent kinase inhibitor, and its carboxylate metabolite in man. METHOD: Twelve healthy male subjects received single oral doses of 50, 100, 200, 400 or 800 mg in a hierarchical 3-period, 6-sequence crossover design. One dose was given after breakfast, the others under fasting conditions. R-roscovitine and the carboxylate metabolite were measured in plasma and urine. A 2-compartment model for R roscovitine with 1 compartment for the metabolite and a component for first-pass extraction was adequate. Protein binding was calculated from plasma and urine data. RESULTS: R-roscovitine undergoes nonsaturatable first-pass extraction, rapid metabolism, exhibits high nonsaturated protein binding, is slowly absorbed from the GI tract and is rapidly and extensively distributed into tissues. The slow release of the molecule from tissue determines the apparent terminal half life. Food delays the absorption and slows down the absorption rate but does not influence bioavailability. The formation rate of the carboxylate is a determinant of the plasma concentrations of this metabolite. It has low protein binding, limited tissue distribution and a renal clearance reflecting with good water solubility. CONCLUSION: The compartmental analysis clarified important pharmacokinetic aspects relevant for the clinical development of the compound. PMID- 15124982 TI - A comparison of two sparse sampling population pharmacokinetic approaches for the estimation of pharmacokinetic parameters in children. AB - BACKGROUND AND OBJECTIVE: The objectives of this study were to assess pharmacokinetic parameters (clearance, volume and half-life) in children using sparse sampling population as well as Bayesian (post hoc) approach. METHODS: Three drugs were selected for this study. Two sparse sampling methods (variable or fixed) using population and Bayesian approaches were used to assess pharmacokinetic parameters in children following a single oral dose. The initial estimates of the model parameters and inter- and intrasubject variability were obtained from the pharmacokinetic studies conducted in adults. The estimated pharmacokinetic parameters using sparse sampling (3 blood samples) were compared with the pharmacokinetic parameters obtained by extensive sampling (> or = 7 blood samples). RESULTS AND CONCLUSIONS: The results indicated that both variable and fixed sampling approaches could be used to estimate mean population as well as individual pharmacokinetic parameters in children with fair degree of accuracy. The methods described here can be used to assess either population or individual pharmacokinetic parameters in children, provided there is a prior knowledge of the pharmacokinetics of a drug in adult population. PMID- 15124983 TI - Patterns of systemic antibiotic use in a tertiary hospital in Israel in the years 1998-2000. AB - OBJECTIVES: To investigate the pattern of antibiotic use in the hospital over a 3 year period according to individual drugs and hospital departments. SETTING: 335 adult beds of a tertiary hospital in Northern Israel during the years 1998-2000. An antibiotic control policy restricts the use of the most expensive antibiotics and those with broad spectrum of activity and a major impact on bacterial resistance. METHODS: The ATC/DDD and DU 90% methodologies were used. The use of antibiotics was expressed as the number of defined daily doses (DDD) per 100 bed days. RESULTS: The total antibiotic use varied during the study period from 93.7 101.0 DDD/100 bed-days (p < 0.1). Thirteen drugs accounted for 90% of the total volume. The use of broad spectrum penicillins was the highest of all drugs followed by cephalosporins and oral quinolones. The highest rates of antibiotic use were found in the departments of ENT, urology, gynecology and orthopedics and in the intensive care unit (ICU). The total restricted antibiotics use was 7.2 DDD/100 bed-days and was the highest in the ICU. CONCLUSIONS: The ATC/DDD methodology provided delineation and interpretation of antibiotic usage patterns in the hospital. Although the overall use is higher then that found in several reports from European hospitals, stratification by individual drugs and by hospital department yielded similar trends. PMID- 15124984 TI - The current NRC definitions of therapy misadministration are vague, do not reflect the norms of clinical practice, and should be rewritten. For the proposition. PMID- 15124985 TI - The current NRC definitions of therapy misadministration are vague, do not reflect the norms of clinical practice, and should be rewritten. Against the proposition. PMID- 15124986 TI - A new MLC segmentation algorithm/software for step-and-shoot IMRT delivery. AB - We present a new MLC segmentation algorithm/software for step-and-shoot IMRT delivery. Our aim in this work is to shorten the treatment time by minimizing the number of segments. Our new segmentation algorithm, called SLS (an abbreviation for static leaf sequencing), is based on graph algorithmic techniques in computer science. It takes advantage of the geometry of intensity maps. In our SLS approach, intensity maps are viewed as three-dimensional (3-D) "mountains" made of unit-sized "cubes." Such a 3-D "mountain" is first partitioned into special structured submountains using a new mixed partitioning scheme. Then the optimal leaf sequences for each submountain are computed by either a shortest-path algorithm or a maximum-flow algorithm based on graph models. The computations of SLS take only a few minutes. Our comparison studies of SLS with CORVUS (both the 4.0 and 5.0 versions) and with the Xia and Verhey segmentation methods on Elekta Linac systems showed substantial improvements. For instance, for a pancreatic case, SLS used only one-fifth of the number of segments required by CORVUS 4.0 to create the same intensity maps, and the SLS sequences took only 25 min to deliver on an Elekta SL 20 Linac system in contrast to the 72 min for the CORVUS 4.0 sequences (a three-fold improvement). To verify the accuracy of our new leaf sequences, we conducted film and ion-chamber measurements on phantom. The results showed that both the intensity distributions as well as dose distributions of the SLS delivery match well with those of CORVUS delivery. SLS can also be extended to other types of Linac systems. PMID- 15124987 TI - Determination of dose-area product from panoramic radiography using a pencil ionization chamber: normalized data for the estimation of patient effective and organ doses. AB - The aims of the present study were (a) to investigate the potential of pencil ionization chamber to be used for the determination of dose-width product (DWP) and dose-area product (DAP) from panoramic radiographic exposures and (b) to provide data normalized to DAP for the determination of patient effective and gonadal dose from panoramic radiography performed in any laboratory. A pencil ionization chamber commonly used to measure CT dose index (CTDI) in CT scanners was employed to determine DWP for various combinations of panoramic exposure settings at the beam exit slit of a Cranex Tome panoramic x-ray unit (Soredex, Helsinki, Finland). DWP values were also measured using an array of thermoluminescence dosimeters. Reproducibility of the DWP measurement was tested. The effect of milliamperage and kilovoltage of panoramic exposures on DWP was investigated. DAP was estimated using the value of DWP measured using the pencil ionization chamber and the beam exit slit length measured using dosimetric film attached on the beam exit slit. A Rando anthropomorphic phantom appropriately loaded with thermoluminescent dosimeters (TLDs) was used to obtain organ dose and effective dose values from panoramic radiography. Reproducibility of DWP determination using the proposed method was better than 1.5%. DWP was found to be linearly related to milliamperage (r>0.999, p<0.001) and to kilovoltage raised in a power ranging from 2.18 to 2.55. DWP measured using the pencil chamber was found to be up to 11% higher than the corresponding values determined using TLD array. The panoramic exposure obtained with settings appropriate for the typical adult patient was found to result in 0.008 mSv patient effective dose, 0.0002 mGy gonadal dose, and 11.3 cGy cm2 DAP. The use of a pencil ionization chamber is proposed for the determination of DWP and DAP from panoramic radiographic exposures. Normalized data over DAP were provided for the determination of patient effective and gonadal dose from panoramic radiography. PMID- 15124988 TI - Noise in flat-panel displays with subpixel structure. AB - Subpixel structures found in medical monochrome active-matrix liquid crystal displays (AMLCDs) affect noise estimates measured with conventional methods. In this work, we discuss methods that identify sources of noise and permit the comparison of luminance noise estimates across technologies independent of pixel design and device technology. We used a three-million pixel AMLCD with a pixel structure consisting of three color stripes, each in a two-domain, in-plane switching mode. Images of uniform fields displayed on the AMLCD were acquired using a low-noise, high-resolution CCD camera. The camera noise and flat-field response were characterized using a uniform light source constructed for this purpose. We show results in terms of spatial luminance noise and noise power spectrum for high-resolution images and for the same images processed with a pixel-aligned aperture. We find that the pixel-aligned aperture eliminates almost all the noise found in the high-resolution images, suggesting that most of the luminance noise in AMLCDs comes from the subpixel structure and less-than-100% aperture ratio, rather than from interpixel variations. PMID- 15124989 TI - Reconstructions in limited-view thermoacoustic tomography. AB - The limited-view problem is studied for thermoacoustic tomography, which is also referred to as photoacoustic or optoacoustic tomography depending on the type of radiation for the induction of acoustic waves. We define a "detection region," within which all points have sufficient detection views. It is explained analytically and shown numerically that the boundaries of any objects inside this region can be recovered stably. Otherwise some sharp details become blurred. One can identify in advance the parts of the boundaries that will be affected if the detection view is insufficient. If the detector scans along a circle in a two dimensional case, acquiring a sufficient view might require covering more than a pi-, or less than a pi-arc of the trajectory depending on the position of the object. Similar results hold in a three-dimensional case. In order to support our theoretical conclusions, three types of reconstruction methods are utilized: a filtered backprojection (FBP) approximate inversion, which is shown to work well for limited-view data, a local-tomography-type reconstruction that emphasizes sharp details (e.g., the boundaries of inclusions), and an iterative algebraic truncated conjugate gradient algorithm used in conjunction with FBP. Computations are conducted for both numerically simulated and experimental data. The reconstructions confirm our theoretical predictions. PMID- 15124990 TI - A treatment planning study comparing HDR and AGIMRT for cervical cancer. AB - The customization of brachytherapy dose distributions for gynecologic malignancies is limited by the spatial positioning of the applicators. We tested the hypothesis that applicator-guided intensity modulated radiation therapy (AGIMRT) has the potential to deliver highly conformal dose distributions to cervical tumors, representing improvement over distributions obtained with intracavitary brachytherapy. A commercial three-dimensional (3-D) treatment planning system was used to create plans for ten cervical cancer patients treated at our institution. Dose distributions of conventionally designed high dose rate (HDR) plans were compared against those of AGIMRT. Tumor delineation was based on a previously published binary threshold technique, using image intensity on positron emission tomography (PET) scans. AGIMRT treatment schedules were designed using two fraction sizes: 6.5 Gy, to directly reproduce the HDR fractionation, and 1.8 Gy, to simulate traditional external beam fractionation. The average minimum tumor dose was significantly greater for the AGIMRT dose distributions than for the HDR distributions (64.2 Gy vs 33.6 Gy; p = 0.005). The mean percent tumor volume at the prescription dose was higher for the AGIMRT plans (90.0% vs 58.2%; p = 0.005). Using AGIMRT, the mean percent volume at the tolerance limit was decreased for the bladder (6.1% vs 16.6%; p = 0.047) but increased for the rectum (4.1% vs 2.2%; p = 0.646). Our study suggests that there may be conceptual and dosimetric advantages to replacing HDR with AGIMRT for patients with large-volume cervical tumors. This investigation is being expanded using sequential PET images to model tumor regression and compare brachytherapy and AGIMRT throughout the course of therapy. PMID- 15124991 TI - Computerized characterization of breast masses on three-dimensional ultrasound volumes. AB - We are developing computer vision techniques for the characterization of breast masses as malignant or benign on radiologic examinations. In this study, we investigated the computerized characterization of breast masses on three dimensional (3-D) ultrasound (US) volumetric images. We developed 2-D and 3-D active contour models for automated segmentation of the mass volumes. The effect of the initialization method of the active contour on the robustness of the iterative segmentation method was studied by varying the contour used for its initialization. For a given segmentation, texture and morphological features were automatically extracted from the segmented masses and their margins. Stepwise discriminant analysis with the leave-one-out method was used to select effective features for the classification task and to combine these features into a malignancy score. The classification accuracy was evaluated using the area Az under the receiver operating characteristic (ROC) curve, as well as the partial area index Az(0.9), defined as the relative area under the ROC curve above a sensitivity threshold of 0.9. For the purpose of comparison with the computer classifier, four experienced breast radiologists provided malignancy ratings for the 3-D US masses. Our dataset consisted of 3-D US volumes of 102 biopsied masses (46 benign, 56 malignant). The classifiers based on 2-D and 3-D segmentation methods achieved test Az values of 0.87+/-0.03 and 0.92+/-0.03, respectively. The difference in the Az values of the two computer classifiers did not achieve statistical significance. The Az values of the four radiologists ranged between 0.84 and 0.92. The difference between the computer's Az value and that of any of the four radiologists did not achieve statistical significance either. However, the computer's Az(0.9) value was significantly higher than that of three of the four radiologists. Our results indicate that an automated and effective computer classifier can be designed for differentiating malignant and benign breast masses on 3-D US volumes. The accuracy of the classifier designed in this study was similar to that of experienced breast radiologists. PMID- 15124992 TI - AAPM TG-43 formalism for brachytherapy dose calculation of a 137Cs tube source. AB - We present a development of the use of the AAPM TG-43 dose formalism applied to 137Cs gynecological implant sources. The geometry factor, radial dose function, and anisotropy function of a 137Cs source modeled after the Nuclear Associates 67 809 series stainless steel jacketed tube source were derived following the AAPM TG-43 formalism. The dose rate distribution through the center of the source using the AAPM TG-43 dose formalism is calculated and compared with the calculations obtained using the Sievert summation and Monte Carlo simulation. The three methods resulted in an agreement within less than 5%, or an isodose rate line agreement within 2 mm. We demonstrate that the AAPM TG-43 formalism can be applied to 137Cs linear sources and is capable of serving as a 137Cs dose calculation algorithm that can be used for treatment planning purpose. PMID- 15124993 TI - Aperture maneuver with compelled breath (AMC) for moving tumors: a feasibility study with a moving phantom. AB - Respiration causes target motion, which is known to be one of the technical bottlenecks in radiotherapy, especially for stereotactic radio-surgery and intensity modulated radiotherapy (IMRT). To overcome this problem, aperture maneuver with compelled breath (AMC) has been developed. In order to simulate compelled respiratory motion, a moving phantom using a ventilator was designed. As the air flow was forced to the bellows, which simulates the lungs, by a ventilator, a film connected to the ventilator moved like the respiratory target motion. A software was developed to transfer multileaf collimator motion from breathless to actual periodic breathing conditions. Static fields as well as step and-shoot IMRT fields were modified in accordance with moving shapes to follow the target position, using the software with the controlled breathing information. Film dosimetry for a small field and for IMRT fields with a moving phantom was performed. To evaluate clinical implementation, five healthy volunteers were tested to breathe through a ventilator, and all of them could adapt the compelled breath without any difficulties. Additive margins for a moving target with AMC were not larger than 3 mm for respiratory organ motions up to 18 mm, while those with the static beam were 9 mm. For IMRT fields, large discrepancies were present between a static target and a moving target with the static beam, while they coincided well with AMC. Clinical acceptable differences between the dose distributions from a static target with the static beam and from a moving target with AMC revealed that this technique could be applied clinically. PMID- 15124994 TI - Influence of iodine contrast agent on the range of ion beams for radiotherapy. AB - The basis for the range calculation of heavy ions in tissue is an empirical correlation between x-ray CT numbers and ion ranges measured for tissue equivalent materials. Iodine contrast agents (CA), used during computed tomography (CT) imaging, lead to an increase of the Hounsfield units in tissue with increased CA uptake and cause errors in the calculation of the ranges. The aim of this work is to quantify how accurately ion range is calculated in CA loaded tissue. In order to quantify the mean change in Hounsfield units (HU), a statistical analysis of 25 CT data sets with and without CA was performed. To establish a relation between the change in Hounsfield units due to CA and changes in ion range, the Hounsfield number for various CA concentrations and the range of ions in CA was measured. The analysis of CT data sets showed that after intravenous injection of 100 ml contrast agent (Imeron300) at a concentration of 300 mg iodine/ml an increase of the Hounsfield numbers in tumor tissue of up to 57 HU can be observed. The measured range shift in CA is much smaller than calculated by the treatment planning system (TPS). The maximum error in range resulting from the CA enhanced data is approximately 2.5% and results mainly from this wrong interpretation of HU by the TPS. For a tumor with an extent of 5 cm this leads to an exaggeration of the ion ranges during irradiation of (1.24+/ 0.04) mm. This may be clinically relevant in cases where highest precision is needed and where organs at risk are close to the target volume. In view of these findings it may be safer to rely solely on native CT data for the purpose of dose and range optimization in therapy planning for heavy ions and protons. PMID- 15124995 TI - The significant effects of bone structure on inherent patient-specific DXA in vivo bone mineral density measurement inaccuracies. AB - An extended analytic exposition is developed of the effects bone structure has on the form and extent of systematic inaccuracies in planar dual-energy x-ray absorptiometry (DXA) in vivo bone mineral density (BMD) measurements. Explicit expressions for absolute and percentage BMD inaccuracies are derived and criteria governing these BMD inaccuracies delineated. It is shown that the effect of bone structure is to introduce a scale factor which modulates the sizable and unavoidable DXA in vivo/in situ BMD inaccuracies that arise directly from patient specific anthropometric and x-ray absorptiometric disparities among the several soft tissues present within the scan region of interest of any given bone site (i.e., lean muscle tissue, interposed and admixed fat, and red/yellow marrow combinations). Different magnitudes and patterns of BMD inaccuracies are shown to pertain for bone structures that are (i) essentially wholly trabecular, (ii) wholly cortical, and (iii) those containing both cortical and trabecular bone. Over the range of soft tissue anthropometrics typical of adult patients, the overall percentage inaccuracies in DXA-measured BMD are shown to be quite sizable and to vary considerably for different bone structures. For a typical lumbar vertebral bone site, BMD inaccuracies are found to be as large as approximately 25% for normal patients, to exceed approximately 35% for osteopenics, and to approach 50% for osteoporotic individuals. For bone sites with non-negligible cortical surrounds of trabecular structures (e.g., distal radius, some segments of proximal femur, etc.), it is shown that BMD percentage inaccuracies range up to approximately 20% for normal, approximately 25% osteopenic, and approximately 35% for osteoporotic patients. The BMD % inaccuracies associated with wholly cortical bone (trabecular-free) sites (e.g., mid-shaft femur, mid-shaft radius, etc.) are comparatively small, being less than approximately 2%. Depending on bone structure, bone size and shape, and patient-specific intra- and extra osseous soft tissue particulars of any given adult patient, DXA in vivo BMD measurements can be grossly inaccurate, and can severely under- or over-estimate the true value of BMD and mask or exaggerate true changes in BMD in ways not previously elucidated. It is concluded that in vivo DXA-measured and actual BMD cannot be considered to be synonymous, and clinical reliance upon the two being the same may readily conduce to seriously flawed and misleading diagnostic, prognostic, and prospective results. PMID- 15124996 TI - Computer-aided diagnosis scheme for histological classification of clustered microcalcifications on magnification mammograms. AB - The histological classification of clustered microcalcifications on mammograms can be difficult, and thus often require biopsy or follow-up. Our purpose in this study was to develop a computer-aided diagnosis scheme for identifying the histological classification of clustered microcalcifications on magnification mammograms in order to assist the radiologists' interpretation as a "second opinion." Our database consisted of 58 magnification mammograms, which included 35 malignant clustered microcalcifications (9 invasive carcinomas, 12 noninvasive carcinomas of the comedo type, and 14 noninvasive carcinomas of the noncomedo type) and 23 benign clustered microcalcifications (17 mastopathies and 6 fibroadenomas). The histological classifications of all clustered microcalcifications were proved by pathologic diagnosis. The clustered microcalcifications were first segmented by use of a novel filter bank and a thresholding technique. Five objective features on clustered microcalcifications were determined by taking into account subjective features that experienced the radiologists commonly use to identify possible histological classifications. The Bayes decision rule with five objective features was employed for distinguishing between five histological classifications. The classification accuracies for distinguishing between three malignant histological classifications were 77.8% (7/9) for invasive carcinoma, 75.0% (9/12) for noninvasive carcinoma of the comedo type, and 92.9% (13/14) for noninvasive carcinoma of the noncomedo type. The classification accuracies for distinguishing between two benign histological classifications were 94.1% (16/17) for mastopathy, and 100.0% (6/6) for fibroadenoma. This computerized method would be useful in assisting radiologists in their assessments of clustered microcalcifications. PMID- 15124998 TI - Accurate determination of dose-point-kernel functions close to the origin using Monte Carlo simulations. AB - Dose-point-kernel (DPK) functions are used extensively for the dosimetry of gamma and beta emitters in many physical problems. These functions are usually obtained from Monte Carlo simulations where the energy deposited in concentric spherical shells around a point source is tallied. The energy scored in a spherical shell divided by the shell mass is taken as the dose at some effective radius R(eff) of the shell. The effective radius R(eff), defined as the distance of a hypothetical zero-thickness scoring region from the source, can be evaluated in different ways for a finite thickness scoring region. For a shell thickness that is very small compared to the distance from the origin, this exact evaluation method becomes unimportant and the arithmetic mean is usually an accurate estimator for R(eff). However, accurately determining R(eff) can be problematic for the innermost regions when the radial dose function D(r) varies considerably over the finite spherical shell thickness. In this work, a new method for determining R(eff) is introduced which yields consistent results for any shell thickness, thus improving on previous Monte Carlo calculations for DPKs at or near the origin. Dimensionless DPK functions for monoenergetic electrons were reevaluated using EGSnrc with an emphasis on accuracy and consistency near the origin using our new method for determining R(eff). These improved functions were implemented in a software code to calculate the DPKs for an exhaustive list of 546 beta emitters, thus extending the compilation from previous works. PMID- 15124997 TI - Simulation study of a quasi-monochromatic beam for x-ray computed mammotomography. AB - The purpose of this simulation study was to evaluate the feasibility, benefits, and potential operating parameters of a quasi-monochromatic beam from a tungsten target x-ray source yielding projection images. The application is intended for newly developed cone beam computed mammotomography (CmT) of an uncompressed breast. The value of a near monochromatic x-ray source for a fully 3D CmT application is the expected improved ability to separate tissues with very small differences in attenuation coefficients. The quasi-monochromatic beam is expected to yield enhanced tomographic image quality along with a low dose, equal to or less than that of dual view x-ray mammography. X-ray spectra were generated with a validated projection x-ray simulation tool (XSpect) for a range of tungsten tube potentials (40-100 kVp), filter materials (Z=51-65), and filter thicknesses (10th to 1000th value layer determined at 60 kVp). The breast was modeled from ICRU-44 breast tissue specifications, and a breast lesion was modeled as a 0.5 cm thick mass. The detector was modeled as a digital flat-panel detector with a 0.06 cm thick CsI x-ray absorption layer. Computed figures of merit (FOMs) included the ratio of mean beam energy post-breast to pre-breast and the ratio of lesion contrasts for edge-located and center-located lesions as indices of breast beam hardening, and SNR2/exposure and SNR2/dose as indices of exposure and dose efficiencies. The impact of optimization of these FOMs on lesion contrast is also examined. For all simulated filter materials at each given attenuation thickness [10th, 100th, 500th, 1000th value layers (VLs)], the mean and standard deviation of the pre-breast spectral full-width at tenth-maximum (FWTM) were 16.1 +/- 2.4, 10.3 +/- 2.2, 7.3 +/- 1.4, and 6.5 +/- 1.5 keV, respectively. The change in beam width at the tenth maximum from pre-breast to post-breast spectra ranged from 4.7 to 1.1 keV, for the thinnest and thickest filters, respectively. The higher Z filters (Z=57-63) produced a quasi-monochromatic beam that allowed the widest tube potential operating range (50-70 kVp) while maintaining minimal beam hardening and maximal SNR2/exposure and SNR2/dose, and providing a contrast greater than that obtained in the unfiltered case. Figures of merit improved with increasing filter thickness, with diminishing returns beyond the 500th value layer attenuation level. Operating parameters required to produce optimal spectra, while keeping exposures equal to that of dual view mammography, are within the capability of the commercial x-ray tube proposed for our experimental study, indicating that use of these highly attenuating filters is viable. Additional simulations comparing Mo/Mo, Mo/Rh, and W/Rh target/filter combinations indicate that they exhibit significantly lower SNR2/exposure than the present approach, precluding them from being used for computed mammotomography, while maintaining dose limitations and obtaining sufficient SNR. Beam hardening was also much higher in the existing techniques (17%-42%) than for our technique (2%). Simulations demonstrate that this quasi-monochromatic x-ray technique may enhance tissue separation for a newly developed cone beam computed mammotomography application for an uncompressed breast. PMID- 15124999 TI - The stability of liquid-filled matrix ionization chamber electronic portal imaging devices for dosimetry purposes. AB - This study was performed to determine the stability of liquid-filled matrix ionization chamber (LiFi-type) electronic portal imaging devices (EPID) for dosimetric purposes. The short- and long-term stability of the response was investigated, as well as the importance of factors influencing the response (e.g., temperature fluctuations, radiation damage, and the performance of the electronic hardware). It was shown that testing the performance of the electronic hardware as well as the short-term stability of the imagers may reveal the cause of a poor long-term stability of the imager response. In addition, the short-term stability was measured to verify the validity of the fitted dose-response curve immediately after beam startup. The long-term stability of these imagers could be considerably improved by correcting for room temperature fluctuations and gradual changes in response due to radiation damage. As a result, the reproducibility was better than 1% (1 SD) over a period of two years. The results of this study were used to formulate recommendations for a quality control program for portal dosimetry. The effect of such a program was assessed by comparing the results of portal dosimetry and in vivo dosimetry using diodes during the treatment of 31 prostate patients. The improvement of the results for portal dosimetry was consistent with the deviations observed with the reproducibility tests in that particular period. After a correction for the variation in response of the imager, the average difference between the measured and prescribed dose during the treatment of prostate patients was -0.7%+/-1.5% (1 SD), and -0.6%+/-1.1% (1 SD) for EPID and diode in vivo dosimetry, respectively. It can be concluded that a high stability of the response can be achieved for this type of EPID by applying a rigorous quality control program. PMID- 15125000 TI - Introduction of a novel dose saving acquisition mode for the PortalVision aS500 EPID to facilitate on-line patient setup verification. AB - In external beam radiotherapy, electronic portal imaging becomes more and more an indispensable tool for the verification of the patient setup. For the safe clinical introduction of high dose conformal radiotherapy like intensity modulated radiation therapy, on-line patient setup verification is a prerequisite to ensure that the planned dosimetric coverage of the tumor volume is actually realized in the patient. Since the direction of setup fields often deviates from the direction of the treatment beams, extra dose is delivered to the patient during the acquisition of these portal images which may reach clinical relevance. The aim of this work was to develop a new acquisition mode for the PortalVision aS500 electronic portal imaging device from Varian Medical Systems that allows one to take portal images with reduced dose while keeping good image quality. The new acquisition mode, called RadMode, selectively enables and disables beam pulses during image acquisition allowing one to stop wasting valuable dose during the initial acquisition of "reset frames." Images of excellent quality can be taken with 1 MU only. This low dose per image facilitates daily setup verification with considerably reduced extra dose. PMID- 15125001 TI - Absolute dosimetry in a d(14 MeV) + Be fast neutron beam. AB - Since 1978, the Universitatsklinikum in Essen operates a d(14 MeV) + Be fast neutron beam for patient treatment. Dosimetric studies were performed in a rectangular 40 x 40 mm2 neutron/photon field using a transportable water calorimeter, which had been developed at the Physikalisch-Technische Bundesanstalt. The water calorimeter allowed small dosimeters to be directly calibrated in units of absorbed dose-to-water in a cylindrical phantom of 50 mm in diameter. Also, the twin detector method was applied in order to determine the photon and the neutron dose separately. By making use of a calibrated ionization chamber, the absorbed dose-to-water calibration in the cylindrical water phantom was transferred to a water phantom, a cube 300 mm on a side. Experiments and Monte Carlo calculations covering the neutron producing target, the collimator and the influence of the water calorimeter on the spectral neutron fluence at the measurement position allow the relative uncertainty of the absorbed dose-to-water determination to be reduced to 2.6% (1 SD). This direct absorbed dose-to-water determination by calorimetry has shown that the treatment planning system underestimates the physical dose to tissue by 9%. For clinical purposes, the statement of the prescribed dose had to be increased by 9% in order that the absolute absorbed dose remains constant and that the same biological endpoints are reached. PMID- 15125002 TI - Segmentation of nodules on chest computed tomography for growth assessment. AB - Several segmentation methods to evaluate growth of small isolated pulmonary nodules on chest computed tomography (CT) are presented. The segmentation methods are based on adaptively thresholding attenuation levels and use measures of nodule shape. The segmentation methods were first tested on a realistic chest phantom to evaluate their performance with respect to specific nodule characteristics. The segmentation methods were also tested on sequential CT scans of patients. The methods' estimation of nodule growth were compared to the volume change calculated by a chest radiologist. The best method segmented nodules on average 43% smaller or larger than the actual nodule when errors were computed across all nodule variations on the phantom. Some methods achieved smaller errors when examined with respect to certain nodule properties. In particular, on the phantom individual methods segmented solid nodules to within 23% of their actual size and nodules with 60.7 mm3 volumes to within 14%. On the clinical data, none of the methods examined showed a statistically significant difference in growth estimation from the radiologist. PMID- 15125003 TI - EUD-based margin selection in the presence of set-up uncertainties. AB - To assess the impact of geometric uncertainties on treatment plan design, we have performed a numerical simulation in which both systematic and random errors were included. A clinical target volume (CTV) with an abutting organ at risk (OAR), both of 50 mm diameter, in a cubic phantom was modeled. A four-field conformal treatment plan was designed in which one pair of parallel-opposed beams traversed the OAR and CTV while the other pair intersected the CTV only. Field size, prescribed (isocenter) dose and systematic set-up uncertainty were varied in two orthogonal directions to examine their impact on the outcome as predicted by the dose volume histogram (DVH) and the phenomenological form of equivalent uniform dose (EUD). Of the systematic uncertainty levels considered (0, 2, 4, and 6 mm standard deviations of a Gaussian distribution), 10 mm margin (CTV-PTV) was adequate to maintain the integrity of the dose distribution within the CTV. However, reducing the margin (and hence field size) without reducing set-up errors required an increase in the isocenter dose to compensate for the loss in EUD. It was found that, in the direction containing both the CTV and OAR, with random and systematic uncertainties of 2 and 4 mm respectively, increasing the isocenter dose by about 3.5 Gy on a 6 mm-margin plan resulted in the statistically equivalent EUD value to that with a 10 mm-margin for the CTV, while the OAR EUD is dropped by 1 Gy. In general, though, the directional sensitivity to geometric uncertainties, and hence the required margin size in different directions, was dependent on beam geometries and the relative positions of the structures under consideration relative to the beam directions. Based on the validity of the EUD concept, our general conclusion is that modest dose escalation may result in plans that better achieve clinical objectives. Also, a simple single number plan quality index such as EUD5%, discussed in the paper, facilitates meaningful statistical comparisons between competing treatment strategies. PMID- 15125004 TI - Computerized detection of colorectal masses in CT colonography based on fuzzy merging and wall-thickening analysis. AB - In recent years, several computer-aided detection (CAD) schemes have been developed for the detection of polyps in CT colonography (CTC). However, few studies have addressed the problem of computerized detection of colorectal masses in CTC. This is mostly because masses are considered to be well visualized by a radiologist because of their size and invasiveness. Nevertheless, the automated detection of masses would naturally complement the automated detection of polyps in CTC and would produce a more comprehensive computer aid to radiologists. Therefore, in this study, we identified some of the problems involved with the computerized detection of masses, and we developed a scheme for the computerized detection of masses that can be integrated into a CAD scheme for the detection of polyps. The performance of the mass detection scheme was evaluated by application to clinical CTC data sets. CTC was performed on 82 patients with helical CT scanners and reconstruction intervals of 1.0-5.0 mm in the supine and prone positions. Fourteen patients (17%) had a total of 14 masses of 30-50 mm, and sixteen patients (20%) had a total of 30 polyps 5-25 mm in diameter. Four patients had both polyps and masses. Fifty-six of the patients (68%) were normal. The CTC data were interpolated linearly to yield isotropic data sets, and the colon was extracted by use of a knowledge-guided segmentation technique. Two methods, fuzzy merging and wall-thickening analysis, were developed for the detection of masses. The fuzzy merging method detected masses with a significant intraluminal component by separating the initial CAD detections of locally cap like shapes within the colonic wall into mass candidates and polyp candidates. The wall-thickening analysis detected nonintraluminal masses by searching the colonic wall for abnormal thickening. The final regions of the mass candidates were extracted by use of a level set method based on a fast marching algorithm. False-positive (FP) detections were reduced by a quadratic discriminant classifier. The performance of the scheme was evaluated by use of a leave-one-out (round-robin) method with by-patient elimination. All but one of the 14 masses, which was partially cut off from the CTC data set in both supine and prone positions, were detected. The fuzzy merging method detected 11 of the masses, and the wall-thickening analysis detected 3 of the masses including all nonintraluminal masses. In combination, the two methods detected 13 of the 14 masses with 0.21 FPs per patient on average based on the leave-one-out evaluation. Most FPs were generated by extrinsic compression of the colonic wall that would be recognized easily and quickly by a radiologist. The mass detection methods did not affect the result of the polyp detection. The results indicate that the scheme is potentially useful in providing a high-performance CAD scheme for the detection of colorectal neoplasms in CTC. PMID- 15125005 TI - Investigation of physical image quality indices of a bone densitometry system. AB - Osteoporosis is a disease characterized by a loss of bone mass and a deterioration of bone structure. Bone mineral density (BMD) measures bone mass and is currently the method used to diagnose osteoporosis, while computerized radiographic texture analysis (RTA) is being investigated as a measure of bone structure. The GE/Lunar PIXI peripheral bone densitometer (PD) system, which uses dual-energy subtraction to measure BMD, also provides a digital image of the heel or forearm. The goal of our current research was to evaluate the physical imaging properties of the PIXI system (pixel size of 0.2 mm) compared to a Fuji computed radiography (CR) system (pixel size of 0.1 mm) to determine its suitability for texture analysis from image data. Contrast was measured using a series of uniform images covering the useful clinical exposure range. Spatial resolution was characterized by the presampling modulation transfer function (MTF) determined by an edge method. Noise power spectra (NPS) for different exposures were calculated using a two-dimensional Fourier analysis method. The expectation modulation transfer function was measured and combined with the NPS data to calculate the noise-equivalent number of quanta. The slope of the characteristic curve of the peripheral densitometer (PD) system was found to be position dependent across the image, although this dependence was substantially reduced by use of the system's clinical-settings corrections. An MTF value of 0.5 was found at 0.5 cycles/mm for the densitometry system compared to the same value at 1.6 cycles/mm for the CR system. Unlike the CR system, the NPS of the densitometry system was found not to be directionally dependent and did not drop off at higher spatial frequencies. PMID- 15125006 TI - Comparison of radiographic texture analysis from computed radiography and bone densitometry systems. AB - Osteoporosis is a disease that results in an increased risk of bone fracture due to a loss of bone mass and deterioration of bone structure. Bone mineral density (BMD) provides a measure of bone mass and is frequently measured by bone densitometry systems to diagnose osteoporosis. In addition, computerized radiographic texture analysis (RTA) is currently being investigated as a measure of bone structure and as an additional diagnostic predictor of osteoporosis. In this study, we assessed the ability of a peripheral bone densitometry (PD) system to yield images useful for RTA. The benefit of such a system is that it measures BMD by dual-energy x-ray absorptiometry and therefore provides high- and low energy digital radiographic images. The bone densitometry system investigated was the GE/Lunar PIXI, which provides 512 x 512 digital images of the heel or forearm (0.2 mm pixels). We compared texture features of heel images obtained with this PD system to those obtained on a Fuji computed radiography (CR) system (0.1 mm pixels). Fourier and fractal-based texture features of images from 24 subjects who had both CR and BMD exams were calculated, and correlation between the two systems was analyzed. Fourier-based texture features characterize the magnitude, frequency content, and orientation of the trabecular bone pattern. Good correlation was found between the two modalities for the first moment (FMP) with r=0.71 (p value<0.0001) and for minimum FMP with r=0.52 (p value=0.008). Root mean-square (RMS) did not correlate with r=0.31 (p value>0.05), while the standard deviation of the RMS did correlate with r=0.79 (p value<0.0001). Good correlation was also found between the two modalities for the fractal-based texture features with r=0.79 (p value<0.0001) for the global Minkowski dimension and r=0.63 (p value=0.0007) for the fractal dimension from a box counting method. The PD system therefore may have the potential for yielding heel images suitable for RTA. PMID- 15125008 TI - Improving binding potential analysis in [11C]raclopride PET studies using cluster analysis. AB - To calculate binding potentials (BP) in [11C]raclopride brain PET studies a reference tissue model is widely used. The aim of the present study was to improve the determination of time activity curves (TAC) of reference tissue regions using cluster analysis. In four patients with Huntington disease TACs of a cerebellar reference region were calculated either from manually placed circular ROIs within the cerebellum or by cluster analysis. BP estimates derived from cluster analysis are independent from inter- and intraobserver variations and show an improved reproducibility combined with a low variability compared to manually placed cerebellar ROIs. This is of high value in longitudinal studies. PMID- 15125007 TI - Segmentation of IMRT plans for radical lung radiotherapy delivery with the step and-shoot technique. AB - The purpose of this work was to determine a segmentation protocol for the treatment of localized non-small-cell lung cancer (NSCLC) with intensity modulated radiotherapy (IMRT) that is as effective as possible while practically simple and hence robust to known practical inaccuracies. This study focused on the stratification of continuous profiles into a discrete number of intensity levels. The selection of the segmentation parameters for the delivery of the fluence profiles using multiple static fields has been considered. Five-field equispaced IMRT treatment plans of five patients with NSCLC were selected. The study comprised nine treatment plans for each patient, starting from a conformal plan, optimizing it for IMRT and then segmenting it utilizing different numbers of segments in each case and optimizing for segment weights separately. A conformal plan, optimized for beam directions, collimator and wedge angles, was also used for comparison with the IMRT plans, so as to consider the best coplanar conformal case. A dose objective for the PTV and the organs-at-risk plus a constraint for the spinal cord were set for all inverse plans. All stages were compared with the aid of dose-volume histograms, dose distributions at the plane of the isocenter, intensity maps for key beams and plots of PTV homogeneity and overall conformality versus complexity. The unsegmented IMRT plans gave the best results but cannot be realized in practice with an MLC. They were best approximated by plans that needed 106-167 segments to deliver, but did not deteriorate significantly when approximated by plans which required 26-40 segments in total. All segmented IMRT plans gave a better lung sparing than the conformal plans, indicating that the deterioration of IMRT plans following segmentation is not equivalent to that of unmodulated, conformal plans. However, optimized conformal plans have the potential to approach the lung sparing achieved by segmented IMRT plans. Among the IMRT situations examined, five-field treatment plans for the lung, utilizing a maximum of 40 segments in total, have proven to give a good approximation of the IMRT plans with continuous modulation. PMID- 15125009 TI - Influence of initial electron beam parameters on Monte Carlo calculated absorbed dose distributions for radiotherapy photon beams. AB - Our aim in the present study was to investigate the effects of initial electron beam characteristics on Monte Carlo calculated absorbed dose distribution for a linac 6 MV photon beam. Moreover, the range of values of these parameters was derived, so that the resulted differences between measured and calculated doses were less than 1%. Mean energy, radial intensity distribution and energy spread of the initial electron beam, were studied. The method is based on absorbed dose comparisons of measured and calculated depth-dose and dose-profile curves. All comparisons were performed at 10.0 cm depth, in the umbral region for dose profile and for depths past maximum for depth-dose curves. Depth-dose and dose profile curves were considerably affected by the mean energy of electron beam, with dose profiles to be more sensitive on that parameter. The depth-dose curves were unaffected by the radial intensity of electron beam. In contrast, dose profile curves were affected by the radial intensity of initial electron beam for a large field size. No influence was observed in dose-profile or depth-dose curves with respect to energy spread variations of electron beam. Conclusively, simulating the radiation source of a photon beam, two of the examined parameters (mean energy and radial intensity) of the electron beam should be tuned accurately, so that the resulting absorbed doses are within acceptable precision. The suggested method of evaluating these crucial but often poorly specified parameters may be of value in the Monte Carlo simulation of linear accelerator photon beams. PMID- 15125010 TI - Dose rate and SDD dependence of commercially available diode detectors. AB - The dose-rate dependence of commercially available diode detectors was measured under both high instantaneous dose-rate (pulsed) and low dose rate (continuous, Co-60) radiation. The dose-rate dependence was measured in an acrylic miniphantom at a 5-cm depth in a 10 x 10 cm2 collimator setting, by varying source-to detector distance (SDD) between at least 80 and 200 cm. The ratio of a normalized diode reading to a normalized ion chamber reading (both at SDD=100 cm) was used to determine diode sensitivity ratio for pulsed and continuous radiation at different SDD. The inverse of the diode sensitivity ratio is defined as the SDD correction factor (SDD CF). The diode sensitivity ratio increased with increasing instantaneous dose rate (or decreasing SDD). The ratio of diode sensitivity, normalized to 4000 cGy/s, varied between 0.988 (1490 cGy/s)-1.023 (38,900 cGy/s) for unirradiated n-type Isorad Gold, 0.981 (1460 cGy/s)-1.026 (39,060 cGy/s) for unirradiated QED Red (n type), 0.972 (1490 cGy/s)-1.068 (38,900 cGy/s) for preirradiated Isorad Red (n type), 0.985 (1490 cGy/s)-1.012 (38,990 cGy/s) for n type Pt-doped Isorad-3 Gold, 0.995 (1450 cGy/s)-1.020 (21,870 cGy/s) for n-type Veridose Green, 0.978 (1450 cGy/s)-1.066 (21,870 cGy/s) for preirradiated Isorad p Red, 0.994 (1540 cGy/s)-1.028 (17,870 cGy/s) for p-type preirradiated QED, 0.998 (1450 cGy/s)-1.003 (21,870 cGy/s) for the p-type preirradiated Scanditronix EDP20(3G), and 0.998 (1490 cGy/s)-1.015 (38,880 cGy/s) for Scanditronix EDP10(3G) diodes. The p-type diodes do not always show less dose-rate dependence than the n type diodes. Preirradiation does not always reduce diode dose-rate dependence. A comparison between the SDD dependence measured at the surface of a full scatter phantom and that in a miniphantom was made. Using a direct adjustment of radiation pulse height, we concluded that the SDD dependence of diode sensitivity can be explained by the instantaneous dose-rate dependence if sufficient buildup is provided to eliminate electron contamination. An energy independent empirical formula was proposed to fit the dose-rate dependence of diode sensitivity. PMID- 15125011 TI - Accounting for center-of-mass target motion using convolution methods in Monte Carlo-based dose calculations of the lung. AB - We have applied convolution methods to account for some of the effects of respiratory induced motion in clinical treatment planning of the lung. The 3-D displacement of the GTV center-of-mass (COM) as determined from breath-hold exhale and inhale CT scans was used to approximate the breathing induced motion. The time-course of the GTV-COM was estimated using a probability distribution function (PDF) previously derived from diaphragmatic motion [Med. Phys. 26, 715 720 (1990)] but also used by others for treatment planning in the lung [Int. J. Radiat. Oncol., Biol., Phys. 53, 822-834 (2002); Med. Phys. 30, 1086-1095 (2003)]. We have implemented fluence and dose convolution methods within a Monte Carlo based dose calculation system with the intent of comparing these approaches for planning in the lung. All treatment plans in this study have been calculated with Monte Carlo using the breath-hold exhale CT data sets. An analysis of treatment plans for 3 patients showed substantial differences (hot and cold spots consistently greater than +/- 15%) between the motion convolved and static treatment plans. As fluence convolution accounts for the spatial variance of the dose distribution in the presence of tissue inhomogeneities, the doses were approximately 5% greater than those calculated with dose convolution in the vicinity of the lung. DVH differences between the static, fluence and dose convolved distributions for the CTV were relatively small, however, larger differences were observed for the PTV. An investigation of the effect of the breathing PDF asymmetry on the motion convolved dose distributions showed that reducing the asymmetry resulted in increased hot and cold spots in the motion convolved distributions relative to the static cases. In particular, changing from an asymmetric breathing function to one that is symmetric results in an increase in the hot/cold spots of +/- 15% relative to the static plan. This increase is not unexpected considering that the target spends relatively more time at inhale as the asymmetry decreases (note that the treatment plans were generated using the exhale CT scans). PMID- 15125012 TI - Correlation between mammographic density and volumetric fibroglandular tissue estimated on breast MR images. AB - Previous studies have found that mammographic breast density is highly correlated with breast cancer risk. Therefore, mammographic breast density may be considered as an important risk factor in studies of breast cancer treatments. In this paper, we evaluated the accuracy of using mammograms for estimating breast density by analyzing the correlation between the percent mammographic dense area and the percent glandular tissue volume as estimated from MR images. A dataset of 67 cases having MR images (coronal 3-D SPGR T1-weighted pre-contrast) and corresponding 4-view mammograms was used in this study. Mammographic breast density was estimated by an experienced radiologist and an automated image analysis tool, Mammography Density ESTimator (MDEST) developed previously in our laboratory. For the estimation of the percent volume of fibroglandular tissue in breast MR images, a semiautomatic method was developed to segment the fibroglandular tissue from each slice. The tissue volume was calculated by integration over all slices containing the breast. Interobserver variation was measured for 3 different readers. It was found that the correlation between every two of the three readers for segmentation of MR volumetric fibroglandular tissue was 0.99. The correlations between the percent volumetric fibroglandular tissue on MR images and the percent dense area of the CC and MLO views segmented by an experienced radiologist were both 0.91. The correlation between the percent volumetric fibroglandular tissue on MR images and the percent dense area of the CC and MLO views segmented by MDEST was 0.91 and 0.89, respectively. The root mean-square (rms) residual ranged from 5.4% to 6.3%. The mean bias ranged from 3% to 6%. The high correlation indicates that changes in mammographic density may be a useful indicator of changes in fibroglandular tissue volume in the breast. PMID- 15125013 TI - Analysis of the penumbra enlargement in lung versus the quality index of photon beams: a methodology to check the dose calculation algorithm. AB - It is well known that considerable underdosage can occur at the edges of a tumor inside the lung because of the degradation of penumbra due to lack of lateral electronic equilibrium. Although present even at smaller energies, this phenomenon is more pronounced for higher energies. Apart from Monte Carlo calculation, most of the existing Treatment Planning Systems (TPSs) cannot deal at all, or with acceptable accuracy, with this effect. A methodology has been developed for assessing the dose calculation algorithms in the lung region where lateral electronic disequilibrium exists, based on the Quality Index (QI) of the incident beam. A phantom, consisting of layers of polystyrene and lung material, has been irradiated using photon beams of 4, 6, 15, and 20 MV. The cross-plane profiles of each beam for 5x5, 10x10, and 25x10 fields have been measured at the middle of the phantom with the use of films. The penumbra (20%-80%) and fringe (50%-90%) enlargement was measured and the ratio of the widths for the lung to that of polystyrene was defined as the Correction Factor (CF). Monte Carlo calculations in the two phantoms have also been performed for energies of 6, 15, and 20 MV. Five commercial TPS's algorithms were tested for their ability to predict the penumbra and fringe enlargement. A linear relationship has been found between the QI of the beams and the CF of the penumbra and fringe enlargement for all the examined fields. Monte Carlo calculations agree very well (less than 1% difference) with the film measurements. The CF values range between 1.1 for 4 MV (QI 0.620) and 2.28 for 20 MV (QI 0.794). Three of the tested TPS's algorithms could not predict any enlargement at all for all energies and all fields and two of them could predict the penumbra enlargement to some extent. The proposed methodology can help any user or developer to check the accuracy of its algorithm for lung cases, based on a simple phantom geometry and the QI of the incident beam. This check is very important especially when higher energies are used, as the inaccuracies in existing algorithms can lead to an incorrect choice of energy for lung treatment and consequently to a failure in tumor control. PMID- 15125014 TI - Differential regulation of MMP-13 by chemical modified tetracyclines in osteoblasts. AB - Tetracyclines have been shown to regulate matrix metalloproteinase (MMP) expression in numerous cell types with various periodontal disease models. MMP 13, or collagenase-3, has been shown to be induced by a number of osteotropic cytokines and hormones in osteoblastic cells. In this study, we studied MMP-13 gene expression and regulation in osteoblasts by chemically modified tetracyclines (CMTs). Preliminary cytotoxicity studies indicated that 1-10 microg/ml of CMT-8 did not result in statistically significant cell death. Additional fluorescent microscopy experiments indicated that CMT-8 but not CMT-5 had a nuclear distribution within one hour of addition CMTs. Using primary rat calvarial osteoblastic cells obtained from 21-day old neonatal rats, we determined MMP-13 gene expression when stimulated by parathyroid hormone (PTH), interleukin (IL)-1b, or tumour necrosis factor (TNF)-alpha in the presence or absence of CMT-8 or -5. Semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) using total RNA was used to determine relative expression of MMP-13 compared to glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a constitutively expressed housekeeping gene. Results indicate that all agents were consistently able to induce MMP-13 expression in primary calvarial osteoblastic cells. However, CMT-8 inhibited MMP-13 expression from IL-1b and PTH stimulated cells while having little effect on TNF-alpha stimulated MMP-13 expression. In contrast, CMT-5 only inhibited PTH stimulated MMP-13 expression, with no effect on IL-1b or TNF-alpha stimulated MMP-13 expression. These results suggest that CMT-8 and CMT-5 may differentially affect cell signaling pathways in osteoblastic cells that mediate MMP-13 gene expression. PMID- 15125015 TI - Antibiotic prescription and dental practice within Saudi Arabia; the need to reinforce guidelines and implement specialty needs. AB - The aim of the present survey was to analyse the frequency and type of antibiotic prescriptions used in dental practice in Saudi Arabia, to assess the awareness of periodontal treatment need among dentists and specialists and to evaluate the distribution of dental specialties in Saudi Arabia. We distributed a questionnaire to 378 dentists including different specialists working in various major hospitals and dental departments. This same questionnaire was then redistributed, 3 years later, to 211 participants. Results from both surveys indicated that antibiotics were not always prescribed on a scientific basis. Cost was the most significant factor (p < 0.05). Interestingly, and although periodontal disease was included as one of the possible causes of infections, the penicillin group was shown to be the group of choice (p < 0.007) and was prescribed by 45% of all participants, metronidazole and tetracyclines followed and were the second most commonly prescribed antibiotics in both surveys (p < 0.05). It was further demonstrated that although the need for periodontics is present among patients and specialists, it is one of the least practised specialties (p < or = 0.008) along with oral surgery (p < or = 0.36). PMID- 15125016 TI - The influence of gingival health status on periodontal probing measurements. A clinical study in humans. AB - This study aimed to clarify the extent to which gingival health status alters probing measurements. One hundred sites from 26 patients undergoing periodontal treatment were selected as follows: 50 sites without inflammation and 50 sites with bleeding on probing even after basic therapy. All sites required periodontal surgery and were submitted to two probings with a constant-force probe (0.25 N). The pre-surgical probing (S1) measured the relative attachment level. The second probing (S2), during an open flap procedure, in the same location as S1, measured the distance between the occlusal surface of the stent and the bone crest. The difference between the measurements of the two probings (S2 - S1) performed at each site determined the distance between the bottom of the clinical pocket and the bone crest (S3). The median and the mean of the S1 measurements were 1.60 mm and 1.64 +/- 1.23 mm in the healthy sites and 1.50 mm and 1.63 +/- 1.03 mm in the bleeding sites, respectively. There was no statistically significant difference between healthy and bleeding sites (Mann-Whitney test, p = 0.625). It may be concluded that the penetration of the constant-force periodontal probe (0.25 N) was not influenced by bleeding on probing. PMID- 15125017 TI - The effects of manuka honey on plaque and gingivitis: a pilot study. AB - Research has shown that manuka honey has superior antimicrobial properties that can be used with success in the treatment of wound healing, peptic ulcers and bacterial gastro-enteritis. Studies have already shown that manuka honey with a high antibacterial activity is likely to be non-cariogenic. The current pilot study investigated whether or not manuka honey with an antibacterial activity rated UMF 15 could be used to reduce dental plaque and clinical levels of gingivitis. A chewable "honey leather" was produced for this trial. Thirty volunteers were randomly allocated to chew or suck either the manuka honey product, or sugarless chewing gum, for 10 minutes, three times a day, after each meal. Plaque and gingival bleeding scores were recorded before and after the 21 day trial period. Analysis of the results indicated that there were statistically highly significant reductions in the mean plaque scores (0.99 reduced to 0.65; p=0.001), and the percentage of bleeding sites (48% reduced to 17%; p=0.001), in the manuka honey group, with no significant changes in the control group. CONCLUSION: These results suggest that there may be a potential therapeutic role for manuka honey confectionery in the treatment of gingivitis and periodontal disease. PMID- 15125018 TI - New perspectives on tooth development and the dental stem cell niche. AB - Adult stem cells have the capacity to self-renew and differentiate along multiple lineages in addition to contributing to ongoing tissue maintenance and regeneration after injury. They reside in specific locations called stem cell niches. In biology of the tooth, the discovery of dental epithelial stem cells in continuously growing teeth has been a recent breakthrough. The niche for the adult stem cells of these teeth is formed at the region of the apical end in tooth development. The region possesses a commonly specialized histological structure for the maintenance of adult stem cells and the production of various progenitor cells producing dental tissues. The molecular signals regulating the maintenance and cell fate decision of adult stem cells, such as Notch1, Lunatic fringe, fibroblast growth factor (FGF)-10, are expressed in the epithelial structure and the surrounding mesenchyme. Based on histological and molecular biological studies, we propose a new concept that the eternal tooth buds producing various dental progeny are formed at the apical end in the development of continuously growing teeth, and coin a new term of "apical bud" for indicating this specialized epithelial structure. Furthermore, the relationship between signaling centers and the expression of FGF-10 mRNA as the determinant of morphogenesis is discussed with an emphasis on tooth and limb development, taking note that the expression pattern of FGF-10 is an important key for understanding the mechanisms for the diversity of cusp patterns and between continuous and limited growth. PMID- 15125019 TI - Hoechst 33342 is a useful cell tracer for a long-term investigation of articular cartilage repair. AB - The repair process of a full-thickness osteochondral defect was observed in a rat model using Hoechst 33342 as a cell tracer. The osteochondral defect was created at the medial femoral condyle of the right knee joints of twelve 11 week old male rats. Three weeks after the surgery, Hoechst 33342 was injected into the same knee joints. Calcein, a marker of the mineralization front, was then injected subcutaneousely twice at seven days and one day before harvesting of the tissue. At six, ten, and fourteen weeks and one year after the surgery, femoral condyles were obtained from the operated knee joints, fixed by alcohol, and embedded in polymethylmethacrylate. The sections were examined by fluorescent and then light microscopy. In the lateral femoral condyle cartilage, Hoechst 33342 labeling of chondrocyte nuclei was observed in all layers of the intact cartilage, and the dye never infiltrated beneath the subchondral bone plate. At 6 weeks after the surgery, Hoechst 33342-positive cells were observed not only in the regenerated fibrous cartilage, but also in the newly formed mineralized tissue in the medial femoral condyle. Interestingly, Hoechst 33342 labeling remained undiminished even one year after the intra-articular injection. The findings of the present study suggest that intra-articular injection of Hoechst 33342 is a useful tracer for long-term investigations of chondrocyte differentiation in vivo. PMID- 15125021 TI - Three-dimensional architecture of elastin and collagen fiber networks in the human and rat lung. AB - Collagen and elastin fibers are the major components of the lung connective tissue, but their spatial organization has not been well documented. We have demonstrated the three-dimensional architecture of collagen and elastin fiber networks in the human and rat lung using scanning electron microscopy. These networks in their original forms were extracted by an alkali-water maceration technique and a formic acid treatment, respectively. The collagen fibers formed a continuum extending throughout the lung and pleura. They were condensed in the alveolar mouth and subdivided into smaller fibers in the alveolar septa, thus forming basket-like networks. Sizes of the alveolar pores in the collagen fiber network of the alveolar septa became larger with age. In the collapsed lung, collagen fibers in the alveolar mouths and septa took on wavelike configurations, while in the inflated lung they became straight. The elastin fibers also formed a continuum, rich in the alveolar mouths and poor in the alveolar septa, were quite straight without any wavelike configuration. Transmission electron microscopy showed that collagen and elastin fibers were intermingled, suggesting that both fiber systems may act as parallel mechanical elements to stress or strain applied. Our results suggest that at low levels of strain the wavy collagen fibers are easily extended to allow alveolar mouths and alveoli to expand, with most of the stress being borne by adjacent elastin fibers, while at higher levels collagen fibers become straight and limit any further distension of alveolar ducts and alveoli. The elastin fiber continuum appears to permit the lung to effectively recoil or retract. The present study has also shown that alveolar pores enlarge with age, suggesting that collagen remodeling may be related to the pathogenesis of emphysema. PMID- 15125020 TI - Induction of adipogenesis by the intrasplenic transplantation of chick serum clots. AB - Chick serum contains a factor that stimulates adipogenesis in Meckel's chondrocytes in vitro. The present study examined whether chick serum has a capacity for adipogenic induction in vivo, by transplanting serum clots (created by drying chick serum for up to 4 weeks) into mouse spleens. Specimens were harvested for histological analyses, which included light and electron microscopy and immunohistochemistry. The transplanted serum clots induced the appearance of lipid droplet-containing cells in splenic cords and sinus. Almost all the lipid droplet-containing cells were positive for sudan staining and consisted of multilocular lipid vacuoles. Immunostaining showed that the adipocytes induced by transplantation of the serum clots initially appeared as peroxisome proliferator activated receptor-gamma (PPARgamma)-positive cells and developed into leptin and alpha-glycerophosphate dehydrogenase (GPDH)-producing cells, in addition to type III collagen synthesis. Furthermore, double immunofluorescence staining revealed that the immunoreactivity for GPDH was detected not only in stromal cells but also in macrophages. It was thus confirmed that stromal cells and macrophages in the spleen contain lipid droplets as seen in intact white adipose cells. The present results suggest that chick serum contains factors for adipocyte induction not only in vitro but also in vivo, and that the adipogenic potential does not depend on the supplements used during the cell culture. PMID- 15125022 TI - Nerve plexuses in the trachea and extrapulmonary bronchi of the rat. AB - Intrinsic nerve plexuses of the rat trachea and extrapulmonary bronchi were examined by immunohistochemistry. Three nerve plexuses--peritracheal and peribronchial, intramuscular, and submucosal--were found in the wall of the trachea and bronchi. Nerve cell bodies were located in the peritracheal and peribronchial nerve plexuses. They occurred singly or formed ganglia in the plexus, and regional differences in cell numbers were found in the cervical and thoracic portions of the trachea and in the extrapulmonary bronchia. In total, 83.5 +/- 28.3 ganglia (mean +/- SD, 57-131, n=5) and 749.8 +/- 221.1 nerve cell bodies (540-1,080, n=5) were found in the nerve plexus. The mean densities of ganglia were 0.31, 0.97 and 1.15/mm2, and the mean densities of the nerve cell bodies were 1.82, 9.26 and 11.54/mm2 in the cervical region, thoracic region of trachea, and extrapulmonary bronchi, respectively. Almost all nerve cell bodies in ganglia were positive for choline acetyltransferase and neuropeptide Y (NPY), and a few cells were positive for vasoactive intestinal peptide (VIP). In addition, in cholinergic nerves, a few nerve fibers in the smooth muscles were positive for substance P (SP), calcitonin gene-related peptide (CGRP), and VIP, and a moderate number of fibers were positive for NPY. Tyrosine hydroxylase immunoreactive nerve fibers were observed around blood vessels and within nerve bundles in the tunica adventitia. In the epithelium, nerve fibers were positive for SP and CGRP. Our results indicate that postganglionic neurons form three layers of cholinergic plexuses in the rat trachea and extrapulmonary bronchi, and that all of these possess intrinsic and extrinsic peptidergic innervation. PMID- 15125023 TI - Immunocytochemical localization of secretogranin III in the endocrine pancreas of male rats. AB - Secretogranin III (SgIII), a member of the granin protein family, is expressed specifically in neuronal and endocrine cells. To examine the precise localization of SgIII in the endocrine pancreas, pancreatic tissues of rats were analyzed immunocytochemically with a polyclonal anti-serum raised against rat SgIII. By light microscopy of semithin sections, the immunoreactivity for SgIII was readily detected in pancreatic A- and B-cells, faintly so in D-cells, and not at all in the exocrine pancreas. By immunoelectron microscopy, immunogold particles indicative of SgIII were observed in the peripheral regions of secretory granules, and universally in the pancreatic endocrine cells. Morphometrical analyses indicated that SgIII is most preferentially localized in the periphery of the secretory granule among granins. These findings suggest that SgIII is closely associated with the secretory granule membrane, serving to anchor the aggregates of other soluble constituents to the membrane. PMID- 15125024 TI - The induction of enamel and dentin complexes by subcutaneous implantation of reconstructed human and murine tooth germ elements. AB - Tooth induction by xenogenic graft of reconstructed human tooth germ components has never been attempted. Here we report our first attempt at a transplantation of human tooth germ components, heterologously recombined with mouse dental epithelia, into immunocompromised animals. Human third molar tooth germs enucleated from young patients as prophylactic treatment for orthodontic reasons were collected. The whole or minced human dental papilla was reconstructed with human- or mouse molar enamel epithelium, and transplanted in the dorsal aspect of C.B-17/Icr-scid Jcl mice. The transplant of human dental papilla reconstructed with human enamel epithelium formed thin dentin and immature enamel layers by 3 to 4 weeks, but remained extremely small in quantity due to a shortage of epithelial components in the graft. The addition of E16 mouse molar enamel organs (n=10-12) to each graft augmented the formation of tooth germ-like structures, but the differentiation of mouse molar ameloblasts was suppressed. However, once a solid layer of mineralized dentin was established, mouse ameloblasts accelerated their differentiation, and completed the enamel matrix formation and maturation within the following 4 weeks, whereas human ameloblasts, which had interacted with human dental papilla, remained in the stage of matrix formation during the same period. These data imply that, in reconstructed transplants, the differentiation of mouse dental epithelia is restrained by putative suppressive factors derived from human dental papilla until they are separated by mineralized dentin layers that serve as a diffusion barrier. The mouse enamel organ nevertheless retains its own phenotypic characteristics and intrinsic timing of cell differentiation and function. PMID- 15125025 TI - Cytological and immunocytochemical characterization of the insulin secreting insulinoma cell line RINm5F. AB - The rat insulinoma cell line RINm5F, an insulin secreting pancreatic beta cell line, has been used as an attractive model for basic studies of the mechanisms of insulin secretion and, more recently, as a model for the development of alternative methods for the treatment of diabetes. To elucidate the cytological properties and expression patterns of hormones of the gastro-entero-pancreatic system, suspensions of RINm5F cells were investigated by various methods including immunocytochemistry on serial semithin sections, quantitative immunocytochemistry, routine electron microscopy, immuno-electron microscopy, in situ hybridization, and TUNEL technique. At the ultrastructural level, several phenotypes of RIm5F cells were characterized by differences in the number, shape, size, and density of their secretory granules. The most common type contained a mixture of round granules varying in size and electron density. A second type predominantly contained relatively large, moderately dense granules. Moreover, a minority of cells was characterized by the occurrence of polymorphous electron dense granules or the complete absence of any secretory granules. The immunohistochemical data showed that, among the established islet hormones, insulin was present in more than 50% of cells, whereas glucagon and somatostatin occurred only sporadically. Though cells positive for pancreatic polypeptide (PP) were not found, PP-related peptides (NPY and PYY) however could be detected in a minority of cells. The great majority of RINm5F cells were immunoreactive for chromogranin B (CgB), followed by insulin, chromogranin A (CgA), and serotonin (5 HT). In addition to intercellular differences in the density of immunostaining, numerous colocalizations of immunoreactivities were found, suggesting that RINm5F cells represent a mixture of subtypes concerning the individual pattern of hormone expression. The present results reveal a wide range of heterogeneity with respect to the morphology and especially the hormone content between individual RINm5F cells. PMID- 15125026 TI - Response to ATP is accompanied by a Ca2+ influx via P2X purinoceptors in the coronary arterioles of golden hamsters. AB - In the vascular wall, adenosine-5'-triphosphate (ATP) released along with noradrenaline from sympathetic nerve terminals is considered to play an important role in controlling intracellular calcium ion ([Ca2+]i) levels in arteries. The present study examined how vascular smooth muscle cells in coronary arterioles respond to ATP in relation to [Ca2+]i dynamics. For this purpose, the dynamics of [Ca2+]i in the coronary arterioles of golden hamsters was examined by real-time laser scanning confocal microscopy. This technique enabled the visualization of [Ca2+]i changes in response to ATP in the intact coronary arterioles, the ultrastructure of which was well preserved. It was shown that an increase in [Ca2+]i in the arteriole smooth muscle cells was elicited by ATP. While P1 purinoceptor agonists have no effect on this process, P2 purinoceptor agonists were found to induce a [Ca2+]i increase in the smooth muscle cells. Suramin (an antagonist of P2X and P2Y receptors) completely inhibited ATP-induced [Ca2+]i dynamics, but reactive blue 2 (a P2Y receptor antagonist) did not. Uridine-5' triphosphate (a P2Y receptor agonist) had no effect on [Ca2+]i, but alpha,beta methylene ATP (a P2X receptor agonist) caused a strong increase in [Ca2+]i. We conclude that smooth muscle cells of the hamster coronary arterioles possess P2X, but not P1 or P2Y purinoceptors. The smooth muscle cells probably respond to extracellular ATP via P2X purinoceptors, resulting in the contraction of the coronary arterioles. PMID- 15125027 TI - Neutrophil extravasation in rat mesenteric venules induced by the chemotactic peptide N-formyl-methionyl-luecylphenylalanine (fMLP), with special attention to a barrier function of the vascular basal lamina for neutrophil migration. AB - The present study was performed to investigate morphologically the process of neutrophil extravasation induced by the synthetic bacterial peptide N formylmethionyl-leucyl-phenylalanine (fMLP) in venules of the rat mesentery by the combined use of intravital microscopy and transmission electron microscopy (TEM). Special attention was given to the interaction of the neutrophils with the endothelial cells and endothelial basal lamina. By intravital microscopy, the rolling and adhesion of leukocytes were observed within 3 min in preparations activated by fMLP. Neutrophils remained in the vascular wall for more than 30 min even after penetration of the endothelium. In this period, neutrophils migrating between endothelial cells and their basal lamina were often observed by TEM. After 40 min application of fMLP, some of the migrating neutrophils parted from the vessel wall into the surrounding connective tissues. There were neutrophils which passed through the small pore of the basal lamina with a cytoplasmic constriction. These findings indicate that the endothelial basal lamina acts as a physical barrier against neutrophil extravasation, thus resulting in the transient retainment of neutrophils beneath the endothelial cells of the venular wall. PMID- 15125028 TI - Molecular and clinical studies of Dent's disease in Japan: biochemical examination and renal ultrasonography do not predict carrier state. AB - BACKGROUND: Dent's disease is an X-linked renal tubular disorder characterized by low-molecular-weight-proteinuria, hypercalciuria, nephrolithiasis and renal failure. The disease is due to inactivation of a renal chloride channel gene, CLCN5. We have investigated 3 unrelated Japanese families for CLCN5 mutations and assessed the carrier mothers biochemically and ultrasonogaraphically to ascertain whether these clinical examinations can predict the carrier state of the disease. MATERIAL AND METHODS: Twelve members from these families were studied biochemically and ultrasonographically. Leukocyte DNA from probands was used with CLCN5-specific primers for PCR amplification of the coding region and exon-intron boundaries, and the DNA sequences of the products determined to identify abnormalities in the gene. RESULTS: Three novel CLCN5 mutations consisting of a single base "A" insertion between nucleotides 590 and 591, a nonsense mutation (R28X) and a missense mutation (G506R) were exhibited. Hypophosphatemia was detected in 2 patients, beta2-microglobulinuria, alpha1-microglobulinuria, and hyperretinol binding proteinuria in 6 patients, hypercalciuria in 5 patients, decreased urine osmolality in 3 patients, and nephrocalcinosis or nephrolithiasis in 4 patients. Biochemical analysis of the urine and the renal ultrasonography in each carrier mother were completely normal. CONCLUSIONS: Neither urinary low molecular-weight-proteins, urinary calcium to creatinine ratio, nor renal ultrasonography was predictive of carrier state in the 3 families with this disease, although each carrier mother had CLCN5 mutation. Hypophosphatemia and decreased urine osmolality might be a hint to suspect the carrier state of Dent's disease, although these findings are not found frequently. PMID- 15125029 TI - Cardiovascular morbidity and mortality in patients with diabetes mellitus type I after kidney transplantation: a case-control study. AB - BACKGROUND: The proportion of diabetics among patients requiring renal replacement therapy continues to increase in most western countries. The acceptance rate for renal transplantation varies among transplant centers and is influenced by the current opinion on the outcome of transplantation in diabetics. Controlled data on patient and graft survival in type I diabetics, however, are scarce. METHODS: We performed a retrospective case-control analysis on patient and graft survival and the cardiovascular morbidity of patients with type I diabetes after renal transplantation versus carefully matched non-diabetic transplant recipients. Match criteria were duration of previous hemodialysis, age and date of renal transplantation. Moreover, risk factors for cardiovascular disease in uremic patients were evaluated at the time of registration for renal transplantation and at the end of the observation period. RESULTS: Seventy-seven matched pairs were enclosed. Patient survival was significantly worse in the diabetic patients, graft survival was comparable in both groups, when graft loss because of patient's death was censored. In the diabetic patients, risk of death (odds ratio: 4.38) as well as the prevalence of cardiovascular morbidity (odds ratio: 4.47) were significantly higher than in the matched nondiabetic controls. Cox regression analysis showed that diabetes mellitus was an independent risk factor for patient survival; no association was found with hypertension, hyperlipidemia, hyperparathyroidism, calcium x phosphate product, body mass index and HbA1c. Cardiovascular morbidity, however, was already significantly higher in the diabetic group at the time of registration. CONCLUSIONS: Diabetes mellitus type I has a dominant impact on morbidity and mortality after renal transplantation and is associated with an approximately 4-fold higher risk of death. Cardiovascular disease accounts for the significantly worse long-term outcome of diabetic patients after renal transplantation. PMID- 15125030 TI - Preemptive treatment with oral ganciclovir for pediatric renal transplantation. AB - This prospective study examines 42 children in the first year after renal transplantation. They all received intravenous ganciclovir prophylaxis for cytomegalovirus in the immediate post-transplant period. Quantitative antigenemia (pp68) determinations and blood, urine and throat cultures were done on a scheduled basis to detect cytomegalovirus. Infection was detected in 22 children (52.4%) within an average 44.31 +/- 27.38 days; 5/22 were symptomatic. The antigenemia was positive (+) in all the infected patients, and so were blood culture in 68.2%, urine culture in 59.1% and throat culture in 31.8%. A positive antigenemia was the earliest finding in all cases but 1. The 5 children with clinical symptoms received intravenous ganciclovir. Asymptomatic infected children received oral ganciclovir at an average dose of 47.64 +/- 8.10 mg/kg/day (median 46.58 (range 33-58.7) mg/kg/day) for an average of 58.47 +/- 27.76 days (median 58 (range 26-211) days). No patient developed disease or ganciclovir resistance during the treatment. No patient presented acute graft rejection or renal dysfunction and their glomerular filtrate rate at 1 year was similar to that of noninfected children (90.38 +/- 26.51 vs. 93.93 +/- 36.24 ml/min/1.73 m2). We conclude that preemptive treatment with oral ganciclovir is useful and safe in children with renal transplantation and that monitoring blood antigenemia is a sensitive and early method to detect and control CMV infection. PMID- 15125031 TI - PTH secretion in patients with chronic renal failure assessed by a modified CiCa clamp method: effects of 1-year calcitriol therapy. AB - BACKGROUND: Secondary hyperparathyroidism (secondary HPT) in patients with chronic renal failure (CRF) is characterized by parathyroid gland hyperplasia and an intrinsic defect in the recognition of parathyroid hormone (PTH) secretion. Conflicting results have been reported regarding the set point for calcium regulated PTH release and its modification by calcitriol therapy in hemodialysis patients. Additionally, the effect of calcitriol on the calcium/PTH relationship in predialysis CRF patients with early secondary HPT has not been investigated. Our objective in this controlled study was to investigate the calcium/PTH relationship and to determine the calcium set point in patients with early stages of CRF before and after a 1-year treatment with calcitriol and in normal volunteers. METHODS: Nine patients with an early stage of CRF (GFR between 20 and 50 ml/min x 1.73 m2 b.s.) aged 35-77 years and 13 healthy volunteers (HV) aged 26 60, years were included in the study. All participants were investigated by sequential lowering and raising of serum calcium levels comprising the following phases: blood-ionized calcium (Ca2+) was lowered by about 0.2 mmol/l (3 steps), steady-state hypocalcemia of Ca2+ 0.2 mmol/l below the baseline (step 4), stop of the infusion for 5 minutes (step 5), Ca2+ was raised to about 0.2 mmol/l above baseline (steps 6 and 7), and a steady state hypercalcemia of Ca2+ 0.2 mmol/l above baseline (step 8). Ionized calcium and intact PTH (iPTH) were measured at 30 time points during 240 minutes. The calcium set point was determined using the classical 4-parameter model. The CiCa clamp test was performed before and after a 1-year treatment with 0.5 microg of calcitriol thrice weekly. RESULTS: No differences in the set point were observed between HV and CRF patients with early secondary HPT. Four of 9 patients responded to calcitriol treatment with a decrease in basal serum iPTH levels ("responders"). There was no difference between renal function (GFR 18 +/- 6 vs. 17 +/- 8 ml/min x 1.73 m2 b.s.), set point (Ca2+ 1.07 +/- 0.13 vs. 1.07 +/- 0.06 mmol/l) and suppressibility of PTH secretion (PTHmin% 7.3 +/- 1.6 vs. 8.2 +/- 2.9) in responders vs non-responders, nor did these values change after treatment with calcitriol. PTHmin% decreased significantly in the whole group after treatment (10.4 +/- 8.5 vs. 7.8 +/- 2.4). CONCLUSIONS: Although the calcium set point was not different in predialysis CRF patients with early secondary HPT compared to HV, calcitriol treatment improved the calcium-related suppression of PTH secretion (PTHmin%). PMID- 15125032 TI - The effect of an exercise program during hemodialysis on dialysis efficacy, blood pressure and quality of life in end-stage renal disease (ESRD) patients. AB - AIM: We wished to determine if an 8-week program of exercise during dialysis in end-stage renal disease (ESRD) patients would increase urea removal (enhance dialysis efficacy) with subsequent improvements in work performance and perception of quality of life, and/or alterations in cardiovascular status. METHODS: Self-care hemodialysis patients (EX, n = 6) performed cycle ergometry exercise 3 times per week during their dialysis session at 40-50% maximal work capacity for 15 min during each of the first 3 hours of dialysis and were matched for age, protein catabolism rate, and WLmax with a CON group (n = 7). Dialysis efficacy was measured using serum urea clearance (Kt/V) and dialysate urea clearance (DUC) during the first 2 hours of dialysis. Resting blood pressure was monitored on a sessional basis, pre- and postdialysis and during exercise in the EX group. QOL, measured using the SF-36 questionnaire, and WLmax were determined prior to and at 4 and 8 weeks of the exercise program. RESULTS: DUC was significantly elevated in the EX group at the end of the exercise program, but was of insufficient magnitude to result in an overall increase in Kt/V. DUC decreased in the CON group but Kt/V remained unchanged. No changes in resting blood pressure occurred in either group over the course of the study, however, pulse pressure tended to increase in the CON group but decrease in the EX group, indicating a potential beneficial adaptation of the cardiovascular system in patients undergoing an exercise program. The exercise program had no effect on QOL scores and this was most likely due to the short duration of the exercise program and high-functioning level of the population studied as compared to normative data for this patient population. We also found that 33% of the exercise sessions in the 3rd hour of dialysis were not performed due to hypotensive events. CONCLUSION: Exercise during dialysis enhanced dialysate urea removal but not serum urea clearance. Alterations in the modality and the timing of exercise during dialysis may be required to elicit increases in serum urea clearance. It is also recommended that exercise during dialysis be performed during the first 2 hours of dialysis. PMID- 15125033 TI - Diabetes delaying the diagnosis of RPGN. AB - The present case describes the unusual association of a crescentic glomerulonephritis (GN), diabetes mellitus and a monoclonal gammopathy. After an unexplained deterioration of renal function, a kidney biopsy was performed. The finding of crescentic glomerulonephritis was unexpected. This case illustrates the usefulness of kidney biopsy in diabetes to exclude concomittant disease. PMID- 15125034 TI - Postinfectious diffuse proliferative glomerulonephritis and acute renal failure in an HIV patient. AB - Postinfectious proliferative glomerulonephritis may occur in HIV-infected patients, although it is not a common cause of severe acute renal failure in them. We report a woman with HIV infection, who developed hypocomplementemic acute nephritic syndrome 10 days after an upper respiratory infection. Systemic diseases were excluded. The serum creatinine level increased to 6.6 mg/dl. Renal biopsy showed diffuse endocapillary proliferative glomerulonephritis, with mesangial and capillary walls, granular deposits of IgG and C3 by immunofluorescence. She was given corticosteroids with progressive normalization of her renal function. No opportunistic infections have occurred during 1-year follow-up. PMID- 15125035 TI - An atypical course of Caroli's disease in a renal transplant patient--case report and review of the literature. AB - This is a rare case of Caroli's disease, diagnosed following renal transplantation in a patient with autosomal recessive polycystic kidneys. Despite advanced cystic transformation of the biliary tree with striking architectural changes, there was no evidence of portal hypertension or hepatic fibrosis. Moreover, the patient did not suffer a single episode of cholangitis, a most interesting feature of this case. Her clinical course was punctuated by repeated episodes of gastrointestinal and urinary tract infections with resistant organisms; but fortunately, she had no evidence of septicemia. Recurrent Salmonella gastroenteritis indicated a chronic carrier state with the dilated bile ducts possibly acting as a potential reservoir. This has significant implications considering the immune suppression associated with renal transplantation. In general, Caroli's disease is rare. Therefore, a high index of suspicion for the diagnosis of Caroli's disease is warranted especially in patients with ARPKD or ADPKD. Once confirmed, affected patients with end-stage renal disease such as our patient, should ideally undergo combined liver-kidney transplantation. PMID- 15125036 TI - Brown tumors developing in renal transplant recipients with persistent hyperparathyroidism: two case reports and review of literature. AB - BACKGROUND: Brown tumors, evidence for severe hyperparathyroidism, are rare in end-stage renal disease, and are distinctly uncommon in patients with a functioning renal transplant. CASE REPORTS: We report two cases of brown tumors developing after renal transplantation, and discuss their presentation and treatment. We review the literature. CONCLUSIONS: We suggest that persistent hyperparathyroidism post-renal transplant requires aggressive intervention to avoid significant consequences. PMID- 15125037 TI - Membranous nephropathy accompanied by HHV8-DNA-positive angiolymphoid hyperplasia of the skin with eosinophilia: lack of HHV8 viral DNA in the kidney biopsy. PMID- 15125038 TI - Spontaneous remission in a case of severe crescentic IgA nephropathy without long term treatment. PMID- 15125039 TI - Morphology, development and comparative anatomical evaluation of the testicular excretory pathway in Acipenser. AB - In the differentiated state, the testicular excurrent duct system of the sturgeon begins as a longitudinal marginal network of the testis, extending along the entire length of the male gonad. From here, mesorchial transversal ducts travel to the ventral aspect of the pars sexualis of the opisthonephros where they merge behind the dorsal coelomic wall to form the longitudinal marginal network of the kidney. Then, the seminal pathways enter the confines of the pars sexualis of the opisthonephros and divide into a complicated, multipartite system consisting of (1) centropapillary ducts, situated in the center of a group of urinary collecting ducts, (2) lacunary basal sinuses, located on the bases of opisthonephric columns and (3) intracolumnar ducts running inside the renal columns, the latter representing typical functional units of the adult sturgeon kidney. The contacts between intracolumnar ducts and the vascular poles of corresponding renal corpuscles represent the urogenital junction in the sturgeon. The nephrons of the pars sexualis involved in sperm transport do not lose their urinary functions, but are histologically identical to those of the pars excretoria which are solely urinary. The opisthonephros of sturgeons grows continuously by the formation of new nephrons from an opisthonephric blastema located on the base of each renal column. A close topographical association between this blastema tissue and the lacunary basal sinuses of the testicular excurrent duct system guarantees that new renal corpuscles in the pars sexualis are included in the seminal passage from their beginning. From the urogenital junctions, on their way to the exterior, the spermatozoa have to travel through Bowman's capsules and tubules of the nephrons involved, then through the urinary collecting ducts, the wolffian duct and finally the sinus urogenitalis. The development of the testicular excurrent duct system begins in 8-month-old animals in the pregonadal area of the gonadal fold. Here, a primary gonoductal blastema proliferates to form a longitudinal network of anastomosing strands, situated in the dorsal mesogonadal attachment. From this primary longitudinal network, small tubules grow into the direction of the opisthonephros and into the direction of the testis. In the period from 8 to 18 months, the testicular excurent duct system reaches the adult state. In conclusion, the testicular excurrent ducts of sturgeons initially develop similar to those of Polypterus and in modern teleosts from a primary longitudinal system, beginning in the pregonadal area, localized in the mesogonadal attachment and extending caudally. Then, in a second step of development, the phylogenetically older situation, using parts of the kidney as passage, already seen in Chondrichthyes, but preserved also in higher vertebrates, is achieved in Acipenser. For this, seminal ducts grow into the opisthonephros and establish here the urogenital junctions with corresponding renal corpuscles. Furthermore, the initially longitudinally oriented ducts in the mesogonadal attachment partly lose their continuity and become integrated into the course of the transversal mesorchial ducts, represented by their portions with the widest lumina and the thickest walls. PMID- 15125040 TI - Electron microscopic alterations of the rat's pleura after experimental haemothorax. AB - The initial signs of pleural reactivity and the subsequent mechanisms of pleural healing still remain unsolved. The visceral and parietal (costal and diaphragmatic) pleura were investigated following an experimental haemothorax (EH) by transmission electron microscopy. Young-adult Wistar rats were divided in five groups and survived 6 hours, 1, 3, 8 and 15 days respectively after EH. Six hours after EH the mesothelial cells had a more prominent lysosomal system and electron-dense material in the vesicles, as in the dilatated intercellular spaces. On the 1st day of the EH the mesothelial cytoplasm formed a thin interrupted band. The extravasal cells built multiple layers over the basal lamina, leading to a thicker submesothelial layer, occupying the superficial position toward the pleural cavity. The activated mesothelial cells covered both pleural sheets on the 3rd day after EH. Eight days after EH different membrane bodies, large apical evaginations, elastic-like formations, an extensive vesicular and cytofilamentous systems characterized the mesothelium. The wider elastic membrane showed thickenings, protrusions, bifurcations and double course. Fifteen days after EH larger zones in both pleural sheets displayed thinner basal lamina, remnants of elastic membrane and a thicker submesothelial layer. In conclusion, different newly formed structures (reversible and stable) retain the tendency of enlargement of the pleural surface in all investigated periods. Simultaneous intercellular and transcellular transport, as an increase of the lysosomal system characterize the passing of the electron-dense material through the mesothelium. The early period (until 3rd day after EH) is characterized by more prominent mesothelial changes, involving activated cells. The initiation of the late period (on the 8th day after EH) begins with the appearance of lamellar bodies and newly formed elastic membrane. The following late changes (on the 15th day after EH) concern predominantly the components of the connective tissue layer, such as collagen accumulations and blood capillaries. The present data suggest that the alterations over the entire pleura are irregular and asynchronous, showing significant morphological differences in both pleura sheets, some of them are diffuse in character, the final ones appear to be stable and ensure incomplete pleural restoration. PMID- 15125041 TI - Decrease in laminin content and protein excretion rate after five sixths nephrectomy and low-dose irradiation in the rat. AB - The effect of low-dose irradiation on laminin distribution and urine protein excretion in the remnant rat kidney has been studied. The rat remnant kidney formed after 5/6 nephrectomy is an experimental model of chronic renal failure. In the remnant kidney, focal segmental glomerulosclerosis is developed characterized by focal or segmental sclerosis in glomeruli, alterations in the tubules and thickening of the glomerular basement membrane. Low dose irradiation has been presumed to suppress sclerotic processes. In this study 24 male Wistar rats were subdivided into the nephrectomized group, nephrectomized and irradiated groups (1 or 3 Grey), and healthy control group. Animals were sacrificed at 2, 4 and 8 weeks after beginning the experiment. Laminin immunohistochemical staining was found along the tubular and glomerular basement membranes in all experimental groups, but with varying intensity. Laminin content in the basement membranes was decreased in early stages (week 2), especially after irradiation followed by increase during the later stages with relatively high levels at the end of the experiment (week 8). Irradiation at a dose of 3 Grey decreased protein excretion compared to the nephrectomized rats at all stages, while 1 Grey dose was ineffective. Based on decreased proteinuria we conclude that moderate low-dose irradiation has beneficial effects on the rat remnant kidney and that laminin in basement membranes is probably not the most crucial component in regulating membrane permeability. PMID- 15125042 TI - 3D reconstruction of the cardiovascular and central nervous system of a human embryo Carnegie-stage 15--case report. AB - A human embryo at Carnegie stage 15 was serially sectioned and 3D computer aided reconstructions were made to demonstrate the cardiovascular system and cranial structures and to study developmental variations at this stage. The development of the heart and pharyngeal arteries was according to the existing literature. Differences were found in the development of the arterial circle of Willis and the central nervous system. The cranial venous system seemed to show great variability. Whereas the telencephalon was not developed according to the stage, the development of the hypophysis had occurred prior to stage 15. From the results we conclude that there are remarkable individual differences in embryological differentiation of structures which have to be taken into account during staging of human embryos. PMID- 15125043 TI - Age-dependent morphometrical changes in the thymus of male propranolol-treated rats. AB - In order to elucidate a putative role of neurally derived noradrenaline in the thymus development, and in maintenance of adult thymus structure, sexually immature male rats (21-day-old at the beginning of treatment) and young adult animals (75-day-old on the beginning of treatment) were treated with the non selective beta-adrenoceptor antagonist propranolol (0.40 mg/100 g BW/day, s.c.) for 15 consecutive days, and their thymuses were analyzed stereologically. The effects of beta-adrenoceptor blockade were much more pronounced in sexually immature than in adult rats. In immature propranolol-treated rats the thymus size and volumes of both the main compartments (cortex and medulla) were significantly decreased reflecting, at least partly, a reduction in the overall number of thymocytes. Furthermore, in both the cortical subcompartments (outer and deep cortex) the mean diameter of thymocytes was increased. However, in adult rats exposed to propranolol treatment, only the volume of interlobular connective tissue was enlarged, whereas in the outer part of the cortex the mean thymocyte diameter was increased. These results indicate that the lack of sympathetic input (via beta-adrenoceptors) during the prepubertal period of development diminishes the normal thymus growth and/or accelerates the thymic involution that starts at puberty, immediately after its maximum size is reached, while it is less significant for the maintenance of the thymus size and structure in adults. Additionally, they suggest that distinct cell types, as well as thymocyte subsets, are sensitive to lack of beta-adrenoceptor-mediated influences in sexually immature and adult rats. PMID- 15125044 TI - A simple fluorescence labeling method to visualize the three-dimensional arrangement of collagen fibers in the equine periodontal ligament. AB - In order to display the collagen-fiber arrangement in the equine periodontal ligament an inexpensive and easy staining procedure with fluorescein was applied to paraffin sections. After fluorescein labeling a section was suitable for successful examination with three special microscopical systems: a) fluorescence microscopy b) phase contrast microscopy and c) polarized light microscopy. Collagen fibers were clearly displayed as compact structures in the fluorescence microscope. This distinct feature of the fluorescent image generated an almost three-dimensional impression of the fiber arrangement. Phase contrast microscopy and polarized light microscopical investigations of the same section supplemented the findings with further structural details. This contributed to demonstration of the complex architecture of the PDL, i. e. the varying sizes of the fiber bundles, their specific spatial alignment, and the entheses to the dental cementum. PMID- 15125045 TI - An aberrant axillary artery penetrating the origin of the radial nerve from deep to superficial. AB - An aberrant axillary artery that penetrated the radial nerve from deep to superficial during its course, was observed. The brachial plexus in the present case was classified as the Adachi's C-type brachial plexus. Further, an accessory radial root existed, which was a nerve bundle branching from the deep aspect of the inferior trunk and communicating with the radial root from the posterior cord to form the radial nerve. The axillary artery went on along the lower border of the brachial plexus and passed between the radial root and the accessory radial root from deep to superficial at its third section. As the axillary artery penetrated the origin of the radial nerve from deep to superficial, it was judged to reach deep under the posterior cord, hence deeper than the brachial plexus. PMID- 15125046 TI - The Etruscan skulls of the Rostock anatomical collection--how do they compare with the skeletal findings of the first thousand years B. C.? AB - Seven Etruscan skulls were found in Corneto Tarquinia in the years 1881 and 1882 and were given as present to Rostock's anatomical collection in 1882. The origin of the Etruscans who were contemporary with the Celts is not yet clear; according to Herodotus they had emigrated from Lydia in Asia Minor to Italy. To fit the Etruscan skulls into an ethnological grid they were compared with skeletal remains of the first thousand years B.C. E. All skulls were found to be male; their age ranged from 20 to 60 years, with an average age of about thirty. A comparison of the median sagittal outlines of the Etruscan skulls and the contemporary Hallstatt-Celtic skulls from North Bavaria showed that the former were shorter and lower. Maximum skull length, minimum frontal breadth, ear bregma height, bizygomatical breadth and orbital breadth of the Etruscan skulls were statistically significantly less developed compared to Hallstatt-Celtics from North Bavaria. In comparison to other contemporary skeletal remains the Etruscan skulls had no similarities in common with Hallstatt-Celtic skulls from North Bavaria and Baden-Wurttemberg but rather with Hallstatt-Celtic skulls from Hallstatt in Austria. Compared to chronologically adjacent skeletal remains the Etruscan skulls did not show similarities with Early Bronze Age skulls from Moravia but with Latene-Celtic skulls from Manching in South Bavaria. Due to the similarities of the Etruscan skulls with some Celtic skulls from South Bavaria and Austria, it seems more likely that the Etruscans were original inhabitants of Etruria than immigrants. PMID- 15125047 TI - A quantitative study on the digital bones of cattle. AB - The bones of 64 digits from eight Holstein male cattle were studied quantitatively to determine whether any differences existed respective on the corresponding bones of the different digits. For this purpose, the greatest and abaxial greatest lengths, the smallest diaphysial breadths, the breadths of proximal and distal ends of the proximal and middle phalanges, the lengths of dorsal surfaces, the heights of extensor processes, the greatest diagonal lengths and the middle breadths of the soles from the distal phalanges were measured. Comparison showed that measurements between the right and left sides did not differ significantly. However, differences were found for almost all measurements between the forelimb and hindlimb. The proximal and middle phalanges were found to be shorter and broader in the forelimb and the broadness was more noticeable than the length. The distal phalanx of the medial forefoot had the greatest value in four measurements while the smallest values were found in the lateral hind foot. The results indicated that the morphometry of the digital bones could be important from both phylogenetic and clinical aspects. PMID- 15125048 TI - Cribra orbitalia visualized in computed tomography. AB - Cribra orbitalia are sieve-like lesions of the orbital roof. While common in historic skulls, they have long been absent in those examined in the last decades. Only recently we found low-grade cribra orbitalia in some contemporary cases. Though of unknown origin, this lesion is often attributed to anemia and deficiency diseases. It is theoretically possible to visualize cribra orbitalia in living subjects by computed tomography and thus study their etiology. The aim of our study was to investigate the possibilities of computed tomography for visualizing cribra orbitalia. We used multislice computed tomography (MSCT) in the spiral and sequential mode to image medium-grade cribra orbitalia of moderate severity in a human skull. Virtual endoscopic and 3-D images were produced by post-processing. The best results were obtained by the sequential mode and 3-D reconstruction. Given a thin slice thickness and a wide slice angle between slice plane and the orbital roof, the threshold level seems to be the most important factor influencing realistic reproduction and should be finely adjusted according to bone density. Clinical research may now begin by examining relevant patients undergoing CT. PMID- 15125049 TI - Communication between the buccal nerve (V) and facial nerve (VII) in the human face. AB - Terminal arrangements of communicating branches between the buccal nerve (V) and the facial nerve (VII) have yet to be precisely determined. To clarify distributions and relationships to facial muscles, detailed morphological examination of the two nerves was performed in the buccal region. The facial skin and underlying tissues of three cadavers were removed en bloc from the surface of the skulls and dissected from outside and inside. Arrangements of the facial muscles, nerves, and associated structures were observed. In all specimens, the communicating buccal nerve (CBN) was detected, largely covering the buccal region. The CBN gave off multidirectional twigs around the facial vein, some of which reached the anterior part of the zygomaticus major muscle. Several twigs of the CBN joined proximally with the zygomatic and buccal branches of the facial nerve. Ramified junctions that interconnected the lower zygomatic and upper buccal branches of the facial nerve were observed near sites where the CBN joined. Anterior twigs of the CBN supplied the longitudinal area lateral to the mouth, where many muscles converged. This study presents a precise morphological pattern of the CBN, suggesting functional contribution of the CBN to control of orofacial movements. PMID- 15125050 TI - The scanning electron microscopic study of lingual papillae in the silver fox (Vulpes vulpes fulva, Desmarest, 1820). AB - The tongues of adult silver foxes were studied using scanning electron microscopy. Five types of lingual papillae were found on the dorsal surface of the tongue. The most numerous papillae were filiform papillae covering the apex and body of the tongue. The filiform papillae on the anterior part of the tongue are divided into 1 main and 10-12 accessory processes. In the posterior part of the body of the tongue the number of accessory processes is reduced. Fungiform papillae are located between the filiform papillae. A cluster of 12 large fungiform papillae was found on the apex of the tongue. Conical papillae are located in the area of the vallate papillae and cover the posterior part of the root of the tongue. Their size increases towards the pharynx, where they are distributed more sparsely. In the silver foxes there were two pairs of vallate papillae. The wall surrounding each papilla and its gustatory trench forms partly connected 6-8 conical papillae. The foliate papillae on both margins of the tongue body are small and consist of 4-5 laminae. The distribution and type of lingual papillae found in the silver fox are similar to those in the other species belonging to the family Canidae. PMID- 15125051 TI - Variability in the human m. spinalis capitis and cervicis: frequencies and definitions. AB - The spinalis muscle is defined as the medial component of the erector spinae muscle group, and is typically subdivided into three regional components: m. spinalis thoracis, m. spinalis cervicis and m. spinalis capitis. Modern authorities, however, differ on the morphology of the cervicis and capitis portions and many claim that these regional distinctions do not commonly exist. This article reports on the validity of that claim. Three morphological conditions are identified for both m. spinalis capitis and m. spinalis cervicis. The distinct condition has the muscle persisting as an independent muscular unit. The blended condition occurs when the spinalis muscle fibers blend with those of the semispinalis muscle. Finally, there are occasions in which no evidence of spinalis muscle fibers can be found. Frequency estimates are provided for each of these morphological conditions. Results of statistical analyses indicate that there are no relationships among muscle varieties and the sex of the individual or the side of the body where the muscle was found. PMID- 15125052 TI - Cortical coding. AB - Physiological photic and sound stimulation generated almost synchronous summated cortical evoked peak interval patterns over all regions of the head studied in man. Although the peak interval synchrony in all regions was obvious, it was observed that the peak amplitudes, and even phase relations varied over the different brain regions. The almost invariant peak time interval patterns for the input modalities studied, suggested a simple, generalized direct cortical coding that signaled immediate environmental change to the entire brain. PMID- 15125053 TI - Tremor in Parkinson's disease: 24-hr monitoring with calibrated accelerometry. AB - Monitoring systems enable the long-term registration of tremor in patients with Parkinson's disease This method is useful in the objective measurement of tremor during the course of treatments. Indeed, the symptoms of tremor as well as the aggravating and attenuating influences can be observed under real-life conditions. The methodology of data recording and analysis, described in previous investigations, was extended to automatically detect body position and certain movement patterns with calibrated 4-channel accelerometry. The main purpose of the present investigation was to apply this refined and extended methodology to patients in a clinical rehabilitation program, and to examine its practability with respect to the results of the treatment and the patients' compliance. The methodology was tested on 30 patients (17 male, 13 female) with Parkinson's disease. The mean age was 64.8 years (s = 8.9). The Hoehn-Yahr index ranged from 1 to 3 (m = 2.3, s = 0.7) and the overall UPDRS scale between 10 and 74 (m = 42.9, s = 18.1). The data recording included: (1) the registration of tremor under standardised conditions of rest and postural tremor test with and without distraction; (2) a standard protocol to obtain reference values for body position and movement; and (3) the 24-hr monitoring. 21 patients could be recorded a second time, on average 18 days after the first recording. Between the two registrations, patients received individually tailored drug treatment supplemented with specific activating physiotherapy, ergotherapy measures, and individual psychotherapeutic counseling. Changes between first and second recording were evident for the three tremor variables, but significant only for the 24-hr ambulatory monitoring. The between and within-subjects correlations of the tremor variables were rather low except the correlations between occurrence and amplitude (between-subjects. 87; within-subjects. 67). Conditions of rest and postural tremor test showed a correlation with corresponding segments of the ambulatory monitoring of about. 50 for the tremor occurrence. The best prediction of the day-time monitoring was made by the tremor tests with distraction, whereas the night segment was best predicted by the standard protocol. PMID- 15125054 TI - Excitability properties of normal and demyelinated human motor nerve axons. AB - The strength-duration time constants and rheobase currents, which provide an indirect indication of the axonal properties are calculated in two cases of stimulation, using our previous double cable models of normal and demyelinated human motor fibres. The time constants and rheobases are defined as nodal when the case of point fibre polarization (intracellular current application at the first node) is used for their calculations, whereas the time constants and rheobases are defined as internodal when the case of periodic kind of uniform fibre polarization (simultaneous intracellular current application at each axon segment) is used. Four fibre demyelinations (termed as paranodal focal 1 systematic and internodalfocal 1 systematic demyelinations) are studied. For both investigating cases of current application, the stimulus duration is increased in 0.025-ms steps from 0.025-ms to 1-ms and the strength-duration and charge duration curves are plotted for the axons. The strength-duration time constants are calculatedfrom the curve-fitting equation for the resulting charge-duration curves. The results are consistent with the interpretation that the time constants depend not only on the types of the demyelinated axon, but on the methods of fibre stimulation. The strength-duration time constants (nodal 1 internodal) are almost the same for the normal axons and focally demyelinated axons, however, they are shorter for the paranodally systematically demyelinated axons, and longer for the internodally systematically demyelinated axons. For all investigated cases, the internodal time constants are greater than the nodal time constants and there is an inverse relationship between the time constants and rheobase currents. PMID- 15125055 TI - Comparison of linear regression and probit analysis for detecting H-reflex threshold in individuals with and without spinal cord injury. AB - A major challenge to understanding spinal reflex organization in health and disease is identifying sensitive measures of reflex excitability. The purpose of this study was to determine whether linear regression or probit analysis techniques are more sensitive for detecting H-reflex and M-wave threshold and for identifying differences in H-reflex threshold in individuals with and without spinal cord injury (SCI). Soleus H-reflex recruitment curves were generated in 9 individuals with SCI and 20 able-bodied individuals. H-reflex and M-wave threshold was estimated using three different methods, two that used linear regression of H-reflex peak-to-peak amplitude and one that used probit analysis of quantal H-reflexes. Results indicate that in both groups all three techniques were equally sensitive for detecting H-reflex but not M-wave threshold. When H reflex threshold was normalized to M-wave threshold, different techniques provided different estimates of H-reflex threshold. However, between-group differences (SCI vs. able-body) in H-reflex and M-wave threshold were not affected by the measurement techniques that were compared in this study. We conclude that these techniques provide equally sensitive estimates of H-reflex but not M-wave threshold in persons with and without SCI. Hence, caution should be used when interpreting normalized and non-normalized values of H-reflex threshold. PMID- 15125056 TI - Clinical, electrophysiological and magnetic resonance imaging findings in carpal tunnel syndrome. AB - OBJECTIVE: To assess magnetic resonance imaging (MRI) findings in carpal tunnel syndrome (CTS) and to compare them with electrophysiological findings. METHODS: Routine motor and sensory nerve conduction examinations and needle EMG were performed in 42 hands of 22 patients, who were clinically diagnosed as having CTS in at least one wrist. RESULTS: Of 29 wrists with clinically and electrophysiologically confirmed CTS, MRI could detect abnormality in 18 wrists (62%). Median nerve was found to be abnormal in MRI in 1 of 2 wrists with suspected clinical symptoms and proven CTS by electrophysiological examination. MRI was abnormal in 1 of 4 wrists with normal clinical and electrophysiological examination. MRI was abnormal in 46, 7% of wrists with mild CTS, in 61.6% of moderate CTS and in 100% of severe CTS. Volar bulging of the flexor retinaculum was detected in a single wrist with severe CTS. Enlargement of median nerve was observed in 3 of 5 severe CTS. CONCLUSION: MRI could be useful in the diagnosis of unproven cases in CTS. It also provides anatomical information that correlate well with electrophysiological findings in regard of the severity of median nerve compression. PMID- 15125057 TI - Neurogenic vestibular evoked potentials using a tone pip auditory stimulus. AB - OBJECTIVES: To obtain neurogenic vestibular evoked potentials (NVESTEPs) with surface scalp recording using a tone pip auditory stimulus. METHODS: Fourteen neurologically normal volunteers (Age range 26-45 years, 10 females and 4 males), and two patients with sensorineural hearing loss and possible multiple sclerosis respectively, were examined. Two channel recordings were obtained, the first channel being P3 referred to Fpz, and the second channel being P4 referred to Fpz. A 1 kHz tone pip stimulus with two cycles was delivered via headphones monoaurally with contralateral masking noise. RESULTS: A consistent negative wave with a mean absolute latency of 4.72 msec was obtained, which we have named N5. 25% of the ears tested had better responses at the ipsilateral parietal electrode. In the patient with bilateral sensorineural hearing loss, NVESTEPs was present, suggesting that the NVESTEP is not a cochlear response. In the patient with possible multiple sclerosis, an abnormal NVESTEP response and a normal BAEP response were found. CONCLUSION: Use of a tone-pip rather than a click auditory stimulus allows a lower click intensity to be used in the production of NVESTEP responses, leads to a shorter testing time, and is therefore more comfortable for the patient. This study adds to our impression that the NVESTEP may be a physiological response that can be used to assess the vestibular system and is different from the BAEP response. Further testing in patients with symptoms of dizziness and with disorders specific for the vestibular nerve is required. PMID- 15125058 TI - Changes of reflex size in upper limbs using wrist splint in hemiplegic patients. AB - We evaluated the effect of prolonged wrist extension on H reflex in the flexor carpi radialis (FCR) muscle and tendon jerk (T) reflex in the biceps brachii (BB) muscle of 17 chronic hemiplegic patients. H reflex of the FCR and T reflex of the BB were assessed every 5 minutes within 20 minutes during prolonged wrist extension and post-20 minutes after the extension. As a result, H reflex in the FCR was reduced by passive wrist stretch in 82% of the spastic limbs. The effect was larger in the higher spastic group. In 45% of the spastic limbs, T reflex in the BB also was reduced by passive wrist stretch. The inhibitory effects had a tendency to strengthen in accordance with the grade of muscle tone. We considered from these results, prolonged wrist extension generated inhibitory projections via probably group II afferents of the FCR in the homonym and in the transjoint in spastic limbs. PMID- 15125059 TI - Bilateral anterior interosseous neuropathy following surgery: a case report. AB - The authors present a patient who developed bilateral anterior interosseous neuropathy following an abdominal surgical procedure. This case is of particular interest because while anterior interosseous nerve involvement may be seen with Parsonage-Turner Syndrome, the bilateral involvement is quite rare. The electrodiagnostic findings are presented and the literature reviewed. PMID- 15125061 TI - Porcine genomics delivers new tools and results: this little piggy did more than just go to market. AB - The past decade has yielded new tools for pig geneticists and breeders thanks to the considerable developments resulting from efforts to map the pig genome. The pig genetic linkage map now has nearly 5000 loci including several hundred genes, microsatellites and amplified fragment length polymorphisms (AFLP) markers. Using tools that include somatic cell hybrid panels and radiation hybrid panels, the physical genetic map is also growing rapidly and has over 4000 genes and markers. Scientists using both exotic and commercial breeds for quantitative trait loci (QTL) scans and candidate gene analyses have identified a number of important chromosomal regions and individual genes associated with growth rate, leanness, feed intake, meat quality, litter size and disease resistance. Using marker assisted selection (MAS) the commercial pig industry is actively incorporating these gene markers and traditional performance information to improve traits of economic importance in pig production. Researchers now have novel tools including pig gene arrays and advanced bioinformatics that are being exploited to find new candidate genes and to advance the understanding of gene function in the pig. Sequencing of the pig genome has been initiated and further sequencing is now being considered. Advances in pig genomics and directions for future research and the implications to both the pig industry and human health are reviewed. PMID- 15125060 TI - Combined median and medial antebrachial cutaneous neuropathies: an upper-arm neurovascular syndrome. AB - The neurovascular bundle anatomy in the upper arm displays changing relationships of nerve and vascular structures along short segments. Fibrous tissues segregate these elements into enclosed compartments allowing for specific patterns of injury. We report a patient with a iatrogenic brachial artery injury in this region who featured combined median and MAC neuropathies, which were consistent with complete axonotmesis on neurophysiological assessment. Increased intracompartmental pressure may have led to nerve injury either thorough an ischemic mechanism or to focal compression. Recognition of this unusual pattern of nerve damage is important, since injury can be accurately localized to the midportion of the neurovascular compartment in the upper arm. PMID- 15125062 TI - Mutation accumulation and the effect of copia insertions in Drosophila melanogaster. AB - Repeated efforts to estimate the genomic deleterious mutation rate per generation (U) in Drosophila melanogaster have yielded inconsistent estimates ranging from 0.01 to nearly 1. We carried out a mutation-accumulation experiment with a cryopreserved control population in hopes of resolving some of the uncertainties raised by these estimates. Mutation accumulation (MA) was carried out by brother sister mating of 150 sublines derived from two inbred lines. Fitness was measured under conditions chosen to mimic the ancestral laboratory environment of these genotypes. We monitored the insertions of a transposable element, copia, that proved to accumulate at the unusually high rate of 0.24 per genome per generation in one of our MA lines. Mutational variance in fitness increased at a rate consistent with previous studies, yielding a mutational coefficient of variation greater than 3%. The performance of the cryopreserved control relative to the MA lines was inconsistent, so estimates of mutation rate by the Bateman-Mukai method are suspect. Taken at face value, these data suggest a modest decline in fitness of about 0.3% per generation. The element number of copia was a significant predictor of fitness within generations; on average, insertions caused a 0.76% loss in fitness, although the confidence limits on this estimate are wide. PMID- 15125064 TI - Population subdivision and the Hudson-Kreitman-Aguade test: testing for deviations from the neutral model in organelle genomes. AB - The Hudson-Kreitman-Aguade (HKA) test is based on the prediction from the neutral theory that levels of polymorphism within a species and the divergence between two closely related species should be correlated. Population subdivision has been shown to alter both the amounts of polymorphism segregating within species and the rate of divergence between species, meaning that genomic regions with different population structures also differ in their divergence to polymorphism ratios. Population subdivision may hence hamper the utility of the HKA test for detecting deviations from the standard neutral model, especially for organelle genomes that often have different patterns of population structure compared with nuclear genes. In this paper, I show that population subdivision inflates the number of instances where the HKA test detects deviations from the neutral model. Using coalescent simulations I show that this bias is most apparent when population subdivision is strong and differs substantially between the loci included. However, if divergence time is large and population structure substantial even changes in the levels of polymorphism and divergence associated with differences in the effective population size between two loci is enough to substantially alter the number of significant outcomes of the HKA test. A dataset on cytoplasmic diversity in Sileine vulgaris and S. latifolia (Ingvarsson & Taylor, 2002) is also reanalysed. The previous study had shown a marked excess of intraspecific polymorphism in both species. However, when effects of population subdivision were removed, ad hoc, levels of intraspecific polymorphism were no longer significantly different from neutral expectations, suggesting that population subdivision contributed to the observed excess of intraspecific polymorphism seen in both species of Silene. PMID- 15125063 TI - Isolation and characterization of dinucleotide repeat microsatellites in Drosophila ananassae. AB - Drosophila ananassae is a cosmopolitan species with a geographic range throughout most of the tropical and subtropical regions of the world. Previous studies of DNA sequence polymorphism in three genes has shown evidence of selection affecting broad expanses of the genome in regions with low rates of recombination in geographically local populations in and around India. The studies suggest that extensive physical and genetic maps based on molecular markers, and detailed studies of population structure may provide insight into the degree to which natural selection affects DNA sequence polymorphism across broad regions of chromosomes. We have isolated 85 dinucleotide repeat microsatellite sequences and developed assay conditions for genotyping using PCR. The dinucleotide repeats we isolated are shorter, on average, than those isolated in many other Drosophila species. Levels of genetic variation are high, comparable to Drosophila melanogaster. The levels of variation indicate the effective population size of an Indonesian population of D. ananassae is 58,692 (infinite allele model) and 217,284 (stepwise mutation model), similar to estimates of effective population size for D. melanogaster calculated using dinucleotide repeat microsatellites. The data also show that the Indonesian population is in a rapid expansion phase. Cross-species amplification of the microsatellites in 11 species from the Ananassae, Elegans, Eugracilis and Ficusphila subgroups indicates that the loci may be useful for studies of the sister species, D. pallidosa, but will have limited use for more distantly related species. PMID- 15125065 TI - A comparison between methods for linkage disequilibrium fine mapping of quantitative trait loci. AB - We present a maximum likelihood method for mapping quantitative trait loci that uses linkage disequilibrium information from single and multiple markers. We made paired comparisons between analyses using a single marker, two markers and six markers. We also compared the method to single marker regression analysis under several scenarios using simulated data. In general, our method outperformed regression (smaller mean square error and confidence intervals of location estimate) for quantitative trait loci with dominance effects. In addition, the method provides estimates of the frequency and additive and dominance effects of the quantitative trait locus. PMID- 15125066 TI - An improved formulation of marker heterozygosity in recurrent selection and backcross schemes. AB - This report presents a theoretical formulation for predicting heterozygosity of a putative marker locus linked to two quantitative trait loci (QTL) in a recurrent selection and backcross (RSB) scheme. Since the heterozygosity at any given marker locus maintained in such a breeding programme reflects its map location relative to QTL, the present study develops the theoretical analysis of the QTL mapping method that recently appeared in the literature. The formulae take into account selection, recombination and finite population size during the multiple generation breeding scheme. The single-marker and two-QTL model was compared numerically with the model involving two linked marker loci and two QTL. Without recombination interference, the two models predict the same expected heterozygosity at the linked marker loci, indicating that the model is valid for predicting marker heterozygosity maintained at any loci in an RSB breeding scheme. The formulation is demonstrated numerically for several RSB schemes and its implications in developing a likelihood-based statistical framework for modeling the RSB experiments are discussed. PMID- 15125067 TI - Mendelian sampling terms as a selective advantage in optimum breeding schemes with restrictions on the rate of inbreeding. AB - Quadratic indices are a general approach for the joint management of genetic gain and inbreeding in artificial selection programmes. They provide the optimal contributions that selection candidates should have to obtain the maximum gain when the rate of inbreeding is constrained to a predefined value. This study shows that, when using quadratic indices, the selective advantage is a function of the Mendelian sampling terms. That is, at all times, contributions of selected candidates are allocated according to the best available information about their Mendelian sampling terms (i.e. about their superiority over their parental average) and not on their breeding values. By contrast, under standard truncation selection, both estimated breeding values and Mendelian sampling terms play a major role in determining contributions. A measure of the effectiveness of using genetic variation to achieve genetic gain is presented and benchmark values of 0.92 for quadratic optimisation and 0.5 for truncation selection are found for a rate of inbreeding of 0.01 and a heritability of 0.25. PMID- 15125068 TI - [Waldenstrom macroglobulinemia]. PMID- 15125069 TI - [Dementia assessment in the Memory Clinic St. Gallen: do further diagnostic work up or therapy lead to a change of diagnosis?]. AB - We evaluated 143 patients referred to the Memory Clinic St. Gallen for ambulatory dementia assessment between 1995 and 1999. We assessed demographic data, medical results and follow-up data over a mean period of 16 months. 63% of the patients suffered from dementia, 14% had a mild cognitive impairment, 1% had a preexisting debility and 22% had no cognitive deficit. The cause of dementia was Alzheimer's disease in 40% of the patients. Vascular dementia (2.8%) and reversible forms of dementia (2.1%) were underrepresented. In 63 out of 143 patients we examined, if further diagnostic work up (CT or MRI, laboratory testing etc.) or empirical medication by the family physicians led to a change of the initial diagnosis. Follow-up findings confirmed the initial diagnosis in 87.3% of cases. Based on these results we conclude that expensive diagnostic procedures are not necessary in most patients referred for the ambulatory assessment of dementia. PMID- 15125070 TI - [Langerhans cell histiocytoses: 50 years to histiocytosis X]. AB - Histiocytoses encompass a heterogeneous group of histiocytic disorders that may be separated in a group of variable biological behaviour and in a clearly malignant group. The histopathologic diagnosis of all types of histiocytoses is always mandatory. The 50-year-old Histiocytosis X compiles a subgroup of different biological behaviour, nowadays called Langerhans cell histiocytoses- the subject of the review. It has a wide range of manifestations: self-healing cutaneous, benign solitary and malignant systemic manifestations require the staging in order to evaluate a possibly necessary therapy. Chemotherapy should be performed within international protocols. PMID- 15125071 TI - [Polyneuropathy seropositive for anti-Hu antibodies]. PMID- 15125072 TI - [Rediscovery of monastery plants]. PMID- 15125073 TI - [Headache]. PMID- 15125074 TI - [Review: functional foods and ingredients derived from milk]. AB - The objective of this paper is to review the main research works related to functional foods and ingredients derived from milk. Research in functional foods has increased during last twelve years with the intention of increasing life expectancy and improving human health conditions. Probiotics, prebiotics, bioactive peptides or proteins, dietetic fibers and fatty acids, as well as the addition of fitochemical compounds in dairy products and a record of some allergic compounds are also discussed. The demand of this kind of products is increasing due to intense advertising campaigns posted in many countries. Basically, these campaigns promise better health and/or the prevention of certain illnesses. Milk contains diverse constituents with physiological functionality, which might change the traditional view point that we have about drugs. The topic of functional foods in general, and specifically that from milk and dairy products, has still not been completely exploited, and in the future it will be found that the best work has not been carried out in this area. PMID- 15125075 TI - [Germinated or fermented legumes: food or ingredients of functional food]. AB - Epidemiological research has shown a positive association between certain diseases and dietary intake of food components found in fruits, grains, legumes, fish oil among others. Food that may provide a health benefit beyond the traditional nutrients that it contains, are named functional food. In addition to the varied nutrients, legumes contain compounds such as polyphenols, soluble fiber, alpha-galactosides and isoflavones which confer propierties of functional foods. Do to the cuse of flatus production in some people, long cooking periods, or anti-nutritional factors, legume consumption levels are limited. In this review, germination and fermentation processes will be presented as alternatives that are able to reduce or inactivate anti-nutritional factors, preserve and even improve the content of the isoflavones, or better the potencial of the legumes as functional food or as ingredients for the formulation of functional foods. PMID- 15125076 TI - [Vitamin A deficiency and clinical associations: a review]. AB - An important focus of attention in Public Health has been micronutrient deficiency in human being because of the enhanced vulnerability of individuals to the effects of micronutrient deficiency or imbalance. Among all micronutrients deficiencies, vitamin A has been one of the most important public health problems, affecting a large percentage of people in developing countries. Vitamin A is particularly important for its role in the process of organism defense against infections. This article reviews comprehensively vitamin A metabolism, it highlights hypovitaminosis A relationship with pregnancy and human imunodeficiency, showing its repercussions in several clinical conditions. PMID- 15125077 TI - [Vitamin A status in pregnant adolescents of low socioeconomic income]. AB - Vitamin A is an essential micronutrient for growth, especially in highly proliferative and development stages as in pregnancy. When maternal vitamin A is deficient, fetal demands does not allow to maintain maternal reserves and subclinical deficiency appears. Due to the fact that pregnancy is a period of vulnerability to nutritional deficiencies, 75 pregnant adolescent (16.4 +/- 1.2 years old), were studied along prenatal control visits during pregnancy for vitamin A status. Data on serum retinol was obtained in each trimester through fasting blood collection, and determination by high pressure liquid chromatography. Two-24 hour recalls during each trimester were used to assess dietary intake. Statistical analysis (descriptive, frequency distribution, paired t-test and Fisher's exact test) were performed. Mean serum retinol values decreased significantly (p < 0.05) along trimesters. There was not vitamin A deficiency by serum retinol indicator at any trimester, but at third trimester, 30.3% of the women showed marginal serum retinol values (20-30 mg/gL). Dietary intake, adjusted for recommendations for each trimester, increased significantly as pregnancy continued. These findings indicate that even though serum retinol (vitamin A) deficiency was not present at any time during pregnancy in these adolescents, risk of deficiency increased toward the end; and that the increase of vitamin A intake at the end of pregnancy does not change the decline in serum retinol. Monitoring and nutritional intervention of this vulnerable group is recommended in order to promote a healthy mother and newborn. PMID- 15125078 TI - [Relationship between maternal anthropometry and weight gain with birth weight, and risks of low birth weight, small for gestational age and prematurity at an urban population of Buenos Aires, Argentina]. AB - To asses the relationship between body mass index and net weight gain during pregnancy with birth weight and the risks of low birth weight, small for date and prematurity 9613 records from Sarda's Perinatal Database between 1994-1995 were reviewed. Exclusion criteria were fetal death, twin pregnancy, congenital malformations, lack of prenatal visits and lack of preconceptional weight and height. 9.6% of mothers and 15% of adolescents presented with low preconceptional BMI (median: 24.8 +/- 4.3 kg/m2); in contrast, 28% were overweight and obese. Net weight gain (median 9.25 +/- 4.9 kg) accounted for 16% of previous weight and was higher with lower BMI (p = 0.001). Birth weight (median 3375 +/- 467 g) decreased with lower BMI (p = 0.001) and the risks of low birth weight (p < 0.05), small for date (p < 0.05), and prematurity (p = 0.05) was independently associated with BMI, and increased (p < 0.001) when lower the net weight gain was. The best predictors for low birth weight, small for date and prematurity risks were low preconceptional weight (40-51 kg) (adjusted OR 1.72; [95%CI 1.48-1.95], 2.12 [1.82-2.41] and 1.46 [1.12-1.79] respectively). Net weight gain and several predictive variables did not explain more than 10.8% of the variability of birth weight. Preconceptional weight should have important implications for the design of future nutritional strategies at a poblational level, especially for adolescents. PMID- 15125079 TI - [Excess of weight and their relationship with high blood pressure in schoolchildren and adolescents of Medellin, Colombia]. AB - A cross sectional study was carried out in order to investigate the association between excess weight based on body mass index (> or = p85 Must scales), and high diastolic and sistolic blood pressure (> or = p90 Task Force Blood Pressure Control in Children scales). The sample consisted of 1253 male and 1358 female, aged 6 -18 years in the city of Medellin, Colombia. Excess weight was observed in 14.3% of males and 13.7% of females, with higher prevalence at 6 to 9 years old in any gender. Diastolic high blood values were significantly higher in males (4.9%) when compared to females (2.9%) (p = 0.007), and high sistolic values were found similarly in males (1.6%) and females (1.0%) (p = 0.203). This study confirmed the statistically significant association between high body mass index (> or = p85) and high sistolic (RP 4.04; IC 2.03-8.04) and diastolic (RP 3.44; IC 2.32-5.09) blood pressure. PMID- 15125080 TI - [Microbiological quality and effect of washing and disinfection of pre-cut Chilean vegetables]. AB - Actually it is possible to find a great offer of pre-cut vegetables in the Chilean market. These products present certain advantages, such as saving time in their preparation at home. The microbiological quality of some of these vegetables, pre-cut celery and pre-cut cabbage was assayed. Two different types of the products were studied. The traditional pre-cut vegetable and another one, which is also pre-cut but it is labeled as "ready-to-eat" (minimally processed). The last one could be consumed without previous washing or disinfecting. The assayed microbiological parameters were: total plate count, Enterobacteriaceae count, total coliforms and fecal coliforms most probable number, investigation of Escherichia coli and absence or presence of Salmonella. The effect of washing and washing and disinfecting on the natural microflora was also carried out. A disinfectant product for home use was obtained from the market. When comparing the obtained results for the two vegetables from both types, a high level of total plate count, Enterobacteriaceae count and total coliforms most probable number was observed. Fecal coliforms were detected only in samples of the traditional type in both vegetables. In each case E. coli was identified. No samples showed presence of Salmonella. Washing and washing and disinfecting effect was low. The maximum reduction of the present microflora of both vegetables was only up to 2 logarithm cycles. PMID- 15125081 TI - [Presence of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella spp. in food from animal origin in Costa Rica]. AB - The presence of Escherichia coli O157:H7, Listeria monocytogenes and Salmonella spp was analyzed in two kinds of food from animal origin in Costa Rica. 100 samples of non pasteurized milk, from the principal producing zones of the country, and 100 samples of chicken giblets, purchased in retail markets, were analyzed according to the methodology described by Food and Drug Administration, 1995. Escherichia coli O157:H7 was analyzed in both kinds of food, while L. monocytogenes was evaluated in raw milk and Salmonella spp. in chicken giblets. Five strains of E. coli O157:H7 were isolated, three of them coming from chicken giblets and the other two from raw milk. 15% positivity for Salmonella spp. was found in chicken giblet samples and 3% positivity for L. monocytogenes in raw milk samples. The results obtained show the importance of the adequate processing of food from animal origin in order to decrease the potential transmission of pathogenic agents. The introduction of Hazard Analysis Critical Control Points system (HACCP) and Good Manufacturing practices in food industry keep on being the principal control measures and inocuity warranty. PMID- 15125082 TI - [Evaluation of the AOAC 985.29 enzimic gravimetric method for determination of dietary fiber in oat and corn grains]. AB - The precision attributes and use of the enzymatic-gravimetric method of Prosky et al. (1992) (AOAC 985.29) were evaluated using corn (BR 5202 Pampa) and oat (UFRGS 15) samples. The effect of laboratory batches carried out in different days were evaluated in six laboratory batches, using for each material one duplicate for total fiber (FT) determination, one duplicate for insoluble fiber (FI) determination and blank ones for FT and for FI (both in duplicate). In order to characterize repetitive aspects, five other FT and FI determinations added to each sample were evaluated, summing up 11 data. The low coefficients of variation in the first six batches were considered acceptable as an expression of expected total intralaboratory variation. The repetitive of the method was considered good for FT determinations (CVs < 10%). However, in the FI determination a high frequency of negative values of ash and blanks was found, impairing the repetitive aspects evaluation. The magnitude of the total gravimetric corrections varies with the kind of the sample and is especially influenced by the protein content. PMID- 15125083 TI - [Functional properties of Sphagnum magellanicum fiber and its direct use in formulation of bakery products]. AB - Characterization of functional properties of Sphagnum magellanicum fiber were investigated. Water absortion (WAC) and water retention (WRC) capacities, swelling capacity (SC); organic molecule absortion capacity (OMAC) and cationic interchange capacity (CIC) were evaluated, as well as its incorporation as fiber source to bakery produts. Diferent particles sizes were selected to evaluate their effects on the functional properties of moss fiber: T1(1.4 mm);T2(1.0 mm); T3(0.42 mm); T4(0.18 mm). Best results of CAA, CRA; SC and OMRC were obtained with T3, whereas best values of CIC were attained with T1. An optimized formulation of fiber enriched bread was developed analizing simultaneously the effect of four independent variables (yeast, moss fiber, fluffy agent and shortening) on the sensory quality of products. Shelf life studies were carried out by storing samples of fiber enriched breads at 20 degrees C and 6 degrees C. At the end of the study, refrigerated samples showed better sensory quality stability. PMID- 15125084 TI - [Iodine content of cattle milk from mountain and coast from Peru]. AB - With the objective to establish the cause-effect relationship between a geological area and the iodine content in cattle milk, and to estimate the contribution of milk consumption to the dietary iodine intake, the iodine content in cattle milk from the sierra and the coastal regions of Peru was determined. Milk samples were collected of cows from the three main productive zones of Peru, 62 in Cajamarca, 42 in Arequipa, both in the sierra, and 27 in Lima at the coast. The measurement of iodine was made by the method of Zak, based on the Sandell- Kolthoff reaction. The median values obtained were 24 microg/L in Cajamarca, 34 microg/L in Arequipa, and 170 microg/L in Lima. The median value in the sierra, 26 microg/L, was significantly lower than the one found in the coast. Moreover, while in the former 81% of individual values were below 50 microg/L, in the latter, on the contrary, 77% were above 80 microg/L. These results confirm that the iodine content in cattle milk is related to ecological factors. At the same time, they demonstrate that the iodine content in milk from the sierra is six times lower than in milk from the coast, and also that its consumption does not contribute significantly to satisfy the human physiological requirements of iodine in that zone. PMID- 15125085 TI - [Dietary fiber in fruits cultivated in Chile]. AB - The objective of this study was to determine the total, soluble and insoluble dietary fiber contents in fruits produced in Chile. The analyses were conducted in the fruits as eaten. Thirty eight fruits obtained from local markets and the Instituto de Investigaciones Agropecuarias (INIA) were studied. Water and total, soluble and insoluble dietary fiber were determined in the edible part of each sample. Moisture ranged between 73.1 and 92.4 g/100g. Average (+/- sd) total, soluble and insoluble contents, expresed as g/100 g on wet basis were: 2.41 +/- 1.26, range 0.30 - 5.62; 0.73 +/- 0.50 range 0.07 -2.26 and 1.68 +/- 0.89, range 0.15 -3.36 respectively. On dry weight basis total fiber concentration was 16.5 +/- 8.8, with a proportion of 69.2 +/- 11.2% and 30.8 +/- 11.2% of the insoluble and soluble form, respectively. Dietary fiber supply ranged between 0.6 to 8.4 g in the medium serving sizes. The ratios insoluble fiber/ crude fiber and total fiber/crude fiber did not present constant results. Values ranged between 1.1 and 4.9 (mean 2.5 +/- 1.1) in the former, and from 1.6 to 8.0 (mean 3.6 +/- 1.7) in the latter. It is concluded that both soluble and insoluble fiber vary widely among vegetables fruits produced in Chile. This study provides information on the fiber composition of fruits. Such information may help to choose them according to these variables in order to be used in the prevention or treatment of selected pathologies. PMID- 15125086 TI - [Chemical composition and digestibility of mote]. AB - White and yellow maize grains (Zea mays amylacea, capia), polished by means of heat treatment with lime and ashes were analyzed (maize for mote); the chemical composition and digestibility in vitro of these grains cooked in boiling water were studied (mote), they were obtained under standardized conditions (laboratory) to compare them with the ones purchased at stores. The composition of nutrients was similar in polished grains, standing out the white ones from laboratory with a protein and sodium content of 9.95 100 g/100 g and 80.33 mg/100 g in the ones processed with ashes respectively and calcium (979.70 mg/100 g) in the ones treated with lime. Motes purchased at stores were analyzed and turned out to be chemically similar to the ones from laboratory. In the white motes with ashes of this last group, values of sodium, iron and potassium (28,02; 7,95 and 75.64 mg/100 g respectively) were higher than those that received treatment with lime (white and yellow), which presented a high calcium content (464.97 and 430.79 mg/100 g respectively). The digestibility was superior in the samples with lime, 79% in the purchased ones and 80% in the standardized ones. PMID- 15125087 TI - Unsafe abortion and maternal mortality: is Africa prepared to face the reality? PMID- 15125088 TI - How research can affect policy and programme advocacy: example from a three country study on abortion complications in sub-Saharan Africa. AB - OBJECTIVE: To provide a basis for continued policy dialogue and reform to address the problem of death due to abortion complications among the ECSA Health Community countries. The anticipated short-term outcome of this study was increased awareness among African health officials about the problem of incomplete abortion in SSA, especially CRHCS/ECSA countries. DESIGN: The study undertaken in 1993/1994, involved primary data collection of abortion-related morbidity and mortality statistics, the cost of treating patients with abortion complications and provider and abortion patient perspectives. Data were collected via one on one interviews and reviews of logbook data. A computerized literature review on abortion in the region covering the years 1980-1994 complemented the primary data collection. STUDY SETTING: Primary data collection was conducted in three countries (Zambia, Uganda and Malawi) at selected districts and tertiary care hospitals. SUBJECTS: Nurses and physicians providing care to women with complications of abortion in participating hospitals were interviewed as well as hospital administrators. Women receiving care in the selected hospitals during the data collection period also were interviewed. MAIN OUTCOME MEASURES: This analysis focused on cross-country comparisons of nurse and doctor attitudes and knowledge about abortion and family planning-related issues. INTERVENTIONS: At a 1994 CHRCS conference, Health Ministers from 12 Commonwealth countries reviewed and endorsed the results of this study. In a 1995 ECSA Director Joint Consultative Committee meeting, participants discussed the study findings and developed regional action plans which were subsequently endorsed by the Health Ministers' at their 1995 annual conference. RESULTS: Abortion complication patients in the three countries were on average 24-26 years old with two children. Patients experienced long hospital stays while receiving care for complications. The average monthly number of post-abortion patients was substantially higher in urban facilities as compared to mixed/rural ones. Opinions regarding factors that led to the abortion, access to abortion services and the legal provisions surrounding abortion were inconsistent between doctors and nurses. Patients were aware that complications of abortion were a major cause of maternal mortality. Overall, provider opinion about access to family planning (FP) information among abortion complication patients was also inconsistent between doctors and nurses but there was agreement on factors influencing use of FP. Opinions regarding access to FP services both before and after the abortion varied from easy to very difficult to obtain. CONCLUSIONS: These data represent the situation as it was in the region some years ago and the paper describes important implications of the findings for policy and programme development. This research provided some impetus for stakeholders in these countries to put safe abortion and management of abortion complications on their health agenda. Ensuring that research results will be shared with appropriate decision-makers is key to maximizing the extent to which research findings may affect policy and programme advocacy. PMID- 15125089 TI - Induced abortion among women attending antenatal clinics in Yaounde, Cameroon. AB - OBJECTIVES: Unsafe abortion is a public health concern because of its impact on maternal morbidity and mortality. The objective of this study was to document on induced abortion in Yaounde, Cameroon. DESIGN: Cross-sectional study. SETTING: Six antenatal clinics in Yaounde, Cameroon. METHODS: Women attending antenatal clinics between October and December 1998 were included in the study and interviewed. Nulliparous were women with no previous delivery and multiparous were defined as women who had at least one previous delivery. RESULTS: Out of the 1532 women, five hundred seventy-two were nulliparous and 960 were multiparous. Of the nulliparous women 17% reported a previous abortion ever; this proportion exceeded 35% in those over 24 years. For multiparous women, the proportion who reported an abortion (between the last birth and present pregnancy) was 22%. In multivariate analysis on the group of nulliparous women, older age, having used modern contraception and having spent more than two years in the city were significantly associated with induced abortion. In the multiparous group, older age, having a full time job and antenatal clinic were significantly associated with induced abortion. CONCLUSIONS: This study shows that induced abortion is a common practice in urban Cameroon. Because of restrictive laws, a substantial proportion of these abortions are likely to be unsafe, with the risk of associated complications. There is a need for expanded comprehensive sexual and reproductive health services. PMID- 15125090 TI - A stromal myoid cell line provokes thymic erythropoiesis between 16th to 20th weeks of intrauterine life. AB - BACKGROUND: The thymus provides an optimal cellular and humoral microenvironment for cell line committed differentiation of haematopoietic stem cells. The immigration process requires the secretion of at least one peptide called thymotaxine by cells of the reticulo-epithelial (RE) network of the thymic stromal cellular microenvironment. The thymic RE cells are functionally specialised based on their intrathymic location and this differentiation is modulated by various interaction signals of differentiating thymocytes and other non lymphatic haematopoietic stem cells. OBJECTIVES: To study the role of another cell line in fetal thymic haematopoietic proliferation and differentiation in different stages of development: the stromal myoid cells. DESIGN: Fifteen cases of fetal thymic specimens (4th to 8th weeks: five cases 16th to 20th weeks: five cases and 28th to 32nd weeks: five cases respectively) were studied. Tissue paraffin samples were stained immunohistochemically using (i) a monoclonal antibody recognising alpha-smooth muscle actin, a contractile microfilament expressed exclusively by smooth muscle cells, myofibroblasts and related cells, (ii) a monoclonal antibody glycophorin C recognising the erythropoietic cells. SETTING: Histology-Embryology Department of Democritus University of Thrace (Alexandroupolis) over ten year period (1991-2001). RESULTS: The number of alpha smooth muscle actin-positive cells significantly increased during the late second and third trimester of gestation. In the above period a relevant increase in the number of glycophorin C positive cells were observed. CONCLUSION: Our data suggest that a myoid cell line is involved in the formation of an appropriate microenvironment for homing and proliferation of erythropoietic cells. PMID- 15125091 TI - Cardiovascular risk factor profile of black Africans undergoing coronary angiography. AB - BACKGROUND: Coronary artery disease (CAD) is a growing epidemic on the African continent. It remains uncertain whether the risk factors identified as contributing to CAD in white populations contribute to a similar extent to CAD incidence in black populations. No data of the local population exists that is based on the coronary angiogram (CA). OBJECTIVES: To analyse the relationship of conventional cardiovascular risk factors with presence of CAD in black Africans. DESIGN: This was a dual-armed study, consisting of retrospective and prospective comparative arms. SUBJECTS: Black Africans who underwent coronary angiography. SETTING: Nairobi Hospital, Cathereterization laboratory. MAIN OUTCOME MEASURES: The conventional risk factors: age, male gender, hypertension, obesity, smoking, diabetes mellitus, dyslipidaemia, alcohol use and interventricular septum (IVS) hypertrophy, as a marker of LVH. RESULTS: One hundred and sixty nine patients fulfilled the inclusion criteria; 144 in the retrospective arm and 25 in the prospective. The larger retrospective arm showed that the group with CAD, compared to the normal group, was significantly older, with a higher mean age of 54.4 years compared to 49.8 years (P=0.005); had significantly more males, with a male to female ratio of 5.5:1 compared to 2.3:1 (P=0.045); had a very significantly larger proportion of diabetics (38.5% compared to 12%, P=0.0002), and also had a significantly larger proportion of patients with dyslipidaemia (67.3% compared to 35.9%, P=0.0003). The percentage of hypertensives was high in both groups, with (65.4%) in the CAD group and 62% in the Normal group being hypertensive (P=0.68). The percentage of smokers was small in both groups, being 15.4% and 13% respectively. Smoking, increased BMI, alcohol use, and increased IVS were each found to be distributed equally in both groups. In addition, the Waist hip ratio (WHR) and waist circumference (WC) did not differ significantly between the two groups studied. CONCLUSIONS: The risk factors found to be most strongly associated with presence of angiographically-detected CAD in the population studied were diabetes mellitus, dyslipidaemia, age and male gender. There was a high prevalence of hypertension, with equal distribution in both groups under study; hence this risk factor was not discriminatory for CAD. There was a low prevalence of cigarette smoking in this particular study; it was not predictive of presence of CAD. PMID- 15125092 TI - Managing scalp defects in sub-Saharan Africa. AB - OBJECTIVES: To determine the common aetiological factors of scalp defects, and outcome of management. DESIGN: A two year prospective study. SETTING: Usmanu Danfodiyo University Teaching Hospital, Sokoto, Nigeria. SUBJECTS: All consecutive patients with scalp defect from January 2001 to December 2002. MAIN OUTCOME MEASURES: Size of defect, associated bone loss, osteomyelitis, type of surgery, duration of hospital stay and complications of surgery. INTERVENTIONS: Patients went through a simple management protocol involving history, clinical examination, relevant tests and appropriate treatment, including surgery. RESULTS: A total of 27 patients were studied out of which 15 were males and 12 females, giving a male to female ratio of 1.25:1. The age range was seven months 42 years (mean = 13.9 years). Road Traffic Accidents (RTAs) was the commonest cause of scalp defects (81.5%). The temporo-parietal area was involved in over 50% of patients. Chronicity and osteomyelitis were common complications of the defects. Over 50% of the patients had local flap reconstruction. CONCLUSIONS: Management of scalp defects remains a major challenge in our environment. The importance of continuing education of colleagues and other health workers in peripheral health units on the importance of proper initial wound debridement and early referral cannot be overemphasised. PMID- 15125093 TI - Predictors for compliance with community directed ivermectin treatment in Bushenyi district of Uganda: qualitative results. AB - OBJECTIVE: To understand and elicit the factors influencing compliance with mass treatment with ivermectin for onchocerciasis control with a view of suggesting remedial measures. DESIGN: Qualitative methods using focus group discussion and individual key informant interviews. SETTING: Bushenyi district, Uganda. PARTICIPANTS: Fifty key informants who were local council chairpersons and community directed drug distributors (CDDs) for ivermectin. Five focus group discussions with community members. MAIN OUTCOME MEASURES: Explored knowledge about onchocerciasis and its treatment, consequences, benefits and perceived dangers of treatment with ivermectin. In addition the FGDs elicited information regarding social influence towards ivermectin treatment and who the source of social influences are. Perceived barriers and supports towards ivermectin treatment were also explored. The key informant interviews mainly elicited the problems they face in ivermectin distribution and their suggestions for improvement. RESULTS: Factors that could possibly influence mass treatment with ivermectin that were elicited include consequences of treatment, programme organization, charging for ivermectin distribution and programme/community support to the ivermectin drug distributors. CONCLUSIONS: These results are helpful in redirecting community education and in helping to design further quantitative research. PMID- 15125094 TI - Leishmania major-Phlebotomus duboscqi interactions: inhibition of anti-LPG antibodies and characterisation of two proteins with shared epitopes. AB - OBJECTIVES: To assess the effect of monoclonal antibodies (MABS) raised against L. major derived LPG on L. major development in vitro and in its natural vector P. duboscqi. Also determine whether LPG molecule and the sand fly the gut lysates have shared epitopes. DESIGN: A laboratory based study. SETTING: Colony bred P. duboscqi sand flies and all other experiments were done under laboratory conditions. METHODS: Laboratory reared sand flies were allowed to feed beneath a blood filled membrane feeder containing 1 x 10(6) amastigotes in 20 microl mixed with 0.5 ml of defibrinated rabbit blood with a 1:100 dilution of anti-LPG MABS. Control blood contained a similar number of amastigotes but no MABS. At least five female previously fed sand flies were later dissected on days two, four, and six post-feeding and examined for promastigote forms and parasite loads in the sand fly mid gut. In vitro, the same number of amastigotes in 100 microl complete Schneider's Drosophila medium was mixed in a 96 well plate with either 100 microl of 1:100 anti-LPG MABS, 1:1000 anti LPG MABS or undiluted sera from L. major infected mice. The control well contained a similar number of amastigotes but no antibodies added. Following an overnight incubation in a CO2 incubator at 37 degrees C and growth at 26 degrees C, parasites were assessed at 3, 6 and 24 hour intervals for changes in their developmental forms. RESULTS: 1:100 dilution of anti-LPG MABS when mixed with amastigotes were effective in reducing L. major development at the early log phase or procyclic stage both in vitro and within the sand fly (p<0.05). The control cultures or sand flies that fed on amastigotes alone and no MABS supported full parasite development up to the metacyclic stage. Results also showed that flies, which had fed on MABS, showed low parasitemia levels of 2+, compared to a high density of 4+ for their controls (p<0.5). CONCLUSIONS: These findings showed that anti-LPG MABS were effective in reducing sand fly infections. This study also showed that P. duboscqi gut lysates and proteins present in L. major-derived LPG share two common proteins of molecular weights 105 kDa and 106 kDa. Further analysis of these individual proteins from the gut should be studied with a view of determining their vaccine potential. PMID- 15125095 TI - Diagnostic value of plain abdominal radiographs in acute appendicitis in children. AB - OBJECTIVE: The plain film of the abdomen (PAX) is still utilised in the diagnosis of acute appendicitis (Aap). Aim of this study was to evaluate the value of PAX in the diagnosis of Aap in children, since it continues to be a controversial subject. DESIGN: A retrospective study. SETTING: Department of Paediatric Surgery, Gazi University Medical School. PATIENTS: The histopathological diagnoses and PAX of 213 children (mean age 9-6 years) operated for acute right lower quadrant pain were evaluated retrospectively. There were 181 patients with Aap and 32 with normal appendix. METHODS: The rate of detection of overall thirteen roentgenographic criteria defined in various reports in medical literature as indicating appendicitis were noted and compared with histopathological diagnoses. The PAX which revealed one or more of these signs in combination was considered to be positive for Aap. Data was analysed statistically and sensitivity, specificity and positive and negative predictive values of PAX in the diagnosis of Aap were determined. RESULTS: Roentgenographic sign(s) were detected in 170 (79%) cases, 18 without Aap. The most frequent were lumbar scoliosis with left-sided convexity (50%) and small intestinal air-fluid levels (32%). In 39 cases, 25 with Aap, no sign(s) was detected. In all of the perforated cases, at least one of the determined criteria was seen. The percentage of roentgenographic signs found in cases with and without appendicitis was different significantly (p<0.01). The negative predictive value of PAX in the diagnosis of acute appendicitis was 32.5% and positive predictive value was 89%. CONCLUSION: PAX could he helpful in diagnosis in children with suspected acute appendicitis. PMID- 15125096 TI - Non-surgical management of a chronic periapical lesion associated with traumatised maxillary central incisors: case report. AB - A case is reported of a chronic periapical lesion involving maxillary central incisors with a history of traumatic injury eight years previously and subsequent development of a painful swelling that occasionally caused partial blockage of the nasal cavities. Retrograde surgery for removal of the suspected cystic lesion was scheduled. As a temporary measure before surgery, the necrotic pulp exudate was removed through standard endodontic access cavities prepared on the palatal surfaces of the crowns of the two central incisors, and a calcium hydroxide paste dressing material was placed inside the root canals up to the apices. The calcium hydroxide dressing was changed every four weeks and the case was followed up until healing of the periapical lesion occurred. The surgical procedure was postponed and finally cancelled when complete healing became evident in recall radiographs. PMID- 15125097 TI - Why malaria control is failing in Africa and the suggested way forward. PMID- 15125098 TI - Is telehealth the right tool for remote communities? Improving health status in rural Australia. AB - The health status of people in rural and remote areas, particularly of Indigenous Australians, has been widely reported to be poorer than their urban counterparts. Health care in rural and remote Australia has relied on the work of nurses, often in the absence of medical or allied health personnel other than via telecommunication. Over the last 5 years telehealth has been heavily promoted as an innovative and effective way of improving the health status of people in rural and remote areas by providing improved access to specialist care. Through national infrastructure development and funding for a vast array of pilot projects, telehealth provision has been promoted as progressive, effective, and modern. This Editorial reviews the literature and highlights a need for additional research around telehealth-mediated patient practitioner relationships. PMID- 15125099 TI - Nursing in the aged care arena: perceptions of the experienced gerontic nurse. AB - This qualitative research project explored the complexities associated with providing competent care to older Australians, especially within the context of the current health care environment. The study used the perceptions of experienced Registered Nurses in a residential aged care facility to highlight the ways in which the quality of care in these facilities is both driven and maintained by the Registered Nurse. It is argued that Registered Nurses are the linchpins who act as providers and coordinators of care within residential aged care services. The paper focuses on the potential for the development of the role of the gerontological nurse in residential aged care, and provides future direction for practice. A feature of the study was the way in which the five participants presented an overwhelming respect and value for the older person. PMID- 15125100 TI - Influences on positive family involvement in aged care: an ethnographic view. AB - This paper examines the experiences of family members who have previously been primary carers of residents of residential aged care services to identify factors, which have encouraged them to remain involved in a caring role. The context for this study was a residential aged care facility in Western Australia that was selected on the basis of its reputation of being highly sought after by relatives requiring formal care for older family members and also having a high level of ongoing involvement of relatives. Findings from the study provide evidence of the importance of staff developing relationships with relatives, supporting relatives through the change process and developing care partnerships where relatives could define new caring roles in providing social and emotional support for residents. This study is part of a larger study investigating relational quality in aged care environment. PMID- 15125101 TI - Under the circumstances: the experiences of younger people living in residential aged care facilities. AB - This small-scale, exploratory, descriptive study recorded the experiences of people under the age of 65 years living in residential aged care facilities. Individuals under the age of 65 are significantly younger than the majority of residents in residential aged care facilities. The findings of this study indicated that under the circumstances participants often perceived they had no alternative living arrangements available to them. Being a younger person in an aged care facility effected emotional wellbeing, relationships with others and activities, such as recreation. These findings provide information that increases knowledge, understanding and empathy. In addition they can inform residential care policy and influence nursing care in residential aged care facilities. The findings also provide a foundation for further research. PMID- 15125102 TI - Community health nursing in Australia: a critical literature review and implications for professional development. AB - BACKGROUND: Increasing emphasis on community-based mandates an examination of the community health nurse (CHN). AIM: A critical literature review of the CHN role internationally, with an emphasis on Australia, was undertaken in order to understand historical precedents and inform policy and strategic directions for the CHN. METHOD: A search of the CINAHL, EMBASE, and COCHRANE electronic data bases from 1982 to June 2002 using community' and 'nursing' as key words and hand searching of books and government reports was undertaken. FINDINGS: This search strategy revealed a lack of literature compared with other areas of nursing. Key themes emerging from this review are: (1) an absence of clear role definitions and lack of clarity of roles; (2) variability in educational requirements for CHNs; (3) diminished CHN power in policy decision making; (3) conflicting role expectations between different facets of the health care system; (4) underutilisation and untapped potential of the role of the CHN in the contemporary health care system; (5) the emerging influence of specialist nurses in community based-care; (6) uptake of traditional nursing roles by non-nurses and (7) an absence of a cohesive model of professional development of CHN that is able to articulate with contemporary social, political and economic trends in health care delivery. CONCLUSIONS: Community health nursing in Australia has a low professional profile when compared to other nursing specialties. An emerging issue, gleaned from the literature review is the tension and debate between specialist and generalist services. It is apparent from this review that CHN have to more actively participate in research and peer reviewed debate in order to have their voice heard and promote their unique and valuable contribution to the nursing profession and the health care system. PMID- 15125103 TI - Changing focus of practice for community health nurses: advancing the practice role. AB - Many parents lack support in their parenting role that was once provided through extended families and community structures. Thus, some new parents experience high levels of stress and low self-esteem associated with the challenges of parenting. The lack of support also results in family discord and breakdown with the family environment having the potential to adversely impact children's mental and physical wellbeing and development. The Community Mothers Program (CMP) was initially developed in England and offers support to families during the first year of parenting. The program aims to provide parents with the support once experienced from within the extended family. It also aims to enrich community development by building the capacity of community members living in local communities to support parents. This paper describes the impact of the CMP when implemented into Western Australian as well as the changes to the professional practice role of community child health nurses involved in the program. The Community Mothers Program has proved to be very successful. The success is attributed to the partnership model established between community members, parents, and child health nurses. PMID- 15125104 TI - Nurses' knowledge and learning experiences in relation to the effects of domestic abuse on the mental health of children and adolescents. AB - Nurse researchers are yet to direct substantial attention towards addressing and understanding nurses' experiences of learning about and caring for child and adolescent victims of domestic abuse. This lack of recognition has resulted in the marginalisation of this issue. This paper seeks to explore nurses' present understanding and experiences of learning about the effects of domestic abuse on the mental health of children and adolescents. The research utilised an interpretive case study approach within a naturalistic paradigm. Snowball sampling of nursing staff within two mental health units was used. Data were collected using semi-structured in-depth interviews with each participant. Using a constant comparative method and hermeneutic dialectical process both the commonalities and differences regarding participants' multiple realities were identified. Three major categories emerged within the data. These were education, resources and nurses' role. It was apparent that nurses' knowledge and education about the effects of domestic abuse on the mental health of children and adolescents negatively impacted on nurses' ability to provide appropriate care. The research has clear implications for mental health nursing education, practice and future research. PMID- 15125105 TI - "Don't forget your cotton underwear girls": femininity and Australian Army nurses in the Vietnam War (1967-1971). AB - War and nursing are unequivocally linked and the impact that this type of nursing has had upon the nursing profession in indisputable. However, a review of the Australian scholarly nursing literature revealed that the contribution and experiences of Australian nurses in the Vietnam War has not been widely published. The direct involvement of Australian women in the Vietnam War was limited and, as female nurses, they were unquestionably a minority. This paper describes the gender issues that confronted the nurses who worked within the confines of war. PMID- 15125107 TI - The sonata form of musical composition as a framework for thesis writing. AB - In response to Watson's challenge to view the usual in new ways, this article introduces an innovation in writing Master's research and suggests that other structures may offer new and different frameworks for reporting nursing research. This is exemplified by reference to an example of nursing research which adopted the sonata-form of musical composition as the framework for presentation of the thesis. PMID- 15125106 TI - Collecting qualitative data: Part II. Group discussion as a method: experience, rationale and limitations [corrected]. AB - This article discusses the experience, rationale for and limitations of the group discussion as a method for collecting qualitative data. The researcher defines the group discussion as a focused group depth discussion' premised on a review of the focus group literature. A number of strengths are proposed for and the limitations are countered in the use of the group discussion as a method for collecting qualitative data. PMID- 15125108 TI - Acting on rational drug use in Uganda. The interface between research and policy. PMID- 15125109 TI - Impact of decentralization on health services in Uganda: a look at facility utilization, prescribing and availability of essential drugs. AB - INTRODUCTION: Uganda began implementation of a structural adjustment programme (SAP) in July 1994 in order to improve social services. The decentralization of health services administration to district level was intended to improve the quality of health services and pharmaceutical supplies in the hospitals, with resultant increase in the level of utilization of health facilities. OBJECTIVE: This study evaluated the impact of the decentralization policy on health facility utilization; availability of essential drugs, and prescribing patterns for acute respiratory infections (ARI), diarrhoea, and malaria in two district hospitals in Uganda. DESIGN: Mixed method evaluation design, involving both quantitative and qualitative methods. Time series analyses of data from utilization, pharmacy stock, and prescription records before and after the policy change. Key informant interviews and focus group discussions to obtain information on perceptions and attitude of stakeholders on the process of the policy implementation. STUDY SETTING AND POPULATION: The study was conducted in two district hospitals in northern Uganda. A total of seven years of utilization and pharmacy stock data including 5040 patient records from the hospitals were analysed retrospectively. In-depth interviews were conducted among 11 politicians from each district; 100 open-ended questionnaires were administered to patients in each hospital; 86 health care workers were interviewed using semi-structured questionnaires; and focus group discussions were conducted with 23 health care providers. MAIN OUTCOME MEASURES: Facility utilization was evaluated by average monthly attendance in the outpatient department and paediatric ward admissions. Availability was assessed as average number of drugs per month. Prescribing indicator outcomes included: for malaria, percent chloroquine tablets and percent chloroquine injection; for ARI, percent receiving antibiotics or injections; for diarrhoea, use of oral rehydration salts (ORS), antidiarrhoeal mixtures, and antibiotics. The average number of drugs prescribed assessed polypharmacy. RESULTS: There was a general increase in patient attendance in both hospitals, although the initial increase later declined in Apac. Drug availability was erratic and not always adequate. The situation was better in Lira where funding for drug procurement was more accessible. Prescribing patterns varied, with improvement in some indicators, while others showed no change or even worsened. CONCLUSIONS: The decentralization policy led to increased utilization of health facilities. The perception was that the policy was good because it "empowered the community in terms of creating a sense of responsibility in the stakeholders, and a sense of ownership that facilitated sustainability" of public institutions. In spite of the views expressed by the stakeholders, the policy failed to improve drug shortages, inefficient utilization of resources, and low morale among hospital staff. Staff should be re-trained and better remunerated in order to cope with the implementation of the policy. Local politicians should clearly understand their roles and responsibility under the new policy. Efficient utilization of funds at all levels of the district administrative structures should be ensured. PMID- 15125110 TI - Attitudes and perceptions of stakeholders on decentralization of health services in Uganda: the case of Lira and Apac districts. AB - BACKGROUND: In Uganda, the decentralization of administrative functions, management, and responsibility for health care to districts, which began in 1994, resulted in fundamental changes in health care delivery. Since the introduction of the policy in Uganda, little information has been available on stakeholders' perceptions about the benefits of the policy and how decentralization affected health care delivery. OBJECTIVES: To identify the perceptions and beliefs of key stakeholders on the impact and process of decentralization and on the operations of health services in two districts in Uganda, and to report their suggestions to improve future implementation of similar policies. DESIGN: We used qualitative research methods that included focus group discussions with 90 stakeholders from both study districts. SETTING: The sample population comprised of 12 health workers from the two hospitals, 11 district health administrators, and 67 Local Council Leaders. MAIN OUTCOME MEASURES: Perceptions and concerns of stakeholders on the impact of decentralization on district health services. RESULTS: There was a general consensus that decentralization empowered local administrative and political decision-making. Among stakeholders, the policy was perceived to have created a sense of ownership and responsibility. Major problems that were said to be associated with decentralization included political harassment of civil servants, increased nepotism, inadequate financial resources, and mismanagement of resources. CONCLUSIONS: This study elicited perceptions about critical factors upon which successful implementation of the decentralization policy depended. These included: appreciation of the role of all stakeholders by district politicians; adequate availability and efficient utilization of resources; reasonably developed infrastructure prior to the policy change; appropriate sensitisation and training of those implementing policies; and the good will and active involvement of the local community. In the absence of these factors, implementation of decentralization of services to districts may not immediately make economic and administrative sense. PMID- 15125111 TI - A comparison of prescribing practices between public and private sector physicians in Uganda. AB - INTRODUCTION: Previous studies in the public sector in Uganda have demonstrated major prescribing problems due to polypharmacy and irrational use of antibiotics and injections. Little is known about prescribing in the private sector although there is little government regulation influencing practice in this sector. The introduction of policies such as the Uganda National Standard Treatment Guidelines (UNSTG) was expected to improve prescribing practices in the public and private sectors. This paper measures appropriateness of prescribing practices in the public and private sectors in Uganda for the treatment of Acute Respiratory Infections (ARI) and malaria in adult patients. DESIGN AND SAMPLING: We combined a prospective survey of treatment for simulated patients presenting with symptoms of malaria and ARI in 119 randomly selected private clinics and a retrospective survey of 600 prescription records for malaria and ARI (300 for each condition) randomly selected from 10 public health units in the Kampala, Masaka, and Jinja urban areas. OUTCOME MEASURES: Percentage of drug appropriately prescribed in each condition, % injection prescription, percentage antibiotic prescription, average number of drugs per case, average standardized cost per prescription, distribution of types of drugs prescribed. RESULTS: The overall appropriateness of prescribing for ARI and malaria was poor in both public and private sectors. Treatment of malaria was significantly less appropriate in the public sector compared to the private sector (14% vs. 27%, p = 0.002), with injectable chloroquine much more commonly prescribed. Prescribing of antibiotics for ARI was nearly universal in both sectors, with some prescriptions containing up to three antibiotics; newer, more expensive antibiotics were more commonly prescribed in the private sector. Polypharmacy and unnecessary prescribing of vitamins were common in both conditions and both sectors. CONCLUSIONS: Prescribing for adult malaria and ARI by both private and public practitioners did not conform to the UNSTGs. Although practitioners were largely the same in both sectors, prescribing practices often differed dramatically. The extent of inappropriate prescribing in both sectors calls for in-depth investigation of the system factors and motivations that underlie problem practices, and the development of interventions that target these causative factors. PMID- 15125112 TI - Impact of an educational intervention to improve prescribing by private physicians in Uganda. AB - INTRODUCTION: Private physicians in urban Uganda treat a large percentage of common adult illnesses. Improving their prescribing would not only encourage more rational drug use, but also reduce costs to patients. Interventions to improve drug use are generally more successful when face-to-face educational methods are included. OBJECTIVES: To determine the effectiveness of a face-to-face educational intervention on the treatment of acute respiratory infections (ARI), malaria, and non-dysenteric diarrhoea by private physicians in three urban areas of Uganda. METHODS: The study used an intervention with comparison group design to evaluate the impact of the educational intervention. A total of 108 private physicians was divided into intervention (n = 30) and control (n = 78) groups. Surrogate patients, trained to simulate presenting symptoms and signs of the target conditions, were used to collect data on the medical practices and prescribing behaviours of the physicians. Intervention physicians were invited to a one-day interactive educational seminar facilitated by local opinion leaders that covered principles of rational drug use and the National Standard Treatment Guidelines for treating the target conditions. Physicians were also provided with data about baseline practices. RESULTS: Baseline data indicated high rates of inappropriate treatment practices by both intervention and control groups. There was nearly universal antibiotic use for ARI (over 90%), high rates of injections recommended for malaria (over 30%), and high rates of polypharmacy (over thee drugs per patient). After the intervention, some significant improvements in key practices were observed in the intervention group. Compared to control physicians, antibiotic prescribing for ARI decreased by 23% in the intervention group, use of combination products for malaria declined by 28%, there were trends towards better adherence to guidelines for ARI and malaria, and marginal decreases in drug costs. However, the overall impacts of the intervention were limited, especially on quality indicators concerning history taking, adequate examination, and advice to patients. CONCLUSION: The face-to-face educational intervention resulted in some small improvements in key prescribing practices of private physicians. However, an intervention that involved repeated contacts with prescribers and which addressed economic considerations would be needed to obtain larger improvements. Private physicians need to be sensitized to and encouraged to use the National Standard Treatment Guidelines, and attempts to improve their prescribing should be supported by community education. PMID- 15125113 TI - Impact of a face-to-face educational intervention on improving the management of acute respiratory infections in private pharmacies and drug shops in Uganda. AB - BACKGROUND: In Uganda, although private retail pharmacies and drug shops are the most common sources of drugs for the majority of the population, the quality of care received from these outlets has been reported as suboptimal. It is believed that lack of adequate knowledge is an important underlying factor to such practices. OBJECTIVES: In this study, we investigated the impact of a face-to face educational intervention on counter attendants' dispensing behaviour for mild and severe acute respiratory infections (ARI) in children at private pharmacies and drug shops. STUDY DESIGN AND SETTING: We used a quasi-experimental research design with comparison groups to analyse counter attendants' management of ARI before and after an intervention. The study was conducted in Kampala District, and used a sample of 191 registered drug outlets (27 pharmacies and 164 drug shops), stratified into two groups: 1) An experimental group that received training; 2) A control group unexposed to training. Data on the practices in the drug outlets from both groups were collected at two time-points: seven months before the intervention, and at one-month after the intervention. OUTCOME MEASURES: Two main outcome measures were considered: a) Assessment of the child's condition. b) The dispensing practices of counter attendants. RESULTS: The study found that despite the training, the assessment of the child's condition remained inadequate in both groups, where the child's age was the only question asked in more than 90% of cases. High levels of inappropriate dispensing practices for both mild and severe ARI were still persistent in both groups after the intervention. Antibiotic prescribing for both conditions was very common, and barely any advice or instruction was given with dispensed drugs. Client demand for particular drugs, competition among drug outlets, and inability of most clients to afford the recommended treatments were the main reported barriers that emerged from the focus group discussions with the counter attendants. CONCLUSIONS: The evaluation of the practices one month after the face-to-face educational intervention showed that the management of ARI did not improve in the drug outlets. While study design issues may have contributed to such findings, there are many other factors not related to knowledge and education that may indirectly hamper the promotion of appropriate dispensing in the private pharmacies and drug shops in Uganda. It is possible that a combination of interventions may contribute to improved management of ARI by counter attendants in the private drug shops and pharmacies in Uganda. PMID- 15125114 TI - Management of acute respiratory infections in drug shops and private pharmacies in Uganda: a study of counter attendants' knowledge and reported behaviour. AB - BACKGROUND: In Uganda, private pharmacies and drug shops are important sources of drugs for the majority of the population. In addition to selling drugs, these outlets often serve as primary sources of information about illness and drug therapy. However, the appropriateness of dispensing by staff in these drug outlets has been found to be suboptimal. Yet there has been no study documenting the determinants that underlie the dispensing pattern at these outlets. This study evaluated counter attendants' training background, their knowledge of acute respiratory infections (ARI), and their reported behaviour in the management of ARI in order to identify factors associated with dispensing behaviours. STUDY DESIGN AND SETTING: This descriptive study was conducted in Kampala District, Uganda and used a cross-sectional survey research design to analyse the practice of 197 drug outlets including 28 pharmacies and 169 drug stores. MAIN OUTCOME MEASURES: Counter attendants' training background; their knowledge of the causes, signs and symptoms of ARI; their perception of the dangers of ARI; and the drugs and advice offered for the management of mild and severe ARI. RESULTS: Majority of the counter attendants had medical or pharmacy training. Most of them were nurses. The attendants' workload was generally low. High levels of self medication among clients were reported at the outlets. Staff at a management level had low knowledge of the aetiology, signs and symptoms, and dangers of ARI. Prescribing an antibiotic was found to be the usual practice for managing ARI cases. Counter attendants at those outlets seldom gave advice or referrals for ARI cases. In particular, patient demand was a main barrier to appropriate dispensing. Strategies suggested by attendants for improving rational dispensing were to educate the public, provide training for the attendants, and re-enforce government supervision of drug outlets. CONCLUSIONS: This study identified several self-reported inappropriate behaviours in the outlets. The formal training background and workload of counter attendants at drug shops and pharmacies were not found to be important contributors to irrational prescribing. The results of this study suggest that a combination of regulatory and educational interventions may yield to improvement in counter attendants' practices in private drug outlets. PMID- 15125115 TI - Impacting drug policy in Uganda. PMID- 15125116 TI - Inflammatory serum markers in patients with multiple trauma. Can they predict outcome? PMID- 15125117 TI - Posterior dislocation of the shoulder. PMID- 15125118 TI - Radiation protection issues with the use of mini C-arm image intensifiers in surgery in the upper limb. Optimisation of practice and the impact of new regulations. PMID- 15125119 TI - The use of structural proximal femoral allografts in complex revision hip arthroplasty. AB - Between April 1992 and November 1998 we used 34 massive proximal femoral allografts for femoral reconstruction at revision hip arthroplasty. Seven patients have died and two have been lost to follow-up. There were thus 25 grafts in 24 patients for review. The mean follow-up was 53 months (16 to 101). By the time of the review two patients had undergone a further revision for failure of the allograft. Another had required secondary plating and grafting at the graft host junction for symptomatic nonunion. One had recurrence of deep sepsis and was being managed conservatively. Trochanteric union was considered to have occurred radiologically in 16 of the 25 grafts and union at the host-graft junction in 20. Resorption of the allograft was significant in only two hips. We recommend this technique in cases in which femoral bone loss has been catastrophic. PMID- 15125120 TI - A comparison of polyethylene wear rates between cemented and cementless cups. A prospective, randomised trial. AB - We selected randomly a consecutive series of 162 patients requiring hip replacement to receive either a cementless, hemispherical, modular, titanium acetabular cup or a cemented, all-polyethylene cup. These replacements were performed by two surgeons in four general hospitals. The same surgical technique was used and a 26 mm metal-head femoral component was used in every case. After exclusions, 115 hips were studied for differences in rates of wear and osteolysis. The mean clinical follow-up was eight years and the mean radiological follow-up, 6.5 years. The cementless cups wore at a mean rate of 0.15 mm per year and the cemented cups at 0.07 mm per year. This difference was significant (p < 0.0001). Our findings in this mid-term study suggest that cementless cups wear more than cemented cups. PMID- 15125121 TI - The effects of calcitonin on acute bone loss after pertrochanteric fractures. A prospective, randomised trial. AB - We investigated the effect of calcitonin in the prevention of acute bone loss after a pertrochanteric fracture and its ability to reduce the incidence of further fractures in the same patient. Fifty women aged between 70 and 80 years who had a pertrochanteric fracture of the hip were randomly allocated to group A (200 IU of nasal salmon calcitonin daily for three months) or group B (placebo). Patients in group A showed a significantly higher level of total alkaline phosphatase and osteocalcin on the 15th day after injury and a significantly higher level of bone alkaline phosphatase on the 90th day after surgery. These patients also had significantly lower levels of urinary C-telopeptide (CrossLaps) on the 15th, 45th and 90th days after injury and lower levels of urinary hydroxyproline on the 15th and 45th days after injury. Patients in group A had significantly higher bone mineral density at all recorded sites except the greater trochanter at three months and one year after operation. After a four year period of clinical observation, five patients (24%) in group B sustained a new fracture, in four of whom (20%) it was of the contralateral hip. Our findings show that calcitonin reduces acute bone loss in patients with pertrochanteric fractures and may prevent the occurrence of new fractures of the contralateral hip in the elderly. PMID- 15125122 TI - Curved intertrochanteric varus osteotomy for osteonecrosis of the femoral head. AB - We reviewed the outcome of curved intertrochanteric varus osteotomy in the treatment of osteonecrosis of the femoral head in 20 hips. A mean varus angulation of 31 degrees was obtained by the osteotomy. The ratio of intact area on the weight-bearing portion increased from 19% to 61%. The mean elevation and lateral displacement of the greater trochanter were 1.2 cm and 0.5 cm, respectively. These changes in the position of the greater trochanter were very small when compared with those after conventional varus wedge osteotomy. Nonunion or delayed union was not observed. Quantitative analyses showed aggressive bone remodelling in the medial intertrochanteric region. Eighteen hips survived without collapse after a mean follow-up of 48 months. We conclude that curved varus osteotomy can be used to preserve the hip joint in patients with osteonecrosis of the femoral head. PMID- 15125123 TI - The accuracy of image-guided knee replacement based on computed tomography. AB - Our study evaluated the accuracy of an image-guided total knee replacement system based on CT with regard to preparation of the femoral and tibial bone using nine limbs from five cadavers. The accuracy was assessed by direct measurement using an extramedullary alignment rod without radiographs. The mean angular errors of the femur and tibia, which represent angular gaps from the real mechanical axis in the coronal plane, were 0.3 degrees and 1.1 degrees, respectively. The CT based system, provided almost perfect alignment of the femoral component with less than 1 degrees of error and excellent alignment with less than 3 degrees of error for the tibial component. Our results suggest that standardisation of knee replacement by the use of this system will lead to improved long-term survival of total knee arthroplasty. PMID- 15125124 TI - Computer-assisted knee arthroplasty versus a conventional jig-based technique. A randomised, prospective trial. AB - We have compared a new technique of computer-assisted knee arthroplasty with the current conventional jig-based technique in 70 patients randomly allocated to receive either of the methods. Post-operative CT was performed according to the Perth CT Knee Arthroplasty protocol and pre- and post-operative Maquet views of the limb were taken. Intra-operative and peri-operative morbidity data were collected and blood loss measured. Post-operative CT showed a significant improvement in the alignment of the components using computer-assisted surgery in regard to femoral varus/valgus (p = 0.032), femoral rotation (p = 0.001), tibial varus/valgus (p = 0.047) tibial posterior slope (p = 0.0001), tibial rotation (p = 0.011) and femorotibial mismatch (p = 0.037). Standing alignment was also improved (p = 0.004) and blood loss was less (p = 0.0001). Computer-assisted surgery took longer with a mean increase of 13 minutes (p = 0.0001). PMID- 15125125 TI - Pattern of diabetic neuropathic arthropathy associated with the peripheral bone mineral density. AB - The relationship between the bone mineral density (BMD) and Charcot arthropathy is unclear. Prospectively, 55 consecutive diabetic patients presenting with a Charcot arthropathy of the foot or ankle were classified as having a fracture, dislocation, or a combination fracture-dislocation pattern of initial destruction. In these groups we used dual-energy x-ray absorptiometry to compare the peripheral bone of the affected and unaffected limbs. The clinical data relating to diabetes and related major comorbidities and the site of the arthropathy (ankle, hindfoot, midfoot, forefoot) were also compared. There were 23 patients with a fracture pattern, 23 with a dislocation pattern, and nine with a combination. The age-adjusted odds ratio for developing a Charcot joint with a fracture pattern as opposed to a dislocation pattern in patients with osteopenia was 9.5 (95% confidence interval 2.4 to 37.4; p = 0.0014). Groups also differed as to the site of the arthropathy. Fracture patterns predominated at the ankle and forefoot whereas dislocations did so in the midfoot. Diabetic Charcot arthropathy of the foot and ankle differs according to the pattern of the initial destruction. The fracture pattern is associated with peripheral deficiency of BMD. The dislocation pattern is associated with a normal BMD. PMID- 15125126 TI - Weakness of extensor hallucis longus after removal of non-vascularised fibula as an autograft. AB - The upper three-quarters of the fibula is commonly used as a non-vascularised autograft. Subsequent to this isolated weakness of extensor hallucis longus may occur. We have studied 26 patients in whom the upper and middle thirds of the fibula had been harvested as a graft through Henry's posterolateral approach. Isolated weakness of extensor hallucis longus was found after operation in ten patients but not in the remainder. EMG and nerve-conduction studies confirmed injury of the nerve to extensor hallucis longus in those with weakness. We dissected 40 cadaver limbs and found that those in which the nerve to extensor hallucis longus ran close to the fibular periosteum were at risk. The injury is mostly incomplete and recovery occurs within four to six months. PMID- 15125127 TI - Grammont inverted total shoulder arthroplasty in the treatment of glenohumeral osteoarthritis with massive rupture of the cuff. Results of a multicentre study of 80 shoulders. AB - We reviewed 80 shoulders (77 patients) at a mean follow-up of 44 months after insertion of a Grammont inverted shoulder prosthesis. Three implants had failed and had been revised. The mean Constant score had increased from 22.6 points pre operatively to 65.6 points at review. In 96% of these shoulders there was no or only minimal pain. The mean active forward elevation increased from 73 degrees to 138 degrees. The integrity of teres minor is essential for the recovery of external rotation and significantly influenced the Constant score. Five cases of aseptic loosening of the glenoid and seven of dissociation of the glenoid component were noted. This study confirms the promising early results obtained with the inverted prosthesis in the treatment of a cuff-tear arthropathy. It should be considered in the treatment of osteoarthritis with a massive tear of the cuff but should be reserved for elderly patients. PMID- 15125128 TI - Heterotopic ossification of the elbow in patients with burns. Results after early excision. AB - Heterotopic ossification which may develop around the elbow in patients with burns may lead to severe functional impairment. We describe the outcome of early excision of such heterotopic ossification in 28 patients (35 elbows), undertaken as soon as the patient's general and local condition allowed. The mean age at operation was 42 years. The mean area of burnt body surface was 49%. The mean pre operative range of movement was 22 degrees in flexion/extension and 94 degrees in pronation/supination. The mean time between the burn and operation was 12 months with the median being 9.5. The mean follow-up period was for 21 months. At the last review, the mean range of movement was 123 degrees in flexion/extension and 160 degrees in pronation/supination. Clinical evidence of recurrence was seen in four patients, occurring within the first two months after operation. Nevertheless, three of these elbows gained 60 degrees or more in flexion/extension and in pronation/supination. Based on this experience, we recommend early surgical treatment of heterotopic ossification of the elbow in patients with severe burns. PMID- 15125129 TI - Transient claw hand owing to a bee sting. A report of two cases. AB - We describe two patients with claw hand as a result of a bee sting. It is likely that this was caused by the apamin in the sting which has an effect on the upper limb, at the spinal cord and on the peripheral nerves. It is important to recognise that the claw hand is not owing to compartment syndrome. Both patients were treated conservatively with full resolution within 48 hours, without any lasting effects. PMID- 15125130 TI - Complex fracture-dislocation of the proximal interphalangeal joint of the hand. Results of a modified pins and rubbers traction system. AB - We reviewed 13 patients with a complex fracture-dislocation of the proximal interphalangeal joint of a finger and one patient with a complex fracture dislocation of the interphalangeal joint of thumb. We had treated these injuries using a pins and rubbers traction system which had been modified to avoid friction of the pins against the bone during mobilisation of the joint in order to minimise the risk of osteolysis. A Michigan hand outcome questionnaire was used for subjective assessment. The active range of movement (AROM) of the proximal and distal interphalangeal joints and the grip strength were used for objective assessment. The mean follow-up was 34 months (12 to 49). The mean normalised Michigan hand outcome score was 84. The mean AROM of the proximal interphalangeal joint was 85 degrees and that of the distal interphalangeal joint 48 degrees. The mean grip strength was 92% of the uninvolved hand. Twelve patients have returned to their original occupations. There has been no radiological osteolysis or clinical osteomyelitis. This modified traction system has given acceptable results with a low rate of complications. It is light, cheap, effective and easy to apply. PMID- 15125131 TI - A three-dimensional classification for fractures of the proximal humerus. AB - Existing classifications of fractures of the head of the humerus are inadequate in terms of interobserver reliability and the predictability of the clinical outcome. From a combined study of 73 fracture specimens in museums and 84 CT three-dimensional reconstructions in patients, we have devised a classification which appears to be more useful clinically. Common patterns of fracture and a plausible mechanism of injury were observed. In 3-D most proximal humeral fractures can be organised into five basic types. These correspond in some degree to the Codman/Neer classification, but differ significantly in regard to the more complex patterns of fracture. We observed a logical progression from simple to complex fractures. An interobserver reliability study was carried out which indicated the improved usefulness of this new 3-D concept in providing a common language among clinicians for classifying these injuries. When surgery is indicated, the 3-D concept is also invaluable in guiding the restitution of anatomy through either open or percutaneous means. PMID- 15125132 TI - The most severe forms of Perthes' disease associated with the homozygous Factor V Leiden mutation. AB - It has recently been postulated that thrombophilia may have a role in the aetiology of Perthes' disease. The published reports, however, remain conflicting. In this study a retrospective analysis of the coagulation parameters was made in 47 patients with Perthes' disease and the results compared with the clinical data. Five patients with Factor V Leiden mutation were found (10.6%) and surprisingly four of them had a homozygous pattern. These four patients showed the most severe form of the disease, Catterall group IV, with flattening of the entire epiphysis, involvement of the metaphysis, shortening and broadening of the femoral neck, trochanteric overgrowth and developed mushroom-shaped aspherical laterally displaced femoral heads in dysplastic acetabula. We would like to suggest that the homozygous form of Factor V Leiden mutation has some role in the clinical course of Perthes' disease and particularly its most severe form. PMID- 15125133 TI - The ferguson medial approach for open reduction of developmental dysplasia of the hip. A clinical and radiological review of 49 hips. AB - The results of the Ferguson medial approach for open reduction of developmental dysplasia of the hip (DDH) were reviewed for 49 hips with a follow-up of more than 48 months. The mean age at operation was 12.3 months (6 to 23). The mean length of clinical and radiological follow-up was 82 months (48 to 148). Three redislocations occurred. Group I avascular necrosis according to the classification of Kalamchi and MacEwen was seen in four hips, group II in two hips and group III in one hip; 92% of the hips were classified as Severin class I and II. The acetabular index and centre edge (CE) angles were within normal limits at final follow-up, but were still significantly different from the unaffected side. We conclude that the Ferguson procedure is safe and reliable for low dislocations in children aged six to 18 months. PMID- 15125134 TI - Inter- and intra-measurer error in the measurement of Reimers' hip migration percentage. AB - Reimers' hip migration percentage is commonly used to document the extent of subluxation of the hip in children with spasticity. In this study, two measurers, with six months paediatric orthopaedic experience, measured the migration percentage on 44 pelvic radiographs of children with cerebral palsy, aged between two and eight years. Unknown to the measurers, each radiograph was duplicated, giving 22 non-identical radiographs (44 hips) which were measured twice at time 0 and twice six weeks later. The intra-measurer, intra-sessional absolute differences between the first and second measurements ranged from 0% to 23%, with median values of 2.5% to 3.6%. The intra-sessional median absolute differences were not statistically different between the two measurers and measuring sessions (p = 0.42, Kruskal-Wallis test). The inter-sessional absolute differences for measurements made by the same measurers ranged from 0% to 18% with a median absolute difference of 1.7% to 3.2%. Overall, only 5% of the intra-measurer measurement differences, within and between sessions, were above 13%. Repeated measurements by one measurer over time must, therefore, vary by more than 13% in order to be 95% confident of a true change. The inter-measurer error was higher with median absolute differences between the two measurers' measurements of the same hip of 3.25% to 5% (0% to 26%) and a 95th upper confidence interval of 21% to 23%. Averaging the four separate measurements over the two sessions reduced the inter-measurer error to a median absolute difference of 2.8%, but did not improve the 95th upper confidence interval, which measured 22.4%. Such inter measurer errors may be clinically unacceptable. PMID- 15125135 TI - Long-term deterioration of joint evaluation scores. AB - We investigated the long-term changes in the Harris Hip and Knee Society scores (HSS and KSS) to determine whether they result from overall functional decline rather than actual changes in the condition of the prosthesis. The HHS for 106 total hip arthroplasties with a minimum follow-up of ten years, no medical complications after operation and no evidence of radiological loosening, and the KSS for 264 total knee arthroplasties with a minimum follow-up of 12 years and no medical complications after operation or signs of radiographical loosening were evaluated. There were statistically significant drops in the functional scoring components of the joint evaluation systems despite no loosening of the prostheses or other significant medical complications. The HHS declined at an average of 0.67 points per year from between three and ten years after operation (p < 0.0001). Contributing to this were deterioration in gait and limp (p < 0.0004), the use of support aids (p < 0.0001), the distance walked (p < 0.0001) and the ability to climb stairs (p < 0.0455). The functional component of the KSS declined significantly at an average 0.88 points per year betwen the third and 12th years (p < 0.0001). There were significant declines in every component of the functional score including the distance walked (p < 0.0001), the ability to climb stairs (p < 0.0001) and the use of support aids (p < 0.0001). The knee score component of the KSS did not decline significantly (p < 0.9750). The combination of functional and pain scores within the HHS system leads to an inaccurate decline in the entire score. The decline of HHS and Knee Society functional scores in total joint arthroplasties, in the absence of implant related problems, suggests that deterioration in the functional capacity of ageing patients is an important factor in longitudinal studies using these scoring systems. PMID- 15125136 TI - The influence on human osteoblasts in vitro of non-steroidal anti-inflammatory drugs which act on different cyclooxygenase enzymes. AB - There is increasing evidence that non-steroidal anti-inflammatory drugs (NSAIDs) can adversely affect bone repair. We have, therefore, studied the in vitro effects of NSAIDs, which differentially inhibit cyclooxygenases (COX), the prostaglandin/thromboxane synthesising enzymes, on human osteoblasts. Indomethacin and the new nitric oxide (NO)-donating NSAIDs block the activity of both COX-1 and COX-2. Indomethacin and 5,5-dimethyl-3-(3 fluorophenyl)-4-(4 methylsulphonal) phenyl-2 (5H)-furanone (DFU) reduced osteoblast numbers in a dose-dependant manner and increased collagen synthesis and alkaline phosphatase activity. The reduction in osteoblast numbers was not caused by loss of adhesion and was reversible. Neither NSAID influenced DNA synthesis. There was no difference between the effects of indomethacin and DFU. NO-NSAIDs did not affect cell numbers. These results suggest that care should be taken when administering NSAIDs to patients with existing skeletal problems and that NO-NSAIDs may be safer. PMID- 15125137 TI - The posterior cruciate ligament during flexion of the normal knee. AB - The posterior cruciate ligament (PCL) was imaged by MRI throughout flexion in neutral tibial rotation in six cadaver knees, which were also dissected, and in 20 unloaded and 13 loaded living (squatting) knees. The appearance of the ligament was the same in all three groups. In extension the ligament is curved concave-forwards. It is straight, fully out-to-length and approaching vertical from 60 degrees to 120 degrees, and curves convex-forwards over the roof of the intercondylar notch in full flexion. Throughout flexion the length of the ligament does not change, but the separations of its attachments do. We conclude that the PCL is not loaded in the unloaded cadaver knee and therefore, since its appearance in all three groups is the same, that it is also unloaded in the living knee during flexion. The posterior fibres may be an exception in hyperextension, probably being loaded either because of posterior femoral lift off or because of the forward curvature of the PCL. These conclusions relate only to everyday life: none may be drawn with regard to more strenuous activities such as sport or in trauma. PMID- 15125138 TI - The significance of radiolucent zones surrounding pedicle screws. Definition of screw loosening in spinal instrumentation. AB - We examined the radiographs from a prospective clinical study of fixation by pedicle screws and those from an experimental study in a sheep model. In the clinical study, instruments were removed from 21 patients after implantation for 11 to 16 months and the extraction torques of the screws were recorded. A structured protocol was used for the radiological examinations. In the experimental study, loaded pedicle screw instrumentations were implanted in the sheep for six or 12 weeks. After radiological examination the pull-out resistance and the histological characteristics were studied. In the clinical study, all screws with radiolucent zones had a significantly reduced mean extraction torque compared with screws without radiolucent zones (16 +/- 10 Ncm v 403 +/- 220 Ncm; p < 0.0001). In the experimental study the mean maximum pull-out resistance for the screws with radiolucent zones was significantly lower than for those with no radiolucency (243 +/- 156 N v2214 +/- 578 N; p = 0.0006) and the mean bone-to screw contact was reduced for screws with zones compared with those without zones (8 +/- 9% v 55 +/- 29%; p = 0.0002). Our findings showed that all screws with radiolucent zones had low extraction torques or low pull-out resistance. A radiolucent zone is a good indicator of loosening of a pedicle screw. PMID- 15125139 TI - Complete remodelling of displaced fractures of the neck of the phalanx. PMID- 15125140 TI - A strategy for reducing blood-transfusion requirements in elective orthopaedic surgery. PMID- 15125141 TI - Complications of arthroscopic reconstruction of the anterior cruciate ligament. PMID- 15125142 TI - Ventricular surface activation time imaging from electrocardiogram mapping data. AB - Non-invasive imaging of cardiac electrophysiology provides a non-invasive way of obtaining information about electrical excitation. An iterative algorithm based on a general regularisation scheme for non-linear, ill-posed problems in Hilbert scales was applied to the electrocardiographic inverse problem, imaging the ventricular surface activation time (AT) map. This method was applied to electrocardiographic data from a 31-year-old healthy volunteer and a 24-year-old patient suffering from a Wolff-Parkinson-White (WPW) syndrome. The objective was to evaluate non-invasive AT imaging of an autonomous sinus rhythm and to quantify the localisation error of non-invasive AT imaging by localising the accessory pathway of the WPW syndrome and a pacing site for left ventricle pacing. The distances between the invasive and non-invasive localisation of the pacing site and the accessory pathway were 8 mm and 5 mm. The clinical case presented, shows that this non-invasive AT imaging approach may enable the reconstruction of single focal events with sufficient accuracy for potential clinical application. PMID- 15125143 TI - Quantitative modelling of interaction of propafenone with sodium channels in cardiac cells. AB - A mathematical model of the interaction of propafenone with cardiac sodium channels is based on experimental data that demonstrate use-dependent effects of the drug. The Clancy-Rudy model is applied to describe Na-channels in absence of the drug. The values of rate constants of the drug-receptor reaction are fitted to experimental data by iterative computer simulations using a genetic algorithm. The model suggests the following interpretation of available experimental results: First, drug molecules have access to the binding sites predominantly in the inactivated states. Secondly, the biphasic development of the block during depolarisation is consistent with a rapid increase due to drug binding in the fast inactivated state (rate constants k(on) = 645 micromol(-1) l s(-1), k(off) = 16.21 s(-1)) and a slow increase due to binding in the intermediate inactivated state (rate constants approximately 100-fold lower), followed by transition to the drug-occupied slow inactivated state (rate constants 0.784 and 0.921 s(-1)). Thirdly, the observed biphasic time course of recovery of I(Na) from block following restoration of the resting voltage results from simultaneous relief of block from the channels residing in the intermediate and slow inactivated states. Fourthly, the accumulation of blocked channels in the slow inactivated state is responsible for the observed use-dependent effects. Fifthly, when incorporated into a quantitative description of the electrical activity of a ventricular cell, the model predicts that propafenone (0.2 micromol l(-1)) effectively suppresses premature excitations, leaving the regular action potentials nearly unaffected. PMID- 15125144 TI - Modelling study of the acute cardiovascular response to hypocapnic hypoxia in healthy and anaemic subjects. AB - The present study analyses the cardiovascular response to acute hypocapnic hypoxia (simulating the effect of respiration at high altitude) both in healthy, unacclimatised subjects and in subjects with moderate anaemia, by means of a mathematical model of short-term cardiovascular regulation. During severe hypoxia, cardiac output and heart rate (HR) exhibit a significant increase compared with the basal level (cardiac output: +90%; HR: +64%). Systemic arterial pressure remains quite constant or shows a mild increase. Coronary blood flow increases dramatically (+200%), thus maintaining a constant oxygen delivery to the heart. However, blood oxygen utilisation in the heart augments, to fulfil the increased power of the cardiac pump during hypoxia. Cerebral blood flow rises only at very severe hypoxia but, owing to the vasoconstrictory effect of hypocapnia, its increase (+80%) is insufficient to maintain oxygen delivery to the brain. The model suggests that a critical level for the aerobic metabolism in these organs (heart and brain) is reached at an oxygen partial pressure in arterial blood (PaO2) of approximately 25 mmHg. Moderate anaemia during normoxia is compensated by an increase in cardiac output (+22%), a decrease in total peripheral resistance (-30%) and an increase in O2 extraction from blood (+40%). As cardiovascular regulation mechanisms are already recruited in anaemic subjects at rest, their action soon becomes exhausted during hypocapnic hypoxia. Critical levels for vital functions are already reached at a PaO2 of approximately 45 mmHg. PMID- 15125145 TI - Anatomy- and physics-based facial animation for craniofacial surgery simulations. AB - A modelling approach for the realistic simulation of facial expressions of emotion in craniofacial surgery planning is presented. The method is different from conventional, non-physical techniques for character animation in computer graphics. A consistent physiological mechanism for facial expressions was assumed, which was the effect of contracting muscles on soft tissues. For the numerical solution of the linear elastic boundary values, the finite element method on tetrahedral grids was used. The approach was validated on a geometrical model of a human head derived from tomographic data. Using this model, individual facial expressions of emotion were estimated by the superpositioning of precomputed single muscle actions. PMID- 15125146 TI - Method for modelling cerebral blood vessels and their bifurcations using circular, homogeneous, generalised cylinders. AB - A method for automatic modelling of blood vessels and their bifurcations from 3D scans of the brain is presented. The method is a three-step procedure. First, a skeleton of the cerebral blood vessels is developed, and then the surfaces of the blood vessels are located using an active contour approach. The active contour approach uses circular homogeneous generalised cylinders (CHGCs) to model the thin, elongated blood vessels. Finally, a novel method for modelling the surfaces of the bifurcations in a vessel tree is presented. The method was tested on simulated data: a computed tomography angiography (CTA) and four magnetic resonance angiography (MRA) volumes. Furthermore, the method was tested on ten magnetic resonance images (MRIs) to demonstrate its robustness. The test on the simulated data indicated that the approach for the surface modelling of vessels had a mean radius error of less than 0.1 mm and a mean localisation error of 0.1 mm. Surface models evaluated by an expert in vascular neurosurgery were found to have a smooth appearance and generally agreed with the image data. The test on the MRI scans indicated that the method performed well in noisy environments. PMID- 15125147 TI - Pilot point temperature regulation for thermal lesion control during ultrasound thermal therapy. AB - The fundamental goal of ultrasound thermal therapy is to provide proper thermal lesion formations for effective tumour treatment. The quality of the therapy depends mostly on its positional precision. To date, most ultrasound thermal therapy treatments have focused on the formation of power or temperature patterns. The non-linear and time-delay effects of thermal dose formation prohibit direct control of the thermal dose distribution. In the paper, the control of thermal lesions by regulation of the temperature of a pilot point is proposed. This scheme utilises the high correlation between temperature elevation and thermal dose at the forward boundary of thermal lesions. To verify the feasibility, a 2D ultrasound phased array system was used to generate thermal lesions of various sizes, and the temperature elevation required to generate a thermal dose threshold was investigated. Results showed that the required temperature elevation was found to be a reasonably constant value of 52.5 degrees C under differing conditions when the focal area was small. When the focal area under consideration was large, the required temperature elevation became a monotonic function of blood perfusion rate, ranging from 49.2 to 52.5 degrees C. When the reference temperature of the pilot point was set at a conservative value (52.5 degrees C), the thermal lesions were controlled precisely under a wide range of blood perfusion and power pattern changes, tested by using a more realistic model that takes into account thermal-induced attenuation and blood perfusion changes. This changed the complex thermal dose control problem into a simple temperature regulation problem, which makes implementation of thermal lesion control easier, giving the scheme a high potential for application to current ultrasound thermal therapy systems. PMID- 15125148 TI - Wavelet-based statistical approach for speckle reduction in medical ultrasound images. AB - A novel speckle-reduction method is introduced, based on soft thresholding of the wavelet coefficients of a logarithmically transformed medical ultrasound image. The method is based on the generalised Gaussian distributed (GGD) modelling of sub-band coefficients. The method used was a variant of the recently published BayesShrink method by Chang and Vetterli, derived in the Bayesian framework for denoising natural images. It was scale adaptive, because the parameters required for estimating the threshold depend on scale and sub-band data. The threshold was computed by Ksigma2/sigma(x), where sigma and sigma(x) were the standard deviation of the noise and the sub-band data of the noise-free image, respectively, and K was a scale parameter. Experimental results showed that the proposed method outperformed the median filter and the homomorphic Wiener filter by 29% in terms of the coefficient of correlation and 4% in terms of the edge preservation parameter. The numerical values of these quantitative parameters indicated the good feature preservation performance of the algorithm, as desired for better diagnosis in medical image processing. PMID- 15125149 TI - Real-time identification and archiving of micro-embolic Doppler signals using a knowledge-based DSP system. AB - Identification of micro-emboli in the cerebral circulation using transcranial Doppler ultrasound provides valuable clinical information, but, currently, embolic signal detection and analysis are significantly limited because they mainly rely on costly off-line analysis by human experts. In this study, a reliable, high-resolution, real-time automated system for the detection and archiving of embolic signals was designed and implemented using expert system theory and modern DSP technology. Preliminary tests were conducted to evaluate the functions and the performance of the system using data from ten carotid endarterectomy patients and two normal volunteers. Using the widely accepted 7 dB threshold for human reliability and a human expert, majority-decision gold standard, the real-time system reached sensitivity and specificity of 93.6% and 99.3%, respectively, which were close to the results obtained by three human experts under ideal laboratory conditions (90.1% and 99.8%, 98.4% and 99.9%, 98.9 and 99.9%). The new system has the potential to be used either as a bedside monitoring and signal acquisition device, or as a laboratory investigation tool. PMID- 15125150 TI - Identification of the breast boundary in mammograms using active contour models. AB - A method for the identification of the breast boundary in mammograms is presented. The method can be used in the preprocessing stage of a system for computer-aided diagnosis (CAD) of breast cancer and also in the reduction of image file size in picture archiving and communication system applications. The method started with modification of the contrast of the original image. A binarisation procedure was then applied to the image, and the chain-code algorithm was used to find an approximate breast contour. Finally, the identification of the true breast boundary was performed by using the approximate contour as the input to an active contour model algorithm specially tailored for this purpose. After demarcation of the breast boundary, all artifacts outside the breast region were eliminated. The method was applied to 84 medio-lateral oblique mammograms from the Mini-MIAS database. Evaluation of the detected breast boundary was performed based upon the percentage of false-positive and false negative pixels determined by a quantitative comparison between the contours identified by a radiologist and those identified by the proposed method. The average false positive and false negative rates were 0.41% and 0.58%, respectively. The two radiologists who evaluated the results considered the segmentation results to be acceptable for CAD purposes. PMID- 15125151 TI - Mathematical model for pre-operative planning of linear and closing-wedge metatarsal osteotomies for the correction of hallux valgus. AB - First-metatarsal osteotomy is performed for an abnormal intermetatarsal angle and severe hallux valgus deformity. The metatarsal head is slid along the osteotomy and fixed with a Kirschner wire to achieve angular and linear realignment with respect to the other metatarsals. When reduction of joint subluxation is also sought, a bone wedge is removed. A mathematical model and a corresponding computer-based tool were developed for pre-operative planning of the optimum surgical solution. Standard intermetatarsal angle correction, head-to-sesamoids re-alignment, joint subluxation reduction and metatarsal length adjustment were all pursued. A standard pre-surgical dorso-plantar radiograph of the patient, with an additional metal ball for calculation of X-ray magnification, was necessary to analyse the original and final geometrical configurations. For the first time, expected bone loss and wire diameter were also considered. The angles of bone cuts and the amount of sliding along these cuts were calculated by the model according to the surgical goal. All the resulting geometrical parameters, both in the pre- and planned post- operative configuration, were provided in graphical and tabular form. In a change from former qualitative analysis of radiograms, the surgeon can now enter the operating theatre with a detailed plan of the goal to be achieved. PMID- 15125152 TI - Biomechanical modelling of segmental instrumentation for surgical correction of 3D spinal deformities using Euler-Bernoulli thin-beam elastic deformation equations. AB - A simplified computer-modelling technique intended to analyse 3D spinal deformity correction with segmental instrumentation is presented. The spine was modelled as a thin beam-composed structure linked by implants to two deformable rods. The Landau vector representation of Euler-Bernoulli beam elastic deformation equations was used to formulate the simulation approach. All types of essential deformation (bending, torsion, tension, compression) were considered. An iterative numerical method was proposed to obtain an appropriate load, able to deform the spine axial curve to the desired post-operative shape. A simulation based on the spine of a real scoliotic patient (thoracic and lumbar Cobb angles: 39 degrees and 8 degrees), corrected using surgical instrumentation intervention, is presented. Force loads within the range of 20-350 N were able to deform the pre-operational spine axial curve to the post-operational one with a root mean square approximation error of 3.7 mm. Similarly good corrections were obtained using different force patterns. This highlights the uncertainty of which corresponding surgical instrumentation to use. Such uncertainty is related to the 'ill-posed problems' property of mechanical systems. PMID- 15125154 TI - Three-dimensional stress analysis for the mechanics of plantar ulcers in diabetic neuropathy. AB - In diabetic neuropathic subjects, the hardness of foot sole soft tissue increases, and its thickness reduces, in different foot sole areas. Finite element analysis (FEA) of a three-dimensional two-arch model of the foot was performed to evaluate the effect of foot sole stresses on plantar ulcer development. Three sets of foot sole soft-tissue properties, i.e. isotropic (with control hardness value), diabetic isotropic (with higher hardness value) and anisotropic diabetic conditions, were simulated in the push-off phase, with decreasing foot sole soft-tissue thicknesses in the forefoot region, and the corresponding stresses were calculated. The results of the stress analyses for diabetic subject (anisotropic) foot models showed that, with non-uniformly increased hardness and decreased foot sole soft-tissue thickness, the normal and shear stresses at the foot sole increased (compared with control values) by 52.6% and 53.4%, respectively. Stress analyses also showed high ratios of gradients of normal and shear stresses of the order of 6.6 and 3.3 times the control values on the surface of the foot sole, and high relative values of stress gradients for normal and shear stresses of 6.25 and 4.35 times control values, respectively, between the foot sole surface and the adjacent inner layer of the foot sole, around a particular region of the foot sole with anisotropic properties. These ratios of high gradients and relative gradients of stresses due to changes in soft-tissue properties may be responsible for the development of plantar ulcers in diabetic neuropathic feet. PMID- 15125153 TI - Effect of the initial implant fitting on the predicted secondary stability of a cementless stem. AB - A numerical model able to investigate the influence of biomechanical factors on the long-term secondary stability of implants would be extremely useful for the design of new cementless prosthetic devices. A purely biomechanical model of osseo-integration has been developed, formulated as a rule-based adaptation scheme. Due to its complexity, the problem was divided into three steps: preliminary implementation of the model (proof of concept); implementation of the complete model and investigation of the model solution; and model validation. The paper describes the first of these three steps. The model was implemented as a discrete-states machine, and the few parameters required were derived from the literature. It was then applied to a real clinical case. The study was conducted using the frictional contact finite element model of a human femur implanted with a cementless anatomical stem. A stable solution was achieved after between three and 15 iterations for all initial positions considered. Similar initial conditions yielded similar final configurations. The model predicted all initial configurations, with the exception of a partial osseo-integration, ranging between 62% (distal fit) and 78% (proximal fit) of the viable interface. This is in good agreement with the values reported in the literature that never exceed 75%, even in the best conditions, and report better clinical results for proximal fit. For the varus configuration, which lacks cortical support, the algorithm predicted a completed loosening. PMID- 15125155 TI - Identifying patterns of spatial current dispersion that characterise and separate the Brugada syndrome and complete right-bundle branch block. AB - The aim of the study was to detect patterns of spatial-current distribution in the late QRS and early ST-segments that distinguish Brugada-syndrome cases from complete right-bundle branch block (CRBBB). Magnetocardiograms (MCGs) were recorded from Brugada-syndrome patients (n = 6), CRBBB patients (n = 4) and the members of a control group (n = 33). The current distributions at six time points from Q-onset were estimated by producing current-arrow maps (CAMs). The angle of the current arrow of maximum amplitude at each time point was calculated. In the Brugada cases, the characteristic ST elevation was seen above the upper right chest, and abnormal currents appeared to be present in the right-ventricular outflow tract (RVOT). The angles of the abnormal arrows were -78 degrees +/- 51 degrees at 100 ms and -50 degrees +/- 61 degrees at 110 ms. In the cases of CRBBB, wide S- and R-waves were recorded above the upper right and lower right chest, respectively. The angles of the abnormal arrows for CRBBB were 152 degrees +/- 19 degrees at 100 ms, 159 degrees +/- 20 degrees at 110 ms, and 157 degrees +/- 19 degrees at 120 ms. The findings suggest that an abnormal current from the RVOT to the upper left chest may be a feature of the Brugada syndrome, and that the direction of this current is completely different from that seen in CRBBB. PMID- 15125156 TI - Fibre-optic array for curvature assessment: application in otitis diagnosis. AB - A contact-free sensor consisting of two parallel optical-fibre arrays was designed to assess surface shapes of diffusely scattering media. By sequentially illuminating objects using one fibre array and detecting the diffusely back scattered photons by the other, a source-detector intensity matrix was formed, where the matrix element (i, j) was the intensity at detector j when light source i was excited. Experimental data from convex and concave polyacetal plastic surfaces were recorded. A mathematical model was used for simulating source detector intensity matrices for the surfaces analysed in the experiments. Experimental results from the system were compared with the theoretically expected results provided by the mathematical model. The shape and relative amplitude showed similar behaviour in the experiments and simulations. A convex/concave discriminator index D, representing the detected intensity difference between two source-detector separations, was defined. The relative dynamic range of D, defined as the difference between the maximum and the minimum divided by the mean of the index, was 1.37 for convex surfaces and 0.68 for concave surfaces, at a measuring distance of 4.5 mm. The index D was positive for convex surfaces and negative for concave surfaces, which showed that the system could distinguish between convex and concave surfaces, an important result for the diagnosis of otitis media. PMID- 15125157 TI - Detection of the third heart sound using a tailored wavelet approach. AB - The third heart sound is normally heard during auscultation of younger individuals but disappears with increasing age. However, this sound can appear in patients with heart failure and is thus of potential diagnostic use in these patients. Auscultation of the heart involves a high degree of subjectivity. Furthermore, the third heart sound has low amplitude and a low-frequency content compared with the first and second heart sounds, which makes it difficult for the human ear to detect this sound. It is our belief that it would be of great help to the physician to receive computer-based support through an intelligent stethoscope, to determine whether a third heart sound is present or not. A precise, accurate and low-cost instrument of this kind would potentially provide objective means for the detection of early heart failure, and could even be used in primary health care. In the first step, phonocardiograms from ten children, all known to have a third heart sound, were analysed, to provide knowledge about the sound features without interference from pathological sounds. Using this knowledge, a tailored wavelet analysis procedure was developed to identify the third heart sound automatically, a technique that was shown to be superior to Fourier transform techniques. In the second step, the method was applied to phonocardiograms from heart patients known to have heart failure. The features of the third heart sound in children and of that in patients were shown to be similar. This resulted in a method for the automatic detection of third heart sounds. The method was able to detect third heart sounds effectively (90%), with a low false detection rate (3.7%), which supports its clinical use. The detection rate was almost equal in both the children and patient groups. The method is therefore capable of detecting, not only distinct and clearly visible/audible third heart sounds found in children, but also third heart sounds in phonocardiograms from patients suffering from heart failure. PMID- 15125158 TI - Measurement of the skin microcirculation through intact bandages using laser Doppler flowmetry. AB - The microcirculation under compression bandages has been assessed by numerous methods; however, the measurement techniques can disrupt the bandage-skin interface, affecting the measurement. In this study, a non-invasive method for measuring cutaneous blood flow using laser Doppler flowmetry (LDF) is presented. Ten volunteers had their microcirculation assessed by a laser Doppler probe being placed on their upper forearm with and without a light-transmissive gel and with a compression bandage plus light-transmissive gel. A circulatory challenge to the bandaged forearm in two of the volunteers was also undertaken. The median (95% confidence interval) perfusion (p.u.) for the skin surface was 24 (15-33) perfusion units (p.u.), and the skin plus light-transmissive gel demonstrated a higher perfusion: 66 (50-82) p.u., (p < 0.012). The addition of the compression bandage, with additional gel allowed to permeate through to the underlying skin, decreased the perfusion to 27 (20-34) p.u. (p < 0.007). In both volunteers, the microcirculatory flow responded to the vascular challenge, resulting in flow changes related to the cuff pressure (45-27 and 14-8 p.u.). This method demonstrated that it may be possible to assess the microcirculation through intact bandages, without the need to place any sensors at the skin-bandage interface. PMID- 15125159 TI - Integration-plot test for peri-stimulus time histograms in human motor units. AB - A statistical test is proposed for peri-stimulus time histograms in human motor units for the case where a test stimulus is delivered at a constant interval after a previous discharge. This mathematically described test included the notion of the multiple comparison and thus achieved higher sensitivity than the previously proposed method. With regard to data acquisition, the interval from a sham stimulus to the next discharge was acquired as a control, and the total number of samples was set to be four times as large as that in the test situation to reduce the statistical scattering noise. A newly defined statistical object, the integration plot (timewise accumulation of the test histogram without control subtraction) was used for this statistical test. The integration plot had less noise than the cumulative sum (CUSUM) plot (1/square root(2) in theory) and thus represented the neural effect. To compare this integration-plot test with that of the CUSUM, a simulation experiment that compared two sample histograms (one of which had a faint structural change from 20 ms) was conducted. As a result, the present test succeeded in detecting the onset of the change point earlier (23 ms on average) than the CUSUM test (27 ms on average), and the detection probability was also higher (9 out of 10) than the CUSUM (6 out of 10). It was therefore confirmed experimentally that the present statistical test had higher sensitivity than that of the CUSUM proposed previously. PMID- 15125160 TI - Two-electrode low supply voltage electrocardiogram signal amplifier. AB - Portable biomedical instrumentation has become an important part of diagnostic and treatment instrumentation, including telemedicine applications. Low-voltage and low-power design tendencies prevail. Modern battery cell voltages in the range of 3-3.6 V require appropriate circuit solutions. A two-electrode biopotential amplifier design is presented, with a high common-mode rejection ratio (CMRR), high input voltage tolerance and standard first-order high-pass characteristic. Most of these features are due to a high-gain first stage design. The circuit makes use of passive components of popular values and tolerances. Powered by a single 3 V source, the amplifier tolerates +/- 1 V common mode voltage, +/- 50 microA common mode current and 2 V input DC voltage, and its worst-case CMRR is 60 dB. The amplifier is intended for use in various applications, such as Holter-type monitors, defibrillators, ECG monitors, biotelemetry devices etc. PMID- 15125161 TI - Staff ratios: California law may spread to other states. PMID- 15125162 TI - Pica. An underappreciated cause of electrolyte abnormalities. PMID- 15125163 TI - The role of the nephrology social worker in team training for patient nonadherence. PMID- 15125164 TI - Can we revitalize the ESRD program? PMID- 15125165 TI - Resolving the nursing crisis. Ratios, faculty shortage, and a demand that keeps growing. PMID- 15125166 TI - The role of physician assistants in improving renal care. AB - Currently, nephrology PAs remain a small group. According to 2003 census data from The American Academy of Nephrology Physician Assistants, only 98 of 20,646 survey respondents identified themselves as practicing in nephrology. The future of PAs or nurse practitioners in nephrology is not only very bright, but is also an absolute necessity. We have known for many years that the number of individuals with kidney disease in the United States is increasing at a rate that outpaces our ability to develop and train nephrologists. This has resulted in an ever-increasing ratio of patients to clinical nephrologists. The workload for management of dialysis patients on a daily basis is becoming exhaustive and will not improve. The fastest growing segment of dialysis patients is now people in their 70s and 80s, and they bring with them multiple chronic health problems that are affected by dialysis and the treatment of their renal disease. The result is the need for closer monitoring, not less. The role of physician extenders can have a very positive impact for this patient population. Being the eyes, ears, nose, and fingers of our nephrologists can help in avoiding potential major problems in the outpatient arena. There is not a magic formula in caring for this patient population; it is a matter of spending time and becoming familiar with our patients, a premium most nephrologists do not have at present. It is not a matter of willingness; it is a matter of capability, of being in more than one place, and of having time to make the patient assessments. I think there is a great opportunity for nephrologists to create a new segment of providers to assist them in these endeavors. They can sponsor PAs as preceptors before graduation so that the students can have the opportunity to see what it takes to care for this population, the level of medicine they need to learn, and the responsibility they will need to accept. The nephrologist will benefit from working with a PA that has a good foundation of medical education that the nephrologist can tailor to his or her own method of practice in order to become comfortable in the relationship long-term. The future of care for our end-stage renal disease population needs all our efforts to succeed. PMID- 15125167 TI - Workplace resources, patient caseloads, and job satisfaction of renal social workers in the United States. A Survey/Part 1. PMID- 15125168 TI - Leg training. Accessing the body's energy reserves. PMID- 15125169 TI - 10 ways to improve patient relations. PMID- 15125170 TI - Scope of medical colleges in private sector. AB - Medical profession is still the most sought after profession, there are thousands of bright students who pass their F.Sc examination in First Division and desire to join medical colleges but are unable to get admission in government medical colleges. There is a great shortage of doctors in the country. Hundreds of young students go abroad for medical education, costing huge sum of foreign exchange to the families and to the government spending 5 to 7 years and obtaining degrees which are not recognized in Pakistan. Therefore to fulfill the demand of the public, and to overcome the shortage of doctors in the country and to save the valuable foreign exchange, private sector medical colleges are much needed in Pakistan. However, establishment of medical colleges is very expensive and difficult. PMDC is very firm in implementing its rules and regulations. Those colleges which fulfill the laid down requirement of PMDC, help in overcoming the shortage of doctors in the country and provide much needed health care to the population. Such medical colleges are much appreciated by the public and by the govt. Such private sector medical colleges have great future and some of them may become famous and outstanding like the private sector medical colleges in the USA. PMID- 15125171 TI - Accuracy of urine dipstick to predict urinary tract infections in an emergency department. AB - BACKGROUND: Urine dipstick is a useful and commonly used test in the Emergency Department because of its rapidity and low cost; however its diagnostic accuracy is debatable. Our objective was to compare the urine dipstick and urinalysis for Urinary Tract Infection in a developing country, where there are significant cost considerations. METHODS: This was an observational study of adults' patients presenting to Section of Emergency section (SEM) of the Aga Khan University Hospital, from March to May 1998. The patient's urine sample was tested immediately, using the Multistix 10SG. The sample was sent within one hour to the hospital laboratory for analysis, while the urinary specimen was sent for culture, where appropriate. The dipstick results were compared with the results of automated urinalysis in the laboratory, leukocyte counts on microscopy and urine culture. Sensitivity, Specificity, and predictive values were also calculated. RESULTS: We evaluated 984 samples of urine during the study period. The sensitivity of nitrite test was 81% and that of leukocyte esterase 77% for positive cultures. However, the sensitivity for combined nitrite and leukocyte esterase test was 94%. Nitrite test was more specific (87%) than leukocyte esterase test (54%) or both tests taken together (50%). The predictive value of nitrite and leukocyte esterase together for a negative urine culture was 95%. Leukocyte esterase test sensitivity increased as the number of white blood cells on microscopy increased. Similarly the predictive value of leukocytes on microscopy for a positive culture increased as the number of leukocytes increased. CONCLUSION: Dipstick alone cannot accurately predict urinary tract infection in emergency department. PMID- 15125172 TI - Patterns of pancytopenia patients in a general medical ward and a proposed diagnostic approach. AB - BACKGROUND: There has been little systematic study on the clinical spectrum of pancytopenia. This study was done to describe the etiology, presentation and outcome of patients with pancytopenia presenting in a general medical ward. METHODS: Hundred patients with pancytopenia were included in the study from October 2001 to October 2002. Patients on cancer chemotherapy were excluded. Blood counts, bone marrow examinations and trephine biopsies were performed according to standard methods. RESULTS: In all cases, megaloblastic anemia constituted the largest group (n = 39), and also seen in conjunction with hemolytic anemia and septicemia. Hypersplenism secondary to portal hypertension (cirrhosis) was the second most common diagnosis (n = 19). Aplastic anemia, septicemia and myelodysplasia were other common causes. Two patients were the suspected cases of viral hemorrhagic fever. Thirteen (13%) patients expired. Absolute neutrophil count (ANC) less than 500/microliter was seen in 14 (14%) patients, among which 6 (15.3%) had megaloblastic anemia, 3 (37.5%) had aplastic anemia, and 2 (40%) had myelodysplasia. Eleven patients with platelet counts < or = 10 x 10(9)/L, 6 (54.5%) presented with bleeding; and 2 of these 8 had aplastic anemia and 1 patient with megaloblastic anemia. MCV values > 100 fL and > 110 fL were more frequent in patients with megaloblastic anemia with most prominent anisopoikilocytosis, microcytosis and fragmented RBCs. Macrocytosis was noted in 35 (89.7%) patients with megaloblastic anemia and 12 (63.1%) with hypersplenism, 4 (50%) with aplastic anemia. Hypersegmented neutrophils were noted in the blood films of 36 (92.3%) patients with megaloblastic anemia. CONCLUSION: Megaloblastic anemia, hypersplenism and aplastic anemia are the common causes of pancytopenia in our study. PMID- 15125173 TI - Presentation and management of empyema thoracis at Lady Reading Hospital Peshawar. AB - BACKGROUND: Empyema thoracis remains a common thoracic problem with challenging management strategies. We undertook the present study to outline key aspects of the presentation and management of this condition at our tertiary care hospital. METHODS: We analyzed 105 consecutive patients treated for empyema thoracis over a one-year period at Thoracic Surgical unit, Lady Reading Hospital Peshawar Pakistan. The study included patients aged 10-60 years of either sex. Patients were subjected to detailed diagnostic and management protocols with a view to define successful diagnostic and management strategies. RESULTS: The majority of patients (68%) were male, with a mean age of 28.5 +/- 14.2 years, a majority (42%) being in the 10-20 years age group. Common presentation was with fever (73%), cough (65%) and chest pain (60%). The mean duration of symptoms was 6.1 weeks. Common aetiologies of empyema were pneumonia (46.7%), iatrogenic (21.9%), traumatic (16.2%) and malignancies (11.4%). Forty patients (38%) underwent an unsuccessful therapeutic procedure prior to admission to the Thoracic unit. In the unit, 58 patients received closed intercostal drainage (31 of which required further intervention), five patients were treated with repeated thoracentesis and 8 patients were subjected to fibrinolytic therapy. The majority of patients underwent a surgical procedure like rib resection (7), decortication (23), thoracoplasty (3) and other procedures in the first instance with only 4 patients requiring further surgery. Majority of patients (97/105, 92.4%) were cured of their disease. The hospital mortality was 7.6%. CONCLUSION: Multiple therapeutic options exist for the treatment of thoracic empyema. Optimal therapy requires selection of the most appropriate first procedure for each patient with post procedure imaging to avoid inordinate delays between interventions. Early referral of all empyema patients to thoracic units for definitive therapy is recommended. PMID- 15125174 TI - Bloodstream infections in febrile neutropenic patients: bacterial spectrum and antimicrobial susceptibility pattern. AB - BACKGROUND: Bacterial infections are the major cause of morbidity and mortality among neutropenic patients. Prompt administration of empiric antimicrobial therapy for febrile neutropenic patients is considered vital. Before putting neutropenic patients on empiric antimicrobial regimens, it is essential to be aware of the spectrum of locally prevalent pathogens and their susceptibility pattern. METHODS: We studied the bacterial spectrum and antimicrobial susceptibility pattern of organisms causing bloodstream infections in febrile neutropenic patients in Armed Forces Bone Marrow Transplant Centre, Rawalpindi and the Department of Oncology, Combined Military Hospital, Rawalpindi over a period of nine months from January to September 2002. RESULTS: Blood specimens for culture and susceptibility testing were collected from 158 febrile patients with neutropenia. Eighty-three organisms were isolated from 60 patients. Thirty six (43%) isolates were Gram-positive cocci and forty-seven (57%) were Gram negative rods. Among the Gram-positive cocci, coagulase negative staphylococci (CoNS) were the predominant pathogens (26%), followed by Staphylococcus aureus (8%). Among Gram-negative rods, Escherichia coli was the predominant isolate (13%) followed by Klebsiella pneumoniae (10%). Acinetobacter johnsonii (10%) and Pseudomonas aeruginosa (7%). Nine specimens yielded polymicrobial growth. Forty percent of Staphylococcus aureus and 55% of CoNS were resistant to methicillin. All the Gram-positive isolates were susceptible to vancomycin and teicoplanin. Among the Gram-negative rods, there was 100% resistance to ampicillin, 65% to gentamicin, 47% to amikacin and 66% to third generation cephalosporins. All the gram-negative isolates were susceptible to imipenem. CONCLUSION: The spectrum of isolates among febrile neutropenic patients in our population appears to be shifting towards Gram-positive microorganisms. Due to increasing levels of drug resistance among the isolates, a glycopeptide in combination with a carbapenem would be a prudent choice as empiric therapy in high-risk cases. PMID- 15125175 TI - Comparison of job satisfaction and stress among male and female doctors in teaching hospitals of Karachi. AB - BACKGROUND: Job dissatisfaction and stress among doctors affect the quality of health care. We assessed the levels of satisfaction for workplace characteristics and job stress among doctors of three teaching hospitals in Karachi. METHODS: A postal-survey was conducted between May to June 2002. Satisfaction for workplace characteristics and stress were inquired and graded by Likert scale (1 = very low to 5 = very high). Questions were also asked about its affect on their lives. In order to calculate the differences in means of job satisfaction and stress score by sex of doctors, chi-square and t-test with 95% confidence interval (CI) were used. Out of 270 doctors approached conveniently, 189 (70%) responded and 182 questionnaires were found complete for analysis. RESULTS: Majority (68%) of the doctors was not satisfied with their jobs. Overall, the mean scores for satisfaction were low for workplace characteristics. Lowest scores were found for pay and benefits 2.12 (SE 0.8), safety and security 2.15 (SE 0.8) and workload 2.69 (SE 0.9). Female doctors had significantly lower satisfaction about workload (mean job satisfaction score difference = 0.60; 95%CI, 0.24-0.97), relation with colleagues (mean job satisfaction score difference = 0.49: 95%CI, 0.11-0.87) and autonomy (mean job satisfaction score difference = 0.45; 95%CI, 0.07-0.82) as compared to their male counterparts. Overall, 48% of doctors graded job stress from high to very high levels. CONCLUSION: Majority of doctors working at these teaching hospitals of Karachi had poor satisfaction level for workplace characteristics and higher levels of job stress. This suggests that immediate steps should be taken for their control and management. This study invites further research to explore, implement and evaluate intervention strategies for prevention of stress and improvement in job satisfaction. PMID- 15125176 TI - High frequency of false positive results in HIV screening in blood banks. AB - BACKGROUND: This study was carried out to determine the frequency of false- positive results during serological screening for the presence of antibodies against HIV-1/2 in blood banks. METHODS: A cross-sectional study was conducted from January--December 1999 as screening of voluntary non-remunerated blood donor pool for HIV in the public sector blood banks, in all the six divisions of Balochistan. 5000 subjects were screened for the presence of antibodies against HIV-1/2. The subjects were all males between the age group 18-50 years, attending the public sector blood banks as non-remunerated blood donors. Strategy I was adopted for initial screening, Strategy II and III were observed in retesting on ELISA, as recommended by UNAIDS/WHO for blood banks. RESULTS: Out of 5000 subjects, 48 (0.96%) were positive for HIV-1/2 on Strategy I, 37 (77% of 48) met the criteria of false positive, while only 11 (0.22% of 5000) were found to be true positive. CONCLUSION: In blood banks, screening for HIV antibodies is performed for intervention of the positive donations. UNAIDS/WHO Strategy-I is observed on a smaller workload blood banks where donations are less than 20 per day. A high rate of false positive results in serological HIV screening on Strategy-I depicts that the test is highly sensitive but not highly specific. Labeling someone with HIV positive, when actually he is not, forces the health authorities to find other ways of HIV screening in blood banks, which should be much more specific and therefore reliable. PMID- 15125177 TI - Relative frequency of hepatitis B and C viruses in patients with hepatic cirrhosis at DHQ Teaching Hospital D. I. Khan. AB - BACKGROUND: Hepatic cirrhosis is an important cause of morbidity and mortality worldwide. It is generally considered irreversible and premalignant condition. However reversibility of the disease has been demonstrated in some cases after removal of the cause. There are various causes leading to cirrhosis but viral cause is the most common cause worldwide. In developing countries its prevalence is more where poor knowledge about viral infections is the main cause of spread. Mode of spread and sequel of Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) are almost similar. The present study was designed to know the relative frequency of both viruses in patients with cirrhosis. METHOD: This study was conducted in the Medical unit of District Teaching Hospital of Dera Ismail Khan, from the first of May to 30th November 2002. Cirrhotic patients were admitted to Medical ward and necessary investigations were done on blood specimen for HBV and HCV. Immunoassay chromatographic technique one-step device (ACON Laboratories, USA) was applied for its viral screening. At the end 60 cirrhotic patients were analyzed for findings and comparison with other studies. RESULTS: Out of 60 patients, 28 (46.67%) were found infected with HBV, 8(13.33%) with HCV, 24 (40%) with no markers of either two viruses. One case (0.16%) was found positive for both viruses. CONCLUSION: Cirrhosis liver due to HBV is more common than HCV in this region. Males are affected more than females. PMID- 15125178 TI - Frequency of extended spectrum beta lactamase producing gram negative bacilli among clinical isolates at clinical laboratories of Army Medical College, Rawalpindi. AB - BACKGROUND: This study was carried out in Microbiology department of Army Medical College, Rawalpindi to find out the frequency of extended spectrum beta lactamase producing gram negative bacilli among clinical isolates recovered from clinical specimens received from Military Hospital, Rawalpindi. METHODS: This study was carried out from 1st Jan 2002 to 30th Dec 2002. A total of 812 consecutive Gram negative bacilli were recovered during the study period from various samples including urine, blood, pus, sputum, high vaginal swab (HVS), aspirates, i/v canula/Central venous lines (CVP), chest tubes and catheter tips. Extended spectrum beta lactamase detection in these isolates was carried out by Kirby Bauer disc diffusion method on Mueller Hinton agar. A susceptibility disk containing amoxicillin-clavulanate was placed as the inhibitor of beta lactamase in the center of the plate, and cefotaxime, ceftazidime, ceftriaxone and aztreonam disks were placed 30 mm (center to center) from the amoxicillin clavulanate disk. Enhancement of the zone of inhibition of the oxyimino-beta lactam caused by the synergy of the clavulanate in the amoxicillin-clavulanate disk was considered as evidence of ESBL production. Escherichia coli ATCC 25922 and K. pneumoniae ATCC 700603 were used as control strains. RESULTS: The frequency of ESBL producing gram negative bacilli among the clinical isolates was 45%. PMID- 15125179 TI - Waist hip ratio as an index for identifying women with raised TC/HDL ratios. AB - BACKGROUND: Obesity cannot be described solely as fat mass but the location of fat deposition is very important to determine the relation between obesity and disease. Abdominal type of obesity is linked to risk factors of atherosclerosis and to metabolic disease. Waist Hip Ratio (WHR) is a practical, simple and non invasive index of adipose tissue distribution. METHODS: We looked for a relation between WHR and TC/HDL-C ratios of a group of postmenopausal women. All the subjects in each major group were of comparable age and BMI, but the WHR varied from subject to subject. Each group was sub-divided into three tertiles based upon the WHR. Serum total cholesterol (TC) and HDL cholesterol (HDL-C) values were determined for all the subjects and TC/HDL-C ratio, which is a recognized CHD risk screening index was calculated. RESULTS: The WHR distribution was significantly different in postmenopausal women from the pre-menopausal controls, and most of the postmenopausal women showed android type of obesity with no subject in the tertile with least WHR. It was noted that the TC/HDL-C ratio increased in direct proportion with the increase in WHR and its mean value was above the desirable value in both the tertiles of postmenopausal women. The test performance characteristics of WHR showed that WHR is a sensitive and specific index for screening of high TC/HDL-c ratio. CONCLUSION: We conclude that WHR (at a cut-off point of 0.84) can serve as a sensitive and specific outpatient screening index to detect postmenopausal women with an elevated TC/HDL-C ratio. PMID- 15125180 TI - Frequency and aetiology of pruritus in admitted patients in a medical ward of Mayo Hospital, Lahore. AB - BACKGROUND: Prutitis or itching is a common complaint. It can be due to many dermatological and medical illnesses. This study was conducted to find out presence of pruritus in different diseases in patients admitted in a medical ward and to determine whether pruritus is more common due to medical causes or dermatological causes. METHODS: All patients admitted in North Medical Ward, Mayo Hospital, Lahore during March 2002 to November 2002 were included in the study. Complete medical history especially history of pruritus was taken. Physical examination was performed to diagnose the primary illness and to find cause of pruritus when present. RESULTS: 740 patients were included in the study. Sixty (8.1%) patients complained of pruritus. 5.4% had pruritus due to skin conditions. 2.7% had pruritus due to medical illnesses. CONCLUSION: The patients admitted in medical ward are having pruritus mainly due to medical diseases but skin conditions should not be ignored. PMID- 15125181 TI - Types of brain tumors in Hazara division and northern areas of Pakistan. AB - BACKGROUND: Early diagnosis and management of brain tumors is important to reduce morbidity and mortality. The objective of this study was to calculate frequency of different types of brain tumors in patients operated and managed at Ayub Teaching Hospital, Abbottabad. METHODS: This study was carried out at Neurosurgical Department of Ayub Teaching hospital, Abbottabad from September 1999 to October 2003. Records of patients who consented for surgery after CT scan evidence of brain tumors, which were proved on biopsy later on, were included in the study. The tissue biopsy was taken by open craniotomy or by computerized stereotactic method. Pattern of different brain tumors was studied during this period. RESULTS: Our study revealed predominantly tumors arising from neuroepithelium (49.2%) while tumors arising from meninges were relatively low (23.3%). Nerve sheath tumors (4.2%), germ cell tumors, mal-developmental origin (2.5%), blood vessel tumors (5.8%) and miscellaneous tumors (6.6%) were found relatively in a very low frequency. CONCLUSION: Evaluation of brain tumor frequency will help in future management plan that may be in the form of open craniotomy, stereo tactic radiosurgery, whole brain radiotherapy or gamma knife surgery. PMID- 15125182 TI - A survey of urinary bladder injuries in Abbottabad. AB - BACKGROUND: Injury of urinary bladder is not very uncommon. It has iatrogenic and non iatrogenic causes. This study was designed to determine the pattern, mode of diagnosis and management of urinary bladder injuries presenting at Ayub Teaching Hospital, Abbottabad. METHODS: Ten years hospital record of urinary bladder injuries reporting at Urology, Surgical and Gynaecology wards of Ayub Teaching Hospital, Abbottabad was analysed. The cause of injury, associated injuries, diagnostic method and management protocols were analyzed. RESULTS: During 10 years period 260 patients were treated for bladder injuries. The age range was from 5-75 years while there were more males than females. In 35% the injuries were due to RTA, in 20% by a fall from a height, in 10% by a gun shot wound and in the remaining 35% the injury was iatrogenic. Gynaecological procedures were the major cause for iatrogenic bladder injuries. CONCLUSIONS: Most of the bladder injuries seen at our hospital are due to road traffic accidents or iatrogenic causes. Both of these factors can be minimized by taking appropriate steps to improve road safety and operative procedure safety respectively. PMID- 15125183 TI - Comparison of latanoprost and dorzolamide in the treatment of patients with open angle glaucoma. AB - BACKGROUND: This study was conducted to compare the effects on intraocular pressure and side effects of monotherapy with either latanoprost or dorzolamide in patients with open angle glaucoma, pseudoexfoliation glaucoma or ocular hypertension. METHODS: Sixty patients with open angle glaucoma or ocular hypertension were recruited to a 3-month study. Previous glaucoma medications were washed out and the patients were randomised to receive either latanoprost 0.005% once daily or dorzolamide 2% three times daily. The follow-up visits were conducted at two weeks, one months and three months of study and intraocular pressures and slit lamp examinations were carried out to look for response of therapy and detect complications. RESULTS: After 3 months, latanoprost reduced mean baseline intraocular pressure from 27.2 +/- 3.0 mm Hg by 8.5 +/- 3.3 mm Hg. The corresponding figures for dorzolamide were 27.2 +/- 3.4 and 5.6 +/- 2.6 mm Hg. The difference of 2.9 mm Hg (95% CI: 2.3-3.6) was highly significant (p < 0.001). Both drugs were well tolerated systemically and locally. CONCLUSION: Latanoprost was superior to dorzolamide in reducing the intraocular pressure, judged from the effect on mean intraocular pressure. The once daily dose in the evening ensures better compliance and the problem of hyperpigmentation of the iris were not encountered. PMID- 15125184 TI - An experience with free scapular flap for reconstruction of lower extremity defects at Aga Khan University Hospital Karachi. AB - BACKGROUND: Reconstruction of large composite tissue defects with exposed vital structures and weight bearing surfaces are extremely difficult for reconstructive surgeons. Coverage of such difficult defects can be achieved with free tissue transfer provided microsurgical expertise is available. This study was carried out to determine the outcome of free scapular flaps performed for lower extremity defects. METHODS: Clinical records of the patients were reviewed and important variables included demographic data, etiology of soft tissue defects, site and size of defect, complications and percentage of wound coverage by flap. Patient satisfaction and objective assessment of flap coverage and motion of involved joints was made. RESULTS: There were 13 male patients with mean age of 30.46 years. The causes of tissue defects were road traffic accident in 8 cases, industrial accidents in 2 and 3 cases had bomb blast, gunshot and gas gangrene one in each case respectively. Nine scapular flaps were performed to reconstruct the defects around the foot, ankle and lower leg; two flaps for amputation stumps and two flaps for defects around the knee and popliteal fossa. Post flap surgery, 3 cases had vascular compromise which required urgent exploration of anastomosis. Two cases were revived and one flap did not survive. With respect to cosmetic appearance, coverage of required defect, performance of activities of daily living and functional range of motion of involved joints, 6 cases were graded excellent and 7 cases were good. CONCLUSION: In our cases of lower extremities defects, scapular flap helped to salvage the limb. In conclusion scapular flap is a versatile flap which can be used for lower extremity defects. PMID- 15125185 TI - Recent trends in laproscopic myomectomy. AB - Recently there is an increasing trend for minimal access surgery (MAS) for treatment of uterine myomas. Laparoscopic myomectomy has provided minimal invasive alternative to laparotomy for subserosa and intramural myomas. It is associated with faster postoperative recovery and potentially less postoperative adhesions. Main concerns are however subsequent fertility, reproductive outcome and long-term recurrence. Other alternatives are laparoscopic assisted myomectomy, laparoscopic ultraminilaparotomic embolised myomectomy, laparoscopically assisted transvaginal myomectomy, myolysis and cryosurgery. Hysteroscopic access is required for sub mucous myomas. The idea of this review is to analyse recent techniques which are used to treat uterine myomas. Recent evidence favours safety and reliability of laparoscopic myomectomy. Prospective randomised controlled trials comparing laparoscopic myomectomy with laparotomy myomectomy will clarify the status further. PMID- 15125186 TI - Mediastinal bronchogenic cyst. AB - A 29 year old woman was found to have a chest mass as part of routine screening with a chest x-ray for an unrelated neurosurgical procedure. Further investigation with a chest CT raised the suspicion of a paravertebral neurogenic tumor. The patient underwent bronchoscopy and resection of the lesion by Video Assisted Thoracic Surgery (VATS). The diagnosis of Bronchogenic Cyst was established after histopathologic examination of the resected tumor. Bronchogenic Cysts are relatively rare primary mediastinal tumors. The paravertebral site is an unusual presentation for these tumors. PMID- 15125187 TI - Reversible obstructive sleep apnea and right heart failure due to massive tonsillar hypertrophy. AB - Obstructive sleep apnea is a relatively common but under diagnosed clinical entity in children. Adenotonsillar hypertrophy is the most common cause for upper airway obstruction in pediatric patients. If the obstruction to upper airways is not relieved, then the child can develop obstructive sleep apnea and its consequences. Treatment is simply to remove the obstruction thereby restoring patency of upper airways. PMID- 15125188 TI - Gastric vascular ectasia. AB - In this paper the condition Gastric Vascular Ectasia (GAVE) is reviewed and two patients of gastric vascular antral ectasia are reported (GAVE). It is suggested that the diagnostic possibility of Gastric antral vascular ectasia should be considered in any elderly patient, more so in females, with persistent unexplained Iron deficiency anaemia. PMID- 15125189 TI - [Biochemical aspects of plant interactions with parasite nematodes. (A review)]. AB - The review summarizes reports on molecular aspects of interactions of phytoparasitic nematodes with plant hosts. Data on the secrets of nematodes affecting plants (elicitors, toxins, products of parasitism genes, etc.) are analyzed and information flow pathways comprising all elements of the plant parasite interaction (from elicitors to defense responses of plant cells), described. Emphasis is placed on the mechanisms whereby plants are protected from nematode invasion (hypesensitivity reactions, apoptosis, phytoalexins, proteinase inhibitors, PR-proteins, etc.). Consideration is given to genetic aspects of plant-parasite relationships. Promising practical approaches to defending plants from phytoparasitic nematodes, developed based on the results of studies of molecular mechanisms of plant-parasite interactions are provided in conclusion. PMID- 15125190 TI - [Inactivation of glucose-6-phosphate dehydrogenase in solution by low- and high frequency ultrasound]. AB - Kinetics of inactivation of glucose-6-phosphate dehydrogenase (G6PDH, EC 1.1.1.49) in 0.1 M phosphate buffer (pH 7.4) within temperature range from 36 to 50 degrees C was studied comparatively under conditions of exposure of enzyme solution to low-frequency (LF, 27 kHz, 60 W/cm2) or high-frequency (HF, 880 kHz, 1.0 W/cm2) ultrasound (USD). Inactivation of G6PDH was characterized by effective first-order rate constants: (k(in)) total (summarized) inactivation; (k(in)*) thermal inactivation; and (k(in)(usd)) ultrasonic inactivation. Dilution of enzyme solution from 20 to 3 nM was accompanied by a significant increase in the values of the three rate constants. The following inequality was valid in all cases: k(in) > k(in)*. The rate constants increased upon increasing the temperature. The Arrhenius plots of the temperature dependencies of k(in) and k(in) (usd) have a salient point at 44 degrees C. The activation energy (Eact) of the total inactivation of G6PDH was higher than Eact for the process of ultrasonic inactivation of this enzyme. The two values were found to depend on USD frequency: Eact in case of inactivation with low-frequency ultrasound (LF USD) was higher than in case of inactivation with high-frequency ultrasound (HF USD). The rate of the ultrasonic induced inactivation of this enzyme substantially decreased in the presence of low concentrations of traps of radicals HO. (dimethylformamide, ethanol, and mannitol). This fact supports the conclusion that free radicals are involved in the mechanism of the G6PDH inactivation in solutions exposed to LF-USD and HF-USD. Ethanol was an effective protector of G6PDH inactivation in enzyme solutions exposed to USD. PMID- 15125191 TI - [Transglycosylation of stevioside by cyclodextrin glucanotransferases of various group of microorganisms]. AB - Cyclodextrin glucanotransferases (CGTases, EC 2.4.1.19), produced by mesophilic, thermophilic, alkaliphilic, and halophilic bacilli, were used for the transglycosylation of stevioside to remove bitterness and aftertaste, with cyclodextrins (CDs) being used as donors. It was shown that CGTases produced by extremophiic microorganisms are effective biocatalysts. Optimal temperature and pH of these enzymes were at pH 6.5-7.5 and 45 degrees C, respectively. The optimal stevioside-to-CD ratio and total concentration of dry matter for the synthesis of best-taste product were 1:1 (w/w) and 11.6%, respectively. PMID- 15125192 TI - [Ecdysone 20-monooxygenase activity of cytochrome P450 in plants and cell culture of Ajuga reptans L]. AB - The concentration of cytochrome P450 and ecdysone 20-monooxygenase activity in plants and callus cell culture of the bugleweed Ajuga reptans L. were determined. The maximal ecdysone 20-monooxygenase activity of cytochrome P450 was found in vegetative rosettes of intact plants. During the stage of flowering, the ecdysone 20-monooxygenase activity of cytochrome P450 in plant leaves was higher than in other organs. It was demonstrated that the content of ecdysteroids in callus cell culture is higher than in the intact plant with concurrent retention of a high ecdysone-20-monooxygenase activity. PMID- 15125193 TI - [Phenyl pyrazolones--novel oxidoreductase redox-mediators for degradation of xenobiotics]. AB - An approach was developed to screening organic compounds for putative activity of redox mediators of oxidoreductases, including laccases and peroxidases, applicable for xenobiotic degradation. The study was carried out with a homogenous laccase preparation from the basidiomycete Trametes hirsuta and horse radish root peroxidase. Compounds belonging to 1-phenyl-3-methylpyrazolones were selected. Spectroscopic and electrochemical investigation of two of the compounds, sodium 1-phenyl-2,3-dimethyl-4-aminopyrazolon 5n(4)-methanesulfonate (PPNa) and 1-(3'-sulfophenyl)-3-methylpyrazolone (SPP), was performed. Electrochemical oxidation of both PPNa and SPP gave rise to high-potential intermediates capable of oxidizing veratryl alcohol; a lignin-modeling compound. Kinetic indices of these compounds were determined in enzymatic reactions with the presence of laccase. It was shown that enzymatic oxidation of SPP by laccase produced high-potential intermediates capable of oxidizing veratryl alcohol to veratric acid. Veratryl alcohol did not oxidize during enzymatic oxidation of SPP by peroxidase. This points to a difference between the mechanisms of enzymatic oxidation of PPNa and SPP by laccase and peroxidase. PMID- 15125194 TI - [Properties and prospects of practical use of extracellular L-glutamate oxidase from Streptomyces sp. Z-11-6]. AB - The properties of extracellular L-glutamate oxidase, isolated and purified from Streptomyces sp. Z-11-6 (specific activity, 50.8 U/mg protein; yield, 40%), were studied. A photometrical method of determination of activities of alanine- and aspartate aminotransferases, based on the use of the L-glutamate oxidase and peroxidase, has been developed. This method is sufficiently sensitive to be used for the determination of aminotransferase activities in biological fluids. The presence of other amino acids did not interfere with the analysis and had no effect on the results of determination. PMID- 15125195 TI - [Quartz sand as an adsorbent for purification of extracellular glucose oxidase from Penicillium funiculosum 46.1]. AB - The procedure of purification of extracellular glucose oxidase (GO, EC 1.1.3.4) from culture-liquid filtrate (CLF) of the fungus Penicillum funiculosum 46.1 using alluvial quartz sand as an adsorbent has been developed. The modification of sand by changing the charge and polarity did not lead to a significant increase in its adsorption capacity towards GO. The effectiveness of sand and aluminum oxide, used as sorbents for isolation of GO from CLF, was compared. Glucose oxidase, isolated from CLF by adsorption on sand, exhibited a greater catalytic activity compared to the enzyme specimens obtained by column chromatography on CLF. Sand adsorbed GO from P. funiculosum 46.1 more effectively than aluminum oxide. It is concluded that sand may be used for fractionation of partly purified GO. PMID- 15125196 TI - [Immobilization of champagne yeasts by inclusion into cryogels of polyvinyl alcohol for preventing cell escape from carrier matrix]. AB - Wine champagnizing, a process involving the use of champagne yeasts immobilized by inclusion into cryogels of polyvinyl alcohol, has been studied. Treatment of yeast cells with the autoregulatory factor d1 was proposed as a means of preventing the cell escape from the carrier matrix. Such a treatment inhibited growth and proliferation processes in yeasts cells, without affecting the activity of fermentation; the resulting champagne had the same organoleptic and chemical characteristics as its counterparts obtained using traditional techniques. PMID- 15125197 TI - [Exoproteinases of the oomycete Phytophthora infestans]. AB - When grown in a medium containing heat-stable potato tuber proteins, the oomycete Phytophthora infestans (Mont.) de Bary produces a set of exoproteinases active at neutral and mildly basic pH values. These extracellular proteinases have been shown by SDS-PAGE with the presence of gelatin to include at least six components differing in molecular weight. Inhibitory analysis and study of the effects of the enzymes on various synthetic substrates show that the culture liquid of P. infestans contains mainly serine proteinases specific to trypsin and subtilisin and metalloproteinases. Their activity is suppressed by proteinase-inhibitor proteins from potato tubers. It is suggested that P. infestans exoproteinases may be the metabolic target for natural proteinase inhibitors from potato. PMID- 15125198 TI - [Dynamics of activity of the key enzymes of polyhydroxyalkanoate metabolism in Ralstonia eutropha]. AB - The dynamics of accumulation of polyhydroxybutyrate (PHB) and the activities of the key enzymes of PHB metabolism (beta-ketothiolase, acetoacetyl-CoA reductase, PHA synthase, D-hydroxybutyrate dehydrogenase, and PHA depolymerase) in the hydrogen bacterium Ralstonia eutropha B5786 were studied under various conditions of carbon nutrition and substrate availability. The highest activities of beta ketothiolase, acetoacetyl-CoA reductase, and PHA synthase were recorded at the stage of acceleration of PHB synthesis. The activities of enzymes catalyzing PHB depolymerization (PHB depolymerase and D-hydroxybutyrate dehydrogenase) were low, being expressed only at stimulated endogenous PHB degradation. The change of carbon source (CO2 or fructose) did not cause any marked changes in the time course of enzyme activity. PMID- 15125199 TI - [Antimicrobial activity of lactic acid bacteria from sour milk products "Narine", "Karine", and "Matsun"]. AB - We studied antimicrobial properties of lactic acid bacteria from sour milk products Narine, Karine, and Matsun. The whey of sour milk products included two major fractions of sugars and L-lactic acid and its sodium and calcium salts. Antimicrobial activity of Narine, Karine, and Matsun was related to the presence of L-lactic acid and its sodium and calcium salts. PMID- 15125200 TI - [Effect of photosynthetic bacteria and compost on degradation of petroleum products in soil]. AB - Addition of diesel fuel and waste engine oil to soil was found to cause biostimulation of hydrocarbon-oxidizing microorganisms. Corynebacteria constitute a large group of hydrocarbon-oxidizing microorganisms. Addition of a liquid culture of photosynthetic bacteria to soil not only facilitates degradation of petroleum products, but also stimulates growth of hydrocarbon-oxidizing microorganisms. Combined addition of photosynthetic bacteria and compost to soil polluted with petroleum products causes even a more significant increase in the count of hydrocarbon-oxidizing bacteria and substantially increases the rate of pollutant degradation. PMID- 15125201 TI - [Germination of basidiospores of Agaricus bisporus]. AB - The type of dormancy and conditions necessary for germination of Agaricus bisporus basidiospores (BS) were studied. BS failed to germinate on starvation agar and required the presence of carbon and nitrogen (asparagine and/or glucose) sources in the medium. Upon 3-week storage, BS germinated after 4-5 days. Heat shock (20 min at 45 degrees C) and the decreased temperature facilitated activation of germination. Heterocyclic compounds stimulating germination of endogenously dormant spores, such as furfural, failed to activate germination. The data obtained suggested an endogenous dormancy of A. bisporus BS differing from zygospores of Mucorales. BS contained 17-19% lipids with a composition of fatty acids differing from those of pileus and stipe of the fruiting body. The soluble carbohydrates of the cytosol amounted to 12% dry spore weight and consisted of mannitol (74%) and trehalose (26%). Unlike BS stored at 2 degrees C, the BS stored for 5 months at 20 degrees C lost their ability to germinate, which correlated with a decrease in the content of trehalose. PMID- 15125202 TI - [Development of magnetic biosorbents and their application in microbial antigen immunoassays]. AB - The feasibility of rendering erythrocytes magnetic and thereby creating magnetic biosorbents (MB Ss) through room temperature exposure to 25-37% iron (II) sulfate solution for 48 +/- 2 h, followed by exposure to 15-35% ammonia water solution for 48 +/- 2 h (with drying after each procedure) was demonstrated. The feasibility of immobilizing ligands on magnetic erythrocytes (ME) and obtaining biological magnetic immunosorbents (BMISs) for further use in EIAs for plague and tularemia antigens was demonstrated. The sensitivity of EIAs involving BMISs amounted to 10 ng/ml and 100 microbial cells per 1 ml. The relative error did not exceed 8%. PMID- 15125203 TI - [Competitive immunochemical determination of antigens using conjugates containing Co(II) and Ni(II)]. AB - A new variant of competitive heterogeneous immunoassay for certain proteinaceous antigens has been developed. The assay is based on the use of the target protein conjugated with Co(II) or Ni(II) ions and immobilized Abs. The effect of catalytic hydrogen release allows quantitation of the metal ion labels by voltammetry at the final step of the assay. The conjugates have been characterized by spectrophotometry, voltammetry, atomic adsorption spectrometry, and nuclear magnetic relaxation. Based on the use of the conjugate RNase diethylenetriaminepentaacetic acid-Co(II) (10:4:4), a competitive immunoassay for RNase has been developed, detecting the target protein in the range 2 x 10(-2)-2 x 10(-4) mg/ml. PMID- 15125204 TI - [Cultivation of Iris ensata Thunb. callus tissue]. AB - A continuous callus culture was obtained from zygotic embryos of Japanese iris (Iris ensata Thunb.) on the Murashige-Skoog medium supplemented with 2 mg/l alpha naphthylacetic acid and 0.5 mg/l 6-benzylaminopurine (BAP). It was found that a successful callusogenesis required isolated embryos at the wax stage of endosperm development. The optimal combination of phytohormones for the growth of callus tissue was 1 mg/l 2,4-dichlorophenoxyacetic acid and 0.5 mg/l BAP. The pigment composition of I. ensata callus tissue was studied. It was demonstrated that subcultivated callus tissue contained red pigments of flavonoid nature. Under stress cultivation conditions, yellow pigments were formed and the content of red pigments increased. PMID- 15125205 TI - [Effect of systemic signaling molecules on the rate of spread of the immunizing effect of elicitors over potato tissues]. AB - Mobile systemic signaling molecules (salicylic and jasmonic acids) enhance and accelerate the spread of systemic immunizing effect of elicitors (arachidonic acid and chitosan) over potato tuber tissues (Solanum tuberosum L.). PMID- 15125206 TI - [Highest level of division in classification of organisms. 2. Archaebacteria, eubacteria and eukaryotes]. AB - In three-domain system of organic world archaebacteria are considered as the third form of life alongside with eubacteria and eukaryotes. The author gives brief characteristics of all three groups with special focus on such diagnostic attributes as: plasmatic membrane and cellular wall, flagella, protein transcription, replication, topoisomerases, transcription, translation, glycosylation, chaperons and chaperonins, proteasomes and exosomes, histones, ATP ases. The three-domain system has been proposed by several scientists but principal ideas were put by C. Woese. The systematics according Woese should reflect contemporary level of our knowledge of organisms. In the historical plan it once had to refuse dividing the organic world into plants and animals but accept the division into prokaryotes and eukaryotes. The science however goes further and turns now to the new level of generalizations based on the molecular aspects of cellular structures and processes. From this point of view, both plants and animals are uniform. As to prokaryotes they appeared to be non monolithic group because of essentially different transcriptional and translational mechanisms. Therefore the detachment of archaebacteria as an independent group was the important step in the development of systematics. At the same time the three-domain system of organisms is typological and requires correction according to data on phylogenetic relatedness of these groups. PMID- 15125207 TI - [Epigenetic and synergistic types of inheritance of the reproductive characters in angiosperms]. AB - The author considers three types of hereditary memory (structural, cell and signal), that are realized on different levels of biological organization. These three types of hereditary memory correspond to three types of reproduction: self replication, cell division and reproduction s. str. Reproductive characters are exemplified with three essential characters in angiosperm plants: dimorphism in population by flower sex; mono-, di- and trystyly of flowers; uni- and biparental mode of seed reproduction. All these characters are considered as "supercharacters" that are controlled by gene ensembles. The correspondence between three types of reproduction and three types of hereditary memory are discussed. The authors reviews also the role of polyploidy (auto- and endoploidy) in the inheritance of reproductive condition. From the information theory point the increase in cell ploidy causes the growth of uncertainty in expression of genes and gene ensembles thus creating new type of variability--epigenetic variation. The change of reproductive strategy in plants is regulated by state of gene and gene ensembles and does not demand structural changes in genome. The reproductive characters of plants in spite its complex structure are inherited in number of generations as a discrete Mendel characters by mono-, di-, ot trihybrid schemas. PMID- 15125208 TI - [Social learning in animals: comparative analysis of different forms and levels]. AB - Social learning as one of the key concepts of cognitive ecology includes different forms of behavioural displays from relatively simple, such as "social release" and "stimulus enhancement" up to "teaching" and "cultural transmission" in animal communities. Rapid development of this fields resulted in some contradictions in methods and terminology. In this review different forms and levels of social learning are analyzed. Ecological aspects of social learning are connected with diet shaping, fear of predators and mate choice. The first aspect is the most studied but still discussible. Social learning being an intricate component of feeding behaviour matches with innate behaviour, imprinting as well as early associative learning. Investigation of cognitive aspects of social learning going back to Thorndike's crucial question "Do apes ape?" are now developing into series of questions including even: "Do ants ape?". Elaboration of universal methods of comparative studying of social learning such as "artificial fruit" and "two ways/one outcome" has essentially enlightened these questions and made comparative analysis possible. Large continuum of displays of cognitive skills in social learning has been revealed in non-primate species. One of the discussible issues in the role of social learning is distribution of innovations. Many authors have investigated this intriguing aspect of animal behaviour in different ways, such as long field observations as well as laboratory experiments based on "artificial innovators" that is specimens specially taught by experimentalists. Many impressive results were obtained; in particular it turned out in contradiction with some mathematical models that individuals in groups are rather different in their psychophysiological predisposition to innovative behaviour. Role of teaching in such different forms of behaviour as shaping of species-specific behavioural patterns and spread of innovations is considered. Although the majority of animals in wild populations are not good teachers and pupils, some cultural aspects of behaviour were recently revealed, mostly in primates. At the same time some classical results concerning cultural transmission of new patterns (for example, bottle-opening in tits) were experimentally revised. Many problems still remain unsolved, in particular, how spread of innovations may favour prosperity of populations; to what degree behavioural peculiarities of local groups may be determined by processes of social learning; which internal and external factors and under what circumstances invest into social learning in natural environment. PMID- 15125209 TI - [Symbiotic relationships between ants and aphids]. AB - The classical and contemporary literature devoted to symbiotic relationships between ants and aphids was analyzed. The most studied issues are: the totality of morphological and anatomical adaptations of partners to symbiosis, different degrees of aphids myrmecophilia, benefits and costs of insects during interaction. A lot of issues are still discussible. For instance, it is unknown whether the trophobiotical relationships between ants and aphids are mutualistic or intermediate between mutualism and exploitation. To clarify this vague point the study of symbionts behaviour is of great importance, however, it remains practically non-investigated either for aphids or ants. It is known only that nonmyrmecophilous aphids living within galls have rather complicated behaviour- their colonies are guarded with specialized castes of aphids-soldiers. As for the ants, till nowadays trophobionts (individuals looking after the aphids), which make up special functional group within ants family, have been described as passive foragers being occupied only with the honeydew collection and transportation. The recent investigations revealed that ants use behavioural schemes with different degree of complication interacting with aphids: from lonely foraging to professional specialization in working groups. PMID- 15125210 TI - [The meaning of vegetation classification for modern ecology]. AB - Eco-floristic classification (Brawn-Blanquet system) is useful since in similar manner it can establish ecological units of different level (syntaxons) that have high indicative value. Syntaxons can be used as markers of terrestrial ecosystems of different range. Although this system was proposed by analogy with plant taxonomy it reflects vegetation continuum. The application of this approach is based on certain standards and supported by developed software. The elements of arbitrariness in Brawn-Blanquet method are constrained by rules of strict recommendations. The method is considered as necessary in studies of biodiversity in protected areas. PMID- 15125211 TI - [The spatial structure of lichen Hypogymnia physodes (L.) Nyl]. AB - Structure analysis based on regular morphological variability at organism level is applied to study the shape and spatial construction of lichen Hypogymnia phyzodes (L.) Nyl. Some structural were determined at juvenile stage of development with the use of this approach. Five of them are dominant. Besides, five the most frequent encounter types of thallus branching were described. It is shown that thallus structure at adult stage is usually built from two main elements forming axial dichotomy. Analysis of thallus shape indicates that transformation of dichotomy elements into trichotomy ones may also take place during growth and development of thallus. PMID- 15125212 TI - [Some notes on relation between taxon and character in taxonomy (regarding the paper of A.A. Stekol'nikov "A problem of truth...", Zhurnal Obshchei Biologii. 2003.V.64. No 4. P.367-368)]. AB - Under brief consideration is the problem of primary or secondary status of the judgments about taxa relative to the judgments about characters in the biological classifications. The following formal definition of taxonomic system (classification) TS is provided: TS = BT[T, C(t), R(t), R(c), R(tc)], where BT is a biological theory constituting content-wise background of the system, T is a set of taxa, C(t) is a set of taxonomic characters, R(t) is a set of relationships among taxa (similarity, kinship, etc.), R(c) is a set of relationships among characters (homology, etc.), and R(tc) is a set of correspondences among taxa and characters. The latter correspondences may be complete or incomplete. At ontological level, there two basical traditions exist in biological systematics regarding R(tc) according to which the biological diversity is patterned either as a set of groups of organisms (taxa) or as a set of their properties (characters). In the first case, taxon is "primary" relative to character (in cladistics); in its opposite, character is "primary" relative to taxon (in scholasticism, classical typology, classical phylogenetics). At epistemological level, incompleteness of the taxon-character correspondence makes classificatory procedure iterative and taxonomic diagnoses context-dependent. The interative nature of classificatory procedure makes the "primary" or "secondary" status of both taxa and characters relative and alternating. This makes it necessary to introduces a kind of uncertainty relation in biological systematics which means impossibility of simultaneous definition of both extensional and intentional parameters of the taxonomic system at each step of classificatory iterations. PMID- 15125213 TI - Comparative study of the effectiveness of thyroxine and steroids on reduction of neonatal morbidity: outcome at 20 months follow-up. AB - This study analyzes health, growth and neurodevelopmental outcome of infants who received prenatal corticosteroids with or without thyroxine for fetal lung maturation. During a 12 month period infants from a prospective double blind study who received either steroids or steroids combined with thyroxine for pulmonary maturation and who had reached 18 months of age were recalled for evaluation of health status, growth parameters and neurodevelopmental outcome using the Bayley Scales 2nd edition (BSID-II). Mental developmental index (MDI), psychomotor developmental index (PDI), language developmental age (LDA), cognitive developmental age (CDA), and behavioral rating scales (BRS) were compared for the two treatment groups. The Hollingshead Socioeconomic Status Index was determined for each infant. Of a total of 134 patients enrolled during the study period, 66 patients return for follow up. Data from 60 patients was included in the final analysis. Of these, 32 had received the combination regimen and 28 had solely received only steroids. Demographics and neonatal morbidity were similar in both groups. No statistical differences in growth parameters, hospital admissions, respiratory problems, surgical procedures or frequency of infections were found. Neurodevelopmental parameters (MDI, PDI) were similar in both treatment groups, although, below normal in both groups. Language delay was more common in infants who received prenatal corticosteroids. There were no difference in the incidence of neurologic abnormalities. The addition of thyroxine to steroids did not affect growth or neuro-developmental outcome of the infants at 18 to 22 postnatal age. PMID- 15125214 TI - Hereditary and acquired angioedema: experience with patients in Puerto Rico. AB - Hereditary (HAE) and acquired (AAE) angioedema are vascular reactions involving the sub mucosal tissues, representing localized edema caused by dilatation and increased permeability of the capillaries. HAE and AAE are clinical disorders characterized by angioedema that require prompt differentiation from other causes of angioedema in order to receive the most pertinent and effective therapeutic interventions. The aim of this report is to describe the clinical characteristics of patients with both HAE and AAE identified and followed at the Immunology Clinic of the University Hospital at the Puerto Rico Medical Center, their response and side effects to danazol therapy and their comparison with other series of similar patients reported in the literature. Overall, the patients in this sample presented a similar clinical profile compared to other reported series in the literature. PMID- 15125215 TI - Potentially serious infections in the aging person: diagnosis, treatment and prevention. AB - Infections in the elderly patient are a challenge, since the classical signs of infection are absent or ill defined. The present paper describes the presentation, diagnosis, clinical manifestations and treatment for a selected group of potential serious infections including influenza, bacterial pneumonia, urinary tract infections as well as infections caused by multiresistant bacteria, like vacomycin-resistant enterococcus and methicillin resistant S. aureus. We conclude with the need for prevention in the older person with the use of vaccines, specifically the influenza and pneumococcal vaccine as well as the prevention of urinary infections. Influenza is a significant cause of morbidity, whose ill effects can be prevented in many older persons with the use of a vaccine. The use in prophylaxis and treatment of antiviral agents like amantadine, rimatadine, and oseltamivir is presented. Bacterial pneumonia is one of the leading causes of death in the USA among the older persons. The emergence of drug resistant Streptococcus pneumoniae leads to the consideration as empiric therapy the newer fluoroquinolones or the use of third or fourth generation cephalosporis. Of importance is the use of pneumococcal vaccine among people age 60 or above. The frequency of urinary tract infections among the elderly is of primary although in many instances important do not require treatment. When infection of the urinary tract is diagnosed, most authors use a fluoroquinolone as empiric theraphy. The emergence of multiresistant bacteria like methicillin resistant S. aureus and or vancomycin resistant enterococci leads to the need to consider new agents like quinipristin-dalfopristin, linezolid and deptomycin in the management of such patients. PMID- 15125216 TI - Antibiotics in older adults. AB - Antibiotics are frequently prescribed in the older person, the dosification needs special care, since the pharmacokinetic parameters changes with aging and the side effects can be different in the older person. The creatinine clearance changes and we must modify the way we prescribe such antibiotics to the elderly, calculating. The variety of antibiotics now available led us to consider this paper in which we have presented the antimicrobial agents that can be considered in the treatment of the older person. We present several groups: the penicillins, cephalosporins, monobactams, carbapenems and betalactamase inhibitors or the great betalactam group. Other trimetroprin-sulfame-thoxazole, the newer macrolides (azithromycin and clarithromycin) as well as the aminoglycosides, vancomycin, clindamycin, metroridazole. The indications and contraindications are presented and reviewed. PMID- 15125217 TI - Status of cardiovascular disease in Puerto Rico. AB - For the last 40 years diseases of the heart has ranked as the primary cause of death in Puerto Rico, being cancer the second. In the National Vital Statistics of the Department of Health of Puerto Rico for the year 2002, cancer is listed as the main cause of death and diseases of the heart ranks as the second most frequent. This unexpected change is mainly due a new classification by ICD-10, that now substitutes ICD-9, in association to a change introduced by using the 2000 population census rather than the 1990 census used until 1999. Additional relevant information concerning life expectancy at birth, death by specific causes in the last 5 years and statistics about invasive surgical and medical services rendered to cardiovascular patients in the year 2003 are included. A summary of the situation concerning postgraduate training programs in Puerto Rico in different cardiovascular subspecialties is also included. PMID- 15125218 TI - The introduced free-ranging rhesus and patas monkey populations of southwestern Puerto Rico. AB - Rhesus (Macaca mulatta) and patas (Erythrocebus patas) monkeys escaped to the mainland of southwestern Puerto Rico (SWPR) from research colonies on small offshore islands during the 1960s and through 1982. A three year study (1990 1993) combined radio-telemetry with visual observations to collect information on population sizes, the composition of social groups, their daily movements, and their home ranges. Two populations of rhesus monkeys were identified in SWPR: one within the study area in Sierra Bermeja and a second population located 10 km north of the study area. The size of the Sierra Bermeja rhesus population was derived from escapees from research colonies and at the time of the study was 65 85 individuals. Within their home range area (3.7 km2) the density of this population was >>18.9 individuals/km2. A second rhesus population was found in a mountainous region 10 km north of the study area. This population consisted of one (or two) heterosexual groups with a total of 40-45 individuals. Although a primary characteristic of this species in India is its ability to live as a commensal with humans, the rhesus monkey populations of SWPR are extremely shy and elusive, they avoid contact with humans. The patas monkey population consisted of >>120 individuals in four heterosexual groups and several all-male bands. There was no evidence of patas monkeys outside the study area. Within their home ranges (26.8 km2) the population density was 4.47 individuals/km2). Patas monkeys have not previously been considered a territorial species, their behavior in SWPR suggested territoriality. In contrast to studies in Africa, where the amount of home range overlap between patas monkey groups in high, in SWPR the amount of range overlap between groups is small and each group uses areas with clearly defined boundaries. PMID- 15125219 TI - Allergy to house dust mites and asthma. AB - House dust mites have been shown to be important sources of indoor allergens associated with asthma and other allergic conditions. Asthma is a chronic respiratory disease that affects millions of people worldwide, and numerous scientific studies have shown that the prevalence of asthma is increasing. The most common dust mite species around the world include Dermatophagoides pteronyssinus (Dp), Dermatophagoides farinae (Df), Euroglyphus maynei (Em) and Blomia tropicalis (Bt). Over the past three decades, many important allergens from these species have been identified and characterized at the molecular level. The biological function of several house dust mite allergens has been elucidated, with many of them showing enzymatic activity. However, Bt allergens remain the least studied, even though this mite is very common in tropical and subtropical regions of the world, including Puerto Rico. Therefore, it is very important to include Bt in diagnostic and therapeutic strategies for house dust mite induced allergy and asthma, particularly in areas where Bt exposure and sensitization is high. Recombinant DNA technology, as well as other molecular biology and immunological techniques, have played a fundamental role in advances towards a better understanding of the biology of house dust mites and their role in allergic diseases. This kind of study also contributes to the understanding of the complex immunologic mechanisms involved in allergic reactions. The development of effective diagnostic and therapeutic approaches depends on the continuity of research of house dust mite allergens. The objectives of this review are to describe the most important aspects of house dust mite allergy and to acquaint the scientific community with the latest findings pertaining to house dust mite allergens, particularly those derived from Bt. PMID- 15125220 TI - Extracorporeal membrane oxygenation: successful bridge to pediatric heart transplantation. AB - Since the year 2000, extracorporeal membrane oxygenation (ECMO) support for cardiac failure has been employed in the post operative care of children at the Centro Cardiovascular de Puerto Rico y el Caribe. Our experience with the application of ECMO had been limited to circulatory support after repair of congenital cardiac lesions. We report the first case in Puerto Rico where ECMO was used successfully as bridge to pediatric heart transplantation. PMID- 15125221 TI - Pulmonary hypoplasia in Jarcho-Levin syndrome. AB - Jarcho-Levin syndrome, also known as spondylothoracic dysplasia and characterized by short trunk dwarfism, "crab-like" rib cage, with ribs and vertebral defects; it is not uncommon in Puerto Ricans. Many patients die in early infancy due to respiratory compromise associated to lung restriction and the reported cases emphasize mostly the skeletal malformations associated to the syndrome. We report the autopsy findings in a newborn with isolated Jarcho-Levin syndrome emphasizing pulmonary pathology. He was a pre-term male who died of respiratory failure at three hours old and, autopsy findings confirmed the clinical diagnosis. Internal examination showed hypoplastic lungs with normal lobation. The histological structure appeared normal and relatively mature; the diaphragm showed eventration and unilateral absence of musculature. This case shows the worst spectum of the Jarcho-Levin syndrome: pulmonary hypoplasia not compatible with extrauterine life. Since thoracic restriction is present during the fetal period, the degree of pulmonary hypoplasia probably defines survival beyond the neonatal period. PMID- 15125222 TI - [From identification of genomic polymorphism to diagnostic and prognostic markers of human epithelial tumors]. AB - The review considers the results obtained by several groups in the fields of identification of polymorphic loci in the human genome, localization and analysis of genes associated with epithelial tumors of various origins, and generation of molecular markers of socially important oncological diseases. In the first two cases, work was initiated and supported by the Russian program Human Genome. To find new polymorphic loci in the human genome, di-, tri-, and tetranucleotide repeats were searched for in an ordered cosmid library of chromosome 13, NotI and cosmid clones of chromosome 3, and in brain EST. In total, nine polymorphisms and almost 200 STS were identified. Markers of NotI clones of chromosome 3 were associated with particular genes. Polymorphic loci NL1-024, NL2-007, and EST04896 were employed in analysis of deletions from chromosome 3p in tumor DNA. Deletion mapping of 3p in epithelial tumors of five types revealed six critical regions containing potential tumor suppressor genes. Of these, two were in the distal region of chromosome 3p and four, in region 3p21.3. A significant correlation was observed for the frequency of allelic deletions and the stage and the grade of tumors (P < 0.05). On the strength of these findings, genes of region 3p were associated with both tumor development and progression, and proposed as prognostic markers. Regions LUCA and AP20 (3p21.3) showed a high (90%) frequency of aberrations, including homozygous deletions in almost 20% cases. The peak of allelic deletions from region D3S2409-D3S3667 (600 kb) was statistically valid (P = 10(-3)). Regions AP20 and D3S2409-D3S3667 (3p21.3) were for the first time associated with tumorigenesis. Clusters of tumor suppressor genes were identified in regions LUCA, AP20, and D3S2409-D3S3667. Methylation of RASSF1A and RARbeta2 (3p) was associated with early carcinogenesis, and that of SEMA3B, with tumor progression. These findings are useful for early diagnostics and post-surgery prognosis of tumors. PMID- 15125223 TI - [Molecular markers of various stages of nonsmall cell lung cancer development]. AB - Studies performed in the recent decade in the Laboratory of the Regulation of Cell and Virus Oncogenes associated structural and functional defects of several oncogenes and tumor suppressor genes with various stages of non-small-cell lung cancer. High risk of lung cancer was established for carriers of rare alleles of the Hras1 minisatellite, the hypermethylated p16INK4A promoter, and microsatellite defects in chromosome regions 3p12, 3p14.2, 3p22-24, 3p21, 3p25, 9p21, and 17p13. Analysis of the Hras1 minisatellite and microsatellites located in two minimal deletion overlap regions of 1p36 was shown to allow a more reliable prognosis in lung cancer. The results testified again that panels of molecular markers are useful for individual risk assessment, early and differential diagnosis, and prognosis in cancer. PMID- 15125224 TI - [Structural-functional characteristics of the 13q14 region of the human genome in the search for potential tumor suppressor genes]. AB - Works on chromosome 13 mapping supported by the Russian program Human Genome are reviewed. Emphasis is placed on studies of region 13q14.3, which is often lost in some human tumors and potentially contains tumor suppressor genes (TSG). A strategy of TSG search is described. As the resolution of genome analysis improved, a minimal overlap of genetic loss in B-cell chronic lymphocytic leukemia (B-CLL) was established for chromosome 13. A map of expressed sequences was constructed for the region containing the overlap, and candidate TSG of chromosome 13q14 were identified. The candidate genes were analyzed both structurally and functionally, and their possible role in tumorigenesis was considered. Assuming haploinsufficiency as a genetic mechanism controlling B-CLL, a new strategy was proposed for mutation screening aimed at identifying potential TSG of region 13q14. PMID- 15125225 TI - [Diagnostics for epigenetic pathology in hereditary and oncologic diseases]. AB - The review considers the epigenetic defects and their diagnostics in several hereditary disorders and tumors. Aberrant methylation of the promoter or regulatory region of a gene results in its functional inactivation, which is phenotypically similar to structural deletion. Screening tests were developed for Prader-Willi, Angelman, Wiedemann-Beckwith, and Martin-Bell syndromes and mental retardation FRAXE. The tests are based on allele methylation analysis by methylation-specific or methylation-sensitive PCR. Carcinogenesis-associated genes (RB1, CDKN2A, ARF14, HIC1, CDI, etc.) are often methylated in tumors. Tumors differ in methylation frequencies, allowing differential diagnostics. Aberrant methylation of tumor suppressor genes occurs in early carcinogenesis, and its detection may be employed in presymptomatic diagnostics of tumors. PMID- 15125226 TI - [Interaction of viral and cellular genes in cervical tumors]. AB - The results obtained in the Laboratory of Molecular Biology of Viruses, CRC carried out in the framework of the Human Genome program and devoted to the study of the activity of cell and viral genes in cervical cancer are summarized. DNA of human papillomaviruses persists in tumors both in episomal and integrative forms. Integration may occur in different regions of chromosomes. Viral transforming genes E6 and E7 are always present in tumor cells, while antibodies to these proteins are detected only in approximately 30% of patients. Loss of heterozygosity is detected on long and short arms of chromosome 6; some such cases are manifest already at the early stages of tumor progression, while others are typical of the late stages. Several genes that are potentially involved in tumorigenesis and are subject to hypermethylation in CpG islands were identified. Methylation of several genes is observed in approximately 30% of tumors. Tumor progression is associated with increased expression of p16ink4a, an inhibitor of cyclin-dependent kinases. PMID- 15125227 TI - [Serological study of a repertoire of human cancer antigens and autoantigens]. AB - The spectrum of human antigens allows a monitoring of various pathological processes such as autoimmune disorders and tumorigenesis. Serological analysis of cDNA expression libraries (SEREX) is now used to search for new cancer-associated antigens, which are potential diagnostic markers or targets for immunotherapy of cancer. The results obtained for several solid tumors are reviewed. Groups of antigens detectable by SEREX, causes of immunogenicity of autoantigens, and prospective implication of antigens in diagnostics and monitoring of cancer are discussed. PMID- 15125228 TI - [Insertional polymorphism of the CYP2E1 gene in infiltrative pulmonary tuberculosis in populations of Bashkortostan Republic]. AB - Insertion polymorphism Ins96 of the CYP2E1 promoter region was for the first time studied in three ethnic groups of Bashkortostan. Population-specific features of genotype and allele frequency distributions were observed. The CYP2E1 polymorphism was associated with infiltrative pulmonary tuberculosis in the Bashkortostan population. PMID- 15125229 TI - [Search for the association of polymorphic markers for genes coding for antioxidant defense enzymes, with development of diabetic polyneuropathies in patients with type 1 diabetes mellitus]. AB - The allele and genotype frequency distributions of polymorphic markers of genes coding for antioxidant enzymes were compared for type 1 diabetes mellitus patients with or without diabetic polyneuropathy (DPN). The groups (total 180 patients) had nonoverlapping (polar) phenotypes. Group DPN+ included 86 patients with DPN and diabetic record no more than 5 years. Control group DPN- included patients without DPN and diabetic record of at least 10 years. Comparative analysis with Fisher's exact test revealed a significant difference in allele and genotype frequency distributions of the T(-262)C polymorphic marker of the CAT gene. Polymorphic markers C1167T of the CAT gene, Pro/Leu of the GPX1 gene, 0/+ of the GSTT1 gene, and 0/+ of the GSTM1 gene showed no significant difference in allele or genotype frequency distribution. On this evidence, these markers were not associated with DPN in the sample examined. PMID- 15125230 TI - [Creation of a new construct for cloning DNA and modeling the structure of Drosophila polytene chromosomes]. AB - Modification of P element-based transformation vector pCaSpeR3New yielded a new construct, pICon, which contains the structural region of the Escherichia coli lacZ, the adjacent 5' and 3' regulatory regions of hsp70, pUC19, and two tandem FRTs. Owing to the hsp70 promoter, the pICon insertion site may be localized on polytene chromosomes after heat shock by light or electron microscopy. The pUC19 sequence with a polylinker allows cloning of the genomic sequence adjacent to the 3' end of pICon by the rescue of the P-element target. Functional FRTs allow insertion or deletion of various DNA fragments. The construct is large (22046 bp), forms easily detectable structures in polytene chromosomes, and may be used to study the structural and functional organization of the Drosophila melanogaster genome, in particular, to elucidate the causes of banding pattern formation. To map the molecular boundaries of interband 3C6/C7, the DNA sequence of this region was cloned between the two FRTs. PMID- 15125231 TI - [Bank of samples from the Prof_Pat protein family, assessment of efficacy]. AB - The PROF_PAT protein pattern database has been created and maintained so as to comprise the maximal number of the SWISS-PROT + TrEMBL proteins as patterns. The present paper describes some characteristic features of PROF_PAT to assist the potential user. New amino acid sequences (10938) from the SWISS-PROT database have been analyzed to determine the boundary values of the "score" parameter to distinguish random and significant similarities. Analysis through the Internet of 20 amino acid sequences having no descriptions in the TrEMBL database demonstrated that PROF_PAT, being highly competitive with its counterparts in specificity, surpasses them in amplitude and variety of proteins, working several times as fast. The real representation of protein families in the PROF_PAT database (release 1.11), which contains 50,149 patterns of 344,429 proteins, has been estimated at 31,450. PMID- 15125232 TI - [Analysis of expression of a series of lymphoma-specific genes in human fibroblasts immortalized by SV40 virus]. AB - Subtraction hybridization was earlier used to obtain cDNA clones corresponding to human genes upregulated in HIV-associated centroblast lymphoma (CL) as compared with HIV-associated immunoblast lymphoma (IL). With inverse subtraction hybridization, clones were isolated that correspond to genes upregulated in IL compared with CL. In addition to cDNAs characterized earlier, the resulting clones contained several (seven CL-specific and three IL-specific) sequences with unknown functions. To identify the lymphoma-specific genes that are overexpressed in early carcinogenesis, Northern blotting was used to assess the level of gene transcription in two human fibroblast lines and in their derivatives immortalized with either a temperature-sensitive mutant of SV-40 or with pSV3neo carrying the SV-40 A gene, considering the latter as a model of early cell malignant transformation. Increased expression in at least one immortalized line compared with normal fibroblasts was observed for set, a-myb, ND1, ND2, ND4 (NADH dehydrogenase subunits 1, 2, and 4), COX2, COX3 (cytochrome-c-oxidase subunits 2 and 3), KIAA0129, and the gene corresponding to cDNA hss2-1-7-10. High expression of these genes was assumed to be associated not only with lymphomogenesis, but also with early transformation (immortalization) of other, nonlymphoid cells. Expression of the calpain gene and the gene corresponding to cDNA hss2-2-9-5 proved to be lower in immortalized than in normal fibroblasts. This was considered indicative of an alternative mechanism of fibroblast transformation or of different processes regulating the expression of these genes in early and late carcinogenesis. PMID- 15125233 TI - [RNA-interference, induced by transient and continuous expression of hairpin RNA in cells from Drosophila and mammals]. AB - RNA interference (RNAi) may be induced by a plasmid with an inverted repeat (IR) sequence directing transcription of hairpin-type double-stranded RNA (dsRNA). This study examines the effects of changing various parameters of IR constructs on Drosophila and mammalian RNAi, using the dual luciferase system, RNAi activity was found to vary depending on IR length ass well as the length and sequence of the internal loop separating sense and antisense sequences. Both transient and stable RNAi occurred in Drosophila cultured cells. Although transient DNA mediated RNAi was noted in most mammalian cells, no mammalian cells stably possessing IR sequences and hence RNAi activity could be obtained. In Drosophila, DNA-mediated RNAi was considerably weaker than long-dsRNA-mediated RNAi. The cytological data indicated that this was most probably caused by abortive processing of hairpin RNA produced within cells. DNA-mediated RNAi was examined at the level of Drosophila individuals using extramacrochaetae as a model gene, and the presence of an intron sequence in the single-stranded loop region was shown to be essential for effective RNAi. PMID- 15125234 TI - [Interaction of effect on secretion of alkaline phosphatase from E. coli by charge of the N-terminal part of the signal peptide and proteins SecB and SecA]. AB - The cytoplasmic step of posttranslational secretion in Escherichia coli is catalyzed by export-specific chaperone SecB and translocational ATPase SecA. In addition, the efficiency of secretion depends on the charge of the signal peptide (SP). Substitution of positively charged Lys(-20) with noncharged Ala or negatively charged Glu in the N-terminal region of SP of the alkaline phosphatase (PhoA) precursor (prePhoA) was shown to decrease the PhoA secretion in the periplasm. The effect on secretion increased in the absence of SecB and was especially high on SecA inactivation. A change in SP charge strengthened the SecA and SecB dependences of secretion. On evidence of immunoprecipitation, the charge of the N-terminal region of SP had no effect on prePhoA interaction with the cytoplasmic secretion factors, suggesting no direct binding between this region and SecA or SecB. Yet the charge of the N-terminal region proved to affect the functions of SP as an intramolecular chaperone and a factor of prePhoA targeting to the membrane in cooperation with SecA and SecB. PMID- 15125235 TI - [Structural organization and control of expression of the sop-operon of linear plasmid prophage N15]. AB - Stable inheritance of bacterial chromosomes and low-copy-number plasmids depends on the active partition of replicated molecules between daughter cells. The partition mechanism is well known for circular plasmids F and P1. The mechanism of partition of linear replicons was studied with the example of bacteriophage N15, which persists as a linear plasmid with covalently closed ends on lysogeny, rather than integrating into the Escherichia coli chromosome. Since stable inheritance of N15 is due to the sop operon homologous to sop of the F plasmid, the control of expression of the N15 sop genes was analyzed. The sop promoter (Psop) contains a binding site for bacterial IHF and five CTTTGC copies, which overlap the -35 and -10 elements. The Sop proteins were shown to interact with a Psop-containing DNA fragment in vitro. Transcription of the sop operon is regulated by the Sop proteins: SopA represses Psop, and SopB enhances the repression, having no effect on the promoter activity in the absence of SopA. In N15 lysogenic cells, Psop proved to be repressed. This regulatory mechanism was assumed to ensure production of SopA and SopB in amounts required for the segregation stability of N15 and to neutralize occasional fluctuations of their concentration in the cell. PMID- 15125236 TI - [Molecular morphology of eukaryotic class I translation termination factor eRF1 in solution]. AB - The integral structural parameters and the shape of the molecule of human translation termination factor eRF1 were determined from the small-angle X-ray scattering in solution. The molecular shapes were found by bead modeling with nonlinear minimization of the root-mean-square deviation of the calculated from the experimental scattering curve. Comparisons of the small-angle scattering curves computed for atomic-resolution structures of eRF1 with the experimental data on scattering from solution testified that the crystal and the solution conformations are close. In the ribosome, the distance between the eRF1 motifs GGQ and NIKS must be shorter than in crystal or solution (75 versus 107-112 A). Therefore, like its bacterial counterpart RF2, the eukaryotic eRF1 must change its conformation as it binds to the ribosome. The conformational mobility of eukaryotic and prokaryotic class-1 release factors is another feature making them functionally akin to tRNA. PMID- 15125237 TI - [Mutual effect of translation initiation factors in binding with IRES-element of encephalmyocarditis virus RNA]. PMID- 15125238 TI - [Model of aggregation of pigments in the chlorosomal antenna of the green bacteria Chloroflexus aurantiacus]. AB - Independent experimental and theoretical evaluation was performed for the adequacy of our previously proposed general molecular model of structural organization of light-harvesting pigments in chlorosomal bacteriochlorophyll (BChl) c/d/e-containing superantenna of different green bacteria. Simultaneous measurement of hole burning in the optical spectra of chlorosomal BChl c and temperature dependence of steady-state fluorescence spectra of BChl c was accomplished in intact cells of photosynthetic green bacterium Chloroflexus aurantiacus; this allows unambiguous determination of the structure of exciton levels of BChl c oligomers in this natural antenna, which is a fundamental criterion for adequacy of any molecular model for in vivo aggregation of antenna pigments. Experimental data were shown to confirm our model of organization of oligometric pigments in chlorosomal BChl c antenna of green bacterium Chloroflexus aurantiacus. This model, which is based on experimental data and our theory of spectroscopy of oligomeric pigments, implies that the unit building block of BChl c antenna is a cylindrical assembly containing six excitonically coupled linear pigment chains whose exciton structure with intense upper levels provides for the optimal spectral properties of the light-harvesting antenna. PMID- 15125239 TI - [Conformational status of apomyoglobin in the presence of phospholipid vesicles at neutral pH]. AB - The conformational state of sperm whale apomyoglobin (apoMb) was studied at neutral pH in the presence of negatively charged vesicles using near- and far-UV circular dichroism, tryptophan fluorescence, differential scanning microcalorimetry, and fast performance liquid chromatography. Under these conditions, the apoMb structure undergoes transition from its native to an intermediate state. In this state the protein loses its rigid native structure but retains its secondary structure. However, the environment of tryptophan residues remains rather hydrophobic. This intermediate state of apoMb shows properties similar to those of its molten globule state in solution. It is shown that apoMb can bind to negatively charged phospholipid vesicles even at neutral pH. A possible functional role of this intermediate state is discussed. PMID- 15125240 TI - [Effect of tumor necrosis factor DNA on immune response to DNA immunization against herpes simplex virus]. AB - A study was made of the adjuvant effect of the mouse tumor necrosis factor alpha (mTNF alpha) on DNA immunization against the herpes simplex virus type 1 (HSV1). The HSV1 gD gene (pDNAgD) served as an immunogen; mTNF alpha or its gene cloned in an eukaryotic expression vector (pDNAmTNF) were used to modulate the immune response. Double immunization with pDNAgD led to a sixfold increase in the in vitro T-cell response, a high (1:2000) titer of anti-HSV1 antibodies (including virus-neutralizing antibodies), an increase in IgG2a/IgG1 (suggesting a shift of the immune response to the Th1 type), and no change in CD4/CD8 T-cell ratio. A single injection of mTNF alpha along with inactivated HSV1 allowed a twice higher antibody titer and a fourfold higher T-cell response as compared with immunization with HSV1 alone. Double immunization with both pDNAgD and pDNAmTNF increased the titer of anti-HSV1 antibodies and the T-cell response by factors of 8 and 1.5, respectively, as compared with immunization with pDNAgD alone. However, the protective effect was significantly lower with the two plasmids than with pDNAgD (73 vs. 100%). Thus, DNA immunization with pDNAgD induced both B- and T-cell responses and completely protected mice from a lethal doze of HSV1. The adjuvant properties of mTNF alpha and pDNAmTNF need further investigation. PMID- 15125241 TI - [Sequence, necessary for initiating RNA synthesis, in the 3'-noncoding region of the classical swine fever virus genome]. AB - Classical swine fever virus (CSFV) is the causative agent of swine fever, which represents an economically important disease in hogs. We previously made a prediction about the recognition sites of replication initiation of CSFV by using the information content method, and it was predicted that the 21 nucleotides located at 3' end of the CSFV genome 3'UTR were essential to CSFV replication. In this paper, we experimentally studied these 21 nucleotides by site-directed mutagenesis. It was found that the 3'UTRs with the 21 nucleotides had the function of initiating RNA synthesis, while the 3'UTRs without the 21 nucleotides did not. The 21 nucleotides alone, without the rest of 3'UTR, were able to initiate RNA synthesis, though with a slump. It was demonstrated that the 21 nucleotides were essential to the replication of CSFV genome. The other part of 3'UTR was also required for sufficient RNA synthesis. It is highly likely that the 21 nucleotides were the necessary site for the CSFV genome replication initiation, and that the elements required for sufficient RNA synthesis were in the other part of 3'UTR. It was assumed that the CSFV replicase bound to the site and initiated the replication of the CSFV genome. In the 21 nucleotides, it was found that the mutation of position 216 and destruction of the 3' terminus in the 3'UTR precluded initiation of RNA synthesis, that the mutation of position 212 did not affect the capacity for initiation of RNA synthesis but attenuated the synthesis of RNA. Among the four mutants of 3'UTR at position 219, three produced the 3'UTR without initiation of RNA synthesis, and the other one produced the 3'UTR with initiation of less RNA synthesis. Therefore, it could be concluded that T216 was the most important while T212 was the least important, and that G219 and C228 were also important for RNA synthesis. The normal base component within the 21 nucleotides was essential to sufficient RNA synthesis. PMID- 15125243 TI - [Key events in the cell cycle, their regulation and organization]. AB - The review surveys the studies of molecular genetic mechanisms of the cell cycle control on various eukaryotic models. The major cell cycle phenomena are considered: (1) checkpoints and their role in preserving DNA integrity and fidelity of mitosis, (2) the cell oscillator model, and (3) the role of cyclins in timing of cell division and coordination of mitotic events. The main classes of regulatory proteins involved in the cell cycle are discussed in detail. PMID- 15125244 TI - [Use of genetically modified embryonic stem cells for creating transgenic animals]. AB - Targeted genetic modification of embryonic stem cells (ESC) was used to obtain nondifferentiated cell clones containing the foreign genetic material in the genome. It was demonstrated that transgenic animals may be obtained by ESC injection in preimplantation embryos and subsequent transplantation of the embryos into a recipient female. Using this method, we constructed chimeric animals with a modified genome. PMID- 15125245 TI - [Cause for maintaining high instability at the yellow gene in Drosophila melanogaster lines, isolated during the "mode for mutation" period in Uman populations]. AB - Mobile genetic elements are responsible for most spontaneous mutations in Drosophila melenogaster. The discovered in the 1980s phenomenon of frequent change of the wild-type yellow phenotype for a mutant one, and vice-versa, in strains of Drosophila melanogaster isolated from the Uman' natural population can be, according to our data, explained by repeated inversions and reinversions of the gene regulatory region located between the two copies of the hobo transport. However, most molecular genetic events accompanying the process can occur without the phenotype change. After several generations, the strains, remaining phenotypically unchanged, can possess different molecular genetic properties with respect to yellow. Using genetically homogenous or isogenic strains for the genetic analysis or for production of the new plant cultivars or animal breeds, geneticists and breeders often face the problem of stability of the strains. In the present study, the mechanism underlying the generation of instability at the yellow locus of D. melanogaster determined by the hobo-induced genome instability is described. PMID- 15125246 TI - [Change in nucleotide sequence of mitochondrial DNA from cytochrome b gene in chars of Salvelinus genus]. AB - Nucleotide sequences of two (405- and 1050-bp) regions of mitochondrial DNA (mtDNA) cytochrome c gene were established in chars of the genus Salvelinus from Russian Far East and Siberia. Based on the divergence and phylogenetic analysis of nucleotide sequences of the mtDNA cytochrome c gene, S. laecomaenis was shown to carry the most ancient mitochondrial lineage, which is close to the ancestral one. The archaic mtDNA of S. levanidovi occupied an isolated position on the phylogenetic trees. The mtDNA lineage of the southern S. malma was close to the S. alpinus-S. malma complex group. Within the S. alpinus-S. malma complex, three groups of mtDNA types having particular geographic distributions were distinguished. The Kolyma-Chukotka group includes lake S. taranetzi, S. boganidae and S. elgyticus from Chukotka, lake chars from Kolyma. The Okhotsk group is represented by northern S. malma, lake chars from northern Sea of Okhotsk, and anadromous S. taranetzi. The Siberian group is close to the Okhotsk one and consists of Taimyr and Baikal region chars as well as Arctic char from Finland. The divergence of char mitochondrial lineages was dated to the Pliocene Pleistocene. PMID- 15125247 TI - [Structure and expression of the F6.2 gene in Chironomus thummi and other Chironomus species]. AB - A full-length copy of the F6.2 gene from the tissue-specific BRa locus of the Chironomus thummi chromosome IV was isolated and analyzed. The gene contains two exons (715 and 644 bp, respectively) and one 172-bp intron. The data of the RT PCR analysis demonstrated that F6.2 was transcriptionally active at different developmental stages of Chironomus thummi and at least in the last larval stage of C. dorsalis. The distribution of the F6.2 gene among 42 species of Chironomus, as well as among two other genera of the family Chironomidae was examined by means of PCR. The F6.2 sequence was found in 34 Chironomus species. Using in situ hybridization, three species were analyzed for the presence of the F6.2 homologous sequences. In five species, the sequence of the F6.2 PCR product was determined. In these species, the intron size polymorphism caused by the variation of the number of the intron-forming repeats was observed. The data obtained provided evaluation of the F6.2 distribution among the genus Chironomus. PMID- 15125248 TI - [Multiyear monitoring of the ecological status of industrial zones of Moscow using a test line of Drosophila melanogaster]. AB - For 12 years, recombination rate was assessed in Drosophila melanogaster males exposed on a thermoelectric power station (TPS) of Moscow. In 1994, experiments were also carried out on another Moscow TPS for comparison. The recombination frequency in exposed males was two- or threefold higher than in the control sample. Recombination frequencies observed in different years did not significantly differ from each other. Likewise, no significant difference was observed for the results obtained on two TPSs. A dramatic increase in recombination frequency in flies exposed on TPS was considered as an adequate response to high concentrations of effective mutagens discharged by TPS. PMID- 15125249 TI - [Mutation of Drosophila melanogaster glass-like suppress expression of mini-white transgenes and dependence on their genomic environment]. AB - Pleiotropic recessive mutation glass-like (gl-l) found in region 8C-10E of the X chromosome was shown to cause glass-like eyes having no boundaries between facets and a nonuniform pigment distribution in the presence of the endogenous white gene. The gl-l mutation completely inhibited expression of the mini-white transgene contained in several constructs, but the effect depended on the site of construct integration in the genome. The mutation had no effect on the expression of the white transgene having the enhancer and flanked by insulators. The gl-l mutation did not affect the extent of mosaic eye pigmentation when a construct with mini-white was inserted in the telomeric or pericentric region. However, in most cases it completely inhibited the mosaic mini-white expression when cloned heterochromatic repeats were adjacent to the reporter gene in a construct. The gl l gene was assumed to play a role in the formation of the chromatin structure, because the effect of its mutation on expression of the white transgene depended on the transgene insertion site, the presence of insulators or an enhancer in the vicinity of the transgene, and on the adjacent heterochromatic repeats. PMID- 15125250 TI - [Genes controlling ontogenesis: morphosis, phenocopies, dimorphs, and other visible expressions of mutant genes]. AB - We studied facultative dominant lethal mutations obtained earlier in Drosophila melanogaster. In some genotypes, these mutations were expressed as lethals, but in other genotypes they lacked this expression. The mutations were maintained in the following cultures: (1) females Muller-5 heterozygous for the mutation; (2) males crossed to attached-X females; and females and males homozygous for the mutation. During culturing, many mutations were found to give rise to phenotypically abnormal progeny. Generally, these abnormalities were morphoses involving various body parts; they were mostly asymmetric and non-heritable. Maternal and paternal effects in the formation of morphoses were observed. In four cases, dimorphic mutations were recorded: a female homozygous for the mutation had mutant phenotype whereas its male counterpart was phenotypically normal. The mutations were recessive with regard to the norm. New phenotypes behaving as mutations with incomplete penetrance arose during culturing. In cultures of mutant homozygotes phenocopies would appear en masse; they would persist for one or two generations and disappear. One wave of phenocopies succeeded another. Visible phenotypes appeared, which further behaved as ordinary recessive mutations. We concluded that these visible manifestations are characteristic for regulatory mutations controlling ontogeny. Their appearance is explained by the activation of new regulatory scenarios caused by blocking standard regulatory pathways. PMID- 15125251 TI - [Evaluation of the combined ability of triticale and secalotriticum by signs of androgenesis in vitro]. AB - The combining ability of cultivars and strains of triticale and secalotriticum with respect to the characteristics of androgenesis in anther culture has been analyzed (the 6 x 3 polytester design and parental forms). Nonadditive effects of gene interaction have been found to be prevalent in the genetic control of the characters analyzed in plants with the given genotypes. However, the characteristics of androgenesis were the highest in those hybrid combinations in which genotypes with high values of general combining ability (GCA) and positive effects of specific combining ability (SCA) participated. A program of the analysis of combining ability in triticale and secalotriticum is proposed. The program is aimed at detecting hybrid combinations with the best androgenetic capacities and takes into account both additive and nonadditive effects. PMID- 15125252 TI - [Analysis of DNA markers specific for G-genome of wheat]. AB - A RAPD marker specific for the G genome of wheat was identified. The corresponding 1171-bp DNA sequence was cloned and analyzed. Screening of the database did not reveal any homologies with the known plant DNA sequences. Using the primers specific to the flanking regions of the marker sequence, PCR analysis of the polyploid wheat species and the diploid species of the section Sitopsis was carried out. In addition, using the cloned sequence as a molecular hybridization probe, RFLP analysis of the genomic DNA of these species was performed. PMID- 15125253 TI - [Genetic characteristics and prognosis of the existence of the European bison free-living population created in the "Orlovsky Poles'e" National Park]. AB - We present preliminary data on genetic aspects of preserving viability of the free-living population of European bison (Bison Bonasus L.) created in the Orlovskoe Poles'e National Park. PMID- 15125254 TI - [Differentiation of Salvelinus malma charr and Taranetz Charr Salvelinus taranetzi from PCR-RFLP analysis of mitochondrial DNA]. AB - Genetic differentiation of two sympatric charr species, the Dolly Varden charr (Salvelinus malma Walbaum) and the Taranetz charr (S. taranetzi Kaganovski), has been studied. Restriction fragment length polymorphism (RFLP) analysis has been used to compare three mitochondrial DNA (mtDNA) fragments (ND1/ND2, ND5/ND6, and Cytb/D-loop) amplified via polymerase chain reaction (PCR). The divergence between S. malma and S. taranetzi inferred from mtDNA nucleotide sequences is 2.8%; between S. leucomaenis and S. taranetzi, 7.1%; and between S. malma and S. leucomaenis, 7.5%. The absence of common haplotypes and the degree of divergence indicate that the Dolly Varden charr and the Taranetz charr are genetically isolated and confirm that S. taranetzi may be regarded as a separate species. PMID- 15125255 TI - [Acclimatization of salmon Oncorhynchus gorbuscha (Walbaum) in the European north: data from restriction analysis of mtDNA]. AB - Pink salmon spawners introduced into the White Sea basin (the Umba River) were compared to the spawners from the basin of the Sea of Okhotsk (the Ola River) using restriction analysis of two fragments of mitochondrial DNA (mtDNA). One of the fragments included genes ND5/ND6, the other, the cytochrome b gene and the control region. It was found that mtDNA variation and diversity at the earlier examined nuclear allozyme genes significantly decreased in the odd broodline of pink salmon 8 years after the introduction. The haplotype diversity in the even broodline was considerably lower than in the odd broodline exhibiting virtually no change two generations after the introduction. Based on the results obtained, a possible role of these changes in adaptation of White Sea pink salmon from the odd broodline to the new environment is discussed. PMID- 15125256 TI - [Polymorphism of glutathione-S-transferase M1 and P1 genes in patients with cystic fibrosis and chronic respiratory tract diseases]. AB - Polymorphism of GSTM1 and GSTP1 genes was studied in patients with cystic fibrosis (CF) and chronic bronchopulmonary diseases (CBPD) living in Bashkortostan. A combination of certain GSTM1 and GSTP1 genotypes accompanied by severe mutations in CFTR gene proved to intensify a pathologic process in respiratory organs of patients with CF; a combination of the normal GSTM1 and heterozygous I/V GSTP1 genotypes is the most favorable (OR = 4.49; chi 2 = 11.53, P < 0.002). In patients with CBPD, a combination of the GSTM1 null genotype and the homozygous GSTP1 V/V genotype is the most common (5.5% versus 1.3% in control; chi 2 = 3.01, P = 0.08). The frequency of this genotype is highest in groups of patients with recurrent bronchitis (8.1%; P = 0.07; OR = 6.75) and bronchiectatic disease (BED) (9.1%, P > 0.10, OR = 7.65). A combination of the null GSTM1 and I/V GSTP1 genotypes was found in 40.0% of patients with chronic nonobstructive bronchitis (chi 2 = 4.87; P = 0.03; OR = 4.03). Among patients with BED, a proportion of individuals with the normal GSTM1 and I/V GSTP1 genotypes was increased (36.4% versus 19.4% in control). In patients with chronic obstructive pulmonary disease (COPD), the frequencies of the GSTM1 and GSTP1 genotype combinations virtually did not differ from those in the control group suggesting that COPD severity is not related to changes in activities of glutathione S-transferases M1 and P1. PMID- 15125257 TI - [VNTR-polymorphism of the PAH, e-NOS genes and deletion of the CCR5 gene in populations in the northern Caucasus]. AB - VNTR allelic polymorphism at the phenylalanine hydroxilase (PAH) and endothelial constitutive nitric oxide synthase (eNOS) genes and the prevalence of the CCR5 chemokine receptor gene 32-bp deletion were examined in four indigenous populations of Northern Caucasus, Adygs, Kumyks, Karachais, and Nogais (Kuban and Karanogais). Population-specific features of the allele and genotype frequency distribution patterns of the polymorphisms examined were described. The data obtained were compared to those obtained from literature. The results of the study confirmed that the frequency and occurrence of the PAH polymorphic alleles exhibit substantial interpopulation differences. In the populations of Northern Caucasus, the eNOS minisatellite polymorphism alleles and genotypes frequency distribution patterns were close to those described earlier for populations of the Volga-Ural region (VUR), and also for the Australian Caucasoids, Japanese, and Turks. In the populations examined, the mean frequency of the CCR5 gene deletion was 0.055, which was somewhat lower than in the populations of VUR (0.07) and Europe (0.081), and practically identical to that in Asian populations (0.050). For each population observed and expected heterozygosities at each locus were calculated. In general, the gene pool of Northern Caucasian populations showed substantial differentiation at the loci examined: the GST value was 0.0274. The data for individual loci showed that the greater contribution to the interpopulation diversity was made by the differences in the PAH VNTR allele frequencies (GST = 0.04), while the differences at the eNOS and CCR5 loci were small (GST = 0.0025 and GST = 0.0039, respectively). PMID- 15125258 TI - [Structure of the gene pool of eastern Ukrainians from Y-chromosome haplogroups]. AB - Y chromosomes from representative sample of Eastern Ukrainians (94 individuals) were analyzed for composition and frequencies of haplogroups, defined by 11 biallelic loci located in non-recombining part of the chromosome (SRY1532, YAP, 92R7, DYF155S2, 12f2, Tat, M9, M17, M25, M89, and M56). In the Ukrainian gene, pool six haplogroups were revealed: E, F (including G and I), J, N3, P, and R1a1. These haplogroups were earlier detected in a study of Y-chromosome diversity on the territory of Europe as a whole. The major haplogroup in the Ukrainian gene pool, haplogroup R1a1 (earlier designated HG3), accounted for about 44% of all Y chromosomes in the sample examined. This haplogroup is thought to mark the migration patterns of the early Indo-Europeans and is associated with the distribution of the Kurgan archaeological culture. The second major haplogroup is haplogroup F (21.3%), which is a combination of the lineages differing by the time of appearance. Haplogroup P found with the frequency of 9.6%, represents the genetic contribution of the population originating from the ancient autochthonous population of Europe. Haplogroups J and E (11.7 and 4.2%, respectively) mark the migration patterns of the Middle-Eastern agriculturists during the Neolithic. The presence of the N3 lineage (9.6%) is likely explained by a contribution of the assimilated Finno-Ugric tribes. The data on the composition and frequencies of Y chromosome haplogroups in the sample studied substantially supplement the existing picture of the male lineage distribution in the Eastern Slav population. PMID- 15125259 TI - [Search for transcribed segments in the region of q14.3 of human chromosome 13 in silico]. AB - Using computer-aided genomic methods, a complete map of the expressed sequence tags (EST) located in the human genome region 13q14.3 between the STS markers, D13S810 and D13S1469, was constructed. A total of 62 EST clusters were formed, of which 12 clusters corresponded to the already known human genes, 4 clusters represented pseudogenes, and 10 clusters were new human genes. The use of the method of reverse transcription in combination with polymerase chain reaction (RT PCR) provided experimental confirmation of the existence of mRNA transcripts for the novel human genes revealed in silico. PMID- 15125260 TI - [Zoia Sofron'evna Nikoro. On the 100th anniversary of her birthday (9.02.1904 13.10.1984)]. PMID- 15125261 TI - [60th. anniversary of the Mexican Heart Institute]. PMID- 15125262 TI - [Arrhythmic activity of the papillary muscle induced by high frequency stimulation: n1 rhythms,transition forms and hysteresis]. AB - Cardiac tissues are able to work within a wide range of frequencies to respond to the changing requirements an organism may have. However, during these frequency variations and under certain pathologic conditions arrhythmias such as blocks, tachycardia, fibrillation, etc, may arise some with fatal consequences. For this reason several experimental procedures have been developed that have shown to be useful in studying whole heart properties, or as an alternative from portions of it when changes in its work rate are imposed. This study reports different phenomena occurring in the papillary muscle of the guinea pig heart when stimulated at very high frequency, of several tens of pps, while analyzing its responses during gradual increments starting at 1 (pulses per second). We found that in our conditions papillary muscles display N:1 rhythms with progressive higher N; further more we found that between one and the next rhythm diverse transition patterns appear, among them a new one that we have named "burst pattern". Finally we show that our system exhibits a generalized process of hysteresis by frequency, being this the first report for guinea pig cardiac tissue and the first one to show also the presence of several hysteresis loops in the same experiment. Due to the large volume of generated data we used a faster and easier way to analyze and display them, based on the fast Fourier transform (FFT). The method is briefly described. PMID- 15125263 TI - [Correlation between left ventricular ejection fraction assessed by gated SPECT and radionuclide ventriculography in patients with ischemic cardiomyopathy]. AB - BACKGROUND: Recently, the use of Gated SPECT to assess ejection fraction, volumes and ventricular wall motion has been suggested. The aim of this study is to compare the ejection fraction (EF) obtained by Gated SPECT (QGS, Cedars Sinai) with the values obtained through equilibrium ventriculography in patients with ischemic heart disease. METHODS: We studied 50 consecutive patients that were diagnosed by equilibrium radioisotopic ventriculography and Gated SPECT with a dual protocol (TI-201 rest/Tc-99m SestaMIBI stress) using the QGS, Cedars Sinai software. RESULTS: We found a 92% correlation between both tests. In the subgroup analysis, the higher correlation (r = 0.89) was found in group 2, which included patients with moderate ventricular dysfunction (EF 30-50%), whereas, in patients with severe ventricular dysfunction (group 3, EF < 30%), the correlation was lower (r = 0.76) than in group 2, but higher than in patients with EF > 50% (group 1). CONCLUSIONS: There is a good correlation between EF obtained with Gated SPECT and equilibrium ventriculography, regardless of the EF value. PMID- 15125264 TI - [Assessment of diastolic function in Chagas' disease with pulsed doppler tissue imaging]. AB - Diastolic function is involved early during the undetermined form of Chagas' disease (Ch). Pulsed Doppler tissue imaging (DTI) is a new technique to evaluate diastolic function recording myocardial velocities in the longitudinal axis. To evaluate the relevance of DTI in patients with Ch, we studied, with Doppler and echocardiography, 49 patients (average age 49 +/- 12 years) and 20 normal subjects (average age 45 +/- 15 years) as a control group (C). Patients were divided in four groups according to the pattern of their mitral and pulmonary vein flows: Normal diastolic function (N), prolonged relaxation (PR), pseudonormal (PN) and restrictive (R). Peak velocity of the E wave of the lateral mitral annulus (Ea) was diminished in PR, PN, and R patients (0.11 +/- 0.02 m/s, 0.10 +/- 0.02 m/s and 0.12 +/- 0.06 m/s, respectively) as compared to group C (0.18 +/- 0.07 m/s, p < 0.01) but not with respect to group N (0.15 +/- 0.03 m/s, N.S.). In this latter group only peak E velocity of transmitral flow/Ea ratio was useful to detect diastolic abnormalities as compared to group C (4.92 +/- 0.98 vs 4.14 +/- 1.26, p < 0.05). DTI was useful in the assessment of diastolic function in Ch, including those patients in whom mitral and pulmonary vein flows were normal. PMID- 15125265 TI - [Surgical treatment results of congenital heart defects in children with Down's syndrome]. AB - We made a retrospective study of patients with Down's syndrome that were surgically treated for correction or palliation for their congenital heart disease between january 1996 to december of the 2000 in the National Institute of Cardiology "Ignacio Chavez". We analyzed these variables: age at the surgical moment, sex, congenital heart defect, pulmonary arterial pressure, type of surgery, time of stay in the intensive care unit, complications and mortality. In this period they were surgically treated 37 patients. The mean age was of 2 years with 8 months with a range of 2 months to 17 years. The interventricular defect was the most frequent one (35%) and it was associated to persistent ductus arteriosus in the 61% of the cases. Six patients (16%) had atrioventricular septal defect, the half of them type A and other half type C of Rastelli classification. Twelve patients (32%) had one lesion persistent ductus arteriosus. Three patients had tetralogy of Fallot and two atrial septal defect. Pulmonary hypertension was found in 90% of the patients, in 23 was severe (62%), moderate in 5 and slight in 6 (16%). The surgical treatment was corrective in 89% patients and the average time of stay in intensive care unit was of 2.5 days. The most frequent complication was rhythm and conduction disorders, in 8 patients (22%), three with complete AV block and the mortality was of 8%. In the patients with Down's syndrome is important a complete clinical evaluation with an eye toward establishing an opportune surgical treatment. PMID- 15125266 TI - Multiple congenital coronary artery fistulae draining into the left ventricle. AB - Coronary artery fistula between a coronary artery and a cardiac chamber is a rare condition, especially when multiple fistulas communicate with the left ventricle. Herein we report a case of an elderly woman with multiple diffuse coronary artery left ventricular fistulas diagnosed by angiography. Since the coronary artery cardiac chamber communications were multiple and diffuse neither surgery nor transcatheter coil occlusion was considered in this case. PMID- 15125267 TI - [Giant intracardiac Rhabdomyoma in neonatal stage. Report of one case]. AB - Primary heart tumors are not very common, frequent ones during childhood are the rhabdomyomas. This diagnostic can be made since fetal life. Evolution variates, in some cases there are no symptoms and the diagnostic represents an echocardiographic finding, other cases are detected associated with tuberous sclerosis and a small group express by arrhythmias or low output. Because of the strange of this presentation this case is presented. PMID- 15125268 TI - [High density lipoproteins (HDL). A therapeutic objective in the atherosclerosis prevention?]. AB - High density lipoproteins (HDL) are a family of heterogeneous particles that vary in size, density and chemical composition, as a result of their synthesis and catabolism rates, and a continuous intravascular remodeling by the action of enzymes and transport proteins. Low plasma levels of HDL correlate with a high risk of atherosclerotic heart disease. Such a diminished concentration of HDL affect reverse cholesterol transport, which is the metabolic pathway responsible for the movement of cholesterol excess from peripheral tissues to the liver for recycling or excretion. In addition, HDL possess anti-inflammatory, anti oxidative, anti-aggregatory, anti-coagulant, and pro-fibrinolytic properties, as has been demonstrated by in vitro studies. Some of those potentially anti atherosclerotic in vitro-properties has been corroborated by HDL infusion in vivo. Such evidences and the protection of susceptible animals from atherosclerosis by transgenic manipulation of HDL metabolism, raise the possibility to focus the HDL plasma levels as a main target in coronary heart disease prevention. Intervention trials have shown an important reduction in coronary events by rising HDL-cholesterol, mainly in the secondary prevention. Increasing HDL plasma levels by hygienic intervention such as aerobic exercise, weight loss and stop smoking is strongly recommended to reduce coronary risk in primary prevention. Pharmacological intervention to rise the HDL plasma levels with niacin or fibrates, should be considered in some patients as an alternative when hygienic intervention fails. Finally, it most be taken into account that the different HDL subclasses does not possess the same anti-atherosclerotic properties, suggesting that hygienic and pharmacological interventions should focus to increase HDL functionality rather than HDL-cholesterol plasma levels. PMID- 15125269 TI - [Changes in cardiac metabolism and their possible therapeutic use. (part II)]. AB - Based on the cardiac metabolic changes during hypoxia, in this second part of our review we propose, the polarizing solution as an alternative for the maintenance of the cardiac cells during an infarction, in conjunction with other alternative therapies. PMID- 15125270 TI - [Mitral valve stenosis and regurgitation associated with atrial septal defect and organic tricuspid damage]. PMID- 15125271 TI - Carboplatin ototoxicity: an animal model. AB - A new animal model of ototoxicity is presented using intravenous carboplatin in adult chinchillas. A range of physiological and morphological effects was produced using doses calculated from the recommended therapeutic range (200-400 mg/m2). Auditory thresholds to tone pips stimuli were monitored using brain stem evoked responses (ABR). Cochlear histopathology was studied by light microscopy (LM) and ultrastructural hair cell abnormalities investigated with scanning electronmicroscopy (SEM). Carboplatin in this animal model predominantly affected the inner hair cells. This may provide an important model for the study of selective loss of the main afferent input in the auditory system. PMID- 15125272 TI - Audit of the incidence of persistent perforation of the tympanic membrane following T-tube removal or extrusion. AB - Audit of the incidence of persistent perforation of the tympanic membrane following T-tube removal or extrusion was carried out on 100 patients (151 ears). The incidence was found to be 11.9 per cent. This was independent of whether the tube was surgically removed or extruded; of the grade of surgeon carrying out the operation; and whether the patient treatment was carried out privately or on the N.H.S. Changes of practice have resulted from the audit. PMID- 15125273 TI - Audit of the incidence of persistent perforation of the tympanic membrane following grommet removal or extrusion. AB - The incidence of persistent perforation following grommet removal or extrusion in 210 ears was assessed, and found to be zero per cent. Collar button grommets were retained longer than Shepard grommets, and the percentage of ears requiring two or more tube insertions was 51.9 per cent. PMID- 15125274 TI - Hypersensitivity to topical corticosteroids in otitis externa. AB - Over a 12-month period, nine patients were seen in a contact dermatitis clinic with an inflammatory dermatosis localized predominantly to the external auditory canal. Of these patients, four were found to have an allergic contact dermatitis which was exacerbating their dermatitis. All were allergic to one or more corticosteroids and topical antibiotics used in the treatment of their dermatitis. We would suggest that hypersensitivity to topical corticosteroids and other medicaments be considered in patients with otitis externa which fails to respond to treatment. PMID- 15125275 TI - Analysis of lectin receptors in normal nasal mucosa, nasal polyp, inverted papilloma and papillary adenocarcinoma. AB - In order to investigate the changes in glycoprotein structure in the process of cellular differentiation of the nasal mucosa, formalin-fixed, paraffin-embedded biopsy specimens of normal nasal mucosae, nasal polyps, inverted papillomas and papillary adenocarcinomas were analysed by the Avidin Biotin-Peroxidase Complex technique for the demonstration of peanut agglutinin (PNA) receptors, concanavalin ensifomis agglutinin (ConA) receptors, ulex europeaus agglutinin (UEA-I) receptors, wheat germ agglutinin (WGA) receptors, carcino-embryonic antigen (CEA) and keratin. The quantity and distribution of PNA receptors, ConA receptors, UEA-I receptors and CEA were different, in relation to the varying pathological changes. The results suggest that the glycoprotein structure in the cells of the nasal mucosa will change following their differentiation and malignant transformation, which may be helpful in establishing the diagnosis. PMID- 15125276 TI - Alteration of airflow and mucociliary transport in normal subjects. AB - The effect that variation of nasal airflow has on the mucociliary clearance was studied in healthy volunteers. The nasal saccharin particle clearance time was found to have an inverse relationship with the nasal airflow. PMID- 15125277 TI - Steroids and control of post-tonsillectomy pain. AB - A prospective, randomized, double-blind study to assess the effect of steroids on post-tonsillectomy pain was performed on 82 adults. The premedication, anaesthesia, surgical technique and post-operative analgesia were standardized. Pain was assessed on a visual analog scale. Steroids were found to have no appreciable effect on the amount of post-operative pain. PMID- 15125278 TI - CO-N reaction--a new serological activity index--on Wegener's granulomatosis. AB - Cordyceps ophioglossoides is one of the Japanese old 'Kanpoh' drugs used for metrorrhagia as a decoction. We found that CO-N, a galactosaminoglycan from Cordyceps ophioglossoides, reacted with sera from patients with some collagen diseases. By using CO-N, we made a new serological activity index (CO-N reaction). In the present study, we investigated CO-N reaction on patients with Wegener's granulomatosis (WG). The aggregation titres of CO-N (CO-N numbers) displayed a possible correlation with their clinical activity. CO-N reaction might also serve an important role in supporting the diagnosis of active WG and in helping to assess the degree of disease activity. The purpose of this report is to introduce this new serological activity index for Wegener's granulomatosis. PMID- 15125279 TI - The treatment of primary recurrence following laryngectomy for laryngeal carcinoma. AB - Of the 397 patients undergoing total laryngectomy for squamous cell carcinoma by Professor P. M. Stell between 1963 and 1991, 73 are known to have suffered a local recurrence. Of these, 17 were treated by radiotherapy and/or further surgery. Secondary surgery was reserved for selected cases of peristomal and pharyngeal recurrence: of the 35 peristomal recurrences, eight were treated surgically. Previous reports of recurrence after total laryngectomy have focused on the problem of peristomal recurrence. Patients with pharyngeal recurrences can also be treated satisfactorily with microvascular surgical reconstruction techniques. Though rarely curative, secondary total pharyngectomy can be highly palliative and results in little additional morbidity. PMID- 15125280 TI - Cervical lymphadenopathy due to mycobacterial infection: a diagnostic protocol. AB - Mycobacterial cervical lymphadenopathy is relatively uncommon in the United Kingdom; when cases do occur opportunities for early diagnosis and treatment may be missed. We have reviewed twenty-three cases of mycobacterial cervical lymphadenopathy presenting to an urban general hospital over a four-year period. We discuss the techniques available to aid a diagnosis of mycobacterial disease and suggest a protocol to allow efficient use of these techniques. PMID- 15125281 TI - Eye protection in ear, nose and throat surgery. AB - The aim of this study was to quantify the risk of blood splashes to the conjunctiva during ear, nose and throat surgery. Two hundred and sixty consecutive operations by three surgeons were assessed for contamination of safety glasses and other factors. We found that 15 per cent of operations resulted in blood droplet contamination of safety glasses. In 92 per cent of these, contaminations were on the exterior of the glasses and eight per cent were on both sides. We concluded that about one and a half per cent of operations would result in droplet contamination of the conjunctiva and that safety spectacles reduced the risk by a factor of about 10. This study generally concurs with that of previous research in the areas of general and orthopaedic surgery and necropsies. We substantiate the need for eye protection for all ear, nose and throat procedures. Spectacles do provide a reasonable degree of protection but where absolute protection is needed i.e. in high risk groups, goggles should be used in preference to safety spectacles. PMID- 15125282 TI - Hearing loss fluctuating with blood sugar levels in Meniere's disease. AB - An unusual case of a patient with bilateral Meniere's disease is described whose disease presented in the second ear as a sensorineural hearing loss which fluctuated with the patient's level of blood glucose. The literature concerning the role of abnormal glucose metabolism in Meniere's disease is reviewed and the investigation and management of this patient's condition is discussed. PMID- 15125283 TI - Acquired aphasia without deafness in childhood--the Landau-Kleffner syndrome. AB - A young boy presented with loss of speech and behaviour disturbance and was thought to be deaf. He was subsequently found to have the Landau-Kleffner syndrome (LKS), or acquired aphasia with epilepsy. Children with this disorder commonly present to an audiology or ENT clinic. Early recognition is important to initiate supportive, speech and educational care. PMID- 15125284 TI - A variation of first branchial cleft anomalies. AB - First branchial cleft anomalies are considered to be duplications of the external auditory meatus (EAM) and pinna with a sinus that runs parallel to the EAM (Type 1) or with a sinus that runs from an opening in the neck and ends blindly near the cartilaginous EAM (Type 2). In this paper we discuss a young patient that presented with an infected sinus that did not resemble either of the two known types of first branchial cleft anomalies. PMID- 15125285 TI - Acute osteomyelitis of the maxilla in the newborn. AB - Acute osteomyelitis of the maxilla in the new born is a rare infective condition of the maxilla which subsequently spreads to include the eye, nasal and oral cavities with their attending signs and symptoms. Possible sequelae include death, ophthalmological, laryngological and dental complications. The organism responsible is usually Staphylococcus aureus and early diagnosis and treatment can result in rapid resolution of the condition. PMID- 15125286 TI - Prostatic metastases in the nose and paranasal sinuses. AB - Prostatic metastases in the nose and paranasal sinuses are rare. Seven cases have previously been reported in the world literature. We describe the clinical presentation of a patient with prostatic metastases and the use of prostate specific antigen in confirming the diagnosis. We also review the literature about metastases involving the nose and paranasal sinuses. PMID- 15125287 TI - Bony remodelling in an osteoma of the paranasal sinuses. AB - Osteoma is the commonest benign tumour of the paranasal sinuses (Handousa, 1952). A case of a large osteoma removed from the frontal and ethmoid sinuses is presented. Histopathological examination of this tumour revealed evidence of bony remodelling similar to that seen in Paget's disease which has not been previously reported. The pathological significance of this finding is discussed. PMID- 15125288 TI - Recurrent laryngeal papillomatosis: a case of florid papillomatosis following a remission of 30 years. AB - Laryngeal papillomatosis is the commonest benign tumour affecting the larynx. Two forms are found i.e. juvenile onset and adult onset. Typically the juvenile onset form has a greater rate of recurrence and often remits with the onset of puberty (Corbitt et al., 1988). The human papilloma virus (HPV) is the causative agent (Abramson et al., 1987; Corbitt et al., 1988), specifically types HPV6 and 11. Attempts have been made to correlate the clinical behaviour of these two modes with the viral serotype and other aetiological factors such as smoking and hormonal factors (Abramson et al., 1987; Rimmel et al., 1992). Studies, however have shown that there is considerable variation in behaviour (Steinberg et al., 1983; Corbitt et al., 1988; Crissman et al., 1988). It is widely accepted that the disease 'burns' itself out, particularly with respect to the juvenile form. It is interesting and unusual therefore when the disease reappears after many years of remission. The following case report illustrates this point. PMID- 15125289 TI - Cough and stridor: who should investigate the patient? AB - Stridor is usually produced by obstruction in the upper airways. We present a case of stridor referred to the ENT Department in whom an endoscopic examination as far as the lower trachea showed no abnormality. A subsequent bronchoscopy in the Chest Department revealed a tumour in the right main bronchus. PMID- 15125290 TI - Parapharyngeal space tumour presenting as recurrent uvular oedema. AB - Neoplasms of the parapharyngeal space are uncommon and usually present as an intra-oral or neck mass. They often elude early diagnosis due to their deep seated nature. Here we report a case presenting with recurrent oedema of the uvula. The pathophysiology of this previously unreported mode of presentation is discussed. PMID- 15125291 TI - Chondrosarcoma of the hyoid bone. AB - Chondrosarcoma of the head and neck is relatively rare. We report the sixth case known to us in which the chondrosarcoma had originated in the hyoid bone. The patient, a 66-year-old man, underwent surgery. The resected tumour was a 5 cm encapsulated lesion composed of lobulated, firm, gray-white tissue. Histologically it showed mature chondrocytes without capsular invasion. There was high cellularity and a few binucleated chondrocytes. Although the tumour grade was prognostic significance, our literature review of the five other cases suggests that the location of the primary lesion and the adequacy of its resection to be more important. Our patient continues to do well 15 months post operatively. However, long-term follow-up is essential as recurrences have been reported even after eight or 10 years. PMID- 15125292 TI - Emergency tracheostomy in a patient with Melnick-Needles syndrome and sleep apnoea. AB - A six-year-old girl with a rare bone dysplasia (Melnick-Needles Syndrome) presented with a five month history of severe sleep apnoea, weight loss and failure of thrive. The syndrome is associated with craniofacial abnormalities, including micrognathia. Following a multi-disciplinary assessment an elective tracheostomy was considered the most appropriate treatment. The patient developed severe respiratory distress 10 days prior to the arranged date of surgery and required an emergency tracheostomy. This resulted in a dramatic return to health. The recognition of severe sleep apnoea in patients with craniofacial abnormalities and the role of initial tracheostomy are discussed. PMID- 15125293 TI - Spontaneous retropharyngeal haematoma: two cases and a review of the literature. AB - Retropharyngeal haemorrhage is a rare condition. The classical picture is described as a triad of features; superior mediastinal obstruction, anterior displacement of the trachea on a plain X-ray of the neck and subcutaneous bruising appearing on the neck and spreading on to the chest wall (Sandor and Cooke, 1964). Two cases are reported here, neither of which had mediastinal compression. The available literature is summarized. PMID- 15125294 TI - Pathology of the eustachian tube in otitis media: an electron microscopic study. AB - The ultrastructure of the mucosa of the eustachian tube was studied in four temporal bones showing tympanosclerosis, cholesteatoma, otitic meningitis and a grafted tympanic membrane (tympanoplasty). The mucosa of tube was abnormal in the four cases confirming the relationship between the state of the eustachian tube and the inflammatory process in the middle ear. The observed abnormalities included: ciliary loss, abnormal ciliary morphology and motility, oedema of the microvilli, hyperplasia of the goblet cells and the seromucinous acini, desquamation of the non-ciliated cells and appearance of mast cells in the lamina propria of the tube. Ciliary changes were the most frequent abnormalities and the morphological changes, in general, were fewest in the case of healed tympanoplasty. The pathophysiology of the morphological changes was discussed and correlated with the disease in the middle ear. PMID- 15125295 TI - Post-surgical necrotizing palisading granuloma of the nose. AB - The finding of granulomata in nasal polyps is rare. In this report we describe a case with necrotizing granulomata occurring after surgery. The differential diagnosis of such a finding is discussed. PMID- 15125296 TI - Paragangliomas of the larynx. PMID- 15125297 TI - Paragangliomas of the larynx. PMID- 15125298 TI - The value of head dressings for middle ear surgery. PMID- 15125299 TI - Acute tonsillectomy in the management of infectious mononucleosis. PMID- 15125300 TI - Diagnostic laryngoscopy and bronchoscopy aided by the laryngeal mask airway. PMID- 15125301 TI - Disease management and Medicaid. PMID- 15125302 TI - Comprehensive reconstructive surgery for obstructive sleep apnea. AB - Obstructive sleep apnea (OSA) is commonly treated by continuous positive airway pressure (CPAP). Patients can be intolerant, noncompliant, or avoidant of CPAP therapy but incur increased risk of cardiopulmonary disease, stroke, and shortened life span without treatment. Comprehensive hard and soft tissue surgical therapy directed at the nasal, retro-palatal, and retro-lingual airway has been shown to optimize surgical outcomes compared with isolated soft tissue procedures. Outcome information from postoperative sleep studies shows that staged surgical procedures of hard and soft tissues are comparable to CPAP studies. While CPAP is the "gold standard" therapy for obstructive sleep apnea, combined soft tissue and maxillofacial surgical techniques can provide a treatment alternative for patients that are unable or unwilling to use CPAP treatment. PMID- 15125303 TI - Analysis for space-time clustering of CJD cases, Kentucky, 1988-1997. AB - Creutzfeldt-Jakob Disease (CJD) is a transmissible spongiform encephalopathy of humans. It can be transmitted iatrogenically or inherited as an autosomal dominant trait. However, most cases (85%) of the disease are sporadic, occurring stochastically in approximately one person per million population. In 1996, a new, neuropathologically distinct variant of CJD was recognized in a group of unusually young patients who had apparently been infected by exposure to the agent of bovine spongiform encephalopathy (BSE) via contaminated beef products. This prompted many countries, including the United States, to renew their surveillance for CJD to assess the possibility of other variant cases. Spatiotemporal aggregation of cases may indicate a common source of infection or some other shared risk factor. In order to assess the possibility of variant cases, death certificate data was used to examine the spatiotemporal patterns of CJD mortality in Kentucky over a ten-year period. Space-by-time clustering was assessed using Cluster 3.1 software. The findings of this study produced some evidence for a cluster of six cases in central Kentucky. Overall, however, there was little or no evidence to suggest the occurrence of variant or unconventionally acquired cases. PMID- 15125304 TI - Why BMI? PMID- 15125305 TI - A cancer clip. PMID- 15125306 TI - [Dear Reader of Neuropsychopharmacologica Hungarica]. PMID- 15125307 TI - [Antidepressive pharmacotherapy: changing paradigms]. PMID- 15125308 TI - [Cardiac effects of antipsychotics: mechanism of arrhythmias and sudden cardiac death]. AB - The review summarizes experimental and clinical data showing the cardiac side effects of antipsychotic drugs. Some antipsychotics may correlate with prolongation of QT interval, induce ventricular tachycardia, torsades de pointes, TdP, and sudden death. The author surveys the cellular actions of the drugs, the electrophysiological mechanisms and the recent data referring the drug's effects on ionic currents, mainly potassium currents. Most antipsychotics are associated with the inhibition of delayed rectifier K+ channels. Comparing the potency on K+ channel inhibition and the prolongation of the QT interval with the therapeutic plasma levels of the drugs, the difference between the inhibitory potency and the therapeutic dose is the highest in the case of quetiapine, olanzepine and risperidone, while thioridazine shows the smallest difference. All drugs that cause TdP prolong the QT interval and inhibit the K+ rectifier channel, but the relationship is not precise. Some additional cellular effects of particular agents, modulating conditions, factors (diseases, electrolytes disturbances, genetic damage, drug interactions) make the individual vulnerable to arrhythmia. The paper highlights drug interactions causing risk of arrhythmia during chronic treatment of psychiatric patients. PMID- 15125309 TI - [Pharmacotherapeutic trends at the beginning of the millennium in Hungary. Pharmacotherapy for bipolar patients, I]. AB - The therapy of bipolar disorders is a challenge for clinicians not only in the acute, but also in the maintenance phase. The choice is hazardous in spite of the many drugs available for this purpose. There are the classical mood stabilizers, antipsychotics, antidepressants, anxiolytics, hypnotics and other medicines that are usable in the treatment, but the resolution of the clinician can significantly influence the actual and prospective condition of the patient. We have evidence based experimental, anecdotal knowledge about the therapy of bipolar disorders, but we do not know the facts of the practice. The aim of the study was to evaluate the current therapeutic trends in Hungary. METHOD: A questionnaire was compiled and sent to psychiatrists working in different regions of Hungary. The questions referred to the institute, the patient (demographic data, actual condition, disorder) and the actual treatment. The study was blind to the investigator: nor the psychiatrists neither the patients were identified. The study covered patients visiting psychiatrists from 25 November to 6 December 2002. RESULTS: 204 questionnaires were evaluated. 82 male and 122 female patients were included, of the average age of 48 years. The distribution of the disorders was as follows: bipolar I: n = 120; bipolar II: n = 66; cyclothym: n = 18; and the actual condition was diagnosed, too. More than half of patients showed a balanced state. 28% of patients took only one, 35% two and 23% three kinds of psychotropic drugs, and 14% four or more simultaneously. 92% of patients was on "classical mood stabilizers" (lithium, carbamazepine, valproate) and the use of the carbamazepine was the most frequent. 54% took some kind of antipsychotic drugs and three quarter among them some atypical form. The use of anxiolytics was excessive: 44% of patients used one of them. 35% was on antidepressants and more than half of them on SSRI. CONCLUSION: The study findings show the current state of the treatment of bipolar disorders in Hungary, and are comparable with the relevant international trends and professional expectation. The results are useful in practice and in the education of physicians. PMID- 15125310 TI - [Experience with Risperidone in the treatment of institutionalized mentally retarded patients, with special reference to treatment of aggressive states]. AB - Aggression has become a problem of our everyday life; every psychiatrist meets aggressive patients in his practice almost daily. Aggression represents a special problem in the case of institutionalized, mentally retarded (severe and moderate) patients, when it is associated with agitation, deficit of critical functions, impulsiveness, mood disorders. The nursing staff of these institutions is often overworked; the affective outbursts and aggressive behavior of mentally retarded patients may provoke a hostile attitude on the part of the nursing staff towards the patients. In the case of mentally retarded patients, unpredictable events may occur at any time. The structural background of mental retardation, the function of the affected cerebral structures, is not completely clarified. It was found in several studies that risperidone is effective in the treatment of agitation and aggressive behavior; the incidence of side effects is much lower than in the case of typical antipsychotics. We started the treatment with risperidone of 60 mentally retarded patients; we evaluated the therapeutic outcome after a three month follow-up period using a rating scale made specially for this purpose. An attempt was made to compare the therapeutic results obtained in the risperidone group with the condition of patients receiving typical antipsychotics. It was found that, in the case of several items (aggression, agitation, deficit of critical functions, mood disorders, sleep disturbances, involvement in therapeutic activities), risperidone was significantly more effective than typical antipsychotics, and the incidence of extrapyramidal symptoms and other adverse events was much lower. It was hardly necessary to impose restraints in the risperidone group. In the care of mentally retarded patients, the use of risperidone has many long-term advantages, and hence it represents an effective alternative to typical antipsychotics. PMID- 15125311 TI - [Role of endogenous cannabinoids in cerebral reward mechanisms]. AB - Central cannabinoid receptors (CB1 receptors) are densely located in the output nuclei of the basal ganglia (globus pallidus, substantia nigra pars reticulata). Endogenous cannabinoids appear to modulate transmitter systems (e.g. dopamine) within the basal ganglia. In the striatum, CB1 receptors are localized on the same neurons as Gi-coupled dopamine D2 receptors. Striatal CB1 receptors are also negatively linked to adenylcyase, and may modulate dopamine release. The presence of CB1 receptors in dopaminergic neurons strongly suggests that cannabinoids play a modulatory role in dopaminergic neuronal pathways. This co-localization may postulate "cross talk" between endocannabinoids and dopamine-dependent reward mechanisms. PMID- 15125312 TI - [Janus-faced therapy: what is common in psychotherapy and pharmacotherapy?]. AB - The traditional biomedical approach separated psychotherapy and pharmacotherapy dichotomously. The biopsychosocial model of medicine stimulated the integration of these basic therapeutic modalities. There are many pieces of evidence testifying to the close relationship of somatic and psychological manifestations, and this connection is mainly represented by psychosomatic medicine. The present paper reviews some essential fields of psychotherapy and pharmacotherapy, e.g. similarities, interference, some important models of therapeutic integration, cost-benefit evidences, and some research implications. In today's medicine, integration is a basic requirement, and hence the dichotomous confrontation of psychotherapy and pharmacotherapy has become unacceptable. One important trend is the recognition that theories of modern psychotherapy cannot do without a sound knowledge of the brain functions. PMID- 15125313 TI - [Paralytic ileus during haloperidol therapy]. AB - In the case of elderly patients, the serious side effects of haloperidol should be taken into account. The 83-year-old schizophrenic patient had had successful haloperidol treatment, but developed a sudden paralytic ileus. The unique side effect of haloperidol calls attention to the danger of neuroleptic treatment of elderly patients. The good solution is to change over from haloperidol to a second-generation antipsychotic agent. PMID- 15125314 TI - About Meduna's pioneer activity. PMID- 15125315 TI - [Playing with fire? Depression and drug therapy]. PMID- 15125316 TI - [Significance of prognostic factors in oral squamous carcinoma]. AB - Despite the considerable advances in the diagnostic and therapeutic possibilities, the prognosis of epithelial tumors in the oral cavity is very poor. A knowledge of the prognostic factors at the beginning of treatment is therefore indispensable for determination of the appropriate therapy for the given patient. These factors may be linked to the patient (e.g. age, sex, general condition and immunological parameters) or to the tumor (localization, TNM stage, histological features, DNA content, or immunohistochemical and other parameters). A survey of the literature reveals that the TNM stage, the grade, the mode of invasion and the depth of the tumor are generally the most important factors influencing the fate of the patient. The prognosis primarily depends on the clinicopathological parameters, though even if they are known, it is not possible to screen out those patients who are at particular risk of a relapse. During the past 10 years, study of the DNA content, the proliferation markers and certain oncogenes has came into the focus of attention; great interest is also shown in the matrix metalloproteinases, which play key roles in the invasion and metastasis formation. PMID- 15125317 TI - [Revival of antihypertensive therapy: use of thiazide diuretics]. AB - In the international and the Hungarian guidelines, the diuretics in the first line of the treatment of hypertension. Their sometimes false judgment is based on the side effects, because of the over dosage of the applied medication. According to finished studies, efficiency of thiazides is usually the same as that of their competitors in influencing of the cardiovascular morbidity and mortality. Thiazides have to be given in the first line an antihypertensive treatment to the patients, especially if they are old, or have a great risk for a cardiovascular complication (stroke, coronary heart disease, heart failure, left ventricular hypertrophy). In the case of natrium-retention (diabetes, obesity, nephropathy), the treatment without diuretics is not effective. Thiazides make stronger the effects especially of ACE-inhibitors, angiotensin receptor blockers, and beta receptor blockers. The newer diuretics--with fewer side effects--have very likely extrarenal way of effects, so their long time application seems very favourable. PMID- 15125318 TI - [The human OCTN2 carnitine transporter and its mutations]. AB - The gene currently known as SLC22A5 was already sequenced in the Human Genome project, and it was annotated as the gene of the high affinity carnitine transporter (OCTN2) in 1998. After the verification of the real function of the OCTN2 several disease related mutations of the gene have been identified, albeit the entity of the primary carnitine deficiency syndrome (OMIM 212140) was separated earlier. Besides the description of the biochemical characteristics of the transporter the present review gives a summary on the mutation spectrum and the developing phenotypes according to the literature available on the Medline and to own observations. The disease spectrum includes the involvement of the cardiac muscle and the liver primarily, however, a relatively wide phenotype variability has been observed even with the same mutations. Metabolic crisis or cardiac arrest can develop in homozygotes. Since the carnitine plays a regulatory role in the mitochondrial oxidation of the long chain fatty acids, loss of OCTN2 function lead to severe impairment of the intracellular metabolism at biochemical level, which can explain the development of a severe, even life threatening condition. The cardiac symptoms can be well influenced by carnitine administration. There are indications that cardiac manifestations can develop even in heterozygotes probably in association with the gene-dose relationship; in the laboratory of the author a heterozygous C-->T transition at the 15 np of the exon V of the OCTN2 in a patient with mild cardiomyopathy and ischemic heart disease was verified. This results in a serine 280 phenylalanine (S280F) exchange affecting thereby one of the putative protein C kinase dependent phosphorylation sites. PMID- 15125319 TI - [Assessment of quality of life in functional dyspepsia. Validation of a questionnaire and its use in clinical practice]. AB - INTRODUCTION: Measurement of quality of life has gained increasing acceptance in the assessment of functional and organic gastrointestinal diseases. AIM OF THE STUDY: The purpose of study is the Hungarian adaptation and validation of a disease-specific questionnaire and the assessment of quality of life in patients with functional dyspepsia. METHODS: Validation has been performed on small groups of normal and functional dyspepsia patients. Then, one-hundred-one Helicobacter pylori positive, 98 Helicobacter pylori negative functional dyspepsia patients and 123 healthy controls have been enrolled in a prospective, controlled study. Dyspeptic symptoms were identified according to the Rome II criteria and the symptomatic subgroups (ulcer-like, dysmotility-like and alternate forms) were identified. Organic diseases were excluded by upper endoscopy and abdominal ultrasound. Helicobacter pylori infection was confirmed by the modified Giemsa stain and urease test. Healthy controls were referred by the Dimension Insurance Company and underwent gastroenterologic screening. Quality of life was assessed by the "Functional Digestive Disorders Quality of Life Questionnaire", developed by the MAPI Research Institute, Lyon, France, translated and validated in Hungarian. The effect of demographic variables on the quality of life was determined by multiple regression analysis. RESULTS: The psychometric features of the Hungarian version were similar with the original questionnaire. The standardized and transformed life quality score was of 56.3 (confidence interval: 53.3-57.7) in Helicobacter pylori positive, 60.1 (confidence interval: 58.0-62.0) in Helicobacter negative dyspeptic patients and 76.2 (confidence interval: 74.6 77.8) in healthy controls (p = 0.0001). In both dyspeptic groups life quality was significantly worse than in controls, irrespective of Helicobacter pylori status. There was no significant differences in life quality scores in subgroups of ulcer like, dysmotility-like and mixed form of dyspepsia. Only increasing age and female gender influenced the quality of life in dyspeptic patients. CONCLUSIONS: The disease-specific instrument used was able to discriminate healthy people from dyspeptic patients. Both Helicobacter pylori positive and negative patients have an impaired quality of life as compared to controls. Demographic variables have a modest influence on the quality of life of dyspeptic patients. PMID- 15125320 TI - [Unilateral Sertoli-cell androblastoma in the ovary of a young woman]. AB - The authors present a case of a Sertoli-cell type androblastoma of the ovary. The tumor is mainly seen in young women and is mostly discovered through hormonal dysfunctions linked to it. Histopathological evaluation is indispensable for an exact diagnosis. Because of its rarity, there is little experience with the tumor and the histological identification is also sometimes very challenging. The tumor is occasionally associated with Peutz-Jeghers syndrome, tumor of the thyroid, or goiter. Biologically it follows a low-malignant course with a fair prognosis, long-term complex patient follow up is necessary after unilateral oophorectomy of the diseased organ. PMID- 15125321 TI - [Treatment of interstitial lung disease]. PMID- 15125322 TI - [Treatment of diabetes insipidus]. PMID- 15125323 TI - [About Professor Tibor Verebely]. PMID- 15125324 TI - Burden of illness in patients with reflux disease--evidence from a recent comparative methodological study in Hungary. AB - BACKGROUND: The clinical and socioeconomic burden of gastro-esophageal reflux disease (GERD) is considerable. The primary symptom of GERD is heartburn, but it may also be associated with extraesophageal manifestations, such as asthma, chest pain and otolaryngologic disorders. AIM: To describe the impact of heartburn on patients' Health-Related Quality of Life (HRQL) in Hungary, using validated generic and disease-specific instruments to measure patient-reported outcomes. METHOD: Patients with symptoms of heartburn completed the Hungarian versions of the Gastrointestinal Symptom Rating Scale (GSRS), the Quality of Life in Reflux and Dyspepsia questionnaire (QOLRAD), the Short Form-36 (SF-36) and the Hospital Anxiety and Depression (HAD) scale. Frequency and severity of heartburn during the previous 7 days were also recorded. RESULTS: 136 patients completed the assessments (mean age of 47 years, SD = 13; 58% female). 53% of patients had moderate symptoms and two thirds (69%) had symptoms on 3 or more days in the previous week. Patients were most bothered by symptoms of reflux (mean GSRS score of 3.4, on a scale of 1 [not bothered] to 7 [very bothered]), abdominal pain (2.8) and indigestion (2.6). As a result of their symptoms, patients experienced problems with food and drink (mean QOLRAD score of 4.6, on a scale of 1 to 7, where 1 represents the most severe impact on daily functioning), emotional distress (4.6), impaired vitality (4.7), and sleep disturbance (4.8). This led to impaired overall HRQL across all domains (mean SF-36 score of this heartburn population compared to a general population in Hungary). Using HAD, 29% of patients were anxious and 17% were depressed. CONCLUSION: There is consistent evidence that GERD substantially impairs all aspects of health-related quality of life. PMID- 15125325 TI - Psychometric validation of the Hungarian translation of the gastrointestinal symptom rating scale (GSRS) and quality of life in reflux and dyspepsia (QOLRAD) questionnaire in patients with reflux disease. AB - BACKGROUND: Symptoms of heartburn and their impact on health-related quality of life (HRQL) are often evaluated in clinical trials. When a questionnaire is translated into a new language, a linguistic validation is necessary but not sufficient unless the psychometric characteristics have been verified. The aim of the paper is to document the psychometric characteristics of the Hungarian translation of the Gastrointestinal Symptom Rating Scale (GSRS) and Quality of Life in Reflux and Dyspepsia (QOLRAD) questionnaire. METHODS: One hundred and thirty-six patients with symptoms of heartburn (age: M = 47.3, SD = 13.4; females = 58.1%) completed the Hungarian translation of GSRS, the heartburn version of QOL-RAD, the Short-Form-36 (SF-36) and the Hospital Anxiety and Depression (HAD) scale. Eighty-seven patients were scheduled for a second visit a week later to complete the GSRS and QOLRAD again. RESULTS: The internal consistency reliability of GSRS ranged from 0.62-0.84 and of QOLRAD from 0.88-0.94, and the test-retest reliability of GSRS ranged from 0.52-0.82 and of QOLRAD from 0.68-0.82. The relevant domains of the GSRS, 'Reflux', 'Abdominal Pain' and 'Indigestion', and QOLRAD domain scores significantly correlated. All GSRS domains except 'Reflux' strongly correlated (negatively) with all SF-36 domains. All QOLRAD domains significantly correlated with all SF-36 domains. CONCLUSIONS: The psychometric characteristics of the Hungarian translations of GSRS and QOLRAD were found to be good, with satisfactory reliability and validity. The test-retest reliability of the GSRS 'Reflux' domain was, however, not optimal. PMID- 15125326 TI - [Lower esophageal sphincter pressure in patients with achalasia. Is high blood pressure frequent in these patients?]. AB - OBJECTIVE: To describe variability of lower esophageal sphincter (LES) pressure in patients with diagnosis of achalasia based in esophageal manometry. METHODS: We included 29 patients with diagnosis of achalasia confirmed by water perfusion manometry performed between July 2000 and June 2002 at the Digestive and Liver Disease Center in Guadalajara, Jalisco. Three were excluded due to impossibility of evaluating LES. RESULTS: Esophageal aperistalsis was found in all patients. LES pressure was normal in 73%, low (hypotension) in 8%, and high (hypertension) in only 19% of patients. Relaxation of LES showed different responses ranging from absence to complete relaxation after degluttion. There was absence of relaxation after degluttion in all patients with LES hypertension and hypotension. CONCLUSION: Hypertension of LES in our cohort of patients with achalasia was seen only in 19%. Knowledgement of manometric variability is fundamental to avoid errors in achalasia diagnosis and further patient treatment. PMID- 15125327 TI - [An useful classification for acute appendicitis]. AB - OBJECTIVE: To analyze clinical aspects of acute appendicitis at a teaching hospital, and to present a simple classification for acute appendicitis according to surgical findings, a classification that will indicate what postoperative treatment should be. MATERIALS AND METHODS: The study included 3,112 adult patients who had undergone surgery between September 1994 and February 2000 and who had been diagnosed preoperatively with acute appendicitis. Faced with the need to simplify description of surgical findings and to relate them to treatment, the following classification was instituted: Grade 0: No appendicitis; incidental appendectomy was carried out and antimicrobians were stopped. Grade Ia: Oedematous, ingurgitated appendix. Grade Ib: Abscessed or phlegmonous appendix, presents fibrin membranes and seropurulent liquid around appendix. Grade Ic: Necrosed appendix with no perforation. In these grades, there is very little or no presence of bacteria in periappendicular liquid or in fibrin membranes. Hence, treatment consisted of appendectomy and perioperative antimicrobians (metronidazol 500 mg and amikacine 500 mg within 2 h prior to surgery, followed by two further doses every 8 h postoperatively). Grade II: Perforated appendix with localized abscess. Treatment consisted of appendectomy and antimicrobians begun preoperatively and continuing for 3 days. A Saratoga or Penrose drain was inserted if internal tissue bed remaining after excision was raw or bloody, and might have predisposed to collection of fluid. Grade III: Complicated appendicitis with generalized peritonitis. In these cases, appendectomy was carried out with lavage to abdominal cavity. Antimicrobians were administered beginning preoperatively and were continued until patient's general condition was good, the patient had remained afebrile for 48 h and white cell count had fallen. RESULTS: Patients ranged in age from 16 to 83 years, with 62% of patients men and 38%, women. Grade I appendicitis was found to be the most common, affecting 74% of patients. Hospital stay for grade I appendicitis was 2 days; for grade II appendicitis, from 2 to 7 days; and for grade III appendicitis, from 3 to 14 days. No significant difference was noted in presence of surgical wound infection among grade I appendicitis cases, incidence being 1.1%. In grade II appendicitis cases, incidence was 17.1% and 36% in grade III cases. CONCLUSIONS: Classification of acute appendicitis as herein proposed made it possible to standardize management and treatment carried out and to predict possible complications. PMID- 15125328 TI - [Peutz-Jeghers syndrome]. AB - Peutz-Jeghers syndrome is an autonomic dominant disease characterized by hamartomatous polyps and mucocutaneous hyperpigmentation. We present 16 cases; females were more affected. The most common presenting complaints were of gastrointestinal tract. All polyps found were hamartomatous with general distribution through gastrointestinal tract. Endoscopic polypectomy should be carried out for treatment. Radiologic, endoscopic and histologic studies should be conducted for long-term follow-up, because of high risk of malignancy. PMID- 15125329 TI - [Comparison of two different vaccination schemes against Hepatitis A and B in Mexican children and adolescents]. AB - BACKGROUND: Development of multiple antigens in combined vaccines offers the advantages of reducing costs, increasing compliance and provides dual protection. Hepatitis A is an endemic disease in Mexico and hepatitis B, notwithstanding low prevalence, confers risk of progression to cirrhosis, hepatocellular carcinoma, and high medical costs in consequence. OBJECTIVE: Determine immunogenicity and reactogenicity of a combined vaccine when compared with use of conventional vaccines simultaneously. METHODS: The present study was a prospective, open, and randomized trial; 73 healthy children and adolescents were included, all with negative serologic markers. They were assigned to one of the following groups: Group 1, combined vaccine (n = 49) Twinrix (HAV 720 UE/HBV 20 micrograms), and group 2, separate vaccines (n = 24) Engerix B 20 micrograms/Havrix 720 UE. Both groups were given two-dose series at months 0 and 6. Geometric titles of antibody production (GMT) anti-HAV and anti-HBV were determined in months 1, 2, 6 and 7. Adverse reactions were registered during the study. RESULTS: No difference was observed between the two groups in age or gender. Immunogenicity anti-HAV: 100% of vaccines in both groups reached seroprotective levels (> or = 33 mUI/mL). Antibody titles in group 1 were three times higher than those in group 2 (9,696 mIU/mL vs. 3,940 mIU/mL [p = 0.003]) at the end of the study. Immunogenicity anti HBV: All subjects in both groups reached seroprotective levels (> or = 10 mIU/mL) with similar antibody titles at the end of the study (group 1: 5,603 mIU/mL vs. group 2: 5,201 mIU/mL [p = 0.55 NS]). Reactogenicity: No serious adverse reactions were observed; main were local, and frequency and characteristics were similar in both groups. CONCLUSIONS: Seroprotective levels and reactogenicity obtained from use of a combined vaccine against hepatitis A/B are acceptable when compared with use of conventional vaccines administered separately. PMID- 15125330 TI - [Cavernous hemangioma of the liver and hepatic hemangiomatosis. Indications and results of the surgical resection]. AB - INTRODUCTION: Cavernous hemangioma is the most frequent focal liver lesion. It affects mainly women and may cause symptoms such as abdominal pain, mass, and early satiety, or complications such as heart failure or coagulopathy. There are several options for treatment in symptomatic patients. However, it seems that surgical resection is the only curative treatment. AIM: Evaluate indications and results of liver resection in patients with cavernous hemangiomas and hepatic hemangiomatosis. PATIENTS AND METHODS: We conducted a retrospective analysis of clinical files of patients treated at the Instituto Nacional de Cancerologia (INCan) and the Centro Medico ISSEMYM during a 8-year period. Epidemiological data as well as diagnostic work-up and treatment were analyzed. RESULTS: From August 1995 to May 2003, 24 patients with liver hemangiomas were resected at both institutions. Twenty three were female (95.8%) and one, male (4.1%). Indications for surgery were presence of symptoms in 20 patients (83.3%), undefined diagnosis in three (12.5%), and rapid growth in one (4.1%). Most frequent symptoms were abdominal pain in 20 (83.3%) patients, followed by abdominal mass in five (20.8%), and early gastric satiety in four (16.6%). Abdominal computed tomography (CT) scan was the most frequent imaging study used in 22 patients (91.6%), followed by ultrasound in 20 (83.3%). Size of lesion ranged from 4-30 cm (X: 8.7 cm), 16 patients were submitted to formal liver resection (66.6%), and eight to enucleation (33.3%). Four patients presented operative complications (16.6%) that included postoperative bleeding in two (8.3%), fever in one (4.1%) and abdominal haematoma in one (4.1%). There was no operative mortality. Twenty two patients were asymptomatic at time of evaluation (91%). CONCLUSIONS: This lesion affects mainly women, and presence of symptoms is the most common indication for treatment. Choice of surgical procedure to be carried out depends on location and morphology of the lesion. Liver resection or enucleation are safe forms of treatment that properly controlled symptomatology. PMID- 15125331 TI - [Undifferentiated embryonal sarcoma of the liver. Case report of an adult woman]. AB - Undifferentiated (embryonal) sarcoma of the liver (USL) is a rare hepatic tumor and is generally considered an aggressive neoplasm with unfavorable prognosis. USL has a predilection for children and young adults in the first two decades of life, although a few cases have been reported in adults. The present report describes occurrence of such tumor in a 24-year-old woman who came for treatment in our hospital. She underwent exploratory laparotomy, and tumorectomy was performed. Histological examination resulted in diagnosis of USL. The patient died one month after her operation. Clinical and histopathologic features of this tumor are discussed. PMID- 15125332 TI - [Cervical esophagogastrostomy dehiscence after gastric pull-up for type I esophageal atresia. Case report of a patient successfully treated with fibrin glue and a review of the literature]. AB - INTRODUCTION: The treatment of choice for type 1 esophageal atresia is surgery to re-establish continuity of gastrointestinal tract by colon interposition or gastric pull-up. Incidence of cervical anastomosis dehiscence is 20-40%. Although it is not a serious complication, it requires a fasting period and nutritional support to achieve spontaneous closure within 2-4 weeks. AIM: To report successful treatment of cervical esophageal-gastric anastomosis dehiscence after gastric transposition using fibrin glue. CASE PRESENTATION: A 17-month-old female child, the product of a first pregnancy, was delivered by caesarean section at 35 weeks gestation in response to fetal distress; the child weighed 1,800 g at birth and had APGAR score of 6-8. At 7 h of extra-uterine life, she was diagnosed with type I esophageal atresia and cervical esophagostomy was performed. Stamm-type gastrostomy was used for enteral nutrition until she was 17 months of age. With a weight of 9 kg, the child underwent esophageal substitution with transhiatal gastric transposition, cervical anastomosis, and jejunostomy. On the seventh day postsurgery, surgical wound infection was observed, producing purulent material and saliva (45 ml in 24 h). On the 10th day, esophagogram was performed that revealed 30% anastomosis dehiscence and 2-cm fistulous tract with no abscess. On the 11th day, 3 ml of fibrinogen (80 mg/mL) activated with thrombin (1,000 uL/mL), both of human extraction (Quixil, Omrix, Tel Aviv, Israel) was applied through fistula tract. RESULT: The following day, the patient presented with cough access, expelling the fibrin glue plug. A similar dose was given that achieved a null output volume during the following 24 h. Subsequently, she began oral feeding with liquids, incorporating a normal diet over the following 2 months. At present, she is 3 years 2 months of age, weighs 15 kg, and is able to eat any kind of food without dysphagia. CONCLUSIONS: We demonstrated that use of biological adhesives can shorten time required to achieve closure of high- and low-output volume fistulas. In the present case, use of fibrin glue to obliterate the fistulous tract gave satisfactory results, reducing time to spontaneous closure and facilitating early commencement of oral feeding. Cases of enterocutaneous fistula closure reviewed in the literatura generally involve adults, but the technique can be used in children with this type of complication. PMID- 15125333 TI - [Rectal cancer and pregnancy]. AB - OBJECTIVE: To present Instituto Nacional de Cancerologia de Mexico, Gastroenterology Service management of rectal cancer in pregnant patients in a retrospective case study. CASE REPORT: Two patients were found, 25 an 34 years of age; both adenocarcinomas presented in Dukes D stage and manifested in third trimester of pregnancy. Main clinical manifestations were pelvic pain, hematochezia, and weight loss. Both patients died due to disease progression. CONCLUSIONS: Rectal cancer in pregnant patients is generally an advanced disease at diagnosis. PMID- 15125334 TI - [Choledochal cysts in the adult patient. ]. AB - INTRODUCTION: Choledochal cysts are a rare anomaly of the biliary system; only 20 30% are diagnosed in adults. The etiology remains uncertain; however, many patients with this pathology have had an anomalous pancreatobiliary junction. AIM: To evaluate recent trends regarding diagnosis and treatment of choledochal cysts. MATERIALS AND METHODS: Review of the world literature was performed on the matter of choledochal cysts. Incidence, signs and symptoms at presentation, and diagnostic tools and therapeutics are discussed. Results of these authors are also reported. RESULTS: Estimated incidence of choledochal cyst disease varies according to population studied. It is reported mainly in children; however, an increasing number of adult patients have been diagnosed with the disease. Symptoms are often unspecific and laboratory evaluation may demonstrate no variation. Ultrasonography (US) and cholangiography are both effective in defining biliary dilatation, but endoscopic retrograde colangiopancreatography (ERCP) and recently magnetic resonance cholangiopancreatography (MRCP) have been evaluated and best visualize the pancreatobiliary junction. Treatment of choledochal cyst disease is surgical and does not depend on age of patient; nonetheless, reach patient should be properly evaluated. CONCLUSIONS: Although choledochal cysts are typically diagnosed in infancy or childhood, the surgeon should not exclude the diagnosis because the patient is an adult. Cyst excision with hepaticojejunostomy is the definitive treatment of choice; nevertheless, surgical strategy should be selected based on type of cyst and the patient. PMID- 15125335 TI - Current concepts in selecting dental implant diameter size. PMID- 15125336 TI - [About values]. PMID- 15125337 TI - [States of nervousness. Useful medicinal plants]. AB - The author analyzes the effects of diverse medicinal plants such as poppy (papaver somniferum), Hawthorn, hypericum, and hops on those moderate nervous states which provoke insomnia, anxiety, or excitement as a complementary method to aid a patient overcome those states. PMID- 15125338 TI - [From project to reality. Implementation of standardized care plans]. AB - The authors describe the experience which the Apostle Santiago County Hospital in Miranda de Ebro, Burgos had when it implemented standardized treatment plans. This procedure is viewed satisfactorily and at the present time a direct follow up process continues so that the quality of those parameters examined continues to improve. PMID- 15125339 TI - [Hemolysis in blood samples. Assessment in 3 extraction systems]. AB - Blood extraction by means of a vacuum system is a technique which has been employed in our field for some time, although this method does not receive all the popularity it should. This method offers something which the classical syringe can not: greater security. Proper blood extraction leads to a decrease in problems/errors which are susceptible to appear during the pre-analysis period. The use of a system which combine the characteristics of a vacuum with those of classical syringes, such as the Holdex de Vacuette (Bio Greiner, Madrid, Espana H) can help to change this negative tendency. However, the special characteristics of this system (bendable to adapt to the site of a phlebotomy with the tube used to extract a blood sample) could cause an increase in the hemolysis in samples due to theoretically greater difficulty blood flow has in this circuit in comparison with other usual procedures in our filed (syringe and vacuum system. To test this assumption, 512 patients who had been asked to undergo a routine biochemical analysis were randomly chosen. These patients were divided into 3 groups, one for each extraction method used; 171 used Holdex de Vacuette(H), 168 used Vacutainer (V), and 173 used a syringe(J). The presence of hemolysis in these tubes was tested and the number of cases was compared to those detected among all the samples carried out during the study period. No statistically significant differences were discovered either among the 3 extraction systems used (H 2.1 or 16%, V 2.1 or 15%, J 4.2 or 23%) nor with the all samples analyzed (39, 2.02% total; 8, 1.56% study group, p < 0.001). These results let us conclude that the number of cases of hemolysis which occur when using Holdex is not greater than the number which occur when using other extraction systems and this method can be a good alternative to the conventional vacuum system. PMID- 15125340 TI - [Childhood obesity. Are prevention and control programs efficient? A study based on original articles]. AB - Continuing with the importance given throughout the world to the prevalence of adults and children who are overweight or obese. The authors review the effectiveness of programs for the prevention and control of childhood obesity by analysing original articles in this second section of their article. A complete bibliography accompanies this article. PMID- 15125341 TI - [The power of values. A question of professionalism]. AB - In order to include the term on which my thoughts shall turn, related to professionalism, I strive to find the words which project a specific sense to the concrete aspect of what I want to express integrating values inherently when I say profession. My point of reference is Stephen. Toulmin, an interesting English philosopher and disciple of Ludwig Wittgenstein, who in his last known publication develops his thoughts about rationality and coins the term reasonability, which he defines as the capacity to be sensitive to the thousands of ways the same situation can be presented and he points out its meaning with these words: "One hopes that the human values of reasonability are self-justified in rationality". Following this same line of thinking, I propose that the human values of professionalism be self-justified by the professionalism nurses have. Based on this reflection, I open for debate the concept of PROFESSIONALISM, hoping to arrive beyond the norms which can determine the future professional, since these shall always be limited by one's own personal experiences and by the review of our short bibliography on this issue. PMID- 15125342 TI - [Keeping company in an emotional trip. Emotional intelligence applied to the help relationship]. AB - In order to be a good professional and caretaker, it is essential to work on one's capacity to manage one's own feelings and emotions in an adaptable, intelligent manner. This set of abilities form part of the concept known as Emotional Intelligence. One can only give to another what one is and one knows how to give to oneself. The five ability groups which make up affective or emotional intelligence are: self-knowledge, self-control, self-motivation, empathy and relationship abilities. All are necessary in order to carry out good management of our feelings and emotions. PMID- 15125343 TI - [Prevention and treatment of pressure ulcers]. PMID- 15125344 TI - [Clinical pathways. Time recovery and improvement of quality of care]. AB - The authors describe the real experience the Holy Spirit Hospital had regarding the start of clinical trajectories. The authors highlight the numerous advantages these provide for professionals and patients. PMID- 15125345 TI - [Which contraceptives do we use? Current trends]. AB - This article is a descriptive study about two variables, age and anti-conceptive method used, over two consecutive time periods, 2000 and 2001, made on women who made use of a Family Planning Service located at a Basic Health Service Unit in Gava, Barcelona. Condoms, used by 43.49%, were the anti-conceptive method most frequently used by adolescent women in 2000 while in 2001, oral anti-conceptive methods, used by 44.68%, were the most frequently used method. Among adult women, there were no differences either year; oral anti-conceptives were used by 27%, and condoms were used by 24%. Oral anti-conceptives are the most frequently used method among women in both age groups. Among adolescents, this study showed how oral anti-conceptive use has replaced the method most highly recommended for this age group, condoms. PMID- 15125346 TI - [Piercings. A new demand for nursing]. AB - The new legal regulations and agreements dealing with the type of locales and personnel authorized to implant body rings through pierced holes known popularly as piercings brings with it the need for nurses to learn about this practice and to adapt our habitual sanitary techniques, in an urgent manner, in order to cover the increasing aesthetic demand. The nursing profession needs to amplify its range of treatments for this piercing practice so it is not limited to merely inserting rings or studs in holes. PMID- 15125347 TI - [Women: reality and desire. Thoughts on the International Women's Day]. PMID- 15125348 TI - [Already in the 21st Century... new challenges in health sciences]. AB - In the words of the author, the 21st Century shall "witness the consolidation of the change from a society which cures to a society which cares for patients". This means that the nursing profession will assume a series of challenges, known as the management challenge, new missions for the nursing professional and the challenge to provide excellence in patient treatment. Probably only with attitudes and reflexive practices, with teamwork and continuous professional development which lasts through one's life, we will provide the correct response. PMID- 15125349 TI - [Silent victims]. PMID- 15125350 TI - [Treatment of tobacco dependence: brief and intensive clinical intervention]. AB - This article presents a literature review on the different methods of tobacco cessation that proved their efficiency through the recent meta-analyses. Smoking help cessation can consist in a systematic minimal advice that must be achieved physician or other professional of health throughout during their contact with their patients. This minimal intervention permits to get 5% of tobacco abstinence. The intensive clinical intervention is a very useful to higher dependent smokers or presenting anxiety and depressive disorders. It consists in a psychological combined approach by a behavioral and cognitive therapy and a pharmacological approach with two types of treatment: nicotine replacement therapy and the bupropion, the other "methods" not having proven their efficiency. The success rate of this intervention depends in the time of contact and the number of session. In order to promote tobacco cessation, it is necessary to widen the pharmacological product range and to educate health professionals in smoking cessation. PMID- 15125351 TI - [Mechanical ventilation in intensive care units: indications, modalities and complications. Results of a prospective multicenter survey in Tunisia]. AB - The survey was performed during the month of March 1998 and concerned 9 ICUs located in teaching hospitals. To be included each ICU had to MV for more than 12 hours were included in the study and had a 28 day follow-up in the ICU or until hospital discharge. Collected parameters were indications of MV, modalities of MV and of weaning, complication and outcome at hospital discharge. Assist-control ventilation was the most used ventilation modality (69.8%). Weaning of MV was performed in 63% of the study patients and was based on a once-a-day attempt of spontaneous breathing through a T-piece (59.5%) and a combination of intermittent mandatory ventilation with pressure support (IMV-PS: 27%) or pressure support alone (11.2%). Mean length of hospital stay was 19.7 +/- 15.9 days of which 11.6 days were spent in the ICU. Fifty nine patients (54%) were alive at discharge form the ICU of whom 4 ultimately died during their hospital stay. MV practice as well as ICU facilities are not homogenous in Tunisia. Recommendations and guidelines should be built in order to standardize MV practice in Tunisia. PMID- 15125352 TI - [Comparison of three protocols in the control of cardiovascular response to suspended laryngoscopy in ENT surgery]. AB - The objective was to compare the effect of three pharmacological agents on the hemodynamic response to suspended laryngoscopy in micro ENT Surgery. Double blind randomised prospective study having included: groupe A (n = 16) having benefited before laryngoscopy of 150 mcg/kg esmolol, groupe B (n = 16) 15 mcg/kg of nicardipine, groupe C (n = 15) 1 mg/kg of lidocaine and groupe D (n = 16), placebo group. All groups were comparable for demographic and anesthetic data. A significant reduction of the heart rate and pressure rate product were noted in esmolol group during the endoscopic act and maintained until arousal. Whereas there was no difference in the blood pressure during the procedure whatever the pharmacological agent for prevention of cardiovascular complications for patients to risk in micro ENT surgery. PMID- 15125353 TI - [Surgical treatment of Basedow's disease. Retrospective study of 40 cases]. AB - Graves' disease is an autoimmune disorder. Its diagnosis is easy. Its treatment is delicate in lack of etiologic treatment. We report a retrospective study of 40 cases operated in nine years period. The failure of the medical treatment constituted the main operative indication in our series (57.5%). A subtotal thyroidectomy have been accomplished in 72.5% and total thyroidectomy in 22.5% of cases. We noted one case of definitive hypoparathyroidism after surgery. Our endocrinal results were characterised by the occurrence of hyperthyroidism in 2 cases after subtotal thyroidectomy. For many authors, surgery is the best treatment of graves' disorder and the actual tendency is to realise a total thyroidectomy. Our results encourage us to adhere to the therapeutic method. PMID- 15125354 TI - [non-Q wave myocardial infarction: study of 31 cases]. AB - The frequency of non-Q wave myocardial infarction is permanently increasing. In this retrospective study, we have tried to establish the clinical features, the prognosis and the therapeutic possibilities in this entity. We have studied the clinical history, the physical examination data, the results of the different explorations and the short and long term general course in 31 patients with a non Q wave myocardial infarction. The main feature of this acute coronary syndrome is the preservation of the myocardial function (normal in 64% of the patients). Concerning the angiographic finding, we note a high prevalence of severe coronary damages particularly of the stenosis of the left main coronary artery (13.5%); but also an important proportion of normal coronary angiographies (20%). In addition to the known factors associated with a poor prognosis in coronary artery disease, we insist on the severity of the initial depression of the ST-segment. The prognosis of the non Q wave myocardial infarction is better at the initial phase than that of the "transmural infarction". However, it becomes similar or worse at the long term general course. The important progress in the prognosis markers and in the anti-thrombotic and interventional therapies may maintain at the long term course the good initial prognosis. PMID- 15125355 TI - [The obstetrical advantages of epidural analgesia in a trial of labor]. AB - Two series of labour trial were compared in a forward-looking study. The first series of 24 parturient women (series I) to whom the labour trial took place under peridural analgesy. The second series of 80 parturient women (series II) to whom the labour trial took place without peridural analgesy. The rate of caesarean section is significantly less important in the series I (33.3% versus 58%) P = 0.37. The average duration of the labour trial is significantly prolonged under peridural analgesy but without bared effects for the newborn children. The authors consider that the peridural analgesy is the method that brings ideal conditions for a real test allowing to eliminate dynamic dystocia and maternal restlessness in order to have an accurate cephalo-pelvic confrontation. PMID- 15125356 TI - [Intussusception in adults. Thirteen case reports]. AB - This study was retrospective. The authors analyse the clinical, etiological and therapeutic aspects of the intestinal intussusception based on 13 adults observed over 14 years in the surgical department of the university hospital in Monastir. This disease was rare because accounted for 2.6% of all cases of intestinal obstruction. The diagnosis was made in the majority of cases during the operation (8/13). Abdominal pain was noted in all cases. The underlying pathologic processes were identified in 69%. Operation was required in every case. During the surgery, the lead point was identified in the small bowel in 12 cases and in the colon in one case. An intestinal tumor was found in four patients (39%), only one of which was malignant. Adult intussusception is an unusual cause of abdominal pain and bowel obstruction. It requires preoperative diagnosis. Operative management is always necessary because this condition is almost always secondary to definable lesion. PMID- 15125357 TI - [Mesial temporal lobe epilepsy. Nine case reports]. AB - INTRODUCTION: Mesiotemporal epilepsy (MTLE) is a clinical syndrome characterised by the association of a history of febrile seizures, a homogenous clinical presentation of seizures, temporal interictal and ictal EEG recordings and an underlying pathology that is mesial sclerosis. MTLE is the most common type of medically intractable partial epilepsy with a drug-resistance in 90% of cases. OBJECT: The aim of this study is to describe the clinical, EEG and MRI findings of 9 patients with MTLE attending the outpatient clinic of Charles Nicolle Hospital. RESULTS: The median age of our study population was 30 years. A history of febrile seizures was found in 5 patients. Hippocampal atrophy was found in all the cases right in 4 cases and left in 5 cases. Drug-resistance was observed in 7 patients. No patient underwent surgery. CONCLUSION: It is important in front of medically intractable partial epilepsy to evoke MTLE, to confirm the diagnosis with neuro-imaging and to propose an interdisciplinary therapeutic approach including neurologists, epileptologists and neurosurgeons. PMID- 15125358 TI - [Cervical-facial skin tuberculosis. Three case reports]. AB - The cervical-face skin tuberculosis is fairly uncommon pathology, it is caused by the Tuberculosis mycobacterium (or Bacille de Koch). With human or bovine origin this pathology is actually increased as like as AIDS in countries where it was eradicated. Skin lesion may be primitive or secondary to another pre-existing tuberculosis lesions, it may be have many clinical appearance with the same characteristics, as knew as, long evolution and anesthetics sequels. We report in this paper three cases of cervical-face tuberculosis with different age, the diagnosis was given by biology and histology and treated medically. PMID- 15125359 TI - [Multiple myeloma and HIV/AIDS infection. Three case reports]. AB - HIV/AIDS infection rages at the endemic state in Sub Saharan African and especially in our country. We are reporting 3 observations of patients, all of females affected by HIV/AIDS and multiple's myeloma. They are all in-patients in the department of haematology, University hospital of Brazzaville, from 2000 to 2002. In two cases out of three, multiple's myeloma is discovered after the tracking of HIV/AIDS infection. In the other case, the tracking HIV/AIDS seems to be posterior to the multiple's myeloma diagnostisis. HIV/AIDS is symptomatic in the tree cases and under antiviral treatment. Multiple's Myeloma is diagnosed at an advanced stage. It is about IgG myeloma in two cases and IgA myeloma in the other. The rate of the T4 lymphocytes is noted spontaneously to 204 and 486 by mm3, and 390 by mm3 in a patient under antiviral treatment before her hospitalisation. The measure of the viral intensity is not achieved. The poly chemotherapy of type VMCP and VAMCP is driven without major complications and under anti-infectious prophylaxis. The receding is still insufficient to affirm the medium-term evolution and to determine the prognosis of the malign blood disease. The description of these three observations confirms the implication of the human retrovirus and in particular of the VIH/SIDA in the lymphoma genesis. PMID- 15125360 TI - [Splenic abscess: from diagnosis to treatment]. AB - Splenic abscess are a rare and severe pathology causing difficulty in diagnosis and treatment. The Clinical presentation usually associated with abdominal pain in the left upper quadrant, fever and splenomegaly. Diagnosis will be confirmed by morphologic examination such as ultra sonography and computed tomography. Percutaneous drainage procedure made in first intention can be used either for curative purpose or to prepare patient for surgery. We report four cases of splenic abscess collected in our departement between 1997 and 2000 in which percutaneous drainage procedure was trial first. Two of them were successful and the others were completed by splenectomy. PMID- 15125361 TI - [An unusual cause of secondary scoliosis: vertebral osteoblastoma]. AB - We report the observation of a 22-year-old man suffering from nocturnal back pain for 2 years. The clinical examination shows a scoliosis. Imaging suggests either malignant tumor or an aggressive osteoblastoma of the sixth thoracic vertebra. After radical removal of the tumor, histopathologic examination confirms the diagnosis of osteoblastoma. Like any other neoplasma, osteoblastoma should be detected and removed early, before being responsible for non-reversing neurological complications. PMID- 15125362 TI - Trauma of an ectopic kidney. A case report. AB - Ectopic kidney is a rare pathology. It is usually misknown and often revealed by a complication. We report a case of a 30-year-old male patient who presented in emergency with a right thoraco-abdominal trauma and a benign head trauma. Injury evaluation revealed a right ectopic pelvic kidney with a grade IV laceration. Furthermore, there was a hepatic contusion in the sixth segment and a fracture of the ninth, tenth and eleventh right ribs. Therapeutic attitude consisted on a successful conservative and nonoperative treatment for both lesions. CT scan is of utmost importance in diagnosis. Management is the same as for normally positioned kidney. PMID- 15125364 TI - [Adefovir dipivoxil--a further step in the control of chronic viral hepatitis B. There is still a long way to go]. PMID- 15125365 TI - [Iron, free radicals and atherosclerosis]. PMID- 15125366 TI - [Is there a relation between serum ferritin levels, oxidized LDL antibodies and vitamins and risk for coronary atherosclerosis]. PMID- 15125367 TI - [Percutaneous revascularization and elevation of troponin]. PMID- 15125368 TI - [Recommendations for diagnosis and therapy of chronic heart failure 2001-2003]. PMID- 15125369 TI - [Ferritin, oxidative stress and coronary atherosclerosis]. AB - BACKGROUND AND AIM: In the recent years several studies showed the association between body iron stores, represented by serum ferritin, and atherosclerosis. It was proposed that iron bound to ferritin catalyzes the formation of highly reactive forms of oxygen free radicals which subsequently cause the oxidative modification of atherogenic lipoproteins. Aim of our study was to compare serum ferritin concentrations and certain markers of oxidative stress in patients with and without coronarographically assessed coronary vascular disease. METHODS AND RESULTS: Measurements were performed in 216 subjects at the age of 35-60 years. The patient group included 76 patients with coronarographically assessed coronary vascular disease (CVD) (mean age 51.16 +/- 5.713 years) and 140 healthy controls (mean age 50.21 +/- 5.331 years). The plasma concentration of ferritin was higher in patients (169.04 +/- 63.899 micrograms/l) than controls (87.70 +/- 41.394 micrograms/l), p < 0.001. The group of patients revealed significantly lower plasma concentrations of anti-oxLDL antibodies, nitrites/nitrates, tocopherol and high density lipoprotein cholesterol (HDL-cholesterol) than controls; on the contrary patients had significantly higher concentrations of hemoglobin, thrombocytes and triacylglycerols. In the whole cohort of investigated subjects, ferritin correlated positively with retinol, body mass index (BMI), total cholesterol, triacylglycerols, low density lipoprotein cholesterol (LDL cholesterol), blood glucose, creatinine, uric acid, alaninaminotransferase (ALT), aspartateaminotransferase (AST), hematocrite, erythrocytes, with occurrence of CVD and with sex. Inverse correlation was observed between ferritin and HDL cholesterol. CONCLUSIONS: Our observations are consistent with the hypothesis that high stored iron levels, measured by serum ferritin concentrations, may contribute to the oxidative stress and thus elevate the risk for development of CVD. PMID- 15125370 TI - [Increase in troponin levels after coronary angioplasty (PTCA) in stable angina pectoris]. AB - GOAL: To determine frequency of elevated troponin levels following PTCA in patients with stable angina pectoris. To identify risk factors related to troponin elevation. METHOD: Multicentric prospective study. Troponin I level (cTnI) was determined in a group of 261 patients treated for stable angina pectoris with coronary angioplasty (PTCA) 12 hours after the intervention. A group of patients with cTnI levels above the upper level of a normal range was compared to patients without troponin elevation. Clinical, angiography, and peri procedural indicators were assessed and frequency of their incidence in both groups of patients was compared. RESULTS: Elevation of cTnI levels above the upper levels of the normal range was identified in 32 patients (12.3%). There were no differences in age, risk factors for ischemic heart disease (IHD), nor number of impaired coronary arteries between this group of patients and the rest of them. Associated antithrombotic treatment (acetylsalicylic acid + ticlopidine 87.5% vs. 86.9%, p = NS; low-molecular heparin for PTCA 46.9% vs. 57.2%, p = NS) was comparable in both groups. On angiography, according to ACC/AHA, lesions were worse in patients with elevated cTnI (2.73 vs. 2.33, p = 0.02). Troponin elevation was significantly more often connected with calcification of coronary arteries (37.5% vs. 17%, p = 0.03), with intracoronary thrombus on angiography (15.6% vs. 2.2%, p = 0.05), and with increased number of implanted stents (1.13 vs. 0.90, p = 0.03). Incidence of peri-procedural complications (temporarily occluded artery, arterial dissection type C and worse, forced administration of inhibitors GP IIb/IIIa) was comparable. Chest pain after PTCA was accompanied with consecutive elevation of cTnI in 40%, while in absence of chest pain cTnI was elevated only in 8% of patients. CONCLUSION: Elevation of troponin after PTCA in stable angina pectoris is significantly related to angiography findings in treated lesion. Elevation of cTnI is comparable both in use of unfractionated heparin during PTCA and in use of low-molecular heparin during PTCA. A combined antiaggregation treatment with acetyl salicylic acid (ASA) and ticlopidine did not lead to a lower incidence of cTnI elevation compared to treatment only with ASA. Heaviness in chest after PTCA has low positive and high negative predictive value for cTnI elevation. PMID- 15125371 TI - [Can serologic markers be indicators of dietary errors in patients with celiac disease?]. AB - Coeliac disease is a disease of small intestine requiring life-long strict gluten free diet to avoid acute and chronic complications. To reach maximum adherence to the diet is in some of coeliac patients, especially adults, difficult because it requires distinct changes in eating habits. To diagnose coeliac disease tests of antigliadin and anti-endomysial antibodies in serum are used. Authors monitored levels of antibodies in 32 adults 3, 6, and 12 month after histology validation of coeliac disease and after recommendation of a gluten free diet. The results were compared with data from nutritional history. Maximum adherence to the diet indicated 24 patients (75%), occasional consummation of gluten indicated 6 patients (19%) and more frequent breaking a diet indicated 2 patients (6%). In a group which adhered to the diet the most rapid was a decline in levels of anti endomysial antibodies, less rapid was a decline in levels of IgA-class antigliadin antibodies, and the least rapid was a decline in levels of IgG-class antigliadin antibodies. In the group with occasional intake of gluten was the decline slower and in the group with frequent dietary mistakes levels of antibodies have not declined at all. Adherence to the diet positively correlated with level of accomplished education of patients. Monitoring of titter kinetics proved to be a good indicator of discipline and cooperation of patients during treatment with gluten free diet. PMID- 15125372 TI - [Causes of hospitalization in patients on chronic hemodialysis]. AB - Patients taking dialysis regularly form a group with higher morbidity and mortality compared with common population. The risk factors of the hospitalization in future in these patients are supposed to be: older age, history of cardiovascular disease, comorbidity, vascular access other than arterio-venous fistula, certain types of nephropathy and serum albumin level < 30 g/l. The number of patients in chronic dialysis treatment in Slovakia rises. Therefore we have performed a retrospective study. It's aim was to evaluate the main reasons and risk factors of hospitalizations in chronic haemodialysis patients in Turciansky region. METHODS: 80 patients undergoing regular haemodialysis treatment in 2 dialysis centres during 24 months were included. Following data were collected: age, gender, comorbidity, type of nephropathy, residual diuresis, some data connected with dialysis treatment, laboratory parametres and body mass index. RESULTS: During the given period of time 66 per cent of the patients of our sample required hospitalization. The main reasons of their hospitalization were complications of vascular access (13%), surgery (12%), the sepsis (9%) and serious bleeding (9%). Hospitalized patients showed significantly lower BMI and residual diuresis compared with non-hospitalized ones. They also suffered from greater amount of other diseases. As for gender prevailed men and patients with a history of cardiovascular disease, thrombosis and peptic ulcer. There was no connection between morbidity and age, type of vascular access and laboratory parameters observed. PMID- 15125373 TI - [Treatment of chronic postirradiation proctitis with argon plasma coagulation ]. AB - Chronic radiation proctitis calls for a disease of the rectum due to irradiation of pelvic malignancies. The results of treatment of 8 patients with hematochezia from chronic radiation proctitis are presented here. They were treated with argon plazmakoagulation during 1-4 sessions. Hematochezia has disappeared in all cases. Rectal pain was the reason for hospitalization in one case. After the treatment, improved quality of live was claimed by all patients. PMID- 15125374 TI - [Adefovir dipivoxil is a new drug for treatment of chronic hepatitis B]. AB - Adefovir dipivoxil is a new nucleotide analog derived from adenosin monophosphate. At the 10 mg/day dose significantly decreases HBV DNA in serum, activity of transaminases, and leads to better histology results in liver assessment of adults with hepatitis B in the phase of active viral replication. Treatment with Adefovir dipivoxil is a long term treatment (48 weeks), it is well tolerated, and side effects are moderate and reversible. It can be administered to HBeAg positive patients, to patients with HBV mutant unable to create HBeAg if positive HBV DNA in serum, and to patients with lamivudine resistant HBV mutant. PMID- 15125375 TI - [Diagnosis and therapy of chronic heart failure. European Cardiology Society Commission for Preparation of Practical Recommendations and the Conference on Improving the Quality of Clinical Practice and Patient Care in Europe]. PMID- 15125376 TI - [Recommendations for diagnosis and therapy of chronic heart failure 2001--the short version]. PMID- 15125377 TI - [Acute aortic dissection in a female patient with Marfan syndrome]. AB - The group of patients with the Marfan syndrome is used to suffer from aneurysm and dilatation of the ascendent aorta, therefore the reason echocardiography examination is necessary. But in spite of this, although the dimensions (measures) are stable, we do really not know when the dissection appeared. This complication is the most usual cause of death these patients. But if we recognize the diagnosis in time and if we treat it correctly, this complication can be treated with the great difference in mortality. That is while we must think of this possible complication in case of atypical breast ache. The following case describes the acute dissection of the aorta in the proximal and also descendent part together with the acute aortal significant regurgitation and the volume overload of the left chamber appeared on female--patient with the Marfan syndrome. PMID- 15125378 TI - [Large-droplet liver steatosis in celiac disease]. AB - Authors monitored a case of a 25 years old woman who was admitted for swelling of lower limbs. Laboratory results showed hypoproteinemia, elevation of liver enzymes, and prolonged prothrombin time. Ultrasound examination proved hepatomegalia with diffusely hyperechogenic liver without central lesion. Computer tomography confirmed hepatomegalia with diffusely hyperechogenic liver and a suspicion of liver steatosis was expressed. Liver biopsy confirmed serious diffuse large droplet steatosis of unclear genesis. Carried out examinations excluded infectious and autoimmune liver diseases, metabolic diseases, and congenital liver diseases (Wilson's disease, porphyria, haemochromatosis etc.). Laboratory results showed gliadin, endomysin, and reticulin antibodies. An enteroscopy picture showed villi decrease. Histology examination of a biopsy specimen confirmed total villi atrophy with non-differentiated enterocytes and round-cell cellulisation of epithelium and proprium. Histology and histochemical findings were distinct proves of coeliac disease. A patient was prescribed a gluten free diet. Her metabolic parameters (normalisation of albumine levels, prothrombine time, and trace elements) and anino transferase levels gradually improved. This case documents development of a serious liver disorder as a result of malnutrition which developed in a young woman as a result of unrecognised coeliac disease. PMID- 15125379 TI - Correlation between myocardial uptake of technetium-99m-sestamibi and pressure derived myocardial fractional flow reserve. AB - OBJECTIVES: Development of the coronary pressure wire has facilitated the measurement of fractional flow reserve (FFR) to assess the functional severity of coronary artery stenoses. METHODS: This study evaluated the correlations between FFR and myocardial direct counts of technetium-99m(99mTc)-sestamibi in 20 patients (16 men, 4 women, mean age 66 +/- 8 years) who underwent 99mTc-sestamibi single-photon emission computed tomography (SPECT) with the 2-day protocol using 740 MBq of 99mTc-sestamibi each day. Visual assessment of myocardial imaging and quantitative analysis with the measurement of percent uptake and direct count of 99mTc-sestamibi were performed. RESULTS: Visual assessment of myocardial imaging revealed that reversibility of 99mTc-sestamibi perfusion defects was correlated with FFR of < 0.75, which is regarded as functionally important stenosis (17/20 vs 3/20, kappa = 0.71, p < 0.002). Regional reversibility score did not correlate with FFR (r = -0.40, p = NS). Quantitative analysis revealed that the change in 99mTc-sestamibi percent uptake with pharmacologic stress using adenosine triphosphate disodium (ATP) also did not correlate with FFR (r = 0.35, p = NS). In contrast, percent increase in 99mTc direct counts with ATP was lower in patients with FFR of < 0.75 than in those with FFR of > = 0.75 (-4 +/- 16% vs 24 +/- 30%, p < 0.01). In addition, a significant correlation (r = 0.70, p < 0.001) was observed between percent increase in 99mTc direct counts with ATP and FFR. CONCLUSIONS: These results suggest that quantitative analysis of 99mTc-sestamibi scintigraphy enables the assessment of the magnitude of functional significance of coronary stenosis. PMID- 15125380 TI - [Relationship between lung-to-heart uptake ratio of technetium-99m-tetrofosmin during exercise myocardial single photon emission computed tomographic imaging and the number of diseased coronary arteries in patients with effort angina pectoris without myocardial infarction]. AB - OBJECTIVES: Increased lung uptake of thallium-201 in exercise myocardial perfusion imaging is a reliable marker of multivessel disease in patients with ischemic heart disease. This study investigated whether the lung-to-heart uptake ratio with techenetium-99m(99mTc)-tetrofosmin also provides valuable information to detect patients with multivessel disease. METHODS: Fifty-three consecutive patients (35 men, 18 women, mean age 66 +/- 11 years; single-vessel disease: 29, double-vessel disease: 16, triple-vessel disease: 8) with stable effort angina pectoris without prior myocardial infarction and 17 control subjects (12 men, 5 women, mean age 62 +/- 9 years) underwent exercise myocardial perfusion imaging with 99mTc-tetrofosmin and coronary angiography in January 2000 to December 2002. The lung-to-heart uptake ratio was calculated on an anterior projection before reconstruction of the exercise single photon emission computed tomographic images. RESULTS: The mean lung-to-heart uptake ratio was 0.34 +/- 0.04, 0.38 +/- 0.07, 0.41 +/- 0.05, and 0.46 +/- 0.09, in patients with normal coronary, single vessel disease, double-vessel disease, and triple-vessel disease, respectively. Significantly higher lung-to-heart uptake ratio was associated with more diseased vessels (p < 0.05). Multivessel disease could be detected with a sensitivity of 67% and a specificity of 74% if the cut-off point of the lung-to-heart uptake ratio was set as 0.4. Combining lung-to-heart uptake ratio with conventional myocardial perfusion imaging improved the sensitivity to detect multivessel disease to 83% and the specificity to 74%. CONCLUSIONS: Lung-to-heart uptake ratio measured by exercise myocardial scintigraphy with 99mTc-tetrofosmin can provide clinically useful information to detect multivessel disease in patients with ischemic heart disease. PMID- 15125381 TI - [Mechanisms of impaired coronary flow reserve in patients with aortic stenosis: transthoracic Doppler echocardiographic study]. AB - OBJECTIVES: Development of left ventricular hypertrophy in severe aortic stenosis is associated with coronary microcirculatory dysfunction, as demonstrated by impaired coronary flow reserve. Recently, coronary flow reserve can be assessed noninvasively by transthoracic Doppler echocardiography (TTDE). This study assessed the relationship between coronary flow reserve obtained by TTDE and the hemodynamic parameters and left ventricular mass index in patients with aortic stenosis. METHODS: Consecutive 29 patients (15 men, 14 women, mean age 72 +/- 11 years) with isolated mild to severe aortic stenosis were studied using TTDE to assess coronary flow reserve. Peak transvalvular pressure gradient across the aortic valve (peak AVG) and aortic valve area were measured by TTDE. Left ventricular mass index was measured by echocardiography. RESULTS: There were significant correlations between coronary flow reserve and peak AVG (r = -0.570, p = 0.001), left ventricular mass index (r = -0.620, p < 0.001), aortic valve area (r = 0.740, p < 0.001), and left ventricular rate pressure product (r = 0.660, p < 0.001). Multiple regression analysis showed that aortic valve area and peak AVG were independent factors for coronary flow reserve (p < 0.001, p = 0.048). CONCLUSIONS: Impairment of coronary flow reserve in patients with aortic stenosis is related to aortic valve area and peak AVG, rather than the degree of left ventricular hypertrophy. PMID- 15125382 TI - [Therapeutic angiogenesis by autologous transplantation of bone-marrow cells in a patient with progressive limb ischemia due to arteriosclerosis obliterans: a case report]. AB - A 78-year-old woman with arteriosclerosis obliterans was admitted with complaints of progressive ischemic change and resting pain of the left leg. She had severe ischemic limb in stage IV of the Fontaine classification. Angiography showed total occlusion of the three main arteries with poor collateral flow at the left crus. Autologous transplantation of bone-marrow mononuclear cell to her left leg was performed to achieve therapeutic angiogenesis. One month later, follow-up angiography showed excellent angiogenesis of collateral vessels at the left crus. Her pain was relieved and leg amputation was not necessary. Recently, the TACT study in Japan showed the efficacy of angiogenesis by autologous bone-marrow transplantation for peripheral artery disease. This case illustrates the efficacy of this therapy. PMID- 15125383 TI - [Fulminant myocarditis treated with percutaneous cardiopulmonary support and long term complications: three case reports]. AB - A 16-year-old female underwent percutaneous cardiopulmonary support (PCPS) for treatment-resistant ventricular tachycardia, but she could not be weaned from PCPS early without complications. A 44-year-old female underwent PCPS for low cardiac output syndrome with mainly heparin used for anticoagulation. With long term PCPS, the activated clotting time became unstable, and she died due to fatal hemorrhagic complications in the acute stage. A 71-year-old female underwent PCPS for low cardiac output syndrome with mainly nafamostat mesilate used for anticoagulation. Despite long-term extracorporeal circulation, she was weaned from PCPS without hemorrhagic complications. However, she died of multiple organ failure and systemic cytomegalovirus infection in the chronic stage. Myocardial recovery was delayed in Cases 2 and 3, so long-term PCPS was required, which resulted in severe complications. To prevent hemorrhagic complications, nafamostat mesilate should be given and activated clotting time should be measured frequently. To prevent multiple organ failure, the appropriate initial PCPS flow should be established after the evaluation of urinary output, saturation of venous oxygen, and splanchnic circulation such as arterial ketone body ratio and gastric acid secretion. PMID- 15125384 TI - [Cystic mass in the left ventricular outflow tract in a 76-year-old man]. PMID- 15125385 TI - [Guidelines for diagnosis and management of cardiovascular sequelae in Kawasaki disease (JCS 2003)]. PMID- 15125386 TI - [Biological adequacy--what does it mean?]. AB - Appropriate initiation of dialysis is of an outstanding importance in the treatment of patients with end-stage renal disease. It prevents development of irreversible uremic complication and enables selection of the most appropriate dialysis modality for the individual patient. The major causes of morbidity and mortality in dialysis patients are cardiovascular diseases. Hypertension and hyperlipidemia are commonly found in dialysis patients as well as anemia, chronic inflammation and fluid overload, all of which are found to be risk factors for the development of cardiovascular diseases. Arterial hypertension is the main risk factor for left ventricular hypertrophy, and there is clear evidence that control of hypertension has a beneficial effect on left ventricular hypertrophy. It is best achieved by correction of overhydration and maintenance of dry weight. Modern dialysis machines are capable of changing electrolyte concentrations, which reduces intradialytic cardiovascular complications, incidence of cardiac arrhythmias and hypotension. Correction of anemia with erythropoietin results in regression of left ventricular hypertrophy and improvement of the quality of life and defense against microorganisms. Chronic inflammation can be prevented with the use of biocompatible high-flux dialysis membranes and sterile dialysate, which are also important for the prevention of oxidative stress involved in the increase of LDL oxygenation and incorporation into the intimal layer of the vessels. Low molecular weight heparins by their action on lipoprotein lipase serve as an additional factor that suppresses development of atherosclerotic plaque in dialysis patients. Optimal dialysis dose decreases the mortality and morbidity rates. High-flux membranes or prolongation of dialysis session are modalities for dialysis dose improvement. Individualized approach to preparation of dialysis solutions has resulted in better control of fluid overload and intradialytic hyper- or hypotension, reduction in the incidence of arrhythmias, improvement of hemodynamic stability, and delay of renal osteodystrophy. Malnutrition is a relatively common problem in dialysis patients that may be secondary to poor nutritional intake, inadequate amount of dialysis, lack of appetite, acidosis, associated disease, and/or increase in protein catabolism. The most appropriate approach includes individualization of dietary prescription according to the nutritionist's advice, increase of dialysis dose with biocompatible membranes, and use of sterile bicarbonate dialysate with glucose and erythropoietin. The major goal of adequate dialysis is not just improvement in survival of dialysis patients, but also improvement in the quality of their lives. PMID- 15125387 TI - [Bone histology in postmenopausal osteoporosis--variations in cellular activity]. AB - AIM: Modern understanding of the etiology of postmenopausal osteoporosis is based on the imbalance between bone resorption and formation due to estrogen deficiency, which may take several forms and combinations of decreased and/or increased activity of both or one cell type. Studies of postmenopausal osteoporosis have pointed to the existence of heterogeneity in the remodeling imbalance. Bone histology analyzed in a group of women with established postmenopausal osteoporosis undergoing bone biopsy is part of the diagnostic procedure. Data were compared and grouped according to the published histomorphometric classification of postmenopausal osteoporosis. METHODS: The study included 43 postmenopausal women aged 44-71 years with osteoporosis established by densitometry of the lumbar spine and hip. Secondary causes of osteoporosis were ruled out. Full thickness transiliacai bone biopsy specimens were obtained after double labeling regime with oxytetracycline (Geomycin, Pliva). Biopsy specimens were processed for undecalcified embedding in resin and sections stained by Goldner trichrome and toluidine blue, or used for fluorescence microscopy. A grid attached to the microscope eyepiece was used for histomorphometry. The following parameters were assessed according to the recommendations of the American Society for Bone and Mineral Research: bone volume (BV/TV, %), osteoid surface (OS/BS, %), osteoblast surface (Ob. S/BS, %), osteoid volume (OV/BV, %), osteoid thickness (O. Th, m), osteoclast surface (Oc. S/BS, %), mineral apposition rate (MAR, m/day). Thus obtained data were compared to published reference data for normal healthy population and also expressed as z scores (the number of standard deviations by which the value differs from the mean of the normal age and sex matched controls). The study was approved by the hospital ethics committee. All patients signed an informed consent to take part in the clinical study. DISCUSSION: Histomorphometric analysis of bone biopsy demonstrated that on an average bone resorption, i.e. osteoclast surface, was considerably increased and osteoid volume moderately increased. The remaining histomorphometric parameters studied were generally normal for age and sex as compared to the published reference data. Increased osteoclast surface in 65% of patients indicated that bone loss was an active and prevailing process in these postmenopausal women, which was considerably more pronounced than in the normal age-matched population. Results of the histomorphometric analysis were categorized according to the published classification of postmenopausal osteoporosis. The percentage of patients in each group differed from literature data, most probably due to the sample size and choice. None of the patients had histomorphometric features of reduced osteoblastic and osteoclastic activity, but in 37% of postmenopausal women osteoclastic activity was increased while osteoblastic activity was normal, a feature not described in the original histomorphometric classification of postmenopausal osteoporosis. CONCLUSIONS: Histomorphometric analysis of bone biopsy in women with postmenopausal osteoporosis revealed bone resorption as a predominant finding. Different groups were recognized based on the diversity of bone cell activity. The difference in the frequencies in study groups, and observation of a distinct group not included in the histomorphometric classification of postmenopausal osteoporosis probably resulted from sample size and nonspecific population traits. Histomorphometric analysis of bone in postmenopausal osteoporosis is an important contribution to better understanding of this most common bone disorder. PMID- 15125388 TI - [Intermittent intravenous administration of cyclophosphamide in patients with lupus nephritis]. AB - PATIENTS AND METHODS: Patients with systemic lupus erythematosus (SLE) and lupus nephritis submitted to renal biopsy were retrospectively analyzed. All patients were admitted to Dubrava University Hospital, Department of Clinical Immunology and Rheumatology and Department of Nephrology. The diagnosis of SLE based on the criteria proposed by the American Rheumatology Association was made in 46 patients treated during the last 4 years. There were 39 female and seven male patients, mean age 39.5 years. Renal biopsy was performed in 16 patients (three male and 13 female, mean age 34 years). RESULTS: Diffuse proliferative glomerulonephritis (type IV) was diagnosed in ten, focal segmental glomerulonephritis (type III) in three, and mesangioproliferative glomerulonephritis (type II) in two patients. The type of lupus nephritis was classified according to the World Health Organisation (WHO) criteria. In one patient, the tissue obtained proved inadequate for histologic analysis. Pulsed cyclophosphamide therapy was indicated in patients with lupus nephritis type III and IV (WHO). Cyclophosphamide was administered at a dose of 500-1000 mg i.v.. All patients received corticosteroids. One patient was administered intravenous immunoglobulin 400 mg/kg body weight. In ten out 13 patients with lupus nephritis types III and IV, the therapeutic protocol was completed, while the treatment is still under way in the remaining three patients. In nine and four of 13 patients, complete and incomplete remission was achieved after 6-month treatment, respectively. Two of 15 patients with type II mesangioproliferative glomerulonephritis were treated only with corticosteroids alone. CONCLUSION: We believe that intermittent pulsed cyclophosphamide therapy produces favorable effects in the management of lupus nephritis type III and IV. PMID- 15125389 TI - [Value of urinary sediment cytology in evaluation of acute tubular necrosis after kidney transplantation]. AB - AIM: The aim of the study was to assess the clinical value of urinary sediment cytology (USC) by use of phase-contrast microscopy in the evaluation of acute tubular necrosis (ATN) during the early period after kidney transplantation. The study was performed at the Cytology Laboratory, Department of Nephrology and Dialysis, Clinical Hospital Center Rijeka, Croatia. PATIENTS AND METHODS: Patients included 141 kidney recipients, 99 male and 42 female, mean age 40 +/- 13 (range 8-72) years, who had received kidney allograft during the period of ten years, and who were treated at the University Department of Internal Medicine, Rijeka Clinical Hospital Center. The majority of patients (76%) had received cadaveric kidneys. Urinary sediment was analyzed for the presence of renal tubular cells, isomorphic erythrocytes, lymphocytes, casts and debris. Renal tubular cells on USC were recognized as the most constant sign of ATN. The presence of lymphocytes should arise suspicion of rejection. MAIN OUTCOME MEASURES: A typical cytologic profile of acute tubular lesion consists of tubular cells, isomorphic erythrocytes, casts, cellular and/or amorphic debris. RESULTS: USC by use of phase contrast microscopy is confirmed as a method of a very high sensitivity (82%) and specificity (93%) in the evaluation of ATN in transplanted kidney patients during early post-transplantation period. In situations of coexistence of several causes of allograft dysfunction, "mixed" cytologic pictures were frequently created, from which it is difficult or almost impossible to identify the actual cause of kidney dysfunction. In these cases, the final judgment should be made solely by histologic evaluation, which still represents the gold standard in the evaluation of kidney allograft dysfunction. CONCLUSION: Serial USC, when thoroughly examined using phase-contrast microscopy, is a simple, noninvasive, fast, easily repeatable and inexpensive diagnostic method of high sensitivity and specificity in the evaluation of ATN during the early phase after kidney transplantation. PMID- 15125390 TI - [How much body water volume affects evaluation of urea clearance in patients treated with peritoneal dialysis?]. AB - AIM OF THE STUDY: Peritoneal dialysis adequacy is usually estimated using the ratio of total weekly urea clearance (Kt) and urea distribution volume (V), the later being identical to total body water volume. It is observed that even patients with acceptable Kt/V values sometimes show signs and symptoms of dialysis inadequacy. A question arose whether the discrepancy came from a falsely assessed urea distribution volume, because the less the urea distribution volume measured the higher the Kt/V gained. PATIENTS: The survey included 32 patients (15 women, 17 men) in whom 62 total weekly urea clearances were calculated during a 2-year period. Each Kt was used in 4 different equations (A-Watson's method, B Hume's method, C-58% of body mass for urea distribution volume, D-for urea distribution volume: 58% of body mass reduced by 2 kg, and then augmented by 2 kg added for dialysate volume assumed). It resulted in 4 different Kt/V for each Kt, i.e. a total of 248 Kt/V values. RESULTS AND DISCUSSION: At the moment of the examination the patients were aged 57.66 +/- 8.66 years, weighted 75.54 +/- 11.04 kg (with dialysate intraperitoneally), and were treated with peritoneal dialysis for a mean of 23.28 +/- 27.55 months. Their mean total weekly urea clearance was 79.94 +/- 12.53 L. Depending on the method used, the mean urea distribution volumes were as follows: A = 37.24 +/- 5.27 L, B = 37.89 +/- 5.22 L, C = 43.81 +/ 6.40 L and D = 44.65 +/- 6.47 L, and mean Kt/V 2.15 +/- 0.32 (A), 2.11 +/- 0.32 (B), 1.82 +/- 0.33 (C) and 1.79 +/- 0.34 (D). The highest mean urea distribution volume (D) was higher than the lowest value (A) by 19.90%, and the difference between those values was statistically significant (tV D: A = 6.99, p < 0.01). The highest mean total weekly Kt/V (A) was higher than the lowest value (D) by 20.11%, and the difference was also significant ((tKt/V A: D = 6.08, p < 0.01). CONCLUSION: The estimation of urea distribution volume has a direct impact on calculation results of total weekly Kt/V in peritoneal dialysis patients. Urea distribution volumes as assessed by the anthropometrical methods or by a certain percentage of body mass were approximate values. We suggest replacing "nondeuterial methods" by deuterial, in order to get more comparable results. PMID- 15125391 TI - [Comparison of biocompatibility of hemophane, cellulose diacetate and acrilonitile membranes in hemodialysis]. AB - INTRODUCTION: The membranes used in haemodialysis (HD) may be manufactured from cellulose, modified cellulose or synthetic polymers. Such membranes, when in contact with blood, activate the complement system, which entails changes in leukocyte (L) and platelet (P) counts. The magnitude of complement activation depends on the type of membrane used, and represents the biocompatibility index. Acrylonitrile (AN69) is synthetic membrane of high biocompatibility standards, whereas haemophane (HP) and cellulosediacetate (CD) membranes are modified cellulose-based membranes. The biocompatibly profiles of HP, CD and AN69, characterised by changes in L and P counts and activation of complement components (C3a and C5b-9), has been studied in order to assess the biocompatibility features of HP, CD and AN69 membranes. MATERIALS AND METHODS: We compared the generation of C3a des Arg and C5b-9 and the changes in L and P counts in groups of patients on HD who used HP (3 patients), CD (3 patients) and AN69 membrane (3 patients). Concentrations of C3a des Arg and C5b-9, and L and P counts were measured in the first HD session of the week, just before the start of HD (0 min) and after 15, 120 and 240 min. RESULTS: For HP, L count at 15 min had declined to 50% of the pretreatment level (p < 0.05), compared with 59.3% (p < 0.05) for CD and with 98.7% (p = ns) for AN69, returning to the pretreatment level at 240 min. Statistically significant difference was found comparing L counts for HP vs AN69 (p < 0.001) and CD vs AN69 (p < 0.001). The mean plasma concentrations of C3a des Arg were 1246 +/- 832 ng/ml for HP, 1148 +/- 774 ng/ml for CD and 639 +/- 217 ng/ml for AN69 and significant difference was found comparing HP vs AN69, CD vs AN69 (p < 0.05). The maximal values of C3a des Arg occurred at 15 min in HP and CD (p < 0.005), whilst for AN69 the concentrations showed no statistically significant differences. The plasma concentrations of C5b 9 in patients on HP and CD were the highest at 15 min (p < 0.005), while those on AN69 did not show any statistically significant rise. DISCUSSION: The fall in L counts was considerably marked for HP and CD membranes, with the lowest level at 15 min following blood exposure, thereafter returning to pretreatment level, whilst in AN69 leukopenia was not noted. The loss of L from the circulation is attributed to their pulmonary sequestration since complement activation and release of C5a fraction into the circulation results in their binding to the neutrophils and upregulation of the adhesion-promoting integrins of leukocytes, resulting in their binding to endothelial cells. The mean plasma concentrations of C3ades Arg, measured in the course of clinical use of HP and CD were significantly higher compared with synthetic membrane AN69, reaching the maximal level at 15 min. The complement activating potential of a biomaterial is one of its important biocompatibility features. The C3a complement activation curve is characteristic of cellulose-based materials and is governed by interaction between blood and hydroxyl groups on the membrane surface and by the density of hydroxyl groups available for the binding of C3b. The highest concentrations of C5b-9 in modified cellulose membranes were achieved by 15 min following blood exposure and met the activation curve for C3a. The capacity of AN69 membrane to adsorb anaphylatoxins may be a contributory factor for the failure to measure any increase in complement fraction concentration during HD for AN69. CONCLUSION: There are significant differences between HP, CD and AN69 membranes concerning complement activation magnitude and leukopenia rate. Modified cellulose membranes HP and CD induce higher C3a concentrations and superior leukopenia rate compared with AN69. Synthetic membrane AN69 proved of higher biocompatibility, with lower efficiency of complement activation. PMID- 15125392 TI - [Knowledge and attitudes on breastfeeding among parturients]. AB - During human history, the natural process of breastfeeding has become ever less present as a mode of infant feeding. Numerous incentives have been introduced worldwide to stop this unfavorable trend. AIM OF THE STUDY: The aim of the study was to find the basic attitudes and knowledge about breastfeeding among mothers after delivery while they were at maternity wards. METHODS: Two surveys were conducted, with 940 participants in 1989-1990, and 626 participants in 1997. The women answered a self administered questionnaire with multiple choice questions. Data from the 1989-1990 and 1997 surveys were analyzed and compared. RESULTS AND DISCUSSION: A high percentage of women (98%), stated that they wanted to breastfeed their children, but only about 44% of them expected it to be problem free. A great number of women did not know how and when to estimate milk secretion, or its adequacy for the baby. A significant proportion of the mothers believe that milk of some women is "watery" and thus inappropriate for their children. A great number of mothers (83%), answered correctly that the child need to be fed on demand rather than according to a strict schedule. In the second phase of the survey some positive changes in their knowledge were found, however, yet quite inadequate. CONCLUSION: Comprehensive and continuing education about breastfeeding of future mothers and fathers and the society as a whole is required. PMID- 15125393 TI - [Ultrasonography methods in the diagnosis of renal osteodystrophy]. AB - Bone disease, i.e. renal osteodystrophy, is commonly seen in patients with chronic renal failure. It encompasses all the disorders of mineral and bone metabolism associated with chronic renal insufficiency, i.e. secondary hyperparathyroidism, retention and accumulation of beta 2 microglobulin and aluminum. The most frequent cause of renal osteodystrophy is secondary hyperthyroidism, with a consequence of high turnover bone disease. Secondary hyperparathyroidism, i.e. increased parathyroid hormone (PTH) secretion and parathyroid gland hyperplasia, develops early in the course of chronic renal insufficiency. Hypocalcemia, phosphate retention and deficiency of calcitriol stimulate PTH synthesis and secretion and parathyroid cell proliferation, i.e. hyperplasia. Parathyroid cell proliferation is initially polyclonal (diffuse hyperplasia), and later it is monoclonal or multiclonal (nodular hyperplasia). Calcitriol receptors as well as calcium-sensing receptors are significantly reduced in parathyroid glands in nodular hyperplasia. Patients with such parathyroid gland hyperplasia are often resistant to vitamin D therapy. A specific form of bone disease is beta 2 amyloidosis. Destructive arthropathy, cystic changes and carpal tunnel syndrome are clinical manifestations of dialysis related amyloidosis, which is one of the major complications in patients on longterm hemodialysis. Aluminum intoxication leads to the low turnover bone disease and consequential osteomalacia or aplastic bone lesions, the cause of which has not yet been fully clarified. Ultrasound can be a useful, economical and noninvasive method in the evaluation of renal osteodystrophy. Ultrasound waves are very important for noninvasive imaging of soft tissue, especially parathyroid glands, pathologic changes of the joints, and for detection of metastatic calcifications. They are also useful in the evaluation of skeletal status in dialysis patients. Ultrasound waves of a frequency above the limit of human hearing are used in the morphological diagnosis of parathyroid gland. Today, because of its simplicity and non-invasiveness, it is a generally accepted method for the detection of enlarged parathyroid gland in patients with secondary hyperparathyroidism, for the monitoring of pathologic changes, and for making decisions on the method of treatment based on the size and number of parathyroid glands. Ultrasound can distinguish nodal from diffuse parathyroid hyperplasia. Under ultrasound guidance it is possible to perform fine needle aspiration biopsy, to confirm ultrasound findings, and percutaneous inactivation of parathyroid gland (PEI) with alcohol. Ultrasound is useful in the diagnosis of pathologic changes of the musculoskeletal system in patients with beta 2 amyloidosis, to assess the process of its spread, especially in the shoulder joint where the changes are most pronounced (rotator cuff thickness, amyloid deposits as hyperechogenic pads, and detection of fluid in the joint), but it can also be used to examine other joints as well as soft tissue in which metastatic calcifications may occur. Standard ultrasound equipment (pulse-echo) and linear probe of 5-13 MHz are used, also serving for ultrasound examination of the neck, joints and soft tissue. Quantitative bone ultrasonometry is based on different physical characteristics of the ultrasound including: transmission, Speed Of Sound (SOS) in meters/sec and Broad Band Attenuation (BUA) in dB/MHz, and different concepts of the apparatus. These parameters depend on the strength and architecture of the bones and describe better the changes in bone structure in dialysis patients by calculation of the Stiffness Index (QUI), better than the standard bone densitometry by dual-energy x-ray absorptiometry, which only measures bone density. Combined ultrasound measurement of the bone in several locations may be successful in monitoring dialysis patients. PMID- 15125394 TI - [Importance of biochemical indicators of bone turnover in patients on chronic dialysis and after kidney transplantation]. AB - Bone remodeling is a continuous process of removal of microscopic amounts of bone tissue due to synchronized actions of osteoclasts and osteoblasts with the purpose of renewal and repair of bone tissue. During the formation of bone matrix osteoblasts synthesize proteins. Measurement some of these proteins in blood has clinical significance as indicators of bone formation: osteocalcin, procollagen type I propeptide, and bone alkaline phosphatase. During osteoclastic bone resorption, collagen type I breakdown fragments are released in the circulation and excreted in the urine, and measured in serum or urine as bone resorption markers (telopeptide, pyridynolines). Bone metabolism and accordingly bone markers are subjected to considerable biologic variation. The effects of age, sex, race, pregnancy and lactation, fracture, disease and certain drugs cannot be avoided and must thus be considered when interpreting the results. Circadian variation is excluded by obtaining samples in the morning and the effect of exercise prevented. The use of bone markers has been extensively studied in monitoring the effect of antiresorptive treatment in osteoporotic women. A decrease of 30-50% occurs within 3 months after the beginning of hormone replacement therapy or bisphosphonates and remains at this level. In patients on chronic dialysis treatment, bone markers are increased and reflect bone metabolism as assessed by bone biopsy. Although bone markers enable discrimination between high and low bone turnover, they cannot substitute for bone biopsy in determination of the type of renal osteodystrophy. The factors affecting bone disorder in these patients, i.e. dialysis duration or parathyroid function correlate with bone markers. In kidney transplant recipients, an increased bone turnover and its normalization after approximately 2 years can be assessed by bone markers. Similar to chronic dialysis, risk factors for bone disorder after kidney transplantation (e.g., dialysis duration, parathyroid function, age, sex, immunosuppressants, corticosteroids, graft function) are associated with bone markers. We present cross-sectional and longitudinal data on 100 patients on chronic dialysis and 80 kidney transplant recipients. In conclusion, sufficient evidence exists indicating that the measurement of bone markers enables assessment of bone turnover and its dynamics. However, no guidelines or recommendations have been put forward to validate their use in routine clinical practice of chronic dialysis or kidney transplantation bone disorders. PMID- 15125395 TI - [Uremic polyneuropathy]. AB - Uremic polyneuropathy is probably the most common complication of chronic renal failure. About 70 percent of dialysis patients regularly have uremic polyneuropathy, in 30 percent the neuropathy is moderate or severe. Coexistence of muscle weakness and atrophy, areflexia, sensory loss and graded distribution of neurologic deficit in a patient with renal disease suggests the presence of uremic polyneuropathy. During longterm hemodialysis, the symptoms of polyneuropathy stabilize, but they improve only in relatively few patients. Complete recovery, occurring over a period of 6 to 12 months, usually follows successful renal transplantation. PMID- 15125396 TI - [Importance of plasmapheresis in the treatment of paraneoplastic cerebellar degeneration]. AB - Paraneoplastic neurologic syndromes are disorders of the nervous system function caused by cancer but not due to metastatic disease, vascular or metabolic deficits, infections, nutritive deficiency, nor side effects of antineoplastic drugs or irradiation. Immunologic factors probably play the crucial role in the pathogenesis of paraneoplastic neurologic syndromes, but nonimmunologic mechanisms that include metabolic abnormalities and competition for substrate are also involved. Paraneoplastic cerebellar degeneration most commonly occurs in the setting of gynecologic cancers, but it accompanies the small-cell lung cancer too. Other tumors are infrequently associated with cerebellar degeneration. Several paraneoplastic antibodies have been identified in patients with paraneoplastic cerebellar degeneration. Their association with particular cancers may help identify an occult lesion. Anti-Yo antibodies are directed against Purkinje cell antigens and occur in patients with cerebellar degeneration who have breast cancer or gynecologic tumors. A target antigen of anti-Yo antibody is CDR2 protein that is normally expressed only in the brain and testis. Patients with paraneoplastic cerebellar degeneration present with dizziness, nausea and vomiting followed by gait instability, diplopia, gait and appendicular ataxia, dysarthria and dysphagia. Therapeutic options include tumor excision, chemotherapy and/or irradiation, and adjuvant therapy with glucocorticoids, immunoglobulins and plasmapheresis. The role of plasmapheresis in the treatment of paraneoplastic cerebellar degeneration is still uncertain. Reports of its efficacy are anecdotal. We present patient with paraneoplastic cerebellar degeneration with positive anti-Yo antibodies and tumor of the ovaries whose neurologic status significantly improved after four daily plasmaphereses, which was accompanied by a fourfold decrease in the anti-Yo antibodies titer. Further investigations are needed to define a protocol for plasmapheresis in the treatment of patients with paraneoplastic syndromes. PMID- 15125397 TI - [Work capacity in patients on hemodialysis]. AB - The quality of life is considerably impaired in patients on regular hemodialysis has been changed. It is difficult to determine it because there are no general definitions or measuring instruments. There are objective and subjective components of the quality of life, one among them being occupational ability. During the progression of chronic renal disease (CRD) to terminal renal failure (TRF) physical activity of the patients becomes poorer. In this stage, their physical activity is by 40-60% below the value expected for the same healthy age cohort. The intention of this analysis was to determine occupational ability in patients on regular hemodialysis. The analysis included 161 patients on hemodialysis, 78 (48.5%) female and 83 (51.5%) male, mean age 61.2 +/- 13.1 years, and mean time on hemodialysis was 54 +/- 71.9 months. All patients filled out a self-administered questionnaire on schooling and occupational ability. The cause of TRF was glomerulonephritis in 45 (26.8%), diabetes mellitus in 42 (26.3%), nephrosclerosis in 26 (16.1%), and pyelonephritis in 12 (7.4%) patients. Age distribution was as follows: 0-19 years 1 patient, 20-44 years 14 (8.7%); 45 64 years 64 (39.8%) and 65 years 82 (50.9%) patients. Educational structure: elementary school 65 (40.4%), secondary school 79 (49.1%), college 10 (6.2%), and university 6 (3.7%) patients. Occupational structure: retired 123 (76.4%), housekeeper 20 (12.4%), never employed 4 (2.5%), employed 10 (6.2%), unemployed 2 (1.2%), 1 child and 1 student. Among employed patients there were 7 men and 3 women. Their educational level was as follow: elementary school 1 patient, secondary school 8 patients, college 1 patient. At the beginning of hemodialysis their occupational status was: full-time employment 30 (18.6%) patients, part time employment 1 patient, longer time on sick-leave payment (3.1%), retired 95 (59%), pupils and students 3, unemployed 2, and 1 child did not attended school. Time interval between the beginning of hemodialysis and retirement was: less than 1 year work 13 (36.1%) patients, 1-2 year work 6 (16.7%), three year work 2 patients, more than 8 year work 2 patients, and 10 year work only 3 patients, for 14, 18 and 26 years each. Two patients lost their job for employer bankruptcy. The judgment of patients regarding their occupational ability was as follows: out of 161 patients, 23 (14.3%) felt fit for work, 12 on full-time and 11 on part time basis. Occupationally incapable were 46.6% of patients, and 63 felt unable to take care of another person. Some kind of additional activity, like working in garden or taking care of children was reported by 26 patients. The aforementioned results showed that 22.4% of the patients were occupationally active at the time of starting hemodialysis. Many patients were retired after hemodialysis had started. Only 6.2% of hemodialysis patients were occupationally active although 14.3% felt occupationally capable. The main reasons for such a low level of employment were advanced age, diminished physical activity due to the disease, and difficulties associated with the socioeconomic situation in the country. PMID- 15125399 TI - Percutaneous interventions in stenotic valvular heart disease complicating pregnancy. PMID- 15125398 TI - [Incidence of chronic renal failure during 35 years at the Rijeka Clinical Hospital Center]. AB - Chronic hemodialysis treatment in the world started in 1960. At that time, due to technical development and construction of arteriovenous shunt for repeated blood access for dialysis, it was possible to treat uremia. At the Department of Surgery, Rijeka Clinical Hospital, former Dr. Zdravko Kucic Hospital, first hemodialysis was performed in 1962, whereas regular chronic dialysis treatment started in 1966. On September 20, 1966, the first hemodialysis was done in a patient with chronic uremia. A week later, the next patient was admitted for therapy. The aim of the study was to analyze demographic and other data of all patients who started dialysis in the period between 1966 and 2001. There were 910 patients, 555 (60.9%) male and 355 (39.1%) female. In the first year, only two patients were treated with hemodialysis. Until 1970, the number of new patients was 4 or 5. From 1971 till 1984 between 10 and 19 new patients started dialysis every year, and from 1985 till 1990 their number ranged from 23 to 34 per year. Further increase in the number of treated patients was recorded in 1990 when 42 patients were dialyzed. In the following years until 2001 the figures were between 40 and 58, and in 2001 55 new patients were treated. In 1966, the mean age of patients undergoing this therapy was 29.5 years, and with time it increased to 40 in 1978. From 1989 on, the mean age rose to 50, and from 1998 to 60. In 2001, the mean patient age was 63.2 years. The primary renal diseases that led to uremia in the study population were glomerulonephritis (GN) in 256 (28.1%), pyelonephritis (PN) in 165 (18.1%), diabetes mellitus (DM) in 161 (17.7%), nephrosclerosis in 111 (12.2%), uremia after transplanted kidney rejection in 47 (5.2%), polycystosis in 40 (4.4%), lupus nephritis in 12 (1.3%), other causes in 89 (9.7%), and unknown cause in 24 (2.6%) patients. The distribution of primary renal disease during the observed period was as follows: from 1966 till 1979 the cause of uremia was GN in 88 (62%), PN in 30 (21.1%), DM in only 1, polycystosis in 3, post-transplant uremia in 7, lupus in 3, and other causes in 7 patients. From 1980 to 1989, GN was the cause of uremia in 67 (31.6%), PN in 45 (21.2%), DM in 22 (10.4%), nephrosclerosis in 26 (12.3%), polycystosis in 11 (5.2%), post-transplantation uremia in 12 (5.7%), lupus nephritis in 8 (3.8%), other causes in 17 (7.9%) and unknown cause in 3 (1.9%) patients. During the 1990-2001 period, GN was recorded in only 101 (18%), PN in 90 (16.2%), DM in 138 (24.9%), nephrosclerosis in 82 (14.7%), polycystosis in 26 (4.7%), post-transplantation uremia in 28 (5.0%), lupus nephritis in 6 (1.1%), other causes in 65 (11.7%) and unknown cause in 20 (3.6%) patients. The mortality was caused by cardiac disease in 50.4%, cerebrovascular disease in 14.8%, infectious disease in 13.2%, malignancy in 7.5%, high potassium in 5.1%, gastrointestinal disease in 3.5%, other vascular diseases in 1.6%, cachexia in 1.3%, loss of blood access in 0.8%, other reasons in 1.1% and unknown reasons in 0.5% of patients. The results clearly indicate that the number of new patients grew and the mean patient age increased every year. Diabetes mellitus was the leading cause of uremia while GN and PN were less common. The main causes of death were cardiovascular diseases. PMID- 15125400 TI - Spiritual health. PMID- 15125401 TI - Nutritional status and productivity of Sri Lankan tea pluckers. AB - OBJECTIVE: To assess the impact of nutritional status on productivity of tea pluckers. DESIGN: Cross-sectional, analytical. SETTING AND PARTICIPANTS OF STUDY: All tea pluckers in five divisions of a tea estate in Hatton. MEASUREMENTS: A dietary survey was conducted using the 24 h dietary recall method. Nutritional status was assessed by measurement of body mass index (BMI) and haemoglobin (Hb) levels, and faeces were examined for helminth ova. The daily weight of tea leaves plucked and the number of days worked by each woman during the preceding month were noted from production records. RESULTS: Three hundred and four women (mean age 37.8 years, SD 8.4) were examined. The majority (59.9%) had evidence of chronic energy deficiency (BMI < 18.5). Almost all (94.4%) were anaemic (Hb < 13.3 g/dl, altitude adjusted cutoff). Mean daily iron intake was 8.7 mg (SD 2.3 mg), 45.9% of the national recommended dietary allowance. Only 10.1% had hookworm infection, all of light intensity. Multivariate regression analysis showed that dietary iron intake, the number of children and the number of children below 5 years, but not hookworm infection, were independently associated with Hb (p < 0.05). There was a strong positive correlation between monthly productivity and Hb, but not with BMI. Variation in Hb levels accounted for 65% of variation in productivity (adjusted R2 = 0.651). CONCLUSIONS: Most of the study population had chronic energy deficiency and anaemia. Dietary inadequacy of iron is a much more important causative factor than hookworm infection. Productivity was strongly associated with the degree of anaemia but not with BMI. PMID- 15125402 TI - Varicella-zoster virus infection in the Infectious Diseases Hospital, Sri Lanka. AB - OBJECTIVES: To determine the morbidity and mortality patterns of varicella and risk factors affecting its outcome, and the facilities available at the Infectious Diseases Hospital (IDH), Sri Lanka. METHODS: A retrospective study on all patients admitted with varicella-zoster virus (VZV) infection to the IDH from August 2000 to July 2001. Data were collected from the hospital records. RESULTS: Among the 1690 patients admitted during the study period, 1090 (64.9%) were due to VZV infection. Nine hundred and eighty nine (90.7%) had varicella and 101 (9.3%) herpes zoster. Common complications were secondary bacterial infection (62.1%), neurological complications (3.4%), pneumonia (9.1%) and carditis (1.01%). They were significantly commoner in patients with coexisting diseases. Hospital stay was significantly shorter in patients who received early aciclovir, which was not available on a regular basis. Forty one patients died and mortality was highest in the elderly. The commonest cause of death was pneumonia. CONCLUSIONS: Varicella related complications are high in patients with coexisting diseases. Mortality rates are higher than reported elsewhere. Health care facilities available at IDH are quite inadequate, and should be improved. PMID- 15125403 TI - Detection and management of alcohol misuse by general practitioners. AB - INTRODUCTION: Alcohol misuse and related problems are common in Sri Lanka. The appropriateness of the primary care setting in dealing with alcohol misuse is well recognised, and general practitioners (GPs) constitute an important first contact setting. METHODS: One hundred and fifty randomly selected GPs practising in the Colombo and Gampaha districts were given a questionnaire to assess how they detect and manage alcohol misuse, and their attitudes towards persons who misuse alcohol. RESULTS: Seventy per cent of GPs responded [74 male; mean age 42 years (SD 6.7)]. Our results suggest that although a majority (81%) of GPs were frequently confronted with problems related to alcohol misuse, their efforts to detect the problem and knowledge regarding risk limits of alcohol consumption were poor. Only 25.7% had even heard of CAGE and MAST questionnaires. The majority of GPs felt inadequately trained to deal with alcohol misuse, but only a few made any self-directed efforts to improve their knowledge and skills regarding its management or referred their patients for specialised care. Participation in preventive programmes was minimal, and many GPs expressed negative attitudes towards persons misusing alcohol. CONCLUSIONS: The ability of general practitioners to detect and alcohol misuse appears to be inadequate. PMID- 15125404 TI - Changing patterns of thyroid cancer in Sri Lanka. Has the iodination programme helped? AB - OBJECTIVES: To ascertain if there has been a change in the pattern of thyroid cancer in Sri Lanka. If so, whether there is a correlation to the implementation of the programme of iodination. DESIGN: Retrospective (1974-1986) and prospective (1987-2001). SETTING: Kandy Hospital (1974-1982), Peradeniya Hospital (1982-2001) and private hospitals in Kandy (1979-2001). METHOD: Three hundred consecutive patients with cancer of the thyroid seen over 28 years (1974-2001) period were reviewed for demography, histopathology and extent of spread at presentation. Seventy one patients of this group had a pre-existing goitre of greater than 10 years' duration and were similarly reviewed. RESULTS: A highly significant reduction of anaplastic thyroid cancer and a highly significant reduction in the extent of extra-thyroidal spread at presentation of differentiated thyroid cancer were observed after 1995. In malignancy supervening on pre-existing goitre, a significant reduction in anaplastic carcinoma and a highly significant increase in papillary carcinoma were noted in the post-1996 period. A significant reduction of extra-thyroidal spread was also observed. CONCLUSION: A trend towards more differentiated thyroid cancer with lesser degree of spread was observed in recent years. The iodination programme implemented in 1995 is likely to be responsible for this change. PMID- 15125405 TI - Prenatal testing and pregnancy termination in Sri Lanka: views of medical students and doctors. AB - INTRODUCTION: Prenatal diagnosis and pregnancy termination generate complex ethical issues. Surveys conducted in Sri Lanka among doctors and medical students in 1986 supported a change in law in favour of pregnancy termination when gross genetic defects are detected antenatally. A new generation of prenatal tests has focused attention again on the topic of termination and under what circumstances it might be legally done. The present survey contributes to the debate by means of a survey of doctors and medical students. METHOD: A self-administered questionnaire given to doctors and medical students. RESULTS: Ninety three per cent of doctors and 81% of students accept pregnancy termination as an appropriate course of action if a gross genetic defect is detected antenatally, and 87% of doctors and 80% of students support a change in the law to allow termination of the pregnancy. The corresponding figures in previous surveys were 80%, 69% and 96%, 88% respectively. CONCLUSIONS: A majority of doctors and medical students support a change in law in favour of liberalising pregnancy termination when a genetic defect is detected antenatally. PMID- 15125406 TI - Parkinson disease of ranking lawyer and legislator SJV Chelvanayakam: a hypothesis. AB - SJV Chelvanayakam (1898-1977), a ranking civil lawyer and legislator, was probably the well known Parkinson disease victim in the 20th century Sri Lanka. He was born in Ipoh, Malaya, where his father had moved in the last decade of the 19th century for professional advancement. Ipoh was then an attractive location for migrants from China and the Indian subcontinent since it was in the Kinta valley--touted then, as the world's richest single tin field. Chelvanayakam was brought to Jaffna peninsula when he was aged four (in 1902 or 1903) by his mother, who returned to her native Tellipalai town partly due to indifferent health during her stay in Kinta region. In this communication, I present a hypothesis that organotin exposure as a foetus or during infancy at his place of birth is likely to have been a contributing factor to Chelvanayakam's Parkinsonism. It seems to fit the available circumstantial evidence. PMID- 15125407 TI - Qualitative research. PMID- 15125408 TI - Young patients with movement disorders. PMID- 15125410 TI - Dengue encephalitis in a child. PMID- 15125409 TI - A case of primary neuroleptospirosis. PMID- 15125411 TI - Severe multi-organ dysfunction following multiple wasp (Vespa affinis) stings. PMID- 15125412 TI - A case of self-limiting Coomb's negative haemolytic anaemia following dengue shock syndrome. PMID- 15125413 TI - Safety and efficacy of subcutaneous adrenaline as a treatment for anaphylactic reactions to polyvalent antivenom. PMID- 15125414 TI - Health professionals and torture. PMID- 15125415 TI - Legal and ethical considerations of "living will". PMID- 15125416 TI - [Theriac: medicine and antidote]. AB - Theriac was an ancient multi-ingredient preparation; originating as a cure for the bites of serpents, mad dogs and wild beasts, it later became an antidote to all known poisons. The name theriac (treacle), (Greek theriake, Latin theriaca, French theriaque) was derived from the Greek for wild beast - theriakos. The first formula was created by Mithridates Vl, King of Pontus, a skillful ruler but a monster of cruelty, who, living in such a fear of being poisoned, took a great interest in toxicology. In the 1st century AD, Nero's personal physician Andromachus improved the formula of Antidotum Mithridatium by adding flesh of vipers, which was commonly believed to be the best antidote against snakebite, and by increasing the proportion of opium. It became known as Theriac of Andromachus, and contained 64 ingredients including various minerals, herbals, poisons and animal flesh and blood, all combined with honey in the form of electuarium. Later it became the cure-all medicine which, accumulating all the simples into one form, was supposed to be a universal panacea against all diseases. In the Middle Ages this famous electuarium become a patent medicine and entered official dispensaries and pharmacopoeias. The most famous and expensive Theriac in Europe was that of Venice. It was not until the l8th century that it was excluded from medical use. PMID- 15125417 TI - The UNESCO concept of heritage and teaching of the history of medicine and pharmacy in the Karol Marcinkowski University of Medical Sciences in Poznan. AB - This article points to the possibility of the creative use of the UNESCO concept of heritage in the teaching of the history of medicine and pharmacy. This enriching teaching concept is of great importance, with 2002 being proclaimed the UNESCO Year for Cultural Heritage. The article discusses long-term experiences gathered at the Karol Marcinkowski University of Medical Sciences in Poland. PMID- 15125418 TI - The state of medical history in Australia. PMID- 15125419 TI - A model from Turkey on the history of the health sciences as seen from the point of view of scientific societies and their contributions-the Guest House Society. PMID- 15125420 TI - Labor-saving technology and fertility increase in rural Africa. PMID- 15125421 TI - Benchmark your financial fitness. PMID- 15125422 TI - UAP delegation: a step-by-step process. PMID- 15125423 TI - [Possibilities of optimization of the system of medical rehabilitation of military servicemen after coronary artery bypass surgery]. PMID- 15125424 TI - [Long-term results of surgical treatment of the cardiac postinfarction aneurysms by restoration of the volume and geometry of the left ventricle]. PMID- 15125425 TI - [Nifedipine GITS--clinical efficacy and effect on quality of life in elderly patients with hypertension]. PMID- 15125427 TI - Abstracts of the 21st Congress on Clinical Hemorheology and Microcirculation of the German Society. Ulm, Germany, 20-25 October 2002. PMID- 15125426 TI - HFMA: hospital capital spending lags behind depreciation. PMID- 15125428 TI - [Effect of high doses of broncholytics on the state of cardiovascular system during treatment of sever exacerbation of bronchial asthma]. AB - The use of broncholytic drugs through nebulizer is a standard treatment for acute episodes of bronchial asthma. However doses of these drugs administered by nebulizer can be much higher than those applied when metered dose inhaler is used. These doses have a potential of cardiotoxic effects. MATERIAL: Patients (n=75) with severe exacerbation of bronchial asthma. METHODS: All patients received broncholytic drugs through nebulizer. Doses were adjusted according to age and concomitant pathology among which prevailed ischemic heart disease (21.3%) and hypertension (53.3%). Nebulized beclomethasone was an alternative to systemic corticosteroids. Safety of therapy was controlled with electrocardiography, 24-hour electrocardiography, measurements of serum potassium level, registration of subjective signs (palpitation, tremor) and objective data (heart rate, blood pressure). RESULTS: Clinical improvement occurred in all patients. By the end of nebulizer therapy arterial blood O(2) saturation increased 4.66% from initial level (p=0.04). Peak expiratory rate and forced expiratory volume in 1 sec also rose (p <0.05). Termination of nebulizer therapy was associated with decrease of number of both supraventricular and ventricular premature beats. Moderate widening of of QT interval above 460 ms was registered in 4 patients, 3 of whom had ischemic heart disease. There were no significant changes of serum potassium levels. CONCLUSION: Broncholytic drugs administered by nebulizer in therapeutic doses selected with consideration of age and concomitant diseases did not produce cardiotoxic effects. PMID- 15125429 TI - [Characteristics of clinical and pharmacological efficacy of tranquilizers and antidepressants in patients with ischemic heart disease and type A behavior]. PMID- 15125434 TI - [Fibrates: mechanism of action, effect on levels of lipids and lipoproteins, risk of coronary events. Part 1. Clofibrate, gemfibrosil, besafibrate]. PMID- 15125435 TI - [Pharmaco-economical aspects of reduction of left ventricular hypertrophy in hypertensive patients: how we can use the results of clinical trials]. PMID- 15125436 TI - [Pharmaco-economical aspects of the treatment of patients with chronic cardiac failure in hospital]. PMID- 15125437 TI - [Extraordinary case of hemodynamic correction of dilated cardiomyopathy]. PMID- 15125438 TI - Bibliography. Current world literature. Crystal deposition diseases. PMID- 15125439 TI - Abstracts of the 2004 Meeting of the Society for Technology in Anesthesia, the 2004 International Meeting on Medical Simulation, and XXIV Computers in Anesthesia Meeting. PMID- 15125440 TI - Patent news. PMID- 15125441 TI - Patent alert. PMID- 15125442 TI - [DEGRO 2003. Abstracts of the 9th Annual Congress of the German Society of Radio Oncology. Essen, Germany 28 June-1 July 2003]. PMID- 15125443 TI - Behold the power of a portfolio. PMID- 15125444 TI - Settling down. PMID- 15125445 TI - Professional and public attitudes toward incentives for organ donation. AB - The U.S. faces a widening gap between the need for, and the supply of, transplantable organs. The waiting list for transplants increased 150% in the past decade; last year, about 6,000 people died awaiting a transplant. This need has rekindled debate about the morality and feasibility of using incentives to encourage posthumous organ donation. This Issue Brief explores attitudes of the public and health professionals in the transplant community about using financial and nonfinancial incentives to increase the supply of cadaver organs for transplant. PMID- 15125446 TI - Abstracts of the 7th Annual Meeting of the Scientific Association of Swiss Radiation Oncology (SASRO) joint meeting with the Swiss Society of Radiobiology and Medical Physics (SSRMP). April 3-5, 2003, Geneva, Switzerland. PMID- 15125447 TI - Abstracts of the American Geriatrics Society Scientific Meeting. May 17-21, 2004, Las Vegas, Nevada, USA. PMID- 15125448 TI - Digestive Disease Week and the 105th Annual Meeting of the American Gastroenterological Association. May 15-20, 2004, New Orleans, Louisiana, USA. Abstracts. PMID- 15125449 TI - Orthopaedic proceedings: 2002, 2003. PMID- 15125450 TI - Sedation for endoscopic procedures in children. PMID- 15125451 TI - Medicare Savings Programs: helping your clients get and use the benefits. AB - Like those who are eligible for full Medicare and full Medicaid, people eligible for the Medicare Savings Programs are also referred to as dual eligibles or dual enrollees. And like their counterparts, they face special access and eligibility issues. In this second brief in our series on dual enrollees, we discuss issues that your clients may face as they seek to enroll in and use Medicare Savings Programs. PMID- 15125452 TI - From coast to coast: regional variations in the affordability of health care: findings from the Commonwealth Fund Biennial Health Insurance Survey. PMID- 15125453 TI - Prenatal care utilization and the implementation of prophylaxis to prevent perinatal HIV-1 transmission. AB - OBJECTIVES: To describe prenatal care utilization among women with HIV-1 in 4 US states, and to determine whether the adequacy of prenatal care utilization is associated with the implementation of prenatal, intrapartum, and postnatal HIV antiretroviral therapy (ARV). METHODS: Three-hundred three women completed a prenatal interview. Prenatal, labor and delivery, and infant medical records were reviewed. RESULTS: Thirty-nine percent of women did not receive adequate prenatal care; nearly one quarter of women did not begin care within the recommended timeframe, and approximately one-fifth of women received fewer than the recommended number of prenatal care visits from the time of entry into care until delivery. Those classified as less than adequate in terms of receipt of recommended visits were at increased risk for not receiving ARV during the prenatal care period and during labor and delivery, and were more likely to have had an infant subsequently diagnosed with HIV infection. CONCLUSION: Although women with HIV require adequate prenatal care for their own health as well as to improve perinatal outcomes, many are at risk for not receiving this care. Lower adherence to prenatal care appointments is an important risk factor for not receiving full HIV prophylactic regimens. PMID- 15125454 TI - A new method for classifying patterns of prenatal care utilization using cluster analysis. AB - OBJECTIVES: The objectives of this study were: to 1) define patterns of prenatal care utilization using cluster analysis, 2) describe two alternative cluster solutions and compare these groupings to the Adequacy of Prenatal Care Utilization Index (APNCU), 3) compare the cluster solutions and the APNCU with respect to maternal age and prematurity, and 4) discuss advantages and disadvantages of using cluster analysis to study prenatal care. METHODS: The study sample included 3544 women in the 1988 National Maternal and Infant Health Survey for whom complete prenatal care visit data were available. Clustering was carried out in two stages, first employing nearest centroid sorting (the k means method), a nonhierarchical approach, and then using Ward's Minimum Variance Method, a hierarchical clustering technique. RESULTS: Patterns of prenatal care defined by cluster analysis varied by timing of the first visit, total number of visits, and the rate of accumulation of visits, but this variation was different compared to that seen for the APNCU. While the cluster solutions and the APNCU identified a similar normative pattern of care, other patterns identified were quite different. In particular, the six-cluster solution differentiated among women who entered care at similar times, but accumulated visits at differing rates and experienced differing rates of preterm delivery. CONCLUSION: Cluster analysis is a new tool for studying prenatal care. Further studies are needed to refine the method and test whether the alternative perspective it provides will lead to new findings concerning the relationship of prenatal care and birth outcomes. PMID- 15125455 TI - Improving the accessibility of an analytic and technical skills MCH toolbox. AB - OBJECTIVES: To assess our analytic and technical skills website for accessibility and to make necessary corrections. METHODS: We used commercially available software (Jaws and LIFT for Dreamweaver) and an individual with visual impairments to evaluate our self-instructional, analytic, and technical tools. We identified problems in tables, images, multimedia content, PDF files, and links. RESULTS: We repaired the site by using LIFT to make appropriate modifications to the website and tools. CONCLUSIONS: Improving accessibility is advantageous for all Internet users. In addition to responding to legislative mandates, accessible web design creates pages that are often more readable, easier to navigate, and faster to download. Improving the accessibility of websites that incorporate charts and graphs strengthens the ability of all members of the MCH workforce to address the core functions. PMID- 15125456 TI - HIPAA transition: challenges of a multisite medical records validation study of maternally linked birth records. AB - Numerous researchers have expressed concern over the impacts on medical records availability of the newly effective Medical Information Privacy rule, as authorized by the Health Insurance Portability and Accountability Act (HIPAA). The increased costs associated with compliance with the rule, and the increased potential for financial liability, raises the possibility that hospitals may be less likely to participate in such research, resulting in a decrease of the validity of multisite studies designed to represent an entire population. Our multisite medical record validation study, designed to assess the accuracy of maternally linked birth records, provides an overview of a number of HIPAA implementation challenges. We found that the new HIPAA rule presents new challenges for those who rely on the release of medical record information for epidemiologic research. At the very minimum, increased compliance costs associated with human subjects protection and increased administrative burden for researchers would seem to be inevitable as medical institutions address the requirements of the new HIPAA rule by instituting more complex and thus more cumbersome procedures. Researchers should anticipate increased costs and plan accordingly when budgeting for human subjects review processes. PMID- 15125457 TI - 2003 National MCH Epidemiology Awards: recognizing excellence. PMID- 15125458 TI - Challenges facing MCH leadership: Martha May Eliot Award Commentary, 2003. PMID- 15125459 TI - A health care needs assessment of federal inmates in Canada. PMID- 15125460 TI - Helicobacter pylori test-and-treat strategy in the management of dyspepsia in primary care: an overview. PMID- 15125461 TI - Protective role of vanilloid receptor type 1 in HCl-induced gastric mucosal lesions in rats. AB - BACKGROUND: Effects of vanilloid-receptor agonists and antagonists on HCl-induced gastric lesions in rats were investigated to elucidate the role of vanilloid receptor type 1 (VR1) in gastric mucosal defense mechanisms. METHODS: Gastric lesions in rats were evaluated after intragastric administration of 0.6 N HCl. The localization of VR1 in the stomach was investigated immunohistochemically. RESULTS: Intragastric administration of capsaicin inhibited the formation of gastric lesions in a dose-dependent manner (0.1-2.5 mg/kg). The functional VR1 antagonists ruthenium red and capsazepine markedly aggravated HCl-induced gastric lesions in rats. The gastroprotective effect of capsaicin was attenuated by ruthenium red or capsazepine. It is reported that resiniferatoxin, [6]-gingerol and lafutidine are compounds that activate VR1 and/or capsaicin-sensitive afferent neurons. These compounds significantly inhibited the formation of HCl induced gastric lesions, and their gastroprotective effects were inhibited by treatment with ruthenium red. The immunohistochemical studies revealed that nerve fibers expressing VR1 exist along gastric glands in the mucosa, around blood vessels in the submucosa, in the myenteric plexus, and in the smooth muscle layers, especially the circular muscle layer. CONCLUSION: The results of this study suggest that VR1 plays a protective role in the gastric defensive mechanism in rats. PMID- 15125462 TI - Differential roles of interleukin-1beta and interleukin-8 in neutrophil transendothelial migration in patients with Helicobacter pylori infection. AB - BACKGROUND: Little information is currently available on the contribution of locally generated inflammatory and chemotactic cytokines to endothelial cell activation and subsequent neutrophil transendothelial migration in patients with Helicobacter pylori (H. pylori)-associated gastritis. METHODS: The contents of interleukin (IL)-1beta and IL-8 in the organ culture supernatants of antral mucosal tissues were measured with an enzyme-linked immunosorbent assay. The effects of the endogenous IL-1beta and IL-8 in mucosal tissues on neutrophil adherence and transendothelial migration were investigated using an experimental model of human umbilical vein endothelial cells (HUVEC). RESULTS: The contents of IL-1beta and IL-8 in organ cultures of antral mucosal tissues were significantly higher in patients with H. pylori infection than in those without infection. The organ culture supernatants from H. pylori-positive patients induced the expression of intercellular adhesion molecule-1 mRNA in HUVEC with increased binding of neutrophils, and these stimulatory effects were inhibited when HUVEC were pretreated with a nuclear factor-kappaB inhibitor, MG-132. Moreover, neutrophil adherence to HUVEC induced by the supernatants decreased after preincubation with neutralizing anti-IL-1beta antibody. As compared with the supernatants from H. pylori-negative patients, the samples from H. pylori positive patients exhibited a significantly higher chemotactic activity for neutrophils, which was inhibited almost completely by preincubation of the supernatants with anti-IL-8 antibody. CONCLUSIONS: Locally generated IL-1beta and IL-8 could coordinate with each other during the process of neutrophil infiltration into the gastric mucosa in patients with H. pylori infection. PMID- 15125463 TI - Sonographic detection of longitudinal ulcers in Crohn disease. AB - BACKGROUND: The purpose of this study was to investigate the characteristic sonographic features, such as focal disappearance of the wall stratification sign (FD sign), of longitudinal ulcers in patients with Crohn disease (CD). METHODS: A total of 545 sonographic examinations of patients with Crohn disease (n = 166), ulcerative colitis (n = 196), bacterial colitis (n = 78), ischemic colitis (n = 63), pseudomembranous colitis (n = 32), Behcet's disease (n = 7), and collagenous colitis (n = 3) were extracted. The sonographic findings were compared with those of a barium contrast study, colonoscopy, and resected specimens. In transverse views of the bowel segment, wall stratification was investigated. The FD sign was defined as the disappearance of the layered structure observed in a single portion of the bowel circumference. Prevalence of the FD sign was investigated for each disease. RESULTS: Eighty lesions (76 CD, 1 ulcerative colitis, 2 ischemic colitis, 1 Behcet's disease) with active single longitudinal ulcers were detected by barium contrast study and endoscopy. Among them, the FD sign was detected by ultrasonography (US) in 73 lesions (91.3%). No lesions with the FD sign were found by US without radiological or endoscopic findings of longitudinal ulcers. An in vitro study (water-immersion method) of resected specimens revealed that the FD sign reflected the focal destruction of wall stratification caused by the deep longitudinal ulceration. CONCLUSIONS: Ultrasonographic findings of FD sign are correlated with the existence of deep longitudinal ulcers, which are most frequently found in CD. US is a useful diagnostic modality for detecting longitudinal ulcers in patients with CD. PMID- 15125464 TI - Does the 'test-and-treat' strategy work in primary health care for management of uninvestigated dyspepsia? A prospective two-year follow-up study of 1552 patients. AB - BACKGROUND: Although the 'test-and-treat' strategy is suggested as first-line therapy for uninvestigated dyspepsia, no large-scale studies in a real-life setting are available. METHODS: 1552 dyspeptic patients aged between 25 and 60 with no alarm symptoms were recruited to the study. After screening with a 13C urea breath test, they were randomized into three treatment arms: Helicobacter pylori-positive either to eradication therapy with OAM (omeprazole, amoxycillin and metronidazole) (Hp+/erad) or omeprazole 20 mg daily (Hp+/ome) for 10 days, whereas H. pylori-negative patients (Hp-/ome) were treated with 20 mg omeprazole for 10 days. Gastrointestinal symptoms were registered at baseline at 1 and 2 years on the Gastrointestinal Symptom Rating Scale (GSRS) and quality of life with the Psychological General Well-Being index (PGWB). Additional visits, referrals for and number of endoscopies and their findings were registered during the 2 years' follow-up. RESULTS: Of the 1552 patients, 583 were H. pylori positive (37.6%), and 288 of these were randomized for omeprazole and 295 to OAM. The Hp-/ome group had fewer general practitioner (GP) contacts (P<0.0001) than the H. pylori-positive groups. Eradication therapy significantly improved general well-being and reduced upper gastrointestinal symptoms: abdominal pain (P=0.0001), heartburn (P=0.0061), acid regurgitation (P=0.003), hunger pain (P=0.009), especially in Hp+/erad. Peptic ulcer was found in 6.2%, 1.0%, 0.2% in Hp+/ome, Hp-+/erad and Hp-/ome, respectively (P=0.0007). Only 3 patients (1.0%) developed peptic ulcers in Hp-+/erad, all eradication failures. CONCLUSIONS: In uninvestigated dyspepsia, a negative test result for H. pylori reduces the number of GP contacts and endoscopy referrals compared to H. pylori-positive regardless of eradication therapy. Applied in real life, the test-and-treat strategy failed to reduce the number of endoscopies, but significantly reduced peptic ulcer disease and improved dyspeptic symptoms and quality of life. PMID- 15125465 TI - The burden cost of French patients suffering from irritable bowel syndrome. AB - BACKGROUND: The impact of irritable bowel syndrome (IBS) on health-care resource use in France is evaluated, and explanatory variables determined. METHODS: A questionnaire comprising socio-demographic characteristics, symptoms, consumption of resources, quality of life and impact of IBS on productivity was administered by telephone to a sample of 253 French adults with IBS recruited from the general population, and diagnosed with IBS using several well-known diagnostic criteria. The medical costs were estimated on a monthly basis and included medication(s), physicians' consultations, investigations and hospitalizations. RESULTS: Mean age was 48.3 years and 75% of subjects were women (192). Thirty-six percent of subjects had suffered from IBS for more than 10 years; 77% had consulted a general practitioner and 43% a gastroenterologist. Twenty-nine percent of subjects had undergone an investigation and 25% reported hospitalization; 61% of patients reported that they were taking medication. The average monthly medical costs was 71.8 euros (95% CI = [57.6-86.0]) with an asymmetric distribution (median = 28.1 euros) because of a high proportion of subjects (27%) who reported receiving no care at all. The two principal cost components were investigations (39%), and hospitalizations (22%). The highest medical costs were associated with subjects who were very elderly or suffered from severe symptoms (very severe pain), and were correlated with the lowest quality of life scores. CONCLUSION: IBS has a major impact on resource consumption and the productivity of patients. Determination of the variables to explain medical costs showed that advanced age, severe pain and deterioration in quality of life could be predictive of high medical costs. PMID- 15125466 TI - Vomiting and gastric electrical dysrhythmia in dogs. AB - BACKGROUND: The correlation between gastric myoelectrical activity (GMA) and gastrointestinal symptoms such as nausea and vomiting is poorly understood. The aim of this study was to assess the association of GMA with vomiting induced by retrograde gastric electrical stimulation or duodenal balloon distention. METHODS: Ten dogs were involved in this study. Vomiting was induced by retrograde gastric electrical stimulation in 6 dogs and by duodenal balloon distention in 4 dogs. Computerized spectral analysis and visual analysis were applied to detect the GMA change during various periods before and after vomiting. RESULTS: Gastric dysrhythmia preceded vomiting but was of brief duration. The major pattern of dysrhythmia immediately before vomiting was tachyarrhythmia and gastric slow wave was completely uncoupled before vomiting. Gastric dysrhythmia and slow wave uncoupling were also noticed immediately after vomiting but the dogs recovered quickly. The major pattern of dysrhythmia after vomiting was arrhythmia. GMA was normal during the periods other than 5 min before and during vomiting and 5 min after vomiting. CONCLUSIONS: Gastric dysrhythmia seems to be the cause of vomiting induced by retrograde gastric electrical stimulation or duodenal balloon distention. It is brief and characterized with tachyarrhythmia and uncoupling. PMID- 15125467 TI - Glucagon-like peptide-2 inhibits antral emptying in man, but is not as potent as glucagon-like peptide-1. AB - BACKGROUND: GLP-1 (glucagon-like peptide-1) and GLP-2 (glucagon-like peptide-2) are released in equimolar amounts in response to meal ingestion. GLP-1 inhibits gastric emptying and reduces postprandial gastric and exocrine pancreatic secretion and may play a physiological regulatory role in controlling appetite and energy intake in humans. The role of GLP-2 is more uncertain. Based on the results of animal studies, it has been suggested that GLP-2 may induce intestinal epithelial growth and inhibit gastric motility. The aim of this study was to determine to what extent GLP-2 alone or together with GLP-1 inhibits gastric emptying and the sensation of hunger in man. METHODS: Eight healthy volunteers were tested in a double-blind, placebo-controlled fashion. Antral emptying of a liquid meal and hunger ratings were determined using ultrasound technology and visual analogue scales scoring during infusions of saline, GLP-2 (0.5, and 1.0 pmol kg body wt(-1) min(-1)), GLP-1 (0.5 pmol kg body wt(-1) min(-1)) or GLP-1 and GLP-2 (0.5 pmol kg body wt(-1) min(-1)). RESULTS: The GLP-2 infusions resulted in a dose-dependent increase in antral emptying time (35%; ns and 75%; P = 0.049) compared to saline, but GLP-2 was less potent than GLP-1, which increased the antral emptying time by 192% (P < 0.001). Addition of GLP-2 to the GLP-1 infusion did not alter the antral emptying time compared with GLP-1 alone. The GLP-1 infusion decreased the sensation of hunger compared with saline (P = 0.023), whereas the two GLP-2 infusions had no significant effect. Addition of GLP-2 to the GLP-1 infusion did not decrease the sensation of hunger further. CONCLUSIONS: Both GLP-1 and GLP-2 inhibit antral emptying in man, but GLP-1 is more potent. PMID- 15125468 TI - Coeliac disease in The Netherlands. AB - BACKGROUND: The prevalence of adult coeliac disease in The Netherlands was studied in the Dutch Coeliac Disease Society and in blood donors but not in the general population. We therefore studied the prevalence of recognized and unrecognized coeliac disease in a large cohort, representative of the adult Dutch general population. Blood samples were available for anonymous research, as well as data on dietary habits, self-reported physical characteristics, health problems, quality of life and socio-economic circumstances. METHODS: Subjects included 50,760 individuals who had previously participated in two large population-based studies on health status in relation to lifestyle factors. Recognized coeliac disease was studied in all subjects by identification of self reported adherence to a gluten-free diet and subsequent confirmation of the diagnosis of coeliac disease. Unrecognized coeliac disease was studied in a random sample of 1440 out of the 50,760 subjects through serologic screening and human lymphocyte antigen (HLA) typing. RESULTS: The prevalence of recognized coeliac disease was 0.016% (95% confidence interval 0.008-0.031) and of unrecognized coeliac disease 0.35% (95% confidence interval 0.15-0.81). Menarcheal age was higher in women with recognized coeliac disease than in women without coeliac disease. CONCLUSIONS: The prevalence of adult recognized coeliac disease in The Netherlands is one of the lowest in Europe, while the prevalence of unrecognized coeliac disease is comparable with that in other European countries. Adult coeliac disease is strongly under diagnosed in The Netherlands. The higher menarcheal age in women with recognized coeliac disease may be explained by diagnostic delay. PMID- 15125469 TI - Health-related quality of life in patients with inflammatory bowel disease five years after the initial diagnosis. AB - BACKGROUND: Health-related quality of life (HRQOL) has become an important tool in evaluating patient satisfaction in inflammatory bowel disease (IBD). So far, few prospective follow-up studies have been done to identify variables that influence HRQOL. We aimed to identify demographic and clinical variables that influence HRQOL 5 years after diagnosis in patients with ulcerative colitis (UC) or Crohn disease (CD) included in a prospective follow-up study from 1990 to 1994 (the IBSEN study). METHODS: All patients completed the Inflammatory Bowel Disease Questionnaire (IBDQ), a disease-specific quality-of-life questionnaire translated into Norwegian and validated. We present data from 497 patients (328 UC patients and 169 CD patients, mean age 43.3 years, 48% female). The impact of age, gender, smoking, symptom severity, disease distribution, rheumatic symptoms and surgery on IBD patients' HRQOL was analysed. RESULTS: Women had a reduction in IBDQ total score of 10 points compared to men, CD patients had a reduction of 7.5 compared to UC patients. The patients with moderate/severe symptoms had a 50 points lower score than the patients without symptoms. The patients with rheumatic symptoms had a 10 points lower total score than the patients without these symptoms. All differences were statistically significant. The multiple regression analysis showed that symptom severity, rheumatic symptoms and female gender were the strongest predictors of reduction in HRQOL for both diagnosis groups. CONCLUSION: IBD symptoms, rheumatic symptoms and female gender have a significant influence on patients' HRQOL as measured by IBDQ. This was confirmed by the regression analysis. PMID- 15125470 TI - Sexuality in patients with ulcerative colitis before and after restorative proctocolectomy: a prospective study. AB - BACKGROUND: The purpose of this study was to assess sexuality in patients with ulcerative colitis (UC), before and one year after an ileal-pouch anal anastomosis (IPAA). METHODS: Group I comprised 14 medically treated patients (7 M) and Group II comprised 29 patients (18 M), operated with colectomy and ileostomy, with preservation of the rectum. Patients were interviewed about desire, excitement and general sexual satisfaction before and one year after the IPAA operation. RESULTS: Most of the patients had restarted sexual activity < or = 3 months after surgery. Sexual desire and desire for intercourse had not changed from before the IPAA operation. In Group I, one woman suffered from dyspareunia both before and after the IPAA. In Group II, three women had dyspareunia before the IPAA and four after surgery. Erection and ejaculation were normal in all men before the IPAA operation. After IPAA, one man in Group I and two men in Group II had loss of ejaculation. In Group 1, 10 out of 14 patients were satisfied with their sexual life both before and after the IPAA. In Group II the corresponding figures were 15 out of 28 before and 22 after IPAA surgery. CONCLUSIONS: A third of the patients stated that they were dissatisfied with their sexual life before IPAA. Although a few patients experienced sexual imperfections such as loss of ejaculation and dyspareunia, the majority of patients in both groups considered that the overall general satisfaction with their sexual life had normalized considerably after surgery. PMID- 15125471 TI - Lymph node metastasis as a significant prognostic factor in gastric cancer: a multiple logistic regression analysis. AB - BACKGROUND: In Japan, the standard treatment policy for all potentially curable patients with gastric cancer is radical resection including extensive lymphadenectomy. This treatment strategy has been used for both early and advanced gastric cancers, and substantial increases in survival time have been reported. In advanced gastric cancer, lymphatic spread is reported to be one of the most relevant prognostic factors for gastric cancer resected for cure. The purpose of this study was to determine the factors affecting lymph node involvement and to establish guidelines for the extent of lymph node dissection most appropriate for the treatment of gastric cancer. METHODS: The clinicopathological features of 926 patients with gastric cancer were reviewed. Information on the clinicopathological features was obtained from the database of gastric cancer at the Department of Gastroenterological Surgery, Sendai National Hospital. Univariate and multivariate analyses of data for patients with gastric cancer tumors were performed to evaluate the prognostic significance of clinicopathological features. The independent risk factors influencing lymph node metastasis were determined by multiple logistic regression analysis. RESULTS: The following clinicopathologic factors were found to be correlated with prognosis of gastric cancer: (1) macroscopic type, (2) depth of invasion, (3) cancer-stromal relationship, (4) histological growth pattern, (5) lymph node involvement, (6) lymphatic invasion, (7) vascular invasion and (8) tumor site. However, a multivariate analysis revealed that macroscopic type, depth of invasion, lymph node involvement and tumor site are independent risk factors for the prognosis of gastric cancer patients. Among these factors, the prognosis of patients with gastric cancer was most strongly influenced by lymph node involvement (odds ratio, 4.632). According to a multiple logistic regression model, depth of cancer invasion and lymphatic invasion was significantly correlated with lymph node metastases. CONCLUSIONS: Lymph node involvement has the strongest influence on the prognosis of gastric cancer. Among the clinicopathological factors, depth of invasion and microscopically lymphatic invasion are important factors in predicting lymph node metastases. Thus, the ability to perform gastrectomy with dissection of lymph nodes is a basic requirement for gastric cancer surgeons. PMID- 15125472 TI - Effect of celecoxib on renal function in cirrhotic patients with ascites. A pilot study. PMID- 15125473 TI - Studies on tissue expression of HCV proteins (NS3 and C) in chronic hepatitis C using the ImmunoMax technique. PMID- 15125474 TI - A patient with myelofibrosis complicated by refractory ascites and portal hypertension: to tips or not to tips? A case report with discussion of the mechanism of ascites formation. AB - In patients with myelofibrosis, clinically significant portal hypertension is known to be predominantly presinusoidal; however, the exact mechanisms are still controversial. The pathophysiology is particularly enigmatic in those patients without histological and angiographic evidence of significant intra- or extrahepatic obstruction to portal blood flow, respectively. Moreover, ascites formation has been reported in such cases, but in general is rare in presinusoidal portal hypertension. Here we present such a patient in which ascites developed even in the presence of unchanged serum protein levels (oncotic pressure) and was refractory to sodium restricted diet and high-dose diuretic treatment. A discussion on the parameters influencing fluid exchange and ascites formation particularly emphasizing the potential importance of the hyperdynamic circulation in this case is given. Finally, the patient was treated by implanting a transjugular intrahepatic shunt (TIPS), exerting a diuretic effect sufficient enough to avoid re-formation of ascites for several months. However, ascites re accumulated potentially due to the appearance of ectopic peritoneal myeloid metaplasia and the patient died soon afterwards. In conclusion, TIPS may be considered as rescue management for refractory ascites secondary to portal hypertension, but caution in respect to the presence and/or development of peritoneal or other ectopic haematopoesis has to be taken. PMID- 15125475 TI - Successful treatment of pancreatobiliary fistula by endoscopic stenting. AB - An unusual pancreatobiliary fistula occurred as a complication of chronic pancreatitis. Endoscopic papillotomy was performed and a plastic endoprosthesis was inserted into the main pancreatic duct. The pancreatic stenting led to the closure of the fistula and no additional surgical treatment was necessary. PMID- 15125476 TI - A novel approach to a patient with Crohn disease and a high stoma output: a missed opportunity? AB - The development of stricturing or penetrating behaviour in Crohn disease is influenced by disease location, clinical activity of the disease and smoking habit. The management of inflammatory bowel disease remains unsatisfactory, and the influence of current therapies--immunosuppressants, and especially the novel biological treatment now commonly used in Crohn disease--remains largely unknown. A relatively smaller number of patients receive dietary treatment as well. Physicians' scepticism, under-resourced dietetic departments and patient compliance are the main concerns in considering enteral nutrition. The management of high stomal output in Crohn disease is even more challenging. We report a case of active Crohn disease with high stoma output treated successfully solely with enteral feeding without the need for surgical intervention or administration of potent pharmacotherapy. PMID- 15125477 TI - Sarcoidosis in untreated chronic hepatitis C virus infection. AB - Chronic hepatitis C has been implicated in the pathogenesis of sarcoidosis in several cases of patients treated with interferon-alpha. On the other hand, only in a few cases previously has a possible link between sarcoidosis and untreated chronic hepatitis C virus infection been demonstrated. We report on a patient with chronic hepatitis C who developed cutaneous sarcoidosis without prior interferon-alpha treatment. We hypothesize that viral persistence seen in chronic hepatitis C virus infection might be one of the potential factors that trigger cellular immune response in granulomatous reactions as seen in sarcoidosis, in genetically predisposed patients. PMID- 15125478 TI - Tegaserod--merely a laxative? PMID- 15125479 TI - Specificity of anti-sp100 antibody for primary biliary cirrhosis. PMID- 15125480 TI - Anti-sp100 antibodies in primary biliary cirrhosis. PMID- 15125481 TI - Inflammatory status and endothelial function in asymptomatic and symptomatic peripheral arterial disease. AB - Peripheral arterial disease (PAD) is a predictor of cardiovascular risk. However, it is unknown whether PAD severity influences inflammatory status and endothelial function, which play a major role in atherosclerosis. Accordingly, we measured brachial artery flow-mediated dilation (FMD), and plasma levels of several inflammatory markers in 15 control subjects, and 19 asymptomatic and 19 symptomatic PAD patients. Each symptomatic patient was matched to an asymptomatic patient for age, sex, risk factors, presence of cardiovascular disease, and pharmacological treatments. Asymptomatic patients had similar inflammatory profiles as controls, but lower median FMD (11.7% vs 8.5%, p < 0.01). Compared with asymptomatic patients, symptomatic patients had higher median C-reactive protein (1.5 mg/l vs 6.0 mg/l, p < 0.05) and interleukine-6 (1.5 pg/ml vs 3.5 pg/ml, p < 0.05), and lower FMD (8.5% vs 5.1%, p < 0.01). In the 38 PAD patients, the ankle/brachial pressure index correlated positively with FMD (p < 0.01), and negatively with C-reactive protein (p < 0.05), soluble intercellular adhesion molecule-1 (p < 0.05) and soluble vascular cell adhesion molecule-1 (p < 0.05). Thus, in PAD, endothelial function and inflammatory status are related to the severity of the circulatory impairment. This finding may contribute to the explanation of the increasingly poor prognosis with increased PAD severity. PMID- 15125482 TI - Lower extremity peripheral arterial disease in hospitalized patients with coronary artery disease. AB - The prevalence of coronary artery disease (CAD) in patients with peripheral arterial disease (PAD) has been well defined. However, the prevalence of PAD in hospitalized patients with CAD has not been defined. The ankle-brachial index (ABI) is a useful non-invasive tool to screen for PAD. The aim of our study was to assess the prevalence of PAD in hospitalized patients with CAD by measuring the ABI. The study was conducted at the University of Wisconsin Hospital and Clinics inpatient Cardiovascular Medicine Service. Medically stable patients with CAD were invited to participate prior to hospital discharge. Data regarding cardiovascular risk factors, history of previous PAD, physical examination, and ABI were collected. An ABI less than or equal to 0.9 or a history of previous lower extremity vascular invention was considered to be indicative of significant PAD. A total of 100 patients (66 men and 34 women) were recruited. Forty patients were found to have PAD (mean ABI in non-revascularized patients with PAD = 0.67). By measuring the ABI, 37 (25 men) were positive for PAD and three had an ABI corrected with previous revascularization. Of these patients, 21 (52.5%) had previously documented PAD. Patients with PAD were older (p = 0.003), had a greater smoking history (p = 0.002), were more likely to have diabetes (p = 0.012), hypertension (p = 0.013) and a trend towards more dyslipidemia (p = 0.055). In conclusion, hospitalized patients with CAD are likely to have concomitant PAD. Risk factors for PAD in this patient population include advanced age, history of smoking, diabetes, hypertension, dyslipidemia and abnormal pulse examination. Identification of patients with PAD by measuring the ankle-brachial index is easily done. PMID- 15125483 TI - Ethnic differences in peripheral arterial disease in the NHLBI Genetic Epidemiology Network of Arteriopathy (GENOA) study. AB - Few studies have investigated whether ethnic groups differ in the prevalence of peripheral arterial disease (PAD). We compared the distribution of the ankle brachial index (ABI), a measure of PAD, between African Americans and non Hispanic white individuals. Subjects (n = 931) belonged to the Genetic Epidemiology Network of Arteriopathy (GENOA) study, a community-based study of hypertensive sibships, and included 453 African Americans from Jackson, Mississipi (mean age 72 +/- 6 years, 69% women) and 478 non-Hispanic white individuals from Rochester, Minnesota (mean age 58 +/- 7 years, 64% women). ABI was determined at two sites in each lower extremity and the lowest of four indices was used in the analyses. PAD was defined as an ABI of < or = 0.95. Information about conventional risk factors was derived from interviews and from blood samples drawn at the study visit. The prevalence of diabetes and hypertension was significantly higher in African Americans than in non-Hispanic white individuals. After adjusting for age, African American subjects had a lower mean ABI (women 0.97 vs 1.04, p < 0.001; men 0.96 vs 1.12, p < 0.001) and a greater prevalence of PAD (women 34% vs 22%, p = 0.010; men 33% vs 11%, p < 0.001) than their non-Hispanic white counterparts. In multiple regression analyses, African American ethnicity was a predictor of a lower ABI and the presence of PAD in each sex after adjusting for age and other conventional risk factors. In conclusion, the lower ABI and greater prevalence of PAD in African Americans than in non-Hispanic white individuals is not explained by differences in conventional risk factors. Identifying additional 'novel' risk factors that account for the ethnic differences in PAD is an important next step towards understanding why such differences exist and developing more effective strategies to reduce the burden of PAD. PMID- 15125484 TI - Effect of sildenafil on arterial stiffness and wave reflection. AB - While sildenafil (Viagra) is widely prescribed for erectile dysfunction, its effect on arterial function is not established. The elastic properties of the aorta, as well as the magnitude and timing of wave reflection, are important factors for efficient performance of the cardiovascular system and have been identified as prognosticators of cardiovascular risk. A total of 24 subjects with coronary artery disease, of whom 14 were hypertensives, aged 69 +/- 8 years, were studied in a randomized, placebo-controlled, double-blind, cross-over design. Measurements lasted for 3 h after the sildenafil intake (50 mg, p.o.) or placebo. Aortic elastic properties were evaluated with carotid-femoral pulse wave velocity; wave reflection was evaluated with augmentation index and augmented pressure of the aortic pressure waveform. Pulse wave velocity decreased significantly (by 0.65 m/s, p = 0.005), denoting a decrease in aortic stiffness. Augmentation index and augmented pressure decreased significantly (by 4.47% absolute and by 4.01 mmHg; p < 0.001 and p = 0.001, respectively), denoting a decreased effect of wave reflection from the periphery. Aortic pulse pressure decreased significantly (by 6.74 mmHg, p < 0.05). An active effect of the drug on aortic wall appears to contribute to the decrease in pulse wave velocity, although other mechanisms such as a decrease of blood pressure and autonomic reflexes could also have contributed. The effect of sildenafil lasted throughout the study (3 h), being evident 30 min after drug intake. In conclusion, this study shows, for the first time, that sildenafil has a favorable effect on aortic stiffness and wave reflection in patients with coronary artery disease. This finding may have important implications for cardiovascular performance and exercise capacity during intercourse. PMID- 15125485 TI - Peripheral veins: influence of gender, body mass index, age and varicose veins on cross-sectional area. AB - To investigate changes in the size of the deep and superficial venous systems associated with gender, age, body mass index and varicose veins, changes to the cross-sectional area of the femoral and the long saphenous veins were analysed in the Duesseldorf/Essen civil servant study population. Between December 1989 and July 1993 a total of 9935 employees were recruited; 9261 were then evaluated for this analysis. Diameters of the long saphenous and femoral veins were determined 2-3 cm distal to the confluence in lying (after 15 min rest) and standing (after 5 min) positions. Cross-sectional areas (CSA) were calculated. A total of 63% of all people were assigned to CEAP (clinical, etiological, anatomical pathophysiological) class 0, 27% to class 1, 8.5% to class 2, while 1.5% belonged to higher CEAP classes. In people without varicose veins (CEAP class 0) the CSA of the femoral and long saphenous veins were smaller in females than in males. In people with a normal body mass index (BMI) (20-25) the mean CSA of the femoral and long saphenous veins in a standing position was similar from the third up to the sixth decade of life. The volume increase due to a standing position expressed as the absolute increase in CSA of the femoral and long saphenous veins was not age-related, either. The relative volume increase expressed as a ratio remained unchanged with age. There was a strong relationship between the CSA of both veins and increasing BMI. In a lying position, the CSA of the femoral and long saphenous veins increased only slightly with increasing CEAP classes. In a standing position, the CSA of both veins increased even in CEAP class 1 (p < 0.001). In a stepwise multivariate regression analysis, the CSA of both veins in a standing position was not age-related but associated with BMI, CEAP classes and gender. The absolute increase in CSA was influenced by all four variables, but BMI and gender were most important. In Conclusion, this study shows that aging is not necessarily associated with an increase in venous CSA of the deep and superficial venous system. BMI is the most important determinant for an increase in CSA in standing position. Varicosity of the superficial venous system is always associated with similar changes in the deep venous system. PMID- 15125486 TI - Syncope in patients with pulmonary embolism: comparison between patients with syncope as the presenting symptom of pulmonary embolism and patients with pulmonary embolism without syncope. AB - Syncope as an initial presentation of pulmonary embolism occurs in 10% of patients. We compared clinical and instrumental parameters in patients with syncope as the presenting symptom of pulmonary embolism and in patients with documented pulmonary embolism without syncope. Seventy patients with the diagnosis of pulmonary embolism and apparently stable clinical conditions were evaluated. They were divided in two groups: 10 patients with syncope as the presenting symptom of pulmonary embolism (group 1) and 60 patients without syncope (group 2). Patients with syncope showed a more pronounced tendency to present with main pulmonary artery embolus than patients without syncope (contingency coefficient = 0.301, p < 0.04; one-tailed). However, despite the evidence that patients with syncope have significant reductions in systolic and/or diastolic blood pressure, shock was not observed in any patient. In no case was thrombolytic treatment given and all patients received standard anticoagulation with unfractioned heparin and oral anticoagulant. We suggest that syncope in the setting of non-massive pulmonary embolism may be due to vaso-vagal mechanism that can lead to a reduction of arterial blood pressure when central artery thrombosis is involved. PMID- 15125487 TI - Effect of alcohol on endothelial function in healthy subjects. AB - While studies have shown that red wine improves endothelial function, the effect of pure alcohol, the common denominator of all alcoholic beverages, on endothelial function has not been defined. The effect of oral intake of alcohol (one ounce--a dose corresponding to two drink equivalents) on flow-mediated dilatation of the brachial artery was studied in 12 healthy subjects using high resolution ultrasonography according to a randomized, sham procedure-controlled, single-blind protocol. Alcohol produced a significant vasodilatation of the brachial artery at resting conditions. Furthermore, it led to a significant increase of the brachial artery diameter at reactive hyperemia; however, the percentage flow-mediated dilatation did not change. This study shows, for the first time, that acute moderate intake of alcohol has a potent dilating effect on large arteries both at resting and at reactive hyperemic conditions, without affecting flow-mediated dilatation as a percentage. This may be useful for explaining the divergent effect of different alcoholic beverages on endothelial function and provides valuable insight into the mechanisms involved in the vascular effects of alcohol. PMID- 15125488 TI - The pivotal role of matrix metalloproteinases in the development of human abdominal aortic aneurysms. AB - Abdominal aortic aneurysms (AAAs) represent a chronic degenerative condition and impart the risk of a life-threatening episode of rupture. Chronic inflammation and destructive remodeling of the extracellular matrix of the aortic wall constitute trademarks of this entity. Multiple studies have implicated a group of locally produced matrix endopeptidases-the matrix metalloproteinases (MMPs)-as the main culprits of this process. For this reason, extensive research on the identification of the role of these enzymes, as well as possible alternative pharmacological treatments of AAAs, has taken place during the last few years. The exact role of the several members of the group of metalloproteinases has already been discovered, and conservative therapeutic strategies oriented towards these agents have been suggested, but a definite treatment plan is still a controversial topic. The possible role of a genetic predisposition to AAAs is another crucial topic that remains to be determined, as it would render the confrontation of this condition much more efficient. PMID- 15125489 TI - The potential role of angiotensin-converting enzyme inhibition in peripheral arterial disease. AB - Peripheral arterial disease (PAD) is associated with significant morbidity and mortality, and yet remains under-recognized and under-treated. Atherosclerosis is the most common cause of lower extremity PAD and pharmacological interventions that alter this central pathogenic role of atherosclerosis may alter the natural history of PAD. There is growing evidence that the renin-angiotensin system (RAS) is a significant mediator of this disease process and that treatment with angiotensin-converting enzyme (ACE) inhibitors is associated with vasculoprotective effects that are independent of the antihypertensive properties of these agents. Numerous lines of evidence suggest that ACE inhibitors directly inhibit the atherosclerotic process and improve vascular endothelial function. In patients with PAD, ACE inhibitors have been shown to improve peripheral circulation as measured by peripheral arterial blood pressure and by increases in peripheral blood flow. Preliminary evidence suggests that ACE inhibitors might improve clinical symptoms in patients with PAD. Recent evidence has confirmed that ACE inhibition is associated with a decrease in morbidity and mortality in patients with arterial disease without left ventricular dysfunction; this benefit was at least as great for the subset of patients with PAD. Overall, these data support a significant role for the RAS in the pathogenesis of all atherosclerotic diseases (including PAD) and suggest that the benefit is independent of the blood pressure lowering properties of these agents. These studies suggest that ACE inhibitor therapy should be considered in the routine management of individuals with PAD, regardless of whether they have hypertension or left ventricular dysfunction. PMID- 15125490 TI - Images in vascular medicine. Aortic dissection or extra-aortic artefact? PMID- 15125491 TI - Images in vascular medicine. Diagnosis of giant cell arteritis with PET/CT. PMID- 15125492 TI - Images in vascular medicine. Angiosarcoma of superior vena cava with extension into right atrium assessed by MD-CT and MRI. PMID- 15125493 TI - Images in vascular medicine. Distal abdominal aortic occlusion. PMID- 15125494 TI - Images in vascular ultrasound. Stenosis of the innominate or left common carotid artery origin diagnosed on carotid ultrasound examination. PMID- 15125495 TI - Quantifying effect of statins on low density lipoprotein cholesterol, ischaemic heart disease, and stroke: systematic review and meta-analysis. Law MR, Wald NJ, Rudnicka AR. BMJ 2003; 326: 1407-408. PMID- 15125496 TI - A conference report. The second Siena International Conference on animal models of chronic obstructive pulmonary disease. PMID- 15125497 TI - Lupus erythematosus induced by medications, ultraviolet radiation, and other exogenous agents: a review, with special focus on the development of subacute cutaneous lupus erythematosus in a genetically predisposed individual. AB - Exogenous agents implicated in or suspected of precipitating subacute cutaneous lupus erythematosus (SCLE) and lupus erythematosus (LE) are reviewed. An illustrative case of environmentally induced SCLE is presented. A previously healthy 30-year male homozygous for the tumor necrosis factor-alpha (TNF-alpha) 308. A promoter allele developed SCLE after spending several hours removing fertilizer- and pesticide-containing hay from an agricultural barn in the springtime. The cutaneous eruption soon resolved, only to reappear 3 weeks later on the day the patient re-entered the barn. An environmental agent present in the barn, coupled with springtime ultraviolet light, likely triggered the disease in this immunogenetically susceptible individual. PMID- 15125498 TI - Neutrophilic dermatosis of the hands: four new cases and review of the literature. AB - BACKGROUND: Isolated or predominantly hand involvement in Sweet's syndrome, pyoderma gangrenosum, or pustular vasculitis is a rare presentation in the spectrum of neutrophilic dermatoses and is often associated with an occult malignancy or other systemic inflammatory disorder. When these disorders occur on the hands, they are often clinically indistinguishable, but they can sometimes be separated histologically by the presence of papillary dermal edema (Sweet's syndrome), ulceration and necrosis (pyoderma gangrenosum), or vasculitis (pustular vasculitis). These distinctions may be arbitrary, however, and reflect differences in the temporal course of the disease and in the degree of inflammation at the time of biopsy. METHODS: We report four cases of neutrophilic dermatosis affecting the hands and a review of the literature for similar cases. RESULTS: Of the four patients presented, two had associated carcinomas and one had myelodysplasia in transition to leukemia. The cutaneous symptoms preceded the finding of an occult malignancy. Thirty-two reported cases of neutrophilic dermatoses presenting on the hands showed clinicopathologic features similar to those in our series. Taken together, the mean age at diagnosis was 60.5 years, and 58% of the patients were female. Twenty-five per cent (nine patients) also had myelodysplasia or leukemia, 14% (five patients) ulcerative colitis, 6% (two patients) carcinoma, 6% (two patients) Crohn's disease, and 6% (two patients) seropositive arthritis. CONCLUSIONS: These cases illustrate the importance of recognizing that neutrophilic dermatoses may present uniquely or predominantly on the hands. This presentation is distinctive, and prompt diagnosis may prevent unnecessary medical or surgical therapy and may lead to the earlier diagnosis and treatment of an associated malignancy or other systemic disorder. PMID- 15125499 TI - Propionibacterium acnes biotypes and susceptibility to minocycline and Keigai rengyo-to. AB - BACKGROUND: Propionibacterium acnes is the predominant organism in acne lesions, but the sensitivity of different biotypes of P. acnes to therapeutic agents has seldom been reported. METHODS: To characterize biotypes of P. acnes and to measure the effects of Keigai-rengyo-to (KRT) and minocycline (MINO) on clinical P. acnes isolates. RESULTS: Propionibacterium acnes biotype III (BIII) is the most common form of identified acne lesion, followed by P. acnes biotype I. BIII was isolated from mild, moderate and severe severity and the average lipase activity of BIII was higher than that of Biotypes I, II, IV and V. No significant differences in the decrease of free fatty acid production elicited by KRT or by MINO were found between BIII and the other biotypes. The degree of decreased butyric acid production was greater than that of propionic acid production in the medium supplemented with MINO. The percent decrease of butyric acid production elicited by 1 mg/mL of KRT was the same as that elicited by 0.1 microg/mL of MINO. Among biotypes of P. acnes, the minimal inhibitory concentrations of agents tested were generally higher in erythritol-positive biotypes than in erythritol negative biotypes. CONCLUSION: The high frequency of BIII might be responsible for the severity of acne in patients. It seems that if the same concentrations of MINO and KRT are used, the antilipase activity of MINO is stronger than that of KRT. Minocycline also has a direct anti-lipase activity against P. acnes. The mechanism underlying the influence of erythritol on the susceptibility of P. acnes to these agents remains unknown. PMID- 15125500 TI - Clinicopathologic features of ulcerative-atrophic sarcoidosis. AB - BACKGROUND: Sarcoidosis is a chronic granulomatous disease of unknown etiology. Cutaneous disease is common and includes two clinicopathologic categories: granulomatous infiltration or a reactive phenomenon. In the granulomatous infiltrative group, clinical manifestations can be variable. Ulcers in sarcoidosis are uncommonly recognized and have been categorized previously under the rubric of atrophic, necrobiosis-like, or ulcerative sarcoidosis. PATIENTS AND METHODS: We evaluated retrospectively sarcoidosis patients presenting to the Johns Hopkins Department of Dermatology between June 1989 and May 2002. Multiple skin biopsies were performed for histopathologic evaluation. Investigation for extracutaneous manifestations, including routine serologic assays, chest radiography, pulmonary function tests, electrocardiogram, and angiotensin converting enzyme level, and referral for ophthalmologic examination were performed in all patients. RESULTS: Of 147 consecutive patients presenting with cutaneous sarcoidosis, seven demonstrated ulcerative-atrophic sarcoidosis lesions. All patients were African-American (five females and two males). All patients had ulcers surrounded by atrophic necrobiosis lipoidica-like plaques on the pretibial areas. All patients had other mucocutaneous manifestations of sarcoidosis, with the majority having evidence of internal disease. Combined immunosuppressive and immunomodulatory therapy was effective in controlling the cutaneous manifestations of all patients with ulcerative sarcoidosis. CONCLUSIONS: The ulcerative variant is a poorly defined subset of cutaneous sarcoidosis. Trauma, superimposed on atrophic plaques, appears to be the principal mechanism of this rare variant of cutaneous sarcoidosis. PMID- 15125501 TI - Diagnosis of sports-related dermatoses. PMID- 15125502 TI - Localized cutaneous leishmaniasis imported into Paris: a review of 39 cases. AB - BACKGROUND: Localized cutaneous leishmaniasis (LCL) is a common cause of dermatosis in travelers returning from the tropics. We describe the epidemiological, clinical, and biological aspects and therapeutic outcome of imported LCL. METHODS: A retrospective study of all cases of LCL observed from 1992 to 2000 in our tropical disease unit. Diagnosis was based on direct examination of skin smear and/or culture with identification of subsequent subspecies. RESULTS: Thirty-nine cases (25 males, 14 females; median age: 38 years) were included: 35 French travelers and four foreign immigrants; 15 cases were acquired in the Old World and 24 cases in the New World. The patients presented to our department with a median of 60 days after return. Thirteen patients had already consulted general practitioners, and the diagnosis was missed in five cases (38%). Five clusters were identified. The median number of skin lesions was two per patient. Diagnosis was established by direct microscopic examination in 36 cases (92%). Thirty-five patients were assessable for first line treatment with antimonials (intramuscularly in 18, intralesionally in nine), intramuscular pentamidine isethionate or oral ketoconazole (four patients each). Twenty-five patients (71.4%) were cured. The remaining 10 patients were cured after one to three courses of other treatments. Overall adverse events occurred in 60% of the patients treated with antimonials and 37% of those treated with pentamidine. CONCLUSION: Imported LCL is still unrecognized by Western physicians. Clusters may be observed in groups of travelers. The therapeutic outcome is impaired by numerous but minor side-effects. PMID- 15125503 TI - Zosteriform skin metastases. PMID- 15125504 TI - Dr Karel Fleischmann: the story of an artist and physician in Ghetto Terezin. AB - Dr Karel Fleischmann (1897-1944) was a Jewish dermatologist and noted Czech artist. During World War II, Fleischmann was confined by the Nazis in Ghetto Terezin (Theresienstadt), which was located in the Protectorate of Bohemia and Moravia. While at Terezin, Fleischmann became a leader of the ghetto's health care system and administered programs that helped to dramatically reduce the high mortality rate of the Jewish population from disease. In his spare time, Fleischmann drew works of art that portrayed the daily life and suffering of the Jews in Ghetto Terezin. Although Fleischmann perished in the Auschwitz-Birkenau concentration camp, his artwork survived the war. This article pays tribute to Dr Karel Fleischmann by telling the story of his life and by citing selections from his art, poetry and prose. PMID- 15125505 TI - Mandibular alveolar bone necrosis after trigeminal herpes zoster. PMID- 15125506 TI - Case of leukoderma punctata after topical PUVA treatment. PMID- 15125507 TI - A pheochromocytoma presenting as generalized pigmentation. PMID- 15125508 TI - Vestibular papillae of the vulva. PMID- 15125509 TI - Linear and whorled nevoid hypermelanosis associated with developmental delay and generalized convulsions. PMID- 15125510 TI - Surgical option for nonmelanoma skin cancer. AB - BACKGROUND: Basal cell carcinoma (BCC) is one of the most frequent malignancies in the general population. Its best treatment option is the complete excision of the lesion. Mohs' micrographic surgery has demonstrated to be the surgical method with the highest cure rates, however, it is not available in many countries or institutions. METHODS: We propose, as a treatment option for high-risk BCC, surgical resection of the tumor with transoperatory histological examination with the hematoxilin-eosin technique, delaying closure of the wounds until the margins and surgical bed are tumor-free. RESULTS: We studied 83 patients with BCC; 49 were treated with the transoperatory technique and delay closure. We observed no recurrence in any patient that we followed up and there were no complications resulting from the technique in a 25-month follow up. CONCLUSIONS: We recommend this technique for tumors with high-risk of recurrence if Mohs' micrographic surgery is not available. PMID- 15125511 TI - Therapeutic potential of azithromycin in rosacea. AB - BACKGROUND: Systemic antibiotics currently used in the treatment of rosacea are sometimes associated with uncomfortable side-effects. Therefore, a need for an effective agent with few side-effects and good patient compliance exists. Azithromycin, a macrolide antibiotic with prolonged mode of action, has recently been found to be an effective alternative in the treatment of inflammatory acne. We planned a study to evaluate the efficacy and safety of azithromycin in rosacea. METHODS: An open-labeled study was performed in a population of 18 patients, with Plewig-Kligman stage 2 rosacea. Patients were given oral azithromycin for 12 weeks in decreasing doses. RESULTS: Fourteen subjects completed the trial. The treatment produced therapeutic benefits with regard to total scores as well as inflammatory lesion scores. At the end of 12 weeks, there was a 75% decrease in total scores (P < 0.001) and an 89% decrease in inflammatory lesion scores compared with basal values. Improvement continued during the 4 weeks after treatment. Adverse effects were minimal and well tolerated in most patients. CONCLUSION: Azithromycin is a promising agent in the treatment of rosacea with its few side-effects and good patient compliance. PMID- 15125512 TI - The importance of remembering Karel Fleischmann. PMID- 15125513 TI - Effect of myrrh extract on the liver of normal and bilharzially infected mice. An ultrastructural study. AB - In the present work, the efficacy of purified oloe-resin extract of myrrh derived from Commiphora molmol tree (commercially known as Mirazid) as a new, natural antischistosomal drug was investigated. The effect of myrrh on the ultrastructural profile of the non infected normal mice liver was also studied. Sixty male mice were used throughout this work and they were divided into 3 main groups (20 animals each): group I, non infected control animals, group II, infected animals and group III, infected animals treated with myrrh extract 8 weeks post infection (500 mg/kg body weight). The drug was given orally on an empty stomach after overnight fasting for five successive days. All animals were sacrificed after 12 weeks from the beginning of the experiment and small pieces of the liver were excised and prepared for ultrastructural study. The liver of the non infected animals which received myrrh extract (group IA) showed a more or less normal ultrastructural profile. Infected groups showed alterations of the ultrastructure of most of the hepatocytes with extensive intercellular fibrosis with abundant granulomas in the portal tract. In the infected treated group, most of the hepatocytes showed normal organelles with numerous microvilli extending into patent spaces of Disse. Marked reduction of granulomas in the portal areas and amelioration of intercellular fibrosis was also observed. On the basis of the observed results, it was concluded that myrrh extract has a promising antischistosomal non hepatotoxic activity. PMID- 15125514 TI - Seroepidemiological study of toxoplasmosis in rural areas in the eastern region of Saudi Arabia. AB - Serum from 1400 human subjects of different age, occupation, life style of both sexes was collected. Serological analysis for inactive toxoplasmosis (levels of IgG) and active toxoplasmosis (IgM) was done using the Microparticle Enzyme Immuno-Assay (MEIA) IMx-Toxo, IgG and IgM assays. In Al-Nereiyah and Al-Qurain, the overall seroprevalence of inactive toxoplasmosis was almost similar (26.36% and 25.0% respectively), while active toxoplasmosis was higher in Al-Qurain (4.91%) than in Al-Nereiyah (3.78%). The seroprevalence of IgG was similar in males and females in both areas, but increased with age and was higher in housewives, employees and farmers than in students and children. Almost a similar result was recorded for IgM in respect to age and occupation except that seroprevalence was higher in females than in males in Al-Qurain area. PMID- 15125515 TI - Detection of circulating antigens in sera from visceral leishmaniasis patients using dot-ELISA. AB - The anti-amastigote IgG antibodies were raised in rabbits purified and fractionated using the standard methods. From the same anti-amastigote IgG antibodies, a peroxidase conjugate reagent was prepared. Initially, the anti amastigote IgG antibodies and their corresponding peroxidase conjugate reagents were analyzed with respect to their ability to recognise the homologous antigens. Thirty five serum samples from visceral leishmaniasis patients (VLP) were examined using dot-ELISA. In addition, thirty seven serum samples from either non infected individuals or from patients infected with heterologous organisms were used to define the sensitivity and specificity of the test. Circulating antigens in 35 serum samples from VLP were successfully identified using dot-ELISA giving 100% sensitivity. With exception of serum samples from patients with Plasmodium vivax which shown cross-reactivity with Leishmania donovani antigens, the test appeared to be specific for the detection of circulating leishmanial antigens. Cross-reactivity was not observed with P. faciparum or Brucellosis. PMID- 15125516 TI - Merogony of Toxoplasma gondii (Apicomplexa: Coccidia) and its effect on the mortality and histopathology in the house mouse Mus musculus. AB - Merogonic stages of Toxoplasma gondii, their effects on the mortality and histopathological changes in the orally-inoculated male house mice Mus musculus with low and high doses (LD&HD) of T. gondii sporulated oocysts was investigated. The mortality percentage in LD group was 16%, 20%, 28% and 72%, while that in HD group was 36%, 56%, 72% and 100% in the 1st-4th week, respectively. Meanwhile, the maximum mortality percentage of the control group reached 6%. The histopathological changes induced by the parasite in different organs such as kidneys, liver, brain, spleen, heart, lungs and testes were discussed. PMID- 15125517 TI - Diagnostic microarray: an array of possibilities for parasitology. AB - Parasites affect a majority of the world's population. Despite this fact, dreams of developing vaccines remain far off. Scientists have long studied gene expression as a hallmark of gene activities reflecting current cell conditions. Analyzing differentially expressed genes is a major initiative, and most labs recoil at the amount of time and high costs required obtaining results. By employing microarrays, researchers can decrease their reliance upon time consuming techniques; consequently, microarray is beginning to dominate other molecular diagnostic technologies. Moreover, the ability of microarrays to monitor simultaneous gene expression of thousands of genes and to produce broad arrays of data has the potential to shift the resources of the scientists from data gathering to analyzing data that are already available. As microarray technology improves and its cost decreases, the role of ability to "see" the molecular biology pathways involved in parasite host relationships will place this technology at the forefront of parasite research. PMID- 15125518 TI - Molecular characterization of Egyptian human and anima Echinococcus granulosus isolates by RAPD-PCR technique. AB - Five primers of known, but arbitrary nucleotide sequence (OPH-03, OPH-05, OPH-12, OPH-15, OPH-18) were used to detect genetic variability in Egyptian human, camel and pig E. granulosus isolates. OPH-03, OPH-05 & OPH-15 proved useful as genetic markers of strain variation, while OPH-12 and OPH-18 allowed distinction at the genus level i.e. diversified from Cysticercus tenuicollis. OPH-03 was the most effective giving sharp distinct banding pattern and the least values of similarity coefficients. Some variations were detected within E. granulosus isolates from the same host. The level of heterogeneity was low in three of the human isolates, camel and pig strains. Individual variation was detectable within other 3 human isolates. Human and camel isolates were the most related pair, having similar patterns and the highest similarity coefficients. The study implies that human cases in Egypt are of the camel/dog strain, and camels are important hosts for the transmission of human hydatidosis. PMID- 15125519 TI - Evaluation of Capillaria philippinensis coproantigen in the diagnosis of infection. AB - In order to overcome the false negative diagnosis of infection with C. philippinesis at time of absence of eggs in stool, coproantigen prepared from stools of infected patients was evaluated serologically. This antigen was able to detect anti-Capillaria antibodies in the sera of infected cases at the same OD level produced with Capillaria crude worm antigen using indirect ELISA technique C. philippenensis coproantigen did not cross-react with sera from patients with schistosomiasis mansoni, fascioliasis or strongyloidiasis at 1:00 serum dilution. Laboratory-prepared hyperimmune sera versus crude worm antigen of C. philippinensis succeeded in capturing Capillaria antigen prepared from the stools of infected patients and did not cross react with coproantigens prepared from stool samples of cases infected with S. mansoni or Fasciola using sandwich ELISA technique. PMID- 15125520 TI - Treatment of human Demodex folliculorum by camphor oil and metronidazole. AB - A total of 15 females suffering from erythematotelangiectatic rosacea and 12 females free from other dermatological lesions were selected. Demodex folliculorum infestation density in both patients and control were evaluated by non-invasive skin surface biopsies. Five facial sites were selected. The daily topical application of 1/3 diluted camphor oil with glycerol and 500 mg metronidazole orally were given for fifteen days. The results were very successful with no clinical side effects. PMID- 15125521 TI - Microhabitat and surface ultrastructure of Neothoracocotyle commersoni and Pricea multae (Monogenea: Microcotyloidea) from gills of Scombrid fish, Scomberomorus commerson. AB - The microhabitat and surface topography of microcotylid monogenean parasites Neothoracocotyle commersoni and Pricea multae from gills of Spanish mackerels, Scomberomorus commerson was investigated. The results showed that N. commersoni was aggregated at the first gill sector while P. multae was aggregated at both first and second sectors in single infestation. In contrary, in mixed infestation, N. commersoni occupied the first sector of gills and P. multae was restricted only at the fourth sector with high intensity of infestation compared to N. commersoni. Aggregation of single species of monogenea in restricted habitat may aid cross-fertilization and hence producing of hybrids strains of parasites which might be more potent and vicious causing severe damage to the host. Surface ultrastructure of the representative monogenea showed that the tegument was provided with microvillus-like projections that suggested metabolic exchange and absorption of micromolecular nutrients from the surrounding environment. The architecture of clamps of N. commersoni was differed from that of P. multae in possessing microvillous-like projections which may act as organ of nutrition beside its principle attachment function. Ultrastructure of the clamps of both species described aid in understanding of the parasite habitat and survival. PMID- 15125522 TI - Seasonal distribution of Culicini larvae in Greater Cairo. AB - Five species of mosquito larvae were encountered in Greater Cairo, Culex pipiens, Cx. pusillus, Cx. perexiguus, Aedes caspius and Culiseta longiareolata. In parts of Qualyoubia G. Cx. pipiens was the most dominant and the least was Cx. perexiguus. In parts of Giza G. Cx. pipiens was the most dominant and the least was Cs. longiareolata. In Cairo G. Cx. pipiens was the most dominant and the least was Ae. caspius. The overall abundance in a descending order was Cx. pipiens (61.74%), Cs. longiareolata (15.56%), Ae. caspius (15.3%), Cx. pusillus (4.0%) and Cx. perexiguus (3.16%). However, in all sites Cx. pipiens were more in December than in August. The difference in number of Cx. pipiens in one hand and all number of larvae collected on the other hand was non significant. PMID- 15125523 TI - Myiasis in sheep farms in Riyadh Region, Saudi Arabia. AB - During a survey for myiasis in Riyadh Region, 3712 sheep were examined. 73 animals (2%) were infested with different dipterous larvae. Out of the 115 larvae recovered, 100 larvae (87%) were Chrysomya bezziana, 10 (8.7%) were C. albiceps, and 5 (4.3%) were Wohlfahrtia nuba. The prevalence rate of larval myiasis among young sheep was 60%, and 40% among adults. The myiasis incidences were highest during Mar-May (60%) and Sept-Nov (31.5%) where temperature and R.H. are optimum. In the dry hot season (Jun-August) and cold season (Dec-Feb) infestation incidences were low (5%&1.5% respectively). Temperature and R.H. are conducive in controlling myiasis in other regions of the Kingdom. PMID- 15125524 TI - Influence of temperature and salinity on the viability and infectivity of Giardia lamblia and Cryptosporidia parvum. AB - The present work was designed to assess the effect of different degrees of temperature and salinity on the viability and infectivity of G. lamblia and C. parvum at different storage time. The results revealed that boiling of protozoa for one minute minimized their viability to less than 1% and rendered them non infectious, while exposure to 4 degrees C and -4 degrees C up to seven days preserved their viability and infectivity. Whereas it was found that salinity was effective at high concentration (50 ppt) or for long storage time at lower concentrations. PMID- 15125525 TI - A simple micro-assay method for estimating blood meal size of the sand fly, Phlebotomus langeroni (Diptera: Psychodidae). AB - The accurate measurement of blood meal size in Phlebotomus langeroni, the potential vector of infantile visceral leishmaniasis in Egypt, is important to determine the number of parasites taken in fully engorged insects. A simple protein content micro-assay is introduced for that purpose. The accuracy of this method was confirmed by hemoglobin estimation method. Laboratory bred P. langeroni were fed artificially on defibrinated human blood and the fully engorged flies were carefully dissected on ice, within 1-10 min after feeding, since the time of dissection is critical. Serial concentrations of the defibrinated human blood were required as standards. Results show that the full blood meal taken by P. langeroni ranged from 0.76-0.94 mm3 of blood with a mean volume of 0.85 +/- 0.02 mm3 and from 0.71- 0.99 mm3 of blood with a mean volume of 0.83 +/- 0.02 mm3 as measured by protein content and hemoglobin estimation methods respectively. The data showed that there is no significant difference (P=0.27) between the two methods in estimating the blood meal size of P. langeroni. In addition, protein content micro-assay had the advantages of being accurate, rapid, sensitive and reliable. PMID- 15125526 TI - Association of some HLA-DRB1 antigens with Echinococcus granulosus specific humoral immune response. AB - An ELISA system using crude camel hydatid fluid antigen was used to detect specific IgG and IgG1 in sera of 35 cystic echinococcosis (CE) patients in whom the distribution of class II HLA-DR3 and HLA-DR11 was determined. The recorded sensitivities were 88.6 % and 94.35 % for IgG and IgG1 respectively. In patients with high humoral immune response a statistically highly significant increased frequency of HLA-DR3 (P<0.01) was recorded for IgG with high relative risk value (RR=3.2) and reasonable etiologic fraction (EF=0.3), while HLA-DR11 recorded P<0.05, RR=2.6, & EF=0.2. For IgG1 both antigens showed significant increased frequency (P<0.05), RR (2.95 & 2.79 respectively), EF (0.28 & 0.23 respectively). HLA-DR3 was highly significantly associated with complicated cases (P<0.01, RR=4.36, EF=0.4) in whom the mean antibody units for both IgG and IgG1 were significantly raised (P<0.05). It is advisable to rely on IgG1 for the diagnosis of CE and to consider the genetic disposition of the patient as an important criterion in the outcome of infection. PMID- 15125527 TI - Pulmonary trichomoniasis: improved diagnosis by using polymerase chain reaction targeting Trichomonas tenax 18S rRNA gene in sputum specimens. AB - A polymerase chain reaction (PCR) assay targeting species-specific region in 18 small subunit ribosomal RNA gene of Trichomonas tenax was used to examine sputum specimens in order to diagnose pulmonary trichomoniasis caused by T. tenax. It was compared with wet mount preparation, Giemsa-stained smear, and Kupferberg Trichononas broth culture for detection of T. tenax trophozoites in sputum. The study included 250 individuals; 100 immunocompromised patients with chest complaints (group I) and 100 patients with chronic pulmonary diseases (group II), and 50 healthy individuals as controls (group III). 20 cases among all examined were positive in one or more method giving for pulmonary trichomniasis a total prevalence of 8%; 12 cases (12%) in group I, 8 cases (8%) in group II, and none in group III, with no significant difference between groups I & II. Pulmonary trichomoniasis was prevalent at age ranged between 31 to 50 years, and in total males (10%) than females (5.5%) with no significant difference. Among the 200 examined patients, pulmonary trichomoniasis had a prevalence of 3% by wet mount, 2.5% by Giemsa-stained smear, 7% by culture, compared to 10% by PCR. Culture was used as reference standard. All culture positive specimens were PCR positive showing a product at 0.8 Kb long by agarose gel electrophoresis, and giving a 100% sensitivity. Wet mount, Giemsa-stained smear, and culture had a sensitivity of 43%, 35.7%, and 70%, respectively. No PCR negative specimens were positive by any of the other methods. 6 specimens were culture negative PCR positive and remained PCR positive when retested 3 times. The calculated specificity of PCR was 97%. NO PCR target product was amplified with DNAs of T. vaginalis and various pulmonary pathogens. The results are discussed. PMID- 15125528 TI - Molecular characterization of Egyptian Giardia lamblia isolates. AB - Six Giardia lamblia strains (4 from Egypt, one from the USA and one from Sudan) were used to study the phenotypic and genotypic variation in some Egyptian G. lamblia strains compared to other G. lamblia strains, which may be responsible for the difference in their behavioral characteristics. By using SDS-PAGE for antigenic study, E1 strain appeared different from the rest of the strains with two bands; one at 121 Kda and the other at 34 Kda which were not present in other strains, while bands at 130 and 43 Kda were present in all strains but absent in E1 strain. Another difference between E1 strain and the rest of the strains was obtained by CAE, using PGM enzyme where E1 strain gave a different zymodeme than the other strains. Based on the computerized RAPD- PCR analysis, 4 rapdemes were identified; rapdeme 1 contained E1 strain, rapdeme 2 contained E2, E3 and E4 isolates, rapdeme 3 contained Sudan strain and rapdeme 4 contained USA strain. In conclusion this study revealed diversity between G. lamblia strains especially E1 strain which showed unique characters. PMID- 15125529 TI - Toxicological efficacy of some indigenous dill compounds against the flesh fly, Parasarcophaga dux Thomson. AB - The effect of the LC50 of the three isolated compounds (apiol, myristicin and d carvone) from dill, Anethum graveolus on growth and reproduction of Parasarcophaga dux showed that three compounds especially apiol caused significant reduction in the percentage of adults emergence and females fecundity. The temperature toxicity relation shape of the three compounds and five insect growth regulations (methoprene, hydroprene, teflubenzuron, chlorfluazuron and Precocene I) alone or in combination against P. dux was studied and discussed. PMID- 15125530 TI - The influence of temperature and feeding on development, adult longevity and larviposition of Parasarcophaga hirtipes Wiedemann (Diptera: Sarcophagidae). AB - Number of sarcophagid flies was collected from the area of Dokki, Giza and kept in laboratory under normal weather conditions. Adult development, longevity and reproduction of Parasarcophaga hirtipes were studied in relation to variations of temperature and relative humidity during the period from January to May 2002. The mean longevity of adults of both sexes decreased with increase of temperature and decrease of relative humidity. The mean larval duration periods also significantly decreased with increased temperature but the mean pupae duration for both sexes did not change significantly during the experiment. The total development time from larvae to adults of both sexes decreased with increase of temperature. The mean longevity of adults female and male was affected by the presence of both sexes together every day rather than their presence together for only 3 days period to allow for one mating to occur. The effects of meat, liver and fish as food constituent on developmental period, longevity and larviposition of the fly were investigated under variation of relative humidity and temperature. The mean longevity of adult of both sexes was increased on feeding on meat. The change was not affected when fed on liver and fish. The mean development period of larvae and pupae and the total development period of both sexes not significantly increased with liver feeding. PMID- 15125531 TI - Enzyme activity in the flesh fly Parasarcophaga dux Thomson influenced by dill compounds, myristicin and apiol. AB - The newly emerged adults of Parasarcophaga dux were treated topically with various doses of myristicin and apiol isolated from roots of dill plant, Anethum graveolus. The compounds toxicity and the dehydrogenase activities of the treated stage and its subsequent developmental stages were studied. The results indicated that apiol more toxic than myristicin. The spectrophotometric evaluation exhibited changes in dehydrogenase activities after treatments. Compounds increased the activities of both alpha-GPDH and MDH in first half of metamorphosis (immature stages). But, the level of ME activities of the various stages was obviously decreased. PMID- 15125532 TI - Egyptian genotyping of Giardia lamblia. AB - Out of 105 patients infected with Giardia, 38 patients have Genotype I (36.19%), 13 have Genotype II (12.38%), 10 have Genotype III (9.52%), 16 have mixed Genotype infection (15.24%) and 28 with undetermined Giardia infection by PCR (26.67%). None of the control group gave positive results for Giardia in stool by PCR. So, the sensitivity of the test for detection and identification of Giardia Genotypes from the original stool samples was 73.33% and specificity was 100%. Out of 61 cases in the symptomatic group, the prevalence of Giardia Genotype I was 32.79%, Genotype II was 16.39%, Genotype III was 9.84%, mixed Genotype infection was 16.39% and undetermined Genotype was 24.59% as compared to 40.91%, 6.82%, 9.09%, 13.64% & 29.55% in the asymptomatic group respectively. There is statistically insignificant difference between both groups as regarding the prevalence of the different Giardia Genotypes. (P < or = 0.05). The use of PCR as a routine work for diagnosis of giardiasis is not accepted at least in the developing and under-developing countries due to its high cost, the high quality of technical staff and advanced laboratory equipments required for PCR performance. Its application is usually limited to research activities, the detection of water sources contamination and for the detection of a potential source of Giardia infection in epidemics. PMID- 15125533 TI - Identification of immunoreactive proteins of Trichinella spiralis adult and adult excretory/secretory (E/S) antigens in sera of human and animals. AB - Trichinella spiralis adult and adult excretory/secretory antigens were evaluated by ELISA and Western blot techniques for the serological diagnosis of trichinellosis in naturally infected human, swine and experimentally infected rats. Blood samples were collected from 16 symptomatic patients at hospital and 12 asymptomatic individuals working at the swine slaughter-house. Blood samples were also obtained from 75 pigs at two abattoirs and experimentally infected rats (day 25 PI). There was no great difference in the seroprevalence of trichinellosis between symptomatic (56%) and asymptomatic (50%) individuals. ELISA results recorded that 13% of swine was seropositive, while 44% were suspected to be infected with T. spiralis. Immunoblotting profiles of T. spiralis adult antigen against human, swine and rat sera showed common reactive bands at 95.00 and 64.466 KDa (human and swine), and 35.554 KDa (human and rat), while the blotting patterns of adult E/S products against human, swine and rat antibodies recognized two trichinellosis-specific determinants between human and swine (87.619 & 74.136 KDa) and between human and rat (98.00 & 16.535 KDa). It can be suggested that a 45.00, 75.355 & 25.389, and a 57.989 KDa polypeptides of adult antigen were diagnostic for human, swine and rat trichinellosis, respectively. While a 26.00, 24.00, and 46.994 KDa proteins of adult E/S products were diagnostic for human, swine and rat trichinellosis, respectively. PMID- 15125534 TI - The effect of severe cystic hydatidosis on the liver of a Najdi sheep with special reference to the cyst histology and histochemistry. AB - A single case of severe hydatidosis in a liver of a Najdi sheep was studied. The estimated age of animal was 1.5 years and the liver was infected with 29 cysts and their diameters ranged from 4.0 mm to 5.1 cm. Cysts with diameter less than 10 mm were immature and those more than 1.5 cm were fertile and developed protroscoleces. Harvested protroscolices measured 120x60 to 140x80 um. Comparative histochemical studies by haematoxylin & eosin, periodic acid Schiff (PAS) and Mossaon's trichrome stains revealed that the cystic wall consisted of three layers which are germinal, laminated and fibrous respectively. The thickness varied with each stain. The glycogen and mucopolysaccharide content increased in infected sheep. Histopathological changes were the formation of fibrotic capsules around biliary tracts and portal vein and also leaky liver was marked in all the sections examined. Meanwhile, pre-malignant changes were seen in the different foci particularly around the biliary tracts and portal veins. The histochemical structure of the hydatid cyst wall may pave the way for effective therapeutic treatment. PMID- 15125535 TI - Laboratory and field studies on oribatid mites as intermediate host of Moniezia expansa infecting Egytptian sheep. AB - Three species of oribatid mites, Scheloribates zaherii, Zygoribatula tadrosi and Z. sayedi from pure colonies were experimentally exposed to infection by allowing them to feed on stool sheep infected with Moniezia expansa. The mites were followed up to the development of the infective cysticercoids. M. expansa was able to achieve sucessfully its larval development in the three species of oribatid mites under laboratory conditions. These were demonstrated after 84, 73 & 69 days post infection, respectively. Z. tadrosi is recorded as inter-mediate host for the first time in Egypt. Six species of oribatid mites, Oppiella nova, S. laevigatus, S. zaherii, Xylobates souchiensis, Epilohmannia pallida aegyptiaca and Z. sayedi, recovered from the sheep infested farm soil, were found naturally infected with different developmental stages of M. expansa. PMID- 15125536 TI - Study of some immune aspects in patients with fascioliasis before and after Chommiphora molmol (Mirazid) treatment. AB - This study was planned to evaluate the in vitro production of IL-1 beta and IL-4 by peripheral blood mononuclear cells (PBMCs) and total IgE in patients with fascioliasis before and 3 months after treatment with purified extract of myrrh from Commiphora molmol tree (Mirazid), to determine the role of these variables in immunopathogenesis of the disease in relation to this new drug. The study was carried out in Departments of Tropical Medicine, Al-Azhar University Hospitals in the period from March 2002 to November 2003. A total of 35 patients with chronic fascioliasis with age range from 9-45 years in addition to 10 healthy subjects with matched age and sex serving as controls were studied. Serum IgE and in vitro IL-1 and IL-4 were estimated by enzyme immuno-assay (ELISA) before and 3 months after therapy. Results revealed significant increase in IL-1 beta in patients before treatment than control (p<0.001) but it decreased significantly after therapy (p<0.001) to reach the control level (p=0.16). In contrast, IL-4 was significantly lower than control before therapy (p=0.04) and increased significantly after treatment (p<0.001) to reach normal levels as control (p=0.59). Total IgE was significantly elevated in patients before treatment (p<0.001) and it did decrease significantly with treatment (p<0.001), although it remained significantly higher than the control level. In conclusion, Mirazid is an effective fasciolicidal drug. IL-1 may be involved in disease immunopathogenesis and the depressed IL-4 may be a phenomenon of parasite immune suppression. Complete decline of total IgE is not an early criterion of cure. PMID- 15125537 TI - Adaptation of three Babesia divergens isolates to continuous culture in rat erythrocytes. AB - Three Babesia divergens isolates have been cultured continuously for 6 months in rat erythrocytes using the candle jar technique. One isolate was already rat adapted, the other two became adapted to rats through continuous culturing in rat erythrocytes. Babesia was cultured in rat erythrocytes in RPMI medium supplemented with 20% foetal calf serum. The highest parasitaemia was 35% and multi-parasitization of red blood cells was often observed. Cultures of B. divergens remained infective to splenectomized rats. Cultures with high parasitaemias contained a large number of extra-cellular merozoites, when separated from the red blood cells, they retained their infectivity. PMID- 15125538 TI - Science, medicine and society: biomedical ethics of the new millennium. PMID- 15125539 TI - Coinfection with Listeria monocytogenes potentlxtes the response of BALB/c mice against Leishmania major. AB - Protection against L. major is dependent on the stimulation of an anti leishmanial T helper1 (Th1) response and the production of Interferon (IFN)-y. BALB/c mice develop a Th2 response and fatal infection with Leishmania major. Strategies that boost IL-12 production have shown to be protective. The innate response to Listeria monocytogenes is associated with IL-12 production. The co infection of BALB/c mice with L. monocytogenes attenuates the course of L. major infection. Lesion sizes were smaller, and co-infected mice out-survived controls injected with L. major alone. The parasite load was reduced at site of injection, in draining lymph nodes (LN) and spleen. During the first week of infection, in vitro Leishmania-restimulated LN cells from co-infected mice produced higher levels of IFN-7 and undetectable levels of IL-4 compared to controls. Significant IL-4 mRNA expression was detected in LN cells of control but not in co-infected mice. PMID- 15125540 TI - Assessment and management of air quality for an opencast coal mining area. AB - A study for the assessment and management of the air quality was carried out at Lakhanpur area of Ib Valley Coalfield in Orissa state in India. The 24-h average concentrations of total suspended particulate (TSP) matter, respirable particulate matter (PM10), sulphur dioxide (SO2) and oxides of nitrogen (NOx) were monitored during 1 year period. Samplings were done at a regular interval through out the year at 13 monitoring stations in the residential areas and four monitoring stations in the mining/industrial areas. The 24-hr average TSP and PM10 concentrations ranged from 338.8 to 799.8 microg m(-3) and 102.5-425.6 microg m(-3) for industrial area, and 72.3-497.1 microg m(-3) and 40.8-171.9 microg m(-3) for residential area, respectively. During the study period 24-hr and annual average TSP and PM10 concentrations exceeded the respective standards set in the national ambient air quality standard (NAAQS) protocol at most residential and industrial areas. However, 24-hr and annual average concentrations of SO2 and NOx were well within the prescribed limit of NAAQS both in the residential and industrial areas. The annual and 24-hr average concentrations varied from 23.3 to 36.8 microg m(-3) and 16.0-55.2 microg m(-3) for SO2 and 23.9-41.9 microg m(-3) and 19.0-58.1 microg m(-3) for NOx, respectively. The temporal variations of TSP and PM10 fitted polynomial trend with an average correlation coefficient (R2) of 0.77 (+/-0.17) for TSP and 0.85 (+/-0.10) for PM10. On average the PM10 in the ambient air of the mining area constituted 31.94 (+/-1.76)% of the TSP. The linear regression correlation coefficient (R2) between TSP with PM10 and NOx with SO2 was 0.86 (+/-0.12) and 0.57 (+/-0.20), respectively. Maximal concentrations of TSP and PM10 occurred within the mining site based on the kriging technique. A management strategy is formulated for effective control of air pollution at source and other mitigative measures recommended including implementation of green belts around the sensitive areas where the concentration of air pollutants exceeded the standard limit. PMID- 15125541 TI - Analysis of the prescribed burning practice in the pine forest of northwestern Portugal. AB - The ignition of low-intensity fires in the dormant season in the pine stands of north-western Portugal seeks to reduce the existing fuel hazard without compromising site quality. The purpose of this study is to characterise this practice and assess its effectiveness, based on information resulting from the normal monitoring process at the management level, and using operational guidelines, fire behaviour models and a newly developed method to classify prescribed fire severity. Although the region's humid climate strongly constrains the activity of prescribed fire, 87% of the fires analysed were undertaken under acceptable meteorological and fuel moisture conditions. In fact, most operations achieved satisfactory results. On average, prescribed fire reduces by 96% the potential intensity of a wildfire occurring under extreme weather conditions, but 36% of the treated sites would still require heavy fire fighting resources to suppress such fire, and 17% would still carry it in the tree canopy. Only 10% of the prescribed burns have an excessive impact on trees or the forest floor, while 89% (normal fire weather) or 59% (extreme fire weather) comply with both ecological integrity maintenance and wildfire protection needs. Improved planning and monitoring procedures are recommended in order to overcome the current deficiencies. PMID- 15125542 TI - Emissions study of co-firing waste carpet in a rotary kiln. AB - Post-consumer carpet represents a high volume, high energy content waste stream. As a fuel for co-firing in cement kilns, waste carpet, like waste tires, has potential advantages. Technological challenges to be addressed include assessing potential emissions, in particular NO emissions (from nylon fiber carpets), and optimizing the carpet feed system. This paper addresses the former. Results of pilot-scale rotary kiln experiments demonstrate the potential for using post consumer waste carpet as a fuel in cement kilns. Continuous feeding of shredded carpet fiber and ground carpet backing, at rates of up to 30% of total energy input, resulted in combustion without transient puffs and with almost no increase in CO and other products of incomplete combustion as compared to kiln firing natural gas only. NO emissions increased with carpet waste co-firing due to the nitrogen content of nylon fiber. In these experiments with shredded fiber and finely ground backing, carpet nitrogen conversion to NO ranged from 3 to 8%. Conversion increased with enhanced mixing of the carpet material and air during combustion. Carpet preparation and feeding method are controlling factors in fuel N conversion. PMID- 15125543 TI - The application of bioflocculant for the removal of humic acids from stabilized landfill leachates. AB - The evaluation of bioflocculant, in comparison with traditional inorganic coagulants, for the removal of humic acids from landfill leachates stabilized by biological treatment, was performed using conventional jar-test coagulation experiments. The optimized conditions (pH and coagulant dosage) were identified for the treatment of synthetic solutions as well as for biologically pre-treated landfill leachates. It was found that the application of bioflocculant was quite efficient in the removal of humic acids from synthetic solutions as well as in the reduction of COD content from real landfill leachates. The optimal pH value was found to be between 7 and 7.5, while a 20 mg/l bioflocculant dosage was sufficient in providing more than 85% humic acid removal. The results were comparable with those obtained by the application of conventional coagulants such as alum or polyaluminum chloride; therefore, bioflocculant can be considered as a viable alternative in the treatment of landfill leachates applying coagulation. PMID- 15125544 TI - Photocatalytic degradation of organic wastes by electrochemically assisted TiO2 photocatalytic system. AB - Photocatalytic degradation of organic wastes with nanosized titanium dioxide particles has been studied for a long time in order to offer an appropriate method for wastewater treatment, but its practical application is greatly limited by the slow process. In this work, an electrochemically assisted TiO2 photocatalytic system was set-up by combining a TiO2 photocatalytic cell with a three-electrode potentiostatic unit. The composite system revealed high photocatalytic activity towards organic wastes mineralization. After continuous treatment for 0.5 h, the maximum absorption of rhodamine 6G (R-6G) was reduced by more than 90%; chemical oxygen demand (COD) and biochemical oxygen demand (BOD5) of textile dye wastewater (TDW) were decreased by 93.9 and 88.7%, respectively. The biodegradability of TDW was also improved because the COD/BOD5 ratio decreased from 2.1 to 1.2. All these results indicated that the composite system could be used for effective organic wastes mineralization or as a feasible detoxification and color removal pretreatment stage for biological post treatment. PMID- 15125545 TI - Quantifying the human health benefits of curbing air pollution in Shanghai. AB - Urban development in the mega-cities of Asia has caused detrimental effects on the human health of its inhabitants through air pollution. However, averting these health damages by investing in clean energy and industrial technologies and measures can be expensive. Many cities do not have the capital to make such investments or may prefer to invest that capital elsewhere. In this article, we examine the city of Shanghai, China, and perform an illustrative cost/benefit analysis of air pollution control. Between 1995 and 2020 we expect that Shanghai will continue to grow rapidly. Increased demands for energy will cause increased use of fossil fuels and increased emissions of air pollutants. In this work, we examine emissions of particles smaller than 10 microm in diameter (PM10), which have been associated with inhalation health effects. We hypothesize the establishment of a new technology strategy for coal-fired power generation after 2010 and a new industrial coal-use policy. The health benefits of pollution reduction are compared with the investment costs for the new strategies. The study shows that the benefit-to-cost ratio is in the range of 1-5 for the power sector initiative and 2-15 for the industrial-sector initiative. Thus, there appear to be considerable net benefits for these strategies, which could be very large depending on the valuation of health effects in China today and in the future. This study therefore provides economic grounds for supporting investments in air pollution control in developing cities like Shanghai. PMID- 15125546 TI - Some observations on the terminology in co-operative environmental management. AB - The notion of managing resources in partnership flourishes in natural resources literature. The terms partnership, collaboration, and co-management are associated with co-operative environmental management. Examining issues of definition reveals similarities, differences and, at times, imprecise use of the three terms. The potential for clarity prompts the proposal of a multi dimensional model of co-operative environmental management. The model consists of three key dimensions. The first dimension reflects the extent power is shared among the actors and agencies involved in the agreement. The second dimension delineates who is involved in the management regime. Process, the final dimension, reflects the variety of ways in which co-management may function or proceed. The presented model highlights dimensions requiring attention by those working within co-operative environmental management. The model is valuable as it reflects the complexity and range of such arrangements in practice. PMID- 15125547 TI - Spatial patterns of recreation impact on experimental campsites. AB - Management of camping impacts in protected areas worldwide is limited by inadequate understanding of spatial patterns of impact and attention to spatial management strategies. Spatial patterns of campsite impact were studied in two subalpine plant communities in the Wind River Mountains, Wyoming, USA (a forest and a meadow). Response to chronic disturbance and recovery from acute disturbance were both assessed. Previously undisturbed sites were camped on at intensities of one and four nights/year, for either one or three consecutive years. Recovery was followed for three years on sites camped on for one year. Percent bare ground, assessed in 49 contiguous 1 m2 quadrats, increased with increasing use frequency, particularly on the forest sites. Magnitude of impact varied spatially within campsites, with impact decreasing as distance from the center of the campsite increased. On the more fragile forest sites, this radial impact pattern developed rapidly and remained after three years of recovery. Concentration of camping activities around a centrally located small cooking stove was the apparent cause of this pattern. Maximum variation in magnitude of impact occurred at intermediate levels of campsite use and disturbance. The magnitude, variability and spatial pattern of impact varied with the spatial scale of analysis. Generally, results of these controlled experiments are consistent with earlier studies of campsites and validate the management implications derived from those studies. Even where campers use low-impact techniques, low levels of camping use can cause substantial impact and it is important to concentrate use. On resistant sites, however, it is possible that low levels of use can be sustained with minimal resultant impact. PMID- 15125548 TI - Application of the pressure-state-response framework to perceptions reporting of the state of the New Zealand environment. AB - The Pressure-State-Response framework for environmental reporting was used as a basis to develop a long-term study of people's perceptions of the state of the New Zealand environment. A postal survey of 2000 people, randomly drawn from the New Zealand electoral roll was used to gather data--an effective response rate of 48% was achieved. A range of different resource sectors was examined. We report on New Zealand's air, native animals and plants, and marine fisheries, as well as New Zealand compared to other developed countries. Respondents generally considered that in terms of pressures, states and responses, New Zealand was performing better than other developed countries and that for the resources examined here overall performance was in the adequate to good range, except for marine fisheries. The survey appears to be a useful tool for linking perceptions data into State of the Environment reporting. It also helps identify policy issues where perceptions do not match other scientific evidence or management initiatives. Such findings can be important for the successful implementation of policy measures. PMID- 15125549 TI - Promoting patient safety through the development of a pediatric chemotherapy and biotherapy provider program. PMID- 15125550 TI - Levels of total fungus and Aspergillus on a pediatric hematopoietic stem cell transplant unit. AB - The purpose of this descriptive study was to determine the levels of total fungus (TF) and Aspergillus in a pediatric hematopoietic stem cell transplant (HSCT) unit. One hundred twenty air samples and 120 floor samples were collected from the same locations in 10 patient rooms and bathrooms for 4 consecutive days. The count in colony-forming units of TF and Aspergillus from each of the samples was measured by the institution's mycology laboratory. Means, standard deviations, minimum values, and maximum values were determined for levels of TF and Aspergillus from different locations and on different days in the air and on the floor. Determination of a mean value of TF and Aspergillus for each room allowed for analysis of mean values of TF and Aspergillus for sample category, room side, room type, and room status. After visual examination of the mean values for the air samples collected, it was determined that the TF and Aspergillus in the air were less than the institution's acceptable air baseline standard. t tests and analysis of variance were used to verify the findings. PMID- 15125551 TI - Insuflon versus subcutaneous injection for cytokine administration in children and adolescents: a randomized crossover study. AB - Pain is a frequent complication of subcutaneous cytokine injections in children. A randomized crossover trial was conducted to determine the least painful and preferred method of cytokine administration for children and young people. The current standard practice of subcutaneous injection was compared with the use of Insuflon (Maersk Medical, Roskilde, Denmark), a subcutaneous indwelling catheter. Children aged between 1 month-and 18 years undergoing treatment within the oncology/hematology unit of a single tertiary hospital and receiving cytokines were eligible for the study. Twenty children participated in the study, each child receiving both administration methods in random order during sequential cytokine treatment courses. There was a trend toward higher pain scores when using subcutaneous injections for drug administration compared to Insuflon. Seventy-five percent (n = 15) of the children who completed the trial and their families preferred using insuflon for subcutaneous drug administration. Consideration needs to be given, however, to those who refused to enter the study, withdrew, or continued because of a preference for subcutaneous injections. Current practice at the Women's and Children's Hospital is to allow the child and parents to choose their preferred treatment modality for subcutaneous drug administration. PMID- 15125552 TI - Childhood brain tumors: parental concerns and stressors by phase of illness. AB - The objective of this study is to identify common problems and helpful resources important to parents of children with brain tumors by illness phase and to determine associations with stress. Parents with a child diagnosed within the past 10 years were surveyed regarding healthcare provider interactions, medical information/education, health care utilization and psychosocial concerns. Survey items were rated as problems or helpful, and for importance at each phase of illness. Stress was recorded from 0 to 10 for each phase; associations with demographic characteristics and items were tested statistically. A total of 139 parents from 87 families responded, with 45 mother-father pairs. Half reported unmet informational needs as most important during diagnosis (etiology), recurrence (complementary therapy), end of life (dying process), and remission (long-term effects). Mothers experienced greater stress than fathers during adjuvant treatment (p = .009). Stress increased (p < .05) during diagnosis and hospitalization/surgery with being married, at hospital discharge because of changes in child's personality/moods, during adjuvant treatment with unmet informational needs regarding stopping treatment, during recurrence regarding employment concerns, and during remission with unmet informational needs regarding life-time expectations. Stressors changed across phases of illness. Married respondents appeared at increased risk for stress. Further work is needed to tailor and evaluate interventions to decrease stress during illness phases. PMID- 15125553 TI - Childhood and adolescent cancer survivors' knowledge of their disease and effects of treatment. AB - Most children diagnosed with cancer will become survivors of their disease. However, a large number of these children may be at risk for the development of late complications. It is not clear whether these survivors and their families are aware of their treatment history including diagnosis, treatment, and the late complications of their treatment. The purpose of this needs assessment was to ascertain survivors' knowledge of their disease, the various treatment modalities, and whether they were aware of their individual risks for developing late complications of their treatment. A brief six-item questionnaire was developed to determine survivors' knowledge of their cancer treatment and to be completed just before at their survivor visit. A total of 141 survivors completed this questionnaire. The mean age at diagnosis was 6.8 years, and the median age at the time of this assessment was 16 years. Although all of the subjects stated they knew their disease, only 84% (n = 118) listed their diagnosis. The majority of the survivors knew they received chemotherapy, but only 50% were able to list one or more specific drugs they received. Further lack of knowledge was also evident for survivors who received radiotherapy. The results of this assessment represent a lack of knowledge especially of the survivors' individual risk for developing late complications of their therapy. Education about late complications of therapy should be introduced early and often such as at diagnosis, within months of the completion of therapy, and during every survivor clinic visit. A complete treatment summary should be provided to all survivors. This summary should include the survivors' individual risks for developing late complications and how their own health behaviors may influence the development of these late complications. PMID- 15125554 TI - Adventure therapy: a mental health promotion strategy in pediatric oncology. AB - In adventure therapy (AT), health professionals and adolescents with cancer come together to explore the wilderness of nature. One goal of this therapy is to encourage the adolescents to enhance their self-concept as part of an overall physical, cognitive, emotional or spiritual, social and psychological, or developmental rehabilitation that promotes health. The adolescents with cancer who participate in AT also learn about themselves through self-evaluation, self exploration, self-reevaluation, self-acceptance, and self-realization. Mental health promotion (MHP) is considered a perspective and a strategy to promote health. An AT experience could be an example of an MHP initiative in which nurses can take a leadership role in participating, and further investigating, the health effects of AT on adolescents with cancer. PMID- 15125556 TI - Reaching the underserved: lessons learnt from leprosy. PMID- 15125555 TI - A relic that disturbs. PMID- 15125557 TI - Progress of leprosy elimination in South-East Asia Region. PMID- 15125558 TI - Achievements of National Leprosy Eradication Programme (NLEP) in India. PMID- 15125559 TI - Cleaning of leprosy case registers in Chhattisgarh. AB - Cleaning of register exercise was based on examination of records and not of patients. Because of this exercise, names of 5676 patients were deleted for various reasons viz. completed FDT, Defaulter, AMDT etc. The findings of the present study are more or less similar to the finding of similar other studies carried out in different countries from time to time. Updating of leprosy registers should be a routine activity. For this sensitization of health functionaries at various level must be carried out about importance of cleaning of register and for systematic effort to review the registers at least on sample basis. PMID- 15125560 TI - Elimination of leprosy from India--prospects & challenges. PMID- 15125561 TI - Leprosy scenario in Goa. PMID- 15125562 TI - From vertical to integration for elimination: recent changes in the National Leprosy Eradication Programme in India. PMID- 15125563 TI - Status of leprosy in Uttar Pradesh. PMID- 15125564 TI - Leprosy in urban areas--a problem for elimination. PMID- 15125565 TI - [Pyrido [3,2-b]indol-4-yl-amines--synthesis and investigation of activity against malaria]. AB - Pyrido[3,2-b]indol-4-yl-amines--synthesis and investigation of activity against malaria Starting with 3-aminoindole-2-carboxylic acid ester 1 the annulated pyrido[3,2-b]indoles 6 and 8 were synthesized as key substances. The 4 chloropyridine derivative 8 reacted with the phenol Mannich bases 11 and the novaldiamine base 13, respectively, to yield the amodiaquine and cycloquine analogues 12 as well as the chloroquine analogue 14. The stability of the compounds 12 and 14 were proven by the half wave potentials measured by differential pulse voltammetry. Compounds 12 and 14 were tested for in vitro antimalarial activity using a chloroquine sensitive and a chloroquine resistant Plasmodium falciparum strain. The highest activity was shown by 12g with IC50 values of 50 nM and 38 nM, respectively. The in vivo activity of 12g was tested in Plasmodium vinckei infected mice resulting in ED50 values of 22 mg/kg and 26 mg/kg after intraperitoneal and oral administration, respectively. PMID- 15125566 TI - NMR regulatory analysis: determination and characterization of S-adenosyl-L methionine in dietary supplements. AB - 1H NMR methodology is described for the determination and characterization of the dietary supplement S-adenosyl-L-methionine (SAM), recently introduced to the US market, utilizing a 400 MHz spectrometer without the need of pure reference standards. The developed methodology is able to assess chemical structure, differentiate between biologically-active (S)-diastereomer and biologically inactive (R)-diastereomer, and determine the ratio of each in the dietry supplement formulation. The determination of the percentage of declared SAM was based on the integrals for the methyl proton of 2-methyl-2-propanol served as an internal standard at delta 1.24 and the methine proton H1' of SAM ribose ring at delta 6.06. The percentage of the active diastereomer was calculated from the relative intensities of the sulfonium methyl singlets corresponding to the major component (S)-diastereomer at delta 2.98 and the minor (R)-counterpart at delta 2.93. The accuracy was established by analyzing synthetic mixtures of the analyte and the internal standard. Excellent agreement was verified between the assay results and the quantities of analyte in the mixture. The mean +/- SD recovery values for SAM and its (R)-diastereomer impurity from a set of 10 synthetic mixtures were 99.6 +/- 0.8% and 22.5 +/- 0.1%, respectively. Using 10 lots, the percentage of SAM ranged from 0 to 110.7% of the declared value and the percentage of the active (S)-diastereomer ranged from 0 to 82.3%. PMID- 15125567 TI - Validation of a spectrophotometric method for quantification of xanthone in biodegradable nanoparticles. AB - Xanthone has been incorporated for the first time in nanoparticles of poly(D,L lactide-co-glycolide) (PLGA). For this purpose the estimation of xanthone content in the nanoparticles is a crucial tool for guaranteeing the reliability of the results. Thus, a simple spectrophotometric method was validated according to USP25 and ICH guidelines for its specificity, linearity, accuracy and precision. The method was found to be specific for xanthone in the presence of nanoparticle excipients. The calibration curve was linear over the concentration range of 0.5 to 4.0 microg/mL (r > 0.999). Recovery of xanthone from nanoparticles ranged from 86.5 to 95.9%. Repeatability (intra-assay precision) and intermediate precision were found to be acceptable with relative standard deviations values (RSD) ranging from 0.3 to 3.0% and from 1.4 to 3.1%, respectively. The method was found to be suitable for the evaluation of xanthone content in nanoparticles of PLGA. PMID- 15125568 TI - Capillary zone electrophoresis determination of loratadine in tablets. AB - A capillary electrophoretic method was developed for the determination of loratadine in pharmaceutical formulations. Capillary zone electrophoresis (CZE) separation and UV absorbance photometric detection were carried out in a 160 mm capillary tube with a 300 microm internal diameter, hydrodynamically (membrane) closed. The influences of pH, carrier cation and counter ion on the migration parameters of loratadine were studied and the following conditions were selected: 24 mmol/l glycine as a carrier cation, 1.6 mmol/l citric acid and 84 mmol/l acetic acid as counter ions at pH 3.2, 100 microA and 25 degrees C. The proposed electrophoretic method was successfully validated. It was convenient for the sensitive, simple, rapid and highly reproducible assay of loratadine. The determination of loratadine in tablet forms was demonstrated as an application of the method. CZE analysis was completed within 6 min. The detection limit of loratadine was 1.96 micro/mol/l at a 240 nm detection wavelength and the relative standard deviation for its determination was 0.6% for migration time and 1.1% for peak area. CZE in a hydrodynamically closed separation system, used for the first time for the analysis of loratadine, should also be convenient for complex biological sample applications, as it is easily combinable online with the purification CE modes (e.g. ITP). PMID- 15125569 TI - Melanoma vaccine based on the vector of membrane fusogenic liposomes. AB - Membrane fusogenic liposomes can deliver encapsulated contents into the cytoplasm directly in a Sendai virus fusion-dependent manner. Based on the high delivery rates into the cytoplasm, membrane fusogenic liposomes were investigated as an antigen delivery vehicle. The membrane fusogenic liposomes were formulated by fusing simple liposomes with ultraviolet inactivated Sendai virus. The vaccine was prepared by encapsulating mixture antigen into the simple liposomes, and then fusing with Sendai virus. The antigen, mixture proteins were extracted from B16 melanoma cells. Membrane fusogenic liposomes were characterized for their sizes and shape by laser light granule analysis instrument and transmission electron microscope (TEM). The cytotoxic T lymphocyte (CTL) responses level was evaluated by the 51Cr release method. The positive expression of CD4+ and CD8+ cells, entrapment capacity of membrane fusogenic liposomes and the concentration of IgG in serum of immunized mice were measured. The vaccine, formulated with simple liposomes can induced systemic responses, but not CTL responses. However, membrane fusogenic liposomes-formulated melanoma vaccine can elicit not only systemic immune responses but also strong CTL responses, and inhibit the accretion of tumor. Membrane fusogenic liposomesas a vector for use in anti-tumor vaccine therapy are feasible. PMID- 15125570 TI - Citric acid as a pH-regulating additive in granules and the tablet matrix in enteric-coated formulations for colon-specific drug delivery. AB - Colon-specific drug-delivery systems have been extensively investigated over the last decade. The aim of the study reported here was to investigate whether times of commencement of drug liberation and absorption could be controlled by varying the amount of citric acid in granule cores or in the tablet matrix in enteric coated multiple-unit tablets. One of the most important aims was to determine the optimal amounts and locations of citric acid in formulations intended as drugs targeted at the colon. Ibuprofen was used as the model drug. Drug release rates were studied in phosphate buffer at pH 6.8 and 7.4. A gradient dissolution study at pH 1.2, 6.8 and 7.4 was undertaken with two formulations. Drug absorption was studied by means of bioavailability tests. We concluded that the drug release rate could be controlled in vitro by changing the amount of citric acid in granule cores or the tablet matrix. In vivo tests confirmed that between 10 and 15% citric acid in the tablet matrix delayed the commencement of drug absorption most. This kind of formulations could be suitable for preparation of colon specific dosage forms. It is probably unnecessary to include citric acid in granule cores. No logical correlation between in vitro and in vivo results was obtained. PMID- 15125571 TI - Preparation and evaluation of microemulsion of vinpocetine for transdermal delivery. AB - Poorly soluble vinpocetine was selected as the model drug to prepare a microemulsion in order to increase solubility and in vitro transdermal delivery of the drug. Oleic acid was chosen as the oil phase due to its excellent solubilizing capacity. PEG-40 hydrogenated castor oil (Cremophor RH40) was employed as a surfactant (S) and purified diethylene glycol monoethyl ether (Transcutol P) was used as a cosurfactant (CoS). The effects of diverse types of oil, different weight ratios of surfactant to cosurfactant (S/CoS) on the solubility and permeation rate of vinpocetine were investigated. The optimized microemulsion consisted of 1% vinpocetine, 4% oleic acid, 20% Cremophor RH40, 10% Transcutol P and 65% distilled water (w/w), in which drug solubility was about 2,100 fold compared to that in water and the apparent permeation rate across the excised rat skin was 15.0 +/- 2.5 microg/cm2/h. Finally the physicochemical properties of the optimized microemulsion including pH, viscosity, refractive index, conductivity and particle size distribution were examined, which showed stable behavior after more than 12 months at ambient temperature. The irritation study showed that optimized microemulsion was a safe transdermal delivery system. PMID- 15125572 TI - Formation of particles in aqueous infusions of the medical plant Harungana madagascariensis. AB - In some aqueous plant extracts the formation of nanoparticles, microparticles and macroparticles has been observed. In the present investigation the particle formation in aqueous infusions of Harungana madagascariensis LAM. EX. POIR., a medicinal plant, was investigated using photon correlation spectroscopy (PCS) and scanning electron microscopy (SEM). The results show that nanoparticles with mean diameters of 220 nm are formed in aqueous infusions of the dried leaves of Harungana. The particles have an almost spherical shape. In aqueous infusions of the dried stem bark nanoparticles with a mean diameter of 242 nm (PCS) are observed at 25 degrees C. The particle size distribution has a maximum in the range of 200 nm to 300 nm. Nanoparticles can be detected in infusions of the leaves and the bark in the range of 25 degrees C to 55 C. The mean diameter of the nanoparticles in preparations of the bark is temperature dependent: At 55 degrees C the mean diameter is 144 nm, at 30 degrees C 197 nm and at 25 degrees C 242 nm. Lower temperatures result in higher count rates. In infusions of the leaves the mean diameters vary between 220 (25 degrees C) and 139 nm (55 degrees C). The particle formation was investigated at pH 2.2, 4.0 and 7.4 at 37 degrees C. Nanoparticles are detected in infusions of the leaves and the stem bark at each pH. The pH value has an influence on the mean diameter and the count rate. PMID- 15125573 TI - Limitation of Potts and Guy's model and a predictive algorithm for skin permeability including the effects of hydrogen-bond on diffusivity. AB - The Potts and Guy's model for skin permeability, log P = alpha log K - beta MV + delta where P is the permeability coefficient of a compound from aqueous solution through human skin in vitro, K and MV are octanol-water partition coefficient and molecular volume of the compound respectively, and alpha, beta, delta are constants, is examined for a data set of 53 miscellaneous compounds. The model will result in over-estimation for penetrants having higher hydrogen-bond donor activity and underestimation for penetrants having no hydrogen-bond donor. A predictive algorithm for skin permeability including the effects of hydrogen-bond on diffusivity is proposed: log P = alpha log K - beta MV - gamma Hb + delta where Hb is the descriptor of hydrogen-bonding capacity of penetrants and gamma is a constant. The calculated log P values from the latter model are in good accordance with respective experimental ones for the data set. PMID- 15125574 TI - Protective effects of luteolin-7-glucoside against liver injury caused by carbon tetrachloride in rats. AB - Ixeris chinensis (Thunb.) Nakai has been used as a Chinese folk medicine; the information on the physiological and biochemical functions of the compounds extracted from I. chinensis is still scanty. We investigated the effects of luteolin -7-glucoside (LUTG) isolated from I. chinensis against liver injury caused by carbon tetrachloride (CCl4). CCl4 significantly increased the enzyme activities of glutamic pyruvic transaminase (GPT) and glutamic oxaloacetic transaminase (GOT) in blood serum, as well as the level of malondialdehyde (MDA) and 8-hydroxydeoxyguanosine (8-OHdG) in liver tissue, and decreased the levels of reduced glutathione (GSH). Pretreatment with LUTG was not only able to suppress the elevation of GPT, GOT, MDA and 8-OHdG, and inhibit the reduction of GSH in a dose-dependent manner in vivo, but also reduce the damage of hepatocytes in vitro. On the other hand, we also found LUTG has strong antioxidant activity against reactive oxygen species (ROS) in vitro in a concentration-dependent manner. The hepatoprotective activity of LUTG was possibly due to its antioxidant properties, acting as scavengers of ROS. These results obtained in vivo and in vitro suggest that LUTG had protective effects against hepatic oxidative injury induced by chemicals. Further studies on the pharmaceutical functions and immunological responses of LUTG may help in the development of a clinical application. PMID- 15125575 TI - Cytotoxic activity of methanol extracts from Basidiomycete mushrooms on murine cancer cell lines. AB - Crude methanol extracts of 58 mushroom species were screened for their cytotoxic activities against two murine cancer cell lines, L1210 and 3LL, using the tetrazolium assay. A majority of extracts (74%) exhibited IC50 > 100 microg/ml against both cell lines. A most marked activity against one of the cell lines was noted for nine species (14% of the tested species). While Amanitales and Russulales tested were not found active, Polyporales and Boletales gave better results. Four species exhibited a significant cytotoxic activity (IC50 < or = 20 microg/ml) against at least one of the two murine cancer cell lines (Ganoderma lucidum, Meripilus giganteus, Suillus granulatus, S. luteus). The last one had never been investigated for its cytotoxic compounds before. PMID- 15125576 TI - Steroidal saponins from Dioscorea panthaica and their cytotoxic activity. AB - A new steroidal saponin, dioscoreside E (1), and a known compound, protodioscin (2), were isolated from an ethanol extract of the rhizomes of Dioscorea panthaica. The structure of 1 was established as 3-O-[bis-alpha-L-rhamnopyranosyl (1 --> 2 and 1 --> 4)-beta-D-glucopyranosyl]-26-O-beta-D-glucopyranosyl-20(R) methoxy-25(R)-furosta-5,22(23)-diene-3beta,26-diol, on the basis of spectral and chemical evidence. Compounds 1 and 2 showed cytotoxic activity against a panel of tumor cell lines. PMID- 15125577 TI - Immunosuppressive auronol glycosides from Artocarpus tonkinensis. AB - Activity-guided fractionation of the n-butanol extract from the leaves of Artocarpus tonkinensis led to the isolation of the auronol glycosides maesopsin 4 O-glucoside (1), as well as the new alphitonin-4-O-glucoside (2). These structures were identified on the basis of MS and NMR spectroscopic data. The lymphocyte stimulation test showed both compounds having immunosuppressive activity. PMID- 15125578 TI - Diterpenes and norditerpenes from the roots of Dorystoechas hastata. AB - Five compounds, two diterpenoids ferruginol (1), and 6,7-didehydrosempervirol (2) and two norditerpenoids 17-hydroxycryptotanshinone (3), przewaquinone A (4) along with a new norditerpenoid cryptotanshinone 17beta-oic acid (5) have been isolated from the roots of Dorystoechas hastata Boiss. et Heldr. ex Bentham (Lamiaceae = Labiatae) growing wild in Southwestern Turkey (Antalya). PMID- 15125579 TI - Phenolic sodium sulphates of Frankenia laevis L. AB - Four new phenolic anionic conjugates have been isolated from the whole plant aqueous alcohol extract of Frankenia laevis L. Their structures were established, mainly on the basis of ESI-MS, 1D and 2D NMR spectroscopic evidence, as gallic acid-3-methyl ether-5-sodium sulphate, acetophenone-4-methyl ether-2-sodium sulphate, ellagic acid-3,3'-dimethyl ether-4,4'-di-sodium sulphate and ellagic acid-3-methyl ether-4-sodium sulphate. PMID- 15125580 TI - Triterpenoids from Salvia wagneriana. AB - From the exudate of Salvia wagneriana Polak, beside the known ursolic acid, two pentaoxygenated triterpenoids were isolated. Their structures were determined as 3-oxo-11alpha,19beta,20,22beta-tetrahydroxy-lupane (1) and 3beta,11alpha,19beta,20,22beta-pentahydroxy-lupane (2) using a combination of one and two-dimensional NMR techniques. PMID- 15125581 TI - Portable medicine chests in Serbia during the 19th and the first half of the 20th century. AB - Portable medicine chests could be considered as the oldest type of pharmacy in the 19th century Serbia. Historically their emergence could be associated with the appearance of the first physicians who kept post in Serbia. They were foreigners engaged on a contract to work during the reign of Prince Milos Obrenovic. According to the data preserved at the State Archive of Serbia (Chancellery of the principality, State Council, Home Office - Medical Department), medicine chests existed as late as the middle of the last century. They were an important source for health protection of the Serbian people as well as for distribution of drugs during the course of the 19th century. This article aims to analyse the content of these chests, based on the archive data records referring to the regular supervision of the medicine chests conducted every fourth year by the district physician. The legal aspect of keeping the portable medicine chest will be discussed as well. PMID- 15125582 TI - 3-Alkyl- and 3-aryl-7H-furo[3,2-g]chromen-7-ones as blockers of the voltage-gated potassium channel Kv1.3. AB - 3-Alkyl- and 3-aryl-7H-furo[3,2-g]chromen-7-ones were synthesised and studied for their effects on voltage-gated K+ and Na+ channels of the neuroblastoma cell-line N1E-115 and K+ channels of L-929 mouse-fibroblasts, stably transfected with mKv1.3. All furocoumarins tested showed Kv channel blocking activities, the most potent one in a half-blocking concentration of 0.7 microM. PMID- 15125583 TI - Synthesis and anti-inflammatory effect of lipophilic derivatives of threo-DL phenylserine in rat experimental edema. AB - Novel N- and O-acyl derivatives of threo-DL-phenylserine containing lipophilic long chain carboxylic or aryl(sulfon)amide groups were synthesized. Anti inflammatory activity in carrageenin-induced paw edema model in rats was studied. Compound 10 had the most expressed anti-inflammatory effect over 24 h. PMID- 15125584 TI - Identification and determination of invertase secreted by tomato cells. AB - A simple and rapid procedure for the identification and determination of extracellular invertase from a culture medium of tomato cell suspension cultures is described. Sucrose was used as substrate for the determination of the extracellular and intracellular activities of the enzyme. The culture medium (without cells) was used for identification and determination of extracellular enzyme activity. Intracellular activity was estimated from the cell suspension. PMID- 15125585 TI - The choice of lipids and surfactants for injectable extravenous microspheres. AB - Suspensions of lipid microspheres sizing from 1 to 30 microm, whose fluidity and lipid/surfactant composition is suitable for parenteral administration were developed. None of the formulations prepared with Precirol (palmitostearate), as the only lipid, was physically stable during storage, because liquid suspensions formed semisolid gels within one week. Stable 10% (w/w) suspensions of lipid microspheres were produced using saturated triglycerides in combination with medium chain unsaturated triglycerides (Miglyol) as lipids and polysorbate 80 (2% w/w) as a surfactant. PMID- 15125586 TI - Zinc sulphate release and morphology of matrices prepared for the individual therapy of Wilson's disease. AB - Hydrophobic zinc sulphate wax matrices with different drug loadings were prepared for the individual hospital therapy of Wilson's disease. The drug release parameters, scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS) of the samples were analysed. The release mechanisms from matrices of 75% and 80% w/w zinc sulphate loadings were described with good correlation by the semi-empirical Fikkian diffusion based release model. Besides the zinc sulphate diffusion through the pores of the wax matrices, the parallel diffusion of zinc sulphate from the matrix surface is dominant in the case of samples of 83% and 90%w/w drug loadings. The combination of SEM and EDS analysis visualizes the morphology of the matrices and the related composition thus explaining the differences in the release characteristics. PMID- 15125587 TI - Anti-inflammatory and antinociceptive activities of Lippia nodiflora Linn. AB - The methanolic extract of leaves of Lippia nodiflora Linn. was tested for its anti-inflammatory and antinociceptive activities. The extract showed a significant (P < 0.001) anti-inflammatory activity comparable to phenylbutazone against carrageenin-induced paw edema in rats and a significant (P < 0.001) antinociceptive activity comparable to diclofenac sodium in acetic acid induced writhing in white albino mice. PMID- 15125588 TI - Influence of surfactants on the physical stability of solid lipid nanoparticle (SLN) formulations. AB - The choice of surfactant or surfactant mixtures at suitable concentrations contributes to the stability of solid lipid nanoparticles (SLN). In this study, it was found that 1.5% TegoCare 450 was the most effective stabilizer for the Witepsol E85 SLN dispersion compared to Tween 80, Tyloxapol and Pluronic F68 according to the data obtained from differential scanning calorimetry (DSC), zeta potential (ZP) measurements and particle size analysis. PMID- 15125589 TI - The relationship between salivary secretion rate and masticatory efficiency. AB - In order to elucidate the relationship between the salivary secretion rate and masticatory efficiency, experimental hypo- and hyper-salivation were produced by the administration of atropine sulphate presenting an anticholinergic effect and pilocarpine hydrochloride having a muscarine effect orally in 10 healthy fully dentates. To confirm the pharmaceutical effect of these drugs, the unstimulated whole salivary secretion rate during 10 min, and masticatory efficiency using the sieve method were measured before and after medication. The unstimulated whole salivary secretion rate during 10 min decreased significantly by the administration of atropine sulphate (P < 0.05), and increased significantly by pilocarpine hydrochloride (P < 0.01). The masticatory efficiency after atropine sulphate medication was significantly lower than that before (P < 0.01). The increase in salivary secretion by pilocarpine hydrochloride did not lead to a higher masticatory efficiency. The evidence supports the understanding that saliva plays an important role in masticatory function. PMID- 15125590 TI - Time series analysis of jaw muscle contraction and tissue deformation during mastication in miniature pigs. AB - Masticatory muscle contraction causes both jaw movement and tissue deformation during function. Natural chewing data from 25 adult miniature pigs were studied by means of time series analysis. The data set included simultaneous recordings of electromyography (EMG) from bilateral masseter (MA), zygomaticomandibularis (ZM) and lateral pterygoid muscles, bone surface strains from the left squamosal bone (SQ), condylar neck (CD) and mandibular corpus (MD), and linear deformation of the capsule of the jaw joint measured bilaterally using differential variable reluctance transducers. Pairwise comparisons were examined by calculating the cross-correlation functions. Jaw-adductor muscle activity of MA and ZM was found to be highly cross-correlated with CD and SQ strains and weakly with MD strain. No muscle's activity was strongly linked to capsular deformation of the jaw joint, nor were bone strains and capsular deformation tightly linked. Homologous muscle pairs showed the greatest synchronization of signals, but the signals themselves were not significantly more correlated than those of non-homologous muscle pairs. These results suggested that bone strains and capsular deformation are driven by different mechanical regimes. Muscle contraction and ensuing reaction forces are probably responsible for bone strains, whereas capsular deformation is more likely a product of movement. PMID- 15125591 TI - Single tooth bite forces in healthy young adults. AB - The assessment of bite forces on healthy single tooth appears essential for a correct quantification of the actual impact of single implant oral rehabilitations. In the present study, a new single tooth strain-gauge bite transducer was used in 52 healthy young adults (36 men, 16 women) with a complete permanent dentition. The influences of tooth position along the dental arch, of side, and of sex, on maximum bite force were assessed by an ANOVA. No significant left-right differences were found. On average, in both sexes the lowest bite force was recorded on the incisors (40-48% of maximum single tooth bite force), the largest force was recorded on the first molar. Bite forces were larger in men than in women (P < 0.002), and increased monotonically along the arch until the first or second permanent molar (P < 0.0001). The present data can be used as reference values for the comparison of dental forces in patients. PMID- 15125592 TI - Laser Doppler flow investigation of fractured permanent maxillary incisors. AB - Laser Doppler flowmetry (LDF) is a non-invasive method to assess pulpal blood flow (PBF). Dental injury has been associated with losses of pulpal sensibility. The purpose of this study was to assess the outcome of splint therapy on tooth fracture type-related PBF values. In 15 trauma patients, a single maxillary incisor treated by repositioning and splinting was investigated by LDF to assess local PBF values. Perfusion units were taken in four sessions, on the day of splint removel, and 12, 24 and 36 weeks after splint removal. Analysis of the tooth fracture type-related PBF measurements revealed root fractures to be associated with a significant decrease (P < 0.05) and uncomplicated crown fractures to be associated with a significant increase in PBF values (P < 0.05), while complicated crown fractures showed no significant difference between the session-related values (P > 0.05). The main findings of this study suggest splint therapy of root fractures of the central maxillary incisor to be associated with a short- and long-term decrease in PBF values. The LDF may become useful in the detection of transient ischaemic episodes and the identification of teeth at risk for adverse sequelae such as avascular necrosis and tissue loss. PMID- 15125593 TI - Computer-assisted morphological analysis of dental casts with maxillary prognathism and reversed occlusion. AB - The morphologic characteristics of the dentition with maxillary prognathism and reversed occlusion were examined by a computer-assisted dental cast analysing system. Dental casts with normal occlusion, maxillary prognathism and reversed occlusion were selected and measured by a 3D shape measuring system. The dental arches and anteroposterior occlusal curves were approximated numerically by the polynomial expression with a fourth order and second order coefficients, respectively. The coefficients were analysed statistically. Maxillary dental arches with mandibular prognathism showed a more acuminate shape (V-shape) with a significant larger second order coefficient, while arches with reversed occlusion showed more angulate shapes (U-shape) with significantly smaller second order coefficients compared with arches with normal occlusions. However, a notable difference in the shape of the mandibular dental arches was not observed. The mandibular dental arches with maxillary prognathism were positioned backward relative to maxillary dental arches, while the arches with reversed occlusion were positioned forward. The anteroposterior occlusal curves with maxillary prognathism showed larger curvatures in maxilla and mandible, while the curves with reversed occlusion showed smaller curvatures in mandible comparing with the curves with normal occlusion. These results suggest that the present method could offer a useful and objective examination technique for the diagnosis of malocclusion. PMID- 15125594 TI - The influence of the accuracy of the intermaxillary relations on the use of complete dentures: a clinical evaluation. AB - The aim of this study was to investigate possible associations between the accuracy of intermaxillary relations and complete denture usage 3 months and 3 years after their insertion. A total of 600 patients were provided with new complete dentures. Willis gauge and Woelfel's method were used to assess the quality of the existing complete dentures and the use of these dentures. Three years after insertion the remaining 250 patients took part in a follow-up examination. Three months post-insertion, significant relationships were found between adequate interocclusal rest space or quality of registration of centric relation and daily wearing of complete dentures and between quality of registration of centric relation and use of dentures for eating. Three years after insertion the adaptation to denture wearing was generally satisfactory. However, no significant relationship was observed between the accuracy of intermaxillary relations and complete denture usage. PMID- 15125595 TI - Rationalization of quantitative tooth surface loss data for epidemiological research. AB - Recent UK National Surveys have revealed a high prevalence of tooth surface loss (TSL) because of erosion in Children and Adolescents. Although digital surface mapping and surface matching techniques may be used to quantify its progression with time, reporting TSL of individuals as a function of either mean depth loss or volume loss, this can be inadequate when transferred for epidemiological analysis. For example, a tooth displaying multiple regions of depth loss may deserve to be distinguished from a tooth with a single localised area of erosion, although the total volume change may be the same. A potential solution, explored here, is to use both the quantitative wear data and colour coded surface representation plots generated by such methods to arrive at a single categorical wear score [1 (< or = 5% of surface exhibits TSL) to 5 (> or = 51% of surface exhibits TSL)]. Two examiners independently categorized, on two separate occasions, the TSL of 53 maxillary incisors (26 subjects observed at baseline and 9 months). Their performance was assessed for intra- and inter-examiner agreement by; (i) calculating the percentage of agreement, (ii) a Wilcoxon matched-pairs signed rank test. Thereafter, in cases of disagreement a consensus score was allocated and a regression analysis of this versus the numerically derived percentage of the surface that had undergone change was carried out. Intra- and inter-examiner agreement was 100 and 88.7% respectively and for the inter examiner comparison P = 0.0456. The categorical scores and the quantitative wear data correlated linearly (R = 0.82). It is concluded that the method used to rationalize the quantitative TSL data is both reproducible and reflects the quantitative data. Its use should be helpful in epidemiological TSL studies but continued vigilance is required in cases of examiner disagreement. PMID- 15125596 TI - The immunohistochemical distribution of vimentin in human temporomandibular joint samples. AB - The aim of this investigation was to evaluate the immunohistochemical distribution of vimentin in the temporomandibular joint (TMJ) and to compare it with the control specimens. Immunohistochemical distribution in the disc and synovial membrane in 30 human TMJ (internal derangement of TMJ, n = 20; and control, n = 10) was studied immunohistologically using paraffin-embedded tissue and specific anti-human vimentin monoclonal antibody. Vimentin expression was distributed in chondrocyte-like cells, synovial cells and endothelial cells. There was an obvious distinction of vimentin immunoreactivity between the control specimens and internal derangement cases, in the posterior and/or anterior loose connective tissues. In particular, intensive vimentin expression was detected in the hypertrophic synovial membrane of internal derangement cases. The findings of the present study suggest that vimentin might be an important marker of pathological hypertrophy of the synovial membrane and/or connective tissue with internal derangement of TMJ. PMID- 15125597 TI - Comparison of clinical and magnetic resonance imaging diagnoses in patients with TMD history. AB - The aim of this study was to evaluate the diagnostic accuracy of a well-defined clinical examination for diagnosing anterior disc displacement with and without reduction. A series of 40 patients with temporomandibular disorder (TMD) history were examined according to well-defined routine criteria. This examination included detailed history-taking, standardized clinical head and neck examination that included measurements of the range of motion, palpation of the temporomandibular joints and muscles of mastication for pain and auscultation of joint sounds. Magnetic resonance images of the joints were used as 'gold standard'. Diagnostic accuracy of the clinical examination was 83% for determining normal disc-condyle relationship, 72% for diagnosing anterior disc displacement with reduction, and 81% for diagnosing anterior disc displacement without reduction. Our results suggest that anterior displacement of the disc can be diagnosed with considerable accuracy using a well-defined clinical examination only. Therefore, we conclude that not all patients with TMD symptoms require magnetic resonance imaging examination before treatment. PMID- 15125598 TI - Effect of home bleaching agents on the microhardness of human enamel and dentin. AB - In this study, the effects of home bleaching agents that contains carbamide peroxide (in different concentrations such as 10 and 15%) on the surface hardness of human enamel and dentine were evaluated. The buccal surfaces of 90 recently extracted human mandibular anterior teeth were used. The specimens in the dentine group were ground to expose of dentine surface. Then, samples in both enamel and dentine group were randomly divided into six main groups with 15 in each group. In the group 1 and 4, 15% carbamide peroxide was applied to the enamel and dentine surfaces and then the teeth were left in an incubator at 37 degrees C and 100% humidity for 4 h first, then left 28 h after repeating the same procedures seven times. The teeth were treated with 10% carbamide peroxide in groups 2 and 5 and distilled water in groups 3 and 6 (control groups), using the same procedures. Vicker's microhardness of enamel and dentine were assessed on each tooth before and after each treatment period (4 and 28 h). The hardness of control group specimens was also obtained after 4 h in distilled water and then after 28 h. For statistical analysis one-way ANOVA and post-hoc Tukey tests were used. For both 10 and 15%, there were no statistically significant differences between the untreated control specimens and the specimens treated with the bleaching materials for enamel and dentine at any given measurement time (P > 0.05). Nevertheless, in vivo use of bleaching agents on tooth hard tissues requires further analysis. PMID- 15125599 TI - Celluloid strip-finished versus polished composite surface: difference in surface discoloration in microhybrid composites. AB - The aim of this study was to compare the surface discoloration of polished and celluloid strip-finished composite surfaces in microhybrid type composites and to determine whether or not the discoloration of a celluloid strip-finished composite surface is reduced if oxygen inhibition on the surface can be prevented. The composite surfaces were celluloid-strip finished (group 1), polished (group 2), or celluloid-strip finished under nitrogen gas purging (group 3). Z100, Spectrum, and Aelitefil were used as the test materials. After each surface treatment, the samples were stored in 37 degrees C distilled water for 24 h and placed in a disclosing solution (0.2% Erythrosin, pH 7.0) for 7 days in the dark. A computer controlled spectrophotometer was used to determine the CIELAB co ordinates (L*, a*, b*). For each composite, the degree of discoloration among the groups was compared with a one-way ANOVA test. A Dunnett's t-test was then performed to compare the colour change in group 1 with that of group 2 or group 3. In all materials, there were no statistical differences in the surface discoloration between group 1 and group 2. In the Aelitefil and Spectrum test materials, the samples in group 3 showed a lower level of discoloration than the those in group 1. In Z100, there was no statistical difference. In microhybrid composites, there was no difference in surface discoloration between celluloid strip-finished and the polished surface. Nitrogen gas purging during the polymerization process on the celluloid strip-finished composite surface might be useful for reducing the discoloration celluloid strip-finished surface in microhybrid composites. PMID- 15125600 TI - Calcium phosphate formation on the phosphorylated dental bonding agent in electrolyte solution. AB - The aim of the present study was to study the mineral formation on a phosphorylated dental bonding agent using a mineralization inductive solution. Clearfil Photobond, which contained phosphate monomer, was cured by photo irradiation and heat treated, and was then immersed in Hanks' balanced salt solution (HBSS) with pH = 7.4 for 1, 3, 5, 7, 14, and 28 days at 37 degrees C. The white substances were deposited on the phosphorylated polymer, i.e. cured Photobond disk, after the immersion in HBSS. The white substances become visible after 3 days immersion. After 7 days immersion, surface of the phosphorylated polymer disk was almost covered with white substance layers. The measurement of white substances by means of X-ray diffraction, Fourier-transform infrared and electron probe microanalysis revealed that their main component was carbonate containing hydroxyapatite. Scanning electron microscopy pictures showed that a large number of globules of hydroxyapatite were fused together, and that each globule was composed of a group of numerous thin-film form flakes uniting and/or clustering together. The results obtained in this study concluded that the presence of phosphonic acid and phosphate group of phosphorylated dental bonding agent enhanced the nucleation and growth of hydroxyapatite crystals on its surface. PMID- 15125601 TI - Bending strength and depth of cure of light-cured composite resins irradiated using filters that simulate enamel. AB - This study evaluates the light-attenuating effects of enamel on the properties of light-cured restorative resins using simple experimental filters. Three filters were designed to replicate the light transmittance characteristics of 0.5, 1.0 and 1.5 mm thick human enamel. The bending strength, depth of cure, and levels of residual monomer for 12 shades of three commercial light-cured composite resins were examined. These resins were cured either using direct irradiation from a light source or irradiation through one of the filters. For all materials, the bending strength and depth of cure of specimens irradiated through a filter were lower and the levels of residual monomer were higher than those found in specimens irradiated directly. The results indicate that the light-attenuating effect of enamel reduces the polymerization efficiency, resulting in poorer mechanical properties of light-cured composite resins. PMID- 15125602 TI - A reappraisal of the incremental packing technique for light cured composite resins. AB - The cuspal flexure caused by a direct placement composite resin was measured using a technique that did not interfere with cuspal movement. Twenty upper premolar teeth with mesial-occlusal-distal (MOD) cavity preparations were restored using a posterior composite resin. Ten teeth were restored using a bulk packing technique and 10 were restored using three 'U' shaped bucco-lingual increments. Total cuspal movement was measured 1 h following the initiation of curing. Bulk placement of the composite resin produced slightly more cuspal movement (12.5 microm) compared with the incremental placement of composite (11.3 microm). This difference was not statistically significant. The technique of incremental packing to reduce cuspal flexure is therefore questioned. PMID- 15125603 TI - The effect of washing water on bonding to etched enamel. AB - There is current concern about bacterial contamination of dental unit waterlines. This research hypothesized that the presence of increasing concentrations of bacteria in water used to wash etched enamel would result in a corresponding decrease in both shear bond strength (SBS) and critical surface tension (gammaC) of enamel. A further hypothesis was made that there is a correlation between SBS and gammaC. The effect of 3.5 ppm iodine in the water as a bacteriostatic agent was also assessed. Five groups of 10 samples of bovine enamel were etched, washed, and a resin composite bonded to them. The control group was washed with distilled water. Another group was washed with the dilute iodine solution. The remaining three groups used a different concentration of Escherichia coli DH5alpha as follows (in cfu mL(-1)): group 1: 10(2); group 2: 10(4); group 3: 10(6). Shear bond strength data were measured on an Instron testing machine at a crosshead speed of 1 mm min(-1). Adhesion data were (MPa): control: 24.6 +/- 6.0; with iodine: 20.8 +/- 2.7; group 1: 19.8 +/- 2.7; group 2: 13.5 +/- 3.0; group 3: 13.9 +/- 3.6. The F-test yielded a highly significant difference between control group, iodine group and group 1, compared with groups 2 and 3 (P < 0.0001). Tukey's Studentized Range Test was used for pairwise comparison testing between groups. Using a Cahn dynamic contact angle analyzer and linear regression analysis, the plots of surface tension versus costheta were extrapolated to costheta = 1 to give gammaC data for the control group and groups 1-3. In all cases reasonable linearity was observed (r2 > or = 0.87). Data (mN m(-1)) were: control group: 50.8; group 1: 45.1; group 2: 43.2; group 3: 39.5. The SBS and gammaC were then plotted against each other and linear regression analysis performed. It was concluded that increasing concentrations of bacteria in wash water decreased both SBS and gammaC and that a linear correlation (R2 = 0.84) was found between the values of these two parameters. PMID- 15125604 TI - Immediate resin-bonded bridgework: results of a medium-term clinical follow-up study. AB - A total of 21 immediate resin-bonded bridges were inserted under controlled and standardized clinical conditions and evaluated over a period of 35 months. During the evaluation period, two bridges debonded (9.5%). The debonded bridges were re cemented successfully. This low incidence of debonding indicates that this simple alternative procedure for the immediate replacement of a single unsaveable tooth was satisfactory. The subjects included in this study expressed their satisfaction with this form of treatment. PMID- 15125605 TI - Subclinical celiac disease in children: refractory iron deficiency as the sole presentation. PMID- 15125606 TI - Propionic acidemia with myelodysplasia and neutropenia in a Turkish child. PMID- 15125607 TI - Defining the appropriate dosage of folinic acid after high-dose methotrexate for childhood acute lymphatic leukemia that will prevent neurotoxicity without rescuing malignant cells in the central nervous system. AB - Neurotoxicity after the administration of methotrexate continues to worry physicians. However, inadequate folinic acid rescue is often not considered as a cause of this complication. To clarify whether adequate folinic acid rescue prevents methotrexate-induced neurotoxicity without reducing the cure rate in childhood ALL, published evidence that supported or refuted this claim was investigated. A literature search was conducted and the authors of the relevant studies were contacted. The published data supported the contention that neurotoxicity can be prevented by adequate folinic acid rescue even after very high doses of methotrexate. The safe minimum dose of folinic acid can be defined in terms of the dose of methotrexate given; the time to start of rescue is probably less important. There was no evidence that higher doses of folinic acid, such as those used after methotrexate in the treatment of osteosarcoma, rescue leukemia cells. No change in cure rate was found in relation to changes in scheduling or clinically relevant doses of folinic acid rescue. The accumulation of folinic acid in the cerebrospinal fluid did not seem to be of clinical relevance. No studies indicate that doses of folinic acid after high-dose methotrexate administration interfere with the killing of leukemia cells, nor that delaying the start of rescue beyond a certain point increases the antileukemic effect; neurotoxicity will, however, be increased. Review of current protocols that use low-dose folinic acid rescue and are associated with neurotoxicity is highly recommended. PMID- 15125608 TI - Fatal familial infantile myelofibrosis. AB - Malignant megakaryopoiesis can cause chronic or acute myelofibrosis through production of fibrogenic cytokines. Chronic myelofibrosis is a clonal disorder with marrow fibrosis, myeloid metaplasia, gross splenomegaly, and teardrop cells. Acute myelofibrosis differs by its aggressiveness, by the fact that it is more common in children, and by lack of organomegaly or anisopoikilocytosis. Surprisingly, in early childhood and infancy, splenomegaly and teardrop red cells become an important feature. Infantile myelofibrosis is a rare disease, except in Down syndrome. Familial occurrence of infantile myelofibrosis is exceedingly rare. The author describes an unfortunate family whose four consecutive children died of a very fulminant form of acute myelofibrosis during their first year of life. The fulminant nature of the disease, the degree of splenomegaly, and the prominence of anisopoikilocytosis were even more marked than in currently reported cases of infantile myelofibrosis. The mode of inheritance remained illusive. With two female children, sex-linked inheritance was not possible. It could not have been inherited in an autosomal dominant fashion with normal parents and with two normal children from the father's second marriage. A new autosomal dominant mutation in the germ cell of either parent is another possibility. Autosomal recessive inheritance remained a logical explanation, although such a high degree of disease presentation in a non-consanguineous marriage seems to put that possibility in question. PMID- 15125609 TI - Chromosome abnormalities may correlate with prognosis in Burkitt/Burkitt-like lymphomas of children and adolescents: a report from Children's Cancer Group Study CCG-E08. AB - Among pediatric non-Hodgkin lymphomas, the most frequent type is small noncleaved cell lymphoma (including Burkitt and Burkitt-like). Specific chromosome abnormalities are associated with prognosis in childhood acute lymphoblastic leukemia (ALL); however, chromosome abnormalities have not been evaluated for prognostic value in pediatric Burkitt and Burkitt-like lymphomas. For Children's Cancer Group protocol CCG-E-08 Etiologic Study of Non-Hodgkin Lymphoma in Childhood, 19 patients were enrolled with cytogenetic analysis of Burkitt or Burkitt-like lymphoma and simultaneously enrolled on treatment protocols CCG-503 or CCG-552. Pathology material and karyotypes at initial diagnosis underwent central review. Demographics included an age range of 2 to 14 years (median 8 years) and a male:female ratio of 14:5. All patients had advanced disease (stages III and IV, or ALL). Disease relapsed in five patients (event-free survival 74%, median follow-up 10.4 years). Chromosome abnormalities were identified in 18 patients (95%) including t(8;14)(q24.1;q32) in 12 (63%); t(8;22)(q24.1;q11.2) in 1 (5%); partial duplication of 1q in 7 (37%); and 13q32 abnormalities in 2 (11%). In patients who had relapses, in addition to the t(8;14)(q24. ;q32), two had abnormalities of 13q32 and two had partial duplication of 1q. CMYC translocations were absent in Burkitt-like lymphomas from all three patients. Burkitt and Burkitt-like lymphomas in children have a high frequency of chromosome abnormalities. Burkitt lymphoma abnormalities often involve CMYC translocations, usually a t(8;14)(q24.1;q32). Additional chromosome abnormalities that involved 13q32 and partial duplication of 1q were associated with poor prognosis. Burkitt like lymphomas were not associated with CMYC translocations. Further studies are warranted in larger cohorts of children and adolescents with Burkitt and Burkitt like lymphomas. PMID- 15125610 TI - Chemical and functional analysis of hydroxyurea oral solutions. AB - The primary hypothesis of the upcoming NIH-sponsored phase III infant hydroxyurea (BABY HUG) trial is that hydroxyurea can prevent chronic organ damage in infants with sickle cell anemia. Since hydroxyurea is currently commercially available only in capsules, a liquid formulation of hydroxyurea is needed for young patients. Hydroxyurea oral solutions were prepared by dissolving the contents of the capsules in water (room temperature or mildly heated) with vigorous stirring, filtering excipients, and adding flavored syrup to a final concentration of 100 mg/mL. Chemical stability was determined by measuring the hydroxyurea concentration using a standardized analytical colorimetric analysis, while functional stability was determined by measuring the inhibition of phytohemagglutinin-induced T lymphocyte proliferation. Hydroxyurea oral solutions prepared using room-temperature water had statistically equivalent spectrophotometric concentration and inhibition of T-lymphocyte proliferation for 3 to 6 months. Mild heating of the water to facilitate dissolution of the hydroxyurea capsule contents resulted in a reduced concentration and inhibitory activity of the preparations. Hydroxyurea oral solutions (100 mg/mL) prepared and maintained at room temperature have chemical and functional stability for several months. Hydroxyurea oral solutions prepared and dispensed monthly are suitable for use in the upcoming infant BABY HUG trial. PMID- 15125611 TI - Umbilical cord blood transplantation in Chinese children with beta-thalassemia. AB - To evaluate factors affecting outcome of sibling umbilical cord blood transplantation in Chinese children with thalassemia. The authors conducted a retrospective review of all patients undergoing such transplants in a single institution. Nine children with thalassemia major were diagnosed at a median age of 12 months. They received irregular blood transfusions and suboptimal iron chelation therapy before transplant. Sibling cord blood transplant was performed at a median of 5.5 years (range 3.5-10 years). Six donors were HLA-identical; three were one- to three-antigen mismatched. The mean number of nucleated cells infused was 6.6 x 10(7)/kg (range 3.4-12.7); the mean number of CD34+ cells infused was 3.8 x 10(5)kg (range 0.6-11.7). Seven patients had engraftment of donor cells. The median number of days to achieve a neutrophil count of > 0.5 x 10(9)/L was 19 days (range 10-25); the median number of days to achieve a platelet count of > 20 x 10(9)/L was 33 days (range 19-63). Of the six patients who received HLA-identical transplants, one developed grade 2 and two developed grade 1 acute graft-versus-host disease. Two of the three patients receiving mismatched cord blood did not achieve engraftment, and the other one engrafted but developed grade 4 acute graft-versus-host disease. Two patients subsequently developed secondary graft rejection and had autologous marrow regeneration before day 60 posttransplantation. With a median follow-up of 49 months (range 38-64), eight patients survived but only four were transfusion-independent. Umbilical cord blood transplant appears to have a higher chance of nonengraftment and secondary rejection. A more intensive immunosuppressive conditioning regimen may be required. PMID- 15125612 TI - Second allogeneic hematopoietic stem cell transplantation for juvenile myelomonocytic leukemia: case report and literature review. AB - Juvenile myelomonocytic leukemia (JMML) is a rare clonal myeloproliferative disease in young childhood. Hematopoietic stem cell transplantation (HSCT) is the only way to cure the disease, but relapse after HSCT remains a major cause of treatment failure. A 5-year-old girl with JMML, who had experienced a relapse after the first transplant, did not respond to donor lymphocyte infusion and withdrawal of immune-suppressing agents. She was successfully treated using a second transplant. Detailed reports from the English literature since 1988 relating to a total of 13 JMML patients undergoing a second transplant were reviewed. Seven of the 13 JMML patients (54%) were alive and disease-free, with a median follow-up of 53 months after the second transplant. Within the first 6 months following the initial transplant, 10 JMML patients suffered either autologous recovery (n = 6) or early relapse (n = 4). Seven of the 10 (70%) were alive, with a median survival period of 53 months after the second transplant. Six JMML patients underwent retransplantation within 6 months of the first transplant, with three of these (50%) alive at follow-ups of 24, 57, and 90 months after the second procedure. The authors conclude that a second transplant within 6 months may be worth considering for JMML patients who experience autologous recovery or earlier relapse after the first transplant. PMID- 15125613 TI - Bacillus cereus central line infection in an immunocompetent child with hemophilia. AB - Bacillus species are increasingly recognized as pathogens in immunocompromised patients. The authors report a case of Bacillus cereus infection of a central line in an immunocompetent patient with hemophilia, which required line removal for complete cure. PMID- 15125614 TI - Sweet syndrome developing during treatment with all-trans retinoic acid in a child with acute myelogenous leukemia. AB - Acute febrile neutrophilic dermatosis (Sweet syndrome) has been reported in a few adults receiving all-trans retinoic acid for acute promyelocytic leukemia. The authors report a case of Sweet syndrome associated with the administration of all trans retinoic acid for acute promyelocytic leukemia in a pediatric patient. PMID- 15125615 TI - Allogeneic bone marrow transplantation in juvenile myelomonocytic leukemia without total body irradiation. AB - Allogeneic bone marrow transplantation (BMT) without a total body irradiation (TBI) conditioning regimen was investigated in children with juvenile myelomonocytic leukemia (JMML). Eight consecutive patients with JMML (n = 6) or monosomy 7 (n = 2) underwent BMT at a median age of 20 months. Donor source included fully matched related (n = 3), mismatched related (n = 2), or fully matched unrelated (n = 3). The conditioning regimen included busulfan, cyclophosphamide, and etoposide (VP16) (melphalan was substituted for VP16 in one patient). The first patient in the series underwent TBI. Graft-versus-host disease prophylaxis was with cyclosporin and methotrexate and in vivo T-cell depletion (Campath 1 g) for mismatched and unrelated transplants. Seven and two patients, respectively, received chemotherapy and splenectomy before BMT. At a median follow-up of 48 months after BMT, five patients remained in remission. The overall survival rate was 63% at 5 years. All deaths occurred in patients with refractory disease at the time of BMT. Allogeneic BMT without TBI appears to be effective therapy for JMML and avoids some of the potential late sequelae of TBI in preschool children. PMID- 15125616 TI - Cancer in Sotos syndrome: report of a patient with acute myelocytic leukemia and review of the literature. AB - Sotos syndrome is a rare congenital disorder that is associated with various malignancies, including acute lymphoblastic leukemia and lymphomas. The NSD1 gene haploinsufficiency is associated with this syndrome. The authors report a case of acute myeloid leukemia developing in a child with Sotos syndrome. He was treated with standard chemotherapy and achieved sustained remission. On review of the literature, it was found that most malignancies in Sotos syndrome occur in childhood. In conclusion, because of their increased risk of developing malignancy, patients with Sotos syndrome should be followed closely for signs and symptoms of both hematologic and nonhematologic malignancies, at least during childhood. PMID- 15125617 TI - Recurrent eosinophilic cystitis in a child with chronic granulomatous disease. AB - Eosinophilic cystitis is an uncommon disease in children, and its association with chronic granulomatous disease (CGD) has been previously reported in only five patients. In all those patients the disease showed either a self-limited benign course or a rapid response to corticosteroid treatment. The authors describe a child with X-linked CGD who developed eosinophilic cystitis with a recurrent course and difficult therapeutic management. The authors also discuss the pathogenesis of granuloma formation in CGD and review the literature for current therapies for these complications. PMID- 15125618 TI - Asymptomatic babesiosis in a child with hepatoblastoma. PMID- 15125619 TI - Hypopigmentation from imatinib mesylate (Gleevec). PMID- 15125620 TI - Methods to evaluate risks for composite end points and their individual components. AB - OBJECTIVE: Both randomized and observational studies commonly examine composite end points, but the literature on model development and criticism in this setting is limited. STUDY DESIGN AND SETTING: We examined approaches for evaluating heterogeneity in the effects of risk factors for different components of the end point, and determining the impact of heterogeneity on the ability to predict the composite end point. A specific example considered the composite cardiovascular disease end point in the Physicians' Health Study that occurred in 1,542 (myocardial infarction, n = 716; stroke, n = 557; cardiovascular death, n = 269) of 16,688 participants with complete information on baseline covariates. The strategy compared alternative polytomous logistic regression models assuming different effects of risk factors on components of the end point and a comparable logistic model assuming common effects. RESULTS: Likelihood ratio tests identified heterogeneity in the effects of age, alcohol consumption, and diabetes across components of the outcome, but comparability in the effects of other risk factors. However, a model assuming uniform effects explained over 90% of the log likelihood change in the best polytomous model, and the two models also performed similarly based on a comparison of ROC curves. CONCLUSION: The overall strategy may be helpful for evaluating the validity of a composite end point analysis and identifying heterogeneity in risk factors. PMID- 15125621 TI - Assessment of coverage rates and bias using double sampling methodology. AB - OBJECTIVE: In 1999, a survey of the health status of six rural central New York counties was performed using double sampling. The resulting impact of the methodology on health outcome prevalence estimation was assessed. STUDY DESIGN AND SETTING: Six counties were double sampled using methodology described by Neyman. Prevalence of health outcomes was estimated for the initial survey responders, nonresponders who subsequently responded after being offered a financial incentive, and both groups combined. RESULTS: Although both groups were similar on demographic variables, they differed significantly on several disease and health behavior prevalences. The failure to use double sampling would have resulted in overestimation of the prevalence of chronic disease by as much as 6.2% for females and 2.0% for males. CONCLUSION: Double sampling increased the generalizability of the results from approximately 35% of the population to over 70%, and decreased the bias in the estimation of many of the health end points. PMID- 15125622 TI - Validation of diagnostic codes within medical services claims. AB - OBJECTIVES: Few studies have attempted to validate the diagnostic information contained within medical service claims data, and only a small proportion of these have attempted to do so using the medical chart as a gold standard. The goal of this study is to determine the sensitivity and specificity of medical services claims diagnoses for surveillance of 14 drug disease contraindications used in drug utilization review, the Charlson comorbidity index and the Johns Hopkins Adjusted Care Group Case-Mix profile (ADGs). STUDY DESIGN AND SETTING: Diagnoses were abstracted from the medical charts of 14,980 patients, and were used as the "gold standard," against which diagnoses obtained from the administrative database for the same patients were compared. RESULTS: Conditions associated with drug disease contraindications with the exception of hypertension and chronic obstructive pulmonary disease (COPD) showed a specificity of 90% or higher. Sensitivity of claims data was substantially lower, with glaucoma, hypertension, and diabetes being the most sensitive conditions at 76, 69, and 64%, respectively. Each of the 18 disease conditions contained in the Charlson comorbidity index showed high specificity, but sensitivity was more variable among conditions as well as by coding definitions. Although ADG specificity was also high, the vast majority of ADGs had sensitivities of less than 60%. CONCLUSION: The administrative data was found to have diagnoses and conditions that were highly specific but that vary greatly by condition in terms of sensitivity. To appropriately obtain diagnostic profiles, it is recommended that data pertaining to all physician billings be used. PMID- 15125623 TI - Pharyngitis clinical prediction rules: effect of interobserver agreement: a MetroNet study. AB - OBJECTIVE: Pharyngitis clinical prediction rules improve Group A beta-hemolytic streptococcus (GABHS) diagnosis and decrease unnecessary antibiotic use, yet few studies have addressed clinician variability in assessment of sore throat signs and symptoms. STUDY DESIGN AND SETTING: We conducted a cross-sectional study in which two clinicians examined each of 200 adult sore throat patients. Each patient had a rapid GABHS antigen test. Clinicians were blinded to each other's assessment and to the rapid antigen result. Interobserver agreement was estimated using a kappa coefficient. Effect of agreement on sensitivity, specificity, and hypothetic rapid antigen testing and antibiotic prescribing was determined for two clinical prediction rules. RESULTS: We found moderate inter-rater reliability on sore throat history and physical assessments. Clinician agreement was associated with significantly fewer hypothetic rapid antigen tests performed. CONCLUSION: Interobserver agreement enhances the utility of pharyngitis clinical prediction rules. Medical school and residency training should focus on correct assessment of history and physical examination components used in GABHS clinical prediction rules. Correct assessment will result in less GABHS testing and antibiotic prescriptions for sore throat patients. PMID- 15125624 TI - Selection bias found in interpreting analyses with missing data for the prehospital index for trauma. AB - OBJECTIVE: To evaluate the effects of missing data on analyses of data from trauma databases, and to verify whether commonly used techniques for handling missing data work well in theses settings. STUDY DESIGN AND SETTING: Measures of trauma severity such as the Pre-Hospital Index (PHI) are used for triage and the evaluation of trauma care. As conditions of trauma patients can rapidly change over time, estimating the change in PHI from the arrival at the emergency room to hospital admission is important. We used both simulated and real data to investigate the estimation of PHI data when some data are missing. Techniques compared include complete case analysis, single imputation, and multiple imputation. RESULTS: It is well known that complete case analyses and single imputation methods often lead to highly misleading results that can be corrected by multiple imputation, an increasingly popular method for missing data situations. In practice, unverifiable assumptions may not hold, meaning that it may not be possible to draw definitive conclusions from any of the methods. CONCLUSION: Great care is required whenever missing data arises. This is especially true in trauma databases, which often have much missing data and where the data may not missing at random. PMID- 15125625 TI - Osteoporosis health belief scale: minor changes were required after telephone administration among women. AB - OBJECTIVE: The Osteoporosis Health Belief Scale (OHBS) is a 42-item scale designed to measure general health motivation, perceived susceptibility to and seriousness of osteoporosis, and beliefs about calcium intake and exercise in preventing and treating osteoporosis. The purpose of this study was to examine the psychometric properties of the OHBS by telephone administration among older women. STUDY DESIGN AND SETTING: A convenience sample of 425 women aged 61-93 years participating in a longitudinal arthritis study was recruited by telephone. Item clarity was evaluated and 22 additional items (6 reworded, 16 from other questionnaires) were considered to supplement or replace existing scale items. Multitrait scaling techniques and exploratory factor analysis were used to test scale structure. Construct validity was tested based on theoretical hypothesis between OHBS subscale scores and participant characteristics. RESULTS: A few modifications to the OHBS scale were suggested, reducing the scale by five items (two redundant, three did not load), rewording one item and moving on item to a different subscale. The modified 37-item OHBS had a seven-factor uncorrelated solution explaining 48% of the model variance with internal consistency ranging from 0.73 to 0.88. CONCLUSION: Relatively minor changes to the OHBS results in reduced redundancy and improved internal structure of the scale for telephone administration among women over 60 years of age. Further examination is recommended to confirm these findings. PMID- 15125626 TI - Patient preferences for expectant management vs. surgical evacuation in first trimester uncomplicated miscarriage. AB - OBJECTIVE: Expectant and surgical management are widely accepted treatment options in case of a miscarriage. In the absence of differences in complications, the choice between both options can be based on patient preferences. STUDY DESIGN AND SETTING: We studied treatment preferences for future miscarriages in women with a miscarriage randomized to expectant or surgical treatment or managed according to their own choice. RESULTS: Data from 136 patients were analyzed. Women randomized either to expectant or surgical management opted for the allocated treatment in future in 55 and 74%, respectively. Of the women randomized to expectant management and with a successful spontaneous loss, 71% opted again for this treatment. Women who were managed according to their own treatment choice, held on to their initial treatment preference (expectant vs. surgical management; 84 and 88%, respectively). Preferences after treatment were strong. CONCLUSION: A strong treatment preference should be taken into account in the counseling process. Women without a preference should be well informed on all medical aspects of the two options in order to facilitate informed-shared decision-making. PMID- 15125627 TI - Natural history and prognostic indicators of sciatica. AB - OBJECTIVE: Sciatica is thought to have a good clinical outcome, but in fact, its natural history is not well known. STUDY DESIGN AND SETTING: We studied a prospective cohort of 3,164 workers from the French national electricity and gas company. In this cohort, 622 subjects suffered from sciatica in 1991. The predictive factors of the persistence or recurrence of sciatica after 2 years were identified by multivariate analysis (logistic regression). RESULTS: Of the 622 subjects with sciatica in 1991, 55% still reported its symptoms in 1993 and 53% in 1995. Of those who had recovered from sciatica in 1993, 61% still had low back pain and 27% of them long-lasting low back pain in 1993. The factors predictive of the persistence or recurrence of sciatica in 1993, identified by multivariate analysis, were: driving at least 2 hr/day, carrying heavy loads at work, a high level of psychosomatic problems, and sciatica symptoms the year before study inclusion. CONCLUSION: Recovery from sciatica is less frequent than expected. Attention should be given to occupational and personal factors associated with persistence or recurrence of sciatica. PMID- 15125628 TI - Perception of prognostic risk in patients with multiple sclerosis: the relationship with anxiety, depression, and disease-related distress. AB - OBJECTIVE: The aim of the study was to investigate the impact of perception of prognostic risk on anxiety, depression, and disease-related distress in patients with multiple sclerosis (MS). STUDY DESIGN AND SETTING: Perceived risk and perceived seriousness of the 2-year, 10-year, and lifetime prognosis of wheelchair dependence, disability status, anxiety, depression, and disease related distress were assessed in 101 patients. Distress was measured as the intrusion and avoidance of MS-related thoughts and feelings. RESULTS: Patients with higher perceptions of 2-year, 10-year, or lifetime risk were bothered by more intrusion of MS-related thoughts and feelings. Only higher perception of the 2-year risk of wheelchair dependence was significantly related with higher levels of anxiety, depression, and avoidance. Similarly, higher perception of the seriousness of wheelchair dependence was consistently associated with more intrusion and avoidance, but only perceived seriousness of the 2-year prospect of wheelchair dependence was significantly correlated with anxiety and depression. All relations were independent of clinically assessed disability status. CONCLUSION: Perceptions of the short-term risk and seriousness of wheelchair dependence were significantly related to anxiety, depression, and disease-related distress in patients with MS. These findings underscore the importance of informing patients with chronic disorders about the short-term prognosis of important long-term consequences of disease. PMID- 15125629 TI - The longitudinal relation between chronic diseases and depression in older persons in the community: the Longitudinal Aging Study Amsterdam. AB - OBJECTIVE: The purposes of this study were to examine the differences in influence of various chronic diseases on depressive symptomatology over time and to determine whether there were differences in such influence depending on physical limitations and time of onset of disease. STUDY DESIGN AND SETTING: Data for this study were obtained from the Longitudinal Aging Study Amsterdam. Two thousand two hundred eighty-eight respondents (age 55-85) were included and followed for a maximum of 6 years. Depressive symptoms (using the CES-D scale), the presence of seven frequently occurring chronic diseases, physical limitations, and sociodemographic variables were assessed by structured interviews. Generalized estimating equation models were estimated for each disease and compared with each other. RESULTS: Lung disease, arthritis, cardiac disease, and cancer were all positively associated with increased depressive symptoms over time. Stroke was associated with depressive symptoms, but these associations were not found when adjusted for physical limitations. For atherosclerosis and diabetes mellitus only weak or no associations with depressive symptoms were found. Recent onset of disease resulted in less strong associations for in cancer, lung disease, and arthritis. CONCLUSION: The results demonstrate that the level of depressive symptoms varies across type of chronic disease. In cardiac disease, arthritis, cancer, and lung disease increased depressive symptoms could not be attributed to physical limitations, but in stroke the association found with depressive symptoms was to a large extent attributable to physical limitations. PMID- 15125630 TI - Serum albumin and risk of stroke, coronary heart disease, and mortality: the role of cigarette smoking. AB - OBJECTIVE: Lower levels of serum albumin are associated with increased risk of all-cause and cardiovascular mortality as well as with coronary heart disease and stroke incidence. These relationships have been examined with specific focus on the role of cigarette smoking. STUDY DESIGN AND SETTING: A prospective study of 7,690 British men aged 40-59 years, with 16.8 years mean follow-up. RESULTS: Cigarette smoking was strongly and inversely associated with serum albumin concentrations that reverted to levels seen in never smokers after 5 years' cessation. Only in current and former smokers were there significant inverse relationships between serum albumin and risk of major CHD and stroke events even after adjustment for potential confounders. Only in current smokers was a significant inverse relationship seen between serum albumin and mortality from cardiovascular disease, cancer, and all causes. CONCLUSION: The inverse association between serum albumin concentration and disease outcome appears to be related to the effects of cigarette smoking on serum albumin concentration. PMID- 15125631 TI - Trends in old-age mortality in seven European countries, 1950-1999. AB - OBJECTIVE: Different from the general observed decline in old-age mortality, for The Netherlands and Norway there have been reports of stagnation in the decline since the 1980s. We detect periods of stagnation in recent old-age mortality trends, and explore for which causes of death the recent stagnation is most apparent. STUDY DESIGN AND SETTING: We applied Poisson regression analysis to total and cause-specific mortality data by age (80+), period (1950-1999), and sex for seven European low-mortality countries. RESULTS: We found large heterogeneity in the pace of decline in the countries under investigation, with periods of stagnation being widespread. In the 1980s and 1990s, stagnation was observed in Denmark, The Netherlands, and Norway (males). Continued mortality decline was observed especially in France. Although smoking has had a marked influence on the trends in old-age mortality, the role of smoking in the recent stagnation seems only modest and restricted to Norway. Mortality from cardiovascular diseases showed important crossnational variations in the pace of decline. Mortality from diseases specifically related to old age increased recently in all countries, except France. CONCLUSION: Old-age mortality seems highly plastic and susceptible to many factors, with both favorable and unfavorable effects on trends over time. PMID- 15125632 TI - Patient satisfaction: is it a measure for the outcome of care or the process of care? PMID- 15125633 TI - Goodbye, number needed to treat? PMID- 15125634 TI - Cohesin component dynamics during meiotic prophase I in mammalian oocytes. AB - Cohesins are chromosomal proteins that form complexes involved in the maintenance of sister chromatid cohesion during division of somatic and germ cells. Three meiosis-specific cohesin subunits have been reported in mammals, REC8, STAG3 and SMC1 beta; their expression in mouse spermatocytes has also been described. Here we studied the localization of different meiotic and mitotic cohesin components during prophase I in human and murine female germ cells. In normal and atretic human fetal oocytes, from leptotene to diplotene stages, REC8 and STAG3 colocalize in fibers. In murine oocytes, SMC1beta, SMC3 and STAG3 are localized along fibers that correspond first to the chromosome axis and then to the synaptonemal complex in pachytene. Mitotic cohesin subunit RAD21 is also found in fibers that decorate the SC during prophase I in mouse oocytes, suggesting a role for this cohesin in mammalian sister chromatid cohesion in female meiosis. We observed that, unlike human oocytes, murine synaptonemal complex protein SYCP3 localizes to nucleoli throughout prophase I stages, and centromeres cluster in discrete locations from leptotene to dictyate. At difference from meiosis in male mice, the cohesin axis is progressively lost during the first week after birth in females with a parallel destruction of the axial elements at dictyate arrest, demonstrating sexual dimorphism in sister chromatid cohesion in meiosis. PMID- 15125635 TI - Cytological evidence for triploid males and females in the bumblebee, Bombus terrestris. AB - The presence of both triploid males and females in Bombus terrestris was detected by distinct chromosome observation. These cytological features are novel among the higher Hymenopteran insects. We thus strictly applied the complementary sex determination (CSD) model previously proposed for Hymenopteran insects. Three out of 60 sibling queens that were mated with diploid males produced both triploid males and females, and founded colonies. The male to female ratio of the bees which emerged from the fertilized eggs of the queens was approximately 1 to 1. Thus we reconfirm that the sex in B. terrestris is determined by a single multi allelic locus. The body size of the triploid males was smaller than that of the diploid and haploid males. We found hatched eggs laid by one triploid female (worker). One of these developed into a 3rd instar larva, however most of the triploid individuals were sterile. PMID- 15125636 TI - Transcriptional repression mechanisms of nucleolus organizer regions (NORs) in humans and chimpanzees. AB - Polymorphisms related to transcriptional inactivation of nucleolus organizer regions (NORs) have long been described in many animals, particularly humans. However, the precise aetiology of such variations is not always clear. We conducted analyses to investigate the repression mechanisms in humans and chimpanzees using FISH (fluorescence in situ hybridisation) with 18S rDNA, Ag-NOR (silver nitrate) staining, C-banding, and the in situ nick translation technique with the HpaII restriction enzyme. Examination of 48 humans and 46 chimpanzees suggested that there are at least three different mechanisms that produce inactivation of NORs. These include: (1) elimination of rDNA; (2) DNA methylation: (3) gene silencing due to position effects induced by heterochromatin (C-bands) and/or telomeres. PMID- 15125637 TI - Molecular-cytogenetic characterization of the origin and the presence of pericentromeric euchromatin on minute supernumerary marker chromosomes (SMCs). AB - Small supernumerary marker chromosomes (SMCs) in human can be defined as additional centric chromosome fragments smaller than chromosome 20. For most small or minute SMCs a correlation with clinical symptoms is lacking, mostly due to problems in visualizing their euchromatic content. Recently we described two new molecular cytogenetic approaches for the comprehensive characterization of small SMCs, excluding those few cases with neo-centromeres. Minute SMCs, consisting preferentially of alpha-satellite DNA, are characterizable in one step by the centromere-specific multicolor FISH (cenM-FISH) approach. For further characterization of minute SMCs and eventually present euchromatic content, the recently developed centromere-near-specific multicolor FISH (subcenM-FISH) technique can be applied. These two approaches are highly informative and easy to perform, as demonstrated in the present report on the example of a prenatal case with a minute SMC derived from chromosome 3 cytogenetically described as min(3)(:p12.1 --> q11.2:). PMID- 15125638 TI - FAST-FISH with laser beam microdissected DOP-PCR probe distinguishes the sex chromosomes of Silene latifolia. AB - We present an improved FISH strategy for differentiating the sex chromosomes of the dioecious model plant, Silene latifolia. Fixed mitotic protoplasts were dropped on a polyethylene naphthalate membrane, the X or Y chromosomes were isolated using nitrogen laser beam microdissection, catapulted by laser pressure, and amplified by DOP-PCR. A modified FAST-FISH protocol based on a short hybridization time combined with a low concentration of probe was used. The success of this approach is demonstrated by the differential labeling of the X and Y chromosomes and it could represent a quick method for comparing organization of plant genomes. PMID- 15125639 TI - Construction, characterization and chromosomal mapping of bacterial artificial chromosome (BAC) library of Yunnan snub-nosed monkey (Rhinopithecus bieti). AB - We constructed a high redundancy bacterial artificial chromosome library of a seriously endangered Old World Monkey, the Yunnan snub-nosed monkey (Rhinopithecus bieti) from China. This library contains a total of 136 320 BAC clones. The average insert size of BAC clones was estimated to be 148 kb. The percentage of small inserts (50-100 kb) is 2.74%, and only 2.67% non-recombinant clones were observed. Assuming a similar genome size with closely related primate species, the Yunnan snub-nosed monkey BAC library has at least six times the genome coverage. By end sequencing of randomly selected BAC clones, we generated 201 sequence tags for the library. A total of 139 end-sequenced BAC clones were mapped onto the chromosomes of Yunnan snub-nosed monkey by fluorescence in-situ hybridization, demonstrating a high degree of synteny conservation between humans and Yunnan snub-nosed monkeys. Blast search against human genome showed a good correlation between the number of hit clones and the size of the chromosomes, an indication of unbiased chromosomal distribution of the BAC library. This library and the mapped BAC clones will serve as a valuable resource in comparative genomics studies and large-scale genome sequencing of nonhuman primates. The DNA sequence data reported in this paper were deposited in GenBank and assigned the accession number CG891489-CG891703. PMID- 15125640 TI - Expression pattern of X-linked genes in sex chromosome aneuploid bovine cells. AB - Expression of the X-inactive specific transcript (XIST) gene is a prerequisite step for dosage compensation in mammals, accomplished by silencing one of the two X chromosomes in normal female diploid cells or all X chromosomes in excess of one in sex chromosome aneuploids. Our previous studies showing that XIST expression does not eventuate the inactivation of X-linked genes in fetal bovine testis had suggested that XIST expression may not be an indicator of X inactivation in this species. In this study, we used a semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) approach on cultures of bovine cells with varying sex chromosome constitution (XY, XX, XXY and XXX) to test whether the levels of XIST expressed conform to the number of late replicating (inactive) X chromosomes displayed by proliferating cells in these cultures. Expression patterns of four X-linked genes, including hypoxanthine phosphorybosyl transferase (HPRT), glucose-6-phosphate dehydrogenase (G6PD), zinc finger protein locus on the X (ZFX). and 'selected mouse cDNA on the X' (SMCX), in all these cells were also tested. Results showed that XIST expression was significantly higher (p < 0.05) in XXX cells compared to XX and XXY cells and that G6PD. HPRT, and SMCX loci are subject to X inactivation. The significantly higher levels of ZFX expressed in XXX cells compared to XX and XXY cells (p < 0.05) confirmed that this bovine locus, as human ZFX, escapes X inactivation. However, the levels of XIST and ZFX expressed were not proportional to the X chromosome load in these cells suggesting that X-linked loci escaping inactivation may be regulated at transcription (or post-transcription) level by mechanisms that prevent gene specific product accumulation beyond certain levels in sex chromosome aneuploids. PMID- 15125641 TI - Expression of XIST sense and antisense in bovine fetal organs and cell cultures. AB - Untranslated RNAs transcribed from sense and antisense strands of a gene referred to as X-inactive specific transcript (XIST) play crucial roles in the genetic inactivation and condensation of one of the two X chromosomes in the somatic cells of female mammals. X inactivation is also thought to occur in mammalian male germ cells mainly based on the formation of a condensed structure referred to as a sex body or XY-body, during spermatogenesis. Molecular identity of the sex body, the roles of sense and antisense XIST RNAs in its formation, and the relevance of the sex body to spermatogenesis are not known. Here we report the results of our strand-specific RT-PCR approach to identify the amplicon detected in fetal bovine testes previously referred to as XIST and to test for sense/antisense expression in male and female organs and cell cultures of different sex chromosome constitution. Our results showed that the transcript detected consistently in male gonads and variably in somatic organs represents XIST antisense RNA and that XIST sense and antisense RNAs are co-expressed in female somatic tissues and cultured cells including cells of sex chromosome aneuploids (XXY and XXX). Our results, which differ from those of other investigators in this area, are discussed in the light of the recently reported differences in the expression pattern of murine Xist/Tsix loci and their structural and functional differences in different mammalian species. PMID- 15125642 TI - Application of AFLP technology to radiation hybrid mapping. AB - We have investigated the use of AFLP technology as a tool for the high throughput enrichment of Radiation Hybrid (RH) maps. The 3000 rad TM112 bovine RH panel was assayed with 37 EcoRI/TaqI AFLP primer combinations. The number of selective nucleotides used during PCR was increased to seven, to reduce the complexity of the AFLP profile and minimise the overlap between hamster and bovine bands co amplified from hybrid cell clones. Seven-hundred-forty-seven bovine AFLP bands were amplified that could be distinguished following electrophoresis. Repeatability was tested within and between laboratories on independent template preparations and an error rate of 1.3% found. Two-point linkage analysis clustered 428 AFLP fragments in 39 linkage groups of at least 4 markers. Multi point maps were constructed for 5 sample linkage groups. The study demonstrated that the AFLP approach could be used to rapidly screen for the most informative clones during panel construction and to increase the number of markers on RH maps, which could be useful for joining linkage groups formed by other markers. The use of AFLP markers as anchor points between existing RH maps and other physical maps, such as BAC contigs, is also discussed. PMID- 15125643 TI - Karyotype stability of the DT40 chicken B cell line: macrochromosome variation and cytogenetic mosaicism. AB - The DT40 transformed chicken B-cell line is an important and widely used vertebrate cell line. Knowledge of the 'wild-type' DT40 karyotype is a significant consideration for most research applications and for extrapolation of results to normal cells. Here we present data indicating that (1) modal karyotype differences exist between DT40 cultures from different sources, (2) DT40 cultures are cytogenetically mosaic, and (3) the mosaic features can change over time with media conditions influencing the degree and type of non-modal karyotypes that emerge. In addition to the previously reported trisomy for chromosome 2, a monosomy for chromosome 4 and a variant 4p- are described. The macrochromosomal variation and mosaic nature of the DT40 system should alert researchers using this important cell line to consider whether such variation will impact experimental results and interpretation. The results are discussed in regard to current knowledge of cytogenetic anomalies associated with chicken cancers. PMID- 15125644 TI - Pattern-based detection of different proteins using an array of fluorescent protein surface receptors. AB - A small library of porphyrin derivatives whose fluorescence emission changes on binding to protein surfaces has been developed as a protein "fingerprinting" array. When incubated with different proteins, the array yields a characteristic response, specific for every protein analyte. At the same time, this design allows for the rapid screening of the porphyrin library for the identification of high affinity protein surface ligands. PMID- 15125645 TI - Mapping of the binding site on pseudoazurin in the transient 152 kDa complex with nitrite reductase. AB - Nitrite reductase (NiR) catalyzes the reduction of nitrite to nitrite oxide as a part of the denitrification process. In Alcaligenes faecalis S-6, the copper protein pseudoazurin acts as electron donor to NiR. The binding surface of pseudoazurin involved in the formation of the 152 kDa complex with NiR has been determined by NMR using cross saturation from NiR to perdeuterated pseudoazurin. Due to the transient nature of the complex, saturation effects can be observed on the resonances of the unbound protein. The binding site comprises the hydrophobic area surrounding the exposed copper ligand His81, suggesting that this residue is important for efficient electron transfer. PMID- 15125646 TI - Enhanced production of functional proteins from defective genes. AB - Nonsense mutations are associated with a host of genetic disorders. The protein products encoded by genes containing these mutations are either truncated or completely missing. Recently, members of the aminoglycoside family of antibiotics have been found to induce the ribosome to read past these inappropriately inserted stop codons, albeit at extremely modest efficiencies. We report herein a strategy that enhances the production of functional proteins from the corresponding mutated genes. PMID- 15125647 TI - A ruthenium-grafted hydrotalcite as a multifunctional catalyst for direct alpha alkylation of nitriles with primary alcohols. AB - Treatment of a hydrotacite, Mg6Al2(OH)16CO3, with an aqueous solution of RuCl3.nH2O afforded a monomeric Ru(IV) species on the surface of the hydrotalcite. This novel Ru-grafted hydrotalcite (Ru/HT) efficiently catalyzed alpha-alkylation of nitriles with primary alcohols through the cooperative catalysis between the Ru species and the surface base sites. The catalyst system could be further extended for the one-pot synthesis of alpha,alpha-dialkylated phenylacetonitriles via the base-catalyzed Michael reaction of alpha-alkylated phenylacetonitrile with activate olefins. PMID- 15125648 TI - Facile one-pot synthesis of bifunctional heterodimers of nanoparticles: a conjugate of quantum dot and magnetic nanoparticles. AB - Sequential addition of sulfur and Cd(acac)2 into the colloid solution of FePt nanoparticles ( approximately 2.5 nm) under a reductive environment generates heterodimers of CdS and FePt with sizes of approximately 7 nm. The heterodimers exhibit both superparamagnetism and fluorescence, indicating that the discrete properties of the individual parts of the dimers are preserved. This simple methodology may lead to the production of large quantities of various heterostructures with tailored properties on the nanoscale. PMID- 15125649 TI - Hydrogen sorption in functionalized metal-organic frameworks. AB - Five porous metal-organic frameworks based on linking zinc oxide clusters with benzene-1,4-dicarboxylate, naphthalene-2,6-dicarboxylate, 4,5,9,10 tetrahydropyrene-2,7-dicarboxylate, 2,3,5,6-tetramethylbenzene-1,4-dicarboxylate, or benzene-1,3,5-tris(4-benzoate) were synthesized in gram-scale quantities to measure their hydrogen uptake properties. Hydrogen adsorption isotherms measured at 77 K show a distinct dependence of uptake on the nature of the link. At 1 atm, the materials sorb between 4.2 and 9.3 molecules of H2 per formula unit. The results imply a trend in hydrogen uptake with the number of rings in the organic moiety. PMID- 15125650 TI - Isolation and characterization of a monomeric cationic titanium hydride. AB - Methyl cations 1-Cp and 1-Cp*, stabilized by the tri-tert-butylphophinimine ligand and either C5H5 or C5Me5, were generated from the neutral dimethyl precursors and [Ph3C]+[B(C6F5)4]-. Reaction of these compounds with H2 resulted in contrasting reactions. For 1-Cp, hydrogenolysis of the Ti-CH3 group led to rapid reduction to Ti(III) and production of a cationic Ti(III) dimer, 2, presumably formed upon loss of H2 from a transiently generated Ti(IV) hydride. Compound 2 was characterized crystallographically and via its cleavage with donor solvents such as THF to form the monomeric [Cp(L)Ti(THF)2]+[B(C6F5)4]-, 3. In contrast, 1-Cp* reacted rapidly with H2 to form a cationic Ti(IV) hydride species, 4, which was resistant to reduction. While only moderately stable in solution under H2, a stable, isolable THF adduct preciptitated upon addition of THF, giving 4.THF, which was fully characterized, including via X-ray crystallography. Naked hydride 4 was very reactive toward haloarene solvents such as bromobenzene, giving the cationic bromide [Cp*(L)TiBr]+[B(C6F5)4]-, 5, which was fully characterized as its THF adduct 5.THF. The contrasting behavior of 1-Cp and 1-Cp* is a result of the greater steric protection and electron donation provided by the Cp* ligand relative to the Cp donor. PMID- 15125651 TI - Multistep electron transfer processes on dye co-sensitized nanocrystalline TiO2 films. AB - We report a method for achieving multilayer co-sensitization of nanocrystalline TiO2 films. The method is based upon an aluminum isopropoxide treatment of the monosensitized film prior to deposition of a second sensitizer. Appropriate selection of sensitizer dyes allows vectorial, multistep, electron transfer processes, resulting in a suppression of interfacial charge recombination and a significantly improved photovoltaic device performance relative to single-layer co-sensitization devices. PMID- 15125652 TI - Organocatalytic enantioselective conjugate addition to alkynones. AB - The first catalytic enantioselective addition of beta-dicarbonyl compounds to alkynones is presented. The conjugate addition proceeds in very high yields, giving a mixture of (Z)- and (E)-enones with up to 95% ee using the cinchona alkaloid [DHQ]2PHAL (5 mol %) as the catalyst. This organocatalytic enantioselective reaction has been further developed to a one-pot procedure to give the optically active (E)-enone adduct using first [DHQ]2PHAL (5 mol %), followed by Bu3P (10 mol %), as the catalysts. PMID- 15125653 TI - Layer-by-layer nanotube template synthesis. AB - Electroless deposition of gold on the pore walls of polycarbonate templates is currently the best known method for controlling inside diameters of template synthesized nanotubes. It would be very useful to have alternative template-based synthetic chemistries that yield nanotubes composed of other materials, but which still allow for precise control over the nanotube wall thickness and i.d. A film formation process that is based on layer-by-layer deposition of the film-forming material along the pore walls of the template membrane provides this desired alternative synthetic chemistry. We describe here the use of Mallouk's alpha,omega-diorganophosphonate/Zr layer-by-layer film-forming method for preparing nanotubes within the pores of alumina template membranes. We have found that this method allows accurate, quantitative, and predictable control over the wall thickness, and thus i.d., of the layered nanotubes obtained. PMID- 15125654 TI - Convenient synthesis of alkylhydrazides by the cobalt-catalyzed hydrohydrazination reaction of olefins and azodicarboxylates. AB - Treatment of olefins 2 with 1.5 equiv of di-tert-butyl azodicarboxylate (3), 1 equiv of PhSiH3, and 1.5-5 mol % of the simple Co(III) catalyst 1 in ethanol at 23 degrees C affords the Markovnikov hydrazide product for a broad range of olefins in 62-94% yield. PMID- 15125655 TI - Genome mining to identify new plant triterpenoids. AB - Arabidopsis thaliana is a well-established model organism for plant genetics, and its recently sequenced genome reveals a wealth of enzymes similar to known examples that biosynthesize secondary metabolites. We describe experiments that exploit this genomic information to identify novel terpenoids. A predicted open reading frame with high similarity to known oxidosqualene cyclases was shown to convert 3(S)-oxidosqualene to the previously unknown triterpene alcohol (3S,13S,14R)-malabarica-8,17,21-trien-3-ol, which we named thalianol. Genome mining offers a systematic approach to exhaustively characterize the biosynthetic potential of an organism, and is considerably more sensitive than classical approaches. Because even rare transcripts can be heterologously expressed at high levels, genome mining coupled to heterologous expression may be more sensitive than classical extraction approaches for isolating and characterizing trace metabolites. PMID- 15125656 TI - Copper-catalyzed electrophilic amination of diorganozinc reagents. AB - The copper-catalyzed electrophilic amination of diorganozinc reagents employing O acyl N,N-dialkyl hydroxylamine derivatives as aminating agents is described. This reaction offers a general method for the preparation of tertiary amines in high yields and is noteworthy for its convenience both in terms of reaction conditions employed (room temperature, 50 times more reactive than samples prepared by Au or Au2 deposition PMID- 15125658 TI - Electroconductive Langmuir-Blodgett films containing a carotenoid amphiphile for sugar recognition. AB - Fabrication of electroconductive molecules has attracted the most interest in nanotechnology. To date, molecules bearing a long conjugated bond have been synthesized, and they exhibited electric conduction. If molecular recognition sites could be attached to the conducting molecules, the conducting molecules can be connected with each other or wired to the desired joints, and further be applied to the chemical sensors or the nanoscale electric circuits. In this study, we synthesized a carotenoid amphiphile, which bears both the conducting part, a carotenoid moiety, and the molecular recognition site, a boronic acid moiety for specific binding of sugars. The carotenoid amphiphile was mixed with a polymerizable amphiphile, spread at the air-water interface to form a monolayer, and then polymerized by light. The resulting polymerized monolayer was deposited on gold electrodes. The electric conduction and the molecular recognition ability of the monolayer on the electrode were studied by cyclic voltammetry. The electrode responded preferentially to the redox-active sugar derivative in the solution. The electric conduction was also microscopically confirmed by the AFM current imaging technique. This manuscript has demonstrated a key step for designing molecular electronic devices based on molecular assembling, by which molecules are orientated spontaneously. PMID- 15125659 TI - Enantioselective allylation of ketone-derived benzoylhydrazones: practical synthesis of tertiary carbinamines. AB - A highly practical method for the enantioselective allylation of ketone-derived benzoylhydrazones has been developed. The previously reported strained silacycle reagent 1 reacts with a wide variety of benzoylhydrazones to give the hydrazide products with good to excellent enantioselectivity (84-97% ee). A mechanism in which the acylhydrazone becomes caovalently attached to the silane has been established. PMID- 15125660 TI - N1...N3 hydrogen bonds of A:U base pairs of RNA are stronger than those of A:T base pairs of DNA. AB - Trans-hydrogen-bond deuterium isotope effects of Watson-Crick A:U and A:T base pairs of 10 homologous RNA and DNA duplexes are compared. The isotope effect at 13C2 of adenosine residues due to deuterium/protium substitution at the imino H3 site, 2hDelta13C2, is larger in RNA than in DNA. The virtually consistent larger isotope effects in RNA suggest that the N1...N3 hydrogen bonds of A:U base pairs of RNA are stronger than those of the A:T base pairs of DNA. PMID- 15125661 TI - Inhibitors of ADP-ribosylating bacterial toxins based on oxacarbenium ion character at their transition states. AB - The bacterial exotoxins, cholera toxin (CT), pertussis toxin (PT), and diphtheria toxin (DT), interfere with specific host proteins to cause tissue damage for their respective infections. The common toxic mechanism for these agents is mono ADP-ribosylation of specific amino acids in G(s)(alpha), G(i)(alpha), and eEF-2 proteins, respectively, by the catalytic A chains of the toxins (CTA, PTA, and DTA). In the absence of acceptor proteins, these toxins also act as NAD(+)-N ribosyl hydrolases. The transition-state structures for NAD(+) hydrolysis and ADP ribosylation reactions have oxacarbenium ion character in the ribose. We designed and synthesized analogues of NAD(+) to resemble their oxacarbenium ion transition states. Inhibitors with oxacarbenium mimics replacing the NMN-ribosyl group of NAD(+) show 200-620-fold increased affinity in the hydrolytic and N-ribosyl transferase reactions catalyzed by CTA. These analogues are also inhibitors for the hydrolysis of NAD(+) by PTA with K(i) values of 24-40 microM, but bind with similar affinity to the NAD(+) substrates. Inhibition of the NAD(+) hydrolysis and ADP-ribosyl transferase reactions of DTA gave K(i) values from 19 to 48 microM. Catalytic rate enhancements by the bacterial exotoxins are small, and thus transition-state analogues cannot capture large energies of activation. In the cases of DTA and PTA, analogues known to resemble the transition states bind with approximately the same affinity as substrates. Transition-state analogue interrogation of the bacterial toxins indicates that CTA gains catalytic efficiency from modest transition-state stabilization, but DTA and PTA catalyze ADP-ribosyl transferase reactions more from ground-state destabilization. pH dependence of inhibitor action indicated that both neutral and cationic forms of transition-state analogues bind to DTA with similar affinity. The origin of this similarity is proposed to reside in the cationic nature of NAD(+) both as substrate and at the transition state. PMID- 15125662 TI - Oxidative fragmentation of hydroxy octadecadienoates generates biologically active gamma-hydroxyalkenals. AB - Oxidative fragmentation of polyunsaturated fatty acids (PUFAs) in vivo generates cytotoxic aldehydes. Among these, 4-hydroxynon-2-enal and analogous gamma hydroxyalkenal phosphatidylcholines (PCs) have attracted attention because these oxidatively truncated lipids are biologically active and have been implicated in diseases. A previous study showed that hydroxydienes, generated by allylic oxygenation of linoleic acid, are unreactive toward oxidative fragmentation. We now show that, in the presence of hydroperoxides, hydroxydienes fragment as readily as the corresponding hydroperoxydienes, generating gamma-hydroxyalkenals. In a physiomimetic model study, myeloperoxidase-promoted free radical-induced fragmentation of either hydroperoxy- or hydroxyoctecadienoate esters of 2-lyso-PC in small unilamellar vesicles produced the 9-hydroxy-12-oxododec-10-enoic acid (HODA) ester HODA-PC. Therefore, hydroxydienes, that are generally more abundant in vivo than hydroperoxydienes, are plausible intermediates in the production of oxidatively truncated lipids in vivo where a constant flux of radicals and hydroperoxides is present. Our findings also show that the formation of dioxetane intermediates through peroxyradical cyclization is not required to achieve oxidative fragmentation of PUFAs. PMID- 15125663 TI - Probing the glycosidic linkage: UV and IR ion-dip spectroscopy of a lactoside. AB - The beta(1-->4) glycosidic linkage found in lactose is a prevalent structural motif in many carbohydrates and glycoconjugates. Using UV and IR ion-dip spectroscopies to probe benzyl lactoside isolated in the gas phase, we find that the disaccharide unit adopts only a single, rigid structure. Its fully resolved infrared ion-dip spectrum is in excellent agreement with that of the global minimum structure computed ab initio. This has glycosidic torsion angles of phi(H) (H1-C1-O-C4') approximately 180 degrees and psi(H) (C1-O-C4'-H4') approximately 0 degrees which correspond to a rotation of approximately 150 degrees about the glycosidic bond compared to the accepted solution-phase conformation. We discuss the biological implications of this discovery and the generality of the strategies employed in making it. PMID- 15125664 TI - Activation of crystalline cellulose surfaces through the chemoenzymatic modification of xyloglucan. AB - Cellulose constitutes an important raw material for many industries. However, the superb load-bearing properties of cellulose are accompanied by poor chemical reactivity. The hydroxyl groups on cellulose surfaces can be reacted but usually not without loss of fiber integrity and strength. Here, we describe a novel chemoenzymatic approach for the efficient incorporation of chemical functionality onto cellulose surfaces. The modification is brought about by using a transglycosylating enzyme, xyloglucan endotranglycosylase, to join chemically modified xyloglucan oligosaccharides to xyloglucan, which has a naturally high affinity to cellulose. Binding of the chemically modified hemicellulose molecules can thus be used to attach a wide variety of chemical moieties without disruption of the individual fiber or fiber matrix. PMID- 15125665 TI - A PELDOR-based nanometer distance ruler for oligonucleotides. AB - A pulsed electron paramagnetic resonance (EPR) spectroscopic ruler for oligonucleotides was developed using a series of duplex DNAs. The spin-labeling is accomplished during solid-phase synthesis of the oligonucleotides utilizing a palladium-catalyzed cross-coupling reaction between 5-iodo-2'-deoxyuridine and the rigid spin-label 2,2,5,5-tetramethyl-pyrrolin-1-yloxyl-3-acetylene (TPA). 4 Pulse electron double resonance (PELDOR) was then used to measure the intramolecular spin-spin distances via the dipolar coupling, yielding spin-spin distances of 19.2, 23.3, 34.7, 44.8, and 52.5 A. Employing a full-atom force field with explicit water, molecular dynamic (MD) simulations on the same spin labeled oligonucleotides in their duplex B-form gave spin-spin distances of 19.6, 21.4, 33.0, 43.3, and 52.5 A, respectively, in very good agreement with the measured distances. This shows that the oligonucleotides adopt a B-form duplex structure also in frozen aqueous buffer solution. It also demonstrates that the combined use of site-directed spin-labeling, PELDOR experiments, and MD simulations can yield a microscopic picture about the overall structure of oligonucleotides. The technique is also applicable to more complex systems, like ribozymes or DNA/RNA-protein complexes, which are difficult to access by NMR or X ray crystallography. PMID- 15125666 TI - Template assembled cyclopeptides as multimeric system for integrin targeting and endocytosis. AB - The alpha(V)beta(3) integrin receptor plays an important role in human metastasis and tumor-induced angiogenesis. c[-RGDfV-] peptide represents a selective alpha(V)beta(3) integrin ligand that has been extensively used for research, therapy, and diagnosis of neoangiogenesis. We report here the modular synthesis and biological characterization of template assembled cyclopeptides as a multimeric system for targeting and endocytosis of cells expressing alpha(V)beta(3) integrin. c[-RGDfK-] was cleanly assembled in a multivalent mode by chemoselective oxime bond formation to a cyclodecapeptides template labeled by different reporter groups. Binding propensity to the alpha(V)beta(3) receptor and the associated good uptake property displayed by the multivalent molecules demonstrated the interest in the RAFT molecule to design new multimeric system with hitherto unreported properties. These compounds offer an interesting perspective for the reevaluation of integrins as angiogenesis regulators (Hynes, R. O. Nature Med. 2003, 9, 918-921) as well as for the design of more sophisticated systems such as molecular conjugate vectors. PMID- 15125667 TI - Encoding method for OBOC small molecule libraries using a biphasic approach for ladder-synthesis of coding tags. AB - In the "one-bead one-compound" (OBOC) combinatorial library method, each compound bead displays only one compound entity. Hundreds of thousands to millions of compound beads can be synthesized rapidly and screened simultaneously. Positive compound beads are then isolated for structural analysis. To fully exploit the power of OBOC combinatorial small molecule libraries, a robust and high throughput encoding method is needed to decode the positive compound beads. In this paper, we report on the development of a novel encoding strategy that combines the concepts of ladder-synthesis and chemical encoding on bilayer beads. In these encoded libraries, small molecule compounds are displayed on the bead surface, and cleavable coding tags consisting of a series of truncated molecules reside in the bead interior. Such a library can be easily constructed using the biphasic approach (J. Am. Chem. Soc.2002, 124, 7678) to topologically segregate the functionalities of the beads during library synthesis. The ladder members and coding tags are then released for MALDI-TOF-MS analysis. To simplify the interpretation of the mass spectra, we purposely add bromine into the cleavable linker so that the cleavage products generate a characteristic isotope fingerprint. The chemical structure of library compounds can be determined by analyzing the mass differences between adjacent peaks on the mass spectra. This encoding strategy also provides valuable information on the quality of the testing compound on the surface of the bead. To validate this methodology, a model OBOC small molecule library with 12,288 members was synthesized on TentaGel beads and screened against streptavidin. The chemical structures of the compound on each positive bead were unambiguously identified. PMID- 15125668 TI - Evidence for specific solvation of two halocarbene amides. AB - Laser flash photolysis (LFP, 308 nm) of endo-10-halo-10'-N,N dimethylcarboxamidetricyclo[4.3.1.0]-deca-2,4-diene (1Cl and 1F) releases indan and halocarbene amide (2Cl and 2F). Although the carbenes are not UV-vis active, they react rapidly with pyridine to form ylides (4Cl, 4F), which are readily detected in LFP experiments (lambda(max) = 450 nm). Dioxane decreases the observed rate of carbene reaction with pyridine in CF(2)ClCFCl(2). Small amounts of THF decrease the observed rate of reaction of carbene 2F with pyridine but increase the rate of reaction of carbene 2Cl with pyridine. LFP (266 nm) of dienes 1Cl and 1F in CF(2)ClCFCl(2) with IR detection produces carbenes 2Cl and 2F with carbonyl vibrations at 1635 and 1650 cm(-1), respectively. In dioxane or THF solvent, LFP produces the corresponding ether ylides (5Cl, 5F) by capture of carbenes 2Cl and 2F. The ylides have broad carbonyl vibrations between 1560 and 1610 cm(-1). The addition of a small amount of dioxane in CFCl(2)CF(2)Cl extends the lifetime of the carbene. This observation, together with the ether-induced retardation of the rates of carbene capture by tetramethylethylene and pyridine, is evidence for solvation of the carbene by dioxane. PMID- 15125669 TI - Proton-coupled electron transfer in soybean lipoxygenase. AB - The proton-coupled electron transfer reaction catalyzed by soybean lipoxygenase-1 is studied with a multistate continuum theory that represents the transferring hydrogen nucleus as a quantum mechanical wave function. The inner-sphere reorganization energy of the iron cofactor is calculated with density functional theory, and the outer-sphere reorganization energy of the protein is calculated with the frequency-resolved cavity model for conformations obtained with docking simulations. Both classical and quantum mechanical treatments of the proton donor acceptor vibrational motion are presented. The temperature dependence of the calculated rates and kinetic isotope effects is in agreement with the experimental data. The weak temperature dependence of the rates is due to the relatively small free energy barrier arising from a balance between the reorganization energy and the reaction free energy. The unusually high deuterium kinetic isotope effect of 81 is due to the small overlap of the reactant and product proton vibrational wave functions and the dominance of the lowest energy reactant and product vibronic states in the tunneling process. The temperature dependence of the kinetic isotope effect is strongly influenced by the proton donor-acceptor distance with the dominant contribution to the overall rate. This dominant proton donor-acceptor distance is significantly smaller than the equilibrium donor-acceptor distance and is determined by a balance between the larger coupling and the smaller Boltzmann probability as the distance decreases. Thus, the proton donor-acceptor vibrational motion plays a vital role in decreasing the dominant donor-acceptor distance relative to its equilibrium value to facilitate the proton-coupled electron transfer reaction. PMID- 15125670 TI - New synthesis and insight into the structure of blue ultramarine pigments. AB - A new and easy method for preparing blue sodalite pigments which involves high temperature calcination of sodalite samples synthesized with aluminum sulfate and an organic template, is presented. Calcination generated the S(3)(-) and S(2)(-) radicals, and the effects of the Al/Si ratio and the calcination temperature on the nature and amounts of the radicals were examined. The radicals were characterized in detail by continuous wave and pulsed EPR at X- and W-band frequencies (approximately 9 and 95 GHz, respectively) complemented by UV-vis measurements. The high-field electron-paramagnetic resonance (EPR) measurements allowed us to clearly resolve the g anisotropy of S(3)(-) and W-band electron nuclear double resonance (ENDOR) measurements detected strong coupling with extra framework (23)Na cations and weak coupling with framework (27)Al. On the basis of the spectroscopic results and density functional theory (DFT) calculations of the g-tensors of S(3)(-) and S(2)(-) radicals, the EPR signals were attributed to three different radicals, all with the open structure C(2v), that are located within the sodalite beta cages. While two of these radicals are well isolated, the third one is associated with an exchange-narrowed signal originating from S(3)(-) radicals in nearby sodalite cages. PMID- 15125671 TI - Do spin state changes matter in organometallic chemistry? A computational study. AB - Spin changes occur often in organometallic chemistry, and their effect on kinetics is not well understood. We report computations on the singlet and triplet potential energy surfaces of several processes of this type and show that the topology of the individual surfaces, as well as of the crossing regions between them, can be used to rationalize the observed reactivity in all cases. In particular, the slow addition of dihydrogen to W[N(CH(2)CH(2)NSiMe(3))(3)]H (Schrock, R. R.; Shih, K. Y.; Dobbs, D. A.; Davis, W. M. J. Am. Chem. Soc. 1995, 117, 6609) is shown to be a "spin-blocked" reaction with a high barrier due to the crossing between reactant triplet and product singlet surfaces. In contrast, addition of CO to TpCo(CO) (Detrich, J. L.; Reinaud, O. M.; Rheingold, A. L.; Theopold, K. H. J. Am. Chem. Soc. 1995, 117, 11745) is fast because the triplet and singlet surfaces cross at low energy. Particular care is taken to use DFT methods which are in adequate agreement with experimental and high-level computational energetics for these systems. PMID- 15125672 TI - Synthesis of polyphenylene dendrimers related to "cubic graphite". AB - Four large, 6-fold symmetric, polyphenylene hydrocarbons have been prepared by short syntheses that chiefly employed alkyne trimerization, palladium-catalyzed coupling, and Diels-Alder reactions. The two largest of these molecules, hexakis[4'-(pentaphenylphenyl)biphenyl-4-yl]benzene (4, C(294)H(198)) and hexakis[4'-(2,3,4,5-tetraphenylphenyl)biphenyl-4-yl]benzene (5, C(258)H(174)) are substructures of "phenylogous cubic graphite", and the other two, hexakis(2',3',4',5',6'-pentaphenylbiphenyl-4-yl)benzene (26, C(258)H(174)) and hexakis(2',3',4',5'-tetraphenylbiphenyl-4-yl)benzene (25, C(222)H(150)) are strongly pitched, six-bladed molecular propellers. The X-ray crystal structure of compound 26 has also been determined; dendrimer 26 is at present the largest crystallographically characterized hydrocarbon. PMID- 15125673 TI - Mechanism of host-guest complexation by cucurbituril. AB - The factors affecting host-guest complexation between the molecular container compound cucurbit[6]uril (CB6) and various guests in aqueous solution are studied, and a detailed complexation mechanism in the presence of cations is derived. The formation of the supramolecular complex is studied in detail for cyclohexylmethylammonium ion as guest. The kinetics and thermodynamics of complexation is monitored by NMR as a function of temperature, salt concentration, and cation size. The binding constants and the ingression rate constants decrease with increasing salt concentration and cation-binding constant, in agreement with a competitive binding of the ammonium site of the guest and the metal cation with the ureido carbonyl portals of CB6. Studies as a function of guest size indicate that the effective container volume of the CB6 cavity is approximately 105 A(3). It is suggested that larger guests are excluded for two reasons: a high activation barrier for ingression imposed by the tight CB6 portals and a destabilization of the complex due to steric repulsion inside. For example, in the case of the nearly spherical azoalkane homologues 2,3 diazabicyclo[2.2.1]hept-2-ene (DBH, volume ca. 96 A(3)) and 2,3 diazabicyclo[2.2.2]oct-2-ene (DBO, volume ca. 110 A(3)), the former forms the CB6 complex promptly with a sizable binding constant (1300 M(-1)), while the latter does not form a complex even after several months at optimized complexation conditions. Molecular mechanics calculations are performed for several CB6/guest complexes. A qualitative agreement is found between experimental and calculated activation energies for ingression as a function of both guest size and state of protonation. The potential role of constrictive binding by CB6 is discussed. PMID- 15125674 TI - Synthesis, structure solution, characterization, and catalytic properties of TNU 10: a high-silica zeolite with the STI topology. AB - A high-silica zeolite (Si/Al = 7.1) with the STI framework topology, denoted TNU 10, has been synthesized in the presence of 1,4-bis(N-methylpyrrolidinium)butane and Na(+) cations as structure-directing agents, and its structure in the proton form has been refined against laboratory powder X-ray data in space group Fmmm (a = 13.533(1) A, b = 17.925(2) A, c = 17.651(2) A). The space group symmetry is supported by electron diffraction and energy minimization studies. The as-made and proton form of TNU-10 are extensively characterized by elemental and thermal analyses, scanning electron microscopy, N(2) adsorption, multinuclear solid-state NMR, IR, and temperature-programmed desorption of ammonia, and the location of the organic structure-directing agent in the channel system is determined by molecular modeling. The catalytic properties of H-TNU-10 and Co-TNU-10 are evaluated for the skeletal isomerization of 1-butene to isobutene and the selective reduction of NO with methane, respectively. When compared to H ferrierite, a low selectivity to isobutene is observed for H-TNU-10. However, it is found that Co-TNU-10 exhibits a maximum NO conversion of 93% at 823 K under conditions of high concentrations of methane (16,000 ppm) and water vapor (10%) and in the presence of 2.6% O(2), which is considerable higher than even the value (74%) obtained from Co-ferrierite, known as the best catalyst for this reaction, under the identical conditions. PMID- 15125675 TI - A mechanistic investigation of the polymerization of ethylene catalyzed by neutral Ni(II) complexes derived from bulky anilinotropone ligands. AB - An extensive mechanistic investigation has been carried out on ethylene polymerizations catalyzed by neutral Ni(II) catalysts derived from bulky anilinotropone ligands. Complexes and precatalysts prepared include aryl derivatives [(2,6-i-Pr(2)C(6)H(3))NC(7)H(4)O(7-Aryl)Ni(Ph)(PPh(3))] (9, Aryl = phenyl(a), 1-naphthyl(b), p-methoxyphenyl(c), p-trifluoromethylphenyl(d)), alkyl derivatives [[(2,6-(i)Pr(2)C(6)H(3))NC(7)H(5)O]Ni(R)(2,4-lutidine)] (16, R = Et (a), n-Pr (b)) and [[(2,6-(i)Pr(2)C(6)H(3))NC(7)H(5)O]Ni(R)(PPh(3))] (17, R = Et (a), n-Pr (b), n-hexyl (c), i-Pr (d)), and the nickel hydride complex [[(2,6 (i)Pr(2)C(6)H(3))NC(7)H(5)O]Ni(H)(PPh(3))], 20. Branched polyethylenes are produced at 40-80 degrees C in toluene with M(n) values in the 100-200K range and molecular weight distributions of ca. 1.4-2.2. Branching ranges from 15 to 64 branches/1000 carbons depending on temperature and ethylene pressure. The electron-withdrawing -CF(3) substituent on the 7-aryl group increases activity but has little effect on branching and molecular weight. NMR experiments establish that in the case of the PPh(3)-substituted systems, the catalyst rests as an equilibrating mixture of the alkyl phosphine and the alkyl ethylene complexes. At high ethylene pressures, the turnover frequency saturates, indicating that the equilibrium has shifted nearly completely to the alkyl olefin complex. Under these conditions, the barriers to migratory insertion were determined to be ca. 16-17 kcal/mol for 9a, 9c, 9d, and 16a. Extraction of 2,6 lutidine from complexes 16a,b yields highly dynamic beta-agostic alkyl complexes [[(2,6-i-Pr(2)C(6)H(3))NC(7)H(5)O]Ni(Et)] 21 and [[(2,6-i Pr(2)C(6)H(3))NC(7)H(5)O]Ni(i-Pr)] 22. Free energy barriers to nickel-carbon bond rotation and beta-hydride elimination of 11.1 and ca. 17 kcal/mol, respectively, were determined for 22. Themolysis of 17c at 50 degrees C generates hydride 20 and hexene and occurs by two pathways, one independent of [PPh(3)] and one retarded by PPh(3). At much slower rates, hydride 20 reductively eliminates free ligand, which ultimately generates a bis-ligand complex, 25. Catalyst decay under polymerization conditions was shown to occur by a similar process to generate free ligand and a bis-ligand complex formed by reaction of free ligand with an active catalyst species. The major chain transfer route is a simple beta elimination process, not chain transfer to monomer. PMID- 15125676 TI - Vibrational coupling, isotopic editing, and beta-sheet structure in a membrane bound polypeptide. AB - The N-acetylated hexapeptide WLLLLL (AcWL5) partitions into lipid membranes and is believed to assemble into an antiparallel beta-sheet. As a test of this structural assignment, the peptide bonds of residues 2-6 were labeled with (13)C and allowed to adsorb onto a supported lipid membrane. Peptides bound to the membrane were examined for evidence of coupling between the labeled vibrational modes in adjacent beta-strands with internal reflection infrared spectroscopy. Experimental results indicate that the amide I absorption band in D(2)O (i.e., amide I') attributable to (13)C is selectively enhanced when the label is at any one of several positions along the peptide backbone. Simulations employing an excitonic model with through-bond and through-space interactions were performed on AcWL5 models in parallel and antiparallel beta-sheet configurations. The simulations yield spectra in good agreement with the experimental results, accounting for the enhancement of both (13)C band intensities and band frequencies. They also yield insight into the physical origin and structure selectivity of the distinctive amide I' band shapes that arise in isotopically edited spectra. It is concluded that the beta-sheet formed by membrane-bound AcWL5 is indeed antiparallel, and the enhancement of (13)C bands in the infrared spectra of these peptides is caused by both interstrand and intrastrand coupling to (12)C modes. PMID- 15125677 TI - Electronic band structure of titania semiconductor nanosheets revealed by electrochemical and photoelectrochemical studies. AB - Electrochemical and photoelectrochemical studies were conducted on self-assembled multilayer films of titania nanosheets on a conductive ITO substrate. Cyclic voltammogram (CV) curves indicated that the titania nanosheet electrode underwent insertion/extraction of Li(+) ions into/from the nanosheet galleries, associated with reduction/oxidation of Ti(4+)/Ti(3+). These processes accompanied reversible changes in UV-vis absorption of the titania nanosheet electrodes. Applying a negative bias of -1.3 V (vs Ag/Ag(+)) and lower brought about absorption reduction where the wavelength is shorter than 323 nm, and vice versa, indicating a flat-band potential of (approximately) -1.3 V and a band gap energy of 3.84 eV. Photocurrents were generated from the titania nanosheet electrodes under a positive bias. The onset potential for photocurrent generation from the titania nanosheet electrodes was around -1.27 V, and the band gap energy estimated from the photocurrent action spectra was 3.82 eV, in excellent agreement with the values obtained from the spectroelectrochemical data. The lack of difference in the band gap energies for titania nanosheet electrodes with different numbers of layers suggests that a nanosheet is electronically isolated in multilayer assemblies without affecting the electronic state of neighboring nanosheets. Similar measurements on the anatase-type TiO(2) electrode revealed that the lower edge of the conduction band for the titania nanosheet is approximately 0.1 V higher than that for anatase, while the upper edge of the valence band is 0.5 V lower. PMID- 15125678 TI - X-ray magnetic circular dichroism of Pseudomonas aeruginosa nickel(II) azurin. AB - We show that X-ray magnetic circular dichroism (XMCD) can be employed to probe the oxidation states and other electronic structural features of nickel active sites in proteins. As a calibration standard, we have measured XMCD and X-ray absorption (XAS) spectra for the nickel(II) derivative of Pseudomonas aeruginosa azurin (NiAz). Our analysis of these spectra confirms that the electronic ground state of NiAz is high-spin (S = 1); we also find that the L(3)-centroid energy is 853.1(1) eV, the branching ratio is 0.722(4), and the magnetic moment is 1.9(4) mu(B). Density functional theory (DFT) calculations on model NiAz structures establish that orbitals 3d(x2-y2) and 3d(z2) are the two valence holes in the high-spin Ni(II) ground state, and in accord with the experimentally determined orbital magnetic moment, the DFT results also demonstrate that both holes are highly delocalized, with 3d(x2-y2) having much greater ligand character. PMID- 15125679 TI - WISE NMR characterization of nanoscale heterogeneity and mobility in supercontracted Nephila clavipes spider dragline silk. AB - The addition of water to spider dragline silk results in fiber contraction to 50% its initial length and significant changes to the mechanical properties of the silk. This event has been termed supercontraction. A decrease in strength and increase in elasticity have been reported when the silk is in contact with water. Two-dimensional wide-line separation (WISE) nuclear magnetic resonance (NMR) is implemented to correlate (13)C chemical shifts with mobility by observing the corresponding (1)H line widths and line shapes in water-saturated spider dragline silk. The WISE NMR spectrum of the native silk exhibits (1)H line widths that are approximately 40 kHz for all carbon environments characteristic of a rigid organic system. In contrast, the water-saturated case displays a component of the (1)H line that is narrowed to approximately 5 kHz for the glycine C(alpha) and a newly resolved alanine helical environment while the alanine C(beta) corresponding to the beta-sheet conformation remains broad. These results indicate that water permeates the amorphous, glycine-rich matrix and not the crystalline, polyalanine beta-sheets. A delay time is added to the WISE NMR pulse sequence to monitor spin diffusion between the amorphous, mobile region and the crystalline domains. The time required for spin diffusion to reach spatial equilibrium is related to the length scale of the polyalanine crystallites. This technique is employed to measure crystalline domain sizes on the nanometer length scale in water-solvated spider dragline silk. These results provide further insight into the structure of spider silk and mechanism of supercontraction. PMID- 15125680 TI - Biomolecular NMR using a microcoil NMR probe--new technique for the chemical shift assignment of aromatic side chains in proteins. AB - A specially designed microcoil probe for use in biomolecular NMR spectroscopy is presented. The microcoil probe shows a mass-based sensitivity increase of a minimal factor of 7.5, allowing for the first time routine biomolecular NMR spectroscopy with microgram amounts of proteins. In addition, the exceptional radio frequency capabilities of this probe allowed us to record an aliphatic aromatic HCCH-TOCSY spectrum for the first time. Using this spectrum, the side chains of aliphatic and aromatic amino acids can be completely assigned using only a single experiment. Using the conserved hypothetical protein TM0979 from Thermotoga maritima, we demonstrate the capabilities of this microcoil NMR probe to completely pursue the sequence specific backbone assignment with less than 500 microg of (13)C,(15)N labeled protein. PMID- 15125681 TI - Ensemble approach for NMR structure refinement against (1)H paramagnetic relaxation enhancement data arising from a flexible paramagnetic group attached to a macromolecule. AB - Paramagnetic relaxation enhancement (PRE) measurements on (1)H nuclei have the potential to play an important role in NMR structure determination of macromolecules by providing unique long-range (10-35 A) distance information. Recent methodological advances for covalently attaching paramagnetic groups at specific sites on both proteins and nucleic acids have permitted the application of the PRE to various biological macromolecules. However, because artificially introduced paramagnetic groups are exposed to solvent and linked to the macromolecule by several freely rotatable bonds, they are intrinsically flexible. This renders conventional back-calculation of the (1)H-PRE using a single-point representation inaccurate, thereby severely limiting the utility of the (1)H-PRE as a tool for structure refinement. To circumvent these limitations, we have developed a theoretical framework and computational strategy with which to accurately back-calculate (1)H-PREs arising from flexible paramagnetic groups attached to macromolecules. In this scheme, the (1)H-PRE is calculated using a modified Solomon-Bloembergen equation incorporating a "model-free" formalism, based on a multiple-structure representation of the paramagnetic group in simulated annealing calculations. The ensemble approach for (1)H-PRE back calculation was examined using several SRY/DNA complexes incorporating dT-EDTA Mn(2+) at three distinct sites in the DNA, permitting a large data set comprising 435 experimental backbone and side-chain (1)H-PREs to be obtained in a straightforward manner from 2D through-bond correlation experiments. Calculations employing complete cross-validation demonstrate that the ensemble representation provides a means to accurately utilize backbone and side-chain (1)H-PRE data arising from a flexible paramagnetic group in structure refinement. The results of (1)H-PRE based refinement, in conjunction with previously obtained NMR restraints, indicate that significant gains in accuracy can be readily obtained. This is particularly significant in the case of macromolecular complexes where intermolecular translational restraints derived from nuclear Overhauser enhancement data may be limited. PMID- 15125682 TI - Direct experimental evaluation of charge scheme performance by a molecular charge meter. AB - The remarkable advances accomplished in the past two decades in theoretical and computational capabilities have made the in silico study of complex chemical systems feasible. However, this progress is in strong contrast to the lag in experimental capabilities relating to the measurement of fundamental chemical quantities within convoluted environments such as solvents or protein milieu. As a result, many works rely extensively on predictions provided by ab initio methodologies without having independent experimental support. Such a proliferation of theory and computational approaches without being substantiated by appropriate experimental data is undesirable. The feasibility of using nickel bacteriochlorophyll as a molecular potentiometer was recently demonstrated for the systematic evaluation of fragmental charge density transfer for metal complexes in solution, thus providing an experimental assay with high accuracy and sensitivity (better than +/-0.005 e(-); Yerushalmi, R.; Baldridge, K. K.; Scherz, A. J. Am. Chem. Soc. 2003, 125, 12706-12707). Here the experimentally determined fragmental charge density transfer values measured by the molecular potentiometer for metal complexes in solvent are used to provide, for the first time, an independent and critical experimental evaluation of theoretical approaches commonly used in determining atomic charges and fragmental charge density transfer among interacting molecular systems. Importantly, these findings indicate that the natural population analysis (NPA) charge analysis is highly robust and well-suited for determining charge transfer processes involving donor acceptor coordination interactions. The majority of computational charge schemes fail to provide an accurate chemical picture for the whole range of systems considered here. In cases where the role of electronic correlation varies significantly among chemically related structures, as with mono- and biligated complexes, the widely used electrostatic potential fit-based methods for evaluating atomic charges may prove to be problematic for predictive studies. In such cases, alternative methods that do not rely on the net dipole moment or other higher multipoles of the system for determining charges should be employed. PMID- 15125683 TI - Gas-phase hydrogen/deuterium exchange and conformations of deprotonated flavonoids and gas-phase acidities of flavonoids. AB - Gas-phase hydrogen/deuterium (H/D) exchange was used to probe the conformations, gas-phase acidities, and sites of deprotonation of isomeric flavonoid aglycons and glycosides. The flavonoids in each isomer series were differentiated on the basis of their relative rate constants and total numbers of exchanges. For example, flavonoids that possess neohesperidose-type disaccharides may undergo faster and far more extensive exchange than isomeric rutinoside flavonoids. The structural factors that promote or prevent H/D exchange were identified and correlated with collisionally activated dissociation (CAD) patterns and/or molecular modeling data (both high-level ab initio acidity calculations and conformational analysis with molecular dynamics (MD) simulations), thus providing a framework for the use of H/D exchange reactions in the structural elucidation of new flavonoids. PMID- 15125684 TI - Surface adsorption and transfer of organomercaptans to colloidal gold and direct identification by matrix assisted laser desorption/ionization mass spectrometry. AB - The adsorption characteristics of two different organomercaptan adsorbates, 5-((2 (and-3)-S-(acetylmercapto)succinoyl)amino)fluorescein (SAMSA) and the peptide Cys Lys-Trp-Ala-Lys-Trp-Ala-Trp (CKWAKWAK), on colloidal Au were studied, and the conjugates produced were characterized by UV-vis spectroscopy, transmission electron microscopy, and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Fluorescence difference measurements of free thiols in solution were used to assemble surface adsorption isotherms on Au colloid revealing surface coverages of 1.0 x 10(14) molecules cm(-2) for SAMSA and 3.1 x 10(14) molecules cm(-2) for CKWAKWAK. The free energies of adsorption were calculated to be -48.4 kJ/mol for SAMSA and -49.2 kJ/mol for CKWAKWAK. UV-visible absorption spectroscopy and transmission electron microscopy reveal that the thiol/colloid conjugates flocculate under conditions where the net charge per colloid is small or neutral and that flocculated colloids can be resuspended by a change in pH to more basic for the acidic SAMSA/Au conjugates or to more acidic for the basic CKWAKWAK/Au conjugates. The reversible flocculation allows the conjugates to be readily separated from free adsorbate in solution and thereby prepared for further characterization. CKWAKWAK/colloid conjugates were analyzed by MALDI-MS, and the mass spectra show (M + H)(+), (M + Na)(+), and (M + K)(+) ions attributable to the peptide. The manipulations studied here constitute a powerful complement to microfluidic-based separation and analysis methods. Conjugating mass-limited analytes to Au colloids makes it possible to sequester and transfer small quantities of analytes with high efficiency. PMID- 15125685 TI - A novel role of Mgm1p, a dynamin-related GTPase, in ATP synthase assembly and cristae formation/maintenance. AB - In Saccharomyces cerevisiae, two mitochondrial inner-membrane proteins play critical roles in organellar morphology. One is a dynamin-related GTPase, Mgm1p, which participates in mitochondrial fusion. Another is Tim11p, which is required for oligomeric assembly of F1Fo-ATP synthase, which generates ATP through oxidative phosphorylation. Our data bring these findings together and define a novel role for Mgm1p in the formation and maintenance of mitochondrial cristae. We show that Mgm1p serves as an upstream regulator of Tim11p protein stability, ATP synthase assembly, cristae morphology and cytochrome c storage within cristae. PMID- 15125686 TI - Copper pathways in Plasmodium falciparum infected erythrocytes indicate an efflux role for the copper P-ATPase. AB - Copper, like iron, is a transition metal that can generate oxygen radicals by the Fenton reaction. The Plasmodium parasite invades an erythrocyte host cell containing 20 microM copper, of which 70% is contained in the Cu/Zn SOD (cuprozinc superoxide dismutase). In the present study, we follow the copper pathways in the Plasmodium-infected erythrocyte. Metal-determination analysis shows that the total copper content of Percoll-purified trophozoite-stage infected erythrocytes is 66% that of uninfected erythrocytes. This decrease parallels the decrease seen in Cu/Zn SOD levels in parasite-infected erythrocytes. Neocuproine, an intracellular copper chelator, arrests parasites at the ring-to-trophozoite stage transition and also specifically decreases intraparasitic levels of Cu/Zn SOD and catalase. Up to 150 microM BCS (2,9 dimethyl-4,7-diphenyl-1,10-phenanthrolinedisulphonic acid), an extracellular copper chelator, has no effect on parasite growth. We characterized a single copy PfCuP-ATPase (Plasmodium falciparum copper P-ATPase) transporter, which, like the Crypto-sporidium parvum copper P-ATPase, has a single copper-binding domain: 'Met Xaa-Cys-Xaa-Xaa-Cys'. Recombinant expression of the N-terminal metal-binding domain reveals that the protein specifically binds reduced copper. Transcription of the PfCuP-ATPase gene is the highest at late ring stage/early trophozoite, and is down-regulated in the presence of neocuproine. Immunofluorescence and electron microscopy indicate the transporter to be both in the parasite and on the erythrocyte membrane. Both the decrease in total copper and the location of the PfCuP-ATPase gene indicate a copper-efflux pathway from the infected erythrocyte. PMID- 15125687 TI - Kinetic and structural analysis of the increased affinity of enoyl-ACP (acyl carrier protein) reductase for triclosan in the presence of NAD+. AB - The binding of enoyl-ACP (acyl-carrier protein) reductase from Plasmodium falciparum (PfENR) with its substrates and inhibitors has been analysed by SPR (surface plasmon resonance). The binding of the substrate analogue crotonoyl-CoA and coenzyme NADH to PfENR was monitored in real time by observing changes in response units. The binding constants determined for crotonoyl-CoA and NADH were 1.6x10(4) M(-1) and 1.9x10(4) M(-1) respectively. Triclosan, which has recently been demonstrated as a potent antimalarial agent, bound to the enzyme with a binding constant of 1.08x10(5) M(-1). However, there was a 300-fold increase in the binding constant in the presence of NAD+. The increase in the binding constant was due to a 17 times increase in the association rate constant (k(1)) from 741 M(-1) x s(-1) to 1.3x10(4) M(-1) x s(-1) and a 16 times decrease in the dissociation rate constant (k(-1)) from 6.84x10(-3) s(-1) to 4.2x10(-4) s(-1). These values are in agreement with those determined by steady-state kinetic analysis of the inhibition reaction [Kapoor, Reddy, Krishnasastry, N. Surolia and A. Surolia (2004) Biochem. J. 381, 719-724]. In SPR experiments, the binding of NAD+ to PfENR was not detected. However, a binding constant of 6.5x10(4) M(-1) was obtained in the presence of triclosan. Further support for these observations was provided by the crystal structures of the binary and ternary complexes of PfENR. Thus the dramatic enhancement in the binding affinity of both triclosan and NAD+ in the ternary complex can be explained by increased van der Waals contacts in the ternary complex, facilitated by the movement of residues 318-324 of the substrate-binding loop and the nicotinamide ring of NAD+. Interestingly, the results of the present study also provide a rationale for the increased affinity of NAD+ for the enzyme in the ternary complex. PMID- 15125688 TI - Rebound acid hypersecretion. PMID- 15125689 TI - Rebound hypersecretion after inhibition of gastric acid secretion. AB - Drugs inhibiting gastric acid secretion are widely used because of the high prevalence of acid-related disorders. However, from clinical experience it seems that symptom relapse is common after withdrawal of these drugs. Experimental as well as clinical studies have demonstrated an increased acid secretion after a period of treatment with either histamine 2 receptor antagonists or proton pump inhibitors. Rebound hypersecretion is likely to reflect the following sequence of events: Long-term inhibition of acid output is accompanied by elevated serum gastrin levels, leading to enterochromaffin-like cell activation and proliferation, resulting in increased amounts of histamine being mobilized from these cells to stimulate the parietal cells. The clinical consequences of rebound hypersecretion have not been settled. PMID- 15125690 TI - Acidification alters antiarrhythmic drug blockade of the ether-a-go-go-related Gene (HERG) Channels. AB - Acidosis is one of the important deleterious factors during myocardial ischaemia and reperfusion. The ether-a-go-go-related gene, HERG, is a primary target for blockade by many drugs including dofetilide, quinidine and azimilide. While most drugs lose their efficacy against arrhythmias associated with myocardial ischaemia and reperfusion, dofetilide remains effective. The unique ability of dofetilide to terminate ischaemia-induced arrhythmias is not yet fully explained. The aim of the present study is to elucidate the acidification modulation of antiarrhythmic drugs blockade of HERG channels. The human gene HERG encoding K+ channels were expressed in Xenopus oocytes, and Whole-cell macroscopic currents of Xenopus oocytes were recorded with conventional two-electrode techniques. The inhibitory effects of dofetilide (0.25 microM) were significantly enhanced with decreasing pH (from 7.5 to 6.5). The percent block of dofetilide under pH 6.5 at 0 mV was 69+/-6.1% versus 54+/-3.0% under pH 7.5 (n=7, P<0.05). The IC50 values, determined by the Hill equation with the currents recorded at 0 mV, were decreased by approximately half from 192+/-23 nM with pH 7.5 to 93+/-15 nM with pH 6.5 (P<0.01). Acidification weakened the inhibitory effects of quinidine and azimilide on HERG channels. At 0 mV, the percent block of quinidine (10 microM) under pH 6.5 was 24+/-2.8% versus 62.5+/-9.0% under pH 7.5 (n=4, P<0.01), The percent block of azimilide (10 microM) under pH 6.5 was similar to that under pH 7.5 (n=6). Acidification markedly potentiated dofetilide blockade of the HERG channels but weakened the inhibitory effects of quinidine and azimilide. PMID- 15125691 TI - N-acetylcysteine does not protect HepG2 cells against acetaminophen-induced apoptosis. AB - Acetaminophen in large doses is well-known as hepatotoxic, and early therapy with N-acetylcysteine is frequently life-saving. However, in later stages of acetaminophen poisoning, treatment with N-acetylcysteine is not always effective. Although some of the pathways of acetaminophen toxicity and the effect of N acetylcysteine have been elucidated, in depth information on this process is still lacking. Hepatoma-derived HepG2 cultured cells were exposed to acetaminophen (5 and 10 mM), with or without N-acetylcysteine (5 mM), for 24 and 48 hr. For the assessment of oxidative damage, apoptosis and necrosis, we followed redox status, glutathione content, nuclear fragmentation, phosphatidylserine externalization and ultrastructural changes. Variations in Ca2+ level and number of mitochondrial dense granules were also studied. Acetaminophen treatment of HepG2 cells caused oxidative damage and apoptosis. Significant decrease of cellular redox potential and glutathione content were time- and concentration-dependent. The protective effect of N-acetylcysteine was expressed by an increase of intracellular glutathione and of the level of metabolic reduction of the redox indicator Alamar Blue. The apoptogenic effect of acetaminophen was assessed by flow cytometry of annexin V binding, nuclear hypodiploidity, intracellular Ca2+, as well as by ultrastructural examination. Beyond 24 hr of acetaminophen exposure, necrosis was also noticed. We conclude that acetaminophen-induced oxidative damage in HepG2 cultured cells can be prevented by exposure to N-acetylcysteine. However, apoptosis, either early or late, here demonstrated, is not avoided by exposure to N-acetylcysteine. N Acetylcysteine did not prevent acetaminophen-induced plasma membrane asymmetry, nuclear damage, alterations of Ca2+ homeostasis and ultrastructural changes. PMID- 15125692 TI - Reproductive and cytogenetic toxicity of metronidazole in male mice. AB - The purpose of this study was to examine the effects of metronidazole (500 mg/kg b.wt. daily by gavage for 14 consecutive days) on male fertility, haematopoiesis and genotoxic affinity. Mature male Swiss mice were treated with metronidazole and divided into 3 groups each with 10 animals, examined after 2 weeks, 1 and 2 months from the onset of drug administration. The results demonstrated that metronidazole significantly (P<0.05) decreased the weight of the testes, epididymides and accessory sexual organs (seminal vesicles and prostates) after one month from the onset of treatment. While accessory sexual organ weights were restored after 2 months from onset of treatment, the decrease in testes and epididymides weights persisted until 2 months later. The deleterious effects of metronidazole on reproductive organ weights might be due to a decrease in testosterone level after 2 weeks, and 1 and 2 months from the onset of treatment. Metronidazole induced a significant decrease in motile sperm and an increase in abnormal sperm after 1 month. The viability of sperm was normal after 2 months. Metronidazole induced anaemia characterized by decreased erythrocyte and leukocytic counts, haemoglobin content and haematocrit %. The ability of oral metronidazole administration to induce genotoxic damage in somatic cells of mice was evaluated using mitotic index, micronuclei and chromosomal aberration. A significant reduction in mitotic activity was observed two weeks from the onset of drug administration, restoration occurred after one month. A significant and persistence increase in the frequency of chromosomal aberration and micronucleus was observed at all periods of the experiment. In conclusion, the results of this study indicate that 1) metronidazole (500 mg/kg by gavage) for 14 days caused a harmful effect on male fertility in mice after one and two months from start of administration, 2) metronidazole induced anaemia after one month from start of administration, 3) metronidazole at this high dose level (3 times the therapeutic dose in mice) has the ability to induce genotoxic effects in somatic cells. PMID- 15125693 TI - Administration of procyanidins from grape seeds reduces serum uric acid levels and decreases hepatic xanthine dehydrogenase/oxidase activities in oxonate treated mice. AB - In this study we have investigated the effects of administration of procyanidins from grape seeds on serum uric acid levels in a model of hyperuricaemia in mice pretreated with oxonate, as well as the xanthine dehydrogenase and xanthine oxidase activities in mouse liver in vivo. The procyanidins, when orally administered to the oxonate-pretreated hyperuricaemic mice, were able to elicit a dose-dependent hypouricaemic effect. At a dose of 400 mg/kg for 3 days, the serum urate levels of the oxonate-pretreated mice were not different from the normal mice. In addition, the hepatic activities of xanthine dehydrogenase and xanthine oxidase in the procyanidins-treated mice were found to decrease significantly. However, the hypouricaemic effects observed in the experimental animals did not seem to parallel the changes in xanthine dehydrogenase and xanthine oxidase activities, implying that the procyanidins might be acting via other mechanisms apart from simple inhibition of enzyme activities. Furthermore, the procyanidin treated animals exhibited normal growth while the allopurinol-treated animals exhibited some retarded growth. These results demonstrated for the first time that the procyanidins from grape seeds possess in vivo urate-lowering activities. The potential application of these natural compounds in the treatment of hyperuricaemia is discussed. PMID- 15125694 TI - (+/-) Epibatidine increases acetylcholine release partly through an action on muscarinic receptors. AB - Epibatidine has been used in a wide dose range and was found to produce both nociceptive and antinociceptive effects. The different effects were partly explained by an action on multiple nicotinic receptor systems. The present study investigated the possibility that part of the action of intraspinally or subcutaneously administered (+/-) epibatidine, is mediated through an action on muscarinic receptors. Radioligand receptor assays were performed using homogenates of rat spinal cord and muscarinic M1-M5 receptors expressed in Sf9 cells. The intraspinal acetylcholine releasing effect of intraspinally and subcutaneously administered (+/-) epibatidine was studied with and without with atropine pretreatment. (+/-) Epibatidine has affinity for muscarinic receptors both in spinal cord tissue and expressed in Sf9 cells. The intraspinal administration of 160 microM (+/-) epibatidine produced an increase in acetylcholine release that was reduced by pretreatment with 100 microM atropine. Subcutaneous administration of 30 microg/kg (+/-) epibatidine produced an increase in intraspinal acetylcholine release that was not inhibited by 5 mg/kg subcutaneous atropine pretreatment. We conclude that (+/-) epibatidine, in microM concentrations, is a partial muscarinic receptor agonist that may interact with spinal muscarinic receptors to increase acetylcholine release. Epibatidine induced spinal acetylcholine release observed after subcutaneous administration appears not to be mediated via muscarinic receptor. The dual action on both nicotinic receptors and muscarinic receptors may explain the potent analgesic effect observed after epibatidine administration. PMID- 15125695 TI - Cellular potassium ion deprivation enhances apoptosis induced by cisplatin. AB - The anticancer drug cisplatin induces cell death by apoptosis. Apoptosis is dependent on cellular loss of potassium ions (K+). We have recently shown that the antifungal drug amphotericin B (enhancing K+ efflux), combined with the Na+, K+, 2Cl(-)-cotransport blocker bumetanide (decreasing K+ influx), augmented cisplatin-induced apoptosis in vitro. We therefore quantified K+ fluxes with the K+ analogue rubidium (86Rb+) in cisplatin-induced apoptosis of mesothelioma cells treated with bumetanide and amphotericin B. Bumetanide combined with amphotericin B enhanced cisplatin-induced apoptosis by a pronounced initial reduction of K+ influx due (in addition to Na+, K+, 2Cl(-)-cotransport inhibition) also to Na+, K+, ATPase pump inhibition. As 86Rb+ efflux was initially preserved, combination of the drugs would lead to net K+ loss. Combinations of K+ flux modulators leading to cellular potassium ion deprivation thus augments cisplatin-induced apoptosis and could therefore possibly be used to enhance the antitumour efficacy of cisplatin treatment. PMID- 15125696 TI - Fexofenadine does not affect omeprazole pharmacokinetics: both are putative P glycoprotein substrates. AB - An in vitro study has recently suggested that a proton pump inhibitor, omeprazole, is a modest substrate of P-glycoprotein. Several studies have shown P glycoprotein is involved in the absorption and excretion of fexofenadine. Therefore, we examined the effect of fexofenadine on the pharmacokinetics of omeprazole. Eight healthy volunteers participated in this study. They received a single oral dose of 40 mg omeprazole before and after 60 mg fexofenadine (10 doses over 6 days). Blood samplings were performed up to 8 hr after each dosing. Plasma concentrations of omeprazole and its two metabolites were quantified with high-performance liquid chromatography. In addition, the effect of fexofenadine on P-glycoprotein function was examined by flow cytometry using rhodamine 123 and CD56-positive lymphocytes. Comparison of the pharmacokinetic parameters of omeprazole before and after fexofenadine revealed that there were no differences in peak concentration, time to peak concentration, area under the time concentration curve up to 8 hr, and elimination half-life. There were also no differences in these pharmacokinetic parameters for the two metabolites of omeprazole. Flow cytometric analysis revealed that fexofenadine did not inhibit the efflux of rhodamine 123. This study indicated that there was probably no drug interaction between omeprazole and fexofenadine, which might be due to less contribution of P-glycoprotein to omeprazole absorption, insufficient inhibitory effect of fexofenadine on P-glycoprotein, or the involvement of other transporters such as organic anion transporting polypeptides. PMID- 15125699 TI - Stem Cell Transplantation in Children: Current Results and Controversies. Hilton Head, South Carolina, USA, 6-8 March 2003. Proceedings. PMID- 15125698 TI - Prevention of allergic disease in childhood: clinical and epidemiological aspects of primary and secondary allergy prevention. AB - The development and phenotypic expression of atopic diseases depends on a complex interaction between genetic factors, environmental exposure to allergens,and non specific adjuvant factors, such as tobacco smoke, air pollution and infections. Preventive measures may include both exposure to allergens and adjuvant risk/protective factors and pharmacological treatment. These measures may address the general population, children at risk for development of atopic disease (high risk infants), children with early symptoms of allergic disease or children with chronic disease. The objective for this review was to evaluate possible preventive measures as regards prevention of development of allergic disease in childhood--primary prevention--and also some aspects of the effect of specific allergy treatment as regards secondary prevention in children with allergic asthma and allergic rhinoconjunctivitis. In one prospective observational study of a birth cohort of unselected infants we evaluated possible predictive/risk factors. In two prospective intervention studies including 1 yr birth cohorts of high-risk(HR) infants we investigated the effect of feeding HR infants exclusively breast milk (BM) and/or hydrolyzed cow's milk-based formula the first 4-6 months as regards: (i) the allergy preventive effect of BM/extensively hydrolysed formula (eHF) compared with ordinary cow's milk-based formula, (ii) the effect of two different eHFs, a whey (Profylac) and a casein-based (Nutramigen) formula, as regards development of cow's milk protein allergy (CMA), and (iii) a comparison of the preventive effect of eHF (Profylac/Nutramigen) with a partially hydrolyzed cow's milk-based formula (pHF) (NanHA) as regards development of CMA. None of the mothers had a restricted diet during pregnancy or lactation period. In two prospective randomized intervention studies we evaluated the preventive effect of specific allergen avoidance and specific immunotherapy (SIT) in children with allergic asthma and allergic rhinoconjunctivitis, respectively. The combination of atopic heredity and elevated cord blood IgE resulted in the best predictive discrimination as regards development of allergic disease. The optimal high-risk group was defined by either double parental atopic predisposition or single atopic predisposition, the latter combined with a cord blood IgE > or = 0.3 kU/1. 66% of unselected infants were daily exposed to tobacco smoke, which was a significant risk factor for recurrent wheezing until the age of 1.5 yr. HR infants were breastfed for a longer period and less exposed to tobacco smoke than unselected infants. Exclusively BM/eHF for at least 4 months was associated with a significantly reduced cumulative prevalence of CMA [3.6% (5/141) vs. 20%(15/75) in the control group] up to 5 yr. The effect of the two different eHFs was similar. Exclusively breastfed infants were significantly less exposed to tobacco smoke and pets, had solid foods introduced later and belonged to higher social classes. pHF was significantly (p = 0.05) less effective than eHF as regards prevention of development of CMA. A diet period of 4 months seems to be as efficient as 6 months or more as regards development of CMA. A few ongoing prospective, randomized intervention studies have produced the first indication that avoidance of indoor allergens such as house dust mite (HDM) in HR infants may reduce the incidence of severe wheeze and sensitization during the first 1-4 yr of age. Long-term follow-up is awaited. In a prospective, double blind placebo-controlled study in children with doctors diagnosed asthma and documented HDM allergy, we found that semipermeable polyurethane mattress and pillow encasings (Allergy Control) when compared with placebo encasings resulted in a significant perennial reduction of HDM exposure and a significant reduction in the needed dose of inhaled steroids by approximately 50% (mean dose: 408 microg--227 microg/day) after 1-yr follow-up. In another randomized prospective study we investigated the possible preventive effect of SIT in children with allergic rhinoconjunctivitis and grass/birch pollen allergy as regards development of asthma. Among those without asthma significantly fewer in the SIT group developed asthma when compared with the control group (19/79 = 24% vs.32/72 = 44%) after the first 3 yr; and methacholinebronchial provocation test results improved significant in the SIT group. The results of our studies support the evidence that the risk for development of early allergic manifestations e.g. CMA and atopic dermatitis can be reduced significantly by simple dietary measures for the first4 months of life. In all infants breastfeeding should beencouraged for at least 4-6 months, and exposure to tobacco smoke should be avoided during pregnancy and early childhood. In HR infants a documented hypoallergenic formula (at present eHF) is recommended if exclusive breastfeeding is not possible for the first 4 months. In homes of HR-infants, current evidence supports measures to reduce the levels of indoor allergens. e.g. HDM and pets. In symptomatic children allergen-specific treatment may influence both the symptoms and the prognosis. Allergen avoidance can reduce the need for pharmacological treatment, SIT may have the potential for preventing the development of asthma in children with allergic rhinoconjunctivitis. and it may be possible to interfere with the natural course of allergic diseases. PMID- 15125700 TI - History of pediatric stem cell transplantation. AB - During the past 50 yr, intensive studies into the use of hematopoietic cell transplantation (HCT) for therapy of cancer and non-malignant hematologic diseases have changed this treatment modality from one that was thought to be plagued by insurmountable complications to one that is now standard therapy for some diseases. Continued research by transplant teams worldwide is likely to allow continued progress toward developing novel and improved treatment modalities and even wider application of the use of pluripotent hematopoietic stem cells in the treatment of human diseases. PMID- 15125701 TI - Non-myeloablative allogeneic transplantation -- state-of-the-art. AB - Work by several groups of investigators has brought about changes in the way hematopoietic cell allografts are being done to treat patients with hematologic diseases. Less intensive conditioning regimens have been introduced, and in the case of patients with hematologic malignancies, the burden of eradicating malignant cells has shifted from high-dose chemoradiation treatment toward using the hematopoietic cell donor's T lymphocytes for that purpose by invoking allogeneic graft-versus-tumor effects. While the majority of the transplant regimens used in these efforts are still fairly intense and toxic, a radical departure from conventional transplantation focuses on the almost exclusive use of immunosuppressive agents with little toxicity to establish the allografts. The success of the procedure for patients with hematologic malignancies rests on replacing the host's hematopoietic cells by the allograft. For non-malignant diseases, the procedure can be used to establish a stable state of mixed donor/host hematopoietic chimerism, which, in itself, may be sufficient to cure disease manifestations. PMID- 15125702 TI - Recent advances in the prevention of CMV infection and disease after hematopoietic stem cell transplantation. AB - Cytomegalovirus (CMV) remains an important pathogen in hematopoietic stem cell transplant (HCT) recipients in the current era of antiviral prophylaxis and preemptive therapy, despite the almost complete elimination of CMV disease during the first 3 months after transplantation. Pretransplant CMV serostatus of the donor and/or recipient remains an important risk factor for poor post-transplant outcome, especially in highly immunodeficient patients (e.g. recipients of ex vivo or in vivo T-cell depletion). Prevention of late CMV disease continues to be a challenge in selected high-risk populations, and indirect immunomodulatory effects of CMV (e.g. invasive bacterial and fungal infections) appear to contribute to the poor outcome. The risk of developing antiviral resistance remains low in most patients; however, in a setting of intense immunosuppression (e.g. after transplantation from a haploidentical donor) the incidence may be as high as 8%. Transfusion-transmitted CMV infection can be reduced by the provision of seronegative or leukocyte-depleted blood products; however, a small risk of 1 2% of CMV disease remains. Surveillance and preemptive therapy is effective in preventing transfusion-related CMV disease. The development of new drugs and immunologic strategies (adoptive transfer of CMV-specific T-cells and donor/recipient vaccination strategies) are important goals for the elimination of the negative impact of CMV in the HCT setting. PMID- 15125703 TI - Application of stem cell transplant for brain tumors. AB - Brain tumors are the second most common malignancy in children and the most common solid tumor. The majority of children are treated with surgery alone or in combination with radiation and/or chemotherapy. Recently investigators have used high dose chemotherapy with autologous stem cell rescue (ASCR) in patients with malignant brain tumors. This approach has been most successful in chemosensitive tumors including medulloblastoma, supratentorial primitive neuroectodermal tumors (SPNET) and central nervous system germ cell tumors (CNS GCT). In addition, the use of high dose chemotherapy has enabled the reduction and in many cases elimination of radiation therapy to very young children. To date there have been no prospective randomized studies comparing high dose chemotherapy and ASCR with conventional therapy. Radiation therapy is often not an option for patients with recurrent disease and conventional dose chemotherapy rarely if ever results in long-term survival. Unfortunately, the majority of studies using conventional therapy in order to delay irradiation in young children newly diagnosed with malignant brain tumors have been unsuccessful. Although the numbers are small, preliminary data suggest that not only is survival but also quality of life is superior with the use of high dose chemotherapy. Future studies will most likely include the use of new agents as part of the cytoreduction. In addition, through the use of peripheral blood stem cells and improvements in supportive care, multiple courses of high dose chemotherapy can be administered. High dose chemotherapy with ASCR is a foundation upon which many different types of therapies can be built. Several possibilities include the use of anti angiogenesis agents, monoclonal antibodies and biologic response modifiers. PMID- 15125704 TI - Sickle cell anemia and hematopoietic cell transplantation: When is a pound of cure worth more than an ounce of prevention? AB - Hematopoietic cell transplantation (HCT) is curative therapy for sickle cell anemia (SCA). However, its widespread use is constrained by donor availability and by concerns about its short-term and long-term toxicities. Current efforts to identify suitable candidates for HCT to decrease the toxicity of HCT, and to broaden its availability are discussed. PMID- 15125705 TI - Endocrine complications of high-dose therapy with stem cell transplantation. AB - Evaluations of endocrine function following hematopoietic cell transplantation demonstrate that the endocrine function abnormalities observed are related to the type of transplant preparative regimen received. Children given high dose cyclophosphamide (CY) only have normal thyroid function, normal growth and development. Children who received a busulfan (BU) plus CY preparative regimen usually have normal thyroid function, normal prepubertal growth, delayed or absent pubertal development, and blunted post-pubertal growth. Recipients of preparative regimens containing total body irradiation may be anticipated to have some thyroid dysfunction, impaired growth rates and delayed or absent pubertal development. Post-pubertal teens and young adults are likely to have gonadal function recover if they received a preparative regimen with CY only but are likely to have primary gonadal failure if they received a preparative regimen with BU or total body irradiation. Individuals whose gonadal function becomes normal have become parents of normal children. All patients who receive a marrow transplant should be followed long-term for development of endocrine function abnormalities. PMID- 15125706 TI - Strategic approaches to osteoporosis in transplantation. AB - Recipients of stem cell transplantation are at risk for osteopenia and osteoporosis. Longitudinal studies performed in adults have shown that significant bone demineralization occurs following myeloablative therapy and subsequent immune suppression. Among children and adolescents, cross-sectional analyses indicate that younger patients are also at risk for long-term bone toxicity. Strategies to detect and manage this disorder in pediatric SCT recipients are presented. PMID- 15125707 TI - Detection and treatment of minimal residual disease in high-risk neuroblastoma. AB - Intensive, myeloablative therapy supported by autologous hematopoietic stem-cell transplantation (AHSCT) has improved the outcome for children with high-risk neuroblastoma. However, >50% of patients develop recurrent neuroblastoma, often from minimal residual disease (MRD). Immunocytological and reverse transcriptase polymerase chain reaction (RT-PCR) for genes highly expressed in neuroblastoma both can detect small amounts of MRD in blood and bone marrow, and detection of MRD at certain levels during therapy has prognostic value. Radionucleotide scans using meta-iodobenzaguanidine (MIBG) imaging allows sensitive detection of neuroblastoma in patients, but whether or not all MIBG-positive disease detected after AHSCT will progress remains to be defined and is complicated by use of post AHSCT therapy. Selective removal of tumor cells from marrow or blood stem cells harvested for AHSCT could decrease recurrence by preventing infusion of tumorigenic cells with AHSCT. Treating MRD after AHSCT with the differentiation inducing retinoid 13-cis-retinoic acid significantly /improved EFS of high-risk neuroblastoma patients. Randomized clinical trials in the Children's Oncology Group are testing the value of purging blood stem cells and also whether post AHSCT therapy with an anti-GD2 monoclonal antibody (combined with cytokines) improves outcome over use of 13-cis-retinoic acid alone. New approaches to treating neuroblastoma MRD that are in early clinical trials include the cytotoxic retinoid fenretinide and the hu14.18-IL2 immunocytokine. It is anticipated that testing novel approaches to treating neuroblastoma MRD will be the subject of future phase-III randomized trials. PMID- 15125708 TI - Treatment of advanced Ewing tumors by combined radiochemotherapy and engineered cellular transplants. AB - This review will focus primarily on own recent work on the treatment of advanced Ewing tumors (AETs) and will attempt, in addition, to give a comprehensive overview of novel developments. The field under review has been shaped by investigators from both Europe and the United States of America in a scientific debate evolving over more than a decade at the meetings of the International Society of Pediatric Oncology and other scientific meetings. In the light of this debate, most oncologists will agree that patients with AETs are facing the worst prognosis of all patients with this disease and include both: (i) patients with primary metastatic disease with the worst prognosis as well as (ii) patients with relapse with the worst prognosis. The contributions of various investigators have lead to the identification of specific risk stratification criteria to overcome the heterogeneity of patients within the conventionally defined clinical stages of localized metastatic and relapsed disease. This review will address the following issues of treatment of AETs: (i) a definition of AET; (ii) risks and benefits of allogeneic vs. autologous stem cell transplantation; (iii) the role of total body irradiation; (iv) the number of involved bones as a risk factor in multifocal bone disease in AET; (v) the development of immunogene therapy in AET; (vi) the matching of radiochemo- and immunotherapy in AET; (vii) the future perspective of functional genomics and targeted therapy. PMID- 15125709 TI - Megatherapy and stem cell transplantation for Ewing's family of tumors: a critical review of current literature. AB - Although a multimodal approach consisting of chemotherapy and local control with surgery and/or radiation has improved survival in children with Ewing's sarcoma family of tumors, the prognosis for patients with high-risk disease remains poor. More aggressive treatments with high-dose myeloablative chemotherapy followed by stem cell rescue have been utilized in an attempt to improve survival in these patients. Although many studies have been published, it is difficult to interpret the data since patient populations were heterogeneous with respect to disease stage, prior therapy, conditioning and stem cell source. Furthermore, there was no uniform definition of high risk, the sample sizes were small and most studies lacked appropriate control groups. Assessment of the utility of megatherapy will require prospective controlled studies. PMID- 15125710 TI - Alternate site biventricular pacing: Bi-V in the RV--is there a role? PMID- 15125711 TI - Noncontact mapping of the left ventricle: insights from validation with transmural contact mapping. AB - It is not clear whether the noncontact electrograms obtained using the EnSite system in the left ventricle resemble most closely endocardial, intramural, or epicardial contact electrograms or a summation of transmural electrograms. This study compared unipolar virtual electrograms from the EnSite system with unipolar contact electrograms from transmural plunge needle electrodes using a 256-channel mapping system. The study also evaluated the effects of differing activation sites (endocardial, intramural, or epicardial). A grid of 50-60 plunge needles was positioned in the left ventricles of eight male sheep. Each needle had four electrodes to record from the endocardium, two intramural sites, and the epicardium. Correlations between contact and noncontact electrograms were calculated on 32,242 electrograms. Noncontact electrograms correlated equally well in morphology and accuracy of timing with endocardial (0.88 +/- 0.15), intramural (0.87 +/- 0.15), epicardial (0.88 +/- 0.15), and transmural summation contact electrograms (0.89 +/- 0.14) during sinus rhythm, endocardial pacing, and epicardial pacing. There was a nonlinear relationship between noncontact electrogram accuracy as measured by correlation with the contact electrogram and distance from the multielectrode array (MEA): beyond 40 mm accuracy decreased rapidly. The accuracy of noncontact electrograms also decreased with increasing distance from the equator of the MEA. Virtual electrograms from noncontact mapping of normal left ventricles probably represent a summation of transmural activation. Noncontact mapping has similar accuracy with either endocardial or epicardial sites of origin of electrical activity provided the MEA is within 40 mm of the recording site. PMID- 15125712 TI - Postfibrillatory enhancement of left atrial contractility after short paroxysms of atrial fibrillation. AB - Implantable cardioverter defibrillators and pacemakers detect an increasing number of silent episodes of AF. In a porcine model, the study evaluated the contractility of the left atrial appendage (LAA) during AF paroxysms as they may occur in patients. Peak outflow velocity of the LA and mean outflow velocity of the LAA (LAA-V(outmean)) (n = 17) were measured before, during, and after induction of self-terminating AF. LAA-V(outmean) was also measured during incremental pacing from different atrial sites using epicardial Doppler probes (n = 6) and during continuous recordings (n = 5) of 40 minutes of pacing maintained AF. Compared to baseline sinus rhythm, LAA-V(outmean) increased during short AF episodes (41 +/- 3 vs 35 +/- 2 cm/s, P < 0.05). After termination of the AF episodes, LAA-V(outmean) further increased (69 +/- 15 cm/s, P < 0.001 vs baseline). This "postfibrillatory enhancement" maintained after repeated induction of short AF paroxysms. During prolonged AF episodes lasting 40 minutes, an initial hypercontractility (44 +/- 2 vs 38 +/- cm/s, P < 0.01) was followed by a hypocontractility after 20 minutes (29 +/- 12 P < 0.05 vs SR) and a postfibrillatory enhancement after cessation of AF (56 +/- 12 vs 27 +/- 9 cm/s at 40 minutes AF, P < 0.001). L-type Ca channel blockade abolished the initial hypercontractility during AF and the postfibrillatory enhancement. Repetitive AF paroxysms up to 2 minutes did not decrease left atrial contractility. During maintained AF up to 40 minutes an initial hypercontractility and a consecutive hypocontractility, which is overcompensated by a postfibrillatory enhancement of atrial inotropy after cessation of AF, are present. The observed phenomenon seems to be related to an increased Ca(2+) influx through the L-type Ca(2+) channel. PMID- 15125713 TI - A comparison of biventricular and conventional transvenous defibrillation: a computational study using patient derived models. AB - Conventional transvenous defibrillation is performed with an ICD using a dual current pathway. The defibrillation energy is delivered from the RV electrode to the superior vena cava (SVC) electrode and the metallic case (CAN) of the ICD. Biventricular defibrillation uses an additional electrode placed in the LV free wall with sequential shocks to create an additional current vector. Clinical studies of biventricular defibrillation have reported a 45% reduction in mean defibrillation threshold (DFT) energy. The aim of the study was to use computational methods to examine the biventricular defibrillation fields together with their corresponding DFTs in a variety of patient derived models and to compare them to simulations of conventional defibrillation. A library of thoracic models derived from nine patients was used to solve for electric field distributions. The defibrillation waveform consisted of a LV --> SVC + CAN monophasic shock followed by a biphasic shock delivered via the RV --> SVC + CAN electrodes. When the initial voltage of the two shocks is the same, the simulations show that the biventricular configuration reduces the mean DFT by 46% (3.5 +/- 1.3 vs 5.5 +/- 2.7 J, P = 0.005). When the leading edge of the biphasic shock is equal to the trailing edge of the monophasic shock, there is no statistically significant difference in the mean DFT (4.9 +/- 1.9 vs 5.5 +/- 2.7 J, P > 0.05) with the DFT decreasing in some patients and increasing in others. These results suggest that patient-specific computational models may be able to identify those patients who would most benefit from a biventricular configuration. PMID- 15125714 TI - Deep myocardial ablation lesions can be created with a retractable needle-tipped catheter. AB - RF catheter ablation of ventricular tachycardia is sometimes limited by inadequate lesion depth. This study investigated the use of a retractable needle tipped catheter to create deep RF lesions in vivo in porcine myocardium. An 8 Fr electrode catheter with an extendable 27-gauge needle at the tip was modified for RF ablation by embedding a thermocouple and attaching a pin connector. In three swine (32-58 kg) the left ventricle was entered via the femoral artery and endocardial contact was made. The needle was advanced 10 mm and 13 RF applications were made under a controlled temperature (90 degrees C x 120 s). Nine control lesions were made using a standard 4-mm tip catheter (60 degrees C x 120 s). The lesions were fixed, serially sectioned from the endocardium, digitally imaged, and quantified. Needle ablation lesions were deeper (10.15 +/- 0.77 vs 5.67 +/- 0.37 mm, P < 0.001) and more likely to be transmural (77 vs 11%, P = 0.008) than control lesions. The volume of control lesions, however, was larger (358.4 +/- 56.2 vs 174.7 +/- 18.6 mm(3), P = 0.002) due to a significantly larger cross-sectional area at the endocardium (0.548 +/- 0.04 vs 0.151 +/- 0.01 cm(2), P < 0.001). At depths > 6 mm, the needle electrode lesions had a greater cross-sectional area (0.136 +/- 0.01 vs 0.005 +/- 0.004 cm(2), P < 0.001). Catheter-based needle ablation is feasible and allows creation of deeper lesions that can be transmural. Although deep, the lesions had a small cross-sectional area such that precise targeting would be required for success. PMID- 15125715 TI - Pacemaker syndrome in the young: do children need dual chamber as the initial pacing mode? AB - The aim of this study was to determine if single chamber rate responsive ventricular pacing (VVIR) predisposes growing children to develop pacemaker syndrome (PS), and if so, what are determining factors and/or clinically useful predictors. PS is a constellation of symptoms that result from the lack of consistent AV sequential filling due to atrial contraction against closed AV valves. PS has not been commonly reported in the young. Data from all patients with pacemakers with congenital complete atrioventricular block (CAVB) with normal anatomy, and those with congenital heart disease (CHD), and surgically acquired CAVB were reviewed. Inclusion criteria were normal ventricular function by cardiac ultrasound and 100% VVIR pacing. Of 89 patients with VVIR pacemaker implants, 33 met these criteria. Of these, 19 developed PS. For statistical analysis, chi-square and independent samples t-test was used with significance defined at P < or = 0.05. No consistent association was found between cardiac anatomy, type of CAVB, or age at implant with development of PS. However, PS did correlate with duration of pacing (P = 0.02). The exercise stress test showed significant differences between 100% VVIR-paced patients with and without PS, in terms of work rate (P = 0.002) and measured oxygen consumption (P = 0.01). This study shows that PS appears to be a time related event in younger children with normal ventricular function who are 100% ventricular paced. Thus, this supports VVIR pacing as an adequate and cost-effective initial therapy for symptomatic bradycardia due to CAVB. PMID- 15125716 TI - Sinus and paced P wave duration and dispersion as predictors of atrial fibrillation after pacemaker implantation in patients with isolated sick sinus syndrome. AB - The aim of this study was to prospectively evaluate the sinus and the paced P wave duration and dispersion as predictors of AF after pacemaker implantation in patients with isolated sick sinus syndrome (SSS). The study included 109 (69 women, mean age 72 +/- 11 years) patients with SSS, 59 with bradycardia tachycardia syndrome (BTS). A 12-lead ECG was recorded before pacemaker implantation and during high right atrial and septal right atrial pacing at 70 and 100 beats/min. The ECGs were scanned into a computer and analyzed on screen. The patients were treated with AAIR (n = 52) or DDDR pacing. The P wave duration was measured in each lead and mean P wave duration and P wave dispersion were calculated for each ECG. AF during follow-up was defined as: AF in an ECG at or between follow-up visits; an atrial high rate episode with a rate of > or =220 beats/min for > or =5 minutes, atrial sensing with a rate of > or =170 beats/min in > or =5% of total counted beats, mode-switching in >/=5% of total time recorded, or a mode switching episode of > or =5 minutes recorded by the pacemaker telemetry. The ECG parameters were correlated to AF during follow-up. Mean follow-up was 1.5 +/- 0.9 years. None of the ECG parameters differed between patients with AF and patients without AF during follow-up, nor was there any difference between groups after correction for BTS and age. BTS was the strongest predictor of AF during follow-up (P < 0.001). P wave duration and dispersion measured before and during pacemaker implantation were not predictive of AF after pacemaker implantation in patients with isolated SSS. PMID- 15125717 TI - Is isolated congenital heart block associated to neonatal lupus requiring pacemaker a distinct cardiac syndrome? AB - Isolated congenital heart block (ICHB) is frequently associated with neonatal lupus syndrome (NLS). Therefore few data are available regarding the long-term cardiac outcome of newborns with ICHB and the pathogenic mechanisms are not yet defined. In order to compare demographic features and cardiological outcome of patients with ICHB submitted to pacemaker (PM) implantation with and without NLS, forty ICHB patients were evaluated pre- and post-PM implantation, by clinical, electrocardiogram, Holter Monitoring, treadmill test, and electrophysiological study. According to the presence of antibodies to 52 and 60 kDa Ro/SSA and La/SSB proteins in mother's sera, it was found that 60% (24/40) of patients had ICHB associated to NLS (ICHB/NL+). Twenty-three of 24 ICHB/NL+ patients were asymptomatic, and 16 (67%) were female (P = 0.013). The frequency of syncope, mitral insufficiency (MI), and congestive heart failure (CHF) was similar pre-PM implantation in both ICHB/NL+ and ICHB/NL- groups (P > 0.05). After PM implantation, MI and CHF were only observed in ICHB/NL+ patients, although not statistically significant. Interestingly, 67% of ICHB/NL+ were noticed before one year of age while only one fourth of ICHB/NL- was diagnosed in this period (P = 0.024). Almost half (46%) of ICHB/NL+ patients required PMs in the first 24 months of life, whereas only one in the ICHB/NL- received a PM at the same age (P = 0.02). In ICHB patients requiring PM implantation, the antibody-mediated lesion seems to be associated with an earlier onset and a more severe heart disease, in spite of the uniform criteria for PM indication. PMID- 15125718 TI - Detection of acute myocardial ischemia during percutaneous transluminal coronary angioplasty by endocardial acceleration. AB - The first heart sound is generated by vibrations from the myocardium during isovolumic contraction. Peak endocardial acceleration (PEA) has been used previously to measure these vibrations in humans and correlates with myocardial contractility during inotropic interventions. It is unknown if changes in PEA can be used to characterize a reduction in contractility during ischemic episodes. This study was designed to evaluate the use of an endocardial accelerometer for the detection of acute myocardial ischemia. Thirteen patients undergoing routine percutaneous transluminal coronary angioplasty (PTCA) consented to having a single-axis, lead-based accelerometer positioned in the right ventricular apex. PEA was defined as the maximum peak-to-peak amplitude during a window 50 ms before to 200 ms following the peak R wave. Time of endocardial acceleration (TEA) was defined as the time from the peak R wave to the maximum accelerometer signal within this window. To obtain a more robust estimate of the strength of vibrations, a 100-beat template of the accelerometer signal was constructed at baseline and applied as a matched filter during ischemia. The peak magnitude of the filtered endocardial accelerometer signal (Max Filtered EA) was used as an index of signal intensity. Median baseline PEA, TEA, and Max Filtered EA were 0.91 +/- 0.35 g, 75.2 +/- 16.2 ms, and 0.40 +/- 0.20 g, respectively. PEA and Max Filtered EA significantly decreased by 7% during ischemia (0.91 to 0.85 g and 0.40 to 0.37 g, both P < 0.05, respectively). TEA did not significantly change from baseline (77.0 ms, P = ns). The results of this study suggest that acute ischemia can be detected with an endocardial accelerometer in humans. PMID- 15125719 TI - Performance of left ventricular versus biventricular pacing in chronic heart failure assessed by stress echocardiography. AB - Acute hemodynamic studies suggest that resynchronization therapy using single site left ventricular pacing (LVP) is equivalent to biventricular pacing (BIVP). The aim of this study was to assess the performance of LVP versus BIVP during exercise by means of stress echocardiography. A total of 28 patients (25 men and 3 women, mean age 60.9 +/- 8 years) with advanced chronic heart failure and impaired ventricular conduction (QRS > 150 ms) were studied. Patients were randomly allocated to either BIVP or LVP mode with a crossover on the next day and cardiac output was estimated at rest and during each stage of bicycle ergometry in supine position by means of velocity time integral formula. Maximum exercise level was comparable for both pacing modes (up to 100 W) and no significant differences were revealed either in heart rate or in blood pressure at rest and during any step of exercise. LVP was associated with significantly higher cardiac output at rest (3.2 +/- 0.5 vs 2.8 +/- 0.6 l/min, P < 0.01) and during low level exercise (4.4 +/- 0.8 vs 3.9 +/- 0.8 l/min at 25 W, P < 0.05) as compared with BIVP. There was a trend towards higher cardiac output for LVP even at higher levels of exercise. These effects were predominantly confined to patients with idiopathic dilated cardiomyopathy. It is concluded that cardiac resynchronization therapy using single-site LVP results in better hemodynamic response as compared with BIVP, both at rest and during physical exercise. PMID- 15125720 TI - Atrioventricular block during upright tilt table test. AB - Patients with a cardioinhibitory response (asystole or atrioventricular block [AVB]) during upright tilt table test (UTT) constitute a therapeutic challenge. Our present knowledge is partial and in those who experience AVB is absent. Furthermore, we ignore if there is any difference between both groups, particularly pacemaker indication. We aimed to study patients with a cardioinhibitory response during UTT and incidence of AVB during UTT, compared to asystole; plus the outcome during prolonged follow-up. Of 867 patients who underwent UTT, 172 were positive for a neurally mediated response, all with normal neurocardiovascular evaluation. Of the 172 patients, 6 (3.4%) developed AV block (group A), and 26 (15.1%) experienced asystole (group B). Group A included 6 women (100%), mean age 21 +/- 12 years. All patients in group A had sinus rate deceleration during AVB. Group B included 10 women (38%), mean age 28 +/- 17 years, and a mean pause of 15 +/- 9 seconds. We contacted 30 of 32 patients, mean follow-up of 45 +/- 38 months. Seven patients in group B had syncopal recurrences; five had 2 or more episodes. One patient from group B received a DDD pacemaker. In group A, one had one recurrence. No deaths were observed. AVB during UTT is rare, occurs in young women, and is always associated with sinus rate deceleration. Medium- to long-term prognosis is good, and equivalent to patients with asystole. There is no evidence that patients with AVB during UTT require a pacemaker implant. PMID- 15125721 TI - The use of a modified sphygmomanometer to detect atrial fibrillation in outpatients. AB - This study was designed to assess the accuracy of a modified sphygmomanometer, that measures pulse irregularity, to detect atrial fibrillation (AF). An irregularity index, defined as the standard deviation of the time intervals between beats divided by the mean of the time intervals, was used to analyze standard 12-lead ECGs from hospitalized patients. A threshold irregularity index was selected such that all ECGs with AF exceeded this irregularity index value. A modified automatic blood pressure monitor was designed to detect AF by calculating the irregularity index of the pulse. The device was used to calculate the irregularity index in an unselected group of outpatients during scheduled office visits in which a standard 12-lead ECG was performed. A total of 125 ECGs, 53 with AF, were analyzed. Using a threshold irregularity index of 0.066, the sensitivity for detecting AF was 100%, the specificity was 92% and diagnostic accuracy 95%. A modified sphygmomanometer was used to analyze the pulse of 450 outpatients, 54 of whom were documented by ECG to be in AF. When paired readings were analyzed, the rhythm was considered to be irregular if both readings were greater than the threshold index. Using a threshold index of 0.06, all the AF patients were correctly identified while 37 non-AF patients also exceeded the threshold irregularity index. In this analysis, the sensitivity was 100%, the specificity 91%, and the diagnostic accuracy 92% for detecting AF. The irregularity index determined using a modified sphygmomanometer can accurately identify AF. PMID- 15125722 TI - Abnormal right atrial electrograms predict the transition to chronic atrial fibrillation in paced patients with sick sinus syndrome. AB - Although pacing therapy for sick sinus syndrome (SSS) is established, the risk of developing chronic atrial fibrillation (CAF) makes pacing therapy infeasible in some patients. We evaluated whether electrophysiological characteristics of atrial muscle can serve as predictors of the transition to CAF after pacemaker implantation in patients with SSS. Eighty-nine patients with SSS underwent electrophysiological study before pacing therapy. Catheter mapping of 12 right atrial sites was performed during sinus rhythm during electrophysiological. An abnormal atrial electrogram was defined as having a duration of 100 ms or longer, or eight or more fragmented deflections, or both. Right atrial extrastimulation was also performed for atrial vulnerability. After electrophysiological study, all patients underwent pacemaker implantation and were followed up. During the follow-up period of 85 +/- 50 months, development of CAF was observed in 12 patients (group A). The remaining 77 patients remained in sinus rhythm (group B). There were significantly more abnormal atrial electrograms in group A than group B (2.7 +/- 2.3 vs 0.8 +/- 1.2; P < 0.001). The distribution of abnormal atrial electrograms was also greater in group A; patients in group A had more abnormal atrial electrograms than patients in group B in both the high and middle right atrium (P < 0.005 and P < 0.01, respectively). Kaplan-Meier analysis showed that almost 50% of the paced patients with abnormal atrial electrograms (n = 42) developed CAF (P < 0.005). Our data suggest that the existence of abnormal atrial electrograms is predictive of the transition to CAF in paced patients with SSS. PMID- 15125723 TI - Basic assessment of paced activation sequence mapping: implications for practical use. AB - Some experiences support the use of atrial paced activation sequence mapping, but there is no systematic study assessing its spatial resolution, reproducibility, and influence of pacing parameters. The aim of this study was to evaluate these issues by using a 24-pole catheter positioned at the atrial aspect of the tricuspid and mitral annuli in 15 patients. Bipolar pacing was performed at two sites (right and left atria), 2 cycle lengths (300 and 500 ms) and two outputs (twice and tenfold the late diastolic threshold voltage for 2-ms pulses). The elapsed time between the atrial activation at the two dipoles adjacent to the pacing dipole (activation time [AT]) was measured during each pacing sequence. Changes in cycle length did not modify the AT. The increase in voltage slightly modified the AT (maximum -2 ms at the RA; 95% CI -3 to -1 ms) due to a greater shortening of the conduction time to the dipole located next to the anode. The 95% limits of the intraobserver and interobserver agreements in the AT measurement were -2 to 3 ms and -3 to 3 ms, respectively. The spatial resolution was studied in ten patients by measuring the AT during pacing from each dipole of a 20-pole catheter with a 1-3-1 mm interelectrode distance. The mean AT change was 10 +/- 4 ms per 6 mm of pacing site displacement (95% CI 8-11 ms, range 2.5 20 ms). In conclusion, paced atrial activation sequence analysis is reproducible, accurate, and relatively independent of pacing parameters. PMID- 15125724 TI - Insertable loop recorder use for detection of intermittent arrhythmias. AB - The advent of prolonged monitoring with the implanted loop recorders has revolutionized the quest for detection of elusive infrequent arrhythmias in patients with unexplained syncope. The capability of prolonged monitoring has permitted us to obtain symptom rhythm correlation in the majority of patients suspected to have underlying infrequent arrhythmia. The implanted loop recorder is easily implanted in the left pectoral region with a minimally invasive procedure, providing at least 14 months of continuous monitoring that is both patient and automatically activated. Several recent studies suggest that it plays a major role in patients with infrequent symptoms and suspected arrhythmia, including patients with syncope and conduction disturbances, mild to moderate underlying heart disease, and atypical epilepsy. In a randomized trial, the device was found to be cost-effective and improved diagnostic yield compared to conventional tilt and electrophysiological testing. Wider application of prolonged monitoring is ongoing, including assessment of ventricular arrhythmias, atrial fibrillation, and conduction disturbances. The implantable loop recorder is most useful in patients with infrequent unexplained syncope when noninvasive testing is negative. PMID- 15125725 TI - Atrial flutter: typical or atypical? PMID- 15125726 TI - Honor thy dog. PMID- 15125727 TI - Successful radiofrequency catheter ablation of an anteroseptal (superoparaseptal) atrioventricular accessory pathway from the left ventricular outflow tract. AB - This case report describes a patient with Wolff-Parkinson-White syndrome in whom the ECG exhibited a typical pattern of an anteroseptal (superoparaseptal) accessory pathway. Successful radiofrequency catheter ablation was achieved from the septal side of the left ventricular outflow tract. It might be worthwhile to map the left side of the anterior septum if an accessory pathway potential is not appreciable along the tricuspid annulus to avoid the potential complication of AV block in patients with a typical anteroseptal accessory pathway ECG pattern. PMID- 15125728 TI - Pacing in high field cardiac magnetic resonance imaging:. AB - Currently, cardiac MRI is contraindicated in patients with an implanted pacemaker or ICD due to safety hazards. However, MRI is promising to play a key role in cardiac diagnostics in near future. This study examined a rat with an implanted pacemaker pacing at a rate of 460/min with high field cardiac MRI. This study showed that pacing during cardiac imaging at 7 Tesla was possible. The pacemaker program was not disturbed by the high field or the strong gradients (maximum dB/dt 400 mT/s). The only noticeable effect on the MRI signal was a signal void of 2 cm around the device. PMID- 15125729 TI - Ventricular fibrillation without overt cardiomyopathy as first presentation of organic cation transporter 2-deficiency in adolescence. AB - This case report describes ventricular fibrillation without overt cardiomyopathy as the presenting symptom of primary carnitine deficiency due to organic cation transporter 2 (OCTN2)-deficiency in a 15-year-old girl. Normally this disease presents early in life with hypoketotic hypoglycemia, muscle weakness, and/or cardiomyopathy. The patient fully recovered after carnitine supplementation. Recognition of this disease is important because its treatment is easy and effective. PMID- 15125730 TI - Inappropriate therapy from a defibrillator complicating transcoronary ablation of septal hypertrophy in a patient with hypertrophic obstructive cardiomyopathy. AB - This case report describes a patient with obstructive hypertrophic cardiomyopathy who received therapy inappropriately from his implanted defibrillator, subsequent to transcoronary alcohol ablation for septal hypertrophy (TASH). Widening of the intracardiac electrogram postablation resulted in "double counting" of the intrinsic ventricular electrogram by the device and inappropriate tachycardia detection. PMID- 15125731 TI - Hyperkalemia induced T wave oversensing leading to loss of biventricular pacing and inappropriate ICD shocks. AB - Inappropriate ICD shocks remain a common problem. Double counting of single ventricular events can occur with biventricular ICDs implanted before univentricular sensing was available. Often this is due to a tachyarrhythmia or loss of left ventricular capture. This report describes a patient who developed hyperkalemia during hemodialysis, received inappropriate ICD shocks and experienced loss of biventricular pacing due to T wave rather than QRS double counting. Oversensing was abolished by reducing the potassium content of the dialysis bath. This underscores the need for careful interpretation of saved electrograms to determine the cause for, and appropriate treatment of, device related problems. PMID- 15125732 TI - Massive transient pulmonary air embolism during pacemaker implantation under mild sedation: an unrecognized hazard of snoring. AB - We present a case of a massive pulmonary air embolism during permanent pacemaker lead implantation under mild sedation in a 73-year-old woman. We used a peel-away sheath that is devoid of a hemostatic valve. The air embolism occurred after deep inspiration associated with loud snoring sounds and resolved after short chest massage and administration of adrenaline. Temporary closure of the intravenous entrance route of the peel-away sheath before inserting the lead may prevent this potentially lethal complication. PMID- 15125733 TI - Implantation of an epicardial dual chamber ICD following unsuccessful percutaneous extraction of a failed ventricular shocking electrode. AB - This report describes the case of a 22-year-old patient presenting with ICD electrode failure where attempted percutaneous electrode extraction was unsuccessful. In view of the patient's young age, an operative approach was considered most appropriate. The failed electrode was surgically extracted and a new epicardial pacing/defibrillator system inserted. PMID- 15125734 TI - Inappropriate sensing of pectoral myopotentials by an implantable cardioverter defibrillator system. AB - A case of inappropriate detection of pectoral myopotentials by an implantable cardioverter defibrillator (ICD) system due to inadvertent reversal of the high voltage pins in the ICD header is reported. Inappropriate high voltage connections can result in malfunction of both sensing and defibrillation functions of some ICDs. PMID- 15125735 TI - Shock coil failure secondary to external irradiation in a patient with implantable cardioverter defibrillator. AB - The use of implantable cardioverter defibrillator (ICD) in the management of malignant ventricular arrhythmia is well established. Radiation treatment is common in malignant neoplasms, but the direct effect of irradiation in ICD is largely not well understood. We describe a case where radiation treatment probably led to shock coil failure. PMID- 15125736 TI - Unsuccessful rescrewing of a dislodged screw-in atrial lead. AB - We report a case of unsuccessful rescrewing of an atrial lead after its early dislodgment in a 64 years old patient because of entrapped endocardial tissue in the screw-in system. PMID- 15125737 TI - Cardiac pacing: how it started, where we are, where we are going. PMID- 15125739 TI - Antibiotics and the risk for breast cancer: a new dilemma. PMID- 15125740 TI - Occult metastases in axillary lymph nodes as a predictor of survival in node negative breast carcinoma with long-term follow-up. AB - Increased detection rate in the lymph nodes is seen with serial sectioning or immunohistochemistry (IHC), but the importance of occult metastases is not resolved. IHC is still not recommended in routine examination of lymph nodes. Axillary lymph nodes from 385 node-negative breast cancer patients with a median follow-up of 25 years were examined with IHC for cytokeratins, applied on routine sections. The association between classic histopathologic prognostic factors and the presence of occult metastases was evaluated. Metastases were found in 45 of 385 cases (12%), 21 metastases (47%) measured < or =0.2 mm, 8 (18%) were larger than 2 mm; 14 metastases were located in the subcapsular sinus, 22 in the parenchyma of the lymph node; and 51% (23/45) of the metastases were recognized on hematoxylin-eosin staining on "second look." The detection of metastases was significantly associated with the number of sectioned lymph nodes (6% metastases for one to five lymph nodes examined versus 17% for more than five lymph nodes) and with histologic subtype (metastases in 11% of the ductal versus 33% of the lobular carcinomas). No significant association was found between occult metastases and age, tumor size, histologic grade, estrogen or progesterone receptor status, p53, or c-erbB-2. Metastases larger than 2 mm predicted a poorer recurrence-free survival rate for the whole series. A subcapsular location of the metastases was a strong predictor of overall survival. Whether or not the metastases could be identified on hematoxylin-eosin sections did not have any prognostic significance. In the multivariate analysis, histologic grade, tumor size of the primary tumor, progesterone receptor status, and the presence of occult metastasis in the lymph nodes had a prognostic impact on survival with a 25-year follow-up. PMID- 15125741 TI - Bone marrow micrometastases and adjuvant treatment of breast cancer. AB - The immunohistochemical detection of epithelially derived cells in the bone marrow of patients with primary breast cancer has been shown to be associated with increased risk of distant relapse as well as higher rates of cancer-related death. Despite the correlation between bone marrow micrometastases and poor outcome in breast cancer patients, bone marrow status does not yet have an established role in patient management. In this prospective study, adjuvant therapy recommendations for 43 patients with stage I, II, or III breast cancer treated with lumpectomy or mastectomy, sentinel lymph node biopsy and/or axillary dissection, and intraoperative bone marrow aspiration were recorded. Recommendations were made by a multidisciplinary tumor board both blinded and unblinded to the results of the bone marrow aspiration. In our study, 10 of the 43 breast cancer patients were found to have bone marrow micrometastases. Four of these patients (40%) had axillary lymph node metastases. When blinded to the results of the bone marrow aspiration, the tumor board recommended adjuvant chemotherapy for these four node-positive patients, as well as two node-negative patients. When unblinded to the results of the bone marrow aspiration, the tumor board did not change its recommendations for any of these six patients. The remaining four node-negative, bone marrow-positive patients were not advised to have adjuvant chemotherapy by the tumor board when blinded to bone marrow status. However, once the tumor board was informed of the presence of bone marrow micrometastases, adjuvant chemotherapy was recommended for all of these patients. The results of this pilot study indicate that the presence of bone marrow micrometastases in breast cancer patients with stage I, II, or III disease does influence recommendations for adjuvant chemotherapy, particularly in patients with node-negative disease. PMID- 15125742 TI - The optimal number of sentinel lymph nodes for focused pathologic examination. AB - The sentinel lymph node (SLN) procedure provides an alternative method for assessing the axillary lymph nodes in patients with breast cancer. The SLN is typically subjected to a focused pathologic examination involving the examination of multiple tissue levels and/or keratin immunohistology. The number of SLNs submitted may vary widely, in some cases rivaling that of a complete axillary dissection (CAD). We examined our experience over the last 2 years in order to determine the optimal number of SLNs for focused pathologic evaluation. All SLN cases for the years 2000 and 2001 were retrieved from the files of the Pathology Department at Magee-Womens Hospital and were tabulated to determine the average number of SLNs per case, the number of SLNs submitted, the actual SLN that was positive for each case, the type of metastasis, and the average number of SLNs per case for each surgeon. There were 662 operative cases that yielded 1576 SLN accessions and 1758 total SLNs. The range of SLNs submitted was 1 to 11. Overall there was a mean of 2.4 SLNs accessioned per case and a mean of 2.7 SLNs per case. A study of the statistics of SLNs submitted by seven surgeons yielded two distinct groups, with one group submitting virtually all of the cases where there were consistently more than four SLNs per case. Ninety-seven percent of positive SLNs were discovered in the first three SLNs submitted, regardless of surgeon identity. The SLNs beyond numbers one to three yielded positive results by keratin in only four cases. Focused pathologic examination of SLNs was most effective for the first three SLNs submitted for any given case. The variation in the number of SLNs submitted per case was different based upon the different practice patterns of surgeons. It is suggested that for more than three SLNs submitted, simple routine lymph node examination would be appropriate. PMID- 15125743 TI - Intraoperative cytologic examination of breast sentinel lymph nodes: test utility and patient impact. AB - The sentinel lymph node (SLN) procedure is a method for ascertaining the axillary lymph node status in patients with breast cancer. Intraoperative examination of the SLN may be important, because a positive result directs surgery to a complete axillary lymph node dissection. Intraoperative cytologic examination (IOCE) is a method of intraoperative evaluation, although little data are available regarding the sensitivity of the method with respect to tumor size and the size of the SLN metastasis. All SLN cases for the years 1997-2002 at Magee-Womens Hospital were tabulated for primary breast carcinoma size, IOCE result, final histologic result, and size of the SLN metastasis. All SLNs had IOCE with touch imprints. Scrape SLN preparations and frozen sections were strongly discouraged. There were 748 SLN cases comprising 1576 SLNs that had IOCE, and there were 247 true positive SLN cases comprising 522 SLNs. Of the 247 true positive SLN cases, 111 had a positive IOCE (111/247; 45% sensitivity overall) and there were 136 false negatives. Of the 247 cases, 164 were SLN micrometastases < or =2.0 mm in size, and 44 (27%) of these were detected by IOCE, while the remaining 120 cases were false negative. Of the 83 SLN macrometastases (>2.0 mm), 66 (80%) were detected by IOCE, with 17 false negatives. In this series, 15 cases (2%) were given the IOCE diagnosis of atypical/defer, and all of these permanent sections were histologically positive. There were five IOCE-positive cases that were histologically negative. Of the 164 SLNs with micrometastases < or =2.0 mm, 17.6% (29/164) were < or =0.5 mm (6/29 [21.4%] were IOCE positive), 5.5% (9/164) were 0.51-1.0 mm (3/9 [33%] were IOCE positive), and 3.6% (6/164) were 1.1-2.0 mm (2/6 [33%] were IOCE positive). There were 83 SLNs with macrometastases larger than 2.0 mm, and 66/83 (80%) were detected by IOCE. In this group, 22% (18/83) were 2.1-5.0 mm (8/18 [44.4%] were IOCE positive) and 57.8% (48/83) were larger than 5.0 mm (41/48 [85%] were IOCE positive). The mean primary breast tumor size was 15.4 mm, with a mean SLN tumor size of 1.4 mm. There was a significant correlation with tumor size and the presence of SLN metastasis, and a significant correlation with tumor size and size of the SLN metastasis. There was a significant t correlation of primary tumor size and positive IOCE, with the group of negative IOCE cases having a mean tumor size of 14 mm and the positive IOCE group having a mean tumor size of 22 mm. The overall sensitivity of the method was 45%, specificity 99%, positive predictive value 0.99, and negative predictive value 0.80. Sensitivity of the IOCE procedure based on SLN tumor size is as follows: < or =0.5 mm, 21.4%; 0.51-1.0 mm, 33%; 1.1-2.0 mm, 33%; 2.1-5.0 mm, 44.4%; and >5.0 mm, 85%. Primary tumor size correlates with a positive SLN status and size of the SLN metastasis. Most false-negative IOCEs are due to micrometastases. Positive IOCE cases had a significantly larger SLN metastasis size (mean 8.0 mm) than the false-negative IOCE group (mean 1.4 mm). The IOCE of SLNs has a high negative predictive value, but this is a poor test for the detection of micrometastases, as this group accounts for the majority of false negative IOCEs of breast SLNs. PMID- 15125744 TI - Cytotoxic T lymphocyte count and survival time in women with metastatic breast cancer. AB - While prognostic factors in early stage breast cancer are well documented, few studies have examined predictors of the rate of metastatic progression. The purpose of this study was to examine cytotoxic T-cell lymphocyte (CTL) count as a marker of disease status in women with metastatic breast cancer. This study examined CTL subset counts as predictors of subsequent survival in 113 women with metastatic or recurrent breast cancer. Samples were measured by flow cytometry using monoclonal antibodies for cell surface antigens for percentages and absolute numbers of CTLs (CD3/CD8), total lymphocytes, T cells (CD3), helper T cells (CD3/CD4), and total white blood cell (TWC) count. Higher CTL counts emerged as a significant predictor of longer survival up to 7 years later (Wald = 7.40, p = 0.007; Cox regression model). The relationship of higher CTL count with enhanced survival was independent of the effects of medical treatment. CTLs were significantly associated with TWC count (r = 0.42, p < 0.001). However, TWC count was not associated with subsequent survival time. Higher CTL count was associated with Karnofsky performance status (r = 0.27, p = 0.004). However, after adjustment for the Karnofsky score, the CTL count/survival relationship remained significant (Wald = 4.33, p = 0.038). In conclusion, there is a robust relationship between CTL count and survival that is independent of the effects of medical treatments, TWC count, and Karnofsky performance status. Moreover, a reduced CTL count may be a mediator or marker of more rapid disease progression in metastatic breast cancer. PMID- 15125745 TI - Microtome shaving radiography: demonstration of loss of mammographic microcalcifications during histologic sectioning. AB - Mammographic-pathologic correlation of suspicious microcalcifications is essential for optimal diagnosis and local staging of early breast carcinoma. Loss of microcalcifications during histologic sectioning has been suggested as one reason for the occasional lack of microscopic visualization of microcalcifications in routinely processed breast biopsy specimens obtained for suspicious mammographic microcalcifications. Two case reports utilizing radiography of histologic shavings of stereotactic core biopsies and surgical excisional biopsies of mammographic microcalcifications provide concrete evidence of the loss of large calcific particles during the microtome process. PMID- 15125746 TI - A phase I dose escalation trial of gemcitabine with radiotherapy for breast cancer in the treatment of unresectable chest wall recurrences. AB - The purpose of this study was to determine the maximum tolerated dose (MTD) of gemcitabine when given concurrently with standard radiotherapy for the treatment of chest wall recurrences, and to compare actuarial rates of local-regional control with those achieved in historical controls. Patients with unresectable chest wall recurrences were enrolled in a phase I trial of concurrent gemcitabine and radiotherapy. Gemcitabine was increased at 150 mg/m(2)/week increments, starting at 300 mg/m(2)/week. Radiotherapy was delivered to the chest wall and regional nodes to a total of 60 to 70 Gy in 2 Gy daily fractions. Treatment toxicity was assessed and a comparison of treatment outcome was performed between study patients and historical groups treated with either radiotherapy alone or excision followed by radiotherapy. The dose-limiting toxicities of neutropenia and thrombocytopenia occurred at the second planned dose of 450 mg/m(2)/week after accrual of only six patients, resulting in a MTD of 300 mg/m(2)/week. Myelosuppression and skin desquamation were commonly observed. Actuarial rates of local-regional control were 100%, 50%, and 90% at 2 years for the gemcitabine with radiotherapy, radiotherapy alone, and excision followed by radiotherapy groups, respectively (p = 0.105). The difference among the Kaplan-Meier curves for overall local-regional control was statistically significant at p = 0.007 in favor of combined gemcitabine and radiotherapy. The MTD of gemcitabine is 300 mg/m(2)/week when gemcitabine is delivered concurrently with radiotherapy for unresectable chest wall failures. This novel approach suggests excellent local regional control when compared to historical controls. A phase II trial is warranted. PMID- 15125747 TI - The impact of hormone receptor status on the clinical efficacy of the new generation aromatase inhibitors: a review of data from first-line metastatic disease trials in postmenopausal women. AB - Hormone receptor status has long been considered important in predicting the efficacy of endocrine agents for the treatment of breast cancer. We aim to address whether hormone receptor status influences treatment outcome in postmenopausal women receiving aromatase inhibitors for advanced breast cancer. We include data from three phase III clinical trials, comparing the activity of the new-generation aromatase inhibitors, anastrozole or letrozole, with tamoxifen as a first-line treatment. For both agents, a significant relationship was observed between hormone receptor status and time to disease progression (TTP), with increased TTP seen in patients with a higher confirmed percentage of estrogen receptor (ER)- and/or progesterone receptor (PR)-positive tumors. A relationship between objective response rate (complete or partial response) or clinical benefit (complete or partial response or stabilization for > or =24 weeks) and hormone receptor status was apparent for anastrozole but not letrozole treatment. Overall these data confirm that hormone receptor status should be a strong consideration in the selection of endocrine treatment for postmenopausal women with advanced breast cancer. PMID- 15125748 TI - Prognosis of breast cancer patients with familial history classified according to their menopausal status. AB - Breast cancer patients were classified in the family history positive (FHP) group when they had at least one second-degree relative who was a breast cancer patient. The results of a comparative study with patients classified in the family history negative (FHN) group showed the prognosis of the FHP group was significantly better than that of the FHN group. However, when those patients were classified according to their menopausal status at onset, there were no significant differences in survival rates between the FHP and FHN groups with onset before menopause, whereas the survival rate of the FHP group was significantly higher than that of the FHN group with onset after menopause. The same results were found when the FHP group was subgrouped into the FHP group with first-degree relatives and the FHP group with second-degree relatives. Further investigations on background factors revealed that the patients with onset before menopause showed no significant differences between the FHP and FHN groups in age at surgery, diameter of the tumor, histologic grade, the number of metastatic lymph nodes, body weight, estrogen receptor (ER) status, and the values of CEA and CA15-3 before surgery. On the other hand, the FHP patients with onset after menopause showed significantly lower numbers of metastatic lymph nodes and trends showing higher ER values and lower CA15-3-values. Therefore the favorable prognosis in the FHP group seems to be attributable to the higher survival rate of the FHP patients with onset after menopause. PMID- 15125749 TI - Quality of life following breast-conserving therapy or mastectomy: results of a 5 year prospective study. AB - There are many conflicting results in the literature comparing quality of life following breast-conserving therapy (BCT) and mastectomy. This study compared long-term quality of life between breast cancer patients treated by BCT or mastectomy in three age groups. Patients (n = 990) completed a quality of life survey, including the European Organization for Research and Treatment of Cancer Quality of Life Core Questionnaire (EORTC QLQ-C30), at regular intervals over 5 years. In the cross-sectional data, mastectomy patients had significantly (p < 0.01) lower body image, role, and sexual functioning scores and their lives were more disrupted than BCT patients. Emotional and social functioning and financial and future health worries were significantly (p < 0.01) worse for younger patients. There were no differences in body image and lifestyle scores between age groups. There was also no interaction between age and surgery method. Even patients > or =70 years of age reported higher body image and lifestyle scores when treated with BCT. The repeated measures analysis indicated that four functioning scores, half the symptom scores, future health, and global quality of life improved significantly (p < 0.01) over time. All these variables increased significantly for BCT patients and those 50 to 69 years of age. Body image, sexual functioning, and lifestyle disruption scores did not improve over time. BCT should be encouraged in all age groups. Coping with appearance change should be addressed in patient interventions. PMID- 15125750 TI - Magnetic resonance imaging of benign phyllodes tumors of the breast. AB - Magnetic resonance imaging (MRI) has the potential to become a useful adjunct in breast imaging. Contrast-enhanced breast MRI has demonstrated a high sensitivity in the detection of benign and malignant breast disease. Our study aimed to correlate the dynamic contrast-enhanced MRI appearance of benign phyllodes tumor of the breast with histopathologic findings. We retrospectively reviewed the MRI findings in eight patients with benign phyllodes tumor of the breast to describe the image characteristics of this disease. The architectural features and enhancement patterns of this tumor were assessed and compared with other breast diseases. MRIs demonstrated some characteristics for large benign phyllodes tumors (more than 3 cm in size). On T(2)-weighted images, they were imaged as spotted tumors in high to iso signal intensity with cystic components or septations inside. In the time-signal intensity curve for the eight patients in our study who underwent dynamic MRI, we demonstrated two patterns of their curve: rapidly and gradually enhanced. In conclusion, MRI findings in benign phyllodes tumor include dynamic curves of gradually and rapidly enhancing types, and a low and inhomogeneous signal intensity on T(2)-weighted images compared with fibroadenoma. These findings appear to be useful for diagnosis. PMID- 15125751 TI - Male mammary fistula complicating senescent gynecomastia. AB - Mammary fistulas occurring in males are extremely rare, the first being described in 1974 by Tedeschi et al. (1). Our detailed review of the world literature has revealed a total of just 20 cases of mammary fistulas in males; a total which includes three cases of mammary fistulas in infants and three cases associated with human immunodeficiency virus (HIV) infection. Mammary fistulas develop between the lactiferous ducts and areolar skin. Altered duct anatomy and microarchitecture may predispose a patient to mammary fistulas. This discussion focuses on mammary fistulas in males where smoking is a likely etiologic factor, and therefore examines a clinical rarity with just 14 reported cases in the literature. The treatment of mammary fistulas in males is the same as for females once the diagnosis has been made. PMID- 15125752 TI - Laser therapy of breast cancer with 3-year follow-up. AB - Breast cancer is the commonest female malignancy in the United States and is increasingly being detected at a non-palpable stage by annual mammography. Minimally invasive methods of its local treatment have been recently introduced as alternative to its surgical removal. The case reported in this article illustrates the advantages of in-situ ablation with minimal discomfort to the patient and deformity of the breast. More importantly, it demonstrates the absence of any adverse effect on health and survival of the patient during this intermediate period of follow-up. PMID- 15125753 TI - Efavirenz-associated gynecomastia: report of five cases and review of the literature. AB - The prognosis of HIV infection has improved dramatically since the introduction of highly active antiretroviral therapy (HAART). However, numerous adverse effects and limitations regarding tolerability remain a concern. Lipomastia (pseudogynecomastia), a breast enlargement due to central adiposity, may occur as part of a fat redistribution syndrome which has been associated with HAART regimens and several pathogenic mechanisms have been advocated in its development. Here we report an observational longitudinal study of five patients diagnosed of gynecomastia associated with efavirenz-based HAART regimens. All cases reached successful immunologic and virologic responses to HAART. The delay of appearance of gynecomastia from the beginning of HAART ranged between 4 to 15 months. In all five cases, gynecomastia regressed after efavirenz withdrawal (mean period of 5 months). In summary, we think that HAART induced gynecomastia should be suspected in HIV patients receiving efavirenz-containing regimens. Although pathogenesis is unclear, this study and a review of the English literature implicates two possible mechanisms: (a) immune restoration processes and (b) efavirenz mediated estradiol-like effects. PMID- 15125754 TI - Postlactational microcalcifications. AB - A case of postlactational microcalcifications is reported. A 42-year-old woman presented for screening mammography 2 months after completion of breast-feeding. Comparison to her pregravid screening mammogram revealed the appearance of multiple groups of indeterminate microcalcifications bilaterally (BIRADS IV). She underwent bilateral stereotactic core biopsies of representative areas, yielding benign pathology. There have been anecdotal accounts and five reported cases of lactational microcalcifications in the radiology literature. We discuss the possible etiologies as well as implications of this mammographic finding. PMID- 15125755 TI - Mondor's disease of the axilla: a rare complication of sentinel node biopsy. AB - Three cases of Mondor's disease of the axilla following sentinel lymph node biopsy (SLNB) are described. In all cases we used the combination of blue dye and radiocolloid, and complete axillary dissection was not performed. The numbers of lymph nodes removed in each case were five, four, and two, respectively. All the events of Mondor's disease resolved spontaneously or following a short therapy of anti-inflammatory agents. PMID- 15125756 TI - Increase in mammographic breast density associated with the use of domperidone. PMID- 15125757 TI - Myolipoma of the breast: mammographic, sonographic, and pathologic correlation. PMID- 15125758 TI - Granulomatous lobular mastitis misdiagnosed as breast carcinoma. PMID- 15125759 TI - Ductal carcinoma in situ (DCIS) of the male breast. PMID- 15125760 TI - Male breast: apocrine ductal papilloma with psammoma bodies. PMID- 15125761 TI - Relationship of ethnicity and histologic type of breast carcinoma in a West Indian population. PMID- 15125762 TI - Comment on Rahusen et al. PMID- 15125764 TI - Silencing of subfamily I of protein phosphatase 2A catalytic subunits results in activation of plant defense responses and localized cell death. AB - The central importance of protein phosphorylation in plant defense responses has been demonstrated by the isolation of several disease-resistance genes that encode protein kinases. In addition, there are many reports of changes in protein phosphorylation accompanying plant responses to pathogens. In contrast, little is known about the role of protein dephosphorylation in regulating plant defenses. We report that expression of the LePP2Ac1 gene, which encodes a catalytic subunit of the heterotrimeric protein phosphatase 2A (PP2Ac), is rapidly induced in resistant tomato leaves upon inoculation with an avirulent strain of Pseudomonas syringae pv. tomato. By analysis of PP2Ac gene sequences from several plant species, we found that PP2Ac genes cluster into two subfamilies, with LePP2Ac1 belonging to subfamily I. Virus-induced gene silencing (VIGS) in Nicotiana benthamiana was used to suppress expression of genes from subfamily I and not from subfamily II. The PP2Ac-silenced plants had greatly decreased PP2A activity, constitutively expressed pathogenesis-related (PR) genes, and developed localized cell death in stems and leaves. In addition, the plants were more resistant to a virulent strain of P. syringae pv. tabaci and showed an accelerated hypersensitive response (HR) to effector proteins from both P. syringae and the fungal pathogen, Cladosporium fulvum. Thus, catalytic subunits of PP2Ac subfamily I act as negative regulators of plant defense responses likely by de-sensitizing protein phosphorylation cascades. PMID- 15125766 TI - Spreading of post-transcriptional gene silencing along the target gene promotes systemic silencing. AB - Transitive silencing and grafting-induced gene silencing phenomena were combined to investigate whether a primary target beta-glucuronidase (gus) gene could promote the generation of systemic transitive silencing signals. Tobacco plants with hemizygous or homozygous silencer locus and in trans silenced primary target were used as a source of post-transcriptionally silenced rootstocks and tobacco plants with or without a secondary target locus as scion source. The silencer locus harbored two identical neomycin phosphotransferase II (nptII)-containing T DNAs, integrated as an inverted repeat. The primary target locus carried a gus gene with homology to the transcribed region of the nptII gene only in the 3' untranslated region, whereas the secondary target locus had two or more copies of a gus gene without homology to transcribed sequences of the silencer locus. The upstream region of the initially targeted sequences of the in trans silenced gus gene could induce the production of a systemic signal. This signal was capable of triggering post-transcriptional gene silencing (PTGS) of the secondary target gus genes in the scion. In addition, the induction of systemic silencing was strikingly dosage dependent for the silencer as well as the primary target loci in the rootstock. Moreover, in the scions, the secondary target gus genes had to be present to generate detectable amounts of short interfering RNAs. PMID- 15125765 TI - Cytokinin stimulates polyribosome loading of nuclear-encoded mRNAs for the plastid ATP synthase in etioplasts of Lupinus luteus: the complex accumulates in the inner-envelope membrane with the CF(1) moiety located towards the stromal space. AB - Three of the nine subunits of the plastid ATP synthase, including the subunit of the CF(1) moiety (gene AtpC), are encoded in the nucleus. Application of cytokinin to etiolated lupine seedlings induces polyribosome association of their mRNAs. This appears to be specific as no such regulation was observed for messages for three ribosomal proteins. Cytokinin-mediated polyribosome loading was also observed for the spinach AtpC message in etiolated transgenic tobacco seedlings. Analysis of various spinach AtpC mRNA derivatives uncovered that the 5' untranslated region (5' UTR) of this message is sufficient to direct polyribosome loading, and that sequences at the 3' end of the AtpC 5' UTR, including an UC-rich motif, are crucial for this regulation. The increase in polyribosome loading of the AtpC message correlated with an increased synthesis of the polypeptide. The subunit, together with the ATP synthase complex, accumulates in the inner-envelope membrane with the CF(1) moiety located towards the stromal space of the etioplast. These results suggest that cytokinin promotes accumulation of the ATP synthase in the inner-envelope membrane of lupine etioplasts by stimulating the translation efficiency of their nuclear-encoded messages. PMID- 15125767 TI - Differential stage-specific regulation of cyclin-dependent kinases during cambial dormancy in hybrid aspen. AB - The cambium of woody plants cycles between active and dormant states. Dormancy can be subdivided into eco- and endodormant stages. Ecodormant trees resume growth upon exposure to growth-promotive signals, while the establishment of endodormant state results in loss of the ability to respond to these signals. In this paper, we analysed the regulation of cyclin-dependent kinases (CDKs) to understand the differential response of cell division machinery to growth promotive signals during the distinct stages of dormancy in hybrid aspen. We show that 4 weeks of short-day (SD) treatment causes termination of the cambial cell division and establishment of the ecodormant state. This coincides with a steady decline in the histone H1 kinase activity of the PSTAIRE-type poplar CDKA (PttCDKA) and the PPTTLRE-type PttCDKB kinase complexes. However, neither the transcript nor the polypeptide levels of PttCDKA and PttCDKB are reduced during ecodormancy. In contrast, 6 weeks of SD treatment establishes endodormancy, which is marked by the reduction and disappearance of the PttCDKA and PttCDKB protein levels and the PttCDKB transcript levels. The transition to endodormancy is preceded by an elevated E2F (adenosine E2 promoter binding factor) phosphorylation activity of the PttCDKA kinase that reduces the DNA-binding activity of E2F in vitro. The transition to endodormancy is followed by a reduction of retinoblastoma (Rb) phosphorylation activity of PttCDKA protein complexes. Both phosphorylation events could contribute to block the G1 to S phase transition upon the establishment of endodormancy. Our results indicate that eco- and endodormant stages of cambial dormancy involve a stage-specific regulation of the cell cycle effectors at multiple levels. PMID- 15125768 TI - Differential stability of Arabidopsis D-type cyclins: CYCD3;1 is a highly unstable protein degraded by a proteasome-dependent mechanism. AB - In Arabidopsis, the D-type cyclin CYCD3 is rate-limiting for transition of the G(1)/S boundary, and is transcriptionally upregulated at this point in cells re entering the cell cycle in response to plant hormones and sucrose. However, little is known about the regulation of plant cell-cycle regulators at the protein level. We show here that CYCD3;1 is a phosphoprotein highly regulated at the level of protein abundance, whereas another D-type cyclin CYCD2;1 is not. The level of CYCD3;1 protein falls rapidly on sucrose depletion, correlated with the arrest of cells in G(1) phase, suggesting a rapid turnover of CYCD3;1. Treatment of exponentially growing cells with the protein synthesis inhibitor cycloheximide (CHX) confirms that CYCD3;1 is normally a highly unstable protein, with a half life of approximately 7 min on CHX treatment. In both sucrose-starved and exponentially growing cells, CYCD3;1 protein abundance increases in response to treatment with MG132 (carbobenzoxyl-leucinyl-leucinyl-leucinal), a reversible proteasome inhibitor, but not in response to the cysteine protease inhibitor E-64 or the calpain inhibitor ALLN (N-acetyl-leucyl-leucyl-norleucinal). The increase on MG132 treatment is because of de novo protein synthesis coupled with the blocking of CYCD3;1 degradation. Longer MG132 treatment leads to C-terminal cleavage of CYCD3;1, accumulation of a hyperphosphorylated form and its subsequent disappearance. We conclude that CYCD3;1 is a highly unstable protein whose proteolysis is mediated by a proteasome-dependent pathway, and whose levels are highly dependent on the rate of CYCD3;1 protein synthesis. PMID- 15125769 TI - A recessive mutation in the RUB1-conjugating enzyme, RCE1, reveals a requirement for RUB modification for control of ethylene biosynthesis and proper induction of basic chitinase and PDF1.2 in Arabidopsis. AB - By screening etiolated Arabidopsis seedlings for mutants with aberrant ethylene related phenotypes, we identified a mutant that displays features of the ethylene mediated triple response even in the absence of ethylene. Further characterization showed that the phenotype observed for the dark-grown seedlings of this mutant is reversible by prevention of ethylene perception and is dependent on a modest increase in ethylene production correlated with an increase in 1-aminocyclopropane-1-carboxylic acid (ACC) oxidase (ACO) activity in the hypocotyl. Molecular characterization of leaves of the mutant revealed severely impaired induction of basic chitinase (chiB) and plant defensin (PDF)1.2 following treatment with jasmonic acid and/or ethylene. Positional cloning of the mutation resulted in identification of a 49-bp deletion in RCE1 (related to ubiquitin 1 (RUB1)-conjugating enzyme), which has been demonstrated to be responsible for covalent attachment of RUB1 to the SCF (Skpl Cdc 53 F-box) ubiquitin ligase complex to modify its activity. Our analyses with rce1-2 demonstrate a previously unknown requirement for RUB1 modification for regulation of ethylene biosynthesis and proper induction of defense-related genes in Arabidopsis. PMID- 15125770 TI - Herbivore-induced plant vaccination. Part I. The orchestration of plant defenses in nature and their fitness consequences in the wild tobacco Nicotiana attenuata. AB - A plant's responses to attack from particular pathogens and herbivores may result in resistance to subsequent attack from the same species, but may also affect different species. Such a cross-resistance, called immunization or vaccination, can benefit the plant, if the fitness consequences of attack from the initial attacker are less than those from subsequent attackers. Here, we report an example of naturally occurring vaccination of the native tobacco plant, Nicotiana attenuata, against Manduca hornworms by prior attack from the mirid bug, Tupiocoris notatus (Dicyphus minimus), which results from the elicitation of two categories of induced plant responses. First, attack from both herbivore species causes the plants in nature to release predator-attracting volatile organic compounds (VOCs), and the attracted generalist predator, Geocoris pallens, preferentially attacks the less mobile hornworm larvae. Second, attack from both mirids and hornworms increases the accumulation of secondary metabolites and proteinase inhibitors (PIs) in the leaf tissue, which is correlated with the slow growth of Manduca larvae. Mirid damage does not significantly reduce the fitness of the plant in nature, whereas attack from the hornworm reduces lifetime seed production. Consequently, plants that are attacked by mirids realize a significant fitness advantage in environments with both herbivores. The combination of growth-slowing direct defenses and predator-attracting indirect defenses results in greater hornworm mortality on mirid-attacked plants and provides the mechanism of the vaccination phenomenon. PMID- 15125771 TI - Herbivore-induced plant vaccination. Part II. Array-studies reveal the transience of herbivore-specific transcriptional imprints and a distinct imprint from stress combinations. AB - Summary Microarray technology has given plant biologists the ability to simultaneously monitor changes in the expression of hundreds of genes, and yet, to date, this technology has not been applied to ecological phenomena. In native tobacco (Nicotiana attenuata), prior attack of sap-feeding mirids (Tupiocoris notatus) results in vaccination of the plant against subsequent attacks by chewing hornworms (Manduca sexta). This vaccination is mediated by a combination of direct and indirect defenses and tolerance responses, which act in concert with the attack preferences of a generalist predator. Here, we use microarrays enriched in herbivore-elicited genes with a principal components analysis (PCA) to characterize transcriptional 'imprints' of single, sequential, or simultaneous attacks by these two main herbivores of N. attenuata. The PCA identified distinctly different imprints left by individual attack from the two species after 24 h, but not after 5 days. Moreover, imprints of sequential or simultaneous attacks differed significantly from those of single attack, suggesting the existence of a distinct gene expression program responsive to the combination of biological stressors. A dissection of the transcriptional imprints revealed responses in direct and indirect defense genes that were well correlated with observed increases in defense metabolites. Attack from both herbivores elicits a switch from growth- to defense-related transcriptional processes, and herbivore-specific changes occur largely in primary metabolism and signaling cascades. PCA of these polygenic transcriptional imprints characterizes the ephemeral changes in the transcriptome that occur during the maturation of ecologically relevant phenotypic responses. PMID- 15125772 TI - EARLY FLOWERING 5 acts as a floral repressor in Arabidopsis. AB - EARLY FLOWERING 5 (ELF5) is a single-copy gene involved in flowering time regulation in Arabidopsis. ELF5 encodes a nuclear-targeted protein that is related to the human nuclear protein containing a WW domain (Npw)38-binding protein (NpwBP). Lesions in ELF5 cause early flowering in both long days and short days. elf5 mutations partially suppress the late flowering of both autonomous-pathway mutants and FRIGIDA (FRI)-containing lines by reducing the expression of FLOWERING LOCUS C (FLC), a floral repressor upon which many of the flowering pathways converge. elf5 mutations also partially suppress photoperiod pathway mutants, and this, along with the ability of elf5 mutations to cause early flowering in short days, indicates that ELF5 also affects flowering independently of FLC. PMID- 15125773 TI - A novel Arabidopsis gene TONSOKU is required for proper cell arrangement in root and shoot apical meristems. AB - Root apical meristem (RAM) and shoot apical meristem (SAM) are vital for the correct development of the plant. The direction, frequency, and timing of cell division must be tightly controlled in meristems. Here, we isolated new Arabidopsis mutants with shorter roots and fasciated stems. In the tonsoku (tsk) mutant, disorganized RAM and SAM formation resulted from the frequent loss of proper alignment of the cell division plane. Irregular cell division also occurred in the tsk embryo, and the size of cells in meristems and embryo in tsk mutant was larger than in the wild type. In the enlarged SAM of the tsk mutant, multiple centers of cells expressing WUSCHEL (WUS) were observed. In addition, expression of SCARECROW (SCR) in the quiescent center (QC) disappeared in the disorganized RAM of tsk mutant. These results suggest that disorganized cell arrangements in the tsk mutants result in disturbed positional information required for the determination of cell identity. The TSK gene was found to encode a protein with 1311 amino acids that possesses two types of protein-protein interaction motif, leucine-glycine-asparagine (LGN) repeats and leucine-rich repeats (LRRs). LGN repeats are present in animal proteins involved in asymmetric cell division, suggesting the possible involvement of TSK in cytokinesis. On the other hand, the localization of the TSK-GFP (green fluorescent protein) fusion protein in nuclei of tobacco BY-2 cells and phenotypic similarity of tsk mutants to other fasciated mutants suggest that the tsk mutation may cause disorganized cell arrangements through defects in genome maintenance. PMID- 15125774 TI - Sorting signals in the cytosolic tail of membrane proteins involved in the interaction with plant ARF1 and coatomer. AB - In mammals and yeast, a cytosolic dilysine motif is critical for endoplasmic reticulum (ER) localization of type I membrane proteins. Retrograde transport of type I membrane proteins containing dilysine motifs at their cytoplasmic carboxy (C)-terminal tail involves the interaction of these motifs with the COPI coat. The C-terminal dilysine motif has also been shown to confer ER localization to type I membrane proteins in plant cells. Using in vitro binding assays, we have analyzed sorting motifs in the cytosolic tail of membrane proteins, which may be involved in the interaction with components of the COPI coat in plant cells. We show that a dilysine motif in the -3,-4 position (relative to the cytosolic C terminus) recruits in a very specific manner all the subunits of the plant coatomer complex. Lysines cannot be replaced by arginines or histidines to bind plant coatomer. A diphenylalanine motif in the -7,-8 position, which by itself has a low ability to bind plant coatomer, shows a clear cooperativity with the dilysine motif. Both dilysine and diphenylalanine motifs are present in the cytosolic tail of several proteins of the p24 family of putative cargo receptors, which has several members in plant cells. The cytosolic tail of a plant p24 protein is shown to recruit not only coatomer but also ADP ribosylation factor 1 (ARF1), a process which depends on both dilysine and diphenylalanine motifs. ARF1 binding increases twofold upon treatment with brefeldin A (BFA) and is completely abolished upon treatment with GTPgammaS, suggesting that ARF1 can only interact with the cytosolic tail of p24 proteins in its GDP-bound form. PMID- 15125775 TI - Overexpression of a novel small peptide ROTUNDIFOLIA4 decreases cell proliferation and alters leaf shape in Arabidopsis thaliana. AB - Leaf shape is determined by polar cell expansion and polar cell proliferation along the leaf axes. However, the genes controlling polar cell proliferation during leaf morphogenesis are largely unknown. We identified a dominant mutant of Arabidopsis thaliana, rotundifolia4-1D (rot4-1D), which possessed short leaves and floral organs. We showed that the altered leaf shape is caused by reduced cell proliferation, specifically in the longitudinal (proximal-distal) axis of the leaf, suggesting that the ROT4 gene controls polar cell proliferation in lateral organs. The ROT4 open-reading frame (ORF) encodes a novel small peptide that had not been identified in the Arabidopsis genome annotation. Overexpression of a ROT4-green fluorescence protein (GFP) fusion protein in transgenic plants recapitulated the rot4 phenotype, suggesting that ROT4 acts to restrict cell proliferation. The ROT4-GFP fusion protein localized to the plasma membrane when expressed in transgenic Arabidopsis plants. Phylogenetic analysis indicates that ROT4 defines a novel seed plant-specific family of small peptides with 22 members in Arabidopsis, ROT FOUR LIKE1-22 (RTFL1-22). All RTFL members share a conserved 29-amino acid domain, the RTF domain, and overexpression of the ROT4 RTF domain alone is sufficient to confer a rot4-1D phenotype. Loss-of-function mutations in several RTFL genes were aphenotypic, suggesting that there may be some functional redundancy between family members. Analyses by reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization revealed that ROT4 is expressed in the shoot apex and young leaves of wild-type plants, consistent with a role for ROT4 in controlling polarity-dependent cell proliferation during wild-type leaf morphogenesis. PMID- 15125776 TI - Membrane transport of camptothecin: facilitation by human P-glycoprotein (ABCB1) and multidrug resistance protein 2 (ABCC2). AB - BACKGROUND: The purpose of the present study was to continue the investigation of the membrane transport mechanisms of 20-(S)-camptothecin (CPT) in order to understand the possible role of membrane transporters on its oral bioavailability and disposition. METHODS: The intestinal transport kinetics of CPT were characterized using Caco-2 cells, MDCKII wild-type cells and MDCKII cells transfected with human P-glycoprotein (PGP) (ABCB1) or human multidrug resistance protein 2 (MRP2) (ABCC2). The effects of drug concentration, inhibitors and temperature on CPT directional permeability were determined. RESULTS: The absorptive (apical to basolateral) and secretory (basolateral to apical) permeabilities of CPT were found to be saturable. Reduced secretory CPT permeabilities with decreasing temperatures suggests the involvement of an active, transporter-mediated secretory pathway. In the presence of etoposide, the CPT secretory permeability decreased 25.6%. However, inhibition was greater in the presence of PGP and of the breast cancer resistant protein inhibitor, GF120918 (52.5%). The involvement of additional secretory transporters was suggested since the basolateral to apical permeability of CPT was not further reduced in the presence of increasing concentrations of GF120918. To investigate the involvement of specific apically-located secretory membrane transporters, CPT transport studies were conducted using MDCKII/PGP cells and MDCKII/MRP2 cells. CPT carrier-mediated permeability was approximately twofold greater in MDCKII/PGP cells and MDCKII/MRP2 cells than in MDCKII/wild-type cells, while the apparent Km values were comparable in all three cell lines. The efflux ratio of CPT in MDCKII/PGP in the presence of 0.2 microM GF120918 was not completely reversed (3.36 to 1.49). However, the decrease in the efflux ratio of CPT in MDCKII/MRP2 cells (2.31 to 1.03) suggests that CPT efflux was completely inhibited by MK571, a potent inhibitor of the Multidrug Resistance Protein transporter family. CONCLUSIONS: The current results provide evidence that PGP and MRP2 mediate the secretory transport of CPT in vitro. However, the involvement of other transporters cannot be ruled out based on these studies. Since these transporters are expressed in the intestine, liver and kidney variations in their expression levels and/or regulation may be responsible for the erratic oral absorption and biliary excretion of CPT observed in human subjects. PMID- 15125777 TI - CHK2 kinase expression is down-regulated due to promoter methylation in non-small cell lung cancer. AB - BACKGROUND: CHK2 kinase is a tumor suppressor that plays important role in DNA damage signaling, cell cycle regulation and DNA damage induced apoptosis. CHK2 kinase expression was known to be ubiquitous in mammalian cells. CHK2-/- cells were remarkably resistant to DNA damage induced apoptosis, mimicking the clinical behavior of non-small cell lung cancer to conventional chemo and radiation therapy. RESULT: We reported that the CHK2 expression is diminished or absent in both non-small cell lung cancer (NSCLC) cell lines and clinical lung cancer tumor specimens. The absent CHK2 expression in NSCLC was due to hypermethylation of the CHK2 gene promoter, preventing from binding of a transcriptional factor, leading to silence of the CHK2 gene transcription. CONCLUSION: Since the CHK2 null mice showed a remarkable radioresistance, which bear significant similarity to clinical behavior of NSCLC, down-regulation of CHK2 kinase expression by CHK2 gene silencing and methylation in non-small cell lung cancer suggest a critical role of CHK2 kinase in DNA damage induced apoptosis and a novel mechanism of the resistance of NSCLC to DNA damage based therapy. PMID- 15125778 TI - Characterization of potential larval habitats for Anopheles mosquitoes in relation to urban land-use in Malindi, Kenya. AB - BACKGROUND: This study characterized Anopheles mosquito larval habitats in relation to ecological attributes about the habitat and community-level drainage potential, and investigated whether agricultural activities within or around urban households increased the probability of water body occurrence. Malindi, a city on the coast of Kenya, was mapped using global positioning system (GPS) technology, and a geographic information system (GIS) was used to overlay a measured grid, which served as a sampling frame. Grid cells were stratified according to the level of drainage in the area, and 50 cells were randomly selected for the study. Cross-sectional household and entomological surveys were conducted during November and December 2002 within the 50 grid cells. Chi-square analysis was used to test whether water bodies differed fundamentally between well and poorly drained areas, and multi-level logistic regression was used to test whether household-level agricultural activity increased the probability of water body occurrence in the grid cell. RESULTS: Interviews were conducted with one adult in 629 households. A total of 29 water bodies were identified within the sampled areas. This study found that characteristics of water bodies were fundamentally the same in well and poorly drained areas. This study also demonstrated that household-level urban agriculture was not associated with the occurrence of water bodies in the grid cell, after controlling for potential confounders associated with distance to the city center, drainage, access to resources, and population density. CONCLUSIONS: Household-level urban agricultural activity may be less important than the other types of human perturbation in terms of mosquito larval habitat creation. The fact that many larvae were coming from few sites, and few sites in general were found under relatively dry conditions suggests that mosquito habitat reduction is a reasonable and attainable goal in Malindi. PMID- 15125779 TI - Physiological basis and image processing in functional magnetic resonance imaging: neuronal and motor activity in brain. AB - Functional magnetic resonance imaging (fMRI) is recently developing as imaging modality used for mapping hemodynamics of neuronal and motor event related tissue blood oxygen level dependence (BOLD) in terms of brain activation. Image processing is performed by segmentation and registration methods. Segmentation algorithms provide brain surface-based analysis, automated anatomical labeling of cortical fields in magnetic resonance data sets based on oxygen metabolic state. Registration algorithms provide geometric features using two or more imaging modalities to assure clinically useful neuronal and motor information of brain activation. This review article summarizes the physiological basis of fMRI signal, its origin, contrast enhancement, physical factors, anatomical labeling by segmentation, registration approaches with examples of visual and motor activity in brain. Latest developments are reviewed for clinical applications of fMRI along with other different neurophysiological and imaging modalities. PMID- 15125780 TI - Formulation of photon diffusion from spherical bioluminescent sources in an infinite homogeneous medium. AB - BACKGROUND: The bioluminescent enzyme firefly luciferase (Luc) or variants of green fluorescent protein (GFP) in transformed cells can be effectively used to reveal molecular and cellular features of neoplasia in vivo. Tumor cell growth and regression in response to various therapies can be evaluated by using bioluminescent imaging. In bioluminescent imaging, light propagates in highly scattering tissue, and the diffusion approximation is sufficiently accurate to predict the imaging signal around the biological tissue. The numerical solutions to the diffusion equation take large amounts of computational time, and the studies for its analytic solutions have attracted more attention in biomedical engineering applications. METHODS: Biological tissue is a turbid medium that both scatters and absorbs photons. An accurate model for the propagation of photons through tissue can be adopted from transport theory, and its diffusion approximation is applied to predict the imaging signal around the biological tissue. The solution to the diffusion equation is formulated by the convolution between its Green's function and source term. The formulation of photon diffusion from spherical bioluminescent sources in an infinite homogeneous medium can be obtained to accelerate the forward simulation of bioluminescent phenomena. RESULTS: The closed form solutions have been derived for the time-dependent diffusion equation and the steady-state diffusion equation with solid and hollow spherical sources in a homogeneous medium, respectively. Meanwhile, the relationship between solutions with a solid sphere source and ones with a surface sphere source is obtained. CONCLUSION: We have formulated solutions for the diffusion equation with solid and hollow spherical sources in an infinite homogeneous medium. These solutions have been verified by Monte Carlo simulation for use in biomedical optical imaging studies. The closed form solution is highly accurate and more computationally efficient in biomedical engineering applications. By using our analytic solutions for spherical sources, we can better predict bioluminescent signals and better understand both the potential for, and the limitations of, bioluminescent tomography in an idealized case. The formulas are particularly valuable for furthering the development of bioluminescent tomography. PMID- 15125781 TI - Low literacy Chinese patients: how are they affected and how do they cope with health matters? A qualitative study. AB - BACKGROUND: Low literacy is common in Hong Kong and China, especially among the females and the elderly. These are often the patients with the greatest health care needs. However little is known about how low literacy affects the ability of patients to obtain adequate medical care and comply with the recommendations of health professionals. METHODS: Qualitative study of subjects' opinions using semi structured group and individual interviews. Thirty-six participants took part in 6 group and 9 individual interviews. RESULTS: Participants' opinions towards low literacy were explored in the interviews. They expressed diverse views on how they perceived low literacy. Some thought they were handicapped by it but others felt it was not a problem. Age, gender, chronic illnesses and their attitudes towards self were found to be major factors affecting their perception. Despite their low literacy, some had their specific ways to overcome the difficulties, for example, drawing symbols to help with the timing of taking the medications. Most of them felt better explanation by the health care workers was most important. CONCLUSIONS: It is highly relevant for health care workers to be aware of the low literacy status of their patients and to explain information so that patients understand it. PMID- 15125782 TI - Reversal of end-stage renal disease after aortic dissection using renal artery stent: a case report. AB - BACKGROUND: Medical management is the conventional treatment for Stanford Type B aortic dissections as surgery is associated with significant morbidity and mortality. The advent of endovascular interventional techniques has revived interest in treating end-organ complications of Type B aortic dissection. We describe a patient who benefited from endovascular repair of renal artery stenosis caused by a dissection flap, which resulted in reversal of his end-stage renal disease (ESRD). CASE PRESENTATION: A 69 y/o male with a Type B aortic dissection diagnosed two months earlier was found to have a serum creatinine of 15.2 mg/dL (1343.7 micromol/L) on routine visit to his primary care physician. An MRA demonstrated a rightward spiraling aortic dissection flap involving the origins of the celiac artery, superior mesenteric artery, and both renal arteries. The right renal artery arose from the false lumen with lack of blood flow to the right kidney. The left renal artery arose from the true lumen, but an intimal dissection flap appeared to be causing an intermittent stenosis of the left renal artery with compromised blood flow to the left kidney. Endovascular reconstruction with of the left renal artery with stent placement was performed. Hemodialysis was successfully discontinued six weeks after stent placement. CONCLUSION: Percutaneous intervention provides a promising alternative for patients with Type B aortic dissections when medical treatment will not improve the likelihood of meaningful recovery and surgery entails too great a risk. Nephrologists should therefore be aggressive in the workup of ischemic renal failure associated with aortic dissection as percutaneous intervention may reverse the effects of renal failure in this population. PMID- 15125783 TI - Antimicrobial activity of innate immune molecules against Streptococcus pneumoniae, Moraxella catarrhalis and nontypeable Haemophilus influenzae. AB - BACKGROUND: Despite its direct connection to the nasopharynx which harbors otitis media pathogens as part of its normal flora, the middle ear cavity is kept free of these bacteria by as yet unknown mechanisms. Respiratory mucosal epithelia, including those of the middle ear and eustachian tube, secrete antimicrobial effectors including lysozyme, lactoferrin and beta defensins-1 and -2. To elucidate the role of these innate immune molecules in the normal defense and maintenance of sterility of respiratory mucosa such as that of the middle ear, we assessed their effect on the respiratory pathogens nontypeable Haemophilus influenzae (NTHi) 12, Moraxella catarrhalis 035E, and Streptococcus pneumoniae 3, and 6B. METHODS: Two assay methods, the radial assay and the liquid broth assay, were employed for testing the antimicrobial activity of the molecules. This was done in order to minimize the possibility that the observed effects were artifacts of any single assay system employed. Also, transmission electron microscopy (TEM) was employed to evaluate the effect of antimicrobial innate immune molecules on OM pathogens. For the statistical analysis of the data, Student's t-test was performed. RESULTS: Results of the radial diffusion assay showed that beta defensin-2 was active against all four OM pathogens tested, while treatment with beta defensin-1 appeared to only affect M. catarrhalis. The radial assay results also showed that lysozyme was quite effective against S. pneumoniae 3 and 6B and was partially bacteriostatic/bactericidal against M. catarrhalis. Lysozyme however, appeared not to affect the growth of NTHi. Thus, lysozyme seems to have a more pronounced impact on the growth of the Gram positive S. pneumoniae as compared to that of Gram-negative pathogens. Lactoferrin on the other hand, enhanced the growth of the bacteria tested. The results of the radial assays were confirmed using liquid broth assays for antimicrobial activity, and showed that lysozyme and beta defensin-2 could act synergistically against S. pneumoniae 6B. Moreover, in the liquid broth assay, beta defensin-1 showed a modest inhibitory effect on the growth of S. pneumoniae 6B. As assessed by ultrastructural analysis, lysozyme and beta defensin-2, and to a much lesser extent, beta defensin-1, appeared to be able to cause damage to the bacterial membranes. CONCLUSIONS: Here we report that lysozyme and the beta defensins can inhibit the growth of clinical isolates of otitis media pathogens - namely NTHi strain 12, S. pneumoniae strains 3 and 6B and M. catarrhalis strain 035E - and cause ultrastructural damage to these pathogens. Moreover, we demonstrate that lysozyme and beta defensin-2 can act synergistically against S. pneumoniae. These findings are consistent with the concept that secreted antimicrobial peptides and other components of innate immunity constitute the first line of defense protecting host mucosal surfaces, including the tubotympanal (eustachian tube and middle ear cavity) mucosa, against pathogens. PMID- 15125784 TI - The influence of statins on the free intracellular calcium concentration in human umbilical vein endothelial cells. AB - BACKGROUND: Statins are cholesterol-lowering drugs that are widely used to reduce the risk of cardiac infarction. Their beneficial clinical effects, however, are not restricted to their influence on cholesterol production. As several studies have shown that they have a potency of relaxing blood vessels. METHODS: We measured the effects of statins on the intracellular free calcium concentration ([Ca2+]i) in human umbilical vein endothelial cells (HUVEC) after acute application and 24-h-preincubation of statins. RESULTS: Incubation of the cells for 24 h with cerivastatin or fluvastatin significantly increased the resting [Ca2+]i. For cerivastatin this effect manifested at a concentration of 1 microM. Increase of resting [Ca2+]i in the presence of cerivastatin also occurred when the nitric oxide synthase was inhibited. Transient Ca2+ release induced by histamine was not affected. CONCLUSIONS: The increase of resting [Ca2+]i after incubation with cerivastatin or fluvastatin may provide an explanation for the direct effects of statins on the endothelial-dependent vasodilatation and restoration of endothelial activity in vivo. PMID- 15125785 TI - Glucocorticoids synergize with IL-1beta to induce TLR2 expression via MAP Kinase Phosphatase-1-dependent dual Inhibition of MAPK JNK and p38 in epithelial cells. AB - BACKGROUND: Despite the importance of glucocorticoids in suppressing immune and inflammatory responses, their role in enhancing host immune and defense response against invading bacteria is poorly understood. Toll-like receptor 2 (TLR2) has recently gained importance as one of the major host defense receptors. The increased expression of TLR2 in response to bacteria-induced cytokines has been thought to be crucial for the accelerated immune response and resensitization of epithelial cells to invading pathogens. RESULTS: We show that IL-1beta, a key proinflammatory cytokine, greatly up-regulates TLR2 expression in human epithelial cells via a positive IKKbeta-IkappaBalpha-dependent NF-kappaB pathway and negative MEKK1-MKK4/7-JNK1/2 and MKK3/6-p38 alpha/beta pathways. Glucocorticoids synergistically enhance IL-1beta-induced TLR2 expression via specific up-regulation of the MAP kinase phosphatase-1 that, in turn, leads to dephosphorylation and inactivation of both MAPK JNK and p38, the negative regulators for TLR2 induction. CONCLUSION: These results indicate that glucocorticoids not only suppress immune and inflammatory response, but also enhance the expression of the host defense receptor, TLR2. Thus, our studies may bring new insights into the novel role of glucocorticoids in orchestrating and optimizing host immune and defense responses during bacterial infections and enhance our understanding of the signaling mechanisms underlying the glucocorticoid-mediated attenuation of MAPK. PMID- 15125786 TI - Female premenopausal fracture risk is associated with gc phenotype. AB - The phenotype of the vitamin D binding and macrophage activating protein, Gc, is a predictor of premenopausal bone fracture risk, possibly mediated through activation of osteoclasts. This was concluded from a study on 595 Danish perimenopausal women 45-58 years of age (30,040 person years). INTRODUCTION: The multifunctional plasma protein Gc, also known as group-specific component, Gc globulin, or vitamin D binding protein (DBP), has two functions with relation to bone tissue: it is the major carrier protein of vitamin D in the circulation, and deglycosylation converts it into a very potent macrophage- and osteoclast activating factor (Gc-MAF). There are several phenotypes of Gc, and in this study, we examined the relation between Gc phenotype and bone fragility. MATERIALS AND METHODS: By isoelectric focusing we identified the Gc phenotype of 595 white recent postmenopausal women enrolled into the Danish Osteoporosis Prevention Study (DOPS) and identified three groups: Gc1-1 (n = 323), Gc1-2 (n = 230), and Gc2-2 (n = 42). Differences between the three groups were examined with respect to number of fractures before enrollment, BMC and BMD, and various biochemical and clinical parameters, including the concentration of Gc measured by immunonephelometry and the concentration of the macrophage marker soluble CD163 measured by ELISA. RESULTS AND CONCLUSIONS: The risk of having at least one premenopausal bone fracture (total number of women with fracture = 179) differed significantly (p = 0.017) in women with phenotype Gc1-1 (110/323 = 0.34), Gc1-2 (63/230 = 0.27), and Gc2-2 (6/42 = 0.14). The differences were even more striking (p = 0.005) for fractures caused by low-energy traumas. Using logistic regression, we found the relative risk of premenopausal fracture to be 0.32 (0.13 0.80) in Gc2-2 compared with Gc1-1. We propose that the Gc phenotypes cause differences in osteoclast activity, a theory supported by our finding of lower levels of Gc and of soluble CD163 in women with Gc2-2 compared with Gc1-1. PMID- 15125787 TI - Genetically modified animal models as tools for studying bone and mineral metabolism. AB - Genetic modification of mice is a powerful tool for the study of bone development and metabolism. This review discusses the advantages and disadvantages of various approaches used in bone-related research and the contributions these studies have made to bone biology. Genetic modification of mice is a powerful tool for the study of bone development and metabolism. This review discusses the advantages and disadvantages of various approaches used in bone-related research and the contributions these studies have made to bone biology. The approaches to genetic modification included in this review are (1) overexpression of genes, (2) global gene knockouts, (3) tissue-specific gene deletion, and (4) gene knock-in models. This review also highlights issues that should be considered when using genetically modified animal models, including the rigorous control of genetic background, use of appropriate control lines, and confirmation of tissue specificity of gene expression where appropriate. This technology provides a unique and powerful way to probe the function of genes and is already revolutionizing our approach to understanding the physiology of bone development and metabolism. PMID- 15125788 TI - A meta-analysis of prior corticosteroid use and fracture risk. AB - The relationship between use of corticosteroids and fracture risk was estimated in a meta-analysis of data from seven cohort studies of approximately 42,000 men and women. Current and past use of corticosteroids was an important predictor of fracture risk that was independent of prior fracture and BMD. INTRODUCTION: The aims of this study were to validate that corticosteroid use is a significant risk factor for fracture in an international setting and to explore the effects of age and sex on this risk. MATERIALS AND METHODS: We studied 42,500 men and women from seven prospectively studied cohorts followed for 176,000 patient-years. The cohorts comprised the EPOS/EVOS study, CaMos, the Rotterdam Study, Dubbo Osteoporosis Epidemiology Study (DOES), and prospective cohorts at Sheffield, Rochester, and Gothenburg. The effect of ever use of corticosteroids, BMD, age, and sex on all fracture, osteoporotic fracture, and hip fracture risk alone was examined using Poisson regression in each cohort and for each sex. The results of the different studies were merged from the weighted beta coefficients. RESULTS: Previous corticosteroid use was associated with a significantly increased risk of any fracture, osteoporotic fracture, and hip fracture when adjusted for BMD. Relative risk of any fracture ranged from 1.98 at the age of 50 years to 1.66 at the age of 85 years. For osteoporotic fracture, the range of relative risk was 2.63-1.71, and for hip fracture 4.42-2.48. The estimate of relative risk was higher at younger ages, but not significantly so. No significant difference in risk was seen between men and women. The risk was marginally and not significantly upwardly adjusted when BMD was excluded from the model. The risk was independent of prior fracture. In the three cohorts that documented current corticosteroid use, BMD was significantly reduced at the femoral neck, but fracture risk was still only partly explained by BMD. CONCLUSION: We conclude that prior and current exposure to corticosteroids confers an increased risk of fracture that is of substantial importance beyond that explained by the measurement of BMD. Its identification on an international basis validates the use of this risk factor in case-finding strategies. PMID- 15125789 TI - Cadmium exposure and distal forearm fractures. AB - The aim of this study was to analyze the relationship between low-level cadmium exposure and distal forearm fractures. Altogether, 1021 men and women exposed to cadmium in Sweden were included. The study indicates that cadmium exposure is associated with increased risk of forearm fractures in people over the age of 50. INTRODUCTION: Very few studies have been performed on environmental risk factors for fractures. Cadmium is known to cause damage to the kidneys and in high doses to the bone. The aim of this study was to analyze the relationship between low level cadmium exposure and distal forearm fractures. MATERIALS AND METHODS: A total of 479 men and 542 women, 16-81 years of age, that were environmentally or occupationally exposed to cadmium were examined in 1997. Cadmium in urine was used to estimate dose, and information about previous fractures and risk factors for fractures was obtained from questionnaires. Fractures were validated using medical records. The association between cadmium dose and risk of forearm fracture was evaluated using Cox proportional hazard regression analysis. RESULTS AND CONCLUSION: The mean urinary cadmium in the study population was 0.74 nmol cadmium/mmol creatinine (10% and 90% percentiles are 0.19 and 1.42, respectively). For fractures occurring after the age of 50 years (n = 558, 32 forearm fractures), the fracture hazard ratio, adjusted for gender and other relevant co-variates, increased by 18% (95% CI, 1.0-38%) per unit urinary cadmium (nmol cadmium/mmol creatinine). When subjects were grouped in exposure categories, the hazard ratio reached 3.5 (90% CI, 1.1, 11) in the group of subjects with urinary cadmium between 2 and 4 nmol/mmol creatinine and 8.8 (90% CI, 2.6, 30) in the group of subjects with > or = 4 nmol/mmol creatinine. Associations between cadmium and fracture risk were absent before the age of 50. Cadmium exposure is associated with increased risk of forearm fractures in people over 50 years of age. PMID- 15125790 TI - Genetics of bone mineral density: evidence for a major pleiotropic effect from an intercontinental study. AB - BMD is a primary predictor of osteoporotic fracture, and its genetic determination is still unclear. This study showed that the correlation between BMD at different skeletal sites is caused by an underlying genetic structure of common genetic effects. In addition to possible shared (pleiotropic) genetic and environmental effects, each of the BMD variables may also be determined by site specific genetic factors. INTRODUCTION: BMD is a primary predictor of osteoporotic fracture and a key phenotype for the genetic study of osteoporosis. The interindividual variation in BMD measured at a given skeletal site is largely regulated by genetic factors. A strong phenotypic covariation exists for BMD at different skeletal sites. This study tests the hypothesis that the covariation is in fact caused by an underlying genetic structure of common genetic effects and that, in addition to possible shared (pleiotropic) genetic effects, each of the BMD variables may also be determined by site-specific genetic factors MATERIALS AND METHODS: A bivariate complex segregation analysis as implemented in statistical package PAP was conducted to explore various models of pleiotropic genetic and environmental transmission in lumbar spine and femoral neck BMD, as well as in compact and spongious segments of hand phalanges. The BMD was obtained in three ethnically, culturally, and socially heterogeneous samples of white pedigrees, with 2549 individuals between 18 and 100 years of age, from Australia, Europe, and North America. RESULTS AND CONCLUSIONS: The genetic correlation between BMD measures ranged between 0.50 +/- 0.09 and 0.79 +/- 0.04 in the three samples. In each sample, the model incorporated a major locus pleiotropic effect, and residual correlation was found to be the most parsimonious model. Estimated parameters from the model indicated a significant pleiotropic major gene effect on both lumbar spine and femoral neck BMD, with the existence of a significant residual correlation (0.51 +/- 0.07 to 0.66 +/- 0.04). These results suggest that the covariation in BMD at different skeletal sites, and between mostly compact versus mostly trabecular bone, was largely determined by common genetic factors that are pleiotropic or in close linkage and linkage disequilibirum, while at the same time, exhibiting considerable evidence of shared environmental effects. The results, for the first time, suggest that the possibility of pleiotropic genetic effect may be controlled by a major genetic locus. Identification of the major locus could open new opportunity to understanding the liability and pathogenic processes in which they are involved in the determination of fracture risk. PMID- 15125791 TI - Assessing bone status beyond BMD: evaluation of bone geometry and porosity by quantitative ultrasound of human finger phalanges. AB - In an in vitro study, we found significant associations between QUS variables and properties and geometrical parameters of the compact bone of human finger phalanges. QUS variables were not only related to BMD but also to other skeletal properties, which explained 70% of the variability of speed of sound. INTRODUCTION: Transverse transmission quantitative ultrasound (QUS) measurements at the finger phalanges are known to be correlated with BMD and to predict osteoporotic fractures. To determine which other skeletal properties are affected by ultrasound, we investigated the impact of density, geometry, and porosity on QUS variables in vitro. MATERIALS AND METHODS: Ultrasound variables were correlated with density, porosity, and geometrical characteristics of cortical bone. Additionally, we tested which combinations of geometry and bone properties best predicted the ultrasound results observed. Forty-four proximal phalanges from the middle finger were investigated at their distal metaphysis, similar to the typical clinical measurement procedure. Donor age ranged from 52 to 98 years (15 males and 29 females; mean age, 80.9 +/- 9.4 years). QUS variables were measured on the metaphysis of the phalanges using the DBMSonic 1200. Quantitative CT was used for the measurement of BMD, and high-resolution MRI was used for the measurement of porosity and geometrical variables. RESULTS: Speed of sound (SOS) and the clinically used variable AD-SOS correlated significantly with area, relative area, density, and porosity of the compact bone (R2 = 0.28-0.58, p < 0.0001). Signal amplitude correlated significantly only with relative area of the compact bone and area of the medullary canal (R2 = 0.18-0.20, p < 0.01). The combination of cortical area, density, and porosity improved the determination of SOS to R2 = 0.70, with a residual unexplained variability of 54 m/s (3.2%). CONCLUSIONS: These results clarify the impact of skeletal properties on QUS variables. SOS is affected by cortical area, cortical bone density, and cortical porosity, whereas attenuation only depends on geometry of the medulla. AD-SOS, the variable routinely measured in clinical practice, is primarily affected by cortical area. QUS of the finger phalanges is not only related to BMD but also to other skeletal properties. PMID- 15125792 TI - Maintenance of increased bone mass after recombinant human parathyroid hormone (1 84) with sequential zoledronate treatment in ovariectomized rats. AB - The concept of lose, restore, maintain (LRM) for reversing existing osteoporosis was tested in rats. The withdrawal of PTH results in the loss of the acquired bone mass, but sequential therapy with zoledronate quite effectively maintained the PTH(1-84)-acquired bone quantity and quality. INTRODUCTION: Because antiresorptive agents against osteoporosis are presently quite limited, strong anabolic agents such as human parathyroid hormone (hPTH) are quite helpful. However, because hPTH(1-34) is available only through injection and has a critical side effect of causing bone tumors during life-long administration in the rat, it would be practical to use PTH for the shortest possible duration to obtain the maximal effect. To determine the effectiveness of the osteoporosis reversing concept of lose, restore, and maintain (LRM), recombinant hPTH(1-84) [rhPTH(1-84)] and the respective antiresorptive agents were sequentially studied. MATERIALS AND METHODS: Thirty-six, 20-week-old Sprague-Dawley rats were used. Treatment started at the 25th week after ovariectomy, which was performed at 20 weeks of age, with 5 weeks of rhPTH(1-84) 100 microg/kg/day, 5 days/week, followed by the respective sequential therapies for 5 weeks as follows: (1) ovariectomized rats (OVX; n = 6), (2) sham-operated rats (SHAM; n = 6), (3) OVX rats with PTH maintenance (PTH-M; n = 6), (4) OVX rats treated with PTH and then PTH was withdrawn (PTH-W; n = 6), (5) PTH-treated OVX rats treated with 17beta estradiol (PTH-E; 10 microg/day SC, 5 days/week; n = 6), and (6) PTH-treated OVX rats treated with zoledronate (PTH-Z; 12.5 microg/kg SC weekly; n = 6). BMD of the right femora was measured by DXA. microCT was used to measure the structural parameters of the second lumbar vertebrae. Three-point bending test of the femora and compressive tests of vertebrae were also performed. RESULTS: Bone quantity data showed that the BMD and most of the microstructural parameters were significantly higher in the PTH-M and PTH-Z groups than in the OVX and PTH-W groups (p < 0.05). Measurement of the cortical thickness revealed that only the PTH-M group showed a significant increase (p = 0.001). The ultimate force (Fu) at the midshaft of the femora was similar in the treated groups and stronger than in the OVX group (p < 0.05). However, in the vertebrae, the Fu of the PTH-M and PTH Z groups was significantly higher, by approximately 44-47%, than in the OVX and PTH-E groups and showed a higher tendency than in the PTH-W group. CONCLUSION: PTH withdrawal resulted in the loss of acquired BMD, and sequential therapy with antiresorptives prevented further loss (17beta-estradiol versus zoledronate). The zoledronate after rhPTH(1-84) as a sequential regimen was quite consistently effective. PMID- 15125793 TI - Identification of novel regulators associated with early-phase osteoblast differentiation. AB - Key regulatory components of the BMP-induced osteoblast differentiation cascade remain to be established. Microarray and subsequent expression analyses in mice identified two transcription factors, Hey1 and Tcf7, with in vitro and in vivo expression characteristics very similar to Cbfa1. Transfection studies suggest that Tcf7 modulates BMP2-induced osteoblast differentiation. This study contributes to a better definition of the onset of BMP-induced osteoblast differentiation. INTRODUCTION: Elucidation of the genetic cascade guiding mesenchymal stem cells to become osteoblasts is of extreme importance for improving the treatment of bone-related diseases such as osteoporosis. The aim of this study was to identify regulators of the early phases of bone morphogenetic protein (BMP)2-induced osteoblast differentiation. MATERIALS AND METHODS: Osteoblast differentiation of mouse C2C12 cells was induced by treatment with BMP2, and regulation of gene expression was studied during the subsequent 24 h using high-density microarrays. The regulated genes were grouped by means of model-based clustering, and protein functions were assigned. Real-time quantitative RT-PCR analysis was used to validate BMP2-induced gene expression patterns in C2C12 cells. Osteoblast specificity was studied by comparing these expression patterns with those in C3H10T1/2 and NIH3T3 cells under similar conditions. In situ hybridization of mRNA in embryos at embryonic day (E)14.5 and E16.5 of gestation and on newborn mouse tails were used to study in vivo expression patterns. Cells constitutively expressing the regulated gene Tcf7 were used to investigate its influence on BMP-induced osteoblast differentiation. RESULTS AND CONCLUSIONS: A total of 184 genes and expressed sequence tags (ESTs) were differentially expressed in the first 24 h after BMP2 treatment and grouped in subsets of immediate early, intermediate early, and late early response genes. Signal transduction regulatory factors mainly represented the subset of immediate early genes. Regulation of expression of these genes was direct, independent of de novo protein synthesis and independent of the cell type studied. The intermediate early and late early genes consisted primarily of genes related to processes that modulate morphology, basement membrane formation, and synthesis of extracellular calcified matrix. The late early genes require de novo protein synthesis and show osteoblast specificity. In vivo and in vitro experiments showed that the transcription factors Hey1 and Tcf7 exhibited expression characteristics and cell type specificity very similar to those of the osteoblast specific transcription factor Cbfa1, and constitutive expression of Tcf7 in C2C12 cells differentially regulated osteoblast differentiation marker genes. PMID- 15125794 TI - Shockwave stimulates oxygen radical-mediated osteogenesis of the mesenchymal cells from human umbilical cord blood. AB - Human umbilical cord blood (HUCB) mesenchymal progenitor cells expressed stro-1 or CD44 or CD29, and subsequently, differentiated toward osteogenic lineage. Physical shockwave treatment increased osteogenic activity of HUCB mesenchymal progenitor cells through superoxide-mediated TGF-beta1 induction. Transplantation of shockwave-treated HUCB mesenchymal progenitor cells enhanced healing of segmental femoral defect in severe combined immunodeficiency disease (SCID) mice. INTRODUCTION: Mesenchymal progenitor cells (MPCs) in the bone marrow are precursors to bone development. It remains uncertain whether MPCs are present in human umbilical cord blood (HUCB) and are capable of differentiating into osteogenic cell lineage. Extending from a model of shockwave (SW) promotion of bone marrow stromal cell differentiation toward osteoprogenitors in rats, we further investigated how physical SW mediated biological responses in regulating osteogenic differentiation of HUCB MPCs. MATERIALS AND METHODS: HUCB was subjected to SW treatment at different energy flux densities and impulses. Colony forming units-stroma (CFU-Stroma), osteogenic activities (Cbfa1/Runx2 expression, bone alkaline phosphatase activity, and bone nodule formation), and bone formation by heterologous transplantation into SCID mice were assessed. RESULTS: Few CD34+ stem cells (1.3%) and stro-1+ cells (1.0%) were present in the freshly prepared mononuclear cells (MNCs) from HUCB. The number of stro-1+ cells, but not CD34+, increased to 72.4% in the adherent cell culture over 6 days. Stro-1+ cells co-expressed CD44 and CD29 markers and grew into CFU-Stroma that matured into bone nodules. We found that the SW treatment (0.16 mJ/mm2 energy flux density, 200 impulses) elicited superoxide production and promoted formation of CFU Stroma, but not of hematopoietic CFU-Mix. SW also enhanced the production of transforming growth factor (TGF)-beta1, but not of interleukin (IL)-3 or granulocyte monocyte-colony stimulating factor (GM-CSF). Neutralization of TGF beta1 significantly reduced SW-promoted CFU-Stroma formation. Superoxide scavenging by superoxide dismutase blocked SW enhancement of TGF-beta1 production and formation of CFU-Stroma. Administration of SW-treated HUCB MPCs to SCID mice with femoral segmental defects facilitated dense, bridging callus and gap closure. CONCLUSION: HUCB MPCs subjected to SW treatment is a potential source for stem cells useful in the treatment of orthopedic disorders. An optimal physical SW treatment enhanced osteogenesis through superoxide-mediated TGF-beta1 production. Physical stimulation is an alternative method for extending mesenchymal stem cells of HUCB. PMID- 15125795 TI - NF1 tumor suppressor protein and mRNA in skeletal tissues of developing and adult normal mouse and NF1-deficient embryos. AB - NF1 is a heritable disease with multiple osseous lesions. The expression of the NF1 gene was studied in embryonic and adult rodent skeleton and in NF1-deficient embryos. The NF1 gene was expressed intensely in the cartilage and the periosteum. Impaired NF1 expression may lead to inappropriate development and dynamics of bones and ultimately to the osseous manifestations of the disease. INTRODUCTION: Neurofibromatosis type 1 is caused by mutations in the NF1 gene encoding the Ras GTPase activating protein (Ras-GAP) neurofibromin. Skeletal ailments such as short stature, kyphoscoliosis, and tibial bowing and pseudarthrosis are common osseous manifestations of NF1. These symptoms are congenital, implying a role for neurofibromin in proper bone growth. However, little is known about its expression in skeletal tissues during their development. MATERIALS AND METHODS: The expression of the NF1 gene was studied in normal and NF1+/- mouse fetuses at embryonic days 12.5-15.5 and in skeletal tissues of adult mice and rats. In situ hybridization, immunohistochemistry, and Western blot analysis were used to identify the NF1 gene expression profile. RESULTS: NF1 mRNA and protein were elevated in resting, maturation, and hypertrophic chondrocytes at the growth plate. Parallel studies on NF1+/- embryos showed expression patterns identical to wildtype. The periosteum, including osteoblasts and osteoclasts, and osteocytes of the cortical bone of adult mice were also intensely labeled for NF1 protein and mRNA. Western transfer analysis detected NF1 protein in the respective rat tissues. Phosphorylation of p42 and p44 MAP kinases, the downstream consequence of Ras activation, was elevated in hypertrophic chondrocytes of NF1+/- embryos. CONCLUSIONS: The results suggest that neurofibromin may act as a Ras-GAP in skeletal cells to attenuate Ras transduced growth signals and thus play a role during ossification and dynamics of bone. Loss of NF1 function may therefore lead to dysplastic bone growth, thereby causing the debilitating osseous symptoms of NF1. PMID- 15125796 TI - Arthritis induces lymphocytic bone marrow inflammation and endosteal bone formation. AB - Arthritis can destroy the cortical bone barrier and expose bone marrow to synovial tissue. This study examines bone marrow changes in arthritis and its effects on cortical bone remodeling. Bone marrow next to arthritic lesions exhibits B-lymphocyte-rich infiltrates, which express BMPs and stimulate endosteal bone formation. Thus, bone marrow actively participates in the arthritic process. INTRODUCTION: Imaging studies have shown that bone marrow changes occur in patients with rheumatoid arthritis (RA). To examine whether bone marrow is affected during arthritis, human TNF transgenic (hTNFtg) mice, which constitute an established animal model of human RA, were examined for bone marrow changes. MATERIALS AND METHODS: The hind paws (tarsal area) of 22 untreated hTNFtg mice, 5 hTNFtg mice treated with anti-TNF (infliximab), and 5 wildtype (WT) mice were examined histologically, immunohistochemically, and by means of mRNA in situ hybridization. RESULTS AND CONCLUSIONS: All untreated hTNFtg mice with moderate (n = 10) and severe (n = 7) disease developed inflammatory bone marrow lesions during the course of disease, whereas no such lesions appeared in hTNFtg mice with mild disease (n = 5) and WT mice. Bone marrow infiltrates were almost exclusively composed of lymphocytes, and the overwhelming proportion (>80%) was B-cells. Presence and extent of bone marrow infiltrates were closely linked to severity of arthritis. In addition, blockade of TNF effectively reduced bone marrow inflammation. Interestingly, osteoblast numbers were increased at the endosteal surface in the vicinity of these lesions. Moreover, osteoid deposition; expression of bone matrix proteins, such as osteocalcin and osteopontin; and mineralization were enhanced, suggesting that inflammatory bone marrow infiltrates induce bone formation. Indeed, B-lymphocytes of these lesions expressed bone morphogenetic protein (BMP)-6 and -7, which are important stimulators of new bone formation. Thus, we conclude that bone marrow actively participates in destructive arthritis by generating B-lymphocyte-rich bone marrow lesions and inducing endosteal bone formation. PMID- 15125797 TI - Suppressed bone turnover by long-term bisphosphonate treatment accumulates microdamage but maintains intrinsic material properties in cortical bone of dog rib. AB - Effects of long-term suppression of bone remodeling by bisphosphonate were investigated in cortical bone of dog rib. Although microdamage was accumulated, BMD was increased without increasing cortical bone area. Consequently, the intrinsic material properties were not reduced. INTRODUCTION: Recently, we have reported that long-term suppression of bone remodeling increases microdamage accumulation but is not necessarily associated with vertebral fragility because of compensated increase of bone mass and improved microarchitecture. This study aimed to investigate the effect of long-term suppression of bone remodeling by bisphosphonate on the degree of mineralization, accumulation of microdamage, and mechanical properties of cortical bone in the same dogs. MATERIALS AND METHODS: Twenty-nine 1-year-old beagles (15 males, 14 females) were divided into three groups and treated daily with vehicle (CNT) or with incadronate at a dose of 0.3 (LOW) or 0.6 mg/kg/day (HIGH) orally for 3 years. After death, pQCT, histomorphometry, microdamage measurements, and three-point bending mechanical test were performed using the ninth rib. RESULTS: Cortical BMD was increased in the incadronate-treated groups. Cortical activation frequency was suppressed by 82% and 70% in HIGH and LOW, respectively, compared with CNT, without impairment of mineralization. Microdamage accumulation was increased in both incadronate treated groups. Although there were no significant differences in total and cortical area among the three groups, structural mechanical properties were significantly increased after incadronate treatment while intrinsic material properties were not changed in the incadronate-treated groups. CONCLUSION: This study suggests that long-term suppression of bone remodeling by bisphosphonate increases microdamage accumulation. However, this was not necessarily associated with a reduction of intrinsic material properties probably because of an increased degree of mineralization. PMID- 15125798 TI - Cortical and trabecular bone mineral loss from the spine and hip in long-duration spaceflight. AB - We measured cortical and trabecular bone loss using QCT of the spine and hip in 14 crewmembers making 4- to 6-month flights on the International Space Station. There was no compartment-specific loss of bone in the spine. Cortical bone mineral loss in the hip occurred primarily by endocortical thinning. INTRODUCTION: In an earlier study, areal BMD (aBMD) measurements by DXA showed that cosmonauts making flights of 4- to 12-month duration on the Soviet/Russian MIR spacecraft lost bone at an average rate of 1%/month from the spine and 1.5%/month from the hip. However, because DXA measurements represent the sum of the cortical and trabecular compartments, there is no direct information on how these bone envelopes are affected by spaceflight. MATERIALS AND METHODS: To address this, we performed a study of crewmembers (13 males and 1 female; age range, 40-55 years) on long-duration missions (4-6 months) on the International Space Station (ISS). We used DXA to obtain aBMD of the hip and spine and volumetric QCT (vQCT) to assess integral, cortical, and trabecular volumetric BMD (vBMD) in the hip and spine. In the heel, DXA was used to measure aBMD, and quantitative ultrasound (QUS) was used to measure speed of sound (SOS) and broadband ultrasound attenuation (BUA). RESULTS AND CONCLUSIONS: aBMD was lost at rates of 0.9%/month at the spine (p < 0.001) and 1.4-1.5%/month at the hip (p < 0.001). Spinal integral vBMD was lost at a rate of 0.9%/month (p < 0.001), and trabecular vBMD was lost at 0.7%/month (p < 0.05). In contrast to earlier reports, these changes were generalized across the vertebrae and not focused in the posterior elements. In the hip, integral, cortical, and trabecular vBMD was lost at rates of 1.2-1.5%/month (p < 0.0001), 0.4-0.5%/month (p < 0.01), and 2.2 2.7%/month (p < 0.001), respectively. The cortical bone loss in the hip occurred primarily by cortical thinning. Calcaneal aBMD measurements by DXA showed smaller mean losses (0.4%/month) than hip or spine measurements, with SOS and BUA showing no change. In summary, our results show that ISS crewmembers, on average, experience substantial loss of both trabecular and cortical bone in the hip and somewhat smaller losses in the spine. These results do not support the use of calcaneal aBMD or QUS measurements as surrogate measures to estimate changes in the central skeleton. PMID- 15125800 TI - Does work stress predict insomnia? A prospective study. AB - OBJECTIVES: The aim of this study was to investigate the one-year development of self-reported sleep problems in workers with no sleep problem at baseline, and to evaluate the role of work stress in the etiology of a new episode. DESIGN: A prospective design was employed. METHODS: A total of 816 employees with no sleeping problems during the past three months completed a baseline questionnaire concerning their general health, working hours and working conditions. One year later they were contacted again to ascertain whether they were experiencing problems sleeping. RESULTS: At the follow-up, the three-month point prevalence of self-reported sleep problems for this population was 14.3%. While controlling for age and gender, it was found that irregular working hours and general health were not significantly related to the development of a new episode of sleeping problems. However, stress in the form of a 'poor' psychosocial work environment increased the risk of a new episode by more than twofold (odds ratio 2:15). The attributable fraction suggested that eliminating stress could prevent 53% of the cases. CONCLUSION: In a population of employees with no reported sleeping problems, 14.3% developed a sleeping problem during the coming year. Even when controlling for possible confounders, stress in the form of a 'poor' psychosocial work environment doubled the risk of developing a sleep problem. PMID- 15125799 TI - Two novel mutations at exon 8 of the Sequestosome 1 (SQSTM1) gene in an Italian series of patients affected by Paget's disease of bone (PDB). AB - PDB is genetically heterogeneous. Mutations of the sequestosome1 gene have been reported in sporadic and familial forms of Paget's in patients of French Canadian and British descent. Mutational analyses in different ethnic groups are needed to accurately investigate hereditary diseases. We describe two novel mutations of sequestosome1 in 62 Italian sporadic patients, confirming the role of the encoded protein in this disorder. INTRODUCTION: Paget's disease of bone (PDB) is a relatively common disease of bone metabolism reported to affect up to 3% of whites over 55 years of age. The disorder is genetically heterogeneous, and at present, there is scientific evidence that at least eight different human chromosomal loci are correlated with its pathogenesis. Mutations of the sequestosome1 (SQSTM1) gene were identified as responsible for most of the sporadic and familial forms of Paget in patients of French Canadian and British descent. Such mutations were located at exon 7 and 8 levels, encoding for the ubiquitin protein-binding domain (UBA) and representing a mutational hot spot area. MATERIALS AND METHODS: To verify the involvement of this gene in Italian subjects affected by PDB, we performed mutational analysis in 62 sporadic PDB cases. RESULTS: We described three different mutations at exon 8 level: P392L, already described in the French Canadian population and families predominantly of British descendent, and two novel mutations consisting of the amino acid substitutions M404V and G425R. No significant differences in the clinical history of PDB have been observed in patients with SQSTM1 mutations in respect to those without. CONCLUSIONS: Even though our findings suggest a minor involvement of the SQSTM1 gene in the pathogenesis of sporadic Italian Paget's cases, the identification of different significant mutations within the SQSTM1 gene in unrelated, but clinically similar individuals, offers extremely convincing evidence for a causal relationship between this gene and PDB. Longitudinal studies are needed to assess the penetrance of genotype/phenotype correlations. Our findings confirm the evidence of a clustered mutation area at this level in this disorder. PMID- 15125801 TI - The role of hardiness in stress and illness: An exploration of the effect of negative affectivity and gender. AB - Past research into the effects of hardiness on stress and health has yielded inconsistent findings, possibly because of a failure to acknowledge the influence of variables such as negative affectivity and gender. This study examined the main, moderating and mediating effects of hardiness in a sample of 130 (50 male, 80 female) randomly selected university staff members. Controlling for negative affectivity, limited evidence was obtained for the direct effects of hardiness on stress and illness. Kobasa's (1979) model that hardiness buffers the effects of stress on illness was supported for males, but not for females. The hypothesis that approach and avoidance coping mediate the hardiness-illness relationship was not supported for either sex, although there was evidence that relative coping mediated this relationship in females. Collectively, the results point to a need to reconsider the conceptualization and measurement of hardiness. PMID- 15125802 TI - Positivity in illness: self-delusion or existential growth? AB - OBJECTIVES: This study investigated the relationship between a measure of positivity in illness, the Silver Lining Questionnaire (SLQ), and measures of personality and spirituality/religious beliefs as a way of determining whether positivity in illness is a delusion or existential growth. METHOD: This is a cross-sectional study comparing response to the SLQ, to the Eysenck Personality Questionnaire (EPQ-R), breathlessness, illness type, and spiritual and religious beliefs in a final total sample of 194 respiratory outpatients. RESULTS: The SLQ was associated positively with extraversion (r =.16, p<.05), unrelated to neuroticism (r =.11, n.s.) and repression (r =.10, n.s.) and was positively associated with spiritual and religious beliefs, F(2; 187) = 7.12, p < 001, as predicted by the existential growth but not the delusion interpretation. There was no relationship between positivity and age, r(194) =.09, n.s., or between positivity and gender t(192) = -1.27, n.s., and nor were there relationships with type of illness, F(4, 188) = 2.17, n.s., or breathlessness, F (5, 173) = 0.42, n.s. CONCLUSIONS: The results suggest that positivity in illness is associated with existential growth, though the cross-sectional nature of the study precludes a conclusion of causal direction. The non-significant correlation between the SLQ and neuroticism is in the opposite direction predicted by the delusion explanation, but the non-significant relationship between the SLQ and repression is in the predicted direction. We cannot rule out the possibility that some positivity is delusion. PMID- 15125803 TI - Dieting and working memory: preoccupying cognitions and the role of the articulatory control process. AB - OBJECTIVES: The study aimed to extend previous research that has demonstrated impaired cognitive function among dieters, by teasing out precisely which components of the phonological loop subsystem of working memory are affected by dieting. METHOD: Participants were 92 female undergraduate students divided into three groups; current dieters (n = 19), past dieters (n = 29) and never dieters (n = 44). They completed cognitive performance tasks designed to preferentially load on the phonological store and articulatory control process components of the phonological loop, as well as self-report measures. RESULTS: Current dieting to lose weight had no effect on the phonological store, as evidenced by the lack of effect on the phonological similarity effect. Dieting did, however, impede the articulatory control process, as evidenced by substantial reduction in the world length effect. This effect was partially mediated by preoccupying cognitions concerning food, weight and body shape. Further, the results mirrored those found with non-dieters under conditions of articulatory suppression. CONCLUSIONS: The results clearly identify the articulatory control process as the site of the cognitive impairment suffered by dieters, and thereby offer very strong support to the preoccupying cognitions concerning food, weight and body shape that accompany dieting behaviour as the underlying mechanism. PMID- 15125804 TI - The impact of personal and social comparison information about health risk. AB - OBJECTIVES: To examine the emotional and cognitive impact of personal and social comparison information about health risk. METHODS: A total of 970 adults responded to vignettes describing risk presentation scenarios that varied in terms of having (a) a 'real world' analogue (cardiac event) versus no such analogue (a fictitious pancreatic disease) condition, (b) high versus low levels of personal risk, and (c) no comparison group information given, comparison group risk higher or lower than own risk. RESULTS: For both the cardiac and pancreatic disease vignettes, respondents' emotional responses and estimates of their own risk were influenced by both personal and social comparison risk information. The cardiac event vignettes produced larger effects than the pancreatic disease vignettes. Unfavourable social comparison information had no discernible impact, relative to not providing any social comparison information. Favourable social comparison information resulted in greater reassurance, less worry, and perceptions of lower susceptibility. Lower personal risk generally produced similar effects, relative to higher personal risk. CONCLUSIONS: In contrast to previous theory and research in this area, we found that both personal and (favourable) social comparison risk information have emotional and cognitive consequences. We hypothesize that the perceived clarity of the information may account for the different patterns of findings in the literature. PMID- 15125805 TI - Health beliefs and communication in the travel clinic consultation as predictors of adherence to malaria chemoprophylaxis. AB - OBJECTIVES: The objectives were, first, to determine whether adherence to malaria prophylaxis could be predicted by (i) health beliefs specified by the Health Belief Model and the Theory of Planned Behaviour, and (ii) communication during the consultation in a travel clinic; and secondly, to examine the impact of the consultation in changing travellers' health beliefs. DESIGN: A prospective study using regression analysis. METHODS: The participants were 130 consecutive travellers attending a travel medicine clinic. Health beliefs were measured pre- and post-consultation. The consultations were coded from audiotape using the Roter Interaction Analysis System and a content analysis method recording discussion about malaria and prophylaxis. Adherence was assessed by a follow-up telephone interview. RESULTS: Perceived susceptibility to malaria, perceived benefits of medication and intentions to adhere increased significantly as a result of the consultation, and the perceived permanent nature of side effects reduced significantly. At follow-up (N = 107), 62% reported full adherence, 25% partial adherence and 12% poor/no adherence. A multinomial logistic regression analysis revealed that perceived benefits of medication, length of stay, health professional discussion about adherence and travellers' questions and statements independently predicted reported adherence. CONCLUSIONS: Health beliefs and communication significantly predicted adherence in this setting. The findings also suggested qualitative differences between travellers who adhered fully, partially or poorly. Although the clinic consultation had a positive impact, emphasizing benefits of medication and resolving potential barriers to adherence could improve adherence in the population. PMID- 15125806 TI - Individual differences in the temporal variability of medically unexplained symptom reporting. AB - THEORY: Non-specific or medically unexplained symptoms account for up to 35% of outpatient referrals. In contrast to the literature on affect, little is known about how people frame daily symptoms and how these patterns are influenced by individual differences. METHOD: Three fixed occasion diary studies over an 8-day period (one on ill veterans of the Gulf War and two on healthy undergraduates) and a single cross-sectional survey (on ill Gulf War veterans) were conducted. The severity and frequency of daily symptoms were assessed in all daily diary samples, together with the Big 5 personality domains in one of the undergraduate samples and in the survey of veterans. The data were analysed using auto correlations and hierarchical multivariate linear modelling. RESULTS: In all the chronically ill and healthy samples, the data suggested potential cyclical patterns for symptom severity reporting. With regard to the frequency of symptom reporting, the veterans showed a pattern of constant symptom reporting. Finally, the relationship between the reported severity of symptoms was attenuated by intellect and surgency. DISCUSSION: There is evidence that daily experiences of symptom severity are framed relative to each other and this relationship is influenced by personality. The practical implications of these findings are discussed. PMID- 15125807 TI - Using the IPQ and PMDI to predict regular diabetes care-seeking among patients with Type 1 diabetes. AB - OBJECTIVES: The objectives were, first, to identify factors associated with regular diabetes care-seeking and, second, to compare the performance of the Illness Perception Questionnaire (IPQ) and a modified version of the Personal Models of Diabetes Interview (PMDI) in predicting care-seeking. METHOD: This was a cross-sectional study involving 42 patients who had not attended hospital diabetes clinic for a period of 18 months or more and 42 matched controls receiving specialist care. Differences in illness representations between clinic attenders and non-attenders were examined. Due to the variability in care-seeking between non-attenders (ranging from no contact with health professionals to regular general practitioners care), participants were then reclassified for further analyses into those receiving regular care from either hospital diabetes clinic or general practice (n = 52) and those receiving no regular care (n = 32). RESULTS: Patients not seeking regular care held more negative views of the control, course and consequences of diabetes than those who received regular care. Regression analyses showed that the most important construct was treatment effectiveness. Treatment effectiveness (PMDI) and control (IPQ) dimensions were associated with clinic attendance. PMDI constructs of treatment effectiveness, threat and worries were associated with regular care-seeking. CONCLUSIONS: IPQ and PMDI results were generally consistent. In order to understand diabetes care seeking behaviour it is important to measure beliefs about the benefits of treatment recommendations in addition to those of the disease itself. Emotional and cognitive responses to diabetes should be assessed. PMID- 15125808 TI - Stages of motivational readiness for physical activity: a comparison of different algorithms of classification. AB - OBJECTIVE: The aim of this study was to compare two approaches to classify individuals into stages of motivational readiness for physical activity and test which one was better explained by attitude and perceived behavioural control, as defined by Ajzen (1991). DESIGN: A survey of 20,430 respondents from a population based sample. METHODS: The relevant variables were assessed in a self administered questionnaire. The cluster approach consisted of combining both intention and behaviour in order to determine clusters of individuals; such clusters correspond to different stages of motivational readiness. The stage of change (SC) approach consisted of grouping the same individuals by using the SC variable of the Transtheoretical Model (TTM). RESULTS: The SC and cluster solution approaches were replicated across four subsamples of the total number of respondents. Attitude and perceived behavioural control were more strongly associated with stage membership derived from four-cluster solution than with stage membership in the five categories assessed by the SC method. CONCLUSION: Stage of motivational readiness for physical activity, and possibly for other health-related behaviours, may usefully be characterized when both recent past behaviour and intention in the near future are simultaneously and explicitly taken into consideration. PMID- 15125810 TI - Delivering the diagnosis. PMID- 15125809 TI - Deciding to exercise: the role of anticipated regret. AB - OBJECTIVES: Two studies tested (a) whether anticipated regret (AR) qualifies as an additional predictor of intentions to engage in a health-enhancing behaviour (exercise) after variables from the Theory of Planned Behaviour (TPB) and past behaviour have been controlled, and (b) whether a manipulation that induces participants to focus on AR causes stronger intentions to exercise. DESIGN: Study 1 employed a cross-sectional questionnaire design; Study 2 employed a between participants experimental design (AR focus vs. no AR focus). METHOD: Participants (N = 385) completed standard, multi-item, reliable measures of TPB constructs and AR and also reported their past behaviour (Study 1). Participants in Study 2 (N = 70) completed measures of AR and intention; salience of AR was manipulated by means of item order. RESULTS: Study 1 showed that even though TPB variables and past behaviour were reliable predictors of intention, and explained 51% of the variance, AR contributed a substantial increment in the variance (5%) even after these predictors had been taken into account. Study 2 showed that participants who were induced to focus on AR prior to intention formation had significantly stronger intentions to exercise compared to controls. CONCLUSION: The findings indicate that AR predicts a health-enhancing behaviour (as well as the health risk behaviours examined in previous research) and that effects of AR are independent of TPB variables and past behaviour. The findings also indicate that a simple and inexpensive manipulation of the salience of AR can be used to promote exercise intentions. PMID- 15125811 TI - Paraclinical support of the person diagnosed with multiple sclerosis. AB - Receiving a diagnosis of multiple sclerosis (MS) is a life-altering event which has a profound impact on the patient. The author (a medical social worker for 13 years in a large MS clinic) suggests a model of paraclinical support for the person with MS and their family and friends, during and post-diagnosis. The model combines psychosocial assessment, information dissemination, emotional support and tangible interventions. The resultant patient-centred, holistic, longitudinal care aids adjustment to the diagnosis and pursuit of quality of life, despite the disease. PMID- 15125812 TI - Combination therapy for multiple sclerosis. AB - To improve the partial benefit of approved monotherapies in multiple sclerosis (MS), over 25 combined therapies have been tested in recent years, either as fixed-dose or sequential combination regimes. The main therapeutic targets for combination therapy in MS are the same as for monotherapy: the immune-mediated inflammatory cascade, oxidative toxicity and excitotoxicity. There are numerous reasons to consider combination therapy in MS, including to improve the benefit and/or tolerance in patients responding to approved treatments, stop frequent disabling relapses and/or rapid progression in patients who do not respond to approved therapies, and maintain the benefit of immunosuppressive treatments. Preliminary clinical trials suggest that combination therapy in MS does not increase the side-effects of approved monotherapy; its efficacy over monotherapy should therefore be tested. Statistically robust trials would need to involve many patients for each combination, so the first step in determining the efficacy of combination therapy should be to perform safety studies, followed by proof-of concept efficacy studies. PMID- 15125813 TI - Dysphagia and multiple sclerosis. AB - Over 30% of persons with multiple sclerosis (pwMS) suffer from swallowing symptoms, a higher rate than previously assumed. Neurogenic dysphagia (ND) may cause many different kinds of oropharyngeal sensorimotor dysfunctions in pwMS, and is associated with both the amount of disability and brainstem signs. About 15% of pwMS with mild disability may also suffer from ND. Diagnostic tools comprise history taking, bedside screening examination (50 ml water test combined with assessment of pharyngeal sensation or with pulse oximetry) and sometimes a videofluoroscopic swallowing study (VFSS) and fibreoptic endoscopic evaluation of swallowing (FEES). VFSS and FEES are complementary methods and both have advantages and disadvantages. Interventions for ND in pwMS are mainly based on functional swallowing therapy, including methods of restitution, compensation and adaptation. The aim of intervention is to prevent aspiration and aspiration pneumonia. Outcome assessment should focus on clinically relevant parameters, such as activity limitation, participation restriction and health-related quality of life. PMID- 15125814 TI - First Regional MS Forum Meeting raises MS awareness and highlights questions. The MS Forum Pan-Asian Conference, 912 October 2003, Bangkok, Thailand. AB - Multiple sclerosis (MS) in Asia and the Middle East was the subject of extensive discussion and debate at the first Pan-Asian MS Forum conference, entitled MS Across Continents: Insights from Asia and the Middle East. The first regional MS Forum activity, and the first major gathering of MS experts from these regions for over 30 years, it attracted over 100 specialists to share their experiences on the features, characteristics and management of the disease. PMID- 15125815 TI - Improving diabetes care. PMID- 15125820 TI - Chronic papules on the back and extremities. PMID- 15125821 TI - Strategies to reduce complications of type 2 diabetes. PMID- 15125823 TI - Treating type 2 diabetes: targeting the many causative factors. PMID- 15125824 TI - What the new Medicare prescription drug bill may mean for providers and patients. PMID- 15125825 TI - Starting insulin in type 2 diabetes: continue oral hypoglycemic agents? A randomized trial in primary care. AB - OBJECTIVE: To evaluate the effects of insulin 30/70 twice daily or bedtime isophane (NPH) insulin plus continued sulfonylurea and metformin in patients with type 2 diabetes in primary care. STUDY DESIGN: Open-label, randomized trial. POPULATION: Persons younger than 76 years with type 2 diabetes whose disease had not been controlled with oral hypoglycemic agents alone. A total of 64 insulin naive patients treated with maximal feasible dosages of sulfonylurea and metformin (baseline glycosylated hemoglobin [HbA1c]=8.5%) were randomly assigned to insulin monotherapy (IM group; n=31) or insulin in addition to unchanged oral hypoglycemic medication (IC group; n=33) for 12 months. Insulin doses were adjusted to obtain fasting glucose <7.0 mmol/L and postprandial glucose <10.0 mmol/L. OUTCOMES MEASURED: Outcome measures included HbA1c, treatment failure, weight, hypoglycemic events and symptoms, satisfaction with treatment, general well-being, and fear of injecting insulin and testing. RESULTS: HbA1c improved from 8.3% to 7.6% in the IC group, and from 8.8% to 7.6% in the IM group (P=NS). The IC group had 24% treatment failures, compared with 2% in the IM group (P=.09). Patients in the IC group had less weight gain than those in the IM group (1.3 vs 4.2 kg; P=.01), and they reported fewer hypoglycemic events (2.7 vs 4.3; P=.02). Increased satisfaction with treatment was equal in the 2 groups, and general well-being improved by 3.0 points more in the IC group (P=.05). Fear of self-injecting and self-testing did not differ. CONCLUSIONS: Bedtime NPH insulin added to maximal therapy with sulfonylurea and metformin is an effective, simple, well-tolerated approach for patients with uncontrolled type 2 diabetes. PMID- 15125826 TI - Clinical inquiries. Does screening for diabetes in at-risk patients improve long term outcomes? PMID- 15125827 TI - Clinical inquiries. What is the best treatment for diabetic neuropathy? PMID- 15125828 TI - Clinical inquiries. What is the best way to treat patients with white-coat hypertension? PMID- 15125829 TI - Clinical inquiries. Does a short symptom checklist accurately diagnose ADHD? PMID- 15125830 TI - Clinical inquiries. Should home apnea monitoring be recommended to prevent SIDS? PMID- 15125831 TI - Meralgia paresthetica is not always an entrapment neuropathy. PMID- 15125832 TI - Clearance of apoptotic cells: getting rid of the corpses. AB - The efficient elimination of apoptotic cells is crucial for tissue homeostasis in multicellular organisms. Secreted "find-me," exposed "eat-me," and lacking "don't eat-me" signals comprise the central elements of apoptotic cell removal, thus preventing the release of intracellular contents into the surrounding tissue. This is of special importance, as there is growing evidence that the onset of autoimmune disorders can be linked to the inefficient removal of apoptotic cells. This review focuses on the signals displayed by apoptotic cells, the bridging and receptor molecules on the phagocyte, and is intended to present a simplified model of the phagocytic synapse. Additionally, the recent discovery of lysophosphatidylcholine functioning as soluble attraction signal is discussed in the general context of apoptotic cell clearance. PMID- 15125833 TI - The TRAF6 ubiquitin ligase and TAK1 kinase mediate IKK activation by BCL10 and MALT1 in T lymphocytes. AB - The CARD domain protein BCL10 and paracaspase MALT1 are essential for the activation of IkappaB kinase (IKK) and NF-kappaB in response to T cell receptor (TCR) stimulation. Here we present evidence that TRAF6 ubiquitin ligase and TAK1 protein kinase mediate IKK activation by BCL10 and MALT1. RNAi-mediated silencing of MALT1, TAK1, TRAF6, and TRAF2 suppressed TCR-dependent IKK activation and interleukin-2 production in T cells. Furthermore, we have reconstituted the pathway from BCL10 to IKK activation in vitro with purified proteins of MALT1, TRAF6, TAK1, and ubiquitination enzymes including Ubc13/Uev1A. We find that a small fraction of BCL10 and MALT1 proteins form high molecular weight oligomers. Strikingly, only these oligomeric forms of BCL10 and MALT1 can activate IKK in vitro. The MALT1 oligomers bind to TRAF6, induce TRAF6 oligomerization, and activate the ligase activity of TRAF6 to polyubiquitinate NEMO. These results reveal an oligomerization --> ubiquitination --> phosphorylation cascade that culminates in NF-kappaB activation in T lymphocytes. PMID- 15125834 TI - Identification of beta-arrestin2 as a G protein-coupled receptor-stimulated regulator of NF-kappaB pathways. AB - Norepinephrine released by the sympathetic nerve terminals regulates the immune system primarily via its stimulation of beta(2)-adrenergic receptor (beta(2)AR), but the underlying molecular mechanisms remain to be elicited. Beta(2)AR, a well studied G protein-coupled receptor (GPCR), is functionally regulated by beta arrestin2, which not only causes receptor desensitization and internalization but also serves as a signaling molecule in GPCR signal transduction. Here we show that beta-arrestin2 directly interacts with IkappaBalpha (inhibitor of NF-kappaB, the key molecule in innate and adaptive immunity) and thus prevents the phosphorylation and degradation of IkappaBalpha. Consequently, beta-arrestin2 effectively modulates activation of NF-kappaB and expression of NF-kappaB target genes. Moreover, stimulation of beta(2)AR significantly enhances beta-arrestin2 IkappaBalpha interaction and greatly promotes beta-arrestin2 stabilization of IkappaBalpha, indicating that beta-arrestin2 mediates a crosstalk between beta(2)AR and NF-kappaB signaling pathways. Taken together, the current study may present a novel mechanism for regulation of the immune system by the sympathetic nervous system. PMID- 15125835 TI - Mitotic phosphorylation of the peripheral Golgi protein Nir2 by Cdk1 provides a docking mechanism for Plk1 and affects cytokinesis completion. AB - The rearrangement of the Golgi apparatus during mitosis is regulated by several protein kinases, including Cdk1 and Plk1. Several peripheral Golgi proteins that dissociate from the Golgi during mitosis are implicated in regulation of cytokinesis or chromosome segregation, thereby coordinating mitotic and cytokinetic events to Golgi rearrangement. Here we show that, at the onset of mitosis, Cdk1 phosphorylates the peripheral Golgi protein Nir2 at multiple sites; of these, S382 is the most prominent. Phosphorylation of Nir2 by Cdk1 facilitates its dissociation from the Golgi apparatus, and phospho-Nir2(pS382) is localized in the cleavage furrow and midbody during cytokinesis. Mitotic phosphorylation of Nir2 is required for docking of the phospho-Ser/Thr binding module, the Polo box domain of Plk1, and overexpression of a Nir2 mutant, which fails to interact with Plk1, affects the completion of cytokinesis. These results demonstrate a mechanism for coordinating mitotic and cytokinetic events with Golgi rearrangement during cell division. PMID- 15125836 TI - The carboxy-terminal extension of yeast ribosomal protein S14 is necessary for maturation of 43S preribosomes. AB - Eukaryotic ribosomal proteins are required for production of stable ribosome assembly intermediates and mature ribosomes, but more specific roles for these proteins in biogenesis of ribosomes are not known. Here we demonstrate a particular function for yeast ribosomal protein rpS14 in late steps of 40S ribosomal subunit maturation and pre-rRNA processing. Extraordinary amounts of 43S preribosomes containing 20S pre-rRNA accumulate in the cytoplasm of certain rps14 mutants. These mutations not only reveal a more precise function for rpS14 in ribosome biogenesis but also uncover a role in ribosome assembly for the extended tails found in many ribosomal proteins. These studies are one of the first to relate the structure of eukaryotic ribosomes to their assembly pathway the carboxy-terminal extension of rpS14 is located in the 40S subunit near the 3' end of 18S rRNA, consistent with a role for rpS14 in 3' end processing of 20S pre rRNA. PMID- 15125837 TI - Suppression of multiple substrate mutations by spliceosomal prp8 alleles suggests functional correlations with ribosomal ambiguity mutants. AB - Conformational change within the spliceosome is required between the first catalytic step of pre-mRNA splicing, when the branch site (BS) attacks the 5' splice site, and the second step, when the 5' exon attacks the 3' splice site, yielding mRNA and lariat-intron products. A genetic screen for suppressors of BS A-to-G mutants, which stall between the two steps, identified Prp8, the highly conserved spliceosomal factor. prp8 suppressors facilitate the second step for multiple intron mutants and interact functionally with first step suppressors, alleles of PRP16 and U6 snRNA. Genetic interactions among prp8, prp16, and U6 alleles suggest that these factors control a common stage in first-to-second step transition. We propose that mutant substrates are utilized by alteration of the equilibrium between first/second step conformations, resembling tRNA miscoding caused by altered equilibrium between open/closed ribosomal conformations. This mechanistic commonality suggests that alteration of rearrangements represents an evolutionarily convenient way of modulating substrate selectivity. PMID- 15125838 TI - Spontaneous sharp bending of double-stranded DNA. AB - Sharply bent DNA is essential for gene regulation in prokaryotes and is a major feature of eukaryotic nucleosomes and viruses. The explanation normally given for these phenomena is that specific proteins sharply bend DNA by application of large forces, while the DNA follows despite its intrinsic inflexibility. Here we show that DNAs that are 94 bp in length-comparable to sharply looped DNAs in vivo spontaneously bend into circles. Proteins can enhance the stability of such loops, but the loops occur spontaneously even in naked DNA. Random DNA sequences cyclize 10(2)-10(4) times more easily than predicted from current theories of DNA bending, while DNA sequences that position nucleosomes cyclize up to 10(5) times more easily. These unexpected results establish DNA as an active participant in the formation of looped regulatory complexes in vivo, and they point to a need for new theories of DNA bending. PMID- 15125839 TI - Structural basis for octameric ring formation and DNA interaction of the human homologous-pairing protein Dmc1. AB - The human Dmc1 protein, a RecA/Rad51 homolog, is a meiosis-specific DNA recombinase that catalyzes homologous pairing. RecA and Rad51 form helical filaments, while Dmc1 forms an octameric ring. In the present study, we crystallized the full-length human Dmc1 protein and solved the structure of the Dmc1 octameric ring. The monomeric structure of the Dmc1 protein closely resembled those of the human and archaeal Rad51 proteins. In addition to the polymerization motif that was previously identified in the Rad51 proteins, we found another hydrogen bonding interaction at the polymer interface, which could explain why Dmc1 forms stable octameric rings instead of helical filaments. Mutagenesis studies identified the inner and outer basic patches that are important for homologous pairing. The inner patch binds both single-stranded and double-stranded DNAs, while the outer one binds single-stranded DNA. Based on these results, we propose a model for the interaction of the Dmc1 rings with DNA. PMID- 15125840 TI - Involvement of transcription termination factor 2 in mitotic repression of transcription elongation. AB - All nuclear transcription is interrupted during mitosis. We examined the role of human TTF2, an RNA polymerase (Pol) I and II termination factor, in mitotic repression of transcription elongation. We find that TTF2 levels rise in the cytoplasm in S and G2 and at the onset of mitosis TTF2 translocates into the nucleus. Consistent with a role in termination of all transcription, TTF2 is the only ATP-dependent termination activity associated with Pol II transcription elongation complexes, is largely unaffected by template position, and is impervious to the phosphorylation state of the polymerase. Cells in which TTF2 levels are knocked down showed dramatic retention of Ser2 phosphorylated Pol II on mitotic chromosomes and an increase in chromosome segregation defects. PMID- 15125841 TI - Ssu72 Is an RNA polymerase II CTD phosphatase. AB - Phosphorylation of serine-2 (S2) and serine-5 (S5) of the C-terminal domain (CTD) of RNA polymerase II (RNAP II) is a dynamic process that regulates the transcription cycle and coordinates recruitment of RNA processing factors. The Fcp1 CTD phosphatase catalyzes dephosphorylation of S2-P. Here, we report that Ssu72, a component of the yeast cleavage/polyadenylation factor (CPF) complex, is a CTD phosphatase with specificity for S5-P. Ssu72 catalyzes CTD S5-P dephosphorylation in association with the Pta1 component of the CPF complex, although its essential role in 3' end processing is independent of catalytic activity. Depletion of Ssu72 impairs transcription in vitro, and this defect can be rescued by recombinant, catalytically active Ssu72. We propose that Ssu72 has a dual role in transcription, one as a CTD S5-P phosphatase that regenerates the initiation-competent, hypophosphorylated form of RNAP II and the other as a factor necessary for cleavage of pre-mRNA and efficient transcription termination. PMID- 15125843 TI - Structure of the BRCT repeats of BRCA1 bound to a BACH1 phosphopeptide: implications for signaling. AB - The recognition of the phosphorylated BACH1 helicase by the BRCA1 C-terminal (BRCT) repeats is important to the tumor suppressor function of BRCA1. Here we report the crystal structure of the BRCT repeats of human BRCA1 bound to a phosphorylated BACH1 peptide at 2.3 A resolution. The phosphorylated serine 990 and phenylalanine 993 of BACH1 anchor the binding to BRCA1 through specific interactions with a surface cleft at the junction of the two BRCT repeats. This surface cleft is highly conserved in BRCA1 across species, suggesting an evolutionarily conserved function of phosphopeptide recognition. Importantly, conserved amino acids critical for BACH1 binding are frequently targeted for missense mutations in breast cancer. These mutations greatly diminish the ability of BRCA1 to interact with the phosphorylated BACH1 peptide. Additional structural analysis revealed significant implications for understanding the function of the BRCT family of proteins in DNA damage and repair signaling. PMID- 15125842 TI - The IGF-1/PI3K/Akt pathway prevents expression of muscle atrophy-induced ubiquitin ligases by inhibiting FOXO transcription factors. AB - Skeletal muscle size depends upon a dynamic balance between anabolic (or hypertrophic) and catabolic (or atrophic) processes. Previously, no link between the molecular mediators of atrophy and hypertrophy had been reported. We demonstrate a hierarchy between the signals which mediate hypertrophy and those which mediate atrophy: the IGF-1/PI3K/Akt pathway, which has been shown to induce hypertrophy, prevents induction of requisite atrophy mediators, namely the muscle specific ubiquitin ligases MAFbx and MuRF1. Moreover, the mechanism for this inhibition involves Akt-mediated inhibition of the FoxO family of transcription factors; a mutant form of FOXO1, which prevents Akt phosphorylation, thereby prevents Akt-mediated inhibition of MuRF1 and MAFbx upregulation. Our study thus defines a previously uncharacterized function for Akt, which has important therapeutic relevance: Akt is not only capable of activating prosynthetic pathways, as previously demonstrated, but is simultaneously and dominantly able to suppress catabolic pathways, allowing it to prevent glucocorticoid and denervation-induced muscle atrophy. PMID- 15125844 TI - Strategies for successful advocacy. PMID- 15125845 TI - Effect of bracing on the quality of life of adolescents with idiopathic scoliosis. AB - BACKGROUND CONTEXT: There is considerable controversy regarding the effectiveness of bracing for patients with adolescent idiopathic scoliosis (AIS). Furthermore, little work has focused on the effect of formal bracing on the health-related quality of life (QOL) of affected adolescents. PURPOSE: This study is an attempt to assess the QOL of adolescents with AIS who were treated with a brace in comparison to those who were not. STUDY DESIGN/SETTING: This is a questionnaire based study of patients presenting to our institution for evaluation and treatment of AIS. The study was conducted at Columbia University and the New York Presbyterian Medical Center, New York, NY. PATIENT SAMPLE: A total of 136 patients with AIS who were being observed and 78 patients who were braced were included in this study. OUTCOME MEASURES: The Child Health Questionnaire (CHQ) and the American Academy of Orthopaedic Surgeons Pediatric Outcomes Data Collection Instrument (PODCI) were administered to 214 parents of adolescents undergoing evaluation for AIS. METHODS: Parents of AIS patients were administered the CHQ and PODCI to assess their child's QOL. Independent samples t tests were conducted to compare the QOL among braced versus observed patients. Univariate and multivariate analyses were conducted to determine the effect of gender, age, curvature and treatment type on QOL. RESULTS: There were few differences in QOL between braced versus observed patients, but boys tended to outscore the girls across most domains. Surprisingly, the QOL of adolescents undergoing bracing in this study was not significantly lower than age-adjusted norms. CONCLUSIONS: Brace wearing did not decrease the QOL of adolescents, compared with their observed counterparts, in our study population. PMID- 15125846 TI - Preemptive analgesia for postoperative pain relief in lumbosacral spine surgeries: a randomized controlled trial. AB - BACKGROUND CONTEXT: Administration of analgesic medication, before the actual onset of painful stimulus, is more effective than that after the onset of painful stimulus. This is the principle of preemptive analgesia. Although it is often considered superior to other forms of analgesia, its role in postoperative pain relief after lumbosacral spinal surgery has not been fully investigated. PURPOSE: To analyze the efficacy of preemptive analgesia with a single caudal epidural injection for patients undergoing surgeries on the lumbosacral spine by the posterior approach. STUDY DESIGN/SETTING: Randomized, double-blinded and controlled clinical trial. PATIENT SAMPLE: Eighty-two patients who underwent discectomy in the lumbosacral spine by the posterior approach, with or without instrumentation, were randomized to the control group (n=40) and to the study group (n=42). METHODS: Patients in control group received a single caudal epidural injection of 20 ml of normal saline. Patients in study group received a single caudal epidural injection of 20 ml containing bupivacaine and tramadol as the active agents. The time interval between this injection and the surgical incision was never less than 20 minutes in either of the groups. This facilitated enough time for the drug to get fixed to the nerve roots, leading to effective preemptive analgesia. OUTCOME MEASURES: Patients were monitored for postoperative pain immediately after surgery when they had completely recovered and regained consciousness from general anesthesia, and subsequently 4, 8, 12 and 24 hours thereafter. Pain was quantified using the visual analog scale (VAS) and the verbal rating scale (VRS). The time at which supplemental analgesic medication was first demanded in the postoperative period by the patient was also noted. RESULTS: The two groups were comparable for age, sex, body weight and the type of surgery they underwent. Because the data did not have a normal Gaussian distribution, the one-tailed Mann-Whitney test, being a nonparametric test, was adopted for statistical analysis. Accordingly, VAS and VRS values at all time intervals were significantly lower (p<.0001) in the study group as compared with the control group. This indicated significantly better pain relief in the study group. There was also a significant delay (p=.0041) in the first demand for supplemental analgesic medication in the postoperative period in the study group. No complication specific to the procedure was noted except for the development of postoperative urinary retention, which was transient and appropriately managed with urinary catheterization. CONCLUSIONS: Preemptive analgesia with a single caudal epidural injection of bupivacaine and tramadol is a safe, simple and effective method for postoperative pain relief. PMID- 15125847 TI - Stiffness of prosthetic nucleus determines stiffness of reconstructed lumbar calf disc. AB - BACKGROUND CONTEXT: Currently, artificial spinal discs require transection or partial removal of the annulus fibrosis in order to excise the nucleus and implant a prosthetic nucleus or implant a total disc device, respectively. Preservation of the annulus for prosthetic disc replacement maintains the function of the annulus and may improve annulus load sharing with the prosthesis. PURPOSE: To quantify the biomechanical characteristics of an annular sparing intervertebral prosthetic disc (IPD) in a lumbar calf spine model. The aim of the study was to determine whether altering the stiffness of the elastic component of this unique prosthesis would correspond to changes of the overall reconstructed disc. STUDY DESIGN/SETTING: A biomechanical study was conducted in vitro using cadaveric calf spines such that each specimen served as its own control. Investigations were performed at the Minneapolis Medical Research Foundation, Orthopaedic Biomechanics Laboratory. METHODS: Six L45 or L56 motion segments (from which the posterior elements had been removed) were studied in axial compression, sagittal and lateral bending and torsion. These load states were applied to the intact, denucleated and prosthetically reconstructed disc using four IPDs of differing stiffness. RESULTS: Load-displacement testing demonstrated that stiffer IPDs resulted in a decreased range of motion and neutral zone, and greater stiffness of the reconstructed disc. Disc reconstruction with the stiffest IPD approximated the behavior of the intact disc. CONCLUSIONS: The overall biomechanical characteristics of a reconstructed disc are related to the stiffness of a nucleus prosthesis. The similarities in the mechanical behavior of reconstructed and intact discs suggest that additional feasibility studies for the annulus-sparing IPD are warranted. PMID- 15125848 TI - Sensory innervation of the lateral portion of the lumbar intervertebral disc in rats. AB - BACKGROUND CONTEXT: An annular tear extending to the outer one-third of the annulus is thought to be one of the causes of low back pain. However, some patients have bilateral low back symptoms, even if the annular tear is localized in the lateral disc. Because nociceptive information from the lateral disc is transmitted by the dorsal root ganglion (DRG) neurons innervating the lateral disc, we investigated the distribution of the DRG neurons innervating the lateral portion of the disc. PURPOSE: To clarify the distribution and pathway of the DRG neurons innervating the lateral portion of the L5-L6 disc in rats. STUDY DESIGN/SETTING: Using the retrograde tracing method, we studied the innervation pattern of the lateral portion of the L5-L6 intervertebral disc in rats. METHODS: The retrograde transport of Fluoro-Gold (F-G; Fluorochrome, Denver, CO) was used in 22 rats. Subjects included a nontreated group (n=16) and a sympathectomized group (n=6). Seven days after the application of F-G crystals to the left lateral portion of the L5-L6 disc, bilateral T12-L6 DRGs were observed by fluorescent microscopy. RESULTS: In the nontreated group, of all the F-G-labeled neurons, 93.1% were present in the left DRGs and 6.9% were in the right DRGs. The number of labeled neurons was largest in the left L2 DRGs. In the sympathectomized group, the numbers of labeled neurons in the T13, L1 and L2 DRGs were significantly lower than the numbers in the nontreated group. CONCLUSION: Results of this study indicate that DRG neurons innervating the lateral portion of the disc are distributed mainly in the ipsilateral side but also in the contralateral side. The DRG neurons in T13, L1 and L2 innervate the lateral portion of the L5 L6 disc through the paravertebral sympathetic trunks. PMID- 15125850 TI - Posterolateral lumbar fusions in athymic rats: characterization of a model. AB - BACKGROUND CONTEXT: The athymic rat has been used to study the role of osteoinductive products in spinal fusions. This small animal model has been advocated to minimize potential inflammatory responses to allogeneic or xenogenic proteins. Despite past experience, this model has not yet been well characterized. PURPOSE: To further define and validate a posterolateral lumbar fusion model in the athymic rat. STUDY DESIGN/SETTING: Comparison of fusions after animal survival surgery. PATIENT SAMPLE: Forty athymic and 20 normothymic rats. OUTCOME MEASURES: Manual palpation, radiography and histology at 3 and 6 weeks. METHODS: Single-level intertransverse fusions were performed at the L4-L5 level of 40 athymic rats. Twenty rats were implanted with autograft (athymic/autograft), and 20 had no graft placed (athymic/no graft). An additional 20 autograft fusions were performed on normothymic rats (normothymic/autograft). Half were sacrificed at 3 weeks; half were sacrificed at 6 weeks. RESULTS: At 3 weeks, 0% of the athymic/no graft rats fused, 20% of the athymic/autograft rats fused and 20% of the normothymic/autograft rats fused by manual palpation. At 6 weeks, 0% of the athymic/no graft rats fused, 30% of the athymic/autograft rats fused and 40% of the normothymic/autograft rats fused by manual palpation. Radiographs were of limited utility in determining fusion, and histology results were roughly concordant with those of manual palpation. CONCLUSIONS: This work further characterizes the athymic rat posterolateral lumbar fusion model. The absence of a thymus does not appear to affect autograft fusion rates, and no spontaneous fusions were seen when no graft was placed. PMID- 15125851 TI - Posterolateral intertransverse lumbar arthrodesis in the New Zealand White rabbit model: I. Surgical anatomy. AB - BACKGROUND CONTEXT: The New Zealand White rabbit model for posterolateral lumbar fusion is commonly used for spinal fusion research. However, the high rate of animal morbidity and mortality associated with the model makes experimentation inefficient and can lead to faulty data analysis. Operative complications are in part the result of inadequate knowledge of normal rabbit lumbar spine anatomy. PURPOSE: To describe the lumbar spine anatomy of the New Zealand White rabbit as it pertains to the surgical technique of posterolateral intertransverse arthrodesis. STUDY DESIGN/SETTING: This is a descriptive anatomical study of the lumbar spine (and related structures) of the New Zealand White rabbit spinal fusion model. The study was performed at a university research facility. METHODS: The lumbar spine and associated soft tissue structures of 16 previously sacrificed, skeletally mature New Zealand White rabbits were dissected and examined. RESULTS: The musculoskeletal and neurologic structures relevant to posterolateral lumbar fusion in the New Zealand White rabbit are described. CONCLUSIONS: Specific knowledge of rabbit lumbar spine anatomy allows the researcher to more rapidly acquire expertise in the surgical technique of posterolateral arthrodesis. Improved technical execution of the procedure should lower the complication rate, reduce the costs of experimentation and lead to more reliable and reproducible results. PMID- 15125852 TI - Posterolateral intertransverse lumbar arthrodesis in the New Zealand White rabbit model: II. Operative technique. AB - BACKGROUND CONTEXT: Although the New Zealand White (NZW) rabbit model has been increasingly used for spinal fusion research, reported complication rates have been as high as 23%. The substantial animal morbidity and mortality associated with the model makes experimentation inefficient and can lead to faulty data analysis. Operative complications are in part the result of suboptimal execution of the experimental protocol. PURPOSE: To describe an experimental protocol for posterolateral intertransverse lumbar arthrodesis in the NZW rabbit and to analyze the effect of the protocol on the complication rate of this model. STUDY DESIGN/SETTING: This is a descriptive analysis of the surgical technique (and related complications) used for posterolateral spinal arthrodesis in the NZW rabbit model. This study was performed at a university research facility. METHODS: The complications of posterolateral intertransverse lumbar arthrodesis in the NZW rabbit were analyzed before and after instituting a refined experimental protocol. RESULTS: A total of 77 NZW rabbits underwent a posterolateral lumbar arthrodesis. In the first 48 animals (Group A) complications occurred at a rate of 26% and included anesthetic demise, nerve palsies, wound infections or seromas and fusion of the wrong level. After the institution of a refined experimental protocol, there were no complications in a second group of 29 rabbits (Group B). CONCLUSIONS: Adherence to a precise experimental protocol for posterolateral lumbar arthrodesis can minimize complications, thereby enhancing the NZW rabbit as a cost-effective and accurate animal model for spinal fusion research. PMID- 15125854 TI - Direct current stimulation of titanium interbody fusion devices in primates. AB - BACKGROUND CONTEXT: The fusion rate for anterior lumbar interbody fusion (ALIF) varies widely with the use of different interbody devices and bone graft options. Adjunctive techniques such as electrical stimulation may improve the rate of bony fusion. PURPOSE: To determine if direct current (DC) electrical stimulation of a metallic interbody fusion device enhanced the incidence or extent of anterior bony fusion. STUDY DESIGN/SETTING: ALIF was performed using titanium alloy interbody fusion devices with and without adjunctive DC electrical stimulation in nonhuman primates. METHODS: ALIF was performed through an anterolateral approach in 35 macaques with autogenous bone graft and either a titanium alloy (Ti-6Al-4V) fusion device or femoral allograft ring. The fusion devices of 19 animals received high (current density 19.6 microA/cm2) or low (current density 5.4 microA/cm2) DC electrical stimulation using an implanted generator for a 12- or 26-week evaluation period. Fusion sites were studied using serial radiographs, computed tomography imaging, nondestructive mechanical testing and qualitative and semiquantitative histology. RESULTS: Fusion was achieved with the titanium fusion device and autogenous bone graft. At 12 weeks, the graft was consolidating and early to moderate bridging callus was observed in and around the device. By 26 weeks, the anterior callus formation was more advanced with increased evidence of bridging trabeculations and early bone remodeling. The callus formation was not as advanced or abundant for the allograft ring group. Histology revealed the spinal fusion device had an 86% incidence of bony fusion at 26 weeks compared with a 50% fusion rate for the allograft rings. DC electrical stimulation of the fusion device had a positive effect on anterior interbody fusion by increasing both the presence and extent of bony fusion in a current density-dependent manner. CONCLUSIONS: Adjunctive DC electrical stimulation of the fusion device improved the rate and extent of bony fusion compared with a nonstimulated device. The fusion device was equivalent to or better than the femoral allograft ring in all evaluations. The use of adjunctive direct current electrical stimulation may provide a means of improving anterior interbody fusion. PMID- 15125855 TI - Pulmonary complications in anterior-posterior thoracic lumbar fusions. AB - BACKGROUND CONTEXT: Surgery for adult spinal deformity may require both an anterior and posterior approach in order to stabilize the spine and achieve the desired correction. These procedures can be associated with significant pulmonary complications, including atelectasis, pneumonia and respiratory failure. The etiology of some of the respiratory complications is clear: poor inspiratory effort from incision pain and previous pulmonary disease. However, for many patients the direct cause of these complications is not obvious. PURPOSE: To delineate the incidence, severity and risks associated with pulmonary complications in the setting of major spine surgery. STUDY DESIGN/SETTING: Retrospective chart review study of adult patients undergoing combined anterior posterior thoracic, lumbar and sacral fusion spine surgery. PATIENT SAMPLE: A total of 60 charts were reviewed for this study. OUTCOME MEASURES: Radiographic abnormalities correlated with clinical findings, postoperative need for ventilation and lengths of hospital stay were used as outcome measures. METHODS: Perioperative pulmonary complications were assessed for 60 patients with spinal deformities who underwent combined anterior-posterior thoracic, lumbar and sacral fusion over a 2-year period. RESULTS: One patient was eliminated from analysis because of multiple surgeries during his hospital course. Of the remaining 59 patients, 38 (64%) developed roentgenographic abnormalities. The most common radiographic finding was an effusion found in 66% of these patients, followed by atelectasis in 53%. Twenty-one percent (8 of 38) had infiltrates. Five (5 of 38) or 13% had evidence of partial or complete lobar collapse; in two bronchoscopy was required because of profound hypoxemia. Two patients had pneumonia requiring antibiotic treatment. All but two patients were extubated within 36 hours of surgery. They were kept intubated because of hemodynamic instability. There was no statistically significant difference in the group of patients with and without roentgenographic abnormalities with regard to age, weight, American Society of Anesthesiologists class, smoking history, pulmonary function test results, blood loss, perioperative blood and crystalloid requirement and length of surgery. Patients with radiographic abnormalities were more likely to have had invasion of their thoracic cavity (p=.02) and had a longer mean hospital stay of 13.5 versus 10.2 days (p=.009). CONCLUSION: Radiographic abnormalities of the lungs are common after major spine surgery involving both an anterior and posterior approach, especially when the thoracic cavity is invaded. In view of the morbidity and longer hospital stay associated with such findings, close monitoring of pulmonary status with aggressive pulmonary toilet are indicated. PMID- 15125857 TI - Thoracoscopic transdiaphragmatic approach to thoracolumbar junction fractures. AB - BACKGROUND CONTEXT: Anterior approaches to the thoracocolmbar junction (TLJ) are often required to restore anterior column deficiency after spinal trauma. Conventional open approaches are often associated with significant morbidity, and hence there is a need for a minimally invasive approach to TLJ fractures. PURPOSE: To report the feasibility and effectiveness of the thoracoscopic transdiaphragmatic approach (TTA) in the management of TLJ fractures. STUDY DESIGN: A retrospective analysis of 212 patients undergoing surgery at two institutions by the TTA with neurological outcomes, fusion rates and complications. PATIENT SAMPLE: This is a two-institution study of 212 patients managed by TTA, from Berufsgenossenschaftliche Unfallklinik Marnau, a regional trauma facility located in Murnau, Bavaria, Germany, and from Stanford University, Stanford, California from May 1996 to June 2002. Patient ages ranged from 16 to 75 years (mean, 36 years) and included 158 males and 62 females. OUTCOME MEASURES: The neurological status was assessed by the Frankel Neurological Performance scale pre- and postoperatively. Plain radiographs obtained 1 year postoperatively assessed fusion radiologically. METHODS: All patients underwent spinal decompression, reconstruction and instrumentation by the TTA. Seventy-five patients had anterior instrumentation alone, whereas the remaining 137 had combined anterior and posterior instrumentation. A Z-Plate was used for spinal instrumentation from May 1996 to October 1999 and the MACS-TL system from November 1999 to June 2002. RESULTS: Monosegmental, bisegmental and multisegmental fixations were used in 46%, 48% and 6% of cases, respectively. Follow-up ranged from 12 months to 6 years (mean, 3.9 years). Surgical durations ranged between 70 minutes and 7 hours (mean, 3.5 hours). Successful bony fusion with maintenance of satisfactory spinal alignment was observed in approximately 90% of our patients. Anterior screw loosening was seen in five cases (2.4%), four involving the Z-Plate system and the other involving the MACS-TL system. Three patients (1.4%) required conversion to an open procedure. Access-related complications, such as pleural effusion, pneumothorax and intercostal neuralgia, were seen in 12 patients (5.7%). Three patients (1.4%) had superficial portal infections. We encountered no diaphragmatic herniations. CONCLUSIONS: TTA provides excellent access to the entire TLJ, permitting satisfactory spinal decompression, reconstruction and instrumentation. Diaphragmatic opening and repair can be accomplished safely and effectively without special endoscopic instrumentation. It also precludes the need for retroperitoneoscopic or open thoracoabdominal approaches and thus avoids the associated significant morbidity. PMID- 15125859 TI - Rheumatoid arthritis of the cervical spine. AB - BACKGROUND CONTEXT: Rheumatoid arthritis affects over 2 million patients in the United States. It is the most common inflammatory disorder of the cervical spine. The natural history is variable. Women tend to be more commonly involved than men. Atlantoaxial instability is the most common form of cervical involvement and may occur either independently or concomitantly with cranial settling and subaxial instability. Cervical spine involvement can be seen in up to 86% of patients and neurologic involvement in up to 58%. Myelopathy is rare but when present portends a poor prognosis. What is frustrating for clinicians treating these patients is that pain cannot be equated with instability or instability with neurologic symptoms. The goal is to identify patients at risk before the development of neurologic symptoms. Both radiographic and nonradiographic risk factors play an important role in the surgical decision-making process. PURPOSE: We will describe the current concepts in rheumatoid arthritis of the cervical spine. Emphasis is placed on the natural history, anatomy, pathophysiology and decision-making process. STUDY DESIGN: A review of the current concepts of rheumatoid arthritis of the cervical spine. METHODS: MEDLINE search of all English literature published on rheumatoid arthritis of the cervical spine. RESULTS: Rheumatoid arthritis of the cervical spine was first described by Garrod in 1890. The prevalence has been estimated to be 1% to 2% of the world's adult population. Despite its prevalence, the etiology of rheumatoid arthritis remains unknown. Because of its potentially debilitating and life-threatening sequelae in advanced disease, rheumatoid arthritis in the cervical spine today remains a high priority to diagnose and treat. CONCLUSIONS: Many aspects of the natural history and pathophysiology of the rheumatoid spine remain unclear. The timing of operative intervention in patients with radiographic instability and no evidence of neurologic deficit is an area of considerable controversy. Continued surveillance into the natural history of the rheumatoid spine is required. PMID- 15125860 TI - Efficacy of spinal manipulation and mobilization for low back pain and neck pain: a systematic review and best evidence synthesis. AB - BACKGROUND CONTEXT: Despite the many published randomized clinical trials (RCTs), a substantial number of reviews and several national clinical guidelines, much controversy still remains regarding the evidence for or against efficacy of spinal manipulation for low back pain and neck pain. PURPOSE: To reassess the efficacy of spinal manipulative therapy (SMT) and mobilization (MOB) for the management of low back pain (LBP) and neck pain (NP), with special attention to applying more stringent criteria for study admissibility into evidence and for isolating the effect of SMT and/or MOB. STUDY DESIGN: RCTs including 10 or more subjects per group receiving SMT or MOB and using patient-oriented primary outcome measures (eg, patient-rated pain, disability, global improvement and recovery time). METHODS: Articles in English, Danish, Swedish, Norwegian and Dutch reporting on randomized trials were identified by a comprehensive search of computerized and bibliographic literature databases up to the end of 2002. Two reviewers independently abstracted data and assessed study quality according to eight explicit criteria. A best evidence synthesis incorporating explicit, detailed information about outcome measures and interventions was used to evaluate treatment efficacy. The strength of evidence was assessed by a classification system that incorporated study validity and statistical significance of study results. Sixty-nine RCTs met the study selection criteria and were reviewed and assigned validity scores varying from 6 to 81 on a scale of 0 to 100. Forty-three RCTs met the admissibility criteria for evidence. RESULTS: Acute LBP: There is moderate evidence that SMT provides more short-term pain relief than MOB and detuned diathermy, and limited evidence of faster recovery than a commonly used physical therapy treatment strategy. Chronic LBP: There is moderate evidence that SMT has an effect similar to an efficacious prescription nonsteroidal anti-inflammatory drug, SMT/MOB is effective in the short term when compared with placebo and general practitioner care, and in the long term compared to physical therapy. There is limited to moderate evidence that SMT is better than physical therapy and home back exercise in both the short and long term. There is limited evidence that SMT is superior to sham SMT in the short term and superior to chemonucleolysis for disc herniation in the short term. However, there is also limited evidence that MOB is inferior to back exercise after disc herniation surgery. Mix of acute and chronic LBP: SMT/MOB provides either similar or better pain outcomes in the short and long term when compared with placebo and with other treatments, such as McKenzie therapy, medical care, management by physical therapists, soft tissue treatment and back school. Acute NP: There are few studies, and the evidence is currently inconclusive. Chronic NP: There is moderate evidence that SMT/MOB is superior to general practitioner management for short-term pain reduction but that SMT offers at most similar pain relief to high-technology rehabilitative exercise in the short and long term. Mix of acute and chronic NP: The overall evidence is not clear. There is moderate evidence that MOB is superior to physical therapy and family physician care, and similar to SMT in both the short and long term. There is limited evidence that SMT, in both the short and long term, is inferior to physical therapy. CONCLUSIONS: Our data synthesis suggests that recommendations can be made with some confidence regarding the use of SMT and/or MOB as a viable option for the treatment of both low back pain and NP. There have been few high-quality trials distinguishing between acute and chronic patients, and most are limited to shorter-term follow-up. Future trials should examine well-defined subgroups of patients, further address the value of SMT and MOB for acute patients, establish optimal number of treatment visits and consider the cost-effectiveness of care. PMID- 15125861 TI - Cystic lymphangiomas of the cauda equina. AB - BACKGROUND CONTEXT: Lymphangiomas, benign hamartomatous lesions of the lymph tissue, result from a failure of lymph channels to communicate with the venous system or normal lymph channels. Previously, two reports described lymphangiomas arising in the peripheral nerve (ulnar nerve and posterior tibial nerve). PURPOSE: To report a rare case of cystic lymphangiomas of the cauda equina not previously reported. STUDY DESIGN: Case report. PATIENT SAMPLE: A 56-year-old Japanese man. METHODS: Not applicable. RESULT: The patient recovered from most of his symptoms after tumor extirpation. He had no symptoms at a 5.8-year follow-up. CONCLUSIONS: We believe this is the first case report of lymphangiomas originating from the cauda equina, which was effectively treated by surgery. PMID- 15125862 TI - Use of somatosensory evoked potentials to detect peripheral ischemia and potential injury resulting from positioning of the surgical patient: case reports and discussion. AB - BACKGROUND: Somatosensory evoked potentials (SSEPs) have long been recognized as an excellent tool for detecting neural and vascular compromise during vascular, neurosurgical and orthopedic procedures. SSEPs have the ability to localize, central versus peripheral, the area of compromise. Many surgeons use only lower limb SSEP monitoring when performing lumbar spinal surgery. The upper extremities are usually not monitored during such procedures, and monitoring oxygen saturation does not detect neural compromise. PURPOSE: To report that the expanded use of SSEP monitoring during surgery can be beneficial in detecting peripheral ischemia or neural compromise resulting from positioning. STUDY DESIGN: Three case reviews of orthopedic spine surgeries where SSEP monitoring provided early warnings of vascular and neural compression. METHODS: The cases review three different lumbar procedures in which evidence of peripheral ischemia and nerve compression were detected by SSEP monitoring. RESULTS: By the use of upper- and lower-extremity monitoring during lumbar procedures, early detection of ischemia and nerve compression were noted intraoperatively. These changes prompted examination of the patient and repositioning to correct the ischemia or compression. The repositioning in these cases corrected the problem, and no lasting effects were found. CONCLUSIONS: Including SSEP monitoring of the bilateral upper extremities should be considered during lumbar spinal procedures. Such monitoring can be offered for a slightly increased expense and only minimal time delay to place the additional required electrodes by the technician. As a direct result of the early warning of the SSEP monitoring, we were able to avoid potential ischemic injuries and improve patient outcomes. PMID- 15125864 TI - Paraplegia after lumbosacral nerve root block: report of three cases. PMID- 15125866 TI - Planning for labor (and labor analgesia) in a parturient with spinal cord injury: a need for multidisciplinary approach. PMID- 15125867 TI - Antiviral drugs in current clinical use. AB - The current armamentarium for the chemotherapy of viral infections consists of 37 licensed antiviral drugs. For the treatment of human immunodeficiency virus (HIV) infections, 19 compounds have been formally approved: (i) the nucleoside reverse transcriptase inhibitors (NRTIs) zidovudine, didanosine, zalcitabine, stavudine, lamivudine, abacavir and emtricitabine; (ii) the nucleotide reverse transcriptase inhibitor (NtRTI) tenofovir disoproxil fumarate; (iii) the non-nucleoside reverse transcriptase inhibitors (NNRTIs) nevirapine, delavirdine and efavirenz; (iv) the protease inhibitors saquinavir, ritonavir, indinavir, nelfinavir, amprenavir, lopinavir (combined with ritonavir at a 4/1 ratio) and atazanavir; and the viral entry inhibitor enfuvirtide. For the treatment of chronic hepatitis B virus (HBV) infections, lamivudine as well as adefovir dipivoxil have been approved. Among the anti-herpesvirus agents, acyclovir, valaciclovir, penciclovir (when applied topically), famciclovir, idoxuridine and trifluridine (both applied topically) as well as brivudin are used in the treatment of herpes simplex virus (HSV) and/or varicella-zoster virus (VZV) infections; and ganciclovir, valganciclovir, foscarnet, cidofovir and fomivirsen (the latter upon intravitreal injection) have proven useful in the treatment of cytomegalovirus (CMV) infections in immunosuppressed patients (i.e. AIDS patients with CMV retinitis). Following amantadine and rimantadine, the neuraminidase inhibitors zanamivir and oseltamivir have recently become available for the therapy (and prophylaxis) of influenza virus infections. Ribavirin has been used (topically, as aerosol) in the treatment of respiratory syncytial virus (RSV) infections, and the combination of ribavirin with (pegylated) interferon-alpha has received increased acceptance for the treatment of hepatitis C virus (HCV) infections. PMID- 15125868 TI - A new primer set improves the efficiency of competitive PCR-ELISA for the detection of B19 DNA. PMID- 15125869 TI - Human herpesvirus 8 seroconversion in Kenyan women by enzyme-linked immunosorbent assay and immunofluorescence assay. AB - BACKGROUND: Human herpesvirus 8 (HHV-8) antibody tests vary in reported sensitivity and specificity, depending on the population tested and the assay. OBJECTIVE: The purpose of this study was to compare the ability to detect seroconversion to HHV-8 in a cohort of HHV-8 seronegative female commercial sex workers in Kenya using three tests: HHV-8 viral lysate-based enzyme-linked immunosorbent assay (ELISA), an immunofluorescence assay for HHV-8 lytic antigens (IFA-lytic) and IFA for latent nuclear antigens (IFA-LANA). STUDY DESIGN: By ELISA, 16 women from a prospective cohort of commercial sex workers were identified as seroconverting to HHV-8. A total of 124 post-enrollment samples from these 16 women as well as the enrollment samples were tested for HHV-8 antibodies by all three assays to monitor seroconversion. RESULTS: Of 16 women with apparent seroconversion by ELISA, 8 had a rise in IFA-lytic titers either concomitant with or prior to the first positive ELISA sample and no initial LANA by IFA. Five of the 16 women were IFA-LANA positive at entry, indicating prior infection with HHV-8. Three women had no evidence of seroconversion by either IFA lytic or IFA-LANA and two of these three had increased ELISA reactivity concomitant with HIV-1 infection. CONCLUSIONS: Conversion from a negative to a positive ELISA result for HHV-8 antibody indicated seroconversion in only half of the study cohort of 16 women when IFA-lytic and IFA-LANA results were considered. The IFA-lytic assay was more sensitive than ELISA for early antibody responses. The IFA-LANA was positive in some women who had neither IFA-lytic nor ELISA antibodies suggesting it may be a marker for latent infections. Presumptive identification of incident HHV-8 infection by ELISA screening followed by IFA lytic testing to confirm the positive test and IFA-LANA to rule out prior infection provides the most accurate documentation of HHV-8 seroconversion. PMID- 15125870 TI - Epidemiology of HHV-8 infection in HIV-positive patients with and without Kaposi sarcoma: diagnostic relevance of serology and PCR. AB - BACKGROUND: Kaposi's sarcoma (KS) remains the most common neoplasm in HIV infected patients. Human herpesvirus 8 (HHV-8) infection is etiologically associated with KS. Diagnostic procedures with regard to HHV-8 infection are not routinely performed in HIV-infected patients; diagnostic and prognostic value of HHV-8 serology or PCR are unknown in this setting. Epidemiological data concerning HHV-8 infection of HIV-infected patients in Germany are rare. OBJECTIVES: To assess prevalence of HHV-8 infection in a cohort of HIV-infected patients with and without KS in Germany and to correlate this to manifestations and clinical course of KS. STUDY DESIGN: HHV-8 serology was performed in 483 patients in routine care for HIV-infection in northern Germany. HHV-8 DNA was analyzed by PCR in peripheral blood mononuclear cells (PBMC) of 293 patients; in a subgroup multiple samples were analyzed. History and manifestations of KS were recorded. RESULTS: HHV-8 antibodies were detected using IFT in 91% of 33 patients with KS and 52% of 398 patients without KS. In 36 of 293 (12.3%) patients HHV-8 DNA was detected in PBMC. In general, HHV-8 DNA was not continuously detected when multiple samples from the same patient were analyzed. Patients with KS history were more likely to be PCR positive than those without (45.5% versus 7.8%). In patients with active KS HHV-8 DNA was detected more frequently than in patients with disease remission. HHV-8 DNA was not detected in serologically negative patients. However, three patients with KS history in full remission for several years were seronegative. CONCLUSIONS: HIV-infected patients were frequently found to be positive for HHV-8 antibodies. The number of patients positive for viral DNA in PBMC was much smaller. Single PCR-examinations were of little value for prognosis, but repeated detection of HHV-8 DNA represents an increased risk of disease activity. PMID- 15125871 TI - A sensitive and quantitative single-tube real-time reverse transcriptase-PCR for detection of enteroviral RNA. AB - BACKGROUND: Enteroviruses (EVs) are significant human pathogens. Rapid and sensitive diagnostic techniques are desirable. OBJECTIVES: To develop a quantitative single-tube real-time reverse transcription-polymerase chain reaction (RT-PCR) for human enterovirus ribonucleic acid (RNA) (QPCR), with protection against amplimer contamination. STUDY DESIGN: The method was evaluated with serial dilutions of EV, 62 cerebrospinal fluid (CSF) specimens from meningitis patients, and the third and fourth European Union Concerted Action Enterovirus Proficiency Panels. A commercial EV PCR test was run in parallel. RESULTS: Optimisation included RNA extraction procedure, design and concentrations of primers and probes from the 5' non-coding region as well as recombinant Thermus thermophilus polymerase (rTth), Mn(OAc)(2) and thermolabile UNG concentrations. Of 62 CSF samples from cases of meningitis submitted for QPCR testing, 34 (76%) and 21 (47%) were positive by QPCR and a commercial EV RNA detection kit, respectively. The detection limit of QPCR was 0.001 TCID(50)/ml (50% tissue culture-infective dose per millilitre) for a coxsackievirus B2 preparation and <10 copies of a plasmid containing coxsackievirus B2 complementary deoxyribonucleic acid (cDNA). The relation between threshold cycle (C(t)) and amount of virus was linear (r = 0.99) over a range of 10(-3) to 10(4) TCID(50)/ml of coxsackievirus B2. CONCLUSIONS: The QPCR method allows a large number of samples to be screened rapidly. Its sensitivity, simplicity, and reproducibility make it a suitable tool for the routine laboratory. PMID- 15125872 TI - Assessment of automated DNA extraction coupled with real-time PCR for measuring Epstein-Barr virus load in whole blood, peripheral mononuclear cells and plasma. AB - BACKGROUND: Epstein-Barr virus (EBV) DNA load monitoring in blood has been shown to be essential for the diagnosis of EBV-associated diseases. However, the methods currently used to assess EBV DNA load are often time-consuming and require prior blood separation. OBJECTIVES: The aim of this study was to evaluate the relative diagnostic value of EBV DNA load monitoring in whole blood, peripheral blood mononuclear cells (PBMCs) and plasma after automated DNA extraction using the MagNA Pure extractor followed by LightCycler real-time quantitative PCR (LC-PCR). STUDY DESIGN: First, EBV DNA load was assessed retrospectively after automated or manual extraction on 104 PBMC specimens. Second, EBV DNA load was determined prospectively with the automated extraction procedure in the whole blood, PBMCs and plasma of 100 samples from patients with EBV-related diseases (group 1, n = 20), HIV-seropositive individuals (group 2, n = 66), and healthy EBV carriers (group 3, n = 14). RESULTS: A good correlation was observed between automated and manual extraction on 104 PBMC specimens (r = 0.956; P < 0.0001). In the prospective study, 67 samples were positive in both whole blood and PBMCs, with a good correlation between EBV DNA loads in whole blood and PBMCs (r = 0.936; P < 0.0001). Only 18/100 samples were positive in plasma. Higher viral loads were regularly observed in the three blood compartments from group 1 than from groups 2 and 3. CONCLUSION: This study demonstrated that an automated extraction of EBV DNA is easier to perform in whole blood or plasma than in PBMCs and facilitates the standardisation of EBV DNA measurement by real-time quantitative PCR. The quantitative detection of EBV DNA load in whole blood appeared more sensitive than in plasma for infectious mononucleosis in immunocompetent patients, probably because of a rapid loss of plasmatic EBV DNA. In transplant patients, EBV DNA load monitoring in whole blood and in plasma turned out to be equivalent in terms of feasibility and accuracy for the early diagnosis of post-transplant lymphoproliferative diseases (PTLDs). PMID- 15125873 TI - Evaluation of a multiplex reverse transcriptase PCR ELISA for the detection of nine respiratory tract pathogens. AB - BACKGROUND: A multiplex reverse transcription (RT) polymerase chain reaction combined with a microwell hybridization assay (m-RT-PCR-ELISA) was previously developed to detect nine different microorganisms: enterovirus (EV), influenza virus type A (IVA) and type B (IVB), respiratory syncytial virus (RSV), parainfluenzavirus type 1 (PIV1) and type 3 (PIV3), adenovirus (AV), Mycoplasma pneumoniae (Mpn), Chlamydia pneumoniae (Cpn) in a single test. These organisms do not usually colonize the respiratory tract of humans, but, if present, it may be assumed they are involved in respiratory disease. OBJECTIVES AND STUDY DESIGN: The m-RT-PCR-ELISA was tested on (i) culture supernatants of unknown contents, (ii) by determining the analytical sensitivity of 10-fold serial dilutions of culture supernatants and (iii) by determining clinical sensitivity in a retrospective study on 411 clinical specimens. The specimens were re-tested in parallel by m-RT-PCR-ELISA versus the gold standard culture and immunfluorescence, and versus individual RT-PCR. RESULTS: (i) The 9-valent m-RT PCR-ELISA shows 83% to 100% concordant results on 103 culture supernatants containing different organisms. (ii) The analytical sensitivity was as follows: higher sensitivity of the 9-valent m-RT-PCR-ELISA in comparison to culture in the cases of PIV3, IVA and IVB (factor 10) and AV and EV (factor 100), and lower sensitivity in case of RSV and PIV1 (factor 10). (iii) The agreement with the gold standard in the kappa statistic was excellent for RSV (kappa = 0.937), IVA (kappa = 0.940), very good for PIV1 (kappa = 0.914), IVB (kappa = 0.907) and satisfactory for PIV3 (kappa = 0.410). For AV, EV and Mpn the m-RT-PCR-ELISA preliminary could be qualified as very good, based on the data derived on culture supernatants. Information about the validity for Cpn is limited. CONCLUSION: The m-RT-PCR-ELISA is a feasible, sensitive and specific method for detection of a broad spectrum of organisms. It is suitable for individual as well as epidemiological diagnosis. PMID- 15125874 TI - Improved detection of HCV Infection in hemodialysis patients using a new HCV RNA qualitative assay: experience of a transplant center. AB - BACKGROUND: Hepatitis C virus (HCV) is frequently a silent infection in hemodialysis (HD) patients with a prevalence of 8-10%. Improving HCV detection in this population prior to transplantation is critical both for infection control and optimal patient care. OBJECTIVES: To assess the current HCV testing practice of the National Institute for Transplantation (PCR testing of enzyme immunoassay (EIA) positive HD patients) by evaluating a subset of EIA positive and EIA negative samples with the VERSANT HCV RNA Qualitative Assay based on transcription mediated amplification (HCV Qual (TMA)) (sensitivity < or = 9.6 IU/ml) and in-house PCR (HCV Qual (PCR)) (sensitivity approximately 149 IU/ml). STUDY DESIGN: 2321 HD patients were screened by Abbott HCV EIA 2.0. A subset of 80/169 E IA positive samples and 100/2152 EIA negative samples were tested by both assays. TMA/PCR discordant samples were genotyped. RESULTS: PCR and TMA gave concordant results in 67/80 (83.8%) of EIA positive samples. 11/80 (14.7%) were reactive by HCV Qual (TMA), but not by HCV Qual (PCR); 2/80 (2.7%) were reactive by HCV Qual (PCR), but not by HCV Qual (TMA). 2/100 (2%) EIA negative samples were reactive and 95/100 (95%) were non-reactive by both assays. Three (3%) were only HCV Qual (TMA) reactive. 11/14 TMA+/PCR-samples with sufficient volume were genotyped. CONCLUSIONS: HCV Qual (TMA) identified active HCV infection in more EIA positive and EIA negative patients than HCV Qual (PCR) and should be part of our testing algorithm. PMID- 15125875 TI - Safety, tolerability, and plasma pharmacokinetics of high-strength formulations of enfuvirtide (T-20) in treatment-experienced HIV-1-infected patients. AB - BACKGROUND: Enfuvirtide, a HIV-1 membrane fusion inhibitor, is the first viral entry inhibitor approved for the treatment of HIV-1 infected patients in the USA. Parenteral administration of enfuvirtide in clinical trials has been safe and has resulted in significant decreases in plasma viral load, even in the setting of extensive previous treatment and multi-drug resistance to conventional antiretroviral (ARV) therapy. Previous formulations have required two injections administered twice-daily (BID). OBJECTIVES: The primary objectives of this study were to evaluate the safety, tolerability, and pharmacokinetics of two high strength 100 mg/ml formulations of enfuvirtide (carbonate [CO(3)] and tromethamine [TRIS] buffer) and of the current formulation (50 mg/ml CO(3) formulation) at doses of 90 mg (deliverable) BID and 67.5 mg (deliverable) BID in treatment-experienced patients. STUDY DESIGN: This was a phase II, multi-center, open-label, sequential cross-over pharmacokinetic, efficacy, and safety study. Study design included two treatment variables; dose (90 mg or 67.5 mg BID) and formulation (A: 50 mg/ml CO(3), B: 100 mg/ml CO(3) or C; 100 mg/ml TRIS). RESULTS: Forty-six treatment-experienced participants were sequentially enrolled into three treatment cohorts. All cohorts had similar safety profiles and only one patient discontinued due to an adverse event. Pharmacokinetic data indicated that the high-strength 100 mg/ml CO(3) formulation was bioequivalent to the 50 mg/ml CO(3) formulation whereas the TRIS formulation was not. At 48 weeks, 59.1%, 66.7% and 16.7% had <400 copies per milliliter HIV-1 RNA in the 90 MgCO(3), 67.5 MgCO(3) and 90 mg TRIS cohorts with median suppression of HIV-1 RNA of 2.97, 3.48, and 0.87 log(10)copies per milliliter, respectively. CONCLUSIONS: Based upon bioequivalence data and the convenience and similarity in safety and virological effect with the 50 mg/ml formulation, the 100 mg/ml CO(3) formulation was selected for use in clinical efficacy studies of enfuvirtide. PMID- 15125876 TI - Rapid detection and quantitation of hepatitis B virus DNA by real-time PCR using a new fluorescent (FRET) detection system. AB - BACKGROUND: The diagnosis of hepatitis B virus (HBV) has until recently been based on traditional serologic methods targeting viral antigens and antibodies to viral proteins. The development of molecular methods allowing for the quantitation of HBV DNA is proving clinically valuable for monitoring therapy and detecting early treatment failures. OBJECTIVES: Here we report a new real-time (LightCycler) quantitative PCR for the detection of HBV DNA based on sequence specific hybridisation probes (designed in-house), targeting the HBV surface antigen. STUDY DESIGN: The assay was evaluated using a 10-fold dilution series of standard HBV DNA [Eurohep standard reference 1, genotype A, HBsAg subtype adw with a unitage of 10(6) WHO. i.u./ml] and 89 clinical serum samples. The performance was measured against a quantified standard HBV DNA working reagent (NIBSC code 98/780) and the sensitivity compared with our conventional thermal block PCR. RESULTS AND CONCLUSION: Real-time PCR detected HBV DNA in 45% (40/89) and thermal-block PCR in 16% (14/75) of clinical samples. Results for 26 samples were below the detection limit of the thermal-block PCR but could be quantified by real-time (LightCycler) PCR. The LightCycler assay was at least 5 logs more sensitive than thermal-block PCR and could detect HBV in a linear range between 5 and 10(7) i.u. per reaction. The broad generic nature of the PCR primers coupled with the enhanced sensitivity and specificity of the fluorescent hybridisation probes makes this assay potentially valuable for both routine diagnostic and epidemiological work. PMID- 15125877 TI - Chronic excretion of a norovirus in a child with cartilage hair hypoplasia (CHH). AB - We have demonstrated the long-term excretion of a stable recombinant norovirus in a patient with cartilage hair hypoplasia (CHH), with a T cell immunodeficiency, following bone marrow transplantation (BMT). The patient excreted an ARG320/1999/US-like recombinant norovirus (rGII-3) for 156 days during a period of immune reconstitution. The child was symptomatic during the period of virus shedding. It is not known if the child acquired the recombinant strain or if recombination occurred in vivo. PMID- 15125878 TI - Detection of HIV-1 subtype G using Cobas Ampliscreen test. AB - A nucleic acid amplification test for detection of HIV-1 RNA was designed for screening pools of human plasma. This test achieves a similar level of sensitivity for group M subtypes, but few samples of subtype G and none of its CRFs had been tested, which are the most relevant in Portugal. We found that the test is effective in detecting HIV-1 subtypes and has an analytical sensitivity similar to B subtype. PMID- 15125879 TI - Is growth hormone stimulation testing in children still appropriate? AB - The diagnosis of growth hormone deficiency (GHD) historically has relied on measurement of growth hormone (GH) concentrations following stimulation, usually with a non-physiologic provocative agent. Despite the use of more specific GH assays, the peak concentration of GH below which a child is considered GH deficient has risen. We examine the pitfalls associated with GH stimulation tests, specifically, the lack of reliability and accuracy of these tests, and their inability to predict who will benefit from GH therapy. We recommend that GH stimulation tests no longer routinely be used for the diagnosis of GHD in children. PMID- 15125880 TI - Genetic strategies for elucidating insulin-like growth factor action in bone. AB - While it is universally accepted that IGFs serve important roles in bone acquisition and maintenance, the precise cellular targets and molecular mechanisms responsible for their actions have remained poorly understood. Recent advances in genetic techniques to manipulate the mouse genome have enabled the examination of individual components of the IGF system in vivo. Observations in these mouse models have provided new insights into the skeletal actions of IGF which have significant implications for understanding IGF action in human bone. PMID- 15125881 TI - Suppressors of cytokine signalling and regulation of growth hormone action. AB - The Suppressors of Cytokine Signalling (SOCS) are a family of proteins that are produced in response to signals from a diverse range of cytokines and growth factors and which act to attenuate cytokine signal transduction. Members of the SOCS family form a classical negative feedback loop with key actions involving inhibition of the Janus Kinase-Signal Transducers and Activators of Transcription (JAK-STAT) signalling cascade. Extensive analyses have implicated each of CIS, SOCS1, SOCS2 and SOCS3 in the regulation of Growth Hormone (GH) signal transduction. The expression of each of these SOCS proteins is induced in cells stimulated with GH and their over-expression in cell lines blocks aspects of GH signalling. In vivo studies with genetically modified mice have confirmed important physiological roles for SOCS proteins in regulation of GH action. In particular, mice lacking SOCS2 display gigantism accompanied by evidence of deregulated GH signalling. A precise understanding of the actions of SOCS proteins in GH signalling may offer new opportunities for therapeutic intervention in growth disorders and other conditions involving GH action. PMID- 15125882 TI - Three-years of growth hormone (GH) replacement therapy in GH-deficient adults: effects on quality of life, patient-reported outcomes and healthcare consumption. AB - OBJECTIVE: The objective was to investigate the effects of 3 years of growth hormone (GH) replacement therapy in GH deficient (GHD) patients in Sweden. DESIGN AND PATIENTS: An open label study in 237 adults with GHD (116 men and 121 women), consecutively enrolled in KIMS (Pfizer's international metabolic database) in Sweden. MEASUREMENTS: QoL and healthcare consumption were determined using questionnaires [QoL-assessment of GHD in Adults (QoL-AGHDA), the psychological general well-being (PGWB) index and the patient life situation form (PLSF)]. RESULTS: The mean starting dose of GH was 0.13 mg/day and the mean maintenance dose was 0.37 mg/day. The mean insulin-like growth factor I (IGF-I) SD score increased from -1.92 at baseline to 0.38 after 3 years. There was a sustained increase in QoL as measured by the QoL-AGHDA and PGWB questionnaires. The number of doctor visits and the number of days in hospital were reduced after 3 years of GH replacement. The number of days of sickleave decreased during the first 2 years of treatment, but returned towards baseline values after 3 years. Leisure time physical activity and satisfaction with physical activity increased. CONCLUSION: Three years of GH replacement therapy induced a sustained improvement in QoL. Healthcare consumption was reduced, although the reduction in the number of days of sickleave was not statistically significant after 3 years of treatment. PMID- 15125883 TI - Insulin-like growth factor binding protein-3 is a novel mediator of apoptosis in insulin-secreting cells. AB - Insulin-like growth factor binding protein-3 (IGFBP-3) is emerging as a critical regulator of cell survival. There has been no study which directly examined the potential role for this major growth factor in the programmed cell death (apoptosis) of insulin-secreting cells. To determine whether IGFBP-3 mediates apoptosis in insulin-secreting cells, we performed a rigorous series of experiments with the rat insulinoma (RIN) cell line m5F and the hamster insulin secreting tumor (HIT) T-15. Within 24 h exogenous IGFBP-3 induced significant DNA fragmentation in RIN and HIT cells, at doses ranging from 4.4 to 2000 ng/ml (P<0.05) without a classic dose-response relationship. DNA fragmentation induced by rhIGFBP-3 occurred in the presence of immunoglobulin to block the type 1 IGF receptor. As detected by flow cytometry for Annexin V exposure to the cell surface, rhIGFBP-3 treatment doubled the proportion of apoptotic HIT cells from 1.7 +/- 0.4% (serum-free control) to 3.4 +/- 0.2% (P<0.02), an effect completely reversed by co-treatment with 1000 ng/ml rhIGF-I. Immunofluorescent microscopy disclosed that pro-inflammatory Th1 cytokines increased intranuclear aggregation of endogenous IGFBP-3. Cytokine-induced DNA fragmentation was completely blocked by relatively brief pre-treatment with antisense IGFBP-3 phosphorothioate oligodeoxynucleotides. In conclusion, we have presented the first evidence that IGFBP-3 contributes to cytokine-mediated apoptosis in insulin-secreting cells. PMID- 15125884 TI - Pharmacodynamic evaluation of a PEGylated analogue of human growth hormone releasing factor in rats and pigs. AB - The aim of this study was to assess the in vivo efficacy of monoPEGylated GRF(1 29)NH(2) having one PEG(5000) chains attached to either lysine 12 or 21 as compared to the GRF(1-29)NH(2) in rats and pigs. This analogue termed GRF 1PEG(5000) was tested after a single intravenous administration in rats and after a single intravenous or subcutaneous injection in pigs. After 1 h administration, GH concentrations returned to values close to controls in the group of rats injected with GRF(1-29)NH(2). In animals injected with the same dose of GRF 1PEG(5000), the AUC values corresponding to the whole period 0.5-48 h and particularly to the 0.5-8 h period were higher than in the placebo or in the GRF(1-29)NH(2) groups. Interestingly, two additional peaks were observed at about 6 and 8 h following administration. An increase in the response of the endogenous GH peaks was also observed in pigs administered GRF-1PEG(5000) by intravenous route. When GRF-1PEG(5000) was administered subcutaneously to pigs, a significant increase, as compared to placebo and GRF(1-29)NH(2,) in both GH and IGF-I levels was observed. This new analogue might find therapeutic application in paediatric growth hormone deficiency or in aging. PMID- 15125885 TI - The insulin-like growth axis in patients with autoimmune thyrotoxicosis: effect of antithyroid drug treatment. AB - OBJECTIVE: Hyperthyroidism is associated with altered growth hormone (GH) secretion. Many patients with thyroid dysfunction experience several poorly described complications such as symptoms and signs also seen in patients with growth hormone deficiency (GHD). We have therefore prospectively evaluated a possible relationship between the thyroid function, body composition, leptin levels and insulin-like growth factor (IGF) related peptides in patients with Graves' disease. DESIGN, PATIENTS, AND MEASUREMENTS: In a prospective group of 24 fasting female patients with Graves' disease (mean age (CI 95%): 40 years (33 47)), we measured serum thyroxine, triiodothyronine, thyrotropine (TSH), TSH receptor antibodies, anti-thyroid peroxidase, leptin, body composition, body mass index (BMI) and IGF-related peptides at diagnosis and after 12 months of treatment with thiamazol (ATD). RESULTS: In thyrotoxic patients IGF-I plus IGF-II correlated positively with IGFBP-3 at baseline (r = 0.90, p < 0.1 x 10(16)) and after 12 months follow-up (r = 0.87, p < 0.1 x 10(-16)). In the thyrotoxic state total IGF-I, IGF-II, IGF binding protein 3 (IGFBP-3) and acid-labile subunit (ALS) but not free IGF-I decreased significantly from 223 microg/L (189-260) (mean (CI 95%), 877 microg/L (801-953), 4165 microg/L (3772-4577) and 22 mg/L (18 26)) to 198 microg/L (172-226), 788 microg/L (711-865), 3431 microg/L (3135-3741) and 19 mg/L (16-26) (p <0.006), respectively, after 12 months of ATD despite an increase in BMI from 22 (21-23) to 23 kg/m(2) (22-25) (p < 0.0004) but no significant changes in leptin. CONCLUSIONS: The complex IGF systems seemed intact in thyrotoxic patients but change in body composition and the regulation of leptin and insulin secretion during treatment of autoimmune thyroid disease influence IGF-related peptides leaving the patient in a state somewhat similar to partial GHD, but the mechanism behind these alterations remains unclear. PMID- 15125886 TI - IGF-I system responses during 12 weeks of resistance training in end-stage renal disease patients. AB - OBJECTIVE: To examine the hypothesis that 12 weeks of resistance training would alter circulating concentrations of IGF-I system components in end-stage renal disease (ESRD) patients. DESIGN: Ten ESRD patients underwent 12 weeks of resistance training after a 6 week control period and had morning fasted blood drawn on four occasions (weeks - 6, 0, 6, 12). Immunoassays were performed for serum total and free IGF-I, IGF binding proteins (IGFBPs) 2 and 3, and the acid labile subunit (ALS). Immunoaffinity depletion of ALS-based complexes allowed measurement of non-ternary (i.e., binary) IGF-I and IGFBP-3. RESULTS: Significant improvements in strength and functional performance were observed. All IGF-I measures were stable during the control period and no changes were observed for the first 6 weeks of resistance training. At week 12, total IGF-I (-15.4+/ 28.9%), ternary IGF-I (-16.4+/-36.7%), and the IGF-I/IGFBP-3 ratio had significantly (p < or = 0.05) declined from week 0 values. No changes were observed for free IGF-I, IGFBPs 2 and 3, or the acid labile subunit. The proportion of IGF-I in ternary ( approximately 76.3+/-6.8%), non-ternary ( approximately 22.5+/-6.6%), and free ( approximately 1.2+/-0.5%) forms remained constant throughout the training. CONCLUSIONS: 12 weeks of resistance training in ESRD patients induced a decline in total IGF-I, but did not alter the proportion of IGF-I circulating in free, ternary or non-ternary molecular complexes. The decline in IGF-I occurs in the presence of positive training adaptations on physical performance and we conclude that this response pattern appears to be reflective of favorable neuromuscular anabolic adaptations. PMID- 15125887 TI - Cardiac IGF-I manipulation by growth hormone following myocardial infarction. AB - Evidence of a role for growth hormone (GH) in cardiac structure and function has been derived from studies of patients suffering either GH excess or deficiency, both of which may lead to reduced life expectancy. The role of GH in the ischaemic heart, however, is less than clear. We therefore investigated the effect of 30 days GH treatment in sheep with myocardial infarction. GH treatment significantly increased circulating IGF-I levels (P<0.01), heart weight (P<0.01), and cardiomyocyte cross-sectional area (P<0.001). IGF-I mRNA in peri-infarct cardiac tissue also increased significantly (P<0.05). We conclude that post infarct GH treatment increases circulating and cardiac IGF-I levels, resulting in significant cardiomyocyte hypertrophy. This increase in cardiomyocyte size appears to correlate with local IGF-I expression rather than plasma IGF-I levels. PMID- 15125888 TI - G protein-coupled receptors: oligomerisation and association with accessory proteins. PMID- 15125889 TI - Selectivity in the oligomerisation of G protein-coupled receptors. AB - G protein-coupled receptors can exist as dimers and/or higher order oligomers. Such quaternary structure appears central to their plasma membrane delivery and, potentially, to function. Recent evidence that these receptors can form hetero- as well as homo-dimers/oligomers has significant implications for pharmacology and pathophysiology. Knowledge of the basis and selectivity of GPCR hetero dimerisation is thus vital. Current understanding of these areas is reviewed. PMID- 15125890 TI - Receptors for hypothalamic releasing hormones TRH and GnRH: oligomerization and interactions with intracellular proteins. AB - Studies of TRH and GnRH receptors have revealed much information about the roles of G-proteins and beta-arrestins, as well as receptor residues important for signaling, desensitization and internalization. However, the proteins involved are only just beginning to be identified and characterized. Additional complexity now exists with the observation that these receptors form oligomers in live cells. Indeed, hetero-oligomerization of TRH receptor subtypes 1 and 2 potentially alters interactions with intracellular regulatory proteins. Knowledge of proteins that interact with TRH or GnRH receptors will increase our understanding of receptor function and provide potential drug targets for a range of receptor-associated conditions. PMID- 15125891 TI - Beta-adrenergic receptors and their interacting proteins. AB - The three subtypes of beta-adrenergic receptor (beta AR) all interact with G proteins as a central aspect of their signaling. The various beta AR subtypes also associate differentially with a variety of other cytoplasmic and transmembrane proteins. These beta AR-interacting proteins play distinct roles in the regulation of receptor signaling and trafficking. The specificity of beta AR associations with various binding partners can help to explain key physiological differences between beta AR subtypes. Moreover, the differential tissue expression patterns of many of the beta AR-interacting proteins may contribute to tissue-specific regulation of beta AR function. PMID- 15125892 TI - Identification and functional roles of metabotropic glutamate receptor interacting proteins. AB - In the mammalian brain, a majority of excitatory synapses use glutamate as a neurotransmitter. Glutamate activates ligand-gated channels (ionotropic receptors) and G protein-coupled (metabotropic) receptors. During the past decade, a number of intracellular proteins have been described to interact with these receptors. These proteins not only scaffold the glutamate receptors at the pre- and post-synaptic membranes, but also regulate their subcellular targeting and intracellular signaling. Thus, identification of these proteins has been essential for further understanding the functions of glutamate receptors. Here we will focus on those proteins that interact with the subgroup of metabotropic glutamate (mGlu) receptors, and review the methods used for their identification, as well as their functional roles in neurons. PMID- 15125893 TI - The receptor activity modifying protein family of G protein coupled receptor accessory proteins. AB - Receptor diversity for the calcitonin peptide family is created by the interaction of two 7-transmembrane proteins--the calcitonin receptor (CTR) or the calcitonin receptor-like receptor (CL-R)--with the receptor activity modifying protein (RAMP) family. The discovery of heterodimeric complexes of these proteins heralded a new era in the study of G protein coupled receptors (GPCRs), whereby receptor phenotype is no longer governed by just the GPCR. In this article, recent advances in the study of RAMPs are discussed--from our current understanding of the molecular basis of RAMP-receptor interaction to a broader role for RAMPs outside the calcitonin receptor family. PMID- 15125894 TI - The role of G-protein coupled receptors and associated proteins in receptor tyrosine kinase signal transduction. AB - It is well established that stimulation of G-protein coupled receptors (GPCRs) can activate signalling from receptor tyrosine kinases by a process termed transactivation. Indeed, in recent years, it has become apparent that transactivation is a general phenomenon that has been demonstrated for many unrelated GPCRs and receptor tyrosine kinases. In this case the GPCR/G-protein participation is up-stream of the receptor tyrosine kinase. Substantial research has addressed these findings but meanwhile another mechanism of cross talk has been slowly emerging. For over a decade, a growing body of evidence has demonstrated that numerous growth factors use G-proteins and attendant signalling molecules such as beta-arrestins that participate down-stream of the receptor tyrosine kinase to signal to effectors, such as p42/p44 MAPK. This review highlights this novel mechanism of cross talk between receptor tyrosine kinases and GPCRs, which is distinct from growth factor receptor transactivation by GPCRs. PMID- 15125895 TI - Towards a perfect beta cell replacement. PMID- 15125896 TI - Navigating the pathway from embryonic stem cells to beta cells. AB - The compelling goal of using in vitro differentiation of stem cells to obtain replacement pancreatic beta cells that are clinically effective in treating diabetes has until now eluded researchers. This difficulty raises the question of whether more effective strategies are available. We propose that the native embryonic pathway leading to the definitive endoderm lineage, and continuing on to the endocrine pancreas, is the one most likely to succeed for the in vitro differentiation of embryonic stem cells. We question however whether gain-of function approaches involving genes necessary for beta cell development are destined to work effectively, and suggest alternative approaches to identifying conditions sufficient for in vitro beta cell differentiation. PMID- 15125897 TI - Making new beta cells from stem cells. AB - In 2000, Shapiro et al. provided compelling "proof of principle" data showing that the transplantation of human islets, purified from cadaveric material, could restore severely diabetic, Type 1 patients to insulin independence. This demonstration prompted renewed efforts to find an alternative and sustainable source of surrogate islet cells for cell therapy. Experiments involving adult ductal and liver "stem" cells, or embryonic stem cells, are prominent amongst these endeavors and are reviewed in this article. Whilst there are many published claims to success in converting ES cells into insulin secreting, glucose responsive cells, all require careful reinterpretation in the light of findings that cells can adsorb insulin present in growth media. It is likely that work with adult cells is less prone to this potential artifact and significant progress has been made in producing insulin-secreting cells. Assessment of in vivo function in the surrogate cells is most frequently made using cell transplantation into toxin-induced, diabetic mice, but this model is rarely used to maximal advantage. In many cases, it remains unclear whether reductions in the hyperglycemia result from insulin secretion from the transplanted cells or are due to recovery of endogenous islet function. In this latter context, experiments are reviewed where endogenous stimulation of recovery is engendered even by irradiated donor cells. PMID- 15125898 TI - Can we make surrogate beta-cells better than the original? AB - Insufficient pancreatic beta-cell mass is fundamental to the pathogenesis of both types 1 and 2 diabetes and constitutes the basis for the goal of beta-cell replacement therapy. Current methods for isolating islets from organ donor pancreases do not come close to supplying all in need, thus providing a compelling need to find new sources of insulin-producing cells. Possible sources include generation of cells from embryonic stem cells (ESC), adult stem/precursor cells, transdifferentiation of other cell types and xenodonors. Bioengineering can be used to improve secretory performance and strengthen cells to better withstand the challenges of transplantation. Strategies include protection against hypoxia, inflammation, and immune attack. PMID- 15125899 TI - Engineering a new beta-cell: a critical venture requiring special attention to constantly changing physiological needs. AB - As both type 1 and type 2 diabetes are characterized by reduced islet beta-cell mass and impaired insulin secretion, engineering new beta-cells for replacement therapy is appealing. For this to be successful, the intricate peptide processing and secretory machinery of the beta-cell must be duplicated. Further, the engineered beta-cell must be capable of modulating its function in response to physiological changes such as puberty, pregnancy and aging, and to more short term challenges such as infection, exercise and weight fluctuation. The new cell should have a low risk for recurrence of the primary disease and ensuring its survival should not be worse than diabetes itself. PMID- 15125901 TI - Hereditary breast and ovarian cancer syndrome: should we test adolescents? PMID- 15125902 TI - Chlamydia and gonorrhea screening in asymptomatic young women. AB - BACKGROUND: Adolescent and young adult minority women are at high risk for chlamydia (CT) and gonorrhea (NGC) cervical infections, which are significant causes of pelvic inflammatory disease, impaired fertility, ectopic pregnancy and chronic pain. The purpose of this article is to review among young women in the United States: (1) the epidemiology of CT and NGC cervical infection and their medical complications; (2) current public health recommendations to promote asymptomatic CT and NGC screening; (3) current screening practices and challenges of implementing public health recommendations; (4) testing and cost issues; and (5) future directions in promoting asymptomatic CT and NGC screening. METHODS: We conducted a MEDLINE search for articles published over the last two decades relating to CT and NGC screening in young women and then systematically reviewed all relevant articles. RESULTS: The data indicate that CT and NGC infection are geographically widespread in the U.S. and asymptomatic infection is highly prevalent among economically disadvantaged young females. Public health recommendations promoting CT and NGC screening in asymptomatic young women are directed to both health care providers and clients. However, strategies to promote screening efforts have been primarily directed toward health care providers; there are no published studies on client-initiated screening strategies. Challenges of implementing public health recommendations and future directions for CT and NGC screening are discussed. CONCLUSIONS: Young sexually active women continue to be at high risk for CT and NGC infection. The data indicate that implementation of health provider-based and client-initiated screening in private and public health care settings is a challenge. However, there is a great need to develop strategies to understand and promote client initiated screening. PMID- 15125903 TI - Vulvar venous malformations in an 11-year-old girl: a case report. AB - BACKGROUND: Unlike varicosities, which result from venous insufficiency, vascular malformations are developmental errors that do not regress. While these lesions are challenging to treat in most anatomic locations, genital venous malformations are particularly difficult problems for the gynecologist, urologist, or primary care physician who may identify them. The risk of surgical treatment has led to investigation of new therapeutic options for these vascular lesions. CASE: We describe an 11-year-old premenarchal female with bilateral, symptomatic vulvar venous malformations. These lesions were successfully treated with Doppler ultrasound-guided direct injection venography and ethanol sclerotherapy. CONCLUSION: Direct injection venography with ethanol sclerotherapy is an attractive diagnostic and therapeutic option for management of vulvar venous malformations. PMID- 15125904 TI - The use of the leukotriene receptor antagonist montelukast (Singulair) in the management of dysmenorrhea in adolescents. AB - PURPOSE: Previous studies have shown an increase in leukotrienes in the uterine tissue as well as in the menstrual flow of adult women with dysmenorrhea. An increase in leukotriene-E4, the major urinary leukotriene, was also reported in adolescent girls with dysmenorrhea, further suggesting a possible involvement of these potent vasoconstrictors and inflammatory mediators in generating dysmenorrhea symptoms. In the present study we examined whether blocking leukotrienes might alleviate symptoms of dysmenorrhea in adolescents. METHODS: Twenty-five adolescents (age 16 +/- 1 years, 4 +/- 1 years post menarche, body mass index 23 +/- 1) with dysmenorrhea participated in a randomized, double blind, crossover study. Thirteen girls received one tablet of montelukast (Singulair, Merck, West Point, PA) 10 mg daily starting on day 21 of the cycle until the last day of the menstrual period for two menstrual cycles, followed by one tablet of placebo (Merck, West Point, PA) daily starting on day 21 of the cycle until the last day of the menstrual period for two additional menstrual cycles. The other 12 girls had a reverse schedule starting with placebo. Participants were instructed to use one or two 200-mg tablets of ibuprofen every 6 h in the event of continuing menstrual symptoms. The Cox Menstrual Symptom Scale was used to assess response to treatment. An intent-to-treat approach was used for data analysis. RESULTS: Twenty-two girls completed the study. Two girls were noncompliant with the study protocol, and one was withdrawn because of Helicobacter pylori infection. Compared with Cox menstrual score (mean +/- SE) before study (46 +/- 6), there was no significant change in menstrual symptoms during treatment with placebo (Cox score 42 +/- 7) or during treatment with montelukast (Cox score 39 +/- 7), and there was no significant difference between montelukast and placebo treatments as well. Likewise, there was no significant difference between the amount of ibuprofen tablets consumed during the menstrual periods before study (4 +/- 1), while on placebo (3 +/- 1), and while on montelukast (4 +/- 1). CONCLUSIONS: This study does not support the use of montelukast, in the current FDA-approved dose (for asthma) and commencing immediately before the menstrual period, for treatment of dysmenorrhea. It remains to be determined in further studies whether a higher dose or a prolonged daily use of montelukast may alleviate symptoms of dysmenorrhea in adolescents. PMID- 15125905 TI - Immature ovarian teratoma in an adolescent: a case report and review of the literature. AB - BACKGROUND: Benign cystic teratomas are relatively common tumors in reproductive age women, but can occur at any age. While the incidence of malignant elements in a teratoma is low (approximately 1-2%), the survival of patients with immature teratoma is poor. Definitive diagnosis is mandatory. CASE: We describe a case of a 13-year-old African American female, gravida 0, presenting with a large pelvic mass, determined to be a benign cystic teratoma by intra-operative frozen section. However, due to the size of the tumor and the preponderance of neural elements we performed a full surgical staging procedure (excluding hysterectomy and complete removal of adnexa). The final pathology report revealed foci of immature neural tissue, with a final diagnosis of an immature cystic teratoma Stage Ia. CONCLUSION: Foci of immature neural elements can be readily missed on frozen section, especially with a large tumor. Full surgical staging at the time of initial laparotomy is justified when encountering an apparently mature cystic teratoma with a preponderance of neural elements on frozen section. PMID- 15125906 TI - Medical evaluation of suspected child sexual abuse. PMID- 15125907 TI - Toolkit for adolescents. PMID- 15125908 TI - Tips for clinicians. Sexually transmitted infections in adolescents: a treatment update. PMID- 15125909 TI - Perspectives on pediatric and adolescent gynecology from the allied health care professional. Teens, tattoos and body piercing. PMID- 15125910 TI - Sensory cueing effects on maximal speed gait initiation in persons with Parkinson's disease and healthy elders. AB - Researchers have suggested that sensory cues can improve gait initiation in persons with Parkinson's disease (PD); however, there is little research that documents the effects of sensory cues on gait initiation. The purpose of this study was to examine the effects of auditory and cutaneous sensory cues on maximal speed gait initiation in person's with PD and healthy elderly. Seven persons with PD of moderate severity (mean age=69 years) and seven age, gender, and height matched healthy elders participated. Temporal, kinematic and center of pressure (COP) data were recorded as participants performed eight trials within four randomly ordered conditions (no cue (NC), a single auditory cue (SA), repetitive auditory cues (RA), and repetitive cutaneous cues (RC)). In each condition, participants were instructed to perform each gait initiation trial at their maximal speed. In all conditions, person's with PD reacted more slowly and moved less far than did the matched elders. Relative to conditions with NCs, sensory cueing resulted in decreased double limb support (DLS), and increased COP displacement and velocity in both groups. However, in both groups, displacements and velocities of the swing limb and sacrum during the sensory-cued conditions were less than those during the NC condition. These results suggest that when movement speed is a primary goal, sensory cues may interfere with swing limb and body movement outcomes during the gait initiation task in both person's with PD and healthy elders. PMID- 15125911 TI - Obstacle negotiation kinematics: age-dependent effects of postural threat. AB - This study investigated whether obstacle negotiation kinematics among younger (YA) and older adults (OA) are influenced by postural threat. Obstacle negotiation kinematics among YA and OA were examined under four conditions of postural threat. Seventeen older and 15 YA negotiated a fixed virtual obstacle while walking at a self-determined velocity along a 7.2 m walkway. Postural threat was manipulated by varying the width (0.60 vs. 0.15 m) and height (0.00 vs. 0.60 m) of the walkway. Increasing postural threat resulted in elevated levels of physiological arousal and altered crossing kinematics for all subjects. Specifically, crossing step length, lead limb velocity, trail limb velocity, and whole body center of mass (COM) velocity decreased and lead limb crossing height increased in the condition of greatest postural threat compared with the condition of least postural threat. Although both YA and OA altered obstacle negotiation kinematics under conditions of postural threat, the changes observed among older adults were considerably different from those of YA. In particular, OA demonstrated more marked reductions in the crossing velocity of the lead limb, trail limb, and COM than YA between the condition of least postural threat and the condition of greatest postural threat. The results of this study reveal that postural threat influences negotiation kinematics among YA and OA and illustrate that age-dependent differences exist for obstacle negotiation kinematics when postural threat increases. These findings may imply a beneficial effect of fear of falling on reducing fall probability among healthy OA. PMID- 15125912 TI - Evaluation of early walking patterns from plantar pressure distribution measurements. First year results of 42 children. AB - This study evaluated developmental changes of foot and gait during the first year of independent walking. In a longitudinal design, plantar pressure distribution patterns were measured with a capacitive platform every 3 months in a group of 42 normal children. The first significant changes were already found after a few weeks of independent walking. The development of the longitudinal arch correlated with significantly reduced midfoot loading parameters even though the changes showed a wide interindividual variation. This study illustrates that a child's foot goes through significant changes in shape and loading characteristics once the child starts to stand and walk. Some children showed a fairly mature appearance of the plantar pressure pattern after 1 year while others retained a more immature loading pattern. PMID- 15125913 TI - Gait termination in young and older adults: effects of stopping stimulus probability and stimulus delay. AB - Sixteen young (25 +/- 2.6 years) and 16 older individuals (69 +/- 4.4 years) walked normally then terminated walking rapidly. A visual stopping stimulus was presented 10 ms following ground contact (short delay) and in another condition, at 450 ms prior to toe-off (long delay). Stimulus probability was either high (80% of trials) or low (10%). The younger group stopped faster (463 vs. 574 ms) despite also walking faster (1.29 vs. 1.17 m s(-1)). Longer delay decreased one step responses but older participants used significantly more (slower) two-step stopping, which increased stopping time and distance. The additional step may have been pre-planned to maintain medial-lateral stability. PMID- 15125914 TI - Is a "loss of balance" a control error signal anomaly? Evidence for three-sigma failure detection in young adults. AB - Given that a physical definition for a loss of balance (LOB) is lacking, the hypothesis was tested that a LOB is actually a loss of effective control, as evidenced by a control error signal anomaly (CEA). A model-reference adaptive controller and failure-detection algorithm were used to represent central nervous system decision-making based on input and output signals obtained during a challenging whole-body planar balancing task. Control error was defined as the residual generated when the actual system output is compared with the predicted output of the simple first-order polynomial system model. A CEA was hypothesized to occur when the model-generated control error signal exceeded three standard deviations (3sigma) beyond the mean calculated across a 2-s trailing window. The primary hypothesis tested was that a CEA is indeed observable in 20 healthy young adults (ten women) performing the following experiment. Seated subjects were asked to balance a high-backed chair for as long as possible over its rear legs. Each subject performed ten trials. The ground reaction force under the dominant foot, which constituted the sole input to the system, was measured using a two axis load cell. Angular acceleration of the chair represented the one degree-of freedom system output. The results showed that the 3sigma algorithm detected a CEA in 94% of 197 trials. A secondary hypothesis was supported in that a CEA was followed in 93% of the trials by an observable compensatory response, occurring at least 100 ms later, and an average of 479 ms, later. Longer reaction times were associated with low velocities at CEA, and vice versa. It is noteworthy that this method of detecting CEA does not rely on an external positional or angular reference, or knowledge of the location of the system's center of mass. PMID- 15125915 TI - High-arched runners exhibit increased leg stiffness compared to low-arched runners. AB - Leg stiffness between high-arched (HA) and low-arched (LA) runners was compared. It was hypothesized that high-arched runners would exhibit increased leg stiffness, increased sagittal plane support moment, greater vertical loading rates, decreased knee flexion excursion and increased activation of the knee extensor musculature. Twenty high-arched and 20 low-arched subjects were included in this study. Leg stiffness, knee stiffness, vertical loading rate and lower extremity support moment were compared between groups. Electromyographic data were collected in an attempt to explain differences in leg stiffness between groups. High-arched subjects were found to have increased leg stiffness and vertical loading rate compared to low-arched runners. Support moment at the impact peak of the vertical ground reaction force was related to leg stiffness across all subjects. High-arched subjects demonstrated decreased knee flexion excursion during stance. Finally, high-arched subjects exhibited a significantly earlier onset of the vastus lateralis (VL) than the low-arched runners. Differences exist in leg stiffness and vertical loading rate between runners with different foot types. Differences in lower extremity kinetics in individuals with different foot types may have implications for new treatment strategies or preventative measures. PMID- 15125916 TI - Speed related changes in muscle activity from normal to very slow walking speeds. AB - The study of neuromuscular activity at very slow walking speeds may lead to a better understanding of the mechanisms underlying speed regulation during walking, and may aid the interpretation of gait data in patients who walk slowly. Extreme reductions in walking speed will cause changes in locomotor task demands that may eventually result in modifications of the patterning of muscle activity that underlies walking. The aim of the present study was to investigate patterns of lower limb muscle activity during very slow walking (< 0.28 m s(-1)), and to study the neuromuscular gain functions that reflect the phase dependent effects of walking speed on electromyographic (EMG) amplitude. Nine healthy young adults walked at seven different walking speeds (1.39, 0.83, 0.28, 0.22, 0.17, 0.11, and 0.06 m s(-1)) while EMG was recorded from eight lower extremity muscles. Results showed that the phasing of muscle activity remained relatively stable over walking speeds despite substantial changes in its amplitude. However, between 1.39 and 0.28 m s(-1), epochs of Rectus femoris, Biceps femoris and Tibialis anterior activities were found that were typical for specific speed ranges. When walking speed decreased further to almost standing still (0.06 m s(-1)), negative gain values were found in Peroneus longus during midstance and Rectus femoris in late swing, indicating the emergence of new bursts of activity with decreasing walking speed. It is proposed that these extra activities may be attributed to increased demands on postural stability, and the altered dynamics of the swinging limb at very slow speeds. PMID- 15125917 TI - Characteristics of stepping over an obstacle in community dwelling older adults under dual-task conditions. AB - Previous research suggests that older adults may have difficulty attending to simultaneous tasks. This study was conducted to determine how concurrent performance of a secondary cognitive task influences walking and stepping over an obstacle in community dwelling older adults. Twenty-one men and women with a mean age of 73.4 years (S.D.=5.3) participated in the study. Subjects performed a gait task both alone (single-task condition) and in combination with a cognitive task that involved reciting numbers (dual-task condition). In the gait task, each subject walked at his/her fastest speed along a 10-m walkway and stepped over an obstacle designed to simulate a door threshold. Paired t-tests were used to compare gait parameters (10 m gait speed, gait speed during obstacle approach and negotiation, medial-lateral center of pressure excursion and velocity during obstacle negotiation, foot clearance over the obstacle, step length and foot position relative to the obstacle) and cognitive task performance under single and dual-task conditions. Toe-obstacle distance was greater and obstacle-heel distance was reduced under dual-task conditions. Performance of the remaining gait parameters did not change with the addition of a secondary cognitive task. Cognitive task performance decreased under dual-task conditions. These community dwelling older adults demonstrated minimal or no change in measured gait parameters during simultaneous performance of a cognitive task. The observed decrement in cognitive task performance suggests that subjects may have placed a higher priority on gait performance. PMID- 15125918 TI - Test-retest reliability of trunk accelerometric gait analysis. AB - The purpose of this study was to determine the test-retest reliability of a trunk accelerometric gait analysis in healthy subjects. Accelerations were measured during walking using a triaxial accelerometer mounted on the lumbar spine of the subjects. Six men and 14 women (mean age 35.2; range 18-57) underwent the same protocol on 2 consecutive days. The raw acceleration signals from six self selected walking speeds were transformed into a horizontal-vertical coordinate system to remove unwanted variability caused by gravity. Acceleration root mean square values, cadences, step and stride lengths were then computed and interpolated using quadratic curve fits and point estimates were calculated at a standardised walking speed of 1.35 m/s. Relative reliability was determined using two models of intraclass correlation coefficients (ICC(1,1) and ICC(3,1)) to assess any systematic shifts and absolute reliability was determined using measurement error (ME). The results of the study showed high ICC values (0.77 0.96) and ME values of 0.007-0.01 g for mean acceleration; 0.009 m for step lengths; 0.022 m for stride length and 1.644 step/min for cadences. In conclusion, the method was found to be reliable and may have a definite potential in clinical gait analysis. PMID- 15125919 TI - Distal hamstring lengthening in ambulatory children with cerebral palsy: primary versus revision procedures. AB - To document the benefits and limitations of distal hamstring lengthening (HL), 61 children (105 limbs) with cerebral palsy treated by distal HL with complete preoperative and postoperative evaluations were reviewed. There was significant improvement in popliteal angle, fixed knee flexion contracture, knee angle at foot contact (FC), and mid-stance knee extension after HL. On the other hand, the hip power generation peak decreased, and the anterior pelvic tilt increased. For the repeated HL (22 limbs), the fixed knee flexion contracture and knee flexion at FC improved. In a group of ten patients (17 limbs) with further postoperative follow up evaluations, the only significant clinical finding related to hamstring function between the first postoperative and the follow up evaluations with no intervening surgery was an increase in the popliteal angle of 20 degrees. Because the popliteal angle may increase over time after HL with no other directly related gait changes, the indications for repeated HL should include fixed knee flexion contracture and increased knee flexion at FC. The increase in the popliteal angle alone should not be considered an indication for repeated HL. PMID- 15125920 TI - Novel 3,5-diaryl pyrazolines as human acyl-CoA:cholesterol acyltransferase inhibitors. AB - A series of pyrazoline derivatives were prepared for evaluating their acyl CoA:cholesterol acyltransferase activities. 3-(3,5-Di-tert-butyl-4-hydroxyphenyl) 5-(multi-substituted 4-hydroxyphenyl)-2-pyrazolines 4a-i were shown in vitro inhibitory activity on hACAT-1 and -2. PMID- 15125921 TI - Novel 3,5-diaryl pyrazolines and pyrazole as low-density lipoprotein (LDL) oxidation inhibitor. AB - Compounds 4a-j and 5 were synthesized by cyclocondensation of 3a-j and hydrazine and showed significant LDL-antioxidant activities in the TBARS assay, the lag time of conjugated diene production, the relative electrophoretic mobility (REM) of ox-LDL, the apoB-100 fragmentation, and the macrophage-mediated LDL oxidation. Among compounds 4a-j and 5, 4a was found to be the most active compound as an inhibitor of LDL oxidation and 4a (IC50 = 0.1 microM) was 6-fold more potent than probucol (IC50 = 0.6 microM) in the TBARS assay. PMID- 15125922 TI - Synthesis and characterization of bifunctional probes for the specific labeling of fusion proteins. AB - Labeling proteins with synthetic probes is important for studying and characterizing protein function. We have recently introduced a general method for the specific in vivo and in vitro labeling of fusion proteins that is based on the reaction of O6-alkylguanine-DNA alkyltransferase (AGT) with O6-benzylguanine derivatives. Here we report two complementary routes for the synthesis of O6 benzylguanine derivatives, which allow for the labeling of AGT fusion proteins with bifunctional synthetic probes and demonstrate the specific labeling of AGT fusion proteins with these probes. These molecules should become useful tools for various applications in functional proteomics. PMID- 15125923 TI - Tetrahydroisoquinolines as subtype selective estrogen agonists/antagonists. AB - Two series of 6-hydroxy and 7-hydroxy tetrahydroisoquinolines were prepared. Evaluating a range of C-1, C-4, and N-substituents led to the discovery of ER alpha and ER beta selective analogs. PMID- 15125924 TI - Initial structure-activity relationships of lysophosphatidic acid receptor antagonists: discovery of a high-affinity LPA1/LPA3 receptor antagonist. AB - A recently reported dual LPA1/LPA3 receptor antagonist (VPC12249, 1) has been modified herein so as to optimize potency and selectivity at LPA receptors. Compounds containing variation in the acyl lipid chain and linker region have been synthesized and screened for activity at individual LPA receptors. LPA1 selective (14b) and LPA3-selective (10g,m) compounds of modest potency have been discovered. Additionally, 2-pyridyl derivative 10t exhibits a Ki value of 18 nM at the LPA1 receptor and is significantly more potent than 1 at the LPA3 receptor. This paper describes the synthetic methods, biological evaluation, and structure-activity relationships (SARs) of LPA receptor antagonists. PMID- 15125925 TI - Estrogen receptor ligands. Part 4: The SAR of the syn-dihydrobenzoxathiin SERAMs. AB - A series of estrogen receptor ligands based on a dihydrobenzoxathiin scaffold is described and evaluated for estrogen/anti-estrogen activity in both in vitro and in vivo models. The most active analogue, 22, was found to be 40-fold ERalpha selective in a competitive binding assay, and 22 demonstrated very potent in vivo antagonism of estradiol driven proliferation in an immature rat uterine weight gain assay. PMID- 15125926 TI - Potent nonpeptide vasopressin receptor antagonists based on oxazino- and thiazinobenzodiazepine templates. AB - Vasopressin receptor antagonists can elicit ion-sparing diuretic effects (i.e., aquaresis) in vivo by blunting the action of the circulating hypophyseal hormone arginine vasopressin. We have identified two new series of basic tricyclic benzodiazepines, represented by general structure 1, which contain compounds that bind with high affinity to human V2 receptors. For example, (S)-(+)-8 and 5 are potent and selective V2 receptor antagonists with pronounced aquaretic activity in rats on oral administration. PMID- 15125927 TI - Studies on aromatic compounds: inhibition of calpain I by biphenyl derivatives and peptide-biphenyl hybrids. AB - With the objective to understand structural features responsible for the biological activity, novel nonelectrophilic biphenyl derivatives and peptide biphenyl hybrids have been synthesized and evaluated as calpain I inhibitors. The preliminary results indicate that the presence of additional aromatic rings (besides the biphenyl system) makes these compounds potent calpain inhibitors with IC50 values in the nanomolar range. PMID- 15125928 TI - Design and synthesis of benzofused heterocyclic RXR modulators. AB - Benzofused heterocyclic analogs of the RXR selective modulator 1 (LG101506) were synthesized, and tested for their ability to bind RXRalpha and activate RXR homo and heterodimers. Potency and efficacy were observed to be dependent upon the choice of heterocycle as well as the sidechain employed. PMID- 15125929 TI - Synthesis and structure-activity relationship of 2-(aminoalkyl)-3,3a,8,12b tetrahydro-2H-dibenzocyclohepta[1,2-b]furan derivatives: a novel series of 5 HT(2A/2C) receptor antagonists. AB - Following the program started at Johnson & Johnson Pharmaceutical Research & Development searching for 5-HT(2A/2C) antagonists we now report on the synthesis of a series of substituted 2-(aminomethyl)-3,3a,8,12b-tetrahydro-2H dibenzocyclohepta[1,2-b]furan derivatives. The 5-HT2A, 5-HT2C and H1 receptor affinities of the described compounds are reported. The mCCP antagonistic activity of a set of selected molecules is also reported. PMID- 15125930 TI - Synthesis of new fluoroquinolones and evaluation of their in vitro activity on Toxoplasma gondii and Plasmodium spp. AB - The synthesis of four new computer-designed fluoroquinolones which have been predicted by QSAR analysis to be active against the protozoa Toxoplasma gondii is described. These compounds are inhibitory in vitro for T. gondii. One of these compounds has a remarkably high activity comparable to that of trovafloxacin. It combines the basic cyclopropyl-quinoline structure of gatifloxacin or moxifloxacin with the C-7 6-amino-3-azabicyclo[3.1.0]hexyl side chain of trovafloxacin. The four compounds are also inhibitory for blood stages of Plasmodium falciparum though at high concentration. These results confirm the potential of quinolones as anti-T. gondii and antimalarial drugs but also show that the QSAR models for T. gondii cannot be reliably extended for screening antimalarial activity. PMID- 15125931 TI - The significant effect of the carbohydrate structures on the DNA photocleavage of the quinoxaline-carbohydrate hybrids. AB - The novel DNA interactive quinoxaline-carbohydrate hybrids possessing disaccharides as the carbohydrate moieties were designed and synthesized, and their DNA photocleaving abilities were evaluated in order to examine the effect of the disaccharide structures. The configurations of the glycosidic bonds in the disaccharide strongly affected the DNA photocleaving ability, and two beta glycosidic bond linkages were very effective for the DNA photocleavage. Furthermore, the quinoxaline-disaccharide hybrids exhibited selective cytotoxicity against cancer cells with photoirradiation. PMID- 15125932 TI - Synthesis of OSW-1 analogs with modified side chains and their antitumor activities. AB - Four analogs of OSW-1 (1-4) with modified side chains on the steroidal skeleton were synthesized following modification of our previous route for the total synthesis of OSW-1. Testing of the analogs against growth of tumor cells demonstrated that the 22-one function and the full length of the side chain of OSW-1 were not required for the antitumor action of OSW-1. PMID- 15125933 TI - In vivo MR detection of vascular endothelial injury using a new class of MRI contrast agent. AB - A new class of dye-based MRI contrast agents, EB-DTPA-Gd, was designed and synthesized. The contrast agent was found to accumulate at the site of endothelial injury when the reagent was applied to isolated porcine blood vessels or in an ex vivo experiment using rat. In vivo MR detection of vascular endothelial injury was also successful in rat with its common carotid artery injured by balloon treatment. These results indicate that EB-DTPA-Gd is potentially useful for the diagnosis of vascular diseases. PMID- 15125934 TI - 2,4-Bis(octadecanoylamino)benzenesulfonic acid sodium salt as a novel scavenger receptor inhibitor with low molecular weight. AB - In order to investigate the effect of the fixation of the orientations of the two long chains, three types of novel derivatives of scavenger receptor inhibitor 1 were synthesized, and their biological activities were evaluated. Among the novel derivatives, 2,4-bis(octadecanoylamino)benzenesulfonic acid sodium salt (4d) showed the most potent inhibitory activity against the incorporation of 1,1' dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate-labeled acetyl-LDL (DiI-acetyl-LDL) into macrophages. 2,5-Bis(octadecanoylamino)benzenesulfonic acid sodium salt (4c), a regioisomer of 4d, did not exhibit as potent an inhibitory activity as 4d, meaning that the substitution pattern of two long chains on the benzene ring must be important. Compound 4d exhibited 10 times more potent inhibitory activity against the binding of 125I-labeled acetyl-LDL to the surface of macrophages than compound 1. PMID- 15125935 TI - 1- and 2-substituted naphthalenes: a new class of potential hypotensive agents. AB - A series of 2-substituted aminomethyloxy naphthalenes 1 and 4-(1-naphthoxy-2 substituted aminomethyl)-butanoic acids 2 were synthesized by Mannich reaction of 4-(2-naphthoxy)-butanoic acid 3 and 4-(1-naphthoxy)-butanoic acid 4 with appropriate secondary amines and para-formaldehyde. The newly synthesized compounds were tested for their hypotensive activity at 5 mg/kg i.v. dose in cats. The results indicated that the analogue 2-(N4-phenyl-N1-piperazino) methyloxy naphthalene 1d (> N = N4-phenyl-N1-piperazino) was the most active analogue when its hypotensive activity was compared to the reference compound propranolol. PMID- 15125936 TI - Novel factor Xa inhibitors based on a benzoic acid scaffold and incorporating a neutral P1 ligand. AB - A series of novel, highly potent, achiral factor Xa inhibitors based on a benzoic acid scaffold and containing a chlorophenethyl moiety directed towards the protease S1 pocket is described. A number of structural features, such as the requirements of the P1, P4 and ester-binding pocket ligands were explored with respect to inhibition of factor Xa. Compound 46 was found to be the most potent compound in a series of antithrombotic secondary assays. PMID- 15125937 TI - Direct superoxide anion scavenging by a highly water-dispersible carotenoid phospholipid evaluated by electron paramagnetic resonance (EPR) spectroscopy. AB - Synthetic carotenoid analogs, with increased utility for biological applications, are sparingly reported in the literature. Synthetic modification, which may increase the water solubility and/or water dispersibility of lipophilic carotenoids, allows their use in aqueous environments as potent antioxidants against potentially deleterious reactive oxygen species (ROS) that can be generated in vivo. Superoxide anion, produced by activated human neutrophils, can be a source of additional harmful ROS and nonradical species such as singlet oxygen in vivo. In the current study, direct scavenging of superoxide anion by a well-characterized C30 carotenoid phospholipid mixture was evaluated in a standard in vitro isolated human neutrophil assay by electron paramagnetic resonance (EPR) spectroscopy, employing the spin-trap DEPMPO. The carotenoid phospholipid was tested in aqueous formulation (aqueous dispersibility >60 mg/mL), in which supramolecular assembly takes place, as well as in ethanolic formulation as a monomeric solution of the carotenoid phospholipids. The carotenoid phospholipid (a highly unsaturated zwitterionic surfactant) was compared with a previously characterized rigid, long-chain, highly unsaturated dianionic bolaamphiphile, which contains an additional three conjugated double bonds in its extended conjugated system. As previously reported, direct scavenging by the carotenoid phospholipid derivatives in monomeric ethanolic formulation was superior at each tested concentration to aqueous, aggregated formulations of the compounds. Additionally, the percent inhibition of superoxide signal was related to the apparent or effective length of the conjugated chromophore, consistent with previous reports of radical inhibition and singlet oxygen quenching by polyene carotenoids of differing length. PMID- 15125938 TI - Synthesis of novel fluorescent-labelled dinucleoside polyphosphates. AB - A novel tandem synthetic-biosynthetic procedure is described for the synthesis of four new fluorescent dinucleoside polyphosphates: mant-Ap4A, mant-AppCH2ppA, TNP Ap4A and TNP-AppCH2ppA. These compounds are expected to supplement the existing etheno (epsilon) and 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) labelled derivatives, being the fluorescent probes of choice to investigate polyphosphate/enzyme binding behaviour. PMID- 15125939 TI - Hit-to-lead studies: the discovery of potent, orally active, thiophenecarboxamide IKK-2 inhibitors. AB - A hit-to-lead optimisation programme was carried out on the thiophenecarboxamide high throughput screening hits 1 and 2 resulting in the discovery of the potent and orally bioavailable IKK-2 inhibitor 22. PMID- 15125940 TI - Pharmacophore-based search, synthesis, and biological evaluation of anthranilic amides as novel blockers of the Kv1.5 channel. AB - The search for novel, potent Kv1.5 blockers based on an anthranilic amide scaffold employing a pharmacophore-based virtual screening approach is described. The synthesis and structure-activity relationships (SAR) with respect to inhibition of the Kv1.5 channel are discussed. The most potent compounds display sub-micromolar inhibition of Kv1.5 and no significant effect on the HERG channel. In addition, good oral bioavailability is demonstrated for compound 3i in rats. PMID- 15125941 TI - Stability studies of C-4',6' acetal benzylmaltosides synthesized as inhibitors of smooth muscle cell proliferation. AB - In our investigations to synthesize inhibitors of smooth muscle cell (SMC) proliferation, compound 6a displayed submicromolar activity in in vitro antiproliferative assays and reduced intimal thickening using a rat balloon angioplasty model via i.v. administration. In order to identify analogs that could be administered orally, the chemical instability of the C-4',6' acetal of compound 6a was addressed. Several novel variants with increased acid stability and comparable in vitro activity were prepared. PMID- 15125942 TI - A facile route to dynamic glycopeptide libraries based on disulfide-linked sugar peptide coupling. AB - We report here that disulfide-linked dynamic glycopeptide libraries can be constructed from 1-thiosugar and cysteine-rich oligopeptide building blocks upon gentle air oxidation of a slightly basic (pH 7.8) aqueous solution thereof. A mixture of 1-thiogalactose and two oligopeptides H2N-CysGlyCysGly-CO2H and H2N GlyCycCysGlyGly-CO2H, for example, affords a poorly HPLC-resolved disulfide library composed of various sugar-peptide conjugates and cyclic peptides, at least 10 of which can be identified by ESI mass spectrometry. The building components of disulfide members are exchangeable with each other in the presence of dithiothreitol as an initiator to allow dynamic equilibration. A preliminary SPR examination shows that the thiogalactose-derived library indeed contains active divalent galactoside species capable of cross-linking peanut lectin molecules. PMID- 15125943 TI - The oxime bond formation as an efficient chemical tool for the preparation of 3',5'-bifunctionalised oligodeoxyribonucleotides. AB - The simultaneous conjugation of peptides or carbohydrates at the 3'- and 5'-end of oligodeoxyribonucleotides was achieved very efficiently through chemoselective oxime bond formation. The method employs bifunctionalised oligonucleotides in single step without the need of protection strategy, under mild acidic conditions. The conjugates were obtained in high yields by reacting an oxyamine containing reporter groups (peptide, mono- and disaccharide) with an oligonucleotide carrying an aldehyde at each extremity. PMID- 15125944 TI - Solid-phase synthesis of kojic acid-tripeptides and their tyrosinase inhibitory activity, storage stability, and toxicity. AB - A small library of kojic acid-tripeptides (Ko-X1X2X3) was prepared by solid-phase parallel synthesis and assayed to evaluate their tyrosinase inhibitory activity. Most of the kojic acid-tripeptides showed better activities than kojic acid. Kojic acid-FWY was the best compound, and it exhibited 100-fold tyrosinase inhibitory activity compared with kojic acid. In addition, their storage stabilities were approximately 15 times higher and their toxicity was lower than that of kojic acid. PMID- 15125945 TI - PDE2 inhibition by the PI3 kinase inhibitor LY294002 and analogues. AB - Synthetic 2-morpholinochromones, including the known PI3-kinase inhibitor LY294002, have been evaluated in vitro as inhibitors of isolated human platelet phosphodiesterases. Inhibition of the cAMP-phosphodiesterases, PDE2 and PDE3 by LY294002 is reported for the first time. Preliminary screening across a range of 2-morpholinochromones has revealed structural features for optimised PDE2 inhibition. PMID- 15125946 TI - Silanediol peptidomimetics. Evaluation of four diastereomeric ACE inhibitors. AB - Four diastereomers of a Phe-Ala peptide mimic incorporating a central silanediol group have been individually prepared and tested as inhibitors of angiotensin converting enzyme (ACE). Three of the silanediols exhibit levels of inhibition that are similar to those of corresponding ketones reported by Almquist. For the fourth diastereomer, with both stereogenic carbons inverted relative to the most active isomer, the ketone gives the least enzyme inhibition whereas the silanediol shows a surprisingly low IC50 value. PMID- 15125947 TI - Design, synthesis and structure-activity relationship studies of novel indazole analogues as DNA gyrase inhibitors with Gram-positive antibacterial activity. AB - In this study, we report the design, synthesis and structure-activity relationships of novel indazole derivatives as DNA gyrase inhibitors with Gram positive antibacterial activity. Our results show that selected compounds from this series exhibit potent antibacterial activity against Gram-positive bacteria including multi-drug resistant strains that is methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VRE). PMID- 15125948 TI - Potent DNA gyrase inhibitors; novel 5-vinylpyrazole analogues with Gram-positive antibacterial activity. AB - In this study, we designed and synthesized novel 5-vinylpyrazole analogues by decreasing the lipophilicity of the parent compounds 1a,b; 3-[(3 methoxycarbonyl)cyclohexylaminomethyl]indazoles while keeping the van der Waals interaction with the lipophilic area of DNA gyrase B. The selected compound 8bb exhibited good antibacterial activity against staphylococci and enterococci, including multi-drug resistant strains. PMID- 15125949 TI - Chemo- and stereoselective synthesis of benzocycloheptene and 1-benzoxepin derivatives as alpha-sympathomimetic and anorexigenic agents. AB - The synthesis and pharmacological evaluation of cis- and trans-6-amino-6,7,8,9 tetrahydro-5H-benzocyclohepten-5-ols 4a-c and 5a-c and cis- and trans-4-amino 2,3,4,5-tetrahydro-1-benzoxepin-5-ols 4d-f and 5d-f were carried out. Chemo- and stereoselective synthesis of 5a-f was achieved by reduction of corresponding alpha-amino ketones 3a-f with LiAl(t-BuO)3H. cis-4-Amino-2,3,4,5-tetrahydro-1 benzoxepin-5-ol 4d and trans-4-amino-2,3,4,5-tetrahydro-1-benzoxepin-5-ol 5d exhibited marked anorexigenic activity in mice at a dose of LD50 800 and 500 mg/kg and ED50 75 and 55 mg/kg, respectively, while the analog cis-2,3-dihydroxy 6-amino-6,7,8,9-tetrahydro-5H-benzocyclohepten-5-ol 8 showed typical alpha sympathomimetic activity. PMID- 15125950 TI - 3,4-Dihydronaphthalen-1(2H)-ones: novel ligands for the benzodiazepine site of alpha5-containing GABAA receptors. AB - A series of substituted 3,4-dihydronaphthalen-1(2H)-ones with high binding affinity for the benzodiazepine site of GABAA receptors containing the alpha5 subunit has been identified. These compounds have consistently higher binding affinity for the GABAA alpha5 receptor subtype over the other benzodiazepine sensitive GABAA receptor subtypes (alpha1, alpha2 and alpha3). Compounds with a range of efficacies for the benzodiazepine site of alpha5-containing GABAA receptors were identified, including the alpha5 inverse agonist 3,3-dimethyl-8 methylthio-5-(pyridin-2-yl)-3,4-dihydronaphthalen-1(2H)-one 22 and the alpha5 agonist 8-ethylthio-3-methyl-5-(1-oxidopyridin-2-yl)-3,4-dihydronaphthalen-1(2H) one 19. PMID- 15125951 TI - Antimelanomal activity of the copper(II) complexes of 1-substituted 5-amino imidazole ligands against B16F10 mouse melanoma cells. AB - The copper complexes of 5-amino-imidazole ligands were prepared and characterized by various spectroscopic techniques. The ligand geometry around the copper(II) centre is square pyramidal based on N2O2 donor atoms and a coordinated water molecule at the apex. Single crystal X-ray structures were determined for both ligands. Ligands and copper complexes exhibited dose-dependent antiproliferative effects on the growth of B16F10 melanoma cells line but lower IC50 values were observed for the copper complexes. PMID- 15125952 TI - Synthesis of jaspaquinol and effect on viability of normal and malignant bladder epithelial cell lines. AB - The synthesis of jaspaquinol 1, a monocyclic diterpene-benzenoid, is reported. Two synthetic routes to this natural product have been developed. The first, utilises a difunctional terpene derivative containing different leaving groups, facilitating the selective introduction of the cyclohexenyl and benzenoid fragments. The alternative route employs a regiospecific Stille cross-coupling reaction to introduce the cyclohexenyl fragment, which occurs without allylic transposition. Preliminary data shows the cell viability of 1 against normal and malignant human bladder epithelial cell lines. PMID- 15125953 TI - Synthesis and biological activity of novel platinum(II) complexes of glutamate tethered to hydrophilic hematoporphyrin derivatives. AB - A new series of hematoporphyrin-platinum(II) conjugates was prepared by platination of the glutamate ligand tethered to hydrophilic hematoporphyrin derivatives, in which different numbers of ethylene oxide unit were introduced to modulate the hydrophobic/hydrophilic balance of the conjugates. The antitumor activity of the hematoporphyrin-platinum(II) conjugates was assayed in vitro and in vivo against the leukemia L1210 cell line. Among the complexes, compound 11 exhibited not only higher in vivo activity (T/C% = 192) than cisplatin (T/C% = 184) and carboplatin (T/C% = 168), but also elevated tumor-localizing effect (tumor/muscle ratio > 3). PMID- 15125954 TI - Structure-activity relationships of trans-cinnamic acid derivatives on alpha glucosidase inhibition. AB - trans-Cinnamic acid and its derivatives were investigated for the alpha glucosidase inhibitory activity. 4-Methoxy-trans-cinnamic acid and 4-methoxy trans-cinnamic acid ethyl ester showed the highest potent inhibitory activity among those of trans-cinnamic acid derivatives. The presence of substituents at 4 position in trans-cinnamic acid altered the alpha-glucosidase inhibitory activity. Increasing of bulkiness and the chain length of 4-alkoxy substituents as well as the increasing of the electron withdrawing group have been shown to decrease the inhibitory activity. 4-Methoxy-trans-cinnamic acid was a noncompetitive inhibitor for alpha-glucosidase, whereas, 4-methoxy-trans-cinnamic acid ethyl ester was a competitive inhibitor. These results indicated that trans cinnamic acid derivatives could be classified as a new group of alpha-glucosidase inhibitors. PMID- 15125955 TI - Reverse hydroxamate-based selective TACE inhibitors. AB - Reverse hydroxamate-based selective TACE inhibitors are described. They have potent TACE inhibitory activities and excellent selectivities against MMP-1, 2, 3, 8, 9, 13, 14, and 17. One representative compound, 18 has demonstrated an excellent oral inhibitory activity of the lipopolysaccharide (LPS)-stimulated TNF alpha production in rats. PMID- 15125956 TI - Synthesis and evaluation of novel 1,4-naphthoquinone derivatives as antiviral, antifungal and anticancer agents. AB - The synthesis and evaluation of some 2-substituted-1,4-naphthoquinones 2, S-(1,4 naphthoquinon-2-yl)-mercaptoalkanoic acid amides 4, related benzoquinone and naphthoquinone derivatives 6-9 and 2,3-disubstituted 1,4-naphthoquinones 10-11 were carried out. The antifungal, antibacterial, antiviral and anticancer activities were determined by using the standard assay. The results show that compounds 2b and 10a showed in vitro antiviral activity against Influenza-A Virus and Herpes Simplex Virus and possess pronounced antifungal profile whereas 4a showed anticancer activities against Lymphoid Leukaemia P 388. PMID- 15125957 TI - N-Aryl-gamma-lactams as integrin alphavbeta3 antagonists. AB - Novel alphavbeta3 antagonists based on the N-aryl-gamma-lactam scaffold were prepared. SAR studies led to the identification of potent antagonists for alphavbeta3 receptor with excellent selectivity against the structurally related alpha(IIb)beta3 receptor. Additional interactions of N-aryl-gamma-lactam derivatives with alphavbeta3 were found when compared to c(-RGDf[NMe]V-) peptide antagonist. The effects of the conformation and configuration of the gamma-lactam core on the binding were also assessed. PMID- 15125958 TI - Synthesis and biological activity of some structural modifications of pancratistatin. AB - Structurally modified derivatives of 7-deoxypancratistatin have been synthesized and evaluated in cancer cell line inhibition studies. PMID- 15125959 TI - Synthesis of 3a,4-dihydro-3H-[1]benzopyrano[4,3-c]isoxazoles, displaying combined 5-HT uptake inhibiting and alpha2-adrenoceptor antagonistic activities. Part 2: Further exploration on the cinnamyl moiety. AB - In our previous paper we have described the synthesis of a series of 3 piperazinylmethyl-3a,4-dihydro-3H-[1]benzopyrano[4,3-c]isoxazoles, as novel dual 5-HT reuptake inhibitors and alpha2-adrenoceptor antagonists. That investigation led to the identification of the cinnamyl fragment as the most suitable moiety for combined activity. This paper outlines a further optimisation programme, focused on the exploration of the aromatic ring present on the cinnamyl moiety of compounds 1, 2 and 3. PMID- 15125960 TI - New highly selective inhibitors of class II fructose-1,6-bisphosphate aldolases. AB - Phosphoglycolo amidoxime and phosphoglycolo hydrazide, two new derivatives of phosphoglycolic acid, were synthesised and successfully tested as selective competitive inhibitors of class II FBP-aldolases. PMID- 15125961 TI - Synthesis and DNA binding properties of dioxime-peptide nucleic acids. AB - Peptide nucleic acids (PNAs) C- or N-modified with dioxime ligands were prepared by solid-phase synthesis using iron(II)-clathrochelates as protected dioxime building blocks. These PNA bind complementary DNA sequence specifically, though with much reduced affinity in comparison with nonmodified PNA. The dioxime-PNA conjugates bind Cu2+ and Ni2+ at microM concentration. PMID- 15125962 TI - Plasmepsin II inhibition and antiplasmodial activity of Primaquine-Statine 'double-drugs'. AB - Statine-based inhibitors of Plasmepsin II (PLMII) coupled with Primaquine have been designed using the 'double-drug' approach. The IC50 values for PLMII inhibition ranged from 0.59 to 400 nM and the best IC50 value for inhibition of Plasmodium falciparum growth in vitro was 0.4 microM, which represent a remarkable improvement compared to other statine-based PLMII inhibitors. PMID- 15125963 TI - Orally active factor Xa inhibitors: 4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine derivatives. AB - In our investigation of factor Xa inhibitors, a series of 1-(6-chloronaphthalen-2 yl)sulfonyl-4-(4,5,6,7-tetrahydrothiazolo[5,4-c]pyridine-2-carbonyl)piperazines 3a-i were synthesized. In vitro inhibitory activities of the compounds against factor Xa and coagulation are summarized. Among the compounds, 3c and 3d, possessing a carbamoyl or N-methylcarbamoyl moiety, showed potent inhibitory activities when administered orally to rats. PMID- 15125964 TI - The discovery of N-(1,3-thiazol-2-yl)pyridin-2-amines as potent inhibitors of KDR kinase. AB - An azo-dye lead was modified to a novel N-(1,3-thiazol-2-yl)pyridin-2-amine series of KDR kinase inhibitors through the use of rapid analog libraries. This new class has been found to be potent, selective, and of low molecular weight. Molecular modeling has postulated an interesting conformational preference and binding mode for these compounds in the active site of the enzyme. PMID- 15125965 TI - Synergetic inhibition of genistein and D-glucose on alpha-glucosidase. AB - Synergetic inhibitory effect of genistein (I2) and D-glucose (I1) on alpha glucosidase has been studied with kinetics method. It was concluded that the inhibitory effect was much greater when I2 and I1 were both added to the reactant solution simultaneously than that they were added, respectively, which suggesting the inhibitors bind to the different sites of alpha-glucosidase at the same time, and demonstrating synergetic inhibition. PMID- 15125966 TI - Design, synthesis, and characterization of an ATP-peptide conjugate inhibitor of protein kinase A. AB - An ATP-peptide conjugate was synthesized as a bisubstrate analogue inhibitor of the serine/threonine kinase protein kinase A. The compound was found to be a linear, competitive inhibitor with respect to ATP substrate, exhibiting a Ki of 3.8 microM. The compound was noncompetitive with respect to peptide substrate. The inhibitor was shown to be selective for protein kinase A versus the closely related protein kinase C as well as tyrosine kinase Csk. This analysis provides new evidence for the dissociative transition state of protein serine/threonine kinases and illustrates a simple method to convert a low affinity peptide substrate to a selective and moderately potent inhibitor for these enzymes. PMID- 15125967 TI - Identification of purine inhibitors of phosphodiesterase 7 (PDE7). AB - A series of purine based inhibitors of PDE7 has been derived from screening lead 1a. The synthesis, structure-activity relationships (SAR), and selectivity against several other PDE family members are described. PMID- 15125968 TI - Synthesis and DNA damaging ability of enediyne model compounds possessing photo triggering devices. AB - Enediyne model compounds possessing photo-triggering devices were developed. These enediynes afforded biradicals by UV irradiation and showed DNA cleaving activity. The DNA damage was confirmed to be mainly caused by the biradical, not singlet oxygen. PMID- 15125969 TI - Design, synthesis, and structure-activity relationship of new isobenzofuranone ligands of protein kinase C. AB - Protein kinase C (PKC) is a family of enzymes, which play important roles in intracellular signal transduction. We have designed novel PKC ligands having an isobenzofuranone template, based on the proposed interaction of DAG (1,2-diacyl sn-glycerol) with the PKCdelta C1B ligand-binding domain. Several isobenzofuranone derivatives were synthesized and their PKCalpha binding activities were evaluated. The pivaloyl derivative 1f was found to be a strong PKCalpha ligand, and the structure-activity relationship is well explained by our proposed binding model. PMID- 15125970 TI - Evaluation of series of isobenzofuranone dimers as PKCalpha ligands: implication for the distance between the two ligand binding sites. AB - Protein kinase C (PKC) is a family of enzymes, which play important roles in intracellular signal transduction. To examine the distance between the two ligand binding sites (C1A and C1B) of PKC, we designed and synthesized two series of isobenzofuranone dimers. Peak binding activities were observed for the C3-acyl chain dimers having a C10-C12 linker and for the C7 dimers having a C14-C16. PMID- 15125971 TI - Synthesis and biological activity of N-aryl-2-aminothiazoles: potent pan inhibitors of cyclin-dependent kinases. AB - N-Aryl aminothiazoles 6-9 were prepared from 2-bromothiazole 5 and found to be CDK inhibitors. In cells they act as potent cytotoxic agents. Selectivity for CDK1, CDK2, and CDK4 was dependent of the nature of the N-aryl group and distinct from the CDK2 selective N-acyl analogues. The N-2-pyridyl analogues 7 and 19 showed pan CDK inhibitory activity. Elaborated analogues 19 and 23 exhibited anticancer activity in mice against P388 murine leukemia. The solid-state structure of 7 bound to CDK2 shows a similar binding mode to the N-acyl analogues. PMID- 15125972 TI - Antitumor agents. Part 230: C4'-esters of GL-331 as cytotoxic agents and DNA topoisomerase II inhibitors. AB - Three C4'-acyl derivatives (5-7) of GL-331 (4) were synthesized and evaluated for cytotoxic and DNA topoisomerase II (topo II) inhibitory activity. All three compounds were cytotoxic against KB and KB-7d cells. Compounds 5 and 7, but not 6, were potent inhibitors of the DNA topoisomerase II in vitro and this relative activity ranking was maintained for cytotoxicity, in vitro cell growth inhibition, and ability to induce cellular double-strand DNA breaks. These results provided new information on the structure-activity relationships of the C4' molecular area of 4-analogues. PMID- 15125973 TI - Synthesis of dammarane-type triterpenoids with anti-inflammatory activity in vivo. AB - The 17-alpha-substituted triterpene 1 [(17alpha)-23-(E)-dammara-20,23-diene 3beta,25-diol] showed promising activity in animal models of immunosuppression and inflammation. Using a mouse model for inflammatory skin diseases (oxazolone induced allergic contact dermatitis, ACD) as the directing in vivo test system, Structure-activity-relationship studies with the aim to understand the necessary structural requirements for the biological activity of 1 were conducted. Furthermore, we anticipated to identify biologically active compounds with the 17beta configuration, which are thermodynamically more stable and much easier to synthesize. This was achieved by identifying the 17-beta substituted dammarane 5B and its analogues. PMID- 15125974 TI - Synthesis and evaluation of spirobenzazepines as potent vasopressin receptor antagonists. AB - A novel series of spirobenzazepines was synthesized and evaluated for V1a and V2 receptor antagonist activity. Compounds 8b, 8i, and 8k have shown selective V1a receptor antagonist activity. Compounds 8p and 8q were shown to be dual V1a/V2 receptor antagonists. PMID- 15125975 TI - Normative development of fears in Greece: self-reports on the Hellenic Fear Survey Schedule for Children. AB - This study assessed the psychometric properties of self-reports on the Hellenic Fear Survey Schedule for Children (FSSC-GR), a Greek-language version of Ollendick's FSSC-R [Behav. Res. Ther. 21 (1983) 685-692], and explored the factor structure, intensity, prevalence, and content of fears of Hellenic children aged 7-12 years. Internal consistency and temporal stability assessments of responding on the FSSC-GR were comparable to those obtained on the FSSC-R. A seven-factor solution provided the best conceptual fit for the structure of children's fears in Greece, including five components similar to factors previously obtained for the FSSC-R, plus two unique components, "Travel and Agoraphobic Fears" and "School Performance Fears." Fear intensity and prevalence scores of Hellenic children were higher than scores observed in most countries. Self-reported fear scores were higher for Hellenic girls than for boys. The relation between age and self-reported fears in Greece was complex and interpreted in relation to age related changes in environmental demands. PMID- 15125976 TI - Eating disorder symptoms among female anxiety disorder patients in clinical practice: the importance of anxiety comorbidity assessment. AB - This exploratory study investigated the relationship between anxiety disorders, anxiety comorbidity, and eating disorder (ED) symptoms in clinical practice, and examined the naturalistic detection of ED when diagnoses were based on the Anxiety Disorders Interview Schedule (ADIS). Two hundred and fifty-seven female patients completed an ED questionnaire and were assessed with the ADIS. Although ED frequency did not differ among anxiety disorder diagnoses, regression analyses revealed that social phobia (SP) and posttraumatic stress disorder (PTSD) accounted for unique variance in eating pathology. Questionnaire results indicated that almost 12% of patients met criteria for a possible ED. Clinicians using the ADIS evidenced good specificity but were not sensitive to detecting ED, missing 80% of possible cases. Results support possible links between ED, social phobia and PTSD and highlight the importance of assessing anxiety comorbidity when examining the relationship between ED and anxiety disorders. Results also suggest that formal screening for ED among female anxiety patients may be warranted. PMID- 15125977 TI - Distinguishing generalized anxiety disorder, panic disorder, and mixed anxiety states in older treatment-seeking adults. AB - Eighty treatment-seeking adults age 60 or over with panic disorder, generalized anxiety disorder, and mixed anxiety states (generalized anxiety with panic attacks, panic disorder with secondary generalized anxiety) completed a clinical assessment and battery of self report measures. Several hypotheses were tested from the domains of distinguishing symptoms, associated features, and rates of comorbidity with other disorders. Greater between- than within-group variance was found on a subset of measures suggesting that the distinction between GAD and PD is generally valid in the older adult population. Higher scores on measures of sympathetic arousal, agoraphobic avoidance, and rates of comorbid somatization disorder and alcohol dependence distinguished those with PD from those with GAD. Higher scores on measures of depression and hostility, but not trait anxiety or worry, distinguished the GAD group. Results indicate that distinguishing features of GAD and PD in older treatment-seeking adults may be fewer and slightly different from those of younger adults. PMID- 15125978 TI - A study of the psychometric properties of the Obsessive Beliefs Inventory and Interpretations of Intrusions Inventory on clinical Italian individuals. AB - Forty-three individuals with obsessive-compulsive disorder (OCD), 17 with generalized anxiety disorder (GAD) and 50 non-clinical controls (SC), completed the Italian versions of the Obsessive Beliefs Questionnaire (OBQ) and the Interpretations of Intrusions Inventory (III), along with measures of obsessive compulsive (OC) symptoms, depression, anxiety and worry. OBQ and III showed an excellent reliability and temporal stability. The six OBQ subscales were reasonably distinct from each other, whereas the three subscales of the III were highly interrelated. The OBQ, and in less measure the III, discriminated between OCD, GAD, and normal controls. In the OCD cohort, the two instruments correlated moderately with a measure of obsessive-compulsive symptoms but also with a measure of worry. However, a linear regression analysis evidenced a specific relationship between the OBQ and obsessive-compulsive symptoms over and above worry. Overall, at least three cognitive domains (intolerance of uncertainty, excessive concern about the importance of controlling one's thoughts and perfectionism) seemed specific to OCD, whereas overimportance of thoughts and inflated responsibility barely discriminate clinically anxious individuals from non-clinical ones. It is concluded that OBQ and III are useful measures in Italian individuals and that more research is warranted to possibly refine these two instruments. PMID- 15125979 TI - Avoidance-oriented coping as a predictor of panic-related distress: a test using biological challenge. AB - The present experimental psychopathology study sought to investigate the extent to which pre-experimental levels of avoidance-oriented coping predict anxious and fearful responding during acute physical stress, relative to other theoretically relevant variables. Participants included 80 individuals with no known history of psychological or physical health problems. Dependent measures include self reported anxiety, DSM-IV panic symptoms, and physiological indices of heart rate and skin conductance. Consistent with our hypotheses, the tendency to engage in avoidance-oriented coping predicted increased physical panic symptoms and self reported anxiety elicited by biological challenge, relative to specific anxiety sensitivity (AS) dimensions. These findings are discussed in terms of how specific types of coping are associated with prototypical indices of panic distress, with implications for forwarding future work on emotional regulation in panic disorder. PMID- 15125980 TI - The role of catastrophic misinterpretation of bodily sensations and panic self efficacy in predicting panic severity. AB - This study investigated the role of both negative and positive cognitions in predicting panic severity in an international sample of patients diagnosed with panic disorder (with and without agoraphobia). One hundred and fifty-nine patients were administered the Brief Bodily Sensations Interpretation Questionnaire (BBSIQ), the Self-efficacy to Control Panic Attacks Questionnaire, and the Panic and Agoraphobia Scale (PAS) prior to receiving treatment. Regression analyses indicated that both catastrophic misinterpretation of bodily sensations and panic self-efficacy independently predicted panic severity. The influence of panic self-efficacy upon panic severity remained significant even after controlling for the presence or absence of agoraphobia. There was no evidence to suggest a moderating relationship between the two cognitive factors. Results are discussed in terms of the need to consider both negative and positive cognitions in cognitive accounts of panic disorder. PMID- 15125981 TI - Balance dysfunction in childhood anxiety: findings and theoretical approach. AB - A recent special issue of the Journal of Anxiety Disorders, reviewed the experimental and clinical findings related to comorbidity of balance disorders and anxiety [J. Anxiety Disord. 15 (2001) 1.]. The studies mentioned in that issue were based mostly on adult subjects but prevalence of balance disorders in childhood anxiety is yet to be established. We have tested a small sample of children diagnosed for general or separation anxiety disorder and a control group of normal children. Extensive neurological examination revealed no clinically relevant vestibular impairment. Nevertheless, detailed questionnaires and balance tests confirmed an excessive sensitivity of anxiety disordered children to balance-challenging situations. Moreover, balance-challenging tasks triggered more balance mistakes and slower performance in anxiety versus control children. These findings support the notion of subclinical balance disorder in childhood anxiety. Results are discussed in terms of the two-stage theory of learning, which predicts that anxiety disorder may be an offshoot of lasting balance dysfunction. PMID- 15125982 TI - Cognitive-behavioral treatment of tortured asylum seekers: a case study. AB - The present study examined results of cognitive-behavioral treatment (CBT) in a 22-year-old, male, tortured asylum-seeker living in Sweden. The patient received 16 sessions of CBT involving mainly self-exposure to trauma-related cues. Clinical measures (assessor- and self-rated) were completed at pre-treatment, weeks 6, 8, 12, and 16, post-treatment and at follow-up (1-, 3-, and 6-month). Treatment led to significant improvement across all measures of post-traumatic stress disorder, anxiety, and depression. The improvement was maintained at 6 month follow-up. The results suggest that CBT could be useful in treating tortured asylum-seekers and refugees despite the additional stressors experienced by asylum-seekers and refugees. PMID- 15125983 TI - The path from initial inquiry to initiation of treatment for social anxiety disorder in an anxiety disorders specialty clinic. AB - Efficacious treatments for social anxiety disorder have become increasingly available, with approximately three-quarters of treatment completers showing significant improvement [Arch. Gen. Psychiatry 55 (1998) 1133.]. However, very few individuals with social anxiety disorder access these services. The present study reports on the path to initiation of treatment for social anxiety disorder among individuals contacting an anxiety disorder specialty clinic. Of 395 initial telephone inquiries, only 60 individuals (15%) started treatment. Three "critical points" associated with high pretreatment attrition were identified: scheduling an initial interview, attending a scheduled initial interview, and initiating a treatment program after receiving a principal diagnosis of social anxiety disorder. Several demographic variables (e.g., level of education, race) were related to attendance at the initial interview. However, no differences were found between those who did and did not initiate treatment on demographic variables, symptom severity, or quality of life. Given that most individuals who complete treatment for social phobia experience significant benefit, results of the current study suggest that future efforts should be devoted to increasing number of patients who access these services. PMID- 15125984 TI - Predictors of fear of crime in older adults. AB - Very little is known about factors that predict fear of crime in older adults. Indeed, the topic itself remains a source of controversy, with early studies indicating higher levels of crime fear with age, and new, more methodologically rigorous studies demonstrating the opposite trend. The present exploratory investigation included 106 older adults and assessed the relationship between demographic variables, interpersonal violence, psychopathology, and fear of crime. In addition, this study built on previous research in that specific feared outcomes (e.g., hospitalization) were also considered. Initial findings indicate that being female, non-Caucasian, having depressive symptoms, and reporting social isolation are predictive of general fear of crime ratings. Different predictor sets were noted for fear of crime against person and fear of crime against property. Reported perceptions of negative crime outcomes were associated with being female, non-Caucasian, and having low income. PMID- 15125985 TI - Modification and psychometric examination of a self-report measure of fear in older adults. AB - The primary purposes of the present study were to: (a) develop a measure of self reported fear that includes fears that are content valid for older adults, (b) develop an instrument for assessing daily interference associated with fears, and (c) examine the psychometric properties of this new instrument. In the first of two studies, 109 participants, ages 60 through 91, completed a demographic questionnaire and answered an open-ended question about the types of fears they experience. These fears were then included on a modified version of the Fear Survey Schedule-II (Fear Survey Schedule-II for Older Adults (FSS-II-OA)) and administered in a second study. In Study 2, 114 participants (ages 60 through 88) completed a demographic questionnaire, FSS-II-OA, Beck Anxiety Inventory (BAI), Beck Depression Inventory-II (BDI-II), and Penn State Worry Questionnaire (PSWQ), and answered questions regarding fear avoidance. Study 2 revealed high levels of internal consistency and test-retest reliability for total scores on the FSS-II OA. Test-retest reliability estimates were mixed for individual fear intensity and daily interference items on the FSS-II-OA. Low-moderate to moderate correlations were found between the FSS-II-OA and BAI, BDI-II, and PSWQ. PMID- 15125986 TI - Development and preliminary validation of the Meta-cognitions Questionnaire Adolescent Version. AB - This study describes the development of the Meta-cognitions Questionnaire for Adolescents (MCQ-A). The adult version of this questionnaire has been widely used, and the meta-cognitive theory of emotional disorders has become influential in the adult literature. This study sought to examine the prevalence and emotional correlates of meta-cognition in adolescents. The MCQ-A was administered to 177 schoolchildren aged 13 through 17 years, together with measures of emotional well-being. Factor analysis suggested a five-factor solution similar to that reported for the adult version. Internal consistency of the scale was high. Scores ranged from low to high across the age range, suggesting that meta cognitive beliefs are well-established by adolescence. Scores on the MCQ-A were highly correlated with a number of measures of emotional symptoms. The MCQ-A is a reliable and valid instrument for measuring meta-cognitive beliefs in adolescents. Young people aged between 13 and 17 years report a range of types and levels of meta-cognitive beliefs and these are positively associated with emotional symptoms. PMID- 15125987 TI - Anxiety sensitivity and modulation of the serotonergic system in patients with PD. AB - Anxiety sensitivity, i.e., the fear of anxiety-related bodily sensations, is one of the most studied cognitive variables in panic disorder (PD). However, the effects of selective serotonergic antipanic agents on this variable have not yet been investigated. The present study examines the effects of 6 weeks of treatment with citalopram on anxiety sensitivity in patients with PD. Twenty patients entered the study. On day 0, before starting drug treatment, after 1 week and after 6 weeks of treatment, each patient was evaluated with the Anxiety Sensitivity Index (ASI); the severity of clinical symptomatology was assessed with standardized psychometric scales. Results showed a significant reduction of anxiety sensitivity after 6 weeks of treatment. There was a significant correlation between decrease of anxiety sensitivity and anticipatory anxiety, while no correlations were found between panic attacks and agoraphobic avoidance. These results suggest that antipanic drug treatment decreases anxiety sensitivity. PMID- 15125988 TI - Multiset splicing systems. AB - We consider splicing systems reflecting two important aspects of the behaviour of DNA molecules in nature or in laboratory experiments which so far have not been studied in the literature. We examine the effect of splicing rules applied to finite multisets of words using sequential and different types of parallel derivation strategies and compare the sets of words or sets of multisets which can be obtained. PMID- 15125989 TI - DNA computation model to solve 0-1 programming problem. AB - 0-1 programming problem is an important problem in opsearch with very widespread applications. In this paper, a new DNA computation model utilizing solution-based and surface-based methods is presented to solve the 0-1 programming problem. This model contains the major benefits of both solution-based and surface-based methods; including vast parallelism, extraordinary information density and ease of operation. The result, verified by biological experimentation, revealed the potential of DNA computation in solving complex programming problem. PMID- 15125990 TI - Mutation and selection in a large population. AB - In this paper we study a large, but finite population, in which mutation and selection occur at a single genetic locus in a diploid organism. We provide theoretical results for the equilibrium allele frequencies, their variances and covariances and their equilibrium distribution, when the population size is larger than the reciprocal of the mean allelic mutation rate. We are also able to infer that the equilibrium distribution of allele frequencies takes the form of a constrained multivariate Gaussian distribution. Our results provide a rapid way of obtaining useful information in the case of complex mutation and selection schemes when the population size is large. We present numerical simulations to test the applicability of our theoretical formulations. The results of these simulations are in very reasonable agreement with the theoretical predictions. PMID- 15125991 TI - Learning and evolution in bacterial taxis: an operational amplifier circuit modeling the computational dynamics of the prokaryotic 'two component system' protein network. AB - Adaptive behavior in unicellular organisms (i.e., bacteria) depends on highly organized networks of proteins governing purposefully the myriad of molecular processes occurring within the cellular system. For instance, bacteria are able to explore the environment within which they develop by utilizing the motility of their flagellar system as well as a sophisticated biochemical navigation system that samples the environmental conditions surrounding the cell, searching for nutrients or moving away from toxic substances or dangerous physical conditions. In this paper we discuss how proteins of the intervening signal transduction network could be modeled as artificial neurons, simulating the dynamical aspects of the bacterial taxis. The model is based on the assumption that, in some important aspects, proteins can be considered as processing elements or McCulloch Pitts artificial neurons that transfer and process information from the bacterium's membrane surface to the flagellar motor. This simulation of bacterial taxis has been carried out on a hardware realization of a McCulloch-Pitts artificial neuron using an operational amplifier. Based on the behavior of the operational amplifier we produce a model of the interaction between CheY and FliM, elements of the prokaryotic two component system controlling chemotaxis, as well as a simulation of learning and evolution processes in bacterial taxis. On the one side, our simulation results indicate that, computationally, these protein 'switches' are similar to McCulloch-Pitts artificial neurons, suggesting a bridge between evolution and learning in dynamical systems at cellular and molecular levels and the evolutive hardware approach. On the other side, important protein 'tactilizing' properties are not tapped by the model, and this suggests further complexity steps to explore in the approach to biological molecular computing. PMID- 15125992 TI - Gene duplication and hierarchical modularity in intracellular interaction networks. AB - Networks of interactions evolve in many different domains. They tend to have topological characteristics in common, possibly due to common factors in the way the networks grow and develop. It has been recently suggested that one such common characteristic is the presence of a hierarchically modular organization. In this paper, we describe a new algorithm for the detection and quantification of hierarchical modularity, and demonstrate that the yeast protein-protein interaction network does have a hierarchically modular organization. We further show that such organization is evident in artificial networks produced by computational evolution using a gene duplication operator, but not in those developing via preferential attachment of new nodes to highly connected existing nodes. PMID- 15125993 TI - Emergence of power laws from partitional dynamics. AB - This work explores the possibility of producing some specific power laws (with exponential cut-off) out from a partitional dynamics. The data obtained from the partitions of integers in the interval 1-60 have been used to find the coefficients of a power law and its exponential cut-off, and also of a single hyperbolic form after renormalization. There is also the speculation that this type of power law, so easily derived from arithmetic minimalist operations, may underlie the communication exchanges of living cells and the structural games of self-constructing agents endowed with relative structural freedom. PMID- 15125994 TI - An approach to measuring change in muscular tissue contraction. AB - In this paper the authors consider a histological muscular component from the point of view of systems theory. Of the smooth and striated types of muscular fibres, the latter have a structural component that permits them to contract. Unlike the visceral striated and cardiac fibres, movements in the skeletal striated fibres are predominantly voluntary. An overlapping of certain fibre components--specifically a sliding movement of the filaments of which the fibre is composed--causes a variation in the overall size of the fibre and therefore of the muscle. The distances covered, whether in contraction or lengthening, vary according to the type of fibre from 1.8 to 3.8 microm, but depend only on the capacity of some filaments to slide over other filaments, with filament size itself remaining unchanged. Our objective is to obtain a metric of this system that will permit us to obtain some idea of the degree of variation in these distances. In future projects it will be possible to extend this theory to other involuntary muscular tissue movements (myocardial tissue, e.g.), in an endeavour to develop a mathematical approach to the study of myocardial processes damaged by electrical conduction problems. PMID- 15125995 TI - Inhalation toxicology and carcinogenesis studies of propylene glycol mono-t-butyl ether in rats and mice. AB - Propylene glycol mono-t-butyl ether (PGMBE) is used as a solvent in a variety of commercial applications. Male and female F344/N rats and B6C3F(1) mice were exposed to PGMBE by whole-body inhalation for 2 or 14 weeks (0, 75, 150, 300, 600, or 1200 ppm) or 2 years (0, 75, 300, or 1200 ppm); male NBR rats were exposed for 2 weeks. The kidney and the liver were targets of PGMBE toxicity in rats. Renal lesions suggestive of alpha(2u)-globulin nephropathy were observed in male F344/N, in the 2 and 14-week studies, no kidney lesions were seen in NBR rats. In the 2-year study, male rats displayed exposure-related nonneoplastic lesions in the kidney, and may have shown marginal increases in tubular neoplasms. In the liver, the incidences of hepatocellular adenomas occurred with a positive trend in male rats, and may have been related to PGMBE exposure. In mice of both sexes, the major target of PGMBE toxicity was the liver. In the 2 week study, liver weights and in the 14-week study, liver weights and the incidences of centrilobular hypertrophy were increased. In the 2-year study, the incidences of exposure-related hepatocellular adenoma, adenoma or carcinoma combined, and hepatoblastoma occurred with a positive trend, and were significantly increased in 1200 ppm groups. In summary, exposure to PGMBE resulted in nonneoplastic lesions of the kidney characteristic of alpha(2u) globulin nephropathy, and may have increased renal tubular neoplasms in male rats. Exposure to PGMBE also produced increases in hepatic tumors in male and female mice. PMID- 15125996 TI - A human and mouse pregnane X receptor reporter gene assay in combination with cytotoxicity measurements as a tool to evaluate species-specific CYP3A induction. AB - Recent studies have demonstrated that a member of the nuclear receptor family, pregnane X receptor (PXR) is a key regulator of the expression of cytochrome P450 3A (CYP3A) in humans and rodents. It is also known that species specificity in the induction of CYP3A by xenobiotics is likely a consequence of differences at the level of PXR activation. Because of the importance of CYP3A4 in drug metabolism, the development of rapid and accurate in vitro assays for predicting the effects of compounds on CYP3A4 expression or activity in humans has been a long-standing goal within pharmaceutical industries. PXR activation measurements using an in vitro reporter gene approach appears to provide a rapid and relatively inexpensive means for predicting whether compounds will induce CYP3A levels in vivo. In this study, using an HepG2 cell based human and mouse PXR reporter gene assay, 23 compounds were tested for their potential to activate hPXR or mPXR. Data demonstrated that potent activators of hPXR had virtually no activity on mPXR and efficient activators of mPXR had weak activity on hPXR. In addition, a third category of moderate/weak activators of both hPXR and mPXR was identified. Exemestane was a strong activator of mPXR ( approximately 22-fold activation) with only minor effect on hPXR ( approximately 5-fold activation). The importance of cell viability measurements as part of the PXR reporter gene assay was demonstrated as significant cytotoxicity or inhibition of cell proliferation might underestimate the potential for PXR activation. PMID- 15125997 TI - Benzo(a)pyrene-induced apoptotic death of mouse hepatoma Hepa1c1c7 cells via activation of intrinsic caspase cascade and mitochondrial dysfunction. AB - Benzo(a)pyrene (BaP), a potent carcinogen, has been shown to induce apoptosis via activation of caspase-3. However, the upstream of caspase-3 and other apoptosis signaling remain to be elusive. Herein, we demonstrated that treatment of Hepa1c1c7 cells with BaP increased the transcriptional expression of aryl hydrocarbon nuclear transporter and cytochrome p450 1A1 in a time and dose dependent manner but did not aromatic hydrocarbon receptor. Also, the catalytic activation of caspase-3 and caspase-9 was induced whereas that of caspase-3 and caspase-9 was not by the addition of BaP. BaP also induced the mitochondrial dysfunction, including transition of mitochondria membrane potential and cytosolic release of cytochrome c. Furthermore, a decrease in the expression of Bcl-2 to Bax ratio and phosphorylation of p53(Ser 15) were observed in BaP treated cells. Taken together, these results demonstrated that BaP-induced apoptosis of Hepa1c1c7 cells via activation of intrinsic caspase pathway including caspase-3, caspase-9, with mitochondrial dysfunction and p53 activation. PMID- 15125998 TI - Aluminium-induced changes in hemato-biochemical parameters, lipid peroxidation and enzyme activities of male rabbits: protective role of ascorbic acid. AB - For a long time, aluminium (Al) has been considered an indifferent element from a toxicological point of view. In recent years, however, Al has been implicated in the pathogenesis of several clinical disorders, such as dialysis dementia, the fulminant neurological disorder that can develop in patients on renal dialysis. Therefore, the present experiment was carried out to determine the effectiveness of l-ascorbic acid (AA) in alleviating the toxicity of aluminium chloride (AlCl3) on certain hemato-biochemical parameters, lipid peroxidation and enzyme activities of male New Zealand white rabbits. Six rabbits per group were assigned to 1 of 4 treatment groups: 0mg AA and 0mg AlCl3/kg body weight (BW) (control); 40 mg AA/kg BW; 34 mg AlCl3/kg BW (1/25 LD50); 34 mg AlCl3 plus 40 mg AA/kg BW. Rabbits were orally administered their respective doses every other day for 16 weeks. Evaluations were made for lipid peroxidation, enzyme activities and hemato biochemical parameters. Results obtained showed that AlCl3 significantly (P<0.05) induced free radicals and decreased the activity of glutathione S-transferase (GST) and the levels of sulfhydryl groups (SH groups) in rabbit plasma, liver, brain, testes and kidney. Aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (AlP), acid phosphatase (AcP), and phosphorylase activities were significantly decreased in liver and testes due to AlCl3 administration. While, plasma, liver, testes and brain lactate dehydrogenase (LDH) activities were significantly increased. Contrariwise, the activity of acetylcholinesterase (AChE) was significantly decreased in brain and plasma. Aluminium treatment caused a significant decrease in plasma total lipids (TL), blood haemoglobin (Hb), total erythrocytic count (TEC) and packed cell volume (PCV), and increased total leukocyte count (TLC) and the concentrations of glucose, urea, creatinine, bilirubin and cholesterol. Ascorbic acid alone significantly decreased the levels of free radicals, TL, cholesterol, glucose and creatinine, and increased the activity of GST, SH groups, Hb, TEC and PCV. While, the rest of the tested parameters were not affected. Also, the present study showed that ascorbic acid can be effective in the protection of aluminium-induced toxicity. PMID- 15125999 TI - Disruption of mice sleep stages induced by low doses of organophosphorus compound soman. AB - We have explored during 7 days, EEG spectral response and sleep pattern of mice after a mild intoxication with soman. Using an automatic staging method, we have quantified the sleep stage of the mice to identify disruptions of the sleep pattern. The 50 microg/kg dose of soman produced several effects during several time windows after intoxication. A first decrease followed by an increase of theta energy, a disturbance of slow wave sleep during 5 days and an increase of the REM sleep during the first and second day after intoxication. During the first 6h, we have observed some effects which were not consistent with a muscarinic activation and might have involved GABA-ergic system. After this early period, the observed effects were in accordance with a muscarinic activation. We observed an increase of energy in the EEG theta band during 3 days after soman injection and an increase of slow wave sleep during the second to the fifth day after soman injection. PMID- 15126000 TI - Nasal allergy-like symptoms aggravated by ozone exposure in a concentration dependent manner in guinea pigs. AB - Our previous study revealed that exposure to 0.4 ppm ozone (O(3)) enhanced nasal allergy-like reactions in guinea pigs. In the present study, we investigated the concentration-dependency of the effects of exposure to O(3) on the aggravation of nasal allergy-like reactions induced by repeated nasal administration of antigen. Guinea pigs were exposed to filtered air or 0.1-0.6 ppm O(3) for 5 weeks. After each weekly administration of ovalbumin (OVA), sneezes and nasal secretions were measured. The number of eosinophils infiltrating the nasal septum and the titers of OVA-specific antibody were measured 24h after the last administration. Ozone increased sneezing and nasal secretion induced by OVA, nasal responsiveness to physical stimuli, and the number of infiltrating eosinophils in a concentration dependent manner. The titer of anti-OVA-IgG was increased in animals exposed to 0.6 ppm O(3). Thus, exposure to O(3) aggravated nasal allergy-like symptoms concentration dependently. The aggravation was caused by induction of nasal hyperresponsiveness, the infiltration of eosinophils, and the increase in the production of anti-OVA-IgG. The estimated maximum likelihood estimation concentrations (MLECs) and bench mark concentrations (BMCs) of O(3) for these indices were in the range of 0.09-0.18 and 0.02-0.06 ppm, respectively. PMID- 15126001 TI - Prostaglandin E1 in lipid microspheres ameliorates diabetic peripheral neuropathy: clinical usefulness of Semmes-Weinstein monofilaments for evaluating diabetic sensory abnormality. AB - We investigated the effect of Prostaglandin E1 in lipid microspheres (Lipo-PGE1) on diabetic peripheral neuropathy from view of symptoms, neurological examinations including sensory threshold evaluated with Semmes-Weinstein monofilaments (SWM). Type 2 diabetic patients with diabetic peripheral neuropathy were participated in this study. The patients were randomly assigned to two groups, 11 Lipo-PGE1-treated patients and 16 control patients. Lipo-PGE1 at a dose of 10mg in 20ml of saline was injected intravenously as a bolus once daily for 2 weeks. Before and, 1, 2 and 4 weeks after the start of treatment with Lipo PGE1, sensory threshold was evaluated with Semmes-Weinstein monofilaments at total 18 touch sites on the feet. Administration of Lipo-PGE1 improved subjective symptoms especially in items of numbness and imperception. Such improvement in subjective symptoms correlated well with the improvement in Semmes-Weinstein monofilaments examination, whereas the improvement was not recognized in motor nerve conduction velocity (MCV), sensory nerve conduction velocity (SCV) and coefficient variation of R-R interval on ECG (CVR-R). The improvement lasted for at least 6 months. This study demonstrated that Lipo-PGE1 has long term amelioration effects on diabetic neuropathy especially in symptoms and sensory threshold, and that Semmes-Weinstein monofilaments examination is a simpler, more valid and quantitative tool for assessing the clinical effect of Lipo-PGE1 on diabetic peripheral neuropathy. PMID- 15126002 TI - Quantitative ultrasound of proximal phalanxes in patients with type 1 diabetes mellitus. AB - The influence of duration of diabetes and metabolic control on phalangeal quantitative ultrasound (QUS) was evaluated in a group of children and adolescents with type 1 diabetes. Eighty-six patients (mean age 11.9 years; mean duration 4.3 years) were studied. Daily calcium intake was assessed by means of a questionnaire. Amplitude-dependent speed of sound (AD-SoS) was measured at the phalanxes of the non-dominant hand and expressed as a z-score. Linear and multivariate correlations with duration of diabetes and, short term and long term metabolic control were sought. AD-SoS z-score was -0.43+/-1.4 (95% CI, -0.73; 0.13). Nine subjects had values below -2S.D. Daily calcium intake was 1042+/-456 mg/day; 47 subjects (54.6%) were below the recommended levels. A negative correlation was found between AD-SoS z-score and duration (r, -0.33, P=0.002) or metabolic control (HbA1c-last year r, -0.32, P=0.002; HbA1c-whole duration, r, 0.40; P=0.003). Negative AD-SoS z-scores depended significantly and directly on duration and quality of metabolic control, even when controlled for calcium intake. In conclusion, the architectural organization of bone was impaired in 10.5% patients. Duration of diabetes and poor metabolic control were the main determinants affecting AD-SoS. QUS may be a useful tool in the screening of bone disturbance in young patients with diabetes. Optimization of metabolic control is required to prevent osteoporosis. PMID- 15126003 TI - The acute effects of in-patient physiotherapy program on functional capacity in type II diabetes mellitus. AB - PURPOSE: This study was planned to evaluate the acute effects of a short-term in patient physiotherapy program on functional capacity in hospitalised type II diabetic patients. METHODS: Forty four hospitalised type II diabetic patients in continuing medical care at Dokuz Eylul University Hospital were included in this study. Twenty three of the 44 patients were randomised to an exercise rehabilitation group and 21 to a control group. Patients in the exercise group were assigned to an average 12.0+/-2.4 days low-intensity exercise rehabilitation program during hospitalisation. The control group did not receive any exercise advice. Before and after the exercise rehabilitation program all patients' functional capacity was evaluated by the 6 min walking test. RESULTS: There were no significant differences in the main measured parameters among groups and in the 6 min walking test at baseline. At the end of the rehabilitation program, the resting heart rate (HR) decreased by 4.1% (P<0.05) in the exercise group by and 2.1% in control group. Resting systolic (SBP) and diastolic blood pressure (DBP) decreased (6.2 and 2.6%) significantly in the exercise group but there were no significant changes in the control group (0.9 and 0.7%). The distance walked in the 6 min test increased significantly more in the exercise group than in the control group ( 18.0% versus 10.2%; P<0.05). Estimated VO2 max also increased more (28.6% versus 3.5%, P<0.05). Moreover, the exercise group described the 6 min walk test as easier than before exercise rehabilitation. The Borg scale did not change in the control group. CONCLUSION: Supervised in-patient exercise rehabilitation is a safe and effective intervention in type II diabetic patients, which reduces physical impairment and improves functional ability. PMID- 15126004 TI - Relationship between visceral fat accumulation and physical fitness in Japanese women. AB - OBJECTIVE: To investigate the link between visceral fat (V) accumulation and physical fitness. DESIGN: Cross-sectional clinical investigation study. SUBJECTS: Twenty-eight overweight Japanese women with abdominal obesity (abdominal obesity) (body mass index, BMI>or=25, visceral fat area>or=100 cm2) aged 38-65 years (BMI 29.5+/-3.3 kg/m2) were enrolled in this study. They were compared with age and sex matched 28 normal weight subjects (BMI<25, visceral fat area<100 cm2) and 28 overweight subjects without abdominal obesity (overweight) (BMI>or=25, visceral fat area<100 cm2). MEASUREMENTS: Fat distribution was evaluated by visceral fat and subcutaneous fat (S) areas measured by computed tomography (CT) scanning at umbilical level. Anthropometric parameters, aerobic exercise level, muscle strength, flexibility and blood examination were also investigated. RESULTS: Anthropometric and body composition parameters were significantly higher in abdominal obesity subjects than those in normal weight subjects. BMI, waist circumference, waist hip ratio and visceral fat (V) area were also significantly higher than those in overweight subjects. Reduced oxygen uptake at ventilatory threshold (VT), lowering weight bearing index (WBI), and elevation of blood enzymes were noted in abdominal obesity subjects. There were significant relationships between oxygen uptake at VT and visceral fat area, and also between oxygen uptake at VT and WBI. CONCLUSION: Reduced aerobic exercise level and WBI are characteristic and visceral fat may play an important role for the etiology in Japanese women with abdominal obesity. PMID- 15126005 TI - Glibenclamide-induced acute haemolytic anaemia revealing a G6PD-deficiency. AB - A 58-year-old woman was admitted at diagnosis of type 2 diabetes without keto acidosis. Blood glucose was normalized initially with insulin. Whilst taking glibenclamide, she developed acute haemolysis. She was homozygous for glucose-6 phosphate dehydrogenase (G6PD) deficiency and had no other factors predisposing haemolysis. We reviewed the literature and discuss the relationship between glibenclamide and haemolytic crisis and between G6PD-deficiency and diabetes. PMID- 15126006 TI - Genetic polymorphism of methylenetetrahydrofolate reductase as a risk factor for diabetic nephropathy in Chinese type 2 diabetic patients. AB - OBJECTIVE: Genetic predisposition has been implicated in diabetic nephropathy (DN). The C677T variant of the methylenetetrahydrofolate reductase (MTHFR) gene, one of the key enzymes catalyzing remethylation of homocysteine, may play a role in the development of not only vascular disease but also diabetic microangiopathies. In this study, we examined the distribution of the MTHFR genotypes in the Chinese population and the association between the C677T variant and diabetic nephropathy. METHODS: 220 unrelated patients with type 2 diabetes mellitus and 130 controls were recruited. The MTHFR genotype was analyzed by PCR followed by HinfI digestion. Plasma total homocysteine levels were measured using high-performance liquid chromatography (HPLC) with fluorescence detection. RESULTS: In 130 healthy control subjects, the frequency of the mutant T allele was 30.0%, comparable to that of a Hong Kong (Chinese) population. The distribution of the three genotypes was as follows: TT genotype, 16.9%; CT genotype, 26.2%; and CC genotype, 56.9%. This genotype distribution did not differ between control subjects and type 2 diabetic patients in which 19.1% were TT, 34.5% were CT and 46.4% were CC (2=3.85, P>0.05). The frequency of the mutant T allele was 42.3% in diabetic patients with nephropathy (n=124) versus 28.6% in those without nephropathy (n=96). The genotype frequencies were TT, 21.0%; CT, 42.7%; CC, 36.3% in diabetic patients with nephropathy versus TT, 16.7%; CT, 23.9%; CC, 59.4% in those without nephropathy. The MTHFR genotype and allele frequencies were different between diabetic patients with and without nephropathy (chi2=12.27, P<0.005; chi2=8.77, P<0.005, respectively). Moreover, plasma homocysteine levels were markedly higher in individuals with TT genotype than those with CC or CT genotype. CONCLUSIONS: The C677T mutation of MTHFR gene is common in the Chinese population. MTHFR C677T gene polymorphism associated with a predisposition to increased plasma homocysteine levels may represent a genetic risk factor for diabetic nephropathy in Chinese type 2 diabetic patients. PMID- 15126007 TI - Mild renal insufficiency as a cardiovascular risk factor in non-proteinuric type II diabetes. AB - OBJECTIVES: To evaluate cardiovascular risk according to baseline renal function in a group of non-proteinuric type II diabetic patients. MATERIAL AND METHODS: Prospective study with a follow-up of 423 non-proteinuric type II diabetic patients with creatinine <150 micromol/l for an average of 4.7 years (S.D. 1.55). Creatinine clearance (CC) was estimated using the Cockcroft-Gault formula and expressed in millilitre per minute. The hazard ratio (HR) associated with each millilitre per minute decrease in baseline CC on fatal or non-fatal cardiovascular events and total mortality was evaluated using the Cox regression model. RESULTS: Baseline creatinine was 89 micromol/l (S.D. 15.9) and CC was 69.5 ml/min (S.D. 20). There were 63 cardiovascular events (15 unstable angina, 10 non fatal myocardial infarctions, 25 non-fatal strokes, two amputations, nine fatal myocardial infarctions and two fatal strokes) and 39 total deaths (11 for cardiovascular causes). The cardiovascular event rate was 31.7/1000 patient-years and the total mortality rate was 19.6/1000 patient-years. The independent predictors of cardiovascular events were: CC (HR=1.035; confidence interval (CI) 95% 1.02-1.05; P<0.0001), total cholesterol/HDL cholesterol ratio (HR=1.25; CI 95% 1.1-1.4; P=0.0008), baseline coronary heart disease (HR=2.05; CI 95% 1.07 3.9; P=0.04) and baseline microalbuminuria (HR=2.3; CI 95% 1.3-3.8; P=0.003). The independent total mortality predictors were: CC (HR=1.04; CI 95% 1.02-1.08; P<0.0001), male (HR=2.1; CI 95% 1.1-4; P=0.027) and baseline microalbuminuria (HR=2.1; CI 95% 1.1-4;P=0.03). CONCLUSIONS: Mild renal insufficiency increases cardiovascular risk in non-proteinuric patients with type II diabetes. PMID- 15126008 TI - Hypertensive disorders in Japanese women with gestational glucose intolerance. AB - We investigated the relationship between gestational glucose intolerance and the development of pregnancy-induced hypertension including gestational hypertension (GH) and preeclampsia. Consecutive Japanese women with singleton pregnancies underwent a standard 1h, 50g oral glucose challenge test (GCT) at 24-27 weeks of gestation, followed by a 75g, 2h oral glucose tolerance test (OGTT) if the GCT result exceeded 130mg/dl. Using criteria of the Japan Society of Obstetrics and Gynecology, gestational diabetes mellitus (GDM) was defined by two or more abnormal OGTT values and mild glucose intolerance by one abnormal value. The normal glucose tolerance group included women with GCT results below 130mg/dl or normal OGTT values. GH was defined as blood pressure of at least 140/90mmHg occurring for the first time after mid-pregnancy, without proteinuria. Preeclampsia was determined as GH with proteinuria. Of 2651 consecutive patients, 49 women were found to have GDM, and 139 showed mild glucose intolerance. Sixty patients showed GH, and 58 developed preeclampsia. The frequency of GH in mild glucose intolerance or GDM was 5.8% or 8.2%, respectively, significantly greater than in normal glucose tolerance (P<0.01). Incidence of preeclampsia was not significantly increased in women with mild glucose intolerance or GDM (2.2% or 4.1%, respectively, compared to those with normal glucose tolerance). Japanese women with gestational glucose intolerance therefore have an increased risk of developing GH. PMID- 15126009 TI - Gln27Glu polymorphism of the beta2 adrenergic receptor gene in healthy Japanese men is associated with the change of fructosamine level caused by exercise. AB - Adrenaline plays a major role in the maintenance of blood glucose level by promoting glycogenolysis during prolonged exercise predominantly via the beta2 adrenergic receptor (beta2AR). Because beta2ARs are mainly present in the muscle and liver, beta2AR gene polymorphism may affect changes in glucose metabolism caused by exercise. We, therefore, investigated the effect of beta2AR gene polymorphism on glucose metabolism in healthy Japanese men. The study group consisted of 124 unrelated healthy Japanese men who were aged 21-69 years (mean +/- S.D.: 45.3+/-11.7). They participated in an exercise program which was defined as low-moderate intensity at 20-60min per day, 2-3 days per week for 3 months. The genotype of Gln27Gln was detected in 109 subjects (87.9%), of Gln27Glu in 15 subjects (12.1%) and of Glu27Glu in none, and that of Arg16Arg, Arg16Gly and Gly16Gly in 32 (25.8%), 79 (63.7%) and 13 subjects (10.5%), respectively. There was no association between these polymorphisms and the metabolic characteristics at baseline. The change in fructosamine level as a result of exercise showed that the carrier of the Glu allele had a better response to exercise than the non-carrier. In conclusion, Gln27Glu polymorphism was associated with the change in fructosamine level resulting from exercise, but not Arg16Gly polymorphism. PMID- 15126010 TI - IGT with fasting hyperglycemia is more strongly associated with microalbuminuria than IGT without fasting hyperglycemia. AB - Previous studies have established that impaired glucose tolerance (IGT) patients with fasting hyperglycemia (IGT/FH: fasting plasma glucose (FPG) level 6.1-7.0 mmol/l and 2 h PG level of 7.8-11.1 mmol/l) exhibit higher insulin resistance than those with isolated IGT (FPG level <6.1 mmol/l and 2 h PG level of 7.8-11.1 mmol/l), but the association with microalbuminuria has not been determined. Here, we evaluate the prevalence of microalbuminuria in non-diabetic Japanese males 20 70 years of age. The subjects were classified into four groups based on the results of OGTT: normal glucose tolerance (NGT: n=71), impaired fasting glucose (IFG: n=24), isolated IGT (n=36), and IGT/FH (n=23). A urinary albumin-to creatinine ratio (ACR) from 30 to 300 microg/mg creatinine was counted as microalbuminuria. The prevalence of microalbuminuria was higher in subjects with IGT/FH than in subjects with isolated IGT (26% versus 14%). Logistic regression analysis showed microalbuminuria to be more significantly associated with IGT/FH (OR=3.82, 95% CI 1.09-13.36) than with isolated IGT (OR=1.75, 95% CI 0.50-6.17). While insulin resistance (HOMA-IR) in isolated IGT was not significantly different from that in NGT, insulin resistance in IGT/FH was significantly higher (P<0.01). Regression analysis of ACR in IGT showed a significant correlation with insulin resistance (P=0.012). Accordingly, microalbuminuria is more strongly associated with IGT/FH than with isolated IGT, most likely due to the higher insulin resistance. PMID- 15126011 TI - Retained capacity of glucose-mediated insulin secretion in patients with type 2 diabetes mellitus inversely correlates with the duration of diabetes. PMID- 15126012 TI - Metformin attenuates progression of carotid arterial wall thickness in patients with type 2 diabetes. AB - To investigate the anti-atherogenic effect of metformin, we prospectively evaluated the effect of metformin treatment on common carotid intima-media thickness (CCA-IMT) in patients with type 2 diabetes. A 2-year open prospective study was performed. Thirty-six patients were treated with metformin (500-750 mg per day). CCA-IMT was measured after 1- and 2-year treatment. Changes in CCA-IMT were compared with control patients. After 2-year metformin therapy, the progression of CCA-IMT was significantly less than 56 control patients (0.02+/ 0.08 mm versus 0.07+/-0.08 mm, P<0.01). Metformin therapy did not alter body weight, blood pressure, HbA1c, and serum lipids relative to the control. Thus, metformin attenuates the progression of CCA-IMT. This anti-atherogenic effect is not mediated through changes in classical cardiovascular risk factors. PMID- 15126013 TI - Type-1 diabetes mellitus with insufficient serum immunoreactive insulin elevation after subcutaneous NPH-insulin injection. PMID- 15126014 TI - Pediatric myasthenia gravis and velopharyngeal incompetence. AB - OBJECTIVE: To determine the clinical course of velopharyngeal incompetence in children with myasthenia gravis (MG). METHODS: A 30-year retrospective study was performed using the medical records of 538 children who presented with velopharyngeal inadequacy (VPI) to a tertiary care academic pediatric center. Children with velopharyngeal incompetence due to myasthenia gravis were identified and their clinical courses were reviewed. RESULTS: Four children were identified with velopharyngeal incompetence associated with myasthenia gravis. All four children required intervention for improvement of speech intelligibility. A speech prosthesis was the uniform intervention. CONCLUSION: Neonatal myasthenia gravis patients should be followed long-term as symptoms may recur as speech impairment. In addition, a high index of suspicion for this entity is required for early diagnosis due to the highly variable presentation and clinical course. PMID- 15126015 TI - Tympanometry measures in native and non-native Hawaiian children. AB - BACKGROUND: Ethnicity has been readily accepted as a variable affecting the incidence of otitis media, with certain indigenous groups having an increased risk of middle ear dysfunction. Tympanometry provides objective information on middle ear status, and findings obtained from this procedure have often served as a criterion for medical referral. OBJECTIVE: To extend previous research and to facilitate use of normative tympanometry measures obtained from children with native Hawaiian ancestry. METHODS: Data were collected from 718 ears of 359 children in academic levels ranging from preschool to third grade. Subjects were matched across groups (182 native Hawaiian; 177 non-native Hawaiian) for academic level and gender. Variables included physical ear-canal volume (Vec), tympanometric peak compliance (peak Y, also known as static admittance), tympanometric width (TW), and tympanometric peak pressure (TPP). RESULTS: Significantly higher TW (F1,714=8.82, P=0.008) and TPP (F1,714=9.98, P=0.002) values occurred in ears of native Hawaiian children. Statistical interaction between gender and age was not significant. CONCLUSION: Differences in tympanometric findings between groups suggest differences in middle ear function, and these findings continue to underscore the importance of including tympanometry within a hearing screening protocol for early identification of possible hearing impairment. PMID- 15126016 TI - Bacteriology of medically refractory acute otitis media in children: a 9-year retrospective study. AB - OBJECTIVE: To identify the causative organisms for medically refractory acute otitis media (AOM) in children, and to recommend the appropriate antibiotics for these patients. STUDY DESIGN: Retrospective chart review. METHODS: The medical records for 671 children (1258 ears) undergoing myringotomies between January 1993 and December 2001 were retrospectively reviewed. "Medically refractory" AOM is defined as AOM requiring emergency myringotomy because of toxicity persisting despite second-phase antibiotics. Of these, 18 children were enrolled in our study, with 25 of the ears affected by medically refractory AOM. Myringotomy was performed for pus drainage, bacterial culture and susceptibility studies in all cases. Clinical factors were analyzed to demonstrate possible relationships with microbiological findings. RESULTS: Bacteria were recovered in specimens taken from 12 ears. The positive culture rate was 48%. Bacteriological studies showed mostly growing gram-positive bacteria, such as coagulase negative staphylococcus, Staphylococcus aureus and Streptococcus pneumoniae (in decreasing order of incidence). The sensitivity rate to pre-myringotomy antibiotics was 85.7%. With the exception of Pseudomonas aeruginosa and two ears with intracranial complications or pneumonia, all other isolated microorganisms were susceptible to first-generation cephalosporins. No statistically significant differences were noted between culture rates and clinical factors. CONCLUSION: The distribution of organisms in our medically refractory AOM cases differed from that for simple AOM, with gram-positive bacteria having significantly higher rates of incidence. Combining intravenous aminoglycoside with first-generation cephalosporin is considered an inexpensive and effective treatment covering all potential microorganisms. Vancomycin or third-generation cephalosporins are only needed for major complications or other coincident infections extant during the initial presentation. PMID- 15126017 TI - Salivation in healthy schoolchildren. AB - OBJECTIVE: To obtain reference data on the unstimulated salivary flow rate in healthy 6-11-year-old boys and girls, for use in the treatment of drooling in mentally and physically handicapped children. METHODS: Under standard conditions unstimulated saliva was collected from 62 healthy children using the swab method. Flow rate was determined by calculating the increase in weight of cotton rolls placed in the mouth for 5 min. The influence of the variables age, gender and type of gland were evaluated. RESULTS: No significant difference was found between the flow rate in boys and girls or between the children in this age window. The swab method showed good reproducibility. The right and left parotid glands had equal flow rates of 0.15 ml/min. The flow rate in the floor of the mouth secreted by the submandibular and sublingual glands was 0.32 ml/min. CONCLUSIONS: In healthy children of 6-11 years, age and gender did not influence the salivary flow rate and therefore do not have to be taken in consideration when measuring salivary flow rate. The swab method was reliable, easy to carry out and well-tolerated by the group of schoolchildren. PMID- 15126018 TI - The relationship of tonsillar hyperplasia and asthma in a group of asthmatic children. AB - BACKGROUND: The decline of infections in childhood may contribute to the rising severity and prevalence of atopic disorders in developed countries. With this regard, we examined the relationship of frequent tonsillitis and consequent tonsillar hyperplasia with the development of asthma. METHODS: Sixty-seven asthmatic children (ages 3-14) who had no signs or symptoms of acute tonsillitis were included. The control group consisted of 92 randomly selected children who had no signs or symptoms of asthma or acute tonsillitis. Parents were interviewed about the incidence of tonsillitis diagnosed by physicians and history of tonsillectomy; tonsil sizes were evaluated by oropharyngeal inspection by the same observer using the Brodsky L. Scala. RESULTS: A statistically significant association is found between frequent tonsillitis and consequent tonsillar hyperplasia with the development of asthma. CONCLUSIONS: Our data suggests that recurrent tonsillitis is associated with a decline in the prevalence of asthma by inducing a Th 1 predominant immune response. Our findings are compatible with the hygiene hypothesis. PMID- 15126019 TI - Current practice patterns in tonsillectomy and perioperative care. AB - OBJECTIVES: Tonsillectomy is one of the most commonly performed otolaryngologic procedures in the United States. Many options and controversies exist regarding techniques and peri-operative management. The purpose of the study was to examine current practice patterns among otolaryngologists regarding tonsillectomy. METHODS: A 13 question survey regarding tonsillectomy techniques and peri operative management was mailed to 10% of randomly selected board certified otolaryngologists of the AAO-HNS in the spring of 2002. Four hundred and eighteen anonymously completed questionnaires were returned, for a response rate of 58.5%. Statistical analysis of survey data was performed by means of cross tabulation and Pearson Chi-Square Calculation. RESULTS: Monopolar electrocautery was the most common technique used among those surveyed (53.5%). There was a significant correlation between choice of monopolar electrocautery and the cited reason for choice of technique being decreased blood loss (P < 0.001). There was no relationship between pediatric fellowship training and choice of technique. 97.7% routinely admitted sleep apnea patients for post-operative observation. There was no significant correlation between practice setting (tertiary versus community) and type of post-operative monitoring for sleep apnea patients, with patients most commonly admitted to an intermediate care setting. CONCLUSION: In our survey, the most common surgical technique for tonsillectomy was monopolar electrocautery, chosen for the reason of decreased blood loss. PMID- 15126020 TI - Nutritional supplements as adjunctive therapy for children with chronic/recurrent sinusitis: pilot research. AB - OBJECTIVE: Inflammation and edema of the sinonasal mucosa are important in the pathophysiology of sinusitis. Based on the similarities between otitis media (OM) and sinusitis, and our previous research on OM, we hypothesized that nutritional supplements would be effective adjunctive therapy for the treatment of children with chronic/recurrent sinusitis. METHODS: We performed a 4 month, open-label, dose-titration study; subjects were enrolled from late January to early March 2003. Each subject served as his own control. Study supplements were a lemon flavored cod liver oil and a children's multivitamin-mineral with selenium, prescribed in escalating doses; at higher doses, fish oil was substituted for cod liver oil. Subjects were private pediatric otolaryngology outpatients with a clinical diagnosis of chronic/recurrent sinusitis, whose symptoms were refractory to treatment with antibiotics. RESULTS: Our four subjects were Caucasian males, ranging in age from 4.2 to 9.8 years, with chronic/recurrent sinusitis for at least 3 years prior to entry in the study. Three subjects had a positive response; one subject dropped out for administrative reasons. Four, six, and eight weeks after beginning study supplements, the responders had decreased sinus symptoms, fewer episodes of acute sinusitis, and fewer doctor visits for acute illnesses. Their parents reported that they had begun to recover from upper respiratory illnesses without complications, which was unusual for these children, as was improvement in springtime; their improvement had previously been limited to the summer months or periods of home-schooling. CONCLUSIONS: Use of flavored cod liver oil and a multivitamin-mineral with selenium as adjunctive therapy for children with chronic/recurrent sinusitis is an inexpensive, non invasive intervention that clinicians can use for selected patients, pending the performance of definitive, large, well-controlled studies. PMID- 15126021 TI - Racial/ethnic disparities in the diagnosis of otitis media in infancy. AB - OBJECTIVE: Otitis media (OM) is an extremely common pediatric diagnosis. Several risk factors have been associated with OM, but the relationship between OM and race/ethnicity remains controversial. We sought to define the relationship between OM diagnosis and race/ethnicity in infants. METHODS: By multivariable logistic regression, we evaluated the association between OM diagnosis and race/ethnicity in 11,349 non-low-birthweight infants who were participants in a prospective cohort study of infant care practices. RESULTS: As in previous studies, breastfeeding was associated with a decreased risk of OM diagnosis while other factors were independently associated with a substantially increased risk of OM diagnosis: out-of-home daycare, multiple children living in the home, and mother's multiparity. Daycare was associated with a "dose effect" in that the risk of OM diagnosis increased with an increasing number of children in the daycare. While the crude analysis suggested little relation of OM diagnosis and race/ethnicity, the association was confounded by several covariates including maternal marital status, number of children living in the home, breastfeeding status, and maternal age. After adjustment for relevant confounders, Black (OR 0.74; 95% CI 0.61-0.89) and Asian infants (OR 0.77; 95% CI 0.57-1.0]) were less likely to be diagnosed with OM than White infants. CONCLUSIONS: This large prospective study confirms previous risk factors for OM and demonstrates a strong "dose effect" of the size of daycare centers on OM. The study also demonstrates that the association between race/ethnicity and OM diagnosis is confounded by social factors. After adjusting for such factors, Black and Asian infants are less likely to be diagnosed with OM than White infants. The reason for this racial disparity remains unknown. PMID- 15126022 TI - Universal newborn hearing screening program in Bulgaria. AB - The design of a Universal Newborn Hearing Screening Program (UNHSP) depends on many factors unique to each hospital. The goal of any UNHSP is to perform a valid hearing screening on all newborn infants prior to hospital discharge. Ultimately hearing screening of all infants should decrease the age at which intervention is initiated in these who require it. It is clear that there is no "one" model for UNHSPs. Each program must carefully consider what type and severity of hearing loss it wishes to identify. Then, based on available screening tools, program philosophy, prior experience, personnel, typical maternity length of stay, etc., an appropriate protocol must be developed. We present our protocol for detection and confirmation of hearing loss, and for habilitation of deaf children. For detection the hearing loss we used the automated auditory brainstem response (ABR) delivery system (A-ABR), and we confirm this with conventional non automated ABR delivery system (BERA). The healthy babies we screen after 6h of age using a simultaneous 35-dB nHL screening option available on the Newborn Hearing Screener ALGO-2. The neonatal intensive care unit (NICU) babies we screen prior to discharge using all options of the ALGO-2. The "refer" ALGO-2 results are immediately retested. Infants in need of an outpatient screen are recalled at age 3-4 weeks. The diagnosis "hearing loss" is determinate only by BERA and the neonatal hearing screening is the first part of program of habilitation of hearing-impaired children. The cost for UNHSP in Bulgaria is 1407 euro per case identified. PMID- 15126023 TI - The effects of incisional myringotomy and CO2 laser myringotomy on rat tympanic membranes. AB - OBJECTIVE: The purpose of this study was to compare closure rates and histopathological findings of carbon dioxide (CO(2)) laser myringotomies to those of incisional myringotomy. STUDY DESIGN: We performed CO(2) laser round myringotomy on left ears and incisional round myringotomy on the right ears of 34 rats on the same day. The incisions were 2mm in size on both ears. Examination was made with otoendoscope on days 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 25, 30, 35, 40, 45, 50 with videorecording. Two rats were sacrificed every 2 days for 25 days and every 5 days thereafter, randomly. The tympanic membranes (TM) were excised and hyalinization, fibrosis and inflammation were assessed with light microscope. RESULTS: Most of the CO(2) laser myringotomies healed after day 15, and all of them healed by day 50. Three of the incisional myringotomy perforations were closed at day 3, and the rest by day 15. Patency of CO(2) laser myringotomies was significantly longer than that of incisional myringotomies. Hyalinization, fibrosis and inflammation of the incisional myringotomy group were significantly less than those of the laser myringotomy group. CONCLUSIONS: CO(2) laser myringotomies remain patent for a longer period of time than the incisional procedure, however, they cause more tissue inflammation. We believe that CO(2) laser myringotomy is an effective method, however, additional studies are needed to identify its complications. PMID- 15126024 TI - Behcet's disease: external ear involvement. AB - Behcet's disease is an autoimmune vasculitis characterized by recurrent episodes of oral and genital ulcerations, skin lesions and uveitis. It also affects neurological, vascular, articular and gastrointestinal systems. Although etiology and pathogenesis are under investigation, studies have shown that HLA-B51 influences genetic susceptibility. The development of the disease is associated to neutrophil activation. Treatment depends on the severity of the disease and immunosuppressive drugs are often used. Blindness and dementia are long-term concerns. We report an unprecedented case of a child with Behcet's disease presenting a skin lesion with necrosis of the external ear canal, facial paresis, Horner's syndrome and rupture of the internal carotid artery. PMID- 15126025 TI - Congenital midnasal stenosis in an infant. AB - Neonates and infants are obligate nasal breathers and nasal obstruction in this age group can be a life threatening emergency. Even though the commonest cause for nasal obstruction is nasal oedema, bony stenosis of the posterior choanae or pyriform aperture is quite common and may be seen on CT scans. We describe a case of mid-nasal stenosis in a neonate and discuss the aetiology and management of nasal obstruction in this age group. PMID- 15126026 TI - Foregut duplication cyst arising in the floor of mouth. AB - Enteric duplications are uncommon. A rare foregut duplication cyst arising in the floor of mouth is reported. Diagnosis of cystic lesion within the oral cavity was made in utero. Enteric duplications have not yet been attributed to a single embryogenesis but are likely to represent an error in migration of normal cells that rest in an abnormal position. Immediately following delivery of the neonate via an ex-utero intrapartum treatment (EXIT) procedure, the cyst was marsupialised but required definitive surgical excision at 5 weeks of age. PMID- 15126027 TI - Esophageal perforation preceding fatal closed head injury in a child abuse case. AB - This is a case describing an extensive esophageal perforation in a 15-day-old infant with an unclear mechanism of injury, who at 3 months of age presented as the victim of child abuse from a fatal closed head injury. Extensive esophageal perforation in a neonate or infant is usually induced by trauma, either accidental or non-accidental. When the given history does not correlate with the degree of injury, child abuse must be strongly suspected and a thorough social investigation must be undertaken. PMID- 15126028 TI - Intramural foreign body in oesophagus. AB - Intramural foreign body in oesophagus is very rare and is mostly associated with complications leading to high morbidity and mortality. Recently we came across a coiled metallic wire discovered unexpectedly in a chest X-ray of a 6-month-old male baby. This was further investigated and was found to be located intramurally in the oesophagus. It was successfully removed with rigid oesophagoscopy performed under telescopic guidance. No such foreign body has been reported in an infant so far in English literature. PMID- 15126029 TI - Sudden hearing loss due to large vestibular aqueduct syndrome in a child: should exploratory tympanotomy be performed? AB - A 16-year-old girl applied to our ENT clinic with a 3-day history of right hearing loss, tinnitus, and pressure in the right ear. She had had surgery for right perilymph fistula two times, one at the age of 7 and the second at the age of 9. She had recovered after both of these surgeries. This time she had exploratory tympanotomy and perylymh fistula was detected. Computerized tomography investigation obtained after 5 days postoperatively showed bilateral large vestibular aqueducts and otherwise normal inner ear structures. Thyroid function tests and neck palpation were normal. It was an unusual case with both large vestibular aqueduct syndrome (LVAS) and simultaneous spontaneous perilymph fistula. PMID- 15126030 TI - Somatostatin treatment of massive lymphorrhea following excision of a lymphatic malformation. AB - Postoperative lymphorrhea is a serious and occasionally lethal complication of neck dissection and cardiothoracic surgery. Management is not standardized, but usually centers around diet modification, drainage, pressure dressings, and reoperation. We report the successful use of the long acting somatostatin analogue octreotide in the management of massive lymphorrhea complicating excision of a large cervicomediastinal lymphatic malformation in an infant. Based on this report and a review of the available literature, we advocate the early consideration of somatostatin and its analogues in the control of lymphorrhea. PMID- 15126031 TI - Traumatic supraglottitis. AB - Traumatic supraglottitis may occur after mechanical, thermal, chemical, or other airway injury. It can be managed following the principles used for managing the airways of patients with infectious supraglottitis. A protocol for evaluation and management is described. PMID- 15126032 TI - Comparison of neuroplastin and synaptic marker protein expression in acute and cultured organotypic hippocampal slices from rat. AB - Organotypic hippocampal slice cultures can be used to study hippocampal biochemistry and physiology over a chronic period on the days to weeks timescale. In order to validate the organotypic hippocampal slice culture for our ongoing studies of synaptic function, we have compared, using Western blotting, the levels of a number of synaptic proteins from in vitro organotypic hippocampal slice cultures with those from in vivo hippocampal slices prepared from age matched controls. We chose to follow the developmental expression of the neuroplastin (np) family of immunoglobulin related cell adhesion molecules (CAMs), np65, a brain specific isoform highly expressed in hippocampal neurones and np55 a more widely expressed isoform and two synaptic marker proteins, synaptophysin, a pre-synaptic marker and post-synaptic density protein-95, PSD95, a post-synaptic marker. All showed increasing expression over the developmental time period, both in vivo and in vitro. The level of both neuroplastins was also consistent between the in vivo and in vitro preparations, whereas the level of PSD95 was markedly increased in the organotypic hippocampal slice cultures while the level of synaptophysin was slightly decreased. Whilst these findings may indicate some differences in the composition and organisation of synapses, the developmental expression profiles of these synaptic proteins within organotypic hippocampal slice cultures suggests they are a valid model for the study of synapse function and development in vitro. PMID- 15126033 TI - Effects of prenatal protein malnutrition and acute postnatal stress on granule cell genesis in the fascia dentata of neonatal and juvenile rats. AB - Although postnatal genesis of granule cells in the hippocampal fascia dentata is known to be influenced by prenatal protein deprivation or by stress, the combined effects of prenatal protein malnutrition and stress on these cells are unknown. This study was designed to examine this combined effect. Well-nourished and prenatally malnourished pups on postnatal day 7 (P7) were stressed by maternal separation and reduction of body temperature and on postnatal day 30 (P30) by immobilization with restraint. Bromodeoxyuridine (BrDU) was injected at the time of stress, and 2 h later, the numbers of immunolabeled cells were quantified by standard stereological techniques. In comparison to controls, prenatally malnourished rats showed a significantly lower number of cells tagged in the fascia dentata on P7 (p < or =0.05), and a significantly higher number of cells (p < or =0.05) on P30. In both age groups, control rats exposed to acute stress showed a significantly decreased number of cells tagged in the fascia dentata (p < or =0.05). In contrast, neurogenesis in malnourished rats was not significantly affected by acute stress at either age. Thus, the pattern of neurogenesis in the fascia dentata and its response to stress has been fundamentally altered by prenatal protein deprivation. PMID- 15126034 TI - The mGlu5 metabotropic glutamate receptor is expressed in zones of active neurogenesis of the embryonic and postnatal brain. AB - Metabotropic glutamate (mGlu) receptors have been implicated in the regulation of developmental plasticity. Here, we examined the expression of mGlu1a-b, -2, -3, 4a-b, and -5a receptor subtypes from embryonic day 12 (E12) to the early and late postnatal life. While all transcripts (with the exception of mGlu4 mRNA) were detected prenatally, only the mGlu5 receptor protein was found in detectable amounts in the embryonic brain. Immunohistochemical analysis showed that the mGlu5 receptor was mainly expressed by cells surrounding the ventricles at E15, whereas it was more diffusely expressed at E18. In the postnatal life, besides its classical expression sites, the mGlu5 receptor was found in zones of active neurogenesis such as the external granular layer (EGL) of the cerebellar cortex and the subventricular zone. In these regions, the presence of actively proliferating progenitor cells was detected by BrdU staining. No other subtype (among those we have examined) was found to be expressed in regions enriched of BrdU(+) cells. These data suggest a role for mGlu5 receptors in the early brain development and in basic cellular processes such as proliferation and/or differentiation. PMID- 15126035 TI - Support of trigeminal sensory neurons by nonneuronal p75 neurotrophin receptors. AB - The p75 neurotrophin receptor (p75NTR) binds all four mammalian neurotrophins, including neurotrophin-3 (NT-3) required for the development of select sensory neurons. This study demonstrated that many gustatory and somatosensory neurons of the tongue depend upon p75NTR. Each of thousands of filiform papillae at the front of the tongue as well as each somatosensory prominence at the back of the tongue has a small cluster of p75NTR-positive epithelial cells that is targeted by somatosensory innervation. This expression of p75NTR by epithelial target cells required NT-3 but not adult innervation. NT-3-secreting cells were adjacent to the p75NTR-positive target cells of each somatosensory organ, as demonstrated in NT-3(lacZneo) transgenic mice. In NT-3 null mutant mice, there were few lingual somatosensory neurons. In p75NTR null mutant mice, the lingual somatosensory axons were likewise absent or had deficient terminal arborizations. Cell culture indicated that substrate p75NTR can influence neuronal outgrowth. Specifically, dissociated trigeminal sensory neurons more than doubled their neurite lengths when grown on a lawn of p75NTR-overexpressing fibroblasts. This enhancement of neurite outgrowth by fibroblast p75NTR raises the possibility that epithelial target cell p75NTR may help to promote axonal arborization in vivo. The co-occurrence in p75NTR null mice of a 35% reduction in geniculate ganglion taste neurons and a shortfall of taste buds is consistent with the established role of gustatory innervation in prompting mammalian taste receptor cell differentiation. PMID- 15126036 TI - Effect of Brn-3a deficiency on parvalbumin-immunoreactive primary sensory neurons in the dorsal root ganglion. AB - Immunohistochemistry for parvalbumin, a marker for primary proprioceptors, was performed on the dorsal root ganglion (DRG) of wildtype and knockout mice for Brn 3a at postnatal day 0 and embryonic day 18.5. The DRG contained many parvalbumin immunoreactive (ir) neurons in wildtype (5.4%) and knockout mice (5.6%). Cell size analysis demonstrated that such neurons were mostly medium-sized to large in these mice. Therefore, it is unlikely that the survival of proprioceptors is dependent upon Brn-3a in the DRG. In the dorsal column and gray matter of the spinal cord of knockout mice, however, parvalbumin-ir nerve fibers were sparse compared to wildtype mice. The number of parvalbumin-ir varicosities around motoneurons decreased in the mutant. Thus, our data suggest that Brn-3a may play an important role in the central projection and terminal formation of DRG proprioceptors in the spinal cord. PMID- 15126037 TI - Postnatal development of the afferent projections from the dorsomedial thalamic nucleus to the frontal cortex in mice. AB - The postnatal development of mediodorsal thalamic projections to the dorsomedial frontal cortex of mice was assessed by means of the retrograde peroxidase colloidal gold complex tract tracing system. The tracer was injected into the dorsomedial frontal cortex from the day of birth (P0) to 60 days of postnatal age (P60). Since birth, a dense retrograde labeling has been found in the mediodorsal nucleus, which increased progressively from P4 to P8 and began to decrease at P10 until P13 (67.37% vs. the maximal average, P4). After P16, the mean average remains stable up to P60. PMID- 15126038 TI - Development of AT(1) and AT(2) receptors in the ovine fetal brain. AB - This study determined the development of AT(1) and AT(2) receptors in the ovine fetal brain from preterm to term by utilizing Western blot for the receptor expression at the protein level, RT-PCR for the receptor mRNA, and immunostaining for the specific receptor immunoreactivity. The results demonstrated that AT(1) and AT(2) receptors developed in an increasing pattern from preterm to term gestational periods in the fetal sheep brain. Both AT(1) and AT(2) receptors have appeared in the major structures in the angiotensin-related central cardiovascular and body fluid controlling pathways at the 0.7 of the gestational age. Importantly, AT(1) receptors have been discovered in the supraoptic nuclei in the fetal hypothalamus, and in the lateral parabrachial nuclei and the ventrolateral medulla in the fetal hindbrain. This provides evidence of the anatomical existence of the angiotensin receptors in the brain areas that are critical for cardiovascular and fluid regulatory functions in utero. In addition, although the results demonstrated the predominance of AT(2) receptors in several regions such as the cerebellum in the ovine fetal brain, dominant occupation of AT(1) receptors in the hypothalamus have appeared early in the life of sheep animals before birth. Together, the data support the hypothesis that the central angiotensin receptors are well developed and established in the last third trimester of gestation. The brain receptors provide a pharmacological basis for the action of angiotensin in the maintenance of in utero fetal physiological functions, including cardiovascular and body fluid balance. PMID- 15126039 TI - Cadherin-2 and cadherin-4 in developing, adult and regenerating zebrafish cerebellum. AB - Cadherins are cell adhesion molecules that regulate development of a variety of tissues and maintenance of adult structures. In this study, we examined expression of two zebrafish classical cadherins, cadherin-2 and cadherin-4, in the cerebellum of developing, normal adult, and regenerating adult zebrafish using in situ hybridization and immunohistochemical methods. Cadherin-2 was widely expressed by the cerebellum of embryonic (24-50-h post fertilization) and larval zebrafish (3-14 days). Cadherin-2 expression became much reduced in the adult cerebellum, but it was greatly up-regulated in the regenerating adult cerebellum. Cadherin-4 was not detected in the embryonic cerebellum, but it was expressed in the Purkinje cells of the larval and adult cerebellum. To gain insight into cadherin-2 role in the formation of the cerebellum, we analyzed embryos injected with a specific cadherin-2 antisense morpholino oligonucleotide (cdh2MO1), and found that the cerebellar development of the cdh2MO1-injected embryos was severely disrupted. This phenotype was confirmed by examining a cadherin-2 mutant, glass onion. Our results suggest that cadherins are crucial for the normal development of the zebrafish cerebellum, and they may also be involved in the regeneration of injured fish cerebellum. PMID- 15126040 TI - Axonal regeneration of proctolinergic neurons in the central nervous system of the locust. AB - We provide evidence for axonal regeneration in the central nervous system (CNS) of the locust (Locusta migratoria). We followed the morphology of a small set of proctolin-immunoreactive neurons in the ventral nerve cord before and after crushing one cervical connective in the third instar. The proximal segments started sprouting within 3 days post lesion and grew into the suboesophageal ganglion within 9 days, covering a distance of approximately 2 mm. Within the suboesophageal ganglion, the regenerated neurites formed arborisations in the appropriate region which closely resemble the original shape. These findings will allow us to compare regeneration to the well-described embryonic development of axonal connectivity in this animal. PMID- 15126041 TI - Optical imaging of intrinsic signals: recent developments in the methodology and its applications. AB - Since optical imaging (OI) of intrinsic signals was first developed in the 1980s, significant advances have been made regarding our understanding of the origins of the recorded signals. The technique has been refined and the range of its applications has been broadened considerably. Here we review recent developments in methodology and data analysis as well as the latest findings on how intrinsic signals are related to metabolic cost and electrophysiological activity in the brain. We give an overview of what optical imaging has contributed to our knowledge of the functional architecture of sensory cortices, their development and plasticity. Finally, we discuss the utility of OI for functional studies of the human brain as well as in animal models of neuropathology. PMID- 15126042 TI - A computational tool to simulate correlated activity in neural circuits. AB - A new computational approach to study correlated neural activity is presented. Simulating Elementary Neural NEtworks for Correlation Analysis (SENNECA) is a specific-purpose simulator oriented to small circuits of realistic neurons. The model neuron that it implements can reproduce a wide scope of integrate-and-fire models by simply adjusting the parameter set. Three different distributions of SENNECA are available: an easy-to-use web-based version, a Matlab (Windows and Linux) script, and a C++ class library for low-level coding. The main features of the simulator are explained, and several examples of neural activity analysis are given to illustrate the potential of this new tool. PMID- 15126043 TI - Histopathological and behavioral characterization of a novel model of cardiac arrest and cardiopulmonary resuscitation in mice. AB - Cardiac arrest is associated with high mortality and poor neurological outcome. We characterized functional and histological outcome in a novel mouse model of cardiac arrest and cardiopulmonary resuscitation (CPR) in order to study neuroprotective mechanisms. Cardiac arrest was induced in male C57Bl/6 and 129SVEV mice by i.v. injection of KCl. After 10 min cardiac standstill, CPR was initiated by administration of epinephrine, ventilation with 100% oxygen and chest compressions. Twenty-four hours before and 3 or 7 days after CPR, mice were subjected to behavioral testing using a passive avoidance task, locomotor activity in an open field, and spontaneous alternation in a T-maze. Hippocampal and caudoputamen injury was quantified 3 or 7 days after CPR. At both time points, caudoputamen injury was worse in 129SVEV mice. Post-ischemic mice of both strains showed a reduced number of correct choices in the T-maze up to 7 days after CPR, and were temporarily impaired in learning the passive avoidance task with a retention deficit on day 3 but not on day 7. Locomotor activity showed strain differences with C57Bl/6 mice being more active, but little ischemia related effects. A dissociation between functional and histological outcome was found emphasizing the importance of combining both outcome measures for evaluation of neuroprotective strategies. PMID- 15126044 TI - The end-to-side peripheral nerve repair. Functional and morphometric study using the peroneal nerve of rats. AB - Morphologic and functional recovery following an end-to-side repair was studied comparatively with conventional end-to-end repair in a model of peroneal nerve lesion in rats. Twenty-eight rats were used and divided into four groups according to the reparative procedure following nerve division: (1) nerve stumps buried into neighboring muscles (n = 8); (2) conventional end-to-end repair (n = 7); (3) end-to-side repair onto the tibial nerve (n = 8); (4) sham operation (n = 5). The sciatic functional index (SFI) was evaluated at weekly intervals for 8 weeks, the peroneal nerve being resected on the 56th day for histologic and morphometric studies. The SFI progressively improved in Groups 2 (-16.9) and 3 ( 22.7), although it did not reach normal values (around -8). The average nerve fiber density increased to normal values in both Groups 2 and 3, although accompanied by a marked decrease of average minimal and maximal nerve fiber diameter, myelin sheath area and G quotient. The differences between Groups 2 and 3 or Groups 2 and 4 were not significant. We conclude that, although resulting in significant morphologic and functional recovery, end-to-side repair is not as efficient as the conventional end-to-end nerve repair. However, end-to-side repair has a potential for application in selected cases in humans. PMID- 15126045 TI - Multi-electrode recording from the developing visual pathway of awake behaving ferrets. AB - We have developed an effective technique for obtaining multi-electrode extracellular recordings from developing lateral geniculate nucleus (LGN) and visual cortex in awake behaving ferrets. Using this approach, we have been studying changes in the spatio-temporal patterns of spontaneous activity within the visual pathway during pre eye-opening development. In this paper, we describe the fabrication and implantation of 8- and 16-channel drivable multi-electrode arrays, which enable us to obtain high-quality multi-unit recordings from nearly 100% of our recording sites. PMID- 15126046 TI - Do alternate methods of analysing motor evoked potentials give comparable results? AB - This study assessed the reliability of alternate methods of analysis of motor evoked potentials (MEPs) elicited by transcranial magnetic stimulation (TMS). We recorded two sets of MEPs (Time 1 and Time 2) at the optimal scalp sites for both the right first dorsal interosseous (FDI) and flexor carpi ulnaris (FCU) at two different stimulation intensities in 10 healthy subjects. MEP magnitude was determined in each of the following three ways: the mean peak-to-peak amplitude and area of the 20 individual responses; the amplitude and area of the ensemble averaged waveform; and the amplitude and area of the maximal response. There was no significant difference in amplitude or area for either muscle using any of the three methods between Time 1 and 2. However, the ensemble average (area and amplitude) was significantly smaller that the mean MEP, and the maximal MEP amplitude was significantly larger. Intraclass correlation analysis demonstrated that reliability of MEP measures over time was poor regardless of method. Reliability was similar between methods for FDI, but FCU had lower reliability values for the mean and ensemble average methods than the maximal method. PMID- 15126047 TI - Single extraction protocol for the analysis of 8-hydroxy-2'-deoxyguanosine (oxo8dG) and the associated activity of 8-oxoguanine DNA glycosylase. AB - Determination of the promutagenic base 8-hydroxy-2'-deoxyguanosine (oxo(8)dG) has been the hallmark of studies aimed to determine oxidative damage to DNA. Different techniques including HPLC, GC-mass spectrometry, DNA sensitive sites and histology have been used to quantify oxo(8)dG levels in samples from different sources. The most accepted and well-established methods are based on HPLC and the ability of oxo(8)dG to be oxidized with an electrochemical detector. Considerable concerns have been raised in the ability of different labs to utilize a process of DNA extraction that reduces the levels of artifactual oxo(8)dG formed during sample workup. Here, we present a fully detailed protocol that has been extensively used in our Lab to extract and analyze DNA and has little or no impact in the basal levels of oxo(8)dG. Additionally, this protocol allows for the determination of the activity of mOgg1, the enzyme responsible for the initial step in the repair of the accumulated oxo(8)dG, in the same sample in which oxo(8)dG is detected. PMID- 15126048 TI - Multivariate receptive field mapping in marmoset auditory cortex. AB - We describe a novel method for estimation of multivariate neuronal receptive fields that is based on least-squares (LS) regression. The method is shown to account for the relationship between the spike train of a given neuron, the activity of other neurons that are recorded simultaneously, and a variety of time varying features of acoustic stimuli, e.g. spectral content, amplitude, and sound source direction. Vocalization-evoked neuronal responses from the marmoset auditory cortex are used to illustrate the method. Optimal predictions of single unit activity were obtained by using the recent-time history of the target neuron and the concurrent activity of other simultaneously recorded neurons (R: 0.82 +/- 0.01, approximately 67% of variance). Predictions based on ensemble activity alone (R: 0.63 +/- 0.18) were equivalent to those based on the combination of ensemble activity and spectral features of the vocal calls (R: 0.61 +/- 0.24). This result suggests that all information derived from the spectrogram is embodied in ensemble activity and that there is a high level of redundancy in the marmoset auditory cortex. We also illustrate that the method allows for quantification of relative and shared contributions of each variable (spike train, spectral feature) to predictions of neuronal activity and describe a novel "neurolet" transform that arises from the method and that may serve as a tool for computationally efficient processing of natural sounds. PMID- 15126049 TI - Optimization of multiplexed bead-based cytokine immunoassays for rat serum and brain tissue. AB - The ability to simultaneously quantify multiple signaling molecule protein levels from microscopic neural tissue samples would be of great benefit to deciphering how they affect brain function. This follows from evidence that indicates signaling molecules can be pleiotropic and can have complex interactive behavior that is regionally and cellularly heterogeneous. Multiplexed examination of tissue proteins has been exceedingly difficult because of the absence of available techniques. This void now has been removed by the commercial availability of bead-based immunoassays for targeted proteins that allow analyses of up to 100 (6-150 kDa) proteins from as little as 12 microl. Thus far used only for sera (human and mouse) and culture media, we demonstrate here that sensitive (as low as 2 pg/ml), wide-ranging (up to 2-32 000 pg/ml), accurate (8% intra assay covariance) and reliable (4-7% inter-assay covariance) measurements can be made of nine exemplary cytokines (e.g., IL-1alpha, IL-1beta, IL-2, IL-4, IL-6, IL 10, GM-CSF, IFN-gamma, TNF-alpha) simultaneously not only from rat serum but, for the first time, also brain tissue. Furthermore, we describe animal handling procedures that minimize stress as determined by serum glucocorticoid levels since they can influence cytokine expression. PMID- 15126050 TI - Consistent and reproducible slice selection in rodent brain using a novel stereotaxic device for MRI. AB - Typically small animal radiological images are obtained after placing the animal in the center of the imaging device using beds or platforms, and then adjusting the position after obtaining a scout image. Such a process does not permit the reproducible visualization of the same anatomical plane with repeated examinations. We have developed a device that allows stereotaxic placement of an animal in precisely the same position for repeated examinations. The instrument incorporates a full range of physiological monitoring and life support systems including temperature control, anesthesia delivery and respiratory monitoring. Using magnetic resonance imaging (MRI), the accuracy and reliability of this device is demonstrated in a rat traumatic brain injury (TBI) model. PMID- 15126051 TI - Ca2+-induced Ca2+ release by activation of inositol 1,4,5-trisphosphate receptors in primary pancreatic beta-cells. AB - The effect of sarcoendoplasmic reticulum Ca(2+)-ATPase (SERCA) inhibition on the cytoplasmic Ca(2+) concentration ([Ca(2+)](i)) was studied in primary insulin releasing pancreatic beta-cells isolated from mice, rats and human subjects as well as in clonal rat insulinoma INS-1 cells. In Ca(2+)-deficient medium the individual primary beta-cells reacted to the SERCA inhibitor cyclopiazonic acid (CPA) with a slow rise of [Ca(2+)](i) followed by an explosive transient elevation. The [Ca(2+)](i) transients were preferentially observed at low intracellular concentrations of the Ca(2+) indicator fura-2 and were unaffected by pre-treatment with 100 microM ryanodine. Whereas 20mM caffeine had no effect on basal [Ca(2+)](i) or the slow rise in response to CPA, it completely prevented the CPA-induced [Ca(2+)](i) transients as well as inositol 1,4,5-trisphosphate mediated [Ca(2+)](i) transients in response to carbachol. In striking contrast to the primary beta-cells, caffeine readily mobilized intracellular Ca(2+) in INS-1 cells under identical conditions, and such mobilization was prevented by ryanodine pre-treatment. The results indicate that leakage of Ca(2+) from the endoplasmic reticulum after SERCA inhibition is feedback-accelerated by Ca(2+) induced Ca(2+) release (CICR). In primary pancreatic beta-cells this CICR is due to activation of inositol 1,4,5-trisphosphate receptors. CICR by ryanodine receptor activation may be restricted to clonal beta-cells. PMID- 15126052 TI - Functional separation of deep cytoplasmic calcium from subplasmalemmal space calcium in cultured human uterine smooth muscle cells. AB - For smooth muscle, two important functions of free intracellular calcium (Ca(2+)(i)) are modulation of plasma membrane excitability properties and modulation of the contractile apparatus. As proposed by van Breemen, Ca(2+)(i) can be divided into the subplasmalemmal space (Ca(2+)(sps)) and the deep cytosol (Ca(2+)(d)) by the superficial calcium buffer barrier. Using these distinctions, Ca(2+)(sps) activates the large conductance calcium-activated potassium channel (BK), and Ca(2+)(d) binds calcium-dependent fluorescent probes in the cytoplasm. We present here combined fluorescence-patch clamp experiments designed to simultaneously assess Ca(2+)(d) and Ca(2+)(sps) in cultured human uterine smooth muscle cells. Open probabilities (P(o)) of the BK channel were measured using the cell-attached patch clamp technique. P(o) was used to approximate changes of [Ca(2+)(sps)]. Relative concentrations of Ca(2+)(d) were approximated by observing fluorescence of Calcium green-1 (F). Under control conditions, we found similar time courses for rises of P(o) and F following 10nM oxytocin (OT) addition. In parallel experiments, but with lanthanum (La(3+)) added to the bath to block transmembrane calcium flux, P(o) was only slightly affected, but F increases were delayed and blunted. These data paradoxically indicate that following OT stimulation, the primary source of calcium for Ca(2+)(sps) is internal stores, and calcium entry from the extracellular space is required to raise Ca(2+)(d). When cells were exposed to cyclopiazonic acid (CPA) to release SR calcium stores, P(o) increased slowly, then persisted at large values. The persistence of P(o) rises suggests that removal of calcium from the subplasmalemmal space is primarily via reuptake into the SR. In the presence of La(3+), OT-induced rises of F were slightly prolonged, suggesting that transmembrane calcium flux contributes to decreasing Ca(2+)(d), but is not the primary mechanism. In summary, these data demonstrate that Ca(2+)(d) and Ca(2+)(sps) are not always intimately linked, but indicate a functional separation of the deep cytosol and the subplasmalemmal space that is consistent with the existence of a barrier to calcium diffusion between these two regions. PMID- 15126053 TI - TRPV channels and modulation by hepatocyte growth factor/scatter factor in human hepatoblastoma (HepG2) cells. AB - Using patch clamp and Ca(2+) imaging techniques, we have studied Ca(2+) entry pathways in human hepatoblastoma (HepG2) cells. These cells express the mRNA of TRPV1, TRPV2, TRPV3 and TRPV4 channels, but not those of TRPV5 and TRPV6. Functional assessment showed that capsaicin (10 microM), 4alpha-phorbol-12,13 didecanoate (4alphaPDD, 1 microM), arachidonic acid (10 microM), hypotonic stress, and heat all stimulated increases in [Ca(2+)](i) within minutes. The increase in [Ca(2+)](i) depended on extracellular Ca(2+) and on the transmembrane potential, which indicated that both driving forces affected Ca(2+) entry. Capsaicin also stimulated an increase in [Ca(2+)](i) in nominally Ca(2+)-free solutions, which was compatible with the receptor functioning as a Ca(2+) release channel. Hepatocyte growth factor/scatter factor (HGF/SF) modulated Ca(2+) entry. Ca(2+) influx was greater in HepG2 cells incubated with HGF/SF (20 ng/ml for 20 h) compared with non-stimulated cells, but this occurred only in those cells with a migrating phenotype as determined by presence of a lamellipodium and trailing footplate. The effect of capsaicin on [Ca(2+)](i) was greater in migrating HGF/SF treated cells, and this was inhibited by capsazepine. The difference between control and HGF/SF-treated cells was not found in Ca(2+)-free solutions. 4alphaPDD also had no greater effect on HGF/SF-treated cells. We conclude that TRPV1 and TRPV4 channels provide Ca(2+) entry pathways in HepG2 cells. HGF/SF increases Ca(2+) entry via TRPV1, but not via TRPV4. This rise in [Ca(2+)](i) may constitute an early response of a signalling cascade that gives rise to cell locomotion and the migratory phenotype. PMID- 15126054 TI - The sarcoplasmic reticulum and sarcolemma together form a passive Ca2+ trap in colonic smooth muscle. AB - In smooth muscle, active Ca(2+) uptake into regions of sarcoplasmic reticulum (SR) which are closely apposed to the sarcolemma has been proposed to substantially limit the increase in the cytoplasmic Ca(2+) concentration ([Ca(2+)](c)) following Ca(2+) influx, i.e. the 'superficial buffer barrier hypothesis'. The present study has re-examined this proposal. The results suggest that the SR close to the sarcolemma acts as a passive barrier to Ca(2+) influx limiting [Ca(2+)](c) changes; for this, SR Ca(2+) pump activity is not required. In single voltage-clamped colonic myocytes, sustained opening of the ryanodine receptor (RyR) (and depletion of the SR) using ryanodine increased the amplitude of depolarisation-evoked Ca(2+) transients and accelerated the rate of [Ca(2+)](c) decline following depolarisation. These results could be explained by a reduction in the Ca(2+) buffer power of the cytosol taking place when RyR are opened (i.e. the SR is 'leaky'). Indeed, determination of the Ca(2+) buffer power confirmed it was reduced by approximately 40%. Inhibition of the SR Ca(2+) pump (with thapsigargin) also depleted the SR of Ca(2+) but did not reduce the Ca(2+) buffer power or increase depolarisation-evoked Ca(2+) transients and slowed (rather than accelerated) Ca(2+) removal. However, thapsigargin prevented the ryanodine-induced increase in [Ca(2+)](c) decline following depolarisation. Together, these results suggest that when the SR was rendered 'leaky' (a) more of the Ca(2+) entering the cell reached the bulk cytoplasm and (b) Ca(2+) was removed more quickly at the end of cell activation. Under physiological circumstances in the absence of blocking drugs, it is proposed that the SR limits the [Ca(2+)](c) increase following influx without the need for active Ca(2+) uptake. The SR and sarcolemma may form a passive physical barrier to Ca(2+) influx, a Ca(2+) trap, which limits the [Ca(2+)](c) rise occurring during depolarisation by about 50% and from which the ion only slowly escapes into the main part of the cytoplasm. PMID- 15126055 TI - Time-correlation between membrane depolarization and intracellular calcium in insulin secreting BRIN-BD11 cells: studies using FLIPR. AB - Cytoplasmic Ca(2+) ([Ca(2+)](i)) and membrane potential changes were measured in clonal pancreatic beta cells using a fluorimetric imaging plate reader (FLIPR). KCl (30 mM) produced a fast membrane depolarization immediately followed by increase of [Ca(2+)](i) in BRIN-BD11 cells. l-Alanine (10 mM) but not l-arginine (10 mM) mimicked the KCl profile and also produced a fast membrane depolarization and elevation of [Ca(2+)](i). Conversely, a rise in glucose from 5.6 mM to 11.1 or 16.7 mM induced rapid membrane depolarization, followed by a slower and delayed increase of [Ca(2+)](i). GLP-1 (20 nM) did not affect membrane potential or [Ca(2+)](i). In contrast, acetylcholine (ACh, 100 microM) induced fast membrane depolarization immediately followed by a modest [Ca(2+)](i) increase. When extracellular Ca(2+) was buffered with EGTA, ACh mobilized intracellular calcium stores and the [Ca(2+)](i) increase was reduced by 2-aminoethoxydiphenyl borate but not by dantrolene, indicating the involvement of inositol triphosphate receptors (InsP(3)R). It is concluded that membrane depolarization of beta cells by glucose stimulation is not immediately followed by elevation of [Ca(2+)](i) and other metabolic events are involved in glucose induced stimulus-secretion coupling. It is also suggested that ACh mobilizes intracellular Ca(2+) through store operated InsP(3)R. PMID- 15126056 TI - Calmodulin spatial dynamics in RBL-2H3 mast cells. AB - A line of rat basophilic leukaemia (RBL) cells, a model of mast cells, stably expressing EGFP-tagged calmodulin secreted normally in response to standard agonists. As reported for other cell types, calmodulin was concentrated in the mitotic spindle poles of dividing cells. In unstimulated interphase cells calmodulin was concentrated in the cell cortex and at a single central location. Disruption of cortical actin eliminated the concentration of calmodulin at the cortex while the central calmodulin concentration was associated with an enrichment of tubulin and is likely to represent the centrosome. Following stimulation with either an agonist that crosslinks Fc receptors or co-application of phorbol ester and a calcium ionophore the interior of the cells lost calmodulin while cortical fluorescence became more pronounced but also less uniform. After stimulation discrete bright puncta of calmodulin-EGFP (CaM-EGFP) appeared in the cell interior. Puncta colocalised with moving lysotracker labelled granules, suggesting that calmodulin may play a role in organising their transport. Our results show that in interphase RBL cells a large fraction of the calmodulin pool is associated with targets in the actin cytoskeleton and demonstrate the utility of this model system for studying calmodulin biology. PMID- 15126057 TI - Reduced mitochondrial buffering of voltage-gated calcium influx in aged rat basal forebrain neurons. AB - Alterations of neuronal Ca(2+) homeostatic mechanisms could be responsible for many of the cognitive deficits associated with aging in mammals. Mitochondrial participation in Ca(2+) signaling is now recognized as a prominent feature in neuronal physiology. We combined voltage-clamp electrophysiology with Ca(2+) sensitive ratiometric microfluorimetry and laser scanning confocal microscopy to investigate the participation in Ca(2+) buffering of in situ mitochondria in acutely dissociated basal forebrain neurons from young and aged F344 rats. By pharmacologically blocking mitochondrial Ca(2+) uptake, we determined that mitochondria were not involved in rapid buffering of small Ca(2+) influx through voltage-gated Ca(2+) channels (VGCCs) in the somatic compartment. For larger Ca(2+) influx, aged mitochondria showed a significant buffering deficit. Evidence obtained with the potentiometric indicator, JC-1, suggests a significantly reduced mitochondrial membrane potential in aged neurons. These results support the interpretation that there is a fundamental difference in the way young and aged neurons buffer Ca(2+), and a corresponding difference in the quality of the Ca(2+) signal experienced by young and aged neurons for different intensities of cytoplasmic Ca(2+) influx. PMID- 15126058 TI - Peroxynitrite resistance of sarco/endoplasmic reticulum Ca2+ pump in pig coronary artery endothelium and smooth muscle. AB - We examined the effects of peroxynitrite pre-treatment on sarco/endoplasmic reticulum Ca(2+) (SERCA) pump in pig coronary artery smooth muscle and endothelium. In saponin-permeabilized cells, smooth muscle showed much greater rates of the SERCA Ca(2+) pump-dependent (45)Ca(2+) uptake/mg protein than did the endothelial cells. Peroxynitrite treatment of cells inhibited the SERCA pump more severely in smooth muscle cells than in endothelial cells. To determine implications of this observation, we next examined the effect of the SERCA pump inhibitor cyclopiazonic acid (CPA) on intracellular Ca(2+) concentration of intact cultured cells. CPA produced cytosolic Ca(2+) transients in cultured endothelial and smooth muscle cells. Pre-treatment with peroxynitrite (200 microM) inhibited the Ca(2+) transients in the smooth muscle but not in the endothelial cells. CPA contracts de-endothelialized artery rings and relaxes precontracted arteries with intact endothelium. Peroxynitrite (250 microM) pre treatment inhibited contraction in the de-endothelialized artery rings, but not the endothelium-dependent relaxation. Thus, endothelial cells appear to be more resistant than smooth muscle to the effects of peroxynitrite at the levels of SERCA pump activity, CPA-induced Ca(2+) transients in cultured cells, and the effects of CPA on contractility. The greater resistance of endothelium to peroxynitrite may play a protective role in pathological conditions such as ischemia-reperfusion when excess free radicals are produced. PMID- 15126059 TI - A proton-led model of fast calcium waves. AB - Fast (10-30 microm/s) calcium waves can be propagated through all nucleated eukaryotic cells that have been tested as well as certain cell-free extracts. In a widely used model, they are propagated by a reaction-diffusion cycle in which calcium ions diffuse along the outside of endoplasmic reticula and induce their own release from calsequestrin or calreticulin molecules stored within the reticulum's lumen. Here we propose a new tandem wave model in which they are also propagated by a reaction-diffusion cycle within a reticulum's lumen. In this cycle, increases in luminal [H(+)] induce proton release from luminal calsequestrin or calreticulin. The released protons diffuse ahead to where they release more protons from these luminal storage proteins. What might be called proton induced proton release. They also raise luminal electropositivity. The resultant luminal waves are coordinated with extrareticular ones by movements of calcium and hydrogen ions through the reticular membrane. This model makes five testable predictions which include the autorelease of protons in solutions of calsequestrins or calreticulins as well as waves of increased [H(+)], of increased [Ca(2+)] and of more positive voltage within the reticula of whole cells. Moreover, under some conditions, such luminal waves should cross regions without cytosolic ones. PMID- 15126060 TI - What makes Mona Lisa smile? AB - To study the ability of humans to read subtle changes in facial expression, we applied reverse correlation technique to reveal visual features that mediate understanding of emotion expressed by the face. Surprising findings were that (1) the noise added to a test face image had profound effect on the facial expression and (2) in almost every instance the new expression was meaningful. To quantify the effect, we asked naive observers to rank the face of Mona Lisa superimposed with noise, based on their perception of her emotional state along the sad/happy dimension. Typically, a hundred trials (with 10 or more samples for each rank category) were sufficient to reveal areas altering the facial expression, which is about two orders of magnitude less than in the other reverse correlation studies. Moreover, the perception of smiling in the eyes was solely attributable to a configurational effect projecting from the mouth region. PMID- 15126061 TI - Spatial frequency selective masking of first-order and second-order motion in the absence of off-frequency 'looking'. AB - Converging evidence suggests that, at least initially, first-order (luminance defined) and second-order (e.g. contrast defined) motion are processed independently in human vision. However, adaptation studies suggest that second order motion, like first-order motion, may be encoded by spatial frequency selective mechanisms each operating over a limited range of scales. Nonetheless, the precise properties of these mechanisms are indeterminate since the spatial frequency selectivity of adaptation aftereffects may not necessarily represent the frequency tuning of the underlying units [Vision Research 37 (1997) 2685]. To address this issue we used visual masking to investigate the spatial-frequency tuning of the mechanisms that encode motion. A dual-masking paradigm was employed to derive estimates of the spatial tuning of motion sensors, in the absence of off-frequency 'looking'. Modulation-depth thresholds for identifying the direction of a sinusoidal test pattern were measured over a 4-octave range (0.125 2 c/deg) in both the absence and presence of two counterphasing masks, simultaneously positioned above and below the test frequency. For second-order motion, the resulting masking functions were spatially bandpass in character and remained relatively invariant with changes in test spatial frequency, masking pattern modulation depth and the temporal properties of the noise carrier. As expected, bandpass spatial frequency tuning was also found for first-order motion. This provides compelling evidence that the mechanisms responsible for encoding each variety of motion exhibit spatial frequency selectivity. Thus, although first-order and second-order motion may be encoded independently, they must utilise similar computational principles. PMID- 15126062 TI - Some observations on the effects of slant and texture type on slant-from-texture. AB - We measure the performance of five subjects in a two-alternative-forced-choice slant-discrimination task for differently textured planes. As textures we used uniform lattices, randomly displaced lattices, circles (polka dots), Voronoi tessellations, plaids, 1/f noise, "coherent" noise and a leopard skin-like texture. Our results show: (1) Improving performance with larger slants for all textures, (2) and some cases of "non-symmetrical" performance around a particular orientation. (3) For orientations sufficiently slanted, the different textures do not elicit major differences in performance, (4) while for orientations closer to the vertical plane there are marked differences among them. (5) These differences allow a rank-order of textures to be formed according to their "helpfulness"- that is, how easy the discrimination task is when a particular texture is mapped on the plane. Polka dots tend to allow the best slant discrimination performance, noise patterns the worst. Two additional experiments were conducted to test the generality of the obtained rank-order. First, the tilt of the planes was rotated by 90 degrees. Second, the task was changed to a slant report task via probe adjustment. The results of both control experiments confirmed the texture rank order previously obtained. We then test a number of spatial-frequency-based slant from-texture models and discuss their shortcomings in explaining our rank-order. Finally, we comment on the importance of these results for depth-perception research in general, and in particular the implications our results have for studies of cue combination (sensor fusion) using texture as one of the cues involved. PMID- 15126063 TI - Evaluation of a new quantitative technique to assess the number and extent of preferred retinal loci in macular disease. AB - Patients with scotomas due to macular disease may use more than one preferred retinal locus (PRL) for fixation. We have developed and evaluated an objective, quantitative technique to determine the number of PRLs used during an episode of fixation and the extent of each locus. In five of eight adults with macular disease our techniques consistently indicated the presence of multiple PRLs. Patients with multiple PRLs were more likely to have suffered recent vision loss in the tested eye. Our technique describes fixation more fully than the traditional method of calculating a single bivariate contour ellipse area. PMID- 15126064 TI - Perceptual asynchronies for biological and non-biological visual events. AB - Four experiments investigated the hypothesis that different attributes of a visual scene are processed by independent channels working asynchronously. Experiment 1 considered the attributes of colour, form, and movement of simple geometrical configurations. In each of three conditions, two of these attributes switched simultaneously between two fixed values (Green/Red, Circle/Square, Fixed/Moving). Participants indicated which of the two attributes changes was perceptually closer in time to a sound signal. Response probabilities varied as a function of the time of occurrence of the sound, showing that the processing of the movement channel is delayed with respect to the other two. A smaller but significant difference was also detected between the processing times for colour and form. Comparing Experiments 1 and 2 showed that movement velocity does not affect the delay with which movement onset is perceived with respect to colour. Experiment 3 contrasted colour and movement in the perception of a biological movement. The stimuli were video clips of a coloured ball being lifted by a hand. The colour of the ball changed a variable amount of time before or after the ball started moving. Participants indicated which of the two changes had occurred first. We found that, unlike in Experiments 1 and 2, movement perception no longer lagged colour perception. Experiment 4 tested the hypothesis that the disappearance of the asynchrony is due to perceptual anticipation. We discuss the implications of the results vis-a-vis current theories on perceptual binding and on the coding of dynamic events. PMID- 15126065 TI - Perceptual limits of common fate. AB - We studied the perception of a coherently moving group of collinearly arranged dots ("target dots") that traveled orthogonally to their linear orientation within a background of noise dots moving in random yet straight directions at constant speed ("random-direction noise"). Using a 2-interval forced-choice task we obtained coherence thresholds equal to a signal-to-noise ratio of 1-2%. These thresholds are lower than the 4-10% reported in the literature suggesting that the collinear arrangement of the target dots, in addition to movement, provided form information. Weber's Law was found to hold 4-7 target dots. Overall, sensitivity was constant for a broad range of dot speeds up to at least 6.5 deg/s. Lifetime required for optimal perception was 430 ms, far shorter than the threshold duration of 1 s reported for randomly distributed (i.e., nonaligned) target dots [Vis. Res. 41 (2001) 1891]. Angular deviations from parallel between adjacent motion trajectories were tolerated up to 27 deg for divergence and up to 19 deg for convergence. Diverging motion was detected earlier (after 600-800 ms) than converging motion (>1 s). Forced-choice discrimination yielded a higher proportion of correct responses than the actual (i.e., conscious) perception of the coherently moving group of dots. Our results are consistent with findings from neurophysiological recordings and neuroimaging of motion-sensitive neurons in areas V1 and MT showing broad tuning curves for speed and direction of a moving visual stimulus. PMID- 15126066 TI - The role of salience in localized attentional interference. AB - Observers were cued to attend to two discs from an array and made a discrimination of a target presented within one of the discs. In Experiments 1 and 2, the relative attentional salience of the two attended items was manipulated via the cues (size changes in Experiment 1; size and color changes in Experiment 2). In Experiment 3, the relative salience was manipulated via the luminance contrast of the items themselves. In Experiment 4, relative attentional salience was controlled through a probability manipulation. In all experiments, target performance improved with the relative salience of the target, as well as with increased spatial separation between the two items. This localized interference between cued items varied with visual field. Results are discussed in the context of competition-based models of attentional selection. PMID- 15126067 TI - Improvement of visual acuity by spatial cueing: a comparative study in human and non-human primates. AB - This study investigated the influence of spatial cueing (valid/invalid/no cue) on visual discrimination in human and non-human primates. We employed a spatial resolution task which required the accurate discrimination of the orientation of a Landolt "C" ring. The C appeared as single target in specific retinal locations while subjects maintained fixation of a central fixation point. The minimal discernable size of the "C" (=acuity threshold) was determined as a function of cue condition, retinal eccentricity (3 degrees -15 degrees ), and stimulus onset asynchrony (SOA) (200-1100 ms). For both species, we found consistent benefits from spatial cueing with differences in absolute thresholds ranging from 6% to 25%. These differences increased with retinal eccentricity and decreased with longer SOAs. Further experiments performed with humans only, showed that the effect of spatial cueing on visual discrimination is independent of spatial uncertainty, i.e. the number of possible target locations (2 versus 4), but fades with longer target presentation times. From our results we draw the following conclusions. (i) Since sensory noise and spatial uncertainty was small in our tasks, spatial shifts of attention involve signal enhancement in both, human and non-human primates. (ii) The similarity of the results obtained for humans and macaque monkeys indicates that the latter may serve as a suitable model system in studies trying to tackle the neural underpinnings of attentional control. (iii) In order to elicit robust effects on visual discrimination by spatial shifts of attention, a paradigm comprising short SOAs (approximately 200 ms) and target presentation times (approximately 150 ms), and retinal eccentricities larger than approximately 9 degrees seems most promising. PMID- 15126068 TI - Mechanisms of contrast induction in heterogeneous displays. AB - This study examines how judgments of a region's contrast are influenced by components of a heterogeneous surround. Each stimulus comprised a 5x5 grid of squares in a homogeneous background of fixed mean luminance, with the central square the target. On a given trial, the task was to judge (with feedback) whether the (Weber) contrast of the target was 0.04 or -0.04 (relative to the background); the contrasts assigned (in random order) to the 24 surrounding squares were drawn from the values -0.98, -0.33, 0.33, 0.98 in conformity to one of nine pre-chosen histograms. Presentations were brief (80 ms) in one condition and long (800 ms) in another. A novel psychophysical method was used to estimate the impact exerted on judged target contrast (JTC) by a given contrast in a given grid position. Results were similar for four observers. For both display durations, the four squares sharing an edge with the target influenced JTC 2.4-9 times more than any other surrounding squares. In long presentations, abutting squares of extreme contrast repelled target contrast: squares of contrast -0.98 (0.98) increased (decreased) JTC. However, lower contrast abutting squares attracted target contrast: squares of contrast -0.33 (0.33) decreased (increased) JTC. This central finding can be explained by supposing that: (a) JTC is strongly correlated with the average boundary contrast from surround to target, as registered by linear, edge-selective neurons, and, crucially, (b) the responses of these neurons are themselves subject to lateral inhibition from the rectified responses of other similarly tuned neurons. Finally, in brief presentations, a polarity-specific asymmetry was observed: the two positive abutting-square contrasts continued to influence JTC as they did in long presentations, but contrasts -0.33 and -0.98 ceased to exert much impact, suggesting that lateral influences on target appearance propagate more quickly from positive than from negative contrast abutting regions. PMID- 15126069 TI - Divergent immunological responses following glutaraldehyde exposure. AB - Although Glutaraldehyde (Glut) has been demonstrated to be a moderate contact sensitizer, numerous cases of occupational asthma related to Glut exposure have been reported. The purpose of these studies was to examine the dose-response relationship between Glut exposure and the development of T cell-mediated vs. IgE mediated responses. Initial evaluation of the sensitization potential was conducted using the local lymph node assay (LLNA) at concentrations ranging from 0.75% to 2.5%. A concentration-dependent increase in lymphocyte proliferation was observed with EC3 values of 0.072% and 0.089% in CBA and BALB/c mice, respectively. The mouse ear swelling test (MEST) was used to evaluate the potential for Glut to elicit IgE (1/2 h post challenge) and contact hypersensitivity (24 and 48 h post challenge) responses. An immediate response was observed in animals induced and challenged with 2.5% Glut, whereas animals induced with 0.1% or 0.75% and challenged with 2.5% exhibited a delayed response 48 h post challenge. IgE-inducing potential was evaluated by phenotypic analysis of draining lymph node cells and measurement of total serum IgE levels. Only the 2.5% exposed group demonstrated a significant increase (P < 0.01) in the percentage of IgE(+)B220(+) cells and serum IgE. Following 3 days of dermal exposure, a significant increase in IL-4 mRNA in the draining lymph nodes was observed only in the 2.5% exposed group. These results indicate that the development of an immediate vs. a delayed hypersensitivity response following dermal exposure to Glut is at least in part mediated by the exposure concentration. PMID- 15126070 TI - Molecular analysis of peroxisome proliferation in the hamster. AB - Three novel P450 members of the cytochrome P450 4A family were cloned as partial cDNAs from hamster liver, characterised as novel members of the CYP4A subfamily, and designated CYP4A17, 18, and 19. Hamsters were treated with the peroxisome proliferator-activated receptor alpha (PPARalpha) agonists, methylclofenapate (MCP) or Wy-14,643, and shown to develop hepatomegaly and induction of CYP4A17 RNA, and concomitant induction of lauric acid 12- hydroxylase. This treatment also resulted in hypolipidaemia, which was most pronounced in the VLDL fraction, with up to 50% reduction in VLDL-triglycerides; by contrast, blood cholesterol concentration was unaffected by this treatment. These data show that hamster is highly responsive to induction of CYP4A by peroxisome proliferators. To characterise the molecular basis of peroxisome proliferation, the hamster PPARalpha was cloned and shown to encode a 468-amino-acid protein, which is highly similar to rat and mouse PPARalpha proteins. The level of expression of hamster PPARalpha in liver is intermediate between mouse and guinea pig. These results fail to support the hypothesis that the level of PPARalpha in liver is directly responsible for species differences in peroxisome proliferation. PMID- 15126071 TI - Exposure to 4-tert-octylphenol, an environmentally persistent alkylphenol, enhances interleukin-4 production in T cells via NF-AT activation. AB - 4-tert-Octylphenol (OP) is a representative endocrine disruptor that may have adverse effects on human health. The influence of this compound on allergic immune responses remains unclear. In this study, we have examined the effects of OP on production of interleukin-4 (IL-4), a pro-inflammatory cytokine closely associated with allergic immune responses. OP significantly enhanced IL-4 production in antigen-primed T cells in a dose-dependent manner. Treatment with OP in vivo resulted in significant increase of IL-4 production in T cells and of IgE levels in sera of antigen-primed mice. Furthermore, OP enhanced the activation of IL-4 gene promoter in EL4 T cells transiently transfected with IL-4 promoter/reporter constructs, and the enhancing effect mapped to a region in the IL-4 promoter containing binding sites for nuclear factor of activated T cell (NF AT). Activation of T cells by phorbol-12-myristate-13-acetate (PMA) resulted in markedly enhanced binding activities to the NF-AT site, which significantly increased upon addition of OP, indicating that the transcription factor NF-AT was involved in the enhancing effect of OP on IL-4 production. The enhancement of IL 4 production by OP was blocked by FK506, a calcineurin inhibitor, but not by the estrogen receptor (ER) antagonist ICI 182780. FK506 inhibited the NF-AT-DNA binding activity and IL-4 gene promoter activity enhanced by OP in a dose dependent manner. These findings demonstrate that OP enhances IL-4 production in T cells via the stimulation of calcineurin-dependent NF-AT activation. PMID- 15126072 TI - Vascular effects of ambient particulate matter instillation in spontaneous hypertensive rats. AB - Exposure to ambient particulate matter (PM) is associated with increased mortality and morbidity among those people with cardiovascular impairment. We have studied the effects of exposure to PM or lypopolysaccharide (LPS) on ex vivo vascular function of spontaneous hypertensive rats (SHR) at 4 and 24 h post instillation. Receptor-dependent and -independent relaxation was studied by using acetylcholine (ACh) and sodium nitroprusside (SNP), respectively. We have used phenylephrine (Phe) and KCl for receptor-dependent and -independent contraction. The role of the endothelium was investigated using denuded aorta rings. Exposure to PM (EHC-93, 10 mg/kg) or LPS (350 EU/animal) caused maximal pulmonary inflammation at 24 h post-instillation. PM and LPS elicited a significant increase in receptor-dependent vasorelaxation of aorta compared to saline instilled rats. The largest effect was seen with PM at 4 h post-instillation (EC50 ACh = 2.3 vs. 5 nM control), while at 24 h effects were much smaller (EC50 ACh = 5.6 vs. 5 nM control). SNP-induced vasorelaxation was increased only in EHC 93-treated rats (EC50 = 71.9 vs. 95.7 nM) at 4 h, and this response was higher than that observed at 24 h. Phe induced a dose-dependent vasoconstriction, but no difference was seen between treatments in the presence or absence of endothelium at 4 h. However, at 24 h after instillation of LPS, a right shift of contraction curve was seen (EC50 = 65.3 vs. 43.3 nM). No change was seen in receptor independent vasoconstriction induced by KCl, except in the LPS group at 24 h. A direct relaxation was also observed upon in vitro exposure of aorta rings to PM, and model particles coated with metals. Blood metal analysis showed an increase of zinc and vanadium concentration at 1 and 4 h post-instillation. In conclusion, our data show that PM and LPS instillation has a transient effect on the vasorelaxation of rat aorta that is maximal at 4 h. On the other hand, pulmonary inflammation reaches a maximum at 24 h and coincides with impairment of vasorelaxation. Current data do not allow discriminating among the potential mechanisms, but suggest that both a direct effect of metals and inflammation play a role. PMID- 15126073 TI - The cytotoxicity and mechanisms of 1,2-naphthoquinone thiosemicarbazone and its metal derivatives against MCF-7 human breast cancer cells. AB - We have investigated the antitumor functions and mechanisms of 1,2-naphthoquinone 2-thiosemicarbazone (NQTS) and its metal complexes (Cu(2+), Pd(2+), and Ni(2+)) against MCF-7 human breast cancer cells. The cells were dosed with these complexes at varying concentrations, and cell viability was measured by a sulforhodamine B (SRB) method. To study mechanisms of action, the complexes were incubated with topoisomerase II (topo II) and supercoiled DNA, linear DNA, nicked open DNA, and relaxed DNA were detected by agarose gel electrophoresis. The results revealed that these complexes are effective antitumor chemicals in inhibiting MCF-7 cell growth, with Ni-NQTS being the most effective among the complexes studied. Our data also indicated that Ni-NQTS is more effective than the commercial antitumor drug, etoposide, based on IC(50) values. The mechanistic study of action showed that metal complexes of NQTS, NQ, and NQTS can only stabilize the single-strand nicked DNA, but not double-strand breakage intermediates. In addition, metal derivatives of these ligands, but not the parent NQ and NQTS, exerted an antagonizing effect on topoisomerase II activity. In summary, chemicals with or without metal derivatives might possess different chemical-topoisomerase II-DNA interactions. PMID- 15126074 TI - Environmental contaminant-mixture effects on CNS development, plasticity, and behavior. AB - Environmental contaminants within the polycyclic aromatic hydrocarbon (PAH) and halogenated aromatic hydrocarbon class have been shown to cross the placenta exposing the fetus to the contaminant body burden of the mother. Consequently, a gestational exposure to environmental contaminants may result in increased adverse health outcomes, possibly affecting cognitive performance. Benzo(a)pyrene [B(a)P] and 2,3,7,8, tetrachlorodibenzo-p-dioxin (TCDD) are two prototypical environmental contaminants. A systematic review of the literature suggests that there may be a relationship between vulnerability in susceptible populations and health disparities. The purpose of this mini-review is to provide a point of reference for neurotoxicological studies of environmental contaminant mixture effects on indices of development in general, and on neurodevelopment in particular. Environmental contaminant-mixture-induced decrements in (1) birth index, (2) N-methyl-D-aspartate receptor (NMDA) mRNA expression, (3) long-term potentiation (LTP), (4) fixed-ratio performance learning behavior, and (5) experience-dependent activity related cytoskeletal-associated protein (Arc) mRNA and protein expression collectively support associations between neurobehavioral deficits and gestational exposure to environmental levels of these contaminants. Collectively, data are presented in this mini-review evaluating the effect of gestational exposure to environmental contaminant-mixtures on specific indices of learning and memory, including hippocampal-based synaptic plasticity mechanisms. These indices serve as templates for learning and memory, and as such, from a vulnerability perspective, may serve as targets for dysregulation during development in susceptible populations that have been disproportionately exposed to these contaminants. Included in this review is also a discussion of the relevance of developing biomarkers for use within the framework of cumulative risk-assessment. PMID- 15126075 TI - The effect of drug dose and drug exposure time on the binding, internalization, and cytotoxicity of radiolabeled somatostatin analogs. AB - BACKGROUND: Creation of protease-resistant somatostatin analogs has allowed development of these peptides as clinically useful drugs. Widespread diagnostic use of radiolabeled somatostatin analogs has enhanced interest in the binding and intracellular distribution of these peptides. The degree of drug internalization and length of drug retention may be critical for drug-induced cytotoxicity. We hypothesized that the ability of a radiolabeled peptide to bind to a cell, be internalized, and induce cytotoxicity is proportional to both the radioligand concentration and the exposure time. MATERIALS AND METHODS: To test this hypothesis, somatostatin receptor-expressing cells (IMR-32) were incubated with (111)In-pentetreotide, a sst 2 preferring somatostatin analogue. Radioligand exposure time and/or concentration were varied. RESULTS: Prolonged exposure to a fixed concentration of radioligand resulted in progressive increases in whole cell binding and internalization over time. Cells exposed to a relatively fixed number of microCi-Hr yielded constant whole cell binding and internalization. Increasing the microCi-Hr resulted in a proportionate increase in binding. Cytotoxicity was also proportional to the dose of radiation regardless of whether the exposure was internalized radiation (microCi-Hr from (111)In-pentetreotide) or from external beam radiation (cGy). CONCLUSION: Both drug exposure time and drug concentration contribute to cell binding and cytotoxicity in this model and their relative contributions are inversely related. PMID- 15126076 TI - Role of leukotrienes on hepatic ischemia/reperfusion injury in rats. AB - BACKGROUND: Leukotrienes (LT), composed of cysteinyl LT (cLT; LTC(4), LTD(4), and LTE(4)) and LTB(4), are potent lipid mediators enhancing the vascular permeability and recruitment of neutrophils, which are common features of hepatic ischemia/reperfusion (I/R) injury. The aim of this study was to investigate whether LT can mediate the liver and lung injuries following hepatic I/R. MATERIALS AND METHODS: Sprague-Dawley rats were subjected to 90 min of partial hepatic ischemia followed by 3, 12, and 24 h of reperfusion. In the hepatic and pulmonary tissues, LT content and the mRNA expression of LT-synthesis enzymes, 5 lypoxygenase (5-LO), LTC(4) synthase (LTC(4)-S), and LTA(4) hydrolase (LTA(4)-H) were measured. Tissue injuries were assessed by plasma ALT, histological examination, and wet-to-dry tissue weight ratios. RESULTS: The cLT content in the hepatic tissue after 12 and 24 h reperfusion was increased 4- to 5-fold compared to controls and this was accompanied by the enhancement of hepatic edema and plasma ALT elevation. There were no significant changes in the mRNA expression of LT-synthesis enzymes in both tissues. LTB(4) levels were not increased despite a significant neutrophil infiltration in both tissues. CONCLUSIONS: These data suggest that cLT are generated in the liver during the reperfusion period and may contribute to the development of hepatic edema and exert cytotoxicity. Factors other than LTB(4) may contribute to neutrophil infiltration. PMID- 15126077 TI - Alpha1-acid-glycoprotein protects against trauma-hemorrhagic shock. AB - BACKGROUND: Recent studies have shown that the acute phase protein alpha(1)-acid glycoprotein (AAG) directly modifies endothelial cell responsiveness and is a crucial factor for maintaining endothelial barrier function. We hypothesized that the addition of AAG to the resuscitation fluid will prevent edema formation, increases circulating blood volume, and reduces tissue inflammation following soft tissue trauma and hemorrhagic shock. MATERIALS AND METHODS: Male Sprague Dawley rats (338 +/- 28 g) underwent a 5-cm midline laparotomy (i.e., induction of soft tissue trauma) and were bled to and maintained at a mean arterial pressure of 35 mm Hg for 90 min. The rats were then resuscitated with four times the shed blood volume with Ringer's lactate containing 200 mg/kg AAG or the same amount of albumin. At 6 h after resuscitation, organ wet-to-dry weight ratios and circulating blood volume (Evans blue dilution) were determined. Neutrophil accumulation (myeloperoxidase activity, MPO) and tissue lipid peroxidation (thiobarbituric acid reactive substances) were also measured in the lungs, liver, and intestine. RESULTS: Administration of AAG during the resuscitation significantly increased circulating blood volume and reduced edema formation, neutrophil accumulation, and lipid peroxidation. Interestingly, concomitant plasma IL-6 levels increased while TNF-alpha levels were not significantly affected. CONCLUSIONS: Since addition of AAG to the resuscitation fluid increased circulating blood volume, reduced edema formation, and neutrophil accumulation following trauma and hemorrhagic shock, supplementation of this acute phase protein appears to be a potential adjunct to prevent capillary leakage in patients undergoing major traumatic injury. PMID- 15126078 TI - Direct visualization of nitric oxide release by liver cells after the arrest of metastatic tumor cells in the hepatic microvasculature. AB - BACKGROUND: Our previous studies have shown that the injection of B16F1 melanoma cells into the mesenteric vein can induce the rapid local release of nitric oxide (NO) in the liver, causing apoptosis of the melanoma cells in the liver sinusoids and inhibiting the subsequent formation of hepatic metastases. In this study, we have investigated the distribution and cellular source of NO in this model. MATERIALS AND METHODS: In situ liver perfusion was established in both wild-type (wt) and endothelial nitric oxide synthase knockout (eNOS KO) C57BL/6 mice. A specific fluorescent NO probe, 4,5-diaminofluorescein diacetate (DAF-2 DA) (5 micromol/L), was perfused into the portal venous system to label the liver tissue. Then, a MitoTracker Orange labeled B16F1 melanoma cell suspension (2 x 10(6) cells/ml) was injected through a portal vein catheter by a peristaltic pump. Images of the liver tissue were taken by confocal microscopy from a selected area to determine the cellular source of NO. For quantification, the fluorescence intensity of this area was measured over time by Fluoview software. RESULTS: Diaminotriazolofluorescein (DAF-2T) fluorescence (indicating NO generation) was detected in hepatic parenchymal cells located in the periportal region in both wt C57BL/6 and eNOS KO C57BL/6 mice and was intensified by increased flow rate in the portal venous system. The B16F1 cells arrested in the periportal sinusoids, corresponding to zone 1 of the hepatic acinus. DAF-2T fluorescence was expressed by both sinusoidal lining cells and hepatocytes at the site of tumor cell arrest. The fluorescence intensity of these cells increased approximately 2-fold over a time of 500 s. In contrast, there was no increase in the fluorescence intensity of the sinusoidal lining cells and hepatocytes in mice perfused with buffer or in eNOS KO mice perfused with B16F1 cells. CONCLUSION: This study demonstrates that NO is produced by hepatic parenchymal cells mainly located in the periportal zones and that the arrest of the B16F1 melanoma cells causes an eNOS-dependent local burst of NO by the sinusoidal lining cells and hepatocytes in the periportal areas. PMID- 15126079 TI - Species difference in cholesterol 7alpha-hydroxylase expression of rabbit and rat liver microsomes after bile duct ligation. AB - BACKGROUND: Bile duct ligation (BDL) produces a good animal model for investigation of the metabolic changes in obstructive jaundice. The aim of this study was to investigate the species difference in expression of cholesterol 7alpha-hydroxylase (7alpha-hydroxylase) in rabbits and rats after BDL. MATERIALS AND METHODS: Ten male New Zealand white rabbits weighing 2.5-3 kg and 12 male Wistar strain rats weighing 250-300 g were used. Half the animals underwent BDL, and half were sham operated (Sham). The animals were sacrificed on day 5 after operation. The livers were harvested, and levels of mRNA and 7alpha-hydroxylase activity were determined. Concentrations of serum bilirubin and bile acids were also measured. RESULTS: In BDL rats, the levels of mRNA were increased 30%, and 7alpha-hydroxylase activity was three times that of the Sham group. In BDL rabbits, however, these values were approximately 60 and 50% lower than the Sham group, respectively. Serum bile acid concentrations increased up to 13 times in BDL rabbits and 70 times in BDL rats over that of the Sham groups. Serum cholesterol and serum total bilirubin concentration also increased after BDL in both animals. CONCLUSION: These results suggest that there is a species difference in the expression of 7alpha-hydroxylase after BDL in rabbits and rats and the reason for this difference is most likely pretranslational regulation. PMID- 15126080 TI - Imaging of murine liver tumor using microCT with a hepatocyte-selective contrast agent: accuracy is dependent on adequate contrast enhancement. AB - INTRODUCTION: A major limitation in using both spontaneous and implanted murine liver tumor models in cancer research is the inability to accurately detect and monitor tumor volume. Because microCT without contrast enhancement cannot accurately distinguish tumor from normal liver, we sought to determine the accuracy of contrast enhanced microCT for monitoring liver tumors in mice, performed with intravenous (i.v.) injection of ITG, a hepatocyte-selective contrast agent. METHODS: Twelve female BALB/c mice were injected with 5 x 10(5) CT26 tumor cells in two sites in the liver on day 0, resulting in 24 liver tumors. On days 3, 5, 7, and 10, three mice per day were injected with ITG (0.1 mL ITG/10 g body weight) by tail vein, followed 4 hours later by imaging with microCT (ImTek, Inc.). ITG is transported selectively to hepatocytes by an apoE receptor-mediated process that results in opacification of normal liver parenchyma after i.v. injection. Contrast enhancement on CT scans was graded as good, fair, or poor. After imaging, mice were euthanized to perform gross and histopathologic correlation of liver tumors with CT images. RESULTS: The mean tumor size on microCT and at histopathologic evaluation was 2.2 and 2.3 mm, respectively (P > 0.05). Regression analysis showed no difference between the CT measured tumor and the actual tumor size (P > 0.05). The overall accuracy for detection of tumor on microCT was 88%, with one false-positive and two false negative readings. All three erroneous readings on CT scan occurred in mice in which the contrast enhancement of the liver was poor due to inadequate i.v. injection. Although the overall sensitivity and specificity was 90% and 75%, respectively, this was highly dependent on the degree of contrast enhancement. CONCLUSIONS: MicroCT with ITG contrast is an excellent means to monitor tumor diameter in murine hepatic tumor models. However, adequate contrast enhancement is critical for accurate imaging. PMID- 15126081 TI - Differential effects of opioid peptides on myocardial ischemic tolerance. AB - BACKGROUND: Opioid peptides, which can induce mammalian hibernation, may provide protection against subcellular and molecular changes during hypothermic myocardial ischemia. This study examined the differential effects of the three known myocyte opioid receptors, Mu (micro), Delta (delta), and Kappa (kappa), in augmenting myocardial ischemic tolerance. METHODS: Control hearts (CH) were compared to hearts pretreated with either the micro-agonist, fentanyl, the delta agonist, DADLE, or delta-antagonist, NTB, or the kappa-agonist, U50488H (U50), or kappa-antagonist, nor-BNI. The percent return of isovolemic developed pressure (LVDP), myocardial oxygen consumption (MVO(2)), and coronary flow (CF) following 2 h of global hypothermic cardioplegic ischemia were recorded in isolated Langendorff perfused hearts. RESULTS: At 45 min of reperfusion, hearts pretreated with either DADLE or U50488H demonstrated significantly improved functional recovery versus controls (P < 0.05) and significantly depressed recovery with NTB or nor-BNI pretreatment (P < 0.05). Pretreatment with fentanyl was not significantly different than controls. Furthermore, DADLE, U50488H, or fentanyl resulted in increased MVO(2) versus controls (P < 0.05). There was no difference in CF between all groups. CONCLUSIONS: This study demonstrates that the micro receptor does not appear to confer a beneficial effect. However, selective delta- and kappa-agonists provide significant myocardial protection. Moreover, hearts pretreated with an opioid antagonist showed a marked decrement in both functional and metabolic integrity. These results taken together would imply a positive and negative constitutive role of delta- and kappa-opioids in the regulation of myocardial ischemic tolerance. This utilization of opioid receptor stimulation may have profound clinical applications. PMID- 15126082 TI - Increased apoptosis is specific for acute rejection in rat small bowel transplant. AB - Apoptosis has been associated with several events in solid organ transplantation, including ischemia/reperfusion (IR) injury and acute rejection. To determine whether apoptosis-profiles may distinguish these two conditions, we analyzed apoptosis rates in a rat orthotopic small bowel transplant (SBT) model. SBT was performed in Lewis rats with either freshly harvested or preserved (4 h, in UW at 4 degrees C) syngeneic and allogeneic (Brown-Norway) grafts. Bowel samples were collected 2 h after reperfusion and on small bowel transplant postoperative days (POD) 1, 4, and 7. Apoptosis was detected by measuring levels of histone associated DNA fragments and caspase 3 expression, and by determining apoptotic body counts. All markers measured 2 h after reperfusion increased profoundly in association with preservation. After a significant decrease on POD 1, apoptosis rates rose again between POD 4 and 7 only in allogeneic grafts. This distinct second increase in apoptosis may be an early and specific sign of acute rejection. PMID- 15126083 TI - Expression of VEGF receptors in cocultured neuroblastoma cells. AB - BACKGROUND: VEGF is best known for its angiogenic properties. We have found that VEGF expression is increased in neuroblastoma cells cocultured with hepatocytes. In addition, we have previously shown that neuroblastoma cells cultured with exogenous VEGF have an increase in the expression of VEGF receptors. Therefore, we hypothesized that the expression of VEGF receptors would be up-regulated in neuroblastoma cells grown in the coculture environment. MATERIALS AND METHODS: Two neuroblastoma cell lines (IMR-32 or SK-N-DZ) are used. These cells are cultured alone and in a coculture system with hepatocytes. Message for VEGF and the VEGF receptors KDR, flt-1, flt-4, neuropilin 1 (NRP-1), and neuropilin 2 (NRP 2) are measured with RT-PCR. Flt-4, NRP-1, and NRP-2 protein expression is measured with Western blot. RESULTS: The receptors KDR and flt-1 are not detected in either cell line in either control or coculture conditions. Message for VEGF and flt-4 is significantly increased in the cocultured IMR-32 cells, while that for NRP-1 and NRP-2 is unchanged in these cells. VEGF and its receptors are unchanged in cocultured SK-N-DZ cells. CONCLUSIONS: Neuroblastoma cells express specific VEGF receptors that are differentially regulated in the different cell lines. These findings suggest that the heterogeneity of neuroblastomas may limit the utility of targeting VEGF and its receptors as sole treatments for the tumor, and that successful therapies will be dependent upon the specific biology of the tumor. PMID- 15126084 TI - Novel cardioprotective effects of pravastatin in human ventricular cardiomyocytes subjected to hypoxia and reoxygenation: beneficial effects of statins independent of endothelial cells. AB - Cardioprotective strategies are needed to prevent perioperative myocardial dysfunction in high-risk patients undergoing cardiac surgery. Despite accumulating evidence that statins exert lipid-independent cardioprotective effects, these have been ascribed primarily to improvements in endothelial function and neutrophil-endothelial interaction. The direct effects of statins on cardiomyocytes (independent of endothelial cells) remain unknown. Using a well characterized model of low-volume hypoxia and reoxygenation, we studied the effects of pravastatin on human ventricular cardiomyocytes. Cardiomyocytes were subjected to 90 min of low-volume hypoxia and 30 min of reoxygenation in the presence and absence of pravastatin (1, 10, and 100 microm) (n = 10 per group). In some experiments, the effects of endothelin (ET) receptor blockade (with bosentan) and nitric oxide synthase (NOS) inhibition (with L-NAME) on pravastatin mediated cardioprotection were evaluated. Cell survival, NO, and ET-1 production and protein kinase Akt activation were determined. Pravastatin treatment prevented cardiomyocyte cell death following simulated hypoxia and reoxygenation (P < 0.01). This effect was mediated via an increase in NO release, decrease in myocyte ET-1 production/action, and an increase in protein kinase Akt activation. We demonstrate, for the first time, novel protective effects of pravastatin in human ventricular cardiomyocytes independent of endothelial cells or other cell types. Statin therapy may restore ischemic hearts to full functional integrity during cardioplegic arrest through a direct effect on cardiomyocyte survival. PMID- 15126085 TI - CRF-receptor 1 blockade attenuates acute posttraumatic hyperglycemia in rats. AB - BACKGROUND: Hyperglycemia and insulin resistance after surgical stress are mediated by a complex neuroendocrine response. The present studies were undertaken to determine whether a corticotropin releasing factor (CRF)-receptor 1 (R1) antagonist, CP-154,526 (CP) could alter trauma-induced effects on blood glucose levels, insulin action on skeletal muscle, and dexamethasone-induced suppression of endogenous glucocorticoid secretion. MATERIALS AND METHODS: We used a standardized experimental model of small intestinal resection in the rat. Studies were performed 2 hours after surgery in four groups of rats (n = 24-48) given vehicle or 40 mg of CP i.p. 1 hour before surgical trauma or only anesthesia (controls). Measurements of (I) b-glucose and p-insulin, corticosterone, and ACTH; (II) glucose transport; (III) phosphatidylinositol 3 kinase (PI 3-K) activity in skeletal muscle; and (IV) the dexamethasone suppression test were performed. RESULTS: Surgery resulted in hyperglycemia, reduced insulin-stimulated glucose transport, and a pathological dexamethasone suppression test. B-glucose levels were attenuated in traumatized rats given CP compared to vehicle (P < 0.05). After surgery, p-corticosterone levels were moderately reduced by CP (P < 0.05) and p-ACTH unchanged by the drug. Glucose transport and PI 3-kinase activity as well as the dexamethasone-suppression test were unaffected by administration of CP. CONCLUSIONS: Hyperglycemia in response to small intestinal resection in the rat could be reduced but not inhibited by CRF-R1 blockade. We hypothesize that CRF action within the central nervous system can regulate the hyperglycemic response to surgical stress via mechanisms other than the pituitary-adrenal axis. Our results also indicate that the hypothalamic stress response after surgical stress is dependent on other factors apart from CRF. PMID- 15126086 TI - Arterial injury repair in nonhuman primates-the role of PDGF receptor-beta. AB - BACKGROUND: This study documents the time course of the response to injury of the saphenous artery in baboons and the role of the platelet-derived growth factor beta. Fundamental differences with the well-characterized rat arterial injury model have been found. MATERIALS AND METHODS: Thirty-eight baboons received a unilateral balloon injury to the saphenous artery and were treated with a chimeric blocking antibody to PDGFR-beta or vehicle control for 7, 14, or 28 days. The arteries were evaluated morphologically and for cell proliferation. RESULTS: Both medial and intimal smooth muscle cell proliferation were elevated 7 days after injury and were back close to baseline at 14 days. Unlike the rat, blockade of PDGFR-beta inhibited medial proliferation over 80% at 7 and 14 days, while intimal proliferation was only inhibited at 14 days (>95%). Also, unlike the rat, the baboon arterial media, as well as the intima, increased in size by 14 days after injury. Blockade of PDGFR-beta completely inhibited both intimal and medial growth at 14 days, but there was less of an effect on intimal growth at 28 days. CONCLUSION: Blockade of PDGFR-beta may be a clinical approach to inhibit intimal hyperplasia in humans, but this study raises concerns about the long-term efficacy of this treatment. PMID- 15126087 TI - Cranial repair using BMP-2 gene engineered bone marrow stromal cells. AB - BACKGROUND: Bone grafts, allografts, and biocompatible artificial bone substitutes all have their shortcomings when used for the repair of cranial bone defects. Tissue engineered bone shows promise as an alternative for the repair of these defects. MATERIALS AND METHODS: Rabbit bone marrow mesenchymal stromal cells (MSCs) were separated from iliac crest aspirates and expanded in a monolayer culture 1 month before implantation. These MSCs were then infected with replication-defective adenovirus-human BMP-2 genes 1 week before implantation. Bilateral critical-size cranial defects were created in the animal with removal of osteoinductive periosteum and dura. MSCs were mixed with alginate UP (ultrapure) to form MSC/polymer construct. MSCs used for the control site were infected with adenovirus beta-galactosidase (beta-gal). After 1 week, 6 weeks, and 3 months, five rabbits from each experimental group were sacrificed and the cranial defect site was examined by histology study. RESULTS: Near-complete repair of the large size cranial defects using the tissue engineered MSC/alginate construct was observed. The H&E stain and von Kossa's staining should better regenerate bone at the experiment site. A statistically significant difference in bone formation was noted by 3D CT imaging at 3 months post-BMP-2 treatment of the cranial defects (0.79 +/- 0.06 versus 0.47 +/- 0.05 cm(2), P < 0.001) but not at 6 weeks (0.36 +/- 0.04 versus 0.33 +/- 0.03 cm(2), P = 0.347). CONCLUSIONS: Near complete repair of large cranial defects can be achieved using tissue engineered bone. The use of newly developed polymers as well as the integration of the stem cell concept with gene medicine is necessary to attain this goal. PMID- 15126088 TI - Nitrosoglutathione modulation of platelet activation and nitric oxide synthase expression in promotion of flap survival after ischemia/reperfusion injury. AB - BACKGROUND: Recent studies have shown that platelets play an important role in the pathogenesis of reperfusion injury. Using an inferior epigastric artery skin flap as a flap ischemia/reperfusion (I/R) injury model, we investigated whether the administration of a nitric oxide (NO) donor, nitrosoglutathione (GSNO), could decrease platelet activation and modulate the NO synthase (NOS) activity of platelets and promote flap survival. METHODS: Thirty minutes before flap reperfusion, normal saline (1 mL), nitrosoglutathione (GSNO 0.2, 0.6, 3 mg/kg), or N(G)-nitro-L-arginine-methyl ester (450 mg/kg) was injected intravenously in 10 rats, respectively. The p-selectin (CD62P) expression of platelet activation was detected by a flow cytometry. Immunohistochemical staining was performed to investigate the CD62P deposition on the microvasculature of the flap vessels. NOS isoform expression in the platelets was evaluated by Western blot. Tissue perfusion was monitored by using laser-Doppler flowmetry. Survival areas were assessed at 7 days postoperatively RESULTS: An optimal dose of GSNO (0.6 mg/kg), significantly decreased in CD62P expression on platelets (P < 0.001) and its deposition on the flap vessels, selectively suppressed iNOS induction of platelet, and significantly improved blood perfusion and the flap survival rate (59.8 +/- 4.9% versus 22.1 +/- 6.1%, P < 0.001). In contrast, the NO synthase inhibitor, N(G)-nitro-l-arginine methyl ester, although inhibiting iNOS expression of platelets, compromised platelet activation, tissue perfusion, and flap survival. CONCLUSION: This study suggests that GSNO can appropriately donate NO to suppress platelet activation and platelet iNOS induction, resulting in less platelet activation, better blood perfusion, and flap survival after I/R injury. PMID- 15126089 TI - Improved skin flap survival after local heat preconditioning in pigs. AB - BACKGROUND: Preconditioning induces the expression of heat shock proteins (HSPs), which can help a cell survive an acute episode of stress. Similar to the induction of HSP expression, the cell protection is independent of the type of stress. The aim of this study was to test in a large, randomized animal model, if skin flap survival may be improved by local heat preconditioning and induction of HSP 70. MATERIALS AND METHODS: Twenty-four hours before surgery, a heating blanket was laid on the buttocks of large white pigs. In the preconditioned group (n = 6), the blanket was warmed up to 43 degrees C for 3 x 30 min, whereas it was kept at room temperature in between the heating episodes as well as in the control animals (n = 6). A random pattern skin flap was raised on both sides of the buttocks. Flap survival was measured clinically. Induction of HSP and apoptosis were assessed quantitatively by immunohistochemistry and TUNEL assay, respectively. RESULTS: Preconditioning reduced flap necrosis from 40 +/- 8% of the total flap surface to 7 +/- 14% (P < 0.01). Induction of HSP was significantly higher in the experimental group (79 +/- 12% versus 42 +/- 13%, P < 0.01), whereas apoptosis in healthy flap tissue was reduced from 30 +/- 11 to 11 +/- 6 cells/visual field (P < 0.01). CONCLUSION: In the present study, necrosis and apoptosis rate of skin flaps could be reduced significantly due to local heat preconditioning. Our results suggest that ischemia-related wound healing complications could be diminished with local heat application, a most simple and least invasive method of preconditioning. PMID- 15126090 TI - Editorial: the new sex in the city. PMID- 15126091 TI - Mining the human genome for new health therapies. AB - The completion of the Human Genome Project heralds advances in determining the foundations of disease and in developing new therapeutic treatments. Tests already exist for the detection of some genetic abnormalities that can cause disease, and more are being developed. In the future, pharmacogenetics will be used to tailor treatment to specific patients. PMID- 15126092 TI - The gender gap: new challenges in women's health. AB - Until recently the focus of women's health research and policy has been on reproductive health, but new public health issues have emerged that must be addressed. Among them are a need to examine the basic biology of sex differences in health and disease; the elimination of disparities in health status and health outcomes among diverse populations of women; and a mandate to care for a growing population of aging women. PMID- 15126093 TI - Fast-track women and the quest for children. AB - Childlessness is a tremendously important issue for American women. Women's reproductive lives remain extremely different from men's so that women cannot simply clone the male competitive model in the labor market. A new model is called for to enhance women's chances of crafting lives they want to lead. PMID- 15126094 TI - Men at risk: occupation and male infertility. AB - There is accumulating evidence that workplace exposure to toxic substances contributes to male infertility. Men suffering from infertility problems may do well to look at their occupations, where exposure to certain substances may be a contributory factor, if not a direct cause, of infertility. Most of the studies to date are either case reports or epidemiological studies (population-based, case-control, or cohort studies). Additional, controlled studies need to be done to ascertain the effects of occupational toxins on male infertility. Until then, men and their employers should work together to minimize exposure to these substances. PMID- 15126095 TI - The use of black cohosh to treat symptoms of menopause. AB - The use of complementary and alternative medicine (CAM) is becoming increasingly popular in the United States. In general, the quality of the evidence supporting the efficacy and safety of most CAM therapies is poor. To provide better care and foster an improved doctor-patient relationship, physicians should become informed about CAM, be able to provide educated advice to their patients, and help them integrate any CAM therapies shown to be safe and effective into their health care. SRM will strive to educate and inform, emphasizing the scientific evidence- or lack of it--when evaluating the safety and efficacy of CAM therapies. The appearance of articles on CAM in SRM should not necessarily be seen as an endorsement of the practice or therapy. CAM therapies should be subjected to the same scientific scrutiny as traditional medicines, while recognizing that existing standards for judging treatments can be vulnerable. PMID- 15126096 TI - Antidepressants and sexual dysfunction. AB - Because antidepressant medication is known to cause sexual side effects, physicians who put their patients on antidepressant therapy should counsel them on the possible sexual side effects. Familiarity with other drugs that lessen these side effects might increase patient adherence to treatment. PMID- 15126097 TI - Protect yourself from malpractice suits. AB - There are many areas in the practice of gynecology that can lead to malpractice lawsuits for a practitioner who is not careful, even if he or she practices good medicine. Potential risks include failure to diagnose cancer, not following up with patients, failure to thoroughly check test results, and not keeping proper documentation. The cautious physician will attend to every problem that a patient presents, make sure that patients follow instructions, keep meticulous documentation, and always follow up to make sure his or her patient is progressing well. PMID- 15126098 TI - Shedding our illusions: a better way of medicine. AB - Caregivers are constantly trying to organize their collective efforts in a way that serves the needs of individual patients. And patients themselves seek active involvement and control in their own care. Disorder in medical practice frustrates achievement of all of these goals. Escaping that disorder requires using information tools designed for two purposes: (A) combining patient data with medical knowledge more effectively than the unaided human mind permits, and (B) organizing the multiple processes involved in patient care. PMID- 15126099 TI - Industry update. Serono: innovative research in reproductive health and infertility. AB - The number of couples delaying childbirth is increasing and one out of five American women is having a first child after the age of 35. As a consequence, infertility is becoming more commonplace and the challenges in reproductive health and infertility are growing. The need for innovative research is as compelling as ever. The keys to research in the field over the next several years are furthering our understanding of the biology of reproduction and targeting unmet medical needs. PMID- 15126100 TI - Matrix metalloproteinases in acute inflammation: induction of MMP-3 and MMP-9 in fibroblasts and epithelial cells following exposure to pro-inflammatory mediators in vitro. AB - Recent studies have demonstrated important pro-inflammatory roles for two matrix metalloproteinases (MMPs)-MMP-3 (stromelysin-1) and MMP-9 (gelatinase B)-in acute lung injury [Am. J. Respir. Cell Mol. Biol. 24 (2001) 1]. A role for MMP-3 in skin inflammation has also been demonstrated [Proc. Natl. Acad. Sci. U. S. A. 96 (1999) 6885]. While leukocytes (neutrophils and macrophages) are known to elaborate these tissue-destructive enzymes, parenchymal cells are also capable of synthesizing MMPs. In the present study, we examined the production of MMP-3 and MMP-9 by rodent lung fibroblasts, type II epithelial cells, and vascular endothelial cells. Dermal fibroblasts were also examined. Cells were examined under control conditions and in response to agonists that induce acute inflammatory tissue injury (IgG-containing immune complexes and lipopolysaccharide [LPS]). In the absence of stimulation, MMP-3 and MMP-9 were not detected or were present at low level. However, upon stimulation with either of the two pro-inflammatory agonists, production of both enzymes occurred in fibroblasts and epithelial cells (though not in endothelial cells). The observation that resident cells in the tissue parenchyma can elaborate MMPs in direct response to pro-inflammatory stimuli provides insight into possible mechanisms by which tissue damage occurs in acute inflammation. PMID- 15126101 TI - CD44 negatively regulates apoptosis in murine colonic epithelium via the mitochondrial pathway. AB - Regulation of epithelial cell proliferation and apoptosis are important determinants of colonic crypt homeostasis, and their dysregulations are key features of colon cancer. In this study, we investigated whether CD44, an adhesion protein overexpressed in colon cancer, plays a role in colonocyte proliferation and apoptosis, and the molecular mechanisms involved in these processes. Using a CD44 knockout mouse model devoid of a gross phenotype, we found that CD44 null colonocytes have alterations at the ultrastructural and molecular levels. Mitochondria in CD44 null colonocytes at the top of the crypt have disrupted cristae. The ratio of anti-apoptotic Bcl-xl to pro-apoptotic Bak was shifted toward apoptosis in CD44 null colon due to decreased Bcl-xl expression. Caspase 9 was upregulated and active in CD44 null colon. Its expression shifted from a location restricted to the top of the control crypts to the whole crypt axis in CD44 null colon. Caspase 3 was also activated in CD44 null colon suggesting that CD44 null colonocytes are apoptotic via the intrinsic pathway. Cell cycle regulators, cyclin A, p21, and pRb protein were abrogated in CD44 null mice. Overall, CD44 negatively regulates apoptosis via the mitochondrial pathway in the colonic epithelium through the regulators/effectors of cell cycle and apoptosis. PMID- 15126102 TI - IL-10 inhibits inflammation but does not affect fibrosis in the pulmonary response to bleomycin. AB - Bleomycin yields pulmonary injury characterized by inflammation that proceeds to fibrosis. The production of IL-10 by pulmonary macrophages is increased in the inflammation that accompanies bleomycin lung injury. In the present study, IL-10 deficient and wildtype mice received 0.075 units of bleomycin intratracheally at day 0 and were sacrificed at day 7 or day 14. At day 7, pulmonary inflammation was increased in IL-10-deficient mice as reflected by increased representation of CD3+ and CD4+ lymphocytes and GR-1+ pulmonary granulocytes in the bronchoalveolar lavage (BAL) fluid. Pulmonary interstitial CD80+ and CD86+ mononuclear cells were increased in situ. At day 14, mononuclear cell inflammation was comparable between groups but pulmonary eosinophils were increased in the wildtype. There was no difference in the degree of pulmonary fibrosis, as judged by histology or lung hydroxyproline content. Lung chemokine expression of MIP-1alpha/beta, MIP-2, and eotaxin was increased at days 7 and 14 with a trend towards increased MCP-1 expression at day 14. The findings suggest an immunomodulatory role for IL-10 in the inflammatory response but not in the pulmonary fibrosis yielded by bleomycin. PMID- 15126103 TI - Specifically associated PCR products probed by coincident detection of two-color cross-correlated fluorescence intensities in human gene polymorphisms of methylene tetrahydrofolate reductase at site C677T: a novel measurement approach without follow-up mathematical analysis. AB - Whole blood samples of known methylene tetrahydrofolate reductase (MTHFR) genotypes from 24 individuals were examined at site C677T. Their amplified DNA products were assessed by two-color fluorescence cross-correlation measurements and agarose gel electrophoresis/capillary gel electrophoresis. DNA subpopulations were identified which were not associated with the proper genotype by primer combinations and cycling conditions called multiplexes. We confirmed that DNA analysis by two-color fluorescence cross-correlation measurements allowed the detection of fluorescence signals specifically associated with the proper genotypes in a mixture of amplified nontarget DNA molecules without DNA sizing. The measurement approach does not require complex, follow-up mathematical analysis and is applicable to any single nucleotide polymorphisms. The simple immunogenetic model showed how the approach works to reveal specific DNA target by preventing detection of nontarget DNA. Under those experimental conditions, a new ultrasensitive, and specific method for clinical immunologists is born. PMID- 15126104 TI - Evidence for the involvement of T cell costimulation through the B-7/CD28 pathway in atherosclerotic plaques from apolipoprotein E knockout mice. AB - Antigen-specific T cell activation is thought to influence the initiation and progression of the atherosclerotic plaque. For the T cell activation program to be functional, it is essential not only to facilitate T cell receptor engagement by antigen-presenting cells (APC), but also to deliver requisite costimulatory signals. The most widely comprehended costimulatory pathway involves the ligation of CD28 expressed on T cells by CD80 and CD86 upregulated on APCs. We hypothesized that signals of costimulation should be localized to plaque areas that correspond to the presence of the provoking autoantigen. Atherosclerotic plaques from apoE knockout (KO) mice at different stages of maturation were studied immunohistochemically by monoclonal antibodies to respective markers of primary and secondary lymphocyte activation. Subsequently, flow cytometry studies were conducted in spleen cells from C57BL/6 and apoE KO mice aiming to determine whether the presence of plaques is associated with increased expression of costimulatory signals. We found that regardless of the maturation stage, CD80 and CD86 were evident within the plaques, and colocalized with the presence of markers of dendritic cells and with expression of the extensively investigated autoantigen-oxidized LDL. FACS analysis studies showed that splenocytes from aged atherosclerotic apoE KO mice exhibited increased expression on B cells (which represent APCs) of CD80 and CD86 as compared to their young apoE or naive C57BL/6 litters that have no plaques. These results suggest that primary T cell activation may assume a functional proinflammatory program within the plaque by involvement of CD28/B7 costimulatory pathway. PMID- 15126105 TI - Expression level of Wnt signaling components possibly influences the biological behavior of colorectal cancer in different age groups. AB - Advancing of age apparently influences the behavior of colorectal cancer (CRC). The pattern of activation and expression of Wnt target genes may influence the behavior of the cancer. In the present study, the level of activation of some elements of Wnt signaling was evaluated and correlated with the patient's age and clinicopathological characteristics of the tumor. Beta-catenin and c-Myc mRNA expressions were evaluated by semiquantitative real-time PCR, and subcellular localization of the beta-catenin protein was evaluated by immunohistochemistry. Patients aged 70-84 tended to have locally advanced disease more frequently than younger patients. The same group of patients also more frequently had high nuclear expression of beta-catenin protein and higher expression of c-Myc mRNA. Beta-catenin mRNA had a rather constant expression with advancing of age. High nuclear expression of beta-catenin and high expression of c-Myc were apparently also correlated with locally advanced disease. We concluded that the level of Wnt signaling activation might influence the behavior of the disease in different age groups. PMID- 15126106 TI - The effect of early and late treatment with the tyrphostin AG-556 on the progression of experimental autoimmune myocarditis. AB - Experimental autoimmune myocarditis (EAM) in rats is a T-cell-mediated disorder; the involvement of TNF-alpha in this disorder has been demonstrated. EAM represents a model for human autoimmune myocarditis, a condition for which no optimal treatment is currently available. Tyrphostins AG-126 and AG-556 were previously shown to reduce TNF-alpha production and its end-organ cytotoxicity, thus proving beneficial in animal models of septic shock and experimental autoimmune encephalomyelitis. To study the effects of AG-126 and AG-556 on EAM, we induced the disorder in male Lewis rats through immunization against myosin and subsequently treated the rats with both agents or the control DMSO both before and after the appearance of myocardial inflammation. AG-556 administered daily for 21 days from the day of EAM induction, significantly reduced the severity of myocarditis. Similarly, AG-556 administered for an additional 10 days after myosin immunization (when signs of inflammation are already present) attenuated the progression of myocarditis, though AG-126 did not. TNF-alpha and IFN-gamma production by in vitro sensitized splenocytes from AG-556-treated rats was significantly diminished as compared with control cells from EAM animals. Thus, AG-556 may represent a novel strategy of ameliorating the progression of myocarditis without non-selectively compromising the immune system. PMID- 15126107 TI - Interferon alpha-2b inhibits negative-strand RNA and protein expression from full length HCV1a infectious clone. AB - We have established a T7-based model system for hepatitis C virus (HCV) 1a strain, which involves the use of a replication-defective adenovirus that carries the gene for T7 RNA polymerase and a transcription plasmid containing full-length HCV cDNA clone. To facilitate high-level expression of HCV, sub-confluent Huh7 cells were first infected with adenovirus containing the gene for the T7 RNA polymerase and then transfected with the transcription plasmid. As a negative control, part of NS5B gene of this clone was deleted which abolishes the HCV RNA dependent RNA polymerase and prevents replication of viral RNA. This model produces high levels of structural (core, E1, E2) and nonstructural proteins (NS5), which were detected by Western blot analysis and immunofluorescence assay. Negative-strand HCV RNA was detected only in the wild-type clone in the presence of actinomycin D, and no RNA was detected with the NS5B deleted mutant control. As a practical validation of this model, we showed that IFN alpha-2b selectively inhibits negative-strand RNA synthesis by blocking at the level of protein translation. The inhibitory effect of IFN alpha-2b is not due reduction of transcription by T7 polymerase or due to intracellular degradation of HCV RNA. This in vitro model provides an efficient and reliable means of assaying negative strand RNA, protein processing, and testing the antiviral properties of interferon. PMID- 15126108 TI - Expression of IAP family proteins in esophageal cancer. AB - Members of the inhibitor of apoptosis protein (IAP) family, including survivin, have been reported to be expressed in many tumors. However, their expression in esophageal cancer has not been clarified completely. We investigated the expression of mRNA for IAP family proteins in samples from esophageal cancers and their adjacent normal mucosa tissues by real-time quantitative RT-PCR. The survivin expression in esophageal cancer was significantly higher than that in normal mucosa (P < 0.05). Other IAP family proteins including cIAP1, cIAP2, NAIP and XIAP tended to show stronger expression in cancer tissue than normal mucosa, although the differences were not significant. As to the histological type of tumor, poorly differentiated squamous cell carcinomas exhibited significantly higher level of expression than well-differentiated carcinomas (P < 0.05). The proportion of apoptotic cells of cancer tissue inversely correlated with the intensity of survivin expression (P < 0.05). Immunohistochemical staining demonstrated cytoplasmic as well as nuclear expression of survivin in esophageal cancer, and further, in situ hybridization analysis demonstrated cytoplasmic expression of mRNA for survivin. The results suggest that the expression of IAP family proteins, especially survivin, may be associated with the biological character of esophageal cancer, such as apoptosis. PMID- 15126109 TI - Estrogen receptor codon 594 and HER2 codon 655 polymorphisms and breast cancer risk. AB - The estrogen receptor (ER) and the human epithelial growth factor receptor 2 (HER2) genes have been implicated in the development and prognosis of breast cancer. Several genetic polymorphic sites in these genes have been identified and associated with the risk of breast cancer. We have investigated the association between the estrogen receptor codon 594 (ACA to ACG) and HER2 codon 655 (ATC to GTC) polymorphisms and breast cancer risk. Genomic DNA from breast cancer patients and control subjects was analyzed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). When allelic frequencies of the ER codon 594 and HER2 codon 655 gene were compared, no significant differences were observed between the patient and control groups. (P = 0.063, OR = 1.55, 95% CI = 0.25-9.41 and P = 0.949, OR = 1.01, 95% CI = 0.55-1.88, respectively). In conclusion, our results support the view that both the ER codon 594 and HER2 codon 655 polymorphisms are not associated with increased risk of breast cancer. PMID- 15126110 TI - Three discrete areas within the chromosomal 8p21.3-23 region are associated with the development of breast carcinoma of Indian patients. AB - Deletion in the 22.9 -Mb chromosomal (chr.) 8p21.3-23 region has been shown to be necessary for the development of breast carcinoma (CaBr). In this study, we have attempted to detect the minimal deleted region(s) in the chr.8p21.3-23 region in 62 primary breast lesions having 56 CaBr tumors and six other breast lesions of Indian patients using 15 microsatellite markers. The loss of heterozygosity (LOH) was observed for at least one marker in 96.4% (54/56) of the CaBr samples. Three discrete minimal deleted regions with high frequencies of LOH (39-65%) were identified in the chromosomal 8p23.1-23.2 (D1), 8p23.1 (D2) and 8p 21.3-22 (D3) regions within 2.03, 0.41, 2.47 Mb, respectively. No significant correlation was observed with the high deleted regions and the different clinicopathological parameters. Interestingly, 51.8% (29/56) CaBr samples showed either loss of chr.8p or interstitial deletions in this arm, indicating the importance of chr.8p in the development of CaBr. The pattern of allelic loss in the bilateral lesions had indicated that the lesions were clonal in origin and probably the deletion in the D3 region was the early event among the D1-D3 regions. Thus, our data have indicated that the D1-D3 regions could harbor candidate tumor suppressor gene(s) (TSGs) associated with the development of CaBr. PMID- 15126111 TI - Chronic morphine-induced loss of the facilitative interaction between vasoactive intestinal polypeptide and delta-opioid: involvement of protein kinase C and phospholipase Cbetas. AB - This laboratory recently demonstrated a multiplicative interaction between the pelvic visceral afferent transmitter vasoactive intestinal polypeptide (VIP) and the delta-opioid receptor (DOR)-selective agonist [D-Pen2,5] enkephalin (DPDPE) to regulate cAMP levels in spinal cord [Brain Res. 959 (2003) 103]. Although DOR activation is required for the manifestation of the VIP-DPDPE facilitative interaction, its relevance to opioid antinociception remains unclear. The current study investigates whether or not the VIP-DPDPE facilitation of cAMP formation is subject to tolerance formation, a hallmark characteristic of opioid antinociception. Chronic morphine exposure abolishes the VIP-DPDPE facilitative interaction, consistent with its relevance to DOR antinociception. However, acute in vitro inhibition of protein kinase C (PKC) reinstates the VIP-DPDPE multiplicative interaction characteristic of opioid naive spinal tissue. This suggests that its chronic morphine-induced loss requires a PKC phosphorylation. PKC phosphorylation negatively modulates phospholipase C (PLC)beta, enzymes intimately associated with phosphoinositide turnover and calcium trafficking. These are essential determinants of acute and chronic opioid effects. Accordingly, the effect of chronic morphine on their state of phosphorylation was also investigated. Central nervous system opioid tolerance is associated with the reciprocal phosphorylation (regulation) of two PLCbeta isoforms, PLCbeta1 and PLCbeta3. However, although chelerythrine reinstates the chronic morphine-induced loss of the multiplicative VIP-DPDPE interaction, it does not alter the associated changes in PLCbeta phosphorylation, possibly indicating different time courses of restitution of function and/or involvement of different kinases for different components of tolerance. These results could provide a mechanistic rubric for understanding positive modulation of opioid antinociception by afferent transmission. PMID- 15126112 TI - Beneficial effects of intraventricularly administered BMP-7 following a striatal 6-hydroxydopamine lesion. AB - The present study was undertaken to investigate the effects of bone morphogenetic protein-7 (BMP-7), also named osteogenic protein-1 (OP-1), on the progression of a striatal 6-hydroxydopamine (6-OHDA) lesion. BMP-7, a member of the transforming growth factor-beta (TGF-beta) superfamily of proteins, has been shown to have protective effects in other animal models of neuronal damage. In this study, male Fischer 344 rats received striatal 6-OHDA lesions followed 1 week later by an intraventricular dose of BMP-7. No significant effect of BMP-7 treatment on spontaneous locomotor activity was observed, however BMP-7 significantly increased the density of tyrosine hydroxylase (TH) immunoreactivity (TH-ir) in the substantia nigra (SN) pars compacta, in the lesioned hemisphere [31.7+/-5.2 (optical density (O.D.) arbitrary units) control vs. 50.2+/-4.3 O.D. BMP-7 treated; p<0.05]. Interestingly, BMP-7 significantly increased TH-ir in the SN of the non-lesioned hemisphere (pars reticulata: 14.8+/-1.19 O.D. control vs. 36+/ 2.6 O.D. BMP-7-treated, p<0.05; pars compacta: 29.0+/-4.9 O.D. control vs. 64.4+/ 6.9 O.D. BMP-7-treated, p<0.001). A significant increase in DA concentration in the contralateral, non-lesioned hemisphere was also noted (113.2 ng/g control vs. 198.2 ng/g BMP-7-treated, p<0.01). In contrast to other intraventricularly administered neurotrophic factors, BMP-7 was not associated with an increase in the sensitivity to pain. These results suggest that BMP-7 is able to act as a dopaminotrophic agent without unwanted side effects and as such may be a useful pharmacological tool in the treatment of Parkinson's disease in humans. PMID- 15126113 TI - Increased clusterin expression in old but not young adult S100B transgenic mice: evidence of neuropathological aging in a model of Down Syndrome. AB - S100B is a calcium-binding protein, localized to astroglial cells, which has a variety of neurotrophic functions, including roles in serotonergic neuronal growth, synaptogenesis dendritic branching and apoptosis. In humans, the gene for S100B is found on chromosome 21, within what is considered the obligate region for Down Syndrome (DS) and levels of S100B are increased in brain of both DS and Alzheimer's Disease (AD). We have been characterizing a transgenic mouse overexpressing this protein and have previously found evidence of pathological changes in brains of the mice. In the current study, we have examined the expression of clusterin, a protein expressed in aging neurons, in the mice at two ages. Our findings show increased clusterin expression in the aged S100B mice compared to their CD-1 controls, a finding we have interpreted as further evidence of pathological brain aging. PMID- 15126114 TI - Estrogen receptor subtypes alpha and beta contribute to neuroprotection and increased Bcl-2 expression in primary hippocampal neurons. AB - Estrogen receptor (ER) mediated neuroprotection has been demonstrated in both in vitro and in vivo model systems. However, the relative contribution by either ER subtype, ERalpha or ERbeta, to estrogen-induced neuroprotection remains unresolved. To address this question, we investigated the impact of selective ER agonists for either ERalpha, PPT, or ERbeta, DPN, to prevent neurodegeneration in cultured hippocampal neurons exposed to excitotoxic glutamate. Using three indicators of neuronal viability and survival, we demonstrated that both the ERalpha selective agonist PPT and the ERbeta selective agonist DPN protected hippocampal neurons against glutamate-induced cell death in a dose-dependent manner, with the maximal response occurring at 100 pM. Further analyses showed that both PPT and DPN enhanced Bcl-2 expression in hippocampal neurons, with an efficacy comparable to their neuroprotective capacity. Collectively, the present data indicate that activation of either ERalpha or ERbeta can promote neuroprotection in hippocampal neurons, suggesting that both receptor subtypes could be involved in estrogen neuroprotection. As ERbeta is highly expressed in the brain and has little or no expression in the breast or uterus, discovery and design of ERbeta selective molecules could provide a strategy for activating the beneficial effects of estrogen in the brain without activating untoward effects of estrogen in reproductive organs. PMID- 15126115 TI - Estradiol enhances light-induced expression of transcription factors in the SCN. AB - The suprachiasmatic nucleus of the hypothalamus (SCN) is the master clock that regulates circadian and seasonal rhythms. Among these, the SCN regulates the phasic release of hormones and provides for the timing of the preovulatory luteinizing hormone (LH) surge necessary for ovulation in females. There is little evidence, however, of sex hormone effects on mechanisms underlying SCN function. This study examined the effects of exogenous administration of estradiol on the light-induced expression of transcription factors in the SCN of female rats. Ovariectomized (OVX) female rats were given estradiol or cholesterol implants and perfused 48 h later. Half of the animals were sacrificed 1 h after the regular onset of light within the colony. The rest had the lights go on 2 h prior to the regular time and perfused 1 h later. Collected brains were sliced and sets of SCN sections were processed for immunoreactivity (ir) detecting the Fos, pCREB, egr-1, CREB binding protein (CBP), and calbindin-D (28K) proteins. Following quantification, statistical analyses demonstrated that estradiol enhanced Fos and p-CREB-ir in the SCN of females that experienced a 2-h phase advance. The phase advance also enhanced calbindin and egr-1-ir, but the expression of these proteins was not affected by estradiol. These results demonstrate that estradiol enhances the levels of transcription factors that precede the expression of clock gene proteins in the SCN in response to advances in the onset of environmental light. These data support the hypothesis that steroid hormones play an important role in the fine tuning of the clock in the face of environmental changes in daylight. PMID- 15126116 TI - Sub-chronic administration of zolpidem affects modifications to rat sleep architecture. AB - Gamma-aminobutyric acidA receptor (GABAAR) modulators constitute the majority of clinically relevant sedative-hypnotics. Animal studies have clearly demonstrated sedative efficacy for these compounds in acute studies. However, relatively less is known regarding their efficacy under brief periods of repeat administration or following intermittent dosing. Therefore zolpidem, a short-acting GABAAR modulator with selectivity for the type-I (omega1) benzodiazepine receptor, was studied for efficacy in altering rat sleep architecture as determined by electrocorticogram (ECoG) and electromyogram (EMG) activity over a 7-day sub chronic administration period. Zolpidem caused significant reductions in wakefulness entries and rapid eye movement (REM) sleep entries and duration, with increases in Delta sleep duration throughout the administration period. Examination of sleep architecture 24 h after cessation of sub-chronic zolpidem administration revealed a decrease in Delta sleep, suggesting that repeated zolpidem administration might elicit enduring modifications to sleep organization. This was not seen following similar dosing of diazepam. The efficacy of sub-chronic administration of zolpidem to alter sleep architecture was enhanced when the administration regimen was repeated following a 7-day hiatus. Significant increases in Delta sleep duration, with significant decreases in light sleep and wakefulness were observed during the repeated exposure to zolpidem. Therefore, sub-chronic administration of zolpidem affected lasting modifications in sleep organization that appeared both 1 day following administration and during reiterated administration without eliciting tolerance. PMID- 15126117 TI - Mediation of BMP7 neuroprotection by MAPK and PKC IN rat primary cortical cultures. AB - We have previously demonstrated that pretreatment with bone morphogenetic protein 7 (BMP7), a trophic factor in the TGFbeta superfamily, reduces ischemia-induced brain infarction induced by middle cerebral artery ligation in rats. Since the mitogen-activated protein kinase (MAPK) pathway is involved in many TGFbeta mediated responses, we examined the interaction of BMP7 and MAPK in primary cultures obtained from the cerebral cortex of E16-17 rat embryos. Lactate dehydrogenase (LDH) in the media was used as an index of cell death. BMP7 did not alter LDH levels at low concentration (1.25 nM), but exhibited increased cellular toxicity at higher concentration (>12.5 nM). BMP7 at the low concentration significantly attenuated H2O2-induced increases in LDH activity and decreases in neuronal density. Pharmacological interactions were used to examine if MAPK was involved in this response. BMP7-induced protection was antagonized by the p42,44 MAPK kinase inhibitors PD98059 and U0125. The p38 MAPK antagonist SB203580, and their inactive analog SB202474, also attenuated BMP7-induced protection, suggesting that the interaction with p38 MAPK is nonspecific. Previous studies have indicated that SB202474 has inhibitory effects on other protein kinases. We found that the protein kinase C inhibitor chelerythrine antagonized BMP7-induced protection against H2O2. Western blot analysis indicated that BMP7 increased phosphorylation of p42,44 MAPK and PKC. Taken together, our data suggest that BMP7 is neuroprotective at low concentrations in primary cortical cell culture. The protective effects of BMP7 may involve the activation of p42,44 MAPK and PKC. PMID- 15126118 TI - Role of prostaglandin receptor subtype EP1 in prostaglandin E2-induced nociceptive transmission in the rat spinal dorsal horn. AB - It has been indicated that prostaglandin E2 (PGE2) and the receptor for PGE2 (EP receptor) are key factors contributing to the facilitated generation of nociception. This study was designed to investigate the roles of PGE2 and EP1 receptors in the spinal cord in the nociceptive transmission, using behavioral and intracellular calcium ion concentration ([Ca2+]i) assays and in situ hybridization. Experiments were conducted on Sprague-Dawley rats. In behavioral assays, withdrawal thresholds to mechanical stimuli were evaluated using von Frey filament. The effect of an intrathecally administered selective EP1 antagonist, 6 [(2S,3S)-3-(4-chloro-2-methylphenylsulfonylaminomethyl)-bicyclo[2.2.2]octan-2-yl] 5Z-hexenoic acid (ONO-8711), on the intrathecal PGE2-induced hyperalgesia was examined. In [Ca2+]i assays, we measured [Ca2+]i in the dorsal horn of spinal cord slices and examined the effects of PGE2 and ONO-8711 perfusion on the [Ca2+]i changes. In situ hybridization using EP1 digoxigenin probe was performed on the slice sections of the lumbar spinal cord and bilateral L4 and L5 dorsal root ganglions (DRGs). Mechanical hyperalgesia was observed after intrathecal PGE2 administration. Intrathecal administration of ONO-8711 attenuated the PGE2 induced mechanical hyperalgesia in a dose- and time-dependent manner. Perfusion of ONO-8711 markedly suppressed PGE2-induced [Ca2+]i increment in laminae II-VI in dorsal horn of the spinal cord slice. Moreover, in situ hybridization revealed EP1 hybridization signals in the DRG neurons, but not in the spinal cord. The results of this study suggested that spinal PGE2 activates the EP1 receptors existing on the central terminals of primary afferents, subsequently increasing in [Ca2+]i in the spinal dorsal horn, which are involved in the mechanisms of spinal PGE2-induced nociceptive transmission. PMID- 15126119 TI - Cell type- and brain structure-specific patterns of distribution of minibrain kinase in human brain. AB - The minibrain kinase (Mnb/Dyrk1A) gene is localized in the Down syndrome (DS) critical region of chromosome 21. This gene encodes a proline-directed serine/threonine protein kinase (minibrain kinase-Mnb/Dyrk1A), which is required for the proliferation of distinct neuronal cell types during postembryonic neurogenesis. To study the distribution of Mnb/Dyrk1A during human brain development and aging, we raised Mnb/Dyrk1A-specific antibody (mAb 7F3) and examined 22 brains of normal subjects from 8 months to 90 years of age. We found that neurons were the only cells showing the presence of 7F3-positive product in both cell nucleus and cytoplasm. Nuclear localization supports the concept that Mnb/Dyrk1A may be involved in control of gene expression. Synaptic localization of Mnb/Dyrk1A also supports our previous studies suggesting that Mnb/Dyrk1A is a regulator of assembly of endocytic apparatus and appears to be involved in synaptic vesicle recycling and synaptic signal transmission. Accumulation of numerous 7F3-positive corpora amylacea in the memory and motor system subdivisions in subjects older than 33 years of age indicates that Mnb/Dyrk1A is colocalized with markers of astrocyte and neuron degeneration. Differences in the topography and the amount of Mnb/Dyrk1A in neurons, astrocytes, and ependymal and endothelial cells appear to reflect cell type- and brain structure-specific patterns in trafficking and utilization of Mnb/Dyrk1A. PMID- 15126120 TI - Antipsychotic drugs clozapine and olanzapine upregulate bcl-2 mRNA and protein in rat frontal cortex and hippocampus. AB - To investigate whether atypical antipsychotic drugs exert an effect on the neuroprotective protein Bcl-2, rats were chronically treated with clozapine or olanzapine for 28 days. The expression of bcl-2 mRNA and Bcl-2 protein was determined, respectively, by in situ hybridization in tissue sections and Western blot on tissue samples from rat frontal cortex and hippocampus. The bcl-2 mRNA positive cells were increased in these regions after drug treatments, and quantitative analysis revealed that the upregulation of bcl-2 mRNA and Bcl-2 protein is significantly greater than in vehicle control (p<0.05 or p<0.01). It is demonstrated that neuroprotective protein Bcl-2 is involved in the action of atypical antipsychotic drugs, and might represent a neuroprotective mechanism of these drugs. PMID- 15126121 TI - Disparity sensitivity in the superior colliculus of the cat. AB - The present study aims at evaluating the spatial disparity response profiles of binocular cells in the superficial layers of the superior colliculus of the cat using drifting light bars and phase-shifted spatial frequency gratings. Results show that a total of 64% of the cells were sensitive to phase disparities and had large tuning profiles. Similarly, a large proportion (75%) of those tested with position offsets showed one of the four classic disparity profiles, those of the tuned cells being rather coarse. When tested with both position and phase disparities, 54% of the cells showed sensitivity profiles to the two types of stimuli. The overall results suggest that the superior colliculus is involved in the analysis of coarse stereopsis and/or the planning and initiation of saccades during vergence eye movements and/or the control of fine adjustments to maintain fixation as the stimulus moves in depth. PMID- 15126122 TI - Morphology and electrophysiology of neurons in dog paraventricular nucleus: in vitro study. AB - The paraventricular nucleus (PVN) of the hypothalamus plays an important role in regulating gut motility. To date, there have been no intracellular electrophysiological studies of dog PVN neurons in vitro. The aims of this study were to: (1) adapt brain slice methods developed for studies of rodent CNS tissue to canine CNS tissue; and (2) study the electrophysiology and morphology of single neurons of the dog paraventricular nucleus (PVN). Coronal hypothalamic slice preparations (400 microm thick) of dog brain were used. Three groups of PVN neurons were classified based on their firing pattern. Continuous firing neurons (n=32) exhibited continuous ongoing action potentials (APs). Burst firing neurons generated bursts of APs (n=19). Intermittent firing neurons had only a few spontaneous APs. In contrast to continuous firing neurons, 14 of 19 burst firing neurons and 3 of 7 intermittent firing neurons responded to depolarizing current with a Ca2+-dependent low-threshold potential. Twenty-one PVN neurons studied electrophysiologically were filled with biocytin. Continuous firing neurons (n=12) had oval-shaped soma with two or three sparsely branched dendrites. Branched axons were found in two continuous firing neurons, in which one branch appeared to terminate locally. Burst firing neurons (n=8) generally had triangular soma with 2 to 5 branched dendrites. In summary, the brain slice technique was used to study the morphology and electrophysiology of single neurons of the dog brain. Electrophysiological and morphological properties of the three neuron groups were identified and discussed. PMID- 15126123 TI - Bone marrow grafts restore cerebral blood flow and blood brain barrier in stroke rats. AB - We monitored alterations in cerebral blood flow (CBF) and blood-brain barrier (BBB) permeability following middle cerebral artery occlusion (MCAo) and intrastriatal transplantation of mouse bone marrow stromal cells (BMSCs) or saline infusion in adult Sprague-Dawley rats. Laser Doppler and Evans Blue assay revealed that BMSC grafts dose-dependently restored CBF and BBB to near normal levels at a much earlier period (Days 4-5 post-MCAo) in transplanted stroke animals compared to stroke animals that received saline infusion (Days 11-14 post MCAo). Xenografted BMSCs survived in the absence of immunosuppression, and elevated levels of transforming growth factor-beta superfamily of neurotrophic factors were detected in transplanted stroke animals. These data suggest that early restoration of CBF and BBB following transplantation of BMSCs could mediate the reported functional outcomes in stroke animals. PMID- 15126124 TI - Sustained potentiation by substance P of NMDA-activated current in rat primary sensory neurons. AB - This study aimed to explore the modulatory effect of substance P (SP) on the current response mediated by N-methyl-D-aspartate (NMDA) receptor in rat primary sensory neurons and its time course using whole-cell patch clamp technique. The majority of neurons (179/213, 84.0%) examined were sensitive to NMDA (0.1-1000 microM) with an inward current, and a proportion of the NMDA-sensitive neurons also responded to SP (78/98, 80.0%) with an inward current. Pretreatment with SP potentiated the NMDA-activated current (INMDA) in a non-competitive manner, which is shown in that SP shifted the concentration-response curve for NMDA upwards compared with the control; the maximal value of INMDA increased fourfold, while the EC50 values for both curves were very close (28 vs. 30 microM). Furthermore, this potentiating effect was time-dependent: the amplitude of INMDA reached its maximum 20 min after SP preapplication, and thereafter maintained a steady level of about 2-3 times its control for 2 or even 3 h. This sustained potentiation by SP of INMDA could be blocked by extracellular application of WIN51708, a selective non-peptide antagonist of NK-1 receptor; and abolished by intracellular application of either BAPTA, or H-7, or KN-93. Though NMDA applied alone also induced a short-term (less than 20 min) self-potentiation of INMDA, it could be abolished by intracellular dialysis of BAPTA or KN-93 completely. As is known, the cell body of dorsal root ganglion (DRG) neurons is generally used as an accessible model for studying the characteristics of the membrane of primary afferent terminals in the dorsal horn of spinal cord. Therefore, these results may offer a clue to the explanation of the symptoms of chronic pain. PMID- 15126125 TI - Modulation of sodium currents in rat dorsal root ganglion neurons by sulfur dioxide derivatives. AB - The effect of sulfur dioxide (SO2) derivatives, a common air pollutant and exists in vivo as an equilibrium between bisulfate and sulfite, on tetrodotoxin sensitive (TTX-S) and tetrodotoxin-resistant (TTX-R) sodium channels in cultured post-natal dorsal root ganglion (DRG) neurons were studied using the whole cell configuration of patch-clamp technique. SO2 derivatives on two types of sodium currents were either inhibitory or stimulatory depending on the kinetic parameters tested. At a holding potential of -80 mV, SO2 derivatives suppressed TTX-S sodium currents when depolarizing potential was negative to -30 mV and TTX R sodium currents when negative to -10 mV but they increased them when the depolarizing potential was positive to -30 or -10 mV. SO2 derivatives shifted the conductance-voltage curve for TTX-R sodium currents in the depolarizing direction but had little effect on that for TTX-S sodium currents. The steady-state inactivation curve for TTX-R sodium channel was shifted by SO2 derivatives in the depolarizing direction as that for TTX-S sodium channel. SO2 derivatives changed the reversal potential and increased the maximum conductance of two types of sodium channels. SO2 derivatives postponed the activating time and delayed the inactivation of sodium currents. The results suggest that SO2 derivatives would increase the excitability of neurons and alter the ion selectivity for two types of sodium currents. PMID- 15126126 TI - Strain-related variations of AMPA receptor modulation by calcium-dependent mechanisms in the hippocampus: contribution of lipoxygenase metabolites of arachidonic acid. AB - Several studies have demonstrated that C57 and DBA mice exhibit behavioural differences in diverse learning tasks as well as variations in the expression of long-term potentiation (LTP) in the hippocampus. In the present investigation, we tested the possibility that these differences between the two strains might be attributable to differential regulation of hippocampal alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate (AMPA) receptors by calcium-dependent mechanisms. Using in vitro receptor autoradiography, we found that calcium treatment of C57 mice sections resulted in a marked increase of 3H-AMPA binding in areas CA3 and CA1 of the hippocampus and in the dentate gyrus. However, we discovered that the ability of calcium to upregulate 3H-AMPA binding in the DBA strain was much lower than in corresponding regions from the C57 strain. Western blot and immunohistochemical experiments indicated that truncation of AMPA receptor subunits by calcium-dependent mechanisms was possibly not responsible for the binding differences, as no significant variations in glutamate receptor subunit 1 (GluR1) and GluR2/3 immunoreactivity were observed between the two strains after calcium treatment. Interestingly, we found that strain-related variations in the regulation of 3H-AMPA binding by calcium were totally eliminated when brain sections were preincubated with preferential inhibitors of lipoxygenase (LO) pathways of arachidonic acid (AA) metabolism. Taken together, these results suggest that calcium-induced regulation of AMPA receptors varies between the two strains and that this variation might be linked to the production of specific AA metabolites. PMID- 15126127 TI - Effect of etodolac, a COX-2 inhibitor, on neuropathic pain in a rat model. AB - Etodolac, a cyclooxygenase-2 inhibitor, may alleviate nociceptive pain and inhibit the activation of osteoclasts. The aim of the present study was to determine whether etodolac can alleviate heat-evoked hyperalgesia and investigate its possible protective effects on osteoporosis induced by chronic constriction injury (CCI) in rats. A CCI to the sciatic nerve was performed, after which the rats received etodolac orally in a volume of 2 ml at 0, 1, and 10 mg/kg/day for 1 to 5 weeks following surgery (experiment 1); at 0 and 10 mg/kg/day for 1 day to 5 weeks following surgery (experiment 2); and at 0 mg/kg/day for 1 to 5 weeks, 10 mg/kg/day for 1 to 2 weeks after surgery, or 10 mg/kg/day for 1 to 3 weeks after surgery (experiment 3). Paw withdrawal latency after exposure to heat, bone mineral content (BMC) and bone mineral density (BMD) in the whole tibial bone, and the number of tartrate resistant acid phosphate (TRAP)-positive multinucleated osteoclasts were measured. Etodolac alleviated heat-evoked hyperalgesia in the CCI rats and the increase in number of TRAP-positive multinucleated osteoclasts on the CCI-side was abrogated, however, it did not inhibit the decrease of BMC and BMD on the CCI-side. Our results suggest that etodolac is useful for treatment of neuropathic pain. PMID- 15126128 TI - Stimulation of the superior cerebellar peduncle during the development of amygdaloid kindling in rats. AB - Cerebellar manipulations have been used successfully in some intractable epileptic patients, however, their intrinsic mechanisms are not fully understood. To further clarify the cerebellar participation in epilepsy, we stimulated 10 rats with 100 Hz, 20 microA at the superior cerebellar peduncle (SCP) during amygdaloid kindling. Results were compared to 10 rats with an electrode placed at the SCP without stimulation and 10 rats without electrodes at the SCP used as control. We found that SCP stimulation increased the theta and alpha rhythms at the contralateral motor cortex. Such a stimulation produced hypertonicity of the forelimbs and tremor of the head. In this condition, we found that each of the behavioral stages during amygdaloid kindling in the SCP stimulated rats was reached earlier, while the amygdaloid electrographic afterdischarges (ADs) were longer during the first and shorter in the final trials as compared to controls. Moreover, amygdaloid ADs recorded exclusively during the behavioral stage-5 were significantly shorter than those recorded in the control conditions. We suggest that SCP stimulation could change the customary electrographic and convulsive expression of amygdala kindling in such a manner as to initially facilitate the limbic seizures and impede the secondary generalized seizures. PMID- 15126129 TI - Chemical identity of 5-HT2A receptor immunoreactive neurons of the rat septal complex and dorsal hippocampus. AB - Brain 5-HT2A receptors have been implicated in various behavioural and physiological processes including hippocampus-dependent learning and memory. To clarify the cellular localization and chemical identity of 5-HT2A receptor immunoreactive (-ir) neurons in the rat septal complex and dorsal hippocampus, an immunofluorescence histochemical study was performed using a monoclonal antibody to the 5-HT2A receptor. Pretreatment with colchicine increased the number of 5 HT2A receptor-ir cell bodies, indicating that the 5-HT2A receptor protein undergoes microtubule-dependent anterograde transport in axons and dendrites. 5 HT2A receptor immunoreactivity was detected in septal cholinergic neurons, identified with an antiserum to the vesicular acetylcholine transporter (VAChT), and in GABAergic cell bodies in the medial septum/diagonal band of Broca, identified with antisera to glutamic acid decarboxylase (GAD) and the calcium binding protein parvalbumin. In the dorsal hippocampus, 5-HT2A receptor immunoreactivity was demonstrated in cells located in the pyramidal cell layer (CA1-3) throughout the Ammon's horn and in the granular cell layer of the dentate gyrus. Furthermore, 5-HT2A receptor immunoreactivity was present in most hippocampal interneurons identified by the presence of GAD65, parvalbumin, calbindin D-28k, somatostatin and neuropeptide Y. In contrast, 5-HT2A receptor immunoreactivity was present in only a few interneurons containing cholecystokinin and calretinin immunoreactivity. The results suggest that serotonin acting on 5-HT2A receptors can modulate hippocampal functions via direct actions on hippocampal glutamatergic principal cells and indirectly via actions on hippocampal interneurons with different phenotypes as well as GABAergic and cholinergic septohippocampal neurons. PMID- 15126130 TI - The distribution of serotonin transporter immunoreactivity in hippocampal formation in monkeys and rats. AB - To elucidate the anatomical distribution of the serotonergic neurotransmitter system, we identified serotonin transporter (5-HTT) in the hippocampus of rats and monkeys by immunohistochemistry. A widespread and heterogeneous distribution of 5-HTT-immunoreactive fine fibers was noted in the rat brain. However, in monkeys, punctuate 5-HTT-immunoreactive deposits and fewer fibers were observed. The species difference in 5-HTT immunohistochemical staining pattern may be caused by differences in localization of 5-HTT between species. PMID- 15126131 TI - Serotonin innervation of the primate suprachiasmatic nucleus. AB - The suprachiasmatic nucleus (SCN) in rodents receives a dense innervation from serotonin neurons of the midbrain raphe. This projection overlaps the terminal field of the retinohypothalamic tract in the SCN core, the central part of the nucleus characterized by a population of vasoactive intestinal polypeptide (VIP) containing neurons. To determine whether a similar pathway is present in primates, we carried out an immnunocytochemical investigation of the primate SCN using antisera against either serotonin (monkey) or the serotonin transporter (human). This demonstrated a dense serotonergic plexus over the SCN core in both species. As in rodents, the distribution of the serotonin innervation of the primate SCN overlaps that of the retinohypothalamic input and the VIP neuronal population. We also find a supraependymal plexus of serotonin axons in the third and lateral ventricles of the human and monkey brains that is similar in distribution, but less dense, than the one reported in rodents. PMID- 15126132 TI - Effects of furosemide and bendroflumethiazide on saliva flow rate and composition. AB - Aim of this study was to evaluate the effect on saliva flow rate and composition and on perceived xerostomia. The study used a Latin square design, all subjects being once daily (at 7.00 a.m.) taking the bendroflumethiazide (2.5 mg), furosemide (40 mg), or placebo, in a randomised order. Each treatment period of 7 days was separated by wash-out periods of 14 days. Unstimulated and paraffin chewing stimulated whole saliva, and 3% citric acid stimulated parotid and submandibular-sublingual secretion were collected twice daily, at 7.30 a.m., with the patients in a fasting condition (morning values), and at 10.30 a.m., about 2 h after intake of a standard breakfast (lunchtime values), on day 0 (baseline), day 1 (acute treatment), and day 7 (chronic treatment). Saliva flow rates were measured and all four secretions were analysed for the concentration of sodium, potassium, chloride, and total protein. Xerostomia was assessed by means of a Visual Analogue Scale. Statistical analysis used the Wilcoxon signed rank test. For flow rate, only that of submandibular-sublingual secretion was affected, significantly so in the morning during chronic treatment with both drugs. In resting whole saliva the output of both sodium and chloride tended to decrease especially during treatment with bendroflumethiazide, while in submandibular sublingual secretion the output of all the electrolytes was decreased, especially for potassium and chloride and during treatment with furosemide. Further, xerostomia tended to increase during treatment with furosemide, statistically significant at lunchtime during chronic treatment. In conclusion, this study has demonstrated a modest effect on salivary flow rate and a more pronounced effect on saliva composition, especially in submandibular-sublingual secretion during treatment of healthy volunteers with therapeutic doses of two different diuretics, encouraging clinical studies in hypertensive patients and basic research as to the presence of a thiazide sensitive Na-Cl cotransporter in human salivary glands. PMID- 15126133 TI - Biochemical analysis of saliva and taste acuity evaluation in patients with burning mouth syndrome, xerostomia and/or gustatory disturbances. AB - OBJECTIVE: In the current study, we performed taste and salivary analysis on patients suffering from burning mouth syndrome and xerostomia or taste disturbances. STUDY DESIGN: A total of 180 patients who complained of idiopathic burning mouth syndrome (BMS) and taste aberrations and/or xerostomia that may accompany BMS were evaluated. These patients were compared with 90 healthy, age- and sex-matched controls. Salivary flow rate, biochemical and immunological analysis and taste acuity by the forced-choice drop technique were performed for all subjects. These analyses were found to be conclusive in distinguishing controls from patients with complaints. RESULTS: The great similarity of both salivary and taste analysis in the BMS, taste aberration and xerostomia groups, which were significantly different from the results obtained in the control group, was found to be the most striking result. Higher salivary concentrations in the experimental group were consistent with a lower saliva (water) flow rate. CONCLUSION: An oral neuropathy and/or neurological transduction interruption induced by salivary compositional alterations is suggested as the possible aetiology for the complaints. This report may add an important objective diagnostic tool to the clinician treating these patients. PMID- 15126134 TI - Human salivary function in relation to the prevalence of Tannerella forsythensis and other periodontal pathogens in early supragingival biofilm. AB - OBJECTIVE: Previously, we screened 149 subjects and established four groups high or low for salivary killing of oral bacteria, and for aggregation and live and dead adherence of oral bacteria (as a combined factor). Caries scores were significantly lower in both High Aggregation-Adherence groups. Subsequently, we found that supragingival total biofilm DNA, total streptococci and two major streptococcal rRNA variants also were significantly lower in the High Aggregation Adherence groups. In this study, we looked at the effects of those differences in salivary function on three periodontal pathogens. DESIGN: Quantitative PCR was used to determine levels of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis and Tannerella forsythensis (formerly Bacteroides forsythus) in stored DNA extracts of overnight supragingival biofilm collected from buccal upper central incisors (UC), lingual lower central incisors (LC) and buccal upper and lower first molars (BM) and lingual upper and lower first molars (LM) of subjects in the four groups. RESULTS: A. actinomycetemcomitans and P. gingivalis were almost completely absent from these samples. T. forsythensis was found in 11 of 35 persons at the buccal molar site. Only two of those subjects were in the High Aggregation-Adherence groups, and that difference was statistically significant. The mean quantity of T. forsythensis also was significantly lower in the High Aggregation-Adherence groups. CONCLUSIONS: The difference between the Low and High Aggregation-Adherence groups might reflect direct interactions of salivary proteins with T. forsythensis. Alternatively, the higher levels of total biofilm and total streptococci seen in the Low Aggregation-Adherence groups might create a favourable environment for early secondary colonization of T. forsythensis. PMID- 15126135 TI - Morphological basis of sensory neuropathy and neuroimmunomodulation in minor salivary glands of patients with Sjogren's syndrome. AB - OBJECTIVE: A predominance of sensory neuropathy was earlier described in Sjogren's syndrome (SS), which might precede the presence of sicca symptoms. The mechanism of sensory neuropathy in SS is unknown. Therefore, the aim of this study was to determine the quantitative changes of the different neuropeptide containing nerve terminals and the immunocompetent cells in labial salivary glands of primary SS. DESIGN: Immunohisto- and immunocytochemical methods were used for the detection of immunoreactive (IR) elements and the data were compared with the healthy controls. RESULTS: All of the investigated IR nerve fibres were found in different quantity and localisation in both of control and SS glands. The density of them was changed variously in SS. The number of the substance P (SP), neuropeptide Y (NPY) (P < 0.05), galanin (GAL) IR nerve terminals was decreased, however, the number of vasoactive intestinal polypeptide (VIP) and tyrosine beta-hydroxylase (TH) IR nerve fibres (P < 0.05) was increased compared to the control. There were no IR immunocompetent cells in the control materials, however, a large number of them showed IR for SP (46.2%) and NPY (34.4%) in the SS. The IR was demonstrated mainly in the mast cells, plasma cells and some of the lymphocytes. CONCLUSIONS: These neuropeptides might have a role in the sensory neuropathy; they might activate nociceptive and sympathetic pathways. Some neuropeptides (SP, NPY) are endogenous in the immune system and produced in certain conditions, e.g. inflammation and chronic autoimmune disorders such as SS, so they might participate in the neuroimmunomodulation and contribute to the atrophy, apoptosis and necrosis. PMID- 15126136 TI - Modulation of voltage-dependent calcium channels by neurotransmitters and neuropeptides in parasympathetic submandibular ganglion neurons. AB - The control of saliva secretion is mainly under parasympathetic control, although there also could be a sympathetic component. Sympathetic nerves are held to have a limited action in secretion in submandibular glands because, on electrical stimulation, only a very small increase to the normal background, basal secretion occurs. Parasympathetic stimulation, on the other hand, caused a good flow of saliva with moderate secretion of acinar mucin, plus an extensive secretion of granules from the granular tubules. The submandibular ganglion (SMG) is a parasympathetic ganglion which receives inputs from preganglionic cholinergic neurons, and innervates the submandibular salivary gland to control saliva secretion. Neurotransmitters and neuropeptides acting via G-protein coupled receptors (GPCRs) change the electrical excitability of neurons. In these neurons, many neurotransmitters and neuropeptides modulate voltage-dependent calcium channels (VDCCs). The modulation is mediated by a family of GPCRs acting either directly through the membrane delimited G-proteins or through second messengers. However, the mechanism of modulation and the signal transduction pathway linked to an individual GPCRs depend on the animal species. This review reports how neurotransmitters and neuropeptides modulate VDCCs and how these modulatory actions are integrated in SMG systems. The action of neurotransmitters and neuropeptides on VDCCs may provide a mechanism for regulating SMG excitability and also provide a cellular mechanism of a variety of neuronal Ca(2+)-dependent processes. PMID- 15126137 TI - Bolus size and unilateral chewing cycle kinematics. AB - The purpose of this study was to determine how bolus size alters the human chewing cycle. This prospective within-subject design evaluated chewing cycles of 38 young adults between 20 and 38 years of age (21 males and 17 females). An optoelectric jaw tracking system was used to record movements of the chin during unilateral (right sided) chewing of four randomly ordered bolus sizes (1, 2, 4 and 8 g) of gum. Using each subject's 10 most representative cycles, multilevel statistical procedures were used to evaluate jaw kinematics. The results showed that bolus size has no consistent effect on opening, closing or total cycle duration. Cycle excursions increased significantly with increasing bolus size. With increasing bolus sizes, chewing cycle excursions along the three axes increased 52-115%. The greatest differences between bolus sizes occurred when the jaw was changing direction (i.e. passing from opening to closing and from working to balancing sides). However, the increases were proportionate and the shape of the chewing cycle was maintained. In order to maintain cycle duration while increasing excursive ranges, jaw velocities increased significantly, with the greatest differences occurring at approximately 70% of opening and 30% of closing. We conclude that humans adapt to larger bolus sizes by increasing chewing cycle perimeter and by increasing cycle speed, while maintaining cycle shape and duration. PMID- 15126138 TI - Correlation between intraoral pressures and tongue movements in the suckling pig. AB - The objective was to clarify the relationship between tongue movements during suckling and the pressures in different parts of the oral cavity. A modified teat allowed a miniature pressure transducer to be passed through into the mouth. Intraoral pressures were recorded in piglets suckling on the teat attached (1) to a non-vented bottle or (2) to an automated milk delivery system. The movements of the tongue, of the milk and the transducer position were recorded by cine radiography. In both modes of feeding, waves of elevation on the tongue moved in a pharyngeal direction and rose to contact the mid-posterior palate. Each wave corresponded to a jaw (suck) cycle in which milk was moved into and through the oral cavity. After each wave passed the transducer in the anterior part of the mouth, cyclical negative pressures were recorded. In bottle feeding, the intraoral pressure fluctuations (+/-2 mmHg) occurred against a background of a gradually developing negative pressure but, when feeding on the automatic delivery system, the same or smaller fluctuations occurred as changes from atmospheric pressure. Where the elevations contacted the mid-posterior palate in each cycle, a seal was formed (contact pressure >40 mmHg), so producing two functional antero-posterior compartments within the mouth; in these compartments pressures were generated independently. With the transducer in the valleculae, no general increase in pressure was recorded as milk accumulated there in each suck cycle but large positive pressures were recorded during the less frequent cycles when the vallecular space was emptied. PMID- 15126139 TI - Effects of growth factors on temporomandibular joint disc cells. AB - The effects of growth factors on cartilaginous tissues are well documented. An exception is the temporomandibular joint (TMJ) disc, where data for growth factor effects on proliferation and biosynthesis are very limited. The purpose of this study was to quantify proliferation of and synthesis by TMJ disc cells cultured in monolayer with either platelet derived growth factor-AB (PDGF), basic fibroblast growth factor (bFGF) or insulin-like growth factor-I (IGF), at either a low (10 ng/ml) or high (100 ng/ml) concentration. Proliferation was assessed with a DNA quantitation technique, collagen synthesis was measured via a hydroxyproline assay, and GAG synthesis was determined with a dimethylmethylene blue dye binding assay at 14 days. Overall, the most beneficial growth factor was bFGF, which was most potent in increasing proliferation and GAG synthesis, and also effective in promoting collagen synthesis. At the high concentration, bFGF resulted in 96% more cells than the control and 30 to 45% more cells than PDGF and IGF. PDGF and bFGF were the most potent upregulators of GAG synthesis, producing 2-3 times more GAG than the control. IGF had no significant effect on GAG production, although at its higher concentration it increased collagen production by 4.5 times over the control. Collagen synthesis was promoted by bFGF at its lower concentration, with levels 4.2 times higher than the control, whereas PDGF had no significant effect on collagen production. In general, higher concentrations increased proliferation, whereas lower concentrations favoured biosynthesis. PMID- 15126140 TI - Influence of growth hormone on the mandibular condylar cartilage of rats. AB - Growth hormone (GH) stimulates mandibular growth but its effect on the mandibular condylar cartilage is not well understood. OBJECTIVE: This study was designed to understand the influence of GH on mitotic activity and on chondrocytes maturation. The effect of GH on cartilage thickness was also determined. DESIGN: An animal model witt differences in GH status was determined by comparing mutant Lewis dwarf rats with reduced pituitary GH synthesis (dwarf), with normal rats and dwarf animals treated with GH. Six dwarf rats were injected with GH for 6 days, while other six normal rats and six dwarf rats composed other two groups. Mandibular condylar tissues were processed and stained for Herovici's stain and immunohistochemistry for proliferating cell nuclear antigen (PCNA) and alkaline phosphatase (ALP). Measurements of cartilage thickness as well as the numbers of immunopositive cells for each antibody were analysed by one-way analysis of variance. RESULTS: Cartilage thickness was significantly reduced in the dwarf animals treated with GH. PCNA expression was significant lower in the dwarf rats, but significantly increased when these animals were treated with GH. ALP expression was significant higher in the dwarf animals, while it was significantly reduced in the dwarf animals treated with GH. CONCLUSIONS: The results from this study showed that GH stimulates mitotic activity and delays cartilage cells maturation in the mandibular condyle. This effect at the cellular level may produce changes in the cartilage thickness. PMID- 15126141 TI - The expression of fibroblast growth factor receptor-3 in synovial osteochondromatosis of the temporomandibular joint. AB - Primary synovial osteochondromatosis (PSC) is a disease of unknown aetiology. It was reported recently that expression of fibroblast growth factor receptor-3 (FGFR-3) was observed specifically in PSC. We classified six cases of synovial osteochondromatosis (SC) of the temporomandibular joint (TMJ) into two types of SC, PSC (five cases) and secondary synovial osteochondromatosis (SSC) (one case), by means of clinical findings and haematoxylin and eosin stain. The five PSC cases were classified into three different phases according to Milgram's classification. Immunohistochemical staining of FGFR-3 was carried out for each SC case, along with specimens of internal derangement (ID) of the TMJ, and normal articular disc and synovial membrane. FGFR-3 was found in all three phases of PSC, but not in SSC, ID or normal TMJ. Moreover, in a comparison between cultured synovial cells of PSC (Phase III) and ID, reverse transcription-polymerase chain reaction revealed a stronger positive reaction in PSC. These results indicate that the synovial membrane in Phase III PSC can produce cartilage nodules, as in Phases I and II. PMID- 15126142 TI - Quantitative assessment of structural damage in eyes with localized visual field abnormalities. AB - PURPOSE: To evaluate the pattern of structural damage in the macula and peripapillary retinal nerve fiber layer (RNFL) using optical coherence tomography (OCT) and scanning laser polarimetry (SLP-VCC) in glaucomatous eyes with localized visual field defects. DESIGN: Prospective, cross-sectional analysis. METHODS: Complete examination, automated achromatic perimetry (AAP), Stratus OCT imaging (512 A-scans) of the peripapillary retina and macula, and SLP-VCC imaging of the peripapillary RNFL were performed. Thickness values in the retinal segments associated with the visual field defect (glaucomatous segments) were compared with corresponding segments across the horizontal raphe (nonglaucomatous segments) and age-matched normal controls. RESULTS: Forty eyes of 40 patients (20 normal, 20 glaucomatous) were enrolled (mean age, 71 +/- 10 years; range, 50 to 89). Mean RNFL thickness using SLP-VCC and OCT in the nonglaucomatous segments of glaucomatous eyes (54.0 +/- 9.7 microm, 64.7 +/- 19.0 microm) were significantly (P =.009, <0.0001) reduced compared with the thickness measurements in the corresponding segments of age-matched normal subjects (62.5 +/- 9.2 microm, 105.6 +/- 19.0 microm) respectively. No significant (P =.4) differences in the macular thickness measurements were observed between nonglaucomatous (239.0 +/- 19.4 microm) and normal segments (243.5 +/- 15.0 microm). Compared with age-matched controls, RNFL thickness in the nonglaucomatous segment was abnormal in 15 of 20 patients (75%) with SLP-VCC and in 18 of 20 patients (90%) with OCT. Macular thickness in the nonglaucomatous segment was abnormal in 11 of 20 patients (55%). CONCLUSIONS: Diffuse RNFL and retinal ganglion cell loss is present in eyes with localized visual field abnormalities. Detection of localized changes in macular thickness is limited by measurement overlap among normal and glaucomatous eyes. PMID- 15126143 TI - Topographically supported customized ablation for the management of decentered laser in situ keratomileusis. AB - PURPOSE: To evaluate the efficacy, predictability, and safety of topographically supported customized ablations (TOSCAs) for decentered ablations following laser in situ keratomileusis (LASIK). DESIGN: Prospective nonrandomized clinical trial. METHODS: Nine patients (11 eyes) with LASIK-induced decentered ablations underwent TOSCA following flap lifting. Topographically supported customized ablation was performed using a corneal topographer to obtain a customized ablation profile, combined with a flying spot laser. RESULTS: Mean follow-up was 9.22 +/- 2.82 months (range 6-12 months). No intra- or postoperative complications were observed. Manifest refraction (spherical equivalent) did not change significantly (pre-TOSCA: -0.14 +/- 1.58 diopters [range, -1.75 to +3.00 diopters] to +0.46 +/- 1.02 diopters [range, -1.00 to +1.75 diopters]; P =.76), whereas there was a statistically significant reduction in the refractive astigmatism (pre-TOSCA: -1.55 +/- 0.60 diopters [range, -3.00 to -0.75 diopters] to -0.70 +/- 0.56 diopters [range, -2.00 to -0.25 diopters]; P =.003). Mean uncorrected visual acuity improved significantly (P <.001) from 0.45 +/- 0.16 (range, 0.2-0.7) to 0.76 +/- 0.29 (range, 0.2-1.2) at last follow-up. Mean best corrected visual acuity improved from 0.74 +/- 0.22 (range, 0.4-1.0) to 0.95 +/- 0.20 (range, 0.6-1.2; P =.002). Eccentricity showed a statistically significant reduction after TOSCA treatment (pre-TOSCA: 1.59 +/- 0.46 mm [range, 0.88-2.23 mm]; post-TOSCA: 0.29 +/- 0.09 mm [range, 0.18-0.44 mm]; P <.001). CONCLUSION: In our small sample, enhancement LASIK procedures with TOSCA appear to improve uncorrected and best-corrected visual acuity as well as eccentricity in patients with LASIK-induced decentered ablation. PMID- 15126144 TI - Photodynamic therapy of subfoveal neovascular membrane in type 2A idiopathic juxtafoveolar retinal telangiectasis. AB - PURPOSE: To evaluate the effect of photodynamic therapy on subfoveal neovascular membrane related to type 2A idiopathic juxtafoveolar retinal telangiectasia. DESIGN: Interventional case series. METHODS: Retrospective review of four eyes of four patients who underwent photodynamic therapy for subfoveal neovascular membrane secondary to idiopathic juxtafoveolar retinal telangiectasia. Ocular photodynamic therapy with verteporfin was performed in all cases using standard protocols. Results are given in terms of final visual acuity and neovascular membrane activity based on clinical examination, fluorescein and indocyanin green angiography, and, in two cases, optical coherence tomography. RESULTS: Baseline visual acuity of 20/30 and 20/40 (x2) was maintained in three patients after one, two, and three sessions of photodynamic therapy respectively, and a follow-up of 23, 21, and 9 months. Leakage specific to the subfoveal neovascular membrane ceased on the fluorescein angiography. In the other patient, the final vision decreased from 20/50 to 20/200 after four sessions of photodynamic therapy and a follow-up of 14 months. Although there was still mild persistent leakage on the fluorescein angiography, neovascular membrane size was unchanged, and no subretinal fluid was demonstrated on optical coherence tomography. CONCLUSIONS: Data from this case series suggest that photodynamic therapy may be effective in managing subfoveal neovascular membrane associated with idiopathic juxtafoveolar retinal telangiectasia, which usually carries a poor visual prognosis. Prospective study is required to confirm the beneficial effect of this treatment. PMID- 15126145 TI - In vitro investigation of voriconazole susceptibility for keratitis and endophthalmitis fungal pathogens. AB - PURPOSE: To update the spectrum of ocular fungal isolates and investigate the in vitro efficacy of voriconazole and other antifungals. DESIGN: Experimental study. METHODS: Microbiology database was scanned and fungal isolates associated with keratitis (419) and endophthalmitis (122) were analyzed for classification and isolate frequency. The Sensititre YeastOne microdilution antifungal susceptibility test was used to evaluate susceptibility (MICs) of 34 common fungal pathogens against amphotericin B, fluconazole, ketoconazole, 5 flucytosine, itraconazole, and voriconazole. Ten of the test isolates were sent to a reference laboratory to validate the Sensititre results. RESULTS: Fusarium species remains the most frequent corneal fungal pathogen (60.1%). Colletotrichum species (4.1%) has emerged as the fifth most common mold in keratitis. Top yeast isolates from cornea included Candida albicans (52.3%) and Candida parapsilosis (37.3%). Half of the intraocular pathogens were Candida species. Paecilomyces (2.9%) and Philophora (1.9) were unusual pathogens. In vitro susceptibility profiles were voriconazole (100%), ketoconazole (82.4%), amphotericin (76.5%), itraconazole (67%), fluconazole (60%), and 5-FC (60%). Voriconazole MIC(90) were lowest for Candida species (0.016 microg/ml) and highest for Fusarium species (2 microg/ml). Reference laboratory MICs correlated 100% for yeast isolates (0.016 microg/ml) but were fourfold higher for Fusarium species (8 microg/ml). MIC(90) for Aspergillus species was 0.5 microg/ml. CONCLUSIONS: Candida, Fusarium, and Aspergillus species remain frequent fungal pathogens. Voriconazole may have a role in the therapeutic management of Candida and Aspergillus ocular infections. Clinical efficacy must determine the role for other fungal pathogens. Human use and animal models will determine its use in the clinical setting. PMID- 15126146 TI - Transient reduction in retinal function revealed by multifocal electroretinogram after photodynamic therapy. AB - PURPOSE: To evaluate the early changes in retinal function after photodynamic therapy (PDT) by multifocal electroretinogram (mfERG). DESIGN: Prospective interventional case series. METHODS: Seventeen eyes from 17 patients scheduled for standard PDT with verteporfin were prospectively recruited. Patients' diagnoses included choroidal neovascularization (CNV) secondary to age-related macular degeneration, idiopathic CNV, myopic CNV, polypoidal choroidal vasculopathy, and central serous chorioretinopathy. Serial mfERG recordings were performed before PDT, and at 4 days, 2 weeks, and 1 month after PDT. The first order kernel N1 and P1 mfERG response from the central 0 to 7 degrees and peripheral 7 to 25 degrees were grouped and analyzed. The mean response amplitudes and peak latencies of the mfERG recordings were compared longitudinally. RESULTS: There were statistically significant reductions in the mean N1 response amplitude for the central group at 4 days (P =.007) and 2 weeks after PDT (P =.024), in the mean P1 response amplitude for both the central (P =.006) and peripheral (P =.013) groups at 4 days, and for the central group at 2 weeks after PDT (P =.017). There were also statistically significant increases in the mean P1 response latencies at 4 days (P =.004) and at 2 weeks (P =.018) after PDT for the central group and at 4 days after PDT (P =.026) for the peripheral group. At 1 month after PDT, no significant differences in the N1 and P1 mean response amplitudes and peak latencies were observed compared with pre-PDT mfERG. CONCLUSIONS: Transient impairments in retinal function for as long as 2 weeks after PDT were noticed by reduction in response amplitudes and a delay in peak latencies of mfERG. These findings may explain the common adverse event of subjective visual disturbance early after PDT with normal findings in visual acuity and ophthalmoscopy. PMID- 15126147 TI - Evaluation of arteriovenous crossing sheathotomy for branch retinal vein occlusion by fluorescein videoangiography and image analysis. AB - PURPOSE: We quantitatively evaluated the effects of arteriovenous (A/V) crossing sheathotomy on retinal circulation in patients with branch retinal vein occlusion (BRVO) accompanied by macular edema. DESIGN: Interventional case series. METHODS: In 18 consecutive patients (18 eyes) with BRVO accompanied by macular edema who underwent A/V crossing sheathotomy between August 1999 and April 2002, changes in retinal circulation after the surgery were evaluated by fluorescein videoangiography with a scanning laser ophthalmoscope and by image analysis using dye dilution technique. At a venule distal to the responsible A/V crossing site and a normal venule, the circulation time (T50) from the beginning of filling to 50% filling of the peak intensity was calculated. The time difference (DeltaT50) between T50 at the point on the affected venule and that at the point on the normal venule, which represents the filling delay at the venule distal to the A/V crossing site, was compared between before and early after the surgery. RESULTS: The preoperative DeltaT50 was 1.36 +/- 1.15 seconds (mean +/- SD), and the postoperative DeltaT50 was 0.72 +/- 0.77 seconds (P =.035, paired t test). In 11 of the 18 eyes, DeltaT50 decreased by 20% or more after the surgery. In the other 7 eyes, DeltaT50 was unchanged or slightly increased after the surgery. CONCLUSIONS: Although a randomized controlled study is needed to confirm the effectiveness of A/V crossing sheathotomy on visual function, this technique could be effective for improving the delay in perfusion in the affected venule. PMID- 15126148 TI - Visual short-term effects of Viagra: double-blind study in healthy young subjects. AB - PURPOSE: To investigate short-term visual effects of a single 100-mg dose of Viagra (sildenafil citrate) in healthy men. DESIGN: Randomized, double-blind, placebo-controlled clinical trial of drug effects on normal volunteers conducted by a single center. METHODS: Twenty men, aged 20 to 40 years, were treated with either a placebo or 100 mg sildenafil. Visual function tests included electroretinogram (ERG) recordings, on-/off- and 3.3 Hz-flicker-ERG recordings, anomaloscope matches, and measurements of cone contrast sensitivities and transient tritanopia. RESULTS: Most visual tests did not differ between the sildenafil and placebo groups. However, statistically significant increases in sensitivity during transient tritanopia were observed as well as significant prolongations in the implicit times of scotopic a-wave, photopic b-wave, and 3.3 Hz-flicker a-wave and b-wave ERG recordings. The magnitude of the differences correlated with peak sidenafil plasma concentration. Although rod amplitudes of the ERG recordings tended to be higher and cone amplitudes lower in the sildenafil group after drug ingestion, the differences were nonsignificant. There were no reports of visual side effects, and all electrophysiologic and psychophysical measurements returned to the normal range within 24 hours. CONCLUSIONS: A single oral dose of 100-mg sildenafil given to healthy young men led to small but statistically significant transient changes of outer and inner retinal function, as detected by ERG and psychophysical methods. Although the acute effects were fully reversible within 24 hours, it would be worthwhile to compare them with those induced by other PDE5 and PDE6 inhibitors. PMID- 15126149 TI - Endothelial cell activation and hypercoagulability in ocular Behcet's disease. AB - PURPOSE: To investigate the presence of a hypercoagulable state and vascular endothelial dysfunction in patients with ocular Behcet's disease and relate the results to the activity of ocular and systemic involvement. DESIGN: Cross sectional laboratory and clinical study. METHODS: Prospective study of blood samples of 24 patients diagnosed with ocular Behcet's disease, which were analyzed for factor VIII, factor XI, von Willebrand factor antigen and ristocetin (vWF ag and risto), antithrombin III (ATIII), protein C and S, fibrinogen and activated protein C (APC) resistance. The results were compared with 40 healthy controls and analyzed for association with ocular and systemic clinical features. RESULTS: The mean values of factor VIII, factor XI, vWF ag, vWF risto, ATIII, and fibrinogen were significantly raised compared to healthy population (for all: P <.001). Most striking were factor VIII activity levels above 130% in 79% (19 of 24) of our patients. 67% (16 of 24) had levels of factor VIII above 150%, which correlates with a fivefold increase in risk of thrombosis. Other prothrombogenic factors were negative in all but 2 patients (1 protein C deficiency, 1 factor V Leiden mutation). Endothelial cell activation, measured by vWF activity, revealed elevated levels in 42% (10/24). Complete/incomplete Behcet's disease patients with present or previous macular edema had significantly higher FVIII levels than complete/incomplete Behcet's disease patients who had never shown any signs of macular edema (P =.04). Further correlations between the laboratory results and clinical symptoms were not found. CONCLUSIONS: We found a generalized hypercoagulable state with endothelial cell activation in ocular Behcet's disease, irrespectively of current ocular disease activity. PMID- 15126150 TI - Ultrasound biomicroscopic findings in aniridia. AB - PURPOSE: To describe the ultrasound biomicroscopic features of eyes with aniridia. DESIGN: Observational case series. METHODS: Nineteen eyes of 10 patients with aniridia (six males and four females) ranging in age from 3 months to 53 years (21.0 +/- 16.4, mean +/- SD), and 50 normal subjects (30 men and 20 women) ranging from 16 to 56 years (31.1 +/- 13.2) were evaluated. Ultrasound biomicroscopic findings were recorded in the 3-, 6-, 9-, and 12-o'clock directions. Adult patients (aged 16 years or older) with aniridia were compared with the age-matched controls. RESULTS: Ultrasound biomicroscopy (UBM) detected extremely tiny irises in all eyes with aniridia. The eyes with aniridia showed significantly smaller values than the controls in ciliary body length (4.49 +/- 0.63 versus 5.79 +/- 0.44 mm, P <.001, unpaired Student t test), ciliary body thickness (0.75 +/- 0.17 versus 1.24 +/- 0.22 mm, P <.001), iris root thickness (0.47 +/- 0.14 versus 0.61 +/- 0.07 mm, P <.001), scleral-ciliary process angle (31.7 +/- 3.26 versus 43.1 +/- 4.48 degree, P <.001), and anterior chamber depth (1.99 +/- 0.43 versus 2.94 +/- 0.34 mm, P <.001). In the aniridia eyes, there was a significantly positive correlation between iris thickness and ciliary body thickness (Pearson r = 0.829, P =.001). CONCLUSION: Ultrasound biomicroscopic imaging demonstrated that not only iris hypoplasia but also ciliary body hypoplasia exist in aniridia. Anterior inclination of the ciliary process was also found, which was thought to be at least partly responsible for the shallow anterior chamber. PMID- 15126151 TI - Frequency doubling technology perimetry abnormalities as predictors of glaucomatous visual field loss. AB - PURPOSE: To determine whether frequency doubling technology (FDT) perimetry results predict glaucomatous visual field defects, as assessed by standard automated perimetry (SAP), in a glaucoma suspect population. DESIGN: Longitudinal observational study. METHODS: The study included 105 eyes of 105 glaucoma suspect patients, with a mean follow-up time of 41 +/- 17 months. Glaucoma suspects had either intraocular pressure (IOP) higher than or equal to 23 mm Hg or glaucomatous optic neuropathy by stereophotograph assessment. All patients had normal SAP visual fields at baseline. A baseline FDT test was performed within 3 months of the normal SAP examination. Several baseline FDT parameters and other variables (age, gender, IOP, central corneal thickness, SAP visual field indices, and stereophotograph assessment) were investigated by univariate and multivariate Cox proportional hazards models to obtain hazard ratios (HR) and identify factors that predicted which patients had SAP glaucomatous visual field loss during follow-up. RESULTS: Seventeen patients (16%) developed repeatable SAP visual field abnormality during follow-up. An abnormal FDT examination at baseline predicted the development of SAP visual field conversion in both univariate (HR = 3.17; 95% confidence interval [CI] = 1.22-8.25; P =.018) and multivariate models (Adjusted HR = 3.68; 95% CI = 1.06-12.8; P =.04). The analysis of FDT examinations during follow-up revealed that in 59% of converters the FDT abnormalities preceded SAP visual field loss by as much as 4 years. Also, the initial development of glaucomatous visual field loss as measured by SAP occurred in regions that had previously demonstrated abnormalities on FDT testing. CONCLUSION: Functional abnormalities detected by FDT perimetry were predictive of the future onset and location of SAP visual field loss among glaucoma suspect patients. PMID- 15126152 TI - Clinical characteristics of acute HSV-2 retinal necrosis. AB - PURPOSE: To report the clinical features and evaluate the visual outcome of eleven cases of herpes simplex virus-2 (HSV-2) related acute retinal necrosis syndrome (ARN). DESIGN: Retrospective interventional case series. METHODS: Twelve eyes of eleven patients from two European centers, diagnosed with HSV-2 related acute retinal necrosis syndrome were retrospectively reviewed. Herpes simplex virus-2 DNA was detected by polymerase chain reaction in intraocular fluids (aqueous and/or vitreous). Findings at initial examination, clinical evolution with antiviral therapy, complications and final visual acuity were evaluated. RESULTS: Herpes simplex virus-2 DNA was detected in all cases. No sample was positive for more than one virus. The mean age of disease in the first eye was 36 years (ranged from 10 to 57 years). Five patients were women and six were men. All patients were immunocompetent. Previous medical history included neonatal herpes (n = 1), previous ARN (n = 3), trauma (n = 1) and systemic corticosteroid administration before occurrence of ARN (n = 3). Preexisting pigmented chorioretinal scars were found in three cases. Patients were treated with high dose intravenous acyclovir or foscarnet +/- intravitreal ganciclovir +/- interferon. The mean follow-up was 14.5 months (from 5 to 22 months). At the end of the follow-up period, five eyes (41.7%) showed improvement of visual acuity of two or more lines. Final visual acuity was 20/60 or better in four eyes (33.3%), 20/400 or better in four eyes (33.3%) and less than 20/400 in four eyes. CONCLUSION: History of neonatal herpes, triggering events such as neurosurgery, periocular trauma, high-dose corticosteroids, and chorioretinal scars suggest that HSV-2 retinitis reflects reactivation of HSV-2 infection. PMID- 15126153 TI - Limited macular translocation compared with photodynamic therapy in the management of subfoveal choroidal neovascularization in age-related macular degeneration. AB - PURPOSE: To compare the visual outcome of macular translocation (MT) versus photodynamic therapy (PDT) for subfoveal predominantly classic neovascularization in age-related macular degeneration (AMD). DESIGN: Nonrandomized clinical trial. METHODS: Retrospective review of 65 consecutive patients with subfoveal neovascularization due to AMD. The follow-up was at least 6 months. Main outcome criteria were final best corrected visual acuity and the gain in visual acuity. RESULTS: A total of 29 eyes were treated with PDT with verteporfin, and 36 underwent MT with chorioscleral infolding. Both groups were similar for age, refraction, and lesion size. The initial visual acuity was lower in the MT group than in the PDT group (20/200 versus 20/100). Mean follow-up was 11 months for the PDT group and 14 months for the MT group. The mean displacement of the fovea after translocation was 1,274 microm (range, 250 to 1,900 microm). Mean number of retreatment by PDT was 2.5. At 1 year, both groups had the same final visual acuity (20/200), but the improvement was more favorable in the MT group (gain of 0.7 line in the MT group versus loss of 3.4 lines in the PDT group, P =.007). One eye in the PDT group (4.3%) had a gain of 3 lines or more versus eight eyes (38%) in the MT group; the lesion size was larger in the PDT group than in MT group (P =.036). CONCLUSION: In this retrospective study, MT seemed to allow a better preservation of visual acuity than PDT in subfoveal neovascularization due to AMD. Further larger and controlled studies are required. PMID- 15126154 TI - Ocular findings in leprosy patients in Nepal in the era of multidrug therapy. AB - PURPOSE: Detailed ophthalmic evaluation was performed to determine the prevalence of ocular complications among leprosy patients on multidrug therapy and those released from multidrug treatment. DESIGN: Observational case series. METHODS: Leprosy patients at Tribhuvan University Teaching Hospital from April 1, 2001, through September 30, 2002, underwent detailed ophthalmic evaluation including slit-lamp biomicroscopy, dilated funduscopy, and applanation tonometry. RESULTS: We evaluated 58 leprosy patients. A majority (72%) was receiving treatment for multibacillary leprosy; 14% belonged to posttreatment multibacillary and paucibacillary groups. Ocular involvement was found in 57% of patients. In the multibacillary group, 55% had ocular involvement, which was more than double that found in the paucibacillary group (25%), although this finding was not statistically significant (P =.187). Among patients with ocular complications, 48% had visual disability and another 45% had threatened vision; 9% met World Health Organization guidelines for blindness. Uveitis and its complications were the predominant causes of visual disability (88%). CONCLUSION: Ocular complications and visual disability are high among leprosy patients in Nepal even after completing multidrug therapy. PMID- 15126155 TI - Survival, anatomic, and functional long-term results in choroidal and ciliary body melanoma after ruthenium brachytherapy (15 years' experience with beta rays). AB - PURPOSE: To analyze survival, radiation-related complications, local recurrence, enucleation, and visual acuity after ruthenium-106 irradiation of malignant uveal melanoma. DESIGN: Retrospective study. METHODS: A total of 213 patients treated with ruthenium-106 brachytherapy between 1983 and 1995. End point rates were estimated by life tables, and prognostic factors by Cox proportional hazards regression. Main outcome measures were survival rate, radiation-related maculopathy, radiation-related vascular occlusion, local tumor recurrence, enucleation, and visual acuity (<20/100 at 3 years). RESULTS: At 5 and 10 years, survival was 82% (standard error [SE] 2.7%) and 72% (SE 3.4%), local recurrence 21.7% (SE 3.0%) and 24.3% (SE 3.2%), and enucleation 18.0% (SE 2.7%) and 19.2% (SE 2.8%), respectively. Sixty-one patients showed maculopathy (29%), 36 retinal vascular occlusion (17%), in 33 local recurrence (16%), and 38 enucleation (18%). Age and large tumor diameter were independently associated with survival (P <.0001 and P <.0075, respectively). Age below 40 and melanoma located posteriorly were significant risk factors for maculopathy (P <.0085 and P <.0004, respectively) and vascular occlusion (P <.0415 and P <.0114, respectively). Diameter and Bruch membrane rupture were significant predictors (P <.0032 and P <.0390, respectively) of local recurrence. Visual acuity <20/100 was observed in only 26 of 97 (27%) cases of anterior but 34 of 42 (81%) of posterior tumor (P <.001). CONCLUSION: Although percentage tumor recurrence was high, survival was comparable to series using other treatments. Radiation-related complication rates were acceptable, especially for anterior tumors. Ruthenium therapy can, therefore, be recommended for small and medium-sized tumors with anterior location. PMID- 15126156 TI - Use of rotational sutures for limited retinal translocation: a new technique for superior limited macular translocation. AB - PURPOSE: To report a modified surgical technique for retinal translocation in eyes with subfoveal choroidal neovascularization. DESIGN: Experimental animal study. METHODS: Nine pigmented rabbits were used consecutively to apply this technique. Placement of inferotemporal scleral imbrication sutures was followed by vitrectomy with posterior hyaloid separation. Balanced saline solution (BSS) was injected subretinally with a 30G needle or with a 39G hydrodissection cannula and viscous fluid injector to detach one retinal quadrant. Under low intraocular pressure, the imbrication sutures were tied, the sclerotomy sites were closed, and intravitreal air tamponade was injected. Rotation sutures were passed and the eye globe was rotated approximately 90 degrees counterclockwise. The rotation sutures were removed after 24 hours. Retinal photographs were taken and fundus examination was performed on postoperative days 1, 2 and 7. The animals were sacrificed after 7 to 10 days for postmortem macroscopic examination. RESULTS: The entire procedure was performed in nine eyes of nine rabbits. In eight eyes, translocation could be seen on the first postoperative day after removal of the rotation sutures. The average amount of translocation was 667 microm (range: 500 800 microm) in a nasal to inferonasal direction. Vitreous hemorrhage occurred at the end of surgery in one eye due to hypotony. Iatrogenic small retinal breaks occurred in 2 eyes but did not prevent completion of the procedure. There was only a temporary hyperemia of the eyelids and conjunctiva. CONCLUSION: Limited retinal translocation using rotational sutures provided a predictable amount of translocation in the planned direction. This technique is expected to be useful for superior macular translocation in humans. PMID- 15126157 TI - Clinical features associated with lesions other than pituitary adenoma in patients with an optic chiasmal syndrome. AB - PURPOSE: Pituitary adenomas are the most common cause of an optic chiasmal syndrome, and treatment of these lesions is considerably different from the treatment of most of the other lesions in this region. Although the diagnosis of a pituitary adenoma is usually inferred from the results of neuroimaging, lesions other than pituitary adenomas can have an appearance that suggests an adenoma. The objective of our study was to determine whether there are clinical findings that suggest a lesion producing a chiasmal syndrome is something other than a pituitary adenoma. DESIGN: Retrospective, case-controlled, analysis of medical record data. METHODS: The records of the Neuro-Ophthalmology Unit of the Wilmer Eye Institute were searched for patients with a chiasmal syndrome who had been evaluated before treatment and for whom pathologic or laboratory confirmation of the etiology was available. Presenting clinical features of these patients were recorded, and analyses with both a single variable and multiple variables were performed to determine whether there were any features that could identify with a high degree of probability the etiology of the lesion producing the syndrome. RESULTS: The search revealed 149 patients who met the inclusion criteria, including 90 patients with pituitary adenomas and 59 patients with other lesions. Variables that were highly suggestive of an etiology other than pituitary adenoma included symptomatic visual loss, younger age, unilateral optic disk pallor, a relative afferent pupillary defect, and an absolute or a complete visual field defect or one was greater inferiorly than superiorly. CONCLUSION: Although no single clinical feature can be used to determine the specific nature of a lesion that produces an optic chiasmal syndrome, certain features are highly suggestive of an etiology other than pituitary adenoma. When these features are present, the likelihood that a suprasellar lesion is a pituitary adenoma is much lower, regardless of the appearance on neuroimaging. PMID- 15126158 TI - Immunohistopathologic evaluation of choroidal neovascular membranes following verteporfin-photodynamic therapy. AB - PURPOSE: To evaluate the vascularization and proliferative activity in choroidal neovascular membranes due to age-related macular degeneration after verteporfin photodynamic therapy and submacular removal. DESIGN: Interventional case series. METHODS: In a retrospective review of seven patients who underwent removal of subfoveal classic choroidal neovascular membranes after treatment with photodynamic therapy 3 to 146 days earlier, membranes were stained for CD 34, CD 105, and Ki-67 and correlated with clinical pictures and fluorescein angiography. RESULTS: Fluorescein angiography performed on the day of surgery disclosed nonperfusion of the treated area 3 days after photodynamic therapy, but perfusion and leakage were seen at greater post-photodynamic therapy intervals. Membranes excised 3 days after photodynamic therapy showed CD34 and CD105 positive, mostly occluded vessels. The endothelial cells appeared damaged. Ki-67 activity was low. In membranes excised 34 to 146 days after photodynamic therapy, all vessels appeared patent and were lined by healthy endothelial cells with strong expression of CD34 and CD105. Ki-67 expression was elevated after 34 days but decreased thereafter. CONCLUSION: Photodynamic therapy did not cause a general or complete occlusion of vessels within the choroidal neovascular membranes, as suggested by fluorescein angiography 3 days postintervention, but the endothelial cells appeared to be severely damaged. Proliferative activity within these specimens was reduced. At longer intervals after photodynamic therapy, the fibrovascular tissue seemed to recover; perfusion, hyperfluorescence, and leakage of the choroidal neovascular membranes could be detected by fluorescein angiography. The clinical appearance showed a correlation with the immunohistologic characteristics of an increased proliferative activity and patent vascularization. PMID- 15126159 TI - Comparison of color to fluorescein angiographic images from patients with early adult onset grouped drusen suggests drusen substructure. AB - PURPOSE: Recent structural and histochemical data from our laboratory indicate that human drusen often possess distinct, structural core domains. A similar, corelike structure is frequently noted clinically in some drusen-related conditions. To assess the nature of the corelike structures observed clinically, we evaluated color photographs and fluorescein angiograms from patients with early-adult onset grouped drusen (EAOGD). DESIGN: Retrospective case series. METHODS: The areas of the six largest drusen and hyperfluorescent lesions for eight patients confirmed to have EAOGD were estimated by two masked examiners (RRG, JCF). Data were normalized using a logarithmic transformation and evaluated using a mixed model analysis to account for intragrader, interobserver and intrasubject effects. RESULTS: The average color to hyperfluorescence drusen area ratio was 2.29 (SE, 0.06). The area of drusen viewed in color photographs significantly exceeds that of the corresponding area of hyperfluorescence (P <.0001). The average drusen area (geometric mean) was 0.014 mm(2) (SE, 0.001 mm(2)) and the average hyperfluorescent lesion area was 0.006 mm(2) (SE, 0.0005 mm(2)). The areas of the hyperfluorescent lesions do not vary among the capillary, venous, and washout angiographic phases. The lesions exceed choroidal intensity and demonstrate prominent, uniformly hyperintense fluorescence throughout all but the choroidal phases of the angiogram. CONCLUSIONS: These data show that large visible drusen in EAOGD are concentric to regions of hyperfluorescence, suggesting that drusen may possess clinically detectable, substructural domains. Ongoing studies seek to determine whether clinical evidence of drusen substructure may exist in other drusen-associated disorders, such as in age-related macular degeneration. PMID- 15126160 TI - Indocyanine green induces apoptosis in human retinal pigment epithelial cells. AB - PURPOSE: To examine whether indocyanine green (ICG) dye induces apoptosis in human retinal pigment epithelial (RPE) cells. DESIGN: Laboratory investigation. METHODS: Pure cultures of human RPE cells were isolated. Retinal pigment epithelial cells were incubated with different concentrations of ICG dye (1 mg/ml, 5 mg/ml, or 20 mg/ml) for 30 minutes. The rate of RPE cell apoptosis was assessed with Annexin V-FITC staining and propidium iodide (PI) by flow cytometry. RESULTS: Retinal pigment epithelial cells maintained their monolayer morphology after incubation with ICG dye. However, ICG induced a statistically significant amount of apoptosis in RPE cells at all the concentrations (1 mg/ml, 5 mg/ml, and 20 mg/ml) after 30 minutes of incubation (P <.05). The solvent solution alone (without the ICG dye) did not induce any significant apoptosis in RPE cells, when compared with culture medium. CONCLUSIONS: The incubation of RPE cells with ICG dye increased the number of apoptotic RPE cells in vitro. Our findings indicate that the decision over the intravitreal application of ICG dye needs to be made with caution. PMID- 15126161 TI - Computerized kinetic perimetry detects tubular visual fields in patients with functional visual loss. AB - PURPOSE: To report a novel test that uses computerized kinetic perimetry to detect tubular visual fields in patients with functional visual loss. DESIGN: Observational case series. METHODS: Ten patients who demonstrated generalized visual field constriction on prior examinations underwent testing combining computerized kinetic perimetry with a reversed Galilean telescope, used for optical visual field expansion. RESULTS: Six patients with physiologic visual field constriction demonstrated expansion of their visual fields with the use of the reversed telescope. Four patients with functional visual loss responded to the reversed telescope by demonstrating tubular visual fields, with responses equal to or smaller than their demonstrated visual fields delineated without the telescope. CONCLUSIONS: This test uses a computerized kinetic examination to reduce the subjective nature of tangent screen testing for tubular visual fields and to provide a computerized recording of visual fields. This method is a novel and easy-to-use technique to demonstrate functional visual loss. PMID- 15126162 TI - Decrease of pigment epithelium-derived factor in aqueous humor with increasing age. AB - PURPOSE: To determine whether the level of pigment epithelium-derived factor (PEDF) in the aqueous humor is altered with age. DESIGN: Observational case series. METHODS: The PEDF concentration in the aqueous humor was measured by enzyme-linked immunosorbent assay in 152 eyes of 121 patients who underwent cataract surgery. RESULTS: The mean aqueous level of PEDF was 0.86 +/- 0.04 microg/ml (mean +/- standard error, 70.7 +/- 1.0 years). The PEDF levels decreased with increasing age, and the decrease was significantly correlated with age (Pearson product moment correlation coefficient, r = - 0.22, P =.006). The mean PEDF level in the men (0.76 +/- 0.06 microg/ml, 53 eyes, 71.5 +/- 2.0 years) was significantly lower than that in women (0.91 +/- 0.04 microg/ml, 99 eyes, 70.2 +/- 1.1 year, P =.03). CONCLUSIONS: The negative correlation of PEDF level and age should be considered in age-related eye diseases, especially those associated with angiogenesis. PMID- 15126163 TI - Retinal protection using a viscoadaptive viscoelastic agent during removal of a luxated crystalline lens by intravitreal phacoemulsification. AB - PURPOSE: To report a method of retinal protection using a viscoadaptive viscoelastic agent during removal of a luxated crystalline lens by intravitreal phacoemulsification. DESIGN: Interventional case report. METHODS: A 65-year-old man presented with a totally luxated crystalline lens in the vitreous cavity of the left eye. After total vitrectomy was performed, a viscoadaptive viscoelastic agent was applied over the retina and optic disk. Then the luxated lens was removed by intravitreal phacoemulsification. The lens dropped several times during the surgery but the thick viscoadaptive viscoelastic agent protected the posterior retina. RESULTS: In two cases treated this way, no complications occurred at six months after surgery. CONCLUSION: This method may be useful for protecting the retina from damage by a luxated crystalline lens and is less costly than perfluorocarbon liquid. PMID- 15126164 TI - Predominant loss of the photopic negative response in central retinal artery occlusion. AB - PURPOSE: To determine how the photopic negative response (PhNR) is affected in central retinal artery occlusion (CRAO). DESIGN: Observational case series. METHODS: Seven patients with unilateral CRAO were included. Full-field scotopic and photopic electroretinograms (ERGs) including the PhNR were recorded. Each ERG amplitude in the affected eye was expressed as a percentage of amplitude of the corresponding wave in the unaffected eye. RESULTS: Mean of the PhNR amplitude was reduced to 12.3 +/- 11.7% of that of unaffected eyes whereas the cone b-wave amplitude was attenuated to only 73.4 +/- 30.4%. This reduction of the PhNR amplitude was more significant than that of other waves including the rod b-wave, maximum a-wave and b-wave, cone a-wave and b-wave, and 30 Hz flicker ERG (P <.005). CONCLUSIONS: The PhNR was severely affected in CRAO despite relative preservation of the cone b-wave, implicating massive loss of ganglion cells and their axons. PMID- 15126165 TI - Acute acquired toxoplasma retinitis may present similarly to unilateral acute idiopathic maculopathy. AB - PURPOSE: To describe a patient with acquired toxoplasma retinitis that shared similarities with unilateral acute idiopathic maculopathy. DESIGN: Interventional case report. METHODS: Interventional case report. RESULTS: A 39-year-old woman presented with sudden unilateral vision loss in the right eye after a flu-like illness. She had retinitis involving the fovea, subretinal fluid, thickening of the retinal pigment epithelium, and a few vitreous cells; the retinitis resolved with antitoxoplasma antibiotic treatment. CONCLUSIONS: Acute acquired toxoplasma retinitis may present similarly to unilateral acute idiopathic maculopathy. PMID- 15126166 TI - Panophthalmitis due to clostridium septicum. AB - PURPOSE: To describe patient survival in a rare case of endogenous Clostridium septicum sepsis with panophthalmitis. DESIGN: Observational case report. METHODS: Both eyes of a female patient were examined in a hospital setting. RESULTS: A 68 year-old woman had right orbital pain, proptosis, panophthalmitis, mental confusion and fever for 2 days. Blood cultures were significant for Clostridium septicum. The patient did not improve after treatment with intravenous broad spectrum antibiotics and the right eye was enucleated. The patient survived the acute infection and extensive systemic evaluation revealed an undiagnosed colon carcinoma that may have been responsible for colonization and vascular dissemination of Clostridium septicum. CONCLUSIONS: Clostridium septicum panophthalmitis and sepsis can be the presenting sign in patients with unsuspected malignancies, particularly colon cancer. Patients can survive the infection with aggressive therapy with systemic antibiotics combined with removal of the infected tissue. PMID- 15126167 TI - High-frequency ultrasound of anterior segment retinoblastoma. AB - PURPOSE: To evaluate anterior segment retinoblastoma with high-frequency ultrasound. DESIGN: Interventional case report. METHODS: An 11-year-old boy with a total retinal detachment and multiple white anterior chamber nodules was examined with high-frequency ultrasound before enucleation of the globe. RESULTS: Multiple solid and cystic nodular tumors were found in the anterior chamber angle. With high-frequency ultrasound, tumor was discovered posterior to the iris, over the ciliary processes, and on the lens capsule. No calcifications were noted. Other findings included angle closure related to iris neovascularization and uveal thickening. CONCLUSION: High-frequency ultrasound can be used to evaluate retinoblastoma extension into the anterior segment. These images may be used for comparison in future cases of retinoblastoma with anterior chamber invasion. PMID- 15126168 TI - Histologic study of retinitis pigmentosa due to a mutation in the RP13 gene (PRPC8): comparison with rhodopsin Pro23His, Cys110Arg, and Glu181Lys. AB - PURPOSE: To evaluate the retina in autopsy eyes from patients over age 60 with autosomal dominant retinitis pigmentosa and a mutation in the RP13 gene (designated as PRPC8, Arg2310Gly), rhodopsin Pro23His, rhodopsin Cys110Arg, or rhodopsin Glu181Lys. DESIGN: Histologic study of the retina. METHODS: All eyes were prepared for electron microscopy within 12 hours after death. RESULTS: All eyes showed loss of rod photoreceptors. Remaining cones showed perinuclear membranous swirls, inclusion bodies in the inner segments, and shortened or absent outer segments despite causation by various gene defects. CONCLUSION: The comparable histologic findings in these four cases suggest a final common pathway leading to photoreceptor cell death in these dominant forms of retinitis pigmentosa. PMID- 15126169 TI - Acute myeloid leukemia presenting as bilateral proptosis from diffuse extraocular muscle infiltration. AB - PURPOSE: To report a case of acute myeloid leukemia with bilateral proptosis as the sole presenting sign. DESIGN: Observational case report. METHODS: A patient with bilateral proptosis was seen in consultation by pediatric ophthalmology. RESULTS: Complete blood count, computerized tomography, and bone marrow biopsy confirmed the diagnosis of acute myeloid leukemia, with the proptosis due to diffuse infiltration of all extraocular muscles. CONCLUSION: In a child with the sudden onset of proptosis without any other systemic findings, the diagnosis of acute leukemia must be considered. PMID- 15126170 TI - Limbal stem cell deficiency following topical mitomycin C treatment of conjunctival-corneal intraepithelial neoplasia. AB - PURPOSE: To report a case of conjunctival-corneal intraepithelial neoplasia (CCIN) in an elderly African American patient treated with topical mitomycin C and the subsequent complication of limbal stem cell deficiency. DESIGN: Interventional case report. METHODS: A 92-year-old African American woman was diagnosed with CCIN in the right eye. Following incisional biopsy, the patient received five 1-week courses of 0.04% mitomycin C and was followed over a period of 10 months. RESULTS: The CCIN regressed completely following mitomycin C therapy. Three months later, the patient developed recurrent nonhealing epithelial defects in the right cornea. CONCLUSIONS: Conjunctival-corneal intraepithelial neoplasia may occur in the African American population. Although MMC is effective in eradicating CCIN, a limbal stem cell deficiency may complicate the treatment. PMID- 15126171 TI - SANDO: another presentation of mitochondrial disease. AB - PURPOSE: To describe a unique mitochondrial syndrome that may present with the combination of ocular manifestations, sensory symptoms, and speech dysfunction. DESIGN: Case report. METHODS: Case report and review of the literature. RESULTS: A 43-year-old man presented with a Sensory Ataxic Neuropathy, Dysarthria, and Ophthalmoparesis (SANDO). Nerve conduction studies showed a sensory ganglionopathy. Skeletal muscle biopsy revealed ragged red fibers, and polymerase chain reaction analysis of the tissue demonstrated multiple mitochondrial DNA deletions. CONCLUSION: Mitochondrial disease should be considered in cases of external ophthalmoplegia, especially if a sensory ganglionopathy and dysarthria are present. PMID- 15126172 TI - Clinical and anatomical reversal of long-term hypotony maculopathy. AB - PURPOSE: To report a case of clinical and anatomical reversal of long-term hypotony maculopathy after filtration surgery. DESIGN: Interventional/observational case report. METHODS: Case report and review of a 59 year-old woman who underwent surgical bleb revision for hypotony maculopathy. RESULTS: Over a 5-year period, progressive vision loss to 20/200 developed in the patient's right eye, with an intraocular pressure in the single digits. She underwent bleb excision, scleral patch graft, and conjunctival advancement. Five weeks later, her visual acuity had dramatically improved to 20/25 with an intraocular pressure of 16 mm Hg. The retinal thickness decreased from 423 microm to 211 microm based on ocular coherence tomography. CONCLUSIONS: This case demonstrates that long-term hypotony maculopathy is reversible both functionally and structurally. PMID- 15126173 TI - Vitreoretinal surgery with slit-lamp illumination combined with a wide-angle viewing contact lens. AB - PURPOSE: To evaluate the feasibility of illuminating the fundus by combining a wide-angle-viewing contact lens and slit lamp attached to a surgical microscope during retinal reattachment surgery and vitrectomy. DESIGN: An interventional study. METHODS: We combined slit-lamp illumination and a wide-angle-viewing contact lens to visualize the fundus during retinal reattachment surgery and vitrectomy. RESULTS: We clearly observed the fundus using this combination approach during conventional retinal reattachment surgery, which we completed without using an indirect ophthalmoscope. With slit-lamp illumination, the wide angle-viewing contact lens provided a wider area of illumination than a planoconcave contact lens during vitrectomy in addition to an area of illumination as wide as that obtained using a light pipe. CONCLUSIONS: Combining slit-lamp illumination and a wide-angle-viewing contact lens seems to provide a useful alternative method for viewing the fundus during vitreoretinal surgery. PMID- 15126174 TI - Autofluorescence as a novel approach to diagnosing macular holes. AB - PURPOSE: To report the capability of autofluorescence as a novel approach to diagnosing macular holes. DESIGN: Observational case report. METHODS: In an institutional study, autofluorescence photos and optical coherence tomography scans of two patients who had surgery for macular holes were reviewed. RESULTS: Macular hole cases had a strong subfoveal autofluorescence signal, which disappeared after surgical closure of the macular hole. CONCLUSIONS: Autofluorescence is a novel approach to demonstrating a macular hole. This technique can also be used to confirm postoperative outcome after macular hole repair. A lack of autofluorescence corresponds to macular hole closure. PMID- 15126175 TI - Optical coherence tomography to detect macular edema in the presence of asteroid hyalosis. AB - PURPOSE: To propose the use of optical coherence tomography as an effective diagnostic tool for identifying macular edema in patients with asteroid hyalosis obscuring the fundus view. DESIGN: Case report. METHODS: Review of clinical chart and images. SETTING: Private retina practice. PATIENT: One patient diagnosed with diabetic retinopathy and asteroid hyalosis, who was experiencing decreased visual acuity and whose fundus view was inadequate for diagnosis using customary techniques. RESULTS: Usual methods of diagnosis were ineffective until OCT obtained a clear image of the fundus and subsequent macular thickening with vitreomacular adhesion. CONCLUSIONS: Optical coherence tomography is an effective diagnostic tool for discovering macular edema in cases of dense asteroid hyalosis where traditional methods fail to obtain a clear image of the fundus. PMID- 15126176 TI - Is adjustment of National Eye Institute Visual Function Questionnaire scores for general health necessary in randomized trials? AB - PURPOSE: To assess whether treatment comparison of National Eye Institute Visual Function Questionnaire (NEI-VFQ) scores in a clinical trial is influenced by general health to warrant adjusting for it. DESIGN: Two randomized pilot trials. METHODS: Patients enrolled in two randomized pilot trials of submacular surgery versus observation for choroidal neovascularization had quality of life interviews (NEI-VFQ and the Short Form-36 Health Survey) 24 months after enrollment. Information on comorbidities was collected through chart reviews. Data from 120 patients were analyzed using linear regression methods. RESULTS: Adjustment for comorbidities did not change the magnitude of the treatment effect on NEI-VFQ scores. However, adjustment for Short Form-36 physical and mental component summaries produced changes in the estimated treatment effect when NEI VFQ scores were compared. CONCLUSIONS: Adjustment of NEI-VFQ scores for general health may be advisable. The Short Form-36 summary scores may be appropriate for this purpose. PMID- 15126177 TI - Best's vitelliform macular dystrophy with pseudohypopyon: an optical coherence tomography study. AB - PURPOSE: To report optical coherence tomography (OCT) changes in Best's vitelliform macular dystrophy (BVMD) with pseudohypopyon. DESIGN: Observational case report. METHODS: Both eyes of a patient with BVMD showing pseudohypopyon were examined with OCT. RESULTS: OCT demonstrated the presence of serous retinal elevation with normal appearing retinal pigment epithelium (RPE) superiorly, and broadening of the outer-retina-choroid-complex signal under retinal elevation, inferiorly. CONCLUSIONS: OCT findings in our study suggest the accumulation of material under neurosensory retina in BVMD with pseudohypopyon. PMID- 15126178 TI - Topical prostaglandin F(2alpha) analog induced poliosis. AB - PURPOSE: To report poliosis as a side effect associated with topical prostaglandin F(2alpha) (PGF(2alpha)) analogs. METHOD: Case series. RESULTS: Seven patients treated with different topical PGF(2alpha) analogs for primary open angle glaucoma developed bilateral poliosis, either alone or in combination with other adverse effects of PGF(2alpha) analog therapy. CONCLUSION: Poliosis is a possible adverse effect of topical PGF(2alpha) analog therapy which is previously unreported. Topical PGF(2alpha) analog therapy should be included in the differential diagnosis of patients with poliosis. PMID- 15126179 TI - Cystoid macular edema in a low-risk patient after switching from latanoprost to bimatoprost. AB - PURPOSE: To report a case of angiographically documented cystoid macula edema occurring after switching a pseudophakic patient from latanoprost to bimatoprost. DESIGN: Observational case report. METHODS: A 68-year-old man developed intense conjunctival hyperemia and cystoid macula edema after switching from latanoprost to bimatoprost 9 months after cataract surgery in an eye at low-risk for this cystoid macular edema. RESULTS: Bimatoprost was discontinued and diclofenac initiated. After 2 months, visual acuity and ocular hyperemia returned to baseline levels. Fundus examination revealed resolution of cystoid macula edema. CONCLUSION: It is possible that pseudophakic eyes that develop intense conjunctival hyperemia associated with ocular hypotensive lipids might be at higher risk for developing cystoid macula edema. PMID- 15126180 TI - Metastatic mesothelioma presenting with proptosis. AB - PURPOSE: To describe the clinical presentation and histologic findings in a patient with metastatic mesothelioma presenting to the ophthalmologist with nonaxial proptosis. DESIGN: Case report. METHODS: A 55-year-old man presented with a short history of progressive ocular discomfort and vertical diplopia. Clinical examination identified nonaxial proptosis. Subsequent computed tomography showed a large extraconal mass consistent with a malignant process. Three months earlier the patient had been diagnosed with pleural mesothelioma. Unfortunately, he died 3 months after his ophthalmic presentation. Postmortem examination confirmed metastatic mesothelioma in the orbital roof that was histologically identical to the primary pleural malignancy. CONCLUSION: Pleural mesothelioma can metastasize to the orbit, causing proptosis. PMID- 15126181 TI - Correcting the corneal power measurements for intraocular lens power calculations after myopic laser in situ keratomileusis. PMID- 15126183 TI - Radial optic neurotomy in retinal vein occlusion. PMID- 15126185 TI - New challenges and opportunities in health and development in Africa. PMID- 15126186 TI - Vitamin A deficiency among Moroccan women and children. AB - BACKGROUND: Vitamin A deficiency (VAD), attributable to an inadequate food intake, constitutes a problem in developing countries. The determination of plasma vitamin A (VA) is a reliable method for assessing VA status of the population. In this context, high performance liquid chromatography (HPLC) has become the method of choice for VA analysis. OBJECTIVES: We describe the VAD profile of Moroccan children and women, obtained by specific HPLC analysis of VA. SUBJECTS AND METHODS: The study involved 2457 participants: 1453 children and 1004 women from different areas of Morocco. Vitamin A was assessed by HPLC. Validation was conducted by comparison of the analysis of VA achieved in two laboratories. RESULTS: The prevalence of VAD in children (n = 1453) was 40.4% [37.8% - 43.0%] against 10.9 % [8.4% - 14.1%] of women in the Gharb (n = 494) and 6.3 % [4.4% - 8.8%] in women of Doukkala (n = 510). The VA distribution of the children was low compared to distributions among women. Children had significantly lower VA levels (0.773 +/- 0.252 micromol/l) than women of Gharb (1.17 +/- 0.43 micromol/l) and women of Doukkala (1.308 +/- 0.453 micromol/l). Analytical performance was good: the reproducibility and recovery rates were respectively 1.97 % and 98.7 +/- 3.1 %. The comparison of the mean VA and the cases of VAD obtained by the two laboratories showed no significant difference. CONCLUSION: Vitamin A deficiency is severe among children and moderate among women. The absence of the stratum having comfortable averages in vitamin A (plasma VA>3.49 micromol/l) indicates likely inadequate alimentary habits for a suitable vitamin A intake. The use of HPLC analysis of vitamin A, on a large scale for the first time in Morocco, has demonstrated that vitamin A deficiency is a public health problem. The recycling of mobile phase has reduced the costs of HPLC. This analytical method is useful in the evaluation of intervention programs against vitamin A deficiency and in future epidemiological studies. PMID- 15126187 TI - Maternal and fetal outcome of gestational diabetes mellitus in Mulago Hospital, Uganda. AB - OBJECTIVE: To determine the maternal and foetal outcomes in mothers with gestational diabetes mellitus attending antenatal clinics in Mulago Hospital Kampala Uganda. DESIGN: This was a cohort study. SETTING: Mulago Hospital antenatal clinics. PARTICIPANTS: Ninety mothers with gestational ages between 24 32 weeks were recruited from April to September 2001.They were followed up to the time of delivery. The WHO criterion for the diagnosis of gestational diabetes was used. Thirty mothers with a 2 hrs post prandial capillary blood sugar more than 140 mg/dl were the exposed group and 60 mothers with less than 140 mg/dl were the unexposed group. Blood sugar was measured using a one touch glucometer. OUTCOME VARIABLES: Socio demographic characteristics, maternal complications, mode of delivery and the foetal outcomes. RESULTS: The mean age of mothers in both groups was similar: 28.6 years vs 27.5 years. Both groups had similar body mass index more than 26. The mothers with gestational diabetes mellitus (GDM) were four times more likely to have hypertensive disease(p=0.04) and nine times more likely to have vaginal candidiasis(p=0.002). The modes of delivery were similar in both groups but genital injuries were more common among mothers with GDM. The indications of Caesarian section in mothers with GDM were two times more likely to be due to big babies and obstructed labour. The babies for mothers with GDM were more likely to be macrocosmic, still born, and have shoulder dystocia than those of normal mothers. CONCLUSION: Gestational diabetes mellitus exists in Uganda and is associated with adverse maternal and foetal outcomes. There is need to routinely screen mothers for gestational diabetes in this environment. PMID- 15126188 TI - Salivary gland tumors in Uganda: clinical pathological study. AB - BACKGROUND: The incidence of salivary gland tumors is claimed to be influenced by geographical and racial factors. The pathological classification and nomenclature of salivary gland tumors as defined by WHO classification (1991), is accepted world-wide but little is available in the literature regarding the spectrum of salivary gland tumors in Africa in the basis of this classification. Such efforts would allow comparison and justify any differences between the black African population and the rest of the world. OBJECTIVE: To outline the clinicopathological features of salivary gland tumors in Uganda. SETTING: Makerere University, Faculty of Medicine, Department of Pathology. METHODS: All epithelial tumors from major and minor salivary glands accessioned from 1979 to 1988 were analyzed in respect to sex and age of patients, anatomical location of the tumor and histological type. The histological diagnosis of each individual tumor was based on the 1991 WHO classification of salivary gland tumors. RESULTS: During the span of 10 years, 268 cases of salivary gland tumors were diagnosed. Of these, 113 (42.2%) were males, 148 (55.2%) females and in the remaining seven (2.6%) cases, the sex was not specified. The age range of the 247 patients with recorded ages was from 0.5 to 80 years. The mean age at diagnosis was 38.1 (SD =17.03) with the median of 38.0 years. Thirty four percent of tumors originated from the parotid, 33.2% from the submandibular and 32.8% from minor salivary glands. No tumor was implicated from the sublingual gland. There were a total of 125 (46.6%) malignant tumors and 143 (53.4%) benign tumors. The mean age of patients with malignant lesions (43.1 years; SD=16.75; median=44.00 years) was 9.6 years older than those with benign tumors (mean=33.5 years; SD=16.0; median=30.00 years). Pleomorphic adenoma was the most common benign tumor (74.8%), followed by myoepithelioma (9.8%). No Whartin's tumor was encountered. The malignant tumors were dominated by adenoid cystic carcinoma (28.8%) followed by mucoepidermoid carcinoma (21.6%). CONCLUSION: The pattern of distribution of salivary gland tumors in black African population seems to differ from that of Western series in that; i) females are more affected than males, ii) there is a low proportion of tumors from the parotid gland and high proportion of tumors from the submandibular and minor salivary glands, iii) the parotid and minor salivary gland tumors have more probability of being malignant than those tumors from the submandibular gland iv) the newly categorized pathological entities are common and v) Whartin's tumor is extremely rare in black African population. PMID- 15126189 TI - Temperament characteristics of street and non-street children in Eldoret, Kenya. AB - OBJECTIVE: To examine the interaction of temperament and environment and how these impact on the psychological function of street children and non-street children in Eldoret Kenya. METHOD: This was a cross-sectional study conducted in Eldoret town. A Revised Dimensions of Temperament Survey (DOTS-R) a 54 item, factor-analytically-developed self-report instrument that measures nine temperament dimensions was used. RESULTS: The statistical analysis showed that the largest significant level was on the approach/withdrawal dimension (F = 12.38, p<.001) the activity level-sleep (F = 4.20, p<.01) and the task orientation (F = 3.62, p<.01) dimensions were next in rank in terms of significance. The highest mean score on activity level-general (17.88) and the activity level-sleep (9.65) were in the "of" the street children that is consistent with their scavenging lifestyle and sleeping patterns. They have to be vigilant when sleeping on shop verandas, run down buildings, and trash-bins. CONCLUSION: These results support earlier research on street children. Counter to public opinion and hostility, the children are resilient, adaptable and flexibile in the face of adversity and remaining well adjusted as individuals. PMID- 15126191 TI - Gender issues in contraceptive use among educated women in Edo state, Nigeria. AB - BACKGROUND: While traditional contraception is widely used, in southern Nigerian modern contraception is a relatively recent phenomenon. Modern contraception is more wide spread among the educated and sexually active youth in Nigeria. Few studies have been done on contraception among educated women in Nigeria. OBJECTIVES: This study was carried out in December 2000 to determine factors that influence the choice of contraceptives among female undergraduates at the University of Benin and Edo State University Ekpoma. METHODS: Data was collected from a sample of 800 female undergraduates matched ethnic group, socio economic status, religion and rural urban residence. Subjects were selected by proportional representation and the instrument used was closed ended questionnaire. RESULTS: The responses obtained were analysed using Spearman Rank Correlation co-efficient and regression analysis. Findings revealed the highest correlation for availability (r =.96) vis a vis the use of various types of artificial contraceptive and cost (r =.96), next was safety (r =.95) and effectiveness (r =.95). Others were peer group influence (r =.80) and convenience (r =.77). CONCLUSION: An important step in improving women's reproductive health is the involvement of men. Health programmes should conduct campaigns to educate men about reproductive health and the role they can assume in family planning. PMID- 15126190 TI - Health providers' counselling of caregivers in the Integrated Management of Childhood Illness (IMCI) programme in Uganda. AB - BACKGROUND: IMCI was launched in Uganda in June 1995 and has so far been implemented in most districts. However, reports indicate that counselling is poorly performed and that health providers find IMCI counselling the most difficult component to implement. OBJECTIVES: The study was carried out to assess IMCI-trained health providers' counselling of caregivers and to determine factors that facilitate or constrain counselling. METHODS: A cross-sectional study utilizing quantitative and qualitative methods was carried out in 2000 in 19 health units in Mukono District. The study involved 37 health providers in 161 IMCI counselling sessions. RESULTS: Health providers performed well in assessing the child's problem (85%); listening (100%); use of simple language (95%); use of kind tone of voice (99%); showing interest in caregivers (99%); giving feeding advice (76%); and giving advice on return immediately (78%), for follow up (75%), and for immunization (97%). Performance was poor in praising the caregivers (43%); asking feeding questions (65%); explaining feeding problems (50%); explaining health problems (62%); advising on fluid intake (44%); advising on medication (61%), and using mothers' cards (44%). Most health providers (99%) did not address caregivers' health problems. Cadre of health provider, IMCI experience, number of supervisory visits and praise of health provider were independent predictors of using mothers' cards, advising on medication, inviting questions from caregivers, and advising on fluid intake respectively. Twelve percent of the children were referred but most health facilities did not have drugs to treat the children before referring them. CONCLUSIONS: The performance of health providers was good in 9 out of 20 IMCI counselling items, and cadre of health provider, IMCI experience, number of supervisory visits and praise of health provider were associated with IMCI counselling. Improvements in IMCI counselling could be achieved through emphasis on use of IMCI job aids; strengthening support supervision and providing positive feedback to health providers. The issue of availability of pre-referral drugs should be addressed by ensuring that these drugs are part of the essential drug kit. Finally, health providers should be trained and encouraged to address the health of the caregivers as well. PMID- 15126192 TI - Manifestations, quality of emergency care and outcome of severe malaria in Mulago Hospital, Uganda. AB - BACKGROUND: About 100,000 children die annually from severe malaria in Uganda and more than 75% of health unit based deaths occur within 24 hours of admission. Most of these deaths are associated with poor resuscitation systems, delays within the units by health workers and lack of essential drugs and supplies. OBJECTIVE: To describe the manifestations and quality of care children with severe malaria receive in Mulago Hospital Paediatric emergency unit and evaluate its impact on outcome. METHODS: A cohort of 784 children with severe malaria was recruited at admission and followed up. Selected measures of quality were the exposure factor and death, the outcome measure. RESULTS: Only 22.5% of the children were brought at night. The commonest defining manifestations were severe anaemia (39.4%), respiratory distress (17.1%), multiple generalized convulsions (13.3%), hypoglycaemia (11.4%) and cerebral malaria (7.2%). Over 50% lacked an essential drug or supply needed for resuscitation and 23.4% were seen within 1 hour of arrival. Commonly lacking items were intravenous cannulae (53.1%) syringes (23.3%) and blood transfusion sets (15.0%). Children brought at night took a shorter time before being seen by a doctor (1.9 SD 2.4 vs 2.5 SD 2.0 hours, p=0.002), received the first dose of quinine earlier (4.1 SD 3.2 vs 5.2 SD 3.2 hours, p<0.0001), fewer lacked essential drugs and supplies (45% vs 57.9%, p=0.003) and fewer died (0.6% vs 3.8%, p=0.028). Children who lacked an item for resuscitation took 30 minutes longer to receive the first dose of quinine. Caretaker satisfaction was predictive of mortality in the unit. CONCLUSIONS: Quality of care for severe malaria in Mulago paediatric emergency unit is still poor although nighttime services are comparatively better. Caretakers buy at least one resuscitation item in over 50% of cases and their level of satisfaction is predictive of mortality. RECOMMENDATIONS: The unit should set targets for quality improvement to include increased staffing and supplies, a time limit within which children should be seen and measures of decongestion. Determination of blood sugar in patients with severe malaria should be made a basic requirement. PMID- 15126193 TI - Treatment of fissure in ano- revisited. AB - INTRODUCTION: Fissure in ano is a troubling and painful condition that affects a great majority of the population world over. The nature and anatomy of fissure in ano is quite clear, and much is known about the various predisposing and contributing factors that lead to initiation and progression of the disease. The preferred method of treating them, one that results in optimal clinical results and the least pain and inconvenience to the patient, however, has been open to debate. METHODS: This paper outlines a brief account of the present scenario of different techniques available for the treatment of chronic anal fissure. CONCLUSION: Medical manipulation of the internal sphincter should be a first-line treatment in anal fissure. When this fails or fissures recur, lateral subcutaneous internal sphincterotomy should be the preferred options for the treatment of chronic fissure in ano. Nevertheless, all the option should be presented to the patient with complete information about the method, cure rates, complications, and recurrence of the disease. PMID- 15126194 TI - Moyer's method of mixed dentition analysis: a meta-analysis. AB - BACKGROUND: Mixed dentition analysis forms an essential part of an orthodontic assessment. Moyer's method which is commonly used for this analysis is based on data derived from a Caucasian population. The applicability of tables derived from the data Moyer used to other ethnic groups has been doubted. However no meta analyses have been done to statistically prove this. OBJECTIVE: To assess the applicability of Moyer's method in different ethnic groups. STUDY DESIGN: A meta analysis of studies done on other populations using Moyer's method. METHOD: The seven articles included in this study were identified by a literature search of Medline (1966-June 2003) using predetermined key words, inclusion and exclusion criteria. 195 articles were reviewed and meta-analyzed. RESULTS: Overall the correlation coefficients were found to be borderline in variation with a p-value of 0.05. Separation of the articles into Caucasian and Asian groups also gave borderline p-values of 0.05. CONCLUSION: Variation in the correlation coefficients of different populations using Moyer's method may fall either side. This implies that Moyer's method of prediction may have population variations. For one to be sure of the accuracy while using Moyer's method it may be safer to develop prediction tables for specific populations. Thus Moyer's method cannot universally be applied without question. PMID- 15126195 TI - Gender inequality and domestic violence: implications for human immunodeficiency virus (HIV) prevention. AB - Domestic violence and human immunodeficiency virus (HIV) infection are problems of great public health worldwide, especially sub-Saharan Africa and much of the developing countries. This is due to their far reaching social, economic and public health consequences. The two problems have gender inequality and gender power imbalances as the driving force behind the "epidemics". HIV infection is mainly acquired through heterosexual relations, which themselves are greatly influenced by socio-cultural factors, underlying which are gender power imbalances. Unfortunately gender relations, and gender issues in general, have not been given much emphasis in the medical perspective, especially in efforts for prevention and control of HIV infection. There is thus a need to mainstream gender relations in reproductive health. This article aims at emphasizing the intersection between domestic violence, gender inequality and HIV infection. PMID- 15126196 TI - Francis Omaswa: From cardiothoracic surgeon to village health advocate. Interview by Charles Wendo. AB - Uganda's Director General of Health Services, Prof. Francis Omaswa says that abolition of user fees in his country has enabled more people to access health services. "Development partners now agree that it was a mistake to require people to pay for health services. the rest of the world should go that way" From high tech practice in the UK and Nairobi, Prof Omaswa established a model rural operation theatre in a poor rural area in Ngora, Eastern Uganda that became the envy of East Africa. He established model village health teams to provide the backbone of a district health system. When he became the head of Uganda's ministry of health he pushed for these teams country wide. This, together with President Museveni's open and multi sectoral approach to the HIV/AIDS epidemic, has contributed to Uganda's success story: HIV prevalence rates have plummeted from 30% to 5%. Charles Wendo caught up with him and gives a rare glimpse into this remarkable son of Africa: surgeon, public health advocate with rare leadership skills. PMID- 15126199 TI - Dental informatics: a work in progress. AB - Dental informatics is a young scientific discipline that is undergoing continual maturation. Its literature is estimated to consist of approximately 600 papers published between 1975 and 2003, and it is currently growing at a rate of about 50 papers annually. While interest in the discipline is growing, the number of core contributors to dental informatics research remains relatively small. Two major questions for the discipline are: What are the research challenges that dental informatics faces today? and How can the discipline be strengthened and positioned to maximize its success in addressing those challenges? Progress toward research challenges formulated more than ten years ago has been varied. While many new technologies have become available for clinical dental practice, research, and education, many fundamental problems remain to be addressed with informatics research. Recommendations to augment the research capacity in dental informatics include creating a stronger worldwide dental informatics research community, drawing more biomedical informatics researchers to dental research areas, providing career opportunities for dental informatics researchers, addressing grand challenges together as a community, and recruiting subsequent generations of dental informaticians. PMID- 15126200 TI - Grand challenges in dental informatics. AB - When truly significant scientific challenges are overcome, it profoundly changes the daily activities, as well as the future research activities, of everyone involved in the related field. By identifying and describing the grand challenges facing a scientific field, we can help funding agencies identify and prioritize projects for support, stimulate and encourage new investigators to work on these intellectual and technological challenges, and help define the field itself. In this article, we present an informatics-oriented, future-patient-care scenario, then describe a series of applications and the related informatics grand challenges facing the dental field today. New techniques and technologies to help us overcome these challenges would facilitate the development of truly monumental applications, such as a comprehensive electronic oral health record, an automated dental treatment planning system for all diagnoses, or a system to profile patient risk for chronic oral diseases. PMID- 15126201 TI - A preliminary analysis of the dental informatics literature. AB - Dental informatics is an emerging discipline applying computer and information science to dental practice, research, education, and management. To date, the dental informatics research literature has not been comprehensively reviewed. This study reports an initial analysis of the dental informatics literature. We developed an initial, comprehensive retrieval strategy to locate dental informatics citations in MEDLINE (1966-April 2003), including three concepts: dentistry, computers, and research. After refinement of the search, we manually classified the final set into four categories: (1) non-dental; (2) dental, but neither dental informatics nor IT-related; (3) dental informatics; and (4) IT in dentistry. We analyzed informatics and IT-related citations regarding their distribution across journals, growth rate, the number of authors and their publication frequency, and content as expressed by Medical Subject Headings (MeSH). The final set of citations (n = 3872) consisted of: 12% non-dental articles; 59% dental, but not informatics- or IT-related articles; 16% informatics-related articles; and 13% IT-related articles. Informatics-related citations appeared in 176 journals, and IT-related citations in 206 journals. Approximately 50 papers are currently published in both categories yearly. While a great many authors have contributed to this literature, very few have published more than three papers. Main topics of articles included "Imaging and Image Processing", "Computer-aided Diagnosis and Therapy", "Computer-aided Instruction", and "Other". The dental informatics literature is small, but growing. Imaging and image processing predominate as research topics. PMID- 15126202 TI - Defining biomedical informatics competency: the foundations of a profession. AB - Is biomedical informatics a science or a profession? This question has been asked of many members in the biomedical informatics community, yet we still lack a response that galvanizes our community. We debate the issues over lunch. We create long, multi-threaded e-mail discussions, we write papers on the topic, and still we aren't able to convince ourselves-let alone the rest of the scientific community. In this paper, I will describe a curriculum model for biomedical informatics and research that is developing at Columbia University, Department of Biomedical Informatics (DBMI). We believe that a strong educational foundation creates competent professionals who, in turn, comprise a bioinformatics culture. The outcome of DBMI's curriculum design and competency project will be a set of biomedical informatics competencies which we believe will define the core knowledge and skills of the field. PMID- 15126203 TI - Biomedical informatics training for dental researchers. AB - Dental researchers collaborating closely with biomedical informaticians have achieved many advances in oral health research, such as in mapping human genetics and addressing oral health disparities. Advances will continue to increase as dental researchers and biomedical informaticians study each others' disciplines to increase the effectiveness of their collaborative research. The combined skills will greatly increase the effectiveness of dental research. This manuscript summarizes the core of biomedical informatics curriculum (biomedical informatics knowledge, data management, and software engineering) for dental research. It also summarizes the obstacles that must be overcome for all dental research students to receive the training in biomedical informatics they require. These issues are: a lack of biomedical informatics faculty, a lack of biomedical informatics courses, and a lack of accreditation standards. Last, intra- and inter-institutional collaboration solutions are described. PMID- 15126204 TI - Issues and strategies for faculty development in technology and biomedical informatics. AB - Biomedical informatics and technology are becoming important components of dental education. The tools and techniques now available have the potential for significant impact on teaching and research by improving the way information is acquired, stored, retrieved, and managed. However, a gap exists between those who create, introduce, or implement the technology applications and the faculty in dental schools faced with the challenge of using it. For technology and informatics to thrive in the areas of didactic teaching, clinical teaching, and clinical practice, more than a select few must understand the potential applications. This paper provides an overview of the issues and strategies involved with faculty development for the use of technology in the educational setting. The discussion covers important reasons for developing faculty competence in technology applications, significant barriers to faculty development in this area, and several strategies designed to overcome these barriers. PMID- 15126205 TI - Adaptive hypermedia: a new paradigm for educational software. AB - Traditional online dental education courses follow the broadcast paradigm which centers on the teacher, not the student. This one-size-fits-all approach resembles a mass-production idea which cannot take individual learner characteristics into account. Most online course designs do not address the issue that users with different goals and knowledge may be interested in different pieces of information about a topic. Adaptive hypermedia (AH) is an emerging field in education research which investigates how computer systems can overcome this problem. AH can be applied to any course content. This learner-centered approach first considers the learning goal(s), then evaluates the user's abilities and determines the individual learning style, to structure and tailor the curriculum most efficiently. The presented AH environment exploits various concepts of AH. The system collects data to create a model of the individual user, which is continuously refined based on test results throughout the course. The system then adapts the learning material dynamically, using active and passive curriculum sequencing and adaptive presentation. PMID- 15126206 TI - Informatics systems to assess and apply clinical research on dental restorative materials. AB - Dental biomaterials are used clinically for one or more of the following purposes: to restore function, to enhance esthetics, and to prevent or arrest demineralization of tooth structure. Studies of the clinical performance of restorations and prostheses made from these materials have generally focused on quality assessment and survival statistics. Data from these studies should provide probabilities of specific treatment outcomes that are useful for practicing dentists. However, the utility of these data is limited by the lack of national and international standards for assessing these clinical outcomes. Standardized approaches toward clinical informatics and treatment-decision analysis are urgently needed to minimize the variability of clinical outcomes reported in publications associated with direct and indirect restorative materials used for dental restorations and prostheses. PMID- 15126207 TI - Informatics challenges in tissue engineering and biomaterials. AB - Both tissue engineering and biomaterials have made tremendous strides recently, yet major questions remain unanswered. Tissue-engineered products have come to the market; others are in development. A fundamental issue that informatics could address for tissue engineering is to describe and to predict the cascade of biochemical and cellular reactions that occur as a function of time and implant material: surface texture, microporosity; pore size, density, and connectivity; and three-dimensional configuration. Behavior of ceramics, a subset of tissue engineering scaffold materials and a mainstay of dental restorations, has been studied extensively for very thin layers and for thicknesses greater than 2 mm. Until recently, little has been known about dentally relevant thickness of 1-2 mm. Results have been surprising and are continuing to develop. Still, at least one fundamental question remains that could be addressed by informatics techniques: Where, along the spectrum of flat-polished material to 10-year clinical in vivo study, can we test to predict clinical performance of all ceramic crowns accurately? PMID- 15126208 TI - Spectrophotometric analysis of all-ceramic materials and their interaction with luting agents and different backgrounds. AB - In this study, two All-Ceramic (AC) materials--Empress 2 (EMP) (Ivoclar Vivadent AG, Schaan, Liechtenstein) and In-Ceram ALUMINA (ICA) (Vita Zahnfabrik, Bad Sackingen, Germany)--were analyzed, along with the effects of 3 luting agents viz. Zinc Phosphate cement (ZNPO, PhospaCEM PL, Ivoclar Vivadent AG, Schaan, Liechtenstein), Glass Ionomer Cement (GIC, Ketac-Cem Radiopaque, ESPE Dental AG, Seefeld, Germany), and Compolute (COMP, ESPE Dental AG, Seefeld, Germany)--on the final color, using the CIELab system. Color differences (DeltaL, Deltaa, Deltab, and DeltaE) were calculated for samples with luting agents and for samples without luting agents with standard white and black backgrounds, with the use of a spectrophotometer, Luci 100 (Dr. Lange, Berlin, Germany). One-way ANOVA for DeltaL, Deltaa, Deltab, and DeltaE within both the AC systems, with and without luting agents, showed significant contributions of the background (p < 0.05). EMP was seen to be more translucent than ICA. Darker ceramics showed less color variation. Luting agents altered the final color of the restoration. ZNPO was least translucent, followed by GIC and COMP. Marginal increases in thicknesses of ICA samples (0.4 mm) do not show a statistically significant color difference. No method exists to predict the outcome of an AC restoration based on consideration of the luting agent and the background color. PMID- 15126209 TI - Computational models of oral and craniofacial development, growth, and repair. AB - This paper illustrates how biological and clinical problems stimulate research in biomedical informatics and how such research contributes to their solution. The computational models described use techniques from Logic Programming, Machine Learning, Computer Vision, and Biomathematics. They address problems in the development, growth, and repair of oral and craniofacial tissues arising in cell biology, clinical genetics, and dentistry. At the micro-level, the dynamic interaction of cells in the oral epithelium is modeled. At the macro-level, models are constructed of either the craniofacial shape of an individual or the craniofacial shape differences within and between healthy and congenitally abnormal populations. In between, in terms of scale, there are models of normal dentition and the use of computerized expert knowledge to guide the design of dental prostheses used to restore function in partially edentulous patients. PMID- 15126210 TI - Designing clinically useful systems: examples from medicine and dentistry. AB - Despite promising results in medical informatics research and the development of a large number of different systems, few systems get beyond a prototype state and are really used in practice. Among other factors, the lack of explicit user focus is one main reason. The research projects presented in this paper follow a user centered system development approach based on extensive work analyses in interdisciplinary working groups, taking into account human cognitive performance. Different medical and health-care specialists, together with researchers in human-computer interaction and medical informatics, specify future clinical work scenarios. Special focus is put on analysis and design of the information and communication flow and on exploration of intuitive visualization and interaction techniques for clinical information. Adequate choice of the technical access device is made depending on the user's work situation. It is the purpose of this paper to apply this method in two different research projects and thereby to show its potential for designing clinically useful systems that do support and not hamper clinical work. These research projects cover IT support for chairside work in dentistry (http://www.dis.uu.se/mdi/research/projects/orquest) and ICT support for home health care of elderly citizens (http://www.medsci.uu.se/mie/project/closecare). PMID- 15126211 TI - Decision support at the point of care: challenges in knowledge representation, management, and patient-specific access. AB - Many applications in a clinical information system can benefit from the incorporation of medical knowledge to provide patient-specific, point-of-care decision support. These include computer-based provider order entry, referral, clinical result interpretation, consultation, adverse event monitoring, scheduling, shared patient-doctor decision-making, and generation of alerts and reminders, among others. To be executable, knowledge must be represented in the form of rules, constraints, calculations, guidelines, and other logical/algorithmic formats. The main difficulty is that the integration of such knowledge into clinical applications, when it occurs, tends to be very system- and application-specific, often encoded in a programming language, or even in the formating specifications of a user interaction display. Also, the data references and services invoked are highly dependent on the system/platform and electronic medical record implementation. This makes it difficult and time-consuming to encode authoritative evidence-based knowledge, severely limits the ability to disseminate and share successes, and hampers efforts to review and update the logic as medical knowledge changes. Solutions to this problem involve the development of standards-based representations for medical knowledge, and tools for authoring/editing, dissemination, adaptation to local environments, and execution. Numerous approaches are being pursued, all of which will be described in this presentation. PMID- 15126212 TI - AIDA: web agents in dental treatment planning. AB - The objective of the AIDA project (Artificial Intelligent Dental Agents, http://aida.uni-hd.de) is the analysis of dental decision-making, the design of a computer-based decision support system, as well as the testing of the decision structure in interactions with dental experts, practicing dentists, and patients. The planning of the solution alternatives for an individual patient is based on a top-down structure for dental decision-making, aiming at a standardization of the argumentation. From a theoretical point of view, decision support can be provided only for anticipated decisions (planning). Moreover, only parts of these anticipated decisions can be supported. Accordingly, a separation of these partial aspects has to take place before one is able to build decision support systems. For prosthetic dentistry, clinicians have been shown how to use individual patient findings to sketch the possible treatment alternatives and later derive guidelines for the treatment. The planning module for fixed prostheses has already been integrated into a software agent. Planning modules for other types of prostheses are currently specified, implemented, and verified. PMID- 15126213 TI - The role of information technology and informatics research in the dentist patient relationship. AB - A high-value doctor-patient relationship is based on a set of parameters which include the interpersonal relationship between the patient and the doctor. Based on the Primary Care Assessment Survey model, measures of the interpersonal relationship are associated with communication, interpersonal care, contextual knowledge of the patient, and trust. Despite the proven value of the doctor patient relationship, current trends indicate that the quality of these relationships is on the decline. The advent of communication and information technologies has greatly affected the way in which health care is delivered and the relationship between doctors and patients. The convergence of communication and information technology with biomedical informatics offers an opportunity to affect the character of the doctor-patient relationship positively. This paper examines the intersection of the key features of the doctor-patient relationship and a variety of Internet-based, clinical, and administrative applications used in dental practice. This paper discusses the role of dental informatics research vis-a-vis the doctor-patient relationship and explores how it may inform the next generation of information technologies used in dental practice. PMID- 15126214 TI - Health services research. AB - The major barriers to the collection of primary population-based dental services data are: (1) Dentists do not use standard record systems; (2) few dentists use electronic records; and (3) it is costly to abstract paper dental records. The value of secondary data from paid insurance claims is limited, because dentists code only services delivered and not diagnoses, and it is difficult to obtain and merge claims from multiple insurance carriers. In a national demonstration project on the impact of community-based dental education programs on the care provided to underserved populations, we have developed a simplified dental visit encounter system. Senior students and residents from 15 dental schools (approximately 200 to 300 community delivery sites) will use computers or scannable paper forms to collect basic patient demographic and service data on several hundred thousand patient visits. Within the next 10 years, more dentists will use electronic records. To be of value to researchers, these data need to be collected according to a standardized record format and to be available regionally from public or private insurers. PMID- 15126215 TI - Using information technology and community-based research to improve the dental health-care system. AB - It is commonly acknowledged that the United States' health-care system produces some of the finest care in the world for some people but fails to meet the needs of others. The Institute of Medicine (IOM) issued six aims for a redesigned health-care system, that it be: safe, effective, patient-centered, timely, efficient, and equitable. The purpose of this paper is to use an ongoing community-based study to illustrate current problems in the provision of oral health services that could be addressed through information technology. Appropriate use of information technology can assist dental schools and clinics in community-based clinical outcomes research needed to assemble the evidence base for improving oral health care. This conference serves as an important steppingstone to establish a means for information technology to improve the community's oral health. PMID- 15126216 TI - Overview of bioinformatics and its application to oral genomics. AB - The "informatics revolution" in both bioinformatics and dental informatics will eventually change the way we practice dentistry. This convergence will play a pivotal role in creating a bridge of opportunity by integrating scientific and clinical specialties to promote the advances in treatment, risk assessment, diagnosis, therapeutics, and oral health-care outcome. Bioinformatics has been an emerging field in the biomedical research community and has been gaining momentum in dental medicine. This area has created a steady stream of large and complex genomic data, which has transformed the way a clinical or basic science researcher approaches genomic research. This application to dental medicine, termed "oral genomics", can aid in the molecular understanding of the genes and proteins, their interactions, pathways, and networks that are responsible for the development and progression of oral diseases and disorders. As the result of the Human Genome Project, new advances have prompted high-throughput technologies, such as DNA microarrays, which have become accepted tools in the biomedical research community. This manuscript reviews the two most commonly used microarray technologies, basic microarray data analysis, and the results from several ongoing oral cancer genomic studies. PMID- 15126217 TI - Genetic and molecular characterization of a dental pathogen using genome-wide approaches. AB - Actinobacillus actinomycetemcomitans causes periodontitis, a costly chronic infection that affects a large number of patients. The pathogenesis of this dental infection is a multifactorial process that results in a serious degenerative disease of the periodontium. Although significant progress has been achieved after the identification of this Gram-negative bacterium as the etiological agent of this infection, much remains to be done to understand in detail the bacterial factors and host-pathogen interactions involved in the pathogenesis of this disease. Classic research approaches have resulted in the identification of important virulence factors and cellular processes, although they have provided a rather narrow picture of some of the steps of this complex process. In contrast, a much wider picture could be obtained with the application of tools such as bioinformatics and genomics. These tools will provide global information regarding the differential expression of genes encoding factors and processes that lead to the pathogenesis of this disease. Furthermore, comparative genomics has the potential of helping us to understand the emergence and evolution of this human pathogen. This genome-wide approach should provide a more complete picture of the pathogenesis process of this disease, and will facilitate the development of efficient diagnostic, preventive, and therapeutic measures for this disease. PMID- 15126218 TI - Comparative genomics and structure prediction of dental matrix proteins. AB - Non-collagenous matrix proteins secreted by the ameloblasts (amelogenin) and odontoblasts (osteocalcin) play important roles in the mineralization of enamel and dentin. In this study, comparative genomics approaches were used to identify the functional domains and model the three-dimensional structure of amelogenin and osteocalcin, respectively. Multiple sequence analysis of amelogenin in different species showed a high degree of sequence conservation at the nucleotide and protein levels. At the protein level, motifs (a sequence pattern that occurs repeatedly in a group of related proteins or genes), conserved domains, secondary structural characteristics, and functional sites of amelogenin from lower phyla were similar to those of the higher-level mammals, reflecting the high degree of sequence conservation during vertebrate evolution. Osteocalcin, produced by both odontoblasts and osetoblasts, also showed sequence similarity between species. Three-dimensional structure predictions developed by modeling of conserved domains of osteocalcin supported a role for glutamic acid residues in the calcium mineralization process. PMID- 15126219 TI - Bayesian machine learning and its potential applications to the genomic study of oral oncology. AB - With the completion of the Human Genome Project and the growing computational challenges presented by the large amount of genomic data available today, machine learning is becoming an integral part of biomedical research and plays a major role in the emerging fields of bioinformatics and computational biology. This situation offers unparalleled opportunities and unprecedented challenges to machine learning research in general and to Bayesian learning methods in particular. This paper outlines some of the opportunities and the challenges of this endeavor, it describes where the efforts of "cracking the code of life" can most benefit from a Bayesian approach, and it identifies some potential applications of Bayesian machine learning methods to the genomic analysis of squamous cell carcinomas of the head and neck. PMID- 15126220 TI - Dental data mining: potential pitfalls and practical issues. AB - Knowledge Discovery and Data Mining (KDD) have become popular buzzwords. But what exactly is data mining? What are its strengths and limitations? Classic regression, artificial neural network (ANN), and classification and regression tree (CART) models are common KDD tools. Some recent reports (e.g., Kattan et al., 1998) show that ANN and CART models can perform better than classic regression models: CART models excel at covariate interactions, while ANN models excel at nonlinear covariates. Model prediction performance is examined with the use of validation procedures and evaluating concordance, sensitivity, specificity, and likelihood ratio. To aid interpretation, various plots of predicted probabilities are utilized, such as lift charts, receiver operating characteristic curves, and cumulative captured-response plots. A dental caries study is used as an illustrative example. This paper compares the performance of logistic regression with KDD methods of CART and ANN in analyzing data from the Rochester caries study. With careful analysis, such as validation with sufficient sample size and the use of proper competitors, problems of naive KDD analyses (Schwarzer et al., 2000) can be carefully avoided. PMID- 15126221 TI - Retrieval and classification of dental research articles. AB - Successful retrieval of a corpus of literature on a broad topic can be difficult. This study demonstrates a method to retrieve the dental and craniofacial research literature. We explored MeSH manually for dental or craniofacial indexing terms. MEDLINE was searched using these terms, and a random sample of references was extracted from the resulting set. Sixteen dental research experts categorized these articles, reading only the title and abstract, as either: (1) dental research, (2) dental non-research, (3) non-dental, or (4) not sure. Identify Patient Sets (IPS), a probabilistic text classifier, created models, based on the presence or absence of words or UMLS phrases, that distinguished dental research articles from all others. These models were applied to a test set with different inputs for each article: (1) title and abstract only, (2) MeSH terms only, or (3) both. By title and abstract only, IPS correctly classified 64% of all dental research articles present in the test set. The percentage of correctly classified dental research articles in this retrieved set was 71%. MeSH term inclusion decreased performance. Computer programs that use text input to categorize articles may aid in retrieval of a broad corpus of literature better than indexing terms or key words alone. PMID- 15126223 TI - Caregiving in geriatric psychiatry. PMID- 15126224 TI - Family caregiving of persons with dementia: prevalence, health effects, and support strategies. AB - The authors summarize the dementia caregiving literature and provide recommendations regarding practice guidelines for health professionals working with caregivers. Family caregiving of older persons with disability has become commonplace in the United States because of increases in life expectancy and the aging of the population, with resulting higher prevalence of chronic diseases and associated disabilities, increased constraints in healthcare reimbursement, and advances in medical technology. As a result, family members are increasingly being asked to perform complex tasks similar to those carried out by paid health or social service providers, often at great cost to their own well-being and great benefit to their relatives and society as a whole. The public health significance of caregiving has spawned an extensive literature in this area, much of it focused on dementia caregiving because of the unique and extreme challenges associated with caring for someone with cognitive impairment. This article summarizes the literature on dementia caregiving, identifies key issues and major findings regarding the definition and prevalence of caregiving, describes the psychiatric and physical health effects of caregiving, and reviews various intervention approaches to improving caregiver burden, depression, and quality of life. Authors review practice guidelines and recommendations for healthcare providers in light of the empirical literature on family caregiving. PMID- 15126225 TI - Use of cognitive enhancement medication in persons with Alzheimer disease who have a family caregiver: results from the Resources for Enhancing Alzheimer's Caregiver Health (REACH) project. AB - OBJECTIVE: Aging populations show increased prevalence of cognitive impairment and dementia. Recent efficacy studies report on prescription medications and herbal preparations that affect cognitive functioning, but the prevalence and correlates of cognitive-enhancement (CE) medication use among community-dwelling older persons is not well studied. The authors examined the frequency and appropriateness of use, the importance of a family caregiver in medication decisions for dementia patients, and differences in access to medical care. METHODS: REACH is a multisite feasibility study of several approaches to reducing the negative impacts of caregiving on those living with a family member with dementia. Data on medication use by care-recipients were collected at baseline and 1 year later. RESULTS: At baseline, 31% of 1,222 care-recipients were using a CE medication. Factors independently related to CE use were age, education, functional status, and caregiver vigilance. Within 1 year, 14% started and 30% quit taking CE. Care-recipients more likely to be Starters had spouse-caregivers, more education, and fewer baseline ADL impairments. Quitters had more ADL deficits at baseline and became less able to perform ADL at follow-up than those who continued on CE. CONCLUSIONS: CE medication use among dementia patients with a family caregiver is relatively common, though there is substantial geographic variability. Our findings are mixed with respect to appropriate use of CE medications, suggesting areas for physician education. Our data indicate the importance of the caregiver in CE medication use and suggest that there may be disparities in access to healthcare among people with cognitive impairment. PMID- 15126226 TI - Influence of age of patients who wish to die on treatment decisions by physicians and nurses. AB - OBJECTIVE: There is an ongoing debate on whether the wish to die in old age is a normal phenomenon and should be accepted as opposed to similar wishes in younger ages. The authors asked to what extent the age of a patient influences treatment decisions of physicians and nurses. METHODS: Real-life case vignettes of three persons ages 70 and over, with subthreshold depression and a wish to die, were presented to 19 physicians and 83 nurses, with the question of whether they would initiate psychopharmacological treatment or psychotherapy. For half of the participants, the real age of these persons was reduced by 20 years. Neither the participants nor the facilitators were aware of the focus of the study. RESULTS: The patient's age had a major influence on treatment decisions by physicians and nurses. The question of whether patients who expressed the wish to die should receive treatment, be it psychotherapy or intensive care, was answered with "yes" by both physicians and nurses significantly less frequently when the real age of the patient was known, whereas when a lower patient age (-20 years) was given, they more often saw the need for psychotherapy or other medical interventions. CONCLUSIONS: Age stereotypes have an influence on medical decision-making. Physicians and nurses must be trained in this respect in order to avoid maltreatment of elderly patients. PMID- 15126227 TI - Social support, depression, and functional disability in older adult primary-care patients. AB - OBJECTIVE: The authors asked whether social support and depression are independently associated with functional disability and examined the potential role of social support as a moderator in the depression-functional disability association. METHODS: Subjects were 305 patients age 60 years and over. Predictor variables were social support, depressive symptoms, and depression diagnosis. Dependent variables were the Instrumental Activities of Daily Living Scale, the Physical Self-Maintenance Scale, and the Physical Functioning subscale of the Medical Outcomes Study 36-Item Short-Form Health Survey. Authors used multiple regression analyses. RESULTS: Depressive symptoms and all dimensions of social support were independently associated with functional disability: the specifics of these relationships varied among types of social support and functional disability. Depression diagnosis was not independently associated with any functional disability measure. Social support (more instrumental help, more perceived satisfaction) moderated some depression diagnosis-functional disability associations, and one depressive symptom-functional disability association. CONCLUSIONS: The study hypotheses were partially confirmed. Different dimensions of social support have important and varied roles in the depression-functional disability dynamic. Future research is needed to further specify the complex relationships among depression, social support, and functional disability. PMID- 15126228 TI - What explains differences between dementia patients' and their caregivers' ratings of patients' quality of life? AB - OBJECTIVE: The authors assessed the magnitude of discrepancy between patients' and caregivers' ratings of the patients' quality of life and sought to determine whether the discrepancies are associated with patient characteristics, caregiver characteristics, or the type of relationship between the patient and caregiver. METHODS: A sample of 91 patients with mild-to-moderately severe dementia and their primary family caregiver rated five domains of the patients' subjective quality of life. RESULTS: Agreement between patients and caregivers was low. Caregivers rated patients' quality of life lower than patients rated their own in all five domains. Discrepancies between patients' and caregivers' ratings were not associated with the patients' cognitive performance, level of functioning, nor caregivers' reports of aggressive, attention-seeking, or sexually inappropriate behaviors, nor whether the caregiver lived with or was married to the patient. However, discrepancies were associated with level of caregiver burden and the patients' report of depressive symptoms. Patients with depression reported low quality of life, which matched caregivers' low rating of patients' quality of life. Caregivers who reported higher levels of burden rated patients' quality of life lower than did patients in all five domains of quality of life. CONCLUSIONS: Discrepancies between dementia patients' and their caregivers' ratings of the patients' quality of life are associated with increased levels of caregiver burden, rather than lower levels of patients' functioning. The results of this study support the direct assessment of mild-to-moderate dementia patients about their subjective quality of life. PMID- 15126229 TI - Vulnerable caregivers of Alzheimer disease patients have a deficit in beta 2 adrenergic receptor sensitivity and density. AB - OBJECTIVE: The chronic stress of caregiving may lead to sympathetic nervous system activation and immune suppression. beta(2)-adrenergic receptors are expressed on all immune cells and contribute to the stress-induced loss of immune cell function. The authors examined the effects of being a spousal caregiver of a patient with Alzheimer disease (AD) on the lymphocyte beta(2)-adrenergic receptor. METHODS: One hundred and six women and men, spousal caregivers and non caregivers, participated (mean age: 71.5 years). Caregivers were classified as either vulnerable or non-vulnerable on the basis of the amount of care required by the patient relative to the amount of respite the caregiver received during the previous 6 months. beta(2)-adrenergic receptor sensitivity (cyclic-AMP response to isoproterenol stimulation) and density (radioligand binding) were determined by use of whole lymphocytes. RESULTS: Vulnerable caregivers had reduced beta(2)-adrenergic receptor sensitivity and density when compared with their non-vulnerable counterparts or with non-caregivers. CONCLUSION: The findings indicate that for more vulnerable caregivers, the stress of caregiving leads to a loss of lymphocyte beta(2)-adrenergic receptors. This finding may be relevant to previous observations of clinically-relevant reduced immunity in highly stressed caregivers of AD patients. PMID- 15126230 TI - Recognition and management of depression in skilled-nursing and long-term care settings: evolving targets for quality improvement. AB - OBJECTIVE: Depression is a common disorder associated with suffering, morbidity, and mortality in nursing home residents. It is treatable, and improving the quality of treatment can have a major impact. METHODS: MPRO, Michigan's Quality Improvement Organization, initiated a quality-improvement project in 14 nursing facilities to improve the accuracy of assessments, targeting, and monitoring of care. Electronic Minimum Data Set (MDS) data and medical-record abstraction results were combined to form the analytic dataset. RESULTS: Findings from the baseline phase demonstrated that, according to medical and administrative records, 26% of newly admitted nursing home residents had symptoms of depression that were apparent at admission, and an additional 12% were recognized early in their stay. Eighty-one percent of residents with depression were receiving treatment on admission to the facility, and 79% of those with depression recognized by Day 14 were treated by then. CONCLUSIONS: These data demonstrate progress toward improving the initiation of treatment for depression in nursing homes; however, there are still opportunities for improving the quality of care and, especially, the quality of assessments. The authors recommend the addition of the Geriatric Depression Scale to the federally mandated MDS for cognitively intact patients. There could also be mechanisms to ensure that providers and facilities follow recommended practice guidelines. Initiating treatment with antidepressant medications should be followed with monitoring of residents to identify those who still have depressive symptoms and to modify or intensify their treatment. PMID- 15126231 TI - Some predictors of psychiatric consultation in nursing home residents. AB - OBJECTIVE: Despite the high rate of psychiatric disorders in nursing homes, research indicates that psychiatric consultation is requested infrequently. The authors sought to determine the rate of psychiatric consultation in a nursing home population and to assess what factors were related to a consultation request. METHODS: Subjects were recruited from a stratified random sample of 59 nursing homes across Maryland. All new admissions age 65 years and older from September 1992 through March 1995 were eligible for the study. A total of 2,285 subjects were included in the study. Variables examined were factor scores from the Cornell Scale for Depression in Dementia and the Psychogeriatric Dependency Rating Scale (Behavioral Subscale), nursing home characteristics, and whether the resident had a psychiatric consultation within 90 days of admission. RESULTS: Twenty percent of the residents (N=404) had a psychiatric consultation. There was no significant association with demographic variables. Behaviors that triggered a psychiatric consultation included agitation, physical/verbal abuse, wandering, and manic/destructive acts. A psychiatric consultation was also requested when residents displayed anxiety. Surprisingly, depression in retarded and psychotic residents did not trigger a psychiatric consult. CONCLUSION: As expected, behavioral problems and agitation are common reasons for a psychiatric consultation. However, the resident who is depressed, particularly the quiet or retarded depressed resident, may be overlooked. In this context, it is important for the nursing staff to recognize that lethargy and social withdrawal may be signs of depression, and a referral to a psychiatrist may be in order. PMID- 15126232 TI - Low incidence of delirium in very old patients after surgery for hip fractures. AB - OBJECTIVE: The authors sought to determine the incidence of delirium, its predisposing and precipitating factors, and its implications for rehabilitation outcomes and mortality in elderly patients. METHODS: This was a prospective cohort study, conducted in Gedera, Israel, from August 2001 to January 2002, with 137 consecutive patients over age 75, with hip fractures, who were admitted to the orthopedic section of the emergency department. They were evaluated at admission, 1 week after the surgery for hip fracture, and 1 month after surgery. The evaluation included assessments of delirium, cognitive, and functional status, and a wide range of demographic and clinical parameters. RESULTS: The cumulative incidence of delirium was 11.4%. No significant difference was found between delirium and non-delirium patients in terms of all demographic, socioeconomic, and perioperative parameters. Mild or moderate cognitive impairment before the fracture and four or more regular medications prescribed to the patient were the only predictive factors for the development of delirium in a multivariate model. Delirium was not a significant predictor of any rehabilitation outcome. CONCLUSION: Results documented that the incidence of delirium after hip fracture in elderly patients is much lower than was reported in several previous studies. Premorbid cognitive impairment was the most significant predisposing factor for the development of delirium. Thorough evaluation of earlier cognitive status could improve the probability of the diagnosis of delirium and pinpoint a limited group of patients for a delirium prevention approach. PMID- 15126233 TI - Cerebrovascular disease and late-life depression: a latent-variable analysis of depressive symptoms after stroke. AB - OBJECTIVE: The author used a structural-equation modeling approach to examine depression symptom endorsement between geriatric stroke patients and general medical geriatric patients. From the literature related to cerebrovascular disease and depression, he predicted that geriatric stroke patients would be more likely to endorse symptoms of withdrawal and affective flattening, and less agitation. METHODS: Data from 576 geriatric rehabilitation patients were utilized (N=176 with stroke). Confirmatory factor analysis of the Geriatric Depression Scale was conducted to determine the optimal factor structure for examining symptom endorsement differences between the two groups. MIMIC (Multiple Indicators, Multiple Causes) model analyses were incorporated to examine group differences in depression severity and symptom-cluster endorsement. RESULTS: Results indicate that stroke patients did not demonstrate greater depression severity. There were no differences in endorsement of depressive mood. However, stroke patients were more likely to endorse social withdrawal and less likely to endorse agitation symptoms. CONCLUSIONS: The current results suggest that geriatric stroke patients were more likely, after controlling for depression severity, to endorse greater social withdrawal and less agitation, but not depressed mood. These findings have implications for the types of symptoms evaluated in the context of geriatric stroke and depression. PMID- 15126234 TI - SSRIs do not cause affective blunting in healthy elderly volunteers. AB - OBJECTIVE: This study was undertaken to evaluate the effects of selective serotonin reuptake inhibitors (SSRIs) on affective experience in healthy older adults. METHODS: After 1 week of observation, normal elderly volunteers (age range: 65-84 years) were given placebo, paroxetine (10 mg-40 mg/day), or sertraline (50 mg-150 mg/day) for 3 weeks in a double-blind study. Paroxetine- and sertraline-treated subjects were analyzed together as the SSRI group (N=30). Volunteers were assessed weekly and recorded mood and events in a daily diary each evening. All data were analyzed with mixed-effects random-regression models. RESULTS: There were significant relationships between daily affect and events reported in the daily diary for the sample as a whole, with no differences between groups in mean slopes of positive or negative affect across the length of the study. There were no differences between groups in affective variability. However, the SSRI group, but not the placebo group, demonstrated a significant drug-dependent decrease in negative affect related to negative events. There were no other observed group differences on any other measure. CONCLUSION: Interpreting the results conservatively, they demonstrate that SSRIs are not associated with affective toxicity in elderly persons. PMID- 15126235 TI - Family caregiver functioning in late-life bipolar disorder. PMID- 15126236 TI - Impact of PPARgamma overexpression and activation on pancreatic islet gene expression profile analyzed with oligonucleotide microarrays. AB - Peroxisome proliferator-activated receptor-gamma (PPARgamma) serves as a target for the thiazolidinedione class of antidiabetic drugs and is an important regulator of adipose tissue differentiation. By contrast, the principal target genes for PPARgamma in the pancreatic islet and the impact of their induction on insulin secretion are largely undefined. Here, we show that mRNAs encoding both isoforms of rodent PPARgamma, gamma1 and gamma2, are expressed in primary rat islets and are upregulated by overexpresssion of the lipogenic transcription factor sterol response element-binding protein 1c. Unexpectedly, however, oligonucleotide microarray analysis demonstrates that graded activation of PPARgamma achieved with 1) the thiazolidinedione GW-347845, 2) transduction with adenoviral PPARgamma1, or 3) a combination of both treatments progressively enhances the expression of genes involved in fatty acid oxidation and transport. Moreover, maximal activation of PPARgamma1 reduces islet triglyceride levels and enhances the oxidation of exogenous palmitate while decreasing glucose oxidation, cellular ATP content, and glucose-, but not depolarization-stimulated, insulin secretion. We conclude that, in the context of the pancreatic islet, the principal response to PPARgamma expression and activation is the activation of genes involved in the disposal, rather than the synthesis, of fatty acids. Although fatty acid oxidation may have beneficial effects on beta-cell function in the longer term by countering beta-cell "lipotoxicity," the acute response to this metabolic shift is a marked inhibition of insulin secretion. PMID- 15126237 TI - C-peptide corrects endoneurial blood flow but not oxidative stress in type 1 BB/Wor rats. AB - Oxidative stress and neurovascular dysfunction have emerged as contributing factors to the development of experimental diabetic neuropathy (EDN) in streptozotocin-diabetic rodents. Additionally, depletion of C-peptide has been implicated in the pathogenesis of EDN, but the mechanisms of these effects have not been fully characterized. The aims of this study were therefore to explore the effects of diabetes on neurovascular dysfunction and indexes of nerve oxidative stress in type 1 bio-breeding Worcester (BB/Wor) rats and type 2 BB Zucker-derived (ZDR)/Wor rats and to determine the effects of C-peptide replacement in the former. Motor and sensory nerve conduction velocities (NCVs), hindlimb thermal thresholds, endoneurial blood flow, and indicators of oxidative stress were evaluated in nondiabetic control rats, BB/Wor rats, BB/Wor rats with rat II C-peptide replacement (75 nmol C-peptide.kg body wt(-1).day(-1)) for 2 mo, and diabetes duration-matched BBZDR/Wor rats. Endoneurial perfusion was decreased and oxidative stress increased in type 1 BB/Wor rats. C-peptide prevented NCV and neurovascular deficits and attenuated thermal hyperalgesia. Inhibition of nitric oxide (NO) synthase, but not cyclooxygenase, reversed the C-peptide-mediated effects on NCV and nerve blood flow. Indexes of oxidative stress were unaffected by C-peptide. In type 2 BBZDR/Wor rats, neurovascular deficits and increased oxidative stress were unaccompanied by sensory NCV slowing or hyperalgesia. Therefore, nerve oxidative stress is increased and endoneurial perfusion decreased in type 1 BB/Wor and type 2 BBZDR/Wor rats. NO and neurovascular mechanisms, but not oxidative stress, appear to contribute to the effects of C peptide in type 1 EDN. Sensory nerve deficits are not an inevitable consequence of increased oxidative stress and decreased nerve perfusion in a type 2 diabetic rodent model. PMID- 15126238 TI - Synchronization and entrainment of cytoplasmic Ca2+ oscillations in cell clusters prepared from single or multiple mouse pancreatic islets. AB - In contrast to pancreatic islets, isolated beta-cells stimulated by glucose display irregular and asynchronous increases in cytoplasmic Ca(2+) concentration ([Ca(2+)](i)). Here, clusters of 5-30 cells were prepared from a single mouse islet or from pools of islets, loaded with fura-2, and studied with a camera based system. [Ca(2+)](i) oscillations were compared in pairs of clusters by computing the difference in period and a synchronization index lambda. During perifusion with 12 mM glucose, the clusters exhibited regular [Ca(2+)](i) oscillations that were quasi-perfectly synchronized (Delta period of 1.4% and index lambda close to 1.0) between cells of each cluster. In contrast, separate clusters were not synchronized, even when prepared from one single islet. Pairs of clusters neighboring on the same coverslip were not better synchronized than pairs of clusters examined separately (distinct coverslips). We next attempted to synchronize clusters perifused with 12 mM glucose by applying external signals. A single pulse of 20 mM glucose, 10 mM amino acids, or 10 microM tolbutamide transiently altered [Ca(2+)](i) oscillations but did not reset the clusters to oscillate synchronously. On a background of 12 mM glucose, repetitive applications (1 min/5 min) of 10 microM tolbutamide, but not of 20 mM glucose, synchronized separate clusters. Our results identify a level of beta-cell heterogeneity intermediate between single beta-cells and the whole islet. They do not support the idea that substances released by islet cells serve as paracrine synchronizers. However, synchronization can be achieved by an external signal, if this signal has a sufficient strength to overwhelm the intrinsic rhythm of glucose-induced oscillations and is repetitively applied. PMID- 15126239 TI - Mechanical stretch and progesterone differentially regulate activator protein-1 transcription factors in primary rat myometrial smooth muscle cells. AB - During pregnancy, stretch of the uterus, imposed by the growing fetus, is an important signal for the induction of genes involved in the onset of labor. In this study, the expression of activator protein-1 (AP-1) family mRNAs in response to in vitro stretch was investigated in myometrial cells. Rat primary myometrial smooth muscle cells were plated onto collagen I-coated Flex I culture plates and subjected to 25% static stretch on day 4 of culture. Static stretch induced an increase in the expression of c-fos, fosB, fra-1, c-jun, and junB. The expression of both c-fos and junB was maximally induced at 30 min by static stretch. The peak induction for fosB and c-jun occurred at 1 h, whereas the peak of fra-1 induction occurred between 1 and 2 h after application of stretch. Treatment of myometrial cells with progesterone (100 nM, 400 nM, 1 microM) for 1 or 6 h before the application of static stretch did not affect the magnitude of the c-fos response. However, 24 h of progesterone exposure reduced the magnitude of c-fos and fosB stretch induction at both the 400 nM and 1 microM doses. These data indicate that several members of the AP-1 family are stretch-responsive genes in myometrial smooth muscle cells. This response can be attenuated by pretreatment with progesterone; however, the requirement for longer pretreatment times suggests that the inhibitory actions of progesterone do not occur through a direct action of the progesterone receptor within the promoter regions of AP-1 genes. PMID- 15126240 TI - Neutral endopeptidase 24.11 is important for the degradation of both endogenous and exogenous glucagon in anesthetized pigs. AB - Glucagon has a short plasma t(1/2) in vivo, with renal extraction playing a major role in its elimination. Glucagon is degraded by neutral endopeptidase (NEP) 24.11 in vitro, but the physiological relevance of NEP 24.11 in glucagon metabolism is unknown. Therefore, the influence of candoxatril, a selective NEP inhibitor, on plasma levels of endogenous and exogenous glucagon was examined in anesthetized pigs. Candoxatril increased endogenous glucagon concentrations, from 6.3 +/- 2.5 to 20.7 +/- 6.3 pmol/l [COOH-terminal (C)-RIA, P < 0.05]. During glucagon infusion, candoxatril increased the t(1/2) determined by C-RIA (from 3.0 +/- 0.5 to 17.0 +/- 2.5 min, P < 0.005) and midregion (M)-RIA (2.8 +/- 0.5 to 17.0 +/- 3.0 min, P < 0.01) and reduced metabolic clearance rates (MCR; 19.1 +/- 3.2 to 9.4 +/- 2.0 ml.kg(-1).min(-1), P < 0.02, C-RIA; 19.2 +/- 4.8 to 9.0 +/- 2.3 ml.kg(-1).min(-1), P < 0.05, M-RIA). However, neither t(1/2) nor MCR determined by NH2-terminal (N)-RIA were significantly affected (t(1/2), 2.7 +/- 0.4 to 4.5 +/- 1.6 min; MCR, 30.3 +/- 6.4 to 28.5 +/- 9.0 ml.kg(-1).min(-1)), suggesting that candoxatril had no effect on NH2-terminal degradation but leads to the accumulation of NH2-terminally truncated forms of glucagon. Determination of arteriovenous glucagon concentration differences revealed that renal glucagon extraction was reduced (but not eliminated) by candoxatril (from 40.4 +/- 3.8 to 18.6 +/- 4.1%, P < 0.02, C-RIA; 29.2 +/- 3.1 to 14.7 +/- 2.2%, P < 0.02, M-RIA; 26.5 +/- 4.0 to 19.7 +/- 3.5%, P < 0.06, N-RIA). Femoral extraction was reduced by candoxatril when determined by C-RIA (from 22.7 +/- 2.4 to 8.0 +/- 5.1%, P < 0.05) but was not changed significantly when determined using M- or N-RIAs (10.0 +/- 2.8 to 4.7 +/- 3.7%, M-RIA; 10.5 +/- 2.5 to 7.8 +/- 4.2%, N-RIA). This study provides evidence that NEP 24.11 is an important mediator of the degradation of both endogenous and exogenous glucagon in vivo. PMID- 15126241 TI - Leptin treatment markedly increased plasma adiponectin but barely decreased plasma resistin of ob/ob mice. AB - Adiponectin (ApN) and leptin are two adipocytokines that control fuel homeostasis, body weight, and insulin sensitivity. Their interplay is still poorly studied. These hormones are either undetectable or decreased in obese, diabetic ob/ob mice. We examined the effects of leptin treatment on ApN gene expression, protein production, secretion, and circulating levels of ob/ob mice. We also briefly tackled the influence of this treatment on resistin, another adipocytokine involved in obesity-related insulin resistance. Leptin-treated (T) obese mice (continuous sc infusion for 6 days) were compared with untreated lean (L), untreated obese (O), and untreated pair-fed obese (PF) mice. Blood was collected throughout the study. At day 3 or day 6, fat pads were either directly analyzed (mRNA, ApN content) or cultured for up to 24 h (ApN secretion). The direct effect of leptin was also studied in 3T3-F442A adipocytes. Compared with L mice, ApN content of visceral or subcutaneous fat and ApN secretion by adipose explants were blunted in obese mice. Accordingly, plasma ApN levels of O mice were decreased by 50%. Leptin treatment of ob/ob mice increased ApN mRNAs, ApN content, and secretion from the visceral depot by 50-80%. Leptin also directly stimulated ApN mRNAs and secretion from 3T3-F442A adipocytes. After 6 days of treatment, plasma ApN of ob/ob mice increased 2.5-fold, a rise that did not occur in PF mice. Plasma resistin of T mice was barely decreased. Leptin treatment, but not mere calorie restriction, corrects plasma ApN in obese mice by restoring adipose tissue ApN concentrations and secretion, at least in part, via a direct stimulation of ApN gene expression. Such a treatment only minimally affects circulating resistin. ApN restoration could, in concert with leptin, contribute to the metabolic effects classically observed during leptin administration. PMID- 15126242 TI - Pregnancy impairs the counterregulatory response to insulin-induced hypoglycemia in the dog. AB - The impact of pregnancy on the counterregulatory response to insulin-induced hypoglycemia was examined in six nonpregnant (NP) and six pregnant (P; 3rd trimester) conscious dogs by tracer and arteriovenous difference techniques. After basal sampling, insulin was infused intraportally at 30 pmol.kg(-1).min(-1) for 180 min. Insulin rose from 70 +/- 15 to 1,586 +/- 221 pmol/l and 27 +/- 4 to 1,247 +/- 61 pmol/l in the 3rd h in NP and P, respectively. Arterial glucose fell from 5.9 +/- 0.2 to 2.3 +/- 0.2 mmol/l in P. Glucose was infused in NP to equate the rate of fall of glucose and the steady-state concentrations in the groups (5.9 +/- 0.2 to 2.3 +/- 0.1 mmol/l in NP). Glucagon was 32 +/- 6, 69 +/- 11, and 48 +/- 10 ng/l (basal and 1st and 3rd h) in NP, but the response was attenuated in P (34 +/- 5, 46 +/- 6, 41 +/- 9 ng/l). Cortisol and epinephrine rose similarly in both groups, but norepinephrine rose more in NP (Delta3.01 +/- 0.46 and Delta1.31 +/- 0.13 nmol/l, P < 0.05). Net hepatic glucose output (NHGO; micromol.kg(-1).min(-1)) increased from 10.6 +/- 1.8 to 21.2 +/- 3.3 in NP (3rd h) but did not increase in P (15.1 +/- 1.5 to 15.3 +/- 2.8 micromol.kg(-1).min( 1), P < 0.05 between groups). The glycogenolytic contribution to NHGO in NP increased from 5.8 +/- 0.7 to 10.4 +/- 2.5 micromol.kg(-1).min(-1) by 90 min but steadily declined in P. The increase in glycerol levels and the gluconeogenic contribution to NHGO were 50% less in P than in NP, but ketogenesis did not differ. The glucagon and norepinephrine responses to insulin-induced hypoglycemia are blunted in late pregnancy in the dog, impacting on the magnitude of the metabolic responses to the fall in glucose. PMID- 15126243 TI - Discordant effects of a chronic physiological increase in plasma FFA on insulin signaling in healthy subjects with or without a family history of type 2 diabetes. AB - Muscle insulin resistance develops when plasma free fatty acids (FFAs) are acutely increased to supraphysiological levels (approximately 1,500-4,000 micromol/l). However, plasma FFA levels >1,000 micromol/l are rarely observed in humans under usual living conditions, and it is unknown whether insulin action may be impaired during a sustained but physiological FFA increase to levels seen in obesity and type 2 diabetes mellitus (T2DM) (approximately 600-800 micromol/l). It is also unclear whether normal glucose-tolerant subjects with a strong family history of T2DM (FH+) would respond to a low-dose lipid infusion as individuals without any family history of T2DM (CON). To examine these questions, we studied 7 FH+ and 10 CON subjects in whom we infused saline (SAL) or low-dose Liposyn (LIP) for 4 days. On day 4, a euglycemic insulin clamp with [3-3H]glucose and indirect calorimetry was performed to assess glucose turnover, combined with vastus lateralis muscle biopsies to examine insulin signaling. LIP increased plasma FFA approximately 1.5-fold, to levels seen in T2DM. Compared with CON, FH+ were markedly insulin resistant and had severely impaired insulin signaling in response to insulin stimulation. LIP in CON reduced insulin-stimulated glucose disposal (Rd) by 25%, insulin-stimulated insulin receptor tyrosine phosphorylation by 17%, phosphatidylinositol 3-kinase activity associated with insulin receptor substrate-1 by 20%, and insulin-stimulated glycogen synthase fractional velocity over baseline (44 vs. 15%; all P < 0.05). In contrast to CON, a physiological elevation in plasma FFA in FH+ led to no further deterioration in Rd or to any additional impairment of insulin signaling. In conclusion, a 4-day physiological increase in plasma FFA to levels seen in obesity and T2DM impairs insulin action/insulin signaling in CON but does not worsen insulin resistance in FH+. Whether this lack of additional deterioration in insulin signaling in FH+ is due to already well-established lipotoxicity, or to other molecular mechanisms related to insulin resistance that are nearly maximally expressed early in life, remains to be determined. PMID- 15126244 TI - Seeing the trees in the forest: selective electroporation of adipocytes within adipose tissue. AB - Electroporation has been recently adapted for the transfer of macromolecules into cells of tissues in vivo. Although mature adipocytes constitute <20% of cells residing in adipose tissue, we hypothesized that fat cells might be susceptible to selective electrotransfer of plasmid DNA owing to their large size relative to other cells in the tissue. Results demonstrate the feasibility of electroporating DNA into mature fat cells with >99% selectivity over other cells in the tissue. Further experiments used the "adiporation" technique to image the subcellular targeting of fluorescent bioreporter molecules to the nucleus, mitochondria, and lipid droplets of adipocytes within intact adipose tissue. Finally, we utilized fluorescent bioreporters to examine the effects of constitutive activation of the beta-adrenergic signaling pathway in adipocytes. These results demonstrate that overexpression of rat beta1-adrenergic receptors alters the cellular morphology of white adipocytes in a fashion that mimics the effects of systemic infusion of beta3-adrenergic receptor agonists. Hallmarks of the altered morphology include pronounced fragmentation of the single lipid droplet, repositioning of the nucleus, and induction of mitochondrial biogenesis. These results indicate that activation of beta-adrenergic signaling within adipocytes is sufficient to induce a phenotype that resembles typical brown adipocytes and suggest that in vivo electroporation will allow molecular dissection of the mechanisms involved. PMID- 15126245 TI - Fluorescein-methotrexate transport in rat choroid plexus analyzed using confocal microscopy. AB - One function of the vertebrate choroid plexus (CP) is removal of potentially toxic metabolites and xenobiotics from cerebrospinal fluid (CSF) to blood for subsequent excretion in urine and bile. We have used confocal microscopy and quantitative image analysis to follow transport of the large organic anion fluorescein-methotrexate (FL-MTX) from bath (CSF side) to blood vessels in intact rat CP and found concentrative transport from CSF to blood. With 2 microM FL-MTX in the bath, steady-state fluorescence in the subepithelium and vascular spaces exceeded bath levels by 5- to 10-fold, but fluorescence in epithelial cells was below bath levels. FL-MTX accumulation in subepithelium and vascular spaces was reduced by NaCN, Na removal, and by other organic anions, e.g., MTX, probenecid, and estrone sulfate. Increasing medium K 10-fold had no effect. None of these treatments affected cellular accumulation. However, two observations indicated that apical FL-MTX uptake was indeed mediated: first, cellular accumulation was a saturable function of medium substrate concentration; and second, digoxin and MK 571 reduced FL-MTX accumulation in the subepithelial/vascular spaces but also increased cellular accumulation severalfold. In the presence of digoxin and MK 571, cellular accumulation was concentrative, specific, and Na dependent. Thus transepithelial FL-MTX transport involved the following two mediated steps: Na dependent uptake at the apical membrane and electroneutral efflux at the basolateral membrane, possibly on Oatp2 and Mrp1. PMID- 15126246 TI - Water deprivation enhances the inhibitory effect of natriuretic peptides on cAMP synthesis in rat renal glomeruli. AB - This study investigates the effect of water deprivation on the expression of atrial natiruretic peptide (ANP)(1-28) binding sites in rat kidney. Water deprivation increased the B(max) of glomerular binding sites for ANP(1-28) and C type natriuretic peptide (CNP)(1-22) without modifying their affinity, an effect that was prevented in the presence of C-atrial natriuretic factor (C-ANF), suggesting that natriuretic peptide receptor-C (NPR-C) binding sites might be enhanced. Our results indicate that ANP(1-28), CNP(1-22), and C-ANF inhibit cAMP synthesis directly stimulated by forskolin or by the physiological agonists histamine and 5-hydroxytryptamine. The inhibitory effect was found to be significantly greater in water-deprived rats than in controls. Our observations suggest that this effect must be attributed to the 67-kDa NPR-C-like protein, because the 67- and 77-kDa NPR-C-like proteins show high and low affinities for CNP(1-22), respectively, and the enhanced inhibitory effect of CNP on cAMP generation in water-deprived rats was detected at subnanomolar concentrations. In addition, using affinity cross-linking studies we have observed that water deprivation increases the expression of the 67-kDa NPR-C-like protein, and HS 142, which binds to NPR-A and the 77-kDa NPR-C-like but not the 67-kDa protein, reduced ligand internalization without affecting cAMP inhibition by ANP(1-28). Finally, we have found that ligand binding to the 67-kDa NPR-C-like protein is reduced by GTPgammaS, suggesting that this receptor is associated with a G protein in renal glomeruli. The enhanced inhibitory role of natriuretic peptides on cAMP synthesis induced by water deprivation may influence glomerular function in the rat kidney. PMID- 15126247 TI - Functional map of TEA transport activity in isolated rabbit renal proximal tubules. AB - The organic cation (OC) transporters OCT1 and OCT2 are suspected of mediating substrate entry from the blood into proximal tubule cells as the first step in renal secretion of OCs. We examined the contribution of each process in different rabbit renal proximal tubule (RPT) segments, taking advantage of the fact that rabbit orthologs of OCT1 and OCT2 can be distinguished by the high affinity of the former for tyramine (TYR) and of the latter for cimetidine (CIM). We verified that TEA uptake, for which both transporters share a similar affinity, is relatively constant in all three segments (apparent inhibitory constant of 33, 74, and 30 microM and maximal rate of mediated TEA uptake of 0.8, 1.0, and 1.2 pmol x mm(-1) x min(-1) in S1, S2, and S3, respectively). In the S1 segment, TYR was a more effective inhibitor of TEA uptake than CIM (IC50 values of 39 and 328 microM, respectively), implicating OCT1 as the predominant pathway for TEA transport. The opposite profiles were noted in the S2 segment (IC50 values of 302 and 20 microM for TYR and CIM, respectively) and S3 segment (IC50 values of 2,900 and 54 microM for TYR and CIM, respectively), suggesting that OCT2 is the predominant TEA transporter in the later portion of RPT. TEA sufficient to saturate OCT1 and OCT2 blocked only 37% of mediated amantadine transport in the S2 segment, confirming the functional presence of at least one additional OC transporter (perhaps OCT3). These data indicate that renal OC transport involves the concerted activity of a suite of transport processes. PMID- 15126249 TI - Response of descending vasa recta to luminal pressure. AB - We tested whether luminal perfusion and pressurization induce an endothelial cytoplasmic Ca(2+) ([Ca(2+)](CYT)) response in descending vasa recta (DVR). DVR isolated from the rat outer medulla were cannulated and subjected to free-flow microperfusion (5 nl/min); the onset of which increased [Ca(2+)](CYT) from a baseline of 76 +/- 13 to 221 +/- 65 nM. A graded increase in luminal pressure from 0 to 45 mmHg in stopped-flow experiments induced a parallel increase in [Ca(2+)](CYT) from a baseline of 74 +/- 24 to 194 +/- 33 nM at 45 mmHg, with a tendency for [Ca(2+)](CYT) to plateau at pressures >25 mmHg. The removal of extracellular Ca(2+) and blockade by either La(3+) (10 microM) or SKF-96365 (100 microM) eliminated the response. Luminal pressurization to 25 mmHg increased nitric oxide (NO) generation, a response blocked by NO synthase inhibition or removal of extracellular Ca(2+). The NO generation was not affected by the superoxide dismutase mimetic tempol. We conclude that DVR endothelia are mechanosensitive and respond to luminal pressure by elevating [Ca(2+)](CYT) and generating NO. That response might augment medullary perfusion and saliuresis. PMID- 15126248 TI - Megalin mediates renal uptake of heavy metal metallothionein complexes. AB - Although several heavy metal toxins are delivered to the kidney on the carrier protein metallothionein (MT), uncertainty as to how MT enters proximal tubular cells limits treatment strategies. Prompted by reports that MT-I interferes with renal uptake of the megalin ligand beta(2)-microglobulin in conscious rats, we tested the hypothesis that megalin binds MT and mediates its uptake. Three lines of evidence suggest that binding of MT to megalin is critical in renal proximal tubular uptake of MT-bound heavy metals. First, MT binds megalin, but not cubilin, in direct surface plasmon resonance studies. Binding of MT occurs at a single site with a K(d) approximately 10(-4) and, as with other megalin ligands, depends on divalent cations. Second, antisera and various known megalin ligands inhibit the uptake of fluorescently labeled MT in model cell systems. Anti megalin antisera, but not control sera, displace >90% bound MT from rat renal brush-border membranes. Megalin ligands including beta(2)-microglobulin and also recombinant MT fragments compete for uptake by megalin-expressing rat yolk sac BN 16 cells. Third, megalin and fluorescently labeled MT colocalize in BN-16 cells, as shown by fluorescent microscopic techniques. Follow-up surface plasmon resonance and flow cytometry studies using overlapping MT peptides and recombinant MT fragments identify the hinge SCKKSCC region of MT as a critical site for megalin binding. These findings suggest that disruption of the SCKKSCC motif can inhibit proximal tubular MT uptake and thereby eliminate much of the renal accumulation and toxicity of heavy metals such as cadmium, gold, copper, and cisplatinum. PMID- 15126250 TI - AT1A-mediated activation of kidney JNK1 and SMAD2 in obstructive uropathy: preservation of kidney tissue mass using candesartan. AB - Literature suggests the involvement of the renin-angiotensin system and transforming growth factor (TGF)-beta in the renal injury that follows chronic ureteric obstruction. SMAD proteins and the JNK1 cascade are essential components of TGF-beta signaling machinery, and recent data suggest cooperative interaction between JNK1 and SMAD proteins in TGF-beta-mediated gene expression. We used a rat model of chronic unilateral ureteric obstruction to study the effects of candesartan, an AT(1A)-receptor blocker, on tissue morphology and the activities of JNK1 and SMAD2 protein in the kidney. Ureteric obstruction for 28 days leads to interstitial fibrosis, tubule atrophy, and marked activation of SMAD2 and JNK1, without significant change in p38 kinase or ERK. Candesartan treatment, however, attenuated the chronic tubulointerstitial injury in obstructed kidneys and was associated with significant preservation of kidney tissue mass. Furthermore, treatment with candesartan diminished JNK1 activity and downregulated SMAD2 protein and activity in obstructed kidneys. In conclusion, obstructed kidneys showed chronic tubulointerstitial injury, which was associated with JNK1 and SMAD2 activation. The renoprotective effects afforded by AT(1A) receptor blockade in obstructive uropathy are consistent with attenuation of JNK1 and SMAD2-mediated renal injury. PMID- 15126251 TI - Relationship between health status and use of screening mammography and Papanicolaou smears among women older than 70 years of age. AB - BACKGROUND: Older women whose life expectancy is less than 5 years are unlikely to benefit from screening mammography or Papanicolaou (Pap) smears. Since life expectancy is better predicted by health status than by age alone, guidelines recommend considering an older woman's general health when making screening decisions. OBJECTIVE: To determine whether screening mammography and Pap smears are targeted to healthy older women and are avoided in women with limited life expectancies. DESIGN: Cross-sectional population-based study. SETTING: California. PATIENTS: 4792 women 70 years of age or older who participated in the California Health Interview Survey between November 2000 and October 2001. MEASUREMENTS: Predictor variables included age and health status. Health status was measured by using the Medical Outcomes Study 12-item Short Form Physical Summary Scale. The main outcome was self-reported receipt of screening mammography within the previous 2 years and a screening Pap smear within 3 years. RESULTS: Seventy-eight percent of women reported recent screening mammography, and 77% reported a recent Pap smear. Screening rates decreased with advancing age. However, within each age group, the percentage of women reporting screening did not significantly decrease with worsening health status (P > 0.1 for all comparisons). More than half of women 80 years of age or older in the worst health quartile reported recent screening, representing approximately 81,000 mammograms and 35,000 Pap smears. LIMITATIONS: This study relied on self-report, and the interview response rate was 63.7%. CONCLUSIONS: Rates of recent screening mammography and Pap smears are high among older women in California. Although screening rates drop with advancing age, women in poor health do not avoid screening. Screening should be better targeted to healthy older women and should be avoided in women with limited life expectancies for whom risks of screening outweigh potential benefits. PMID- 15126252 TI - Screening for type 2 diabetes mellitus: a cost-effectiveness analysis. AB - BACKGROUND: No randomized, controlled trial of screening for diabetes has been conducted. In the absence of direct evidence, cost-effectiveness models may provide guidance about preferred screening strategies. OBJECTIVE: To estimate the incremental cost-effectiveness of 2 diabetes screening strategies: screening targeted to people with hypertension and universal screening. DESIGN: Markov model. DATA SOURCES: United Kingdom Prospective Diabetes Study, Hypertension Optimal Treatment trial, and recent cost data. TARGET POPULATION: General primary care population in the United States. TIME HORIZON: Lifetime. PERSPECTIVE: Health care system. INTERVENTIONS: Diabetes screening targeted to people with hypertension and universal screening. OUTCOME MEASURES: Cost per quality-adjusted life-year (QALY) gained. Costs (in 1997 U.S. dollars) and QALYs discounted at a 3% annual rate. RESULTS OF BASE-CASE ANALYSIS: At all ages, incremental cost effectiveness ratios were more favorable for screening targeted to people with hypertension than for universal screening. For example, at age 55 years, the cost per QALY for targeted screening compared with no screening was 34,375 dollars, whereas the cost per QALY for universal screening compared with targeted screening was 360,966 dollars. Screening was more cost-effective for ages 55 to 75 years than for younger ages. RESULTS OF SENSITIVITY ANALYSIS: In single-way and probabilistic sensitivity analyses, findings were robust to therapy costs, screening costs, screening lead time, reduced effectiveness of intensive antihypertensive therapy, and increased relative risk reduction for stroke attributable to intensive hypertension control. LIMITATIONS: We did not consider screening targeted to persons with dyslipidemia, and we used studies of people whose diabetes was detected clinically to estimate screening benefits. CONCLUSIONS: Diabetes screening targeted to people with hypertension is more cost effective than universal screening. The most cost-effective strategy is targeted screening at age 55 to 75 years. PMID- 15126253 TI - Coronary vasomotor abnormalities in insulin-resistant individuals. AB - BACKGROUND: Insulin resistance is a metabolic spectrum that progresses from hyperinsulinemia to the metabolic syndrome, impaired glucose tolerance, and finally type 2 diabetes mellitus. It is unclear when vascular abnormalities begin in this spectrum of metabolic effects. OBJECTIVE: To evaluate the association of insulin resistance with the presence and reversibility of coronary vasomotor abnormalities in young adults at low cardiovascular risk. DESIGN: Cross-sectional study followed by prospective, open-label treatment study. SETTING: University hospital. PATIENTS: 50 insulin-resistant and 22 insulin-sensitive, age-matched Mexican-American participants without glucose intolerance or traditional risk factors for or evidence of coronary artery disease. INTERVENTION: 3 months of thiazolidinedione therapy for 25 insulin-resistant patients. MEASUREMENTS: Glucose infusion rate in response to insulin infusion was used to define insulin resistance (glucose infusion rate < or = 4.00 mg/kg of body weight per minute [range, 0.90 to 3.96 mg/kg per minute]) and insulin sensitivity (glucose infusion rate > or = 7.50 mg/kg per minute [range, 7.52 to 13.92 mg/kg per minute]). Myocardial blood flow was measured by using positron emission tomography at rest, during cold pressor test (largely endothelium-dependent), and after dipyridamole administration (largely vascular smooth muscle-dependent). RESULTS: Myocardial blood flow responses to dipyridamole were similar in the insulin-sensitive and insulin-resistant groups. However, myocardial blood flow response to cold pressor test increased by 47.6% from resting values in insulin-sensitive patients and by 14.4% in insulin-resistant patients. During thiazolidinedione therapy in a subgroup of insulin-resistant patients, insulin sensitivity improved, fasting plasma insulin levels decreased, and myocardial blood flow responses to cold pressor test normalized. LIMITATIONS: The study was not randomized, and it included only 1 ethnic group. CONCLUSIONS: Insulin-resistant patients who do not have hypercholesterolemia or hypertension and do not smoke manifest coronary vasomotor abnormalities. Insulin-sensitizing thiazolidinedione therapy normalized these abnormalities. These results suggest an association between insulin resistance and abnormal coronary vasomotor function, a relationship that requires confirmation in larger studies. PMID- 15126254 TI - Early diagnosis of subclinical multidrug-resistant tuberculosis. AB - BACKGROUND: Tuberculosis control hinges on prompt diagnosis of active cases and screening of contacts by tuberculin skin testing. Rapid blood tests for Mycobacterium tuberculosis infection are a new alternative to the tuberculin skin test, but whether they improve clinical outcomes is unknown. OBJECTIVE: To describe how a novel T-cell-based test for M. tuberculosis infection helped diagnose tuberculosis in an asymptomatic, immunosuppressed adult with a negative result on a tuberculin skin test. DESIGN: Case report. SETTING: Household contact. PATIENTS: Asymptomatic man receiving maintenance azathioprine therapy for Crohn disease whose wife had multidrug-resistant pulmonary tuberculosis. MEASUREMENTS: Enzyme-linked immunospot (ELISPOT) assay, computed tomography, and bronchoalveolar lavage cultures. RESULTS: The man had a negative tuberculin skin test result and a positive ELISPOT assay result. High-resolution computed tomography of the chest showed consolidation with early cavitation. Bronchoalveolar lavage and culture confirmed multidrug-resistant tuberculosis. LIMITATIONS: This single case report is a proof of concept and is not a formal evaluation of clinical utility. CONCLUSIONS: A positive ELISPOT assay result helped diagnose subclinical active tuberculosis in an immunosuppressed patient with a false-negative tuberculin skin test result. Large prospective studies that compare benefits and costs of this alternative to tuberculin skin testing are needed. PMID- 15126255 TI - The quality of pharmacologic care for vulnerable older patients. AB - BACKGROUND: Although pharmacotherapy is critical to the medical care of older patients, medications can have considerable toxicity in this age group. To date, research has focused on inappropriate prescribing and policy efforts have aimed at access, but no comprehensive measurement of the quality of pharmacologic management using explicit criteria has been performed. OBJECTIVE: To evaluate the broad range of pharmacologic care processes for vulnerable older patients. DESIGN: Observational cohort study. SETTING: 2 managed care organizations enrolling older persons. PATIENTS: Community-dwelling high-risk patients 65 years of age or older continuously enrolled in the managed care organizations from 1 July 1998 to 31 July 1999. MEASUREMENTS: Patients' receipt of care as specified in 43 quality indicators covering 4 domains of pharmacologic care: 1) prescribing indicated medications; 2) avoiding inappropriate medications; 3) education, continuity, and documentation; and 4) medication monitoring. RESULTS: Of 475 vulnerable older patients, 372 (78%) consented to participate and had medical records that could be abstracted. The percentage of appropriate pharmacologic management ranged from 10% for documentation of risks of nonsteroidal anti inflammatory drugs to 100% for avoiding short-acting calcium-channel blockers in patients with heart failure and avoiding beta-blockers in patients with asthma. Pass rates for quality indicators in the "avoiding inappropriate medications" domain (97% [95% CI, 96% to 98%]) were significantly higher than pass rates for "prescribing indicated medications" (50% [CI, 45% to 55%]); "education, continuity, and documentation" (81% [CI, 79% to 84%]); and "medication monitoring" (64% [CI, 60% to 68%]). LIMITATIONS: Fewer than 10 patients were eligible for many of the quality indicators measured, and the generalizability of these findings in 2 managed care organizations to the general geriatric population is uncertain. CONCLUSIONS: Failures to prescribe indicated medications, monitor medications appropriately, document necessary information, educate patients, and maintain continuity are more common prescribing problems than use of inappropriate drugs in older patients. PMID- 15126256 TI - Patient education materials about the treatment of early-stage prostate cancer: a critical review. AB - BACKGROUND: To ensure that patients make informed medical decisions, patient education materials must communicate treatment risks and benefits. OBJECTIVE: To survey publicly available patient education materials and assess their suitability to support informed decision making in early-stage prostate cancer. DESIGN: Cross-sectional review of Internet, print, and multimedia sources. SETTING: University data analysis laboratory. MEASUREMENTS: The content of 44 materials that described all standard treatment options was reviewed. Top-rated documents underwent plain-language review. Total score on 54 content items and accuracy, balance, and plain-language evaluation was measured. RESULTS: 502 of 546 patient education materials did not describe all standard treatments (watchful waiting, surgery, radiation, and hormone therapy). Eighty percent of the 44 materials that addressed standard treatments and underwent content review described anatomy, physiology, stage, and grade of cancer. Half of the materials fully described radical prostatectomy and radiation therapy. One third of the materials included risks and benefits of each treatment; none explicitly compared outcomes of all treatments in a single summary. Information was accurate and balanced but did not include key content for informed consent. LIMITATIONS: The search was restricted to publicly available materials and did not include books or materials written in languages other than English. The accuracy, balance, and plain-language reviews were evaluated by 1 reviewer. The criteria reflect the authors' focus on informed decision making. Other aspects of health education may require a different evaluation template. CONCLUSIONS: Currently available patient education materials on early-stage prostate cancer treatment do not contain comprehensive information about the risks and benefits of each treatment. To assist patients and physicians in choosing among prostate cancer treatment options, a new generation of materials is needed. PMID- 15126257 TI - Stem-cell transplantation in myocardial infarction: a status report. AB - Myocardial infarction is the leading cause of congestive heart failure and death in the industrialized world. Current therapy is limited in preventing the progression of ventricular remodeling and congestive heart failure. Recent interest has focused on stem cells, which are undifferentiated and pluripotent cells that can proliferate, potentially self-renew, and differentiate into cardiomyocytes. Myocardial regeneration with stem-cell transplantation is a possible treatment option to reverse the deleterious hemodynamic and neurohormonal effects that occur after myocardial infarction and can lead to congestive heart failure. Various preclinical animal studies show the potential to regenerate myocardium and improve perfusion to the infarct area to improve cardiac function but also suggest that stem cells may have proarrhythmic effects. Early phase I clinical studies indicate that stem-cell transplantation is feasible and may have beneficial effects on ventricular remodeling after myocardial infarction. Future randomized clinical trials will establish the magnitude of the benefit and the effects on arrhythmias after stem-cell therapy. PMID- 15126258 TI - Lung cancer screening: recommendation statement. AB - This statement summarizes the current U.S. Preventive Services Task Force (USPSTF) recommendation on screening for lung cancer and the supporting scientific evidence and updates the 1996 recommendations on this topic. In 1996, the USPSTF recommended against screening for lung cancer (a grade D recommendation). The Task Force now uses an explicit process in which the balance of benefits and harms is determined exclusively by the quality and magnitude of the evidence. As a result, current letter grades are based on different criteria than those used in 1996. The complete information on which this statement is based, including evidence tables and references, is available in the accompanying article in this issue and in the systematic evidence review on this topic, available through the USPSTF Web site (http://www.preventiveservices.ahrq.gov) and the National Guideline Clearinghouse (http://www.guideline.gov). The complete USPSTF recommendation statement (which includes a brief review of the supporting evidence) and the summary of the evidence are also available in print through the Agency for Healthcare Research and Quality Publications Clearinghouse (telephone, 800-358-9295; e-mail, ahrqpubs@ahrq.gov). PMID- 15126259 TI - Lung cancer screening with sputum cytologic examination, chest radiography, and computed tomography: an update for the U.S. Preventive Services Task Force. AB - BACKGROUND: Lung cancer is the leading cause of cancer-related death in the United States and worldwide. No major professional organizations, including the U.S. Preventive Services Task Force (USPSTF), currently recommend screening for lung cancer. PURPOSE: To examine the evidence evaluating screening for lung cancer with chest radiography, sputum cytologic examination, and low-dose computed tomography (CT) to aid the USPSTF in updating its recommendation on lung cancer screening. DATA SOURCES: MEDLINE, the Cochrane Library, reviews, editorials, and experts. STUDY SELECTION: Studies that evaluated mass screening programs for lung cancer involving the tests of interest were selected. All studies were reviewed, but only studies with control groups were rated in quality since these would most directly influence the USPSTF screening recommendation. DATA EXTRACTION: Data were abstracted to data collection forms. Studies were graded according to criteria developed by the USPSTF. DATA SYNTHESIS: None of the 6 randomized trials of screening for lung cancer with chest radiography alone or in combination with sputum cytologic examination showed benefit among those screened. All studies were limited because some level of screening occurred in the control population. Five case-control studies from Japan suggested benefit to both high- and low-risk men and women. All studies were limited by potential healthy screenee bias. Six cohort studies showed that when CT was used to screen for lung cancer, lung cancer was diagnosed at an earlier stage than in usual clinical care. However, these studies did not have control groups, making mortality evaluation difficult. In addition, the studies demonstrated a high rate of false-positive findings. CONCLUSIONS: Current data do not support screening for lung cancer with any method. These data, however, are also insufficient to conclude that screening does not work, particularly in women. Two randomized trials of screening with chest radiography or low-dose CT are currently under way and will better inform lung cancer screening decisions. PMID- 15126260 TI - Right and wrong reasons to be screened. PMID- 15126261 TI - Screening for diabetes: can we afford not to screen? PMID- 15126262 TI - Sara and George and justice. PMID- 15126263 TI - Cost-effectiveness of cyclooxygenase-2 inhibitors in chronic arthritis. PMID- 15126264 TI - Diagnosis and management of adults with pharyngitis. PMID- 15126265 TI - Diagnosis and management of adults with pharyngitis. PMID- 15126266 TI - The discrepancy between observational studies and randomized trials of menopausal hormone therapy. PMID- 15126267 TI - Mammography and palpable breast abnormalities. PMID- 15126268 TI - Helicobacter pylori and idiopathic thrombocytopenic purpura. PMID- 15126269 TI - Summaries for patients. Relationship of health status and use of mammography and Papanicolaou smears among women 70 years of age or older. PMID- 15126270 TI - Summaries for patients. The cost-effectiveness of screening for type 2 diabetes. PMID- 15126271 TI - Summaries for patients. Insulin resistance is associated with abnormalities of coronary arteries in people without other cardiac risk factors. PMID- 15126272 TI - Summaries for patients. The quality of pharmacologic care for older adults in two managed care organizations. PMID- 15126273 TI - Summaries for patients. Can stem cells restore cardiac tissue after a heart attack? PMID- 15126274 TI - Summaries for patients. Screening for lung cancer: recommendations from the U.S. Preventive Services Task Force. PMID- 15126275 TI - Delayed drug hypersensitivity reactions. PMID- 15126277 TI - Mitochondrial pathogenesis from genes and apoptosis to aging and disease. Proceedings of the First Scientific Meeting of the Asian Society for Mitochondrial Research and Medicine. February 5-6, 2003. Seoul, Korea. PMID- 15126278 TI - Mitochondrial pathogenesis from genes and apoptosis to aging and disease. Overview. PMID- 15126279 TI - Mitochondrial genome single nucleotide polymorphisms and their phenotypes in the Japanese. AB - Polymorphisms in the human mitochondrial genome have been used for the elucidation of phylogenetic relationships among various ethnic groups. Because analysis by mitochondrial genetics has detected pathogenic mutations causing mitochondrial encephalomyopathy or cardiomyopathy, most of the mitochondrial single nucleotide polymorphisms (mtSNPs) found in control subjects have been regarded as merely normal variants. However, we cannot exclude the possibility that the mitochondrial functional differences among individuals are ascribable at least in part to the mtSNPs of each individual. Human lifespan in ancient history was much shorter than that at the present time. Therefore, it is reasonable to speculate that certain mtSNPs that predispose one toward susceptibility to adult- or elderly-onset diseases, such as Parkinson's disease and Alzheimer's disease, have never been a target for natural selection in the past. Similarly, thrifty mtSNPs that had been advantageous for survival under severe famine or cold climate conditions might turn out to be related to satiation-related diseases, such as diabetes mellitus and obesity. To examine these hypotheses, we have constructed a mtSNP database by sequencing the entire mitochondrial genomes of 672 subjects: 96 in each of seven groups (i.e., centenarians, young obese or non obese subjects, diabetic patients with or without major vascular involvement, patients with Parkinson's disease, and those with Alzheimer's disease). PMID- 15126280 TI - Mitochondrial DNA variation in the aboriginal populations of the Altai-Baikal region: implications for the genetic history of North Asia and America. AB - The discovery of mtDNA types common to Asians and Amerinds (types A, B, C, and D) forced investigators to search for those nations of Asia which, though not considered the ancestors of the Amerinds, have retained a close genetic resemblance with them. We collected samples and studied the gene pools of the Turkic-speaking nations of South Siberia: Altaians, Khakassians, Shorians, Tuvinians, Todjins, Tofalars, Sojots, as well as Mongolian-speaking Buryats. The data indicate that nearly all Turkic-speaking nations of Siberia and Central Asia, as well as the Buryats, have types A, B, C, and D in their gene pool. The highest total frequency of these types is observed in the Tuvinians and Sojots. They, as well as the Buryats, also have the lowest frequency of the europeoid types. The most mixed Asian-Europeoid gene pool examined turned out to be that of the Shorians. An important finding was the presence of type X in the Altaians, which had not yet been detected in Asia. As shown by computer analysis, this DNA sequence is not a late European admixture. Rather, the Altai variant X is ancient and can be close to the ancestral form of the variants of contemporary Europeans and Amerinds. The presented results prove that of all nations in Asia, the Turkic speaking nations living between Altai and Baikal along the Sayan mountains are genetically closest to the Amerinds. PMID- 15126281 TI - Mitochondrial ALDH2 deficiency as an oxidative stress. AB - Mitochondrial aldehyde dehydrogenase 2 (ALDH2) plays a major role in ethanol metabolism. It is involved in acetaldehyde detoxification. A polymorphism of the ALDH2 gene is specific to North-East Asians. Sensitivity to ethanol is highly associated with this polymorphism (ALDH2(*)2 allele), which is responsible for a deficiency of ALDH2 activity. We first show that this deficiency influences the risk for late-onset Alzheimer's disease (LOAD) by a case-control study in a Japanese population. In a comparison of 447 patients with sex, age, and region matched non-demented controls, the genotype frequency for the ALDH2(*)2 allele was significantly higher in the patients than in the controls (P=0.001). Next, we examined the combined effect of the ALDH2(*)2 and the apolipoprotein E4 allele (APOE-epsilon4), which has been confirmed to be a risk factor for LOAD. The ALDH2(*)2 allele more significantly affected frequency and age at onset in patients with APOE-epsilon4 than in those without it. These results indicate that the ALDH2 deficiency is a risk factor for LOAD, acting synergistically with the APOE-epsilon allele. Next, to elucidate the molecular mechanism involved, we obtained ALDH2-deficient cell lines by introducing mouse mutant ALDH2 cDNA into PC12 cells. We speculate that ALDH2 may act to oxidize toxic aldehyde derivatives. Then, we found that the ALDH2-deficient transfectants were highly vulnerable to exogenous 4-hydroxy-2-nonenal, an aldehyde derivative generated from peroxidized fatty acids. In addition, the ALDH2-deficient transfectants were sensitive to oxidative insult induced by antimycin A, accompanied by an accumulation of proteins modified with 4-hydroxy-2-nonenal. Mitochondrial ALDH2 functions as a protector against oxidative stress. PMID- 15126282 TI - Mitochondrial reactive oxygen species generation and calcium increase induced by visible light in astrocytes. AB - Mitochondria contain photosensitive chromophores that can be activated or inhibited by light in the visible range. Rather than utilizing light energy, however, mitochondrial electron transport oxidation-reduction reaction and energy coupling could be stimulated or damaged by visible light. Our previous work demonstrated that reactive oxygen species (ROS) were generated in cultured astrocytes after visible laser irradiation. With confocal fluorescence microscopy, we found that ROS were generated mostly from mitochondria. This mitochondrial ROS (mROS) formation plays a critical role in photoirradiation induced phototoxicity and apoptosis. In this study, we measured changes of mitochondrial calcium level ([Ca(2+)](m)) in cultured astrocytes (RBA-1 cell line) irradiated with blue light and examined the association between mROS formation and [Ca(2+)](m) level changes. Changes of intracellular ROS and [Ca(2+)](m) were visualized using fluorescent probes 2',7' dichlorodihydrofluorescein (DCF), and rhod-2. After exposure to visible light irradiation, RBA-1 astrocytes showed a rapid increase in ROS accumulation particularly in the mitochondrial area. Increase in [Ca(2+)](m) was also induced by photoirradiation. The levels of increase in DCF fluorescence intensity varied among different astrocytes. Some of the cells generated much higher levels of ROS than others. For those cells that had high ROS levels, mitochondrial Ca(2+) levels were also high. In cells that had mild ROS levels, mitochondrial Ca(2+) levels were only slightly increased. The rate of increase in DCF fluorescence seemed to be close to the rate of rhod-2 fluorescence increase. There is a positive and close correlation between mitochondrial ROS levels and mitochondrial Ca(2+) levels in astrocytes irradiated by visible light. PMID- 15126283 TI - Radical metabolism is partner to energy metabolism in mitochondria. AB - It has been shown that cytochrome c plays a role in scavenging O(2)(-). and H(2)O(2) in mitochondria through two electron leak pathways of the respiratory chain. Based on the two electron leak pathways and the superoxide metabolic routes in nature, it is suggested that the concept of radical metabolism should be added as a partner to the energy metabolism of mitochondria. A total of four reactive pathways of superoxide anion have been identified, collected, and linked to electron leaks in the respiratory chain. A view is presented that envisions mitochondria working on ATP synthesis by means of transferring electrons inside the respiratory chain while at the same time working on ROS generation and elimination as electrons leak out of the chain. PMID- 15126284 TI - Mitochondrial nucleoid and transcription factor A. AB - Nuclear DNA is tightly packed into nucleosomal structure. In contrast, human mitochondrial DNA (mtDNA) had long been believed to be rather naked because mitochondria lack histone. Mitochondrial transcription factor A (TFAM), a member of a high mobility group (HMG) protein family and a first-identified mitochondrial transcription factor, is essential for maintenance of mitochondrial DNA. Abf2, a yeast counterpart of human TFAM, is abundant enough to cover the whole region of mtDNA and to play a histone-like role in mitochondria. Human TFAM is indeed as abundant as Abf2, suggesting that TFAM also has a histone-like architectural role for maintenance of mtDNA. When human mitochondria are solubilized with non-ionic detergent Nonidet-P40 and then separated into soluble and particulate fractions, most TFAM is recovered from the particulate fraction together with mtDNA, suggesting that human mtDNA forms a nucleoid structure. TFAM is tightly associated with mtDNA as a main component of the nucleoid. PMID- 15126285 TI - Regulation of mitochondrial transcription factor A expression by high glucose. AB - Mitochondrial transcription factor A (Tfam, previously mtTFA) is a key regulator of mitochondrial DNA (mtDNA) transcription and replication. We have reported that overexpression of nuclear respiratory factor-1 (NRF-1) and high concentration (50 mM) of glucose increased the promoter activity of the rat Tfam in L6 rat skeletal muscle cells. In this study, we investigated the mechanism of high glucose induced Tfam transactivation. The addition of 50 mM glucose for 24 h increased Tfam promoter activity up to twofold. The glucose-induced Tfam expression was dose-dependent and cell-type specific. Glucose increased the Tfam promoter-driven transactivity in L6 (skeletal muscle), HIT (pancreatic beta-cell), and CHO (ovary) cells, but not in HepG2 (hepatoma), HeLa, and CV1 (kidney) cells. Among various monosaccharides, only glucose and fructose increased the Tfam promoter activity. Oxidative stress might not be involved in glucose-induced Tfam expression since treatment with antioxidants such as vitamin C, vitamin E, probucol, or alpha-lipoic acid did not suppress the induction. None of the inhibitors of protein kinase C, MAP kinase, and PI3 kinase altered the glucose induced Tfam promoter activity, suggesting that general phosphorylation is involved in its signaling. However, a dominant negative mutant of NRF-1, in which 200 amino acids of C-terminus were truncated, completely suppressed the glucose induced Tfam induction. It was concluded that high glucose-induced Tfam transcription in L6 cells might be mediated by NRF-1. PMID- 15126286 TI - Regulation and role of the mitochondrial transcription factor in the diabetic rat heart. AB - To clarify the mechanism of abnormalities in mitochondrial expression and function in diabetic rat heart, we have studied the transcriptional activities of mitochondrial DNA using isolated intact mitochondria from the heart of either diabetic or control rats. The transcriptional activity of cardiac mitochondria isolated from diabetic rats decreased to 40% of the control level (P < 0.01). Consistently, in the heart of diabetic rats, the content of cytochrome b mRNA encoded by mitochondrial DNA was reduced to 50% of control (P < 0.01). This abnormal transcriptional activity of mitochondrial DNA could not be explained by mRNA or protein contents of mitochondrial transcription factor (mtTFA), but mtTFA binding to the promoter sequence of mitochondrial DNA, assessed by gel-shift assay, was attenuated in diabetic rats. In contrast, the mRNA expression of nuclear-encoded mitochondrial genes, such as ATP synthase-beta, was not affected by diabetes. Although O(2) consumption of the mitochondria from diabetic rats was decreased, H(2)O(2) production in these rats was increased compared with the control. Insulin treatment reversed all the abnormalities found in diabetic rats. These results clearly indicate that an impairment of binding activity of mtTFA to the promoter sequence has a key role in the abnormal mitochondrial gene expression, which might explain the mitochondrial dysfunction found in diabetic heart. PMID- 15126287 TI - The mitochondrial production of reactive oxygen species in relation to aging and pathology. AB - Mitochondria are known to be strong producers of reactive oxygen species (ROS) and, at the same time, particularly susceptible to the oxidative damage produced by their action on lipids, proteins, and DNA. In particular, damage to mtDNA induces alterations to the polypeptides encoded by mtDNA in the respiratory complexes, with consequent decrease of electron transfer, leading to further production of ROS and thus establishing a vicious circle of oxidative stress and energetic decline. This deficiency in mitochondrial energetic capacity is considered the cause of aging and age-related degenerative diseases. Complex I would be the enzyme most affected by ROS, since it contains seven of the 13 subunits encoded by mtDNA. Accordingly, we found that complex I activity is significantly affected by aging in rat brain and liver mitochondria as well as in human platelets. Moreover, due to its rate control over aerobic respiration, such alterations are reflected on the entire oxidative phosphorylation system. We also investigated the role of mitochondrial complex I in superoxide production and found that the one-electron donor to oxygen is most probably the Fe-S cluster N2. Short chain coenzyme Q (CoQ) analogues enhance ROS formation, presumably by mediating electron transfer from N2 to oxygen, both in bovine heart SMP and in cultured HL60 cells. Nevertheless, we have accumulated much evidence of the antioxidant role of reduced CoQ(10) in several cellular systems and demonstrated the importance of DT-diaphorase and other internal cellular reductases to reduce exogenous CoQ(10) after incorporation. PMID- 15126288 TI - Biological significance of the defense mechanisms against oxidative damage in nucleic acids caused by reactive oxygen species: from mitochondria to nuclei. AB - In mammalian cells, more than one genome in a single cell has to be maintained throughout the entire life of the cell, namely, one in the nucleus and the other in the mitochondria. The genomes and their precursor nucleotides are highly exposed to reactive oxygen species, which are inevitably generated as a result of the respiratory function in mitochondria. To counteract such oxidative damage in nucleic acids, cells are equipped with several defense mechanisms. Modified nucleotides in the nucleotide pools are hydrolyzed, thus avoiding their incorporation into DNA or RNA. Damaged bases in DNA with relatively small chemical alterations are mainly repaired by the base excision repair (BER) system, which is initiated by the excision of damaged bases by specific DNA glycosylases. MTH1 protein hydrolyzes oxidized purine nucleoside triphosphates, such as 8-oxo-dGTP, 8-oxo-dATP, and 2-hydroxy (OH)-dATP to the monophosphates, and MTH1 are located in the cytoplasm, mitochondria, and nucleus. We observed an increased susceptibility to spontaneous carcinogenesis in Mth1-deficient mice and an alteration of MTH1 expression along with the accumulation of 8-oxo-dG in patients with various neurodegenerative diseases. Enzymes for the BER pathway, namely, 8-oxoG DNA glycosylase (OGG1), 2-OH-A/adenine DNA glycosylase (MUTYH), and AP endonuclease (APEX2) are also located both in the mitochondria and in the nuclei, and the expression of mitochondrial OGG1 is altered in patients with various neurodegenerative diseases. We also observed increased susceptibilities to spontaneous carcinogenesis in OGG1 and MUTYH-deficient mice. The increased occurrence of lung tumor in OGG1-deficient mice was completely abolished by the concomitant disruption of the Mth1 gene. PMID- 15126289 TI - Mitochondrial swelling and generation of reactive oxygen species induced by photoirradiation are heterogeneously distributed. AB - Abundant evidence has been gathered to show that overproduction of reactive oxygen species (ROS) can lead to the opening of the mitochondrial permeability transition pore (MPTP) and result in apoptosis in mammalian cells. The information regarding spatial and temporal regulation of intracellular ROS formation related to the MPTP opening, however, is relatively limited. In this study, we used a fluorescent probe, dihydro-2',7'-dichloroforescin (DCF), to detect intracellular ROS levels in different compartments of the cell in a time resolved manner. The roles of mitochondrial ROS (mROS) in the MPTP opening and mitochondrial membrane potential drop were investigated by using H(2)DCFDA coloaded with a mitochondrial marker dye MitoTracker Red, and by a mitochondrial membrane potential dye tetramethyl rhodamine ethyl ester. We applied multiphoton laser scanning microscopy to avoid autooxidation and bleaching of DCF so that long-term visualization of intracellular ROS formation could be performed. Moreover, we noted that the resting mROS levels of different mitochondria were not homogeneous. After cells had been exposed to photoirradiation, the intracellular ROS gradually increased but the heterogeneity of mROS was maintained. Later, swelling was observed in mitochondria that contained higher levels of ROS, indicating the opening of the MPTP. In cells in which all the mitochondria swelled, they were translocated to the perinuclear area, which became the site of ROS production. At this stage, mROS reached the highest level concomitantly with a complete loss of mitochondrial membrane potential, indicating full opening of the MPTP. At the end, photoirradiation resulted in apoptotic cell death. In summary, we demonstrated by multiphoton laser scanning microscopy that photoirradiation induces heterogeneous intracellular ROS formation and mitochondrial permeability transition pore opening in single intact cells. These observations imply the existence of a microdomain in the regulation of mROS formation and subsequent opening of the MPTP. PMID- 15126290 TI - Mitochondrial dysfunction via disruption of complex II activity during iron chelation-induced senescence-like growth arrest of Chang cells. AB - When cells are deprived of iron, their growth is invariably inhibited. However, the mechanism involved remains largely unclear. Recently, we have reported that subcytotoxic concentration of deferoxamine mesylate (DFO), an iron chelator, specifically inhibited transition of Chang cell, a normal hepatocyte cell line, from G1 to S phase, which was accompanied by irreversible appearance of senescent biomarkers. To investigate factors responsible for the irreversible arrest, we examined mitochondrial activities because they require several irons for their proper structure and function. After exposure to 1 M DFO, total cellular ATP level was irreversibly decreased with concurrent disruption of mitochondrial membrane potential (DeltaPsim), implying that it might be one of the crucial factors involved in the arrest. DFO did not directly inhibit the mitochondrial respiratory activities in vitro. Among the respiratory activities, complex II activity was specifically inhibited through a down-regulation of the expression of its iron-sulfur subunit. We also observed that mitochondrial morphology was drastically changed to highly elongated form. Our results suggest that mitochondrial function is sensitive to cellular iron level and iron deprivation might be involved in inducing the senescent arrest. In addition, complex II, which is a part of both oxidative phosphorylation and the Krebs cycle, could be one of the critical factors that regulate mitochondrial function by responding to iron levels. PMID- 15126291 TI - Mitochondrial DNA mutation and depletion increase the susceptibility of human cells to apoptosis. AB - Mitochondrial diseases, such as MELAS, MERRF, and CPEO syndromes, are associated with specific point mutations or large-scale deletions of mitochondrial DNA (mtDNA), which impair mitochondrial respiratory functions and result in decreased production of ATP in affected tissues. Recently, mitochondria have been recognized to act as key players in the regulation of cell death. To investigate whether a pathogenic mutation of mtDNA exerts any effect on the process of apoptosis of human cells, we constructed a series of cybrid human cells harboring different proportions of mtDNA with the A3243G or the A8344G transition, or with the 4,977-bp deletion, by cytoplasmic fusion of patients' skin fibroblasts with mtDNA-depleted rho(0) cells of an immortal human osteosarcoma cell line (143B). We observed that the decrease in cell viability upon staurosporine treatment or exposure to ultraviolet (UV) irradiation was more pronounced in the cybrids harboring high levels of mutated mtDNA compared with the control cybrids. Using DNA fragmentation analysis, we found that the cell death induced by treatment with 100 nM staurosporine or by exposure to UV irradiation at 20 J/m(2) was caused by apoptosis, not necrosis. Moreover, we demonstrated activation of caspase 3 by Western blot and enhanced release of cytochrome c after 100 nM staurosporine treatment or 20 J/m(2) UV irradiation of the cybrids harboring high levels of the three mtDNA mutations. Furthermore, as compared with parental osteosarcoma 143B cells, the rho(0) cells were found to be more susceptible to apoptosis, which was accompanied by caspase 3 activation and cytochrome c release. This indicates that mtDNA plays an important role in the regulation of apoptosis in human cells. Taken together, these findings suggest that mutation and depletion of mtDNA increase the susceptibility of human cells to apoptosis triggered by exogenous stimuli such as UV irradiation or staurosporine. PMID- 15126292 TI - Resistance of rho(0) cells against apoptosis. AB - Mitochondrion is one of the master players in both apoptosis and necrosis. However, most previous articles report that mitochondrial DNA-depleted cells without oxidative phosphorylation underwent apoptosis by several apoptotic effectors as efficiently as their parental cells, suggesting that intact mitochondrial function is dispensable for the progression of apoptosis. We studied the role of mitochondrial function in several apoptosis models. TRAIL, a recently identified member of the TNF family with cytotoxicity on a wide variety of transformed cells, killed SK-Hep1 cells with characteristic features of apoptosis such as DNA fragmentation, sub-G1 ploidy peak, and cytochrome c translocation. In contrast with parental cells, mitochondrial DNA-deficient SK Hep1 rho(0) cells were resistant to TRAIL-induced apoptosis. Dissipation of mitochondrial potential or cytochrome c translocation did not occur in rho(0) cells after TRAIL treatment. Bax translocation also was absent in rho(0) cells, accounting for the failure of cytochrome c release in rho(0) cells. SK-Hep1 rho(0) cells were resistant to other death effectors such as staurosporine. Our results indicate that apoptosis of SK-Hep1 hepatoma cells is dependent on intact mitochondrial function. Because aged cells or tumor cells have frequent mutations or deletions of mitochondrial DNA, they might acquire the ability to evade apoptosis or tumor surveillance imposed by TRAIL or other death effectors in vivo, accounting for the selection advantage of cancer cells and frequent development of cancer in aged individuals. PMID- 15126293 TI - Mitochondrial DNA 4,977-bp deletion in paired oral cancer and precancerous lesions revealed by laser microdissection and real-time quantitative PCR. AB - Oral cancer is the fourth leading cause of cancer deaths among men in Taiwan and is closely associated with areca quid chewing habits. Recent studies showed that mitochondrial DNA (mtDNA) mutations occur in various tumors, including oral cancers, and that the accumulation of mtDNA deletions could be an important contributor to carcinogenesis. Using laser microdissection, we have analyzed mtDNA deletions by pairwise comparisons in oral cancer, precancerous cells, and their adjacent submucosal stoma tissues in 12 patients with areca quid chewing history. Real-time quantitative polymerase chain reaction (RTQPCR) was performed to detect and quantify mtDNA with the 4,977-bp deletion in the histologically defined specified cell groups. Quantitative analysis of 60 samples by RTQPCR revealed that the average proportions of 4,977-bp deleted mtDNA over total mtDNA were 0.137%, 0.367%, and 0.001% in cancer, precancer cells, and lymphocytes of lymph node biopsies, respectively. Pairwise analysis of the proportion of mtDNA deletion in cancer, precancer, and their stroma tissues revealed a consistent trend among these patients. All of the patients (12/12) presented a higher proportion of mtDNA with 4,977-bp deletion in the lesions than in the lymphocytes, with average increases of 198-fold in cancer and 546-fold in precancer cells. A decrease in the proportion of deleted mtDNA was observed in 8 of 12 patients when the disease progressed from precancer to cancer lesions. Interestingly, 7 of 12 cancer tissues and 8 of 12 precancer lesions exhibited an average of 6.3-fold and 17.4-fold increases in the proportion of 4,977-bp deleted mtDNA in the stromal cells than in the lesion cells, respectively. The observation that the proportion of 4,977-bp deleted mtDNA in all oral lesions was higher than normal and consistently decreased during cancer progression from precancer to primary cancer suggests that accumulation and subsequent cytoplasmic segregation of the mutant mtDNA during cell division may play an important role in oral carcinogenesis. This study also demonstrates that laser microdissection combined with RTQPCR is an efficient and sensitive tool to gain insight into the role that mtDNA mutation may play in carcinogenesis. PMID- 15126294 TI - Role of oxidative stress in pancreatic beta-cell dysfunction. AB - Oxidative stress is produced under diabetic conditions and is likely involved in progression of pancreatic beta-cell dysfunction found in diabetes. Possibly caused by low levels of antioxidant enzyme expressions, pancreatic beta-cells are vulnerable to oxidative stress. When beta-cell-derived HIT-T15 cells or isolated rat islets were exposed to oxidative stress, insulin gene expression was markedly decreased. To investigate the significance of oxidative stress in the progression of pancreatic beta-cell dysfunction in type 2 diabetes, we evaluated the effects of antioxidants in diabetic C57BL/KsJ-db/db mice. According to an intraperitoneal glucose tolerance test, the treatment with antioxidants retained glucose stimulated insulin secretion and moderately decreased blood glucose levels. Histological analyses of the pancreata revealed that the beta-cell mass was significantly larger in the mice treated with the antioxidants, and the antioxidant treatment suppressed apoptosis in beta-cells without changing the rate of beta-cell proliferation. The antioxidant treatment also preserved the amounts of insulin content and insulin mRNA, making the extent of insulin degranulation less evident. As possible mechanism underlying the phenomena, expression of pancreatic and duodenal homeobox factor-1 (also known as IDX-1/STF 1/IPF1), an important transcription factor for the insulin gene, was more clearly visible in the nuclei of islet cells after the antioxidant treatment. Under diabetic conditions, JNK is activated by oxidative stress and involved in the suppression of insulin gene expression. This JNK effect appears to be mediated in part by nucleocytoplasmic translocation of PDX-1, which is also downstream of JNK activation. Taken together, oxidative stress and consequent activation of the JNK pathway are involved in progression of beta-cell dysfunction found in diabetes. Antioxidants may serve as a novel mechanism-based therapy for type 2 diabetes. PMID- 15126295 TI - Initiation of apoptotic signal by the peroxidation of cardiolipin of mitochondria. AB - Overexpression of phospholipid hydroperoxide glutathione peroxidase (PHGPx) in mitochondria of RBL2H3 cells (M15 cells) prevented the release of cytochrome c (cyt.c), the activation of caspase-3, and apoptosis caused by 2-deoxyglucose (2DG), whereas cells overexpressing nonmitochondrial PHGPx(L9) and control (S1) cells were induced to apoptosis. Hydro-peroxide levels in mitochondria of L9 and S1 cells were significantly enhanced by 2DG-induced apoptosis. In contrast, generation of hydroperoxide in mitochondria was protected in M15 cells, which also showed resistance to apoptosis by etoposide, staurosporine, UV irradiation, cycloheximide, and actinomycin D, stimuli that induce apoptosis by the liberation of cyt.c from mitochondria. Cyt.c preferentially binds to the monolayer of cardiolipin (CL), the specific phospholipid of the inner membrane of mitochondria. The amount of cyt.c bound to the monolayer of cardiolipin hydroperoxide (CL-OOH) was much lower than that bound to CL. Cyt.c bound to liposome containing CL was released by peroxidation with a radical initiator. Adenine nucleotide translocator (ANT), which regulates the opening and closing the permeability transition (PT) pore, potentially was inactivated in apoptosis induced S1 cells 4 h after the addition of 2DG, coincidentally with cyt.c release from mitochondria. ANT activity was suppressed by the fusion of isolated mitochondria with liposomes containing CL-OOH. ANT activity was expressed in proteoliposomes containing 10% CL, but it was competitively inhibited by the addition of CL-OOH. This study suggests that CL peroxidation might have an initiating role in the liberation of cyt.c from the inner membrane, and in the opening of the PT pore via inactivation of ANT. Mitochondrial PHGPx might play a role as an anti-apoptotic factor by protecting CL and reducing CL-OOH. PMID- 15126296 TI - Diabetes mellitus with mitochondrial gene mutations in Japan. AB - Diabetes mellitus due to the mitochondrial DNA 3243(A-G) mutation is reported to represent 0.5-1% of the general diabetic population in Japan. To further elucidate the clinical symptoms and course of diabetes mellitus with the 3243 mutation, we undertook a nationwide cross-sectional case-finding study and observational study of a genetically defined subject group. One hundred sixteen Japanese diabetic patients with the mutation were registered and analyzed. The patients had a higher maternal inheritance of diabetes or deafness, short stature, thin habitus, and early middle-aged onset of diabetes or deafness. Eighty-six percent of the patients required insulin therapy because of progressive insulin secretory defect. Although half of the patients had the phenotype of type 1 diabetes or slowly progressive type 1 diabetes, the patients lacked the presence of autoantibodies to glutamic acid decarboxylase. Diabetes in the mothers was characterized by early middle-aged onset, reduction in the insulin secretory capacity, early requirement of insulin therapy, and increases in the daily insulin dose. The heteroplasmic ratio of the 3243 mutation in leukocytes was low. The patients had mitochondria-related complications such as sensorineural deafness, cardiomyopathy, cardiac conductance disorders, encephalomyopathy, macular pattern dystrophy, and mental disorders. The patients also had advanced microvascular complications. Thus, this study has revealed that (1) diabetes mellitus with the 3243 mutation is a subtype of diabetes mellitus with mitochondria-related complications and (2) insulin secretory ability is more severely impaired in the patients whose mothers were also diabetic. PMID- 15126297 TI - Accumulation of somatic mutation in mitochondrial DNA and atherosclerosis in diabetic patients. AB - A point mutation of mitochondrial DNA at nucleotide position 3243 A to G is responsible for the genetic cause of diabetes. Otherwise, this mutation is also reported to occur as a somatic mutation, possibility because of oxidative stress. Because diabetes may cause oxidative stress, we hypothesized that accumulation of the somatic A3243G mutation in mitochondrial DNA may be accelerated by diabetes. DNA was extracted from blood samples of 290 nondiabetic healthy subjects (aged 0 60 years) and from 383 type 2 diabetic patients (aged 18-80 years). Then, the extent of somatic A3243G mutation in total mitochondrial DNA was detected by real time polymerase chain reaction (PCR) using the TaqMan probe. The genotyping of ACE I/D or p22phox C242T was done by PCR or PCR-restriction fragment length polymorphism. Although the level of the A3243G mutation was negligible in the newborn group, it increased in healthy subjects aged 20-29 and 41-60 years. In diabetic patients, the mutational rate increased along with age and the duration of diabetes. In the middle-aged group (41-60 years old), the A3243G mutation accumulates fourfold higher in the diabetic patients than in the healthy subjects. Moreover, multiple regression analysis revealed that the most critical factor associated with this mutation in diabetic patients was the duration of diabetes. Furthermore, the genotype of DD, DI-CC (ACE-p22phox) has the highest mutational rate and the thickest intima-media thickness of the carotid artery. In conclusion, diabetes accelerates the accumulation of the somatic A3243G mutation in mitochondrial DNA, and this somatic mutation may be a marker for the duration of diabetes and atherosclerosis. PMID- 15126299 TI - Mitochondrial encephalomyopathies: diagnostic approach. AB - Mitochondrial diseases have extremely heterogeneous clinical presentations due to the ubiquitous nature of mitochondria and the dual genetic control of the respiratory chain. Thus, mitochondrial disorders can be multisystemic (mitochondrial encephalomyopathies) or confined to a single tissue, and they can be sporadic or transmitted by mendelian or maternal inheritance. Mendelian disorders are usually inherited as autosomal recessive traits, tend to present earlier in life, and usually "breed true" in each family. By contrast, mitochondrial DNA-related diseases usually start later and vary in their presentation within members of the same family. Precise diagnosis is often a challenge; we go through the traditional steps of the diagnostic process, trying to highlight clues to mitochondrial dysfunction in the family history, physical and neurological examinations, routine and special laboratory tests, and histo chemical and biochemical results of the muscle biopsy. The ultimate goal is to reach, whenever possible, a definitive molecular diagnosis, which permits rational genetic counseling and a prenatal diagnosis. PMID- 15126298 TI - Changes of mitochondrial DNA content in the male offspring of protein malnourished rats. AB - Nutritional deprivation of the fetus and infant is associated with susceptibility to the development of impaired glucose tolerance or type 2 diabetes in adult life. Quantitative changes in mitochondrial DNA (mtDNA) seem to be associated with type 2 diabetes, but the effect of protein malnutrition on mtDNA content is not known. This study investigated the effects of protein malnutrition in fetus and early life on mtDNA content and glucose-insulin metabolism in adult life. Male offspring of dams fed a low-protein (LP) diet (8% casein) during pregnancy and lactation were weaned onto either a control (18% casein) diet (recuperated group, R) or a LP diet, and they were compared with the control group (C). The mtDNA content in the liver was lower in the R and LP groups than in the C group at 5 weeks of age, but higher in the R and LP groups than in the C group at 15 weeks of age. The mtDNA content in skeletal muscle and pancreas was significantly lower in the R and LP groups than in the C group at 25 weeks of age. Fetal malnourished rats showed decreased pancreatic beta-cell mass and reduced insulin secretory responses to glucose load, but no differences in glucose tolerance or insulin sensitivity. Our findings imply that protein malnutrition in utero causes changes in mtDNA content, impaired beta-cell development, and insulin secretion, which may contribute to the development of type 2 diabetes in later life. PMID- 15126300 TI - Mitochondrial encephalomyopathies: therapeutic approach. AB - Therapy for mitochondrial diseases is woefully inadequate. How-ever, lack of cure does not equate with lack of treatment. In this review, we consider sequentially several different therapeutic approaches. Palliative therapy is dictated by good medical practice and includes anticonvulsant medication, control of endocrine dysfunction, and surgical procedures. Removal of noxious metabolites is centered on combating lactic acidosis, but it extends to other metabolites, such as thymidine in patients with the mitochondrial neurogastrointestinal encephalomyopathy syndrome. Attempts to bypass blocks in the respiratory chain by administration of artificial electron acceptors have not been successful, but this concept may be amenable to genetic engineering. Administration of metabolites and cofactors is the mainstay of real-life therapy and includes both components of the respiratory chain and other natural compounds. There is increasing interest in the administration of reactive oxygen species scavengers both in primary mitochondrial diseases and in neurodegenerative diseases directly or indirectly related to mitochondrial dysfunction. Aerobic exercise and physical therapy prevent or correct deconditioning and improve exercise tolerance in patients with mitochondrial myopathies due to mtDNA mutations. Gene therapy is a challenge because of polyplasmy and heteroplasmy, but interesting experimental approaches are being pursued and include, for example, decreasing the ratio of mutant to wild-type mitochondrial genomes (gene shifting), converting mutated mtDNA genes into normal nDNA genes (allotropic expression), importing cognate genes from other species, or correcting mtDNA mutations with specific restriction endonucleases. Germline therapy raises ethical problems but is being seriously considered to prevent maternal transmission of mtDNA mutations. Preventive therapy through genetic counseling and prenatal diagnosis is still limited for mtDNA-related disorders but is becoming increasingly important for nDNA-related disorders. PMID- 15126301 TI - Comprehensive molecular diagnosis of mitochondrial disorders: qualitative and quantitative approach. AB - Mitochondrial disorders can be caused by mutations in nuclear or mitochondrial encoded genes. Point mutations and large deletions in mitochondrial DNA (mtDNA) are responsible for a small portion of the molecular defects in the mitochondrial oxidative phosphorylation system. A significant number of molecular defects of respiratory chain disorders are probably due to mutations in nuclear genes. Molecular diagnosis of mitochondrial disorders has been difficult because of broad genetic and clinical heterogeneity. Mutational analysis of common point mutations of mtDNA such as A3243G, A8344G, and T8993G/C is routinely performed. However, many patients who clearly have clinical manifestations and muscle pathology consistent with oxidative phosphorylation deficiency do not have detectable common mtDNA point mutations. A more comprehensive mutation screening method, temporal temperature gradient gel electrophoresis, was used to scan for unknown mutations in the entire mitochondrial genome. Novel mutations have been discovered but only account for a small portion of patients with suspected mitochondrial RC disorders. Real-time quantitative PCR analysis was used to measure cellular mtDNA content. Abnormal levels of mtDNA were found in many patients with respiratory chain disorders. Molecular analysis revealed that mutations in the thymidine phosphorylase gene are not seen in young patients with severe mtDNA depletion who do not demonstrate clinical features of mitochondrial neurogastrointestinal encephalomyopathy. It was also noted that an increase in the size or number of mitochondria was not necessarily associated with an increase in mtDNA content. On the contrary, in some cases the mtDNA was depleted. Respiratory activity in patients with a defective mitochondrial genome due to either point mutations or deletions may be compensated by amplification of mtDNA. Therefore, a comprehensive molecular analysis of mitochondrial respiratory chain disorders should include qualitative identification of the mutation and quantitative measurement of both the degree of mutant heteroplasmy and the total amount of mtDNA. PMID- 15126302 TI - Molecular pathogenetic mechanism of maternally inherited deafness. AB - Mutations in the mitochondrial DNA (mtDNA) have been shown to be one important cause of deafness. In particular, mutations in the mtDNA have been associated with both syndromic and nonsyndromic forms of sensori-neural hearing loss. The deafness-linked mutations often occur in the mitochondrial 12S rRNA gene and in tRNA genes. Mutations in the 12S rRNA gene account for most of the cases of aminoglycoside ototoxicity. The other hot spot for mutations associated with hearing impairment is the tRNA(Ser(UCN)) gene, as five deafness-linked mutations have been identified in this gene. Nonsyndromic deafness-linked mtDNA mutations are often homoplasmic or at high levels of heteroplasmy, indicating a high threshold for pathogenicity. Phenotypic expression of these mtDNA mutations requires the contribution of other factors such as nuclear modifier gene(s), environmental factor(s), or mitochondrial haplotype(s). PMID- 15126303 TI - Genetic and functional analysis of mitochondrial DNA-encoded complex I genes. AB - Mammalian mitochondrial NADH dehydrogenase (complex I) is a multimeric complex consisting of at least 45 subunits, 7 of which are encoded by mitochondrial DNA (mtDNA). The function of these subunits is largely unknown. We have established an efficient method to isolate and characterize cells carrying mutations in various mtDNA-encoded complex I genes. With this method, 15 mouse cell lines with deficiencies in complex I-dependent respiration were obtained, and two near homoplasmic mutations in mouse ND5 and ND6 genes were isolated. Furthermore, by generating a series of cell lines with the same nuclear background but different content of an mtDNA nonsense mutation, we analyzed the genetic and functional thresholds in mouse mitochondria. We found that in wild-type cells, about 40% of ND5 mRNA is in excess of that required to support a normal rate of ND5 subunit synthesis. However, there is no indication of compensatory upsurge in either transcription or translation with the increase in the proportion of mutant ND5 genes. Interestingly, the highest ND5 protein synthesis rate was just sufficient to support the maximum complex I-dependent respiration rate, suggesting a tight regulation at the translational level. In another line of research, we showed that the mitochondrial NADH-quinone oxidoreductase of Saccharomyces cerevisiae (NDI1), although consisting of a single subunit, can completely restore respiratory NADH dehydrogenase activity in mutant human cells that lack the essential mtDNA-encoded subunit ND4. In particular, in these transfected cells, the yeast enzyme becomes integrated into the human respiratory chain and fully restores the capacity of the cells to grow in galactose medium. PMID- 15126304 TI - Genome-wide analysis of signal transducers and regulators of mitochondrial dysfunction in Saccharomyces cerevisiae. AB - Mitochondrial dysfunction is a hallmark of cancer cells. However, genetic response to mitochondrial dysfunction during carcinogenesis is unknown. To elucidate genetic response to mitochondrial dysfunction we used Saccharomyces cerevisiae as a model system. We analyzed genome-wide expression of nuclear genes involved in signal transduction and transcriptional regulation in a wild-type yeast and a yeast strain lacking the mitochondrial genome (rho(0)). Our analysis revealed that the gene encoding cAMP-dependent protein kinase subunit 3 (PKA3) was upregulated. However, the gene encoding cAMP-dependent protein kinase subunit 2 (PKA2) and the VTC1, PTK2, TFS1, CMK1, and CMK2 genes, involved in signal transduction, were downregulated. Among the known transcriptional factors, OPI1, MIG2, INO2, and ROX1 belonged to the upregulated genes, whereas MSN4, MBR1, ZMS1, ZAP1, TFC3, GAT1, ADR1, CAT8, and YAP4 including RFA1 were downregulated. RFA1 regulates DNA repair genes at the transcriptional level. RFA is also involved directly in DNA recombination, DNA replication, and DNA base excision repair. Downregulation of RFA1 in rho(0) cells is consistent with our finding that mitochondrial dysfunction leads to instability of the nuclear genome. Together, our data suggest that gene(s) involved in mitochondria-to-nucleus communication play a role in mutagenesis and may be implicated in carcinogenesis. PMID- 15126305 TI - Enhanced detection of deleterious mutations by TTGE analysis of mother and child's DNA side by side. AB - Mitochondrial DNA (mtDNA) disorders represent a group of heterogeneous diseases that are caused by mutations in mtDNA. We examined 45 pairs of mother and the affected child, by screening the entire mitochondrial genome with temporal temperature gradient gel electrophoresis (TTGE), using 32 pairs of overlapping primers. TTGE is an effective method of mutation detection. It detects and distinguishes heteroplasmic mutations from homoplasmic mutations. By running the mother and child's DNA samples side by side and sequencing only the DNA fragments showing different TTGE patterns, excessive sequencing can be avoided, particularly because most sequence variations represent benign polymorphisms. Mutations identified by sequencing were further confirmed by PCR/ASO (allele specific oligonucleotide) dot blot analysis or PCR/RFLP (restriction fragment length polymorphism). A total of seven differences in sequence between mother and child pairs were identified: A189G, T5580C, G5821A, C5840T, A8326G, G12207A, and G15995A. All but two mutations were novel. The most significant are the A8326G, G12207A, and G15595A mutations. The A8326G is located at the anticodon region of tRNA(Lys), right next to the first nucleotide of the triplet codon, and it is highly conserved throughout evolution. The G12207A mutation is located at the first base of tRNA(ser) (AGY). The G15995A mutation occurs at a stem region that results in the disruption of the first base pair at the anticodon loop of tRNA(Pro) and is highly conserved throughout evolution from sea urchins to mammals. Running TTGE side by side with DNAs from mother and the affected child is a novel method to detect deleterious mutations. PMID- 15126306 TI - Quantitative PCR analysis of mitochondrial DNA content in patients with mitochondrial disease. AB - Molecular diagnosis of mitochondrial DNA disorder is usually focused on point mutations and large deletions. In the absence of detectable mtDNA mutations, abnormal amounts of mtDNA, either depletion or elevation, can be indicative of mitochondrial dysfunction. The amount of mitochondrial DNA (mtDNA), however, varies among individuals of different ages and among different tissues within the same individual. To establish a range of mtDNA levels, we analyzed 300 muscle and 200 blood specimens from patients suspected of having a mitochondrial disorder by real-time quantitative polymerase chain reaction (PCR) method. Copy numbers were calculated from the standard curve and threshold cycle number using TaqMan probes; 6FAM 5'TTACCGGGCTCTGCCATCT3'-TAMRA and VIC-5'AGCAATAACAGGTCTGTGATG3' TAMRA for mtDNA and 18S rRNA gene (nDNA), respectively. The copy number ratio of mtDNA to nDNA was used as a measure of mtDNA content in each specimen. The mtDNA content in muscle increases steadily from birth to about 5 years of age; thereafter, it stays about the same. On the contrary, the mtDNA content in blood decreases with age. The amount of mtDNA in skeletal muscle is about 5-20 times higher than that in blood. About 7% of patients had mtDNA levels in muscle below 20% of the mean of the age-matched group, and about 10% of patients had muscle mtDNA levels 2- to 16-fold higher than the mean of the age-matched group. Patients with abnormal levels of mtDNA, either depletion or proliferation, had significant clinical manifestations characteristic of mitochondrial disease in addition to abnormal respiratory enzymes and mitochondrial cytopathies. Cardiomyopathy, lactic acidosis, abnormal brain MRI findings, hypotonia, developmental delay, seizures, and failure to thrive are general clinical pictures of patients with mtDNA depletion. The average age of patients with mtDNA depletion is 4.1 years, compared to 23.6 years in patients with mtDNA proliferation. Mutations in nuclear genes involved in mtDNA synthesis and deoxynucleotide pools are probably the cause of mtDNA depletion. Our results demonstrate that real time quantitative PCR is a valuable tool for molecular screening of mitochondrial diseases. PMID- 15126307 TI - Somatic mitochondrial DNA mutations in oral cancer of betel quid chewers. AB - Somatic mitochondrial DNA alteration is a general phenomenon that occurs in cancerous cells. Although numerous mtDNA mutations have been identified in various tumors, the pathogenic significance of these mutations remains unclear. In order to better understand the role of mtDNA mutations in the neoplastic process of oral cancer, the occurrence of mtDNA mutations in oral squamous cell carcinomas was screened by temporal temperature gradient gel electrophoresis (TTGE). The entire mitochondrial genome was amplified with 32 pairs of overlapping primers. The DNA fragments showing different banding patterns between normal and tumor mtDNA were sequenced for the identification of the mutations. Fourteen of 18 (77.8%) tumors had somatic mtDNA mutations with a total of 26 mutations. Among them, 6 were in mRNA coding region. Three were missense mutations (C14F, H186R, T173P) in NADH dehydrogenase subunit 2 (ND2). One frameshift mutation, 9485delC, was in cytochrome c oxidase subunit III. Eight (44%) tumors had insertion or deletion mutations in the np303-309 poly C region of the D-loop. Our results demonstrate that somatic mtDNA mutations occur in oral cancer. The missense and frameshift mutations in the evolutionary conserved regions of the mitochondrial genome may have functional significance in the pathogenesis of oral cancer. PMID- 15126308 TI - Association of the mitochondrial DNA 16189 T to C variant with lacunar cerebral infarction: evidence from a hospital-based case-control study. AB - A transition of T to C at nucleotide position 16189 in the hypervariable D-loop region of mitochondrial DNA (mtDNA) has attracted research interest for its probable correlation with increasing insulin resistance and development of diabetes mellitus (DM) in adult life. In this article, we present our observations of the positive relationship between this variant and cerebral infarction. Six hundred and one subjects in two groups-one with cerebral infarction (307 cases), the other with no cerebral infarction (294 cases)-were recruited. Their clinical features, fasting blood sugar and insulin levels, and insulin resistance index, were recorded. Patients with cerebral infarction were further categorized into four different subgroups according to the TOAST criteria for stroke classification. The results showed the occurrence of the mtDNA 16189 variant in 34.2% of patients with cerebral infarction and in 26.5% of normal controls. The difference in the occurrence rates between the two groups was statistically significant (P = 0.041). Further studies of the occurrence rate in each stroke subgroup revealed that the variant occurred at the highest frequency in the small vessel subgroup (41.5%). The difference in occurrence rate between this subgroup and the normal controls is highly significant (P = 0.006). These results correlated well with the findings of significantly increased levels of average fasting blood insulin and a higher index of average insulin resistance in the small vessel subgroup of patients harboring this mtDNA variant. Taken together, we suggest that the mtDNA 16189 variant is a predisposing genetic factor for the development of insulin resistance and may be related to various phenotypic expressions in adult life such as development of DM and vascular pathologies involved in stroke and cardiovascular diseases. PMID- 15126309 TI - Mechanisms of cell death induced by cadmium and arsenic. AB - Cadmium (Cd) and arsenic (As) are known toxic metals in humans. As trioxide (As(2)0(3)) has been recently used as a mitochondria-targeting drug in acute promyelocytic leukemia. In the present study, we examined the intracellular action of these metals using rat kidney tubular cells and cells tolerant to the metals. The cells were cultured with CdCl(2) (1-10 micro M) or As(2)O(3) (1-2.5 micro M). Cells tolerant to Cd and As (Cd-T and As-T, respectively) were defined as cells that survived at toxic concentrations of each metal. Both Cd and As induced cell toxicity in a dose-dependent fashion, which was accompanied by fragmented DNA and decreased mitochondrial membrane potential. Intracellular glutathione (GSH) increased with the increase of Cd and As concentration. In Cd-T and As-T cells, GSH levels were twice those observed in normal cells. When each metal-tolerant culture was exposed to the other different metal, i.e., As or Cd, the protective property was maintained. However, when buthionine sulfoximine (BSO) was added to the metal-tolerant cultures, apoptosis was restored in both Cd T and As-T. Our results indicate that (1) although GSH is increased in NRK52E by the addition of Cd and As, mitochondria-mediated apoptosis can be still induced, (2) the protective property against metal-induced cytotoxicity is identical in Cd T and As-T cultures, and (3) although GSH was higher in the metal-tolerant cell lines, depression of GSH by BSO induced apoptosis. We conclude that Cd- and As induced apoptosis is mediated by an identical mechanism involving intracellular GSH reactive oxidation. PMID- 15126310 TI - Cadmium-induced nephropathy in rats is mediated by expression of senescence associated beta-galactosidase and accumulation of mitochondrial DNA deletion. AB - Long-term exposure to cadmium (Cd) induces perturbation of kidney proximal tubular epithelial cells. Mitochondrial dysfunction in renal cortical cells may contribute to the pathogenesis of Cd-induced nephropathy. In this study, we examined the accumulation of mitochondrial DNA (mtDNA) with a large deletion and cellular senescence in the renal cortex. Wistar rats at 8 weeks of age were intraperitoneally injected with 1 mL of 1 mM CdCl(2) or saline, 3 times/week for 5, 20, 40, or 80 weeks. Mitochondrial Cd content in the renal cortex was quantified by atomic absorption analysis. Cytochrome c oxidase (CCO) and senescence-associated beta-galactosidase (SA-beta-gal) activity were determined in renal cortex by enzyme-histochemistry. mtDNA in total DNA extracted from the renal cortex was amplified by PCR, and mtDNA deletions, including 4,834-bp (nt8118-nt12937) deletion, were determined and semiquantified. After 40 weeks of Cd injection, Cd levels in the renal cortex reached a saturation level, and 30% of the level of the whole-cell fraction was found in the mitochondria. CCO activity in the renal cortex, which was predominantly found in proximal tubular cells, decreased after 40 weeks of Cd exposure. Expression of SA-beta-gal was detected primarily in the proximal tubular cells and significantly increased after 80 weeks of Cd exposure. After 40 weeks of study, accumulation of 4,834-bp deletion in mtDNA was evident in both groups of rats; however, the amount of the deletion was significantly greater in Cd-treated rats than in control rats. Our results indicate that long-term Cd exposure induced a post-regenerative state of proximal tubular cells, which accelerated accumulation of 4,834-bp mtDNA deletions in the renal cortex, suggesting that Cd may be a senescence acceleration factor for kidney proximal tubular epithelial cells, which results in Cd-induced nephropathy. PMID- 15126311 TI - Investigation of common mitochondrial point mutations in Korea. AB - Between 1997 and 2002, 65 patients with suspected mitochondrial diseases were screened for the mitochondrial point mutations A3243G, T3271C, A8344G, and T8356C. Among these patients, 15 were found to have one of these mutations: 12 with A3243G and 3 with A8344G. The phenotypes of A3243G and A8344G mutations were MELAS and MERRF, respectively. Many asymptomatic family members had the same mutations. In this report, detailed clinical and laboratory findings are presented. PMID- 15126312 TI - Leber's hereditary optic neuropathy: the spectrum of mitochondrial DNA mutations in Iranian patients. AB - We studied 14 patients with Leber's hereditary optic neuropathy (LHON) to investigate the mtDNA haplotypes associated with the primary mutation(s). Eleven patients carried the mitochondrial DNA (mtDNA) G11778A mutation, while one had the T14484C mutation; one patient had the G3460A mutation and one the G14459A mutation. The Iranian G11778A LHON mutation was not associated with two mtDNA haplogroups-M (0.0% compared with 3.2% in healthy controls) and J (7.7% compared with 10% in healthy controls). Our results showed a similarity in the pattern of LHON primary point mutations between Iranian families with LHON and those of Russian, European, and North American origin. Our results also do not support an association between mtDNA haplogroups J and M with LHON primary point mutations. PMID- 15126313 TI - Randomised controlled trial of effects of Helicobacter pylori infection and its eradication on heartburn and gastro-oesophageal reflux: Bristol helicobacter project. AB - OBJECTIVES: To investigate the effects of Helicobacter pylori infection and its eradication on heartburn and gastro-oesophageal reflux. DESIGN: Cross sectional study, followed by a randomised placebo controlled trial. SETTING: Seven general practices in Bristol, England. PARTICIPANTS: 10,537 people, aged 20-59 years, with and without H pylori infection (determined by the (13)C-urea breath test). MAIN OUTCOME MEASURES: Prevalence of heartburn and gastro-oesophageal acid reflux at baseline and two years after treatment to eradicate H pylori infection. RESULTS: At baseline, H pylori infection was associated with increased prevalence of heartburn (odds ratio 1.14, 95% confidence interval 1.05 to 1.23) but not reflux (1.05, 0.97 to 1.14). In participants with H pylori infection, active treatment had no effect on the overall prevalence of heartburn (0.99, 0.88 to 1.12) or reflux (1.04, 0.91 to 1.19) and did not improve pre-existing symptoms of heartburn or reflux. CONCLUSIONS: H pylori infection is associated with a slightly increased prevalence of heartburn but not reflux. Treatment to eradicate H pylori has no net benefit in patients with heartburn or gastro-oesophageal reflux. PMID- 15126314 TI - Chimerism and clinical outcomes of 110 recipients of unrelated donor bone marrow transplants who underwent conditioning with low-dose, single-exposure total body irradiation and cyclophosphamide. AB - We hypothesized that low-dose (550-cGy), single-exposure, high dose rate (30 cGy/min) total body irradiation (TBI) with cyclophosphamide as conditioning for HLA-compatible unrelated donor (URD) bone marrow transplantation (BMT) would result in donor chimerism (DC) with a low risk for serious organ toxicity and treatment-related mortality (TRM). Twenty-six patients with good risk diagnoses (acute leukemia in first complete remission [CR] and chronic-phase chronic myelogenous leukemia [CML]) and 84 with poor risk diagnoses underwent this regimen and URD BMT. Unsorted marrow nucleated cells were assessed for chimerism using VNTR probes. All DC occurred in 78 (86%) of 91 evaluable patients at 1 or more follow-up points. Graft failure occurred in 7 (7.7%) patients. Fatal organ toxicity occurred in only 2% of patients. TRM rates through 2 years of follow-up were 19% and 42% in those with good and poor risk diagnoses, respectively. Overall and disease-free survival rates in the good risk group were 47% and 40%, respectively, and in the poor risk group they were 25% and 21%, respectively, at a median follow-up for living patients of 850 days (range, 354-1588 days). This regimen resulted in 100% DC in most patients undergoing URD BMT with a relatively low risk for fatal organ toxicity and TRM. PMID- 15126315 TI - Expression of the chemokine receptor CCR2 on immature B cells negatively regulates their cytoskeletal rearrangement and migration. AB - Immature B cells are targeted to specific areas in the spleen, where a fraction of these cells receive signals that induce them to mature and participate in the immune response. In this study, we show that the C-C chemokine receptor 2 (CCR2) is transcribed in immature B cells, while its message is dramatically down regulated at the mature stage. CCR2-deficient cells exhibit up-regulation of chemokine-induced actin polymerization, migration, and homing to the lymph nodes of immature B cells. In addition, we demonstrate that control of homing by CCR2 is mediated by its ligand, CCL2/JE, which is secreted by B cells and down regulates the stromal derived factor-1 (SDF-1) signaling cascade. Thus, this study describes an additional, previously uncharacterized, role for CCR2 and its ligand as negative regulators of the homing of immature B cells. PMID- 15126316 TI - Rituximab antiproliferative effect in B-lymphoma cells is associated with acid sphingomyelinase activation in raft microdomains. AB - Rituximab is a chimeric human immunoglobulin G1 (IgG1) anti-CD20 monoclonal antibody with significant activity against CD20+ malignant B cells. Rituximab is currently used with success in the treatment of B-cell-derived lymphoid neoplasias either alone or in combination with chemotherapy. However, the predominant mechanism by which rituximab exerts its antitumor properties in vivo remains unknown. In the present study, we demonstrate that in Daudi and RL B lymphoma cells, rituximab (without cross-linking) used at the saturating dose of 10 microg/mL induced moderate accumulation in G1 phase, growth inhibition, and significant loss in clonogenic potential. However, in these cells, rituximab induced no apoptosis. Furthermore, we observed that treatment with rituximab resulted in a rapid and transient increase in acid-sphingomyelinase (A-SMase) activity and concomitant cellular ceramide (CER) generation in raft microdomains. We also observed that rituximab-treated cells externalized both A-SMase and CER that colocalized with the CD20 receptor. Finally, we present evidence that rituximab-induced growth inhibition may be mediated through a CER-triggered signaling pathway, leading to the induction of cell cycle-dependent kinase inhibitors such as p27Kip1 through a mitogen-activated protein kinase (MAPK) dependent mechanism. PMID- 15126317 TI - In vitro studies of a FLT3 inhibitor combined with chemotherapy: sequence of administration is important to achieve synergistic cytotoxic effects. AB - Patients with acute myeloid leukemia (AML) harboring internal tandem duplication mutations of the FLT3 receptor (FLT3/ITD mutations) have a poor prognosis compared to patients lacking such mutations. Incorporation of FLT3 inhibitors into existing chemotherapeutic regimens has the potential to improve clinical outcomes in this high-risk group of patients. CEP-701, an indolocarbazole-derived selective FLT3 inhibitor, potently induces apoptosis in FLT3/ITD-expressing cell lines and primary leukemic blasts. We conducted a series of in vitro cytotoxicity experiments combining CEP-701 with chemotherapy using the FLT3/ITD-expressing cell lines MV4-11 and BaF3/ITD as well as a primary blast sample from a patient with AML harboring a FLT3/ITD mutation. CEP-701 induced cytotoxicity in a synergistic fashion with cytarabine, daunorubicin, mitoxantrone, or etoposide if used simultaneously or immediately following exposure to the chemotherapeutic agent. In contrast, the combination of pretreatment with CEP-701 followed by chemotherapy was generally antagonistic, particularly with the more cell cycle dependent agents such as cytarabine. This effect appears to be due to CEP-701 causing cell cycle arrest. We conclude that in FLT3/ITD-expressing leukemia cells, CEP-701 is synergistic with standard AML chemotherapeutic agents, but only if used simultaneously with or immediately following the chemotherapy. These results should be considered when designing trials combining chemotherapy with each of the FLT3 inhibitors currently in clinical development. PMID- 15126318 TI - Identification of novel mutations in ADAMTS13 in an adult patient with congenital thrombotic thrombocytopenic purpura. AB - Congenital thrombotic thrombocytopenic purpura/hemolytic uremic syndrome (TTP/HUS) is associated with an inherited von Willebrand factor-cleaving protease (ADAMTS13 [a disintegrin and metalloprotease with thrombospondin type I domains 13]) deficiency. In this study, we identified novel mutations in the ADAMTS13 gene in a patient with TTP. The patient was a 51-year-old Japanese male who exhibited TTP symptoms at frequent intervals. The ADAMTS13 activity during acute episodes was less than 3% that of normal. The enzyme activities of the patient's father and mother were both 46%, and both parents were asymptomatic. Genetic analysis revealed that the patient was a compound heterozygote for 2 mutations. One mutation was a missense mutation in the metalloprotease domain (A250V, exon 7), and the other was a guanine to adenine substitution at the 5' end of intron 3 (intron 3 G-->A). In vitro expression studies revealed that the A250V mutation markedly reduced ADAMTS13 activity and the intron 3 G-->A mutation caused abnormal mRNA synthesis. PMID- 15126319 TI - PECAM-1 is expressed on hematopoietic stem cells throughout ontogeny and identifies a population of erythroid progenitors. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1) (CD31) is an adhesion molecule expressed on endothelial cells and subsets of leukocytes. Analysis of phenotypically defined hematopoietic stem cells (HSCs) from the yolk sac, fetal liver, and adult bone marrow demonstrates CD31 expression on these cells throughout development. CD31+ c-kit+ cells, but not CD31- c-kit+ cells, isolated from day-9.5 yolk sac give rise to multilineage hematopoiesis in vivo. Further evaluation of the CD31+ lineage marker-negative fraction of adult bone marrow reveals functionally distinct cell subsets. Transplantation of CD31+ Lin- c-kit- cells fails to protect lethally irradiated recipients, while CD31+ Lin- c-kit+ Sca-1- cells (CD31+ Sca-1-) provide radioprotection in the absence of long-term donor-derived hematopoiesis. Although donor-derived leukocytes were not detected in CD31+ Sca-1- recipients, donor-derived erythroid cells were transiently produced during the initial phases of bone marrow recovery. These results demonstrate CD31 expression on hematopoietic stem cells throughout ontogeny and identify a population of CD31+ short-term erythroid progenitors cells that confer protection from lethal doses of radiation. PMID- 15126320 TI - The impact of low-dose busulfan on clonal dynamics in nonhuman primates. AB - An understanding of the number and contribution of individual pluripotent hematopoietic stem cells (HSCs) to the formation of blood lineages has important clinical implications for gene therapy and stem cell transplantation. We have been able to efficiently mark rhesus macaque long-term repopulating stem and progenitor cells with retroviral vectors, and track their in vivo contributions to hematopoiesis using the linear amplification mediated-polymerase chain reaction (LAM-PCR) technique of insertion site analysis. We assessed the impact of busulfan on contributions of individual retrovirally marked clones to hematopoiesis. There were 2 macaques that received transplants of retrovirally transduced CD34(+) cells 2 years previously that were then treated with 4 mg/kg busulfan. Despite only transient and mild suppression of peripheral blood counts, the numbers of individual stem/progenitor clones contributing to granulocyte production decreased dramatically, by 80% in the first monkey and by 60% in the second monkey. A similar impact was seen on clones contributing to T cells. The clone numbers recovered gradually back toward baseline by 5 months following busulfan in the first monkey and by 3 months in the second monkey, and have remained stable for more than one year in both animals. Tracking of individual clones with insertion-site-specific primers suggested that clones contributing to hematopoiesis prior to busulfan accounted for the majority of this recovery, but that some previously undetected clones began to contribute during this recovery phase. These results indicate that even low-dose busulfan significantly affects stem and progenitor cell dynamics. The clonal diversity of hematopoiesis was significantly decreased after even a single, clinically well-tolerated dose of busulfan, with slow but almost complete recovery over the next several months, suggesting that true long-term repopulating stem cells were not permanently deleted. However, the prolonged period of suppression of many clones suggests that transplanted HSCs may have a marked competitive advantage if they can engraft and proliferate during this time period, and supports the use of this agent in nonmyeloablative regimens PMID- 15126321 TI - Syndecan-1 up-regulated by ephrinB2/EphB4 plays dual roles in inflammatory angiogenesis. AB - EphrinB2 and EphB4, its cognate receptor, are important in the vascular development of the mouse embryo. Their roles in human inflammatory angiogenesis, however, are not well understood. By examining hyperinflammatory lesions, we saw that ephrinB2 was predominantly expressed in macrophage-like cells and EphB4 in small venules. Because macrophages usually transmigrate through postcapillary venules during inflammation, we wanted to explore the downstream effects of EphB4 after binding to ephrinB2. By using cDNA microarray technique and following reverse transcriptase-polymerase chain reaction (RT-PCR), we found that syntenin and syndecan-1 were up-regulated in EphB4-positive endothelial cells dose dependently and time dependently after stimulation with preclustered ephrinB2. In vitro, ephrinB2 suppressed the angiogenic effects of basic fibroblast growth factor (bFGF) on EphB4-positive endothelial cells, partially due to syndecan-1's competition with fibroblast growth factor receptor (FGFR) for bFGF. However, ephrinB2 exhibited angiogenic effects in vivo, possibly due to an inflammation associated enzyme-heparanase. The enzymes could convert the inhibitory effect of ephrinB2 on EphB4-positive endothelial cells to an activating effect by removing poorly sulfated side chains of up-regulated syndecan-1 ectodomain. Depending on the presence of heparanases, the roles of syndecan-1 may be opposite in different physiological settings. PMID- 15126322 TI - Inhibition of intravascular thrombosis in murine endotoxemia by targeted expression of hirudin and tissue factor pathway inhibitor analogs to activated endothelium. AB - We have generated transgenic mice expressing the leech anticoagulant hirudin and human tissue factor pathway inhibitor tethered to the cell surface by fusion with fragments of human CD4 and P-selectin. Expression of the transgenes is under the control of the CD31 (platelet endothelial cell adhesion molecule [PECAM]) promoter, limiting expression to endothelial cells, monocytes, and platelets. In addition, the P-selectin sequence directs expression to secretory granules. Functional cell surface expression only occurs when the cells are activated. In a mouse model of systemic lipopolysaccharide (LPS)-induced endotoxemia, we show that expression of either anticoagulant on activated endothelium inhibits the widespread intravascular thrombosis, thrombocytopenia, and consumptive coagulopathy associated with endotoxemia. Importantly, non- LPS-treated transgenic mice had normal baseline bleeding times. We speculate that targeted delivery of anticoagulants to the endothelium may be a strategy worth pursuing in clinical sepsis to improve efficacy of systemic anticoagulation while minimizing potential hemorrhagic side effects. PMID- 15126323 TI - Follicular lymphoma international prognostic index. AB - The prognosis of follicular lymphomas (FL) is heterogeneous and numerous treatments may be proposed. A validated prognostic index (PI) would help in evaluating and choosing these treatments. Characteristics at diagnosis were collected from 4167 patients with FL diagnosed between 1985 and 1992. Univariate and multivariate analyses were used to propose a PI. This index was then tested on 919 patients. Five adverse prognostic factors were selected: age (> 60 years vs < or = 60 years), Ann Arbor stage (III-IV vs I-II), hemoglobin level (< 120 g/L vs > or = 120 g/L), number of nodal areas (> 4 vs < or = 4), and serum LDH level (above normal vs normal or below). Three risk groups were defined: low risk (0-1 adverse factor, 36% of patients), intermediate risk (2 factors, 37% of patients, hazard ratio [HR] of 2.3), and poor risk (> or = 3 adverse factors, 27% of patients, HR = 4.3). This Follicular Lymphoma International Prognostic Index (FLIPI) appeared more discriminant than the International Prognostic Index proposed for aggressive non-Hodgkin lymphomas. Results were very similar in the confirmation group. The FLIPI may be used for improving treatment choices, comparing clinical trials, and designing studies to evaluate new treatments. PMID- 15126324 TI - Microvessel density as a prognostic factor in women with breast cancer: a systematic review of the literature and meta-analysis. AB - We performed a meta-analysis of all 87 published studies linking intratumoral microvessel density (MVD), reflecting angiogenesis, to relapse-free survival (RFS) and overall survival (OS). With median MVD as cutoff, MVD impact was measured by risk ratio (RR) between the two survival distributions. Seventeen studies did not mention survival data or fit inclusion criteria. Twenty-two were multiple publications of the same series, leaving 43 independent studies (8936 patients). MVD was assessed by immunohistochemistry, using antibodies against factor VIII (27 studies; n = 5262), CD31 (10 studies; n = 2296), or CD34 (8 studies; n = 1726). MVD might be a better prognostic factor when assessed by CD31 or CD34 versus factor VIII (P = 0.11). For RFS, statistical calculations were performed in 25 studies (6501 patients). High MVD significantly predicted poor survival [RR = 1.54 for RFS and OS with the same 95% confidence interval (CI), 1.29-1.84]. Twenty-two studies analyzed separately lymph node-negative patients (n = 3580), for whom predictors of poor survival are requested. This latter meta analysis included 15 studies for RFS (2727 patients) and 11 for OS (1926 patients). High MVD significantly predicted poor survival [RR = 1.99 for RFS (95% CI, 1.33-2.98) and RR = 1.54 for OS (95% CI, 1.01-2.33)]. Between-study variations could result from patient selection criteria, techniques to stain and count microvessels, and cutoff selection. MVD was a significant although weak prognostic factor in women with breast cancer. Standardization of MVD assessment is needed. PMID- 15126325 TI - Tumor suppressor maspin is up-regulated during keratinocyte senescence, exerting a paracrine antiangiogenic activity. AB - Cell senescence is a physiological program of terminal growth arrest, which is believed to play an important role in cancer prevention. Senescent cells secrete multiple growth-regulatory proteins, some of which can affect tumor growth, survival, invasion, or angiogenesis. Changes in expression of different senescence-associated genes were analyzed in cultured human skin keratinocytes (KCs) that underwent replicative senescence or confluence-induced accelerated senescence. Senescent KC cultures showed a strong increase in mRNA and protein expression of maspin, a member of serine protease inhibitor family and an epithelial cell tumor suppressor with anti-invasive and antiangiogenic activities. Immunohistochemical analysis of 14 normal human skin samples (age range from 3 months to 84 years) showed that maspin is expressed by KCs in vivo and that the extent and intensity of maspin expression in the skin is significantly (P = 0.01) correlated with chronological age. Antiangiogenic activity of maspin secreted by senescent KCs was investigated in vitro by testing the effect of conditioned media from different KC cultures on endothelial cell migration in the presence or absence of several angiogenic factors. Media conditioned by senescent cultures (undergoing replicative or accelerated senescence), but not by proliferating KCs, strongly inhibited the stimulation of endothelial cell migration by all of the tested angiogenic factors. Neutralizing antibody against maspin abrogated this effect of conditioned media. These findings indicate that senescent KCs exert a paracrine antiangiogenic activity, and maspin is the principal contributor to this potentially tumor-suppressive effect of cellular senescence. PMID- 15126326 TI - A molecular signature of the Nottingham prognostic index in breast cancer. AB - The Nottingham prognostic index (NPI) is a widely used clinicopathological staging system for breast cancer prognostication. Using a step-wise classification approach where breast tumor expression profiles were first divided into general "molecular subtypes" [estrogen receptor (ER)+, ER-, ERBB2+], followed by an independent analysis of each subtype, we identified a 62-gene expression signature (NPI-ES) highly correlated to the NPI in ER+ tumors. The NPI ES classified the majority of ER+ tumors into two distinct groups with high confidence and was significantly correlated to NPI status in two independent sets of ER+ tumors derived from different centers. The NPI-ES is comparable to the classical NPI in identifying patients likely to exhibit a poor clinical prognosis, as well as to a recently described "prognosis signature" for breast cancer. Our findings demonstrate how expression profiling can complement classical staging systems employing histopathological parameters scored over a continuous range of values. PMID- 15126327 TI - Myristoylation of the fus1 protein is required for tumor suppression in human lung cancer cells. AB - FUS1 is a novel tumor suppressor gene identified in the human chromosome 3p21.3 region that is deleted in many cancers. Using surface-enhanced laser desorption/ionization mass spectrometric analysis on an anti-Fus1-antibody capture ProteinChip array, we identified wild-type Fus1 as an N-myristoylated protein. N-myristoylation is a protein modification process in which a 14-carbon myristoyl group is cotranslationally and covalently added to the NH2-terminal glycine residue of the nascent polypeptide. Loss of expression or a defect of myristoylation of the Fus1 protein was observed in human primary lung cancer and cancer cell lines. A myristoylation-deficient mutant of the Fus1 protein abrogated its ability to inhibit tumor cell-induced clonogenicity in vitro, to induce apoptosis in lung tumor cells, and to suppress the growth of tumor xenografts and lung metastases in vivo and rendered it susceptible to rapid proteasome-dependent degradation. Our results show that myristoylation is required for Fus1-mediated tumor-suppressing activity and suggest a novel mechanism for the inactivation of tumor suppressors in lung cancer and a role for deficient posttranslational modification in tumor suppressor-gene-mediated carcinogenesis. PMID- 15126328 TI - MAT1-modulated cyclin-dependent kinase-activating kinase activity cross-regulates neuroblastoma cell G1 arrest and neurite outgrowth. AB - Cyclin-dependent kinase-activating kinase (CAK) regulates cell cycle G1 exit, where cells commonly commit either to proliferate or to differentiate. CAK activity in G1 regulation is determined by its assembly factor and targeting subunit, menage a trois 1 (MAT1). The precise mechanism of how proliferation/differentiation transition is induced from cancer cell G1 arrest remains unknown. We present evidence that in neuroblastoma CHP126 cells, CAK interacts with and phosphorylates retinoblastoma tumor suppressor protein (pRb) and retinoid X receptor alpha (RXRalpha). Retinoic acid (RA)-induced neuroblastoma cell proliferation/differentiation transition is associated with decreased CAK activity, as evidenced by a switch from CAK hyperphosphorylation of pRb and RXRalpha to hypophosphorylation of pRb and RXRalpha. Manipulation of MAT1 abundance shows that MAT1 reduction mimics RA-induced hypophosphorylation of pRb/RXRalpha, proliferation inhibition, and neurite outgrowth, whereas MAT1 overexpression resists these RA actions. Thus, these findings reveal an important mechanism by which MAT1-modulated CAK activity is crucial in the switch from proliferation to differentiation in neuroblastoma cells. PMID- 15126329 TI - Depletion of methionine aminopeptidase 2 does not alter cell response to fumagillin or bengamides. AB - Inhibition of endothelial cell growth by fumagillin has been assumed to be mediated by inhibition of the molecular target methionine aminopeptidase 2 (MetAp2). New data show that depletion of MetAp2 by siRNA does not inhibit endothelial cell growth. Moreover, MetAp2-depleted endothelial cells remain responsive to inhibition by either fumagillin or a newly identified MetAp2 enzyme inhibitor. These data suggest that MetAp2 function is not required for endothelial cell proliferation. PMID- 15126330 TI - Melanoma differentiation associated gene-7/interleukin-24 promotes tumor cell specific apoptosis through both secretory and nonsecretory pathways. AB - Melanoma differentiation associated gene-7/interleukin-24 (Mda-7/IL-24), a novel member of the IL-10 family of cytokines, uniquely displays cancer-specific apoptosis-inducing activity. Positive results in ongoing phase I/II clinical trials have strengthened the possibility of its utilization as a cancer gene therapeutic. Previous studies document that signaling events leading to Ad.mda-7 induced transformed cell apoptosis are tyrosine kinase-independent. These results suggest that mda-7/IL-24 cancer cell-specific activity could occur through mechanisms independent of binding to its currently recognized cognate receptors and might even occur independent of receptor function. An adenovirus vector expressing a nonsecreted version of MDA-7/IL-24 protein was generated via deletion of its signal peptide. This nonsecreted protein was as effective as wild type secreted MDA-7/IL-24 in inducing apoptosis in prostate carcinoma cell lines and displayed transformed cell specificity and localization of MDA-7/IL-24 in the Golgi/endoplasmic reticulum compartments. Our results indicate that mda-7/IL-24 mediated apoptosis can be triggered through a combination of intracellular as well as secretory mechanisms and can occur efficiently in the absence of protein secretion. PMID- 15126331 TI - Promoter hypermethylation of FANCF: disruption of Fanconi Anemia-BRCA pathway in cervical cancer. AB - Patients with advanced stage invasive cervical cancer (CC) exhibit highly complex genomic alterations and respond poorly to conventional treatment protocols. In our efforts to understand the molecular genetic basis of CC, we examined the role of Fanconi Anemia (FA)-BRCA pathway. Here, we show that FANCF gene is disrupted by either promoter hypermethylation and/or deregulated gene expression in a majority of CC. Inhibition of DNA methylation and histone deacetylases induces FANCF gene re-expression in CC cell lines. FANCF-deregulated CC cell lines also exhibit a chromosomal hypersensitivity phenotype after exposure to an alkylating agent, a characteristic of FA patients. We also show the involvement of BRCA1 gene by promoter hypermethylation or down-regulated expression in a small subset of CC patients. Thus, we have found inactivation of genes in the FA-BRCA pathway by epigenetic alterations in a high proportion of CC patients, suggesting a major role for this pathway in the development of cervical cancer. Thus, these results have important implications in understanding the molecular basis of CC tumorigenesis and clinical management in designing targeted experimental therapeutic protocols. PMID- 15126332 TI - Three classes of genes mutated in colorectal cancers with chromosomal instability. AB - Although most colorectal cancers are chromosomally unstable, the basis for this instability has not been defined. To determine whether genes shown to cause chromosomal instability in model systems were mutated in colorectal cancers, we identified their human homologues and determined their sequence in a panel of colorectal cancers. We found 19 somatic mutations in five genes representing three distinct instability pathways. Seven mutations were found in MRE11, whose product is involved in double-strand break repair. Four mutations were found among hZw10, hZwilch/FLJ10036, and hRod/KNTC, whose products bind to one another in a complex that localizes to kinetochores and controls chromosome segregation. Eight mutations were found in Ding, a previously uncharacterized gene with sequence similarity to the Saccharomyces cerevisiae Pds1, whose product is essential for proper chromosome disjunction. This analysis buttresses the evidence that chromosomal instability has a genetic basis and provides clues to the mechanistic basis of instability in cancers. PMID- 15126333 TI - A hot spot for RAD51C interactions revealed by a peptide that sensitizes cells to cisplatin. AB - DNA repair via the homologous recombination pathway requires the recombinase RAD51 and, in vertabrates, five RAD51 paralogs. The paralogs form two complexes in solution, a XRCC3/RAD51C heterodimer and a RAD51B/RAD51C/RAD51D/XRCC2 heterotetramer. Mutation of any one of the five paralog genes prevents subnuclear assembly of recombinase at damaged sites and renders cells 30-100 fold sensitive to DNA cross-linking drugs. Phage display was used to isolate peptides that bind the paralog XRCC3. Sequences of binding peptides showed similarity to residues 14 25 of RAD51C protein. Point mutations in this region of RAD51C altered its interaction with both XRCC3 and RAD51B in a two-hybrid system. A synthetic peptide composed of residues 14-25 of RAD51C fused to a membrane transduction sequence [protein transduction domain 4 (PTD4)], inhibited subnuclear assembly of RAD51 recombinase, and sensitized Chinese hamster ovary cells to cisplatin when added to growth medium. These results suggest that residues 14-25 of RAD51C contribute to a "hot spot" used in both XRCC3-RAD51C and RAD51B-RAD51C interactions. Peptide-based inhibition of homologous recombination may prove useful for improving the efficacy of existing cancer therapies. PMID- 15126334 TI - X-linked inhibitor of apoptosis protein (XIAP) is a nonredundant modulator of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis in human cancer cells. AB - Although the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been shown to play an important role in the immunosurveillance of neoplasia, apoptotic factors that modulate the sensitivity of cancer cells to TRAIL are poorly understood. The inhibitor of apoptosis proteins (IAPs) have generated considerable interest as potential targets for cancer therapy, but the lack of a phenotype in X-linked IAP (XIAP) knockout mice has generated speculation that IAP function may be redundant. Using gene targeting technology, we show that disruption of the gene encoding XIAP in human cancer cells did not interfere with basal proliferation, but caused a remarkable sensitivity to TRAIL. These results demonstrate that XIAP is a nonredundant modulator of TRAIL-mediated apoptosis and provide a rationale for XIAP as a therapeutic target. PMID- 15126335 TI - Polymorphisms in DNA double-strand break repair genes and skin cancer risk. AB - UV can cause a wide range of DNA lesions. UVA-induced oxidative DNA damage and blocked DNA replication by UVB-induced photoproducts can lead to double-strand breaks (DSBs). We selected 11 haplotype-tagging single nucleotide polymorphisms in three DSB repair genes XRCC2, XRCC3, and LigaseIV and evaluated their associations with skin cancer risk in a nested case-control study within the Nurses' Health Study [219 melanoma, 286 squamous cell carcinoma (SCC), 300 basal cell carcinoma (BCC), and 873 controls]. We observed that the XRCC3 18085T (241Met) allele and its associated haplotype were significantly inversely associated with the risks of SCC and BCC, whereas the XRCC3 4552C allele along with its associated haplotype and the XRCC2 30833A allele were significantly associated with increased BCC risk. The LigaseIV 4044T and 4062T alleles were associated with decreased BCC risk; two of four haplotypes were significantly associated with altered BCC risk. A trend toward decreased risk of nonmelanoma skin cancer was found in those harboring a greater number of putative low risk alleles (P for trend, 0.05 for SCC, <0.0001 for BCC). The main effects of these genotypes were essentially null for melanoma risk. This study provides evidence to suggest the role of the DSB repair pathway in skin cancer development, especially for BCC. PMID- 15126336 TI - Frequent inactivation of PTEN by promoter hypermethylation in microsatellite instability-high sporadic colorectal cancers. AB - Loss of PTEN tumor suppressor function is observed in tumors of breast, prostate, thyroid, and endometrial origin. Allelic losses in the proximity of the PTEN locus (10q23) also occur in sporadic colorectal cancers (CRCs), but biallelic inactivation of this site has not been frequently demonstrated. We hypothesized that alternative mechanisms of PTEN allelic inactivation, such as promoter hypermethylation, might be operative in CRC and that PTEN inactivation may be related to recognized forms of genomic instability. We characterized a cohort of 273 sporadic CRCs by determining their microsatellite instability (MSI) status. Of these, 146 cancers were examined for PTEN promoter methylation by methylation specific PCR. Mutations at the poly(A)6 repeat sequences in PTEN exons 7 and 8 and deletions at the 10q23 locus were also identified using microsatellite analysis. The presence of PTEN protein was determined by immunostaining, and the results were correlated with the promoter methylation status. We observed that PTEN promoter hypermethylation was a frequent occurrence in MSI-high (MSI-H) tumors (19.1% of MSI-H versus 2.2% of MSI-low/microsatellite stable tumors; P = 0.002). A PTEN mutation or a deletion event was present in 60% of the tumors with promoter region hypermethylation. Hypermethylation of the PTEN promoter correlated significantly with either decreased or complete loss of PTEN protein expression (P = 0.004). This is the first demonstration of PTEN inactivation as a result of promoter hypermethylation in MSI-H sporadic CRCs. These data suggest that this silencing mechanism plays a major role in PTEN inactivation and, in colon cancer, may be more important than either allelic losses or inactivating mutations. The significant correlation of PTEN hypermethylation with MSI-H tumors further suggests that PTEN is an additional important "target" of methylation along with the hMLH1 gene in the evolution of MSI-H CRCs and also confers the "second hit" in the biallelic inactivation mechanism for some proportion of tumors. PMID- 15126337 TI - Apoptotic signaling pathways induced by nitric oxide in human lymphoblastoid cells expressing wild-type or mutant p53. AB - Loss of p53 function by inactivating mutations results in abrogation of NO*induced apoptosis in human lymphoblastoid cells. Here we report characterization of apoptotic signaling pathways activated by NO* in these cells by cDNA microarray expression and immunoblotting. A p53-mediated transcriptional response to NO* was observed in p53-wild-type TK6, but not in closely related p53 mutant WTK1, cells. Several previously characterized p53 target genes were up regulated transcriptionally in TK6 cells, including phosphatase PPM1D (WIP1), oxidoreductase homolog PIG3, death receptor TNFRSF6 (Fas/CD95), and BH3-only proteins BBC3 (PUMA) and PMAIP1 (NOXA). NO* also modulated levels of several gene products in the mitochondria-dependent and death-receptor-mediated apoptotic pathways. Inhibitors of apoptosis proteins X-chromosome-linked inhibitor of apoptosis, cellular inhibitor of apoptosis protein-1, and survivin were significantly down-regulated in TK6 cells, but not in WTK1 cells. Smac release from mitochondria was induced in both cell types, but release of apoptosis inducing factor and endonuclease G was detected only in TK6 cells. Fas/CD95 was increased, and levels of the antiapoptotic proteins Bcl-2 and Bcl-x/L were reduced in TK6 cells. Activation of procaspases 3, 8, 9, and 10, as well as Bid and poly(ADP-ribose) polymerase cleavage, were observed only in TK6 cells. NO* treatment did not alter levels of death receptors 4 and 5, Fas-associated death domain or proapoptotic Bax and Bak proteins in either cell line. Collectively, these data show that NO* exposure activated a complex network of responses leading to p53-dependent apoptosis via both mitochondrial and Fas receptor pathways, which were abrogated in the presence of mutant p53. PMID- 15126338 TI - Clustering of minimal deleted regions reveals distinct genetic pathways of human hepatocellular carcinoma. AB - Systematic scan and statistical analysis of loss of heterozygosity (LOH) has been widely used to define chromosomal aberrations in various cancers for cloning of tumor suppressor genes and for development of prognostic markers. However, the establishment of novel strategies is needed, so that the nonrandom but heterogeneous chromosomal aberration data could provide significant insights into our understanding of molecular pathogenesis of cancers. After comprehensive allelotyping of recurrent allelic losses with 441 highly informative microsatellite markers and overlapping LOH regions on human hepatocellular carcinoma (HCC) chromosomes, 33 minimal deleted regions (MDRs) were revealed. Five and 15 of the 33 MDRs have physical intervals in less than 5 and 10 Mb, respectively, with the smallest MDR9p1 of 2.2 Mb located at 9p21.3-p21.2. Statistical and Kaplan-Meier survival analysis revealed a significant association between the loss of MDR15q1 (15q21.1-q22.2) and the HCC patient survival (adjusted P = 0.033). After cluster analysis of 33 MDRs that represented LOH profiles of each HCC tissue based on clinicopathological features and p53 mutations, two major genetic pathways, low-stage and advanced-stage HCC, were uncovered based on high concordance of MDR clusters. We propose that the definition of genome-wide MDRs on the cancer genome not only narrows down the location of existing tumor suppressor genes to facilitate positional candidate cloning and develop potential prognostic markers after statistical association of MDRs with clinicopathological features but also dissects genetic interactions and pathways of chromosomal aberrations in tumorigenesis. PMID- 15126339 TI - Expression profiling of purified normal human luminal and myoepithelial breast cells: identification of novel prognostic markers for breast cancer. AB - The normal duct-lobular system of the breast is lined by two epithelial cell types, inner luminal secretory cells and outer contractile myoepithelial cells. We have generated comprehensive expression profiles of the two normal cell types, using immunomagnetic cell separation and gene expression microarray analysis. The cell-type specificity was confirmed at the protein level by immunohistochemistry in normal breast tissue. New prognostic markers for survival were identified when the luminal- and myoepithelial-specific molecules were evaluated on breast tumor tissue microarrays. Nuclear expression of luminal epithelial marker galectin 3 correlated with a shorter overall survival in these patients, and the expression of SPARC (osteonectin), a myoepithelial marker, was an independent marker of poor prognosis in breast cancers as a whole. These data provide a framework for the interpretation of breast cancer molecular profiling experiments, the identification of potential new diagnostic markers, and development of novel indicators of prognosis. PMID- 15126340 TI - HOXB13 homeodomain protein suppresses the growth of prostate cancer cells by the negative regulation of T-cell factor 4. AB - In prostate gland, HOXB13 is highly expressed from the embryonic stages to adulthood. However, the function of HOXB13 in normal cell growth and tumorigenesis is not yet known. We investigated the role of HOXB13 and mechanism by which it functions in HOXB13-negative cells. Expression of HOXB13 was forced in HOXB13-negative PC3 prostate cancer cells using a liposome-mediated gene transfer approach. Compared with the control clones, HOXB13-expressing PC3 cells exhibited significant inhibition of in vitro and in vivo cell growth with G1 cell cycle arrest mediated by the suppression of cyclin D1 expression. Because cyclin D1 is mainly regulated by beta-catenin/T-cell factor (TCF), TCF-4 response element was used in a reporter gene transcription assay, demonstrating that HOXB13 significantly inhibits TCF-4-mediated transcriptional activity in both prostate and nonprostate cells. This inhibition occurred in a dose-responsive manner and was specific to TCF-4 response element. Western blot analysis demonstrated that HOXB13 down-regulates the expression of TCF-4 and its responsive genes, c-myc and cyclin D1. HOXB13 also suppressed the activity of natural c-myc promoter. This study suggests that HOXB13, a transcription factor, functions as a cell growth suppressor by negatively regulating the expression of TCF-4, which eventually provides negative signals for cell proliferation. This observation will provide valuable insight into the molecular basis of prostate tumorigenesis. PMID- 15126341 TI - Genome-wide array-based comparative genomic hybridization reveals multiple amplification targets and novel homozygous deletions in pancreatic carcinoma cell lines. AB - Pancreatic carcinomas display highly complex chromosomal abnormalities, including many structural and numerical aberrations. There is ample evidence indicating that some of these abnormalities, such as recurrent amplifications and homozygous deletions, contribute to tumorigenesis by altering expression levels of critical oncogenes and tumor suppressor genes. To increase the understanding of gene copy number changes in pancreatic carcinomas and to identify key amplification/deletion targets, we applied genome-wide array-based comparative genomic hybridization to 31 pancreatic carcinoma cell lines. Two different microarrays were used, one containing 3,565 fluorescence in situ hybridization verified bacterial artificial chromosome clones and one containing 25,468 cDNA clones representing 17,494 UniGene clusters. Overall, the analyses revealed a high genomic complexity, with several copy number changes detected in each case. Specifically, 60 amplicons at 32 different locations were identified, most frequently located within 8q (8 cases), 12p (7 cases), 7q (5 cases), 18q (5 cases), 19q (5 cases), 6p (4 cases), and 8p (4 cases). Amplifications of 8q and 12p were mainly clustered at 8q23-24 and 12p11-12, respectively, whereas amplifications on other chromosome arms were more dispersed. Furthermore, our analyses identified several novel homozygously deleted segments located to 9p24, 9p21, 9q32, 10p12, 10q22, 12q24, and 18q23. The individual complexity and aberration patterns varied substantially among cases, i.e., some cell lines were characterized mainly by high-level amplifications, whereas others showed primarily whole-arm imbalances and homozygous deletions. The described amplification and deletion targets are likely to contain genes important in pancreatic tumorigenesis. PMID- 15126342 TI - An integrated view of copy number and allelic alterations in the cancer genome using single nucleotide polymorphism arrays. AB - Changes in DNA copy number contribute to cancer pathogenesis. We now show that high-density single nucleotide polymorphism (SNP) arrays can detect copy number alterations. By hybridizing genomic representations of breast and lung carcinoma cell line and lung tumor DNA to SNP arrays, and measuring locus-specific hybridization intensity, we detected both known and novel genomic amplifications and homozygous deletions in these cancer samples. Moreover, by combining genotyping with SNP quantitation, we could distinguish loss of heterozygosity events caused by hemizygous deletion from those that occur by copy-neutral events. The simultaneous measurement of DNA copy number changes and loss of heterozygosity events by SNP arrays should strengthen our ability to discover cancer-causing genes and to refine cancer diagnosis. PMID- 15126343 TI - Identification of Sp2 as a transcriptional repressor of carcinoembryonic antigen related cell adhesion molecule 1 in tumorigenesis. AB - Down-regulation of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) tumor suppressor gene expression is common in several malignancies including prostate, colon, and breast cancer. The mechanism that mediates this down-regulation is not known. Here, we report that down-regulation of CEACAM1 expression in prostate cancer cells occurs primarily at the transcriptional level and is mediated by Sp2, a member of the Sp family of transcription factors. Sp2 binds to the CEACAM1 promoter in vitro and in vivo, and transient overexpression of Sp2 down-regulates endogenous CEACAM1 expression in normal prostate epithelial cells. Sp2 appears to repress CEACAM1 gene expression by recruiting histone deacetylase activity to the CEACAM1 promoter. In human prostate cancer specimens, Sp2 expression is high in prostate cancer cells but low in normal prostate epithelial cells and is inversely correlated with CEACAM1 expression. Our studies show that transcriptional repression by Sp2 represents one mechanism by which CEACAM1 tumor suppressor gene is down-regulated in prostate cancer. PMID- 15126344 TI - Role of the retinoblastoma pathway in senescence triggered by repression of the human papillomavirus E7 protein in cervical carcinoma cells. AB - Repression of the endogenous human papillomavirus (HPV) type 18 E7 gene in HeLa cervical carcinoma cells by the bovine papillomavirus E2 transcription factor activates the retinoblastoma (Rb) pathway and induces cells to undergo senescence. To determine whether activation of the Rb pathway is responsible for senescence in response to HPV18 E7 repression, we tested the ability of wild-type and mutant E7 proteins to affect the activity of the Rb pathway and to modulate senescence in these cells. Enforced expression of the wild-type HPV16 E7 protein prevented Rb activation in response to E2 expression and impaired senescence. Importantly, there was an absolute correlation between the ability of mutant E7 proteins to inactivate the Rb pathway and to inhibit senescence in HeLa cells. Similar results were obtained in HT-3 cervical carcinoma cells. These results provide strong genetic evidence that activation of the Rb pathway is required for senescence in response to E7 repression. Hence, continuous neutralization of the Rb pathway by the E7 protein is required to maintain the proliferation of cervical carcinoma cells. Similarly, our results indicate that activation of the Rb pathway can prevent apoptosis induced by repression of the HPV18 E6 gene in HeLa cells. PMID- 15126345 TI - Identification and characterization of a novel gene, C13orf25, as a target for 13q31-q32 amplification in malignant lymphoma. AB - The amplification at 13q31-q32 has been reported in not only hematopoietic malignancies but also in other solid tumors. We identified previously frequent amplification of chromosomal band 13q31-q32 in 70 cases of diffuse large B-cell lymphoma patients by conventional comparative genomic hybridization analysis. In an attempt to identify a candidate gene within this region, we used array comparative genomic hybridization and fluorescent in situ hybridization to map the 13q31-q32 amplicon. We then screened the 65 expressed sequence tags and Glypican 5 (GPC5) by reverse transcription-PCR and Northern blotting. As a result, we identified a novel gene, designated Chromosome 13 open reading frame 25 (C13orf25), which was overexpressed in B-cell lymphoma cell lines and diffuse large B-cell lymphoma patients with 13q31-q32 amplifications. However, GPC5, which has been reported to be a target gene for 13q31-q32 amplification, was truncated in one cell line, Rec1, possessing the amplification, and its expression in various cell lines with amplification at 13q31-q32 was not significantly different from that in other cell lines without amplification, suggesting that GPC5 is not likely to be the candidate gene. Additional analysis identified two major transcripts in the C13orf25 gene. The two transcripts A and B predicted open reading frames of 32 and 70-amino acid polypeptides, respectively. The former has been reported as bA121J7.2, which is conserved among species. Transcript-B also contained seven mature microRNAs in its untranslated region. These results suggest that the C13orf25 gene is the most likely candidate gene for the 13q31-q32 amplicon found in hematopoietic malignancies. PMID- 15126346 TI - Deficiencies in mouse Myh and Ogg1 result in tumor predisposition and G to T mutations in codon 12 of the K-ras oncogene in lung tumors. AB - Oxidative DNA damage is unavoidably and continuously generated by oxidant byproducts of normal cellular metabolism. The DNA damage repair genes, mutY and mutM, prevent G to T mutations caused by reactive oxygen species in Escherichia coli, but it has remained debatable whether deficiencies in their mammalian counterparts, Myh and Ogg1, are directly involved in tumorigenesis. Here, we demonstrate that deficiencies in Myh and Ogg1 predispose 65.7% of mice to tumors, predominantly lung and ovarian tumors, and lymphomas. Remarkably, subsequent analyses identified G to T mutations in 75% of the lung tumors at an activating hot spot, codon 12, of the K-ras oncogene, but none in their adjacent normal tissues. Moreover, malignant lung tumors were increased with combined heterozygosity of Msh2, a mismatch repair gene involved in oxidative DNA damage repair as well. Thus, oxidative DNA damage appears to play a causal role in tumorigenesis, and codon 12 of K-ras is likely to be an important downstream target in lung tumorigenesis. The multiple oxidative repair genes are required to prevent mutagenesis and tumor formation. The mice described here provide a valuable model for studying the mechanisms of oxidative DNA damage in tumorigenesis and investigating preventive or therapeutic approaches. PMID- 15126347 TI - Microarray-based screening for molecular markers in medulloblastoma revealed STK15 as independent predictor for survival. AB - Medulloblastoma, a primitive neuroectodermal tumor of the cerebellum, is one of the most common central nervous system malignancies of childhood. Despite aggressive multimodal therapy, including surgery, irradiation, and chemotherapy, 5-year survival rates have only approached 50-60%. To identify potential candidate genes that predict for overall survival (OS), we performed a gene expression profiling analysis in 35 newly diagnosed medulloblastoma neoplasms. Subsequently, the nine most promising candidate genes were analyzed by immunohistochemistry and fluorescence in situ hybridization on tumor tissue microarrays representing a series of 180 tumors. We found 54 genes in which expression levels predicted for unfavorable survival in medulloblastoma. In line with the gene expression profiling analysis, a positive staining for STK15 (P = 0.0006), stathmin 1 (P = 0.001), and cyclin D1 (P = 0.03) was associated with an unfavorable OS, whereas cyclin B1, DAXX, Ki-67, MYC, NRAS, and p53 showed no statistical significant effect. In comparison to clinically defined parameters such as gender, age, metastatic stage, extent of tumor resection, application of chemotherapy, and tumor grade, positive staining for STK15 was identified as an independent prognostic factor for OS (P = 0.026). Moreover, additional gene copy numbers of MYC (P = 0.003) and STK15 (P = 0.05) predicted for poor survival. The combination of gene expression profiling with tissue microarray experiments allowed the identification of a series of candidate genes that predicts for survival in medulloblastoma. Of the results highlighted by the various data analysis procedures, genes associated with cell proliferation (cyclin D1), transcription (MYC), and especially mitosis (stathmin 1, STK15) appear particularly intriguing with respect to medulloblastoma pathomechanism. PMID- 15126348 TI - Galectin-1(L11A) predicted from a computed galectin-1 farnesyl-binding pocket selectively inhibits Ras-GTP. AB - Ras biological activity necessitates membrane anchorage that depends on the Ras farnesyl moiety and is strengthened by Ras/galectin-1 interactions. We identified a hydrophobic pocket in galectin-1, analogous to the Cdc42 geranylgeranyl-binding cavity in RhoGDI, possessing homologous isoprenoid-binding residues, including the critical L11, whose RhoGDI L77 homologue changes dramatically on Cdc42 binding. By substituting L11A, we obtained a dominant interfering galectin-1 that possessed normal carbohydrate-binding capacity but inhibited H-Ras GTP-loading and extracellular signal-regulated kinase activation, dislodged H-Ras(G12V) from the cell membrane, and attenuated H-Ras(G12V) fibroblast transformation and PC12 cell neurite outgrowth. Thus, independently of carbohydrate binding, galectin-1 cooperates with Ras, whereas galectin-1(L11A) inhibits it. PMID- 15126349 TI - Human CYP1B1 is regulated by estradiol via estrogen receptor. AB - Human cytochrome P450 (CYP) 1B1 is a key enzyme in the metabolism of 17beta estradiol (E2). CYP1B1 is mainly expressed in endocrine-regulated tissues, such as mammary, uterus, and ovary. Because many CYP enzymes are likely to be induced by the substrates themselves, we examined whether the human CYP1B1 expression is regulated by E2 in the present study. Real-time reverse transcription-PCR analysis revealed that treatment with 10 nM E2 for 12 h induced CYP1B1 mRNA expression in estrogen receptor (ER)-positive MCF-7 cells. Luciferase reporter assays using MCF-7 cells showed a significant transactivation up to 7-fold by E2 with a reporter plasmid containing a region from -152 to +25 of the human CYP1B1 gene. A computer-assisted homology search indicated a putative estrogen response element (ERE) between -63 and -49 in the CYP1B1 promoter region. Specific binding of ERalpha to the putative ERE was demonstrated by chromatin immunoprecipitation assays and gel shift analyses. With reporter plasmids containing the wild or mutated putative ERE on the CYP1B1 gene and the wild or mutated ERalpha expression vectors, luciferase assays using Ishikawa cells demonstrated that the putative ERE and ERalpha are essential for the transactivation by E2. Because endometrial tissue is highly regulated by estrogens, the expression pattern of CYP1B1 protein in human endometrial specimens was examined by immunohistochemistry. The staining of CYP1B1 was stronger in glandular epithelial cells during a proliferative phase than those during a secretory phase, consistent with the pattern of estrogen secretion. These findings clearly indicated that the human CYP1B1 is regulated by estrogen via ERalpha. Because 4 hydroxylation of estrogen by CYP1B1 leads to decrease of the estrogenic activity but the produced metabolite is toxicologically active, our findings suggest a clinical significance in the estrogen-regulated CYP1B1 expression for the homeostasis of estrogens as well as estrogen-dependent carcinogenesis. PMID- 15126350 TI - Myc-transformed epithelial cells down-regulate clusterin, which inhibits their growth in vitro and carcinogenesis in vivo. AB - Effective treatment of malignant carcinomas requires identification of proteins regulating epithelial cell proliferation. To this end, we compared gene expression profiles in murine colonocytes and their c-Myc-transformed counterparts, which possess enhanced proliferative potential. A surprisingly short list of deregulated genes included the cDNA for clusterin, an extracellular glycoprotein without a firmly established function. We had previously demonstrated that in organs such as skin, clusterin expression is restricted to differentiating but not proliferating cell layers, suggesting a possible negative role in cell division. Indeed, its transient overexpression in Myc-transduced colonocytes decreased cell accumulation. Furthermore, clusterin was down regulated in rapidly dividing human keratinocytes infected with a Myc-encoding adenovirus. Its knockdown via antisense RNA in neoplastic epidermoid cells enhanced proliferation. Finally, recombinant human clusterin suppressed, in a dose-dependent manner, DNA replication in keratinocytes and other cells of epithelial origin. Thus, clusterin appears to be an inhibitor of epithelial cell proliferation in vitro. To determine whether it also affects neoplastic growth in vivo, we compared wild-type and clusterin-null mice with respect to their sensitivity to 7, 12-dimethylbenz(a)anthracene /12-Otetradecanoylphorbol-13 acetate (DMBA/TPA)-induced skin carcinogenesis. We observed that the mean number of papillomas/mouse was higher in clusterin-null animals. Moreover, these papillomas did not regress as readily as in wild-type mice and persisted beyond week 35. The rate of progression toward squamous cell carcinoma was not altered, although those developing in clusterin-null mice were on average better differentiated. These data suggest that clusterin not only suppresses epithelial cell proliferation in vitro but also interferes with the promotion stage of skin carcinogenesis. PMID- 15126351 TI - RNA interference-mediated knockdown of DNA methyltransferase 1 leads to promoter demethylation and gene re-expression in human lung and breast cancer cells. AB - DNA methyltransferase 1 (DNMT1) is required to maintain DNA methylation patterns in mammalian cells, and is thought to be the predominant maintenance methyltransferase gene. Recent studies indicate that inhibiting DNMT1 protein expression may be a useful approach for understanding the role of DNA methylation in tumorigenesis. To this end, we used RNA interference to specifically down regulate DNMT1 protein expression in NCI-H1299 lung cancer and HCC1954 breast cancer cells. RNA interference-mediated knockdown of DNMT1 protein expression resulted in >80% reduction of promoter methylation in RASSF1A, p16(ink4A), and CDH1 in NCI-H1299; and RASSF1A, p16(ink4A), and HPP1 in HCC1954; and re expression of p16(ink4A), CDH1, RASSF1A, and SEMA3B in NCI-H1299; and p16(ink4A), RASSF1A, and HPP1 in HCC1954. By contrast, promoter methylation and lack of gene expression was maintained when these cell lines were treated with control small interfering RNAs. The small interfering RNA treatment was stopped and 17 days later, all of the sequences showed promoter methylation and gene expression was again dramatically down-regulated, indicating the tumor cells still were programmed for these epigenetic changes. We saw no effects on soft agar colony formation of H1299 cells 14 days after DNMT1 knockdown indicating that either these genes are not functioning as tumor suppressors under these conditions, or that more prolonged knockdown or other factors are also required to inhibit the malignant phenotype. These results provide direct evidence that loss of DNMT1 expression abrogates tumor-associated promoter methylation and the resultant silencing of multiple genes implicated in the pathogenesis of human lung and breast cancer. PMID- 15126352 TI - Extensive chromosomal breaks are induced by tamoxifen and estrogen in DNA repair deficient cells. AB - Tamoxifen (TAM) possesses antiestrogen activity and is widely used for the treatment or prevention of breast cancer. However, it is also carcinogenic in human uterus and rat liver, highlighting the profound complexity of its actions. To explore the molecular mechanisms of TAM-induced mutagenesis, we analyzed the effects of this drug on gene-disrupted chicken B lymphocyte (DT40) clones deficient in various DNA repair pathways. Rad18, Rev3, and Polkappa are involved in translesion DNA synthesis (TLS), which facilitates recovery from replication blocks on damaged template strands. DT40 cells deficient in TLS were found to be hypersensitive to TAM, exhibiting an increase in chromosomal breaks. Furthermore, these mutants were also hypersensitive to 4-hydroxyestradiol, a physiological metabolite of estrogen. These data suggest a contribution of TLS to the prevention of chromosomal breaks by TAM and estrogen, and they therefore indicate that such error-prone DNA synthesis underlies mutagenesis induced by these agents. PMID- 15126353 TI - Identification of heme oxygenase-1 as a novel BCR/ABL-dependent survival factor in chronic myeloid leukemia. AB - Chronic myeloid leukemia (CML) is a stem cell disease in which BCR/ABL promotes the survival of leukemic cells. Heme oxygenase-1 (HO-1) is an inducible stress protein that catalyzes the degradation of heme and has recently been implicated in the regulation of growth and survival of various neoplastic cells. In the present study, we analyzed the expression and role of HO-1 in CML cells. As assessed by Northern and Western blot analysis as well as immunostaining, primary CML cells were found to express HO-1 mRNA and the HO-1 protein in a constitutive manner. Exposure of these cells to the BCR/ABL tyrosine kinase inhibitor STI571 resulted in decreased expression of HO-1 mRNA and protein. In addition, BCR/ABL was found to up-regulate HO-1 promoter activity, mRNA levels, and protein levels in Ba/F3 cells. To investigate the role of HO-1 for survival of primary CML cells, the HO-1 inducer hemin was used. Hemin-induced expression of HO-1 was found to protect CML cells from STI571-induced cell death. In addition, inhibition of HO-1 by zinc-(II)-deuteroporphyrin-IX-2,4-bisethyleneglycol resulted in a substantial decrease of cell viability. Furthermore, overexpression of HO-1 in the CML-derived cell line K562 was found to counteract STI571-induced apoptosis. Together, our data identify HO-1 as a novel BCR/ABL-driven survival molecule and potential target in leukemic cells in patients with CML. The pathogenetic and clinical implications of this observation remain to be elucidated. PMID- 15126354 TI - High-resolution magnetic resonance imaging of disparities in the transcapillary transfer rates in orthotopically inoculated invasive breast tumors. AB - In vivo mapping of the transcapillary fluxes in tumors can help predict the efficacy of delivery of blood-borne anticancer drugs. These fluxes are primarily affected by the vascular permeability and the pressure gradients across the blood vessels' walls. We describe herein high-resolution dynamic contrast-enhanced magnetic resonance imaging of the influx and outflux transcapillary transfer rates in vivo in invasive MDA-MB-231 tumors orthotopically inoculated in severe combined immunodeficient mice. The tumors were noted for rapid growth, impaired drainage of fluid, and subsequent formation of cysts. Consequently, the time evolution of the contrast enhancement, induced by i.v. injection of Gadolinium diethylene-triamine-penta-acetate, exhibited two distinct patterns: transcapillary transfer in the cellular regions and simple diffusion in the cyst fluid. Both processes were analyzed at pixel resolution applying to each a physiological model and a corresponding algorithm. In the cellular region, the influx and outflux transcapillary transfer rates decreased during tumor growth; however, an increased disparity between the transfer constants was observed, with the outflux rate exceeding the influx rate. This quantitative spatial and temporal mapping of this disparity can provide a means to assess the physiological barriers to tracer delivery. It is hypothesized that both the increased disparity in transcapillary transfer rates and impaired fluid drainage in these tumors could arise from the development of interstitial hypertension. PMID- 15126356 TI - Activation of Akt-1 (PKB-alpha) can accelerate ErbB-2-mediated mammary tumorigenesis but suppresses tumor invasion. AB - Elevated expression of Akt-1 (PKBalpha) has been noted in a significant percentage of primary human breast cancers. Another frequent event in the genesis of human breast cancers is amplification and overexpression of the ErbB-2 receptor tyrosine kinase, an event which is associated with activation of Akt-1. To directly assess the importance of Akt-1 activation in ErbB-2 mammary tumor progression, we interbred separate strains of transgenic mice carrying mouse mammary tumor virus/activated Akt-1 and mouse mammary tumor virus/activated ErbB 2 to derive progeny that coexpress the transgenes in the mammary epithelium. Female transgenic mice coexpressing activated Akt-1 and ErbB-2 develop multifocal mammary tumors with a significantly shorter latency period than mice expressing activated ErbB-2 alone. This dramatic acceleration of mammary tumor progression correlates with enhanced cellular proliferation, elevated Cyclin D1 protein levels, and phosphorylation of retinoblastoma protein. These bitransgenic mammary tumors also exhibit lower levels of invasion into the surrounding tissue and more differentiated phenotypes. Consistent with these observations, female mice coexpressing activated Akt-1 and ErbB-2 developed significantly fewer metastatic lesions than the activated ErbB-2 strain alone. Taken together, these observations suggest that activation of Akt-1 during ErbB-2-induced mammary tumorigenesis may have opposing effects on tumor growth and metastatic progression. PMID- 15126355 TI - Activation of peroxisome proliferator-activated receptor delta stimulates the proliferation of human breast and prostate cancer cell lines. AB - The nuclear receptor peroxisome proliferator-activated receptor delta [PPARdelta/beta (NR1C2)] has been implicated in colorectal carcinogenesis by various molecular genetic observations. These observations have recently been supported by studies of activation of PPARdelta by pharmacological agents. Here we present the first report of the stimulation of breast and prostate cancer cell growth using PPARdelta selective agonists. Activation of PPARdelta with compound F stimulated proliferation in breast (T47D, MCF7) and prostate (LNCaP, PNT1A) cell lines, which are responsive to sex hormones. Conversely, we have found that several steroid-independent cell lines, including colon lines, were unresponsive to compound F. These findings were confirmed with an additional high-affinity PPARdelta agonist, GW501516. Conditional expression of PPARdelta in MCF7 Tet-On cells resulted in a doxycycline-enhanced response to GW501516, thus providing direct genetic evidence for the role of PPARdelta in the proliferative response to this drug. Activation of PPARdelta in T47D cells resulted in increased expression of the proliferation marker Cdk2 and also vascular endothelial growth factor alpha (VEGFalpha) and its receptor, FLT-1, thus, suggesting that PPARdelta may initiate an autocrine loop for cellular proliferation and possibly angiogenesis. Consistent with this hypothesis, we demonstrated a pro proliferative effect of GW501516 on human umbilical vein endothelial cell cultures and found that GW501516 also regulated the expression of VEGFalpha and FLT-1 in these cells. Our observations provide the first evidence that activation of PPARdelta can result in increased growth in breast and prostate cancer cell lines and primary endothelial cells and supports the possibility that PPARdelta antagonists may be of therapeutic value in the treatment of breast and prostate cancer. PMID- 15126357 TI - The phosphorylation of EphB2 receptor regulates migration and invasion of human glioma cells. AB - Eph receptor tyrosine kinases and their ligands, ephrins, mediate neurodevelopmental processes such as boundary formation, axon guidance, vasculogenesis, and cell migration. We determined the expression profiles of the Eph family members in five glioma cell lines under migrating and nonmigrating conditions. EphB2 mRNA was overexpressed in all five during migration (1.2-2.8 fold). We found abundant EphB2 protein as well as strong phosphorylation of EphB2 in migrating U87 cells. Confocal imaging showed EphB2 localized in lamellipodia of motile U87 cells. Treatment with ephrin-B1/Fc chimera stimulated migration and invasion of U87, whereas treatment with a blocking EphB2 antibody significantly inhibited migration and invasion. Forced expression of EphB2 in U251 cells stimulated cell migration and invasion and diminished adhesion concomitant with the tyrosine phosphorylation of EphB2. U251 stably transfected with EphB2 showed more scattered and more pronounced invasive growth in an ex vivo rat brain slice. In human brain tumor specimens, EphB2 expression was higher in glioblastomas than in low-grade astrocytomas or normal brain; patterns of phosphorylated EphB2 matched the expression levels. Laser capture microdissection of invading glioblastoma cells revealed elevated EphB2 mRNA (1.5-3.5-fold) in 7 of 7 biopsy specimens. Immunohistochemistry demonstrated EphB2 localization primarily in glioblastoma cells (56 of 62 cases) and not in normal brain. This is the first demonstration that migrating glioblastoma cells overexpress EphB2 in vitro and in vivo; glioma migration and invasion are promoted by activation of EphB2 or inhibited by blocking EphB2. Dysregulation of EphB2 expression or function may underlie glioma invasion. PMID- 15126358 TI - Interleukin-1beta regulates angiopoietin-1 expression in human endothelial cells. AB - Angiopoietin (Ang)-1 is an important regulator of endothelial cell (EC) survival and stabilization. Ang-1 exerts its biological effects by binding to the EC specific tyrosine kinase receptor Tie-2, and initiates intracellular signaling in ECs. However, regulatory mechanisms for endothelial Ang-1 expression have not been completely elucidated. In this study, we investigated the effects of angiogenic cytokines and growth factors on Ang-1 expression in human umbilical vein ECs (HUVECs). Northern blot analysis was performed after HUVECs were exposed to interleukin-1beta (IL-1beta), tumor necrosis factor-alpha, platelet-derived growth factor-BB, insulin-like growth factor-1, or vascular endothelial growth factor (VEGF). Both IL-1beta and tumor necrosis factor-alpha caused marked down regulation of Ang-1 mRNA levels at 4 h with a further decrease observed at 24 h. Using signaling inhibitors, we identified the P38 pathway as the pathway that mediates IL-1beta down-regulation of Ang-1. Furthermore, treatment of cells with IL-1beta indirectly (via down-regulation of Ang-1) led to a decrease in Tie-2 autophosphorylation levels in HUVECs. We previously demonstrated that IL-1beta regulates VEGF expression in tumor cells. This observation was confirmed in ECs in the present study. Because pericytes play a role in regulating EC function, we also determined whether IL-1beta would also down-regulate Ang-1 in human vascular smooth muscle cells. Similar to our findings in HUVECs, we found that IL-1beta decreased Ang-1 expression in human vascular smooth muscle cells. Direct effects of IL-1beta on angiogenesis were investigated by use of an in vivo Gelfoam angiogenesis assay in which IL-1beta produced a significant increase in vessel counts (P = 0.0189). These results suggest that IL-1beta indirectly regulates angiogenesis by modulating the expression of Ang-1. IL-1beta may trigger a proangiogenic response by decreasing Ang-1 levels in ECs and pericytes and up regulating VEGF in ECs and tumor cells. PMID- 15126359 TI - Expression of focal adhesion kinase in acute myeloid leukemia is associated with enhanced blast migration, increased cellularity, and poor prognosis. AB - Focal adhesion kinase (FAK) is a nonreceptor tyrosine kinase playing an important role in cell motility and survival. However, very little is known about FAK in normal and leukemic myeloid cells. In this study, FAK protein expression and mRNA were detected in 25 of 60 cases (42%) of acute myeloid leukemia (AML). Whereas FAK was expressed in 46% of CD34+ AML cells, it was not detected in normal purified CD34+ cells. Conversely, the FAK homologue proline-rich tyrosine kinase 2 (PYK2) was found to be expressed both in normal and leukemic myeloid cells. When expressed, FAK displayed phosphorylation on Tyr-397, an important step for its activation. Moreover, FAK expression was correlated with the phosphorylation of PYK2 on Tyr-881, a critical site for the PYK2 function in cell migration. FAK+ AML cells displayed significantly higher migration capacities and resistance to daunorubicin, compared with FAK- cells. The implication of FAK in both cell motility and drug resistance was demonstrated by small interfering RNA experiments with the FAK-positive KG1 cell line. However, adhesion on fibronectin efficiently protected FAK- AML cells from daunorubicin-mediated killing, suggesting that cellular adhesion mediated-drug resistance is not mediated by FAK. Finally, in a retrospective cohort of 60 AML patients, FAK expression was significantly correlated with high blast cell count, early death, and shorter survival rate. Altogether, this study shows that FAK is aberrantly expressed and activated in about half of the cases of AML and suggests that FAK may contribute to the regulation of AML cell transit from the marrow to blood compartment and that it may influence clinical outcome. PMID- 15126360 TI - Tumor-specific low molecular weight forms of cyclin E induce genomic instability and resistance to p21, p27, and antiestrogens in breast cancer. AB - The deregulated expression of cyclin E as measured by the overexpression of its low molecular weight (LMW) isoforms is a powerful predictor of poor outcome in patients with breast cancer. The mechanism by which these LMW forms give tumor cells a growth advantage is not known and is the subject of this article. In this article, we provide the pathobiological mechanisms of how these LMW forms are involved in disease progression. Specifically, we show that overexpression of the LMW forms of cyclin E but not the full-length form in MCF-7 results in (a) their hyperactivity because of increased affinity for cdk2 and resistance to inhibition by the cyclin-dependent kinase inhibitors p21 and p27, (b) resistance to the growth inhibiting effects of antiestrogens, and (c) chromosomal instability. Lastly, tumors from breast cancer patients overexpressing the LMW forms of cyclin E are polyploid in nature and are resistant to endocrine therapy. Collectively, the biochemical and functional differences between the full-length and the LMW isoforms of cyclin E provide a molecular mechanism for the poor clinical outcome observed in breast cancer patients harboring tumors expressing high levels of the LMW forms of cyclin E. These properties of the LMW forms cyclin E suggest that they are not just surrogate markers of poor outcome but bona fide mediators of aggressive disease and potential therapeutic targets for patients whose tumors overexpress these forms. PMID- 15126361 TI - Endothelin-1 is a critical mediator of myogenic tone in tumor arterioles: implications for cancer treatment. AB - Although derived from the host tissue, the tumor vasculature is under the influence of the tumor microenvironment and needs to adapt to the resistance to blood flow inherent to the dynamics of tumor growth. Such vascular remodeling can offer selective targets to pharmacologically modulate tumor perfusion and thereby improve the efficacy of conventional anticancer treatments. Radiotherapy and chemotherapy can, indeed, take advantage of a better tumor oxygenation and drug delivery, respectively, both partly dependent on the tumor blood supply. Here, we showed that isolated tumor arterioles mounted in a pressure myograph have the ability, contrary to size-matched healthy arterioles, to contract in response to a transluminal pressure increase. This myogenic tone was exquisitely dependent on the endothelin-1 pathway because it was completely abolished by the selective endothelin receptor A (ETA) antagonist BQ123. This selectivity was additionally supported by the large increase in endothelin-1 abundance in tumors and the higher density of the ETA receptors in tumor vessels. We also documented by using laser Doppler microprobes and imaging that administration of the ETA antagonist led to a significant increase in tumor blood flow, whereas the perfusion in control healthy tissue was not altered. Finally, we provided evidence that acute administration of the ETA antagonist could significantly stimulate tumor oxygenation, as determined by electron paramagnetic resonance oximetry, and increase the efficacy of low-dose, clinically relevant fractionated radiotherapy. Thus, blocking the tumor-selective increase in the vascular endothelin-1/ETA pathway led us to unravel an important reserve of vasorelaxation that can be exploited to selectively increase tumor response to radiotherapy. PMID- 15126362 TI - Radiation to stromal fibroblasts increases invasiveness of pancreatic cancer cells through tumor-stromal interactions. AB - Radiotherapy represents a major treatment option for patients with pancreatic cancer, but recent evidence suggests that radiation can promote invasion and metastasis of cancer cells. Interactions between cancer cells and surrounding stromal cells may play an important role in aggressive tumor progression. In the present study, we investigated the invasive phenotype of pancreatic cancer cells in response to coculture with irradiated fibroblasts. Using in vitro invasion assay, we demonstrated that coculture with nonirradiated fibroblasts significantly increased the invasive ability of pancreatic cancer cells and, surprisingly, the increased invasiveness was further accelerated when they were cocultured with irradiated fibroblasts. The hepatocyte growth factor (HGF) secretion from fibroblasts remained unchanged after irradiation, whereas exposure of pancreatic cancer cells to supernatant from irradiated fibroblasts resulted in increased phosphorylation of c-Met (HGF receptor) and mitogen-activated protein kinase activity, possibly or partially via increased expression of c-Met. We also demonstrated that scattering of pancreatic cancer cells was accelerated by the supernatant from irradiated fibroblasts. The enhanced invasiveness of pancreatic cancer cells induced by coculture with irradiated fibroblasts was completely blocked by NK4, a specific antagonist of HGF. These data suggest that invasive potential of certain pancreatic cancer cells is enhanced by soluble mediator(s) released from irradiated fibroblasts possibly through up-regulation of c-Met expression/phosphorylation and mitogen-activated protein kinase activity in pancreatic cancer cells. Our present findings further support the potential use of NK4 during radiotherapy for patients with pancreatic cancer. PMID- 15126363 TI - In vivo treatment with CPT-11 leads to differentiation of neuroblastoma xenografts and topoisomerase I alterations. AB - Topoisomerase I inhibitors, such as CPT-11, are potent anticancer drugs against neuroblastoma (NB). Differentiating agents, such as retinoids, improve the survival of children with metastatic NB. To characterize the biological effects associated with exposure to CPT-11 in vivo, athymic mice bearing a human NB xenograft, named IGR-NB8 and characterized as an immature NB with poor prognostic markers, were treated with CPT-11. Prolonged stable disease was observed, resulting in an overall tumor growth delay of 115 days. During treatment, tumors differentiated into ganglioneuroblastomas (GGNB), which reverted into an immature phenotype when treatment was discontinued. In contrast, 13-cis retinoic acid failed to induce differentiation of IGR-NB8 in vivo. Tumor differentiation was associated with decreased N-myc expression, induction of p73 expression in the perinuclear area and cytoplasm, and a dramatic 35-fold decrease in topoisomerase I (topo I) catalytic activity. The full-length Mr 100,000 topo I protein was present in both pre and post-treatment immature NB xenografts. In contrast, differentiated GGNBs did not contain the Mr 100,000 protein but an intense Mr 48,000 topo I fragment. Furthermore, redistribution of the Mr 48,000 and 68,000 forms to the cytoplasm was observed in differentiated tumors. The same pattern of topo I expression and catalytic activity was observed in NBs and GGNBs obtained from pediatric patients. Our data suggest that prolonged in vivo exposure to CPT 11 induces differentiation of NB xenografts, which is associated with truncation of the topo I enzyme, relocation of the degraded forms to the cytoplasm, and decreased catalytic activity. PMID- 15126364 TI - Celecoxib can prevent tumor growth and distant metastasis in postoperative setting. AB - Much evidence suggests that an inflammatory condition provides a microenvironment favorable for tumor growth. One of the main components in the healing wound is the induction of cyclooxygenase-2 (COX-2) and prostaglandins, and many solid tumors have been known to overexpress COX-2. The present study investigated the relationship between surgical wounds and tumor growth and the roles of COX-2 and inflammatory reaction in this microenvironment. We created surgical wounds in syngeneic mice for the implantation of SCC VII murine cancer cell line. Accelerated tumor growth and increased angiogenesis by surgical wounds were clearly observed in C3H/HeJ mice with SCC VII tumor. The COX-2 expression of peritumoral tissues and leukocyte infiltration partly explained the accelerated tumor growth, especially in the early phase after surgical wounding. Celecoxib had a significantly suppressive effect on tumor growth, angiogenesis, and metastasis in tumor-implanted mice with surgical wounds. This tumor-suppressive action of celecoxib did not show any noticeable side effects on the late wound healing and on the gastrointestinal tracts. Prophylactic use of the drug can be advocated in many clinical situations, such as residual tumors or contamination of surgical fields by tumor cells. PMID- 15126366 TI - Characterization of the interaction of ingenol 3-angelate with protein kinase C. AB - Ingenol 3-angelate (I3A) is one of the active ingredients in Euphorbia peplus, which has been used in traditional medicine. Here, we report the initial characterization of I3A as a protein kinase C (PKC) ligand. I3A bound to PKC alpha in the presence of phosphatidylserine with high affinity; however, under these assay conditions, little PKC isoform selectivity was observed. PKC isoforms did show different sensitivity and selectivity for down-regulation by I3A and phorbol 12-myristate 13-acetate (PMA) in WEHI-231, HOP-92, and Colo-205 cells. In all of the three cell types, I3A inhibited cell proliferation with somewhat lower potency than did PMA. In intact CHO-K1 cells, I3A was able to translocate different green fluorescent protein-tagged PKC isoforms, visualized by confocal microscopy, with equal or higher potency than PMA. PKC-delta in particular showed a different pattern of translocation in response to I3A and PMA. I3A induced a higher level of secretion of the inflammatory cytokine interleukin 6 compared with PMA in the WEHI-231 cells and displayed a marked biphasic dose-response curve for the induction. I3A was unable to cause the same extent of association of the C1b domain of PKC-delta with lipids, compared with PMA or the physiological regulator diacylglycerol, and was able to partially block the association induced by these agents, measured by surface plasmon resonance. The in vitro kinase activity of PKC-alpha induced by I3A was lower than that induced by PMA. The novel pattern of behavior of I3A makes it of great interest for further evaluation. PMID- 15126365 TI - Inducible release of TRAIL fusion proteins from a proapoptotic form for tumor therapy. AB - Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) can selectively kill neoplastic cells and control of its activity could enhance tumor therapy. We have developed means to control the secretion of a novel recombinant (r) TRAIL fusion protein using a viral protease. This system uses the endoplasmic reticulum (ER) as a storage depot for rTRAIL, because TRAIL acts by binding to its cognate receptors on the cell surface. We have engineered two TRAIL variants: (a) a secretable form that enhances apoptosis via a bystander effect; and (b) an ER targeted TRAIL that is retained in the ER until selectively released by the viral protease. Gene delivery can be monitored in vivo by systemic administration of a near infrared fluorescent (NIRF) probe activated by the protease. This study serves as a template for design of recombinant proteins to enhance and control apoptosis of tumor cells via specific viral proteases and for use of viral proteases as in vivo reporters for cancer therapy. PMID- 15126368 TI - Syncytia induction enhances the oncolytic potential of vesicular stomatitis virus in virotherapy for cancer. AB - Vesicular stomatitis virus (VSV) selectively replicates in tumor but not in normal cells and is being developed as an oncolytic agent for cancer therapy. Here we report the construction of a recombinant VSV capable of inducing syncytia formation between tumor cells through membrane fusion at neutral pH, which led to enhanced oncolytic properties against multifocal hepatocellular carcinoma (HCC) in the livers of immunocompetent rats. Recombinant VSV vectors were constructed by insertion into their genome a transcription unit expressing a control or fusion protein derived from Newcastle disease virus. In vitro characterization of the recombinant fusogenic VSV vector on human and rat HCC cells showed extensive syncytia formation and significantly enhanced cytotoxic effects. In vivo, administration of fusogenic VSV into the hepatic artery of Buffalo rats bearing syngeneic multifocal HCC lesions in their livers resulted in syncytia formation exclusively within the tumors, and there was no collateral damage to the neighboring hepatic parenchyma. The fusogenic VSV also conferred a significant survival advantage over a nonfusogenic control virus in the treated animals (P = 0.0078, log-rank test). The results suggest that fusogenic VSV can be developed into an effective and safe therapeutic agent for cancer treatment in patients, including those with multifocal HCC in the liver. PMID- 15126367 TI - Ubiquitination and proteasomal degradation of nucleophosmin-anaplastic lymphoma kinase induced by 17-allylamino-demethoxygeldanamycin: role of the co-chaperone carboxyl heat shock protein 70-interacting protein. AB - Nucleophosmin-anaplastic lymphoma kinase (NPM-ALK) is a constitutively active fusion tyrosine kinase involved in lymphomagenesis of human anaplastic large cell lymphomas (ALCL), the maturation and activity of which depend on the association with the heat shock protein (hsp) 90 protein chaperone. Targeting hsp90 by the ansamycins geldanamycin and 17-allyl-amino-demethoxygeldanamycin (17-AAG) promotes degradation of several proteins through the ubiquitin-proteasome pathway, including oncogenic Raf, v-Src, erbB2, and BCR-ABL. We have previously shown that 17-AAG prevents hsp90/NPM-ALK complex formation and fosters NPM-ALK turnover, perhaps through its association with the hsp70 chaperone. Here, we show that inhibition of the proteasome activity by the potent and specific compound pyrazylcarbonyl-Phe-Leu-boronate (PS-341) blocks 17-AAG-induced down-regulation of NPM-ALK, which becomes detergent-insoluble and relocates into ubiquitin-rich perinuclear vesicles that represent aggregated polyubiquitinated forms of the protein. Kinase activity was not mandatory for proteasomal degradation of NPM ALK, because kinase-defective NPM-ALK was even more rapidly degraded upon 17-AAG treatment. Prolonged exposure to the proteasome inhibitor was shown to trigger caspase-3-mediated apoptosis in proliferating ALCL cells at nanomolar concentrations. However, we verified that the accumulation of detergent-insoluble NPM-ALK in ALCL cells was not a spurious consequence of PS341-committed apoptosis, because caspase inhibitors prevented poly(ADP-ribose) polymerase cleavage whereas they did not affect partitioning of aggregated NPM-ALK. In line with these observations, the carboxyl hsp70-interacting ubiquitin ligase (CHIP), was shown to increase basal ubiquitination and turnover of NPM-ALK kinase, supporting a mechanism whereby NPM-ALK proceeds rapidly toward hsp70-assisted ubiquitin-dependent proteasomal degradation, when chaperoning activity of hsp90 is prohibited by 17-AAG. PMID- 15126369 TI - Soluble interleukin-13Ralpha2 decoy receptor inhibits Hodgkin's lymphoma growth in vitro and in vivo. AB - Recent studies have demonstrated that the malignant Reed-Sternberg cells of Hodgkin's lymphoma (HL) secrete and are responsive to interleukin (IL)-13. We hypothesized that overexpression of a soluble IL-13 decoy receptor (sIL 13Ralpha2) via adenoviral-mediated gene transfer would inhibit IL-13-induced Reed Sternberg cell proliferation. Western blot and ELISA analysis verified expression of sIL-13Ralpha2 in cell lysates and supernatants of AdsIL-13Ralpha2-transduced COS-7 cells. Treatment of two IL-13-responsive HL-derived cell lines, HDLM-2 and L-1236, with AdsIL-13Ralpha2-conditioned medium, resulted in the inhibition of cell proliferation, and down-regulated the phosphorylation of signal transducer and activator of transcription 6 (STAT6), an important mediator of IL-13 signaling. i.v. delivery of AdsIL-13Ralpha2 in NOD/SCID mice with s.c. implanted HDLM-2 cells delayed tumor onset and growth while enhancing survival compared with control mice. Intratumoral administration of AdsIL-13Ralpha2 led to the regression or stabilization of established tumors and was associated with diminished STAT6 phosphorylation. Our data demonstrate that AdsIL-13Ralpha2 can suppress HL growth in vitro and in vivo. PMID- 15126370 TI - Antitumor activity of a kinesin inhibitor. AB - Several members of the kinesin family of microtubule motor proteins play essential roles in mitotic spindle function and are potential targets for the discovery of novel antimitotic cancer therapies. KSP, also known as HsEg5, is a kinesin that plays an essential role in formation of a bipolar mitotic spindle and is required for cell cycle progression through mitosis. We identified a potent inhibitor of KSP, CK0106023, which causes mitotic arrest and growth inhibition in several human tumor cell lines. Here we show that CK0106023 is an allosteric inhibitor of KSP motor domain ATPase with a Ki of 12 nM. Among five kinesins tested, CK0106023 was specific for KSP. In tumor-bearing mice, CK0106023 exhibited antitumor activity comparable to or exceeding that of paclitaxel and caused the formation of monopolar mitotic figures identical to those produced in cultured cells. KSP was most abundant in proliferating human tissues and was absent from cultured postmitotic neurons. These findings are the first to demonstrate the feasibility of targeting mitotic kinesins for the treatment of cancer. PMID- 15126372 TI - Disruption of cancer cell replication by alternating electric fields. AB - Low-intensity, intermediate-frequency (100-300 kHz), alternating electric fields, delivered by means of insulated electrodes, were found to have a profound inhibitory effect on the growth rate of a variety of human and rodent tumor cell lines (Patricia C, U-118, U-87, H-1299, MDA231, PC3, B16F1, F-98, C-6, RG2, and CT-26) and malignant tumors in animals. This effect, shown to be nonthermal, selectively affects dividing cells while quiescent cells are left intact. These fields act in two modes: arrest of cell proliferation and destruction of cells while undergoing division. Both effects are demonstrated when such fields are applied for 24 h to cells undergoing mitosis that is oriented roughly along the field direction. The first mode of action is manifested by interference with the proper formation of the mitotic spindle, whereas the second results in rapid disintegration of the dividing cells. Both effects, which are frequency dependent, are consistent with the computed directional forces exerted by these specific fields on charges and dipoles within the dividing cells. In vivo treatment of tumors in C57BL/6 and BALB/c mice (B16F1 and CT-26 syngeneic tumor models, respectively), resulted in significant slowing of tumor growth and extensive destruction of tumor cells within 3-6 days. These findings demonstrate the potential applicability of the described electric fields as a novel therapeutic modality for malignant tumors. PMID- 15126371 TI - Adenovirus vector-mediated in vivo gene transfer of OX40 ligand to tumor cells enhances antitumor immunity of tumor-bearing hosts. AB - OX40 ligand (OX40L), the ligand for OX40 on activated CD4+ T cells, has adjuvant properties for establishing effective T-cell immunity, a potent effector arm of the immune system against cancer. The hypothesis of this study is that in vivo genetic engineering of tumor cells to express OX40L will stimulate tumor-specific T cells by the OX40L-OX40 engagement, leading to an induction of systemic antitumor immunity. To investigate this hypothesis, s.c. established tumors of three different mouse cancer cells (B16 melanoma, H-2b; Lewis lung carcinoma, H 2b; and Colon-26 colon adenocarcinoma, H-2d) were treated with intratumoral injection of a recombinant adenovirus vector expressing mouse OX40L (AdOX40L). In all tumor models tested, treatment of tumor-bearing mice with AdOX40L induced a significant suppression of tumor growth along with survival advantages in the treated mice. The in vivo AdOX40L modification of tumors evoked tumor-specific cytotoxic T lymphocytes in the treated host correlated with in vivo priming of T helper 1 immune responses in a tumor-specific manner. Consistent with the finding, the antitumor effect provided by intratumoral injection of AdOX40L was completely abrogated in a CD4+ T cell-deficient or CD8+ T cell-deficient condition. In addition, ex vivo AdOX40L-transduced B16 cells also elicited B16 specific cytotoxic T lymphocyte responses, and significantly suppressed the B16 tumor growth in the immunization-challenge experiment. All of these results support the concept that genetic modification of tumor cells with a recombinant OX40L adenovirus vector may be of benefit in cancer immunotherapy protocols. PMID- 15126373 TI - Expression, up-regulation, and transport activity of the multidrug-resistance protein Abcg2 at the mouse blood-brain barrier. AB - The breast cancer resistance protein (BCRP/ABCG2) is, like P-glycoprotein (P-gp), a member of the ABC family of drug transporters. These proteins actively transport various anticancer drugs from cells, causing multidrug resistance. The physiological expression of P-gp/ABCB1 at the blood-brain barrier (BBB) effectively restricts the brain uptake of many antitumor drugs by mediating their active efflux from the brain to the blood vessel lumen. However, little is known about the function of Abcg2 at the BBB in vivo. We used in situ brain perfusion to measure the uptake of two known Abcg2 substrates, prazosin and mitoxantrone, and the nonsubstrate vinblastine by the brains of wild-type and P-gp-deficient mutant mdr1a(-/-) mice with or without the P-gp/Abcg2 inhibitor GF120918 or the P gp inhibitor PSC833. P-gp had no effect on the brain transport of prazosin and mitoxantrone at the mouse BBB, but wild-type and P-gp-deficient mouse brains perfused with GF120918 or a high concentration of prazosin showed carrier mediated effluxes of prazosin and mitoxantrone from the brain that did not involve P-gp. In contrast, the brain uptake of vinblastine was restricted only by P-gp and not by Abcg2 at the BBB. The amounts of abcg2 mRNA in cortex homogenates and capillary-enriched fractions of wild-type and mdr1a(-/-) mouse brains were measured by real-time quantitative reverse transcription-PCR. There was approximately 700-times more abcg2 mRNA in brain microvessels than in the cortex of the wild-type mice, confirming that Abcg2 plays an important role at the BBB. There was also approximately 3 times more abcg2 mRNA in the microvessels from P gp-deficient mutant mouse brains than in the microvessels of wild-type mouse brains. These findings confirm that Abcg2 is a physiological transporter at the BBB that restricts the permeability of the brain to its substrates in vivo. Lastly, the defective P-gp in the mutant mdr1a(-/-) mice was associated with increased abcg2 mRNA at the BBB and a greater export of prazosin and mitoxantrone from the brain, as measured in the P-gp-deficient mice versus the wild-type mice. PMID- 15126374 TI - A retinoid-related molecule that does not bind to classical retinoid receptors potently induces apoptosis in human prostate cancer cells through rapid caspase activation. AB - Synthetic retinoid-related molecules, such as N-(4-hydroxyphenyl)retinamide (fenretinide) and 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalene carboxylic acid (CD437) induce apoptosis in a variety of malignant cells. The mechanism(s) of action of these compounds does not appear to involve retinoic acid receptors (RARs) and retinoid X receptors (RXRs), although some investigators disagree with this view. To clarify whether some retinoid-related molecules can induce apoptosis without involving RARs and/or RXRs, we used 4-[3-(1-heptyl-4,4-dimethyl 2-oxo-1,2,3,4-tetrahydroquinolin-6-yl)-3-oxo-E-propenyl] benzoic acid (AGN193198) that neither binds effectively to RARs and RXRs nor transactivates in RAR- and RXR-mediated reporter assays. AGN193198 potently induced apoptosis in prostate, breast, and gastrointestinal carcinoma cells and in leukemia cells. AGN193198 also abolished growth (by 50% at 130-332 nM) and induced apoptosis in primary cultures established from prostatic carcinoma (13 patients) and gastrointestinal carcinoma (1 patient). Apoptosis was induced rapidly, as indicated by mitochondrial depolarization and DNA fragmentation. Molecular events provoked by AGN193198 included activation of caspase-3, -8, -9, and -10 (by 4-6 h) and the production of BID/p15 (by 6 h). These findings show that caspase-mediated induction of apoptosis by AGN193198 is RAR/RXR-independent and suggest that this compound may be useful in the treatment of prostate cancer. PMID- 15126375 TI - Selective preservation of pemetrexed pharmacological activity in HeLa cells lacking the reduced folate carrier: association with the presence of a secondary transport pathway. AB - A methotrexate (MTX)-resistant HeLa subline (R5), developed in this laboratory, with impaired transport due to a genomic deletion of the reduced folate carrier (RFC) was only 2-fold resistant to pemetrexed (PMX), but 200- and 400-fold resistant to raltitrexed (ZD1694) and N(alpha)-(-4-amino-4-deoxypteroyl)-N(delta) hemiphthaloyl-1-ornithine (PT523), respectively, compared with parental HeLa cells when grown with 2 microM folic acid. When folic acid was replaced with the more physiological 25 nM 5-formyltetrahydrofolate, R5 cells were 2-fold collaterally sensitive to PMX but still 40- and 200-fold resistant to ZD1694 and PT523, respectively. Sensitivity to PT523 and PMX could be completely restored, and sensitivity to ZD1694 nearly restored, by transfection of RFC cDNA into R5 cells, indicating that the defect in drug transport was the only, or major, factor in resistance. The preserved PMX activity in R5 cells could not be related to the very low expression of folate receptors. Rather, retained PMX activity in R5 cells was associated with residual transport by another process that exhibits good affinity for PMX (Kt = 12 microM) with much lower affinities for ZD1694, MTX, and PT523 (Kis of approximately 90, 100, and 250 microM, respectively). PMX transported by this route was rapidly converted to higher polyglutamates and, when grown with 25 nM 5-formyl-tetrahydrofolate, the rate of formation of these derivatives and their net accumulation in R5 cells was comparable to that of wild type cells. These data suggest that selective preservation of PMX pharmacological activity in RFC-null R5 cells is due, in part, to partial preservation of transport by secondary process with a higher affinity for PMX than the other antifolates evaluated. PMID- 15126376 TI - A novel synthetic inhibitor of CDC25 phosphatases: BN82002. AB - CDC25 dual-specificity phosphatases are essential regulators that dephosphorylate and activate cyclin-dependent kinase/cyclin complexes at key transitions of the cell cycle. CDC25 activity is currently considered to be an interesting target for the development of new antiproliferative agents. Here we report the identification of a new CDC25 inhibitor and the characterization of its effects at the molecular and cellular levels, and in animal models. BN82002 inhibits the phosphatase activity of recombinant human CDC25A, B, and C in vitro. It impairs the proliferation of tumoral cell lines and increases cyclin-dependent kinase 1 inhibitory tyrosine phosphorylation. In synchronized HeLa cells, BN82002 delays cell cycle progression at G1-S, in S phase and at the G2-M transition. In contrast, BN82002 arrests U2OS cell cycle mostly in the G1 phase. Selectivity of this inhibitor is demonstrated: (a) by the reversion of the mitotic-inducing effect observed in HeLa cells upon CDC25B overexpression; and (b) by the partial reversion of cell cycle arrest in U2OS expressing CDC25. We also show that BN82002 reduces growth rate of human tumor xenografts in athymic nude mice. BN82002 is a original CDC25 inhibitor that is active both in cell and animal models. This greatly reinforces the interest in CDC25 as an anticancer target. PMID- 15126377 TI - Different procarcinogenic potentials of lymphocyte subsets in a transgenic mouse model of chronic hepatitis B. AB - The immune response to hepatitis viruses is believed to be involved in the development of chronic hepatitis; however, its pathogenetic potential has not been clearly defined. The current study, using a transgenic mouse model of chronic hepatitis B, was designed to determine the relative contributions of the immune cell subsets to the progression of liver disease that induces hepatocellular carcinogenesis. Hepatitis B virus transgenic mice were adoptively transferred with CD4+ and CD8+ T cell-enriched or -depleted and B cell-depleted splenocytes obtained from hepatitis B surface antigen-primed, syngeneic nontransgenic donors. The resultant liver disease, hepatocyte apoptosis, regeneration, and tumor development were assessed and compared with the manifestations in mice that had received unfractionated spleen cells. Transfer of CD8(+)-enriched splenocytes caused prolonged disease kinetics, and a marked increase in the extent of hepatocyte apoptosis and regeneration. In 12 of 14 mice the transfer resulted in multiple hepatocellular carcinomas (HCCs) comparable with the manifestations seen in the mice transferred with total splenocytes. In contrast, mice that had received CD4(+)-enriched cells demonstrated lower levels of liver disease and developed fewer incidences of HCC (4 of 17). The experiment also revealed that all of the groups of mice complicated with HCC developed comparable mean numbers and sizes of tumors. B-cell depletion had no effect on disease kinetics in this model. Taken together, these results demonstrate that the pathogenetic events induced by CD8+ T-cell subset are primarily responsible for the induction of chronic liver disease that increases tumor incidence, suggesting their potential in triggering the process of hepatocarcinogenesis. PMID- 15126378 TI - Suppression of prostate carcinogenesis by dietary supplementation of celecoxib in transgenic adenocarcinoma of the mouse prostate model. AB - Epidemiological studies and clinical observations suggest that nonsteroidal anti inflammatory drugs and certain selective cyclooxygenase (COX)-2 inhibitors may reduce the relative risk of clinically evident prostate cancer. This prompted us to investigate the chemopreventive potential of celecoxib, a selective COX-2 inhibitor, against prostate carcinogenesis in a transgenic adenocarcinoma of the mouse prostate (TRAMP) model. Similar to prostate cancer in humans, prostate malignancies in TRAMP mice progress from precursor intraepithelial lesions, to invasive carcinoma that metastasizes to lymph nodes, liver, lungs, and occasionally to bone. The basal enzyme activity and protein expression of COX-2 is significantly higher (>4-fold) in the dorsolateral prostate of TRAMP mice up to 24 weeks of age compared with their nontransgenic littermates. Eight-week-old TRAMP mice were randomly divided and fed either control diet (AIN 76A) or a custom prepared AIN 76A diet containing 1500-ppm celecoxib ad libitum for 24 weeks, a dosage that would compare with the normal recommended dose for the treatment of human disease. Studies from two independent experiments, each consisting of 10 mice on test, showed that the cumulative incidence of prostate cancer development at 32 weeks of age in animals fed with AIN 76A diet was 100% (20 of 20) as observed by tumor palpation, whereas 65% (13 of 20), 35% (7 of 20), and 20% (4 of 20) of the animals exhibited distant site metastases to lymph nodes, lungs, and liver. Celecoxib supplementation to TRAMP mice from 8-32 weeks of age exhibited significant reduction in tumor development (5 of 20) with no signs of metastasis. Celecoxib feeding resulted in a significant decrease in prostate (56%; P < 0.0003) and genitourinary weight (48%; P < 0.008). Sequential magnetic resonance imaging analysis of celecoxib-fed mice documented lower prostate volume compared with the AIN 76A-fed group. Histopathological examination of celecoxib-fed animals showed reduced proliferation, and down modulation of COX-2 and prostaglandin E2 levels in the dorsolateral prostate and plasma, respectively. These results correlated with retention of antimetastasis markers, viz E-cadherin, and alpha- and beta-catenin, along with a significant decrease in vascular endothelial growth factor protein expression. Celecoxib supplementation also resulted in enhanced in vivo apoptosis in the prostate as monitored by several techniques including a recently perfected technique of 99mTc labeled annexin V in live animals followed by phosphor imaging. One striking observation in an additional study was that celecoxib feeding to mice with established tumors (16 weeks of age) significantly improved their overall survival (P = 0.014), compared with AIN 76A-fed group. Our findings suggest that celecoxib may be useful in chemoprevention of prostate cancer. PMID- 15126379 TI - Caffeine inhibits cell proliferation by G0/G1 phase arrest in JB6 cells. AB - Caffeine is a major biologically active constituent in coffee and tea. Because caffeine has been reported to inhibit carcinogenesis in UVB-exposed mice, the cancer-preventing effect of caffeine has attracted considerable attention. In the present study, the effect of caffeine in quiescent (G0 phase) cells was investigated. Pretreatment with caffeine suppressed cell proliferation in a dose dependent manner 36 h after addition of fetal bovine serum as a cell growth stimulator. Analysis by flow cytometry showed that caffeine suppressed cell cycle progression at the G0/G1 phase, i.e., 18 h after addition of fetal bovine serum, the percentages of cells in G0/G1 phase in 1 mM caffeine-treated cells and in caffeine-untreated cells were 61.7 and 29.0, respectively. The percentage of cells in G0/G1 phase at 0 h was 75.5. Caffeine inhibited phosphorylation of retinoblastoma protein at Ser780 and Ser807/Ser811, the sites where retinoblastoma protein has been reported to be phosphorylated by cyclin-dependent kinase 4 (cdk4). Furthermore, caffeine inhibited the activation of the cyclin D1 cdk4 complex in a dose-dependent manner. However this compound did not directly inhibit the activity of this complex. In addition, caffeine did not affect p16INK4 or p27Kip1 protein levels, but inhibited the phosphorylation of protein kinase B (Akt) and glycogen synthase kinase 3beta. Our results showed that caffeine suppressed the progression of quiescent cells into the cell cycle. The inhibitory mechanism may be due to the inhibition of cell growth signal-induced activation of cdk4, which may be involved in the inhibition of carcinogenesis in vivo. PMID- 15126380 TI - Proceedings of the Oxygen Homeostasis/Hypoxia Meeting. PMID- 15126381 TI - Frontiers of mutagenesis and DNA repair: a workshop. PMID- 15126382 TI - Two lessons from the interface of genetics and medicine. PMID- 15126383 TI - Strategies for avoiding reinventing the precollege education and outreach wheel. AB - The National Science Foundation's recent mandate that all Principal Investigators address the broader impacts of their research has prompted an unprecedented number of scientists to seek opportunities to participate in precollege education and outreach. To help interested geneticists avoid duplicating efforts and make use of existing resources, we examined several precollege genetics, genomics, and biotechnology education efforts and noted the elements that contributed to their success, indicated by program expansion, participant satisfaction, or participant learning. Identifying a specific audience and their needs and resources, involving K-12 teachers in program development, and evaluating program efforts are integral to program success. We highlighted a few innovative programs to illustrate these findings. Challenges that may compromise further development and dissemination of these programs include absence of reward systems for participation in outreach as well as lack of training for scientists doing outreach. Several programs and institutions are tackling these issues in ways that will help sustain outreach efforts while allowing them to be modified to meet the changing needs of their participants, including scientists, teachers, and students. Most importantly, resources and personnel are available to facilitate greater and deeper involvement of scientists in precollege and public education. PMID- 15126384 TI - Recombination and migration of Cryphonectria hypovirus 1 as inferred from gene genealogies and the coalescent. AB - Genealogy-based methods were used to estimate migration of the fungal virus Cryphonectria hypovirus 1 between vegetative compatibility types of the host fungus, Cryphonectria parasitica, as a means of estimating horizontal transmission within two host populations. Vegetative incompatibility is a self/non-self recognition system that inhibits virus transmission under laboratory conditions but its effect on transmission in nature has not been clearly demonstrated. Recombination within and among different loci in the virus genome restricted the genealogical analyses to haplotypes with common mutation and recombinational histories. The existence of recombination necessitated that we also use genealogical approaches that can take advantage of both the mutation and recombinational histories of the sample. Virus migration between populations was significantly restricted. In contrast, estimates of migration between vegetative compatibility types were relatively high within populations despite previous evidence that transmission in the laboratory was restricted. The discordance between laboratory estimates and migration estimates from natural populations highlights the challenges in estimating pathogen transmission rates. Genealogical analyses inferred migration patterns throughout the entire coalescent history of one viral region in natural populations and not just recent patterns of migration or laboratory transmission. This application of genealogical analyses provides markedly stronger inferences on overall transmission rates than laboratory estimates do. PMID- 15126385 TI - RuvAB and RecG are not essential for the recovery of DNA synthesis following UV induced DNA damage in Escherichia coli. AB - Ultraviolet light induces DNA lesions that block the progression of the replication machinery. Several models speculate that the resumption of replication following disruption by UV-induced DNA damage requires regression of the nascent DNA or migration of the replication machinery away from the blocking lesion to allow repair or bypass of the lesion to occur. Both RuvAB and RecG catalyze branch migration of three- and four-stranded DNA junctions in vitro and are proposed to catalyze fork regression in vivo. To examine this possibility, we characterized the recovery of DNA synthesis in ruvAB and recG mutants. We found that in the absence of either RecG or RuvAB, arrested replication forks are maintained and DNA synthesis is resumed with kinetics that are similar to those in wild-type cells. The data presented here indicate that RecG- or RuvAB catalyzed fork regression is not essential for DNA synthesis to resume following arrest by UV-induced DNA damage in vivo. PMID- 15126386 TI - EXO1 plays a role in generating type I and type II survivors in budding yeast. AB - Telomerase-defective budding yeast cells escape senescence by using homologous recombination to amplify telomeric or subtelomeric structures. Similarly, human cells that enter senescence can use homologous recombination for telomere maintenance, when telomerase cannot be activated. Although recombination proteins required to generate telomerase-independent survivors have been intensively studied, little is known about the nucleases that generate the substrates for recombination. Here we demonstrate that the Exo1 exonuclease is an initiator of the recombination process that allows cells to escape senescence and become immortal in the absence of telomerase. We show that EXO1 is important for generating type I survivors in yku70delta mre11delta cells and type II survivors in tlc1delta cells. Moreover, in tlc1delta cells, EXO1 seems to contribute to the senescence process itself. PMID- 15126387 TI - EXO1 contributes to telomere maintenance in both telomerase-proficient and telomerase-deficient Saccharomyces cerevisiae. AB - Previous work in budding yeast has indicated that telomeres are protected, at least in part, from the action of Exo1, which degrades the C-rich strand of partially uncapped telomeres. To explore this further, we examined the consequences of Exo1-mediated activity in strains that lacked Ku, telomerase, or both. Loss of Exo1 partially rescued the telomere length defect in a yku80delta strain, demonstrating that exonuclease action can directly contribute to telomere shortening. The rapid loss of inviability displayed by a yku80delta est2delta strain was also partially alleviated by an exo1delta mutation, further supporting the proposal that Exo1 is one target of the activities that normally protect wild type telomeres. Conversely, however, Exo1 activity was also capable of enhancing telomere function and consequently cell proliferation, by contributing to a telomerase-independent pathway for telomere maintenance. The recovery of recombination-dependent survivors that arose in a yku80delta est2delta strain was partially dependent on Exo1 activity. Furthermore, the types of recombination events that facilitate telomerase-independent survival were influenced by Exo1 activity, in both est2delta and yku80delta est2delta strains. These data demonstrate that Exo1 can make either positive or negative contributions to telomere function and cell viability, depending on whether telomerase or recombination is utilized to maintain telomere function. PMID- 15126388 TI - Saccharomyces cerevisiae SSD1-V confers longevity by a Sir2p-independent mechanism. AB - The SSD1 gene of Saccharomyces cerevisiae is a polymorphic locus that affects diverse cellular processes including cell integrity, cell cycle progression, and growth at high temperature. We show here that the SSD1-V allele is necessary for cells to achieve extremely long life span. Furthermore, addition of SSD1-V to cells can increase longevity independently of SIR2, although SIR2 is necessary for SSD1-V cells to attain maximal life span. Past studies of yeast aging have been performed in short-lived ssd1-d strain backgrounds. We propose that SSD1-V defines a previously undescribed pathway affecting cellular longevity and suggest that future studies on longevity-promoting genes should be carried out in long lived SSD1-V strains. PMID- 15126389 TI - Yeast MPH1 gene functions in an error-free DNA damage bypass pathway that requires genes from Homologous recombination, but not from postreplicative repair. AB - The MPH1 gene from Saccharomyces cerevisiae, encoding a member of the DEAH family of proteins, had been identified by virtue of the spontaneous mutator phenotype of respective deletion mutants. Genetic analysis suggested that MPH1 functions in a previously uncharacterized DNA repair pathway that protects the cells from damage-induced mutations. We have now analyzed genetic interactions of mph1 with a variety of mutants from different repair systems with respect to spontaneous mutation rates and sensitivities to different DNA-damaging agents. The dependence of the mph1 mutator phenotype on REV3 and REV1 and the synergy with mutations in base and nucleotide excision repair suggest an involvement of MPH1 in error-free bypass of lesions. However, although we observed an unexpected partial suppression of the mph1 mutator phenotype by rad5, genetic interactions with other mutations in postreplicative repair imply that MPH1 does not belong to this pathway. Instead, mutations from the homologous recombination pathway were found to be epistatic to mph1 with respect to both spontaneous mutation rates and damage sensitivities. Determination of spontaneous mitotic recombination rates demonstrated that mph1 mutants are not deficient in homologous recombination. On the contrary, in an sgs1 background we found a pronounced hyperrecombination phenotype. Thus, we propose that MPH1 is involved in a branch of homologous recombination that is specifically dedicated to error-free bypass. PMID- 15126390 TI - The putative RNA helicase Dbp6p functionally interacts with Rpl3p, Nop8p and the novel trans-acting Factor Rsa3p during biogenesis of 60S ribosomal subunits in Saccharomyces cerevisiae. AB - Ribosome biogenesis requires at least 18 putative ATP-dependent RNA helicases in Saccharomyces cerevisiae. To explore the functional environment of one of these putative RNA helicases, Dbp6p, we have performed a synthetic lethal screen with dbp6 alleles. We have previously characterized the nonessential Rsa1p, whose null allele is synthetically lethal with dbp6 alleles. Here, we report on the characterization of the four remaining synthetic lethal mutants, which reveals that Dbp6p also functionally interacts with Rpl3p, Nop8p, and the so-far uncharacterized Rsa3p (ribosome assembly 3). The nonessential Rsa3p is a predominantly nucleolar protein required for optimal biogenesis of 60S ribosomal subunits. Both Dbp6p and Rsa3p are associated with complexes that most likely correspond to early pre-60S ribosomal particles. Moreover, Rsa3p is co immunoprecipitated with protA-tagged Dbp6p under low salt conditions. In addition, we have established a synthetic interaction network among factors involved in different aspects of 60S-ribosomal-subunit biogenesis. This extensive genetic analysis reveals that the rsa3 null mutant displays some specificity by being synthetically lethal with dbp6 alleles and by showing some synthetic enhancement with the nop8-101 and the rsa1 null allele. PMID- 15126391 TI - Role of the nuclease activity of Saccharomyces cerevisiae Mre11 in repair of DNA double-strand breaks in mitotic cells. AB - The Rad50:Mre11:Xrs2 (RMX) complex functions in repair of DNA double-strand breaks (DSBs) by recombination and nonhomologous end-joining (NHEJ) and is also required for telomere stability. The Mre11 subunit exhibits nuclease activities in vitro, but the role of these activities in repair in mitotic cells has not been established. In this study we have performed a comparative study of three mutants (mre11-D16A, -D56N, and -H125N) previously shown to have reduced nuclease activities in vitro. In ends-in and ends-out chromosome recombination assays using defined plasmid and oligonucleotide DNA substrates, mre11-D16A cells were as deficient as mre11 null strains, but defects were small in mre11-D56N and H125N mutants. mre11-D16A cells, but not the other mutants, also displayed strong sensitivity to ionizing radiation, with residual resistance largely dependent on the presence of the partially redundant nuclease Exo1. mre11-D16A mutants were also most sensitive to the S-phase-dependent clastogens hydroxyurea and methyl methanesulfonate but, as previously observed for D56N and H125N mutants, were not defective in NHEJ. Importantly, the affinity of purified Mre11-D16A protein for Rad50 and Xrs2 was indistinguishable from wild type and the mutant protein formed complexes with equivalent stoichiometry. Although the role of the nuclease activity has been questioned in previous studies, the comparative data presented here suggest that the nuclease function of Mre11 is required for RMX-mediated recombinational repair and telomere stabilization in mitotic cells. PMID- 15126392 TI - The selective values of alleles in a molecular network model are context dependent. AB - Classical quantitative genetics has applied linear modeling to the problem of mapping genotypic to phenotypic variation. Much of this theory was developed prior to the availability of molecular biology. The current understanding of the mechanisms of gene expression indicates the importance of nonlinear effects resulting from gene interactions. We provide a bridge between genetics and gene network theories by relating key concepts from quantitative genetics to the parameters, variables, and performance functions of genetic networks. We illustrate this methodology by simulating the genetic switch controlling galactose metabolism in yeast and its response to selection for a population of individuals. Results indicate that genes have heterogeneous contributions to phenotypes and that additive and nonadditive effects are context dependent. Early cycles of selection suggest strong additive effects attributed to some genes. Later cycles suggest the presence of strong context-dependent nonadditive effects that are conditional on the outcomes of earlier selection cycles. A single favorable allele cannot be consistently identified for most loci. These results highlight the complications that can arise with the presence of nonlinear effects associated with genes acting in networks when selection is conducted on a population of individuals segregating for the genes contributing to the network. PMID- 15126393 TI - The external amino acid signaling pathway promotes activation of Stp1 and Uga35/Dal81 transcription factors for induction of the AGP1 gene in Saccharomyces cerevisiae. AB - Yeast cells respond to the presence of amino acids in their environment by inducing transcription of several amino acid permease genes including AGP1, BAP2, and BAP3. The signaling pathway responsible for this induction involves Ssy1, a permease-like sensor of external amino acids, and culminates with proteolytic cleavage and translocation to the nucleus of the zinc-finger proteins Stp1 and Stp2, the lack of which abolishes induction of BAP2 and BAP3. Here we show that Stp1-but not Stp2-plays an important role in AGP1 induction, although significant induction of AGP1 by amino acids persists in stp1 and stp1 stp2 mutants. This residual induction depends on the Uga35/Dal81 transcription factor, indicating that the external amino acid signaling pathway activates not only Stp1 and Stp2, but also another Uga35/Dal81-dependent transcriptional circuit. Analysis of the AGP1 gene's upstream region revealed that Stp1 and Uga35/Dal81 act synergistically through a 21-bp cis-acting sequence similar to the UAS(AA) element previously found in the BAP2 and BAP3 upstream regions. Although cells growing under poor nitrogen-supply conditions display much higher induction of AGP1 expression than cells growing under good nitrogen-supply conditions, the UAS(AA) itself is totally insensitive to nitrogen availability. Nitrogen-source control of AGP1 induction is mediated by the GATA factor Gln3, likely acting through adjacent 5'-GATA-3' sequences, to amplify the positive effect of UAS(AA). Our data indicate that Stp1 may act in combination with distinct sets of transcription factors, according to the gene context, to promote induction of transcription in response to external amino acids. The data also suggest that Uga35/Dal81 is yet another transcription factor under the control of the external amino acid sensing pathway. Finally, the data show that the TOR pathway mediating global nitrogen control of transcription does not interfere with the external amino acid signaling pathway. PMID- 15126394 TI - The fluffy gene of Neurospora crassa is necessary and sufficient to induce conidiophore development. AB - The fl (fluffy) gene of Neurospora crassa encodes a binuclear zinc cluster protein that regulates the production of asexual spores called macroconidia. Two other genes, acon-2 and acon-3, play major roles in controlling development. fl is induced specifically in differentiating tissue during conidiation and acon-2 plays a role in this induction. We examined the function of fl by manipulating its level of expression in wild-type and developmental mutant strains. Increasing expression of fl from a heterologous promoter in a wild-type genetic background is sufficient to induce conidiophore development. Elevated expression of fl leads to induction of development of the acon-2 mutant in nitrogen-starved cultures, but does not bypass the conidiation defect of the acon-3 mutant. These findings indicate that fl acts downstream of acon-2 and upstream of acon-3 in regulating gene expression during development. The eas, con-6, and con-10 genes are induced at different times during development. Morphological changes induced by artificially elevated fl expression in the absence of environmental cues were correlated with increased expression of eas, but not con-6 or con-10. Thus, although inappropriate expression of fl in vegetative hyphae is sufficient to induce conidial morphogenesis, complete reconstitution of development leading to the formation of mature conidia may require environmental signals to regulate fl activity and/or appropriate induction of fl expression in the developing conidiophore. PMID- 15126395 TI - Mating within the meiotic tetrad and the maintenance of genomic heterozygosity. AB - Mating among the products of a single meiosis (automixis or meiotic parthenogenesis) is found in diverse groups of plant, animal, and fungal taxa. Restoration of the diploid stage is often strictly controlled and brings together products separated at the first meiotic division. Despite apparent similarities to diploid selfing, the theoretical prediction is that heterozygosity should be maintained on all chromosomes when it is linked to the centromeres and thus also segregates at the first meiotic division. Using the fungus Microbotryum, we directly test this prediction by linear tetrad analysis. The patterns of meiotic segregation for chromosome size variation (electrophoretic karyotypes) and PCR products (AFLP procedures) were determined for Microbotryum lineages native to North America and Europe. Our data reveal a surprisingly dynamic genome that is rich in heterozygosity and where size-dimorphic autosomes are common. The genetic variation agrees with the prediction of centromere-linked heterozygosity. This was observed to the greatest extent in the lineage of Microbotryum native to North America where there was consistent first-division segregation and independent assortment of multiple linkage groups. The data also show properties that distinguish the fungal sex chromosomes from the autosomes in both lineages of Microbotryum. We describe a scenario where the mating system of automixis with first-division restitution is the result of feedback mechanisms to control exposure of genetic load. PMID- 15126396 TI - The evolution of the Drosophila sex-determination pathway. AB - The molecular complexity of the Drosophila somatic sex-determination pathway poses formidable intellectual challenges for attempts to explain its evolutionary origins. Here we present a reconstruction of how this regulatory cascade might have evolved in a step-by-step fashion. We illustrate how mutations in genes, which were already part of the pathway or were recruited as new regulators of the pathway, were favored by sexual selection acting on the discriminatory sex determining signal. This allows us to explain the major features of the pathway, including multiple promoter sites, alternative splicing patterns, autoregulation, and stop codons. Our hypothesis is built on the available data from Drosophila and other insect species, and we point out where it is amenable to further experimental and comparative tests. PMID- 15126397 TI - Construction of transgenic Drosophila by using the site-specific integrase from phage phiC31. AB - The phiC31 integrase functions efficiently in vitro and in Escherichia coli, yeast, and mammalian cells, mediating unidirectional site-specific recombination between its attB and attP recognition sites. Here we show that this site-specific integration system also functions efficiently in Drosophila melanogaster in cultured cells and in embryos. Intramolecular recombination in S2 cells on transfected plasmid DNA carrying the attB and attP recognition sites occurred at a frequency of 47%. In addition, several endogenous pseudo attP sites were identified in the fly genome that were recognized by the integrase and used as substrates for integration in S2 cells. Two lines of Drosophila were created by integrating an attP site into the genome with a P element. phiC31 integrase injected into embryos as mRNA functioned to promote integration of an attB containing plasmid into the attP site, resulting in up to 55% of fertile adults producing transgenic offspring. A total of 100% of these progeny carried a precise integration event at the genomic attP site. These experiments demonstrate the potential for precise genetic engineering of the Drosophila genome with the phiC31 integrase system and will likely benefit research in Drosophila and other insects. PMID- 15126398 TI - Nucleotide variation and recombination along the fourth chromosome in Drosophila simulans. AB - The fourth chromosome of Drosophila melanogaster and its sister species are believed to be nonrecombining and have been a model system for testing predictions of the effects of selection on linked, neutral variation. We recently examined nucleotide variation along the chromosome of D. melanogaster and revealed that a low average level of recombination could be associated with considerably high levels of nucleotide variation. In this report, we further investigate the variation along the fourth chromosome of D. simulans. We sequenced 12 gene regions evenly distributed along the fourth chromosome for a worldwide collection of 11 isofemale lines and 5 gene regions in a local population of 10 isofemale lines from South America. In contrast to predictions for regions of very low recombination, these data reveal that the variation levels in many gene regions, including an intron region of the ci gene, vary considerably along the fourth chromosome. Nucleotide diversity ranged from 0.0010 to 0.0074 in 9 gene regions interspersed with several regions of greatly reduced variation. Tests of recombination indicate that the recombination level is not as low as previously thought, likely an order of magnitude higher than that in D. melanogaster. Finally, estimates of the recombination parameters are shown to support a crossover-plus-conversion model. PMID- 15126399 TI - Isolation and cytogenetic characterization of male meiotic mutants of Drosophila melanogaster. AB - Proper segregation of homologous chromosomes in meiosis I is ensured by pairing of homologs and maintenance of sister chromatid cohesion. In male Drosophila melanogaster, meiosis is achiasmatic and homologs pair at limited chromosome regions called pairing sites. We screened for male meiotic mutants to identify genes required for normal pairing and disjunction of homologs. Nondisjunction of the sex and the fourth chromosomes in male meiosis was scored as a mutant phenotype. We screened 2306 mutagenized and 226 natural population-derived second and third chromosomes and obtained seven mutants representing different loci on the second chromosome and one on the third. Five mutants showed relatively mild effects (<10% nondisjunction). mei(2)yh149 and mei(2)yoh7134 affected both the sex and the fourth chromosomes, mei(2)yh217 produced possible sex chromosome specific nondisjunction, and mei(2)yh15 and mei(2)yh137 produced fourth chromosome-specific nondisjunction. mei(2)yh137 was allelic to the teflon gene required for autosomal pairing. Three mutants exhibited severe defects, producing >10% nondisjunction of the sex and/or the fourth chromosomes. mei(2)ys91 (a new allele of the orientation disruptor gene) and mei(3)M20 induced precocious separation of sister chromatids as early as prometa-phase I. mei(2)yh92 predominantly induced nondisjunction at meiosis I that appeared to be the consequence of failure of the separation of paired homologous chromosomes. PMID- 15126401 TI - Drosophila male-specific lethal 2 protein controls sex-specific expression of the roX genes. AB - The MSL complex of Drosophila upregulates transcription of the male X chromosome, equalizing male and female X-linked gene expression. Five male-specific lethal proteins and at least one of the two noncoding roX RNAs are essential for this process. The roX RNAs are required for the localization of MSL complexes to the X chromosome. Although the mechanisms directing targeting remain speculative, the ratio of MSL protein to roX RNA influences localization of the complex. We examine the transcriptional regulation of the roX genes and show that MSL2 controls male-specific roX expression in the absence of any other MSL protein. We propose that this mechanism maintains a stable MSL/roX ratio that is favorable for localization of the complex to the X chromosome. PMID- 15126400 TI - Quantitative trait loci affecting starvation resistance in Drosophila melanogaster. AB - The ability to withstand periods of scarce food resources is an important fitness trait. Starvation resistance is a quantitative trait controlled by multiple interacting genes and exhibits considerable genetic variation in natural populations. This genetic variation could be maintained in the face of strong selection due to a trade-off in resource allocation between reproductive activity and individual survival. Knowledge of the genes affecting starvation tolerance and the subset of genes that affect variation in starvation resistance in natural populations would enable us to evaluate this hypothesis from a quantitative genetic perspective. We screened 933 co-isogenic P-element insertion lines to identify candidate genes affecting starvation tolerance. A total of 383 P-element insertions induced highly significant and often sex-specific mutational variance in starvation resistance. We also used deficiency complementation mapping followed by complementation to mutations to identify 12 genes contributing to variation in starvation resistance between two wild-type strains. The genes we identified are involved in oogenesis, metabolism, and feeding behaviors, indicating a possible link to reproduction and survival. However, we also found genes with cell fate specification and cell proliferation phenotypes, which implies that resource allocation during development and at the cellular level may also influence the phenotypic response to starvation. PMID- 15126402 TI - Molecular and functional analysis of scalloped recessive lethal alleles in Drosophila melanogaster. AB - The Drosophila melanogaster scalloped (sd) gene is a homolog of the human TEF-1 gene and is a member of the TEA/ATTS domain-containing family of transcription factors. In Drosophila, sd is involved in wing development as well as neural development. Herein, data are presented from a molecular analysis of five recessive lethal sd alleles. Only one of these alleles complements a viable allele associated with an sd mutant wing phenotype, suggesting that functions important for wing development are compromised by the noncomplementing alleles. Two of the wing noncomplementing alleles have mutations that help to define a VG binding domain for the SD protein in vivo, and another noncomplementing allele has a lesion within the TEA DNA-binding domain. The VG-binding domain overlaps with a domain important for viability of the fly, since two of the sd lethal lesions are located there. The fifth lethal affects a yet undefined motif lying just outside the VG-binding domain in the C-terminal direction that affects both wing phenotype and viability. This is the first example linking mutations affecting specific amino acids in the SD protein with phenotypic consequences for the organism. PMID- 15126403 TI - Nucleotide variation in the tinman and bagpipe homeobox genes of Drosophila melanogaster. AB - The tinman (tin) and bagpipe (bap) genes are members of the NK homeobox gene family of Drosophila, so that tin occupies a higher position than bap in the regulatory hierarchy. Little is known about the level and pattern of genetic polymorphism in homeobox genes. We have analyzed nucleotide polymorphism in 27 strains of Drosophila melanogaster and one each of D. simulans and D. sechellia, within two closely linked regions encompassing a partial sequence of tin and the complete sequence of bap. The two genes exhibit different levels and patterns of nucleotide diversity. Two sets of sharply divergent sequence types are detected for tin. The haplotype structure of bap is more complex: about half of the sequences are identical (or virtually so), while the rest are fairly heterogeneous. The level of silent nucleotide variability is 0.0063 for tin but significantly higher, 0.0141, for bap, a level of polymorphism comparable to the most polymorphic structural genes of D. melanogaster. Recombination rate and gene conversion are also higher for bap than for tin. There is strong linkage disequilibrium, with the highest values in the introns of both genes and exon II of bap. The patterns of polymorphism in tin and bap are not compatible with an equilibrium model of selective neutrality. We suggest that negative selection and demographic history are the major factors shaping the pattern of nucleotide polymorphism in the tin and bap genes; moreover, there are clear indications of positive selection in the bap gene. PMID- 15126404 TI - Global population genetic structure and male-mediated gene flow in the green sea turtle (Chelonia mydas): analysis of microsatellite loci. AB - We assessed the degree of population subdivision among global populations of green sea turtles, Chelonia mydas, using four microsatellite loci. Previously, a single-copy nuclear DNA study indicated significant male-mediated gene flow among populations alternately fixed for different mitochondrial DNA haplotypes and that genetic divergence between populations in the Atlantic and Pacific Oceans was more common than subdivisions among populations within ocean basins. Even so, overall levels of variation at single-copy loci were low and inferences were limited. Here, the markedly more variable microsatellite loci confirm the presence of male-mediated gene flow among populations within ocean basins. This analysis generally confirms the genetic divergence between the Atlantic and Pacific. As with the previous study, phylogenetic analyses of genetic distances based on the microsatellite loci indicate a close genetic relationship among eastern Atlantic and Indian Ocean populations. Unlike the single-copy study, however, the results here cannot be attributed to an artifact of general low variability and likely represent recent or ongoing migration between ocean basins. Sequence analyses of regions flanking the microsatellite repeat reveal considerable amounts of cryptic variation and homoplasy and significantly aid in our understanding of population connectivity. Assessment of the allele frequency distributions indicates that at least some of the loci may not be evolving by the stepwise mutation model. PMID- 15126405 TI - Mitochondrial cytochrome B DNA variation in the high-fecundity atlantic cod: trans-atlantic clines and shallow gene genealogy. AB - An analysis of sequence variation of 250 bp of the mitochondrial cytochrome b gene of 1278 Atlantic cod Gadus morhua ranging from Newfoundland to the Baltic shows four high-frequency (>8%) haplotypes and a number of rare and singleton haplotypes. Variation is primarily synonymous mutations. Natural selection acting directly on these variants is either absent or very weak. Common haplotypes show regular trans-Atlantic clines in frequencies and each of them reaches its highest frequency in a particular country. A shallow multifurcating constellation gene genealogy implies young age and recent turnover of polymorphism. Haplotypes characterizing populations at opposite ends of the geographic distribution in Newfoundland and the Baltic are mutationally closest together. The haplotypes are young and have risen rapidly in frequency. Observed differentiation among countries is due primarily to clinal variation. Hypotheses of historical isolation and polymorphisms balanced by local selection and gene flow are unlikely. Instead the results are explained by demic selection of mitochondria carried by highly fit females winning reproductive sweepstakes. By inference the Atlantic cod, a very high-fecundity vertebrate, is characterized by a high variance of offspring number and strong natural selection that leads to very low effective to actual population sizes. PMID- 15126406 TI - Evolution of DMY, a newly emergent male sex-determination gene of medaka fish. AB - The Japanese medaka fish Oryzias latipes has an XX/XY sex-determination system. The Y-linked sex-determination gene DMY is a duplicate of the autosomal gene DMRT1, which encodes a DM-domain-containing transcriptional factor. DMY appears to have originated recently within Oryzias, allowing a detailed evolutionary study of the initial steps that led to the new gene and new sex-determination system. Here I analyze the publicly available DMRT1 and DMY gene sequences of Oryzias species and report the following findings. First, the synonymous substitution rate in DMY is 1.73 times that in DMRT1, consistent with the male driven evolution hypothesis. Second, the ratio of the rate of nonsynonymous nucleotide substitution (d(N)) to that of synonymous substitution (d(S)) is significantly higher in DMY than in DMRT1. Third, in DMRT1, the d(N)/d(S) ratio for the DM domain is lower than that for non-DM regions, as expected from the functional importance of the DM domain. But in DMY, the opposite is observed and the DM domain is likely under positive Darwinian selection. Fourth, only one characteristic amino acid distinguishes all DMY sequences from all DMRT1 sequences, suggesting that a single amino acid change may be largely responsible for the establishment of DMY as the male sex-determination gene in medaka fish. PMID- 15126407 TI - Evolutionary relationships of major histocompatibility complex class I genes in simian primates. AB - New World monkeys (NWMs) occupy a critical phylogenetic position in elucidating the evolutionary process of major histocompatibility complex (MHC) class I genes in primates. From three subfamilies of Aotinae, Cebinae, and Atelinae, the 5' flanking regions of 18 class I genes are obtained and phylogenetically examined in terms of Alu/LINE insertion elements as well as the nucleotide substitutions. Two pairs of genes from Aotinae and Atelinae are clearly orthologous to human leukocyte antigen (HLA) -E and -F genes. Of the remaining 14 genes, 8 belong to the distinct group B, together with HLA-B and -C, to the exclusion of all other HLA class I genes. These NWM genes are classified into four groups, designated as NWM-B1, -B2, -B3, and -B4. Of these, NWM-B2 is orthologous to HLA-B/C. Also, orthologous relationships of NWM-B1, -B2, and -B3 exist among different families of Cebidae and Atelidae, which is in sharp contrast to the genus-specific gene organization within the subfamily Callitrichinae. The other six genes belong to the distinct group G. However, a clade of these NWM genes is almost equally related to HLA-A, -J, -G, and -K, and there is no evidence for their orthologous relationships to HLA-G. It is argued that class I genes in simian primates duplicated extensively in their common ancestral lineage and that subsequent evolution in descendant species has been facilitated mainly by independent loss of genes. PMID- 15126408 TI - Integration and modularity of quantitative trait locus effects on geometric shape in the mouse mandible. AB - The mouse mandible has long served as a model system for complex morphological structures. Here we use new methodology based on geometric morphometrics to test the hypothesis that the mandible consists of two main modules, the alveolar region and the ascending ramus, and that this modularity is reflected in the effects of quantitative trait loci (QTL). The shape of each mandible was analyzed by the positions of 16 morphological landmarks and these data were analyzed using Procrustes analysis. Interval mapping in the F(2) generation from intercrosses of the LG/J and SM/J strains revealed 33 QTL affecting mandible shape. The QTL effects corresponded to a variety of shape changes, but ordination or a parametric bootstrap test of clustering did not reveal any distinct groups of QTL that would affect primarily one module or the other. The correlations of landmark positions between the two modules tended to be lower than the correlations between arbitrary subsets of landmarks, indicating that the modules were relatively independent of each other and confirming the hypothesized location of the boundary between them. While these results are in agreement with the hypothesis of modularity, they also underscore that modularity is a question of the relative degrees to which QTL contribute to different traits, rather than a question of discrete sets of QTL contributing to discrete sets of traits. PMID- 15126409 TI - Integrating genetic linkage maps with pachytene chromosome structure in maize. AB - Genetic linkage maps reveal the order of markers based on the frequency of recombination between markers during meiosis. Because the rate of recombination varies along chromosomes, it has been difficult to relate linkage maps to chromosome structure. Here we use cytological maps of crossing over based on recombination nodules (RNs) to predict the physical position of genetic markers on each of the 10 chromosomes of maize. This is possible because (1). all 10 maize chromosomes can be individually identified from spreads of synaptonemal complexes, (2). each RN corresponds to one crossover, and (3). the frequency of RNs on defined chromosomal segments can be converted to centimorgan values. We tested our predictions for chromosome 9 using seven genetically mapped, single copy markers that were independently mapped on pachytene chromosomes using in situ hybridization. The correlation between predicted and observed locations was very strong (r(2) = 0.996), indicating a virtual 1:1 correspondence. Thus, this new, high-resolution, cytogenetic map enables one to predict the chromosomal location of any genetically mapped marker in maize with a high degree of accuracy. This novel approach can be applied to other organisms as well. PMID- 15126410 TI - Dynamic changes in the distribution of a satellite homologous to intergenic 26 18S rDNA spacer in the evolution of Nicotiana. AB - An approximately 135-bp sequence called the A1/A2 repeat was isolated from the transcribed region of the 26-18S rDNA intergenic spacer (IGS) of Nicotiana tomentosiformis. Fluorescence in situ hybridization (FISH) and Southern blot analysis revealed its occurrence as an independent satellite (termed an A1/A2 satellite) outside of rDNA loci in species of Nicotiana section Tomentosae. The chromosomal location, patterns of genomic dispersion, and copy numbers of its tandemly arranged units varied between the species. In more distantly related Nicotiana species the A1/A2 repeats were found only at the nucleolar organizer regions (NOR). There was a trend toward the elimination of the A1/A2 satellite in N. tabacum (tobacco), an allotetraploid with parents closely related to the diploids N. sylvestris and N. tomentosiformis. This process may have already commenced in an S(3) generation of synthetic tobacco. Cytosine residues in the IGS were significantly hypomethylated compared with the A1/A2 satellite. There was no clear separation between the IGS and satellite fractions in sequence analysis of individual clones and we found no evidence for CG suppression. Taken together the data indicate a dynamic nature of the A1/A2 repeats in Nicotiana genomes, with evidence for recurrent integration, copy number expansions, and contractions. PMID- 15126411 TI - Molecular organization of large fragments in the maize B chromosome: indication of a novel repeat. AB - The supernumerary B chromosome has no apparent effects on plant growth, and its molecular makeup is difficult to unravel, due to its high homology to the normal complement, which prevents conventional cloning. This difficulty was overcome previously by microdissecting the B chromosome under the microscope to result in 19 B clones, one of which is B specific and highly repetitive, dispersing over one-third of the B long arm and most regions of the centromeric knob. To gain insights into the molecular structure of the B chromosome, this sequence was used to screen a genomic library constructed from W22 carrying 16 B's. Five clones (>10 kb each) were isolated, and all were repetitive, showing homology with A chromosomes in Southern and FISH analyses. Two of them were further characterized and sequenced. Each is composed of several restriction fragments with variable degrees of repetitiveness. Some of these are B specific and others have variable degrees of homology with the A chromosomes. The order of each characteristic group is not contiguous; they intersperse within those of other groups. Sequence analysis reveals that their sequences ( approximately 26 kb) have no homology with any published gene other than sequences of transposable elements (retrotransposons and MITEs) and the B as well as the A centromeres. We uncovered a 1.6-kb CL-repeat sequence, seven units of which were present in the two clones in defective forms. Those repeats mostly arrange in tandem array in the B chromosome. Moreover, we detected transposition of a retrotransposon and a MITE element involved in the genesis of these two sequences. PMID- 15126412 TI - Sibship reconstruction from genetic data with typing errors. AB - Likelihood methods have been developed to partition individuals in a sample into full-sib and half-sib families using genetic marker data without parental information. They invariably make the critical assumption that marker data are free of genotyping errors and mutations and are thus completely reliable in inferring sibships. Unfortunately, however, this assumption is rarely tenable for virtually all kinds of genetic markers in practical use and, if violated, can severely bias sibship estimates as shown by simulations in this article. I propose a new likelihood method with simple and robust models of typing error incorporated into it. Simulations show that the new method can be used to infer full- and half-sibships accurately from marker data with a high error rate and to identify typing errors at each locus in each reconstructed sib family. The new method also improves previous ones by adopting a fresh iterative procedure for updating allele frequencies with reconstructed sibships taken into account, by allowing for the use of parental information, and by using efficient algorithms for calculating the likelihood function and searching for the maximum-likelihood configuration. It is tested extensively on simulated data with a varying number of marker loci, different rates of typing errors, and various sample sizes and family structures and applied to two empirical data sets to demonstrate its usefulness. PMID- 15126413 TI - Mapping quantitative trait loci in F2 incorporating phenotypes of F3 progeny. AB - In plants and laboratory animals, QTL mapping is commonly performed using F(2) or BC individuals derived from the cross of two inbred lines. Typical QTL mapping statistics assume that each F(2) individual is genotyped for the markers and phenotyped for the trait. For plant traits with low heritability, it has been suggested to use the average phenotypic values of F(3) progeny derived from selfing F(2) plants in place of the F(2) phenotype itself. All F(3) progeny derived from the same F(2) plant belong to the same F(2:3) family, denoted by F(2:3). If the size of each F(2:3) family (the number of F(3) progeny) is sufficiently large, the average value of the family will represent the genotypic value of the F(2) plant, and thus the power of QTL mapping may be significantly increased. The strategy of using F(2) marker genotypes and F(3) average phenotypes for QTL mapping in plants is quite similar to the daughter design of QTL mapping in dairy cattle. We study the fundamental principle of the plant version of the daughter design and develop a new statistical method to map QTL under this F(2:3) strategy. We also propose to combine both the F(2) phenotypes and the F(2:3) average phenotypes to further increase the power of QTL mapping. The statistical method developed in this study differs from published ones in that the new method fully takes advantage of the mixture distribution for F(2:3) families of heterozygous F(2) plants. Incorporation of this new information has significantly increased the statistical power of QTL detection relative to the classical F(2) design, even if only a single F(3) progeny is collected from each F(2:3) family. The mixture model is developed on the basis of a single-QTL model and implemented via the EM algorithm. Substantial computer simulation was conducted to demonstrate the improved efficiency of the mixture model. Extension of the mixture model to multiple QTL analysis is developed using a Bayesian approach. The computer program performing the Bayesian analysis of the simulated data is available to users for real data analysis. PMID- 15126414 TI - Genomic background predicts the fate of duplicated genes: evidence from the yeast genome. AB - Gene duplication with subsequent divergence plays a central role in the acquisition of genes with novel function and complexity during the course of evolution. With reduced functional constraints or through positive selection, these duplicated genes may experience accelerated evolution. Under the model of subfunctionalization, loss of subfunctions leads to complementary acceleration at sites with two copies, and the difference in average rate between the sequences may not be obvious. On the other hand, the classical model of neofunctionalization predicts that the evolutionary rate in one of the two duplicates is accelerated. However, the classical model does not tell which of the duplicates experiences the acceleration in evolutionary rate. Here, we present evidence from the Saccharomyces cerevisiae genome that a duplicate located in a genomic region with a low-recombination rate is likely to evolve faster than a duplicate in an area of high recombination. This observation is consistent with population genetics theory that predicts that purifying selection is less effective in genomic regions of low recombination (Hill-Robertson effect). Together with previous studies, our results suggest the genomic background (e.g., local recombination rate) as a potential force to drive the divergence between nontandemly duplicated genes. This implies the importance of structure and complexity of genomes in the diversification of organisms via gene duplications. PMID- 15126415 TI - Recovering frequencies of known haplotype blocks from single-nucleotide polymorphism allele frequencies. AB - Prospects for large-scale association studies rely on economical methods and powerful analysis. Representing available SNPs by small subsets and measuring allele frequencies on pooled DNA samples each improve genotyping cost effectiveness, while haplotype analysis may highlight associations in otherwise underpowered studies. This manuscript provides the mathematical framework to integrate these methodologies. PMID- 15126416 TI - Unexpected role of TNF-alpha in graft versus host reaction (GVHR): donor-derived TNF-alpha suppresses GVHR via inhibition of IFN-gamma-dependent donor type-1 immunity. AB - Graft versus host disease (GVHD) is a major complication of allogeneic hematopoietic stem cell transplantation, leading to significant morbidity and mortality. Host-derived TNF-alpha play a role in the induction of allo-reactive donor T cell activation and the pathogenesis of GVHD. On the other hand, the precise role of donor-derived TNF-alpha in GVHD remains unclear. To elucidate this issue, we designed an acute GVHD model using (B6 x D2) F1 recipient mice transferred with spleen cells derived from either wild-type or TNF-alpha(-/-) C57BL/6 mice. Surprisingly, we found that spleen cells from TNF-alpha(-/-) mice induce more severe graft versus host reaction (GVHR) than wild-type spleen cells upon transfer into B6D2F1 mice. Transplantation of TNF-alpha(-/-) mouse spleen cells was associated with enhanced anti-host CTL generation and augmented deletion of host cells. Moreover, mice receiving TNF-alpha(-/-) cells showed significantly higher levels of serum IFN-gamma, which was mainly produced by donor CD8+ T cells. We also demonstrated that TNF-alpha deficiency in donor spleen cells caused a marked elevation of TNF-alpha producing capacity by LPS stimulated host macrophages. Such enhanced GVHR was completely prevented by using TNF-alpha(-/-)IFN-gamma(-/-) splenic cells. Our findings demonstrate, for the first time, that donor-derived TNF-alpha suppress GVHR by inhibiting IFN-gamma dependent donor type-1 immunity which is essential for host TNF-alpha elevation. PMID- 15126417 TI - Duplicated copies of the bovine JH locus contribute to the Ig repertoire. AB - We report the cloning and analysis of a bovine JH locus comprising a DQ52 segment, six JH segments and sequence to a 5' H chain intronic enhancer. The contig was mapped to BTA 11 and evidence was found for rearrangement of the sixth JH segment at a low but detectable frequency. In contrast, the fourth segment present at a second copy of the bovine JH locus mapping to BTA 21 was found to rearrange at high-frequency, forming FR4 in the majority of bovine Ig H chains. The data thus show that bovine H chains can be generated from segments at two distinct genomic locations. Further investigation should establish if rearrangement takes place at each locus or if the participating segments are brought together from different chromosomal locations by less conventional processes (for example by gene conversion or trans-chromosomal rearrangement). PMID- 15126419 TI - Private efforts pick up stem cell slack. PMID- 15126418 TI - Expression profiles and functional implications of p53-like transcription factors in thymic epithelial cell subtypes. AB - In this study, we investigated the localization and functional significance of p53 tumor suppressor-like molecules, p63 and p73, in human thymic epithelial cells (TECs). Immunohistochemical studies showed particular distribution profiles of p63 and p73 in thymic epithelium, in which cortical TECs preferentially expressed p63 in their nuclei whereas subcapsular and medullary TECs expressed both p63 and p73 in their nuclei. The wide distribution of p63 in TECs was further suggested by studies using TECs of primary culture. In vitro studies using two human TEC lines demonstrated that p63 was capable of up-regulating intercellular adhesion molecule-1 (ICAM-1) and enhancing the production of IL-6 and IL-8. Moreover, in vitro studies also indicated that p73, but not p63, had the capacity to induce granulocyte macrophage colony stimulating factor (GM-CSF) and granulocyte colony stimulating factor (G-CSF) in the TEC lines. These findings suggest that p63 would regulate the cell adhesive property through ICAM 1/LFA-1 interaction and the production of IL-6 and IL-8, probably in all TEC subtypes. p73 in subcapslar and medullary TECs was suggested to play a role in the regulation of the production of GM-CSF and G-CSF, which might stimulate other stromal cells such as dendritic cells, macrophages and endothelial cells around these regions. PMID- 15126420 TI - Suicide caution stamped on antidepressants. PMID- 15126421 TI - Metabolic syndrome recasts old cardiac, diabetes risk factors as a "new" entity. PMID- 15126426 TI - Migraine as a risk factor for subclinical brain lesions. PMID- 15126427 TI - Effects of breast augmentation on accuracy of mammography. PMID- 15126428 TI - Safety of newer antiepileptic drugs. PMID- 15126429 TI - Aggressive behaviors and injuries among nursing home residents. PMID- 15126430 TI - Physicians as citizens. PMID- 15126431 TI - Physicians as citizens. PMID- 15126432 TI - Physicians as citizens. PMID- 15126433 TI - Surrogate consent for live organ donation. PMID- 15126434 TI - Hospital volume and quality. PMID- 15126435 TI - Preimplantation HLA testing. AB - CONTEXT: Preimplantation genetic diagnosis (PGD) has become an option for couples for whom termination of an affected pregnancy identified by traditional prenatal diagnosis is unacceptable and is applicable to indications beyond those of prenatal diagnosis, such as HLA matching to affected siblings to provide stem cell transplantation. OBJECTIVE: To describe preimplantation HLA typing, not involving identification of a causative gene, for couples who had children with bone marrow disorders at need for HLA-matched stem cell transplantation. DESIGN, SETTING, AND PARTICIPANTS: HLA matching procedures conducted at a single site during 2002-2003 in an in vitro fertilization program for 9 couples with children affected by acute lymphoid leukemia, acute myeloid leukemia, or Diamond-Blackfan anemia requiring HLA-matched stem cell transplantation. In 13 clinical cycles, DNA in single blastomeres removed from 8-cell embryos following in vitro fertilization was analyzed for HLA genes simultaneously with analysis for short tandem repeats in the HLA region to select and transfer only those embryos that were HLA matched to affected siblings. MAIN OUTCOME MEASURES: Results of HLA matching and pregnancy outcome. RESULTS: As a result of testing a total of 199 embryos, 45 (23%) HLA-matched embryos were selected, of which 28 were transferred in 12 clinical cycles, resulting in 5 singleton pregnancies and birth of 5 HLA matched healthy children. CONCLUSION: This is the first known experience of preimplantation HLA typing performed without PGD for a causative gene, providing couples with a realistic option of having HLA-matched offspring to serve as potential donors of stem cells for their affected siblings. PMID- 15126436 TI - Long-term efficacy of BCG vaccine in American Indians and Alaska Natives: A 60 year follow-up study. AB - CONTEXT: The duration of protection from tuberculosis of BCG vaccines is not known. OBJECTIVE: To determine the long-term duration of protection of a BCG vaccine that was previously found to be efficacious. DESIGN: Retrospective record review using Indian Health Service records, tuberculosis registries, death certificates, and supplemental interviews with trial participants. SETTING AND PARTICIPANTS: Follow-up for the period 1948-1998 among American Indians and Alaska Natives who participated in a placebo-controlled BCG vaccine trial during 1935-1938 and who were still at risk of developing tuberculosis. Data from 1483 participants in the BCG vaccine group and 1309 in the placebo group were analyzed. MAIN OUTCOME MEASURES: Efficacy of BCG vaccine, calculated for each 10 year interval using a Cox regression model with time-dependent variables based on tuberculosis events occurring after December 31, 1947 (end of prospective case finding). RESULTS: The overall incidence of tuberculosis was 66 and 138 cases per 100 000 person-years in the BCG vaccine and placebo groups, respectively, for an estimate of vaccine efficacy of 52% (95% confidence interval, 27%-69%). Adjustments for age at vaccination, tribe, subsequent BCG vaccination, chronic medical illness, isoniazid use, and bacille Calmette-Guerin strain did not substantially affect vaccine efficacy. There was slight but not statistically significant waning of the efficacy of BCG vaccination over time, greater among men than women. CONCLUSION: In this trial, BCG vaccine efficacy persisted for 50 to 60 years, suggesting that a single dose of an effective BCG vaccine can have a long duration of protection. PMID- 15126437 TI - Lipid-lowering therapy and in-hospital mortality following major noncardiac surgery. AB - CONTEXT: Cardiovascular complications following major noncardiac surgery are an important source of perioperative morbidity and mortality. Although lipid lowering medications are considered a key component in the primary and secondary prevention of cardiovascular disease, their potential benefit during the perioperative period is uncertain. OBJECTIVE: To examine the association between treatment with lipid-lowering medications and in-hospital mortality following major noncardiac surgery. DESIGN, SETTING, AND PATIENTS: A retrospective cohort study based on hospital discharge and pharmacy records of 780,591 patients aged 18 years or older who underwent major noncardiac surgery from January 1, 2000, to December 31, 2001, at any 1 of 329 hospitals throughout the United States. Only patients who survived through at least the second hospital day were included. Lipid-lowering therapy was defined as use during the first 2 hospital days. Propensity matching was used to adjust for numerous baseline differences. MAIN OUTCOME MEASURE: In-hospital mortality. RESULTS: Of the 780,591 patients, 77,082 patients (9.9%) received lipid-lowering therapy perioperatively and 23 100 (2.96%) died during the hospitalization. Treatment with lipid-lowering agents was associated with lower crude mortality (2.13% vs 3.05%, P<.001). In an analysis using matching by propensity score, 1595 patients (2.18%) treated with lipid lowering medications died compared with 4158 patients (3.15%) who did not receive therapy or in whom treatment was initiated after the second day (P<.001). After adjusting for residual differences in the propensity matched groups using conditional logistic regression, risk of mortality remained lower among treated patients (adjusted odds ratio [OR], 0.62; 95% confidence interval [CI], 0.58 0.67). Based on this adjusted OR, the number needed to treat to prevent a postoperative death in the propensity matched cohort was 85 (95% CI, 77-98) and varied from 186 among patients at lowest risk to 30 among those with a revised cardiac risk index score of 4 or more. In a further analysis using the entire study cohort and adjusting for quintile of propensity, a significant effect of treatment persisted (adjusted OR, 0.71; 95% CI, 0.67-0.75). CONCLUSIONS: Treatment with lipid-lowering agents may reduce risk of death following major noncardiac surgery. Clinical trials are required to confirm this observation. PMID- 15126438 TI - Treatment for cervical intraepithelial neoplasia and risk of preterm delivery. AB - CONTEXT: It is unclear whether treatments for cervical intraepithelial neoplasia (CIN) increase the subsequent risk of preterm delivery. Most studies have lacked sufficient sample size, mixed heterogeneous subtypes of preterm delivery, and failed to control for confounding factors. OBJECTIVE: To determine whether cervical laser and loop electrosurgical excision procedure (LEEP) treatments increase risk of preterm delivery and its subtypes. DESIGN, SETTING, AND PARTICIPANTS: Retrospective cohort study conducted among women evaluated at a colposcopy clinic serving Auckland, New Zealand (1988-2000), comparing delivery outcomes of untreated women (n = 426) and those treated (n = 652) with laser conization, laser ablation, or LEEP. Record linkage using unique health identifiers identified women who had subsequent deliveries. MAIN OUTCOME MEASURES: Total preterm delivery and its subtypes, spontaneous labor and premature rupture of membranes before 37 weeks' gestation (pPROM). RESULTS: The overall rate of preterm delivery was 13.8%. The rate of pPROM was 6.2% and the rate of spontaneous preterm delivery was 3.8%. Analyses showed no significant increase in risk of total preterm delivery (adjusted relative risk [aRR], 1.1; 95% confidence interval [CI], 0.8-1.5) or spontaneous preterm delivery (aRR, 1.3; 95% CI, 0.7-2.6) for any treatment. Risk of pPROM was significantly increased following treatment with laser conization (aRR, 2.7; 95% CI, 1.3-5.6) or LEEP (aRR, 1.9; 95% CI, 1.0-3.8), but not laser ablation (aRR, 1.1; 95% CI, 0.5-2.4). Moreover, risk of pPROM and total preterm delivery increased significantly with increasing height of tissue removed from the cervix in conization. Women in the highest tertile of cone height (> or =1.7 cm) had a greater than 3-fold increase in risk of pPROM compared with untreated women (aRR, 3.6; 95% CI, 1.8-7.5). CONCLUSIONS: LEEP and laser cone treatments were associated with significantly increased risk of pPROM. Careful consideration should be given to treatment of CIN in women of reproductive age, especially when treatment might reasonably be delayed or targeted to high-risk cases. PMID- 15126439 TI - Trends in blood pressure among children and adolescents. AB - CONTEXT: The prevalence of overweight among children and adolescents increased between 1988 and 2000. The change in blood pressure among children and adolescents over that time and the role of overweight is unknown. OBJECTIVE: To examine trends in systolic and diastolic blood pressure among children and adolescents between 1988 and 2000. DESIGN, SETTING, AND POPULATION: Two serially conducted cross-sectional studies using nationally representative samples of children and adolescents, aged 8 to 17 years, from the third National Health and Nutrition Examination Survey (NHANES III) conducted in 1988-1994 (n = 3496) and NHANES 1999-2000 (n = 2086). MAIN OUTCOME MEASURES: Systolic and diastolic blood pressure levels. RESULTS: In 1999-2000, the mean (SE) systolic blood pressure was 106.0 (0.3) mm Hg and diastolic blood pressure was 61.7 (0.5) mm Hg. After adjustment for age, mean systolic blood pressure was 1.6 mm Hg higher among non Hispanic black girls (P =.11) and 2.9 mm Hg higher among non-Hispanic black boys (P<.001) compared with non-Hispanic whites. Among Mexican Americans, girls' systolic blood pressure was 1.0 mm Hg higher (P =.21) and boys' was 2.7 mm Hg higher (P<.001) compared with non-Hispanic whites (P<.001). With further adjustment for body mass index, these differences were attenuated. After age, race/ethnicity, and sex standardization, systolic blood pressure was 1.4 (95% confidence interval [CI], 0.6-2.2) mm Hg higher (P<.001) and diastolic blood pressure was 3.3 (95% CI, 2.1-4.5) mm Hg higher in 1999-2000 (P<.001) compared with 1988-1994. With further adjustment for differences in the body mass index distribution in 1988-1994 and 1999-2000, the increase in systolic blood pressure was reduced by 29% and diastolic blood pressure was reduced by 12%. CONCLUSIONS: Blood pressure has increased over the past decade among children and adolescents. This increase is partially attributable to an increased prevalence of overweight. PMID- 15126440 TI - Prevalence of marijuana use disorders in the United States: 1991-1992 and 2001 2002. AB - CONTEXT: Among illicit substance use disorders, marijuana use disorders are the most prevalent in the population. Yet, information about the prevalence of current Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) marijuana use disorders and how prevalence has changed is lacking. OBJECTIVE: To examine changes in the prevalence of marijuana use, abuse, and dependence in the United States between 1991-1992 and 2001-2002. DESIGN, SETTING, AND PARTICIPANTS: Face-to-face interviews were conducted in 2 large national surveys conducted 10 years apart: the 1991-1992 National Longitudinal Alcohol Epidemiologic Survey ([NLAES] n = 42,862) and the 2001-2002 National Epidemiologic Survey on Alcohol and Related Conditions ([NESARC] n = 43,093). MAIN OUTCOME MEASURES: Rates of past year marijuana use, abuse, and dependence. RESULTS: Among the adult US population, the prevalence of marijuana use remained stable at about 4.0% over the past decade. In contrast, the prevalence of DSM-IV marijuana abuse or dependence significantly (P =.01) increased between 1991-1992 (1.2%) and 2001-2002 (1.5%), with the greatest increases observed among young black men and women (P<.001) and young Hispanic men (P =.006). Further, marijuana use disorders among marijuana users significantly increased (P =.002) in the absence of increased frequency and quantity of marijuana use, suggesting that the concomitant increase in potency of delta-9-tetrahydrocannabinol (Delta9-THC) may have contributed to the rising rates. CONCLUSIONS: Despite the stability in the overall prevalence of marijuana use, more adults in the United States had a marijuana use disorder in 2001-2002 than in 1991-1992. Increases in the prevalence of marijuana use disorders were most notable among young black men and women and young Hispanic men. Although rates of marijuana abuse and dependence did not increase among young white men and women, their rates have remained high. The results of this study underscore the need to develop and implement new prevention and intervention programs targeted at youth, particularly minority youth. PMID- 15126441 TI - External fixation in orthopedics. PMID- 15126442 TI - Conception for donation. PMID- 15126443 TI - A booster for tuberculosis vaccines. PMID- 15126444 TI - STUDENTJAMA. The evolution and evaluation of modern medical education. PMID- 15126445 TI - STUDENTJAMA. The Flexner report and the standardization of American medical education. PMID- 15126446 TI - STUDENTJAMA. Medical education in modern China. PMID- 15126447 TI - STUDENTJAMA. Cultural competence training in US medical schools. PMID- 15126448 TI - STUDENTJAMA. Evaluating competence in medical students. PMID- 15126449 TI - JAMA patient page. Bone fractures. PMID- 15126450 TI - OmpR and LeuO positively regulate the Salmonella enterica serovar Typhi ompS2 porin gene. AB - The Salmonella enterica serovar Typhi ompS2 gene codes for a 362-amino-acid outer membrane protein that contains motifs common to the porin superfamily. It is expressed at very low levels compared to the major OmpC and OmpF porins, as observed for S. enterica serovar Typhi OmpS1, Escherichia coli OmpN, and Klebsiella pneumoniae OmpK37 quiescent porins. A region of 316 bp, between nucleotides -413 and -97 upstream of the transcriptional start point, is involved in negative regulation, as its removal resulted in a 10-fold increase in ompS2 expression in an S. enterica serovar Typhi wild-type strain. This enhancement in expression was not observed in isogenic mutant strains, which had specific deletions of the regulatory ompB (ompR envZ) operon. Furthermore, ompS2 expression was substantially reduced in the presence of the OmpR D55A mutant, altered in the major phosphorylation site. Upon random mutagenesis, a mutant where the transposon had inserted into the upstream regulatory region of the gene coding for the LeuO regulator, showed an increased level of ompS2 expression. Augmented expression of ompS2 was also obtained upon addition of cloned leuO to the wild-type strain, but not in an ompR isogenic derivative, consistent with the notion that the transposon insertion had increased the cellular levels of LeuO and with the observed dependence on OmpR. Moreover, LeuO and OmpR bound in close proximity, but independently, to the 5' upstream regulatory region. Thus, the OmpR and LeuO regulators positively regulate ompS2. PMID- 15126451 TI - TtgV bound to a complex operator site represses transcription of the promoter for the multidrug and solvent extrusion TtgGHI pump. AB - The TtgGHI efflux pump of Pseudomonas putida extrudes a variety of antibiotics and solvents. We show that the ttgGHI operon is transcribed in vitro and in vivo from a single promoter and not from two overlapping promoters as previously proposed. The expression of this promoter is controlled by the TtgV repressor, whose operator expands through four helical turns that overlap the -10 region of the promoter. We also show that TtgV is released from its operator on binding of effectors such as aliphatic alcohols. Mutational analysis of the ttgGHI promoter revealed that substitutions at -13, -12, and -8 yielded promoters that were unable to drive transcription whereas certain mutations at -9, -11, and -6 to -3 increased expression in vivo. The cause of the increased expression was either a decrease in the affinity of the TtgV protein for its operator or an increase in the affinity of RNA polymerase for the mutant promoters. PMID- 15126452 TI - Mutations in Haemophilus influenzae mismatch repair genes increase mutation rates of dinucleotide repeat tracts but not dinucleotide repeat-driven pilin phase variation rates. AB - High-frequency, reversible switches in expression of surface antigens, referred to as phase variation (PV), are characteristic of Haemophilus influenzae. PV enables this bacterial species, an obligate commensal and pathogen of the human upper respiratory tract, to adapt to changes in the host environment. Phase variable hemagglutinating pili are expressed by many H. influenzae isolates. PV involves alterations in the number of 5' TA repeats located between the -10 and 35 promoter elements of the overlapping, divergently orientated promoters of hifA and hifBCDE, whose products mediate biosynthesis and assembly of pili. Dinucleotide repeat tracts are destabilized by mismatch repair (MMR) mutations in Escherichia coli. The influence of mutations in MMR genes of H. influenzae strain Rd on dinucleotide repeat-mediated PV rates was investigated by using reporter constructs containing 20 5' AT repeats. Mutations in mutS, mutL, and mutH elevated rates approximately 30-fold, while rates in dam and uvrD mutants were increased 14- and 3-fold, respectively. PV rates of constructs containing 10 to 12 5' AT repeats were significantly elevated in mutS mutants of H. influenzae strains Rd and Eagan. An intact hif locus was found in 14 and 12% of representative nontypeable H. influenzae isolates associated with either otitis media or carriage, respectively. Nine or more tandem 5' TA repeats were present in the promoter region. Surprisingly, inactivation of mutS in two serotype b H. influenzae strains did not alter pilin PV rates. Thus, although functionally analogous to the E. coli MMR pathway and active on dinucleotide repeat tracts, defects in H. influenzae MMR do not affect 5' TA-mediated pilin PV. PMID- 15126453 TI - Positive control of swarming, rhamnolipid synthesis, and lipase production by the posttranscriptional RsmA/RsmZ system in Pseudomonas aeruginosa PAO1. AB - In Pseudomonas aeruginosa, the small RNA-binding, regulatory protein RsmA is a negative control element in the formation of several extracellular products (e.g., pyocyanin, hydrogen cyanide, PA-IL lectin) as well as in the production of N-acylhomoserine lactone quorum-sensing signal molecules. RsmA was found to control positively the ability to swarm and to produce extracellular rhamnolipids and lipase, i.e., functions contributing to niche colonization by P. aeruginosa. An rsmA null mutant was entirely devoid of swarming but produced detectable amounts of rhamnolipids, suggesting that factors in addition to rhamnolipids influence the swarming ability of P. aeruginosa. A small regulatory RNA, rsmZ, which antagonized the effects of RsmA, was identified in P. aeruginosa. Expression of the rsmZ gene was dependent on both the global regulator GacA and RsmA, increased with cell density, and was subject to negative autoregulation. Overexpression of rsmZ and a null mutation in rsmA resulted in quantitatively similar, negative or positive effects on target genes, in agreement with a model that postulates titration of RsmA protein by RsmZ RNA. PMID- 15126454 TI - Functional dissection of the XpsN (GspC) protein of the Xanthomonas campestris pv. campestris type II secretion machinery. AB - Type II secretion machinery is composed of 12 to 15 proteins for translocating extracellular proteins across the outer membrane. XpsL, XpsM, and XpsN are components of such machinery in the plant pathogen Xanthomonas campestris pv. campestris. All are bitopic cytoplasmic-membrane proteins, each with a large C terminal periplasmic domain. They have been demonstrated to form a dissociable ternary complex. By analyzing the C-terminally truncated XpsN and PhoA fusions, we discovered that truncation of the C-terminal 103 residues produced a functional protein, albeit present below detectable levels. Furthermore, just the first 46 residues, encompassing the membrane-spanning sequence (residues 10 to 32), are sufficient to keep XpsL and XpsM at normal abundance. XpsN46(His6), synthesized in Escherichia coli, is able to associate in a membrane-mixing experiment with the XpsL-XpsM complex preassembled in X. campestris pv. campestris. The XpsN N-terminal 46 residues are apparently sufficient not only for maintaining XpsL and XpsM at normal levels but also for their stable association. The membrane-spanning sequence of XpsN was not replaceable by that of TetA. However, coimmunoprecipitation with XpsL and XpsM was observed for XpsN97::PhoA, but not XpsN46::PhoA. Only XpsN97::PhoA is dominant negative. Single alanine substitutions for three charged residues within the region between residues 47 and 97 made the protein nonfunctional. In addition, the R78A mutant XpsN protein was pulled down by XpsL-XpsM(His6) immobilized on an Ni nitrilotriacetic acid column to a lesser extent than the wild-type XpsN. Therefore, in addition to the N-terminal 46 residues, the region between residues 47 and 97 of XpsN probably also plays an important role in interaction with XpsL XpsM. PMID- 15126455 TI - Dual control of Helicobacter pylori heat shock gene transcription by HspR and HrcA. AB - The HspR repressor regulates transcription of the groESL, hrcA-grpE-dnaK, and cbpA-hspR-orf operons of Helicobacter pylori. Here we show that two of the HspR regulated operons, namely, the groESL and dnaK operons, encoding the major cellular chaperone machineries are also regulated by the H. pylori homologue of the HrcA repressor. Similarly to the hspR mutation, deletion of the hrcA gene also leads to complete derepression of the Pgro and Phrc promoters. The presence of both HspR and HrcA is therefore necessary for regulated transcription from these promoters. HrcA binds directly to Pgro and Phrc, likely contacting two inverted repeats with similarity to the CIRCE motif, which are present on both promoters. HrcA regulation is, however, shown to depend on binding of the HspR protein, since deletion of the HspR-binding site of the Pgro promoter leads to loss of heat inducibility of this promoter. In contrast, transcription from the Pcbp promoter is regulated solely by HspR. HspR is also shown to form oligomers in vivo through a stretch of hydrophobic repeats between amino acid positions 66 and 97. The implications of these findings for the elucidation of the networks regulating heat shock gene expression in H. pylori are discussed. PMID- 15126456 TI - TcaR, a putative MarR-like regulator of sarS expression. AB - TcaR, which shares sequence homology with MarR-like transcriptional regulators, has been identified as a novel Staphylococcus aureus regulator affecting the expression of the global regulatory element SarS (SarH1), as well as that of the cell surface-associated protein SasF (N315-SA2439). Microarray analysis, confirmatory Northern blots, and genetic complementation experiments showed that TcaR upregulates sarS and thus spa transcription. In addition, it attenuates whole-length transcription of sasF, thereby producing a truncated transcript lacking the 3' terminus, which codes for the cell wall anchor motif. Hence, in strains containing an intact tcaR gene, TcaR is likely to decrease the amount of the surface-associated protein SasF and to increase that of the surface associated protein A. The widely used laboratory strains derived from NCTC8325 were found to be natural, truncated mutants of tcaR, harboring an inactive TcaR and therefore expressing very low levels of sarS. The data presented here identified TcaR as a further activator of sarS, and a modulator of sasF expression that has to be taken into account in studies of virulence gene expression in S. aureus. PMID- 15126457 TI - Assembly of the MexAB-OprM multidrug efflux system of Pseudomonas aeruginosa: identification and characterization of mutations in mexA compromising MexA multimerization and interaction with MexB. AB - The membrane fusion protein (MFP) component, MexA, of the MexAB-OprM multidrug efflux system of P. aeruginosa is proposed to link the inner (MexB) and outer (OprM) membrane components of this pump as a probable oligomer. A cross-linking approach confirmed the in vivo interaction of MexA and MexB, while a LexA-based assay for assessing protein-protein interaction similarly confirmed MexA multimerization. Mutations compromising the MexA contribution to antibiotic resistance but yielding wild-type levels of MexA were recovered and shown to map to two distinct regions within the N- and C-terminal halves of the protein. Most of the N-terminal mutations occurred at residues that are highly conserved in the MFP family (P68, G72, L91, A108, L110, and V129), consistent with these playing roles in a common feature of these proteins (e.g., oligomerization). In contrast, the majority of the C-terminal mutations occurred at residues poorly conserved in the MFP family (V264, N270, H279, V286, and G297), with many mapping to a region of MexA that corresponds to a region in the related MFP of Escherichia coli, AcrA, that is implicated in binding to its RND component, AcrB (C. A. Elkins and H. Nikaido, J. Bacteriol. 185:5349-5356, 2003). Given the noted specificity of MFP-RND interaction in this family of pumps, residues unique to MexA may well be important for and define the MexA interaction with its RND component, MexB. Still, all but one of the MexA mutations studied compromised MexA-MexB association, suggesting that native structure and/or proper assembly of the protein may be necessary for this. PMID- 15126458 TI - Effects of a gerF (lgt) mutation on the germination of spores of Bacillus subtilis. AB - One of the proteins of the membrane-bound receptors that recognize individual nutrients that trigger germination of spores of Bacillus subtilis contains the recognition sequence for diacylglycerol addition to a cysteine residue near the protein's N terminus. B. subtilis spores lacking the gerF (lgt) gene that codes for prelipoprotein diacylglycerol transferase exhibited significantly slowed germination in response to nutrient germinants as found previously, but germination of gerF spores with a mixture of Ca2+ and dipicolinic acid or with dodecylamine was normal, as was the spontaneous germination of gerF spores lacking all nutrient germinant receptors. The deleterious effects of the gerF mutation on nutrient germination were highest on germination triggered by the GerA nutrient receptor and were less so (but still significant) on germination triggered by the GerB nutrient receptor. However, there was little, if any, effect on GerK nutrient receptor-mediated spore germination. As predicted from the latter results, replacement by alanine of the cysteine residue to which diacylglycerol is thought to be added to these nutrient receptors had a large effect on GerA receptor function, less effect on GerB receptor function, and little, if any, effect on GerK receptor function. PMID- 15126459 TI - Drastic differences in Crh and HPr synthesis levels reflect their different impacts on catabolite repression in Bacillus subtilis. AB - In Bacillus subtilis, carbon catabolite repression (CCR) of catabolic genes is mediated by ATP-dependent phosphorylation of HPr and Crh. Here we show that the different efficiencies with which these two proteins contribute to CCR may be due to the drastic differences in their synthesis rates under conditions that cause CCR. PMID- 15126460 TI - Identification of rhtX and fptX, novel genes encoding proteins that show homology and function in the utilization of the siderophores rhizobactin 1021 by Sinorhizobium meliloti and pyochelin by Pseudomonas aeruginosa, respectively. AB - Rhizobactin 1021 is a hydroxymate siderophore produced by the soil bacterium Sinorhizobium meliloti 2011. A regulon comprising rhtA, encoding the outer membrane receptor protein for the ferrisiderophore; the biosynthesis operon rhbABCDEF; and rhrA, the Ara-C-like regulator of the receptor and biosynthesis genes has been previously described. We report the discovery of a gene, located upstream of rhbA and named rhtX (for "rhizobactin transport"), which is required, in addition to rhtA, to confer the ability to utilize rhizobactin 1021 on a strain of S. meliloti that does not naturally utilize the siderophore. Rhizobactin 1021 is structurally similar to aerobactin, which is transported in Escherichia coli via the IutA outer membrane receptor and the FhuCDB inner membrane transport system. E. coli expressing iutA and fhuCDB was found to also transport rhizobactin 1021. We demonstrated that RhtX alone could substitute for FhuCDB to transport rhizobactin 1021 in E. coli. RhtX shows similarity to a number of uncharacterized proteins which are encoded proximal to genes that are either known to be or predicted to be involved in iron acquisition. Among these is PA4218 of Pseudomonas aeruginosa, which is located close to the gene cluster that functions in pyochelin biosynthesis and outer membrane transport. PA4218 was mutated by allelic replacement, and the mutant was found to have a pyochelin utilization-defective phenotype. It is proposed that PA4218 be named fptX (for "ferripyochelin transport"). RhtX and FptX appear to be members of a novel family of permeases that function as single-subunit transporters of siderophores. PMID- 15126461 TI - Signal transduction cascade between EvgA/EvgS and PhoP/PhoQ two-component systems of Escherichia coli. AB - Transcriptional analysis of a constitutively active mutant of the EvgA/EvgS two component system of Escherichia coli resulted in enhanced expression of 13 PhoP/PhoQ-regulated genes, crcA, hemL, mgtA, ompT, phoP, phoQ, proP, rstA, rstB, slyB, ybjG, yrbL, and mgrB. This regulatory network between the two systems also occurred as a result of overproduction of the EvgA regulator; however, enhanced transcription of the phoPQ genes did not further activate expression of the PhoP/PhoQ-regulated genes. These results demonstrated signal transduction from the EvgA/EvgS system to the PhoP/PhoQ system in E. coli and also identified the genes that required the two systems for enhanced expression. This is one example of the intricate signal transduction networks that are posited to exist in E. coli. PMID- 15126462 TI - PhaQ, a new class of poly-beta-hydroxybutyrate (phb)-responsive repressor, regulates phaQ and phaP (phasin) expression in Bacillus megaterium through interaction with PHB. AB - Bacillus megaterium can produce poly-beta-hydroxybutyrate (PHB) as carbon and energy storage materials. We now report that the phaQ gene, which is located upstream of the phasin-encoding phaP gene, codes for a new class of transcriptional regulator that negatively controls expression of both phaQ and phaP. A PhaQ binding site that plays a role in this control has been identified by gel mobility shift assays and DNase I footprinting analysis. We have also provided evidence that PhaQ could sense the presence of PHB in vivo and that artificial PHB granules could inhibit the formation of PhaQ-DNA complex in vitro by binding to PhaQ directly. These suggest that PhaQ is a PHB-responsive repressor. PMID- 15126463 TI - Identification of an uptake hydrogenase required for hydrogen-dependent reduction of Fe(III) and other electron acceptors by Geobacter sulfurreducens. AB - Geobacter sulfurreducens, a representative of the family Geobacteraceae that predominates in Fe(III)-reducing subsurface environments, can grow by coupling the oxidation of hydrogen to the reduction of a variety of electron acceptors, including Fe(III), fumarate, and quinones. An examination of the G. sulfurreducens genome revealed two operons, hya and hyb, which appeared to encode periplasmically oriented respiratory uptake hydrogenases. In order to assess the roles of these two enzymes in hydrogen-dependent growth, Hya- and Hyb-deficient mutants were generated by gene replacement. Hyb was found to be required for hydrogen-dependent reduction of Fe(III), anthraquinone-2,6-disulfonate, and fumarate by resting cell suspensions and to be essential for growth with hydrogen and these three electron acceptors. Hya, in contrast, was not. These findings suggest that Hyb is an essential respiratory hydrogenase in G. sulfurreducens. PMID- 15126464 TI - Transcription of the Staphylococcus aureus cid and lrg murein hydrolase regulators is affected by sigma factor B. AB - The Staphylococcus aureus lrg and cid loci are homologous operons that have been shown to regulate murein hydrolase activity and affect sensitivity to penicillin. Although the mode of action of these operons has not been demonstrated, a model based on the similarities of the lrgA and cidA gene products to the bacteriophage holin family of proteins has been proposed. In this study, the transcription organization and regulation of these operons were examined by Northern blot analyses. Unexpectedly, cidB and a gene located immediately downstream, designated cidC, were found to be cotranscribed on a 2.7-kb transcript. Maximal cidBC transcription occurred during early exponential growth, and high-level transcription of cidBC was dependent on the rsbU-mediated activation of the alternative sigma factor B (sigmaB). In contrast, lrgAB transcription in stationary phase was negatively regulated by sigmaB. Although cidABC transcription was not detected by Northern blot analysis, reverse transcriptase PCR revealed that these genes are also cotranscribed as a single RNA message in early exponential growth. Primer extension analysis revealed the presence of two cidBC transcription start sites, but no apparent sigmaB-dependent promoter consensus sequence was identified in these regions. The rsbU gene was also shown to have a positive impact on murein hydrolase activity but a negligible effect on sensitivity to penicillin-induced killing. These results suggest that the lrgAB and cidBC genes may be part of the S. aureus sigmaB-controlled stress regulon. PMID- 15126465 TI - Expression of magA in Legionella pneumophila Philadelphia-1 is developmentally regulated and a marker of formation of mature intracellular forms. AB - Legionella pneumophila displays a biphasic developmental cycle in which replicating forms (RFs) differentiate postexponentially into highly infectious, cyst-like mature intracellular forms (MIFs). Using comparative protein profile analyses (MIFs versus RFs), we identified a 20-kDa protein, previously annotated as "Mip-like" protein, that was enriched in MIFs. However, this 20-kDa protein shared no similarity with Mip, a well-characterized peptidyl-prolyl isomerase of L. pneumophila, and for clarity we renamed it MagA (for "MIF-associated gene"). We monitored MagA levels across the growth cycle (in vitro and in vivo) by immunoblotting and established that MagA levels increased postexponentially in vitro (approximately 3-fold) and nearly 10-fold during MIF morphogenesis in HeLa cells. DNA sequence analysis of the magA locus revealed an upstream divergently transcribed gene, msrA, encoding a peptide methionine sulfoxide reductase and a shared promoter region containing direct and indirect repeat sequences as well as -10 hexamers often associated with stationary-phase regulation. While MagA has no known function, it contains a conserved CXXC motif commonly found in members of the thioredoxin reductase family and in AhpD reductases that are associated with alkylhydroperoxide reductase (AhpC), suggesting a possible role in protection from oxidative stress. MIFs from L. pneumophila strain Lp02 containing a magA deletion exhibited differences in Gimenez staining, as well as an apparent increase in cytopathology to HeLa cells, but otherwise were unaltered in virulence traits. As demonstrated by this study, MagA appears to be a MIF specific protein expressed late in intracellular growth that may serve as a useful marker of development. PMID- 15126466 TI - Temperature sensitivity caused by mutant release factor 1 is suppressed by mutations that affect 16S rRNA maturation. AB - To study the effect of slow termination on the protein synthesizing machinery, we isolated suppressors to a temperature-sensitive release factor 1 (RF1). Of 26 independent clones, five complementation groups have been identified, two of which are presented here. The first mutation disrupts a base pair in the transcription terminator stem for the rplM-rpsI operon, which encodes ribosomal proteins L13 and S9. We have found that this leads to readthrough of the terminator and that lower levels of transcript (compared to the results seen with the wild type) are found in the cell. This probably leads to decreased expression of the two proteins. The second mutation is a small deletion of the yrdC open reading frame start site, and it is not likely that the protein is expressed. Both mutant strains show an increased accumulation of 17S rRNA (immature 16S rRNA). Maturation of 16S rRNA is dependent on proper assembly of the ribosomal proteins, a process that is disturbed when proteins are missing. The function of the YrdC protein is not known, but it is able to bind to double-stranded RNA; therefore, we suggest that it is an assembly factor important for 30S subunit biogenesis. On the basis of our findings, we propose that lesser amounts of S9 or a lack of YrdC causes the maturation defect. We have shown that as a consequence of the maturation defect, fewer 70S ribosomes and polysomes are formed. This and other results suggest that it is the lowered concentration of functional ribosomes that suppresses the temperature sensitivity caused by the mutant RF1. PMID- 15126467 TI - Bacillus subtilis SalA (YbaL) negatively regulates expression of scoC, which encodes the repressor for the alkaline exoprotease gene, aprE. AB - During the course of screening for exoprotease-deficient mutants among Bacillus subtilis gene disruptants, a strain showing such a phenotype was identified. The locus responsible for this phenotype was the previously unknown gene ybaL, which we renamed salA. The predicted gene product encoded by salA belongs to the Mrp family, which is widely conserved among archaea, prokaryotes, and eukaryotes. Disruption of salA resulted in a decrease in the expression of a lacZ fusion of the aprE gene encoding the major extracellular alkaline protease. The decrease was recovered by the cloned salA gene on a plasmid, demonstrating that the gene is involved in aprE expression. Determination of the cis-acting region of SalA on the upstream region of aprE, together with epistatic analyses with scoC, abrB, and spo0A mutations that also affect aprE expression, suggested that salA deficiency affects aprE-lacZ expression through the negative regulator ScoC. Northern and reverse transcription-PCR analyses revealed enhanced levels of scoC transcripts in the salA mutant cells in the transition and early stationary phases. Concomitant with these observations, larger amounts of the ScoC protein were detected in the mutant cells by Western analysis. From these results we conclude that SalA negatively regulates scoC expression. It was also found that the expression of a salA-lacZ fusion was increased by salA deficiency, suggesting that salA is autoregulated. PMID- 15126468 TI - Agrobacterium rhizogenes GALLS protein substitutes for Agrobacterium tumefaciens single-stranded DNA-binding protein VirE2. AB - Agrobacterium tumefaciens and Agrobacterium rhizogenes transfer plasmid-encoded genes and virulence (Vir) proteins into plant cells. The transferred DNA (T-DNA) is stably inherited and expressed in plant cells, causing crown gall or hairy root disease. DNA transfer from A. tumefaciens into plant cells resembles plasmid conjugation; single-stranded DNA (ssDNA) is exported from the bacteria via a type IV secretion system comprised of VirB1 through VirB11 and VirD4. Bacteria also secrete certain Vir proteins into plant cells via this pore. One of these, VirE2, is an ssDNA-binding protein crucial for efficient T-DNA transfer and integration. VirE2 binds incoming ssT-DNA and helps target it into the nucleus. Some strains of A. rhizogenes lack VirE2, but they still transfer T-DNA efficiently. We isolated a novel gene from A. rhizogenes that restored pathogenicity to virE2 mutant A. tumefaciens. The GALLS gene was essential for pathogenicity of A. rhizogenes. Unlike VirE2, GALLS contains a nucleoside triphosphate binding motif similar to one in TraA, a strand transferase conjugation protein. Despite their lack of similarity, GALLS substituted for VirE2. PMID- 15126469 TI - Two separate quorum-sensing systems upregulate transcription of the same ABC transporter in Streptococcus pneumoniae. AB - Streptococcus pneumoniae secretes two different peptide pheromones used for intercellular communication. These peptides, which have completely unrelated primary structures, activate two separate signal transduction pathways, ComABCDE and BlpABCSRH, which regulate natural genetic transformation and bacteriocin production, respectively. Each signal transduction pathway contains a response regulator (ComE and BlpR, respectively) that activates transcription of target genes by binding to similar, but not identical, imperfect direct repeat motifs. In general the direct repeat binding sites are specific for one or the other of the two response regulators, ensuring that competence development and bacteriocin production are regulated separately. However, in the present study we show that the rate of transcription of an operon, encoding an ABC transporter of unknown function, can be stimulated by both peptide pheromones. We also show that this cross-induction is due to a hybrid direct repeat motif that can respond to both ComE and BlpR. To our knowledge this kind of convergent gene regulation by two separate two-component regulatory systems has not been described before in bacteria. PMID- 15126470 TI - Instability of pathogenicity islands in uropathogenic Escherichia coli 536. AB - The uropathogenic Escherichia coli strain 536 carries at least five genetic elements on its chromosome that meet all criteria characteristic of pathogenicity islands (PAIs). One main feature of these distinct DNA regions is their instability. We applied the so-called island-probing approach and individually labeled all five PAIs of E. coli 536 with the counterselectable marker sacB to evaluate the frequency of PAI-negative colonies under the influence of different environmental conditions. Furthermore, we investigated the boundaries of these PAIs. According to our experiments, PAI II536 and PAI III536 were the most unstable islands followed by PAI I536 and PAI V536, whereas PAI IV536 was stable. In addition, we found that deletion of PAI II536 and PAI III536 was induced by several environmental stimuli. Whereas excision of PAI I536, PAI II536, and PAI V536 was based on site-specific recombination between short direct repeat sequences at their boundaries, PAI III536 was deleted either by site-specific recombination or by homologous recombination between two IS100-specific sequences. In all cases, deletion is thought to lead to the formation of nonreplicative circular intermediates. Such extrachromosomal derivatives of PAI II536 and PAI III536 were detected by a specific PCR assay. Our data indicate that the genome content of uropathogenic E. coli can be modulated by deletion of PAIs. PMID- 15126471 TI - Distinctive protein signatures provide molecular markers and evidence for the monophyletic nature of the deinococcus-thermus phylum. AB - The Deinococcus-Thermus group of species is currently recognized as a distinct phylum solely on the basis of their branching in 16S rRNA trees. No unique biochemical or molecular characteristics that can distinguish this group from all other bacteria are known at present. In this work, we describe eight conserved indels (viz., inserts or deletions) in seven widely distributed proteins that are distinctive characteristics of the Deinococcus-Thermus phylum but are not found in any other group of bacteria. The identified signatures include a 7-amino-acid (aa) insert in threonyl-tRNA synthetase, 1- and 3-aa inserts in the RNA polymerase beta' subunit, a 5-aa deletion in signal recognition particle (Ffh/SR54), a 2-aa insert in major sigma factor 70 (sigma70), a 2-aa insert in seryl-tRNA synthetase (SerRS), a 1-aa insert in ribosomal protein L1, and a 2-aa insert in UvrA homologs. By using PCR primers for conserved regions, fragments of these genes were amplified from a number of Deinococcus-Thermus species, and all such fragments (except SerRS in Deinococcus proteolyticus) were found to contain the indicated signatures. The presence of these signatures in various species from all three known genera within this phylum, viz., Deinococcus, Thermus, and Meiothermus, provide evidence that they are likely distinctive characteristics of the entire phylum which were introduced in a common ancestor of this group. The signature in SerRS, which is absent in D. proteolyticus, was likely introduced after the branching of this species. Phylogenetic studies as well as the nature of the inserts in some of these proteins (viz., sigma70 and SerRS) also support a sister group relationship between the Thermus and the Meiothermus genera. The identified signatures provide strong evidence for the monophyletic nature of the Deinococcus-Thermus phylum. These molecular markers should prove very useful in the identification of new species related to this group. PMID- 15126472 TI - An extracytoplasmic-function sigma factor is involved in a pathway controlling beta-exotoxin I production in Bacillus thuringiensis subsp. thuringiensis strain 407-1. AB - Beta-exotoxin I is an insecticidal nucleotide analogue secreted by various Bacillus thuringiensis strains. In this report, we describe the characterization and transcriptional analysis of a gene cluster, designated sigW-ecfX-ecfY, that is essential for beta-exotoxin I production in B. thuringiensis subsp. thuringiensis strain 407-1. In this strain, the disruption of the sigW cluster resulted in nontoxic culture supernatants. sigW encodes a protein of 177 residues that is 97 and 94% identical to two putative RNA polymerase extracytoplasmic function-type sigma factors from Bacillus anthracis strain Ames and Bacillus cereus strain ATCC 14579, respectively. It is also 50, 30, and 26% identical to SigW from Clostridium perfringens and SigW and SigX from Bacillus subtilis, respectively. EcfX, encoded by the gene following sigW, significantly repressed the expression of sigW when both genes were overtranscribed, suggesting that it could be the anti-sigma factor of SigW. Following the loss of its curable cry plasmid, strain 407 became unable to synthesize crystal toxins, in contrast to the mutant strain 407-1(Cry-)(Pig+), which overproduced this molecule in the absence of this plasmid. Transcriptional analysis of sigW indicated that this gene was expressed during the stationary phase and only in the 407-1(Cry-)(Pig+) mutant. This suggests that in the wild type-407(Cry+) strain, beta-exotoxin I was produced from determinants located on a cry gene-bearing plasmid and that sigW is able to induce beta-exotoxin I production in B. thuringiensis in the absence of cry gene-bearing plasmids. Although the signal responsible for this activation is unknown, these results indicate that beta-exotoxin I production in B. thuringiensis can be restored or induced via an alternative pathway that requires sigW expression. PMID- 15126473 TI - Toluene 3-monooxygenase of Ralstonia pickettii PKO1 is a para-hydroxylating enzyme. AB - Oxygenases are promising biocatalysts for performing selective hydroxylations not accessible by chemical methods. Whereas toluene 4-monooxygenase (T4MO) of Pseudomonas mendocina KR1 hydroxylates monosubstituted benzenes at the para position and toluene ortho-monooxygenase (TOM) of Burkholderia cepacia G4 hydroxylates at the ortho position, toluene 3-monooxygenase (T3MO) of Ralstonia pickettii PKO1 was reported previously to hydroxylate toluene at the meta position, producing primarily m-cresol (R. H. Olsen, J. J. Kukor, and B. Kaphammer, J. Bacteriol. 176:3749-3756, 1994). Using gas chromatography, we have discovered that T3MO hydroxylates monosubstituted benzenes predominantly at the para position. TG1/pBS(Kan)T3MO cells expressing T3MO oxidized toluene at a maximal rate of 11.5 +/- 0.33 nmol/min/mg of protein with an apparent Km value of 250 microM and produced 90% p-cresol and 10% m-cresol. This product mixture was successively transformed to 4-methylcatechol. T4MO, in comparison, produces 97% p cresol and 3% m-cresol. Pseudomonas aeruginosa PAO1 harboring pRO1966 (the original T3MO-bearing plasmid) also exhibited the same product distribution as that of TG1/pBS(Kan)T3MO. TG1/pBS(Kan)T3MO produced 66% p-nitrophenol and 34% m nitrophenol from nitrobenzene and 100% p-methoxyphenol from methoxybenzene, as well as 62% 1-naphthol and 38% 2-naphthol from naphthalene; similar results were found with TG1/pBS(Kan)T4MO. Sequencing of the tbu locus from pBS(Kan)T3MO and pRO1966 revealed complete identity between the two, thus eliminating any possible cloning errors. 1H nuclear magnetic resonance analysis confirmed the structural identity of p-cresol in samples containing the product of hydroxylation of toluene by pBS(Kan)T3MO. PMID- 15126474 TI - Characterization of monospecies biofilm formation by Helicobacter pylori. AB - As all bacteria studied to date, the gastric pathogen Helicobacter pylori has an alternate lifestyle as a biofilm. H. pylori forms biofilms on glass surfaces at the air-liquid interface in stationary or shaking batch cultures. By light microscopy, we have observed attachment of individual, spiral H. pylori to glass surfaces, followed by division to form microcolonies, merging of individual microcolonies, and growth in the third dimension. Scanning electron micrographs showed H. pylori arranged in a matrix on the glass with channels for nutrient flow, typical of other bacterial biofilms. To understand the importance of biofilms to the H. pylori life cycle, we tested the effect of mucin on biofilm formation. Our results showed that 10% mucin greatly increased the number of planktonic H. pylori while not affecting biofilm bacteria, resulting in a decline in percent adherence to the glass. This suggests that in the mucus-rich stomach, H. pylori planktonic growth is favored over biofilm formation. We also investigated the effect of specific mutations in several genes, including the quorum-sensing gene, luxS, and the cagE type IV secretion gene. Both of these mutants were found to form biofilms approximately twofold more efficiently than the wild type in both assays. These results indicate the relative importance of these genes to the production of biofilms by H. pylori and the selective enhancement of planktonic growth in the presence of gastric mucin. PMID- 15126475 TI - Regulation of photosynthesis genes in Rubrivivax gelatinosus: transcription factor PpsR is involved in both negative and positive control. AB - Induction of biosynthesis of the photosystem in anoxygenic photosynthetic bacteria occurs when the oxygen concentration drops. Control of this induction takes place primarily at the transcriptional level, with photosynthesis genes expressed preferentially under anaerobic conditions. Here, we report analysis of the transcriptional control of two photosynthesis promoters, pucBA and crtI, by the PpsR factor in Rubrivivax gelatinosus. This was accomplished by analyzing the photosystem production in the wild type and in the PPSRK (ppsR::Km) mutant grown under anaerobic and semiaerobic conditions and by assessing the beta galactosidase activity of lacZ transcriptionally fused to promoters possessing the putative PpsR-binding consensus sequences. It was found that under semiaerobic conditions, inactivation of the ppsR gene resulted in overproduction of carotenoid and bacteriochlorophyll pigments, while the production of LH2 was drastically reduced. The beta-galactosidase activity showed that, in contrast to what has been found previously for Rhodobacter species, PpsR acts in R. gelatinosus as an aerobic repressor of the crtI gene while it acts as an activator for the expression of pucBA. Inspection of the putative PpsR-binding consensus sequences revealed significant differences that may explain the different levels of expression of the two genes studied. PMID- 15126477 TI - Generation of enhanced competitive root-tip-colonizing Pseudomonas bacteria through accelerated evolution. AB - A recently published procedure to enrich for efficient competitive root tip colonizers (I. Kuiper, G. V. Bloemberg, and B. J. J. Lugtenberg, Mol. Plant Microbe Interact. 14:1197-1205) after bacterization of seeds was applied to isolate efficient competitive root tip colonizers for both the dicotyledenous plant tomato and the monocotyledenous plant grass from a random Tn5luxAB mutant bank of the good root colonizer Pseudomonas fluorescens WCS365. Unexpectedly, the best-colonizing mutant, strain PCL1286, showed a strongly enhanced competitive root-tip-colonizing phenotype. Sequence analyses of the Tn5luxAB flanking regions showed that the transposon had inserted in a mutY homolog. This gene is involved in the repair of A. G mismatches caused by spontaneous oxidation of guanine. We hypothesized that, since the mutant is defective in repairing its mismatches, its cells harbor an increased number of mutations and therefore can adapt faster to the environment of the root system. To test this hypothesis, we constructed another mutY mutant and analyzed its competitive root tip colonization behavior prior to and after enrichment. As a control, a nonmutated wild type was subjected to the enrichment procedure. The results of these analyses showed (i) that the enrichment procedure did not alter the colonization ability of the wild type, (ii) that the new mutY mutant was strongly impaired in its colonization ability, but (iii) that after three enrichment cycles it colonized significantly better than its wild type. Therefore it is concluded that both the mutY mutation and the selection procedure are required to obtain an enhanced root-tip-colonizing mutant. PMID- 15126476 TI - Involvement of the C-terminal extension of the alpha polypeptide and of the PucC protein in LH2 complex biosynthesis in Rubrivivax gelatinosus. AB - The facultative phototrophic nonsulfur bacterium Rubrivivax gelatinosus exhibits several differences from other species of purple bacteria in the organization of its photosynthetic genes. In particular, the puc operon contains only the pucB and pucA genes encoding the beta and alpha polypeptides of the light-harvesting 2 (LH2) complex. Downstream of the pucBA operon is the pucC gene in the opposite transcriptional orientation. The transcription of pucBA and pucC has been studied. No pucC transcript was detected either by Northern blotting or by reverse transcription-PCR analysis. The initiation site of pucBA transcription was determined by primer extension, and Northern blot analysis revealed the presence of two transcripts of 0.8 and 0.65 kb. The half-lives of both transcripts are longer in cells grown semiaerobically than in photosynthetically grown cells, and the small transcript is the less stable. It was reported that the alpha polypeptide, encoded by the pucA gene, presents a C-terminal extension which is not essential for LH2 function in vitro. The biological role of this alanine- and proline-rich C-terminal extension in vivo has been investigated. Two mutants with C-terminal deletions of 13 and 18 residues have been constructed. Both present the two pucBA transcripts, while their phenotypes are, respectively, LH2+ and LH2-, suggesting that a minimal length of the C-terminal extension is required for LH2 biogenesis. Another important factor involved in the LH2 biogenesis is the PucC protein. To gain insight into the function of this protein in R. gelatinosus, we constructed and characterized a PucC mutant. The mutant is devoid of LH2 complex under semiaerobiosis but still produces a small amount of these antennae under photosynthetic growth conditions. This conditional phenotype suggests the involvement of another factor in LH2 biogenesis. PMID- 15126478 TI - Organization and expression of the polynucleotide phosphorylase gene (pnp) of Streptomyces: Processing of pnp transcripts in Streptomyces antibioticus. AB - We have examined the expression of pnp encoding the 3'-5'-exoribonuclease, polynucleotide phosphorylase, in Streptomyces antibioticus. We show that the rpsO pnp operon is transcribed from at least two promoters, the first producing a readthrough transcript that includes both pnp and the gene for ribosomal protein S15 (rpsO) and a second, Ppnp, located in the rpsO-pnp intergenic region. Unlike the situation in Escherichia coli, where observation of the readthrough transcript requires mutants lacking RNase III, we detect readthrough transcripts in wild-type S. antibioticus mycelia. The Ppnp transcriptional start point was mapped by primer extension and confirmed by RNA ligase-mediated reverse transcription-PCR, a technique which discriminates between 5' ends created by transcription initiation and those produced by posttranscriptional processing. Promoter probe analysis demonstrated the presence of a functional promoter in the intergenic region. The Ppnp sequence is similar to a group of promoters recognized by the extracytoplasmic function sigma factors, sigma-R and sigma-E. We note a number of other differences in rspO-pnp structure and function between S. antibioticus and E. coli. In E. coli, pnp autoregulation and cold shock adaptation are dependent upon RNase III cleavage of an rpsO-pnp intergenic hairpin. Computer modeling of the secondary structure of the S. antibioticus readthrough transcript predicts a stem-loop structure analogous to that in E. coli. However, our analysis suggests that while the readthrough transcript observed in S. antibioticus may be processed by an RNase III-like activity, transcripts originating from Ppnp are not. Furthermore, the S. antibioticus rpsO pnp intergenic region contains two open reading frames. The larger of these, orfA, may be a pseudogene. The smaller open reading frame, orfX, also observed in Streptomyces coelicolor and Streptomyces avermitilis, may be translationally coupled to pnp and the gene downstream from pnp, a putative protease. PMID- 15126479 TI - Rusty, jammed, and well-oiled hinges: Mutations affecting the interdomain region of FliG, a rotor element of the Escherichia coli flagellar motor. AB - The FliG protein is a central component of the bacterial flagellar motor. It is one of the first proteins added during assembly of the flagellar basal body, and there are 26 copies per motor. FliG interacts directly with the Mot protein complex of the stator to generate torque, and it is a crucial player in switching the direction of flagellar rotation from clockwise (CW) to counterclockwise and vice versa. A primarily helical linker joins the N-terminal assembly domain of FliG, which is firmly attached to the FliF protein of the MS ring of the basal body, to the motility domain that interacts with MotA/MotB. We report here the results of a mutagenic analysis focused on what has been called the hinge region of the linker. Residue substitutions in this region generate a diversity of phenotypes, including motors that are strongly CW biased, infrequent switchers, rapid switchers, and transiently or permanently paused. Isolation of these mutants was facilitated by a "sensitizing" mutation (E232G) outside of the hinge region that was accidentally introduced during cloning of the chromosomal fliG gene into our vector plasmid. This mutation partially interferes with flagellar assembly and accentuates the defects associated with mutations that by themselves have little phenotypic consequence. The effects of these mutations are analyzed in the context of a conformational-coupling model for motor switching and with respect to the structure of the C-terminal 70% of FliG from Thermotoga maritima. PMID- 15126480 TI - Complexity of gas vesicle biogenesis in Halobacterium sp. strain NRC-1: identification of five new proteins. AB - The genome of Halobacterium sp. strain NRC-1 contains a large gene cluster, gvpMLKJIHGFEDACNO, that is both necessary and sufficient for the production of buoyant gas-filled vesicles. Due to the resistance of gas vesicles to solubilization, only the major gas vesicle protein GvpA and a single minor protein, GvpC, were previously detected. Here, we used immunoblotting analysis to probe for the presence of gas vesicle proteins corresponding to five additional gvp gene products. Polyclonal antisera were raised in rabbits against LacZ-GvpF, GvpJ, and -GvpM fusion proteins and against synthetic 15-amino-acid peptides from GvpG and -L. Immunoblotting analysis was performed on cell lysates of wild-type Halobacterium sp. strain NRC-1, gas vesicle-deficient mutants, and purified gas vesicles, after purification of LacZ fusion antibodies on protein A and beta galactosidase affinity columns. Our results show the presence of five new gas vesicle proteins (GvpF, GvpG, GvpJ, GvpL, and GvpM), bringing the total number of proteins identified in the organelles to seven. Two of the new gas vesicle proteins are similar to GvpA (GvpJ and GvpM), and two proteins contain predicted coiled-coil domains (GvpF and GvpL). GvpL exhibited a multiplet ladder on sodium dodecyl sulfate-polyacrylamide gels indicative of oligomerization and self assembly. We discuss the possible functions of the newly discovered gas vesicle proteins in biogenesis of these unique prokaryotic flotation organelles. PMID- 15126481 TI - Arsenic resistance in Halobacterium sp. strain NRC-1 examined by using an improved gene knockout system. AB - The genome sequence of Halobacterium sp. strain NRC-1 encodes genes homologous to those responsible for conferring resistance to arsenic. These genes occur on both the large extrachromosomal replicon pNRC100 (arsADRC and arsR2M) and on the chromosome (arsB). We studied the role of these ars genes in arsenic resistance genetically by construction of gene knockouts. Deletion of the arsADRC gene cluster in a Halobacterium NRC-1 Deltaura3 strain resulted in increased sensitivity to arsenite and antimonite but not arsenate. In contrast, knockout of the chromosomal arsB gene did not show significantly increased sensitivity to arsenite or arsenate. We also found that knockout of the arsM gene produced sensitivity to arsenite, suggesting a second novel mechanism of arsenic resistance involving a putative arsenite(III)-methyltransferase. These results indicate that Halobacterium sp. strain NRC-1 contains an arsenite and antimonite extrusion system with significant differences from bacterial counterparts. Deletion analysis was facilitated by an improved method for gene knockouts/replacements in Halobacterium that relies on both selection and counterselection of ura3 using a uracil dropout medium and 5-fluoroorotic acid. The arsenite and antimonite resistance elements were shown to be regulated, with resistance to arsenic in the wild type inducible by exposure to a sublethal concentration of the metal. Northern hybridization and reverse transcription-PCR analyses showed that arsA, arsD, arsR, arsM, arsC, and arsB, but not arsR2, are inducible by arsenite and antimonite. We discuss novel aspects of arsenic resistance in this halophilic archaeon and technical improvements in our capability for gene knockouts in the genome. PMID- 15126482 TI - Evaluation of the kinetic properties of the sporulation protein SpoIIE of Bacillus subtilis by inclusion in a model membrane. AB - Starvation induces Bacillus subtilis to initiate a developmental process (sporulation) that includes asymmetric cell division to form the prespore and the mother cell. The integral membrane protein SpoIIE is essential for the prespore specific activation of the transcription factor sigmaF, and it also has a morphogenic activity required for asymmetric division. An increase in the local concentration of SpoIIE at the polar septum of B. subtilis precedes dephosphorylation of the anti-anti-sigma factor SpoIIAA in the prespore. After closure and invagination of the asymmetric septum, phosphatase activity of SpoIIE increases severalfold, but the reason for this dramatic change in activity has not been determined. The central domain of SpoIIE has been seen to self-associate (I. Lucet et al., EMBO J. 19:1467-1475, 2000), suggesting that activation of the C-terminal PP2C-like phosphatase domain might be due to conformational changes brought about by the increased local concentration of SpoIIE in the sporulating septum. Here we report the inclusion of purified SpoIIE protein into a model membrane as a method for studying the effect of local concentration in a lipid bilayer on activity. In vitro assays indicate that the membrane-bound enzyme maintains dephosphorylation rates similar to the highly active micellar state at all molar ratios of protein to lipid. Atomic force microscopy images indicate that increased local concentration does not lead to self-association. PMID- 15126483 TI - Precise excision of the large pathogenicity island, SPI7, in Salmonella enterica serovar Typhi. AB - The large pathogenicity island (SPI7) of Salmonella enterica serovar Typhi is a 133,477-bp segment of DNA flanked by two 52-bp direct repeats overlapping the pheU (phenylalanyl-tRNA) gene, contains 151 potential open reading frames, and includes the viaB operon involved in the synthesis of Vi antigen. Some clinical isolates of S. enterica serovar Typhi are missing the entire SPI7, due to its precise excision; these strains have lost the ability to produce Vi antigen, are resistant to phage Vi-II, and invade a human epithelial cell line more rapidly. Excision of SPI7 occurs spontaneously in a clinical isolate of S. enterica serovar Typhi when it is grown in the laboratory, leaves an intact copy of the pheU gene at its novel join point, and results in the same three phenotypic consequences. SPI7 is an unstable genetic element, probably an intermediate in the pathway of lateral transfer of such pathogenicity islands among enteric gram negative bacteria. PMID- 15126485 TI - Nitrite-responsive activation of the nitrate assimilation operon in Cyanobacteria plays an essential role in up-regulation of nitrate assimilation activities under nitrate-limited growth conditions. AB - NtcB of the cyanobacterium Synechococcus elongatus strain PCC 7942 is a LysR family protein that enhances expression of the nitrate assimilation operon (nirA operon) in response to the presence of nitrite, an intermediate of assimilatory nitrate reduction. Inactivation of ntcB in this cyanobacterium specifically abolishes the nitrite responsiveness of nirA operon expression, but under nitrate replete conditions (wherein negative feedback by intracellularly generated ammonium prevails over the positive effect of nitrite) activity levels of the nitrate assimilation enzymes are marginally higher in the wild-type cells than in the mutant cells, raising the issue of whether the nitrite-promoted regulation has physiological importance. On the other hand, the strains carrying ntcB expressed much higher nitrate assimilation enzyme activities under nitrate limited growth conditions than under nitrate-replete conditions whereas the ntcB deficient strains showed levels of the enzyme activities lower than those seen under the nitrate-replete conditions. Although the ntcB mutant maintained a constant cell population in a nitrate-limited chemostat when grown as a single culture, it was diluted at a rate expected for nondividing cells when mixed with the wild-type cells and subjected to nitrate limitation in the chemostat culture system. These results demonstrated that the nitrite-promoted activation of the nitrate assimilation operon is essential for up-regulation of the nitrate assimilation activities under the conditions of nitrate limitation and for competitive utilization of nitrate. PMID- 15126484 TI - Salmonella enterica serovar Typhi strains from which SPI7, a 134-kilobase island with genes for Vi exopolysaccharide and other functions, has been deleted. AB - Salmonella enterica serovar Typhi has a 134-kb island of DNA identified as salmonella pathogenicity island 7 (SPI7), inserted between pheU and 'pheU (truncated), two genes for tRNA(Phe). SPI7 has genes for Vi exopolysaccharide, for type IVB pili, for putative conjugal transfer, and for sopE bacteriophage. Pulsed-field gel electrophoresis following digestion with the endonuclease I CeuI, using DNA from a set of 120 wild-type strains of serovar Typhi assembled from several sources, identified eight strains in which the I-CeuI G fragment, which contains SPI7, had a large deletion. In addition, agglutination tests with Vi antiserum and phage typing with Vi phages show that all eight strains are Vi negative. We therefore tested these strains for deletion of SPI7 by multiplex PCR, by microarray analysis, and by sequencing of PCR amplicons. Data show that seven of the eight strains are precise deletions of SPI7: a primer pair flanking SPI7 results in a PCR amplicon containing a single pheU gene; microarrays show that all SPI7 genes are deleted. Two of the strains produce amplicons which have A derived from pheU at bp 27, while five have C derived from 'pheU at this position; thus, the position of the crossover which results in the deletion can be inferred. The deletion in the eighth strain, TYT1669, removes 175 kb with junction points in genes STY4465 and STY4664; the left junction of SPI7 and adjacent genes, as well as part of SPI7 including the viaB operon for Vi exopolysaccharide, was removed, while the right junction of SPI7 was retained. We propose that these deletions occurred during storage following isolation. PMID- 15126487 TI - ClgR, a novel regulator of clp and lon expression in Streptomyces. AB - The clp genes encoding the Clp proteolytic complex are widespread among living organisms. Five clpP genes are present in Streptomyces. Among them, the clpP1 clpP2 operon has been shown to be involved in the Streptomyces growth cycle, as a mutation blocked differentiation at the substrate mycelium step. Four Clp ATPases have been identified in Streptomyces coelicolor (ClpX and three ClpC proteins) which are potential partners of ClpP1 ClpP2. The clpC1 gene appears to be essential, since no mutant has yet been obtained. clpP1 clpP2 and clpC1 are important for Streptomyces growth, and a study of their regulation is reported here. The clpP3 clpP4 operon, which has been studied in Streptomyces lividans, is induced in a clpP1 mutant strain, and regulation of its expression is mediated via PopR, a transcriptional regulator. We report here studies of clgR, a paralogue of popR, in S. lividans. Gel mobility shift assays and DNase I footprinting indicate that ClgR binds not only to the clpP1 and clpC1 promoters, but also to the promoter of the Lon ATP-dependent protease gene and the clgR promoter itself. ClgR recognizes the motif GTTCGC-5N-GCG. In vivo, ClgR acts as an activator of clpC1 gene and clpP1 operon expression. Similarly to PopR, ClgR degradation might be ClpP dependent and could be mediated via recognition of the two carboxy-terminal alanine residues. PMID- 15126486 TI - Horizontal transfer of CS1 pilin genes of enterotoxigenic Escherichia coli. AB - CS1 is one of a limited number of serologically distinct pili found in enterotoxigenic Escherichia coli (ETEC) strains associated with disease in people. The genes for the CS1 pilus are on a large plasmid, pCoo. We show that pCoo is not self-transmissible, although our sequence determination for part of pCoo shows regions almost identical to those in the conjugative drug resistance plasmid R64. When we introduced R64 into a strain containing pCoo, we found that pCoo was transferred to a recipient strain in mating. Most of the transconjugant pCoo plasmids result from recombination with R64, leading to acquisition of functional copies of all of the R64 transfer genes. Temporary coresidence of the drug resistance plasmid R64 with pCoo leads to a permanent change in pCoo so that it is now self-transmissible. We conclude that when R64-like plasmids are transmitted to an ETEC strain containing pCoo, their recombination may allow for spread of the pCoo plasmid to other enteric bacteria. PMID- 15126488 TI - Contribution of the RpoA C-terminal domain to stimulation of the Salmonella enterica hilA promoter by HilC and HilD. AB - Expression of invasion genes in Salmonella pathogenicity island 1 (SPI-1) is mainly driven by the transcriptional activator HilA. Transcription of hilA is subject to complex control and is stimulated by the SPI-1-encoded HilC and HilD proteins. The C-terminal domain of RpoA contributes to hilA activation by HilC/D under certain inducing conditions. PMID- 15126489 TI - Genome-wide analysis of lipoprotein expression in Escherichia coli MG1655. AB - To gain insight into the cell envelope of Escherichia coli grown under aerobic and anaerobic conditions, lipoproteins were examined by using functional genomics. The mRNA expression levels of each of these genes under three growth conditions--aerobic, anaerobic, and anaerobic with nitrate--were examined by using both Affymetrix GeneChip E. coli antisense genome arrays and real-time PCR (RT-PCR). Many genes showed significant changes in expression level. The RT-PCR results were in very good agreement with the microarray data. The results of this study represent the first insights into the possible roles of unknown lipoprotein genes and broaden our understanding of the composition of the cell envelope under different environmental conditions. Additionally, these data serve as a test set for the refinement of high-throughput bioinformatic and global gene expression methods. PMID- 15126490 TI - Lipid II-mediated pore formation by the peptide antibiotic nisin: a black lipid membrane study. AB - The antibiotic peptide nisin is the first known lantibiotic that uses a docking molecule within the bacterial cytoplasmic membrane for pore formation. Through specific interaction with the cell wall precursor lipid II, nisin forms defined pores which are stable for seconds and have pore diameters of 2 to 2.5 nm. PMID- 15126491 TI - The nucleotide transporter of Caedibacter caryophilus exhibits an extended substrate spectrum compared to the analogous ATP/ADP translocase of Rickettsia prowazekii. AB - The two obligate intracellular alphaproteobacteria Rickettsia prowazekii and Caedibacter caryophilus, a human pathogen and a paramecium endosymbiont, respectively, possess transport systems to facilitate ATP uptake from the host cell cytosol. These transport proteins, which have 65% identity at the amino acid level, were heterologously expressed in Escherichia coli, and their properties were compared. The results presented here demonstrate that the caedibacter transporter had a broader substrate than the more selective rickettsial transporter. ATP analogs with modified sugar moieties, dATP and ddATP, inhibited the transport of ATP by the caedibacter transporter but not by the rickettsial transporter. Both transporters were specific for di- and trinucleotides with an adenine base in that adenosine tetraphosphate, AMP, UTP, CTP, and GTP were not competitive inhibitors. Furthermore, the antiporter nature of both transport systems was shown by the dependence of the efflux of [alpha-32P]ATP on the influx of substrate (ATP but not dATP for rickettsiae, ATP or dATP for caedibacter). PMID- 15126493 TI - Isolation and characterization of a generalized transducing phage for Pseudomonas aeruginosa strains PAO1 and PA14. AB - A temperate, type IV pilus-dependent, double-stranded DNA bacteriophage named DMS3 was isolated from a clinical strain of Pseudomonas aeruginosa. A clear plaque variant of this bacteriophage was isolated. DMS3 is capable of mediating generalized transduction within and between P. aeruginosa strains PA14 and PAO1, thus providing a useful tool for the genetic analysis of P. aeruginosa. PMID- 15126492 TI - Characterization of a nucleotide-binding domain associated with neisserial iron transport. AB - The fbpABC operon in Neisseria gonorrhoeae encodes an ATP-binding cassette transporter required for iron uptake from the host ferric binding proteins. The gene for the nucleotide-binding domain (fbpC) expressed in Escherichia coli has intrinsic ATPase activity (0.5 mmol/min/mg) uncoupled from the iron transport process. The FbpC E164D mutant is found to have a 10-fold reduction in specific activity. FbpC is covalently modified by 8-azido-[gamma32P]ATP, indicating that FbpC is a functional ATPase that likely combines with FbpB to form a ferric iron transporter. PMID- 15126494 TI - -1 frameshifting at a CGA AAG hexanucleotide site is required for transposition of insertion sequence IS1222. AB - The discovery of programmed -1 frameshifting at the hexanucleotide shift site CGA_AAG, in addition to the classical X_XXY_YYZ heptanucleotide shift sequences, prompted a search for instances among eubacterial insertion sequence elements. IS1222 has a CGA_AAG shift site. A genetic analysis revealed that frameshifting at this site is required for transposition. PMID- 15126495 TI - A phage protein confers resistance to the lactococcal abortive infection mechanism AbiP. AB - Phage bIL66M1 is sensitive to the lactococcal abortive infection mechanism AbiP. No spontaneous AbiP-resistant variant could be obtained at a frequency of <10( 10). However, AbiP-resistant variants were readily obtained during infection with both bIL66M1 and the highly homologous AbiP-resistant phage bIL170. Gain of AbiP resistance was due to the acquisition of the e6 gene from bIL170. PMID- 15126496 TI - The Escherichia coli 3-methyladenine DNA glycosylase AlkA has a remarkably versatile active site. AB - 3-Methyladenine DNA glycosylase II (AlkA) from Escherichia coli is induced in response to DNA alkylation, and it protects cells from alkylated nucleobases by catalyzing their excision. In contrast to the highly specific 3-methyladenine DNA glycosylase I (E. coli TAG) that catalyzes the excision of 3-methyl adducts of adenosine and guanosine from DNA, AlkA catalyzes the excision of a wide variety of alkylated bases including N-3 and N-7 adducts of adenosine and guanosine and O(2) adducts of thymidine and cytidine. We have investigated how AlkA can recognize a diverse set of damaged bases by characterizing its discrimination between oligonucleotide substrates in vitro. Similar rate enhancements are observed for the excision of a structurally diverse set of substituted purine bases and of the normal purines adenine and guanine. These results are consistent with a remarkably indiscriminate active site and suggest that the rate of AlkA catalyzed excision is dictated not by the catalytic recognition of a specific substrate but instead by the reactivity of the N-glycosidic bond of each substrate. Damaged bases with altered base pairing have a modest advantage, as mismatches are processed up to 400-fold faster than stable Watson-Crick base pairs. Nevertheless, AlkA does not effectively exclude undamaged DNA from its active site. The resulting deleterious excision of normal bases is expected to have a substantial cost associated with the expression of AlkA. PMID- 15126497 TI - Mutual targeting of mediator and the TFIIH kinase Kin28. AB - In Saccharomyces cerevisiae, Kin28 is a member of the cyclin-dependent kinase family. Kin28 is a subunit of the basal transcription factor holo-TFIIH and its trimeric sub-complex TFIIK. Kin28 is the primary kinase that phosphorylates the RNA polymerase II (RNA pol II) C-terminal domain (CTD) within a transcription initiation complex. Mediator, a global transcriptional co-activator, dramatically enhances the phosphorylation of the CTD of RNA pol II by holo-TFIIH in vitro. Using purified proteins we have determined that the subunits of TFIIK are sufficient for Mediator to enhance Kin28 CTD kinase activity and that Mediator enhances phosphorylation of a glutathione S-transferase-CTD fusion protein, despite the absence of multiple Mediator and/or TFIIH interactions with polymerase. Mediator does not stimulate the activity of several other CTD kinases, suggesting that the specific enhancement of TFIIH kinase activity results in Kin28 being the primary CTD kinase at initiation. In addition, we have found that Kin28 phosphorylates Mediator subunit Med4 in an assay, including purified holo-TFIIH, and either Mediator or recombinant Med4 alone. Furthermore, Kin28 appears to be, at least in part, responsible for the phosphorylation of Med4 in vivo. We have identified Thr-237 as the site of phosphorylation of Med4 by Kin28 in vitro. The mutation of Thr-237 to Ala has no effect on the growth of a yeast strain under normal conditions but confirms that Thr-237 is also the site of Med4 phosphorylation in vivo. PMID- 15126498 TI - Antimitogenesis linked to regulation of Skp2 gene expression. AB - Prostacyclin has many effects in the vasculature; one of the less well understood is the ability to block cell cycle progression through G(1) phase. We previously reported that the prostacyclin mimetic, cicaprost, selectively inhibits cyclin E cyclin-dependent kinase-2 (Cdk2), and now we show that it acts by regulating the expression of Skp2, the F-box protein that targets p27(Kip1) for ubiquitin mediated proteolysis. First, we show that cicaprost prevents the late G(1) phase down-regulation of p27(Kip1) and that the inhibitory effect of cicaprost on cyclin E-Cdk2 activity and S phase entry is eliminated by deleting p27(Kip1). Levels of the closely related Cdk2 inhibitor, p21(Cip1), are unaffected by cicaprost. Moreover, we show that cicaprost blocks the induction of Skp2 mRNA and that ectopic expression of a Skp2 cDNA overrides the effect of cicaprost on p27(Kip1) levels and S phase entry. Our data show that inhibition of F-box protein gene expression can underlie the effect of a potent antimitogen. PMID- 15126499 TI - Structural basis of membrane targeting by the Phox homology domain of cytokine independent survival kinase (CISK-PX). AB - The cytokine-independent survival kinase (CISK) in the serum and glucocorticoid regulated kinase family plays an important role in mediating cell growth and survival. N-terminal to its catalytic kinase domain, CISK contains a phox homology (PX) domain, a phosphoinositide-binding motif that directs the membrane localization of CISK and regulates CISK activity. We have determined the crystal structures of the mouse CISK-PX domain to unravel the structural basis of membrane targeting of CISK. In addition to the specific interactions conferred by the phosphoinositide-binding pocket, the structure suggests that a hydrophobic loop region and a hydrophilic beta-turn contribute to the interactions with the membrane. Furthermore, biochemical studies reveal that CISK-PX dimerizes in the presence of the linker between the PX domain and kinase domain, suggesting a multivalent mechanism in membrane localization of CISK. PMID- 15126501 TI - Identification of a trafficking motif involved in the stabilization and polarization of P2X receptors. AB - Extracellular ATP-gated channels (P2X receptors) define the third major family of ionotropic receptors, and they are expressed widely in nerve cells, muscles, and endocrine and exocrine glands. P2X subunits have two membrane-spanning domains, and a receptor is thought to be formed by oligomerization of three subunits. We have identified a conserved motif in the cytoplasmic C termini of P2X subunits that is necessary for their surface expression; mutations in this motif result in a marked reduction of the receptors at the plasma membrane because of a rapid internalization. Transfer of the motif to a reporter protein (CD(4)) enhances the surface expression of the chimera, indicating that this motif is likely involved in the stabilization of P2X receptor at the cell surface. In neurons, mutated P2X(2) subunits showed reduced membrane expression and an altered axodendritic distribution. This motif is also present in intracellular regions of other membrane proteins, such as in the third intracellular loop of some G protein coupled receptors, suggesting that it might be involve in their cellular stabilization and polarization. PMID- 15126500 TI - YIH1 is an actin-binding protein that inhibits protein kinase GCN2 and impairs general amino acid control when overexpressed. AB - The general amino acid control (GAAC) enables yeast cells to overcome amino acid deprivation by activation of the alpha subunit of translation initiation factor 2 (eIF2alpha) kinase GCN2 and consequent induction of GCN4, a transcriptional activator of amino acid biosynthetic genes. Binding of GCN2 to GCN1 is required for stimulation of GCN2 kinase activity by uncharged tRNA in starved cells. Here we show that YIH1, when overexpressed, dampens the GAAC response (Gcn- phenotype) by suppressing eIF2alpha phosphorylation by GCN2. The overexpressed YIH1 binds GCN1 and reduces GCN1-GCN2 complex formation, and, consistent with this, the Gcn- phenotype produced by YIH1 overexpression is suppressed by GCN2 overexpression. YIH1 interacts with the same GCN1 fragment that binds GCN2, and this YIH1-GCN1 interaction requires Arg-2259 in GCN1 in vitro and in full-length GCN1 in vivo, as found for GCN2-GCN1 interaction. However, deletion of YIH1 does not increase eIF2alpha phosphorylation or derepress the GAAC, suggesting that YIH1 at native levels is not a general inhibitor of GCN2 activity. We discovered that YIH1 normally resides in a complex with monomeric actin, rather than GCN1, and that a genetic reduction in actin levels decreases the GAAC response. This Gcn- phenotype was partially suppressed by deletion of YIH1, consistent with YIH1 mediated inhibition of GCN2 in actin-deficient cells. We suggest that YIH1 resides in a YIH1-actin complex and may be released for inhibition of GCN2 and stimulation of protein synthesis under specialized conditions or in a restricted cellular compartment in which YIH1 is displaced from monomeric actin. PMID- 15126502 TI - Anti-chemorepulsive effects of vascular endothelial growth factor and placental growth factor-2 in dorsal root ganglion neurons are mediated via neuropilin-1 and cyclooxygenase-derived prostanoid production. AB - Vascular endothelial growth factor (VEGF) displays neurotrophic and neuroprotective activities, but the mechanisms underlying these effects have not been defined. Neuropilin-1 (NP-1) is a receptor for VEGF165 and placental growth factor-2 (PlGF-2), but the role of NP-1 in VEGF-dependent neurotrophic actions is unclear. Dorsal root ganglion (DRG) neurons expressed high levels of NP-1 mRNA and protein, much lower levels of KDR, and no detectable Flt-1. VEGF165 and PlGF 2 promoted DRG growth cone formation with an effect similar to that of nerve growth factor, whereas the Flt-1-specific ligand, PlGF-1, and the KDR/Flt-4 ligand, VEGF-D, had no effect. The chemorepellent NP-1 ligand, semaphorin 3A, antagonized the response to VEGF and PlGF-2. The specific KDR inhibitor, SU5614, did not affect the anti-chemorepellent effects of VEGF and PlGF-2, whereas a novel, specific antagonist of VEGF binding to NP-1, called EG3287, prevented inhibition of growth cone collapse. VEGF stimulated prostacyclin and prostaglandin E2 production in DRG cultures that was blocked by inhibitors of cyclooxygenases; the anti-chemorepellent activities of VEGF and PlGF-2 were abrogated by cyclooxygenase inhibitors, and a variety of prostacyclin analogues and prostaglandins strikingly inhibited growth cone collapse. These findings support a specific role for NP-1 in mediating neurotrophic actions of VEGF family members and also identify a novel role for prostanoids in the inhibition of neuronal chemorepulsion. PMID- 15126503 TI - Interaction between glucose-regulated destruction domain of DNA topoisomerase IIalpha and MPN domain of Jab1/CSN5. AB - DNA topoisomerase (topo) IIalpha, an essential enzyme for cell proliferation, is targeted to a proteasome-dependent degradation pathway when human tumor cells are glucose-starved. Here we show that the topo IIalpha destabilization depends on the newly identified domain, GRDD (glucose-regulated destruction domain), which was mapped to the N-terminal 70-170 amino acid sequence. Indeed, the deletion of GRDD conferred a stable feature on topo IIalpha, whereas the fusion of GRDD rendered green fluorescent protein unstable under glucose starvation conditions. Nuclear localization was a prerequisite for GRDD function, because the inhibition of nuclear translocation resulted in the suppression of GRDD-mediated topo IIalpha degradation. Further, GRDD was identified as an interactive domain for Jab1/CSN5, which promoted the degradation of topo IIalpha in a manner dependent on the MPN (Mpr1p/Prd1p N terminus) domain. Depleting Jab1/CSN5 by antisense oligonucleotide and treating cells with the CSN-associated kinase inhibitor, curcumin, inhibited topo IIalpha degradation induced by glucose starvation. These findings demonstrate that GRDD can act as a stress-activated degron for regulating topo IIalpha stability, possibly through interaction with the MPN domain of Jab1/CSN5. PMID- 15126504 TI - Down-regulation of WW domain-containing oxidoreductase induces Tau phosphorylation in vitro. A potential role in Alzheimer's disease. AB - Numerous enzymes hyperphosphorylate Tau in vivo, leading to the formation of neurofibrillary tangles (NFTs) in the neurons of Alzheimer's disease (AD). Compared with age-matched normal controls, we demonstrated here that the protein levels of WW domain-containing oxidoreductase WOX1 (also known as WWOX or FOR), its Tyr33-phosphorylated form, and WOX2 were significantly down-regulated in the neurons of AD hippocampi. Remarkably knock-down of WOX1 expression by small interfering RNA in neuroblastoma SK-N-SH cells spontaneously induced Tau phosphorylation at Thr212/Thr231 and Ser515/Ser516, enhanced phosphorylation of glycogen synthase kinase 3beta (GSK-3beta) and ERK, and enhanced NFT formation. Also an increased binding of phospho-GSK-3beta with phospho-Tau was observed in these WOX1 knock-down cells. In comparison, increased phosphorylation of Tau, GSK 3beta, and ERK, as well as NFT formation, was observed in the AD hippocampi. Activation of JNK1 by anisomycin further increased Tau phosphorylation, and SP600125 (a JNK inhibitor) and PD-98059 (an MEK1/2 inhibitor) blocked Tau phosphorylation and NFT formation in these WOX1 knock-down cells. Ectopic or endogenous WOX1 colocalized with Tau, JNK1, and GSK-3beta in neurons and cultured cells. 17Beta-estradiol, a neuronal protective hormone, increased the binding of WOX1 and GSK-3beta with Tau. Mapping analysis showed that WOX1 bound Tau via its COOH-terminal short-chain alcohol dehydrogenase/reductase domain. Together WOX1 binds Tau via its short-chain alcohol dehydrogenase/reductase domain and is likely to play a critical role in regulating Tau hyperphosphorylation and NFT formation in vivo. PMID- 15126505 TI - Evidence for differing roles for each lobe of the calmodulin-like domain in a calcium-dependent protein kinase. AB - Calcium-dependent protein kinases (CDPKs) are structurally unique Ser/Thr kinases found in plants and certain protozoa. They are distinguished by a calmodulin-like regulatory apparatus (calmodulin-like domain (CaM-LD)) that is joined via a junction (J) region to the C-terminal end of the kinase catalytic domain. Like CaM, the CaM-LD is composed of two globular EF structural domains (N-lobe, C lobe), each containing a pair of Ca(2+) binding sites. Spectroscopic analysis shows that the CaM-LD is comprised of helical elements, but the isolated CaM-LD does not form a conformationally homogeneous tertiary structure in the absence of Ca(2+). The addition of substoichiometric amounts of Ca(2+) is sufficient to stabilize the C-terminal lobe in a construct containing J and CaM-LD (JC) but not in the CaM-LD alone. Moreover, as J is titrated into Ca(2+)-saturated CaM-LD, interactions are stronger with the C-lobe than the N-lobe of the CaM-LD. Measurements of Ca(2+) affinity for JC reveal two cooperatively interacting high affinity binding sites (K(d)(,mean) = 5.6 nm at 20 mm KCl) in the C-lobe and two weaker sites in the N-lobe (K(d,mean) = 110 nm at 20 mm KCl). The corresponding Ca(2+) binding constants in the isolated CaM-LD are lower by more than 2 orders of magnitude, which indicates that the J region has an essential role in stabilizing the structure of the CDPK regulatory apparatus. The large differential affinity between the two domains together with previous studies on a plasmodium CDPK (Zhao, Y., Pokutta, S., Maurer, P., Lindt, M., Franklin, R. M., and Kappes, B. (1994) Biochemistry 33, 3714-3721) suggests a model whereby even at normally low cytosolic levels of Ca(2+), the C-lobe interacts with the junction, but the kinase remains in an autoinhibited state. Activation then occurs when Ca(2+) levels rise to fill the two weaker affinity binding sites in the N-lobe, thereby triggering a conformational change that leads to release of the autoinhibitory region. PMID- 15126506 TI - FOXO4 is acetylated upon peroxide stress and deacetylated by the longevity protein hSir2(SIRT1). AB - FOXO transcription factors have important roles in metabolism, cellular proliferation, stress tolerance, and aging. FOXOs are negatively regulated by protein kinase B/c-Akt-mediated phosphorylation. Here we show that FOXO factors are also subject to regulation by reversible acetylation. We provide evidence that the acetyltransferase CREB-binding protein (CBP) binds FOXO resulting in acetylation of FOXO. This acetylation inhibits FOXO transcriptional activity. Binding of CBP and acetylation are induced after treatment of cells with peroxide stress. Deacetylation of FOXOs involves binding of the NAD-dependent deacetylase hSir2(SIRT1). Accordingly, hSir2(SIRT1)-mediated deacetylation precludes FOXO inhibition through acetylation and thereby prolongs FOXO-dependent transcription of stress-regulating genes. These data demonstrate that acetylation functions in a second pathway of negative control for FOXO factors and provides a novel mechanism whereby hSir2(SIRT1) can promote cellular survival and increase lifespan. PMID- 15126508 TI - The low density lipoprotein receptor-related protein 1B retains beta-amyloid precursor protein at the cell surface and reduces amyloid-beta peptide production. AB - The low density lipoprotein (LDL) receptor-related protein 1B (LRP1B) is a newly identified member of the LDL receptor family that shares high homology with the LDL receptor-related protein (LRP). LRP1B was originally described as a putative tumor suppressor in lung cancer cells; however, its expression profile in several regions of adult human brain suggests it may have additional functions in the central nervous system. Since LRP1B has overlapping ligand binding properties with LRP, we investigated whether LRP1B, like LRP, could interact with the beta amyloid precursor protein (APP) and modulate its processing to amyloid-beta peptides (Abetas). Using an LRP1B minireceptor (mLRP1B4) generated to study the trafficking of LRP1B, we found that mLRP1B4 and APP form an immunoprecipitable complex. Furthermore mLRP1B4 bound and facilitated the degradation of a soluble isoform of APP containing a Kunitz proteinase inhibitor domain but not soluble APP lacking a Kunitz proteinase inhibitor domain. A functional consequence of mLRP1B4 expression was a significant accumulation of APP at the cell surface, which is likely related to the slow endocytosis rate of LRP1B. More importantly, mLRP1B4-expressing cells that accumulated cell surface APP produced less Abeta and secreted more soluble APP. These findings reveal that LRP1B is a novel binding partner of APP that functions to decrease APP processing to Abeta. Consequently LRP1B expression could function to protect against the pathogenesis of Alzheimer's disease. PMID- 15126507 TI - The heptahelical domain of GABA(B2) is activated directly by CGP7930, a positive allosteric modulator of the GABA(B) receptor. AB - The gamma-aminobutyric acid, type B (GABA(B)) receptor is well recognized as being composed of two subunits, GABA(B1) and GABA(B2). Both subunits share structural homology with other class-III G-protein-coupled receptors. They are composed of two main domains: a heptahelical domain (HD) typical of all G-protein coupled receptors and a large extracellular domain (ECD). Although GABA(B1) binds GABA, GABA(B2) is required for GABA(B1) to reach the cell surface. However, it is still not demonstrated whether the association of these two subunits is always required for function in the brain. Indeed, GABA(B2) plays a major role in the coupling of the heteromer to G-proteins, such that it is possible that GABA(B2) can transmit a signal in the absence of GABA(B1). Today only ligands interacting with GABA(B1) ECD have been identified. Thus, the compounds acting exclusively on the GABA(B2) subunit will be helpful in analyzing the specific role of this subunit in the brain. Here, we explored the mechanism of action of CGP7930, a compound described as a positive allosteric regulator of the GABA(B) receptor. We showed that it activates the wild type GABA(B) receptor but with a low efficacy. The GABA(B2) HD is necessary for this effect, although one cannot exclude that CGP7930 could also bind to GABA(B1). Of interest, CGP7930 could activate GABA(B2) expressed alone and is the first described agonist of GABA(B2). Finally, we show that CGP7930 retains its agonist activity on a GABA(B2) subunit deleted of its ECD. This demonstrates that the HD of GABA(B2) behaves similar to a rhodopsin like receptor, because it can reach the cell surface alone, can couple to G protein, and be activated by agonists. These data open new strategies for studying the mechanism of activation of GABA(B) receptor and examine any possible role of homomeric GABA(B2) receptors. PMID- 15126509 TI - MAZ elements alter transcription elongation and silencing of the fibroblast growth factor receptor 2 exon IIIb. AB - The fibroblast growth factor receptor 2 (FGFR2) gene exons IIIb and IIIc are alternatively spliced in a mutually exclusive and cell type-specific manner. FGFR2 exon choice depends on both activation and silencing. Exon IIIb silencing requires cis-acting elements upstream and downstream of the exon. To examine the influence of transcription on exon IIIb silencing, the putative RNA polymerase II (RNAPII)-pausing MAZ4 element was inserted at different positions within the FGFR2 minigene construct. MAZ4 insertions 5' to the upstream silencing elements or between exon IIIb and downstream silencing elements result in decreased silencing. An insertion 3' of the downstream silencing elements, however, has no effect on splicing. An RT-PCR elongation assay shows that the MAZ4 site in these constructs is likely to be a RNAPII pause site. Insertion of another RNAPII pause site into the minigene has a similar effect on exon IIIb silencing. Transfection of in vitro transcribed RNA demonstrates that the cell type specificity of FGFR2 alternative splicing requires co-transcriptional splicing. Additionally, changing the promoter alters both FGFR2 minigene splicing and the MAZ4 effect. We propose that RNAPII pauses at the MAZ4 elements resulting in a change in the transcription elongation complex that influences alternative splicing decisions downstream. PMID- 15126510 TI - Care of common endocrine problems in Cuba. PMID- 15126511 TI - Novel target genes for catecholamines in skeletal muscle. PMID- 15126513 TI - Hepatic lipase genotype, diabetes risk, and implications for preventative medicine. PMID- 15126512 TI - In vivo epinephrine-mediated regulation of gene expression in human skeletal muscle. AB - The stress hormone epinephrine produces major physiological effects on skeletal muscle. Here we determined skeletal muscle mRNA expression profiles before and during a 6-h epinephrine infusion performed in nine young men. Stringent statistical analysis of data obtained using 43000 cDNA element microarrays showed that 1206 and 474 genes were up- and down-regulated, respectively. Microarray data were validated using reverse transcription quantitative PCR. Gene classification was performed through data mining of Gene Ontology annotations, cluster analysis of regulated genes among 14 human tissues, and correlation analysis of mRNA and clinical parameter variations. Evidence of an autoregulatory control was provided by the regulation of key genes of the cAMP-dependent transcription pathway. Genes with known functional cAMP response elements were regulated by the hormone. The impact on metabolism was illustrated by coordinated regulations of genes involved in carbohydrate and protein metabolisms. Epinephrine had a profound effect on genes involved in immunity and inflammatory response, a previously unappreciated aspect of catecholamine action. Information on 526 mRNAs corresponded to genes of unknown function. These data define the molecular signatures of epinephrine action in human skeletal muscle. They may contribute to the understanding of skeletal muscle alterations observed in pathological conditions characterized by sympathetic nervous system overdrive. PMID- 15126514 TI - The G-250A promoter polymorphism of the hepatic lipase gene predicts the conversion from impaired glucose tolerance to type 2 diabetes mellitus: the Finnish Diabetes Prevention Study. AB - In population-based studies, dyslipidemia related to insulin resistance (high triglyceride level and low high-density lipoprotein cholesterol level) is a risk factor for type 2 diabetes. Therefore, variants in genes regulating lipid and lipoprotein metabolism are potential candidate genes for diabetes. We investigated whether the G-250A polymorphism of the hepatic lipase gene (LIPC) predicts the conversion from impaired glucose tolerance (IGT) to type 2 diabetes in the Finnish Diabetes Prevention Study. This study randomized subjects to either the intervention group (lifestyle modification aimed at weight loss, such as changes in diet and increased physical exercise) or the control group. Genotyping at position -250 of the LIPC gene was performed with PCR amplification, DraI enzyme digestion, and gel electrophoresis in 490 subjects with IGT whose DNA was available. In the entire study population, the conversion rate to type 2 diabetes was 17.8% among subjects with the G-250G genotype and 10.7% among subjects with the -250A allele (P = 0.032). In univariate analysis, the odds ratio for the G-250G genotype to predict the conversion from IGT to type 2 diabetes was 1.80 (95% confidence interval, 1.05-3.10; P = 0.034). In multivariate logistic regression analysis, the G-250G genotype predicted the conversion to diabetes independently of the study group (control or intervention), gender, weight, waist circumference at baseline, and change in weight and waist circumference. In the intervention group, 13.0% of subjects with the G-250G genotype and 1.0% of the subjects with the -250A allele converted to diabetes (P = 0.001). We conclude that the G-250G genotype of the LIPC gene is a risk factor for type 2 diabetes. Therefore, genes regulating lipid and lipoprotein metabolism may be potential candidate genes for type 2 diabetes. PMID- 15126515 TI - In vitro expression studies of a novel mutation delta299 in a patient affected with apparent mineralocorticoid excess. AB - Apparent mineralocorticoid excess syndrome (AME) is an autosomal recessive disorder that results in low renin hypertension and other characteristic clinical features. Typical patients present with severe hypertension, hypokalemia, and undetectable aldosterone. Most patients also have low birth weight, failure to thrive, and nephrocalcinosis. The 11betahydroxysteroid dehydrogenase type 2 (11betaHSD2) defect is documented by demonstrating a failure to convert cortisol to cortisone. Here, we report a patient with typical phenotypic features of AME who does not carry any of the previously described mutations in the HSD11B2 gene. This female patient from a consanguineous Pakistani family presented at age 9 yr. She had a low birth weight compared with her siblings and presented with hypertension (225/120 mm Hg), low plasma renin activity, hypokalemic metabolic alkalosis, suppressed aldosterone, and bilateral nephrocalcinosis. Echocardiogram did not reveal left ventricular hypertrophy, and baseline ophthalmological evaluation did not demonstrate hypertensive retinopathy. However, at age 12 yr, she developed mild to moderate hypertensive retinopathy. Biochemical analysis showed an elevated urinary cortisol to cortisone metabolites ratio (tetrahydrocortisol and 5alpha-tetrahydrocortisol/tetrahydrocortisone) of 28 (normal, 0.66-2.44). She had a cortisol secretion rate of 0.43 mg/d (normal, 5-25 mg/d). Sequence analysis of the HSD11B2 gene revealed a novel homozygous delta299 mutation in exon 5. In vitro expression in Chinese hamster ovary cells revealed that this mutation resulted in no activity. PMID- 15126516 TI - A homozygous null mutation delineates the role of the melanocortin-4 receptor in humans. AB - As a mediator of the effects of leptin, the melanocortin-4 receptor (MC4R) is an essential component of the central regulation of long-term energy homeostasis. Heterozygous mutations in this receptor are the most frequent genetic cause of severe obesity in children. The very rare described carriers of homozygous MC4R mutations for whom clinical data were available had a residual receptor activity thus not allowing for the description of the full extent of the role of MC4R in humans. Here, we present the clinical and biological features of a patient with complete absence of MC4R activity and compare the clinical and endocrine characteristics of this patient with those previously observed in leptin receptor deficient patients. Our data suggest that in humans, the MC4R mediates most of the anorectic effects of leptin in early childhood. In contrast, MC4R does not mediate the effect of leptin on linear growth and other endocrine axes. In addition, complete MC4R deficiency is not a cause of relative hyperinsulinemia as recently observed in children with heterozygous MC4R mutations. PMID- 15126517 TI - Obstructive sleep apnea presenting as pseudopheochromocytoma: a case report. AB - Sudden arousal from sleep causes a transient surge in sympathetic nervous activity. Repeated arousals, as occur in obstructive sleep apnea (OSA), are well documented to cause a more prolonged sympathetic overactivity and consequent elevations in 24-h urinary catecholamine levels. We describe here a series of five patients, each presenting with a clinical and biochemical picture indistinguishable from that of pheochromocytoma. Thorough investigations have failed to find catecholamine-secreting tumor in any of these subjects, but all have been diagnosed with OSA. Primary treatment of OSA with nasal continuous positive airways pressure has led to normalization of systemic blood pressure and urinary catecholamines. Pseudopheochromocytoma is therefore a rare, but treatable, presentation of obstructive sleep apnea. PMID- 15126518 TI - A comparison of a novel testosterone bioadhesive buccal system, striant, with a testosterone adhesive patch in hypogonadal males. AB - A novel delivery system has been developed for testosterone replacement. This formulation, COL-1621 (Striant), a testosterone-containing buccal mucoadhesive system, has been shown in preliminary studies to replace testosterone at physiological levels when used twice daily. Therefore, the current study compared the steady-state pharmacokinetics and tolerability of the buccal system with a testosterone-containing skin patch (Andropatch or Androderm) in an international multicenter study of a group of hypogonadal men. Sixty-six patients were randomized into two groups; one applied the buccal system twice daily, whereas the other applied the transdermal patch daily, in each case for 7 d. Serum total testosterone and dihydrotestosterone concentrations were measured at d 1, 3 or 4, and 6, and serially over the last 24 h of the study. Pharmacokinetic parameters for each formulation were calculated, and the two groups were compared. The tolerability of both formulations was also evaluated. Thirty-three patients were treated with the buccal preparation, and 34 were treated with the transdermal patch. The average serum testosterone concentration over 24 h showed a mean of 18.74 nmol/liter (SD =; 5.90) in the buccal system group and 12.15 nmol/liter (SD =; 5.55) in the transdermal patch group (P < 0.01). Of the patients treated with the buccal system, 97% had average steady-state testosterone concentrations within the physiological range (10.41-36.44 nmol/liter), whereas only 56% of the transdermal patch patients achieved physiological total testosterone concentrations (P < 0.001 between groups). Testosterone concentrations were within the physiological range in the buccal system group for a significantly greater portion of the 24-h treatment period than in the transdermal patch group (mean, 84.9% vs. 54.9%; P < 0.001). Testosterone/dihydrotestosterone ratios were physiological and similar in both groups. Few patients experienced major adverse effects from either treatment. No significant local tolerability problems were noted with the buccal system, other than a single patient withdrawal. We conclude that this buccal system is superior to the transdermal patch in achieving testosterone concentrations within the normal range. It may, therefore, be a valuable addition to the range of choices for testosterone replacement therapy. PMID- 15126519 TI - The association of the K121Q polymorphism of the plasma cell glycoprotein-1 gene with type 2 diabetes and hypertension depends on size at birth. AB - Birth weight and length serve as indicators of the intrauterine environment, and a small body size at birth is a predictor of type 2 diabetes and hypertension. Insulin is one of the growth factors regulating fetal growth. The plasma cell glycoprotein 1 (PC-1) gene impairs insulin signaling at the insulin receptor level. Therefore, we investigated whether the K121Q polymorphism of the PC-1 gene association with insulin sensitivity, insulin levels, and the prevalence of diabetes and hypertension in adult life depends on size at birth in 489 subjects born in Helsinki during 1924-1933. We found that the effect of the PC-1 gene polymorphism on insulin levels and insulin sensitivity, measured as the homeostasis model assessment for insulin resistance, depended on birth length because fasting insulin levels and insulin resistance were highest in subjects carrying the 121Q allele who were small at birth (P for interaction = 0.04 and 0.05). Additionally, in those whose birth length was up to 49 cm, the K121Q polymorphism of the PC-1 gene was associated with a 2-fold higher incidence of type 2 diabetes. Moreover, subjects who were short at birth and who had the 121Q allele had the highest incidence (31.6%) of type 2 diabetes together with hypertension. We conclude that the interaction between the K121Q polymorphism of the PC-1 gene and birth length affects insulin sensitivity and increases susceptibility to type 2 diabetes and hypertension in adulthood. PMID- 15126520 TI - Growth hormone (GH) replacement therapy in adult-onset gh deficiency: effects on body composition in men and women in a double-blind, randomized, placebo controlled trial. AB - Adult GH deficiency (AGHD) is characterized by an altered body composition, an atherogenic lipid profile, decreased exercise capacity, and diminished quality of life. We performed a randomized, double-blind, placebo-controlled, multicenter study in 166 subjects with AGHD to assess the effects of GH on these outcomes. GH was initiated at 0.0125 mg/kg.d, increased to 0.025 mg/kg.d as tolerated, or decreased to 0.00625 mg/kg.d for 12 months. Primary measures of efficacy included body composition, strength and endurance, and quality of life. Additional parameters included serum IGF-I concentrations, serum lipids, and bone mineral density. After 12 months, 79% of subjects remained on GH 0.0125 mg/kg.d, whereas 21% received 0.00625 mg/kg.d. GH-treated men and women demonstrated significant decreases in total body and trunk fat and increases in lean body mass over baseline. In GH-treated men, mean IGF-I SD scores exceeded age-adjusted normal ranges, whereas similar doses produced a smaller response in women. GH treatment was associated with significant improvements in total cholesterol and low-density lipoprotein (P < 0.05 for all). No significant treatment effects were observed in strength and endurance, quality of life, or bone mineral density. GH treatment was generally well tolerated. Subjects with AGHD should receive individualized GH therapy to maintain IGF-I between the mean value and +2 SD and improve body composition and cardiovascular risk factors. PMID- 15126521 TI - Plasma metanephrine levels are decreased in type 1 diabetic patients with a severely impaired epinephrine response to hypoglycemia, indicating reduced adrenomedullary stores of epinephrine. AB - A defective epinephrine response to hypoglycemia is a common disorder in type 1 diabetes. We assessed the role of the adrenomedullary capacity to secrete epinephrine in this disorder by measuring plasma metanephrine levels in affected type 1 diabetic patients compared with those in matched nondiabetic controls. Metanephrine is formed from epinephrine that leaks from adrenomedullary storage vesicles by catechol-O-methyl transferase (COMT) and is continuously released into the circulation. Thus, plasma metanephrine levels reflect adrenomedullary epinephrine content and, provided there is normal COMT activity, the adrenomedullary capacity to secrete epinephrine. Diabetic patients had approximately 25% lower plasma metanephrine levels than controls (0.18 +/- 0.09 vs. 0.24 +/- 0.02 nmol/liter; P = 0.012), whereas plasma epinephrine, norepinephrine, and normetanephrine levels were comparable between patients and controls. In response to hypoglycemia, the increments in plasma epinephrine and plasma metanephrine levels were both significantly lower in diabetic patients than in controls (P < 0.001), but the increase in plasma metanephrine as a percentage of the increase in plasma epinephrine was identical, indicating similar COMT activity. We conclude that type 1 diabetic patients with an impaired epinephrine response to hypoglycemia have lower plasma metanephrine levels than matched controls, reflecting decreased adrenomedullary stores of epinephrine and indicating reduced adrenomedullary capacity to secrete epinephrine. PMID- 15126522 TI - Induction of multiple follicular development by a single dose of long-acting recombinant follicle-Stimulating hormone (FSH-CTP, corifollitropin alfa) for controlled ovarian stimulation before in vitro fertilization. AB - In a first feasibility study, the efficacy and safety of a single dose of recombinant long-acting FSH (FSH-CTP) were investigated in in vitro fertilization (IVF) patients undergoing controlled ovarian stimulation with a flexible GnRH antagonist protocol. Eligible subjects were randomized to receive a single dose of 120 micro g (n = 25), 180 microg (n = 24), or 240 microg (n = 25) corifollitropin alfa (FSH-CTP) or to start daily fixed doses of 150 IU recombinant FSH (rFSH) (n = 24, reference). Subjects who received a single dose of FSH-CTP continued 1 wk after injection (treatment d 8) with fixed daily doses of 150 IU rFSH (Puregon/Follistim) until the day of triggering final oocyte maturation. The terminal half-life of FSH-CTP was, on average, 65 h and dose independent. Cycle cancellation before human chorionic gonadotropin (hCG) administration occurred in only three subjects treated with FSH-CTP. The median duration of stimulation was 10.0 d in each FSH-CTP group and 9.0 d in the daily rFSH group. The total number of follicles at least 11 mm at stimulation d 8 and at the day of hCG administration tended to increase with dose of FSH-CTP, although a significant dose-response relationship was revealed only for the number of follicles at least 15 mm on the day of hCG (P = 0.03). Serum estradiol levels and inhibin-B levels were not significantly different between the four groups on d 8 and on the day of hCG. In total, 12 subjects (17.6%) in the FSH-CTP groups and two subjects (8.3%) in the rFSH group experienced a premature LH rise (defined as LH >or= 10 IU/liter) before the start of the GnRH antagonist (P value not significant between groups). This relatively high incidence of women demonstrating an early LH rise in the FSH-CTP groups may be related to the higher initial rises of serum estradiol and the use of a flexible GnRH antagonist protocol. The mean number of oocytes recovered per started cycle was higher in FSH-CTP-treated subjects compared with rFSH-treated subjects (significant at P = 0.03 for the 240- microg FSH-CTP group), but no difference could be noted between the number of good quality embryos (range of means, 3.8-4.8 per attempt), and equal numbers of embryos were available for embryo transfer. In summary, FSH-CTP appeared to be a potent inducer of multiple follicular growth; additional research will be needed to select the optimal FSH-CTP dose and treatment time interval. PMID- 15126523 TI - Genetic and environmental causes of individual differences in thyroid size: a study of healthy Danish twins. AB - Factors such as iodine intake, serum TSH concentration, gender, age, body mass index, parity, and cigarette smoking are thought to influence thyroid size. The purpose of our study was to determine the relative roles of these environmental and physiological factors compared with genetic factors in euthyroid subjects with a clinically normal thyroid gland. A representative sample of self-reported healthy twin pairs was identified through the Danish Twin Registry. A total of 520 individuals divided into 104 monozygotic (MZ), 107 dizygotic same sex (DZ), and 49 opposite sex twin pairs were investigated. After adjustment for age, gender, and other covariates, intraclass correlations were calculated. To elucidate the relative importance of genetic and environmental factors to the variation of ultrasonically determined thyroid volume, quantitative genetic modeling was used. Regression analysis suggested that serum TSH, serum free T(4), gender, age, smoking, and body mass index each played a small, but significant, role for variation in thyroid volume. The intraclass correlations for thyroid volume were consistently higher for MZ than for DZ twin pairs (r(MZ) = 0.71; r(DZ) = 0.18; P < 0.001). Using quantitative genetic modeling, it was calculated that genetic factors (with 95% confidence intervals) accounted for 71% (61-78%) of the individual differences in thyroid volume. Genetic influences are important in the regulation of normal thyroid size. This fits the observation that goiter may be seen also in the absence of evident environmental goitrogens such as iodine deficiency and that not all individuals develop goiter even in iodine deficient areas. PMID- 15126524 TI - Inhibition of dipeptidyl peptidase-4 reduces glycemia, sustains insulin levels, and reduces glucagon levels in type 2 diabetes. AB - The stimulation of insulin vs. inhibition of glucagon secretion in relation to the antidiabetic action of glucagon-like peptide-1 (GLP-1) is not established. Here, the influence of a 4-wk increase in circulating GLP-1 by inhibition of dipeptidyl peptidase-4 (DPP-4) on 24-h glucose and insulin and glucagon responses to breakfast was studied in subjects with dietary controlled diabetes [age: 65 +/ 8 yr (SD), body mass index: 27.3 +/- 3.3 kg/m(2), fasting plasma glucose: 9.0 +/ 1.3 mmol/liter]. Compared with placebo (n = 19), a specific DPP-4 inhibitor [(1 [[(3-hydroxy-1-adamantyl) amino] acetyl]-2-cyano-(S)-pyrrolidine) (LAF237); 100 mg daily, n = 18] reduced fasting glucose by 0.70 mmol/liter (P = 0.037), 4-h prandial glucose excursion by 1.45 mmol/liter (P < 0.001), and mean 24-h glucose by 0.93 mmol/liter (P < 0.001). Baseline and postprandial active GLP-1 were increased by LAF237. The glucagon response to breakfast was reduced by LAF237 (glucagon levels at 60 min were 88 +/- 8 pg/ml before treatment vs. 77 +/- 5 pg/ml after; P = 0.001). In contrast, the overall insulin levels were not altered. The 4-wk reduction in glucagon correlated with the reduction in 2-h glucose (r = 0.61; P = 0.008). No such association was observed for insulin. Thus, improved metabolic control by DPP-4 inhibition in type 2 diabetes is seen in association with reduced glucagon levels and, despite the lower glycemia, unaltered insulin levels. PMID- 15126525 TI - Long-term testosterone gel (AndroGel) treatment maintains beneficial effects on sexual function and mood, lean and fat mass, and bone mineral density in hypogonadal men. AB - Transdermal testosterone (T) delivery represents an effective alternative to injectable androgens. We studied 163 hypogonadal men who applied 5, 7.5, or 10 g AndroGel (T gel) 1% CIII per day for up to 42 months. Efficacy data were presented in 123 subjects considered evaluable. Continuous AndroGel treatment normalized mean serum T and free T levels. Mean serum 5alpha-dihydrotestosterone concentrations and 5alpha-dihydrotestosterone/T ratio slightly increased, mean serum estradiol/T ratio doubled, and mean serum FSH and LH levels were suppressed by T replacement. Sexual function and mood parameters improved rapidly and were maintained throughout T treatment. Lean body mass increased (P = 0.0001) and fat mass decreased (P = 0.0001), and these changes were maintained with treatment but were not accompanied by significant increases in muscle strength. Increases in serum bone markers suggestive of increased bone formation were followed by gradual and progressive increases in bone mineral density more in the spine (P = 0.0001) than the hip (P = 0.0004). Mild local skin irritation occurred in 12 subjects, resulting in discontinuation in only one subject. Except for the anticipated increase in hematocrit and hemoglobin, there were no clinically significant changes in blood counts or biochemistry. In three subjects with elevated serum prostate-specific antigen, prostate biopsies showed cancer. We conclude that continued application of AndroGel resulted in beneficial effects similar to those with injectables and other transdermal preparations. This study was neither placebo controlled nor powered to determine the effects of T treatment on prostate cancer risk. Thus, monitoring for prostatic disease and assessment for erythrocytosis are strongly advised to reduce the risk of adverse events with T treatment of hypogonadal men. PMID- 15126526 TI - Effect of levothyroxine replacement on lipid profile and intima-media thickness in subclinical hypothyroidism: a double-blind, placebo- controlled study. AB - Subclinical hypothyroidism (sHT) is associated with dyslipidemia and enhanced cardiovascular risk. We assessed carotid artery intima-media thickness (IMT, high resolution ultrasonography) and lipoprotein profile in 45 sHT patients (aged 37 +/- 11 yr) at baseline and after 6 months of randomized, placebo-controlled L T(4) replacement. In comparison with 32 age- and sex-matched controls, sHT patients had elevated total and low-density lipoprotein (LDL) cholesterol and ApoB levels (P = 0.002, P = 0.0007, and P = 0.01, respectively) and higher mean IMT values (P < 0.0001). In stepwise regression analysis, mean-IMT was positively related (r(2) = 0.71, P < 0.0001) to age, TSH, and LDL cholesterol. L-T(4) replacement significantly reduced both total and LDL cholesterol (P < 0.0001 for both) and mean-IMT (by 11%, P < 0.0001). The decrement in IMT was directly related to the decrements of both total cholesterol and TSH (P = 0.02 and P = 0.0001, respectively). We conclude that early carotid artery wall alterations are present in sHT patients. Whether such IMT increase is related to an early atherosclerotic involvement of the arterial wall cannot be clearly decided on the basis of the present results. However, the fact that L-T(4) replacement therapy was able to improve both the atherogenic lipoprotein profile and intima-media thickening suggests that lipid infiltration of arterial wall may represent a major mechanism underlying IMT increase in subclinical hypothyroidism. PMID- 15126527 TI - Activating Gsalpha mutations: analysis of 113 patients with signs of McCune Albright syndrome--a European Collaborative Study. AB - McCune-Albright syndrome (MAS) is a sporadic disorder characterized by the classic triad of polyostotic fibrous dysplasia, cafe-au-lait skin pigmentation, and peripheral precocious puberty. It is due to postzygotic activating mutations of arginine 201 in the guanine-nucleotide-binding protein (G protein) alpha subunit (Gsalpha), leading to a mosaic distribution of cells bearing constitutively active adenylate cyclase. MAS is heterogeneous: beyond the classic triad, a number of atypical or partial presentations have been reported. We present here the results of a systematic search for Gsalpha mutations in patients presenting with at least one of the signs of MAS, using a PCR-based sensitive method. We studied 113 patients (98 girls and 15 boys), 24% presenting the classic triad, 33% with two signs, and 40% with only one classic sign. Overall, the mutation was identified in 43% of the patients. When an affected tissue was available, the mutation was found in more than 90% of the patients, whatever the number of signs. Skin was a noteworthy exception because only three of the 11 skin samples were positive. The mutation was detected in 46% of blood samples in patients presenting the classic triad, whereas this figure fell to 21% and 8% in patients with two and one sign, respectively. Our results highlight the frequency of partial forms of MAS and the usefulness of sensitive techniques to confirm the diagnosis at the molecular level. It should be emphasized that we found the mutation in 33% of the 39 cases of isolated peripheral precocious puberty. This study has further widened the definition of MAS. Affections as clinically different as monostotic fibrous dysplasia, isolated peripheral precocious puberty, neonatal liver cholestasis, and the classic MAS all appear to be components of a wide spectrum of diseases based on the same molecular defect. PMID- 15126528 TI - Metabolic cardiovascular disease risk factors in women with self-reported symptoms of oligomenorrhea and/or hirsutism: Northern Finland Birth Cohort 1966 Study. AB - The metabolic cardiovascular disease (CVD) risk factors of women with self reported oligomenorrhea and/or hirsutism, which are symptoms of polycystic ovary syndrome (PCOS), were investigated in a general population-based Northern Finland Birth Cohort 1966 Study to determine whether women with PCOS symptoms at 31 yr would be distinguishable from asymptomatic controls in terms of CVD risk factors. A total of 518 cases with oligomenorrhea and/or hirsutism and 1036 randomly selected controls were analyzed. C-Reactive protein (CRP; median, 0.70 vs. 0.60 mg/liter; P = 0.026), triglycerides (mean, 0.97 vs. 0.91 mmol/liter; P = 0.039), body mass index (BMI; mean, 25.1 vs. 24.2 kg/m(2); P < 0.001), and waist/hip ratio (mean, 0.82 vs. 0.81; P = 0.001) were significantly higher, and high density lipoprotein cholesterol levels were lower (mean, 1.60 vs. 1.66 mmol/liter; P = 0.002) in the cases compared with the controls. Total cholesterol, low-density lipoprotein cholesterol, and blood pressure showed no statistically significant differences between the cases and the controls. In terms of metabolic CVD risk factors, women reporting hirsutism alone were indistinguishable from the control group, and those who reported both oligomenorrhea and hirsutism had the most severe changes in risk factor profiles. Because obesity is strongly related to PCOS symptoms, the analyses were stratified by BMI. After stratification into normal weight (BMI, <25 kg/m(2)), overweight (25 kg/m(2) or=30 kg/m(2)) groups, the waist/hip ratio was significantly higher among the overweight cases (mean, 0.84 vs. 0.83; P = 0.04). Among the obese women, high-density lipoprotein cholesterol was significantly lower (mean, 1.32 vs. 1.48 mmol/liter; P = 0.002) among the cases, and triglycerides tended to be higher (mean, 1.43 vs. 1.27 mmol/liter; P = 0.068) than in controls. In conclusion, these results indicate that self-reported symptoms of oligomenorrhea and/or hirsutism, particularly in the presence of both symptoms, may be helpful to identify women with metabolic cardiovascular risk factor accumulation associated with PCOS. PMID- 15126529 TI - Adverse effects of modest sleep restriction on sleepiness, performance, and inflammatory cytokines. AB - Total sleep restriction in humans is associated with increased daytime sleepiness, decreased performance, and hormonal/metabolic disturbances. The effects of mild chronic sleep restriction that mimic real life are not known. To assess the effects of modest sleep restriction from 8 to 6 h/night for 1 wk, 25 young, healthy, normal sleepers (12 men and 13 women) were studied for 12 consecutive nights in the sleep laboratory. After 1 wk of sleep restriction, although subjects' nighttime sleep was deeper, subjects were significantly sleepier (multiple sleep latency test) and performed worse in four primary variables of psychomotor vigilance test (both P < 0.01). Furthermore, 24-h secretion of IL-6 was increased by 0.8 +/- 0.3 pg/ml (P < 0.05) in both sexes, whereas TNFalpha was increased only in men. Also, the peak cortisol secretion was lower after sleep restriction than at baseline, and this difference was stronger in men (55.18 +/- 24.83 nmol/liter; P < 0.05) than in women (35.87 +/- 24.83 nmol/liter; P < 0.16). We conclude that in young men and women, modest sleep loss is associated with significant sleepiness, impairment of psychomotor performance, and increased secretion of proinflammatory cytokines. Given the potential association of these behavioral and physical alterations with health, well-being, and public safety, the idea that sleep or parts of it are optional should be regarded with caution. PMID- 15126530 TI - Imatinib mesylate (gleevec; STI571) monotherapy is ineffective in suppressing human anaplastic thyroid carcinoma cell growth in vitro. AB - Imatinib mesylate is remarkably effective in treating chronic myeloid leukemia and metastatic gastrointestinal stromal tumors. Meanwhile, anaplastic thyroid carcinoma (ATC) remains a fatal malignancy for which there are currently no effective curative interventions. In chronic myeloid leukemia and gastrointestinal stromal tumors, imatinib inhibits the constitutive tyrosine kinase activity of BCR-ABL and c-KIT, respectively. Reports suggest that imatinib may also be effective against ABL and platelet-derived growth factor receptor kinase-dependent pathological conditions. These mechanisms provide a wide scope of possible clinical applications for the drug. Potentially, diseases instigated by constitutive kinase activity that can be inhibited with imatinib should be treatable with this drug. We evaluated the effects of imatinib on the viability, cycling, and tyrosine phosphorylation of ATC cells in vitro. Our data indicate that imatinib has negligible antineoplastic activity against ATC cell lines within established therapeutically useful concentrations. No constitutive kinase activity was detected in these cell lines that could be exploited as a therapeutic target by imatinib. We conclude that imatinib mesylate monotherapy would not be effective in ATC patients. Current preclinical data do not warrant future clinical studies of imatinib monotherapy for ATC. PMID- 15126531 TI - Plasma ghrelin concentrations, food intake, and anorexia in liver failure. AB - Ghrelin is related to feeding behavior and nutrition in several physiological and pathological conditions. We tested the hypothesis that the anorexia and the decreased food intake of advanced liver failure might be associated with hyperghrelinemia. Fasting ghrelin was measured in 43 cirrhotic patients, food intake was self-assessed using the Corli score and a 3-d dietary record (n = 25), and anorexia/hunger was tested by a Likert scale. Fifty healthy subjects, matched for age and body mass index, served as controls. Ghrelin levels were not systematically increased in cirrhosis (414 +/- 164 vs. 398 +/- 142 pmol/liter in controls) but increased with decreasing Corli score (P = 0.014) and along the scale of anorexia/hunger (P = 0.0001), which were both related to the 3-d dietary record (P = 0.009 and P < 0.0001, respectively). Logistical regression confirmed that high ghrelin (>500 pmol/liter) was significantly associated with a low calorie intake [odds ratio (OR), 3.03 for any 100-calorie reduced intake; P = 0.015], a reduced Corli score (OR, 3.09; P = 0.031), and the anorexia score (OR, 3.37; P = 0.009), after adjustment for body mass index. The study confirms the previously observed relationship of fasting ghrelin with food intake in disease associated malnutrition. In the presence of anorexia, hyperghrelinemia might indicate a compensatory mechanism trying to stimulate food intake, which is nonetheless ineffective in the physiological range. PMID- 15126533 TI - Thyroid function and carotid wall thickness. AB - Decreased serum TSH levels predict vascular mortality in older people. There is a need to investigate mechanisms that could explain this association. This study was designed to investigate the relationship between thyroid function and the carotid intima-media thickness (IMT). The Study of Health in Pomerania is a population-based survey in Germany. Data from 2086 individuals at least 45 yr old with carotid ultrasound and without known thyroid disorders were analyzed. Twenty nine participants (1.4%) had elevated serum TSH levels, 300 (14.4%) had decreased serum TSH levels, and 12 (0.6%) participants were hyperthyroid. A linear relationship between thyroid function and IMT was found. The highest IMT values were observed in participants with hyperthyroidism, the lowest in subjects with elevated serum TSH levels (P < 0.01). A multivariable regression analysis identified thyroid function as an independent risk factor for increased IMT. Other risk factors for increased IMT included male gender, advanced age, diabetes mellitus, current smoking, and the use of antihypertensive medication; increased pulse pressure, serum low-density cholesterol, and total cholesterol/high-density lipoprotein ratio; as well as a decreased heart rate and a positive history of myocardial infarction. We conclude that there is an independent association between thyroid function and the IMT of the carotid artery. PMID- 15126532 TI - Rapid preoperative preparation for severe hyperthyroid Graves' disease. AB - Thyroidectomy (TX) is no longer the preferred choice for the therapy of hyperthyroid Graves' disease but is an alternative in patients who are noncompliant with or have reactions to antithyroid drugs, have moderate to severe ophthalmopathy, have large goiters, or who refuse (131)I therapy and/or long-term antithyroid drug therapy. Seventeen clinically and biochemically severely thyrotoxic patients (16 female, mean age of 35 yr), all but one with large goiters, underwent TX after rapid preparation. The potent inhibitors of the deiodination of T(4) to T(3), iopanoic acid (IOP) (500 mg twice a day) and dexamethasone (DEX) (1 mg twice a day), were given with propylthiouracil or methimazole, when possible, and beta-blockers. Thyroid function tests were obtained before treatment and at TX. All patients were thyrotoxic (mean +/- SE: T(4), 21.6 +/- 1.2 micro g/dl; free T(4) index (FTI), 10.3 +/- 0.8; total T(3), 510 +/- 48 ng/dl). IOP and DEX rapidly lowered T(3) values (P < 0.0001; total T(3), 147 +/- 13 ng/dl) with a smaller but significant (P < 0.05) decrease in T(4)/FTI (T(4), 17.9 +/- 1.3 micro g/dl; FTI, 7.9 +/- 0.6). All patients were clinically euthyroid before surgery. None developed hypoparathyroidism, laryngeal nerve damage, or worsening of ophthalmopathy after surgery. The restoration of hyperthyroid Graves' disease to euthyroidism is rapidly accomplished with IOP and DEX, beta-blockers, and, when possible, antithyroid drugs. This is especially relevant in noncompliant patients with large goiters. PMID- 15126534 TI - Autosomal-dominant pseudohypoaldosteronism type 1 in a Turkish family is associated with a novel nonsense mutation in the human mineralocorticoid receptor gene. AB - Pseudohypoaldosteronism type 1 (PHA1) is a rare congenital disease inherited in either an autosomal-recessive or an autosomal-dominant trait. The autosomal dominant form manifests with renal salt loss in infancy and a gradual improvement with advancing age. PHA1 presents with potential life-threatening salt wasting and failure to thrive in early infancy. Autosomal-dominant forms of PHA1 are often caused by heterozygous mutations of the MR gene coding for the mineralocorticoid receptor. Whether heterozygous mutations of the MR gene impair biological function as a result of haplo-insufficiency or due to a dominant negative effect needs further clarification. We report a case of a renal form of PHA1 in a Turkish family. A heterozygous nonsense mutation c3055C>T (R947X) in exon 9 of the MR gene leading to a premature stop codon was identified in the index patient. The truncated receptor is free of aldosterone binding. The segregation analysis revealed the identical mutation in the patient's father, who never showed any symptoms of PHA. This shows the incomplete penetrance of the phenotype, although a mild salt loss might have been overlooked in the father's childhood. PMID- 15126535 TI - Exogenous and endogenous postprandial lipid abnormalities in type 2 diabetic patients with optimal blood glucose control and optimal fasting triglyceride levels. AB - The aim of this study was to evaluate exogenous and endogenous lipoprotein responses to a standard fat-rich meal in type 2 diabetic patients with optimal fasting triglyceridemia and optimal blood glucose control. Seven type 2 diabetic patients and five nondiabetic controls (age, 49 +/- 7 and 48 +/- 4 yr; body mass index, 28.3 +/- 3.6 and 25.1 +/- 3.6 kg/m(2); mean +/- SD) were given, after at least 12 h of fasting, a standard fat-rich meal. Before and over the 6 h after the meal, serial blood samples were taken for determination of glucose, insulin, lipids, lipoproteins, apolipoprotein B-48 (apo B-48), apo B-100, free fatty acids, and lipoprotein lipase activity. The main abnormality in the postprandial lipid response of diabetic patients involved large very low density lipoproteins. In these particles, apo B-48, apo B-100, cholesterol, and triglyceride incremental areas were, in fact, significantly higher in diabetics compared with controls [7.08 +/- 2.65 vs. 1.17 +/- 0.88 mg/liter.h, 65.5 +/- 11.5 vs. 12.4 +/- 1.77 mg/liter.h, 29.7 +/- 3.9 vs. 13.1 +/- 3.1 mg/dl.h (0.77 +/- 0.10 vs. 0.34 +/ 0.08 mmol/liter.h), 170 +/- 31 vs. 94 +/- 22 mg/dl.h (1.93 +/- 0.35 vs. 1.06 +/- 0.25 mmol/liter.h)] (all P < 0.05; mean +/- SEM). Postprandial preheparin lipoprotein lipase plasma activity was, if anything, higher in diabetic patients. In conclusion, even with fasting normotriglyceridemia and optimal blood glucose control, type 2 diabetic patients are characterized, in the postprandial period, by a significant increase in large very low density lipoproteins of both endogenous and exogenous origins. PMID- 15126536 TI - Increased C-reactive protein levels in the polycystic ovary syndrome: a marker of cardiovascular disease. AB - The polycystic ovary syndrome (PCOS), one of the most common reproductive abnormalities, shares some components of the metabolic cardiovascular syndrome. Therefore, PCOS patients may represent the largest group of women at high risk for the development of early-onset cardiovascular disease (CVD) and/or diabetes. C-reactive protein (CRP) is a strong independent predictor of future CVD and/or stroke. Only one small published study has looked for such an association (17 PCOS patients vs. 15 controls). The objective of this study was to compare the levels of CRP and other risk factors of CVD in a large group of PCOS patients and controls. CRP measurements were undertaken in 116 PCOS patients and 94 body mass index-matched controls with regular menstrual cycles. Whereas 36.8% of the PCOS patients had CRP levels above 5 mg/liter, only 9.6% of the controls exhibited high CRP levels (P < 0.001). The mean +/- SD was 5.46 +/- 7.0 in the PCOS group vs. 2.04 +/- 1.9 mg/liter in the control (P < 0.001). The body mass index, white blood cell count, TSH, glucose, cholesterol, and homocysteine levels were not significantly different between the two groups. CRP levels are elevated in patients with PCOS and may be a marker of early cardiovascular risk in these patients. High CRP levels may explain why some PCOS women may possibly be at an increased risk for the development of early-onset CVD. Consequently, whether treatment regimens directed toward lowering CVD risk factors should be more aggressive for those PCOS women with increased CRP levels, awaits further clinical experience. PMID- 15126537 TI - High habitual calcium intake attenuates bone loss in early postmenopausal Chinese women: an 18-month follow-up study. AB - This study assessed the association of habitual dietary calcium intake and bone loss in early postmenopausal women. Four hundred fifty-four healthy postmenopausal Chinese women were enrolled for this 18-month cohort study. The subjects were 48-62 yr of age and within 12 yr of natural menopause. Dietary intake was assessed by the food frequency method, and bone mass was measured using dual energy x-ray absorptiometry at baseline and 9 and 18 months. The association between mean habitual dietary intake over the follow-up period and the rate of bone loss was examined. During the 18-month follow-up, the total loss rates of BMD at the whole body, lumber spine, femoral neck, and total hip were 1.28, 0.60, 1.54, and 0.56% (all P < 0.01). Subjects were stratified into four quartiles according to calcium intake during the period of follow-up. Quartiles I IV had median intakes of 341, 505, 682, and 934 mg Ca/d. Subjects in quartile IV had significantly less BMD loss at the whole body and less BMD/bone mineral content loss at Ward's triangle, even after adjustments for confounding factors (by analysis of covariance). Multiple linear regression analyses showed significant positive associations between calcium intake and BMD change at the whole body (P = 0.006) and Ward's triangle (P = 0.021). Calcium intake was significantly associated with bone mineral content change at the trochanter (P = 0.025) and Ward's triangle (P < 0.001). No significant effect of calcium intake at the spine was found. In conclusion, habitual dietary calcium intake had a beneficial effect on bone loss at the whole body and some regions of the hip. Our findings suggest that an intake exceeding 900 mg calcium/d was helpful in the prevention of cortical bone loss among early postmenopausal Chinese women. PMID- 15126538 TI - Differential effects of metformin and exercise on muscle adiposity and metabolic indices in human immunodeficiency virus-infected patients. AB - The HIV-lipodystrophy syndrome is associated with fat redistribution and metabolic abnormalities, including insulin resistance (IR). The mechanisms and treatment strategies for IR in HIV-lipodystrophy are unclear, but data suggest that intramuscular lipids contribute to IR in this population. We previously showed that metformin and exercise improve hyperinsulinemia more than metformin alone in HIV-lipodystrophy. Now we investigate the effects of these treatment strategies on thigh muscle adiposity measured by computed tomography and additional body composition measures. Twenty-five HIV-infected patients on stable antiretroviral therapy with hyperinsulinemia and fat redistribution participated in a prospective, randomized, 3-month study of metformin alone or metformin and resistance training three times a week. Thigh muscle adiposity decreased significantly more as shown by increased muscle attenuation [2.0 (range, 0.5-5.0) vs. -1.0 (-3.5-0), P = 0.04] and sc leg fat tended to decrease more [-3.3 (-7.5 4.3) vs. 0.8 (-2.1-9.5), P = 0.06] in the combined treatment group in comparison with metformin alone. In multivariate analysis, change in thigh muscle adiposity remained a significant predictor of change in insulin (P = 0.04), controlling for changes in other body composition measurements. These data suggest that muscle adiposity, in addition to other fat depots, is an important determinant of hyperinsulinemia and that exercise has complex effects on regional fat depots in HIV-infected patients. Reduction in muscle adiposity may be an important mechanism by which exercise improves hyperinsulinemia in this population. PMID- 15126539 TI - Marked suppression of dihydrotestosterone in men with benign prostatic hyperplasia by dutasteride, a dual 5alpha-reductase inhibitor. AB - Dihydrotestosterone (DHT) is the primary metabolite of testosterone in the prostate and skin. Testosterone is converted to DHT by 5alpha-reductase, which exists in two isoenzyme forms (types 1 and 2). DHT is associated with development of benign prostatic hyperplasia (BPH), and reduction in its level with 5alpha reductase inhibitors improves the symptoms associated with BPH and reduces the risk of acute urinary retention and prostate surgery. A selective inhibitor of the type 2 isoenzyme (finasteride) has been shown to decrease serum DHT by about 70%. We hypothesized that inhibition of both isoenzymes with the dual inhibitor dutasteride would more effectively suppress serum DHT levels than selective inhibition of only the type 2 isoenzyme. A total of 399 patients with BPH were randomized to receive once-daily dosing for 24 wk of dutasteride (0.01, 0.05, 0.5, 2.5, or 5.0 mg), 5 mg finasteride, or placebo. The mean percent decrease in DHT was 98.4 +/- 1.2% with 5.0 mg dutasteride and 94.7 +/- 3.3% with 0.5 mg dutasteride, significantly lower (P < 0.001) and with less variability than the 70.8 +/- 18.3% suppression observed with 5 mg finasteride. Mean testosterone levels increased but remained in the normal range for all treatment groups. Dutasteride appeared to be well tolerated with an adverse event profile similar to placebo. PMID- 15126540 TI - Familial concordance of thyroid and other head and neck tumors in an irradiated cohort: analysis of contributing factors. AB - Relatively little is known about variations in susceptibility to the effects of radiation in the general population. We have been studying 4296 individuals exposed as children to head and neck radiation. The present study was designed to evaluate the pattern of thyroid, parathyroid, salivary, and neural tumors in irradiated siblings for evidence of heritable susceptibility factors. We also wanted to determine whether the characteristics of thyroid cancers were influenced by familial factors. The following criteria were met by 251 sibling pairs: both irradiated, both with follow-up (average, 44.3 +/- 9.4 yr; range, 9.4 59.5 yr), and both with organ-dose estimates. For each sibling pair we derived a quantitative score, taking into account the length of follow-up and known risk factors, for their concordance and used the sum of these scores to characterize the population. Whether we used thyroid cancer or all thyroid nodules as an end point, the degree of concordance did not exceed what could be explained by the length of follow-up and known risk factors. For thyroid cancer, neither the presenting characteristics nor their rates of recurrence were influenced by their concordance status. In summary, we were unable to identify familial factors that modify the strong effects of radiation exposure. There is no reason to alter the evaluation or treatment of thyroid cancer in an irradiated patient based on whether another member of the family has radiation-related tumors. PMID- 15126541 TI - Impact of growth hormone (GH) treatment on cardiovascular risk factors in GH deficient adults: a Metaanalysis of Blinded, Randomized, Placebo-Controlled Trials. AB - Patients with hypopituitarism have an increased risk of cardiovascular mortality. GH treatment could modify the cardiovascular risk in adults with GH deficiency, but most published clinical trials involved few patients and the results are variable. We conducted a systematic review of blinded, randomized, placebo controlled trials of GH treatment in adult patients with GH deficiency published up to August 2003. Thirty-seven trials were identified. We combined the results for effects on lean and fat body mass; body mass index; triglyceride and cholesterol [high-density lipoprotein, low-density lipoprotein (LDL), and total] levels; blood pressure; glycemia; and insulinemia. Overall effect size was used to evaluate significance, and weighted differences between GH and placebo were used to appreciate the size of the effect. GH treatment significantly reduced LDL cholesterol [-0.5 (SD 0.3) mmol/liter], total cholesterol [-0.3 (0.3) mmol/liter], fat mass [-3.1 (3.3) kg], and diastolic blood pressure [-1.8 (3.8) mm Hg] and significantly increased lean body mass [+2.7 (2.6) kg], fasting plasma glucose [+0.2 (0.1) mmol/liter], and insulin [+8.7 (7.0) pmol/liter]. All effect sizes remained significant in trials with low doses and long-duration GH treatment. Thus, GH treatment has beneficial effects on lean and fat body mass, total and LDL cholesterol levels, and diastolic blood pressure but reduces insulin sensitivity. The global cardiovascular benefit remains to be determined in large trials with appropriate clinical endpoints. PMID- 15126542 TI - Pituitary magnetic resonance imaging in 15 patients with Prop1 gene mutations: pituitary enlargement may originate from the intermediate lobe. AB - Pituitary morphology in patients with Prop1 gene mutations varies. Most patients demonstrate a normal or small pituitary gland. Occasionally, pituitary enlargement of undetermined origin has also been detected. In the present study we use long-term magnetic resonance imaging findings to characterize the morphological abnormalities of the pituitary gland in 15 patients (aged 2.5-45 yr) with combined pituitary hormone deficiency caused by Prop1 gene mutations (GA296del/GA296del in seven, GA296del/A150del in two, A150del/A150del in five, and GA296del/R73H in one patient) and attempt to uncover the origin and nature of the pituitary enlargement. Small pituitary gland was detected in seven patients (25.2 +/- 14.4 yr of age), normal pituitary size in three patients (10.2 +/- 5.8 yr of age), and pituitary enlargement in five patients (6.5 +/- 2.7 yr of age). The pituitary enlargement consisted of a nonenhancing mass lesion interposed between the normally enhancing anterior lobe and the neurohypophysis. The pituitary stalk was displaced anteriorly, whereas the neurohypophysis was orthotopic, displaying a normal signal. Spontaneous regression of the mass lesion with normalization of the pituitary stalk position was observed in three patients. Our data indicate that although a small pituitary gland is usually observed in older subjects, a significant number of young patients with Prop1 gene mutations demonstrate pituitary enlargement with subsequent regression. The distinct magnetic resonance imaging characteristics of the pituitary enlargement in our patients in conjunction with pertinent data from Prop1-deficient mice suggest that the mass causing the pituitary enlargement most likely originates from the intermediate lobe. PMID- 15126543 TI - Effect of the combination of methyltestosterone and esterified estrogens compared with esterified estrogens alone on apolipoprotein CIII and other apolipoproteins in very low density, low density, and high density lipoproteins in surgically postmenopausal women. AB - Androgens are known to lower plasma triglycerides, an independent risk factor for coronary heart disease (CHD). Triglycerides are carried in plasma on very low density (VLDL) and low density (LDL) lipoprotein particles. Apolipoprotein CIII (apoCIII), a strong predictor of CHD, impairs the metabolism of VLDL and LDL, contributing to increased triglycerides. The objective of this study was to assess the effect of oral methyltestosterone (2.5 mg/d), added to esterified estrogens (1.25 mg/d), on concentrations of apolipoproteins and lipoproteins, specifically those containing apoCIII, compared with esterified estrogens alone in surgically postmenopausal women. The women in the methyltestosterone plus esterified estrogen group had significant decreases in total triglycerides, apoCI, apoCII, apoCIII, apoE, and high density lipoprotein (HDL) cholesterol compared with those in the esterified estrogen group. The decreases in apoCIII concentrations occurred in VLDL (62%; P = 0.02), LDL (35%; P = 0.001), and HDL (17%; P < 0.0001). There were also decreases in cholesterol and triglycerides concentrations of apoCIII containing LDL, and apoCI concentration of apoCIII containing VLDL. There was no effect on VLDL and LDL particles that did not contain apoCIII or on apoB concentrations. In conclusion, methyltestosterone, when administered to surgically postmenopausal women taking esterified estrogen, has a selective effect to reduce the apoCIII concentration in VLDL and LDL, a predictor of CHD. Methyltestosterone may lower plasma triglycerides through a reduction in apoCIII. PMID- 15126544 TI - The role of [(18)F]fluorodeoxyglucose positron emission tomography and [(111)In] diethylenetriaminepentaacetate-D-Phe-pentetreotide scintigraphy in the localization of ectopic adrenocorticotropin-secreting tumors causing Cushing's syndrome. AB - Conventional imaging modalities cannot localize the source of ACTH in 30-50% of patients with Cushing's syndrome (CS) caused by ectopic ACTH secretion (EAS). We prospectively evaluated whether [(18)F]fluorodeoxyglucose (FDG) positron emission tomography (PET) or [(111)In]-diethylenetriaminepentaacetate-D-Phe-pentetreotide (OCT) at higher than standard doses of radionuclide (18 mCi; H-OCT), can detect these tumors. Seventeen patients with presumed EAS based on inferior petrosal sinus sampling results underwent routine anatomical imaging studies [computed tomography (CT) and magnetic resonance imaging (MRI)] and OCT scintigraphy with 6 mCi (L-OCT). Research studies included FDG-PET in all patients and H-OCT if L-OCT was negative. ACTH-secreting tumors were localized in 13 patients and were occult in four. Nine of 17 CT, six of 16 MRI, six of 17 FDG-PET, eight of 17 L-OCT, and one of nine H-OCT studies were true positives. The sensitivity of CT and combined H- and L-OCT scintigraphy was higher (both 53%; 95% confidence interval, 29-76%) than that of MRI (37%; 95% confidence interval, 16-64%) or FDG-PET (35%; 95% confidence interval, 15-61%). FDG-PET did not detect tumors that were occult on CT/MRI. L-OCT was a useful complementary modality to CT and MRI. As H-OCT identified a tumor in one patient with otherwise negative imaging, it should be considered only when other imaging modalities fail to localize the ACTH-secreting tumor in patients with EAS. PMID- 15126545 TI - Autoimmunity to islet proteins in children diagnosed with new-onset diabetes. AB - Autoantibody and T cells reacting with islet proteins have been demonstrated in patients with type 1 diabetes. In recent years an increasing number of children have been clinically classified with type 2 (not ketosis prone, evidence of insulin resistance, presence of acanthosis nigricans, and obesity) or indeterminant diabetes (admixture of clinical features of types 1 and 2). In this study, we compared the islet cell autoantibody and T-cell responses to islet proteins in type 2 (n = 19) and indeterminant (n = 16) children (<18 yr of age) to classic type 1 (n = 37) diabetic patients. We observed that 37 of 37 type 1 diabetic children demonstrated autoantibody and/or T-cell reactivity to islet proteins. Fourteen of the 19 type 2 patients were positive for islet cell autoantibodies, and six of 14 were positive for T-cell responses to islet proteins. For the indeterminant patients, 11 of 16 of the patients were positive for autoantibodies, and six of 16 patients were positive for T-cell responses to islet proteins. These results demonstrate that autoimmunity to islet proteins is present in a high percentage of children classified as indeterminant or type 2 diabetes. Moreover, the presence of obesity or acanthosis nigracans does not reliably distinguish children with or without islet cell autoimmunity. PMID- 15126546 TI - Endocrinologic and psychologic evaluation of 21-hydroxylase deficiency carriers and matched normal subjects: evidence for physical and/or psychologic vulnerability to stress. AB - Carriers of congenital adrenal hyperplasia due to 21-hydroxylase (21-OH) deficiency demonstrate increased secretion of cortisol precursors after ACTH stimulation, suggestive of impaired cortisol production and compensatory increases in hypothalamic CRH secretion. Because both cortisol and CRH have behavioral effects, and hypothalamic CRH hypersecretion has been associated with chronic states of anxiety and depression, we performed endocrine and psychologic studies in consecutively admitted parents of patients with classic congenital adrenal hyperplasia due to 21-OH deficiency and parents of children with other chronic endocrine disorders. The number of excluded carriers because of pathologic reasons was higher than that of controls (P = 0.05). Carriers of 21-OH deficiency had a lower mean 24-h urinary free cortisol excretion (26.4 +/- 3.4 vs. 42.7 +/- 6.4 microg/d, P = 0.03) and higher peak ACTH (75.7 +/- 8.1 vs. 54.2 +/- 5.9 pg/ml, P = 0.04) and 17-hydroxyprogesterone (224.2 +/- 28.1 vs. 107.1 +/- 12.5 ng/dl, P < 0.001) concentrations post CRH stimulation than control subjects. Cortisol and androstenedione responses were similar in the two groups. Psychometric assessment performed by administering the State-Anxiety Inventory, Beck Depression Inventory, Profile of Mood States, Symptom Checklist-90R, and Temperament and Character Inventory revealed no differences between the two subject groups. Interestingly, a stepwise multiple linear regression model analysis in each population sample revealed that in carriers of 21-OH deficiency but not in the control subjects, a lower mean 24-h urinary free cortisol excretion and a higher ACTH response to ovine CRH stimulation predicted predisposition to obsessive-compulsive behavior, novelty seeking, reward dependence, and harm avoidance. We conclude that carriers of 21-OH deficiency appear to have mild hypocortisolism and compensatory changes of CRH secretion secondary to lower cortisol concentrations. These changes might predict mild predisposition of these subjects to physical and psychologic pathology, suggesting that larger studies are necessary. PMID- 15126547 TI - Spinal volumetric bone mineral density and vertebral fractures in female patients with adrenal incidentalomas: the effects of subclinical hypercortisolism and gonadal status. AB - Although adrenal incidentalomas (AI) are not associated with clinically evident syndromes, some patients display biochemical features of subclinical hypercortisolism (SH). Previous studies indicated a negative effect of SH on bone in AI patients, but the prevalence of vertebral fractures and the roles of SH and gonadal status in volumetric bone mineral density are unknown. In 70 female AI patients and 84 controls, the prevalence of vertebral fractures and spinal bone mineral density (by quantitative computed tomography) were evaluated. Subjects were subdivided according to menopausal status into groups Pre (21 patients and 23 controls) and Post (49 patients and 61 controls); there were 14 and 35 patients without SH (SH(-)) and 7 and 14 patients with SH (SH(+)) in groups Pre and Post, respectively. The prevalence of fractures was higher in SH(+) than in controls and in SH(-) subjects in both groups Pre [SH(+), 42.9%; controls, 0% (P = 0.001); SH(-), 7.1% (P = 0.049)] and post [SH(+), 78.6%; controls, 37.7% (P = 0.006); SH(-) 42.9% (P = 0.024)]. In group Post, the mean z-score quantitative computed tomography values were lower in SH(+) patients (-0.78 +/- 0.29) than in controls (0.06 +/- 0.14; P = 0.011) and SH(-) patients (0.02 +/- 0.19; P = 0.034). Evaluation of spinal bone is indicated in female AI patients with SH. PMID- 15126548 TI - Effects of 0.9 mg recombinant human thyrotropin on thyroid size and function in normal subjects: a randomized, double-blind, cross-over trial. AB - The effect of recombinant human TSH (rhTSH) on thyroid function and ultrasonically determined thyroid volume was investigated in nine healthy euthyroid male volunteers. Each received either 0.9 mg rhTSH or isotonic saline in a randomized order, and thyroid volume and function were closely monitored during the following 28 d. No significant changes were observed after saline injection. After rhTSH stimulation, the median serum TSH increased from 2.03 mU/liter (range, 0.99-3.07 mU/liter) to more than 200 mU/liter (range, 78.9 to >200.0 mU/liter) after 4 h, with a subsequent rapid decline. Mean (+/-SEM) serum free T(4) and free T(3) peaked at 48 h with levels 204.7 +/- 26.1% and 226.9 +/- 31.4%, respectively, above baseline (P < 0.001). Twenty-four hours after rhTSH stimulation, mean (+/-SEM) thyroid volume was significantly increased by 23.3 +/- 5.8% (P = 0.003) and after 48 h by 35.5 +/- 18.4% (P = 0.02). On d 4 the mean thyroid enlargement had reverted to baseline values. One individual developed a 90-ml tender thyroid enlargement (initially 21 ml) 36 h after rhTSH administration, associated with a very high level of serum thyroglobulin. It is concluded that 0.9 mg rhTSH may result in a profound stimulation of not only thyroid function but also of thyroid size, appearing in the period 1-4 d after injection. Further dose-response studies are needed to clarify the potential hazards before routine use, for example in the context of (131)I therapy and goiter. PMID- 15126549 TI - Weight and body composition changes during and after adjuvant chemotherapy in women with breast cancer. AB - Uncontrolled trials have reported significant weight gain in women with breast cancer during treatment with adjuvant chemotherapy. We prospectively evaluated body composition before (visit 1), immediately after (visit 2), and 6 months after (visit 3) chemotherapy in 20 women with stages I-IIIA breast cancer [body mass index (BMI): 24.1 +/- 3.9 kg/m(2)]. We compared their weight change to 51 age- and BMI-matched healthy controls (BMI: 25.5 +/- 3.8 kg/m(2)). In women with breast cancer, there was no weight change from visit 1-2, or from visit 1-3, but weight increased from visit 2-3 (+1.09 +/- 2.46 kg; P = 0.05). Weight change was not different from controls during either interval. In the breast cancer group, the percentage of body fat assessed by air displacement plethysmography increased, and fat-free mass decreased from visit 1-2 (+2.3 +/- 4% and -2.2 +/- 4%; P = 0.02) and from visit 1-3 (+4.0 +/- 6% and -3.8 +/- 6%; P = 0.01). By dual energy x-ray absorptiometry, the percentage of body fat increased from visit 2-3 (+0.9 +/- 1.6%; P = 0.02). Bone mineral content decreased from visit 2-3 (-0.02 +/- 0.04 kg; P = 0.02) and from visit 1-3 (-0.04 +/- 0.06 kg; P = 0.005). By computed tomography, the visceral adipose to sc adipose tissue ratio decreased from visit 1-3 (-0.02 +/- 0.05 ml; P = 0.02). We conclude that, compared with controls, women with breast cancer receiving modern adjuvant chemotherapy regimens show no significant changes in weight during the first year of their treatment. They do, however, appear to undergo unfavorable changes in body composition. PMID- 15126550 TI - Male hormonal contraception: effects of injections of testosterone undecanoate and depot medroxyprogesterone acetate at eight-week intervals in chinese men. AB - Surveys indicate that one form of acceptable male hormonal contraception would consist of injections given at 2- to 3-month intervals. This report describes a study of depot medroxyprogesterone acetate (DMPA) and testosterone undecanoate (TU) injected at 8-wk intervals for suppression of spermatogenesis in healthy Chinese men. After screening, 30 healthy volunteers were enrolled and randomly assigned to one of three dose groups (n = 10/group): 1000 mg TU (group A); 1000 mg TU plus 150 mg DMPA (group B); 1000 mg TU plus 300 mg DMPA (group C). All doses were given as im injections at 8-wk intervals. The study consisted of an 8 wk control (baseline) period, a 24-wk treatment period, and a 24-wk recovery period. Consistent azoospermia or severe oligozoospermia was achieved and maintained in all volunteers during the treatment period, except for two men in the TU-alone group who experienced a rebound in sperm concentrations. An 8-wk regimen of TU plus DMPA at both tested combination doses effectively suppressed spermatogenesis to azoospermia in Chinese men. All volunteers tolerated the injections; no serious adverse effects were reported. The lower-dose combination is recommended for further testing in an expanded clinical trial or contraceptive efficacy study. PMID- 15126551 TI - Klinefelter syndrome in adolescence: onset of puberty is associated with accelerated germ cell depletion. AB - The process of germ cell depletion in patients with Klinefelter syndrome (KS) is incompletely characterized. In the current work, we evaluated the presence of germ cells in adolescent boys with KS for possible future use in assisted reproduction techniques. Fourteen nonmosaic 47,XXY boys (aged 10-14 yr) were enrolled. Every fourth month their puberty was staged, and serum was obtained for hormone analyses. Each boy underwent a single testicular biopsy. Biopsy specimens of seven peripubertal boys (testicular volume < 2.0 ml) had spermatogonia of adult type, whereas older boys with larger testes (> 2.0 ml) exhibited no germ cells. No meiotic germ cells were detectable in any of these subjects. Depletion of germ cells was associated with an increase in testicular volume but was not immediately reflected in levels of serum gonadotropin, inhibin B, or anti Mullerian hormone. In contrast, hypergonadotropism and suppression of serum inhibin B and anti-Mullerian hormone developed later, during midpuberty, after an unequivocal increase in serum testosterone (>2.5 nmol/liter) levels and degeneration of Sertoli cells. In conclusion, these prepubertal and early pubertal boys with KS had diploid germ cells that vanished in early puberty when testicular volume increased, whereas serum gonadotropin and inhibin B levels displayed pathological changes later during midpuberty. PMID- 15126552 TI - Variability and reliability of single serum IGF-I measurements: impact on determining predictability of risk ratios in disease development. AB - In recent years, a number of investigators have studied the relationship between IGF-I and risk of developing cancer, diabetes or cardiovascular disease. Upper tertile, quartile, and quintile IGF-Is were associated with higher risk of developing cancer, and lowest quartile with cardiac disease and diabetes. As part of a study to correlate serum IGF-Is and growth hormone dynamics in aging, we measured fasting serum IGF-I at baseline and two weeks later in a group of 84 normal volunteers between the ages of 50 and 90 years. Although the correlation between the two IGF-Is was high (r=0.922; p<0.0001) there were substantial differences between the two IGF-I values ranging from -36.25 to +38.24% between individual IGF-I values at the two blood draws and a significant difference between the mean IGF-Is at visits I and 2 (mean 120.28+/-53.5 vs. 114.95+/-50.03; p=0.03). When considered in quartiles, IGF-I changed from one quartile to another in 34/84 (40.5%) of the volunteers. When the group was divided in halves, tertiles,quartiles, or quintiles there was an increasing number of subjects who changed from one subdivision to another as the number of gradations increased. These results suggest that the predictive outcomes of earlier studies that used single IGF-I samples for analysis of risk ratios according to tertiles, quartiles, or quintiles could have been different if a second IGF-I was used to establish the risk ratio. The results also suggest that variability in IGF-I should be taken into account when designing such studies. PMID- 15126553 TI - The ontogeny of pulsatile growth hormone secretion and its temporal relationship to the onset of puberty in the agonadal male rhesus monkey (Macaca mulatta). AB - The pubertal amplification of GH secretion in primates has been thought to reflect an increase in gonadal steroid hormones due to gonadotropin stimulation induced by hypothalamic GnRH release. Previous studies in agonadal, peripubertal, male rhesus monkeys have estimated the age of GnRH activation (defined as d 0) using analyses of nocturnal, pulsatile LH patterns derived from sequential blood samples. Using samples from these earlier studies, secretory patterns of GH were analyzed using Cluster at approximately 30-d intervals in the youngest prepubertal ages and at approximately 10- to 20-d intervals in the period immediately preceding and following the onset of puberty. Pulse frequency, amplitude, and mean GH increased significantly between early prepubertal ages (up to 30 d before d 0) and the late prepubertal period (between -20 d and d 0). Pulsatile GH activity increased earlier than pulsatile LH secretion in four of five animals. These findings support the conclusion that pulsatile GH secretion increases developmentally in the absence of gonadal steroids. Furthermore, the present observation that the developmental increase in GH secretion occurs earlier than previously reported is consistent with the possibility that GH itself either directly or indirectly participates in the pubertal reinitiation of GnRH pulse generator activity. PMID- 15126554 TI - Ret/PTC activation in benign and malignant thyroid tumors arising in a population exposed to low-dose external-beam irradiation in childhood. AB - Ionizing radiation is the strongest risk factor known for the development of thyroid neoplasia. Although ret/PTC rearrangements have been identified in both spontaneous and radiation-induced papillary thyroid cancer, they seem more frequent among radiation-associated tumors. We studied the frequency of ret/PTC activation in a group of sporadic and radiation-induced thyroid carcinomas (n = 49) and adenomas (n = 13) among 44 individuals treated for Tinea Capitis with low dose external irradiation as well as in 18 nonirradiated subjects. Total RNA recovered from paraffin-embedded thyroid cancer surgical specimens was analyzed for ret/PTC 1, 2, and 3 mutations using RT-PCR with Southern blotting to maximize detection sensitivity. Ret/PTC rearrangements were identified in 42.9% of thyroid carcinoma and 46.2% of adenoma subjects. Among the positive carcinoma specimens, three were follicular carcinomas. Ret/PTC 1, the predominant rearrangement, was more prevalent in nonirradiated compared with irradiated carcinomas (66.7 vs. 27.0%; P = 0.04). Ret/PTC activation was associated with male gender. The strengths of this study included analysis of age-, gender-, and ethnicity-matched groups; molecular analysis using two techniques; and a complete blinding of laboratory analysis from clinical features. The differences seen between these and other published results may be related to differences in radiation doses to the thyroid, latency period between time of radiation exposure and development of clinically apparent thyroid cancer, and ethnic background of the study populations. PMID- 15126555 TI - Sustained elevation of pulsatile growth hormone (GH) secretion and insulin-like growth factor I (IGF-I), IGF-binding protein-3 (IGFBP-3), and IGFBP-5 concentrations during 30-day continuous subcutaneous infusion of GH-releasing peptide-2 in older men and women. AB - We test the interlinked hypotheses that in healthy older adults: 1). i.v. injection of GH-releasing peptide-2 (GHRP-2) and GHRH synergizes more in aging women than men; 2). sc infusion of both GHRP-2 (1 microg/kg.h = 1) and GHRH (1, 3, or 10) for 24 h augments GH secretion more than either agonist alone; and 3). continuous sc delivery of GHRP-2 (1) for 30 d stimulates daily GH secretion and IGF-I, IGF-binding protein-3 (IGFBP-3), and IGFBP-5. Acute two-peptide synergy was 3-fold greater in young (n = 16) than older volunteers (n = 17; P < 0.025) and was 2.3-fold higher in elderly women than men (P < 0.025). The 24-h infusion of GHRP-2 (1) combined with GHRH (3 or 10) in men and with GHRH (10) in women drove GH secretion more than GHRH alone (P 4 ng/mL, 5th versus 1st quintile OR = 0.49, 95% CI = 0.28 to 0.86; P(trend) =.002). These inverse associations were observed in both pre- and post-PSA eras. CONCLUSIONS: The inverse association between baseline plasma selenium levels and risk of advanced prostate cancer, even among men diagnosed during the post-PSA era, suggests that higher levels of selenium may slow prostate cancer tumor progression. Ongoing randomized trials of selenium supplements may help to further evaluate this issue. PMID- 15126607 TI - Lymphatic drainage imaging of breast cancer in mice by micro-magnetic resonance lymphangiography using a nano-size paramagnetic contrast agent. AB - BACKGROUND: The presence of lymph node metastases is an important factor in breast cancer patient prognosis. Therefore, the precise identification of sentinel lymph nodes in these patients is critical. Improving current magnetic resonance (MR) imaging methods using a newly synthesized nano-size paramagnetic molecule, G6, as a contrast agent, provides an attractive means toward attaining this goal. METHODS: A four-dimensional method of micro-MR lymphangiography using G6 (9 nm/240 kd) was developed to visualize the lymphatic ducts and lymph nodes draining mouse mammary tumors over time. The ability of micro-MR lymphangiography with the G6 contrast agent to visualize lymphatic drainage of normal mouse mammary tissue was compared with that of the conventional MR contrast agent, Gd [DTPA]-dimeglumine (<1 kd). Lymphatic drainage in spontaneous and xenografted breast tumor models was visualized using the G6 contrast agent. RESULTS: Draining lymphatic ducts and lymph nodes were clearly visualized in the mammary tissue of normal mice and in spontaneous and xenografted breast tumor models after a direct mammary gland or peritumoral injection of G6. Gd-[DTPA]-dime-glumine, by contrast, failed to depict lymphatic flow from the mammary tissue in normal mice using the same method. Micro-MR lymphangiography using the G6 contrast agent revealed the absence of filling in the metastatic foci of affected lymph nodes. CONCLUSIONS: The superior temporal and spatial resolution of micro-MR lymphangiography using the contrast agent G6 may facilitate the study of tumor lymphatic drainage and lymphatic metastasis in both experimental animals and clinical medicine. In addition, this may be a powerful new method for sentinel lymph node localization in human breast cancer. PMID- 15126608 TI - Skin cancers and non-hodgkin lymphoma among users of systemic glucocorticoids: a population-based cohort study. AB - Patients treated with glucocorticoids may have an increased risk of skin cancer. Using data from the population-based North Jutland Prescription Database and the Danish Cancer Registry, we compared observed and expected numbers of cases of skin cancer and non-Hodgkin lymphoma among 59 043 individuals who received prescriptions for glucocorticoids, a common immunosuppressive therapy, during an 8-year period from January 1, 1989, through December 31, 1996. The overall risks for squamous cell carcinomas and basal cell carcinomas of the skin were increased, particularly among persons who had 15 or more prescriptions (standardized incidence ratio [SIR] for squamous cell carcinomas = 2.45, 95% confidence interval [CI] = 1.37 to 4.04; SIR for basal cell carcinomas = 1.52, 95% CI = 1.09 to 2.07). An elevated risk was also found for non-Hodgkin lymphoma among those with 10-14 prescriptions (SIR = 2.68, 95% CI = 1.16 to 5.29). Our data suggest that use of glucocorticoids may be a shared risk factor for certain skin cancers and lymphomas. PMID- 15126609 TI - Re: Germline BRCA1 mutations and a basal epithelial phenotype in breast cancer. PMID- 15126610 TI - Re: Germline BRCA1 mutations and a basal epithelial phenotype in breast cancer. PMID- 15126613 TI - Re: Effect of epidermal growth factor receptor inhibitor on development of estrogen receptor-negative mammary tumors. PMID- 15126614 TI - Re: Computer-aided detection of breast cancer: has promise outstripped performance? PMID- 15126615 TI - Re: Racial differences in mortality among Medicare recipients after treatment for localized prostate cancer. PMID- 15126618 TI - Polarity establishment in yeast. PMID- 15126619 TI - Molecular mechanisms of E2F-dependent activation and pRB-mediated repression. AB - Alterations in transcription of genes regulated by members of the E2F family of transcription factors can be viewed as a measure of the ebb and flow in a constantly evolving battle between repressor and activator complexes. Various chromatin regulatory complexes have been linked to Rb/E2F proteins, and changes in histone modifications correlate with states of E2F-dependent transcription. E2F has traditionally been viewed in the context of cell-cycle control. However, several recent studies have revealed a new aspect of E2F function in which pRB/E2F-family proteins confer stable repression of transcription. Such repression is evident in both actively proliferating cells and in cells that have withdrawn from the cell cycle. PMID- 15126620 TI - The subapical compartment: a traffic center in membrane polarity development. AB - Spatially separated apical and basolateral plasma membrane domains that have distinct functions and molecular compositions are a characteristic feature of epithelial cell polarity. The subapical compartment (SAC), also known as the common endosome (CE), where endocytic pathways from both surfaces merge, plays a crucial role in the maintenance and probably the biogenesis of these distinct membrane domains. Although differences in morphology are apparent, the same principal features of a SAC can be distinguished in different types of epithelial cells. As polarity develops, the compartment acquires several distinct machineries that, in conjunction with the cytoskeleton, are necessary for polarized trafficking. Disrupting trafficking via the SAC and hence bypassing its sorting machinery, as occurs upon actin depolymerization, leads to mis-sorting of apical and basolateral molecules, thereby compromising the development of polarity. The structural and functional integrity of the compartment in part depends on microtubules. Moreover, the acquisition of a particular set of Rab proteins, including Rab11 and Rab3, appears to be crucial in regulating molecular sorting and vesicular transport relevant both to recycling to either plasma membrane domain and to de novo assembly of the apical domain. Furthermore, subcompartmentalization of the SAC appears to be key to its various functions. PMID- 15126621 TI - Retention and stimulus-dependent recycling of dense core vesicle content in neuroendocrine cells. AB - We have used fluorescence imaging of individual exocytic events in combination with immunogold electron microscopy and FM1-43 photoconversion to study the stimulus-dependent recycling of dense core vesicle content in isolated rat pituitary lactotrophs. Secretory stimulation with high external [K(+)] resulted in 100 exocytic sites per cell that were labeled by extracellular antibodies against the peptide hormone prolactin. Morphological analysis demonstrated that the prolactin was retained and internalized in intact dense cores. Vesicles containing non-secreted, internalized prolactin did not colocalize with DiI-LDL that had been chased into lysosomes but did transiently colocalize with internalized transferrin. The recycling vesicles also trafficked through a syntaxin 6-positive compartment but not the TGN38-positive trans-Golgi. Recycling vesicles, which returned to the cell surface in a slow basal manner, could also be stimulated to undergo exocytosis with a high release probability during subsequent exocytic stimulation with external K(+). These studies suggest a functional role for recycling vesicles that retain prolactin. PMID- 15126622 TI - Calcium transients induce spatially coordinated increases in traction force during the movement of fish keratocytes. AB - The coordination of protrusion with retraction is essential for continuous cell movement. In fish keratocytes the activation of stretch-activated calcium channels, and the resulting increase in intracellular calcium, trigger release of the rear cell margin when forward movement is impeded. Although it is likely that retraction involves a calcium-dependent increase in cytoskeletal contractility, it is not known how the timing, magnitude and localization of contractile forces are organized during retraction. We have addressed this question using a new gelatin traction force assay in combination with calcium imaging to determine what changes in cytoskeletal force production accompany calcium-induced retraction. We find that individual calcium transients are followed within seconds by a rapid increase in traction stress that is maintained, or increases in a stepwise manner, until retraction occurs. Increases in traction stress are accompanied by a distinct sequence of changes in the spatial distribution of large traction stresses. Regions of increased traction stress enlarge at the lateral cell margins and expand forward along the cell margin. In particular, rearward facing propulsive' tractions at the leading edge of the cell, which are normally very low, increase several fold. Following retraction, a precipitous drop in traction stress is observed. Such distinct variations in traction stress are not observed in cells when calcium transients are absent. These results suggest a mechanism by which global increases in intracellular calcium can locally regulate contractile force production, in order to maintain a rapid highly directed mode of movement. PMID- 15126623 TI - Cryptic O2- -generating NADPH oxidase in dendritic cells. AB - All the components of the O(2)(-)-generating NADPH oxidase typically found in neutrophils, namely a membrane-bound low potential flavocytochrome b and oxidase activation factors of cytosolic origin, are immunodetectable in murine dendritic cells (DCs). However, in contrast to neutrophils, DCs challenged with phorbol myristate acetate (PMA) can barely mount a significant respiratory burst. Nevertheless, DCs generate a substantial amount of O(2)(-) in the presence of PMA following preincubation with pro-inflammatory ligands such as lipopolysaccharide and pansorbin, and to a lesser extent with anti-CD40 or polyinosinic polycytidylic acid. We found that the virtual lack of the oxidase response to PMA alone is specifically controlled in DCs. Through the use of homologous and heterologous cell-free systems of oxidase activation, we showed the following: (1) a NADPH oxidase inhibitory factor is located in DC membranes; it exerts its effect on oxidase activation and not on the activated oxidase. (2) The inhibition is relieved by pretreatment of DC membranes with beta-octylglucoside (beta-OG). (3) The beta-OG-extracted inhibitory factor prevents the activation of neutrophil oxidase. (4) The inhibitory activity is lost after treatment of DC membranes with proteinase K or heating, which points to the protein nature of the inhibitory factor. Overall, these data indicate that the O(2)(-)-generating oxidase in DCs is cryptic, owing to the presence of a membrane-bound inhibitor of protein nature that prevents oxidase activation. The inhibition is relieved under specific conditions, including a prolonged contact of DCs with pro-inflammatory ligands from microbial origin, allowing a substantial production of O(2)(-), which may contribute to the response of DCs to a microbial exposure. PMID- 15126624 TI - Overexpression of the C-terminal PG-M/versican domain impairs growth of tumor cells by intervening in the interaction between epidermal growth factor receptor and beta1-integrin. AB - Versican is highly expressed in many types of tumors. In a previous study, we found that a G3 mutant [G3DeltaEGF; a versican G3 domain lacking two epidermal growth factor (EGF)-like motifs] exerted a dominant-negative effect on versican secretion and binding. Here, we report that astrocytoma U87 cells expressing the versican G3 mutant lost the hallmark of cell transformation and tumorigenesis in vitro and in vivo. U87 cells expressing G3DeltaEGF had enhanced cell adhesion and spreading, but lost the tumor characteristic of anchorage-independent growth. When U87 cells were deprived of serum, FAK was quickly dephosphorylated, integrin/EGF-receptor (EGFR) complexes dissociated and the cells retained an appropriate level of EGFR phosphorylation. These cells quickly detached, migrated, rounded, reorganized and survived. However, after serum withdrawal from G3DeltaEGF-transfected U87 cells, sustained FAK phosphorylation and integrin-EGFR association were observed, but a greatly reduced EGFR phosphorylation. These cells remained spread and continued to grow before undergoing massive apoptosis. The addition of EGF promoted U87 cell rounding but had little effect on G3DeltaEGF-transfected cells owing to reduced EGFR phosphorylation. Our study sheds light on the question of how the matrix molecule versican modulates tumorigenesis by affecting integrin and EGFR signals. PMID- 15126625 TI - Complexin I regulates glucose-induced secretion in pancreatic beta-cells. AB - The neuronal-specific protein complexin I (CPX I) plays an important role in controlling the Ca(2+)-dependent neurotransmitter release. Since insulin exocytosis and neurotransmitter release rely on similar molecular mechanisms and that pancreatic beta-cells and neuronal cells share the expression of many restricted genes, we investigated the potential role of CPX I in insulin secreting cells. We found that pancreatic islets and several insulin-secreting cell lines express high levels of CPX I. The beta-cell expression of CPX I is mediated by the presence of a neuron restrictive silencer element located within the regulatory region of the gene. This element bound the transcriptional repressor REST, which is found in most cell types with the exception of mature neuronal cells and beta-cells. Overexpression of CPX I or silencing of the CPX I gene (Cplx1) by RNA interference led to strong impairment in beta-cell secretion in response to nutrients such as glucose, leucine and KCl. This effect was detected both in the early and the sustained secretory phases but was much more pronounced in the early phase. We conclude that CPX I plays a critical role in beta-cells in the control of the stimulated-exocytosis of insulin. PMID- 15126626 TI - Processing and activation of latent heparanase occurs in lysosomes. AB - Heparanase is a heparan sulfate degrading endoglycosidase participating in extracellular matrix degradation and remodeling. Heparanase is synthesized as a 65 kDa non-active precursor that subsequently undergoes proteolytic cleavage, yielding 8 kDa and 50 kDa protein subunits that heterodimerize to form an active enzyme. The protease responsible for heparanase processing is currently unknown, as is the sub-cellular processing site. In this study, we characterize an antibody (733) that preferentially recognizes the active 50 kDa heparanase form as compared to the non-active 65 kDa heparanase precursor. We have utilized this and other anti-heparanase antibodies to study the cellular localization of the latent 65 kDa and active 50 kDa heparanase forms during uptake and processing of exogenously added heparanase. Interestingly, not only the processed 50 kDa, but also the 65 kDa heparanase precursor was localized to perinuclear vesicles, suggesting that heparanase processing occurs in lysosomes. Indeed, heparanase processing was completely inhibited by chloroquine and bafilomycin A1, inhibitors of lysosome proteases. Similarly, processing of membrane-targeted heparanase was also chloroquine-sensitive, further ruling out the plasma membrane as the heparanase processing site. Finally, we provide evidence that antibody 733 partially neutralizes the enzymatic activity of heparanase, suggesting that the N terminal region of the molecule is involved in assuming an active conformation. Monoclonal antibodies directed to this region are likely to provide specific heparanase inhibitors and hence assist in resolving heparanase functions under normal and pathological conditions. PMID- 15126628 TI - A novel isoform of sarcolemmal membrane-associated protein (SLMAP) is a component of the microtubule organizing centre. AB - The microtubule organizing centre (MTOC) or the centrosome serves a crucial role in the establishment of cellular polarity, organization of interphase microtubules and the formation of the bipolar mitotic spindle. We have elucidated the genomic structure of a gene encoding the sarcolemmal membrane-associated protein (SLMAP), which encodes a 91 kDa polypeptide with a previously uncharacterized N-terminal sequence encompassing a forkhead-associated (FHA) domain that resides at the centrosome. Anti-peptide antibodies directed against SLMAP N-terminal sequences showed colocalization with gamma-tubulin at the centrosomes at all phases of the cell cycle. Agents that specifically disrupt microtubules did not affect SLMAP association with centrosomes. Furthermore, SLMAP sequences directed a reporter green fluorescent protein (GFP) to the centrosome, and deletions of the newly identified N-terminal sequence from SLMAP prevented the centrosomal targeting. Deletion-mutant analysis concluded that overall, structural determinants in SLMAP were responsible for centrosomal targeting. Elevated levels of centrosomal SLMAP were found to be lethal, whereas mutants that lacked centrosomal targeting inhibited cell growth accompanied by an accumulation of cells at the G2/M phase of the cell cycle. PMID- 15126627 TI - CRM1-dependent, but not ARE-mediated, nuclear export of IFN-alpha1 mRNA. AB - While the bulk of cellular mRNA is known to be exported by the TAP pathway, export of specific subsets of cellular mRNAs may rely on chromosome region maintenance 1 (CRM1). One line of evidence supporting this hypothesis comes from the study of mRNAs of certain early response genes (ERGs) containing the adenylate uridylate-rich element (ARE) in their 3' untranslated regions (3' UTRs). It was reported that HuR-mediated nuclear export of these mRNAs was CRM1 dependent under certain stress conditions. To further examine potential CRM1 pathways for other cellular mRNAs under stress conditions, the nuclear export of human interferon-alpha1 (IFN-alpha1) mRNA, an ERG mRNA induced upon viral infection, was studied. Overproduction of human immunodeficiency virus type 1 Rev protein reduced the expression level of the co-transfected IFN-alpha1 gene. This inhibitory effect, resulting from nuclear retention of IFN-alpha1 mRNA, was reversed when rev had a point mutation that made its nuclear export signal unable to associate with CRM1. Leptomycin B sensitivity experiments revealed that the cytoplasmic expression of IFN-alpha1 mRNA was arrested upon inhibition of CRM1. This finding was further supported by overexpression of DeltaCAN, a defective form of the nucleoporin Nup214/CAN that inhibits CRM1 in a dominant-negative manner, which resulted in the effective inhibition of IFN-alpha1 gene expression. Subsequent RNA fluorescence in situ hybridisation and immunocytochemistry demonstrated that the IFN-alpha1 mRNA was colocalised with CRM1, but not with TAP, in the nucleus. These results therefore imply that the nuclear export of IFN alpha1 mRNA is mediated by CRM1. However, truncation of the 3' UTR did not negatively affect the nuclear export of IFN-alpha1 mRNA that lacked the ARE, unexpectedly indicating that this CRM1-dependent mRNA export may not be mediated via the ARE. PMID- 15126629 TI - Suppression of a mitotic mutant by tRNA-Ala anticodon mutations that produce a dominant defect in late mitosis. AB - Cold-sensitive dominant mutants scn1 and scn2 of Schizosaccharomyces pombe were isolated by their ability to suppress temperature-sensitive cut9-665 defective in an essential subunit (human Apc6/budding yeast Cdc16 ortholog) of anaphase promoting complex/cyclosome (APC/C). APC/C mutants were defective in metaphase/anaphase transition, whereas single scn mutants showed the delay in anaphase spindle elongation at 20 degrees C. The scn mutants lost viability because of chromosome missegregation, and were sensitive to a tubulin poison. To understand the scn phenotypes, mutant genes were identified. Surprisingly, scn1 and scn2 have the same substitution in the anticodon of two different tRNA-Ala (UGC) genes. UGC was altered to UGU so that the binding of the tRNA-Ala to the ACA Thr codon in mRNA became possible. As cut9-665 contained an Ala535Thr substitution, wild-type Cut9 protein was probably produced in scn mutants. Indeed, plasmid carrying tRNA-Ala (UGU) conferred cold-sensitivity to wild-type and suppressed cut9-665 in a dominant fashion. The previously identified scn1(+) (renamed as scn3(+)) turned out to be a high copy suppressor for scn1 and scn2. These are the first tRNA mutants that cause a mitotic defect. PMID- 15126630 TI - Cathepsin D is involved in the regulation of transglutaminase 1 and epidermal differentiation. AB - We previously demonstrated that the aspartate protease cathepsin D is activated by ceramide derived from acid sphingomyelinase. Increased expression of cathepsin D in the skin has been reported in wound healing, psoriasis and skin tumors. We explored specific functions of cathepsin D during epidermal differentiation. Protein expression and enzymatic activity of cathepsin D increased in differentiated keratinocytes in both stratified organotypic cultures and in mouse skin during epidermal barrier repair. Treatment of cultured keratinocytes with exogenous cathepsin D increased the activity of transglutaminase 1, known to cross-link the cornified envelope proteins involucrin and loricrin during epidermal differentiation. Inhibition of cathepsin D by pepstatin A suppressed the activity of transglutaminase 1. Cathepsin D-deficient mice revealed reduced transglutaminase 1 activity and reduced protein levels of the cornified envelope proteins involucrin and loricrin. Also, amount and distribution of cornified envelope proteins involucrin, loricrin, filaggrin, and of the keratins K1 and K5 were significantly altered in cathepsin D-deficient mice. Stratum corneum morphology in cathepsin D-deficient mice was impaired, with increased numbers of corneocyte layers and faint staining of the cornified envelope only, which is similar to the human skin disease lamellar ichthyosis. Our findings suggest a functional link between cathepsin D activation, transglutaminase 1 activity and protein expression of cornified envelope proteins during epidermal differentiation. PMID- 15126631 TI - Yeast ARL1 encodes a regulator of K+ influx. AB - A molecular genetic approach was undertaken in Saccharomyces cerevisiae to examine the functions of ARL1, encoding a G protein of the Ras superfamily. We show here that ARL1 is an important component of the control of intracellular K(+). The arl1 mutant was sensitive to toxic cations, including hygromycin B and other aminoglycoside antibiotics, tetramethylammonium ions, methylammonium ions and protons. The hygromycin-B-sensitive phenotype was suppressed by the inclusion of K(+) and complemented by wild-type ARL1 and an allele of ARL1 predicted to be unbound to nucleotide in vivo. The arl1 mutant strain internalized approximately 25% more [(14)C]-methylammonium ion than did the wild type, consistent with hyperpolarization of the plasma membrane. The arl1 strain took up 30-40% less (86)Rb(+) than did the wild type, showing an inability to regulate K(+) import properly, contributing to membrane hyperpolarity. By contrast, K(+) and H(+) efflux were undisturbed. The loss of ARL1 had no effect on the steady-state level or the localization of a tagged version of Trk1p. High copy suppressors of the hygromycin-B phenotype included SAP155, encoding a protein that interacts with the cell cycle regulator Sit4p, and HAL4 and HAL5, encoding Ser/Thr kinases that regulate the K(+)-influx mediators Trk1p and Trk2p. These results are consistent with a model in which ARL1, via regulation of HAL4/HAL5, governs K(+) homeostasis in cells. PMID- 15126632 TI - Efficient endosome-to-Golgi transport of Shiga toxin is dependent on dynamin and clathrin. AB - It has previously been shown that Shiga toxin, despite being bound to a glycolipid receptor, can be efficiently endocytosed from clathrin-coated pits. However, clathrin-independent endocytosis is also responsible for a proportion of the toxin uptake in some cells. After endocytosis the toxin can be transported in retrograde fashion to the Golgi apparatus and the endoplasmic reticulum, and then to the cytosol, where it exerts its toxic effect by inactivating ribosomes. In order to investigate the role of dynamin and clathrin in endosome-to-Golgi transport of Shiga toxin, we have used HeLa dyn(K44A) and BHK antisense clathrin heavy chain (CHC) cells that, in an inducible manner, express mutant dynamin or CHC antisense RNA, respectively. In these cell lines, one can study the role of dynamin and clathrin on endosome-to-Golgi transport because they, as shown here, still internalize Shiga toxin when dynamin- and clathrin-dependent endocytosis is blocked. Butyric acid has been shown to sensitize A431 cells to Shiga toxin by increasing the proportion of cell-associated toxin that is transported to the Golgi apparatus and the endoplasmic reticulum. Here, we find that, in HeLa and BHK cells also, butyric acid also increased toxin transport to the Golgi apparatus and sensitized the cells to Shiga toxin. We have therefore studied the role of dynamin and clathrin in both untreated and butyric-acid-treated cells by measuring the sulfation of a modified Shiga B fragment. Our results indicate that endosome-to-Golgi transport of Shiga toxin is dependent on functional dynamin in both untreated cells and in cells treated with butyric acid. Interestingly, the regulation of Shiga toxin transport in untreated and butyric-acid-treated cells differs when it comes to the role of clathrin, because only cells that are sensitized to Shiga toxin with butyric acid need functional clathrin for endosome to-Golgi transport. PMID- 15126633 TI - Cross-linking ATP synthase complexes in vivo eliminates mitochondrial cristae. AB - We have used the tetrameric nature of the fluorescent protein DsRed to cross-link F(1)F(O)-ATPase complexes incorporating a subunit gamma-DsRed fusion protein in vivo. Cells expressing such a fusion protein have impaired growth relative to control cells. Strikingly, fluorescence microscopy of these cells revealed aberrant mitochondrial morphology. Electron microscopy of cell sections revealed the absence of cristae and multiple layers of unfolded inner mitochondrial membrane. Complexes recovered from detergent lysates of mitochondria were present largely as tetramers. Co-expression of 'free' DsRed targeted to the mitochondria reduced F(1)F(O)-ATPase oligomerisation and partially reversed the impaired growth and abnormal mitochondrial morphology. We conclude that the correct arrangement of F(1)F(O)-ATPase complexes within the mitochondrial inner membrane is crucial for the genesis and/or maintenance of mitochondrial cristae and morphology. Our findings further suggest that F(1)F(O)-ATPase can exist in oligomeric associations within the membrane during respiratory growth. PMID- 15126634 TI - Membrane fusion of secretory vesicles of the sea urchin egg in the absence of NSF. AB - The role of cytosolic ATPases such as N-ethylmaleimide (NEM)-sensitive fusion protein (NSF) in membrane fusion is controversial. We examined the physiology and biochemistry of ATP and NSF in the cortical system of the echinoderm egg to determine if NSF is an essential factor in membrane fusion during Ca(2+) triggered exocytosis. Neither exocytosis in vitro, nor homotypic cortical vesicle (CV) fusion required soluble proteins or nucleotides, and both occurred in the presence of non-hydrolyzable analogs of ATP. While sensitive to thiol-specific reagents, CV exocytosis is not restored by the addition of cytosolic NSF, and fusion and NSF function are differentially sensitive to thiol-specific agents. To test participation of tightly bound, non-exchangeable NSF in CV-CV fusion, we cloned the sea urchin homolog and developed a species-specific antibody for western blots and physiological analysis. This antibody was without effect on CV exocytosis or homotypic fusion, despite being functionally inhibitory. NSF is detectable in intact cortices, cortices from which CVs had been removed and isolated CVs treated with ATP-gamma-S and egg cytosol to reveal NSF binding sites. In contrast, isolated CVs, though all capable of Ca(2+)-triggered homotypic fusion, contain less than one hexamer of NSF per CV. Thus NSF is not a required component of the CV fusion machinery. PMID- 15126635 TI - Nedd4.1-mediated ubiquitination and subsequent recruitment of Tsg101 ensure HTLV 1 Gag trafficking towards the multivesicular body pathway prior to virus budding. AB - One of the most exciting recent developments in the field of retroviruses is the finding that their Gag proteins hijack cellular proteins from the mutivesicular body (MVB) pathway during the budding process. The Gag proteins of oncoretroviruses possess a PPxY motif that recruits a ubiquitin ligase from the Nedd4 family, whereas those of the human immunodeficiency virus interact through a PTAP motif with Tsg101, a protein of the ESCRT-1 complex. It is currently assumed that Nedd4 and Tsg101 represent equivalent entry gates towards the same cellular process leading to budding, and that both partners are recruited to the plasma membrane where viral budding occurs. However, we report here that the budding of the human oncoretrovirus HTLV-1, the Gag proteins of which possess tandem PPPY/PTAP motifs, requires both Nedd4 and Tsg101. We show that Nedd4.1, but not Nedd4.2, is recruited by the PPPY motif of Gag and subsequently catalyzes Gag ubiquitination. We also demonstrate that Gag interacts first with Nedd4.1 at the plasma membrane and then with Tsg101 in late endosomes/MVBs. Consistently, we found that HTLV-1 particles mutated in the PPPY motif remain underneath the plasma membrane, blocked at an early step of the budding process, whereas PTAP mutated viruses accumulate in intracellular vesicles, blocked at a later step. Our findings indicate that Nedd4.1 and Tsg101 act successively in the assembly process of HTLV-1 to ensure proper Gag trafficking through the endocytic pathway up to late endosomes where the late steps of retroviral release occur. PMID- 15126636 TI - Expression of the endocytic proteins dynamin and amphiphysin in rat gastric enterochromaffin-like cells. AB - Dynamin and amphiphysin play crucial roles in a variety of endocytic processes. Previous investigations of expression and functions of these proteins were performed mostly on neurons. The aim of this study was to investigate the presence and interaction of dyn and amph in gastric enterochromaffin-like cells. These endocrine cells of the gastric mucosa play a pivotal role in the regulation of acid secretion. Exocytosis of histamine-containing secretory vesicles has been described in detail. However, the mechanisms of endocytosis are unknown in this neuroendocrine cell type. Using RT-PCR and western blotting, we detected dynamin 1, -2 and -3 in highly enriched isolated enterochromaffin-like cells. Dynamin-1 and -2 were expressed at similar high levels, whereas dynamin-3 was of low abundance. Immunofluorescence microscopy located dynamin-1 and -2 to the cytoplasm and cell surface, whereas dynamin-3 was distributed differently in the perinuclear area. The presence of amphiphysin-1 and -2 RNAs was revealed by RT PCR and a new splice variant of amphiphysin-2 was detected. Amphiphysin-1 and -2 were also detected in enterochromaffin-like cells by immunohistochemistry in the same locations as dynamin-1 and -2. Amphiphysin-1 and dynamin-1 co immunoprecipitated with amphiphysin-2. In addition, dynamin-1 and amphiphysin-2 partially colocalized at the plasma membrane. Our results confirm the interaction of dynamin and amphiphysin and imply a role in endocytosis in enterochromaffin like cells. To our knowledge, this is the first demonstration of the co expression of all three dynamin isoforms in a non-tumor cell. PMID- 15126637 TI - PDE4-regulated cAMP degradation controls the assembly of integrin-dependent actin adhesion structures and REF52 cell migration. AB - Plating of REF52 cells onto extracellular matrix components leads to the formation of integrin-dependent actin adhesion microspikes. We show that the formation of these structures is sensitive to chemical agents that regulate cAMP levels, such as forskolin and IBMX. In particular, by using the specific inhibitor rolipram, we identify the PDE4 family of cAMP-specific phosphodiesterases as critical regulators of this process. The effect of PDE4 on microspike formation is mediated by actions exerted through the activation of PKA - rather than through the alternative cAMP effector, Epac. We provide evidence that peripheral microspikes are RhoA-, ROCK- and myosin-dependent, and that this pathway is suppressed by PDE4 inhibition. In addition, PDE4 inhibition impairs cell locomotion that requires dynamic protrusion and retraction of peripheral spike structures. Our data demonstrate that PDE4 activity is a key modulator of integrin-induced actin assembly at the cell periphery which, in turn, controls cell migration. PMID- 15126638 TI - The GDP-bound form of Arf6 is located at the plasma membrane. AB - The function of Arf6 has been investigated largely by using the T27N and the Q67L mutants, which are thought to be blocked in GDP- and GTP-bound states, respectively. However, these mutants have been poorly characterized biochemically. Here, we found that Arf6(T27N) is not an appropriate marker of the inactive GDP-bound form because it has a high tendency to lose its nucleotide in vitro and to denature. As a consequence, most of the protein is aggregated in vivo and localizes to detergent-insoluble structures. However, a small proportion of Arf6(T27N) is able to form a stable complex with its exchange factor EFA6 at the plasma membrane, accounting for its dominant-negative phenotype. To define the cellular localization of Arf6-GDP, we designed a new mutant, Arf6(T44N). In vitro, this mutant has a 30-fold decreased affinity for GTP. In vivo, it is mostly GDP bound and, in contrast to the wild type, does not switch to the active conformation when expressed with EFA6. This GDP-locked mutant is found at the plasma membrane, where it localizes with EFA6 and Ezrin in actin- and phosphatidylinositol (4,5)-bisphosphate-enriched domains. From these results, we conclude that the Arf6 GDP-GTP cycle takes place at the plasma membrane. PMID- 15126639 TI - Chloroplast division site placement requires dimerization of the ARC11/AtMinD1 protein in Arabidopsis. AB - Chloroplast division is mediated by the coordinated action of a prokaryote derived division system(s) and a host eukaryote-derived membrane fission system(s). The evolutionary conserved prokaryote-derived system comprises several nucleus-encoded proteins, two of which are thought to control division site placement at the midpoint of the organelle: a stromal ATPase MinD and a topological specificity factor MinE. Here, we show that arc11, one of 12 recessive accumulation and replication of chloroplasts (arc) mutants in Arabidopsis, contains highly elongated and multiple-arrayed chloroplasts in developing green tissues. Genomic sequence analysis revealed that arc11 contains a missense mutation in alpha-helix 11 of the chloroplast-targeted AtMinD1 changing an Ala at position 296 to Gly (A296G). Introduction of wild-type AtMinD1 restores the chloroplast division defects of arc11 and quantitative RT-PCR analysis showed that the degree of complementation was highly dependent on transgene expression levels. Overexpression of the mutant ARC11/AtMinD1 in transgenic plants results in the inhibition of chloroplast division, showing that the mutant protein has retained its division inhibition activity. However, in contrast to the defined and punctate intraplastidic localization patterns of an AtMinD1-YFP fusion protein, the single A296G point mutation in ARC11/AtMinD1 results in aberrant localization patterns inside chloroplasts. We further show that AtMinD1 is capable of forming homodimers and that this dimerization capacity is abolished by the A296G mutation in ARC11/AtMinD1. Our data show that arc11 is a loss-of-function mutant of AtMinD1 and suggest that the formation of functional AtMinD1 homodimers is paramount for appropriate AtMinD1 localization, ultimately ensuring correct division machinery placement and chloroplast division in plants. PMID- 15126640 TI - Scrapie-like prion protein is translocated to the nuclei of infected cells independently of proteasome inhibition and interacts with chromatin. AB - Prion diseases are fatal transmissible neurodegenerative disorders characterized by the accumulation of an abnormally folded isoform of the cellular prion protein (PrP(C)) denoted PrP(Sc). Recently, wild-type and pathogenic PrP mutants have been shown to be degraded by the endoplasmic reticulum-associated degradation proteasome pathway after translocation into the cytosol. We show here that a protease resistant form of PrP accumulated in the nuclei of prion-infected cells independently of proteasome activity, and that this nuclear translocation required an intact microtubule network. Moreover, our results show for the first time that nuclear PrP interacts with chromatin in vivo, which may have physiopathological consequences in prion diseases PMID- 15126641 TI - Chronic oxidative stress compromises telomere integrity and accelerates the onset of senescence in human endothelial cells. AB - Replicative senescence and oxidative stress have been implicated in ageing, endothelial dysfunction and atherosclerosis. Replicative senescence is determined primarily by telomere integrity. In endothelial cells the glutathione redox-cycle plays a predominant role in the detoxification of peroxides. The aim of this study was to elucidate the role of the glutathione-dependent antioxidant system on the replicative capacity and telomere dynamics of cultured endothelial cells. Human umbilical vein endothelial cells were serially passaged while exposed to regular treatment with 0.1 microM tert-butyl hydroperoxide, a substrate of glutathione peroxidase, or 10 microM L-buthionine-[S,R]-sulphoximine, an inhibitor of glutathione synthesis. Both treatments induced intracellular oxidative stress but had no cytotoxic or cytostatic effects. Nonetheless, treated cultures entered senescence prematurely (30 versus 46 population doublings), as determined by senescence-associated beta-galactosidase staining and a sharp decrease in cell density at confluence. In cultures subjected to oxidative stress terminal restriction fragment (TRF) analysis demonstrated faster telomere shortening (110 versus 55 bp/population doubling) and the appearance of distinct, long TRFs after more than 15-20 population doublings. Fluorescence in situ hybridisation analysis of metaphase spreads confirmed the presence of increased telomere length heterogeneity, and ruled out telomeric end-to-end fusions as the source of the long TRFs. The latter was also confirmed by Bal31 digestion of genomic DNA. Similarly, upregulation of telomerase could not account for the appearance of long TRFs, as oxidative stress induced a rapid and sustained decrease in this activity. These findings demonstrate a key role for glutathione dependent redox homeostasis in the preservation of telomere function in endothelial cells and suggest that loss of telomere integrity is a major trigger for the onset of premature senescence under mild chronic oxidative stress. PMID- 15126642 TI - Altered pharmacology of synaptic and extrasynaptic GABAA receptors on CA1 hippocampal neurons is consistent with subunit changes in a model of alcohol withdrawal and dependence. AB - Previously, we reported (Cagetti, Liang, Spigelman, and Olsen, 2003) that chronic intermittent ethanol (CIE) treatment leads to signs of alcohol dependence, including anxiety and hyperactivity, accompanied by reduced synaptic gamma aminobutyric acid (A) receptor (GABAAR) function and altered sensitivity to its allosteric modulators consistent with a measured switch in subunit composition. In this study, we separated the synaptic and extrasynaptic components of GABAAR activation in recordings from pyramidal CA1 cells of hippocampal slices and demonstrated marked differences in the responsiveness of synaptic and extrasynaptic GABAARs to agonists and allosteric modulators in control rats, and in the way they are altered following CIE treatment. Notably, tonic inhibition mediated by extrasynaptic GABAARs was differentially sensitive to the partial agonist gaboxadol (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol; THIP) and the allosteric modulator zolpidem, compared with the miniature inhibitory synaptic currents (mIPSCs) in the same cells from saline-treated rats. After CIE treatment, potentiation of tonic currents by diazepam and zolpidem was lost, whereas potentiation by the alpha4 subunit-preferring benzodiazepine Ro15-4513 (ethyl 8-azido-6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a]-[1,4]benzodiazepine-3 carboxylate) and THIP was only partially reduced. Potentiation of synaptic GABAAR currents by zolpidem was eliminated after CIE, whereas THIP slightly inhibited mIPSCs from control rats and greatly enhanced them after CIE treatment. These results are consistent with alpha1 subunit decreases at synaptic and extrasynaptic GABAARs, whereas alpha4 subunits are increased at synaptic and decreased at extrasynaptic GABAARs. Behaviorally, THIP was active as a hypnotic and anxiolytic but not as an anti-convulsant against pentylenetetrazol seizures in control rats. Only slight tolerance was observed to the sleep time, but not to the anxiolytic, effect of THIP after CIE. Thus, differential alterations in synaptic and extrasynaptic GABAARs appear to play an important role in the brain plasticity of alcohol dependence, and withdrawal signs may be profitably treated with GABAergic drugs such as THIP, which does not show cross-tolerance with ethanol. PMID- 15126644 TI - Cardiac transcriptional response to acute and chronic angiotensin II treatments. AB - Exposure of experimental animals to increased angiotensin II (ANG II) induces hypertension associated with cardiac hypertrophy, inflammation, and myocardial necrosis and fibrosis. Some of the most effective antihypertensive treatments are those that antagonize ANG II. We investigated cardiac gene expression in response to acute (24 h) and chronic (14 day) infusion of ANG II in mice; 24-h treatment induces hypertension, and 14-day treatment induces hypertension and extensive cardiac hypertrophy and necrosis. For genes differentially expressed in response to ANG II treatment, we tested for significant regulation of pathways, based on Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Microarray Pathway Profiler (GenMAPP) databases, as well as functional classes based on Gene Ontology (GO) terms. Both acute and chronic ANG II treatments resulted in decreased expression of mitochondrial metabolic genes, notably those for the electron transport chain and Krebs-TCA cycle; chronic ANG II treatment also resulted in decreased expression of genes involved in fatty acid metabolism. In contrast, genes involved in protein translation and ribosomal activity increased expression following both acute and chronic ANG II treatments. Some classes of genes showed differential response between acute and chronic ANG II treatments. Acute treatment increased expression of genes involved in oxidative stress and amino acid metabolism, whereas chronic treatments increased cytoskeletal and extracellular matrix genes, second messenger cascades responsive to ANG II, and amyloidosis genes. Although a functional linkage between Alzheimer disease, hypertension, and high cholesterol has been previously documented in studies of brain tissue, this is the first demonstration of induction of Alzheimer disease pathways by hypertension in heart tissue. This study provides the most comprehensive available survey of gene expression changes in response to acute and chronic ANG II treatment, verifying results from disparate studies, and suggests mechanisms that provide novel insight into the etiology of hypertensive heart disease and possible therapeutic interventions that may help to mitigate its effects. PMID- 15126643 TI - Regional hemodynamic effects of the N-(2-benzoylphenyl)-L-tyrosine peroxisome proliferator-activated receptor-gamma ligand, GI 262570 [(S)-2-(2 benzoylphenylamino)-3-[4-[2-(5-methyl-2-phenyl-2-oxazol-4 yl)ethoxy]phenyl]propionic acid], in conscious rats. AB - This study provides novel data on the regional hemodynamic effects of the peroxisome proliferator-activated receptor-gamma activator, GI 262570 [(S)-2-(2 benzoylphenylamino)-3-[4-[2-(5-methyl-2-phenyl-2-oxazol-4 yl)ethoxy]phenyl]propionic acid], in conscious, male Sprague-Dawley rats. Administration of GI 262570 twice daily for 4 days caused a slowly developing, modest fall in mean arterial blood pressure, associated with a progressive, hyperemic hindquarters vasodilatation, but with no consistent changes in renal or mesenteric hemodynamics. The hindquarters vasodilator effect of GI 262570 was not inhibited by the beta2-adrenoceptor antagonist, ICI 118551 ((+/-)-1-[2,3-(dihydro 7-methyl-1H-inden-4-yl)oxy]-3-[(1-methylethyl) amino]-2-butanol hydrochloride), and was still apparent in the presence of the alpha-adrenoceptor antagonist, phentolamine. Neither the latter, nor antagonism of angiotensin (AT1) and endothelin (ETA and ETB) receptors unmasked vasodilator responses to GI 262570 in the renal or mesenteric vascular beds. In the presence of GI 262570, vasodilator responses to acetylcholine and vasoconstrictor responses to methoxamine were normal. Furthermore, the cardiovascular responses to nonselective nitric-oxide synthase inhibition were not influenced by GI 262570. Collectively, these results indicate that the vasodilator action of GI 262570 is specific to the hindquarters vascular bed (of those studied), does not involve alpha- or beta2-adrenoceptors, and is not associated with a change in basal or stimulated nitric oxide release. PMID- 15126645 TI - Neural system-enriched gene expression: relationship to biological pathways and neurological diseases. AB - To understand the commitment of the genome to nervous system differentiation and function, we sought to compare nervous system gene expression to that of a wide variety of other tissues by gene expression database construction and mining. Gene expression profiles of 10 different adult nervous tissues were compared with that of 72 other tissues. Using ANOVA, we identified 1,361 genes whose expression was higher in the nervous system than other organs and, separately, 600 genes whose expression was at least threefold higher in one or more regions of the nervous system compared with their median expression across all organs. Of the 600 genes, 381 overlapped with the 1,361-gene list. Limited in situ gene expression analysis confirmed that identified genes did represent nervous system enriched gene expression, and we therefore sought to evaluate the validity and significance of these top-ranked nervous system genes using known gene literature and gene ontology categorization criteria. Diverse functional categories were present in the 381 genes, including genes involved in intracellular signaling, cytoskeleton structure and function, enzymes, RNA metabolism and transcription, membrane proteins, as well as cell differentiation, death, proliferation, and division. We searched existing public sites and identified 110 known genes related to mental retardation, neurological disease, and neurodegeneration. Twenty-one of the 381 genes were within the 110-gene list, compared with a random expectation of 5. This suggests that the 381 genes provide a candidate set for further analyses in neurological and psychiatric disease studies and that as a field, we are as yet, far from a large-scale understanding of the genes that are critical for nervous system structure and function. Together, our data indicate the power of profiling an individual biologic system in a multisystem context to gain insight into the genomic basis of its structure and function. PMID- 15126646 TI - Managing mineral balance in end-stage renal disease. PMID- 15126647 TI - Importance of hyperphosphataemia in the cardio-renal axis. AB - Hyperphosphataemia occurs in nearly all patients with end-stage renal disease (ESRD). In the past, the need to manage hyperphosphataemia focused primarily on its role as a contributor to secondary hyperparathyroidism and renal osteodystrophy. There is now widespread recognition that disturbances in phosphorus metabolism and/or the therapeutic measures used to manage it are important risk factors for cardiovascular calcification. This serious complication of chronic kidney disease may contribute to the very high mortality rate from cardiovascular causes in patients undergoing long-term dialysis. New strategies for controlling serum phosphorus levels and for better management of mineral metabolism in general are required to address these issues in patients with ESRD. PMID- 15126648 TI - The importance of bone health in end-stage renal disease: out of the frying pan, into the fire? AB - In the early stages of renal failure, hyperparathyroidism develops as a compensatory mechanism to control serum levels of calcium, phosphorus and calcitriol. As kidney disease progresses, this ability to maintain mineral homeostasis is lost, leading to the development of renal osteodystrophy (ROD). Over the past decade, the pattern of ROD seen in patients with chronic kidney disease (CKD) has changed. Previously, the majority of patients had mixed uraemic osteodystrophy or aluminium-related osteomalacia. The decreased use of aluminium based phosphate binders, coupled with improvements in the management of hyperphosphataemia, led to a reduction in the prevalence of these types of ROD. Since the mid-1990s, there has been an increase in the prevalence of adynamic bone disease as a result of increased suppression of parathyroid hormone through the use of calcium-based phosphate binders and calcitriol therapy. Adynamic bone disease is also associated with several clinical factors, such as older age, use of continuous ambulatory peritoneal dialysis and the presence of diabetes mellitus, as well as the use of calcitriol therapy. Studies of calcium metabolism in patients with CKD have shown that adynamic bone disease is a distinct clinical condition that leads to hypercalcaemia via mechanisms different from that seen in high-turnover bone disease. As high calcium x phosphorus product has been associated with soft tissue and vascular calcifications, and increased mortality, optimizing bone health may be an important way of reducing cardiovascular risk in patients with CKD. To do this, novel, effective, non-calcium, non-aluminium phosphate binders will be necessary. PMID- 15126649 TI - Improving outcomes in hyperphosphataemia. AB - Preclinical studies have shown that lanthanum has a very high phosphate-binding capacity at gastrointestinal pH, while clinical trials have shown lanthanum carbonate to be an effective, well-tolerated phosphate binder for the treatment of hyperphosphataemia in patients with end-stage renal disease. Optimization of bone health is an important issue in these patients, and, based on theoretical grounds, there have been concerns that lanthanum will have toxic effects on bone similar to those of aluminium. However, compared with aluminium, absorption of lanthanum is extremely low and lanthanum treatment is not associated with systemic toxicity. In addition, unlike aluminium, elimination of lanthanum is not through the kidney, but mainly takes place via the biliary route and is, therefore, independent of renal function. This implies that patients with chronic renal failure are not at an increased risk for accumulation of the element, compared with patients with normal renal function. In animal studies, no adverse effects on bone were seen in healthy animals receiving lanthanum carbonate. In 5/6th nephrectomized rats, very high doses of lanthanum (1000-2000 mg/kg) affected bone mineralization. This was not due to a direct toxic effect on bone, but was secondary to phosphate depletion induced by lanthanum and, as with any gastro-intestinal phosphate-binding agent, can be reversed with a phosphate supplemented diet. In a phase III clinical trial, bone biopsies were taken from dialysis patients at baseline and after 1 year of treatment with either lanthanum carbonate (median dose, 1250 mg/day) or calcium carbonate (median dose, 2000 mg/day). Patients treated with lanthanum carbonate for 1 year did not experience any of the aluminium-like toxic effects on bone expressed as either osteomalacia or adynamic bone disease. PMID- 15126650 TI - Improving phosphate-binder therapy as a way forward. AB - Aluminium- or calcium-based phosphate-binding agents traditionally have been used to treat hyperphosphataemia in patients with end-stage renal disease. Although these agents effectively lower serum phosphorus levels, they are associated with serious side effects. Aluminium-based agents are associated with bone toxicity, renal osteodystrophy and encephalopathy, and calcium-based agents increase the risk of hypercalcaemia and cardiovascular calcification. Consequently, there remains a need for new, safe and effective non-calcium-, non-aluminium-based alternative treatments. Fortunately, several new agents are now available or are in the late stages of development, including sevelamer hydrochloride and lanthanum carbonate. Although sevelamer hydrochloride represents a step forward in the management of hyperphosphataemia, it has several drawbacks and is far from being the ideal phosphate binder. Lanthanum carbonate is the most recent non calcium, non-aluminium phosphate binder to be developed for the treatment of hyperphosphataemia. Animal studies have shown it to be as effective as aluminium, without the associated toxicity. In clinical studies, lanthanum carbonate significantly reduced serum phosphorus levels, compared with placebo. It shows a similar efficacy to calcium carbonate in controlling serum phosphorus levels, but requires lower doses. In addition, lanthanum carbonate is at least as well tolerated as calcium carbonate, but is not associated with hypercalcaemia. Importantly, it has a positive effect on bone histology, with no evolution towards low bone turnover. Lanthanum carbonate, therefore, moves closer to the ideal phosphate binder. PMID- 15126651 TI - Quality improvement guidelines for recording patient radiation dose in the medical record. PMID- 15126652 TI - Percutaneous treatment of low flow vascular malformations. AB - Low flow vascular malformations, especially venous and macrocystic lymphatic malformations, are effectively treated by percutaneous intralesional injection of sclerosant drugs, such as ethanol and detergent sclerosant drugs. Good to excellent results are possible in 75%-90% of patients who undergo serial sclerotherapy. Most adverse effects are manageable, but severe complications can result from the intravascular administration of ethanol. It is generally recommended that the treatment of vascular malformations be performed in a multidisciplinary setting by practitioners with appropriate training and support. PMID- 15126653 TI - Comparison of transcatheter arterial embolization and surgery for treatment of bleeding peptic ulcer after endoscopic treatment failure. AB - PURPOSE: To compare the outcomes of embolotherapy and surgery as salvage therapy after therapeutic endoscopy failure in the treatment of upper gastrointestinal peptic ulcer bleeding. MATERIALS AND METHODS: Retrospective analysis of 70 cases of refractory peptic upper gastrointestinal hemorrhage was performed. Thirty-one cases were managed with embolotherapy and 39 were managed surgically. Demographic variables, underlying conditions, clinical findings, endoscopic treatment, transfusion requirements before and after alternative therapeutic approach, length of hospital stay, and outcomes including recurrent bleeding, need for surgery after initial alternative treatment, and in-hospital death were recorded. RESULTS: Patients who received embolotherapy were older (75.2 years +/- 10.9 vs 63.3 years +/- 14.5; P <.001) and had greater incidences of heart disease (67.7% vs 20.5%; P <.001) and previous anticoagulation treatment (25.8% vs 5.1%; P =.018). There were no differences in the rest of the pretreatment variables. No differences were found between the embolotherapy and surgery groups in the incidence of recurrent bleeding (29% vs 23.1%), need for additional surgery (16.1% vs 30.8%), or death (25.8% vs 20.5). CONCLUSIONS: The lack of differences between these two treatment alternatives, despite the more advanced age and greater prevalence of heart disease in the embolotherapy group, provides support for future prospective randomized studies aimed to evaluate the role of embolotherapy in the management of refractory peptic ulcer bleeding. PMID- 15126654 TI - Embolotherapy of pulmonary arteriovenous malformations: long-term results in 112 patients. AB - PURPOSE: To evaluate the long-term results of embolotherapy of pulmonary arteriovenous malformations (PAVMs) in a large group of patients. MATERIALS AND METHODS: Between July 1988 and August 2001, 134 consecutive patients underwent embolotherapy of PAVMs with feeding arteries larger than 3 mm or that had previously caused bleeding or systemic complications. The mean follow-up was 62.2 months. The primary endpoints of the study were the efficacy of embolotherapy, decrease in right-to-left shunt, and increase in partial arterial oxygen pressure (PaO(2)); the secondary endpoint was the prevalence of complications. Standard follow-up consisted of yearly history, chest radiography, and arterial blood gas measurement. RESULTS: Follow-up was available in 112 patients. Initially, 296 PAVMs were embolized in these patients. Nineteen patients (17%) underwent a second procedure and four patients underwent a third procedure because of recanalization of originally occluded feeding arteries (25 PAVMs, 8%) or interval enlargement of untreated PAVMs (53 PAVMs). In total, 349 PAVMs were embolized in 157 sessions. The mean diameter of occluded vessels was 4.7 mm. The long-term outcomes of embolotherapy were successful in 83% of patients overall and in 96% of patients in whom all angiographically visible PAVMs were embolized. Recanalization occurred in 12 of 16 patients who underwent repeat treatment because of enlargement of nonembolized PAVMs. Postprocedural pleurisy occurred after 14 of 157 sessions (9%). Periprocedural complications occurred in 12 sessions (8%) and included migration of an embolic device, transient ischemic attack (TIA), angina pectoris, and early cerebral infarction after embolization. Three patients experienced TIA and two patients experienced a cerebral abscess during follow-up after embolotherapy. CONCLUSIONS: Embolotherapy of PAVMs is efficacious and durable in the majority of patients. Patients should remain under regular review because recanalization of PAVMs or enlargement of untreated PAVMs can occur years after treatment. PMID- 15126655 TI - Retrospective assessment of risk factors to predict tunneled hemodialysis catheter outcome. AB - PURPOSE: To identify factors that may influence the function, outcome, and complications associated with tunneled hemodialysis catheters. MATERIALS AND METHODS: Radiology reports and hemodialysis, medical, and Clinical Information System (computerized patient medical record system) records were retrospectively reviewed in 221 consecutive patients who underwent tunneled hemodialysis catheter placement by interventional radiologists between January 11, 1996 and January 13, 2000 at Dartmouth-Hitchcock Medical Center. Various patient characteristics (diabetes, smoking, hypertension, age, sex, atherosclerotic cardiovascular disease, history of cardiac catheterization, coumadin use, functional status, and obesity) were assessed for their relationship to the outcome of hemodialysis catheters. Catheter outcome was examined by calculating infection rate, thrombosis rate, fibrin formation rate, mechanical malfunction rate, and total complication rate. With these patient characteristics and outcome variables, multiple regression analysis was performed with STATA (College Station, TX) statistical analysis software. RESULTS: Of the 221 patients reviewed, 39 patients were lost to follow-up. Among the remaining 182 patients, 427 catheters were placed for a total number of 36994 catheter-days. For overall complication rate, multiple regression analysis revealed a statistically significant positive correlation only for hypertension (P =.032). Total complication rate was 0.76 events per 100 catheter-days (95% CI: 0.53-1.00) for patients with a documented history of hypertension and 0.27 events per 100 catheter-days (95% CI: 0.08-0.45) for patients without (P =.024, paired student t test). For patients with diabetes versus patients without, the infection rates were 0.34 episodes per 100 catheter days (95% CI: 0.15-0.53) and 0.12 episodes per 100 catheter-days (95% CI: 0.06 0.18), respectively, (P =.011, paired student t test). Thrombosis rate for patients on coumadin was 0.13 events per 100 catheter-days (95% CI: -0.14-0.40), while thrombosis rate for patients not taking coumadin was 0.03 events per 100 catheter-days (95% CI: 0-0.05) (P =.036, paired student t test). CONCLUSION: Hypertension is a risk factor for poor outcome of tunneled hemodialysis catheters as measured by total complication rate requiring catheter removal or exchange. In this retrospective study, no other specific risk factors predicted an increased need for removal or exchange of tunneled hemodialysis catheters. PMID- 15126656 TI - Percutaneous radiofrequency ablation of lung neoplasms: initial therapeutic response. AB - PURPOSE: To evaluate the feasibility, safety, and initial therapeutic effect of radiofrequency (RF) ablation in the treatment of unresectable malignant lung tumors. MATERIALS AND METHODS: Fifty-four lung neoplasms in 31 patients were treated with RF ablation. Thirteen tumors were primary lung cancers and 41 were pulmonary metastases. Tumor sizes ranged from 0.7 to 6.0 cm, with a mean size of 2.7 +/- 1.3 cm. After the RF electrode was placed in the tumor with computed tomographic (CT) fluoroscopic guidance, RF energy was applied. Initial therapeutic response was evaluated by (18) F fluorodeoxyglucose positron emission tomography (FDG-PET) and contrast-enhanced CT. The disappearance of FDG uptake on PET images and tumor enhancement on CT images were considered to indicate complete tumor necrosis. Complete necrosis rates were evaluated according to tumor size and type (primary or secondary lung neoplasm). RESULTS: RF ablation was technically successful in all lesions. Complete necrosis was achieved in 32 of the 54 tumors (59%) after initial RF session. There was a significant difference in the rate of complete tumor necrosis between tumors 3 cm or less and tumors larger than 3 cm (69% vs. 39%; P <.05). Tumor type did not influence complete necrosis rates. Lung abscesses developed in two patients with large tumors. CONCLUSION: Lung RF ablation is a feasible, relatively safe, and promising treatment for unresectable lung neoplasms. Tumor size is an important factor in achieving complete tumor necrosis. PMID- 15126657 TI - Clinical outcomes with airway stents for proximal versus distal malignant tracheobronchial obstructions. AB - PURPOSE: This study evaluates the outcome of tracheobronchial stent placement in symptomatic patients with malignant disease. MATERIALS AND METHODS: From 1993 to 2002, 30 patients had stents placed for malignant strictures. Five of 30 patients underwent stent placement distal to the mainstem bronchi, 13 received stents in both the proximal and distal airways, and 12 received stents in only the proximal airways. Clinical response and survival were determined from the patients' medical records. A positive clinical response was judged to have occurred if the patient improved in two of these three categories: subjective symptoms (patient reported), objective signs (clinician-reported), and postprocedural imaging. RESULTS: The condition of 29 of 30 patients improved within 4 weeks of stent placement. The mean survival duration after stent placement was 261 days (SD, 395.1 days). The location of stent placement was not associated with significant differences in clinical improvement or survival (P =.51). Eight patients had additional airway segments that were too diffusely involved in which to place a stent or could not be recanalized. Mean survival in this group with incomplete stent placement was significantly reduced at 24.9 days (SD, 23.1 days), compared with 345.5 days (SD, 436 days) for the remaining patients who underwent complete stent placement (P <.05). Four patients lived less than 10 days after the procedure and three patients (75%) had mediastinal invasion. CONCLUSION: Tracheobronchial stent placement effectively palliates malignant airway obstruction, and clinical improvement is independent of the location in which the stent is implanted. When patients had diffuse or highly obstructive airway involvement and underwent incomplete stent placement, clinical response was also satisfactory, even though survival was worse. Patients with mediastinal invasion were poor candidates for stent placement as a result of their short survival. PMID- 15126658 TI - Factors associated with pneumothorax and pneumothorax requiring treatment after percutaneous lung biopsy in 443 consecutive patients. AB - PURPOSE: To describe patient- and procedure-related factors associated with post biopsy pneumothorax and those that require intervention. MATERIALS AND METHODS: Patient and procedure data from all lung biopsies performed at a single center between January 2000 and July 2001 were recorded prospectively. Data included patient demographics, lesion size, lesion depth from skin, needle size, number of passes, patient position during biopsy, imaging method used (computed tomography/fluoroscopy), if sedation was used, occurrence of pneumothorax and whether the pneumothorax required treatment. Patient charts were retrospectively reviewed to determine smoking history, as well as previous ipsilateral chest surgery or radiation therapy. Univariate and multivariate analysis was performed, and P <.05 was considered significant. RESULTS: Four-hundred fifty-three biopsies were performed on 443 patients. One-hundred six patients (23.4%) had post-biopsy pneumothorax and 31 patients (6.8% overall, 29.2% of pneumothorax group) required intervention. By univariate analysis, increased patient age, smaller lesion size, increased depth from skin, supine position, and no history of surgery were significant predictors of biopsy-related pneumothorax. However, only increased patient age, supine position, no history of ipsilateral surgery, and history of smoking were associated with pneumothorax that required intervention. By multivariate analysis, increased patient age, smaller lesion size, and no history of surgery predicted pneumothorax; supine position, history of smoking, and no history of ipsilateral surgery predicted which patients with pneumothorax would require treatment. CONCLUSION: Independent risk factors for pneumothorax include increased patient age, smaller lesion size, and no history of surgery. Previous surgery and prone positioning during biopsy appear to provide a "protective effect" against clinically significant post-biopsy pneumothorax. PMID- 15126659 TI - Hemodynamic effects of clot entrapment in the TrapEase inferior vena cava filter. AB - PURPOSE: The TrapEase vena cava filter has a symmetric design. Emboli can be trapped in the outlet conical section (superior cone) or between the filter and vessel wall at the inlet end (inferior cone). The purpose of this in vitro study is to investigate the hemodynamic effects of clot entrapment by the TrapEase filter and to examine the possibility of flow-induced filter thrombosis. MATERIALS AND METHODS: Velocity and wall shear stress maps were determined for steady flow with use of the photochromic flow visualization technique. Experiments were done for a filter without clot and for three other cases: an asymmetric clot in the inferior zone, a symmetric clot in the superior zone, and a filter with both zones partially occluded. Each simulated clot was 1500 mm(3) and the vessel diameter was 2 cm. RESULTS: The unoccluded filter did not significantly affect the flow field. However, for a partially occluded filter, flow stagnation/recirculation and turbulence developed downstream from the clot. The greatest effect was noted when the clot was against the vessel wall in the inferior trapping region. CONCLUSIONS: The tendency for clots to be trapped between the filter and the vessel wall in the inferior (inlet) region may play an important role in the performance of the TrapEase filter. A clot in this configuration will generate a large region of flow stagnation/recirculation that is considered to be prothrombotic. In addition, a significant amount of the filter wire will be embedded in this region, which may also promote thrombosis. PMID- 15126660 TI - Location of vessel occlusion of calibrated tris-acryl gelatin microspheres for tumor and arteriovenous malformation embolization. AB - PURPOSE: To evaluate (i). the presence and number of calibrated tris-acryl gelatin microspheres (TGMS) in targeted organs after embolization of tumors or arteriovenous malformations (AVMs) and (ii). the possible correlations among the size of TGMS used for embolization, the size of TGMS found in specimens, and the size of the occluded vessels. MATERIALS AND METHODS: Histologic slides were reviewed of 92 specimens from 80 patients with primarily head and neck tumors or AVMs operatively treated after embolization with TGMS of various sizes (40-120 microm, 100-300 microm, 300-500 microm, 500-700 microm, 700-900 microm, and 900 1200 microm). The diameters of the vessels containing TGMS, the size of TGMS, and the thickness of the inflammatory reactions developed around them were measured, and the location of the microspheres was recorded. RESULTS: TGMS were found in 88% of the specimens; 1985 embolized vessels containing TGMS were analyzed. The median number of TGMS per vessel was one (mean +/- SD, 3.1 +/- 6.9). The diameter of the occluded vessels increased significantly (P <.0001) with increased size of TGMS used for embolization. In tumors, 92% of the occluded vessels were located inside the tumor. CONCLUSION: There is an obvious correspondence between the size of the TGMS used for embolization and the diameter of the occluded vessels. This correlation confirms the possibility that the vessels to be occluded can be precisely targeted with the use of a proper TGMS size range. PMID- 15126661 TI - Delayed migration of a catheter fragment from the left to the right pulmonary artery. AB - A 45-year-old woman is described in whom an intrapulmonary venous access catheter fragment, originally left in place when discovered 1 year after port removal, migrated from one side to the other 2 years later. The patient underwent uncomplicated percutaneous removal of the fragment via the right internal jugular vein. The observed fragment mobility adds further argument for removing all intravenous catheter fragments, even if they have embolized to the pulmonary artery. PMID- 15126662 TI - Portal vein occlusion with aberrant left gastric vein functioning as a hepatopetal collateral pathway. AB - A rare case of an aberrant left gastric vein functioning as a hepatopetal collateral as a result of portal vein occlusion is presented herein. The portal venous phase of multislice computed tomography clearly demonstrated this anatomic variation and provided a reliable vascular "road map" for percutaneous transhepatic portal venous stent placement. Portal hypertension associated with extensive gastrojejunal varices improved dramatically after stent placement. PMID- 15126663 TI - Transportal intravariceal sclerotherapy with N-butyl-2-cyanoacrylate for gastric varices. AB - Four patients with gastric varices without catheterizable draining veins through a systemic vein were treated with transportal intravariceal sclerotherapy with n butyl-2-cyanoacrylate (NBCA). In all patients, the gastric varices were successfully obliterated with 2-5 mL of NBCA-lipiodol mixture injected via a microcatheter introduced into the varices with transhepatic or transileocolic portal venous access. No complications related to the procedure were encountered. Follow-up gastroendoscopy showed disappearance (n = 3) or marked decrease (n = 1) of the varices. Neither recurrent gastric varices nor variceal bleeding were observed during the follow-up period (6-46 months). Transportal intravariceal sclerotherapy is useful for obliteration of gastric varices in selected cases. PMID- 15126664 TI - Thrombolysis for central venous occlusion causing bilateral chylothorax in a patient with down syndrome. AB - A 20-year-old woman with Down syndrome (trisomy 21) and acute lymphoblastic leukemia presented with severe respiratory compromise secondary to bilateral chylothorax as a result of central venous thrombosis and extensive upper-limb deep venous thrombosis. The chylothorax was successfully managed by catheter directed thrombolysis and angioplasty of the venous occlusions. The development of venous thrombosis was likely to have been multifactorial. It is recognized that there is an increased incidence of congenital lymphatic anomalies in Down syndrome, which may have been a contributing factor in the development of chylothorax in this patient. This report illustrates the angiographic findings, demonstrates the successful vascular recanalization, and discusses the etiology and management of central venous thrombosis and chylothorax. The case is also presented to contribute to the expanding evidence in support of catheter-directed venous thrombolysis in selected clinical circumstances. PMID- 15126665 TI - High-dose intravenous gadolinium for renal computed tomographic angiography. PMID- 15126666 TI - Re: catheter-directed thrombolytic therapy for limb ischemia: current status and controversies. PMID- 15126667 TI - Reporting standards for carotid artery angioplasty and stent placement. PMID- 15126668 TI - Automated structure prediction of weakly homologous proteins on a genomic scale. AB - We have developed TASSER, a hierarchical approach to protein structure prediction that consists of template identification by threading, followed by tertiary structure assembly via the rearrangement of continuous template fragments guided by an optimized C(alpha) and side-chain-based potential driven by threading based, predicted tertiary restraints. TASSER was applied to a comprehensive benchmark set of 1,489 medium-sized proteins in the Protein Data Bank. With homologues excluded, in 927 cases, the templates identified by our threading algorithm PROSPECTOR_3 have a rms deviation from native <6.5 A with approximately 80% alignment coverage. After template reassembly, this number increases to 1,172. This shows significant and systematic improvement of the final models with respect to the initial template alignments. Furthermore, significant improvements in loop modeling are demonstrated. We then apply TASSER to the 1,360 medium-sized ORFs in the Escherichia coli genome; approximately 920 can be predicted with high accuracy based on confidence criteria established in the Protein Data Bank benchmark. These results from our unprecedented comprehensive folding benchmark on all protein categories provide a reliable basis for the application of TASSER to structural genomics, especially to proteins of low sequence identity to solved protein structures. PMID- 15126670 TI - Assessment of hand osteoarthritis: correlation between thermographic and radiographic methods. AB - OBJECTIVE: Anatomical stages of digital osteoarthritis (OA) have been characterized radiographically as progressing through sequential phases from normal to osteophyte formation, progressive loss of joint space, joint erosion and joint remodelling. Our study was designed to evaluate a physiological parameter, joint surface temperature, measured with computerized digital infrared thermal imaging, and its association with sequential stages of radiographic OA (rOA). METHODS: Thermograms, radiographs and digital photographs were taken of both hands of 91 subjects with nodal hand OA. Temperature measurements were made on digits 2-5 at distal interphalangeal (DIP) joints, proximal interphalangeal (PIP) joints and metacarpophalangeal (MCP) joints (2184 joints in total). We fitted a repeated measures ANCOVA model to analyse the effects of rOA on temperature, with handedness, joint group, digit and NSAID use as covariates. RESULTS: The reliability of the thermoscanning procedure was high (generalizability coefficient 0.899 for two scans performed 3 h apart). The mean joint temperature decreased with increasing rOA severity, defined by the Kellgren Lawrence (KL) scale. The mean temperature of KL0 joints was significantly different from that of each of the other KL grades (P or = 100 W (P < 0.05). IC increased progressively in CON during exercise but remained unchanged with CL. CONCLUSION: Obesity-related chest loading decreases lung volumes and increases the mechanical ventilatory constraints during exercise and is likely a critical factor in reducing exercise capacity in obesity. PMID- 15126711 TI - Resistance training alters plasma myostatin but not IGF-1 in healthy men. AB - PURPOSE: We determined and compared the magnitude of changes in resting plasma myostatin and IGF-1, muscle strength, and size in response to whole body or local muscle resistance training in healthy men. METHODS: Volunteers performed high intensity resistance exercise of major muscle groups of the whole body (N = 11), or of the elbow flexors only (N = 6), twice per week for 10 wk. Strength was assessed by elbow flexor one-repetition maximum (1-RM) and repetitions at 80% of 1-RM, muscle cross-sectional area by MRI, and plasma IGF-1 by RIA and myostatin by Western analyses, before and after the training program. RESULTS: In subjects of both groups, elbow flexor 1-RM and cross-sectional area increased (P = 0.05) by 30 +/- 8% (mean +/- SD) and 12 +/- 4%, respectively. Individual changes in myostatin ranged from 5.9 to -56.9%, with a mean decrease of 20 +/- 16%, whereas IGF-1 did not change from pre- to posttraining. There were no significant differences in any of the responses of the subjects between the two training programs. CONCLUSION: Myostatin may play a role in exercise-induced increases in muscle size, its circulating levels decreasing with resistance training in healthy men. Exercise of the whole body versus the elbow flexors alone did not provide a supplementary stimulus in altering resting plasma IGF-1 or myostatin, or in increasing muscle strength or size. Thus, by default, growth factor responses local to the muscle may be more important than circulating factors in contributing to muscle hypertrophy with resistance training. PMID- 15126712 TI - Cytokines and growth factors during and after a wrestling season in adolescent boys. AB - PURPOSE: Brief periods of aerobic exercise training lead to reductions, rather then the expected increases in circulating IGF-I. We hypothesized that intense exercise training in adolescents initially leads to simultaneous increases in proinflammatory cytokines and decreases in activity of the GH/IGF-I axis; and that as exercise training proceeds, levels of proinflammatory cytokines become reduced, and a rebound in IGF-I ensues leading to the higher IGF-I levels. METHOD: To test this, we evaluated the GH/IGF-I axis and levels of inflammatory cytokines (IL-6, TNF-alpha, IL-1beta, IL-1ra), body composition, and fitness in 13 healthy adolescent boys (mean age 15.9 +/- 0.3 yr) over the course of a high school wrestling season. Subjects were tested preseason, midseason (6 wk), peak season (12-14 wk), and 4 wk postseason. RESULTS: No significant weight loss was noted throughout the season. During the wrestling season (mid and peak) both total (P < 0.046) and free (P < 0.002) IGF-I levels decreased, whereas proinflammatory cytokines (IL-1ra, P < 0.005; IGFBP-1, P < 0.013; and IGFBP-2, P < 0.025) increased. GHBP (P < 0.018) levels also decreased during the season. In the postseason, there were significant increases in GHBP, and free and total IGF I, whereas proinflammatory cytokines decreased. CONCLUSIONS: An initial catabolic type hormonal response occurs with intense exercise training in adolescents. This is followed by a rebound in circulating growth factors when the period of heavy training ceases. PMID- 15126713 TI - Increased VLDL-TAG turnover during and after acute moderate-intensity exercise. AB - PURPOSE AND METHODS: Breakdown rates of very low density lipoprotein triacylglycerols (VLDL-TAG) were quantified before (3 h), during (45 min), and after (3 h) moderate physical exercise at 40% VO2 max in young sedentary subjects (four male and four female, age 29.8 +/-1.6 yr, BMI 24.1 +/- 0.9 kg x m, VO2max 37.0 +/- 1.7 mL x kg x min), using boluses of H5-glycerol (100 mu mol x kg) and H2-palmitate (6.6 mu mol x kg). The catabolic rates of VLDL-TAG were calculated from the decay in the isotopic enrichment using a single-pool model. The results were compared with those obtained during 6 h of rest in five of the same volunteers. RESULTS: VLDL-TAG concentration remained constant throughout the study with exercise (P = NS). The fractional catabolic rate was nearly doubled from rest to exercise (P < 0.05 vs rest) and increased to an even greater extent during the early phase of recovery (P < 0.001 vs rest). During the late recovery phase, the value returned to the preexercise value (P = NS vs rest). The values for the absolute catabolic rate (kabs) followed the same trend. CONCLUSION: VLDL TAG turnover was increased during exercise and during the early phase of recovery. PMID- 15126714 TI - Induced metabolic alkalosis affects muscle metabolism and repeated-sprint ability. AB - PURPOSE: The purpose of this study was to assess the effects of induced metabolic alkalosis, via sodium bicarbonate (NaHCO3) ingestion, on muscle metabolism and power output during repeated short-duration cycle sprints. METHODS: : Ten active females (mean +/- SD: age = 19 +/- 2 yr, VO2max = 41.0 +/- 8.8 mL x kg x min ) ingested either 0.3 g x kg NaHCO3 or 0.207 g x kg of NaCl (CON), in a double blind, random, counterbalanced order, 90 min before performing a repeated-sprint ability (RSA) test (5 x 6-s all-out cycle sprints every 30 s). RESULTS: Compared with CON, there was a significant increase in resting blood bicarbonate concentration [HCO3] (23.6 +/- 1.1 vs 30.0 +/- 3.0 mmol x L ) and pH (7.42 +/- 0.02 vs 7.50 +/- 0.04), but no significant difference in resting lactate concentration [La] (0.8 +/- 0.2 vs 0.8 +/- 0.3 mmol x L ) during the NaHCO3 trial. Muscle biopsies revealed no significant difference in resting muscle [La], pH, or buffer capacity (beta(in vitro)) between trials (P > 0.05). Compared with CON, the NaHCO3 trial resulted in a significant increase in total work (15.7 +/- 3.0 vs 16.5 +/- 3.1 kJ) and a significant improvement in work and power output in sprints 3, 4, and 5. Despite no significant difference in posttest muscle pH between conditions, the NaHCO3 trial resulted in significantly greater posttest muscle [La]. CONCLUSIONS: As NaHCO3 ingestion does not increase resting muscle pH or beta(in vitro), it is likely that the improved performance is a result of the greater extracellular buffer concentration increasing H efflux from the muscles into the blood. The significant increase in posttest muscle [La] in NaHCO3 suggests that an increased anaerobic energy contribution is one mechanism by which NaHCO3 ingestion improved RSA. PMID- 15126715 TI - Conjugated linoleic acid improves insulin sensitivity in young, sedentary humans. AB - BACKGROUND: Preliminary evidence in obese diabetic rats suggests that conjugated linoleic acid (CLA) may have antidiabetic properties, based on reductions in fasting glucose and insulin concentrations. However, in lean rats, CLA causes hyperinsulinemia. Furthermore, experiments in humans also suggest that CLA may worsen insulin sensitivity. OBJECTIVES: The present study examined whether CLA supplementation can improve insulin sensitivity in humans. DESIGN: : Sixteen young sedentary individuals (age, 21.5 +/- 0.4 yr (mean +/- SEM); body mass, 77.6 +/- 3.4 kg) participated in this study. Ten subjects received 4 g x d of mixed CLA isomers (35.5%cis-9, trans-11; 36.8%trans-10, cis-12) for 8 wk, whereas six subjects received placebo (safflower oil). Oral glucose tolerance tests were performed at baseline (0), 4 and 8 wk of supplementation. RESULTS: : After 8 wk of CLA supplementation, insulin sensitivity index (ISI) increased (14.4 +/- 1.0, 8 wk vs 11.3 +/- 1.3, 0 wk; P < 0.05), which corresponded to a decrease in fasting insulin concentrations. Six of the 10 subjects showed large increases in their ISI (range, +27 to 90%), whereas two demonstrated essential no change (+3 to 5%), and two had a decrease in insulin sensitivity (-12 to -13%). ISI was unchanged over 8 wk in the placebo group. CONCLUSIONS: Our results indicate that a common dosage of a commercially available CLA supplement can improve ISI in young, sedentary individuals. However, there is considerable individual variability in the response. Additional studies are required to identify underlying metabolic changes in human skeletal muscle. PMID- 15126717 TI - Ground/foot impacts: measurement, attenuation, and consequences. PMID- 15126716 TI - Effects of running distance and training on Ca2+ content and damage in human muscle. AB - PURPOSE: Muscle damage and soreness are well-known adverse effects of running, especially when covering distances in excess of habitual running activity. Loss of Ca homeostasis is hypothesized to initiate the development of exercise-induced muscle damage. We tested the hypothesis that the Ca content of vastus lateralis muscle increases after a 10- or 20-km run and studied the relations between Ca accumulation and running distance, endurance training, and fiber type distribution. METHODS: Twenty-four healthy young men and women were divided into two groups who ran either 10 or 20 km. Muscle biopsies and blood samples were collected before, immediately after, and in the days after the run. RESULTS: : The Ca content in muscle biopsies increased from 0.70 +/- 0.02 to 0.93 +/- 0.04 micromol x g wet weight after the 20-km run (P < 0.001) and was still significantly elevated at 4 and 48 h after the run. In the 10-km runners, however, no significant increase in Ca was found (0.81 +/- 0.03 vs 0.91 +/- 0.06 micromol x g wet weight, P = 0.08). Plasma levels of lactate dehydrogenase and creatine kinase increased after both running distances, the increase being greatest after the 20-km run. Eight of the 10-km runners completed an endurance training program and subsequently repeated the 10-km run. The response to a new 10-km run with regard to muscle Ca content and parameters of muscle damage was essentially unchanged by training. CONCLUSIONS: The degree of muscle damage depends on running distance, and a significant Ca accumulation in muscle is seen after 20 km. Ten weeks of endurance training does not influence Ca homeostasis and muscle damage after 10-km running. PMID- 15126718 TI - The effects of knee contact angle on impact forces and accelerations. AB - This article will summarize findings from several studies that together allow: 1) the examination of the effect that knee contact angle has on the severity of the resulting impact, 2) examination of the relationship between vertical ground reaction impact forces and leg impact accelerations, and 3) exploration of the adaptations that occur in response to running during changing environmental conditions. Changing the knee flexion angle at contact can alter the effective mass during activities in which the foot impacts the ground. It has been shown that increasing the knee flexion angle at ground contact can reduce the peak vertical ground reaction impact force, but it can also increase the peak impact acceleration at the leg. Attenuation can be calculated from accelerometers on the leg and the head and combined with the leg acceleration values to give a more accurate impression of the severity of the impact. Lower-extremity joint contact angles can be used to examine the kinematic adaptations that take place in response to changing environmental conditions. One common adaptation that can occur when the internal or external environment is not ideal is an increase in the knee flexion angle at contact. More extended knee contact angles can increase the forces experienced by the body and therefore increase injury potential. Increased knee flexion may give the runner a larger margin for dealing with kinematic errors but this benefit likely has an associated metabolic cost that will reduce performance. PMID- 15126719 TI - Kinematic adaptations during running: effects of footwear, surface, and duration. AB - Repetitive impacts encountered during locomotion may be modified by footwear and/or surface. Changes in kinematics may occur either as a direct response to altered mechanical conditions or over time as active adaptations. PURPOSE: : To investigate how midsole hardness, surface stiffness, and running duration influence running kinematics. METHODS: In the first of two experiments, 12 males ran at metabolic steady state under six conditions; combinations of midsole hardness (40 Shore A, 70 Shore A), and surface stiffness (100 kN x m, 200 kN x m, and 350 kN x m). In the second experiment, 10 males ran for 30 min on a 12% downhill grade. In both experiments, subjects ran at 3.4 m x s on a treadmill while 2-D hip, knee, and ankle kinematics were determined using high-speed videography (200 Hz). Oxygen cost and heart rate data were also collected. Kinematic adaptations to midsole, surface, and running time were studied. RESULTS: Stance time, stride cycle time, and maximal knee flexion were invariant across conditions in each experiment. Increased midsole hardness resulted in greater peak ankle dorsiflexion velocity (P = 0.0005). Increased surface stiffness resulted in decreased hip and knee flexion at contact, reduced maximal hip flexion, and increased peak angular velocities of the hip, knee, and ankle. Over time, hip flexion at contact decreased, plantarflexion at toe-off increased, and peak dorsiflexion and plantarflexion velocity increased. CONCLUSION: Lower extremity kinematics adapted to increased midsole hardness, surface stiffness, and running duration. Changes in limb posture at impact were interpreted as active adaptations that compensate for passive mechanical effects. The adaptations appeared to have the goal of minimizing metabolic cost at the expense of increased exposure to impact shock. PMID- 15126720 TI - Impact and overuse injuries in runners. AB - Forces that are repeatedly applied to the body could lead to positive remodeling of a structure if the forces fall below the tensile limit of the structure and if sufficient time is provided between force applications. On the other hand, an overuse injury could result if there is inadequate rest time between applied forces. Running is one of the most widespread activities during which overuse injuries of the lower extremity occur. The purpose of this article is to review the current state of knowledge related to overuse running injuries, with a particular emphasis on the effect of impact forces. Recent research has suggested that runners who exhibit relatively large and rapid impact forces while running are at an increased risk of developing an overuse injury of the lower extremity. Modifications in training programs could help an injured runner return to running with decreased rehabilitation time, but it would be preferable to be able to advise a runner regarding injury potential before undertaking a running program. One of the goals of future research should be to focus on the prevention or early intervention of running injuries. This goal could be accomplished if some easily administered tests could be found which would predict the level of risk that a runner may encounter at various levels of training intensity, duration, and frequency. The development of such a screening process may assist medical practitioners in identifying runners who are at a high risk of overuse injury. PMID- 15126721 TI - Reliability of biomechanical variables of sprint running. AB - PURPOSE: The purpose of this paper was to report the reliability of variables used in the biomechanical assessment of sprint running and to document how these reliability measures are likely to improve when using the average score of multiple trials. METHODS: Twenty-eight male athletes performed maximal-effort sprints. Video and ground reaction force data were collected at the 16-m mark. The reliability (systematic bias, random error, and retest correlation) for a single score was calculated for 26 kinematic and 7 kinetic variables. In addition, the reliability (random error and retest correlation) for the average score of 2, 3, 4, and 5 trials was predicted from the reliability of a single score. RESULTS: For all variables, there was no evidence of systematic bias. The measures of random error and retest correlation differed widely among the variables. Variables describing horizontal velocity of the body's center of mass were the most reliable, whereas variables based on vertical displacement of the body's center of mass or braking ground reaction force were the least reliable. For all variables, reliability improved notably when the average score of multiple trials was the measurement of interest. CONCLUSION: Although it is up to the researcher to judge whether a measurement is reliable enough for its intended use, some of the lower-reliability variables were possibly too unreliable to monitor small changes in an athlete's performance. Nonetheless, there was a consistent trend for reliability to improve notably when the average score of multiple trials was the measurement of interest. Subsequently, if resources permit, researchers and applied sports-scientists may like to consider using the average score of multiple trials to gain the advantages that improved reliability offers. PMID- 15126722 TI - Selective activation of tibialis posterior: evaluation by magnetic resonance imaging. AB - PURPOSE: To determine which exercise most selectively and effectively activates tibialis posterior. METHODS: Five healthy adults (two men, three women; mean age 31 yr) with an Arch Index (AI) within 1 SD from norm performed three exercises, separated by 1-wk intervals. The exercises were: 1) closed chain resisted foot adduction (foot adduction), 2) unilateral heel raise (heel raise), and 3) open chain resisted foot supination (foot supination). Magnetic resonance transaxial images were obtained immediately before and after exercise using a 1.5-T MRI system. Changes in pre-to postexercise signal intensity were compared across five muscles: tibialis posterior, tibialis anterior, medial gastrocnemius, soleus, and peroneus longus. Postexercise signal intensity was normalized to baseline preexercise signal intensity. RESULTS: Tibialis posterior signal intensity increased after each exercise. The greatest TP increase (50 +/- 6%) occurred after foot adduction, whereas the mean increase in the other muscles was less than 5%. After the heel raise exercise, the signal intensity increase in TP was 27% (+/- 11%), soleus 39% (+/- 8%), peroneus longus 57% (+/- 14%), and medial gastrocnemius 99% (+/- 12%). The signal intensity of tibialis anterior decreased 4% (+/- 2%). After foot supination, the TP signal intensity increased 26% (+/- 7%), whereas the mean change in the other muscles was less than 10%. Multivariate analyses of variance revealed a significant difference in muscle activation between exercises. Posthoc analysis showed greater activation of TP during foot adduction than foot supination (P = 0.021). CONCLUSION: In individuals with a normal AI, TP was activated selectively and most effectively during foot adduction. Knowledge of selective activation of a muscle is necessary to provide an optimal environment for muscle strengthening and/or tendon rehabilitation. PMID- 15126723 TI - Factors influencing the exercise behavior of adults with physical disabilities. AB - PURPOSE: Examine the theorized associations of transtheoretical model (TTM) of behavior change constructs (behavioral and cognitive processes of change, decisional balance, and self-efficacy), along with exercise barriers, by stage of change for exercise behavior among individuals with physical disabilities. METHODS: Cross-sectional survey of 322 adults with physical disabilities residing in the United States. Most participants (84.3%) used some form of assistive device (e.g., artificial limb, wheelchair), were female (62.1%), and Caucasian (91.9%). The mean age of participants was 52.5 yr (SD = 13.9). Participants completed and returned questionnaires for each TTM construct, along with an exercise barriers measure. RESULTS: In univariate analyses, all constructs were significantly (P < 0.001) associated with the stages of change for exercise behavior, except for dramatic relief (P > 0.005). The largest portion of variance was derived from counter conditioning (eta = 0.45), followed by the five behavioral processes of change combined (eta = 0.40), self-liberation (eta = 0.31), self-efficacy (eta = 0.30), and self-reevaluation (eta = 0.28). Direct discriminant function (multivariate) analysis revealed four discriminant functions which accounted for 71.0% (P < 0.001), 20.9% (P < 0.001), 6.3% (P < 0.05), and 1.8% (P = 0.62), respectively, of the between-group (stage of change) variability. The overall stage of change classification accuracy was 70.8%. CONCLUSION: This was the first study to examine the stages of change for exercise behavior among adults with physical disabilities on the basis of the full TTM and exercise barriers. Overall, the results are in general agreement with existing evidence among nondisabled populations. This provides further cross-sectional support for the internal and external validity of TTM and exercise barriers among a unique and understudied population segment. PMID- 15126724 TI - School-level intraclass correlation for physical activity in adolescent girls. AB - PURPOSE: The Trial for Activity in Adolescent Girls (TAAG) is a multi-center group-randomized trial to reduce the usual decline in moderate to vigorous physical activity (MVPA) among middle-school girls. In group-randomized trials, the group-level intraclass correlation (ICC) has a strong inverse relationship to power and a good estimate of ICC is needed to determine sample size. As a result, we conducted a substudy to estimate the school-level ICC for intensity-weighted minutes of MVPA measured using an accelerometer. METHODS: To estimate the ICC, each of six sites recruited two schools and randomly selected 45 eighth grade girls from each school; 80.7% participated. Each girl wore an Actigraph accelerometer for 7 d. Readings above 1500 counts per half minute were counted as MVPA. These counts were converted into metabolic equivalents (MET) and summed over 6 a.m. to midnight to provide MET-minutes per 18-h day of MVPA. Minutes of MVPA per 18-h day also were calculated ignoring the MET value. RESULTS: The unadjusted school-level ICC for minutes of MVPA was 0.0205 (95%CI: -0.0079, 0.1727) and for MET-minutes of MVPA was 0.0045 (95% CI: -0.0147, 0.1145). Adjustment for age and BMI had no measurable effect, whereas adjustment for ethnicity reduced both ICC; adjusted values were 0.0175 (95% CI: -0.0092, 0.1622) for minutes of MVPA and 0.0000 (95% CI: -0.0166, 0.0968) for MET-minutes of MVPA. This information was used to calculate the number of schools and girls needed for TAAG to have 90% power to detect a 50% reduction in the decline of MET-minutes of MVPA between sixth and eighth grade. CONCLUSIONS: The results called for 36 schools in TAAG, with 120 girls invited for measurements at each school, and a minimum participation rate of 80%. PMID- 15126725 TI - Improving energy expenditure estimation for physical activity. AB - PURPOSE: The purpose of this study was to validate the Intelligent Device for Energy Expenditure and Activity (IDEEA) for estimation of energy expenditure during a variety of activities. An additional aim was to improve the accuracy of the estimation of energy expenditure of physical activity based on second-by second information of type, onset, and duration of activity. METHODS: This study included two tests: a mask calorimetry test with 27 subjects [age = 33.7 +/- 13.8 (mean +/- SD) yr; BMI = 24.8 +/- 4.8 kg x m] and a respiratory chamber calorimetry test with 10 subjects (age = 32.9 +/- 12.4 yr; BMI = 26.1 +/- 5.6 kg x m). In the mask test, the subjects performed activities (sitting, standing, lying down, level treadmill walking, and running at different speeds) for 50-min durations. For the chamber test, subjects lived in the metabolic chamber for 23 h and performed three exercise sessions to compensate for the confined environment. RESULTS: The results showed significant correlations (P < 0.0001) between energy expenditure estimated by IDEEA and energy expenditure measured by the calorimeters with an accuracy >95%. After corrections for the decrease in sleeping metabolic rate, the estimation accuracy for the chamber test was increased by 1-96.2%, whereas the estimation accuracy for nighttime activity was significantly improved by 4-99%. CONCLUSION: IDEEA provides a suitable method for estimating the energy expenditure of physical activity. It provides both instantaneous and cumulative estimates of energy expenditure over a given period. PMID- 15126726 TI - Associations between physical activity dose and health-related quality of life. AB - PURPOSE: Although the beneficial effects of participation in regular physical activity (PA) are widely accepted, dose-response relationships between PA and health-related quality of life (HRQOL) remain unclear. METHODS: We examined relationships between frequency, duration, and intensity of PA and HRQOL among 175,850 adults using data from the 2001 BRFSS. Logistic regression was used to obtain odds ratios (OR) and 95% confidence intervals (CI) adjusted for age, gender, race/ethnicity, education, smoking status, and body mass index. RESULTS: The age-standardized prevalence (standard error) of 14 or more unhealthy (physical or mental) days during the previous 30 d was 28.4% (0.50) among physically inactive adults, 16.7% (0.27) among those with insufficient levels of PA, and 14.7% (0.22) among adults who met recommended levels. Overall, participation in no moderate PA (OR: 2.02; 95% CI: 1.85-2.21) was associated with an increased likelihood of having 14 or more unhealthy days. Also for moderate PA, participation every day of the week (5-6 d x wk as referent) (OR: 1.35; 1.26 1.46) was associated with an increased likelihood of 14 or more unhealthy days, as was participation for periods < 20 min (OR: 1.43; 95% CI: 1.30-1.58) or > or = 90 min (OR: 1.22; 95% CI: 1.14-1.31) per day (30-59 min x d as referent). Similar associations were observed for participation in vigorous PA. CONCLUSION: Persons achieving recommended levels of PA were more likely to report fewer unhealthy days compared with inactive and insufficiently active persons; however, participation in daily moderate or vigorous PA and participation in very short (< 20 min x d) or extended ( > or = 90 min x d) periods of PA was associated with poorer HRQOL. Further research examining the relationship between the dose of PA and HRQOL as well as other health outcomes is needed. PMID- 15126727 TI - Evaluation of the SenseWear Pro Armband to assess energy expenditure during exercise. AB - PURPOSE: To assess the accuracy of the SenseWear Pro Armband for estimating energy expenditure during exercise. METHODS: : Forty subjects (age = 23.2 +/- 3.8 yr; body mass index = 23.8 +/- 3.1 kg x m) performed four exercises (walking, cycling, stepping, arm ergometry) with each exercise lasting 20-30 min and workload increasing at 10-min intervals. Subjects wore the SenseWear Pro Armband on the right arm, and energy expenditure was estimated using proprietary equations developed by the manufacturer. Estimated energy expenditure from the SenseWear Pro Armband was compared with energy expenditure determined from indirect open-circuit calorimetry, which served as the criterion measure. RESULTS: : When a generalized proprietary algorithm was applied to the data, the SenseWear Pro Armband significantly underestimated total energy expenditure by 14.9 +/- 17.5 kcal (6.9 +/- 8.5%) during walking exercise, 32.4 +/- 18.8 kcal (28.9 +/- 13.5%) during cycle ergometry, 28.2 +/- 20.3 kcal (17.7 +/- 11.8%) during stepping exercise, and overestimated total energy expenditure by 21.7 +/- 8.7 kcal (29.3 +/- 13.8%) during arm ergometer exercise (P < or = 0.001). At the request of the investigators, exercise-specific algorithms were developed by the manufacturer and applied to the data that resulted in nonsignificant differences in total energy expenditure between indirect calorimetry and the SenseWear Pro Armband of 4.6 +/- 18.1 kcal (2.8 +/- 9.4%), 0.3 +/- 11.3 kcal (0.9 +/- 10.7%), 2.5 +/- 18.3 kcal (0.9 +/- 11.9%), and 3.2 +/- 8.1 kcal (3.8 +/- 9.9%) for the walk, cycle ergometer, step, and arm ergometer exercises, respectively. CONCLUSIONS: It appears that it is necessary to apply exercise-specific algorithms to the SenseWear Pro Armband to enhance the accuracy of estimating energy expenditure during periods of exercise. When exercise-specific algorithms are used, the SenseWear Pro Armband provides an accurate estimate of energy expenditure when compared to indirect calorimetry during exercise periods examined in this study. PMID- 15126728 TI - Motion sensor accuracy under controlled and free-living conditions. AB - PURPOSE: Two studies were conducted to examine the concurrent accuracy of the Yamax SW-200 (YAM), Omron HJ-105 (OM), and Sportline 330 (SL) pedometers, as well as a CSA accelerometer. METHODS: In study 1, motion sensor performance was evaluated against actual (observed) steps taken during 5-min bouts at five different treadmill speeds (54, 67, 80, 94, and 107 m x min) using a two-way repeated measures ANOVA (instrument x speed). Additionally, the direction and magnitude of motion sensor error was examined. In study 2, pedometer performance during 24 h of free-living was evaluated against the steps detected by the CSA criterion. The direction and magnitude of pedometer error was also examined in the free-living condition. RESULTS: In study 1, the SL showed significant differences from actual steps taken at all treadmill speeds (P < 0.05). Further, the absolute value of percent error was greatest for the SL at all treadmill speeds. At the slowest treadmill speed (54 m x min), the absolute value of percent error increased for the YAM and OM. In study 2, only the SL detected fewer steps than the CSA criterion (P < 0.05). The YAM demonstrated the lowest absolute value of percent error under free-living conditions. CONCLUSIONS: Different brands of motion sensors detect steps differently; therefore, caution must be used when comparing step counts between studies that have employed different brands of motion sensors. Taking into consideration the results of both studies and the initial walking test used for instrument screening purposes, it appears that, of the three pedometers tested, the YAM pedometer is most consistently accurate under both controlled and free-living conditions. Future research must consider presenting motion sensor accuracy in absolute terms so that the magnitude of error is not underestimated. PMID- 15126729 TI - Relationship between accumulated walking and body composition in middle-aged women. AB - PURPOSE: The purpose of this study was to examine the relationship between objectively determined physical activity (pedometer counted steps per day) and body composition variables in middle-aged women. METHODS: Height, weight, body fat percentage (%BF), waist circumference, and hip circumference were measured on eighty women (50.3 +/- 6.8 yr). For 7 d after testing, each subject wore a pedometer throughout the day while following her normal daily routine. Each morning the pedometer was reset to zero, and each evening the subject recorded the steps accumulated during the day. Pearson product moment correlations were used to examine the relationship between average steps per day and body composition variables. Subjects were placed in groups to reflect different levels of physical activity: inactive (<6000 steps x d), somewhat active (6000-9999 steps x d), and regularly active (> or = 10,000 steps x d). ANOVA was utilized to determine whether body composition variables varied across activity groups. Significance was set at P < 0.05 for all tests. RESULTS: : A significant correlation was found between average steps per day and %BF (-0.713, P < 0.0001); body mass index (BMI) (-0.417, P < 0.0001); waist circumference (-0.616; P < 0.0001); hip circumference (-0.278; P = 0.013); and waist-to-hip ratio (-0.652; P < 0.0001). There was a significant difference in body composition variables among activity groups, with higher values found in the less active groups. CONCLUSION: This is the first study to specifically examine the relationship between steps per day and body composition in middle-aged women. Although the cross-sectional nature of this study does not allow causal relationships to be determined, women who walked more had lower %BF. Additionally, the average BMI of women who accumulated 10,000+ steps x d was in the normal range. PMID- 15126730 TI - A need for more experimental studies of physical activity during childhood. PMID- 15126732 TI - Clinical predictors in obstructive sleep apnea patients with computer-assisted quantitative videoendoscopic upper airway analysis. AB - AIM: To identify the clinical predictors and assist surgeons in their clinical management of obstructive sleep apnea (OSA). A prospective study with a new approach to analyze the static and dynamic upper airway morphology between patients with OSA and normal subjects. METHOD: Quantitative computer-assisted videoendoscopy (validated with upper airway magnetic resonance imaging) was performed in 49 (43 males, 6 females) patients with OSA and compared with 39 (22 males, 17 females) controls (apnea-hypopnea index [AHI] < 5). Absolute cross sectional areas and transverse and longitudinal diameters at the retropalatal and retrolingual levels were measured during end of quiet respiration and during Mueller's maneuver in the erect and supine positions, allowing us to study static and dynamic morphology (collapsibility) of the upper airway. Three thousand seven hundred forty-four (3,744) parameters were analyzed. RESULTS: In males, retropalatal and retrolingual areas during Mueller's maneuver in the supine position of 0.7981 cm (relative operating characteristics [ROC] = 0.9284, positive pressure ventilation [PPV] = 86.05%, negative pressure ventilation [NPV] = 84.62%) and 2.0648 cm (ROC = 0.8183, PPV = 76%, NPV = 83.33%), respectively, were found to be good predictors/cut-off values for OSA. The retropalatal area measured in the supine position (AS1 mol/L) and collapsibility of the retropalatal area in the supine position (CAS1) were found to have significant correlations with severity of OSA. In females, the areas measured during Mueller's maneuver in the supine position of 0.522 cm at the retropalatal level (ROC = 1, 100% PPV and NPV) and the transverse diameter at the retrolingual level during erect Mueller's maneuver of 1.1843 cm (ROC = 0.9056, PPV = 100%, NPV = 83.33%) were found to be predictive. All measurements at the retropalatal level and in the supine position had higher predictability. Area measurements obtained during Muller's maneuver were more predictive (ROC > 0.9910) than resting measurements (ROC >0.8371). Several sex and anatomic-site specific formulas with excellent predictability (ROC close or equal to 1) were also devised. CONCLUSION: Upper airway Mueller's studies are predictive and useful (independent samples t test/Mann-Whitney U test, ROC) in identifying patients with OSA. With these sex and anatomic-site specific OSA predictors/formulas and this innovative clinical method, we hope to assist other surgeons with quantitative clinical diagnosis, assessment, surgical planning, and outcome assessment tools for OSA patients. PMID- 15126733 TI - Diagnosis and management of spontaneous cerebrospinal fluid-middle ear effusion and otorrhea. AB - OBJECTIVES/HYPOTHESIS: Spontaneous leak of cerebrospinal fluid (CSF) into the middle ear can occur in adults without a history of temporal bone trauma or fracture, meningitis, or any obvious cause. Therefore, clues may be lacking that would alert the otolaryngologist that fluid medial to an intact eardrum, or fluid emanating from an eardrum perforation, is likely to be CSF fluid. A review of relevant medical literature reveals that herniation of the arachnoid membrane through a tegmen defect may be congenital, or CSF leak may occur when dynamic factors (i.e., brain pulsations or increases in intracranial pressure) produce a rent in the arachnoid membrane. Because tegmen defects may be multiple rather than single, identifying only one defect may not be sufficient for achieving definitive repair. Data on nine cases of spontaneous CSF leak to the ear in adult patients from four medical centers are presented and analyzed to provide collective information about a disorder that can be difficult to diagnose and manage. STUDY DESIGN: Retrospective review of nine cases of spontaneous CSF middle ear effusion/otorrhea. RESULTS: The majority of patients presented with symptoms of aural fullness and middle ear effusion. Many developed suspicious clear otorrhea only after insertion of a tympanostomy tube. Two patients had multiple defects in the tegmen and dura, and five patients had meningoencephaloceles confirmed intraoperatively. Five patients underwent combined middle cranial fossa/transmastoid repair. Materials used in repair included temporalis fascia, free muscle graft, Oxycel cotton, calvarial bone, pericranium, bone wax, and fibrin glue. CONCLUSIONS: CSF middle ear effusion/otorrhea can develop in adults without a prior history of meningitis or head trauma or any apparent proximate cause. Although presenting symptoms can be subtle, early suspicion and confirmatory imaging aid in establishing the diagnosis. Because surgical repair by way of a mastoid approach alone can be inadequate if there are multiple tegmen defects, a middle fossa approach alone, or in combination with a transmastoid approach, should be considered in most cases. PMID- 15126734 TI - Quality of life in treatment of acute rhinosinusitis with clarithromycin and amoxicillin/clavulanate. AB - OBJECTIVES: Clarithromycin and amoxicillin/clavulanate (A/C) are first line antibiotics used to treat uncomplicated acute rhinosinusitis (ARS). This study examined the efficacy of clarithromycin and A/C for the treatment of ARS relative to the patient's quality of life (QOL). STUDY DESIGN: Twenty-two patients with uncomplicated ARS were randomly assigned treatment (single blinded relative to the investigator) using clarithromycin or A/C. Patients underwent assessment to confirm the diagnosis and treatment outcome at the initial screening and on completion of antibiotics (diagnosis + 14 days and 28 days). QOL was evaluated using the Allergy Outcomes Survey (AOS), the Rhinoconjunctivitis QOL Questionnaire (RQLQ), the Short Form 36 survey (SF-36), an instantaneous six-item Symptom Severity Survey (SSS-6), and a Visual Analogue Scale (VAS). Surveys were completed at the time of diagnosis, on completion of antibiotics, and at 28 days after diagnosis. RESULTS: Twenty patients completed the study. The SSS-6 and the RQLQ demonstrated significant improvement for all patients at week 4 (P =.002 and P =.003, respectively). The SSS-6 demonstrated significant improvement for clarithromycin at 14 days (P =.02) and at 28 days (P =.029), whereas A/C patients demonstrated significant improvement in symptoms only at 28 days (P =.046). The RQLQ, which reflects the previous 2 weeks, demonstrated significant improvement for the A/C patients at 28 days (P =.01). The Allergy Survey, the SF-36, and the VAS failed to demonstrate significant improvement in the combined data analysis. CONCLUSIONS: Clarithromycin and A/C were equally effective in treating ARS. The clarithromycin patients felt better more rapidly (at 14 days), but both groups of patients had long-term improvement in symptoms at 28 days. PMID- 15126735 TI - Recurrence rates after endoscopic sinus surgery for massive sinus polyposis. AB - BACKGROUND AND OBJECTIVES: Most studies on outcome after endoscopic sinus surgery (ESS) include patients with varying degrees of disease severity. Recurrence rates cited by those studies may not apply to the subset of patients with severe polyposis. Our aim is to provide reference information for recurrence rates and need for revision surgery in patients with severe disease. STUDY DESIGN, PATIENTS, AND METHODS: Review of patients with severe polyposis with a minimum Lund-McKay score of 16 and with a Kennedy computed tomography stage 3 or 4. Data collection included demographics, presence of asthma or documented allergy, history of previous surgery, extent of surgery, preoperative and postoperative management, recurrence rates, revision surgery rates, and follow-up. RESULTS: One hundred and eighteen records were reviewed. Fifty-nine (50%) patients had asthma, and 93 (79%) had documented allergy. All patients required extensive bilateral nasal polypectomy, complete anterior and posterior ethmoidectomy, and maxillary sinusotomy. One hundred (85%) also had frontal or sphenoid sinusotomy. Follow-up ranged from 12 to 168 (median 40) months. Seventy-one (60%) developed recurrent polyposis. Fifty-five (47%) were advised to undergo revision surgery, and 32 (27%) underwent surgery. History of previous sinus surgery or asthma predicted higher recurrence (P <.005, P <.001) and revision surgery rates (P =.02, P <.001). History of allergy also predicted recurrence and need for revision (P <.001, P <.001). CONCLUSIONS: Recurrence rates after ESS for severe polyposis are significant. In our study, patients with asthma are at higher risk of recurrence. PMID- 15126736 TI - Patient perception of comorbid conditions after acoustic neuroma management: survey results from the acoustic neuroma association. AB - OBJECTIVES/HYPOTHESIS: Based on survey results of the Acoustic Neuroma Association, the patient ratings of the most difficult aspects of acoustic neuroma management were reported and a review of the literature was made regarding comorbid conditions associated with acoustic neuroma treatment and their impact on patient quality of life. STUDY DESIGN: Cohort study of 1940 patients who were members of the Acoustic Neuroma Association. METHODS: A detailed questionnaire was mailed to 2372 members of the Acoustic Neuroma Association to identify preoperative and postoperative symptoms, complications, and long-term effects on physical and psychosocial function. For 1940 respondents (81.8%) who reported the "most difficult aspect of the AN [acoustic neuroma] experience," the responses were analyzed by tumor size, surgical approach, and patient age and sex. Statistical analysis was performed using SPSS software. RESULTS: Respondents reported that the most difficult aspect of the acoustic neuroma experience was hearing loss (25.8%), followed by facial weakness (17.9%), eye problems (10.8%), and headache (10.5%). In order of frequency, men reported hearing loss, balance problems, perioperative surgical experience, and eye and facial weakness, and women reported hearing loss, facial weakness, eye problems, and headache. Facial weakness was a morbidity more often reported for men and women who had large tumors, who were young, or who had undergone the retrosigmoid approach. Balance dysfunction was significant in patients older than 75 years of age. In patients with small tumors, headaches and balance problems were frequently reported. CONCLUSION: In the large cohort study of patients with acoustic neuroma, perceptions regarding the impact of treatment illustrated why it is incumbent on physicians to understand the sentiments of patients with acoustic neuroma when counseling them and recommending optimal management strategies. PMID- 15126737 TI - Hypopharyngeal perforation near-miss during transesophageal echocardiography. AB - OBJECTIVES/HYPOTHESIS: The traditional blind passage of a transesophageal echocardiography probe transorally through the hypopharynx is considered safe. Yet, severe hypopharyngeal complications during transesophageal echocardiography at several institutions led the authors to investigate whether traditional probe passage results in a greater incidence of hypopharyngeal injuries when compared with probe passage under direct visualization. STUDY DESIGN: Randomized, prospective clinical study. METHODS: In 159 consciously sedated adults referred for transesophageal echocardiography, the authors performed transesophageal echocardiography with concomitant transnasal videoendoscopic monitoring of the hypopharynx. Subjects were randomly assigned to receive traditional (blind) or experimental (optical) transesophageal echocardiography. The primary outcome measure was frequency of hypopharyngeal injuries (hypopharyngeal lacerations or hematomas), and the secondary outcome measure was number of hypopharyngeal contacts. RESULTS: No perforation occurred with either technique. However, hypopharyngeal lacerations or hematomas occurred in 19 of 80 (23.8%) patients with the traditional technique (11 superficial lacerations of pyriform sinus, 1 laceration of pharynx, 12 arytenoid hematomas, 2 vocal fold hematomas, and 1 pyriform hematoma) and in 1 of 79 patients (1.3%) with the optical technique (superficial pyriform laceration) (P =.001). All traumatized patients underwent flexible laryngoscopy, but none required additional intervention. Respectively, hypopharyngeal contacts were more frequent with the traditional than with the optical technique at the pyriform sinus (70.0% vs. 10.1% [P =.001]), arytenoid (55.0% vs. 3.8% [P =.001]), and vocal fold (15.0% vs. 3.86% [P =.016]). CONCLUSION: Optically guided trans-esophageal echocardiography results in significantly fewer hypopharyngeal injuries and fewer contacts than traditional, blind transesophageal echocardiography. The optically guided technique may result in decreased frequency of potentially significant complications and therefore in improved patient safety. PMID- 15126738 TI - Treatment of benign positional vertigo in the elderly: a randomized trial. AB - OBJECTIVES/HYPOTHESIS: The objective of the study was to evaluate the efficacy of three therapeutic strategies (Semont maneuver, flunarizine, and no treatment) in patients with benign paroxysmal positional vertigo. STUDY DESIGN: Randomized prospective trial. METHODS: One hundred fifty-six consecutive patients older than 60 years of age who were affected by benign paroxysmal positional vertigo of the posterior semicircular canal were enrolled. The diagnosis was made on the basis of the history of recurrent sudden crisis of vertigo and positional-induced typical nystagmus after Dix-Hallpike positioning maneuver. Patients were randomly allocated to receive Semont liberatory maneuver (intended as a statoconia detachment maneuver), flunarizine, or no treatment. A post-treatment negative Dix Hallpike test result was considered as a proof of vertigo resolution. RESULTS: Cure rates with Semont maneuver were significantly higher (94.2%) than those obtained with flunarizine (57.7%) and no treatment (36.4%) (P <.001). Within a 6 month follow-up, relapse rates were lower among patients treated with Semont maneuver (3.8%) than those obtained with flunarizine (5.8%) and no treatment (21.1%). All patients with resolution of symptoms and negative Dix-Hallpike test results showed a great improvement in daily activities and quality of life (P <.001). CONCLUSION: Semont liberatory maneuver is the most successful therapy for benign paroxysmal positional vertigo and improves patients' quality of life. Diagnostic and therapeutic maneuvers are easy to perform and should be part of the medical knowledge of every general practitioner and geriatrician. PMID- 15126739 TI - Chronological changes of hearing in pediatric patients with large vestibular aqueduct syndrome. AB - OBJECTIVES: To describe the chronologic changes of hearing in patients with large vestibular aqueduct syndrome (LVAS) and identify the prognostic factors. STUDY DESIGN: A retrospective chart-review study. METHODS: Twelve consecutive patients with LVAS were recruited at Taipei Veterans General Hospital between July 1986 and July 2000. The records of serial pure-tone audiogram and high-resolution computer tomography were collected. The chronologic figures of pure-tone average (PTA) were sketched. The investigated variables included sex, laterality and type of hearing loss (HL), size of vestibular aqueduct, and the chronologic changes and configurations of hearing. Data were analyzed statistically. RESULTS: The degree of HL in early childhood was from moderate to profound. The worst hearing could be estimated by the hearing level in early childhood. Sixteen of 24 ears were stable. Bilateral, chronologically stepwise-downhill hearing occurred in only one patient (1/12) during the follow-up period. Chronologically, high-tone hearings were worse than those of low-tone, but fluctuations of high-tone hearing were smaller than those of low-tone. The sizes of vestibular aqueduct were predictive of the density of major depression and its depth. CONCLUSIONS: LVAS, a congenital disease, is characterized by fluctuating sensorineural HL. Most hearing at PTA remained stable at least in one ear chronologically. The standard deviation of hearing at 500 Hz was the only prognostic factor for the progression of PTA. An enlarged vestibular aqueduct affects fluctuations of hearing, but the pathogenesis of HL still remains unclear and deserves further investigations. PMID- 15126740 TI - Atypical sinusitis in adults must lead to looking for cystic fibrosis and primary ciliary dyskinesia. AB - HYPOTHESES/OBJECTIVES:: In adults, purulent pansinusitis or nasal polyposis starting early in life or that is permanently infected or associated either with chronic bronchial infection, infertility, or situs inversus are uncommon. In these atypical cases of chronic sinusitis (ACS), a primary dysfunction of the mucociliary clearance can be suspected. Adult patients with ACS were therefore investigated to detect primary ciliary dyskinesia (PCD) or cystic fibrosis (CF). STUDY DESIGN: Open, prospective study. PATIENTS AND METHODS: Forty-two patients with ACS were investigated with ciliary beat frequency and ultrastructure analysis in nasal cells and cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation analysis in blood leukocytes. RESULTS: The diagnosis of PCD was confirmed in seven (17%) patients. At least one CFTR gene mutation was detected in 16 (38%) patients. The diagnosis of CF was suggested in three (7%) compound heterozygous patients. Another 13 (31%) patients were heterozygous for a CFTR gene mutation or a complex allele. Comparison of clinical features of ACS showed that only a family history of chronic sinusitis (P <.01) or chronic bronchitis (P <.02) and the presence of diffuse bronchiectasis (P <.0001) or serous otitis media (P <.0001) were significantly more frequent in PCD patients than in patients carrying CFTR gene mutations or those without PCD or CFTR gene mutations. CONCLUSIONS: ACS should be considered a remarkable entity in which congenital abnormalities of epithelial cells are frequently detected (55% of patients). The higher frequency of mutations in ACS patients compared with the general population suggests that heterozygoty for CFTR gene mutation could be a sinusitis-causing status. PMID- 15126741 TI - Laser myringotomy versus ventilation tubes in children with otitis media with effusion: a randomized trial. AB - OBJECTIVES: Insertion of ventilation tubes in children with otitis media with effusion (OME) is an accepted and common treatment procedure. The majority of patients require general anesthesia. Although laser myringotomy can be performed in local anesthesia, evidence is lacking that this treatment modality is an alternative for tubes, and outcome predictors for laser myringotomy are not available. STUDY DESIGN: Prospective randomized trial. METHODS: We screened 1,403 children with chronic OME that were indicated for placement of ventilation tubes. In the eligible patients, we performed laser myringotomy in one ear and placed a tube in the other ear, both within the same patient. Follow-up was scheduled each month for 6 months. Success was defined as absence of effusion or aural discharge. A logistic regression model was used with success of the therapy as binary outcome. This model was based on base-line variables, asked for in a parent's questionnaire. RESULTS: Two hundred eight children received the allocated intervention, and no complications occurred. The mean closure time of the laser perforation was 2.4 weeks, and the mean patency time of the ventilation tube was 4.0 months. The mean success rate was 40% for laser and 78% for tubes. Ten known variables were found to predict middle ear status after therapy. CONCLUSION: Laser myringotomy is a safe but less-effective procedure than insertion of a ventilation tube in the treatment of chronic OME. The prognostic model enables the otolaryngologist to choose the surgical treatment for the child that benefits most: laser myringotomy or ventilation tube. PMID- 15126743 TI - Epstein-Barr virus (EBV) latent membrane protein 1 induces interleukin-8 through the nuclear factor-kappa B signaling pathway in EBV-infected nasopharyngeal carcinoma cell line. AB - BACKGROUND/OBJECTIVES: Nasopharyngeal carcinoma (NPC) is a highly invasive and metastatic malignant tumor and is associated with Epstein-Barr virus (EBV) infection that exhibits type II latency. Angiogenesis is essential for tumor growth, invasion, and metastasis. Our previous studies have indicated that interleukin (IL)-8 was over-expressed in many NPC tissues and was found to be significantly correlated with angiogenesis by immunohistochemistry. STUDY DESIGN: In vitro design. METHODS: The influence of the EBV genome for IL-8 gene expression was studied using the EBV-genome-positive and -negative epithelial/NPC hybrid cell line NPC-KT. The EBV-positive and -negative clones were selected by polymerase chain reaction and in situ hybridization. RESULTS: EBV-positive clones expressed abundant IL-8 mRNA compared with EBV-negative clones. This result indicated that over-expression of IL-8 depended on the presence of EBV genomes in NPC-KT cells. Two encoded genes, latent membrane protein (LMP)1 and EBV-encoded small RNAs (EBERs), expressed in NPC were transfected in EBV-negative NPC-KT cells. LMP1 transactivated the IL-8 promoter, whereas EBERs did not. Moreover, the nuclear factor (NF)-kappa B binding site in the IL-8 promoter was essential for the response to LMP1, and the activator protein (AP)-1 binding site played only a partial role. CONCLUSIONS: LMP1 induces IL-8 mainly through the activation of NF-kappa B and partly through AP-1 in NPC model cell lines, NPC-KT, and this suggests that LMP1 plays an important role in the angiogenesis of NPC. PMID- 15126742 TI - Acute otitis externa: efficacy and tolerability of N-chlorotaurine, a novel endogenous antiseptic agent. AB - OBJECTIVE: The study's objective was to test the tolerability and efficacy of the endogenous antiseptic N-chlorotaurine (NCT) in comparison with a standard clinical treatment according to a phase IIb clinical trial protocol. STUDY DESIGN: The antimicrobial agent NCT was compared with the antibiotic component drops Otosporin (containing neomycin, polymyxin B, and hydrocortisone) for topical treatment of acute otitis externa in a randomized and rater-blinded clinical study. METHODS: Fifty patients suffering from acute otitis externa were divided into two groups according to a randomized list. The test group was treated with 1 mL of 1% aqueous NCT solution, the reference group with 1 mL of Otosporin. The substances were applied to the external ear canal at one daily session until the signs of infection disappeared. Efficacy and tolerability were evaluated daily by visual analogue scale and a six-step infection score. In addition, smears were analyzed to identify the causative pathogens. RESULTS: Both medications were equally well tolerated by the patients. The treatment was successful for all patients of the NCT group, whereas in one patient from the reference group, the infection did not disappear. The inflammation score improved more rapidly in the NCT group, which resulted in an earlier termination of the therapy. This difference became highly significant on days 4 to 7 (P <.01 each). Time needed for disappearance of inflammation (score 0) was 5.6 +/- 1.6 (mean +/- SD, range 3-9) days in the NCT group and 7.4 +/- 1.6 (range 4-10) days in the Otosporin group (P <.001). As expected, microbiologic cultures from ear swabs revealed Pseudomonas aeruginosa (58%) followed by Staphylococcus aureus (18%) as the main causative pathogens. CONCLUSIONS: NCT appears to be well tolerated and more effective than the therapy using antibiotic component drops. Because of its endogenous nature and its higher efficacy, NCT appears to be a good choice for topical treatment of acute otitis externa. PMID- 15126744 TI - Delayed vertigo after stapes surgery. AB - OBJECTIVES: Stapes surgery restores partial or total hearing in almost 95% of cases, and in case of failure, revision surgery may often resolve the problem. Delayed vertigo is commonly related to perilymphatic fistula. The aim of this study is to report experience gained in revision stapes surgery in cases of delayed vertigo. STUDY DESIGN: This is an intervention study, before-after trial; it includes follow-up between 12 and 84 months that was based on clinical history and audiometric evaluations. METHODS: The work was carried out in the otologic surgery referral center of Piemonte in outpatient surgery. Nine patients (4 males and 5 females, between 43 and 60 years of age) who presented with delayed vertigo after stapes surgery were retrospectively reviewed. All nine underwent clinical history evaluation, pure tone audiogram, investigation of the vestibular system with a bithermal binaural caloric test, and fistula test. Vestibular tests were performed with electronystagmography recording. In all nine subjects, functional middle ear exploration was carried out by way of a transmeatal approach using local anesthesia. The demonstration of a perilymphatic leak was positive in only three (33%) cases, but the oval window region was filled with fibrin glue in all nine cases. RESULTS: At follow-up, vertigo was resolved in all cases with revision surgery, even though perilymph leak was positive only in three cases. CONCLUSION: From the results obtained, we feel that exploration of the middle ear should be always carried out in cases of delayed vertigo after stapes surgery with suspected perilymphatic fistula. PMID- 15126745 TI - Adenoidectomy: selection criteria for surgical cases of otitis media. AB - OBJECTIVE: Nasopharyngeal adenoids may serve as a mechanical obstruction to the eustachian tube and contribute to the pathophysiology of otitis media (OM). The purpose of this study was to determine whether abutment of adenoids laterally against the torus tubaris affects the outcome of patients requiring pressure equalization tubes (PET) for OM. STUDY DESIGN: Randomized, controlled, prospective clinical trial. METHOD: Patients requiring PET for recurrent acute OM or OM with persistent effusion were randomized into two groups: 1) PET placement and 2) PET placement and adenoidectomy, regardless of whether the adenoids were abutting or not abutting the torus tubaris. Patients were followed for a minimum of 1 year to determine rate of treatment failure, defined as recurrence of acute OM (>3 times/year), OM with effusion, or reinsertion of PET. RESULTS: Of the 34 patients in the abutting group, 16 patients underwent only PET insertion, of whom 8 (50%) failed, whereas 18 patients had combined PET placement and adenoidectomy, of whom 3 (17%) failed. There was a statistical difference between these two groups (P < 05). Of the 29 patients in the nonabutting group, 24 patients underwent only PET insertion, of whom 9 (37.5%) failed, whereas 5 patients underwent combined PET placement and adenoidectomy, of whom 2 (40%) failed. There was no statistical difference between these two groups (P =.92). CONCLUSION: This study demonstrates that the position of hypertrophied adenoids may alter the final otologic outcome of patients requiring PET insertion for OM. Patients with adenoids abutting the torus tubaris may benefit most from an adjuvant adenoidectomy. PMID- 15126746 TI - Tissue engineering of a human sized and shaped auricle using a mold. AB - OBJECTIVES: The creation of a tissue-engineered auricle was initially successful in an immunocompromised nude mouse model. Subsequently, an immunocompetent porcine model successfully generated a helical construct. We wished to evaluate the novel technique of using a mold to create a complete, anatomically refined auricle in a large animal model. METHODS: Mixtures of autogenous chondrocytes and biodegradable polymers were used inside a perforated, auricle shaped hollow gold mold. Three biodegradable polymers (calcium alginate, pluronic F-127, and polyglycolic acid) were used to retain the seeded chondrocytes inside the mold. These molds, along with a control, were implanted subcutaneously in the abdominal area of 10 animals (pigs and sheep). The constructs were removed after 8 to 20 weeks and were assessed by gross morphology and histology. RESULTS: All the gold implants were well tolerated by the animals. The implants using calcium alginate (n = 3) generated constructs of the exact shape and size of a normal human ear; the histology demonstrated mostly normal cartilage with some persistent alginate. The implants with pluronic F-127 (n = 3) resulted in cartilage with essentially normal histology, although leakage outside the molds and external cartilage generation was noted. Polyglycolic acid implants (n = 3) produced no useful cartilage because of an inflammatory reaction with fibrosis. The empty control mold (n = 1) demonstrated only a very small amount of fibrous tissue inside. CONCLUSION: A tissue-engineered human sized auricle of normal anatomic definition can be generated in an immunocompetent large-animal model using a mold technique. Although further refinements will be necessary, the technique appears promising for potential use in patients with microtia. PMID- 15126747 TI - Pediatric tonsillotomy with the radiofrequency technique: less morbidity and pain. AB - OBJECTIVE: To compare two techniques for pediatric tonsil surgery with respect to pain and postoperative morbidity. The two methods were the partial tonsil resection using radiofrequency (RF) technique (tonsillotomy [TT]) versus traditional tonsillectomy (TE). STUDY DESIGN: Prospective clinical randomized study in one tertiary care ENT clinic and two secondary care clinics. METHOD: One hundred fifty children, between 5 and 15 years of age, were randomized to either TT with RF using the Surgitron Ellman, 1.7 MHz, or regular TE. Randomization was performed from the waiting list, including children with both a history of obstructive problems and recurrent tonsillitis. The TT was performed with a specially made sling electrode using a cut/coagulation mode. RESULTS: Forty-nine children were operated on with TT and 43 with TE. There was significantly less bleeding in the TT group, although two cases of primary postoperative bleeding occurred among the TT children and one in the TE group. The pain recordings showed significantly less pain for the TT children from the second hour postoperatively onward, and the TT children were pain free and in school 3 days earlier than the TE group. The TT group had less need of the prescribed drugs (diclofenac and paracetamol). After 9 days, 73% of the TT children were completely healed, but only 31% of the TE children. By that time, the TE children had lost a mean of 660 g, and the TT children had gained 127 g. The effect on snoring was the same for both groups. CONCLUSION: RF appears to be a safe and reliable method for tonsil surgery with much less postoperative morbidity than regular TE. PMID- 15126748 TI - A laryngeal dissection station: educational paradigms in phonosurgery. AB - OBJECTIVES: To introduce a new tool for phonosurgical training and education. A multitude of innovations in complex laryngeal surgery has catalyzed new educational initiatives. Establishing dexterity in phonomicrosurgery is often difficult to achieve while working on patients because of the narrow margin for success. Furthermore, laryngoplastic phonosurgery and open partial laryngectomy require sophisticated knowledge of precise anatomic relationships, which can be difficult to express in images. Finally, many teaching programs do not have a high volume of these procedures, and there is a significant need to transmit this information in continuing education courses. STUDY DESIGN: Prototype design. METHODS: A laryngeal dissection station (LDS) was designed to facilitate the acquisition of high-level procedural skill sets for both transoral and transcervical techniques. RESULTS: This LDS can be used in existing temporal-bone laboratories by using cadaveric larynges. A rectangular frame supports two adjustable holders, one for the larynx and one for the examining speculum of a laryngoscope. Procedures are performed with the larynx fixed in space by a novel fixator. Variation in position and orientation of the components affords simulation of both microlaryngoscopy and open surgery. The dissection station can accommodate virtually any laryngoscope, regardless of size or shape. CONCLUSIONS: This training apparatus should facilitate laryngeal surgical instruction in residency training and continuing medical education. This device and others like it can help establish clinical competency in laryngology, should this become necessary in future educational models of residency training and recertification. PMID- 15126749 TI - Middle ear pressure changes after nitrous oxide anesthesia and its effect on postoperative nausea and vomiting. AB - OBJECTIVES/HYPOTHESIS: This study was designed to explore the relationship between changes in middle ear pressure associated with inhalational anesthesia and the incidence of postoperative nausea and vomiting (PONV). STUDY DESIGN: Prospective, randomized. METHODS: Middle ear compartment pressures were measured by tympanometry in 27 randomly assigned knee arthroscopy patients throughout the surgical procedure as well as into recovery. RESULTS: A positive correlation between the maximum positive pressure (MPP) and maximum negative pressure (MNP) gradient and PONV was demonstrated (P <.05). The incidence of PONV in the nitrous oxide (N2O) treatment group was 6 of 16 patients, whereas only 2 of 11 patients in the control group developed nausea, vomiting, and vertigo symptoms. Those patients that did not experience PONV demonstrated a median MPP of 155 with a median MNP of -52. The patients that experienced PONV exhibited a median MPP of 179 with a median MNP of -164. This demonstrates a significant increase in the incidence of PONV in the N2O treatment group. CONCLUSIONS: Barometric changes in the middle ear contribute to the incidence of PONV induced by N2O. PMID- 15126750 TI - Microbial superantigens induce glucocorticoid receptor beta and steroid resistance in a nasal explant model. AB - OBJECTIVE: To study the role of superantigen (SAg) in inducing glucocorticoid (GC) receptor beta and steroid resistance in an explant model of nasal tissue. METHODS: Nasal tissue was obtained from inferior turbinates of controls and ragweed (RW)-sensitive patients. Tissue samples were incubated with SAg of staphylococcal enterotoxin B. In addition, tissue samples from RW-sensitive patients were incubated with RW allergen in the presence and absence of both SAg and dexamethasone (DEX). The expression of GC receptor beta was assessed by immunocytochemistry. The expression of interleukin (IL)-2 and IL-4 mRNA was assessed by in situ hybridization. RESULTS: SAg induced an increase in the expression of GC receptor beta in atopic tissue and to a lesser extent in nonatopic tissue. The most significant induction of GC receptor beta was observed in response to SAg and RW in atopic tissue. Stimulation of atopic tissue with RW alone and SAg alone induced IL-4 and IL-2 mRNA, respectively. Incubation of atopic tissue with both SAg and RW induced both IL-2 and IL-4 mRNA. The increase in IL-4 mRNA expression was blunted by the addition of DEX to atopic tissue stimulated with RW alone but not to tissue stimulated by both RW and SAg. CONCLUSION: Our results demonstrate that SAgs induce steroid resistance in atopic nasal explant tissue by up-regulating the expression of GC receptor beta. Furthermore, we have shown that the up-regulation of GC receptor beta is a local event that is associated with the coexpression of IL-2 and IL-4 mRNA. PMID- 15126751 TI - Maxillomandibular expansion for the treatment of sleep-disordered breathing: preliminary result. AB - OBJECTIVE: To assess the outcomes of maxillomandibular expansion (MME) by distraction osteogenesis (DO) for the treatment of sleep-disordered breathing (SDB). METHODS: This was a prospective study of six consecutive patients with SDB. All of the patients have maxillary and mandibular constriction and were treated with MME. Variables examined include age, sex, body mass index (BMI), polysomnographic results (PSG), Epworth Sleepiness Scale (ESS), and the extent of the widening of the maxilla and mandible. RESULTS: All six patients (4 males) completed MME for the treatment of SDB. The mean age was 22.2 +/- 11.4 years. The mean maxillary expansion was 10.3 +/- 3.0 mm, and the mean mandibular expansion was 9.5 +/- 2.9 mm. ESS improved from 10.2 +/- 1.9 to 5 +/- 2.9. The mean apnea/ hypopnea index (AHI) improved from 13.2 +/- 15.6 to 4.5 +/- 5.8 events per hour, and the mean lowest oxygen saturation (LSAT) improved from 88.2 +/- 2.9% to 91.3 +/- 3.3%. The mean esophageal pressure improved from -20 +/- 11.3 cm H2O to -8 +/ 3.6 cm H2O. No complications were encountered, and the follow-up period was 18.1 +/- 9.8 months. CONCLUSION: : The result suggests that MME improves SDB in patients with maxillary and mandibular constriction and can be a valid treatment. PMID- 15126752 TI - Adaptation of the horizontal vestibuloocular reflex in pilots. AB - OBJECTIVES/HYPOTHESIS: The vestibular function in pilots has been reported as being different to that in other, normal subjects. However, there is little evidence as to why and when this occurs, and its cause is still in doubt. The aim of this study was to evaluate changes in vestibular function in student pilots attending an elementary flight course, and in other active pilots, through the use of the slow harmonic acceleration (SHA) test and to verify the occurrence and timing of any vestibular function changes. STUDY DESIGN: Cross-sectional and prospective group design. METHODS: The six study groups were control, student pilots before and after flight training, and three groups of active pilots classified according to flight time. Gain, phase, and symmetry of horizontal eye movements induced by sinusoidal harmonic acceleration test were investigated. RESULTS: There were significantly higher gain values in all active pilot groups as compared with the control group at frequencies above 0.04 Hz, and in student pilots after flight training, as compared with before training at 0.01, 0.04, and 0.08 Hz. However, there were no significant differences in gain values among the three groups of active pilots or between gain values in student pilots before flight training and those of the control group. We could not find any reliable relationship between vestibuloocular reflex (VOR) function changes and other factors such as age, height, and weight. CONCLUSIONS: We suggest that these results might be explained by adaptations caused by VOR plasticity rather than habituation and that flight training might be responsible for the modulation of the vestibular function in pilots. PMID- 15126753 TI - Regression of a sphenochoanal polyp in a child. AB - OBJECTIVES: To present an unusual case of a sphenochoanal polyp that regressed and review the etiology of such polyps in comparison to the commoner antrochoanal polyp. STUDY DESIGN: Case study. METHODS: One was incidentally discovered in a girl aged 3 years, 8 months at postnasal mirror examination after adenotonsillectomy. RESULTS: Computed tomography scans 1 month later confirmed the polyp, but three months later, MRI scans only revealed sphenoid sinus opacification. Sphenoidotomy revealed normal sinuses implying inflammation had resolved. CONCLUSION: This patient is believed to be the youngest reported to have sphenochoanal polyp, and the only one where the polyp had completely regressed. This regression suggests that sphenochoanal polyps may be more common than is thought but are mostly asymptomatic. PMID- 15126754 TI - Merkel cell carcinoma. AB - PURPOSE: To determine the natural history and treatment outcomes for patients with Merkel cell carcinoma. METHODS: Review of the literature. RESULTS: The probability of regional node involvement at presentation exceeds 50%; few patients present with distant metastases. Comprehensive treatment of the primary site and regional lymphatics with surgery or radiotherapy results in the highest likelihood of cure. The role of adjuvant chemotherapy remains investigational. CONCLUSION: The probability of regional dissemination at diagnosis is high. The optimal treatment is resection of the primary tumor and treatment of the regional lymphatics. Resection of all gross tumor should be accomplished followed by local regional radiotherapy in most patients. PMID- 15126755 TI - The unfinished Turandot and Puccini's laryngeal cancer. PMID- 15126756 TI - Squamous cell carcinoma of the oral tongue in young patients. AB - OBJECTIVES/HYPOTHESIS: Cancer of the tongue is reported with increasing frequency in young people. The objective of this work was to study the biologic and clinical course of the disease in this group. The clinical course of the disease in this patient group remains controversial. STUDY DESIGN: A retrospective 30 year review was made from data from a tertiary academic medical center. METHODS: A chart review was performed for 48 patients with oral tongue cancer. The following variables were compared: age, sex, tobacco and alcohol use, treatment, and outcome. RESULTS: Stage (T1,2-N0) and treatment modality were similar in the two age groups, as was disease-specific outcome. However, in the younger group, the clinical course followed two distinct patterns: extremely aggressive appearance with a 40% mortality rate within 2 years compared with 10.7% in the older group (P <.05) or indolent with freedom from disease for over 15 years. CONCLUSIONS: Squamous cell carcinoma (SCC) of the oral tongue may be associated with a variable clinical course in younger patients. Although, in general, the disease-specific outcome is similar to that of the older patients, some of the patients have an exceptionally aggressive disease. An extensive therapeutic regimen should be used in all cases. PMID- 15126757 TI - Human cortical motor representation of the larynx as assessed by transcranial magnetic stimulation (TMS). AB - OBJECTIVES: To analyze characteristic features and details on motor-evoked potentials (MEPs) of the cricothyroid and vocalis muscles from single-pulse cortical transcranial magnetic stimulation (TMS) in normal subjects to characterize cortical motor representation of laryngeal muscles. STUDY DESIGN: Prospective, experimental investigation on healthy volunteers. METHOD: MEPs of the cricothyroid and vocalis muscles elicited by cortical TMS with a figure-8 shaped coil were investigated in two groups of six healthy subjects each, with special regard to MEP amplitude as a function of the coil position on the head surface along the interaural line. RESULTS: Bilateral reproducible responses of the cricothyroid and the vocalis muscles could be observed in all subjects. For the cricothyroid muscle, maximal responses were obtained at mean stimulus positions of 7.5 +/- 1.4 cm (contralateral) and of 7.3 +/- 1.3 cm (ipsilateral), respectively. For the vocalis muscle, we found maximal responses at mean stimulus positions of 10.3 +/- 1.9 cm (contralateral) and of 9.6 +/- 1.6 cm (ipsilateral), respectively. Despite a considerable overlap of these coil positions, from which reproducible MEPs could be elicited in both groups of the laryngeal muscles, statistically significant separation of the cricothyroid-and vocalis-associated cortical representation areas was possible. CONCLUSIONS: Our observations point to two different cortical motor representation areas, with the cricothyroid muscle-related area being located more medially. PMID- 15126758 TI - Evaluation of the medical and surgical treatment of chronic rhinosinusitis: a prospective, randomised, controlled trial. AB - OBJECTIVES: To conduct the first prospective, randomized, controlled trial evaluating and comparing the medical and surgical treatment of polypoid and nonpolypoid chronic rhinosinusitis (CRS). MATERIALS AND METHODS: Ninety patients with CRS were equally randomized either to medical or surgical therapy. All patients underwent pre- and posttreatment assessments of visual analogue score (VAS), the Sinonasal Outcome Test-20 (SNOT-20), the Short Form 36 Health Survey (SF-36), nitric oxide (NO), acoustic rhinometry, saccharine clearance time (SCT), and nasal endoscopy. Each patient had three assessments: before starting the treatment, after 6 months, and, finally, after 1 year. RESULTS: Both the medical and surgical treatment of CRS significantly improved almost all the subjective and objective parameters of CRS (P <.01), with no significant difference being found between the medical and surgical groups (P >.05), except for the total nasal volume in CRS (P <.01) and CRS without polyposis (P <.01) groups, in which the surgical treatment demonstrated greater changes. CONCLUSION: CRS should be initially targeted with maximal medical therapy (e.g., a 3 month course of a macrolide antibiotic, douche, and topical steroid), with surgical treatment being reserved for cases refractory to medical therapy. The presence of nasal polyps is not a poor prognostic factor for the efficacy of CRS therapy, either surgical or medical. PMID- 15126759 TI - Effects of external beam radiation in the rat tibial nerve after crush, transection and repair, or nerve isograft paradigms. AB - INTRODUCTION: In head and neck surgery, radiation therapy is often administered to an injured nerve. Previous studies have examined the effects of either preoperative or postoperative radiation on nerve regeneration in rodents. In these studies, histomorphometric analysis was performed up to 8 month postoperatively. Given the exceptional neuroregenerative capacity of rodents, significant differences in nerve regeneration may go undetected if nerves are evaluated at such distant postoperative time points. This study is designed with a more appropriate model and investigates the effects of radiation after three common nerve injury paradigms. METHODS: Sixty-four Lewis rates were randomized to 8 groups corresponding to uninjured, tibial nerve crush, transection and repair, or reconstruction with isografts. Half of the animals in each of these paradigms (n = 8 per group) were treated with 10 Gy of external beam radiation to the site of nerve injury at 7 days postoperatively. On postoperative day 28, functional recovery and histomorphometric assessment was performed. RESULTS: For a given paradigm of nerve injury, no significant differences in nerve fiber number, neural density, neural debris, or fiber width were noted between the control and radiated groups, and radiation did not affect functional recovery. CONCLUSION: Radiation had no discernible effect on nerve regeneration or functional recovery in the rodent nerve injury models studied. All assessments were made at time points suitable for detecting differences in nerve regeneration between groups. These findings suggest that administration of radiation to fields containing injured peripheral nerve is unlikely to adversely affect functional outcomes. PMID- 15126760 TI - Incidence of methicillin-resistant Staphylococcus aureus causing chronic rhinosinusitis. AB - OBJECTIVE/HYPOTHESIS: To identify the incidence of methicillin-resistant Staphylococcus aureus (MRSA) causing chronic rhinosinusitis (CRS) and identify whether antibiotic use and previous endoscopic sinus surgeries (ESS) contribute to its development. STUDY DESIGN: A retrospective case control analysis, with the control group randomly selected and matched for age and sex. METHODS: All patients undergoing an endonasal culture in a tertiary otolaryngology center between April 2001 and March 2003 for mucopurulent rhinosinusitis were identified. A chart review was undertaken to identify those patients with a positive MRSA culture result. An age- and sex-matched control group was randomly chosen, and an interview was conducted to identify antibiotic use and previous ESS. A statistical analysis on these two variables was carried out using a t test and the Wilcoxon rank-sum test. RESULTS: Two hundred eighty cultures were identified in 188 patients, and of these, 264 cultures in 173 patients met the inclusion criteria. There were 141 positive cultures and 13 positive MRSA cultures. The overall incidence of MRSA in this population was 9.22%. The mean number of antibiotic courses in the MRSA and the control groups was 33.2 and 26.7, respectively, which was not statistically significant (P =.43). The 95% confidence interval (CI) for this mean difference of 6.5 is -9.8 to 22.8. The mean number of ESS in the MRSA and control groups was 2.0 and 1.9, respectively, which was not statistically significant (P =.93). The 95% CI for this mean difference of 0.06 is -1.3 to 1.4. CONCLUSIONS: We identified a 9.22% incidence of MRSA-causing CRS. The frequency of antibiotic use and previous ESS were found not to be statistically significant causes of MRSA sinusitis. However, the 95% CI for antibiotic usage is skewed to the right, indicating a possible role for its contribution to the emergence of MRSA-causing CRS. PMID- 15126761 TI - Bone-anchored hearing aid: comparison of benefit by patient subgroups. AB - OBJECTIVES/HYPOTHESIS: The osseointegrated bone-anchored hearing aid, using the Branemark system, is well established and has proven benefit. The aim was to study quality of life benefits within patient subgroups using the validated Glasgow Benefit Inventory (GBI). STUDY DESIGN: Retrospective questionnaire study. METHODS: Ninety-four consecutive patients were enrolled into the study. Mean patient age was 49 years, with a female-to-male ratio of 1.1:1. Patient subgroups were discharging mastoid cavities, chronic active otitis media, congenital ear problems, otosclerosis, and acoustic neuroma and other unilateral hearing losses. RESULTS: The response rate was 73%. The score for total benefit of bone-anchored hearing aid fitting for the entire group was +33.3 (95% confidence interval [CI], 25-42). Glasgow Benefit Inventory scores for each subgroup were all greater than +20. The congenital atresia group scored highest with +45 (95% CI, 28-61). Variation in benefit across the subgroups has been demonstrated. Fitting of BAHA following acoustic neuroma surgery was shown to be of benefit with a score of +22.2. General benefits scored highest in all subgroups compared with physical and social benefits. CONCLUSION: The study demonstrated the differences in benefit within patient subgroups. Its results can be used to give patients a predictive value at the time of preoperative counseling. The study identified congenital ear disorders as the group likely to obtain maximal benefit. Notably, for the first time, the study demonstrated the documented benefit of restoring stereo hearing to patients who have acquired unilateral hearing loss following acoustic neuroma surgery using a BAHA. PMID- 15126762 TI - Frontal sinus complications after frontal craniotomy. AB - OBJECTIVES: To review frontal sinus complications following frontal craniotomy and to describe management strategies. STUDY DESIGN: Retrospective review. METHODS: Retrospective review was made of six patients who had undergone frontal craniotomy and subsequently developed frontal sinus complications. Demographic data, indication for craniotomy, type of reconstruction, average time to development of complications, presenting symptoms, diagnosis, surgical management, follow-up, and outcomes were reported. RESULTS: Complications included unilateral frontal sinus mucoceles in four patients, bilateral frontal sinus mucoceles in one patient, and bilateral frontal sinus mucopyoceles with upper-eyelid abscess in one patient. The average time to presentation of symptoms and development of complications following frontal craniotomy was 14.8 years (range, 1-39 y). Headaches were the most common presenting complaint. All patients underwent endoscopic mucocele marsupialization as part of their management. After an average follow-up period of 9 months, no recurrences were found and no complications occurred. CONCLUSION: A small number of patients develop otolaryngological complications, most commonly, frontal mucoceles, following frontal craniotomy. A high level of suspicion and long-term surveillance are needed to monitor for their occurrence. Endoscopic marsupialization may provide an effective, safe means for management. PMID- 15126763 TI - Primary parotid malignoma surgery in patients with normal preoperative facial nerve function: outcome and long-term postoperative facial nerve function. AB - OBJECTIVES/HYPOTHESIS: Most patients with primary parotid cancer present with normal facial nerve function. The common surgical strategy for these patients is to perform a parotidectomy with facial nerve preservation. Nevertheless, the functional outcome for the facial nerve and oncological outcome is unclear. STUDY DESIGN: Medical records of 211 patients treated from 1986 to 2000 in an university hospital were reviewed. One hundred seven patients with primary surgery for primary parotid cancer and long-term follow-up were analyzed retrospectively. METHODS: The characteristics, treatment, and oncological outcome were evaluated using hospital chart data. All patients with postoperative facial paresis were had follow-up with electromyography until recovery or permanent paresis became apparent. RESULTS: Ninety-one patients had a normal preoperative function. Facial nerve preservation during surgery by total parotidectomy was possible in 79 patients, whereas in 28 patients a radical parotidectomy was necessary. Otherwise, the oncological characteristics of both groups were not different. Directly after total parotidectomy, half of the patients presented a facial paresis but only two patients (2%) developed a permanent partial paresis. The 5-year disease-free rate and the 5- and 10-year survival rates were 65%, 83%, and 54%, respectively. After radical parotidectomy, the results were not significantly different. The 5-year disease-free rate and the overall 5- and the 10-year survival rates were 56%, 62%, and 42%, respectively. CONCLUSION: Treatment of primary parotid cancer with preoperatively normal facial nerve function by standardized parotidectomy and precise microsurgical preservation of the facial nerve is often possible. This approach demonstrates favorable oncological results with a low level of long-term facial nerve morbidity. PMID- 15126764 TI - Use of soft tissue vascular flaps for mastoid cavity obliteration. PMID- 15126769 TI - Practice and progress in kidney cancer: methodology for novel drug development. AB - PURPOSE: The minimal efficacy of standard therapy for metastatic renal cell carcinoma (RCC) has resulted in the evaluation of numerous novel agents. Some agents have shown promise in phase II trials and yet none have improved survival over standard therapy in phase III trials. We examined existing data relevant to standard therapy and clinical trial methodology in RCC to understand patient, disease and trial design factors that have impacted clinical trial outcome and drug evaluation. Furthermore, we describe new paradigms for the evaluation of novel agents to optimize the yield of clinical research in RCC. MATERIALS AND METHODS: A comprehensive review of published retrospective analyses and phase II/phase III trials in patients with metastatic RCC was undertaken. Publications with patient selection and/or therapeutic implications in our judgment are presented and evaluated. RESULTS: Patients with good performance status and access to centers with experienced staff may appropriately receive high dose interleukin-2 after consideration of the relative risks and benefits. Alternatively low dose, single agent cytokine regimens are acceptable. Novel agents may be tested in untreated and refractory RCC. Consideration of the prognostic factors of a given phase II cohort is essential when interpreting single arm clinical trial results. RCC histological subtypes continue to be distinguished biologically and treatment relevant to the vascular endothelial growth factor pathway is most appropriately targeted to clear cell RCC. Nephrectomy in metastatic RCC may impact evaluation of the tumor response and survival in metastatic RCC. Thus, consideration of nephrectomy status in phase II trials and stratification in phase III trials is warranted. Clinical trials that include patients with central nervous system metastases should have standardized treatment of these metastases prior to systemic therapy. Objective response rate as an end point should be used with caution, given its unreliable history in metastatic RCC. Novel trial designs using time to disease progression may allow for interpretation of the antitumor effect in the absence of tumor shrinkage. CONCLUSIONS: Metastatic renal carcinoma is a model disease for the design of clinical trials and testing of novel agents. Novel trial designs and end points should be considered to evaluate new agents in RCC. Phase III trials must be carefully performed with the most promising agents to impact survival in this disease. PMID- 15126770 TI - Baseline staging of newly diagnosed prostate cancer: a summary of the literature. AB - PURPOSE: Staging for prostate cancer often includes bone scanning and computerized tomography (CT). We systematically reviewed the published evidence for these tests. MATERIALS AND METHODS: We searched MEDLINE for articles on these investigations in newly diagnosed cases of prostate cancer. Data were pooled based on prostate specific antigen (PSA), grade and tumor stage. RESULTS: Among 23 studies examining the role of bone scan metastases were detected in 2.3%, 5.3% and 16.2% of patients with PSA levels less than 10, 10.1 to 19.9 and 20 to 49.9 ng/ml, respectively. Scanning detected metastases in 6.4% of men with organ confined cancer and 49.5% with locally advanced disease. Detection rates were 5.6% and 29.9% for Gleason scores 7 or less and 8 or greater, respectively. Among 25 studies CT documented lymphadenopathy in 0 and 1.1% of patients with PSA less than 20 and 20 ng/ml or greater, respectively. CT detection rate was 0.7% and 19.6% in patients with localized and locally advanced disease, respectively. Detection rates in patients with Gleason scores 7 or less and 8 or greater were 1.2% and 12.5%, respectively. These risks were typically much greater on pathological evaluation. CONCLUSIONS: Patients with low risk prostate cancer are unlikely to have metastatic disease documented by bone scan or CT. Therefore, these investigations should not be standard practice. However, patients with PSA 20 ng/ml or greater, locally advanced disease, or Gleason score 8 or greater are at higher risk for bone metastases and should be considered for bone scan. CT may be useful in patients with locally advanced disease or Gleason score 8 or greater but appears not to be of benefit in patients with increased PSA alone. PMID- 15126771 TI - Emerging role of botulinum toxin in the management of voiding dysfunction. AB - PURPOSE: In recent years there has been tremendous excitement over the use of botulinum neurotoxin (BTX) to treat various urethral and bladder dysfunctions. BTX is the most potent, naturally occurring toxin known to mankind. Why, then, would a urologist want to use this agent to poison the bladder or urethral sphincter? MATERIALS AND METHODS: We reviewed the recent literature on the mechanisms underlying the effects of BTX treatment and discuss current use of this agent within the urological community, as well as provide perspective on future targets of BTX. The information was gathered from MEDLINE, abstracts from recent urological meetings and personal experience. RESULTS: Injection of BTX appears to have a positive therapeutic effect in multiple urological conditions, including detrusor hyperreflexia and detrusor external sphincter dyssynergia, and nonneurogenic conditions such as pelvic floor spasticity, refractory overactive bladder and, possibly, benign prostatic hyperplasia. Interstitial cystitis may even be potentially helped with bladder BTX injection. CONCLUSIONS: Botulinum toxin is a novel and promising treatment for a variety of lower urinary tract dysfunctions. The basic science behind its mechanism of action and physiology, and published clinical results are impressive. However, since application of BTX in the lower urinary tract has not been approved by the Food and Drug Administration, caution should be used until future properly designed, multicenter randomized studies are completed to assess the safety and efficacy of BTX in urological diseases. PMID- 15126772 TI - Long-term outcome of patients with interstitial cystitis treated with low dose cyclosporine A. AB - PURPOSE: We evaluated patients with interstitial cystitis who had been on cyclosporine A treatment for at least a year. Symptom improvement on micturition charts and subjective expression of bladder pain were recorded. Side effects and safety of medication were evaluated. MATERIALS AND METHODS: A total of 23 patients (20 females and 3 males) fulfilling National Institute for Diabetes and Digestive and Kidney Diseases criteria of interstitial cystitis were included in this study. Age of patients at followup was 65.7 +/- 7.6 years (mean +/- SD). Mean followup was 60.8 +/- 35.7 months. Before starting cyclosporine A treatment multiple first line therapies had been tried without clinical help. RESULTS: The number of voidings in 24 hours was 20.8 +/- 6.3 before treatment. After a year of cyclosporine A treatment it was decreased to 10.2 +/- 3.8 (p < 0.001). Maximal bladder capacity increased from 161.8 +/- 74.6 to 360.7 +/- 99.3 ml in a year (p < 0.001). Mean voided volume increased from 101.4 +/- 42.7 to 246.4 +/- 97.9 ml (p < 0.001). The effect was maintained throughout followup. Of 23 total patients 20 reported no bladder pain on cyclosporine A treatment and 11 patients stopped treatment due to a good clinical effect. In 9 patients symptoms recurred within months but disappeared again after cyclosporine A treatment was restarted. Side effects of medication were infrequent. CONCLUSIONS: Cyclosporine A treatment was safe and effective in treating interstitial cystitis. The achieved therapeutic effect was maintained in the long term. Cessation of medication led to recurrence of symptoms in most cases. PMID- 15126773 TI - Midstream urine culture and sensitivity test is a poor predictor of infected urine proximal to the obstructing ureteral stone or infected stones: a prospective clinical study. AB - PURPOSE: This prospective study was performed to determine if midstream urine (MSU) culture and sensitivity (C&S) alone could adequately predict infected urine proximal to the obstructing ureteral stone or the infected stones. Can pelvic urine C&S predict infected stones? MATERIALS AND METHODS: A prospective clinical study was performed on all patients undergoing ureterorenoscopy and lithotripsy for ureteral stones with obstruction between December 1, 2000 and January 31, 2002. We obtained MSU, renal pelvic urine and fragmented stones for culture and sensitivity. An analysis of the data was performed to assess statistical association. RESULTS: A total of 73 patients who fulfilled the criteria were recruited. Of these patients 25 (34.3%) had positive stone culture, 43 (58.9%) had positive pelvic urine and 21 (28.8%) patients had positive MSU C&S. Stone and pelvic C&S were positive simultaneously in 17 (23.3%) cases, MSU and stone C&S were positive in 8 (10.9%) cases, whereas pelvic and MSU C&S were positive in 13 (16.4%) cases (p = 0.03). MSU C&S had a sensitivity of 30.2% and specificity of 73% to detect pelvic urine C&S positivity. MSU C&S had a low positive predictive value and negative predictive value (NPV) in relation to infected pelvic urine (positive predictive value = 0.62, NPV = 0.42). Pelvic urine C&S had a NPV of 0.73 in detecting noninfected stones. CONCLUSIONS: The results of this study suggest that in obstructive uropathy secondary to a stone MSU C&S is a poor predictor of infected urine proximal to the obstruction and infected stones. PMID- 15126774 TI - Comparison of guide wires in urology. Which, when and why? AB - PURPOSE: We compared and contrasted various guide wires with regard to their physical properties as they apply to their use for the access or coaxial passage of other catheters. MATERIALS AND METHODS: Certain 0.035-inch diameter guide wires were tested with regard to tip bending force, shaft bending force, pull force and tip puncture force, namely the Roadrunner PC and polytetrafluoroethylene (PTFE) wire guide (Cook Urological, Spencer, Indiana), Glidewire, Bentson type 15 cm flexible tip PTFE coated guide wire and Amplatz super stiff Urowire XF (Boston Scientific Microvasive, Miami, Florida), Bentson guide wire and Amplatz guide wire (Applied Medical, Rancho Santa Margarita, California) and the PTFE coated Bard guide wire (Bard Urological Division, Covington, Georgia). RESULTS: Regarding guide wires used for access, the Boston Scientific PTFE guide wire with a 15 cm flexible tip required the least amount of force to deflect the tip. Of the 3 cm flexible tip guide wires the Applied Bentson guide wire had the most flexible tip and the Bard guide wire had the stiffest flexible tip. The Boston Scientific Glidewire required the least amount of force to pull from a tortuous pathway and this guide wire also required the greatest force (4 times as much force as the other guide wires) to puncture the aluminum foil (p < 0.001), indicating the safety of its tip. Regarding axial rigidity for the coaxial passage of other catheters over a guide wire, the Boston Scientific Amplatz super stiff guide wire was significantly more resistant to bending than all of the other guide wires that we tested (p < 0.05). CONCLUSIONS: Brand name guide wires designed for the same purpose appear to differ markedly with regard to flexibility, lubricity and shaft stiffness. In general, floppy tip and nitinol based guide wires appear to be best used for access with an emphasis on tip flexibility and a low friction coating, while the stiffer shaft guide wires are selected for coaxial passage of catheters, stents and sheaths. PMID- 15126775 TI - Tumor seeding in urological laparoscopy: an international survey. AB - PURPOSE: During the last 10 years laparoscopy has been applied to treat most urological pathology including malignancies. There has been concern regarding peritoneal dissemination and port site metastases. We undertook a survey to assess the incidence of this occurrence. MATERIALS AND METHODS: A total of 50 international urology departments with experts in laparoscopic urological surgery were contacted for this study. Each site was asked to complete a 2-page survey regarding the volume of laparoscopic urological procedures and port site recurrences. RESULTS: Nineteen sites elected to participate. A total of 18750 laparoscopic procedures were performed, of which 10912 were for cancer. These included 2604 radical nephrectomies, 559 nephroureterectomies, 555 partial nephrectomies, 27 segmental ureterectomies, 3665 radical prostatectomies, 1869 pelvic lymph node dissections, 479 retroperitoneal lymph node dissections, 336 adrenalectomies and 108 procedures listed as other. Tumor seeding was reported in 13 cases (0.1%), including 3 nephroureterectomies for transitional cell carcinoma, 4 nephrectomies (incidental transitional cell carcinoma), 4 adrenalectomies for metastases, 1 retroperitoneal lymph node dissection for testicular cancer and 1 pelvic lymph node dissection for cancer of the penis. Port seeding occurred in 10 cases (0.09%) and peritoneal spread in 3 cases (0.03%). CONCLUSIONS: The incidence of tumor seeding after laparoscopic oncological surgery is rare and does not appear greater than what has been historically reported for open surgery. Tumor seeding seems to be most commonly related to the removal of high grade tumors and deviation from oncological surgical principles. PMID- 15126776 TI - Adrenal metastases in 1635 patients with renal cell carcinoma: outcome and indication for adrenalectomy. AB - PURPOSE: Routine removal of the ipsilateral adrenal gland in patients with renal cell carcinoma who undergo nephrectomy has been a matter of dispute. In a retrospective study we screened for subgroups of patients with renal cell carcinoma from a large single center patient population who may have benefited from ipsilateral adrenalectomy. MATERIALS AND METHODS: Radical nephrectomy was performed in 1635 patients at a single institution between 1980 and 2000. A total of 1010 patients underwent radical nephrectomy plus ipsilateral adrenalectomy, whereas in 625 no simultaneous adrenalectomy was performed. Numerous clinical and histopathological parameters were investigated by univariate and multivariate statistical methods for their predictive value in regard to cancer specific survival. RESULTS: Metastases in the adrenal gland were found in 5.5% of patients (56 of 1010) undergoing nephrectomy with adrenalectomy. Of 30 patients with adrenal metastasis and preoperative computerized tomography/magnetic resonance imaging 23 were found to have histological evidence of cancer, approaching a false-negative rate of 23.3%. All patients with false-negative computerized tomography/magnetic resonance imaging had a primary tumor of greater than 4 cm. Patients with adrenal metastases predominately had pT3 or greater tumor stage (82%). Cancer specific survival rates (75% vs 73% for adrenalectomy vs no adrenalectomy) and postoperative complications rates (7% vs 8%) did not differ significantly between the 2 groups. The prognosis in patients with a solitary adrenal metastasis (18 of 56) was more favorable than in patients with additional metastatic sites (38 of 56). CONCLUSIONS: Adrenal metastases from primary renal cell carcinoma were found significantly more often in patients with advanced tumor stages. Ipsilateral adrenalectomy should be recommended for all resectable renal cell carcinoma with a primary tumor of greater than 4 cm or with nonorgan confined tumor stages (T3 or greater) since a false-negative rate of about 20% can be expected with current imaging techniques. PMID- 15126777 TI - Young age is an independent prognostic factor for survival of sporadic renal cell carcinoma. AB - PURPOSE: Compared to older adults with renal cell carcinoma (RCC) our subjective impression has been that younger adults present with more unfavorable histological features and yet respond more favorably to aggressive therapies. We reviewed our experience to validate these observations. MATERIALS AND METHODS: We reviewed the medical records of 106 consecutive patients 40 years or younger and 145 consecutive 58 to 61-year-old patients referred for the surgical management of sporadic RCC between 1992 and 2002. Using univariate and multivariate analyses the pathological characteristics and outcome of the 2 groups were compared. RESULTS: Mean age of the young adults was 34.7 years (range 14 to 40). Of younger adults 24% had tumors with unfavorable features, such as sarcomatoid differentiation, unclassified histology, medullary carcinoma and collecting duct carcinoma, compared with 12% of older adults (p <0.02). However, older adults were more likely to have tumors of advanced local pathological stage (pT3a or greater) (46% vs 31%, p <0.04). Whereas young adults had a higher incidence rate of lymph node metastases at presentation (25% vs 15%, p <0.02), the rate of distant metastatic disease at presentation in young (34%) and older (28%) patients did not differ significantly (p = 0.33). Young age was independently associated with a higher 5-year actuarial disease specific survival rate on multivariate analysis at a median followup of 37 months (66% vs 52%, adjusted HR 2.64, 95% CI 1.45 to 4.79, p <0.002). On multivariate analysis of patients without distant metastases at presentation young adults also had improved recurrence-free survival (median time to recurrence 32.4 vs 23.5 months, HR 2.23, 95% CI 1.04 to 4.78, p <0.04). CONCLUSIONS: Young adults with RCC were more likely to have unfavorable histological features and a higher incidence of lymph node metastases than an older cohort of adults. Despite these differences on multivariate analysis young patients had improved disease-specific and recurrence free survival following treatment. Whether age specific differences in host-tumor interaction exist in patients with RCC deserves further study. PMID- 15126778 TI - Expression of hepatocyte growth factor and its receptor C-met in acquired renal cystic disease associated with renal cell carcinoma. AB - PURPOSE: The development of renal cell carcinoma (RCC) is a critical problem in acquired renal cystic disease (ARCD) in patients with end stage renal disease, particularly males on chronic hemodialysis. We examined the expression of hepatcyte growth factor (HGF) and its receptor c-met in ARCD associated with RCC to elucidate the role of the HGF/c-met pathway in renal cyst development and subsequent tumor transformation. MATERIALS AND METHODS: Immunohistochemical expression of HGF, c-met and Bcl-2 was examined in 15 normal tissue samples of kidneys obtained from nephrectomy for localized RCC and 19 with ARCD associated with RCC from 16 patients on dialysis. The expression of HGF mRNA was examined by reverse transcriptase-polymerase chain reaction. RESULTS: In normal kidneys immunostaining for HGF was scarcely detected in renal tubular cells. Bcl-2 and c met immunostaining was observed mainly in distal tubular cells. In nontumor areas of kidneys with ARCD associated RCC the expression of HGF and c-met was up regulated in tubular and the cyst epithelial cells. Hyperplastic cysts with multilayer epithelium and micropapillary projection were the predominant cysts stained with HGF and c-met. Immunostaining for HGF and c-met was also detected in RCC regions. The Bcl-2 immunostaining pattern was similar to that of HGF and c met in nontumor and RCC regions. On reverse transcriptase-polymerase chain reaction HGF mRNA expression was up-regulated in nontumor areas and RCC regions in ARCD. CONCLUSIONS: These results suggest that up-regulation of HGF and its receptor c-met may be involved in renal cyst formation and subsequent tumor transformation in patients with end stage renal disease. Increased Bcl-2 expression may promote this process through the inhibition of apoptosis. PMID- 15126779 TI - High expression of chemokine gene as a favorable prognostic factor in renal cell carcinoma. AB - PURPOSE: The presence and potential role of chemokines in clinical tumors remain poorly understood. Chemokines are a large family of chemoattractant cytokines with several members that are also able to regulate angiogenesis. We hypothesized that chemokines may have an important role in regulating tumor growth in renal cell carcinoma (RCC). To begin to test this hypothesis chemokine gene expression and its influence on prognosis, cellular infiltration and angiogenesis in RCC were examined. MATERIALS AND METHODS: A total of 51 patients with sporadic RCC were analyzed for the expression of the 7 chemokine genes interleukin-8, gamma interferon inducible protein-10 (IP-10), monokine induced by gamma-interferon (MIG), macrophage chemoattractant protein-1, macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and regulated and activated normal T cells excreted and secreted (RANTES) using semiquantitative reverse transcriptasepolymerase chain reaction. Lymphocyte infiltration and microvessel density were determined immunohistochemically by staining CD8 and CD34 cells, respectively. RESULTS: : The expression of IP-10, MIG, MIP-1beta and RANTES was significantly increased in tumor compared to normal kidney tissues. The expression of IP-10, MIG and MIP 1beta showed an inverse correlation with tumor size. Stages 1 to 3 tumors expressing high levels of IP-10, MIG, MIP-1beta and RANTES did not recur after curative surgery. Intratumor expression of IP-10, MIG and MIP-1beta showed a positive correlation with the degree of CD8 cell infiltrating in the tumor and an inverse correlation with microvessel density. CONCLUSIONS: These results suggest that IP-10, MIG and MIP-1beta are expressed at high levels in tumors that rarely recur after surgery. The antitumorigenic effect of IP-10, MIG and MIP-1beta may result from the recruitment of lymphocyte infiltration and/or inhibition of angiogenesis in RCC. The amplification of chemokine expression by immunotherapy or gene therapy may be a practical and effective strategy to promote tumor regression. PMID- 15126780 TI - C-kit expression in sarcomatoid renal cell carcinoma: potential therapy with imatinib. AB - PURPOSE: Sarcomatoid (S) renal cell carcinoma (RCC) is an uncommon subtype of RCC with a poor prognosis because of its local aggressiveness and high metastatic rate. Currently, there is no specific, effective treatment for it. A relatively nontoxic tyrosine kinase inhibitor, imatinib (STI-571) has been approved as a target therapy in neoplasms that express c-Kit. We investigated c-Kit expression in this type of tumor, which to our knowledge has not been previously described. MATERIALS AND METHODS: We reviewed 215 cases of RCC diagnosed at our department from 1995 to 2002. Of the cases 20 (9.3%) were SRCC. Formalin fixed, paraffin embedded material was available in 19 cases. We performed immunohistochemical staining against c-Kit using rabbit polyclonal antihuman antibody (CD117, Dako Corp., Carpinteria, California), diluted 1:100. Its expression was evaluated in the epithelial and the spindle components. RESULTS: Two of the 20 SRCC cases (10%) showed no epithelial differentiation. The epithelial component was conventional RCC in 10 cases (50%), papillary RCC in 5 (25%) and chromophobe RCC in 3 (15%). A total of 16 cases (80%) presented at an advanced stage at diagnosis, namely T3 or T4 and/or metastatic disease. Immunohistochemical study showed positivity in the epithelial component only in the 3 chromophobe SRCCs. The sarcomatoid component was positive for c-Kit in 18 cases (94.7%). CONCLUSIONS: High c-Kit expression in SRCC in our series and the existence of a target therapy, imatinib (STI-571), against cells that express this receptor open the possibility of using this treatment for these tumors, especially in cases of advanced disease. PMID- 15126781 TI - Safety and efficacy of partial nephrectomy for all T1 tumors based on an international multicenter experience. AB - PURPOSE: We compared cancer specific survival of patients undergoing partial and radical nephrectomies for T1N0M0 renal tumors according to tumor size in a large multicenter series. MATERIALS AND METHODS: A retrospective analysis of 1454 patients undergoing partial or radical nephrectomy for T1N0M0 renal tumors from 7 international academic centers was performed. Data were obtained for each patient including TNM stage (determined according to the 2002 TNM criteria), tumor size, type of surgery (partial versus radical nephrectomy) and cancer specific survival. Recurrence events were recorded when available. RESULTS: Partial and radical nephrectomies were performed in 379 (26.1%) and 1075 (73.9%) cases, respectively. Mean followup +/- SD was 62.5 +/- 51.8 months. Recurrence data were available on 544 patients. There were no significant differences in local or distant recurrence rates between patients undergoing partial or radical nephrectomy for either T1a (p = 0.6) or T1b tumors (p = 0.5). For patients with T1a tumors, there was no significant difference in the rate of cancer specific deaths between the partial (314) and radical (499) nephrectomy groups (2.2% versus 2.6%, respectively, p = 0.8). For patients with T1b tumors there was also no significant difference in the rate of cancer specific deaths between patients undergoing partial (65) and patients undergoing radical (576) nephrectomy (6.2% versus 9%, respectively, p = 0.6). CONCLUSIONS: Partial nephrectomy is becoming the gold standard for renal tumors less than 4 cm but this treatment is much more controversial for larger T1 tumors. This large multicenter study suggests that it is safe to expand the indications of partial nephrectomy to include patients with T1N0M0 tumors up to 7 cm. However, careful patient selection remains necessary. PMID- 15126782 TI - 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. AB - PURPOSE: We determined if 1 immediate instillation of chemotherapy after transurethral resection (TUR) decreases the risk of recurrence in patients with stage Ta T1 single and multiple bladder cancer overall and separately. MATERIALS AND METHODS: A meta-analysis was performed of the published results of randomized clinical trials comparing TUR alone to TUR plus 1 immediate instillation of chemotherapy. RESULTS: Our study included 7 randomized trials with recurrence information on 1476 patients. Based on a median followup of 3.4 years and a maximum of 14.5 years, 267 of 728 patients (36.7%) receiving 1 postoperative instillation of epirubicin, mitomycin C, thiotepa or (2'R)-4'-O-tetrahydropyranyl doxorubicin (pirarubicin) had recurrence compared to 362 of 748 patients (48.4%) with TUR alone, a decrease of 39% in the odds of recurrence with chemotherapy (OR 0.61, p <0.0001). Patients with a single tumor (OR 0.61) and those with multiple tumors (OR 0.44) benefited. However, after 1 instillation 65.2% of patients with multiple tumors had recurrence compared to 35.8% of patients with single tumors, showing that 1 instillation alone is insufficient treatment for patients with multiple tumors. CONCLUSIONS: One immediate intravesical instillation of chemotherapy significantly decreases the risk of recurrence after TUR in patients with stage Ta T1 single and multiple bladder cancer. It is the treatment of choice in patients with a single, low risk papillary tumor and is recommended as the initial treatment after TUR in patients with higher risk tumors. PMID- 15126783 TI - How to avoid false-negative dynamic sentinel node procedures in penile carcinoma. AB - PURPOSE: Evaluation of the false-negative dynamic sentinel node procedures in penile carcinoma at our institute. MATERIALS AND METHODS: Between January 1994 and February 2003, 123 patients with penile squamous cell carcinoma underwent dynamic sentinel node biopsy. RESULTS: The sentinel node revealed metastasis in 28 (23%) of 123 patients. Regional recurrence after excision of a tumor-negative sentinel node or after nonvisualization was seen in 6 patients resulting in a false-negative rate of 18% (6 of 34). We assume that 1 false-negative case was due to tumor blockage, 3 to tumor blockage and rerouting, 1 to a pathological sampling error and 1 to a low radioactivity level in the sentinel node during surgery. CONCLUSIONS: Based on the false-negative results, important adaptations have been made in the dynamic sentinel node biopsy procedure for penile carcinoma at our institute. Pathological analysis was extended by serial sectioning and immunohistochemical staining, and preoperative ultrasonography with fine needle aspiration cytology has been added. Furthermore, exploration of groin without visualized sentinel nodes and intraoperative palpation of the wound have been introduced. PMID- 15126784 TI - Quantitative GSTP1 methylation levels correlate with Gleason grade and tumor volume in prostate needle biopsies. AB - PURPOSE: Methylation of the GSTP1 gene promoter region is the most common epigenetic change in prostate cancer. The quantitative GSTP1 methylation assay, a recently developed methylation specific and polymerase chain reaction based technique, allows the accurate discrimination of benign prostate tissue and prostate cancer even in small, formalin fixed prostate needle biopsies. In this study we investigated whether GSTP1 methylation correlated with Gleason grade and tumor volume in prostate biopsies. We also compared quantitative GSTP1 methylation with the histological diagnosis in each biopsy. MATERIALS AND METHODS: Formalin fixed, paraffin embedded prostate needle cores (25 mu) were used for the GSTP1 methylation study. After excluding 98 prostate needle cores (47%) with insufficiently extracted DNA 109 biopsies from 40 patients with prostate cancer of various Gleason grades and tumor volume, and benign prostate tissue were tested for GSTP1 methylation using quantitative fluorogenic real-time methylation specific polymerase chain reaction. RESULTS: A total of 13 benign and 96 cancer biopsy parts were included in this study. GSTP1 methylation was positive (GSTP1-to-ACTB ratio 5 or greater) in 83 of 96 cancer parts (86.5%). All 13 benign parts (100%) were negative for GSTP1 methylation with a methylation level of 0 in 12 and 1 in 1. The sensitivity and specificity of quantitative GSTP1 methylation assay were 86.5% and 100%, respectively. Quantitative GSTP1 methylation correlated with Gleason grade, demonstrating higher GSTP1 methylation in higher Gleason grade tumors (Spearman correlation coefficient 0.377, p < 0.0001). Likewise, GSTP1 methylation also correlated with the cancer percent per biopsy (Spearman correlation coefficient 0.567, p < 0.0001). A multiple linear regression model predicted the greater contribution of cancer percent than Gleason grade to quantitative GSTP1 methylation levels. CONCLUSIONS: We confirmed that the quantitative GSTP1 methylation assay reliably discriminates benign and malignant prostate tissues, thus, augmenting the routine histological evaluation of prostate needle biopsies. In addition, quantitative GSTP1 methylation also correlated with prostate cancer Gleason grade and cancer volume, suggesting that quantitative GSTP1 methylation may be of prognostic significance. PMID- 15126785 TI - Lower body mass index is associated with a higher prostate cancer detection rate and less favorable pathological features in a biopsy population. AB - PURPOSE: Body mass index (BMI), calculated as weight in kg divided by the square of height in m, is used as an indicator of obesity. We assessed the relationship between BMI, and prostate cancer detection rates and biopsy features in a referral based biopsy population. MATERIALS AND METHODS: A total of 787 consecutive patients referred for abnormal digital rectal examination and/or prostate specific antigen (PSA) greater than 4 ng/ml underwent systematic prostate biopsy. Three standard categories of BMI were considered, namely normal less than 25, overweight-25 to 29.9 and obese-30 or greater kg/m. The presence or absence of cancer, percent of core involvement and tumor grade were correlated with BMI. Additional analyses controlled for patient age, PSA and prostate volume. RESULTS: For the entire population detection rates were highest in the normal BMI group compared to the overweight or obese group (52% vs 37% vs 42%, p = 0.0026). When stratified by age, this observation was true for men younger than 70 years (49% vs 32% vs 37%, p = 0.0042) but not for men 70 years or older. When only patients with PSA 10 ng/ml or less were considered, detection rates were highest in the normal BMI group (44% vs 28% vs 36%, p = 0.0061). This observation also persisted in patients younger than 70 years with PSA 10 ng/ml or less, or when only patients younger than 70 years with a total prostate volume of less than 50 cc were included. Of patients with cancer those with a normal BMI had a greater length of needle core involvement on biopsy. CONCLUSIONS: Normal BMI correlates with a higher cancer detection rate and larger cancers in men undergoing prostate biopsy. PMID- 15126786 TI - A comparison of extended biopsy and sextant biopsy schemes for predicting the pathological stage of prostate cancer. AB - PURPOSE: We compared the performance of the extended multisite directed biopsy strategy to the sextant component of this strategy for predicting the pathological stage and Gleason score of the radical prostatectomy specimen. MATERIALS AND METHODS: We studied 157 men in whom prostate cancer was diagnosed by extended multisite directed biopsy and who underwent radical retropubic prostatectomy. The pretreatment variables of serum prostate specific antigen, prostate specific antigen density, biopsy specimen Gleason score, the location, number and percent of cancer containing cores, greatest tumor length in a single core and greatest percent of tumor in a single core were determined and compared with the pathological features of prostate cancer in the radical prostatectomy specimens. A comparison of the information obtained from sextant component cores of the extended biopsy strategy with that from all cores of the extended biopsy strategy was performed using chi-square statistics and ROC curve analysis. RESULTS: When comparing the areas under the ROC curves, the extended multisite directed biopsy strategy was found to have greater predictive power for extraprostatic extension than the sextant core component of this biopsy scheme, although the difference was not significantly different. The sextant component was equivalent to the extended biopsy strategy for predicting the prostatectomy specimen Gleason score. CONCLUSIONS: The extended biopsy strategy has better performance in the upper sensitivity ranges compared to the sextant technique for predicting extraprostatic extension. PMID- 15126787 TI - The percent of biopsy cores positive for cancer is a predictor of advanced pathological stage and poor clinical outcomes in patients treated with radical prostatectomy. AB - PURPOSE: We examined if the percent of positive biopsies is associated with features of biologically aggressive prostate cancer, biochemical progression and development of distant metastases in patients undergoing radical prostatectomy (RP). MATERIALS AND METHODS: Multivariate analyses of preoperative features in 605 consecutive patients who underwent RP for clinically localized disease were evaluated to determine the association between the percent positive biopsy cores (PosBx), pathological stage and grade, and biochemical progression following RP. The percent of PosBx cores was defined using the formula, (number of positive biopsy cores/total number of biopsy cores) x 100. RESULTS: The mean number of biopsy cores and percent PosBx cores +/- SE was 8.8 +/- 6.0 and 31.4 +/-21.1, respectively. Higher percent PosBx was significantly associated with higher preoperative prostate specific antigen (PSA), extracapsular extension, seminal vesicle invasion, positive surgical margins, higher final Gleason sum, lymphovascular invasion, perineural invasion and metastases to regional lymph nodes. On multivariate analyses adjusted for the effects of standard preoperative features percent PosBx was associated with nonorgan confined disease, seminal vesicle invasion and biochemical progression after surgery (p = 0.049, 0.050 and 0.006, respectively). Percent PosBx retained its independent association with PSA progression after adjustment for the effects of postoperative pathological features (p = 0.015). Higher percent PosBx was associated with shorter PSA doubling time after PSA progression, and an increased risk of distant metastases and overall mortality (p = 0.039, 0.001 and 0.018, respectively). CONCLUSIONS: Percent PosBx is associated with established pathological features, biochemical progression, distant metastases and overall death in patients who undergo RP for clinically localized disease. Percent PosBx should be included in preoperative predictive models for prognosticating outcomes after primary treatment and it may assist in selecting patients for inclusion in neoadjuvant and/or adjuvant therapy trials. PMID- 15126788 TI - Preoperative model for predicting prostate specific antigen recurrence after radical prostatectomy using percent of biopsy tissue with cancer, biopsy Gleason grade and serum prostate specific antigen. AB - PURPOSE: We developed a preoperative model to risk stratify patients for prostate specific antigen (PSA) failure following radical prostatectomy (RP) and identify those at high risk who would be potential candidates for neoadjuvant clinical trials. MATERIALS AND METHODS: A retrospective survey of 459 patients from the SEARCH Database treated with RP between 1990 and 2002 was done. Multivariate analysis was used to compare the preoperative variables of patient age, race, PSA, biopsy Gleason score, clinical stage and percent of prostate needle biopsy tissue with cancer for the ability to predict time to PSA recurrence following RP. Significant independent predictors were combined to create a novel risk grouping model. RESULTS: On multivariate analysis biopsy Gleason score (p < 0.001), percent of biopsy tissue with cancer (p < 0.001) and serum PSA (p = 0.001) were the only significant independent predictors of PSA failure. Combining these 3 significant predictors of PSA failure using previously published cutoff points for each variable generated a 4 tier preoperative model for predicting biochemical failure following RP (HR 1.91 for each 1 risk category increase, CI 1.62 to 2.26, p < 0.001). The model further stratified patients who were already stratified into low, intermediate and high risk groups based on a previously described model using PSA, biopsy Gleason score and clinical stage. A simplified table was developed to predict the risk of biochemical recurrence within 2 years following surgery, as stratified by percent of tissue with cancer, PSA and biopsy Gleason score. CONCLUSIONS: A combination of serum PSA, biopsy Gleason score and percent of prostate biopsy tissue with cancer define a new preoperative model for predicting PSA failure following RP. This model further stratified patients who were already stratified based on PSA, biopsy Gleason score and clinical stage, and it can be used preoperatively to identify patients at high risk who would be candidates for neoadjuvant clinical trials. Using this model an easy to use table was developed to predict preoperatively the 2-year risk of PSA recurrence following RP. PMID- 15126789 TI - Validation of increasing prostate specific antigen as a predictor of prostate cancer death after treatment of localized prostate cancer with surgery or radiation. AB - PURPOSE: Many patients who undergo surgery or radiation therapy to treat localized prostate cancer experience an increase in serum prostate specific antigen (PSA) after treatment. This study documents patterns of PSA recurrence after surgery or radiation to treat localized prostate cancer and quantifies the extent to which an increasing PSA predicts death from prostate cancer. MATERIALS AND METHODS: Posttreatment PSA levels were measured on a population based cohort of 1136 men diagnosed with localized prostate cancer in community practice in Connecticut between 1990 and 1992, and treated within 6 months of diagnosis with surgery or radiation with or without androgen withdrawal therapy. The major outcome measure was death from prostate cancer. RESULTS: PSA recurrence followed a log-linear pattern over time. Patients who died of prostate cancer had a median PSA doubling time of 0.8 years (25th and 75th percentiles 0.5 to 1.4 years). Patients who did not die of prostate cancer within 10 years of diagnosis had either no posttreatment increase in serum PSA (40%) or had a PSA doubling time longer than 1 year (44%). CONCLUSIONS: Patients whose posttreatment PSA doubling times before the initiation of androgen withdrawal therapy are less than 1 year are at high risk of dying of prostate cancer within 10 years of diagnosis. Men with PSA recurrences that are doubling at rates greater than 1 year are at low risk of death from prostate cancer within 10 years of diagnosis. PMID- 15126790 TI - Differentiation of benign prostatic hyperplasia from prostate cancer using prostate specific antigen dynamic profile after transrectal prostate biopsy. AB - PURPOSE: Serum prostate specific antigen (PSA) level is increased after needle biopsy (Bx) of the prostate. This study tested the hypothesis that the prostate harboring malignant lesions demonstrates less leakage of PSA after prostate biopsy and this phenomenon can be helpful in discriminating benign from cancer diagnosis. MATERIALS AND METHODS: This prospective study was divided into 3 separate phases. Sextant prostate biopsy was done with transrectal ultrasound guidance, and the change and PSA values after biopsy were evaluated. Phases 1 and 2 had 20 and 41 patients, respectively. PSA dynamic profiles were plotted. We defined the most appropriate timing for blood sampling and the cutoff value of the PSA ratio (post-Bx total PSA-to-pre-Bx total PSA) to be applied for further assessment. Phase 3 recruited 97 cases, of which 66 satisfied the end point criteria in which a diagnosis such as malignancy, or 3 successive benign biopsies or benign transurethral resection of prostate was obtained. RESULTS: From phases 1 and 2 the cutoff value for the PSA ratio was 2.0, and the timing of blood sampling was 60 minutes after the biopsy. In phase 3 of those whose PSA ratio was less than 2.0, 92.6% (25) had cancer. For those whose PSA ratio was greater than 2.0, 82.1% (32) were benign. Of the 4 cases with a PSA ratio less than 2.0 and an initial benign biopsy, 3 (75%) were proven to have cancer later. CONCLUSIONS: Evaluating the 1-hour PSA ratio might be helpful for clinicians to select the high risk patients who might have cancer in the prostate. Repeat biopsy should be suggested for cases with a smaller PSA ratio in spite of initial benign results. PMID- 15126791 TI - Evaluation of serum and seminal levels of prostate specific antigen in men with spinal cord injury. AB - PURPOSE: Recent studies have used prostate specific antigen (PSA) as an indicator of prostate gland activity in patients with spinal cord injury (SCI). Thus, the present study was performed to determine whether SCI can induce alterations in total serum and seminal PSA, and to compare the findings obtained to those of normal men (controls). MATERIALS AND METHODS: A total of 44 men with SCI (cases, mean age +/- SD 33.98 +/- 9.12 years) and 44 controls (mean age +/- SD 34.09 +/- 9.16 years) were studied. Blood and semen samples were collected after 3 days of abstinence from ejaculation and stored at a controlled temperature between -70 and -79C. Seminal fluid was kept at room temperature for 15 minutes before storage. The tests for determination of total serum and seminal PSA were performed using AxSYM equipment and reagents. RESULTS: The mean total seminal PSA obtained from patients (0.609 mg/ml) was lower than the 0.773 mg/ml value obtained from controls (p = 0.0012), but the mean total serum PSA of patients (0.918 ng/ml) did not differ significantly from that obtained from controls (0.976 ng/ml, p = 0.9967). CONCLUSIONS: SCI patients have a significant decrease in total seminal PSA but total serum PSA is not affected by this lesion. PMID- 15126792 TI - Prostate specific antigen testing. PMID- 15126793 TI - Variation in prostate specific antigen results from 2 different assay platforms: clinical impact on 2304 patients undergoing prostate cancer screening. AB - PURPOSE: Urologists must contend with fluctuating prostate specific antigen (PSA) results from different assay platforms when deciding whether prostate biopsy is indicated. The contribution of cross-platform variation to these fluctuations is not well understood. To address this question we performed a cross-sectional study comparing 2 popular PSA assays during prostate cancer screening. MATERIALS AND METHODS: The Baylor College of Medicine Prostate Cancer Screening Program is a community outreach program providing free screening to Houston residents. From September 18th to 23, 2000, 2,304 patients underwent digital rectal examination and parallel serum PSA assay by the Hybritech Access (Beckman Coulter, Inc., Chaska, Minnesota) and Centaur (Bayer Diagnostics, Tarrytown, New York) systems. The Wilcoxon signed ranks test was used to compare results. RESULTS: Median PSA was low for the 2 assays (Centaur 0.99 ng/ml and Access 1.09 ng/ml) and Access results were significantly higher (1.23 times) than those obtained with Centaur (p <0.001). Frozen serum from 50 patients with PSA greater than 2.5 were then re assayed using a third methodology (third Generation Immulite, Diagnostic Products Corp., Los Angeles, California). These data were consistent with Centaur results and significantly lower than Access results (p <0.001). Using a cutoff of PSA greater than 4.0 ng/ml 55 of the 288 patients (19%) with PSA greater than 2.5 ng/ml on the 2 platforms would have been candidates for prostate biopsy based on Access but not on Centaur data. CONCLUSIONS: In a large screening population the Access system measured consistently higher PSA than the Centaur system. These findings provide a basis for interpreting PSA results obtained from 2 commonly used clinical assays. PMID- 15126794 TI - Serum pro-prostate specific antigen preferentially detects aggressive prostate cancers in men with 2 to 4 ng/ml prostate specific antigen. AB - PURPOSE: Pro forms of prostate specific antigen (PSA) have been reported to be more cancer specific markers of prostate cancer than total PSA and they also may preferentially detect the more aggressive forms of the disease. MATERIALS AND METHODS: Research immunoassays with high specificity for pro-PSA forms were used to study 1091 retrospective serum specimens, including 555 with 2 to 4 and 536 with 4 to 10 ng/ml PSA, from men enrolled in prostate cancer screening studies who underwent prostate biopsy. RESULTS: In the 2 to 4 ng/ml PSA range the ratio of pro- to free-PSA (percent pro-PSA) using a cutoff of 1.8% for recommending prostate biopsy detected 90% of cancers, including 16 of 16 extracapsular tumors and 28 of 29 tumors with a pathology Gleason score of 7 or greater, while avoiding 19% of unnecessary biopsies. Serum percent pro-PSA was significantly increased for Gleason score 7 or greater vs less than 7 (p = 0.0018). In the PSA range of 4 to 10 ng/ml percent pro-PSA had the highest cancer specificity, avoiding 31% of unnecessary biopsies, while detecting 34 of 35 cancers with a pathology Gleason score of 7 or greater and 29 of 31 extracapsular tumors. Neither percent free PSA nor complexed PSA enhanced the detection of aggressive cancers in the 4 to 10 ng/ml PSA range. CONCLUSIONS: Percent pro-PSA was superior to percent free and calculated complexed PSA for the detection of prostate cancer in the PSA range of 2 to 10 ng/ml and it had selectivity for detecting more aggressive cancers, as indicated by Gleason score 7 or greater and/or extracapsular tumor extension. PMID- 15126795 TI - Prostate cancer detection in the prostate specific antigen range of 2.0 to 3.9 ng/ml: value of percent free prostate specific antigen on tumor detection and tumor aggressiveness. AB - PURPOSE: We evaluated the positive predictive value and cancer detection rate in the prostate specific antigen (PSA) range of 2.0 to 3.9 ng/ml and assessed the value of percent free (F) PSA (FPSA) on tumor detection and tumor aggressiveness in this low PSA range. MATERIALS AND METHODS: Of 3623 men who were attending the second round of screening within the European Randomized Study of Screening for Prostate Cancer, section Rotterdam 883 had PSA values of 2.0 to 3.9 ng/ml. These men were offered laterally directed sextant biopsy. FPSA was prospectively determined from pretreatment serum. Cancers were classified as prognostically favorable and unfavorable using biopsy results and other pretreatment diagnostic features. RESULTS: Using the PSA range of 2.0 to 3.9 ng/ml as a biopsy indication 126 cancers were detected, resulting in a positive predictive value of 17.1% and a cancer detection rate of 14.3%. By using percent FPSA and setting relative sensitivity at 95% 9% of biopsies could have been avoided. Unfavorable tumor characteristics were found in 46.9% of the men with T1C tumors. Mean percent FPSA was significantly lower in such men compared to men with favorable tumor characteristics. Of the men with percent FPSA lower than 10% 90% had unfavorable tumor characteristics. CONCLUSIONS: The PSA range 2.0 to 3.9 ng/ml is accessible for prostate cancer screening. Percent FPSA is of moderate value in avoiding unnecessary biopsies in the PSA range of 2.0 to 3.9 ng/ml. However, when assessing tumor aggressiveness in biopsy results, percent FPSA is predictive and can be used to select treatment options, such as watchful waiting. PMID- 15126796 TI - The economic burden of metastatic and prostate specific antigen progression in patients with prostate cancer: findings from a retrospective analysis of health plan data. AB - PURPOSE: We evaluated the economic burden of metastatic and prostate specific antigen (PSA) progression in patients with prostate cancer (CaP) using a cancer registry linked administrative database. MATERIALS AND METHODS: A retrospective cohort evaluation of 2056 patients with CaP was done at Henry Ford Health System from 1995 to 2000. Records were examined for metastatic progression via International Classification of Disease-9-CM codes for metastasis and for PSA progression using accepted definitions based on initial therapy type. Health care resource charges 6 months and 1 year before and after progression were compared using pairwise t tests. A generalized linear model determined the effect of progression on charges and compared initial care, continuing care and terminal care charges in the progressed and nonprogressed groups, while controlling for baseline covariates (stage and age). RESULTS: Patients with CaP had a mean age of 68 years, were mostly white (52%), had localized (88%) and moderately differentiated (66%) tumors, and a median baseline PSA of 7.0 ng/ml. Of patients 8.9% had metastatic progression at a mean followup of 3.6 years, while 16.1% had PSA progression at 4.5 years. After controlling for baseline covariates metastatic progression resulted in significant increases in charges (US dollars 92523 vs US dollars 58036, p < 0.0001). PSA progressed patients incurred significantly higher charges than nonprogressed patients (US dollars 69321 vs US dollars 58351, p = 0.0039), controlling for followup time, baseline stage, grade and treatment. CONCLUSIONS: In CaP cases metastatic and PSA progression pose a significant economic burden irrespective of baseline stage, grade and treatment. Treatments that slows or prevents meta-static and PSA progression could offset this cost. PMID- 15126797 TI - Validation of the Kattan preoperative nomogram for prostate cancer recurrence using a community based cohort: results from cancer of the prostate strategic urological research endeavor (capsure). AB - PURPOSE: The Kattan preoperative nomogram combines preoperative prostate specific antigen (PSA), biopsy Gleason grade and clinical stage to estimate disease recurrence after radical prostatectomy. Several studies using patient data from academic centers have validated the nomogram. We assessed the performance of the Kattan nomogram using the Cancer of the Prostate Strategic Urological Research Endeavor database, a national, largely community based observational disease registry. MATERIALS AND METHODS: From the Cancer of the Prostate Strategic Urological Research Endeavor database we identified 1701 men with clinically localized prostate cancer undergoing radical prostatectomy with sufficient pretreatment information and PSA followup after surgery. Disease recurrence was defined as 2 consecutive PSA values 0.2 ng/ml or greater, or a second cancer treatment more than 6 months after prostatectomy. A concordance index was used to evaluate the performance of the nomogram compared to observed 5-year recurrence free survival (Kaplan-Meier). Kattan nomogram scores were calculated for each patient and stratified into 6 groups for analysis. RESULTS: In our cohort of 1701 men 413 (24%) had evidence of disease recurrence. Median followup in these patients was 2.3 years. Kattan nomogram scores were 17% to 99% (mean 79%). The overall concordance index was 0.68. Varying the definition of recurrent disease and excluding patients with imputed data did not substantially alter nomogram performance (concordance index 0.65 to 0.70). The Kattan nomogram tended to overestimate 5-year freedom from recurrence in patients with scores of 65% and higher. CONCLUSIONS: We noted the reasonable performance of the Kattan nomogram for predicting cancer outcomes after radical prostatectomy using a community based population. Although concordance is lower than in previous validation studies and the nomogram overestimates recurrence-free survival in patients at lower risk, the model is fairly robust and it provides important information when counseling patients regarding treatment options in the community setting. Further refinements in pretreatment estimation of disease-free survival and ultimately overall survival are needed. PMID- 15126798 TI - Predictors of metastatic disease in men with biochemical failure following radical prostatectomy. AB - PURPOSE: We determined clinical and pathological predictors of positive bone scans and computerized tomography (CT) in patients with biochemical recurrence after radical prostatectomy (RP). MATERIALS AND METHODS: A retrospective analysis of patients treated with RP at West Los Angeles Veterans Affairs Medical Center and University of California-Los Angeles Medical Center was performed to identify men with biochemical recurrence. All postoperative bone scans and pelvic CT following recurrence and prior to the initiation of hormone ablation therapy were reviewed. Preoperative clinical variables, pathological findings, serum prostate specific antigen (PSA) at postoperative imaging and postoperative PSA doubling time were compared between patients with positive and negative imaging study results. RESULTS: A total of 128 patients with biochemical recurrence after RP who had postoperative pelvic CT or bone scans available were identified. A total of 97 bone scans were obtained, of which 11 (11%) were positive, and 71 CT scans were obtained, of which 5 (7%) were positive. Men with PSA doubling time less than 6 months were at increased risk of a positive bone scan (26% vs 3%) or positive CT (24% vs 0%) relative to men with longer PSA doubling time. In men with PSA doubling time less than 6 months the risk of a positive study highly depended on PSA at the time of imaging. In men with PSA less than 10 ng/ml the incidence of a positive study was 0% for pelvic CT and 11% for bone scan. In men with PSA greater than 10 ng/ml the risk of a positive study was 57% for pelvic CT and 46% for bone scan. In men with PSA doubling time greater than 6 months no clear relationship to PSA was seen, although the number of patients with a positive study was extremely low (positive bone scans 3% and positive CT 0%). However, none of the 6 imaging studies performed in men with PSA doubling times greater than 6 months and a markedly elevated PSA of 20 to 90 ng/ml was positive. CONCLUSIONS: The risk of detecting metastatic disease by bone scan or pelvic CT in men with biochemical recurrence following RP with PSA doubling time greater than 6 months is low despite marked PSA increases up to 90 ng/ml. In men with PSA doubling time less than 6 months the risk of detecting metastatic disease markedly increases when PSA is greater than 10 ng/ml. These results have important implications for the timing of imaging in patients with biochemical recurrence following RP. PMID- 15126799 TI - Transrectal focused ultrasound combined with transurethral resection of the prostate for the treatment of localized prostate cancer: feasibility study. AB - PURPOSE: We studied the feasibility of combined treatment with high intensity focused ultrasound (HIFU) and transurethral resection or incision of the prostate for localized prostate cancer to decrease the risk of posttreatment prolonged urinary retention. MATERIALS AND METHODS: Included in this feasibility study were 30 patients fulfilling certain criteria, namely localized prostate cancer indicated for HIFU, age 60 years or older, prostate volume 45 cc or less, no more than 4 positive samples at sextant biopsy, baseline prostate specific antigen (PSA) 10 ng/ml or less and no evidence of cancer extension. They received the combined treatment under general anesthesia using an Ablatherm HIFU device (EDAP SA, Lyon, France). RESULTS: The enrolled patients were a mean of 72 years old and presented with a median prostate volume of 30 cc. Median Gleason score was 6, median PSA was 7 ng/ml and pretreatment sextant biopsies provided a median of 2 positive samples. Mean operative time was 2 hours 48 minutes, including resection and HIFU. An average of 616 HIFU shots were delivered. The urinary catheter was removed at day 2 after treatment. Median hospital stay was 3 days. Only a few complications were observed. In regard to the oncological aspects at a mean of 20 months of followup 86% of the patients had negative biopsies after HIFU. Median PSA was 0.9 ng/ml. At 1 year of followup the mean International Prostate Symptom Score was 8. Regarding sexual function, 73% of previously potent patients reported preserved sexual activity. CONCLUSIONS: The combination of endoscopic resection or incision of the prostate with HIFU treatment decreases urinary catheterization time and improves posttreatment urinary status without additional morbidity. PMID- 15126800 TI - Validity of sentinel lymph node concept for patients with prostate cancer. AB - PURPOSE: We investigated the feasibility and validity of the sentinel lymph node concept for patients with prostate cancer. MATERIALS AND METHODS: A total of 24 patients (mean age 68.1 years) with prostate cancer but without detectable distant metastases were enrolled in this study. Radioactive tracer (technetium labeled phytate) was injected into the prostate 5 to 6 hours before surgery. Preoperative lymphoscintigraphy was viewed, and in vivo and ex vivo gamma probing was performed during the surgery, which included backup extended lymph node dissection. Radioactivity of the excised lymph nodes was measured and validated with an autowell scintillation counter. RESULTS: Radioactive positive (hot) nodes were identified in 17 of the 24 (70.8%) patients by lymphoscintigraphy, in 21 of 24 (87.5%) patients by in vivo probing and in 23 of 24 (95.8%) patients by ex vivo probing. On an individual basis the numbers of hot nodes identified by lymphoscintigraphy and in vivo probing were low (40% and 36%, respectively) compared to the high detection rate for ex vivo probing (91%). CONCLUSIONS: Although sentinel lymph nodes can be used for prostate cancer, detailed lymph node mapping and extended dissection backup are currently needed. It is also important to establish novel procedures for accurate and complete removal of all hot nodes. PMID- 15126801 TI - Bicalutamide 150 mg maintains bone mineral density during monotherapy for localized or locally advanced prostate cancer. AB - PURPOSE: We evaluated changes in bone mineral density (BMD), fat-free mass (FFM) and serum lipid levels during bicalutamide 150 mg monotherapy compared with medical castration for 2 years. MATERIALS AND METHODS: A total of 103 men with localized or locally advanced prostate cancer (T1-T4, Nx, M0) for whom immediate androgen deprivation was indicated were enrolled in this prospective, multicenter, open-label, parallel group study. Patients were randomized to bicalutamide 150 mg once daily (51) or medical castration with a luteinizing hormone releasing hormone analogue (52) for 96 weeks. Primary end points were mean percent change from baseline in lumbar spine BMD, hip BMD and FFM at 96 weeks. Mean changes in lipid parameters with time were also evaluated. RESULTS: BMD was maintained during bicalutamide 150 mg monotherapy (+2.42% for lumbar spine BMD and +1.13% for hip BMD at week 96), while castration was associated with a progressive loss in BMD (-5.40% and -4.39% at week 96, respectively, both p <0.0001 at week 96). There was no significant difference between bicalutamide 150 mg and castration in mean percent change from baseline in FFM (-1.56% and 3.86%, respectively, at week 96, p = 0.31), although there was a trend for greater progressive loss over time with castration. Mean changes in lipid parameters were small and similar in the 2 groups. CONCLUSIONS: Bicalutamide 150 mg monotherapy may offer an important advantage compared to castration in terms of bone loss and body composition for patients who require long-term androgen deprivation for localized or locally advanced prostate cancer. PMID- 15126802 TI - Racial differences in prostate androgen levels in men with clinically localized prostate cancer. AB - PURPOSE: We sought to determine whether there are racial differences in androgenic stimulation within the prostate tissue microenvironment. MATERIALS AND METHODS: Steroid hormones were extracted from snap frozen tissue obtained intraoperatively from radical prostatectomy specimens of 36 black and 59 white Americans. Testosterone, dihydrotestosterone (DHT), androstenedione (ASD), dehydroepiandrosterone, dehydroepiandrosterone sulfate, sex hormone-binding globulin (SHBG) and prostate specific antigen (PSA) were measured using radioimmunoassay. The Wilcoxon 2 group test was performed to compare clinical characteristics and tissue steroid levels between white and black Americans. Nonparametric rank ANOVA was used to consider race and other clinical factors in a multivariable way. RESULTS: Black and white American men were similar with respect to serum PSA, and pathological grade and stage. However, black men were younger (p = 0.01) and had a significantly higher body mass index (p = 0.02). Black and white men had similar testosterone and DHT. However, black men had higher ASD (p = 0.006) and SHBG (p = 0.009). Racial differences in ASD (p = 0.015) and SHBG (p = 0.008) persisted after controlling for age, body mass index, PSA, and pathological Gleason sum and stage. CONCLUSIONS: Tissue levels of testosterone and DHT did not differ by race. However, black men had higher tissue ASD and SHBG than white men. Higher tissue ASD did not result in a greater conversion of ASD to testosterone in the prostate of black men. Higher tissue SHBG may activate the androgen receptor through cyclic adenosine monophosphate dependent pathways. PMID- 15126803 TI - Association of tobacco use with hormone refractory disease and survival of patients with prostate cancer. AB - PURPOSE: We identified time to the development of hormone refractory prostate cancer (HRPC) and survival in men with advanced prostate cancer and examined any association with cigarette smoking history. MATERIALS AND METHODS: The study design was retrospective. A complete tobacco history was obtained in 222 patients with advanced prostate cancer. HRPC was diagnosed in 133 of these 222 patients and death occurred in 77. Standard statistical methods and software were used to analyze these data. RESULTS: Median time to HRPC was 11, 23 and 35 months in patients with a current, former or never smoked tobacco history (p = 0.00001). Median overall survival time on androgen ablative therapy was 38, 47 and 60 months in patients with a current, former or never smoked tobacco history, respectively (p = 0.00001). CONCLUSIONS: Tobacco use independently correlates with time to HRPC and survival in a dose dependent manner in patients with advanced prostate cancer on androgen deprivation therapy. PMID- 15126804 TI - Changing patterns in competing causes of death in men with prostate cancer: a population based study. AB - PURPOSE: We examined trends in hospitalization and death in men with prostate cancer to determine whether outcomes have changed with time in men diagnosed and treated for this disorder. MATERIALS AND METHODS: A population based cohort study of 180973 patients with prostate cancer in the 1979 to 1996 Surveillance, Epidemiology and End Results cancer registry and 450448 admissions in the 1987 to 1996 Surveillance, Epidemiology and End Results-Medicare linked database were analyzed. ORs derived from logistic regression were used to assess time trends in mortality and hospitalization. Multinominal logistic regression was used to obtain the adjusted proportions of deaths due to various causes in different years. RESULTS: In men with prostate cancer the risk of death from cancer was 39.7% (OR = 0.61, 95% CI = 0.56 to 0.66), which was lower in 1995 to 1996 than in 1979 to 1980. Decreases in prostate cancer death were greater than those in cardiovascular disorders (OR = 0.85, 95% CI = 0.78 to 0.92) and evident even in men with nonlocalized disease. Overall nonprostate cancer causes of mortality increased (OR = 1.65, 95% CI = 1.52 to 1.79) and ultimately exceeded that due to prostate cancer. By 1995 to 1996 the proportion of prostate cancer deaths was similar to that of cardiovascular disorders (27.7% and 26.6%, respectively) and substantially less than that of all other sources combined (45.7%). Similar effects were observed for prostate cancer (OR = 0.40, 95% CI = 0.37 to 0.42) and nonprostate cancer (OR = 2.51, 95% CI = 2.36 to 2.68) hospitalizations. CONCLUSIONS: In men with prostate cancer decreases in prostate cancer hospitalization and mortality have been greater than those in competing diseases with time. Most deaths in patients with prostate cancer, including those with nonlocalized disease, are now due to nonprostate cancer causes. PMID- 15126805 TI - Evaluating the risk of epididymal injury during hydrocelectomy and spermatocelectomy. AB - PURPOSE: We evaluated the risk of epididymal injury in patients undergoing hydrocelectomy and spermatocelectomy, and determined risk factors that may increase the chance of epididymal injury. To our knowledge the incidences of epididymal injury during hydrocelectomy and spermatocelectomy have not previously been reported. MATERIALS AND METHODS: The pathology reports of all patients undergoing hydrocelectomy and spermatocelectomy at a single institution from 1990 to 2003 were retrospectively reviewed to determine if a portion of the epididymis was present in the pathology specimen. Patients with epididymis present then underwent a chart review to determine possible risk factors for epididymal injury. RESULTS: A total of 338 adults underwent unilateral or bilateral hydrocelectomy from 1990 to 2003. Another 111 patients underwent spermatocelectomy during this period. In 19 patients (5.62%) epididymal injuries were documented during hydrocelectomy and in 19 (17.12%) epididymal injuries were documented during spermatocelectomy. No specific risk factors could be identified. CONCLUSIONS: The risk of epididymal injury during hydrocelectomy and spermatocelectomy is significant. Patients must be informed of this risk since epididymal injury may lead to infertility. To our knowledge this is the first published report documenting the incidence and risk of epididymal injury during hydrocelectomy or spermatocelectomy. PMID- 15126806 TI - Conversion from colonic or ileal conduit to continent cutaneous urinary diversion. AB - PURPOSE: After ileal or colonic conduit diversion some patients, particularly adolescents, desire conversion to a continent diversion to improve quality of life. We report our long-term results on conversion from conduit diversion to continent cutaneous diversion. MATERIALS AND METHODS: Between 1986 and 2001, 39 patients (mean age 24 years, range 6 to 49) underwent conversion from a colonic (21) or ileal conduit (18) to an ileocecal pouch (Mainz pouch I) with a mean followup of 102 months (range 18 to 192). Conversion was performed after a mean of 11 years (range 1 to 36) of conduit urinary diversion by incorporating the preexisting colonic/ ileal conduit and the ileocecal pouch. RESULTS: A total of 21 patients (54%) experienced complications requiring surgical intervention including stoma stenosis (13%), pouch calculi (31%) and ureteral stenosis (6 of 75 renoureteral units, 8%). Continence was achieved in 95% of patients. Defecation was unchanged in 72% of patients without treatment. In 21% fecal frequency was medically controlled (cholestyramine, loperamide) and 8% of patients had fecal frequency. During followup early substitution of alkali was performed and in 19 patients (49%) the venous base excess was less than -2.5 mmol/l to prevent hyperchloremia and acidosis. CONCLUSIONS: The inclusion of a preexisting colonic or ileal conduit decreases resection length of bowel for continent cutaneous diversion. Acceptable complication rates, stable renal function and satisfaction of patient expectations support conversion from a conduit into a Mainz pouch I as a safe and viable option in the long run. PMID- 15126807 TI - Modified ileal neobladder for continent urinary diversion: functional results after 9 years of experience. AB - PURPOSE: We report the continence status and urodynamic findings in 59 patients who underwent radical cystectomy and orthotopic bladder reconstruction using a modified S-ileal neobladder technique between January 1993 and July 2002. MATERIALS AND METHODS: Continence status was assessed using patient interview, frequency-volume charts and the need for protective devices. Reservoir sensation, compliance, capacity and activity were assessed by enterocystometry. Post-void residual urine volume greater than 100 ml was the indication for clean intermittent self-catheterization. RESULTS: Daytime continence was reported by 95.5% and 100% of our patients at 1 and 5 years, respectively. Overall, 77.7% of patients reported nighttime continence at 1 year and 96.5% at 5 years. The need for a protective device decreased with time and most of the patients would have undergone the operation again. Enterocystometric capacity and maximum reservoir pressure remained remarkably stable at 391.6 versus 440 ml, and 30 versus 20 cm H2O, 6 months and 5 years after surgery, respectively. Nevertheless, median post void residual urine volume increased from 35 ml at 6 months to 55 ml at 5 years with an increase in prevalence of patients requiring intermittent self catheterization due to post-void residual urine greater than 100 ml from 1.8% at 6 months to 10% at 5 years. CONCLUSIONS: Orthotopic bladder substitution with the modified S-ileal neobladder technique has an excellent functional outcome over time, resulting in high daytime and nighttime continence levels as well as high acceptability rates from our patients. PMID- 15126808 TI - Urinary diversion. PMID- 15126809 TI - Diagnosis of ureteral obstruction in patients with compromised renal function: the role of noninvasive imaging modalities. AB - We compared the role of noncontrast computerized tomography (NCCT), magnetic resonance urography (MRU), and combined abdominal radiography (KUB) and ultrasonography (US) in the diagnosis of the cause of ureteral obstruction in patients with compromised renal function. MATERIALS AND METHODS: The study included 149 patients, of whom 110 had bilateral obstruction and 39 had obstruction of a solitary kidney. Therefore, the total number of renal units was 259. All patients had renal impairment with serum creatinine greater than 2.5 mg/dl. Besides conventional KUB and US all patients underwent NCCT and MRU. The gold standard for diagnosis of the cause of obstruction included retrograde or antegrade ureterogram, ureteroscopy and/or open surgery. The sensitivity, specificity and overall accuracy of NCCT, MRU, and combined KUB and US in the diagnosis of ureteral obstruction were calculated in comparison with the gold standard. RESULTS: The definitive cause of ureteral obstruction was calculous in 146 and noncalculous in 113 renal units, including ureteral stricture in 65, bladder or ureter in 43, extraurinary collection in 3 and retroperitoneal fibrosis in 2. The site of stone impaction was identified by NCCT in all 146 renal units (100% sensitivity), by MRU in 101 (69.2% sensitivity), and by combined KUB and US in 115 (78.7% sensitivity) with a difference of significant value in favor of NCCT (p <0.001). Ureteral strictures were identified by NCCT in 18 of the 65 cases (28%) and by MRU in 54 of 65 (83%). Bladder and ureteral tumors causing ureteral obstruction could be diagnosed in approximately half of the patients by NCCT (22 of 43) and in all except 1 by MRU (42 of 43). NCCT and MRU could identify all extraurinary causes of obstruction. Overall of the 113 kidneys with noncalculous obstruction the cause could be identified by MRU in 101 (89% sensitivity), by NCCT in 45 (40% sensitivity), and by combined KUB and US in only 20 (18% sensitivity) with a difference of significant value in favor of MRU (p <0.001). CONCLUSIONS: In patients with renal impairment due to ureteral obstruction NCCT has superior diagnostic accuracy for detecting calculous causes of obstruction but MRU is superior for identifying noncalculous lesions. PMID- 15126810 TI - Patient experience with a urodynamic study: a prospective study in 208 patients. AB - PURPOSE: We assessed several emotional variables, including anxiety, pain, shame and morbidity, in patients who underwent a urodynamic study. MATERIALS AND METHODS: A total of 208 patients of 20 to 81 years old (mean age 54.3) were included in the study. Of the patients 71 (34.1%) were male and 137 (65.9%) were female. Prior to and immediately after the procedure each patient completed a self-administered questionnaire. Answers were given on a visual analog scale. All patients received an antibiotic for 3 days after the investigation and urine was collected for culture 7 days after the investigation. RESULTS: Patient initial anxiety did not significantly depend on demographics. The mean degree of pain +/- SEM experienced by patients was higher in males than in females (3.1+/- 0.2 vs 2.4 +/- 0.2, p = 0.012). Pain scores correlated with pre-procedure anxiety scores but the relationship was weak ([pain] = 0.283[anxiety] + 1.766, r = 0.157, p = 0.024). On the other hand, female patients showed a significantly higher mean level of shame than males (3.6 +/- 0.3 vs 1.8 +/- 0.3, p <0.001). In general patients cooperated extremely well during the examination. By the stepwise method only bother was associated with tolerance ([tolerance] = 0.182[bother] + 2.753, r = 0.275, p <0.001). Seven patients (3.4%) had significant bacteriuria and received antibiotic treatment. CONCLUSIONS: The study demonstrates that urodynamic studies are well tolerated by male and female patients in all age groups. However, emotional support depending on gender prior to the urodynamic procedure might enhance the level of patient cooperation. PMID- 15126812 TI - Alfuzosin once daily facilitates return to voiding in patients in acute urinary retention. AB - PURPOSE: We confirmed the beneficial effect of the alpha1-blocker alfuzosin for the acute management of acute urinary retention (AUR) related to benign prostate hyperplasia (BPH), and further identified factors influencing the success of a trial without catheter (TWOC). MATERIALS AND METHODS: A total of 360 patients presenting with a first episode of spontaneous AUR related to BPH underwent emergency catheterization and were then randomly and blindly assigned to receive 10 mg alfuzosin once daily or placebo at a ratio of 2:1 for 3 days. The primary efficacy criterion of this large study was the rate of successful TWOC within 24 hours after catheter removal. The influence of factors such as age, urine retention volume, fluid consumption, constipation and urinary tract infection on TWOC outcome was also assessed. RESULTS: Successful TWOC was recorded in 61.9% of the 236 patients treated with alfuzosin vs 47.9% of the 121 receiving placebo (p = 0.012). Elderly patients (65 years or older) and patients with a drained volume of 1000 ml or greater had significantly greater chances of TWOC failure (success vs failure OR 0.309, 95% CI 1.182 to 0.514 and OR 0.361, 95% CI 0.225 to 0.571, respectively). Nevertheless, even in the presence of these 2 factors 10 mg alfuzosin once daily almost doubled the likelihood of successful TWOC (OR 1.98, 95% CI 1,226 to 3,217). Alfuzosin (10 mg) once daily was well tolerated. CONCLUSIONS: Alfuzosin (10 mg) once daily significantly improved the rate of successful TWOC in patients with AUR related to BPH, even in elderly patients and those with a large drained volume who were at increased risk for TWOC failure. This should contribute to decrease the morbidity and mortality associated with emergency surgery and avoid the discomfort and potential morbidity associated with an in situ catheter. PMID- 15126811 TI - Trospium chloride improves overactive bladder symptoms: a multicenter phase III trial. AB - PURPOSE: Trospium chloride is an anticholinergic agent with predominantly peripheral nonselective antimuscarinic activity lacking central nervous system effects. It has no known drug-drug interactions, an advantage for patients taking many medications. Because these qualities may provide added benefit when treating patients with symptoms associated with overactive bladder (OAB) and urge incontinence, we studied the effectiveness of trospium in treating these conditions. MATERIALS AND METHODS: Patients with OAB with urge incontinence were randomized 1:1 to 20 mg trospium twice daily or placebo in this 12-week, multicenter, parallel, double-blind, placebo controlled trial. Dual primary end points were change in average number of toilet voids and change in urge incontinent episodes per 24 hours. Secondary efficacy variables were change in average of volume per void, voiding urge severity, urinations during day and night, time to onset of action and change in Incontinence Impact Questionnaire. RESULTS: A total of 523 patients were entered at 51 sites. Trospium significantly decreased average frequency of toilet voids and urge incontinent episodes compared to placebo. It significantly increased average volume per void, and decreased average urge severity and daytime frequency. All effects occurred by week 1 and all were sustained throughout the study. Nocturnal frequency decreased significantly by week 4 and Incontinence Impact Questionnaire scores improved at week 12. Trospium was well tolerated. CONCLUSIONS: Trospium was found to have sustained effectiveness beginning at the end of week 1 in decreasing the number of voids, urge incontinent episodes, total daily micturitions and urge severity, and in increasing volume per void. It also improved symptoms of OAB and quality of life. PMID- 15126813 TI - Validation of a clinical algorithm to diagnose stress urinary incontinence for large studies. AB - PURPOSE: We assessed the accuracy with which a clinical diagnostic algorithm for stress urinary incontinence (SUI) based on symptoms and signs without including urodynamics predicted the observation of urodynamic SUI and the condition of SUI. Such an algorithm would have applicability to ensure that the majority of women entering large SUI clinical trials would have urodynamic SUI and/or the condition of SUI without performing urodynamic testing in all subjects. MATERIALS AND METHODS: A total of 1455 women with urinary incontinence at least 3 months in duration were enrolled in 3 randomized clinical trials (1 phase 2 and 2 phase 3 trials) of duloxetine vs placebo for the treatment of SUI in Europe and North America. Subjects were identified as having SUI based on a clinical algorithm that required a predominant symptom of SUI with a weekly incontinence episode frequency of 4 or greater (phase 2 study), or 7 or greater (phase 3 studies), absent predominant urge symptoms, normal diurnal and nocturnal frequency, a bladder capacity of 400 ml or greater, and a positive cough stress (sign of SUI) and stress pad test. Multichannel urodynamic studies were performed in a subset of 184 women at 23 study sites. Of these urodynamic tracings 173 (94%) were of adequate quality to make an assessment of the filling phase and assign a diagnosis of urodynamic SUI, detrusor overactivity or a normal filling phase. Two sided 95% exact CIs for the proportions were calculated. RESULTS: The clinical algorithm had a positive predictive value of 90.2% for urodynamic SUI with or without detrusor overactivity and 76.9% for urodynamic SUI only (pure urodynamic SUI). The positive predictive value for the condition of pure SUI was 85.0%, while for the condition of SUI in pure and mixed forms the positive predictive value was 98.3%. Patient age, previous continence surgery or the severity of incontinence did not influence algorithm accuracy. CONCLUSIONS: The algorithm is suitably feasible and sufficiently predictive to be used in large clinical trials designed to evaluate conservative treatment for women with SUI. It ensures that the overwhelming majority of the study population would have urodynamic SUI and the condition of SUI. PMID- 15126814 TI - Granulomatous prostatitis linked to HLA-DRB1*1501. AB - PURPOSE: Granulomatous prostatitis is characterized by a pattern of granulomatous inflammation in the prostate. In most cases the etiology is unknown. Based on the hypothesis that granulomatous prostatitis may be an autoimmune disease we performed intermediate and selective high resolution typing of HLA-DR in a group of patients with the disease and compared the frequency of class II HLA phenotypes to that in a control group of volunteer marrow donors in the military. MATERIALS AND METHODS: Histological records from 1 institution from 1990 to 2000 revealed 12 patients with diffuse granulomatous prostatitis. Three patients were dead and 1 refused blood drawing. Peripheral blood from the remaining 8 patients was typed along with blood from an additional 3 identified at the practice of one of us from 1999 through 2002. All slides were reviewed by 1 pathologist. Intermediate resolution typing of HLA-A, B and DR was performed by polymerase chain reaction-sequence specific oligonucleotide probe. High resolution, allele specific identification of HLA DR15 was performed if patients were DR15 positive by intermediate resolution typing. RESULTS: There were 3 black and 8 white individuals identified with diffuse nonspecific granulomatous prostatitis. Six of 8 white patients (75%) were HLA-DR15 by intermediate resolution typing. One of the 3 black American patients (33%) was HLA-DR15. In the control group 127 of 451 white (28.2%) and 23 of 89 black (25.8%) volunteer marrow donors were HLA-DR15. The case-control comparison of white patients was significantly different (Fisher's exact test p = 0.0086). There were no statistically significant differences between case-control comparisons for any other HLA-DR phenotype. High resolution DR15 typing showed that the white patients were HLA-DRB1*1501 and the black patient was HLA-DRB1*1503. CONCLUSIONS: The data suggest an association between HLA-DRB1*1501 and granulomatous prostatitis. HLA-DR15 is strongly associated with other autoimmune diseases, notably multiple sclerosis. The data are consistent with an autoimmune etiology for nonspecific granulomatous prostatitis. PMID- 15126815 TI - Association of prostatic inflammation with down-regulation of macrophage inhibitory cytokine-1 gene in symptomatic benign prostatic hyperplasia. AB - PURPOSE: Our previous DNA microarray analyses revealed that the macrophage inhibitory cytokine-1 (MIC-1) gene was significantly down-regulated in symptomatic benign prostatic hyperplasia (BPH) samples. We compared the histopathological features of inflammatory changes in prostate adenoma tissues from individuals with symptomatic and asymptomatic (histological only) BPH using MIC-1 mRNA levels. MATERIALS AND METHODS: Prostate adenoma tissues were obtained from 25 patients who underwent transurethral prostatectomy to relieve lower urinary tract symptoms due to BPH and 6 patients with bladder cancer who underwent cystoprostatectomy due to bladder cancer. Inflammatory changes in the prostate were examined histopathologically and immunohistochemically, and classified according to the consensus classification system of the Chronic Prostatitis Collaborative Research Network and International Prostatitis Collaborative Network advocated in 2001 with some modification. MIC-1 mRNA was assessed quantitatively by real-time reverse transcriptase-polymerase chain reaction. RESULTS: MIC-1 gene down-regulation was observed in 16 of 25 symptomatic BPH samples (64.0%), whereas MIC-1 mRNA was high in 6 of 6 prostate adenoma samples from patients with bladder cancer (p = 0.0006). MIC-1 gene down regulation correlated with the grade of glandular/ periglandular inflammatory changes with statistical significance (p = 0.0387). MIC-1 gene down-regulation was found in only 1 of 7 BPH samples with a glandular predominant pattern, whereas it was found in 15 of 24 BPH samples with a mixed type or stromal predominant pattern (p = 0.0373). CONCLUSIONS: Gland destruction by inflammatory infiltrates, followed by replacement of the stromal component in symptomatic BPH may be induced by the down-regulation of the MIC-1 gene. PMID- 15126816 TI - Transurethral needle ablation versus transurethral resection of the prostate for the treatment of symptomatic benign prostatic hyperplasia: 5-year results of a prospective, randomized, multicenter clinical trial. AB - PURPOSE: We report the 5-year efficacy and safety of transurethral needle ablation of the prostate (TUNA) compared to transurethral resection of the prostate (TURP) for the treatment of lower urinary tract symptoms (LUTS) secondary to benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: A total of 121 men 50 years or older with LUTS secondary to BPH a minimum of 3 months in duration were enrolled in this prospective, randomized clinical trial at 7 medical centers across the United States. Of the participants 65 (54%) were randomly selected to receive TUNA and 56 (46%) were selected to receive TURP. International Prostate Symptom Score, quality of life, peak urinary flow rate, post-void residual urinary volume, and prostate size and configuration were evaluated before the procedure and then annually for 5 years after the procedure. Adverse events were also recorded throughout the study. RESULTS: Improvement from baseline for TUNA and TURP retained statistical significance at each interval for International Prostate Symptom Score, quality of life and peak flow rate. Post void residual volume was statistically significant at all time points for TURP and at year 5 for TUNA. The TURP group reported 41% retrograde ejaculation, while the TUNA group reported none. The incident of erectile dysfunction, incontinence and stricture formation was also greater in TURP than in TUNA cases with significantly fewer adverse events for TUNA than for TURP. CONCLUSIONS: The results of this study demonstrate stable treatment outcomes after 5 years of followup and suggest that TUNA is an attractive treatment option for men with LUTS due to BPH. PMID- 15126817 TI - The prevalence of hypertension, hyperlipidemia, diabetes mellitus and depression in men with erectile dysfunction. AB - PURPOSE: We quantified the prevalence of diagnosed hypertension, hyperlipidemia, diabetes mellitus and depression in male health plan members with erectile dysfunction (ED). MATERIALS AND METHODS: We used a nationally representative managed care claims database that covered 51 health plans with 28 million lives for 1995 through 2002. Based on 272325 identified patients with ED population and age specific prevalence rates were calculated for the same period. RESULTS: The crude population prevalence rates were 41.6% for hypertension, 42.4% for hyperlipidemia, 20.2% for diabetes mellitus, 11.1% for depression, 23.9% for hypertension and hyperlipidemia, 12.8% for hypertension and diabetes mellitus, and 11.5% for hyperlipidemia and depression. The crude age specific prevalence rates varied across age groups significantly for hypertension (4.5% to 68.4%), hyperlipidemia (3.9% to 52.3%), and diabetes mellitus (2.8% to 28.7%), and significantly less for depression (5.8% to 15.0%). Region adjusted population prevalence rates were 41.2% for hypertension, 41.8% for hyperlipidemia, 19.7% for diabetes mellitus and 11.9% for depression. Only 87163 patients with ED (32%) had no comorbid diagnosis of hypertension, hyperlipidemia, diabetes mellitus or depression. CONCLUSION: Hypertension, hyperlipidemia, diabetes mellitus and depression were prevalent in patients with ED. This evidence supported the proposition that ED shares common risk factors with these 4 concurrent conditions. Therefore, as a pathophysiological event, ED could be viewed as a potential observable marker for these concurrent diseases. This finding suggests that clinicians could include ED in the assessment profile of these concurrent conditions for earlier detection and treatment. PMID- 15126818 TI - Combined penile plication surgery and insertion of penile prosthesis for severe penile curvature and erectile dysfunction. AB - PURPOSE: For persistent penile curvature after corporeal dilation and prosthesis placement other maneuvers, such as plaque incision with or without grafting, or penile modeling, must be used to complete penile straightening. However, each of these techniques is associated with increased morbidity, which includes higher rates of infection and urethral injury. We describe a new technique for correcting severe penile curvature and erectile dysfunction using combined penile plication and placement of a 3 piece inflatable penile prosthesis. MATERIALS AND METHODS: Between February 2000 and June 2003, 5 patients with erectile dysfunction and severe curvature for which other treatment modalities had failed were treated with combined inflatable penile prosthesis and additional plication to correct the severe malformation. Detailed medical and sexual history was obtained as well as preoperative duplex ultrasound prior to surgery. RESULTS: The age range of our patient population was 22 to 55 years. In each case penile prosthesis placement was insufficient to correct the malformation and the addition of multiple plication sutures was required. The malformation in all patients was corrected with this technique. To date no patients have reported any complications with followup (range 3 to 36 months). CONCLUSIONS: In cases of severe penile deformity and curvature with erectile dysfunction the combination of penile plication and inflatable penile prosthesis placement is a method of repair that is well tolerated. PMID- 15126819 TI - Subjective and objective analysis of the prevalence of Peyronie's disease in a population of men presenting for prostate cancer screening. AB - PURPOSE: In this study we defined the prevalence of Peyronie's disease in a cohort of men being screened for prostate cancer in the United States. The association between Peyronie's disease, and medical comorbidities and patient self-reported erectile dysfunction was also defined. MATERIALS AND METHODS: A total of 534 men presenting to 1 of 3 prostate cancer screening centers provided a complete medical history, underwent physical examination performed in all by a urologist and completed the Sexual Health Inventory for Men (SHIM) questionnaire. This population was chosen because of the access to a large number of subjects combined with the fact that subjects were not presenting with a specific urological complaint. The diagnosis of Peyronie's disease was based on a palpable penile plaque. Data were assessed using univariate analysis, multivariate logistic regression and Pearson chi-square analysis. RESULTS: A total of 48 patients were found to have a palpable penile plaque on physical examination for a prevalence rate of 8.9%. The mean age of men with Peyronie's disease was 68.2 years compared to a mean of 61.8 years in men without Peyronie's disease (p <0.0001). On univariate analysis hypertension (p = 0.02) and diabetes (p = 0.007) were present with significantly increased frequency in patients with Peyronie's disease. Patients diagnosed with Peyronie's disease had significantly lower values for responses to each of the 5 questions on the SHIM survey. An increase in age and decrease in total SHIM score remained significantly associated with a greater probability of Peyronie's disease on multivariate analysis. CONCLUSIONS: We found the prevalence of Peyronie's disease to be greater than in most previously reported series. We also noted a significant association between Peyronie's disease aging, hypertension, diabetes and self-reported erectile dysfunction. PMID- 15126820 TI - An artificial neural network for predicting the presence of spermatozoa in the testes of men with nonobstructive azoospermia. AB - PURPOSE: We developed an artificial neural network (ANN) for predicting spermatozoa prior to testicular biopsy in men with nonobstructive azoospermia. The performance of this ANN was compared to that of the standard logistic regression (LR) model. MATERIALS AND METHODS: Data were retrospectively collected from the physical examination and laboratory records of 303 patients who presented with infertility due to nonobstructive azoospermia. Input factors were patient age, duration of infertility, serum follicle-stimulating hormone, luteinizing hormone, total testosterone and prolactin, and left and right testicular volume. The ANN and LR models were constructed based on data on cases in which spermatozoa were and were not detected on testicular sperm extraction. The ANN was trained and validated with the data in the training (200 cases) and validation (30 cases) sets, and the model was then used to predict findings in the test set (73 cases). The LR model was constructed using the same data on the 230 training and validation cases. The same 73 patients served as the test set. RESULTS: The sensitivity of the ANN model was significantly higher than that of the LR model (68% vs 28%, p < 0.0001). The neural network correctly predicted the outcome in 59 of the 73 test set patients (80.8%), whereas LR correctly predicted the outcome in 48 (65.7%, p = 0.07). CONCLUSIONS: This ANN model, which is based on age, duration of infertility, serum hormone levels and testicular volumes, has clinically acceptable sensitivity. It may be of value for predicting spermatozoa in men with nonobstructive azoospermia. PMID- 15126821 TI - Role of androgens in erectile function. AB - PURPOSE: Erectile response in mammals is centrally and peripherally regulated by androgens. Severe hypogonadism in men usually results in loss of libido and potency. The present studies were designed to evaluate the possible influence of circulating androgens in the regulation of sexual function. MATERIALS AND METHODS: A total of 15 men with severe hypogonadism (testosterone less than 2.0 ng/ml) were recruited. The control group consisted of 20 patients with psychogenic erectile dysfunction. All subjects underwent nocturnal penile tumescence (NPT) and rigidity monitoring during 2 consecutive nights, evaluation of cavernous artery flow using penile color duplex ultrasound (P-CDU), and real time visually stimulated erection evaluation at baseline and after administration of 50 mg sildenafil or 3 mg apomorphine. NPT, P-CDU and visually stimulated erection were evaluated after 6 months of therapy with a 5 mg daily testosterone patch. RESULTS: NPT analysis in subjects with severe hypogonadism showed a significant decrease in sleep related erections. P-CDU showed a partial erectile response with an alteration of the parameters analyzed, and the visually stimulated erections test was pathological. Administration of 3 mg apomorphine and 50 mg sildenafil has no influence on erectile function. Testosterone treatment for 6 months induced normalization of NPT and P-CDU parameters, and of visually stimulated erection effects with the restoration of a normal response to pharmacological stimulation with apomorphine or sildenafil. CONCLUSIONS: Our results show that testosterone has a key role in the central and peripheral modulation of erectile function even if the accurate testosterone plasma level threshold that may influence these processes remains to be established. PMID- 15126822 TI - 2003 American Urological Association Gallup survey: physician practice patterns, cryosurgery/brachytherapy, male infertility, female urology and insurance/professional liability. AB - PURPOSE: The Health Policy Survey and Research Committee of the American Urological Association and the Gallup Organization have performed 10 surveys of American urologists since 1992 for the purpose of assessing demographics and practice patterns. The results of the 2003 survey are presented. MATERIALS AND METHODS: A random sample of 510 urologists who have completed urological residencies was interviewed by telephone in February 2003. Major content areas were physician practice patterns, cryosurgery/brachytherapy, male infertility, female urology and insurance/ professional liability. RESULTS: Urologists are older, staying in practice longer and planning to retire later than ever before. Urologists are also seeing more patients in the office every year. Most urologists treat male infertility and female voiding dysfunction. CONCLUSIONS: American urologists are older, working harder and planning on continuing to do so for longer than in any year sampled in the last decade. PMID- 15126823 TI - Continuous needle vented foley catheter suction for urinary leak after radical prostatectomy. AB - PURPOSE: Prolonged urinary leakage from the urethrovesical anastomotic site after radical retropubic prostatectomy is a relatively uncommon complication. After the failure of other conservative measures we successfully overcame this problem using a Foley catheter needle vented suction technique. MATERIALS AND METHODS: Anastomotic leakage was noted after retropubic prostatectomy. After the failure of conservative techniques, including catheter traction, passive drainage and active catheter suction, needle vented suction was used. An 18 gauge needle was inserted into the plastic suction tubing attached to the Foley catheter to prevent suction occlusion of the eye of the Foley catheter by the bladder mucosa. RESULTS: Needle vented suction successfully treated urinary leakage after radical retropubic prostatectomy in 2 patients. CONCLUSIONS: In cases of prolonged anastomotic urinary leakage after radical retropubic prostatectomy needle vented suction can be considered an alternative conservative approach to treatment. PMID- 15126824 TI - A surgical method to preserve testicular function and restore cosmetic appearance in hypogonadal men. AB - PURPOSE: Maintenance of spermatogenesis in hypogonadotropic hypogonadal patients is possible and is an objective to be actively pursued. Gonadotropin releasing hormone and gonadotropin/ testosterone therapy can develop and maintain sex characteristics and spermatogenesis such that even small gonads should be preserved. An empty scrotum, like any other congenital or acquired condition compromising the size of the genitalia, can produce serious psychosexual problems and have an extremely negative effect on normal social life. MATERIALS AND METHODS: In 2 adolescents affected by Kallmann's syndrome with testes of infantile volume 1 gonad was transferred to the contralateral hemiscrotum and replaced by an adult testicular prosthesis. RESULTS: There were no postoperative complications. The cosmetic results were considered satisfactory by the patients and their parents. CONCLUSIONS: When possible and when there is no risk of physiological repercussions, every effort should be made to normalize the appearance of the genital area for strong psychological reasons. PMID- 15126825 TI - The epididymal slide technique: a tension-free anastomosis in a vasoepididymostomy. AB - PURPOSE: Vasoepididymostomy is a technically demanding procedure that requires excellent positioning of the abdominal vas deferens at the epididymal tubule as well as a tension-free anastomosis. We describe a novel technique of securing the vas deferens to the testicle that allows manipulation of the epididymis and ensures a tension-free anastomosis. MATERIALS AND METHODS: After the vas deferens and epididymis are isolated in the usual fashion the vas is brought to the lateral portion of the testicle. A stay stitch is placed on the abdominal vas. This stay stitch is then placed through the tunica albuginea immediately posterior to the corpus or cauda epididymis. This allows the epididymis to slide in cephalad and caudad fashion prior to vasoepididymostomy and allows a tension free anastomosis. RESULTS: We have used this technique routinely for vasoepididymostomy without any associated complications and have found it to be technically feasible in our last 50 cases. CONCLUSIONS: Although it is simple, the epididymal slide technique allows minimal to no distortion of the epididymal tubule, secure and safe fixation of the vas deferens, flexibility in choosing an appropriate epididymal tubule and tailoring of tension at the conclusion of the procedure. This technique is now used routinely at our center to allow a tension free anastomosis during vasoepididymostomy and it has been technically feasible in all of our last 50 cases. PMID- 15126826 TI - Medical management of renal actinomycosis. PMID- 15126827 TI - Undifferentiated sex cord-stromal testis tumor. PMID- 15126828 TI - Synovial sarcoma of the prostate. PMID- 15126829 TI - Long-term followup of metastatic prostate cancer. PMID- 15126830 TI - Appendiceal substitution of the ureter in retroperitoneal fibrosis. PMID- 15126831 TI - Spontaneous bladder wall abscess. PMID- 15126832 TI - Intracavernous prostaglandin E1 infusion in diabetes with associated ischemic necrosis of the glans penis. PMID- 15126833 TI - Chronic pelvic actinomycosis following a bone anchored sling procedure. PMID- 15126834 TI - Amyloidosis of the seminal vesicle and hematospermia. PMID- 15126835 TI - Re: partial nephrectomy for unilateral wilms tumor: results of study SIOP 93 01/GPOH. PMID- 15126837 TI - Re: bladder cancer facts: accuracy of information on the internet. PMID- 15126839 TI - Re: tumor markers in the diagnosis of primary bladder cancer. A systematic review. PMID- 15126840 TI - Re: complexed prostate specific antigen improves specificity for prostate cancer detection: results of a prospective multicenter clinical trial. PMID- 15126841 TI - Re: contrast enhanced color Doppler endorectal sonography of the prostate: efficiency for detecting peripheral zone tumors and role for biopsy procedure. PMID- 15126843 TI - Re: reconstruction of the hypospadiac hooded prepuce. PMID- 15126845 TI - Re: air embolism from pneumopyelography. PMID- 15126846 TI - Re: 1-step removal of encrusted retained ureteral stents. PMID- 15126848 TI - Re: results of complete penile disassembly for epispadias repair in 42 patients. PMID- 15126850 TI - Re: an artificial somatic-central nervous system-autonomic reflex pathway for controllable micturition after spinal cord injury: preliminary results in 15 patients. PMID- 15126851 TI - Re: functional and neuroanatomical effects of vaginal distention and pudendal nerve crush in the female rat. PMID- 15126853 TI - Re: a new high frequency electrostimulation device to treat chronic prostatitis. PMID- 15126855 TI - Re: testicular sperm extraction with intracytoplasmic sperm injection is successful for the treatment of nonobstructive azoospermia associated with cryptorchidism. PMID- 15126856 TI - Re: tadalafil has no detrimental effect on human spermatogenesis or reproductive hormones. PMID- 15126859 TI - Bilateral robotic assisted laparoscopic heminephroureterectomy. PMID- 15126860 TI - Does bladder preservation (as a surgical principle) lead to retaining bladder function in bladder/prostate rhabdomyosarcoma? Results from intergroup rhabdomyosarcoma study iv. AB - PURPOSE: We determine patient and tumor characteristics, event-free and overall survival, methods of local control, rate of bladder preservation and proportion with normal bladder function for patients with localized bladder/prostate (BP) rhabdomyosarcoma (RMS) treated on the Fourth Intergroup Rhabdomyosarcoma Study (IRS IV). MATERIALS AND METHODS: We reviewed the records of 90 patients with nonmetastatic BP RMS enrolled on IRS IV for presenting characteristics, details of therapy and outcome. RESULTS: Of the 90 records 88 had sufficient information for review. Patient age distribution was less than 1 year for 7 patients, 1 to 9 years for 71 and 10 or greater years for 10. Tumors commonly arose in the bladder (70%), had favorable histology (embryonal or botryoid 80%), large (69% greater than 5 cm), unresectable (84% group III) and invasive (56% T2). Local therapy included radiation in 74 patients, and most patients underwent second-look operations after radiation. All patients received alkylating based chemotherapy. With a median followup of 6.1 years there have been 3 second malignancies, 1 toxic death and 18 relapses, for an event-free survival rate of 77%. Bladders were retained without relapse at last contact in 55 patients. Of those 55 patients 36 and of the entire group 40% had normal function determined by history. CONCLUSIONS: Of patients with nonmetastatic BP RMS on IRS IV 82% survived 6 years. Bladder function was preserved in 55% (36/66) of event-free survivors. Of all patients entered on study 40% (36 of 88) survive event-free with apparently normal functioning bladders. More precise long-term evaluation of bladder and sexual function will require application of better tools such as urodynamic studies and validated patient surveys. PMID- 15126861 TI - Tubularized incised plate hypospadias reoperation. AB - PURPOSE: We report our updated experience with tubularized incised plate hypospadias reoperations in a series of patients of whom the majority had undergone prior urethral plate incision. MATERIALS AND METHODS: Records of 31 consecutive patients undergoing tubularized incised plate reoperation were reviewed. The decision for this repair was based on a preserved urethral plate that appeared supple despite prior surgery. RESULTS: Of the 31 patients the mean number of prior operations was 1.1, including 18 (58%) who had undergone primary repairs that involved midline plate incision. Overall, 28 (90%) patients had a successful outcome with a functional neourethra and vertical slit meatus. Complications occurred in 7 (23%) patients, consisting mostly of fistulas. Among 27 cases in which dartos was used as a barrier layer fistulas occurred in 1 (6%) of 18 when a ventral flap was placed over the neourethra versus 3 (33%) of 9 when adjacent tissues alongside the neourethra were approximated in the midline (p = 0.055). The rate of complications was not affected by history of urethral plate incision. In 3 patients partial or complete glans dehiscence or a large fistula occurred, and 2 subsequently required staged buccal graft urethroplasty. CONCLUSIONS: Tubularized incised plate reoperation results in a functional neourethra with a vertical slit meatus when the plate has been preserved and appears supple after prior surgery. Fistulas are less likely when a flap is interposed between the neourethra and skin. Complications are low despite previous urethral plate incision if there is no apparent scarring of the plate. An alternative technique for reoperative urethroplasty should be considered if the urethral plate has been excised or is grossly scarred. PMID- 15126862 TI - Octyl-2-cyanoacrylate as a routine dressing after open pediatric urological procedures. AB - PURPOSE: Octyl-2-cyanoacrylate Dermabond, Ethicon, Inc., Sommerville, New Jersey is a synthetic tissue adhesive that has recently been used for skin closure in the treatment of minor lacerations. We assess its effectiveness as a sole dressing after open pediatric urological procedures. MATERIALS AND METHODS: Between February and August 2003 we prospectively evaluated patients undergoing extragenital open and laparoscopic pediatric urological procedures at our institution. All open incisions were closed in layers using a final layer of self absorbing subcuticular stitches for the skin before applying Dermabond only at the skin level. In laparoscopic cases Dermabond alone was applied to port sites (3 mm or less) and instrument sites without any sutures beneath the skin. No adjuvant occlusive dressings were applied to any of these wounds. All patients were allowed to bathe and return to activity immediately postoperatively. RESULTS: During the study period 146 patients with 200 incisions were identified (open 146, laparoscopy 54). Of these children 103 (142 incisions) returned for followup during the period of study with only 1 complication identified. This 6 month-old child presented the evening of surgery with omental prolapse through the umbilical port (3 mm port) requiring urgent closure. In no case was there an appreciable healing problem on surgeon or parental examination and no wound infections occurred. CONCLUSIONS: Dermabond alone provides a simple coverage for a myriad of pediatric urological surgical wounds. Early bathing and return to activity do not appear to impact negatively on wound healing when a simple skin barrier is used in place of standard dressings. PMID- 15126863 TI - Long-term bladder dysfunction and renal function in boys with posterior urethral valves based on urodynamic findings. AB - PURPOSE: Posterior urethral valves are the most common cause of congenital obstructive uropathy leading to renal failure in childhood. We investigate the influence of bladder dysfunction on renal function impairment. MATERIALS AND METHODS: We retrospectively reviewed the records of 116 patients with posterior urethral valves. After valve ablation urodynamic studies were performed in all patients. The presence of vesicoureteral reflux (VUR), renal dysplasia, serum creatinine during followup as well as urodynamic abnormalities were recorded. Mean followup was 10.3 years after valve ablation. RESULTS: All patients underwent endoscopic valve ablation and urinary diversion was performed in 32. Renal dysplasia was found in 9 patients. Renal function impairment at the end of followup was present in 35 patients. Urodynamic studies showed poor compliance in 30 boys, detrusor overactivity in 44, and poor compliance and detrusor overactivity in 17. Bilateral VUR was found in 17 boys at the time of diagnosis. Urodynamic studies were normal in 23 (20%) patients, of whom 4 had renal failure. Univariate analysis showed that poor compliance and detrusor overactivity had a significant correlation to renal function impairment in addition to bilateral VUR and renal dysplasia. In a multivariate analysis bilateral VUR was an independent prognostic factor for poor prognosis. CONCLUSIONS: Bladder dysfunction may contribute to renal function impairment eventually but normal urodynamic findings do not preclude renal deterioration. It is likely that loss of compliance and detrusor overactivity would actually result in a valve bladder reaching its end point function. PMID- 15126864 TI - The modified sting procedure to correct vesicoureteral reflux: improved results with submucosal implantation within the intramural ureter. AB - PURPOSE: With the advent of tissue bulking agents, in particular dextranomer/hyaluronic acid copolymer (Dx/HA), for endoscopic implantation for vesicoureteral reflux (VUR), there has been a major shift in the surgical paradigm throughout Europe, and more recently, in the United States. We describe a modification of the technique used for implantation that has significantly improved our results. MATERIALS AND METHODS: Between October 2001 and October 2003, 285 children 7 months to 15 years old (mean age 4.6 years) underwent endoscopic implantation of Dx/HA for VUR at our institutions. A modified STING (subureteral transurethral injection) procedure (implantation submucosally within the intramural ureter) was introduced during the last year of the study. The average volume of injected material was measured for each ureter. Renal sonography was performed to determine if hydronephrosis was present. At 3 months flouroscopic voiding cystourethrograms were used to evaluate for the presence of VUR. A subset of 122 patients treated with STING (52) were compared to those treated with modified STING (70). RESULTS: A total of 459 ureters in 231 girls and 54 boys were treated (174 bilateral cases). Mean maximum grade per patient was 2.5/5. Mean injected volume was 0.9 cc ureter. There were 181 patients with at least 3 months of followup. After 1 treatment 76% (137 of 181) of cases were cured (grade 0 reflux), while 54% (24 of 44) of the failures were improved. The overall cure rate was 94% for grade I, 85% for grade II, 78% for grade III and 71% for grade IV reflux. The patients treated with STING had a mean age of 4.8 years, mean maximum reflux grade was 2.5 and success rate was 71% (37 of 52; 86% grade I, 89% grade II, 70% grade III and 63% grade IV reflux). The patients treated with a modified STING had a mean age of 5.5 years, mean maximum grade was 2.8 and a success rate was 89% (62 of 70; 100% grade I, 92% grade II, 91% grade III and 90% grade IV reflux). Ureteral success rates were significantly (p <0.01) greater for the modified STING (92%) vs the standard STING (79%). There were no cases of hydronephrosis at 3 months postoperatively. CONCLUSIONS: The majority of patients undergoing minimally invasive therapy for VUR with Dx/HA are cured after 1 treatment. The modified STING is our preferred method of implant injection for the correction of VUR and in our hands produces a resolution rate of 89% (92% of ureters). The technique optimizes ureteral coaptation, is easy to perform and is not associated with any significant short-term complications. Persistence of VUR in a minority of patients continues to be the only significant adverse effect of endoscopic implantation. PMID- 15126865 TI - Serum potassium and creatinine changes following unstented bilateral ureteral reimplantation in children. AB - PURPOSE: We assess the incidence of electrolyte and creatinine changes following unstented bilateral ureteral reimplantation and attempt to identify associated risk factors. MATERIALS AND METHODS: A total of 107 consecutive children with bilateral vesicoureteral reflux underwent bilateral unstented ureteral reimplantation. Study exclusion criteria were plication or tapering of any ureter, age less than 1 year and/or baseline serum creatinine greater than twice normal for age. Postoperatively serum electrolytes and creatinine were assessed by venous puncture until values normalized. The presence of nausea, vomiting, urinary retention and oliguria were recorded. RESULTS: Of the patients 46 females and 10 males 1.0 to 10.9 years old met the study criteria and had complete data available. Four patients (7.1%) had postoperative potassium greater than or equal to 5.0 mmol/l (range 5.0 to 5.3), including 3 (75%, p = 0.0238) who received potassium supplemented intravenous fluid postoperatively. Eight (14.3%) patients had postoperative creatinine greater than 1 mg/dl (range 1.3 to 2.3) and concurrent hyperkalemia with increased creatinine occurred in 2 (25%, p = 0.0295). Nausea and vomiting beyond postoperative day 1 were noted in 6 patients (75%, p = 0.0122). Neither oliguria nor urinary retention reached statistical significance in correlation with increased potassium and/or creatinine. However, urine retention approached statistical significance in patients with increased creatinine (p = 0.0747). No adverse effects from hyperkalemia were noted. CONCLUSIONS: Adverse effects from hyperkalemia following unstented bilateral ureteral reimplantation are uncommon. Potassium containing intravenous fluids should be avoided in the early postoperative period. Routine serum electrolyte determination may be helpful in patients undergoing unstented bilateral ureteral reimplantation when persistent nausea, emesis or urinary retention is present. PMID- 15126866 TI - Complications associated with clean intermittent catheterization in children with spina bifida. AB - PURPOSE: Clean intermittent catheterization (CIC) is commonly used in the management of voiding dysfunction associated with neurospinal dysraphism. We determined the incidence of genitourinary complications associated with this technique in a population of children with spina bifida. MATERIALS AND METHODS: The charts of all children younger than 13 years attending a multi-disciplinary spina bifida clinic between 1987 and 2002 were reviewed. Children in whom CIC had been performed for a minimum of 5 years were identified. Catheterization was performed with a polyvinyl chloride catheter and a water-based lubricant. All genitourinary complications that had occurred in this group were recorded. The caregiver of each patient was then contacted by telephone to confirm the accuracy of our data. RESULTS: There were 32 females and 27 males identified in whom CIC had been performed for a minimum of 5 years. Mean duration of CIC was 10.5 years (range 5 to 15). Of the patients 45 (76%) learned to perform CIC independently at a mean age of 8 years (range 4.7 to 15.3). Two complications were gross hematuria and a false passage in the bulbar urethra. Both complications occurred in males early in the course of CIC, and while being performed by a caregiver. Neither complication was associated with long-term sequelae. CONCLUSIONS: The incidence of genitourinary complications associated with CIC in children with spina bifida is low. We identified 2 complications during a period of 570 patient-years for an incidence of 3.5 complications/1000 patient-years (95% confidence interval -1.3, 8.3) of observation. PMID- 15126867 TI - Does mode of treatment affect the outcome of neonatal posterior urethral valves? AB - PURPOSE: A prospective study on the outcome of posterior urethral valves (PUV) was performed. The data analyzed were whether the modality of treatment (fulguration vs vesicostomy) affected renal function and somatic growth, and whether the presence of vesicoureteral reflux (VUR) and abnormal serum creatinine levels affected somatic growth. MATERIALS AND METHODS: A total of 45 consecutive neonates were diagnosed and treated for PUV with fulguration (24) or vesicostomy (21) between 1997 and 2003. Postoperative stable creatinine values, renal function and somatic growth were recorded. Well tempered renal scans using diethylenetetraminepentaacetic acid were performed during the first and second years of life at followup. Standard anthropometric techniques and statistical methods were used to compute distance statistics for body weight and crown-heel length at age intervals of 3 months for year 1 and 6 months for year 2. RESULTS: Of the patients 9 were lost to followup and 6 died in the first year of life due to renal failure. Preoperative and postoperative mean serum creatinine was 1.6 +/ 1.5 and 0.7 +/- 0.2 mg/dl, for the fulguration group and 1.7 +/- 1.5 and 0.9 +/- 0.7 mg/dl, respectively, for the vesicostomy group. In 10 patients renal function deteriorated or there was no improvement after treatment. With the advancement of age neonates with PUV showed normal increase in body weight and crown-heel length. Those with VUR were significantly shorter compared to neonates with no VUR at the end of year 2 of life (p <0.05). Patients with creatinine less than 1 mg/dl had significantly greater increase in body weight (at 3 and 6 months) as well as crown-heel length (at 3 and 9 months), respectively (p <0.05). Physical growth of the fulguration and vesicostomy groups remained substantially lower than that of their normal healthy counterparts. Newborns treated with vesicostomy did not have any breakthrough urinary tract infections. Although patients treated with vesicostomy were lighter and smaller initially (p <0.05), they were comparable to those treated with fulguration at the end of 2 years, thus demonstrating catch-up growth. CONCLUSIONS: Our prospective study demonstrated that transurethral fulguration and vesicostomy are equally effective for neonatal valves and achieve similar renal function. Both groups showed retarded growth compared to healthy counterparts. Somatic growth was delayed by serum creatinine greater than 1.0 mg/dl and the presence of VUR. Vesicostomy seemed to help neonates catch-up the growth deficit in the first 2 years of life. PMID- 15126868 TI - Structural evidence against hormonal therapy for cryptorchid testis: abnormal gubernacular attachment. AB - PURPOSE: Various factors are involved in testicular descent, and no single factor is elucidated as the cause of cryptorchidism. We prospectively observed the locations of gubernacular attachments in the cryptorchid testis. MATERIALS AND METHODS: From April 1996 to June 2002, 639 boys underwent surgical correction of cryptorchid testis. Those with complete records of testicular locations and gubernacular attachments were analyzed. RESULTS: A total of 639 boys underwent 732 surgical corrections of inguinal testis (425, 58%), high scrotal testis (165, 23%) and impalpable abdominal testis (142, 19%). Impalpable abdominal testes had gubernacular attachment to either the lateral or upper scrotum in 36 cases (25%) or around the inguinal ring in 83 (58%). The gubernaculum was attached to the bottom of the scrotum in 21 inguinal testes (5%), to the lateral or upper scrotum in 132 (31%) or around the inguinal ring in 268 (63%). In cases of high scrotal testis the gubernaculum was attached usually to the bottom of the scrotum (28, 17%), to the lateral or upper scrotum (109, 66%) or around the inguinal ring (26, 16%). CONCLUSIONS: The locations of distal gubernacular attachments in the cryptorchid testis were usually abnormal (93%). In these cases the possible testicular descent induced by hormonal therapy may either be insufficient or unable to prevent future ascent. Thus, early surgical correction rather than hormonal therapy is warranted in boys with cryptorchid testis. PMID- 15126869 TI - Volumetric and histological findings in intra-abdominal testes before and after division of spermatic vessels. AB - PURPOSE: Increased use of video laparoscopy in the diagnosis and treatment of the impalpable testis has encouraged use of the 2-stage Fowler-Stephens orchiopexy. To date, however, few limited studies exist to indicate whether clipping and division of the spermatic vessels alone may cause histological abnormalities in the intra-abdominal testis. MATERIALS AND METHODS: We evaluated histology and volume of 44 intra-abdominal testes in 35 patients between 4 months and 14 years old at stages 1 and 2 of the Fowler-Stephens procedure. RESULTS: There was a significant reduction in the number of spermatogonia and seminiferous tubules 6 months after ligation and division of the spermatic vessels. No differences were found in the number of Sertoli cells or testicular volume before and after clipping and division of the spermatic vessels. CONCLUSIONS: Ligation of the spermatic vessels during stage 1 orchiopexy for intra-abdominal testicles is associated with a significant reduction of spermatogonia. However, no significant changes were observed in the volumetric characteristics of the testicles. Further studies are necessary to evaluate the repercussions of these changes in future fertility. PMID- 15126872 TI - Urinary tract biomaterials. AB - PURPOSE: As a result of endourological advances, biomaterials have become increasingly used within the urinary tract. This review article provides an update on the current status of urinary tract biomaterials, discussing issues of biocompatibility, biomaterials available for use, clinical applications and biomaterial related complications. Perspectives on future materials for use in the urinary tract are also provided. MATERIALS AND METHODS: We performed a comprehensive search of the peer reviewed literature on all aspects of biomaterials in the urinary tract using PubMed and MEDLINE. All pertinent articles were reviewed in detail. RESULTS: Any potential biomaterial must undergo rigorous physical and biocompatibility testing prior to its commercialization and use in humans. There are currently many different bulk materials and coatings available for the manufacturing of biomaterials, although the ideal material has yet to be discovered. For use in the urinary tract, biomaterials may be formed into devices, including ureteral and urethral stents, urethral catheters and percutaneous nephrostomy tubes. Despite significant advances in basic science research involving biocompatibility issues and biofilm formation, infection and encrustation remain associated with the use of biomaterials in the urinary tract and, therefore, limit their long-term indwelling time. CONCLUSIONS: Prosthetic devices formed from biomaterials will continue to be an essential tool in the practicing urologist's armamentarium. Ongoing research is essential to optimize biocompatibility and decrease biomaterial related complications such as infection and encrustation within the urinary tract. Future advances include biodegradables, novel coatings and tissue engineering. PMID- 15126873 TI - Expression of vascular endothelial growth factor and its receptors Flk-1 and Flt 1 during the regeneration of autotransplanted adrenal cortex in the adrenalectomized rat. AB - PURPOSE: Autotransplantation of the adrenal cortex may be a therapeutic alternative in the future. For successful adrenal transplantation revascularization is necessary. It is possible that vascular endothelial growth factor (VEGF), which is a potent angiogenic peptide, may have some roles in adrenal transplantation through 2 its receptors, kinase insert domain-containing region (Flk-1) and fms-like tyrosine kinase (Flt-1). Therefore, we studied sequential changes in expression of VEGF, Flk-1 and Flt-1 in regenerated adrenal. MATERIALS AND METHODS: Eight to 9-week-old male Wistar rats underwent bilateral adrenalectomy and immediate adrenal capsular autotransplantation. The expression of VEGF, Flk-1 and Flt-1 was analyzed by immunohistochemistry and reverse transcriptase-polymerase chain reaction. RESULTS: Angiogenesis was observed in the remodeling of adrenal sinusoidal endothelium during adrenal regeneration. Reverse transcriptase-polymerase chain reaction and immunohistochemistry showed that VEGF expression increased in grafted tissue with time after transplantation and its Flk-1 receptor, which localized to endothelial cells, increased transiently during the regeneration process. Immunostaining for Flt-1 receptor was identified in adrenocortical cells and its intensity gradually increased during adrenal regeneration. CONCLUSIONS: During adrenal gland regeneration VEGF and its receptors Flk-1 and Flt-1 are thought to be involved in neovascularization. PMID- 15126874 TI - Laparoscopic mid sagittal hemicystectomy and bladder reconstruction with small intestinal submucosa and reimplantation of ureter into small intestinal submucosa: 1-year followup. AB - PURPOSE: We evaluated the long-term results of laparoscopic hemicystectomy and bladder replacement with small intestinal submucosa (SIS) with ureteral reimplantation into the SIS material. MATERIALS AND METHODS: A total of 12 minipigs underwent laparoscopic hemicystectomy. Six pigs underwent bladder reconstruction with SIS and ipsilateral ureteral reimplantation. The remaining 6 control pigs underwent hemicystectomy and primary bladder closure with ipsilateral nephroureterectomy. Preoperative and followup evaluations included blood chemistry, radiography and urodynamic evaluations. The 6, 3, 6 and 9-week, and 12-month followup evaluations included biopsies. At 1 year the animals were sacrificed. Histopathological and contractility studies, and reverse transcriptase-polymerase chain reaction for growth factors and basement membrane components were performed. RESULTS: Bladder capacity and bladder compliance were similar in the 2 groups at all time points. One pig per group died, that is a control at the 9-month evaluation due to an anesthetic complication and an SIS pig 7 months after bladder reconstruction due to spontaneous bladder rupture at the anastomotic site. In the SIS group 4 of 5 surviving pigs had unobstructed reimplanted ureters without evidence of hydroureteronephrosis, while 1 had high grade obstruction at the reimplantation site. Histopathology study after 1 year revealed muscle at the graft periphery and center but it consisted of small fused bundles with significant fibrosis. Nerves were present at the graft periphery and center but they were decreased in number. CONCLUSIONS: Laparoscopic SIS bladder reconstruction and ureteral reimplantation into the SIS after hemicystectomy are technically feasible. However, compared to primary bladder closure no advantage in bladder capacity or compliance was documented. PMID- 15126875 TI - Simultaneous expression of T-cell activating antigens in renal cell carcinoma: implications for specific immunotherapy. AB - PURPOSE: The activation of antigen specific T cells by tumor associated antigens (TAA) might be a promising treatment strategy for patients with renal cell carcinoma (RCC). We analyzed TAA expression in patients with RCC as well as the prevalence of fitting HLA phenotypes and calculated the percent of patients eligible for peptide vaccination trials. MATERIALS AND METHODS: A total of 41 RCC samples from primary tumors were analyzed for TAA expression by reverse transcriptase-polymerase chain reaction. Genes of interest were MAGE-1, MAGE-3, G250 and PRAME since peptides derived from these genes have been shown to activate antigen specific cytotoxic T lymphocytes. Results were combined with data on the HLA gene and haplotype frequencies in the German population as an example of a white population. RESULTS: Tumor specific expression of at least 1 T cell activating antigen was observed in all patients. Of the patients 80% expressed 2 or more TAAs simultaneously. HLA molecules suitable for presentation of the respective antigens were calculated to be expressed in 51% to 85% of white German patients. These results mirror with only minor variations most of the white populations in Europe and North America. CONCLUSIONS: We noted that T-cell activating tumor associated antigens are frequently expressed in patients with RCC. Based on HLA expression analysis in a white population at least 30% of patients with RCC are eligible for monovalent specific immunotherapy and 41% are eligible for polyvalent specific immunotherapy. These data are a rational basis for future prospective vaccination trials in patients with RCC. PMID- 15126876 TI - Prognostic value of carbonic anhydrase IX and KI67 as predictors of survival for renal clear cell carcinoma. AB - PURPOSE: The natural history of renal cell carcinoma (RCC) is complex and not entirely explained by conventional prognostic factors. In this study we evaluated the prognostic value of carbonic anhydrase IX (CAIX) and Ki67 to predict survival in RCC. MATERIALS AND METHODS: Immunohistochemical analysis using a CAIX and a Ki67 monoclonal antibody was performed on tissue microarrays constructed from paraffin embedded specimens from 224 patients treated with nephrectomy for clear cell renal carcinoma. CAIX and Ki67 staining were correlated with clinical factors, pathological features and survival. Median followup was 34 months (range 0.3 to 117) and disease specific survival was the primary end point assessed. RESULTS: Univariate statistical analysis showed that high Ki67 staining and low CAIX staining correlated significantly with poor median survival (21 months, p < 0.001 and 22 months, p = 0.011, respectively). Each marker was highly significant for stratifying patient groups defined by T stage, Fuhrman grade, nodal status, metastatic status and performance status. On multivariate analysis CAIX and Ki67 were significant predictors of survival with an HR of 1.78 (p = 0.014) and 1.75 (p = 0.009), respectively. Although CAIX and Ki67 staining were inversely correlated (p = 0.009), Ki67 significantly substratified patient subgroups defined by high or low CAIX staining (p = 0.001 and 0.003, respectively). When Ki67 and CAIX were combined into a single parameter, RCC tumors could be stratified into low, intermediate and high risk groups with a median survival of greater than 101, 31 and 9 months, respectively (p <0.001). On multivariate analysis the combined parameter consisting of Ki67 and CAIX was a significant predictor of survival (p <0.001) and it was able to displace histological grade. CONCLUSIONS: Ki67and CAIX are useful prognostic biomarkers for RCC that improve the survival prediction and classification of kidney cancer. PMID- 15126877 TI - Suppression of lung metastasis of renal cell carcinoma by the intramuscular gene transfer of a soluble form of vascular endothelial growth factor receptor I. AB - PURPOSE: We clarified whether adenovirus mediated intramuscular gene transfer of soluble vascular endothelial growth factor receptor 1 (sFlt-1) can inhibit lung metastasis of renal cell carcinoma. MATERIALS AND METHODS: We constructed the adenovirus vector AdsFlt-1, which expresses soluble Flt-1 protein. Functional validation of the vector was determined by HUVEC (in vitro human umbilical vein endothelial cell) proliferation inhibition assay. The efficacy of AdsFlt-1 was tested in a Renca murine renal cell carcinoma lung metastasis model. BALB/c mice were injected with Renca cells, followed by the intramuscular administration of AdsFlt-1 24 hours later. Lung specimens were harvested 17 days later and the number of lung metastases was counted. Immunohistochemical analysis of the specimen for angiogenesis and apoptosis was done. RESULTS: Treatment with adenovirus expressed sFlt-1 inhibited HUVEC proliferation. The intramuscular administration of AdsFlt-1 significantly inhibited lung metastasis of Renca cells. Immunohistochemical analysis of the lung specimen showed fewer neovessel formations and more apoptotic cells in AdsFlt-1 treated tumors in mice. CONCLUSIONS: The intramuscular administration of AdsFlt-1 effectively inhibited lung metastasis in a murine model of renal cell carcinoma. Because of the simplicity of this therapy, it may well be useful as an effective adjuvant therapy for renal cell carcinoma. PMID- 15126878 TI - Systematic in vitro evaluation of survivin directed antisense oligodeoxynucleotides in bladder cancer cells. AB - PURPOSE: The rather poor responses to conventional treatment for bladder cancer (BCa) require novel, specific therapy approaches. The down-regulation of BCa associated genes may represent a new option to inhibit specifically BCa cell growth and induce cell death. Survivin, an apoptosis inhibitor that is up regulated in the majority of malignancies, including BCa, provides an attractive target for molecular therapies, such as treatment with specific antisense oligode oxynucleotides (AS-ODNs). MATERIALS AND METHODS: We used mRNA secondary structure prediction to design survivin directed AS-ODNs. After lipid mediated transfection with 30 selected antisurvivin AS-ODN inhibitory effects on cell growth properties as well as on survivin expression were measured. RESULTS: Three of 30 tested constructs reproducibly impaired the growth characteristics of 4 BCa cell lines. Detailed analysis of the cell line EJ28 treated with the constructs SVV261, SVV264 and SVV286 revealed a clear decrease in viability (down to 35%) and long term proliferation (down to 14%), which were caused by cell cycle arrest and an increase in apoptosis (from 19.5% to 51.3% maximum). The inhibition of tumor cell growth was associated with up to 60% to 80% survivin expression down-regulation. Interestingly all 3 evolved AS-ODNs were directed against the putative single strand survivin mRNA motif between 274 to 285 nucleotides, identified by secondary structure prediction. The reported accessibility of this motif to other nucleic acid based inhibitors such as ribozymes and small interfering RNAs emphasizes the rationale of a systematic selection of mRNA target sites. CONCLUSIONS: The survivin directed AS-ODNs shown to inhibit effectively the proliferation of BCa cells in the current study may provide suitable adjuvant therapeutic agents for the specific local treatment of BCa. PMID- 15126879 TI - Synergistic antitumor activity by combined treatment with gemcitabine and antisense oligodeoxynucleotide targeting clusterin gene in an intravesical administration model against human bladder cancer kotcc-1 cells. AB - PURPOSE: We investigated whether antisense (AS) oligodeoxynucleotide (ODN) targeting the clusterin gene enhances the cytotoxic effect of gemcitabine in human bladder cancer KoTCC-1 cells in vitro and in vivo, and evaluated the usefulness of the combined administration of AS clusterin ODN and gemcitabine using an intraperitoneal tumor cell injection model. MATERIALS AND METHODS: The cytotoxic effect of combined treatment with AS clusterin ODN and gemcitabine on in vitro KoTCC-1 growth was examined using the MTT (3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide) assay. The in vivo growth inhibitory effects of combined AS clusterin ODN and gemcitabine therapy on subcutaneous KoTCC-1 tumor was also examined. The intraperitoneal tumor cell injection model, which mimics intravesical administration therapy against bladder cancer, was used to evaluate the efficacy of combined AS clusterin ODN and gemcitabine therapy. RESULTS: AS clusterin ODN treatment of KoTCC-1 cells significantly enhanced gemcitabine chemosensitivity in a dose dependent manner, decreasing gemcitabine IC50 by approximately 90%. In vivo systemic administration of AS clusterin ODN and gemcitabine significantly decreased subcutaneous KoTCC-1 tumor volume compared with scramble control ODN plus gemcitabine. Furthermore, combined administration of AS clusterin ODN plus gemcitabine resulted in significantly delayed formation of hemorrhagic ascites compared with scramble control ODN plus gemcitabine in an intraperitoneal tumor cell injection model. CONCLUSIONS: These findings suggest that AS clusterin ODN may be useful for enhancing the cytotoxicity of gemcitabine in patients with bladder cancer, particularly as a novel therapeutic strategy for intravesical instillation therapy. PMID- 15126880 TI - Management of bladder cancer. PMID- 15126881 TI - Asp85tyr polymorphism in the udp-glucuronosyltransferase (UGT) 2B15 gene and the risk of prostate cancer. AB - PURPOSE: An amino acid changing polymorphism in the UDP-glucuronosyltransferase (UGT) 2B15 gene has been described at codon 85 (aspartate>tyrosine). The UGT2B15 allele exhibits 2-fold decreased activity for dihydrotestosterone, which was suggested to be associated with the risk of prostate cancer based on in vitro functional analysis. We determined whether this polymorphism can be used to predict individual susceptibility to prostate cancer. MATERIALS AND METHODS: UGT2B15 expression was determined by reverse transcription-polymerase chain reaction of normal prostate tissues. Prevalence of the UGT2B15 Asp85Tyr polymorphism was compared between cases and controls by allele specific polymerase chain reaction analysis using genomic DNA isolated from 155 incident white patients with primary prostate cancer and 155 individually age matched (+/ 5 years) white controls. RESULTS: UGT2B15 mRNA was detected in all prostate tissues tested. A significant association was found between UGT2B15 genotypes and prostate cancer risk. A significantly increased risk of prostate cancer was observed in subjects with the homozygous UGT2B15 genotype (OR 2.7, 95% CI 1.1 to 6.6). CONCLUSIONS: These results suggest that the UGT2B15 enzyme may have a role in the metabolism of dihydrotestosterone in prostate tissue and UGT2B15 Asp85Tyr polymorphism is associated with prostate cancer risk. PMID- 15126882 TI - 4 layer versus 1 layer small intestinal submucosa for correction of penile chordee: experimental study in a rabbit model. AB - PURPOSE: We evaluated the use of 4 vs 1 layer small intestinal submucosa (SIS) for covering defects in the ventral surface of the tunica albuginea to correct severe chordee. MATERIALS AND METHODS: A total of 18 New Zealand white rabbits underwent implantation of a 10 x 5 mm SIS graft following excision of a rectangular area in the ventral surface of the tunica albuginea. In 9 rabbits 4 layer SIS was used to cover the defect and in the remaining animals 1 layer SIS was used. The animals were sacrificed at 2, 6 and 12-week intervals postoperatively, respectively. The surface area of the grafts was measured and the percent of contracture was calculated. Transverse sections of the penis at the graft site were stained with hematoxylin and eosin, and Masson's trichrome, and examined microscopically. RESULTS: : None of the animals had hematoma or bleeding. At autopsy contracture was not seen in any of the rabbits with 1 layer SIS. On the contrary, there was 21% and 25% contracture at 6 and 12 weeks, respectively, in the 4 layer SIS group. At 12 weeks the 1 layer SIS graft was completely replaced by well collagenized tissue similar to that of normal tunica albuginea without inflammatory infiltrate, while the multilayer SIS graft was replaced by dense fibrous tissue with areas of chronic inflammation and other focal areas of calcification. CONCLUSIONS: Four layer SIS undergoes contracture and calcification when used to cover defects in the tunica albuginea. On the other hand, 1 layer SIS can be safely and reliably used for corporeal grafting. PMID- 15126883 TI - Identification of kit positive cells in the human urinary tract. AB - PURPOSE: Analogous to interstitial cells of Cajal in the bowel, functional important networks of interstitial cells could have a role in the complex mechanism of central and peripheral control of urinary tract function. Recently various reports mentioned the presence of interstitial cells in different parts of the urinary tract and in different species. Since important differences among species exist, we performed immunohistochemistry on fresh frozen human tissue to study the presence of interstitial cells in the human urinary tract. MATERIALS AND METHODS: A total of 65 tissue pieces from all levels of the urinary tract were obtained from 44 patients treated at our institution. Tissue was processed for immunohistochemistry immediately after removal. We performed immunohistochemistry for kit, connexin 43 and VRL1/TRPV2. RESULTS: Interstitial cells immunopositive for all 3 antibodies were seen beneath the urothelium and between smooth muscle cells in all tissue pieces with slight topographical differences. CONCLUSIONS: Together with morphological and functional data from other experiments these morphological data suggest that, as in the bowel, networks of interstitial cells might have an important role in the physiology and pathology of the urinary tract. They could be involved in pacemaking or have an integrating role through the modulation of neurotransmission and conduction of electrical impulses. Functional experiments are the next step to study these hypotheses. PMID- 15126884 TI - Psychosexual outcome in women affected by congenital adrenal hyperplasia due to 21-hydroxylase deficiency. AB - PURPOSE: Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency results in increased adrenal androgen secretion. When the deficiency is severe, the result is the salt losing (SL) form, and when the deficiency is partial, the result is the simple virilizing (SV) form of CAH. We documented long-term satisfaction with gender, cosmetic appearance and function of the genitalia, and surgical management practices in a group of women with CAH due to 21-hydroxylase deficiency. MATERIALS AND METHODS: Psychosexual and surgical outcome were assessed in 41 women with CAH using an interview, a written questionnaire and a physical examination. When appropriate, outcome measures were compared to those in unaffected control women. RESULTS: Women in the SL group were more likely to question their female gender and report sexual concerns, and less likely to have sexual relations with a partner than those with the SV form or control women. Overall women with CAH were moderately satisfied with the cosmetic appearance of the genitalia but the SL group reported worse genital function than the SV group. Physician rated appearance of the genitalia was better than ratings provided by patients and women with the SL form were judged to have a worse cosmetic outcome of genital reconstruction than women with the SV form. The most common response concerning the optimal timing for genital reconstruction was during infancy and early childhood, although a number of women favored waiting until later for these procedures or did not respond to this question. CONCLUSIONS: Women with the SV form reported greater satisfaction and fewer concerns regarding their psychosexual and surgical outcome than women with the SL form. PMID- 15126885 TI - Experimental comparison of high power (80 W) potassium titanyl phosphate laser vaporization and transurethral resection of the prostate. AB - PURPOSE: Perioperative hemorrhage is still the major complication of standard transurethral prostate resection (TURP). Potassium titanyl phosphate (KTP) laser vaporization using 80 W is a novel technique that promises instant hemostatic tissue ablation. In this ex vivo investigation we compared the hemostatic properties of the 2 procedures. MATERIALS AND METHODS: Ex vivo, blood perfused porcine kidneys were used to verify the hemostatic efficacy of KTP laser vaporization and TURP-like tissue resection. Bleeding could be exactly quantified in relation to tissue ablation for the 2 techniques. In addition, specimens were examined microscopically. RESULTS: KTP laser vaporization demonstrated highly significantly decreased bleeding as compared to conventional tissue resection for a standardized ablation volume of 16 cm tissue (2.1 vs 23.3 ml per minute, p <0.0001). Tissue ablation was more rapid in the resection group (20 vs 100 seconds, p <0.001). Histological examinations revealed larger coagulation zones for the KTP group compared to conventional tissue resection (0.9 vs 0.6 mm, p <0.01). CONCLUSIONS: Ex vivo, 80 W KTP laser vaporization is a virtually bloodless ablative procedure, giving rise to hemostasis that is highly superior to conventional TURP-like tissue resection. However, the novel procedure is considerably more time-consuming. PMID- 15126887 TI - Somatoform dissociation and adverse childhood experiences in the general population. AB - Childhood trauma has been associated with psychological dissociation, but there is evidence that trauma may also result in somatoform dissociation. We performed a general population study with 1739 subjects, using the Somatoform Dissociation Questionnaire, measures of adverse childhood experiences, and sociodemographic background. The prevalence of high somatoform dissociation (Somatoform Dissociation Questionnaire >or=30) was 9.4% in the Finnish general population. Unemployment, a reduced working ability, and a poor financial situation were associated with high somatoform dissociation. Of the adverse childhood experiences, high somatoform dissociation was strongly linked to physical punishment but not associated with domestic violence, including sexual and physical abuse. The odds of high somatoform dissociation were also increased among men by a poor relationship between their parents, and among women by alcohol abuse in their childhood home. We found a strong, graded relationship between an increasing number of adverse childhood experiences and high somatoform dissociation. PMID- 15126888 TI - Instruments for the assessment of childhood trauma in adults. AB - The relationship between childhood trauma and adult psychopathology has been explored in the literature. The goal of this study is to compare existing instruments that measure retrospective interpersonal gross childhood trauma. A computerized search from 1985 to March 2003 was performed to locate instruments used to measure childhood trauma. These were divided into interview-rated and self-report measures and were compared on various parameters. Twenty-one observer rated and 21 self-report instruments were identified. In a comparison, five observer-rated measures and three self-report measures stand out for having favorable characteristics such as assessing multiple types of trauma and reporting on psychometric properties. A number of instruments are designed to measure a single type of trauma, usually sexual abuse, but most of these do not report psychometric properties. A few instruments used to measure retrospective childhood trauma are particularly useful for systematic research in adult psychiatric disorders. PMID- 15126889 TI - Agreement between self-rated and clinically assessed symptoms in subjects with psychosis. AB - The aim of this study was to explore the capacity of acutely ill patients with psychosis (N = 40) to self-report their symptoms by comparing self-assessment and objective measures. Positive, negative, and depressive symptoms were rated using the Scale for the Assessment of Positive Symptoms, the Scale for the Assessment of Negative Symptoms, and the Calgary Depression Scale. Insight level was measured using the Scale to Assess Unawareness of Mental Disorder. Patients were asked to self-report positive, negative, and depressive symptoms using the Community Assessment of Psychic Experience. Patients presenting with acute psychotic disorders are able to assess fairly their positive, negative, and depressive symptoms. Significant associations were found between self-reported and objective measures of positive, negative, and depressive symptoms independently of insight level. Individual positive and negative symptoms were correctly self-assessed, except for persecutory delusion and alogia, respectively. These results suggest that self-report questionnaires can be used in educational programs to favor the patient's therapeutic adherence. PMID- 15126890 TI - Factors associated with compliance and resistance to command hallucinations. AB - Command hallucinations (CHs) are hallucinations that direct the patient to perform an action. Beyond issues related to the danger that some CHs may pose, comparatively little is known about the broader clinical context of CHs. To investigate this, 199 patients were interviewed using the Mental Health Research Institute Unusual Perceptions Scale. More than two thirds of the sample reported hearing CHs. A quarter of these patients felt unable to resist them. Patients with CHs reported their voices more negatively than those who did not hear commands. More of those unable to resist CHs rated their hallucinations as intrusive, they had fewer coping strategies than those able to resist, and they were prescribed higher dosages of medication. CHs are associated with a greater degree of adverse hallucinatory and illness experience. Patients who experience CHs may have a more malignant form of the underlying disorder. Thus, CHs warrant special therapeutic attention for reasons beyond any harm that their commands pose to themselves or others. PMID- 15126891 TI - Determinants of depression among Ethiopian immigrants and refugees in Toronto. AB - The purpose of this study was to determine the occurrence of and risk factors for depressive disorder in a random sample of 342 Ethiopian immigrants and refugees in Toronto. The Composite International Diagnostic Interview questionnaire was used to measure depression. The results suggested a lifetime prevalence of depression among Ethiopian immigrants and refugees of 9.8%, which was slightly higher than the lifetime prevalence rate in the Ontario population (7.3%). However, the rate among Ethiopian immigrants and refugees was approximately three times higher than the rate estimated for Southern Ethiopia (3.2%). The data confirmed the significance of known risk factors for depression in immigrants, including younger age, experiences of premigration trauma, refugee camp internment, and postmigration stressful events. The implication of the overall finding is that there is a need to develop mental health intervention programs, particularly for people who have experienced premigration trauma, refugee camp internment, and postmigration stresses. PMID- 15126892 TI - Life events, social support, and onset of major depressive episode in Finnish patients. AB - We investigated differences in life events and social support between subgroups of depressed patients and the distribution of life events in phases preceding or during depression. In the Vantaa Depression Study, 269 psychiatric patients with DSM-IV major depressive disorder were diagnosed with Schedule for Clinical Assessment in Neuropsychiatry, Version 2.0, and Structured Clinical Interview for DSM-III-R personality disorders (SCID-II). Life events during the 12 months preceding the interview were investigated with the Interview for Recent Life Events, and social support with the Interview Measure of Social Relationships and the Perceived Social Support Scale-Revised. Nearly all patients (91%) reported life events, on average 4.1 per preceding year. No major differences between sociodemographic or clinical subgroups were found; the frequency of events was somewhat greater among the younger subjects, whereas those with comorbid alcoholism or personality disorders perceived less social support. Although events were distributed evenly between the time preceding depression, the prodromal phase, and the index major depressive episode, two thirds of the patients attributed their depression to some event. Despite clinical and sociodemographic heterogeneity, patients with major depressive disorder are fairly homogeneous in terms of life events during the preceding year. Events do not cluster in any particular phase of the progression to an episode. PMID- 15126893 TI - Neuropsychological impairment associated with symptoms of schizotypy: role of depressive and paranoid symptoms. AB - : Little is known about the role of depressive symptoms in cognitive impairment associated with schizotypy. We compared neuropsychological performance among people reporting high levels of both schizotypal symptomatology on the Schizotypal Personality Questionnaire (SPQ) and depressive symptoms on the Beck Depression Inventory-II (BDI-II), people with high SPQ and low BDI-II, and healthy controls (low SPQ and BDI-II). We hypothesized that the group with both depressive and schizotypal features would demonstrate the greatest impairment. However, that group demonstrated better verbal memory and cognitive set maintenance. Further examination revealed higher endorsement of paranoid/suspicious symptoms in the people with both depressive and schizotypal features. Results highlight the importance of distinguishing among subtype variations in schizotypal research. PMID- 15126894 TI - Cohabitation, education, and occupation of psychiatric outpatients bullied as children. AB - Few studies have explored adult psychiatric outpatients who were bullied in childhood. The aim of this study was to contrast social-demographic variables of adult psychiatric outpatients who reported bullying in childhood with those without such reports. One hundred sixty consecutive adult patients from a psychiatric outpatient clinic completed self-administered questionnaires about cohabitation status, level of education, work status, and occupation. Bullying was measured by an inventory used in schools. The results showed that psychiatric outpatients bullied in childhood were more often singles, had significantly lower levels of education, often received social benefits, and worked as shop assistants rather than as engineers and schoolteachers. Those bullied in childhood showed poorer psychosocial adjustment as adults. PMID- 15126895 TI - Relationship of alcohol craving to symptoms of posttraumatic stress disorder in combat veterans. AB - To examine the relationship of alcohol craving to symptoms of posttraumatic stress disorder (PTSD), 129 male veterans with chronic PTSD were asked to complete the Obsessive Compulsive Drinking Scale (OCDS), the Mississippi Scale for combat-related PTSD symptoms, and other instruments to assess general psychopathology and lifetime alcohol and substance use. No correlations were found between current PTSD symptoms and alcohol craving, although significant correlations were found between the OCDS and measures of lifetime alcohol and substance use. PMID- 15126900 TI - Genetics of primary aldosteronism. AB - Over the last few years, much progress has been made in understanding the genetic basis of primary aldosteronism. This has led to the diagnosis of the familial forms and has aided the understanding of the basis of sporadic forms of the disease. Such information can be exploited to improve the therapeutic approaches used not only for patients with primary aldosteronism, but also with other forms of hypertension. Here, we review the genetic and the phenotypic features of the familial forms, the genetic variants that influence the sporadic forms and the structure and regulation of the CYP11B1 and CYP11B2 genes. PMID- 15126901 TI - Low blood pressure and low energy: (how) are they related? PMID- 15126902 TI - Endothelin-1 and blood pressure in women. PMID- 15126903 TI - Nitric oxide and vascular hypertrophy. PMID- 15126904 TI - G protein alpha(i2): a potential link between hypertension and insulin resistance? PMID- 15126905 TI - Aspirin, superoxide anions and development of hypertension. PMID- 15126906 TI - Linkage of Na(+) and Ca(2+) balance: evidence that Na(+) retention preserves Ca(2+) balance and limits bone wasting. PMID- 15126907 TI - Angiotensin, high blood pressure and glomerular autoregulation. PMID- 15126908 TI - The SYST-EUR study and beyond: immediate versus delayed treatment of isolated systolic hypertension. PMID- 15126909 TI - Prospective association between hypotension and idiopathic chronic fatigue. AB - OBJECTIVE: To determine whether there is an association between hypotension and incident cases of idiopathic chronic fatigue. DESIGN: A prospective study. SETTING: Johns Hopkins Precursors Study. PARTICIPANTS: Medical students (n = 876) in graduating classes from 1948 to 1964. MAIN OUTCOME MEASURE: 'Easy fatigability' reported by participants at 5- or 10-year follow-up after graduation. RESULTS: The unadjusted risk in women was 5.0 (95% exact confidence interval = 1.4 to 17.4) and in men was 1.7 (95% exact confidence interval = 0.8, to 3.5). CONCLUSION: These preliminary findings suggest that hypotension may be a risk factor for the development of idiopathic chronic fatigue in women. PMID- 15126910 TI - Dietary sodium and pulse pressure in normotensive and essential hypertensive subjects. AB - OBJECTIVES: To assess the influence of sodium intake on the two components of arterial blood pressure [mean blood pressure (MBP) and pulse pressure] and left ventricular mass (LVM) in normotensive subjects and never-treated hypertensive patients. METHODS: Blood pressure, LVM (M-mode echocardiography) and sodium excretion as an index of dietary sodium were measured in 459 males and 396 females, aged 14-93 years, with elevated (63% of population) or normal blood pressure. RESULTS: Because pulse pressure decreased with age up to approximately 40 years of age, and increased thereafter, analysis was undertaken separately in 438 young (age < or = 40 years) and 417 middle-aged to elderly subjects. Before age 40 years, only pulse pressure was independently and positively influenced by urinary sodium excretion. After age 40 years, MBP and LVM index but not pulse pressure were positively influenced by urinary sodium excretion. When men and women were analysed separately according to tertiles of urinary sodium excretion, pulse pressure adjusted for MBP and age increased from the first to the third tertile of urinary sodium excretion in males (P < 0.03), but not in females. CONCLUSIONS: The influence of sodium intake on pulse pressure was found in the younger subjects of both males and females and only in men older than 40 years. PMID- 15126911 TI - Cellular adhesion molecules and blood pressure: interaction with sex in a multi ethnic population. AB - OBJECTIVE: To clarify the association between blood pressure and four different adhesion molecules, adjusting for potential confounders, in men and women from different ethnic origins. DESIGN AND METHODS: The soluble (s) plasma adhesion molecules sP-selectin, sE-selectin, intracellular adhesion molecule-1 (sICAM-1) and vascular cell adhesion molecule-1 (sVCAM-1) were measured in 261 white (120 women), 188 African origin (99 women) and 215 South Asian (99 women) individuals living in England. All were free from coronary heart disease, stroke, other cardiovascular disease and diabetes, and were not receiving drug treatment for hypertension or high lipids, hormone replacement therapy or oral contraceptives. RESULTS: After adjustment for age, only sE-selectin concentrations were significantly associated with blood pressure. There was a significant interaction of sex with systolic (P = 0.013), diastolic (P = 0.042) and pulse (P = 0.015) pressures. After adjustment for age, ethnicity, body mass index and smoking, the significant interaction of sex persisted and in women the associations with systolic (P < 0.001), diastolic (P < 0.001) or pulse (P = 0.004) pressure were unchanged, but in men the association with diastolic blood pressure was abolished. Finally, the association appeared to be present in women younger than 50 years, who were likely to be premenopausal. CONCLUSIONS: The relationship between adhesion molecules and blood pressure is adhesion molecule specific and varies with sex and age, which may partially explain previous inconsistencies in the literature. The mechanisms relating blood pressure to adhesion molecule concentrations are unknown, but they are likely to be modified by the menopause. These differences may relate to the production, clearance or cell-surface shedding of the adhesion molecules. PMID- 15126912 TI - Plasma endothelin-1 level is related to renal function and smoking status but not to blood pressure: an epidemiological study. AB - OBJECTIVE: Endothelin-1 (ET-1) is a potent vasoconstrictor derived from the endothelium. Several studies with small numbers of humans have showed high plasma ET-1 levels in hypertension, but other studies have not. Furthermore, it has been shown in a small number of subjects that ET-1 is elevated in uraemic patients. However, there have been no epidemiological surveys as to whether ET-1 level is related to hypertension or end-organ damage. METHODS: A total of 1492 subjects received a health examination in 1999. The data for fasting ET-1 of 1450 individuals were obtained. A specific radioimmunoassay was used to measure ET-1 levels. We also measured body mass index (BMI), systolic and diastolic blood pressure (BP), haemoglobin A1c, cholesterol, blood urea nitrogen (BUN), creatinine and uric acid. We performed carotid B-mode ultrasonography and electrocardiography. Smoking habit was evaluated by questionnaire. RESULTS: Mean ET-1 was 4.93 +/- 1.73 pg/ml in men and 4.84 +/-1.54 pg/ml in women. ET-1 increased with age (P < 0.001). Systolic (P < 0.001) and diastolic (P < 0.05) BP, hypertensive medication (P < 0.05), BUN (P < 0.01), creatinine (P < 0.001), uric acid (P < 0.001), intimal-medial thickness (P < 0.001), smoking (P < 0.05) and age (P < 0.001), were significantly associated with ET-1 by univariate analysis. By the use of multiple stepwise regression analysis, age (P < 0.001), creatinine (P < 0.001) and smoking (P < 0.05) remained significant. However, no relation was shown between ET-1 and BP. CONCLUSION: Our data suggest that high ET-1 is not related to hypertension, but to subclinical renal dysfunction and smoking. PMID- 15126913 TI - Age-dependent associations between blood pressure and coronary artery calcification in asymptomatic adults. AB - OBJECTIVE: The age-dependent roles of the components of blood pressure (BP) in the development of coronary artery calcification (CAC) are poorly understood. DESIGN: We examined systolic (SBP), diastolic (DBP), mean arterial pressure (MAP) and pulse pressure (PP) as predictors of CAC in 830 asymptomatic, non-diabetic participants in a community-based study who were aged > or = 30 years and free of antihypertensive therapy or known cardiovascular disease. METHODS: CAC was measured with electron beam computed tomography. Tobit regression was used in two age groups (< 50 years and > or = 50 years) to evaluate the relationship of BP components with presence and quantity of CAC, adjusting for traditional coronary artery disease (CAD) risk factors. RESULTS: Among those aged < 50 years, CAC was positively associated with SBP, DBP and MAP, considering each pressure individually and DBP was the strongest predictor (P = 0.0088). Among those aged > or = 50 years, CAC was positively associated with SBP (P = 0.0257) and PP (P = 0.0028), considered individually. When SBP and DBP were in the same model, presence and CAC quantity were positively associated with SBP (P = 0.0024) and negatively with DBP (P = 0.0401), favoring PP as the best predictor of CAC. CONCLUSIONS: SBP, DBP and PP have age-dependent roles in the prediction of CAC similar to their roles in prediction of future CAD events. These observations provide new evidence supporting the measurement of CAC as a surrogate of target organ disease and subsequently, as a predictor of increased risk of future CAD events. PMID- 15126914 TI - Contrasting circadian rhythms of blood pressure among inbred rat strains: recognition of dipper and non-dipper patterns. AB - OBJECTIVE: The non-dipper pattern, i.e. the lack of nocturnal blood pressure (BP) fall, carries a high risk of cardiovascular complications, both in hypertensive and normotensive subjects. Without genetic engineering, experimental demonstration of the non-dipper phenomenon is lacking. The purpose of this study was to assess the haemodynamic and behavioural daily parameters among various strains of rats - spontaneously hypertensive rats (SHR), Wistar-Kyoto (WKY) and Fischer 344 (F344) - in order to characterize their circadian patterns and to detect a non-dipper animal model. METHODS: Changes in BP, heart rate (HR), and spontaneous locomotor activity (SLA) were recorded continuously for 11 days using telemetry in freely moving 10-week-old male SHR, WKY and F344 rats, in standardized laboratory conditions. Variations in haemodynamic and behavioural parameters were assessed in terms of day/night differences and spectral power corresponding with the 24-h period. RESULTS: All rats exhibited clear circadian variations in HR and in SLA, in synchrony with the light cycle. Light/dark differences in BP were significantly lower in F344 compared with those of SHR and WKY. The smaller circadian changes in BP observed in F344 were also demonstrated using spectral analysis: the peak detected at 24-h was reduced in F344 compared with SHR and WKY. CONCLUSION: The inbred F344 strain lacks the typical circadian BP rhythm while oscillations of HR and SLA are maintained, suggesting different regulatory mechanisms. The F344 strain may represent a useful animal model for studying the effects of drugs aimed at restoring the dipper status. PMID- 15126915 TI - Genes encoding endothelin-converting enzyme-1 and endothelin-1 interact to influence blood pressure in women: the EVA study. AB - BACKGROUND: Endothelin-1 (ET-1) is a potent vasoactive peptide that has been implicated in the regulation of basal vascular tone. Endothelin-converting enzyme 1 (ECE-1), the main enzyme responsible for ET-1 generation, may contribute to blood pressure (BP) control. A possible association between a polymorphism of the gene encoding ECE-1 (ECE1B C-338A) and BP values in untreated hypertensive women was recently reported. OBJECTIVE: We studied the influence of the ECE1B C-338A polymorphism on BP levels in 1189 subjects participating in the Etude du Vieillissement Arteriel (EVA study), and looked for an interaction between this variant and a polymorphism of the ET-1 gene (EDN1 K198N). METHODS: The ECE1B C 338A polymorphism was genotyped in 491 men and 698 women; 477 men and 669 women could also be genotyped for the EDN1 K198N polymorphism. Associations between BP levels and genotypes were assessed by ANOVA; ANCOVA was used to control for covariates. RESULTS: We found an association between the ECE1B C-338A polymorphism and BP levels in women but not in men. Specifically, females homozygous for the A allele had significantly higher systolic, diastolic and mean BP levels (P = 0.01, 0.02, 0.006 respectively, after adjustment for age and body mass index). Genotyping of the EDN1 K198N polymorphism showed that this variant was not associated with BP values in either men or women, but interacted with the ECE1 variant to influence systolic and mean BP levels in women. CONCLUSION: Results from this large association study suggest that the genes encoding ECE-1 and ET-1 interact to modulate BP levels in women. PMID- 15126916 TI - Role of tissue kallikrein in response to flow in mouse resistance arteries. AB - BACKGROUND: Tissue kallikrein, an essential enzyme in the formation of vascular kinins, contributes to flow-dependent dilatation (FDD) in large arteries. We hypothesized that the vascular kinin-kallikrein system may be involved in shear stress signalling in small resistance arteries, which have a key role in the systemic regulation of blood pressure. OBJECTIVE: To investigate the role of the vascular kallikrein-kinin system in mesenteric resistance arteries of mice during acute changes in blood flow. DESIGN: Arteries from wild-type mice (TK) and mice lacking tissue kallikrein (TK) were mounted in an arteriograph for the recording of changes in outer diameter during step increases in flow rate. RESULTS: Responses to phenylephrine, acetylcholine or sodium nitroprusside were not different between the two strains. FDD was significantly reduced in arteries of TK mice compared with that in mesenteric arteries of TK mice exposed to phenylephrine (P = 0.04). FDD was no longer different between TK and TK mice when experiments were performed in the presence of the nitric oxide synthase (NOS) inhibitor N-nitro-l-arginine methyl ester (l-NAME; P = 0.26), l-NAME plus diclofenac (P = 0.73), or l-NAME plus diclofenac plus potassium chloride (P = 0.31), indicating that inactivation of tissue kallikrein preferentially affects the contribution of nitric oxide to flow response. However, expression of endothelial NOS was comparable between TK and TK mesenteric arteries. Finally, the bradykinin B2 receptor antagonist, HOE-140, significantly decreased FDD in TK but not in TK arteries (P = 0.03 and P = 0.82, respectively). CONCLUSION: These results demonstrate the specific role of the tissue kallikrein in flow-induced dilatation, which is mediated by nitric oxide and bradykinin B2 receptor activation in resistance arteries. PMID- 15126917 TI - Chronic l-arginine treatment reduces vascular smooth muscle cell hypertrophy through cell cycle modifications in spontaneously hypertensive rats. AB - OBJECTIVE: To investigate the effect of long-term l-arginine supplementation on phenotype and proliferative status of vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) as well as the possible changes in nitric oxide (NO) availability. METHODS: Male SHR, 22 weeks of age, received l-arginine (660 mg/kg per day) in their drinking water for 12 weeks. VSMCs from untreated (C VSMC) and l-arginine-treated (l-Arg-VSMC) SHR were isolated from the common carotid artery, cultured and used until passage five. Size, protein content, cell proliferation and ploidy were evaluated in carotid VSMCs in culture, as well as the possible association of NO in these changes. RESULTS: Relative cell size, total protein content per cell, and number of polyploid cells were significantly lower in l-Arg-VSMC compared to C-VSMC. Fetal calf serum stimulation (10% FCS) increased cell number only in l-Arg-VSMC. DNA synthesis, assessed by [H]methylthymidine incorporation after 10% FCS stimulation, was higher in l-Arg VSMC than in C-VSMC. Cell cycle analysis revealed a significant increase of the number of l-Arg-VSMC at the G1 phase, together with a reduction at the G2 + M phase. In contrast, C-VSMC were arrested at the G2 + M phase of the cell cycle. Nitrite/nitrate levels, as well as intracellular cyclic guanosine monophosphate (cGMP) content, were significantly higher in l-Arg-VSMC. This was accompanied by enhanced inducible nitric oxide synthase (iNOS) expression and activity and a decreased constitutive nitric oxide synthase (cNOS) activity in these cells. CONCLUSIONS: The results suggest that chronic treatment with l-arginine induces changes in VSMC size, ploidy and cell cycle. These changes are accompanied by iNOS induction and stimulation of the NO-cGMP pathway. PMID- 15126918 TI - Bradykinin and matrix metalloproteinases are involved the structural alterations of rat small resistance arteries with inhibition of ACE and NEP. AB - BACKGROUND AND AIM: Increased vascular resistance is a hallmark of hypertension and involves structural alterations, which may entail smooth muscle cell hypertrophy or hyperplasia, or qualitative or quantitative changes in extracellular matrix (ECM) proteins. Since the renin-angiotensin-aldosterone system modulates these changes, we investigated the effects of 8 weeks of treatment with an angiotensin-converting enzyme (ACE) inhibitor, ramipril (RAM), or a dual ACE and neutral endopeptidase (NEP) inhibitor, MDL-100240 (MDL), on mesenteric small artery structure and ECM proteins in mRen2-transgenic rats (TGRs), an animal model of hypertension with severe cardiovascular damage. MATERIALS AND METHODS: Thirty-five 5-week-old rats were included in the study: six TGRs received RAM; five TGRs RAM + the bradykinin receptor inhibitor, icatibant; six TGRs, MDL; and five TGRs MDL + icatibant, while eight TGRs and five normotensive Sprague-Dawley controls were kept untreated. Mesenteric small arteries were dissected and mounted on a micromyograph. The media-to-lumen ratio (M/L) was then calculated. Vascular metalloproteinase (MMP) content was evaluated by zymography. RESULTS: In untreated TGRs severe hypertension was associated with inward eutrophic remodelling of small arteries. Both RAM and MDL prevented the increase in blood pressure and M/L and decreased MMPs. Icatibant blunted the effect of MDL on BP, M/L and MMPs. CONCLUSIONS: Changes in collagenase activity induced by ramipril and MDL are associated with prevention of small artery structural alterations in TGRs. Furthermore, MDL-induced enhancement of bradykinin could play a role in both the prevention of vascular structural alterations and in the stimulation of MMPs. PMID- 15126919 TI - Cardiovascular hypertrophy in one-kidney, one-clip renal hypertension is resistant to heparin. AB - OBJECTIVE: Heparin inhibits vascular hypertrophy in angiotensin-induced hypertension, in addition to its well-known role in inhibiting injury-induced vascular smooth muscle proliferation. We tested whether hypertension and vascular hypertrophy could be reduced by heparin independently from the renin-angiotensin system. METHODS: Rats were made hypertensive with a one-kidney, one-clip (1K1C) procedure and received heparin from osmotic minipumps (0.3 mg/h per kg i.v.) or saline vehicle for 2 weeks. Blood pressure was measured by the tail-cuff method and vessel cross-sectional area was measured by morphometry in the aorta and mesenteric arteries. Proliferation was assessed with bromodeoxyuridine labelling. RESULTS: Blood pressure elevation and cardiovascular hypertrophy were evident in 1K1C rats. The media of mesenteric arteries was increased by 25%, and the media : lumen ratio by 35%, in hypertensive rats. DNA synthesis by smooth muscle cells in the mesenteric arteries was increased sevenfold in renal hypertension. Heparin treatment did not influence either the increase in blood pressure, the cardiovascular hypertrophy response or hypertension-mediated proliferation of arterial smooth muscle cells. CONCLUSIONS: These data suggest that the vascular hypertrophy mechanisms operating in 1K1C renal hypertension are not inhibited by heparin and thus are different from those in angiotensin-mediated hypertension. Identifying such mechanisms in the future will be important for devising appropriate intervention strategies in angiotensin-independent forms of vascular hypertrophy. PMID- 15126920 TI - Acute hyperhomocysteinemia induces a reduction in arterial distensibility and compliance. AB - OBJECTIVE: The aim of the study was to evaluate the effects of acute hyperhomocysteinemia on distensibility and compliance of large peripheral arteries. Isoprostanes generation and antioxidant vitamins were used to assess the role of oxidative stress. DESIGN: A cross-over, double-blind study on distensibility (DC: distensibility coefficient) and compliance (CC: cross sectional compliance) of common femoral and brachial arteries was performed in 12 healthy young male volunteers by means of a wall track system before and 4 h after a single oral methionine (100 mg/kg) or placebo administration. The effects of methionine load were investigated also after oral administration of vitamin C (1g/day) and vitamin E (800 mg/day) for 8 consecutive days. RESULTS: Oral methionine induced a significant increase in plasmatic levels of homocysteine. Distensibility and compliance of brachial and femoral arteries were significantly reduced after methionine load in comparison to placebo. This acute impairment of arterial wall mechanical properties was associated to endothelial dysfunction, since altered flow-dependent vasodilatation (P < 0.05 versus placebo) was observed in the same arterial districts. A significant increase in urinary 8-iso prostaglandin F2alpha was observed after methionine. Pretreatment with vitamins C and E prevented the effects of methionine on femoral and brachial arteries as well as on urinary 8-iso-prostaglandin F2alpha excretion. CONCLUSIONS: Hyperhomocysteinemia seems responsible for altered arterial wall elasticity and for endothelial dysfunction. A pivotal role can be attributed to oxidative stress. PMID- 15126921 TI - Abnormal regulation of G protein alpha(i2) subunit in skin fibroblasts from insulin-resistant hypertensive individuals. AB - BACKGROUND: Studies in experimental animals and human cells have demonstrated increased intracellular calcium (Ca(i2) signalling and Galphai signal transduction associated with hypertension. We have recently shown that angiotensin II-induced mobilization of Ca(i2) is enhanced in fibroblasts from hypertensive individuals in comparison with that in normotensive individuals and that it is blunted by insulin and pertussis toxin in insulin-sensitive, but not in insulin-resistant, patients. This suggests that G(i)-mediated signal transduction is reduced in insulin-resistant hypertension. OBJECTIVE: To investigate the expression and regulation of Galpha(i2) subunit in insulin sensitive and insulin-resistant hypertensive individuals. METHODS: G protein alpha(i2) subunit mRNA was measured in cultured skin fibroblasts from patients with insulin-sensitive and insulin-resistant hypertension, by real-time reverse transcriptase polymerase chain reaction. We also investigated the effects of short-term exposure to fetal calf serum, angiotensin II and insulin, alone and in combination, on the expression of Galpha(i2) in vitro. Spectrofluorophotometric measurement of free Cai was performed in monolayers of 24 h serum-deprived cells in basal conditions and after exposure to angiotensin II, with and without pre incubation with insulin. RESULTS: Expression of Galpha(i2) was significantly greater in fibroblasts from hypertensive individuals than in normotensive individuals and the increase was unrelated to age and body mass. The difference was largely accounted for by greater values in insulin-sensitive than in insulin resistant hypertensive individuals. In fibroblasts from those with insulin sensitive hypertension, angiotensin II and insulin were additive to fetal calf serum in increasing the expression of Galpha(i2). In these patients, insulin blunted the angiotensin-II induced Cai transient. In contrast, in those with insulin-resistant hypertension, Galpha(i2) was lower and unresponsive to angiotensin II and insulin. Finally, in fibroblasts from insulin-resistant patients, insulin was unable to reduce the angiotensin II-induced Cai peak. CONCLUSIONS: A subnormal Galpha(i2)-mediated signal transduction may be involved in the pathogenesis of cellular insulin resistance in hypertension. This novel Galpha(i2)-mediated signal transduction associated with insulin sensitivity in fibroblasts may help to control excessive angiotensin II signalling. PMID- 15126922 TI - Prevention of angiotensin II-induced hypertension, cardiovascular hypertrophy and oxidative stress by acetylsalicylic acid in rats. AB - BACKGROUND: Angiotensin II (Ang II)-induced oxidative stress has been suspected to play an important part in the pathogenesis of many cardiovascular diseases. Our previous study demonstrated that acetylsalicylic acid (ASA) possesses potent antioxidative properties. OBJECTIVE: To evaluate the pathogenetic role of oxidative stress in Ang II-induced hypertension and cardiovascular hypertrophy. METHODS AND RESULTS: Chronic infusion of Ang II (200 ng/kg per min for 12 days) increased the aortic and cardiac tissue production of superoxide anion (O2) (lucigenin-enhanced chemiluminescence method) by 77 and 35%, respectively. These effects were associated with progressive increases in systolic blood pressure (from 135 to 194 mmHg) and heart/body weight ratio (from 2.25 to 2.69). Chronic treatment with oral ASA alone (100 mg/kg per day for 12 days) significantly reduced aortic and cardiac production of O2 (by 31 and 33%, respectively), without alteration in blood pressure and heart/body weight ratio in control normotensive animals. However, concurrent treatment with ASA in Ang II-infused rats completely prevented the Ang II-induced production of O2, in addition to hypertension and cardiac hypertrophy. Similar protective effects were observed in cultured aortic smooth muscle cells, in which increases in O2 production and [H]leucine incorporation (221 and 38%, respectively) induced by Ang II (10 mol/l) were totally prevented by concurrent incubation with ASA (10 mol/l). Losartan, but not PD 123319, also blocked the Ang II-induced oxidative and hypertrophic effects in those cells. Other anti-inflammatory drugs, such as salicylic acid, indomethacin and ibuprofen, did not show similar anti-Ang II and antioxidative effects in vivo. CONCLUSIONS: Oxidative stress plays a major part in chronic Ang II-induced hypertension and cardiovascular hypertrophy. Chronic concurrent treatment with ASA was found to prevent those Ang II-induced effects on the cardiovascular system, presumably through its antioxidative properties. PMID- 15126923 TI - Hypertension, sodium retention, calcium excretion and osteopenia in Dahl rats. AB - BACKGROUND: Salt-sensitive hypertension in the Dahl rat is associated with abnormalities in both calcium (Ca2+) and sodium (Na) homeostasis. OBJECTIVE: To test the hypothesis that salt-induced abnormal Ca(2+) handling in Dahl salt sensitive (DSS) rats is associated with negative Ca(2+) balance and bone disease. METHODS: Ca(2+) excretion in acute and chronic Na(+) loading and electrolyte and water balance were determined by balance studies in Dahl salt-resistant (DSR) and salt-sensitive (DSS) rats fed 8 or 0.1% NaCl for 4 weeks. A dry ashing procedure was used to determine Na(+), Ca(2+), and water content and their association with blood pressure in the rats. RESULTS: When fed 8% NaCl, DSS rats initially maintained a positive Ca(2+) balance and showed decreased natriuresis compared with DSR rats. During the course of Na(+) loading, DSS rats increased natriuresis and calciuresis. After 4 weeks of salt loading, cumulative Na balance was greater and cumulative Ca(2+) balance was less in DSS than in DSR rats. In addition, DSS rats developed osteopenia. Bone mineral content correlated inversely with blood pressure in DSS rats. Acute saline volume expansion in DSS rats demonstrated their ability to excrete the Na load fully, but led to an exaggerated renal loss of Ca(2+) compared with DSR rats. CONCLUSION: DSS, but not DSR, develop Ca(2+) loss and ostopenia during chronic Na(+) loading. We speculate that Na retention in DSS rats fed a high Na diet may be in part a compensatory mechanism to maintain Ca(2+) balance. PMID- 15126924 TI - Elevated arterial pressure impairs autoregulation independently of AT(1) receptor activation. AB - OBJECTIVE: These studies determined the ability of AT1 receptor blockade or 'triple therapy', to reverse angiotensin II-induced hypertension and improve autoregulatory behavior. DESIGN: Experiments to determine if regulation of systolic blood pressure, in the normotensive range, would improve renal microvascular autoregulatory behavior in angiotensin II-infused rats. METHODS: Hypertension was induced by chronic angiotensin II infusion (60 ng/min) for 10-14 days. Two groups of angiotensin II-infused rats received either AT1 receptor blockade, with candesartan cilexetil, or triple therapy, with hydralazine, hydrochlorothiazide and reserpine, beginning on day 6 or day 0 of angiotensin II infusion, respectively. Sham animals were studied as normotensive controls. Systolic blood pressure was measured by tail cuff. Autoregulatory behavior was assessed using the juxtamedullary nephron technique in response to step (15 mmHg) increases in perfusion pressure from 65 to 170 mmHg. RESULTS: Angiotensin II infusion increased systolic blood pressure from a baseline of 125 mmHg to 162 and 182 mmHg after 10 and 14 days, respectively. Candesartan cilexetil and triple therapy normalized the blood pressure to between 119 and 126 mmHg. Increasing perfusion pressure, from 65 to 170 mmHg, reduced afferent arteriolar diameter by 30% in sham-treated kidneys. Autoregulation was significantly blunted in angiotensin II-infused rats, resulting in a pressure-mediated vasoconstriction of only 10%. Candesartan cilexetil, or triple therapy, significantly improved autoregulatory behavior, as indicated by pressure-mediated vasoconstrictor responses of 30 and 40%; respectively, despite continued angiotensin II infusion. CONCLUSIONS: These data suggest that chronic elevation of arterial blood pressure, rather than chronic AT1 receptor stimulation, is sufficient to induce hypertensive impairment of renal autoregulatory capability. PMID- 15126925 TI - Plasma and kidney angiotensin II levels and renal functional responses to AT(1) receptor blockade in hypertensive Ren-2 transgenic rats. AB - OBJECTIVE: The first aim of the present study was to assess plasma and kidney angiotensin II (ANG II) levels and renal cortical ANG II receptor subtype 1A (AT1A) mRNA expression in hypertensive Ren-2 transgenic rats (TGR) and in normotensive Hannover Sprague-Dawley (HanSD) rats. The second aim was to investigate potential differences between TGR and HanSD in blood pressure (BP) and renal functional responses to either intravenous (i.v.), i.e. systemic, or intrarenal (i.r.) AT1 receptor blockade with candesartan. METHODS: Rats were anesthetized and prepared for clearance experiments. In series 1, ANG II concentrations were assayed by radioimmunoassay and renal cortical AT1A mRNA expression by semiquantitative reverse transcriptase-polyacrylamide gel electrophoresis. In series 2, BP and renal functional responses were evaluated after either i.v. or i.r. bolus administration of candesartan. RESULTS: Plasma and kidney ANG II levels were significantly lower in TGR than in HanSD (39 +/- 5 versus 107 +/- 19 fmol/ml and 251 +/- 41 versus 571 +/- 95 fmol/g, respectively, P < 0.05). Renal AT1A mRNA expression was not different between TGR and HanSD. Intravenous candesartan caused comparable decreases in BP in TGR and HanSD and did not change renal plasma flow (RPF) or absolute and fractional sodium excretion in HanSD. In contrast, i.v. candesartan significantly increased RPF (+27 +/- 6%, P < 0.05) and absolute and fractional sodium excretion (+49 +/- 10 and + 42 +/- 9%, respectively P < 0.05) in TGR without changing glomerular filtration rate (GFR). Acute i.r. candesartan increased RPF by +36 +/- 6% (P < 0.05) in TGR but not in HanSD with a greater rise in absolute and fractional sodium excretion in TGR (+124 +/-8 and 97 +/- 9%, respectively) than in HanSD (+81 +/- 9 and +69 +/- 8%, respectively) (P < 0.05). CONCLUSIONS: The enhanced responses of RPF and sodium excretion to AT1 receptor blockade in TGR suggest that renal hemodynamics and sodium excretion in TGR are under strong ANG II influence. The compromised ability of the kidney to respond to BP elevations by appropriate increases in sodium excretion may contribute to the maintenance of high BP in TGR. Thus, the present findings provide new insights into the pathophysiology of hypertension in this model. PMID- 15126926 TI - Amlodipine at high dose increases preproendothelin-1 expression in the ventricles and aorta of normotensive rats. AB - BACKGROUND: High doses of dihydropyridine calcium channel blockers can activate the sympathetic nervous system and the renin-angiotensin system. Both noradrenaline and angiotensin II stimulate preproendothelin-1 gene expression, yet the effects of high doses of dihydropyridines on preproendothelin-1 expression in vivo remain unknown. OBJECTIVES: To investigate the effects of high doses of dihydropyridines on preproendothelin-1 expression in the ventricles and aorta of normotensive rats. METHODS: Sprague-Dawley rats were treated with amlodipine 5 or 20 mg/kg per day (Amlo 5 or Amlo 20) in drinking water for 5 days or 5 weeks. Systolic blood pressure and heart rate were measured by tail-cuff plethysmography. Gene expression was examined by reverse transcriptase polymerase chain reaction. RESULTS: Amlo 5 increased heart rate during the first week only and had no effect on blood pressure and ventricular weight and gene expression. Amlo 20 reduced blood pressure transiently and increased heart rate consistently. It did not change relative left ventricular weight (corrected for body weight) after 5 days, but increased it after 5 weeks; it increased relative right ventricular weight at both time points. Aorta weight (mg/mm) was decreased after 5 weeks of treatment with both dosages of amlodipine. Preproendothelin-1 mRNA levels were increased by Amlo 20 in the ventricles and aorta and, concomitantly, renin mRNA was increased in the kidney. Less consistently, interleukin-6 mRNA also increased in ventricles, whereas cardiotrophin-1 mRNA remained unchanged. The sensitivity of isolated aorta to the contractile effect of noradrenaline was decreased by Amlo 5, but not by Amlo 20. CONCLUSIONS: In Sprague-Dawley rats, high-dose amlodipine, while promoting neurohormonal activation, induced overexpression of preproendothelin-1 mRNA in the ventricles and aorta. Endothelin 1 overexpression could contribute to the lack of inhibitory effect of high-dose amlodipine on ventricular mass in normotensive rats. PMID- 15126927 TI - Effects of telmisartan 80 mg and valsartan 160 mg on ambulatory blood pressure in patients with essential hypertension. AB - OBJECTIVES: This trial investigated and compared the antihypertensive efficacy of telmisartan and valsartan, two angiotensin II receptor blockers, used in monotherapy at their maximum recommended dose in hypertensive patients. METHODS: We studied 70 subjects (32 men and 38 women) aged 47.6 +/- 12.2 (mean +/- SD) years, with mild to moderate essential hypertension; they were randomly assigned to receive monotherapy with either telmisartan (80 mg) or valsartan (160 mg), in the form of a single daily tablet upon awakening. Blood pressure was measured by ambulatory monitoring every 20 min during the day and every 30 min at night for 48 consecutive hours before and after 3 months of treatment. Physical activity was simultaneously monitored every minute by wrist actigraphy to calculate accurately the diurnal and nocturnal means of blood pressure on a per subject basis. RESULTS: There was a highly significant blood pressure reduction during the 24 h with both drugs. The blood pressure reduction in the 24-h mean was significantly larger for valsartan 160 mg (18.6 and 12.1 mmHg for systolic and diastolic blood pressure, respectively) than for telmisartan 80 mg (10.8 and 8.4 mmHg; P < 0.001 between treatment-groups). There was also a highly significant reduction (P < 0.001) of 6.5 mmHg in the 24-h mean of pulse pressure after valsartan administration only. The trough : peak ratio and the smoothness index were slightly higher in systolic, but similar in diastolic blood pressure, for telmisartan as compared to valsartan. CONCLUSIONS: Despite a shorter half-life, 160 mg/day valsartan was more effective in lowering blood pressure over 24 h than 80 mg/day telmisartan. Furthermore, valsartan was also more effective in lowering arterial pulse pressure, an observation that may have important therapeutic implications, given the mounting evidence that pulse pressure may be a risk factor for future cardiovascular events. PMID- 15126928 TI - Effects of immediate versus delayed antihypertensive therapy on outcome in the Systolic Hypertension in Europe Trial. AB - BACKGROUND: To assess the impact of immediate versus delayed antihypertensive treatment on the outcome of older patients with isolated systolic hypertension, we extended the double-blind placebo-controlled Systolic Hypertension in Europe (Syst-Eur) trial by an open-label follow-up study lasting 4 years. METHODS: The Syst-Eur trial included 4695 randomized patients with minimum age of 60 years and an untreated blood pressure of 160-219 mmHg systolic and below 95 mmHg diastolic. The double-blind trial ended after a median follow-up of 2.0 years (range 1-97 months). Of 4409 patients still alive, 3517 received open-label treatment consisting of nitrendipine (10-40 mg daily) with the possible addition of enalapril (5-20 mg daily), hydrochlorothiazide (12.5-25 mg daily), or both add-on drugs. Non-participants (n = 892) were also followed up. RESULTS: Median follow up increased to 6.1 years. Systolic pressure decreased to below 150 mmHg (target level) in 2628 participants (75.0%). During the 4-year open-label follow-up, stroke and cardiovascular complications occurred at similar frequencies in patients formerly randomized to placebo and those continuing active treatment. These rates were similar to those previously observed in the active-treatment group during the double-blind trial. Considering the total follow-up of 4695 randomized patients, immediate compared with delayed antihypertensive treatment reduced the occurrence of stroke and cardiovascular complications by 28% (P = 0.01) and 15% (P = 0.03), respectively, with a similar tendency for total mortality (13%, P = 0.09). In 492 diabetic patients, the corresponding estimates of long-term benefit (P < 0.02) were 60, 51 and 38%, respectively. CONCLUSIONS: Antihypertensive treatment can achieve blood pressure control in most older patients with isolated systolic hypertension. Immediate compared with delayed treatment prevented 17 strokes or 25 major cardiovascular events per 1000 patients followed up for 6 years. These findings underscore the necessity of early treatment of isolated systolic hypertension. PMID- 15126929 TI - ESH/ESC guidelines. PMID- 15126931 TI - Aldosterone excess in hypertension. PMID- 15126933 TI - Exercise-induced asthma: ways to wise exercise. PMID- 15126934 TI - The role of hyposensitization: do we need to start rethinking? PMID- 15126935 TI - Mediators of inflammation in the early and the late phase of allergic rhinitis. AB - PURPOSE OF REVIEW: This review provides the reader with an overview of the early and the late phase of the immediate allergic reaction. Furthermore, recent publications are presented that could reveal important new insights into the mechanisms of allergic inflammation. RECENT FINDINGS: Recent work on the possible roles of nerve growth factor, nitric oxide synthases, several CC chemokines and other mediators is presented. SUMMARY: Allergic rhinitis is characterized by the clinical symptoms of sneezing, itching, rhinorrhea, and nasal congestion. Frequently, these are accompanied by eye, ear and throat symptoms or postnasal drip. The symptoms arise as a result of inflammation induced by IgE-mediated immune response to a specific allergen. Inflammatory mediators are released and cells are activated and recruited to the mucosa. In this review, early and late phase responses of the allergic type I reaction are described, including the different cell types and mediators involved. Special attention is paid to new inflammatory processes to provide the reader with recent information. PMID- 15126936 TI - Control of nasal obstruction in perennial allergic rhinitis. AB - PURPOSE OF REVIEW: Nasal obstruction, the cardinal symptom of persistent (perennial) allergic rhinitis, is one of the most common symptoms encountered in primary care and in specialist clinics. It is difficult to quantify by clinical examination, and, hence, objective assessment of the nasal airway is critical to rhinologic research. Nasal obstruction in persistent allergic rhinitis must be treated the year round, and therefore treatment choices, costs, and compliance all become important public health issues. RECENT FINDINGS: Many inflammatory and neurogenic mediators released during allergic reactions are able to cause plasma exudation and vasodilatation, with resultant edema and swelling of the nasal mucosa. Recently, technological advancements have made it possible to qualitatively and quantitatively study the nasal airway, providing greater insights into the understanding of physiological fluctuation and pathophysiological manifestations of nasal patency. From recent international guidelines, the management of allergic rhinitis includes combining treatments of the upper and lower airways, by using patient education, allergen avoidance, pharmacological treatment, and specific immunotherapy. Surgery may be needed as an adjunctive intervention. Multiple methods have been introduced to treat turbinate hypertrophy. However, preservation of adequate nasal mucosal function is important, together with long-term results. SUMMARY: It is important that consensus recommendations for the management of allergic rhinitis be designed and implemented by all levels of medical specialists in order to improve treatment outcomes. PMID- 15126937 TI - Quality of life scales in allergic rhinitis. AB - PURPOSE OF REVIEW: The recent literature on types, development, validation, indications, and limitations of quality of life scales in allergic rhinitis is reviewed. This review does not report clinical trials that included quality of life as an outcome. RECENT FINDINGS: Today, quality of life measurement is standard in allergic rhinitis. Different disease-specific and generic questionnaires are very well validated and established. Recent research concerns the development or adaptation of questionnaires for specific purposes or populations. Other research focuses on the registration of concomitant illnesses, especially of asthma, taking the model of rhinitis and asthma as 'one airway disease' into account. Recently, the first questionnaire for rhinitis and asthma was presented. Furthermore, attempts have been made to improve the registration of other local or systemic concomitant illnesses, the sensitivity to rapid changes in the patient's condition, and the objectivity of quality of life scales. SUMMARY: Allergic rhinitis is associated with significant impairments of quality of life. Disease-specific quality of life scales offer a better registration of changes in disease-related problems, whereas generic scales allow for a comparison between different disorders and patient populations. Different versions of quality of life scales have been validated for different purposes. Careful selection of quality of life scales is becoming more and more important. The registration of comorbid conditions and of psychological factors influencing quality of life measurement remains a central problem. For research, the combination of disease-specific and generic quality of life scales may offer new perspectives; for clinical use, patient-friendly scales should be used. PMID- 15126938 TI - An update on the asthma-rhinitis link. AB - PURPOSE OF REVIEW: For the present article we collected and reviewed the more relevant experimental results concerning the asthma-rhinitis link in allergic diseases, published since January 2002. RECENT FINDINGS: During the last 2 years, particular attention has been devoted to the behaviour of the immune response in the two compartments of the airways. The recent experimental data, mainly obtained with specific nasal or bronchial allergen challenges, have confirmed that the link between the nose and the bronchi is bidirectional, and that a systemic cross-talk occurs. Furthermore, the pathogenic role of paranasal sinus infections in respiratory allergy has been better elucidated. It was shown that, in sinusitis, a T helper type 2 polarization exists, which can be reverted by proper therapy. On the other hand, despite the abundant experimental evidence, our view of the united airways is still not complete, and several points need to be developed. SUMMARY: The new findings on the asthma-rhinitis link have confirmed the current pathogenic view of respiratory allergy. These findings have important implications from a therapeutic point of view, and therefore encourage and promote the search for novel integrated treatment strategies. PMID- 15126939 TI - Gaining insight into patients' beliefs using qualitative research methodologies. AB - PURPOSE OF THE REVIEW: Qualitative research is a rigorous inductive approach to data collection and data interpretation used to describe patients' perspectives and behaviors. Qualitative research can also be used to develop hypotheses to be tested with quantitative procedures. Combining qualitative and quantitative methodologies can provide more-comprehensive explanations for health actions and beliefs than can be derived from either approach alone. This review serves as a primer for those health care professionals who are unfamiliar with this alternative research paradigm. RECENT FINDINGS: We will explore the similarities and differences between quantitative and qualitative techniques and describe how focus groups allowed us an enhanced appreciation of the reasons for poor inhaled corticosteroid adherence in low-income African-American patients with persistent asthma. SUMMARY: As populations become more diverse, providers are under increased pressure to effectively communicate and partner with their patients. Qualitative research offers the practitioner a roadmap for enhanced understanding of the unique experiences of patients, thus promoting quality patient-provider relationships. PMID- 15126940 TI - Medication non-adherence and asthma treatment cost. AB - PURPOSE OF REVIEW: The purpose of this review is to describe the impact of asthma treatment non-adherence on patients and the healthcare system, and to outline areas of responsibility towards improved adherence. RECENT FINDINGS: The average cost of healthcare expenses for each person in the United States in 2002 was 5440 US dollars. In that year, there were 800 million medical encounters. However, adherence research suggests that a significant portion of the healthcare advice and prescriptions dispensed in these encounters was wasted. The annual cost to the healthcare system caused by non-adherence has been estimated at 300 billion US dollars. The responsibility for improving adherence has usually been placed on the patient and healthcare provider. However, if non-adherence is to be more effectively addressed, other components of the healthcare and pharmaceutical industries must also take responsibility. SUMMARY: Treatment non-adherence compromises treatment effectiveness and drives up the healthcare costs related to asthma and other chronic conditions. Collaborative efforts to improve adherence to treatments for chronic illness, recently promoted by the World Health Organization, must include multiple components of the healthcare system, must recognize that the costs of adherence promotion are outweighed by cost savings after improved adherence, and must support research to develop new and better strategies for improving adherence. PMID- 15126941 TI - The mechanism of food allergy: what do we know today? PMID- 15126942 TI - Peanut, tree nut and seed allergies. PMID- 15126943 TI - Food allergy: what do we learn from animal models? AB - PURPOSE OF REVIEW: This review summarizes selected articles on animal models of food allergy published in 2003. The research areas that are covered include mechanistic studies, the search for new therapies, as well as screening models for hazard identification of potential allergens. RECENT FINDINGS: Novel treatment options of both prevention and therapeutic strategies have been reported with promising results. The induction of de-sensitization to food proteins was achieved by exposure to a mixture of recombinant food allergens and T helper 1 (Th1)-skewing bacterial components. Furthermore, research in animal models has provided new insights into the role of protein structure, digestion, and gut permeability in sensitization and tolerance induction to food proteins. The Th2 hypothesis of food allergy was tested in mouse strains, linking genetic susceptibility to sensitization with differential Th1-Th2 responses. In this context, the role of the liver in development of food antigen-specific Th2 cells, and the importance of costimulatory molecules in Th2 skewing were demonstrated. Finally, rodent models to predict potential allergenicity of novel foods have been further developed using different routes of sensitization. SUMMARY: Currently, several animal models of food allergy are used, including mouse, rat, swine, and dog. Continuing research in these models may elucidate the immunological mechanisms that underlie the sensitization and challenge phase of food allergy and may result in improved therapeutic options. Furthermore, the development of animal models to predict relative allergenicity of novel foods remains an important topic. PMID- 15126944 TI - Do we have suitable in-vitro diagnostic tests for the diagnosis of food allergy? AB - PURPOSE OF REVIEW: Positive standardized food challenges represent the gold standard of diagnostic procedures in food-related reactions suspected to be of allergic nature. Skin prick testing and in-vitro diagnosis is helpful in most cases and can help to avoid cumbersome food challenges. This review considers recent progress in the use of in-vitro tests in the diagnosis of food allergy. RECENT FINDINGS: Recent studies have addressed the characterization of 'new' food allergens which might now be used more accurately in the in-vitro diagnosis of food allergy. Additionally, while in-vitro tests must always be interpreted in line with the allergen tested and the clinical history, levels of food-specific immunoglobulin E can be correlated with the outcome of challenges to foods such as tree nuts as well as egg, according to two recent studies. Finally, epitope binding patterns of specific food allergens might help to predict which patients will most likely outgrow their food allergy, or which patients are clinically tolerant. This might help to avoid food challenges, which carry a risk for a potentially severe outcome. SUMMARY: Recent studies of in-vitro diagnosis of food allergy have helped to provide safer and more accurate tests in the diagnosis and prognosis of food allergy. PMID- 15126945 TI - Update on threshold doses of food allergens: implications for patients and the food industry. AB - PURPOSE OF REVIEW: The purpose of this review is to bring the reader up to date on the importance of assessing a food's lowest observed adverse-effect level (LOAEL) with two aims. Firstly, to help industry choose tests with a level of sensitivity capable of detecting food allergens hidden in industrial products. Secondly, to specify protective measures for highly allergic individuals in order to prevent recurrent severe anaphylaxis. The review also seeks to highlight the present issues and unsolved questions. RECENT FINDINGS: Thanks to standardized oral-provocation tests (double-blind placebo-controlled food challenges), LOAELs have been identified for many IgE-dependent food allergies. Most studies concern the pediatric population. Data is available for milk, egg, peanut, wheat flour, and sesame. The LOAELs are commonly in the range of 1-2 mg of natural foods, representing a few hundred micrograms of protein. These minimal reactive doses characterize about 1% of people allergic to milk, egg, or peanut. The level at which no observed adverse effect is seen might be a few tens of micrograms of protein for peanut. At the present time, allergy to oil seems to be restricted to unrefined cold-pressed oils. SUMMARY: Concerning IgE-dependent food allergies, the threshold dose inducing symptoms is now known to vary a great deal according to the individual. A reactive dose of less than 65 mg characterizes 16 and 18% of patients allergic to egg or peanut. Less than 30 mg of milk proteins characterizes 5% of those allergic to milk. For milk, egg, and peanut, 1% of patients have a very low threshold, about 1 mg. Such data emphasize the necessity of using detection tests with a sensitivity better than 10 parts per million. The modifications of allergenicity undergone by protein ingredients that are now commonly introduced into industrially made products are not yet sufficiently known. A better knowledge of the reactive doses of these proteins is needed. PMID- 15126946 TI - Pathophysiology, diagnosis and treatment of food protein-induced gastrointestinal diseases. AB - PURPOSE OF REVIEW: Although our general understanding of food hypersensitivity has improved in recent years, gastrointestinal food protein-induced diseases still pose diagnostic and therapeutic dilemmas. RECENT FINDINGS: Food allergy in children and adults may involve any part of the gastrointestinal tract. Clinical presentations include protein-induced enterocolitis syndrome, enteropathy and proctocolitis, as well as eosinophilic gastroenteritis and related disorders. For many of these conditions, our understanding of the pathophysiology is incomplete. Manifestations are mostly non-IgE mediated, and skin prick testing and measurement of food-specific IgE antibody levels are of limited diagnostic value. Atopy patch testing may be of benefit in identifying food items associated with late-onset gastrointestinal reactions. A definitive diagnosis of gastrointestinal food allergy, however, still relies on formal food challenges. Depending on the clinical presentation, gastrointestinal biopsies may be required. In infancy, hypoallergenic formula or maternal elimination diets have been shown to effectively control the gastrointestinal manifestations of food allergies. Growth parameters and micronutrient levels need to be carefully monitored while on elimination diets for prolonged periods. In older children and adults with eosinophilic gastrointestinal disorders, the response to dietary restriction is variable. Corticosteroids may be required to control symptoms in those who failed to respond to hypoallergenic diets. In eosinophilic esophagitis, steroids can be administered topically in the form of swallowed aerosols. Leukotriene receptor antagonists and other novel therapies may be useful as steroid-sparing agents. SUMMARY: Early diagnosis and treatment of food protein-induced gastrointestinal diseases may prevent significant nutritional complications. Further research is needed to develop diagnostic tools for these mainly cell-mediated disorders. PMID- 15126947 TI - Seafood allergy. AB - PURPOSE OF REVIEW: Given the importance and prevalence of seafood in our economy and diet, interest in the adverse effects of seafood-induced allergic reactions has increased both in the lay population and the scientific arena. The purpose for this review article is to provide an overview and discussion of current seafood-allergy research. RECENT FINDINGS: Current research has produced the discovery of new seafood allergens as well as a better understanding of the interactions of allergens with the host, and the relationships of IgE antibodies with specific allergens. Crossreactivity of similar molecules from seafood and nonseafood sources provides a better understanding of an allergen's role in seafood allergy and a basis to improve patient identification and treatment. SUMMARY: Findings in recent literature show potential for more precise diagnosis and safer, more effective treatment of seafood allergies. PMID- 15126948 TI - Clinical importance of cross-reactivity in food allergy. AB - PURPOSE OF REVIEW: Review of recent developments in the field of cross-reactivity in food allergy and the clinical relevance of these developments. RECENT FINDINGS: New foods have been added to the list of Bet v 1 and profilin-related food allergies. Clinical relevance of cross-reactions based on recognition of carbohydrate determinants and profilin is limited for the population of pollen allergic patients as a whole. For selected food allergic patients, however, N glycans and particularly profilin are potentially of clinical relevance. Lipid transfer proteins have further been established as clinically more severe allergens in several foods. This severity is attributed to their stability to proteolysis and processing. Storage proteins of several nuts and seeds have been identified as important allergens, but cross-reactivity between storage proteins of different foods appears to be limited. Using cross-reactivity as the basis for immunotherapy in food allergy seems promising but needs confirmation by double blind, placebo-controlled trials. SUMMARY: The continued identification and characterization of cross-reactive allergens facilitates the study of factors determining clinical relevance of cross-reactivity and of possible efficacy of immunotherapy in food allergy. PMID- 15126949 TI - Food processing: effects on allergenicity. AB - PURPOSE OF REVIEW: There has been an increase in the prevalence of food allergy, asthma and severe reactions to foods in the past decade. The reason for this increase is unknown and despite the potential for a lethal outcome, no treatments or therapies are available. Many areas are being explored to understand the increase in food allergy and the role of modern processing techniques is one of them. RECENT FINDINGS: The fact that processing influences allergenicity is not a new concept, but due to some definitive studies in the last few years the topic has been revitalized. Not only is the processed form of a particular allergen important in assessment of allergenicity, but the role of the food matrix in which an allergen is processed must also be evaluated. The issue of allergenicity of particular foods is therefore highly complex, and even with our existing knowledge we have just begun to understand some of the contributions of processing to allergenic properties of proteins and foods. SUMMARY: Researchers and medical personnel have begun to weigh the benefits and potential disadvantages of assessing the allergenicity of foods in unprocessed forms. In this review, the known effects of processing on the allergenicity of various foods will be discussed along with the implications of the most recent studies on the future direction of research in this area. PMID- 15126950 TI - Bibliography. Current world literature. Outcome measures. PMID- 15126952 TI - Standardize or individualize: finding a balance. PMID- 15126953 TI - The experiences of district nurses caring for people receiving palliative chemotherapy. AB - The treatment options being offered to people with advanced cancer are increasing with growing use of palliative chemotherapy. As people are experiencing shorter hospital stays and receiving treatment on an outpatient basis, this has implications for primary health-care provision. This study aimed to explore the experiences of district nurses caring for patients receiving palliative chemotherapy: how they viewed their role, factors that influenced their role and their attitudes to palliative chemotherapy. Data were collected using qualitative interviews that incorporated critical incident technique (n=10). Themes that emerged from the content analysis included the role of the district nurse, knowing the patient and family, the interface between hospital and primary care, and uncertain ground. District nurses saw their role as having relevance at all stages of the patient's cancer journey and the provision of holistic care based on good interpersonal relationships was valued. However, difficulties were perceived at the interface between hospital and primary care. District nurses had ambivalent attitudes to palliative chemotherapy but had positive attitudes towards optimizing quality of life and care in the palliative stages of illness. PMID- 15126954 TI - Promoting ownership in palliative care audit. PMID- 15126955 TI - NICE supportive and palliative care guidance. PMID- 15126956 TI - Discharging patients from hospice to nursing home: a retrospective case note review. AB - This article presents a retrospective case note review of in-patients at one UK hospice considered for discharge to nursing homes during 1999 and 2000. The idea of a nursing home placement was introduced, on average, around 9 days after admission to the hospice; the most common reason cited being the inability to cope at home. Nearly a third of the patients expressed negative or ambivalent feelings about the idea. Transferring patients from hospice to nursing home is often necessary to avoid pressures on hospice beds. The potential distress of transferring patients, particularly when nearing the end of life, must be balanced against the needs of patients on hospice waiting lists. Of the 41 cases reviewed, 16 were transferred to nursing homes as planned, with a mean length of stay of just over 8 weeks. The current provision of palliative care in nursing homes raises ethical questions about transferring patients to an unfamiliar environment that may not necessarily be able to offer the same quality of palliative care. Clear communication with patients and their families and a consistent process are important to reduce some of the distress associated with this issue. PMID- 15126957 TI - Spiritual distress in a terminally ill patient with breast cancer. PMID- 15126958 TI - The experience of family members caring for a dying loved one. AB - Qualitative nursing research into the experience of family members caring for a dying loved one has been limited. This study used a phenomenological approach to explore this experience. The pattern of caring for a dying loved one and its intertwined dimensions were described. Caregivers felt a sense of helplessness that was associated with illness progression, their inability to relieve pain and discomfort, and decision-making related to patient admission to a palliative care unit. Lack of support from health professionals and having to face personal limits were found to accelerate the decision to admit a patient. The role of a support person involved with the caregiver was also considered and found to be an area worthy of further investigation. Health professionals must provide information and support tailored to the caregivers' needs as they change along a patient's illness trajectory. PMID- 15126960 TI - Government survey finds that NHS staff are generally satisfied. PMID- 15126959 TI - Home care versus hospital care in patients with multiple myeloma treated with pamidronate. AB - Long-term bisphosphonate therapy has been shown to offer clinical benefit in the management of multiple myeloma. This study sought to explore the feasibility and potential advantages of monthly home-based intravenous infusions of pamidronate in patients with multiple myeloma. In a prospective crossover, multicentre trial, 37 patients were randomly allocated to receive 3 months of treatment with pamidronate given in the home followed by 3 months of treatment with pamidronate given in hospital or vice versa. Results from a patient preference questionnaire indicated most patients preferred treatment at home. Quality-of-life measurement was undertaken using the EORTC QLQ-C30 questionnaire. The results indicated a small, generally consistent, although not statistically significant, trend in favour of home care treatment. Extra nursing specialist time was required for home therapy. Home therapy with pamidronate in patients with multiple myeloma appeared feasible and safe and was preferred by patients in this study. PMID- 15126961 TI - Preserving and enhancing standards of care in the NHS. PMID- 15126962 TI - Nurse who took on responsibilities beyond her competence. PMID- 15126963 TI - A review of osteoporosis in men: implications for practice. AB - This article discusses the issue of osteoporosis from a male perspective. Osteoporosis is often seen as a woman's disease; however, there are a significant number of men who suffer from osteoporosis and the effects of the disease can have implications for the man's physical, social and psychological well-being. The reasons why osteoporosis may be increasing in the male population are discussed. The possible causes of osteoporosis as well as the risk factors that may put men at danger of contracting the illness are outlined. Bone density is an important factor related to the development of osteoporotic fractures - methods of measuring bone density using dual energy X-ray absorptiometry are provided, as are other important investigations that will be needed. The management and treatment of the male patient with osteoporosis and the role of the nurse are detailed. PMID- 15126964 TI - The management and care of people with stoma complications. AB - There are a variety of complications that are associated with stomas. These are described and their causes explained. A number of solutions are presented and advice that the nurse can give is offered. Some of the complications are commonly seen by the nurse, such as sore skin, whereas others are rare, such as an allergy to the stoma appliance. There may be sexual dysfunction after surgery, which may be temporary or permanent. These and many other potential problems are elaborated upon to aid the nurse when caring for people with stomas. PMID- 15126965 TI - Living with sickle cell disease: the perspective of young people. AB - The study aimed to explore the lived experience of young people with sickle cell disease as they transferred to adult services. A cross-sectional study using semistructured questionnaires is described. Eleven young people (12-16 years) attending a sickle cell and thalassaemia centre as they approached transition from paediatric to adult services were successfully recruited to the study. Sickle cell disease was reported to interfere with various aspects of their 'normal' lives. Fatigue and pain were common symptoms. Reported adherence to prophylaxis was variable. It was concluded that young people with sickle cell disease require support drawing on a range of expertise and that the medical model of service delivery may not meet all their needs. PMID- 15126966 TI - The views of children and their families on being in hospital. AB - This study aimed to elicit the separate views of children, young people and carers after a period in hospital as an inpatient. Questionnaires were administered to 130 children and their families discharged after a period as hospital inpatients in January 2003. Anonymized data were returned from 50 families. Data analysis indicated that there were differences in the way that the three groups perceived the period of admission. Although parental sleeping and other social arrangements were subject to some critical review, the nursing care experienced by families was highly rated. Although arrangements for discharge were deemed satisfactory, 38% of carers had to wait for medicines to arrive on the ward before they could go home. Only six of the young people felt there ward catered for their age group and five indicated poor levels of privacy. None of the young people indicated that they had used the equipped teenagers' room. Attempts to include the voice of the younger child in this study proved unsatisfactory as parents elected to act as proxies in completing the child specific questionnaires. Child healthcare professionals attempting to involve all service users in determining optimum levels of care need to consider fully the methods of data collection and their applicability for differing age groups of children. Dependence on adult carers to reflect accurately the voice of the child is not fully satisfactory. PMID- 15126967 TI - Accountability and medicinal products 4: professional issues. PMID- 15126968 TI - Basic nursing principles need to be remembered. PMID- 15126969 TI - We must act to move tissue viability forward. PMID- 15126970 TI - Specialists must accept challenge of improving clinical outcomes. PMID- 15126971 TI - A review of the use of silver in wound care: facts and fallacies. AB - This review traces the use of silver in wound care, discussing its merits as an antibacterial agent and constituent of many new dressings, which are increasingly tailored to the treatment of wounds ranging from acute surgical lesions to chronic and diabetic leg ulcers. Misconceptions regarding the biological properties of silver, its possible physiological value in the human body and wound bed, absorption through the skin, and safety factors are addressed. The article aims to present silver and the new range of sustained silver-release dressings as important features in the management of skin wounds, providing effective control of wound infections while ensuring patient comfort and quality of life. PMID- 15126972 TI - Evaluation of Urgotol plus K-Four compression for venous leg ulcers. AB - In this non-comparative clinical evaluation, 36 subjects with venous leg ulcers, 85% of which were indolent or deteriorating, were treated with Urgotul lipidocolloid wound dressing and the K-Four multilayer compression bandaging system for 12 weeks or to healing--whichever occurred first. Results show that Urgotul was an ideal dressing in combination with K-Four, being easy to apply (98.7%) and remove (98.1%), and largely pain-free (95.6%) and non-adherent (99.7%). In a patient group of "hard-to-heal" ulcers, 50% of the ulcers healed within the treatment period. Ulcers not healed after 12 weeks achieved almost 50% area reduction on average. The treatment combination proved safe, with only one of seven adverse events reported being probably related to the products used. This study supports the use of a combination of Urgotul dressing and K-Four compression to provide a "matched" treatment for venous leg ulcers. PMID- 15126973 TI - Hydrocapillary dressing to manage exudate in venous leg ulcers. AB - This study evaluated the safety and clinical performance of Alione hydrocapillary adhesive dressing, Coloplast A/S, a newly developed moist wound healing dressing designed for exudate management of heavily to lightly exuding ulcers. Twelve patients with heavily to moderately exuding chronic venous leg ulcers were assessed during a 4-week observation period. Assessments included healing progress, pain, odour, peri-ulcer skin condition, ease of use, patient comfort and suitability of the dressing for managing exudate. The median ulcer area was reduced by 71% (P<0.05) and the wound bed tissue improved markedly. No leakages of exudate were reported and the overall impression of the dressing was good. The amount of exudate assessed subjectively (P<0.05). Pain and odour were reduced effectively. No dressing-related adverse events were observed. This study has demonstrated that Alione is safe and easy to use, and exhibits convincing clinical performance and exudate-handling properties in the treatment of chronic venous leg ulcers. PMID- 15126974 TI - Comparison of four different dressings on donor site wounds. AB - Following nursing observation of the difficulties of healing of donor site wounds, including trauma and pain on removal of dressings, a blind randomized study was undertaken to determine which of four dressings provided the greatest comfort, had the fastest healing time and could be removed without trauma. Forty patients had one of four dressings applied in theatre: paraffin gauze, alginate alone, alginate with film cover or film alone. An observer was present who was unaware of the dressing used and recorded patient observations and nursing records for information. The study revealed that the dressing that caused patients most discomfort and trauma on removal was paraffin gauze. Alginate with film dressing was found to be most successful in treating these patients. PMID- 15126975 TI - The Plymouth skin and wound care service goes stateside. AB - In 2002 the authors were successful in bidding for an Innovations in Healthcare Travel Award donated through the British Journal of Nursing, in association with J&J Medical. With this grant they were able to travel to the USA and study health care in Boston Massachusetts. The trip was not without problems, both in arranging and subsequently visiting the area. However, both authors have learnt from the experience and have returned with lessons that can be implemented in their healthcare setting. The following is a personal account of that trip and the lessons learnt. PMID- 15126976 TI - Russka pressure-relieving low air-loss mattress system. AB - Pressure ulcers are distressing, frequently occur in immobile and malnourished patients and the cost to patients in terms of pain and anguish must be a prime consideration. There is little information on the incidence and prevalence of pressure ulcer development within the independent sector, as this is difficult to obtain. However, with advances in treatment and technology, combined with more people with complex and multiple needs being cared for at home, the requirement for specialist pressure-relieving support surfaces in the community is increasing. This article reviews the need for clinical and cost-effective pressure-relieving surface solutions in the community and introduces the concept of the Russka pressure-relieving low air-loss system. PMID- 15126978 TI - Lysosomal storage disorders: the need for better pediatric recognition and comprehensive care. PMID- 15126979 TI - Gaucher disease: lessons from a decade of therapy. PMID- 15126980 TI - Fabry disease in childhood. PMID- 15126981 TI - The mucopolysaccharidoses: a heterogeneous group of disorders with variable pediatric presentations. PMID- 15126982 TI - Pompe disease in infants and children. PMID- 15126983 TI - Achieving closure through disclosure: experience in a pediatric institution. PMID- 15126984 TI - Treatment of respiratory system (not just lung!) abnormalities in Mucopolysaccharidosis I. PMID- 15126985 TI - Adding to the health services research toolbox. PMID- 15126986 TI - Primary prevention of atopic dermatitis in breast-fed infants: what is the evidence? PMID- 15126987 TI - A tale of twins and insulin resistance. PMID- 15126988 TI - Effective treatment of alpha-mannosidosis by allogeneic hematopoietic stem cell transplantation. AB - OBJECTIVES: To study the efficacy of hematopoietic stem cell transplantation (HCT) for ameliorating the clinical manifestations of alpha-mannosidosis. STUDY DESIGN: Four patients with alpha-mannosidosis underwent allogeneic HCT at the University of Minnesota. Diagnosis was established by assay of leukocyte alpha mannosidase activity level. Physical features, donor engraftment, leukocyte alpha mannosidase activity, neuropsychologic function, and hearing were monitored before and after transplantation, with follow-up ranging from 1 to 6 years. RESULTS: All 4 patients showed slowing of their neurocognitive development and sensorineural hearing loss before HCT. All patients are alive, with normalization of leukocyte enzyme activity after HCT. Intellectual function has stabilized, with improvement in adaptive skills and verbal memory function in 3 of 4 patients. Hearing has improved to normal or near normal for speech frequencies in 3 patients. No new skeletal abnormalities have developed. CONCLUSIONS: HCT can halt the progressive cognitive loss in patients with alpha-mannosidosis. Early diagnosis and treatment with HCT is critical for optimal results. PMID- 15126989 TI - Enzyme replacement therapy in mucopolysaccharidosis VI (Maroteaux-Lamy syndrome). AB - OBJECTIVES: To evaluate the safety and efficacy of weekly treatment with human recombinant N-acetylgalactosamine 4-sulfatase (rhASB) in humans with mucopolysaccharidosis type VI (MPS VI). STUDY DESIGN: An ongoing Phase I/II, randomized, two-dose, double-blind study. Patients were randomized to weekly infusions of either high (1.0 mg/kg) or low (0.2 mg/kg) doses of rhASB. Six patients (3 male, 3 female; age 7-16 years) completed at least 24 weeks of treatment, five of this group have completed at least 48 weeks. RESULTS: No drug related serious adverse events, significant laboratory abnormalities, or allergic reactions were observed in the study. The high-dose group experienced a more rapid and larger relative reduction in urinary glycosaminoglycan that was sustained through week 48. Improvements in the 6-minute walk test were observed in all patients with dramatic gains in those walking <100 meters at baseline. Shoulder range of motion improved in all patients at week 48 and joint pain improved in patients with significant pain at baseline. CONCLUSIONS: rhASB treatment was well-tolerated and reduced lysosomal storage as evidenced by a dose dependent reduction in urinary glycosaminoglycan. Clinical responses were present in all patients, but the largest gains occurred in patients with advanced disease receiving high-dose rhASB. PMID- 15126990 TI - Enzyme replacement therapy for mucopolysaccharidosis I: a randomized, double blinded, placebo-controlled, multinational study of recombinant human alpha-L iduronidase (laronidase). AB - OBJECTIVE: To confirm the efficacy and safety of recombinant human alpha-L iduronidase (laronidase) in patients with mucopolysaccharidosis I (MPS I). STUDY DESIGN: This was a randomized, double-blinded, multinational study of 45 patients with MPS I administered 100 U/kg (0.58 mg/kg) laronidase, or placebo intravenously weekly for 26 weeks. The coprimary efficacy end points compared the median change from baseline to week 26 between groups in percentage of predicted normal forced vital capacity (FVC) and in 6-minute walk test (6MWT) distance through the use of the Wilcoxon rank sum test. RESULTS: The laronidase (n=22) and placebo (n=23) groups had similar baseline characteristics. After 26 weeks, patients receiving laronidase compared with placebo showed mean improvements of 5.6 percentage points in percent of predicted normal FVC (median, 3.0; P=.009) and 38.1 meters in 6MWT distance (median, 38.5; P=.066; P=.039, analysis of covariance). Laronidase also significantly reduced hepatomegaly and urinary glycosaminoglycans, and, in more severely affected patients, improved sleep apnea/hypopnea and shoulder flexion. Laronidase was well-tolerated. Nearly all patients receiving enzyme had development of IgG antibodies, without apparent clinical effects. CONCLUSIONS: In patients with MPS I, laronidase significantly improves respiratory function and physical capacity, reduces glycosaminoglycan storage, and has a favorable safety profile. PMID- 15126991 TI - National estimates of hospital utilization by children with gastrointestinal disorders: analysis of the 1997 kids' inpatient database. AB - OBJECTIVES: To identify and to generate national estimates of the principal gastrointestinal (GI) diagnoses associated with hospital utilization and to describe national hospital utilization patterns associated with pediatric GI disorders. STUDY DESIGN: We analyzed a nationwide and stratified probability sample of 1.9 million hospital discharges from 1997 of children 18 years and younger, weighted to 6.7 million discharges nationally. Principal GI diagnoses were identified through the use of the Clinical Classification Software and Major Diagnostic Categories. RESULTS: In 1997 in the United States, there were 329,825 pediatric discharges associated with a principal GI diagnosis, accounting for more than 2.6 billion US dollars in hospital charges and more than 1.1 million hospital days. Appendicitis, intestinal infection, noninfectious gastroenteritis, abdominal pain, esophageal disorders, and digestive congenital anomalies combined accounted for 75.1% of GI discharge diagnoses, 64.2% of GI hospital charges, and 68.0% of GI hospital days. Excluding normal newborn infants and conditions related to pregnancy, GI disorders were the third leading cause of hospitalization. CONCLUSIONS: GI disorders are a leading cause of hospitalization of children. A minority of GI conditions account for the majority of measures of utilization. Children are hospitalized for GI conditions and at institutions that are distinct from adults. PMID- 15126992 TI - Importance of organ dysfunction in determining hospital outcomes in children. AB - OBJECTIVES: To use measures of organ dysfunction derived from administrative data to assess clinical and economic outcomes in hospitalized children. STUDY DESIGN: We used the International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) diagnostic and procedure codes to evaluate organ dysfunction in all patients, excluding neonates, in the Healthcare Cost and Utilization Project Kids' Inpatient Database (KID). We adapted consensus clinical definitions to characterize organ dysfunction in terms of degree of impairment, type of organ system involvement, and number of dysfunctional organ systems. Univariate and multivariable models were constructed to determine the impact of organ dysfunction on in-hospital mortality and resource use. RESULTS: Patients with organ dysfunction (n=51,386) were younger and more often male than those without organ dysfunction, and they had significantly higher in-hospital mortality and resource use. Organ dysfunction, assessed in terms of degree, type, and number of dysfunctional organ systems, was consistently associated with all hospital outcomes. In multivariable models, types of organ system failures were most predictive of in-hospital mortality, whereas degree of organ system involvement allowed for a better assessment of resource use. CONCLUSIONS: Administrative data can be used to characterize multiple dimensions of organ dysfunction in children. Hospitalizations involving organ dysfunction are associated with significant clinical and economic consequences. PMID- 15126993 TI - Effect of breast-feeding on the development of atopic dermatitis during the first 3 years of life--results from the GINI-birth cohort study. AB - OBJECTIVE: To investigate if exclusive breast-feeding for 4 months is associated with atopic dermatitis during the first 3 years of life. STUDY DESIGN: Data on 3903 children were taken from yearly parental-administered questionnaires from a birth cohort study in Germany (recruited 1995-1998) comprised of a noninterventional (NI) and an interventional (I) subgroup. Outcomes were physician-diagnosed atopic dermatitis (AD) and itchy rash. Multiple logistic regression was performed for the entire cohort and stratified by family history of allergy and by study group adjusting for a fixed set of risk factors for allergies. RESULTS: Exclusive breast-feeding (52 % of children) was not associated with higher risk for AD either in the entire cohort (OR(adj,) 0.95; 95% CI, 0.79-1.14) or if stratified by family history of AD. In the I subgroup, but not in the NI subgroup, exclusive breast-feeding showed a significant protective effect on AD if compared with conventional cow's milk formula (OR(adj), 0.64; 95% CI, 0.45-0.90). CONCLUSION: These findings do not support the hypothesis that exclusive breast-feeding is a risk factor for development of atopic dermatitis but is protective if compared with conventional cow's milk. Observational studies might not be able to effectively control for selection bias and reverse causation. PMID- 15126994 TI - Insulin resistance in healthy prepubertal twins. AB - OBJECTIVES: To evaluate insulin sensitivity (S(I)) in prepubertal twins and to examine the relation to reduced birth weight, prematurity, and peroxisome proliferator-activated receptor-gamma (PPAR gamma) polymorphism. STUDY DESIGN: Fifty twins (birth weight SDS, -0.7 +/- 0.2; gestation, 33.5 +/- 0.5 weeks; and body mass index SDS, -0.04 +/- 0.2) were studied at 8.2 +/- 0.3 years. S(I) was measured by Bergman's minimal model from a 90 minutes frequently sampled intravenous glucose test. Twenty control children (height SDS, -1.7 +/- 0.3; birth weight SDS, -0.3 +/- 0.2; and gestation of 39.2 +/- 0.7 weeks) were also evaluated at 7.0 +/- 0.4 years. The PPAR gamma T-variant polymorphism was evaluated in 41 twins. Values are expressed as mean +/- SEM, or 95% confidence intervals. RESULTS: S(I) was reduced in twins compared with control subjects, (12.7 [11-15] versus 23.0 [16.8-31.4] 10(-4) min(-1) microU/mL, respectively, P=.005). The reduction in S(I) was independent of prematurity and birth weight and zygosity (P<.0001). There was no difference in S(I), even in twin pairs with >20% difference in birth weight (P=.9). The PPAR gamma heterozygote T-variant polymorphism was present in 7 of 41, with a further reduction in S(I) (P=.03) and a later gestation (P=.03). These twins also had increased fat mass (P=.02) but with similar fat free mass (P=.14). CONCLUSIONS: Twin children, independent of prematurity or birth weight, had a marked reduction in S(I). To use twins as a model to study the fetal origins of adult diseases for glucose homeostasis is not valid. PMID- 15126995 TI - Reduced bone mineral density and increased bone turnover in Prader-Willi syndrome compared with controls matched for sex and body mass index--a cross-sectional study. AB - OBJECTIVES: To study bone mineral status, body composition, and biochemical markers of bone turnover in Prader-Willi syndrome (PWS). STUDY DESIGN: Eight subjects with PWS (three males, five females; mean age, 24 years [range 16-41]) were included. Each subject was compared with an age-, sex- and body mass index matched control randomly drawn from the background population. Bone mineral density (BMD), lean body mass, and fat mass were measured. Plasma PINP, PIIINP, osteocalcin, total alkaline phosphatase, bone-specific alkaline phosphatase, C terminal telopeptide of type I collagen, and urine cross-linked N-terminal telopeptide of type I collagen were measured as biochemical markers of bone and collagen turnover. RESULTS: The PWS patients had significantly lower whole-body BMD (mean +/- SD, 1.020 +/- 0.041 vs 1.237 +/- 0.118 g/cm(2); 2p <.01) than controls due to lower bone mineral content (BMC: 2291 +/- 607 vs 2825 +/- 409 g; 2p=.02). Resorptive and formative bone markers were significantly elevated in patients compared with controls. Plasma testosterone was low in male patients (3.50 +/- 4.97 vs 19.2 +/- 8.78 nmol/L, 2p=.05), whereas no difference in plasma estradiol was present. CONCLUSIONS: The patients had a low BMD due to a high bone turnover. This high turnover was probably linked to sex steroid deficiency. PMID- 15126996 TI - Effects of exercise training on vascular function in obese children. AB - OBJECTIVES: Atherosclerosis is a disease that begins in childhood; endothelial dysfunction is its earliest detectable manifestation, and primary prevention strategies are likely to be most effective if instituted early. The aim of this study was to characterize the impact of obesity on vascular function in young children and to determine whether an exercise program improves abnormalities in vascular function. STUDY DESIGN: The influence of 8 weeks of exercise training was examined in 14 obese subjects, 8.9 +/- 0.4 years of age, with the use of a randomized crossover protocol. Conduit vessel endothelial function was assessed by means of high-resolution ultrasound and flow-mediated dilation of the brachial artery (FMD). RESULTS: Exercise training did not change subcutaneous fat mass, body weight, or body mass index. FMD in the obese group was significantly impaired relative to matched control subjects at entry (6.00% +/- 0.69% to 12.32% +/- 3.14%, P <.0001). FMD significantly improved with exercise training (7.35% +/ 0.99%, P <.05) in the obese group. CONCLUSIONS: Conduit vessel FMD, a validated surrogate measure of early atherosclerosis, was impaired in obese children but improved as a result of exercise training. This study supports the value of an exercise program in the treatment of obese children in a primary prevention setting. PMID- 15126997 TI - Adverse events associated with neonatal exchange transfusion in the 1990s. AB - OBJECTIVE: To determine the rates of adverse events associated with neonatal exchange transfusions performed for hyperbilirubinemia. STUDY DESIGN: Retrospective chart review of 55 neonates who underwent 66 exchange transfusions at two perinatal centers in Cleveland between 1992 and 2002. Demographic data, causes of jaundice, details of exchange method, and adverse events occurring within one week of exchange were recorded. At the time of exchange, 62% of infants had other neonatal morbidities. Outcomes were stratified according to gestational ages < or =32 weeks, 33 to 36 weeks, and > or =37 weeks. RESULTS: Overall, 74% of exchanges were associated with an adverse event. The most common events were thrombocytopenia (44%), hypocalcemia (29%), and metabolic acidosis (24%), of which 69%, 74%, and 44%, respectively, required treatment. There were two serious adverse events, both in infants with other serious neonatal morbidities: seizures in one infant and the death of a critically ill preterm infant (body weight 731 g, gestational age 25 weeks). There were no cases of sepsis, necrotizing enterocolitis, or cardiac arrest. Adverse events were more frequent in exchanges done on preterm infants: < or =32 weeks (87%), 33 to 36 weeks (78%), and > or =37 weeks (67%), and in infants with other neonatal morbidity (79% vs 57%; P=.08). Controlling for neonatal morbidity, we found the odds of an adverse event were significantly higher when both umbilical venous and arterial catheters were used compared with other methods of exchange (88% vs 58%; OR, 5.17; 95% CI, 1.1, 34.2; P=.03). CONCLUSIONS: The majority of adverse events associated with exchange transfusion are laboratory abnormalities and are asymptomatic and treatable. PMID- 15126998 TI - Tissue transglutaminase autoantibody detection in human saliva: a powerful method for celiac disease screening. AB - OBJECTIVE: To test the possibility of detecting tissue transglutaminase autoantibodies (tTG-Abs) in saliva with a novel sensitive fluid-phase radioimmunoassay (RIA). STUDY DESIGN: Paired saliva and serum samples from 39 patients with celiac disease (CD), at the first biopsy (Group 1: 28 females, mean age 11.5 +/- 11.1 years); 32 controls with a normal duodenal mucosa (Group 2: 18 females, mean age 8.1 +/- 3.6 years); and 32 healthy volunteers (Group 3: 21 females, mean age 31.7 +/- 9.8 years) were studied for tTG-Ab presence. Limit of positivity for salivary assay was calculated according to the 99th percentiles of Group 2 control children and was expressed as an autoantibody (Ab) index. RESULTS: Salivary tTG-Abs were found in 97.4% of the patients with CD and in 100% of the corresponding serum samples. All Group 3 subjects were negative with both saliva and serum assays. A correlation between saliva and serum tTG-Ab titers was found (r=0.826, P=.0014). CONCLUSIONS: This study demonstrates that it is possible to detect salivary tTG-Abs in CD with a non-invasive, simple to perform, reproducible and sensitive method. PMID- 15127000 TI - Brain damage in glycogen storage disease type I. AB - OBJECTIVE: To investigate brain morphology and function in patients with glycogen storage disease type I (GSDI). STUDY DESIGN: Nineteen patients (13 females and 6 males, aged 0.9-22.6 years) and 38 sex- and age-matched controls entered the study. Neurological examinations, psychometric tests (IQ, tests of performance and verbal abilities), standard electroencephalogram (EEG), somatosensory (SEPs), visual (VEPs), and brain-stem auditory evoked potentials (BAEPs), and brain magnetic resonance imaging (MRI) were performed. RESULTS: The results of tests of performance ability were lower in patients than in controls (P <.05). The prevalence of abnormal EEG findings (26.3% versus 2.6%), VEPs (38.4% versus 7.7%), SEPs (23.0% versus 0%), and BAEPs abnormalities (15.7% versus 0%) was higher in patients than in controls (P <.05). MRI pattern was altered in 57.1% of patients and was normal in all controls (P <.05). Both results of tests of performance ability and BAEPs abnormalities significantly correlated with the frequency of admissions for hypoglycemia, whereas EEG abnormalities correlated with dietary compliance (P <.05). CONCLUSIONS: Brain damage, probably caused by recurrent severe hypoglycemia, may be present in patients with GSDI. PMID- 15127002 TI - Definitive diagnosis in children with congenital hypothyroidism. AB - OBJECTIVES: To investigate the definitive diagnosis and underlying causes of congenital hypothyroidism (CH) in eligible children through the use of a standardized protocol. STUDY DESIGN: Children > or =3 years of age with CH without an identified permanent cause underwent a diagnostic algorithm. Eligible subjects had an anatomically normal thyroid or had not undergone imaging studies. After thyroxine was discontinued for 4 weeks, thyroid function tests and a thyroid ultrasound were obtained. An abnormal ultrasound was followed by a (99m)Tc thyroid scan. A perchlorate washout test was performed in subjects with a normal ultrasound but abnormal thyroid function tests. Children with normal results were followed for 1 year. RESULTS: Of 33 children, 17 were boys. Nine (27%) had an absent or ectopic thyroid, 12 (36%) had dyshormonogenesis, and 12 (36%) had transient CH. Average thyroxine dose before medication discontinuation was 2.9 +/- 0.83 microg/kg in permanent cases versus 2.0 +/- 0.53 microg/kg in transient (P <.002). No complications from discontinuation of thyroxine occurred. CONCLUSIONS: A significant percentage of children with CH have a transient requirement for thyroid hormone. A standardized protocol with thyroid ultrasonography is a safe and sensitive approach to a trial off of thyroxine in select patients. PMID- 15127004 TI - hGH for short stature: ethical issues raised by expanded access. PMID- 15127005 TI - Nucleated red blood cells in preterm infants who have necrotizing enterocolitis. AB - Absolute nucleated red blood cell counts for 23 preterm newborn infants who subsequently had development of necrotizing enterocolitis were significantly higher than for 23 control infants. These infants may have been exposed to relative intrauterine hypoxemia, a possible risk factor for the development of necrotizing enterocolitis. PMID- 15127006 TI - Progressive beta cell failure in type 2 diabetes mellitus of youth. AB - The pathophysiology of type 2 diabetes mellitus (T2DM) involves insulin resistance and insulin deficiency. We report the progression in insulin sensitivity and secretion over a 6-year period in an adolescent with T2DM. This report further demonstrates that the earliest abnormality in youth T2DM is insulin resistance followed by progressive beta-cell failure. PMID- 15127007 TI - Predictors of control of diabetes: monitoring may be the key. AB - The factors affecting glycated hemoglobin concentration were examined in a cohort of children attending diabetes camp. Multivariate regression analysis revealed that frequency of self-monitoring of blood glucose correlated with lower glycated hemoglobin, whereas the number of insulin types used correlated with increased glycated hemoglobin. Frequent self-monitoring of blood glucose promotes long-term metabolic control. PMID- 15127008 TI - Steatohepatitis and unsuspected micronodular cirrhosis in Dorfman-Chanarin syndrome with documented ABHD5 mutation. AB - Mutation in ABHD5 causes Dorfman-Chanarin syndrome (DCS), a multisystem triglyceride storage disorder. Ultrastructural study of leukocytes confirmed DCS in a child homozygous for a novel ABHD5 mutation, with ichthyosis, developmental delay, and steatohepatitis with cirrhosis, manifest only as elevated aminotranferase levels. We recommend early assessment for liver disease in DCS. PMID- 15127010 TI - Neurofibromatosis in children with Rhabdomyosarcoma: a report from the Intergroup Rhabdomyosarcoma study IV. AB - We identified all children enrolled in the Intergroup Rhabdomyosarcoma Study Group-IV with neurofibromatosis type I (NF1) and rhabdomyosarcoma. Among 1025 eligible patients, 5 (0.5 %) had NF1. Three children had relapses, two of whom died of progressive disease. Patients with NF1 and rhabdomyosarcoma should be treated with intensive contemporary therapy protocols. PMID- 15127011 TI - Toxic ingestion of 6-mercaptopurine by young siblings of pediatric oncology patients. AB - We report two cases of accidental ingestion of large quantities of 6 mercaptopurine (6-MP) in siblings of patients with acute lymphoblastic leukemia (ALL). These cases reinforce the need for thorough anticipatory guidance to families in order to prevent the incidence of potentially life-threatening accidental ingestions. PMID- 15127012 TI - Acquired Glanzmann's thrombasthenia as part of multiple-autoantibody syndrome in a pediatric heart transplant patient. AB - Although advances in immunosuppressive therapy have allowed prolonged patient survival, immune dysregulation observed in these patients has increased. We report an 11-year-old female heart transplant recipient in whom Glanzmann thrombasthenia was identified as part of a "multiple autoantibody syndrome" manifesting sequentially as autoimmune hemolytic anemia, thrombocytopenia, and neutropenia. PMID- 15127013 TI - Stuttering priapism associated with withdrawal from sustained-release methylphenidate. AB - Stuttering priapism is intermittent, prolonged, painful, pathologic erections with intervening periods of detumescence. An adolescent had stuttering priapism associated with withdrawal from sustained-release methylphenidate. To our knowledge, this is the first such report of stuttering priapism associated with stimulant drugs for treatment of attention deficit hyperactivity disorder. PMID- 15127014 TI - Severe vesicoureteral reflux and chronic renal failure: a condition peculiar to male gender? Data from the ItalKid Project. AB - Primary vesicoureteral reflux (VUR), one of the principal causes of chronic renal failure (CRF), occurs as a result of two distinct and sex-related mechanisms: congenital renal hypoplasia, which is prevalent in males, and acquired renal scarring in females. We used data from the ItalKid Project, a prospective population-based CRF registry of patients undergoing conservative treatment, to evaluate the gender distribution and severity of primary VUR, the age at diagnosis, and the diagnostic and therapeutic methods adopted in children with CRF. The prevalence of males (77.5%), the severity of VUR (grade IV-V), and the early age at diagnosis (18% prenatally) seem to suggest that congenital renal damage is the major cause of pediatric CRF. PMID- 15127015 TI - Anomalous systemic venous return. PMID- 15127016 TI - Nonsurgical removal of a knotted pediatric femoral vein catheter. PMID- 15127021 TI - Controlled slicing in the management of congenitally missing second premolars. AB - This report describes a simple method of allowing permanent first molars to drift mesially in patients with congenitally missing second premolars, thus facilitating future orthodontic treatment. Controlled slicing of the deciduous second molar between the ages of 8 and 9 years produced a bodily controlled mesial movement of the permanent first molar in less than 1 year with no or minor rotations or inclination. These results are compared with controlled slicing in 10- to 11-year-olds and with treatment involving extraction alone. PMID- 15127020 TI - A comparison of current prediction imaging programs. AB - To investigate perceived differences in the ability of current software to simulate the actual outcome of orthognathic surgery, we chose 10 difficult test cases with vertical discrepancies and "retreated" them using the actual surgical changes. Five programs--Dentofacial Planner Plus, Dolphin Imaging, Orthoplan, Quick Ceph Image, and Vistadent--were evaluated, by using both the default result and a refined result created with each program's enhancement tools. Three panels (orthodontists, oral-maxillofacial surgeons, and laypersons) judged the default images and the retouched simulations by ranking the simulations in side-by-side comparisons and by rating each simulation relative to the actual outcome on a 6 point scale. For the default and retouched images, Dentofacial Planner Plus was judged the best default simulation 79% and 59% of the time, respectively, and its default images received the best (lowest) mean score (2.46) on the 6-point scale. It also scored best (2.26) when the retouched images were compared, but the scores for Dolphin Imaging (2.83) and Quick Ceph (3.03) improved. Retouching had little impact on the scores for the other programs. Although the results show differences in simulation ability, selecting a software package depends on many factors. Performance and ease of use, cost, compatibility, and other features such as image and practice management tools are all important considerations. Users concerned with operating system compatibility and practice management integration might want to consider Dolphin Imaging and Quick Ceph, the programs comprising the second tier. PMID- 15127022 TI - Clinical response to allergies in patients. AB - Allergenic reactions are becoming a more prevalent concern of practitioners in health-related fields. As patient susceptibility increases, the need for understanding and successfully treating such heterogeneous responses becomes paramount. This article presents a theoretical bioengineering model that considers the influences of specific concentrations on their chemical effects. This model then is applied to contemporary orthodontics in which nickel and, to a lesser extent, chromium are antagonists in as many as 1 in 500 patients-which, for North America, amounts to about 4000 patients per year. For the occasional patient who responds positively to a questionnaire, a skin patch test would be administered to establish whether alternative alloys could be used, whether they be gold, platinum, nickel-free stainless steel, or titatnium-molybdenum alloys. PMID- 15127023 TI - Effects of an anteriorly titrated mandibular position on awake airway and obstructive sleep apnea severity. AB - The objective of this study was to investigate whether a reduction of obstructive sleep apnea (OSA) severity was associated with significant upper airway (UA) changes after an anterior titration of the mandibular position. Eighteen OSA patients with a mean (SD) apnea hypopnea index (AHI) of 32.5 (12.3) were recruited. Baseline supine cephalometry was obtained before the initial insertion, and follow-up supine cephalometry was undertaken after titration with a titratable oral appliance in place. The mean AHI before treatment was significantly reduced to 9.7 (7.4) (P <.001) after titration. In 13 responders with AHI reduced to < or =15/h, a significant forward displacement of the anterior wall of the velopharynx (P <.05) was observed. In addition, there was a significant forward displacement of the posterior wall of the oropharynx and the hypopharynx (P <.05). In the 5 nonresponders, no significant changes in the position of the anterior and posterior wall were observed. There was no significant difference in the total amount of mandibular advancement between responders and nonresponders. We conclude that treatment success with oral appliance therapy appears to depend not only on anterior titration of the mandibular position to enlarge the UA, but also on the amount of change in the size of the UA in response to mandibular advancement. PMID- 15127024 TI - Long-term changes in pharyngeal airway morphology after mandibular setback surgery. AB - Long-term changes in pharyngeal airway morphology were evaluated after mandibular setback surgery in 10 women who were diagnosed as having skeletal mandibular prognathism and underwent mandibular setback surgery by bilateral sagittal split ramus osteotomy (SSRO) and orthodontic multi-bracket treatment. The subjects were assessed before treatment (T1), 3-6 months after SSRO (T2), and 2 or more years after SSRO (T3). From T1 to T2, the pharyngeal airway constricted significantly. On the other hand, from T2 to T3, the lower facial morphology showed no significant changes. The pharyngeal airway morphology showed significant changes in soft-palate length and posterior reference line to point of posterior tongue, indicating a tendency for relapse. These results suggest that, although the lower facial morphology and the pharyngeal airway morphology showed marked changes after SSRO, the pharyngeal airway morphology exhibited gradual physiologic readaptation. However, because the lower facial morphology after surgery was retained and stable for a long period, it was considered normal for the pharyngeal airway morphology to adapt after surgery to the improved hard tissue relationship. PMID- 15127025 TI - Comparison of tongue functions between mature and tongue-thrust swallowing--an ultrasound investigation. AB - Many studies have proved that tongue thrusting plays a significant role in the etiology of some orofacial deformities. To learn more about the relationship between tongue function and the form of orofacial structures, it is important to recognize patients with abnormal swallowing patterns. A cushion scanning technique in combination with M-mode ultrasound was applied to measure and compare tongue movements between mature swallowing and tongue-thrust swallowing. Forty subjects with a mature swallowing pattern (23 female, 17 male, aged 21.8 +/ 6.1 years) and 15 subjects with a tongue-thrust swallowing pattern (8 female, 7 male, aged 13.6 +/- 6.6 years) were included in this study. Duration, range, speed, and reproducibility for each of the swallowing subphases were calculated and compared. Tongue-thrust swallowers had a longer late transport phase than mature swallowers (P <.0009), and the tongue speed was faster in the early final phase (P <.05) compared with mature swallowers. Distinctly different movements can be positively differentiated with the method used. PMID- 15127026 TI - Morphometrics for cephalometric diagnosis. AB - This article demonstrates morphometric methods by applying them to an orthodontic sample. A total of 150 pretreatment cephalograms of consecutive patients (84 female, 66 male) were traced and digitized. Fifteen points were used for the analysis. The tracings were superimposed by the Procrustes method, and shape variability was assessed by principal component analysis. Approximately 70% of the total sample variability was incorporated in the first 5 principal components. The most significant principal component, accounting for 29% of shape variability, was the divergence of skeletal pattern; the second principal component, accounting for 20% of shape variability, was the anteroposterior maxillary relationship. It is recommended that Procrustes superimposition and principal component analysis be incorporated into routine cephalometric analysis for more valid and comprehensive shape assessment. PMID- 15127027 TI - Objective assessment of occlusal and coronal characteristics of untreated normals: a measurement study. AB - To determine whether tooth shape and position are constant within tooth types, dental casts of 68 Indians (age range, 10-32 years; mean, 18.72 years) were analyzed. The casts were selected from a larger sample and met the following criteria: Class I molar and canine relationships; overjet and overbite within normal limits; well-related vertical, transverse, and anteroposterior relationships with pleasing profiles and well-aligned arches; and no supernumerary teeth or large restorations. None of the subjects had received orthodontic treatment, and all were in good health and exhibited normal growth. Crown angulation, inclination, offset of maxillary molar, curve of Spee, crown facial prominence, horizontal crown contour, and vertical crown contour were assessed. Means, standard deviations, and standard errors were calculated. The measurements were compared with Andrews's data on 120 nonorthodontic normal occlusion casts. The Student t test was used to determine the significance of differences between the 2 sets of data. It was found that teeth of the same tooth type have similar values of horizontal and vertical crown contours. Values for inclination, angulation, and relative prominence were also similar. Hence, we concluded that tooth shape and position are constant for each tooth type. The data from this study were comparable with Andrews's findings. However, from this study, it would be safe to presume that all teeth except the maxillary second molars require alterations in the bracket base inclination value, and that the maxillary lateral incisor, canine, second premolar, and second molar, and the mandibular canine, require alterations in angulation values. PMID- 15127028 TI - Changes in the curve of Spee with treatment and at 2 years posttreatment. AB - The purposes of this study were to determine pretreatment skeletal and dental patterns related to curve of Spee depth, evaluate changes in the curve of Spee with treatment and the effects on dentofacial structures, and determine factors associated with the stability of the curve of Spee after treatment. Lateral cephalograms and dental casts were evaluated for 24 white male and 26 white female patients at pretreatment, posttreatment, and at least 2 years postretention. All patients had mandibular permanent second molars and premolars erupted and in occlusion. The following variables were correlated with increased pretreatment curve of Spee depth: low Frankfort-mandibular plane angle, deep overbite, increased overjet, and Class II molar malocclusion. No significant differences were found in pretreatment curve depth between male and female patients or between right and left sides. Leveling the curve of Spee was accomplished by uprighting the molars, extruding the premolars, and intruding or flaring the incisors. Extraction and nonextraction cases demonstrated no significant differences in the amount of curve relapse. Increased curve relapse was correlated with postretention increases in overbite, irregularity index, and patients given removable retainers. Overall, the curve of Spee was relatively stable after treatment, with only a 16% relapse of the leveled curve. PMID- 15127029 TI - Association between vertical development of the cervical spine and the face in subjects with varying vertical facial patterns. AB - The upper cervical spine and face were compared longitudinally from 6 to 15 years of age in 2 groups of children with low (n = 16) and high (n = 14) MP-SN angles. The purpose of the study was to assess morphological and incremental associations between the vertical development of the cervical spine and the face in subjects with varying vertical facial patterns. Morphological associations were not detected during the observation period. Long faces were found on children with short necks, and short, square faces on those with long necks. Vertical growth of the upper cervical spine and face were weakly correlated in the 6-to-12-year period but strongly correlated later during puberty. The vertical distance between gonion and the body of the second cervical vertebra was remarkable for its constancy during childhood and puberty, indicating that gonion and the second cervical vertebra body were anatomically interrelated. This anatomical relationship strongly suggests that a mutual relationship also exists between vertical growth of the upper cervical spine and the face, especially the lower face. In addition, the lower face had the ability for independent vertical growth that was not coordinated with vertical cervical growth. This was the case in the 6-to-12-year period, but not later. PMID- 15127030 TI - Comparison of the effects of 1,25 dihydroxycholecalciferol and prostaglandin E2 on orthodontic tooth movement. AB - This study compared the effects of local administrations of prostaglandin E2 (PGE2) and 1,25-dihydroxycholecalciferol (1,25-DHCC) on orthodontic tooth movement in rats. Thirty-seven 6-week-old male Sprague-Dawley rats, weighing 160 +/- 10 g were used. Five rats served as the baseline control group. A fixed appliance system exerting 20 g of distally directed force was applied on the maxillary incisors of 32 animals for 9 days. Eight rats served as the appliance control group; 8 received a 20-microL injection of dimethyl sulfoxide (solvent for 1,25-DHCC) on days 0, 3, and 6; 8 received 20 microL of 10(-10) mol/L 1,25 DHCC on days 0, 3, and 6; 8 received a single injection of 0.1 mL of 0.1 microg PGE2 only on day 0. There was no significant difference in tooth movement between the PGE2 and the 1,25-DHCC groups. Both PGE2 and 1,25-DHCC enhanced the amount of tooth movement significantly when compared with the control group. The numbers of Howship's lacunae and capillaries on the pressure side were significantly greater in the PGE2 group than in the 1,25-DHCC group. On the other hand, the number of osteoblasts on the external surface of the alveolar bone on the pressure side was significantly greater in the 1,25-DHCC group than in the PGE2 group. Thus, 1,25 DHCC was found to be more effective in modulating bone turnover during orthodontic tooth movement, because its effects on bone formation and bone resorption were well balanced. PMID- 15127031 TI - Focal hyalinization during experimental tooth movement in beagle dogs. AB - The aim was to study morphological differences between the periodontal structures of beagle dogs showing different rates of tooth movement under identical experimental conditions. An orthodontic appliance was placed on the mandibular second premolar and the first molar to exert a continuous and constant reciprocal force of 25 cN. Tooth movement was recorded weekly. The dogs were killed after 1, 4, 20, 40, and 80 days for histological evaluation. Haematoxylin and eosin staining was used for tissue survey, alkaline phosphatase staining was used as a marker for active osteoblasts, and tartrate resistant acid phosphatase staining was used for osteoclasts. After 24 hours, osteoclastic and osteoblastic activity had already increased at the pressure and tension sides, respectively, and, in some samples, hyalinization was found. In case of fast-moving teeth, areas of direct bone resorption at the pressure side and deposition of trabecular bone at the tension side were found throughout the experimental period. In the periodontal ligaments of teeth showing little movement, small patches of hyalinization were found at the pressure side, mostly located buccally or lingually of the mesiodistal plane. These phenomena were found in both molars and premolars and at all time points. It is concluded that small focal hyalinizations might be a factor that could explain individual differences in the rate of tooth movement. PMID- 15127032 TI - Extraoral appliances: A twenty-first century update. AB - With the locations of the centers of resistance of the dentomaxillary complex having recently been defined, and with the newly acquired knowledge of the critical interplay between part-time extraoral and full-time intraoral force systems, the basic designs of all types of extraoral appliances are discussed in depth. Armed with this information, the clinician can improve the efficiency of all extraoral appliances to obtain better dentomaxillary-complex growth control and negate or enhance appliance-induced intraoral force systems. PMID- 15127033 TI - Practice efficiency: the customized treatment process. AB - In business, operating efficiently and achieving economies of scale are goals managers continuously strive to meet. In orthodontics, achieving these goals can mean treating more patients in less time and at a lower cost but with an outcome that meets the professional standard and meets or exceeds the patient's expectations. Efficiency allows the orthodontist to maximize the return on constrained capital assets. This article presents a treatment process designed to enhance efficiency. It outlines and discusses the processes of dynamic diagnosis, customized mechanics, patient integration in the treatment, and management of expectations. These elements combine to make an integrated system of treatment. Finally, this article will discuss how efficient treatment is also cost-effective treatment and contributes to continuous profit. PMID- 15127034 TI - Management of congenitally missing second premolars with orthodontics and single tooth implants. AB - This article describes the treatment of an adolescent girl who was congenitally missing all 4 second premolars and had a retained mandibular second primary molar. Various treatment alternatives are discussed, and the final treatment plan of space opening for 3 implants and space closure of the maxillary left second premolar site is presented. PMID- 15127035 TI - The OrthoCAD bracket placement solution. PMID- 15127036 TI - Litigation, legislation and ethics. Releases and minors. PMID- 15127040 TI - Rotor blade design. PMID- 15127041 TI - Pediatric supracondylar humerus fractures. PMID- 15127043 TI - Clinical features of patients intubated by a flight team. AB - INTRODUCTION: Little information exists about the clinical features of patients intubated by a flight program and the relationships of those factors with flight team intervention. METHODS: Prospective analysis was conducted of consecutive patients intubated by a helicopter flight team. RESULTS: The flight team intubated 11.1% of patients transported. Scene origin and trauma diagnosis were associated with flight team intubation (P <.01). The diagnosis distribution differed between scene and interfacility groups (P <.01), with a trauma diagnosis more common in scene patients and a medical diagnosis more common on interfacility flights. The most common clinical indicator for intubation on both scene and interfacility flights was mental status change. The distribution of indicators, however, also varied with patient origin (P <.01). CONCLUSION: The flight team most commonly intubated patients who originated at the scene, suffered traumatic injury, and had altered mental status. Both the diagnosis distribution and primary clinical indicator for intubation varied with patient origin. PMID- 15127042 TI - The 2003 Air Medical Leadership Congress: findings and recommendations. AB - To address important concerns facing the air medical community, 149 air medical transport leaders, providers, consultants, and experts met September 4-6, 2003, in Salt Lake City, Utah, for a 3-day summit-the Air Medical Leadership Congress: Setting the Health Care Agenda for the Air Medical Community. Using data from a Web-based survey, top air medical transport issues were identified in four core areas: safety, medical care, cost/benefit, and regulatory/compliance. This report reviews the findings of previous congresses and summarizes the discussions, findings, recommendations, and proposed industry actions to address these issues as set forth by the 2003 congress participants. PMID- 15127044 TI - Impact of a triage tool on air versus ground transport of cardiac patients to a tertiary center. AB - INTRODUCTION: The optimum method of transport for acute cardiac patients remains controversial. We proposed a physician-developed triage scheme for appropriate use of air versus mobile intensive care unit (ICU) in the transfer of cardiac patients and sought to determine the impact on the distribution of transport mode for cardiac patients in areas of personal characteristics and clinical factors and whether the triage scheme would be a valuable decision-making tool for physicians referring cardiac patients to tertiary centers. METHODS: This was a prospective, observational study of transport mode for cardiac patients transported to a tertiary care facility. A comparison was conducted with historical controls. The intervention studied was an educational program designed to teach a triage decision tool developed by a receiving cardiologist with input from the critical care transport team. Short-distance (less than 30 minutes) and long-distance transports were examined. A follow-up survey of referring hospitals was conducted. RESULTS: Short-distance transports enjoyed excellent compliance with 41 of 42 patients being transported by mobile ICU. Long-distance transports by mobile ICU increased from 55% to 65% during the study period. However, a third of the mobile ICU patients actually met air transport criteria. Long-distance patients transported by air had significantly higher transport costs, total hospital charges, and direct admission to the catheterization lab. Five of the 10 surveyed emergency department directors found the triage instrument useful in making transport decisions. CONCLUSIONS: A physician-developed triage instrument to select an appropriate mode of transport for acute cardiac transfers was effectively used. Further studies must validate the cardiac triage criteria against clinical outcomes, and more effective dissemination of the triage instrument must be sought. Furthermore, this information must be perceived as useful by referring physicians to gain wider acceptance. PMID- 15127052 TI - Creating a legacy of leadership. PMID- 15127053 TI - International networking--Internet style. PMID- 15127054 TI - Mapping the future of dietetics with geographic information systems. PMID- 15127055 TI - How to write for the public. PMID- 15127056 TI - Medicare reform: what it means to the future of dietetics. PMID- 15127057 TI - Barriers to the use of WIC services. AB - OBJECTIVES: To identify barriers that deter parents/caretakers of infants and children enrolled in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) from taking full advantage of the services provided by the program. SUBJECTS/SETTING: A total of 3,167 parents/caretakers at 51 New York State WIC local agency sites completed a barriers survey. DESIGN: Sixty eight potential barriers to WIC were identified through a literature review, five focus groups with parents/caretakers of WIC participants, and an expert review panel. The barriers survey was administered person-to-person to parents/caretakers of infants and children on WIC. Statistical analysis Classification tree analysis was used to identify characteristics that best predict WIC check usage behavior. RESULTS: A small set of barriers (n=11) were identified by more than 20% of respondents. Waiting too long was the most frequently cited barrier (48%). Difficulties in bringing the infant/child to recertify and rescheduling appointments were key variables associated with failure to use (ie, pick up or cash) WIC checks. Further analyses indicated that (a) for each additional reported barrier, there was a 2% increase in failure to use WIC checks (P<.0001); (b) waiting for services was related to an increase in the number of people who failed to use checks; and (c) the longer the reported wait, the greater the number of reported barriers (P<.0001). CONCLUSIONS: Conducting this barriers research enabled the New York State WIC to improve services provided to participants and their families. A decrease in waiting times should generally reduce exposure to noisy, crowded facilities and lead to fewer reports of nothing for kids to do. PMID- 15127058 TI - Improving WIC usage: office procedures, food package, and nutrition education. PMID- 15127059 TI - Validity of self-reported dietary intake at school meals by American Indian children: the Pathways Study. AB - OBJECTIVE: To examine the validity of a modified diet record-assisted 24-hour recall in third-grade (8 to 10 years old) American Indian children. DESIGN: The children were trained to record their food intake using diet records, and then they recalled their 24-hour food intakes, using the diet records as memory prompts, during interviews by trained staff using the Minnesota Nutrition Data System (NDS; version 2.6, 1993, Food database version 8A, Nutrient database version 23; Nutrition Coordinating Center, University of Minnesota, Minneapolis). The modified method added training in portion size estimation. Direct observation of the children's intakes during school meals was used to validate the accuracy of their self-reported recalls. SUBJECTS: Eighty third-grade children recruited from schools from four of the American Indian Nations participating in the Pathways Study. Statistical analyses performed Pearson correlations were used for nutrient level data. A mixed regression model (PROC MIXED), with no other fixed effects and site as a random effect, was used to test the null hypothesis that the difference between recalled and observed intakes was zero (H(o): beta(o)=0). Food intake data were obtained from the Nutrition Data System Record Reports. RESULTS: There were no significant differences between recalled and observed energy intakes for the school meals combined or for either meal individually. Percentages of energy intake from fat, protein, and carbohydrate from recalls were not significantly different from those observed for the combined school meals. Pearson correlations for energy and energy-providing nutrients ranged from 0.52 to 0.86 for both meals, from 0.55 to 0.86 for school lunch, and from 0.61 to 0.86 for school breakfast. Agreement between recalled and observed food items was 75%. Children recalled 57% of food quantities within +/-10% of observed quantities. CONCLUSIONS: At the group level, American Indian children were able to accurately report the macronutrient proportions of their total energy intake, and their reporting of total energy intake (+13% of criterion) compares favorably with that of other ethnic groups of children of similar age. They were able to accurately recall the majority of foods that they were independently observed consuming during school meals. PMID- 15127060 TI - Children's meal patterns have changed over a 21-year period: the Bogalusa Heart Study. AB - OBJECTIVE: The objective of this study was to analyze children's meal patterns over 2 decades. DESIGN: One 24-hour dietary recall was collected on each child who participated in one of seven cross-sectional surveys. SUBJECTS/SETTING: Dietary intake data were collected on 1,584 10-year-old children (65% white, 35% African American), in Bogalusa, LA, from 1973 to 1994. STATISTICAL ANALYSES: Descriptive statistics and one-way analysis of variance adjusting for gender and ethnicity. RESULTS: From 1973 to 1978, there was a marked increase (P<.0001) in the percentage of children who skipped breakfast, from 8.2% to 29.6%. When school breakfast was introduced in 1981, the proportion of children skipping breakfast declined to 12.5% (P<.01). From 1973-1974 to 1993-1994, the percentage of children eating a school lunch declined from 89.7% (1973-1974) to 78.2% (1993 1994) (P<.001); eating lunch brought from home increased from 5.9% to 11.1% (P<.01); consuming a home dinner decreased from 89.2% to 75.9% (P<.01); eating a dinner prepared outside the home increased from 5.4% to 19.0% (P<.01); consuming a meal at a restaurant increased from 0.3% to 5.4% (P<.0001); consuming snacks decreased (P<.0001); total eating episodes decreased from 6.6 to 5.2 (P<.0001); and eating time span significantly decreased from 12.4 hours to 11.5 hours (P<.0001). Despite these changes in meal patterns, no associations were found between meal patterns and overweight status. CONCLUSIONS: Striking alterations in the meal patterns of children occurred over the 2-decade period. These changes may have implications for the changes in the dietary intakes of children during the same time. However, data from this study do not support an association between meal patterns and children's overweight status. Further research with multiple days of assessment is needed to better understand the complexity of diet as it relates to childhood obesity. PMID- 15127061 TI - Development of a food frequency questionnaire to estimate calcium intake of Asian, Hispanic, and white youth. AB - OBJECTIVE: To develop a food frequency questionnaire (FFQ) that estimates calcium intake of Asian, Hispanic, and white youth living in the western United States. DESIGN: A list of 80 foods was assembled to create an FFQ to measure calcium intake. Evaluation of the FFQ spanned four consecutive weeks. An FFQ was completed during Week 1 and Week 4, and a 24-hour dietary recall was completed during Week 2 and Week 3.Subjects/setting A convenience sample of 162 Asian, Hispanic, and white youth ages 10 to 18 years was selected. Statistical analyses performed Percent agreement, paired t tests, Pearson correlation coefficients of cube-root transformed values, and deattenuated Pearson correlation coefficients of cube-root transformed values were used to evaluate the FFQ. RESULTS: The correlation between calcium intake estimates, when measured by first and second administrations of the FFQ, was 0.68 (Pearson's r) for the total sample. Correlations differed by age, sex, and ethnic subgroups as follows: 10 to 13 years (r=0.62), 14 to 18 years (r=0.73), male (r=0.73), female (r=0.64), Asian (r=0.77), Hispanic (r=0.72), and white (r=0.48). The correlation between calcium intakes as estimated by the second FFQ vs the average of the two 24-hour dietary recalls was 0.54 (deattenuated Pearson's r) for the total sample. This correlation differed by age, sex, and ethnic subgroups as follows: 10 to 13 years (r=0.46), 14 to 18 years (r=0.59), male (r=0.65), female (r=0.45), Asian (r=0.64), Hispanic (r=0.18), and white (r=0.57). CONCLUSIONS: A unique dietary survey has been developed to estimate calcium intake among Asian, Hispanic, and white youth in the United States. PMID- 15127062 TI - Developing dietary assessment tools. PMID- 15127063 TI - Dietary pattern change and acculturation of Chinese Americans in Pennsylvania. AB - OBJECTIVE: To obtain information about dietary pattern change of Chinese Americans in Pennsylvania and its relationship with demographic characteristics and acculturation indicators. DESIGN: A cross-sectional self-administered survey. SUBJECTS: A convenience sample of 399 Chinese Americans. Statistical analyses performed t Tests, analysis of variance with Tukey post-hoc tests, Spearman rank correlation, and chi(2) test. RESULTS: After immigration, Chinese Americans increased consumption frequency of all seven food groups (grains, vegetables, fruits, meat/meat alternatives, dairy products, fats/sweets, and beverages) and Western foods while consumption frequency of traditional Chinese foods decreased. Dietary variety also increased after immigration. Higher education and higher income levels were associated with a larger increase in consumption frequency of grains, vegetables, and fruits. Persons who resided in the United States for a longer period of time shared a greater increase in their consumption frequencies of vegetables, fats/sweets, and beverages. Persons with better English proficiency had a greater increase in their consumption frequency of grains, fruits, meat/meat alternatives, and fats/sweets. CONCLUSIONS: This study can help nutrition educators design appropriate educational programs for first-generation Chinese Americans that can facilitate the adoption of more healthful dietary practices. Nutrition educators should consider the dietary changes of Chinese American participants, such as skipping breakfast and increased consumption frequency of fats, sweets, and soft drinks, which were observed in this study. For example, acculturated first-generation Chinese Americans should be encouraged to decrease fats, sweets, and soft-drink consumption. Less-acculturated persons should be encouraged to maintain their healthful dietary pattern and increase consumption of vegetables and fruits. PMID- 15127064 TI - An evidence-based approach for dietitian prescription of multiple vitamins with minerals. AB - Dietitians working in hospitals are routinely involved in assessing energy and macronutrient (ie, protein, fat, carbohydrate) requirements of patients. However, complete nutritional therapy requires a comprehensive review of vitamin and mineral requirements. Scientific evidence for vitamin and mineral supplementation is primarily based on healthy, free-living people. This raises clinical challenges for dietitians working with patients whose vitamin and mineral requirements are impacted by various diseases, conditions, and medical treatment. Dietitians are the best-positioned health professionals to lead an evidence-based approach toward recommending vitamin and mineral supplements. The dietitians at the Toronto Rehabilitation Institute were authorized through a medical directive to prescribe multiple vitamins with minerals and to discontinue orders for unnecessary vitamin supplements. This is an ongoing, advanced practice initiative that focuses on the clinical efficacy for and safety of supplementation with multiple vitamins with minerals. It involves assessing the strength of evidence as it emerges in the literature, determining its relevance to specific patient populations in the practice setting and re-evaluating clinical practices for potential applications. When dietitians assume advanced practice initiatives, they are better equipped to deliver high-quality patient care. Simultaneously, state-of-the-art dietetic practice heightens dietitian recognition as a valuable member of the health care team. PMID- 15127065 TI - On the front line: practice satisfactions and challenges experienced by dietetics and nutrition professionals working in community settings in New York State. AB - The goal of this study was to understand dietetics and nutrition professionals' experiences of their practice roles. Qualitative interviews using a grounded theory design covered practitioners' perceptions of their professional roles, role enactment, and practice context. Twenty-four dietetics and nutrition practitioners varying in their work settings, length of professional experience, education, and community type were recruited through professional contacts in New York State. Interview transcripts were analyzed using the constant comparative method of qualitative data analysis. An ecological model of practice in context emerged in which participants described daily practice satisfactions and challenges arising out of interactions among their personal characteristics, client characteristics, the work setting, and the food and nutrition and health care systems. Practice satisfactions related to positive interactions and measurable outcomes of work with clients and coworkers, recognition for expert and helper roles, and involvement in disease prevention. Practice challenges centered on others' misunderstandings of the dietary change process, assessment of practice outcomes, others' respect for expertise, keeping up-to-date, client and coworker expectations, isolation from peers, and the food environment. An ecological model of dietetics and nutrition practice as experienced in community settings draws attention to the need to address challenges in the multiple contexts that frame that practice. PMID- 15127066 TI - Coordinated school health program and dietetics professionals: partners in promoting healthful eating. AB - Although research indicates that school meal programs contribute to improved academic performance and healthier eating behaviors for students who participate, fewer than 60% of students choose the National School Lunch Program or School Breakfast Program. School meal programs have a difficult time competing with foods that are marketed to young people through sophisticated advertising campaigns. Youth's preferences for fast foods, soft drinks, and salty snacks; mixed messages sent by school personnel; school food preparation and serving space limitations; inadequate meal periods; and lack of education standards for school foodservice directors challenge school meal programs as well. A coordinated school health program offers a framework for meeting these challenges and provides children and adolescents with the knowledge and skills necessary for healthful eating. This article identifies challenges facing school foodservice directors in delivering healthful meals and acquaints dietetics professionals with the coordinated school health program to be used as a tool for addressing unhealthful weight gain and promoting healthful eating. PMID- 15127067 TI - Food safety education: health professionals' knowledge and assessment of WIC client needs. AB - A written questionnaire was used to assess the opportunities and challenges for food safety education in the Special Supplemental Nutrition Program for Women, Infants, and Children (WIC). The sample included directors and health professionals with nutrition counseling responsibilities in 79 WIC clinics in a midwestern state. Seventy-two percent of the clinics participated in the study. Data analysis included descriptive statistics, chi(2) analysis, and Cronbach's alpha. Seventy-two percent reported offering food safety advice to 20% or more of their clients daily. Ninety percent identified the food-safety knowledge of their WIC clients to be fair to very poor. The need for food safety handouts, targeted for WIC clients, was identified by 27% of the respondents. WIC professionals are promoting safe food handling among a high-risk population group. Expansion of current food-safety education efforts with WIC clients is encouraged with emphasis on methods to effectively overcome barriers to safe food handling. PMID- 15127068 TI - Accuracy of estimation of large food portions. AB - To examine accuracy of reporting of large food portions, 50 adults were invited to eat a restaurant meal that included a hamburger patty, french fries, and ice cream. After eating, participants were asked to estimate the amount of food consumed using commonly used and larger-sized food models as a guide. With the use of commonly used models, underreporting was evident for all three foods. For example, the amount of hamburger patty consumed was on average 4.01 oz, although the average amount reported was 3.04 oz (P<.05). When larger-sized models were used, the amount reported more closely matched the amount consumed, although underreporting persisted for the hamburger patty and french fries. In summary, significant underreporting of large food portions may result when commonly used food models are used to assist individuals in reporting food portions. PMID- 15127069 TI - Diabetes self-management training program in a community clinic improves patient outcomes at modest cost. AB - Stringent glycemic control reduces complications and health care costs for people with diabetes. This study assessed the effectiveness of a diabetes self management training (DSMT) program at a community clinic. Education and a glucometer were provided to 70 people with type 2 diabetes in a 4-hour class, followed by individual dietitian consults and monthly support meetings. Most participants were Hispanic or African American with mean age of 49+/-10 years and mean body mass index of 34+/-9. Body weight, glycosylated hemoglobin (A1C), medications, and follow-up attendance were monitored. After 2 to 12 months of program participation, mean A1C improved from 9.7+/-2.4% to 8.2+/-2.0% (P<.001); 61% experienced positive medication outcomes. The cost of community clinic DSMT was approximately $280 per person per year, $185 for each point reduction in A1C. This study indicated that community clinic DSMT can improve glycemic control at modest cost. PMID- 15127070 TI - Vitamins A and C, calcium, fruit, and dairy products are limited in food pantries. AB - Food pantries serve over 19 million Americans, yet little is known about the nutritional quality of foods distributed in pantry bags. Foods in bags from 133 clients from 19 pantry sites were itemized, and a mean site value for nutrient and food group content was calculated. If an individual consumed the pantry foods according to the Food Guide Pyramid, the bag would contain sufficient bread group foods to last approximately 7 days; vegetable and meat/protein group foods would last about 5 days, and fruit and milk group foods would last only approximately 3 days. Foods distributed were of adequate or high nutrient density for protein, fiber, iron, and folate, but were of low nutrient density for calcium, vitamin A, and vitamin C. Creative efforts are needed for pantries to procure, store, and distribute additional fruit, dairy products, and other sources of vitamins A and C and calcium. PMID- 15127072 TI - Promoting healthful diets from a public perspective. PMID- 15127071 TI - Position of the American Dietetic Association: Functional foods. AB - It is the position of the American Dietetic Association that functional foods, including whole foods and fortified, enriched, or enhanced foods, have a potentially beneficial effect on health when consumed as part of a varied diet on a regular basis, at effective levels. The Association supports research to define further the health benefits and risks of individual functional foods and their physiologically active components. Dietetics professionals will continue to work with the food industry, the government, the scientific community, and the media to ensure that the public has accurate information regarding this emerging area of food and nutrition science. Knowledge of the role of physiologically active food components, from both phytochemicals and zoochemicals, has changed the role of diet in health. Functional foods have evolved as food and nutrition science has advanced beyond the treatment of deficiency syndromes to reduction of disease risk. This position reviews the definition of functional foods, their regulation, and the scientific evidence supporting this emerging area of food and nutrition. Foods can no longer be evaluated only in terms of macronutrient and micronutrient content alone. Analyzing the content of other physiologically active components and evaluating their role in health promotion will be necessary. The availability of health-promoting functional foods in the US diet has the potential to help ensure a healthier population. However, each functional food should be evaluated on the basis of scientific evidence to ensure appropriate integration into a varied diet. PMID- 15127073 TI - At what age should an overweight child follow a calorie-restricted diet? PMID- 15127078 TI - Association between catechol-O-methyltransferase functional polymorphism and male suicide completers. AB - Suicide has been suggested to involve catecholaminergic dysfunction and to be related to genetics. Catechol-O-methyltransferase (COMT) 158Val/Met polymorphism (GenBank Accession No. Z26491) is a polymorphism of the gene encoding COMT, a major enzyme in catecholamine inactivation. The COMT 158Val/Met polymorphism affects COMT activity, that is, the alleles encoding Val and Met are associated with relatively high and relatively low COMT activity, respectively. In this study, we hypothesized that the COMT 158Val/Met polymorphism is associated with suicide. The study population consisted of 163 suicide completers (112 males and 51 females). We found that the genotype distribution of the COMT 158Val/Met polymorphism was significantly different between male suicide completers and male controls (p=0.036), while the frequency of the Val/Val genotype, a high-activity COMT genotype, was significantly less in male suicide completers than in male controls (OR: 0.52; 95% CL: 0.31-0.89; p=0.016). However, this was not the case in females. Our results suggest that the Val/Val genotype is a protective factor against suicide in males. PMID- 15127079 TI - Improvements in neurocognitive function and mood following adjunctive treatment with mifepristone (RU-486) in bipolar disorder. AB - High cortisol levels are found in severe mood disorders, particularly bipolar disorder. Hypercortisolaemia may cause or exacerbate both neurocognitive impairment and depressive symptoms. We hypothesized that antiglucocorticoid treatments, particularly corticosteroid receptor antagonists, would improve neurocognitive functioning and attenuate depressive symptoms in this disorder. To test this hypothesis, 20 bipolar patients were treated with 600 mg/day of the corticosteroid receptor antagonist mifepristone (RU-486) or placebo for 1 week in a double-blind crossover design. Over the total 6 weeks of the study, neurocognitive and neuroendocrine function were evaluated at baseline, days 21 and 42. Mood symptoms were evaluated weekly. Nineteen subjects completed the protocol; there were no drop-outs due to adverse events. Following treatment with mifepristone, selective improvement in neurocognitive functioning was observed. Spatial working memory performance was significantly improved compared to placebo (19.8% improvement over placebo). Measures of verbal fluency and spatial recognition memory were also improved after mifepristone. Beneficial effects on mood were found; Hamilton Depression Rating Scale scores were significantly reduced compared to baseline (mean reduction of 5.1 points) as were Montgomery Asberg Depression Rating Scale scores (mean reduction of 6.05 points). No significant change occurred after placebo. These data require replication but provide preliminary evidence that glucocorticoid receptor antagonists may have useful cognitive-enhancing and possibly antidepressant properties in bipolar disorder. PMID- 15127080 TI - Preferential 5-HT1A autoreceptor occupancy by pindolol is attenuated in depressed patients: effect of treatment or an endophenotype of depression? AB - Using positron emission tomography and the selective 5-HT1A receptor radioligand [11C]WAY100635, we previously demonstrated a preferential occupancy of 5-HT1A autoreceptors, compared to postsynaptic receptors by pindolol in healthy volunteers. We have speculated that preferential occupancy may be clinically important for the purported actions of pindolol in accelerating the antidepressant effects of selective serotonin re-uptake inhibitors (SSRIs). In this study, we have examined the preferential occupancy by pindolol of 5-HT1A autoreceptors, following three different pindolol regimes (10 mg single dose, 2.5 mg t.i.d., and 5 mg t.i.d., in 15 depressed patients on SSRIs. In addition, seven healthy volunteers were examined following a single 10 mg dose of pindolol. We found a preferential occupancy of 22.6+/-7.7% following a single dose of 10 mg of pindolol, in the healthy volunteers, which was attenuated in depressed patients on the same dose of pindolol to 2.9+/-10.8% (Student's t=3.94, df=12, p=0.002). In addition, we found a significant negative correlation between the degree of preferential occupancy and the severity of depression as assessed by the Hamilton depression rating score (HAM-D), Spearman's rho=-0.728, N=14, p=0.003, in the depressed sample. A possible mechanism underlying preferential occupancy and the attenuation of this phenomenon in depressed patients on SSRIs may include changes in the proportion of high affinity 5-HT1A sites in the autoreceptor region of the midbrain raphe. Speculatively, the degree of preferential occupancy may serve as a surrogate marker for depression, or the pharmacological effects of antidepressants. PMID- 15127081 TI - Dissociable roles for the dorsal and median raphe in the facilitatory effect of 5 HT1A receptor stimulation upon cocaine-induced locomotion and sensitization. AB - A distinct role for serotonin transmission from the dorsal and median raphe nuclei (DRN and MRN, respectively) was identified in regulating the behavioral and neurochemical effects of acute and repeated cocaine administration. Serotonin 1A (5-hydroxytryptophan (5-HT)1A) receptors were stimulated by intraraphe microinjection of 8-hydroxy-2-(di-n-propylamino)tetralin (DPAT; 5 or 10 microg) and behavior, as well as extracellular neurotransmitter content in the nucleus accumbens was measured. Pretreatment of the DRN with DPAT caused a sensitization like potentiation of acute cocaine-induced motor activity and an elevation in extracellular dopamine and glutamate. In contrast, DPAT microinjection into the MRN did not alter acute cocaine-induced motor activity or extracellular levels of dopamine or glutamate. Acutely, DPAT microinjection into either raphe nucleus reduced the basal and acute cocaine-stimulated levels of extracellular serotonin. Pretreatment with DPAT before systemic cocaine administration was continued for 5 days, and 3 weeks after the last injection, all rats were administered a cocaine challenge injection. The sensitized behavioral and neurochemical response produced by repeated cocaine in control subjects was unaffected by the intra-DRN administration of DPAT. However, in animals administered DPAT into the MRN, both the sensitized motor response and the increase in glutamate were augmented, while the sensitized serotonin response was blocked, without altering dopamine sensitization. These data show a differential role for 5-HT1A receptors in the DRN and MRN in the acute and sensitized effects of cocaine. While the DRN is involved in the acute effects of cocaine, neuroadaptations in the MRN may regulate the long-term consequences of repeated cocaine exposure. PMID- 15127082 TI - Comparison of the effects of citalopram and escitalopram on 5-Ht-mediated neuroendocrine responses. AB - Acute oral administration of selective serotonin re-uptake inhibitors (SSRIs) increases plasma cortisol by facilitating brain serotonin activity. Recently, salivary cortisol sampling has grown in popularity as a noninvasive means of assessing HPA axis activity. The aim of the present study was to find out whether acute oral administration of the SSRI, citalopram, increases salivary cortisol in healthy volunteers and whether the increase produced by an equivalent dose of its active isomer, escitalopram, is greater. A total of 15 healthy subjects were tested on three occasions receiving either oral citalopram (20 mg), escitalopram (10 mg), or placebo in a double-blind, randomized, crossover design. Salivary cortisol and plasma cortisol and prolactin were measured for 240 min after each treatment. Relative to placebo, both citalopram and escitalopram increased salivary and plasma cortisol levels with no evidence of consistent differences between them. Plasma prolactin concentration was not altered by either active treatment. Plasma and salivary cortisol responses after citalopram but not escitalopram correlated significantly. The present study does not support an enhanced effect of escitalopram on 5-HT-mediated neuroendocrine responses. PMID- 15127083 TI - Effects of lorazepam on the neuromagnetic mismatch negativity (MMNm) and auditory evoked field component N100m. AB - The mismatch negativity (MMN) as an auditory evoked potential is thought to reflect an early, preconscious attention process. While this component has gained great importance in studies on clinical populations and in basic research on auditory information processing, the involvement of different neurotransmitters in the generation of this component is less well understood. We investigated the impact of the benzodiazepine lorazepam as a GABA agonist on the neuromagnetic MMN (MMNm) and auditory evoked field component N100m. A group of 12 healthy subjects was studied in single blind trials under the following three conditions: after the intake of 1.25 mg lorazepam, 100 mg caffeine or placebo. Neuromagnetic recordings were obtained before drug intake and three times after it. Controlled visual attention was tested additionally using a version of the Continuous Performance Test (CPT). The neuromagnetic activity was reconstructed by a single moving dipole, and the dipole moment and its latency were compared between conditions and time points of measurement. Lorazepam diminished the signal detection performance in the CPT 25 min after drug intake. The source of the field component N100m was attenuated, most significantly in the recording 105 min after lorazepam intake. The attenuation of the MMNm under lorazepam became significant at 105 min, but was visually less apparent, because in all conditions a decrease of the MMNm dipole moment within the course of a session was observed. Besides the already known effects of benzodiazepines on controlled attention functions, preconscious attention functions as reflected in the MMN are impaired by acute benzodiazepine intake. MMN studies on clinical populations have to be controlled for the recording time because of the strong habituation of this component. PMID- 15127084 TI - The serotonin 5-HT2A receptors antagonist M100907 prevents impairment in attentional performance by NMDA receptor blockade in the rat prefrontal cortex. AB - We investigated whether 5-HT2A receptors contribute to the control of attentional performance by glutamate NMDA receptor mechanisms in the medial prefrontal cortex (mPFC). We examined the effects of NMDA receptor blockade in the mPFC on attentional performance by infusing a competitive glutamate NMDA receptor antagonist, 3-(R)-2-carboxypiperazin-4-propyl-1-phosphonic acid (CPP) into the mPFC of rats performing a task of divided and sustained visual attention. The five-choice serial reaction time task provides indices of attentional functioning (% correct responses), executive control (measured by anticipatory and perseverative responses) and speed. A dose of 10 ng CPP injected bilaterally into the mPFC increased anticipatory and perseverative responding; 50 ng reduced accuracy. Increasing the stimulus duration alleviated the CPP-induced accuracy deficit but did not reduce its effects on anticipatory and perseverative responses. CPP at 50 ng caused motor hyperactivity whereas lower doses had no effect. [R-(+)-(2,3-dimethoxyphenyl)-1-[2-(4-fluorophenylethyl)]-4-piperidine methanol] (M100907) (M100907), a 5-HT2A receptor antagonist, injected subcutaneously at 10 and 40 microg/kg, had no effect on accuracy but dose dependently reversed the impairment induced by 50 ng CPP. Both doses of M100907 completely abolished CPP-induced anticipatory but not perseverative over responding. At the dose of 40 microg/kg M100907 reversed CPP-induced motor hyperactivity. This study provides evidence that the prefronto-cortical glutamate NMDA system may make an important contribution to the control of attention and executive functions. It also indicates that 5-HT2A receptors may serve to optimize attentional selectivity and improve some aspects of executive control. PMID- 15127085 TI - The Effects of dizocilpine and phencyclidine on prepulse inhibition of the acoustic startle reflex and on prepulse-elicited reactivity in C57BL6 mice. AB - Prepulse inhibition (PPI) of the acoustic startle response refers to the reduction in startle reaction to a startle-eliciting stimulus when it is shortly preceded by a subthreshold prepulse stimulus. Here, we evaluated the possible effects on prepulse-elicited reactivity by dizocilpine (MK-801) and phencyclidine (PCP) in the PPI of acoustic startle paradigm in C57BL6/J mice. The aim was to ascertain whether these two drugs would affect prepulse-elicited reactivity in a manner similar to apomorphine, which enhances prepulse-elicited reactivity at doses that disrupt PPI. In two dose-response studies, we showed that both drugs exhibited a tendency to attenuate prepulse-elicited reaction at higher doses when PPI was severely disrupted. On the other hand, at lower doses when PPI was marginally disrupted, reaction to the prepulse, if anything, tended to increase. It is concluded that PPI disruption induced by noncompetitive NMDA receptor antagonists can be distinguished from apomorphine-induced PPI disruption by their concomitant effects on prepulse-elicited reactivity. Our data support the suggestion that dopamine receptor agonists and NMDA receptor antagonists disrupt PPI via interference with distinct neural pathways or neuronal systems. PMID- 15127086 TI - Electrophysiological evidence for the tonic activation of 5-HT(1A) autoreceptors in the rat dorsal raphe nucleus. AB - Serotonin (5-hydroxytryptamine, 5-HT) and norepinephine (NE) neurons have reciprocal connections. These may thus interfere with anticipated effects of selective pharmacological agents targeting these neurons. The main goal of the present study was to assess whether the somatodendritic 5-HT(1A) autoreceptor is tonically activated by endogenous 5-HT in anesthetised rats, using in vivo extracellular unitary recordings. In rats with their NE neurons lesioned using 6 hydroxydopamine (6-OHDA) and in controls administered the NE reuptake inhibitor desipramine to suppress NE neuronal firing, the alpha2-adrenoceptor agonist clonidine no longer inhibited 5-HT neuron firing, therefore indicating the important modulation of the firing activity of 5-HT neurons by NE neurons. In control rats, the administration of the potent and selective 5-HT(1A) receptor antagonist WAY 100,635 ((N-[2-[4(2-methoxyphenyl)-1-piperazinyl]ethy]-N-(2 pyridinyl)cyclohexanecarboxamide trihydroxychloride) (100 microg/kg, i.v.) did not modify the spontaneous firing activity of 5-HT neurons, but in NE-lesioned rats using either 6-OHDA or DSP-4, WAY 100,635 produced a mean firing increase of 80 and 69%, respectively. When desipramine and D-amphetamine were used in control rats to prevent alterations in the availability of NE in the dorsal raphe, again WAY 100,635 produced a significant disinhibition of the firing of 5-HT neurons (83 and 53%, respectively). These data support the notion that the NE system tonically activates the firing activity of 5-HT neurons. When the fluctuations of the function of NE neurons normally produced by WAY 100,635 were prevented, a tonic activation of 5-HT(1A) autoreceptors by endogenous 5-HT was unmasked. PMID- 15127087 TI - Fitness advantage and cytoplasmic incompatibility in Wolbachia single- and superinfected Aedes albopictus. AB - Wolbachia are obligate, maternally inherited, intracellular bacteria that infect numerous insects and other invertebrates. Wolbachia infections have evolved multiple mechanisms to manipulate host reproduction and facilitate invasion of naive host populations. One such mechanism is cytoplasmic incompatibility (CI) that occurs in many insect species, including Aedes albopictus (Asian tiger mosquito). The multiple Wolbachia infections that occur naturally in A. albopictus make this mosquito a useful system in which to study CI. Here, experiments employ mosquito strains that have been introgressed to provide genetically similar strains that harbor differing Wolbachia infection types. Cytoplasmic incompatibility levels, host longevity, egg hatch rates, and fecundity are examined. Crossing results demonstrate a pattern of additive unidirectional cytoplasmic incompatibility. Furthermore, relative to uninfected females, infected females are at a reproductive advantage due to both cytoplasmic incompatibility and a fitness increase associated with Wolbachia infection. In contrast, no fitness difference was observed in comparisons of single- and superinfected females. We discuss the observed results in regard to the evolution of the Wolbachia/A. albopictus symbiosis and the observed pattern of Wolbachia infection in natural populations. PMID- 15127088 TI - Comparative genomics: mining the crucial 1%. PMID- 15127089 TI - Plasma and lung macrophage responsiveness to carotenoid supplementation and ozone exposure in humans. AB - OBJECTIVE: To examine the effect of ozone exposure and vegetable juice supplementation on plasma and lung macrophage concentrations of carotenoids. DESIGN: A randomized trial. SETTING: Subjects were exposed to ambient air prior to antioxidant supplementation and to ozone after antioxidant supplementation or placebo. Exposures occurred while exercising intermittently in a controlled metabolic chamber at the Human Studies Division, US EPA. SUBJECTS: In all, 23 healthy subjects between ages of 18 and 35 y. INTERVENTIONS: Subjects consumed a low fruit and vegetable diet for 3 weeks. After the first week, subjects underwent a sham exposure to filtered air with exercise, followed by bronchoalveolar lavage (BAL). Subjects were randomly assigned into supplement (one can vegetable juice, vitamins C and E daily) or placebo (orange soda, placebo pill daily) groups for 2 weeks. After the 2-week intervention, subjects were exposed to 0.4 ppm (784 microg/m(3)) ozone for 2 h with exercise followed by BAL. Blood samples were drawn before, immediately after and 3 h postexposure on each exposure day. The concentrations of nine carotenoids were determined by HPLC in BAL macrophages and plasma samples. RESULTS: Plasma concentrations of all the carotenoids that were present in the vegetable juice (except cis-beta-carotene) increased significantly in the supplemented group. Lung macrophage alpha-carotene concentrations increased significantly, lycopene isomers increased slightly, and all other carotenoids decreased (nonsignificantly) in the supplementation group following the intervention. Ozone exposure resulted in decreases in several carotenoids in plasma of the placebo group, but not in the supplemented group. CONCLUSIONS: Lung macrophage concentrations of carotenoids can be manipulated by diet. Ozone is a potent environmental oxidant that appears to reduce plasma carotenoids in nonsupplemented individuals. PMID- 15127090 TI - Does vitamin C supplementation influence the levels of circulating oxidized LDL, sICAM-1, sVCAM-1 and vWF-antigen in healthy male smokers? AB - OBJECTIVE: To examine the effects of vitamin C supplementation on the concentration of oxidation markers, in particular, circulating oxidized LDL (OxLDL) and on endothelial activation markers. DESIGN: Randomized double-blind, placebo-controlled crossover trial. SETTING: Belgian population of the city of Leuven. SUBJECTS: A total of 34 healthy male smokers aged 26-73 y. INTERVENTION: Smokers were randomly assigned to receive either vitamin C (250 mg twice daily) or placebo capsules, each to be taken for 4 weeks. After a 1-week washout period, participants then crossed over to the alternative capsules for further 4 weeks. MEAN OUTCOME MEASURES: Markers of oxidation (bilirubin, uric acid, alpha tocopherol, retinol, malondialdehyde, circulating Oxidized LDL (OxLDL)) and markers of endothelial activation (sICAM-1, sVCAM-1, vWF-antigen) were analysed. RESULTS: Plasma ascorbate concentrations significantly increased from 46.6+/-17.6 to 70.1+/-21.2 mumol/l after a 4-week treatment with 500 mg vitamin C per day. The other plasma antioxidants concentrations, including bilirubin, uric acid, alpha-tocopherol and retinol, were similar in both treatment periods. Vitamin C did not change plasma malondialdehyde and circulating OxLDL compared with placebo (vitamin C 0.73+/-0.25 mg/dl OxLDL; placebo 0.72+/-0.21 mg/dl OxLDL). After vitamin C supplementation, neither sICAM-1 and sVCAM-1 levels nor the concentration of vWF-antigen significantly differed from placebo condition. CONCLUSIONS: Oral supplementation of vitamin C is not associated with changes in markers of oxidation or endothelial activation in healthy male smokers. PMID- 15127091 TI - Physical activity pattern of children assessed by triaxial accelerometry. AB - OBJECTIVES: Accelerometry was used to assess the relationship between the physical activity level (PAL) and time spent on activities of various intensities in children. DESIGN: A total of 20 children aged 8.6+/-3.3 y wore a triaxial accelerometer (Tracmor2) for 2 weeks. PAL was calculated with Tracmor2 output data. The fraction of time spent on activities with a given level of intensity (low, moderate, high) was calculated. The fractions of time spent on activities of different intensities were compared with previously obtained data for young adults and elderly persons. RESULTS: PAL showed an inverse relation with the percentage of time spent on low-intensity activities (r = -0.76; P < 0.0001) and a positive relation with the percentage of time spent on high-intensity activities (r = 0.93; P < 0.0001). The fraction of time spent on low-intensity activities was smaller in children than in young adults (P < 0.05) and elderly persons (P < 0.0001), while the fraction spent on high-intensity activities (P < 0.0001) was larger. CONCLUSIONS: The present data are important for a better understanding of physical activity in children, which is necessary for education and prevention about physical (in)activity in childhood. Our observations suggest that to obtain a higher PAL in children, they should be given the opportunities to perform high-intensity activities. PMID- 15127092 TI - Correlation between dietary glycemic index and cardiovascular disease risk factors among Japanese women. AB - OBJECTIVE: To examine the correlation between dietary glycemic index (GI) and cardiovascular disease (CVD) risk factors among subjects who consume white rice as a staple food. DESIGN: A cross-sectional study was conducted to explore the associations between dietary GI, dietary glycemic load (GL) and dietary intakes, and CVD risk factors. Dietary GI and GL were calculated from a 3-day (including two consecutive weekdays and one holiday) dietary records. SETTING: A weight reduction program at a municipal health center in Tokyo, Japan. SUBJECTS: A total of 32 women aged 52.5+/-7.2 y participated in the weight-reduction program. RESULT: The GI food list made for the current study calculated for 91% of carbohydrate intakes measured. The mean dietary GI was 64+/-6, and the mean dietary GL was 150+/-37. Individuals in the highest tertile of GI consumed more carbohydrate, mostly from white rice (P<0.001), and less fat (P<0.01). Individuals in all three groups by tertile of GL showed similar tendencies. In the lowest GI tertile, the highest concentration of HDL-cholesterol and lowest concentration of triacylglycerol and immunoreactive insulin were observed (P<0.01). In the lowest GL tertile, the highest concentration of HDL-cholesterol and the lowest concentration of triacylglycerol were observed (P<0.05). CONCLUSION: Calculated dietary GI and GL were positively associated with CVD risk factors among the Japanese women who consumed white rice as a staple food. PMID- 15127120 TI - [Induced-division of neurons derived from neural stem cells]. AB - In order to explore if mature neurons derived from neural stem cells have the potentiality to divide, we utilized the chemical digestion method to disperse the adult rat brain tissue into single cells, and culture them in serum-free medium. After being cultured for about eight days in vitro, the neural stem cells were induced to differentiate into neurons. The neurons were further induced to divide. Utilizing the method of serial photograph and NF-160 immunocytochemistry, the processes of division of some neurons were recorded. At the same time, PCNA+NF-160 (or Chat, GABA, GAD) double label were used to investigate if the dividing-neurons were mature ones. After the neural stem cells were induced to differentiate in vitro for eight days, they possessed the shape and character of mature neurons. The differentiated neuron had a big nucleus and one or two distinct nucleolus in the nuclear. Within the perikaryon,there were a large amount of dense and Nissl body-like structure. Several long processes emerged from various locations of the cell body. Then, EGF and bFGF were added into the medium to induce division. After two days of induced-division, neuron-like cells were observed to divide; moreover, the number of neuron-like cells in the region increased continually. Immunocytochemistry demonstrated these cells were NF-160 positive. Serial photographs of dividing-process of neuron-like cells were obtained and their daughter cells were also NF-160-positive. After PCNA+NF-160 (or Chat, GABA, GAD) double label, some cells showed brown cell plasma and black nucleus. The above-mentioned results indicate that neurons, which were previously thought to be end-differentiated, can be re-called into cell cycle under appropriate conditions. Mature neurons still have the potential to divide, proliferate and self-renew. PMID- 15127119 TI - Structural survey of airway sensory receptors in the rabbit using confocal microscopy. AB - Information on the morphology of airway receptors is limited. The present study surveys rabbit airway receptors using immunohistochemical and fluorescent labeling to identify their structure with confocal microscopy. Various receptor types were observed to have multiple branches where a parent axon fed several structures. Receptors were located in different layers of the airway, i.e., smooth muscle, lamina propria (submucosa) and the epithelium. Smooth muscle and submucosal receptors were innervated by thick myelinated fibers, while epithelial receptors were supplied by thin-diameter axons. Structures of smooth muscle receptors and some submucosal receptors covered a relatively large area, while epithelial receptors were less extended. In addition, intrapulmonary ganglia were also labeled. Some were closely associated with the axons of smooth muscle receptors. PMID- 15127121 TI - Temporal and spatial distribution of VIP, CGRP and their receptors in the development of airway hyperresponsiveness in the lungs. AB - To explore the role of intrapulmonary neuropeptides in the development of airway hyperresponsiveness, we established an animal model of airway hyperresponsiveness (AHR) in rabbits by using ozone exposure. With the model, after test of the mechanics of respiration and bronchoalveolar lavage assay, the levels of vasoactive intestinal peptide (VIP) and calcitonin gene-related peptide (CGRP) in the lungs were determined by radioimmunoassay, and the expression of mRNA coding receptors of these two neuropeptides was evaluated by reverse transcriptional polymerase chain reaction (RT-PCR). At the same time, the distribution of VIP receptor-1 (VIPR1) and CGRP receptor-1 (CGRPR1) in lung tissues and its time course were examined by in situ hybridization. The results showed: (1) in ozone stressing groups, airway resistance increased significantly and typical inflammatory pathological changes were observed in pulmonary tissue slides, including neutrophil and eosinophil infiltration, mucus exudation and bronchial epithelial cells (BECs) shedding; (2) with elongation of ozone exposure, the levels of VIP and CGRP in the lungs increased at first, reaching a peak on d 2 to 4, then decreased slowly, and CGRP peaked somewhat earlier than VIP; (3) mRNA expression of the two neuropeptide receptors in the lungs changed in a similar manner like VIP and CGRP, but the high level of mRNA expression of VIPR1 lasted longer than that of CGRPR1; and (4) in situ hybridization for neuropeptide receptors demonstrated that, in unstressed control, VIPR1 and CGRPR1 positive cells appeared in the airway epithelium, pulmonary interstitial and focal areas of airway and vascular smooth muscles. With the elongation of ozone exposure, hybridization stained deeper and the majority of positive cells were located around the vessels and bronchus except a few in the alveoli. At 8 d, only a small number of positive cells were seen in the lungs. From the results, it is concluded that ozone-stressing can induce the development of AHR, in which VIP and CGRP may play important roles. That implies, through binding to CGRPR1, CGRP stimulates an early inflammation response which contributes in cleaning up of irritants, while VIP exerts a later dampening of pulmonary inflammation response. These two neuropeptides may play sequential and complementary roles in the development of AHR. PMID- 15127122 TI - Ischemia-induced release of cytochrome c from mitochondria and up-regulation of Bcl-2 expression in rat hippocampus. AB - To evaluate the effects of different antagonists on the release of cytochrome c from mitochondria to cytosol and the expression of Bcl-2 in mitochondria in rat hippocampus after ischemia, we examined Bcl-2 and cytochrome c expression by immunoblotting using 4-vessel occlusion (4-VO) as brain ischemia model. The results showed that after 24 h ischemia/reperfusion (I/R) cytochrome c decreased markedly in mitochondria, which was correspondingly increased in the cytosolic fraction. Bcl-2 expression was time-dependent, reaching its peak level after 6 h I/R. In all those samples, there were no alterations in the subcellular distribution of cytochrome oxidase, a mitochondrial respiratory chain protein. The decreases in Bcl-2 and cytochrome c in mitochondria were restored by pretreatment with non-competitive NMDA receptor antagonist ketamine or L-type voltage-gated Ca(2+) channel (L-VGCC) antagonist nifedipine at 20 min prior to ischemia. The results demonstrate that the release of cytochrome c from mitochondria to cytosol and the up-regulation of Bcl-2 are possibly mediated by NMDA receptors or L-VGCC following brain ischemia. Cytochrome c release may be injurious while Bcl-2 up-regulation may be protective to ischemic hippocampus. PMID- 15127124 TI - [Effect of acetazolamide on the latency of hyperbaric oxygen-induced convulsion]. AB - The purpose of the present study was to explore the relation between the modulation of cerebral blood flow and the latency of hyperbaric oxygen-induced convulsion. There were two parts in this study. First, the effect of acetazolamide on the latency of hyperbaric oxygen-induced convulsion was observed. 32 Sprague-Dawley (SD) rats were randomly divided into four groups: the acetazolamide 200, 20, 2 mg/kg body weight and normal saline (NS) group. The animals were given intraperitoneally acetazolamide or NS, respectively, before being exposed to the pressure of 6 ATA (absolute atmosphere) of pure oxygen. The time from exposure to the onset of seizure (clonic-tonic convulsion) was recorded for each animal according to behavioral observation. Second, the changes in maleic dialdehyde (MDA) and the activity of glutathione peroxidase (GSH-PX) were measured after acetazolamide treatment. 40 SD rats were randomly divided into five groups: NS group, 6 min with NS group, 6 min with acetazolamide group, 16 min with NS group, and 16 min with acetazolamide group. The dose of acetazolamide was 20 mg/kg body weight. After injection of NS or acetazolamide, the animals were subjected to the pressure of 6 ATA of pure oxygen in respect to its time course group. The rats were decapitated and the cortex, hippocampus, and striatum of brains were dissected and homogenized. The content of MDA and the activity of GSH-PX in these tissues were determined. We found that (1) there was a significant difference in the latency of hyperbaric oxygen-induced convulsion between the acetazolamide 200 mg/kg group and the NS control group, as well as between the acetazolamide 20 mg/kg group and the NS control group (P<0.01), whereas there was no significant difference between the NS group and the acetazolamide 2 mg/kg weight group (P>0.05). The latency of these groups were listed as follows: 9.78+/-1.94 min for 200 mg/kg body weight group, 10.92+/-1.68 min for 20 mg/kg body weight group, 24.32+/-4.33 min for 2 mg/kg body weight group and 22.02+/-4.32 min for NS control group. (2) there was no significant difference between all groups in the activity of GSH-PX, though it varied with the oxidation levels. In the cortex and hippocampus, the activity of GSH-PX boosted up at first, but with the progress of the oxidation it was impaired. In the striatum, the activity of GSH-PX increased stepwise with the aggravation of the oxidation. The MDA content in the cortex increased significantly in the group of 6 min with acetazolamide (P<0.01), as well as the group of 16 min with acetazolamide group both in cortex and hippocampus (P<0.01, P<0.05). The MDA content of all groups is correlated with the dose of acetazolamide and the exposure time. These results suggest that acetazolamide which dilates the brain arteriolar obviously shortens the latency of hyperbaric oxygen-induced convulsion, and that acetazolamide dilates the vessels and increases the supply of the oxygen breaking into the brain tissues and aggravates the oxidation. The hyperbaric oxygen-induced convulsion correlates closely with the oxidation injury. PMID- 15127123 TI - [Expression of transient receptor potential channel 4 in striatum and hippocampus of rats is increased after focal cerebral ischemia]. AB - This paper was designed in middle cerebral artery occlusion (MCAO) model of rats, to explore the role of transient receptor potential channel 4 (TRPC4) as Ca(2+) selective channel by detecting the changes of the expression of TRPC4 in different parts of cerebral tissues under the condition of focal cerebral ischemia. The rats were sacrificed after MCAO surviving time 6 h, 12 h, 1 d, 3 d. As determined by Western blot, the expressions of TRPC4 in striatum and hippocampus of 12 h, 1 d, 3 d groups were significant higher than that in the control group (P<0.05). Immunohistochemical staining showed that the TRPC4 immunoreactive substances were present in the membrane of neurons. Compared with the control group, immunostaining positive cells increased in hippocampus and striatum of cerebral ischemia groups. The TRPC4 immunostaining positive cells increased significantly in 1d-group and 3d-group (P<0.05). It suggests that as a Ca(2+) selective channel, the variance of the expression of TRPC4 may play a role in acute and delayed neuronal injury in focal cerebral ischemia. PMID- 15127125 TI - Dual action of lysophosphatidic acid in cultured cortical neurons: survival and apoptogenic. AB - The effect of lysophosphatidic acid (LPA), with a wide range of its different concentrations, upon cultured mouse cortical neurons was assessed by electrophoresis of DNA fragments, HO33342 and TUNEL stainings, and also by ultrastructural examination at times. The results showed that administration of LPA at lower concentrations (0.1-30 micromol/L) dose-dependently protected cortical neurons from apoptosis that was induced by deprivation of serum from the cultural medium, while 50 micromol/L or higher concentrations of LPA failed to show this effect; and moreover, the concentrations higher than 50 micromol/L induced apoptosis in neurons cultured in serum-containing complete medium. These results suggest that a moderate concentration of LPA may play as a survival factor in apoptotic cortical neurons, while an excessive level of LPA induces apoptosis in neurons cultured in complete medium. PMID- 15127126 TI - [The expression of Smac and XIAP in rat hippocampus following limbic seizure induced by kainic acid injection into amygdaloid nucleus]. AB - To determine whether Smac/DIABLO (second mitochondrial activator of caspases/direct inhibitor of apoptosis protein-binding protein of low isoelectric point [PI]) and XIAP (X-chromosome-linked inhibitor of apoptosis protein) serve to regulate neuronal apoptosis following seizures, we investigated seizure induced changes in caspase-9, Smac/DIABLO and XIAP protein expression and the in vivo effect of caspase-9 inhibition. Animals received unilateral intra-amygdaloid injection of kainic acid (0.5 microg) to induce seizures for 1 h. The seizures were then terminated by diazepam (30 mg/kg). Animals were killed 0, 2, 4, 8, 24 or 72 h following diazepam administration. The apoptotic and surviving neurons in hippocampus were observed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and cresyl violet staining, the expression of Smac/DIABLO, XIAP and caspase-9 was detected with immunofluorescence and western blot. The results showed that the levels of XIAP and the 46-kDa proenzyme form of caspase-9 were unaffected by the seizures. The expression of Smac increased at 2 h and the 37-kD cleaved fragment of caspase-9 was detected at 4 h, TUNEL-positive neurons appeared at 8 h and reached maximal at 24 h following seizure cessation within the ipsilateral (the same side as the intra-amygdaloid injection of kainic acid) CA3 subfield of the hippocampus. Intracerebroventricular infusion of caspase-9 inhibitor z-LEHD-fluoromethyl ketone (z-LEHD-fmk) significantly decreased TUNEL-positive neurons and increased the number of surviving cells. Caspase-9 immunoreactivity increased and Smac/DIABLO, XIAP immunoreactivity became extensive within the ipsilateral CA3 neurons. TUNEL-positive neurons and the alterations of the expression of Smac/DIABLO and XIAP within the ipsilateral CA3 were not detected within the contralateral hippocampus. These results suggest that seizures lead the translocation of Smac/DIABLO into the cytosol, the activation of caspase-9 and the change of subcellular locoalization of XIAP. These changes may play a role in the brain damage induced by seizures. Caspase-9 is possibly a potential therapeutic target in the treatment of brain injury associated with seizures. PMID- 15127127 TI - Age-related changes in deterministic behaviors of nociceptive firing of rat dorsal horn neurons. AB - To demonstrate the age-related changes in the dynamics of the nociceptive discharge of dorsal horn nociceptive neurons, the nonlinear prediction method was used to quantify the degree of deterministic behavior within the interspike interval series of tissue injury-induced firing of spinal nociceptive neurons in anesthetized adult young (3-4 months) and aged (>22 months) rats. Subcutaneous bee venom injection induced long-term discharge of spinal wide dynamic range (WDR) neurons in both groups. However, the nociceptive discharge of single WDR neurons in the aged group showed higher determinism when compared with the adult young rats. This result suggests that the dynamics of single nociceptive neurons may not remain constant throughout the life span, and this age-associated change may be an underlying mechanism for various pain manifestations in the elderly population. PMID- 15127128 TI - [Alteration of gene expression profiles of cultured embryo rat cortex induced by phenylalanine]. AB - To have more insight into the mechanism of neuronal injury in phenylketonuria patients, gene expression profiles were studied in cell culture of embryonic rat cortical neurons induced by phenylalanine. Randomly chosen cortical cultures for 3 d were treated by 0.9 mmol/L phenylalanine for 12 h. Control group of the same batch was treated with the same volume of medium. Total RNA was extracted and hybridized with the Affymetrix gene chip U34 according to the protocol provided by the Affymetrix Company. Real-time PCR was used to further confirm the result. We found that the hybridization signals of 167 genes were increased among the total 1323 probes plotted on the chip. The 167 increased genes could be functionally categorized into signal transduction, neuron related, cytoskeleton, metabolism, ion channels, transcription factors, cytokines, and apoptosis related. Signals of 7 probes were decreased, which accounted to 0.5% of the total number. A series of genes that were not reported before were up-regulated by phenylalanine, including Ca(2+)/calmodulin-dependent protein kinase, Brain type II (CaMK II), Ras, P38 MAP kinase, L-voltage dependent calcium channel, some genes related to vesicle formation and transmitter release, some glutamate receptor subunits and glutamate transporters. According to the gene expression profile, it is likely that multi-processes are involved in the neuronal injury induced by high phenylalanine, such as the activation of the NMDR-Ca(2+)- CaMK II - Ras- P38 axis, the abnormality in neurotransmitter release. Our study also suggests that the excitatory neurotransmitter glutamate may play a role in the neural pathology of phenylketonuria. PMID- 15127129 TI - Injection of adenosine into the renal artery activates spontaneous activity of renal afferent nerve fibers. AB - The effects of injection of adenosine into the renal artery on multi- and single unit spontaneous discharges of renal afferent nerve fibers were investigated in anesthetized rabbits. The results obtained are as follows: (1) injection of 50, 100, and 200 nmol/kg adenosine into the renal artery increased the renal afferent nerve activity (ARNA) in a dose-dependent manner with unchanged arterial pressure; (2) pretreatment with 8-cyclopenthl-1,3-dipropylxanthine (DPCPX, 160 nmol/kg), an adenosine A1 receptor antagonist, partly abolished the effect of adenosine; and (3) pretreatment with a nitric oxide synthase inhibitor N(omega) nitro-L-arginine methylester (L-NAME, 0.1 mmol/kg) significantly enhanced the ARNA response to adenosine. The results suggest that injection of adenosine into the renal artery activates ARNA via adenosine receptors in anesthetized rabbits and that nitric oxide may be involved in regulating the activity of renal sensory nerve fibers as an inhibitory neurotransmitter. PMID- 15127130 TI - [Effect of chronic electrical stimulation of phrenic nerve at different frequencies on mRNA and protein expression of skeletal DHPR(alpha1) and RyRs in the diaphragm muscle of rabbits]. AB - The mRNA and protein expression of skeletal dihydropyridine receptor isoform alpha1 subunit (DHPR(alpha1)) and ryanodine receptor(1-3) (RyR(1-3)) during chronic electrical stimulation (CES) of phrenic nerve have rarely been explored. In the present study, we explored the signal translation mode of calcium release unit in diaphragm muscle of rabbits after CES. Thirty rabbits were used and randomly divided into the normal, 10, 20, 50 and 100 Hz groups. Phrenic nerve was continuously (5 weeks, 2x 2 h/d) stimulated at 10, 20, 50 and 100 Hz respectively (impulse width 0.2 ms, 3~6 waves/time, 45 times/min, 10~20 V). Reverse transcription PCR and immunohistochemical methods were employed. The results showed that mRNA and protein expressions of DHPR(alpha1) and RyR(1) in 10 and 20 Hz groups were more significantly lower than those in the control group (P<0.01), but mRNA and protein expressions of DHPR(alpha1) and RyR(1) were significantly higher in 50 and 100 Hz groups than those in the control group (P<0.01); a lower level of mRNA expression of RyR(2) was found in 10 and 20 Hz groups. It is suggested that the calcium release unit and the signal transduction mode between DHPR and RyRs were altered from conformational changes of linked proteins to Ca(2+)-induced Ca(2+) release (CICR) in the diaphragmatic muscle of rabbits after chronic low-frequency electrical stimulation of phrenic nerve for 5 weeks. PMID- 15127131 TI - Effects of genistein on intracellular free-calcium concentration in guinea pig ventricular myocytes. AB - The effects of genistein (GST) on intracellular calcium concentration ([Ca(2+)](i)) were investigated in guinea pig ventricular myocytes. [Ca(2+)](i) was detected by confocal microscopy and represented by relative fluorescent intensity (FI-F(0)) /FI(0), %). The results showed that GST (10-40 micromol/L) reduced [Ca(2+)](i) in normal Tyrode's solution, Ca(2+)-free Tyrode's solution and normal Tyrode's solution containing 3 mmol/L EGTA in a concentration dependent manner. The effects of GST on [Ca(2+)](i) in normal Tyrode's solution were partially inhibited by pretreatment with sodium orthovanadate, a potent inhibitor of protein tyrosine phosphatase, or L-type Ca(2+) channel agonist Bay K8644. GST also markedly inhibited the ryanodine-induced [Ca(2+)](i) responses in Ca(2+)-free Tyrode's solution. When Ca(2+) waves were produced by increasing extracellular Ca(2+) concentration from 1 to 10 mmol/L, GST (40 micromol/L) could block the propagating waves of elevated [Ca(2+)](i), and reduce the velocity and duration of propagating waves. These results suggest that GST may reduce the [Ca(2+)](i) in isolated guinea pig ventricular myocytes. The inhibition of voltage-dependent Ca(2+) channel, tyrosine kinase inhibition, and alleviation of Ca(2+) release from SR are possibly involved in the GST effect. PMID- 15127132 TI - [Comparison of changes in left ventricular gene expression profiles from different cardiac hypertrophy models in rats]. AB - To get insights into the principles of gene expression changes during cardiac hypertrophy, three rat cardiac hypertrophy models were prepared, i.e., suprarenal abdominal aortic stenosis (SRS), arterial-vein fistula (AVF) and continuous jugular vein infusion of norepinephrine (NEi). The cardiac function and structure were analyzed by echocardiograph as well as histological examination. Total RNA of left ventricles was extracted and gene expression profiles were analyzed by cDNA microarray. SRS and NEi induced concentric cardiac hypertrophy and AVF induced eccentric hypertrophy in rats, among which NEi caused obvious cardiac fibrosis. The changes of gene expression profiles were compared comprehensively across different pathologic cardiac hypertrophy models. While gene expression profiles of different cardiac hypertrophy models compared with pairs, parts of the genes involved were found overlapped, and mostly the gene expression changed in the same direction between two models, but some of them changed in the opposite directions. Expression levels of 19 genes were found changed across all cardiac hypertrophy models, and genes relatively regulated in a specific model was also found when comparison of all the three models was carried out. Novel clues for further study might derive from the results mentioned above, and some genes might be the marker genes of cardiac hypertrophy or the targets of therapy. PMID- 15127133 TI - Capsaicin facilitates carotid sinus baroreflex in anesthetized rats. AB - The effects of capsaicin (CAP) on the carotid sinus baroreflex were studied in 30 anaesthetized rats with perfused isolated carotid sinus. The results are as follows. (1) By perfusing the isolated carotid sinus with CAP (1 micromol/L), the functional curve of the baroreflex was shifted to the left and downward, with a peak slope (PS) increasing from 0.34+/-0.01 to 0.42+/-0.01 (P<0.01), whereas the reflex decrease (RD) in mean arterial pressure was enhanced from 36.51+/-1.26 to 45.01+/-0.71 mmHg (P<0.01). Meanwhile, the threshold pressure, equilibrium pressure and saturation pressure were all significantly decreased from 70.43 +/ 2.09 to 52.86 +/-2.80 mmHg (P<0.01), 95.5+/-1.71 to 87.00+/-1.58 mmHg (P<0.01) and 177.60+/-1.37 to 163.55+/-2.12 mmHg (P<0.01), respectively. Among the functional parameters of carotid baroreflex, the changes in PS and RD induced by capsaicin were dose-dependent. (2) By pretreatment with ruthenium red (RR, 100 micromol/L), an antagonist of vanilloid receptor subtype 1 (VR(1)), the above effects of CAP on carotid baroreflex were abolished. (3) The CAP-induced change in the baroreflex was also eliminated by pretreatment with glibenclamide (20 microm ol/L), a K(ATP) channel blocker. On the basis of the results, it is concluded that CAP facilitates the carotid baroreflex, an effect of which may be resulted from the opening of K(ATP) channels mediated by VR(1). PMID- 15127134 TI - [Effects of beta-adrenoceptor activation on metabolism in neonatal rat cardiomyocytes]. AB - The aim of the present study was to investigate the effects of beta-adrenergic receptor (beta-AR) activation on metabolism in cultured neonatal rat cardiomyocytes. The protein synthesis and total protein content of cardiomyocytes were determined by [(3)H]-leucine incorporation and BCA protein content assay. Cardiomyocyte glucose uptake was measured by [(3)H]-2-deoxy-D-glucose uptake analysis. Adenosine monophosphate activated protein kinase (AMPK) phosphorylation was detected by Western blot. The results showed that sustained stimulation with isoproterenol (ISO), a beta-adrenoceptor agonist, had no effect on [(3)H]-leucine incorporation and total protein content in cardiomyocytes. With beta-AR activation by ISO or NE (pretreated with a selective blocker of the alpha(1) adrenoceptor prazosin) for 48 h, both the glucose uptake and AMPK phosphorylation increased significantly compared with unstimulated cardiomyocytes. These results suggest that although sustained beta-AR activation has no effect on cardiomyocyte protein metabolism, glucose uptake and AMPK activity are increased significantly. The role of these beta-AR activation-induced changes in cardiac hypertrophy remains to be further investigated. PMID- 15127135 TI - ClC-3 expression in the cell cycle of nasopharyngeal carcinoma cells. AB - The immunofluorescence approach, the confocal microscopy and the patch-clamp technique were used to investigate the expression of ClC-3 (one of the candidates of volume-activated chloride channels) and its relationships with the volume activated chloride current and the capacity of regulatory volume decrease (RVD) in the cell cycle of nasopharyngeal carcinoma cells (CNE-2Z cells). The results indicated that CNE-2Z cells expressed ClC-3. ClC-3 was located predominantly inside the cells but not in the membrane. Both the expression level and the distribution of ClC-3 were cell cycle dependent. ClC-3 expression was low in G1 but high in S phase. The cells in G2/M phase possessed an intermediate level of the expression. ClC-3 expression level was negatively correlated to the RVD capacity and amplitude of the volume-activated chloride current in the cell cycle. The results suggest that ClC-3 may be an important factor in the regulation of cell cycle progression, but that it is probably not the chloride channel associated with RVD in these cancer cells. PMID- 15127136 TI - [Protective effect of nitric oxide against hydrogen peroxide-induced hearing loss]. AB - Previous research showed that reactive oxygen species (ROS) play an important role in ototoxity. The present research was to investigate whether nitric oxide, an important neurotransmitter in the inner ear, could prevent hydrogen peroxide induced hearing loss through the nitric oxide/cyclic GMP pathway in guinea pig cochlea. Fifty adult pigmented guinea pigs (250~350 g) of either sex with positive prier reflex were randomly divided into five groups. All of the animals underwent whole cochlear perfusion for two hours. The solution that was perfused into the cochlear of different group was artificial perilymph (AP) for group 1200 micromol/L H2O2 for group 2100 micromol/L L-Arg for group 3, H2O2+L-Arg for group 4 and H2O2+L-Arg+L-NNA for group 5 respectively. Compound action potential (CAP, evoked by click) and cochlear microphonic (CM, evoked by tone burst) were recorded every thirty minutes to show the effects of different reagents on cochlear function. In order to assess cell viability after perfusion, the fluorescent dyes Hoechst that stains all cell nuclei and propidium iodide (PI) that specifically stains nuclei of dead cells, were used. The CAP threshold shifts and CM amplitude decreased after perfusion with H2O2+L-Arg. They were significantly lower than those of H2O2 group. No obvious cell death was noticed after H2O2+L-Arg perfusion, while only 54% of hair cells were alive after H2O2 perfusion. There were no significant differences between the group of H2O2 and that of H2O2+L-Arg+L-NNA group. Our results suggest that nitric oxide may partly be able to protect guinea pigs from hydrogen peroxide-induced hearing loss. PMID- 15127138 TI - Electrophysiological effects of capsaicin on spontaneous activity of rabbit atrioventricular node cells. AB - To study the electrophysiological effects of capsaicin on spontaneous activity of rabbit atrioventricular (AV) node cells, parameters of action potential in AV node were recorded using intracellular microelectrode technique. Capsaicin (1-30 micromol/L) not only decreased the amplitude of action potential, maximal rate of depolarization (V(max)), velocity of diastolic (phase 4) depolarization, and rate of pacemaker firing, but also prolonged the duration of 90% repolarization of action potential (APD(90)) in a concentration-dependent manner. Both application of L-type Ca(2+) channel agonist Bay K8644 (0.5 micromol/L) and elevation of calcium concentration (5 mmol/L) in superfusate antagonized the effects of capsaicin on pacemaker cells. Pretreatment with ruthenium red (10 micromol/L), a capsaicin receptor blocker, did not affect the effects of capsaicin on AV node cells. Capsaicin exerted an inhibitory action on spontaneous activity of AV node cells in rabbits. These effects were likely due to reduction in calcium influx, but were not mediated by VR1. PMID- 15127137 TI - Effects of low-dose capsaicin on L-type calcium current in guinea pig ventricular myocytes. AB - The purpose of this study was to examine the effects of low-dose capsaicin (CAP) on L-type calcium current (I(Ca-L) ) in guinea pig ventricular myocytes and the underlying mechanism. I(Ca-L) was examined in isolated single guinea pig ventricular myocytes by using whole-cell patch clamp technique. CAP (1-25 nmol/L) increased the voltage-dependently activated peak amplitude of I(Ca-L) and downshifted the current-voltage (I-V) curve. CAP (1, 10, 25 nmol/L) increased the peak amplitude of I(Ca-L) from -9.67+/-0.7 pA/pF to -10.21+/-0.8 pA/pF (P>0.05), to -11.37+/-0.8 pA/pF and to -12.84+/-0.9 pA/pF (P<0.05), respectively. CAP 25 nmol/L shifted the steady-state activation curve of I(Ca-L) to the left and changed half activation potential (V(0.5)) from (-20.76+/-2.0) mV to (-26.71+/ 3.0) mV (P<0.05), indicating that low-dose CAP may modify the voltage-dependent activation of calcium channel. Low-dose of CAP did not affect the steady-state inactivation curve of I(Ca-L) or half-recovery time of Ca(2+) channel from inactivation. Ruthenium red (RR, 10 micromol/L), a vanilloid receptor (VR1) blocker, antagonized the effects of low-dose CAP. These results suggest that low dose CAP increases I(Ca-L) mainly by shifting its steady-state activation curve to the left. Such effects may be mediated by VR1. PMID- 15127139 TI - [3T3-L1 adipocytes reduces Kir6.2 channel expression in MIN6 insulin-secreting cells in vitro]. AB - Dysfunction of the pancreatic beta-cell is an important defect in the pathophysiological changes of type 2 diabetes, and type 2 diabetes is evidently associated with obesity. But the role of the adipocyte in the dysfunction of the pancreatic beta-cell remains unknown. In the present study, we examined the direct effects of 3T3-L1 adipocytes on the expression of ATP-sensitive potassium channels (K(ATP) channels) in MIN6 insulin-secreting cells. MIN6 cells were divided into two groups as control group, where MIN6 cells were cultured in normal culture medium, and coculture group, where MIN6 cells were cocultured with differentiated 3T3-L1 adipocytes for 1 week. Semi-quantitative RT-PCR was employed to measure the expression of K(ATP) channel subunit Kir6.2 in MIN6 cells. Fura-2 was used to reflect changes in intracellular calcium concentration ([Ca(2+)](i)) in MIN6 cells. The secretary function of MIN6 cells from both groups was estimated by radioimmunoassay method. The results showed that the Kir6.2 cDNA levels corrected by GAPDH cDNA levels after densitometric analysis were 0.989+/-0.035 in control group and 0.726+/-0.087 in coculture group. The expression of Kir6.2 was significantly decreased in MIN6 cells in the coculture group as compared with that in control. MIN6 cells cocultured with 3T3-L1 adipocytes lost the ability to increase [Ca(2+)](i) when stimulated by tolbutamide (0.1 mmol/L), a highly selective KATP channel closer. In contrast, MIN6 cells in control group had typical responses to tolbutamide with a significant increase in [Ca(2+)](i). The magnitudes to basal levels of [Ca(2+)](i) after tolbutamide stimulation were 1.520+/-0.203 in control and 1.114+/-0.097 in coculture group (P<0.05, n=6). MIN6 cells in control showed a significant increase in insulin secretion from 0.38+/-0.099 mU/min to 2.87+/ 0.248 mU/min after being stimulated by tolbutamide, whereas MIN6 cells in coculture group did not increase insulin secretion when stimulated by tolbutamide (0.21+/-0.055 mU/min to 0.22+/-0.082 mU/min). It is demonstrated that 3T3-L1 adipocytes decrease the expression of K(ATP) channels in MIN6 cells through secreting certain factors, which impair the secretary function of MIN6 cells. The present results indicate that adipocytes are directly involved in pancreatic beta cell dysfunction, which may facilitate the development of type 2 diabetes. PMID- 15127140 TI - Predominant Th2-type response during normal pregnancy of rats. AB - The immunological parameters were analyzed during pregnancy of Lewis rats by the methods of flow cytometry, thymidine incorporation and enzyme-linked immunospot (ELISPOT). MHC II of spleen mononuclear cells (MNCs) and CD11c of periphery blood MNCs was apparently downregulated in late pregnancy, while the costimulatory molecules B7-1 and B7-2 showed no difference. Increased expression of Th2 cytokines (IL-10, IL-4) and TGFbeta was detected in the spleen and peripheral blood MNCs in the third trimester by flow cytometry. No suppression of Th1 cytokine represented by IFNgamma was found. Furthermore, antigen specific proliferation of spleen and peripheral blood MNCs was unchanged, but higher proliferation of MNCs from mesenteric lymph nodes was shown in late pregnancy. There was an inhibition of antigen specific antibody production in pregnancy examined by ELISPOT. These data indicate the immunomodulatory effects of sex hormones in pregnancy, which may be related to the remission of T cell-mediated autoimmune diseases during pregnancy. PMID- 15127141 TI - [Effect of repeated acute hypoxic treatment on the expression of alpha-synuclein in the mouse brain cortex]. AB - An anti-alpha-synuclein (alpha-SYN) monoclonal antibody produced in our laboratory was used to investigate the effect of repeated acute hypoxic treatments on the expression of alpha-SYN in the mouse cerebral cortex. Western blot analysis showed that the expression levels of alpha-SYN in the cortex changed accordingly upon hypoxic exposure times, as that the alpha-synuclein level significantly increased after the first hypoxic exposure and then dropped down to the background level after the fourth hypoxic exposure. Immunohistochemical staining revealed that the alpha-SYN-immunopositive substance was localized not only in the nerve endings, but also within the nuclei of some neurons. The cell density of the neurons with alpha-SYN immunopositive nuclei was increased significantly after the first hypoxic exposure but returned back to control levels after the fourth hypoxic exposure. Our results indicate that both of the alpha-SYN expression level in the brain and the number of the neurons with alpha-SYN positive nuclei are affected by the repeated acute hypoxic treatments and that this modification is hypoxic time-dependent. The mechanism and the physiological significance underlying these changes need to be further investigated. PMID- 15127142 TI - [The causal relationship between dissociation and trauma. A critical review]. AB - This article discusses the accepted view that trauma causes dissociation. We critically evaluate studies that explored this relationship and discuss their shortcomings. In addition, individual differences related to dissociative experiences are discussed. In doing so, we focus on their potential to create pseudo-correlations between dissociative symptoms and self-reports of childhood trauma. After concluding that the causal link between trauma and dissociation is far from self-evident or empirically substantiated, we suggest some avenues for further research. PMID- 15127143 TI - [Heat stroke with alpha coma. A case report]. AB - We report the case of a near-fatal heat stroke in a 41-year-old patient. The comatose patient had a body core temperature of 41.5 degrees C. The clinical course was complicated by systemic inflammatory response syndrome and multiorgan failure. The EEG showed an alpha coma that did not react to external stimuli and, in general, has a poor prognosis. The patient regained consciousness and was discharged from our intensive care unit after 16 days. In the further course cerebral toxoplasmosis developed which was treated with a combination therapy of sulfadiazine and pyrimethamine. The patient was transferred to a neurorehabilitation clinic with a moderate neurological deficit 65 days after heat stroke onset. PMID- 15127146 TI - [Pronounced bilateral mesocochlear hearing impairment from 40-45 dB in spite of regular TEOAE, DPOAE and inconspicuous click-BERA]. AB - An objective screening method for assessment of inner ear function of babies and small children can be carried out by measuring TEOAE and DPOAE. In particular, TEOAE can be used as a cheap and quick method to determine, with very high sensitivity and moderate specificity, whether middle to severe peripheral hearing impairment is present. In order to assess the threshold more precisely, the click BERA is routinely used. We present a case report of a 6 year old girl with regular TEOAE and DPOAE, and who also had bilateral normal thresholds in the click-BERA. With subjective audiometry, the girl could be diagnosed having bilateral mesocochlear hearing impairment (so called cookie-bite audiogram) ranging at 40-45 dB. The child was helped considerably by fitting of hearing aids bilaterally. In order to determine the auditory ability of children and to provide adequate care, subjective as well as objective tests should be used. PMID- 15127147 TI - [Choristoma in the vocal fold]. AB - Choristoma in the larynx is rare and related to thyroid or glial tissue. The manifestation of salivary gland tissue in the larynx has not been reported to date. We present the case of an 80 year old male complaining of hoarseness and productive coughing. A left side tumourous swelling was seen in the larynx with intact vocal cord mobility. Using microlaryngoscopy the mass was resected without any intra- or postoperative problems. Histologically, a choristoma-heterotopic salivary gland tissue in the muscle and fat tissue--was found. The mucosa was intact and there were no signs of malignancy. Differential diagnosis of such masses in the larynx include benign lesions as well as specific infections, e.g. tuberculosis, sarcoidosis, amyloidosis and Wegener's granulomatosis. Especially in non-smokers, sarcoma, lymphoma and melanoma should be separated from the frequent squamous cell carcinoma of the laryngeal tissue. In rare cases, heterotopic tissue can mimic a tumourous mass. Intralaryngeal resection is the therapy of choice and should be recommended to the patient. PMID- 15127148 TI - [Pulse-synchronous tinnitus]. PMID- 15127144 TI - [Therapy resistance to antidepressants. Definition, prevalence, predictors, and interventional possibilities]. AB - A major problem in the medical treatment of depression is that not all patients respond sufficiently to drug treatment. This applies particularly when the therapeutic aim is complete relief from symptoms, not merely response. This article presents an overview of the definitions and epidemiologic, predictive, and therapeutic aspects of the problem. PMID- 15127149 TI - [Significance of voice constitution as a predisposition for occupational voice disorders]. AB - Occupational voice disorders have been increasing for years. The aim of this study was to examine whether a constitutionally weak voice should be regarded as a risk factor for developing such voice disorders. In a prospective study, 15 female teacher students with a normal vocal constitution were compared with 18 students with constitutionally weak voices during teaching practice. There was a significant difference in the mean fundamental frequency of both groups after teaching for 30 min. Students with constitutionally weak voices tended to have increased values. Taking into consideration the physiological aspects of an increasing fundamental frequency as a sign of vocal fatigue, a constitutionally weak voice has to be regarded as a potential risk factor for developing a voice disorder in vocally intensive occupations. To prevent future vocal problems by appropriate prophylactic intervention (i.e. voice therapy, introduction to vocal hygiene), a phoniatric examination of vocal constitution and endurance is recommended at the beginning of a voice dependent professional career. PMID- 15127150 TI - [Selected emergencies in operative dermatology]. AB - The large number of surgical procedures in many subspecialty areas of dermatology, such as dermatologic oncology, phlebology and aesthetic dermatology, raises the question of possible complications and emergency situations. Along with common complications such as post-operative bleeding, infections or flap necrosis, there are emergency situations which require immediate intervention to avoid significant harm to the patient. One must distinguish between dermatologic situations which require emergency surgery and emergencies that arise during or after dermatological surgery. PMID- 15127151 TI - [Developments in chromate allergy in the German construction industry]. AB - For decades potassium dichromate has been the most important allergen in the construction industry. In Scandinavian countries the prevalence of potassium dichromate sensitization declined following the introduction of low-chromate cement. In contrast, analysis of our register in Northern Bavaria and the data of workers compensation board show no significant decline in potassium dichromate sensitization in the German construction industry during the 1990s. In 1993, German legislation provided an Approved Code of Practice and the cement manufacturers committed themselves to reduce the level of water-soluble chromates to less than 2 ppm in bag cement. Since 2000 this agreement has also included ready-mixed concrete. It remains to be seen, if sensitization against potassium dichromate will decline in the upcoming years. Based on our data, it would be desirable if only low-chromate cement was used in the construction industry in Germany as in Scandinavia. PMID- 15127153 TI - [The "Wurzburg T". A concept for optimization of early multiple trauma care in the emergency department]. AB - INTRODUCTION: Anaesthesia management, radiological diagnostic and the concept of damage control surgery should be combined in the resuscitation room. Defined clinical targets and their realisation are a CT-scan and complete damage control surgery in the shock room. Furthermore minimised patient transfer and positioning with continuous access to the head, upper parts of the body and anaesthesia machine should be realised during diagnostic procedures. METHODS: Based on a carbon-slide fixed on a turntable and innovative alignment of diagnostic devices, a three phase treatment algorithm has been established. RESULTS: Phase A includes primary survey, anaesthetic management and ultrasound examination. Following a turn of the table conventional x-ray diagnostic is assessed in phase B. Tracks for the slide enable immediate transfer to a spiral CT-scan without additional patient positioning (phase C). Following complete CT-scan rearrangement of the table to phase A facilitates immediate damage control surgery. To accelerate device operation and treatment the integrated anaesthesia workstation is ceiling mounted and manoeuvres close to the patient. CONCLUSIONS: This concept realizes complete diagnostic procedures and damage control surgery without time consuming patient transfer or rearrangement. PMID- 15127152 TI - [Regulation of spinal nociceptin expression by neuropathic pain]. AB - Nociceptin is the endogenous ligand of a new opioid receptor, the opioid receptor like-1 (ORL1) receptor. Chronic inflammatory pain causes an increase in the expression of nociceptin and the ORL1 receptor in the dorsal horn of rat spinal cord, thus indicating an involvement of the endogenous nociceptin/ORL1 system in mechanisms of pathological pain. This study investigates the influence of neuropathic pain on the expression of nociceptin using immunohistochemistry. To induce neuropathic pain, a ligation of the sciatic nerve was performed in 12 rats under general anesthesia. A sham operation was performed in 12 rats of the control group. Nerve ligation caused a significant ipsilateral thermal hyperalgesia, a typical sign of neuropathic pain. The paw withdrawal latency was decreased by 45.7+/-4.9% ( p<0.05) at day 5 and by 37.3+/-1.8% ( p<0.05) at day 10. Although hyperalgesia was fully present after 5 days, no changes in nociceptin immunoreactivity in the lumbar spinal cord were detected at this time point. Ten days after nerve ligation, there was a 2.46+/-0.38 fold ( p<0.05) bilateral increase in nociceptin immunoreactivity in the lamina superficiales (I and II), with a notable increase in the inner lamina II at the level of L4. Further investigations are necessary to elucidate the relationship between neuropathic pain, the nociceptin-ORL1 receptor system and potential therapeutic options. PMID- 15127154 TI - [Legal questions in radio-oncology. Demands on patient information and documentation of the therapy with examples of current legal cases concerning specialization in radiotherapy/radio-oncology]. AB - BACKGROUND: In recent years, also due to the media, the medical information a physician gives to his patients, the methods of medical treatment and their consequences have increasingly come to the attention of the general public. As a result, to avoid claims for compensation because of faulty treatment or even to evade prosecution, the demands on medical information given before or during treatment are getting greater and greater. Generally, claims for compensation are not based on the accusation of wrong treatment, because this accusation is difficult to prove. As a rule-as a way out of difficulties-these claims for compensation are based on the accusation of mistaken or incomplete information, because if one cannot prove any wrong treatment, faulty information can (nearly) always be proven. METHODS: Methodically, showing the principles that must be followed when giving information on radiotherapy and when documenting it, also taking examples of current legal cases into account. Short survey of the question of liability and the future importance of principles for medical treatment. CONCLUSION: Every attending physician should feel obliged to pay attention to legal questions concerning medical subjects, though judgments on the contents and the extent of the information that must be given to patients are complex and difficult to understand for anybody not experienced in law. Only so the ever increasing tendency of people to try to make financial profit on information that they consider mistaken or incomplete can be opposed and a lawsuit can be won, if it cannot be avoided. PMID- 15127155 TI - Linac radiosurgery versus whole brain radiotherapy for brain metastases. A survival comparison based on the RTOG recursive partitioning analysis. AB - BACKGROUND AND PURPOSE: For patients with inoperable brain metastases, whole brain radiotherapy (WBRT) has been the standard treatment for decades. Radiosurgery is an effective alternative strategy, but has failed to show a substantial survival benefit so far. The prognostic factors derived from the RTOG recursive partitioning analysis (RPA) provide a framework that allows a nonrandomized comparison of the two modalities. PATIENTS AND METHODS: From 1991 to 1998, 117 patients with one to three previously untreated cerebral metastases underwent single-dose linac radiosurgery (median dose 20 Gy) without adjuvant WBRT. After radiosurgery, 26/117 patients (22%) had salvage WBRT, radiosurgery or neurosurgical resection of recurrent (4/117) and/or new (24/117) metastases. Survival of these patients was compared to a historical group of 138 patients with one to three lesions treated by WBRT (30-36 Gy/3-Gy fractions) from 1978 to 1991; only nine of these patients (7%) had salvage WBRT. All patients were classified into the three RPA prognostic classes based on age, performance score, and presence of extracranial tumor manifestations. RESULTS: In RPA class I (Karnofsky performance score > or = 70, primary tumor controlled, no other metastases, age < 65 years), radiosurgery resulted in a median survival of 25.4 months (n = 23, confidence interval [CI] 5.8-45.0) which was significantly longer than for WBRT (n = 9, 4.7 months, CI 3.8-5.5; p < 0.0001). In RPA class III (Karnofsky performance score < 70), no significant difference in survival between radiosurgery (n = 20, 4.2 months, CI 3.2-5.3) and WBRT (n = 68, 2.5 months, CI 2.2-2.8) was found. In RPA class II (all other patients), radiosurgery produced a small, but significant survival advantage (radiosurgery: n = 74, 5.9 months, CI 3.2-8.5, WBRT: n = 61, 4.1 months, CI 3.4-4.9; p < 0.04). CONCLUSION: Radiosurgery in patients with one to three cerebral metastases results in a substantial survival benefit only in younger patients with a low systemic tumor burden when compared to WBRT alone. It cannot be excluded that this effect is partially caused by the available salvage options after radiosurgery. PMID- 15127156 TI - Prognostic factors for brain metastases after whole brain radiotherapy. Data from a single institution. AB - PURPOSE: Prognostic factors for overall survival of patients treated for brain metastases with whole brain radiotherapy (WBRT) at a single institution were retrospectively evaluated, and the validity of the RTOG recursive partitioning analysis (RPA) for prognostic classes was assessed. PATIENTS AND METHODS: The data of all patients (n = 268) with brain metastases from solid tumors homogeneously treated between 01/1997 and 09/1999 at the University of Heidelberg, Germany, with WBRT without surgery or radiosurgery were reviewed. 13 different patient- and therapy-related variables were evaluated for prognosis. Second, a grouping of the study cohort was performed according to the RTOG RPA prognostic classes. RESULTS: Median survival of the whole population after the start of WBRT was 3.8 months. The 1-year survival rate was 19%. Multivariate analysis revealed that only the Karnofsky performance status, control of the primary and no extracranial disease were independent prognostic factors for overall survival. These are also the main determinants of the RTOG RPA classes. Applying the RTOG RPA classes to the authors' data set revealed three subgroups with significantly different prognosis. CONCLUSION: Based on this analysis, prognostic factors for survival after WBRT in patients with brain metastases could be identified. A total of 19% (n = 44/232) survived > or = 1 year, whereas overall survival was poor. The potential value of the RPA classes in estimating the patient's prognosis could be confirmed. PMID- 15127157 TI - [CT-Guided brachytherapy. A novel percutaneous technique for interstitial ablation of liver metastases]. AB - PURPOSE: To assess safety and efficacy of CT-guided brachytherapy of liver malignancies. PATIENTS AND METHODS: 21 patients with 21 liver malignancies (19 metastases, two primary liver tumors) were treated with interstitial CT-guided brachytherapy applying a (192)Ir source. In all patients, the use of image-guided thermal tumor ablation such as by radiofrequency or laser-induced thermotherapy (LITT) was impeded either by tumor size > or = 5 cm in seven, adjacent portal or hepatic vein in ten, or adjacent bile duct bifurcation in four patients. Dosimetry was performed using three-dimensional CT data sets acquired after CT guided positioning of the brachytherapy catheters. RESULTS: The mean tumor diameter was 4.6 cm (2.5-11 cm). The mean minimal tumor dose inside the tumor margin amounted to 17 Gy (12-20 Gy). The proportion of the liver parenchyma exposed to > 5 Gy was 18% (5-39%) of total liver parenchyma minus tumor volume. Nausea and vomiting were observed in six patients after brachytherapy (28%). One patient demonstrated obstructive jaundice due to tumor edema after irradiation of a metastasis adjacent to the bile duct bifurcation. We commonly encountered asymptomatic increases of liver enzymes. Local control rates after 6 and 12 months were 87% and 70%, respectively. CONCLUSION: CT-guided brachytherapy is safe and effective. This technique displays broader indications compared to image guided thermal ablation by radiofrequency or LITT with respect to tumor size or localization. PMID- 15127158 TI - [Influence of preoperative (hyperthermic) radiochemotherapy on manometric anal sphincter function in locally advanced rectal cancer]. AB - BACKGROUND AND PURPOSE: Preoperative radiochemotherapy (RCT) followed by curative surgery is a well-accepted therapeutic option in the treatment of advanced rectal cancer. Usually, the anal sphincter is located in the irradiation area of a preoperative RCT regime. The aim of this study is to evaluate the influence of preoperative RCT on anal sphincter function. PATIENTS AND METHODS: Between 1994 and 2000, 102 patients with rectal cancer stage uT3/uT4 were analyzed. All patients underwent radiotherapy with 45 Gy (5 x 1.8 Gy) including two cycles of 5 fluorouracil (5-FU)/leucovorin (folinic acid) chemotherapy. 46 patients were treated additionally with up to five sessions of locoregional hyperthermia. The sphincter function was analyzed by perfusion manometry before preoperative therapy and 4 weeks after pretreatment had been finished. For statistics, the Wilcoxon signed rank test and Mann-Whitney U-test were used (SPSS 9.0 for Windows((R))). RESULTS: The mean value of all 102 patients showed a significant reduction of the mean maximum resting pressure from 97 to 89 mmHg (p = 0.02). For the mean maximal squeeze pressure no significant difference could be shown (178 vs. 176 mmHg). For patients with distal (900 ml/m(2) was reached. Repeated haemodynamic measurements were done at baseline (t(b)), at the end point (t(ep)) then at 30 min and 60 min after the end point was reached (t(30), t(60)). Cardiac output, stroke volume, extravascular lung water (EVLW), and oxygen delivery was determined at each assessment point. For statistical analysis two-way ANOVA was used. MEASUREMENTS AND RESULTS: ITBVI, cardiac index, and oxygen delivery index increased significantly at t(ep) and remained elevated for t(30) and t(60), but there was no significant difference between the two groups. The increase in the ITBVI by 100 ml of infusion was similar in both groups (HES: 26+/-19 ml/m(2) vs GEL: 30+/-19 ml/m(2)). EVLW, remained unchanged, and there was no significant difference between the groups (HES, t(b): 8+/-6, t(60): 8+/-6; GEL, t(b): 8+/-3, t(60): 8+/-3 ml/kg). The PaO(2)/FiO(2) did not change significantly over time or between groups (HES, t(b): 207+/-114, t(60): 189+/-78; GEL, t(b): 182+/-85, t(60): 182+/-85 mmHg). CONCLUSION: The results of this study indicate that both HES and GEL infusions caused similar short-term change in ITBVI in septic shock, without increasing EVLW or worsening oxygenation. PMID- 15127187 TI - Relapse of SARS upon tapering corticosteroid. PMID- 15127188 TI - The best place to commit the perfect murder. PMID- 15127189 TI - Consciousness monitoring in ventilated patients: bispectral EEG monitors arousal not delirium. AB - OBJECTIVE: Bispectral index (BIS) is being evaluated as a monitor of consciousness, yet it is unclear what components of consciousness (i.e., arousal vs. content of consciousness) the BIS measures. This study compared BIS levels to well-validated clinical measures of arousal and the presence or absence of delirium. DESIGN: A prospective, blinded, observational cohort study. PATIENTS: 124 mechanically ventilated, adult, medical ICU patients. MEASUREMENTS AND RESULTS: Using BIS 3.4 and BIS-XP 4.0 algorithms, BIS values were calculated immediately prior to clinical assessments. The clinical assessments included the Richmond Agitation-Sedation Scale (RASS) and presence or absence of delirium using the Confusion Assessment Method for the ICU. A total of 484 assessments were collected among 124 patients. BIS-XP values demonstrated greater correlation with RASS than BIS 3.4 ( R(2)=0.36 vs. 0.20), although considerable overlap of BIS-XP scores remained across RASS levels. Median BIS-XP values for delirious and nondelirious observations were 74 and 96, respectively, while BIS 3.4 values were 91 and 96, respectively. However, neither BIS 3.4 nor BIS-XP were significantly associated with delirium after controlling for RASS value. CONCLUSIONS: In comparison with clinical measures of arousal in mechanically ventilated patients, BIS-XP algorithm demonstrated stronger correlation with RASS levels than did BIS 3.4, yet marked overlap across different levels of arousal persist using both algorithms. After controlling for level of arousal, neither BIS-XP nor BIS 3.4 algorithms distinguished between the presence and absence of delirium. PMID- 15127190 TI - Transcardiac conductance for continuous measurement of left ventricular volume: validation vs. angiography in patients. AB - OBJECTIVE: To test the feasibility of the transcardiac conductance (TCC) method for continuous, on-line measurement of absolute left ventricular (LV) volume and to validate the method by comparison with biplane angiography. DESIGN AND SETTING: Prospective clinical feasibility and validation study in a cardiac catheterization laboratory in a university hospital. PATIENTS AND INTERVENTIONS: Ten patients scheduled for electrophysiological studies ( n=5), percutaneous transluminal coronary angioplasty ( n=3), and left- and right-sided cardiac catheterization ( n=2) were enrolled in the feasibility study. Twenty patients scheduled for diagnostic left- and right-sided cardiac catheterization were included in the validation study. The latter were studied at baseline and during right atrial pacing 30 beats/min above baseline. MEASUREMENTS AND RESULTS: In the feasibility study satisfactory ventricular volume signals were obtained by TCC in eight of ten patients. In the validation study calibration factors (alpha and V(p)) for TCC were obtained by thermodilution and hypertonic saline dilution, to yield absolute LV volume. Results indicate a good linear correlation with angiographic volume ( R(2)=0.78) with an intercept of 10+/-15 ml, not significantly different from 0 and slope of 1.17+/-0.16. Mean calibration factors alpha and V(p) were 0.017+/-0.002 (interpatient variability 0.018) and 75.1+/-0.4 ml (interpatient variability 35.4 ml), respectively. CONCLUSIONS: The TCC method provides on-line and continuous LV volume signals in patients in a relatively noninvasive way. Calibration yields absolute LV volumes with a good linear correlation in comparison to biplane LV angiography. TCC appears to be a promising methodology for monitoring absolute LV volume in the ICU. PMID- 15127191 TI - Increased incidence and severity of the systemic inflammatory response syndrome in patients deficient in mannose-binding lectin. AB - OBJECTIVE: To determine whether pediatric PICU patients with mannose-binding lectin (MBL) gene polymorphisms associated with low levels of the functional protein have an increased risk of developing sepsis and SIRS. DESIGN AND SETTING: A prospective, observational cohort study in a 22-bed PICU in a tertiary referral centre. PATIENTS: One hundred consecutive admissions to a PICU with at least one organ system failure longer than 12 h. Patients were classified into those with infectious or non-infectious insults as the primary reason for intensive care admission. Patients were followed to determine which developed sepsis or non infection related SIRS using standard criteria. MEASUREMENTS AND RESULTS: Of the 100 patients 50 had infectious and 50 had non-infectious insults as the precipitant for admission. 42 patients had variant MBL alleles (determined by MBL 2 gene exon 1 and promoter polymorphisms) and were significantly over-represented amongst the 59 patients that developed SIRS. This effect was not explained by differences in age, sex or ethnicity and was seen in both the infection and non infection subgroups. In patients with infection, variant MBL alleles were associated with increased systemic response (2/15 with localised infection, 10/19 with sepsis and 12/16 with septic shock). MBL serum levels showed close concordance with the genotype and indicated that MBL levels less than 1000 ng/ml are associated with a greatly increased risk of SIRS. CONCLUSIONS: MBL-2 exon 1 polymorphisms with low serum levels of functional MBL protein are associated with a greatly increased risk of developing SIRS and of progression from infection to sepsis and septic shock in paediatric ICU patients. PMID- 15127193 TI - Standardization of intravenous insulin therapy improves the efficiency and safety of blood glucose control in critically ill adults. AB - OBJECTIVE: Aggressive glycemic control improves mortality and morbidity in critically ill adults, however implementation of such a strategy can be logistically difficult. This study evaluates the efficiency and safety of a nurse managed insulin protocol in critically ill adults. DESIGN: Combined retrospective prospective before-after cohort study. SETTING: Twenty-one bed, medical/surgical ICU in a tertiary care hospital. PATIENTS: Two cohorts of 50 consecutive ICU patients requiring insulin infusions. INTERVENTION: Patients in the control cohort received insulin infusions titrated according to target blood glucose ranges and sliding scales at the physician's discretion. Patients in the interventional cohort received an insulin infusion adjusted using a standardized protocol targeting a blood glucose of 4.5-6.1 mmol/l (81-110 mg/dl). MEASUREMENTS AND MAIN RESULTS: Efficiency was measured by comparing the time to reach, and the time spent within, the target range between cohorts. Safety was assessed by comparing the incidence of severe hypoglycemia, the frequency of rescue dextrose administration and the cumulative time that the infusion was held for hypoglycemia between cohorts. Patients in the interventional cohort reached their target more rapidly (11.3+/-7.9 vs 16.4+/-12.6 h; p=0.028) and maintained their blood glucose within the target range longer (11.5+/-3.7 vs 7.1+/-5.0 h/day; p<0.001) than controls. The standardized protocol yielded a four-fold reduction in the incidence of severe hypoglycemia (4 vs 16%; p=0.046) and reduced the median frequency of dextrose rescue therapy (0 [0-0.91] vs 0.17 [0-1.2] episodes/patient per day; p=0.01) as compared to controls. CONCLUSION: Standardization of intensive insulin therapy improves the efficiency and safety of glycemic control in critically ill adults. PMID- 15127192 TI - Lipopolysaccharide-binding protein in critically ill neonates and children with suspected infection: comparison with procalcitonin, interleukin-6, and C-reactive protein. AB - OBJECTIVE: To evaluate markers of infection in critically ill neonates and children, comparing lipopolysaccharide-binding protein (LBP) with procalcitonin (PCT), interleukin-6 (IL-6), and C-reactive protein (CRP). DESIGN AND SETTING: Prospective, observational study in the level III multidisciplinary neonatal and pediatric intensive care unit. PATIENTS: Sixty patients with systemic inflammatory response syndrome (SIRS) and suspected infection classified into two groups: SIRS/sepsis ( n=33) and SIRS/no sepsis ( n=27). We included 29 neonates aged less than 48 h (neonates <48 h), 12 neonates older than 48 h (neonates >48 h), and 19 children. Median disease severity was high in neonates aged under 48 h and moderate in neonates aged over 48 h and children. INTERVENTIONS: Serum LBP, PCT, IL-6, and CRP were measured on two consecutive days. Area under the receiver operating characteristic (ROC) curve (AUC), sensitivity, specificity, and predictive values were evaluated. RESULTS: Serum LBP was higher in patients with SIRS/sepsis than in patients with SIRS/no sepsis. AUC for LBP on the first day of suspected infection was 0.89 in the younger neonates, 0.93 in the older neonates, and 0.91 in children. CONCLUSIONS: In critically ill neonates aged under 48 h LBP on the first day of suspected infection is a better marker of sepsis than IL-6 and PCT, and is similar to CRP. In critically ill neonates aged over 48 h and children LBP is a better marker than IL-6 and CRP, and is similar to PCT. PMID- 15127194 TI - Body temperature alterations in the critically ill. AB - OBJECTIVE: To determine the incidence of body temperature (BT) alterations in critically ill patients, and their relationship with infection and outcome. DESIGN: Prospective, observational study. SETTING. Thirty-one bed, medico surgical department of intensive care. PATIENTS: Adult patients admitted consecutively to the ICU for at least 24 h, during 6 summer months. INTERVENTIONS: None. RESULTS: Fever (BT > or =38.3 degrees C) occurred in 139 (28.2%) patients and hypothermia (BT< or =36 degrees C) in 45 (9.1%) patients, at some time during the ICU stay. Fever was present in 52 of 100 (52.0%) infected patients without septic shock, and in 24 of 38 (63.2%) patients with septic shock. Hypothermia occurred in 5 of 100 (5.0%) infected patients without septic shock and in 5 of 38 (13.1%) patients with septic shock. Patients with hypothermia and fever had higher Sequential Organ Failure Assessment (SOFA) scores on admission (6.3+/-3.7 and 6.4+/-3.3 vs 4.6+/-3.2; p<0.01), maximum SOFA scores during ICU stay (7.6+/-5.2 and 8.2+/-4.7 vs 5.4+/-3.8; p<0.01) and mortality rates (33.3 and 35.3% vs 10.3%; p<0.01). The length of stay (LOS) was longer in febrile patients than in hypothermic and normothermic (36 degrees C> expiratory length, P<0.05) to patients with a tumor in the lower lung region (inspiratory length = expiratory length). dMRI is a simple, non invasive method to evaluate diaphragmatic motion and shortening in volunteers and patients during the breathing cycle. Tumors of the lower lung region have a significant influence on shortening of the diaphragm. PMID- 15127222 TI - Whole-brain vascular reactivity measured by fMRI using hyperventilation and breath-holding tasks: efficacy of 3D prospective acquisition correction (3D-PACE) for head motion. AB - Functional MR imaging (fMRI) study using hyperventilation and breath-holding task has been reported to be one of the non-invasive methods to examine whole-brain vascular reactivity. The purpose of this study was to evaluate the efficacy of a method for 3D prospective detection and correction of head motion (3D-PACE) in a study of whole-brain vascular reactivity using hyperventilation and breath holding tasks. Eight healthy volunteers were scanned using an fMRI protocol of hyperventilation and breath-holding task blocks at 3 T in separate runs with and without 3D-PACE. In two subjects, two more runs with and without 3D-PACE were repeated. The mean total number of activated voxels +/- standard deviation was 26,405.3+/-1,822.2 in the run with 3D-PACE and 17,329.9+/-2,766.3 in the run without 3D-PACE ( P<0.05), although there is some intersubject variation regarding the effect of 3D-PACE. In the two subjects whose performed two more runs, the number of activated voxels were smaller in the run without 3D-PACE than even in the run with 3D-PACE performed later. We conclude that 3D-PACE is beneficial for fMRI studies of whole-brain vascular reactivity induced by hyperventilation and breath-holding. PMID- 15127224 TI - Quantitative determination of mosaic GFP gene expression in tobacco. AB - A specific form of gene silencing that was observed visually as a mosaic distribution of fluorescent and non-fluorescent cells apparently dispersed at random within tissues was found in a few green fluorescent protein (GFP) transformed tobacco lines. To characterize this event quantitatively, we studied flow cytometric measurements in GFP-expressing and -silenced cells in T1 and T2 progeny of four selected plants. The proportion of silenced cells varied considerably among the T1 lines but with notable genotype differences. Mosaic expression was inherited into the T2 generation in which the majority of progenies tested exhibited a level of silencing similar to that of their T1 parental plants. However, in some T2 progenies segregation, evident as a decrease or increase in the proportion of fluorescent cells, was observed. We discuss several factors, such as copy number, promoter activity or polyploidy, that may be the possible causes of the gene silencing, but none sufficiently explain the appearance of the mosaic distribution. PMID- 15127223 TI - Heritable transgene expression pattern imposed onto maize ubiquitin promoter by maize adh-1 matrix attachment regions: tissue and developmental specificity in maize transgenic plants. AB - Matrix attachment regions (MARs) have been used to enhance transgene expression and to reduce transgene expression instability in various organisms. In plants, contradictory data question the role of MAR sequences. To assess the use of MAR sequences in maize, we have used two well-characterized MARs from the maize adh-1 region. The MARs have been cloned either 5' to or at both sides of a reporter gene expression cassette to reconstitute a MAR-based domain. Histochemical staining revealed a new transgene expression pattern in roots of regenerated plants and their progeny. Furthermore, MARs systematically induced variegation. We show here that maize adh-1 MARs are able to modify transgene expression patterns as a heritable trait, giving a new and complementary outcome following use of MARs in genetic transformation. PMID- 15127225 TI - Regulation by phosphorylation of Pho81p, a cyclin-dependent kinase inhibitor in Saccharomyces cerevisiae. AB - Cyclins, cyclin-dependent kinases (CDK) and their inhibitors play a critical role in many biological processes. In yeast, the ankyrin repeat protein Pho81p, by being an inhibitor of the Pho85p-Pho80p cyclin-dependent protein kinase complex, transcriptionally regulates the production of repressible acid phosphatase, encoded by the PHO5 gene. Recent studies in our laboratory showed that Pho81p is phosphorylated by the Pho80p-Pho85p CDK complex in vitro; and, to determine the significance of the phosphorylation, we used site-directed mutagenesis to alter the potential phosphorylation sites for this kinase complex. The resulting mutations were introduced into a yeast strain containing a deletion of PHO81 and the effect of the mutation on PHO5 expression was assayed. Results suggest that phosphorylation of particular residues within Pho81p is crucial for its activity as an inhibitor. Studies using a green fluorescent protein-Pho81p fusion and Western analysis indicate that the localization and half-life of the mutants are similar to wild-type Pho81 proteins. However, an in vivo binding assay indicates that the mutant Pho81p is deficient in binding to the Pho80p-Pho85p kinase complex. These findings support the observation that the mutant fails to inhibit kinase activity in low phosphate. These studies provide insight into the mechanism of regulation of CDK inhibitor activity. PMID- 15127226 TI - Saccharomyces cerevisiae translational activator Cbs2p is associated with mitochondrial ribosomes. AB - A characteristic feature of the mitochondrial expression system in Saccharomyces cerevisiae is the requirement for gene-specific translational activator proteins. Translation of mitochondrial apocytochrome b mRNA requires the nucleus-encoded proteins Cbs1p and Cbs2p. These proteins are thought to tether cytochrome b mRNA to the mitochondrial inner membrane via binding to the 5' untranslated mRNA leader. Here, we demonstrate by the use of affinity chromatography and coimmunoprecipitation that Cbs2p interacts with the mitoribosomes. We further provide evidence that the C-terminus of Cbs2p is important for ribosome association, while the N-terminal portion is essential for the formation of homomeric structures. PMID- 15127227 TI - [Mixed epithelial and stromal tumor of the kidney]. AB - We describe a case of mixed epithelial and stromal tumor of the kidney. The tumor was identified in a 63 years old woman and retroperitoneoscopically resected. Immunohistochemistry, electron microscopy and fluorescence in situ hybridization were performed. Grossly, the tumor was well-circumscribed with a diameter of 8 cm and consisted of solid and cystic components. Microscopically, the spindle cell component ranged in appearance from scar-like fibrous tissue to leiomyoma-like fascicles. Epithelial elements were composed of tubules with variable lining. The tubules were focally transformed into large cysts. Estrogen and progesteron receptors were detected in the nuclei of the spindle cells. Electron microscopy confirmed a smooth-muscle differentiation of the spindle cells. Previously, such tumors were published as adult mesoblastic nephroma or cystic hamartoma of the renal pelvis. Its predominance in females with a history of long-term estrogen replacement or the history of long-term sex-steroid use in a male patient, combined with the expression of estrogen und progesteron receptors in the spindle cells suggest that hormonal influence is relevant for the development of these tumors. PMID- 15127228 TI - [Isolated Rosai-Dorfman disease]. AB - A case of extranodal sinus histiocytosis with massive lymphadenopathy (ENSHML; Rosai-Dorfman disease) is reported. The patient presented with a history of intracranial tumour and exophthalmus. Clinical examination found a large mass in the left orbit and paranasal sinuses. Excisional biopsy showed a dense fibrous tissue with an infiltrate rich in macrophages. Further evaluation revealed a retroperitoneal mass with consecutive ureteral stenosis. Further histological and immunohistochemical investigation of the orbital mass, now in suspicion of a systemic disease showed an infiltrate of S-100-positive histiocytes and emperipolesis allowing the diagnosis of extranodal sinus histiocytosis. The correct histologic diagnosis was delayed due to the unusual and isolated extranodal localisation of the disease. The literature concerning extranodal manifestations of Rosai-Dorfman disease is reviewed. We suggest the additional evaluation of such rare and unusual cases in experienced reference centers. PMID- 15127229 TI - Population pharmacokinetics of cisplatin in adult cancer patients. AB - PURPOSE: To characterize the pharmacokinetics of the anticancer agent cisplatin, and explore the influence of patient covariates and interoccasion variability on drug disposition. METHODS: Data were obtained from 285 patients (519 complete curves; 3483 plasma samples) who received the drug as a 3-h intravenous infusion at a mean dose of 144 mg (range 75-210 mg). The population model was built with the use of NONMEM, performing generalized-additive modeling to identify candidate covariates including body-surface area (BSA), age, sex, height, weight, hematocrit, total protein, albumin, serum creatinine, and creatinine clearance, and using a backward deletion protocol to obtain the final models for clearance (CL) and volume of distribution (V). RESULTS: The final model was a one compartment linear model with BSA (in meters squared) as the only significant covariate that impacted on both CL and V: TVCL (in liters per hour)=51.7+26.3x(BSA-1.855) and TVV (in liters)=41.1+24.6x(BSA-1.855), where TVCL and TVV are referred to as typical values that could be used a priori in dosage regimen design. The interindividual and interoccasion variability estimates for CL and V were 16.82 and 20.35%, and 13.93 and 22.91%, respectively. CONCLUSION: A population pharmacokinetic model for cisplatin has been developed that incorporates measures of body size to predict clearance. In this patient population, cisplatin pharmacokinetics were not associated with age, sex, or measures of renal dysfunction. PMID- 15127230 TI - Phase II study of weekly docetaxel combined with cisplatin in patients with advanced non-small-cell lung cancer. AB - PURPOSE: To evaluate the safety and efficacy of the combination of cisplatin on day 1 and docetaxel on days 1, 8 and 15 every 4 weeks for the treatment of previously untreated patients with non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: A group of 38 patients with advanced or metastatic NSCLC who had not received prior treatment and who were aged under 75 years were enrolled. The patients received intravenous infusions of docetaxel (25 mg/m2, days 1, 8, 15) and cisplatin (80 mg/m2, day 1), followed by a week of rest. RESULTS: Six patients had grade 3/4 neutropenia (18%), but there were no episodes of neutropenic fever. Nonhematologic toxicities were also mild. There were 12 partial responses for an objective response rate of 31.6%. The median survival was 11.8 months, and the 1-year survival rate was 46.5%. CONCLUSION: Cisplatin combined with weekly administration of docetaxel is efficacious against NSCLC with low hematotoxicity, and this schedule may be an alternative for the treatment of NSCLC. PMID- 15127231 TI - Weekly administration of topotecan and paclitaxel in pretreated advanced cancer patients: a phase I/II study. AB - PURPOSE: This study was a phase I/II, cohort, dose-escalation trial of topotecan and paclitaxel. Its aim was to determine the dose-limiting toxicity (DLT) of the combination and to define the maximum tolerated dose (MTD), as a recommended dose for phase II, as well as to get preliminary data on the efficacy (activity) of the drug in pretreated patients with ovarian cancer, small-cell lung cancer (SCLC) and non-small-cell lung cancer (NSCLC). METHODS: Included in the study were 52 pretreated patients, 19 with ovarian cancer, 20 with SCLC and 13 with NSCLC. The doses of topotecan were escalated from 1.25 to 2 mg/m2 and of paclitaxel from 60 to 80 mg/m2. A minimum of four patients were included at each of the six levels of dose escalation. RESULTS: We found that DLT due to grade 3 and 4 myelotoxicity was at levels 5 and 6 at doses of 1.75 and 80 mg/m2 (level 5) and 2 and 80 mg/m2 (level 6) for topotecan and paclitaxel, respectively. The MTD and recommended accepted doses are 1.75 mg/m2 for topotecan and 70 mg/m2 for paclitaxel. Of the 52 patients, 17 (33%) showed a response: 1 complete response (1.92%) and 16 partial responses (30.77%). CONCLUSIONS: Topotecan combined with paclitaxel administered once weekly for three consecutive weeks repeated for every 28 days resulted in well-tolerated toxicity at doses of 1.75 and 70 mg/m2, respectively, and a response rate of 33% in pretreated cancer patients. PMID- 15127232 TI - Clinical and pharmacokinetic study of docetaxel in elderly non-small-cell lung cancer patients. AB - PURPOSE: To evaluate the usefulness and pharmacokinetics of docetaxel in the treatment of elderly patients with advanced non-small-cell lung cancer. PATIENTS AND METHODS: Chemotherapy-naive elderly patients (aged at least 76 years) with locally advanced or metastatic non-small-cell lung cancer were accrued. Eligible patients received at least two cycles of docetaxel at a dose of 60 mg/m2 on day 1 over 1 h every 3 weeks. Patients who were considered ineligible for this study were also registered. Symptom control was assessed using a questionnaire during the treatment period. The pharmacokinetics of docetaxel were evaluated in the first cycle of chemotherapy. RESULTS: Of 35 elderly patients, 15 (43%) met the study eligibility criteria. The reasons for ineligibility consisted mainly of poor performance status, poor bone marrow function, and hypoxemia (six patients each). A total of 49 cycles of chemotherapy (median 2 cycles, range 1-12 cycles) were administered to the eligible patients, six of whom achieved a partial response (overall response rate 40%, 95% confidence interval 15-65%). The major toxicity was hematologic, with grade 3 or greater neutropenia and grade 3 neutropenic fever developing in 13 patients (87%) and five patients (33%), respectively. Symptoms, as assessed in terms of the symptom control score, did not clearly decline during the treatment period. The values (mean+/-SD) of Cmax, AUC(0-->inf), and t(1/2) were 1.35+/-0.32 microg/ml, 1.79+/-0.52 microg h/ml, and 4.1+/-2.3 h, respectively. CONCLUSIONS: Although the validity of the results of this study is limited due to the small sample size, docetaxel appears effective in selected elderly patients with advanced non-small-cell lung cancer. PMID- 15127233 TI - Chronic eosinophilic leukemia with 46,XY,1,der(1;7)(q10;p10) translocation. PMID- 15127234 TI - Anatomical histological and mesoscopic study of the adipose tissue of the orbit. AB - Our study aimed to define the organization of the orbital adipose tissue, which is constituted from white adipose tissue. Six orbital samples were taken by dissection from fresh cadavers. After fixation and paraffin-embedding, the blocks were sectioned in the three spatial planes (two in the frontal, two in the sagittal, two in the horizontal). Semi-serial sections of 7 microm were then stained with hematein, eosin, safran or Masson trichrome green. We noticed strong areas of adhesion with orbital bones located at the lacrimal gland, the orbital trochlea and the inferior orbital fissure. Our mesoscopic and histological results allowed the description of two types of orbital adipose tissue corresponding to morpho-functional topographic variations. One was constituted of thick conjunctival septa with small adipocytes near muscles and the lacrimal gland. This was a supporting tissue that gave the points of rotation. The other was constituted of thin conjunctival septa with larger adipocytes near the optic nerve, allowing its movements in the orbit. These morphological differences appeared to be correlated with the mechanical role of these two areas. The dense appearance could correspond to the functional trochlea of rectus muscles described. In contrast we did not observe the systematic radial and concentric conjunctival meshwork classically described. This study underlines the specificity of orbital adipose tissue, which could be useful for a better understanding of its normal and pathological partition and its involvement in ocular motility. PMID- 15127235 TI - Expression of NK-associated receptors on cytotoxic T cells from melanoma patients: a two-edged sword? AB - The coexistence of tumor progression with a tumor-specific immune response constitutes a major paradox of tumor immunity. During the last decade, the presence of cytotoxic T lymphocytes (CTLs) recognising melanoma-associated antigens has been unequivocally demonstrated in numerous different in vivo and in vitro models. However, most often these melanoma-specific T lymphocytes do not control tumor growth. Several mechanisms that involve changes in melanoma phenotype and/or in T-cell differentiation and function could explain the inability of the immune response to control melanoma. In the last few years it has been demonstrated that cellular cytotoxicity is the result of a balance between activating signals triggered by the TCR and costimulatory molecules and inhibitory signals triggered by inhibitory receptors expressed by the CTL. Because the final outcome of the immune response against melanoma depends on the balance between activating and inhibitory signals, the expression de novo on melanoma cells of ligands for inhibitory NKRs and the down-regulation of costimulatory molecules may favor the escape of tumor cells from immunosurveillance. In this paper we review how altered expression of molecules required for T-cell costimulation could result in impaired lysis of melanoma. The modulation of antimelanoma T-cell responses by a group of receptors originally described on NK cells (NK-associated receptors) but which are now known also to be expressed on a subset of cytolytic effector cells is reviewed. We hypothesize that the expression of ligands for NKRs on melanoma cells may contribute to T cell-mediated immune responses against melanoma either enhancing or inhibiting activation and differentiation to effector cells. Blocking inhibitory receptors or increasing activating receptors could result in new strategies to improve T cell-mediated rejection of melanoma. PMID- 15127236 TI - A phase I trial of idiotypic vaccination with HMFG1 in ovarian cancer. AB - INTRODUCTION: An extended phase I trial was conducted in a total of 26 patients with ovarian cancer. The objectives were to assess the safety and tolerability of idiotypic vaccination using the murine monoclonal antibody HMFG1 (anti-MUC1), and to develop robust assays to monitor humoral immune responses generated against either the antibody or MUC1. MATERIAL AND METHODS: All patients had undergone standard debulking surgery (where appropriate) and at least one regimen of platinum-based chemotherapy. Eligibility criteria included: (a) residual disease at the end of chemotherapy, (b) relapsed disease, and (c) pathologically confirmed second complete remission following salvage chemotherapy. Patients received a priming dose of 25 mg of HMFG1 either intravenously or intraperitoneally, followed by up to six intradermal doses of HMFG1 in 10% Alhydrogel at intervals of 1 month. The three dose levels were 0.5 mg, 1 mg and 5 mg. We devised modifications of published protocols for the measurement of anti idiotypic and anti-MUC1 antibody responses and also extended the use of the IAsys resonant mirror biosensor to measure the kinetics of the idiotypic network response in these patients. RESULTS: There were no serious adverse events at any dose level. The trial confirmed that all doses could be administered safely with minimal toxicity. No clinical responses were seen in patients with evaluable disease. ELISA for anti-idiotypic antibodies (Ab2) showed significant levels in patients who completed the protocol. There were no significant differences in the levels of Ab2 generated by the different doses of antibody. These results were confirmed by biosensor assays for Ab2, which also showed affinity maturation of the Ab2 response as patients progressed through the vaccination protocol. Biosensor assays also demonstrated no difference in the affinity of Ab2 generated by different booster doses of HMFG1. ELISA for anti-MUC1 antibodies showed less consistent results, with very small but statistically significant rises in anti MUC1 signals seen in 38% of patients who completed the vaccination regimen. DISCUSSION: The clinical endpoints of safety and tolerability were met. The assays developed for this project have shown reproducibility and may provide surrogate endpoints to assess vaccination for future trials. The use of similar biosensors may be of particular relevance for monitoring of humoral immune responses in other vaccine trials. The low levels of anti-MUC1 antibodies generated may correspond with the lack of clinical efficacy in the few patients with evaluable disease. PMID- 15127237 TI - Heat shock protein 70 / MAGE-1 tumor vaccine can enhance the potency of MAGE-1 specific cellular immune responses in vivo. AB - The cancer-testis antigen encoded by the MAGE-1 gene is an attractive antigen in tumor immunotherapy because it can be processed as a foreign antigen by the immune system and generate tumor-specific cellular immune response in vivo. However, increase of the potency of MAGE-1 DNA vaccines is still needed. The high degree of sequence homology and intrinsic immunogenicity of heat shock protein 70 (HSP70) have prompted the suggestion that HSP70 might have immunotherapeutic potential, as HSP70 purified from malignant and virally infected cells can transfer and deliver antigenic peptides to antigen-presenting cells to elicit peptide-specific immunity. In this research, we evaluated the enhancement of linkage of Mycobacterium tuberculosis HSP70 to MAGE-1 gene of the potency of antigen-specific immunity elicited by naked DNA vaccines. We found that vaccines containing MAGE-1-HSP70 fusion genes enhanced the frequency of MAGE-1-specific cytotoxic T cells in contract to vaccines containing the MAGE-1 gene alone. More importantly, the fusion converted a less effective DNA vaccine into one with significant potency against established MAGE-1-expressing tumors. These results indicate that linkage of HSP70 to MAGE-1 gene may greatly enhance the potency of DNA vaccines, and generate specific antitumor immunity against MAGE-1-expressing tumors. PMID- 15127238 TI - Immunotherapy with histamine and interleukin 2 in malignant melanoma with liver metastasis. PMID- 15127239 TI - Circulating tumour markers in breast cancer. AB - A large number of markers have been proposed for breast cancer, but among them only CA 15.3, CEA and cytokeratins (i.e. TPA, TPS and Cyfra 21.1) are currently used in clinical practice. Serum marker levels reflect tumour burden and for this reason they are not sensitive enough to be used for screening and early diagnosis of primary breast cancer. By contrast, the role of tumour markers is established in the diagnosis of recurrent disease and in the evaluation of response to treatment. In the former case, however, prospective randomised studies are required to demonstrate any survival benefit when earlier therapeutic interventions are instituted upon elevation of serum markers. In the second case, tumour marker evaluation represents a simple, objective method for monitoring of therapeutic response that seems to offer significant advantages over conventional imaging methods (e.g. objectivity, modifications in tumour biology). Furthermore, research studies are ongoing to identify and validate new biochemical parameters which can be of use not only in advanced disease but also in other stages of the diagnostic work-up of breast cancer. PMID- 15127240 TI - Biological characterisation of breast cancer by means of PET. AB - Breast cancer is associated with increased glucose consumption and can therefore be visualised with the glucose analogue [(18)F]2-deoxy-2-fluoro-D-glucose (FDG) and positron emission tomography (PET). FDG uptake in the primary tumour can vary substantially, and specific tumour characteristics have been demonstrated to determine the degree of glucose metabolism. Factors with a major influence on FDG uptake in breast cancer comprise expression of glucose transporter Glut-1 and hexokinase I, number of viable tumour cells per volume, histological subtype, tumour grading, microvessel density and proliferative activity. Recently, an association between high FDG uptake and a worse prognosis was suggested. Several studies have been performed correlating FDG uptake with a variety of prognostic and molecular biomarkers as well as parameters predicting tumour response to therapy. However, a correlation with important clinical prognostic markers such as axillary lymph node status and size of the primary tumour, expression of oestrogen and progesterone receptors, proto-oncogene c-erbB-2 or VEGF could not be demonstrated. The lack of correlation with important markers of prognosis does not suggest that FDG uptake might be used as a prognostic criterion in breast cancer. Innovative radiotracers for specific imaging of tumoural perfusion ([(15)O]H(2)O), hormone receptor expression ([(18)F]FES), protein synthesis ([(11)C]methionine), proliferation rate ([(18)F]FLT) or bone mineralisation ([(18)F]fluoride) may provide additional information compared with that provided by FDG PET. PMID- 15127241 TI - 153Sm-EDTMP for bone pain palliation in skeletal metastases. AB - 153Sm-ethylene diamine tetramethylene phosphonate (EDTMP) is a widely available and extensively tested radiopharmaceutical for systemic radionuclide therapy in patients with symptomatic multiple skeletal metastases. Its use is approved for any secondary bone lesion which has been shown to accumulate (99m)Tc-methylene diphosphonate, including breast carcinoma. The molecule is stable in vitro and upon injection more than 50% of the dose is avidly fixed by lesional and non lesional bone, with the rest being rapidly eliminated unchanged via the urine. The short half-life (46.3 h), the relatively low-energy beta emissions (E(ave)=233 keV) and the gamma emission (103 keV) make (153)Sm a very attractive radionuclide, allowing therapeutic delivery of short-range electrons at relatively high dose rates with external imaging to corroborate biodistribution and possible dosimetric estimates. For a standard dose of 2,590 MBq/70 kg, the estimated radiation dose to metastases is 86.5 Gy. Critical organs are the bladder wall (2.5 Gy/2,590 MBq) and red marrow (4 Gy/2,590 MBq), with the latter being the critical factor in clinical practice as the dose-limiting factor is marrow radiotoxicity. The therapy has, however, proved safe provided that the platelet count exceeds 100 x 10(9)/l and the white blood cell count exceeds 3.5 x 10(9)/l. Clinical data obtained in fewer than 250 patients, within several studies, lead to the following conclusions: a dose of 37 MBq/kg has a better therapeutic ratio than a dose of 18.5 MBq/kg; the mean pain palliation rate after a single treatment in breast cancer is about 80%; toxicity is generally mild and transitory; and re-treatments are effective and safe provided that haematological values have fully recovered. PMID- 15127243 TI - Calcific haemorrhagic bursitis anterior to the knee mimicking a soft tissue sarcoma: report of two cases. AB - We describe the radiological and pathological findings of two cases of calcific haemorrhagic bursitis, one involving the superficial infrapatellar bursa and the other the prepatellar bursa. It was the presence of dystrophic calcification within the lesion that suggested a mineralizing soft tissue sarcoma such as synovial sarcoma. As the radiographic and MR features of the two conditions can be similar but the appropriate management very different, rare calcifying haemorrhagic bursitis needs to be included in the differential diagnosis of masses adjacent to the knee joint showing calcification. PMID- 15127242 TI - The choice of the correct imaging modality in breast cancer management. AB - This brief overview discusses which of the diagnostic options are more reliable and effective for breast cancer imaging with a view to avoiding the unjustified use of techniques that are suboptimal. The technological development of diagnostic imaging has been very impressive, and both radiological (mammography, ultrasonography, computed tomography, magnetic resonance imaging) and nuclear medicine tools (bone scan, planar and SPECT scintigraphy, sentinel node biopsy, positron emission tomography) have helped to overcome past limitations in the detection of small lesions. Furthermore, new approaches have been developed that permit successful differential diagnosis of doubtful lesions and rapid identification of systemic metastases, and allow non-invasive characterisation of the biology of cancer tissue. There is evidence that these advances may have helped in optimising therapeutic strategies. Importantly, the metabolic information provided by nuclear medicine procedures may be combined with the anatomical data supplied by radiological techniques in order to assist in predicting tumour response, planning radiotherapy and monitoring patient outcome. It is difficult to formulate conclusive diagnostic guidelines for application in the work-up of breast cancer, because while the role of some examinations, such as mammography and ultrasonography, is well established, that of others, such as magnetic resonance imaging and positron emission tomography, is still a matter of debate. There is a need for further prospective evaluations with appropriate clinical trials designed to evaluate the impact of these approaches in improving survival and quality of life. PMID- 15127244 TI - Musculoskeletal disorders associated with HIV infection and AIDS. Part II: non infectious musculoskeletal conditions. AB - This section of a two-part series on musculoskeletal disorders associated with HIV infection and AIDS reviews the non-infectious musculoskeletal conditions. In the first part, the infectious conditions were reviewed. The non-infectious conditions include polymyositis, drug-induced myopathy, myositis ossificans, adhesive capsulitis, avascular necrosis, bone marrow abnormalities, and hypertrophic osteoarthropathy. Inflammatory and reactive arthropathies are more prevalent in HIV-positive individuals, and a separate section is dedicated to these conditions, including Reiter's syndrome, psoriatic arthritis, HIV associated arthritis, painful articular syndrome, and acute symmetric polyarthritis. Lastly, we include a discussion of HIV-related neoplastic processes that affect the musculoskeletal system, namely Kaposi's sarcoma and non Hodgkin's lymphoma. PMID- 15127245 TI - First report of an accessory popliteal muscle: detection with MRI. AB - During an MRI examination of the knee in a 48-year-old patient suffering from degenerative changes of a partly resected medial meniscus and concomitant osteoarthritis of the knee joint, an unusual variant of an accessory muscle in the popliteal fossa was found. To our best knowledge this muscle has never been described before. Because of the close relationship to the popliteal muscle with regard to course and localisation in the deep popliteal fossa ventral to the popliteal artery, the term "accessory popliteal muscle" is proposed. PMID- 15127246 TI - Recurrent Mycobacterium marinum tenosynovitis of the wrist mimicking extraarticular synovial chondromatosis on MR images. AB - Tenosynovitis caused by atypical mycobacterial infections may produce rice bodies within affected tendon sheaths. We report a case of recurrent M. marinum infection involving the flexor tendons within the carpal tunnel in which the rice bodies were mistaken for synovial chondromatosis on MR images. PMID- 15127247 TI - Skeletal muscle metastasis from uterine leiomyosarcoma. AB - A case of a 68-year-old woman who presented with a rapidly enlarging painful right thigh mass is presented. She had a known diagnosis of uterine leiomyosarcoma following a hysterectomy for dysfunctional uterine bleeding. She subsequently developed a single hepatic metastatic deposit that responded well to radiofrequency ablation. Whole-body MRI and MRA revealed a vascular mass in the sartorius muscle and a smaller adjacent mass in the gracilis muscle, proven to represent metastatic leiomyosarcoma of uterine origin. To our knowledge, metastatic uterine leiomyosarcoma to the skeletal muscle has not been described previously in the English medical literature. PMID- 15127248 TI - Role of substrate on the conformational stability of the heme active site of cytochrome P450cam: effect of temperature and low concentrations of denaturants. AB - The effect of 1R-camphor on the conformational stability of the heme active site of cytochrome P450cam has been investigated. The absorption spectra of the heme moiety showed the presence of two hitherto unknown intermediates formed at low urea concentrations or during small temperature perturbations. The corresponding thermodynamic parameters were obtained by global fitting of the experimental data to a generalized sequential unfolding model at different wavelengths, which showed that the active conformation of the enzyme is stabilized by binding of the substrate at the active site. Circular-dichroism spectra of the enzyme in the visible- and far-UV region were studied to identify the critical range of denaturant concentration and the temperature at which the tertiary structure around the heme center was affected with almost no change in the secondary structure of the enzyme. This critical range of urea concentration was 0-2.8 M in the presence of camphor and 0-1.5 M in the absence of camphor. The tertiary structure of the enzyme was found to undergo conformational change in the temperature range 20-60 degrees C in the presence of the substrate and 20-47 degrees C in its absence. The spectral assignments of the intermediate species of the heme active site with the intact secondary structure of the enzyme were made by deconvolution of the Soret absorption spectra, and the results were analyzed to determine stabilization of the heme active-site geometry by 1R-camphor. Results showed that subtle conformational changes due to melting of the tertiary contacts in the active site lead to formation of intermediates which are coordinatively similar to the native enzyme. Analogous intermediate species might be responsible for leakage in the redox catalytic cycle of the enzyme. PMID- 15127249 TI - Determination of reduction potential of an engineered Cu(A) azurin by cyclic voltammetry and spectrochemical titrations. AB - The reduction potentials of an engineered CuA azurin in its native and thermally denatured states have been determined using cyclic voltammetry and spectrochemical titrations. Using a 4,4'-dipyridyl disulfide modified gold electrode, the reduction potentials of native and thermally denatured CuA azurin are the same within the experimental error (422 +/- 5 and 425 +/- 5 mV vs. NHE, respectively, in 50 mM ammonium acetate buffer, pH 5.1, 300 mM NaCl, 25 degrees C), indicating that the potential is that of a nonnative state. In contrast, using a didodecyldimethylammonium bromide (DDAB) film-pyrolytic graphite edge (PGE) electrode, the reduction potentials of native and thermally denatured CuA azurin have been determined to be 271 +/- 7 mV (50 mM ammonium acetate buffer, pH 5.1, 4 degrees C) and 420 +/- 1 mV (50 mM ammonium acetate buffer, pH 5.1, 25 degrees C), respectively. Spectroscopic redox titration using [Ru(NH3)5Py]2+ resulted in a reduction potential (254+/-4 mV) (50 mM ammonium acetate buffer, pH 5.1, 4 degrees C) similar to the value obtained using the DDAB film-PGE electrochemical method. Complete reoxidation of [Ru(NH3)5Py]2+-reduced CuA azurin is also consistent with the conclusion that this spectrochemical titration method using [Ru(NH3)5Py]2+ measures the reduction potential of native CuA azurin. PMID- 15127250 TI - Tranexamic acid for major spinal surgery. AB - Patients who undergo major spinal surgery often require multiple blood transfusions. The antifibrinolytics are medications that can reduce blood transfusion requirements in cardiac surgery and total knee arthroplasty. The present role of synthetic antifibrinolytics, especially tranexamic acid, in reducing peri-operative blood-transfusion requirements in spine surgery is still unclear. The majority of studies exploring the role of these drugs in spine surgery have limited patient enrollment and report mixed results. The goal of the present review is to discuss the pharmacology of tranexamic acid briefly. A brief synopsis of the studies using the synthetic antifibrinolytics for spine surgery is presented. Finally, the potential risks and the benefits of antifibrinolytics are discussed. PMID- 15127252 TI - Volatile anesthetic antagonism by long-chain free fatty acids. PMID- 15127251 TI - Reviewer's comment regarding MS-no: 03/057 entitled: "Kyphoplasty for treatment of osteoporotic vertebral fractures: a prospective non-randomized study" by U. Berlemann et al. PMID- 15127253 TI - Changes in body temperature during profound hypothermic cardiopulmonary bypass in adult patients undergoing aortic arch reconstruction. AB - PURPOSE: Our aim was to characterize changes in body temperatures during profound hypothermic cardiopulmonary bypass (CPB) conducted with the sternum opened. METHODS: In ten adult patients who underwent profound hypothermic (< 20 degrees C) CPB for aortic arch reconstruction, pulmonary arterial temperature (PAT), nasopharyngeal temperature (NPT), forehead deep-tissue temperature (FHT), and urinary bladder temperature (UBT) were recorded every 1 min throughout the surgery. In addition, the CPB venous line temperature (CPBT), a reasonable indicator of mixed venous blood temperature during CPB and believed to best reflect core temperature during stabilized hypothermia on CPB, was recorded during the period of total CPB. RESULTS: PAT began to change immediately after the start of cooling or rewarming, closely matching the CPBT ( r = 0.98). During either situation, the other four temperatures lagged behind PAT ( P < 0.05); however, NPT followed PAT more closely than the other three temperatures ( P < 0.05). During stabilized hypothermia, PAT, NPT, and FHT, but not UBT, closely matched the CPBT, with gradients of less than 0.5 degrees C. CONCLUSION: During induction of profound hypothermia and its reversal on total CPB with the heart in situ, a PA catheter thermistor, presumably because of its placement immediately behind the superior vena cava, would provide a reliable measure of the mixed venous blood temperature. During stabilized profound hypothermia, PAT, NPT, and FHT, but not UBT, serve as a reliable index of core temperature. PMID- 15127254 TI - Effects of withdrawal of phasic lung inflation during normocapnia and hypercapnia on the swallowing reflex in humans. AB - PURPOSE: This study was done to test the hypothesis that hypercapnia has a direct, inhibitory effect on swallowing. METHODS: We investigated changes in the frequency and timing of repeated swallows induced by continuous infusion of water into the pharynx before, during, and after transient airway occlusion at normocapnia and hypercapnia in 12 healthy volunteers. Hypercapnia was induced by adding a dead space. Ventilation was monitored using a pneumotachograph, and swallowing was identified by submental electromyogram. RESULTS: We found that hypercapnia decreased the frequency of swallows (8.2 +/- 3.7 vs 11.4 +/- 5.3 swallows.min-1 [mean +/- SD]: hypercapnia vs normocapnia; P < 0.05), together with a loss of the preponderant coupling of swallows with expiratory phase observed at normocapnia. We also found that the withdrawal of phasic lung inflation produced by airway occlusion at end-expiration suddenly increased the swallowing frequency, both at normocapnia (from 11.4 +/- 5.3 to 16.7 +/- 3.7 swallows.min-1; P < 0.01) and at hypercapnia (from 8.2 +/- 3.7 to 22.0 +/- 6.7 swallows.min-1; P < 0.01). Although the degree of increased swallowing frequency during airway occlusion was more prominent at hypercapnia than at normocapnia ( P < 0.05), the distribution of the timing of swallows in relation to the phase of the respiratory cycle during airway occlusion at hypercapnia was similar to that during airway occlusion at normocapnia. CONCLUSION: The results of our study strongly suggest that the attenuation of the swallowing reflex during hypercapnia is not due to the direct, inhibitory effect of CO2 on the swallowing center, but, rather, is due to the increased inhibitory influence of a lung-volume-related reflex. PMID- 15127256 TI - Multi-Unit and Multi-Path system of the neural network can explain the steep dose response of MAC. AB - PURPOSE: The slope of the dose-response curves of inhalation anesthetics is steep around the minimum alveolar concentration of inhalation anesthetics (MAC) value. Contrastingly, the anesthetic dose-response curves of ion channels and enzymes are gradual. This discrepancy in the steepness may be a key to solve the mechanisms of anesthesia. To explain the steepness we propose a mathematical model of the neural network related to MAC. METHODS: We assumed that, in order to show movement in response to a noxious stimulus, a signal needed to be transmitted from A to B. There are m conduction pathways (Multi-Path) in the nerve network between A and B, and there are n conduction units (Multi-Unit) in each conduction pathway. Anesthetics bind to each conduction unit and block signal transmission. Anesthetics prevent movement in response to a stimulus, when at least one conduction unit among all conduction pathways has been blocked. We derived the equation for the probability of the signal being blocked by anesthetics. RESULTS: The steep dose-response curve of in vivo anesthesia requires a very large number of conduction units ( n > 100) and conduction pathways ( m > 10(6)). The EC50 for each conduction unit was at least 3.8-fold larger than the apparent EC50 for the whole system under the experimental condition of simulation. CONCLUSIONS: We constructed a model for the neural networks that relates to MAC as a Multi-Unit and Multi-Path system (MUMPS). To obtain highly cooperative dose-response curves comparable to those of in vivo anesthesia, at least 10(6) conduction pathways and more than 100 conduction units are required for each pathway. In these systems, the apparent anesthetic potency on the whole system (MAC) is much stronger than the anesthetic action on each unit. Because of this discrepancy, it is important to set anesthetic concentrations appropriately for experiments with in vitro systems. PMID- 15127255 TI - Antagonizing potencies of saturated and unsaturated long-chain free fatty acids to isoflurane in goldfish. AB - PURPOSE: We have previously reported that myristate, a saturated free fatty acid (FFA) with 14 carbons (C14), antagonizes volatile anesthetics in goldfish. The hydrophobicity and molecular configuration of FFAs may play an important role in the antagonizing effect. To examine their contribution, we investigated the antagonizing potencies of saturated and unsaturated long-chain FFAs in goldfish. METHODS: Saturated and monounsaturated FFAs of C14-18 were tested. We determined the anesthetic concentration producing a 50% effect (EC50) of isoflurane in the absence or presence of FFA by observing the escape reaction of goldfish against an electrical stimulus. RESULTS: All FFAs increased the EC50 of isoflurane dose dependently compared with reactions in the absence of FFA ( P < 0.05). For saturated FFAs, the relationship between chain lengths and antagonizing potencies was not linear. C18 was the most effective and C16 was the least effective antagonist ( P < 0.05). Among unsaturated FFAs, C14 was the most effective antagonist ( P < 0.05). In a comparison of saturated and unsaturated FFAs, saturated C14 and C18 were more effective antagonists than unsaturated FFAs of the same carbon numbers ( P <<0.05). CONCLUSION: The hydrophobicity of FFAs increases as the chain length increases. Therefore, our findings suggest that the antagonizing effect of long-chain FFAs in goldfish, in terms of their capacity to perturb the lipid membrane structure, may be determined not solely by their hydrophobicity but also by their molecular configuration. PMID- 15127258 TI - Toborinone and olprinone, phosphodiesterase III inhibitors, inhibit human platelet aggregation due to the inhibition of both calcium release from intracellular stores and calcium entry. AB - PURPOSE: We investigated the inhibitory effects of toborinone and olprinone on human platelet aggregation and calcium mobilization.Abstract Copyright: METHODS: Washed human platelets were preincubated with toborinone or olprinone, then exposed to 0.015 U.ml-1 of thrombin. Aggregation curves were measured using an aggregometer. Effects of toborinone or olprinone on changes in intracellular calcium concentration ([Ca2+]i) were measured fluorometrically using fura-2 acetoxymethyl ester (fura-2). Levels of intracellular cyclic 3",5"-adenosine monophosphate concentration ([cAMP]i) were also measured, using enzyme-linked immunosorbent assay (ELISA) techniques. RESULTS: The concentrations required to cause 50% inhibition of aggregation (IC50) induced by thrombin were 9.7 +/- 0.9 micro M for toborinone and 3.6 +/- 0.2 micro M for olprinone. Both drugs at IC50 significantly elevated [cAMP]i levels and significantly inhibited Ca2+ release from intracellular stores. Release of [Ca2+]i induced by thrombin was 272.9 +/- 87.1 nM, 153.3 +/- 28.7 nM, and 138.9 +/- 58.2 nM in the control, toborinone, and olprinone groups, respectively ( P < 0.02). Calcium influx through calcium channels in the plasma membrane was also suppressed by toborinone and olprinone. CONCLUSION: Toborinone (9.7 micro M) and olprinone (3.6 micro M) inhibit human platelet aggregation, though these concentrations are higher than their therapeutic plasma concentrations. The inhibitory effects of both drugs are related to the inhibition of both Ca2+ release and Ca2+ entry through [cAMP]i elevation. PMID- 15127257 TI - Single sodium channels from human skeletal muscle in planar lipid bilayers: characterization and response to pentobarbital. AB - PURPOSE: To investigate the response to general anesthetics of different sodium channel subtypes, we examined the effects of pentobarbital, a close thiopental analogue, on single sodium channels from human skeletal muscle and compared them to existing data from human brain and human ventricular muscle channels. METHODS: Sodium channels from a preparation of human skeletal muscle were incorporated into planar lipid bilayers, and the steady-state behavior of single sodium channels and their response to pentobarbital was examined in the presence of batrachotoxin, a sodium-channel activator. Single-channel currents were recorded before and after the addition of pentobarbital (0.34-1.34 mM). RESULTS: In symmetrical 500 mM NaCl, human skeletal muscle sodium channels had an averaged single-channel conductance of 21.0 +/- 0.6 pS, and the channel fractional open time was 0.96 +/- 0.04. The activation midpoint potential was -96.2 +/- 1.6 mV. Extracellular tetrodotoxin blocked the channel with a half-maximal concentration (k1/2) of 60 nM at 0 mV. Pentobarbital reduced the time-averaged conductance of single skeletal muscle sodium channels in a concentration-dependent manner (inhibitory concentration 50% [IC50] = 0.66 mM). The steady-state activation was shifted to more hyperpolarized potentials (-16.7 mV at 0.67 mM pentobarbital). CONCLUSION: In the planar lipid bilayer system, skeletal muscle sodium channels have some electrophysiological properties that are significantly different compared with those of sodium channels from cardiac or from central nervous tissue. In contrast to the control data, these different human sodium channel subtypes showed the same qualitative and quantitative response to the general anesthetic pentobarbital. The implication of these effects for overall anesthesia will depend on the role the individual channels play within their neuronal networks, but suppression of both central nervous system and peripheral sodium channels may add to general anesthetic effects. PMID- 15127259 TI - Spinal neurotoxicity and tolerance after repeated intrathecal administration of YM 872, an AMPA receptor antagonist, in rats. AB - PURPOSE: Although the Alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) receptor antagonist, YM 872, has been considered to be useful in analgesia for both acute and chronic pain, there are no studies of its neurotoxicity and tolerance. We examined the spinal neurotoxicity and tolerance of YM 872 analgesia by repeated intrathecal administration in rats. METHODS: Male Sprague-Dawley rats with lumbar intrathecal catheters received YM 872 at 1 micro g. 10 micro l-1 (eight rats; YM group) or normal saline 10 micro l (eight rats; C group) intrathecally once a day for 30 days. We evaluated the analgesic effects every 3 days, by tail-flick test and behavioral side effects. On the 31st day, the lumbar spinal cord was removed from four randomly selected rats in each group for histological examination. RESULTS: The YM group showed significantly longer tail flick latency when subjected to a high-intensity light beam than the C group at each measurement time point, although no significant changes in the latency according to the time course of the study were observed for the entire study period of 30 days in either group. No rats showed any side effects. Histologically, only slight lymphocytic cell infiltration and degeneration of myelinated fibers occurred, similarly in both groups. No changes were observed in the spinal cord in either group. CONCLUSION: Administration of YM 872 (1 micro g) once a day for 30 days did not induce any tolerance and caused no histological changes in the spinal cord. PMID- 15127260 TI - Monitoring magnesium to guide magnesium therapy for heart surgery. PMID- 15127261 TI - Severe Legionella pneumophila pneumonia associated with the public bath on a cruise ship in Japan. PMID- 15127262 TI - Brachial plexus injury related to improper positioning during general anesthesia. PMID- 15127263 TI - Successful management of a patient with neuroleptic malignant syndrome associated with marked elevation of serum creatine kinase. PMID- 15127264 TI - Pneumothorax associated with epidural anesthesia. PMID- 15127265 TI - Integration of suppression ratio in the bispectral index. PMID- 15127266 TI - Modified long Trachlight wand for a double-lumen endobronchial tube. PMID- 15127267 TI - Pulmonary embolism after minor surgery in a patient with low-risk thrombocythemia. PMID- 15127268 TI - A new scoring system to classify patients with colorectal liver metastases: proposal of criteria to select candidates for hepatic resection. AB - BACKGROUND: Although many studies have reported the beneficial effects of hepatic resection for colorectal liver metastases on survival rates, it is still difficult to preoperatively select good candidates for hepatectomy. METHODS: Fifteen clinicopathological features, which were recognized only before or during surgery, were selected retrospectively in 81 consecutive patients in one hospital (Group I). These features were entered into a multivariate analysis to determine independent and significant variables affecting long-term prognosis after hepatectomy. Using selected variables, we created a scoring formula to classify patients with colorectal liver metastases to select good candidates for hepatic resection. The usefulness of the new scoring system was examined in a series of 70 patients from another hospital (Group II). RESULTS: Multivariate analysis, i.e., Cox regression analysis, showed that serosa invasion of primary cancers (P = 0.0720, risk ratio = 2.238); local lymph node metastases of primary cancers, i.e., Dukes' C (P = 0.0976, risk ratio = 2.311); multiple nodules of hepatic metastases (P = 0.0461, risk ratio = 2.365); nodules of hepatic metastases greater than 5 cm in diameter (P =0.0030, risk ratio = 4.277); and resectable extrahepatic distant metastases (P = 0.0080, risk ratio = 4.038) were significant and independent prognostic factors for poor survival after hepatectomy. Using these five variables, we created a new scoring formula to classify patients with colorectal liver metastases. Finally, our new scoring system classified patients in Group II and Group I well, according to long-term outcomes after hepatic resection. CONCLUSIONS: Our new scoring system to classify patients with colorectal liver metastases is simple and useful in the preoperative selection of good candidates for hepatic resection. Although many studies have reported the beneficial effects of hepatic resection for colorectal liver metastases on survival rates, it is still difficult to preoperatively select good candidates for hepatectomy. PMID- 15127269 TI - Surgery for multiple hepatic colorectal metastases. AB - The purpose of this review is to address three important questions concerning hepatic resection for multiple colorectal metastases. (1) Is the number of tumors truly a significant prognostic factor? (2) Are patients with four or more tumors contraindicated for hepatic resection? (3) Up to how many nodules should we attempt to resect? Although the efficacy of surgical resection for one to three hepatic metastases is clear, based on several reports, the literature regarding the resection of four or more metastatic lesions is conflicting. Review of the data at our institutions showed that the number of tumors was a significant prognostic factor, because patient survival after liver resection for multiple metastases was worse than that for single metastasis. However, patients with two or three nodules and those with four or more nodules showed the same survival curves, or those with four or more metastases fared even better. Therefore, patients with four or more metastases should be considered for hepatic resection. The maximum number of hepatic tumors in longterm survivors reported in the literature has been increasing, and the limit for the number of respectable metastases has not yet been determined. Because liver resection is still the only treatment that offers a cure, surgery for multiple metastases may be justified as long as the operation is safe and technically feasible. PMID- 15127270 TI - Extensive resections for colorectal liver metastases. AB - BACKGROUND/PURPOSE: Mortality and morbidity rates after liver resections have decreased with better surgical techniques and perioperative care. The aim of this study was to evaluate the short- and longterm results in patients who had undergone extensive hepatectomies. METHODS: From January 1985 to December 2000, 237 patients underwent 275 liver resections for colorectal metastases. Extensive liver resections were defined as follows: technical reasons (extended hepatectomies, associated vascular resections); disease extent (diameter, >>10 cm; number, >>5; associated extrahepatic resection). The total number of extensive liver resections was 74. There were 51 radical resections (68.9%), while in the nonextensive resections group, 152 resections were radical (90.7%; P = 0.1). RESULTS: Postoperative mortality (60 days) was 1.6% (1.3% in the extensive resections group; P = 0.3), while morbidity was 22.7% (31% in the extensive resections group vs 19% in the nonextensive resections group; P = 0.1). One-, 3-, and 5-year overall actuarial survival rates were 91.8%, 44.9%, and 25.3%. The survival rates of patients who underwent an extensive resection were similar to those in the nonextensive resections group. CONCLUSIONS: Technical difficulties and neoplastic extension are not, nowadays, a contraindication for hepatectomy for colorectal liver metastases, unless a radical resection is performed. Mortality and morbidity rates after liver resections have decreased with better surgical techniques and perioperative care. The aim of this study was to evaluate the short- and longterm results in patients who had undergone extensive hepatectomies. PMID- 15127271 TI - Local ablation for unresectable liver tumors: is thermal best? AB - Hepatic resection remains the "gold standard" for patients with resectable disease. Nevertheless, for a variety of reasons this is not feasible for the majority of patients. A wide range of locally ablative techniques has been developed for use in these patients with the aim of improving survival. Unfortunately, as with many recent techniques in surgery, much of the development of these methods, and particularly their introduction clinically, has not been based on sound scientific data. The relative merits and limitations of the more commonly used techniques are discussed, although this lack of prospective, randomized data precludes firm conclusions to be drawn from many of the studies reported. By far the most popular methods now employed, thermal techniques have certain limitations, particularly when treating tumors adjacent to major vascular or biliary structures. The authors believe that this situation represents the "niche" for which ablative techniques may ultimately find their logical application, where a single awkwardly placed metastasis deems a patient unresectable. If such a metastasis can be completely and safely ablated, a potentially curative resection may then become a realistic option. The relatively new, nonthermal technique of hepatic electrolysis has been extensively studied and shown to be safe and effective in close proximity to major intrahepatic veins due to a subtle electrochemical action rather than a rapid "burn". This technique is discussed in the context of other, more traditional thermal methods of ablation. PMID- 15127272 TI - Applications of minimally invasive surgery in the management of inflammatory and neoplastic diseases of the pancreas. PMID- 15127273 TI - A gallbladder flap for reconstruction of the common bile duct. An experimental study on pigs. AB - BACKGROUND/PURPOSE: In this article we investigate whether a gallbladder flap could be used for reconstruction of the common bile duct (CBD), thereby providing drainage via an intact sphincter of Oddi. METHODS: Eight LWD pigs were used for the experiments. The gallbladder was dissected from its fossa, care being taken not to damage its vessels. The CBD was then displayed and approximately 5 mm resected. In two pigs a tube, and in six pigs a sphere, was constructed from the gallbladder flap. Anastomoses were constructed between the gallbladder flap and the CBD. Blood samples were drawn and on day 10 a laparotomy and an intraoperative cholangiography was performed. RESULTS: No pigs showed any sign of biliary leakage and standard liver parameters were not affected by surgery. In the two pigs who had a tube constructed, cholangiography showed extrahepatic stenosis and intrahepatic biliary dilatation. In the six pigs who had a sphere constructed, cholangiography was without any sign of extrahepatic stenosis or intrahepatic dilatation. CONCLUSION: Reconstruction of the CBD by way of a gallbladder flap seems to be a safe procedure to accomplish biliary drainage. This procedure could be of clinical significance for reconstruction of the CBD after resections for benign strictures and tumors, and for reconstruction after blunt or penetrating trauma. In this article we investigate whether a gallbladder flap could be used for reconstruction of the common bile duct (CBD), thereby providing drainage via an intact sphincter of Oddi. PMID- 15127274 TI - Laparoscopic right hemihepatectomy for a case of polycystic liver disease with right predominance. AB - BACKGROUND/PURPOSE: A case of polycystic liver disease with right predominance treated with laparoscopic right hemihepatectomy is described. PATIENT: A 43-year old woman complaining of right upper abdominal pain came in for consultation. Abdominal ultrasonography and computed tomography studies showed multiple liver cysts occupying mainly the right lobe, renal cysts, and splenomegaly. METHODS: Four trocars were used. A 12-mm trocar placed under the umbilicus was used for abdominal exploration. The other three trocars, two 12-mm trocars and one 5-mm trocar, were used as working ports. The liver was transected with ultrasound scissors and LigaSure. Major vessels such as the right portal vein, the right bile duct, and the hepatic vein were divided with a vascular endostapler. Operation time was 320 min, intraoperative blood loss was 120 ml, and postoperative oral intake occurred on day 3. RESULTS: No complication was observed during the perioperative period. CONCLUSIONS: Laparoscopic right hemihepatectomy is generally considered to cause excessive intraoperative bleeding and a long operation time. For our patient with multiple liver cysts, the procedure was a safe and minimally invasive option because little hepatic parenchymal resection was necessary for the multiple cysts.A case of polycystic liver disease with right predominance treated with laparoscopic right hemihepatectomy is described. PMID- 15127275 TI - Celiac axis stenosis in pancreaticoduodenectomy. AB - BACKGROUND/PURPOSE: Celiac axis (CA) stenosis is a fairly common condition in candidates for upper abdominal surgery. In this report, we have documented 5 patients with CA stenosis or occlusion who underwent pancreaticoduodenectomy, and we have discussed the surgical strategy for this condition. METHODS: We reviewed the records of 126 patients who had undergone resection of the pancreatic head during the past 10 years, and a search was made for obstructive lesions of the CA. RESULTS: Among these 126 patients, there were 5 cases (4%) of CA stenosis or occlusion. Three of them were interpreted as typical CA compression syndrome, based on arteriographic or operative findings, and an excision of the median arcuate ligament dramatically improved the blood flow of the celiac branches after division of the main collaterals. The remaining 2 patients, with organized occlusion of the CA, successfully underwent pancreaticoduodenectomy through preservation of the collateral pathway between the superior mesenteric artery and the celiac branch. In one of these 2 patients, the main collateral pathway, which ran across immediately behind the pancreatic head, was skeletonized and preserved. All 5 patients had uneventful postoperative courses. CONCLUSIONS: We conclude that, in pancreaticoduodenectomy for patients with organized CA occlusion, precise assessment of the vascular anatomy and preservation of the main collateral pathway are essential for carrying out a successful operation. Celiac axis (CA) stenosis is a fairly common condition in candidates for upper abdominal surgery. In this report, we have documented 5 patients with CA stenosis or occlusion who underwent pancreaticoduodenectomy, and we have discussed the surgical strategy for this condition. PMID- 15127276 TI - Comparison of urinary excretion of pravastatin and temocapril in bile duct ligated rats and Eisai hyperbilirubinemic rats (EHBR). AB - BACKGROUND/PURPOSE: In patients with complete bile duct obstruction, the only pathway for the elimination of cholephilic compounds is through the urine. Although changes in various transporters in the liver and kidney in cholestasis have been elucidated, little is known about how effectively the elimination of these compounds is compensated for by urinary excretion. METHODS: In the present study, the urinary excretion of pravastatin and temocapril was studied in bile duct-ligated rats (BDLR) for 3 days and in Eisai hyperbilirubinemic rats (EHBR). After urinary bladder cannulation, radiolabeled pravastatin and temocapril were injected intravenously. Urine samples were collected every 1 h for 4 h, and the radioactivity was counted. RESULTS: Urinary excretion of pravastatin was markedly increased in BDLR (85.9% of the dose after 4 h) and moderately increased in EHBR (35.9% of the dose after 4 h) compared with that in control rats (5.5% of the dose after 4 h). Similar but less prominent differences were observed with temocapril after it was administered (50.7%, 38.2%, and 22.0% of the dose after 4 h in BDLR, EHBR, and the controls, respectively). CONCLUSIONS: The absence of biliary excretion of anionic drugs was compensated for by urinary excretion in BDLR and EHBR, and the compensation was more efficient with pravastatin than with temocapril. In patients with complete bile duct obstruction, the only pathway for the elimination of cholephilic compounds is through the urine. Although changes in various transporters in the liver and kidney in cholestasis have been elucidated, little is known about how effectively the elimination of these compounds is compensated for by urinary excretion. PMID- 15127277 TI - Combined devascularization and proximal splenorenal shunt: is this a better option than either procedure alone? AB - BACKGROUND/PURPOSE: We aimed to determine the rationality of pericardial devascularization (PCDV) plus proximal splenorenal shunt (PSRS) for cirrhotic patients with portal hypertension with variceal bleeding, using a duplex sonography study of the effects of the different surgical procedures (PCDV, PSRS, and PCDV + PSRS) on the hemodynamics of the portal system. METHODS: Ninety-nine patients with cirrhotic portal hypertension and a history of bleeding esophageal varices were studied. These patients were divided into three groups (PSRS group, PCDV group, and PCDV + PSRS group). The hemodynamic parameters of the portal systems of all patients were measured by Doppler color-flow imaging perioperatively. RESULTS: In the PSRS group, the postoperative portal venous flow (PVF) and free portal pressure (FPP) decreased by 57 +/- 9% and 52 +/- 5%, respectively (P < 0.01). In the PCDV group, the postoperative PVF lessened by 8 +/- 5% (P > 0.05), and the postoperative FPP was reduced by 19 +/- 7% (P < 0.05). In the PCDV + PSRS group, the postoperative PVF and FPP were lowered by 36 +/- 8% and 34 +/- 10%, respectively (P < 0.05). The postoperative decreases of PVF and FPP in the PCDV + PSRS group were between those of the PSRS and PCDV groups. The differences among these groups were statistically significant (P < 0.05). CONCLUSIONS: Combined devascularization and splenorenal shunt (PCDV + PSRS) significantly decreases portal venous flow and portal pressure, as well as maintaining hepatopedal flow, thus entailing fewer complications compared to either PCDV or PSRS. We aimed to determine the rationality of pericardial devascularization (PCDV) plus proximal splenorenal shunt (PSRS) for cirrhotic patients with portal hypertension with variceal bleeding, using a duplex sonography study of the effects of the different surgical procedures (PCDV, PSRS, and PCDV + PSRS) on the hemodynamics of the portal system. PMID- 15127278 TI - Is drip infusion cholecystocholangiography (DIC) an acceptable modality at cholecystectomy for cholecystolithiasis, considering the frequency of bile duct maljunction and intraoperative bile duct injury? AB - BACKGROUND/PURPOSE: Knowledge of the configuration of the extrahepatic bile duct is indispensable to avoid bile duct injury during cholecystectomy. Various methods of examining the biliary tract have been developed; however, the most appropriate preoperative diagnostic modality at cholecystectomy for cholecystolithiasis has not yet been reported. Considering the frequency of bile duct maljunction (BDM) and operative bile duct injury, in addition to the cost and invasiveness of the various examination methods, we evaluated the usefulness of drip infusion cholecystocholangiography (DIC) as the optimal method of examination at cholecystectomy for cholecystolithiasis. METHODS: Preoperative diagnostic accuracy of BDM was analyzed in relation to operative bile duct injury using 469 patients with benign biliary diseases who was diagnosed with DIC and endoscopic retrograde cholecystography and underwent cholecystectomy. BDM was classified according to Hisatsugu criteria. RESULTS: Out of 469 consecutive patients who underwent a cholecystectomy for biliary stones between January 1, 1995, and September 30, 1998, at Ohta Nishinouchi General Hospital, 21 (4.48%) had a cystic duct maljunction (CDM) and 12 (2.56%) had an aberrant bile duct (ABD). The most common variants were types C and D for CDM, and types II and III for ABD, according to Hisatsugu's classification. Fourteen patients (42.4%) were diagnosed before the surgery; 13 of them received preoperative endoscopic retrograde cholangiography (ERC), and the remaining patient underwent preoperative drip infusion cholecystocholangiography (DIC). Nineteen patients could not be correctly diagnosed based on their preoperative examinations, but were diagnosed during surgery. Operative bile duct injury occurred in 1 patient (0.2%) whose maljunction could not be diagnosed before the operation. CONCLUSIONS: Taking into account the medical cost and invasiveness, and the frequency of BDM and related bile duct injuries, we conclude that DIC is an acceptable preoperative diagnostic modality to employ at cholecystectomy for cholecystolithiasis. Knowledge of the configuration of the extrahepatic bile duct is indispensable to avoid bile duct injury during cholecystectomy. Various methods of examining the biliary tract have been developed; however, the most appropriate preoperative diagnostic modality at cholecystectomy for cholecystolithiasis has not yet been reported. Considering the frequency of bile duct maljunction (BDM) and operative bile duct injury, in addition to the cost and invasiveness of the various examination methods, we evaluated the usefulness of drip infusion cholecystocholangiography (DIC) as the optimal method of examination at cholecystectomy for cholecystolithiasis. PMID- 15127279 TI - Clear cell carcinoma of the pancreas: an adenocarcinoma with unusual phenotype of duct cell origin. AB - Although clear cell carcinoma has been found in various organs, only six cases have been reported in the pancreas. Moreover, the histogenesis of clear cell carcinoma of the pancreas remains controversial. We report a case of clear cell carcinoma of the pancreas in a 61-year-old woman, with an unusual pheno- or genotype detected by histochemical, immunohistochemical, and K- ras oncogene analyses. Histologically, the pancreatic tumor was predominantly composed of clear cell nests with scanty fibrous stroma and scattered duct-like structures. Neither clear cell nor duct-like components of the tumor showed mucin production. Immunohistochemical analysis of neoplastic cells showed a positive reaction to antibodies against cytokeratins 8 and 19, carbohydrate antigen 19-9, and alpha-1 antitrypsin, and showed no reaction to antibodies against carcinoembryonic antigen, neuroendocrine markers, trypsin, amylase, and HMB45. K- ras analysis revealed no mutation at codon 12 in either clear cell or duct-like components. The patient has had no recurrence as yet. The pancreatic carcinoma in our patient may be of duct cell origin, but the results of histochemical, immunohistochemical, and gene analyses and patient's outcome were unusual compared with those of previous cases. PMID- 15127280 TI - Ruptured pancreaticoduodenal artery aneurysm treated by superselective transcatheter arterial embolization and preserving vascularity of pancreaticoduodenal arcades. AB - We report a case of a ruptured aneurysm in the anterior superior pancreaticoduodenal artery (PDA) with hypovolemic shock managed successfully by superselective transcatheter arterial embolization of the aneurysm. A 75-year-old male presented to our hospital with hematemesis and melena. On admission, he was in shock. Angiography showed an aneurysm about 1 cm in diameter in the anterior superior PDA. However, extravasation of contrast medium was not seen owing to hypovolemic shock. A catheter was inserted into the aneurysm, and superselective microcoil embolization of the PDA aneurysm was successfully achieved. After the microcoil was inserted into the aneurysm itself, it was observed that duodenal vascularity and pancreaticoduodenal arcades were preserved and aneurysm was not present. There was no complication such as necrosis or abscess formation in the pancreas. The patient recovered and is doing well after 18 months of follow-up. Superselective transcatheter arterial embolization should be considered as the initial treatment of choice for all peripancreatic aneurysms. PMID- 15127281 TI - Repeated resection for extrahepatic recurrences after hepatectomy for ruptured hepatocellular carcinoma. AB - In March 1999, a 54-year-old man with chronic hepatitis C was referred to our hospital because of ruptured hepatocellular carcinoma (HCC) located in Couinaud's segments 4 and 8. He underwent central bisegmentectomy of the liver with partial resection of the diaphragm. After the first surgery, extrahepatic metastases were found on different occasions in the abdominal wall, thoracic cavity, and greater omentum and were all surgically resected. In February 2001, the serum protein induced by vitamin K absence or antagonist-II (PIVKA-II) level increased markedly to 19,000 mAU/l. Magnetic resonance imaging showed a massive right subphrenic tumor with invasion to the right diaphragm and posterior segment of the liver. The patient underwent en bloc resection of the tumor, diaphragm, posterior segment of the liver, and right lower pulmonary lobe. After the surgery, the PIVKA-II level rapidly decreased, and it has remained within the normal range to date. Two years after the last surgery, the patient is doing well without any extrahepatic recurrence, although small intrahepatic recurrences have been completely treated by radiofrequency ablation and transcatheter arterial chemoembolization. Ruptured HCC often exacerbates the risk of peritoneal dissemination and is usually difficult to completely resect. This is an extremely rare case of a patient who successfully underwent five repeated resections for extrahepatic recurrences after hepatectomy for ruptured HCC. PMID- 15127282 TI - Effects of brief pain education on hospitalized cancer patients with moderate to severe pain. AB - The purpose of this randomized controlled study was to assess the effects of a structured pain education program on the pain experience of hospitalized cancer patients. Eligible cancer pain patients were randomly assigned to either an experimental group (receiving pain education 10-15 min per day for 5 days, n = 15) or a standard care control group (n = 15). The effects of the intervention on six pain-related variables were evaluated using three instruments. Pain intensity, pain interference with daily life, negative beliefs about opioids, beliefs about endurance of pain, pain catastrophizing (an individual's tendency to focus on and exaggerate the threat value of painful stimuli and negatively evaluate his or her own ability to deal with pain), and sense of control over pain were evaluated by the Brief Pain Inventory-Short Form Taiwanese version (BPI T), Pain and Opioid Analgesic Beliefs Scale-Cancer (POABS-CA), and the Catastrophizing subscale and the sense of control over pain measure from the Coping Strategies Questionnaire (CSQ). The results indicated that, after completing treatment, patients who had received structured pain education had significantly less pain intensity on average, negative pain beliefs regarding opioids, pain endurance beliefs, and pain catastrophizing than patients in the control group. In addition, patients in the pain education group showed a significant increase in their sense of control over pain. These preliminary results strongly suggest that structured pain education can effectively improve the pain experience of hospitalized cancer patients and should be further implemented clinically. PMID- 15127283 TI - Mild antenatal hydronephrosis: management controversies. PMID- 15127284 TI - High factor VIII levels during attacks of familial Mediterranean fever. PMID- 15127285 TI - Spatial associations among algae affect host use in a herbivorous marine amphipod. AB - Host plant choice by herbivores may be constrained by the spatial arrangement of plants with selection of poor quality hosts being more likely if preferred hosts are locally rare. I tested whether the presence of a herbivorous marine amphipod, Peramphithoe parmerong, on a poor quality host could be explained by the relative abundance and spatial arrangement of alternative brown algal hosts. Amphipods strongly preferred the high quality Sargassum linearifolium over the poor quality Padina crassa in laboratory assays with a higher tendency to select Padina crassa when it was relatively more abundant. In the field, however, adults colonised both hosts to equal densities. Rates of colonisation were independent of host quality but strongly affected by the identity of neighbouring hosts with those algae in close proximity to S. linearifolium receiving far greater densities of amphipods than those surrounded by Padina crassa. There was no evidence that the presence of adult Peramphithoe parmerong on Padina crassa was due to local scarcity of S. linearifolium. In contrast, the distribution and behaviour of juveniles were predictable from differences in food quality, with most inhabiting the high quality S. linearifolium and few emigrating from this host. The success of a simple adaptive model based on food quality to predict juvenile, but not adult, behaviour indicates that the relative importance of food quality varies with age, and that other factors are important determinants of adult movements among hosts. PMID- 15127286 TI - The influence of swimming demand on phenotypic plasticity and morphological integration: a comparison of two polymorphic charr species. AB - In northern freshwater lakes, several fish species have populations composed of discrete morphs, usually involving a divergence between benthic and limnetic morphs. Although it has been suggested that swimming demand plays an important role in morphological differentiation, thus influencing habitat selection, it is unclear how it affects reaction norms, patterns in character correlation, and levels of morphological integration. We examined whether swimming demand could induce morphological plasticity in the directions expected under divergent habitat selection, and evaluated its influence on the morphological integration in Arctic charr ( Salvelinus alpinus) and brook charr ( S. fontinalis), two congeneric species exhibiting conspicuous and subtle resource polymorphism, respectively. We found that changes in morphology were induced by differential swimming demands in both species. The length of the pectoral fin was the character that responded most strongly according to the predicted morphological expectations under divergent habitat selection. High levels of morphological plasticity, relatively low levels of integration, and differences found in the morphological correlation structure among water velocity treatments suggest that constraints on morphological change are unlikely in either species, thus allowing great potential for phenotypic flexibility in both species. The magnitude of character integration, however, was larger for Arctic charr than for brook charr. This latter result is discussed in the light of the differences in the level of polymorphism between the two species in the wild. The results of the present study indicate that swimming demand alone may not be sufficient to generate the polymorphism encountered in nature. Given that both diet and swimming demands can induce morphological changes, it would be important to conduct experiments targeting the interaction between the morphological modules related to trophic and swimming demands. PMID- 15127287 TI - Hatching asynchrony, survival, and the fitness of alternative adult morphs in Ambystoma talpoideum. AB - The mole salamander, Ambystoma talpoideum, exhibits both aquatic (gilled) and terrestrial (metamorphosed) adult morphologies. Previous studies have shown the existence of body-size advantages associated with the terrestrial morph in A. talpoideum and other polymorphic salamanders (e.g., A. tigrinum). However, aquatic adult A. talpoideum mature at a younger age and often breed earlier than terrestrial adults. We tested the hypothesis that early maturation and reproduction in aquatic adults increase fitness (irrespective of body size). We reared larval A. talpoideum in mesocosms and varied the timing of hatching, with early-hatching larvae representing the offspring from early-breeding aquatic adults, and late-hatching larvae representing the offspring of later-breeding terrestrial adults. Our results demonstrate significantly higher survival rates among early-hatchlings relative to late-hatching conspecifics, supporting the hypothesis that early reproduction may be an important mechanism mediating the polymorphism in A. talpoideum. We discuss our results within the context of size based models of the fitness of alternative life-cycles. PMID- 15127288 TI - Nestin, a neuroectodermal stem cell marker molecule, is expressed in Leydig cells of the human testis and in some specific cell types from human testicular tumours. AB - The intermediate filament protein nestin is predominantly expressed in some stem/progenitor cells and appears to be a useful molecular tool to characterise tumours originating from precursor cells of neuroectodermal and mesenchymal lineages. Leydig cells originate in the adult testis by differentiation from stem cells and express a variety of neural and neuroendocrine markers. The possible expression of the neural stem cell marker nestin in Leydig cells and testicular tumour cells was determined by analysing the patterns of nestin expression in normal and pathological human testes by Western blot and immunohistochemical methods. In normal testis, nestin was found in some vascular endothelial cells, a subset of peritubular spindle-shaped cells and some Leydig cells; spermatogenic and Sertoli cells were unstained. In normal Leydig cells, nestin was distributed in the perinuclear cytoplasm and accumulated in the crystalloids of Reinke with ageing. In non-tumour pathologies (cryptorchidism, impaired spermatogenesis), the seminiferous tubules were immunonegative, whereas hyperplastic Leydig cells showed cytoplasmic immunolabelling. In testicular malignancies, nestin was localised in the Sertoli cells of the seminiferous tubules affected with intratubular germ cell neoplasia, in the hyperplastic Leydig cells associated with these tumours and in some components (mesenchymal and neuroepithelial cells) of teratomas; spermatocytic and non-spermatocytic seminomas were unstained. Some vascular endothelial cells were immunolabelled in all tumour samples. Thus, nestin is expressed in a population of normal and hyperplastic Leydig cells and in Sertoli cells in the presence of intratubular germ-cell neoplasia. Nestin may be a good marker for identifying components of testicular teratomas. PMID- 15127289 TI - Tissue specific expression and sequence analysis of a stress responsive gene Bre in adult golden hamster (Mesocricetus auratus). AB - Bre (brain and reproductive organ-expressed) is a new and putative stress modulating gene of yet unknown function. BRE has previously been shown to interact with type 1 tumor necrosis factor receptor (TNFR1) and modulate the action of TNF. Apart from the brain and reproductive organs, Bre and BRE are highly expressed in steroid producing tissues such as the adrenal gland. Here we report for the first time the cloning of the Bre gene from golden hamster, a model organism extremely valuable for reproduction and steroid research, and examination of its tissue specific expression. Sequence analysis demonstrated that the peptide sequence of BRE in hamster shares approximately 99% homology with those of human, monkey and mouse. The hamster Bre gene transcribed an approximately 1.8-kb mRNA which translated a 44-kDa protein. Bre was strongly expressed in neurons and luminal epithelia of urogenital, digestive and respiratory organs. Bre was also detected in lymphoid tissues and endocrine glands. Immunohistochemistry demonstrated a similar protein expression pattern. Exceptions to this included the adrenal gland, where a high level of Bre was accompanied by weak immunoreactivity; as well as the oocytes and islets of Langerhans, where BRE protein but not the mRNA was localized. These data indicated that Bre gene products were expressed in a wide variety of tissues other than the brain and reproductive organs, as was originally described. Based on our findings, we propose that Bre is a housekeeping gene in tissues that are constantly subjected to environmental hazards such as luminal epithelia. Our results further support the proposed role for BRE in endocrine and immune functions. PMID- 15127291 TI - Somatic mutations of CASP3 gene in human cancers. AB - Failure of apoptosis is one of the hallmarks of cancer. As an execution-phase caspase, caspase-3 plays a crucial role during apoptosis. To explore the possibility that the genetic alterations of CASP3, which encodes caspase-3, might be involved in the development of human tumors, we analyzed the entire coding region and all splice sites of human CASP3 gene for the detection of somatic mutations in a series of 944 human tumors, including 165 stomach carcinomas, 95 colon carcinomas, 76 breast carcinomas, 80 hepatocellular carcinomas, 181 non small cell lung cancers, 45 acute leukemias, 28 multiple myelomas, 12 medulloblastomas, 15 Wilms' tumors, 12 renal cell carcinomas, 40 esophagus carcinomas, 33 urinary bladder carcinomas, 33 laryngeal carcinomas, and 129 non Hodgkin lymphomas. Overall, we detected 14 somatic mutations of the CASP3 gene, including six missense and four silent mutations, two mutations in the introns, one mutation in the 5'-untranslated region, and one mutation in the 3' untranslated region. The mutations were observed in four of 98 colon carcinomas (4.1%), four of 181 non-small cell lung cancers (2.2%), two of 129 non-Hodgkin lymphomas (1.6%), two of 165 stomach carcinomas (1.2%), one of 80 hepatocellular carcinomas (1.3%), and one of 28 multiple myelomas (3.6%). This is the first report on CASP3 gene mutations in human tumors; these data indicate that the CASP3 gene is occasionally mutated in human tumors. PMID- 15127290 TI - Lipoprotein lipase gene is in linkage with blood pressure phenotypes in Chinese pedigrees. AB - To elucidate the mechanism of lipid metabolism in the genesis of essential hypertension (EH), we linked blood pressure (BP) phenotypes with the lipoprotein lipase (LPL) gene. Variance component and sib-pair linkage models were used to test the relationship of the polymorphisms in the LPL gene region and EH in 148 Chinese hypertensive families. Linkage evidence with systolic BP (SBP) and diastolic BP (DBP) was observed in a total population of 148 pedigrees with seven flanking microsatellite markers of the LPL gene, with a maximum two-point LOD score of 2.68 and a maximum multipoint LOD score (MLS) of 2.37 for SBP and a maximum MLS of 1.54 for DBP. Suggestive linkage results around this region were also obtained in northern and southern subsets by geographic distribution. In addition, quantitative-transmission/disequilibrium-test analyses showed that there was linkage between DBP and two single nucleotide polymorphisms in the LPL gene. This is the first report of linkage between LPL gene and DBP in the Chinese population. The LPL gene itself might explain our results or the LPL gene region might harbor some genes to explain the observed results to some degree and might contribute to the variation of BP in the Chinese population. PMID- 15127292 TI - The influence of parasitism on the pharmacokinetics of moxidectin in lambs. AB - Most pharmacokinetic studies on anthelmintic drugs have been performed on non parasitized animals. However, it seems likely that the parasite burden could influence the deposition of such drugs. The pharmacokinetics of moxidectin administered orally and by subcutaneous injection was compared in lambs exposed to nematode infection and in parasite naive lambs. Plasma samples were analyzed for moxidectin over 40 days post-treatment. The main pharmacokinetic parameters calculated demonstrated a significant change in drug deposition in infected lambs when compared to controls. The area under the plasma concentration-time curve was decreased 54% and 46% by infection in the subcutaneous and oral groups, respectively. There was also a major decrease in the mean residence time in parasitized lambs. In parallel, the clearance of the drug was increased by infection. Thus, parasite infection dramatically influences the disposition of moxidectin in lambs. These results may contribute to determining a therapeutic strategy adapted to heavily infested animals. PMID- 15127293 TI - Purification, characterization and kinetic properties of the Taenia solium glutathione S-transferase isoform 26.5 kDa. AB - Glutathione S-transferases are major phase II detoxification enzymes. Taenia solium, a parasite of humans and pigs, is exposed to toxic products. The aim of this work was to purify and characterize a T. solium glutathione S-transferase isoform of 26.5 kDa (SGST26.5) in order to obtain its kinetic parameters. Homogeneous SGST26.5 was obtained by a simple purification procedure. SGST26.5 showed a p I of 7.07, and a native Mr of 60 kDa with 26.5 kDa subunits. The optimum activity for SGST26.5 was found at pH 6.5-7.0 in the range 10-42 degrees C. SGST26.5 had a specific enzyme activity of 78, 7.1, 6.6, and 0.7 microM min( 1) mg(-1) with CDNB, 1,2-dichloro-4-nitrobenzene, 2,4-hexadienal and trans-2 nonenal as substrates, respectively. It also had a kcat/ K(mCDNB)=2.15 x 10(3) M( 1 )s(-1), kcat/ KmGSH)=4.5 x 10(3) M(-1 )s(-1) and Vmax for GSH and CDNB=74 and 77 microM min(-1) mg(-1), respectively. SGST26.5 was inhibited in a noncompetitive form by cibacron blue, bromosulfophthalein and triphenyltin chloride. Inhibition studies as a function of inhibitor concentration show that the enzyme is a homodimer. Bireactant system analysis show that it follows an ordered sequential mechanism. PMID- 15127294 TI - Morphology of the rediae of Echinostoma paraensei (Trematoda: Echinostomatidae) from its intermediate host Lymnaea columella (Mollusca, Gastropoda). AB - The morphology of the rediae of Echinostoma paraensei obtained from Lymnaea columella was studied using light, scanning and transmission electron microscopy. The measurements of the mature rediae differ from those described originally, and the taxonomic importance of the ambulatory buds and papilliform process is discussed. Uniciliated papillae were observed in the mouth region. The birth papilla is a bulb-like structure, well defined at the anterior end of the body of the rediae, which opens through a split. There are no microvilli in the tegument surface of the larvae, but numerous tegumental folds, varying according to the contraction of the body of the rediae. The outer syncytial layer is located on a thick basal lamina below which the circular and the longitudinal muscle fiber layers are located. PMID- 15127295 TI - Echinococcus granulosus strain typing in Bulgaria: the G1 genotype is predominant in intermediate and definitive wild hosts. AB - Addressing the genetic variability in Echinococcus granulosus is epidemiologically important, as strain characteristics may influence the local transmission patterns of zoonotic cystic echinococcosis. To classify the genotype(s) present in intermediate (pig, cattle and sheep) and definitive (jackal and wolf) hosts in Bulgaria, a DNA-based approach was used to assess parasite protoscoleces or strobiles. Genes corresponding to coding and non-coding regions of the nuclear and mitochondrial genome ( ND-1, HBX, Act II, AgB-1) were amplified by PCR and subsequently sequenced. The sequences resolved were all found to be identical to those published for the common sheep strain of E. granulosus, indicating that the G1 genotype is predominant in Bulgaria. One microvariant for ND-1 was found in the pig isolates; however no epidemiological significance was attributed to this finding. PMID- 15127296 TI - A trypanosome of Atlantic cod, Gadus morhua L., transmitted by the marine leech Calliobdella nodulifera (Malm, 1863) (Piscicolidae). AB - The fish leech, Calliobdella nodulifera, transmitted a trypanosome while feeding on laboratory-reared cod ( Gadus morhua). The flagellate body length increased from 43 microm 17 days post-infection (d.p.i.) to 57 microm 113 d.p.i. Characteristic features are cell striation, a nearly central nucleus and a short (3.5 microm) distance from the kinetoplast to the posterior end (PK). Following growth, the trypanosomes became increasingly slender, with fewer striae and a shorter flagellum, while the PK was unaffected. The trypanosomes differ morphologically from Trypanosoma murmanensis transmitted by the leech Johanssonia arctica. PMID- 15127297 TI - Molecular characterization of a Brugia malayi transglutaminase. AB - Prior studies have demonstrated that transglutaminase (TGase) from the human filarial parasite Brugia malayi is critical for the growth and development of the larval stages. In this report, we describe the cloning and partial characterization of a cDNA encoding the B. malayi TGase (BmTGase). Using RT-PCR and RACE-PCR, the cDNA was amplified from adult worm mRNA. BmTGase is 1,881 bp long and codes for a protein with a predicted molecular mass of 54 kDa. Amino acid sequence analysis of BmTGase revealed significant homology to the protein disulfide isomerase (PDI), particularly, to the PDI-related protein ERp60, a PDI isoform found in the lumen of endoplasmic reticulum. The activity of recombinant B. malayi TGase enzyme (rBmTG) was found to be calcium-dependent and could be inhibited by EDTA. ELISA studies showed that approximately 88% of 48 sera from healthy Indian patients living in a bancroftian filariasis endemic area were reactive with rBmTG. In contrast, only 33% of sera from patients with clinical filariasis were reactive to rBmTG. Non-endemic sera were uniformly non-reactive. Additional studies are needed to elucidate the role, if any, of B. malayi TGase in protective immunity to filariasis. PMID- 15127298 TI - Seasonal dynamics of Posthodiplostomum cuticola (Digenea, Diplostomatidae) metacercariae and parasite-enhanced growth of juvenile host fish. AB - The seasonal dynamics of Posthodiplostomum cuticola metacercariae in 0+ juvenile fish, Rutilus rutilus, Scardinius erythrophthalmus and Abramis bjoerkna, was studied on the floodplain of the Dyje River, Czech Republic. Prevalence and mean abundance of P. cuticola were significantly higher in R. rutilus than in S. erythrophthalmus or A. bjoerkna. A seasonal pattern of parasite infection with maximum values in autumn was evident in all three species. No effect of overwintering on the P. cuticola infection was detected. Parasite-induced growth was found for all three fish species investigated; the fish standard length and body weight of parasitized individuals were significantly higher than those of unparasitized fish from July to October. In April, no difference was found. The maximum enhanced growth of parasitized fish was found in months with low zooplankton densities, while the difference was lower when food was abundant. PMID- 15127299 TI - Co-expression of tyrosine hydroxylase, dopamine beta-hydroxylase and neuropeptide Y in the sympathetic neurons projecting to the submandibular gland in the sheep. AB - Sympathetic neurons projecting to the ovine submandibular gland (SMG) from the superior cervical ganglion (SCG) and middle cervical ganglion (MCG) were identified using retrograde tracing with fluorescent dye (Fast Blue). Antibodies to tyrosine hydroxylase (TH), dopamine beta-hydroxylase (DbetaH) and neuropeptide Y (NPY) were used to determine the immunochemical characteristics of SMG innervating sympathetic neurons. Immunohistochemistry combined with the retrograde tracing revealed that the population of SMG-projecting neurons consist of four distinct sub-populations, but taking into account their possible different physiological properties only three major sub-populations can be distinguished. The vast majority of neurons in both ganglia are noradrenergic in nature (co-express TH and DbetaH). All examined TH-immunoreactive (IR) neurons also show immunoreactivity to DbetaH. Sub-population of noradrenergic neurons can be divided into NPY-IR and non-NPY-IR. Noradrenergic neurons expressing NPY may act as vasoconstrictors. The second sub-population of SMG projecting neurons in the ganglia studied consists of non-noradrenergic neurons (containing NPY, but not TH). It is known that these kinds of neurons may play a vasodilatory role. In both examined ganglia the third sub-population consists of non-TH-IR and non-NPY IR neurons of unknown physiological function. Since no DbetaH immunoreactivity was found in any of TH- neurons these nerve cells can also be regarded as non noradrenergic. It is possible that some neurons of the second as well as the third sub-population are cholinergic and some of them are non-noradrenergic/non cholinergic in character. PMID- 15127301 TI - Perturbations of malate accumulation and the endogenous rhythms of gas exchange in the Crassulacean acid metabolism plant Kalanchoe daigremontiana: testing the tonoplast-as-oscillator model. AB - In continuous light, leaves of the Crassulacean acid metabolism (CAM) plant Kalanchoe daigremontiana Hamet et Perrier exhibit a circadian rhythm of CO2 uptake, stomatal conductance and leaf-internal CO2 pressure. According to a current quantitative model of CAM, the pacemaking mechanism involves periodic turgor-related tension and relaxation of the tonoplast, which determines the direction of the net flux of malate between the vacuole and the cytoplasm. Cytoplasmic malate, in turn, through its inhibitory effect on phospho enolpyruvate carboxylase, controls the rate of CO2 uptake. According to this mechanism, when the accumulation of malate is disrupted by removing CO2 from the ambient air, the induction of a phase delay with respect to an unperturbed control plant is expected. First, using the mathematical model, such phase delays were observed in numerical simulations of three scenarios of CO2 removal: (i) starting at a trough of CO2 uptake, lasting for about half a cycle (ca. 12 h in vivo); (ii) with the identical starting phase, but lasting for 1.5 cycles (ca. 36 h); and (iii) starting while CO2 increases, lasting for half a cycle again. Applying the same protocols to leaves of K. daigremontiana in vivo did not induce the predicted phase shifts, i.e. after the end of the CO2 removal the perturbed rhythm adopted nearly the same phase as that of the control plant. Second, when leaves were exposed to a nitrogen atmosphere for three nights prior to onset of continuous light to prevent malate accumulation, a small, 4-h phase advance was observed instead of a delay, again contrary to the model-based expectations. Hence, vacuolar malic acid accumulation is ruled out as the central pacemaking process. This observation is in line with our earlier suggestion [T.P. Wyka, U. Luttge (2003) J Exp Bot 54:1471-1479] that in extended continuous light, CO2 uptake switches gradually from a CAM-like to a C3-like mechanism, with oscillations of the two CO2 uptake systems being tightly coordinated. It appears that the circadian rhythm of gas exchange in this CAM plant emerges from one or several devices that are capable of generating temporal information in a robust manner, i.e. they are protected from even severe metabolic perturbations. PMID- 15127302 TI - PGE2 stimulates Cl- secretion in murine M-1 cortical collecting duct cells in an autocrine manner. AB - Prostaglandin E2 (PGE2) is thought to be an important modulator of renal ion and water transport, but its effects remain complex and incompletely understood. Here we examined the effects of PGE2 on transepithelial ion transport of M-1 mouse cortical collecting duct cells using short-circuit current (ISC) measurements. Basolateral addition of PGE2 (1 microM) produced a transient peak increase in ISC of 6.3+/-0.8 microA cm(-2) (n=11), followed by a sustained plateau. The PGE2 evoked response was preserved in the presence of 100 micro M apical amiloride with an average peak increase of 10.6+/-1.0 microA cm(-2) (n=23). However, it was greatly diminished in both the presence of apical diphenylamine-2-carboxylic acid (DPC, 1 mM) and the absence of extracellular Cl-, indicating that Cl- secretion had been stimulated. Basolateral PGE2 induced a concentration dependent response, with an EC50 of about 8 nM. Apical addition of PGE2 elicited an ISC response similar to that observed with basolateral PGE2. Furthermore, apical exposure to arachidonic acid (AA) produced a similar increase in ISC, which could be prevented by the cyclooxygenase inhibitor indomethacin, while AA failed to exert an additional effect in the presence of PGE2. Using RT-PCR, we confirmed the expression of the PGE2 (EP) receptor subtypes EP1, EP3 and EP4 but not of EP2 in cultured M-1 CCD cells. We conclude that M-1 cells express functional cyclooxygenase activity and can generate PGE2 which acts in an autocrine manner, causing Cl- secretion. PMID- 15127304 TI - Ventricular tapping seems to have no influence on S-100B and NSE serum concentrations. AB - Serum markers, e.show $132#g., the protein S-100B and neuron-specific enolase (NSE), are recognized to give additional information about the extension and prognosis of brain damage. In some of these patients it is necessary to insert a ventricular drain. Whether the cannulation of the ventricle falsifies the serum concentrations of these markers is unknown. The aim of this study was to get further information in this field. In this prospective study we included 19 patients. All patients underwent ventricular tapping and insertion of a ventricular drain. Serum samples for estimation of S-100B and NSE were collected before, directly after and 6 h after insertion. In addition we investigated the cerebrospinal fluid (CSF) directly after and 6 h after insertion. All patients but one showed no significantly increased S-100B or NSE serum concentration after insertion of the drainage. The concentrations in the CSF were significantly higher. One patient showed increasing concentrations of the markers in all samples reflecting ongoing brain damage. The serum values of S-100B and NSE seem not to be falsified by insertion of a ventricular drain. Therefore the prognostic value of these serum markers seems to be preserved despite the surgical manipulation. PMID- 15127305 TI - Molecular modeling of B-DNA site recognition by Ru intercalators: molecular shape selection. AB - In this work, molecular modeling methods have been applied to the interaction characterization of polypyridyl transitional-metal complexes with the oligonucleotide (B-DNA fragment). In order to explore the factors governing the groove recognition and intercalative depth, we establish a simple and practical docking method (step-by-step docking operation) to obtain potential curves while making complexes inset into B-DNA along an assigned path. Energy values in the potential curve are obtained from energy minimization of binding geometries. Modeling results clearly show that the optimum binding conformation corresponding to the global minimum in the potential curve for each complex is found to correlate well with the experimental results. Our results also confirm that minor changes of the ligand structure can lead to profound influences on binding geometries, so the molecular shape of the complexes is a predominant factor in governing the binding mode. Moreover, we find that the vdW force and "water molecular effect" are strongly associated with molecular-shape selection in our model. These results complement and extend the knowledge of the nature of these complexes binding to B-DNA. PMID- 15127306 TI - Anaerobic, alkaliphilic, saccharolytic bacterium Alkalibacter saccharofermentans gen. nov., sp. nov. from a soda lake in the Transbaikal region of Russia. AB - Three strains of new obligately anaerobic alkaliphilic bacteria have been isolated as a saccharolytic component from the cellulolytic community of alkaline Lake Nizhnee Beloe (Transbaikal region, Russia), a lake with low salt concentration. DNA analysis of these strains showed an interspecies level of DNA similarity of 96-100%. Strain Z-79820 was selected for further investigations. Cells were Gram-positive, asporogenous, nonmotile short rods with pointed ends. The strain was a true alkaliphile: growth occurred from pH 7.2 to 10.2 with the optimum at pH 9.0. Strain Z-79820 was halotolerant and could grow in medium with up to 10% (w/v) NaCl, with the optimum between 0 and 4% NaCl. The new isolate obligately depended on Na+ ions in the form of carbonates or chlorides. Total Na+ content needed for optimal growth was 0.46 M Na+, with a wide range from 0.023 0.9 M Na+ at which growth also occurred. The isolate was a mesophile and grew at temperatures from 6 to 50 degrees C (slow growth at 6 and 15 degrees C) with an optimum at 35 degrees C. The organotrophic organism fermented ribose, xylose, glucose, mannose, fructose, sucrose, mannitol, and peptone. The products of glucose fermentation were acetate, ethanol, formate, H2, and CO2. Yeast extract was required for some anabolic needs. The DNA G+C content of the type strain Z 79820 was 42.1 mol%. The new bacterium fell into the 16S rRNA gene cluster XV of the Gram-positive bacteria with low G+C content, where it formed an individual branch. Based on its growth characteristics and genotype traits, we propose the new genus and species named Alkalibacter saccharofermentans with the type strain Z-79820 (=DSM14828), Uniqem-218 (Institute Microbiology, RAS; http://inmi.da.ru). PMID- 15127307 TI - Cerebrospinal fluid concentrations of idebenone in Friedreich ataxia patients. AB - We studied plasma and cerebrospinal fluid (CSF) concentrations of idebenone in five Friedreich ataxia patients on treatment with this antioxidant, and plasma and CSF ubiquinone-10 (Q (10)) concentrations in 15 controls. CSF idebenone concentrations were below the detection limit in 3 Friedreich ataxia patients and no association could be demonstrated between plasma and CSF idebenone values. Q (10) CSF concentrations (median: 2.25 nmol/L) were approximately 300 times lower than those of plasma (median: 0.77 micro mol/L). No correlation was observed between plasma and CSF Q (10) concentrations. A significantly positive correlation was observed between CSF total protein values (range 8.1 - 107.5 mg/dL; median: 29.5) and CSF Q (10) concentrations (Spearman test: r = 0.664; p = 0.01). Our findings suggest that less idebenone is distributed to the brain than to other tissues, although CSF does not appear to be an appropriate material for treatment monitoring of idebenone and other quinoid compounds. PMID- 15127308 TI - Comorbidity headache and epilepsy in childhood. AB - RATIONALE: Epilepsy and headache are both frequent in childhood. Because seizures are frequently a frightening event, other medical conditions--including headache- are often neglected not only by the patient, but also by the physician. The objective of this study was to verify the comorbidity between headache and epilepsy in childhood. METHODS: This was a prospective study conducted at the pediatric epilepsy clinic of our university hospital. Fifty children with epilepsy and ability to describe their symptoms, between 5 and 18 years old, were interviewed according to a semi-structured questionnaire. The headache was classified according to the International Headache Society. The frequency of headache was compared with the findings of a control group composed by children without epilepsy, siblings of children with epilepsy. RESULTS: Fifty children were evaluated, 29 boys, mean age 11 years. Twenty-three (46 %) patients presented with headache, as opposed to only 1 (2.5 %) in the control group ( p < 0.01). Ten (43.5 %) had migraine, 4 (17.4 %) had tension type headache and in 9 (39.1 %) the type of headache could not be established. In 9/23 (39 %) a temporal relationship between headache and epilepsy was present, 6 postictal and 3 preictal. There was no difference in gender, age, type of seizure and family history of headache in the groups of patients with or without headache. However, most patients with headache were older than 10 years (54.5 %) and had idiopathic epilepsy (65.2 %; p < 0.01). The headache usually started in the same year or after the diagnosis of epilepsy (95 %; p < 0.01). CONCLUSION: Headache and epilepsy are a common comorbidity in childhood, and occur mostly in children older than 10 years with idiopathic epilepsy. The headache usually starts in the same year or after the diagnosis of epilepsy. PMID- 15127309 TI - Collagen VI status and clinical severity in Ullrich congenital muscular dystrophy: phenotype analysis of 11 families linked to the COL6 loci. AB - Ullrich's congenital muscular dystrophy (UCMD) is an autosomal recessive myopathy characterised by neonatal muscle weakness, proximal joint contractures and distal hyperlaxity. Mutations in the COL6A1, COL6A2 (21 q22.3) and COL6A3 (2 q37) genes, encoding the alpha 1, alpha 2 and alpha 3 chains of collagen VI, respectively, have been recently identified as responsible for UCMD in a total of 9 families. We investigated in detail the clinical and morphological phenotype of 15 UCMD patients from 11 consanguineous families showing potential linkage either to 21 q22.3 (6 families) or to 2 q37 (5 families). Collagen VI deficiency was confirmed on muscle biopsies or skin fibroblasts in 8 families. Although all patients shared a common phenotype, a great variability in severity was observed. Collagen VI deficiency in muscle or cultured fibroblasts was complete in the severe cases and partial in the milder ones, which suggests a correlation between the degree of collagen VI deficiency and the clinical severity in UCMD. No significant phenotypical differences were found between the families linked to each of the 2 loci, which confirms UCMD as a unique entity with underlying genetic heterogeneity. PMID- 15127310 TI - The spectrum of cranial ultrasound and magnetic resonance imaging abnormalities in congenital cytomegalovirus infection. AB - Congenital cytomegalovirus (CMV) infection can lead to severe neurological sequelae and (progressive) sensorineural deafness. Neonatal imaging data is mainly based on cranial ultrasound (US) and computed tomography (CT). The additional value of magnetic resonance imaging (MRI) was assessed in congenital CMV infection. The eleven infants studied had a gestational age between 34 and 41 weeks and a birth weight between 1000 and 2780 grams. All but 2 of the infants presented with microcephaly and jaundice at birth. The diagnosis was confirmed postnatally in all infants by isolation of the virus or a polymerase chain reaction (PCR) from the urine. Cranial US was performed in all, MRI in 6 during the neonatal period and later in infancy in 2. Auditory brainstem evoked responses (ABR) were performed in all survivors. US showed periventricular calcifications and/or lenticulostriate vasculopathy associated with mild to moderate ventricular dilatation in 10 of the 11 children. Periventricular (pseudo) cysts were seen in 6 children, being occipital in 4, temporal in 3 and fronto-parietal in 1. The cerebellum appeared to be small in 4 children. MRI provided additional information in 6 of the 8 children. Polymicrogyria in the perisylvian region was seen in 4 children, hippocampal dysplasia in 3 and cerebellar hypoplasia in 4 children. Abnormal signal intensity in the white matter was seen in 4 infants. ABRs were abnormal in 7 of the 9 children. Four children died in the neonatal period, 4 developed severe neurological sequelae, associated with epilepsy and sensorineural deafness in 3. Three children were still too young to be tested, but 2 of these showed sensorineural deafness. MRI provided important additional information, especially with regard to associated polymicrogyria, hippocampal dysplasia, and cerebellar hypoplasia. Calcifications were better seen using US. A combination of US and neonatal MRI should be recommended instead of a CT which is still recommended in the literature. PMID- 15127311 TI - Low level of intracortical inhibition in children shown by transcranial magnetic stimulation. AB - Transcranial magnetic stimulation (TMS) is an established neurophysiological tool to evaluate the integrity and maturation of the corticospinal tract. TMS was used in this study to compare intracortical inhibition (ICI) in children, adolescents, and adults. The paired-pulse technique of TMS with interstimulus intervals of 2 ms was used to determine the ratio of conditioned (cMEP) and unconditioned amplitudes (ucMEP) that measures ICI. In experiment 1 (Exp 1) stimulus intensity was adapted to motor threshold (50 healthy subjects; 24 male, 26 female, median age 13.5 years, range 6.3 - 34 years) and in experiment 2 (Exp 2) stimulus intensity was adapted to the ucMEP (200 - 400 microV). Children (quotient of cMEP and ucMEP: Exp. 1: 0.71 +/- 0.41, Exp. 2: 0.82 +/- 0.25) had significantly less ICI compared to adults (Exp. 1: 0.21 +/- 0.19, mean +/- STD, Exp. 2: 0.35 +/- 0.22, in both experiments p < 0.001). Recently, ICI has been linked to the regulating function of GABAergic cortical interneurons on practice-dependent neuronal plasticity. Therefore, the lower ICI in children points to maturation processes that may have implications for the greater capacity of practice dependent neuronal plasticity in children. PMID- 15127312 TI - Effect of doxapram on cerebral blood flow velocity in preterm infants. AB - BACKGROUND: Doxapram is used to treat apnea of prematurity when there is an insufficient response to methylxanthine treatment. As an unwanted side effect, reduced cerebral perfusion has been seen in methylxanthine-treated infants while effects of doxapram on the cerebral perfusion have not been studied yet. PATIENTS AND METHODS: Fifteen preterm infants treated with doxapram were included in the study. Birth weight ranged from 380 g to 1150 g (median 740 g), gestational age from 24 to 27 weeks (median 26 weeks). Infants received a doxapram loading dose (2.5 mg/kg) over a 30-minute period, followed by a continuous infusion of 0.5 mg/kg/h. Using Doppler sonography, blood flow velocities and the resistance index were measured in the anterior cerebral artery. Measurements were performed at baseline and 30 and 120 minutes after the start of doxapram. RESULTS: Maximal systolic blood flow velocity (V(max)) decreased significantly after the infants had received the loading dose (V(max) baseline: 40.7 cm/s +/- 6.9 [mean +/- SD]; V(max) 30 min: 35 cm/s +/- 8.9; p = 0.0017) but returned to near baseline values at 120 min (38.5 +/- 9.0, p = 0.22). End-diastolic, time-averaged, and time averaged maximal velocities did not change significantly at 30 or 120 min. CONCLUSIONS: Doxapram induced a significant decrease in maximal cerebral blood flow velocity. Further studies are needed to assess whether this decrease may be critical to cerebral white matter perfusion in the vulnerable preterm infant. PMID- 15127313 TI - Magnetic resonance imaging for the diagnosis of acute leukoencephalopathy in children treated with tacrolimus. AB - The leukoencephalopathy induced by tacrolimus is increasingly recognised as an important cause of neurological complications after transplantation. Magnetic resonance imaging (MRI) is of major help in the differential diagnosis of infection or vascular injury. We describe two children with coma and seizures after transplantation, in whom the results of MRI with FLAIR (fluid-attenuated inversion-recovery) and DWI (diffusion-weighted images) were the main positive elements for the diagnosis of drug-induced toxicity. The results of DWI favoured the role of oedema and/or demyelination in the pathophysiology of this disorder. Unlike other reported patients, in whom all symptoms resolved, lesions persisted, albeit improved, on the control MRI, and both children demonstrated learning disabilities after several years of follow-up. PMID- 15127314 TI - Acquired torticollis due to Grisel's syndrome: case report and follow-up of non traumatic atlantoaxial rotatory subluxation. AB - Non-traumatic atlantoaxial rotatory subluxation is an uncommon entity, with inconsistent presentations. It is also known as Grisel's syndrome (GS), and most commonly follows infectious processes of the upper respiratory tract. A case is described of a non-traumatic rotatory atlantoaxial dislocation in a three-year old boy. The patient presented with acute torticollis one week after mild upper respiratory infection. Neurological evaluation and lumbar punction were normal. After five days of intractable pain and non-reducible rotational tilt of the head to the left, CT and MR imaging were performed and showed atlantoaxial rotatory subluxation. In addition, MRI demonstrated middle ear and mastoid inflammation. Tilt reduction could be obtained through gentle skull traction under sedation and relaxation for 3 days. Antibiotic treatment was performed. The patient was then placed in a Minerva cast jacket. After seven weeks of immobilization, CT demonstrated regular atlantoaxial alignment. Follow-up after 5.5 years showed the boy neurologically intact and free of clinical complaints. Dynamic cervical radiographs confirmed that the atlantoaxial joints were stable. PMID- 15127315 TI - Neonatal seizures in two sisters with incontinentia pigmenti. AB - Familial incontinentia pigmenti (IP) (OMIM #308300) is a rare genetic disorder which segregates in an X-linked dominant way. The female-to-male ratio ranges from 20 to 37 : 1. In affected females IP causes highly variable abnormalities of the skin, hair, nails, teeth, eyes, and central nervous system (CNS). Cardiovascular anomalies, cerebral infarction, and immune dysfunction are rare complications of IP. The pathogenesis of cerebral changes in IP remains elusive. We report the case of two IP-affected sisters who presented in each case with neonatal seizures on the fifth day of life. Via cranial magnetic resonance tomographic imaging (MRI) different types of lesions in both hemispheres were demonstrable in both patients. To date the pathogenetic mechanisms for the cerebral lesions are not fully understood. However, multiple microscopic infarcts could serve as a possible explanation. The clinical course and the neurological development of the older child are favorable and so far the younger sibling appears to be developing normally, which is uncommon for patients with early onset of neurological symptoms. Symptomatic seizures in IP are an important differential diagnosis in benign non-familial and familial neonatal seizures. PMID- 15127316 TI - Acute haemorrhagic encephalomyelitis (AHEM): MRI findings. AB - Acute disseminated encephalomyelitis (ADEM) comprises a group of demyelinating disorders of the CNS, of which a haemorrhagic variant indicates the most severe clinical course (AHEM). It is supposed to be caused by an overshooting immunological response following an infection or a vaccination. Restricted diffusion of protons could be demonstrated using DW-MRI in a case of rapidly progressive disease with additional haemorrhage. Hence, the addition of DWI adds to the diagnostic power of MRI in the setting of post-infectious demyelinating disorders of the CNS. PMID- 15127317 TI - Episodic ataxia type 1 with distal weakness: a novel manifestation of a potassium channelopathy. AB - Episodic ataxia type1 (EA1) is an autosomal dominant disorder characterised by episodes of ataxia, dysarthria, tremor and visual disturbances lasting for seconds or minutes, precipitated by physical and emotional stress, startle or sudden movements. In addition there is continuous myokymia. Phenotypic variants such as the combination with epilepsy, shortening of the Achilles tendon in children, transient postural abnormalities in infancy, and a very few patients with longer lasting episodes have been reported. We describe a 10-year-old girl with EA1 who has distal weakness with paresis of the extensors of the feet and prolonged spells of limb stiffness (neuromyotonia) lasting up to 12 hours. A novel single nucleotide change at position 785 T > C that alters a highly conserved residue in the third transmembrane segment of the voltage-gated potassium channel Kv1.1 was found. PMID- 15127318 TI - Diabetes and cardiovascular risk evaluation and management in primary care: progress and unresolved issues - rationale for a nationwide primary care project in Germany. AB - This review highlights established and more recently recognized risk factors for coronary heart disease (CHD) relevant for patients seen in primary care, emphasizing the key role of diabetes mellitus type 2. Recent trends in risk factor research as well as current methods of risk stratification, and new systemic markers are discussed. Beyond the need for more forceful public health strategies to improve early recognition and intervention, the necessity of an integrated comprehensive investigation of the overall characteristics of cardiovascular disease, especially in primary care patients as a prerequisite for future concerted actions is pointed out. Based on this, a large-scale epidemiological investigation focusing on CHD and diabetes in the primary care sector is suggested. PMID- 15127319 TI - TSH-receptor autoantibodies - differentiation of hyperthyroidism between Graves' disease and toxic multinodular goitre. AB - Previous studies indicate pre-existing subclinical Graves' disease in many patients with the scintigraphic diagnosis of toxic multinodular goitre type A, equivalent to the in Germany so-called disseminated thyroid autonomy. Furthermore, after radioiodine treatment an increase or the induction of TSH receptor antibodies (TRAb) in patients with Graves' disease or toxic multinodular goitre has been repeatedly reported. The distinction between both hyperthyroid conditions, Graves' disease and toxic multinodular goitre type A, depends on the diagnostic power of the TSH-receptor antibody determination. Bioassays using CHO cell lines expressing the hTSH-receptor or a new TBII assay based on competitive binding to recombinant human TSH-receptor showed a higher sensitivity for the detection of TSH-receptor antibodies in patients with Graves' disease than previous assays using solubilized porcine epithelial cell membranes. In up to 50 % of patients with toxic multinodular goitre A without antithyroid drug pretreatment TSH-receptor antibodies were detectable with a high correlation between thyroid-stimulating antibodies in the bioassay and the h-TBII assay. Moreover, in a recent study the development of TSH-receptor antibodies after radioiodine treatment was detectable in 36 % of patients with toxic multinodular goitre type A, whereas TSH-receptor antibodies were not detectable in patients with toxic multinodular goitre type B or in patients with toxic adenoma. In conclusion, thyroid-stimulating antibodies in a bioassay or TSH-receptor antibodies detected with the h-TBII assay have the highest diagnostic power to differentiate Graves' disease from toxic multinodular goitre. Because of its less cumbersome assay technique the h-TBII should be performed in all patients with hyperthyroidism to differentiate Graves' disease from non-autoimmune hyperthyroidism such as toxic multinodular goitre to select the appropriate therapy for these patients. PMID- 15127320 TI - Enhancement of fibrinolysis in poorly controlled, hospitalized type 2 diabetic patients by short-term metabolic control: association with a decrease in plasminogen activator inhibitor 1. AB - Impaired fibrinolysis in type 2 diabetes may be caused by an increased plasma concentration of plasminogen activator inhibitor 1 (PAI-1), although the effects of short-term hypoglycemic therapy on fibrinolytic activity are poorly understood. This study investigated the effects of metabolic improvement on fibrinolysis activity and plasma concentrations of PAI-1 in poorly controlled, hospitalized type 2 diabetic patients. Forty-eight poorly controlled type 2 diabetic patients were studied; 26 were subsequently treated with sulfonylurea (SU) and 22 with insulin. The plasma concentrations of plasmin-alpha2-antiplasmin (PAP), a measure of fibrinolytic activity, plasma PAI-1, and fasting triglycerides and glucoses were measured at the beginning and the end of hospitalization. The body mass index and fasting triglyceride decreased significantly after treatment (p < 0.0001). The plasma concentration of PAP increased significantly (p < 0.01), and the plasma PAI-1 decreased by 50% after treatment. There was an inverse correlation between the changes in the plasma concentrations of PAP and PAI-1 (r= - 0.36, p = 0.023). Treatment with SU or insulin showed an increase in plasma PAP with a concomitant decrease in the plasma PAI-1 with equivalent glycemic control. In poorly controlled type 2 diabetic patients, the plasma PAP concentration can be significantly increased and the plasma PAI-1 antigen significantly reduced, even with short-term metabolic improvements including weight reduction, a better lipid profile, and tighter glycemic control with either SU or insulin therapy, and that enhanced fibrinolysis may be mediated partly through a decrease in the plasma PAI-1 after metabolic control. PMID- 15127321 TI - Beta-cell secretory function and CD25 + lymphocyte subsets in the early stage of type 1 diabetes mellitus. AB - Cellular immunologic tests have not been used for diagnostic purposes in individuals at risk for autoimmune insulitis or in patients with partial beta cell destruction because of a lack of studies that show their predictive value. In this study we initially evaluated 43 patients with recent-onset Type 1 diabetes (disease duration 100 150 200 mm (2)). Ulcer healing was assessed by planimetric measurement of the wound area every second week until wound healing. The time course of wound healing in the different groups was compared by the weekly wound radius reduction using the equation R = sqrt A/pi. The average healing time in group A was 70 (95 %-CI 64 - 77) days with a wound radius reduction of 0.42 mm/week (95 %-CI 0.28 - 0.56). In group B the average healing time was 79 (95 %-CI 75 - 82) days and the weekly wound radius reduction was 0.47 mm (95 %-CI 0.45 - 0.49). The average healing time in group C was 85 (95 %-CI 80 - 89) days with a wound radius reduction of 0.53 mm/week (95 %-CI 0.42 - 0.56). In group D the average healing time was 97 (95 %-CI 91 - 103) days. The weekly wound radius reduction was 0.57 mm (95 %-CI 0.49 - 0.81). Wound radius reductions and the time needed for healing are affected by the ulcer area, a measure of ulcer size, in neuropathic diabetic foot ulcers. The calculation of the weekly wound radius reduction for different ulcer areas may be a useful tool in daily clinical practice to identify ulcers who do not respond adequately to the treatment. PMID- 15127324 TI - Beta2-microglobulin and cystatin C in type 2 diabetes: assessment of diabetic nephropathy. AB - BACKGROUND: Changes in glomerular filtration rate (GFR) provide a valuable indicator of the progression of diabetic nephropathy (DN). This study was designed to demonstrate the clinical values of serum cystatin C (Cys C) and beta2 microglobulin in the assessment of renal function in type 2 diabetics by comparing them with the GFR, estimated from the uptake phase of 99 m technetium dimetiltriamino pentaacetic acid renogram (GFR-DTPA) and creatinine clearances. MATERIALS AND METHODS: 68 type 2 diabetic patients with (urinary albumin excretions (UAE) 30 - 300 mg/24 h) (n = 39) and without (UAE < 30 mg/24 h) (n = 29) microalbuminuria and 32 controls were enrolled in the study. Serum Cys C, beta2-microglobulin, creatinine, urinary microalbumin levels, creatinine clearances and GFR-DTPA values were determined in all groups. Non-parametric ROC curves, using a cut-off GFR-DTPA of 60 mL/min/1.73 m (2), were obtained for these markers. RESULTS: Serum Cys C, beta2-microglobulin, glucose and HbA1c concentrations were significantly higher in the group with diabetes compared to controls. In the patients with microalbuminuria, serum Cys C and glucose concentrations increased significantly in comparison to patients with normoalbuminuria, while no differences were observed for beta2-microglobulin levels. Serum creatinine concentrations, GFR-DTPA values and creatinine clearances were not different between both diabetic groups and controls. Cys C was positively correlated with beta2-microglobulin and creatinine and negatively with GFR values; beta2-microglobulin was also positively correlated with serum creatinine in microalbuminurics. A significant inverse correlation was found between beta2-microglobulin and GFR values in both microalbuminurics and normoalbuminurics. CONCLUSIONS: Increased Cys C and beta2-microglobulin in diabetics may be early indicators of incipient DN. The diagnostic accuracies of Cys C and beta2-microglobulin are superior to that of serum creatinine in distinguishing between mild and moderately reduced GFR. PMID- 15127325 TI - Weight reduction by sibutramine in obese subjects in primary care medicine: the SAT Study. AB - BACKGROUND: Sibutramine is a selective serotonin and noradrenaline reuptake inhibitor that is known to reduce body weight. The efficacy of this drug in primary care medicine is currently unknown. OBJECTIVE: To study, in a primary healthcare setting, the effect of a standardized non-pharmacological treatment program and 15 mg sibutramine or placebo on long-term weight reduction in obese subjects with a body mass index >or= 30 and < 40 kg/m(2). DESIGN: A multicentre, double-blind, placebo-controlled, randomized, parallel group comparison over 54 weeks of continuous therapy. SETTING: 33 general practitioners in Germany. SUBJECTS: 389 obese patients were recruited of whom 362 were randomized. MEASUREMENTS: Primary measure was weight reduction at week 54; others included reduction in BMI, waist circumference, waist-hip ratio, blood pressure and blood lipids. RESULTS: 348 obese subjects were analyzed using an intention-to-treat analysis. Mean weight loss in the sibutramine (S) group was 8.1 +/- 8.2 kg vs. 5.1 +/- 6.5 kg in the placebo (P) group (p < 0.001; Intent-to-treat analysis). More subjects lost more than 5 % and 10 % of initial weight with sibutramine than with placebo (5 %, S: 62.6 %, P: 41.4 %, p < 0.001; 10 %, S: 40.8, P: 19.0 %, p < 0.001). Weight loss was accompanied by an improvement in the lipid profile, in particular, an increase in HDL-cholesterol and a decrease in fasting triglycerides. In both groups, systolic and diastolic blood pressure decreased in those with moderate hypertension and remained unchanged in those with normal blood pressure at baseline. There was a modest increase in heart rate in S (1.9 beats/min) vs. P (- 0.9 beats/min) (p < 0.05). CONCLUSIONS: Under primary care conditions, sibutramine 15 mg daily proved to be a safe and effective drug for additional weight loss in obese subjects undergoing a comprehensive weight reduction program. PMID- 15127326 TI - Differential regulation of ANG2 and VEGF-A in human granulosa lutein cells by choriogonadotropin. AB - The growth and development of the corpus luteum after rupture of the follicle is a highly regulated process characterised by a rapid vascularization of the follicle surrounding granulosa cells. Vascularization is regulated by a large number of growth factors and cytokines whereas members of the angiopoietin family and VEGF-A are reported to play a principal role. The gonadotropic hormones luteinizing hormone and choriogonadotropin are reported to be essential for corpus luteum formation. In this study we investigated by RT PCR if the growth factors PGF, PDGF-A, PDGF-B, VEGF-A, VEGF-B, VEGF-C, VEGF-D, ANG1, ANG2, ANG3 and ANG4 are expressed in granulosa cells. We show the expression of VEGF-A, VEGF-B, PDGF-A, ANG1 and ANG2 in granulosa cells. Using RT-PCR and Real-Time PCR we demonstrate that angiopoietin 2 is downregulated in human granulosa cells in vitro after choriogonadotropin treatment whereas the expression of angiopoietin 1 is not significantly altered. The expression of VEGF on the RNA- and on the protein level was determined. It was shown that in granulosa cells VEGF is upregulated after choriogonadotropin treatment on the RNA level and that increasing concentrations of choriogonadotropin from 0 to 10 U/ml leads to an increasing amount of VEGF in the cell culture supernatants. The amount of VEGF in the supernatants reaches a plateau at 0.5 U/ml and is increased only slightly and not significantly after treatment of the cells with 10 U/ml choriogonadotropin compared to 0.5 U/ml. In total these findings suggests that in granulosa cells the mRNA of various growth factors is detectable by RT-PCR and that VEGF-A and ANG2 is regulated by the gonadotropic hormone choriogonadotropin. These findings may add impact on the hypothesis of choriogonadotropin as a novel angiogenic factor. PMID- 15127327 TI - The effects of prolonged, intracerebroventricular prolactin treatment on luteinizing hormone secretion, catecholaminergic activity and estrous behavior in ewes. AB - Searching for the role of prolactin (PRL) in controlling gonadotropic axis activity in sheep, we studied the effects of prolonged, intracerebroventricular (i.c.v.) PRL infusion on luteinizing hormone (LH) secretion and catecholaminergic activity in the hypothalamic infundibular nuclei/median eminence (IN/ME) in sexually active ewes during the periovulatory period. Three groups of animals received the following treatments: 1). i.c.v. infusion of PRL at a dose of 200 microg/day (Lower dose, n = 5); 2). i.c.v. infusion of PRL at a dose of 400 microg/day (Higher dose, n = 6), and 3). i.c.v. infusion of the vehicle (control, n = 5). Each dose of PRL was infused in a pulsatile manner, 4 x 50 microg/h and 4 x 100 microg/h, in 30-min intervals, respectively, during four consecutive days before oncoming ovulation. The estrous behavior of ewes following treatments was also monitored as a determinant of the GnRH/LH surge. Two series of blood collections were made in every ewe, the first on the day preceding the infusion (day 0 of the experiment), the second on the day after the infusion (day 5 of the experiment). In addition, on day 5 of the experiment, perfusions of the IN/ME were made by the push-pull method, either in control or lower dose-treated animals. It was shown that a significant (p < 0.01, p < 0.001) increase in tonic LH secretion during the periovulatory period remained in ewes irrespective of the kind of infusion. No statistical differences were found in LH pulse frequency, amplitude, or in the length of the pulse when compared with values from day 0 and 5 of the experiment within each group. A significant (p < 0.001) increase in IN/ME perfusate concentrations of dopamine and noradrenaline metabolites was noted in PRL-treated ewes in comparison with those in the control. The estrous behavior in PRL-treated animals was delayed for a few days, 3.80 +/- 0.80 days at the lower dose (p < 0.01), and 2.83 +/- 0.98 days at the higher dose (p < 0.05) in comparison with the control, 0.20 +/- 0.20 days. These data indicate that maintenance of an increased PRL concentration within the central nervous system (CNS) for a few days before oncoming ovulation has no inhibitory effect on tonic LH secretion. A few-day shift of the preovulatory GnRH/LH surge, as determined by estrous behavior, might, however, be a consequence of the PRL-induced increase in catecholamine turnover in the IN/ME. PMID- 15127328 TI - Characterization of group B streptococci recovered from infants with invasive disease in England and Wales. AB - Group B streptococci (GBS) are a major cause of invasive disease in infants, with enhanced surveillance in England and Wales showing an incidence of 0.74 cases per 1000 live births and a mortality rate of 8%. Among 353 isolates obtained during enhanced surveillance, the predominant serotypes were III (48%), Ia (27%), and V (10%), and the remainder comprised Ib, II, IV, VI, and VII; 3% were not typable. Isolates from patients with early-onset disease had serotypes III (38%), Ia (32%), and V (13%), with late-onset disease having a higher incidence of type III (67%) strains. Patients infected with serotype III strains had a higher rate of meningitis, and those with type V strains had a higher mortality rate. Isolates were susceptible to penicillin and ampicillin, but 4% were resistant to erythromycin, and 91% were resistant to tetracycline. A trivalent vaccine containing capsular polysaccharides III, Ia, and V could theoretically provide coverage against 85% of the cases of GBS disease among infants in England and Wales. PMID- 15127329 TI - Easing the burden: characterizing the disease burden of neonatal group B streptococcal disease to motivate prevention. PMID- 15127330 TI - Why don't they listen? Adherence to recommendations of infectious disease consultations. AB - The effectiveness of an infectious diseases (ID) consultation is dependent on adherence to the recommendations. To delineate the factors that affect adherence, we conducted a prospective cohort study of 465 consultations at 2 academic institutions in which we evaluated the consultation process, patient and consultant characteristics, types of recommendations, and compliance with recommendations. The overall compliance rate was 80%, with 85% adherence to crucial recommendations. Multivariate analysis revealed that adherence to ID recommendations was higher when the recommendations were therapeutic instead of diagnostic, when they related to a specific clinical question, when recommendations were deemed crucial by the ID service, if the primary service was medicine, and if the consultation note was legible and organized. Whether modification of consultant practice will lead to improved recommendation compliance and patient outcomes warrants further study. PMID- 15127331 TI - Infectious diseases consultative recommendations: if heard, they can be listened to. PMID- 15127332 TI - Two subtypes of genotype B (Ba and Bj) of hepatitis B virus in Japan. AB - We have previously reported 2 subtypes of hepatitis B virus (HBV) genotype B, one of which has the recombination with genotype C over the precore region plus core gene (Ba) and the other of which does not (Bj). A restriction fragment-length polymorphism method with 2 endonucleases was newly developed for distinguishing between subtypes Ba and Bj and was applied to 313 carriers of HBV genotype B in Japan. Subtype Ba was detected in 38 (12%) and subtype Bj in 275 (88%) of the carriers of HBV genotype B. Hepatitis B e antigen in serum was found more frequently in patients with chronic infection with subtype Ba than in those with chronic infection with subtype Bj (8 [32%] of 25 vs. 25 [9%] of 273; P<.01). The new method for distinguishing between Ba and Bj by restriction fragment-length polymorphism would be useful in examining the distribution of these 2 subtypes in situations in which HBV genotype B is prevalent. PMID- 15127333 TI - Mycobacterial infection: a difficult and late diagnosis in stem cell transplant recipients. AB - The Infectious Diseases Working Party of the European Blood and Marrow Transplant Group conducted a survey to obtain information about the frequency, presentation, and treatment of mycobacterial infection (MBI) in stem cell transplant (SCT) recipients. Among 29 centers, MBI was diagnosed in 0.79% of 1513 allogeneic and 0.23% of 3012 autologous SCT recipients during 1994-1998 a median of 160 days after transplantation. The mean interval between first symptoms and diagnosis was 29 days and was still longer for patients with atypical MBI or recipients of corticosteroid therapy. The prevalence of MBI was highest among those who received matched unrelated or mismatched STCs from related donors. Of 31 patients, 20 had tuberculosis, 8 had atypical MBI, and 3 had diagnoses based on histological findings only. Five patients (16%) died, all of whom had received an allogeneic SCT. Because of the increased numbers of unmatched donors and transplantation programs in countries with a high prevalence of tuberculosis, constant vigilance is required to early detect MBI in SCT recipients. PMID- 15127334 TI - Fusarium infection in hematopoietic stem cell transplant recipients. AB - To characterize the epidemiology and prognostic factors of invasive fusariosis in hematopoietic stem cell transplant (HSCT) recipients, the records of HSCT recipients from 9 hospitals (7 in Brazil and 2 in the United States) were retrospectively reviewed. Sixty-one cases were identified: 54 in allogeneic HSCT recipients and 7 in autologous HSCT recipients. The incidence of fusariosis among allogeneic HSCT recipients varied between a range of 4.21-5.0 cases per 1000 in human leukocyte antigen (HLA)--matched related transplant recipients to 20.19 cases per 1000 in HLA-mismatched transplant recipients. The median time period between transplantation and diagnosis of fusariosis was 48 days. Among allogeneic HSCT recipients, a trimodal distribution was observed: a first peak before engraftment, a second peak at a median of 62 days after transplantation, and a third peak >1 year after transplantation. The actuarial survival was 13% (median, 13 days). Persistent neutropenia was the single prognostic factor for death identified by multivariate analysis. PMID- 15127335 TI - Virulence associated with outbreak-related strains of Burkholderia cepacia complex among a cohort of patients with bacteremia. AB - The Burkholderia cepacia complex includes 9 genomovars. The relative virulence of each is unknown. Host and pathogen features associated with mortality were evaluated among patients with B. cepacia complex bacteremia. Cases were ascertained through review of blood culture results for the period of May 1996 through May 2002. Isolates were identified to species level with 16S rDNA and recA-based species-specific polymerase chain reaction analyses and recA restriction fragment-length polymorphism. Strain typing was performed with pulsed field gel electrophoresis. Fifty-three patients with B. cepacia complex bacteremia were identified; only 9 (17%) had cystic fibrosis. Twenty-five patients (47%) died within 14 days of bacteremia. After controlling for comorbid conditions and therapeutic interventions, 2 outbreak-related strains of Burkholderia cenocepacia (genomovar III) were associated with 14-day mortality (odds ratio, 5.5; 95% confidence interval, 1.20-25.02). B. cenocepacia is an emerging nosocomial pathogen. Certain strains are associated with an enhanced capacity for interpatient spread and poor outcome. PMID- 15127336 TI - Effect of macrolide consumption on erythromycin resistance in Streptococcus pyogenes in Finland in 1997-2001. AB - The aim of this study was to investigate the association between regional macrolide resistance in Streptococcus pyogenes and macrolide use in Finland. During 1997-2001, a total of 50,875 S. pyogenes isolates were tested for erythromycin susceptibility in clinical microbiology laboratories throughout Finland. The local erythromycin resistance levels were compared with the regional consumption data of all macrolides pooled and, separately, with the use of azithromycin. The regional resistance rates of 1 year were compared with the regional consumption of the previous year and with the average rates of use for the 2 previous years. A linear mixed model for repeated measures was used in modeling the association. A statistically significant association existed between regional erythromycin resistance in S. pyogenes and consumption of macrolides; association with azithromycin use alone was not found. PMID- 15127337 TI - West Nile virus-associated encephalitis in recipients of renal and pancreas transplants: case series and literature review. AB - Although West Nile fever is mild in the vast majority of infected persons, there is growing evidence that the disease may be more severe in the immunocompromised population. We describe 3 recipients of kidney or pancreas transplants who developed West Nile fever, 2 of whom had meningoencephalitis. As is the norm when treating serious infections in transplant recipients, a reduction of immunosuppression was pursued for these patients. Despite the severe nature of the disease in 2 patients, all recovered from the disease. The time course of neurologic recovery in the 2 patients with meningoencephalitis is highlighted. We also review the literature on West Nile fever in organ transplant recipients. In areas where West Nile virus is endemic, one must have a high index of suspicion for the illness when dealing with fever in transplant recipients. PMID- 15127338 TI - Granulomatous infectious diseases associated with tumor necrosis factor antagonists. AB - The relationship between the use of tumor necrosis factor antagonists and onset of granulomatous infection was examined using data collected through the Adverse Event Reporting System of the US Food and Drug Administration for January 1998 September 2002. Granulomatous infections were reported at rates of approximately 239 per 100,000 patients who received infliximab and approximately 74 per 100,000 patients who received etanercept (P<.001). Tuberculosis was the most frequently reported disease, occurring in approximately 144 and approximately 35 per 100,000 infliximab-treated and etanercept-treated patients, respectively (P<.001). Candidiasis, coccidioidomycosis, histoplasmosis, listeriosis, nocardiosis, and infections due to nontuberculous mycobacteria were reported with significantly greater frequency among infliximab-treated patients. Seventy-two percent of these infection occurred < or =90 days after starting infliximab treatment, and 28% occurred after starting etanercept treatment (P<.001). These data indicate a risk of granulomatous infection that was 3.25-fold greater among patients who received infliximab than among those who received etanercept. The clustering of reports shortly after initiation of treatment with infliximab is consistent with reactivation of latent infection. PMID- 15127339 TI - Miltefosine for new world cutaneous leishmaniasis. AB - The oral agent miltefosine has demonstrated a >95% cure rate in Indian visceral leishmaniasis. We performed a large, placebo-controlled study of miltefosine therapy (2.5 mg/kg per day orally for 28 days) against cutaneous leishmaniasis in Colombia and Guatemala. In regions in Colombia where Leishmania vianna panamensis is common, the per-protocol cure rates for miltefosine and placebo were 91% (40 of 44 patients) and 38% (9 of 24). These values are similar to historic values for the antimony standard of care and placebo. In regions in Guatemala where L. v. braziliensis and L. mexicana mexicana are common, the per-protocol cure rates were 53% (20 of 38) for miltefosine and 21% (4 of 19) for placebo. The miltefosine rate was lower than historic antimony cure rates of >90%. Miltefosine was well tolerated. Miltefosine is a useful oral agent against cutaneous leishmaniasis due to L. v. panamensis in Colombia but not against leishmaniasis due to L. v. braziliensis in Guatemala. PMID- 15127340 TI - Pneumococcal endocarditis in children. AB - Endocarditis due to Streptococcus pneumoniae is unusual in children, accounting for 3%-7% of all cases of childhood endocarditis. The US Pediatric Multicenter Pneumococcal Surveillance Group has prospectively identified patients with invasive disease at 8 children's hospitals. During the period of 1 September 1993 through 28 February 2003, a total of 11 children with pneumococcal endocarditis were seen. Seven (64%) were 3-36 months old; 8 (73%) were boys. Ten (91%) had preexisting structural heart disease; 5 had undergone previous heart surgery. Concomitant sites of infection were noted in 6 patients (55%), including 3 patients with meningitis. One patient (9%) died during hospitalization, and 5 others (45%) experienced serious complications. Only 2 patients remained hospitalized for their entire course of parenteral antibiotic therapy. Eight of 10 pneumococcal isolates tested were vaccine or vaccine-related serotypes included in the currently licensed 7-valent conjugated pneumococcal vaccine. Pneumococcal endocarditis in children is unusual but often has serious complications. PMID- 15127341 TI - Trends in antimicrobial drug development: implications for the future. AB - The need for new antimicrobial agents is greater than ever because of the emergence of multidrug resistance in common pathogens, the rapid emergence of new infections, and the potential for use of multidrug-resistant agents in bioweapons. Paradoxically, some pharmaceutical companies have indicated that they are curtailing anti-infective research programs. We evaluated the United States Food and Drug Administration (FDA) databases of approved drugs and the research and development programs of the world's largest pharmaceutical and biotechnology companies to document trends in the development of new antimicrobial agents. FDA approval of new antibacterial agents decreased by 56% over the past 20 years (1998-2002 vs. 1983-1987). Projecting future development, new antibacterial agents constitute 6 of 506 drugs disclosed in the developmental programs of the largest pharmaceutical and biotechnology companies. Despite the critical need for new antimicrobial agents, the development of these agents is declining. Solutions encouraging and facilitating the development of new antimicrobial agents are needed. PMID- 15127342 TI - Are antimicrobial-impregnated catheters effective? Don't throw out the baby with the bathwater. AB - The antimicrobial-impregnated central venous catheter (CVC) has been the most intensively studied technology for the prevention of CVC-related bloodstream infections (BSIs) over the past 30 years. Although more than a dozen randomized trials have shown significant benefit, authors of an analysis published in a recent issue of Clinical Infectious Diseases have raised questions about the efficacy of antimicrobial-impregnated CVCs because of perceived defects in the experimental design of the studies and statistical analyses of the data. They have further argued that even if this technology might be effective in preventing CVC-related BSI, its cost-effectiveness is questionable. Although most of the studies scrutinized by the authors of this analysis indeed had shortcomings, we believe that their analysis unjustifiably downplays a large body of research that has demonstrated a consistent reduction in CVC-related BSI and a clear-cut cost effectiveness associated with the use of antimicrobial-impregnated CVCs. PMID- 15127343 TI - Use of a broader determinants of health model for community-acquired pneumonia in seniors. AB - Community-acquired pneumonia in older adults represents an important clinical and public health challenge. This article discusses the role that factors such as socioeconomic status, air pollution, crowding, exposure to tobacco smoke, and nutrition play in predisposing elderly persons to such respiratory infections. It is proposed that a model that addresses these factors is needed for a comprehensive understanding of these infections. Although the causal pathways may be unclear, there are data to suggest a relationship between low socioeconomic status and risk of acquiring respiratory infection. The need for more research in this area is emphasized. PMID- 15127344 TI - Shiga toxin-producing Escherichia coli infection. AB - Large-scale outbreaks of Shiga toxin-producing Escherichia coli (STEC) infection have revealed the great disease-causing potential of this organism, especially among children and elderly persons. Approximately 5%-10% of people with STEC infection will develop hemolytic-uremic syndrome (HUS), approximately 10% of those who develop HUS will die or have permanent renal failure, and up to 50% of those who develop HUS will develop some degree of renal impairment. Important concepts in understanding the pathogenesis and prevention of STEC-associated HUS are emerging, although no specific therapy yet exists. Optimal management of STEC infection includes intravenous hydration, avoidance of antimotility agents and antimicrobials, and monitoring for sequelae. Antimicrobials may have a potentially harmful role, possibly by inducing intestinal production of Shiga toxin during the diarrheal phase of illness. A recent clinical trial evaluating an intraluminal Shiga toxin-binding agent to ameliorate HUS showed no improvement in outcome. Interventions to prevent HUS from developing in STEC-infected children are under investigation. Prevention of exposure to STEC remains important, and animal vaccines to prevent stool shedding of STEC among food animals are in development. PMID- 15127345 TI - Guide to sexually transmitted disease resources on the Internet. AB - The Internet provides patients, clinicians, teachers, and researchers with immediate access to reliable information, authoritative recommendations, and the latest research findings and statistics, but quickly finding the best sources while avoiding the unreliable and obsolete can be a problem. We searched the Internet for the most useful English-language Web sites on sexually transmitted diseases (STDs), with annotations, in 4 tables: sites for patients, for clinicians and teachers, and for researchers, and sites dedicated to a single STD. In the process, we found that government-sponsored sites tended to have the most reliable information. This held true regardless of the kind of information we were seeking. Several university-sponsored sites contained information that was outdated or erroneous. Commercial and nonprofit sites sometimes evinced a bias that could mislead some readers. Both health care professionals and laypersons seeking information about STDs on the World Wide Web should generally start their search at government-sponsored sites. PMID- 15127346 TI - Predictors of virologic failure and resistance in HIV-infected patients treated with nevirapine- or efavirenz-based antiretroviral therapy. AB - Resistance to nonnucleoside reverse transcriptase inhibitors (NNRTIs) increases with the wider use of this class of antiretroviral therapy. The association between adherence and resistance to NNRTI-based regimens is poorly understood. Predictors of virologic failure and resistance according to a baseline evaluation of nonadherence risk factors were determined in a cohort of 71 human immunodeficiency virus (HIV)-infected patients with early virologic response who received an NNRTI-based regimen. During the median follow-up of 29 months, 20 (28%) of 71 patients experienced virologic failure with an NNRTI-based regimen. Virologic failure was associated with repeated drug holidays (> or =48 h of unplanned drug cessation), depression, younger age, and low adherence to therapy during baseline evaluation. Moreover, repeated drug holidays was the only risk factor for developing a major mutation conferring cross-resistance to the NNRTI class (hazard ratio, 22.5; 95% confidence interval, 2.8-180.3; P<.0001). Patients' previous adherence to therapy and drugs genetic barriers, not only the number of pills or doses involved, should be taken into consideration in the decision to simplify highly active antiretroviral therapy. PMID- 15127347 TI - Antiretroviral drug content in products from developing countries. AB - Generic and brand name antiretroviral drugs are becoming increasingly available in developing countries. We analyzed 6 antiretroviral medications from 4 international sources for drug content. The active ingredient in tested drug products was within 15% of the labeled amount (range, -12% to +15%) for drugs that were properly stored. PMID- 15127348 TI - Unintended smallpox vaccination of HIV-1-infected individuals in the United States military. AB - We identified 10 individuals who had undiagnosed human immunodeficiency virus type 1 (HIV-1) infection at the time of smallpox vaccination. Mean CD4 cell count was 483 cells/mm3 (range, 286-751 cells/mm3), and mean log10 plasma HIV-1 RNA load was 4.13 copies/cm3 (range, 2.54-5.16 copies/cm3). All vaccinees (3 primary and 7 repeat) had a normal, robust reaction without complications. Smallpox vaccine was well-tolerated in this small series of HIV-1-infected military personnel. PMID- 15127349 TI - Prognostic factors in 61 cases of Staphylococcus aureus prosthetic valve infective endocarditis from the International Collaboration on Endocarditis merged database. AB - Staphylococcus aureus prosthetic valve infective endocarditis (SA-PVIE) is associated with a high mortality rate, but prognostic factors have not been clearly elucidated. The International Collaboration on Endocarditis merged database (ICE-MD) contained 2212 cases of definite infective endocarditis (as defined using the Duke criteria), 61 of which were SA-PVIE. Overall mortality rate was 47.5%, stroke was associated with an increased risk of death, and early valve replacement was not associated with a significant survival benefit in the whole population; however, patients who developed cardiac complications and underwent early valve replacement had the lowest mortality rate (28.6%). PMID- 15127350 TI - Frequency of reduced vancomycin susceptibility and heterogeneous subpopulation in persistent or recurrent methicillin-resistant Staphylococcus aureus bacteremia. AB - Vancomycin susceptibility was checked in isolates from initial and final blood samples obtained from 22 patients with persistent or recurrent methicillin resistant Staphylococcus aureus bacteremia. The minimum inhibitory concentration of vancomycin was determined using Etest and found to have increased in 2 pairs of isolates, and results of screening in 4 mu g vancomycin and a modified population analysis profile suggested heteroresistance in 3 isolates (13.6%). Heteroresistance is not a common cause of persistent or recurrent bacteremia. PMID- 15127351 TI - Respiratory fluoroquinolones: differences in the details. PMID- 15127353 TI - Adverse effects of bacille Calmette-Guerin vaccination in HIV-positive infants. PMID- 15127355 TI - Clinical significance of hepatitis B core antibody positivity in HCV-infected and HCV/HIV coinfected individuals. PMID- 15127356 TI - A multistate outbreak of Salmonella enterica serotype Saintpaul infections linked to mango consumption: a recurrent theme. PMID- 15127357 TI - Persistence of physical symptoms in and abnormal laboratory findings for survivors of severe acute respiratory syndrome. PMID- 15127358 TI - Distribution of hepatitis B virus genotypes in blood donors and chronically infected patients in a tertiary care hospital in southern India. AB - Hepatitis B virus (HBV) genotypes differ in their potential for causing disease. Consecutive patients with chronic HBV infection (CHBV) (n=122) and blood donors (n=67) positive for hepatitis B surface antigen and HBV DNA were genotyped using polymerase chain reaction-restriction fragment-length polymorphism. The ratio of male to female subjects was significantly higher in the blood donor group than in the group of patients with CHBV (P=.0004). Among patients with CHBV, genotype D was detected in 57.3%, genotype A was detected in 18%, and genotype C was detected in 11.5%. Only genotypes D and A were detected in blood donors. The difference between the detection rate of genotype C in patients with CHBV and in blood donors was significant (11.5% vs. 0%; P=.009). Patients with CHBV who had genotype C had higher alanine transaminase (ALT) levels than those who had genotype A (P=.044) or genotype D (P=.014). Detection of genotype C in patients with CHBV and the association of genotype C with higher ALT levels may predict that this genotype has a greater potential for causing disease than other genotypes. PMID- 15127359 TI - Outbreak of necrotizing fasciitis due to Clostridium sordellii among black-tar heroin users. AB - In California, black tar heroin (BTH) use among injection drug users (IDUs) has resulted in an increased number of cases of wound botulism due to Clostridium botulinum, tetanus due to Clostridium tetani, and necrotizing soft-tissue infections due to a variety of clostridia. From December 1999 to April 2000, nine IDUs in Ventura County, California, developed necrotizing fasciitis; 4 died. Cultures of wound specimens from 6 case patients yielded Clostridium sordellii. Some of the patients appeared to have the toxic shock syndrome previously reported to be characteristic of toxin-mediated C. sordellii infection, which is characterized by hypotension, marked leukocytosis, and hemoconcentration. The suspected source of this outbreak was contaminated BTH that was injected subcutaneously or intramuscularly ("skin popped"). This outbreak of C. sordellii infection serves as another example of how BTH can potentially serve as a vehicle for transmitting severe and often deadly clostridial infections, and reinforces the need to educate IDUs and clinicians about the risks associated with skin popping of BTH. PMID- 15127360 TI - Methicillin-resistant Staphylococcus aureus infection in the Texas prison system. AB - Recent reports indicate that correctional facility inmates may be at elevated risk for contracting methicillin-resistant Staphylococcus aureus (MRSA) infection because of overcrowding, poor hygiene, and high rates of diseases causing immunosuppression. The present study of 299,179 Texas inmates who were incarcerated between 1999-2001 indicated an incidence of 12 MRSA infections/1000 person-years. Inmates with circulatory disease, cardiovascular disease, diabetes, end-stage liver disease, end-stage renal disease, human immunodeficiency virus infection or acquired immunodeficiency syndrome, and skin diseases all exhibited elevated rates of MRSA infection. PMID- 15127361 TI - A pediatric cluster of Shigella dysenteriae serotype 1 diarrhea with hemolytic uremic syndrome in 2 families from France. AB - We report a cluster of pediatric diarrhea due to Shigella dysenteriae serotype 1 involving 11 children in France, including the index case, who had returned from Senegal. Child-to-child transmission was documented by ribotyping. Five children developed hemolytic uremic syndrome (HUS). On the basis of our findings, the choice of antimicrobial treatment for infections with S. dysenteriae serotype 1 should take into account widespread drug resistance and the risk of HUS. PMID- 15127362 TI - Comparative genomic hybridization-array analysis enhances the detection of aneuploidies and submicroscopic imbalances in spontaneous miscarriages. AB - Miscarriage is a condition that affects 10%-15% of all clinically recognized pregnancies, most of which occur in the first trimester. Approximately 50% of first-trimester miscarriages result from fetal chromosome abnormalities. Currently, G-banded chromosome analysis is used to determine if large-scale genetic imbalances are the cause of these pregnancy losses. This technique relies on the culture of cells derived from the fetus, a technique that has many limitations, including a high rate of culture failure, maternal overgrowth of fetal cells, and poor chromosome morphology. Comparative genomic hybridization (CGH)-array analysis is a powerful new molecular cytogenetic technique that allows genomewide analysis of DNA copy number. By hybridizing patient DNA and normal reference DNA to arrays of genomic clones, unbalanced gains or losses of genetic material across the genome can be detected. In this study, 41 product-of conception (POC) samples, which were previously analyzed by G-banding, were tested using CGH arrays to determine not only if the array could identify all reported abnormalities, but also whether any previously undetected genomic imbalances would be discovered. The array methodology detected all abnormalities as reported by G-banding analysis and revealed new abnormalities in 4/41 (9.8%) cases. Of those, one trisomy 21 POC was also mosaic for trisomy 20, one had a duplication of the 10q telomere region, one had an interstitial deletion of chromosome 9p, and the fourth had an interstitial duplication of the Prader Willi/Angelman syndrome region on chromosome 15q, which, if maternally inherited, has been implicated in autism. This retrospective study demonstrates that the DNA based CGH-array technology overcomes many of the limitations of routine cytogenetic analysis of POC samples while enhancing the detection of fetal chromosome aberrations. PMID- 15127364 TI - The continuing challenge of lower respiratory tract infections. PMID- 15127363 TI - Somatic and germline instability of the ATTCT repeat in spinocerebellar ataxia type 10. AB - Spinocerebellar ataxia type 10 (SCA10) is an autosomal dominant disorder characterized by ataxia, seizures, and anticipation. It is caused by an expanded ATTCT pentanucleotide repeat in intron 9 of a novel gene, designated "SCA10." The ATTCT expansion in SCA10 represents a novel class of microsatellite repeat and is one of the largest found to cause human diseases. The expanded ATTCT repeat is unstably transmitted from generation to generation, and an inverse correlation has been observed between size of repeat and age at onset. In this multifamily study, we investigated the intergenerational instability, somatic and germline mosaicism, and age-dependent repeat-size changes of the expanded ATTCT repeat. Our results showed that (1) the expanded ATTCT repeats are highly unstable when paternally transmitted, whereas maternal transmission resulted in significantly smaller changes in repeat size; (2) blood leukocytes, lymphoblastoid cells, buccal cells, and sperm have a variable degree of mosaicism in ATTCT expansion; (3) the length of the expanded repeat was not observed to change in individuals over a 5-year period; and (4) clinically determined anticipation is sometimes associated with intergenerational contraction rather than expansion of the ATTCT repeat. PMID- 15127365 TI - Pneumococcal resistance to macrolides, lincosamides, ketolides, and streptogramin B agents: molecular mechanisms and resistance phenotypes. AB - The macrolides, lincosamides, ketolides, and streptogramin B agents (the MLKS(B) antimicrobial agents) have related chemical structures and share similar molecular targets on the 50S ribosomal subunit of Streptococcus pneumoniae. Mutations in rRNA or ribosomal proteins generate a variety of resistance phenotypes. The M phenotype of S. pneumoniae, which predominates in North America, affords low-level resistance to macrolides only (excluding macrolides with 16-member rings) by means of an efflux pump encoded by the mefA gene. The MLS(B) phenotype, which predominates in Europe, affords high-level resistance to macrolides, lincosamides, and streptogramin B agents and arises, in most cases, from dimethylation of adenine 2058 in the 23S rRNA of the 50S ribosomal subunit. Other, less common, phenotypes arise from other 23S rRNA modifications (ML and K phenotypes) or from amino acid substitution (MS(B) phenotype) or insertion (MKS(B) phenotype) into the 50S subunit ribosomal protein L4. In all cases, the decrease in susceptibility to ketolides (for example, telithromycin) is less than the decrease in susceptibility for other MLKS(B) agents. PMID- 15127366 TI - Monotherapy versus dual therapy for community-acquired pneumonia in hospitalized patients. AB - Several medical-specialty professional societies have suggested that combination therapy with a beta -lactam plus a macrolide or doxycycline or monotherapy with a "respiratory quinolone" (i.e., levofloxacin, gatifloxacin, moxifloxacin, or gemifloxacin) are optimal first-line therapy for patients hospitalized with community-acquired pneumonia. These recommendations are based predominantly on retrospective studies that suggest improved rates of morbidity and mortality and hospital length of stay among patients treated in such a fashion. Well-designed, prospective, randomized studies confirming this tenet of therapy have not been published, although numerous prospective studies have provided indirect confirmation. The biological rationale for such a differential response (i.e., favoring combination therapy or fluoroquinolone therapy) includes the immunomodulatory effects of macrolides or more-optimal treatment of primary infection or coinfection with atypical pathogens. Well-designed, prospective, randomized trials are required to best define the effectiveness of combination therapy with a beta -lactam plus macrolide or doxycycline or with a respiratory quinolone in hospitalized patients with community-acquired pneumonia. PMID- 15127367 TI - "Collateral damage" from cephalosporin or quinolone antibiotic therapy. AB - "Collateral damage" is a term used to refer to ecological adverse effects of antibiotic therapy; namely, the selection of drug-resistant organisms and the unwanted development of colonization or infection with multidrug-resistant organisms. The risk of such damage can be assessed for different antibiotic classes by a variety of epidemiologic studies. Cephalosporin use has been linked to subsequent infection with vancomycin-resistant enterococci, extended-spectrum beta-lactamase-producing Klebsiella pneumoniae, beta-lactam-resistant Acinetobacter species, and Clostridium difficile. Quinolone use has been linked to infection with methicillin-resistant Staphylococcus aureus and with increasing quinolone resistance in gram-negative bacilli, such as Pseudomonas aeruginosa. Neither third-generation cephalosporins nor quinolones appear suitable for sustained use in hospitals as "workhorse" antibiotic therapy. PMID- 15127368 TI - Penicillin and macrolide resistance in pneumococcal pneumonia: does in vitro resistance affect clinical outcomes? AB - In vitro resistance to antimicrobial agents is escalating among pathogens responsible for the most serious respiratory tract infections. Some reports have suggested that this has direct clinical implications. Because of penicillin and macrolide resistance in Streptococcus pneumoniae, current guidelines for the initial treatment of respiratory tract infections advocate less reliance on the use of either of these classes of drugs in single-agent therapy. Recent studies that have assessed the impact of beta -lactam and macrolide resistance on clinical outcomes in community-acquired pneumonia fail to provide incontrovertible evidence for a direct link between in vitro resistance and treatment failure. However, there are anecdotal reports of breakthrough bacteremia due to macrolide-resistant pneumococci among patients receiving macrolide therapy, unlike the situation for beta -lactams and penicillin resistant pneumococci. Continued efforts, including in vitro surveillance, appropriate antibiotic use campaigns, and immunization programs, will be important in limiting the spread of drug-resistant S. pneumoniae. PMID- 15127369 TI - Quinolone resistance mechanisms in pneumococci. AB - Quinolones are widely used in the treatment of respiratory infections, in large part because of their activity against Streptococcus pneumoniae and other commonly encountered respiratory tract pathogens. Pneumococcal isolates that are resistant to these "respiratory quinolones" have now begun to emerge. Resistance is attributable to mutations affecting the intracellular targets of these drugs, topoisomerase IV and DNA gyrase; drug efflux contributes to quinolone resistance in some isolates. Most commonly, strains fully resistant to the newer quinolones have one or more mutations affecting DNA gyrase and topoisomerase IV. Although various agents of this class exhibit selectivity in primarily targeting one or the other of these enzymes, the passage of isolates in the presence of any agent can result in selection of mutations affecting both enzymes. Quinolone resistance in S. pneumoniae has arisen in heterogeneous genetic backgrounds but, ominously, has now appeared in strains that are well adapted for regional and global transmission. PMID- 15127370 TI - Quinolone resistance among pneumococci: therapeutic and diagnostic implications. AB - Fluoroquinolones are widely recommended as empirical monotherapy for community acquired pneumonia. Since 1999, case reports of failure of levofloxacin therapy due to levofloxacin-resistant strains of Streptococcus pneumoniae have started to appear. Most worrying is that, in some cases, levofloxacin resistance has been acquired by pneumococci within days of the initiation of therapy. Because use of current clinical antimicrobial resistance breakpoints fail to identify the majority of S. pneumoniae isolates with only first-step mutations, current treatment guidelines not only may have implications with regard to the ability of surveillance programs to detect emerging resistance but may have therapeutic implications as well. PMID- 15127371 TI - Antibiotic resistance in Streptococcus pneumoniae: what does the future hold? AB - The recent emergence of strains of drug-resistant Streptococcus pneumoniae (DRSP) is a serious clinical and public health problem. Several interventions have been proposed to limit the further emergence and spread of DRSP, including campaigns for appropriate antibiotic use and the introduction of pneumococcal conjugate vaccines. Whether the current epidemic of drug resistance in S. pneumoniae is sustainable or will succumb to current efforts to limit its spread will be decided by an interaction of factors related to the pathogen (i.e., the relative fitness of the resistant strains), to the prescription of antibiotic treatment (i.e., changes in selection pressure), and to the host (i.e., the ability to slow the transmission of DRSP). Much investigation is still needed to better ascertain how maintenance of DRSP strains in the community at large is influenced by each factor and affected by current interventions that are based on these factors. PMID- 15127372 TI - Algorithm for differentiation of left and right posterior paraseptal accessory pathway. AB - We studied 196 consecutive patients with posterior paraseptal accessory pathway (AP); 124 showed manifest preexcitation and 72 were concealed AP. Successful ablation was obtained from left-sided approach in 134 patients (left posterior pasaseptal [LPS] group) and from right sided approach in 62 patients (right posterior paraseptal [RPS] group). A ventriculo-atrial (VA) interval of <50 ms recorded at LPS region (VA(LPS)) during right ventricular pacing identified 95 of the 134 patients (71%) with LPS AP with 100% specificity and positive predictive value. In the 101 patients with VA(LPS) >/=50 ms, a difference in VA interval of <20 ms recorded at the His bundle region and LPS region, DeltaVA(H-LPS), during right ventricular pacing predicted RPS AP with a sensitivity of 97%, a specificity of 85% and a positive predictive value of 91%. When these 2 parameters were used together for prediction of LPS or RPS AP, the sensitivity, specificity, and positive predictive value were 96%, 97%, and 98% for LPS AP, and 97%, 96%, and 91% for RPS AP, respectively. This simple new algorithm using VA(LPS) and DeltaVA (H-LPS) during right ventricular pacing successfully discriminates LPS and RPS AP with high sensitivity, specificity, and positive predictive value and could facilitate radiofrequency ablation in patients with posterior paraseptal AP. PMID- 15127373 TI - Intracardiac electrocardiography via a "saline-filled central venous catheter electrocardiographic lead": a historical perspective. AB - The author describes his experience with a "saline-filled central venous catheter electrocardiographic lead" for the recording of intracardiac electrocardiograms provides a brief description of the methodology, refers to this modality's clinical usefulness, furnishes 2 examples illustrating the contribution of the method to clinical diagnosis, and outlines his literature search to find the discoverer/originator of the employment of a saline-filled intracardiac catheter as an electrocardiogram recording lead. PMID- 15127374 TI - Spectral and correlation analyses of the verapamil-induced conversion of ventricular fibrillation to tachycardia in isolated rat hearts. AB - Ventricular tachycardia (VT) is considered to be the most common precursor of ventricular fibrillation (VF). However, the mechanisms underlying the transition from VT to VF remain unclear. Here, we investigated whether and how perfusion of the heart with verapamil, a blocker of L-type calcium channels, changed the macro dynamics of the heart between VT and VF. The experiments were performed with Langendorff perfused isolated rat hearts, in which left ventricular pressure and left ventricular cardiomyogram were measured. Sustained VT or VF was induced by burst pacing of the left ventricular muscles. During sustained VF, verapamil perfusion resulted in the conversion of VF to VT. A cross-correlation analysis between left ventricular cardiomyogram and left ventricular pressure revealed that the correlation coefficient was small during VF, but became larger during VT. This study showed that inactivation of L-type Ca(2+) channels occurred during verapamil-induced conversion of pacing-induced sustained VF to VT, and characterized the changes in macro-dynamics of the heart associated with the transition. PMID- 15127375 TI - Brugada syndrome with atypical ECG: downsloping ST-segment elevation in inferior leads. AB - We present an unusual case of a young Thai immigrant, symptomatic, who had suffered prior episodes of syncope with strong family background: male, first degree relatives, younger than 45 years old who had died suddenly. The rest ECG, with the patient asymptomatic at the time, showed persistent ST-segment elevation, in inferior leads and "mirror" image in the anterior wall, which were not modified with sublingual nitrates, in absence of demonstrable structural heart disease by chest X-rays and echocardiogram, hypothermia, ischemia, or electrolytic disorders. Holter monitoring revealed at dawn, a short episode of polymorphic ventricular tachycardia of short onset extrasystole coupling, which evolved into asystole and sudden cardiac death. We believe this is a sudden unexplained death syndrome, although we did not have a chance to conduct a genetic study. PMID- 15127376 TI - Electrocardiographic "lambda" wave and primary idiopathic cardiac asystole: a new clinical syndrome? PMID- 15127377 TI - Idiopathic Brugada-type electrocardiographic pattern in an octogenarian. AB - The prognosis of idiopathic Brugada-type ECG pattern in asymptomatic people is unknown. We report a case of an 85-year-old man who had persistent Brugada-type ECG pattern without associated clinical symptoms. This illustrates that the persistent Brugada-type ECG can be present with normal longevity. PMID- 15127378 TI - Abnormal QT interval variability in patients with hypertrophic cardiomyopathy: can syncope be predicted? AB - We sought to determine QT variability pattern in patients with hypertrophic cardiomyopathy (HCM) and its relationship with the risk of syncope. QT interval variability was assessed from 24-hour Holter monitoring in 10 HCM patients with history of syncope, 10 HCM patients without history of syncope, and 10 healthy subjects. QT variability was higher in patients with HCM, in particular in those with history of syncope, than in healthy controls. Time domain QT variability did not vary between waking and sleeping hours in HCM patients, whereas it was significantly shorter while asleep in the control group. Increased QT SDANN identified HCM patients with history of syncope with an accuracy of 75%. Our data show that QT variability is abnormal in HCM patients and indirectly support the concept that arrhythmia-related syncope in these patients may be, at least in part, related to an altered control of repolarization. PMID- 15127379 TI - Exercise-triggered transient R-wave enhancement and ST-segment elevation in II, III, and aVF ECG leads: a testament to the "plasticity" of the QRS complex during ischemia. AB - We describe a patient with coronary artery disease who showed transiently augmented R-waves in his electrocardiogram (ECG) during the course of an exercise treadmill test (ETT), an ECG pattern occasionally associated with the hyperacute phase of myocardial infarction and variant angina. This change in the R-waves was noted in II, III, and aVF ECG leads and was associated with ST-segment elevation; both changed gradually and were normalized during the recovery period. Cardiac enzymes after ETT were negative, and arteriography revealed 3-vessel coronary artery disease, with a completely occluded right coronary artery. The ventriculogram showed very mild hypokinesis of the inferior left ventricular wall, while the global ejection fraction was 75%. These ECG changes, noted previously during ETT in precordial ECG leads, are herein reported to occur also in II, III, and aVF ECG leads. The generation of these ECG changes, which hinges upon a late unopposed depolarization occurring in the course and at the site of severe ischemic injury, constitutes a transient focal ventricular conduction abnormality. PMID- 15127380 TI - An evaluation of postoperative P-wave variables after cardiothoracic surgery. AB - Postoperative atrial fibrillation is common after cardiac surgery. Prediction of which patients will develop postoperative atrial fibrillation would be clinically useful. Increased P-wave duration, suggesting atrial conduction delay and measured from preoperative electrocardiograms, predicts postoperative atrial fibrillation. However, postoperative P-wave duration has not been evaluated after cardiac surgery. In this study, we evaluated postoperative P-wave variables (maximum P-wave duration and P-wave dispersion) over 5 days in cardiac surgery patients receiving amiodarone, pacing or no atrial fibrillation prophylaxis. P wave variables gradually shortened as time passed from surgery. Amiodarone did not shorten P-wave measurements throughout therapy, while pacing shortened P waves in the immediate postoperative period; however, shortening was not sustained. P-waves did not differ between those who did and did not develop atrial fibrillation with amiodarone or pacing. Our findings suggest that atrial conduction delay resulting from cardiothoracic surgery tends to resolve over time and may not play a critical role in the etiology of postoperative atrial fibrillation. PMID- 15127381 TI - Spontaneous cardiac resynchronization. AB - A 73-year-old woman with dilated cardiomyopathy presented with heart failure. The ECG showed sinus rhythm with left bundle branch block, left-axis deviation and prolonged QRS duration and frequent ventricular premature complexes from the left ventricular septal wall were present. Ventricular premature beats had narrower QRS duration than sinus node beats conducted through the His-purkinje fibers consistent with resynchronizing beats. The mechanisms of narrowing of the QRS complex produced by premature beats in cases of impaired intra and interventricular conduction are discussed. PMID- 15127382 TI - The nonspecificity of ST-segment elevation > or =5.0 mm in V1-V3 in the diagnosis of acute myocardial infarction in the presence of ventricular paced rhythm. AB - The criterion of ST-segment elevation (+ST) > or=5.0 mm in leads V1-V3 for the diagnosis of acute myocardial infarction (AMI) in patients with paced ventricular rhythm lacks specificity, since it is also encountered in patients with pacemakers and QRS complexes of large amplitudes in leads V1-V3 but without an AMI. This report is based on 2 such patients with electronically-paced rhythms and excessive +ST who did not have an AMI although this diagnosis would have been made if proposed criteria had been strictly employed. Consequently, it is recommended that in such instances the amplitudes of QRS complexes are taken into account when ST-segment elevations in leads V1-V3 are used in the diagnostic algorithm of AMI. PMID- 15127383 TI - Protein-coated poly(L-lactic acid) fibers provide a substrate for differentiation of human skeletal muscle cells. AB - Tissue engineering represents a potential method for repairing damaged skeletal muscle tissue. Extracellular matrix (ECM) proteins were evaluated for their ability to aid in cell attachment, whereas a poly(L-lactic acid) (PLLA) fiber scaffold was tested as a substrate for the differentiation of human skeletal muscle cells. In comparison to uncoated or gelatin-coated PLLA films, cell attachment increased significantly (p < 0.001) on PLLA films coated with ECM gel, fibronectin, or laminin. Myoblasts differentiated into multinucleated myofibers on ECM gel-coated PLLA fibers, and expressed muscle markers such as myosin and alpha-actinin. Oligonucleotide microarray analysis showed similar gene expression profiles for human skeletal muscle cells on ECM gel-coated PLLA fibers as to that observed for myofibers on tissue culture plates. Therefore, PLLA fibers coated with ECM proteins provide a scaffold for the development of skeletal muscle tissue for tissue engineering and cell transplantation applications. PMID- 15127384 TI - Repair of calvarial defects in rats by prefabricated hydroxyapatite cement implants. AB - The aim of the present study was to test the hypothesis that calvarial defects can be repaired by using preformed implants of calcium phosphate bone cement (CPBC) in rats. Sixty adult female Sprague-Dawley rats received full-thickness calvarial nonhealing defects with a diameter of 8 mm. Three different CPBCs were used: group 1: tetracalcium phosphate-based powder; group 2: a blend of amorphous and crystalline calcium phosphate precursors; and group 3: an alpha-tricalcium phosphate (alpha-TCP)-based powder. Implants were left to cure for 25-40 min at room temperature in a silicon mold of 7.9 mm and inserted press fit into the defects. Fifteen animals served as unfilled controls. After 13, 26, and 52 weeks, the material was analyzed qualitatively by using surface-stained undecalcified thick-section specimens and quantitatively by using semiautomated histometry. Kruskal-Wallis tests were applied to compare mean values of periimplant bone formation at a significance level of p < 0.05. Three implants of group 1 fractured during insertion. Resorption of CPBC without complementary bone formation was noticed in these implants. Unfractured implants were resorbed with simultaneous apposition of bone on the implant surface. After 52 weeks, the resorption rate varied between 23.1 and 39.3%. Periimplant bone formation increased continuously on average around all implant types, but it reached statistical significance only in group 2. The results showed that repair of calvarial defects can be achieved by preformed CPBC implants. The rate of resorption of preformed implants is, however, much lower than that reported for in vivo cured CPBC. PMID- 15127385 TI - Micropatterned surfaces prepared using a liquid crystal projector-modified photopolymerization device and microfluidics. AB - A commercial liquid crystal device projector was modified for photopolymerization using its on-board intense light source and a precision optical control circuit. This device projects reduced images generated by a typical personal computer onto the stage where photopolymerization on a surface occurs. This all-in-one device does not require expensive photomasks and external light sources. However, light scattering and diffraction through glass substrates resulted in undesired reactions in areas corresponding to masked (black) domains in mask patterns, limiting pattern resolution. To overcome this shortcoming, two-step surface patterning was developed. First, three-dimensional microstructures of crosslinked silicone elastomer were fabricated with this device and adhered onto silanized glass substrate surfaces, forming microchannels in patterns on the glass support. Then, acrylamide monomer solution containing photoreactive initiator was flowed into these micromold channels and reacted in situ. The resultant polyacrylamide layer was highly hydrophilic and repelled protein adsorption. Cell seeding on these patterns in serum-supplemented culture medium produced cells selectively adhered to different patterns: cells attached and spread only on unpolymerized silanized glass surfaces, not on the photopolymerized acrylamide surfaces. This technique should prove useful for inexpensive, rapid prototyping of surface micropatterns from polymer materials. PMID- 15127386 TI - Effect of fibrous capsule formation on doxorubicin distribution in radiofrequency ablated rat livers. AB - In this study, we report the histological findings of a combined therapy using radiofrequency ablation and intratumoral drug delivery in rat livers, with special attention to wound-healing processes and their effects on drug transport in post-ablated tissue. Doxorubicin-loaded millirods were implanted in rat livers that had undergone medial lobe ablation. Millirods and liver samples were retrieved upon animal sacrifice at time points ranging from 1 h to 8 days. Results demonstrate a clearly defined area of coagulative necrosis within the ablation boundary. The wound-healing response, complete with the appearance of inflammatory cells, neovascularization, and the formation of a fibrous capsule, was also observed. At the 8-day time point, fluorescence imaging analysis showed a higher concentration of doxorubicin localized within the ablation region, with its distribution hampered primarily by fibrous capsule formation at the boundary. Given the variant nature of ablated liver, a mathematical model devised previously by our laboratory describes the data well up to 4 days, but loses reliability at 8 days. These results provide useful mechanistic insights into the wound-healing response after radiofrequency ablation and polymer millirod implantation, as well as the impact this natural corollary has on drug distribution. PMID- 15127387 TI - Poly(carbonate urethane) and poly(ether urethane) biodegradation: in vivo studies. AB - Several strategies have been used to increase the biostability of medical-grade polyurethanes while maintaining biocompatibility and mechanical properties. One approach is to chemically modify or replace the susceptible soft segment. Currently, poly(carbonate urethanes) (PCUs) are being evaluated as a replacement of poly(ether urethanes) (PEUs) in medical devices because of the increased oxidative stability of the polycarbonate soft segment. Preliminary in vivo and in vitro studies have reported improved biostability of PCUs over PEUs. Although several studies have reported evidence of in vitro degradation of these new polyurethanes, there has been no evidence of significant in vivo degradation that validates a degradation mechanism. In this study, the effect of soft segment chemistry on the phase morphology, mechanical properties, and in vivo response of commercial-grade PEU and PCU elastomers was examined. Results from dynamic mechanical testing and infrared spectroscopy suggested that the phase separation was better in PCU as compared with PEU. In addition, the higher modulus and reduced ultimate elongation of PCU was attributed to the reduced flexibility of the polycarbonate soft segment. Following material characterization, the in vivo biostability and biocompatibility of PEU and PCU were studied using a subcutaneous cage implant protocol. The results from the cage implant study and cell culture experiments indicated that monocytes adhere, differentiate, and fuse to form foreign body giant cells on both polyurethanes. It is now generally accepted that the reactive oxygen species released by these adherent macrophages and foreign body giant cells initiate PEU biodegradation. Attenuated total reflectance-Fourier transform infrared analysis of explanted samples provided evidence of chain scission and crosslinking in both polyurethanes. This indicated that the PCU was also susceptible to biodegradation by agents released from adherent cells. These results reinforce the need to evaluate and understand the biodegradation mechanisms of PCUs. PMID- 15127388 TI - Degradation and cell culture studies on block copolymers prepared by ring opening polymerization of epsilon-caprolactone in the presence of poly(ethylene glycol). AB - Poly(epsilon-caprolactone) (PCL) and its block copolymers with poly(ethylene glycol) (PEG) were prepared by ring-opening polymerization of epsilon caprolactone in the presence of ethylene glycol or PEG, using zinc metal as catalyst. The resulting polymers were characterized by various analytical techniques such as (1)H NMR, SEC, DSC, IR, X-ray, ESEM, and CZE. PCL/PEG copolymers with long PCL chains presented the same crystalline structure as PCL homopolymer, whereas PEG-bearing short PCL blocks retained the crystalline structure of PEG and exhibited an amphiphilic behavior in aqueous solutions. Degradation of PCL and PCL/PEG diblock and triblock copolymers was realized in a 0.13 M, pH 7.4 phosphate buffer at 37 degrees C. The results indicated that the copolymers exhibited higher hydrophilicity and degradability compared with the PCL homopolymer. Large amounts of PEG were released from the bulk after 60 weeks' degradation. In vitro cell culture studies were conducted on scaffolds manufactured via solid free form fabrication by using primary human and rat bone marrow derived stromal cells (hMSC, rMSC). Light, scanning electron, and confocal laser microscopy, as well as immunocytochemistry, showed cell attachment, proliferation, and extracellular matrix production on the surface, as well as inside the scaffold architecture. Copolymers showed better performance in the cell culture studies than the PCL homopolymer. PMID- 15127389 TI - Vacuum ultraviolet treatment of polyethylene to change surface properties and characteristics of protein adsorption. AB - The effects of vacuum ultraviolet (VUV) treatment on surface chemical composition morphology and albumin adsorption for low-density polyethylene (LDPE) and high density polyethylene (HDPE) were investigated. The attenuated total reflection Fourier transform infrared (ATR-FTIR) spectra and contact angle measurements indicated the formation of oxygen-containing polar groups and double bonds under VUV photooxidation in the presence of air or under VUV irradiation in vacuum. Scanning electron microscopy revealed the development of regular structure with the period about 1 microm on the surface of LDPE and HDPE during VUV photooxidation. The correlation between amount of tightly adsorbed albumin and surface concentration of carboxyl groups generated by VUV irradiation was found. The aging effect for protein adsorption during long storage of VUV irradiated samples in air or phosphate-buffered saline (PBS) was studied. The obtained results prove the VUV irradiation provides a high potential to regulate protein adsorption on polymers for biomedical applications. PMID- 15127390 TI - Endothelial cell functions in vitro cultured on poly(L-lactic acid) membranes modified with different methods. AB - We recently developed several methods to enhance the cell-polymer interactions. Optimal conditions for each method have been revealed separately by in vitro cell culture. As a practical consideration for construction of tissue-engineered organs, it is necessary to consider which is the most suitable and convenient in clinical applications. To compare the efficiency of these methods with respect to cell functions, poly-L-lactic acid (PLLA) was selected as matrix being modified by 1) aminolysis (PLLA-NH(2)), 2) collagen immobilization with GA (PLLA-GA-Col), 3) chondroitin sulfate (CS)/collagen layer-by-layer (LBL) assembly (PLLA-CS/Col), 4) photo-induced grafting copolymerization of hydrophilic methacrylic acid (MAA) (PLLA-g-PMAA), and 5) further immobilization of collagen with 1-ethyl-3-(3 dimethylamino propyl) carbodiimide hydrochloride (EDAC) (PLLA-g-PMAA-Col). The surface wettability of the modified PLLA was determined by water contact angle measurements. The cell response to the modified PLLA was quantitatively assessed and compared by using human umbilical endothelial cells (HUVECs) culture. Our results indicate that all the modifications can improve the cytocompatibility of PLLA (e.g., cells can attach with spreading morphology, proliferate and secret vWF and 6-keto-PGF(1 alpha)). All the collagen-modified PLLA showed more positive cell response than those purely aminolyzed or PMAA grafted. Among all the methods, collagen immobilization by LBL assembly or GA bridging after aminolysis is more acceptable for the convenience and applicability to scaffolds. PMID- 15127391 TI - Influence of pores created by laser superfinishing on osseointegration of titanium alloy implants. AB - The aim of this study was to assess the osseointegration of copper vapor laser superfinished titanium alloy (Ti6Al4V) implants with pore sizes of 25, 50, and 200 microm in a rabbit intramedullary model. Control implants were prepared by corundum blasting. Each animal received all four different implants in both femora and humeri. Using static and dynamic histomorphometry, the bone-implant interface and the peri-implant bone tissue were examined 3, 6, and 12 weeks postimplantation. Among the laser-superfinished implants, total bone-implant contact was smallest for the 25-microm pores, and was similar for 50- and 200 microm pore sizes at all time points. However, all laser-superfinished surfaces were inferior to corundum-blasted (CB) control implants in terms of bone-implant contact. Within the 12-week study period, remodeling of woven bone initially formed within pores occurred only in the implants with 200-microm pores. Implants with 25-microm pores showed the highest amount of peri-implant bone volume at all time points, indicating that the amount of peri-implant bone was not correlated with the quality of the bone-implant interface. At 3 and 6 weeks postsurgery, we did not find any differences in mineral apposition rates or bone formation rates between the various implant surfaces. However, the peri-implant bone formation rate at the end of the trial was 70 and 62% higher in implants with 50- and 200 microm pores compared with CB implants, respectively. We conclude that, although laser-superfinished implants were not superior to CB control implants in terms of osseointegration, our study has provided further insights into the mechanisms of bone remodeling within pores of various sizes, and may form a basis for future experiments to design optimal implant surfaces with the help of modern laser technology. PMID- 15127392 TI - Bone formation using novel interconnected porous calcium hydroxyapatite ceramic hybridized with cultured marrow stromal stem cells derived from Green rat. AB - The clinical use of cultured marrow stromal stem cells (MSCs) has recently attracted attention in the field of tissue engineering. For the clinical use of the MSCs, a prominent scaffold is needed. A scaffold hybridized with MSCs is transformed into a "bioactive bone substitute," and this provides good osteoconduction. In this study, a novel calcium hydroxyapatite ceramic with an interconnected porous structure (IP-CHA) was used as a scaffold. MSCs were harvested from Green rats containing Green Fluorescent Protein (GFP), and then these hybrids were implanted into the tibias of Sprague-Dawley rats. The purposes of this study were to examine the osteogenic ability of these hybrids without coculture, and to evaluate whether the resulting bone formation originated from the grafted MSCs or the recipient's cells. The hybridized group showed excellent bone formation compared with the IP-CHA-only implant group. Observation of the implanted MSCs revealed that they survived 8 weeks after surgery, and differentiated into osteoblast-like cells, thus providing bone formation. This implantation of the MSCs/IP-CHA composite provides excellent osteoconduction, and is expected to have extensive clinical applications. PMID- 15127394 TI - Vitronectin is significant in the adhesion of lens epithelial cells to PMMA polymers. AB - A major complication of intraocular lens surgery is diminished visual acuity caused by the regrowth of lens epithelial cells (secondary cataract). Polymethylmethacrylate (PMMA) is a commonly used intraocular lens material. This study addresses the mechanisms underlying the initial adhesion of lens epithelial cells to PMMA and a functionalized PMMA-based terpolymer known to inhibit cell proliferation. Rabbit lens epithelial cells were cultured on the test polymer surfaces in medium containing serum depleted of either fibronectin or vitronectin (or both) to identify the role of these proteins in the initial process of cell adhesion. Adherent cells were quantitated after 60 min, and the actin cytoskeleton and focal contact formation were compared in each serum treatment on both polymers. Vitronectin was significantly more effective for initial cell attachment to both polymers than fibronectin. Normal cell spreading on PMMA required vitronectin and was independent of fibronectin, whereas cell spreading on the terpolymer was abnormal and required the presence of fibronectin and vitronectin together. Together, these results help to explain the inhibition of cell proliferation previously shown on the functionalized PMMA. This work contributes to the design of a polymer for use in intraocular lenses that inhibits proliferation of the target cells. PMID- 15127393 TI - Differentiation of preosteoblasts is affected by implant surface microtopographies. AB - Osteogenesis involves the recruitment of multipotent mesenchymal cells and the progressive differentiation of these cells into osteoblasts. The transcription factor Runx2 regulates osteoblast differentiation and expression of genes necessary for the development of a mineralized phenotype. The purpose of this study was to determine if preosteoblast cell differentiation associated with Runx2 and osteocalcin gene expression was influenced by implant surface microtopography. Human embryonic palatal mesenchymal cells (HEPM cells) were cultured on grooved or roughened cpTi implant discs. Cell phenotypes were evaluated with epifluorescent microscopy. Real-time PCR was used for quantitative analysis of Runx2 and osteocalcin gene expression. HEPM cells mineralized when grown on rough and grooved implant surfaces relative to tissue culture plastic. Real-time PCR showed significant (p < 0.05) increases in Runx2 and osteocalcin gene expression in cells cultured on rough and grooved implant microtopographies. These results suggest that preosteoblast cell differentiation is affected by implant surface microtopographies during osseointegration of dental implants. PMID- 15127395 TI - Quantifying the effect of resin type and sterilization method on the degradation of ultrahigh molecular weight polyethylene after 4 years of real-time shelf aging. AB - Alternative sterilization methods including ethylene oxide, gas plasma, and gamma radiation in an inert environment were implemented in the late 1990s, to limit oxidative degradation of ultrahigh molecular weight polyethylene (PE). There was also a simultaneous transition to PE resins that did not contain calcium stearate. Shelf storage duration of PE inserts following gamma-irradiation in air has been correlated to poor clinical performance and increased wear. This study aimed to determine how sterilization method and resin type influenced degradation of PE after 4 years of real-time shelf aging. It was hypothesized that gamma irradiation and stearate containing resins would incur significantly more degradation than nonradiated, stearate-free resins. Gamma-irradiated PE samples in air and nitrogen had a significantly increased density and oxidation index, compared to nonirradiated PE after 4 years of shelf aging. Alternative sterilization methods such as ethylene oxide and gas plasma appeared to have significantly less oxidation regardless of PE resin type. A partial correlation demonstrated that density and oxidation index were not correlated (r(2) = 0.079) when examining the influence of sterilization method. The data supported that after 4 years of real-time shelf aging, the type of sterilization method had a larger influence on PE degradation than resin type. PMID- 15127396 TI - Advanced bioceramic composite for bone tissue engineering: design principles and structure-bioactivity relationship. AB - The synthesis of a new resorbable porous bioactive silica-calcium phosphate composite (SCPC) that can be used as a tissue-engineering scaffold for bone regeneration is described. The effects of chemical composition and thermal treatment on crystallization and the mechanism of phase transformation in SCPC were evaluated. In the silica-rich samples, beta-rhenanite (beta-NaCaPO(4)) and alpha-cristobalite (SiO(2)) were the dominant phases after treatment at 800 degrees C. On the other hand, in the calcium phosphate-rich samples, calcium pyrophosphate (Ca(2)P(2)O(7)) was formed in addition to beta-rhenanite and alpha cristobalaite. X-ray diffraction analyses showed a shift in the 2 theta value of the main peak(s) of all phases indicating the formation of solid solutions. Phase transformation reactions were accompanied by a loss of water molecules that contributed to the formation of pores in the size range 10-300 microm. All SCPC samples adsorbed a significantly higher quantity of serum protein than bioactive glass (p < 0.0001). In addition, the silica-rich SCPC adsorbed a significantly higher amount of serum protein than the calcium phosphate-rich samples (p < 0.003). While the crystallization of amorphous silica into L-quartz significantly inhibited serum protein adsorption, the transformation of L-quartz into alpha cristobalite solid solution (ss) significantly enhanced protein adsorption. On the other hand, in conjunction with the transformation of brushite (CaHPO(4)) into pyro- and tri-calcium phosphates, there was a significant decrease in protein adsorption. However, as pyro- and tri-calcium phosphates transformed into beta-rhenanite, by thermal treatment, protein adsorption increased markedly. Critical-size bone defects grafted with silica-rich SCPC were filled with new bone and contained minimal residues of the graft material. Bone defects grafted with bioactive glass enhanced new bone formation, however, with very limited resorption. The enhanced resorption of SCPC in vivo correlates well with the higher rate of silica dissolution from SCPC than bioactive glass. The facilitated Si dissolution was associated with rapid bone regeneration in defects grafted with SCPC. The enhanced bioactivity properties of the SCPC are due to its chemical composition, modified crystalline structure, and high porosity. The new SCPC may be used for a wide variety of applications in the field of bone reconstruction including tissue-engineering scaffolds for cell and drug delivery. PMID- 15127397 TI - Molecular release from a polymeric microreservoir device: Influence of chemistry, polymer swelling, and loading on device performance. AB - A polymeric microreservoir device for controlled-release drug delivery relies on the degradation of thin poly(lactic-co-glycolic acid) membranes that seal each reservoir to achieve pulsatile drug delivery. In vitro release studies in which the swelling of the reservoir membranes was measured indicate a correlation between the release times of various radiolabeled molecules from the devices and the time at which the maximum membrane swelling was observed. Varying the chemistry (lipophilicity/hydrophilicity) or molecular weight of the molecules loaded into the devices did not appear to affect the degree of membrane swelling that was observed, or the time at which the molecules were released from the devices. The amount of drug that was loaded into the reservoirs also did not appear to affect the observed release time of the drug from the device, a significant departure from the behavior of many matrix-type polymeric drug delivery systems. PMID- 15127398 TI - Molecular responses of vascular smooth muscle cells to paclitaxel-eluting bioresorbable stent materials. AB - We studied the influence of paclitaxel, eluted from poly(L-lactic acid) (PLLA), on cultured vascular smooth muscle cell (VSMC) proliferation as a model of bioresorbable stent-induced restenosis. We blended paclitaxel in cast PLLA films (P-PLLA), demonstrating controlled release of the drug, then studied VSMC adhesion, proliferation, and gene expression profiles. No difference in cell adhesion was found between P-PLLA and PLLA controls (105 +/- 12% of PLLA controls). However, P-PLLA significantly reduced VSMC proliferation (40 +/- 15% of PLLA controls, p < 0.05). Using cDNA microarray technology, we identified major effects of P-PLLA, including: upregulation of genes related to apoptosis, anti-proliferation and antioxidation; and suppression of cell cycle regulators and cell survival markers. The expression patterns indicate that P-PLLA regulates gene expression and cell functions via new pathways, including receptor tyrosine kinase (RTKs), mitogen-activated protein kinase (MAPKs), and protein kinase (PKs, e.g., PKA) pathways, in addition to the stabilization of polymerized microtubules. PMID- 15127399 TI - Structural changes of fibronectin adsorbed to model surfaces probed by fluorescence resonance energy transfer. AB - Structural changes of proteins during adsorption to biomaterials affect the presentation of molecular binding sites and, ultimately, biomaterial performance. We have applied fluorescence resonance energy transfer (FRET) spectroscopy to study structural changes of the cell adhesion protein, fibronectin (Fn), following adsorption to model hydrophilic and hydrophobic surfaces. Fn was labeled with donor and acceptor fluorophores using two labeling schemes and intramolecular energy transfer was calibrated against measured structural changes of Fn in denaturing solutions. FRET was then applied to measure Fn's structure on surfaces. Based on FRET, Fn underwent greater extension of its dimer arms on hydrophilic glass than on hydrophobic fluoroalklysilane-derivatized glass (fluorosilane), and this extension was insensitive to molecular packing over a range of adsorption concentrations. Fn's conformation on glass better promoted cell attachment than on fluorosilane; the roles of both global structural changes (movements of modules) and local structural changes (disruption of secondary structure) on Fn's cell integrin binding activity are discussed. Based on previous FRET work, we compare Fn's conformations on these surfaces with its conformations in fibroblast culture. FRET is unique in allowing direct comparison of protein structure between biomaterial surfaces and cell culture. PMID- 15127400 TI - Modulation of differentiation and mineralization of marrow stromal cells cultured on biomimetic hydrogels modified with Arg-Gly-Asp containing peptides. AB - We synthesized biomimetic hydrogels modified with an osteopontin-derived peptide (ODP) and used them as a substrate for in vitro culture of marrow stromal cells (MSCs) to investigate the effect of the biomimetic surface on differentiation of MSCs into osteoblasts. Proliferation and biological assays for 16 days proved that MSCs became differentiated into osteoblasts secreting osteogenic phenotypic markers such as alkaline phosphatase (ALP), osteopontin, and mineralized calcium. In addition, there was an additive effect of the cell-binding peptide on differentiation and mineralization of MSCs cultured in the presence of soluble osteogenic supplements in cell culture media. For example, calcium content at day 16 on peptide-modified hydrogels was significantly higher than on tissue culture polystyrene. Two general trends were observed: (1) proliferation of MSCs decreased as the amount of differentiation markers increased, and (2) higher peptide concentrations accelerated the differentiation of MSCs. On the hydrogel modified with ODP, ALP activity exhibited a maximum value of 36.7 +/- 4.2 pmol/cell/h at day 10 for the concentration of 2 micromol/g while the culture time needed for maximum ALP activity occurred on day 13 for the lower concentrations. On the same hydrogel, the calcium content at day 10 was 21.4 +/- 2.3 ng/cell for the peptide concentration of 2 micromol/g and 1.0 +/- 0.3 ng/cell for 1.0 micromol/g. We used Gly-Arg-Gly-Asp-Ser (GRGDS) for modification of the hydrogel as a comparison to the results with ODP. However, osteoblast development was not significantly affected by the nature of the binding peptide sequences. These results suggest that MSC function can be modulated by variation of the peptide concentration in biomimetic hydrogels used for scaffold-based bone tissue engineering. PMID- 15127401 TI - Selective protein adsorption and blood compatibility of hydroxy-carbonate apatites. AB - We examined the blood compatibility and protein adsorption on hydroxyapatite and hydroxy-carbonate apatite. Those apatites were synthesized under a 0, 5, or 15% CO(2)-containing N(2) atmosphere by a wet-chemical method with a strong ammonia alkali solution of calcium nitrate and diammonium hydrogen phosphate (5:3 in molar ratio) and subsequent calcination in the range of 105-700 degrees C. From infrared (IR) analysis, the carbonate ions substituted both phosphate ions and hydroxyl ions in the hydroxyapatite lattice; the intensities of IR bands assignable to phosphate ions and hydroxyl ions were reduced on calcinations. The specific surface areas of synthesized apatites decreased with increasing calcination temperature. Blood-clotting properties were evaluated in terms of active partial thromboplastin time, prothrombin time, and the amount of fibrinogen for the plasma in contact with the apatites, indicating that all the apatites barely influenced the blood clotting system. The apatites were in contact with a solution containing both bovine serum albumin (BSA) and beta(2) microglobulin (beta(2)-MG), and the amounts of those proteins adsorbed on them were examined: the amount of absorbed BSA and beta(2)-MG gradually increased with the calcination temperature below 500 degrees C, while it showed a sudden increase when more than 600 degrees C. Hydroxy-carbonate apatite synthesized under a 15% CO(2)-containing N(2) atmosphere and calcined below 400 degrees C had the greatest selectivity in adsorbing beta(2)-MG. Thus, a higher selectivity for beta(2)-MG adsorption was empirically correlated to carbonate ions incorporated in the hydroxyapatite lattice. PMID- 15127402 TI - Effect of controlled zinc release on bone mineral density from injectable Zn containing beta-tricalcium phosphate suspension in zinc-deficient diseased rats. AB - The purpose of this study was to evaluate the efficacy of zinc (Zn)-containing beta-tricalcium phosphate (Zn-TCP) in correcting the bone mineral deficiency noted in osteoporosis using ovariectomized rat model. Four rats were used for each of the four experimental groups: D0, D10, D20, and N10. The rats in D0, D10, and D20 groups were ovariectomized, and fed a vitamin D-, Ca-, and Zn-deficient diet, and induced Zn-deficient osteoporoses for 9 weeks. In contrast, the N10 group was the normal rats fed normal healthy diet for 9 weeks. D0 group was injected with pure beta-TCP suspension, D10 and D20 groups were injected with suspensions containing 10 mg of 10 mol % (6.17 wt % Zn) and 20 mol % (12.05 wt % Zn) Zn-TCP, respectively, and the healthy group, N10 were injected with 10 mol %. Zn-TCP suspensions. Injections were administered intramuscularly in the left thigh once a week in all rats, and fed a vitamin D- and Zn-deficient diet for 9 weeks. The plasma calcium (Ca) and Zn levels, plasma alkaline phosphatase activity (ALP) and bone mineral density (BMD) of the lumbar vertebra and femora were measured. The plasma Zn levels in all the rats were between 1.1 and 2.8 microg/mL. The areas under the curves for the Ca, Zn, and ALP (Ca-AUC, Zn-AUC, and ALP-AUC) levels between 0 and 63 days were calculated. Results for the AUCs were as follows: (1) the Zn-AUCs were in the order of N10 = D20 > D10 > D0; (2) the Ca-AUCs for D0, D10 groups were significantly lower than that for the N10 group; (3) the ALP-AUCs for the D10 and D20 groups were significantly higher than that for the N10 group, and that of the D0 group was in between those. The body weight of D10 and D20 groups significantly increased with time, that of the D0 group increased slightly, and that of the N10 group remained unchanged for the entire experimental period. The BMD of the lumbar vertebrae of the D10 and D20 groups (about 100 mg/cm(2)) was significantly higher than that of the D0 group but lower than that of the N10 group. The BMD of the left femur increased more than that of the right femur with the increase in the amount of Zn in the suspension. The results of this study suggest that the local effect on BMD was more pronounced than the effect on the whole body. PMID- 15127403 TI - Titanium implants enhance pulmonary nitric oxide production and lung injury in rats exposed to endotoxin. AB - An increase in levels of elemental Ti in the blood and lung of rats with a Ti alloy implant has been demonstrated. However, the pathophysiological role of the elevated elemental Ti level in the circulation remains unclear. Rats were implanted with Ti alloy discs for 4 weeks. The levels of elemental Ti in the blood and lung were especially increased compared with other tissues. The Ti alloy implant enhanced lung injury related to endotoxin from Gram-negative bacteria (lipopolysaccharide, LPS), which was characterized by lung edema and other histological changes such as recruitment of neutrophils, interstitial edema, and alveolar hemorrhage in the lung. In the presence of endotoxin, an increase of nitrite production was shown in the plasma and bronchoalveolar lavage fluid of rats implanted with a Ti alloy. Moreover, the Ti alloy implant further enhanced the induction of inducible nitric oxide (NO) synthase (iNOS) protein expression induced by LPS in the lung. These endotoxin-related responses in the presence or absence of the Ti alloy implant could be inhibited by aminoguanidine (an iNOS inhibitor). These results provide the first experimental evidence that circulating Ti released from Ti alloy implants has an ability to affect pulmonary iNOS protein expression, and enhance the pathogenesis of acute lung injury during endotoxemia. PMID- 15127404 TI - Determining relevance of a weight-bearing ovine model for bone ingrowth assessment. AB - A weight-bearing ovine model was used to quantify cancellous bone ingrowth and remodeling in porous-coated implants over 6, 12, and 24 weeks in situ. The null hypothesis for the investigation was that there would be no significant difference between the amount of cancellous bone ingrowth and rate of remodeling in this ovine model compared to a reported human bilateral implant model. Bone ingrowth progressed from 20.1 +/- 8.2% at 6 weeks in situ to 23.8 +/- 7.9% at 12 weeks, and 30 +/- 5.1% at 24 weeks. Fluorochrome analysis demonstrated a mineral apposition rate of 1.07 +/- 0.28 microm/day for bone at the porous-coating interface, whereas host bone remodeling at 0.89 +/- 0.23 microm/day. Histological analysis showed no adverse tissue or inflammatory response. The null hypothesis was supported in that regression analysis demonstrated that the amount of cancellous bone ingrowth over time (p = 0.545) and mineral apposition rate over time (p = 0.089) in this ovine model was not significantly different than reported human bilateral knee data. The results of this study appear to validate the ovine model for use in understanding skeletal attachment of porous-coated implants to cancellous bone in humans. PMID- 15127405 TI - Gentamicin release from two-solution and powder-liquid poly(methyl methacrylate) based bone cements by using novel pH method. AB - The release of gentamicin as a function of time was measured for Palacos and two solution bone cements by using a novel pH technique. The pH of an aqueous solution of gentamicin is a function of the gentamicin concentration and it decreases linearly over concentrations of 0.0-0.1 wt %. Therefore, a new, direct, and inexpensive in vitro technique was developed based on continuous readings of the pH in phosphate-buffered saline (PBS) at 37 degrees C to determine the release kinetics of gentamicin from poly(methyl methacrylate) (PMMA)-based bone cement. In addition, this method was used to compare the release profiles of Palacos R-40 bone cement with a two-solution bone cement developed in our laboratory and loaded with two different concentrations of gentamicin sulfate. Finally, the pH-based method was used to track the elution of gentamicin in both mixed and static conditions to determine the effect of mixing on the diffusion of gentamicin out of the cement. It was found that Palacos R-40 released 4.95 +/- 0.22 wt % of its gentamicin after 24 h in PBS solution. This data compares favorably with previously reported values of gentamicin elution from Palacos R 40, which ranged from 3 to 8 wt % of the total amount of incorporated gentamicin, depending on the size and the surface area of the samples. The results show that Palacos samples released 4.84 +/- 0.27 mg after 24 h, a two-solution cement loaded with an equivalent concentration of gentamicin sulfate released 3.81 +/- 0.52 mg, and two-solution cement loaded with twice the concentration of Palacos released 5.53 +/- 0.26 mg of gentamicin. A higher percentage of release was recorded from Palacos than from the two-solution bone cement, and the effect of PBS mixing conditions on the release kinetics was only significant in the early stages of release and not at 24 h. It was concluded that monitoring the pH is an effective technique to measure gentamicin release from PMMA-based bone cements in PBS solution. PMID- 15127406 TI - The in vivo calcification capacity of a copolymer, based on methacryloyloxyethyl phosphate, does not favor osteoconduction. AB - Polymers can be interesting alternatives to bone grafts; they must present suitable mechanical and osteoconductive properties. Biomimetic properties may be a key factor for the recognition by bone cells. Methacryloyloxyethyl phosphate (MOEP) was found to enhance hydroxyapatite deposition. The copolymer containing MOEP and 1-vinyl-2-pyrrolidinone (50-50%) binds large amounts of calcium. Particles of the copolymer were used to fill large cranial bone defects in the rat. After a 12-week healing period, the animals were euthanized and the skulls examined by X-ray, histology, and electron microscopy (EM). The high phosphate content of the polymer conferred a marked calcium-binding capacity, and the particles were heavily calcified. They were embedded in a light fibrous stroma containing numerous capillaries and multinucleated giant cells. The osteoconductive properties were poor: only few trabeculae developed centripetally from the margins of the defects. There was no bone bonding and no osteoblast on the surface of the calcified material. Backscattered EM revealed that the degree of calcification was homogeneous in all particles. Calcium-phosphorus calcospherites were never observed. The material appeared to trap calcium but to impair nucleation because only small hydroxyapatite tablets were occasionally observed. Polyphosphated materials do not represent a suitable source of potentially usable bone substitutes. PMID- 15127407 TI - The challenges of cancer survivorship. PMID- 15127408 TI - Exercise echocardiography reflects cumulative anthracycline exposure during childhood. PMID- 15127409 TI - Exercise echocardiography reflects cumulative anthracycline exposure during childhood. AB - BACKGROUND: Low and moderate dose anthracycline therapy used in current treatment protocols, may be associated with subclinical abnormalities of cardiac function and late presentation with congestive heart failure. Exercise echocardiography is a useful and non-invasive means of uncovering latent cardiac dysfunction in other settings. The purpose of this study is to determine whether exercise echocardiography has the potential to detect subclinical abnormalities of cardiac function in children treated with anthracyclines. PROCEDURE: One hundred ten children previously treated with anthracycline, in remission and off treatment for >12 months were assessed, together with 31 control subjects. Each subject had a resting ECG and echocardiogram performed, and following exercise on a treadmill according to the Bruce protocol, a repeat echocardiogram was performed. RESULTS: Cumulative anthracycline dose was the only patient variable related to any of the outcome measures. Resting fractional shortening was normal in the majority of treated patients but was inversely related to cumulative anthracycline dose (rate of decline 1.2%/100 mg/m(2)). Following peak exercise, the inverse relationship between fractional shortening and anthracycline dose was more pronounced (rate 2.7%/100 mg/m(2)). CONCLUSIONS: Higher anthracycline doses are associated with a greater difference in peak fractional shortening between treated subjects and controls. Exercise echocardiography is a simple, relatively inexpensive tool that may enhance the detection of latent cardiac dysfunction after anthracycline administration during childhood. PMID- 15127410 TI - Long-term cause-specific mortality among five-year survivors of childhood cancer. AB - BACKGROUND: The purpose of our study was to assess long-term cause-specific mortality of 5-year childhood cancer survivors. PROCEDURE: The study population consisted of 1,378 patients who had been treated for childhood cancer in The Netherlands between 1966 and 1996 and survived at least 5 years; follow-up was complete for 99% of survivors. Cause-specific mortality was compared with general population rates to assess relative and absolute excess risks of death (standardized mortality ratio (SMR) and AER). RESULTS: After a median follow-up of 16.1 years, 120 patients had died. The overall SMR was 17-fold (95% CI: 14.3 20.6) increased compared to the general population. Our cohort experienced an excess of 7 deaths per 1,000 person-years. Patients who received combined modality treatment and were treated for at least one recurrence experienced the highest risk of death (SMR = 92.3; AER = 37.0 per 1,000 person-years). The SMR appeared to stabilize at an about 4 to 5-fold increased risk of death after 20 years of follow-up. Only after more than 20 years of follow-up excess mortality due to other causes than the primary cancer exceeded mortality from the primary childhood cancer (2.3 vs. 0.3/1,000 patients/year). The SMR for all causes other than primary cancer was 5.4 in 25-year survivors. The overall risks of death strongly decreased with increasing attained age, with an SMR of 1.6 (n.s.) and an AER of 0.3 per 1,000 person-years for survivors of 30 years or older. CONCLUSIONS: The first primary cancer contributes most to the absolute excess risk of death in 5-year survivors of childhood cancer, but after 25 years childhood cancer mortality is negligible. Relative risk of death due to other causes is still significantly increased after 25 years of follow-up. PMID- 15127411 TI - Second neoplasms after treatment of childhood cancer in Slovenia. AB - BACKGROUND: The number of long time survivors of childhood cancer treatment is constantly increasing over the last decades as a result of advances in diagnosis and treatment. The occurrence of second neoplasms is one of most serious late effects observed in cancer survivors. METHODS: The risk of secondary neoplasm was studied in a cohort of 1,577 patients treated for childhood cancer registered in the Cancer Registry of Slovenia (CRS) between 1961 and 2000. The time at risk was defined from the date of diagnosis of first malignancy to the time of death or the end of the study. RESULTS: The most frequent primary malignancies were: acute leukemia 28.5%, central nervous system (CNS) tumors 21.3%, and lymphomas 16.6%. Median observation time was 7.8 years. Forty-eight patients developed second neoplasms. CNS tumors, acute leukemias, and thyroid carcinoma were most frequent second neoplasms. The cumulative risk for second neoplasm in the entire cohort was 0.06% at 5 years, 5.1% at 15 years, and 12.6% at 25 years after diagnosis of first cancer. The overall survival after second neoplasm was 65% 10 years after the diagnosis of second neoplasm. CONCLUSIONS: Patients after treatment of childhood cancer are at special risk for subsequent neoplasms and long-term follow-up is mandatory. PMID- 15127412 TI - Young adult survivors of childhood acute lymphoblastic leukemia: spontaneous GH secretion in relation to CNS radiation. AB - BACKGROUND: Young adults who are long-term survivors of acute lymphoblastic leukaemia (ALL) in early childhood usually do well and do not have to go to regular medical checkups. Many of these survivors did receive prophylactic cranial radiotherapy during their oncological treatment. The effect of cranial irradiation on the hypothalamus is considered to be progressive. Therefore, late effects, such as reduced growth hormone (GH) secretion, may remain undetected until adulthood. PROCEDURE: Records from all patients treated for ALL before the onset of puberty in the region of West Sweden, between 1 January 1973 and 31 December 1985 were included, provided they were in first remission with a minimum follow-up time of 15 years, and a minimum age of 20. These criteria were met by 47 young adults aged 20-32 years, of whom 35 agreed to participate. We studied spontaneous GH secretion over 24 hr, IGF-I and IGFBP-3, final height and BMI. The patients had been treated according to three consecutive Swedish childhood leukaemia group protocols. The median follow-up time was 20 years, and 19 of the patients had been treated with cranial irradiation (CRT+), 16 had not (CRT-). RESULTS: CRT+ patients had significantly lower maximal peaks of GH than CRT- patients. Fifty percent of the CRT+ patients had a GH(max) below the cut-off level (3.3 microg/l), for GH treatment. CRT- patients all had GH(max) levels considered within the normal range. Final height of all the patients, except one CRT+ women, was in the range of expected midparental height, the median loss in final height in the CRT+ patients was 0.8 standard deviation (SD). No patient in this study was obese by definition (BMI <30 kg/m(2)). IGF-I and IGFBP-3 concentrations did not correlate to variations in spontaneous GH secretion in these patients. CONCLUSIONS: In spite of the little effect on final height, we found impaired spontaneous GH secretion in 79% of young adults 20-32 years of age, and GH deficiency (GHD) in 47% after low-dose cranial irradiation in early childhood. The consequences of this low-GH secretion need to be investigated. PMID- 15127413 TI - Fertility in women treated with cranial radiotherapy for childhood acute lymphoblastic leukemia. AB - BACKGROUND: Fertility impairments among women treated during childhood for cancer are known to occur after some, but not all, types of anticancer therapy. Although leukemia is the most common cancer of childhood, until now fertility in survivors has not been comprehensively assessed. PROCEDURE: We investigated functional impairment of fertility in women who were long-term survivors of acute lymphoblastic leukemia (ALL) with a retrospective cohort study. Proven fertility (defined as ever pregnant) was evaluated by self-report among 182 females treated on protocols of the Children's Cancer Group (age at interview, 22.6 years on average) and 170 controls drawn from among the survivors' female siblings (23.4 years). The interview included psychosocial inventories designed to detect mood problems. RESULTS: Significant fertility deficits were noted in female survivors treated with cranial radiotherapy (CRT) at any dose around the time of menarche (relative fertility (RF)) = 0.27, 95% CI = 0.09, 0.82, P = 0.03). Controlling for marital status, mood at interview, and many fertility-related situations did not change the association. CONCLUSION: This study provides evidence for fertility deficits after treatment for ALL with CRT, and, in addition, for the first time, suggests that girls treated around the time of menarche are especially at risk. Clinical confirmation of these results is needed. If gonadal damage occurs in women receiving these treatments, their risk for further sequelae, such as osteoporosis and heart disease, may be significantly raised, requiring active management and intervention. PMID- 15127414 TI - Ototoxicity following pediatric hematopoietic stem cell transplantation: a prospective cohort study. AB - BACKGROUND: Our objectives were to describe the frequency and determine risk factors for hearing deterioration following pediatric stem cell transplantation (SCT). METHODS: In this prospective cohort study, we performed pure tone audiometry and measured otoacoustic emissions (OAE) prior to and following SCT. Worse hearing was considered deterioration in either audiometry or OAE. RESULTS: Between October 2000 and November 2002, 45 informative audiometry or OAE results were obtained. Hearing deteriorated following SCT in 20/45 (44%) of these children. Those with worse hearing following SCT were more likely to have neuroblastoma (odds ratio [OR] 16.0 [95% CI 1.8, 143.2; P = 0.003]), receive carboplatin conditioning (OR 7.7 [95% CI 1.4, 41.9; P = 0.01]), have abnormal baseline hearing (OR 5.1 [95% CI 1.3, 19.5; P = 0.02]), and have higher baseline serum creatinine (OR for every increase of 5 micromol/L of serum creatinine of 1.5 [95% CI 1.03, 2.1; P = 0.03]). CONCLUSION: Many children who undergo SCT will have deterioration in hearing following SCT. A high-risk group of children can be delineated who may benefit from more intensive audiological monitoring following SCT. PMID- 15127415 TI - Posttraumatic stress symptoms in adult survivors of childhood cancer. AB - BACKGROUND: Previous research suggests that posttraumatic stress disorder (PTSD) is present in survivors of childhood cancer. The aim of the current study was to explore posttraumatic stress symptoms in a sample of young adult survivors of childhood cancer. In addition, the impact of demographic, medical and treatment factors on survivors' posttraumatic stress symptoms was studied. PROCEDURE: Participants were 500 long-term survivors of childhood cancer. The median age at follow-up was 24 years (age range, 16- 49 years, 47% female). To assess symptoms of posttraumatic stress, all participants completed the Impact of Event Scale (IES), a self-report instrument consisting of two subscales, intrusion and avoidance. RESULTS: Twelve percent of this sample of adult survivors of childhood cancer had scores in the severe range, indicating they are unable to cope with the impact of their disease and need professional help. Twenty percent of the female survivors had scores in the severe range as compared with 6% of the male survivors. Linear regression models revealed that being female, unemployed, a lower educational level, type of diagnosis and severe late effects/health problems were associated with posttraumatic stress symptoms. CONCLUSIONS: The results indicate that, although the proportion of survivors reporting symptoms is well within the proportions found in the general population, a substantial subset of survivors report symptoms of posttraumatic stress. This finding supports the outcomes reported previously that diagnosis and treatment for childhood cancer may have significant long-term effects, which are manifested in symptoms of posttraumatic stress. The investigated factors could explain posttraumatic stress symptoms only to a limited extent. Further research exploring symptoms of posttraumatic stress in childhood cancer survivors in more detail is clearly warranted. From a clinical perspective, health care providers must pay attention to these symptoms during evaluations in the follow-up clinic. Early identification and treatment of PTSD symptoms can enhance the quality of life for survivors of childhood cancer. PMID- 15127416 TI - MRI morphometric and neuropsychological correlates of long-term memory in survivors of childhood leukemia. AB - BACKGROUND: Long-term survivors of acute lymphoblastic leukemia (ALL) are reported to have post-treatment neurological changes as well as neuropsychological changes. Few studies have investigated the relationship between magnetic resonance imaging (MRI) volumetric measurements of brain regions of interest and cognitive functioning. This study examined the relationship between hippocampal volumes and long-term memory abilities in survivors of ALL. PROCEDURE: Ten survivors of ALL and ten matched controls underwent MRI acquisition. The participants completed a battery of tests measuring both visual and verbal long-term memory. Volumetric measurements of the hippocampus were obtained by consecutive manual tracing using the NIH Image 1.52 program. Estimates of whole brain volume were also obtained. RESULTS: No significant group differences were found in right or left hippocampi. Nor were there significant differences between the two groups on measures of long-term memory. Correlations between volumetric measurements of the hippocampus and measures of long-term memory were non-significant. CONCLUSIONS: Our hypothesis of deficits in both long term memory and its neural substrates was not supported. It was concluded that the hippocampus, as an early developing structure, may be less vulnerable to chemotherapy treatment. Children surviving ALL are able to retain and retrieve once learned information comparable with peers. PMID- 15127417 TI - Results of treatment of fifty-six patients with localized retroperitoneal and pelvic rhabdomyosarcoma: a report from The Intergroup Rhabdomyosarcoma Study-IV, 1991-1997. AB - BACKGROUND: We reviewed 56 IRS-IV patients with localized rhabdomyosarcoma [RMS] of the retroperitoneum/pelvis to assess outcome and prognostic factors, including the value of initially excising >or=50% of the tumor (debulking) before chemotherapy. METHODS: Patients had embryonal RMS [N=38], alveolar RMS [N = 7], RMS not otherwise specified [NOS, N = 7], or undifferentiated sarcoma [N = 4]. Fifteen patients were debulked; 41 patients were biopsied. All received VAC; most received radiotherapy. RESULTS: Estimated 5-year failure-free survival [FFS] and overall survival rates were 70 and 75%, respectively. FFS rates were better for patients <10 years old and those with embryonal RMS compared to alveolar RMS/undifferentiated sarcoma. After adjusting for age and histological differences, FFS was better for patients whose tumor was debulked prior to beginning therapy [P = 0.02]. CONCLUSIONS: These results are superior to those of previous protocols for patients with RMS of the retroperitoneum/pelvis. Initial excision of >or=50% of the tumor may be associated with increased FFS. PMID- 15127418 TI - Extra-target doses in children receiving multileaf collimator (MLC) based intensity modulated radiation therapy (IMRT). AB - PURPOSE: To investigate the extra-target doses using intensity modulated radiation therapy (IMRT). MATERIALS AND METHODS: Thirteen children underwent multileaf collimator (MLC)-based IMRT. Treatment site was head and neck or brain in eight (Group I), trunk in two (Group II), and abdomen/pelvis in three (Group III). Thermoluminescent dosimeters (TLD) were placed at the thyroid gland, breast, and testis. A control group of seven children received conventional RT and TLD measurements. RESULTS: For the eight Group I children, the median dose equivalent measurements during the course of IMRT to the thyroid, breast, and testis were 348 mSv, 110 mSv, and 30 mSv, respectively. For the two Group II patients, the measurements to the thyroid ranged from 1,525 to 2,449 mSv while for the testis was 62 mSv. For the Group III patients, the median dose equivalent measurements to the thyroid, breast, and testis were 182 mSv, 406 mSv, and 159 mSv. The median dose equivalent measurements to the thyroid, breast, and testis for Group I children were 300 mSv, 120 mSv, and 75 mSv. The Group II conventional patient had a measurement of 180 mSv, 80 mSv, and 80 mSv to the thyroid, breast, and testis. For the Group III conventional cases, the median dose equivalent measurements were 192 mSv, 496 mSv, and 434 mSv. CONCLUSIONS: No significant difference was seen in the thyroid and breast doses of children receiving MLC based IMRT compared to conventional RT for the treatment of head and neck/brain and abdominal/pelvic tumors. PMID- 15127419 TI - Childhood blastic NK cell leukemia successfully treated with L-asparagenase and allogeneic bone marrow transplantation. AB - Blastic NK cell lymphoma/leukemia is a rare and highly malignant neoplasia in both adults and children. It is characterized by lymphoblastoid morphology without cytoplasmic granules and immature NK cell immunophenotypes (CD56+, CD57-, CD16-). It has predilection for extranodal organ involvement, and the prognosis of affected patients is extremely poor under the current chemotherapy. We present a 14-year-old girl who was diagnosed as having blastic NK cell leukemia with mediastinal, pleural, and pericardial involvement. Immunophenotyping of her leukemic cells showed positive for CD2, CD5, CD7, CD34, CD56, HLA-DR, and cytoplasmic CD3. T cell receptor (TCR) and Immunoglobulin heavy chain genes were not rearranged. She received chemotherapy for acute lymphoblastic leukemia incorporating L-asparaginase (L-asp) which successfully induced complete remission. Bone marrow transplantation (BMT) from her HLA-identical sibling was conducted after two courses of consolidation therapy. Expression of aspargine synthetase (AS) protein in the leukemic cells at diagnosis was examined by an immunocytochemical method. She remains in hematological remission for over 36 months after BMT. The expression of AS protein was negative, suggesting that the leukemic cells were sensitive to L-asp. Induction and consolidation therapy incorporating L-asp followed by allo-BMT might be a promising treatment for child hood blastic NK cell leukemia, but more samples of the rare leukemia need to be studied before any definitive conclusions can be drawn. PMID- 15127421 TI - Cost-effectiveness acceptability curves--facts, fallacies and frequently asked questions. AB - Cost-effectiveness acceptability curves (CEACs) have been widely adopted as a method to quantify and graphically represent uncertainty in economic evaluation studies of health-care technologies. However, there remain some common fallacies regarding the nature and shape of CEACs that largely result from the 'textbook' illustration of the CEAC. This 'textbook' CEAC shows a smooth curve starting at probability 0, with an asymptote to 1 for higher money values of the health outcome (lambda). But this familiar 'ogive' shape which makes the 'textbook' CEAC look like a cumulative distribution function is just one special case of the CEAC. The reality is that the CEAC can take many shapes and turns because it is a graphic transformation from the cost-effectiveness plane, where the joint density of incremental costs and effects may 'straddle' quadrants with attendant discontinuities and asymptotes. In fact CEACs: (i) do not have to cut the y-axis at 0; (ii) do not have to asymptote to 1; (iii) are not always monotonically increasing in lambda; and (iv) do not represent cumulative distribution functions (cdfs). Within this paper we present a 'gallery' of CEACs in order to identify the fallacies and illustrate the facts surrounding the CEAC. The aim of the paper is to serve as a reference tool to accompany the increased use of CEACs within major medical journals. PMID- 15127422 TI - Should the consumption of survivors be included as a cost in cost-utility analysis? AB - Survivor costs are those costs associated with a treatment because it extends the patient's life. A controversy exists regarding whether survivor consumption costs should be included in cost-utility analyses. The present paper uses this controversy to motivate a general reexamination of what costs to include in cost utility analyses. Rather than the ad hoc inclusion rules currently used--a causal relationship between the intervention and the costs, and a proscription on double counting--this paper suggests three inclusion principles based on standard welfare economics. Thus, costs should be (1) included if they represent resources that directly produce the utility that is being measured in the denominator of the cost-utility ratio, (2) excluded if they represent resources that produce utility that is not being measured in the denominator, even though the costs are causally associated with the intervention, and (3) included if they represent resources consumed that are causally related to the intervention, but that have no counterveiling utility gains. These principles suggest important changes in how we account for recuperation time and unrelated medical care. They also suggest that survival consumption costs and earnings be excluded from existing cost-utility analyses. PMID- 15127423 TI - When does quality-adjusting life-years matter in cost-effectiveness analysis? AB - PURPOSE: This paper investigates the impact of quality-of-life adjustment on cost effectiveness analyses, by comparing ratios from published studies that have reported both incremental costs per (unadjusted) life-year and per quality adjusted life-year for the same intervention. METHODS: A systematic literature search identified 228 original cost-utility analyses published prior to 1998. Sixty-three of these analyses (173 ratio pairs) reported both cost/LY and cost/QALY ratios for the same intervention, from which we calculated medians and means, the difference between ratios (cost/LY minus cost/QALY) and between reciprocals of the ratios, and cost/LY as a percentage of the corresponding cost/QALY ratio. We also compared the ratios using rank-order correlation, and assessed the frequency with which quality-adjustment resulted in a ratio crossing the widely used cost-effectiveness thresholds of 20, 000 US dollars, 50,000 US dollars, and 100,000 US dollars/QALY or LY. RESULTS: The mean ratios were 69,100 US dollars/LY and 103,100 US dollars/QALY, with corresponding medians of 24,600 US dollars/LY and 20,400 US dollars/QALY. The mean difference between ratios was approximately -34,300 US dollars (median difference: 1300 US dollars), with 60% of ratio pairs differing by 10,000 US dollars/year or less. Mean difference between reciprocals was 59 (QA)LYs per million dollars (median: 2.1). The Spearman rank-order correlation between ratio types was 0.86 (p<0.001). Quality adjustment led to a ratio moving either above or below 50,000 US dollars/LY (or QALY) in 8% of ratio pairs, and across 100,000 US dollars in 6% of cases. CONCLUSIONS: In a sizable fraction of cost-utility analyses, quality adjusting did not substantially alter the estimated cost-effectiveness of an intervention, suggesting that sensitivity analyses using ad hoc adjustments or 'off-the-shelf' utility weights may be sufficient for many analyses. The collection of preference weight data should be subjected to the same scrutiny as other data inputs to cost effectiveness analyses, and should only be under-taken if the value of this information is likely to be greater than the cost of obtaining it. PMID- 15127424 TI - Does NICE have a cost-effectiveness threshold and what other factors influence its decisions? A binary choice analysis. AB - The decisions made by the National Institute for Clinical Excellence (NICE) give rise to two questions: how is cost-effectiveness evidence used to make judgements about the 'value for money' of health technologies? And how are factors other than cost-effectiveness taken into account? The aim of this paper is to explore NICE's cost-effectiveness threshold(s) and the tradeoffs between cost effectiveness and other factors apparent in its decisions. Binary choice analysis is used to reveal the preferences of NICE and to consider the consistency of its decisions. For each decision to accept or reject a technology, explanatory variables include: the cost per life year or per QALY gained; uncertainty regarding cost effectiveness; the net cost to the NHS; the burden of disease; the availability (or not) of alternative treatments; and specific factors indicated by NICE. Results support the broad notion of a threshold, where the probability of rejection increases as the cost per QALY increases. Cost effectiveness, together with uncertainty and the burden of disease, explain NICE decisions better than cost effectiveness alone. The results suggest a threshold somewhat higher than NICEs stated 'range of acceptable cost effectiveness' of pound 20,000 30,000 British pounds per QALY--although the exact meaning of a 'range' in this context remains unclear. PMID- 15127425 TI - Why cost-effectiveness should trump (clinical) effectiveness: the ethical economics of the South West quadrant. AB - In many health decision making situations there is a requirement that the effectiveness of interventions, usually their 'clinical' effectiveness, be established, as well as their cost-effectiveness. Often indeed this is effectively a prior requirement for their cost-effectiveness being investigated. If, however, one accepts the ethical argument for using a threshold incremental cost-effectiveness ratio (ICER) for interventions that are more effective but more costly (i.e. fall in the NE quadrant of the cost-effectiveness plane), one should apply the same decision rule in the SW quadrant, where the intervention is less effective but less costly. This implication is present in most standard treatments of cost-effectiveness analysis, including recent stochastic versions, and had gone relatively unquestioned within the discipline until the recent suggestion that the ICER threshold might be 'kinked'. A kinked threshold would, O'Brien et al. argue, better reflect the asymmetrical individual preferences found in empirical studies of consumer's willingness to pay and willingness to accept and justify different decision rules in the NE and SW quadrants. We reject the validity of such asymmetric preferences in the context of public health care decisions and consider and counter the two main 'ethical' objections that probably underlie the asymmetry in this case--the objection to 'taking away' and the objection to being required to undergo treatment that is less effective than no treatment at all. PMID- 15127426 TI - Regression methods for covariate adjustment and subgroup analysis for non censored cost-effectiveness data. AB - The current interest in undertaking cost-effectiveness analyses alongside clinical trials has lead to the increasing availability of patient-level data on both the costs and effectiveness of intervention. In a recent paper, we show how cost-effectiveness analysis can be undertaken in a regression framework. In the current paper we develop a direct regression approach to cost-effectiveness analysis by proposing the use of a system of seemingly unrelated regression equations to provide a more general method for prognostic factor adjustment with emphasis on sub-group analysis. This more general method can be used in either an incremental cost-effectiveness or an incremental net-benefit approach, and does not require that the set of independent variables for costs and effectiveness be the same. Furthermore, the method can exhibit efficiency gains over unrelated ordinary least squares regression. PMID- 15127427 TI - Stated and actual altruistic willingness to pay for insecticide-treated nets in Nigeria: validity of open-ended and binary with follow-up questions. AB - OBJECTIVES: To determine whether the binary with follow-up (BWFU) or open-ended (OPED) contingent valuation question format would yield better valid estimates of altruistic willingness to pay (WTP) and examine the feasibility of using intra community altruistic contributions to procure insecticide-treated nets (ITNs) for the poor in Nigeria. METHODS: Structured questionnaires were used to elicit stated altruistic WTP from a random sub-sample of respondents that had either the OPED or BWFU used to elicit WTP in Southeast Nigeria. One month after the survey the respondents were asked to redeem their WTP pledges. Construct validity was determined using econometric analyses, while phi correlation coefficient was used to determine criterion validity. FINDINGS: More than 57% of the respondents were hypothetically willing to pay for altruism in both BWFU and OPED groups. Altruistic WTP was positively related to respondents' WTP for own nets (p<0.01) and nets for other household members (p<0.05) in both groups. A total of 27.0% and 33.1% of the respondents with positive hypothetical WTP in the BWFU and OPED actually contributed. Phi correlation coefficient was 0.23 (95%CI 0.20-0.29) in BWFU and 0.49 (95%CI 0.44-0.54) in OPED. The money realised was used to buy some ITNs, which were presented to poor people selected by community leaders. CONCLUSION: The OPED elicited better valid estimates of altruistic WTP than BWFU. The potential for more capable people to contribute for the poor in ITNs programmes actually exist and malaria control programmes should explore altruistic contributions as a means to increase net coverage. PMID- 15127428 TI - Comorbidities and the willingness to pay for reducing the risk of a targeted disease: introducing endogenous effort for risk reduction. AB - Previous studies demonstrated that the willingness to pay (WTP) for reducing the risk of a targeted disease increases with the severity of a comorbidity condition. They evaluated the WTPs at positions where otherwise identical individuals who have unequal comorbidity conditions all have the same consumption of wealth and the same probability of getting a targeted disease. This paper endogenizes the individual effort to reduce the risk of the targeted disease. It finds that individuals with more severe comorbidity will spend more on reducing the risk of the targeted disease and, therefore, have a lower probability of getting the targeted disease and a lower net consumption of wealth at equilibrium. Nonetheless, we find that the previous conclusion concerning the relationship between the WTP for the targeted risk reduction and the comorbidity is robust. PMID- 15127431 TI - Polyamides as artificial transcription factors: novel tools for molecular medicine? PMID- 15127432 TI - Electrochemistry/mass spectrometry (EC/MS)--a new tool to study drug metabolism and reaction mechanisms. PMID- 15127433 TI - Nanolithography and nanochemistry: probe-related patterning techniques and chemical modification for nanometer-sized devices. AB - The size regime for devices produced by photolithographic techniques is limited. Therefore, other patterning techniques have been intensively studied to create smaller structures. Scanning-probe-based patterning techniques, such as dip-pen lithography, local force-induced patterning, and local-probe oxidation-based techniques are highly promising because of their relative ease and widespread availability. The latter of these is especially interesting because of the possibility of producing nanopatterns for a broad range of chemical and physical modification and functionalization processes; both the production of nanometer sized electronic devices and the formation of devices involving (bio)molecular recognition and sensor applications is possible. This Review highlights the development of various scanning probe systems and the possibilities of local oxidation methods, as well as giving an overview of state-of-the-art nanometer sized devices, and a view of future development. PMID- 15127434 TI - Screening of chiral catalysts and catalyst mixtures by mass spectrometric monitoring of catalytic intermediates. PMID- 15127435 TI - Remarkable 4,4'-substituent effects on binap: Highly enantioselective Ru catalysts for asymmetric hydrogenation of beta-aryl ketoesters and their immobilization in room-temperature ionic liquids. PMID- 15127436 TI - Semisynthesis of a functional K+ channel. PMID- 15127438 TI - Visual observations of SERRS from single silver-coated silica microparticles within optical tweezers. PMID- 15127437 TI - A droplet-based, composite PDMS/glass capillary microfluidic system for evaluating protein crystallization conditions by microbatch and vapor-diffusion methods with on-chip X-ray diffraction. PMID- 15127439 TI - Probing the mechanism of the Heck reaction with arene diazonium salts by electrospray mass and tandem mass spectrometry. PMID- 15127440 TI - Glycosidation reactions of silyl ethers with conformationally inverted donors derived from glucuronic acid: stereoselective synthesis of glycosides and 2 deoxyglycosides. PMID- 15127441 TI - A monomeric organolithium compound containing a free pyramidal carbanion in solution and in the solid state. PMID- 15127442 TI - A mild method for the preparation of gamma-hydroxy-alpha,beta-acetylenic esters. PMID- 15127443 TI - Size-selective olefin hydrogenation by a Pd nanocluster provided in an apo ferritin cage. PMID- 15127444 TI - Reaction-temperature-dependent supramolecular isomerism of coordination networks based on the organometallic building block [CuI2(mu2-BQ)(mu2-OAc)2]. PMID- 15127445 TI - A one-pot total synthesis of crambin. PMID- 15127446 TI - Nanocrystalline CeO2 increases the activity of Au for CO oxidation by two orders of magnitude. PMID- 15127447 TI - Sb8(GaCl4)2: Isolation of a homopolyatomic antimony cation. PMID- 15127448 TI - Highly enantioselective construction of fused pyrrolidine systems that contain a quaternary stereocenter: concise formal synthesis of (+)-conessine. PMID- 15127449 TI - The cofactor of the iron-sulfur cluster free hydrogenase hmd: structure of the light-inactivation product. PMID- 15127451 TI - Significant interactions between uranium and noble-gas atoms: coordination of the UO2+ cation by Ne, Ar, Kr, and Xe atoms. PMID- 15127450 TI - Isolation and characterization of pre-rapamycin, the first macrocyclic intermediate in the biosynthesis of the immunosuppressant rapamycin by S. hygroscopicus. PMID- 15127453 TI - In pursuit of carbohydrate-based HIV vaccines, part 2: The total synthesis of high-mannose-type gp120 fragments--evaluation of strategies directed to maximal convergence. PMID- 15127452 TI - In pursuit of carbohydrate-based HIV vaccines, part 1: The total synthesis of hybrid-type gp120 fragments. PMID- 15127454 TI - Stereochemical control of the redox potential of tetracoordinate rhodium complexes. PMID- 15127455 TI - Highly resolved spin-density distribution in the Prussian-blue precursors Cs2K[Fe(CN)6] and Cs2K[Mn(CN)6]. PMID- 15127456 TI - Abyssomicin C-A polycyclic antibiotic from a marine Verrucosispora strain as an inhibitor of the p-aminobenzoic acid/tetrahydrofolate biosynthesis pathway. PMID- 15127458 TI - Breast cancer survivors: learning from the faces of hope. PMID- 15127459 TI - Sharing the secret: talking about urinary incontinence. PMID- 15127461 TI - Tough times call for caring leaders. PMID- 15127462 TI - Education expectations more consistent. AB - The Joint Commission has revised core standards in the Education chapter of the Comprehensive Accreditation Manual for Hospitals. PMID- 15127463 TI - Who wants to spend $1,000? AB - When it comes to capital purchases, your opinion does count. PMID- 15127464 TI - Stop the rise in nursing errors--systematically. AB - The number of reported nursing errors in hospitals has increased in each of the past 5 years. Information technology can help nurses make better care decisions in today's harried hospital environment. PMID- 15127465 TI - Long-term care demands precise documentation. AB - It's more common for the elderly to die in long-term care versus acute care. Their needs are also far more varied, as is the facility's liability exposure. Learn how caregivers' documentation can save lives and reduce liability. PMID- 15127466 TI - I.v. infiltration. Not just a peripheral problem. AB - Learn how to recognize and treat infiltration and extravasation at various peripheral and central intravenous sites. PMID- 15127467 TI - Role call: 6 months to better nurses. PMID- 15127468 TI - Your hand in glove selection. AB - Supplies that are purchased for facility-wide use can have a sizeable impact on a hospital's bottom line as well as patient and employee safety. Here, review how to maximize your purchasing power when selecting medical gloves. PMID- 15127469 TI - ECT in the PACU? It's possible. AB - Follow how this facility established and supported an electroconvulsive therapy program in its postanesthesia care unit, including multidisciplinary communication and patient and staff education. PMID- 15127470 TI - High-tech learning means more access, more participation--and more nurses. AB - Technology has revolutionized nursing education. Learn how distance learning, high-tech teaching aids, and self-directed courses can benefit you and your staff. PMID- 15127471 TI - Trail the state board's complaints. AB - If your state board of nursing receives a complaint about one of your nurses, the complaint triggers a structured disciplinary process. Follow the process from start to finish and test your knowledge, then check your answers at http://www.nursingmanagement.com. PMID- 15127472 TI - Nurses--yes, nurses--improve physician order entry. AB - Automated physician order entry systems can reduce medication errors and improve patient outcomes. But there are a variety of barriers--psychological, behavioral and technological--to physician adoption. Nursing not only has a stake in this issue, but plays a valuable role in resolving it. PMID- 15127473 TI - Surveys go full-service. AB - In response to health care organizations' increasing diversity of services, the Joint Commission has developed an integrated survey process. PMID- 15127474 TI - A lifestyle how-to for night-shift nurses. AB - Lifestyle training teaches night-shift nurses the factors that impact their alertness and helps them adjust their lives to improve it. Learn how to structure a program for your nurses. PMID- 15127475 TI - A wake-up call. PMID- 15127478 TI - Tackling the issue of nurse competency. AB - Learn how to ensure and enhance your staff's competency using the TEAM concept. PMID- 15127479 TI - Staying on top of TB's rise. AB - Delays in acid-fast bacillis (AFB) emergency department admissions prompted a revised multidisciplinary rounds process for weekends and holidays; AFB isolation cases dropped 10%. PMID- 15127480 TI - AFB isolation rounds: what your nurses need to know. PMID- 15127481 TI - Dignity in dying. AB - Barriers to effective end-of-life care encompass patient, professional, and system issues. Learn the findings of a landmark study on how well nurses meet the needs of dying patients. PMID- 15127482 TI - Taking the path less traveled. AB - Behavioral health is a new area for clinical pathways. Learn the steps to develop one, the pitfalls and benefits, and how to support your continuous quality improvement efforts. PMID- 15127483 TI - Stopping the word war. AB - Find out what's behind interdisciplinary anger and how to teach nurses to break this vicious cycle. PMID- 15127484 TI - Addressing wounds. AB - Learn your options for dressing wounds from the following overview. PMID- 15127485 TI - Defeating malpractice risk, Part 2. AB - Part 1 explored the basic elements of malpractice--duty, breach of duty, injury, and causation--and how to reduce malpractice risk for medication and equipment errors. This month, learn the malpractice risks and cautions for monitoring mistakes and patient falls. Test your knowledge with the questions below, then check your answers at http://www.nursingmanagement.com. PMID- 15127486 TI - Putting you to the test. AB - New to nursing leadership? Know someone who is? This new bimonthly department's advice on how to manage employees and set practice expectations can help. PMID- 15127487 TI - New rules impact sedation and anesthesia care, Part 1. AB - Learn why the Joint Commission has revised its anesthesia care standards. The new standards incorporate four sedation levels: minimum sedation, moderate sedation/analgesia, deep sedation/analgesia, and anesthesia. PMID- 15127488 TI - 10 real-time alerts! AB - A computerized patient record system that provides real-time clinical decision support lets you set rules for clinical alerts. Learn the 10 ways you can make the system work for you. PMID- 15127489 TI - Recruitment and retention report. Hidden charges. PMID- 15127490 TI - Wise buys. Do you have what it takes to sway new product purchases? AB - Learn the three competencies you need to make the best purchasing decisions: end user accountability, nursing commercial competence, and evidence-based product selection. PMID- 15127491 TI - Telltale signs. AB - Educate your staff to identify and document the behavioral, physical, and performance-related signs of drug and alcohol dependency. Then help your dependent employee and her peers deal with the problem and move on. PMID- 15127492 TI - If the gut works use it. AB - Using education and increased monitoring, a nutritional support team lowered parenteral nutrition days from 84 to 5 per quarter, increased the proportion of patients meeting their nutritional requirements from 75% to 97%, and saved money. PMID- 15127493 TI - Survivor's syndrome. AB - Poor morals due to layoffs is the health care industry's human resources problem. Learn the psychological responses of survivor's syndrome, how to identify it in your employees, and now to turn their energy toward the newly restructured organization. PMID- 15127494 TI - Families first. AB - Family support can have positive effects on critically ill patients, so limiting visitation in critical care units may not be in their best interest. This study examined the outcomes of increasing visitation using a quality-improvement model. PMID- 15127495 TI - Vital signs' vital integration. AB - Vital signs monitoring systems need to pool information from a variety of bedside devices. Add options like portability and multiple parameters, and you can choose the best one for any setting. PMID- 15127497 TI - The three C's: cloudy or clear? AB - Learn what standards the Joint Commission sets for communication, coordination, and continuum of care. PMID- 15127498 TI - Should your data collection expand or shrink? AB - To get the most out of data collection, here's how to analyze its purpose, application, and cost-effectiveness. PMID- 15127499 TI - Don't go! AB - Director of nursing (DON) turnover can be high, particularly in rural areas. To address it at your facility, learn the conditions that influence DON turnover. PMID- 15127500 TI - Providing a safe environment in the new millennium. PMID- 15127501 TI - Staffing: know your liability. AB - Read this case of poor staffing judgment regarding outside agency employees and answer the questions, then check your answers at http://www.nursingmanagement.com. PMID- 15127502 TI - Exploring the gray areas of informed consent. AB - Learn your role in informed consent and explore the gray areas, including circumstances when consent may be valid even if the patient wasn't fully informed. PMID- 15127503 TI - Out in front: a new title reflects nurse managers' changing scope of accountabilities. AB - A new leadership team surveys health care practitioners to assess and redesign the front-line manager role in its merged system. PMID- 15127504 TI - Salary survey 2000. PMID- 15127505 TI - Clinical information systems. Do-it-yourself guide. AB - In a 200-bed community hospital, nurse leaders led a team to design, program, and implement a new clinical information system. PMID- 15127506 TI - Put power in nurses' hands. AB - Orlando's Nursing Theory gives nurses an interpersonal, straightforward approach to either simple or complex situations. They can apply the theory individually or as a department--even with time, energy, and financial constraints. PMID- 15127507 TI - Help from our animal friends. AB - Animal-assisted therapy has gained widespread support and application in the past few decades. Learn its present goals and applications, including those pertinent to critical care. PMID- 15127508 TI - Clearing the way for a CPR. AB - While waiting for a universal computer-based patient record, many hospitals have already implemented one through their clinical information systems. Here's a review of the potential benefits. PMID- 15127509 TI - What are your rules of engagement? PMID- 15127510 TI - XML: health care's killer application? AB - XML promises to be the next big thing in health care information. PMID- 15127511 TI - We lead in interesting times. PMID- 15127512 TI - Master five common medication administration questions. AB - A look at some of the most frequently asked questions regarding the Joint Commission's standards on medication administration. PMID- 15127513 TI - Shaping the medical error movement. AB - The 1999 Institute of Medicine's report. "To Err is Human," shifted government and public focus to medical errors. Read how errors occur and what's being done to stop them, then check your answers at http://www.nursingmanagement.com. PMID- 15127514 TI - Pass it on? AB - Pointers for managers to ensure safe, successful delegating practices. PMID- 15127515 TI - 2 cents for products under $100. AB - For many of the more than 25,000 hospital products under $100, nurses are the end users--and the best source of evaluation. Teach nurses how to make their voices heard. PMID- 15127517 TI - No nurse is an island. PMID- 15127516 TI - Staying in the lines. AB - Nurses must set professional boundaries for their relationships with patients. You can teach them the meaning of boundary crossings, boundary violations, and staying in the lines. PMID- 15127518 TI - Boomers vs. busters: bridging the generation gap. PMID- 15127519 TI - Start a freestanding emergency center. AB - Community need sparked the concept for a freestanding emergency center. Key first steps: new legislation, construction, and education. PMID- 15127520 TI - Cost-justifying ED technology. AB - Weigh your options carefully when purchasing new products for the ED. Here are some pointers to ensure a smart buy. PMID- 15127521 TI - How do we report drug abuse? PMID- 15127522 TI - Mission with a margin. PMID- 15127523 TI - Making a nurse intern program pay off. AB - A nurse intern program is expensive but has several big payoffs: recruitment, retention, and well-trained, satisfied nurses. PMID- 15127524 TI - Medical errors, airplanes, and information technology. AB - The IOM says tens of thousands die each year from medical errors. Learn lessons from the aviation industry and how we can build a culture of safety with information technology and clinical information systems. PMID- 15127525 TI - New rules impact sedation and anesthesia care, Part 2. AB - In Part 1, the author explained the four new definitions that are changing the Joint Commission's sedation and anesthesia standards: minimum sedation, moderate sedation/analgesia, deep sedation/analgesia, and anesthesia. Part 2 explains how these definitions will impact the delivery of care in your units. PMID- 15127526 TI - Negligence goes to the top. AB - Nurse managers aren't only responsible for their own actions and those of their nurses but also for recognizing and acting on physicians' inappropriate care. Learn from a case where nurses failed to take action throughout a patient's care- and the cost was her life. PMID- 15127527 TI - "Hospitable" hospitals. PMID- 15127528 TI - Legislating sharps safety. AB - Various state laws, passed and pending, stress the need for federal legislation on needle-stick prevention. PMID- 15127529 TI - Minimizing the threat of C. difficile. AB - Learn the signs and symptoms of Clostridium difficile-associated diarrhea (CDAD), outline infection control measures to help stop its spread and review treatment regimens. PMID- 15127530 TI - Strike a balance with decentralized housekeeping. AB - Housekeeping staff join patient-care teams to improve patient care, but the transition can cause problems of its own. Read how one hospital minimized the negative effects and improved staff productivity and patient satisfaction. PMID- 15127531 TI - Restraints rock. AB - Restraint issues can create problems for staff, patients, and family. To meet the Joint Commission's restraint rules, a multidisciplinary education plan included alternative and distractive devices for patients. Use of restraints decreased markedly as staff members' knowledge and attitude improved. PMID- 15127532 TI - Good use of short time. AB - Although outpatient surgery shortens hospital stays, patients still experience anxiety and pain. Encourage nurses to maximize their communication skills and focus on pain management. PMID- 15127533 TI - New catheters decrease nosocomial infections. AB - Urinary tract infections and intravascular infections cause pain or even death, delay healing, and raise costs. New technology makes catheters safer than ever, so you can reduce these infections in your departments. PMID- 15127534 TI - The nurse leader's credo: think globally, act locally. PMID- 15127535 TI - New standards support nursing's licensed independent practitioners. AB - New standards give nurses who work as licensed independent practitioners recourse when an organization limits or revokes privileges. PMID- 15127536 TI - Beyond hospital walls. AB - The Health Care Financing Administration plans to expand hospital response boundaries to a 250-yard radius of the facility. Learn how this will affect you, then check your answers at http://www.nursingmanagement.com. PMID- 15127537 TI - Preventing postoperative complications. AB - Learn nursing interventions for preventing or dealing with some common postoperative complications--including how the latest research may affect standard techniques. PMID- 15127538 TI - Upbeat advances in cardiac and emergency care. AB - From biphasic breakthroughs to simplified programming mechanisms, review the latest technology options. PMID- 15127539 TI - In our hands? The future of health care technology. AB - Use improvements in data accuracy and nursing productivity to enhance your quality of care. PMID- 15127540 TI - Get the right information to the right people at the right time. AB - With so much information and so many systems available to handle it, finding the right solutions for your units can get complicated. Start with the basics of nurse communication, then find your best fit. PMID- 15127541 TI - Where it's at: point-of-care testing yields results--stat. AB - Discover bedside technology breakthroughs that increase health care safety, accuracy, speed, and convenience. PMID- 15127542 TI - Breakthroughs in vascular access. AB - Learn how new vascular products prevent needlesticks, infection, and vein thrombosis. PMID- 15127543 TI - Get a handle on safe patient transfer and activity. AB - Transferring patients is a high-risk activity. Learn how the proper equipment can ease this process. PMID- 15127544 TI - ICP technology keeps germs in check. AB - Learn how the convenience of waterless cleansers and the Internet can assist you and your staff with germ control. PMID- 15127545 TI - Nursing and respiratory: partners in care. AB - Learn about RCP licensure and nursing's support of this process, as well as the clinical advancements, new and altered procedures, and novel applications of drugs and devices in respiratory care. PMID- 15127546 TI - Be a watchdog. AB - Review the advances in computerized prescriber order entry and automated storage and administration devices, which effectively reduce medication errors. PMID- 15127547 TI - Oh no! The boss has gone "psycho!". PMID- 15127548 TI - Prepare for your best employee evaluation yet. AB - Don't get nervous, get ready! Prepare your employee, your setting, and yourself to get the most out of employee evaluations. PMID- 15127549 TI - The ABCs of APCs. AB - To make sure your ED has money to spend, you need an Ambulatory Payment Classifications process that's streamlined, error-free, and compliant with regulations. Here's how. PMID- 15127550 TI - Revisions cut restraint risk. AB - The Joint Commission has revised its restraint and seclusion standards for behavioral health facilities and hospitals, effective January 1, 2001. The standards protect patients and staff, stress the importance of training, and clarify caregivers' roles. PMID- 15127551 TI - Don't get underrepresented. AB - Beware of the situations that can arise when a nurse fails to retain an attorney, hires a generalist when the case requires a specialist, or shares an attorney with other parties in the same case. Check your answers to the following questions at http://www.nursingmanagement.com. PMID- 15127552 TI - Nurses get IT right. AB - When nurses get involved in IT selection, the resulting system supports their work and saves the organization time and money. PMID- 15127553 TI - Buzzwords with a basis. AB - Learn the essential elements that characterize the concepts of motivation, mentoring, and empowerment, and explore the integral relationships between and among the three. PMID- 15127554 TI - From positive to parenthood. AB - Perinatal care coordinators track patients throughout pregnancy and provide a seamless admission to the labor and delivery unit. Patients benefit from early referral to community resources and education about birth options. PMID- 15127555 TI - The making of a supermodel. AB - This model serves as a mechanism for improved care delivery and ensures consistent patient care experience at each affiliate. PMID- 15127556 TI - Managing i.v. therapy: "high-alert" drugs keep nurse managers ever watchful. AB - Learn techniques critical for safe i.v. infusion and the appropriate steps to take before, during, and after administration of five "high-alert" medications- potassium chloride, heparin, dopamine, theophylline, and insulin. PMID- 15127557 TI - An outcomes approach to skill mix change in critical care. AB - When the nursing shortage strained the resources of one community hospital, RNs partnered with LVNs to drop costs, raise patient satisfaction levels, and maintain quality patient care. PMID- 15127558 TI - A cc of prevention. AB - Rarely do saving money and saving lives fit so closely together as they do with sharps safety. Here's how to choose the best devices to reduce injuries in your hospital. PMID- 15127559 TI - [Relationship between susceptibility and prognosis of laryngeal cancer and genetic polymorphisms in CYP1A1 and GSTM1]. AB - OBJECTIVE: To investigate the relationship between susceptibility and prognosis of laryngeal cancer and genetic polymorphisms in CYP1A1 GSTM1. METHODS: The genetic polymorphisms of CYP1A1 exon-7 and GSTM1 were analyzed by polymorphism polymerase chain reaction (PCR) technique in peripheral blood leukocytes of 89 laryngeal cancer cases and 164 non-cancer controls. RESULTS: Either in the frequency of CYP1A1 (W, MW, M) and GSTM1 negative individual of laryngeal cancer were significantly higher than that in the controls(P < 0.05). Combined genetype of CYP1A1 and GSTM1, that is CYP1A1 W/GSTM1 (-), CYP1A1 MW/GSTM1 (-), CYP1A1 M/GSTM1 (+), CYP1A1 M/GSTM1 (-) had higher risk than those combined genotypes, which ratios were 1.42 (0.62-3.22), 1.67 (0.7-4.03), 2.55 (1.0-6.54) and 4.02 (1.65-9.79), respectively. Statistic analysis suggested an interaction between smoking and CYP1A1 M/GSTM1 (-) genotypes polymorphisms which increase risk of laryngeal cancer. There was no significant difference in the frequency of CYP1A1, GSTM1 individual with the development and prognosis of laryngeal cancer. CONCLUSION: The present datas suggested that genetic polymorphisms of CYP1A1 and GSTM1 were susceptible to laryngeal cancer. Individuals with CYP1A1 M or GSTM1 ( ) had increased risk of developing laryngeal cancer, but individuals with combined CYP1A1 M/GSTM1 (-) and smoking were more susceptible. Genetic polymorphisms of CYP1A1 and GSTM1 are not related with clinic process of laryngeal cancer. PMID- 15127560 TI - [Human antisense-vascular endothelial growth factor gene therapy for laryngeal tumor by cationic liposome-mediated transfection]. AB - OBJECTIVE: To observe the effect of cationic liposome mediated antisense-vascular endothelial growth factor (VEGF) gene transfection on the growth of laryngeal cancer Hep-2 cells in the nude mice. METHODS: The VEGF-cDNA gene was cloned by reverse transcriptase polymerase chain reaction (RT-PCR) from human laryngeal cancer, and its eukaryotic expression vector pcDNA3-VEGF (-) with antisense-VEGF gene was constructed and identified by PCR and double-enzyme digestion. The pcDNA3-VEGF (-) was transfected into laryngeal cancer Hep-2 cell line by using cationic liposome (LP 2000). Then, the transfected Hep-2 cells were injected into nude mice and the size of tumor from different groups was observed while establishing laryngeal cancer xenografts in nude mice, and then treating the tumor-bearing mice with liposome-plasmid complex, observing the size of tumor from different groups. The expression of VEGF mRNA in different groups was observed by RT-PCR. The transfected cell ultranstructure was observed by transmission electron microscopy. RESULTS: The human VEGF-cDNA was successfully cloned and its eukaryotic expression vector with antisense-VEGF pcDNA3-VEGF (-) was constructed. The antisense-VEGF gene was transfected into Hep-2 cell line by using cationic liposome (LP2000). The size of tumor transfected with pcDNA3-VEGF (-) was significantly smaller than that of control groups. While the size of tumor treated with liposome-pcDNA3-VEGF (-) complex was significantly smaller than that of control groups. Many apoptic tumor cells were observed by transmission electron microscopy and the structure of microvessel was also changed. The expression of VEGF mRNA was evidently weaker than that of the control groups. CONCLUSION: The growth of Hep-2 cells could be inhibited significantly by antisense-VEGF gene transfection. PMID- 15127561 TI - [Detection of mutation in exon 5 and exon 8 of PTEN in laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To detect mutations of exon 5 and exon 8 of phosphatase and tensin homology deleted on chromosome10/mutated in multiple advanced cancer1/TGF-beta regulated and epithelial cell-enriched phosphatase (PTEN/MMAC1/TEP1 for short PTEN) gene in laryngeal squamous cell carcinoma (LSCC) and analyze the relationship between the mutation and LSCC. METHODS: Fresh tumor samples from 40 LSCC patients were examined using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and direct DNA sequencing. RESULTS: The mutation of exon 5 and exon 8 occurred in supra-glottic laryngeal carcinoma and no mutation in the glottic laryngeal carcinoma was found. There were 6 cases of mutation for exon 5, and the mutation rate was 15%. In 2 cases, 1 base pair insertion TT-->TAT in codon 85, while in other two cases, 1 base pair deletion GCA-->GC in codon 86, and the two type mutation may result in frame shift mutation. One case had the above two type mutation simultaneously, which may result in missense mutation; 1 case had 1 base pair insertion TT-->TAT in 85 codon and 1 base pair deletion GCA-->GA, which may result in nonsense mutation. The pathology of 3/6 (50%) cases was low differentiation, and the others (3/6, 50%) were middle differentiation. Five cases had lymphatic metastasis, and the rate of which was 83.3%; the other one had no lymphatic metastasis, and the rate of which was 16.7%. There was only 1 case mutation for exon 8, and the mutation rate was 2.5%. One base pair deletion AAA-->AA in 276 codon as well as 1 base pair insertion CT-->CCT between the 336 codon and 337 codon may result in frame shift mutation. The pathology of the case was low differentiation. CONCLUSION: The codon 85,86 of PTEN gene in exon 5 may be "hot spots" in LSCC and the mutation of PTEN gene may be related with the lymphatic metastasis and middle or low differentiation. PMID- 15127562 TI - [Survival analysis of 1115 patients with laryngeal carcinoma]. AB - OBJECTIVE: To study the long-term result of patients with laryngeal carcinoma treated by surgery and the prognostic factors. METHODS: The survival status of 1115 patients with laryngeal carcinoma which were treated in this department between 1983 and 1996 were retrospectively investigated. RESULTS: Overall 5-year survival rate was 77%. Among them, 5-year survival rate for patients of stage I was 94%, stage II 89%, stage III 82%, stage IV 66%. Patients with glottic cancer had the best prognosis, followed by supraglottic, subglottic and transglottic cancer. Five years survival rate for patients with partial laryngectomy was 85%, whereas, for total laryngectomy was 68%. The causes of failure were local recurrence and metastasis(70%). 14% of dead was not clear. CONCLUSION: Early diagnosis was the key for both larynx preservation and survival rates. Causes of dead were laryngeal recurrence and metastasis. PMID- 15127563 TI - [Combined treatment for advanced laryngeal carcinoma]. AB - OBJECTIVE: To assess the efficacy of the combined therapy for advanced laryngeal carcinoma. METHODS: 204 cases of advanced laryngeal carcinoma were treated in this department from 1984 to 1997. Patients were treated by surgery alone or combined therapy including of surgery, radiotherapy and chemotherapy. RESULTS: The overall 3 and 5 year survival were 70.1% (143/204) and 61.8% (126/204) respectively. 5-year survival rate of the combined therapy group was 68.2% (90/132), whereas, 5-year survival was only 50.0% (36/72) for surgery alone group (P < 0.05). There was no statistical differences between surgery alone group and the combined therapy group in the local recurrences (P > 0.05) but distant metastases (P < 0.05). There were also no statistical differences between the preoperative radiotherapy group and the non-preoperative radiotherapy group in the terms of infection and fistula (P > 0.05). CONCLUSION: The results suggest that the combined therapy was more effective than surgery alone for the treatment of the advanced laryngeal carcinoma. Preoperative radiotherapy did not increase the incidences of infection and fistula. PMID- 15127564 TI - [Cervical metastasis in patients with T2-4 cN0 laryngeal carcinoma]. AB - OBJECTIVE: To study the characteristics of the cervical lymph node metastasis in clinical N0 (cN0) patients with laryngeal carcinoma and its implication in clinical treatment. METHODS: 76 patients with laryngeal carcinomas of T2-4cN0 category were divided into two groups in random: 21(22 sides) radical neck dissection(RND) and 55(60 sides) functional neck dissection(FND) were performed. Lymph nodes were studied histologically according to the levels. RESULTS: On an average, 29.6 lymph nodes were obtained in one side of neck in RND group, and 24.7 in FND group(F = 3.145, P = 0.068). The occult metastasis rates were 33.3% (7/21) in RND group and 34.5% (19/55) in FND group. 25 of 26 patients (96.2%) who had positive nodes involved only the levels II and III. 2130 lymph nodes were obtained in all samples, 59 of 60 positive nodes(98.3%) were located in the level II and III. The 5 and 10-year survival rates of the two groups were 71.4% (15/21), 76.4% (42/55) and 61.9% (13/21), 68.9% (31/45), respectively with no statistical difference(chi 2 = 0.2394, P > 0.5; chi 2 = 0.3143, P > 0.05). Ipsilateral cervical recurrence rates in two groups were 9.5% (2/21) and 7.3% (4/55), respectively with no statistical difference (chi 2 = 0.1059, P > 0.900). 10-year mortalities with negative and positive cervical lymph nodes were 16.7% (7/42) and 62.5% (15/24) respectively, which had statistically difference (chi 2 = 14.4375, P < 0.005). CONCLUSION: The lateral neck (level II, III and IV) dissection may be suitable for the treatment laryngeal carcinoma patients with T2 4cN0. PMID- 15127565 TI - [Functional evaluation of the selective neck dissection in patients with carcinoma of head and neck]. AB - OBJECTIVE: To investigate comparatively the shoulder function, sensory and cosmetic changes after selective neck dissection (SND), modified neck dissection (MND) or radical neck dissection (RND). METHODS: 157 questionnaires, which involving shoulder function, skin numbness and cosmetic changes were sent to patients with SCC of head and neck who were treated with SND, MND or RND between January 1997 and May 2001. 32 eligible questionnaires were collected which included 32 patients had 43 neck dissection. Among them, SNDs were performed in 23 necks, MNDs in 11 necks and RNDs in 9 necks. RESULTS: The shoulder disability was least in SND necks (13.0%), followed by MND necks (36.6%) and RND necks (66.7%). Similarly, permanent neck anesthetic were found in 17.4% of SND necks, 36.6% of MND necks, and 88.9% of RND necks. Last, only 17.4% of SND necks had cosmetic changes after surgery, while 90.9% of MND necks and 100% of RND necks had cosmetic changes after surgery. CONCLUSION: Patients underwent selective neck dissection had the least damage to the shoulder function, skin sensory and cosmetic. PMID- 15127567 TI - [Electrically stimulated olfactory evoked potential in olfactory-lesioned rabbit]. AB - OBJECTIVE: To develop olfactory dysfunctioned animal model by injuring olfactory blub(s) and to investigate the characteristics of olfactory evoked potential. METHODS: The changes of OEP in rabbits were observed at 24 h, 48 h and 1 w after olfactory bulb(s) were electrolytically injured. The pathological and ultrastructural changes of olfactory mucosa and olfactory bulb were also observed. RESULTS: When unilateral olfactory bulb was damaged, N2 vanished with elongated latencies and lowered amplitudes of N1 and P1. When both olfactory bulbs were damaged, N1 and N2 were absent, only P1 could be recorded. The latency was extended obviously and the amplitude had a big change. Histopathology and ultrastructure showed that there were a lot of inflammatory cells and degenerated neurons scattered around OB 24 hours to 48 hours after injure. It was observed that the olfactory cilia had distraction perversion and colliquefaction changes. CONCLUSION: Different degree of olfactory bulb injury caused different effect on OEPs. These changes of potential were based on the histopathology and ultrastructure. PMID- 15127566 TI - [Quantification of endolymphatic hydrops in experimental animal model]. AB - OBJECTIVE: To investigate a method of quantification of the endolymphatic hydrops in experimental animal model. METHODS: Thirty guinea pigs were divided into three groups at random. In control group, there were ten guinea pigs without operation on both ears. Endolymphatic hydrops was induced by endolymphatic sac obliteration through extradural posterior cranial fossa approach in right ear, including 4 week postoperative group(n = 10) and 8-week postoperative group(n = 10). The area of scala vestibuli (SV) and scala media(SM) of each turn on both cochlear midmodiolar sections was measured, respectively, using auto computer aided design(AutoCAD R14) software combined with digital camera, and then the maximum scala media area(SMA) ratio was calculated and compared. RESULTS: No endolymphatic hydrops was observed in all non-operated ears, however, variety degree of hydrops was present in all operated ears. The average maximum SMA ratio in the 4-week group (2.2231 +/- 0.1996) was greater than that in the control group (1.0971 +/- 0.0644). The average maximum SMA ratio of the 8-week group (4.0142 +/- 0.5218) was greater than that in the 4-weekgroup. There was significant difference between the two groups(P < 0.05). CONCLUSION: It is convenience and reliable to be used to quantify the experimental endolymphatic hydrops with the present method. This study provides a reliable methodological base for the experimental study of Meniere's disease. PMID- 15127568 TI - [Operative therapy of the adhesive otitis media]. AB - OBJECTIVE: To study the operative indications and treatment for adhesive otitis media (AOM). METHODS: Sixty-nine cases of AOM were undergone tympanoplasty between 1995 to 2000. Preoperative average air-bone gap(A-B gap) were (43.7 +/- 3.9) dB. Forty-one cases could be postoperative followed up for more than one year. Preoperative average A-B gap of this group were (43.8 +/- 4.2) dB. The adhesive ear drum and the other processes in middle ear were inserted and reconstructed with cartilage. RESULTS: The postoperative average A-B gap after one month(69 cases) and one year(41 cases) were (23.7 +/- 8.6) dB and (27.9 +/- 10.7) dB. In the long-time follow-up group, the A-B gap of 18 cases (43.9%) were less than 20 dB. Twenty-four cases (58.5%) become a nearly normal tympanum. CONCLUSION: The AOM can treatment with tympanoplasty. But the indication must be carefully selected. The cartilage was a good material for reconstruction of the ear drum to treatment the AOM. The influence factor were analyzed. PMID- 15127569 TI - [Videomimicography: a new objective evaluation of facial motor function]. AB - OBJECTIVE: To develop a new method, videomimicography (VMG), to evaluate facial motor function. METHODS: During VMG, 11 landmarks were placed on the face, and 5 movements (forehead lifting, eye closure, nose wrinkling, lip puckering and smiling) with maximal contraction were requested. A digital video film was recorded then fed in a computer to be analyzed. Ten normal subjects were used as normal control. RESULTS: Area measures were found better than distance measures in evaluating facial movements. The best measure for each movement was determined. Then a global index of facial motor function was derived from these measures. This index was found well correlated with the facial paralysis House Brackmann grade in 48 patients with facial paralysis (r = -0.928). CONCLUSION: VMG is an objective, quantitative, relative simple method. It has good reproducibility. So it can be used in clinic for evaluating facial motor function. PMID- 15127570 TI - [Peri-uvulopalatopharyng oplasty otoacoustic emissions in patients with obstructive sleep apnea-hypopnea syndrome]. AB - OBJECTIVE: To study the cochlear function of patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). METHODS: Transiently evoked otoacoustic emissions (TEOAEs) and Distortion-product otoacoustic emissions(DPOAEs) of 28 patients with OSAHS were measured one week before uvulopalatopharyngoplasty (UPPP) and six months after UPPP. RESULTS: In these patients the prevalence of TEOAEs was 64.3%, its spectral energy was mainly distributed between 0.5-2 kHz, the amplitudes of DPOAEs between 0.5-8 kHz were reduced significantly, the prevalences of DPOAEs at 0.5-8 kHz were between 46.4%-89.3%. After UPPP, the prevalences of TEOAEs and DPOAEs were increased, the amplitudes of DPOAEs were in turn (22.50 +/- 16.70) dB SPL, (5.83 +/- 2.69) dB SPL, (0.69 +/- 3.83) dB SPL, ( 2.50 +/- 2.45) dB SPL, (4.95 +/- 2.65) dB SPL, (4.48 +/- 3.07) dB SPL from 0.5 kHz to 8 kHz, the amplitudes of DPOAEs were significantly increased at 1 kHz, 2 kHz, 4 kHz, 6 kHz, 8 kHz. CONCLUSION: OSAHS can affect the cochlear function of the patients with OSAHS, UPPP can improve the cochlear function. PMID- 15127571 TI - [Protective effect of ONO-1078, a leukotriene receptor antagonist, on focal cerebral ischemia induced by endothelin-1 in rats]. AB - AIM: To determine the protective effect of ONO-1078, a leukotriene receptor antagonist, on focal cerebral ischemia induced by endothelin-1 in rats. METHODS: Slow microinjection of endothelin-1 (120 pmol in 6 microL, for > 6 min) into the region near the middle cerebral artery was used to induce focal cerebral ischemia. ONO-1078 (0.1 mg.kg-1) was i.p. injected 1 h before endothelin-1 injection. Neurological symptoms, brain edema, brain infarction size, and the survival neurons in cortex and striatum were observed 24 h after ischemia. RESULTS: Intracerebral microinjection of endothelin-1 induced remarkable neurological symptoms, brain infarction, brain edema, and decrease of survival neurons in the cortex and striatum. In rats pretreated with ONO-1078, endothelin 1-induced brain edema and brain infarction size were decreased. The numbers of survival neurons in striatum and cortex were increased significantly. The neurological symptoms were improved but not significantly. CONCLUSION: ONO-1078 possesses neuroprotective effect against cerebral ischemic injury induced by endothelin-1, therefore, leukotrienes may play a role in the injury of cerebral ischemia. PMID- 15127572 TI - [Effects of emodin on the intracellular calcium concentration ([Ca2+]i) and L type calcium current of the single ventricular mytocytes from guinea pig]. AB - AIM: To study the effects of emodin on intracellular calcium concentration ([Ca2+]i) and L-type calcium current of the single ventricular myocytes from guinea pig. METHODS: Enzymatic dissociation was used to isolate single ventricular myocytes from adult guinea pig. They were loaded with Ca2(+) sensitive fluorecent indicator Fluo-3/AM. [Ca2+]i represented by fluorescent intensity (FI) was measured by laser scanning confocal microscope. Whole cell patch clamp technique was used to record ICa-L. RESULTS: At resting status, [Ca2+]i was not affected by emodin (1-100 mumol.L-1). Emodin at the concentration of 1 mumol.L-1 was shown to increase the [Ca2+]i induced by 60 mmol.L-1 KCl. The peak value of fluorescent intensity was increased from 1,877 +/- 551 to 2,905 +/- 739 (n = 8, P < 0.05). Emodin at the concentration of 10 mumol.L-1 had no effect on the increase of [Ca2+]i induced by 60 mmol.L-1 KCl. However, the increase of [Ca2+]i induced by KCl was reduced to 1,214 +/- 335 (n = 8, P < 0.05) by 100 mumol.L-1 emodin. The density of ICa-L was increased from (-6.2 +/- 1.3) pA/pF to (-8.3 +/- 0.3) pA/pF (n = 6, P < 0.05) by 1 mumol.L-1 emodin at the test pulse of 0 mV. The current was not altered by 10 mumol.L-1 emodin. But it was inhibited from (-6.6 +/- 1.0) pA/pF to (-3.80 +/- 0.16) pA/pF (n = 6, P < 0.05) by 100 mumol.L-1 emodin at the test pulse of +10 mV. CONCLUSION: Emodin has two-way regulation on [Ca2+]i and ICa-L of cardiomyocytes in guinea pig. PMID- 15127573 TI - [Inhibition of voltage-gated K+ current in rat intrapulmonary arterial smooth muscle cells by endothelin-1]. AB - AIM: To investigate the role of endothelin-1 (ET-1) in the physiological and pathophysiological regulating mechanisms of voltage-gated K+ current (IKV) inhibition in rat intrapulmonary arterial smooth muscle cells (PASMCs). METHODS: Single PASMCs were obtained with acute enzyme (collagnase plus papain) dispersing method. Using whole cell patch-clamp technique in freshly isolated rat PASMCs, the effect of ET-1 on voltage-gated K+ current was recorded. RESULTS: ET-1 (1 x 10(-9) mol.L-1) and the voltage-dependent K+ (KV)-channel antagonist 4 aminopyridine (4AP), but not the Ca(2+)-activated K(+)-channel antagonist tetraethylammonium (TEA), caused membrane depolarization. The effect of ET-1 on membrane potential persisted in cells in which intracellular Ca2+ was buffered with 1,2-bis (2-aminophenoxy) ethane-N,N,N',N'-tetraacetic acid (BAPTA). ET-1 (1 x 10(-9) mol.L-1) caused a significant reversible inhibition of KV current, which began 4.0 s after application of ET-1, was much earlier than the effect of membrane depolarization of PASMCs (15s). ET-1 (1 x 10(-10) to 1 x 10(-7) mol.L-1) caused a concentration-dependent inhibition of K+ current ( mV, from 136 to 40 pA/pF). The percent inhibition was 71% at 1 x 10(-7) mol.L-1 (n = 6). The effect of ET-1 (1 x 10(-9) mol.L-1) on K+ current was weaker under conditions free of Ca2+ than containing Ca2+. At a test potential of mV, free of Ca2+ conditions reduced the maximum inhibitory effect of ET-1 from 71% to 50%. CONCLUSION: ET-1 modulated pulmonary vascular reactivity by depolarizing membrane potential and inhibiting the K+ current of PASMCs. The effect of ET-1 on PASMCs membrane potential and the inhibition of K+ current were independent of Ca2+, but the inhibition of K+ current was much greater under conditions containing Ca2+ than free of Ca2+. PMID- 15127574 TI - [Molecular mechanisms of antioxidant effects of propylene glycol mannate sulfate]. AB - AIM: To investigate the antioxidant mechanisms of propylene glycol mannate sulfate (PGMS) in hyperlipidemic rats. METHODS: Male Wistar rats were given high lipid emulsion diet to establish hyperlipidemic model. PGMS was given every day at different doses (37.8 and 75.6 mg.kg-1, ig) to hyperlipidemic rats for three weeks. In addition, diethyldithiocarbamate (DDC) was given 200 mg.kg-1.3 d-1 (i.p.) to inhibit SOD activity. Then, the MDA content was examined using TBA method to show the oxidation level, and the activities of SOD, GSH-Px and CAT were examined following the kit protocols to indicate the capability of eliminating OFR. RT-PCR was applied to study the expression of Cu, Zn-SOD mRNA in rat liver. RESULTS: The MDA content of PGMS treatment groups decreased markedly compared with hyperlipidemic group, and the activities of SOD, GSH-Px and CAT increased distinctly. Cu, Zn-SOD mRNA expression was significantly increased by PGMS treatment. Furthermore, the application of DDC(the SOD inhibitor) reduced total SOD activity and Cu, Zn-SOD mRNA expression induced by PGMS, and the content of MDA increased correspondingly. CONCLUSION: PGMS can induce the activities of antioxidant enzymes and the mRNA expression of Cu, Zn-SOD, which contribute to the elimination of oxygen free radical. This may explain the molecular mechanism of antioxidant effects of PGMS. PMID- 15127575 TI - [Protective effects of shark hepatic stimulator substance against acute hepatic injury induced by acetaminophen in mice]. AB - AIM: To investigate the protective effects of shark hepatic stimulator substance (sHSS) against acute hepatic injury induced by acetaminophen (AAP) in mice. METHODS: Acute hepatic injury model of Balb/c mice was induced by a single intraperitoneal injection of AAP (200 mg.kg-1, i.p.). Serum ALT and AST activities were analyzed. The changes of microstructure and ultrastructure of hepatocyte were observed under optical and electronic microscope. The hepatocyte apoptosis was analyzed by flow cytometer and the expression level of Fas mRNA was determined by RT-PCR. RESULTS: The activities of serum ALT and AST were significantly decreased and both necrosis and inflammatory infiltration were improved in the mice treated with sHSS 3.0 and 1.5 mg.kg-1. sHSS (3.0 mg.kg-1) prevented the ultrastructural changes of hepatocytes caused by AAP, decreased the percentage of apoptotic cells, and downregulated the expression level of Fas mRNA. CONCLUSION: sHSS protected hepatocytes from AAP-induced injury, which might be associated with its protection of the mitochondria and inhibition of apoptosis and expression of Fas mRNA in hepatocytes. PMID- 15127576 TI - Molecular mechanism of granulocytic differentiation of human promyelocytic leukemia HL-60 cells induced by all-trans retinoic acid. AB - AIM: To elucidate the molecular mechanism of granulocytic differentiation of human promyelocytic leukemia HL-60 cells induced by all-trans-retinoic acid (ATRA). METHODS: Flow cytometry was used to determine the cell cycle changes of HL-60 cells upon ATRA treatment. Gene expression profiles of HL-60 cells induced by 1 mumol.L-1 ATRA were obtained by using cDNA microarrays containing 9,984 genes and expressed sequence tags (ESTs). RESULTS: Cell cycle analysis showed that 48%-73% of cells were arrested at G1/G0 phase upon ATRA treatment; cDNA microarray results demonstrated that the expression of genes encoding adhesion molecules, tissue remodeling proteins, transporters and ribosomal proteins were up-regulated in ATRA treated of HL-60 cells. Several genes involved in oxidase activation pathway were also differentially expressed. CONCLUSION: ATRA treatment induced growth arrest and differentiation in HL-60 cells, which is associated with regulation of the oxidase activation pathway and the expression of tissue remodeling proteins. PMID- 15127577 TI - [Inhibitory effects of artesunate on angiogenesis]. AB - AIM: To investigate the inhibitory effects of artesunate on angiogenesis. METHODS: The in vitro anti-angiogenic effect of artesunate was tested on models of angiogenesis: proliferation, migration and tube formation of human umbilical vein endothelial (HUVE) cells; The anti-angiogenic effect in vivo was evaluated in nude mice by means of human ovarian cancer HO-8910 implantation and immunohistochemical stainings for microvessel (CD31), vascular endothelial growth factor (VEGF) and VEGF receptor KDR/flk-1. RESULTS: Artesunate significantly inhibited angiogenesis in a concentration-dependent form in range of 0.5-50 mumol.L-1. The IC50 of artesunate for HUVE cells was (21 +/- 3) mumol.L-1. Growth of xenograft tumor was decreased and microvessel density was reduced following drug-treatment with no apparent toxicity to the animals. Artesunate also remarkably lowered VEGF expression on tumor cells and KDR/flk-1 expression on endothelial cells as well as tumor cells. CONCLUSION: Artesunate was shown to inhibit angiogenesis in vitro and in vivo. These findings together with the known low toxicity of artesunate are clues that artesunate may be a promising angiogenesis inhibitor. PMID- 15127578 TI - [Effects of (-), (+)-7-hydroxy-clausenamide on synaptic transmission in rat dentate gyrus in vivo]. AB - AIM: To investigate effects of (-), (+)-7-hydroxy-clausenamide (7-OH-Clau) on basal synaptic transmission in the dentate gyrus of anesthetized rats. METHODS: Extracellular recording of the population spike (PS) of hippocampal dentate gyrus following application of low-frequency stimulation (1/30 Hz, 1.0 mA). RESULTS: Fifteen, thirty and sixty minutes after intracerebroventricular (icv) administration of an estimated final brain concentration of 2 x 10(-6) mol.L-1, ( )-7-OH-Clau caused over 30% increase relative to basal PS amplitude before administration, and 27%-41% increase relative to vehicle control (P < 0.05), while (+)-7-OH-Clau exhibited 18%-25% and 11%-20% decrease in the PS amplitude when compared with basal PS amplitude and vehicle control (P < 0.05), respectively. CONCLUSION: The basal synaptic transmission in the dentate gyrus of the rat hippocampus can be potentiated by (-)-7-OH-Clau and attenuated by (+)-7 OH-Clau, indicating that there was a stereoselective difference between the two enantiomers in the modulation of synaptic responses and plasticity. PMID- 15127579 TI - Synthesis and antioxidative activity of 2-substituted phenyl-5-(3'-indolyl) oxazole derivatives. AB - AIM: To study the synthesis of 5-(3'-indolyl)-oxazoles and their antioxidative activity. METHODS: The amides were prepared from tryptophan and different acid derivatives by the catalytic dehydration of dicyclohexyl carbodiimide (DCC). The characteristic heterocyclic ring system of 5-(3'-indolyl)-oxazoles was constructed by oxidative cyclization of amide, using dicholorodicyanoquinone (DDQ). Their antioxidative activity in vitro was tested using DPPH system. RESULTS: Eleven 2-substituted phenyl-5-(3'-indolyl)-oxazoles were prepared, the compounds 21 and 22 have shown antioxidative activity 3-4 times stronger than that of Vit E, and the compound 29 showed antioxidative activity almost as same as Vit E. CONCLUSION: Three 5-(3'-indoyl)-oxazole compounds synthesized showed potent antioxidative effect and they would be a good antioxidants. PMID- 15127580 TI - [Isolation and structural identification of chemical constituents from Selaginella tamariscina (Beauv.) Spring]. AB - AIM: To study the chemical constituents of the water-extracts of Selaginella tamariscina (Beauv.) Spring. METHODS: Various chromatographic techniques were used to separate and purify the constituents. Their physico-chemical properties and spectral data were used to elucidate the structures. RESULTS: Nine compounds were isolated and identified as (2R,3S)-dihydro-2- (3',5'-dimethoxy-4' hydroxyphenyl)-7-methoxy-5-acetyl-benzofuran (1), 3-hydroxy-phenpropionic acid (2'-methoxy-4'-carboxy-phenol) ester (tamariscina ester A, 2), sygringaresinol (3), 1-(4'-hydroxyl-3'-methoxyphenyl)glycerol (4), ferulic acid (5), caffeic acid (6), vanillic acid (7), syringic acid (8), and umbelliferone (9). CONCLUSION: Compound 1 and 2 are new compounds, and the others were isolated from Selaginella for the first time. PMID- 15127581 TI - [Isolation and structure identification of chemical constituents from the seeds of Descurainia sophia (L.) Webb ex Prantl]. AB - AIM: To isolate and determine the structures of chemical constituents from the seeds of Descurainia sophia (L.) Webb ex Prantl. METHODS: The chemical constituents were extracted from the seeds of Descurainia sophia (L.) Webb ex Prantl with 75% ethanol and purified by polyamide, silica gel, RP-C18 and Sephadex LH-20 on column chromatography. Chemical methods and spectroscopic methods, such as 1H and 13CNMR, HSQC, HMBC and TOCSY spectra were used for the structural identification. RESULTS: Fifteen compounds were obtained. Twelve of them were identified as quercetin-3-O-beta-D-glucopyranosyl-7-O-beta gentiobioside (I), kaempferol-3-O-beta-D-glucopyranosyl-7-O-beta-gentiobioside (II), isorhamnetin-3-O-beta-D-glucopyranosyl-7-O-beta-gentiobioside (III), quercetin-7-O-beta-gentiobioside (IV), kaempferol-7-O-beta-gentiobioside (V), isorhamnetin-7-O-beta-gentiobioside (VI), quercetin-3,7-di-O-beta-D glucopyranoside (VII), kaempferol-3, 7-di-O-beta-D-glucopyranoside (VIII), isorhamnetin-3, 7-di-O-beta-D-glucopyranoside (IX), kaempferol-3-O-beta-D glucopyranosyl-7-O-[(2-O-trans-sinnapoyl)-beta-D- glucopyranosyl(1-->6)]-beta-D glucopyranoside) (X), sinapic acid ethyl ester (XI) and 3, 4, 5-trimethoxyl cinnamic acid (XII). CONCLUSION: Compounds X and VI are new compounds. IV, V, VII, VIII and IX were isolated from Cruciferae family for the first time. I, II, III were obtained from Descurania genus and XI, XII from D. sophia for the first time. PMID- 15127582 TI - [Isolation and purification of gonyautoxins from Alexandrium mimutum Halim]. AB - AIM: To isolate and purify gonyautoxins from Alexandrium mimutum Halim Amtk2 strain. METHODS: The ethanol extracts of culture Alexandriun minutum Halim Amtk2 were isolated by means of gel filtration chromatography, the toxin fraction obtained was then purified by ion exchange chromatography. RESULTS: From 100 liter of cultivation liquid of Alexandrium mimutum Halim Amtk2 (6.74 +/- 0.31) x 10(9) cells were obtained. The ethanol extracts of Alexandriun minutum Halim purified by gel filtration chromatography obtained gonyautoxins mixture 29.59 mg. 4.06 mg of the mixture was further purified by two steps of ion exchange chromatography, and obtained pure GTX-4 (0.40 +/- 0.002) mg, GTX-1 (5.95 +/- 0.03) x 10(-2) mg, GTX-3 (6.92 +/- 0.05) x 10(-4) mg and GTX-2 (0.11 +/- 0.005) mg. CONCLUSION: Pure gonyautoxins can be obtained by means of gel filtration chromatography and ion exchange chromatography from ethanol extracts of cultured Alexandriun minutum Halim Amtk2 strain. PMID- 15127583 TI - Simultaneous determination of the major isosteroidal alkaloids and their glucosides in the bulbs of Fritillaria by high performance liquid chromatography coupled with evaporative light scattering detection. AB - AIM: To establish an HPLC-ELSD method for the simultaneous determination of five major bioactive isosteroidal alkaloids and gluco-alkaloids in the bulbs of Fritillaria namely peimissine, imperialine, sinpeinine A, imperialine-3 beta glucoside and yibeinoside A. METHODS: A Nova-Pak C18 column (150 mm x 3.9 mm ID) was used. The chromatography was carried out with a linear gradient programming. The mobile phase was acetonitrile-water (containing 0.1% diethylamine) and the flow rate was 1.0 mL.min-1. RESULTS: The linear range of peimissine was 13.1 288.2 mg.L-1 (r2 = 0.9975), imperialine-3 beta-glucoside 7.7-169.4 mg.L-1 (r2 = 0.9993), yibeinoside A 7.3-160.6 mg.L-1 (r2 = 0.9997), imperialine 16.5-363.0 mg.L-1 (r2 = 0.9992), sinpeinine A 8.7-191.4 mg.L-1 (r2 = 0.9942). CONCLUSION: The method is accurate with overall intra- and inter-day variation less than 5% and recovery more than 95%. The method was successfully applied to analyze five major bioactive alkaloids and gluco-alkaloids in three Fritillaria bulbs. PMID- 15127584 TI - [A near-infrared diffuse reflectance analysis method for the noninvasive quantitative analysis of ambroxol hydrochloride tablets]. AB - AIM: To develop a near-infrared diffuse reflectance analysis (NIRDRA) method for rapid noninvasive quantitative determination of ambroxol hydrochloride in half finished product particles and non-blister-packed, blistered tablets. METHODS: All spectra were measured with a Fourier transform spectrometer equipped with a PbS and a InGaAs detector, an external integrating sphere, a rotating sample cup, and a fibre-optic probe for reflectance measurements. All samples were scanned from 12,000 cm-1 to 4,000 cm-1, and each sample spectrum was obtained as an automatic mean of 64 scans. No spectrum pre-processing method was used, and spectral regions, 4,602-4,247, 12,000-7,498 and 6,102-5,446, 12,000-5,446 cm-1 were selected to develope mathematical models by partial least square method for half-finished product particles and non-blister-packed, blistered tablets samples, respectively. RESULTS: The optimal rank and mean square error determined for half-finished product particles and non-blister-packed, blistered tablets samples by cross validation method all was 6 and 0.306, 0.972 and 1.492, respectively, the average recovery was 100%, 100% and 102% respectively; and the RSD was 1.17%, 1.70% and 1.78% respectively. CONCLUSION: Results showed that the NIRDRA method was rapid, simple, noninvasive and sensitive, and it can be applied to assay the content of ambroxol hydrochloride in half-finished product particles non-blister-packed and blistered tablets. PMID- 15127585 TI - Monitoring sub-nanogram amount of acetylspiramycin in human urine using flow injection analysis with chemiluminescence detection. AB - AIM: To establish a new and simple flow injection method for the rapid determination of acetylspiramycin (ASPM). METHODS: ASPM was determined by chemiluminescence (CL) method combined with flow injection (FI) technology, which was based on the inhibitive effect of ASPM on the chemiluminescence reaction of the luminol-K3Fe (CN)6 system. RESULTS: The decrease of chemiluminescence intensity was proportional to the logarithm of ASPM concentration (0.1-100) microgram.L-1, the detection limit was 40 ng.L-1 (3 sigma). The whole process, including sampling and washing, could be completed in 0.5 min with a RSD less than 3.0% (n = 5). CONCLUSION: The FI-CL method is of both high sensitivity and good selectivity giving a throughput of 120 h-1. The proposed method was applied successfully to the determination of ASPM in pharmaceutical preparations and human urine without any pre-treatment. It was found that the ASPM concentration reached its maximum after being orally administrated for two hours. PMID- 15127586 TI - [Study on preparation conditions for polylactide nanoparticles loaded cyclosporine A and its oral bioavailability in rats]. AB - AIM: To develop a less toxic alternative for sandimmun neoral (Neoral). To study the preparation conditions and to compare its pharmacokinetic characteristics with Neoral. METHODS: Polylactide nanoparticles loaded cyclosporine A was prepared by solvent-nonsolvent method. Polylactide nanoparticles were administered by oral in a dosage of 15 mg.kg-1. The CyA concentration in whole blood sample was determined by HPLC. RESULTS: The quantities of CyA, PLA and volume of acetone added had significant influence on the NP diameters. Under proper condition, the nanoparticles with diameters of 57.5 nm were obtained. The relative bioavailability in rats was 101.6%, with a smaller absorption rate (P < 0.05) and a smaller elimination rate (P < 0.1). CONCLUSION: The nanoparticles (diamater < 100 nm) with relative high bioavailability were prepared using solvent-nonsolvent method. It is suitable for further study. PMID- 15127587 TI - [Preparation of cationic liposomes and its role in enhancing cellular uptake of antisense oligonucleotides]. AB - AIM: To prepare the liposomes which protect antisense oligodeoxynucleotides (ASON) against nuclease degradation and delivery ASON into cytoplasmic efficiently. METHODS: A cationic derivative of cholesterol, 3 beta-[N-(N',N' dimethylaminoethan)-carbamoyl] cholesterol (DC-Chol) was synthesized and used to prepare cationic liposome. The characteristics of liposomes/ASON complexes including size, drug loaded efficiency and structure were investigated. Cellular uptake of fluorescence labled ASON (FAM-ASON) under different condition was determined by flow cytometric analysis. Denatured polyacryamide gel electrophoresis (DPGE) was used to analyze the role of liposomes in protecting ASON. RESULTS: The mean values of preliposomes and liposomes/ASON complexes size were 185.7 and 228.2 nm, respectively. Cationic liposomes showed a high adsorption capacity for ASON. When the +/- charge ratio exceeded 2:1, more than 90% of the ASON was loaded into liposomes. Agarose gel electrophoresis showed three different existence of ASON in liposomes formulation: free, absorbed and encapsulated types. Concerning cellular uptake, DC-Chol liposomes indicated high efficient effect of increasing cellular uptake of ASON. Compared with free ASON, the total fluorescence intensity in cytoplasma was significantly enhanced. The level of increasing was largely depended on +/- charge ratio. The cellular uptake of FAM-ASON decreased in the presence of serum. The cellular total fluorescence intensity in 10% and 30% fetal bovine serum of cultured medium were only 22.3% and 15.5% as that of serum-free media, respectively. DPGE confirmed that free ASON was rapidly degraded by DNase I while ASON encapsulated into liposomes was efficiently protected. CONCLUSION: The cationic DC-Chol liposomes are shown to be promising carriers to deliver ASON into cytoplasma. PMID- 15127588 TI - [Absorption of recombinant hirudin in rats GI tract]. AB - AIM: To investigate the absorption and distribution of recombinant hirudin-2 (rHV2) in the GI tract in rats after oral administration. METHODS: Using HPLC, fluorescence spectrophotometry and confocal laser scanning microscopy to measure the amount of intact rHV2 absorbed into gastrointestinal mucosa and blood. RESULTS: HPLC spectrum showed that there were intact rHV2 molecules in plasma after 1 h of oral administration. After oral administration for 3 h, 1.2%-26.8% of fluorescence was found in the GI tract in rats. Chromatographic analysis showed that 2.27%-38.75% of fluorescence recovered from the GI tract luminal contents and mucosa was eluted at the void volume of a Sephadex G-25 column. Microscopic examination showed that FITC-rHV2 was taken up throughout the whole small intestine but the ileum appeared to be a preferred site for FITC-rHV2 transport in rats. CONCLUSION: rHV2 may partially survive in the GI lumen and subsequently absorbed in active form by gastrointestinal tract. PMID- 15127589 TI - [Insisting on- to get success--my experience in the study of herbal pharmacology]. PMID- 15127590 TI - [Adipocyte differentiation and its regulation]. AB - Fat tissue and adipocytes have been exclusively investigated in the past two decades, especially in the last ten years, due to the following two reasons. Firstly, more and more studies showed that fat tissue is not only an organ for energy storage, but also an endocrine one that can secret many kinds of hormones or hormone-like peptides. Secondly, the established preadipocyte cell lines have been providing powerful tools for the in vitro research of adipocyte differentiation, because these immortal cell lines authentically represent, to a great extend, the in vivo situations of these cells and can be induced to differentiate into mature adipose cells with proper hormones. It has been demonstrated that there exists close relations between adipocyte differentiation and many physiological or pathological processes including saccharide and fat metabolism, energy balance, obesity, diabetes, hyperlipidemia and breast cancer. It is very important to make known the differentiation mechanisms of preadipocyte into adipocyte for understanding the above mentioned diseases and for screening anti-obesity and anti-diabetes drugs. PMID- 15127591 TI - [Peroxisome proliferator-activated receptors and atherosclerosis]. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-dependant nuclear transcription factors, consisting of three isoforms: alpha,beta/delta and gamma, which form a subfamily of the nuclear receptors superfamily. PPARs play an important role in adipocyte differentiation, energy metabolism, and inflammation. PPARs' effect goes beyond the improvement of insulin resistance in syndrome X (diabetes mellitus, hypertension, obesity and so on). Through recent years' study, it has been demonstrated that PPARs regulate vascular wall directly, which, therefore, decelerate the development of atherosclerosis. In this review, we will look at current trends of PPARs research in their structure, function, and molecular mechanism related to pathogenesis and therapy of atherosclerosis. PMID- 15127592 TI - [Progress in the study of pain-related somatosensory evoked potentials]. AB - Important progresses has been made in the study of pain-related somatosensory evoked potentials (pain SEP) in the past decades. Pain generating stimulation techniques in humans are used in combination with the methodology of evoked electrical brain potentials. The current review will discuss stimulation techniques, components of evoked potentials, dipole source analysis of evoked potentials with the recent study of pain SEP. PMID- 15127593 TI - [Research progress on regulation of class II MHC expression]. AB - Class II MHC antigens play a critical role in the induction of immune responses through presentation of processed antigen to CD4+ T lymphocytes. The absence of MHC II normal expression results in severe primary immunodeficiency diseases, such as the bare lymphocyte syndrome (BLS). Four different MHC II regulatory genes have been identified. These genes encode RFXANK, RFX5, RFXAP and CIITA. The first three are subunits of RFX, a ubiquitously expressed factor that binds to the promoters of all MHC II genes. CIITA is the master control factor for MHC II expression. The highly regulated expression pattern of CIITA ultimately dictates the cell type specificity, induction and level of MHC II expression. This review focuses on research progress on regulation of class II MHC expression in recent years. PMID- 15127594 TI - [Progress in gene therapy of liver cirrhosis]. AB - Liver cirrhosis is a common progressive pathological lesion in the late stage of chronic liver disease, which is characterized by disorganization of normal hepatic structure of regenerative nodules and hyperplasia of fibrotic tissues. In recent years, with the development of molecular biology, the molecular mechanisms underlying liver cirrhosis has been revealed more and more, which makes the therapy at the gene level possible. The ideal strategy for the treatment of liver cirrhosis should include prevention of fibrogenesis, stimulation of hepatocyte proliferation and reorganization of the liver architecture. Several gene therapy approaches for treatment of liver cirrhosis have been developed by transfer of some genes of cytokines and enzymes (e.g. HGF, TGF beta 1R, MMPs). These gene therapies can inhibit fibrogenesis and hepatocyte apoptosis and also produce resolution of fibrosis in the cirrhotic liver. Thus, gene therapy may be potentially useful for the treatment of liver cirrhosis, which is otherwise fatal and untreatable by conventional therapy. PMID- 15127595 TI - [Current progress of matrix metalloproteinase in acute lung injury following cardiopulmonary bypass]. AB - Degradation of basement membrane of pulmonary capillaries was the main pathological feature of lung injury following cardiopulmonary bypass (CPB). Matrix metalloproteinases (MMPs) contributed to the development of lung injury following CPB via degrading extracellular matrix and regulating cytokines. So, understanding the effects of MMPs in the lung injury following CPB may be helpful to prevent and treat the complications mentioned above. PMID- 15127596 TI - [Update on oligodendrocyte in biological function and associated neurological disorders]. PMID- 15127597 TI - [Mechanism of migration of neural precursor cells and it's application]. PMID- 15127598 TI - [CDK5 and neurodegenerative disease]. PMID- 15127599 TI - [A new serine protease with limited distribution specially in heart--Corin]. PMID- 15127600 TI - [The advance of RNA interference]. PMID- 15127601 TI - [Recent advances in study on stimulator of Fe transport]. PMID- 15127602 TI - [The role of HOXB4 gene in the regulation of proliferation and differentiation of hematopietic stem cell]. PMID- 15127604 TI - [Homocysteine and Alzheimer's disease]. PMID- 15127603 TI - [Correlation of HIV and herpesvirus infections]. PMID- 15127605 TI - [Regulation of lipid metabolism by liver X-activated receptors]. PMID- 15127606 TI - [The effects of metabotropic glutamate receptors in synaptic plasticity]. PMID- 15127607 TI - [Regulation of neuronal L-type voltage-gated calcium channels and brain ischemia]. PMID- 15127608 TI - [Progress in the study of cyclooxygenase]. PMID- 15127609 TI - [Physiological and pathophysiological roles of perlecan]. PMID- 15127610 TI - [Effects of edaravone, minocycline and ONO-1078 on oxygen/glucose deprivation induced electrophysiological alteration in rat hippocampal slices]. AB - AIM: To establish an in vitro model of hippocampal slice to detect electrophysiological alteration after oxygen/glucose deprivation (OGD), and to observe the effects of edaravone, minocycline and ONO-1078 [pranlukast, 4-oxo-8 [p-(4-phenylbutyloxy) benzoyl-amino]-2-(tetrazol-5-yl)-4H-1-benzopyran hemihydrate]. METHODS: Hippocampal slices from rats were perfused with artificial cerebrospinal fluid lacking oxygen and glucose for 3, 4, 7 and 10 min. The population spike (PS) was recorded, and 2,3,5-triphenyltetrazolium chloride (TTC) staining was performed in some experiments, to detect the slice viability in the presence or absence of drugs in the perfusion solution. RESULTS: Four min of OGD treatment was the most suitable duration for induction of slice injury, and PS amplitudes were recovered to (29 +/- 6)% of baseline values within 1 h after 4 min OGD. Edaravone, a free radical scavenger, at 1 and 10 mumol.L-1 significantly increased the recovery rate to (56 +/- 13)% and (69 +/- 12)% of baseline respectively 1 h after OGD. However, the anti-inflammatory drug minocycline (10 mumol.L-1) and leukotriene receptor antagonist ONO-1078 (1 mumol.L-1) did not increase the recovery. NMDA receptor antagonist ketamine, as a positive control, also promoted the recovery concentration-dependently. CONCLUSION: OGD for 4 min was a feasible in vitro ischemia model for determination on electrophysiological alteration in hippocampal slices. Edaravone showed concentration-dependent protective effect on OGD injury, and anti-inflammatory drugs minocycline and ONO 1078 showed no effect. PMID- 15127611 TI - [High throughput screening method of potassium channel regulators]. AB - AIM: To discover new regulators of potassium channel, an in vitro assay based on DiBAC4 (3) to determine the fluorescence was established for high throughput screening. METHODS: A cell-based 96-well format fluorescence assay using DiBAC4 (3) in cultured PC12 cells was described. Cells were loaded with 5 mumol.L-1 DiBAC4 (3) and incubated at 37 degrees C for 30 min before adding KCl or several known potassium channel regulators. The cellular DiBAC4 (3) fluorescence responce was then detected. The fluorescence changes can be used to evaluate membrane potential changes, which are determined mainly by potassium channels. RESULTS: Extracellular high K(+)-induced depolarization and several potassium channel blockers including 4-AP, TEA, E-4031, glibenclamide, quinidine and nifedipine all evoked increases in DiBAC4 (3) fluorescence response. The potassium channel opener, cromakalim, evoked decrease in DiBAC4 (3) fluorescence response. The fluorescence changes of 4-AP, TEA, glibenclamide, nifedipine and cromakalim were in a concentration-dependent manner. In 76 compounds screened by using the established DiBAC4 (3)-based assay, 9 compounds were found to change the fluorescence dose-dependently. Patch clamp technique is needed to further testify and screen their actions on potassium currents. CONCLUSION: The DiBAC4 (3)-based assay is easily operated, economical and repeatable. So, it can be performed by high throughput screening for potassium channel regulators. PMID- 15127612 TI - Effect of combination of dizocilpine with general antiepileptic drugs on amygdala kindling models in rats. AB - AIM: To investigate the antiepileptic effect of dizocilpine (MK-801) on amygdala kindling models in rats and the effects of its combination with general antiepileptic drugs. METHODS: To establish amygdala kindling models in rats and observe the effect of dizocilpine on kindling models and its combination with general antiepileptic drugs (phenobarbital, valproate and nicardipine) at ineffective dose. The influence of dizocilpine on convulsions induced by semicarbazide (SCZ) in mice were also observed. RESULTS: Dizocilpine (0.1-0.25 mg.kg-1, i.p.) was shown to dose-dependently inhibit amygdala kindled seizure, shorten the after discharge duration (ADD) and reduce the Racine's stage (P < 0.01). The combination of dizocilpine with phenobarbital, valproate, nicardipine at ineffective dose shortened ADD or reduced Racine's stages (P < 0.01). Dizocilpine (0.1-0.25 mg.kg-1, i.p.) significantly prolonged the latency and reduced the rate of convulsions and death in mice. CONCLUSION: Dizocilpine inhibits the seizure of the amygdala kindling and improve the antiepileptic activity of phenobarbital, valproate and nicardipine, indicating that these combination may provide a new approach for treating epilepsy. PMID- 15127613 TI - [Inhibition of tacrine on delayed rectifier and transient outward potassium currents in cultured rat hippocampal neurons]. AB - AIM: To study the effects of tacrine on IK and IA potassium current in primary cultured rat hippocampal neurons. METHODS: Whole cell patch clamp and primary rat hippocampal neuron cultures were used. RESULTS: Tacrine was shown to reduce the amplitude of IK and IA, in concentration-dependent manners. The IC50s at +40 mV for reduction of IK and IA were 23 and 52.6 mumol.L-1, respectively. Tacrine (30 mumol.L-1) shifted the steady state activation of IK and IA to negative potentials by 12 and 15 mV, respectively. The V1/2 of activation curves for IK current before and after the application of tacrine were (6.7 +/- 1.4) mV and ( 5.4 +/- 1.3) mV, respectively. The k of activation curves for IK current was 13.4 + 1.3 and 12.5 + 1.4 without and with tacrine, respectively. The V1/2 of activation curve for IA current were (-9.9 +/- 2.6) mV and (-24 +/- 5) mV in the absence and presence of tacrine, respectively, and the k value was not changed. CONCLUSION: Tacrine inhibited IK and IA currents in rat hippocampal neurons and it is more potent for blocking IK. PMID- 15127614 TI - [Effects of 9-cis-retinoic acid on cell cycle and expression of cyclin D1 and cdk4 in lung cancer cells]. AB - AIM: To study the effects of 9-cis-retinoic acid (9-cis-RA) on cell cycle and expression of cyclin D1 and cdk4 in lung cancer cells. METHODS: 9-cis-RA (1 x 10( 6) mol.L-1) was used to treat lung cancer cells for 24 h; Flow cytometry (FCM) was used to detect the percent of G0/G1 phase and S phase cells of three groups including blank control, DMSO control and 9-cis-RA groups; RT-PCR was used to analyze the expression changes of cyclin D1 and cdk4 before and after treatment with 9-cis-RA in lung cancer cells. RESULTS: The percent of G0/G1 phase cells of 9-cis-RA groups was significantly higher than that of the control groups (P < 0.01 or P < 0.05) and the percent of S phase cells of 9-cis-RA groups was lower than that of the control groups (P < 0.01 or P < 0.05); the expression of cyclin D1 of PG, SPC-A1 and L78 cells was decreased (P < 0.01) and the expression of cdk4 of PG, A549 and L78 cells was also decreased (P < 0.01) after treatment with 9-cis-RA. CONCLUSION: Most of the proliferation and the expression of cyclin D1 and cdk4 of PG, A549, SPC-A1 and L78 were inhibited by 9-cis-RA. PMID- 15127615 TI - [Activation of Ca(2+)-activated K+ channels by oxyphenamone in rabbit mesenteric vascular smooth muscle cells]. AB - AIM: To study the effects of oxyphenamone (Oxy) on activation of Ca(2+)-activated K+ channels in rabbit mesenteric vascular smooth muscle cells. METHODS: To measure the effect of Oxy on the Ca(2+)-activated K+ channel (BK (Ca) channel) activity in rabbit mesenteric vascular smooth muscle cells by using whole cell patch clamp techniques. RESULTS: Oxy reversibly increase BK (Ca) channel activity in rabbit mesenteric artery smooth muscle cells. Application of Oxy (0.1 mumol.L 1) to the perfusion solution caused significant increase in outward currents and its effect was completely abolished by washout; The outward currents K+ was inhibited by TEA (7.5 mmol.L-1); Oxy activated the BK (Ca) channel in a dose dependent manner (0.01-10 mumol.L-1). CONCLUSION: Oxy directly increase the activity of BK (Ca) channel activity in rabbit mesenteric vascular smooth muscle cells in dose-dependent manner. PMID- 15127616 TI - [Synthesis of (+/-) ibuprofen sugar derivatives]. AB - AIM: (+/-) Ibuprofen sugar derivatives were prepared in order to decrease side effects and increase bioavailability of (+/-) ibuprofen. METHODS: The synthesis of derivatives were performed using 1,2:3, 4-di-O-isopropylidene-beta-D galactopyranose, 1,3,4,5-tetra-O-acetyl-2-deoxy-2-amino-beta-D-gluctopyranose, 3,4 6-tri-O-acetyl-2-deoxy-2-N-acetyl-beta-D-gluctosylamine and 2,3,6,2',3',4',6' hepta-O-acetyl-beta-D-lactosylamine as glycosyl donors, respectively. Target products (4, 7, 12a, 12b, 13) were obtained after deprotection. RESULTS: Five compounds (4, 7, 12a, 12b, 13) were synthesized as new compounds. The structures of all objective compounds were confirmed by 1HNMR, 13CNMR, HMQC, COSY, IR and MS. CONCLUSION: It was found that 12a showed better anti-inflammatory activity than (+/-) ibuprofen. PMID- 15127617 TI - [Isolation and structural identification of C-glycosylflavones from Corallodiscus flabellata]. AB - AIM: To study the chemical constituents of Corallodiscus flabellata. METHODS: Fresh plant of Corallodiscus flabellata was extracted twice with boiling water, concentrated to small volume under reduced pressure at 55 degrees C. The concentrated material was partitioned with ether, ethyl acetate, and n-butanol. The fraction of ethyl acetate extract was chromatographed over macroporous adsorption resin (Diaion HP-20) eluted with a mixture of H2O and MeOH in increasing MeOH content. Their fractions from resin were repeatedly chromatographed over Sephadex LH-20, silica gel column chromatography. The compounds were identified on the basis of their physiochemical and spectral data. RESULTS: Seven compounds were obtained and identified as 5,3',4'-trihydroxyl-6,7 dimethoxyl-8-C-[beta-D-xylocopyranosyl-(1-->2)]- beta-D-glucopyranosyl flavone (1), 5,4'-dihydroxyl-6,7-dimethoxyl-8-C- [beta-D-xylopyranosyl-(1-->2)]-beta-D glucopyranosyl flavone (2), 5,4'-dihydroxyl-6,7-dimethoxyl-8-C-beta-D glucopyranosyl flavone (3), 5,4'-dihydroxyl-6,7-dimethoxyl-8-C-[beta-D apiofuranosyl- (1-->2)]-beta-D-glucopyranosyl flavone (4), 3,4-dihydroxyl benzoyl acetic acid glycol ester (5), hydroxytyrosol (6), apocynin (7). CONCLUSION: Compound 1 and 2 are new compounds, the others were isolated from Corallodiscus flabellata for the first time. PMID- 15127618 TI - [Structural identification of huperzinine C]. AB - AIM: To study the alkaloid constituents of Huperzia serrata (Thunb.) Trev.. METHODS: Chromatographic methods were used for the isolation and purification. Structure was elucidated on the basis of chemical analysis and spectroscopic data. RESULTS: An alkaloid constituent was isolated from H. serrata (Thunb.) Trev.. CONCLUSION: The compound was found to be a novel lycodine type alkaloid with tricyclic structure named huperzinine C. PMID- 15127619 TI - Isolation and identification of a novel ellagitannin from Phyllanthus urinaria L. AB - AIM: To investigate the chemical constituents of Phyllanthus urinaria L. METHODS: Various chromatographic techniques were employed for the isolation and purification. The structure was elucidated by spectral analyses. RESULTS: A novel ellagitannin named phyllanthusiin G was isolated, its structure was established as 1-O-galloyl-2-phyllanthoyl-3,6-(R)-HHDP-beta-D-glucose. CONCLUSION: Phyllanthusiin G is a new compound. PMID- 15127620 TI - Determination of loratadine in human plasma by HPLC with fluorescence detector and study on its bioavailability. AB - AIM: To establish an HPLC-fluorescence method for determination of loratadine in human plasma and evaluate its relative bioavailability. METHODS: An Alltech-C18 column and a mobile phase of acetonitrile-water-glacial acetic acid-triethylamine (90:100:6:0.15) were used. The fluorescence detector was set at Ex 274 nm, Em 450 nm. The flow rate was 1 mL.min-1. RESULTS: The calibration curve was linear over a concentration range of 0.2-30 micrograms.L-1. The limit of quantification was 0.2 microgram.L-1. The average method recoveries varied from 96% to 98%. The results showed AUC, Tmax, Cmax and T1/2 beta between the testing tablets, testing capsules and reference tablets had no significant difference (P > 0.05). Relative bioavailabilities were 107% +/- 17% and 100% +/- 14% respectively. CONCLUSION: The three formulations were bioequivalent. PMID- 15127621 TI - [Identification of Panax notoginseng and its preparations by LC/MS]. AB - AIM: To develop a method of identifying the existing of Panax notoginseng in products of traditional Chinese medicine compound Danshen. METHODS: Total ion chromatograms (TIC) of Panax notoginseng, P. ginseng, and P. quinquefolius were obtained by means of LC/MS. Extracted ion chromatograms (EIC) posses m/z of 770, 800, 932, 946 and 1,108 of above-mentioned three herbs was compared between species. EIC 800 and 946 were selected as differentiation marks to distinguish P. notoginseng from the other two species. The EIC 800 and 946 of P. notoginseng were also compared to the EICs obtained by the same method from Chinese patent medicines of compound Danshen pellet, compound Danshen tablet, and compound Danshen injection. EIC 800 and 946 of P. notoginseng and its products possess similar peaks, relative retention time, and relative integral areas. Main chemical constitutes of P. notoginseng were also identified by using LC/MS/MS. RESULTS: EIC 800 and 946 were obviously different between P. notoginseng, P. ginseng, and P. quinquefolius. Patent medicines of compound Danshen pellet, compound Danshen tablet, which consist of extractions from P. notoginseng, possess the characteristic EICs. The selected EICs were stable and reproductive. CONCLUSION: EIC 800 and 946, which correspond to ginsenoside Rg1, Re, and their isomers, can be used as identifying mark of P. notoginseng to differentiate it from other herbs, and also can be used to tell apart P. notoginseng from other herb extractions in Chinese patent medicines of compound Danshen. PMID- 15127622 TI - [Determination of lornoxicam in human plasma by LC/MS/MS]. AB - AIM: To develop a sensitive and specific LC/MS/MS method for determination of lornoxicam in human plasma and investigate pharmacokinetics of single dose of lornoxicam in healthy Chinese volunteers. METHODS: Lornoxicam and the internal standard piroxicam were extracted from plasma using liquid-liquid extraction, then separated on a Zorbax XDB-C8 column. The mobile phase consisted of methanol water-formic acid (80:20:0.5) at a flow-rate of 0.7 mL.min-1. A Finnigan TSQ tandem mass spectrometer equipped with atmospheric pressure chemical ionization source was used as detector and operated in the positive ion mode. Selected reaction monitoring (SRM) using the precursor-->product ion combinations of m/z 372-->121 and m/z 332-->121 was used to quantify lornoxicam and internal standard, respectively. RESULTS: The linear calibration curves were obtained in the concentration range of 2.0-1,600 micrograms.L-1. The limit of quantitation was 2.0 micrograms.L-1. The method was successfully used in the pharmacokinetic study for lornoxicam. The main parameters obtained after an oral dose of 8 mg lornoxicam to 18 Chinese male volunteers were as follows: the value of T1/2 was (4.7 +/- 1.1) h, AUC0-infinity was found to be (5.5 +/- 2.4) mg.h.L-1. However, T1/2 of 105 h and AUC0-infinity of 189.5 mg.h.L-1 were obtained for another volunteer. CONCLUSION: The method is proved to be suitable for clinical investigation of lornoxicam pharmacokinetics, which offers advantages of specificity, speed, and higher sensitivity over the previously reported methods. PMID- 15127623 TI - [Pattern recognition applied to the fingerprint of traditional Chinese medicine characterized by two-dimension information data]. AB - AIM: To establish the two-dimension information fingerprints chromatograph of traditional Chinese medicine and investigate via pattern recognition. METHODS: Hierarchical cluster was applied to fingerprints described by one-dimension information data and by two-dimension information data respectively and corresponding results was compared. RESULTS: The gross classes, in which the samples denoted by two-dimension information data, classified by all kinds of hierarchical cluster methods had less differences and more robustness than the methods containing the samples expressed by one-dimension information data. The classes to which the unknown samples would be belonged were determined by hierarchical cluster analysis and artificial neural network (ANN), and the same results were obtained. CONCLUSION: The fingerprints characterized by two dimension information data could provide more overall and special information as compared with the methods indicated by one-dimension information data. PMID- 15127624 TI - [Effects of protease inhibitors on intestinal absorption of insulin]. AB - AIM: To study the effects of protease inhibitors on the large and small intestinal absorption of insulin in rats and to explore the mechanism of various protease inhibitors in different intestinal regions. METHODS: The intestinal absorption of insulin was evaluated by its hypoglycemic effect and serum insulin level using an in situ loop method with the washing treatment. RESULTS: Administration of insulin alone did not decrease the glucose level at either intestinal region with or without the washing treatment. With the unwashing treatment, there were no hypoglycemic effects in small intestinal loop when coadministration of insulin with protease inhibitors. With the washing treatment, the biological effects of insulin were amplified a little in small intestinal loop; obvious hypoglycemic effects were found in large intestinal loop with or without the washing treatment. The effectiveness of protease inhibitors was susceptible to their categories, concentrations and activities of proteolytic enzymes in different regions. The efficacy order of various protease inhibitors for enhancing hypoglycemic response of insulin was: leupeptin > sodium glycocholate > bacitracin > bestatin > cystatin; the percutaneous enhancement effects were observed in the presence of either sodium glycocholate or bacitracin. CONCLUSION: Coadministration of protease inhibitors could increase the insulin efficacy more effectively in the large intestine than in the small intestine. PMID- 15127625 TI - Effect of receptor solution specific conductivity on iontophoresis of tetracaine hydrochloride. AB - AIM: To examine the quantitative relationship between solution specific conductivity and the permeability of tetracaine HCl, and to investigate the effect of receptor solution specific conductivity on the iontophoretic transport. METHODS: An in vitro study was carried out to determine the iontophoretic permeability of tetracaine hydrochloride through rat skin. Iontophoretic flux of tetracaine hydrochloride through excised rat skin was determined using Valia Chien two-chamber diffusion cells with a constant d.c. current and Ag/AgCl electrodes. The specific conductivities of donor and receptor solution were also measured. RESULTS: Iontophoretic flux of tetracaine hydrochloride increased with a decrease of anion (chloride ion) concentration in receptor. And the iontophoretic permeability (ER, ER is the enhancement ratio, and ER = iontophoretic flux/passive flux) for tetracaine hydrochloride was directly related to the conductivity of receptor solution when other conditions were held constant. Linear regressions confirmed that ER was related to inverse of overall specific conductivity of donor and receptor solution [1/(ks.d + ks.r), ks.d and ks.r are the specific conductivity of donor and receptor solution]. CONCLUSION: The results suggest that specific conductivity of receptor solution may be a important factor for the iontophoretic permeability of a solute. PMID- 15127626 TI - [Studies on paclitaxel-loaded nanoparticles of amphiphilic block copolymer]. AB - AIM: To investigate the paclitaxel-loaded nanoparticles of poly(ethylene glycol) b-poly(D,L-lactic acid) amphiphilic diblock copolymer (PMT). METHODS: PMT was prepared by solid dispersion technique. The average size and size distribution were determined by dynamic light scattering (DLS). The morphology was characterized by transmission electron microscopy (TEM) and 1HNMR. The influences of the copolymer molecular weight and the paclitaxel-fed amount on PMT were studied. Therapeutic effect of PMT was studied on Kunming mice liver cancer H22. RESULTS: PMT showed nanometer size and spherical morphology with core and shell. The sizes of PMT increased with increasing the molecular weight of the hydrophobic segment in PEDLLA or increasing the drug-loaded amount. The tumour inhibiting effect of PMT was similar with that of Taxol. CONCLUSION: It will provide an experiment basis for the development of new kind of intravenous administration of paclitaxel. PMID- 15127627 TI - [Biotransformation of taxanes]. PMID- 15127628 TI - [Peroxisome poliferator-activated receptors and their modulators]. PMID- 15127629 TI - Can you make a merger pay off? AB - When three perinatal departments in three hospitals merged into a single department, the transition was complex, but cost-effective. PMID- 15127630 TI - Defeating malpractice risk, Part 1. AB - In this two-part series, learn the basis of malpractice, the areas where your nurses' risk is greatest, and how you can help safeguard your employees. Test your knowledge with the questions below, then check your answers at http://www.nursingmanagement.com. PMID- 15127631 TI - Best number, best mix, best method. AB - Learn the Joint Commission's requirements for staffing levels and skill mix and how surveyors determine if your organization measures up. PMID- 15127632 TI - Cruise to compliance. AB - Hospital leaders set a goal to achieve Accreditation with Commendation. In 3 years, they maximized resources, created a preparation process, and educated employees in a cruise-themed program to reach their goal. PMID- 15127633 TI - We made a mistake. AB - When Annals of Internal Medicine published a paper about the risk-management program at Veterans Affairs Medical Center, Lexington, Ky., readers were surprised at the facility's open policy regarding medication errors. Here's how health care leaders there put their liability on the line for better patient care. PMID- 15127636 TI - Listen for the "whispering disease". Learn the subtle signs and symptoms of ovarian cancer. PMID- 15127637 TI - Keeping in touch. Learn how to examine your breasts, then do it every month for a lifetime. PMID- 15127638 TI - Do I have breast cancer? PMID- 15127640 TI - Nutrition matters. PMID- 15127639 TI - Stop cervical cancer in its tracks. PMID- 15127641 TI - Conscious sedation ... coming to a unit near you. AB - Conscious sedation is becoming more prevalent for patients undergoing therapeutic and diagnostic procedures. Learn how to revise your conscious sedation guidelines to provide a consistent standard of care for all units--including clinical, environmental, and staff-related requirements. PMID- 15127642 TI - Infusion perfusion: i.v. pumps for every need. AB - Infusion pumps offer many options to improve patient care. Learn the latest technology and use this guide to choose the right pumps for your units. PMID- 15127643 TI - [Postoperative biliary stenosis: long-term results of endoscopic treatment]. AB - BACKGROUND: Endoscopic stent placement is often the initial therapy in symptomatic patients with postoperative strictures because patients are usually diagnosed at the time of ERCP. Although stent insertion rapidly relieves symptoms of biliary obstruction and can even be live-saving in patients with cholangitis, all stents eventually clog, necessitating regular stent changes every 3 to 4 months. Results from several groups suggested that placing multiple stents for months to years could dilate the stricture permanently and thus also treat patients with postoperative biliary strictures palliatively. OBJECTIVE: The outcome of temporary biliary stent placement for postoperative bile duct stenosis was retrospectively evaluated. This is a review of our experience with endoscopic dilation and stent placement in postoperative biliary strictures. PATIENTS AND METHODS: Thirty patients with postoperative strictures diagnosed with ERCP were treated with long-term endoscopic stent placement. One 10 Fr stent was placed at first whenever possible, and stents were exchanged every 3 months for a total of 18 months as median. RESULTS: Four men and 26 women with mean age 42 years (range 16-69 years), and laparoscopic cholecystectomy in six and cholecystectomy (open procedure) in 24; surgical history was reviewed retrospectively. Five patients were lost to follow-up and 25 patients were followed for a median of 18 months. In all 25 patients, previous to stent placement, 8.5 or 10 Fr, a mechanical or hydrostatic dilation was necessary. Stents were exchanged every 3 months to avoid cholangitis caused by clogging. Three 10 Fr stents were inserted in one patient, two 10 Fr stents in 14 patients, one 10 Fr and one 8.5 Fr stent in nine patients, and in one patient, one 10 Fr stent. Six patients (24%) developed recurrent stenosis and required surgery. CONCLUSIONS: Endoscopic treatment with mechanical or hydrostatic dilation and stent insertion may improve long-term results for patients with postoperative biliary strictures. PMID- 15127644 TI - [Hepatocellular carcinoma in youth: a comparative analysis with hepatocarcinoma in adults]. AB - INTRODUCTION: Hepatocellular carcinoma (HCC) is a very rare disease among young individuals. Epidemiological, clinical, and histopathological features of this malignancy in youth have not been thoroughly studied. PATIENTS AND METHODS: A review of clinical files of patients with HCC < 40 years of age treated between May 1990 and July 2002 was performed. RESULTS: Seventeen patients were included for analysis, nine were female and eight, male. Mean age at diagnosis was 24 years (range 12-39 years). Abdominal pain was the main symptom, followed by vomiting and nausea. Enlargement of liver was observed in 11 patients (65%). In seven patients (41%), etiologic factor was not found. Five of these cases were of fibrolamellar variant (29%). Only four patients were resected (23%) two of whom belonged to fibrolamellar type. Three patients (18%) are still alive after 64.9 months of follow-up. CONCLUSION: HCC is a very uncommon disease in youth and affects similarly both genders. It is discovered at advanced stage. Hepatitis B and C are uncommon etiologic factors. Frequency of fibrolamellar carcinoma is higher in this age group. Although resection is more feasible, overall survival rates remain low. PMID- 15127645 TI - [Is endoscopy necessary to initiate medical treatment in primary-level medical care patients with not investigated dyspepsia?]. AB - INTRODUCTION: Approximately 2 to 5% of primary care consultations are patients with dyspepsia. One of the most controversial points among primary-care physicians and gastroenterologists is the diagnostic-therapeutic approach of the uninvestigated dyspeptic patient. AIMS AND METHODS: Our aim was to investigate prevalence of organic disease and Helicobacter pylori (Hp) status in uninvestigated dyspeptic out-patients at a primary-care level. Two hundred two adult out-patients (130 F/72 M), > or = 18 and < 75 years, mean age 36 years (range 18-73 years), were recruited and sent for upper gastrointestinal endoscopy and gastric biopsies for CLO-test due to dyspeptic symptoms according to Rome II criteria. Patients were excluded if they had previously been investigated by any means, had a background of NSAIDs or corticoesteroids intake, irritable bowel syndrome (IBS) or any alarm sign or symptom. All endoscopic findings were reported to primary care physicians who decided on patient management. RESULTS: A total of 86.14% (174/202 patients) showed no endoscopic lesions and only 13.86% (28/202) [95% CI = 9.1-18.6] showed evidence of non-malignant organic lesions. By age group, prevalence of organic diseases was 10.81% (16/148) [95% CI = 6.3-16.9] in patients < 45 years and 22.22% (12/54) [95% CI = 12.04-35.6] in patients > 45 years, OR = 0.424. [95% CI = 0.186-0.968]. CONCLUSIONS: Prevalence of upper gastrointestinal tract organic disease in dyspeptic out-patients selected according to Rome II criterion at a primary-care level is low, with no malignancies detected. These findings show that patients < 45 years of age with uninvestigated dyspepsia may empirically and safely receive symptom-guided antisecretory therapy. PMID- 15127647 TI - [Non-erosive gastroesophageal reflux disease. Incidence of clinical forms, characteristics of esophageal exposure to acid and symptomatic correlations]. AB - BACKGROUND: It is estimated that up to 70% of community-based patients with heartburn have no evidence of esophageal mucosal injury or non-erosive gastroesophageal reflux disease (NERD). Studies in NERD patients using esophageal pH monitoring and symptom index (SI) have suggested different subgroups or clinical forms. OBJECTIVE: Our aim was to investigate frequency of different subgroups of patients with NERD according to esophageal pH-monitoring parameters and SI, and to determine esophageal acid exposure characteristics and the relationship between symptoms and acid reflux in NERD. MATERIAL AND METHODS: Patients with heartburn at least 3 times per week during 12 weeks during the last year and negative upper GI endoscopy were studied. All patients were submitted to 24 h esophageal pH-metry and SI was calculated. Patients were classified into three groups: group I, patients with abnormal pH-metry; group II, patients with normal pH-metry and positive SI (> or = 50%), and group III, patients with normal pH-metry and negative SI (< 50%). Total number of reflux episodes, length of episode, % time pH < 4, and Johnson-DeMeester score were evaluated. RESULTS: Three hundred patients, 206 women and 94 men, were included. A total of 137 (46%) were classified in group I, 62 (21%) in group II, and 101 (33%) in group III. There were no significant differences in gender and age among groups. All pH parameters were significantly higher in group I. In group II, there was good correlation between heartburn and acid reflux. Esophageal acid exposure in group III was minimal and there was no relationship among symptoms and acid reflux episodes. CONCLUSIONS: Subjects with NERD represent a heterogeneous group of patients. Three subgroups of patients with NERD were identified by pH-metry and SI; a) patients with abnormal acid exposure; b) patients with hypersensitive esophagus (heartburn with physiologic amounts of acid reflux), and c) patients with no correlation between symptoms and acid reflux. PMID- 15127646 TI - [Role of polymorphism of certain cytokines in gastric cancer in Mexico. Preliminary results]. AB - BACKGROUND: Interleukin-10, tumor necrosis factor alpha, Interleukin-1 beta and interleukin-1 receptor antagonist cytokines modulate the inflammatory response in presence of Helicobacter pylori. Pro-inflammatory interleukin 10 (IL-10-592, 1082), TNF alpha (TNF alpha-308), interleukin-1 beta and interleukin-1 receptor antagonist (IL-1B-31*C and IL-1RN*2/*2) genotypes have been associated with higher risk of gastric cancer in Caucasians. The aim of this study was to investigate whether these same genotypes are involved in susceptibility to gastric cancer in Mexican population. MATERIALS AND METHODS: DNA from 33 unrelated Mexican patients with histologically confirmed gastric cancer (n = 25) or high-grade dysplasia (n = 8) (mean age 62.7, F/M = 0.37) and 25 ethnically matched healthy controls (mean age = 39.9, F/M = 3.12) were studied. All cases and controls had evidence of H. pylori infection as shown by at least two positive results from the following diagnostic tests: rapid urease test; culture; histology, or detection of IgG anti-H. pylori antibodies. The -592, -1082 polymorphism in IL-10 gene, the -308 in TNF alpha gene, and the-31 polymorphism in the IL-1B gene were typed by 5' nuclease PCR assays (TaqMan) and the variable number of tandem repeats polymorphism in intron 2 of the 1L-1RN gene was typed by PCR and amplicon sizing as previously described (Nature 2000; 404: 398). RESULTS: Carriage of the pro-inflammatory IL-1B-31*C allele was associated with increased risk of gastric cancer or high-grade dysplasia (OR: 8.7, 95% confidence interval [CI] = 1.5-66.9). No association was found between any IL-IRN, IL-10 or TNF alpha genotypes and gastric cancer or high-grade dysplasia. Logistic regression analysis identified male gender and carriage of IL-1B-31*C as independent risk factors for gastric cancer (OR = 9.2, 95% CI = 2.4-34.5, and OR = 10, 95% CI = 1.6-64, respectively). CONCLUSIONS: The results of this preliminary study confirm that the pro-inflammatory IL-1B genotypes, as well as male gender, are risk factors for development of gastric cancer in Mexican population. PMID- 15127649 TI - [Arterial hypertension as clinical manifestation of non-parasitic hepatic cyst. A case report]. AB - We present the case of a 32-year-old female patient referred to the Instituto Nacional de Cancerologia (INCan) with a history of several months of systemic hypertension and epigastric pain associated to early postprandial satiety, fatigue, and dyspnea. At physical examination, a smooth, non-tender, palpable mass was found in right upper quadrant 7 cm below costal margin. Computed tomography (CT) scan showed right-sided non-parasitic liver cyst. The patient was taken to the operating room, where she presented hypertensive crisis of 180/125 mm Hg, which did not respond to midazolam therapy. The procedure was suspended and deferred. After blood pressure control with angiotensin II-antagonist and calcium-antagonist, the patient was submitted to laparotomy, where a wide deroofing of the lesion was performed. The postoperative evolution was uneventful and the patient has remained normotense without anti-hypertensive medication after 6 months of follow-up. PMID- 15127648 TI - [Variations in a 24-year period of colorectal and gastric cancer in Mexico]. AB - OBJECTIVE: Gastric cancer (CG) and colorectal cancer (CCR) are the two most common neoplasms of the digestive system in the world. We performed a study to determine incidence and relation between CG and CCR in five hospitals in Mexico City. METHODS: Patients with admitted diagnosis of CG and CCR at Hospital General de Mexico, Instituto Nacional de Ciencias Medicas y Nutricion Salvador Zubiran, Hospital Espanol de Mexico, Centro Medico Nacional "20 de Noviembre" from the Instituto de Salud y Seguridad Social para Trabajadores del Estado, and Hospital Central Militar from January 1978 to December 2001 were studied. RESULTS: A total of 7,136 patients were studied. (CG 3,830, CCR 3,306). At Instituto Nacional de Ciencias Medicas y Nutricion "Salvador Zubiran" CG was the most common digestive neoplasm; from 1999, ratio was inverted to < 1. At Hospital General de Mexico, from the beginning and until 1984, ratio was > 2, and later had an average of 1.31. For Hospital Espanol, ratio always was < 1 without changes. At Centro Medico Nacional "20 de Noviembre", initially CCR was more frequent, then CG, and finally CCR. At Hospital Central Militar ratio was constant, > CG. At the beginning, was global behavior > CG, ratio seemed to invert, but since 1998 CG/CCR ratio was < 1 and continued that way. CONCLUSIONS: In this study, we found that changes of CG/CCR ratio in a period of 24 years showed elevation of CCR incidence at five Mexican hospitals. PMID- 15127650 TI - [Neuroendocrine gastric carcinoma. A case report and review of current management]. AB - Neuroendocrine or carcinoid tumors of the gastrointestinal tract considered previously extremely rare, are diagnosed at present with increased frequency due to the better capacity to identify neuroendocrine system cells in normal and pathologic conditions. Occasionally, these tumors secrete a great variety of vasoactive substances, producing the carcinoid syndrome. Gastric carcinoids are classified, according to their degree of differentiation into well differentiated and poorly differentiated tumors, also called neuroendocrine carcinomas. Neuroendocrine gastric carcinomas or poorly differentiated gastric carcinoids are seen in 5-15% of all gastric carcinoids, mainly in older male patients. Generally they are large, very aggressive tumors with extensive local infiltration. Due to poor differentiation, they are not frequently associated with an endocrine syndrome. They can be located in any part of the stomach but are mainly seen in antrum. These tumors have an aggressive behavior and must be treated in a radical manner; recurrences are not uncommon. We report the case of a patient with a neuroendocrine gastric carcinoma treated with an en bloc subtotal gastrectomy and colectomy. PMID- 15127651 TI - [Clinical images in gastroenterology. Appendicitis caused by a foreign body]. PMID- 15127653 TI - [Surgical treatment of peptic ulcer]. AB - Despite a decreasing number of operations for ulcer, there are many patients who require definitive treatment. If an operation is required for duodenal ulcer, vagotomy of some type is part of the treatment, and in gastric ulcer resection with or without vagotomy is required. Extended proximal gastric vagotomy can be performed in the majority of patients, excluding those who are unstable or have severe concomitant diseases. In cases of urgent surgery for hemorrhage or perforation, the surgical procedure must be selected individually. Although the role of traditional operations is well established, there is increasing interest in laparoscopic approaches. However, because there is a diminishing of elective surgery for ulcer, it is unlikely that these new procedures may be evaluated as operations were evaluated in the past. PMID- 15127652 TI - [Fascioliasis: diagnosis, epidemiology and treatment]. AB - Fascioliasis is a trematode, disease of liver and bile ducts of sheep, cattle, and other ruminants throughout the world that is caused by the fluke, Fasciola hepatica. Human infection has been reported in Mexico, Cuba, Puerto Rico, Chile, Peru, Uruguay, Brazil, Argentina, the US, Europe, eastern Africa, Japan and Australia. The parasite's miracidium invades one of the various Lymnaea water snail hosts. Infection results from ingestion of encysted metacercariae attached to raw watercress (Nasturtium officinale). Symptoms recorded from human cases included irregular fever, epigastric pain and abdominal tenderness, obstructive jaundice and leucocytosis with eosinophilea up to 60%. Specific diagnosis is based on recovery of the eggs in the patient's stool or from biliary tract drainage. Treatment is with emetine hydrochloride given intramusculary. Bithionol is given orally at a dosage of 30-50 mg/kg but on alternate days from 10 to 15 doses. Praziquantel is probably effective. Preventive measures include education of the public on mode of transmission of life cycle of the parasite, and dipping fresh watercress into boiling water for a few sec, or drying suspected watercress. PMID- 15127654 TI - [Variations in a 24-year period of colorectal and gastric cancer in Mexico]. PMID- 15127655 TI - Body building. PMID- 15127656 TI - Burning down to rock. PMID- 15127658 TI - Splash of cold water. PMID- 15127657 TI - Downsized target. PMID- 15127659 TI - Blue-collars in eclipse. PMID- 15127660 TI - Making drugs, not profits. PMID- 15127661 TI - Patents on ice. PMID- 15127662 TI - The myth of the beginning of time. PMID- 15127663 TI - Questions about a hydrogen economy. PMID- 15127664 TI - Synthetic life. PMID- 15127665 TI - Freud returns. PMID- 15127667 TI - Retooling the Global Positioning System. PMID- 15127666 TI - Freud returns? Like a bad dream. PMID- 15127668 TI - The transit of venus. PMID- 15127669 TI - Laser eye surgery. Clear favorite. PMID- 15127670 TI - Who Operates When? II: The National Confidential Enquiry into Perioperative Deaths 2003. PMID- 15127671 TI - Intermediate care and diversity: principles and practice. PMID- 15127672 TI - Evidence-based management of schizophrenia. AB - Antipsychotic medication is the foundation of care for patients who suffer from schizophrenia and other psychotic conditions. Newly diagnosed patients should be treated with atypical antipsychotics, of which the National Institute for Clinical Excellence guidelines recommend five. The quality of clinical evidence will ultimately establish the differences between available treatments. PMID- 15127673 TI - Long-term management of atherothrombosis. AB - While clinical trials of specific antithrombotic agents can reduce the risk of vascular events in at-risk patients, understanding the results of these studies is key to optimizing benefit and minimizing risk. PMID- 15127674 TI - Comorbid chronic non-cancer pain and opioid use disorders. AB - Patients with chronic non-cancer pain and opioid dependence are difficult to treat effectively. This article reviews the common issues that arise in relation to assessment and treatment, and recommends the adoption of an integrated approach to this patient population. PMID- 15127675 TI - Dopamine agonist switching in Parkinson's disease. AB - There are a number of situations for patients with Parkinson's disease in which a safe and efficacious switch from treatment with one dopamine agonist to another may be required. This article explores reasons for making such a switch, and provides practical guidance on performing it. PMID- 15127676 TI - Necrotizing enterocolitis. AB - Necrotizing enterocolitis is the principal cause of surgical referral in preterm neonates and the most common gastrointestinal emergency among infants. Its pathophysiology is unclear and it carries high levels of mortality and morbidity. This article provides an overview of necrotizing enterocolitis including risk factors, preventative strategies and medical and surgical management. PMID- 15127677 TI - Integrated cardiac care for improved pre-hospital thrombolysis. PMID- 15127679 TI - Diabetic emergencies in children. PMID- 15127678 TI - Orthopaedic manifestations of tuberculosis. AB - An estimated 1 million people will be infected with tuberculosis worldwide in the first 20 years of the 21st century. If the disease is not recognized and treated early the morbidity and mortality of this condition will rise. This article highlights the varying manifestations of tuberculosis from an orthopaedic perspective. PMID- 15127680 TI - Early signs of the trainee in difficulty. AB - A series of high-profile medical scandals and tragedies has generated increasing interest in the earlier detection of doctors whose health, conduct or performance may pose a risk to themselves or others. The earliest signs of problems often emerge during the training years, when it may be possible to take remedial action to prevent the doctor derailing. This article describes the early signs that have identified doctors in difficulty in a large postgraduate deanery. PMID- 15127681 TI - Parasites and the eye. PMID- 15127682 TI - Ibuprofen-induced eosinophilic pneumonia. PMID- 15127683 TI - Pulmonary hypertension: a rare but serious complication of ventriculoatrial shunts. PMID- 15127684 TI - Deep cholestatic jaundice and pulmonary hypertension in a woman with Graves' hyperthyroidism. PMID- 15127686 TI - Aggressive angiomyxoma of the vulva. PMID- 15127685 TI - Vaccination metastasis following percutaneous endoscopic gastrostomy. PMID- 15127687 TI - Images in medicine. Cystic duct stump stone. PMID- 15127688 TI - Use of the laryngeal mask airway in the prone position. PMID- 15127689 TI - Sailorproofing quality. PMID- 15127690 TI - Application of variables control charts to risk-adjusted time-ordered healthcare data. AB - In a previous article (M. K. Hart, Qual Manag Health Care. 2003;12(1):5-19), the authors presented risk-adjusted control charts applicable for attributes data. The present article discusses a similar class of control charts applicable for variables data that are often skewed. The key feature of these charts is their application of risk-adjusted data in addition to actual performance data. The resulting charts should decrease the occurrence of both type I and type II errors as compared to the unadjusted control charts. This article presents several control charts that vary in the data transformation and combination approaches. Data depicting hospital length of stay following coronary artery bypass graft procedures were used to illustrate the use of transformed and risk-adjusted control charts. PMID- 15127691 TI - SARS: a quality management test of our public health safety net. AB - Emergence of severe acute respiratory syndrome in March 2003 tested all aspects of BC Centre for Disease Control (BCCDC) operations. In addition to its public health responsibilities, BCCDC was pivotal in the science defining SARS. These events occurred under international scientific and media scrutiny over a 4-month period and were seen as an opportunity to learn about how the Centre performed under extreme pressure as a QM-based (quality-management-based) organization. A retrospective review of the QM practices over the previous 6-months was initiated on June 30, 2003. Key management documentation during the study period was reviewed. Structured interviews were conducted with front line personnel. Customized instrumentation was developed to correlate management decisions with recognized QM criteria: anticipatory management; keeping programs on track; ongoing adjustment, improvement, and revision; identifying and improving sources of error, waste, and redundancy; feedback from key stakeholders; and data-driven decision-making methods. The team structure between laboratory science and epidemiology was critical. This was attributed to the culture of scientific discovery of the organization. All knowledge gained was shared with other organizations around the world. The consensus is that British Columbia was very lucky this time around. This review is part of BCCDC's commitment to fighting emerging infectious diseases. PMID- 15127692 TI - Getting going together: can clinical teams and managers collaborate to identify problems and initiate improvement? AB - A clear aim is key for the success of improvement projects, yet many fail already at this stage. We studied how clinical teams and managers at a university hospital in Sweden identified problems and defined aims as they initiated 24 process improvement projects. Categorizing and comparing problems at 3 stages of problem definition, we found that the majority of problems fell into 1 of 3 categories: information issues, poor procedures, and waiting times. Going through these stages, managers and clinical teams prioritized waiting-time problems. We show how managers can ask such teams to quickly identify problems suited for improvement projects through this step-wise, facts-based approach. We conclude that they can add their management perspective when giving specific assignments, to harness the combined benefits of both a bottom-up and a top-down approach to improvement. PMID- 15127693 TI - Is time-slice analysis superior to total hospital length of stay in demonstrating the effectiveness of a month-long intensive effort on a medicine service? AB - To control the upward spiral of healthcare costs, hospitals seek to implement efficiency interventions whose benefits are frequently assessed by reductions in average inpatient length of stay (LOS). However, average hospital LOS is a crude metric when trying to assess the utility of an intervention focussed on a particular service or over a specific time window. It cannot isolate the time or place of the intervention from the full duration of a patient's hospital visit, which may include more than 1 hospital service or extend beyond the intervention's time window. At Montefiore Medical Center, a new analytic method was used to describe a month-long effort to improve care efficiency in a hospital teaching service. Using an extension of the Cox proportional hazard model (S plus), we were able to analyze the contribution of only those patient-days that took place during the time window of interest on the service of interest, eliminating the contamination of the "non intervention days." Having built the appropriate model, we were then able to graph the behavior of the groups with and without the intervention and calculate the model's expected average LOS, controlling for the appropriate variables. By comparing this method with a conventional average LOS analysis, we demonstrate the superiority of using this "time slice" method over the conventional analysis of LOS. PMID- 15127694 TI - [Aortic stenosis: interest in echocardiography with dobutamine]. AB - The stress echocardiography with dobutamine confirms her increasing value for the patients with severe aortic stenosis and left ventricular dysfunction in a diagnosis concept by selecting the true severe stenoses and revealing stenoses overestimated by the low fow, and especially prognosis by means of the stratification of the operatory risk and the search for a contractile reserve which represent the guarantee of a survival per and postoperatoire satisfactory. As regards asymptomatic aortic stenosis the times of intervention are in reevaluation, the compliance valvulaire represent reliable means which needs confirmation for a prophylactic indication for surgery. PMID- 15127695 TI - [Biventricular resynchronization in the management of severe cardiac insufficiency]. AB - Heart failure is a major problem of public health, it represents a frequent status among patients with heart disease, and its implications in term of mortality and cost are high. Non Pharmacological treatment of heart failure most commonly designed as cardiac resynchronization therapy (CRT) has demonstrate efficacy to improve functional class, exertion capacity, left ventricular ejection fraction, reduction of mitral regurgitation, and probably mortality at midterm. The most recent studies emphasize on the role of implantable cardioverter defibrillate or (ICD) combined with CRT to reduce mortality. More trials are needed to valid this concept. PMID- 15127696 TI - [Long-term outcome of surgically treated teratology of Fallot]. AB - Tetralogy of Fallot is the most common cyanotic congenital heart disease. The surgical treatment that is palliative or complete repair has allowed to transform the preview of this heart disorder. We suggests to study the long term outcome in patients undergoing surgical repair of tetralogy of Fallot, by emphasizing the quality of their lives, the complications, as well as the mortality. Ventricular arrhythmia and sudden cardiac death after repair of tetralogy of Fallot are devastating complications in adults survivors and their prediction remains difficult. PMID- 15127697 TI - [Anatomic repair of transposition of the great arteries or arterial switch operation. Report of 62 cases]. AB - Between January 1990 and September 2003, 62 patients underwent anatomic repair of a transposition of the great arteries. Mean operative age is 40 days. Transposition of the great arteries was simple in 38 cases and associated to a large ventricular septal defect in 24 cases. 44 patients have had an atrial septostomy of Rashkind and 45 an infusion of prostaglandin E 1.5 patients with simple transposition of the great arteries have had left ventricular retraining before arteriel switch. In association to arterial switch, were performed closure of ventricular septal defect in 24 cases, cure of coarctation of the aorta in 4 cases and cure of an abnormal partial pulmonary venous return in 1 case. Early mortality was 6,45%. After a mean follow up of 3 years, one patient died suddenly (late mortality is 1.72%) and one patient had to have 2 reoperations. Results of anatomic repair are now excellent. Late mortality is essentially related to coronary complications so that a careful follow-up is mandatory. PMID- 15127698 TI - [Left coronaroventricular microfistula]. AB - Left coronaroventricular microfistulae is a rare malformation. The authors report 11 cases of microfistulae between coronary arteries and left ventricle diagnosed by coronary arteriographies. These cases include 6 men and 5 women. Patient's mean age was 54.4 years. The symptoms were suggestive of coronary pathology. The ECG showed myocardial ischemia signs in 5 cases. The other patients had a positive exercise-test. Microfistulae originated from the left anterior descending artery were seen in 5 cases, from the right coronary artery in 2 cases, from the circumflex in 1 case and from the lateral artery in 1 case. The microfistulae originated from both left anterior descending artery and right coronary artery were observed in 2 patients. The main mechanism of myocardial ischemia seems to be related to the coronary steal phenomenon. The diagnosis of the microfistulae is based on coronary arteriography with late recorder angiographic images. The treatment is essentially medical. Surgical and transcatheter treatments are exceptional and must be considered in only severe forms with refractory medical treatment. PMID- 15127699 TI - [Congenital coronary aneurysm. Three case reports]. AB - Congenital coronary aneurysms are an unusual anatomical entity. Their prognosis appears to be particularly dependent on the presence or absence of aneurysm thrombosis. We report three cases of congenital coronary aneurysms, diagnosed in one case after myocardial infarction. Two patients were treated successfully by an exclusion of the aneurysm and coronary bypass and the third patient was treated medically. The aim of this study is to discuss the clinical features, prognosis and management of this disease. PMID- 15127700 TI - [Percutaneous mitral commissurotomy and left ventricular function in severe mitral stenosis]. AB - The purpose of this study was to determine the immediate effect of percutaneous mitral commissurotomy (PMC) on left ventricular (LV) performance. We studied 30 patients with severe mitral stenosis undergoing successful PMC by cardiac catheterization and angiography before and 5 minutes after PMC. All patients were in sinus rhythm and no patient had hypertension or clinical coronary artery disease. We conclude that the left ventricular end-diastolic volume and the stroke volume increased significantly immediately after PMC because of and increase in LV filling after relieving the mitral mechanical obstruction. PMID- 15127701 TI - [Results of coronary angioplasty in coronary bifurcation. Report of 61 cases]. AB - These are the results of 61 bifurcations treated by percutaneous coronary angioplasty in 50 patients (41 males, 9 females) between 1998 and 2003. Bifurcation stenosis, dominated by type I of bifurcation classification. Global restenosis rate was 20% and didn't concern any case of kissing balloon. Restenosis rate in bifunction angioplasty is similar to that of the other sites; besides, it's twice more important when we stent both of the principal and collateral arteries unless we did kissing balloon. Te interventional treatment of bifurcation stenosis is feasible, with restenosis rate similar to the other types of lesions if we proceed systematicaly to kissing balloon. PMID- 15127702 TI - [Radiofrequency ablation of accessory atrioventricular pathways. Predictive factors of immediate and long-term results]. AB - The purpose of this study was to evaluate safety and efficacy of catheter ablation of accessory pathways (APs) and to identify predictive factors of acute result and outcome. The patient population included 173 patients who had undergone ablation of an AP. The success rate was 91.6%, a major complication occurred in 4 patients (2.3%) and 8 patients (4.9%) developed a recurrence after a successful ablation procedure. Only one factor predicted success (sex), tow variables predicted development of a major complication (septal AP, age < 16 years), and three factors predicted arrhythmia recurrence (mid septal, right free wall, and multiple APs). CONCLUSION: The results of this study may serve to identify sub-groups of patients most likely to have a favourable result in whom it would be reasonable for clinicians to recommend catheter ablation as first line therapy. PMID- 15127703 TI - [Therapeutic strategies of atrial fibrillation in patients aged 65 and over. Report of 86 cases]. AB - Auricular fibrillation is the most common sustained cardiac arrhythmia, yet the optimal strategy for its management remains unclear. Since the study AFFIRM from which resulted them final were deferred in December 2002, the assumption of responsibility of the fibrillation of the old subject tends to become clarified. 86 patients at least 65 years or more were enrolled in our retrospective study from January 1997 to June 2003. The mean age is 70 years (65 to 82 years) and sex ratio to 0.79. 34% had a history of hypertension and 12% had a coronary artery disease. 2 groups were individualized, according to the year of admission before or after 2002. It comes out from these work 2 points: the restoration of the sinusal rhythm was the first choice. In the event of failure, the option was to maintain fibrillation and this, in the 2 groups. The AVK were founded in the large majority of the cases only after year 2002. PMID- 15127704 TI - [Role of fibrin D-dimer in acute coronary syndrome. Prospective study of 22 cases]. AB - Fibrin D-dimer are the consequence of an excess of fibrinolysis. The raise of their level in coronary heart disease seems to be helpful to enhance the diagnosis of coronary ischemia. Prospective study over 4 months, including 22 patients (16 male, 6 female) divided in 2 subgroups: Group I: 10 patients investigated for stable angina Group II: 12 patients investigated for ACS without ST elevation. All patients underwent fibrin D-dimer dosage and coronarography. Fibrin D-dimer levels were higher in group II (924.5 ng/ml vs 703.9 ng/ml; p < 0.0001). In group II, 6 patients had ST depression with a level of fibrin D-dimer 879.5 ng/ml vs 969.6 ng/ml in the other 6 patients. We found a positive correlation between level of fibrin D-dimer and complexity of coronary lesions (1007 ng/ml in type C vs 675 ng/ml in type A lesions; p < 0.0001). Fibrin D-dimer seems highly implicated in coronary disease and if these results are confirmed by larger studies their routine dosage will be helpful in ACS. PMID- 15127705 TI - [Aortic infectious endocarditis on native valve]. AB - Between 1990 and 2002, 32 patients with mean age of 34 years has been treated for an aortic infective endocarditis on native valve. All the patients had recognised heart disease before developing the infection. 24 patients are operated for aortic valve replacement. The hospital mortality was 15.6% (5 patients). The mean follow-up was 47.4 months. The survival rate was 75%. Aortic infective endocarditis remains a serious affection with poor prognosis, despite the contribution of echocardiography and advances in antimicrobial therapy and cardiac surgery. Significants morbidity and mortality are usually caused by sequele of the disease rather than by the infection itself. PMID- 15127706 TI - [Surgically treated chronic aortic insufficiency: prognostic value of preoperative echocardiography]. AB - Between March 1979 and December 1998, 38 patients with isolated chronic aortic insufficiency underwent aortic valve replacement. The aim of this study is to determine whether preoperative echocardiography parameters are useful in predicting operative results in patients with aortic valve replacement for chronic aortic insufficiency. The global survival is 92% in one year and 89% in ten years. We conclude that on end systolic left ventricle diameter < 55 mm has good prognosis (p = 0.019) and there is a strong correlation between preoperative end systolic and post operative end systolic diameter an end diastolic left ventricular diameter > 70 mm and a left ventricular fractional shortening < 25% weren't predictive of a poor prognosis. PMID- 15127707 TI - [Cardio-pericardial hydatid cyst. Report of 19 cases]. AB - From May 1991 to July 2003, 19 patients (9 men and 10 women), of 30 years average age were operated in our department service for surgical cure of cardio pericardial hydatid cysts. The TEE constitutes the examination of choice in the cardio-pericardial diagnosis of hydatid cyst. The surgery under CPB is the technique of choice, we practiced used 15 times. The left ventricular localization is most frequent. We deplore only death into post-operative immediate. The post-operative was course simple at the majority of our patients. 17 patients could be controlled remotely of the intervention with 35.5 months an average passing. We deplore 2 late deaths. No relapse was observed. PMID- 15127708 TI - [Predictive factors of mortality in combined coronary and vascular surgery. Report of 46 cases]. AB - From 1994 and 2003, 46 patients underwent surgical myocardial revascularisation associated to valvular surgery. Surgical indications were for valvular disease in 13 cases (28%), for coronary disease in 16 cases (35%) and associated coronary and valvular disease in 17 cases (37%); Isolated conservative mitral repair was performed in 22 cases (48%); mitral valve replacement in 10 cases (41%), aortic valve replacement in 11 cases (46%) associated mitral and aortic surgery in 3 cases (13%). Intra-aortic balloon pump was used in 4 patients (9%). In the post operative period complete atrio-ventricular heart bloc occurred in 2 cases. For the entire group, 8 deaths (17%) occurred in the hospital, related to low cardiac output in 5 cases (63%). Long term follow up was achieved in a mean period of 18 months. 25 patients (68%) were asymptomatic and one late death occurred after severe heart failure. The predictive factors of hospital mortality were: preoperative ischemic mitral insufficiency. Various large series in the literature have documented operative risk of myocardial revascularisation when combined with valve surgery. In our group the high rate of hospital mortality (17%) explains the difficulties in medical and surgical care of this combined valvular and coronary disease especially in the perioperative period. PMID- 15127709 TI - [Correlation of clinical and angiographic morphology in unstable angina]. AB - The goal of this study was to compare the clinical presentation and angiographic morphology of patients having an unstable angina pectoris. A total of 321 patients were consecutively studied and underwent cardiac catheterization, mean age 59 + 6 years. According to Braunwald classification, class III was predominant (58%) On coronary angiography, 148 patients had single vessel disease, double-vessel in 92 and triple-vessel in 64. Morphology of coronary artery lesions was classified according to Ambrose's classification, 100 patients had simple lesions (type I or IIA), 204 patients had complex lesions (type IIB, III, intracoronary thrombus or total occlusion). Thoracic rest pain (class III) or postinfarction angina (class C), were associated with the presence of complex lesions. This subgroup of high risk patients would benefit from either Glycoprotein IIb/IIIa blockers with an early revascularisation strategy. PMID- 15127711 TI - [Pulmonary hypertension in sickle cell anemia. A case report]. AB - We report the case of a 33-years-girl with SCA and severe PH. She developed six month before admission, non productive cough and dyspnea. Physical examination at admission revealed shortness of breath and right heart ventricular failure. Electrocardiography showed sinus rhythm and an incomplete left bundle branch block. Chest roentgenography revealed cardiomegaly with cardiothoracic index at 0.66 and pulmonary infiltrates. Laboratory tests revealed an anemia with hemoglobin of 7.1 g/dl, white blood cell count of 12,500/mm, moderate renal failure (cretininemia = 178 mumol/l) and hypoxemia with oxygen pressure of 60 mmHg. Hemoglobin electrophoresis revealed on heterozygous SCA. Echocardiography revealed dilatation of right heart cavities and a systolic pulmonary artery pressure of 60 mmHg. A perfusion lung scintigraphy demonstrated multiple subsegmental perfusion defects. PH is a common complication of adult patients with SCA. Appropriate therapies and strategies for prevention of PH in SCA are unknown. Further research exploring therapies such as oxygen, nitric oxide, prostacyclin and hydroxyurea are indicated. PMID- 15127710 TI - [Myocardial infarction revealed by ischemic stroke]. AB - Ischemic cerebral infarction associated with myocardial infarction is yet a real diagnosis challenge. If during the acute myocardial phase the mechanism is mostly embolic, at long-term, the mechanism is not clearant and other causes should be searched. We report a 50 year old man with ischaemic stroke with strong evidence of myocardial infarction in the late phase with wall-motion abnormality and mural clot revealed by echocardiography and Q waves. Atrial fibrillation was suspected and no other abnormalities could be found. The diagnosis of cardio-embolic ischaemic stroke could not be made with certainly. PMID- 15127712 TI - Does an echinacea preparation prevent colds? The debate continues. PMID- 15127713 TI - Changing labels: lice treatments...and changing ingredients: Kaopectate. PMID- 15127714 TI - Does breastfeeding prevent obesity?...and what about dairy foods? PMID- 15127715 TI - Not all child safety seats are equal. PMID- 15127716 TI - Pivoting on p53. PMID- 15127717 TI - Unfolding targets for dengue fever. PMID- 15127718 TI - Delivering on immunosuppression. PMID- 15127719 TI - Primary hyperoxaluria type 1: appearance in FDG PET, bone- and bone marrow scintigraphy. PMID- 15127720 TI - Incidental diagnosis of asymptomatic pericardial effusion in uraemic patient by using 99mTc-MIBI perfusion SPECT. PMID- 15127721 TI - [ESPEN guidelines for evaluating nutritional status 2002]. PMID- 15127723 TI - Cell adhesion in mammary gland biology and neoplasia. PMID- 15127724 TI - Von Willebrand's disease: a case study. PMID- 15127722 TI - Pancreatic carcinoma with positive iodine uptake lacking NIS-protein-expression. PMID- 15127725 TI - Versatile pharmacological actions of YC-1: anti-platelet to anticancer. AB - Since the first article on YC-1 was published in 1994, it has been popularly used as a pharmacological tool to activate soluble guanylate cyclase and to increase cyclic GMP levels in cultured cells or isolated tissues. In terms of the pharmacological actions of YC-1, previous studies tend to be limited to it inhibition of platelet aggregation and vascular concentration. However, recent studies have demonstrated that YC-1 has versatile pharmacological effects other than the anti-platelet and vasodilatory effects. In particular, two recent reports suggest that YC-1 could be developed as a new class of anticancer agent for rapidly growing solid tumors, because it inhibits hypoxia-inducible factor 1 (HIF-1) activity, and has been reported to halt tumor growth in vivo. We here review the cyclic GMP-dependent and independent pharmacological actions of YC-1, and its anti-HIF-1, anticancer effect. PMID- 15127727 TI - A novel technique to reduce curved needlestick injuries. PMID- 15127728 TI - The use of corrugated drains in the management of the infected diabetic foot. PMID- 15127726 TI - Correlation of DNA adduct formation and riddelliine-induced liver tumorigenesis in F344 rats and B6C3F1 mice [Cancer Lett. 193 (2003) 119-125. AB - Riddelliine is a naturally occurring pyrrolizidine alkaloid that induces liver hemangiosarcomas in male and female F344 rats and male B6C3F1 mice. We previously reported that eight dehydroretronecine (DHR)-derived DNA adducts were formed in liver DNA of rats treated with riddelliine. In order to examine the relationship between DNA adduct levels and the incidence of hemangiosarcomas, we have measured DHR-derived DNA adduct levels in purified rat and mouse liver endothelial cells, the cells of origin for the hemangiosarcomas. F344 rats and B6C3F1 mice were treated by gavage 5 days per week for 2 weeks with riddelliine at 1.0 mg/kg for rats and 3.0 mg/kg for mice. One, 3, 7, and 28 days after the last dose, liver parenchymal and endothelial cell fractions were isolated, and the quantities of DHR-derived DNA adducts were determined by 32P-postlabeling/HPLC. The DHR-derived DNA adduct levels in the endothelial cells were significantly greater than in the parenchymal cells. The DNA adduct levels in rat endothelial cells were greater than in the mouse endothelial cells. These results indicate that the levels of riddelliine-induced DNA adducts in specific populations of liver cells correlate with the preferential induction of liver hemangiosarcomas by riddelliine. PMID- 15127729 TI - A transparent drape aids abdominal aortic surgery. PMID- 15127730 TI - Combined insulation and smoke extraction for the diathermy blade. PMID- 15127731 TI - A novel technique for testicular tunnelling. PMID- 15127732 TI - Preparing digital images for publication. PMID- 15127733 TI - What to do if it gets "bigger". PMID- 15127734 TI - Erosive adenomatosis of the nipple--a report of three cases. PMID- 15127735 TI - Pancreatic debridement in a district general hospital--viable or vulnerable? PMID- 15127736 TI - Colds, international policy and inequalities. PMID- 15127737 TI - Multidisciplinary care for women with early breast cancer in the Australian context. PMID- 15127739 TI - Is SCHIP shipshape? Even successful insurance programs for kids are hard to keep going when budgets are squeezed. PMID- 15127738 TI - Vascular Ehlers-Danlos syndrome. AB - Vascular Ehlers-Danlos syndrome, also known as Ehlers-Danlos syndrome type IV, is a life-threatening inherited disorder of connective tissue, resulting from mutations in the COL3A1 gene coding for type III procollagen. Vascular EDS causes severe fragility of connective tissues with arterial and gastrointestinal rupture, and complications of surgical and radiological interventions. As for many rare orphan diseases, delay in diagnosis is common, even when the clinical features are typical, leading to inadequate or inappropriate treatment and management. In childhood many individuals with vascular EDS are first thought to have coagulation disorders. In adulthood, four main clinical findings, including a striking facial appearance, easy bruising, translucent skin with visible veins and rupture of vessels, gravid uterus or intestines, contribute to the diagnosis, which can be confirmed by SDS-PAGE studies of type III procollagen molecules synthesis by cultured fibroblasts or by the identification of a mutation in the COL3A1 gene coding for type III procollagen. Vascular EDS is inherited as an autosomal dominant trait. Varied molecular mechanisms have been observed and, of the mutations described to date, most have been unique to each family or "private", with no correlation between genotype and phenotype. Vascular EDS is of particular importance to surgeons, radiologists, obstetricians and geneticists since, although there is currently no specific treatment for the condition, knowledge of the diagnosis may help in the management of visceral complications, pregnancy and genetic counseling. PMID- 15127740 TI - David Weaver Robinson, M.D., 1914 to 2003. PMID- 15127741 TI - [Should varicoceles be treated, or not? When and how?]. PMID- 15127742 TI - [Should varicoceles be treated, or not? When and how?]. PMID- 15127743 TI - Reactive oxygen and nitrogen species: diagnostic, preventive and therapeutic values. PMID- 15127744 TI - Assessment of VIE image quality using helical CT angiography: in vitro phantom study. AB - The aim of this study is to quantify the effects of helical CT acquisitions parameters on the magnitude of three-dimensional stair-step artefacts, visualization of renal ostium and morphologies of suprarenal stents observed using virtual intravascular endoscopy. This was performed in a phantom of the human abdominal aorta with a stent graft in situ. Stair-step artefacts were quantified by measuring the standard deviation of signal intensity on surface shaded images and the influence of these artefacts on the visualization of arterial ostia and stent morphologies were assessed by three radiologists. The methodology may be used to optimise the CT system performance for helical CT angiography in aortic stent grafting. PMID- 15127745 TI - MR cholangiography 3D biliary tree automatic reconstruction system. AB - An algorithm for reconstructing magnetic resonance cholangiography (MRC) biliary structure is proposed. The processing of MRC data can be divided into four phases. In the first phase, the region of interest (ROI) containing the liver and biliary ducts is extracted from the original volume data based on human anatomy and B-spline curve. The second phase involves segmenting the biliary ducts from the region identified in the previous phase. Because the image of biliary portion is brighter than the liver, the segmentation is started by choosing the brightest pixel in the ROI as the seed for 3D region growing. This procedure could be executed many times, depending on the provided stopping condition. In the third phase, the segmented biliary duct regions are traced to construct the biliary tree. An automated 3D tracking algorithm is proposed for this phase. This 3D tracking algorithm estimates the coordinates of the points along the medial axis of each biliary duct branch. The cross sections associated with the points along the medial axis are also calculated approximately during the tracking process. The biliary tree data structure is constructed in this phase. The biliary tree is reconstructed and rendered in the last phase. Although the proposed algorithm takes a longer time compared with the conventional approach, the reconstructed biliary tree 3D structure can provide more clearly image. A concise representation for the biliary tree can be achieved with the proposed method and provide both quantitative and structural information for clinical reference. PMID- 15127746 TI - Three-dimensional reconstruction and fusion for multi-modality spinal images. AB - Medical diagnosis can benefit from the complementary information in different modality images. Multi-modal image registration and fusion is an essential task in numerous three-dimensional (3D) medical image-processing applications. Registered images are not only providing more correlative information to aid in diagnosis, but also assisting with the planning and monitoring of both surgery and radiotherapy. This research is directed at registering different images captured from Computed Tomography (CT) and Magnetic Resonance (MR) imaging devices, respectively, to acquire more thorough information for disease diagnosis. Because MR bone model segmentation is difficult, this research used a 3D model obtained from CT images. This model accomplishes image registration by optimizing the gradient information accumulated around the bony boundary areas with respect to the 3D model. This system involves pre-processing, 2D segmentation, 3D registration, fusion and sub-system rendering. This method provides desired image operation, robustness verification, and multi-modality spinal image registration accuracy. The proposed system is useful in observing the foramen and nerve root. Because the registration can be performed without external markers, a better choice for clinical usage is provided for lumbar spine diagnosis. PMID- 15127747 TI - Lymph node segmentation from CT images using fast marching method. AB - Accurate lymph node size analysis is important medically. This paper presents an improved fast marching method to perform semi-automatic segmentation for lymph node from CT images. In this work, we have incorporated the gray scale information of the target region into the fast marching speed term and have given a hard constraint for the stop criteria, instead of only using the spatial image gradient, to remedy the 'boundary leaking' problem of the traditional fast marching method. Various experimental results are provided to demonstrate the effectiveness of the proposed method. PMID- 15127748 TI - Image analysis techniques for characterizing disc space narrowing in cervical vertebrae interfaces. AB - Image analysis techniques are introduced for evaluating disc space narrowing of cervical vertebrae interfaces from X-ray images. Four scale-invariant, distance transform-based features are presented for characterizing the spacing between adjacent vertebrae. K-means and self-organizing map clustering techniques are applied to estimate the degree of disc space narrowing using a four grade (0-3) scoring system, where 0 and 3 represent normal spacing and significant narrowing, respectively. For a data set of 294 vertebrae interfaces, experimental results yield average correct grade assignment of greater than 82.10% for each of the four grades using a one grade window around the correct grade. PMID- 15127749 TI - Tracking deforming aortas in two-photon autofluorescence images and its application on quantitative evaluation of aorta-related drugs. AB - This paper describes a novel approach to an objective measurement of aorta samples of rats and a quantitative evaluation of aorta-related drugs. Two-photon fluorescence microscopy is used for recording image sequences of deforming aorta. Time sequence snake models are used to track the structural deformations of aorta walls caused by drug stimulation of the elastic lamina in the aorta. Several objective and quantitative biomarkers extracted from these models are used as diagnostic indicators. In a preliminary study, the technique was successfully used for evaluating the effect of a newly developed drug-human erythrocyte derived depressing factor quantitatively and objectively. PMID- 15127750 TI - Application of region-based segmentation and neural network edge detection to skin lesions. AB - This paper proposes two approaches to the skin lesion image segmentation problem. The first is a mainly region-based segmentation method where an optimal threshold is determined iteratively by an isodata algorithm. The second method proposed is based on neural network edge detection and a rational Gaussian curve that fits an approximate closed elastic curve between the recognized neural network edge patterns. A quantitative comparison of the techniques is enabled by the use of synthetic lesions to which Gaussian noise is added. The proposed techniques are also compared with an established automatic skin segmentation method. It is demonstrated that for lesions with a range of different border irregularity properties the iterative thresholding method provides the best performance over a range of signal to noise ratios. Iterative thresholding technique is also demonstrated to have similar performance when tested on real skin lesions. PMID- 15127751 TI - Reduction of respiratory motion artifacts in transverse relaxation rate (R2) images of the liver. AB - An empirical motion artifact suppression technique has been developed to reduce the respiratory motion artifacts in axial single spin-echo magnetic resonance (MR) images of the liver post-acquisition. The correction scheme is based on the observation that the dominant motion artifacts within abdominal MR images are ghosts that follow the profile and signal intensity of high signal intensity boundaries, such as those for the subcutaneous fat along the anterior abdominal wall. The technique is applied to the reduction of respiratory motion artifacts in a spin echo image series of the liver of an iron-loaded patient and of a manganese chloride phantom subject to respiratory motion. Subsequent improvements to transverse relaxation rate (R2) image analysis are then demonstrated on the motion-corrected spin echo images, illustrating the utility of the technique for application in the R2 image-based measurement and mapping of liver iron concentration. PMID- 15127752 TI - 3D airway segmentation via hyperpolarized 3He gas MRI by using scale-based fuzzy connectedness. AB - Computerized segmentation of 3D tracheobronchial tree is a necessary first step for subsequent registration and analysis of pulmonary airway and vascular magnetic resonance (MR) images obtained by using hyperpolarized 3Helium gas and Gadolinium. The scientific and clinical implications of acquiring these data on the tracheobronchial tree (for studying ventilation, V) and on the coinciding pulmonary arterioles (for studying perfusion, Q), is the next frontier for static and dynamic pulmonary MRI. In this paper, we report an airway segmentation method from 3He MR images based on the scale-based fuzzy connectedness approach. Incorporated in this method are the pre-processing steps of inhomogeneity correction and intensity standardization. The basic sequential steps in the proposed airway segmentation method are: (1) image acquisition, (2) radio frequency field inhomogeneity correction, (3) standardization of MR image intensity scale, (4) seed specification, (5) scale-based fuzzy connected segmentation of airways, and (6) thresholding and binarization. The majority of these steps are automatically executed; others allow interaction through a graphical interface provided in the 3DVIEWNIX software system, in which the algorithms are implemented. The method achieves an overall precision of about 98% in terms of the extent of overlap in repeated segmentations. Its level of accuracy can be described by a true positive volume fraction of about 98% (considering manual delineation as the surrogate of true delineation), and a false negative and positive volume fraction of about 1%. The total operator and computational time required per study are on the average 2 and 20 min. PMID- 15127753 TI - Robust automatic coregistration, segmentation, and classification of cell nuclei in multimodal cytopathological microscopic images. AB - The paper describes the key component of the Multimodal Cell Analysis approach, a novel cytologic evaluation method for early cancer detection. The approach is based on repeated staining of a cell smear. The correlation of features and data extracted from the different stains, and related to relocated individual cells, may yield a dramatic increase of diagnostic reliability. In order to utilise the technique, fully automatic, adaptive image preprocessing techniques need to be applied, which are described in this article: coregistration of multimodal images, segmentation, and classification of cell nuclei. The presented feasibility study shows both efficiency and robustness of all steps being high regarding medical image material, and it strongly supports clinical application. PMID- 15127754 TI - Anterior osteophyte discrimination in lumbar vertebrae using size-invariant features. AB - Radiologists often examine X-rays of cervical, thoracic and lumbar vertebrae for determining the presence of osteoarthritis and osteoporosis. For individual vertebra assessment, the boundary increasingly digresses from the general rectangular shape as the vertebra becomes less normal in appearance. For an abnormal vertebra, bony growths ('osteophytes') may appear at the vertebral comers, resulting in a change in the vertebra's shape. Image processing techniques are presented for computing size-invariant, convex hull-based features to highlight anterior osteophytes. Feature evaluation of 714 lumbar spine vertebrae using a multi-layer perceptron yielded normal and abnormal average correct discrimination of 90.5 and 86.6%, respectively. PMID- 15127755 TI - Caudal dysplasia sequence: severe phenotype presenting in offspring of patients with gestational and pregestational diabetes. AB - The association of maternal diabetes mellitus and congenital anomalies is well established. Children of insulin-dependent diabetic women have an increased risk of congenital malformations, especially major multiorgan defects. The cardiovascular, central nervous, gastrointestinal, genitourinary and musculoskeletal are the most affected body systems. Studies also show that offspring of women with gestational diabetes (specially those with fasting hyperglycaemia) tend to have higher rates of congenital anomalies. We report two cases of infants born to unrelated mothers: one with diabetes mellitus first detected during pregnancy (gestational diabetes) and the other with pregestational diabetes. Both infants had amelia of the lower limbs (suggestive of caudal dysplasia sequence), together with cardiovascular, skeletal, urinary and gastrointestinal defects. While pregestational diabetes seems to leave no doubt about its teratogenicity, the association of gestational diabetes and fetal/newborn malformations is still under discussion. Complete absence of the lower limbs has not been reported in association with gestational diabetes, but it may represent a spectrum of the caudal dysplasia sequence. The presentation of two cases with the same clinical phenotype of mothers with gestational and pregestational diabetes supports the evidence that gestational diabetes can be responsible for the development of the most severe form of the caudal dysplasia sequence. PMID- 15127756 TI - Vertically transmitted hypoplasia of the abdominal wall musculature. AB - The prune belly syndrome (OMIM 100100) is an association of bladder dilation with hypoplasia of the abdominal wall muscles. This malformation sequence is due to early urethral obstruction. We report a family with abdominal wall muscular hypoplasia as an isolated defect, not associated with the urethral obstruction sequence. The proband is a q3-year-old male who presented with abdominal wall laxity and severe constipation. His mother, maternal grandmother and younger brother had varying degrees of abdominal wall muscular deficiency and constipation. His mother's condition was aggravated by her 2 pregnancies. This family shows vertical transmission (compatible with autosomal dominant or mitochondrial inheritance) of the abdominal phenotype of prune belly sequence without any evidence of urinary tract or renal pathology. The expression in the sons may remain incomplete because abdominal distention due to pregnancy will not occur. PMID- 15127757 TI - Dysmorphic facial features in aspartylglucosaminuria patients and carriers. AB - The facial photos of 76 aspartylglucosaminuria (AGU) patients, 29 obligate carriers and 78 unrelated controls were evaluated for dysmorphic features to see whether this autosomal recessive lysosomal storage disease includes a dysmorphic facial gestalt in addition to the well-known age-related coarsening of the facies and whether the carrier status of AGU might have an effect on the facial features. A consistent dysmorphic gestalt with hypertelorism, a short and broad nose with round nares, simple often small ears with small or missing lobule and modest folding of helices, thick lips and a square shape of face, was found to be present long before the coarsening begins. Recognition of this pattern of facial features might help in the early diagnosis of AGU. Statistically, puffy eyelids were found to be significantly more frequent in AGU carriers than in controls. These findings support an earlier implication that AGU carrier status might have a slight influence on the phenotype. PMID- 15127758 TI - Skeletal manifestations in Ohdo syndrome: a case with bilateral patella dislocations. AB - We report a new case of Ohdo syndrome with bilateral patella dislocations where surgical intervention has been indicated. A review of the skeletal manifestations reported in the literature on Ohdo syndrome reveals that joint laxity and skeletal deformities are important aspects of the phenotype. PMID- 15127759 TI - Axial mesodermal dysplasia sequence: autopsy findings. AB - We describe an infant with the phenotype of the "axial mesodermal dysplasia spectrum" who had the oculo-auriculo-vertebral sequence and caudal dysgenesis. Postmortem studies identified anomalies of the middle ear ossicles, and muscles, in association with microtia and atresia of the external auditory canals, but no cerebral abnormalities. PMID- 15127761 TI - VACTERL-H with triphalangeal thumb and hypothyroidism in a female patient. AB - The association of vertebral, cardiac, renal, limb anomalies, anal atresia, tracheo-esophageal fistula (VACTERL) with hydrocephalus (VACTERL-H) has recently been described. Both X linked and autosomal recessive forms have been described which are mostly indistinguishable clinically. Here we report a female newborn infant with hydrocephalus detected antenatally, oesophageal atresia with tracheo oesophageal fistula, anal atresia, renal and vertebral anomalies, and triphalangeal thumb as well as primary hypothyroidism. PMID- 15127760 TI - A trisomy 2 fetus with severe neural tube defects and other abnormalities. AB - Examination of an abortus from a 13 week miscarriage revealed a fetus of around 9 weeks developmental age with multiple abnormalities including microcephaly, iniencephaly and encephalocele continuous with cervical and thoracic spina bifida, whose karyotype was subsequently shown to be 47,XY, + 2. PMID- 15127762 TI - Loose anagen hair syndrome associated with colobomas and dysmorphic features. AB - Loose anagen hair syndrome is an uncommon congenital disorder. It may occur in association with other syndromes and dysmorphic features. We report a girl who fulfilled the diagnostic criteria for this syndrome as proposed by Tosti (Arch Dermatol 2002, 138: 521-522). She also had several other anomalies including colobomas. We suggest that the combination of features in this patient may constitute a specific syndromic phenotype. PMID- 15127763 TI - How wide is the ocular spectrum of Delleman syndrome? AB - We describe a patient with cerebral and cutaneous features typical of oculocerebrocutaneous syndrome. The ocular anomalies observed have not been previously reported in patients affected with this syndrome. PMID- 15127764 TI - Frontonasal dysplasia, neuronal migration error and lymphoedema of limbs. AB - We describe a 15-month-old girl born to a normal family and with non consanguineous parents presenting with an unusual pattern of multiple congenital anomalies, including frontonasal dysplasia, neuronal migration error, lymphoedema of upper and lower limbs, and mild neuropsychomotor delay. Frontal and nasal haemangiomas, optic disc anomalies and hearing loss were also observed. This seems to be a previously undescribed syndrome of unknown aetiology. PMID- 15127765 TI - Klippel-Feil anomaly associated with thoracic hemivertebrae/butterfly vertebrae and patella hypoplasia. AB - A case with multiple congenital anomalies of the spine and bilateral patella hypoplasia is reported. To our knowledge, no report of concomitant Klippel-Feil syndrome, thoracic hemi vertebrae/butterfly vertebrae bilateral patella hypoplasia, reducible atlantoaxial instability, and occipitoatlantal fusion simultaneously has been reported in the literature. PMID- 15127766 TI - Femoral-facial syndrome with hemifacial microsomia and hypoglossia. AB - We present a boy diagnosed as femoral-facial syndrome with total agenesis of right lower limb, agenesis of femur and fibula of left lower limb and micrognathia, long philtrum as facial features. Some additional features were described as hemifacial microsomia, preauricular tags and hypoglossia. PMID- 15127767 TI - Pulmonary agenesis/hypoplasia, microphthalmia, and diaphragmatic defects: report of an additional case. AB - Diaphragmatic defects with associated pulmonary hypoplasia/agenesis and anophthalmia/microphthalmia have been reported as part of a complex syndromic phenotype. We report a case with the combination of these malformations. PMID- 15127768 TI - Unilateral pulmonary atresia with total sacral agenesis and other congenital defects. AB - Is there a common aetiologic element in the VACTERL syndrome and the caudal regression syndrome? We report a boy who fulfilled three criteria of the VACTERL association (he had thoracic hemivertebra, high anal atresia and tracheo oesophageal fistula) but also could be classified as caudal regression syndrome on the basis of the anal atresia and total sacral agenesis. We discuss the arguments in favour of both diagnosis. PMID- 15127769 TI - Acromegaloid facial appearance and hypertrichosis: a case suggesting autosomal recessive inheritance. AB - We report a patient with features of acromegaloid facial appearance syndrome and hypertrichosis. Her parental history suggests an autosomal recessive form of inheritance. PMID- 15127770 TI - Bilateral congenital longitudinal deficiency of the tibia associated with split hand and atrial septal defect. AB - We present the case of a 17-year-old boy with bilateral tibial hypoplasia and associated split hand and atrial septal defects. These have been reported previously to occur together by other investigators. There were no chromosomal abnormalities and the parents were non-consanguineous. PMID- 15127771 TI - Seckel syndrome associated with atrioventricular canal defect: a case report. AB - Seckel syndrome is a rare autosomal recessive disorder and its characteristic features are marked growth and mental retardation, significant microcephaly and a convex nose. We report a boy with this syndrome who also had severe cardiac anomalies. Although his parents were non-consanguineous, it is suggested that he had autosomal recessive inheritance. PMID- 15127772 TI - An infant with pentalogy of Cantrell and limb defects diagnosed prenatally. AB - We present a case of pentalogy of Cantrell which was diagnosed prenatally on routine ultrasound examination. There were several associated limb defects. We discuss the differential diagnosis and conclude that our case probably had a variant form of this syndrome. PMID- 15127774 TI - From surviving to thriving: the Beethoven factor. PMID- 15127773 TI - A newborn infant with left diaphragm agenesis, radial aplasia and preauricular appendices. AB - We report the case of a baby girl born to consanguineous parents who died 36 hours after birth. She had multiple preauricular appendices, right radial aplasia, triphalangeal thumb and several other anomalies. Differential diagnosis included Fryns syndrome, Pallister-Killian syndrome, and hemifacial microsomia. However, since our patient had congenital diaphragmatic hernia, a rare finding in hemifacial microsomia we suggest that she had a variant form of hemifacial microsomia or an undescribed new syndrome. PMID- 15127775 TI - Polarity addresses the "whole" in holistic. AB - Holistic healing modalities and integrative therapies are becoming a core part of primary care practices. This article will review the process of polarity therapy, identify the potential healing outcomes that can occur, and detail the elements it involves, including its energetic process, learn which clients might benefit from polarity therapy, and explore several of its health-building aspects. PMID- 15127776 TI - Healing ourselves, healing others: first in a series. AB - The healing role of nurses has been undervalued and undeveloped for a long time. A literature review of nurses and healing reveals the fears and frustration generations of women healers and nurses experienced as they strived to practice their art. The current nursing shortage reflects the toxicity of the nursing work environment. The satisfaction and liberation of nurses who have found ways to practice their calling also emerges, giving hope to those in the field as well as their patients. The first in a series, this literature review served as the foundation for a qualitative study entitled, "Medical-Surgical Nurses' Self Perceptions as Healers," which will appear in the next issue of Holistic Nursing Practice. PMID- 15127777 TI - "Quality time" serves families with a parent in recovery. AB - Families in which a parent or caregiver abuses chemicals face specific and unique problems that affect the health and well-being of the next generation, as well as the quality of current family life. This article reports on an intervention in which the multifamily group treatment method is used to address the needs of families with a parent in addiction recovery. Program participants reported continued sobriety at group's end and follow-up, increased knowledge and awareness of addiction as a disease, and increased closeness and better understanding among family members. PMID- 15127778 TI - Compassionate clinicians: take patient care beyond the ordinary. AB - This study sought to identify specific actions, interventions, and interpersonal relationships with patients exhibited by a group of compassionate healthcare clinicians. Researchers interviewed 24 hospital clinicians who were identified by administrators as being exemplary in caring and compassion. Analysis of qualitative data indicated that the clinicians do not attempt to distance themselves, but develop warm, empathic relationships with patients. Participating clinicians did not appear to sacrifice objectivity in practicing compassionate care, but were able to integrate the mind and heart in their work. Based on the qualitative analyses, researchers developed a preliminary model of affective clinician/patient interactions. PMID- 15127779 TI - Art and community health: lessons from an urban health center. AB - Staff at a nurse-managed urban health center conducted a series of art sessions to benefit the community. The authors believe the program's success clearly communicated the relationship between art and community health. As a result of the success of the sessions, plans are in the works to make art a permanent part of the health center's services. PMID- 15127780 TI - Mixing it up: compounding pharmacy creates individualized medications. PMID- 15127781 TI - Assessing methodological quality and biological plausibility in occupational health psychology. PMID- 15127782 TI - Is job strain a major source of cardiovascular disease risk? AB - Empirical studies on job strain and cardiovascular disease (CVD), their internal validity, and the likely direction of biases were examined. The 17 longitudinal studies had the highest validity ratings. In all but two, biases towards the null dominated. Eight, including several of the largest, showed significant positive results; three had positive, nonsignificant findings. Six of nine case-control studies had significant positive findings; recall bias leading to overestimation appears to be fairly minimal. Four of eight cross-sectional studies had significant positive results. Men showed strong, consistent evidence of an association between exposure to job strain and CVD. The data of the women were more sparse and less consistent, but, as for the men, most of the studies probably underestimated existing effects. Other elements of causal inference, particularly biological plausibility, corroborated that job strain is a major CVD risk factor. Additional intervention studies are needed to examine the impact of ameliorating job strain upon CVD-related outcomes. PMID- 15127783 TI - Urinary catecholamines and salivary cortisol on workdays and days off in relation to job strain among female health care providers. AB - OBJECTIVES: This study examined the effects of psychosocial job strain on the excretion of neuroendocrine stress hormones (adrenaline, noradrenaline, and cortisol) on workdays and days off. METHODS: Japanese female health care providers (N=16) filled out Karasek's job content questionnaire and had their neuroendocrine excretions (ie, urinary catecholamines and salivary cortisol) measured on a day off and on two workdays (one day shift and one night shift). After control for age and job experience as covariates, a repeated-measures analysis of variance was carried out. RESULTS: Noradrenaline excretion was significantly greater over time in the high-strain group than in the low-strain group, and that of the high-demand group was significantly greater over time than that of the low-demand group. Adrenaline excretion did not significantly differ between the groups. The group with high supervisory support had significantly higher adrenaline excretion than the group with low supervisory support. The concentration of salivary cortisol on a dayshift was significantly lower, but marginally, in the high-strain group than in the low-strain group. CONCLUSIONS: Psychosocial job strain is associated with greater noradrenaline excretion over time. This finding suggests unwinding sympathetic nervous activity. The low cortisol levels of the high-strain group may indicate circadian rhythm disturbance induced by job strain. Supervisory relationships may have a particular influence for the studied occupation because the participants had more administrative contact with supervisors than support at the worksite; therefore, supervisory support may increase adrenaline excretion. PMID- 15127784 TI - Occupational radiation dose estimation for Finnish aircraft cabin attendants. AB - OBJECTIVES: The objective of this study was to develop a method for assessing dose radiation on the basis of individual flight history and to estimate whether this method is applicable for cabin attendants without flight log data. METHODS: Questionnaire data were collected to determine attendants' flight history covering up to three decades. Finnair timetables and an expert panel of pilots were used to determine one to four representative flights in five route categories. The cumulative career and annual doses were calculated on the basis of the flight histories and route-specific exposure data. RESULTS: Questionnaire data were obtained from 544 flight attendants. The mean number of active workyears was 10.5 (range 0-30) years, and the mean cosmic radiation dose was 3.2 (range 0-9.5) mSv per active workyear. The mean cumulative career dose for all the cabin attendants was 34.0 (range 0-156.8) mSv. CONCLUSIONS: If no flight log data are available, survey data are needed for individual dose estimation when possible radiation effects on cabin crew are evaluated in epidemiologic studies. This method provides a crude procedure for assessing cosmic radiation exposure among attendants when survey data are missing. PMID- 15127785 TI - Building an evidence base for occupational health interventions. AB - This article summarizes arguments for building an evidence base for occupational health. Evidence is needed on the most effective ways of eliminating health hazards in the workplace and at work, enhancing healthy behavior or the empowerment of workers, and preventing and treating occupational diseases and occupational disability. An evidence base for occupational health must include systematic reviews. The Cochrane Collaboration has brought together some of the evidence; however, a search for systematic reviews on the top priorities in occupational health research showed that systematic reviews are lacking in many areas. Current reviewing methods can be adapted to the special features of occupational health. It is concluded that more effort should be invested in the preparation, maintenance, and dissemination of systematic reviews in order to create a necessary evidence base for occupational health interventions. Occupational health could benefit considerably from greater awareness of the evidence for and against various types of intervention. PMID- 15127787 TI - A novel hydrophilic support, CoFoam, for enzyme immobilization. AB - Lipases from different sources (porcine pancreas, Mucor miehei and Candida antarctica B) were covalently immobilized on a hydrophilic polyurethane composite (CoFoam). Their hydrolytic activities assayed with tributyrin were 0.55, 2.1 and 447 U g(-1), respectively. The activity of the C. antarctica B composite in the synthesis of methyl oleate in hexane was 8.8 U g(-1) compared to 60.6 U g(-1) for commercial Novozyme 435. The advantage of the CoFoam composite lies in the low pressure drop in a packed-bed reactor at fairly large flow rates. For example, at flow rates of 10-12 l min(-1), the pressure drop over 15 cm is typically 3 kPa. PMID- 15127786 TI - Applied biocatalysis for the synthesis of natural flavour compounds--current industrial processes and future prospects. AB - The industrial application of biocatalysis for the production of natural flavour compounds is illustrated by a discussion of the production of vanillin, gamma decalactone, carboxylic acids, C6 aldehydes and alcohols ('green notes'), esters, and 2-phenylethanol. Modern techniques of molecular biology and process engineering, such as heterologous expression of genes, site-directed mutagenesis, whole-cell biocatalysis in biphasic systems, and cofactor regeneration for in vitro oxygenation, may result in more biocatalytic processes for the production of flavour compounds in the future. PMID- 15127789 TI - Growth promotion of red pepper plug seedlings and the production of gibberellins by Bacillus cereus, Bacillus macroides and Bacillus pumilus. AB - The growth of red pepper plug seedlings was promoted by Bacillus cereus MJ-1, B. macroides CJ-29, and B. pumilus CJ-69 isolated from the rhizosphere. Gibberellins (GAs), a well-known plant growth-promoting hormone, were detected in the culture broth of their rhizobacteria. Among the GAs, the contents of GA1, GA3, GA4, and GA7, physiologically active GAs, were comparatively higher than those of others, suggesting that the growth promoting effect was originated from the GAs. This isthe first report on the production of GA5, GA8, GA34, GA44, and GA53 by bacteria. PMID- 15127788 TI - Biotransformation of fluorene, diphenyl ether, dibenzo-p-dioxin and carbazole by Janibacter sp. AB - Fluorene, diphenyl ether, dibenzo-p-dioxin, and carbazole were used by a dibenzofuran-utilizing Janibacter sp. strain YY-1. Metabolites were identified by GC-MS. Angular dioxygenation was the major pathway for degradation of fluorene, diphenyl ether, and dibenzo-p-dioxin but not for carbazole. Lateral dioxygenation of all tested compounds was indicated by the detection of mono- or di hydroxylated compounds. The bacterium also catalyzed the monooxygenation of fluorene at the C9 position. PMID- 15127790 TI - Design of novel analogues with potent antibiotic activity based on the antimicrobial peptide, HP(2-9)-ME(1-12). AB - To develop novel antibiotic peptides useful as therapeutic drugs, a number of analogues were designed to increase the hydrophobic helix region either by Trp substitution or net positive charge increase by Lys-substitution, from HP(2-9) ME(1-12). The antibiotic activities of these peptides were evaluated using bacterial (Salmonella tryphimurium, Proteus vulgaris, Bacillus subtilis and Staphylococcus aureus), fungi (Saccharomyces cerevisiae, Trichosporon beigelii and Candida albicans), tumor and human erythrocyte cells. The substitution of Lys for Thr at position 18 and 19 of HP(2-9)-ME(1-12) (HM5) increased activity against Proteus vulgaris and fungal strains without hemolysis. In contrast, substitution of Trp for Lys and Thr at positions 2, 15 and 19 of HP(2-9)-ME(1 12), respectively (HM3 and HM4), decreased activity but increased hemolysis against human erythrocytes. This suggests that an increase in positive charge increases antimicrobial activity whereas an increase in hydrophobicity by introducing Trp residues at C-terminus of HP(2-9)-ME(1-12) causes a hemolytic effect. Circular dichroism spectra suggested that the alpha-helical structure of these peptides plays an important role in their antibiotic effect but that the alpha-helical property is not connected with the enhanced antibiotic activity. PMID- 15127791 TI - Enzymatic synthesis of two novel non-reducing oligosaccharides using transfructosylation activity with beta-fructofuranosidase from arthrobacter globiformis. AB - Two novel non-reducing oligosaccharides together with tri- and tetra-saccharides were synthesized by transfructosylation activity from sucrose as a donor and cellobiose or cellotriose as an acceptor with a purified beta-fructofuranosidase from Arthrobacter globiformis IFO 3062, and these oligosaccharides were identified as O-beta-D-glucopyranosyl-(1-->4)-alpha-D-glucopyranosyl-(1-->2) alpha,beta-D-fructofuranoside and O-beta-D-glucopyranosyl-(1-->4)-beta-D glucopyranosyl-(1-->4)-alpha-D-glucopyranosyl-(1-->2)-alpha,beta-D fructofuranoside by spectrometric analyses. Both oligosaccharides were stable under condition at 100 degrees C for 30 min, and showed no degradation at pH 2. PMID- 15127792 TI - Stable hepatocyte transplantation using fibrin matrix. AB - Fibrin matrix, a naturally derived biodegradable polymer matrix, was evaluated as a scaffold for hepatocyte transplantation in an athymic mouse model. One week after transplantation, opaque conglomerates of the transplanted hepatocytes and fibrin matrix were found on the intestinal mesentery, whereas no transplanted hepatocytes were observed in control groups (transplantation of hepatocytes suspended in culture medium). The hepatocytes in the conglomerates retained hepatocyte-specific functions, as examined with histochemical and immunohistochemical stainings. Stable hepatocyte engraftment may thus be achieved by hepatocyte transplantation using fibrin matrix. PMID- 15127793 TI - Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper--Excel-based tool using pair-wise correlations. AB - The stability of standard gene expression is an elementary prerequisite for internal standardisation of target gene expression data and many so called housekeeping genes with assumed stable expression can exhibit either up- or down regulation under some experimental conditions. The developed, and herein presented, software called BestKeeper determines the best suited standards, out of ten candidates, and combines them into an index. The index can be compared with further ten target genes to decide, whether they are differentially expressed under an applied treatment. All data processing is based on crossing points. The BestKeeper software tool was validated on four housekeeping genes and 10 members of the somatotropic axis differentially expressed in bovine corpora lutea total RNA. The BestKeeper application and necessary information about data processing and handling can be downloaded on http://www.wzw.tum.de/gene quantification/bestkeeper.html. PMID- 15127794 TI - Biotransformation of lignin polymers derived from beech wood pulping by Sporobolomyces roseus isolated from leafy material. AB - The ability of the yeast, Sporobolomyces roseus, isolated from leafy material, to modify lignin derived from beechwood pulping was examined by FTIR and 13C NMR spectroscopy, which revealed oxidative cleavage of the Calpha-Cbeta linkages between lignin units. Using veratryl alcohol as a model substrate confirmed that Sp. roseus could oxidize veratryl alcohol into veratric acid. This yeast might be suitable for the pretreatment of lignocellulosic materials and/or for biotransformation of technical lignins. PMID- 15127795 TI - Scale-up micropropagation of sweet basil (Ocimum basilicum L.) in an airlift bioreactor and accumulation of rosmarinic acid. AB - Nodal explants with lateral buds and leaf-derived suspension cultures of sweet basil, Ocimum basilicum L., were cultured in 5 l airlift bioreactors for three weeks, thereby increasing the fresh wt of suspension cultures 2.5-fold. Rosmarinic acid accumulated at 29 micromicrog g(-1) dry wt in the suspension culture but, for micropropagated plants, it reached 178 microg g(-1) dry wt. PMID- 15127796 TI - Michael addition of imidazole with acrylates catalyzed by alkaline protease from Bacillus subtilis in organic media. AB - A new activity of alkaline protease from Bacillus subtilis for Michael addition reactions of imidazole, 4-nitro-1H-imidazole and 2-methyl-4-nitro-1H-imidazole with acrylates and acrylic acid was investigated. The reactions were carried out in pyridine at 50 degrees C for 72 h. Five N-substituted imidazole derivatives were obtained using acrylate esters, but not acrylic acid, in yields from 62% to 76%. PMID- 15127797 TI - Newly isolated bacterial strains belonging to Bacillaceae (Bacillus sp.) and Micrococcaceae accelerate death of the honey bee mite, Varroa destructor (V. jacobsoni), in laboratory assays. AB - Newly isolated bacterial strains belonging to Bacillaceae (Bacillus sp.), Micrococcaceae and three unidentified strains were tested for their pathogenicity against the mite, Varroa destructor. The Bacillus sp. strain and two of the strains belonging to the Micrococcaceae family significantly decreased the time for 50% mortality of the mite population (up to 57%) and hence may be potential control agents. In in vitro bioassay whole cells, extracellular broth and cellular extract of the Bacillus sp. strain effectively killed the mites, suggesting that both endotoxins and exotoxins contributed to the killing. PMID- 15127798 TI - Electric field-assisted biosorption. AB - A bisorption process using electric fields to facilitate contact between a sorbate and non-living biomass is described. The latter is enclosed within a semi permeable membrane together with an electrode. The counter electrode is placed in the sorbate solution and an established potential across the electrodes facilitates electrokinetic movement of the sorbate to the biosorbant material. PMID- 15127799 TI - Light sensitivity of Bifidobacterium longum in bioreactor cultivations. AB - Bifidobacteria are gaining commercial significance due to their probiotic properties. However, little is still known about the production of these bacteria and their behavior in bioreactors. Two Bifidobacterium longum strains were sensitive to light when grown in a transparent (glass) bioreactor under microaerophilic growth conditions (i.e. no gases added and slow mixing). The sensitivity was less clear the more anaerobic the initial conditions were. In a darkened bioreactor in microaerophilic conditions, the two strains grew with maximum specific growth rates of 0.36 h(-1) and 0.48 h(-1). In an illuminated bioreactor neither strain grew. In comparison, Lactobacillus reuteri was not sensitive to light under the same conditions. PMID- 15127800 TI - Flavor analysis in a pharmaceutical oral solution formulation using an electronic nose. AB - Flavors are commonly used in pharmaceutical oral solutions and oral suspensions to mask drug bitterness and to make the formulation more palatable. Flavor analysis during product development is typically performed by human organoleptic analysis, which is often expensive and less objective. A novel approach using a metal oxide sensor-based instrument (electronic-nose) for headspace analysis was explored to replace human sensory perception for consistent qualitative and quantitative analysis of flavors in a pharmaceutical formulation. The use of the electronic-nose technique to qualitatively distinguish among six common flavoring agents (raspberry, red berry, strawberry, pineapple, orange, and cherry) in placebo formulations was demonstrated. The instrument was also employed to identify unknown flavors in drug formulation placebos. Raspberry flavor samples from different lots made by the same manufacturer, as well as freshly prepared and aged samples, were also distinguished by electronic-nose. Therefore, the instrument can potentially be used for identity testing of different flavor raw materials and the flavored solution formulations. The electronic-nose was also employed successfully for quantitative analysis of flavors in an oral solution formulation. The quantitative method might be used to assay the flavor concentration during release testing of the oral solution formulation or to monitor flavor shelf-life in the marketed container. It can also be implemented for packaging selection for the formulation in order to ensure the flavor shelf life. Chemometric methodologies including principal component analysis (PCA), discriminant factorial analysis (DFA), and partial least squares (PLS), were used for data processing and identification. PMID- 15127801 TI - Development and validation of a high-resolution capillary electrophoresis method for multi-analysis of ragaglitazar and arginine in active pharmaceutical ingredients and low-dose tablets. AB - A selective, sensitive and robust capillary electrophoresis (CE) method has been developed and validated for multi analysis of ragaglitazar (also known as NNC 61 0029 or DRF 2725) and its counter ion arginine in Active Pharmaceutical Ingredients (API) and low-dose tablets (0.5, 1.0 and 2.0 mg). The method covers a total number of 12 analyses for the API and tablets: assay and identification of ragaglitazar and arginine, chiral purity of ragaglitazar and purity of ragaglitazar. After a simple extraction of samples with acetonitrile and 0.01 M sodium hydroxide (10:90), separation was performed using a combination of two cyclodextrins; sulfobutylether-beta-cyclodextrin (SB-beta-CD) and dimethyl-beta cyclodextrin (DM-beta-CD) in the electrolyte. The method showed good specificity and could separate and detect ragaglitazar, the distomer (the (+)-enantiomer) and arginine. The LOQ was found to be 0.10%, corresponding to 0.2 ng (0.5 microg/ml) for ragaglitazar and the distomer. Linearity was observed to be from 0.5 to 15 microg/ml (range 0.2-60 ng) and 400-600 microg/ml (range 1603-2404 ng) for ragaglitazar and 166-250 microg/ml (range 668-1000 ng) for arginine. The accuracy (as percent recovery) for ragaglitazar was found to be 101-106% (at 400-600 microg/ml), 101-125% (at 0.5-15 microg/ml) and for arginine 97-101% (at 166-250 microg/ml). The repeatability for the detection of peaks as R.S.D. was found to be as follows, ragaglitazar: 0.05%, distomer: 1.01%, largest single impurity: 5.84%, total impurities: 0.90% and arginine: 2.00%. The intermediate precision for the detection of peaks as R.S.D. was found to be as follows, ragaglitazar: 0.63%, distomer: 1.98%, largest single impurity: 5.22%, total impurities: 13.17% and arginine: 3.50. PMID- 15127802 TI - Simultaneous spectrophotometric determination of pseudoephedrine hydrochloride and ibuprofen in a pharmaceutical preparation using ratio spectra derivative spectrophotometry and multivariate calibration techniques. AB - Spectrophotometric methods are described for the simultaneous determination of pseudoephedrine hydrochloride and ibuprofen in their combination. The obtained data were evaluated by using five different methods. In the first method, ratio spectra derivative spectrophotometry, analytical signals were measured at the wavelengths corresponding to either maximums and minimums for both drugs in the first derivative spectra of the ratio spectra obtained by using each other spectra as divisor in their solution in 0.1 M HCl. In the other four spectrophotometric methods using chemometric techniques, classical least-squares, inverse least-squares, principal component regression and partial least-squares (PLS), the concentration data matrix were prepared by using the synthetic mixtures containing these drugs in methanol:0.1 M HCl (3:1). The absorbance data matrix corresponding to the concentration data matrix was obtained by the measurements of absorbances in the range 240-285 nm in the intervals with deltalambda = 2.5 nm at 18 wavelengths in their zero-order spectra, then, calibration or regression was obtained by using the absorbance data matrix and concentration data matrix for the prediction of the unknown concentrations of pseudoephedrine hydrochloride and ibuprofen in their mixture. The procedures did not require any separation step. The linear range was found to be 300-1300 microg/ml for ibuprofen and 100-1300 microg/ml for pseudoephedrine hydrochloride in all five methods. The accuracy and the precision of the methods have been determined and they have been validated by analyzing synthetic mixtures. The five methods were successfully applied to tablets and the results were compared with each other. PMID- 15127803 TI - Differentiation of bovine and porcine gelatins using principal component analysis. AB - Gelatin is a collagen derivative, which has a large application in the pharmaceutical, food and adhesive industries as well as photography. The large similarity in structure and properties of gelatins from different origins makes their differentiation difficult. Certain chemometric methods, such as principal component analysis (PCA), can help to classify and characterize gelatin components. In this study 14 bovine and 5 porcine gelatins were examined. The analysis procedure involved complete hydrolysis of samples by classic acid hydrolysis in order to release their amino acid residues. Separation and determination of amino acids was achieved by reversed-phase (RP) HPLC following pre-column derivatisation. Orthophtaldialdehyde (OPA) and 4-chloro-7-nitro benzofurazane (NBD-Cl) were used as derivatisation reagents. From the 20 peaks detected by HPLC analysis, one was very typical in bovine gelatin. Peak height, area, area percentage and width were used to make matrixes. Principal component analysis with the MATLAB program was used to differentiate these gelatins. PCA on matrix of height, width and total matrix were resulted in good differentiation between bovine and porcine gelatins. PMID- 15127804 TI - A HPLC validated assay of paclitaxel's related impurities in pharmaceutical forms containing Cremophor EL. AB - A HPLC method has been developed for the determination of the paclitaxel's related impurities in pharmaceutical forms. This method ensures the rapid determination of related impurities in the presence of polyoxyl castor oil--the main constituent of paclitaxel's clinical formulation vehicle. The method is simple and does not require any preliminary treatment of the sample. The method was fully validated. PMID- 15127805 TI - Rapid HPLC method for the determination of paclitaxel in pharmaceutical forms without separation. AB - A HPLC method has been developed for the in-process determination of the paclitaxel from dosage pharmaceutical forms. This method ensures the rapid determination of paclitaxel in the presence of polyoxyl castor oil--the main constituent of paclitaxel's clinical formulation vehicle. The method is simple and rapid and does not require any preliminary treatment of the sample. The method was fully validated. PMID- 15127806 TI - Adsorptive stripping voltammetric behaviour of azomethine group in pyrimidine containing drugs. AB - The stripping voltammetric behaviour of buspirone hydrochloride (BUS) and piribedil (PIR), as models of pyrimidine-containing compounds, was studied using a hanging mercury drop electrode (HMDE). A sensitive adsorptive stripping voltammetric method for determination of such drugs is described. The voltammetric peaks were obtained at -1.23 and -1.22 V for BUS and PIR. respectively, which correspond to the reduction of the azomethine group of pyrimidine ring in Britton-Robinson buffer (pH 7). Factors such as pH of supporting electrolyte, accumulation potential and time and instrumental parameters were optimized. Calibration plots and regression data validation, accuracy, precision, limits of detection, limits of quantification, and other aspects of analytical merit are presented. The applicability of the method was evaluated through determination of BUS and PIR in tablet dosage forms. A preliminary study of the analysis of plasma samples, spiked with the investigated drug, after a simple extraction procedure is described. PMID- 15127807 TI - Preparation and study the 1:2 inclusion complex of carvedilol with beta cyclodextrin. AB - An inclusion complex of beta-cyclodextrin with carvedilol was prepared by using a convenient new method of microwave irradiation. Phase-solubility studies demonstrated the ability of beta-cyclodextrins to complex with carvedilol and increase drug solubility. The structure of inclusion complex was determined by fluorescence spectroscopy and 1H NMR, 13C NMR measurements in solution. The solid inclusion was characterised by infrared spectroscopy, differential scanning calorimetry (DSC) and element analysis. These experimental results confirmed the existence of 1:2 inclusion complex of carvedilol with beta-cyclodextrin, the formation constant of complex was determined by the fluorescence method. Molecular modeling predicted the energy-minimized structure of the complex. PMID- 15127808 TI - High-performance thin-layer chromatography with a derivatization procedure, a suitable method for the identification and the quantitation of busulfan in various pharmaceutical products. AB - Busulfan is an alkylating agent widely used in combination chemotherapy regimens followed by allogeneic or autologous hematopoietic stem cell transplantation (HSCT). We present a rapid method for assaying busulfan in pharmaceutical preparations using high-performance thin-layer chromatography (HPTLC) and derivatization with 4-nitrobenzylpyridine. The method is accurate and precise and allows quantitation of busulfan in aqueous solutions from 100 to 500 microg/ml. It is suitable for identification. quantitation and stability studies of busulfan in pharmaceutical products, i.e. capsules or infusion bags prepared in a hospital pharmacy. PMID- 15127809 TI - Data preprocessing by wavelets and genetic algorithms for enhanced multivariate analysis of LC peptide mapping. AB - Peptide mapping by means of liquid chromatography is a powerful technique used for the characterisation and analysis of the primary structure of proteins. Subtle changes in the covalent structure of the protein can be detected by means of the chromatographic profile (fingerprint). Chromatographic methods, however, display variations in the chromatographic profile even at identical instrumental settings and sample conditions. These variations may be due to changes of the chromatographic conditions, e.g. slight shifts in column temperature, and degradation or alterations of the stationary phase or small changes in the trifluoroacetic acid (TFA) concentration. Such variations may result in varying retention times and peak shapes of the analytes and differences in the chromatographic baseline, thereby having a detrimental impact on the results obtained on multivariate analysis of peptide maps. In order to reduce the non sample-related variations and to be able to more fully extract the information in peptide mapping, approaches for achieving this objective are outlined in the present study. These methods are denoising and data compression of the chromatograms by wavelets, baseline corrections by linear interpolation, and peak shift alignments towards a target chromatogram by means of a genetic algorithm. Visual inspections of preprocessed chromatograms and principal component analysis (PCA) score plots demonstrate the efficiency of the methodology used. Furthermore, deliberately added changes, e.g. insertions of small Gaussian peaks (outliers), are more easily detected by the proposed methods than from the original chromatograms by multivariate analysis. PMID- 15127810 TI - Determination of vitamin B6 in pharmaceutical formulations by flow injection solid phase spectrophotometry. AB - In this work, a new solid phase spectrophotometric method in association with flow injection analysis for Vitamin B6 (pyridoxine) determination has been developed with direct measurement of light-absorption in C18 material. In the developed method, successive passage of the complex, previously formed in the flowing stream, and eluent through the flow cell and continuous monitoring of the process provided the analytical information needed to determine pyridoxine. Pharmaceutical samples containing Vitamin B6 were previously dissolved in 0.1 mol l(-1) phosphate buffer solution (pH 7.5) and a sample volume of 235 microl was injected directly into carrier stream consisting of a mixture of methanol and 0.1 mol l(-1) phosphate buffer solution adjusted to pH 7.0 (1+1, v/v). The blue indophenol dye produced from the reaction between pyridoxine and N,N-diethyl-p phenylenediamine after oxidation by potassium hexacyanoferrate(III) was quantitatively retained on C18 support and the spectrophotometric detection was performed simultaneously at 633 nm. The retained complex was quickly eluted from C18 material with the eluent stream consisting of a mixture of methanol and 0.01 mol l(-1) HCl (6+4, v/v). The results showed that the proposed method is simple, rapid and the analytical response is linear in the concentration range of 0.5-10 and 0.2-4 mg l(-1) using 235 and 860 microl of sample, respectively. The limits of detection are 0.15 and 0.060 mg l(-1) and the R.S.D. are 3.6% (at 2 mg l(-1) level) and 4.0% (at 1 mg l(-1) level) using sample volume of 235 and 860 microl, respectively. The system presented an analytical throughput of 15 determinations per hour when a sample volume of 235 microl was utilized. The procedure was successfully applied to the determination of Vitamin B6 in pharmaceutical formulations containing vitamins of B group and others active principles such as Vitamin C and minerals. PMID- 15127811 TI - Successive determination of thiamine and ascorbic acid in pharmaceuticals by flow injection analysis. AB - A simple and rapid fluorimetric method for the determination of mixtures of thiamine and ascorbic acid is proposed. The procedure is based on the oxidation with mercury(II) of the B1 and C vitamins to form thiochrome (TC) and quinoxaline derivate, respectively. Both reaction products exhibit fluorescence at the same wavelengths (lambdaex = 356 and lambdaem = 440 nm). The procedure is optimised in a flow injection (FI) system and applied with excellent results in the determination of B1 and C vitamins in commercial pharmaceutical preparations. The calibration graphs were linear over the range 2-100 microg ml(-1) for thiamine and 5-100 microg ml(-1) for ascorbic acid. The throughput was 25 samples per hour. PMID- 15127812 TI - Isolation, structural elucidation and in vitro activity of 2-acetyl-2 decarboxamido-oxytetracycline against environmental relevant bacteria, including tetracycline-resistant bacteria. AB - 2-Acetyl-2-decarboxamido-oxytetracycline (ADOTC) is a major impurity of oxytetracycline (OTC) produced as a side product during fermentation. ADOTC was isolated from OTC and other impurities using preparative HPLC. The preparative column was an Xterra MS, C18 chromatographic column (100 mm x 19 mm i.d., 5 microm), and the mobile phase contained methanol-water (27:73 (v/v)) with 0.08 M formic acid added. The flow rate was 9.0 ml/min. It was possible to isolate few milligram ADOTC in a day. The compound was unambiguously identified using NMR and MS-MS. The anti-microbial activity against activated sludge bacteria was determined giving a potency of only 3% of that of OTC. With tetracycline resistant bacteria, no anti-microbial activity was observed, indicating a mode of action similar to that of OTC. PMID- 15127813 TI - Metal content monitoring in Hypericum perforatum pharmaceutical derivatives by atomic absorption and emission spectrometry. AB - Metals have been investigated in different plant materials in order to establish their normal concentration range and consider their role in plants as part of human medicinal treatment. Metal monitoring as a pattern recognition method is a promising tool in the characterization and/or standardization of phytomedicines. In the present work measurable amounts of Ca, Cu, K, Li, Mg, Mn, Na, Ni, and Zn were detected in phytopharmaceutical derivatives of Hypericum perforatum by atomic techniques. Atomic methodologies like flame atomic absorption spectrometry (FAAS) and electrothermal atomic absorption spectrometry (ETAAS) allow reliable determination of mineral content in pharmaceutical quality control of medicinal plants. Additionally, capillary electrophoresis (CE) patterns of characteristic components (fingerprints) have been performed for the search of adulterants in phytopharmaceutical products. PMID- 15127814 TI - Simultaneous extraction and quantitation of fentanyl and norfentanyl from primate plasma with LC/MS detection. AB - The quantitation of both fentanyl and its desalkyl metabolite, norfentanyl, in plasma using LC/MS has not been previously described. The detection and quantitation of fentanyl and norfentanyl was achieved using LC/MS detection. The liquid-liquid extraction used toluene as the organic phase. Chromatography was carried out using a Zirchrom-PBD (50 mm x 2.1 mm, 3 microm) column with a mobile phase of acetonitrile-ammonium acetate (10 mM), citrate (0.1 mM, pH 4.4) (45:55, v/v) with a flow rate of 0.3 ml/min. Mass spectroscopy detection was performed using ESI in the positive mode. The LOQ for fentanyl was 25 pg/ml and norfentanyl was 50 pg/ml. For the concentrations of 75, 250, and 750 pg/ml, respectively, fentanyl had inter-day precisions of 6.6, 7.2, and 6.6% with accuracies of 4.0, 5.1, and 5.1% and intra-day precisions of 1.6, 1.9, and 1.9% with accuracies of 11.6, 9.4, and 8.4%, and norfentanyl had inter-day precisions of 7.4, 0.3, and 0.7% with accuracies of 9.1, 8.8, and 12.3% and intra-day precisions of 5.3, 1.4, and 0.1% with accuracies of 10.9, 8.9, and 12.8%. The recoveries of fentanyl were 85, 92, and 75% and of norfentanyl were 40, 49, and 46% at the 75, 250, and 750 pg/ml concentrations, respectively. PMID- 15127815 TI - The quantification of paracetamol, paracetamol glucuronide and paracetamol sulphate in plasma and urine using a single high-performance liquid chromatography assay. AB - A range of analytical methods exist for the determination of paracetamol in biological fluids. However, to understand the fate of paracetamol and the effect of other drugs on its disposition in vivo, the major metabolites require quantification in urine and plasma. A method to simultaneously quantify paracetamol, paracetamol glucuronide (PG) and paracetamol sulphate (PS) in plasma and urine with superior sensitivity is therefore desired, especially if the volume of plasma available is low. A simple isocratic reverse phase high performance liquid chromatography (HPLC) assay with spectrophotometric detection has been developed. The method, requiring only 100 microl of plasma and 50 microl of urine, utilizes a reversed-phase C18 column, a wavelength of 254 nm for detection and a mobile phase composed of potassium dihydrogen orthophosphate (0.1 M)-isopropanol-tetrahydrofuran (THF) (100:1.5:0.1, v/v/v) adjusted to pH 3.7 with phosphoric acid. The method is sensitive and linear in plasma within a concentration range from 0.4 to 200 microM for paracetamol, PG and PS. For PG and PS in urine, the method is sensitive and linear within a concentration range from 100 to 20,000 microM. Over these ranges, accuracy and precision were less than 12%. The assay has been used to measure concentrations of paracetamol and the two metabolites in plasma collected by finger-prick sampling and of the metabolites in urine from healthy volunteers administered a single oral dose of 1000 mg of paracetamol. PMID- 15127816 TI - Analysis of benzodiazepines in dynamically coated capillaries by CE-DAD, CE-MS and CE-MS2. AB - The applicability of a low pH volatile electrolyte for fast analysis of benzodiazepines with CE-MS was investigated. The electrolyte is based on a commercially available CEofix buffer system that produces a substantial and highly reproducible electroosmotic flow through a dynamic double coating principle. The system was first evaluated with a mixture of benzodiazepine standards in CE-DAD and the electrolyte composition was further optimized for CE MS. The LOD for the six selected benzodiazepines with CE-MS was ca. 100 ppb, except for diazepam, for which the LOD was lower than 50 ppb. RSDs varied from 0.51 to 1.02% (n = 7) for migration times and from 4.75 to 11.80% (n = 7) for peak areas. The method was successfully applied to the analysis of a spiked urine sample after solid-phase extraction (SPE). CE-MS2 was performed on a standard mixture. PMID- 15127817 TI - The use of 96-well Scintiplates to facilitate definitive metabolism studies for drug candidates. AB - Semi-quantitative analysis of the drug-related components in biological samples collected during definitive metabolism studies using radiolabelled drug candidates is commonly achieved by HPLC profiling, using either on-line radiochemical detection or off-line liquid scintillation counting (LSC) following collection of the HPLC eluent into vials. However, although the use of LSC with vials has high sensitivity, the approach is time-consuming, laborious and destructive, whilst on-line detection methods are inappropriate for samples with low-levels of radioactivity (commonly the case with plasma samples). The use of 96-well microtitre plates (Scintiplates) for fraction collection during HPLC profiling provides a sensitive, effective and efficient alternative method for the semi-quantitative analysis of radiolabelled components in biological samples. Furthermore, the approach is non-destructive, such that subsequent identification of the isolated components can be achieved. Although the Scintiplate methodology is not appropriate for the analysis of excreta samples, where quenching of the radiochemical signal by endogenous components was observed, the approach was demonstrated to be valid for the relative quantification of [14C]-labelled material in plasma samples for all species investigated. In addition, good sensitivity was observed, with a counting efficiency of 79% for [14C], such that a drug-related component accounting for 10-15 dpm is quantifiable. The utility of the methodology for profiling circulating metabolites was demonstrated by the analysis of a rat plasma sample following oral administration of [14C]-UK 349,862. The Scintiplate approach and subsequent mass spectrometric analysis resulted in the relative quantitation and specific characterisation of circulating metabolites accounting for 93% of the total plasma radioactivity. PMID- 15127818 TI - Determination of aplidin, a marine-derived anticancer drug, in human plasma, whole blood and urine by liquid chromatography with electrospray ionisation tandem mass spectrometric detection. AB - A sensitive and highly specific liquid chromatographic method with electrospray ionisation tandem mass spectrometric detection (LC-ESI-MS/MS) is reported for the determination in human plasma, whole blood and urine of Aplidin (APL), a novel depsipeptide derived from the tunicate Aplidium albicans with a potent cytotoxic activity under investigation in clinical studies. Didemnin B was used as internal standard and, after protein precipitation with acetonitrile and liquid-liquid extraction with chloroform, APL was separated by liquid chromatography using a reversed-phase C18 column and a linear gradient of acetonitrile in water (both containing 0.5% formic acid). Detection was performed using a turboionspray source operated in positive ion mode and by multiple reaction monitoring (MRM; m/z 1111 --> 295 for APL and m/z 1113 --> 297 for didemnin B). The method was linear (r > or = 0.9933) over the range 1-250 ng/ml, with intra- and inter-batch precision and accuracy below 12.2% (except at LLOQ < or = 15.4%) for both plasma and urine. Recoveries were moderate, ranging from 54 to 70% in plasma and blood, and from 46 to 60% in urine, for both APL and didemnin B. The LOD was 0.25 ng/ml for both matrices. APL resulted stable in the different matrices at least for 6 h (both at room temperature and 37 degrees C), after freeze and thaw cycles and long term storage at -20 degrees C. The method allowed demonstrating that APL is in a dynamic equilibrium between plasma and blood cells. Moreover, the method was successfully applied to the pharmacokinetic study of Aplidin in cancer patients. PMID- 15127819 TI - Quantitative analysis of squalamine, a self-ionization-suppressing aminosterol sulfate, in human plasma by LC-MS/MS. AB - The special physico-chemical property of squalamine enables the formation of intra- or inter-molecular non-volatile strong salt, which is difficult to ionize in a mass spectrometer's interface. A sensitive, accurate, precise, and specific method for the quantitative determination of this self ion-suppressing compound in human plasma has been developed and validated using high performance liquid chromatography (HPLC) coupled with positive electrospray tandem mass spectrometry (MS/MS). Solid phase extraction (SPE) technique was utilized to extract human plasma samples using the Waters Oasis HLB cartridges. Deuterated squalamine was used as the internal standard (IS). Positive multiple reaction monitoring (MRM) mode was used to achieve both sensitivity and selectivity. A quadratic linearity range over 5-1000 ng/ml, R > 0.999 was achieved. Performance of the method has been validated and met all the specifications set forth in the US Food and Drug Administration's May 2001 "Bioanalytical Method Validation Guidance for Industry". Different sample reconstitution solutions were found to have dramatic impact on sensitivity of mass spectrometer used to squalamine. This is the first quantitation method using a positive and true multiple reaction monitoring mode detection for squalamine. PMID- 15127820 TI - High performance liquid chromatography with fluorescence detection for the determination of phenylpropanolamine in human plasma and rat's blood and brain microdialysates using DIB-Cl as a label. AB - A high performance liquid chromatographic method for the determination of phenylpropanolamine (PPA) in human plasma and rat's brain and blood microdialysates using fluorescence (FL) detection after precolumn derivatization with 4-(4,5-diphenyl-1H-imidazole-2-yl)benzoyl chloride (DIB-Cl) is described. PPA was extracted from plasma samples by a liquid-liquid extraction method with ethyl acetate followed by derivatization with DIB-Cl, while the blood and brain microdialysates were directly subjected for derivatization. The DIB-derivatives of PPA and the internal standard, ephedrine (EP), were then separated using an isocratic HPLC-FL set at excitation and emission wavelengths of 325 and 430 nm, respectively, on an ODS column. Calibration curves of PPA in spiked human plasma were linear over the concentration range of 5-5000 nM (0.755-755 ng/ml) and those in spiked blood and brain microdialysates were linear over the range of 25-5000 nM (3.775-755 ng/ml) with limits of detection of 17, 48 and 40 fmol on column in plasma and blood and brain microdialysates, respectively. As well, the intra- and the inter-assay precisions were lower than 12% for human plasma and the microdialysates. The method was successfully applied for the monitoring of PPA levels in rat's brain and blood microdialysates administered with a single oral dose of PPA (2.5 mg/kg). PMID- 15127821 TI - Quantitative analysis of lincomycin in animal tissues and bovine milk by liquid chromatography electrospray ionization tandem mass spectrometry. AB - A sensitive method for determining lincomycin in bovine milk, animal muscles and organs using liquid chromatography electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS) is presented. Milk and homogenized animal tissues were extracted with acetonitrile twice after addition of an appropriate amount of clindamycin, a lincosamide analogue as the internal standard. The combined extracts were finally made up to 10 ml with distilled water and partitioned with hexane to remove the animal fats prior to analysis. Analytes in the extracts were separated on a reversed phase C18 column (250 mm x 2.1 mm, 5 microm) using a mobile phase of a 3:7 (v/v) mixture of 0.1% formic acid in acetonitrile and an ammonium formate buffer (ammonium formate:formic acid:acetonitrile:water, 1:5:50:950, v/v/v/v) running at a flow rate of 0.2 ml min(-1). Presence of lincomycin was confirmed by the presence of two characteristic product ions at m/z 126.1 and 359.2 within a defined retention time window from the precursor ion at m/z 407.2, whilst quantification was based on the relative ratio of the sum of the peak areas at m/z 126.1 and 359.2 for lincomycin to that of the internal standard (peaks at m/z 126.1 and 377.2) with reference to the respective ratios of the calibration standards. The validated method that was found to have linear responses in the calibration range from 25 to 3000 microg kg(-1) and satisfactory intra-day and inter-day accuracy (94.4-107.8%) and precision (1.3-7.8%) at concentrations ranging from 100 to 1500 microg kg(-1) has been applied to real samples and matrix spiked samples. It is considered robust and suitable for analysis of lincomycin in milk and animal tissues. PMID- 15127822 TI - Sensitive determination of erdosteine in human plasma by use of automated 96-well solid-phase extraction and LC-MS/MS. AB - A sensitive and selective method for quantitation of erdosteine in human plasma was established by use of 96-well solid-phase extraction (SPE) and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI/MS/MS). Plasma samples were transferred into 96-well OASIS HLB extraction plate using an automated sample handling system and the drugs were eluted with methanol. The eluents were then evaporated and reconstituted with mobile phase. All sample transfer and SPE was automated through the application of both the Perkin-Elmer MultiPROBE II HT and TOMTEC Quadra 96 workstation. Compounds were separated on a C18 column with 1 mM ammonium acetate-acetonitrile (80:20, pH 3.2), as mobile phase at a flow rate of 0.3 ml/min. The limit of quantitation (LOQ) was 0.2 ng/ml, using a sample volume of 0.2 ml for the analysis. The reproducibility of the method was evaluated by analyzing three at 14 quality control (QC) levels over the nominal concentration range from 0.2 to 5000 ng/ml. The intraday accuracy was found to range from 99.6 to 105.0% with precision (% RSD) of less than 4.76% at five QC levels. The interday accuracy was found to range from 95.0 to 100.5% with precision of less than 5.26% at five QC levels. Erdosteine produced a protonated precursor ion ([M+H]+) at m/z 250, and a corresponding product ion at m/z 204. Internal standard (letosteine) produced a protonated precursor ion ([M+H]+) at m/z 280 and a corresponding product ion at m/z 160. The high sample throughput of the method has been successfully applied to a pharmacokinetic study of erdosteine in human plasma. PMID- 15127823 TI - Development and validation of fluorescent receptor assays based on the human recombinant estrogen receptor subtypes alpha and beta. AB - This article describes the development and validation of two fluorescent receptor assays for the hRec-estrogen receptor subtypes alpha and beta. As a labelled ligand an autofluorescent phyto-estrogen (coumestrol) has been used. The estrogen receptor (ER) belongs to the nuclear receptor family, a class of soluble DNA binding proteins, mainly present in the cytoplasm of the cell, that act as ligand activated enhancer factors. It consists of two different forms, expressed as ER alpha (66 kDa) and ER-beta (59 kDa). The ER-alpha is mainly located in the uterus and the ER-beta can be found in vascular tissue. Detection and identification of compounds having estrogenic effects is of importance in drug discovery programmes within the pharmaceutical industry for their search for ER-subtype selective (ant)agonists which may prove to be of therapeutic value in treating a variety of estrogen-linked pathologies (breast cancer, osteoporosis, cardiovascular disease, type II diabetes and Alzheimer disease). Furthermore, interactions of (xeno )estrogens with the endogenous hormonal system of the exposed organism can affect embryos, gonads, and reproductive behaviour. The latter can eventually lead to reduced reproduction and deterioration of a population. For that reason, monitoring of (xeno-)estrogens in food products and in the environment, attracts considerable attention by health councils throughout the world. The following characteristics were obtained for the human recombinant (hRec) estrogen receptor beta assay, which is suitable for ER subtype selective drug-discovery purposes (IC50 values for 17-beta-estradiol and genistein were 5.1 nM and 25 nM, respectively): goodness of fit (R2) was always > 0.98 (x = 0.9933, n = 10). LLOQ of the assay is typically > or = 500 picomolar, whereas the ULOQ of the assay is < or = 20.0 nanomolar. For the hRec-estrogen receptor-alpha assay, which is suitable for monitoring of (xeno-)estrogens (IC50 values for 17-beta-estradiol and genistein were 0.68 nM and 65 nM, respectively) the following characteristics were obtained: goodness of fit (R2) was always > 0.96 (x = 0.9838, n = 10). LLOQ of the assay is typically > or = 200 picomolar, whereas the ULOQ of the assay is < or = 5.0 nanomolar. PMID- 15127824 TI - Rapid determination of telmisartan in pharmaceutical preparations and serum by linear sweep polarography. AB - The polarographic behaviors of telmisartan (TE) are investigated in 0.8 mol/l NH3.H2O-NH4Cl (pH = 8.9) supporting electrolyte. The results demonstrated that the reduction peak is obtained at ca. -1.30V, which corresponds to a catalytic hydrogen wave. Based on the catalytic hydrogen wave, a novel method has been developed for the determination of telmisartan by linear sweep polarography. Calibration curve is linear in the range 2.0 x 10(-7) to 3.0 x 10(-6) mol/l and the detection limit is 1.0 x 10(-7) mol/l. The proposed method is applied to the rapid determination of the telmisartan in capsule forms and biological sample without pre-separation. PMID- 15127825 TI - Chiral separation of tamsulosin isomers by HPLC using cellulose tris (3,5 dimethhylphenylcarbamate) as a chiral stationary phase. AB - A high-performance liquid chromatographic (HPLC) method was developed for the chiral separation of an antagonist of alpha1A adrenoceptors, tamsulosin and its S isomer. Baseline separation of the isomers was achieved within 35 min on a CHIRALCEL OD-RH column with a binary solvent mixture of 50 mmol l(-1) KPF6 acetonitrile (v/v (70:30), pH 5.0) as the optimized mobile phase. The detection limits and quantification limits of both R-isomer and S-isomer were 0.11 and 0.44 ng, respectively. The R.S.D. values of peak-area for the two isomer were 0.42% (of peak-height: 0.77%) for R-isomer and 0.64% (of peak-height:0.92%) for S isomer (n = 5). PMID- 15127826 TI - Simultaneous analysis of theanine, chlorogenic acid, purine alkaloids and catechins in tea samples with the help of multi-dimension information of on-line high performance liquid chromatography/electrospray-mass spectrometry. AB - A reverse phase high performance liquid chromatography (RP-HPLC) separation coupled with photo diode array detection (DAD) and electrospray ionization mass spectrometry (ESI-MS) detection was established for the analyzing of multiple bioactive compounds in tea and tea extracts. Theanine, chlorogenic acid, purine alkaloids and catechins were identified with authentic standard compounds and with MS-spectra. The content of theanine and catechins was measured by employing DAD and caffeine, chlorogenic acid, theobromine and theopylline by protonated molecular ion on selective ion recording (SIR) mode. The unity of LC/ESI-MS provides more qualitative and quantitative information comparing with general HPLC in the analysis of multi-components in tea, and complex extraction or sample pretreatment is unnecessary. The chromatogram acquired by using this method can be used as a bioactive components fingerprint for the quality control of tea and its extracts. With the help of multi-dimension information of HPLC-DAD-ESIMS, the compounds owning different chemical structure such as amino acid, catechins, etc. in tea and its extracts could be identified and determined in one run successfully. PMID- 15127827 TI - Identification and determination of the major constituents in traditional Chinese medicine Si-Wu-Tang by HPLC coupled with DAD and ESI-MS. AB - An HPLC/DAD/ESI/MS method was established for the qualitative and quantitative analysis of the major constituents in Si-Wu-Tang, a traditional Chinese medicine formula. Based on the baseline chromatographic separation of most constituents in Si-Wu-Tang on hypersil C18 column with water-acetonitrile-acetic acid as mobile phase, 12 compounds including phenolic acids, phthalides and terpene glycoside were identified by online ESI-MS and the comparison with literature data and standard samples. Most of these compounds derive from Paeonia lactiflora and Ligusticum chuanxiong. Seven of them were quantitated by HPLC coupled with DAD. The validation of the method, including sensitivity, linearity, repeatability, recovery, were examined. The linear calibration curve were acquired with R2 > 0.99 and LOD (S/N = 3) were between 0.75 and 5 ng. The repeatability was evaluated by intra- and inter-day assays and R.S.D. value were within +/- 2.38%. The recovery rates of selected compounds were in the range of 96.64-105.21% with R.S.D. less than 3.22%. PMID- 15127828 TI - Pneumococcal polysaccharide vaccines do not protect the elderly from pneumococcal infections. PMID- 15127829 TI - Pneumococcal vaccination for the elderly in The Netherlands? Assessment of the quality and content of available comparative studies. AB - BACKGROUND: A question that is currently topical in the Netherlands is whether it makes sense to introduce on a national scale vaccination against pneumococcal infections for elderly people who are at present receiving the influenza vaccination. We recently studied the scientific literature on the subject in an attempt to answer this question. METHODS: We searched for systematic reviews (SRs), randomised clinical trials (RCTs) and cohort studies in MEDLINE, EMBASE, the Cochrane Library, Current Controlled Trials and via Google (period 1966 to June 2002). The SRs and RCTs were assessed with a methodological checklist. RESULTS: We identified four SRs, two trials (of which one was pseudo-random) and one retrospective cohort study. The methodological quality of the SRs was reasonable and in this respect differed little among themselves. The SRs differed strongly with regard to subgroups, outcome measures, valency of vaccines, duration of follow-up and combination with influenza vaccination. The SRs showed that vaccination has more effect in low-risk groups, does not appear to be effective in high-risk patients and the elderly and is more effective in nonindustrialised countries. The outcomes based on the various outcome measures showed major differences. The three studies into the effectiveness of the pneumococcal vaccination in the elderly all showed major methodological shortcomings. For the majority of outcome measures the outcomes were negative. CONCLUSION: There is insufficient convincing evidence in favour of the introduction of the pneumococcal vaccination as a supplement to influenza vaccination for the elderly. It seems as if (international) opinion had already been fully formed before published studies and systematic reviews become available in the last few years. It is perhaps worth considering setting up a prospective trial in the elderly Dutch population. PMID- 15127830 TI - Prevention of infections in hyposplenic and asplenic patients: an update. AB - Patients with functional or anatomic asplenia are at a significantly increased risk of overwhelming infection, particularly involving the encapsulated bacteria Streptococcus pneumoniae and Haemophilus influenzae. The risk is highest in infants and young children, but adults also have an increased risk of infection. Preventive strategies are very important and fall into three major categories: immunoprophylaxis, antibiotic prophylaxis and education. Studies have shown that many asplenic patients are unaware of their increased risk for serious infection and the appropriate health precautions that should be undertaken. In this article we emphasise the need for preventive measures in hyposplenic and asplenic patients. We discuss the value of newly developed conjugate vaccines and the need for revaccination. Finally we draw up a recommendation for the preventive management in functional and anatomical asplenic patients. PMID- 15127831 TI - The StethoDop: a Doppler stethoscope attachment for investigation of arterial and venous insufficiency of the lower extremities. AB - BACKGROUND: The aim of the current study was to investigate whether the StethoDop can serve as a valid and reproducible instrument for measuring the ankle-brachial index (ABI) and assessing venous reflux, even when used by inexperienced investigators, in comparison with the classic Doppler. METHODS: I) During four weeks, four ankle-brachial index (ABI) measurements were performed on 44 patients: one measurement with the classic Doppler by an experienced investigator, one with the classic Doppler by an inexperienced investigator and two measurements with the StethoDop by the inexperienced investigator. II) 36 patients were screened for venous insufficiency by detecting venous reflux with the StethoDop and classic Doppler at the saphenofemoral and saphenopoplitial junctions by an inexperienced investigator. The results were compared with the results of the duplex as gold standard and with the results of the examination by an experienced dermatologist with the classic Doppler. RESULTS: I) The confidence interval of ABI measurement for both the classic Doppler and the StethoDop by the inexperienced investigator was within an acceptable +/- 0.21 interval of significant change. II) For venous reflux determination, the overall sensitivity and specificity of the StethoDop were comparable with the sensitivity and specificity of the classic Doppler: sensitivity 76.0 and 75.0%, specificity 94.8 and 94.2%, respectively. The positive predictive value of the StethoDop, compared with the duplex, was 87.5%; the negative predictive value was 90.0%. CONCLUSION: I) For ABI measurement, the StethoDop is a valid instrument with reproducible results, even when used by inexperienced investigators. II) For venous reflux determination, the StethoDop is a valid screening instrument for venous insufficiency. However, as with determination with the classic Doppler, the reflux assessment by StethoDop gives no information about the deep veins and may miss up to 24% of apparent reflux. PMID- 15127832 TI - Bluish-grey pigmentation of fingernails, gingiva, teeth and peri-oral region. Minocycline. PMID- 15127833 TI - Chylous ascites and chylothorax due to constrictive pericarditis in a patient undergoing haemodialysis. AB - Chylous ascites and chylothorax are rare clinical entities and usually caused by neoplasms, particularly lymphomas, liver cirrhosis, superior vena cava thrombosis, nephrotic syndrome, and some cardiac events such as dilated cardiomyopathy or right heart failure. Constrictive pericarditis is an extremely rare cause of this clinical state. We report a 41-year-old male patient undergoing haemodialysis who presented with chylous ascites and chylothorax. Echocardiography and heart catheterisation revealed constrictive pericarditis. He underwent pericardiectomy and after the operation the ascites and pleural effusion resolved rapidly. We suggest that constrictive pericarditis should be considered in the differential diagnosis of chylous ascites and chylothorax. PMID- 15127834 TI - Interstitial pneumonia and hepatitis caused by minocycline. AB - A 28-year-old patient is described who presented with progressive dyspnoea and jaundice due to interstitial pneumonia and hepatitis. The most likely cause is a drug-related reaction to minocycline. We discuss the different kinds of drug related reactions that are most likely involved. PMID- 15127835 TI - Pneumococcal vaccination for healthy elderly: a comment. PMID- 15127836 TI - Antibodies to prion and Acinetobacter peptide sequences in bovine spongiform encephalopathy. AB - An amino acid sequence homology has been identified between the bovine prion sequence (RPVDQ) and the Acinetobacter calcoaceticus enzyme, uridine-diphosphate N-acetyl glucosamine-1-carboxy-vinyl-transferase which also contains (RPVDQ). Class-specific IgA, IgG and IgM antibodies against synthetic peptides containing the structurally related sequences present in bovine prion and A. calcoaceticus were measured in 189 bovine spongiform encephalopathy (BSE) positive cattle, 127 BSE negative cattle and 87 healthy control animals using an ELISA technique. Class-specific IgA, IgG and IgM antibodies against the structurally related synthetic peptides were significantly elevated in BSE positive cattle when compared to BSE negative cattle (P < 0.001) and healthy control animals (P < 0.001). These autoantibodies may have a role in the pathogenesis of BSE. PMID- 15127837 TI - Clinical and immunological effects of Newcastle disease virus vaccine on bovine papillomatosis. AB - Newcastle disease virus (NDV) has antineoplastic and immunostimulatory properties, and it is currently being clinically tested in anticancer therapy. In order to analyze the immunostimulatory effects of NDV on bovine papillomatosis, we inoculated 14 cows subcutaneously with an attenuated vaccine containing the LaSota strain of NDV (LS-NDV). Four cows with papillomatosis served as controls. Serum samples were collected from each animal 1 h before and, 7 and 21 days after inoculation. In inoculated cows, on days 7 and 21 the mean antibody titers were log2 2.43 +/- 0.92 and log2 5.57 +/- 0.72 by haemagglutination inhibition (HI), and the mean levels of tumor necrosis factor-alpha (TNF-alpha) were 5.80 +/- 4.19 and 5.39 +/- 2.66 ng/ml by WEHI-164 cytotoxicity assay. Significant differences between inoculated and control animals were evident for antibody titers on day 21 and clinical scores on day 60. A correlation was evident between the TNF-alpha activities and clinical scores on day 21. The clinical observations at day 60 showed that the papillomas in five cows were completely resolved (36%), one animal had no alterations on clinical appearance of the tumor (7%), and papillomas in eight cows were regressed (57%). In conclusion, these results demonstrated that inoculation of LS-NDV vaccine stimulates an antibody response and limited increase in TNF-alpha activity and may enhance clinical recovery in bovine papillomatosis. PMID- 15127838 TI - In vivo immunostimulatory effects of CpG oligodeoxynucleotide in cattle and sheep. AB - Oligodeoxynucleotides (ODN) containing cytosine-phosphate-guanosine (CpG) motifs have been shown to activate the innate immune system and protect mice and chicken from bacterial and viral infections. Unfortunately, similar studies in other veterinary species are lacking. In this study we assessed the in vivo immunostimulatory effects of CpG ODN 2007, an ODN with previously demonstrated in vitro biological activity. The in vivo effects of ODN 2007 were compared in two closely related outbred species, sheep and cattle, to determine if there were common biological responses. We demonstrated that subcutaneous (s.c.) injection of the CpG ODN induces an acute phase response in the form of a transient fever, a mild transient increase in circulating neutrophils and elevated serum haptoglobin in both sheep and cattle. Sheep injected with CpG ODN also exhibited increased serum 2'5'-oligoadenylate (2'5'-A) synthetase activity, but no increase in serum 2'5'-A synthetase was detected in cattle. The ODN-induced responses were stronger in animals injected with CpG ODN formulated in 30% emulsigen than phosphate buffer saline (PBS) alone. These in vivo data demonstrate for the first time that a CpG ODN induces acute phase immunostimulatory responses in sheep and cattle. However, CpG ODN-induced antiviral effector molecule 2'5'-A synthetase was detected only in sheep but not in cattle. PMID- 15127839 TI - Molecular study on chicken tumor necrosis factor receptor-II and tumor necrosis factor receptor-associated factor-5. AB - Tumor necrosis factor (TNF) receptor-associated factors (TRAFs) were identified as signal transducers for the tumor necrosis factor receptor (TNFR) superfamily. In this study, we cloned and characterized two genes that encode chicken TNFR-II and TRAF5. The initial cDNA fragments were obtained by suppressive subtractive hybridization (SSH) of chicken spleen cells with or without lipopolysaccharide stimulation (Salmonella typhimurium SL1181 (RE-mutant)). The results showed that chicken TNFR-II is 1518 bp in length with an open reading frame (ORF) of 1386 bp having 31% homology with human TNFR-II. Expression analysis of chicken TNFR-II revealed that it is highly expressed in the spleen and bursa of Fabricius. The chicken cell lines IN24, MSB1 and 1104B express TNFR-II abundantly. The time course analysis of expression in spleen, bursa of Fabricius and IN24 cell line showed that TNFR-II is maximally expressed at 6 h after stimulation in bursa of Fabricius and after 8 h stimulation in the IN24 cell line. With regard to TRAF5, the complete sequence was 1936 bp in length with an ORF of 1671 bp that showed 71.3% homology with human TRAF5. Expression analysis showed that, among the tissues examined, TRAF5 was strongly expressed in spleen and bursa of Fabricius, while among the cell lines examined, it was maximally expressed in IN24. Thus, both genes were expressed in the same tissues and cell line among examined materials. These results suggest that chicken TNFR-II may interact with TRAF5 adaptor protein to complete its signal transduction pathway. PMID- 15127840 TI - Recombinant single-chain canine interleukin 12 induces interferon gamma mRNA expression in peripheral blood mononuclear cells of dogs with visceral leishmaniasis. AB - Canine visceral leishmaniasis poses important concerns for public health and veterinary medicine in many areas of the world. Resistance to it seems to be associated with cellular specific immune responses of the so-called Th1 type. Interleukin-12 (IL-12) is one of the most potent inducers of Th1 type of immune responses to co-administered antigens. Herein, the cloning of canine IL-12, as a single-chain fusion protein (sccaIL-12), and its expression in biologically active form in COS-7 cells is reported. Supernatants from these cells stimulated the expression of comparable amounts of interferon gamma mRNA in peripheral blood mononuclear cells from dogs with natural visceral leishmaniasis. In addition, after stimulation with sccaIL-12, there was no difference between interferon gamma mRNA expressions in peripheral blood mononuclear cells of dogs with visceral leishmaniasis and from normal healthy control animals. PMID- 15127841 TI - Protective killed Ehrlichia ruminantium vaccine elicits IFN-gamma responses by CD4+ and CD8+ T lymphocytes in goats. AB - Interferon gamma (IFN-gamma) is considered as a key mediator of protective cell mediated immunity against intracellular pathogens in general, and against Ehrlichia ruminantium, the causative agent of tick-borne heartwater disease of ruminants, in particular. However, the source of this important cytokine in animals immunized against E. ruminantium remains largely unknown. We have analyzed in goats protected by vaccination with a killed E. ruminantium vaccine, the potential of individual, genuine (i.e., non-cloned), T cell subsets to produce IFN-gamma after antigenic recall in vitro. In all vaccinated but none control animals, E. ruminantium-induced IFN-gamma secretion was observed in 24 h stimulated blood. Flow cytometric analysis of stimulated peripheral blood mononuclear cells (PBMCs) collected after each vaccine inoculation indicated that immune CD4+ and CD8+ T cells contribute to the same extent to the production of IFN-gamma, while WC1+ T cells are less important. This was confirmed by blocking the secretion of IFN-gamma with anti-classes I and II major histocompatibility complex antibodies. Blocking experiments also suggest that CD8+ need the help of CD4+ T cells in order to produce IFN-gamma. Thus, this work underlines the key role of CD4+ T cells in the production of IFN-gamma by immune goat PBMC. It also describes, for the first time in ruminants, E. ruminantium-specific CD8+ effector T cells. Since CD4+ and CD8+ T cells collectively contribute to the production of IFN-gamma in most vaccinated animals, and since these responses are associated with protection, it may be that a recombinant vaccine will need to incorporate E. ruminantium antigens capable of driving both responses. PMID- 15127842 TI - Development of a lymphocyte-transformation-assay for peripheral blood lymphocytes of the harbor porpoise and detection of cytokines using the reverse-transcription polymerase chain reaction. AB - Impairment of immune function is suggested to play a contributing role for the increasing incidence of infectious diseases in the harbor porpoise (Phocoena phocoena) of the North and Baltic Seas. Both, lymphocyte-transformation-assay of peripheral blood lymphocytes (PBMC) and detection of cytokine expression are important tools for the characterization of the cellular immune response. To evaluate optimal parameters for the lymphocyte-transformation-assay isolated blood lymphocytes from four healthy harbor porpoises were stimulated with different concentrations of concanavalin A (Con A), phytohemagglutinin (PHA) and pokeweed mitogen (PWM). Cell proliferation was measured photometrically after 72 h using 5-bromo-deoxyuridine-assay and stimulation indices were calculated. The expression of pro- and anti-inflammatory cytokines such as interleukin (IL)-2, IL 4, IL-6, IL-10, transforming growth factor (TGF)-beta, and tumor necrosis factor (TNF)-alpha was investigated in control and mitogen-stimulated lymphocytes using reverse-transcription polymerase chain reaction (RT-PCR). Primers for IL-2, IL-4 and IL-6 were selected from published cDNA-sequences of other cetaceans. Established canine and human primers were taken for the detection of TNF-alpha, TGF-beta, IL-10 and the house keeping transcript glyceraldehyde-3-phosphate dehydrogenase (GAPDH), respectively. Specificity of the amplicon was confirmed by DNA sequence analysis and comparison with nucleotide sequences of other marine and terrestrial mammals. Con A and PHA represented the most powerful mitogens for harbor porpoise lymphoid cells at concentrations of 5 and 2 microg/ml, respectively, while PWM induced a comparatively low maximum proliferation at a concentration of 2 microg/ml. GAPDH was amplified in non-stimulated and all mitogen-stimulated cells. With the exception of IL-10 none of the other cytokines were detected in non-stimulated cells. Transcription of IL-4, IL-6, IL-10, TNF alpha and TGF-beta-mRNA was observed after incubation with all the three phytomitogens, whereas IL-2 was only detected in Con A and PWM treated cells. Lymphocyte-transformation-assay and RT-PCR for detection of cytokines will allow to investigate possible impaired immune function in the harbor porpoise in future studies. PMID- 15127843 TI - Expression of adhesion molecules on milk and blood lymphocytes from periparturient dairy cattle with Johne's disease. AB - Twelve dairy cows infected with Mycobacterium avium subsp. paratuberculosis were monitored for lymphocyte subsets and expression of adhesion molecules on cells in blood and milk at parturition and at intervals up to 21 days post-partum. Using fluorescent antibody labeling of cells and analysis by flow cytometry, we determined percentages of T cell subsets (CD4+, CD8+, gammadelta+) and expression of adhesion molecules (CD62L, LFA-1, LPAM-1, and CD44) on cells from blood and milk of these cows. Significantly higher percentages of CD8+ cells were found in milk than in blood at all time points; there were no significant differences in percentages of CD4+ or gammadelta+ cells. CD62L, LFA-1, and LPAM-1 were expressed on a significantly higher percentage of all T cell subsets in milk than in blood at various times after parturition. No differences were seen in expression of CD44. Increased percentages of T lymphocytes expressing adhesion molecules in milk compared to blood suggest that a migratory population of cells is being selectively recruited to the mammary gland from the circulation. PMID- 15127845 TI - The neonatal Fc receptor (FcRn) is expressed in the bovine lung. AB - In neonatal calves, maternal immunoglobulin (Ig) is transferred into respiratory secretion which contributes to protection against pathogens. The early predominance of IgG1 in respiratory tract secretions is progressively reduced in favor of IgA by age but in the lower, bronchoalveolar system secreted IgG remains the dominant secreted Ig even in adulthood. The trans-epithelial transport of secretory IgA into mucosal secretions is carried out by the polymeric Ig receptor. However, the mechanism by which IgG crosses epithelial cells to provide defense on mucosal surfaces is still unknown. In order to investigate the possibility that the neonatal Fc receptor, FcRn is involved in this transport we have first analyzed the localization of this receptor in the upper and lower respiratory tracts. Consistent with the in situ hybridization data, immunohistochemistry showed undetectable expression in the tracheal epithelial cells, relatively weak expression in epithelial cells of the bronchi, apparent staining those lining the bronchioli and randomly scattered signal over the alveolar tissue. The bovine FcRn may thus play a role in IgG transport across mucosal epithelial barriers as a trafficking receptor and ensure IgG predominance in the lower respiratory tract. PMID- 15127844 TI - Antibody isotype profiles in serum and circulating antibody-secreting cells following mucosal and peripheral immunisations of sheep. AB - The isotype-specific antibody responses of sheep immunised with keyhole limpet hemocyanin by a peripheral route (intramuscular (i.m.) injection) were compared to those induced by immunisation via different mucosal routes: (1) intra-nasal spray; (2) rectal deposition with cholera toxin; (3) injection into the mucosa of the small intestine or rectum. Antigen-specific IgG1 antibodies were induced in the i.m., intra-intestinal and intra-rectal injection groups and in a proportion of the cholera toxin immunised sheep, but not in the intra-nasal immunisation group. IgA was the only antibody isotype detected in serum collected from the intra-nasal immunisation group. No significant differences in serum IgA levels were detected in any of the mucosal immunisation groups as compared to the i.m. injection group. In contrast, analysis of the in vitro antibody profiles secreted by circulating antibody-secreting cells (ASC) revealed significantly higher IgA responses in the supernatants from all mucosal immunisation groups. This suggests that the measurement of antibodies secreted by circulating ASCs may be a better correlate of local mucosal responses in ruminants, as has been previously demonstrated in human studies. In addition to IgG1 and IgA responses, immunisation by direct injection of antigen formulations into the intestinal and rectal mucosa were the only groups to induce consistently high IgG2 antibodies in serum and ASC cultures. PMID- 15127847 TI - Failure of FIV-infected cats to control Toxoplasma gondii correlates with reduced IL2, IL6, and IL12 and elevated IL10 expression by lymph node T cells. AB - Increased susceptibility to intracellular pathogens in HIV-infected individuals and FIV-infected cats is attributed to a defective T-helper 1 (Th1) immune response. However, little is known about specific cytokine responses to secondary pathogens. To address this question, control and FIV-infected cats were challenged with Toxoplasma gondii, and lymph node cells analyzed for cytokine mRNA expression. Twenty-four weeks post-FIV infection, prior to T. gondii challenge, IL2 and IL12 mRNAs were depressed, whereas IL10 and IFNgamma mRNAs were increased in CD4+ and CD8+ subsets. Following T. gondii challenge, control cats showed increased expression of IL2, IFNgamma, IL10, IL12, and IL6 mRNAs. In contrast, IL2, IL6, IFNgamma, and IL12 mRNAs were suppressed in FIV-T. gondii co infected cats, whereas IL10 remained at the high prechallenge levels. IFNgamma and IL10 mRNAs were produced by both CD4+ and CD8+ cells in FIV-T. gondii cats. Elevated IL10 may suppress a Th1 cytokine response to T. gondii challenge. PMID- 15127846 TI - The immunogenicity of Rhodococcus equi GroEL2-based vaccines in a murine model. AB - Rhodococcus equi is a significant intracellular bacterial pathogen in foals. However, at present there is no commercially available vaccine for the prevention of R. equi-induced disease in these animals. Studies have shown that GroEL based vaccines can afford protection against some intracellular pathogens. In this study, the R. equi gene encoding the heat shock protein GroEL2 was cloned and sequenced, with a view to using it as a vaccine candidate. The promoter region of the gene contained two copies of controlling inverted repeat of chaperone expression (CIRCE) motifs, which are well-recognised transcriptional regulators of bacterial heat shock proteins. The R. equi GroEL2 was expressed in E. coli BL21 DE3 with a C-terminal His-tag and sequenced to confirm its identity. The R. equi purified His-tagged GroEL2 protein and a groEL2-based DNA vaccine were used in separate experiments to immunise BALB/c mice. The recombinant protein-based vaccine elicited a mixed Th1/Th2 response whereas the DNA vaccine was found to elicit a predominantly Th1 biased immune response. However, when vaccinated mice were challenged intravenously with 1.5 x 10(7) R. equi neither vaccine elicited enhanced bacterial clearance from the spleen or liver in this model. The reasons for this apparent lack of success are discussed. PMID- 15127848 TI - The status of vascular surgery as an independent specialty in Europe: are the relationships with general and cardiothoracic surgery a problem? AB - Vascular surgery has established a clear clinical and scientific profile in Europe over the last decade, but it presents a highly complex, disorganized, and unplanned pattern. It is a specialty in the majority of the present member states of the European Union (EU), but in the United Kingdom, Germany, the Netherlands, and Sweden this is not the case. With the current expansion of the EU with 13 countries, mainly from the former Eastern Bloc, it will be even more necessary to ensure at least some level of convergence in the standards of training, certification, quality assured practice, continuing medical education, recertification, access to and quality of care, etc, because free migration of doctors, and patients, is a derivative of the cornerstones of the EU treaties, namely free movement of the citizens (and capital). The profession has been successful in creating a simple and coherent system for organization within the European Union of Medical Specialists with a board of vascular surgery, for a (voluntary) European proficiency test for specialists (the EBSQ-Vasc), and for European continuing medical education, all in close collaboration with the premier scientific society, the European Society for Vascular Surgery. The fantastic reductions in working hours for young doctors in the EU represent a serious threat to standards of training and, ultimately, to the patients. This, in connection with increased litigation and compensation demands for incompetence and negligence, makes it even more necessary to establish European minimum standards for training, professional competence, and an obligatory European specialist examination. A key element in this difficult process is the establishment of vascular surgery as a specialty in all member states. Day-to-day collaboration with radiologic interventionalists has developed pragmatically at the local level in most places, but only when the administrative structures mature and vascular surgery becomes a specialty in all countries will the necessary tools be available for the avoidance of professional conflicts with colleagues from other specialties. PMID- 15127849 TI - What changes are needed to keep vascular surgery alive and vigorous? AB - To keep vascular surgery alive and vigorous, we need to regain our preeminence as the principal therapists for patients with aneurysms, carotids, and occlusive disease of the lower extremities. We must regain the sense of excitement about our specialty and refocus training for residents and practicing surgeons toward those skills necessary to provide the full range of therapies at the highest level. Attaining these goals will require embracing, obtaining, and applying endovascular expertise throughout the vascular workforce. We can no longer moan and whine over encroachments into our turf by nonsurgeons. We must make our own opportunities. Changes in our attitude, our identity, and our structure will be necessary to do so. Our ability to make change will depend upon our values, defined as the judgment as to whether we remain a derivative of our general surgical roots (with continued emphasis on improved quality from traditional operations) or embrace the disruptive technology of endoluminal therapy as the next iteration of vascular surgical practice. We cannot do both because the preparation for the latter precludes the former. We are at a crossroads because if we accept endovascular technologies as critical to our future, our path is clear. PMID- 15127850 TI - American board of vascular surgery: the first 7 years. AB - The American Board of Vascular Surgery (ABVS) was incorporated in 1996 with a vision of improved training standards and certification of vascular surgeons. At that time, 91% of those holding American Board of Surgery Certificates of Added Qualifications in Vascular Surgery supported the formation of the ABVS. Subsequent events have led to a clear definition of specific educational issues important to the vascular surgery community. Unresolved issues relate to the need to complete a general surgery residency before beginning a vascular surgery fellowship, the continued inclusion of vascular surgery as a primary component of general surgery training, and the absence of a designated Residency Review Committee for Vascular Surgery. These issues have persisted since the inception of the ABVS. An application for the ABVS to become an American Board of Medical Specialties (ABMS) primary board was submitted in 2002 with a preliminary hearing before a liaison committee composed of American Medical Association and ABMS members. The American Board of Surgery (ABS) and a minority of the vascular surgery community vigorously opposed the application. The perceived divisiveness created by their actions contributed to the application's initial rejection and the necessity for an appeal. Certain ABS directors have recently stated that they would consider approving multiple track-type training that could allow single certification in vascular surgery, following 5 to 6 years of postgraduate training after medical school. The ABVS cautiously supports this action, recognizing that this radical change for the ABS may not be feasible given the broad-ranging interests of general surgery and restrictive ABMS guidelines for certifying medical specialists. The impact of not resolving the critical issues facing vascular surgery in a timely manner is that there will be inadequate numbers of competent vascular surgeons to provide for society's needs. An independent ABMS-approved ABVS provides a clear opportunity to resolve the recognized failings of the status quo. PMID- 15127851 TI - Who in vascular surgery will have time for research today and in the future? PMID- 15127852 TI - An update on the important findings from the EUROSTAR EVAR registry. AB - This article summarizes the important findings to emerge from the European Collaborators on Stent-Graft Techniques for Abdominal Aortic Aneurysm Repair (EUROSTAR) Registry since its inception in 1996. The Registry is an open observational study of the outcomes from endovascular aneurysm repair (EVAR). Data have been contributed, on a voluntary basis, by 135 vascular centers from 18 European countries. Patients are registered prospectively, and data analysis is on an intention-to-treat basis. Results from the first generations of endograft were characterized by excellent early results but poor durability of abdominal aortic aneurysm repair. EUROSTAR data have played a vital role in informing the technical evolution of subsequent generations of endograft and their clinical application. The latest data from EUROSTAR show significant improvements in all outcome measures, most importantly, those relating to durability. EUROSTAR has tracked and informed the evolution of EVAR. Improving results confirm that substantial progress has been made since 1996. EUROSTAR continues to contribute to this ongoing process. PMID- 15127853 TI - Problems facing vascular surgery in 2004. AB - The most pressing challenges we face in the immediate future are endovascular training for those already in practice and a new educational paradigm for our residents. A number of avenues for training those in practice have been implemented, and newer methods, including computer simulation, are being explored. Vascular training programs should be 3 years in length, include vascular navigation and interventional skills, and follow 3 years of basic general surgical training. These changes in our resident training paradigm can take place only through two avenues: a successful reapplication to the American Board of Medical Specialties for an independent American Board of Vascular Surgery or a full reconfiguration of the training programs for general and vascular surgeons that would have to be spearheaded by the American Board of Surgery. There is skepticism that the latter could take place in the limited time we have left to make the vascular surgery residency attractive to candidates and sufficient in experiential and knowledge content. PMID- 15127854 TI - Technical note on renal angioplasty and stenting: new developments that facilitate its performance. AB - Technologic advances in percutaneous catheter-based therapy for renovascular occlusive disease have resulted in progressively miniaturized wires, balloon catheters, stents, and delivery systems. The technique of angioplasty and stenting of renal artery stenoses has been shown to be a safe and effective option for severe hypertension and ischemic nephropathy. Renal artery stenting, especially when performed with the newer lower-profile systems, can be performed with minimal morbidity and a reliably high degree of initial technical success. In this article, we describe our technique using a lower-profile platform of balloons and stents and review data supporting renal artery stenting. PMID- 15127855 TI - Should older patients be selectively referred to high-volume centers for abdominal aortic surgery? AB - The objective of the current study was to determine the effect of hospital volume on outcomes of abdominal aortic surgery for patients older than and younger than 65 years. In order to perform this investigation, information on all adult patients who underwent abdominal aortic surgery in Maryland from 1994 to 1996 (N = 2,987 patients) in 45 acute care hospitals was obtained. Hospitals were designated as low (< 20/year), medium (20 to 36/year), or high (> 36/year) volume according to the annual number of procedures performed. The relationship of hospital volume and mortality was determined for patients less than or greater than 65 years old. Two separate multiple logistic regression models were used to adjust for patient case-mix in each age category. Of the 2,987 patients, 2,067 (69%) were older than 65 years and 920 (31%) were younger. The crude in-hospital mortality rates according to hospital volume were 2.7% (low), 2.1% (medium), and 2.7% (high) for patients younger than 65 years old (p = .8). For patients older than 65 years, in-hospital mortality rates were 11.9% (low), 9.9% (medium), and 6.9% (high) (p = .005). After adjusting for patient case-mix in a multivariate analysis, high hospital volume was associated with a decreased risk of in hospital mortality for patients older than 65 years (OR 0.57; 95% CI 0.37 to 0.86; p = .008) but not for patients under 65 years old. In conclusion, hospital volume was associated with decreased in-hospital mortality after abdominal aortic surgery only for patients greater than 65 years old. Because of this differential effect, targeting elderly patients for regionalization would achieve most potentially avoidable deaths for this common high-risk surgical procedure. PMID- 15127856 TI - Arterial reconstruction in the upper extremities. AB - Arterial reconstruction in the upper extremities is rare relative to the incidence of reconstruction in the lower extremities. Twenty-three patients who underwent vascular reconstruction in the upper extremities from 1985 to 2000 were retrospectively reviewed. Atherosclerosis was observed in only two subclavian arteries. The most common cause of occlusion was thromboembolism from the heart, which occurred often in the brachial artery and could be treated successfully with thrombectomy. Seven of eight bypass grafts, including three procedures below the elbow, remained patent during follow-up periods ranging from 2 to 9 years. Arterial reconstruction of the upper extremities differs from that in the lower extremities. Atherosclerosis is a rare cause of ischemia, and the etiology varies with the site of obstruction. Bypass surgery below the elbow is feasible and successful in selected patients. Ischemia of the upper extremity causes significant morbidity and should be treated aggressively whenever possible. PMID- 15127857 TI - What have we learned from the Imaging in Carotid Angioplasty and Risk of Stroke (ICAROS) study? AB - Carotid artery stenting (CAS) is used widely to treat carotid lesions. Indication to CAS is mostly based, both in trial and in clinical practice, on the percentage of stenosis and the presence or absence of preprocedural neurologic symptoms, whereas the features of the plaque are somehow disregarded and ignored. The most severe complication of CAS is stroke, related to cerebral embolization from carotid plaque. Several studies showed that echolucent plaques generate a higher number of embolic particles following carotid stenting. Echolucency can be measured using the gray scale median, which is an objective and quantitative computer-assisted grading of the echogenicity of carotid plaques. As previously demonstrated in the ICAROS study, carotid plaque echolucency is an independent risk factor for stroke in carotid stenting. Carotid plaque echolucency is one of the parameters that should be mandatory to be considered for indication to treatment. PMID- 15127858 TI - Effect of graduated compression stockings on limb oxygenation and venous function during exercise in patients with venous insufficiency. AB - Despite the established role of compression as the basis for nonoperative treatment of chronic venous insufficiency (CVI), its mechanism of action remains unclear. Near-infrared spectroscopy (NIRS) provides continuous noninvasive monitoring of changes in tissue oxyhemoglobin (HbO2) and deoxyhemoglobin (Hb). We applied NIRS to evaluate the effect of graded stockings on venous function and calf muscle oxygenation during exercise in patients with CVI. Ten patients (age 56 +/- 5 years) with CVI were rested in supine posture for 20 minutes. NIRS optodes were attached to the calf. Venous function was assessed in each patient with and without graded compression stockings (classes I to III) at rest in the supine position, standing, with 10 tiptoe exercises, and on standard walking at 1.6 km/h for 5 minutes. Venous function was assessed by measuring changes in Hb and total hemoglobin (HbT) during the test, and muscle oxygenation was assessed by the oxygenation index (HbD), which is the difference between HbO2 and Hb. Standing without stockings caused a significant increase in Hb concentration by 10.75 +/- 2.24 micromol/L compared with the supine position (p < .001). This value was reduced when stockings were applied to 6.38 +/- 2.75 micromol/L with class III stockings (p = .005). During tiptoe exercise, the residual Hb concentration value without stockings was 7.62 +/- 2.12 micromol/L compared with 5.88 +/- 2.87, 3.77 +/- 3.37, and 3.46 +/- 2.73 micromol/L for class I, II, and III stockings, respectively. The reduction in Hb concentration reached significance with class II and III stockings compared to without stockings (p = .04). The HbT concentration was also reduced during tiptoe exercise, with increasing compression from 15.46 +/- 5.31 micromol/L without compression to 11.52 +/- 4.26 pmol/L with class III stockings (p = .048). During walking, the Hb concentration was 11.40 +/- 3.10 pmol/L without stockings, decreasing significantly (p < .001) and progressively to 8.49 +/- 3.24, 7.71 +/- 3.51, and 6.89 +/- 3.16 micromol/L with class I, II, and III stockings, respectively. Limb oxygenation (as measured by HbO2 concentration) during walking exercise, however, increased with higher-compression stockings and reached significance with class III stockings only (p = .03). In patients with venous insufficiency, graduated compression stockings may achieve their beneficial effects by reducing venous pooling and improving deeper tissue oxygenation. PMID- 15127859 TI - A perfect storm: lessons from the vaccine controversy. PMID- 15127860 TI - Stress and periodontitis. PMID- 15127861 TI - Nitrous oxide. PMID- 15127862 TI - Total-etch versus self-etch. PMID- 15127863 TI - Do you sometimes provide dental care to disadvantaged patients who may not be able to pay for your services? PMID- 15127864 TI - Aerosols and splatter in dentistry: a brief review of the literature and infection control implications. AB - BACKGROUND: Aerosols and droplets are produced during many dental procedures. With the advent of the droplet-spread disease severe acute respiratory syndrome, or SARS, a review of the infection control procedures for aerosols is warranted. TYPES OF STUDIES REVIEWED: The authors reviewed representative medical and dental literature for studies and reports that documented the spread of disease through an airborne route. They also reviewed the dental literature for representative studies of contamination from various dental procedures and methods of reducing airborne contamination from those procedures. RESULTS: The airborne spread of measles, tuberculosis and SARS is well-documented in the medical literature. The dental literature shows that many dental procedures produce aerosols and droplets that are contaminated with bacteria and blood. These aerosols represent a potential route for disease transmission. The literature also documents that airborne contamination can be minimized easily and inexpensively by layering several infection control steps into the routine precautions used during all dental procedures. CLINICAL IMPLICATIONS: In addition to the routine use of standard barriers such as masks and gloves, the universal use of preprocedural rinses and high-volume evacuation is recommended. PMID- 15127865 TI - Should dentists become 'oral physicians'? Yes, dentists should become 'oral physicians'. AB - In summary, a new designation for the dental profession is warranted, not because paramedical professionals have done it, but because the new designation is appropriate for what they are able to do. Moreover, it can be done now with little resistance or fanfare. As has been suggested by others, changing the name "dentist" to the name "oral physician" would result in several benefits: dentists' being recognized as providers of services such as tobacco-use cessation, oral cancer screenings, nutritional counseling and, most recently, as a major health care resource for dealing with bioterrorism; the public's visiting dental professionals for services other than traditional dental procedures; the profession's being more likely to teach and provide services outside of traditional dental procedures; third-party payers' being more likely to pay for services other than traditional dental procedures; the improvement in the public's oral health that would result from patients' visiting "oral physicians" for services other than traditional dental procedures. PMID- 15127866 TI - Should dentists become 'oral physicians'? No, dentistry must remain dentistry. AB - Dentistry is not an allied health profession. It is not a paramedical profession. It is time that dentistry be recognized as the profession that offers patients some of the most complex surgery performed on the human body--namely, restorative dentistry and rehabilitation of the masticatory system. Dentistry is the only anatomically focused health care profession that is university-based and for which primary care responsibility is maintained by the profession. An inferiority complex about what it means to be a dentist has served only to confuse the public and bring us further from our goal of improving the health of all our patients. This inferiority complex is driven by the public and the medical profession, neither of which understands how dentistry fits into overall health care. It is essential that every academic health center have oral health education as an integrated part of health care education for dentists, physicians, nurses, allied dental personel, physical therapists, psychologists and all who receive university-based health care education. In this way, all the health professions and the public will see dentistry and oral health as essential to patients' overall health. The idea of emulating those who do not have the strength of basic science education, practice complexity, surgical skills or community status by seizing a new title will not elevate the profession for the future. The public knows what a dentist is. It is our task to inform the public about the capabilities of dentists and the value of oral health and our profession. We can accomplish this best by assuring that our profession's name, "dentistry," is understood to represent one of the world's most accomplished surgical endeavors, one that is thoroughly integrated into the fabric of health care. Thus, good oral health will be thoroughly integrated into what it means to be healthy. PMID- 15127868 TI - Provisionally restoring a necrotic tooth while maintaining root canal access. PMID- 15127867 TI - A three-year clinical evaluation of two dentin bonding agents. AB - BACKGROUND: A new restorative called a "giomer composite" has been introduced. The authors conducted a study to determine retention, anatomical form, caries, staining, marginal discoloration, marginal adaptation, surface roughness and sensitivity of giomer compared with those of a microfilled composite. METHODS: The authors placed 40 sets of restorations randomly in canines and premolars in vivo. They used a giomer composite and a microfilled composite in erosion/abrasion/abfraction Class V lesions that were not altered with rotary instruments. They placed the restorations according to manufacturer's recommendations, and two calibrated examiners evaluated the restorations independently using modified U.S. Public Health Service criteria at baseline and at six, 18 and 36 months. The lesions receiving the restorations did not differ from each other in the amount of circumferential enamel present, the percentage of the surface area of dentin or lesion type. RESULTS: There were no differences in the restorations at baseline, an evaluation made two weeks after placement. At 36 months, the giomer and microfilled composite restorations were not significantly different from one another in any of the eight criteria evaluated. The percentage agreement between examiners was at least 83 percent for each criterion in each evaluation period. CONCLUSIONS: Both the giomer and the microfilled composite used in this study meet the clinical portion of the Acceptance Program Guidelines for Dentin and Enamel Adhesives Materials established by the American Dental Association. CLINICAL IMPLICATIONS: Both the giomer and the microfilled composite used in this study can be used with confidence in Class V lesions. PMID- 15127869 TI - The light-curing mania. AB - In the approximately 25 years that dental restorative resins have been cured by light, there has been constant evolution of and improvement in the curing devices and the resins they cure. During the past decade, curing lights have become very effective. Dentists who have functional halogen, fast halogen or PAC lights may wish to compare their current lights with the new generation of LED lights that are dominating the commercial dental advertising at this time. If the new LED lights offer more than the current light they are using, they might consider changing to an LED. If their current light offers about the same characteristics as the LED light being considered, I do not recommend changing. I anticipate that continued upgrading and improvement in LED lights eventually will make this concept the most used method for curing restorative resin. PMID- 15127870 TI - Survey of systematic reviews in dentistry. AB - BACKGROUND: Although systematic reviews are the backbone of evidence-based dentistry, they have appeared infrequently in the clinical dental literature and their importance may not be recognized by dentists. The authors describe the steps taken in systematic reviews and perform a literature survey to identify published systematic reviews of topics relevant to clinical dentistry. METHODS: The authors searched MEDLINE and the Cochrane databases of systematic reviews and abstracts of reviews of effectiveness, as well as identified reviews that were known to the authors but not found in the searches. Systematic reviews included in this survey stated the intention to identify all relevant articles within predefined limitations, applied defined exclusion and inclusion criteria, and presented complete raw or synthesized data from included studies. RESULTS: This literature survey identified 131 systematic reviews, 96 of which had direct clinical relevance. During the past 14 years, clinically relevant systematic reviews have been published with increasing frequency. These reviews vary in the types of studies included and the assessment of those studies. The results of the reviews also varied in their definitiveness, with 17 percent finding the evidence to be insufficient to answer the key question. An additional 50 percent of the 96 reviews hedged in answering the key question, by noting that the supporting evidence was weak in quality or limited in quantity. CONCLUSION: The number of systematic reviews that address clinical topics in dentistry is small but growing. However, the authors of more than one-half of these reviews believed that the evidence available to answer the key question was not strong. CLINICAL IMPLICATIONS: As systematic reviews continue to grow, dentistry will become better informed about the adequacy and congruence of the scientific evidence underpinning clinical practice. PMID- 15127871 TI - Biopsychosocial differences between high-risk and low-risk patients with acute TMD-related pain. AB - BACKGROUND: The aim of this study was to use a biopsychosocial perspective to characterize patients who were identified as being at high risk, or HR, of progressing from acute to chronic jaw-related pain. METHODS: The authors classified 74 subjects as being at HR or low risk, or LR, according to the predictive algorithm. They used a variety of functional and biopsychosocial measures to evaluate subjects. RESULTS: The HR group had significantly higher levels of self-reported pain as measured by the Characteristic Pain Inventory and significantly higher levels of depression as measured by the Beck Depression Inventory-II. They were 11 times more likely to have a Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, or DSM-IV, Axis I clinical diagnosis, and more than three times as likely to have a DSM-IV Axis II personality disorder. Logistic regression analyses identified variables that differentiated, with 77 percent accuracy, the HR and LR patients. CONCLUSIONS: Overall, the HR subjects had more psychopathology than did the LR subjects, used poorer coping styles and had greater self-reported pain. Six psychosocial factors alone enabled the authors to correctly classify 77 percent of the subjects as being in the HR group. CLINICAL IMPLICATIONS: Future research, in conjunction with the above findings, may enable the authors to determine, with greater certainty, if patients who are more anxious are at greater risk of developing chronic pain. If so, this provides further evidence of the need for early detection of patients at risk of developing chronic pain and the need to refer them for adjunctive care, such as cognitive-behavioral intervention. PMID- 15127872 TI - Combating antibiotic resistance. AB - BACKGROUND: The ADA Council on Scientific Affairs developed this report to provide dental professionals with current information on antibiotic resistance and related considerations about the clinical use of antibiotics that are unique to the practice of dentistry. OVERVIEW: This report addresses the association between the overuse of antibiotics and the development of resistant bacteria. The Council also presents a set of clinical guidelines that urges dentists to consider using narrow-spectrum antibacterial drugs in simple infections to minimize disturbance of the normal microflora, and to preserve the use of broad spectrum drugs for more complex infections. CONCLUSIONS AND PRACTICE IMPLICATIONS: The Council recommends the prudent and appropriate use of antibacterial drugs to prolong their efficacy and promotes reserving their use for the management of active infectious disease and the prevention of hematogenously spread infection, such as infective endocarditis or total joint infection, in high-risk patients. PMID- 15127873 TI - Managing time. AB - As you can see from these examples, there are many factors that must be considered in the relationship between time management and practice production. However, it is important to understand that there is a direct correlation among various factors--including how time is used, which services are provided, the volume of services provided and other factors--in establishing a practice with excellent financial results. As always, dentistry is about excellent patient care, but it still is important for the dentist and team to examine the day-to day use of time and its effect on the business side of the practice. PMID- 15127874 TI - Access to dental care in Alabama for children with special needs: parents' perspectives. AB - BACKGROUND: Access to dental care and delivery of quality dental health services are important for children with special needs. The authors surveyed parents of children with special needs in Alabama to determine their perceptions of access and barriers to dental care for their children. METHODS: The authors sent a questionnaire to 2,057 parents of children aged 3 to 13 years with special needs- cleft lip and/or palate; cerebral palsy, or CP; spina bifida; or epilepsy/seizure disorders--who were listed in a database provided by Children Rehabilitation Services of Alabama. The authors conducted univariate and multivariate analyses to calculate odds ratios and 95 percent confidence intervals. RESULTS: The overall response rate was 38 percent (N = 714). Eighty-five percent of respondents reported that their children had received some form of routine dental care. However, 35 percent of respondents reported they had had problems finding dentists willing to treat their children. Among those with problems, significant barriers to dental care included their children's having Medicaid insurance, poor oral health or CP, as well as a shortage of dentists with training in the care of children with special needs. CONCLUSIONS AND PRACTICE IMPLICATIONS: While the majority of respondents said their children had access to dental care, one-third said their children had problems receiving this care. Many of these problems can be ameliorated. Increasing providers' participation in the Medicaid program and improving their knowledge about, empathy for and training in the care of children with special needs is essential in improving access to dental care for this population. PMID- 15127876 TI - Reverse age discrimination? Supreme Court says law does not bar discrimination against younger workers. PMID- 15127877 TI - For the dental patient. Careers in dentistry: are you interested in becoming a dental assistant? PMID- 15127875 TI - Adjunctive benefit of an essential oil-containing mouthrinse in reducing plaque and gingivitis in patients who brush and floss regularly: a six-month study. AB - BACKGROUND: Mechanical methods of oral hygiene can be complemented by the use of chemotherapeutic mouthrinses. The authors sought to quantify the additional benefit provided by an essential oil-, or EO-, containing mouthrinse in reducing plaque and gingivitis in patients who brush and floss regularly. METHODS: The authors randomly assigned patients with gingivitis to one of three treatment groups: brushing and rinsing with a control mouthrinse, or BC; brushing, flossing and rinsing with a control mouthrinse, or BFC; or brushing, flossing and rinsing with an EO-containing mouthrinse, or BFEO. Patients received a dental prophylaxis at baseline, and the authors followed them for six months. RESULTS: Of 246 enrolled subjects enrolled in the study, 237 subjects were evaluable at the study's conclusion. After six months, the subjects using the BFEO regimen had statistically and clinically significant lower mean Modified Gingival Index, or MGI, scores and Plaque Index, or PI, scores than did subjects in the BC group (29.9 percent and 56.3 percent, respectively; P < .001). Subjects in the BFC group had statistically significantly lower mean MGI and PI scores than did subjects in the BC group (11.2 percent and 9.3 percent, respectively; P < .001). Subjects in the BFEO group exhibited statistically and clinically significantly lower mean scores for MGI and PI than did subjects in the BFC group (21 percent and 51.9 percent, respectively; P < .001). CONCLUSIONS: This study confirms that for patients with gingivitis who brush and floss routinely, the adjunctive use of an EO-containing mouthrinse provides a clinically significant and meaningful additional benefit in reducing plaque and gingivitis. CLINICAL IMPLICATIONS: An EO-containing mouthrinse is an effective adjunct to regular brushing and flossing. Therefore, the BFEO regimen can be beneficial for patients with gingival inflammation. PMID- 15127878 TI - Renal vascular resistance and renin-angiotensin system in the pathogenesis of early hypertension in autosomal dominant polycystic kidney disease. AB - Activation of the renin-angiotensin system (RAS) has been proposed to increase renal vascular resistance (RVR) and to play a role in the development of hypertension in autosomal dominant polycystic kidney disease (ADPKD). The aim of this study was to investigate the relationship among RVR, RAS and blood pressure (BP) profile in patients without renal impairment. Thirty-four ADPKD patients underwent ambulatory blood pressure monitoring (ABPM) over a 24-h period and were divided into two groups: 17 hypertensive (group A, day-systolic BP > or = 135 mmHg and/or day-diastolic BP > or = 85 mmHg) and 17 normotensive (group B, day-BP < 135/85 mmHg) patients. The two groups were comparable with respect to age, sex, and renal function. None of the patients assumed therapy. In all subjects the plasma renin activity (PRA) was measured, and the RVR was assessed by measuring resistivity indices (RI). RI was significantly higher in the hypertensive than in normotensive patients (0.67 +/- 0.05 vs. 0.62 +/- 0.03), while PRA was normal in all subjects, and showed no statistical difference between the two groups. Taking all the patients together (group A + group B), a significant positive correlation between RI and 24-h mean arterial pressure (MAP) was discovered, but no correlation was found between RI and PRA or between MAP and PRA. We conclude that in ADPKD patients without renal impairment the MAP values are strictly correlated with the RVR, but not with PRA. Thus factors other than RAS probably contribute to the increase of the RVR and to the early development of hypertension. PMID- 15127879 TI - Hyperuricemia and cardiovascular risk factor clustering in a screened cohort in Okinawa, Japan. AB - The relation between serum uric acid level and cardiovascular risk factors is complex and has been investigated mainly in men. We examined the correlation between serum uric acid level and obesity, hypertension, dyslipidemia, and diabetes mellitus (DM) in both men and women of a screened cohort in Okinawa, Japan. A total of 9,914 individuals (6,163 men and 3,751 women ranging in age from 18 to 89 years) who were screened at Okinawa General Health Maintenance Association were subjects in this study. Hyperuricemia was defined as a serum uric acid level > or = 7.0 mg/dl in men and > or = 6.0 mg/dl in women. The odds ratios (95% confidence intervals) for the presence of hyperuricemia in men were 1.75 (1.56-1.97) for obesity, 1.42 (1.25-1.62) for hypertension, 1.16 (1.02-1.30) for hypercholesterolemia, 1.80 (1.60-2.03) for hypertriglyceridemia, 1.19 (1.02 1.40) for hypo-high density lipoprotein (HDL) cholesterolemia, and 0.61 (0.49 0.75) for DM; in women, they were 2.02 (1.62-2.53) for obesity, 1.64 (1.29-2.10) for hypertension, 1.31 (1.04-1.65) for hypercholesterolemia, 1.95 (1.51-2.51) for hypertriglyceridemia, 1.53 (0.96-2.44) for hypo-HDL cholesterolemia, and 1.20 (0.76-1.90) for DM. Hyperuricemic subjects had higher rates of coexistence of two or more of these cardiovascular risk factors than non-hyperuricemic subjects (63.8% vs. 43.2% in men; 58.9% vs. 27.6% in women). The present study revealed that hyperuricemia is positively associated with obesity, hypertension, and dyslipidemia in both men and women, and that hyperuricemic subjects tend to have a clustering of these cardiovascular risk factors. PMID- 15127881 TI - Relationship between the awareness of salt restriction and the actual salt intake in hypertensive patients. AB - A 24-h home urine collection was conducted to estimate accurate salt intake in hypertensive outpatients. Using 24-h urinary creatinine excretion as a criterion for success, urine samples were obtained from 534 hypertensive patients. The urinary salt excretion of hypertensive outpatients ranged widely from 1.5 to 23.4 g/day (mean value 9.7 +/- 3.9 g/day). Urinary salt excretion was higher in males than in females (10.6 +/- 4.0 vs. 9.2 +/- 3.7 g/day, p<0.01). Based on the questionnaires, the patients were divided into salt-conscious patients, or those who were careful to reduce their daily salt intake, and non-salt-conscious patients. It was found that urinary salt excretion was lower in the salt conscious group than in the non-salt-conscious group (9.4 +/- 3.8 vs. 10.6 +/- 4.0 g/day, p<0.01), but that urinary salt excretion adjusted for body weight was not significantly different between the two groups (0.16 +/- 0.06 vs. 0.17 +/- 0.07 g/kg/day). Our results suggest that there was no obvious reduction in the actual salt intake in salt-conscious patients, suggesting the importance of monitoring salt intake by 24-h home urine collection and informing patients of their actual salt intake as a means of encouraging the achievement of salt restriction. PMID- 15127880 TI - Greater change of orthostatic blood pressure is related to silent cerebral infarct and cardiac overload in hypertensive subjects. AB - Greater change of postural blood pressure (BP) is often seen in elderly hypertensives and is recognized as a risk factor for cognitive decline and poorer cerebrovascular outcome, but its clinical significance still remains to be clarified. We performed a head-up tilting test, ambulatory BP monitoring, and brain MRI in 59 hypertensives and 27 normotensive subjects. We measured plasma atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) levels at rest to assess cardiac burden. The 59 hypertensive patients were classified into 3 groups: an orthostatic hypertension (OHT) group with orthostatic increase in systolic BP (SBP) > or = 10 mmHg (n=16); an orthostatic hypotension (OHYPO) group with orthostatic SBP decrease < or = -10 mmHg (n=18); and an orthostatic normotension (ONT) group with neither of these two patterns (n=25). A group of 27 normotensive subjects (NT) was also included as a control. Plasma BNP (72 +/- 92 vs. 29 +/- 24 pg/ml, p < 0.05) and BNP/ANP ratio (4.6 +/- 3.3 vs. 2.4 +/- 1.5, p < 0.05) were significantly higher in the OHYPO than in the NT group. The BNP/ANP ratio was also higher in the OHT than in the NT group (5.1 +/- 3.9 vs. 2.4 +/- 1.5, p < 0.01). The number of silent cerebral infarct (SCI), prevalence of SCI and number of multiple SCIs was the highest in the OHT group, followed in order by the OHYPO, ONT and NT groups. Blood pressure and left ventricular mass index were not significantly different among the 3 hypertensive groups. In conclusion, hypertensive patients with greater change of postural BP (OHT and OHYPO) were shown to have increased risk of advanced silent brain lesions and greater cardiac burden. PMID- 15127882 TI - Association between hepatocyte growth factor gene polymorphism and essential hypertension. AB - Hepatocyte growth factor (HGF) is a growth factor which contributes to protection and/or repair of vascular endothelial cells. Serum HGF level is elevated in response to hypertensive organ damage, which suggests that blood pressure regulation may be affected by HGF gene polymorphisms via serum HGF. To examine the interaction between a HGF gene polymorphism and hypertension, we carried out a case-control study. The present study was conducted in outpatients of Osaka University Hospital. Subjects (n=654) who gave informed consent to the study protocol and genetic analysis were recruited. A C to A nucleotide substitution in intron 13 of the HGF gene was determined by the TaqMan polymerase chain reaction (PCR) method using an MGB (Minor Groove Binder) probe. The genotype distribution of the C/A polymorphism of the HGF gene in total subjects was as follows: CC, 83%; CA, 16%; and AA 1%. This distribution was not significantly different from the predicted by Hardy-Weinberg's equilibrium. The prevalence of hypertension was significantly higher in subjects with the CC genotype than in those with an A allele, and the positive association remained after adjustment for confounding factors, with the estimated odds ratio for hypertension (CC vs. CA+AA) being 1.71 (95% confidence interval: 1.02-2.93). A significant association with hypertension was observed in lean or female subjects but not in obese or male subjects. In conclusion, our data suggested that C/A polymorphism in intron 13 of the HGF gene is associated with susceptibility to essential hypertension in lean or female subjects. PMID- 15127883 TI - Erectile dysfunction in hypertensive rats results from impairment of the relaxation evoked by neurogenic carbon monoxide and nitric oxide. AB - Erectile dysfunction (ED) with aging and diabetes mellitus is caused by impairment of the relaxation evoked by nitric oxide (NO) of penile cavernous smooth muscles and arterioles. However, the mechanism of ED in hypertension is unknown. Carbon monoxide (CO), which is produced by heme oxygenase (HO)-2 in the neuronal system is a neurotransmitter and a vasodilator. We examined the neurogenic role of CO in penile erection and the neurogenic mechanisms of ED in hypertension, using spontaneously hypertensive rats (SHR) or Wistar-Kyoto rats (WKY). The isometric tension of corpus cavernosum tissues from both strains was recorded after guanethidine and atropine treatment. Relaxation in response to electrical field stimulation (EFS) in WKY was suppressed dose-dependently by HO inhibitors both in the absence and presence of an NO synthase (NOS) inhibitor. Reverse transcription-polymerase chain reaction (RT-PCR) showed that the HO-2 gene was expressed in the corpus cavernosum. CO-saturated solution induced a concentration-dependent relaxation in WKY. The neurogenic relaxation to EFS in SHR was impaired as compared with that in WKY after the age of 5 weeks, when blood pressure began to be elevated, due to the attenuated relaxation in response to neurogenic NO and CO. In the corpus cavernosum of SHR, expression of the HO-2 and nNOS genes was similar, and NOx levels after EFS were similar to those of WKY. cGMP levels after EFS and the relaxation evoked by the NO donor was lower in SHR than WKY. Thiobarbituric acid-reacting substance (TBARS) levels were increased, and superoxide dismutase (SOD) activity was suppressed in SHR, as compared with those in WKY, suggesting that the increasing oxidative stress partially causes the impairment of NO-dependent relaxation. These findings suggest that CO regulates the relaxation evoked by EFS in the rat corpus cavernosum, and that ED in hypertension in rats results from an impairment of the relaxation induced by neurogenic CO and NO. PMID- 15127885 TI - Amlodipine and carvedilol prevent cytotoxicity in cortical neurons isolated from stroke-prone spontaneously hypertensive rats. AB - We previously reported that vitamin E prevents apoptosis in neurons during cerebral ischemia and reperfusion in stroke-prone spontaneously hypertensive rats (SHRSP). In this paper, we analyzed the effects of antihypertensives as well as vitamin E, which were added to neuron cultures after reoxygenation (20% O2) following hypoxia (1% O2). When added after hypoxia before reoxygenation, vitamin E conferred significant protection to neuronal cells. It was also shown that vitamin E conferred complete protection from neural cell death when added hypoxia and again before reoxygenation. At higher concentrations of vitamin E, strong neuroprotection was observed. Moreover, we verified that pretreatment with either amlodipine, carvedilol or dipyridamole consistently prevented cell death during hypoxia and reoxygenation (H/R). On the other hand, nilvadipine, a dihydropyridine-type calcium entry blocker, had no apparent effect on neuroprotection during H/R. The order of neuroprotective potency was vitamin E > dipyridamole > carvedilol > or = amlodipine > nilvadipine. In parallel experiments, we examined whether these antihypertensive agents were more effective when combined with vitamin E and dipyridamole. The results suggested that in our in vitro model system, antioxidants were the most important agents for the reduction of oxygen-free radical damage in cortical neurons. These findings suggest that amlodipine and carvedilol, with their antioxidant properties and antihypertensive activity, would be useful to inhibit neuronal cell death in the treatment of cerebrovascular stroke and neurodegenerative diseases in hypertensive patients. PMID- 15127884 TI - RhoA activation in vascular smooth muscle cells from stroke-prone spontaneously hypertensive rats. AB - RhoA is commonly activated in the aorta in various hypertensive models, indicating that RhoA seems to be a molecular switch in hypertension. The molecular mechanisms for RhoA activation in stroke-prone spontaneously hypertensive rats (SHRSP) were here investigated using cultured aortic smooth muscle cells (VSMC). The level of the active form of RhoA was higher in VSMC from SHRSP than in those from Wistar-Kyoto rats (WKY). The phosphorylation level of myosin phosphatase target subunit 1 (MYPT1) at the inhibitory site was also significantly higher in SHRSP, and the phosphorylation levels in both VSMCs were strongly inhibited to a similar extent by treatment with Y-27632, a Rho-kinase inhibitor. The expression levels of RhoA/Rho-kinase related molecules, namely RhoA, Rho-kinase, MYPT1, CPI-17 (inhibitory phosphoprotein for myosin phosphatase) and myosin light chain kinase, were not different between SHRSP and WKY. Valsartan, an angiotensin II (Ang II)- type 1 receptor antagonist, selectively and significantly reduced the RhoA activation in VSMC from SHRSP. The expression levels of the Rho GDP-dissociation inhibitor (RhoGDI) and leukemia associated Rho-specific guanine nucleotide exchange factor (RhoGEF) did not differ between SHRSP and WKY. In cyclic nucleotide signaling, cyclic GMP (cGMP) dependent protein kinase Ialpha (cGKIalpha) was significantly downregulated in SHRSP cells, although there were no changes in the expression levels of guanylate cyclase beta and cyclic AMP (cAMP)-dependent protein kinase or the intracellular contents of cGMP and cAMP between the two rat models. These results suggest that the possible mechanisms underlying RhoA activation in VSMC from SHRSP are autocrine/paracrine regulation by Ang II and/or cGKIalpha downregulation. PMID- 15127886 TI - Cyclin-dependent kinase inhibitor, p21Waf1, regulates vascular smooth muscle cell hypertrophy. AB - In the process of vascular diseases, smooth muscle cells (SMC) undergo not only hyperplasia but also hypertrophy, resulting in vascular remodeling. A cyclin dependent kinase inhibitor (CDKI), p21Waf1, has been shown to play an important role in SMC hyperplasia. Here we investigated a potential role of p21Waf1 in SMC hypertrophy. An exposure of cultured rat SMC to serum drove the cell cycle progression with up-regulation of various cell cycle markers and increased activities of cyclin-dependent kinases, but did not cause SMC hypertrophy. In contrast, incubation of SMC for 48 h with angiotensin II (AII, 100 nmol/l) resulted in a significant increase in the cell size measured by flowcytometric forward-angle light scatter assay, in association with an increase in the ratio of [3H]leucine/[3H]thymidine uptake, indicating SMC hypertrophy. At 48 h, p21Waf1 expression was up-regulated in SMC exposed to AII but not in those exposed to serum. These results suggest that p21Waf1 may be involved in hypertrophy. To further investigate this issue, two manipulations of the p21Waf1 gene were performed. Adenovirus-mediated over-expression of p21Waf1 not only reduced S phasic cells but also caused hypertrophy, despite the exposure to serum. Antisense oligodeoxynucleotide for p21Waf1 inhibited the hypertrophy of SMC exposed to AII. Our data suggest that p21Waf1 may play a role in SMC hypertrophy as well. PMID- 15127887 TI - Olmesartan medoxomil, an angiotensin II receptor blocker ameliorates insulin resistance and decreases triglyceride production in fructose-fed rats. AB - Although angiotensin II receptor blockers (ARBs) have been recommended as a first line of anti-hypertensive agents in patients with diabetes, it remains unclear whether ARBs have a favorable effect on insulin action and triglyceride (TG) metabolism, both of which are impaired in type 2 diabetes. In this study we addressed this issue by investigating how a newly developed ARB, olmesartan medoxomil, influenced insulin sensitivity and TG metabolism in fructose-fed rats, a representative animal model of insulin resistance. Olmesartan was administrated as a 0.01% drinking solution ad libitum to rats either fed normal chow or fructose-enriched chow (60%) for 21 days. Olmesartan treatment markedly decreased both systolic and diastolic blood pressure in both chow-fed and fructose-fed animals. The area under the curve of insulin (AUCI) was substantially greater in fructose-fed rats in the intravenous glucose tolerance test, and olmesartan treatment significantly reduced the AUCI. Olmesartan significantly improved the insulin sensitivity index in fructose-fed rats assessed by Bergman's minimal model without affecting insulin-independent glucose disposal. Olmesartan significantly decreased plasma TG and non-esterified fatty acid levels in fructose-fed rats without affecting lipoprotein lipase mass. The TG secretion rate determined by the triton WR1339 technique was two-fold higher in fructose fed rats, but olmesartan restored the TG secretion to a normal rate. Olmesartan did not affect plasma parameters, insulin sensitivity or TG metabolism in chow fed rats. Olmesartan ameliorates insulin resistance and overproduction of TG in fructose-fed rats, and these effects appear to be independent of its hypotensive action. PMID- 15127888 TI - Statistical analysis of oligonucleotide microarray data. AB - Microchip arrays have become one of the most rapidly growing techniques for monitoring gene expression at the genomic level and thereby gaining valuable insight about various important biological mechanisms. Examples of such mechanisms are: identifying disease-causing genes, genes involved in the regulation of some aspect of the cell cycle, etc. In this article, we discuss the problem of estimating gene expression based on a proper statistical model. More precisely, we show how the model introduced by Li and Wong can be used in its full bivariate generality to provide a new measure of gene expression from high density oligonucleotide arrays. We also present a second gene expression index based on a new way of reducing the model into a simpler univariate model. In both cases, the gene expression indices are shown to be unbiased and to have lower variance than the established ones. Moreover, we present a bootstrap method aiming at providing non-parametric confidence intervals for the expression index. PMID- 15127889 TI - Simulation-based estimation of stochastic process parameters in tumor growth. AB - Models are generally developed at the micro level. Data are generally gathered at the macro level. Obtaining the macromodel which is the natural consequence of the underlying micro model is generally not feasible. SIMEST gives a means whereby the micromodel is used to generate, for a given assumed set of parameters, simulated sets of macro data. These data are compared with the actual clinical macro data. The parameters are then adjusted to obtain concordance with the clinical data. In this manner, simulation gives us a means of parameter estimation without the necessity of generating the macro model. PMID- 15127890 TI - Cell cycle progression. AB - In this paper we consider cell cycle models for which the transition operator for the evolution of birth mass density is a simple, linear dynamical system with a stochastic perturbation. The convolution model for a birth mass distribution is presented. Density functions of birth mass and tail probabilities in n-th generation are calculated by a saddle-point approximation method. With these probabilities, representing the probability of exceeding an acceptable mass value, we have more control over pathological growth. A computer simulation is presented for cell proliferation in the age-dependent cell cycle model. The simulation takes into account the fact that the age-dependent model with a linear growth is a simple linear dynamical system with an additive stochastic perturbation. The simulated data as well as the experimental data (generation times for mouse L) are fitted by the proposed convolution model. PMID- 15127891 TI - Bifurcations in a white-blood-cell production model. AB - We study the dynamics of a model of white-blood-cell (WBC) production. The model consists of two compartmental differential equations with two discrete delays. We show that from normal to pathological parameter values, the system undergoes supercritical Hopf bifurcations and saddle-node bifurcations of limit cycles. We characterize the steady states of the system and perform a bifurcation analysis. Our results indicate that an increase in apoptosis rate of either hematopoietic stem cells or WBC precursors induces a Hopf bifurcation and an oscillatory regime takes place. These oscillations are seen in some hematological diseases. PMID- 15127892 TI - Modeling operon dynamics: the tryptophan and lactose operons as paradigms. AB - Understanding the regulation of gene control networks and their ensuing dynamics will be a critical component in the understanding of the mountain of genomic data being currently collected. This paper reviews recent mathematical modeling work on the tryptophan and lactose operons which are, respectively, the classical paradigms for repressible and inducible operons. PMID- 15127893 TI - Using mathematical models to help understand biological pattern formation. AB - One of the characteristics of biological systems is their ability to produce and sustain spatial and spatio-temporal pattern. Elucidating the underlying mechanisms responsible for this phenomenon has been the goal of much experimental and theoretical research. This paper illustrates this area of research by presenting some of the mathematical models that have been proposed to account for pattern formation in biology and considering their implications. PMID- 15127894 TI - Contribution to the study of periodic chronic myelogenous leukemia. AB - The period (in the order of 40 to 80 days) in periodic chronic myelogenous leukemia (PCML) oscillations is quite long compared with the duration of the cell cycle of the hematopoietic stem cells from which the oscillations are presumed to originate (in the order of one or two days). Our objective is to understand the origin of these long-period oscillations using a G0 model for stem cell dynamics. We determine the local stability conditions and show under what conditions the Hopf bifurcation may occur. We interpret the role of each parameter in the loss of stability, and then examine a simpler model to try to deduce possible changes at the stem-cell level that might be responsible for the characteristics PCML. PMID- 15127895 TI - A mathematical derivation of size spectra in fish populations. AB - Taking into account a predator/prey size ratio in a size-structured population model leads to a partial derivative equation of which we study the properties. By expliciting the structure of the attractor of this equation, it is shown that a simple mechanism, size-based opportunistic predation, can explain the stability in the shape of size spectra observed in various marine ecosystems. PMID- 15127896 TI - Modelling individual plant growth at a variable mean density or at a specific spatial setting. AB - Neighbouring plants generally compete for the limiting resources in order to grow and reproduce. Some resources, e.g., sun light, may be monopolised by the larger plants and this may lead to asymmetric competition where a plant, which is twice as large, grows more than twice as fast. A previously published individual-based Richards growth model that describes the asymmetric growth of individual plants is here generalised with respect to a variable mean plant density and an explicit spatial setting. PMID- 15127897 TI - Individual-based spatially-explicit model of an herbivore and its resource: the effect of habitat reduction and fragmentation. AB - We present an individual-based, spatially-explicit model of the dynamics of a small mammal and its resource. The life histories of each individual animal are modeled separately. The individuals can have the status of residents or wanderers and belong to behaviorally differing groups of juveniles or adults and males or females. Their territory defending and monogamous behavior is taken into consideration. The resource, green vegetation, grows depending on seasonal climatic characteristics and is diminished due to the herbivore's grazing. Other specifics such as a varying personal energetic level due to feeding and starvation of the individuals, mating preferences, avoidance of competitors, dispersal of juveniles, as a result of site overgrazing, etc., are included in the model. We determined model parameters from real data for the species Microtus ochrogaster (prairie vole). The simulations are done for a case of an enclosed habitat without predators or other species competitors. The goal of the study is to find the relation between size of habitat and population persistence. The experiments with the model show the populations go extinct due to severe overgrazing, but that the length of population persistence depends on the area of the habitat as well as on the presence of fragmentation. Additionally, the total population size of the vole population obtained during the simulations exhibits yearly fluctuations as well as multi-yearly peaks of fluctuations. This dynamics is similar to the one observed in prairie vole field studies. PMID- 15127898 TI - Time delay as a key factor of model plankton dynamics. AB - Studies of the mechanisms underlying complex dynamics of ecological systems at various spatial and time scales bring increasing awareness that complexity is an intrinsic feature of ecological functioning. This paper is to investigate the role of such an ecologically significant parameter as the time delay due to maturation processes in the complex plankton dynamics. We show that the time lag T1, associated with the zooplankton maturation period can lead to essential changes in the plankton dynamics. Particularly, we show that the coexistence of limit cycle and chaotic attractor we have recently found to be typical of the system at T1 = 0 [A.B. Medvinsky, I.A. Tikhonova, R.R. Aliev, B.-L. Li, Z.-S. Lin, H. Malchow, Patchy environment as a factor of complex plankton dynamics, Phys. Rev. E 64 (2001) 021915] is replaced by pure chaotic plankton dynamics as T1 becomes more than a critical value. The results obtained imply that chaos is a rather common phenomenon in the plankton functioning. PMID- 15127899 TI - Effect of variable surrounding on species creation. AB - We constructed a model of speciation from evolution in an ecosystem consisting of a limited amount of energy recources. The species possesses genetic information, which is inherited according to the rules of the Penna model of genetic evolution. The increase in the number of the individuals of each species depends on the quality of their genotypes and the available energy resources. The decrease in number of the individuals results from genetic death or maximum-age reaching by the individual. The amount of energy resources is represented by a solution of the differential logistic equation, where the growth rate of the amount of the energy resources has been modified to include the number of individuals from all species in the ecosystem under consideration. The fluctuating surrounding is modelled with the help of the function V(x, t) = 1/4 x4 + 1/2 b(t)x2, where x represents phenotype and the coefficient b(t) shows the cos(omega t) time dependence. The closer the value x of an individual to the minimum of V(x, t), the better adapted its genotype to the surrounding. We observed that the life span of the organisms strongly depends on the value of the frequency omega. It becomes shorter the more frequent the changes of the surrounding. However, there is a tendency for the species that have a higher value of the reproduction age aR to win the competition with the other species. Another observation is that small evolutionary changes of the inherited genetic information lead to spontaneous bursts of the evolutionary activity when many new species may appear in a short period. PMID- 15127900 TI - A numerical simulation for the dynamics of the sexual phase of monogonont rotifera. AB - A numerical simulation for the dynamics of a model that describes the sexual phase of Monogonont Rotifera reproduction is presented. The simulation is carried out by means of a numerical method based on the integration along the characteristic curves. The numerical experiments cover two basic situations: the existence of an asymptotic stable equilibrium state and the existence of an stable periodic solution. Our results are in agreement with the theoretical analysis made by Calsina and Ripoll (J. Math. Biol. 45 (2002) 22). PMID- 15127901 TI - War and anxiety disorders. PMID- 15127902 TI - Susceptible subgroups: the challenge of studying interactions. PMID- 15127903 TI - Male circumcision and AIDS in Africa: Primum non nocere versus the collective good. PMID- 15127904 TI - Gulf War veterans with anxiety: prevalence, comorbidity, and risk factors. AB - BACKGROUND: Veterans of the first Gulf War have higher rates of medical and psychiatric symptoms than nondeployed military personnel. METHODS: To assess the prevalence of and risk factors for current anxiety disorders in Gulf War veterans, we administered a structured telephone interview to a population-based sample of 4886 military personnel from Iowa at enlistment. Participants were randomly drawn from Gulf War regular military, Gulf War National Guard/ Reserve, non-Gulf War regular military, and non-Gulf War National Guard/Reserve. Medical and psychiatric conditions were assessed through standardized interviews and questionnaires in 3695 subjects (76% participation). Risk factors were assessed using multivariate logistic regression models. RESULTS: Veterans of the first Gulf War reported a markedly higher prevalence of current anxiety disorders than nondeployed military personnel (5.9% vs. 2.8%; odds ratio = 2.1; 95% confidence interval = 1.3-3.1), and their anxiety disorders are associated with co-occurring psychiatric disorders. Posttraumatic stress disorder, panic disorder, and generalized anxiety disorder were each present at rates nearly twice expected. In our multivariate model, predeployment psychiatric treatment and predeployment diagnoses (posttraumatic stress disorder, depression, or anxiety) were independently associated with current anxiety disorder. Participation in Gulf War combat was independently associated with current posttraumatic stress disorder, panic disorder, and generalized anxiety disorder. CONCLUSIONS: Current anxiety disorders are relatively frequent in a military population and are more common among Gulf War veterans than nondeployed military personnel. Predeployment psychiatric difficulties are robustly associated with the development of anxiety. Healthcare providers and policymakers need to consider panic disorder and generalized anxiety disorder, in addition to posttraumatic stress disorder, to ensure their proper assessment, treatment, and prevention in veteran populations. PMID- 15127905 TI - Who is sensitive to the effects of particulate air pollution on mortality? A case crossover analysis of effect modifiers. AB - BACKGROUND: Populations susceptible to the effects of particulate matter have begun to be characterized, but the independent contributions of specific factors have not been explored. METHODS: We used a case-crossover study to examine PM10 associated mortality risk during 1988-1991 among 65,180 elderly residents of Cook County, Illinois, who had a history of hospitalization for heart or lung disease. We assessed how the effect was independently modified by specific diagnoses and personal characteristics. RESULTS: We found a 1.14% (95% confidence interval [CI] = 0.44% to 1.85%) increased risk of death per 10 microg/m3 increase in ambient PM10 concentration. Persons with heart or lung disease-but no specific diagnosis of myocardial infarction, diabetes, congestive heart failure, chronic obstructive pulmonary disorder, or conduction disorders-were at 0.74% (-0.29% to 1.79%) increased risk. Persons with a history of myocardial infarction had a 2.7-fold higher risk (CI = -2.1 to 7.4). Those with diabetes carried a 2.0-fold higher risk (CI = -1.5 to 5.5). Risk appeared to decrease with age among elderly men and increase with age among elderly women, but the estimated 3-way interaction was not precise enough to exclude the null. We found no indication that susceptibility varied by group-level socioeconomic measures. CONCLUSION: Among a frail population, individuals diagnosed with myocardial infarction or diabetes were at greatest risk of death associated with high concentrations of PM10. These results suggest that their susceptibility may derive from prior vascular damage to the heart. PMID- 15127906 TI - Maternal smoking and the risk of orofacial clefts: Susceptibility with NAT1 and NAT2 polymorphisms. AB - BACKGROUND: Orofacial clefts are etiologically heterogeneous malformations. One probable cause is maternal smoking during pregnancy. The effect of maternal smoking may be modified by genes involved in biotransformation of toxic compounds derived from tobacco. We investigated whether polymorphic variants of fetal acetyl-N-transferases 1 (NAT1) and 2 (NAT2) interact with maternal cigarette smoking during early pregnancy to increase the risk of delivering an infant with an orofacial cleft. METHODS: In a California population-based case-control study, we genotyped 421 infants born with an isolated cleft and 299 nonmal-formed controls for 2 NAT1 and 3 NAT2 single nucleotide polymorphisms RESULTS: Although smoking was independently associated with increased risks for both isolated cleft lip +/- cleft palate and isolated cleft palate, no independent associations were found for NAT1 1088 or 1095 genotypes or for NAT2 acetylator status. However, the infant NAT1 1088 and 1095 polymorphisms were strongly associated with the risk of clefts among smoking mothers; infants with NAT1 1088 genotype AA versus TT (odds ratio [OR] = 3.9; 95% confidence interval = 1.1-17.2) and with NAT1 1095 genotype AA versus CC (OR = 4.2; 1.2-18.0). Infant NAT2 acetylator status did not appreciably affect susceptibility of the fetus to the teratogenic effects of maternal smoking. CONCLUSIONS: Our results suggest that maternal smoking during pregnancy may increase risk for orofacial clefts particularly among smokers whose fetuses have polymorphic variants of NAT1, an enzyme involved in phase II detoxification of tobacco smoke constituents. PMID- 15127907 TI - Risk of HIV-1 in rural Kenya: a comparison of circumcised and uncircumcised men. AB - BACKGROUND: Most studies that have found an association between uncircumcised status and infection with human immunodeficiency virus type 1 (HIV-1) have compared participants from various demographic backgrounds, among which the prevalence of other risk factors might have varied. We report findings from a study conducted among men within a single ethnic community in which circumcision was dictated by the religious denomination to which the men belonged. METHODS: Of the 1217 eligible men, we included in the analysis 845 who gave blood samples for HIV-1 testing and who were confirmed as either fully circumcised (n = 398) or uncircumcised (n = 447). The seroprevalence of HIV-1 was compared between the 2 groups. RESULTS: All correlates of HIV-1 prevalence that we measured were distributed similarly between circumcised and uncircumcised men. The seroprevalence of HIV-1 was 30% among the uncircumcised men and 20% among the circumcised men. Among uncircumcised men, HIV-1 seroprevalence was similar between men from circumcising denominations (31%; n = 111) and noncircumcising denominations (30%; n = 336). The crude prevalence ratio for HIV infection associated with not being circumcised was 1.5 (95% confidence interval = 1.2 2.0); and adjustment for other measured risk factors for HIV-1 infection had little impact on this result. CONCLUSION: Our study provides evidence that circumcision is associated with a reduced risk of HIV-1 infection. PMID- 15127908 TI - Estimating HIV incidence and detection rates from surveillance data. AB - BACKGROUND: Markov models that incorporate HIV test information can increase precision in estimates of new infections and permit the estimation of detection rates. The purpose of this study was to assess the functioning of a Markov model for estimating new HIV infections and HIV detection rates in Louisiana using surveillance data. METHODS: We expanded a discrete-time Markov model by accounting for the change in AIDS case definition made by the Centers for Disease Control and Prevention in 1993. The model was applied to quarterly HIV/AIDS surveillance data reported in Louisiana from 1981 to 1996 for various exposure and demographic subgroups. When modeling subgroups defined by exposure categories, we adjusted for the high proportion of missing exposure information among recent cases. We ascertained sensitivity to changes in various model assumptions. RESULTS: The model was able to produce results consistent with other sources of information in the state. Estimates of new infections indicated a transition of the HIV epidemic in Louisiana from (1) predominantly white men and men who have sex with men to (2) women, blacks, and high-risk heterosexuals. The model estimated that 61% of all HIV/AIDS cases were detected and reported by 1996, yet half of all HIV/non-AIDS cases were yet to be detected. Sensitivity analyses demonstrated that the model was robust to several uncertainties. CONCLUSIONS: In general, the methodology provided a useful and flexible alternative for estimating infection and detection trends using data from a U.S. surveillance program. Its use for estimating current infection will need further exploration to address assumptions related to newer treatments. PMID- 15127909 TI - Does appendectomy reduce the risk of ulcerative colitis? AB - BACKGROUND: Persons who have had an appendectomy are less prone to develop ulcerative colitis than those who have not. The underlying mechanism and the clinical use of this finding are unknown. METHODS: Two competing hypotheses were examined: (1) the inverse association between appendectomy and ulcerative colitis is conferred by a mutual determinant such as genes, and (2) the appendix itself has biologic effects that promote the development of ulcerative colitis. We performed a population-based cohort study of all 234,559 persons who had an appendectomy performed in Denmark 1977 through 1999. We used a bidirectional cohort design, calculating the standardized incidence rate of ulcerative colitis both before and after the appendectomy. If the hypothesis of a constant, confounding factor were true, incidence rates of ulcerative colitis would be equal before and after appendectomy. If the incidence of ulcerative colitis were lower after appendectomy than before, it would support the second hypothesis and a true protective effect of the appendectomy. We controlled confounding by age and sex using direct standardization. RESULTS: Of 234,559 persons who had an appendectomy, 559 developed ulcerative colitis during a mean follow up of 17.5 years. The standardized incidence rate of ulcerative colitis was lower in the postappendectomy period than in the preappendectomy period (incidence rate ratio = 0.74; 95% confidence interval = 0.62-0.88). CONCLUSIONS: The inverse association between appendectomy and ulcerative colitis is not explained by time stable factors such as genes. Appendectomy might have a genuine protective effect toward development of ulcerative colitis. PMID- 15127910 TI - Trihalomethanes in public water supplies and risk of stillbirth. AB - BACKGROUND: The chlorine used to disinfect public drinking water supplies reacts with naturally occurring organic matter to form a number of chemical byproducts. Recent studies have implicated exposure to chlorination byproducts in drinking water, trihalomethanes (THMs), in particular, with intrauterine death. METHODS: We conducted a population-based case-control study in Nova Scotia and Eastern Ontario, Canada, to examine the effect of exposure to THMs on stillbirth risk. Cases were women who had a stillborn infant, and controls were a random sample of women with live births. Subjects were interviewed, and women with a public water source provided a residential water sample. Risks were examined according to residential THM level in tap water and to a total exposure metric incorporating tap water ingestion, showering, and bathing. RESULTS: We enrolled 112 stillbirth cases and 398 live birth controls. Women with a residential total THM level of 80 or more microg/L had twice the risk of a stillbirth compared with women with no exposure to THMs (adjusted odds ratio [OR] = 2.2; 95% confidence interval [CI] = 1.1-4.4). The highest quintile of total THM exposure using the total exposure metric was associated with an adjusted odds ratio of 2.4 (95% CI = 1.2-4.6) compared with women not exposed to THMs. Similar results were seen for specific THM compounds. A monotonic dose-response relationship was not seen. CONCLUSIONS: Our results provide evidence for an increased risk of stillbirth associated with exposure to chlorination byproducts through ingestion and showering and bathing, although there was not a clear dose-response relationship. PMID- 15127911 TI - Farm children's exposure to herbicides: comparison of biomonitoring and questionnaire data. AB - BACKGROUND: Pesticide exposure has been associated with various childhood cancers. However, most studies rely on questionnaires, with few using biologic measures of dose. This study was designed to measure herbicide exposure directly in children of farm applicators, and to compare these results with exposure imputed from questionnaire information. METHODS: Two consecutive 24-hour urine samples were collected from 92 children of Ontario farm applicators who used the herbicides 2,4-D (2,4-dichlorophenoxyacetic acid) or MCPA (4-chloro-2 methylphenoxyacetic acid) for the first time during 1996. The farm applicator completed questionnaires describing his pesticide-handling practices as well as the child's location during the various stages of handling these pesticides. RESULTS: Approximately 30% of the children on farms using these herbicides had detectable concentrations in their urine, with maximum values of 100 microg/L for 2,4-D and 45 microg/L for MCPA. Children with higher levels were more likely to be boys and to have parents who also had higher mean urinary concentrations. The sensitivity and specificity of a simple indicator of use were 47% and 72%, respectively, for 2,4-D, and 91% and 30%, respectively, for MCPA, using the biomonitoring data as the gold standard. CONCLUSIONS: Information on living on a farm, or on living on a farm where a specific pesticide is used, is not enough to classify children's exposures. Given this potential for misclassification, we urge incorporation of biomonitoring studies in subsets of children at least to estimate the extent of misclassification. PMID- 15127912 TI - Fetal trisomy 21 and maternal preeclampsia. AB - BACKGROUND: Placental trophoblast shedding into maternal circulation has been hypothesized as a potential cause of preeclampsia. Because pregnancies with a trisomy 21 fetus also have high levels of fetal cells and cell-free fetal DNA in maternal circulation, we examined whether trisomy 21 pregnancies have a higher risk of preeclampsia than euploid pregnancies. METHODS: We used 2 population based databases. We identified 7763 pregnancies with a singleton trisomy 21 affected fetus and 15,293 matched euploid gestations from the U.S. Natality files for the period 1995-1999. The second database consisted of 665 pregnancies with fetal trisomy 21 and 987 euploid controls in a population-based Down syndrome study in California. In the latter study, women were interviewed by telephone regarding characteristics and pregnancy complications. Gestational hypertension and preeclampsia are the outcomes of this study. RESULTS: The U.S. Natality files showed that in nulliparous women fetal trisomy 21 was associated with a reduced risk of pregnancy-induced hypertension (adjusted relative risk [aRR] = 0.67; 95% confidence interval [CI] = 0.53 to 0.85). Findings from the California study confirmed this association in nulliparous women, and further revealed that the decrease in overall risk of pregnancy-induced hypertension was mainly the result of a large reduction in the risk of preeclampsia (aRR = 0.19; CI = 0.04 to 0.88) rather than in gestational hypertension by itself (0.83; 0.37 to 1.84). Neither dataset showed these effects among multiparous pregnancies. CONCLUSION: Fetal trisomy 21 is associated with a reduced, rather than increased, risk of preeclampsia, specifically in nulliparous women. PMID- 15127913 TI - Quantifying delay in access to new medical treatment: an application of risk advancement period methodology. AB - BACKGROUND: We present a novel application of the concept of risk or rate advancement to compute the extent of delay in adoption of an effective new drug in 2 German health insurance systems. METHODS: We identified individuals with migraines, age 18 to 65 years, in 371 primary care practices in Germany in 1994 (MediPlus, IMS Health database). These included 8173 persons covered under the statutory health insurance system and 503 persons covered by private health insurance. We derived risk and population risk advancement periods for sumatriptan compared with nonserotoninergic acute migraine therapy using multiplicative risk regression and generalized estimating equations, adjusted for patient, physician, and practice cofactors. RESULTS: For patients at the mean age of the cohort, 43 years of age, sumatriptan was prescribed 1.2 (95% confidence interval [CI] = 0.3-2.0) years later among those in the statutory health insurance system compared with those who had private insurance. The lag increased by 0.6 (-0.1 to 1.3) years for every 10 years of patient age. In the age-mix of our sample, access to the health benefits of sumatriptan therapy lagged nearly 1.5 years behind in the statutory health insurance system and for Germany as a whole. CONCLUSIONS: Migraine patients' access to sumatriptan therapy lagged substantially in the statutory health insurance system and in the country as a whole. Risk advancement periods provide a useful methodology for communicating major healthcare issues in a meaningful way to society and policymakers. PMID- 15127914 TI - Incidence of thrombotic thrombocytopenic purpura/hemolytic uremic syndrome. AB - BACKGROUND: Thrombotic thrombocytopenic purpura and hemolytic uremic syndrome are rare disorders characterized by platelet aggregation, microthrombi, and resulting tissue damage. We studied the incidence and possible risk factors for these diseases in 3 large populations in the United States, United Kingdom, and Canada. METHODS: Data were derived from a large health insurer in the United States, general practices in the United Kingdom, and the Province of Saskatchewan. We identified potential cases of thrombotic thrombocytopenia purpura and hemolytic uremic syndrome in computerized data and verified them by medical record review. We estimated incidence rates for thrombotic thrombocytopenia purpura and hemolytic uremic syndrome together and separately, and we conducted a case control study to evaluate potential risk factors. RESULTS: The age-sex standardized incidence of thrombotic thrombocytopenia purpura and hemolytic uremic syndrome was higher than previously reported (6.5, 2.2, and 3.2 per million per year in the United States, United Kingdom, and Saskatchewan, respectively), but there was no secular trend. The incidence of thrombotic thrombocytopenia purpura and hemolytic uremic syndrome was higher in women than men. Most cases of hemolytic uremic syndrome occurred before 20 years of age. We confirmed several known risk factors for thrombotic thrombocytopenia purpura and hemolytic uremic syndrome (cancer, bone marrow transplantation, pregnancy). CONCLUSION: The incidence of thrombotic thrombocytopenia purpura and hemolytic uremic syndrome is higher than previously reported but does not appear to be rising. Apparent international differences in incidence could be the result of imprecision in identifying thrombotic thrombocytopenia purpura and hemolytic uremic syndrome in large research databases. PMID- 15127915 TI - On food frequency questionnaires: the contribution of open-ended questions and questions on ethnic foods. AB - BACKGROUND: Epidemiologic studies of diet must balance the goal of comprehensiveness with that of minimizing subject and study burden. Information on the impact of certain additional questionnaire elements may aid in such decisions. METHODS: We administered a food-frequency questionnaire in a large multiethnic cohort of women (Study of Women's Health Across the Nation) in 1996 1997. Ancillary questionnaires asked about "any other foods you eat at least once a week" and about consumption of 8-16 ethnic foods included for respondents of Hispanic, Chinese, or Japanese ethnicity. We assessed the impact of these 2 ancillary questionnaires by examining the mean nutrient amount they contributed and their effects on rank-order correlations and categorizations of nutritional estimates. We examined the impact of ethnic foods both within the relevant ethnic group and among non-Hispanic whites. RESULTS: Rank-order correlations (rs) between nutrient estimates with and without the open-ended question were 0.99 for almost all nutrients and all ethnic groups. Nutrient amounts added by the open ended question rarely exceeded 2% of the total. Weighted kappa statistics for quintile classifications were greater than 0.9. Similarly, asking white respondents about ethnic foods added little to nutrient estimates, and all correlations between estimates with and without the ethnic foods were at least 0.95. The effect of ethnic foods on nutrient means within the relevant ethnic group was more substantial, although most correlations between estimates with and without the added foods were still above 0.9. CONCLUSIONS: Open-ended food questions added little to nutrient estimates or rankings in any of the ethnic groups. Specific questions about ethnic foods were useful only within ethnic groups, for which they increased mean nutrient estimates and had modest impact on ranking. PMID- 15127916 TI - Changes in alcohol intake and mortality: a longitudinal population-based study. AB - BACKGROUND: Using alcohol intake at one point in time, numerous studies have shown a J- or U-shaped relation with all-cause mortality. Mortality is lowest among the light to moderate drinkers, with the risk of dying from coronary heart disease higher among nondrinkers and the risk of dying from cancer higher among heavy drinkers. We studied whether changes in individual alcohol intake result in corresponding changes in mortality. METHODS: In a longitudinal study of 6644 men and 8010 women, age 25 to 98 years, who had attended at least 2 health surveys with a 5-year interval between them, we addressed the risk of death after combinations of changes in alcohol intake. RESULTS: Mortality after changes in alcohol intake was consistent with the mortality observed among those who reported stable drinking. Stable drinkers showed a U-shaped all-cause mortality, with relative risks of 1.29 (95% confidence interval [CI] = 1.13-1.48) for nondrinkers (< 1 drink per week) and 1.32 (1.15-1.53) for heavy drinkers (> 13 drinks per week) compared with light drinkers (1 to 6 drinks per week). For coronary heart disease mortality, stable nondrinkers had a relative risk of 1.32 (0.97-1.79) compared with stable light drinkers and those who had reduced their drinking from light to none increased their risk (1.40; 1.00-1.95), and those who had increased from nondrinking to light drinking reduced their relative risk ratio (0.71; 0.44-1.14). Cancer mortality was increased in all groups of heavy drinkers. CONCLUSION: Persons with stable patterns of light and moderate alcohol intake had the lowest all-cause mortality. Individual changes in alcohol intake were followed by corresponding changes in mortality. PMID- 15127917 TI - Maternal caffeine consumption and spontaneous abortion: a review of the epidemiologic evidence. AB - Since the 1980s, numerous studies have investigated whether caffeine intake during pregnancy affects the risk of spontaneous abortion. A clear consensus, however, has not been reached. Because of the public health importance of this question, we reviewed the results of 15 epidemiologic studies on this topic, with particular attention to the specific methodologic problems that would generate biased findings. These include selection and recall bias, confounding, several issues pertaining to exposure measurement, and the failure to account for fetal karyotype, caffeine metabolism, the timing of fetal demise, and the possibility that an effect of caffeine may be gestational age-specific. All of the studies reviewed suffer from important methodologic limitations that hinder both the interpretation of each study individually and the comparison of results across studies. Despite the fact that most epidemiologic studies have observed a positive association between maternal caffeine intake and the risk of spontaneous abortion, we conclude that the evidence must be considered to be equivocal, given the biases likely present and the fact that most of the potential biases would tend to overestimate any association. Until studies can overcome these limitations, evidence for a causal link between caffeine and spontaneous abortion will remain inconclusive. PMID- 15127919 TI - A conversation with Margaret Becklake. PMID- 15127918 TI - Who is Hispanic? Implications for epidemiologic research in the United States. AB - The most recent U.S. Census reported that Hispanics are now the nation's largest minority group. At the same time, increasing attention has focused on the inherent heterogeneity of the U.S. Hispanic population. Such a rapidly growing but heterogeneous minority poses potential challenges to population-based research. To understand those challenges better, we first considered the history of the demographers' question: "Who is Hispanic?" We then considered the implications of differing Hispanic identity criteria for disease surveillance. Although relevant to political and socioeconomic considerations, the Hispanic ethnic category may not be specifically useful for understanding most disease processes. For epidemiologic studies, there is need for more transparent criteria to classify subpopulations. Those criteria must be regularly subjected to analysis and validation. PMID- 15127920 TI - "Looking old for your age": genetics and mortality. PMID- 15127921 TI - Body mass index and preeclampsia. PMID- 15127922 TI - Occupational exposures and neurodegenerative diseases. PMID- 15127923 TI - Arsenic and drinking water. PMID- 15127924 TI - Not the last bifocal trial! PMID- 15127925 TI - Development and validation of a multidimensional quality-of-life scale for myopia. AB - BACKGROUND: Five dimensions of health-related quality of life in myopia were hypothesized to affect satisfaction with visual correction modality. Items on these dimensions reflected the frequency of visual compromise and ocular symptoms; individual tolerance of these compromises and symptoms; cosmesis; psychological constructs (including situation-dependent characteristics such as adaptability, self-efficacy, and subjective well-being); and personality traits such as extraversion and introversion. METHODS: Psychologically oriented items and visually oriented items were developed in two stages involving 1,647 participants. Item development was based on a comprehensive literature review, interviews with experts, myopic subjects, and graduate students, and written feedback. Items were selected through factor analysis and the examination of their ability to discriminate between treatment conditions (spectacle wear, daily use of contact lenses, continuous use of contact lenses, or laser-assisted in situ keratomileusis for myopia). After development, selection, and validation of the psychological items and then the vision items, a final multidimensional scale combining both types of items was designed. The scale was administered to 124 subjects whose myopia was corrected by one of several treatment modalities to determine final construct validity. RESULTS: Using principal axis factoring and oblimin with Kaiser normalization rotation methods, five factors with strong item loadings evolved as hypothesized. The final multidimensional scale consisted of 13 items related to specific aspects of frequency of visual compromise and ocular symptoms with 13 corresponding items for level of tolerance for these problems; three items related to cosmesis; 10 items related to psychological characteristics; and six items related to personality traits. Good internal consistency in each factor (Cronbach's alpha range, 0.76 to 0.92) for the scale was evident. DISCUSSION: This report describes the development and validation of an easily administered, short, effective multidimensional health-related quality of-life questionnaire for use in selecting and measuring success of methods for correcting myopia. PMID- 15127926 TI - Use of the experience sampling method to measure nearwork. AB - PURPOSE: The purpose of this study was to use the Experience Sampling Method (ESM) to evaluate the daily visual activities of a group of optometry students and to determine whether the ESM could detect differences in visual activities when a difference was thought to exist. METHODS: Sixty-two optometry students, at four different levels of training, at The Ohio State University College of Optometry were tested using the ESM during a 14-day period. A subgroup of 18 subjects in their fourth year of study was asked to repeat the 14-day ESM on a second occasion. The two ESM periods were specifically chosen at times when the subjects were expected to have markedly different nearwork profiles. RESULTS: A comparison of members from the four optometry classes when the fourth-year students were taking final written examinations showed no significant differences in the proportion of nearwork among groups (p = 0.170); however, when the fourth year students were primarily performing eye examinations during clinical rotations, there was a significant difference in the proportion of nearwork among training levels (13.8% for fourth-year students compared with 24.9%, 23.7%, and 30.4% for the other optometry classes; p = 0.0001). The percentage of time spent doing nearwork and performing eye examinations was significantly different between two ESM periods (paired t-test, p = 0.0001 for each activity). CONCLUSIONS: The ESM is capable of detecting differences in the proportion of time spent doing nearwork and performing eye examinations among groups of optometry students in different levels of training with different daily visual demands and between two ESM periods when these activities were expected to differ. The ESM can be used as an alternative method for nearwork quantification in future studies. PMID- 15127927 TI - Prevalence of myopia in a group of Hong Kong microscopists. AB - PURPOSE: To study the prevalence and magnitude of myopia in a group of Hong Kong Chinese microscopists and compare it with that observed in microscopists working in the United Kingdom. METHODS: Forty-seven microscopists (36 women and 11 men) with a median age of 31 years and working in hospital laboratories throughout Hong Kong were recruited to the study. Information about past refractive corrections, microscopy work, and visual symptoms associated with microscope use were collected. All subjects had a comprehensive eye examination at The Hong Kong Polytechnic University Optometry Clinic, including measures of refractive error (both noncycloplegic and cycloplegic), binocular vision functions, and axial length. RESULTS: The prevalence of myopia in this group of microscopists was 87%, the mean (+/- SD) refractive error was -4.45 +/- 3.03 D and mean axial length was 25.13 +/- 1.52 mm. No correlation was found between refractive error and years spent working as a microscopist or number of hours per day spent performing microscopy. Subjects reporting myopia progression (N = 22) did not differ from the refractively stable group (N = 19) in terms of their microscopy working history, working hours, tonic accommodation level, or near phoria. However, the AC/A ratio of the progressing group was significantly greater than that of the stable group (4.59 delta/D cf. 3.34 delta/D). CONCLUSION: The myopia prevalence of Hong Kong Chinese microscopists was higher than that of microscopists in the United Kingdom (87% cf. 71%), as well as the Hong Kong general population (87% cf. 70%). The average amount of myopia was also higher in the Hong Kong Chinese microscopists than the Hong Kong general population (-4.45 D cf. -3.00 D). We have confirmed that the microscopy task may slightly exacerbate myopia development in Chinese people. PMID- 15127928 TI - The prevalence of astigmatism in Taiwan schoolchildren. AB - PURPOSE: To understand the prevalence and distribution of astigmatism in schoolchildren in Taiwan, we analyzed and compared the nationwide survey data in 1995 and 2000. METHODS: A total of 11,175 students were enrolled in 1995, and 10,878 students were enrolled in 2000. The refractive status of each student was measured with an autorefractor during cycloplegia and rechecked with retinoscopy. RESULTS: About half of schoolchildren (57.5% in 1995 and 49.0% in 2000) had no astigmatism (<0.5 D). About one third of schoolchildren's astigmatism was <1 D (27.9% vs. 32.6%). Eleven percent of schoolchildren in 1995 and 13% in 2000 had astigmatism between 1.0 and 2.0 D. Less than 2% of students had astigmatism >3.0 D (1.3% in 1995 and 1.8% in 2000). Most astigmatism was with-the-rule: 83.3% in 1995 and 89.9% in 2000. Only 16.6% of children in 1995 and 9.7% in 2000 had against-the-rule astigmatism. Very little astigmatism was oblique (0.1% in 1995 and 0.4% in 2000). The rate of myopic astigmatism increased with age. In contrast, the rate of hyperopic and mixed astigmatism decreased with age. In addition, the rate of with-the-rule astigmatism increased and the rate of against the-rule decreased with respect to age, but oblique astigmatism was rather stable with age. CONCLUSIONS: Most schoolchildren had little or no astigmatism. In Taiwan, most astigmatism is <1 D and is myopic with-the-rule astigmatism. There was more myopic astigmatism and with-the-rule astigmatism in 2000 than in 1995. PMID- 15127929 TI - Measurement of refractive state and deprivation myopia in two strains of mice. AB - PURPOSE: The mouse eye has a bright retinal image (f/number <1) but low optical quality (visual acuity about 0.5 cpd) that may render emmetropization unnecessary. However, this species is potentially a powerful model to study eye growth and myopia because its genome can be readily manipulated and has been completely sequenced. We have investigated how precisely eyes of mice can be refracted and tested whether deprivation myopia can be induced by frosted diffusers. METHODS: An automated eccentric infrared photorefractor was adapted to refract eyes of two mouse strains--C57BL/6 (B6) and DBA/2 (D2)--during Tropicamide cycloplegia without anesthesia. Axial lengths were measured in highly magnified video images of freshly excised eyes. Plastic hemispherical diffusers were applied between postnatal days and 29 and left attached for 7 or 14 days. RESULTS: (1) Trial lenses ranging from +10 to -10 D produced high correlations between the brightness slope in the pupil and applied lens power (r = 0.81 and r = 0.87), demonstrating reliable refraction. Five repeated measures in 12 eyes showed an average standard deviation of 3.0 D, equivalent to an axial length change <10 microm (derived from schematic eye modeling). (2) Deprivation produced a significant shift toward myopia, relative to untreated eyes, but only after 14 days and only in B6 mice (p = 0.02 with or p = 0.00038 without one outlier; N = 9). In contrast, DBA/2J were unaffected by occlusion, perhaps due to mutations that target eye, lens, or anterior segment. (3) Both eyes of untreated animals often had axial lengths that differed markedly. Surprisingly, we detected no significant correlation between refractive error and axial length after treatment. CONCLUSIONS: The infrared refraction technique is sufficiently sensitive to resolve equivalent changes in axial length of only +/- 10 microm in alert mice. Prolonged occlusion produces a significant myopic shift in B6 mice, but not in D2 mice. Even among isogenic B6 mice, the response is variable for reasons that presumably trace back to subtle developmental, environmental, and technical factors. PMID- 15127930 TI - The effect of the nonspecific nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester on the choroidal compensatory response to myopic defocus in chickens. AB - PURPOSE: Chick eyes show rapid compensation to retinal defocus. One component of this mechanism involves changes in the thickness of the choroid: when the retina is exposed to myopic defocus, the choroid thickens, pushing the retina forward; conversely, when the eye is exposed to hyperopic defocus, the choroid thins. The underlying mechanism(s) for these changes are unknown. We tested the hypothesis that nitric oxide might play a role. METHODS: We examined the effect of the nonspecific nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester (L NAME) on the compensatory choroidal thickening in response to myopic defocus using two visual paradigms: first, in previously form-deprived "recovering" eyes and, second, in eyes wearing +15 D spectacle lenses. L-NAME was injected intravitreally after removal of the diffuser or immediately before putting on the lenses. In addition, we looked at the effect of L-NAME on experimentally thickened choroids (induced by 1 week of recovery from deprivation myopia or 1 week of +15 D lens wear) and on choroids of normal eyes. Eyes were measured using A-scan ultrasonography before the injections and at subsequent intervals for several days. As a control for the injection procedure, eyes with the same visual conditions were injected with saline. Fellow eyes were untreated and uninjected. RESULTS: L-NAME inhibited choroidal thickening in both previously form-deprived eyes (2 vs. 117 microm; p < 0.001) and eyes wearing +15 D lenses (3 vs. 137 microm; p < 0.02). The effect was rapid, transient, and dose dependent (ED50, 0.26 micromoles). L-NAME produced thinning in experimentally thickened choroids (recovering: -116 microm; lenses: -219 microm) and in normal choroids (-47 microm) within 7 hours. CONCLUSIONS: Nitric oxide may play a role in modulating choroidal thickness. The mechanism is as yet unknown. PMID- 15127931 TI - Susceptibility to form-deprivation myopia in chicks is not altered by an early experience of axial myopia. AB - PURPOSE: Studies in humans and primates suggest that early visual experience may influence eye growth and refractive development later in life. In this study, we asked whether experimentally-induced myopia in 1-week-old chicks influences the responsiveness to form deprivation at a later age (4 weeks old). METHODS: A group of White Leghorn chicks ("twice deprived," N = 12) were monocularly deprived of form vision with white translucent diffusers at 3 days of age for 4 days. The diffusers were then removed, and the chicks were allowed 3 weeks of normal vision to age 27 days before being deprived again for 4 days. Another group of chicks ("once deprived," N = 9) were monocularly deprived of form vision at age 27 days for 4 days. Refractive errors, corneal curvatures, and axial ocular dimensions were measured by retinoscopy, infrared videokeratometry, and A-scan ultrasonography, respectively. Measurements were performed daily during the periods of deprivation and at approximately 3-day intervals in between treatments and after the final treatment period. RESULTS: The magnitude of the form deprivation myopia induced by 4 days of deprivation at 27 days of age was significantly smaller than that induced by the same treatment at 3 days of age ( 4.1 vs. -9.8 D; paired t-test, p < 0.01). This difference in induced myopia reflects optical scaling with increasing eye size because the deprivation-induced changes in vitreous chamber depth were not significantly different for the two deprivation periods (0.37 vs. 0.35 mm, paired t-test, p = 0.65). The induction of myopia at the younger age did not affect the susceptibility to form-deprivation myopia at the older age; there was no difference in the response to form deprivation at the older age between the once-deprived and twice-deprived groups (-3.5 vs. -4.1 D; unpaired t-test, p = 0.50). There was also a significant correlation between the amount of axial elongation induced in individual eyes during the first and second periods of deprivation (r = 0.631, p < 0.05). CONCLUSIONS: The induction of form-deprivation myopia at a young age does not affect the response to form deprivation at a later age. The significant correlation between the axial elongation induced in individual eyes over the two successive periods of deprivation suggests individual differences, possibly genetic in origin, in the susceptibility to form-deprivation myopia in chicks. PMID- 15127932 TI - Effects of quisqualic acid on retinal ZENK expression induced by imposed defocus in the chick eye. AB - PURPOSE: Expression of the transcription factor ZENK in glucagon amacrine cells of the chicken retina is enhanced after treatment with positive spectacle lenses and reduced after treatment with negative lenses. ZENK may, therefore, have an important role in emmetropization. To learn more about its regulation, we have studied its expression after retinal intoxication with quisqualic acid (QA, a glutamatergic excitotoxin). METHODS: Lenses of either +7 or -7 D power were placed in front of the eyes of young chickens 6 days after intravitreal QA injections. By this time, QA had caused severe damage to the retina. After 2 hours of lens wearing, changes in ZENK immunoreactivity were measured by means of double staining. In another experiment, lenses were worn for 4 days to study the residual function of emmetropization. RESULTS: QA injections caused a massive loss of cells in the inner nuclear layer and the ganglion cell layer but left the numbers of glucagon cells unchanged. Four of six QA-injected eyes became more myopic in response to wearing positive lenses, and all eyes with negative lenses also became myopic. QA caused a general reduction in ZENK expression, and there was no clear evidence that ZENK expression was still controlled by the sign of imposed defocus. CONCLUSIONS: After severe destruction of the inner retina by QA, retinal image processing appeared to be reduced to blur detection with no sign, causing myopia with both types of lenses. QA must remove synaptic input to the glucagon cells, which is necessary to transmit the information on the sign of imposed defocus. PMID- 15127933 TI - Inhibitory effects of apomorphine and atropine and their combination on myopia in chicks. AB - PURPOSE: The inhibitory effect of apomorphine on form-deprivation myopia implies a role for dopaminergic pathways in eye growth; however, the effect of apomorphine on lens-induced changes has not been studied. Our study filled this deficiency. After establishing that apomorphine inhibited lens-induced myopia, we investigated whether apomorphine and atropine acted sequentially via the same control pathway or via different parallel pathways. METHODS: This study, conducted in 8-day-old chicks, was comprised of two parts: (1) a comparative study of apomorphine's effect on lens-induced myopia (-15 D), form-deprivation myopia (diffusers), and lens-induced hyperopia (+15 D) and (2) a study of the interacting effects of apomorphine and atropine on lens-induced myopia and form deprivation myopia. In the first part, dH2O and six apomorphine doses (8 pmole to 800 nmole in log10 steps) were given as 10-microL intravitreal injections in combination with the above visual treatments. Apomorphine was used alone or given with atropine in the second part, which included four drug treatment groups: (1) control (dH2O); (2) 80 pmole of apomorphine; (3) 18 nmole of atropine; and (4) apomorphine + atropine. Additional dH2O injections were used to equalize the number of injections across groups. After 4.5 days of treatment, refractive errors and axial ocular dimensions were measured. RESULTS: The myopic shifts and axial elongation typical of lens-induced myopia (-15 D lens wear) were inhibited to maxima of 43% (4.5 D) and 52% (0.17 mm) by apomorphine, which, in contrast, enhanced lens-induced hyperopia (refractive error: 114%, 1.55 D; axial length: 134%, 0.16 mm). Inhibitory effects of apomorphine on lens-induced myopia were observed at doses > or = 80 pmole, whereas the doses required to enhance lens induced hyperopia were 2 log10 units higher. Only a weak inhibitory effect of apomorphine on form-deprivation myopia was observed. Although both apomorphine and atropine inhibited lens-induced myopia, atropine was slightly more effective for the doses compared (refractive error, 53% cf. 32%), and the effect of the combination was not significantly greater than that of atropine alone (refractive error, 59% cf. 53%). CONCLUSIONS: Apomorphine inhibits both types of experimental myopia, which implies the involvement of dopaminergic mechanisms in both phenomena; likewise, cholinergic mechanisms are indicated by the inhibitory effects of atropine on both lens-induced myopia and form-deprivation myopia. We speculate that apomorphine and atropine act at different sites on a common control pathway because the combined effect of apomorphine and atropine was no more than atropine alone. PMID- 15127934 TI - Screening for differential gene expression during the development of form deprivation myopia in the chicken. AB - PURPOSE: To use the technique of differential gene display to analyze changes in gene expression that occur during the development of and recovery from form deprivation myopia. METHODS: The differential display-reverse transcriptase polymerase chain reaction technique was used to detect cDNAs that are differentially expressed after 24 h (including 12 h in the light) after fitting with a diffuser to induce form-deprivation myopia. Messenger RNA levels were determined by quantitative Northern blotting in retinas after 11 days of form deprivation or in retinas where the diffusers had been removed the previous day. RESULTS: Twenty-six differentially expressed genes were processed in our initial screen. Two of these, alphaB-crystallin and retinoic acid receptor-alpha, were studied further. Levels of alphaB-crystallin mRNA were increased on day 11 in retinas from form-deprived eyes relative to eyes of control chickens and were reduced to below those levels within 6 to 12 h after removal of the diffusers. Levels of retinoic acid receptor-alpha mRNA showed similar changes, except that after removal of the diffusers, the levels further increased. CONCLUSIONS: The technique of differential gene display can be used to detect changes in gene expression during the regulation of eye growth. The response of alphaB-crystallin is particularly interesting because expression increases when eye growth is high and decreases when eye growth slows. PMID- 15127935 TI - Prediction of amino acid pairs sensitive to mutations in the spike protein from SARS related coronavirus. AB - In this study, we analyzed the amino acid pairs affected by mutations in two spike proteins from human coronavirus strains 229E and OC43 by means of random analysis in order to gain some insight into the possible mutations in the spike protein from SARS-CoV. The results demonstrate that the randomly unpredictable amino acid pairs are more sensitive to the mutations. The larger is the difference between actual and predicted frequencies, the higher is the chance of mutation occurring. The effect induced by mutations is to reduce the difference between actual and predicted frequencies. The amino acid pairs whose actual frequencies are larger than their predicted frequencies are more likely to be targeted by mutations, whereas the amino acid pairs whose actual frequencies are smaller than their predicted frequencies are more likely to be formed after mutations. These findings are identical to our several recent studies, i.e. the mutations represent a process of degeneration inducing human diseases. PMID- 15127936 TI - Novel Gly building units for backbone cyclization: synthesis and incorporation into model peptides. AB - We report the preparation of novel building units for backbone cyclization that have the general formula Fmoc-Nalpha[CH(R)CO2Al]Gly-OH. These building units were prepared by the reductive alkylation method using allyl esters of several amino acids as starting material and hence, respectively, contain the side chain of these amino acids. These N-alkylated Gly building units were incorporated in model backbone cyclic peptides. The resulting crude backbone cyclic peptides were obtained in high degree of purity according to HPLC and mass spectrometric analyses. PMID- 15127937 TI - Influence of peptide conformation on oligosaccharide binding characteristics--a study using apamin-based chimeric peptide. AB - Interactions between proteins and heparin play a crucial role in most of the cellular process. Unraveling the forces that govern the formation of these complexes is vital for understanding the specificities involved in these biomolecular events. In the present study, a detailed analysis has been undertaken to evaluate the effect(s) of peptide conformation on heparin-binding, using a chimeric peptide, apaK6--a chimera of a highly stable neurotoxic peptide from honey-bee venom and a de novo designed lysine-rich peptide. The dissociation constants of these peptide-heparin complexes were found to be in the submicromolar range. Comparison of the results obtained from the titration of the disulfide-reduced and disulfide-intact chimeric peptide with various sulfated oligosaccharides, derived from heparin, suggest that the initial structure of the peptide has pronounced effect on the binding affinity, binding modes and also on binding preferences. The results of this study indicate that the heparin-binding specificity of an isolated peptide and that exhibited by the same peptide when present in a globular protein could be significantly different, especially if the isolated peptide undergoes conformational change(s) upon binding to the sulfated oligosaccharides. In addition, such dependency of the binding specificity on the preformed structures could be utilized for the design of high-affinity and sequence-specific heparin-binding polypeptides. PMID- 15127938 TI - Correlations of amino acids in proteins. AB - A correlation analysis among 20 amino acids is performed for four protein structural classes (alpha, beta, alpha/beta, and alpha+beta) in a total of 204 proteins. The correlation relationships among amino acids can be classified into the following four types: (1) strong positive correlation, (2) strong negative correlation, (3) weak correlation, and (4) no correlation. The correlation relationships are different for different proteins and are correlated with the features of their structural classes. The amino acids with the weak correlation relationship can be treated as the independent basis functions for the space where proteins are defined. The amino acids with large correlation coefficients are linear correlative with each other and they are not independent. The strong correlation among amino acids reflects their mutual constrained relationship, as exhibited by their relevant structural features. The information obtained through the correlation analysis is used for predicting protein structural classes and a better prediction quality is obtained than that by the simple geometry distance methods without taking into account the correlation effects. PMID- 15127939 TI - Two genetic variants of the crustacean hyperglycemic hormone (CHH) from the Australian crayfish, Cherax destructor: detection of chiral isoforms due to posttranslational modification. AB - From sinus glands of the Australian crayfish Cherax destructor, two genetic variants of the crustacean hyperglycemic hormone (CHH) were isolated by HPLC and fully characterized by mass spectrometry and Edman sequencing. Both CHH A (8350.38 Da) and CHH B (8370.34 Da) consist of 72 amino acid residues, with pyroGlu as N-terminus and an amidated (Val-NH2) C-terminus. They differ in 14 residues (81% identity). Both sequences are significantly different from those of the hitherto known three CHHs of Astacoidea species (Northern hemisphere crayfish), which among themselves are extremely conserved. This may reflect the long, separate evolution of the Astacoidea lineage and the Parastacoidea (Southern hemisphere crayfish) lineage, to which Cherax belongs. CHH A and CHH B genes are expressed at comparable levels, as indicated by the similar amounts of mature peptides in the sinus gland. In addition to each of the major peptides, which share the identical N-terminal tripeptide pyroGlu-Val-L-Phe, one chiral isoform containing pyroGlu-Val-D-Phe was identified. Compared to the main peptides, the amounts of the D-isoforms are lower, but significant, amounting to 30-40% of L-isoforms. These results demonstrate that two genes can give rise to a total of four different peptides in the secretory terminals of the sinus gland. All peptides gave a highly significant hyperglycemic in vivo response in C. destructor. PMID- 15127940 TI - Conformational states of the corticotropin releasing factor 1 (CRF1) receptor: detection, and pharmacological evaluation by peptide ligands. AB - Previous corticotropin releasing factor 1 (CRF1) receptor characterization has been performed using radiolabeled agonists, which bind predominantly the receptor G-protein complex. The pharmacological profile of other receptor states, and their abundance, remain poorly characterized. Here we investigated the affinity states of the CRF1 receptor heterologously expressed in Ltk- cells and endogenously expressed in rat cerebellum. In L-CRF1 cell membranes, three agonist affinity states were detected: a very-high affinity receptor-G-protein complex state (eliminated by GTPgammaS) bound by [125I]sauvagine (43 pM, RG); a high affinity state insensitive to GTPgammaS bound by [125I]sauvagine (1.4 nM, termed RO); and a low affinity G-protein-uncoupled state detected by sauvagine displacement of [125I]astressin, a labeled antagonist (120 nM, R). The relative abundance of RG:RO:R was 18%:16%:66%. All three states were demonstrated in rat cerebellum with similar relative abundance (15%:16%:69%). The R state bound CRF with low affinity (270-330 nM), displayed a novel rank order of ligand affinity, and represented the majority of the receptor population in both receptor preparations. This study provides a framework to identify CRF1 receptor conformational states in various receptor preparations. PMID- 15127941 TI - Chimeric NDP-MSH and MTII melanocortin peptides with agouti-related protein (AGRP) Arg-Phe-Phe amino acids possess agonist melanocortin receptor activity. AB - Agouti-related protein (AGRP) is one of only two known endogenous antagonists of G-protein coupled receptors (GPCRs). Specifically, AGRP antagonizes the brain melanocortin-3 and -4 receptors involved in energy homeostasis, regulation of feeding behavior, and obesity. Alpha-melanocyte stimulating hormone (alpha-MSH) is one of the known endogenous agonists for these receptors. It has been hypothesized that the Arg-Phe-Phe (111-113) human AGRP amino acids may be mimicking the melanocortin agonist Phe-Arg-Trp (7-9) residue interactions with the melanocortin receptors that are important for both receptor molecular recognition and stimulation. To test this hypothesis, we generated thirteen chimeric peptide ligands based upon the melanocortin agonist peptides NDP-MSH (Ac Ser-Tyr-Ser-Nle4-Glu-His-DPhe-Arg-Trp-Gly-Lys-Pro-Val-NH2) and MTII (Ac-Nle-c[Asp His-DPhe-Arg-Trp-Lys]-NH2). In these chimeric ligands, the agonist DPhe-Arg-Trp amino acids were replaced by the AGRP Arg-Phe-Phe residues, and resulted in agonist activity at the mouse melanocortin receptors (mMC1R and mMC3-5Rs), supporting the hypothesis that the AGRP antagonist ligand Arg-Phe-Phe residues mimic the agonist Phe-Arg-Trp amino acids. Interestingly, the Ac-Ser-Tyr-Ser-Nle4 Glu-His-Arg-DPhe-Phe-Gly-Lys-Pro-Val-NH2 peptide possessed 7 nM mMC1R agonist potency, and is 850-fold selective for the mMC1R versus the mMC3R, 2300-fold selective for the mMC1R versus the mMC4R, and 60-fold selective for the MC1R versus the mMC5R, resulting in the discovery of a new peptide template for the design of melanocortin receptor selective ligands. PMID- 15127942 TI - Refractory hypothalamic alpha-mSH satiety and AGRP feeding systems in rats bearing MCA sarcomas. AB - In pre-anorectic tumor-bearing (TB: methylcholanthrene-induced sarcoma) rats, injection of alpha-melanocyte stimulating hormone (alpha-MSH) into the perifornical hypothalamus (PFH) had no significant effect on food intake at a dose (5 microg) that reduced feeding in non-TB control rats. Following the development of anorexia, injection of alpha-MSH MC3/MC4 receptor antagonists, SHU9119 (1 microg) or 4 microg agouti-related protein (AGRP), stimulated feeding in non-TB rats, while having no significant effect in TB rats. Concentrations of alpha-MSH were not altered significantly in ventromedial, dorsomedial or lateral hypothalamic areas of TB rats, and proopiomelanocortin (POMC) messenger RNA was not changed in TB rats in these hypothalamic areas. Determination of cytokines by ELISA in non-operated TB and non-TB rats revealed elevated IL-2 in plasma and hypothalamus as well as increased TNF-alpha in the hypothalamus of anorectic TB rats. IL-1B was not detectable in plasma and was not altered significantly in hypothalamus of TB rats. These results suggest that the POMC alpha-MSH satiety system is refractory in TB rats, even prior to the onset of anorexia. This change in MC3/MC4 receptor response does not appear to be secondary to alterations of endogenous alpha-MSH in TB rats. Cytokine involvement in the altered response to MC3/MC4 receptor stimulation and blockade is a possibility, since TNF-alpha and IL-2 were increased in hypothalamus of anorectic TB rats. Therefore, these results suggest major alterations in POMC neuropeptide systems in TB rats as anorexia progresses. Although these changes do not appear to have occurred due to grossly-altered concentrations of alpha-MSH, elevated cytokine activity in the hypothalamus may be an important factor. Due to the complex multi-factorial nature of feeding control, additional factors are likely to be involved in cancer anorexia. PMID- 15127943 TI - Synaptic interactions between ghrelin- and neuropeptide Y-containing neurons in the rat arcuate nucleus. AB - Morphological relationships between neuropeptide Y- (NPY) like and ghrelin-like immunoreactive neurons in the arcuate nucleus (ARC) were examined using light and electron microscopy techniques. At the light microscope level, both neuron types were found distributed in the ARC and could be observed making contact with each other. Using a preembedding double immunostaining technique, some NPY immunoreactive axon terminals were observed at the electron microscope level to make synapses on ghrelin-immunoreactive cell bodies and dendrites. While the axo somatic synapses were mostly symmetric in nature, the axo-dendritic synapses were both symmetric and asymmetric. In contrast, ghrelin-like immunoreactive (ghrelin LI) axon terminals were found to make synapses on NPY-like immunoreactive (NPY LI) dendrites although no NPY-like immunoreactive perikarya were identified receiving synapses from ghrelin-LI axon terminals. NPY-like axon terminals were also found making synapses on NPY-like neurons. Axo-axonic synapses were also identified between NPY- and ghrelin-like axon terminals. The present study shows that NPY- and ghrelin-LI neurons could influence each other by synaptic transmission through axo-somatic, axo-dendritic and even axo-axonic synapses, and suggests that they participate in a common effort to regulate the food-intake behavior through complex synaptic relationships. PMID- 15127944 TI - Adaptation to low-protein diet increases inhibition of gastric emptying by CCK. AB - Chronic nutritional disorders such as protein malnutrition are associated with delayed gastric emptying and increased postprandial cholecystokinin (CCK) levels. This study investigated the mechanisms involved in gastric emptying adaptation to low-protein diet. Two groups of 12 rats were adapted to a low-protein (LPD) or standard diet (SD) for 3 weeks. As compared to rats fed a SD, in rats adapted to a LPD gastric emptying was delayed, whereas postprandial CCK levels were increased. LPD enhanced antral muscle contractile response to CCK and cerulein without altering response to acetylcholine. This increased contractility was associated with up-regulation of CCK-A receptor mRNA levels in antral muscle. Our data suggest that modulation of gastric emptying after adaptation to a low protein diet involves up-regulation of both CCK-A receptors and CCK-induced contraction of antral smooth muscle. PMID- 15127945 TI - Effects of chronic ethanol on brain and serum level of methionine enkephalin. AB - Most evidence agrees that levels of methionine enkephalin (Met-Enk) in brain are inversely correlated with ethanol drinking and withdrawal seizures. One area of discrepancy is the effect of chronic ethanol administration on the level of immunoactive Met-Enk in brain, with some authors reporting increased and others reporting decreased levels. These reports differed greatly in terms of method of ethanol administration, species used, length of time ethanol was administered, and the region of brain examined. We found that all studies could be resolved by considering only length of time ethanol was administered, with Met-Enk levels first increasing and then decreasing. We tested this finding by determining the effect of 4-56 days of ethanol delivered in liquid feed on levels of brain Met Enk. We found that brain levels of Met-Enk peaked after 7 days of ethanol ingestion and declined to levels lower than control by 28 days. Exposure to ethanol abolished a correlation between brain and serum levels of Met-Enk which occurred in controls. HPLC showed that whereas 100% of immunoactivity eluted in the position of Met-Enk in controls, only about 50% eluted as Met-Enk in mice exposed to ethanol. These results support the hypothesis that exposure to ethanol alters brain Met-Enk in a way consistent with the reinforcement of physical dependence. PMID- 15127946 TI - Beta-endorphin-like peptide SLTCLVKGFY reduces the production of 11 oxycorticosteroids by rat adrenal cortex through nonopioid beta-endorphin receptors. AB - Beta-endorphin-like peptide immunorphin (SLTCLVKGFY), a selective agonist of nonopioid beta-endorphin receptor, was labeled with tritium to specific activity of 24 Ci/mmol. It was used for the detection and characterization of nonopioid beta-endorphin receptors on rat adrenal cortex membranes (Kd = 31.6 +/- 0.2 nM, Bmax = 37.4 +/- 2.2 pmol/mg protein). Immunorphin at concentrations of 10(-9) to 10(-6) M was found to inhibit the adenylate cyclase activity in adrenal cortex membranes, while intramuscular injection of immunorphin at doses of 10-100 microg/kg was found to reduce the secretion of 11-oxycorticosteroids from the adrenals to the bloodstream. PMID- 15127947 TI - Functional properties of Pfr(Tic)amide and BIBP3226 at human neuropeptide FF2 receptors. AB - The functional characteristics of two putative neuropeptide FF (NPFF) antagonists, BIBP3226 and PFR(Tic)amide, on the human neuropeptide FF receptor subtype 2 (hNPFF2) were investigated. Surprisingly, PFR(Tic)amide was shown to exhibit agonist properties in the [35S]guanosine-5'-O-(3-thio)triphosphate ([35S]GTPgammaS) binding assay. The efficacy of PFR(Tic)amide was significantly greater than that of (1DMe)Y8Fa, a stable analog of NPFF, and PFR(Tic)amide can therefore be classified as a 'super-agonist'. BIBP3226 did act as a reversible competitive antagonist on the hNPFF2 receptor. However, high concentrations of BIBP3226 also non-specifically increased [35S]GTP-gammaS binding. The usefulness of BIBP3226 as an antagonist tool on the NPFF receptor is thus limited. PMID- 15127948 TI - Beta-lactotensin and neurotensin rapidly reduce serum cholesterol via NT2 receptor. AB - Beta-lactotensin, a neurotensin NT2 agonist derived from beta-lactoglobulin, has hypocholesterolemic activity after administration for 2 days at a dose of 30 mg/kg (i.p.) or 100 mg/kg (p.o.) for 2 days in mice fed a high-cholesterol/cholic acid diet. The onset of hypocholesterolemic activity of beta-lactotensin was observed 90 min after a single i.p. or p.o. administration at the same dose as described above. Neurotensin also induced hypocholesterolemic activity 90 min after single i.p. administration at a dose of 2 microg per mouse but was ineffective after oral administration. The rapid onset of hypocholesterolemic activities of beta-lactotensin and neurotensin was blocked by levocabastine (50 microg/kg), an NT2 antagonist, and raclopride (0.5 mg/kg), a dopamine D2 antagonist. PMID- 15127949 TI - Impact of nitric oxide on adrenomedullin- and proadrenomedullin N-terminal 20 peptide-induced cardiac responses: action by alone and combined administration. AB - The cardiac effects of adrenomedullin (AM) and proadrenomedullin N-terminal 20 peptide (PAMP) as well as the possible signaling pathways were investigated. In the isolated perfused rat heart, infusion of AM (10(-11) to 10(-8) M) and PAMP(10(-11) to 10(-8) M) for 10 min, alone or in combination, induced concentration-dependent decreases in the left ventricular pressure (LVP), LVP +/- dp/dtmax of the hearts. The effects were attenuated by Nomega-nitro-L-arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) synthase. ADM and PAMP alone or in combinations increased the coronary fluid (CF), which could be antagonized by L-NAME. Pretreatment of H89, an inhibitor of protein kinase A (PKA), failed to alter the AM- or PAMP-induced decreases in LVP and LVP +/- dp/dtmax, but further promoted the AM or PAMP increased CF. The cAMP content in left cardiac ventricle was increased significantly by ADM infusions but not by PAMP. There was no statistical difference in cAMP contents with ADM administrated alone from those combined with ADM and PAMP. In conclusion, this study reveals that ADM and PAMP infused alone or in combinations inhibited the function of rat hearts in vitro, which may be partly involved with the NOS/NO pathway, rather than cAMP/PKA. PMID- 15127950 TI - Atrial natriuretic peptide and endothelin-3 target renal sodium-glucose cotransporter. AB - Atrial natriuretic peptide (ANP) and endothelin (ET) are endogenous vasoactive factors that exert potent diuretic and natriuretic actions. We have previously shown that ANP and ET-3 act through an NO pathway to inhibit the sodium-glucose cotransporter (SGLT) in the intestine [Gonzalez Bosc LV, Elustondo PA, Ortiz MC, Vidal NA. Effect of atrial natriuretic peptide on sodium-glucose cotransport in the rat small intestine. Peptides 1997; 18: 1491-5; Gonzalez Bosc LV, Majowicz MP, Ortiz MC, Vidal NA. Effects of endothelin-3 on intestinal ion transport. Peptides 2001; 22: 2069-75.]. Here we address the role of ANP and ET-3 on SGLT activity in renal proximal tubules. In rat renal cortical brush border membranes (BBV), fluorescein isothiocianate (FITC) labeling revealed a specific 72-kD peptide that exhibits increased FITC labeling in the presence of Na+ and D glucose. Using alpha-14C-methylglucose active uptake, rat BBV were shown to possess SGLT activity with an affinity constant (K(0.5) approximately 2.4 mM) that is consistent with the expression of the low-affinity, high-capacity SGLT2 isoform. SGLT2 activity in these preparations is dramatically inhibited by ANP and ET-3. This inhibition is independent of changes in membrane lipids and is mimicked by the cGMP analogue, 8-Br-cGMP, suggesting the involvement of cGMP/PKG pathways. These results are the first demonstration that both ANP and ET-3 inhibit rat cortical renal SGLT2 activity, and suggest a novel mechanism by which these vasoactive substances modulate hydro-saline balance at the proximal tubular nephron level. PMID- 15127951 TI - Human brain cathepsin H as a neuropeptide and bradykinin metabolizing enzyme. AB - Highly purified human brain cathepsin H (EC 3.4.22.16) was used to study its involvement in degradation of different brain peptides. Its action was determined to be selective. On Leu-enkephalin, dynorphin (1-6), dynorphin (1-13), alpha neoendorphin, and Lys-bradykinin, it showed a preferential aminopeptidase activity by cleaving off hydrophobic or basic amino acids. It showed no aminopeptidase activity on bradykinin, which has Pro adjacent to its N-terminal amino acid, on neurotensin with blocked N-terminal amino acid, or on dermorphin with second amino acid D-alanine. After prolonged incubation, cathepsin H acted as an endopeptidase. Dermorphin and dynorphin (1-13) were cleaved at bonds with Phe in the P2 position, while dynorphin (1-6), alpha-neoendorphin, bradykinin and Lys-bradykinin were cleaved at bonds with Gly in the P2 position. Further on, it was shown that human brain cathepsin H activity could be controlled in vivo by cystatin C in its full-length form or its [delta1-10] variant, already known to be co-localized in astrocytes, since the Ki values for the inhibition are in the 10(-10) M range. PMID- 15127952 TI - The role of calcium channels in substance P-induced contractile response in the rat iris. AB - This study was undertaken to assess the role of calcium channels in the contractile response induced by substance P in the isolated rat iris. Substance P produced graded and sustained contraction in the rat iris. Pre-incubation of preparations with thapsigargin (1 microM), verapamil (1 microM), isradipine (1 microM) or with omega-conotoxin MCIIA (0.1 microM) did not significantly inhibit substance P-mediated contraction in the isolated rat iris. However, pre incubation of the preparations with nicardipine (1 microM) or ruthenium red (1 mM) caused parallel displacement to the right of the substance P concentration response curve without affecting its maximal response. In contrast, amiloride (1 microM), markedly inhibited substance P-mediated contraction (73 +/- 5%), while econazole (1 mM) also significantly inhibited (44 +/- 11%) substance P-mediated contraction in the isolated rat iris. Collectively, these results suggest that substance P-mediated contractile response in the isolated rat iris depends largely on the influx of external Ca2+, by a mechanism which might involve the T type calcium channels. PMID- 15127953 TI - FDA aluminum rule poses challenges for industry, pharmacists. PMID- 15127954 TI - Army pharmacists' roles increase with mass rotation of troops. PMID- 15127955 TI - New drugs and dosage forms. PMID- 15127956 TI - Ephedrine alkaloids banned from dietary supplements. PMID- 15127957 TI - Omaha pharmacist helps city prepare for disasters. PMID- 15127958 TI - Wyoming program brings pharmacist consultations home. PMID- 15127959 TI - Appraisal of four novel approaches to the prevention and treatment of sepsis. AB - PURPOSE: Four novel approaches to the management of sepsis are discussed. SUMMARY: Drotrecogin alfa (activated) has FDA-approved labeling for use in the treatment of severe sepsis. Risk of bleeding and identification of the most suitable patients have been the major issues related to use of this drug. Tight glycemic control and early goal-directed therapy (EGDT) are promising supportive strategies. Both have challenged existing views regarding safe glucose levels and the usefulness of increased oxygen delivery in sepsis. The routine maintenance of euglycemia is resource intensive, however, and benefits during treatment of sepsis are unclear. Very early initiation of measures to optimize hemodynamic variables and the ability to identify patients with cryptic shock appear to be key reasons for successful EGDT. The use of corticosteroids for septic shock has been extensively researched and has provoked controversy. Selection of patients likely to benefit on the basis of relative adrenal insufficiency and prolonged treatment may account for recently observed positive results. A model for combining the four strategies is proposed. CONCLUSION: Novel strategies for treating sepsis include drotrecogin alfa (activated), tight glycemic control, EGDT, and low-dose corticosteroids. PMID- 15127960 TI - Hospital-based program for increasing the availability of emergency contraception: simulating nonprescription access. AB - PURPOSE: A hospital-based program simulating nonprescription access to emergency contraception (EC) is described. METHODS: A collaborative agreement between the pharmacy and therapeutics committee and the pharmacy department was initiated at a safety-net teaching hospital to provide EC to clinic patients directly from the hospital pharmacy without the need to first see a health care provider. EC was available 24 hours per day to any woman requesting it at the hospital pharmacy, with the collaborative agreement serving as the prescription. During clinic hours, patients were directed to the outpatient pharmacy to request EC. After hours, patients went to the emergency department triage desk and were directed to the inpatient pharmacy. Patients making inquiries about EC were encouraged to see their health care provider as soon as possible for counseling about contraceptive options. No specific program was initiated for publicizing the increased availability of EC, as it was assumed that health care providers and word-of mouth would inform patients of this option. RESULTS: The program was initiated in the fourth quarter of 2001. Total doses of EC dispensed increased nearly eightfold over the 1.5-year study period since the inception of this program. Most of this increase (81%) was attributable to the collaborative agreement. Twenty-eight percent of EC was dispensed outside of regular clinic hours. No patient complaints regarding this plan were received, and pharmacy staff did not believe that this program presented a significant additional burden to their workload. CONCLUSION: A collaborative agreement simulating nonprescription availability increased the use of EC in a hospital-based clinic setting. PMID- 15127961 TI - Antiemetic effectiveness of ondansetron and granisetron in patients with breast cancer treated with cyclophosphamide. AB - PURPOSE: The antiemetic effectiveness of ondansetron 8 mg i.v, ondansetron 32 mg i.v, and granisetron 10 microg/kg or 1 mg i.v. as prophylaxis in breast cancer patients regimens was studied. METHODS: Data from six U.S. cancer centers were collected retrospectively for 224 patients who received cyclophosphamide containing therapy between January 1998 and June 2002. Logistic-regression analysis was used to examine the likelihood of chemotherapy-induced nausea and vomiting (CINV) both on an unadjusted basis and controlling for concomitant radiation therapy and dexamethasone use. RESULTS: Seventy-six patients (34%) received ondansetron 32 mg, 68 (30%) received ondansetron 8 mg, and 80 (36%) received granisetron (either 10 microg/kg or 1 mg). Patients receiving ondansetron 8 mg were 2.5 times as likely to have CINV on an adjusted basis as granisetron recipients (p < 0.01). There was no increase in the risk of CINV with ondansetron 32 mg compared with granisetron. Patients treated with ondansetron 8 mg required more rescue antiemetics and more prophylactic antiemetics in subsequent chemotherapy cycles than patients in the other groups. CONCLUSION: In a retrospective multicenter study, granisetron 1 mg or 10 microg/kg and ondansetron 32 mg appeared more effective than ondansetron 8 mg in preventing acute CINV related to cyclophosphamide therapy. PMID- 15127962 TI - Pharmacist involvement in antimicrobial use at rural community hospitals in four Western states. AB - PURPOSE: Pharmacist involvement in antimicrobial use at small rural hospitals in four Western states was studied. METHODS: Surveys were mailed in July 2000 to hospitals with a daily patient census of <150 in Idaho, Nevada, Utah, and eastern Washington. RESULTS: Seventy-seven (77%) of 100 hospitals returned completed surveys. Only 5% of the hospitals had onsite pharmacists 24 hours per day. An onsite pharmacist was present for a median of 26 hours per week in hospitals without 24-hour pharmacist coverage (range, 0-116 hr/wk). Many hospitals (71%) had policies for monitoring or controlling antimicrobial use, but only 28% had a system capable of monitoring compliance with such policies. Few hospitals had systems for recommending changes in antimicrobial selection on the basis of susceptibility test results (27%) or for monitoring physician compliance with dosage recommendations by pharmacists (21%). Onsite pharmacist hours were significantly associated with pharmacists being involved in the initial ordering of antibiotics and providing active oversight of antimicrobial use. There was a negative correlation between onsite pharmacist hours and use of third-generation cephalosporins and carbapenems. CONCLUSION: A survey showed that rural hospital pharmacists in four Western states spent relatively little time monitoring and influencing antimicrobial prescribing. PMID- 15127963 TI - Pharmacy practice in Lebanon. PMID- 15127964 TI - Documentation of pharmacists' interventions and associated cost savings. PMID- 15127965 TI - A simple medication kit for emergency intubations. PMID- 15127966 TI - Hospitals should keep methylene blue. PMID- 15127967 TI - Counterfeit drugs: seeding the clouds? PMID- 15127968 TI - Tailoring immunosuppressive therapy in interstitial lung diseases. AB - An increasing number of immunosuppressive drugs have become available for clinical use over the past few years. Respiratory medicine has not been excluded from the growing enthusiasm devoted to the use of novel immunosuppressants. New agents are currently undergoing clinical trials in pulmonary disorders characterized by acute or chronic inflammation, the "optimal" immunosuppressive strategy for the cure of interstitial lung diseases in the next decade being forthcoming. An example of this fervour may be found in this issue of the Journal; data have been provided on the effectiveness in clinical use of an inhibitor of nucleotide synthesis, leflunomide, in chronic sarcoidosis. The great choice of available agents could allow us to select the best therapeutic regimen for an individual patient, however, this requires a comprehensive knowledge of the modes of action of the immunosuppressants we are planning to use. The present review provides an update of current understanding on the molecular mechanisms of some important immunosuppressants that are expected to play a role in the therapy of interstitial lung diseases. PMID- 15127969 TI - Functional genomics and prognosis in sarcoidosis--the critical role of antigen presentation. AB - BACKGROUND: Sarcoidosis is a systemic disorder of unknown cause, highly variable phenotype and unpredictable outcome. Antigen processing, inflammatory response and immunomodulation appear critical to development and prognosis of the disease. METHODS: We performed a comprehensive genomic analysis, applying high-density human GeneChip probe arrays (HUG95A, Affymetrix Inc.) for gene expression profiling from peripheral blood of patients with acute pulmonary sarcoidosis (n = 12) and matched healthy controls (n = 12), mean age 36 +/- 12 and 33 +/- 10 years respectively. RESULTS: At follow-up (18 [15-24] months), 7 patients had self limited disease and 5 had persistent disease. Significantly different expression comparing patients and controls was identified for 1,860 (14.9%) and 729 (5.8%) gene products at p = 0.05 and p = 0.01 levels respectively. Genes closely associated with persistent disease included HLA-DRB1*1501 DQB1*0602, TNFA, NFKB, cyclic AMP-responsive element modulator (CREM) and T-cell activation marker CD69. IL1B, IL8, growth related (GRO)-beta/-gamma and CCR 2,5,6 were closely associated with self-limited disease. CONCLUSION: We hypothesize that, in self-limited disease, greater effector cell activation leads to successful antigen elimination/tolerance, whereas HLA-DRB1*1501 DQBI*0602-mediated, probably defective/partial T-lymphocyte activation results in an inefficient primary immune response, antigen intolerance and persistent disease. PMID- 15127970 TI - Matrix metalloproteinase-1 polymorphism of promoter region in sarcoidosis and tuberculosis patients. AB - BACKGROUND AND AIM OF THE WORK: Matrix metalloproteinases (MMPs) are associated with several diseases. MMP-1 is one of the interstitial collagenases and the most highly expressed. Recent studies have found that a single nucleotide polymorphism located in the promoter region of the MMP-1 gene affects transcriptional activity. This polymorphism, 1G/2G, has been reported to associate with several malignant tumors and lung diseases. In this study, we investigated whether this polymorphism is associated with sarcoidosis or tuberculosis. METHODS: Polymerase chain reaction-restriction fragment length polymorphism was used to determine the MMP-1 genotypes of 103 sarcoidosis patients, 105 tuberculosis patients and 106 healthy control subjects. RESULTS: We found no differences in genotype distributions and allele frequency between sarcoidosis or tuberculosis patients and healthy control subjects. In sarcoidosis patients with ocular involvement, a significant increase in 1G/1G or 1G/2G genotype was observed compared with patients without (p = 0.009, odds ratio (OR) = 3.22, 95% confidence interval (CI): 1.34-7.71). In sarcoidosis patients with three or more organs involved, 1G/1G type tended to increase compared with patients without (p = 0.035, OR = 5.17, 95% CI: 1.12-23.9). In tuberculosis patients with cavity formation, an increasing trend of 1G/1G type was observed compared with patients without (p = 0.064, OR = 7.69, 95% CI: 0.89-66.3). CONCLUSIONS: Although MMP-1 polymorphism was not associated with onset risk of sarcoidosis and tuberculosis, the clinical characteristics of both diseases were affected by this polymorphism. PMID- 15127971 TI - Prolonged culture for mycobacteria in mediastinal lymph nodes from patients with pulmonary sarcoidosis. A negative study. AB - BACKGROUND: Histological similarity between sarcoidosis and tuberculosis, with epithelioid cell granulomas as the typical finding, suggests that sarcoidosis could be a specific manifestation of infection with mycobacteria. However, convincing evidence for a causal relationship between mycobacterial infection and sarcoidosis has not been produced. OBJECTIVE: To examine whether mycobacteria could be cultured from mediastinal lymph nodes from patients with newly diagnosed pulmonary sarcoidosis using a prolonged culture technique. METHODS: The series comprised 15 patients (10 men) with a median age of 45 years (range 25-61) and chest X-ray showing mediastinal lymphadenopathy. The diagnosis of sarcoidosis was based on biopsy specimens of lymph nodes obtained by mediastinoscopy. Lymph node tissue was processed in a prolonged (12 month duration) culture for mycobacteria. Bronchial washings were obtained by bronchoscopy and examined for mycobacteria using fluorescence microscopy, polymerase chain reaction (PCR) for Mycobacterium tuberculosis (M. tuberculosis) DNA and standard culture (7-week duration). RESULTS: In all lymph node specimens, prolonged culture for mycobacteria was negative. In all bronchial washings, microscopy for mycobacteria, analysis for M. tuberculosis DNA, and standard culture for mycobacteria was negative. CONCLUSION: Our study with prolonged culture failed to demonstrate the presence of mycobacteria in sarcoid tissue, especially M. tuberculosis and M. paratuberculosis, and does not suggest that mycobacterial infection could play a direct pathogenic role in patients with pulmonary sarcoidosis. PMID- 15127972 TI - Analysis of TNF-alpha promoter polymorphisms in the susceptibility to beryllium hypersensitivity. AB - BACKGROUND AND AIM OF THE WORK: In susceptible individuals beryllium (Be) exposure may cause Be-hypersensitivity (BH), leading to a spectrum of immune abnormalities ranging from the systemic responsiveness to Be to the CD4+ T-cell dominated chronic granulomatous pneumonitis known as berylliosis. Two gene markers have been previously associated with berylliosis: HLA-DP allelic variants carrying glutamate in position 69 of the beta-chain and the high TNF-alpha production-associated TNF-alpha promoter allele TNFA2. Since a number of TNF alpha promoter sequence variants have been associated with higher or lower gene expression, the entire TNF-alpha promoter region was screened for BH-associated variants. METHODS: Denaturating High Performance Liquid Chromatography (DHPLC) analysis followed by DNA sequence analysis of the heteroduplex observed was performed on a DNA bank obtained from a Be-exposed population composed of 73 subjects with BH and 43 Be-exposed controls. RESULTS: The data show that the TNF alpha TNFA2 variant (genotypic frequency: BH 26.7%, Be-exposed controls 5.8%; p < 0.0001), the -857T variant (BH 19.7%, Be-exposed controls 10.5%; p = 0.045) are significantly associated with BH and there is no linkage disequilibrium between them. Further, 64.4% of BH subjects carried at least one of higher TNF-alpha production-associated polymorphisms (Be-exposed controls 34.8%; p = 0.0036). CONCLUSIONS: The finding that TNF-alpha production-associated polymorphisms are carried at higher frequency by BH-affected compared to Be-exposed controls suggests that the TNF-alpha may play a central role in the determination of susceptibility to BH. PMID- 15127973 TI - Cyclooxygenase-2 is up-regulated in lung parenchyma of chronic obstructive pulmonary disease and down-regulated in idiopathic pulmonary fibrosis. AB - BACKGROUND AND AIM OF THE WORK: In vitro studies have suggested that fibroblasts from idiopathic pulmonary fibrosis (IPF) may have an impaired induction of cyclooxygenase (Cox)-2. We have investigated Cox-1 and Cox-2 expression in lung tissue from IPF. METHODS: Cox-1 and Cox-2 expression were determined using RT competitive PCR and immunohistochemistry in pulmonary biopsies from IPF (n = 22), chronic obstructive pulmonary disease (COPD) (n = 13), and lung tissue from subjects undergoing pleurodesis for spontaneous pneumothorax (control group, n = 17). RESULTS: Immunohistochemical analysis showed that the score of Cox-2 positive cells was higher in COPD (1 +/- 0) with respect to fibrosis (0.37 +/- 0.1, p < 0.05) and controls (0.57 +/- 0.2). There were no differences between fibrosis and controls in Cox-2 positive cells. The expression of Cox-2 mRNA was significantly higher in COPD (3.26 +/- 0.72 x 10(6) molecules cDNA/microg total RNA) in comparison to IPF (0.57 +/- 0.17) and controls (0.54 +/- 0.16) (p < 0.001). After IL-1beta stimulation (1-10 ng/ml) Cox-2 mRNA basal expression increased significantly in controls (from 35 +/- 12 to 94 +/- 4 x10(6) molecules cDNA/microg total RNA, p < 0.01) and in COPD (from 38 +/- 8 to 92 +/- 3, p < 0.01). In contrast, no significant changes in Cox-2 mRNA expression were found in IPF (from 30 +/- 12 to 43 +/- 16). CONCLUSIONS: Our results suggest that differences in Cox-2 expression may play a role in the regulation of inflammatory responses in lung diseases. Excessive activity is associated with the development of chronic obstructive lung disease, while a limited activation following pro inflammatory stimulation might contribute to fibrogenic responses. PMID- 15127974 TI - Leflunomide for chronic sarcoidosis. AB - BACKGROUND AND AIM: Leflunomide (Arava) is a cytotoxic drug which has been used as a single agent or in combination with methotrexate for the treatment of rheumatoid arthritis. It appears to have less toxicity than methotrexate. The use of leflunomide for sarcoidosis patients has not been systematically evaluated. METHODS: The records were reviewed from all patients treated at a tertiary sarcoidosis center from July 2002-July 2003, and information from patients treated with either leflunomide or methotrexate was analyzed for efficacy and toxicity. RESULTS: Thirty-two patients were treated with leflunomide; fifteen patients received concurrent therapy with methotrexate. The most common indications for therapy were ocular and lung disease. Complete or partial response to leflunomide was seen in 12 of 17 treated with leflunomide alone, and 13 of 15 treated with leflunomide plus methotrexate. There was no difference in the response rate for eye [23/28 (82%)] versus pulmonary disease [12/16 (75%)]. Seventeen patients were treated with leflunomide alone because of methotrexate toxicity (nausea in 12 and pulmonary symptoms in five). All but two tolerated leflunomide. Three patients experienced nausea leading to drug discontinuation, but no other serious adverse reaction was encountered with leflunomide. CONCLUSION: Leflunomide was well tolerated in patients with chronic sarcoidosis. It appears to be as effective as methotrexate, with less toxicity. It should be considered as an alternative in chronic sarcoidosis patients who cannot tolerate methotrexate. PMID- 15127976 TI - Relationship between perceived stress and sarcoidosis in a Dutch patient population. AB - BACKGROUND AND AIM: A relationship between stress and sarcoidosis has been considered. However, studies concerning perceived stress, appraisal of life events, are scarce in sarcoidosis patients. Therefore, the aim of the present study was to further examine the role of perceived stress in sarcoidosis. METHODS: Members of the Dutch Sarcoidosis Society (n = 1046; 59.0% females; the age range 40-49 contained the most persons) completed the Perceived Stress Scale (PSS), a symptom inventory, the Beck Depression Inventory (BDI), and the Fatigue Assessment Scale (FAS). RESULTS: The PSS score of sarcoidosis patients was high (p < 0.001), especially those of females (p < 0.001). Moreover, patients with psychological problems had higher PSS scores (p < 0.001). Notably, the presence of psychological problems and gender appeared to be unrelated. Furthermore, perceived stress was related to the BDI (r = 0.67, p < 0.001), especially to the cognitive subscale (r = 0.67, p < 0.001) and, to a lesser extent, to the physical depression subscale (r = 0.42, p < 0.001). CONCLUSIONS: Perceived stress was found to be high and related to symptoms in sarcoidosis. Moreover, depressive symptoms appeared to be related to perceived stress. Therefore, the management of sarcoidosis should include coping and appraisal therapy aiming to reduce stress and depressive symptoms. PMID- 15127975 TI - Photocopier exposure and risk of sarcoidosis in African-American sibs. AB - BACKGROUND: In a family study of sarcoidosis in African-Americans, we detected a positive association between sarcoidosis and ever working in a sales or clerical occupation. This finding, and case reports of granulomatous lung disease in patients with photocopier toner dust exposure, led us to hypothesize that sarcoidosis risk may increase as a result of photocopier exposure. METHODS: Retrospective data on photocopier use and maintenance were collected from African American sarcoidosis cases and their first degree relatives. The study sample consisted of 181 African-American sibships where one or more members had a history of sarcoidosis (n = 540). RESULTS: Sarcoidosis was statistically significantly associated with ever using a photocopier (Odds Ratio [OR] = 1.74, 95% confidence interval [CI]: 1.23-2.46), and ever changing photocopier toner or carrying out photocopier maintenance (OR = 2.88, 95% CI 1.83-4.54). In a conditional logistic regression model that adjusted for age and sex, the OR associated with a sarcoidosis history and being in the highest tertile of photocopier exposure ranged from 1.83 to 2.19 depending on the exposure measure used. CONCLUSIONS: These results suggest that photocopier toner dust may be a previously unrecognized antigen in the pathophysiology of some patients diagnosed with sarcoidosis. PMID- 15127977 TI - Report on the incidence of interstitial lung diseases in Spain. AB - BACKGROUND AND AIM OF THE WORK: Almost no epidemiological data are available on a worldwide basis on the prevalence, incidence or relative frequency of interstitial lung diseases (ILD). We report the results of a registration of ILD by 23 centers of pulmonary medicine in Spain over one year (from October 2000 to September 2001). METHODS: A standardized questionnaire was sent to the centers, together with guidelines for classification and diagnostic evaluation. This questionnaire included questions about the explorations performed to establish the diagnosis. RESULTS: A total of 511 cases were registered. The mean age of the patients was 61 +/- 0.7 (x +/- SEM) yrs. The male to female ratio was 1.2:1. The estimated incidence of ILD was 7.6 per 100,000/year. The most frequent disease was idiopathic pulmonary fibrosis (38.6%), followed in decreasing order by sarcoidosis (14.9%), cryptogenic organizing pneumonia (10.4%), ILD associated with collagen vascular diseases (9.9%) and hypersensitivity pneumonitis (6.6%). In 5.1% of cases ILD was unclassified. HRCT scan was performed in 91.9% of cases, bronchoalveolar lavage in 67.9%, transbronchial lung biopsy in 59.9%, and surgical lung biopsy in 22.7%. CONCLUSIONS: This registration provides interesting information on the occurrence of ILD in Spain and on the procedures used to establish the diagnosis. PMID- 15127978 TI - Cryptococcosis and sarcoidosis: strange bed-fellows. A report of five cases. AB - BACKGROUND AND AIM: Sarcoidosis is known to predispose to cryptococcal infection. In this case series, variations in presentation, diagnostic dilemmas and responses to treatment are highlighted. METHODS: Of several hundred patients referred to the sarcoidosis clinic at The Prince Charles Hospital, Brisbane and to the private clinic of the co-author between 1990 and 2002, five subsequently developed cryptococcal infections as a complication of sarcoidosis. All five subjects were treated by the co-author (RKAA). A review of the literature was also performed. RESULTS: Cryptococcal infection occurred in 4 patients with sarcoidosis being treated with steroids and one patient who was not on treatment. All responded to antifungal therapy. Cryptococcosis was diagnosed by transbronchial (2), bronchial (1) and cutaneous (2) biopsies. Fluconazole was used in those with pulmonary infections but not in cutaneous disease where excision sufficed. One patient with pulmonary cryptococcosis from immunosuppressants also developed a nocardial brain abscess. CONCLUSIONS: Our series of patients with sarcoidosis and cryptococcal infections is unique in Australia. Although an unusual infection, cryptococcosis should always be considered in patients with sarcoidosis as it may be overlooked particularly in the lungs and can be fatal if untreated. Further immune dysregulation through steroid use may contribute significantly to the disease manifestations. Understanding why cryptococcosis and not other infections is more common in patients with sarcoidosis may reveal more about the mechanisms of granuloma formation and the nature of sarcoidosis itself. PMID- 15127979 TI - The synovial fluid analysis in sarcoid arthritis. PMID- 15127980 TI - Novel aberrations of HPS1 mRNA detected in a case of Hermansky-Pudlak syndrome with pulmonary fibrosis. PMID- 15127981 TI - DO relates positive experience with use of integrated neuromusculoskeletal release. PMID- 15127982 TI - Musculoskeletal disorders: does the osteopathic medical profession demonstrate its unique and distinctive characteristics? AB - The authors used the National Ambulatory Medical Care Survey: 1999 Summary to compare the practice patterns of osteopathic and allopathic physicians in the management of musculoskeletal disorders in family practice settings. Patient and physician characteristics, diagnostic test ordering patterns, treatments, and amount of time spent with patients during visits were compared. Patients who visited osteopathic physicians were more likely to be middle-aged and referred, with injury-related visits that were self-paid. Osteopathic physicians spent more time with patients, ordered a greater number of nontraditional diagnostic tests, and provided more manual and complementary modes of therapy. In contrast, although most of the patients seen by both osteopathic and allopathic physicians were white, allopathic physicians had a greater percentage of patients who were of an ethnic minority or under Medicaid or Medicare. Allopathic physicians ordered a greater number of traditional diagnostic tests and prescribed more medications. Based on the nationally representative data, osteopathic physicians used physiotherapy (including osteopathic manipulative treatment and physical modes of therapy) and complementary treatments to a greater degree in their physician-patient contacts. In contrast, allopathic physicians spent more resources on diagnosis versus treatment (eg, physiotherapy) and seemed to focus on the search for a nonstructural medical cause. PMID- 15127983 TI - Early diagnosis is key in vancomycin-induced linear IgA bullous dermatosis and Stevens-Johnson syndrome. AB - BACKGROUND: With the emergence of highly resistant beta-lactam gram-positive organisms, vancomycin hydrochloride usage has increased considerably. Consequently, adverse drug reactions, including unfavorable cutaneous events, have also increased. Important adverse skin reactions are linear IgA bullous dermatosis (LABD) and Stevens-Johnson syndrome (SJS). These blistering disorders can have clinical manifestations that are difficult to distinguish; however, it is important to make the distinction because treatment and prognosis are different. OBJECTIVE: The purpose of this study is to review the literature on LABD and SJS and compare important differentiating characteristics to assist physicians in making the correct diagnosis. METHODS: The authors used MEDLINE to search for all published studies on vancomycin adverse events, and combined LABD, SJS, exanthema, and skin rashes each separately with vancomycin adverse events. Furthermore, the authors searched PubMed for all meta-analyses and randomized controlled clinical trials relating to treatment of patients with SJS. RESULTS: Clinically, LABD and SJS both present similarly with bullae. Diagnosis is made by use of perilesional skin biopsy and direct immunofluorescence. Direct immunofluorescence shows linear IgA deposition along the basement membrane zone in LABD, whereas this is absent in SJS. The treatment for both vancomycin-induced SJS and vancomycin-induced LABD is prompt discontinuation of the drug. However, if SJS is diagnosed early, systemic corticosteroids appear to decrease morbidity. CONCLUSIONS: In cases of SJS or LABD that are difficult to distinguish clinically, the authors recommend performing a skin biopsy and direct immunofluorescence early to confirm the diagnosis so that effective treatment can be instituted. PMID- 15127985 TI - A divisional approach to enhancing research among osteopathic family practice residents. AB - There is an increasing demand to expose osteopathic family practice residents to research. Within the Department of Family Medicine at the University of North Texas Health Science Center at Fort Worth-Texas College of Osteopathic Medicine, the Division of Education and Research (DEAR) was developed in 1999. The primary focus of DEAR is to provide the necessary resources and knowledge for faculty members and residents to conduct research and accomplish scholarly activities. Guidelines are implemented to ensure the timeliness, efficiency, and completion of each resident's paper. The efficacy of DEAR's policies is evaluated annually through surveying the residents, tracking paper submissions to the resident director, and tracking publications and presentations. The expectation of DEAR's resident research component is full completion of a resident paper suitable for publication in a peer-reviewed journal before graduation. Initial evaluation suggests positive strides toward reaching this goal. Future successes will be determined by monitoring the next few years. PMID- 15127984 TI - Common scenarios to clarify the interpretation of cardiac markers. AB - The authors present a practical approach for physicians in clinical practice to use cardiac troponins in the interpretation of heart disease and myocardial damage. Laboratory results that fall within the intermediate area of facility specific cutoff reference values for elevated troponin levels confer lower risks to patients than do higher levels of cardiac troponin. Perhaps not surprisingly, the actual anatomy of the vessels at cardiac catheterization does not correlate well with the troponin level. In the six cases presented here, the patients' low levels of troponin release are discussed using the new term minimal myocardial infarction, which is synonymous with conditions that would previously have been diagnosed as unstable angina. Elevated levels of cardiac troponin provide a very sensitive measure for clinicians diagnosing patients with myocardial necrosis, but such measures are also useful in defining a broad spectrum of disease. Whenever the troponin levels are elevated (barring laboratory error), the patient has a poorer prognosis. The greatest challenge for physicians is in determining which patients with cardiac troponin elevation will best benefit from heart catheterization and percutaneous intervention. PMID- 15127986 TI - [Intrauniversitary selection: a good or a bad way to identify the best medical students?]. AB - During their first two years at the medical faculty of the University of Lausanne, selection of medical students is based on their performance in tests in basic sciences. However, this carries the risk that scientifically talented students who perhaps do not necessarily possess the personal skills to make them a good doctor are given preferential admission to the clinical semesters. This study followed a student cohort (n = 115), which had passed the first part of their state examination at the end of the third year course. The examination included an oral examination in psychosocial medicine (PSM) that tested the student's skills with regard to patient-physician communication. The PSM grade is compared with the grade achieved in the written tests in physics and physiology taken at the end of the first year of study. The findings showed that the performance achieved in the basic science examination had no predicative value for the student's future performance in the PSM test. Moreover, it was found that the type of pre-graduate degree had no major influence on the student's progress in their preclinical years of medical school. PMID- 15127987 TI - [Highlights in gastroenterology 2003]. PMID- 15127988 TI - [Emergency admission of a 65-year-old patient with left neck pain and dysphagia for liquid food]. PMID- 15127989 TI - [Complementary medicine. Phytotherapy in gastrointestinal ulcer]. PMID- 15127990 TI - [History of Medicine. "Through the horny or ivory gate" -- dream interpretation in the Antiquity. I. From Homer to Aristotle]. PMID- 15127991 TI - Tapered implants in jaws with soft bone quality: a clinical and radiographic 1 year study of the Branemark System Mark IV fixture. AB - BACKGROUND: The survival rate of oral implants in soft-quality bone has been demonstrated to be inferior to that of implants inserted in good-quality bone. A possible way to increase the survival rate in soft-quality bone may be to use a tapered implant. Such an implant has been developed and manufactured by Nobel Biocare AB, Gothenburg, Sweden. So far, there have been only a few publications regarding this implant. PURPOSE: The aim of the study was to compare the outcome of using the tapered Branemark System Mark IV fixture with the outcome of using earlier Branemark fixtures in a controlled prospective study. MATERIALS AND METHODS: The study was performed as a multicenter study including seven specialist centers. The material consisted of 40 patients in need of implant supported bridges in the maxilla. Twenty-five patients belonged to the test group, in which the tapered Mark IV implants were inserted, and 15 patients belonged to the control group, in which Branemark Standard or Mark II implants were used. The patients were allocated to the test group or the control group according to randomization schedules. The implants were inserted according to the guidelines for Branemark implants. A two-stage surgical protocol was used, and abutment connection was made 6 months after fixture insertion. The test group comprised 97 Mark IV implants, and the control group made up 92 implants. The prosthetic procedure followed the guidelines for Branemark implants, and all patients were provided with full fixed maxillary bridges. The patients were followed up with clinical and radiographic records for 1 year after loading. RESULTS: The survival rate was 96.9% for the Mark IV implants and 98.9% for the control implants. There was no significant difference between the two groups. There was a mean marginal bone loss of 0.2 mm during the observation period, and there was no difference between test implants and control implants. CONCLUSIONS: With regard to survival rate and marginal bone level changes, no differences could be demonstrated between the Mark IV tapered implant and the Branemark implants used earlier. However, compared with earlier results of Branemark implants in soft-quality bone, the Mark IV implant demonstrated an improved survival rate. PMID- 15127992 TI - Clinical assessment and surgical implications of anatomic challenges in the anterior mandible. AB - BACKGROUND: The anterior mandible is generally considered a rather safe surgical area, involving few risks of damage to vital anatomic structures. Nevertheless, both neurosensory disturbances and hemorrhages have been reported after implant surgery in that particular area. PURPOSE: With the increasing demand for oral implant placement, the anatomy of the anterior mandible should receive more attention. This review will focus on the anatomic peculiarities of the anterior mandible and the related clinical implications. METHODS: The scientific evidence on the anatomic, histologic, physiologic, and clinical aspects of the neurovascularization of the anterior mandible will be reviewed. RESULTS: Surgical complications may be attributed to the existence of a mandibular incisive canal with a true neurovascular supply. Potential risks may also be related to the presence of the lingual foramen and anatomic variations, such as an anterior looping of the mental nerve. CONCLUSIONS: Preoperative radiographic planning for oral implant placement in the anterior mandible should therefore not only consider all esthetic and functional demands but should also pay particular attention to the anatomic peculiarities of this region to avoid any neurovascular complications. PMID- 15127993 TI - Microbiologic diagnostics at titanium implants. AB - BACKGROUND: The microbiota found at periimplant lesions have been shown to contain putative periodontal pathogens as well as opportunistic species such as Staphylococcus spp, enterics, and Candida spp. Therefore, a microbiologic diagnosis may be of value as guidance before treatment of such lesions. PURPOSE: The aim of this study was to evaluate the prevalence of some putative pathogens associated with long-term followed-up cases using two different microbiologic procedures. MATERIALS AND METHODS: Fifteen subjects contributed with plaque samples from teeth and implants; these were analyzed with respect to 18 putative periimplant pathogens using cultural methods and a deoxyribonucleic acid DNA-DNA hybridization technique. RESULTS: The number of individuals positive for the analyzed pathogens was similar in samples taken from teeth and implants when analyzed with the DNA-DNA hybridization technique. When comparing detection frequency by culture procedure and by "checkerboard" technique at implants, the number of individuals positive for these species was lower with the traditional culture technique than with the checkerboard analyses. Using a higher cutoff point (> or = 4) with the checkerboard technique, the number of positive individuals was generally lower than that found with the culture technique. When comparing the techniques on an implant site level, the prevalence obtained by culture was lower for all analyzed species. If the specific species were present in the samples analyzed by the checkerboard technique, they were present only in every second sample analyzed with the culture technique. The high specificity values showed that if the checkerboard technique did not detect any Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, or Fusobacterium nucleatum, the bacteria were also undetectable by the culture technique. The two methods therefore did not overlap but did supplement each other. CONCLUSIONS: Based on the current results it is recommended that the technique used when analyzing microbiota around titanium implants should be a combination of the two protocols mentioned as they seem to give the most comprehensive outcome when used together. PMID- 15127994 TI - Implant treatment in combination with lateral augmentation of the alveolar process: a 3-year prospective study. AB - BACKGROUND: In patients in whom the height of the alveolar process is adequate but the crest is too narrow to host an implant, lateral augmentation is required. Such augmentations have mostly been performed using autogenous bone blocks secured to the buccal surface. An alternative to autogenous bone may be bovine hydroxyapatite (Bio-Oss, Geistlich Pharma AG, Wolhusen, Switzerland) or other bone substitutes. PURPOSE: The aim of this study was to evaluate the clinical and radiographic outcome of dental implants inserted after lateral augmentation of too narrow alveolar processes with a combination of bovine hydroxyapatite (Bio Oss) and autogenous bone. METHODS: Thirty patients (14 males and 16 females) with a mean age of 41.6 years fulfilled the inclusion criteria. Twenty-nine augmentation sites with a total of 74 implants could be followed for 3 years. RESULTS: Three implants were lost; these were lost before loading (at the abutment operation). The survival rate was 95.9%. The mean marginal bone loss during the 3-year observation period was 0.3 +/- 0.2 mm. CONCLUSIONS: A 50/50 combination of Bio-Oss and autogenous bone chips stabilized with Tisseel (Baxter AG/Duo Quick AG, Vienna, Austria) was useful for lateral augmentation of the alveolar crest. Lateral grafts with Bio-Oss, autogenous bone, and Tisseel made it possible to achieve good implant stability and high implant survival results. The bone level changes adjacent to the implants were the same as in nongrafted cases. PMID- 15127996 TI - Influence of implant length and bicortical anchorage on implant stress distribution. AB - BACKGROUND: Short implants present superior failure rates for everybody. PURPOSE: The aim of this theoretic study was to assess to what extent implant length and bicortical anchorage affect the way stress is transferred to implant components, the implant proper, and the surrounding bone. MATERIALS AND METHODS: Stress analysis was performed using finite element analysis. A three-dimensional linear elastic model was generated. All implants modeled were of the same diameter (3.75 mm) but varied in length, at 6, 7, 8, 9, 10, 11, and 12 mm (Branemark System, Nobel Biocare AB, Gothenburg, Sweden). Each implant was modeled with a titanium abutment screw and abutment, a gold cylinder and prosthetic screw, and a ceramic crown. The implants were seated in a supporting bone structure consisting of cortical and cancellous bone. An occlusal load of 100 N was applied at a 30 degrees angle to the buccolingual plane. RESULTS: With the selected model and bone properties, the coronal cortical anchorage was dominating, and the bone stress concentrated to that area. CONCLUSIONS: The maximum bone stress was virtually constant, independent of implant length and bicortical anchorage. The maximum implant stress, however, increased somewhat with implant length and bicortical anchorage. PMID- 15127995 TI - Short-term bone response to titanium implants coated with thin radiofrequent magnetron-sputtered hydroxyapatite in rabbits. AB - BACKGROUND: It has been suggested that calcium phosphate (CaP) coatings initiate faster bone growth around implants. A major concern about the viable use of these coatings has been their biologic performance related to the coating characteristics. PURPOSE: The purpose of this study was to investigate the early bone response to micron- and submicron-thick hydroxyapatite (HA) coatings in cortical and trabecular bone. MATERIALS AND METHODS: CaP coatings were manufactured by magnetron sputtering. Heat treatment was subsequently used to increase the crystallinity of the coatings. Coatings were characterized by x-ray diffraction, Fourier transform infrared spectroscopy, inductively coupled plasma optical emission spectroscopy (ICP-OES), and stylus profilometry. Four types of CaP-coated implants were used (0.1 microm and 2.0 microm amorphous; 0.1 microm and 2.0 microm crystalline); uncoated machined commercially pure titanium implants served as controls. Four hundred eighty implants were randomly placed in 60 rabbits. Ten animals were followed up for 1 week, 10 for 3 weeks, and 40 for 6 weeks. The bone response was histomorphometrically evaluated. RESULTS: Coatings with a CaP ratio very close to that of HA were produced. Crystalline coatings significantly improved the early bone-implant contact whereas the amorphous coated implants behaved similarly to uncoated titanium. CONCLUSIONS: Crystalline CaP coatings 100 nm thick on titanium implants elicited an improved early bone response compared with that of uncoated titanium implants. No further improvement in the bone response was observed with 2 microm coatings. PMID- 15127997 TI - Surgical intervention in enchondral and membranous bone: intraindividual comparisons in the rabbit. AB - BACKGROUND: The long bones, which are of enchondral origin, are often used in histologic evaluations of dental implants that clinically will be placed in bones of membranous origin. PURPOSE: The aim of this investigation was to study the effect of surgical trauma by multiple drill perforations on both enchondral and membranous bone--the tibial and maxillary bone architecture--8 weeks after surgery. MATERIALS AND METHODS: In eight rabbits the medial metaphyses of both tibiae and the edentulous ridges of both sides in the maxilla were surgically exposed. On the test sides, 25 to 30 holes were drilled through the cortical plate and 5 mm into the medullary space. The contralateral sides underwent no treatment and served as controls. Eight weeks later the animals were killed to obtain transverse ground sections for histomorphometry. A paired analysis of the bone densities of the tibiae and the jaw specimens was made. RESULTS: In general the morphologic appearances of the test and control tibiae were similar. In the control group the anatomy of the medullary space appeared normal, with few bone trabeculae and with abundant marrow tissue. In the test group, areas of resorption were evident on the cortical bone plate that had been perforated 8 weeks previously; also evident were ingrowths of marrow tissue and bone formation. In a few specimens some newly formed bone trabeculae were found in the medullary space, both contiguous to the perforated cortical bone plate and as small islets in the marrow tissue. The mean test and control site values for each parameter were similar, and no statistically significant differences were found. In the maxilla, the bone architecture had altered substantially by way of an increase in the proportion of trabecular bone and a decrease in the proportion of cortical bone. Drilling appeared to have little effect on tibial bone density, which did not significantly differ between test and control sites. Maxillary bone density, however, was significantly greater at the test sites than at the control sites. Comparisons of bone density between tibia and jaw specimens revealed no significant differences either between test sides or between control sides. Thus, the same magnitude of surgical trauma that altered the bone architecture and density of the rabbit maxilla was not found to have altered those of the tibia in the same animal 8 weeks after surgery. The reasons for the differences are discussed in this article. CONCLUSIONS: As observed 8 weeks post surgery, multiple drill perforations were not found to cause significant alterations in the metaphysis of the tibia whereas a considerable change in bone architecture and an increase in bone density were found in the maxilla. Whether this depends on different embryologic origins or reflects morphologic and functional differences in the mature bone demands further study. PMID- 15127998 TI - Immediately loaded bar-connected implants with an anodized surface inserted in the anterior mandible in a patient treated with diphosphonates for osteoporosis: a case report with a 12-month follow-up. AB - BACKGROUND: It has been suggested that tooth loss is greater in the osteoporotic patient population. Only a few cases have been reported in the literature about the use of dental implants in patients with osteoporosis. Diphosphonates are stable analogs of pyrophosphate, a physiologic regulator of calcification and bone resorption. Multiple implant failures have been reported in a patient undergoing treatment with diphosphonates. Recently, several clinical and experimental reports have shown that immediate loading of dental implants is possible in selected situations. PURPOSE: The aim of this case report was to present the clinical outcome of immediate loading of implants in a patient undergoing diphosphonate treatment for osteoporosis. MATERIALS AND METHODS: In a 65-year-old patient undergoing diphosphonate treatment for osteoporosis, four implants were inserted in the anterior mandible. The implants were connected with a bar supporting an overdenture and were then loaded the same day. RESULTS: No problems occurred in the postoperative period. At 1-year follow-up, all four of the implants appeared to be clinically osseointegrated, and no mobility was present. Minimal bone resorption was present around all implants. CONCLUSIONS: Our case report points to the fact that, contrary to what has been reported in the literature, it is possible to successfully insert and load immediately after surgery dental implants in a patient undergoing diphosphonate treatment for osteoporosis. PMID- 15128000 TI - Uterine sarcomas: have we made any progress? PMID- 15127999 TI - Mk III: a third generation of the self-tapping Branemark System implant, including the new Stargrip internal grip design. A 1-year prospective four-center study. AB - BACKGROUND: Many prospective studies have shown predictable long-term results of the Branemark System (Nobel Biocare AB, Gothenburg, Sweden) turned-surface implants in totally edentulous, partially edentulous, and single cases. The Mk III self-tapping implant was introduced by Nobel Biocare in 1999 as part of a new implant generation that improved on the Mk II self-tapping implant that had been introduced in 1992. A new implant design should always be evaluated, preferably in prospective multicenter studies. PURPOSE: The aim of the present prospective multicenter study was to evaluate the short-term success rate of the Branemark Mk III machined- and turned-surface self-tapping implant and to evaluate the clinical handling of the new Stargrip (Nobel Biocare AB) internal grip design. MATERIALS AND METHODS: The open prospective 1-year multicenter study was performed at four clinics. Sixty-four patients (38 females, 26 males) with a mean age of 51 years (range, 16 to 80 years) were included. Seventeen of the patients were totally edentulous, 26 were partially edentulous, and 21 had single-tooth losses. In total, 194 Mk III implants were inserted, 89 in the maxillas and 105 in the mandibles. RESULTS: Fifty-eight patients were observed for 1 year after prosthetic loading. Three of 194 implants were lost, corresponding to a cumulative success rate of 98.4% after 1 year, 100% for maxillary implants and 97.1% for mandibular implants. The implant driver carried the implants reliably in 99.5% of all treated patients. Sixty-two patients received the planned prosthetic reconstruction. All but one prosthetic reconstruction survived after 1 year of loading. The mean marginal bone resorption, analyzed from radiographs of 143 implants, was 1.15 mm (standard deviation [SD], 1.09 mm) from the second stage surgery to the 1-year follow-up. CONCLUSION: The 1-year success rate of the Mk III implant was high, and the clinical handling of the internal grip design (Stargrip) improved the surgical technique and was well appreciated by both surgeons and assisting staff. PMID- 15128001 TI - New modalities in detection of recurrent ovarian cancer. AB - PURPOSE OF REVIEW: The vast majority of women diagnosed with ovarian cancer and subsequently treated with debulking surgery and adjuvant chemotherapy will ultimately relapse. As is the case with primary diagnosis, detection of recurrent ovarian cancer is limited due to lack of sensitivity and specificity. Specific guidelines for surveillance of this disease are controversial, partly because evidence to support such guidelines is scant and partly because the management of identified recurrences continues to be of minimal success. Subsequently, whether early detection actually can make a difference is not necessarily made clear in the literature. However, there are advances in radiological and molecular biology technology that may offer new possibilities in cancer surveillance. This review will outline the latest evidence to address their use in ovarian cancer. RECENT FINDINGS: Most of the recent literature involving detection of recurrent ovarian cancer addresses the use of positron emission tomography. There are also some data addressing the use of magnetic resonance imaging and computed tomography in this arena. Data pertaining to other modalities such as biological markers are limited. Ca-125 is the accepted assay used for ovarian cancer surveillance, but other options are introduced that may hold promise for the future. SUMMARY: A review of the recent literature concerning ovarian cancer surveillance techniques offers few new definitive avenues. While radiological technology and discoveries in detection assays are noteworthy, their potential impact on surveillance appears to be minimal at this time. Low sensitivity and specificity, along with expense, continue to be limiting factors. PMID- 15128002 TI - Surgical and radiographic staging in patients with cervical cancer. AB - PURPOSE OF REVIEW: The most recent data on surgical and radiographic staging in patients with cervical cancer are analysed. RECENT FINDINGS: Laparoscopic staging of retro and intraperitoneal disease is feasible. Morbidity is low, but the impact on survival has still to be shown. The sentinel lymph node concept is valid in patients with cervical cancer. Sensitivity and negative predictive value, however, have to be improved before the concept can be integrated into clinical practice. The majority of most recent radiographic studies evaluate the value of positron emission tomography. Accuracy of detection of extracervical disease is high and the response to chemoradiation can be measured by positron emission tomography using various scoring systems. Positron emission tomography seems to be the most accurate technique for detecting early recurrence. SUMMARY: Surgical staging is less invasive by laparoscopy, and radiographic staging becomes more accurate by positron emission tomography. Prospective evaluations have to show the impact of these new techniques on survival of patients diagnosed with cervical cancer. PMID- 15128003 TI - Role of vitamins in the risk, prevention, and treatment of breast cancer. AB - PURPOSE OF REVIEW: This article presents an overview of the role of vitamins in the risk, prevention, and treatment of breast cancer, with emphasis on current evidence from English-language articles published since 1 August 2002 and indexed in MEDLINE. RECENT FINDINGS: Findings from epidemiologic studies that have evaluated vitamin A and carotenoids in relation to breast cancer risk have been inconclusive. The available data, especially from prospective studies, do not support an association between vitamins E and C and risk of breast cancer. Recent studies suggest that folate plays an important role in the prevention of breast cancer, particularly among women consuming alcohol. Limited data also support a potential role of vitamin D in the prevention of breast cancer. SUMMARY: High intake of folate or adequate circulating levels of folate may reduce the risk of breast cancer. Adequate folate levels may be particularly important for women who are at higher risk of breast cancer because of high alcohol consumption. The inverse association between vitamin D and risk of breast cancer needs to be evaluated in more studies. PMID- 15128004 TI - Combined oral contraceptives and cervical cancer. AB - PURPOSE OF REVIEW: The issue of whether there might be an increased risk of cervical cancer associated with the use of oral contraceptives has been debated for decades. Early studies found a modest association with long-term use. A literature review was performed over the past 3 years, to establish whether there is any new evidence linking cervical cancer with the use of oral contraceptives. RECENT FINDINGS: A new analysis from eight studies conducted by the International Agency for Research on Cancer and a systematic review of cervical cancer and the use of hormonal contraceptives are two recent major epidemiological links strongly suggesting the increased risk of cervical cancer (up to twofold), but only for women who were both long-term users (5 years or more) and who had persistent human papilloma virus infections of the cervix. SUMMARY: These findings seem biologically plausible, but weighing the various risks and benefits, the World Health Organization does not recommend any change in oral contraceptive use or practice. PMID- 15128005 TI - Breast conservation in breast cancer: surgical and adjuvant considerations. AB - PURPOSE OF REVIEW: Breast-conserving surgery is accepted as an alternative to mastectomy for the treatment of early breast cancer. This paper reviews the recent contributions to the literature, with special emphasis on breast conserving surgery in the management of invasive breast cancer. RECENT FINDINGS: Long-term follow-up of randomized trials confirms that in women with early stage breast cancer, breast-conserving surgery achieves similar survival compared with mastectomy. Increase in early breast cancer detection, and efforts in patient and physician education are likely to enhance breast-conserving surgery use. Patients with locally advanced tumors may also become eligible for breast-conserving surgery after tumor downsizing with preoperative chemotherapy, with acceptable rates of ipsilateral breast tumor recurrence. Efforts should be made to minimize the risk of ipsilateral breast tumor recurrences after breast-conserving surgery as they have been associated with worse distant-disease-free and breast cancer specific survival rates. One of the most effective strategies to minimize ipsilateral breast tumor recurrence risk is to ensure negative surgical margins. This necessitates careful surgical planning, tumor localization, precise surgical technique and careful pathological processing. Radiation therapy is also a critical therapeutic tool to minimize the risk. Although the standard of care has been whole-breast irradiation, preliminary results with accelerated partial breast irradiation are promising. Adjuvant systemic therapy can further reduce the risk of ipsilateral breast tumor recurrence, but it cannot replace adequate surgery and radiation therapy for achieving local control after breast-conserving surgery. SUMMARY: Breast-conserving surgery is confirmed to be appropriate therapy for patients with early stage breast cancer. Increasing numbers of patients are likely to be eligible for surgery in the future. Strategies to minimize the risk of ipsilateral breast tumor recurrence while enhancing the convenience of breast-conserving surgery need to be pursued. PMID- 15128007 TI - Chemotherapy for early ovarian cancer. AB - PURPOSE OF REVIEW: The treatment of early ovarian cancer has long been based on non-randomized studies and on a small number of randomized studies without sufficient power. Adjuvant chemotherapy is often given to high-risk patients, but the benefit of such an approach has never been proven and the definition of high risk early ovarian cancer differs widely. Recently, the results of the two largest randomized clinical trials on early ovarian cancer became available. Both trials are discussed, and their results are related to the other relevant literature of the last three years. RECENT FINDINGS: A meta-analysis of over 1500 patients from the year 2001 confirmed tumor grade as a strong prognostic factor but it also demonstrated the adverse effect of capsule rupture before and during surgery. The Adjuvant Chemotherapy in Ovarian Neoplasm trial (European Organization for Research and Treatment of Cancer) randomized 448 patients to either adjuvant chemotherapy following surgery or observation. Adjuvant chemotherapy improved overall survival and disease-free survival in non-optimally staged patients but showed no benefit in optimally staged patients. The Medical Research Council International Collaborative Ovarian Neoplasm 1 trial randomized 477 patients in a similar way. Overall survival and disease-free survival were improved by adjuvant chemotherapy. It was argued that the study population of the International Collaborative Ovarian Neoplasm 1 trial probably represents non optimally staged patients, and this hypothesis explains why the results of this trial were in accord with those of the Adjuvant Chemotherapy in Ovarian Neoplasm trial. SUMMARY: The implications of these data are that a complete surgical staging is of utmost importance and should be pursued. In cases of non-optimal staging and contraindications for restaging, adjuvant chemotherapy is indicated to deal with unnoticed residual tumor deposits that exist in approximately 25% of cases. PMID- 15128006 TI - Chemotherapy for metastatic breast cancer. AB - PURPOSE OF REVIEW: This review is intended to summarize the most up-to-date information in the field of chemotherapy for advanced breast cancer. RECENT FINDINGS: During the past year, the literature on chemotherapy for metastatic breast cancer has focused on three main areas of interest: the development of new cytotoxic agents with most studies addressing the taxane-pretreated population; the evaluation of new combination regimens, mainly incorporating a taxane; and the development of new trastuzumab-cytotoxic-agent combinations in the HER-2 positive population. In addition, interesting data regarding combination versus sequential single-agent chemotherapy and the optimal duration of chemotherapy have been published. SUMMARY: Interesting new drugs have faced phase II development. Randomized trials will define their role in the daily management of metastatic breast cancer. New combination regimens, generally incorporating a taxane, improve short-term efficacy in comparison with standard anthracycline based combinations or single-agent chemotherapy, but at the cost of increased toxicity. Trastuzumab can be safely combined with several cytotoxic agents. PMID- 15128008 TI - Biomolecular prognostic factors in breast cancer. AB - PURPOSE OF REVIEW: To update clinicians on recent findings concerning the clinical usefulness of biomarkers in breast cancer, this review examines recently published papers dealing with promising prognostic/predictive biological factors. These factors can be classified according to their involvement in the main alterations characterizing tumor cells: self-sufficiency in growth signals, insensitivity to anti-growth signals, evasion of apoptosis, limitless replicative potential, sustained angiogenesis, and tissue invasion and metastasis. RECENT FINDINGS: Despite relevant research efforts and the identification of many putative good prognosticators, few of these factors are proving clinically useful for identifying patients at minimal risk of relapse, patients with a worse prognosis, or patients likely to benefit from specific treatments. Most of them, such as HER-2/neu, epidermal growth factor receptor, cyclin E, p53, bcl-2, vascular endothelial growth factor, urokinase-type plasminogen activator-1 and the recently discovered anti-apoptosis protein survivin, are suggested for possible inclusion in the category of biomarkers with a high level of clinico laboratory effectiveness. However, no single biomarker was able to identify those patients with the best (or worst) prognosis or those which would be responsive to a given therapy. Novel findings derived from gene-expression analysis indicate that the simultaneous consideration of molecular alterations contributing to the hallmarks of cancer might provide clinically useful prognostic, and perhaps therapeutic, information. SUMMARY: Rapid translation of laboratory findings to clinical practice was hampered by many difficulties, including technical and statistical concerns, a lack of assay standardization and comparability, and the modest design of translational studies. Many studies are performed on too small series of patients to provide reliable results; the studies are often heterogeneous in terms of treatment, patients and tumor characteristics, and data may be evaluated using different analytical approaches and are thus not easily comparable. Adequately planned prospective studies are required to assess the clinical utility of biomarker determinations. PMID- 15128009 TI - The use of sentinel lymph nodes in gynaecological malignancies. AB - PURPOSE OF REVIEW: Lymphatic mapping and sentinel node biopsy represent one of the most revolutionary advances in oncological surgery in recent years. In this review, the current state of sentinel node detection in gynaecological cancers and its use in vulvar and cervical cancer are assessed. RECENT FINDINGS: Since the recent clinical application of sentinel node biopsy for melanomas and breast cancer, there has been extensive research on the implementation of this technique to most solid neoplasias. Studies on the feasibility of sentinel node biopsy in vulvar cancer have shown that the status of the sentinel node is an accurate predictor of the status of inguinal nodes. The clinical implementation of the procedure requires validation and is under investigation. In the last two years, several pilot studies on the feasibility of lymphatic mapping/sentinel node biopsy in cervical cancer have yielded promising results. There is minimal experience of its use in endometrial cancer. Detailed pathological study of a sentinel node biopsy with ultrastaging and immunohistochemical or polymerase chain reaction analyses can identify lymph node micrometastasis that conventional methods would identify as negative for metastatic disease. The best histopathological procedure for sentinel node biopsy, the clinical significance of micrometastases, and the appropriate management of such micrometastases are currently under investigation. SUMMARY: Sentinel node biopsy is one of the main research interests in gynaecological oncological surgery. At present there are not enough data to permit modification of current treatment protocols. Large and multi-institutional trials are required in order to define the implementation of sentinel node biopsy in clinical practice with the objective of achieving safer and more conservative surgery. PMID- 15128010 TI - Modern management of vulvar cancer. AB - PURPOSE OF REVIEW: The radical surgical approach in the treatment of vulvar cancer patients has led to a favourable prognosis for the majority of patients with early stage vulvar cancer. However, morbidity is impressive, leading to more individualized treatment. The authors have reviewed the most recent literature on the pros and cons of the modifications in treatment, including surgery and primary radiotherapy, for primary squamous cell carcinoma of the vulva and vulvar melanoma. RECENT FINDINGS: The sentinel lymph node procedure is a promising method of staging in patients with early stage squamous cell carcinoma of the vulva and possibly for patients with vulvar melanoma, but its safety still has to be proved. Less radical surgery has led to a higher local and regional recurrence rate. There may be a role for primary radiotherapy of the groin in a selected group of patients. SUMMARY: The authors have concluded that the individualization of treatment for vulvar cancer patients has led to a decrease in morbidity but an increase in recurrences. The increase in recurrences does not appear to compromise prognosis, probably because of the lack of power, based on the low incidence of vulvar cancer. The sentinel lymph node procedure and primary radiotherapy are promising methods to reduce the morbidity of treatment but their safety needs to be studied in randomized trials. PMID- 15128011 TI - Hormone replacement therapy and risk of malignancy. AB - PURPOSE OF REVIEW: The fact that today our concern is oriented towards the risks rather than the benefits of hormone replacement therapy could be the clearest message about our current position. The safety of hormone replacement therapy, an estrogen-progestin combination which has been sympathetic to and supportive of disturbing menopausal symptoms of women, is seriously challenged. RECENT FINDINGS: Four randomized trials have now reported on the results of hormone replacement therapy in major potentially fatal conditions, in more than 20,000 women studied for about 5 years. The main concern regarding the increased risk of malignancy in healthy postmenopausal women in western countries has been breast cancer. It is estimated to cause an extra case in about six per 1000 users aged 50-59 and 12 per 1000 aged 60-69. Over the same period the estimated risk of endometrial cancer rates are not increased, with a relative risk of 0.76 per 1000 users aged 50-59. Overall, however, the increased incidence of malignancies is greater than any reduction, one per 230 users aged 50-59 and one per 150 aged 60 69. Randomized trials examining other important but rarer malignancies, like ovarian, gall bladder and urinary bladder cancer, are either nonexistent or too small to reliably describe any effects of hormone replacement therapy. SUMMARY: Conclusively epidemiological evidence suggests that hormone replacement therapy is associated with a small but substantial increase in breast cancer risk and combined estrogen-progesterone regimens further increase this hazard. Additionally, the evidence from the recent double blind placebo controlled randomized trial on the slight increase in the incidence of adverse cardiovascular events, has turned our orientation away from hormone replacement therapy as a long term therapy in postmenopausal women. In this review, the effort is to approach comprehensively and globally the information on the risks of hormone replacement therapy on several cancer sites. PMID- 15128012 TI - Recent developments in ovarian cancer genetics. AB - PURPOSE OF REVIEW: This review attempts to provide an update on recent research on inherited susceptibility to ovarian cancer. It covers articles mainly published in 2002 and 2003, with an emphasis on genetic counseling issues. RECENT FINDINGS: The major areas on which recent reports have focused include: (1) an expanded understanding of the BRCA1 and BRCA2 mutation spectrum and the frequencies of deleterious alleles in various ethnic groups; (2) investigations on how information is best transmitted to high-risk family members via genetic counseling; (3) an analysis of patient management changes based on genotype results; (4) social issues surrounding predictive testing for breast/ovarian cancer genes, including health insurance and discrimination concerns; and (5) an investigation into gynecologists' knowledge of ovarian cancer genetics, and their ability to provide genetic counseling for ovarian cancer to their patients. Preliminary reports from scientific meetings that have not yet been published in peer-reviewed journals are also discussed. SUMMARY: Recent developments in ovarian cancer genetics expand many of the areas that have been studied previously. A major focus of recent research has dealt with genetic counseling for families affected by hereditary breast and ovarian cancer. PMID- 15128013 TI - Immunological treatment of ovarian cancer. AB - PURPOSE OF REVIEW: Development of immunological treatments for ovarian cancer has not been a conspicuous success story over the past few years. Only a handful of clinical trials have reported immunological responses, and correlation with clinical benefit has been elusive. Several recent studies presented in this review, however, point to a revival of optimism for the development of novel immunotherapeutic strategies. RECENT FINDINGS: The cloning and sequencing of CA125, coupled with novel structural and functional insights, undoubtedly represent important steps forward. The possibility that CA125 could play a role in evasion of immunity by ovarian tumors may represent a new challenge, but does not detract from its potential as a therapeutic target. Of the recent clinical trial reports, the most intriguing results were seen from immunotherapy with a conventional mouse monoclonal antibody specific for CA125, in which human anti mouse antibody responses correlated significantly with improved survival of patients with advanced stage ovarian cancer and clinical evidence of recurrent disease at the time of treatment. SUMMARY: There is little doubt that CA125 will undergo a renaissance as an important target antigen for development of novel immunological treatments, particularly with regard to cellular therapies. Identification of other novel ovarian tumor antigens will also accelerate research focused on stimulation of T-cell immunity. Current research trends suggest a paradigm shift in emphasis from vaccines designed to elicit antibody responses to strategies such as dendritic cell vaccination that are designed to induce broader immunity, including ovarian tumor antigen-specific helper T lymphocyte and cytotoxic T-lymphocyte responses. PMID- 15128014 TI - Bibliography. Current world literature. Gynecologic oncology and pathology. PMID- 15128015 TI - Observation survey and invasive trial of chronic dialysis patients. PMID- 15128016 TI - An overview of regular dialysis treatment in Japan (as of 31 December 2001). AB - Questionnaire forms for an annual survey conducted at the end of 2001 were sent out to 3520 institutions, and 3485 replies were received (response rate, 99.00%). According to the survey, the dialysis population of Japan at year end was 219 183 patients, up 6.3% (13 049) over the year before. This equals 1721.9 dialysis patients per million population. The gross mortality rate was 9.3% for the year extending from the end of 2000 to the end of 2001. The mean age of patients beginning dialysis was 64.2 years (+/- 13.7 SD). The mean age of the overall dialysis population in the study year was 61.6 years (+/- 13.1 SD), which was also a higher age than the year before. Among dialysis patients, the primary disease was diabetic nephropathy in 38.1% of patients, slightly down from 39.1% the previous year. Chronic glomerulonephritis was the primary disease in 32.4% of cases, a decrease from 34.7% the previous year. This survey included for the first time the items of the lowest blood pressure during hemodialysis session, vasopressor therapy before dialysis and vasopressor therapy during dialysis session. An analysis of the relationship between the type of vascular access used at the initiation of dialysis and the survival prognosis revealed a significantly higher risk of death in patients undergoing dialysis with synthetic arterio venous (AV) fistula, AV shunt, or catheter implantation into a central vein than in those receiving dialysis treatments with a native fistula. There was a significantly lower risk of death in the patient group in whom the vascular access was created at 3-6 months before initiation of dialysis than in those in whom such access was created at the time of initiation or within 3 months before the initiation of dialysis. An analysis of the risk factors affecting survival prognosis in maintenance hemodialysis patients showed that risk factors for death are post-dialysis systolic blood pressure over 180 mm Hg and lower than 120 mm Hg, blood pressure elevating progressively from the start to the end of dialysis, serum high density lipoprotein cholesterol concentration of less than 30 mg/dL, and a higher ultrafiltration rate. In comparisons of the death risk between the patient group with a history of intervention for ischemic heart disease and the patient group with a history of myocardial infarction or heart failure but without such intervention, among diabetes patients, those who underwent percutaneous transluminal coronary angioplasty had a significantly lower risk of death than those in whom no intervention was made. PMID- 15128018 TI - Comparative analysis between dextran sulfate adsorption and direct adsorption of lipoproteins in their capability to reduce erythrocyte adhesiveness/aggregation in the peripheral blood. AB - The purpose of this study was to compare the degree of erythrocyte adhesiveness/aggregation (EAA) reduction of two low-density lipoprotein (LDL) apheretic procedures, namely direct adsorption of lipoproteins (DALI) and dextran sulfate adsorption (DSA). A significant (P < 0.001) reduction of EAA was noted in six hypercholesterolemic patients who underwent a total of 40 apheretic sessions and no difference was noted in the degree of EAA reduction by the two techniques. Thus. being a real-time and point-of-care test, the erythrocyte adhesiveness/aggregation test can be applied in relevant situations of acute ischemia, where therapeutic LDL apheresis could improve the hemorheology of individuals with increased concentrations of cholesterol and inflammatory sensitive proteins. PMID- 15128017 TI - The efficacy and safety of the new heparin-induced extracorporeal low-density lipoprotein precipitation system (Plasmat Futura) in comparison with the currently used system (Plasmat Secura). AB - The aim of the present study was to examine whether the recently introduced heparin-mediated extra-corporeal low-density lipoprotein precipitation (HELP) apheresis system Plasmat Futura (since 2001) was comparable to Plasmat Secura system, used to date, in its efficiency to remove atherogenic components, its ease of handling and operating as well as clinical safety and patient compliance. Coronary heart disease (CHD) patients (N = 21) were first treated with Plasmat Secura system and 13 of them were then randomly switched over to the upgraded Plasmat Futura system. Eight patients remained on Secura system. All together, 40 Futura treatments and 40 Secura treatments were performed. Blood samples were collected immediately before and after each apheresis therapy. Our data showed no significant differences in the reduction of plasma low-density lipoprotein, lipoprotein (a) and fibrinogen by Plasmat Futura and Secura system (P > 0.05). However, the major advantages of Plasmat Futura system are the ready-to-use sterile dialysis solutions instead of reverse osmosis device in Plasmat Secura, which ensures flexibility and lower risk of cross infections. Long-term tolerance and safety parameters showed no significant difference (P > 0.05). On the basis of our studies. Plasmat Futura system is easy to use, shows no adverse events and is comparable to Plasmat Secura in its capacity to remove proatherogenic plasma factors. PMID- 15128019 TI - Detection of specific IgE antibodies to nafamostat mesilate as an indication of possible adverse effects of leukocytapheresis using nafamostat mesilate as anticoagulant. AB - The aim was to determine whether adverse effects of leukocytapheresis (LCAP) are related to nafamostat mesilate (NM) as an anticoagulant. Anti-NM IgE were detected in inflammatory bowel disease (IBD) patients who were administrated LCAP in our institute. Forty-nine patients (ulcerative colitis (UC)/Crohn's disease (CD): 30/19) were evaluated. Anti-NM IgE was measured by the ELISA method. Total IgE level and eosinophil count was tested concurrently. We retrospectively checked the presence of allergic symptoms and medications used concurrently with LCAP. Anti-NM IgE were present in six symptomatic patients (6/49; 12.2%) whose adverse effects were highly suspected to be from NM. However, 21 patients showed anti NM IgE-negative, in spite of the fact that their adverse effects were also highly suspected to be from NM. Through the detection of anti-NM IgE alone we could not estimate the relevance of NM as an anticoagulant to the adverse effects of LCAP. PMID- 15128020 TI - Long daily hemodialysis sessions correct systemic complications of oxalosis prior to combined liver-kidney transplantation: case report. AB - An 18-year-old woman diagnosed with piridoxine-resistent primary hyperoxaluria type 1 (PH-1) and progressive renal insufficiency complicated with acute renal failure of obstructive origin who developed systemic oxalosis affecting the heart (cardiomyopathy), the skin (cutaneous ulcers) and vascular system (lower limb ischemia, as well as pulmonary and cerebral microcirculatory blockage resulting in pulmonary hemorrhage and tonic-clonic general seizures. As conventional hemodialysis (HD) or peritoneal dialysis (PD) are unable to eliminate enough oxalate to avoid a continuous positive balance, long daily sessions (6-7 h) of high-flux hemodialysis (highly permeable polyamide membrane of 2.1 m2) for 67 consecutive days normalized blood oxalate levels and reversed the systemic complications secondary to the calcium oxalate crystals deposit. The patient underwent a combined liver-kidney transplantation and has progressed well to the present time. The most important factors in PH-1 treatment are analyzed. Even though combined liver-kidney transplantation is the treatment of choice and should be performed before the glomerular filtrate rate (GFR) falls below 25 mL/min/1.73 m2, intensive HD becomes necessary to prevent oxalosis in the face of acute renal failure. Also, as our case shows, intensive HD can achieve a negative oxalate balance and reverse both the systemic lesions and the oxalate deposits. PMID- 15128021 TI - Review of combination of peritoneal dialysis and hemodialysis as a modality of treatment for end-stage renal disease. AB - Because the contribution of residual renal function (RRF) to total solute clearance is often significant in continuous ambulatory peritoneal dialysis (CAPD), loss of RRF over time can lead to inadequate dialysis if appropriate prescription management strategies are not pursued. Additionally, declines in ultrafiltration caused by increases in peritoneal permeability may limit continuation of CAPD therapy. Peritoneal dialysis and hemodialysis (PD + HD) combination therapy (complementary dialysis therapy) is an alternative method. This therapy allows the patient to maintain daily activities, as with CAPD, while undergoing once-a-week HD supplements for the insufficient removal of solutes and water. This therapy allows for the continuation of PD without shifting to total HD in PD patients who continue to have uremic symptoms even after individualization of the PD prescription. This treatment option is psychologically more acceptable to patients and may be expected to provide such accompanying beneficial effects as peritoneal resting, improvement of QOL and reduction in medical cost. PMID- 15128022 TI - Involvement of NADPH oxidase in hydrogen peroxide accumulation by Aspergillus niger elicitor-induced Taxus chinensis cell cultures. AB - After determining that hydrogen peroxide (H2O2) accumulation induced by a fungal elicitor from Aspergillus niger was from the superoxide dismutase-catalyzed dismutation of superoxide radical, the site of H2O2 generation in cell suspension cultures of Taxus chinensis was studied. The results showed that 90% and 10% of the elicitor-induced H2O2 accumulation respectively appeared in intracellular and extracellular fractions of cells, and that the elicitor-induced H2O2 accumulation in protoplasts and plasma membranes was similar to that in intact cells, indicating that the site of H2O2 accumulation was plasma membranes but not in extracellular fraction of Taxus cells. The H2O2 forming enzyme was also investigated. The elicitor-induced H2O2 accumulation in intact cells was not changed by loss of apoplastic peroxidase (POD) by the washing, and the H2O2 accumulation in plasma membranes was inhibited by the mammalian neutrophil NAD(P)H oxidase inhibitor diphenylene iodonium (DPI), but was slightly affected by exogenous POD and its inhibitor. Furthermore, in plasma membranes, the H2O2 accumulation was more significantly enhanced by NADPH than by NADH, and the former was more obviously decreased by DPI than the latter. The present results show that NADPH oxidase in plasma membranes is involved in H2O2 accumulation in fungal elicitor-induced Taxus chinensis cell cultures. PMID- 15128023 TI - Biosynthesis, accumulation and degradation of theobromine in developing Theobroma cacao fruits. AB - We have studied the purine alkaloid content and purine metabolism in Theobroma cacao fruits at differing growth stages: Stage A (young small fruit, fresh weight, ca. 2 g); stage B (medium size fruit, fresh weight, ca. 100 g) and stage C (large size, fresh weight, ca. 500 g). The major purine alkaloid in stage A fruits (mainly pericarp) was theobromine (0.7 micromol g(-1) fresh weight), followed by caffeine (0.09 micromol g(-1) fresh weight). The theobromine content of the pericarp decreased sharply with tissue age, and the caffeine content decreased gradually. A large amount of theobromine (22 micromol g(-1) fresh weight) had accumulated in seeds (mainly cotyledons) of stage C fruits. Theobromine was found also in the seed coat and placenta. Tracer experiments with [8-(14)C]adenine show that the major sites of theobromine synthesis are the young pericarp and cotyledons of T. cacao fruits. Limited amounts of purine alkaloids may be transported from the pericarp to seed tissue, but most purine alkaloids that accumulated in seeds appeared to be synthesised in cotyledons. Degradation of [8-(14)C]theobromine and [8-(14)C]caffeine to CO2 via 3-methylxanthine and ureides (allantoin and allantoic acid) was detected only in the pericarp of stage C fruits. PMID- 15128024 TI - Pectins from the albedo of immature lemon fruitlets have high water binding capacity. AB - The white part of citrus peel, the albedo, has a special role in water relations of both fruit and leaves from early on in fruit development. In times of drought, this tissue acts as a water reservoir for juice sacs, seeds and leaves. When water was injected into the albedo, free water was undetectable using magnetic resonance imaging. Microscopy showed tightly packed cells with little intercellular space, and thick cell walls. Cell wall material comprised 21% of the fresh albedo weight, and contained 26.1% galacturonic acid, the main constituent of pectin. From this, we postulated that pectin of the cell wall was responsible for the high water-binding capacity of the immature lemon albedo. Cell wall material was extracted using mild procedures that keep polymers intact, and four pectic fractions were recovered. Of these fractions, the SDS and chelator-soluble fractions showed viscosities ten and twenty times higher than laboratory-grade citrus pectin or the other albedo-derived pectins. The yield of these two pectins represented 28% of the cell walls and 62% of the galacturonic acid content of immature lemon albedo. We concluded that, from viscosity and abundance, these types of pectin account for the high water-binding capacity of this tissue. Compositional analyses showed that the two highly viscous pectic fractions differ in galacturonic acid content, degree of branching and length of side chains from the less viscous albedo-derived pectins. The most striking feature of these highly viscous pectins, however, was their high molecular weight distribution compared to the other pectic fractions. PMID- 15128025 TI - Ethylene-induced gene expression, enzyme activities, and water soaking in immature and ripe watermelon (Citrullus lanatus) fruit. AB - Watermelon fruit exhibit acute softening and placental-tissue water soaking following short exposure to exogenous ethylene. Experiments were performed to address transcript abundance and activities of cell wall and membrane hydrolases in placental tissue in response to treatment of watermelon fruit with ethylene. Watermelon fruit were harvested at immature and full-ripe stages and exposed to 50 microL L(-1) ethylene for 6 days at 20 degrees C. Ethylene affected the abundance of transcripts for PME (EC 3.2.1.11), and alpha-(EC 3.2.1.22) and beta GAL (EC 3.2.1.23) but these effects were dependent on fruit maturity and appeared not to be associated with the water-soaking syndrome. PG (EC 3.2.1.15) and EXP mRNAs accumulated significantly in response to ethylene exposure. Additionally, the levels of mRNA and activities of LOX (EC 1.13.11.12), PLC (EC 3.1.4.3) and PLD (EC 3.1.4.4) were elevated in fruit of both maturity classes exposed to ethylene and were temporally associated with the visible symptoms of water soaking. The activity trends and transcript abundance in ethylene- compared with air-treated fruit indicate that PG, EXP, LOX, PLC and PLD levels increase with the onset and development of the water-soaking disorder and support the view that catabolic reactions targeting the membranes and cell-walls contribute to the disorder. PMID- 15128026 TI - Abscisic acid and jasmonic acid affect proteinase inhibitor activities in barley leaves. AB - Proteinase inhibitor (PI) accumulation has been described as a plant defense response against insects and pathogens. The induction of PIs is known to be regulated by endogenous chemical factors including phytohormones. We studied the induction of barley chymotrypsin and trypsin inhibitory activities by aphid infestation, mechanical wounding, abscisic acid (ABA) and jasmonic acid (JA). Wounding experiments led to a minimal accumulation of PI activity (16% over controls) compared to that found in barley seedlings infested by aphids, where chymotrypsin inhibitor activity showed a two-fold increment. No systemic induction could be detected in healthy leaves of an infested or mechanically injured plant. Exogenous ABA applied on barley leaves increased the chymotrypsin inhibitory activity, while JA only increased trypsin inhibitory activity locally and systemically when applied exogenously. Our data suggest that two different mechanisms may be regulating the induction of these two types of inhibitors. PMID- 15128027 TI - Osmotic induced stimulation of the reduction of the viability dye 2,3,5 triphenyltetrazolium chloride by maize roots and callus cultures. AB - Live cells can reduce colorless 2,3,5-triphenyltetrazolium chloride (TTC) to a red insoluble compound, formazan. Maize (Zea mays) callus, when osmotically stressed by 0.53 mol/L mannitol, produced 7-times or more formazan than untreated control callus. This result was seen with all osmotica tested and could not be attributed to differences in TTC uptake rate or accumulation, increased respiration rate as measured by O2 uptake, or to de novo protein synthesis. Increased formazan production could be detected after 2.5 h of exposure to osmotic stress and leveled off after 48 h of exposure. The increased formazan production was only detected when callus was moved from high osmotic medium to low osmotic, TTC-containing medium. Abscisic acid increased TTC reduction only when added in combination with 0.53 mol/L mannitol. Incubation of maize seedling roots with 0.53 mol/L mannitol also increased formazan production as seen visually. Further studies are needed to determine the cause of the increased formazan production. These results show that TTC viability measurements must be carefully evaluated with appropriate controls to confirm their validity. PMID- 15128028 TI - Thermotolerance and antioxidant systems in Agrostis stolonifera: involvement of salicylic acid, abscisic acid, calcium, hydrogen peroxide, and ethylene. AB - This study investigated whether pre-treating plants with specific putative signaling components and heat acclimation would induce tolerance of a cool-season grass, creeping bentgrass (Agrostis stolonifera var. palustris), to subsequent heat stress and whether thermotolerance induction of those pretreatments was associated with the regulation of antioxidant regenerating enzymes. The treatments included foliar application of salicylic acid (SA), abscisic acid (ABA), calcium chloride (CaCl2), hydrogen peroxide (H2O2), 1-aminocyclopropane-1 carboxylic acid (ACC, a precursor of ethylene prior to the exposure of plants to heat stress (35 degrees C) in a growth chamber. Physiological measurements including turf quality, leaf photosynthetic rate, and levels of oxidative damage demonstrated that all treatments increased heat tolerance. The better heat tolerance for pre-treated plants as compared to controls was related to the protection of oxidative damage under heat stress. APX activity increased over the first 2 days and 5 days of heating for ACC and CaCl2 respectively, but for only 12 h for H2O2. SA and ABA pre-treatments had no effects on APX activity earlier, but maintained APX activity at a significantly higher level than in controls after 24 h of heating. SA and ABA pre-treatments had no effects on POX activity. ACC treatment significantly increased POX activity. Pre-treatment with CaCl2, H2O2, and HA reduced POX activity, particularly during the later phase of heating. Plants treated with SA, CaCl2, H2O2 and HA had lower CAT activity than their control plants prior to heating and within 48 h of heat stress. ABA and ACC pre-treatments maintained higher CAT activity than the controls after 48 h of heating. ACC, CaCl2, or HA pre-treatments increased SOD activity only before 5 days of heat stress. SA and ABA pre-treatments had less effect on APX activity earlier under heat stress. These results suggest that specific groups of potential signaling molecules may induce tolerance of creeping bentgrass to heat stress by reducing oxidative damage. PMID- 15128029 TI - Re-aeration-induced oxidative stress and antioxidative defenses in hypoxically pretreated lupine roots. AB - The level of free radicals and activities of antioxidative enzymes were examined in roots of lupine seedlings (Lupinus luteus L.) that were deprived of oxygen by subjecting them to root hypoxia for 48 and 72 h and then re-aerated for up to 24 h. Using electron paramagnetic resonance (EPR), we found that the exposure of previously hypoxically grown roots to air caused the increase in free radicals level, irrespective of duration of hypoxic pretreatment. Immediately after re aeration the level of free radicals was two times higher than in aerated control. The EPR signal with the g-values at the maximum absorption of 2.0057 and 2.0040 implied that the paramagnetic radicals are derived from a quinone. Directly after re-aeration of hypoxically pretreated roots, the activity of superoxide dismutase (SOD, EC 1.15.1.1) increased to its highest value, followed by a decline below the initial level, whereas activities of catalase (CAT, EC 1.11.1.6) and peroxidase (POX, EC 1.11.1.7) were diminished or only slightly influenced during re-aeration. The electrophoretic patterns of the soluble extracts show 4 isozymes of SOD, 4 isozymes of POX and 1 isozyme of CAT. The level of H2O2 was enhanced or lowered by re-aeration, depending on the previous duration of hypoxia. At the onset of re-aeration products of lipid peroxidation were present at a three fourth of the levels found in aerobic control. Their levels increased after prolonged exposure to air but remained lower than those in aerobic control even after 24 h of re-aeration. Re-admission of oxygen resulted in about 20% rise in oxygen uptake by root axes segments immediately after transfer of roots from hypoxia and the high uptake rates were observed over whole re-aeration period. Oxygen consumption by root tips was significantly reduced just after transfer from hypoxic conditions as compared to aerated control but after 24 h of re aeration even approached the control level. The results are discussed in relation to the ability of lupine roots to cope with oxidative stress caused by re aeration following hypoxic pretreatment. PMID- 15128030 TI - The rapid yellowing of spruce at a mountain site in the Central Black Forest (Germany). Combined effects of Mg deficiency and ozone on biochemical, physiological and structural properties of the chloroplasts. AB - Biochemical, physiological and ultrastructural changes of the chloroplasts were examined in the course of the rapid yellowing process of spruce (Picea abies (L.) Karst.) at a Mg-deficient and ozone polluted mountain site (Schollkopf mountain, Central Black Forest, Germany, 840 m a.s.l.). While at an early stage of yellowing the chlorophyll (Chl) content of the needles decreased slowly, significant changes occurred in the chloroplasts: The lability of the light harvesting Chl a/b protein complex LHC II increased; the thylakoid cross sectional area of chloroplasts in the outer mesophyll of the needles decreased, and their Chl fluorescence showed typical changes like the decrease of Fv/Fm and the increase of the photoinhibitory Fv quenching. Later on, the Chl content decreased rapidly, the changes in the chloroplasts continued and the needles turned yellow. Lutein and the pigments of the xanthophyll cycle were enhanced in relation to Chl a. Light and dark reactions of the xanthophyll cycle were highly active indicating efficient proton pumping and NADPH formation. The ratio of nonappressed to appressed thylakoid membranes increased with decreasing Fv/Fm suggesting that structural and fluorescence properties of the chloroplasts were related. The response of the needles to defined shading and improved Mg supply was also examined. The combined effects of strong sun light, low levels of non Chl-bound Mg (Mg(free)) and ozone concentrations exceeding 80 microg m(-3) are shown to be necessary to induce the rapid yellowing process. For needles with Mg(free) < 0.12 mg g(-1) needle dry matter, the lability of the LHC II was correlated with the ozone concentration suggesting that the destabilization of the LHC II plays a central role in the rapid yellowing process. PMID- 15128031 TI - MADS-box genes from perennial ryegrass differentially expressed during transition from vegetative to reproductive growth. AB - In contrast to well-studied dicot plants like Arabidopsis and Antirrhinum, relatively few genes controlling the transition to flowering and flower development of agronomically important monocot species have been identified. In perennial ryegrass (Lolium perenne) the transition from vegetative to reproductive growth is triggered by an obligate vernalization period (primary induction) of at least 12 weeks at temperatures below 5 degrees C under short days, followed by increased temperature and day length (secondary induction). Here we report the isolation of nine ryegrass MADS-box (LpMADS) genes by a differential display method specific to this family of transcription factors. Three of the nine MADS-box genes show homology to the APETALA 1 (AP1) subfamily, two to the SEPALLATA (SEP) subfamily, one to the AGAMOUS-LIKE 6 (AGL6) subfamily, and three show homology to the newly identified OsMADS1 subfamily. The three AP1 homologues are up-regulated, both in the shoot apex and in leaves, in response to vernalization, while expression of the other six are increased by secondary induction during inflorescence development, although not in leaves. Differences in the sequence and hierarchy of flowering gene expression patterns indicate that the Arabidopsis-based flowering model is not completely applicable to explain the molecular events leading to the floral transition in grasses. PMID- 15128032 TI - A lipid transfer protein gene BG-14 is differentially regulated by abiotic stress, ABA, anisomycin, and sphingosine in bromegrass (Bromus inermis). AB - The objective was to investigate the expression of a lipid transfer protein gene (LTP) both in bromegrass (Bromus inermis) cells and seedlings after exposure to abiotic stresses, abscisic acid (ABA), anisomycin, and sphingosine. A full-length cDNA clone BG-14 isolated from bromegrass suspension cell culture encodes a polypeptide of 124 amino acids with typical LTP characteristics, such as a conserved arrangement of cysteine residues. During active stages of cold acclimation LTP expression was up-regulated, whereas at the final stage of cold acclimation LTP transcript level declined to pre-acclimation level. A severe drought stress induced the LTP gene; yet, LTP expression doubled 3 d after re hydration. Both temperature and heat shock duration influence LTP induction; however temperature is the primary factor. Treatment with NaCl stimulated accumulation of LTP mRNA within 15 min and the transcripts remained at elevated levels for the duration of the salinity stress. Most interestingly, Northern blots showed LTP was rapidly induced not only by ABA, but also by anisomycin and sphingosine in suspension cell cultures. Of the three chemicals, ABA induced the most rapid and highest response in LTP expression as well as highest freezing tolerance, whereas sphingosine was the least active for both LTP expression and freezing tolerance. PMID- 15128033 TI - Salicylic acid-, but not cytokinin-induced, resistance to WClMV is associated with increased expression of SA-dependent resistance genes in Phaseolus vulgaris. AB - Two-week-old Phaseolus vulgaris plants, wick-fed with 1 mmol/L salicylic acid (SA) or 50 nmol/L dihydrozeatin (DHZ), showed partial inhibition of the accumulation of white clover mosaic virus (WClMV) in infected primary leaves. This inhibition was measured as a decrease in the accumulation of both viral mRNA and viral coat protein, especially at the early stages of infection. Salicylic acid treatment resulted in moderately increased expression of phenylalanine ammonia lyase (PAL), NPR1, PR1 and HSP70 genes that participate in resistance to pathogens in plants. In contrast, DHZ treatments did not induce significant changes in expression of these genes. The expression of the P. vulgaris alternative oxidase (AOX) gene homolog, an enzyme implicated in plant resistance to viruses, showed low constitutive expression during the first 11 days post infection and was not affected by either SA or DHZ. It appears that, while SA induced the NPR1-PR1 pathogen defense pathway genes, both SA and DHZ may use a different pathway to induce resistance to WClMV infection in P. vulgaris plants. PMID- 15128034 TI - Transcriptome changes in foxtail millet genotypes at high salinity: identification and characterization of a PHGPX gene specifically upregulated by NaCl in a salt-tolerant line. AB - Using a macro array filter with 711 cDNA inserts representing 620 unigenes selected from a barley EST collection, we identified transcripts differentially expressed in salt (NaCl)-treated tolerant (cv. Prasad) and sensitive (cv. Lepakshi) seedlings of foxtail millet (Setaria italica L.). Transcripts of hydrogen peroxide scavenging enzymes such as phospholipid hydroperoxide glutathione peroxidase (PHGPX), ascorbate peroxidase (APX) and catalase 1 (CAT1) in addition to some genes of cellular metabolism were found to be especially up regulated at high salinity in the tolerant line. To analyse this process at the protein level we examined protein expression patterns under various stress conditions. A 25 kD protein with a pI of 4.8 was found to be induced prominently under high salt concentrations (250 mmol/L). This salt-induced 25 kD protein has been purified and identified by peptide sequencing as PHGPX protein. The increase of the PHGPX protein level under salt stress in the tolerant line parallels the PHGPX mRNA results of array analysis but was more pronounced. We cloned and characterized the foxtail millet PHGPX cDNA, which shows 85% and 95% homology at the DNA and protein level, respectively, to one stress-induced member of the small barley PHGPX gene family encoding non-selenium glutathione peroxidases. As shown by Southern blot analysis, a small family of PHGPX genes exists in foxtail millet, too. The specific expression pattern of the PHGPX gene in salt-induced tolerant millet seedlings suggests that its product plays an important role in the defense reaction against salt-induced oxidative damage and that the characterized glutathione peroxidase is one of the components conferring resistance against salt to the tolerant foxtail millet cultivar. PMID- 15128035 TI - Genetic and epigenetic evaluations of citrus calluses recovered from slow-growth culture. AB - The embryogenic callus of "Red Marsh" grapefruit was stored in vitro by slow growth culture method for one year, and survived with a significant weight increment over that period. The survivers regenerated somatic embryos more easily than the controls. Eight callus lines were used for genetic analyses. Although chromosome number variations were verified by cytological examination both in the controls and the stored samples, the ploidy level remained relatively stable during the storage period. Randomly amplified polymorphic DNA (RAPD) analysis was performed to detect DNA sequence variation. No difference in RAPD pattern was found with the 102 primers used. However, a methylation sensitive amplified polymorphism (MSAP) assay showed DNA methylation changes in the stored samples compared with the controls. PMID- 15128036 TI - Acetyl coenzyme A concentrations in plant tissues. AB - Despite the importance of acetyl coenzyme A in many facets of metabolism and the availability of methods for estimation of its concentration, data for acetyl-CoA concentrations in plant tissues have been very scarce. A method using reversed phase HPLC for the quantitative estimation of acetyl-CoA was applied to a variety of plant tissues. In three different developing oilseeds the bulk acetyl-CoA concentration ranged from 5 to 25 nmoles/g fresh weight. In Arabidopsis thaliana leaves it was 5 nmoles/g fresh weight, and in Spinacia oleracea leaves 6.8 nmoles/g fresh weight. Immediate quenching of the harvested tissue in liquid nitrogen is needed to obtain high recoveries of acetyl-CoA. PMID- 15128037 TI - Triacontanol negatively modulates the jasmonic acid-stimulated proteinase inhibitors in tomato (Lycopersicon esculentum). AB - Triacontanol (TRIA), a long chain aliphatic alcohol (C30H61OH) reverses the effect of jasmonic acid (JA) in inducing proteinase inhibitors (PIs) in tomato leaves. Porcine pancreas trypsin and Spodoptera litura gut proteinases were inhibited in the presence of leaf proteins treated with JA, and TRIA partially reverses this effect. Spodoptera litura larvae fed with tomato leaves treated with JA were reduced in body weight and TRIA is able to partially reverse this JA induced effect. These results reflect the partial reversal effect of TRIA in down regulating the JA-induced production of proteinase inhibitors. PMID- 15128038 TI - Root enzyme activities associated with resistance to Heterodera avenae conferred by gene Cre7 in a wheat/Aegilops triuncialis introgression line. AB - The effect of Cereal cyst nematode (Heterodera avenae) infection on the expression of putative root defence-related enzymes, peroxidase (PER), esterase (EST) and superoxide dismutase (SOD), was studied in roots of a wheat/Aegilops triuncialis introgression line TR-3531 carrying the Cre7 resistance gene. We analysed detoxificant isozyme changes within roots of the resistant line and their susceptible parent (H-10-15) as a control, during the early interaction with the pathotype Ha71 of H. avenae. Isoelectrofocusing (IEF) isozyme analysis, four and seven days after infection, revealed that PER, EST and SOD activities increased in the resistant line TR-3531 in comparison with the susceptible control. Moreover, four and seven days after infection, the TR-3531 line showed the expression of new PER isozymes, with pIs of 9.7, 9.0, 8.5, 6.5 and 5.0, and an increased activity of some constitutive isoforms. The intensity of some EST and SOD constitutive bands increased in the resistant line after infection. However, no new isoforms were detected for EST and SOD systems. Nematode-induced enzyme activity was minor (PER) or did not occur (EST and SOD) in the compatible interaction with H-10-15. The enhanced peroxidase and esterase activities may play a role in the lignification of cell walls, which assists in the resistance to penetration by the nematode. PMID- 15128039 TI - Thermodynamic, conformational and functional properties of the human C1q globular heads in the intact C1q molecule in solution. AB - Thermodynamic. conformational and functional properties of the human C1q globular heads (hgC1q) were studied with the experimental approaches, which allow investigating these properties in the intact hC1q molecule in solution. Surprisingly, the scanning calorimetry data reveal a low level of cooperativity of interactions between the hgC1q A, B and C domains even at a neutral pH area. Ionization of His residues due to acidification of the medium at the pH range from 6 to 5 or the chemical modification of His residues completely abolishes the cooperative interactions between the domains without significant effect on their conformation. The thermodynamic data provide evidence that the hgC1q module is composed of three structurally independent A, B and C globular domains characterized by the practically identical thermal stability and very similar enthalpy of melting. The spectroscopic studies and modification with 2-oxy-5 nitrobenzylbromide (ONBB) indicate that Trp residues in the hgC1q A and C domains are accessible to the solvent that has been confirmed by the hgC1q crystal structure solved and refined to 1.9 A. The modification of Trp residues significantly affects the complement-dependent cytotoxicity without noticeable effect on the hC1q conformation. These data provide evidence that Trp residues are the components of immunoglobulin-binding sites both in the hgC1q A and C domains. PMID- 15128040 TI - Functional, molecular and structural characterisation of five anti-Brucella LPS mAb. AB - The O-antigen of the gram negative bacteria Brucella is composed of an homopolymer of 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl (or perosamine). Several mAb interact specifically with only the O-antigen of certain Brucella species. Although, many studies show that this specific recognition results mainly from the ratios of alpha 1-2 and alpha 1-3 link between the different Brucella strain perosamine residues, little is known about the mAb recognising this O-antigen. In this paper, we describe the binding profile of five anti Brucella O-antigen mAb to the LPS of two Brucella strains and a bacteria possessing a nearly identical O-antigen: Yersinia enterocolitica 0:9. We show that the specificity of these five mAb can be correlated to their germ line gene usage. Besides, their relative affinity to the different LPS is correlated to their ability to protect against Brucella infection by passive transfer in a mouse model. The analysis of their 3D structure gives new hypothesis of the epitopes recognised. PMID- 15128041 TI - Molecular proximity of complement receptor type 3 (CR3) and the glycosylphosphatidylinositol-linked protein GPI-80 on neutrophils: effects of cell adherence, exogenous saccharides, and lipid raft disrupting agents. AB - GPI-80, a novel glycosylphosphatidylinositol (GPI)-anchored protein on polymorphonuclear leukocytes, has been reported to cooperate with CR3 in several aspects of cell function including cell activation, adhesion and migration. The present study investigates the physical proximity of CR3 and GPI-80 on living cells using resonance energy transfer (RET) techniques, which gives positive results when the separation distance is < or = 7 nm. RET from donor-labeled anti CR3 to acceptor-labeled anti-GPI-80 was detected on adherent neutrophils, but not observed for non-adherent cells. Furthermore, RET was not observed on cells treated with cell adhesion inhibitors 4-bromophenacyl bromide (BPB), N ethylmaleimide (NEM) or cytochalasin D, suggesting dynamic interactions between CR3 and GPI-80. CR3-to-GPI-80 proximity was blocked by N-acetyl-D-glucosamine (NADG), but not by other monosaccharides such as D-mannose, fructose, fucose, glucose, sorbitol, or galactose; molecular proximity was also disrupted by the glycolipid raft depleting agents 2-OH-propyl-betaCD and MbetaCD. Thus, lipid rafts may be important for the physical and functional cooperation of CR3 and GPI 80. PMID- 15128042 TI - The Filamin-A is a partner of Tc-mip, a new adapter protein involved in c-maf dependent Th2 signaling pathway. AB - Using a yeast two-hybrid screen, we identified Filamin-A as a binding partner of the new adapter protein c-mip (c-maf inducing protein) and it's splice variant Tc mip (truncated c-maf inducing protein). We have previously shown that Tc-mip is involved in Th2 signaling pathway and cytoskeletal reorganization in patients with minimal change nephrotic syndrome (MCNS), the most frequent glomerular disease in children. We showed that Filamin-A and c-mip or Tc-mip co immunoprecipitate from c-mip or Tc-mip Jurkat transfected cells using antibodies directed against both types of proteins. In co-immunoprecipitate Jurkat cells, Filamin-A and c-mip were distributed evenly in the cytoplasm, whereas in Tc-mip transfected Jurkat cells, Filamin-A was expressed in zones facing the cell contact. Moreover, we found that Filamin-A was upregulated in T lymphocytes of MCNS patients, as compared to normal subjects. These findings suggest that Filamin-A interacts with c-mip/Tc-mip in this new T-cell signaling pathway. PMID- 15128043 TI - Three-dimensional modeling of a pre-B-cell receptor. AB - Signals delivered by the immunoglobulin (Ig)-like pre-B-cell receptor (pre-BCR) are critical for efficient maturation of early precursor B (pre-B) cells. A pre BCR contains two immunoglobulin mu-heavy chains (muHC), two surrogate light chains (SLC) consisting of the non-covalently associated polypeptides, VpreB and lambda5, and the heterodimeric signaling transducer Igalpha/beta. Although, it is generally accepted that signals initiated from the pre-BCR are required for efficient expansion and differentiation of pre-B cells, the three-dimensional structure of this receptor has not yet been determined by either NMR or X-ray spectroscopy. Therefore, we used indirect computer-assisted molecular modeling techniques to predict for the first time three-dimensional coordinates of the pre BCR, the conformation of the SLC components, VpreB and lambda5, and the position and flexibility of the so-called non-Ig-like unique tails at the C-terminus of VpreB and the N-terminus of lambda5. Structure prediction revealed that these unique tails of VpreB and lambda5 protrude from the SLC at the position where the CDR3 of a conventional IgLchain would be located. Thus, the unique tails are accessible for ligand binding, which supports the recent finding that the lambda5 unique tail is required for pre-BCR/stroma cell interaction. Further, the non covalent interaction of the extra beta-strand of lambda5 (beta8) with VpreB is predicted to result in a stabilization of the tertiary structure of VpreB. In summary, three-dimensional computer modeling suggests that the structure of a pre BCR resembles that of a conventional B-cell receptor (BCR) and that the lambda5 unique tail could be a major binding site for pre-BCR ligands. PMID- 15128044 TI - Analysis of strand biased 'G'.C hypermutation in human immunoglobulin V(lambda) gene segments suggests that both DNA strands are targets for deamination by activation-induced cytidine deaminase. AB - Somatic hypermutation. which diversifies the immunoglobulin repertoire by introducing mutations into rearranged IgV genes, is dependent on the expression of activation-induced cytidine deaminase (AID). It has been proposed that AID deaminates DNA directly, generating mutations at C bases. Mutations from C and G are linked, and it has been suggested that mutations from G arise either during repair of DNA following deamination of C or by deamination of C on both DNA strands. Studies demonstrating that AID deaminates ssDNA on the non-transcribed strand support the former hypothesis. However, analyses of microsequences surrounding mutations suggest that the G.C mutator acts on both DNA strands equivalently. Unusually, in human IgV(lambda) genes, there is G.C strand bias favoring mutation from G. In IgV(lambda), 92% of mutations from G occur in GNW motifs. Hotspots for mutation from G IgV(lambda) are often independent of C nucleotides in the context of local microsequence. This independence of G and C mutation, yet retained dependence on local microsequence suggests that mutations from G arise independent of C on the non-transcribed strand. We suggest that both DNA strands are deaminated and that the transcribed strand is preferentially deaminated in human IgV(lambda) resulting in bias towards mutations from G. PMID- 15128045 TI - Role of calreticulin from parasites in its interaction with vertebrate hosts. AB - Although parasites range from protozoan to complex, evolutionary advanced arthropods, in general, a hallmark of parasite life cycles is their ability to adapt to changes in temperature, pH and host defense strategies. Calreticulin, a calcium-binding protein, highly conserved and multifunctional, is present in every cell of higher organisms, except erythrocytes. The surprising array of calreticulin-associated functions include lectin-like chaperoning, calcium storage and signaling, modulation of gene expression, cell adhesion, enhancement of phagocytosis of C1q or collectin opsonized apoptotic cells, inhibition of angiogenesis and tumoral growth, inhibition of perforin pore formation in T and NK cells, and inhibition of C1q-dependent complement activation. Likewise, calreticulin is present in a wide spectrum of sub cellular compartments. Parasite calreticulin shows a surprisingly high degree of conservation within the framework of its functional domains. Its role within the parasite/host relationship needs to be assessed further, in particular with regard to its impact on parasite infectivity, by helping to evade from its hosts' immune response. With special emphasis on calreticulin from Trypanosoma cruzi, the intracellular protozoan agent of American trypanosomiasis (Chagas' disease), we wish to exemplify and highlight the various implications of parasite calreticulin, within the pathophysiology of parasite-mediated human and animal disease. PMID- 15128046 TI - Comparison of cytochrome P450 (CYP) genes from the mouse and human genomes, including nomenclature recommendations for genes, pseudogenes and alternative splice variants. AB - OBJECTIVES: Completion of both the mouse and human genome sequences in the private and public sectors has prompted comparison between the two species at multiple levels. This review summarizes the cytochrome P450 (CYP) gene superfamily. For the first time, we have the ability to compare complete sets of CYP genes from two mammals. Use of the mouse as a model mammal, and as a surrogate for human biology, assumes reasonable similarity between the two. It is therefore of interest to catalog the genetic similarities and differences, and to clarify the limits of extrapolation from mouse to human. METHODS: Data-mining methods have been used to find all the mouse and human CYP sequences; this includes 102 putatively functional genes and 88 pseudogenes in the mouse, and 57 putatively functional genes and 58 pseudogenes in the human. Comparison is made between all these genes, especially the seven main CYP gene clusters. RESULTS AND CONCLUSIONS: The seven CYP clusters are greatly expanded in the mouse with 72 functional genes versus only 27 in the human, while many pseudogenes are present; presumably this phenomenon will be seen in many other gene superfamily clusters. Complete identification of all pseudogene sequences is likely to be clinically important, because some of these highly similar exons can interfere with PCR based genotyping assays. A naming procedure for each of four categories of CYP pseudogenes is proposed, and we encourage various gene nomenclature committees to consider seriously the adoption and application of this pseudogene nomenclature system. PMID- 15128047 TI - Effects of CYP2C9 polymorphisms on the pharmacokinetics of R- and S-phenprocoumon in healthy volunteers. AB - CYP2C9 catalyses the biotransformation of the oral anticoagulants S-warfarin and R- and S-acenocoumarol. According to data obtained in vitro, phenprocoumon is also metabolized by CYP2C9 but the impact of the CYP2C9 polymorphism on phenprocoumon pharmacokinetics has not been studied. Twenty-six healthy heterozygous and homozygous carriers of the CYP2C9 alleles *1 (wild-type), *2 (Arg144Cys), and *3 (Ile359Leu) received a single oral dose of 12 mg of racemic phenprocoumon. Plasma and 12 h urine concentrations of both enantiomers and their monohydroxylated metabolites were measured by high-performance liquid chromatography with mass spectrometry detection. No significant effect of the CYP2C9 variants *2 and *3 on R-phenprocoumon pharmacokinetic parameters was detected, but S-phenprocoumon clearance tended to decrease with increasing number of CYP2C9*2 and *3 alleles. The ratios of S- to R-phenprocoumon plasma clearances were higher with a median of 0.95 in carriers of *1/*1 versus 0.65 in *3/*3 (P < 0.001 for trend). Plasma and urine concentrations of 4'-, 6- and 7 hydroxyphenprocoumon were significantly lower in homozygous carriers of the CYP2C9*2 and *3 variants compared to CYP2C9*1/*1. Carriers of CYP2C9*3/*3 had a median AUC of (R,S) 7-OH-phenprocoumon of only approximately 25% compared to the wild-type genotype. The AUC of (R,S) 6-OH-phenprocoumon was only approximately 50% in CYP2C9*3/*3 compared to the homozygous wild-type genotype. In conclusion, carriers of CYP2C9*2 and *3 alleles had a lower metabolic capacity regarding phenprocoumon hydroxylation than those with CYP2C9*1/*1. However, regarding phenprocoumon hydroxylation CYP2C9 genotypes had only marginal effects on S- and R-phenprocoumon total clearance in healthy volunteers. PMID- 15128048 TI - The risk of overanticoagulation in patients with cytochrome P450 CYP2C9*2 or CYP2C9*3 alleles on acenocoumarol or phenprocoumon. AB - Cytochrome P4502C9 (CYP2C9) is the main enzyme implicated in coumarin anticoagulant metabolism. The variant alleles CYP2C9*2 and CYP2C9*3 are associated with an increased response to warfarin. However, an effect on acenocoumarol dose requirements appears to be absent for the CYP2C9*2 allele and the consequences for the metabolism of phenprocoumon have not yet been established. We investigated CYP2C9 polymorphisms in relation to the international normalized ratio (INR) during the first 6 weeks of treatment and its effect on the maintenance dose in a cohort of 1124 patients from the Rotterdam Study who were treated with acenocoumarol or phenprocoumon. There was a statistically significant difference in first INR between patients with variant genotypes and those with the wild-type. Almost all acenocoumarol-treated patients with a variant genotype had a significantly higher mean INR and had a higher risk of an INR > or = 6.0 during the first 6 weeks of treatment. A clear genotype-dose relationship was found for acenocoumarol-treated patients. For patients on phenprocoumon, no significant differences were found between variant genotypes and the wild-type genotype. Individuals with one or more CYP2C9*2 or CYP2C9*3 allele(s) require a significantly lower dose of acenocoumarol compared to wild type patients. Phenprocoumon appears to be a clinically useful alternative in patients carrying the CYP2C9*2 and *3 alleles. PMID- 15128049 TI - Human glutathione S-transferase A2 polymorphisms: variant expression, distribution in prostate cancer cases/controls and a novel form. AB - Variability of expression of the major glutathione S-transferases (GSTs) of liver, GSTA1 and GSTA2, is thought to affect the efficiency of detoxification of xenobiotics, including chemical carcinogens. Polymorphism of the GSTA1 regulatory sequence determines some of the variation of hepatic GSTA1 expression, but the polymorphisms in GSTA2 (exons 5 and 7) were not thought to affect GSTA2 activity. By examining GST protein expression for a set of human liver and pancreas samples (coupled with a cloning/polymerase chain reaction-restriction fragment length polymorphism strategy), we identified a novel substitution Pro110Ser (328C>T) and the corresponding novel variant GSTA2*E (Ser110Ser112Lys196Glu210), and confirmed the presence of variants GSTA2*A (Pro110Ser112Lys196Glu210), GSTA2*B (Pro110Ser112Lys196Ala210) and GSTA2*C (Pro110Thr112Lys196Glu210). GSTA2*C occurred at 30-60% (i.e. approximately 100-fold more frequent than previously reported) and GSTA2*E occurred (heterozygous) at approximately 11%. Hepatic expression of the Ser112 variants (GSTA2*A, GSTA2*B or GSTA2*E) was approximately four-fold higher than that of the Thr112 variant (GSTA2*C). Compared to any other variant, GSTA2E had lower rates of catalysis towards 1-chloro-2,4-dinitrobenzene (CDNB), 4-vinylpyridine, and cumene-, t-butyl- and arachidonic acid hydroperoxides, although kcat/Km for CDNB were similar for all four variants. Using a prostate cancer case-control population, it was found that GSTA1*A/GSTA2 C335 and GSTA1*B/GSTA2 G335 were in linkage disequilibrium in Caucasians but not in African-Americans. However, there were no significant differences in the distribution of these polymorphisms or resultant haplotypes by case status. Nevertheless, the rare genotypes, GSTA2*E/*E and GSTA1*B/*B + GSTA2*C/*C (potential low GSTA2 activity and low hepatic GSTA1 and GSTA2 expression, respectively) could increase the risk of adverse effects of xenobiotics via compromised efficiency of detoxification. PMID- 15128050 TI - Promoter polymorphisms in glutathione-S-transferase genes affect transcription. AB - The glutathione-S-transferases are a group of enzymes that play a major role in detoxification and defence against toxic, carcinogenic and other compounds. We analysed the proximal promoters of 14 genes encoding glutathione-S-transferase for polymorphism. Ten of the promoters contained sequence variants, nine of which we were able to clone into a reporter gene vector, pGL3. The relative ability of each haplotype to promote transcription of the luciferase gene was tested in each of two human cell lines (HEK293t and TE671) using a cotransfected CMV-SEAP plasmid as a control. Four genes (GSTA1, GSTA2, GSTM4 and GSTT2) showed activity differences greater than 1.5-fold between haplotypes, and a fifth gene (MGST1) showed a 1.4-fold difference. The promoter sequence variants in these genes may therefore play a role in human variation, susceptibility to diseases and the effects of drugs. PMID- 15128051 TI - Pharmacoeconomic evaluation of testing for angiotensin-converting enzyme genotype before starting beta-hydroxy-beta-methylglutaryl coenzyme A reductase inhibitor therapy in men. AB - This study aimed to assess the potential cost-effectiveness of screening men for their angiotensin-converting enzyme (ACE)-genotype before starting statin therapy. We used a combination of decision-analytic and Markov modelling techniques to evaluate the long-term incremental clinical and economic effects associated with genetic testing of men with hypercholesterolemia before starting treatment with statins. The study was performed from a health care payer perspective. We used data from the Rotterdam study, a prospective population based cohort study in the Netherlands, which was started in 1990 and included 7983 subjects aged 55 years and older. Men treated with cholesterol-lowering drugs at baseline or with a baseline total cholesterol > or = 6.5 mmol/l were included. The ratio of difference in lifelong costs between the screening strategy and the no screening strategy to difference in life expectancy between these strategies was calculated. We also performed a cost-utility analysis. The base case was a 55-year-old man with hypercholesterolemia who was initially untreated. Several univariate sensitivity analyses were performed. All costs were discounted with an annual rate of 5%. Screening men for their ACE-genotype was the dominant strategy for the base case analysis, because the screening strategy saved money (851 Euro), but life expectancy was not changed. Screening was the dominant strategy for all age-groups in our cohort. Even in 80-year-old subjects, with the shortest life-expectancy, it was cheaper to screen than to give lifelong treatment to men with a DD genotype without success. Even if all DD subjects were treated with other (non-statin) cholesterol-lowering drugs, screening remained the cost-effective strategy. The results of the cost-utility analysis were similar. Discounting the effects with 5% per year also had no major impact on the conclusions. If other studies confirm that men with the DD genotype do not benefit from treatment with statins, screening for ACE genotype in men most likely will be a cost-effective strategy before initiating statin therapy. PMID- 15128052 TI - Effects of PPARalpha, gamma and delta haplotypes on plasma levels of lipids, severity and progression of coronary atherosclerosis and response to statin therapy in the lipoprotein coronary atherosclerosis study. AB - Peroxisome proliferator-activated receptors (PPARs) alpha, delta and gamma are nuclear transcription factors that regulate fatty acid biosynthesis. Our objectives were to determine the effects of PPAR haplotypes on biochemical, angiographic, clinical phenotypes and their responses to treatment with fluvastatin. We genotyped 372 Lipoprotein and Coronary Atherosclerosis Study subjects for seven single nucleotide polymorphisms (SNPs) in PPARalpha (-35 089A>C, 484C>G), delta (-4401C>T, 294T>C) and gamma (34C>G, 25 506C>T, 161C>T) by restriction mapping or 5' exonuclease assay. We reconstructed and estimated haplotypes frequencies using four algorithms. Linkage disequilibrium (LD) was calculated by D' and haplotype effects by permutation and regression analyses. The PPARD and PPARG SNPs were in LD. The baseline plasma triglyceride levels and their responses to treatment with fluvastatin were associated with PPARD haplotypes (P = 0.01). Triglyceride levels were lowest and highest in homozygotes with diplotypes 3 and 4 (130.1 +/- 40.8 and 194.2 +/- 44.6 mg/dl, P < 0.001), respectively. PPARD haplotype 3 was also an independent determinant of plasma apolipoprotein (apo)B (P = 0.021) and apoC-III (P = 0.001) levels, mean number of coronary lesions (P = 0.046) and changes in triglyceride (P = 0.01) and apoC-III (P = 0.047) levels in response to fluvastatin. Plasma triglyceride levels (P = 0.044), the mean number of coronary lesions (P = 0.026) and changes in minimum lumen diameter in response to fluvastatin (P = 0.022) were also associated with PPARG haplotypes. No significant associations between PPARA haplotypes and the phenotypes or significant interactions between PPAR haplotypes and the occurrence of new clinical events were detected. PPARD and PPARG haplotypes are independent determinants of plasma levels of lipids, severity of coronary atherosclerosis and its response to therapy. PMID- 15128053 TI - Association between Helicobacter pylori and recurrent aphthous stomatitis in children and adolescents. AB - BACKGROUND: The recurrent aphthous stomatitis (RAS) is a common disease with unknown etiology. Helicobacter pylori has been shown to be the causative factor in peptic ulcers. Considering the similarities of histologic features between gastric and oral ulceration, we studied the possible involvement of H. pylori in the development of RAS. METHODS: A total of 105 children and adolescents were investigated--53 patients with RAS (case group) and 52 patients without lesions (control group). Specimens obtained by swabbing RAS lesions, intact oral mucosa, and dental plaque were submitted to a polymerase chain reaction (PCR)-based assay. RESULTS: Helicobacter pylori was present in six patients of the case group (11.3%) and in three of the control group (5.8%). When the site of infection was studied, 9.4% of the RAS lesions were PCR positives. In the case group and control group, 5.7 and 1.9% of the specimens from dental plaque, respectively, and 5.7 and 3.8% of the specimens from the intact oral mucosa, respectively, were PCR positives. CONCLUSION: There was no association between RAS lesions and infection of the oral cavity by H. pylori in children and adolescents (P = 0.254). PMID- 15128054 TI - Serum interleukin-8 level is a more sensitive marker than serum interleukin-6 level in monitoring the disease activity of recurrent aphthous ulcerations. AB - BACKGROUND: Recurrent aphthous ulcerations (RAU) are common oral inflammatory lesions. Interleukin (IL)-8 is a pro-inflammatory cytokine of host response to injury and inflammation. Our recent study has found that measurement of serum IL 6 level can detect only 24% RAU patients with an abnormal serum level. In this study, we examined both the serum IL-6 and IL-8 levels in a group of RAU patients. The abilities of IL-6 and IL-8 to detect patients with an abnormal serum level were compared in order to find out whether IL-8 was a more sensitive serum marker than IL-6 in monitoring the disease activity of RAU. METHODS: In this study, we used a solid-phase, two-site sequential chemiluminescent immunometric assay to determine the baseline serum levels of IL-6 and IL-8 in 146 patients with RAU, 9 patients with traumatic ulcers (TU), and 54 normal control (NC) subjects. Eighty-two RAU patients, with the serum IL-6 or IL-8 levels higher than the upper limit of normal serum concentration, were treated with levamisole for 0.5-3.5 months, and their serum IL-6 and IL-8 levels were measured after treatment. RESULTS: We found that 25% (37/146) RAU patients, as well as 33% (20/61) major-type, 19% (13/69) minor-type, and 25% (4/16) herpetiform-type RAU patients, had a serum level of IL-6 greater than the upper normal limit of 4.7 pg/ml. In contrast, 60% (87/146) RAU patients, as well as 59% (36/61) major-type, 59% (41/69) minor-type, and 63% (10/16) herpetiform-type RAU patients, had a serum level of IL-8 greater than the upper normal limit of 8.7 pg/ml. In 82 RAU patients with the serum IL-6 or IL-8 levels higher than the upper limit of normal serum concentration, treatment with levamisole for a period of 0.5-3.5 months could significantly reduce the serum IL-6 level from 12.0 +/- 1.6 to 3.0 +/- 0.5 pg/ml (P < 0.001), and could significantly lower the serum IL-8 level from 70.9 +/- 11.2 to 13.8 +/- 3.1 pg/ml (P < 0.001). CONCLUSIONS: Because measurement of serum IL-8 level can detect 60% RAU patients with an abnormal serum level, while measurement of serum IL-6 level can detect only 25% RAU patients with an abnormal serum level, we conclude that serum IL-8 level is a more sensitive marker than serum IL-6 level in monitoring the disease activity of RAU. Levamisole can modulate both the serum IL-6 and IL-8 levels in RAU patients. IL-8, like IL-6, is also a useful serum marker in evaluating therapeutic effects of levamisole on RAU patients. PMID- 15128055 TI - The Th1 /Th2 immune-type response of the recurrent aphthous ulceration analyzed by cDNA microarray. AB - BACKGROUND: The reduced ability to activate oral tolerance plays a role in the pathogenesis of some gastrointestinal inflammatory diseases. This activation may reflect a preferential reduction of a T-helper (Th)2- or Th3-type response. In recurrent aphthous ulceration (RAU), genetic and environmental factors may contribute to low tolerance, permitting a cytotoxic reaction against the oral epithelium. The cytokine profile has not permitted the definition of RAU as resulting from enhanced Th1 or Th2 responses. A cDNA microarray study would allow the identification of differentially expressed genes and provide a basis for classification of the immune response. METHODS: The cDNA from 29 samples of aphthae and from 11 samples of normal mucosa from aphthae-free volunteers were hybridized on microarray membranes with 1176 genes. RESULTS: Forty-one differentially expressed genes were identified, and a higher expression level of the Th1 gene cluster in RAU was found. CONCLUSIONS: Microarrays permitted us definition of the gene expression profile of the lesion and identify an increased Th1 activity in RAU lesions. PMID- 15128056 TI - The inflammatory paradental cyst: a critical review of 342 cases from a literature survey, including 17 new cases from the author's files. AB - BACKGROUND: A total of 325 cases of inflammatory paradental cysts (IPCs) and 17 own cases were reviewed. Although known since 1930, the IPC is still unrecognized by many clinicians. The IPCs show a relative frequency of 0.9-4.7%. The majority of cysts occur distally or distobuccally to vital, permanent mandibular molars with a history of pericoronitis (IPC/3rd mandibular molar alone accounts for 64.9%). Radiologically, the cyst appears as a well-defined, semilunar unilocular radiolucency. MATERIALS AND METHODS: Cases of inflammatory paradental cysts and related lesions were retrieved from a worldwide literature survey. In addition, 17 new cases from the files of the authors have been added. RESULTS: The mean ages for patients with IPC/1st, 2nd and 3rd mandibular molars are 8.7, 17.4 and 27.6 years, respectively. The male:female ratio was 1 : 0.9 for IPC/1st and 2nd mandibular molars, and 1 : 0.4 for 3rd mandibular molar. CONCLUSION: Reduced enamel epithelium, cell rests of Malassez and remnants of the dental lamina stimulated by inflammation are thought to play a role in the pathogenesis of IPC. Histological features are indistinguishable from those of the inflammatory, periapical (radicular) cyst. PMID- 15128057 TI - Immunohistochemical localization of tissue-type plasminogen activator and type I plasminogen activator inhibitor in radicular cysts. AB - BACKGROUND: The plasminogen/plasmin proteolytic system participates in a wide variety of extracellular matrix degradation. Detailed knowledge of plasminogen activators (PAs) and their inhibitors may be important for understanding the pathogenesis of radicular cysts. The purpose of this study was to investigate the in situ localization of tissue-type PA (t-PA) and type I PA inhibitor (PAI-1) in radicular cysts. METHODS: Thirty formalin-fixed, paraffin-embedded specimens of radicular cysts were examined using immunohistochemistry. In addition, another section from each radicular cyst specimen was stained with hematoxylin and eosin to assess the presence of inflammatory infiltrates. Differences in t-PA and PAI-1 expression between tissues with low and high levels of inflammation were subsequently analyzed using Fisher's exact test. RESULTS: Both t-PA- and PAI-1 positive cells were detected in the lining epithelium, connective tissue, inflammatory infiltrates, and endothelium. In addition, the t-PA signal was mainly expressed in epithelial cells. However, the PAI-1 signal was mainly expressed in fibroblasts. Moreover, significantly greater t-PA as well as PAI-1 expression was noted in radicular cysts with high levels of inflammation as compared to tissues with low levels of inflammatory cell infiltrates (P < 0.05). CONCLUSIONS: The present study confirms earlier indications of local production of PA and its inhibitor in radicular cysts. In addition, this study further shows the tissue localization of the antigens for t-PA as well as PAI-1, and demonstrates that the expression of both t-PA and PAI-1 increases with the grade of inflammation in radicular cysts. PMID- 15128058 TI - The role of lipopolysaccharide in infectious bone resorption of periapical lesion. AB - BACKGROUND: The role of lipopolysaccharide (LPS) in periapical lesion-induced bone resorption was investigated. Polymyxin B (PMB), a specific inhibitor of LPS, was evaluated to treat the apical lesion. METHODS: Lipopolysaccharide isolated from two common endodontic pathogens, Fusobacterium nucleatum and Porphyromonas endodontalis, stimulated mouse macrophage (J774) to release interleukin-1alpha (IL-1 alpha) and tumor necrosis factor-alpha (TNF-alpha) in a time-dependent manner. RESULTS: Combination of LPS further enhanced the stimulation. PMB inhibited these effects significantly. LPS also stimulated matrix metalloproteinase-1 (MMP-1) gene expression in J774, whereas anti-IL-1 alpha and anti-TNF-alpha antibodies, as well as PMB, diminished this effect. A disease model of periapical lesion was established in Wistar rat. Administration of PMB reduced the extent of lesion-associated bone resorption by 76% to approximately 80%, and simultaneously reduced the numbers of MMP-1-producing macrophages. CONCLUSIONS: It is suggested that LPS released from the infected root canal triggers the synthesis of IL-1 alpha and TNF-alpha from macrophages. These pro inflammatory cytokines up-regulate the production of MMP-1 by macrophages to promote periapical bone resorption. PMID- 15128059 TI - Central giant cell granuloma of the jaws: assessment of cell cycle proteins. AB - BACKGROUND: Several reports have demonstrated the presence of a high proliferative activity in central giant cell granuloma, raising the possibility that deregulation of the cell cycle may contribute to its pathogenesis. As we identified alterations of cyclin D1 in giant cell tumor of bone, and as there are histologic similarities between central giant cell granuloma and giant cell tumor, we assessed jaw lesions for the presence of similar alterations. METHODS: Formalin-fixed, paraffin-embedded tissue from 29 cases of central giant cell granuloma was assessed for the expression of cyclin D1, cyclin B1, and MIB-1 (Ki 67) using immunohistochemistry. In addition, differential polymerase chain reaction (PCR) was used to determine whether there was cyclin D1 gene amplification. RESULTS: The cyclin D1 gene copy number appeared to be minimally elevated in 31% of the cases. Cyclin D1 protein overexpression was observed in 28 of 29 cases (96.5%). Immunostaining was present predominantly in the nuclei of the giant cells. Cyclin B1 and MIB-1 immunoreactivity was restricted to the mononuclear cells with no staining present in the giant cells. CONCLUSIONS: Cyclin D1 protein overexpression may be involved in the formation of the giant cells and the pathogenesis of central giant cell granuloma. As the distribution of immunostaining is identical to that observed in giant cell tumor of bone, our results support the possibility that central giant cell granuloma of the jaws and giant cell tumor of bone represent a similar disease process that clinically and histologically may have somewhat different features because of differences in the anatomical site of involvement. PMID- 15128060 TI - Basement membrane-type heparan sulfate proteoglycan (perlecan) and low-density lipoprotein (LDL) are co-localized in granulation tissues: a possible pathogenesis of cholesterol granulomas in jaw cysts. AB - BACKGROUND: As perlecan contains a low-density lipoprotein (LDL) receptor-like repeats in the second domain of its core protein, LDL may be bound to perlecan, which is rich in granulation tissues. We wanted to study if this is the case in the cyst wall of radicular cysts, which are often associated with cholesterol granuloma. METHODS: Thirty-three specimens of radicular cyst with cholesterol granulomas were immunohistochemically examined for comparative localizations of perlecan, apoprotein B (apo B), and oxidized LDL (Ox-LDL), and for mRNA expression levels for perlecan. RESULTS: Myxoid or edematous stroma of immature granulation tissues was strongly positive for perlecan and simultaneously for apo B and Ox-LDL. Macrophages including foamy cells scattered in the granulation tissues were also immunopositive for Ox-LDL and occasionally for apo B. In situ hybridization showed that fibroblasts, endothelial cells, and pericytes had strong signals for perlecan, which was also confirmed by RT-PCR. CONCLUSION: These results suggest that perlecan, which is abundantly produced and accumulated in the cyst wall of immature granulation tissue, traps Ox-LDL locally, and that Ox-LDL is phagocytosed by macrophages. Thus, LDL-laden foamy macrophages are aggregated in the granulation tissue, and free cholesterol from ruptured macrophages may be concentrated locally to be crystallized, which may induce foreign body granulomas in the cyst wall. PMID- 15128061 TI - Expression of Sonic hedgehog (SHH) signaling molecules in ameloblastomas. AB - BACKGROUND: To clarify the roles of Sonic hedgehog (SHH) signal transduction in oncogenesis and cytodifferentiation of odontogenic tumors, expression of SHH, Patched (PTC), Smoothened (SMO), and GLI1 was analyzed in ameloblastomas as well as in tooth germs. METHODS: Tissue specimens of 9 tooth germs, 36 benign ameloblastomas, and 1 malignant ameloblastoma were examined by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry for the expression of SHH, PTC, SMO, and GLI1. RESULTS: Expression of SHH, PTC, SMO, and GLI1 mRNA was detected in all tooth germ and ameloblastoma samples. Immunohistochemical reactivity for SHH, PTC, SMO, and GLI1 was detected in both normal and neoplastic odontogenic tissues. Expression of SHH, PTC, and GLI1 was more evident in epithelial cells than in mesenchymal cells, whereas SMO reactivity was marked in both epithelial and mesenchymal components in tooth germs and ameloblastomas. In ameloblastomas, these SHH signaling molecules were expressed more intensely in peripheral columnar or cuboidal cells than in central polyhedral cells; keratinizing cells and granular cells showed no or little reactivity. CONCLUSION: Expression of SHH, PTC, SMO, and GLI1 in tooth germs and ameloblastomas suggests that these SHH signaling molecules might play a role in epithelial-mesenchymal interactions and cell proliferation in tooth development as well as in growth of these epithelial odontogenic tumors. PMID- 15128062 TI - Does acute alcoholic pancreatitis exist with preexisting chronic pancreatitis? PMID- 15128063 TI - Alcoholic pancreatitis: is it a priori chronic disease? PMID- 15128064 TI - Ethics of sample collecting in gastrointestinal endoscopy-based research. PMID- 15128065 TI - Endoluminal therapies for gastroesophageal reflux disease. AB - While medical therapy, particularly with proton pump inhibitors, is effective for the large majority of patients with reflux disease, there remains a subset of patients who are dissatisfied, due to cost, side effects of medications, or persistent symptoms such as regurgitation. For this population, surgical fundoplication has been, and remains, an appropriate option. A new class of endoluminal interventions, attempting to create a mechanical antireflux barrier, has emerged recently. Three such devices are currently approved and available, and a number of others are in various stages of evaluation. This article will review the approved technologies, as well as selected promising emerging ones. with particular emphasis on the scientific evidence available to date supporting their efficacy. PMID- 15128066 TI - Metabolic bone disease in inflammatory bowel disease. AB - Osteoporosis is associated with a high morbidity rate and is a risk factor for fractures. Patients with inflammatory bowel disease are at increased risk of developing osteopenia and osteoporosis. Corticosteroid use, malnutrition, and proinflammatory cytokines are unique risk factors for bone loss in ulcerative colitis and Crohn disease. Bone mineral density is assessed by dual-energy X-ray absorptiometry and reported as a T score or number of standard deviations away from the mean. A T score < 1 SD below the mean is normal, 1 to 2.5 SD below the mean is consistent with osteopenia, and greater than 2.5 SD below the mean is defined as osteoporosis. Treatment includes a combination of basic preventative measures, for example, weight-bearing exercise, calcium, vitamin D, and pharmacologic agents, (e.g., bisphosphonates). PMID- 15128067 TI - The need for caution with topical anesthesia during endoscopic procedures, as liberal use may result in methemoglobinemia. AB - During upper gastrointestinal endoscopy, topical oropharyngeal anesthesia with lidocaine and/or benzocaine is used routinely by many endodscopists. Although such a practice is usually safe, there have been a number of reports of methemoglobinemia induced by topical anesthesia. Early treatment is extremely important as the development of methemoglobinemia is potentially fatal. Methemoglobinemia should be considered when oxygen desaturation occurs without another explanation. In this case series, we report 4 cases of methemoglobinemia that followed the liberal application of Cetacaine for ERCP. All patients recovered after appropriate treatment but these cases serve to highlight the potential problem, the importance of early recognition and treatment, and the most appropriate treatment options. PMID- 15128068 TI - Gastric emptying and intragastric distribution of a solid meal in functional dyspepsia: influence of gender and anxiety. AB - BACKGROUND: Gastric dysmotility and psychological factors have been implicated in the pathophysiology of functional dyspepsia. GOALS: To investigate the influence of gender and anxiety on gastric emptying and intragastric food distribution in patients with functional dyspepsia. METHODS: A standard gastric emptying test was used to study total and compartmental gastric emptying of a solid meal in 22 patients with functional dyspepsia (16 women). Comparisons of the data for dyspeptic men and women were made with 2 respective subgroups of controls (9 men; 9 women). The presence of anxiety and depression was assessed by the Hospital Anxiety and Depression scale. RESULTS: Delayed gastric emptying was observed in 32% of patients with functional dyspepsia. As a group, dyspeptic women had a significantly longer half-emptying time as compared with dyspeptic men ( 19 +/- 41 min vs. 78 +/- 22 min) and to female controls (96 +/- 17 min). There was no difference in half-emptying times between male patients and controls. The initial activity in the proximal stomach was significantly lower for both men and women with functional dyspepsia in comparison with their respective controls. In addition, meal retention in the distal stomach of dyspeptic women was significantly greater than that in female controls. Sixteen (72%) functional dyspepsia patients had anxiety when evaluated by the Hospital Anxiety and Depression scale. Those patients who scored for anxiety showed significantly greater antral meal retention than patients without anxiety. CONCLUSION: Our study confirmed that prolonged gastric emptying in patients with functional dyspepsia is related to the female sex, while the abnormalities of the meal intragastric distribution appear to occur in dyspeptic males and females. Anxiety is frequent in functional dyspepsia and seems to be related to abnormal antral retention of food in these patients. PMID- 15128069 TI - Erythromycin in the short- and long-term control of dyspepsia symptoms in patients with gastroparesis. AB - BACKGROUND: Few prokinetic drugs are available to treat gastroparesis. Data are limited on short-term and long-term efficacy of erythromycin as a prokinetic drug. GOALS: Assess efficacy of low-dose erythromycin suspension to treat gastroparesis. STUDY: Patients with dyspepsia and gastroparesis by gastric emptying study were treated with low-bulk diet and low-dose (50-100 mg 3 times a day and at bedtime) oral erythromycin suspension. Data were collected by retrospective chart review and telephone questionnaire for short- and long-term follow-up, respectively. RESULTS: Of 25 patients, 18 had short-term follow-up, 18 had longterm follow-up, and 14 had both. On short-term follow-up, 15 patients (83%) experienced some or dramatic improvement, while 3 (17%) experienced worsening or no change in symptoms (P = 0.005). Mean duration of long-term use was 11 +/- 7 months. On long-term followup, 12 (67%) patients noticed some or dramatic improvement, while 6 (33%) experienced worsening or no change in symptoms (P = 0.16). Correlation (0.7) between short- and long-term response was significant (P < 0.005). Of the 3 patients with poor short-term response, none did well long term. Of the 11 patients with some or dramatic response in short term, 7 continued to have some response long term. There was no relation between gastric emptying time and response to erythromycin suspension. CONCLUSIONS: Treatment of gastroparesis with low-dose erythromycin and low-bulk diet results in a dramatic short-term improvement in the majority of patients. Short-term response predicts long-term response. This response may not be as great, possibly due to tachyphylaxis. PMID- 15128070 TI - The effect of octreotide as an adjunct treatment in active nonvariceal upper gastrointestinal bleeding. AB - GOALS: The aim of this study was to determine the effect ofoctreotide on active or recent gastrointestinal bleeding from benign peptic ulcers. STUDY: This is a prospective, randomized study including 110 patients with gastric or duodenal peptic ulcers presenting with active spurting or oozing bleeding or nonbleeding visible vessel. All patients were subjected to endoscopic hemostasis by injection of noradrenaline, and they were then randomized to either receive octreotide (55 patients) or placebo (55 patients). The groups did not differ with respect to age, sex, use of nonsteroidal antiinflammatory drugs, previous history of ulcer or bleeding, Helicobacter pylori infection, site, and severity of bleeding. RESULTS: The rebleeding rate was 36% in placebo and 32% in octreotide group, which does not present a statistically significant difference. Surgical intervention was required for 18 patients (32.7%) in the placebo group and for 16 patients (29%) in the octreotide group. The mortality rate was 2 patients (3.6%) in the placebo and 4 patients (7.2%) in the octreotide group. All the above presented no statistical difference. In addition, there was no statistically significant difference between the 2 groups with respect to the number of blood units transfused and hospital stay. CONCLUSIONS: The use of octreotide as an adjunct treatment in patients with acutely bleeding benign peptic ulcer or/and visible vessel did not seem to offer significant benefits regarding their outcome. PMID- 15128071 TI - Ideology of Helicobacter pylori prevalence in peptic ulcer disease in an inner city minority population. AB - GOALS AND BACKGROUND: The prevalence of Helicobacter pylori infection among patients with peptic ulcer disease has been reported to range from 61 to 94%. Recent studies show a reduction in the prevalence of H. pylori infection in patients with peptic ulcer disease. This study was conducted to determine the prevalence of H. pylori infection in peptic ulcer disease in an inner-city hospital in Washington, DC. METHODS: Medical records of all patients who had undergone upper gastrointestinal endoscopy from July 1997 through June 1999 were reviewed. All patients who had gastric ulcer and/or duodenal ulcer on upper gastrointestinal endoscopy were studied. Demographic characteristics, history of nonsteroidal antiinflammatory drug ingestion, alcohol consumption, and associated diseases were studied. H. pylori was considered to be present if CLOtest and/or histopathology were positive for H. pylori. Patients with negative pathology for H. pylori or negative pathology and CLOtest were considered negative for H. pylori. RESULTS: One-hundred fifty-six patients were found to have gastric and/or duodenal ulcers. Fifty-one ulcer patients did not meet the inclusion criteria and were excluded. Among the 105 patients who were included in the study, gastric ulcers were found in 48 patients (45.7%), duodenal ulcers were found in 46 patients (43.8%), and both gastric and duodenal ulcers were found in 11 patients (10.5%). H. pylori was present in 66.7% of gastric ulcer patients and in 69.5% of duodenal ulcer patients. Antral histology and CLOtest were in agreement 96% of the time. CONCLUSIONS: At the District of Columbia General Hospital, an inner city hospital serving predominantly an African-American community, the prevalence of H. pylori in ulcer patients compares similarly to other more recent studies that have found a decreased prevalence of this bacterial infection in ulcer patients. This suggests that the treatment of H. pylori in minority patients is reducing the proportion of ulcers due to this bacterium, as has been seen with the majority population. PMID- 15128072 TI - Abnormal expression of E-cadherin in early gastric carcinoma: its relationship with macroscopic growth patterns and catenin alpha and beta. AB - BACKGROUND: Abnormal expression of E-cadherin plays an important role in the differentiation and progression of gastric carcinoma. There are a few identifiable clinical, histopathologic, and molecular changes that are related with the macroscopic growth patterns of gastric carcinoma. The aim of this study is to elucidate the relation between the macroscopic growth patterns of gastric carcinoma and the abnormal expression of E-cadherin, alpha- and beta-catenins. STUDY: A total of 97 cases of early gastric carcinoma were examined by immunohistochemistry using monoclonal antibodies against E-cadherin, alpha- and beta-catenins. Macroscopically, 52 cases were elevated types and 45 cases were depressed types. RESULTS: Early gastric carcinomas with depressed growth showed diffuse histologic type, younger patients' age and smaller size more frequently than tumors with elevated growth. Abnormal expression rates of E-cadherin, alpha- and beta-catenins were 35.1, 36.1, and 46.4%, respectively. Abnormal expression of E-cadherin, alpha- and beta-catenins was significantly associated with depressed tumor growth and diffuse histologic type. By multiple logistic regression analysis. E-cadherin, age, Lauren classification, World Health Organization grade and size were identified as risk factors of macroscopic growth pattern of early gastric carcinomas. Positive associations between E-cadherin and beta-catenin and between alpha- and beta-catenins were found by log linear model analysis. CONCLUSION: We suggest that E-cadherin plays an important role in the macroscopic growth as well as microscopic differentiation of early gastric carcinomas. PMID- 15128073 TI - Diagnosis and treatment of postprandial nausea, allergy, and eosinophilia. AB - We report a case of a 16-year-old male patient who presented with postprandial fullness and nausea. He had a history of seasonal allergies, asthma, and peripheral eosinophilia. Endoscopy of the stomach with mucosal biopsies revealed predominate eosinophils. A diagnosis of eosinophilic gastroenteritis was made. The patient's disease course and management is described in this article. PMID- 15128074 TI - Epidemiology and pathogenesis of primary biliary cirrhosis. AB - Primary biliary cirrhosis is an autoimmune disease of unknown etiology leading to progressive destruction of small intrahepatic bile ducts and eventually to liver cirrhosis and failure. It is characterized by female predominance (with most cases observed between the ages of 40 and 60) and serum autoantibodies to mitochondrial antigens as highly specific hallmarks. Epidemiologic data indicate a variable incidence and prevalence of the disease. A number of genetic factors have been indicated as playing a role in determining disease susceptibility or progression, although no definitive conclusion has been reached so far. However, as suggested by some epidemiologic observations, a number of environmental factors, including molecular mimicry by either microorganisms or xenobiotics, have also been proposed. A hypothesis gaining support is that environmental factors may trigger disease in genetically predisposed individuals. In this review, the available data regarding the epidemiology and pathogenesis of primary biliary cirrhosis will be described and discussed. PMID- 15128075 TI - Does acute alcoholic pancreatitis precede the chronic form or is the opposite true? A histological study. AB - OBJECTIVES: Whether acute alcoholic pancreatitis occurs in a normal pancreas or in a pancreas that has already been altered by chronic pancreatitis is unclear. Our objective is to clarify the relation between acute and chronic alcoholic pancreatitis by histologic study of the pancreas in a group of patients having a first attack of acute alcoholic pancreatitis. METHODS: From January 1989 to December 1999, 138 patients with acute pancreatitis, of whom 28 had alcoholic pancreatitis, were seen by us; in 21 of the latter 28 patients, it was the first attack. Of these 21, 6 underwent surgery for acute necrotic pancreatitis. In all 6 patients, adequate pancreatic biopsies were obtained during surgery. Tissue samples were prepared for histologic examination according to standard procedures. RESULTS: In all 6 patients, both acute necrotic and chronic lesions were found. The chronic lesions had characteristics of chronic calcifying pancreatitis and consisted of perilobular and intralobular fibrosis. loss of exocrine parenchyma, dilated interlobular ducts, and protein plugs within dilated ducts. CONCLUSIONS: This study suggests that acute alcoholic pancreatitis develops in a pancreas already affected by chronic pancreatitis. The hypothesis that in alcoholics chronic pancreatitis derives from acute pancreatitis is not supported by the present data. PMID- 15128076 TI - Risk of acute pancreatitis in users of finasteride: a population-based case control study. AB - GOALS: To examine the risk of acute pancreatitis in > or = 60-year-old male users of finasteride in a case-control study from a population of 490,000 persons in North Jutland County, Denmark, from 1993 to 2000. STUDY: We identified all men age 60 and over with incident acute pancreatitis from the County Hospital Discharge Registry in North Jutland, 1993 to 2000, and selected 10 age-matched controls per case from the Danish Civil Registration System based on incidence density sampling. All prescriptions for finasteride within 90 days prior to admission with acute pancreatitis were identified from the population-based North Jutland Prescription Database. Data on potential confounding factors were also extracted from registries. We used conditional logistic regression to estimate the relative risk of acute pancreatitis, adjusted for these potential confounders. RESULTS: There were 302 men age 60 and older with incident acute pancreatitis, of whom 3 were exposed to finasteride, and 2994 controls, of whom 37 were exposed to finasteride. The crude odds ratio for having reimbursed prescriptions for finasteride was 0.8 (95% confidence interval, 0.2-2.6). After adjustment for alcohol-related diseases, gallstone disease, hyperlipidemia, hypercalcemia, and hyperparathyroidism, the odds ratio was slightly decreased to 0.5 (95% confidence interval, 0.1-2.5). CONCLUSION: We did not find any increased risk of acute pancreatitis in users of finasteride. PMID- 15128077 TI - Hepatitis C tested prevalence and comorbidities among veterans in the US Northwest. AB - GOALS: (1) Investigate the epidemiology of hepatitis C virus infection among patients seen in the Veterans Administration Northwest Network; (2) examine time trends in testing practices and results; and (3) estimate the prevalence of hepatitis C virus infection among active patients. BACKGROUND: Hepatitis C virus infection causes chronic hepatitis and cirrhosis and is a leading cause of end stage liver disease. Hepatitis C virus antibodies are estimated to be present in 1.8% of the US population, but reports of its prevalence among US veterans range from 1.7 to 35%. STUDY: Retrospective review of computerized medical records of veterans tested for hepatitis C from October 1994 through December 2000 (n = 37,938) at 8 Northwest Veterans Administration Medical Centers. RESULTS: Among tested veterans, 8230 (21.7%) had evidence of hepatitis C virus infection. The number of patients tested increased annually from 2335 to 18,191, while the proportion with first-time positive hepatitis C test results decreased from 35 to 10%. This drop in tested prevalence was associated with a shift away from testing individuals at highest risk--those with positive hepatitis B serostatus, repeatedly elevated alanine transaminase levels, and drug use disorder diagnoses. We estimate that 11.4% of the Northwest Network veteran users are hepatitis C virus seropositive, with a lower bound of 4.0% and upper bound of 19.5%. CONCLUSIONS: Although estimates of hepatitis C virus infection rates among veteran users of the Veterans Administration system remain higher than those for the general population, changes in testing practice make generalizations from earlier studies hazardous. PMID- 15128078 TI - Hepatobiliary fascioliasis: clinical and radiologic features and endoscopic management. AB - Fasciola hepatica is a zoonotic liver fluke that can cause disease in humans. Fascioliasis is an uncommon disease. We retrospectively analyzed 9 cases of fascioliasis and reviewed the relevant literature. A high index of suspicion and specific ultrasonographic findings are very helpful in the diagnosis of the disease. However, serological studies and endoscopic retrograde cholangiopancreatography confirm the diagnosis. The disease has 2 stages: hepatic stage and biliary stage. While several drugs are used during the hepatic stage, endoscopic retrograde cholangiopancreatography is particularly effective in the biliary stage. PMID- 15128079 TI - Liver transplantation for disulfiram-induced fulminant hepatic failure. AB - Disulfiram has been used as a popular adjunct in programs for alcohol rehabilitation. However, in rare cases, disulfiram has been reported to cause fulminant hepatitis. Disulfiram use and its associated complications in adolescents have received minimal attention in the literature. We report the first pediatric case of successful orthotopic liver transplantation for disulfiram-induced fulminant hepatic failure in a 16-year-old girl, who developed an idiosyncratic reaction associated with short-term, low-dose disulfiram use, as evidenced by her liver biopsy and explanted liver. This case report indicates that a high index of suspicion, and aggressive therapeutic interventions are necessary to recognize and manage disulfiram-induced fulminant hepatic failure in adolescents. Success of this case suggests that transplant centers should consider liver transplantation of an adolescent alcoholic patient with fulminant hepatic failure due to non-alcohol-related causes such as disulfiram. Orthotopic liver transplantation should be considered early in the management of disulfiram induced fulminant hepatic failure. PMID- 15128080 TI - Hepatocellular carcinoma containing sarcomatous lesions in a normal liver, accompanied by secondary Budd-Chiari syndrome. PMID- 15128081 TI - Administration of H2 receptor antagonist with proton pump inhibitor is effective for long-term control of refractory reflux esophagitis. PMID- 15128082 TI - Gastric cancer presenting as gastropericardial fistula in a patient with familial adenomatous polyposis syndrome. PMID- 15128083 TI - An examination of the transmissibility and clinical utility of auscultation of bowel sounds in all four abdominal quadrants. PMID- 15128084 TI - Methicillin-resistant Staphylococcus aureus as a cause of antibiotic-associated diarrhea. PMID- 15128085 TI - Cat scratch disease presenting with abdominal pain and retroperitoneal lymphadenopathy. PMID- 15128086 TI - Soluble interleukin 2 receptor plasma levels in non-responders with chronic hepatitis C treated with pegylated interferon-alpha 2b and ribavirin. PMID- 15128087 TI - Cure of acute hepatitis C in HIV co-infection? PMID- 15128088 TI - The patient at the center of care. PMID- 15128089 TI - What nursing interventions reduce the risk of symptomatic urinary tract infection in the patient with an indwelling catheter? PMID- 15128090 TI - Reading and using systematic reviews. PMID- 15128091 TI - Antiphospholipid antibody thrombosis. Another source of chronic wounds. AB - Antiphospholipid antibody thrombosis syndrome is currently not well known and probably underdiagnosed. The risk it presents for recurrent thromboembolism is both life threatening and chronic. Because patients often develop long-standing wounds, it is important for the wound care nurse to identify clients suspect for the disease. Several theories exist speculating on the mechanism of thrombosis involving antiphospholipid antibodies, and each focuses on changes at the interface between the vessel, cell wall membrane, and its interaction with circulating antibodies. As wound care nurses, we should consider antiphospholipid antibody thromboses whenever encountering a recalcitrant wound without obvious underlying cause. Thrombosis can occur anywhere, although 41% of presentations are cutaneous. Recurrent deep vein thrombosis with coexisting venous insufficiency can lead to a missed diagnosis of antiphospholipid thrombosis syndrome, despite evidence of pain and tissue necrosis that is typical of the disease. In any event, long-standing nonhealing wounds are always a signal to revise a diagnosis and seek alternative causes for failure of healing. PMID- 15128092 TI - Cecostomy for antegrade continence enemas in children. PMID- 15128093 TI - A multicenter comparative evaluation: disposable pads for women with light incontinence. AB - OBJECTIVE: The purpose of this study was to evaluate the current range of disposable pads for women with light incontinence available in the United Kingdom. SETTING AND SUBJECTS: Sixty community-based women aged 50 years or older were recruited to the study from several locations throughout the United Kingdom. METHODS: A multiple crossover design was used, which enabled all subjects to test all of the 12 products that were available on the UK market at the time of the evaluation. Two tools were used to evaluate the products: a product performance questionnaire and a pad leakage diary. This incorporated both subjective and objective outcome measures. RESULTS: Statistically significant differences were found between the products in 13 of the 17 performance aspects, eg, ability to hold urine without leaking and the fit of the pad. CONCLUSIONS: As a group, all of the products tested performed well in terms of their ability to hold urine without leakage. However, two products performed particularly well and scored highly on most aspects of performance and represented successful "all-round" products; these were the Tena Lady Extra and the Prevail Extra Plus. PMID- 15128094 TI - Evaluation of an evidence-based education program for pressure ulcer prevention. AB - OBJECTIVE: The purpose of this study was to implement and evaluate a standardized education workshop for 2 levels of nursing staff. DESIGN: The quasi-experimental design required that nurses' knowledge be measured before and after education. SETTING AND SUBJECTS: The convenience sample included registered nurses (N = 595) and licensed practical nurses (N = 59) employed in three acute care hospitals with a total bed capacity of 1,760. INSTRUMENTS: The Pressure Ulcer Knowledge Test, including true-false questions, was selected and compared with updated literature and the evidence-based standards developed for the acute care hospitals. Minor changes resulted in a revised 53-item questionnaire for which content validity and reliability were established. The questionnaire was pilot tested before use. METHODS: The intervention, the standardized education workshop, included risk assessment, prevention, and wound staging, as described in the literature. The dependent variable, knowledge of pressure ulcer prevention and treatment, was measured at 3 time points: immediately before (pre) and after (post 1) the workshop and 3 months later (post 2). Data were analyzed using descriptive statistics and repeated measures. RESULTS: Overall, knowledge scores for the total group were significantly higher from pre to post 1 and from pre to post 2 but significantly lower from post 1 to post 2. Registered nurses' scores were significantly higher than those of licensed practical nurses from pre to post 1. CONCLUSION: The evidence-based pressure ulcer education was effective in increasing registered nurses' and licensed practical nurses' knowledge and was presumed to increase consistency of the knowledge and awareness of practice standards. PMID- 15128095 TI - Preventing and managing perineal dermatitis: a shared goal for wound and continence care. AB - Perineal dermatitis is frequently encountered in patients with urinary and/or fecal incontinence, particularly when managed by a containment device such as a pad or containment brief. In addition to intervening to prevent or alleviate incontinence, a preventive perineal skin program in the high-risk patient and aggressive management of incontinence-related dermatitis are both essential for effective holistic management. A routine cleansing regimen that removes irritants and moisturizes while protecting the skin from additional irritation is effective for prevention of dermatitis and for managing mild cases. However, more aggressive care is required when perineal dermatitis occurs, especially when it is associated with epidermal erosion. Xenaderm ointment combines three active ingredients in a hydrophobic ointment that promotes healing while acting as a moisture barrier to prevent further irritation and damage. PMID- 15128096 TI - DNA detection by integrable electronics. AB - This paper presents a new electronic methodology to detect DNA hybridization for rapid identification of diseases, as well as food and environmental monitoring on a genetic base. The proposed solution exploits a new (electrical) capacitive measurement circuit, not requiring any prior labeling of the DNA (as it is often the case with the commonly employed optical detection). The sensitivity, the reliability, and the reproducibility of this device have been evaluated by experiments performed with a (non-integrated) prototype implementation, easily integrable in IC and/or micro-fabricated lab-on-a-chip. PMID- 15128097 TI - Prussian blue modified amperometric FIA biosensor: one-step immunoassay for alpha fetoprotein. AB - The aim of this study was to develop a rapid method to measure alpha-fetoprotein (AFP) in human serum by use of one-step sandwich enzyme-linked immunosorbent assay (ELISA)-based immunobiosensor with disposable screen-printed carbon electrode technology. Prussian blue (PB) was deposited using cyclic voltammetry (CV) on the surface of electrode to catalyze H202 from the reaction of glucose oxidase. It took only about 30 min to complete the whole experimental procedure through flow injection analysis (FIA). A detection range obtained is in the range from 5 to 500 ng/ml AFP. This detection seems to be quick, reliable and practical. PMID- 15128099 TI - Application of biochemical oxygen demand (BOD) biosensor for optimization of biological carbon and nitrogen removal from synthetic wastewater in a sequencing batch reactor system. AB - A bench scale reactor using a sequencing batch reactor process was used to evaluate the applicability of biosensors for the process optimization of biological carbon and nitrogen removal. A commercial biochemical oxygen demand (BOD) biosensor with a novel microbial membrane was used to determine the duration of each phase by measuring samples in real time in an SBR cycle with filling/anoxic-anaerobic/aerobic/sludge wasting/settling/withdrawal periods. Possible strategies to increase the efficiency for the biological removal of carbon and nitrogen from synthetic wastewater have been developed. The results show that application of a BOD biosensor enables estimation of organic carbon, in real time, allowing the optimization or reduction the SBR cycle time. Some typical consumption patterns for organic carbon in the non-aeration phase of a typical SBR operation were identified. The rate of decrease of BOD measured using a sensor BOD, was the highest in the initial glucose breakdown period and during denitrification. It then slowed down until a 'quiescent period' was observed, which may be considered as the commencement of the aeration period. Monitoring the BOD curve with a BOD biosensor allowed the reduction of the SBR cycle time, which leads to an increase in the removal efficiency. By reducing the cycle time from 8 to 4 h cycle, the removal efficiencies of nitrate, glucose, and phosphorus in a given time interval, were increased to nearly double, while the removal of nitrogen ammonium was increased by one-third. PMID- 15128098 TI - A chemiluminescence flow immunosensor based on a porous monolithic metacrylate and polyethylene composite disc modified with protein G. AB - A generic, fast, sensitive and new type of flow immunosensor has been developed. The basis is a monolithic porous poly(glycidyl methacrylate-co-trimethylolpropane trimethacrylate) polymer disc modified with protein G, placed in a fountain type flow cell compartment, in close proximity to a photomultiplier tube (PMT). Analyte and HRP labelled analyte derivative (tracer) compete for anti-analyte antibody binding sites. The mixture is then injected into the flow immunosensor system where the formed analyte- and tracer-antibody complexes are trapped by the monolithic protein G disc. The amount of bound tracer, inversely related to the concentration of analyte in the sample, is determined in a second step by injection of luminol, p-iodophenol and H2O2, generating enhanced chemiluminescence (CL) with horseradish peroxidase (HRP). A third and final step is need for regeneration of the protein G disc so that a new analysis cycle can take place. The performance of the disc immunosensor system was compared with a one step continuous flow injection immunoassay (FIIA) system, using the same reagents and a protein G column, in terms of assay sensitivity and influence of matrix effects from various water samples (millipore-, tap- and surface water). The detection limit for the analyte atrazine in PBS and surface water (SW) was 0.208 +/- 0.004 microg l(-1) (PBS) and 0.59 +/- 0.120 microg l(-1) (SW) for the FIIA and 0.033 +/- 0.003 microg l(-1) (PBS) and 0.038+/-0.003 microg l(-1) (SW) for the disc immunosensor. Statistical comparison of the two systems shows that the disc immunosensor results were significantly less influenced by the sample matrix, which is explained by the fact that the sample in the FIIA arrives simultaneously with the matrix to the detector, whereas these are separated in time in the disc immunosensor system. PMID- 15128100 TI - Comparative study of controlled pore glass, silica gel and poraver for the immobilization of urease to determine urea in a flow injection conductimetric biosensor system. AB - This study compared the responses of three enzyme reactors containing urease immobilized on three types of solid support, controlled pore glass (CPG), silica gel and Poraver. The evaluation of each enzyme reactor column was done in a flow injection conductimetric system. When urea in the sample solution passed though the enzyme reactor, urease catalysed the hydrolysis of urea into charged products. A lab-built conductivity meter was used to measure the increase in conductivity of the solution. The responses of the enzyme reactor column with urease immobilized on CPG and silica gel were similar and were much higher than that of Poraver. Both CPG and silica gel reactor columns gave the same limit of detection, 0.5 mM, and the response was still linear up to 150mM. The analysis time was 4-5 min per sample. The enzyme reactor column with urease immobilized on CPG gave a slightly better sensitivity, 4% higher than the reactor with silica gel. The life time of the immobilized urease on CPG and silica gel were more than 310h operation time (used intermittently over 7 months). Good agreement was obtained when urea concentrations of human serum samples determined by the flow injection conductimetric biosensor system was compared to the conventional methods (Fearon and Berthelot reactions). These were statistically shown using the regression line and Wilcoxon signed rank tests. The results showed that the reactor with urease immobilized on silica gel had the same efficiency as the reactor with urease immobilized on CPG. PMID- 15128101 TI - Enzymatic activity of glucose oxidase covalently wired via viologen to electrically conductive polypyrrole films. AB - The surface functionalization of an electrically conductive polypyrrole film (PPY) with a viologen, (N-(2-carboxyl-ethyl)-N'-(4-vinyl-benzyl)-4,4' bipyridinium dichloride, or CVV) for the covalent immobilization of glucose oxidase (GOD) has been carried out. The viologen was first synthesized and graft polymerized on PPY film. It then served as an anchor via its carboxyl groups for the covalent immobilization of GOD. The surface composition of the as functionalized substrates was characterized by X-ray photoelectron spectroscopy (XPS). The effects of the CVV monomer concentration on the CVV-graft polymer concentration and the amount of GOD immobilized on the surface were investigated. The activity of the immobilized GOD was compared with that of free GOD and the kinetic effects were also obtained. The cyclic voltammetric (CV) response of the GOD-functionalized PPY substrates was studied in a phosphate buffer solution under an argon atmosphere. The CV results support the mechanism in which CVV acts as a mediator to transfer electron between the electrode and enzyme, and hence regenerating the enzyme in the enzymatic reaction with glucose. High sensitivity and linear response of the enzyme electrode was observed with glucose concentration ranging from 0 to 20 mM. PMID- 15128102 TI - An amperometric inhibitor biosensor for the determination of reduced glutathione (GSH) without any derivatization in some plants. AB - In this study, an amperometric biosensor based on cucumber tissue homogenate was developed for the determination of glutathione. Cucumber (Cucumis sativus L.) tissue homogenate was used as the biological material. The cucumber tissue homogenate was cross-linked with gelatine using glutaraldehyde and fixed on a pretreated teflon membrane. The principle of the measurements was based on the determination of the decrease in the differentiation of oxygen level which had been caused by the inhibition of ascorbate oxidase in the biological material by glutathione. Determinations were carried out by standard curves which were obtained by the measurement of the decrease in the consumed oxygen level related to glutathione concentration. Optimization and characterization studies of the biosensor were carried out and a linearity in the gamma-L-glutamyl-L-cysteinyl glycine (GSH) concentration range 0.1-2 microM was obtained when 600 microM ascorbic acid was used as a substrate. The repeatability experiments (n = 7) revealed that for 1.5 microM GSH, the average value (x), standard deviation (S.D.) and variation coefficient (C.V.) were 1.517 microM, 4.72 x 10(-5) 3.11%, respectively. The biosensor useful lifetime was at least 2 months. The results of some plant samples analyzed with the presented biosensor agreed well with the spectrophotometric method (Ellman's reagent) used as a reference. PMID- 15128103 TI - Bio-photovoltaic conversion device using chlorine-e6 derived from chlorophyll from Spirulina adsorbed on a nanocrystalline TiO2 film electrode. AB - A bio-photovoltaic conversion device based on dye-sensitised solar cell (DSSC) using the visible light sensitisation of chlorine-e6 (Chl-e6) derived from chlorophyll from Spirulina adsorbed on a nanocrystalline TiO2 film was developed. Form fluorescence spectrum of Chl-e6 adsorbed on a nanocrystalline TiO2 film, the emission of Chl-e6 was effectively quenched by TiO2 nanocrystalline indicating that the effective electron injection from the excited singlet state of Chl-e6 into the conduction band of TiO2 particles occurred. The short-circuit photocurrent density (Isc). the open-circuit photovoltage (Voc). and the fill factor (FF) of solar cell using Chl-e6 adsorbed on a nanocrystalline TiO2 film electrode were estimated to be 0.305 +/- 0.012 mA cm(-2), 426 +/- 10 mV, and 45.0%, respectively. IPCE values were reached a maximum around the wavelength of absorption maximum (7.40% at 400 nm; 1.44% at 514 nm and 2.91% at 670 nm), indicating that the DSSC using visible light sensitisation of nanocrystalline TiO2 film by Chl-e6 was developed. PMID- 15128104 TI - Low-level detection of a bacillus anthracis simulant using Love-wave biosensors on 36 degrees YX LiTaO3. AB - We present an acoustic Love-wave biosensor for detection of the Bacillus anthracis simulant, Bacillus thuringiensis at or below inhalational infectious levels. The present work is an experimental study of 36 degrees YX cut LiTaO3 based Love-wave devices for detection of pathogenic spores in aqueous conditions. Given that the detection limit (D1) of Love-wave-based sensors is a strong function of the overlying waveguide, two waveguide materials have been investigated, which are polyimide and polystyrene. To determine the mass sensitivity of Love-wave sensor, bovine serum albumin (BSA) protein was injected into the Love-wave test cell while recording the magnitude and phase shift across each sensor. Polyimide had the lowest mass detection limit with an estimated value of 1.0-2.0 ng/cm2, as compared to polystyrene where D1 = 2.0 ng/cm2. Suitable chemistries were used to orient antibodies on the Love-wave sensor using protein G. The thickness of each biofilm was measured using ellipsometry from which the surface concentrations were calculated. The monoclonal antibody BD8 with a high degree of selectivity for anthrax spores was used to capture the non pathogenic simulant B. thuringiensis B8 spores. Bacillus subtilis spores were used as a negative control to determine whether significant non-specific binding would occur. Spore aliquots were prepared using an optical counting method, which permitted removal of background particles for consistent sample preparation. This work demonstrates that Love-wave biosensors are promising for low-level detection for whole-cell biological pathogens. PMID- 15128105 TI - Direct electron transfer and enzymatic activity of hemoglobin in a hexagonal mesoporous silica matrix. AB - The direct electrochemistry of hemoglobin (Hb) immobilized on a hexagonal mesoporous silica (HMS)-modified glassy carbon electrode was described. The interaction between Hb and the HMS was investigated using UV-Vis spectroscopy, FT IR, and electrochemical methods. The direct electron transfer of the immobilized Hb exhibited two couples of redox peaks with the formal potentials of -0.037 and 0.232 V in 0.1 M (pH 7.0) PBS, respectively, which corresponded to its two immobilized states. The electrode reactions showed a surface-controlled process with a single proton transfer at the scan rate range from 20 to 200 mV/s. The immobilized Hb retained its biological activity well and displayed an excellent response to the reduction of both hydrogen peroxide (H2O2) and nitrate (NO2-). Its apparent Michaelis-Menten constants for H2O2 and NO2- were 12.3 and 49.3 microM, respectively, showing a good affinity. Based on the immobilization of Hb on the HMS and its direct electrochemistry, two novel biosensors for H2O2 and NO2 were presented. Under optimal conditions, the sensors could be used for the determination of H2O2 ranging from 0.4 to 6.0 microM and NO2- ranging from 0.2 to 3.8 microM. The detection limits were 1.86 x 10(-9) M and 6.11 x 10(-7) M at 3sigma, respectively. HMS provided a good matrix for protein immobilization and biosensor preparation. PMID- 15128106 TI - Photoelectric response of polarization sensitive bacteriorhodopsin films. AB - Polarization sensitivity is introduced into oriented bacteriorhodopsin (BR) films through a photochemical bleaching process, which chemically modifies the structure of the purple membrane by breaking the intrinsic symmetry of the membrane-bound BR trimers. The resulting photovoltage generated in an indium-tin oxide (ITO)/BR/ITO detector is found to be anisotropic with respect to cross polarized probe beams. A model, based on the polarization dependent photoselection of the BR molecules qualitatively explains the photochemical bleaching process and the observed anisotropic response. The effect reported here can be used to construct a polarization sensitive BR-based bio-photoreceiver. PMID- 15128107 TI - Gold nanoparticle-based detection of genomic DNA targets on microarrays using a novel optical detection system. AB - The development of a nanoparticle-based detection methodology for sensitive and specific DNA-based diagnostic applications is described. The technology utilizes gold nanoparticles derivatized with thiol modified oligonucleotides that are designed to bind complementary DNA targets. A glass surface with arrays of immobilized oligonucleotide capture sequences is used to capture DNA targets, which are then detected via hybridization to the gold nanoparticle probes. Amplification with silver allows for detection and quantitation by measuring evanescent wave induced light scatter with low-cost optical detection systems. Compared to Cy3-based fluorescence, silver amplified gold nanoparticle probes provide for a approximately 1000-fold increase in sensitivity. Furthermore, direct detection of non-amplified genomic DNA from infectious agents is afforded through increased specificity and even identification of single nucleotide polymorphisms (SNP) in human genomic DNA appears feasible. PMID- 15128108 TI - Monolithically integrated bacteriorhodopsin/semiconductor opto-electronic integrated circuit for a bio-photoreceiver. AB - The light-sensitive protein, bacteriorhodopsin (BR), is monolithically integrated with an InP-based amplifier circuit to realize a novel opto-electronic integrated circuit (OEIC) which performs as a high-speed photoreceiver. The circuit is realized by epitaxial growth of the field-effect transistors, currently used semiconductor device and circuit fabrication techniques, and selective area BR electro-deposition. The integrated photoreceiver has a responsivity of 175 V/W and linear photoresponse, with a dynamic range of 16 dB, with 594 nm photoexcitation. The dynamics of the photochemical cycle of BR has also been modeled and a proposed equivalent circuit simulates the measured BR photoresponse with good agreement. PMID- 15128109 TI - Quantitative spectroscopy analysis of prokaryotic cells: vegetative cells and spores. AB - Multiwavelength ultraviolet/visible (UV-Vis) spectra of microorganisms and cell suspensions contain quantitative information on properties such as number, size, shape, chemical composition, and internal structure of the suspended particles. These properties are essential for the identification and classification of microorganisms and cells. The complexity of microorganisms in terms of their chemical composition and internal structure make the interpretation of their spectral signature a difficult task. In this paper, a model is proposed for the quantitative interpretation of spectral patterns resulting from transmission measurements of prokaryotic microorganism suspensions. It is also demonstrated that different organisms give rise to spectral differences that may be used for their identification and classification. The proposed interpretation model is based on light scattering theory, spectral deconvolution techniques, and on the approximation of the frequency dependent optical properties of the basic constituents of living organisms. The quantitative deconvolution in terms of the interpretation model yields critical information necessary for the detection and identification of microorganisms, such as size, dry mass, dipicolinic acid concentration, nucleotide concentration, and an average representation of the internal scattering elements of the organisms. E. coli, P. agglomerans, B. subtilis spores, and vegetative cells and spores of Bacillus globigii are used as case studies. It is concluded that spectroscopy techniques coupled with effective interpretation models are applicable to a wide range of cell types found in diverse environments. PMID- 15128110 TI - In vitro long-term performance study of a near-infrared fluorescence affinity sensor for glucose monitoring. AB - The long-term in vitro performance of a fluorescence affinity sensor for transdermal blood glucose monitoring was investigated. Affinity binding of fluorescently labeled concanavalin A (ConA) was used in this application, as previously described by Ballerstadt and Schultz [Anal. Chem. 17 (2000) 4185 4192). In this paper, the fluorescence emission of the sensor was extended to the near infrared (670 nm) using Alexa647 as the fluorochrome conjugated to concanavalin A. Sensors were alternately exposed to glucose solutions having concentrations of 2.5 and 20 mM with a dwell time of 3 h. The optical output of the sensors was monitored over a 4-month period. The sensors showed an initial increase in fluorescence over the first 3-4 weeks before gradually decreasing, with an approximately linear drop of 25% per month. In order to understand the reasons for the decrease in fluorescence output, further experiments were conducted, including time-dependent membrane leakage tests, solubility tests of ConA, temperature-dependent activity tests of ConA, and fluorescence photo bleaching tests. From these results, it became evident that the decrease in fluorescence was not due to denaturation of the ConA. The most likely cause was leakage of the fluorescently labeled ConA through the interface between the outer sealant and the membrane. This problem is considered to be solvable and future publications will address this issue. Extrapolation of the experimental data suggests that a leak-proof sensor would be remarkably stable with a fluorescence decrease of only 15% over a 1-year period. PMID- 15128111 TI - Modelling amperometric enzyme electrode with substrate cyclic conversion. AB - A mathematical model of amperometric enzyme electrodes in which chemical amplification by cyclic substrate conversion takes place in a single enzyme membrane has been developed. The model is based on non-stationary diffusion equations containing a non-linear term related to Michaelis-Menten kinetic of the enzymatic reaction. The digital simulation was carried out using the finite difference technique. The influence of the substrate concentration, the maximal enzymatic rate as well as the membrane thickness on the biosensor response was investigated. The numerical experiments demonstrate significant (up to dozens of times) gain in biosensor sensitivity at low concentrations of substrate when the biosensor response is under diffusion control. PMID- 15128112 TI - Neuronal differentiation and synapse formation of PC12 and embryonic stem cells on interdigitated microelectrode arrays: contact structures for neuron-to electrode signal transmission (NEST). AB - The development of neuron-microelectrode interfaces (neurochips) is highly desirable for the non-invasive recording of the cellular response to neuroactive drugs as well as the electrical stimulation of nervous tissue by implantable electrodes. A prerequisite for neuron-to-electrode signal transmission (NEST) is the formation of synapse-like contacts between the neuronal cell and the conductive surface of a microelectrode array. We attempted synapse formation by neuronal differentiation of rat pheochromocytoma cells (PC12) and blastocyst derived murine embryonic stem cells (ES-J1) on interdigitated microelectrode arrays that were made of gold (Au), platinum (Pt), or indium tin oxide (ITO). PC12 or ES cells were in vitro differentiated by incubation with nerve growth factor (NGF) and forskolin, or by serum deprivation and treatment with basic fibroblast growth factor (FGF-2), respectively. On top of ITO electrodes, the neuronal cells extended extremely long processes that terminated in pili-like contact structures, which is typical for growth cone formation. ES cells differentiated into neurons as verified by immunofluorescence staining of MAP-2 and developed synapse-like junctions with the ITO electrode surface as indicated by synaptophysin staining. Differentiated PC12 and ES cells showed bona fide morphological characteristics of synaptic growth cones that were unprecedented in tissue culture. Cones formed by PC12 cells could be stimulated with KCI and carbachol as shown by uptake of FM1-43, a fluorescent marker for synaptic vesicle formation. In contrast to Electrical Cell Impedance Spectroscopy (ECIS) recordings, AC impedance spectrometry with differentiated PC12 cells settled on interdigitated microelectrode arrays revealed lower AC impedance than that with undifferentiated cells, indicating that the complex impedance is dependent on ion fluxes at the neuron-to-electrode contact surface. PMID- 15128113 TI - Binding and dissociation kinetics using fractals: an analysis of electrostatic effects and randomly coupled and oriented coupled receptors on biosensor surfaces. AB - A fractal analysis is used to analyze the influence of: (a) electrostatic interactions on binding and dissociation rate coefficients for antibodies HH8, HH10, and HH26 in solution to hen egg-white lysozyme (HEL) immobilized on a sensor chip surface [Biophys. J. 83 (2002) 2946]; and (b) the binding and dissociation of recombinant Fab in solution to random NHS-coupled Cys-HEL and oriented thiol-coupled Cys-HEL immobilized on a sensor chip surface [Methods 20 (2000) 310]. Single- and dual-fractal models were employed to fit the data. Values of the binding and the dissociation rate coefficient(s) and the fractal dimensions were obtained from a regression analysis provided by Corel Quattro Pro 8.0 (Corel Corporation Limited, Ottawa, Canada. 1997). The binding rate coefficients are quite sensitive to the degree of heterogeneity on the sensor chip surface. It is of interest to compare the results obtained by the fractal analysis with that of the original analysis [Biophys. J. 83 (2002) 2946]. For example, as one goes from the binding of 21 nM HH10/HEL to the binding of 640 nM HH10/HEL(K97A), Sinha et al. [Biophys. J. 83 (2002) 29461 indicate that the enhancement of diffusional encounter rates may be due to 'electrostatic steering' (a long-range interaction). Our analysis indicates that there is an increase in the value of the fractal dimension, Df1 by a factor of 1.12 from a value of 2.133 2.385. This increase in the degree of heterogeneity on the surface leads to an increase in the binding rate coefficient, k1 by a factor of 1.59 from 12.92 to 20.57. The fractal analysis of binding and dissociation of recombinant Fab in solution to random NHS-coupled Cys-HEL and oriented thiol-coupled Cys-HEL immobilized on a sensor chip [Methods 20 (2000) 310] surface are consistent with the degree of heterogeneity present on the sensor chip surface for the random and the oriented case. As expected, the random case will exhibit a higher degree of heterogeneity than the oriented case, leading to subsequently a higher binding rate coefficient. PMID- 15128114 TI - Biomolecular reactions studied using changes in Brownian rotation dynamics of magnetic particles. AB - We have used magnetic particles to study specific binding of prostate specific antigen (PSA) to the surfaces of the bioparticles. The used particles have a mean diameter of about 130 nm and are placed in phosphate buffer saline (PBS). Each particle is composed of clusters of magnetic single domains of magnetite, which are covered with dextran. Changes in surface chemistry of the particles give rise to a change in the hydrodynamic volume of the particles. The later is mirrored by the changed frequency response of the complex magnetic susceptibility of a fluid containing these particles. Using ordinary induction coils and the lock-in amplifier technique it is possible to measure the complex magnetic susceptibility of the particle solution in a frequency range from about 10 Hz up to 10 kHz. From the measurement of the complex susceptibility versus the excitation frequency (both at the excitation frequency as well as at higher harmonics) we have shown that it is possible to quantitatively study the binding of PSA to the surfaces of the magnetic particles and thus to determine the PSA concentration in solution containing known concentration of nanoparticles functionalised with a monoclonal PSA antibody. Our method allows to perform an immunoassay in a single step and is much faster and cheaper compared to conventional ELISA procedures. PMID- 15128115 TI - Cerebral cortical oxygenation: a pilot study. AB - BACKGROUND: Cerebral hypoxia (cerebral cortical oxygenation [Pbro2] < 20 mm Hg) monitored by direct measurement has been shown in animal and small clinical studies to be associated with poor outcome. We present our preliminary results observing Pbro2 in patients with traumatic brain injury (TBI). METHODS: A prospective observational cohort study was performed. Institutional review board approval was obtained. All patients with TBI who required measurement of intracranial pressure (ICP), cerebral perfusion pressure (CPP), and Pbro2 because of a Glasgow Coma Scale score < 8 were enrolled. Data sets (ICP, CPP, Pbro2, positive end-expiratory pressure (PEEP), Pao2, and Paco2) were recorded during routine manipulation. Episodes of cerebral hypoxia were compared with episodes without. Results are displayed as mean +/- SEM; t test, chi2, and Fisher's exact test were used to answer questions of interest. RESULTS: One hundred eighty-one data sets were abstracted from 20 patients. Thirty-five episodes of regional cerebral hypoxia were identified in 14 patients. Compared with episodes of acceptable cerebral oxygenation, episodes of cerebral hypoxia were noted to be associated with a significantly lower mean Pao2 (144 +/- 14 vs. 165 +/- 8; p < 0.01) and higher mean PEEP (8.8 +/- 0.7 vs. 7.1 +/- 0.3; p < 0.01). Mean ICP and CPP measurements were similar between groups. In a univariate analysis, cerebral hypoxic episodes were associated with Pao2 < or = 100 mm Hg (p < 0.01) and PEEP > 5 cm H2O (p < 0.01), but not ICP > 20 mm Hg, CPP < or = 65 mm Hg, or Pac2 < or = 35 mm Hg. CONCLUSION: Cerebral oxymetry is confirmed safe in the patient with multiple injuries with TBI. Occult cerebral hypoxia is present in the traumatic brain injured patient despite normal traditional measurements of cerebral perfusion. Further research is necessary to determine whether management protocols aimed at the prevention of cerebral cortical hypoxia will affect outcome. PMID- 15128116 TI - Routinely repeated computed tomography after blunt head trauma: does it benefit patients? AB - BACKGROUND: Computed tomography of the head (HCT) is an integral part of the diagnosis and management of the patient with head injury, but the utility of repeated HCT performed solely for routine follow-up in the patient with blunt head trauma has not been defined. In the absence of clinical indications, routinely repeated HCT, even in patients with significant brain injury, does not contribute to patient care. METHODS: Trauma registry records at a Level I trauma center from July 1, 1997, to June 30, 2002, were reviewed. Patients with severe blunt head injury (Abbreviated Injury Scale score > or = 3) admitted to the intensive care unit and who had a repeat HCT scan obtained for scheduled follow up were included. Those patients with initial craniotomy, repeat HCT more than 72 hours after the initial HCT, or repeat HCT ordered for clinical indications were excluded. Data included were age, mechanism of injury, time to initial (HCT1) and repeat HCT (HCT2), indications for HCT2, and HCT findings. Additional data included Glasgow Coma Scale (GCS) score (admission and at HCT2); Injury Severity Score; occurrence of hypotension, coagulopathy, or elevated intracranial pressure (ICP); interventions made; and patient outcome. RESULTS: Entry criteria were met in 462 patients. Most were injured in motor vehicle crashes; the average age was 36 years and the mean initial GCS score was 9. The mean time to HCT1 was 1.3 hours and the mean time to HCT2 was 22.6 hours. HCT2 showed worsening in 85 patients (18.4%), and 16 patients had interventions in response to HCT2 (repeat HCT in 8, ICP monitoring or drainage in 6, and craniotomy in 2). No patient undergoing routine repeat HCT without other clinical findings required intervention. All patients with worsening HCT findings requiring intervention had coagulopathy, hypotension, ICP elevation, or marked decrease in GCS score. CONCLUSION: In the absence of clinical indicators or risk factors, repeat HCT after blunt head injury does not alter patient management and is unnecessary. PMID- 15128117 TI - Cerebral hypoxia in severely brain-injured patients is associated with admission Glasgow Coma Scale score, computed tomographic severity, cerebral perfusion pressure, and survival. AB - BACKGROUND: The purpose of this study was to determine the relationship of cerebral hypoxia with admission Glasgow Coma Scale (GCS) score, brain computed tomographic (CT) severity, cerebral perfusion pressure (CPP), and survival in patients with severe brain injury. METHODS: CPP and noninvasive transcranial oximetry (Stco2) were recorded hourly for 6 days in patients with a GCS score < or = 8 (3,722 observations). CT score was derived from midline shift (0/1) plus abnormal cisterns (0/1) plus subarachnoid hemorrhage (SAH) (0/1) (range, 0-3). RESULTS: Brain CT results were as follows: shift, 10 (56%); abnormal cisterns, 14 (78%); SAH, 9 (50%); epidural hematoma, 2 (11%); subdural hematoma, 11 (61%); and contusion, 17 (94%). The incidences of Stco2 < 60 were: GCS score 3-4, 26.5%; GCS score 5-7, 12.4%; and GCS score 8, 2.8% (p < 0.0001); CT score 2/3, 26.4%; and CT score 0/1, 10.0% (p < 0.0001); nonsurvivors 36.1%; and survivors 16.3% (p < 0.0001). For incidence of CPP < 70, the results were as follows: Stco2 < 60%, 33% of observations; Stco2 > or = 60%, 10% of observations (odds ratio, 4.3; p < 0.01). Despite CPP > or = 70, Stco2 < 60 incidence was 16% of observations. CONCLUSION: Cerebral hypoxia is common, even with CPP > or = 70, and is associated with GCS score, CT scan severity, and mortality. Cerebral hypoxia is related to cerebral hypoperfusion. Additional studies may prove that Stco2 monitoring will enhance the treatment of severe brain injury. PMID- 15128118 TI - Management of brain-injured patients by an evidence-based medicine protocol improves outcomes and decreases hospital charges. AB - OBJECTIVE: Traumatic brain injury (TBI) is the leading cause of death from blunt trauma, with an estimated cost to society of over dollar 40 billion annually. Evidence-based guidelines for TBI care have been widely discussed, but in hospital treatment of these patients has been highly variable. The purpose of this study was to determine whether management of TBI patients according to a protocol based on the Brain Trauma Foundation (BTF) guidelines would reduce mortality, length of stay, charges, and disability. METHODS: In 1995, a protocol following the BTF guidelines was developed by members of the Level I trauma center's interdisciplinary neurotrauma task force. Inclusion criteria for the protocol were blunt head injury, age > 14 years, and Glasgow Coma Scale score < or = 8. An extensive educational process was conducted to develop compliance among all disciplines for this new management strategy. A historical control group of patients eligible for the protocol was identified by retrospective analysis of trauma registry data for 1991 to 1994. Mortality, intensive care unit days, total hospital days, total charges, Rancho Los Amigos Scores, and Glasgow Outcome Scale scores were compared. RESULTS: Between 1991 and 2000, over 7,000 blunt TBI patients were managed by the Trauma Service. Of these, 830 met the inclusion criteria for the TBI protocol and lived > 48 hours. After implementation, initial analysis of the 1995-96 cohort indicated only 50% compliance with the protocol. By 1997, compliance had risen to 88%. Patients were therefore compared as three groups: before the protocol (1991-94, n = 219), during low compliance (1995-96, n = 188), and during high compliance (1997-2000, n = 423). Groups did not differ significantly on Injury Severity Score, head Abbreviated Injury Scale score, or age (p > 0.05). Admission Glasgow Coma Scale score was slightly higher in the 1991-94 cohort (4.0 vs. 3.5, p = 0.001). From 1991-94 to 1997-2000, intensive care unit stay was reduced by 1.8 days (p = 0.021) and total hospital stay was reduced by 5.4 days (p < 0.001). The charge reduction (calculated in 1997 dollars) per patient for the length of stay decrease was dollar 6,577 in 1995-96 and dollar 8,266 in 1997-2000 (p = 0.002). This represents a total reduction over 6 years of dollar 4.7 million in charges. In addition, the overall mortality rate showed a reduction of 4.0% from 1991-94 to 1997-2000 (17.8% vs. 13.8%), although this was not statistically significant. On the basis of the Glasgow Outcome Scale score, in 1997-2000, 61.5% of the patients had either a "good recovery" or only "moderate disability," compared with 503% in 1995-96 and 43.3% in 1991-94 (p < 0.001). The Rancho Los Amigos Scores showed a similar trend, with 56.6% of the 1997-2000 patients having appropriate responses at 10 to 14 days, compared with only 44.0% of the 1995-96 patients and 43.9% of the 1991-94 patients (p = 0.004). CONCLUSION: Adherence to a protocol based on the BTF guidelines can result in a significant decrease in hospital days and charges for TBI patients who live > 48 hours. In addition, mortality and outcome may be significantly affected. This analysis suggests that increased efforts to improve adherence to national guidelines may have a significant impact on head injury care outcomes and could dramatically reduce the substantial financial resources that are currently consumed in the acute care phases for this injury. PMID- 15128120 TI - The influence of side airbags on the risk of head and thoracic injury after motor vehicle collisions. AB - BACKGROUND: Side airbags (SABs) were developed as an energy-absorbing barrier to protect specific occupant body regions in near side impact motor vehicle collisions. METHODS: The National Automotive Sampling System Crashworthiness Data System was used to evaluate drivers and front-seated passengers in 1998 or newer vehicles involved in near side impact collisions. Risk ratios were calculated comparing the risk of head and thoracic injury among occupants in vehicles with and without SABs adjusting for occupant, vehicle, and collision characteristics. RESULTS: Occupants in vehicles equipped with head protection SABs had a 75% lower risk of head injury (p = 0.008) after near side collisions. With respect to thoracic injury, SABs that provide thoracic protection are associated with a 68% reduction (p = 0.01) in thoracic injury risk. CONCLUSION: As SAB-equipped vehicles become an increasingly larger segment of the on-road vehicle fleet, the impact of head and thoracic injury after near side impact collision is likely to be reduced. PMID- 15128119 TI - Inhibition of enteral enzymes by enteroclysis with nafamostat mesilate reduces neutrophil activation and transfusion requirements after hemorrhagic shock. AB - BACKGROUND: The gut origin of the inflammatory response in trauma patients has been difficult to define. "In vivo" generation of neutrophil-activating factors by gut proteases may be a cause of multiorgan failure after hemorrhagic shock, and can be prevented with the serine protease inhibitor nafamostat mesilate (Futhan). The objective of this study was to determine the effect of nafamostat mesilate given by enteroclysis on enteric serine protease activity, neutrophil activation, and transfusion requirements during hemorrhagic shock. METHODS: Sixteen pigs weighing 21 to 26 kg were divided into control and treatment groups. A laparotomy was performed under anesthesia, and catheters were placed in the duodenum, midjejunum, and terminal ileum. Pigs were bled 30 mL/kg over 30 minutes and maintained at a mean arterial pressure of 30 mm Hg for 60 minutes. Shed blood was then used to maintain a mean arterial pressure of 45 mm Hg for another 3 hours. Treated animals received 100 mL/kg of 0.37 mmol/L nafamostat mesilate in GoLYTELY through the duodenal catheter at 1 L/h. Control animals received GoLYTELY only. Samples of enteral content and blood were taken at baseline, after shock, and at 30-minute intervals during resuscitation. Animals were killed after 3 hours of resuscitation. Enteral trypsin-like activity at the three gut sites was measured by spectrophotometry. Activation of naive human neutrophils by pig plasma was measured by the percentage of cells having pseudopods larger than 1 microm on microscopy. Lung, liver, and small bowel were analyzed by histology and myeloperoxidase assay. RESULTS: Both control and nafamostat mesilate-treated groups had significant reductions in protein and protease levels in the duodenum during enteroclysis; however, only nafamostat mesilate-treated animals had persistent suppression of protease activity throughout the experiment. Nafamostat mesilate-treated animals had a lower transfusion requirement of shed blood, 18.1 +/- 4.5 mL/kg versus 30 +/- 0.43 mL/kg (p = 0.002). Nafamostat mesilate-treated animals had significantly less neutrophil activation than controls at 150 minutes after resuscitation (33.7 +/- 6.48% vs. 42.4 +/- 4.57%,p = 0.01) and 180 minutes after resuscitation (31.1 +/- 3.31% vs. 46.9 +/- 4.53%, p = 0.0002). Lung myeloperoxidase activity was lower in nafamostat mesilate-treated animals (0.31 +/- 0.14) than in control animals (0.16 +/- 0.04, p = 0.04). Histology of liver and small intestine showed less injury in nafamostat mesilate-treated animals. CONCLUSION: Nafamostat mesilate given by means of enteroclysis with GoLYTELY significantly reduces enteral protease levels, leukocyte activation, and transfusion requirements during resuscitation from hemorrhagic shock. This strategy may have clinical promise. PMID- 15128121 TI - Can we rely on computed tomographic scanning to diagnose pulmonary embolism in critically ill surgical patients? AB - BACKGROUND: Spiral computed tomographic pulmonary angiography (CTPA) is gaining an increasing role in pulmonary embolism (PE) diagnosis because it is more convenient and less invasive than conventional pulmonary angiography (PA). Encouraging reports on the reliability of CTPA for medical patients have prompted widespread use despite the fact that its value in critically ill surgical patients has been inadequately explored. Hemodynamic and respiratory issues of critical illness may interfere with CTPA's diagnostic accuracy. The objective of this study was to compare CTPA with PA for the diagnosis of PE in critically ill surgical patients. METHODS: Over 30 months (August 1999-February 2002), 37 critically ill surgical patients (28 trauma and 9 non-trauma patients) wiith clinical suspicion of PE were enrolled prospectively. CTPA and PA were independently interpreted by four radiologists (two for each test) blinded to each other's interpretation. Clinical suspicion for PE was classified as high, intermediate,or low on the basis of predetermined criteria. PA was considered as the standard of reference for the diagnosis of PE. RESULTS: PE was found in 15 (40%) patients by: central PE in 8 and peripheral PE in 7. CTPA and PA findings were different in 11 patients (30%): CTPA was false-negative in 9 patients and false-positive in 2. Its sensitivity and specificity were PE 50% and 100%, respectively, for central PE; 28% and 93% for peripheral PE; and 40% and 91% for all PE. There were no differences in risk factors or clinical characteristics between patients with and without PE. The level of clinical suspicion was identical in the two groups. The independent reviewers disagreed on CTPA or PA interpretations in 11% and 16% of the readings, respectively. CONCLUSION: PA remains the "gold standard" for diagnosis of PE in critically ill surgical patients. CTPA should be explored further before being universally accepted. Clinical criteria are unreliable for detecting PE in this population and therefore a high index of suspicion should be maintained. PMID- 15128122 TI - Surgeon-performed ultrasound for pneumothorax in the trauma suite. AB - BACKGROUND: Surgeon-performed ultrasound has become ubiquitous in the trauma suite. Initial reports suggest that sonography may be used for the detection of pneumothorax. The purpose of this study was to evaluate the efficacy of sonography to rule out the presence of a pneumothorax in the trauma population. METHODS: A prospective analysis of 328 consecutive trauma patients at an American College of Surgeons-verified Level I trauma center was undertaken. Thoracic ultrasound was performed before chest radiography. The presence or absence of a "sliding-lung" sign or "comet-tail" artifact was recorded. RESULTS: Of 328 evaluations, there were 312 true-negatives, 12 true-positives, 1 false-negative, 1 false-positive, and 2 exclusions. Specificity, negative predictive value, and accuracy were 99.7%, 99.7%, and 99.4%, respectively. CONCLUSION: Ultrasound is a reliable modality for the diagnosis of pneumothorax in the injured patient. This modality may serve as an adjunct or precursor to routine chest radiography in the evaluation of injured patients. PMID- 15128123 TI - Prehospital endotracheal intubation for trauma does not improve survival over bag valve-mask ventilation. AB - BACKGROUND: Few data exist supporting a survival benefit to prehospital endotracheal intubation (ETI) over bag-valve-mask ventilation (BVM) in trauma patients. METHODS: Data were reviewed from all trauma patients transported to our Level I trauma center receiving prehospital ETI or BVM. Mortality was adjusted by age, Revised Trauma Score, Injury Severity Score, and mechanism of injury (penetrating vs. blunt). RESULTS: Of 5,773 patients, 316 (5.5%) had ETI and 217 (3.8%) had BVM. Patients receiving ETI were significantly more like to die (88.9% vs. 30.9%, p < 0.0001). When corrected for Injury Severity Score, Revised Trauma Score, and mechanism of injury, ETI was associated with similar or greater mortality than BVM. ETI patients had longer prehospital times (22.0 vs. 20.1 minutes, p = 0.0241). CONCLUSION: In our trauma system, when corrected for mechanism and severity of anatomic and physiologic injury, ETI confers no survival advantage over BVM and slightly increases prehospital time. PMID- 15128124 TI - Contrast extravasation predicts the need for operative intervention in children with blunt splenic trauma. AB - BACKGROUND: Although the presence of a contrast blush (CB) on computed tomographic (CT) scan is associated with an increased failure rate of nonoperative management in adults with blunt splenic injury, little information is available for the pediatric population, where nonoperative management is the standard of care. Our aim was to determine whether the finding of CB on CT scan could predict failure of nonoperative therapy in children with blunt splenic injury. METHODS: A retrospective analysis of 343 patients admitted with blunt splenic injury to our Level I pediatric trauma center over a 7-year period was performed. All CT scans were reviewed by a radiologist who was blinded to the patient outcome. We excluded 127 patients who either underwent immediate laparotomy without a CT scan or whose CT scans were unavailable at the time of this review. We divided the patients into two groups on the basis of the presence or absence of CB on the updated reading of the CT scan. Demographic variables analyzed included age, sex, mechanism of injury, Injury Severity Score, Glasgow Coma Scale score, initial hemoglobin and hematocrit, and emergency department pulse rate and systolic blood pressure. Outcome measures compared include length of stay, length of intensive care unit stay, the need for splenic intervention, and mortality. Continuous variables were compared using Student's t test for normally distributed data and the Mann-Whitney test for skewed data. Categorical data were compared using chi2 analysis or Fisher's exact test. Statistical significance was assigned to values of p < 0.05. RESULTS: Among the study population (N = 216), 27 patients (12.5%) had CB on CT scan. Patients with CB had significantly lower hematocrit (p = 0.0004) and required operative intervention more frequently than those without CB (22% vs. 4%;p = 0.0008). Among patients with CB, mean pulse rate at presentation was higher in those that required splenic intervention (SI) (129 +/- 20.1) compared with those who underwent successful nonoperative therapy (100.4 +/- 23.1; p = 0.01). Only grade V injuries correlated with the need for laparotomy. CONCLUSION: Children with blunt splenic injury who have CB on CT scan are more likely to require SI than those without CB. However, because the majority of patients with CB did not require SI, in the absence of hemodynamic instability, this finding may be insufficient to determine the need for SI. CB is a specific marker of active bleeding that may predict the need for early splenic intervention in a specific subset of patients at presentation. PMID- 15128125 TI - Splenic embolization revisited: a multicenter review. AB - BACKGROUND: Splenic embolization can increase nonoperative salvage. However, complications are not clearly defined. A retrospective multicenter review was performed to delineate the risks and benefits of splenic embolization. METHODS: A retrospective chart review of all patients undergoing splenic embolization from 1997 to 2002 at four separate Level I trauma centers was performed. Reviewed results included patient demographics, admission and follow-up computed tomographic scan results, angiographic technique, and patient outcomes including splenic salvage rate and procedural complications. RESULTS: A total of 140 patients were reviewed. The majority were young male patients involved in motor vehicle crashes. These patients had high abdominal computed tomographic grades of splenic injury and moderate Injury Severity Scores. The splenic salvage rate was 87%, which decreased with increasing injury grade. However, over 80% of splenic injury grades 4 and 5 were successfully managed nonoperatively. Significant hemoperitoneum did not affect success, but the presence of arteriovenous fistula was associated with a high failure rate, even with embolization. Salvage rates were similar between main coil and subselective embolization groups. Patients over 55 years of age did no worse than younger patients. Major complications included bleeding in 16 patients; 6 splenic abscesses, with 5 patients requiring splenectomy; and 1 episode of arterial injury requiring operative repair. CONCLUSION: Splenic embolization remains a valuable technique in splenic salvage, especially in higher grade injuries. Complications are common but do not seem to affect outcome. PMID- 15128126 TI - Heterogeneity in the effect of albumin and other resuscitation fluids on intracellular oxygen free radical production. AB - BACKGROUND: We hypothesized that by studying a suspension of a single cell type in various resuscitation fluids, we could compare their effects on parameters associated with cell death. METHODS: Jurkat cells were suspended in resuscitation fluids. Using flow cytometry, light scatter, phosphatidylserine translocation, propidium iodide uptake, and intracellular H2O2 production were measured. RESULTS: Buffer (pH, 7.4) and albumin added to Ringer's lactate inhibited the adverse changes, including intracellular oxygen free radical production. Oxygen free radical production was variable within the cell population and inhibited by albumin but not other colloids or crystalloids. This correlated well with the ability of albumin to enhance metabolic activity. A flavoprotein inhibitor blocked H2O2 production, suggesting that mitochondria are the source of the H2O2 and the variability. CONCLUSION: Oxygen free radical inhibition by albumin could explain both its beneficial and its harmful effects. PMID- 15128127 TI - Management of destructive bowel injury in the open abdomen. AB - BACKGROUND: Little attention has been focused on destructive injuries of the bowel in patients requiring open abdominal management. We therefore reviewed our institutional experience for destructive bowel injury requiring open abdominal management with the vacuum pack technique (vac). METHODS: The trauma registry at a Level I trauma center was used to identify patients sustaining destructive bowel injury for an 11-year period beginning in May 1990. Patients were assessed for pertinent clinical and demographic information, and individuals requiring open abdominal management were compared with those who did not. RESULTS: One hundred four patients required bowel resection and constitute the study population. Twenty-nine patients had vacs placed, with 22 (75.9%) of the total eventually obtaining delayed fascial closure. Nineteen (183%) patients had resection and primary repair (PR) of large and/or small bowel in conjunction with a vac, 10 (9.6%) patients had stoma formation in conjunction with a vac, 62 (59.6%) patients had resection and PR of small and/or large bowel in conjunction with primary fascial closure, and 13 (12.5%) patients had stoma formation and primary fascial closure. There were no differences in abdominal abscess or leak rates between groups. There were four deaths, none of which was secondary to failure of an anastomosis. CONCLUSION: Bowel resection with PR appears to be a safe alternative after destructive bowel injury and results in acceptable morbidity when performed in conjunction with open abdominal management. PMID- 15128129 TI - Alleviated tension at the repair site enhances functional regeneration: the effect of full range of motion mobilization on the regeneration of peripheral nerves--histologic, electrophysiologic, and functional results in a rat model. AB - BACKGROUND: In the clinical management of combined tendon and nerve injuries, competing treatment strategies are well known. The effect of mobilization on the functional regeneration of peripheral nerves remains controversial. This study sought to determine the effect of full range of motion mobilization on nerve repair by using tubular segmental nerve splinting to block movement, and thereby variable tension, at the nerve repair site. METHODS: In 96 rats, the right sciatic nerve was transected midthigh and coapted immediately microsurgically. The groups used in the study were as follows: group N, epineural nerve repair; group T, segmental tubular nerve splinting with fixed in situ tension at the nerve suture site,allowing segmental movement only; group TN, segmental tubular nerve splinting with alleviated in situ tension at the nerve suture site, allowing segmental movement only; and group TM, segmental tubular nerve splinting without fixed in situ tension at the nerve suture site, allowing movement of the nerve suture site. Full range of motion of the lower limbs was ensured by passive motion of hind limbs once a week after functional testing. Blinded histologic, immunohistochemical, and electrophysiologic assessment and 12 postoperative weekly function tests were carried out. RESULTS: Functional and electrophysiologic results were significantly better in group TN, by segmental tubular nerve splinting with alleviated in situ tension at the nerve repair site, mainly because of less scar formation and enhanced endoneural angiogenesis at the nerve suture segment. CONCLUSION: Full range of motion mobilization may impede functional nerve recovery by significant endoneural collagenization and decreased angiogenesis at the nerve suture segment. Complete alleviation of in situ (pathophysiologic) tension at the nerve suture site seems to improve functional peripheral nerve regeneration. PMID- 15128128 TI - Management of blunt thoracic aortic injuries: endovascular stents versus open repair. AB - BACKGROUND: Endovascular stent graft (EV) technology has been successfully adapted to the repair of blunt traumatic aortic injuries. The purpose of this study was to compare the outcomes of patients treated with EV repair and open repair after blunt thoracic aortic trauma. METHODS: A review of a tertiary trauma center's prospective trauma registry identified all patients who suffered a blunt traumatic thoracic aortic injury over an 11-year period (1991-2002). Operative interventions and outcomes were then compared. RESULTS: Over an 11-year period, 18 patients underwent repair of a blunt thoracic aortic injury (EV, 6; open, 12). There were no significant differences in demographics, injury, or crash statistics between groups. The open group had a 17% early mortality rate (n = 2), a paraplegia rate of 16% (n = 2), and an 8.3% incidence of recurrent laryngeal nerve injury (n = 1). This is in contrast to a 0% rate of mortality, paraplegia, and recurrent laryngeal nerve injury in the EV group. A definite trend toward decreased morbidity, mortality, intensive care unit length of stay, and number of ventilator-dependent days was seen with EV repair. CONCLUSION: We observed a clear trend toward improved outcomes after EV repair of thoracic aortic injuries compared with standard open repair. EV repair is emerging as the preferred method of repairing blunt thoracic aortic injuries in trauma patients with multiple injuries. PMID- 15128130 TI - Effect of gamma-hydroxybutyric acid on lipid peroxidation and tissue lactate level in experimental head trauma. AB - BACKGROUND: This study was designed to determine the effects of gamma hydroxybutyric acid (GHB) on tissue lactate and malondialdehyde (MDA) levels in rabbit brain after experimental head trauma. METHODS: Thirty New Zealand rabbits were divided equally into three groups: group S was the sham-operated group, group C, and group GHB received head trauma, where group C was the untreated and group GHB was the treated group. Head trauma was delivered by performing a craniectomy over the right hemisphere and dropping a weight of 10 g from a height of 80 cm. GHB was administered 400 mg/kg intravenously for 10 minutes after the head trauma to group GHB. The nontraumatized side was named "1" and the traumatized side was named "2." One hour after trauma, brain cortices were resected from both sides and the concentrations of lactate and MDA were determined. RESULTS: There were significant differences between lactate and MDA levels of group S and all other groups (C1, C2, GHB1, and GHB2) except between lactate levels of group S and group GHB1, the nontraumatized and traumatized sides of groups C and group GHB, group C2 versus group GHB2, and group C1 versus group GHB1 (p < 0.05). Rectal temperature after the administration of GHB in group GHB was found lower than in groups S and C (p < 0.05). CONCLUSION: These results demonstrate that head trauma leads to an increase in brain tissue lactate and MDA levels, and GHB effectively suppresses the increase of lactate and MDA. PMID- 15128131 TI - Regional heterogeneity of cerebral blood flow response to graded pressure controlled hemorrhage. AB - BACKGROUND: Little is known about the regional distribution of cerebral blood flow (CBF) in nonanesthetized animals during periods of lowered blood pressure. The present investigation addresses the specific reaction patterns of local cerebral blood flow (LCBF) in comparison with mean CBF during graded pressure controlled hemorrhagic shock in conscious rats. METHODS: Conscious rats were subjected to graded pressure-controlled hemorrhage (to 85, 70, 55, or 40 mm Hg) by arterial blood withdrawal. After a period of 30 minutes, blood pressure was stabilized by withdrawal or reinfusion of blood. LCBF was determined autoradiographically by the iodo(14C)antipyrine method in 34 brain structures, and mean CBF was calculated and compared with the values of nonhemorrhaged control animals. RESULTS: Mean CBF remained unchanged except for the group with the lowest blood pressure of 40 mm Hg (decrease in CBF of 28%). Otherwise, LCBF was increased in some brain structures at an unchanged mean CBF. Congruently, at 40 mm Hg, the decrease in mean CBF did not show up in all brain structures, the local pattern of CBF varying between an unchanged and a profoundly decreased CBF. The mean coefficient of variation of CBF was increased with the severity of hemorrhagic shock, which indicates an enhanced heterogeneity of CBF. CONCLUSION: Because of the substantial heterogeneity in the responses of LCBF to pressure controlled hemorrhage, autoregulation of CBF during pressure-controlled hemorrhagic shock has to be reconsidered on a regional basis. PMID- 15128132 TI - Alterations of myelin basic protein and ultrastructure in the limbic system at the early stage of trauma-related stress disorder in dogs. AB - BACKGROUND: The secondary injury and related complications after trauma are still the focus of trauma research. However, whether the remote effects on the central nervous system could be induced by high-energy missile extremity impact remains unclear. Also, the possible biomarker for brain damage in traumatic stress disorder has not been determined. METHODS: Forty-two healthy adult dogs were divided into three groups: the control group (n = 12), the high-speed trauma group (n = 15), and the low-speed trauma group (n = 15). Bilateral thighs of dogs were wounded with a smoothbore 6.2-mm rifle at a speed of 1,368 m/s (1.03-g steel bullet) for the high-speed trauma group and 625 m/s for the low-speed trauma group. The expression of myelin basic protein (MBP) in cerebrospinal fluid (CSF), hypothalamus and hippocampus of the limbic system, and temporoparietal cortex was investigated by enzyme-linked immunosorbent assay and dot-blot analysis. Also, the ultrastructure of the above areas was observed with light and electron microscopy. RESULTS: Neuronal degeneration and nerve fiber demyelination were seen in the hypothalamus and hippocampus in the high-speed trauma group at 8 hours after impact. The MBP level was markedly increased in the CSF (p < 0.01) in the two trauma groups, in the hypothalamus of the low-speed trauma group (p < 0.05), and in both the hypothalamus and the hippocampus of the high-speed trauma group (p < 0.01). The expression of MBP mRNA was also significantly enhanced in these areas at the same time. The increase of MBP content in the CSF was positively correlated with the elevation of MBP concentration in the hypothalamus and hippocampus. CONCLUSION: The hypothalamus and hippocampus of the limbic system in the central nervous system are vulnerable to damage after high-energy missile extremity impact, indicating that it might be one of the important pathologic bases involved in the development of trauma-related complications. Meanwhile, the MBP level in the CSF may be a sensitive biological indicator for brain damage at the early stage of trauma-related stress disorder. PMID- 15128133 TI - The role of oxygen-free radical in the apoptosis of enterocytes in scalded rats after delayed resuscitation. AB - BACKGROUND: This study aimed to evaluate the relation between apoptosis of enterocytes and oxygen-free radical injury in scalded rats with delayed resuscitation as well as the role of antioxidants in the prevention of enterocyte apoptosis. METHODS: For this study, 150 male Wistar rats were divided randomly into four groups representing early resuscitation (ER), delayed resuscitation (DR), N-acetylcysteine (NAC) treatment, and allopurinol (Allo) treatment. The animals were subjected to a 30% total body surface area, full-thickness scald. Fluid therapy was started 6 hours after the injury in the DR and treatment groups. Apoptosis of enterocytes was identified by DNA fragmentation (ap%), DNA agarose gel electrophoresis, and terminal deoxynucleotidyl transferace (TdT) mediated dUPT-biotin nick end labeling (TUNEL). The contents of malondialdehyde (MDA), total sulfhydryl (TSH), and nonprotein sulfhydryl (NPSH) and the activity of xanthine oxidase in intestinal mucosa were determined after the burn in the four groups. RESULTS: Apoptosis of enterocytes increased significantly in all the groups. The animals in the DR group showed an earlier and greater increase in ap% than the animals in the ER group. Similar results were seen for electrophoresis, TUNEL assay, and levels of MDA, xanthine oxidase (XO), TSH, and NPSH. Treatment with NAC was associated with a decrease in ap% and MDA, but not XO, as compared with the levels in the DR group, whereas treatment with Allo was associated with a decrease in MDA and XO, but not ap%. Delayed resuscitation was associated with significant decreases in TSH and NPSH, as compared with the levels in the ER group, whereas both the NAC and Allo groups had significantly higher levels of TSH and NPSH than the DR group. CONCLUSIONS: Significant apoptosis of enterocytes was induced by oxidative stress in the intestinal mucosa after a burn in rats. The findings show that NAC blunted intestinal apoptosis induced by oxygen-free radical, which was generated in the process of ischemia-reperfusion injury after a burn because of delayed resuscitation. PMID- 15128134 TI - Evaluation of renal cortical perfusion by noninvasive power Doppler ultrasound during vascular occlusion and reperfusion. AB - BACKGROUND: Urine output, a frequently used resuscitation end point, is presumed to represent renal cortical perfusion. However, no noninvasive method for direct measurement of renal perfusion exists. Power Doppler ultrasound (PDUS) is a method that reportedly is sensitive to low-velocity and microvascular blood flow and can depict it. This study aimed to develop a quantitative technique for PDUS image analysis, and to evaluate the ability of PDUS to quantify cortical perfusion during renal ischemia induced by vascular occlusion. METHODS: A method was developed to determine the mean gray-scale intensity of PDUS images from within the renal cortex (PDUS image intensity). This index was hypothesized to represent renal cortical microvascular blood flow. Renal cortical blood flow was determined using fluorescent microspheres in five swine. Renal artery flow was measured with an ultrasonic flow probe. Power Doppler ultrasound was performed at baseline; at 75%, 50%, and 25% of baseline renal artery flow; and during reperfusion. RESULTS: Subjectively, PDUS images showed decreases in image intensity corresponding to renal artery occlusion and increases after reperfusion. Cortical blood flow correlated well with renal artery flow (n = 25; r2 = 0.868) and with PDUS image intensity (n = 25; r2 = 0.844). CONCLUSION: Noninvasive power Doppler ultrasound image intensity correlated well with invasively measured renal cortical blood flow, and may be useful during resuscitation of injured and critically ill patients. PMID- 15128135 TI - Helical computed tomographic angiography for the diagnosis of traumatic arterial injuries of the extremities. AB - BACKGROUND: The radiologic study of choice for evaluation of traumatic arterial injuries is conventional arteriography, but it poses the risks of an invasive procedure. Computed tomographic arteriography (CTA) is emerging as a new way to study arterial anatomy, with the additional advantages of being noninvasive and a technique that allows evaluation of different body areas simultaneously. Our experience using CTA for evaluation of traumatic arterial injuries is provided in this study. METHODS: A retrospective review over a 22-month period of all adult patients undergoing CTA for evaluation of traumatic injuries to the extremities was performed. RESULTS: A total of 97 CTA studies were performed in the 95 patients. CTA adequately demonstrated the nature and location of all the arterial injuries when compared with conventional arteriography or surgical exploration. Abnormal CTA results included 21 arterial occlusions, 2 intimal flap defects, and 2 pseudoaneurysms. Nine of these 25 injuries were confirmed by surgery only, 10 by surgery and arteriography, and 6 by arteriography only. Normal CTA results were confirmed with arteriography in 10 cases. No missed injuries were encountered in patients with normal CTA results. CONCLUSION: CTA is a reliable technique for the detection and characterization of traumatic extremity arterial injuries. These results suggest CTA may be an alternative to conventional arteriography for the diagnosis of traumatic arterial injuries. PMID- 15128136 TI - Vertebroplasty for the treatment of osteoporotic compression spinal fracture: comparison of remedial action at different stages of injury. AB - BACKGROUND: Outstanding results have been achieved using vertebroplasty for the treatment of osteoporotic compression spinal fractures, and a number of studies have reported the results from the application of this procedure. This study compared the results of vertebroplasty used at the different stages of injury. METHODS: A retrospective study reviewing the period between January 2001 and July 2001 investigated 68 patients who underwent single-level vertebroplasty: 22 patients within 2 weeks of the injury, 22 patients 2 weeks to 2 months after the injury, and 24 patients more than 2 months after the injury. Clinical evaluations compared the results of treatment at different injury stages during a mean follow up period of 13 months. RESULTS: Although all the patients undergoing vertebroplasty in the acute and subacute stages reported satisfaction within 1 week of the operation, only 72.7% of the acute-stage group reported satisfaction with 24 hours of surgery. Moreover, evidence of cement leakage after vertebroplasty was detected for 27.3% of the acute-stage patients. This percentage significantly higher than for the patients in the subacute and chronic stages. Radiographic examination showed that new, adjacent compression fracture had occurred for 10.3% of the patients, with anterior interbody restabilization occurring for 11.8%. CONCLUSIONS: The results for vertebroplasty treatment of osteoporotic compression fractures appear to be injury stage dependent, with patients in the acute-injury stage needing longer recovery times, and with cement leakage quite common. These findings lead to the conclusion that the subacute stage is optimal for vertebroplasty. Furthermore, it is suggested that the use of spinal orthoses and postsurgical supplementation for the bone matrix reduces the risk of new, adjacent compression fractures and increases anterior interbody restabilization. Importantly, the findings suggest that a presurgical magnetic resonance imaging evaluation is an absolute necessity. PMID- 15128137 TI - Long-term results of multiple-stage treatment for posttraumatic osteomyelitis of the tibia. AB - BACKGROUND: The treatment of posttraumatic osteomyelitis of the tibia requires meticulous debridement and adequate soft tissue coverage. At our institution, we perform a staged procedure consisting of surgical debridement followed by muscle coverage. If necessary, implantation of a cancellous iliac bone graft was always performed as a three-stage treatment. METHODS: We performed a retrospective analysis of 47 patients treated for posttraumatic osteomyelitis of the tibia between 1987 and 1998. RESULTS: Twenty-two patients originally had a Gustilo grade III fracture, 21 patients had a Gustilo grade I or II or closed fracture, the Gustilo grade was not known for 2 patients, and 2 patients had no fracture. Using the Cierny-Mader classification, most patients had a localized osteomyelitis. To cover the debrided area, 20 pedicled muscle transfers and 28 microvascular free flaps were used; one patient had two localizations of osteomyelitis (both proximal and distal) and received two muscle flaps. Flap failure was 8% and was successfully treated by additional flap coverage in two cases; one was closed by a split skin graft and one was closed by secundum. Twenty-six patients received a cancellous bone graft. During an average follow-up of 94 months, 9% had a recurrence of osteomyelitis for which additional surgical interventions were necessary. Finally, all the infections were eventually cured. CONCLUSION: Our staged surgery proved to be an excellent method of treating osteomyelitis after open or closed fractures of the tibia. PMID- 15128138 TI - The mosaic of equestrian-related injuries in Greece. AB - BACKGROUND: This study aims to estimate the burden and describe the profile of equestrian injuries in Greece, where horses, donkeys, and mules are still used in agriculture and where horse riding is a popular leisure activity. METHODS: Prospectively collected information on 140,823 injuries reported in the national Emergency Department Injury Surveillance System was examined and 244 equestrian related injuries that occurred during farming, equestrian sports, or horse racing were analyzed. RESULTS: The estimated countrywide injury incidence for farming and equestrian sports combined was 21 per 100,000 person-years, but it was 160 times higher for horse-racing personnel. Men had higher rates of racing injuries and women had higher rates of equestrian sport injuries. Fractures accounted for 39.0% of injuries in horse racing and 30.5% in farming; head injuries accounted for approximately 50% of injuries among farmers. Farming injuries were more serious, with 25% requiring hospitalization. Analysis through the Barell matrix pointed to the role of spurs in the causation of ankle fractures and dislocations and the likely contribution of helmets in preventing traumatic brain injuries. CONCLUSION: Equestrian-related injuries are a serious but underappreciated health problem and merit targeted prevention efforts for each category affected. PMID- 15128139 TI - Visual outcome and ocular survival after sports related ocular trauma in playing golf. AB - PURPOSE: To study eye injuries. More specifically, to study the visual performance and ocular findings in people generating the eye trauma while golfing and to define the main mechanisms of injury. METHODS: We retrospectively reviewed a case series of seven patients with golf-related eye injuries treated at Austrian hospitals over the last seven years in a multicenter setting. RESULTS: Five men and two women received treatment. The mean age was 46 (range, 29-63), and three of the patients had blunt close globe trrauma and four suffered from a ruptured globe. Following the injuries, the patients' initial visual acuity ranged from no light perception to 20/40. All of the patients recquired surgery, with the resulting visual acuily ranging from moving hands to 20/20. It should be noted that three eyes required enucleation. CONCLUSION: Visual outcome from surgery is frequently very poor in golf related injuries. This is further complicated by these traumas frequently being accompanied by a high enucleation rate. Among the patients reviewed, it was observed that ruptured globe trauma has a worse prognosis than close blunt trauma. Due to the severe resulting complications of golf-related ocular injuries, we believe eye protection should be considered and emphasized. PMID- 15128140 TI - Analysis of injury criteria to assess chest and abdominal injury risks in blunt and ballistic impacts. AB - BACKGROUND: The Viscous Criterion (VC) is an experimental measure developed by the automotive industry to assess injury risks for high-speed impacts. The Blunt Criterion (BC) is a prospective measure developed by the Department of Defense to predict injury from blunt projectiles. METHODS: The range of applicability of BC was extended and compared with VC for its ability to assess injury risk using published cadaver and animal data. Department of Defense projectiles were 0.05 to 0.43 kg mass at velocities up to 86 m/s. VC data were generated from impacts with 1.75 to 23.4 kg at 3.6 to 10.2 m/s. Chest and abdominal injuries ranged from Abbreviated Injury Scale scores of 1 to 6. RESULTS: Both criteria correlated very well with the experimental data, demonstrating correlation coefficients of R = 0.84 to 0.96. The correlation between VC and BC was R = 0.99. Logistic probability curves were derived to predict blunt impact injuries of Abbreviated Injury Scale scores of 1 to 6 for the chest and abdomen. CONCLUSION: BC and VC are virtually identical in their ability to assess blunt and ballistic impact injury risks. They are different measures of impact energy absorbed by the body. One is predictive using input parameters, and the other measures the impact response of the body. PMID- 15128141 TI - Urgent and emergent thoracotomy for penetrating chest trauma. AB - BACKGROUND: Resuscitative thoracotomy (TCY) after trauma has an overall dismal survival rate, yet patients with isolated penetrating chest wounds have the best chance of meaningful recovery. Although the major factor in outcome is presenting physiology, the site of the TCY may influence survival, with the operating room offering a superior environment to the emergency room. PURPOSE: The purpose of this study was to evaluate the impact of location of TCY on outcome after penetrating chest injury. METHODS: This was a multicenter study of patients admitted with either stab (SW) or gunshot wound (GSW) to the chest, with systolic blood pressure < or = 90 mm Hg, and who underwent TCY within 60 minutes of arrival. Time to TCY, Injury Severity Score, location of TCY (emergency room, operating room, or resuscitation room), and detectable systolic pressure at admission were among the factors studied. RESULTS: Over a 4-year period, 78 SW and 140 GSW victims underwent TCY. GSW victims had greater Injury Severity Scores (39.4 +/- 23.1 for GSW vs. 27.2 +/- 15.7 for SW, p < 0.001) and mortality (69% for GSW vs. 37% for SW, p < 0.001). No parameter studied was found to be significantly associated with survival after SW. After GSW, survival was 13.5 times more likely if TCY was performed in the resuscitation room (confidence interval, 3.3-54.6) and 22 times more likely if it was performed in the operating room (confidence interval, 6.7-73.7). CONCLUSION: Although patient selection is the primary factor determining outcome, there may be an independent benefit for performing TCY after GSW in a specialized resuscitation room or the operating room. PMID- 15128142 TI - Knowledge, beliefs, and practices concerning seat belt use during pregnancy. AB - OBJECTIVE: This study seeks to evaluate the comprehension of and attitudes toward proper restraint use among women attending prenatal care clinics. METHODS: Four hundred and fifty women were asked to complete a survey during prenatal care visits at county health department clinics; the response rate was 92.0%. Women were asked to provide demographic information and report their frequency and knowledge of proper automobile restraint use. RESULTS: Nearly all subjects (95.4%) either maintained or increased their pre-pregnancy frequency of restraint use. Three-hundred (72.5%) subjects demonstrated that they wore their restraints in the correct location, with women who wore restraints more frequently being more likely to report correct placement. Two-hundred and forty-nine (60.1%) of women reported that restraints would protect their baby if they were involved in a collision, while 48 (11.6%) thought the restraints would cause injury to their baby, and 153 (37.0%) were unsure. Women who reported that restraints would protect them and their baby if involved in a collision were significantly more likely to report always wearing restraints compared with those who were unsure or had negative perceptions of restraints (84.4% vs. 64.6%; p < 0.0001). The most commonly reported reasons for lack of restraint use were lack of comfort (52.8%) and forgetfulness (42.5%). Only 36.9% percent of women reported receiving information regarding restraint use during their current pregnancy. CONCLUSIONS: Many gravid women lack information regarding proper seat belt use and their role in injury prevention. Consequently, the frequency of seat belt use and its correct placement are negatively impacted. Health care workers should take an active role in educating pregnant gravid women about proper restraint use. PMID- 15128143 TI - The effect of nondiscretionary concealed weapon carrying laws on homicide. AB - BACKGROUND: Historically, the carrying of concealed firearms has been either substantially restricted or prohibited outright. Over the past two decades, laws making it easier for civilians to obtain permits allowing them to carry concealed weapons legally have proliferated throughout the United States. This study investigates the effect of such changes in state laws on state homicide rates. METHODS: Pooled cross-sectional time-series data (1979-1998) for 50 states and Poisson regression methods were used to estimate the effect of changes in state laws on homicide rates. RESULTS: No statistically significant association exists between changes in concealed weapon laws and state homicide rates. This finding is consistent across all models. CONCLUSIONS: The current findings are consistent with those of other published studies indicating that nondiscretionary concealed weapon laws are not associated with significant increases or decreases in homicide. PMID- 15128144 TI - The relation between trauma center outcome and volume in the National Trauma Databank. AB - BACKGROUND: Regionalization of trauma care services aims to improve outcomes by limiting trauma care delivery to a select group of dedicated trauma centers. However, the evidence linking trauma center volume and outcome is not conclusive. The objective of this study was to examine the volume-mortality relation for patients with severe trauma in the National Trauma Databank. METHODS: This study was based on data for adult patients 18 years of age or older in the National Trauma Databank with an Injury Severity Score (ISS) of 15 or more who sustained either blunt or penetrating trauma. The main outcome measure was in-hospital survival as a function of trauma center volume. Logistic regression modeling was used to analyze the relation between survival and hospital volume for patients sustaining either severe blunt or severe penetrating trauma. RESULTS: For the blunt trauma cohort, model diagnostics showed that the single highest-volume center was an outlier. After exclusion of the patients from this center, no association could be demonstrated between trauma volume and outcome (p = 0.465) for blunt trauma. A separate multivariate analysis of patients with penetrating trauma also could not demonstrate a significant volume-mortality association (p = 0.919). Both regression models exhibited excellent discrimination and acceptable calibration. CONCLUSION: The findings of this study do not support the position that higher trauma center volumes are associated with improved survival. The implication of this study is that the hospital volume criteria established by the American College of Surgeons may need to be reexamined. PMID- 15128145 TI - Complicated management of a traumatic aortoesophageal fistula in a patient with a right-sided aortic arch. PMID- 15128146 TI - Traumatic tricuspid insufficiency after horse kick. PMID- 15128147 TI - Ventricular septal rupture in an amateur rodeo rider. PMID- 15128148 TI - Delayed recognition of lumbosacral fracture dislocations in the multitrauma patient: the triad of transverse process fractures, unilateral renal contusion and lumbosacral fracture dislocation. PMID- 15128149 TI - Closed reduction of a symphysis fracture in a 2-month-old infant: treatment considerations. PMID- 15128150 TI - A comminuted talar body fracture osteosynthesized with bioabsorbable screws: a case report. PMID- 15128151 TI - Trapezoid and scaphotrapezial dislocation. PMID- 15128152 TI - Craniofacial metal bolt injury: an unusual mechanism. PMID- 15128153 TI - Nocturnal dyspnea in a young adult male patient: a typical case of an unrecognized traumatic rupture of the diaphragm. PMID- 15128154 TI - Positive predictive value of rib fractures as an indicator of nonaccidental trauma in children. PMID- 15128155 TI - Out-of-hospital rapid-sequence intubation for trauma patients. PMID- 15128156 TI - Surgical abdominal decompression on relieving intractable ICH is of considerable general interest. PMID- 15128157 TI - Recent advances in occupational health research in Korea. AB - Korea has a short history in research on occupational health like as short history of industrialization. During last four decades, however, Korea has experienced what developed countries have experienced for more than a hundred year. Research on occupational health in Korea has also drastically developed. Since industrialization in 1970s, many workers were exposed to hazardous working environment and suffered from occupational accidents and diseases. The main research topics were pneumoconiosis, noise-induced hearing loss and some chemical poisoning. However, improving working condition was not the top priority until the late 1980s. Carbon disulfide poisoning gave a big impact to the society. It made the government take many actions to improve working condition through regulation, enforcement, supporting academia, raising research fund, and establishing a research institute. Recently, classical occupational diseases have decreased and the interest from researchers has also reduced. Many claims for stress-related cardio-cerebrovascular diseases brought much concern and research on job stress. Work-related musculoskeletal disease became a major issue. Many workers are interested in quality of life, such as health promotion. Therefore, research on health promotion, job stress, and psychological problem from work organization would be the main research topics in the future, although research on occupational diseases, such as asthma, cancer and various diseases caused by chemicals are still attractive to researchers. PMID- 15128158 TI - Perspectives on new chemical regulatory framework and research direction in Korea. AB - In response to new information about the health risk of chemical and concern expressed by academia, labor, and industry, chemical regulatory framework under Industrial Safety and Health Act was completely restructured in Year 2003 to be more systematic and to meet the particular needs in Korea. The history and the current status of chemical regulatory framework in the Republic of Korea are reviewed. PMID- 15128159 TI - High signal intensities on T1-weighted MRI as a biomarker of exposure to manganese. AB - Increased signal in T1-weighted images was observed in the experimental manganese (Mn) poisoning of the non-human primate and a patient with Mn neurointoxication. However, our study showed that the increased signals in magnetic resonance images (MRI) were highly prevalent (41.6%) in Mn-exposed workers. Especially 73.5% of the welders showed increased signal intensities. Blood Mn concentration correlated with pallidal index. These changes in MRI tend to disappear following the withdrawal from the source of Mn accumulation, despite permanent neurological damage. Thus increased signal intensities on a T1-weighted image reflect exposure to Mn, but not necessarily manganism. Our study also showed that the concentration of Mn required to produce increased signal intensities on MRI is much lower than the threshold necessary to result in overt clinical signs of manganism. Increased signal intensities in the globus pallidus were determined by Mn accumulation in the animal experiment. All these results strongly suggest that signal intensities in T1-weighted MRI reflect a target site dose in a biologically-based dose-response model. At which increase of signal intensity, the progression of manganism from Mn exposure occurs, however, remains to be solved. PMID- 15128160 TI - Current status of the occupational health and safety countermeasures in Beijing, China. AB - This paper describes the current status of occupational health and safety countermeasures in Beijing, China. It includes the network and organization of occupational health, occupational health personnel, the refitting of industrial structures, hazardous agents at workplaces, the classification and incident rates of occupational diseases. The management of occupational health and safety, new challenges and opportunities for occupational health and safety are also discussed. Countermeasures for occupational health and safety have been formulated by the government of Beijing. At present, they are being implemented and enforced by: (1) strengthening the management of occupational hygiene; (2) encouraging and supporting employment units establishing the occupational health quality management system; (3) promoting research on the prevention and treatment of occupational diseases; (4) improving the transparency in occupational health and safety; (5) motivating the government to improve regulation and standards and (6) developing external occupational health and safety aid programs. PMID- 15128161 TI - Occupational health research in Taiwan. AB - This article gave a brief introduction of population, labor force, general status of occupational safety and health in Taiwan. Statistics of occupational injuries and health disorders, laws and regulations relevant to occupational health were also covered. Research activities driven by universities, research institutes, society/association were provided. Two multi-lateral collaborative research examples were presented: an intoxication outbreak-initiated CS2 study and an information-demand-motivated 2-methoxy-ethanol study. Industrial hygienists, engineers, epidemiologists, and occupational physicians from both universities and research institutes, governmental agencies, and from non-profit R&D organizations and academic associations were involved in these researches, presenting a promise that integrated collaboration of inter-disciplinary specialty cooperated with governmental participation could benefit not only academic achievement, governmental policy-makeup, but also to the employees themselves. PMID- 15128162 TI - Occupational health and safety situation and research priority in Thailand. AB - The aims of this article are to review the situation of occupational health and safety and to describe research priority in this field in Thailand. Thailand is one of newly industrialized countries in Southeast Asia. Approximately half of the working population is still in agriculture. The data from Thai Workmen's Compensation Fund showed that incidence rate of occupational injuries and diseases was 3-4% each year. Almost were occupational injuries from various accidents in workplaces. At least 3 relevant governmental agencies, including Ministry of Labour, Ministry of Public Health, and Ministry of Industry, are responsible in occupational health and safety in the country. Nowadays, those agencies collaborate and develop projects and activities to prevent and control of the problems. Because of lack of staff and other resources, research priority is needed and has been developed recently. The framework of research needed focuses on research and development such as how to improve occupational health and safety management at all levels, setting up and development of standard guidelines for health and environmental assessment, and implementation of suitable control measures in workplaces. Finally, improvement of research system in the country is essential to cope with new occupational health problems in the near future. PMID- 15128163 TI - Occupational health research in India. AB - India being a developing nation is faced with traditional public health problems like communicable diseases, malnutrition, poor environmental sanitation and inadequate medical care. However, globalization and rapid industrial growth in the last few years has resulted in emergence of occupational health related issues. Agriculture (cultivators i.e. land owners + agriculture labourers) is the main occupation in India giving employment to about 58% of the people. The major occupational diseases/morbidity of concern in India are silicosis, musculo skeletal injuries, coal workers' pneumoconiosis, chronic obstructive lung diseases, asbestosis, byssinosis, pesticide poisoning and noise induced hearing loss. There are many agencies like National Institute of Occupational Health, Industrial Toxicology Research Centre, Central Labour Institute, etc. are working on researchable issues like Asbestos and asbestos related diseases, Pesticide poisoning, Silica related diseases other than silicosis and Musculoskeletal disorders. Still much more is to be done for improving the occupational health research. The measures such as creation of advanced research facilities, human resources development, creation of environmental and occupational health cells and development of database and information system should be taken. PMID- 15128164 TI - Drudgery, accidents and injuries in Indian agriculture. AB - The Indian farming employs 225 million workforce to cover 140 million hectares of total cultivated land. In spite of rapid farm mechanization (e.g., 149 million farm machinery), the vast resource-poor family farming has primary dependence on traditional methods (e.g., 520 million hand tools and 37 million animal-drawn implements are in operation). The work drudgery, the traumatic accidents and injuries are the major concerns to examine options for ergonomics intervention and betterment of work in crop production activities. This review summarizes human energy expenditure in crop production activities, to assess the job severity, tools and machinery, and formulate the basis to reorganize work and work methods. While the farm mechanization is more in the northern India, the accidents were more in the villages in southern India. On average of the four regions, the tractor incidents (overturning, falling from the tractor, etc.) were highest (27.7%), followed by thresher (14.6%), sprayer/duster (12.2%), sugarcane crusher (8.1%) and chaff cutter (7.8%) accidents. Most of the fatal accidents resulted from the powered machinery, with the annual fatality rate estimated as 22 per 100,000 farmers. The hand tools related injuries (8% of the total accidents) were non-fatal in nature. In spite of the enactment of legislation, the shortcomings in production and monitoring of the machinery in field use may be responsible for the high rate of accidents (e.g., 42 thresher accidents/1,000 mechanical threshers/year in southern India). Due to the lack of technical capability of the local artisans, adhering to safety and design standards is impractical to the implements fabricated in the rural areas. The analysis emphasizes that the effective safety and health management may be possible through legislative enabling of the local infra-structure, such as block development authority and primary health services, to permeate occupational health and safe work practices in the farming sector. PMID- 15128165 TI - Pulmonary asbestos and non-asbestos fiber concentrations in autopsied inhabitants in Pohang, Korea. AB - To establish reference values for pulmonary asbestos and non-asbestos fiber concentrations in rural Korean residents, and their comparison with those of urban Korean residents and the Japanese, autopsied lung samples from 22 subjects (20 males and 2 females), in Pohang, without known occupational asbestos exposure histories, were analyzed for asbestos and non-asbestos fibers, using transmission electron microscope equipped with an energy dispersive X-ray analyzer. Chrysotile was the major fiber type found in the lungs of the subjects. The residents in Pohang had significantly lower asbestos and non-asbestos fiber concentrations than the Korean urban residents. The Koreans had significantly lower asbestos and non-asbestos fiber concentrations than the Japanese. PMID- 15128166 TI - Airborne asbestos and non-asbestos fiber concentrations in non-occupational environments in Korea. AB - Both airborne asbestos and non-asbestos fiber concentrations were evaluated in Korean non-occupational environments. The airborne fiber concentrations were analyzed in 96 air samples, from 48 different points, by transmission electron microscopy, with energy-dispersive X-ray analysis. The geometric means of the airborne asbestos and non-asbestos fiber concentrations were 0.62 and 67.86, and 0.30 and 17.47 fibers/liter in urban and rural areas, respectively. There were significant differences in both the airborne asbestos and non-asbestos fiber concentrations between the urban and rural areas (p<0.05). The geometric means of airborne asbestos and non-asbestos fiber concentrations were 0.67 and 37.93, and 0.27 and 30.67 fibers/liter at the points less than 10 m and more than 30 m away from highways, respectively. The airborne asbestos concentrations were significantly higher at the points less than 10 m away than at the points more than 30 m away from highways (p<0.01). PMID- 15128167 TI - Quantitative assessment of bone marrow cellularity by magnetic resonance imaging in workers with long-term exposure to solvents. AB - This study was conducted to develop a noninvasive method of bone marrow cellularity evaluation in solvent-exposed painters. Six painters with hypocellular marrow and 132 referents were examined using magnetic resonance imaging (MRI). Full examination of the peripheral blood and bone marrow biopsy was done on each patient. Signal indices were calculated from the signal intensities measured at the vertebral bodies from T12 to S1 and on the paraspinal muscles on both the T1- and the T2-weighted image (T1WI and T2WI). Bone marrow cellularities of the painters were between 20.3% and 33.6%. Signal indices at T1WI were greater in the hypocellular marrow cases compared to those of the referents (p<0.05, p<0.01) and were significantly higher in older women compared with men (p<0.05). After adjusting for age and gender, the signal index of cases at S1 of T1WI was higher than that of the referents by 0.364. Five of the six cases had signal index at S1 of T1WI higher than the mean +1 standard deviation for the same age group and gender. MRI signal indices can be used as a useful indicator of bone marrow cellularity in a high-risk population after adequate adjustment. PMID- 15128168 TI - A pilot study of reference vibrotactile perception thresholds on the fingertip obtained with Malaysian healthy people using ISO 13091-1 equipment. AB - The purpose of this paper is to clarify the reference vibrotactile perception thresholds (VPT) for healthy people in Malaysia. The measurement equipment standard, ISO 13091-1, of the vibrotactile perception thresholds for the assessment of nerve dysfunction and the analysis and interpretation of measurements at the fingertips standard, ISO 13091-2, were published in ISO/TC108/SC4/WG8 on 2001 and 2003 individually. In the ISO 13091-2 standard, the reference VPT data were obtained from few research papers. Malaysian people's VPT data don't include to this standard. In Malaysia, when the VPT is using to diagnose of the hand-arm vibration syndrome, the reference VPT data need to compare with the worker's ones. But, Malaysia does not have the reference VPT data yet. So, in this paper, the VPT was measured by using ISO 13091-1 standard equipment to obtain the reference data for Malaysian people. And these data were compared with the ISO reference data on the ISO 13091-2 standard. From the comparison of these data, it was clear that the Malaysian healthy people's VPT data were consistent with the reference data of the ISO 13091-2 standard. PMID- 15128169 TI - Participatory approaches to improving safety and health under trade union initiative--experiences of POSITIVE training program in Asia. AB - The participatory, action-oriented training program in occupational safety and health named POSITIVE (Participation-Oriented Safety Improvements by Trade Union InitiatiVE) was established in Pakistan and extended to other countries in Asia. The steps taken in the development of the POSITIVE program included collecting local good examples in safety and health, developing an action-checklist, testing a participatory training program, and conducting follow-up activities to examine local achievements. Training manuals were compiled to provide workers with the practical, easy-to-understand information on safety and health improvements and on the positive roles of trade unions. Trade union trainers trained in the methodology conducted serial POSITIVE training workshops in Pakistan and then in Bangladesh, Mongolia, Nepal, the Philippines and Thailand and recently in China. These workshops resulted in many low-cost improvements at the workplace level. These improvements were carried out in the technical areas of materials handling, workstations, machine safety, physical environment, and welfare facilities. The trade union networks have been vital in reaching an increasing number of grass root workplaces and in expanding the program to other countries. This included the visits to Mongolia and Thailand of Pakistani trade union trainers to demonstrate the POSITIVE training. The participatory training tools used in the POSITIVE program such as the action checklist and group discussion methods were commonly applied in different local situations. Participatory approaches adopted in the POSITIVE program have proven useful for providing practical problem solving measures based on the local trade union initiative. PMID- 15128170 TI - Toxicity of cyanoacrylate adhesives and their occupational impacts for dental staff. AB - Cyanoacrylate (CA) and its homologues have a variety of medical, dental and commercial applications as adhesives. The increasing use of CA in dentistry, particularly as an adhesive and sealing glue, has raised concerns regarding its potential toxicity in humans. Reported toxicity of CA is uncommon in the dental workplace, but may manifest as conditions such as urticaria, contact dermatitis and other dermatoses. Dental staff using CA adhesives should avoid direct contact with CA and use appropriate personal protective measures. Maintaining higher levels of humidity, optimizing room ventilation and using special air conditioning filters in the working environment may be useful in minimising the toxicity of volatile CA adhesives. PMID- 15128171 TI - Standardized indices of mortality among persons with spinal cord injury: accelerated aging process. AB - We conducted this study to compare survival rates and morbidity of persons with spinal cord injury (SCI) versus general population, and to clarify the risk of SCI persons. The subjects of this study were 960 men with SCI who had been accommodated in the eight Labor Accident Rehabilitation Centers in Japan during the period of 1965-1995. The surveyed items were the year of birth, the year of injury, level of spinal cord injury and survival status. The classification of cause of death was taken from ICD-10. The cumulative survival rate by life table method was calculated. In order to compare the risks of each cause of death in SCI persons with general population, cause-specific standardized mortality ratio (SMR) was examined. The leading cause of death was malignant neoplasms at 28 persons, and SMR (general population=100) was 184, followed by the circulatory system disease, external cause (including suicide) and the genitourinary system disease. In the subgroups of malignant neoplasms, the SMR was 6,619 for cutaneous carcinomas and 1,482 for bladder carcinomas (p<0.01). Thus aging-related diseases which had close correlation with lifestyle and environment were the major cause of deaths in SCI persons. PMID- 15128172 TI - Comparative investigation of several sperm analysis methods for evaluation of spermatotoxicity of industrial chemical: 2-bromopropane as an example. AB - Reproductive toxicity of 2-bromopropane (2BP), a substitute for ozone layer depleting chloro-fluorocarbon, was found among the workers in an electronics factory in Korea in 1995. Furthermore the importance of testicular toxicity has been realized since the problem of endocrine disruptors arose all over the world, but manual methods must rely on subjective assessment. Recently, computer assisted sperm analysis (CASA) was proposed but this system requires vast investment. We then investigated the applicability of the MTT method with a microplate and sperm quality analyzer (SQA) as simple, rapid, and economic instrumental methods for the examination of sperm quality in rats, comparing it with the manual microscopic method and CASA. Epididymal fluid derived from male F344/N Slc (Fischer) rats intraperitoneally injected with 2BP in the dose range of 125-1,000 mg/kg/d twice a week (total 8 times) were examined by these methods as a model experiment. Sperm count measured by the manual method and CASA in the epididymal fluid, absorbance by the MTT method and sperm motility index value by the SQA method were significantly lower in the 2BP 1,000 mg/kg administered group than in the control group. This result suggests that the MTT method can detect oligospermia. With the microplate and microplate reader, the efficiency of detection becomes much better. Sperm analyses by the MTT method with the microplate reader and the SQA method are available for reproductive toxicity study in rats. PMID- 15128173 TI - A field method for sampling toluene in end-exhaled air, as a biomarker of occupational exposure: correlation with other exposure indices. AB - A sensitive and rapid method for the determination of toluene in exhaled air is described. We have developed a device for direct breath sampling consisting of a sampler inserted into an empty 58 mL glass vial closed by a Teflon rubber septum. The sorbent cartridge functions as a diffusive sampler and employs a Tenax resin (300 mg, 35/50 mesh) to trap volatile organic compounds from the exhaled air. End exhaled air is collected "in field" by removing the septum from the vial, by forcibly exhaling into the device through a suitable Teflon tube, and then by sealing the bottle quickly. Environmental toluene levels ranged from 13 to 191 mg/m3, while the concentrations of the solvent in alveolar air, in blood and urine ranged from 159 to 3354 ng/L, from 3.6 to 53.5 microg/L, and from 8.7 to 142.4 microg/L respectively. The correlation coefficients (r) of biological measurements towards environmental toluene levels were 0.822, 0.850 and 0.846 for alveolar air, blood and urine samples, respectively. The breath sampler allowed the rapid and non-invasive collection of data on elimination of toluene. PMID- 15128174 TI - Familial mesothelioma of the pleura--a report of 40 cases. AB - A survey of 610 pleural mesotheliomas disclosed 40 familial cases. The diagnosis was histologically based in 39 cases, and confirmed by necropsy in 30. Occupational data were collected from the patients or from their relatives by personal interviews. Routine lung sections were examined for asbestos bodies in 32 cases. In 15 cases asbestos bodies were isolated after chemical digestion of lung tissue. Familial mesotheliomas included 31 men and 9 women (age range 44-93 yr, mean 70.7, median 71.0). In 15 families there were blood relations between (or among) the members involved. All the patients had been exposed to asbestos, mostly in the shipyards. Asbestos bodies were found on routine lung sections in 27 cases. Asbestos bodies after isolation ranged from 70 bodies to about 900,000/g dried lung tissue. Latency periods (time intervals between first exposure to asbestos and diagnosis) ranged between 25 and 70 yr (mean 52.0, median 54.0). The occurrence of mesothelioma among subjects with blood relations suggests that genetic factors might play a role in determining the susceptibility to asbestos-related cancer. Familial cases among persons without blood relations raise the question if environmental factors that members of a family share, may act as co-factors in asbestos-related mesothelioma. PMID- 15128175 TI - An accident-risk assessment study of temporary piece rated workers. AB - An occupational injury surveillance study (record study of five years duration) was conducted involving the workers of a fertilizer producing industry in eastern India to assess whether the risk of occupational accidents in temporary piece rated workers was higher in comparison to the permanent time rated workers. At the same time, to collect the personal details of the workers who have worked in the industry in the study period, an interview was also conducted. Mean age of temporary piece rated workers and permanent time rated workers were (35.9 +/- 12.5) and (35.3 +/- 11.4) respectively. Distribution of other variables like nature of work, level of education, experience, habits were also very similar between the two worker groups. Accident incidence rate, accident frequency rate and accident severity rate were found to be significantly higher in temporary piece rated workers. This difference was more prominent in case of time-loss accidents than in no time-loss accidents. Relative risk has varied from 2.3 to 18.0 in case of time-loss accidents. In case of no time-loss accidents, it has varied from 1.1 to 2.6. When relative risk is considered after taking both types of accidents together, it has ranged from 1.2 to 3.5. This study concluded that the temporary piece rated workers are more vulnerable to occupational accidents. PMID- 15128176 TI - Exposure to urban pollutants and plasma vasopressin in traffic policemen. AB - OBJECTIVES: The aim of the present study is to investigate whether traffic policemen of a big city exposed to urban pollutants may be at risk of alterations on plasma vasopressin (VP) concentrations compared with a control group. MATERIAL AND METHODS: Out of a population of 395 Municipal Police employees, the subjects with the principal confounding factors were excluded from the study. Traffic policemen and control subjects were matched by sex, age and working life. Plasma VP levels were determined in 82 subjects: 41 traffic policemen (18 men, 23 women) and 41 control subjects (18 men, 23 women). RESULTS: Mean of VP values were found significantly lower in traffic policemen than in the control group both in men (P=0.010) and in women (P=0.015). We found high correlation between VP values and working life in traffic policemen of male and female sex (r=0.7; r=0.6). DISCUSSION: The authors hypothesise an effect on plasma VP levels in traffic policemen exposed to chemical and physical stressors, according to plasma VP levels modifications found by other authors in studies on animals and human subjects. PMID- 15128177 TI - Lower vapor concentrations in solvent workplaces in larger-scale enterprises than in smaller-scale enterprises, and exceptions. AB - The aim of the present study was to investigate the relationship between environmental vapor concentrations in organic solvent workplaces and size of enterprises, and to examine if occupational hygiene conditions were better in larger-scale enterprises. For this purpose, a total of 3,567 solvent workplaces were surveyed for environmental solvent vapor concentrations in 1999 to 2002. The results were classified by the size of enterprises (taking the number of employees as an indicator) and by the type of solvent work. It was observed that the vapor concentration in a typical small-scale enterprise was approximately three times as high as that in the large-scale enterprise, although the administrative control levels were not exceeded in general. The proportion of testing and research work (with less use of organic solvents and thus low vapor concentrations) among all solvent workplaces was higher in large-scale enterprises than in small-scale enterprises. The vapor concentrations in this type of workplace did not differ however irrespective of the enterprise size. Further comparison in various types of solvent workplaces disclosed similar size dependent difference in workplaces for degreasing, cleaning, wiping, printing, and surface coating. In painting and solvent-drying work, in contrast, there was no difference in environmental concentration regardless of the size of enterprises, possibly because environmental improvement of this type of workplaces was technically more difficult than others. Thus, it was concluded that large-scale enterprises generally had better control of work environments than small-scale enterprises, with possible exceptions of painting work and solvent-drying work. PMID- 15128178 TI - Psychometric evaluation of the Chinese (mainland) version of Job Content Questionnaire: a study in university hospitals. AB - To evaluate the psychometric properties of the Chinese (mainland) version of Job Content Questionnaire (JCQ), this validation study was conducted in university hospitals among 193 men and 581 women health care workers. The Cronbach's alpha coefficients for decision latitude, psychological job demands and social support ranged from 0.56 to 0.82, which indicated that psychological job demands had lower internal consistency. The decision latitude displayed a negative correlation with psychological job demands and a positive correlation with social support. Exploratory factor analysis showed 3 meaningful factors that could explain the 3 theoretical dimensions of job strain model. Apart from some uncertainty over psychological job demands particularly, it is concluded that the Chinese (mainland) version of JCQ is a reliable and valid instrument for measuring job stressors, and is applicable to Chinese working population. PMID- 15128179 TI - Effects of night work on urinary excretion rates of 6-sulfatoxymelatonin, norepinephrine and estriol in pregnant women. AB - It has been suggested that shift work, night work in particular, affects worker's psychophysical health. However, the effects of night work on the health of pregnant women are not physiologically well elucidated. The effects of night work on the biological function of pregnant women were studied in the present study. Three pregnant and six non-pregnant nurses that engaged in fast-rotating shift system cooperated for the study. The 24-h urine samples were collected in two time frames, daytime (07:00 to 23:00) and nighttime (23:00 to 07:00) on the day shift, the night shift and the days off. Urinary concentrations of 6 sulfatoxymelatonin and norepinephrine were measured by enzyme-linked immunosorbent assay and estriol by radioimmunoassay. The excretion profiles of urinary 6-sulfatoxymelatonin in the pregnant and the non-pregnant women were altered on the night shift, which might relate the derangement of circadian rhythm of melatonin secretion. The urinary norepinephrine level during the night work was considerably higher in the pregnant women, which indicated the presence of higher physical burden in them due to the night work. The urinary estriol level was not significantly affected by shift, day/night and individual factors, and the effect of night work on urinary estriol level of the pregnant women remained uncertain. The results of the present study suggested that the night work of the pregnant shift workers should be much more relieved. PMID- 15128180 TI - Quantitative analysis of tridymite and cristobalite crystallized in rice husk ash by heating. AB - The quantities of two forms of crystalline silica, tridymite and cristobalite, in heated rice husk ash (RHA) samples were determined by X-ray diffraction (XRD) and chemical methods. Two RHA samples, containing 93% SiO2 and 2-3% K2O, were prepared from charcoaled rice husk products and heated to above 900 degrees C. The crystalline silica made up over 60-80% of the total silica in the heated RHA samples based on the XRD analysis. The crystalline phases in the two samples were somewhat different: The sample heated in the temperature range of 900 to 1,200 degrees C contained 52-62% cristobalite and 10-17% tridymite, but the other sample heated at a comparable temperature, above 1,100 degrees C, contained 46 66% tridymite and 37-16% cristobalite. Based on a correlation of lower tridymite crystallization temperature with higher potassium content, it was concluded that higher potassium levels were responsible for this difference. The pyrophosphoric acid analysis did not give exact results in the evaluation of total crystalline silica content in these RHA samples. As the combustion of rice husk was considered to cover the demands for energy and silica resource in Asian countries, cristobalite and tridymite crystallized in RHA by burning of rice husk should be assessed precisely by XRD analysis and the airborne dust in relevant workplace be controlled. PMID- 15128181 TI - Occupational health nurses' participation in health committee surveyed in Yamaguchi prefecture, Japan. AB - This survey was conducted to know the present conditions of occupational health nurses (OHNs) activity related to health committees (HCs) in Japan. Questionnaires that included items related to duties of the OH physician (OHP), those of the OHN, and their mutual duties within the HC, were mailed to 41 companies employing OHNs and questionnaires from 18 companies were analyzed. Comparison of the frequency of OHN attendance at Health Committee Meetings (HCMs) revealed that 33.3% of OHNs attended the HCs when their companies employed full time OHPs and 83.3% attended when their companies employed part-time OHPs. In a question about the OHN's opportunity to deliver a speech, give a report or make a presentation at their HCMs, 16.7% of OHNs in companies with full-time OHPs and 66.7% of OHNs in companies with part-time OHPs reported they had such opportunity. In companies with part-time OHPs, 50.0% of the OHNs reported that they were asked for their opinions at the HCMs, but OHNs at companies employing full-time OHPs were not asked for their opinions. It was considered that in the future, OHNs, particularly those working with a part-time OHP, will have an important role in the HC and developing OH services for their companies. PMID- 15128182 TI - Development of action checkpoints for comfortable computer work. AB - We developed a manual including a checklist format for undertaking measures to prevent fatigue in visual display terminals (VDT) workers. With this manual, problems related to VDT work can be recognized by using checklists which allow for self-evaluation by the workers. The manual helps the workers to consider measures for improvement by themselves and will contribute to better occupational health education. PMID- 15128183 TI - Three thiamine analogues differently alter thiamine transport and metabolism in nervous tissue: an in vivo kinetic study using rats. AB - Thiamine (T) analogues pyrithiamine, oxythiamine or amprolium in amounts 10-1000 times higher than labelled T, were i.p. injected into rats together with 14C-T (30 microg; 46.25 KBq). The radioactivity associated with T and its phosphoesters in the plasma and cerebral cortex, brainstem, cerebellum, and sciatic nerve were determined at time intervals from 0.25 to 240 h from injection. The experimental data obtained were processed with a mathematical compartmental model that calculated the fractional rate constants. These are the amount of content in a given compartment that is replaced in 1 h and expressed in per hour. The results showed that all three analogues inhibited thiamine entry from plasma. Instead, oxythiamine enhanced T phosphorylation to T pyrophosphate (TPP); amprolium and oxythiamine enhanced TPP dephosphorylation to monophosphate (TMP); pyrithiamine reduced TPP dephosphorylation and TMP formation, while none of the analogues modified TMP dephosphorylation to T. In conclusion, in living rats, the action of T analogues was much more complex than could be expected from their structure and action in vitro. PMID- 15128184 TI - Lactate stress testing by bedside lactate determination. AB - Lactate determination for the lactate-stress-test (LST) by means of a bedside method has not been performed. Serum lactate was determined by means of the Ektachrome Clinical Chemistry Slide (LST1) and bedside by means of the Acutrend Lactate (LST2) once before, 3 times during, and once after a 15-min, constant 30 W workload on a bicycle in 20 controls, 21 disease controls, and 22 patients with mitochondriopathy (MCP). Lactate's upper reference limits at rest, 5, 10, 15 min after starting, and 15 min after finishing the exercise were 2.0, 1.9, 1.9, 1.8, and 1.5 mmol/L for the LST1 and 2.5, 2.9, 2.5, 2.7, and 2.0 mmol/L for the LST2. The LSTI (LST2) was abnormal in 17 (18) MCP patients, 7 (3) disease controls, and none (none) of the healthy subjects. The sensitivity of the LST1 (LST2) for MCP patients was 77% (82%). The specificity of the LST1 (LST2) was 67% (86%). Sensitivity and specificity of the LST2 are higher than that of the LST1. The LST2 can thus replace the LST1, since it is also easier to handle, quick, reliable, and cheaper. PMID- 15128186 TI - Joint manipulation curricula in physical therapist professional degree programs. AB - STUDY DESIGN: Descriptive observational survey. OBJECTIVE: To describe the status of joint manipulation curricula within physical therapist professional degree programs in the United States. BACKGROUND: Studies have described the evolution of manual therapy curricula, including spinal and extremity joint mobilization, in physical therapist professional programs, but minimal information exists related to joint manipulation curricula. METHODS AND MEASURES: Primary faculty members responsible for teaching manual therapy curricular content at the 199 physical therapist professional degree programs located in the United States recognized by the Commission on Accreditation in Physical Therapy Education were asked to participate in this project. The survey documented joint manipulation curricula, faculty qualifications, attitudes and experience, and programs' future plans for teaching manipulation. RESULTS: Of the 116 programs responding to our survey, 87 (75%) currently include joint manipulation in their curriculum or plan to soon include such content in their curriculum. Of the programs currently teaching joint manipulation, 75% taught it as part of a required integrated clinical science course. Faculty teaching manipulation content appear to be well qualified and are in clinical practice an average of 12 hours per week. The programs currently not teaching joint manipulation reported reasons, including belief that it was not an entry-level skill (45%), lack of time (26%), lack of qualified faculty (71%), and perceived lack of scientific evidence regarding efficacy (7%). CONCLUSIONS: Of the responding professional degree programs, 75% are either currently teaching joint manipulation or soon plan to do so. Our research may serve as a benchmark for faculty to assess existing manual therapy curricula and as a guide for developing curricula in new or existing physical therapy programs. PMID- 15128185 TI - In vitro homocysteine inhibits platelet Na+,K+-ATPase and serum butyrylcholinesterase activities of young rats. AB - Homocystinuria is an inborn error of sulphur amino acid metabolism, resulting in accumulation of tissue homocysteine. This disease is characterized predominantly by vascular and nervous system dysfunction. In the present study we investigated the in vitro effects of homocysteine, the main metabolite accumulated in homocystinuria, on platelet Na+,K+-ATPase and serum butyrylcholinesterase (BuChE) activities of young rats. Platelet and serum of 29-day-old Wistar rats were incubated in the absence (control) or presence of homocysteine (0.01-0.5 mM). Results showed that Na+,K+-ATPase and BuChE activities were significantly inhibited by homocysteine. It is proposed that inhibition of Na+,K+-ATPase and BuChE activities might be one useful peripheral marker for the neurotoxic effects of homocysteine. PMID- 15128187 TI - Identification of a fibular fracture in an intercollegiate football player in a physical therapy setting. PMID- 15128188 TI - The evaluation of change in pain intensity: a comparison of the P4 and single item numeric pain rating scales. AB - STUDY DESIGN: Prospective observation study. OBJECTIVES: To compare the test retest reliability and longitudinal validity (sensitivity to change) of 2 single item numeric pain rating scales (NPRSs) with a 4-item pain intensity measure (P4). BACKGROUND: Pain is a frequent outcome measure for patients seen in physical therapy; however, the error associated with efficient pain measures, such as the single-item NPRS, is greater than for self-report measures of functional status. Initial evaluation of the P4 suggests that it is more reliable and sensitive to change than the NPRS. METHODS AND MEASURES: Two single-item NPRSs and the P4 were administered on 3 occasions--initial visit (n = 220), within 72 hours of baseline (n = 213), and 12 days following baseline assessment (n = 183)--to patients with musculoskeletal problems receiving physical therapy. Reliability was assessed using a type 2,1 intraclass correlation coefficient. Longitudinal validity was assessed by correlating the measures' change scores with a retrospective rating of change that included patients' and clinicians' perspectives. RESULTS: The test-retest reliability and longitudinal validity of the P4 were significantly greater (P1<.05) than both single-item NPRSs. Minimal detectable change of the P4 at the 90% confidence level was estimated to be a change of 22% of the scale range (9 points) compared to 27.3% (3 points) and 31.8% (3.5 points) for the 2-day NPRS and 24-hour NPRS, respectively. CONCLUSIONS: The findings of this study suggest the P4 is more adept at assessing change in pain intensity than popular versions of single-item NPRSs. PMID- 15128189 TI - Validity of the supraspinatus test as a single clinical test in diagnosing patients with rotator cuff pathology. AB - STUDY DESIGN: Prospective blinded comparison of clinical examination and surgical findings of consecutive patients seen at a tertiary shoulder center. OBJECTIVE: To investigate the validity of the supraspinatus test in diagnosing rotator cuff pathology using arthroscopy or open surgery as reference standards. A positive supraspinatus test was defined as pain for all types of rotator cuff pathology and weakness for full-thickness tears. BACKGROUND: Rotator cuff tenopathy is a very common condition. However, there have been relatively few studies documenting the validity of physical examination for this condition and further investigation of the measurement properties of these tests is warranted. METHODS AND MEASURES: One hundred two consecutive subjects were examined. Fifty subjects, ranging in age between 24 and 79 years (mean age, 50 years; SD, 14.4 years) and composed of 16 females and 34 males, underwent surgery. RESULTS: The sensitivity of the supraspinatus test was 62%, 41%, and 88% for "supraspinatus tendonitis or partial thickness tear," "full-thickness tear," and "large to massive tears," respectively. The specificity values were 54%, 70%, and 70% for the above conditions, respectively. The negative likelihood ratios varied from 0.17 to 0.84, and the positive likelihood ratios varied from 1.35 to 2.93, depending on the presence of pain or weakness. CONCLUSION: Application of the supraspinatus test in isolation is helpful in diagnosing large or massive rotator cuff tears. The change that this test makes in pretest probability of less extensive rotator cuff pathology is insignificant. PMID- 15128190 TI - The effect of low-Dye taping on plantar pressures, during gait, in subjects with navicular drop exceeding 10 mm. AB - STUDY DESIGN: A preintervention and postintervention, repeated-measures experimental design. OBJECTIVES: To investigate the immediate effect of low-Dye taping on peak and mean plantar pressures during gait in subjects with navicular drop exceeding 10 mm. BACKGROUND: Low-Dye taping is commonly used to support the longitudinal and transverse arches of the foot in an attempt to reduce the effects of symptoms associated with excessive pronation. Plantar pressure measurement has been used as an indirect indicator of pronation during gait. METHOD AND MEASURES: The right foot of 60 subjects was tested using the Emed-AT system to obtain plantar pressure values. Subjects performed 6 barefoot walks over the Emed pressure platform while taped and a further 6 walks while untaped. Plantar pressures were recorded. Each footprint obtained was divided into 10 sections or 'masks.' Average peak and mean plantar pressure values (N/cm2) were calculated for both taped and untaped walks for each mask. RESULTS: Paired t tests demonstrated significant changes in peak plantar pressure in 8 of the 10 areas of the foot and significant changes in mean plantar pressure in 9 of the 10 areas of the foot. Low-Dye taping significantly decreased pressure under the heel and the medial and middle forefoot, while increasing pressure under the lateral midfoot and under the toes. A significant decrease in mean plantar pressure was observed under the lateral forefoot, while no significant difference was demonstrated in peak plantar pressure under this area. The area under the medial midfoot demonstrated no significant change in either peak or mean pressure. CONCLUSIONS: Low-Dye taping significantly altered peak and mean plantar pressure values in subjects with navicular drop exceeding 10 mm. In particular, peak and mean plantar pressure increased under the lateral midfoot and under the toes, and decreased under the heel and forefoot, suggesting that a decrease in the amount of pronation occurred. PMID- 15128191 TI - The scanning laser ophthalmoscope--a review of its role in bioscience and medicine. AB - The scanning laser ophthalmoscope (SLO) offers the potential for retinal imaging that is complementary both to that of the fundus camera and also the newly developing technique of optical coherence tomography (OCT). It has the ability to produce rapid images at low light levels using light of specific wavelengths. This permits temporal studies of fluorescent-labelled cells which offer a unique insight into inflammatory processes in the eye. The facility to image with several different wavelengths simultaneously offers the potential for spectral imaging of retinal tissue with the aim of revealing those early changes in tissue perfusion that indicate the onset of retinal disease, so increasing the probability of successful therapy. PMID- 15128192 TI - The potential of optical coherence tomography in the engineering of living tissue. AB - The better repair of human tissue is an urgent medical goal and in order to achieve a safe outcome there is a parallel need for sensitive, non-invasive methods of assessing the quality of the engineered tissues and organs prior to surgical implantation. Optical coherence tomography (OCT) can potentially fulfil this role. The current status of OCT as an advanced imaging tool in clinical medicine, developmental biology and material science is reviewed and the parallels to the engineering of living tissue and organs are discussed. Preliminary data are also presented for a tissue engineering bioreactor with in situ OCT imaging. The data suggest that OCT can be utilized as a real time, non destructive, non-invasive tool to critically monitor the morphology of tissue engineered constructs during their fabrication and growth. PMID- 15128193 TI - Imaging changes in blood volume and oxygenation in the newborn infant brain using three-dimensional optical tomography. AB - Induced haemodynamic and blood oxygenation changes occurring within the brain of a ventilated newborn infant have been imaged in three dimensions using optical tomography. Noninvasive measurements of the flight times of transmitted light were acquired during illumination of the brain by laser pulses at wavelengths of 780 nm and 815 nm. The oxygen and carbon dioxide partial pressures were adjusted through alterations to the ventilator settings, resulting in changes to the cerebral blood volume and oxygenation. Three-dimensional images were generated using the physiologically associated differences in the measured data, obviating the need for data calibration using a separate reference measurement. The results exhibit large changes in absorption coefficient at both wavelengths. Images corresponding to differences in concentrations of oxy- and deoxyhaemoglobin are in qualitative agreement with known physiological data. PMID- 15128194 TI - Breast deformation modelling for image reconstruction in near infrared optical tomography. AB - Near infrared tomography (NIR) is a novel imaging technique that can be used to reconstruct tissue optical properties from measurements of light propagation through tissue. More specifically NIR measurements over a range of wavelengths can be used to obtain internal images of physiologic parameters and these images can be used to detect and characterize breast tumour. To obtain good NIR measurements, it is essential to have good contact between the optical fibres and the breast which in-turn results in the deformation of the breast due to the soft plasticity of the tissue. In this work, a tissue deformation model of the female breast is presented that will account for the altered shape of the breast during clinical NIR measurements. Using a deformed model of a breast, simulated NIR data were generated and used to reconstruct images of tissue absorption and reduced scatter using several assumptions about the imaging domain. Using either a circular or irregular 2D geometry for image reconstruction produces good localization of the absorbing anomaly, but it leads to degradation of the image quality. By modifying the assumptions about the imaging domain to a 3D conical model, with the correct diameter at the plane of NIR measurement, significantly improves the quality of reconstructed images and helps reduce image artefacts. Finally, assuming a non-deformed breast shape for image reconstruction is shown to lead to poor quality images since the geometry of the breast is greatly altered, whereas using the correct deformed geometry produces the best images. PMID- 15128195 TI - Sagittal laser optical tomography for imaging of rheumatoid finger joints. AB - We present a novel optical tomographic imaging system that was designed to determine two-dimensional spatial distribution of optical properties in a sagittal plane through finger joints. The system incorporates a single laser diode and a single silicon photodetector into a scanning device that records spatially resolved light intensities as they are transmitted through a finger. These data are input to a model-based iterative image reconstruction (MOBIIR) scheme, which uses the equation of radiative transfer (ERT) as a forward model for light propagation through tissue. We have used this system to obtain tomographic images of six proximal interphalangeal finger joints from two patients with rheumatoid arthritis. The optical images were compared to clinical symptoms and ultrasound images. PMID- 15128196 TI - Concentration and oxygen saturation of haemoglobin of 50 breast tumours determined by time-domain optical mammography. AB - Using a dual-wavelength (670 nm, 785 nm) time-domain scanning instrument we have recorded optical mammograms of 93 patients suspected of having breast cancer which was subsequently assessed histologically. Among 65 histologically confirmed carcinomas, 54 were detectable in at least one of two optical mammograms recorded of each tumour-bearing breast in craniocaudal and mediolateral projection. Optical mammograms were based on photon counts in selected time windows of measured distributions of times of flight of photons. Optical properties of 50 carcinomas investigated at both wavelengths were derived by modelling the breast as partially homogeneous infinite slab with an embedded spherical inhomogeneity representing the tumour and by calculating the diffraction of photon density waves. In selected cases, additional information about the location of the tumour along the compression direction was used that was obtained from scans at selected offsets between source and detector optical fibres. A correlation plot of haemoglobin concentration and blood oxygen saturation of tumours and healthy tissue shows good separation between both kinds of tissue. The majority of carcinomas exhibited increased total haemoglobin concentration compared to healthy tissue. PMID- 15128197 TI - Optical measurements of absorption changes in two-layered diffusive media. AB - We have used Monte Carlo simulations for a two-layered diffusive medium to investigate the effect of a superficial layer on the measurement of absorption variations from optical diffuse reflectance data processed by using: (a) a multidistance, frequency-domain method based on diffusion theory for a semi infinite homogeneous medium; (b) a differential-pathlength-factor method based on a modified Lambert-Beer law for a homogeneous medium and (c) a two-distance, partial-pathlength method based on a modified Lambert-Beer law for a two-layered medium. Methods (a) and (b) lead to a single value for the absorption variation, whereas method (c) yields absorption variations for each layer. In the simulations, the optical coefficients of the medium were representative of those of biological tissue in the near-infrared. The thickness of the first layer was in the range 0.3-1.4 cm, and the source-detector distances were in the range 1-5 cm, which is typical of near-infrared diffuse reflectance measurements in tissue. The simulations have shown that (1) method (a) is mostly sensitive to absorption changes in the underlying layer, provided that the thickness of the superficial layer is approximately 0.6 cm or less; (2) method (b) is significantly affected by absorption changes in the superficial layer and (3) method (c) yields the absorption changes for both layers with a relatively good accuracy of approximately 4% for the superficial layer and approximately 10% for the underlying layer (provided that the absorption changes are less than 20-30% of the baseline value). We have applied all three methods of data analysis to near infrared data collected on the forehead of a human subject during electroconvulsive therapy. Our results suggest that the multidistance method (a) and the two-distance partial-pathlength method (c) may better decouple the contributions to the optical signals that originate in deeper tissue (brain) from those that originate in more superficial tissue layers. PMID- 15128198 TI - Do shorter wavelengths improve contrast in optical mammography? AB - The detection of tumours with time-resolved transmittance imaging relies essentially on blood absorption. Previous theoretical and phantom studies have shown that both contrast and spatial resolution of optical images are affected by the optical properties of the background medium, and high absorption and scattering are generally beneficial. Based on these observations, wavelengths shorter than presently used (680-780 nm) could be profitable for optical mammography. A study was thus performed analysing time-resolved transmittance images at 637, 656, 683 and 785 nm obtained from 26 patients bearing 16 tumours and 15 cysts. The optical contrast proved to increase upon decreasing wavelengths for the detection of cancers in late-gated intensity images, with higher gain in contrast for lesions of smaller size (<1.5 cm diameter). For cysts either a progressive increase or decrease in contrast with wavelength was observed in scattering images. PMID- 15128199 TI - Combined optimal-pathlengths method for near-infrared spectroscopy analysis. AB - Near-infrared (NIR) spectroscopy is a rapid, reagent-less and nondestructive analytical technique, which is being increasingly employed for quantitative application in chemistry, pharmaceutics and food industry, and for the optical analysis of biological tissue. The performance of NIR technology greatly depends on the abilities to control and acquire data from the instrument and to calibrate and analyse data. Optical pathlength is a key parameter of the NIR instrument, which has been thoroughly discussed in univariate quantitative analysis in the presence of photometric errors. Although multiple wavelengths can provide more chemical information, it is difficult to determine a single pathlength that is suitable for each wavelength region. A theoretical investigation of a selection procedure for multiple pathlengths, called the combined optimal-pathlengths (COP) method, is identified in this paper and an extensive comparison with the single pathlength method is also performed on simulated and experimental NIR spectral data sets. The results obtained show that the COP method can greatly improve the prediction accuracy in NIR spectroscopy quantitative analysis. PMID- 15128200 TI - Three-dimensional cellular-level imaging using full-field optical coherence tomography. AB - An ultrahigh-resolution full-field optical coherence tomography (OCT) system has been developed for cellular-level imaging of biological media. The system is based on a Linnik interference microscope illuminated with a tungsten halogen lamp, associated with a high-resolution CCD camera. En face tomographic images are produced in real time, with the best spatial resolution ever achieved in OCT (0.7 microm x 0.9 microm, axial x transverse). A shot-noise limited detection sensitivity of 80 dB can be reached with an acquisition time per image of 1 s. Images of animal ophthalmic biopsies and vegetal tissues are shown. PMID- 15128201 TI - Advances in broad bandwidth light sources for ultrahigh resolution optical coherence tomography. AB - Novel ultra-broad bandwidth light sources enabling unprecedented sub-2 microm axial resolution over the 400 nm-1700 nm wavelength range have been developed and evaluated with respect to their feasibility for clinical ultrahigh resolution optical coherence tomography (UHR OCT) applications. The state-of-the-art light sources described here include a compact Kerr lens mode locked Ti:sapphire laser (lambdaC = 785 nm, delta lambda = 260 nm, P(out) = 50 mW) and different nonlinear fibre-based light sources with spectral bandwidths (at full width at half maximum) up to 350 nm at lambdaC = 1130 nm and 470 nm at lambdaC = 1375 nm. In vitro UHR OCT imaging is demonstrated at multiple wavelengths in human cancer cells, animal ganglion cells as well as in neuropathologic and ophthalmic biopsies in order to compare and optimize UHR OCT image contrast, resolution and penetration depth. PMID- 15128202 TI - Use of optical coherence tomography in delineating airways microstructure: comparison of OCT images to histopathological sections. AB - An ideal diagnostic system for the human airways should be able to detect and define early development of premalignant pathological lesions, to facilitate optimal curative treatment and prevent irreversible and/or invasive lung disease. There is great need for exploration of safe, repeatable imaging techniques which can run at real-time and with high spatial resolution. In this study, optical coherence tomography (OCT) was utilized to acquire cross-sectional images of upper and lower airways using fresh pig lung resections as a model system. Obtained OCT images were compared with parallel tissue characterization by conventional histological analysis. Our objective was to determine whether OCT differentiates the composite structural layers and inherent anatomical variations along different airway locations. The data show that OCT can clearly display the multilayered structure of the airways. The subtle architectural differences in three separate anatomical locations including trachea, main bronchus and tertiary bronchus were clearly delineated. Images of the appropriate anatomical profiles, with depth of up to 2 mm and 10 microm spatial resolution were obtained by our current OCT system, which was sufficient for recognition of the epithelium, subepithelial tissues and cartilage. In addition, the relative thickness of individual structural components was accurately reflected and comparable to histological sections. These data support OCT as a highly feasible, optical biopsy tool, which merits further exploration for early diagnosis of human airway epithelial pathology. PMID- 15128203 TI - Transversal phase resolved polarization sensitive optical coherence tomography. AB - We present a novel optical coherence tomography (OCT) method to measure backscattered intensity and birefringence properties (retardation and fast axis orientation) and apply it to imaging of human ocular tissue. The method is based on a Mach Zehnder interferometer, on transversal scanning, and on a polarization sensitive two-channel detection. A highly stable carrier frequency is generated by acousto-optic modulators (AOMs). This allows a phase sensitive demodulation by the lock-in technique. Since the recording of individual interference fringes is avoided by this method the amount of data to be recorded and processed is considerably reduced. We demonstrate this method on human cornea and anterior chamber angle and present, to the best of our knowledge, the first OCT images of retardation and fast axis orientation of the anterior chamber angle region in vivo. PMID- 15128204 TI - Doppler optical coherence imaging of converging flow. AB - The experimental methods of Doppler optical coherence tomography are applied for two-dimensional flow mapping of highly scattering fluid in flow with complex geometry. Converging flow (die entry) is used to demonstrate non-invasive methods to map varying velocity profiles before and after the entry. Complex geometry flow is scanned with approximately 10 x 10 x 10 microm3 spatial resolution. Structural images of the phantom and specific velocity images are demonstrated. A variety of velocity profiles have been obtained before and after the entry. Concave, blunted, parabolic and triangular profiles are obtained at different distances after the entry. Application of the technique to the study of blood circulation is discussed. PMID- 15128205 TI - Correction of distortions in optical coherence tomography imaging of the eye. AB - Optical coherence tomography (OCT) images are affected by artefacts. These artefacts are the result of different factors such as refraction, curvature of the intermediate layers up to the depth of interest and the scanning procedure. The effect of such errors is different, depending on the way the image is acquired, either en-face or longitudinal OCT. We quantify the distortions by evaluating a lateral and an axial error. These measure the lateral and axial deviations of each image point from the object point inside the tissue. We show that the axial distortion can be larger than the achievable depth resolution in modern OCT systems. We have investigated these errors in imaging different tissue: cornea and retina in vivo and an intraocular lens in vitro. PMID- 15128206 TI - The collagen structure of bovine intervertebral disc studied using polarization sensitive optical coherence tomography. AB - Polarization-sensitive optical coherence tomography (PS-OCT) is used to measure the birefringence properties of bovine intervertebral disc and equine flexor tendon. For equine tendon the birefringence delta n is (6.0 +/- 0.2) x 10(-3) at a wavelength of 1.3 microm. This is somewhat larger than the values reported for bovine tendon. The surface region of the annulus fibrosus of a freshly excised intact bovine intervertebral disc displays an identical value of birefringence, delta n = (6.0 +/- 0.6) x 10(-3) at 1.3 microm. The nucleus pulposus does not display birefringence, the measured apparent value of delta n = (0.39 +/- 0.01) x 10(-3) being indistinguishable from the effects of depolarization due to multiple scattering. A clear difference is found between the depth-resolved retardance of equine tendon and that of bovine intervertebral disc. This apparently relates to the lamellar structure of the latter tissue, in which the collagen fibre orientation alternates between successive lamellae. A semi-empirical model based on Jones calculus shows that the measurements are in reasonable agreement with previous optical and x-ray data. These results imply that PS-OCT could be a useful tool to study collagen organization within the intervertebral disc in vitro and possibly in vivo and its variation with applied load and disease. PMID- 15128207 TI - Advanced modelling of optical coherence tomography systems. AB - Analytical and numerical models for describing and understanding the light propagation in samples imaged by optical coherence tomography (OCT) systems are presented. An analytical model for calculating the OCT signal based on the extended Huygens-Fresnel principle valid both for the single and multiple scattering regimes is reviewed. An advanced Monte Carlo model for calculating the OCT signal is also reviewed, and the validity of this model is shown through a mathematical proof based on the extended Huygens-Fresnel principle. Moreover, for the first time the model is verified experimentally. From the analytical model, an algorithm for enhancing OCT images is developed: the so-called true-reflection algorithm in which the OCT signal may be corrected for the attenuation caused by scattering. For the first time, the algorithm is demonstrated by using the Monte Carlo model as a numerical tissue phantom. Such algorithm holds promise for improving OCT imagery and to extend the possibility for functional imaging. PMID- 15128208 TI - Multiple-bandwidth photoacoustic tomography. AB - Photoacoustic tomography, also referred to as optoacoustic tomography, employs short laser pulses to generate ultrasonic waves in biological tissues. The reconstructed images can be characterized by the convolution of the structure of samples, the laser pulse and the impulse response of the ultrasonic transducer used for detection. Although the laser-induced ultrasonic waves cover a wide spectral range, a single transducer can receive only part of the spectrum because of its limited bandwidth. To systematically analyse this problem, we constructed a photoacoustic tomographic system that uses multiple ultrasonic transducers simultaneously, each at a different central frequency. The photoacoustic images associated with the different transducers were compared and analysed. The system was tested by imaging both mouse brains and phantom samples. The vascular vessels in the brain were revealed by all of the transducers, but the image resolutions differed. The higher frequency detectors provided better image resolution while the lower frequency detectors delineated the major structural traits with a higher signal-noise ratio. PMID- 15128209 TI - Fast photoacoustic imaging system based on 320-element linear transducer array. AB - A fast photoacoustic (PA) imaging system, based on a 320-transducer linear array, was developed and tested on a tissue phantom. To reconstruct a test tomographic image, 64 time-domain PA signals were acquired from a tissue phantom with embedded light-absorption targets. A signal acquisition was accomplished by utilizing 11 phase-controlled sub-arrays, each consisting of four transducers. The results show that the system can rapidly map the optical absorption of a tissue phantom and effectively detect the embedded light-absorbing target. By utilizing the multi-element linear transducer array and phase-controlled imaging algorithm, we thus can acquire PA tomography more efficiently, compared to other existing technology and algorithms. The methodology and equipment thus provide a rapid and reliable approach to PA imaging that may have potential applications in noninvasive imaging and clinic diagnosis. PMID- 15128210 TI - Laser speckle imaging of blood flow in microcirculation. AB - Monitoring the spatio-temporal characteristics of microcirculation is crucial for studying the functional activities of biotissue and the mechanism of disease. However, conventional methods used to measure blood flow suffer from limited spatial resolution or the injection of exogenous substances or the need of scanning to obtain the dynamic of regional blood flow. Laser speckle imaging (LSI) technique makes up these disadvantages by obtaining the regional blood flow distribution with high spatio-temporal resolution without the need to scan. In this paper, LSI was introduced to investigate the dynamic responses of the rat mesenteric microcirculation to an incremental dose of phentolamine. The results showed that when the dose of phentolamine was less than 4 microg ml(-1), local application of phentolamine on the mesentery would increase the blood perfusion as the concentration increased. When the dose increased further, the improvement decreased. At a dose of 200 microg ml(-1), a microcirculation impediment was caused. At the same time, different responses between veinules and arterioles were manifested. These suggested that LSI is promising to be a useful contribution to drug development and testing. PMID- 15128211 TI - Hollow-core photonic-crystal fibres for laser dentistry. AB - Hollow-core photonic-crystal fibres (PCFs) for the delivery of high-fluence laser radiation capable of ablating tooth enamel are developed. Sequences of picosecond pulses of 1.06 microm Nd:YAG-laser radiation with a total energy of about 2 mJ are transmitted through a hollow-core photonic-crystal fibre with a core diameter of approximately 14 microm and are focused on a tooth surface in vitro to ablate dental tissue. The hollow-core PCF is shown to support the single-fundamental mode regime for 1.06 microm laser radiation, serving as a spatial filter and allowing the laser beam quality to be substantially improved. The same fibre is used to transmit emission from plasmas produced by laser pulses on the tooth surface in the backward direction for detection and optical diagnostics. PMID- 15128212 TI - The effect of boundary conditions and sample aspect ratio on apparent d33 piezoelectric coefficient determined by direct quasistatic method. AB - The effect of boundary conditions (e.g., type of metal contacts) and sample geometry (e.g., sample aspect ratio) on measurements of direct longitudinal d33 coefficient in piezoelectric ceramics is studied by direct quasistatic method. We show that at small aspect ratio (thickness/lateral dimension <0.1) the measured d33meas is as much as 30% lower than the true value, d33true. Measured d33 increases with increasing aspect ratio and reaches its true value at a threshold aspect ratio that is dependent on ceramic composition and is about approximately 0.5 in the case of soft and hard Pb(Zr,Ti)O3. Experimental results show that, when the force is applied over the whole electroded surface of the sample, the d33 depends only on the aspect ratio and not size of samples. The experimental results are compared with simulations using finite element modeling (FEM) and are interpreted in terms of distribution of strain/stress within the sample, which leads to functional dependence of the measured d33 on transverse d31 and shear d15 coefficients. It is shown experimentally and by FEM that the value of d33 at low aspect ratios depends on the type of metallic contacts used to collect the charge and apply the pressure on the sample. Effect of nonlinearity of the d31 and d15 coefficients on d33 measurements also is considered. PMID- 15128213 TI - Conjugate regenerative dividers. AB - We discuss a novel design of a self-starting regenerative divider that permits division by 3, 4, 5, 6 ... instead of the usual 2. This is accomplished by having the loop oscillate simultaneously at two harmonically related conjugate frequencies, e.g., at nu/4 and 3nu/4. A prototype of the divide-by-four circuit has been constructed for an input frequency of 400 MHz. This divider exhibits very low phase noise, Fourier frequency (1 kHz) = -162 dBc/Hz and Fourier frequency (100 kHz) = -170 dBc/Hz, which is approximately 9 dB lower than that of its constituent parts. Simple modifications of the feedback loop of this circuit enabled it to divide by 3, 5, and 8. Operation at higher division ratios appears feasible under certain conditions. PMID- 15128214 TI - High-frequency, silicon-based ultrasonic nozzles using multiple Fourier horns. AB - This paper presents the design, simulation, and characterization of microfabricated 0.5 MHz, silicon-based, ultrasonic nozzles. Each nozzle is made of a piezoelectric drive section and a silicon resonator consisting of multiple Fourier horns, each with half wavelength design and twice amplitude magnification. Results of finite element three-dimensional (3-D) simulation using a commercial program predicted existence of one resonant frequency of pure longitudinal vibration. Both impedance analysis and measurement of longitudinal vibration confirmed the simulation results with one pure longitudinal vibration mode at the resonant frequency in excellent agreement with the design value. Furthermore, at the resonant frequency, the measured longitudinal vibration amplitude at the nozzle tip increases as the number of Fourier horns (n) increases in good agreement with the theoretical values of 2(n). Using this design, very high vibration amplitude gain at the nozzle tip can be achieved with no reduction in the tip cross-sectional area for contact of liquid to be atomized. Therefore, the required electric drive power should be drastically reduced, decreasing the likelihood of transducer failure in ultrasonic atomization. PMID- 15128215 TI - Investigation of the effects of microbubble shell disruption on population scattering and implications for modeling contrast agent behavior. AB - In a previous study, quantitative measurement of nonlinear scattering revealed some quantitative discrepancies with a model for ultrasound scattering by a population ensemble of microbubbles. This study is designed to investigate the effect of the shell on single-pulse scattering by a population of contrast microbubbles. Nonlinear scattering was measured shortly (15 micros) after exposure to a previous ultrasound pulse with variable intensity. The short time delay eliminated the influence of gas diffusion and focused the study on the consequence of ultrasound exposure. The results suggest that single-pulse scattering is dependent on the disruption properties of the bubble shell and confirm the significance of the shell properties. PMID- 15128216 TI - Acoustic signatures of submicron contrast agents. AB - Previous studies have revealed that hard-shelled submicron contrast agents exhibit large relative expansions and strong acoustical echoes that can be observed experimentally, and predicted by theoretical simulations. In this paper, we study harmonic imaging and pulse-pair imaging techniques designed to assist in the differentiation of these contrast agents from tissue. For harmonic imaging, we apply a high-sensitivity, narrowband strategy that differentiates the microbubble from tissue based on the generation of strong harmonic echoes. For pulse-pair imaging, we apply high spatial resolution, wideband strategies using phase inversion, which relies on the frequency differences observed in response to phase-inverted pulses, and signal subtraction, which takes advantage of the amplitude differences in response to identical pulses. The bubble-to-phantom signal amplitude ratio in the absence of motion approaches 20 dB using phase inversion and 30 dB using signal subtraction; both techniques are robust for up to 50 microm of simulated motion. With the experience gained in these studies, we hope to advance the development of multi-pulse or shaped-pulse techniques that are optimized for specific clinical applications. PMID- 15128217 TI - Singular spectrum analysis applied to backscattered ultrasound signals from in vitro human cancellous bone specimens. AB - Mean scatterer spacing (MSS) holds particular promise for the detection of changes in quasiperiodic tissue microstructures such as may occur during development of disease in the liver, spleen, or bones. Many techniques that may be applied for MSS estimation (temporal and spectral autocorrelation, power spectrum and cepstrum, higher order statistics, and quadratic transformation) characterize signals that contain a mixture of periodic and nonperiodic contributions. In contrast, singular spectrum analysis (SSA), a method usually applied in nonlinear dynamics, first identifies components of signals corresponding to periodic structures and, second, identifies dominant periodicity. Thus, SSA may better separate periodic structures from nonperiodic structures and noise. Using an ultrasound echo simulation model, we previously demonstrated SSA's potential to identify MSS of structures in quasiperiodic scattering media. The current work aims to observe the behavior of MSS estimation by SSA using ultrasound measurements in phantom materials (two parallel, nylon line phantoms and four foam phantoms of different densities). The SSA was able to estimate not only the nylon-line distances but also nylon-line thickness. The method also was sensitive to the average pore-size differences of the four sponges. The algorithms then were applied to characterize human cancellous bone microarchitectures. Using 1-MHz center-frequency, radio-frequency ultrasound signals, MSS was measured in 24 in vitro bone samples and ranged from 1.0 to 1.7 mm. The SSA MSS estimates correlate significantly to MSS measured independently from synchrotron microtomography, r2 = 0.68. Thus, application of SSA to backscattered ultrasound signals seems to be useful for providing information linked to tissue microarchitecture that is not evident from clinical images. PMID- 15128218 TI - Comparison of stress field forming methods for vibro-acoustography. AB - Vibro-acoustography is a method that produces images of the acoustic response of a material to a localized harmonic motion generated by ultrasound radiation force. The low-frequency, oscillatory radiation force (e.g., 10 kHz) is produced by amplitude modulating a single ultrasound beam, or by interfering two beams of slightly different frequencies. Proper beam forming for the stress field of the probing ultrasound is very important because it determines the resolution of the imaging system. Three beam-forming geometries are studied: amplitude modulation, confocal, and x-focal. The amplitude of radiation force on a unit point target is calculated from the ultrasound energy density averaged over a short period of time. The profiles of radiation stress amplitude on the focal plane and on the beam axis are derived. The theory is validated by experiments using a small sphere as a point target. A laser vibrometer is used to measure the velocity of the sphere, which is proportional to the radiation stress exerted on the target as the transducer is scanned over the focal plane or along the beam axis. The measured velocity profiles match the theory. The theory and experimental technique may be useful in future transducer design for vibro-acoustography. PMID- 15128219 TI - Prospects for elasticity reconstruction in the heart. AB - The elastic moduli in anisotropic media can be estimated using either direct mechanical or sound speed measurements. Here we compare moduli in the passive heart estimated with different methods and demonstrate that high-frequency (i.e., ultrasonic) sound speed measurements are inconsistent with static deformations and low-frequency shear wave results. Both tissue fixation and the high-operating frequency of ultrasonic measurements contribute to these discrepancies. Moreover, the precision of ultrasonic sound speed measurements required to estimate elastic moduli describing static deformations of a nearly incompressible anisotropic medium such as the heart appears to be beyond the scope of current methods. We conclude that an incompressible anisotropic elastic model is appropriate for elasticity reconstruction in the heart, in which three independent constants characterize small strain behavior, but four are needed for a fully nonlinear description of finite deformations. PMID- 15128221 TI - Short reflectors operating at the fundamental and second harmonics on 128 degree LiNbO3. AB - In this work, we study numerically the operation of surface acoustic wave (SAW) reflectors comprising a small number of electrodes on the 128 degree YX-cut lithium niobate (LiNbO3) substrate. The electrodes have a finite thickness, and they are either open circuited or grounded. The center-to-center distance between adjacent electrodes d corresponds roughly either to half of the characteristic wavelength d proportional to lambda0/2 or to d proportional to lambda0, for the reflectors operating at the fundamental and second harmonic modes, respectively. We use software based on the finite-element and boundary-element methods (FEM/BEM) for numerical experiments with a tailored test structure having 3 interdigital transducers (IDTs), simulating experimental conditions with an incident wave and reflected and transmitted SAWs. Using the fast Fourier transform (FFT) and time-gating techniques, calculation of the Y-parameters in a wide frequency range with rather a small step allows us to determine the reflection coefficients, and to estimate the energy loss due to bulk-wave scattering. The detailed dependences of the attenuation and reflectivity on the metallization ratio and the electrode thickness are given for the classic 128 degree-cut of LiNbO3. PMID- 15128220 TI - Computed tomography sound velocity reconstruction using incomplete data. AB - An approach based on limited-angle transmission tomography for reconstruction of the sound velocity distribution in the breast is proposed. The imaging setup is similar to that of x-ray mammography. With this setup, the time-of-flight data are acquired by a linear array positioned at the top of the compressed breast that both transmits and receives, and a metal plate is placed at the bottom as a reflector. The setup allows acoustic data acquisition for simultaneous B-mode image formation and the tomographic sound velocity reconstruction. In order to improve the sound velocity estimation accuracy, a new reconstruction algorithm based on a convex programming formulation has been developed. Extensive simulations for both imaging and time-of-flight data based on a 5-MHz linear array were performed on tissues with different geometries and acoustic parameters. Results show that the sound velocity error was generally 1-3 m/s, with a maximum of 5.8 m/s. The radii of the objects under investigation varied from 2 to 6 mm, and all of them were detected successfully. Thus, the proposed approach has been shown to be both feasible and accurate. The approach can be used to complement conventional B-mode imaging to further enhance the detection of breast cancer. PMID- 15128223 TI - Fast FEM/BEM simulation of SAW devices via asymptotic waveform evaluation. AB - The finite element method/boundary element method (FEM/BEM) computation model applied to surface acoustic wave devices requires the solution of a large linear system for each frequency point. An asymptotic waveform evaluation technique is used to obtain an approximate solution of the linear system that is valid over a large frequency bandwidth. The approximate solution was shown to be very accurate and vastly reduces the computation time. PMID- 15128222 TI - Effect of particle bombardment on the orientation and the residual stress of sputtered AlN films for SAW devices. AB - We present a study of the effect of particle bombardment on the preferred orientation and the residual stress of polycrystalline aluminum nitride (AlN) thin films for surface acoustic wave (SAW) applications. Films were deposited on silicon (100) substrates by radio frequency (RF) sputtering of an aluminum target in an argon and nitrogen gas mixture. The main deposition parameters were changed as follows: the total pressure from 4 mTorr to 11 mTorr, the N2 content in the gas mixture from 20% to 80%, and the substrate self-bias voltage from -10 V to 30 V. If a sufficiently high negative substrate self-bias voltage is induced, (00.2)-oriented films are obtained over the full ranges of pressure and N2 content. Such films have values of residual stress ranging from -3 GPa to +1 GPa, depending on the deposition conditions. Our results suggest that the energy of the Ar ions colliding with the substrate controls the preferred orientation of the films, whereas the directionality of the ions (for the same energy) is the main factor determining the residual stress. To demonstrate the suitability of our material for the intended application, SAW filters with good electroacoustic response have been fabricated using AlN thin films with optimized (00.2) orientation and controlled residual stress. PMID- 15128224 TI - Resolution enhancement of experimental echographic images using luminance extrapolation. AB - The experimental results of a resolution enhancement technique are presented, confirming the first simulations previously proposed. The technique enhances the resolution of ultrasound echographic images by extrapolating the luminance changes in the image when the aperture of the transducer is increased, and builds an image that could be obtained with a transducer aperture larger than that physically available. PMID- 15128225 TI - Surface acoustic wave properties of freestanding diamond films. AB - "Ideal" diamond has the highest acoustic velocity of any material known, and is of great interest as a substrate material for high frequency surface acoustic wave (SAW) device structures. However, little is known of the acoustic wave propagation properties of polycrystalline diamond grown by chemical vapour deposition (CVD) techniques, the commercially accessible form of this material. We report on propagation of laser-generated SAW on three forms of freestanding CVD diamond samples, "white" polycrystalline, "black" polycrystalline, and "highly oriented" diamond. Despite differing sample nature, SAW waves propagating along the smooth (nucleation) side of the diamond showed similar velocities in the range 10600-11900 ms(-1). These results are discussed in terms of the potential of each form of CVD diamond for SAW device fabrication. PMID- 15128226 TI - Oxidative C-C bond-forming reaction of electron-rich alkylbenzyl ether with trimethylvinyloxysilane. AB - Treatment of an electron-rich benzyl ether with DDQ at ambient temperature followed by addition of a silyl enol ether undergoes a C-C bond-forming reaction to afford 3-alkoxy-3-phenyl-propionyl compound. This is a general reaction and works well with a variety of silyl enol ethers to give carbonyl products in yields ranging from 10 to 85%. PMID- 15128227 TI - Palladium-catalyzed coupling reaction of terminal alkynes with aryl iodides in the presence of indium tribromide and its application to a one-pot synthesis of 2 phenylindole. AB - The use of a novel PdCl(2)(PPh(3))(2)-InBr(3) reagent system to catalyze cross coupling reactions of a variety of aryl iodides with several terminal alkynes is described. The corresponding functional alkyne derivatives were produced in good to excellent yields. Moreover, a catalytic amount of InBr(3) effectively catalyzes the intramolecular cycloaddition of 2-phenylethynylaniline to form an indole skeleton in high yield. PMID- 15128228 TI - Use of InCl(3) as a cocatalyst and a Cl(2)Pd(DPEphos)-P(2-furyl)(3) catalyst system for one-pot hydrometalation-cross-coupling and carbometalation-cross coupling tandem processes. AB - One-pot conversion of alkynes to regio- and stereodefined alkenylmetals containing Al and Zr via hydrometalation or carbometalation followed by their Pd catalyzed cross-coupling with (E)-ICH=CHBr or (E)-ICH=CHCl proceeds cleanly and selectively to give the corresponding 1-halo-1,3-dienes in excellent yields using a catalyst system consisting of Cl(2)Pd(DPEphos), DIBAL-H, and TFP (catalyst A) with InCl(3) as a cocatalyst. PMID- 15128229 TI - Diastereoselective dearomatization of resorcinols directed by a lactic acid tether: unprecedented enantioselective access to p-quinols. AB - An operationally simple oxidative dearomatization of resorcinol derivatives is reported that employs an inexpensive chiral directing group. The method provides access to a variety of p-quinol derivatives in good yield and diastereoselectivity. A short reductive process affords 4-hydroxy-4 alkylcyclohexenone derivatives in excellent yields and enantiomeric excesses. PMID- 15128230 TI - Bilirubin as an antioxidant: kinetic studies of the reaction of bilirubin with peroxyl radicals in solution, micelles, and lipid bilayers. AB - Bilirubin (BR) showed very weak antioxidant activity in a nonpolar medium of styrene or cumene in chlorobenzene. In contrast, BR exhibited strong antioxidant activity in polar media such as aqueous lipid bilayers or SDS micelles/methyl linoleate (pH 7.4), where the rate with peroxyl radicals, k(inh) = 5.0 x 10(4) M( )(1) s(-)(1), was comparable to that with vitamin E analogues, Trolox, or PMHC. An electron-transfer mechanism accounts for the effect of the medium on the antioxidant properties of BR. PMID- 15128231 TI - Iminium salt catalysts for asymmetric epoxidation: the first high enantioselectivities. AB - A highly enantioselective iminium salt catalyst has been prepared and tested in the catalytic asymmetric epoxidation of unfunctionalized alkenes, giving up to 95% ee, the highest ee yet reported for iminium salt-catalyzed epoxidation. Catalyst loadings as low as 0.1 mol % may be used. PMID- 15128232 TI - Diastereomixture and racemate of myo-inositol derivatives, stronger organogelators than the corresponding homochiral isomers. AB - Contrary to the usually accepted phenomena, an optically heterogeneous 1:1 diastereomixture of DS and LS and a racemate of LS and DR obtained, respectively, from a racemic myo-inositol derivative and (S)- and racemic O-acetylmandelic acid formed stronger organogels, especially with aromatic fluids, than those formed from homochiral isomers, DS and LS. One of the plausible reasons for the formation of the stronger diastereomeric gel is shown to be the complementary interaction of two diastereomers. PMID- 15128233 TI - 2-nitro thioglycoside donors: versatile precursors of beta-D-glycosides of aminosugars. AB - 2-Nitro thioglycosides can be prepared by the Michael addition of thiophenol to 2 nitroglycal derivatives. NIS/TMSOTf activation of these 2-nitro thioglycosides, in the presence of alcohols, rapidly and cleanly led to the desired glycosides in good yield and beta-selectivity. Reduction of the nitro group allowed generation of the corresponding 2-acetamido glycosides. PMID- 15128234 TI - Efficient synthesis of phosphorylated prodrugs with bis(POM)-phosphoryl chloride. AB - An efficient method for the synthesis of phosphorylated prodrugs is described. The preparation of various bis-pivaloyloxymethyl (POM) phosphate triesters was accomplished in moderate to good yields with the use of bis(POM) phosphoryl chloride under mild conditions. PMID- 15128235 TI - Highly selective colorimetric naked-eye Cu(II) detection using an azobenzene chemosensor. AB - Colorimetric azobenzene based chemosensors 1 and 2 were designed for detection of transition-metal ions such as Cu(II) under physiological pH conditions. The internal charge transfer (ICT) sensors are highly colored, absorbing in the green. For 1, the Cu(II) recognition gives rise to red-to-yellow color changes that are visible to the naked-eye and reversible upon addition of EDTA, whereas for 2, which lacks the aromatic o-methoxy chelating group, no such changes were observed. PMID- 15128236 TI - Decomposition of protonated threonine, its stereoisomers, and its homologues in the gas phase: evidence for internal backside displacement. AB - Protonated threonine and its allo diastereomer exhibit different proportions of collisionally activated dissociation (CAD) product ions. N-Methylation attenuates these differences. Water loss from protonated allo-threonine gives protonated trans-3-methylaziridinecarboxylic acid via an internal S(N)2 pathway, rather than protonated vinylglycine. PMID- 15128237 TI - A new simple procedure for discriminating between deracemization and an induced CD Effect in chiral recognition experiments on atropoisomers. AB - A CD band in chiral recognition experiments on racemic stereolabile compounds can be ascribed either to deracemization or to a solely induced CD effect. A procedure is presented that allows one to discriminate positively between the two phenomena. The procedure, based on CD spectroscopy, was used in experiments on racemic biphenylic derivatives in aggregates formed by enantiopure surfactants. In addition to demonstrating a deracemization event, the procedure allowed us to measure the enantiomeric excess. PMID- 15128238 TI - Oligomeric tetrathiafulvalenes: a new route toward conjugated TTF dimers and trimers. AB - The synthesis of bis-TTF 1 and tris-TTF 2, linked by an ethenyl spacer, is described using new useful Wittig-type reagents with the introduction of phosphonate functionality on 1,3-dithiole and TTF frameworks. Electrochemical and spectroscopic studies show intramolecular electronic interactions between conjugated TTF units. PMID- 15128239 TI - Copper(II) acetate promoted oxidative cyclization of arylsulfonyl-o allylanilines. AB - Tosyl-o-allylaniline 1 undergoes oxidative cyclization to produce tetracycle 2 upon treatment with Cu(OAc)(2) and Cs(2)CO(3) at 120 degrees C. The scope of the reaction was extended to other N-sulfonylated aromatic systems. PMID- 15128240 TI - Sterically bulky thioureas as air- and moisture-stable ligands for pd-catalyzed Heck reactions of aryl halides. AB - We demonstrate that sterically bulky N,N'-disubstituted cyclic thiourea-Pd(0) complexes are air- and moisture-stable and highly active catalysts for palladium catalyzed Heck reaction of aryl iodides and bromides with olefins (TONs up to 500000 for the reaction of PhI and methyl acrylate). Even activated aryl chlorides can undergo complete conversion in Bu(4)NBr in the presence of 1 mol % Pd catalyst. PMID- 15128241 TI - Efficient syntheses of heterocycles and carbocycles by electrophilic cyclization of acetylenic aldehydes and ketones. AB - Highly substituted oxygen heterocycles can be prepared in good yields at room temperature by reacting o-(1-alkynyl)-substituted arene carbonyl compounds with NBS, I(2), ICl, p-O(2)NC(6)H(4)SCl, or PhSeBr and various alcohols or carbon based nucleophiles. Naphthyl ketones and iodides are readily prepared by the reaction of 2-(1-alkynyl)arene-carboxaldehydes with I(2) and simple olefins or alkynes. PMID- 15128242 TI - Synthesis of aminoshikimic acid. AB - 5-amino-5-deoxyshikimic acid (aminoshikimic acid) was synthesized from glucose using recombinant Amycolatopsis mediterranei and also synthesized by a tandem, two-microbe route employing Bacillus pumilus and recombinant Escherichia coli. PMID- 15128243 TI - Formation of 3-halobenzyne: solvent effects and cycloaddition adducts. AB - Noncoordinating solvents permit the halogen-metal exchange-induced formation of benzyne (aryne) from di- and trihalobenzene precursors in the presence of cyclopentadiene to give 1,4-dihydro-1,4-methano-naphthalenes. Studies with mixed halide precursors and nonacidic Diels-Alder diene traps reveal that ethereal and hydrocarbon solvents influence the halide leaving group facility, resulting in a reversal of 3-halobenzyne regioselectivity. PMID- 15128245 TI - Highly diastereoselective epoxidation of allyl-substituted cycloalkenes catalyzed by metalloporphyrins. AB - Highly diastereoselective epoxidations of allyl-substituted cycloalkenes including allylic alcohols, esters, and amines using sterically bulky metalloporphyrins [Mn(TDCPP)Cl] (1) and [Ru(TDCPP)CO] (2) as catalysts have been achieved. The "1 + H(2)O(2)" and "2 + 2,6-Cl(2)pyNO" protocols afforded trans epoxides selectively in good yields (up to 99%) with up to >99:1 trans selectivity. PMID- 15128244 TI - Coriatone and corianlactone, two novel sesquiterpenes from Coriaria nepalensis. AB - Both coriatone (1). a novel highly oxygenated picrotoxane-type sesquiterpene, and corianlactone (2). with an unprecedented sesquiterpene basic skeleton, named coriane, were isolated from Coriaria nepalensis Wall. The structures of 1 and 2 were determined by analysis of their two-dimensional NMR data, and the structure of 2 was confirmed by X-ray analysis. Compounds 1 and 2 showed no remarkable inhibitory activity toward K(562) cells. They are cytotoxic with IC(50) > 50 microg/mL (cis-platinim: IC(50) = 0.49 microg/mL). PMID- 15128246 TI - Stereoselective synthesis of d-desosamine and related glycals via tungsten catalyzed alkynol cycloisomerization. AB - Stereoselective synthesis of d-desosamine diacetate ester (iii, R = Ac) was achieved from the glycal (ii). generated by tungsten carbonyl-catalyzed cycloisomerization of the corresponding amino-alkynol (i). A wide variety of N substituents (R, R') are compatible with the cycloisomerization, provided that at least one R or R' is an acyl derivative. PMID- 15128247 TI - Polyaryl anion radicals via alkali metal reduction of arylurea oligomers. AB - A series of N-methylated polyarylurea oligomers have been reduced with potassium metal in HMPA. These reductions result in the transient formation of arylurea anion radicals, which undergo reductive elimination of the urea linkages. The aryl moieties appear in the products as the anion radicals of oligoaryl systems. The reaction is intramolecular, and the sequencing in the polyaryl anion radical remains the same as in the polyarylureas due to the urea-enforced pi-pi stacking interactions. PMID- 15128249 TI - The asymmetric power of chiral ligands determined by competitive asymmetric autocatalysis. AB - Asymmetric autocatalytic reactions were initiated by using two competing chiral ligands bearing opposite configurations. The absolute configuration of the resulting highly enantioenriched product reflects the different efficiencies of the two catalysts. Thus, our method provides a simple and efficient way to compare the asymmetric power of chiral ligands for enantioselective catalysis both qualitatively and quantitatively. PMID- 15128248 TI - Robust, soluble pentacene ethers. AB - We report the synthesis and characterization of a series of alkoxy-substituted silylethynylated pentacene derivatives (R = CH(2)CH(2), CHCH, CH(2)). All three compounds are easily prepared, soluble in common organic solvents, and stable both as solids and in solution. Two of the derivatives possess significant pi face interactions in the crystal. Values for lambda(max) for these new pentacene derivatives range from 621 to 674 nm, and oxidation potentials lie between 109 and 301 mV versus ferrocene. PMID- 15128250 TI - Thiabowls: synthesis, molecular structure, and novel supramolecular architecture of trithia-[3]-peristylane. AB - A synthesis of trithia-[3]-peristylane, C(6)H(6)S(3), a novel C(3)(v)() symmetric thiabowl, from the exotic hydrocarbon bullvalene has been accomplished. The X-ray crystal structure of this trithiabowl displays an unprecedented supramolecular architecture in the solid state with 12 CH.S interactions involving all of its six hydrogen atoms and the two lone pairs on each of the three sulfur atoms. PMID- 15128251 TI - Highly selective intra- and intermolecular coupling reactions of diazo compounds to form cis-alkenes using a ruthenium porphyrin catalyst. AB - [Ru(2,6-Cl(2)TPP)(CO)] catalyzed intramolecular coupling reactions of bisdiazoacetates and intermolecular coupling reactions of monodiazoacetates to afford the coupling products in up to 76% and 93% yields, respectively. Only the cis isomers were obtained from the reactions. Employing such a ruthenium catalyzed coupling reaction of a diazo compound as a key step allowed the synthesis of Patulolide B in 67% yield with a ratio of >40:1 against its trans isomer. PMID- 15128252 TI - Highly regioselective ring-opening/cross-metathesis reactions of 2 sulfonylnorbornene derivatives. AB - Tandem ring-opening/cross-metathesis (ROM/CM) reactions of norbornenes can be a powerful entry into highly substituted organic molecules. However, their utility has been limited largely to symmetrical norbornenes because of the general lack of regioselective variants of these reactions. This manuscript describes our successful attempts to address this issue through the use of a sulfone to direct the ROM/CM reaction of a 7-azanorbornene and a 7-oxanorbornene. PMID- 15128253 TI - Solid-phase synthesis of constrained terminal and internal lactam peptidomimetics. AB - Lactams are key components of many peptidomimetic structures. Five- and six membered lactam peptidomimetics with hydrogen or amino acid side chains at the alpha-position can be constructed from peptide precursors during a solid-phase synthesis. There is no significant racemization of remote stereocenters during synthesis. PMID- 15128255 TI - Enantioselective synthesis of allylic alcohols by the sequential aminoxylation olefination reactions of aldehydes under ambient conditions. AB - A novel, highly enantioselective synthesis of O-amino-substituted allylic alcohols by the sequential asymmetric alpha-aminoxylation/Wadsworth-Emmons-Horner olefination reactions of aldehydes is presented. PMID- 15128254 TI - Polychlorinated androstanes from the burrowing sponge Cliona nigricans. AB - Two new steroidal derivatives, named clionastatins A and B, have been isolated from the burrowing sponge Cliona nigricans. These molecules are tri-and tetrachlorinated androstane derivatives, respectively, and they represent the first polyhalogenated steroids found in a natural organism, either marine or terrestrial, and the first examples of halogenated androstanes in nature. Both clionastatins proved to be potently cytotoxic. PMID- 15128256 TI - Distinguishing between abstraction and addition as the first step in the reaction of a nitroxyl radical with cyclohexene. AB - An unambiguous method for distinguishing between abstraction-addition and addition-abstraction mechanisms (and mixtures thereof) in the reaction of 4 hydroxy-2,2,6,6-tetramethylpiperidine-N-oxyl with a specifically deuterated cyclohexene, 1,2-dideuteriocyclohexene, is demonstrated. PMID- 15128257 TI - Highly stereoselective addition of organometallic reagents to N-tert butanesulfinyl imines derived from 3- and 4-substituted cyclohexanones. AB - Addition of alkyl or aryl Grignard reagents to N-sulfinyl imines derived from 3- and 4-substituted cyclohexanones proceeds with good yields and with excellent diasteroselectivity. The selectivity of the reaction is controlled by the ring substituent rather than the sulfinyl group stereochemistry, and therefore racemic tert-butanesulfinamide can be employed. PMID- 15128259 TI - 2-tert-butyl-3-methyl-2,3-dihydroimidazol- 4-one-N-oxide: a new nitrone-based chiral glycine equivalent. AB - Cycloaddition reactions between a new homochiral imidazolone-derived nitrone afford cycloadducts in high yield and with high stereoselectivity. Subsequent cycloadduct elaboration affords the gamma-lactones of gamma-hydroxy-alpha-amino acids as well as the optically pure amino acids themselves. PMID- 15128258 TI - With regard to the hydrogen bonding in complexes of pyridylureas, less is more. a role for shape complementarity and CH...O interactions? AB - A pyridylurea.tetraazaanthracenedione complex with three hydrogen bonds is more stable than an analogous complex with four hydrogen bonds. An X-ray analysis and modeling suggests a steric mismatch destabilizing the latter and a CH...O contact enhancing the stability of the former. PMID- 15128260 TI - Catalyst and ring size effects on periselectivity of oxonium ylide rearrangements. AB - Diazoketones were subjected to carbene-transfer with Rh(II) or Cu(II) catalysts to probe the selectivity for rearrangement via five- or six-membered oxonium ylides. 4,5-Bis(benzyloxy) and 4-allyloxy-5-benzyloxy substrates 3a,b showed a large preference for rearrangement via the five-membered ylide under all conditions. However, a sharp divergence was seen with 5-allyloxy-4-benzyloxy substrate 3c, which underwent predominantly a [2,3]-shift to pyran 5c via the six membered ylide with Cu(II) catalysis and a [1,2]-shift to furan 4c via the five membered ylide with Rh(II) catalysis. PMID- 15128261 TI - Bielschowskysin, a gorgonian-derived biologically active diterpene with an unprecedented carbon skeleton. AB - Bielschowskysin is a naturally occurring diterpene isolated from the Caribbean gorgonian octocoral Pseudopterogorgia kallos. Its highly oxygenated hexacyclic structure is based on a previously undescribed tricyclo[9.3.0.0(2,10)]tetradecane ring system that was established through spectroscopic analysis and X-ray crystallographic analysis. Bielschowskysin was shown to exhibit antimalarial activity against Plasmodium falciparum as well as strong anticancer activity against two human cancer cell lines. PMID- 15128262 TI - Selective functionalization in positions 2 and 3 of indole via an iodine-copper exchange reaction. AB - Two selectively successive I/Cu exchange reactions performed with (Nphyl)CuLi allow the functionalization of indoles in positions 2 and 3. The 2,3-diketones prepared by this method can readily be converted to tricyclic heterocycles by standard methods. PMID- 15128263 TI - Novel selectfluor and deoxo-fluor-mediated rearrangements. New 5(6)-methyl and phenyl methanopyrrolidine alcohols and fluorides. AB - Stereoselective syntheses of novel 5,6-difunctionalized-2 azabicyclo[2.1.1]hexanes containing 5-anti-fluoro or hydroxyl in one methano bridge and a variety of syn- or anti-chloro, fluoro, hydroxy, methyl, or phenyl substituents in the other methano bridge have been effected. Rearrangements of iodides to alcohols were initiated using Selectfluor. Rearrangement of alcohols to fluorides was initiated using Deoxo-Fluor. Ring opening of 2 azabicyclo[2.2.0]hex-5-ene exo-epoxide with organocopper reagents is regioselective at C(5). PMID- 15128264 TI - Solid-phase synthesis of anandamide analogues. AB - The endocannabinoids are amides and esters of arachidonic acid that can mimic the pharmacological properties of Delta(9)-tetrahydrocannabinol (Delta(9)-THC). Anandamide, the most prominent of the endocannabinoids, has been implicated in both metabolic/physiological roles of the central nervous system, making it an attractive medicinal target. As such, we report the first solid-phase methodology that expedites access to various anandamide analogues. Our synthesis features a repetitive Cu-mediated coupling reaction between terminal alkynes and propargyl halides or allylic halides. PMID- 15128265 TI - Diastereoselectivity-switchable and highly enantioselective 1,3-dipolar cycloaddition of nitrones to alkylidene malonates. AB - Trisoxazoline 1/Co(ClO(4))(2).6H(2)O catalyzed the 1,3-cycloaddition between nitrones 3 with alkylidene malonates 2 at 0 degrees C to give the isoxazolidines with both high enantioselectivity and high exo selectivity. However, when the temperature was lowered from 0 to -40 degrees C, the same cycloaddition afforded endo isomers as the major products with good to high enantioselectivity. A mechanism is provided. PMID- 15128266 TI - Highly efficient enantiospecific synthesis of imidazoline-containing amino acids using bis(triphenyl)oxodiphosphonium trifluoromethanesulfonate. AB - A highly efficient enantiospecific synthesis of imidazoline-based amino acids is reported from dipeptides composed of a C-terminal beta-amino-alpha-amino acid residue using bis(triphenyl) oxodiphosphonium trifluoromethanesulfonate. These imidazolines were easily converted to imidazoles and incorporated into macrolactam analogues of bistratamide H without loss of stereochemical integrity. PMID- 15128267 TI - Rhodium-catalyzed direct C-H addition of 4,4-dimethyl-2-oxazoline to alkenes. AB - A new method for the preparation of 2-substituted oxazolines by rhodium-catalyzed coupling of alkenes with 4,4-dimethyl-2-oxazoline is reported. The oxazoline products are obtained in good yield with excellent selectivity for the linear product. A variety of alkene substitution patterns and functional groups are tolerated. This procedure represents an attractive alternative to hydroesterification because it does not involve the manipulation of CO gas. PMID- 15128268 TI - Cellular effects of deoxynojirimycin analogues: uptake, retention and inhibition of glycosphingolipid biosynthesis. AB - Deoxynojirimycin (DNJ) analogues are inhibitors of ceramide glucosyltransferase (CGT), which catalyses the first step in the glucosphingolipid (GSL) biosynthetic pathway. We have synthesized a series of DNJ analogues to study the contribution of N-alk(en)yl side chains (C4, C9 or C18) to the behaviour of these analogues in cultured HL60 cells. When cells were treated for 16 h at non-cytotoxic concentrations of inhibitor, a 40-50% decrease in GSL levels was measured by HPLC analysis of GSL-derived oligosaccharides following ceramide glycanase digestion of GSL and 2-aminobenzamide labelling of the released oligosaccharides. Using a novel technique for short-term [14C]galactose labelling of cellular GSL, we used compound inhibition of GSL biosynthesis as a marker for compound uptake into cells. Surprisingly, the uptake of all three of the DNJ analogues was extremely rapid and was not dependent upon the length of the N-alk(en)yl moiety. Compound uptake occurred in less than 1 min, as shown by the complete inhibition of GSL labelling in cells treated with all the DNJ analogues. Greatly increased cellular retention of N-cis-13-octadecenyl-DNJ was observed relative to the shorter-chain compounds, N-butyl-DNJ and N-nonyl-DNJ, as indicated by complete inhibition of CGT 24 h after removal of inhibitor from the culture medium. The present study further characterizes the properties of N-alk(en)ylated DNJs, and demonstrates that increasing the length of the side chain is a simple way of improving imino sugar retention and therefore inhibitory efficacy for CGT in cultured cells. PMID- 15128270 TI - Role of the hypothalamic-pituitary-adrenal axis, glucocorticoids and glucocorticoid receptors in toxic sequelae of exposure to bacterial and viral products. AB - The hypothalamic-pituitary-adrenal (HPA) axis is activated during many bacterial and viral infections, resulting in an increase in circulating glucocorticoid levels. This HPA axis activation and glucocorticoid response are critical for the survival of the host, as demonstrated by the fact that removal of the HPA axis (by adrenalectomy or hypophysectomy) or glucocorticoid receptor (GR) blockade enhances the severity of the infection and in some cases enhances the mortality rate. Replacement with a synthetic glucocorticoid reverses these effects by reducing the severity of the infection and provides protection against lethal effects. In addition, some bacteria and viral infections have been shown to affect the GR directly. These have been described and the implications of such an effect discussed. PMID- 15128269 TI - Targeting of Helicobacter pylori vacuolating toxin to lipid raft membrane domains analysed by atomic force microscopy. AB - The Helicobacter pylori vacuolating toxin VacA causes several effects on mammalian cells in vitro, including intracellular vacuolation, formation of pores in the plasma membrane and apoptosis. When added to cells, VacA becomes associated with detergent-resistant membranes, indicating that it binds preferentially to lipid rafts. In the present study, we have used atomic force microscopy to examine directly the association of VacA with lipid domains in supported lipid bilayers. VacA did not bind to lipid bilayers at pH 7.6. In contrast, at pH 4.0, VacA associated with the bilayers in the form of 26-nm oligomeric complexes. VacA bound to bilayers produced from either brain lipids or SM (sphingomyelin) plus cholesterol, each of which lacked detectable lipid domains. Bilayers composed of DOPC (dioleoylphosphatidylcholine), SM and cholesterol contained clearly visible raft-like domains, and VacA preferentially associated with these rafts. VacA bound poorly to raft-like domains in DOPC/SM bilayers, indicating that cholesterol is required for efficient association of VacA with lipid domains. When PS (phosphatidylserine), an anionic phospholipid that does not partition significantly into rafts, was added to the mixture of DOPC, SM and cholesterol, VacA was excluded from the rafts, indicating that it binds more avidly to PS than to the raft components. A typical plasma membrane exhibits pronounced lipid asymmetry, with SM enriched in the outer leaflet and PS in the inner leaflet. Therefore it is probable that the association of VacA with rafts in DOPC/SM/cholesterol bilayers represents a useful model for understanding the interactions of VacA with membranes in vivo. PMID- 15128271 TI - Testosterone-dependent variations in plasma and intrapituitary corticosteroid binding globulin and stress hypothalamic-pituitary-adrenal activity in the male rat. AB - Hypothalamic-pituitary-adrenal (HPA) activity is governed by glucocorticoid negative feedback and the magnitude of this signal is determined, in part, by variations in plasma corticosteroid-binding globulin (CBG) capacity. Here, in gonadectomized male rats we examine the extent to which different testosterone replacement levels impact on CBG and HPA function. Compared with gonadectomized rats with low testosterone replacement ( approximately 2 ng/ml), plasma adrenocorticotropin and beta-endorphin/beta-lipotropin responses to restraint stress were reduced in gonadectomized rats with high testosterone replacement ( approximately 5 ng/ml). Plasma CBG levels also varied negatively as a function of testosterone concentration. Moreover, glucocorticoid receptor binding in the liver was elevated by higher testosterone replacement, suggesting that testosterone acts to enhance glucocorticoid suppression of CBG synthesis. Since pituitary intracellular CBG (or transcortin) is derived from plasma, this prompted us to examine whether transcortin binding was similarly responsive to different testosterone replacement levels. Transcortin binding was lower in gonadectomized rats with high plasma testosterone replacement ( approximately 7 ng/ml) than in gonadectomized rats with low testosterone replacement ( approximately 2 ng/ml). This testosterone-dependent decrease in pituitary transcortin was associated, in vitro, with an enhanced nuclear uptake of corticosterone. These findings indicate that the inhibitory effects of testosterone on corticotrope responses to stress may be linked to decrements in plasma and intrapituitary CBG. This could permit greater access of corticosterone to its receptors and enhance glucocorticoid feedback regulation of ACTH release and/or proopiomelanocortin processing. PMID- 15128272 TI - Possible expression of functional glutamate transporters in the rat testis. AB - Neither expression nor functionality is clear in peripheral tissues with the molecular machineries required for excitatory neurotransmitter signaling by L glutamate (Glu) in the central nervous system, while a recent study has shown that several Glu receptors are functionally expressed in the rat testis. This fact prompted us to explore the possible functional expression in the rat testis of the Glu transporters usually responsible for the regulation of extracellular Glu concentrations in the brain. RT-PCR revealed the expression, in the rat testis, of mRNA for five different subtypes of Glu transporters, in addition to that for particular subtypes of ionotropic and metabotropic Glu receptors. Glutamate transporter-1 (GLT-1) was different in the brain from that in the testis in terms of molecular sizes on Northern and Western blot analyses. In situ hybridization as well as immunohistochemical analysis showed localized expression of glutamate aspartate transporter at interstitial spaces and GLT-1 at elongated spermatids in the rat testis respectively. The expression of mRNA was localized for excitatory amino acid transporter-5 at the basal compartment of the seminiferous tubule in the rat testis. [(3)H]Glu was accumulated in testicular crude mitochondrial fractions in a temperature- and sodium-dependent saturable manner with pharmacological profiles similar to those shown in brain crude mitochondrial fractions. These results suggested that particular subtypes of central Glu transporters for the regulation of extracellular Glu concentrations in the rat testis could be constitutively and functionally expressed. PMID- 15128273 TI - Effects of PNU157706, a dual 5alpha-reductase inhibitor, on gene expression in the rat epididymis. AB - The epididymis is the site of sperm maturation and storage. 5alpha-Reductases (types 1 and 2) are key enzymes in this tissue because they convert testosterone to dihydrotestosterone (DHT), the main androgen regulating epididymal functions. Examining the consequences of inhibiting DHT formation is likely to provide important information regarding the regulation of epididymal functions, yet few inhibitor studies have focused on this tissue. To understand better DHT-mediated regulation of epididymal gene expression, we employed a dual 5alpha-reductase inhibitor and cDNA microarrays to examine the effects of 5alpha-reductase inhibition on gene expression in the initial segment, caput, corpus, and cauda epididymidis. Inhibition of epididymal 5alpha-reductase activity by PNU157706 was confirmed by in vitro enzyme assays. Rats were treated with 0, 0.1, 1.0 or 10 mg/kg per day PNU157706 for 28 days. The weights of DHT-dependent tissues, including the epididymis, were decreased following treatment. The effect of treatment on gene expression was dose-dependent and highly segment-specific. The initial segment responded uniquely in that a similar number of genes increased and decreased in expression compared with the other segments where the majority of affected genes decreased in expression. Some of the more dramatically affected genes were involved in signal transduction as well as fatty acid and lipid metabolism, regulation of ion and fluid transport, luminal acidification, oxidative defense and protein processing and degradation. These are essential processes contributing to the formation of an optimal luminal microenvironment required for proper sperm maturation. These results provide a novel insight into the DHT-dependent mechanisms that control epididymal functions. PMID- 15128275 TI - GH and insulin affect fatty acid synthase activity in isolated porcine adipocytes in culture without any modifications of sterol regulatory element binding protein 1 expression. AB - The ability of GH to decrease fatness and insulin-regulated events such as lipogenic enzyme activities is well known in pigs. Nevertheless, the precise mechanism underlying these actions has not been elucidated yet. Expression of the transcription factor sterol regulatory element binding protein (SREBP)-1 has been reported as a key mediator of insulin action in rat hepatocytes and adipose cell lines. The present study aimed to determine whether the regulation of lipogenesis by GH and/or insulin in porcine adipocytes also involved SREBP-1. Isolated adipocytes, obtained from perirenal or s.c. adipose tissue samples of female pigs (51+/-0.4 kg; n=17), were cultured in serum-free medium in the absence or presence of these hormones for up to 4 days. Glucose incorporation and fatty acid synthase activity were increased by insulin in a dose-dependent manner in adipocytes of both sites. The increase was maximal at 1.7 and 17 nM in s.c. and perirenal adipocytes respectively, suggesting inter-depot differences in the regulation of lipogenesis by insulin. These insulin-stimulated events were decreased by GH (1 nM). No change in SREBP-1 mRNA levels was observed in response to GH and/or insulin. Taken together, these data indicate that the regulation of lipogenesis by insulin and GH appears to not involve changes in SREBP-1 mRNA levels in porcine adipocytes. PMID- 15128274 TI - Aurora B expression in normal testis and seminomas. AB - Aurora/Ipl1-related kinases are a conserved family of proteins that have multiple functions during mitotic progression. High levels of Aurora kinases are characteristic of rapidly dividing cells and tumours. Aurora B encodes a protein that associates with condensing chromatin, concentrates at centromeres, and then relocates onto the central spindle at anaphase. In this study the expression and the localisation of Aurora B throughout germinal epithelial progression in normal testis and its neoplastic counterpart were analysed. Immunocytochemistry and RT PCR analysis of mouse germinal epithelium cells showed the presence of Aurora B in spermatogonia and occasionally in spermatocytes. Western blot analysis revealed the typical Aurora B isoform ( approximately 41 kDa) in the same cellular types. A similar distribution was observed in human testis by immunohistochemistry. Moreover, the distribution and the expression of Aurora B were investigated in neoplasms derived from germ cells. Surgical samples of seminomas were analysed, and a high percentage of Aurora B positive cells (51%) was detected; the expression of Aurora B was significantly related to the MIB-1 proliferation marker (R=0.816). The data presented here demonstrate that Aurora B expression occurs in spermatogonial division. Furthermore, our results indicate that the expression of Aurora B is a consistent feature of human seminomas. PMID- 15128276 TI - Dairy cows experience selective reduction of the hepatic growth hormone receptor during the periparturient period. AB - At parturition, dairy cows experience a 70% reduction in plasma IGF-I. This reduction coincides with decreased abundance of GHR1A, the liver-specific transcript of the growth hormone receptor (GHR) gene, suggesting impaired growth hormone-dependent synthesis of IGF-I. It is not immediately obvious that the periparturient reduction in GHR1A is sufficient to reduce hepatic GHR abundance. This is because approximately 50% of total GHR mRNA abundance in prepartum liver is accounted for by ubiquitously expressed transcripts which remain collectively unchanged at parturition. In addition, the possibility that parturition alters GHR expression in other growth hormone target tissue has not been examined. To address these questions, we measured GHR gene expression and GHR protein in liver and skeletal muscle of four dairy cows on days -35,+3 and+56 (relative to parturition on day 0). Hepatic GHR abundance and GHR1A transcripts were lower on day+3 than on day -35 and returned to late pregnancy value by day+56. Additional studies in two other groups of cows indicated that the hepatic levels of the GHR protein recovered substantially within 10 days after parturition. These changes occurred without variation in the abundance of HNF4, a liver-enriched transcription factor activating the promoter responsible for GHR1A synthesis. In contrast to liver, levels of GHR gene expression and GHR protein were identical on days -35,+3 and+56 in skeletal muscle. These data suggest a role for the GHR in regulating tissue-specific changes in growth hormone responsiveness in periparturient dairy cows. PMID- 15128277 TI - Prenatal stress induces intrauterine growth restriction and programmes glucose intolerance and feeding behaviour disturbances in the aged rat. AB - There is growing evidence that prenatal adversities could be implicated in foetal programming of adult chronic diseases. Since maternal stress is known to disturb the foetal glucocorticoid environment, we examined the consequences of prenatal stress on foetal growth, on glucose-insulin metabolism and on feeding behaviour in the aged male rat. In foetuses at term, maternal stress reduced body, adrenal and pancreas weight as well as plasma corticosterone and glucose levels. In aged male rats (24 months of age), prenatal stress induced hyperglycaemia and glucose intolerance and decreased basal leptin levels. Moreover, after a fasting period, they showed an increased food intake. These data suggest that maternal stress induces a long-lasting disturbance in feeding behaviour and dysfunctions related to type 2 diabetes mellitus. This programming could be linked to the early restricted foetal growth and to the adverse glucocorticoid environment in utero. PMID- 15128278 TI - Caloric restriction reverses the deficits in leptin receptor protein and leptin signaling capacity associated with diet-induced obesity: role of leptin in the regulation of hypothalamic long-form leptin receptor expression. AB - The objectives of this study were to determine if reduced long-form leptin receptor (ObRb) expression in diet-induced obese (DIO) animals is associated with deficits in maximal leptin signaling and, secondly, to establish the effects of short-term caloric restriction (CR) on ObRb expression and function. Groups of DIO and life-long chow-fed (CHOW) F344xBN male rats, aged 6 months, were given an i.c.v. injection containing 2 micro g leptin or artificial cerebrospinal fluid (ACSF) vehicle. Leptin induced a >6-fold increase in STAT3 phosphorylation in CHOW rats, but less than 2-fold increase in DIO. Reduced maximal leptin stimulated STAT3 phosphorylation in DIO rats was coupled with a decline in both ObRb expression and protein. At this point, subgroups of DIO and CHOW animals underwent CR for 30 days and were then tested for acute leptin responsiveness. CR resulted in a 45 and 85% increase respectively in leptin-stimulated STAT3 phosphorylation in CHOW and DIO animals. Similarly, CR increased ObRb expression and protein in both CHOW and DIO animals. To explore the role of leptin in regulating ObRb expression, we reversibly overexpressed leptin in the hypothalamus and found that ObRb mRNA inversely follows central leptin expression. By enhancing both ObRb expression and signaling capacity, CR may enhance leptin responsiveness in leptin-resistant DIO animals. PMID- 15128279 TI - Effect of heparin administration to sheep on the release profiles of circulating activin A and follistatin. AB - Activin A and follistatin are normally present in relatively low amounts in the circulation. Heparin administration elicits a rapid and robust release of these proteins, although this phenomenon is poorly defined. In the present studies, the response to heparin administration was evaluated in the plasma of adult ewes in terms of whether it was dose-dependent, could be neutralized, was responsive to multiple stimulation, and the nature of the activin A and follistatin released. Activin A and follistatin were rapidly released by heparin in a dose-dependent manner (25, 100 or 250 IU/kg), with differences in the response as adjudged by peak concentration, timing of the peak and area under the curve. The heparin response could be blocked by pretreatment with protamine; conversely protamine injection alone (2 mg/kg) elicited release of follistatin but not activin A. Repeat administration of heparin at three-hourly intervals resulted in activin and follistatin responses to each injection, but each subsequent stimulation increased and extended the responses, consistent with saturation of the heparin clearance mechanism. Size exclusion chromatography of plasma samples confirmed that the majority of activin and follistatin released by heparin was a complex, whereas follistatin released by protamine was unbound. These data are consistent with a large pool of activin A and follistatin resident on extracellular matrices, with the rapid response implicating the vascular endothelium as the prime site of release following administration of these commonly used anticoagulant therapies. PMID- 15128280 TI - Divergent roles for ferric ions in the biological activity of amidated and non amidated gastrins. AB - Amidated forms of the peptide hormone gastrin act via the cholecystokinin-2 receptor to stimulate gastric acid secretion, whereas non-amidated forms stimulate colonic mucosal proliferation via a novel, as yet uncharacterised, receptor. Nuclear magnetic resonance (NMR) and fluorescence spectroscopic studies have revealed that glycine-extended gastrin17 bound two ferric ions, and that ferric ion binding was essential for biological activity. We have therefore investigated the role of ferric ions in the biological activity of amidated gastrin17. As with glycine-extended gastrin17, fluorescence quenching experiments indicated that Glu7 Ala and Glu8,9 Ala mutants of amidated gastrin17 each bound only one ferric ion. The affinity of the mutant peptides for the cholecystokinin 2 receptor on transfected COS-7 cells or on Tlymphoblastoid Jurkat cells, and their potency in stimulation of proliferation in Jurkat cells and inositol phosphate production in transfected COS-7 cells, were similar to the values obtained for amidated gastrin17. In addition, the iron chelator desferrioxamine did not significantly inhibit either binding of amidated gastrin17 to the cholecystokinin-2 receptor, or stimulation of inositol phosphate production by amidated gastrin17 in transfected COS-7 cells. We conclude that, in contrast to glycine-extended gastrin17, binding of ferric ions is not essential for the biological activity of amidated gastrin17. Our results support the concept of distinct modes of action for amidated and non-amidated gastrins, and raise the possibility of developing selective antagonists of the actions of non-amidated and amidated gastrins. PMID- 15128281 TI - Targets of 17beta-oestradiol-induced apoptosis in colon cancer cells: a mechanism for the protective effects of hormone replacement therapy? AB - Epidemiological studies show a strong link between postmenopausal hormone replacement therapy and decreased incidence of colorectal cancer. The colon cancer cell line, COLO 205, develops sensitivity to 17beta-oestradiol (E(2)) in apoptosis assays with increasing passage number (>40), and we hypothesised that genes selectively regulated in multiply passaged cells were likely to be important in E(2)-related apoptosis. Gene array analysis was used to compare the patterns of genes up- or down-regulated in E(2)-sensitive and -insensitive cells. For some genes, changes in mRNA expression were confirmed by protein expression analyses. Changes found in response to E(2) in multiply passaged cells, but not minimally passaged cells, included induction of growth arrest and DNA damage inducible protein 153 (GADD153), and repression of Kirsten-Ras 2B (K-Ras-2B), metastasis inhibition factor NM23 and vascular endothelial growth factor. A second group of genes was regulated with E(2) exposure in both cell types, and is unlikely to be critically involved in E(2)-associated apoptosis. These included up-regulation of butyrate response factor 1 (BRF1) and down-regulation of c-jun and the breast cancer associated ring domain gene known as BARD1. By comparing control arrays from the two cell populations, cAMP-response element-binding protein (CBP), which is associated with steroid receptor-dependent target gene transcription and the oncoprotein, tyrosine kinase-T3 (TRK-T3), were up-regulated whereas retinoic acid receptor alpha (RARalpha) was down-regulated in multiply passaged cells. This study provides evidence for selective regulation of genes in colon cancer cells by E(2), indicates which of those regulated are likely to be involved in induced apoptosis, and suggests genes likely to be responsible for facilitation. PMID- 15128282 TI - Increased adrenomedullin expression in lungs in endotoxaemia. AB - Adrenomedullin (AM) is a peptide involved in cardiovascular homeostasis and in inflammation. We examined its expression in a rat model of endotoxaemia. Male Sprague-Dawley rats received intraperitoneal injection of 5 or 10 mg/kg lipopolysaccharide (LPS), or saline as control. Rats were killed at 1, 3, 6, 12 and 24 h after injection. LPS at 5 mg/kg, but not saline, increased plasma AM significantly at 3 h. At 10 mg/kg, plasma AM was raised at 3, 6 and 12 h. Immunoreactive AM concentration in lung increased after 5 or 10 mg/kg LPS, but not saline. PreproAM mRNA level in lung was significantly increased at 3 and 6 h. In conclusion, endotoxin stimulates the expression of AM in the lungs and increases its circulatory concentration. AM may be involved in the systemic response to sepsis. PMID- 15128283 TI - Brevinin-1 and multiple insulin-releasing peptides in the skin of the frog Rana palustris. AB - Few studies have comprehensively examined amphibian granular gland secretions for novel insulinotropic peptides. This study involved isolation and characterisation of biologically active peptides from the skin secretions of Rana palustris frogs. Crude secretions obtained by mild electrical stimulation from the dorsal skin surface were purified by reversed-phase HPLC on a semipreparative Vydac C18 column, yielding 80 fractions. These fractions were assayed for insulin-releasing activity using glucose-responsive BRIN-BD11 cells. Acute 20 min incubations were performed in Krebs Ringer bicarbonate buffer supplemented with 5.6 mmol/l glucose in the absence (control) and presence of various fractions. Fractions 29-54 and fractions 68-75 showed significant 2.0-6.5-fold increases in insulin-releasing activity (P<0.001). The fractions showing most prominent insulinotropic activity were further purified to single homogeneous peaks, which, on testing, evoked 1.5 2.8-fold increases in insulin release (P<0.001). The structures of the purified peptides were determined by mass spectrometry and N-terminal amino acid sequencing. Electrospray ionisation ion-trap mass spectrometry analysis revealed molecular masses of 2873.5-8560.4 Da. Sufficient material was isolated to determine the primary amino acid sequence of the 2873.5 Da peptide, revealing a 27 amino acid sequence, ALSILRGLEKLAKMGIALTNCKATKKC, repressing palustrin-1c. The database search for this peptide showed a 48% homology with brevinin-1, an antimicrobial peptide isolated from various Rana species, which itself stimulated insulin release from BRIN-BD11 cells in a concentration-dependent manner. In conclusion, the skin secretions of R. palustris frogs contain a novel class of peptides with insulin-releasing activity that merit further investigation. PMID- 15128284 TI - Modulation of steroidogenic enzymes by orphan nuclear transcriptional regulation may control diverse production of cortisol and androgens in the human adrenal. AB - The capacity of the adrenal to produce cortisol is controlled in part by 21 hydroxylase (CYP21) and the production of androgens by 17-hydroxylase/17-20-lyase (CYP17), in response to secretagogues including ACTH, angiotensin-II (A-II) and insulin. In this study we examined the capacity of human adrenocortical cells to produce cortisol and androgens in response to these secretagogues and their ability to regulate the expression of CYP21 and CYP17. In H-295 cells, forskolin and A-II were found to stimulate production of cortisol relative to androstenedione and a similar pattern of steroid production was noted in primary human adrenocortical cells. Both mRNA and protein expression of CYP21 was upregulated with forskolin and A-II alone and in combination, as detected by Northern and Western blotting. Whereas expression of CYP17 mRNA and protein was up regulated in the presence of forskolin and forskolin in combination with insulin. The ability of steroidogenic factor-1 (SF-1) and nur77 to regulate transcription of these enzymes was examined. Forskolin, A-II and insulin increased the protein expression of SF-1. Increased binding of SF-1 to its response element in the presence of forskolin, A-II and insulin was observed. Nur77 was expressed primarily in the zona glomerulosa and fasciculata. Increased protein expression of nur77 and the greatest binding of nur77 to its response element was seen when cells were stimulated with A-II in combination with forskolin. These data indicate that nur77 may preferentially regulate steroid enzyme genes relevant to cortisol production and thereby regulate differential cortisol and adrenal androgen production. PMID- 15128285 TI - Oral glutamine attenuates indomethacin-induced small intestinal damage. AB - The use of NSAIDs (non-steroidal anti-inflammatory drugs), although of great therapeutic value clinically, is limited by their tendency to cause mucosal damage in the gastrointestinal tract. In the small intestine, the effects these drugs have been shown to produce include inhibition of cyclo-oxygenase, mitochondrial dysfunction and free radical-induced oxidative changes, all of which contribute to the mucosal damage seen. Glutamine is a fuel preferentially used by enterocytes and is known to contribute to maintaining the integrity of these cells. In the present study, we investigated the effect of glutamine on indomethacin-induced changes in the small intestinal mucosa. Rats were given 2% glutamine or glutamic acid or isonitrogenous amino acids, glycine or alanine, in the diet for 7 days. Indomethacin was then administered orally at a dose of 40 mg/kg of body weight. After 1 h, the small intestine was removed and used for the measurement of parameters of oxidative stress and mitochondrial and BBM (brush border membrane) function. Evidence of oxidative stress was found in the mucosa of the small intestine of drug-treated rats, as indicated by significantly increased activity of xanthine oxidase (P < 0.001) and myeloperoxidase (P < 0.001), with corresponding decreases in the levels of several free radical scavenging enzymes and alpha-tocopherol (P < 0.001 in all cases). Levels of products of peroxidation were also significantly elevated (P < 0.001 for all the parameters measured). In addition, oxidative stress was evident in isolated intestinal mitochondria and BBMs (P < 0.001 for all the parameters measured), with associated alterations in function of these organelles (P < 0.001 for all the parameters measured). Supplementation of the diet with glutamine or glutamic acid prior to treatment with indomethacin produced significant amelioration in all the effects produced by the drug in the small intestine (P < 0.001 for all the parameters measured). Glycine and alanine were found to be much less effective in these respects. PMID- 15128287 TI - TIRF imaging of docking and fusion of single insulin granule motion in primary rat pancreatic beta-cells: different behaviour of granule motion between normal and Goto-Kakizaki diabetic rat beta-cells. AB - We imaged and analysed the motion of single insulin secretory granules near the plasma membrane in live pancreatic beta-cells, from normal and diabetic Goto Kakizaki (GK) rats, using total internal reflection fluorescence microscopy (TIRFM). In normal rat primary beta-cells, the granules that were fusing during the first phase originate from previously docked granules, and those during the second phase originate from 'newcomers'. In diabetic GK rat beta-cells, the number of fusion events from previously docked granules were markedly reduced, and, in contrast, the fusion from newcomers was still preserved. The dynamic change in the number of docked insulin granules showed that, in GK rat beta cells, the total number of docked insulin granules was markedly decreased to 35% of the initial number after glucose stimulation. Immunohistochemistry with anti insulin antibody observed by TIRFM showed that GK rat beta-cells had a marked decline of endogenous insulin granules docked to the plasma membrane. Thus our results indicate that the decreased number of docked insulin granules accounts for the impaired insulin release during the first phase of insulin release in diabetic GK rat beta-cells. PMID- 15128286 TI - Silica transport in the demosponge Suberites domuncula: fluorescence emission analysis using the PDMPO probe and cloning of a potential transporter. AB - Silicon is, besides oxygen, the most abundant element on earth. Only two taxa use this element as a major constituent of their skeleton, namely sponges (phylum Porifera) and unicellular diatoms. Results from combined cytobiological and molecularbiological techniques suggest that, in the demosponge Suberites domuncula, silicic acid is taken up by a transporter. Incubation of cells with the fluorescent silica tracer PDMPO [2-(4-pyridyl)-5-[[4-(2 dimethylaminoethylaminocarbamoyl)methoxy]phenyl]-oxazole] showed a response to silicic acid by an increase in fluorescence; this process is temperature dependent and can be blocked by DIDS (4,4-di-isothiocyanatostilbene-2,2 disulphonic acid). The putative NBC (Na+/HCO3-) transporter was identified, cloned and analysed. The deduced protein comprises all signatures characteristic of those molecules, and phylogenetic analysis also classifies it to the NBC transporter family. This cDNA was used to demonstrate that the expression of the gene is strongly up-regulated after treatment of cells with silicic acid. In situ hybridization demonstrated that the expression of the sponge transporter occurs in those cells that are located adjacent to the spicules (the skeletal element of the animal) or in areas in which spicule formation occurs. We conclude that this transporter is involved in silica uptake and have therefore termed it the NBCSA [Na+/HCO3-[Si(OH)4]] co-transporter. PMID- 15128288 TI - Topology of the plastid Ndh complex and its NDH-F subunit in thylakoid membranes. AB - We have investigated the topologies of Ndh (a plastid complex with NADH dehydrogenase activity) and its NDH-F subunit in thylakoids by trypsin and proteinase V8 digestion of both intact and Triton X-100-permeabilized barley thylakoids and identification of the products with antibodies against specific sequences of the NDH-A, NDH-K and NDH-F subunits. Antibody binding and protection against proteinases were also assayed. The analysis of the digestion products of NDH-F by immunodetection and matrix-assisted laser-desorption ionization-time-of flight allowed us to propose its membrane topology and to compare it with bioinformatic predictions and with that of the homologous subunit (ND5/NuoL/NQO12) of the respiratory complex I. Results indicate that the thylakoid Ndh complex may have an L-shaped structure, similar to that of respiratory complex I, with the hydrophilic arm orientated towards the stroma and the hydrophobic arm inserted into the thylakoid. NDH-A and NDH-K may be located at the bridge between the two arms. Similar to ND5/NuoL/NQO12 of complex I, NDH-F must be distally located in the hydrophobic arm. NDH-F would include up to 15 transmembrane helices and 14 hydrophilic regions. A conserved His-349 in the X transmembrane helix could be involved in H+ pumping. The conserved Thr-181 NDH-F, whose probable phosphorylation increases the activity of the Ndh complex, is located within the hydrophilic region between the V and VI transmembrane helices. PMID- 15128291 TI - Biochemical pharmacology of the vanilloid receptor TRPV1. An update. AB - There is mounting evidence that the vanilloid (capsaicin) receptor; transient receptor potential channel, vanilloid subfamily member 1 (TRPV1), is subjected to multiple interacting levels of control. The first level is by reversible phosphorylation catalyzed by intrinsic kinases (e.g. protein kinase A and C) and phosphatases (e.g. calcineurin), which plays a pivotal role in receptor sensitization vs. tachyphylaxis. In addition, this mechanism links TRPV1 to intracellular signaling by various important endogenous as well as exogenous substances such as bradykinin, ethanol, nicotin and insulin. It is not clear, however, whether phosphorylation per se is sufficient to liberate TRPV1 under the inhibitory control of phosphatydylinositol-4,5-bisphosphate. The second level of control is by forming TRPV1 heteromers and their association with putative regulatory proteins. The next level of regulation is by subcellular compartmentalization. The membrane form of TRPV1 functions as a nonselective cation channel. On the endoplasmic reticulum, TRPV1 is present in two differentially regulated forms, one of which is inositol triphosphate-dependent whereas the other is not. These three TRPV1 compartments provide a versatile regulation of intracellular Ca(2+) levels. Last, there is a complex and poorly understood regulation of TRPV1 activity via control of gene expression. Factors that downregulate TRPV1 expression include vanilloid treatment and growth factor (notably, nerve growth factor) deprivation. By contrast, TRPV1 appears to be upregulated during inflammatory conditions. Interestingly, following experimental nerve injury and in animal models of diabetic neuropathy TRPV1 is present on neurons that do not normally express TRPV1. Combined, these findings imply an important role for aberrant TRPV1 expression in the development of neuropathic pain and hyperalgesia. In humans, disease-related changes in TRPV1 expression have already been described (e.g. inflammatory bowel disease and irritable bowel syndrome). The mechanisms that regulate TRPV1 gene expression under pathological conditions are unknown but a better understanding of these pathways has obvious implications for rational drug development. PMID- 15128289 TI - Cellular effects of deoxynojirimycin analogues: inhibition of N-linked oligosaccharide processing and generation of free glucosylated oligosaccharides. AB - In the accompanying paper [Mellor, Neville, Harvey, Platt, Dwek and Butters (2004) Biochem. J. 381, 861-866] we treated HL60 cells with N-alk(en)yl deoxynojirimycin (DNJ) compounds to inhibit glucosphingolipid (GSL) biosynthesis and identified a number of non-GSL-derived, small, free oligosaccharides (FOS) most likely produced due to inhibition of the oligosaccharide-processing enzymes a-glucosidases I and II. When HL60 cells were treated with concentrations of N alk(en)ylated DNJ analogues that inhibited GSL biosynthesis completely, N-butyl- and N-nonyl-DNJ inhibited endoplasmic reticulum (ER) glucosidases I and II, but octadecyl-DNJ did not, probably due to the lack of ER lumen access for this novel, long-chain derivative. Glucosidase inhibition resulted in the appearance of free Glc1-3Man structures, which is evidence of Golgi glycoprotein endomannosidase processing of oligosaccharides with retained glucose residues. Additional large FOS was also detected in cells following a 16 h treatment with N butyl- and N-nonyl-DNJ. When these FOS structures (>30, including >20 species not present in control cells) were characterized by enzyme digests and MALDI-TOF (matrix-assisted laser-desorption ionization-time-of-flight) MS, all were found to be polymannose-type oligosaccharides, of which the majority were glucosylated and had only one reducing terminal GlcNAc (N-acetylglucosamine) residue (FOS GlcNAc1), demonstrating a cytosolic location. These results support the proposal that the increase in glucosylated FOS results from enzyme-mediated cytosolic cleavage of oligosaccharides from glycoproteins exported from the ER because of misfolding or excessive retention. Importantly, the present study characterizes the cellular properties of DNJs further and demonstrates that side-chain modifications allow selective inhibition of protein and lipid glycosylation pathways. This represents the most detailed characterization of the FOS structures arising from ER a-glucosidase inhibition to date. PMID- 15128292 TI - Molecular architecture of the vanilloid receptor. Insights for drug design. AB - The transient receptor potential channel vanilloid receptor subunit 1 (TRPV1) is a molecular integrator of physical and chemical stimuli in the peripheral nociceptor terminals. TRPV1 is an ionotropic channel that plays a critical role in both thermal nociception and inflammatory hyperalgesia. Structure-function relationships are providing fundamental insights of the modular architecture of this neuronal receptor. As a result, the molecular determinants that endow TRPV1 with its physiological properties, namely activation by heat, potentiation by extracellular acidic pH, and interaction with vanilloid-like compounds, as well as its permeation properties are being unveiled. This information can now be used to build up molecular models for the protein which, upon experimental validation, could be used as tools to thrust the target-oriented design of druggable TRPV1 ligands. PMID- 15128293 TI - Endovanilloids. Putative endogenous ligands of transient receptor potential vanilloid 1 channels. AB - Endovanilloids are defined as endogenous ligands of the transient receptor potential vanilloid type 1 (TRPV1) protein, a nonselective cation channel that belongs to the large family of TRP ion channels, and is activated by the pungent ingredient of hot chilli peppers, capsaicin. TRPV1 is expressed in some nociceptor efferent neurons, where it acts as a molecular sensor of noxious heat and low pH. However, the presence of these channels in various regions of the central nervous system, where they are not likely to be targeted by these noxious stimuli, suggests the existence of endovanilloids. Three different classes of endogenous lipids have been found recently that can activate TRPV1, i.e. unsaturated N-acyldopamines, lipoxygenase products of arachidonic acid and the endocannabinoid anandamide with some of its congeners. To classify a molecule as an endovanilloid, the compound should be formed or released in an activity dependent manner in sufficient amounts to evoke a TRPV1-mediated response by direct activation of the channel. To control TRPV1 signaling, endovanilloids should be inactivated within a short time-span. In this review, we will discuss, for each of the proposed endogenous ligands of TRPV1, their ability to act as endovanilloids in light of the criteria mentioned above. PMID- 15128294 TI - Prokaryotic and eukaryotic DNA helicases. Essential molecular motor proteins for cellular machinery. AB - DNA helicases are ubiquitous molecular motor proteins which harness the chemical free energy of ATP hydrolysis to catalyze the unwinding of energetically stable duplex DNA, and thus play important roles in nearly all aspects of nucleic acid metabolism, including replication, repair, recombination, and transcription. They break the hydrogen bonds between the duplex helix and move unidirectionally along the bound strand. All helicases are also translocases and DNA-dependent ATPases. Most contain conserved helicase motifs that act as an engine to power DNA unwinding. All DNA helicases share some common properties, including nucleic acid binding, NTP binding and hydrolysis, and unwinding of duplex DNA in the 3' to 5' or 5' to 3' direction. The minichromosome maintenance (Mcm) protein complex (Mcm4/6/7) provides a DNA-unwinding function at the origin of replication in all eukaryotes and may act as a licensing factor for DNA replication. The RecQ family of helicases is highly conserved from bacteria to humans and is required for the maintenance of genome integrity. They have also been implicated in a variety of human genetic disorders. Since the discovery of the first DNA helicase in Escherichia coli in 1976, and the first eukaryotic one in the lily in 1978, a large number of these enzymes have been isolated from both prokaryotic and eukaryotic systems, and the number is still growing. In this review we cover the historical background of DNA helicases, helicase assays, biochemical properties, prokaryotic and eukaryotic DNA helicases including Mcm proteins and the RecQ family of helicases. The properties of most of the known DNA helicases from prokaryotic and eukaryotic systems, including viruses and bacteriophages, are summarized in tables. PMID- 15128295 TI - Unraveling DNA helicases. Motif, structure, mechanism and function. AB - DNA helicases are molecular 'motor' enzymes that use the energy of NTP hydrolysis to separate transiently energetically stable duplex DNA into single strands. They are therefore essential in nearly all DNA metabolic transactions. They act as essential molecular tools for the cellular machinery. Since the discovery of the first DNA helicase in Escherichia coli in 1976, several have been isolated from both prokaryotic and eukaryotic systems. DNA helicases generally bind to ssDNA or ssDNA/dsDNA junctions and translocate mainly unidirectionally along the bound strand and disrupt the hydrogen bonds between the duplexes. Most helicases contain conserved motifs which act as an engine to drive DNA unwinding. Crystal structures have revealed an underlying common structural fold for their function. These structures suggest the role of the helicase motifs in catalytic function and offer clues as to how these proteins can translocate and unwind DNA. The genes containing helicase motifs may have evolved from a common ancestor. In this review we cover the conserved motifs, structural information, mechanism of DNA unwinding and translocation, and functional aspects of DNA helicases. PMID- 15128296 TI - The metabolic role and evolution of L-arabinitol 4-dehydrogenase of Hypocrea jecorina. AB - L-Arabinitol 4-dehydrogenase (Lad1) of the cellulolytic and hemicellulolytic fungus Hypocrea jecorina (anamorph: Trichoderma reesei) has been implicated in the catabolism of L-arabinose, and genetic evidence also shows that it is involved in the catabolism of D-xylose in xylitol dehydrogenase (xdh1) mutants and of D-galactose in galactokinase (gal1) mutants of H. jecorina. In order to identify the substrate specificity of Lad1, we have recombinantly produced the enzyme in Escherichia coli and purified it to physical homogeneity. The resulting enzyme preparation catalyzed the oxidation of pentitols (L-arabinitol) and hexitols (D-allitol, D-sorbitol, L-iditol, L-mannitol) to the same corresponding ketoses as mammalian sorbitol dehydrogenase (SDH), albeit with different catalytic efficacies, showing highest k(cat)/K(m) for L-arabinitol. However, it oxidized galactitol and D-talitol at C4 exclusively, yielding L-xylo-3-hexulose and D-arabino-3-hexulose, respectively. Phylogenetic analysis of Lad1 showed that it is a member of a terminal clade of putative fungal arabinitol dehydrogenase orthologues which separated during evolution of SDHs. Juxtapositioning of the Lad1 3D structure over that of SDH revealed major amino acid exchanges at topologies flanking the binding pocket for d-sorbitol. A lad1 gene disruptant was almost unable to grow on L-arabinose, grew extremely weakly on L-arabinitol, D talitol and galactitol, showed reduced growth on D-sorbitol and D-galactose and a slightly reduced growth on D-glucose. The weak growth on L-arabinitol was completely eliminated in a mutant in which the xdh1 gene had also been disrupted. These data show not only that Lad1 is indeed essential for the catabolism of L arabinose, but also that it constitutes an essential step in the catabolism of several hexoses; this emphasizes the importance of such reductive pathways of catabolism in fungi. PMID- 15128298 TI - Cloning, expression and characterization of two new IgE-binding proteins from the yeast Malassezia sympodialis with sequence similarities to heat shock proteins and manganese superoxide dismutase. AB - Malassezia sympodialis is an opportunistic yeast that colonizes human skin and may induce IgE and T cell reactivity in patients with atopic eczema/dermatitis syndrome (AEDS). Previously, we have cloned and expressed six recombinant allergens (rMala s 1 and rMala s 5 to rMala s 9) from this yeast. By combining high throughput screening and phage surface display techniques, 27 complete and partial IgE-binding clones of M. sympodialis have been identified. Here we enlarged the panel of recombinant M. sympodialis allergens by RACE-PCR, cloning and nucleotide sequencing to obtain the coding sequences of two new IgE-binding clones. The coding sequences of one of the clones showed similarity to the heat shock protein (HSP) family and the other to manganese superoxide dismutase (MnSOD), and both had a high degree of homology to human counterparts. The coding sequences were expressed in Escherichia coli as six-histidine tagged recombinant proteins and generated products with molecular masses of 86.1 kDa for HSP and 22.4 kDa for MnSOD. Their IgE-binding frequencies were shown to be 69% and 75%, respectively, to 28 sera from AEDS patients with serum IgE to M. sympodialis extract, indicating that HSP and MnSOD are major M. sympodialis allergens. In inhibition immunoblotting, M. sympodialis extract could inhibit the binding of serum IgE from AEDS patients to rHSP and rMnSOD in a concentration-dependent manner. The high frequency of sera from AEDS patients, showing IgE binding to both HSP and MnSOD, indicates that these allergens, designated Mala s 10 and Mala s 11, could play a role in AEDS. PMID- 15128297 TI - Actin-binding domain of mouse plectin. Crystal structure and binding to vimentin. AB - Plectin, a large and widely expressed cytolinker protein, is composed of several subdomains that harbor binding sites for a variety of different interaction partners. A canonical actin-binding domain (ABD) comprising two calponin homology domains (CH1 and CH2) is located in proximity to its amino terminus. However, the ABD of plectin is unique among actin-binding proteins as it is expressed in the form of distinct, plectin isoform-specific versions. We have determined the three dimensional structure of two distinct crystalline forms of one of its ABD versions (pleABD/2alpha) from mouse, to a resolution of 1.95 and 2.0 A. Comparison of pleABD/2alpha with the ABDs of fimbrin and utrophin revealed structural similarity between plectin and fimbrin, although the proteins share only low sequence identity. In fact, pleABD/2alpha has been found to have the same compact fold as the human plectin ABD and the fimbrin ABD, differing from the open conformation described for the ABDs of utrophin and dystrophin. Plectin harbors a specific binding site for intermediate filaments of various types within its carboxy-terminal R5 repeat domain. Our experiments revealed an additional vimentin-binding site of plectin, residing within the CH1 subdomain of its ABD. We show that vimentin binds to this site via the amino-terminal part of its rod domain. This additional amino-terminal intermediate filament protein binding site of plectin may have a function in intermediate filament dynamics and assembly, rather than in linking and stabilizing intermediate filament networks. PMID- 15128300 TI - Identification and expression of the first nonmammalian amyloid-beta precursor like protein APLP2 in the amphibian Xenopus laevis. AB - The Alzheimer's disease-linked amyloid-beta precursor protein (APP) belongs to a superfamily of proteins, which also comprises the amyloid-beta precursor-like proteins, APLP1 and APLP2. Whereas APP has been identified in both lower and higher vertebrates, thus far, APLP1 and 2 have been characterized only in human and rodents. Here we identify the first nonmammalian APLP2 protein in the South African claw-toed frog Xenopus laevis. The identity between the Xenopus and mammalian APLP2 proteins is approximately 75%, with the highest degree of conservation in a number of amino-terminal regions, the transmembrane domain and the cytoplasmic tail. Furthermore, amino acid residues known to be phosphorylated and glycosylated in mammalian APLP2 are conserved in Xenopus. The availability of the Xenopus APLP2 protein sequence allowed a phylogenetic analysis of APP superfamily members that suggested the occurrence of APP and preAPLP lineages with their separation predating the mammalian-amphibian split. As in mammals, Xenopus APLP2 mRNA was ubiquitously expressed and alternatively spliced forms were detected. However, the expression ratios between the mRNA forms in the various tissues examined were different between Xenopus and mammals, most prominently for the alternatively spliced forms containing the Kunitz protease inhibitor-coding region that were less abundantly expressed than the corresponding mammalian forms. Thus, the identification of APLP2 in Xenopus has revealed evolutionarily conserved regions that may help to delineate functionally important domains, and its overall high degree of conservation suggests an important role for this APP superfamily member. PMID- 15128299 TI - Ribosomal protein L22 inhibits regulation of cellular activities by the Epstein Barr virus small RNA EBER-1. AB - Epstein-Barr virus (EBV) is a potent mitogenic and antiapoptotic agent for B lymphocytes and is associated with several different types of human tumour. The abundantly expressed small viral RNA, EBER-1, binds to the growth inhibitory and pro-apoptotic protein kinase R (PKR) and blocks activation of the latter by double-stranded RNA. Recent evidence has suggested that expression of EBER-1 alone in EBV-negative B cells promotes a tumorigenic phenotype and that this may be related to inhibition of the pro-apoptotic effects of PKR. The ribosomal protein L22 binds to EBER-1 in virus-infected cells, but the significance of this has not previously been established. We report here that L22 and PKR compete for a common binding site on EBER-1. As a result of this competition, L22 interferes with the ability of the small RNA to inhibit the activation of PKR by dsRNA. Transient expression of EBER-1 in murine embryonic fibroblasts stimulates reporter gene expression and partially reverses the inhibitory effect of PKR. However, EBER-1 is also stimulatory when transfected into PKR knockout cells, suggesting an additional, PKR-independent, mode of action of the small RNA. Expression of L22 prevents both the PKR-dependent and -independent effects of EBER-1 in vivo. These results suggest that the association of L22 with EBER-1 in EBV-infected cells can attenuate the biological effects of the viral RNA. Such effects include both the inhibition of PKR and additional mechanism(s) by which EBER-1 stimulates gene expression. PMID- 15128301 TI - Identification and expression analysis of an IL-18 homologue and its alternatively spliced form in rainbow trout (Oncorhynchus mykiss). AB - A homologue of interleukin 18 has been identified from rainbow trout, Oncorhynchus mykiss. The trout IL-18 gene spans 3.7 kb and consists of six exons and five introns, sharing the same gene organization with its human counterpart. The putative translated protein is 199 amino acids in length with no predicted signal peptide. Analysis of the multiple sequence alignment reveals a conserved ICE cut site, resulting in a mature peptide of 162 amino acids. The trout IL-18 shares 41-45% similarity with known IL-18 molecules and contains an IL-1 family signature motif. It is constitutively expressed in a wide range of tissues including brain, gill, gut, heart, kidney, liver, muscle, skin and spleen. Transcription is not modulated by lipopolysaccharide, poly(I:C) or trout recombinant IL-1beta in primary head kidney leucocyte cultures and RTS-11 cells, a macrophage cell line. However, expression is downregulated by lipopolysaccharide and rIL-1beta in RTG-2 cells, a fibroblast-like cell line. An alternatively spliced form of IL-18 mRNA has also been found and translates into a 182 amino acid protein with a 17 amino acid deletion in the precursor region of the authentic form. This alternatively spliced form is also widely expressed although much lower than the authentic form. Interestingly, its expression is upregulated by lipopolysaccharide and poly(I:C), but is not affected by rIL-1beta in RTG-2 cells. The present study suggests that alternative splicing may play an important role in regulating IL-18 activities in rainbow trout. PMID- 15128302 TI - A (1-->3)-beta-D-glucan recognition protein from the sponge Suberites domuncula. Mediated activation of fibrinogen-like protein and epidermal growth factor gene expression. AB - Sponges (phylum Porifera) live in a symbiotic relationship with microorganisms, primarily bacteria. Until now, molecular proof for the capacity of sponges to recognize fungi in the surrounding aqueous milieu has not been available. Here we demonstrate, for the demosponge Suberites domuncula (Porifera, Demospongiae, Hadromerida), a cell surface receptor that recognizes (1-->3)-beta-D-glucans, e.g. curdlan or laminarin. This receptor, the (1-->3)-beta-D-glucan-binding protein, was identified and its cDNA analysed. The gene coding for the 45 kDa protein was found to be upregulated in tissue after incubation with carbohydrate. Simultaneously with the increased expression of this gene, two further genes showed an elevated steady state level of expression; one codes for a fibrinogen like protein and the other for the epidermal growth factor precursor. Expression of the (1-->3)-beta-D-glucan-binding protein and the fibrinogen-like protein occurred in cells on the sponge surface, in the pinacoderm. By Western blotting, the product of the fibrinogen-like protein gene was identified, the recombinant protein isolated, and antibodies raised to this protein. Their application revealed that a 5 kDa factor is produced, which is apparently processed from the 77 kDa epidermal growth factor precursor. Finally, we provided evidence that a tyrosine kinase pathway is initiated in response to exposure to D-glucan; its phosphorylation activity could be blocked by aeroplysinin. In turn, the increased expression of the downstream genes was suppressed. We conclude that sponges possess a molecular mechanism for recognizing fungi via the d-glucan carbohydrates on their surfaces. PMID- 15128303 TI - Structure and topology of the transmembrane domain 4 of the divalent metal transporter in membrane-mimetic environments. AB - The divalent metal transporter (DMT1) is a 12-transmembrane domain protein responsible for dietary iron uptake in the duodenum and iron acquisition from transferrin in peripheral tissues. The transmembrane domain 4 (TM4) of DMT1 has been shown to be crucial for its biological function. Here we report the 3D structure and topology of the DMT1-TM4 peptide by NMR spectroscopy with simulated annealing calculations in membrane-mimetic environments, e.g. 2,2,2 trifluoroethanol and SDS micelles. The 3D structures of the peptide are similar in both environments, with nonordered and flexible N- and C-termini flanking an ordered helical region. The final set of the 16 lowest energy structures is particularly well defined in the region of residues Leu9-Phe20 in 2,2,2 trifluoroethanol, with a mean pairwise root mean square deviation of 0.23 +/- 0.10 A for the backbone heavy atoms and 0.82 +/- 0.17 A for all heavy atoms. In SDS micelles, the length of the helix is dependent on pH values. In particular, the C-terminus becomes well-structured at low pH (4.0), whereas the N-terminal segment (Arg1-Gly7) is flexible and poorly defined at all pH values studied. The effects of 12-doxylPtdCho spin-label and paramagnetic metal ions on NMR signal intensities demonstrated that both the N-terminus and helical region of the TM4 are embedded into the interior of SDS micelles. Unexpectedly, we observed that amide protons exchanged much faster in SDS than in 2,2,2-trifluoroethanol, indicating that there is possible solvent accessibility in the structure. The paramagnetic metal ions broaden NMR signals from residues both situated in aqueous phase and in the helical region. From these results we speculate that DMT1-TM4s may self-assemble to form a channel through which metal ions are likely to be transported. These results might provide an insight into the structure function relationship for the integral DMT1. PMID- 15128304 TI - Purification and characterization of Helicobacter pylori arginase, RocF: unique features among the arginase superfamily. AB - The urea cycle enzyme arginase (EC 3.5.3.1) hydrolyzes l-arginine to l-ornithine and urea. Mammalian arginases require manganese, have a highly alkaline pH optimum and are resistant to reducing agents. The gastric human pathogen, Helicobacter pylori, also has a complete urea cycle and contains the rocF gene encoding arginase (RocF), which is involved in the pathogenesis of H. pylori infection. Its arginase is specifically involved in acid resistance and inhibits host nitric oxide production. The rocF gene was found to confer arginase activity to Escherichia coli; disruption of plasmid-borne rocF abolished arginase activity. A translationally fused His(6)-RocF was purified from E. coli under nondenaturing conditions and had catalytic activity. Remarkably, the purified enzyme had an acidic pH optimum of 6.1. Both purified arginase and arginase containing H. pylori extracts exhibited optimal catalytic activity with cobalt as a metal cofactor; manganese and nickel were significantly less efficient in catalyzing the hydrolysis of arginine. Viable H. pylori or E. coli containing rocF had significantly more arginase activity when grown with cobalt in the culture medium than when grown with manganese or no divalent metal. His(6)-RocF arginase activity was inhibited by low concentrations of reducing agents. Antibodies raised to purified His(6)-RocF reacted with both H. pylori and E. coli extracts containing arginase, but not with extracts from rocF mutants of H. pylori or E. coli lacking the rocF gene. The results indicate that H. pylori RocF is necessary and sufficient for arginase activity and has unparalleled features among the arginase superfamily, which may reflect the unique gastric ecological niche of this organism. PMID- 15128305 TI - Functional transitions of F0F1-ATPase mediated by the inhibitory peptide IF1 in yeast coupled submitochondrial particles. AB - The mechanism of inhibition of yeast F(0)F(1)-ATPase by its naturally occurring protein inhibitor (IF1) was investigated in submitochondrial particles by studying the IF1-mediated ATPase inhibition in the presence and absence of a protonmotive force. In the presence of protonmotive force, IF1 added during net NTP hydrolysis almost completely inhibited NTPase activity. At moderate IF1 concentration, subsequent uncoupler addition unexpectedly caused a burst of NTP hydrolysis. We propose that the protonmotive force induces the conversion of IF1 inhibited F(0)F(1)-ATPase into a new form having a lower affinity for IF1. This form remains inactive for ATP hydrolysis after IF1 release. Uncoupling simultaneously releases ATP hydrolysis and converts the latent form of IF1-free F(0)F(1)-ATPase back to the active form. The relationship between the different steps of the catalytic cycle, the mechanism of inhibition by IF1 and the interconversion process is discussed. PMID- 15128306 TI - Correlation between functional and structural changes of reduced and oxidized trout hemoglobins I and IV at different pHs. A circular dichroism study. AB - Circular dichroism (CD) spectra of two major hemoglobin components (Hb), HbI and HbIV, from Oncorhyncus mykiss (formerly Salmo irideus) trout were evaluated in the range 250-600 nm. HbI is characterized by a complete insensitivity to pH changes, while HbIV presents the Root effect. Both reduced [iron(II) or oxy] and oxidized (met) forms of the two proteins were studied at different pHs, 7.8 and 6.0, to obtain information about the pH effects on the structural features of these hemoglobins. Data obtained show that oxy and met-HbI are almost insensitive to pH decrease, remaining in the R conformational state also at low pH. On the contrary, the pH decrease induces similar structural changes, characteristics of ligand dissociation and R-->T transition, both in the reduced and in the oxidized HbIV. The structural changes, monitored by CD, are compared with the peroxidative activity of iron(II)-Hb and met-Hb forms and with the superoxide anion scavenger capacity of the proteins. PMID- 15128307 TI - Rate-determining step of butyrylcholinesterase-catalyzed hydrolysis of benzoylcholine and benzoylthiocholine. Volumetric study of wild-type and D70G mutant behavior. AB - The rate-limiting step for hydrolysis of the positively charged oxoester benzoylcholine (BzCh) by human butyrylcholinesterase (BuChE) is deacylation (k(3)), whereas it is acylation (k(2)) for hydrolysis of the homologous thioester benzoylthiocholine (BzSCh). Steady-state hydrolysis of BzCh and BzSCh by wild type BuChE and its peripheral anionic site mutant D70G was investigated at different hydrostatic pressures, which allowed determination of volume changes associated with substrate binding, and the activation volumes for the chemical steps. A differential nonlinear pressure-dependence of the catalytic parameters for hydrolysis of both substrates by both enzymes was shown. Nonlinearity of the plots may be explained in terms of compressibility changes or rate-limiting changes. To distinguish between these two possibilities, enzyme phosphorylation by diisopropylfluorophosphate (DFP) in the presence of substrate (BzSCh) under pressure was studied. There was no pressure dependence of volume changes for DFP binding or for phosphorylation of either wild-type or D70G. Analysis of the pressure dependence for steady-state hydrolysis of substrates, and for phosphorylation by DFP provided evidence that no enzyme compressibility changes occurred during the catalyzed reactions. Thus, the nonlinear pressure dependence of substrate hydrolysis reflects changes in the rate-limiting step with pressure. Change in rate-determining step occurred at a pressure of 100 MPa for hydrolysis of BzCh by wild-type and at 75 MPa for D70G. For hydrolysis of BzSCh the change occurred at higher pressures because k(2) << k(3) at atmospheric pressure for this substrate. Elementary volume change contributions upon initial binding, productive binding, acylation and deacylation were calculated from the pressure differentiation of kinetic constants. This analysis shed light on the molecular events taking place along the hydrolysis pathways of BzCh and BzSCh by wild-type BuChE and the D70G mutant. In addition, volume change differences between wild type and D70G provided new evidence that residue D70 in the peripheral site controls hydration of the active site gorge and the dynamics of the water molecule network during catalysis. Finally, a steady-state kinetic study of the oxyanion hole mutant (G117H) showed that substitution of the ethereal sulfur for oxygen in the substrate alters the final adjustment of substrate in the active site and stabilization of the acylation transition state. PMID- 15128308 TI - Anti- and pro-oxidant effects of quercetin in copper-induced low density lipoprotein oxidation. Quercetin as an effective antioxidant against pro-oxidant effects of urate. AB - We recently reported that, depending on its concentration, urate is either a pro- or an antioxidant in Cu(2+)-induced low-density lipoprotein (LDL) oxidation. We also previously demonstrated an antioxidant synergy between urate and some flavonoids in the Cu(2+)-induced oxidation of diluted serum. As a result, the effect of the flavonoid quercetin on the Cu(2+)-induced oxidation of isolated LDL has been studied either in the presence or absence of urate. We demonstrate that, like urate, quercetin alone, at low concentration, exhibits a pro-oxidant activity. The pro-oxidant behavior depends on the Cu(2+) concentration but it is not observed at high Cu(2+) concentration. When compared with urate, the switch between the pro- and the antioxidant activities occurs at much lower quercetin concentrations. As for urate, the pro-oxidant character of quercetin is related to its ability to reduce Cu(2+) with the formation of semioxidized quercetin and Cu(+) with an expected yield larger than that obtained with urate owing to a more favorable redox potential. It is also shown that the pro-oxidant activity of urate can be inhibited by quercetin. An electron transfer between quercetin and semioxidized urate leading to the repair of urate could account for this observation as suggested by recently published pulse radiolysis data. It is anticipated that the interactions between quercetin-Cu(2+)-LDL and urate, which are tightly controlled by their respective concentration, determine the balance between the pro- and antioxidant behaviors. Moreover, as already observed with other antioxidants, it is demonstrated that quercetin alone behaves as a pro oxidant towards preoxidized LDL. PMID- 15128309 TI - Characterization of a digestive carboxypeptidase from the insect pest corn earworm (Helicoverpa armigera) with novel specificity towards C-terminal glutamate residues. AB - Carboxypeptidases were purified from guts of larvae of corn earworm (Helicoverpa armigera), a lepidopteran crop pest, by affinity chromatography on immobilized potato carboxypeptidase inhibitor, and characterized by N-terminal sequencing. A larval gut cDNA library was screened using probes based on these protein sequences. cDNA HaCA42 encoded a carboxypeptidase with sequence similarity to enzymes of clan MC [Barrett, A. J., Rawlings, N. D. & Woessner, J. F. (1998) Handbook of Proteolytic Enzymes. Academic Press, London.], but with a novel predicted specificity towards C-terminal acidic residues. This carboxypeptidase was expressed as a recombinant proprotein in the yeast Pichia pastoris. The expressed protein could be activated by treatment with bovine trypsin; degradation of bound pro-region, rather than cleavage of pro-region from mature protein, was the rate-limiting step in activation. Activated HaCA42 carboxypeptidase hydrolysed a synthetic substrate for glutamate carboxypeptidases (FAEE, C-terminal Glu), but did not hydrolyse substrates for carboxypeptidase A or B (FAPP or FAAK, C-terminal Phe or Lys) or methotrexate, cleaved by clan MH glutamate carboxypeptidases. The enzyme was highly specific for C-terminal glutamate in peptide substrates, with slow hydrolysis of C-terminal aspartate also observed. Glutamate carboxypeptidase activity was present in larval gut extract from H. armigera. The HaCA42 protein is the first glutamate-specific metallocarboxypeptidase from clan MC to be identified and characterized. The genome of Drosophila melanogaster contains genes encoding enzymes with similar sequences and predicted specificity, and a cDNA encoding a similar enzyme has been isolated from gut tissue in tsetse fly. We suggest that digestive carboxypeptidases with sequence similarity to the classical mammalian enzymes, but with specificity towards C-terminal glutamate, are widely distributed in insects. PMID- 15128310 TI - Transport of the phosphonodipeptide alafosfalin by the H+/peptide cotransporters PEPT1 and PEPT2 in intestinal and renal epithelial cells. AB - The interaction of the antibacterial phosphonodipeptide alafosfalin with mammalian H(+)/peptide cotransporters was studied in Caco-2 cells, expressing the low-affinity intestinal type peptide transporter 1 (PEPT1), and SKPT cells, expressing the high-affinity renal type peptide transporter 2 (PEPT2). Alafosfalin strongly inhibited the uptake of [(14)C]glycylsarcosine with K(i) values of 0.19 +/- 0.01 mm and 0.07 +/- 0.01 mm for PEPT1 and PEPT2, respectively. Saturation kinetic studies revealed that in both cell types alafosfalin affected only the affinity constant (K(t)) but not the maximal velocity (V(max)) of glycylsarcosine (Gly-Sar) uptake. The inhibition constants and the competitive nature of inhibition were confirmed in Dixon-type experiments. Caco-2 cells and SKPT cells were also cultured on permeable filters: apical uptake and transepithelial apical to basolateral flux of [(14)C]Gly-Sar across Caco-2 cell monolayers were reduced by alafosfalin (3 mm) by 73%. In SKPT cells, uptake of [(14)C]Gly-Sar but not flux was inhibited by 61%. We found no evidence for an inhibition of the basolateral to apical uptake or flux of [(14)C]Gly-Sar by alafosfalin. Alafosfalin (3 mm) did not affect the apical to basolateral [(14)C]mannitol flux. Determined in an Ussing-type experiment with Caco-2 cells cultured in Snapwells trade mark, alafosfalin increased the short circuit current through Caco-2 cell monolayers. We conclude that alafosfalin interacts with both H(+)/peptide symporters and that alafosfalin is actively transported across the intestinal epithelium in a H(+)-symport, explaining its oral availability. The results also demonstrate that dipeptides where the C terminal carboxyl group is substituted by a phosphonic function represent high affinity substrates for mammalian H(+)/peptide cotransporters. PMID- 15128311 TI - The first representative of glycosylated three-fingered toxins. Cytotoxin from the Naja kaouthia cobra venom. AB - There are different glycosylated proteins in snake venoms, but no glycosylated representatives of a large family of three-fingered toxins have previously been detected. A new glycoprotein was isolated from the venom of the Thai cobra Naja kaouthia. MALDI MS of the glycoprotein contained an array of peaks in the range from approximately 8900 to approximately 9400 Da indicating its microheterogeneity. Carbohydrate analysis showed the presence of mannose, galactose, N-acetylglucosamine, fucose and neuraminic acid. The N-terminal sequence of the glycoprotein was identical to that of cytotoxin 3 (CX3) from N. kaouthia, and CD spectra of the glycoprotein and CX3 were almost the same. Cleavage of a glycan moiety by N-glycosidase F gave a protein of molecular mass practically coinciding with that of CX3. MALDI MS of the tryptic digest of reduced glycoprotein S-pyridylethylated at cysteine residues, contained peaks corresponding to all tryptic fragments of CX3, with the exception of fragment 24 30. The peak corresponding to this peptide appeared in the mass-spectrum of similarly treated deglycosylated glycoprotein. These data show that the potential N-glycosylation site at Asn29 in CX3 is utilized for glycan attachment and that the glycoprotein is glycosylated CX3. In vivo toxicity of the glycoprotein to the cricket Gryllus assimilis was twofold lower than that of CX3. The cytotoxic activity of the glycoprotein towards HL60 cells was about two orders of magnitude lower than that of CX3, but could be made equal to the CX3 cytotoxicity by deglycosylation. Thus for the first time we have isolated a glycosylated three fingered snake venom toxin wherein glycosylation appears to modulate its biological activity. PMID- 15128312 TI - Lipids and glycosphingolipids in caveolae and surrounding plasma membrane of primary rat adipocytes. AB - We have made a comprehensive and quantitative analysis of the lipid composition of caveolae from primary rat fat cells and compared the composition of plasma membrane inside and outside caveolae. We isolated caveolae from purified plasma membranes using ultrasonication in carbonate buffer to disrupt the membrane, or extraction with nonionic detergent, followed by density gradient ultracentrifugation. The carbonate-isolated caveolae fraction was further immunopurified using caveolin antibodies. Carbonate-isolated caveolae were enriched in cholesterol and sphingomyelin, and the concentration was three- and twofold higher, respectively, in caveolae compared to the surrounding plasma membrane. The concentration of glycerophospholipids was similar suggesting that glycerophospholipids constitute a constant core throughout the plasma membrane. The composition of detergent-insoluble fractions of the plasma membrane was very variable between preparations, but strongly enriched in sphingomyelin and depleted of glycerophospholipids compared to carbonate-isolated caveolae; indicating that detergent extraction is not a suitable technique for caveolae preparation. An average adipocyte caveola contained about 22 x 10(3) molecules of cholesterol, 7.5 x 10(3) of sphingomyelin and 23 x 10(3) of glycerophospholipid. The glycosphingolipid GD3 was highly enriched in caveolae, whereas GM3, GM1 and GD1a were present inside as well as outside the caveolae membrane. GD1b, GT1b, GM2, GQ1b, sulfatide and lactosylceramide sulfate were not detected in caveolae. PMID- 15128313 TI - Biophysical characterization of the interaction of Limulus polyphemus endotoxin neutralizing protein with lipopolysaccharide. AB - Endotoxin-neutralizing protein (ENP) of the horseshoe crab is one of the most potent neutralizers of endotoxins [bacterial lipopolysaccharide (LPS)]. Here, we report on the interaction of LPS with recombinant ENP using a variety of physical and biological techniques. In biological assays (Limulus amebocyte lysate and tumour necrosis factor-alpha induction in human mononuclear cells), ENP causes a strong reduction of the immunostimulatory ability of LPS in a dose-dependent manner. Concomitantly, the accessible negative surface charges of LPS and lipid A (zeta potential) are neutralized and even converted into positive values. The gel to liquid crystalline phase transitions of LPS and lipid A shift to higher temperatures indicative of a rigidification of the acyl chains, however, the only slight enhancement of the transition enthalpy indicates that the hydrophobic moiety is not strongly disturbed. The aggregate structure of lipid A is converted from a cubic into a multilamellar phase upon ENP binding, whereas the secondary structure of ENP does not change due to the interaction with LPS. ENP contains a hydrophobic binding site to which the dye 1-anilino-8-sulfonic acid binds at a K(d) of 19 micro m, which is displaced by LPS. Because lipopolysaccharide-binding protein (LBP) is not able to bind to LPS when ENP and LPS are preincubated, tight binding of ENP to LPS can be deduced with a K(d) in the low nonomolar range. Importantly, ENP is able to incorporate by itself into target phospholipid liposomes, and is also able to mediate the intercalation of LPS into the liposomes thus acting as a transport protein in a manner similar to LBP. Thus, LPS-ENP complexes might enter target membranes of immunocompetent cells, but are not able to activate due to the ability of ENP to change LPS aggregates from an active into an inactive form. PMID- 15128314 TI - The sex of the individual as a factor in allergic contact dermatitis. AB - Exogenous and endogenous factors have been implicated in allergic contact dermatitis (ACD); this review explores the sex of the individual as a possible endogenous factor. While there is a clinical impression that women are more skin reactive than men, upon review this hypothesis appears oversimplified. This review explores sex differences in controlled testing of adult ACD, occupational ACD and juvenile ACD. Further, geographical location and socioeconomic and cultural factors in relation to sex differences in allergic contact dermatitis are discussed. We conclude that female sex could be a factor predisposing to allergic contact dermatitis not so much because of possible differences in intrinsic skin characteristics between the sexes, but more because of different exposure patterns. This conclusion has practical consequences regarding product labelling requirements, occupational risk assessment and legislation. PMID- 15128315 TI - Occupational immunologic contact urticaria from pine processionary caterpillar (Thaumetopoea pityocampa): experience in 30 cases. AB - Cutaneous lesions caused by the pine processionary caterpillar Thaumetopoea pityocampa (TP) are frequent in pinewood areas. In the present study, 30 patients diagnosed with occupational immunologic urticaria from this caterpillar were included. Immediate hypersensitivity was demonstrated by performing prick and IgE immunoblotting tests. Workers were grouped according to their common tasks. Occupations at risk of exposure to TP were pine-cone collectors/woodcutters (14), farmers/stockbreeders (8), other forestry personnel (4), construction workers (2), residential gardeners (1) and entomologists (1). Besides contact urticaria, angioedema (60%), papular lesions of several days of evolution (30%) and anaphylactic reactions (40%) were also detected. The most frequently detected molecular weight bands by immunoblot were 15 (70%), 17 (57%) and 13 kDa (50%). The appearance of isolated bands corresponds with the least serious cases. Only 8 subjects had bands higher than 33 kDa, which was present in the 3 most severe cases of anaphylactic reactions. By presenting these cases, we wish to offer the largest series reported so far of occupational immunologic contact urticaria caused by TP. We include the first cases described in certain occupations, some of them not directly related to forestry work. Pine-cone or resin collectors, woodcutters, farmers and stockbreeders were the most frequently and severely affected workers. PMID- 15128316 TI - Contact allergy to fragrances: frequencies of sensitization from 1996 to 2002. Results of the IVDK*. AB - Increasing frequencies of sensitization to the fragrance mix (FM) have been acknowledged as a serious problem for many years. It is well known that the single compounds (SCs) of the FM contribute differently to the FM patch rest reactions. In this study, we were interested in the time trends of the FM, the SCs, Myroxylon pereirae resin (MP; balsam of Peru) and oil of turpentine (OT) as possible further indicators of perfume allergy and analysed the data collected by the Information Network of Departments of Dermatology multicentre project from 1996 to 2002. During the study period (1996-2002), the FM [8% petrolatum (pet.)], MP (25% pet.) and OT (1% pet.) were tested in 59,298, 59,334 and 59,478 patients, respectively. SCs were tested in a selected group of patients, ranging from n = 1083 to n = 1924 per year. A significant increase in the proportions of patients with positive reactions to FM, MP and OT between 1996 and 1998 is noted, and a significant decline from 1999 to 2002 (Cochrane Armitage trend test, P < 0.0001). The highest frequency of sensitization to the FM was 13.1% in 1999, and the lowest 7.8% in 2002. The number of concomitant reactions to OT, a surrogate marker for terpenes, in FM-positive patients was significantly increased between 1997 and 1999. Reactions to SCs in FM-positive patients were observed in 29.9% (oak moss absolute) to 5.9% (geraniol). There was no time trend in reactions to SCs, although the relative share was increased for isoeugenol, cinnamic aldehyde and geraniol in 1999. In summary, we report for the first time, a significant decline in sensitization to the FM, very probably due to a reduced exposure (less potent allergens used in fine fragrances, possibly less use of natural ingredient based cosmetics and lowered use concentration of important fragrance allergens). The differences in ranking of SCs could stimulate (a) a redefinition of the FM and (b) a differentiated preventive and regulatory approach, with oak moss and isoeugenol being regulated strictly by prohibition, concentration limits further reconsidered and/or health warnings and clearly less noxious substances like geraniol treated less restrictively. PMID- 15128317 TI - Occupational skin diseases among dental nurses. AB - Out of 923 female dental nurses in the Helsinki district, 799 were interviewed using a computer-assisted telephone interview. A structured questionnaire was used to inquire about skin, respiratory symptoms, atopy, work history and methods, and exposure at work. The 328 nurses, who reported work-related dermatitis on their hands, forearms or face, were invited to an interview by an occupational physician; 245 nurses participated. 31 nurses had previously been diagnosed with an occupational skin disease (OSD). 133 nurses with a suspected OSD were selected for further clinical examinations with prick and patch testing. Among the 107 nurses examined, 22 new cases of OSD were diagnosed. There were altogether 29 cases of allergic contact dermatitis, 15 of contact urticaria, 12 of irritant contact dermatitis, and 1 case of onychomycosis. Rubber chemicals and natural rubber latex (NRL) in protective gloves, as well as dental-restorative plastic materials [(meth)acrylates], were the most common causes of allergy. 42% of the OSD cases in the studied population had been missing from the statistics (Finnish Register of Occupational Diseases). Plastic gloves or NRL gloves with a low-protein content are recommended for dental work. Skin exposure to (meth)acrylates should be avoided. PMID- 15128318 TI - Changing patterns of contact allergy in chronic inflammatory ear disease. AB - Chronic inflammatory ear disease comprises a wide range of conditions. Although contact allergy to topical medicaments is common in such conditions, the causes of these reactions have not been examined in a large series for over a decade. Our aim was to investigate the major sensitizers recorded in 179 patients over the last 17 years. Medicament contact allergic dermatitis was diagnosed in 45 patients (25%), of whom 34 (76%) demonstrated a relevant allergy to neomycin, 28 (62%) to framycetin, 11 (31%) to gentamicin, 8 (18%) to quinoline mix and 5 (11%) to caine mix. Of all neomycin-allergic patients, 28 (76%) were also allergic to framycetin and 12 (43%) to gentamicin. Comparing the first 60 patients (1985-94) to the most recent 60 (1998-2002) identified a significant recent reduction in the number of patients allergic to neomycin (P = 0.002) and framycetin (P = 0.050), with a non-significant reduction in the total number diagnosed as medicament-allergic (P = 0.066). We have therefore demonstrated a high frequency of medicament contact allergy in chronic inflammatory ear disease, supporting the routine use of patch testing in these patients. We have also shown a significant recent reduction in the number of such patients with allergy to neomycin and framycetin. PMID- 15128319 TI - Eyelid dermatitis: a report of 215 patients. AB - Between April 2001 and October 2003, 215 persons presented with eyelid dermatitis for the first time. They ranged in age from 4 months to 95 years, with an average age of 49.6 years and a median age of 51 years. There were 173 females and 42 males. 165 of 215 had allergic contact dermatitis (ACD) and another 9 had protein contact dermatitis without relevant positive patch tests. Atopic eczema comprised 37 of 215 (17%), but 33 of 37 also had contact allergies. Seborrheic dermatitis, psoriasis or both were found in 35 (16%). Sources of ACD included personal care products in 54 (25%), including 12 with at least 1 positive patch test to cosmetic applicators and 12 sensitive to at least 1 botanical ingredient. Allergy to artificial nails and/or nail lacquer occurred in 18 of 215 (8%). 5 persons had rosacea or periorbital dermatitis, and 2 had dermatomyositis. Other causes included bacterial, fungal and viral infections, some of which were quite unusual. The evaluation of persons presenting with eyelid dermatitis remains a relatively complex but rewarding discipline. PMID- 15128321 TI - Widespread vitiligo after erythroderma caused by photosensitivity to flutamide. PMID- 15128320 TI - Further evaluation of quantitative structure--activity relationship models for the prediction of the skin sensitization potency of selected fragrance allergens. AB - Fragrance substances represent a very diverse group of chemicals; a proportion of them are associated with the ability to cause allergic reactions in the skin. Efforts to find substitute materials are hindered by the need to undertake animal testing for determining both skin sensitization hazard and potency. One strategy to avoid such testing is through an understanding of the relationships between chemical structure and skin sensitization, so-called structure-activity relationships. In recent work, we evaluated 2 groups of fragrance chemicals -- saturated aldehydes and alpha,beta-unsaturated aldehydes. Simple quantitative structure-activity relationship (QSAR) models relating the EC3 values [derived from the local lymph node assay (LLNA)] to physicochemical properties were developed for both sets of aldehydes. In the current study, we evaluated an additional group of carbonyl-containing compounds to test the predictive power of the developed QSARs and to extend their scope. The QSAR models were used to predict EC3 values of 10 newly selected compounds. Local lymph node assay data generated for these compounds demonstrated that the original QSARs were fairly accurate, but still required improvement. Development of these QSAR models has provided us with a better understanding of the potential mechanisms of action for aldehydes, and hence how to avoid or limit allergy. Knowledge generated from this work is being incorporated into new/improved rules for sensitization in the expert toxicity prediction system, deductive estimation of risk from existing knowledge (DEREK). PMID- 15128322 TI - Systemic contact dermatitis from nickel associated with orthodontic appliances. PMID- 15128323 TI - Contact dermatitis due to eggplant. PMID- 15128324 TI - Occupational allergic contact dermatitis from olive oil in pizza making. PMID- 15128325 TI - Schirmer I testing in eyelid dermatitis. PMID- 15128326 TI - Occupational allergic contact dermatitis caused by 1,2-benzisothiazolin-3-one in a varnish maker, followed by sensitization to benzalkonium chloride in Oilatum Plus bath additive. PMID- 15128327 TI - Erythema ab igne due to a laptop computer. PMID- 15128328 TI - Concurrent rubber allergy among workers in a silicone-wafer-manufacturing plant. PMID- 15128329 TI - Aluminium and milk allergies in an adult. PMID- 15128335 TI - Medicare prescription drugs and reform: what the new law will mean to gastroenterologists and their patients. PMID- 15128336 TI - Predictors of outcome in an open label, therapeutic trial of high-dose omeprazole in laryngitis. AB - BACKGROUND: Gastroesophageal reflux is implicated in some cases of laryngitis. There are no established predictors of response to acid suppression therapy in suspected reflux laryngitis. AIM: In a population with laryngitis, the aim is to determine whether (a) omeprazole 20 mg tds (3 months) improves symptoms and laryngitis, and (b) the outcome in response to potent acid suppression can be predicted by esophageal and/or pharyngeal parameters during ambulatory pH monitoring or by other pretreatment variables. METHODS: From the 70 consecutive patients with laryngitis screened, 20 patients met the inclusion criteria (dysphonia >3 months; laryngoscopically demonstrated laryngitis); and 50 patients were excluded because of one or more criteria indicating alternative causes for laryngeal injury. The primary outcome measure was improvement of at least one level in a 4-point laryngitis grading at 3 months. Twenty-four-hour dual, pharyngo-esophageal pH monitoring was performed at baseline. Secondary outcomes (symptom questionnaire; computerized voice analysis) were measured at baseline, and at 6 and 12 wk. RESULTS: Response rates at 6 and 12 wk were 47% and 63%, respectively. GERD symptoms (heartburn (p= 0.03) and regurgitation (p= 0.0001)) improved. However, neither baseline GERD symptoms nor endoscopic findings predicted laryngoscopic or symptomatic response. Neither baseline laryngitis grade (p= 0.46) nor esophageal acid exposure on pH testing (p= 0.3) predicted outcome. Four of 20 patients demonstrated pharyngeal regurgitation on pH testing, all four of whom responded to potent acid suppression (p= 0.2). Computerized voice measures were not predictive of outcome, although fundamental frequency (Fo) was inversely related to baseline laryngoscopic grade. CONCLUSION: In a carefully defined population of patients with laryngitis (a) 63% have a laryngoscopic response to 3 months of potent acid suppression without significant improvement in laryngeal symptoms; (b) neither voice measures, esophageal acid exposure time, symptoms nor severity of laryngitis predict outcome; and (c) although numbers were small, all patients with a positive pharyngeal pH study responded to therapy and pharyngeal pH-metry may prove useful; (4) available evidence supports an empiric trial of high-dose proton pump inhibitors (PPI), for at least 12 wk, as the initial diagnostic step for suspected reflux laryngitis. PMID- 15128337 TI - Laryngitis and gastroesophageal reflux disease: increasing prevalence or poor diagnostic tests? AB - Laryngeal signs and symptoms are often associated with gastroesophageal reflux disease (GERD). However, such diagnoses presume that laryngeal findings may be specific for GERD. However, neither laryngoscopy, EGD or pH monitoring are specific tests for identifying GERD related laryngitis. Non-placebo controlled trials often show clinical benefit from proton pump inhibitor therapy; however, suffer from lack of controls. GERD may be one cause of laryngeal signs and symptoms in a subgroup of patients but not in all those currently so suspected. Future, studies are needed in this area to better delineate this association. PMID- 15128338 TI - Asthmatics have more nocturnal gasping and reflux symptoms than nonasthmatics, and they are related to bedtime eating. AB - OBJECTIVES: Certain pulmonary diseases are now recognized as possible complications of gastroesophageal reflux (GER) disease. To further clarify the relationship between GER and asthma, we determined the prevalence, nature, and patterns of reflux symptoms in consecutive asthmatics and a well-defined patient population control group. METHODS: Two hundred and sixty-one asthmatic outpatients with well-documented asthma were interviewed in person using an extensive questionnaire. To avoid selection bias, we (a) used no selection criteria other than asthma, (b) interviewed every identified asthmatic from either the outpatient general medical clinic or pulmonary clinic (and excluded the gastroenterology clinic), and (c) excluded asthmatics referred because of gastrointestinal symptoms. A control group comprised 218 consecutive outpatients chosen from the same general medical clinics in which the asthmatics were enrolled. Interviews were conducted by one of two investigators. RESULTS: The control and asthmatic groups were similar with regard to age, gender, ethnicity, and consumption of tobacco and alcohol. There were major significant differences between the asthmatics and controls with regard to the age of onset of pulmonary and reflux symptoms, prevalence of eating before bedtime, prevalence of reflux symptoms, the quality of reflux symptoms, and the factors that promote and relieve reflux symptoms. Heartburn, regurgitation, and dysphagia were present in 71%, 45%, and 22% of asthmatics compared with 51%, 30%, and 5% of controls (p < 0.001). Three times as many asthmatics as controls had heartburn occurring throughout the day and night (OR; 95% CI: 19.5; 4.5-85.7), and three times as many asthmatics as controls had sudden nocturnal awakening with reflux symptoms and reflux-associated pulmonary symptoms that occurred simultaneously with the reflux symptoms (p < 0.001). Within the asthma group, reflux symptoms were similar in those who required and those who did not require continuous bronchodilator therapy. In these asthmatics, however, those requiring continuous bronchodilator therapy (more severe asthma) developed pulmonary and GER symptoms at a significantly older age. Eating before bedtime was recognized by significantly more asthmatics than controls as a promoter of serious nocturnal GER symptoms (4.5; 2.7-7.7). In terms of patient awareness, one-third of the asthmatics with heartburn had previously considered a relationship between their reflux symptoms and their asthma. CONCLUSION: Compared to nonasthmatics, asthmatics have significantly more frequent and more severe day and night GER symptoms and significantly more of the pulmonary symptoms (nocturnal suffocation, cough, or wheezing) so often attributed to GER. The habit of eating before bedtime appears in asthmatics to have serious and life-threatening consequences. PMID- 15128339 TI - Proton pump inhibitors and the time trends for esophageal dilation. AB - AIM: Little is known about the impact of widespread proton pump inhibitor use on the need for dilation of esophageal strictures. To determine the time trends for upper endoscopy and stricture dilation before and after the availability of proton pump inhibitors. METHODS: The computerized databases of two large community hospitals were analyzed. Annual data on dilation procedures (by all methods) and upper endoscopy were obtained and analyzed in quartiles from 1986 to 2001 to determine if the relative or absolute need for dilation changed following the introduction of proton pump inhibitors in the United States. The number of prescriptions for acid suppressive agents (H2 receptor antagonists and proton pump inhibitors) for the corresponding time period was obtained from the IMS health database. RESULTS: A total of 57,496 upper GI endoscopies and 2,868 esophageal dilations were performed. The proportion of patients undergoing esophageal dilation increased from 4% in 1986-1989 to 6% in 1990-1993 (p < 0.001), remained unchanged (5.8%) in the period from 1994 to 1997 but declined significantly to 3.9% (p < 0.001) in the next quartile (1998-2001). Absolute numbers of dilation procedures declined significantly in the last quartile (1998 2001) and the proton pump inhibitor prescriptions in the United States increased markedly from 1995 onward. CONCLUSIONS: The need for stricture dilation peaked in 1994 and has declined thereafter, corresponding to an increase in the use of proton pump inhibitors. The widespread use of proton pump inhibitors has not, however, abolished the need for esophageal dilation. PMID- 15128340 TI - Correlation between number of eosinophils and reflux index on same day esophageal biopsy and 24 hour esophageal pH monitoring. AB - OBJECTIVES: The presence of eosinophils on esophageal biopsy is a marker of esophagitis in children. Eosinophilic inflammation without evidence of gastroesophageal reflux has led to the new diagnosis of eosinophilic, or allergic, esophagitis. The aim of this study was to correlate the number of eosinophils with the reflux index on same day esophageal biopsy and 24 h esophageal pH monitoring. METHODS: A retrospective analysis of data collected from children who underwent same day endoscopy with esophageal biopsies and 24 h esophageal pH monitoring over a 3-yr period was performed. The patients were divided into five groups: Group 1: 0 eosinophils/hpf and no histologic change, Group 2: 0 eosinophils/hpf but histologic changes, Group 3: 1-5 eosinophils/hpf, Group 4: 6-20 eosinophils/hpf, and Group 5: >20 eosinophils/hpf. Reflux indices were analyzed within each group. RESULTS: A total of 305 patients met the inclusion criteria. The mean reflux indices +/- standard error within each group were Group 1 (n = 171): 2.14 +/- 0.18%, Group 2 (n = 40): 3.93 +/- 1.24%, Group 3 (n = 42): 5.96 +/- 1.53%, Group 4 (n = 21): 4.18 +/- 1.27%, and Group 5 (n = 31): 2.02 +/- 0.53%. The mean reflux index in Group 3 was significantly greater than Groups 1 and 5. CONCLUSIONS: The presence of denser infiltrates of eosinophils does not correlate with increased gastroesophageal reflux. The finding of more than 20 eosinophils/hpf is likely associated with a normal reflux index and a nonacid-related cause of esophagitis. PMID- 15128342 TI - Development of esophageal hypersensitivity following experimental duodenal acidification. AB - OBJECTIVES: As visceral afferents from different regions of the gastrointestinal tract converge at the level of the spinal cord, we hypothesized that sensitization of one gut organ would induce visceral hypersensitivity in another gut organ, remote to the sensitizing stimulus. METHODS: Protocol 1: Eight healthy male volunteers, age 30 +/- 8.2 yr, underwent three studies on different days. Esophageal pain thresholds (PT) were recorded at 10-min intervals prior to and for 2 h following a 30-min duodenal infusion of either 0.15 M hydrochloric acid (HCl), saline, or no infusion. Five subjects repeated the study to demonstrate reproducibility. Protocol 2: Esophageal evoked potentials (EEP) were studied in six subjects on two occasions prior to and 1 h after a 30-min duodenal infusion of 0.15 M HCl or saline. RESULTS: Protocol 1: After acid infusion, there were reproducible reductions in esophageal PT (ICC = 0.88), which were maximal at 110 min (15.05 +/- 2.25 mA) (p < 0.002). Following saline infusion there was an increase in esophageal PT (ICC = 0.71), which was similar to the no-infusion condition (6.21 +/- 1.54 mA vs 8.5 + 7.6 mA; p > 0.05). Protocol 2: Esophageal sensation scores increased (p= 0.02) after acid, but not after saline infusion (p= 0.1). A comparison of the latencies of EEP components prior to and following acid and saline infusion revealed a reduction in the N1 (p= 0.02) and P2 components (p= 0.04). CONCLUSION: This study provides the first objective evidence that duodenal acidification can induce esophageal hypersensitivity associated with changes in sensitivity of the central visceral pain pathway. As the esophagus was remote from the sensitizing stimulus, central sensitization of spinal dorsal horn neurons is likely to have contributed to these changes. PMID- 15128343 TI - Acid in the duodenum. AB - Acid in the duodenum triggers many physiological responses. Also, acid in the duodenum elicits hyperalgesia of the foregut. In the article by Hobson et al. in this issue the reduction of the latencies in evoked potential in response to electrical stimulation of the esophagus after duodenal acidification is reported, which may explain the symptom development in functional dyspepsia. Microscopic damages of the duodenal epithelium might be responsible for such changes. Microscopic damages of the intestinal epithelium may come into the center stage of the pathogenesis of functional GI disorders. PMID- 15128341 TI - Childhood GERD is a risk factor for GERD in adolescents and young adults. AB - OBJECTIVE: The clinical course of gastroesophageal reflux disease (GERD) in children without comorbid illness (neurological deficits, congenital esophageal anomalies, chronic obstructive airway conditions) is unclear. Whether GERD in childhood progresses or predisposes to GERD in adulthood remains unknown. METHODS: We identified a cohort of individuals endoscopically diagnosed with GERD in childhood between 1990 and 1996. We excluded patients with comorbid illnesses. Eligible persons were contacted by telephone in person or through a household member and requested to complete a validated (in adults) symptom questionnaire between February 2001 and February 2003. Respondents were invited to undergo an upper endoscopy or to share results of any endoscopic examination performed within the past 12 months. We calculated the proportion of persons with GERD symptoms (monthly, weekly), and with current use of antisecretory medications (histamine-2-receptor antagonists [H2RA], proton pump inhibitors [PPI]). RESULTS: A total of 207 persons satisfied the inclusion and exclusion criteria and were contacted. Of those, 80 (39%) completed the questionnaire and 14/80 (18%) had an upper endoscopy. The mean age of participants was 20 years (SD = 4, range 10-40); most were Caucasian (73%), and 60% were female. GERD was documented at a mean age of 5 years (approximately a 15-yr duration of follow-up). Most participants (64/80, 80%) had at least monthly heartburn and/or acid regurgitation reported within the past 12 months; 18/80 (23%) reported at least weekly symptoms, and an additional three patients were asymptomatic but taking antisecretory therapy (H2RA or PPI). If all nonresponders were considered free of symptoms, then at least 31% had monthly symptoms, and 9% had weekly symptoms. Overall, 24 (30%) were currently taking either H2RA or PPI, and 19 patients had undergone fundoplication. There were no statistically significant differences between those who reported monthly GERD symptoms, weekly GERD symptoms, or no GERD symptoms as far as demographic features, age of GERD onset, receipt of fundoplication, or current GERD treatment. At endoscopy, three patients had mild to moderate erosive esophagitis. CONCLUSIONS: GERD in otherwise normal children can persist through adolescence and adulthood in a significant proportion of patients who continue to have GERD symptoms and signs, and use antisecretory medications. Childhood GERD is a risk factor for GERD in adolescence and adulthood. PMID- 15128344 TI - Effect of proton pump inhibitors and antacid therapy on 13C urea breath tests and stool test for Helicobacter pylori infection. AB - OBJECTIVE: There is uncertainty about the best method of testing patients for Helicobacter pylori (H. pylori) infection while they are taking proton pump inhibitors. The aim of this study was to determine: (i) if the decreased sensitivity of the urea breath test during proton pump inhibitor is corrected by different techniques for breath testing and (ii) if the sensitivity of stool test is decreased with the administration of proton pump inhibitors. METHODS: Prospective randomized single-blind study was performed in a tertiary care university hospital. Out of 72 H. pylori infected patients endoscoped for upper abdominal symptoms 48 were randomized to proton pump inhibitors (omeprazole 20 mg each day or esomeprazole 40 mg each day) and 24 to antacid (aluminum hydroxide 800 mg each day) for 14 days. Several breath tests (standard 75 mg (13)C-UBT with citric acid, with orange juice, a tablet breath test with 100 and 50 mg of (13)C), and a stool test were carried out. Baseline samples were collected before and after treatment. RESULTS: The baseline sensitivity for all breath tests was 100% in both groups; for stool test it was 97.8% (95% CI: 88.7-96.6) and 90% (95% CI: 69.9-97.2) in the proton pump inhibitor and antacid group, respectively. After treatment, the sensitivity of tests was significantly low (UBTs range: 77.1%-85.4%; stool test: 83%; 95% CI: 63.9-91.1), while it was unchanged in the antacid group. CONCLUSIONS: False negative breath and stool tests are equally common in patients taking proton pump inhibitors. Antacids do not impair the sensitivity of the breath tests or the stool test. PMID- 15128345 TI - A national survey of flexible sigmoidoscopy training in primary care graduate and postgraduate education programs. AB - OBJECTIVES: Expanding the pool of primary care endoscopists to perform flexible sigmoidoscopy (FS) has been advocated as a strategy for building colorectal cancer screening capacity. The principal aim of this study was to evaluate the availability and structure of FS training among internal medicine (IM), family practice (FP), physician assistant (PA), and nurse practitioner (NP) training programs. METHODS: A postal survey of all accredited IM (n = 445), FP (n = 471), PA (n = 118), and NP (n = 149) training programs nationwide was conducted. The primary outcome was the proportion of programs offering or mandating FS training; and secondary outcomes, if applicable, were the number of participating trainees, the number of required procedures, the availability of instruction in endoscopic biopsy technique, mentors, and barriers. RESULTS: The overall response rate was 63%. Most IM (89%) and FP (99%) programs offered FS training versus only 12% of PA and 0% of NP programs. Family practice programs were more likely to offer training (p < 0.0001), require training (p < 0.0001), and teach biopsy techniques (p < 0.0001); Internal medicine programs were more likely to have minimum requirements (p < 0.0001) and required >/= 25 procedures per trainee (p < 0.0001). Physician assistant programs were less structured and often lacked minimum requirements. CONCLUSIONS: Flexible sigmoidoscopy training is widely available among FP and IM programs but more restricted or nonexistent among PA and NP programs. The lack of minimum standards for ensuring competency highlights the need for a standardized credentialing process. PMID- 15128346 TI - Colorectal cancer screening in primary care: the long and short of it. AB - Primary care physicians and physician extenders have been encouraged to perform sigmoidoscopy to improve compliance with screening and address the potential shortage of colonoscopists. A survey of primary care teaching programs reveals that sigmoidoscopy training is inconsistent and lacks minimum standards of competency. It would be helpful to know how many primary caregivers offer sigmoidoscopy in their practice and if they are deterred by insufficient resources and falling reimbursement. Colonoscopy is a more complete and efficient examination than sigmoidoscopy. Primary care physicians could be encouraged to promote colonoscopy if scheduling and patient instructions were easily available. It behooves colonoscopists to provide such assistance. PMID- 15128347 TI - Assessment of patients' perceptions of bowel preparation quality at colonoscopy. AB - AIMS: There is little published literature evaluating the accuracy of patients' perceptions of the quality of their own bowel preparation for colonoscopy. The aim of this article was to compare patients' perceptions of the adequacy of their bowel preparation with the endoscopists' rating at colonoscopy. METHODS: Outpatients undergoing elective colonoscopy completed surveys assessing bowel preparation. Patient responses regarding quality of bowel preparation were compared with endoscopists' assessment of colonic preparation. A residual stool score was also calculated for each subject based on the amount of stool, consistency of residual stool, and percentage of bowel visualized. RESULTS: A total of 474 patients were enrolled. Patients' perceptions of the quality of their bowel preparation were inaccurate when compared to the endoscopists' rating (sensitivity, 75%; specificity, 34%; accuracy, 50%). Overall correlation with endoscopists' rating was low, r = 0.08. Young patient age (<61 yr) was an independent predictor of both adequate bowel preparation ( p= 0.009) and agreement of patient/endoscopist ratings ( p= 0.003). CONCLUSIONS: Patients are unreliable judges of the quality of their own bowel preparation, tending to overestimate the cleanliness of their colon. Conversely, a patient's fear that their preparation is suboptimal is also inaccurate. A colonoscopy should not be canceled on the basis of a patient's perception that the quality of their preparation is poor. PMID- 15128348 TI - Endoscopic ultrasound-guided fine needle aspiration and multidetector spiral CT in the diagnosis of pancreatic cancer. AB - INTRODUCTION: Endoscopic ultrasound (EUS) is now established as a valuable imaging test for diagnosing and staging pancreatic cancer. But, with significant recent improvements in spiral CT scanners, particularly higher resolution and ability to reconstruct 3D images, spiral CT is now increasingly accepted as being better for pancreatic cancer staging. The debate continues, however, about the best diagnostic test or combination of tests in patients with suspected pancreatic cancer. Spiral CT is more readily available than EUS-FNA and, therefore, more frequently used. In this study, we evaluated the use of EUS-FNA in conjunction with spiral CT for suspected pancreatic cancer. METHODS: We retrospectively evaluated 81 consecutive patients who underwent EUS and EUS-FNA for clinical suspicion of a pancreatic cancer from November 2000 to November 2001. All patients had spiral CT with a multiphasic pancreatic protocol using multidetector spiral CT scanners. In all patients, EUS-FNA and spiral CT examinations were performed less than 3 wk apart. RESULTS: Overall, the accuracy of spiral CT, EUS, and EUS-FNA was 74% (n = 60/81, CI 63-83%), 94% (n = 76/81, CI 87-98%), and 88% (n = 73/81, CI 81-96%), respectively, for diagnosing pancreatic cancer. In patients without an identifiable mass on spiral CT, the diagnostic accuracy of EUS and EUS-FNA for pancreatic tumors was 92% (n = 23/25, CI 74-99%). Absence of a focal "mass" lesion on EUS reliably excluded pancreatic cancer irrespective of clinical presentation (NPV 100% n = 5/5, CI 48-100%). The negative predictive value of EUS-FNA was only 22% (n = 2/9, CI 3-60%) in patients with obstructive jaundice and biliary stricture. However, in patients without obstructive jaundice at initial presentation, EUS-FNA was highly accurate (accuracy 97%, n = 33/34, CI 85-100%) and was reliable for ruling out malignancy (NPV 89%, n = 8/9, CI 52-100%). Cytologic assessment of EUS-FNA specimens was 89% accurate for identifying malignancy in suspicious lesions visualized on EUS. CONCLUSIONS: The EUS with FNA can be a valuable adjunct to newer high-resolution multidetector spiral CT for diagnostic evaluation of patients with suspected pancreatic cancer. PMID- 15128349 TI - Miniprobe ultrasonography for determining prognosis in corrosive esophagitis. AB - OBJECTIVE: We evaluated the ability of endoscopic ultrasonography to predict likelihood of stricture formation in patients with corrosive esophagitis. METHODS: Consecutive patients with esophagitis resulting from alkaline or acid chemical ingestion (n = 11) were evaluated prospectively by endoscopic ultrasonography between hospital days 4 and 12. Findings for the most severe lesion were classified according to the appearance of the muscular layers: distinct muscular layers without thickening (grade 0); distinct muscular layers with thickening (grade I); obscured muscular layers with indistinct margins (grade II); and muscular layers that could not be differentiated (grade III). Findings were also classified according to whether apparent damage to muscular layers in the worst-appearing image involved part of the circumference (type a) or the whole circumference (type b). Implications of these findings for subsequent stricture formation were then evaluated. RESULTS: Stricture formation did not occur in patients with grade 0 or grade I images; transient stricture formation occurred in a patient showing grade IIa. Stricture requiring repeated bougie dilation occurred in a patient showing grade IIIb. CONCLUSIONS: Endoscopic ultrasonographic images presumed to reflect the destruction of muscular layers (grades II to III), as opposed to only edema (grade I), may be associated with stricture formation. This modality can accurately visualize deep lesions in corrosive esophagitis, making it prognostically useful. PMID- 15128350 TI - Lack of evidence of sexual transmission of hepatitis C among monogamous couples: results of a 10-year prospective follow-up study. AB - The risk of sexual transmission of hepatitis C virus (HCV) infection was evaluated among 895 monogamous heterosexual partners of HCV chronically infected individuals in a long-term prospective study, which provided a follow-up period of 8,060 person-years. Seven hundred and seventy-six (86.7%) spouses were followed for 10 yr, corresponding to 7,760 person-years of observation. One hundred and nineteen (13.3%) spouses (69 whose infected partners cleared the virus following treatment and 50 who ended their relationship or were lost at follow-up) contributed an additional 300 person-years. All couples denied practicing anal intercourse or sex during menstruation, as well as condom use. The average weekly rate of sexual intercourse was 1.8. Three HCV infections were observed during follow-up corresponding to an incidence rate of 0.37 per 1,000 person-years. However, the infecting HCV genotype in one spouse (2a) was different from that of the partner (1b), clearly excluding sexual transmission. The remaining two couples had concordant genotypes, but sequence analysis of the NS5b region of the HCV genome, coupled with phylogenetic analysis showed that the corresponding partners carried different viral isolates, again excluding the possibility of intraspousal transmission of HCV. Our data indicate that the risk of sexual transmission of HCV within heterosexual monogamous couples is extremely low or even null. No general recommendations for condom use seem required for individuals in monogamous partnerships with HCV-infected partners. PMID- 15128351 TI - Clinical significance of elevated alpha-fetoprotein (AFP) in patients with chronic hepatitis C, but not hepatocellular carcinoma. AB - BACKGROUND: Although elevated serum alpha-fetoprotein (AFP) is often seen in patients with chronic hepatitis C (CHC), its prevalence, risk factors, and clinical significance remain to be determined. AIMS: The present study assessed the frequency of, the risk factors for, and the clinical significance of elevated AFP in patients with CHC, but not hepatocellular carcinoma. METHODS: This retrospective study utilized systematic chart review and statistical analyses to investigate 357 U.S. patients with CHC from a university medical center and a regional veteran administration medical center. RESULTS: The prevalence of elevated serum AFP (i.e., >/=10.0 microg/L) was 23.0%, including 15.3% (28/183), 24.5% (25/102), and 42.0% (29/69) in patients with chronic hepatitis C and stage 0-II, III, and IV hepatic fibrosis, respectively. After adjusting for age, HCV load, and hepatic steatosis, stage III/IV fibrosis, elevated aspartate aminotransferase (AST), and prolonged prothrombin time as measured by international normalized ratio (INR) remained independently associated with elevated serum AFP in these patients. A serum AFP level of 15.0 microg/L was 22.8% sensitive and 94.5% specific for stage III/IV fibrosis. CONCLUSIONS: In patients with chronic hepatitis C, 23.0% had elevated serum AFP that is independently associated with stage III/IV hepatic fibrosis, elevated level of AST, and prolonged INR. PMID- 15128352 TI - Treatment of chronic hepatitis C virus in the virginia department of corrections: can compliance overcome racial differences to response? AB - OBJECTIVE: Chronic hepatitis C virus (HCV) is common in the correctional setting and there are few data on the use of interferon (IFN)/ribavirin(RVN) combination therapy in this population. Given the high proportion of African Americans (AA) in correctional facilities, which may be associated with reduced response rates, the correctional setting allows a unique opportunity to compare the response rates of AA to Caucasians (CA). The present study describes our experience of treating HCV in the inmate population of the Virginia Department of Corrections. METHODS: Of the 119 inmates evaluated between 1998 and 2000, a retrospective analysis of 59 consecutive inmates (mean age 41, 83% male, 55% CA, 73% genotype (GT)1, and 41% with advanced fibrosis) who underwent HCV therapy with IFN a-2b (3 MU TIW) and RVN (1,000-1,200 mg/d) under direct observation was performed. Patients were followed by telemedicine and the primary endpoint was sustained virologic response (SVR) defined as an undetectable HCV RNA at least 24 wk after completion of therapy. RESULTS: All but one patient completed at least 12 wk of therapy and no patient required dose reduction. By wk 24, 34 inmates (58%) responded (negative HCV RNA) which was higher in CA compared to AA (70%vs 40%; p= 0.037). Although overall SVR was higher in CA compared to AA (41%vs 28%; p= ns), we observed no difference in SVR when comparing only GT 1 CA to AA (33%vs 29%). CONCLUSIONS: HCV can be effectively treated in the correctional setting with response rates similar to, if not better than the published literature. In this controlled setting of direct observational therapy, we observed similar SVR in CA and AA. PMID- 15128353 TI - Sudden hearing loss in patients with chronic hepatitis C treated with pegylated interferon/ribavirin. AB - BACKGROUND: Sudden hearing loss has been reported on standard interferon (IFN) alpha2 therapy. This is the first report on the occurrence of sudden hearing loss in six cases of chronic hepatitis C in temporal relation to treatment with pegylated (PEG)-IFN alfa2a or b/ribavirin combination therapy. Three patients were treated in an ongoing randomized placebo-controlled trial comparing the addition of 200 mg amantadine or placebo to the combination of 180 microg PEG-IFN alpha2a (PEGASYS, Roche, Basel, CH)/wk and 1-1.2 g ribavirin/d (COPEGUS, Roche, Nutley, USA) in de novo patients infected with HCV genotype 1. Sudden hearing loss and tinnitus developed on day 1 and after 4, 23, 25, 36, and 40 wk of treatment, respectively. CONCLUSIONS: Sudden hearing loss may occur in about 1% of patients on PEG-IFN/ribavirin combination therapy. This rate was not different to that observed in an untreated population. Possible mechanisms involved include direct ototoxicity of IFN, autoimmunity, and hematological changes. In contrast to published cases on auditory disability due to standard IFN, hearing loss did not fully resolve after discontinuation of therapy with PEG-IFN. On the other hand, symptoms did not worsen on continued treatment. Therefore, the decision whether to continue or to stop the treatment when signs of ototoxicity appear is based on the clinical judgment of the treating physician. PMID- 15128354 TI - Early postoperative complications are not increased in patients with Crohn's disease treated perioperatively with infliximab or immunosuppressive therapy. AB - AIM: The aim was to determine whether the use of steroids, immunosuppressive agents, or infliximab prior to abdominal surgery for Crohn's disease is associated with an increased rate of early postoperative complications. METHODS: All patients who underwent abdominal surgery for Crohn's disease between October 1998 and December 2001 were identified. Medical records were abstracted for demographics, location and duration of disease, use of infliximab within 8 wk before and 4 wk after surgery, and dose and duration of corticosteroids, azathioprine/6-mercaptopurine, and methotrexate. Steroid use was defined as: high (intravenous or oral >/=40 mg/day), moderate (oral >/=20 mg/day for at least 2 months), low (oral <20 mg/day or oral >20 mg/day for <2 months), or none. Early (within 30 days postinfliximab) septic and nonseptic complications were identified. Septic complications included wound sepsis, intraabdominal, and extraabdominal infections. Nonseptic complications included Crohn's disease recurrence, small bowel obstruction, gastrointestinal bleeding, and thromboembolism. A logistic regression analysis assessed the association between perioperative therapy with infliximab, corticosteroids, or immunosuppressive therapy and subsequent occurrence of septic complications and separately overall complications. RESULTS: Two hundred and seventy patients were operated upon including 107 patients who received steroids (34 low dose, 34 moderate dose, 43 high dose), 105 patients who received immunosuppressives (64 azathioprine, 38 6 mercaptopurine, 4 methotrexate), and 52 who received infliximab. Forty-eight patients underwent urgent or emergent surgery and 222 underwent elective surgery. Septic complications occurred in 52 of 270 (19%) patients including wound sepsis in 28 (10%), anastomotic leak in 9 (3%), intraabdominal abscess in 5 (2%), and extraabdominal infections in 19 (7%). Nonseptic complications occurred in 18 of 270 (7%) patients. Preoperative use of high- or moderate-dose steroids, immunosuppressives, or infliximab was not associated with greater complication rates. No deaths occurred. CONCLUSION: Early complications after elective abdominal surgery for CD are not associated with steroid dose, immunosuppressive therapy, or infliximab use. PMID- 15128355 TI - Inflammatory bowel diseases, 5-aminosalicylic acid and sulfasalazine treatment and risk of acute pancreatitis: a population-based case-control study. AB - OBJECTIVES: Patients with inflammatory bowel diseases are suggested to have an increased risk of acute pancreatitis. Although azathioprine and glucocorticoids are risk factors for acute pancreatitis, the relation is poorly understood, in particular the role of 5-aminosalicylic acid and sulfasalazine treatment. To clarify these relations, we conducted a population-based case-control study. METHODS: We identified 1,590 incident cases of acute pancreatitis from the Hospital Discharge Registry of the North Jutland County of Denmark from 1991 to 2002, and selected 10 controls per case (N = 15,913) from the Central Personal Registry, matched by age and gender. Among cases and controls, we identified patients with inflammatory bowel diseases. Data on drug use were extracted from a Pharmaco-epidemiological Prescription Database. RESULTS: Adjusted odds ratios for acute pancreatitis in patients with Crohn's disease and ulcerative colitis were 3.7 (95% confidence interval (CI), 1.9-7.6) and 1.5 (95% CI, 0.7-3.6), respectively. In all patients treated with 5-aminosalicylic acid and sulfasalazine the adjusted odds ratios for acute pancreatitis were 0.7 (95% CI, 0.4-2.2) and 1.5 (95% CI, 0.4-5.2), respectively. Restricted to patients with inflammatory bowel diseases only, the adjusted odds ratios for acute pancreatitis in patients exposed to 5-aminosalicylic acid and sulfasalazine were 0.7 (95% CI, 0.1-3.8) and 0.6 (95% CI, 0.1-6.7), respectively. CONCLUSION: We found a nearly four-fold increased risk of acute pancreatitis in patients with Crohn's disease and a 1.5-fold increased risk for ulcerative colitis. In patients with inflammatory bowel diseases, the use of 5-aminosalicylic acid or sulfasalazine was not associated with increased risk of acute pancreatitis. PMID- 15128356 TI - Predictors of alternative and complementary medicine use in inflammatory bowel disease: do measures of conventional health care utilization relate to use? AB - OBJECTIVE: Alternative and complementary therapies (ACM) have gained increasing attention in the past few years. It was our purpose to determine whether increased ACM use is associated with increased use of conventional health care resources. Additionally, demographics of use, subjective benefit, and cost were analyzed. METHODS: We enrolled 150 inflammatory bowel disease (IBD) patients from a tertiary care center and performed a phone survey of their ACM use in the past year. A population-based administrative database was accessed to extract data regarding use of conventional medicine (hospitalizations, doctor visits, and GI specific doctor visits). Patients were divided into three groups: (i) no ACM (n = 60) (ii) users of exercise, diet, and prayer (EDP) exclusively (n = 47) (iii) other ACM use (n = 43) which included those who may have used EDP as well as any of acupuncture, chiropractic, homeopathy, naturopathy, herbology, massage, relaxation, reflexology, hypnotherapy, aromatherapy, meditation, or support group. RESULTS: ACM was used by 60% (EDP 31%, other ACM 29%). There were no significant differences in use between the three groups by disease diagnosis, education level, employment status, use of IBD medications, number of hospitalizations, doctor visits, or GI specific doctor visits. The EDP group was more likely to be married (p = 0.006) and female (p = 0.04) compared to no ACM. The EDP group tended to be older than the no ACM (p = 0.001) and other ACM (p = 0.01). The other ACM had shorter disease duration than EDP (p = 0.04) and no ACM (p = 0.04). The most commonly used therapies were diet (45%), herbal (17%), exercise (15%), prayer (11%), and relaxation (10%). ACM was sought for pain/cramps (64%), diarrhea (60%), and gas/bloating (21%). Seventy-three percent of EDP interventions incurred no cost compared to 33% with other ACM (p < 0.0001). The median annual amount spent on other ACM was $56 (range $0-$4800). Subjectively, patients felt helped by trials of EDP 95% of the time whereas other ACM helped 67% of the time (p < 0.0001). CONCLUSIONS: ACM use could not be predicted by either greater or less hospitalizations, conventional doctor visits, or GI specific visits. ACM was sought mostly to palliate pain or diarrhea. Those using EDP are more likely to be older married women. Subjectively other ACM is of less benefit (67%) than EDP (95%). If doctor visits or hospitalizations represent degree of increased disease activity then this too is not predictive of using ACM. PMID- 15128357 TI - Presence of bacteria and innate immunity of intestinal epithelium in childhood celiac disease. AB - OBJECTIVES: Exposure to gliadin and related prolamins and appropriate HLA-DQ haplotype are necessary but not sufficient for contracting celiac disease (CD). Aberrant innate immune reactions could be contributing risk factors. Therefore, jejunal biopsies were screened for bacteria and the innate immune status of the epithelium investigated. METHODS: Children with untreated, treated, challenged CD, and controls were analyzed. Bacteria were identified by scanning electron microscopy. Glycocalyx composition and mucin and antimicrobial peptide production were studied by quantitative RT-PCR, antibody and lectin immunohistochemistry. RESULTS: Rod-shaped bacteria were frequently associated with the mucosa of CD patients, with both active and inactive disease, but not with controls. The lectin Ulex europaeus agglutinin I (UEAI) stained goblet cells in the mucosa of all CD patients but not of controls. The lectin peanut agglutinin (PNA) stained glycocalyx of controls but not of CD patients. mRNA levels of mucin-2 (MUC2), alpha-defensins HD-5 and HD-6, and lysozyme were significantly increased in active CD and returned to normal in treated CD. Their expression levels correlated to the interferon-gamma mRNA levels in intraepithelial lymphocytes. MUC2, HD-5, and lysozyme proteins were seen in absorptive epithelial cells. beta defensins hBD-1 and hBD-2, carcinoembryonic antigen (CEA), CEA cell adhesion molecule-1a (CEACAM1a), and MUC3 were not affected. CONCLUSIONS: Unique carbohydrate structures of the glycocalyx/mucous layer are likely discriminating features of CD patients. These glycosylation differences could facilitate bacterial adhesion. Ectopic production of MUC2, HD-5, and lysozyme in active CD is compatible with goblet and Paneth cell metaplasia induced by high interferon gamma production by intraepithelial lymphocytes. PMID- 15128358 TI - A role for bacteria in celiac disease? AB - The finding of rod-shaped bacteria attached to the small intestinal epithelium of some untreated and treated celiac-disease patients, but not to the epithelium of healthy controls, ignites the notion that bacteria may be involved in the pathogenesis of celiac disease. This editorial discusses this possibility in relation to the current understanding of the molecular basis of this disorder. PMID- 15128359 TI - Critical factors affecting quality of life of adult patients with anorectal malformations or Hirschsprung's disease. AB - OBJECTIVES: The first objective was to compare the quality of life of adult patients with anorectal malformations (ARM) or Hirschsprung's disease (HD) and to compare both groups with healthy people. And the second objective was to examine the factors that affect the quality of life of patients with ARM or HD, using a theoretical model in which patients' background characteristics explain quality of life via mediating disease-specific functioning and psychosocial functioning. METHODS: Three hundred and forty-one patients completed a questionnaire, which assessed sociodemographic characteristics, disease-specific and psychosocial functioning, and quality of life. Clinical factors were extracted from the medical records. RESULTS: Patients with ARM or Hirschsprung's disease did not differ in their quality of life. Compared to healthy people, both patient groups reported more limitations in their "overall" physical quality of life, but only patients with ARM reported impaired quality of life on several specific domains (e.g., physical role-functioning, pain). The model was largely accepted. Most striking were the strong effects of the psychosocial functioning factors in contrast to weak effects of the disease-specific "constipation" and "fecal continence" factors. CONCLUSIONS: The quality of life of patients with ARM or Hirschsprung's disease was found to be comparable. Compared to healthy people, both patient groups encountered "overall" physical health problems, but only patients with ARM reported additional pain and limitations in role functioning due to physical problems. It appeared that psychosocial functioning had the most important effect on the quality of life of patients with ARM or Hirschsprung's disease, while fecal incontinence and constipation had almost no effect on their quality of life. PMID- 15128360 TI - Paroxetine to treat irritable bowel syndrome not responding to high-fiber diet: a double-blind, placebo-controlled trial. AB - OBJECTIVES: The purpose of the trial was to determine whether a high-fiber diet (HFD) alone or in combination with paroxetine or placebo was effective treatment for patients with irritable bowel syndrome (IBS). METHODS: DESIGN: Trial of HFD alone (Group 1) followed by a randomized, double-blind trial of HFD with paroxetine or placebo (Group 2). SETTING: Gastroenterology office in a 524-bed university-affiliated community hospital in Pittsburgh. PATIENTS: Men and women, aged 18-65 yr, previously diagnosed with IBS but otherwise healthy. INTERVENTION: Institution of HFD in 98 participants consuming low- or average-fiber diets. Allocation of paroxetine to 38 and placebo to 43 symptomatic participants consuming HFDs. MEASUREMENTS: Overall well-being, abdominal pain, and abdominal bloating (Groups 1 and 2); food avoidance, work functioning, and social functioning (Group 2). RESULTS: In Group 1, overall well-being improved in 26% patients, and abdominal pain and bloating decreased in 22% and 26% patients, respectively, with an HFD. In Group 2, overall well-being improved more with paroxetine than with placebo (63.3%vs 26.3%; p= 0.01), but abdominal pain, bloating, and social functioning did not. With paroxetine, food avoidance decreased (p= 0.03) and work functioning was marginally better (p= 0.08). Before unblinding, more paroxetine recipients than placebo recipients wanted to continue their study medication (84%vs 37%; p < 0.001). CONCLUSIONS: The difference in overall well-being found in our paroxetine/placebo trial is greater than that found in previously published drug/placebo trials for IBS. Moreover, the difference in well-being applied to nondepressed recipients of paroxetine. PMID- 15128361 TI - Antidepressants in IBS: are we deluding ourselves? AB - The benefit of selective serotonin reuptake inhibitors (SSRIs) in the irritable bowel syndrome (IBS) has not been clear. In the latest randomized trial published this month in the Journal, paroxetine was superior to placebo in terms of improving well-being, but not abdominal pain or bloating. Based on the results of the most recent studies, both tricyclic antidepressants and SSRIs may improve patient satisfaction or quality-of-life without relieving most of the primary gastrointestinal symptoms. This suggests that antidepressant therapy represents at best only a "band-aid" approach to management. Optimizing the use of antidepressants in IBS is a challenge, and these issues are explored in this Editorial. PMID- 15128362 TI - The prevalence, symptom characteristics, and impact of irritable bowel syndrome in an asian urban community. AB - OBJECTIVES: To study the epidemiology, symptom characteristics and impact of IBS in an urban Asian population. METHODS: A validated bowel symptom questionnaire was administered at face-to-face interviews to a random sample of 3,000 households in Singapore. RESULTS: The response rate was 78.2% (n = 2,276, 1,143 males and 1,133 females). The age, sex, and racial distribution of our respondents were similar to the general population and there was no significant difference between respondents and nonrespondents by type of household. The prevalence of IBS was 11.0%, 10.4%, and 8.6% by Manning (>1 criteria), Rome I and Rome II criteria, respectively. There was a higher prevalence of IBS in those <50 years of age (9.7%vs 5.8% 50 or > years, p = 0.002), with more than 6 years of education (9.8%vs 5.9% 6 year or <, p = 0.002) and living in landed property (16.8%vs 8.2% living in apartments and public housing, p = 0.008). There was no striking preponderance of female IBS subjects. Chronic constipation was a more common bowel disturbance than chronic diarrhea among our IBS subjects (51.0%vs 12.8%). CONCLUSIONS: The prevalence and impact of IBS in our Asian urban society are greater than previously appreciated. We believe that our results provide a useful window to the future trends of gastrointestinal diseases for health and education authorities in developing Asian countries to look into. PMID- 15128363 TI - Recurrent attacks of autoimmune pancreatitis result in pancreatic stone formation. AB - OBJECTIVES: Autoimmune pancreatitis has been characterized by irregular narrowing of the main pancreatic duct and sonolucent swelling of the parenchyma, both of which are due to lymphoplasmacytic inflammation at the active stage of the disease, and by the absence of pancreatic stone formation. The aim of the present study was to confirm or deny whether or not this disease is progressive with recurrent attacks, resulting in pancreatic stone formation like ordinary chronic pancreatitis. METHODS: Forty-two patients, 36 of whom were treated with prednisolone, were followed up for periods longer than 12 months (median follow up period: 54.5 months, range: 13-111 months) by regular interview and examination of their medical records for laboratory tests and image tests. RESULTS: Eleven patients (26.2%) who were treated with prednisolone showed recurrent attacks during median follow-up periods of 22 months. Eight patients (19%) showed the formation of pancreatic stones during the follow-up periods. Because 6 of 11 patients (54.5%) who suffered relapse showed pancreatic stone formation, it is significantly associated with relapse in comparison with nonrelapse (p= 0.0019). CONCLUSIONS: Contrary to previous reports, we observed both relapse and pancreatic stone formation in some patients with autoimmune pancreatitis, which suggests that autoimmune pancreatitis has the potential to be a progressive disease with pancreatic stones. PMID- 15128364 TI - Platelets in inflammatory bowel disease: clinical, pathogenic, and therapeutic implications. AB - Both Crohn's disease (CD) and ulcerative colitis (UC) are associated with abnormalities of platelet number and function. In the peripheral circulation the state of platelet activation is typically increased, and inflammatory bowel disease (IBD)-involved mucosa frequently contains platelet aggregates within mucosal microthrombi. The relevance of platelet dysfunction to IBD pathogenesis is still unclear, but there is solid evidence demonstrating that platelets, in addition to their traditional role in hemostasis, can also function as potent proinflammatory cells. Upon activation, platelets secrete a large number of biologically active molecules able to induce or amplify an inflammatory process through many of the same cellular and molecular pathways conventionally utilized by immune cells mediating IBD. The aim of this article is to review data on the existence of platelet dysfunction in IBD, substantiate platelets' inflammatory potential, discuss the implications of abnormal platelet activity for chronic intestinal inflammation, and consider the potential benefits of platelet modulation for treatment of IBD. PMID- 15128365 TI - Gastroesophageal reflux disease: a typical spectrum disease (a new conceptual framework is not needed). AB - Gastroesophageal reflux disease (GERD) is a common GI disorder, particularly frequent in the primary care setting, with a high direct and indirect economic burden on society. Despite the high prevalence and costs of the disease, the epidemiology and natural history of GERD have not been fully elucidated. It has recently been suggested to abandon the current model of GERD as a "spectrum" disease and to adopt a new conceptual framework, e.g., categorizing GERD into three unique groups of patients: nonerosive reflux disease, erosive esophagitis, and Barrett's esophagus. In the present review we present arguments against this proposal, and argue that the concept of a single disease, potentially progressing from mild nonerosive forms toward metaplasia and neoplasia (adenocarcinoma), still holds true and may in fact help us in planning the diagnostic and therapeutic approach as well as in allocating financial resources much better than the proposed model of a "tripartited" disease. Independently from the conceptual model adopted, however, more data on the natural history of patients with GERD are eagerly needed. PMID- 15128366 TI - Fulminant liver failure due to usnic acid for weight loss. AB - The use of complementary and alternative medicine (CAM) in developed countries has increased significantly over the years. Among the most popular are the weight loss supplements or "fat burners." Liver failure due to these popular remedies has been widely recognized. Usnic acid has been an ingredient of dietary supplements that cause liver failure. Its hepatotoxicity has not been recognized because it is usually mixed with other ingredients that are presumably hepatotoxic. We describe a case of a 28-yr-old woman who presented with fulminant liver failure requiring orthotopic liver transplantation, after taking pure usnic acid for weight loss. This is the first report on fulminant liver failure associated with the ingestion of pure usnic acid. A discussion about hepatotoxicity of the different compounds of dietary supplements is presented. This is a reminder for the clinicians about the potential side effects of CAM. PMID- 15128367 TI - Utility of endoscopic ultrasound for restaging rectal cancer following neoadjuvant chemoradiation therapy. PMID- 15128369 TI - Anti-Saccharomyces cerevisiae antibody and 5-aminosalicylic acid treatment. PMID- 15128371 TI - Acute west nile virus in two patients receiving interferon and ribavirin for chronic hepatitis C. PMID- 15128372 TI - Specificity of IEL profiling in the diagnosis of celiac disease. PMID- 15128373 TI - Non-alcohol induced steatohepatitis in non-obese patients: treatment with ursodeoxycholic acid. PMID- 15128374 TI - Gut flora and the irritable bowel syndrome. PMID- 15128375 TI - Re: Scoglio et al. Is intestinal biopsy always needed for diagnosis of celiac disease? PMID- 15128376 TI - Gastrointestinal bleeding, cryoglobulinemia, and hepatitis C. PMID- 15128377 TI - Introduction to concepts and measurement of the emergence of tobacco dependence: The Tobacco Etiology Research Network. PMID- 15128378 TI - Measuring the emergence of tobacco dependence: the contribution of negative reinforcement models. AB - This review of negative reinforcement models of drug dependence is part of a series that takes the position that a complete understanding of current concepts of dependence will facilitate the development of reliable and valid measures of the emergence of tobacco dependence. Other reviews within the series consider models that emphasize positive reinforcement and social learning/cognitive models. This review summarizes negative reinforcement in general and then presents four current negative reinforcement models that emphasize withdrawal, classical conditioning, self-medication and opponent-processes. For each model, the paper outlines central aspects of dependence, conceptualization of dependence development and influences that the model might have on current and future measures of dependence. Understanding how drug dependence develops will be an important part of future successful tobacco dependence measurement, prevention and treatment strategies. PMID- 15128379 TI - Measures and models of nicotine dependence: positive reinforcement. AB - This paper addresses the problem of assessing nicotine dependence. The main objective is to develop theory-led suggestions for measures that will be relevant in the early phases of tobacco use, as well as in established smokers. Theoretical models of addiction falling into the general class of 'positive reinforcement theories' were identified and reviewed. From this review a number of drug effects and patterns of behaviour were distilled and categorized as either vulnerability or dependence indicators. A comparison of those features with the International Classification of Diseases (ICD-10) and Diagnostic and Statistical Manual (DSM-IV) diagnostic systems shows that neither system includes detailed assessment of vulnerability indicators. It is argued that measurement of vulnerability indicators, in addition to dependence indicators, may add to the predictive validity of assessments carried out in early career tobacco users, especially where there is limited evidence of established dependence. In addition, it is suggested that examination of measures that differentiate a subgroup of early career smokers termed 'rapid accelerators' may prove profitable and enable identification of the key parameters of nicotine reinforcement. PMID- 15128380 TI - Cognitive and social learning models of drug dependence: implications for the assessment of tobacco dependence in adolescents. AB - This paper is part of a series that has the goal of identifying potential approaches toward developing new instruments for assessing tobacco dependence among adolescents. The fundamental assumption underlying the series is that contemporary theories of drug dependence offer a rich source of opportunities for the development of theoretically based assessment tools. The present paper focuses on cognitive and social-learning models of drug dependence and the implications of these models for novel assessment instruments. In particular, the paper focuses on Mark Goldman's model of drug expectancies, Albert Bandura's model of self-efficacy, Thomas Wills's model of stress and coping and Stephen Tiffany's cognitive-processing model of drug urges and cravings. In addition to traditional self-report measures, naturalistic and laboratory-based assessments are identified that may yield information relevant to multi-dimensional measurement of tobacco dependence. PMID- 15128381 TI - What can dependence theories tell us about assessing the emergence of tobacco dependence? AB - Little is known about the processes that underlie changes in smoking that occur between the first use of a cigarette, subsequent regular use and eventual addictive use. At present, assessments of those critical processes are poorly developed and not strongly informed by contemporary models of drug dependence. The preceding three papers in this special issue address explicitly how modern drug-dependence theories describe the emergence of drug dependence and the implications of those theories for assessment. The papers covered three domains of theories: negative reinforcement, positive reinforcement, and cognitive and social learning. In this paper, we summarize these reviews and extract general themes and issues that emerge across all the articles. These include: (1) the importance of learning processes; (2) limitations of self-report measures; (3) the view of dependence as a process and not a state; (4) the conception of dependence on a continuum in contrast to the conventional perspective of tobacco dependence as a natural category; (5) the ontological status of the dependence concept; (6) limitations of backward extrapolations from adult assessments; (7) the possibility of multiple dimensions or forms of dependence; and (8) the value of a transdisciplinary approach when studying the emergence of tobacco dependence. PMID- 15128382 TI - The European Study of the Epidemiology of Mental Disorders (ESEMeD) project: an epidemiological basis for informing mental health policies in Europe. PMID- 15128383 TI - Sampling and methods of the European Study of the Epidemiology of Mental Disorders (ESEMeD) project. AB - OBJECTIVE: The European Study of Epidemiology of Mental Disorders (ESEMeD) project was designed to evaluate the prevalence, the impact and the treatment patterns in Europe. This paper presents an overview of the methods implemented in the project. METHOD: ESEMeD is a cross-sectional study in a representative sample of 21 425 adults, 18 or older, from the general population of Belgium, France, Germany, Italy, the Netherlands and Spain. The Composite International Diagnostic Interview (WMH-CIDI) was administered by home interviews from January 2001 to August 2003 using Computer Assisted Personal Interview (CAPI) technology. Data quality was controlled to ensure reliability and validity of the information obtained. RESULTS: Response rate varied from 78.6% in Spain to 45.9% in France. Less than 4% of the individuals had errors in the checking procedures performed. CONCLUSION: The sampling methodologies, comprehensive psychiatric instruments and quality control procedures used have rendered the ESEMeD database a unique and important source of information about the prevalence, the disability burden and unmet medical needs of mental disorders within Europe. PMID- 15128384 TI - Prevalence of mental disorders in Europe: results from the European Study of the Epidemiology of Mental Disorders (ESEMeD) project. AB - OBJECTIVE: To describe the 12-month and lifetime prevalence rates of mood, anxiety and alcohol disorders in six European countries. METHOD: A representative random sample of non-institutionalized inhabitants from Belgium, France, Germany, Italy, the Netherlands and Spain aged 18 or older (n = 21425) were interviewed between January 2001 and August 2003. DSM-IV disorders were assessed by lay interviewers using a revised version of the Composite International Diagnostic Interview (WMH-CIDI). RESULTS: Fourteen per cent reported a lifetime history of any mood disorder, 13.6% any anxiety disorder and 5.2% a lifetime history of any alcohol disorder. More than 6% reported any anxiety disorder, 4.2% any mood disorder, and 1.0% any alcohol disorder in the last year. Major depression and specific phobia were the most common single mental disorders. Women were twice as likely to suffer 12-month mood and anxiety disorders as men, while men were more likely to suffer alcohol abuse disorders. CONCLUSION: ESEMeD is the first study to highlight the magnitude of mental disorders in the six European countries studied. Mental disorders were frequent, more common in female, unemployed, disabled persons, or persons who were never married or previously married. Younger persons were also more likely to have mental disorders, indicating an early age of onset for mood, anxiety and alcohol disorders. PMID- 15128385 TI - 12-Month comorbidity patterns and associated factors in Europe: results from the European Study of the Epidemiology of Mental Disorders (ESEMeD) project. AB - OBJECTIVE: Comorbidity patterns of 12-month mood, anxiety and alcohol disorders and socio-demographic factors associated with comorbidity were studied among the general population of six European countries. METHOD: Data were derived from the European Study of the Epidemiology of Mental Disorders (ESEMeD), a cross sectional psychiatric epidemiological study in a representative sample of adults aged 18 years or older in Belgium, France, Germany, Italy, the Netherlands and Spain. The diagnostic instrument used was the Composite International Diagnostic Interview (WMH-CIDI). Data are based on 21 425 completed interviews. RESULTS: In general, high associations were found within the separate anxiety disorders and between mood and anxiety disorders. Lowest comorbidity associations were found for specific phobia and alcohol abuse-the disorders with the least functional disabilities. Comorbidity patterns were consistent cross-nationally. Associated factors for comorbidity of mood and anxiety disorders were female gender, younger age, lower educational level, higher degree of urbanicity, not living with a partner and unemployment. Only younger people were at greater risk for comorbidity of alcohol disorder with mood, anxiety disorders or both. CONCLUSION: High levels of comorbidity are found in the general population. Comorbidity is more common in specific groups. To reduce psychiatric burden, early intervention in populations with a primary disorder is important to prevent comorbidity. PMID- 15128386 TI - Disability and quality of life impact of mental disorders in Europe: results from the European Study of the Epidemiology of Mental Disorders (ESEMeD) project. AB - OBJECTIVE: This manuscript examines the impact of mental health state and specific mental and physical disorders on work role disability and quality of life in six European countries. METHOD: The ESEMeD study was conducted in: Belgium, France, Germany, Italy, the Netherlands and Spain. Individuals aged 18 years and over who were not institutionalized were eligible for an in-home computer-assisted interview. Common mental disorders, work loss days (WLD) in the past month and quality of life (QoL) were assessed, using the WMH-2000 version of the CIDI, the WHODAS-II, and the mental and physical component scores (MCS, PCS) of the 12-item short form, respectively. The presence of five chronic physical disorders: arthritis, heart disease, lung disease, diabetes and neurological disease was also assessed. Multivariate regression techniques were used to identify the independent association of mental and physical disorders while controlling for gender, age and country. RESULTS: In each country, WLD and loss of QoL increased with the number of disorders. Most mental disorders had approximately 1.0 SD-unit lower mean MCS and lost three to four times more work days, compared with people without any 12-month mental disorder. The 10 disorders with the highest independent impact on WLD were: neurological disease, panic disorder, PTSD, major depressive episode, dysthymia, specific phobia, social phobia, arthritis, agoraphobia and heart disease. The impact of mental vs. physical disorders on QoL was specific, with mental disorders impacting more on MCS and physical disorders more on PCS. Compared to physical disorders, mental disorders had generally stronger 'cross-domain' effects. CONCLUSION: The results suggest that mental disorders are important determinants of work role disability and quality of life, often outnumbering the impact of common chronic physical disorders. PMID- 15128387 TI - Use of mental health services in Europe: results from the European Study of the Epidemiology of Mental Disorders (ESEMeD) project. AB - OBJECTIVE: Comprehensive information about access and patterns of use of mental health services in Europe is lacking. We present the first results of the use of health services for mental disorders in six European countries as part of the ESEMeD project. METHOD: The study was conducted in: Belgium, France, Germany, Italy, the Netherlands and Spain. Individuals aged 18 years and over who were not institutionalized were eligible for an computer-assisted interview done at home. The 21 425 participants were asked to report how frequently they consulted formal health services due to their emotions or mental health, the type of professional they consulted and the treatment they received as a result of their consultation in the previous year. RESULTS: An average of 6.4% of the total sample had consulted formal health services in the previous 12 months. Of the participants with a 12-month mental disorder, 25.7% had consulted a formal health service during that period. This proportion was higher for individuals with a mood disorder (36.5%, 95% CI 32.5-40.5) than for those with anxiety disorders (26.1%, 95% CI 23.1-29.1). Among individuals with a 12-month mental disorder who had contacted the health services 12 months previously, approximately two-thirds had contacted a mental health professional. Among those with a 12-month mental disorder consulting formal health services, 21.2% received no treatment. CONCLUSION: The ESEMeD results suggest that the use of health services is limited among individuals with mental disorders in the European countries studied. The factors associated with this limited access and their implications deserve further research. PMID- 15128388 TI - Psychotropic drug utilization in Europe: results from the European Study of the Epidemiology of Mental Disorders (ESEMeD) project. AB - OBJECTIVE: To assess psychotropic drug utilization in the general population of six European countries, and the pattern of use in individuals with different DSM IV diagnoses of 12-month mental disorders. METHOD: Data were derived from the European Study of the Epidemiology of Mental Disorders (ESEMeD/MHEDEA 2000), a cross-sectional psychiatric epidemiological study in a representative sample of 21 425 adults aged 18 or older from six European countries (e.g. Belgium, France, Germany, Italy, the Netherlands and Spain). Individuals were asked about any psychotropic drug use in the past 12 months, even if they used the drug(s) just once. A colour booklet containing high-quality pictures of psychotropic drugs commonly used to treat mental disorders was provided to help respondents recall drug use. RESULTS: Psychotropic drug utilization is generally low in individuals with any 12-month mental disorder (32.6%). The extent of psychotropic drug utilization varied according to the specific DSM-IV diagnosis. Among individuals with a 12-month diagnosis of pure major depression, only 21.2% had received any antidepressants within the same period; the exclusive use of antidepressants was even lower (4.6%), while more individuals took only anxiolytics (18.4%). CONCLUSION: These data question the appropriateness of current pharmacological treatments, particularly for major depression, in which under-treatment is coupled with the high use of non-specific medications, such as anxiolytics. PMID- 15128389 TI - Humanin antagonists: mutants that interfere with dimerization inhibit neuroprotection by Humanin. AB - The 24-residue peptide Humanin (HN) protects neuronal cells from insults of various Alzheimer's disease (AD) genes and Abeta by forming a homodimer. We have previously shown that P3A, S7A, C8A, L9A, L12A, T13A, S14A and P19A mutations nullify the neuroprotective function of HN [Yamagishi, Y., Hashimoto, Y., Niikura, T. & Nishimoto, I. (2003) Peptides, 24, 585-595]. Here we examined whether any of these 'null' mutants could function as dominant-negative mutants. Homodimerization-defective mutants, P3A-, L12A-, S14A- and P19A-HN, specifically blocked neuroprotection by HN, but not by activity-dependent neurotrophic factor. Furthermore, insertion of S7A, the mutation that blocks the homodimerization of HN, but not insertion of G5A abolished the antagonizing function of L12A-HN. While L12A-HN and G5A/L12A-HN actually inhibited HN homodimerization, S7A/L12A-HN had no effect. These data indicate that P3A-, L12A-, S14A- and P19A-HN function as HN antagonists by forming an inactive dimer with HN. This study provides a novel insight into the understanding of the in vivo function of HN, as well as into the development of clinically applicable HN neutralizers. PMID- 15128390 TI - Glycine cleavage system in neurogenic regions. AB - The glycine cleavage system (GCS) is the essential enzyme complex for degrading glycine and supplying 5,10-methylenetetrahydrofolate for DNA synthesis. Inherited deficiency of this system causes nonketotic hyperglycinemia, characterized by severe neurological symptoms and frequent association of brain malformations. Although high levels of glycine have been considered to cause the above-mentioned problems, the detailed pathogenesis of this disease is still unknown. Here we show that GCS is abundantly expressed in rat embryonic neural stem/progenitor cells in the neuroepithelium, and this expression is transmitted to the radial glia-astrocyte lineage, with prominence in postnatal neurogenic regions. These data indicate that GCS plays important roles in neurogenesis, and suggest that disturbance of neurogenesis induced by deficiency of GCS may be the main pathogenesis of nonketotic hyperglycinemia. PMID- 15128391 TI - Ligand-gated channels in early mesencephalic neuronal precursors: immunocytochemical and electrophysiological analysis. AB - Neuronal precursors play an important role in potential regenerative therapeutic strategies in different neurodegenerative diseases, e.g. Parkinson's disease. To understand proliferation and differentiation of these cells in vitro and in vivo, it is important to characterize functional properties of neuronal precursors in detail. The aim of the present study was to analyse the electrophysiological characteristics of ligand-gated channels of neuronal precursors prepared from the rat ventral mesencephalon (VM) of embryonic stage 12.5 during their in vitro differentiation. For the experiments we used the patch-clamp technique in combination with a system for ultrafast solution exchange and immunocytochemistry. It could be shown that functional active AMPA-type glutamate as well as GABA(A) receptor channels are expressed at an early stage of neuronal development. In culture we observed excitatory as well as inhibitory postsynaptic currents (defined by their different kinetics) which correspond to the activation of AMPAergic and GABAergic receptor channels. Two populations of glutamate activated currents could be differentiated by their different time course of desensitization whereas the time course of resensitization and deactivation was normally distributed in all cells. GABAergic currents could be blocked by bicuculline and their kinetics correspond to that of GABA(A) receptor channel currents. Summarizing the results, in the present study it was shown for the first time that neuronal embryonic precursors of the rat VM express both functional AMPA-type glutamate and functional GABA(A) receptor channels in vitro. PMID- 15128392 TI - Induction of neurogenesis in the adult rat subventricular zone and neostriatum following dopamine D3 receptor stimulation. AB - Discrete regions of the adult CNS, including the subventricular zone (SVZ), do retain the capacity for neurogenesis. These progenitor cells may represent a potential new source of cells for replacement therapies in neuroregenerative diseases. An understanding of the microenvironmental signals regulating neurogenesis in the adult brain would facilitate the development of such therapeutic approaches. A particularly strong expression of dopamine D(3) receptor mRNA occurs in the proliferative SVZ during prenatal and early postnatal ontogeny. Although its expression diminishes following development, a restricted D(3) receptor expression persists in this region through adulthood, coincident with continued proliferation in this region. Here, we demonstrate a two-fold induction of cell proliferation (BrdU incorporation) in the SVZ and rostral migratory stream of the adult Sprague-Dawley rat brain following intrasubventricular administration of the dopamine D(3) receptor agonist, 7 hydroxy-N,N-di-n-propyl-2-aminotetralin (7-OH-DPAT) for 2 weeks. The number of BrdU-positive cells was elevated ten-fold from very low baseline levels in the neighbouring neostriatum, another region known to express D(3) receptors. These striatal BrdU-positive cells appeared within 3 days following intracerebral infusion of 7-OH-DPAT and were distributed homogeneously throughout the striatum following systemic administration. This suggests that these cells originate from resident progenitor cells rather than the SVZ. Dopamine D(3) receptor activation may serve as a proneuronal differentiation signal as 60-70% of the new cells had neuronal markers following 7-OH-DPAT infusion. These results suggest that the dopamine D(3) receptor may be a good drug target for cell replacement strategies, particularly because of the fact that its expression is almost exclusively limited to the nervous system. PMID- 15128394 TI - Nigral GABAergic inhibition upon mesencephalic dopaminergic cell groups in rats. AB - Synaptic inhibition from the substantia nigra pars reticulata (SNr) to the mesencephalic dopaminergic neurons, which was mediated by gamma (gamma)-amino butyric acid (GABA), was investigated in a midbrain slice preparation of Wistar rats. Whole-cell patch-clamp recordings were used to record synaptic potentials/currents from the dopaminergic neurons (n = 93) located in the retrorubral field (n = 22), the substantia nigra pars compacta (n = 47) and the ventral tegmental area (n = 24). In the presence of ionotropic glutamate receptor antagonists electrical stimulation of the SNr induced inhibitory postsynaptic potentials (IPSPs) and/or currents (IPSCs) in 83 neurons. The IPSPs/IPSCs were comprised early and late components. The early IPSPs/IPSCs were mediated by chloride currents through GABA(A) receptors. The late IPSPs/IPSCs were mediated by potassium currents through GABA(B) receptors. Both GABA(A)- and GABA(B)-IPSPs were amplified by repetitive stimuli with frequencies between 25 and 200 Hz. This frequency range covers the firing frequencies of SNr neurons in vivo. It was observed that an application of a GABA(B) receptor antagonist increased the amplitude of the GABA(A)-IPSPs. The amplification was followed by a rebound depolarization that induced transient firing of dopaminergic neurons. These properties of the IPSPs were common in all of the three dopaminergic nuclei. These results suggest that postsynaptic GABA(A)- and GABA(B)-inhibition contribute to transient and persistent alternations of the excitability of dopaminergic neurons, respectively. These postsynaptic mechanisms may be, in turn, regulated by presynaptic GABA(B)-inhibition. Nigral GABAergic input may provide the temporospatial regulation of the background excitability of mesencephalic dopaminergic systems. PMID- 15128393 TI - Dental pulp cells provide neurotrophic support for dopaminergic neurons and differentiate into neurons in vitro; implications for tissue engineering and repair in the nervous system. AB - Glial cell line-derived neurotrophic factor (GDNF) mRNA is highly expressed by dental pulp cells (DPCs) prior to the initiation of dental pulp innervation. We show that radioactively labelled exogenous GDNF is retrogradely transported from neonatal teeth and vibrissae to the trigeminal neurons, indicating that GDNF acts as a classical neurotrophic factor in the trigeminal system. We also show that DPCs from both rats and humans produce nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and GDNF mRNAs in vitro, promote the survival and phenotypic characteristics of embryonic dopaminergic (DA) neurons and protect DA neurons against the neurotoxin 6-hydroxy-dopamine (6-OHDA) in vitro. By using inhibitory antibodies to NGF, BDNF and GDNF, we show that the promotion of DA neuron survival relates to the production and release of neurotrophic proteins by DPCs in vitro. We suggest that in vivo production of neurotrophic factors by DPCs play roles in tooth innervation. However, continued production of neurotrophic factors by the DPCs might have wider implications. We propose that the dental pulp is a viable source of easily attainable cells with possible potential for development of autologous cell transplantation therapies. We also show that a population of neural crest-derived dental pulp cells acquire clear neuronal morphology and protein expression profile in vitro, indicating the presence of a cell population in the dental pulp with neuronal differentiation capacity that might provide additional benefits when grafted into the CNS. PMID- 15128395 TI - Functional maturation of isolated neural progenitor cells from the adult rat hippocampus. AB - Although neural progenitor cells (NPCs) may provide a source of new neurons to alleviate neural trauma, little is known about their electrical properties as they differentiate. We have previously shown that single NPCs from the adult rat hippocampus can be cloned in the presence of heparan sulphate chains purified from the hippocampus, and that these cells can be pushed into a proliferative phenotype with the mitogen FGF2 [Chipperfield, H., Bedi, K.S., Cool, S.M. & Nurcombe, V. (2002) Int. J. Dev. Biol., 46, 661-670]. In this study, the active and passive electrical properties of both undifferentiated and differentiated adult hippocampal NPCs, from 0 to 12 days in vitro as single-cell preparations, were investigated. Sparsely plated, undifferentiated NPCs had a resting membrane potential of approximately -90 mV and were electrically inexcitable. In > 70%, ATP and benzoylbenzoyl-ATP evoked an inward current and membrane depolarization, whereas acetylcholine, noradrenaline, glutamate and GABA had no detectable effect. In Fura-2-loaded undifferentiated NPCs, ATP and benzoylbenzoyl-ATP evoked a transient increase in the intracellular free Ca(2+) concentration, which was dependent on extracellular Ca(2+) and was inhibited reversibly by pyridoxalphosphate-6-azophenyl-2'-4'-disulphonic acid (PPADS), a P2 receptor antagonist. After differentiation, NPC-derived neurons became electrically excitable, expressing voltage-dependent TTX-sensitive Na(+) channels, low- and high-voltage-activated Ca(2+) channels and delayed-rectifier K(+) channels. Differentiated cells also possessed functional glutamate, GABA, glycine and purinergic (P2X) receptors. Appearance of voltage-dependent and ligand-gated ion channels appears to be an important early step in the differentiation of NPCs. PMID- 15128396 TI - Stage-specific and opposing roles of BDNF, NT-3 and bFGF in differentiation of purified callosal projection neurons toward cellular repair of complex circuitry. AB - Cellular repair of neuronal circuitry affected by neurodegenerative disease or injury may be approached in the adult neocortex via transplantation of neural precursors ("neural stem cells") or via molecular manipulation and recruitment of new neurons from endogenous precursors in situ. A major challenge for potential future approaches to neuronal replacement will be to specifically direct and control progressive differentiation, axonal projection and connectivity of neural precursors along a specific neuronal lineage. This goal will require a progressively more detailed understanding of the molecular controls over morphologic differentiation of specific neuronal lineages, including neurite outgrowth and elongation, in order to accurately permit and direct proper neuronal integration and connectivity. Here, we investigate controls over the morphologic differentiation of a specific prototypical lineage of cortical neurons: callosal projection neurons (CPN). We highly enriched CPN to an essentially pure population, and cultured them at three distinct stages of development from embryonic and postnatal mouse cortex by retrograde fluorescence labelling, followed by fluorescence-activated cell sorting. We find that specific peptide growth factors exert direct stage-specific positive and negative effects over the morphologic differentiation and process outgrowth of CPN. These effects are distinct from the effects of these growth factors on CPN survival [Catapano et al. (2001)J. Neurosci., 21, 8863-8872]. These data may be critical for the future goal of directing lineage-specific neuronal differentiation of transplanted or endogenous precursors/"stem cells" toward cellular repair of complex cortical circuitry. PMID- 15128397 TI - Ca(2+) signalling and gap junction coupling within and between pigment epithelium and neural retina in the developing chick. AB - Development of the neural retina is controlled in part by the adjacent retinal pigment epithelium (RPE). To understand better the mechanisms involved, we investigated calcium signalling and gap junctional coupling within and between the RPE and the neural retina in embryonic day (E) 5 chick. We show that the RPE and the ventricular zone (VZ) of the neural retina display spontaneous Ca(2+) transients. In the RPE, these often spread as waves between neighbouring cells. In the VZ, the frequency of both Ca(2+) transients and waves was lower than in RPE, but increased two-fold in its presence. Ca(2+) signals occasionally crossed the boundary between the RPE and VZ in either direction. In both tissues, the frequency of propagating Ca(2+) waves, but not of individual cell transients, was reduced by gap junction blockers. Use of the gap junction permeant tracer Neurobiotin showed that neural retina cells are coupled into clusters that span the thickness of the retina, and that RPE cells are both coupled together and to clusters of cells in the neural retina. Immunolabelling for Cx43 showed this gap junction protein is present at the junction between the RPE and VZ and thus could potentially mediate the coupling of the two tissues. Immunolabelling for beta tubulin and vimentin showed that clusters of coupled cells in the neural retina comprised mainly progenitor cells. We conclude that gap junctions between progenitor cells, and between these cells and the RPE, may orchestrate retinal proliferation/differentiation, via the propagation of Ca(2+) or other signalling molecules. PMID- 15128398 TI - Energy failure in astrocytes increases the vulnerability of neurons to spreading depression. AB - A neuroprotective role of astrocytes has been hypothesized, but the mechanism is debated and in vivo evidence is limited. To test this hypothesis, a sublethal stressor (spreading depression) and fluorocitrate (FC), a selective inhibitor of the astrocytic Krebs cycle, were used in urethane-anaesthetized adult rats. Neuronal damage was assessed 24 h after treatment with silver stain and immunoreactivity for a 72-kDa heat-shock protein. ATP levels and mitochondrial aconitase activity, a marker indicating exposure to reactive oxygen species, were measured after 4 and 24 h. Spreading depression alone did not affect ATP levels, mitochondrial aconitase activity, or induce neuronal injury in the cortex. Local or intraventricular injection of FC significantly decreased ATP levels and mitochondrial aconitase activity, but did not produce neuronal damage. In animals receiving injections of FC and then spreading depression, there was evidence of significant neuronal stress and damage. Isocitrate, which bypasses the metabolic inhibition produced by FC, prevented all of the changes seen after the combination of FC and spreading depression. One-hour pretreatment with dimethyl sulfoxide (a scavenger of hydroxyl radicals), deferoxamine (an iron chelator) or fructose-1,6-bisphosphate also blocked inactivation of mitochondrial aconitase, ATP depletion and the neuronal damage induced by FC and spreading depression. These experiments demonstrate that inhibition of the metabolism of astrocytes, with a decrease in ATP levels, will increase the susceptibility of neurons to the stress induced by spreading depression. The neuroprotective effects of dimethyl sulfoxide, deferoxamine and fructose-1,6-bisphosphate suggest that oxidative stress contributes to the neurotoxicity in this situation. PMID- 15128399 TI - Modulation of AMPA currents by D2 dopamine receptors in striatal medium-sized spiny neurons: are dendrites necessary? AB - Glutamatergic afferents from the neocortex constitute the major excitatory input to striatal medium-sized spiny neurons (MSNs). Glutamate's actions on MSNs are modulated by dopamine (DA) through D1 and D2 receptor families. Although D1 modulation of glutamate responses has been well-characterized, the contribution of postsynaptic D2 receptors to alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) responses has not been studied extensively. We examined DA modulation of AMPA currents using whole-cell voltage-clamp recordings of MSNs acutely dissociated and in slices. In dissociated cells, the D2 agonist quinpirole (10 micro m) produced small and inconsistent effects on AMPA currents. The magnitude of the current, as well as its modulation by quinpirole, was related to the dendritic elaboration of the dissociated cell. Thus, quinpirole altered AMPA currents only slightly when few initial dendritic segments were present. The amplitude of the current was greater and quinpirole consistently decreased this current in dissociated cells displaying at least three primary dendrites and several secondary and tertiary dendrites. Cyclothiazide, a compound that prevents AMPA receptor desensitization, greatly increased AMPA currents. In the presence of cyclothiazide, quinpirole also consistently reduced AMPA currents. Finally, in slices, AMPA current amplitude was always reduced after application of quinpirole. Sulpiride, a D2 antagonist, prevented attenuation of AMPA currents in both acutely dissociated neurons and neurons in slices. These results provide evidence that AMPA currents are attenuated by DA via activation of postsynaptic D2 receptors. In addition, they indicate that the dendrites and/or the amplitude of the current are important variables for DA modulation of AMPA currents in MSNs. PMID- 15128401 TI - Organization of GABA receptor alpha-subunit clustering in the developing rat neocortex and hippocampus. AB - We compared the expression and co-expression of alpha1, alpha2, alpha3, and alpha5-subunit protein clusters of the gamma-aminobutyric acid (GABA)(A) receptor in the neocortex and hippocampus of rat at postnatal days (PND) 5-10 and 30-40 in order to understand how inhibitory receptors reorganize during brain maturation. The size, intensity, density and pattern of co-localization of fluorescently tagged subunit clusters were determined in deconvolved digital images using a novel 2D cross-correlational analysis. The cross-correlation analysis allowed an unbiased identification of GABA(A) receptor subunit clusters based on staining intensity. Cluster size increased through development; only the alpha2 clusters in dentate gyrus (DG) decreased in size. alpha5-subunit cluster density either increased or decreased with maturation depending on the brain region. For the other subunits, the cluster density remained rather constant, with noted exceptions (increase in alpha2 clusters in cortical layer 5 but a decrease of alpha3 clusters in hilus). The co-localization of alpha1-subunit with the others was unique and not correlated to overall changes in subunit abundance between developmental epoques. So, although alpha2-subunit expression went up in the DG, the clusters became less co-localized with alpha1. In contrast, alpha5-subunit clusters became more co-localized with alpha1 as the alpha5-subunit expression declined in cortex and CA1. The co-localization of alpha3 with alpha1 also became greater in layer 6. In the adult brain not all clustering was associated with synapses, as many alpha-subunit clusters did not co-localize with synaptophysin. Overall, these data indicate that the regulation of GABA(A) receptor clustering is both synaptic and extrasynaptic, presumably reflecting complex cellular trafficking mechanisms. PMID- 15128402 TI - Sex differences in adult suprachiasmatic nucleus neurons emerging late prenatally in rats. AB - The suprachiasmatic nucleus (SCN) is implicated in the control of circadian rhythms of gonadal function. Although several structures surrounding the SCN are sensitive to the effects of gonadal steroids, similar effects in the SCN remain unclear. For example, there are conflicting data on whether the SCN is sexually differentiated. This study attempted to determine sex differences in the number of SCN cells generated during late gestation, and if testosterone mediates these differences. Pregnant female rats were treated with 5-bromo-2'-deoxyuridine (BrdU; 50 mg/kg) on gestational day 18 (E18), the day when aromatase activity peaks in the developing rat fetus. These animals were also given injections of oil or testosterone propionate (10 mg/0.1 mL peanut oil) from E15 until parturition. Litters were allowed to survive until adulthood and were killed on postnatal day 60 (PN60). Following fixation, brain sections containing the SCN from these rats were processed for BrdU immunocytochemistry. A second set of SCN sections was processed for immunocytochemistry detecting BrdU and some of the cell groups prevalent within the SCN. Data showed that female rats have a higher number of cells labeled with BrdU in the SCN, particularly in the medial and caudal SCN. This sex difference was abolished in animals treated with testosterone during late gestation. Double immunocytochemistry revealed that BrdU labeled cells were neurons expressing calbindin-D28K, vasoactive intestinal peptide and, to a lesser degree, vasopressin. Our results unveiled a previously unknown effect of gonadal steroids on the developing SCN, which may contribute to the emergence of gender-specific circadian rhythms. PMID- 15128400 TI - AMPA receptor-mediated presynaptic inhibition at cerebellar GABAergic synapses: a characterization of molecular mechanisms. AB - A major subtype of glutamate receptors, AMPA receptors (AMPARs), are generally thought to mediate excitation at mammalian central synapses via the ionotropic action of ligand-gated channel opening. It has recently emerged, however, that synaptic activation of AMPARs by glutamate released from the climbing fibre input elicits not only postsynaptic excitation but also presynaptic inhibition of GABAergic transmission onto Purkinje cells in the cerebellar cortex. Although presynaptic inhibition is critical for information processing at central synapses, the molecular mechanisms by which AMPARs take part in such actions are not known. This study therefore aimed at further examining the properties of AMPAR-mediated presynaptic inhibition at GABAergic synapses in the rat cerebellum. Our data provide evidence that the climbing fibre-induced inhibition of GABA release from interneurons depends on AMPAR-mediated activation of GTP binding proteins coupled with down-regulation of presynaptic voltage-dependent Ca(2+) channels. A G(i/o)-protein inhibitor, N-ethylmaleimide, selectively abolished the AMPAR-mediated presynaptic inhibition at cerebellar GABAergic synapses but did not affect AMPAR-mediated excitatory actions on Purkinje cells. Furthermore, both G(i/o)-coupled receptor agonists, baclofen and DCG-IV, and the P/Q-type calcium channel blocker omega-agatoxin IVA markedly occluded the AMPAR mediated inhibition of GABAergic transmission. Conversely, AMPAR activation inhibited action potential-triggered Ca(2+) influx into individual axonal boutons of cerebellar GABAergic interneurons. By suppressing the inhibitory inputs to Purkinje cells, the AMPAR-mediated presynaptic inhibition could thus provide a feed-forward mechanism for the information flow from the cerebellar cortex. PMID- 15128403 TI - Subunit-specific P2X-receptor expression defines chemosensory properties of trigeminal neurons. AB - The facial innervation pattern of trigeminal nerve fibres comprises the innervation of the nasal epithelium, where free trigeminal nerve endings contribute to detection and discrimination of chemical stimuli including odourants. The signal transduction mechanisms in sensory nerve endings underlying perception of chemical stimuli remain widely uncovered. Here, we characterized trigeminal ATP-activated P2X receptors in cultured rat trigeminal neurons and investigated their role in chemoperception. We identified a new subpopulation of neurons lacking typical nociceptive characteristics and expressing homomeric P2X(2) receptors. Using a certain group of chemicals known as trigeminal stimuli we found no direct activation of trigeminal neurons, but a modulation of P2X(2) receptor mediated currents. In contrast, P2X(3) receptor mediated currents of nociceptive trigeminal neurons remained unaffected by the tested chemicals. Therefore, we assume a functional role for the newly identified subpopulation in chemodetection of certain trigeminal stimuli. PMID- 15128404 TI - NQO1 activity in the main and the accessory olfactory systems correlates with the zonal topography of projection maps. AB - The mouse olfactory epithelium (OE) is divided into spatial zones, each containing neurons expressing zone-specific subsets of odorant receptor genes. Likewise, the vomeronasal (VN) organ is organized into apical and basal subpopulations of neurons expressing different VN receptor gene families. Axons projecting from the different OE zones and VN subpopulations form synapses within circumscribed regions in the glomerular layer of the olfactory bulb (OB) and accessory olfactory bulb (AOB), respectively. We here show that mature neurons in one defined zone selectively express NADPH:quinone oxidoreductase (NQO1), an enzyme that catalyses reduction of quinones. Immunohistochemistry and in situ hybridization analyses show non-overlapping expression of NQO1 and the Rb8 neural cell adhesion molecule (RNCAM/OCAM) in OE and axon terminals within glomeruli of the OB. In addition, NQO1 immunoreactivity reveals selective, zone-specific axon fasciculation in the olfactory nerve. VN subpopulations do not show complementary patterns of RNCAM and NQO1 immunoreactivity, instead both genes are co-expressed in apical VN neurons that project to the rostral AOB. These results indicate that one division of both the accessory and the main olfactory projection maps are composed of sensory neurons that are specialized to reduce environmental and/or endogenously produced quinones via an NQO1-dependent mechanism. The role of NQO1 in bioactivation of quinoidal drugs also points to a connection between zone specific NQO1 expression and zone-specific toxicity of certain olfactory toxins. PMID- 15128405 TI - Locus coeruleus activation shortens synaptic drive while decreasing spike latency and jitter in sensorimotor cortex. Implications for neuronal integration. AB - Chronic recording of locus coeruleus (LC) neurons in rat and monkey have pointed out that brief, phasic LC discharges, but not sustained activity, are specifically related to salient stimuli and attention. However, the sensory consequences of phasic activation of the noradrenergic system by a brief conditioning stimulation of the LC have not been fully investigated. This study examined the effect of LC activation on synaptic and neuronal responses to a tactile stimulus in the sensorimotor cortex of the anaesthetized rat, by analysing the fine temporal structure of sensory discharges and current source density profiles recorded from the same electrodes. LC stimulation, with minimal EEG effects, consistently reduced the synaptic input in layers IV and V-VI, by decreasing the amplitude and duration of short-latency current sinks, but not the slope of their early rising phase. Simultaneously, most multiple and single unit excitatory responses were shortened by the suppression of their late component after 25-30 ms, whereas robust temporal facilitation of the early discharge was found for spike latency mean and variance, spike timing and synchronization to the stimulus, but leaving the number of spikes unaffected. These two apparently opposite effects on the synaptic drive and neuronal response are reminiscent of the noradrenergic depression of afferent synaptic potentials observed with an increased neuronal excitability in vitro. They are interpreted as a noradrenergic sharpening of thalamocortical processing consistent with a presumed role of synchronous discharges in perception that would depend on activated states, particularly when LC activity is correlated with vigilance or attention. PMID- 15128406 TI - Plasticity of somatostatin and somatostatin sst2A receptors in the rat dentate gyrus during kindling epileptogenesis. AB - Increasing evidence suggests that somatostatin may control neuronal excitability during epileptogenesis. In the hippocampus, sst2A receptors are likely to mediate somatostatin inhibitory actions but little is known about their status in kindled tissues. In the present study, sst2A receptor and somatostatin immunoreactivity were examined by confocal microscopy in the hippocampus during and after kindling acquisition. In control rats, somatostatin-positive axon terminals were mainly found in the stratum lacunosum moleculare of CA1 area and in the outer molecular layer of the dentate gyrus. sst2A receptor immunoreactivity was diffusely distributed in the strata radiatum and oriens of CA1 and in the stratum moleculare of the dentate gyrus. Immunogold electron microscopy revealed that sst2A receptors were predominantly localized postsynaptically, at the plasma membrane of dendritic shafts and spines of principal neurons. During kindling epileptogenesis, qualitative and semiquantitative analysis revealed a progressive decrease of sst2A immunoreactivity in the outer molecular layer, which was spatially associated with an increase in somatostatin immunoreactivity. No obvious changes in sst2A receptor immunoreactivity were observed in other hippocampal subfields. These results suggest that the decrease of sst2A receptor immunoreactivity in the outer molecular layer reflects receptor down-regulation in distal dendrites of granule cells in response to chronic somatostatin release. Because the sst2A receptor appears to mediate anticonvulsant and antiepileptogenic effects of somatostatin, this may represent a pivotal mechanism contributing to epileptogenesis. PMID- 15128407 TI - Endogenous adenosine modulates epileptiform activity in rat hippocampus in a receptor subtype-dependent manner. AB - The purine nucleoside adenosine is released during seizure activity and exerts an anticonvulsant influence through inhibition of glutamate release and hyperpolarization of neurons via adenosine A(1) receptors. However, activation of adenosine A(2A) and A(3) receptors may counteract the inhibitory effects of A(1) receptors. We have therefore examined the extent to which endogenous adenosine released during seizure activity activates the different adenosine receptor subtypes and the implications for seizure activity in the rat hippocampus in vitro. Brief trains of high-frequency stimulation in nominally Mg(2+)-free artificial cerebrospinal fluid evoked epileptiform activity and resulted in a transient depression of the simultaneously recorded CA1 field excitatory postsynaptic potential. In the presence of 8-cyclopentyl-1,3-dimethylxanthine (CPT), an adenosine A(1) receptor antagonist, the occurrence of spontaneous seizure activity was greatly increased as was the duration and intensity of evoked seizures, whilst the postictal depression of basal synaptic transmission was greatly attenuated. Application of ZM 241385, an adenosine A(2A) receptor antagonist, shortened the duration of epileptiform activity, whereas administration of MRS 1191, an adenosine A(3) receptor antagonist, both decreased the duration and intensity of seizures. Combined application of the A(2A) and A(3) receptor antagonists also resulted in a reduction in seizure duration and intensity. However, no evidence was found for a role for protein kinase C in the regulation of seizure activity by endogenous adenosine. Our data confirm the dominant anticonvulsant role that endogenous and tonic adenosine play via the A(1) receptor, and suggest that the additional adenosine receptor subtypes may compromise this anticonvulsant property through promotion of seizure activity. PMID- 15128408 TI - Habenula lesions cause impaired cognitive performance in rats: implications for schizophrenia. AB - Cognitive impairment is a prominent feature of schizophrenia. Currently there is no well-accepted explanation of the aetiology of this disorder, but recent evidence indicates that dysfunction of the habenula may be involved. We therefore examined whether habenula lesions in Sprague-Dawley rats cause behavioural changes resembling those of schizophrenia. Rats received either habenula lesions, a sham operation or a small lesion of the overlying dorsal hippocampus as a check that effects observed were not due to incidental damage to this structure. As there are alterations of social behaviour, sensorimotor gating and cognition in schizophrenia, we examined comparable behaviours. Social interaction time was measured during a 5-min encounter with a novel juvenile conspecific. Prepulse inhibition of an acoustic startle response, as an index of sensorimotor gating, was measured with prepulses of various amplitudes, and spatial cognitive performance was assessed in the Morris water maze task. Histological analysis showed that habenula lesions substantially damaged both medial and lateral habenula bilaterally while largely sparing neighbouring structures. Assay of choline acetyltransferase (ChAT) in the interpeduncular nucleus terminal region of the habenulo-interpeduncular tract, showed marked reduction (by 80%) in habenula-lesioned animals. Habenula-lesioned rats, but not the control group with small dorsal hippocampus lesions, showed marked impairment of Morris maze performance compared to the sham-operated control group. Social interaction time and prepulse inhibition were not significantly altered in either lesion group. The results are consistent with a role of the habenula in cognition, and with the view that pathology of the habenula may contribute to the cognitive impairments of schizophrenia. PMID- 15128409 TI - The Type IV phosphodiesterase inhibitor rolipram interferes with drug-induced conditioned place preference but not immediate early gene induction in mice. AB - Behavioural effects of psychostimulant and opiate drugs are mediated in part by cAMP pathways operating in the nucleus accumbens. Degradation of cAMP occurs through the action of phosphodiesterases, such as the Type IV phosphodiesterases (PDE4s) that are found throughout the brain. To examine the potential role of PDE4 in reward-mediated behaviour, we measured the effects of rolipram, a PDE4 selective inhibitor, on cocaine (18 mg/kg i.p.) and morphine (5 mg/kg s.c.) conditioned place preference in Swiss Webster mice. Rolipram (0, 0.2 or 1.0 mg/kg i.p.) given 30 min prior to drug administration dose-dependently reduced conditioning due to both cocaine and morphine. However, rolipram did not affect place preference induced by food, nor did it prevent the expression of a previously established place preference conditioned by cocaine or morphine. In a second experiment, rolipram administered 30 min prior to a single cocaine injection (50 mg/kg i.p.), did not alter cocaine-induced c-Fos expression in the caudate putamen or nucleus accumbens core. However, rolipram, but not cocaine, induced c-Fos in the nucleus accumbens shell. These results indicate that elevation of cAMP in neurons that express PDE4s may attenuate the rewarding properties of cocaine and morphine, but does not alter the cocaine signalling cascade that induces c-Fos expression. Thus, PDE4-mediated regulation of cAMP levels could underlie the establishment of reward valence to abused drugs. PMID- 15128410 TI - Extracellular excitatory amino acids increase in the paraventricular nucleus of male rats during sexual activity: main role of N-methyl-d-aspartic acid receptors in erectile function. AB - The concentrations of glutamic and aspartic acids were measured in the dialysate obtained with vertical microdialysis probes implanted into the paraventricular nucleus of the hypothalamus of sexually potent male rats during sexual activity. Animals showed noncontact erections when put in the presence of, and copulated with, a receptive (ovarietomized oestrogen- and progesterone-primed) female rat. The concentrations of glutamic and aspartic acids in the paraventricular dialysate increased by 37 and 80%, respectively, above baseline values during exposure to the receptive female rat and by 55 and 127%, respectively, during copulation. No changes in the concentrations of glutamic and aspartic acids were detected in the paraventricular dialysate when sexually potent male rats were exposed to nonreceptive (ovariectomized not oestrogen- and progesterone-primed) female rats or when impotent male rats were used. The injection into the paraventricular nucleus of the excitatory amino acid receptor antagonist dizocilpine (5 micro g), a noncompetitive N-methyl-d-aspartic acid receptor antagonist, reduced noncontact erections and significantly impaired copulatory activity. The alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid receptor antagonist 6-cyano-7-nitro-quinoxaline-2,3-dione (5 micro g) was also able to impair copulatory activity, but to a much lower extent than dizocilpine. In contrast, (+/-)-2-amino-4-phosphono-butanoic acid, a metabotropic receptor antagonist (5 micro g), was found to be ineffective. These results confirm the involvement of the paraventricular nucleus in the control of erectile function and copulatory behaviour and show that excitatory amino acid concentration increases in the paraventricular nucleus when penile erection occurs in physiological contexts. PMID- 15128411 TI - Hippocampal gene expression profiling across eight mouse inbred strains: towards understanding the molecular basis for behaviour. AB - Mouse inbred strains differ in many aspects of their phenotypes, and it is known that gene expression does so too. This gives us an opportunity to isolate the genetic aspect of variation in expression and compare it to other phenotypic variables. We have investigated these issues using an eight-strain expression profile comparison with four replicates per strain on Affymetrix MGU74av2 GeneChips focusing on one well-defined brain tissue (the hippocampus). We identified substantial strain-specific variation in hippocampal gene expression, with more than two hundred genes showing strain differences by a very conservative criterion. Many such genetically driven differences in gene expression are likely to result in functional differences including differences in behaviour. A large panel of inbred strains could be used to identify genes functionally involved in particular phenotypes, similar to genetic correlation. The genetic correlation between expression profiles and function is potentially very powerful, especially given the current large-scale generation of phenotypic data on multiple strains (the Mouse Phenome Project). As an example, the strongest genetic correlation between more than 200 probe sets showing significant differences among our eight inbred strains and a ranking of these strains by aggression phenotype was found for Comt, a gene known to be involved in aggression. PMID- 15128412 TI - Abnormal fronto-parietal coupling of brain rhythms in mild Alzheimer's disease: a multicentric EEG study. AB - Cholinergic deafferentation/recovery in rats mainly impinges on the fronto parietal coupling of brain rhythms [D. P. Holschneider et al. (1999) Exp. Brain Res., 126, 270-280]. Is this reflected by the functional coupling of fronto parietal cortical rhythms at an early stage of Alzheimer's disease (mild AD)? Resting electroencephalographic (EEG) rhythms were studied in 82 patients with mild AD and in control subjects, such as 41 normal elderly (Nold) subjects and 25 patients with vascular dementia (VaD). Patients with AD and VaD had similar mini mental state evaluation scores of 17-24. The functional coupling was estimated by means of the synchronization likelihood (SL) of the EEG data at electrode pairs, accounting for linear and non-linear components of that coupling. Cortical rhythms of interest were delta (2-4 Hz), theta (4-8 Hz), alpha (1 8-10.5 Hz), alpha 2 (10.5-13 Hz), beta 1 (13-20 Hz), beta 2 (20-30 Hz) and gamma (30-40 Hz). A preliminary data analysis (Nold) showed that surface Laplacian transformation of the EEG data reduced the values of SL, possibly because of the reduction of influences due to head volume conduction. Therefore, the final analysis was performed on Laplacian-transformed EEG data. The SL was dominant at alpha 1 band in all groups. Compared with the Nold subjects, patients with VaD and mild AD presented a marked reduction of SL at both fronto-parietal (delta-alpha) and inter-hemispherical (delta-beta) electrode pairs. The feature distinguishing the patients with mild AD with respect to patients with VaD groups was a more prominent reduction of fronto-parietal alpha 1 SL. These results suggest that mild AD is characterized by an abnormal fronto-parietal coupling of the dominant human cortical rhythm at 8-10.5 Hz. PMID- 15128413 TI - Dissociating brain regions controlling the temporal and ordinal structure of learned movement sequences. AB - We used functional magnetic resonance imaging to investigate if different brain regions are controlling the temporal and ordinal structure of movement sequences during performance. Human subjects performed overlearned spatiotemporal sequences of key-presses using the right index finger. Under different conditions, the temporal and the ordinal structure of the sequences were varied systematically in relation to each other, using a factorial design: COMBINED had a rhythm of eight temporal intervals and a serial order of eight keys; TEMPORAL had an eight interval rhythm produced on one key; ORDINAL had an isochronous rhythm and an eight-key serial order; two control conditions had an isochronous pulse performed on one or two keys, respectively. Brain regions involved in rhythmic and ordinal control of the sequences were revealed by analysing main effect contrasts for the corresponding factors. TEMPORAL and ORDINAL were also compared directly to test for significant differences. A dissociation was found between largely the presupplementary motor area, the right inferior frontal gyrus and precentral sulcus, and the bilateral superior temporal gyri, involved in temporal control, and lateral fronto-parietal areas, the basal ganglia and the cerebellum, which were implicated in ordinal control. The vermis and the superior colliculus were the only regions with an activity increase specifically related to combining long temporal and ordinal sequences. We conclude that humans use different brain networks for temporal and ordinal sequence control, and that the performance of combined sequences activates both networks, the medial cerebellum, and the superior colliculus. PMID- 15128414 TI - Hearing syllables by seeing visual stimuli. AB - Currently it is discussed whether the same cortical areas are activated during the imagination of as during the actual presentation of specific stimuli. Some argue that mostly the secondary but not the primary sensory areas are active during imagination. Using functional magnetic resonance imaging we explored whether auditory verbal imagery of syllables has sufficient power to evoke haemodynamic responses in the auditory cortex. To overcome the detrimental effects of scanner noise, one group of subjects was trained to vividly imagine hearing a syllable while a flashlight was presented. A control group did not receive this training. We found that only the trained group revealed haemodynamic responses in the auditory cortex during auditory imagination while the control group showed no activation within the auditory cortex. Peak activations during auditory verbal imagery are located bilaterally within the superior temporal gyrus region in the vicinity of the planum temporale. While these secondary auditory areas are active during auditory verbal imagery, there was no activation in Heschl's gyrus. We hypothesize that auditory verbal imagery is associated with haemodynamic responses in secondary auditory and not primary auditory areas. PMID- 15128415 TI - Left hemisphere motor facilitation in response to manual action sounds. AB - Previous studies indicate that the motor areas of both hemispheres are active when observing actions. Here we explored how the motor areas of each hemisphere respond to the sounds associated with actions. We used transcranial magnetic stimulation (TMS) to measure motor corticospinal excitability of hand muscles while listening to sounds. Sounds associated with bimanual actions produced greater motor corticospinal excitability than sounds associated with leg movements or control sounds. This facilitation was exclusively lateralized to the left hemisphere, the dominant hemisphere for language. These results are consistent with the hypothesis that action coding may be a precursor of language. PMID- 15128416 TI - Quality of life of patients with chronic lymphocytic leukemia: results of a longitudinal investigation over 1 yr. AB - OBJECTIVES: The aim of this longitudinal study was to determine the long-term quality of life (QoL) of patients with chronic lymphocytic leukemia (CLL) and to investigate the relationship between QoL and sociodemographic and clinical parameters. METHODS: Ninety-seven patients suffering from CLL were asked to complete the European Organization for Research and Treatment of Cancer Quality of Life Questionnaire (EORTC QLQ-C30) four times over a period of 1 yr. Clinical data on disease and treatment characteristics were collected from medical records. For the purpose of comparison, EORTC QLQ-C30 scores were collected from 152 age- and gender-matched healthy controls. RESULTS: Seventy-six patients (age: median 68 yr, range 41-89) returned one or more questionnaires and were included in the statistical analysis. Compared with healthy controls CLL patients reported a lower QoL in almost all domains. No differences regarding QoL could be observed between CLL patients who had already received chemotherapy and those who had not. Moreover, female CLL patients were found to have remarkably lower QoL scores in the areas of emotional and social functioning than male patients. CONCLUSION: Patients suffering from CLL could have their QoL improved by more effective symptom management and psycho-oncological support. This could focus on specific symptoms such as fatigue and might have particular benefits for female patients with respect to their emotional and social well-being. PMID- 15128417 TI - A narrow deletion of 7q is common to HCL, and SMZL, but not CLL. AB - To further characterise the genetic background of the two closely related B lymphocytic malignancies hairy cell leukaemia (HCL), and splenic marginal zone lymphoma (SMZL) we have identified characteristic copy number imbalances by comparative genomic hybridisation (CGH). Based on these findings, areas of special interest were fine mapped, and relevant probes constructed for use in interphase-fluorescence in situ hybridisation (FISH) investigations. Thus, using the CGH data from 52 HCL and 61 SMZL patients, we identified the characteristic profiles of copy number imbalances for both diseases. These were a gain of 5q13 31 (19%) and loss of 7q22-q35 (6%) for HCL, and gain of 3q25 (28%), loss of 7q31 (16%), and gain of 12q15 (16%) for SMZL. A partial loss of 7q unusual for low malignant B-cell diseases was found to be common to the two diseases. This loss was therefore fine mapped with BAC/PAC clones. Fine mapping revealed that in SMZL the minimal lost region covers 11.4 Mb spanning from 7q31.33 to 7q33 located between sequence tagged site (STS)-markers SHGC-3275 and D7S725. This area was distinct from the commonly deleted 7q region of myelodysplastic syndrome/acute myeloid leukaemia (MDS/AML). A FISH probe specific for the 7q region was constructed. Using this probe in an interphase-FISH investigation we showed the minimal lost 7q-region of HCL and SMZL to be one and the same. In one HCL case, this investigation furthermore showed the extent of the deleted region to be below the detection limit of CGH, whereas interphase-FISH screening of 36 chronic lymphocytic leukaemia (CLL) cases showed no deletion of the 7q area. In conclusion, we have identified characteristic profiles of copy number imbalances in HCL and SMZL and fine mapped the minimal extent of a commonly lost 7q area of special interest. We hypothesise that this region may contain (a) gene(s) important for the pathology of HCL and SMZL. PMID- 15128418 TI - Common and rare side-effects of low-dose thalidomide in multiple myeloma: focus on the dose-minimizing peripheral neuropathy. AB - OBJECTIVES: Thalidomide has demonstrated a remarkable efficacy in the treatment of multiple myeloma but its use may cause several toxicities. We have investigated the common and rare side-effects, especially analysing peripheral neuropathy, in order to optimise the thalidomide dose for minimizing this harmful side-effect. METHODS: Fifty-nine patients were treated with thalidomide alone or combined with oral melphalan. The median age was 69 yr. The initial dose of thalidomide was 100 mg/day increasing weekly by 100 mg increments until a maximum dose of 400 mg was attained. Melphalan was administered at a dose of 0.20 mg/kg/d for 4 d every 28 d. RESULTS AND CONCLUSIONS: Nearly one-fourth of patients discontinued thalidomide because of toxicity. Constipation (71%), somnolence (36%) and fatigue (20%) were the most common side-effects and they were not dose dependent. Peripheral neuropathy occurred in 39% of patients and a thalidomide median daily dose of more than 150 mg was significantly associated with higher frequency and actuarial risk of peripheral neuropathy without improving the response rate. Deep venous thrombosis was observed in 7% of patients and other side-effects were rare. In patients with advanced multiple myeloma we found that a thalidomide daily dose of 150 mg minimizes peripheral neuropathy without jeopardizing response and survival. PMID- 15128419 TI - Angiogenesis in chronic myeloproliferative diseases detected by CD34 expression. AB - Increased bone marrow angiogenesis estimated as bone marrow microvessel density (MVD), or as serum angiogenic factor levels and/or immunohistochemical expression of these factors in bone marrow biopsy has been demonstrated in a variety of hematological disorders including chronic myeloproliferative diseases (MPDs). The aim of this study was to investigate the MVD in 25 cases of myelofibrosis with myeloid metaplasia (MMM). MVD was estimated by CD34 immunohistochemical expression in bone marrow biopsies. A control group of 27 patients without bone marrow disease, eight cases of polycythemia vera (PV), 41 cases of essential thrombocythemia (ET) and nine cases of chronic myeloid leukemia (CML) were also studied. Moreover, in cases with MMM, MVD was correlated with clinical, laboratory, histological parameters and the outcome of the patients. Our study confirmed a significantly higher degree of angiogenesis in MMM, PV, ET and CML compared with controls (P < 0.001, P = 0.0007, P < 0.001 and P = 0.0008, respectively). Angiogenesis was higher in MMM than PV, ET and CML cases (P < 0.001, P < 0.001 and P = 0.008). Increased angiogenesis was correlated with hypercatabolic symptoms in MMM patients (P = 0.009). No correlation with other clinicopathological parameters or clinical outcome was found. However, definitive conclusions regarding the prognostic value of increased angiogenesis may require additional follow-up and a larger group of patients. PMID- 15128420 TI - Levels of soluble angiogenin in chronic myeloid malignancies: clinical implications. AB - Angiogenesis is critical for the clinical progression of haematopoietic malignancies and depends on angiogenic factors. Angiogenin is a powerful factor produced by neoplastic cells and host microenvironment. High levels of soluble angiogenin (sAng) correlate with a poor prognosis in patients affected by acute myeloid leukaemia and myelodysplastic syndromes, but no data are available on sAng in chronic myeloproliferative disorders (CMD). Therefore, in this study we investigated the clinical significance of the angiogenin in sera of patients with chronic myeloid leukaemia (CML) (n = 14) or essential thrombocythaemia (ET) (n = 20), and correlated them with those of soluble transforming growth factor-beta(1) (sTGF beta(1)). Enzyme-linked immunosorbent assay detected (P < 0.05) higher levels of sAng in CMD compared with healthy subjects (1026.74 +/- 464.60 pg/mL and 196.00 +/- 39.90 pg/mL, respectively). The highest levels of sAng were detected in CML patients (1349.23 +/- 549.55 pg/mL). Interestingly, CML patients who achieved haematological remission after interferon therapy showed circulating levels of angiogenin significantly (P < 0.05) decreased when compared with those at diagnosis. In ET patients, levels of angiogenin (889.34 +/- 267.66 pg/mL) and sTGF beta(1) (76.69 +/-6.08 pg/mL) were higher (P < 0.05) compared with healthy controls (57.93 +/- 19.39 pg/mL). No correlation was found between levels of sAng and levels of sTGF beta(1) or platelet count among ET patients. Our results show for the first time that elevated blood levels of angiogenin feature chronic myeloid malignancies, suggesting a role of angiogenin in the pathogenesis of these diseases. PMID- 15128421 TI - The role of the STAT5 proteins in the proliferation and apoptosis of the CML and AML cells. AB - OBJECTIVES: The STAT5 proteins are activated by many haematological cytokines and growth factors. They regulate cell cycle, apoptosis and proliferation of different cells via the influence on gene transcription. Because STAT5s are constitutively activated in certain haematooncologic diseases, they are suggested to play an important role in leukaemogenesis. However, the real function of these proteins in haematopoietic cell transformation and proliferation is not clear enough. The aim of this study was to evaluate the influence of suppression of STAT5A and STAT5B expression on the clonogenicity and apoptosis of the chronic myeloid leukaemia (CML) and acute myeloid leukaemia (AML) cells. MATERIAL AND METHODS: Blast cells from 34 newly diagnosed patients with CML and AML were used in our experiments. Antisense oligodeoxynucleotides (ODNs) were applied to block STAT5A and STAT5B at the mRNA level and the RT-PCR method was used to study STAT5 mRNA expression in the cells after incubation with ODNs. Moreover, Western blot analysis of the STAT5 proteins was performed. The effect of ODN pretreatment on cell clonogenicity in methylocellulose cultures was examined according to the type of oligodeoxynucleotide and the time of exposure. The induction of apoptosis in cells was also estimated by the Annexin V/PI staining and the TUNEL method using flow cytometry. RESULTS: Perturbation of STAT5 expression decreased proliferative potential of the CML and the AML blasts as well as enhanced their apoptosis (P < 0.05). CONCLUSIONS: Our studies showed that the STAT5 proteins may be critical in the regulation of growth and apoptosis of the CML and AML leukaemic cells. PMID- 15128422 TI - Outcome of allogeneic peripheral blood stem cell transplantation using matched sibling donors in patients with high-risk hematological diseases. AB - Although the use of peripheral blood stem cells instead of bone marrow is still a matter of debate in transplantation from HLA-identical sibling donors, allogeneic peripheral blood stem cell transplantation (PBSCT), with a stronger graft-versus leukemia (GVL) effect, may be preferable as a source of stem cells, especially in the case of advanced hematologic diseases. As such, the current paper reports on the outcomes of 27 consecutive patients with high-risk hematologic diseases treated with allogeneic PBSCT. The median dose of CD34+, CD3+ cells, and MNC infused was 8.18 x 10(6)/kg (range: 2.78-14.93), 1.50 x 10(8)/kg (range: 0.06 4.25), and 7.17 x 10(8)/kg (range: 0.95-15.85), respectively. The median time taken for the ANC and platelets to reach 500 and 20,000 x 10(6)/microL was 15 (range: 9-25) and 16 d (range: 10-56), respectively. Three patients (11.1%) experienced transplant-related mortality within 90 d of transplantation, and 15 (62.5%) of 24 evaluated patients developed chronic graft-versus-host disease (GVHD; six limited, nine extensive). There was a significant difference in overall survival (OS) between the group with chronic GVHD and the group without chronic GVHD (P = 0.0253). The causes of death included relapse (six cases) and non-relapse mortality (infection: four cases, chronic GVHD-related death: three cases). The 4-yr OS rate and disease-free survival rate was 43.3 +/- 10.9% and 35.8 +/- 10.2%, respectively. Accordingly, chronic GVHD was found to have a positive role in patients with high-risk hematologic diseases that received allogeneic PBSCT. PMID- 15128423 TI - The C-->G transition in the alpha 2-globin gene of a normal alpha alpha chromosome is responsible for the Hb G-Philadelphia variant in Sardinians. AB - Sequencing of alpha-globin genes of 18 Sardinian heterozygotes for the Hb G Philadelphia [alpha 68(E17)Asn-->Lys] variant, with four active alpha genes and circulating level of the variant of about 27%, showed the AAC-->AAG change at codon 68 of the alpha 2-globin gene (alpha(G)alpha/alpha alpha). Two heterozygotes with level of about 37% were the carriers of the same mutation on the same alpha 2 gene, and of the alpha 2 alpha 1 hybrid gene, because of the 3.7 kb deletion, in trans (alpha(G)alpha/-alpha(3.7)). In Black people, the same C- >G mutation occurs on the hybrid gene (-alpha(G)3.7), whereas in Caucasians the Lys for Asn change is because of the C-->A transversion occurring on the alpha 2 gene of a normal alpha alpha arrangement. The identification of the C-->G mutation on the normal alpha alpha chromosome points to an undescribed genotype for this rather common variant, which is probably because of the high rate of recombination between the duplicated alpha-globin genes. PMID- 15128424 TI - Sustained cytogenetic response after discontinuation of imatinib mesylate in a patient with chronic myeloid leukaemia. AB - A 58-yr-old woman was diagnosed with Ph(+) chronic myeloid leukaemia in May 2001. She was initially treated with hydroxyurea and subsequently with interferon-alpha (IFN-alpha). Imatinib mesylate was started in April 2002 after failure of IFN alpha to induce a cytogenetic response. The patient remained on treatment with imatinib mesylate for 3 months during which she suffered daily fever resulting in discontinuation of the treatment. Response evaluation performed shortly after discontinuing imatinib mesylate revealed a complete cytogenetic remission and a substantial molecular response. Fifteen months later, she was still enjoying a major cytogenetic response. This case illustrates that a short course of imatinib mesylate may result in a sustained haematological and cytogenetic response. PMID- 15128425 TI - Fatal Evans' syndrome after matched unrelated donor transplantation for hyper-IgM syndrome. AB - A 3 and 1/2-yr-old boy underwent matched unrelated stem cell transplantation (SCT) for hyper-IgM syndrome. He developed acute and chronic skin graft-vs.-host disease (GVHD). Ten months following SCT he presented with severe hemolytic anemia and thrombocytopenia (Evans' syndrome). Treatment included high-dose steroids, intravenous immunoglobulins, cyclosporine, mycophenolate mofetil, chemotherapeutic agents (cyclophosphamide, vincristine, VP-16), immunoadsorption, and anti-CD20 and anti-CD52 monoclonal antibodies without response. The patient died 16 months after SCT. PMID- 15128426 TI - Reduced intensity allogeneic stem cell transplantation for systemic primary amyloidosis refractory to high-dose melphalan. AB - Complete elimination of the plasma cell dyscrasia is a rational therapeutic goal, as intercepting supply of precursor protein is a necessary condition for a major regression of amyloid deposits. High-dose melphalan with autologous stem cell transplantation has shown the ability to induce complete hematological response (HR) along with recovery of organ dysfunction. However, the rate of HR with this treatment rarely exceeds 40%. We describe here the first known case of successful reduced intensity allogeneic stem cell transplantation (RIST) for a patient with primary amyloidosis complicated with nephrotic syndrome but without cardiac disease, who had obtained only partial HR by high-dose melphalan with autologous stem cell transplantation. RIST may be feasible and be capable of achieving complete HR along with recovery from nephrotic syndrome with acceptable toxicity. PMID- 15128427 TI - Acquired pure megakaryocytic aplasia: a separate haematological disease entity or a syndrome with multiple causes? AB - We report the case of a patient with acquired pure megakaryocytic aplasia. Until today, less than 20 cases of acquired pure megakaryocytic aplasia have been reported and the disease aetiology still seems to be unclear. This report summarizes the published data concerning possible aetiologies, treatment options and outcome of patients with acquired pure megakaryocytic aplasia. Furthermore, this case report presents an example for a possible disease progression. PMID- 15128428 TI - Effective treatment of life-threatening bleeding with recombinant activated factor VII in a patient with acute promyelocytic leukaemia. PMID- 15128429 TI - Simultaneous quantitative and allele-specific expression analysis with real competitive PCR. AB - BACKGROUND: For a diploid organism such as human, the two alleles of a particular gene can be expressed at different levels due to X chromosome inactivation, gene imprinting, different local promoter activity, or mRNA stability. Recently, imbalanced allelic expression was found to be common in human and can follow Mendelian inheritance. Here we present a method that employs real competitive PCR for allele-specific expression analysis. RESULTS: A transcribed mutation such as a single nucleotide polymorphism (SNP) is used as the marker for allele-specific expression analysis. A synthetic mutation created in the competitor is close to a natural mutation site in the cDNA sequence. PCR is used to amplify the two cDNA sequences from the two alleles and the competitor. A base extension reaction with a mixture of ddNTPs/dNTP is used to generate three oligonucleotides for the two cDNAs and the competitor. The three products are identified and their ratios are calculated based on their peak areas in the MALDI-TOF mass spectrum. Several examples are given to illustrate how allele-specific gene expression can be applied in different biological studies. CONCLUSIONS: This technique can quantify the absolute expression level of each individual allele of a gene with high precision and throughput. PMID- 15128430 TI - TMF is a golgin that binds Rab6 and influences Golgi morphology. AB - BACKGROUND: Golgins are coiled-coil proteins associated with the Golgi apparatus, that are believed to be involved in the tethering of vesicles and the stacking of cisternae, as well as other functions such as cytoskeletal association. Many are peripheral membrane proteins recruited by GTPases. Several have been described in animal cells, and some in yeast, but the relationships between golgins from different species can be hard to define because although they share structural features, their sequences are not well conserved. RESULTS: We show here that the yeast protein Sgm1, previously shown to be recruited to the Golgi by the GTPase Ypt6, binds to Ypt6:GTP via a conserved 100-residue coiled-coil motif that can be identified in a wide range of eukaryotes. The mammalian equivalent of Sgm1 is TMF/ARA160, a protein previously identified in various screens as a putative transcription or chromatin remodelling factor. We show that it is a Golgi protein, and that it binds to the three known isoforms of the Ypt6 homologue Rab6. Depletion of the protein by RNA interference in rat NRK cells results in a modest dispersal of Golgi membranes around the cell, suggesting a role for TMF in the movement or adherence of Golgi stacks. CONCLUSION: We have identified TMF as an evolutionarily conserved golgin that binds Rab6 and contributes to Golgi organisation in animal cells. PMID- 15128431 TI - Resolution of large and small differences in gene expression using models for the Bayesian analysis of gene expression levels and spotted DNA microarrays. AB - BACKGROUND: The detection of small yet statistically significant differences in gene expression in spotted DNA microarray studies is an ongoing challenge. Meeting this challenge requires careful examination of the performance of a range of statistical models, as well as empirical examination of the effect of replication on the power to resolve these differences. RESULTS: New models are derived and software is developed for the analysis of microarray ratio data. These models incorporate multiplicative small error terms, and error standard deviations that are proportional to expression level. The fastest and most powerful method incorporates additive small error terms and error standard deviations proportional to expression level. Data from four studies are profiled for the degree to which they reveal statistically significant differences in gene expression. The gene expression level at which there is an empirical 50% probability of a significant call is presented as a summary statistic for the power to detect small differences in gene expression. CONCLUSIONS: Understanding the resolution of difference in gene expression that is detectable as significant is a vital component of experimental design and evaluation. These small differences in gene expression level are readily detected with a Bayesian analysis of gene expression level that has additive error terms and constrains samples to have a common error coefficient of variation. The power to detect small differences in a study may then be determined by logistic regression. PMID- 15128432 TI - Statistical monitoring of weak spots for improvement of normalization and ratio estimates in microarrays. AB - BACKGROUND: Several aspects of microarray data analysis are dependent on identification of genes expressed at or near the limits of detection. For example, regression-based normalization methods rely on the premise that most genes in compared samples are expressed at similar levels and therefore require accurate identification of nonexpressed genes (additive noise) so that they can be excluded from the normalization procedure. Moreover, key regulatory genes can maintain stringent control of a given response at low expression levels. If arbitrary cutoffs are used for distinguishing expressed from nonexpressed genes, some of these key regulatory genes may be unnecessarily excluded from the analysis. Unfortunately, no accurate method for differentiating additive noise from genes expressed at low levels is currently available. RESULTS: We developed a multistep procedure for analysis of mRNA expression data that robustly identifies the additive noise in a microarray experiment. This analysis is predicated on the fact that additive noise signals can be accurately identified by both distribution and statistical analysis. CONCLUSIONS: Identification of additive noise in this manner allows exclusion of noncorrelated weak signals from regression-based normalization of compared profiles thus maximizing the accuracy of these methods. Moreover, genes expressed at very low levels can be clearly identified due to the fact that their expression distribution is stable and distinguishable from the random pattern of additive noise. PMID- 15128433 TI - GeneOrder3.0: software for comparing the order of genes in pairs of small bacterial genomes. AB - BACKGROUND: An increasing number of whole viral and bacterial genomes are being sequenced and deposited in public databases. In parallel to the mounting interest in whole genomes, the number of whole genome analyses software tools is also increasing. GeneOrder was originally developed to provide an analysis of genes between two genomes, allowing visualization of gene order and synteny comparisons of any small genomes. It was originally developed for comparing virus, mitochondrion and chloroplast genomes. This is now extended to small bacterial genomes of sizes less than 2 Mb. RESULTS: GeneOrder3.0 has been developed and validated successfully on several small bacterial genomes (ca. 580 kb to 1.83 Mb) archived in the NCBI GenBank database. It is an updated web-based "on-the-fly" computational tool allowing gene order and synteny comparisons of any two small bacterial genomes. Analyses of several bacterial genomes show that a large amount of gene and genome re-arrangement occurs, as seen with earlier DNA software tools. This can be displayed at the protein level using GeneOrder3.0. Whole genome alignments of genes are presented in both a table and a dot plot. This allows the detection of evolutionary more distant relationships since protein sequences are more conserved than DNA sequences. CONCLUSIONS: GeneOrder3.0 allows researchers to perform comparative analysis of gene order and synteny in genomes of sizes up to 2 Mb "on-the-fly." AVAILABILITY: http://binf.gmu.edu/genometools.html and http://pasteur.atcc.org:8050/GeneOrder3.0. PMID- 15128434 TI - Integrated allosteric regulation in the S. cerevisiae carbamylphosphate synthetase - aspartate transcarbamylase multifunctional protein. AB - BACKGROUND: The S. cerevisiae carbamylphosphate synthetase - aspartate transcarbamylase multifunctional protein catalyses the first two reactions of the pyrimidine pathway. In this organism, these two reactions are feedback inhibited by the end product UTP. In the present work, the mechanisms of these integrated inhibitions were studied. RESULTS: The results obtained show that the inhibition is competitive in the case of carbamylphosphate synthetase and non-competitive in the case of aspartate transcarbamylase. They also identify the substrate whose binding is altered by this nucleotide and the step of the carbamylphosphate synthetase reaction which is inhibited. Furthermore, the structure of the domains catalyzing these two reactions were modelled in order to localize the mutations which, specifically, alter the aspartate transcarbamylase sensitivity to the feedback inhibitor UTP. Taken together, the results make it possible to propose a model for the integrated regulation of the two activities of the complex. UTP binds to a regulatory site located in the vicinity of the carbamylphosphate synthetase catalytic subsite which catalyzes the third step of this enzyme reaction. Through a local conformational change, this binding decreases, competitively, the affinity of this site for the substrate ATP. At the same time, through a long distance signal transmission process it allosterically decreases the affinity of the aspartate transcarbamylase catalytic site for the substrate aspartate. CONCLUSION: This investigation provides informations about the mechanisms of allosteric inhibition of the two activities of the CPSase-ATCase complex. Although many allosteric monofunctional enzymes were studied, this is the first report on integrated allosteric regulation in a multifunctional protein. The positions of the point mutations which specifically abolish the sensitivity of aspartate transcarbamylase to UTP define an interface between the carbamylphosphate synthetase and aspartate transcarbamylase domains, through which the allosteric signal for the regulation of aspartate transcarbamylase must be propagated. PMID- 15128435 TI - The ascent of man? AB - Anyone who cares about the moral and social implications of genomics, genetic engineering and biotechnology should read Michael J. Sandel's article, 'The Case Against Perfection', in the April 2004 issue of The Atlantic Monthly. PMID- 15128436 TI - Progress towards mapping the universe of protein folds. AB - Although the precise aims differ between the various international structural genomics initiatives currently aiming to illuminate the universe of protein folds, many selectively target protein families for which the fold is unknown. How well can the current set of known protein families and folds be used to estimate the total number of folds in nature, and will structural genomics initiatives yield representatives for all the major protein families within a reasonable time scale? PMID- 15128437 TI - Insights from the rat genome sequence. AB - The availability of the rat genome sequence, and detailed three-way comparison of the rat, mouse and human genomes, is revealing a great deal about mammalian genome evolution. Together with recent developments in cloning technologies, this heralds an important phase in rat research. PMID- 15128438 TI - The mechanism of prion strain propagation. AB - Studies of mammalian prion diseases such as bovine spongiform encephalopathy have suggested that different strains consist of prion proteins with different conformations. Two recent studies of yeast prions have now formally demonstrated that multiple stable protein conformations are the basis of strain variation. PMID- 15128439 TI - Maize DNA-sequencing strategies and genome organization. AB - A large amount of repetitive DNA complicates the assembly of the maize genome sequence. Genome-filtration techniques, such as methylation-filtration and high CoT separation, enrich gene sequences in genomic libraries. These methods may provide a low-cost alternative to whole-genome sequencing for maize and other complex genomes. PMID- 15128440 TI - Sirtuins: Sir2-related NAD-dependent protein deacetylases. AB - Silent information regulator 2 (Sir2) proteins, or sirtuins, are protein deacetylases dependent on nicotine adenine dinucleotide (NAD) and are found in organisms ranging from bacteria to humans. In eukaryotes, sirtuins regulate transcriptional repression, recombination, the cell-division cycle, microtubule organization, and cellular responses to DNA-damaging agents. Sirtuins have also been implicated in regulating the molecular mechanisms of aging. The Sir2 catalytic domain, which is shared among all sirtuins, consists of two distinct domains that bind NAD and the acetyl-lysine substrate, respectively. In addition to the catalytic domain, eukaryotic sirtuins contain variable amino- and carboxy terminal extensions that regulate their subcellular localizations and catalytic activity. PMID- 15128441 TI - A European focus on proteomics. PMID- 15128442 TI - Discovering the seeds of diversity in plant genomes. PMID- 15128443 TI - The changing face of genomics. PMID- 15128444 TI - A novel family of P-loop NTPases with an unusual phyletic distribution and transmembrane segments inserted within the NTPase domain. AB - BACKGROUND: Recent sequence-structure studies on P-loop-fold NTPases have substantially advanced the existing understanding of their evolution and functional diversity. These studies provide a framework for characterization of novel lineages within this fold and prediction of their functional properties. RESULTS: Using sequence profile searches and homology-based structure prediction, we have identified a previously uncharacterized family of P-loop NTPases, which includes the neuronal membrane protein and receptor tyrosine kinase substrate Kidins220/ARMS, which is conserved in animals, the F-plasmid PifA protein involved in phage T7 exclusion, and several uncharacterized bacterial proteins. We refer to these (predicted) NTPases as the KAP family, after Kidins220/ARMS and PifA. The KAP family NTPases are sporadically distributed across a wide phylogenetic range in bacteria but among the eukaryotes are represented only in animals. Many of the prokaryotic KAP NTPases are encoded in plasmids and tend to undergo disruption to form pseudogenes. A unique feature of all eukaryotic and certain bacterial KAP NTPases is the presence of two or four transmembrane helices inserted into the P-loop NTPase domain. These transmembrane helices anchor KAP NTPases in the membrane such that the P-loop domain is located on the intracellular side. We show that the KAP family belongs to the same major division of the P-loop NTPase fold with the AAA+, ABC, RecA-like, VirD4-like, PilT-like, and AP/NACHT-like NTPase classes. In addition to the KAP family, we identified another small family of predicted bacterial NTPases, with two transmembrane helices inserted into the P-loop domain. This family is not specifically related to the KAP NTPases, suggesting independent acquisition of the transmembrane helices. CONCLUSIONS: We predict that KAP family NTPases function principally in the NTP-dependent dynamics of protein complexes, especially those associated with the intracellular surface of cell membranes. Animal KAP NTPases, including Kidins220/ARMS, are likely to function as NTP dependent regulators of the assembly of membrane-associated signaling complexes involved in neurite growth and development. One possible function of the prokaryotic KAP NTPases might be in the exclusion of selfish replicons, such as viruses, from the host cells. Phylogenetic analysis and phyletic patterns suggest that the common ancestor of the animals acquired a KAP NTPase via lateral transfer from bacteria. However, an earlier transfer into eukaryotes followed by multiple losses in several eukaryotic lineages cannot be ruled out. PMID- 15128446 TI - Detection of evolutionarily stable fragments of cellular pathways by hierarchical clustering of phyletic patterns. AB - BACKGROUND: Phyletic patterns denote the presence and absence of orthologous genes in completely sequenced genomes and are used to infer functional links between genes, on the assumption that genes involved in the same pathway or functional system are co-inherited by the same set of genomes. However, this basic premise has not been quantitatively tested, and the limits of applicability of the phyletic-pattern method remain unknown. RESULTS: We characterized a hierarchy of 3,688 phyletic patterns encompassing more than 5,000 known protein coding genes from 66 complete microbial genomes, using different distances, clustering algorithms, and measures of cluster quality. The most sensitive set of parameters recovered 223 clusters, each consisting of genes that belong to the same metabolic pathway or functional system. Fifty-six clusters included unexpected genes with plausible functional links to the rest of the cluster. Only a small percentage of known pathways and multiprotein complexes are co-inherited as one cluster; most are split into many clusters, indicating that gene loss and displacement has occurred in the evolution of most pathways. CONCLUSIONS: Phyletic patterns of functionally linked genes are perturbed by differential gains, losses and displacements of orthologous genes in different species, reflecting the high plasticity of microbial genomes. Groups of genes that are co inherited can, however, be recovered by hierarchical clustering, and may represent elementary functional modules of cellular metabolism. The phyletic patterns approach alone can confidently predict the functional linkages for about 24% of the entire data set. PMID- 15128445 TI - Multi-tissue gene-expression analysis in a mouse model of thyroid hormone resistance. AB - BACKGROUND: Resistance to thyroid hormone (RTH) is caused by mutations of the thyroid hormone receptor beta (TRbeta) gene. To understand the transcriptional program underlying TRbeta mutant-induced phenotypic expression of RTH, cDNA microarrays were used to profile the expression of 11,500 genes in a mouse model of human RTH. RESULTS: We analyzed transcript levels in cerebellum, heart and white adipose tissue from a knock-in mouse (TRbetaPV/PV mouse) that harbors a human mutation (referred to as PV) and faithfully reproduces human RTH. Because TRbetaPV/PV mice have elevated thyroid hormone (T3), to define T3-responsive genes in the context of normal TRbeta, we also analyzed T3 effects in hyperthyroid wild-type gender-matched littermates. Microarray analysis revealed 163 genes responsive to T3 treatment and 187 genes differentially expressed between TRbetaPV/PV mice and wild-type littermates. Both the magnitude and gene make-up of the transcriptional response varied widely across tissues and conditions. We identified genes modulated in T3-dependent PV-independent, T3- and PV-dependent, and T3-independent PV-dependent pathways that illuminated the biological consequences of PV action in vivo. Most T3-responsive genes that were dysregulated in the heart and white adipose tissue of TRbetaPV/PV mice were repressed in T3-treated wild-type mice and upregulated in TRbetaPV/PV mice, suggesting the inappropriate activation of T3-suppressed genes in RTH. CONCLUSIONS: Comprehensive multi-tissue gene-expression analysis uncovered complex multiple signaling pathways that mediate the molecular actions of TRbeta mutants in vivo. In particular, the T3-independent mutant-dependent genomic response unveiled the contribution of a novel 'change-of-function' of TRbeta mutants to the pathogenesis of RTH. Thus, the molecular actions of TRbeta mutants are more complex than previously envisioned. PMID- 15128447 TI - Transcriptional regulation of protein complexes in yeast. AB - BACKGROUND: Multiprotein complexes play an essential role in many cellular processes. But our knowledge of the mechanism of their formation, regulation and lifetimes is very limited. We investigated transcriptional regulation of protein complexes in yeast using two approaches. First, known regulons, manually curated or identified by genome-wide screens, were mapped onto the components of multiprotein complexes. The complexes comprised manually curated ones and those characterized by high-throughput analyses. Second, putative regulatory sequence motifs were identified in the upstream regions of the genes involved in individual complexes and regulons were predicted on the basis of these motifs. RESULTS: Only a very small fraction of the analyzed complexes (5-6%) have subsets of their components mapping onto known regulons. Likewise, regulatory motifs are detected in only about 8-15% of the complexes, and in those, about half of the components are on average part of predicted regulons. In the manually curated complexes, the so-called 'permanent' assemblies have a larger fraction of their components belonging to putative regulons than 'transient' complexes. For the noisier set of complexes identified by high-throughput screens, valuable insights are obtained into the function and regulation of individual genes. CONCLUSIONS: A small fraction of the known multiprotein complexes in yeast seems to have at least a subset of their components co-regulated on the transcriptional level. Preliminary analysis of the regulatory motifs for these components suggests that the corresponding genes are likely to be co-regulated either together or in smaller subgroups, indicating that transcriptionally regulated modules might exist within complexes. PMID- 15128448 TI - Orthologous gene-expression profiling in multi-species models: search for candidate genes. AB - Microarray-driven gene-expression profiles are generally produced and analyzed for a single specific experimental model. We have assessed an analytical approach that simultaneously evaluates multi-species experimental models within a particular biological condition using orthologous genes as linkers for the various Affymetrix microarray platforms on multi-species models of ventilator associated lung injury. The results suggest that this approach may be a useful tool in the evaluation of biological processes of interest and selection of process-related candidate genes. PMID- 15128449 TI - Prolinks: a database of protein functional linkages derived from coevolution. AB - The advent of whole-genome sequencing has led to methods that infer protein function and linkages. We have combined four such algorithms (phylogenetic profile, Rosetta Stone, gene neighbor and gene cluster) in a single database- Prolinks--that spans 83 organisms and includes 10 million high-confidence links. The Proteome Navigator tool allows users to browse predicted linkage networks interactively, providing accompanying annotation from public databases. The Prolinks database and the Proteome Navigator tool are available for use online at http://dip.doe-mbi.ucla.edu/pronav. PMID- 15128450 TI - ELXR: a resource for rapid exon-directed sequence analysis. AB - ELXR (Exon Locator and Extractor for Resequencing) streamlines the process of determining exon/intron boundaries and designing PCR and sequencing primers for high-throughput resequencing of exons. We have pre-computed ELXR primer sets for all exons identified from the human, mouse, and rat mRNA reference sequence (RefSeq) public databases curated by the National Center for Biotechnology Information. The resulting exon-flanking PCR primer pairs have been compiled into a system called ELXRdb, which may be searched by keyword, gene name or RefSeq accession number. PMID- 15128451 TI - Genome2D: a visualization tool for the rapid analysis of bacterial transcriptome data. AB - Genome2D is a Windows-based software tool for visualization of bacterial transcriptome and customized datasets on linear chromosome maps constructed from annotated genome sequences. Genome2D facilitates the analysis of transcriptome data by using different color ranges to depict differences in gene-expression levels on a genome map. Such output format enables visual inspection of the transcriptome data, and will quickly reveal transcriptional units, without prior knowledge of expression level cutoff values. The compiled version of Genome2D is freely available for academic or non-profit use from http://molgen.biol.rug.nl/molgen/research/molgensoftware.php. PMID- 15128452 TI - Long-term clinical, immunologic and virologic impact of glucocorticoids on the chronic phase of HIV infection. AB - BACKGROUND: To test the hypothesis of down-regulating the increased immune system activation/destruction process associated with chronic HIV infection, we focused our interest on prednisolone (PDN), because we had showed that, in vitro, PDN had a strong anti-apoptotic activity on activated T cells of HIV-infected patients and no effect on viral replication. We thus designed in 1992 a pilot study to evaluate the clinical, immunologic and virologic effects of PDN. The drug was given to a group of 44 patients with CD4 T cells over 200/microl. After one year, no patient had developed clinical AIDS and the mean CD4 T cell count of the group had increased from 441 +/- 21 cells/microl to 553 +/- 43 cells/microl. Moreover, markers of immune activation had dropped back to normal levels while the mean viral load of the group had remained unchanged. Here we explore the long-term clinical, immunologic, and virologic impact of prednisolone on the chronic phase of HIV infection. METHODS: Retrospective study over 10 years starting between July 1992 and February 1993. A total of 44 patients with CD4 cells/microl ranging from 207 to 775 were treated with prednisolone, 0.5 mg/kg/d, over 6 months and 0.3 mg/kg/d thereafter. RESULTS: No clinical AIDS developed under prednisolone; side effects of the drug were mild. CD4 cells which increased from 421 cells/microl at entry to 625 cells/microl at day 15, slowly decreased to reach 426 cells/microl after two years; T cell apoptosis and activation markers dropped within 15 days to normal levels and reincreased slowly thereafter. Serum viral loads remained stable. The percentage of patients maintaining CD4 cells over entry was 43.2% at two years, 11.4% at five years and 4.6% at 10 years. Initial viral load was highly predictive of the rate of CD4 decrease under prednisolone. CONCLUSIONS: Prednisolone postponed CD4 cell decrease in a viral load dependent manner for a median of two years and for up to 10 years in a fraction of the patients with a low viral load. These findings might stimulate clinical trials as well as biological research on the role of antiapoptotic drugs in HIV infection. PMID- 15128453 TI - Multimodal treatment for resectable epithelial type malignant pleural mesothelioma. AB - BACKGROUND: Malignant pleural mesothelioma is a rare malignancy. The outcome remains poor despite complete surgical resection. PATIENTS AND METHODS: Eleven patients with histologicaly proven epithelial type malignant pleural mesothelioma undergoing extrapleural pneumonectomy with systemic chemotherapy and/or radiotherapy before and after surgical resection were retrospectively reviewed. RESULTS: Ten out of 11 patients underwent complete surgical resection, of these 7 patients had stage I disease. Of these 7 patients, 5 are alive without any recurrence, a 2-year survival rate of 80% was observed in this group. There was no operative mortality or morbidity. CONCLUSION: Extrapleural pneumonectomy with perioperative adjuvant treatment is safe and effective procedure for epithelial type malignant pleural mesothelioma. PMID- 15128454 TI - Markers of small cell lung cancer. AB - Lung cancer is the number one cause of cancer death; however, no specific serum biomarker is available till date for detection of early lung cancer. Despite good initial response to chemotherapy, small-cell lung cancer (SCLC) has a poor prognosis. Therefore, it is important to identify molecular markers that might influence survival and may serve as potential therapeutic targets. The review aims to summarize the current knowledge of serum biomarkers in SCLC to improve diagnostic efficiency in the detection of tumor progression in lung cancer. The current knowledge on the known serum cytokines and tumor biomarkers of SCLC is emphasized. Recent findings in the search for novel diagnostic and therapeutic molecular markers using the emerging genomic technology for detecting lung cancer are also described. It is believed that implementing these new research techniques will facilitate and improve early detection, prognostication and better treatment of SCLC. PMID- 15128455 TI - Women with coronary artery disease report worse health-related quality of life outcomes compared to men. AB - BACKGROUND: Although there have been substantial medical advances that improve the outcomes following cardiac ischemic events, gender differences in the treatment and course of recovery for patients with coronary artery disease (CAD) continue to exist. There is a general paucity of data comparing the health related quality of life (HRQOL) in men and women undergoing treatment for CAD. The purpose of this study was to compare HRQOL outcomes of men and women in Alberta, at one-year following initial catheterization, after adjustment for known demographic, co-morbid, and disease severity predictors of outcome. METHOD: The HRQOL outcome data were collected by means of a self-reported questionnaire mailed to patients on or near the one-year anniversary of their initial cardiac catheterization. Using the Seattle Angina Questionnaire (SAQ), 5 dimensions of HRQOL were measured: exertional capacity, anginal stability, anginal frequency, quality of life and treatment satisfaction. Data from the APPROACH registry were used to risk-adjust the SAQ scale scores. Two analytical strategies were used including general least squares linear modeling, and proportional odds modeling sometimes referred to as the "ordinal logistic modeling". RESULTS: 3392 (78.1%) patients responded to the follow-up survey. The adjusted proportional odds ratios for men relative to women (PORs > 1 = better) indicated that men reported significantly better HRQOL on all 5 SAQ dimensions as compared to women. (PORs: Exertional Capacity 3.38 (2.75-4.15), Anginal Stability 1.23 (1.03-1.47), Anginal Frequency 1.70 (1.43-2.01), Treatment Satisfaction 1.27 (1.07-1.50), and QOL 1.74 (1.48-2.04). CONCLUSIONS: Women with CAD consistently reported worse HRQOL at one year follow-up compared to men. These findings underline the fact that conclusions based on research performed on men with CAD may not be valid for women and that more gender-related research is needed. Future studies are needed to further examine gender differences in psychosocial adjustment following treatment for CAD, as adjustment for traditional clinical variables fails to explain sex differences in health related quality of life outcomes. PMID- 15128457 TI - Mutagenicity of the peroxisome proliferators clofibrate, Wyeth 14,643 and di-2 ethylhexyl phthalate in the lacZ plasmid-based transgenic mouse mutation assay. AB - BACKGROUND: Peroxisome proliferators are considered rodent carcinogens that are putative human non-carcinogens based on the presumed absence of direct genetic toxicity in rodent and human cells and the resistance of human cells to the induction of peroxisomes by peroxisome proliferators. The highly sensitive lacZ plasmid-based transgenic mouse mutation assay was employed to investigate the mutagenicity of several peroxisome proliferators based on several lines of evidence suggesting that these agents may in fact exert a genotoxic effect. METHODS: Male and female lacZ-plasmid based transgenic mice were treated at 4 months of age with 6 doses of 2,333 mg di-2-ethylhexyl phthalate (DHEP), 200 mg Wyeth-14,643, or 90 mg clofibrate per kg of bodyweight, respectively, over a two week period. Control animals were treated with the respective vehicles only (35% propyl glycol for DEHP and Wyeth-14,643 treatment controls and sterile water for clofibrate treatment controls).The mutant frequency in liver, kidney and spleen DNA was determined as the proportion of retrieved mutant and wild-type lacZ plasmids expressed in Escherichia Coli C host cells employing a positive selection system for mutant plasmids. RESULTS: Exposure to DEHP or Wyeth-14,643 significantly increased the mutant frequency in liver, but not in kidney or spleen, of both female and male mice. Treatment with clofibrate did not lead to an increased mutant frequency in any of the organs studied. CONCLUSION: The results indicate that some peroxisome proliferators display an organ-specific mutagenicity in lacZ plasmid-based transgenic mice consistent with historical observations of organ- and compound-specific carcinogenicity. PMID- 15128456 TI - Usefulness of EQ-5D in assessing health status in primary care patients with major depressive disorder. AB - OBJECTIVES: Major depressive disorder (MDD) is a prevalent psychiatric disorder associated with impaired patient functioning and reductions in health-related quality of life (HRQL). The present study describes the impact of MDD on patients' HRQL and examines preference-based health state differences by patient features and clinical characteristics. METHODS: 95 French primary care practitioners recruited 250 patients with a DSM-IV diagnosis of MDD for inclusion in an eight-week follow-up cohort. Patient assessments included the Montgomery Asberg Depression Rating Scale (MADRS), the Clinical Global Impression of Severity (CGI), the Short Form-36 Item scale (SF-36), the Quality of Life Depression Scale (QLDS) and the EuroQoL (EQ-5D). RESULTS: The mean EQ-5D utility at baseline was 0.33, and 8% of patients rated their health state as worse than death. There were no statistically significant differences in utilities by demographic features. Significant differences were found in mean utilities by level of disease severity assessed by CGI. The different clinical response profiles, assessed by MADRS, were also revealed by EQ-5D at endpoint: 0.85 for responders remitters, 0.72 for responders non-remitter, and 0.58 for non responders. Even if HRQL and EQ-5D were moderately correlated, they shared only 40% of variance between baseline and endpoint. CONCLUSIONS: Self-reported patient valuations for depression are important patient-reported outcomes for cost effectiveness evaluations of new antidepressant compounds and help in further understanding patient compliance with antidepressant treatment. PMID- 15128458 TI - Mutation at the position 2058 of the 23S rRNA as a cause of macrolide resistance in Streptococcus pyogenes. AB - BACKGROUND: In streptococci, three macrolide resistance determinants (erm(B), erm(TR) and mef(A)) have been found. In addition, certain mutations at the ribosomal 23S RNA can cause resistance to macrolides. Mutation at the position 2058 of the 23S rRNA of the Streptococcus pyogenes as a cause of macrolide resistance has not been described before. METHODS: Antibiotic resistance determinations for the clinical S. pyogenes strain ni4277 were done using the agar dilution technique. Macrolide resistance mechanisms were studied by PCR and sequencing. All six rRNA operons were amplified using operon-specific PCR. The PCR products were partially sequenced in order to resolve the sequences of different 23S rRNA genes. RESULTS: One clinical isolate of S. pyogenes carrying an adenine to guanine mutation at the position 2058 of the 23S rRNA in five of the six possible rRNA genes but having no other known macrolide resistance determinants is described. The strain was highly resistant to macrolides and azalides, having erythromycin and azithromycin MICs > 256 microgram/ml. It was resistant to lincosamides (clindamycin MIC 16 microgram/ml) and also MIC values for ketolides were clearly elevated. The MIC for telithromycin was 16 microgram/ml. CONCLUSION: In this clinical S. pyogenes strain, a mutation at the position 2058 was detected. No other macrolide resistance-causing determinants were detected. This mutation is known to cause macrolide resistance in other bacteria. We can conclude that this mutation was the most probable cause of macrolide, lincosamide and ketolide resistance in this strain. PMID- 15128459 TI - Air pollution attributable postneonatal infant mortality in U.S. metropolitan areas: a risk assessment study. AB - BACKGROUND: The impact of outdoor air pollution on infant mortality has not been quantified. METHODS: Based on exposure-response functions from a U.S. cohort study, we assessed the attributable risk of postneonatal infant mortality in 23 U.S. metropolitan areas related to particulate matter <10 microm in diameter (PM10) as a surrogate of total air pollution. RESULTS: The estimated proportion of all cause mortality, sudden infant death syndrome (normal birth weight infants only) and respiratory disease mortality (normal birth weight) attributable to PM10 above a chosen reference value of 12.0 microg/m3 PM10 was 6% (95% confidence interval 3-11%), 16% (95% confidence interval 9-23%) and 24% (95% confidence interval 7-44%), respectively. The expected number of infant deaths per year in the selected areas was 106 (95% confidence interval 53-185), 79 (95% confidence interval 46-111) and 15 (95% confidence interval 5-27), respectively. Approximately 75% of cases were from areas where the current levels are at or below the new U.S. PM2.5 standard of 15 microg/m3 (equivalent to 25 microg/m3 PM10). In a country where infant mortality rates and air pollution levels are relatively low, ambient air pollution as measured by particulate matter contributes to a substantial fraction of infant death, especially for those due to sudden infant death syndrome and respiratory disease. Even if all counties would comply to the new PM2.5 standard, the majority of the estimated burden would remain. CONCLUSION: Given the inherent limitations of risk assessments, further studies are needed to support and quantify the relationship between infant mortality and air pollution. PMID- 15128460 TI - The Oslo Health Study: The impact of self-selection in a large, population-based survey. AB - BACKGROUND: Research on health equity which mainly utilises population-based surveys, may be hampered by serious selection bias due to a considerable number of invitees declining to participate. Sufficient information from all the non responders is rarely available to quantify this bias. Predictors of attendance, magnitude and direction of non-response bias in prevalence estimates and association measures, are investigated based on information from all 40 888 invitees to the Oslo Health Study. METHODS: The analyses were based on linkage between public registers in Statistics Norway and the Oslo Health Study, a population-based survey conducted in 2000/2001 inviting all citizens aged 30, 40, 45, 59-60 and 75-76 years. Attendance was 46%. Weighted analyses, logistic regression and sensitivity analyses are performed to evaluate possible selection bias. RESULTS: The response rate was positively associated with age, educational attendance, total income, female gender, married, born in a Western county, living in the outer city residential regions and not receiving disability benefit. However, self-rated health, smoking, BMI and mental health (HCSL) in the attendees differed only slightly from estimated prevalence values in the target population when weighted by the inverse of the probability of attendance.Observed values differed only moderately provided that the non-attending individuals differed from those attending by no more than 50%. Even though persons receiving disability benefit had lower attendance, the associations between disability and education, residential region and marital status were found to be unbiased. The association between country of birth and disability benefit was somewhat more evident among attendees. CONCLUSIONS: Self-selection according to sociodemographic variables had little impact on prevalence estimates. As indicated by disability benefit, unhealthy persons attended to a lesser degree than healthy individuals, but social inequality in health by different sociodemographic variables seemed unbiased. If anything we would expect an overestimation of the odds ratio of chronic disease among persons born in non western countries. PMID- 15128461 TI - Spatial clustering of filarial transmission before and after a Mass Drug Administration in a setting of low infection prevalence. AB - BACKGROUND: In the global program for the elimination of lymphatic filariasis (LF) longitudinal assessment of the prevalence of microfilaremia and antigenemia is recommended to monitor the effect of mass treatment on transmission. Additional monitoring tools such as entomologic and antibody methods may be useful in identifying residual foci of infection. In this study, we characterized serologic markers of infection and exposure spatially both before and after mass treatment, in an area of initial low Wuchereria bancrofti infection prevalence. METHODS: Consenting persons in the sentinel community were tested for circulating microfilaria and antigen (by immunochromatographic test) before and after the 1st annual mass drug administration of diethylcarbamazine and albendazole. A cohort of 161 persons provided serum specimens both years that were tested for antifilarial IgG (1 and 4) antibody. Every house was mapped using a differential Global Positioning System; this information was linked to the serologic data. W. bancrofti infection in the mosquito vector was assessed with year-round collection. Multiple linear regression was used to investigate the influence of antigen-positive persons on the antifilarial antibody responses of antigen negative neighbors. RESULTS: After mass treatment, decreases were observed in the sentinel site in the overall prevalence of antigen (10.4% to 6.3%) and microfilaremia (0.9 to 0.4%). Of the persons in the cohort that provided serum specimens both years, 79% received treatment. Antigen prevalence decreased from 15.0% to 8.7%. Among 126 persons who received treatment, antigen and antifilarial IgG1 prevalence decreased significantly (p = 0.002 and 0.001, respectively). Among 34 persons who did not receive treatment, antifilarial IgG1 prevalence increased significantly (p = 0.003). Average antifilarial IgG1 levels decreased in households with high treatment coverage and increased in households that refused treatment. Each 10-meter increase in distance from the residence of a person who was antigen-positive in 2000 was associated a 4.68 unit decrease in antifilarial IgG1 level in 2001, controlling for other factors (p = 0.04). DISCUSSION: Antifilarial antibody assays can be used as a measure of filarial exposure. Our results suggest that micro-scale spatial heterogeneity exists in LF exposure and infection. Treatment appeared to be associated with reduced exposure at the sub-community level, suggesting the need to achieve high and homogeneous coverage. Public health messages should note the benefits of having one's neighbors receive treatment with antifilarial drugs. PMID- 15128462 TI - Examination of NRCAM, LRRN3, KIAA0716, and LAMB1 as autism candidate genes. AB - BACKGROUND: A substantial body of research supports a genetic involvement in autism. Furthermore, results from various genomic screens implicate a region on chromosome 7q31 as harboring an autism susceptibility variant. We previously narrowed this 34 cM region to a 3 cM critical region (located between D7S496 and D7S2418) using the Collaborative Linkage Study of Autism (CLSA) chromosome 7 linked families. This interval encompasses about 4.5 Mb of genomic DNA and encodes over fifty known and predicted genes. Four candidate genes (NRCAM, LRRN3, KIAA0716, and LAMB1) in this region were chosen for examination based on their proximity to the marker most consistently cosegregating with autism in these families (D7S1817), their tissue expression patterns, and likely biological relevance to autism. METHODS: Thirty-six intronic and exonic single nucleotide polymorphisms (SNPs) and one microsatellite marker within and around these four candidate genes were genotyped in 30 chromosome 7q31 linked families. Multiple SNPs were used to provide as complete coverage as possible since linkage disequilibrium can vary dramatically across even very short distances within a gene. Analyses of these data used the Pedigree Disequilibrium Test for single markers and a multilocus likelihood ratio test. RESULTS: As expected, linkage disequilibrium occurred within each of these genes but we did not observe significant LD across genes. None of the polymorphisms in NRCAM, LRRN3, or KIAA0716 gave p < 0.05 suggesting that none of these genes is associated with autism susceptibility in this subset of chromosome 7-linked families. However, with LAMB1, the allelic association analysis revealed suggestive evidence for a positive association, including one individual SNP (p = 0.02) and three separate two-SNP haplotypes across the gene (p = 0.007, 0.012, and 0.012). CONCLUSIONS: NRCAM, LRRN3, KIAA0716 are unlikely to be involved in autism. There is some evidence that variation in or near the LAMB1 gene may be involved in autism. PMID- 15128463 TI - A randomized trial to assess the impact of an antithrombotic decision aid in patients with nonvalvular atrial fibrillation: the DAAFI trial protocol [ISRCTN14429643]. AB - BACKGROUND: Decision aids are often advocated as a means to assist patient and health care provider decision making when faced with complicated treatment or screening decisions. Despite an exponential growth in the availability of decision aids in recent years, their impact on long-term treatment decisions and patient adherence is uncertain due to a paucity of rigorous studies. The choice of antithrombotic therapy for nonvalvular atrial fibrillation (NVAF) is one condition for which a trade-off exists between the potential risks and benefits of competing therapies, and the need to involve patients in decision making has been clearly identified. This study will evaluate whether an evidence-based patient decision aid for patients with NVAF can improve the appropriateness of antithrombotic therapy use by patients and their family physicians. DESIGN: A multi-center, two-armed cluster randomized trial based in community family practices in which patients with NVAF will be randomized to decision aid or usual care. Patients will receive one of four decision aids depending on their baseline stroke risk. The primary outcome is the provision of "appropriate antithrombotic therapy" at 3 months to study participants (appropriateness defined as per the 2001 American College of Chest Physicians recommendations for NVAF). In addition, the impact of this decision aid on patient knowledge, decisional conflict, well being, and adherence will be assessed after 3 months, 6 months, and 12 months. PMID- 15128464 TI - Comparison of 16S rRNA gene sequences of genus Methanobrevibacter. AB - BACKGROUND: The phylogeny of the genus Methanobrevibacter was established almost 25 years ago on the basis of the similarities of the 16S rRNA oligonucleotide catalogs. Since then, many 16S rRNA gene sequences of newly isolated strains or clones representing the genus Methanobrevibacter have been deposited. We tried to reorganize the 16S rRNA gene sequences of this genus and revise the taxonomic affiliation of the isolates and clones representing the genus Methanobrevibacter. RESULTS: The phylogenetic analysis of the genus based on 786 bp aligned region from fifty-four representative sequences of the 120 available sequences for the genus revealed seven multi-member groups namely, Ruminantium, Smithii, Woesei, Curvatus, Arboriphilicus, Filiformis, and the Termite gut symbionts along with three separate lineages represented by Mbr. wolinii, Mbr. acididurans, and termite gut flagellate symbiont LHD12. The cophenetic correlation coefficient, a test for the ultrametric properties of the 16S rRNA gene sequences used for the tree was found to be 0.913 indicating the high degree of goodness of fit of the tree topology. A significant relationship was found between the 16S rRNA sequence similarity (S) and the extent of DNA hybridization (D) for the genus with the correlation coefficient (r) for logD and logS, and for [ln(-lnD) and ln(-lnS)] being 0.73 and 0.796 respectively. Our analysis revealed that for this genus, when S = 0.984, D would be <70% at least 99% of the times, and with 70% D as the species "cutoff", any 16S rRNA gene sequence showing <98% sequence similarity can be considered as a separate species. In addition, we deduced group specific signature positions that have remained conserved in evolution of the genus. CONCLUSIONS: A very significant relationship between D and S was found to exist for the genus Methanobrevibacter, implying that it is possible to predict D from S with a known precision for the genus. We propose to include the termite gut flagellate symbiont LHD12, the methanogenic endosymbionts of the ciliate Nyctotherus ovalis, and rat feces isolate RT reported earlier, as separate species of the genus Methanobrevibacter. PMID- 15128465 TI - Calcium antagonist strategies for coronary artery disease. PMID- 15128466 TI - Diastolic heart failure: the forgotten manifestation of hypertensive heart disease. AB - Heart failure (HF) is a progressively debilitating disorder characterized by frequent hospital admissions and high annual mortality rates. Coronary artery disease (CAD), hypertension, and aging are major risk factors for the development/progression of HF. For years, most of the attention has been focused on HF caused by reduced left ventricular (LV) systolic function, largely attributable to CAD. It is now generally accepted that nearly 50% of elderly patients with HF might have normal or preserved LV systolic function. This condition is commonly referred to as a distinct type of HF caused by LV diastolic dysfunction, and it often accompanies hypertensive heart disease. Isolated diastolic HF is increasingly recognized as the dominant cause of symptoms and hospitalizations from HF in a large proportion of individuals aged 65 and older. However, the clinicians caring for patients with diastolic HF do not fully understand its cause, how it progresses, or how it could be appropriately diagnosed and treated. Because varying degrees of systolic and diastolic dysfunction might coexist in any individual patient, and given the limitation of current diagnostic tools, the overall impact of isolated diastolic HF continues to evolve. Ongoing clinical trials are testing new strategies for treatment of diastolic HF. PMID- 15128467 TI - Vascular abnormalities in hypertension: cause, effect, or therapeutic target? AB - Alteration in the physical properties of arterial blood vessels act as the substrate for end-organ damage and the occurrence of vascular events in hypertension. Structural and functional alteration in the microcirculation represents a prevalent and characteristic abnormality described in the earliest stages of hypertension. In addition to lowering arterial pressure, the importance of reversing structural abnormalities and restoring endothelial function has recently been emphasized. The normalization of microvascular function does not always correlate with blood pressure reduction and might depend on the class of antihypertensive agent employed. Because altered resistance vessel structure and function is a recognized hallmark of hypertension, institution of drug therapy that corrects the associated pathogenic vascular abnormalities is a rational, if unproven, clinical goal. PMID- 15128468 TI - Is proteinuria a plausible target of therapy? AB - Microalbuminuria is an independent marker of cardiovascular risk, irrespective of kidney disease. Recent pharmacologic interventions have resulted in a significant delay and even an arrest in the progression of microalbuminuria to macroalbuminuria or to chronic kidney disease. Focus should be placed on agents that not only lower blood pressure but also improve albuminuria levels. Current guidelines recommend that hypertensive patients with renal disease should be started on agents that block the renin-angiotensin-aldosterone system, such as angiotensin-converting enzyme (ACE) inhibitors or angiotensin receptor blockers (ARBs). The use of three to four different agents is frequently necessary to reach the guideline goal blood pressure of 130/80 mm Hg. PMID- 15128469 TI - Is the angiotensin II Type 2 receptor cerebroprotective? AB - Most of the deleterious effects of angiotensin II (Ang II) on blood pressure (BP), cardiovascular remodeling, and atherosclerosis are mediated by Ang II type 1 (AT1)-receptor activation. This explains why Ang-II-decreasing or blocking drugs have been successful in decreasing global cardiovascular morbimortality in patients with cardiac complications. However, in primary or secondary stroke prevention trials in patients with low cardiac risk, b-blockers and angiotensin converting enzyme inhibitors (ACEIs), which decrease Ang II formation, seem to be less protective than thiazides and dihydropyridines, which increase Ang II. When compared with a beta-blocker, an Ang II-increasing AT1-receptor blocker better protects against stroke but not against cardiac events, whereas an ACEI gives the same protection against both cardiac and cerebral events. This dissociation between blood-pressure-independent cardiac and cerebral protection between b blockers or ACEIs versus AT1-blockers in patients with low cardiac risk can be best explained if, besides the beneficial vascular effect of AT1-receptor blunting, there is evidence of a beneficial effect of non-AT1-receptor activation. In this review, we present experimental evidence for AT2- and AT4 receptor-mediated brain-anti-ischemic mechanisms and propose a direct comparison of AT1-blockers with ACEIs to prove the clinical effectiveness of non-AT1 mediated mechanisms in stroke prevention, particularly in patients with a higher risk for stroke than for cardiac complications. PMID- 15128470 TI - Pulse pressure, heart rate, and drug treatment of hypertension. AB - Peak-systolic blood pressure (SBP) and end-diastolic blood pressure (DBP) have been considered the exclusive mechanical factors predicting cardiovascular (CV) risk in populations of normotensive and hypertensive subjects. The purpose of this review is to show that in addition to SBP and DBP, other hemodynamic indices that have particular relevance for coronary complications and originate from pulse pressure (PP) should be taken into account, namely brachial PP and, potentially, heart rate (HR). In normotensive and hypertensive populations, increased PP is an independent predictor for myocardial infarction, more powerful than SBP, even in hypertensive subjects under successful antihypertensive drug therapy. Increased HR is an additional CV risk factor, acting particularly through the presence of an enhanced PP x HR product. Such findings, observed during a period in which standard antihypertensive agents are known to reduce SBP and PP to a much lesser extent than DBP, require the development of intervention trials using drugs acting specifically on SBP, PP, and arterial stiffness. PMID- 15128471 TI - Aldosterone and aldosterone antagonism in systemic hypertension. AB - Aldosterone mediates both water and electrolyte balance by acting on the renal mineralocorticoid receptors. Recent experimental studies have also documented the presence of these receptors in other body organs, including the brain, blood vessels, and heart, suggesting that aldosterone plays a larger role in normal physiologic function and in cardiovascular diseases such as systemic hypertension and congestive heart failure (CHF). The nonspecific aldosterone inhibitor spironolactone, and the selective aldosterone inhibitor eplerenone, are both approved for clinical use in treating patients with hypertension and/or symptomatic CHF. Studies have shown that spironolactone lowers blood pressure, improves endothelial function, reduces myocardial hypertrophy and fibrosis, and lowers the incidence of fatal arrhythmias. Eplerenone, which is more specific for the mineralocorticoid receptor, appears to provide all the beneficial effects of spironolactone in hypertensive patients, with the potential to modify many of the side effects related to nonspecific steroid-receptor blockade. Hyperkalemia remains a potential problem with all aldosterone antagonists. PMID- 15128472 TI - Prevention of dementia and cerebroprotection with antihypertensive drugs. AB - High blood pressure is a major risk factor for stroke and is also closely correlated with cognitive decline and dementia. Indeed, most longitudinal studies showed that cognitive functioning is often inversely proportional to blood pressure values measured 15 or 20 years previously. Because of the aging of the population, the frequency of stroke and dementia will dramatically increase in the coming years. Therefore, the prevention of cerebrovascular and cognitive disorders represents a major challenge. Antihypertensive drugs have shown clinical benefits in both primary and secondary prevention of strokes. Consensus is generally that blood-pressure lowering represents the major determinant of the benefit conferred by the antihypertensive treatment for stroke prevention; however, recent studies have suggested some differences between classes of antihypertensive drugs. The results of therapeutic trials (Systolic Hypertension in Europe, Perindopril Protection Against Recurrent Stroke Study [PROGRESS]) open the way to the prevention of dementia (vascular or Alzheimer's type) by antihypertensive treatments. These two studies suggest different mechanisms for the prevention of cognitive decline using antihypertensive drugs. In this context, reduced incidence of dementia should be the primary outcome of future trials comparing different classes of antihypertensive drugs. PMID- 15128473 TI - Adding low-dose spironolactone to multidrug regimens for resistant hypertension. PMID- 15128474 TI - The Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack (ALLHAT) Trial: focus on the diabetic patient. PMID- 15128475 TI - Diabetes, hypertension, and cardiovascular derangements: pathophysiology and management. AB - Hypertension frequently coexists with diabetes mellitus, occurring twice as frequently in diabetic as in nondiabetic persons. It accounts for up to 75% of added cardiovascular disease (CVD) risk in people with diabetes, contributing significantly to the overall morbidity and mortality in this high-risk population. Patients with hypertension are two times more prone to have diabetes than are normotensive persons. Hypertension substantially increases the risk for coronary heart disease (CHD), stroke, retinopathy, and nephropathy. In patients with type 2 diabetes, hypertension usually clusters with the other components of the cardiometabolic syndrome, such as microalbuminuria, central obesity, insulin resistance, dyslipidemia, hypercoagulation, increased inflammation, and left ventricular hypertrophy (LVH). In type 1 diabetes, hypertension often occurs subsequent to the development of diabetic nephropathy. Hypertension in people with diabetes is characterized by volume expansion, increased salt sensitivity, isolated systolic blood pressure (BP) elevation, loss of the nocturnal dipping of BP and pulse, and increased propensity toward orthostatic hypotension and albuminuria. Among the treatment strategies tested in hypertensive diabetic persons, low-density lipoprotein (LDL)-cholesterol lowering to less than 100 mg/dL and aggressive BP control to less than 130/80 mm Hg have proven effective in CVD risk reduction. The combination of two or more drugs is usually necessary to achieve the target BP. PMID- 15128476 TI - Adrenocortical hypertension. AB - Primary aldosteronism, congenital adrenal hyperplasia, Cushing's syndrome, glucocorticoid-remediable aldosteronism, and corticotropin-dependent forms of adrenal pathology can cause hypertension by excessive production of adrenocortical hormones. Although traditional biochemical assays continue to be used, genetic testing has simplified the diagnosis of glucocorticoid-remediable aldosteronism. Also, new interventional radiologic approaches for the diagnosis and treatment of corticotropin-dependent forms of Cushing's syndrome are available. Medical and surgical approaches, however, still remain viable options for treatment. PMID- 15128477 TI - Obesity-related hypertension: is there a role for selective leptin resistance? AB - Obesity is a risk factor for cardiovascular diseases, in particular for hypertension. Serum leptin levels and sympathetic nerve activity are both increased in obesity. Leptin has been demonstrated to increase sympathetic nerve activity. Thus, leptin-dependent sympathoactivation might contribute to obesity related hypertension. However, leptin resistance occurs in obesity. One possibility is that leptin resistance is selective to the metabolic effects of leptin, sparing its sympathoexcitatory actions. In this article, we review experimental evidence supporting the novel concept of selective leptin resistance. We also discuss the sympathetic actions of leptin that are relevant to blood pressure modulation and potential mechanisms of leptin resistance. Disruption of leptin intracellular signaling pathways and resistance of specific leptin-responsive neural networks provide theoretic models of selective leptin resistance. However, most information about leptin-sympathetic actions and leptin resistance mechanisms derive from in vitro and animal studies. Future research in humans is widely awaited. PMID- 15128478 TI - Leptin, sympathetic nervous system, and baroreflex function. AB - In addition to its direct effects on energy metabolism and caloric intake, leptin exerts several circulatory effects that appear to be mediated by an interaction with the sympathetic nervous system and the major reflexogenic area involved in cardiovascular homeostatic control--that is, the arterial baroreflex. In this paper, the relationships between the adipocyte hormone and the neuroadrenergic function are reviewed, taking into account data collected in experimental animal models as well as in human cardiovascular (hypertension and heart failure) and noncardiovascular (obesity) diseases that are characterized by a hyperadrenergic state coupled with a hyperleptinemia. PMID- 15128479 TI - The global epidemic of obesity: are we becoming more sympathetic? AB - The relationship between obesity and sympathetic nervous system (SNS) behavior has been controversial. Existing paradigms have been opposed in their views of the directional changes in SNS activity in obesity. In addition, limitations of previous approaches to assess SNS activity have produced inconsistent findings. However, the use of state-of-the-art neurochemical and neurophysiologic techniques has improved our current understanding of this issue. There is regional heterogeneity in the SNS activation associated with obesity, with the renal and skeletal muscle circulations, but not the heart, being targets for SNS activation. Abdominal visceral fat appears to be an important depot linking obesity and skeletal muscle SNS activation. The impact of this depot on SNS activity to the kidney or other regions is unknown. Future studies are needed to address this issue as well as the mechanisms and consequences of SNS activation in visceral obesity. PMID- 15128480 TI - Enterotoxins, enteric nerves, and intestinal secretion. PMID- 15128481 TI - Treatment options for eosinophilic esophagitis: elemental diet. PMID- 15128482 TI - Treatment options for eosinophilic esophagitis: montelukast. PMID- 15128483 TI - The implications of Helicobacter pylori infection for gastroesophageal reflux disease: studies presented at Digestive Disease Week 2003. AB - The relationship between Helicobacter pylori infection and gastroesophageal reflux disease (GERD) remains controversial. Over 20 studies presented at the 2003 Digestive Disease Week meeting examined the implications of H. pylori infection for GERD. Although the findings were not uniform, most of the studies presented indicated a negative association between the presence of H. pylori and the presence of GERD symptoms, signs, or complications. In addition, eradication of H. pylori infection was related to the emergence of GERD and its complications. However, most studies favored a role for H. pylori rather than GERD in the development of pathologic changes in the gastric cardia. Finally, a few studies suggested the intriguing possibility that the modulation of GERD by H. pylori could be through the effects of H. pylori eradication on weight gain. PMID- 15128484 TI - Endoscopic techniques for treatment of gastroesophageal reflux disease: a review of abstracts from Digestive Disease Week 2003. AB - Endoscopic treatments of gastroesophageal reflux disease (GERD) are a new and potentially useful form of therapy for reflux disorders. This type of treatment was first introduced at Digestive Disease Week (DDW) in 2000, and the most recent DDW, in 2003, included a growing number of abstracts describing further experience with endoscopic GERD treatments. The studies are still generally open label trials involving relatively small patient groups given a variety of unique endoscopic GERD treatments, including radiofrequency, thermal ablation, plication suturing, and polymer injection and implantation techniques. Longer patient follow-up after Stretta (Curon Medical, Sunnyvale, CA) procedures, the permanency rate of EndoCinch (Bard Interventional Products, Billerica, MA) plication sutures, and new experiences with Enteryx (Boston Scientific, Natick, MA) and Gatekeeper (Medtronic, Minneapolis, MN) procedures were highlighted at DDW 2003. Despite these unique devices with significant potential for treatment of GERD, caution was expressed concerning their clinical use in anything other than appropriately designed, scientifically validated clinical trials. PMID- 15128485 TI - Barrett's esophagus: best of Digestive Disease Week 2003. AB - The definition of Barrett's esophagus is conceptually simple. It is the replacement of the normal squamous mucosa by specialized intestinal metaplasia within the esophagus. Barrett's esophagus would only be a clinical curiosity if not for its predisposition toward neoplastic change. In spite of all the knowledge and data gathered thus far on Barrett's, much remains unknown. Why do some patients develop Barrett's while others do not? Why do some with Barrett's advance to adenocarcinoma? Once Barrett's develops, what is the optimal prevention strategy for adenocarcinoma? These clinical unknowns touch only the tip of the iceberg and have made Barrett's esophagus a subject of intense research. This review offers some of the highlights from Digestive Disease Week 2003, comprised of posters of distinction and oral presentations. PMID- 15128486 TI - Management of esophageal varices: an update from Digestive Disease Week and American Association for the Study of Liver Diseases 2003. AB - Esophageal varices are a commonly encountered complication of cirrhosis. beta blockers are a well-established cornerstone of the treatment of portal hypertension and primary and secondary prophylaxis for prevention of esophageal variceal bleeding. However, not all patients tolerate this type of therapy. Moreover, the exact role of esophageal variceal band ligation alone or in combination with beta-blocker therapy in the management of patients with esophageal varices remains to be defined. This summary report presents a number of recent studies addressing these important issues. PMID- 15128487 TI - Wireless capsule endoscopy in the evaluation of the esophagus. AB - Wireless capsule endoscopy is an exciting and innovative technology that allows direct visualization of areas of the small bowel that were previously inaccessible to the "umbilicated" endoscope except during intraoperative enteroscopy. The wireless capsule endoscope permits noninvasive small bowel endoscopy in the ambulatory setting and has been proven to be a significant advance in the evaluation of patients with challenging small intestinal disease. PMID- 15128488 TI - Gastroesophageal reflux disease and baclofen: is there a light at the end of the tunnel? AB - Transient lower esophageal sphincter relaxations (TLESRs) are rapid and prolonged relaxations of the lower esophageal sphincter (LES) that are not associated with swallowing. They are the mechanism by which most gastroesophageal reflux episodes occur in normal people and in patients with esophagitis. Transient LES relaxations appear to be mediated by a vagovagal reflex initiated by gastric distention. Baclofen is a g-aminobutyric acid (GABA) derivative that inhibits the production of TLESRs by acting as a GABA(B) receptor agonist at one or more loci along the vagovagal reflex arc. Animal and human studies suggest that baclofen decreases the number of reflux events and amount of esophageal acid exposure. Baclofen or another GABA(B) receptor agonist may be clinically useful in treatment of gastroesophageal reflux disease. PMID- 15128489 TI - Zinc with oral rehydration for diarrhea in hospitalized children. PMID- 15128490 TI - Autoimmune hepatitis in children. AB - Autoimmune hepatitis (AIH) remains an enigmatic condition that affects children of all ages. Although much knowledge has emerged over the past four decades regarding autoimmune diseases, the lack of a spontaneous animal model has hindered a more complete understanding of the pathophysiology of AIH. Serum autoimmune markers, combined with biochemical and histologic evidence of hepatocellular injury and absent other diagnostic considerations, are necessary to diagnose AIH. The clinical spectrum ranges from asymptomatic elevation of aminotransferase levels to acute liver failure. Patients with AIH type 1 are more likely to be older and have cirrhosis at the time of diagnosis, whereas those with AIH type 2 may have a more fulminant course and require life-long immunosuppressant medications. Overlap syndromes occur in children but are rare. Treatment with prednisone and azathioprine, alone or in combination, reverses the clinical course and improves biochemical abnormalities. Should liver transplantation be necessary, autoimmune injury may recur in the allograft. PMID- 15128491 TI - Hepatobiliary complications of cystic fibrosis. AB - Cystic fibrosis (CF) is the most common potentially lethal genetic disease in the Caucasian population. The disease results from mutations in the gene for the cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated Cl( ) channel in the apical membrane of most secretory epithelia. In the liver, CFTR is located in biliary epithelial cells or cholangiocytes and gallbladder epithelia, where it appears to play a role in normal bile formation. However, how a defective CFTR protein leads to associated liver and biliary disease in a subset of patients with CF is unknown. Improvements in life expectancy have led to an increasing recognition of hepatobiliary complications from CF. Whereas the biliary tract disease is usually clinically evident, the liver involvement may progress silently, only manifesting as end-stage liver disease and portal hypertension. Unlike the pancreatic involvement in CF, a genotype-phenotype correlation is not apparent in the expression of liver disease, suggesting the presence of as yet unidentifiable "genetic modifiers" influencing disease expression. This review focuses on the pathogenesis, clinical manifestations, screening, diagnosis, and treatment of CF hepatobiliary disease. PMID- 15128492 TI - Pancreatitis in childhood. AB - Inflammatory disease of the pancreas falls into two major classifications: acute and chronic. Acute pancreatitis is a reversible process, whereas chronic pancreatitis produces irreversible changes in the architecture and function of the pancreas. The recent finding that mutations in the gene encoding cationic trypsinogen are associated with hereditary pancreatitis, the identification of genes that increase the risk for developing chronic pancreatitis, and advances in cell biology have contributed greatly to our understanding of the molecular mechanisms leading to pancreatitis. Although pancreatitis is less common in children than in adults, it still occurs with regularity and should be considered in any child with acute or chronic abdominal pain. The major difference between pancreatitis in children and adults lies in the etiologies and outcome of acute pancreatitis and in the etiology of chronic pancreatitis. The treatment of acute and chronic pancreatitis is similar at all ages. PMID- 15128493 TI - Across the developmental continuum of irritable bowel syndrome: clinical and pathophysiologic considerations. AB - Irritable bowel syndrome is a common functional gastrointestinal disorder that affects children and adults. The lack of consensus diagnostic criteria and pathophysiologic understanding has hampered clinical progress in diagnosing and treating this disorder. The recent development of the Rome diagnostic criteria, mapping of brain-gut pathways using neuroimaging, and serotonergic pharmacology have greatly advanced the field. Chronic and acute life stress, especially during childhood, has been recognized as central to the initiation of the disorder and the induction of acute symptoms. We propose a developmental continuum whereby the clinical presentation of irritable bowel syndrome changes with age from irritability during infancy, to diarrhea in toddlers, to recurring abdominal pain during school age, and to pain and altered bowel habits during later adolescence and adulthood. PMID- 15128494 TI - Novel foods to treat food allergy and gastrointestinal infection. AB - The gastrointestinal tract communicates directly with the external environment. Necessary nutrients must be absorbed and commensal bacteria tolerated, and foreign proteins, antigens, and pathogens must be simultaneously excluded or destroyed. Immaturity or disruption of the mucosal immune defenses increases vulnerability to food allergy, intolerance, and infectious disease. Diseases resulting from ingested foreign proteins and organisms are increasing and cause morbidity and mortality worldwide. There is no specific treatment for food allergy other than avoidance. Vaccination for infectious disease is limited by the cost and logistics of distribution and administration, particularly in developing countries. Novel strategies are being explored to modulate the gut mucosal immune system by altering protein expression in food. Crops are being developed to remove deleterious allergens to prevent immunogenic exposure while preserving nutritional quality. Local food plants that express protein fragments of pathogens might provide an effective means to stimulate gut mucosal immunity while increasing vaccine accessibility. PMID- 15128501 TI - The calcium-dependent activity of large-conductance, calcium-activated K+ channels is enhanced by Pyk2- and Hck-induced tyrosine phosphorylation. AB - Recent results showing that large-conductance, calcium-activated K(+) (BK(Ca)) channels undergo direct tyrosine phosphorylation in the presence of c-Src tyrosine kinase have suggested the involvement of these channels in Src-mediated signaling pathways. Given the important role for c-Src in integrin-mediated signal transduction, we have examined the potential regulation of BK(Ca) channels by proline-rich tyrosine kinase 2 (Pyk2), a calcium-sensitive tyrosine kinase activated upon integrin stimulation. Transient coexpression of murine BK(Ca) channels with either wild-type Pyk2 or hematopoietic cell kinase (Hck), a Src family kinase, led to an enhancement of BK(Ca) channel activity over the range of 1-10 microM free calcium, whereas coexpression with catalytically inactive forms of either kinase did not significantly alter BK(Ca) gating compared with channels expressed alone. In the presence of either wild-type Pyk2 or Hck, BK(Ca) alpha subunits were found to undergo tyrosine phosphorylation, as determined by immunoprecipitation and Western blotting strategies. However, tyrosine phosphorylation of the BK(Ca) alpha-subunit was not detected for channels expressed alone or together with inactive forms of either Pyk2 or Hck. Interestingly, wild-type, but not inactive, Pyk2 was also present in BK(Ca) channel immunoprecipitates, suggesting that Pyk2 may coassociate with the BK(Ca) channel complex after phosphorylation. Collectively, the observed modulation and phosphorylation of BK(Ca) channels by Pyk2 and a Src-family kinase may reflect a general cellular mechanism by which G protein-coupled receptor and/or integrin activation leads to the regulation of membrane ion channels. PMID- 15128502 TI - Fiber type and temperature dependence of inorganic phosphate: implications for fatigue. AB - Elevated levels of P(i) are thought to cause a substantial proportion of the loss in muscular force and power output during fatigue from intense contractile activity. However, support for this hypothesis is based, in part, on data from skinned single fibers obtained at low temperatures (< or =15 degrees C). The effect of high (30 mM) P(i) concentration on the contractile function of chemically skinned single fibers was examined at both low (15 degrees C) and high (30 degrees C) temperatures using fibers isolated from rat soleus (type I fibers) and gastrocnemius (type II fibers) muscles. Elevating P(i) from 0 to 30 mM at saturating free Ca(2+) levels depressed maximum isometric force (P(o)) by 54% at 15 degrees C and by 19% at 30 degrees C (P < 0.05; significant interaction) in type I fibers. Similarly, the P(o) of type II fibers was significantly more sensitive to high levels of P(i) at the lower (50% decrease) vs. higher temperature (5% decrease). The maximal shortening velocity of both type I and type II fibers was not significantly affected by elevated P(i) at either temperature. However, peak fiber power was depressed by 49% at 15 degrees C but by only 16% at 30 degrees C in type I fibers. Similarly, in type II fibers, peak power was depressed by 40 and 18% at 15 and 30 degrees C, respectively. These data suggest that near physiological temperatures and at saturating levels of intracellular Ca(2+), elevated levels of P(i) contribute less to fatigue than might be inferred from data obtained at lower temperatures. PMID- 15128503 TI - The 1.4-MDa apoptosome is a critical intermediate in apoptosome maturation. AB - Previously, we demonstrated that both 150 mM KCl and alkaline pH inhibit cytochrome c-mediated activation of procaspase-3 in a unique manner. To determine the mechanism of inhibition, we analyzed the effect of KCl and alkaline pH on the formation of apoptosomes (a large complex consisting of cytochrome c, Apaf-1, and procaspase-9/caspase-9) in vitro. Our results suggest that an initial approximately 700-kDa apoptosome matures through a 1.4-MDa intermediate before a approximately 700-kDa apoptosome is reformed and procaspase-3 is activated. We further demonstrate that 150 mM KCl interferes with the conversion of the initial approximately 700-kDa apoptosome to the 1.4-MDa intermediate, while alkaline pH "traps" the apoptosome in the 1.4-MDa intermediate. Analysis of the cleaved state of procaspase-9 and procaspase-3 suggests that the 1.4-MDa intermediate may be required for cleavage of procaspase-9. Consistent with these results, in vivo data suggest that blocking acidification during the induction of apoptosis inhibits activation of procaspase-3. On the basis of these results, we propose a model of apoptosome maturation. PMID- 15128504 TI - Distinct kinases are involved in contraction of cat esophageal and lower esophageal sphincter smooth muscles. AB - Contraction of smooth muscle depends on the balance of myosin light chain kinase (MLCK) and myosin light chain phosphatase (MLCP) activities. Because MLCK activation depends on the activation of calmodulin, which requires a high Ca(2+) concentration, phosphatase inhibition has been invoked to explain contraction at low cytosolic Ca(2+) levels. The link between activation of the Ca(2+) independent protein kinase Cepsilon (PKCepsilon) and MLC phosphorylation observed in the esophagus (ESO) (Sohn UD, Cao W, Tang DC, Stull JT, Haeberle JR, Wang CLA, Harnett KM, Behar J, and Biancani P. Am J Physiol Gastrointest Liver Physiol 281: G467-G478, 2001), however, has not been elucidated. We used phosphatase and kinase inhibitors and antibodies to signaling enzymes in combination with intact and saponin-permeabilized isolated smooth muscle cells from ESO and lower esophageal sphincter (LES) to examine PKCepsilon-dependent, Ca(2+)-independent signaling in ESO. The phosphatase inhibitors okadaic acid and microcystin-LR, as well as an antibody to the catalytic subunit of type 1 protein serine/threonine phosphatase, elicited similar contractions in ESO and LES. MLCK inhibitors (ML-7, ML-9, and SM-1) and antibodies to MLCK inhibited contraction induced by phosphatase inhibition in LES but not in ESO. The PKC inhibitor chelerythrine and antibodies to PKCepsilon, but not antibodies to PKCbetaII, inhibited contraction of ESO but not of LES. In ESO, okadaic acid triggered translocation of PKCepsilon from cytosolic to particulate fraction and increased activity of integrin-linked kinase (ILK). Antibodies to the mitogen-activated protein (MAP) kinases ERK1/ERK2 and to ILK, and the MAP kinase kinase (MEK) inhibitor PD-98059, inhibited okadaic acid-induced ILK activity and contraction of ESO. We conclude that phosphatase inhibition potentiates the effects of MLCK in LES but not in ESO. Contraction of ESO is mediated by activation of PKCepsilon, MEK, ERK1/2, and ILK. PMID- 15128505 TI - Characterization of humanized antibodies secreted by Aspergillus niger. AB - Two different humanized immunoglobulin G1(kappa) antibodies and an Fab' fragment were produced by Aspergillus niger. The antibodies were secreted into the culture supernatant. Both light and heavy chains were initially synthesized as fusion proteins with native glucoamylase. After antibody assembly, cleavage by A. niger KexB protease allowed the release of free antibody. Purification by hydrophobic charge induction chromatography proved effective at removing any antibody to which glucoamylase remained attached. Glycosylation at N297 in the Fc region of the heavy chain was observed, but this site was unoccupied on approximately 50% of the heavy chains. The glycan was of the high-mannose type, with some galactose present, and the size ranged from Hex(6)GlcNAc(2) to Hex(15)GlcNAc(2). An aglycosyl mutant form of antibody was also produced. No significant difference between the glycosylated antibody produced by Aspergillus and that produced by mammalian cell cultures was observed in tests for affinity, avidity, pharmacokinetics, or antibody-dependent cellular cytotoxicity function. PMID- 15128506 TI - Impact of soil drying-rewetting stress on microbial communities and activities and on degradation of two crop protection products. AB - Prior to registration of crop protection products (CPPs) their persistence in soil has to be determined under defined conditions. For this purpose, soils are collected in the field and stored for up to 3 months prior to the tests. During storage, stresses like drying may induce changes in microbiological soil characteristics (MSCs) and thus may influence CPP degradation rates. We investigated the influence of soil storage-related stress on the resistance and resilience of different MSCs by assessing the impact of a single severe drying rewetting cycle and by monitoring recovery from this event for 34 days. The degradation and mineralization of the fungicide metalaxyl-M and the insecticide lufenuron were delayed by factors of 1.5 to 5.4 in the dried and rewetted soil compared to the degradation and mineralization in an undisturbed reference. The microbial biomass, as estimated by direct cell counting and from the soil DNA content, decreased on average by 51 and 24%, respectively. The bulk microbial activities, as determined by measuring substrate-induced respiration and fluorescein diacetate hydrolysis, increased after rewetting and recovered completely within 6 days after reequilibration. The effects on Bacteria, Archaea, and Pseudomonas were investigated by performing PCR amplification of 16S rRNA genes and reverse-transcribed 16S rRNA, followed by restriction fragment length polymorphism (RFLP) and terminal RFLP (T-RFLP) fingerprinting. Statistical analyses of RFLP and T-RFLP profiles indicated that specific groups in the microbial community were sensitive to the stress. In addition, evaluation of rRNA genes and rRNA as markers for monitoring the stress responses of microbial communities revealed overall similar sensitivities. We concluded that various structural and functional MSCs were not resistant to drying-rewetting stress and that resilience depended strongly on the parameter investigated. PMID- 15128507 TI - Cloning of the authentic bovine gene encoding pepsinogen a and its expression in microbial cells. AB - Bovine pepsin is the second major proteolytic activity of rennet obtained from young calves and is the main protease when it is extracted from adult animals, and it is well recognized that the proteolytic specificity of this enzyme improves the sensory properties of cheese during maturation. Pepsin is synthesized as an inactive precursor, pepsinogen, which is autocatalytically activated at the pH of calf abomasum. A cDNA coding for bovine pepsin was assembled by fusing the cDNA fragments from two different bovine expressed sequence tag libraries to synthetic DNA sequences based on the previously described N-terminal sequence of pepsinogen. The sequence of this cDNA clearly differs from the previously described partial bovine pepsinogen sequences, which actually are rabbit pepsinogen sequences. By cloning this cDNA in different vectors we produced functional bovine pepsinogen in Escherichia coli and Saccharomyces cerevisiae. The recombinant pepsinogen is activated by low pH, and the resulting mature pepsin has milk-clotting activity. Moreover, the mature enzyme generates digestion profiles with alpha-, beta-, or kappa-casein indistinguishable from those obtained with a natural pepsin preparation. The potential applications of this recombinant enzyme include cheese making and bioactive peptide production. One remarkable advantage of the recombinant enzyme for food applications is that there is no risk of transmission of bovine spongiform encephalopathy. PMID- 15128508 TI - A novel alpha-Proteobacterium resides in the mitochondria of ovarian cells of the tick Ixodes ricinus. AB - An intracellular bacterium from Ixodes ricinus ticks collected in Italy was characterized by electron microscopy (EM), PCR sequencing of the 16S rRNA gene, molecular phylogenetic analysis, and in situ hybridization (ISH). This bacterium was shown by EM to be present in the cytoplasm, as well as in the mitochondria of ovarian cells. When universal 16S rRNA bacterial primers were used, PCR amplification of ovarian DNA followed by cloning and sequencing resulted in the same sequence being found in each sample. Phylogenetic analysis of this sequence showed that the bacterium from which it was derived, tentatively designated IricES1, is part of a novel clade in the alpha subdivision of the Proteobacterium: ISH and PCR assays of various tissues performed with oligonucleotides specific for the IricES1 16S rRNA showed that IricES1 is restricted to ovarian cells. Based on the results obtained, we inferred that the bacteria seen by EM in ovarian cells are a single type of bacteria, corresponding to IricES1. PCR screening of 166 ticks from various parts of Italy and one site in England showed that IricES1 was present in 96% of adult females and 44% of nymphs (unsexed). No adult males were found to be infected. Despite the apparent parasitism of host mitochondria by IricES1, the available information suggests that the bacterium has an obligate relationship with its host, although this must be confirmed. PMID- 15128509 TI - Bacterial community dynamics and hydrocarbon degradation during a field-scale evaluation of bioremediation on a mudflat beach contaminated with buried oil. AB - A field-scale experiment with a complete randomized block design was performed to study the degradation of buried oil on a shoreline over a period of almost 1 year. The following four treatments were examined in three replicate blocks: two levels of fertilizer treatment of oil-treated plots, one receiving a weekly application of liquid fertilizer and the other treated with a slow-release fertilizer; and two controls, one not treated with oil and the other treated with oil but not with fertilizer. Oil degradation was monitored by measuring carbon dioxide evolution and by chemical analysis of the oil. Buried oil was degraded to a significantly greater extent in fertilized plots, but no differences in oil chemistry were observed between the two different fertilizer treatments, although carbon dioxide production was significantly higher in the oil-treated plots that were treated with slow-release fertilizer during the first 14 days of the experiment. Bacterial communities present in the beach sediments were profiled by denaturing gradient gel electrophoresis (DGGE) analysis of PCR-amplified 16S rRNA gene fragments and 16S rRNA amplified by reverse transcriptase PCR. Similarities between the DGGE profiles were calculated, and similarity matrices were subjected to statistical analysis. These analyses showed that although significant hydrocarbon degradation occurred both in plots treated with oil alone and in the plots treated with oil and liquid fertilizer, the bacterial community structure in these plots was, in general, not significantly different from that in the control plots that were not treated with oil and did not change over time. In contrast, the bacterial community structure in the plots treated with oil and slow-release fertilizer changed rapidly, and there were significant differences over time, as well as between blocks and even within plots. The differences were probably related to the higher concentrations of nutrients measured in interstitial water from the plots treated with slow-release fertilizer. Bacteria with 16S rRNA sequences closely related (>99.7% identity) to Alcanivorax borkumensis and Pseudomonas stutzeri sequences dominated during the initial phase of oil degradation in the plots treated with slow-release fertilizer. Field data were compared to the results of previous laboratory microcosm experiments, which revealed significant differences. PMID- 15128510 TI - Response of Archaeal communities in beach sediments to spilled oil and bioremediation. AB - While the contribution of Bacteria to bioremediation of oil-contaminated shorelines is well established, the response of Archaea to spilled oil and bioremediation treatments is unknown. The relationship between archaeal community structure and oil spill bioremediation was examined in laboratory microcosms and in a bioremediation field trial. 16S rRNA gene-based PCR and denaturing gradient gel analysis revealed that the archaeal community in oil-free laboratory microcosms was stable for 26 days. In contrast, in oil-polluted microcosms a dramatic decrease in the ability to detect Archaea was observed, and it was not possible to amplify fragments of archaeal 16S rRNA genes from samples taken from microcosms treated with oil. This was the case irrespective of whether a bioremediation treatment (addition of inorganic nutrients) was applied. Since rapid oil biodegradation occurred in nutrient-treated microcosms, we concluded that Archaea are unlikely to play a role in oil degradation in beach ecosystems. A clear-cut relationship between the presence of oil and the absence of Archaea was not apparent in the field experiment. This may have been related to continuous inoculation of beach sediments in the field with Archaea from seawater or invertebrates and shows that the reestablishment of Archaea following bioremediation cannot be used as a determinant of ecosystem recovery following bioremediation. Comparative 16S rRNA sequence analysis showed that the majority of the Archaea detected (94%) belonged to a novel, distinct cluster of group II uncultured Euryarchaeota, which exhibited less than 87% identity to previously described sequences. A minor contribution of group I uncultured Crenarchaeota was observed. PMID- 15128511 TI - Automated purification and suspension array detection of 16S rRNA from soil and sediment extracts by using tunable surface microparticles. AB - Autonomous, field-deployable molecular detection systems require seamless integration of complex biochemical solutions and physical or mechanical processing steps. In an attempt to simplify the fluidic requirements for integrated biodetection systems, we used tunable surface microparticles both as an rRNA affinity purification resin in a renewable microcolumn sample preparation system and as the sensor surface in a flow cytometer detector. The tunable surface detection limits in both low- and high-salt buffers were 1 ng of total RNA ( approximately 10(4) cell equivalents) in 15-min test tube hybridizations and 10 ng of total RNA ( approximately 10(5) cell equivalents) in hybridizations with the automated system (30-s contact time). RNA fragmentation was essential for achieving tunable surface suspension array specificity. Chaperone probes reduced but did not completely eliminate cross-hybridization, even with probes sharing <50% identity to target sequences. Nonpurified environmental extracts did not irreparably affect our ability to classify color-coded microparticles, but residual environmental constituents significantly quenched the Alexa-532 reporter fluor. Modulating surface charge did not influence the interaction of soluble environmental contaminants with conjugated beads. The automated system greatly reduced the effects of fluorescence quenching, especially in the soil background. The automated system was as efficacious as manual methods for simultaneous sample purification, hybridization, and washing prior to flow cytometry detection. The implications of unexpected target cross-hybridization and fluorescence quenching are discussed relative to the design and implementation of an integrated microbial monitoring system. PMID- 15128512 TI - Improved production of heterologous proteins by a glucose repression-defective mutant of Kluyveromyces lactis. AB - The secreted production of heterologous proteins in Kluyveromyces lactis was studied. A glucoamylase (GAA) from the yeast Arxula adeninivorans was used as a reporter protein for the study of the secretion efficiencies of several wild-type and mutant strains of K. lactis. The expression of the reporter protein was placed under the control of the strong promoter of the glyceraldehyde-3-phosphate dehydrogenase of Saccharomyces cerevisiae. Among the laboratory strains tested, strain JA6 was the best producer of GAA. Since this strain is known to be highly sensitive to glucose repression and since this is an undesired trait for biomass oriented applications, we examined heterologous protein production by using glucose repression-defective mutants isolated from this strain. One of them, a mutant carrying a dgr151-1 mutation, showed a significantly improved capability of producing heterologous proteins such as GAA, human serum albumin, and human interleukin-1beta compared to the parent strain. dgr151-1 is an allele of RAG5, the gene encoding the only hexokinase present in K. lactis (a homologue of S. cerevisiae HXK2). The mutation in this strain was mapped to nucleotide position +527, resulting in a change from glycine to aspartic acid within the highly conserved kinase domain. Cells carrying the dgr151-1 allele also showed a reduction in N- and O-glycosylation. Therefore, the dgr151 strain may be a promising host for the production of heterologous proteins, especially when the hyperglycosylation of recombinant proteins must be avoided. PMID- 15128513 TI - In vivo synthesis of mammalian-like, hybrid-type N-glycans in Pichia pastoris. AB - The Pichia pastoris N-glycosylation pathway is only partially homologous to the pathway in human cells. In the Golgi apparatus, human cells synthesize complex oligosaccharides, whereas Pichia cells form mannose structures that can contain up to 40 mannose residues. This hypermannosylation of secreted glycoproteins hampers the downstream processing of heterologously expressed glycoproteins and leads to the production of protein-based therapeutic agents that are rapidly cleared from the blood because of the presence of terminal mannose residues. Here, we describe engineering of the P. pastoris N-glycosylation pathway to produce nonhyperglycosylated hybrid glycans. This was accomplished by inactivation of OCH1 and overexpression of an alpha-1,2-mannosidase retained in the endoplasmic reticulum and N-acetylglucosaminyltransferase I and beta-1,4 galactosyltransferase retained in the Golgi apparatus. The engineered strain synthesized a nonsialylated hybrid-type N-linked oligosaccharide structure on its glycoproteins. The procedures which we developed allow glycan engineering of any P. pastoris expression strain and can yield up to 90% homogeneous protein-linked oligosaccharides. PMID- 15128514 TI - Structure-function analysis of immunity proteins of pediocin-like bacteriocins: C terminal parts of immunity proteins are involved in specific recognition of cognate bacteriocins. AB - The immunity proteins of pediocin-like bacteriocins show a high degree of specificity with respect to the pediocin-like bacteriocin they recognize and confer immunity to. The aim of this study was to identify regions of the immunity proteins that are involved in this specific recognition. Six different hybrid immunity proteins were constructed from three different pediocin-like bacteriocin immunity proteins that have similar sequences but confer resistance to different bacteriocins. These hybrid immunity proteins were then tested for their ability to confer immunity to various pediocin-like bacteriocins. The specificities of the hybrid immunity proteins proved to be similar to those of the immunity proteins from which the C-terminal halves were derived, thus revealing that the C terminal half of immunity proteins for pediocin-like bacteriocins contains a domain that is involved in specific recognition of the bacteriocins they confer immunity to. Moreover, the results also revealed that the effectiveness of an immunity protein is strain dependent and that its functionality thus depends in part on interplay with strain-dependent factors. To further investigate the structure-function relationship of these immunity proteins, the enterocin A and leucocin A immunity proteins (EntA-im and LeuA-im) were purified to homogeneity and structurally analyzed under various conditions by Circular dichroism (CD) spectroscopy. The results revealed that both immunity proteins are alpha-helical and well structured in an aqueous environment, the denaturing temperature being 78.5 degrees C for EntA-im and 58.0 degrees C for LeuA-im. The CD spectra also revealed that there was no further increase in the structuring or alpha-helical content when the immunity proteins were exposed to dodecylphosphocholine micelles or dioleoyl-L-alpha-phosphatidyl-DL-glycerol (DOPG) liposomes, indicating that the immunity proteins, in contrast to the bacteriocins, do not interact extensively with membranes. They may nevertheless be loosely associated with the membrane, possibly as peripheral membrane proteins, thus enabling them to interact with their cognate bacteriocin. PMID- 15128515 TI - Regulation of fumonisin biosynthesis in Fusarium verticillioides by a zinc binuclear cluster-type gene, ZFR1. AB - Fusarium verticillioides, a pathogen of maize, produces a class of mycotoxins called fumonisins in infected kernels. In this study, a candidate regulatory gene, ZFR1, was identified in an expressed sequence tag library enriched for transcripts expressed by F. verticillioides during fumonisin B(1) (FB(1)) biosynthesis. ZFR1 deletion mutants exhibited normal growth and development on maize kernels, but fumonisin production was reduced to less than 10% of that of the wild-type strain. ZFR1 encodes a putative protein of 705 amino acids with sequence similarity to the Zn(II)2Cys6 binuclear cluster family that are regulators of both primary and secondary metabolism in fungi. Expression of ZFR1 in colonized germ and degermed kernel tissues correlated with FB(1) levels. Overexpression of ZFR1 in zfr1 mutants restored FB(1) production to wild-type levels; however, FB(1) was not restored in an fcc1 (Fusarium C-type cyclin) mutant by overexpression of ZFR1. The results of this study indicate that ZFR1 is a positive regulator of FB(1) biosynthesis in F. verticillioides and suggest that FCC1 is required for ZFR1 function. PMID- 15128516 TI - Heat shock protein-mediated resistance to high hydrostatic pressure in Escherichia coli. AB - A random library of Escherichia coli MG1655 genomic fragments fused to a promoterless green fluorescent protein (GFP) gene was constructed and screened by differential fluorescence induction for promoters that are induced after exposure to a sublethal high hydrostatic pressure stress. This screening yielded three promoters of genes belonging to the heat shock regulon (dnaK, lon, clpPX), suggesting a role for heat shock proteins in protection against, and/or repair of, damage caused by high pressure. Several further observations provide additional support for this hypothesis: (i). the expression of rpoH, encoding the heat shock-specific sigma factor sigma(32), was also induced by high pressure; (ii). heat shock rendered E. coli significantly more resistant to subsequent high pressure inactivation, and this heat shock-induced pressure resistance followed the same time course as the induction of heat shock genes; (iii). basal expression levels of GFP from heat shock promoters, and expression of several heat shock proteins as determined by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis of proteins extracted from pulse-labeled cells, was increased in three previously isolated pressure-resistant mutants of E. coli compared to wild-type levels. PMID- 15128517 TI - Bacterial communities associated with flowering plants of the Ni hyperaccumulator Thlaspi goesingense. AB - Thlaspi goesingense is able to hyperaccumulate extremely high concentrations of Ni when grown in ultramafic soils. Recently it has been shown that rhizosphere bacteria may increase the heavy metal concentrations in hyperaccumulator plants significantly, whereas the role of endophytes has not been investigated yet. In this study the rhizosphere and shoot-associated (endophytic) bacteria colonizing T. goesingense were characterized in detail by using both cultivation and cultivation-independent techniques. Bacteria were identified by 16S rRNA sequence analysis, and isolates were further characterized regarding characteristics that may be relevant for a beneficial plant-microbe interaction-Ni tolerance, 1 aminocyclopropane-1-carboxylic acid (ACC) deaminase and siderophore production. In the rhizosphere a high percentage of bacteria belonging to the Holophaga/Acidobacterium division and alpha-Proteobacteria were found. In addition, high-G+C gram-positive bacteria, Verrucomicrobia, and microbes of the Cytophaga/Flexibacter/Bacteroides division colonized the rhizosphere. The community structure of shoot-associated bacteria was highly different. The majority of clones affiliated with the Proteobacteria, but also bacteria belonging to the Cytophaga/Flexibacter/Bacteroides division, the Holophaga/Acidobacterium division, and the low-G+C gram-positive bacteria, were frequently found. A high number of highly related Sphingomonas 16S rRNA gene sequences were detected, which were also obtained by the cultivation of endophytes. Rhizosphere isolates belonged mainly to the genera Methylobacterium, Rhodococcus, and Okibacterium, whereas the majority of endophytes showed high levels of similarity to Methylobacterium mesophilicum. Additionally, Sphingomonas spp. were abundant. Isolates were resistant to Ni concentrations between 5 and 12 mM; however, endophytes generally tolerated higher Ni levels than rhizosphere bacteria. Almost all bacteria were able to produce siderophores. Various strains, particularly endophytes, were able to grow on ACC as the sole nitrogen source. PMID- 15128518 TI - Detection of enteric viruses in shellfish from the Norwegian coast. AB - Common blue mussels (Mytilus edulis), horse mussels (Modiolus modiolus), and flat oysters (Ostrea edulis) obtained from various harvesting and commercial production sites along the Norwegian coast were screened for the presence of norovirus by a real-time reverse transcription (RT)-nested PCR assay and for possible indicators of fecal contamination, i.e., for F-specific RNA bacteriophages (F-RNA phages) by plaque assay and for human adenoviruses and human circoviruses by nested PCR assay. The aims were to obtain relevant information for assessing the risk of transmission of enteric viruses by shellfish and to investigate the potential of various indicator viruses in routine screening. Noroviruses were detected in 6.8% of the samples, and the indicators were detected in 23.8% (F-RNA phages), 18.6% (adenoviruses), and 8.0% (circoviruses) of the samples. A seasonal variation was observed, with the exception of circoviruses, with more positive samples in the winter. A positive correlation was found between F-RNA phages and noroviruses. However, F-RNA phages were present in only 43% of the norovirus-positive samples. The results show that mussels from the Norwegian coast can constitute a risk of infection with enteric viruses and that routine testing of samples may be justified. Advantages and disadvantages of various options for screening are discussed. PMID- 15128519 TI - Detection of enterotoxigenic Clostridium perfringens type A isolates in American retail foods. AB - Currently there is only limited understanding of the reservoirs for Clostridium perfringens type A food poisoning. A recent survey (Y.-T. Lin and R. Labbe, Appl. Environ. Microbiol. 69:1642-1646, 2003) of non-outbreak American retail foods did not identify the presence of a single C. perfringens isolate carrying the enterotoxin gene (cpe) necessary for causing food poisoning. The present study revisited this issue, using revised methodology and food sampling strategies. In our survey, cpe-positive C. perfringens isolates were detected in approximately 1.4% of approximately 900 surveyed non-outbreak American retail foods. Interestingly, those enterotoxigenic isolates in non-outbreak foods appear indistinguishable from C. perfringens isolates known to cause food poisoning outbreaks: i.e., the enterotoxigenic retail food isolates all carry a chromosomal cpe gene, are classified as type A, and exhibit exceptional heat resistance. Collectively, these findings indicate that some American foods are contaminated, at the time of retail purchase, with C. perfringens isolates having full potential to cause food poisoning. Furthermore, demonstrating that type A isolates carrying a chromosomal cpe gene are the enterotoxigenic isolates most commonly present in foods helps to explain why these isolates (rather than type A isolates carrying a plasmid cpe gene or cpe-positive type C or D isolates) are strongly associated with food poisoning outbreaks. Finally, since type A chromosomal cpe isolates present in the surveyed raw foods exhibited strong heat resistance, it appears that exceptional heat resistance is not a survivor trait selected for by cooking but is instead an intrinsic trait possessed by many type A chromosomal cpe isolates. PMID- 15128520 TI - Responses of active bacterial and fungal communities in soils under winter wheat to different fertilizer and pesticide regimens. AB - The composition of the active microbial (bacterial and fungal) soil community in an arable wheat field subjected to different management practices was examined at five times during a 1-year period. Field sections were fertilized either at good agricultural practice (GAP) levels or at reduced levels (0.5x GAP) and were inoculated with vesicular arbuscular mycorrhizae (VAM) at the same time. Field subsections were treated either with or without pesticides. Changes in the active microbial communities were investigated by denaturing gradient gel electrophoresis analysis of reverse transcription-PCR-amplified 16S and 18S rRNA. Microbial community structure was primarily determined by season, and the seasonal trends were similar for the fungal and bacterial components. Between sample microbial heterogeneity decreased under a mature crop in the summer but increased following harvesting and plowing. Although similar overall trends were seen for the two microbial components, sample variability was greater for the fungal community than for the bacterial community. The greatest management effects were due to GAP fertilization, which caused increases in the bacterial numbers in the total and culturable communities. Microbial biomass similarly increased. GAP fertilization also caused large shifts in both the active bacterial community structure and the active fungal community structure and additionally resulted in a decrease in the heterogeneity of the active bacterial community. Pesticide addition did not significantly affect bacterial numbers or heterogeneity, but it led to major shifts in the active soil bacterial community structure. PCR primers specific for Glomales 25S rRNA genes were used to monitor the VAM population following inoculation. Glomales were detected initially only in VAM-inoculated field sections but were subsequently detected in noninoculated field sections as the season progressed. After plowing, the level of Glomales was reduced in noninoculated field sections but remained high in VAM-inoculated field sections. Inoculation of VAM correlated with elevated soil phosphate and carbon levels. PMID- 15128522 TI - Modification of spatial distribution of 2,4-dichlorophenoxyacetic acid degrader microhabitats during growth in soil columns. AB - Bacterial processes in soil, including biodegradation, require contact between bacteria and substrates. Knowledge of the three-dimensional spatial distribution of bacteria at the microscale is necessary to understand and predict such processes. Using a soil microsampling strategy combined with a mathematical spatial analysis, we studied the spatial distribution of 2,4 dichlorophenoxyacetic acid (2,4-D) degrader microhabitats as a function of 2,4-D degrader abundance. Soil columns that allowed natural flow were percolated with 2,4-D to increase the 2,4-D degrader abundance. Hundreds of soil microsamples (minimum diameter, 125 microm) were collected and transferred to culture medium to check for the presence of 2,4-D degraders. Spatial distributions of bacterial microhabitats were characterized by determining the average size of colonized soil patches and the average number of patches per gram of soil. The spatial distribution of 2,4-D degrader microhabitats was not affected by water flow, but there was an overall increase in colonized patch sizes after 2,4-D amendment; colonized microsamples were dispersed in the soil at low 2,4-D degrader densities and clustered in patches that were more than 0.5 mm in diameter at higher densities. During growth, spreading of 2,4-D degraders within the soil and an increase in 2,4-D degradation were observed. We hypothesized that spreading of the bacteria increased the probability of encounters with 2,4-D and resulted in better interception of the degradable substrate. This work showed that characterization of bacterial microscale spatial distribution is relevant to microbial ecology studies. It improved quantitative bacterial microhabitat description and suggested that sporadic movement of cells occurs. Furthermore, it offered perspectives for linking microbial function to the soil physicochemical environment. PMID- 15128521 TI - Behavior of an Aeromonas hydrophila aroA live vaccine in water microcosms. AB - Genetically modified auxotrophic mutants of different fish pathogens have been used as live vaccines in laboratory experiments, but the behavior of the strains after release into aquatic ecosystems has not been characterized. We previously constructed and characterized an aroA mutant of Aeromonas hydrophila and studied the protection afforded by this mutant as a live vaccine in rainbow trout. In this work, we describe the survival of this strain in aquatic microcosms prepared from fish water tanks. The aroA mutant disappeared rapidly in nonfiltered, nonautoclaved fish tank water, declining below detection levels after 15 days, suggesting an inhibitory effect of the autochthonous microflora of the water. When the aroA strain was used to inoculate sterilized water, its culturability was lower than that of wild-type strain A. hydrophila AG2; after long periods of incubation, aroA cells were able to enter a viable but nonculturable state. Entry into this nonculturable state was accompanied by changes in the cell morphology from rods to spheres, but the cells appeared to remain potentially viable, as assessed by the preservation of cell membrane integrity. Supplementation of the culture medium with sodium pyruvate favored the culturability and resuscitation of the two A. hydrophila strains at low temperatures (6 and 16 degrees C). These results contribute to a better understanding of the behavior of the aroA strain in natural environments and suggest that the inactivation of the aroA gene may be beneficial for the safety of this live vaccine for aquacultures. PMID- 15128523 TI - Increased ATPase activity is responsible for acid sensitivity of nisin-resistant Listeria monocytogenes ATCC 700302. AB - The growth of the foodborne pathogen Listeria monocytogenes can be controlled by nisin, an antimicrobial peptide. A spontaneous mutant of L. monocytogenes shows both resistance to nisin and increased acid sensitivity compared to the wild type. Changes in the cell membrane correlated with nisin resistance, but the mechanism for acid sensitivity appears unrelated. When hydrochloric or lactic acid is added to cultures, intracellular ATP levels drop significantly in the mutant (P < 0.01) compared to the results seen with the wild type. Characterization of the F(0)F(1) ATPase, which hydrolyzes ATP to pump protons from the cell cytoplasm, shows that the enzyme is more active in the mutant than in the wild type. These data support a model in which the increased activity of the mutant ATPase upon acid addition depletes the cells' supply of ATP, resulting in cell death. PMID- 15128525 TI - Evaluation of a Clostridium perfringens predictive model, developed under isothermal conditions in broth, to predict growth in ground beef during cooling. AB - Proper temperature control is essential in minimizing Clostridium perfringens germination, growth, and toxin production. The U.S. Department of Agriculture (USDA) Food Safety and Inspection Service (FSIS) offers two options for the cooling of meat products: follow a standard time-temperature schedule or validate that alternative cooling regimens result in no more than a 1-log(10) CFU/g increase of C. perfringens and no growth of Clostridium botulinum. A mathematical model developed by Juneja et al. (Food Microbiol. 16:335-349, 1999) may be helpful in determining if the C. perfringens performance standard has been achieved, but this model has not been extensively validated. The objective of this study was to validate the Juneja 1999 model in ground beef under a variety of changing temperature and temperature abuse situations. The Juneja 1999 model consistently underpredicted growth of C. perfringens during exponential cooling of ground beef. The model also underpredicted growth of C. perfringens in ground beef cooled at two different rates. The results presented here show generally good agreement with published data on the growth of C. perfringens in similar products. The model error may be due to faster-than-expected exponential growth rates in ground beef during cooling or an error in the mathematical formulation of the model. PMID- 15128524 TI - Transformation of verapamil by Cunninghamella blakesleeana. AB - A filamentous fungus, Cunninghamella blakesleeana AS 3.153, was used as a microbial model of mammalian metabolism to transform verapamil, a calcium channel antagonist. The metabolites of verapamil were separated and assayed by the liquid chromatography-ion trap mass spectrometry method. After 96 h of incubation, nearly 93% of the original drug was metabolized to 23 metabolites. Five major metabolites were isolated by semipreparative high-performance liquid chromatography and were identified by proton nuclear magnetic resonance and electrospray mass spectrometry. Other metabolites were characterized according to their chromatographic behavior and mass spectral data. The major metabolic pathways of verapamil transformation by the fungus were N dealkylation, O demethylation, and sulfate conjugation. The phase I metabolites of verapamil (introduction of a functional group) by C. blakesleeana paralleled those in mammals; therefore, C. blakesleeana could be a useful tool for generating the mammalian phase I metabolites of verapamil. PMID- 15128526 TI - Lack of protective osmolytes limits final cell density and volumetric productivity of ethanologenic Escherichia coli KO11 during xylose fermentation. AB - Limited cell growth and the resulting low volumetric productivity of ethanologenic Escherichia coli KO11 in mineral salts medium containing xylose have been attributed to inadequate partitioning of carbon skeletons into the synthesis of glutamate and other products derived from the citrate arm of the anaerobic tricarboxylic acid pathway. The results of nuclear magnetic resonance investigations of intracellular osmolytes under different growth conditions coupled with those of studies using genetically modified strains have confirmed and extended this hypothesis. During anaerobic growth in mineral salts medium containing 9% xylose (600 mM) and 1% corn steep liquor, proline was the only abundant osmolyte (71.9 nmol x ml(-1) optical density at 550 nm [OD(550)] unit( 1)), and growth was limited. Under aerobic conditions in the same medium, twice the cell mass was produced, and cells contained a mixture of osmolytes: glutamate (17.0 nmol x ml(-1) OD(550) unit(-1)), trehalose (9.9 nmol x ml(-1) OD(550) unit( 1)), and betaine (19.8 nmol x ml(-1) OD(550) unit(-1)). Two independent genetic modifications of E. coli KO11 (functional expression of Bacillus subtilis citZ encoding NADH-insensitive citrate synthase; deletion of ackA encoding acetate kinase) and the addition of a metabolite, such as glutamate (11 mM) or acetate (24 mM), as a supplement each increased the intracellular glutamate pool during fermentation, doubled cell growth, and increased volumetric productivity. This apparent requirement for a larger glutamate pool for increased growth and volumetric productivity was completely eliminated by the addition of a protective osmolyte (2 mM betaine or 0.25 mM dimethylsulfoniopropionate), consistent with adaptation to osmotic stress rather than relief of a specific biosynthetic requirement. PMID- 15128527 TI - Dissimilatory arsenate reduction with sulfide as electron donor: experiments with mono lake water and Isolation of strain MLMS-1, a chemoautotrophic arsenate respirer. AB - Anoxic bottom water from Mono Lake, California, can biologically reduce added arsenate without any addition of electron donors. Of the possible in situ inorganic electron donors present, only sulfide was sufficiently abundant to drive this reaction. We tested the ability of sulfide to serve as an electron donor for arsenate reduction in experiments with lake water. Reduction of arsenate to arsenite occurred simultaneously with the removal of sulfide. No loss of sulfide occurred in controls without arsenate or in sterilized samples containing both arsenate and sulfide. The rate of arsenate reduction in lake water was dependent on the amount of available arsenate. We enriched for a bacterium that could achieve growth with sulfide and arsenate in a defined, mineral medium and purified it by serial dilution. The isolate, strain MLMS-1, is a gram-negative, motile curved rod that grows by oxidizing sulfide to sulfate while reducing arsenate to arsenite. Chemoautotrophy was confirmed by the incorporation of H(14)CO(3)(-) into dark-incubated cells, but preliminary gene probing tests with primers for ribulose-1,5-biphosphate carboxylase/oxygenase did not yield PCR-amplified products. Alignment of 16S rRNA sequences indicated that strain MLMS-1 was in the delta-Proteobacteria, located near sulfate reducers like Desulfobulbus sp. (88 to 90% similarity) but more closely related (97%) to unidentified sequences amplified previously from Mono Lake. However, strain MLMS 1 does not grow with sulfate as its electron acceptor. PMID- 15128528 TI - Effect of adaptation to ethanol on cytoplasmic and membrane protein profiles of Oenococcus oeni. AB - The practical application of commercial malolactic starter cultures of Oenococcus oeni surviving direct inoculation in wine requires insight into mechanisms of ethanol toxicity and of acquired ethanol tolerance in this organism. Therefore, the site-specific location of proteins involved in ethanol adaptation, including cytoplasmic, membrane-associated, and integral membrane proteins, was investigated. Ethanol triggers alterations in protein patterns of O. oeni cells stressed with 12% ethanol for 1 h and those of cells grown in the presence of 8% ethanol. Levels of inosine-5'-monophosphate dehydrogenase and phosphogluconate dehydrogenase, which generate reduced nicotinamide nucleotides, were decreased during growth in the presence of ethanol, while glutathione reductase, which consumes NADPH, was induced, suggesting that maintenance of the redox balance plays an important role in ethanol adaptation. Phosphoenolpyruvate:mannose phosphotransferase system (PTS) components of mannose PTS, including the phosphocarrier protein HPr and EII(Man), were lacking in ethanol-adapted cells, providing strong evidence that mannose PTS is absent in ethanol-adapted cells, and this represents a metabolic advantage to O. oeni cells during malolactic fermentation. In cells grown in the presence of ethanol, a large increase in the number of membrane-associated proteins was observed. Interestingly, two of these proteins, dTDT-glucose-4,6-dehydratase and D-alanine:D-alanine ligase, are known to be involved in cell wall biosynthesis. Using a proteomic approach, we provide evidence for an active ethanol adaptation response of O. oeni at the cytoplasmic and membrane protein levels. PMID- 15128529 TI - Subpopulation characteristics of egg-contaminating Salmonella enterica serovar Enteritidis as defined by the lipopolysaccharide O chain. AB - Characterization of Salmonella enterica serovar Enteritidis was refined by incorporating new data from isolates obtained from avian sources, from the spleens of naturally infected mice, and from the United Kingdom into an existing lipopolysaccharide (LPS) O-chain compositional database. From least to greatest, the probability of avian isolates producing high-molecular-mass LPS O chain ranked as follows: pooled kidney, liver, and spleen; intestine; cecum; ovary and oviduct; albumen; yolk; and whole egg. Mouse isolates were most like avian intestinal samples, whereas United Kingdom isolates were most like those from the avian reproductive tract and egg. Non-reproductive tract organ isolates had significant loss of O chain. Isogenic isolates that varied in ability to make biofilm and to be orally invasive produced different O-chain structures at 25 degrees C but not at 37 degrees C. Hens infected at a 91:9 biofilm-positive/ negative colony phenotype ratio yielded only the negative phenotype from eggs. These results indicate that the environment within the hen applies stringent selection pressure on subpopulations of S. enterica serovar Enteritidis at certain points in the infection pathway that ends in egg contamination. The avian cecum, rather than the intestines, is the early interface between the environment and the host that supports emergence of subpopulation diversity. These results suggest that diet and other factors that alter cecal physiology should be investigated as a means to reduce egg contamination. PMID- 15128530 TI - Cloning, overexpression, and characterization of a novel thermostable penicillin G acylase from Achromobacter xylosoxidans: probing the molecular basis for its high thermostability. AB - The gene encoding a novel penicillin G acylase (PGA), designated pgaW, was cloned from Achromobacter xylosoxidans and overexpressed in Escherichia coli. The pgaW gene contains an open reading frame of 2586 nucleotides. The deduced protein sequence encoded by pgaW has about 50% amino acid identity to several well characterized PGAs, including those of Providencia rettgeri, Kluyvera cryocrescens, and Escherichia coli. Biochemical studies showed that the optimal temperature for this novel PGA (PGA650) activity is greater than 60 degrees C and its half-life of inactivation at 55 degrees C is four times longer than that of another previously reported thermostable PGA from Alcaligenes faecalis (R. M. D. Verhaert, A. M. Riemens, J. V. R. Laan, J. V. Duin, and W. J. Quax, Appl. Environ. Microbiol. 63:3412-3418, 1997). To our knowledge, this is the most thermostable PGA ever characterized. To explore the molecular basis of the higher thermostability of PGA650, homology structural modeling and amino acid composition analyses were performed. The results suggested that the increased number of buried ion pair networks, lower N and Q contents, excessive arginine residues, and remarkably high content of proline residues in the structure of PGA650 could contribute to its high thermostability. The unique characteristic of higher thermostability of this novel PGA provides some advantages for its potential application in industry. PMID- 15128531 TI - Two distinct pathways for trehalose assimilation in the yeast Saccharomyces cerevisiae. AB - The yeast Saccharomyces cerevisiae can synthesize trehalose and also use this disaccharide as a carbon source for growth. However, the molecular mechanism by which extracellular trehalose can be transported to the vacuole and degraded by the acid trehalase Ath1p is not clear. By using an adaptation of the assay of invertase on whole cells with NaF, we showed that more than 90% of the activity of Ath1p is extracellular, splitting of the disaccharide into glucose. We also found that Agt1p-mediated trehalose transport and the hydrolysis of the disaccharide by the cytosolic neutral trehalase Nth1p are coupled and represent a second, independent pathway, although there are several constraints on this alternative route. First, the AGT1/MAL11 gene is controlled by the MAL system, and Agt1p was active in neither non-maltose-fermenting nor maltose-inducible strains. Second, Agt1p rapidly lost activity during growth on trehalose, by a mechanism similar to the sugar-induced inactivation of the maltose permease. Finally, both pathways are highly pH sensitive and effective growth on trehalose occurred only when the medium was buffered at around pH 5.0. The catabolism of trehalose was purely oxidative, and since levels of Ath1p limit the glucose flux in the cells, batch cultures on trehalose may provide a useful alternative to glucose-limited chemostat cultures for investigation of metabolic responses in yeast. PMID- 15128532 TI - Acquisition of an Agrobacterium Ri plasmid and pathogenicity by other alpha Proteobacteria in cucumber and tomato crops affected by root mat. AB - Root mat of cucumbers and tomatoes has previously been shown to be caused by Agrobacterium radiobacter strains harboring a root-inducing Ri plasmid (pRi). Nine other pRi-harboring alpha-Proteobacteria have subsequently been isolated from root mat-infected crops. Fatty acid profiling and partial 16S rRNA sequence analysis identified three of these strains as being in the genus Ochrobactrum, five as being in the genus Rhizobium, and one as being in the genus Sinorhizobium: An in vitro pathogenicity test involving inoculation of cucumber cotyledons was developed. All pRi-harboring alpha-Proteobacteria induced typical root mat symptoms from the cotyledons. Average transformation rates for rhizogenic Ochrobactrum (46%) and Rhizobium (44%) strains were lower than those observed for rhizogenic A. radiobacter strains (64%). However, individual strains from these three genera all had transformation rates comparable to those observed from cotyledons inoculated with a rhizogenic Sinorhizobium strain (75%). PMID- 15128533 TI - Detection of molecular diversity in Bacillus atrophaeus by amplified fragment length polymorphism analysis. AB - Phenotypically, Bacillus atrophaeus is indistinguishable from the type strain of Bacillus subtilis except by virtue of pigment production on certain media. Several pigmented variants of B. subtilis have been reclassified as B. atrophaeus, but several remain ambiguous in regard to their taxonomic placement. In this study, we examined strains within the American Type Culture Collection originally deposited as Bacillus globigii, B. subtilis var. niger, or Bacillus niger using 16S rRNA gene sequencing and amplified fragment length polymorphism (AFLP) analysis to determine the level of molecular diversity among these strains and their relationship with closely related taxa. The 16S rRNA gene sequences revealed little variation with one base substitution between the B. atrophaeus type strain ATCC 49337 and the other pigmented bacilli. AFLP analysis produced high-quality DNA fingerprints with sufficient polymorphism to reveal strain-level variation. Cluster analysis of Dice similarity coefficients revealed that three strains, ATCC 31028, ATCC 49760, and ATCC 49822, are much more closely related to B. atrophaeus than to B. subtilis and should be reclassified as B. atrophaeus. A very closely related cluster of B. atrophaeus strains was also observed; this cluster was genetically distinct from the type strain. The level of variation between the two groups was approximately the same as the level of variation observed between members of the two B. subtilis subspecies, subtilis and spizizenii. It is proposed that the cluster of strains typified by ATCC 9372 be designated a new subspecies, B. atrophaeus subsp. globigii. PMID- 15128534 TI - Spatial distribution and stability of the eight microbial species of the altered schaedler flora in the mouse gastrointestinal tract. AB - The overall complexity of the microbial communities in the gastrointestinal (GI) tracts of mammals has hindered observations of dynamics and interactions of individual bacterial populations. However, such information is crucial for understanding the diverse disease-causing and protective roles that gut microbiota play in their hosts. Here, we determine the spatial distribution, interanimal variation, and persistence of bacteria in the most complex defined flora (gnotobiotic) model system to date, viz., mice colonized with the eight strains of the altered Schaedler flora (ASF). Quantitative PCR protocols based on the 16S rRNA sequence of each ASF strain were developed and optimized to specifically detect as few as 10 copies of each target. Total numbers of the ASF strains were determined in the different regions of the GI tracts of three C.B-17 SCID mice. Individual strain abundance was dependent on oxygen sensitivity, with microaerotolerant Lactobacillus murinus ASF361 present at 10(5) to 10(7) cells/g of tissue in the upper GI tract and obligate anaerobic ASF strains being predominant in the cecal and colonic flora at 10(8) to 10(10) cells/g of tissue. The variation between the three mice was small for most ASF strains, except for Clostridium sp. strain ASF502 and Bacteroides sp. strain ASF519 in the cecum. A comparison of the relative distribution of the ASF strains in feces and the colon indicated large differences, suggesting that fecal bacterial levels may provide a poor approximation of colonic bacterial levels. All ASF strains were detected by PCR in the feces of C57BL/6 restricted flora mice, which had been maintained in an isolator without sterile food, water, or bedding for several generations, providing evidence for the stability of these strains in the face of potential competition by bacteria introduced into the gut. PMID- 15128535 TI - Double-layer plaque assay for quantification of enteroviruses. AB - We describe here a double-layer plaque assay for the quantification of enteroviruses, combining a monolayer plaque assay and a suspended-cell plaque assay. The double-layer assay provides significantly greater counts than other methods of virus quantification of both suspensions of pure culture viruses and naturally occurring viruses. The counts obtained by this method are approximately one order of magnitude greater than those obtained with the more commonly used method, the monolayer plaque assay. We conclude that the methods available for quantifying viruses rank in efficiency as follows: double-layer plaque assay >or=suspended-cell plaque assay > counting cytopathogenic virus adsorbed to cellulose nitrate membrane filters >or= most probable number of cytopathogenic units > monolayer plaque assay. Moreover, the double-layer plaque assay allows the use of two different cell lines in the two layers. Using the human colonic carcinoma cell line CaCo2 facilitates the recovery of a greater number and diversity of naturally occurring enteroviruses in water than the monolayer agar method. In addition, the pretreatment of cells with 5-iodo-2'-deoxyuridine (IDU) prior to the quantification of enteroviruses by the double-layer plaque assay provides significantly higher recoveries than the use of IDU does with the other methods of quantification. PMID- 15128536 TI - New recombination methods for Sinorhizobium meliloti genetics. AB - The availability of bacterial genome sequences has created a need for improved methods for sequence-based functional analysis to facilitate moving from annotated DNA sequence to genetic materials for analyzing the roles that postulated genes play in bacterial phenotypes. A powerful cloning method that uses lambda integrase recombination to clone and manipulate DNA sequences has been adapted for use with the gram-negative alpha-proteobacterium Sinorhizobium meliloti in two ways that increase the utility of the system. Adding plasmid oriT sequences to a set of vehicles allows the plasmids to be transferred to S. meliloti by conjugation and also allows cloned genes to be recombined from one plasmid to another in vivo by a pentaparental mating protocol, saving considerable time and expense. In addition, vehicles that contain yeast Flp recombinase target recombination sequences allow the construction of deletion mutations where the end points of the deletions are located at the ends of the cloned genes. Several deletions were constructed in a cluster of 60 genes on the symbiotic plasmid (pSymA) of S. meliloti, predicted to code for a denitrification pathway. The mutations do not affect the ability of the bacteria to form nitrogen fixing nodules on Medicago sativa (alfalfa) roots. PMID- 15128537 TI - Transposon mutagenesis of the obligate intracellular pathogen Rickettsia prowazekii. AB - Genetic analysis of Rickettsia prowazekii has been hindered by the lack of selectable markers and efficient mechanisms for generating rickettsial gene knockouts. We have addressed these problems by adapting a gene that codes for rifampin resistance for expression in R. prowazekii and by incorporating this selection into a transposon mutagenesis system suitable for generating rickettsial gene knockouts. The arr-2 gene codes for an enzyme that ADP ribosylates rifampin, thereby destroying its antibacterial activity. Based on the published sequence, this gene was synthesized by PCR with overlapping primers that contained rickettsial codon usage base changes. This R. prowazekii-adapted arr-2 gene (Rparr-2) was placed downstream of the strong rickettsial rpsL promoter (rpsL(P)), and the entire construct was inserted into the Epicentre EZ::TN transposome system. A purified transposon containing rpsL(P)-Rparr-2 was combined with transposase, and the resulting DNA-protein complex (transposome) was electroporated into competent rickettsiae. Following selection with rifampin, rickettsiae with transposon insertions in the genome were identified by PCR and Southern blotting and the insertion sites were determined by rescue cloning and inverse PCR. Multiple insertions into widely spaced areas of the R. prowazekii genome were identified. Three insertions were identified within gene coding sequences. Transposomes provide a mechanism for generating random insertional mutations in R. prowazekii, thereby identifying nonessential rickettsial genes. PMID- 15128538 TI - Germination of penicillium paneum Conidia is regulated by 1-octen-3-ol, a volatile self-inhibitor. AB - Penicillium paneum is an important contaminant of cereal grains which is able to grow at low temperature, low pH, high levels of carbon dioxide, and under acid conditions. P. paneum produces mycotoxins, which may be harmful to animals and humans. We found that conidia in dense suspensions showed poor germination, suggesting the presence of a self-inhibitor. A volatile compound(s) produced by these high-density conditions also inhibited mycelial growth of different species of fungi belonging to a variety of genera, suggesting a broad action range. The heat-stable compound was isolated by successive centrifugation of the supernatant obtained from spore suspensions with a density of 10(9) conidia ml(-1). By using static headspace analyses, two major peaks were distinguished, with the highest production of these metabolites after 22 h of incubation at 25 degrees C and shaking at 140 rpm. Gas chromatography coupled with mass spectra analysis revealed the compounds to be 3-octanone and 1-octen-3-ol. Notably, only the latter compound appeared to block the germination process at different developmental stages of the conidia (swelling and germ tube formation). In this study, 1-octen-3-ol influenced different developmental processes during the P. paneum life cycle, including induction of microcycle conidiation and inhibition of spore germination. Therefore, the compound can be considered a fungal hormone during fungal development. PMID- 15128539 TI - Addition of aromatic substrates restores trichloroethylene degradation activity in Pseudomonas putida F1. AB - The rate of trichloroethylene (TCE) degradation by toluene dioxygenase (TDO) in resting cells of Pseudomonas putida F1 gradually decreased and eventually stopped within 1.5 h, as in previous reports. However, the subsequent addition of toluene, which is the principal substrate of TDO, resulted in its immediate degradation without a lag phase. After the consumption of toluene, degradation of TCE restarted at a rate similar to its initial degradation, suggesting that this degradation was mediated by TDO molecules that were present before the cessation of TCE degradation. The addition of benzene and cumene, which are also substrates of TDO, also caused restoration of TCE degradation activity: TCE was degraded simultaneously with cumene, and a larger amount of TCE was degraded after cumene was added than after toluene or benzene was added. But substrates that were expected to supply the cells with NADH or energy did not restore TCE degradation activity. This cycle of pseudoinactivation and restoration of TCE degradation was observed repeatedly without a significant decrease in the number of viable cells, even after six additions of toluene spread over 30 h. The results obtained in this study demonstrate a new type of restoration of TCE degradation that has not been previously reported. PMID- 15128541 TI - Influence of precipitation and soil on transport of fecal enterococci in fractured limestone aquifers. AB - Limestone aquifers provide the main drinking water resources of southern Italy. The groundwater is often contaminated by fecal bacteria because of the interaction between rocks having high permeability and microbial pollutants introduced into the environment by grazing and/or manure spreading. The microbial contamination of springwater in picnic areas located in high mountains can cause gastrointestinal illness. This study was carried out in order to analyze the interaction between Enterococcus faecalis and the soil of a limestone aquifer and to verify the influence of this interaction on the time dependence of groundwater contamination. E. faecalis was chosen because, in the study area involved, it represents a better indicator than Escherichia coli. The research was carried out through field (springwater monitoring) and laboratory experiments (column tests with intact soil blocks). The transport of bacterial cells through soil samples was analyzed by simulating an infiltration event that was monitored in the study area. Comparison of laboratory results with data acquired in the field showed that discontinuous precipitation caused an intermittent migration of microorganisms through the soil and produced, together with dispersion in the fractured medium (unsaturated and saturated zones), an articulated breakthrough at the spring. The short distances of bacterial transport in the study area produced a significant daily variability of bacterial contamination at the field scale. PMID- 15128540 TI - Prevalence of the Chloroflexi-related SAR202 bacterioplankton cluster throughout the mesopelagic zone and deep ocean. AB - Since their initial discovery in samples from the north Atlantic Ocean, 16S rRNA genes related to the environmental gene clone cluster known as SAR202 have been recovered from pelagic freshwater, marine sediment, soil, and deep subsurface terrestrial environments. Together, these clones form a major, monophyletic subgroup of the phylum Chloroflexi: While members of this diverse group are consistently identified in the marine environment, there are currently no cultured representatives, and very little is known about their distribution or abundance in the world's oceans. In this study, published and newly identified SAR202-related 16S rRNA gene sequences were used to further resolve the phylogeny of this cluster and to design taxon-specific oligonucleotide probes for fluorescence in situ hybridization. Direct cell counts from the Bermuda Atlantic time series study site in the north Atlantic Ocean, the Hawaii ocean time series site in the central Pacific Ocean, and along the Newport hydroline in eastern Pacific coastal waters showed that SAR202 cluster cells were most abundant below the deep chlorophyll maximum and that they persisted to 3600 m in the Atlantic Ocean and to 4000 m in the Pacific Ocean, the deepest samples used in this study. On average, members of the SAR202 group accounted for 10.2% (+/-5.7%) of all DNA containing bacterioplankton between 500 and 4000 m. PMID- 15128542 TI - Use of aqueous silver to enhance inactivation of coliphage MS-2 by UV disinfection. AB - A synergistic effect between silver and UV radiation has been observed that can appreciably enhance the effectiveness of UV radiation for inactivation of viruses. At a fluence of ca. 40 mJ/cm(2), the synergistic effect between silver and UV was observed at silver concentrations as low as 10 microg/liter (P < 0.0615). At the same fluence, an MS-2 inactivation of ca. 3.5 logs (99.97%) was achieved at a silver concentration of 0.1 mg/liter, a significant improvement (P < 0.0001) over the ca. 1.8-log (98.42%) inactivation of MS-2 at ca. 40 mJ/cm(2) in the absence of silver. Modified Chick-Watson kinetics were used to model the synergistic effect of silver and UV radiation. For an MS-2 inactivation of 4 logs (99.99%), the coefficient of dilution (n) was determined to be 0.31, which suggests that changes in fluence have a greater influence on inactivation than does a proportionate change in silver concentration. PMID- 15128543 TI - Nitrite elimination and hydrolytic ring cleavage in 2,4,6-trinitrophenol (picric acid) degradation. AB - Two hydrogenation reactions in the initial steps of degradation of 2,4,6 trinitrophenol produce the dihydride Meisenheimer complex of 2,4,6 trinitrophenol. The npdH gene (contained in the npd gene cluster of the 2,4,6 trinitrophenol-degrading strain Rhodococcus opacus HL PM-1) was shown here to encode a tautomerase, catalyzing a proton shift between the aci-nitro and the nitro forms of the dihydride Meisenheimer complex of 2,4,6-trinitrophenol. An enzyme (which eliminated nitrite from the aci-nitro form but not the nitro form of the dihydride complex of 2,4,6-trinitrophenol) was purified from the 2,4,6 trinitrophenol-degrading strain Nocardioides simplex FJ2-1A. The product of nitrite release was the hydride Meisenheimer complex of 2,4-dinitrophenol, which was hydrogenated to the dihydride Meisenheimer complex of 2,4-dinitrophenol by the hydride transferase I and the NADPH-dependent F(420) reductase from strain HL PM-1. At pH 7.5, the dihydride complex of 2,4-dinitrophenol is protonated to 2,4 dinitrocyclohexanone. A hydrolase was purified from strain FJ2-1A and shown to cleave 2,4-dinitrocyclohexanone hydrolytically to 4,6-dinitrohexanoate. PMID- 15128544 TI - Heterologous expression of lactose- and galactose-utilizing pathways from lactic acid bacteria in Corynebacterium glutamicum for production of lysine in whey. AB - The genetic determinants for lactose utilization from Lactobacillus delbrueckii subsp. bulgaricus ATCC 11842 and galactose utilization from Lactococcus lactis subsp. cremoris MG 1363 were heterologously expressed in the lysine-overproducing strain Corynebacterium glutamicum ATCC 21253. The C. glutamicum strains expressing the lactose permease and beta-galactosidase genes of L. delbrueckii subsp. bulgaricus exhibited beta-galactosidase activity in excess of 1000 Miller units/ml of cells and were able to grow in medium in which lactose was the sole carbon source. Similarly, C. glutamicum strains containing the lactococcal aldose 1-epimerase, galactokinase, UDP-glucose-1-P-uridylyltransferase, and UDP galactose-4-epimerase genes in association with the lactose permease and beta galactosidase genes exhibited beta-galactosidase levels in excess of 730 Miller units/ml of cells and were able to grow in medium in which galactose was the sole carbon source. When grown in whey-based medium, the engineered C. glutamicum strain produced lysine at concentrations of up to 2 mg/ml, which represented a 10 fold increase over the results obtained with the lactose- and galactose-negative control, C. glutamicum 21253. Despite their increased catabolic flexibility, however, the modified corynebacteria exhibited slower growth rates and plasmid instability. PMID- 15128546 TI - Assembly of G protein-coupled receptors onto nanosized bacterial magnetic particles using Mms16 as an anchor molecule. AB - G protein-coupled receptors (GPCRs) play a central role in a wide range of biological processes and are prime targets for drug discovery. GPCRs have large hydrophobic domains, and therefore purification of GPCRs from cells is frequently time-consuming and typically results in loss of native conformation. In this work, GPCRs have been successfully assembled into the lipid membrane of nanosized bacterial magnetic particles (BMPs) produced by the magnetic bacterium Magnetospirillum magneticum AMB-1. A BMP-specific protein, Mms16, was used as an anchor molecule, and localization of heterologous Mms16 on BMPs was confirmed by luciferase fusion studies. Stable luminescence was obtained from BMPs bearing Mms16 fused with luciferase at the C-terminal region. D1 dopamine receptor (D1R), a GPCR, was also efficiently assembled onto BMPs by using Mms16 as an anchor molecule. D1R-BMP complexes were simply extracted by magnetic separation from ruptured AMB-1 transformants. After washing, the complexes were ready to use for analysis. This system conveniently refines the native conformation of GPCRs without the need for detergent solubilization, purification, and reconstitution after cell disruption. PMID- 15128545 TI - Seasonal changes in an alpine soil bacterial community in the colorado rocky mountains. AB - The period when the snowpack melts in late spring is a dynamic time for alpine ecosystems. The large winter microbial community begins to turn over rapidly, releasing nutrients to plants. Past studies have shown that the soil microbial community in alpine dry meadows of the Colorado Rocky Mountains changes in biomass, function, broad-level structure, and fungal diversity between winter and early summer. However, little specific information exists on the diversity of the alpine bacterial community or how it changes during this ecologically important period. We constructed clone libraries of 16S ribosomal DNA from alpine soil collected in winter, spring, and summer. We also cultivated bacteria from the alpine soil and measured the seasonal abundance of selected cultured isolates in hybridization experiments. The uncultured bacterial communities changed between seasons in diversity and abundance within taxa. The Acidobacterium division was most abundant in the spring. The winter community had the highest proportion of Actinobacteria and members of the Cytophaga/Flexibacter/Bacteroides (CFB) division. The summer community had the highest proportion of the Verrucomicrobium division and of beta-PROTEOBACTERIA: As a whole, alpha-Proteobacteria were equally abundant in all seasons, although seasonal changes may have occurred within this group. A number of sequences from currently uncultivated divisions were found, including two novel candidate divisions. The cultured isolates belonged to the alpha-, beta-, and gamma-Proteobacteria, the Actinobacteria, and the CFB groups. The only uncultured sequences that were closely related to the isolates were from winter and spring libraries. Hybridization experiments showed that actinobacterial and beta-proteobacterial isolates were most abundant during winter, while the alpha- and gamma-proteobacterial isolates tested did not vary significantly. While the cultures and clone libraries produced generally distinct groups of organisms, the two approaches gave consistent accounts of seasonal changes in microbial diversity. PMID- 15128547 TI - New method of denitrification analysis of bradyrhizobium field isolates by gas chromatographic determination of (15)N-labeled N(2). AB - To evaluate the denitrification abilities of many Bradyrhizobium field isolates, we developed a new (15)N-labeled N(2) detection methodology, which is free from interference from atmospheric N(2) contamination. (30)N(2) ((15)N(15)N) and (29)N(2) ((15)N(14)N) were detected as an apparent peak by a gas chromatograph equipped with a thermal conductivity detector with N(2) gas having natural abundance of (15)N (0.366 atom%) as a carrier gas. The detection limit was 0.04% (30)N(2), and the linearity extended at least to 40% (30)N(2). When Bradyrhizobium japonicum USDA110 was grown in cultures anaerobically with (15)NO(3)(-), denitrification product ((30)N(2)) was detected stoichiometrically. A total of 65 isolates of soybean bradyrhizobia from two field sites in Japan were assayed by this method. The denitrification abilities were partly correlated with filed sites, Bradyrhizobium species, and the hup genotype. PMID- 15128548 TI - Metabolic engineering of a phosphoketolase pathway for pentose catabolism in Saccharomyces cerevisiae. AB - Low ethanol yields on xylose hamper economically viable ethanol production from hemicellulose-rich plant material with Saccharomyces cerevisiae. A major obstacle is the limited capacity of yeast for anaerobic reoxidation of NADH. Net reoxidation of NADH could potentially be achieved by channeling carbon fluxes through a recombinant phosphoketolase pathway. By heterologous expression of phosphotransacetylase and acetaldehyde dehydrogenase in combination with the native phosphoketolase, we installed a functional phosphoketolase pathway in the xylose-fermenting Saccharomyces cerevisiae strain TMB3001c. Consequently the ethanol yield was increased by 25% because less of the by-product xylitol was formed. The flux through the recombinant phosphoketolase pathway was about 30% of the optimum flux that would be required to completely eliminate xylitol and glycerol accumulation. Further overexpression of phosphoketolase, however, increased acetate accumulation and reduced the fermentation rate. By combining the phosphoketolase pathway with the ald6 mutation, which reduced acetate formation, a strain with an ethanol yield 20% higher and a xylose fermentation rate 40% higher than those of its parent was engineered. PMID- 15128549 TI - Homofermentative lactate production cannot sustain anaerobic growth of engineered Saccharomyces cerevisiae: possible consequence of energy-dependent lactate export. AB - Due to a growing market for the biodegradable and renewable polymer polylactic acid, the world demand for lactic acid is rapidly increasing. The tolerance of yeasts to low pH can benefit the process economy of lactic acid production by minimizing the need for neutralizing agents. Saccharomyces cerevisiae (CEN.PK background) was engineered to a homofermentative lactate-producing yeast via deletion of the three genes encoding pyruvate decarboxylase and the introduction of a heterologous lactate dehydrogenase (EC 1.1.1.27). Like all pyruvate decarboxylase-negative S. cerevisiae strains, the engineered strain required small amounts of acetate for the synthesis of cytosolic acetyl-coenzyme A. Exposure of aerobic glucose-limited chemostat cultures to excess glucose resulted in the immediate appearance of lactate as the major fermentation product. Ethanol formation was absent. However, the engineered strain could not grow anaerobically, and lactate production was strongly stimulated by oxygen. In addition, under all conditions examined, lactate production by the engineered strain was slower than alcoholic fermentation by the wild type. Despite the equivalence of alcoholic fermentation and lactate fermentation with respect to redox balance and ATP generation, studies on oxygen-limited chemostat cultures showed that lactate production does not contribute to the ATP economy of the engineered yeast. This absence of net ATP production is probably due to a metabolic energy requirement (directly or indirectly in the form of ATP) for lactate export. PMID- 15128550 TI - Structural and functional differences in two cyclic bacteriocins with the same sequences produced by lactobacilli. AB - Lactobacillus gasseri LA39 and L. reuteri LA6 isolated from feces of the same human infant were found to produce similar cyclic bacteriocins (named gassericin A and reutericin 6, respectively) that cannot be distinguished by molecular weights or primary amino acid sequences. However, reutericin 6 has a narrower spectrum than gassericin A. In this study, gassericin A inhibited the growth of L. reuteri LA6, but reutericin 6 did not inhibit the growth of L. gasseri LA39. Both bacteriocins caused potassium ion efflux from indicator cells and liposomes, but the amounts of efflux and patterns of action were different. Although circular dichroism spectra of purified bacteriocins revealed that both antibacterial peptides are composed mainly of alpha-helices, the spectra of the bacteriocins did not coincide. The results of D- and L-amino acid composition analysis showed that two residues and one residue of D-Ala were detected among 18 Ala residues of gassericin A and reutericin 6, respectively. These findings suggest that the different D-alanine contents of the bacteriocins may cause the differences in modes of action, amounts of potassium ion efflux, and secondary structures. This is the first report that characteristics of native bacteriocins produced by wild lactobacillus strains having the same structural genes are influenced by a difference in D-amino acid contents in the molecules. PMID- 15128551 TI - Molecular and physiological analysis of the role of osmolyte transporters BetL, Gbu, and OpuC in growth of Listeria monocytogenes at low temperatures. AB - Listeria monocytogenes is a ubiquitous food-borne pathogen found widely distributed in nature as well as an undesirable contaminant in a variety of fresh and processed foods. This ubiquity can be at least partly explained by the ability of the organism to grow at high osmolarity and reduced temperatures, a consequence of its ability to accumulate osmo- and cryoprotective compounds termed osmolytes. Single and multiple deletions of the known osmolyte transporters BetL, Gbu, and OpuC significantly reduce growth at low temperatures. During growth in brain heart infusion broth at 7 degrees C, Gbu and OpuC had a more pronounced role in cryoprotection than did BetL. However, upon the addition of betaine to defined medium, the hierarchy of transporter importance shifted to Gbu > BetL > OpuC. Upon the addition of carnitine, only OpuC appeared to play a role in cryoprotection. Measurements of the accumulated osmolytes showed that betaine is preferred over carnitine, while in the absence of a functional Gbu, carnitine was accumulated to higher levels than betaine was at 7 degrees C. Transcriptional analysis of the genes encoding BetL, Gbu, and OpuC revealed that each transporter is induced to different degrees upon cold shock of L. monocytogenes LO28. Additionally, despite being transcriptionally up-regulated upon cold shock, a putative fourth osmolyte transporter, OpuB (identified by bioinformatic analysis and encoded by lmo1421 and lmo1422), showed no significant contribution to listerial chill tolerance. Growth of the quadruple mutant LO28deltaBCGB (deltabetL deltaopuC deltagbu deltaopuB) was comparable to the that of the triple mutant LO28deltaBCGsoe (deltabetL deltaopuC deltagbu) at low temperatures. Here, we conclude that betaine and carnitine transport upon low temperature exposure is mediated via three osmolyte transporters, BetL, Gbu, and OpuC. PMID- 15128552 TI - Relative neurotoxin gene expression in clostridium botulinum type B, determined using quantitative reverse transcription-PCR. AB - A quantitative reverse transcription-PCR (qRT-PCR) method was developed to monitor the relative expression of the type B botulinum neurotoxin (BoNT/B) gene (cntB) in Clostridium botulinum. The levels of cntB mRNA in five type B strains were accurately monitored by using primers specific for cntB and for the reference gene encoding the 16S rRNA. The patterns and relative expression of cntB were different in the different strains. Except for one of the strains investigated, an increase in cntB expression was observed when the bacteria entered the early stationary growth phase. In the proteolytic strain C. botulinum ATCC 7949, the level of cntB mRNA was four- to fivefold higher than the corresponding levels in the other strains. This was confirmed when we quantified the production of extracellular BoNT/B by an enzyme-linked immunosorbent assay and measured the toxicity of BoNT/B by a mouse bioassay. When the effect of exposure to air on cntB expression was investigated, no decline in the relative expression was observed in spite of an 83% reduction in the viable count based on the initial cell number. Instead, the level of cntB mRNA remained the same. When there was an increase in the sodium nitrite concentration, the bacteria needed a longer adjustment time in the medium before exponential growth occurred. In addition, there was a reduction in the expression of cntB compared to the expression of the 16S rRNA gene at higher sodium nitrite concentrations. This was most obvious in the late exponential growth phase, but at the highest sodium nitrite concentration investigated, 45 ppm, a one- to threefold decline in the cntB mRNA level was observed in all growth phases. PMID- 15128553 TI - Quantitative interaction effects of carbon dioxide, sodium chloride, and sodium nitrite on neurotoxin gene expression in nonproteolytic Clostridium botulinum type B. AB - The effects of carbon dioxide, sodium chloride, and sodium nitrite on type B botulinum neurotoxin (BoNT/B) gene (cntB) expression in nonproteolytic Clostridium botulinum were investigated in a tryptone-peptone-yeast extract (TPY) medium. Various concentrations of these selected food preservatives were studied by using a complete factorial design in order to quantitatively study interaction effects, as well as main effects, on the following responses: lag phase duration (LPD), growth rate, relative cntB expression, and extracellular BoNT/B production. Multiple linear regression was used to set up six statistical models to quantify and predict these responses. All combinations of NaCl and NaNO(2) in the growth medium resulted in a prolonged lag phase duration and in a reduction in the specific growth rate. In contrast, the relative BoNT/B gene expression was unchanged, as determined by the cntB-specific quantitative reverse transcription PCR method. This was confirmed when we measured the extracellular BoNT/B concentration by an enzyme-linked immunosorbent assay. CO(2) was found to have a major effect on gene expression when the cntB mRNA levels were monitored in the mid-exponential, late exponential, and late stationary growth phases. The expression of cntB relative to the expression of the 16S rRNA gene was stimulated by an elevated CO(2) concentration; the cntB mRNA level was fivefold greater in a 70% CO(2) atmosphere than in a 10% CO(2) atmosphere. These findings were also confirmed when we analyzed the extracellular BoNT/B concentration; we found that the concentrations were 27 ng x ml(-1). unit of optical density(-1) in the 10% CO(2) atmosphere and 126 ng x ml(-1). unit of optical density(-1) in the 70% CO(2) atmosphere. PMID- 15128554 TI - Stable isotope fractionation caused by glycyl radical enzymes during bacterial degradation of aromatic compounds. AB - Stable isotope fractionation was studied during the degradation of m-xylene, o xylene, m-cresol, and p-cresol with two pure cultures of sulfate-reducing bacteria. Degradation of all four compounds is initiated by a fumarate addition reaction by a glycyl radical enzyme, analogous to the well-studied benzylsuccinate synthase reaction in toluene degradation. The extent of stable carbon isotope fractionation caused by these radical-type reactions was between enrichment factors (epsilon) of -1.5 and -3.9, which is in the same order of magnitude as data provided before for anaerobic toluene degradation. Based on our results, an analysis of isotope fractionation should be applicable for the evaluation of in situ bioremediation of all contaminants degraded by glycyl radical enzyme mechanisms that are smaller than 14 carbon atoms. In order to compare carbon isotope fractionations upon the degradation of various substrates whose numbers of carbon atoms differ, intrinsic epsilon (epsilon(intrinsic)) were calculated. A comparison of epsilon(intrinsic) at the single carbon atoms of the molecule where the benzylsuccinate synthase reaction took place with compound specific epsilon elucidated that both varied on average to the same extent. Despite variations during the degradation of different substrates, the range of epsilon found for glycyl radical reactions was reasonably narrow to propose that rough estimates of biodegradation in situ might be given by using an average epsilon if no fractionation factor is available for single compounds. PMID- 15128555 TI - Viral activity in two contrasting lake ecosystems. AB - For aquatic systems, especially freshwaters, there is little data on the long term (i.e., >6-month period) and depth-related variability of viruses. In this study, we examined virus-induced mortality of heterotrophic bacteria over a 10 month period and throughout the water column in two lakes of the French Massif Central, the oligomesotrophic Lake Pavin and the eutrophic Lake Aydat. Concurrently, we estimated nonviral mortality through heterotrophic nanoflagellate and ciliate bacterivory. Overall, viral infection parameters were much less variable than bacterial production. We found that the frequency of visibly infected cells (FVIC), estimated using transmission electron microscopy, peaked in both lakes at the end of spring (May to June) and in early autumn (September to October). FVIC values were significantly higher in Lake Pavin (mean [M] = 1.6%) than in Lake Aydat (M = 1.1%), whereas the opposite trend was observed for burst sizes, which averaged 25.7 and 30.2 virus particles bacterium( 1), respectively. We detected no significant depth-related differences in FVIC or burst size. We found that in both lakes the removal of bacterial production by flagellate grazing (M(Pavin) = 37.7%, M(Aydat) = 18.5%) was nearly always more than the production removed by viral lysis (M(Pavin) = 16.2%, M(Aydat) = 19%) or ciliate grazing (M(Pavin) = 2.7%, M(Aydat) = 8.8%). However, at specific times and locations, viral lysis prevailed over protistan grazing, for example, in the anoxic hypolimnion of Lake Aydat. In addition, viral mortality represented a relatively constant mortality source in a bacterial community showing large variations in growth rate and subject to large variations in loss rates from grazers. Finally, although viruses did not represent the main agent of bacterial mortality, our data seem to show that their relative importance was higher in the less productive system. PMID- 15128556 TI - Impact of temperature on the physiological status of a potential bioremediation inoculant, Arthrobacter chlorophenolicus A6. AB - Arthrobacter chlorophenolicus A6 (A6) can degrade large amounts of 4-chlorophenol in soil at 5 and 28 degrees C. In this study, we investigated the effects of temperature on the physiological status of this bacterium in pure culture and in soil. A derivative of A6 tagged with the gfp gene (encoding green fluorescent protein [GFP]) was used to specifically quantify A6 cells in soil. In addition, cyano-ditolyl-tetrazoliumchloride was used to stain GFP-fluorescent cells with an active electron transfer system ("viable cells") whereas propidium iodide (PI) was used to stain cells with damaged membranes ("dead cells"). Another derivative of the strain (tagged with the firefly luciferase gene [luc]) was used to monitor the metabolic activity of the cell population, since the bioluminescence phenotype is dependent on cellular energy reserves. When the cells were incubated in soil at 28 degrees C, the majority were stained with PI, indicating that they had lost their cell integrity. In addition, there was a corresponding decline in metabolic activity and in the ability to be grown in cultures on agar plates after incubation in soil at 28 degrees C, indicating that the cells were dying under those conditions. When the cells were incubated in soil at 5 degrees C, by contrast, the majority of the cells remained intact and a large fraction of the population remained metabolically active. A similar trend towards better cell survival at lower temperatures was found in pure-culture experiments. These results make A. chlorophenolicus A6 a good candidate for the treatment of chlorophenol-contaminated soil in cold climates. PMID- 15128557 TI - Isolation, characterization, and U(VI)-reducing potential of a facultatively anaerobic, acid-resistant Bacterium from Low-pH, nitrate- and U(VI)-contaminated subsurface sediment and description of Salmonella subterranea sp. nov. AB - A facultatively anaerobic, acid-resistant bacterium, designated strain FRCl, was isolated from a low-pH, nitrate- and U(VI)-contaminated subsurface sediment at site FW-024 at the Natural and Accelerated Bioremediation Research Field Research Center in Oak Ridge, Tenn. Strain FRCl was enriched at pH 4.5 in minimal medium with nitrate as the electron acceptor, hydrogen as the electron donor, and acetate as the carbon source. Clones with 16S ribosomal DNA (rDNA) sequences identical to the sequence of strain FRCl were also detected in a U(VI)-reducing enrichment culture derived from the same sediment. Cells of strain FRCl were gram negative motile regular rods 2.0 to 3.4 micro m long and 0.7 to 0.9 microm in diameter. Strain FRCl was positive for indole production, by the methyl red test, and for ornithine decarboxylase; it was negative by the Voges-Proskauer test (for acetylmethylcarbinol production), for urea hydrolysis, for arginine dihydrolase, for lysine decarboxylase, for phenylalanine deaminase, for H(2)S production, and for gelatin hydrolysis. Strain FRCl was capable of using O(2), NO(3)(-), S(2)O(3)(2-), fumarate, and malate as terminal electron acceptors and of reducing U(VI) in the cell suspension. Analysis of the 16S rDNA sequence of the isolate indicated that this strain was 96.4% similar to Salmonella bongori and 96.3% similar to Enterobacter cloacae. Physiological and phylogenetic analyses suggested that strain FRCl belongs to the genus Salmonella and represents a new species, Salmonella subterranea sp. nov. PMID- 15128558 TI - Metal toxicity affects fungal and bacterial activities in soil differently. AB - Although the toxic effect of heavy metals on soil microorganism activity is well known, little is known about the effects on different organism groups. The influence of heavy metal addition on total, bacterial, and fungal activities was therefore studied for up to 60 days in a laboratory experiment using forest soil contaminated with different concentrations of Zn or Cu. The effects of the metals differed between the different activity measurements. During the first week after metal addition, the total activity (respiration rate) decreased by 30% at the highest level of contamination and then remained stable during the 60 days of incubation. The bacterial activity (thymidine incorporation rate) decreased during the first days with the level of metal contamination, resulting in a 90% decrease at the highest level of contamination. Bacterial activity then slowly recovered to values similar to those of the control soil. The recovery was faster when soil pH, which had decreased due to metal addition, was restored to control values by liming. Fungal activity (acetate-in-ergosterol incorporation rate) initially increased with the level of metal contamination, being up to 3 and 7 times higher than that in the control samples during the first week at the highest levels of Zn and Cu addition, respectively. The positive effect of metal addition on fungal activity then decreased, but fungal activity was still higher in contaminated than in control soil after 35 days. This is the first direct evidence that fungal and bacterial activities in soil are differently affected by heavy metals. The different responses of bacteria and fungi to heavy metals were reflected in an increase in the relative fungal/bacterial ratio (estimated using phospholipid fatty acid analysis) with increased metal load. PMID- 15128559 TI - Toxicity caused by hydroxycinnamoyl-coenzyme A thioester accumulation in mutants of Acinetobacter sp. strain ADP1. AB - Hydroxycinnamates, aromatic compounds that play diverse roles in plants, are dissimilated by enzymes encoded by the hca genes in the nutritionally versatile, naturally transformable bacterium Acinetobacter sp. strain ADP1. A key step in the hca-encoded pathway is activation of the natural substrates caffeate, p coumarate, and ferulate by an acyl:coenzyme A (acyl:CoA) ligase encoded by hcaC. As described in this paper, Acinetobacter cells with a knockout of the next enzyme in the pathway, hydroxycinnamoyl-CoA hydratase/lyase (HcaA), are extremely sensitive to the presence of the three natural hydroxycinnamate substrates; Escherichia coli cells carrying a subclone with the hcaC gene are hydroxycinnamate sensitive as well. When the hcaA mutation was combined with a mutation in the repressor HcaR, exposure of the doubly mutated Acinetobacter cells to caffeate, p-coumarate, or ferulate at 10(-6) M totally inhibited the growth of cells. The toxicity of p-coumarate and ferulate to a DeltahcaA strain was found to be a bacteriostatic effect. Although not toxic to wild-type cells initially, the diphenolic caffeate was itself converted to a toxin over time in the absence of cells; the converted toxin was bactericidal. In an Acinetobacter strain blocked in hcaA, a secondary mutation in the ligase (HcaC) suppresses the toxic effect. Analysis of suppression due to the mutation of hcaC led to the development of a positive-selection strategy that targets mutations blocking HcaC. An hcaC mutation from one isolate was characterized and was found to result in the substitution of an amino acid that is conserved in a functionally characterized homolog of HcaC. PMID- 15128560 TI - Novel polyketide synthase from Nectria haematococca. AB - We identified a polyketide synthase (PKS) gene, pksN, from a strain of Nectria haematococca by complementing a mutant unable to synthesize a red perithecial pigment. pksN encodes a 2,106-amino-acid polypeptide with conserved motifs characteristic of type I PKS enzymatic domains: beta-ketoacyl synthase, acyltransferase, duplicated acyl carrier proteins, and thioesterase. The pksN product groups with the Aspergillus nidulans WA-type PKSs involved in conidial pigmentation and melanin, bikaverin, and aflatoxin biosynthetic pathways. Inactivation of pksN did not cause any visible change in fungal growth, asexual sporulation, or ascospore formation, suggesting that it is involved in a specific developmental function. We propose that pksN encodes a novel PKS required for the perithecial red pigment biosynthesis. PMID- 15128561 TI - Survival and dormancy of Mycobacterium avium subsp. paratuberculosis in the environment. AB - The survival of Mycobacterium avium subsp. paratuberculosis was studied by culture of fecal material sampled at intervals for up to 117 weeks from soil and grass in pasture plots and boxes. Survival for up to 55 weeks was observed in a dry fully shaded environment, with much shorter survival times in unshaded locations. Moisture and application of lime to soil did not affect survival. UV radiation was an unlikely factor, but infrared wavelengths leading to diurnal temperature flux may be the significant detrimental component that is correlated with lack of shade. The organism survived for up to 24 weeks on grass that germinated through infected fecal material applied to the soil surface in completely shaded boxes and for up to 9 weeks on grass in 70% shade. The observed patterns of recovery in three of four experiments and changes in viable counts were indicative of dormancy, a hitherto unreported property of this taxon. A dps like genetic element and relA, which are involved in dormancy responses in other mycobacteria, are present in the M. avium subsp. paratuberculosis genome sequence, providing indirect evidence for the existence of physiological mechanisms enabling dormancy. However, survival of M. avium subsp. paratuberculosis in the environment is finite, consistent with its taxonomic description as an obligate parasite of animals. PMID- 15128562 TI - Studies of single-chain antibody expression in quiescent Escherichia coli. AB - Quiescent Escherichia coli cells are generated by overexpressing the Rcd transcript in an hns-205 mutant host. The resulting nongrowing, metabolically active cells were used here to express a single-chain antibody fragment (scFv) in shake flask and fermentor cultures. The expression system is based on two plasmids; one carries the product gene expressed from lambdaP(L) under the control of the cI857 temperature-sensitive repressor, while the second expresses Rcd from lambdaP(R). Shifting the culture from 30 to 42 degrees C induces Rcd expression and product expression simultaneously. Our scFv carried a PelB leader, and 90% of the protein was secreted into the culture supernatant. In a batch culture, the supernatant concentration of scFv in the quiescent-cell culture (optical density at 600 nm [OD(600)] of 3.5) was 37 mg x liter(-1), compared to a maximum of 13 mg x liter(-1) in the control culture (final OD(600) of 20). In a fed-batch fermentor culture, quiescent cells were held at an OD(600) of 20 for 24 h and the extracellular scFv concentration reached a maximum of 150 mg x liter( 1). A control culture with a similar feed reached an OD(600) of 80, but despite the higher density, the extracellular scFv concentration did not exceed 35 mg x liter(-1). Quiescent cells at an OD(600) of 50 exhibited a small decline in the specific product formation rate, but nevertheless, an extracellular scFv concentration of 160 mg x liter(-1) was achieved in 8 h. The rate of extracellular accumulation was 10-fold greater in the quiescent culture than in the control culture. This study demonstrates that it is possible to establish high-density quiescent E. coli cultures that are capable of efficient synthesis, folding, and export of proteins. PMID- 15128563 TI - Genomic diversity of Erwinia carotovora subsp. carotovora and its correlation with virulence. AB - We used genetic and biochemical methods to examine the genomic diversity of the enterobacterial plant pathogen Erwinia carotovora subsp. carotovora. The results obtained with each method showed that E. carotovora subsp. carotovora strains isolated from one ecological niche, potato plants, are surprisingly diverse compared to related pathogens. A comparison of 23 partial mdh sequences revealed a maximum pairwise difference of 10.49% and an average pairwise difference of 2.13%, values which are much greater than the maximum variation (1.81%) and average variation (0.75%) previously reported for Escherichia coli. Pulsed-field gel electrophoresis analysis of I-CeuI-digested genomic DNA revealed seven rrn operons in all E. carotovora subsp. carotovora strains examined except strain WPP17, which had only six copies. We identified 26 I-CeuI restriction fragment length polymorphism patterns and observed significant polymorphism in fragment sizes ranging from 100 to 450 kb for all strains. We detected large plasmids in two strains, including the model strain E. carotovora subsp. carotovora 71. The two least virulent strains had an unusual chromosomal structure, suggesting that a particular pulsotype is correlated with virulence. To compare chromosomal organization of multiple enterobacterial genomes, several genes were mapped onto I-CeuI fragments. We identified portions of the genome that appear to be conserved across enterobacteria and portions that have undergone genome rearrangements. We found that the least virulent strain, WPP17, failed to oxidize cellobiose and was missing several hrp and hrc genes. The unexpected variability among isolates obtained from clonal hosts in one region and in one season suggests that factors other than the host plant, potato, drive the evolution of this common environmental bacterium and key plant pathogen. PMID- 15128564 TI - Particle-based multidimensional multispecies biofilm model. AB - In this paper we describe a spatially multidimensional (two-dimensional [2-D] and three-dimensional [3-D]) particle-based approach for modeling the dynamics of multispecies biofilms growing on multiple substrates. The model is based on diffusion-reaction mass balances for chemical species coupled with microbial growth and spreading of biomass represented by hard spherical particles. Effectively, this is a scaled-up version of a previously proposed individual based biofilm model. Predictions of this new particle-based model were quantitatively compared with those obtained with an established one-dimensional (1-D) multispecies model for equivalent problems. A nitrifying biofilm containing aerobic ammonium and nitrite oxidizers, anaerobic ammonium oxidizers, and inert biomass was chosen as an example. The 2-D and 3-D models generally gave the same results. If only the average flux of nutrients needs to be known, 2-D and 1-D models are very similar. However, the behavior of intermediates, which are produced and consumed in different locations within the biofilm, is better described in 2-D and 3-D models because of the multidirectional concentration gradients. The predictions of 2-D or 3-D models are also different from those of 1-D models for slowly growing or minority species in the biofilm. This aspect is related to the mechanism of biomass spreading or advection implemented in the models and should receive more attention in future experimental studies. PMID- 15128565 TI - Enhancing the thermal tolerance and gastric performance of a microbial phytase for use as a phosphate-mobilizing monogastric-feed supplement. AB - The inclusion of phytase in monogastric animal feed has the benefit of hydrolyzing indigestible plant phytate (myo-inositol 1,2,3,4,5,6-hexakis dihydrogen phosphate) to provide poultry and swine with dietary phosphorus. An ideal phytase supplement should have a high temperature tolerance, allowing it to survive the feed pelleting process, a high specific activity at low pHs, and adequate gastric performance. For this study, the performance of a bacterial phytase was optimized by the use of gene site saturation mutagenesis technology. Beginning with the appA gene from Escherichia coli, a library of clones incorporating all 19 possible amino acid changes and 32 possible codon variations in 431 residues of the sequence was generated and screened for mutants exhibiting improved thermal tolerance. Fourteen single site variants were discovered that retained as much as 10 times the residual activity of the wild-type enzyme after a heated incubation regimen. The addition of eight individual mutations into a single construct (Phy9X) resulted in a protein of maximal fitness, i.e., a highly active phytase with no loss of activity after heating at 62 degrees C for 1 h and 27% of its initial activity after 10 min at 85 degrees C, which was a significant improvement over the appA parental phytase. Phy9X also showed a 3.5-fold enhancement in gastric stability. PMID- 15128566 TI - Nucleic acid amplification strategies for DNA microarray-based pathogen detection. AB - DNA microarray-based screening and diagnostic technologies have long promised comprehensive testing capabilities. However, the potential of these powerful tools has been limited by front-end target-specific nucleic acid amplification. Despite the sensitivity and specificity associated with PCR amplification, the inherent bias and limited throughput of this approach constrain the principal benefits of downstream microarray-based applications, especially for pathogen detection. To begin addressing alternative approaches, we investigated four front end amplification strategies: random primed, isothermal Klenow fragment-based, phi29 DNA polymerase-based, and multiplex PCR. The utility of each amplification strategy was assessed by hybridizing amplicons to microarrays consisting of 70 mer oligonucleotide probes specific for enterohemorrhagic Escherichia coli O157:H7 and by quantitating their sensitivities for the detection of O157:H7 in laboratory and environmental samples. Although nearly identical levels of hybridization specificity were achieved for each method, multiplex PCR was at least 3 orders of magnitude more sensitive than any individual random amplification approach. However, the use of Klenow-plus-Klenow and phi29 polymerase-plus-Klenow tandem random amplification strategies provided better sensitivities than multiplex PCR. In addition, amplification biases among the five genetic loci tested were 2- to 20-fold for the random approaches, in contrast to >4 orders of magnitude for multiplex PCR. The same random amplification strategies were also able to detect all five diagnostic targets in a spiked environmental water sample that contained a 63-fold excess of contaminating DNA. The results presented here underscore the feasibility of using random amplification approaches and begin to systematically address the versatility of these approaches for unbiased pathogen detection from environmental sources. PMID- 15128567 TI - Comparative analysis of the conventional and novel pmo (particulate methane monooxygenase) operons from methylocystis strain SC2. AB - In addition to the conventional pmoA gene (pmoA1) encoding the active site polypeptide of particulate methane monooxygenase, a novel pmoA gene copy (pmoA2) is widely distributed among type II methanotrophs (methane-oxidizing bacteria [MOB]) (M. Tchawa Yimga, P. F. Dunfield, P. Ricke, J. Heyer, and W. Liesack, Appl. Environ. Microbiol. 69:5593-5602, 2003). Here we report that the pmoA1 and pmoA2 gene copies in the type II MOB Methylocystis strain SC2 are each part of a complete pmoCAB gene cluster (pmoCAB1, pmoCAB2). A bacterial artificial chromosome (BAC) library of strain SC2 genomic DNA was constructed, and BAC clones carrying either pmoCAB1 or pmoCAB2 were identified. Comparative sequence analysis showed that these two gene clusters exhibit low levels of identity at both the DNA level (67.4 to 70.9%) and the derived protein level (59.3 to 65.6%). In contrast, the secondary structures predicted for PmoCAB1 and PmoCAB2, as well as the derived transmembrane-spanning regions, are nearly identical. This suggests that PmoCAB2 is, like PmoCAB1, a highly hydrophobic, membrane-associated protein. A total of 190 of the 203 amino acid residues representing a highly conserved consensus sequence of the currently known PmoCAB1 and AmoCAB sequence types could be identified in PmoCAB2. The amoCAB gene cluster encodes ammonia monooxygenase and is evolutionarily related to pmoCAB. Analysis of a set of amino acid residues that allowed differentiation between conventional PmoA and AmoA provided further support for the hypothesis that pmoCAB2 encodes a functional equivalent of PmoCAB1. In experiments in which we used 5' rapid amplification of cDNA ends we identified transcriptional start sites 320 and 177 bp upstream of pmoC1 and pmoC2, respectively. Immediately upstream of the transcriptional start sites of both pmoCAB1 and pmoCAB2, sequence motifs similar to Escherichia coli sigma(70) promoters were identified. PMID- 15128568 TI - Growth of polychlorinated-biphenyl-degrading bacteria in the presence of biphenyl and chlorobiphenyls generates oxidative stress and massive accumulation of inorganic polyphosphate. AB - Inorganic polyphosphate (polyP) plays a significant role in increasing bacterial cell resistance to unfavorable environmental conditions and in regulating different biochemical processes. Using transmission electron microscopy of the polychlorinated biphenyl (PCB)-degrading bacterium Pseudomonas sp. strain B4 grown in defined medium with biphenyl as the sole carbon source, we observed large and abundant electron-dense granules at all stages of growth and following a shift from glucose to biphenyl or chlorobiphenyls. Using energy dispersive X ray analysis and electron energy loss spectroscopy with an integrated energy filtered transmission electron microscope, we demonstrated that these granules were mainly composed of phosphate. Using sensitive enzymatic methods to quantify cellular polyP, we confirmed that this polymer accumulates in PCB-degrading bacteria when they grow in the presence of biphenyl and chlorobiphenyls. Concomitant increases in the levels of the general stress protein GroEl and reactive oxygen species were also observed in chlorobiphenyl-grown cells, indicating that these bacteria adjust their physiology with a stress response when they are confronted with compounds that serve as carbon and energy sources and at the same time are chemical stressors. PMID- 15128569 TI - In vivo study of trichoderma-pathogen-plant interactions, using constitutive and inducible green fluorescent protein reporter systems. AB - Plant tissue colonization by Trichoderma atroviride plays a critical role in the reduction of diseases caused by phytopathogenic fungi, but this process has not been thoroughly studied in situ. We monitored in situ interactions between gfp tagged biocontrol strains of T. atroviride and soilborne plant pathogens that were grown in cocultures and on cucumber seeds by confocal scanning laser microscopy and fluorescence stereomicroscopy. Spores of T. atroviride adhered to Pythium ultimum mycelia in coculture experiments. In mycoparasitic interactions of T. atroviride with P. ultimum or Rhizoctonia solani, the mycoparasitic hyphae grew alongside the pathogen mycelia, and this was followed by coiling and formation of specialized structures similar to hooks, appressoria, and papillae. The morphological changes observed depended on the pathogen tested. Branching of T. atroviride mycelium appeared to be an active response to the presence of the pathogenic host. Mycoparasitism of P. ultimum by T. atroviride occurred on cucumber seed surfaces while the seeds were germinating. The interaction of these fungi on the cucumber seeds was similar to the interaction observed in coculture experiments. Green fluorescent protein expression under the control of host inducible promoters was also studied. The induction of specific Trichoderma genes was monitored visually in cocultures, on plant surfaces, and in soil in the presence of colloidal chitin or Rhizoctonia by confocal microscopy and fluorescence stereomicroscopy. These tools allowed initiation of the mycoparasitic gene expression cascade to be monitored in vivo. PMID- 15128570 TI - Novel forms of structural integration between microbes and a hydrothermal vent gastropod from the Indian Ocean. AB - Here we describe novel forms of structural integration between endo- and episymbiotic microbes and an unusual new species of snail from hydrothermal vents in the Indian Ocean. The snail houses a dense population of gamma-proteobacteria within the cells of its greatly enlarged esophageal gland. This tissue setting differs from that of all other vent mollusks, which harbor sulfur-oxidizing endosymbionts in their gills. The significantly reduced digestive tract, the isotopic signatures of the snail tissues, and the presence of internal bacteria suggest a dependence on chemoautotrophy for nutrition. Most notably, this snail is unique in having a dense coat of mineralized scales covering the sides of its foot, a feature seen in no other living metazoan. The scales are coated with iron sulfides (pyrite and greigite) and heavily colonized by epsilon- and delta proteobacteria, likely participating in mineralization of the sclerites. This novel metazoan-microbial collaboration illustrates the great potential of organismal adaptation in chemically and physically challenging deep-sea environments. PMID- 15128571 TI - Vanadium respiration by Geobacter metallireducens: novel strategy for in situ removal of vanadium from groundwater. AB - Vanadium can be an important contaminant in groundwaters impacted by mining activities. In order to determine if microorganisms of the Geobacteraceae, the predominant dissimilatory metal reducers in many subsurface environments, were capable of reducing vanadium(V), Geobacter metallireducens was inoculated into a medium in which acetate was the electron donor and vanadium(V) was the sole electron acceptor. Reduction of vanadium(V) resulted in the production of vanadium(IV), which subsequently precipitated. Reduction of vanadium(V) was associated with cell growth with a generation time of 15 h. No vanadium(V) was reduced and no precipitate was formed in heat-killed or abiotic controls. Acetate was the most effective of all the electron donors evaluated. When acetate was injected into the subsurface to enhance the growth and activity of Geobacteraceae in an aquifer contaminated with uranium and vanadium, vanadium was removed from the groundwater even more effectively than uranium. These studies demonstrate that G. metallireducens can grow via vanadium(V) respiration and that stimulating the activity of Geobacteraceae, and hence vanadium(V) reduction, can be an effective strategy for in situ immobilization of vanadium in contaminated subsurface environments. PMID- 15128572 TI - Anaerobic nitrogen-fixing consortia consisting of clostridia isolated from gramineous plants. AB - We report here the existence of anaerobic nitrogen-fixing consortia (ANFICOs) consisting of N(2)-fixing clostridia and diverse nondiazotrophic bacteria in nonleguminous plants; we found these ANFICOs while attempting to overcome a problem with culturing nitrogen-fixing microbes from various gramineous plants. A major feature of ANFICOs is that N(2) fixation by the anaerobic clostridia is supported by the elimination of oxygen by the accompanying bacteria in the culture. In a few ANFICOs, nondiazotrophic bacteria specifically induced nitrogen fixation of the clostridia in culture. ANFICOs are widespread in wild rice species and pioneer plants, which are able to grow in unfavorable locations. These results indicate that clostridia are naturally occurring endophytes in gramineous plants and that clostridial N(2) fixation arises in association with nondiazotrophic endophytes. PMID- 15128573 TI - Negative cross-communication among wheat rhizosphere bacteria: effect on antibiotic production by the biological control bacterium Pseudomonas aureofaciens 30-84. AB - Phenazine antibiotic production in the biological control bacterium Pseudomonas aureofaciens 30-84 is regulated in part via the PhzR/PhzI N-acyl homoserine lactone (AHL) system. Previous work showed that a subpopulation of the wheat rhizosphere community positively affected phenazine gene expression in strain 30 84 via AHL signals (E. A. Pierson, D. W. Wood, J. A. Cannon, F. M. Blachere, and L. S. Pierson III, Mol. Plant-Microbe Interact. 11:1078-1084, 1998). In the present work, a second subpopulation, one that negatively affected phenazine gene expression, was identified from this rhizosphere community. Strain 30-84 grown in conditioned medium (CM) from several strains produced lower levels of phenazines (1.5- to 9.3-fold) than control when grown in CM from the strain 30-84I(1)/I(2). Growth of the phzB::lacZ reporter strain 30-84Z in this CM resulted in decreased lacZ expression (4.3- to 9.2-fold) compared to growth of the control strain in CM, indicating that inhibition of phzB occurred at the level of gene expression. Preliminary chemical and biological characterizations suggested that these signals, unlike other identified negative signals, were not extractable in ethyl acetate. Introduction of extra copies of phzR and phzI, but not phzI alone, in trans into strain 30-84Z reduced the negative effect on phzB::lacZ expression. The presence of negative-signal-producing strains in a mixture with strain 30-84 reduced strain 30-84's ability to inhibit the take-all disease pathogen in vitro. Together, the results from the previous work on the positive-signal subpopulation and the present work on the negative-signal subpopulation suggest that cross communication among members of the rhizosphere community and strain 30-84 may control secondary metabolite production and pathogen inhibition. PMID- 15128574 TI - Bifidobacterium lactis DSM 10140: identification of the atp (atpBEFHAGDC) operon and analysis of its genetic structure, characteristics, and phylogeny. AB - The atp operon is highly conserved among eubacteria, and it has been considered a molecular marker as an alternative to the 16S rRNA gene. PCR primers were designed from the consensus sequences of the atpD gene to amplify partial atpD sequences from 12 Bifidobacterium species and nine Lactobacillus species. All PCR products were sequenced and aligned with other atpD sequences retrieved from public databases. Genes encoding the subunits of the F(1)F(0)-ATPase of Bifidobacterium lactis DSM 10140 (atpBEFHAGDC) were cloned and sequenced. The deduced amino acid sequences of these subunits showed significant homology with the sequences of other organisms. We identified specific sequence signatures for the genus Bifidobacterium and for the closely related taxa Bifidobacterium lactis and Bifidobacterium animalis and Lactobacillus gasseri and Lactobacillus johnsonii, which could provide an alternative to current methods for identification of lactic acid bacterial species. Northern blot analysis showed that there was a transcript at approximately 7.3 kb, which corresponded to the size of the atp operon, and a transcript at 4.5 kb, which corresponded to the atpC, atpD, atpG, and atpA genes. The transcription initiation sites of these two mRNAs were mapped by primer extension, and the results revealed no consensus promoter sequences. Phylogenetic analysis of the atpD genes demonstrated that the Lactobacillus atpD gene clustered with the genera Listeria, Lactococcus, Streptococcus, and Enterococcus and that the higher G+C content and highly biased codon usage with respect to the genome average support the hypothesis that there was probably horizontal gene transfer. The acid inducibility of the atp operon of B. lactis DSM 10140 was verified by slot blot hybridization by using RNA isolated from acid-treated cultures of B. lactis DSM 10140. The rapid increase in the level of atp operon transcripts upon exposure to low pH suggested that the ATPase complex of B. lactis DSM 10140 was regulated at the level of transcription and not at the enzyme assembly step. PMID- 15128575 TI - Isolation, characterization, and in situ detection of a novel chemolithoautotrophic sulfur-oxidizing bacterium in wastewater biofilms growing under microaerophilic conditions. AB - We successfully isolated a novel aerobic chemolithotrophic sulfur-oxidizing bacterium, designated strain SO07, from wastewater biofilms growing under microaerophilic conditions. For isolation, the use of elemental sulfur (S(0)), which is the most abundant sulfur pool in the wastewater biofilms, as the electron donor was an effective measure to establish an enrichment culture of strain SO07 and further isolation. 16S rRNA gene sequence analysis revealed that newly isolated strain SO07 was affiliated with members of the genus Halothiobacillus, but it was only distantly related to previously isolated species (89% identity). Strain SO07 oxidized elemental sulfur, thiosulfate, and sulfide to sulfate under oxic conditions. Strain SO07 could not grow on nitrate. Organic carbons, including acetate, propionate, and formate, could not serve as carbon and energy sources. Unlike other aerobic sulfur-oxidizing bacteria, this bacterium was sensitive to NaCl; growth in medium containing more than 150 mM was negligible. In situ hybridization combined with confocal laser scanning microscopy revealed that a number of rod-shaped cells hybridized with a probe specific for strain SO07 were mainly present in the oxic biofilm strata (ca. 0 to 100 micro m) and that they often coexisted with sulfate-reducing bacteria in this zone. These results demonstrated that strain SO07 was one of the important sulfur oxidizing populations involved in the sulfur cycle occurring in the wastewater biofilm and was primarily responsible for the oxidation of H(2)S and S(0) to SO(4)(2-) under oxic conditions. PMID- 15128576 TI - Functional expression of the ectoine hydroxylase gene (thpD) from Streptomyces chrysomallus in Halomonas elongata. AB - The formation of hydroxyectoine in the industrial ectoine producer Halomonas elongata was improved by the heterologous expression of the ectoine hydroxylase gene, thpD, from Streptomyces chrysomallus. The efficient conversion of ectoine to hydroxyectoine was achieved by the concerted regulation of thpD by the H. elongata ectA promoter. PMID- 15128577 TI - Antimicrobial resistance of Enterococcus species isolated from produce. AB - The purpose of this study was to characterize the antibiotic resistance profiles of Enterococcus species isolated from fresh produce harvested in the southwestern United States. Among the 185 Enterococcus isolates obtained, 97 (52%) were Enterococcus faecium, 38 (21%) were Enterococcus faecalis, and 50 (27%) were other Enterococcus species. Of human clinical importance, E. faecium strains had a much higher prevalence of resistance to ciprofloxacin, tetracycline, and nitrofurantoin than E. faecalis. E. faecalis strains had a low prevalence of resistance to antibiotics used to treat E. faecalis infections of both clinical and of agricultural relevance, excluding its intrinsic resistance patterns. Thirty-four percent of the isolates had multiple-drug-resistance patterns, excluding intrinsic resistance. Data on the prevalence and types of antibiotic resistance in Enterococcus species isolated from fresh produce may be used to describe baseline antibiotic susceptibility profiles associated with Enterococcus spp. isolated from the environment. The data collected may also help elucidate the role of foods in the transmission of antibiotic-resistant strains to human populations. PMID- 15128578 TI - pmoA-based analysis of methanotrophs in a littoral lake sediment reveals a diverse and stable community in a dynamic environment. AB - Diversity and community structure of aerobic methane-oxidizing bacteria in the littoral sediment of Lake Constance was investigated by cloning analysis and terminal restriction fragment length polymorphism (T-RFLP) fingerprinting of the pmoA gene. Phylogenetic analysis revealed a high diversity of type I and type II methanotrophs in the oxygenated uppermost centimeter of the sediment. T-RFLP profiles indicated a high similarity between the active methanotrophic community in the oxic layer and the inactive community in an anoxic sediment layer at a 10 cm depth. There were also no major changes in community structure between littoral sediment cores sampled in summer and winter. By contrast, the fingerprint patterns showed substantial differences between the methanotrophic communities of littoral and profundal sediments. PMID- 15128579 TI - Plastic encapsulation of stabilized Escherichia coli and Pseudomonas putida. AB - Escherichia coli and Pseudomonas putida dried in hydroxyectoine or trehalose are shown to be highly resistant to the organic solvents chloroform and acetone, and consequently, they can be encapsulated in a viable form in solid plastic materials. Bacteria are recovered by rehydration after physical disruption of the plastic. P. putida incorporated into a plastic coating of maize seeds was shown to colonize roots efficiently after germination. PMID- 15128580 TI - Transformation of folate-consuming Lactobacillus gasseri into a folate producer. AB - Five genes essential for folate biosynthesis in Lactococcus lactis were cloned on a broad-host-range lactococcal vector and were transferred to the folate auxotroph Lactobacillus gasseri. As a result L. gasseri changed from a folate consumer to a folate producer. This principle can be used to increase folate levels in many fermented food products. PMID- 15128581 TI - Characterization of Nocardia asteroides isolates from different ecological habitats on the basis of repetitive extragenic palindromic-PCR fingerprinting. AB - Thirteen isolates of Nocardia asteroides from both soils and aquatic samples (lake and moat sediments, as well as scum from activated sludge), together with a type strain and two known clinical isolates of this species, were characterized by repetitive extragenic palindromic-PCR fingerprinting with the BOX-A1R primer. The resulting DNA fingerprint patterns proved to be strain specific, and cluster analysis distinguished the soil isolates, the aquatic isolates, and the known strains as being in separate groups. PMID- 15128582 TI - Salinity decreases nitrite reductase gene diversity in denitrifying bacteria of wastewater treatment systems. AB - Investigation of the diversity of nirK and nirS in denitrifying bacteria revealed that salinity decreased the diversity in a nitrate-containing saline wastewater treatment system. The predominant nirS clone was related to nirS derived from marine bacteria, and the predominant nirK clone was related to nirK of the genus ALCALIGENES: PMID- 15128583 TI - Bacteriophage-based genetic system for selection of nonsplicing inteins. AB - A genetic selection system that detects splicing and nonsplicing activities of inteins was developed based on the ability to rescue a T4 phage strain with a conditionally inactive DNA polymerase. This phage defect can be complemented by expression of plasmid-encoded phage RB69 DNA polymerase. Insertion of an intein gene into the active site of the RB69 DNA polymerase gene renders polymerase activity and phage viability dependent on protein splicing. The effectiveness of the system was tested by screening for thermosensitive splicing mutants. Development of genetic systems with the potential of identifying protein splicing inhibitors is a first step towards controlling proliferation of pathogenic microbes harboring inteins in essential proteins. PMID- 15128584 TI - Degradation of polycyclic aromatic hydrocarbons by a newly discovered enteric bacterium, Leclercia adecarboxylata. AB - A bacterial strain, PS4040, capable of degrading polycyclic aromatic hydrocarbons for use as the sole carbon source was isolated from oily-sludge-contaminated soil. The 16S rRNA gene showed 98.8% homology to that of Leclercia adecarboxylata. Comparative molecular typing with the clinical strain of L. adecarboxylata revealed that there were few comigrating and few distinct amplimers among them. PMID- 15128585 TI - Albusin B, a bacteriocin from the ruminal bacterium Ruminococcus albus 7 that inhibits growth of Ruminococcus flavefaciens. AB - An approximately 32-kDa protein (albusin B) that inhibited growth of Ruminococcus flavefaciens FD-1 was isolated from culture supernatants of Ruminococcus albus 7. Traditional cloning and gene-walking PCR techniques revealed an open reading frame (albB) encoding a protein with a predicted molecular mass of 32,168 Da. A BLAST search revealed two homologs of AlbB from the unfinished genome of R. albus 8 and moderate similarity to LlpA, a recently described 30-kDa bacteriocin from Pseudomonas sp. strain BW11M1. PMID- 15128586 TI - Multiplex PCR with 16S rRNA gene-targeted primers of bifidobacterium spp. to identify sources of fecal pollution. AB - Bifidobacteria are one of the most common bacterial types found in the intestines of humans and other animals and may be used as indicators of human fecal pollution. The presence of nine human-related Bifidobacterium species was analyzed in human and animal wastewater samples of different origins by using species-specific primers based on 16S rRNA sequences. Only B. adolescentis and B. dentium were found exclusively in human sewage. A multiplex PCR approach with strain-specific primers was developed. The method showed a sensitivity threshold of 10 cells/ml. This new molecular method could provide useful information for the characterization of fecal pollution sources. PMID- 15128587 TI - Osmotic stress leads to decreased intracellular pH of Listeria monocytogenes as determined by fluorescence ratio-imaging microscopy. AB - Intracellular pH (pH(i)) of Listeria monocytogenes was determined after exposure to NaCl or sorbitol in liquid and solid media (agar). Both compounds decreased pH(i), and recovery on solid medium was impaired compared to that in liquid medium. N,N'-dicyclohexylcarbodiimide abolished pH(i) recovery, and lowering a(w) with glycerol showed no effect on pH(i). PMID- 15128588 TI - Specific PCR assay for a tannin-tolerant selenomonas ruminantium isolate, derived from helicase coding sequences. AB - Sequences from a tannin-tolerant Selenomonas ruminantium isolate (EAT2) that hydrolyzes gallic acid were identified. Two exhibited identity to helicases with a wide phylogenetic distribution. PCR amplification by using primers from one helicase gene detected 2000 to 5000 EAT2 genome equivalents but did not amplify total gastrointestinal microbial DNA of nine other ungulate species. PMID- 15128589 TI - Multiplex PCR assay for detection of bacterial pathogens associated with warm water Streptococcosis in fish. AB - A multiplex PCR-based method was designed for the simultaneous detection of the main pathogens involved in warm-water streptococcosis in fish (Streptococcus iniae, Streptococcus difficilis, Streptococcus parauberis, and Lactococcus garvieae). Each of the four pairs of oligonucleotide primers exclusively amplified the targeted gene of the specific microorganism. The sensitivity of the multiplex PCR using purified DNA was 25 pg for S. iniae, 12.5 pg for S. difficilis, 50 pg for S. parauberis, and 30 pg for L. garvieae. The multiplex PCR assay was useful for the specific detection of the four species of bacteria not only in pure culture but also in inoculated fish tissue homogenates and naturally infected fish. Therefore, this method could be a useful alternative to the culture-based method for the routine diagnosis of warm-water streptococcal infections in fish. PMID- 15128590 TI - Lewis X-containing glycans are specific and potent competitive inhibitors of the binding of ZP3 to complementary sites on capacitated, acrosome-intact mouse sperm. AB - Mammalian fertilization requires a cascade of interactions between sperm and the egg's zona pellucida (ZP). O-linked glycans on mouse glycoprotein ZP3 have been implicated in mediating one step of the fertilization process, the firm adhesion of acrosome-intact sperm to the ZP. Experiments to identify structural requirements of a sperm-binding glycan have demonstrated that a Lewis X (Le(x)) containing glycan (Gal beta 4[Fuc alpha 3]GlcNAc-R) was a potent, competitive inhibitor of in vitro sperm-ZP binding (Johnston et al. J Biol Chem 1998; 273: 1888-1895). However, those experiments did not define the particular step in the fertilization pathway that was blocked. The experiments described herein test the hypothesis that Le(x)-containing glycans are specific, competitive inhibitors of the binding of Alexa Fluor 568 fluorochrome (Alexa(568))-labeled ZP3 to sperm and, thus, bind the same sperm surface sites as ZP3. Dose-response analyses demonstrated that these glycans are potent inhibitors (IC(50) approximately 180 nM), which at saturation, reduced Alexa(568)-ZP3 binding by approximately 70%. A Lewis A (Le(a))-capped glycan (Gal beta 3[Fuc alpha 4]GlcNAc) was also a potent inhibitor (IC(50) approximately 150-200 nM), but at saturation, it reduced Alexa(568)-ZP3 binding by only 30%. In contrast, nonfucosylated glycans with nonreducing GlcNAc beta 4 or Gal beta 4 residues did not compete; neither did sialyl-Le(x) (Neu5Ac alpha 3Gal beta 4[Fuc alpha 3]GlcNAc-Lewis X) nor sulfo Le(x) (3'-O-SO(3)-Lewis X). However, at saturation, Gal alpha 3Gal beta 4GlcNAc beta 3Gal beta 4Glc reduced Alexa(568)-ZP3 binding by approximately 70% but with moderate apparent affinity (IC(50) approximately 3000 nM). Fluorescence microscopy revealed that Alexa(568)-labeled Le(x)-Lac-BSA, Le(a)-Lac-BSA, and ZP3 bound to the same sperm surface domains. However, Le(a)-Lac did not inhibit binding of Alexa(568)-Le(x)-Lac-BSA, and Le(x)-Lac did not inhibit binding of Alexa(568)-Le(a)-Lac-BSA. Finally, Le(x)-Lac and Le(a)-Lac had an additive inhibitory effect on Alexa(568)-ZP3 binding. Thus, Le(x) is a ligand for a major class of ZP3 binding sites on mouse sperm, whereas Le(a) binding defines a different but less-abundant class of sites. PMID- 15128591 TI - Lewis X-containing neoglycoproteins mimic the intrinsic ability of zona pellucida glycoprotein ZP3 to induce the acrosome reaction in capacitated mouse sperm. AB - The binding of zona pellucida (ZP) glycoprotein ZP3 to mouse sperm surface receptors is mediated by protein-carbohydrate interactions. Subsequently, ZP3 induces sperm to undergo the acrosome reaction, an obligatory step in fertilization. We have previously identified Lewis X (Le(x); Gal beta 4[Fuc alpha 3]GlcNAc) as a potent inhibitor of in vitro sperm-ZP binding (Johnston et al. J Biol Chem 1998; 273:1888-1895). This glycan is recognized by approximately 70% of the ZP3 binding sites on capacitated, acrosome-intact mouse sperm, whereas Lewis A (Le(a); Gal beta 3[Fuc alpha 4]GlcNAc) is recognized by most of the remaining sites (Kerr et al. Biol Reprod 2004; 71:770-777). Herein, we test the hypothesis that Le(x)- and Le(a)-containing glycans, when clustered on a neoglycoprotein, bind ZP3 receptors on sperm and induce sperm to undergo the acrosome reaction via the same signaling pathways as ZP3. Results show that a Le(x)-containing neoglycoprotein induced the acrosome reaction in a dose-dependent and capacitation-dependent manner. A Le(a)-containing neoglycoprotein also induced sperm to undergo the acrosome reaction but was less potent than Le(x)-containing neoglycoproteins. In contrast, neoglycoproteins containing beta4-lactosamine (Gal beta 4GlcNAc), Lewis B (Fuc alpha 2Gal beta 3[Fuc alpha 4]GlcNAc), and sialyl Le(x) glycans were inactive, as were four other neoglycoproteins with different nonfucosylated glycans. Consistent with these results, unconjugated Le(x)- and Le(a)-capped glycans were dose-dependent inhibitors, which at saturation, reduced the ZP-induced acrosome reaction by about 60% and 30%, respectively. Experiments utilizing pharmacological inhibitors suggest that induction of the acrosome reaction by solubilized ZP and Le(x)- and Le(a)-containing neoglycoproteins require the same calcium-dependent pathway. However, only the ZP-induced acrosome reaction requires a functional G(i) protein. Thus, Le(x)-containing neoglycoproteins bind to a major class of ZP3 receptors on capacitated sperm. A Le(a)-containing neoglycoprotein binds a second ZP3 receptor but is a less-potent inducer of the acrosome reaction. PMID- 15128592 TI - TRAIL and KILLER are expressed and induce apoptosis in the murine preimplantation embryo. AB - TRAIL (tumor necrosis factor [TNF]-related apoptosis-inducing ligand) and KILLER are a death-inducing ligand and receptor pair that belong to the TNF and TNF receptor superfamilies, respectively. To date, only one apoptosis-inducing TRAIL receptor (murine KILLER [MK]) has been identified in mice, and it is a homologue of human Death Receptor 5. Whereas the expression of other death receptors, such as Fas and TNF receptor 1 have been documented in mammalian preimplantation embryos, no evidence currently demonstrates either the presence or the function of TRAIL and its corresponding death receptor, MK. Using reverse transcription polymerase chain reaction and confocal immunofluorescent microscopy, we found that both TRAIL and MK are expressed from the 1-cell through the blastocyst stage of murine preimplantation embryo development. These proteins are localized mainly at the cell surface from the 1-cell through the morula stage. At the blastocyst stage, both TRAIL and MK exhibit an apical staining pattern in the trophectoderm cells. Finally, using the TUNEL assay, we demonstrated that MK induces apoptosis in blastocysts sensitized to TRAIL via actinomycin D. Taken together, these data are the first to demonstrate the presence and function of TRAIL and MK, a death inducing ligand and its receptor, in mammalian preimplantation embryos. PMID- 15128593 TI - Regulatory roles of leptin at the hypothalamic-hypophyseal axis before and after sexual maturation in cattle. AB - Studies assessed, either directly or indirectly, the role of GnRH in leptin mediated stimulation of LH release in cattle before and after sexual maturation. In experiment 1, the objectives were to determine whether leptin could acutely accelerate the frequency of LH pulses, and putatively GnRH pulses, in prepubertal heifers at different stages of development. In experiment 2, we determined directly whether acute, leptin-mediated increases in LH secretion in the fasted, mature female are accompanied by an increase in GnRH secretion. Ten-month-old prepubertal heifers (experiment 1) fed normal- (n = 5) and restricted-growth (n = 5) diets received three injections of saline or recombinant ovine leptin (oleptin; 0.2 microg/kg body weight, i.v.) at hourly intervals during 5-h experiments conducted every 5 wk until all normal-growth heifers were pubertal. Leptin increased mean concentrations of circulating LH regardless of diet, but pulse characteristics were not altered at any age. In experiment 2, ovariectomized, estradiol-implanted cows (n = 5) were fasted twice for 72 h and treated with either saline or oleptin i.v. (as in experiment 1) on Day 3 of each fast. Leptin increased plasma concentrations of LH and third ventricle cerebrospinal fluid concentrations of GnRH, and increased the amplitude of LH and the size of GnRH pulses, respectively, on Day 3 of fasting compared to saline. Overall, results indicate that leptin is unable to accelerate the pulse generator in heifers at any developmental stage. However, leptin-mediated augmentation of LH concentrations and pulse amplitude in the nutritionally stressed, mature female are associated with modifications in GnRH secretory dynamics. PMID- 15128594 TI - Role of Fas-mediated apoptosis and follicle-stimulating hormone on the developmental capacity of bovine cumulus oocyte complexes in vitro. AB - Follicular atresia is believed to be largely regulated by apoptosis. To further understand how apoptosis can affect cumulus cells and oocytes we have evaluated the incidence and regulation of apoptosis affecting bovine cumulus oocyte complexes in vitro. Expression of components of the Fas signaling pathway was studied in both oocytes and cumulus cells by polymerase chain reaction after reverse transcription, immunoblotting, and indirect immunofluorescence. Furthermore, the Fas signaling pathway was activated in cumulus oocyte complexes with an agonistic anti-Fas antibody during in vitro maturation in the presence or absence of FSH. Viability and incidence of apoptosis in cumulus cells were evaluated by assessing membrane integrity and nuclear morphology. Oocyte nuclear maturation was also analyzed, as well as cleavage rates, blastocyst formation rates, and blastocyst quality, following in vitro fertilization. Fas mRNA and protein were expressed both in oocytes and cumulus cells. FasL protein was found in cumulus cells but could not be detected in oocytes, despite its mRNA expression. Both activation of the Fas pathway and presence of FSH during in vitro maturation increased the incidence of apoptosis in cumulus cells, affecting predominantly the middle and peripheral regions of the cumulus. The observed increase, however, had no effect on the developmental competence of the oocytes. PMID- 15128595 TI - Immunoneutralization of growth differentiation factor 9 reveals it partially accounts for mouse oocyte mitogenic activity. AB - Paracrine factors secreted by oocytes play a pivotal role in promoting early ovarian follicle growth and in defining a morphogenic gradient in antral follicles, yet the exact identities of these oocyte factors remain unknown. This study was conducted to determine the extent to which the mitogenic activity of mouse oocytes can be attributed to growth differentiation factor 9 (GDF9). To do this, specific anti-human GDF9 monoclonal antibodies were generated. Based on epitope mapping and bioassays, a GDF9 neutralizing antibody, mAb-GDF9-53, was characterized with very low cross-reactivity with related transforming growth factor (TGF)beta superfamily members, including BMP15 (also called GDF9B). Pep SPOT epitope mapping showed that mAb-GDF9-53 recognizes a short 4-aa sequence, and three-dimensional peptide modeling suggested that this binding motif lies at the C-terminal fingertip of mGDF9. As predicted by sequence alignments and modeling, the antibody detected recombinant GDF9, but not BMP15 in a Western blot and GDF9 protein in oocyte extract and oocyte-conditioned medium. In a mouse mural granulosa cell (MGC) bioassay, mAb-GDF9-53 completely abolished the mitogenic effects of GDF9, but had no effect on TGFbeta1 or activin A-stimulated MGC proliferation. An unrelated IgG at the same dose had no effect on GDF9 activity. This GDF9 neutralizing antibody was then tested in an established oocyte-secreted mitogen bioassay, where denuded oocytes cocultured with granulosa cells promote cell proliferation in a dose-dependent manner. The mAb-GDF9-53 dose dependently (0-160 microg/ml) decreased the mitogenic activity of oocytes but only by approximately 45% at the maximum dose of mAb. Just 5 microg/ml of mAb GDF9-53 neutralized 90% of recombinant mGDF9 mitogenic activity, but only 15% of oocyte activity. Unlike mAb-GDF9-53, a TGFbeta pan-specific neutralizing antibody did not affect the mitogenic capacity of the oocyte, but completely neutralized TGF beta 1-induced DNA synthesis. This study has characterized a specific GDF9 neutralizing antibody. Our data provide the first direct evidence that the endogenous GDF9 protein is an important oocyte-secreted mitogen, but also show that GDF9 accounts for only part of total oocyte bioactivity. PMID- 15128596 TI - Pre-sertoli specific gene expression profiling reveals differential expression of Ppt1 and Brd3 genes within the mouse genital ridge at the time of sex determination. AB - In mammals, testis determination is initiated when the SRY gene is expressed in pre-Sertoli cells of the undifferentiated genital ridge. SRY directs the differentiation of these cells into Sertoli cells and initiates the testis differentiation pathway via currently ill-defined mechanisms. Because Sertoli cells are the first somatic cells to differentiate within the developing testis, it is likely that the signals for orchestrating testis determination are expressed within pre-Sertoli cells. We have previously generated a transgenic mouse line that expresses green fluorescent protein under the control of the pig SRY promoter, thus marking pre-Sertoli cells via fluorescence. We have now used suppression-subtractive hybridization (SSH) to construct a normalized cDNA library derived from fluorescence-activated cell sorting (FACS) purified pre Sertoli cells taken from 12.0 to 12.5 days postcoitum (dpc) fetal transgenic mouse testes. A total of 35 candidate cDNAs for known genes were identified. Detection of Sf1, a gene known for its role in sex determination as well as Vanin 1, Vcp1, Sparc, and Aldh3a1, four genes previously identified in differential screens as gene overexpressed in developing testis compared with ovary, support the biological validity of our experimental model. Whole-mount in situ hybridization was performed on the 35 candidate genes for qualitative differential expression between male and female genital ridges; six were upregulated in the testis and one was upregulated in the ovary. The expression pattern of two genes, Ppt1 and Brd3, were examined in further detail. We conclude that combining transgenically marked fluorescent cell populations with differential expression screening is useful for cell expression profiling in developmental systems such as sex determination and differentiation. PMID- 15128597 TI - Progress with nonhuman primate embryonic stem cells. AB - Embryonic stem cells hold potential in the fields of regenerative medicine, developmental biology, tissue regeneration, disease pathogenicity, and drug discovery. Embryonic stem (ES) cell lines are now available in primates, including man, rhesus, and cynomologous monkeys. Monkey ES cells serve as invaluable clinically relevant models for studies that can't be conducted in humans because of practical or ethical limitations, or in rodents because of differences in physiology and anatomy. Here, we review the current status of nonhuman primate research with ES cells, beginning with a description of their isolation, characterization, and availability. Substantial limitations still plague the use of primate ES cells, such as their required growth on feeder layers, poor cloning efficiency, and restricted availability. The ability to produce homogenous populations of both undifferentiated as well as differentiated phenotypes is an important challenge, and genetic approaches to achieving these objectives are discussed. Finally, safety, efficiency, and feasibility issues relating to the transplantation of ES-derived cells are considered. PMID- 15128598 TI - Interleukin (IL)-1, IL-6, and IL-8 predict mucosal toxicity of vaginal microbicidal contraceptives. AB - Inflammation of the female reproductive tract increases susceptibility to HIV-1 and other viral infections and, thus, it becomes a serious liability for vaginal products. Excessive release of proinflammatory cytokines may alter the mucosal balance between tissue destruction and repair and be linked to enhanced penetration and replication of viral pathogens upon chemical insult. The present study evaluates four surface-active microbicide candidates, nonoxynol-9 (N-9), benzalkonium chloride (BZK), sodium dodecyl sulfate, and sodium monolaurate for their activity against human sperm and HIV, and their capacity to induce an inflammatory response on human vaginal epithelial cells and by the rabbit vaginal mucosa. Spermicidal and virucidal evaluations ranked N-9 as the most potent compound but were unable to predict the impact of the compounds on vaginal cell viability. Interleukin (IL)-1 release in vitro reflected their cytotoxicity profiles more accurately. Furthermore, IL-1 concentrations in vaginal washings correlated with cumulative mucosal irritation scores after single and multiple applications (P < 0.01), showing BZK as the most damaging agent for the vaginal mucosa. BZK induced rapid cell death, IL-1 release, and IL-6 secretion. The other compounds required either more prolonged or repeated contact with the vaginal epithelium to induce a significant inflammatory reaction. Increased IL-8 levels after multiple applications in vivo identified compounds with the highest cumulative mucosal toxicity (P < 0.01). In conclusion, IL-1, IL-6, and IL-8 in the vaginal secretions are sensitive indicators of compound-induced mucosal toxicity. The described evaluation system is a valuable tool in identifying novel vaginal contraceptive microbicides, selecting out candidates that may enhance, rather than decrease, HIV transmission. PMID- 15128599 TI - Plasminogen activator and serine protease inhibitor-E2 (protease nexin-1) expression by bovine granulosa cells in vitro. AB - Remodeling of the extracellular matrix (ECM) occurs during antral follicle growth, and the plasminogen activators (PA) have been implicated in this process in rodents. In the present study, we measured the expression and secretion of PA and the PA inhibitor protease nexin-1 (SerpinE2) in antral and basal bovine granulosa cells from small (<6 mm), medium (6-8 mm), and large follicles (>8 mm) during 6 days of culture in serum-free medium. Casein zymography revealed that the cells secreted predominantly tissue-type PA (tPA) with urokinase (uPA) being associated mainly with cell lysates, and Western blot demonstrated that the cells secreted SerpinE2. Overall, secreted tPA activity was higher in cultures of cells from small follicles compared with large follicles, and secreted SerpinE2 levels were higher in cultures of cells from large follicles. In cultures of cells from small follicles, secreted tPA levels increased with time of culture for antral but not basal cells, and SerpinE2 levels increased with time for basal but not antral cells. In cultures of granulosa cells from large follicles, tPA activity increased significantly with time of culture, whereas SerpinE2 levels decreased. Cell-associated uPA activity decreased with time in cells from medium and large follicles. Reverse-transcription polymerase chain reaction and Northern blot analysis showed that SerpinE2 secretion was regulated largely at the transcriptional level, whereas tPA secretion was not. The data suggest stage dependent regulation of granulosa cell PA and SerpinE2 production, consistent with a role in ECM remodeling during follicle growth. PMID- 15128601 TI - Control selection strategies in case-control studies of childhood diseases. AB - To address concerns regarding the representativeness of controls in case-control studies, two selection strategies were evaluated in a study of childhood leukemia, which commenced in California in 1995. The authors selected two controls per case: one from among children identified by using computerized birth records and located successfully, the other from a roster of friends; both were matched on demographic factors. Sixty-four birth certificate-friend control pairs were enrolled (n = 128). Additionally, 192 "ideal" controls were selected without tracing from the birth records. Data on parental ages, parental education, mother's reproductive history, and birth weight were obtained from the birth certificates of all 320 subjects. For all variables except birth weight, the differences between birth certificate and ideal controls were smaller than those between friend and ideal controls. None of the differences between birth certificate and ideal controls was significant, whereas two factors were significantly different between friend and ideal controls. These findings suggest that friend controls may be less representative than birth certificate controls. Despite difficulty in tracing and a seemingly low participation rate (49.0% for 560 enrolled birth certificate controls), using birth records to recruit controls appears to provide a representative sample of children and an opportunity to assess the representativeness of controls. PMID- 15128602 TI - Invited commentary: Birth certificates--a best control scenario? PMID- 15128600 TI - Gonadotropin-releasing hormone regulates expression of the DNA damage repair gene, Fanconi anemia A, in pituitary gonadotroph cells. AB - Gonadal function is critically dependant on regulated secretion of the gonadotropin hormones from anterior pituitary gonadotroph cells. Gonadotropin biosynthesis and release is triggered by the binding of hypothalamic GnRH to GnRH receptor expressed on the gonadotroph cell surface. The repertoire of regulatory molecules involved in this process are still being defined. We used the mouse L beta T2 gonadotroph cell line, which expresses both gonadotropin hormones, as a model to investigate GnRH regulation of gene expression and differential display reverse transcription-polymerase chain reaction (RT-PCR) to identify and isolate hormonally induced changes. This approach identified Fanconi anemia a (Fanca), a gene implicated in DNA damage repair, as a differentially expressed transcript. Mutations in Fanca account for the majority of cases of Fanconi anemia (FA), a recessively inherited disease identified by congenital defects, bone marrow failure, infertility, and cancer susceptibility. We confirmed expression and hormonal regulation of Fanca mRNA by quantitative RT-PCR, which showed that GnRH induced a rapid, transient increase in Fanca mRNA. Fanca protein was also acutely upregulated after GnRH treatment of L beta T2 cells. In addition, Fanca gene expression was confined to mature pituitary gonadotrophs and adult mouse pituitary and was not expressed in the immature alpha T3-1 gonadotroph cell line. Thus, this study extends the expression profile of Fanca into a highly specialized endocrine cell and demonstrates hormonal regulation of expression of the Fanca locus. We suggest that this regulatory mechanism may have a crucial role in the GnRH-response mechanism of mature gonadotrophs and perhaps the etiology of FA. PMID- 15128604 TI - Marginal structural models for analyzing causal effects of time-dependent treatments: an application in perinatal epidemiology. AB - Marginal structural models (MSMs) are causal models designed to adjust for time dependent confounding in observational studies of time-varying treatments. MSMs are powerful tools for assessing causality with complicated, longitudinal data sets but have not been widely used by practitioners. The objective of this paper is to illustrate the fitting of an MSM for the causal effect of iron supplement use during pregnancy (time-varying treatment) on odds of anemia at delivery in the presence of time-dependent confounding. Data from pregnant women enrolled in the Iron Supplementation Study (Raleigh, North Carolina, 1997-1999) were used. The authors highlight complexities of MSMs and key issues epidemiologists should recognize before and while undertaking an analysis with these methods and show how such methods can be readily interpreted in existing software packages, including SAS and Stata. The authors emphasize that if a data set with rich information on confounders is available, MSMs can be used straightforwardly to make robust inferences about causal effects of time-dependent treatments/exposures in epidemiologic research. PMID- 15128605 TI - Application of a new statistical method to derive dietary patterns in nutritional epidemiology. AB - Because foods are consumed in combination, it is difficult in observational studies to separate the effects of single foods on the development of diseases. A possible way to examine the combined effect of food intakes is to derive dietary patterns by using appropriate statistical methods. The objective of this study was to apply a new statistical method, reduced rank regression (RRR), that is more flexible and powerful than the classic principal component analysis. RRR can be used efficiently in nutritional epidemiology by choosing disease-specific response variables and determining combinations of food intake that explain as much response variation as possible. The authors applied RRR to extract dietary patterns from 49 food groups, specifying four diabetes-related nutrients and nutrient ratios as responses. Data were derived from a nested German case-control study within the European Prospective Investigation into Cancer and Nutrition Potsdam study consisting of 193 cases with incident type 2 diabetes identified until 2001 and 385 controls. The four factors extracted by RRR explained 93.1% of response variation, whereas the first four factors obtained by principal component analysis accounted for only 41.9%. In contrast to principal component analysis and other methods, the new RRR method extracted a significant risk factor for diabetes. PMID- 15128606 TI - A population-based study of the prevalence of abdominal aortic aneurysms in relation to bone mineral density: the Tromso study. AB - In a population-based study of 2,586 men and 2,806 postmenopausal women aged 55 74 years in Tromso, Norway, in 1994-1995, associations between the prevalence of abdominal aortic aneurysms and bone mineral density were examined. The presence of an abdominal aortic aneurysm was assessed by ultrasonography. The bone mineral density of the forearm was measured by single X-ray absorptiometry. In postmenopausal women aged 55-64 years (nine cases of aneurysm), the adjusted odds ratio for abdominal aortic aneurysm was 0.42 (95% confidence interval: 0.19, 0.95) for each standard-deviation increase in bone mineral density. In other age groups (65-69 years and 70-74 years) including a total of 50 cases, the corresponding odds ratios for abdominal aortic aneurysm were 1.17 and 0.70, respectively. In men aged 55-59 years, based on 45 cases, the odds ratio for abdominal aortic aneurysm was 0.72 (95% confidence interval: 0.50, 1.03). In other age groups (60-64, 65-69, and 70-74 years) including a total of 206 cases, the odds ratios ranged from 1.00 to 1.10. The associations among men (in any age group) and among women older than 64 years were not statistically significant. The authors' main conclusion is that abdominal aortic aneurysms and bone mineral density are not related. However, an association in younger subjects cannot be ruled out. PMID- 15128607 TI - Does job strain increase the risk for coronary heart disease or death in men and women? The Framingham Offspring Study. AB - Conflicting findings in the literature have made the relation between job strain and coronary heart disease (CHD) controversial. The effect of high job strain on the 10-year incidence of CHD and total mortality was examined in men and women participating in the Framingham Offspring Study; 3,039 participants, 1,711 men and 1,328 women, aged 18-77 years, were examined between 1984 and 1987 and followed for 10 years. Measures of job strain, occupational characteristics, and risk factors for CHD were collected at the baseline examination. Before and after controlling for systolic blood pressure, body mass index, cigarette smoking, diabetes, and the total/high density lipoprotein cholesterol ratio in Cox proportional hazards models, the authors found that high job strain was not associated with mortality or incident CHD in either men or women over the follow up period. Contrary to expectation, women with active job strain (high demands high control) had a 2.8-fold increased risk of CHD (95% confidence interval: 1.1, 7.2) compared with women with high job strain (high demands-low control). For men, higher education, personal income, and occupational prestige were related to decreased risk of total mortality and CHD. These findings do not support high job strain as a significant risk factor for CHD or death in men or women. PMID- 15128608 TI - Midlife dietary intake of antioxidants and risk of late-life incident dementia: the Honolulu-Asia Aging Study. AB - Antioxidants have been hypothesized to protect against Alzheimer's disease, but studies conducted in late life have been inconsistent. Risk factors measured in midlife may better predict dementia in late life because they are less affected by the disease process. The authors examined the association of midlife dietary intake of antioxidants to late-life dementia and its subtypes. Data were obtained from the Honolulu-Asia Aging Study, a prospective community-based study of Japanese-American men who were aged 45-68 years in 1965-1968, when a 24-hour dietary recall was administered. The analysis included 2,459 men with complete dietary data who were dementia-free at the first assessment in 1991-1993 and were examined up to two times for dementia between 1991 and 1999. The sample included 235 incident cases of dementia (102 cases of Alzheimer's disease, 38 cases of Alzheimer's disease with contributing cerebrovascular disease, and 44 cases of vascular dementia). Relative risks by quartile of intake were calculated using Cox proportional hazards models with age as the time scale, after adjustment for sociodemographic and lifestyle factors, cardiovascular risk factors, other dietary constituents, and apolipoprotein E e4. Intakes of beta-carotene, flavonoids, and vitamins E and C were not associated with the risk of dementia or its subtypes. This analysis suggests that midlife dietary intake of antioxidants does not modify the risk of late-life dementia or its most prevalent subtypes. PMID- 15128609 TI - A case-control study of serum tocopherol levels and the alpha- to gamma tocopherol ratio in radiographic knee osteoarthritis: the Johnston County Osteoarthritis Project. AB - Tocopherols are lipid-soluble antioxidants that may protect against some conditions of aging. The authors examined associations between radiographic knee osteoarthritis and serum levels of alpha-, delta-, and gamma-tocopherol and the alpha:gamma-tocopherol ratio in African-American and White adults from the Johnston County Osteoarthritis Project (North Carolina, 1991-1997). Two hundred cases with radiographic knee osteoarthritis (Kellgren-Lawrence grades > or = 2) and 200 controls (Kellgren-Lawrence grade 0) were randomly selected and matched by age, ethnicity, and sex. Serum tocopherol levels were measured by high performance liquid chromatography. Conditional logistic regression was used to estimate associations between radiographic knee osteoarthritis and tertiles of each tocopherol measure, independent of confounders. Persons in the highest tertile of the alpha:gamma-tocopherol ratio had half the odds of radiographic knee osteoarthritis as those in the lowest tertile (adjusted odds ratio = 0.5, 95% confidence interval: 0.2, 1.2). This inverse association occurred in all ethnic and sex subgroups, significantly in African Americans and men. Radiographic knee osteoarthritis was inversely associated with serum alpha tocopherol in African Americans and men, positively associated with serum gamma tocopherol in men, and unassociated with serum delta-tocopherol. Associations between radiographic knee osteoarthritis and tocopherol isoforms are complex and may vary by ethnicity and sex. PMID- 15128610 TI - History of diabetes mellitus and risk of prostate cancer in physicians. AB - Some studies have suggested that diabetes mellitus may decrease the risk of prostate cancer because of lower insulin levels. To further investigate the relation between diabetes and prostate cancer, a nested case-control study was conducted within the US Physicians' Health Study. Cases (n = 1,110) had been diagnosed with prostate cancer, confirmed on medical record review, during follow up in 1982-1995. Controls (n = 1,110) were selected randomly from men free of prostate cancer and were matched on age and date of randomization. Information on personal history of diabetes and other diseases, lifestyle habits, and body weight/height was self-reported. Logistic regression analysis showed that the odds ratio for prostate cancer was 0.64 (95% confidence interval (CI): 0.43, 0.95) for men with diabetes, relative to those without the disease, after adjustment for potential confounders. Odds ratio estimates were 0.63 (95% CI: 0.35, 1.14), 0.77 (95% CI: 0.35, 1.72), 0.59 (95% CI: 0.21, 1.66), and 0.59 (95% CI: 0.27, 1.27) for diabetes diagnosed 1-5, 6-10, 11-15, and > or = 16 years prior to prostate cancer diagnosis (p for trend < 0.05). Adjusted odds ratios were 1.44 (95% CI: 0.34, 6.17) for stage A prostate cancer and 0.48 (95% CI: 0.28, 0.83) for stages B-D. Results suggest that history of diabetes may be associated with a decreased risk of prostate cancer, especially late-stage tumors. PMID- 15128611 TI - Physical activity and reduced risk of incident sporadic colorectal adenomas: observational support for mechanisms involving energy balance and inflammation modulation. AB - To investigate the role of physical activity, energy balance, and inflammation on the risk of incident sporadic colorectal adenoma, the authors conducted a community- and colonoscopy-based case-control study (n = 177 cases, n = 228 controls) in Winston-Salem and Charlotte, North Carolina, from 1995 to 1997. Participants reported energy intake by a semiquantitative food frequency questionnaire, daily physical activity levels by a standardized questionnaire, and anthropometrics by self-assessment. The odds ratios for adenomas comparing the highest and lowest quantiles of exposure were 0.63 (95% confidence interval (CI): 0.34, 1.17) for physical activity, 0.80 (95% CI: 0.37, 1.73) for total energy intake, 0.70 (95% CI: 0.37, 1.34) for body mass index, 1.46 (95% CI: 0.73, 2.92) for waist/hip ratio, and 2.40 (95% CI: 1.24, 4.63) for height. For the combined effects of these factors, risk was particularly low for those with higher physical activity and low waist/hip ratio (odds ratio = 0.37, 95% CI: 0.18, 0.75) or shorter stature (odds ratio = 0.32, 95% CI: 0.16, 0.62). The inverse effect of physical activity was apparent only among those not taking nonsteroidal antiinflammatory drugs (odds ratio = 0.49, 95% CI: 0.25, 0.94). These findings add further evidence that physical activity and overall patterns indicating positive energy balance increase the risk of adenoma. Furthermore, the results suggest indirectly that biologic mechanisms related to inflammation may play a role in the beneficial effect of physical activity on the risk of incident adenoma. PMID- 15128612 TI - Paternal smoking and pregnancy loss: a prospective study using a biomarker of pregnancy. AB - Results of studies on paternal smoking and spontaneous abortions have been inconsistent. The authors examined the effect of paternal smoking on the risk of pregnancy loss in a prospective cohort of 526 newly married, nonsmoking, female textile workers in China between 1996 and 1998. Upon stopping contraception, subjects provided daily urine specimens and records of vaginal bleeding for up to 1 year or until clinical pregnancy. Daily urinary human chorionic gonadotropin was assayed to detect conception and early pregnancy losses, and pregnancies were followed to detect clinical spontaneous abortions. Subjects were grouped by the number of cigarettes that husbands reported smoking daily: nonsmokers (group 1, n = 216), fewer than 20 cigarettes (group 2, n = 239), and 20 or more cigarettes (group 3, n = 71). Compared with that for group 1, the adjusted odds ratio of early pregnancy loss of any conception for group 2 was 1.04 (95% confidence interval (CI): 0.67, 1.63) and for group 3 was 1.81 (95% CI: 1.00, 3.29). The adjusted hazard ratio of conception for group 2 was 0.90 (95% CI: 0.70, 1.18) and for group 3 was 0.96 (95% CI: 0.66, 1.39), while the adjusted hazard ratio of clinical pregnancy for group 2 was 0.93 (95% CI: 0.72, 1.20) and for group 3 was 0.78 (95% CI: 0.55, 1.12). The authors conclude that heavy paternal smoking increased the risk of early pregnancy loss through maternal and/or paternal exposure. PMID- 15128613 TI - Substance use and sexual risk: a participant- and episode-level analysis among a cohort of men who have sex with men. AB - Prior reports associating substance use with sexual risk behavior have generally used summary measures and have not adjusted for participants' background levels of substance use. In this 1999-2001 US study (the EXPLORE study), the authors determined whether substance use during sex was independently associated with sexual risk during recent sexual episodes, as reported by 4,295 human immunodeficiency virus-negative men who have sex with men. The main outcome measure was serodiscordant unprotected anal sex (SDUA). The influence of participant-level characteristics was examined by using repeated-measures logistic models. In assessing the influence of episode-level predictors on SDUA, the influence of participant-level characteristics, including 6-month substance use, was removed by using conditional logistic regression, in effect making each participant his own control. The authors also adjusted for partner characteristics. Eleven percent of participants reported heavy alcohol use, 37% used poppers, 19% sniffed cocaine, and 13% used amphetamines. In the participant level analysis, use of poppers, amphetamines, and sniffed cocaine as well as heavy alcohol use in the prior 6 months were independently associated with SDUA. In the conditional analysis, consumption of > or = 6 alcoholic drinks or use of poppers, amphetamines, or sniffed cocaine just before or during sex was independently associated with SDUA. The authors concluded that programs aimed at preventing human immunodeficiency virus transmission should emphasize the influence of substance use during sex on increased risk behavior. PMID- 15128614 TI - The human basis pontis: motor syndromes and topographic organization. AB - Clinical-anatomic correlations were performed in 25 patients with focal infarcts in the basilar pons to determine whether pontine lacunar syndromes conform to discrete clinical entities, and whether there is topographic organization of the motor system within the human basis pontis. Twelve clinical signs were scored on a 6-point scale, neuroimaging lesions were mapped and defined with statistical certainty, and structure-function correlation was performed to develop a topographic map of motor function. Clinical findings ranged from major devastation following extensive lesions (pure motor hemiplegia) to incomplete basilar pontine syndrome and restricted deficits after small focal lesions (ataxic hemiparesis, dysarthria-clumsy hand syndrome, dysarthria-dysmetria and dysarthria-facial paresis). The syndromes are not absolutely discrete, and are distinguished from each other by the relative degree of involvement of each clinical feature. Structure-function correlations indicate that strength is conveyed by the corticofugal fibres destined for the spinal cord, whereas dysmetria results from lesions involving the neurons of the basilar pons that link the ipsilateral cerebral cortex with the contralateral cerebellar hemisphere. Facial movement and articulation are localized to rostral and medial basilar pons; hand coordination is medial and ventral in rostral and mid-pons; and arm function is represented ventral and lateral to the hand. Leg coordination is in the caudal half of the pons, with lateral predominance. Swallowing is dependent upon the integrity of a number of regions in the rostral pons. Gait is in medial and lateral locations throughout the rostral- caudal extent of the pons. Dysmetria ipsilateral to the lesion constitutes a disconnection syndrome, as it occurs when the hemipontine lesion is extensive and interrupts pontocerebellar fibres traversing from the opposite, intact side of the pons. The heterogeneity of manifestations reflects the well-organized topography of motor function in the human basis pontis, in agreement with the anatomic organization of the motor corticopontine projections in the monkey. Higher order impairments including motor neglect, paraphasic errors and pathological laughter result from rostral and medial pontine lesions, and may result from disruption of the pontine component of associative corticopontocerebellar circuits. PMID- 15128615 TI - Elevated white matter myo-inositol in clinically isolated syndromes suggestive of multiple sclerosis. AB - Normal-appearing white matter (NAWM) in established multiple sclerosis has been shown to be abnormal using a variety of magnetic resonance (MR) techniques, including proton MR spectroscopy ((1)H-MRS), although the stage at which these changes first appear is less clear. Using a 1.5 T scanner and single-voxel (1)H MRS [TR 3000 ms, TE 30 ms, point-resolved spectroscopy (PRESS) localization], we determined NAWM metabolite concentrations in 96 patients a mean of 19 weeks (range 12-28 weeks) after onset of a clinically isolated syndrome (CIS) suggestive of multiple sclerosis and in 44 healthy control subjects. Absolute concentrations of N-acetyl-aspartate, total creatine and phosphocreatine (Cr), choline-containing compounds, glutamate plus glutamine, and myo-inositol (Ins) were estimated automatically using the LCModel. Compared with control subjects, the concentration of Ins was elevated in CIS NAWM (mean 3.31 mM, SD 0.86 versus mean 3.82 mM, SD 1.06; P = 0.001). The increase in Ins was also seen in the patient subgroup with abnormal T2-weighted MRI (mean 3.88 mM, SD 1.10; P = 0.001) and in those who satisfied the McDonald criteria for multiple sclerosis (mean 4.04 mM, SD 1.31; P = 0.001). An increase in Cr was also observed in CIS NAWM (P = 0.023), but other metabolites did not significantly differ between the whole CIS group and control subjects. There was no significant correlation between NAWM Ins and T2 lesion load. The early increase in Ins may reflect a process of pathogenic importance in multiple sclerosis NAWM. Follow-up studies will investigate whether the increase in NAWM Ins is of prognostic importance for future relapses and disability. PMID- 15128616 TI - Vestibular-evoked muscle responses in patients with spinal cord injury. AB - The vestibular system was activated by galvanic electrical stimulation in 22 patients with spinal cord injury. Three patients were studied standing and all were studied sitting. Electromyographic responses recorded in soleus (standing patients) and the erectores spinae (all patients) were compared with data from 18 control subjects. The vestibular stimulus polarity and head position were arranged so as to produce excitatory medium latency muscle responses in the controls. Responses in the patient group were present bilaterally, present unilaterally or absent below the level of injury. The amplitude of response recorded in erectores spinae at lumbar levels below the lesion in 21 patients (left and right side responses summed) and five control subjects was positively correlated with American Spinal Injuries Association (ASIA) grade: the smallest amplitudes were found in patients with the most severe impairment (Spearman rank correlation coefficient rs = 0.59; P = 0.002, two-tailed). The latency of response (averaged for both sides) was negatively correlated with ASIA grade in 21 patients: the longest latencies were found in patients with the most severe impairment (rs = -0.57; P < 0.01, two-tailed). Amplitude and latency were negatively correlated (rs = -0.72, P < 0.002, two-tailed). The latencies of responses recorded in the erectores spinae at different vertebral levels were linearly related to the vertical distance from the inion to the recording site in both patient and control groups. The conduction velocities of the spinal pathways activated by vestibular stimulation were 4.6 and 10.4 m/s in patient (recording below lesion) and control groups, respectively. Both clinical status (patients recording below lesion, patients recording above lesion and controls) and distance were significant predictors of latency (general linear model, P < 0.0005). It is concluded that measurement of vestibular-evoked responses could provide information on the level and density of spinal cord lesions. PMID- 15128617 TI - The impact of sex and language dominance on material-specific memory before and after left temporal lobe surgery. AB - Recent findings raised evidence that in early-onset left temporal lobe epilepsy, women show greater functional plasticity for verbal memory than men. In particular, women with lesion- or epilepsy-driven atypical language dominance show an advantage over men. The question asked in this study was whether there is evidence of sex- and language dominance-dependent late, i.e. adult age, plasticity for verbal memory when epilepsy surgery is performed in these patients. Pre- and 1-year postoperative memory performance was evaluated in 169 patients (94 males and 75 females) who underwent left temporal lobe surgery and who had WADA testing of hemispheric language dominance prior to surgery. Verbal memory and figural memory were assessed by list-learning paradigms. According to the Bonn intracarotid amobarbital test (IAT) protocol, patients were categorized into left dominant or atypically dominant (right, incomplete left or right, and bilateral dominant) groups. Results were controlled for the hypothesized sex differences. Thirty-four percent of men and 47% of women displayed patterns of atypical language dominance. Atypical dominance was related to an early onset of epilepsy. Men showed a larger time window for development of atypical dominance but, differently from women, the pattern of atypical dominance was more strictly determined by the age at onset of epilepsy. Atypically dominant women showed better verbal memory than typically dominant women or men. After surgery, right dominant patients had better verbal memory outcome than patients with bilateral or left language dominance who showed significant memory loss. No effect of sex on verbal memory change was found. Figural memory deteriorated in men and improved in women, when they were not left dominant. Seizure outcome had no effect on performance changes. It was concluded that better preserved verbal memory in atypically dominant women before surgery indicates greater benefit from atypical dominance in women than men with regard to the initial damage associated with left hemisphere epilepsy. Later in life, when epilepsy surgery causes additional damage, no such sex difference is observed, indicating that the women's advantage over men is fixed to an early time window in life. Postoperative changes in figural memory suggest dynamics in crowding and suppression patterns. Whether this reflects late plasticity and compensation needs further demonstration. For clinical practice, it is important to note that incomplete right hemisphere and bilateral language dominance do not protect against verbal memory loss after left-sided temporal lobe surgery. PMID- 15128619 TI - Skin denervation in type 2 diabetes: correlations with diabetic duration and functional impairments. AB - Sensory neuropathy is a prominent component of diabetic neuropathy. It is not entirely clear how diabetes influences skin innervation, and whether these changes are correlated with clinical signs and laboratory findings. To investigate these issues, we performed skin biopsies on the distal leg of 38 consecutive type 2 diabetic patients with sensory symptoms in lower limbs (25 males and 13 females, aged 56.2 +/- 9.4 years) and analysed the correlations of intraepidermal nerve fibre (IENF) densities in skin with glycaemic status (duration of diabetes, HbA1C, and fasting and post-prandial glucose levels), and functional parameters of small fibres (warm and cold thresholds) and large fibres (vibratory threshold and parameters of nerve conduction studies). Clinically, 23 patients (60.5%) had signs of small-fibre impairment, and 19 patients (50.0%) had signs of large-fibre impairment. IENF densities were much lower in diabetic patients than in age- and gender-matched controls (1.794 +/- 2.120 versus 9.359 +/- 3.466 fibres/mm, P < 0.0001), and 81.6% (31/38) of diabetic patients had reduced IENF densities. IENF densities were negatively associated with the duration of diabetes (standardized coefficient: -0.422, P = 0.015) by analysis with a multivariate linear regression model. Abnormal results of functional examinations were present in 81.6% (warm threshold), 57.9% (cold threshold), 63.2% (vibratory threshold) and 49% (amplitude of sural sensory action potential) of diabetic patients. Among the three sensory thresholds, the warm threshold temperature had the highest correlation with IENF densities (standardized coefficient: -0.773, P < 0.0001). On nerve conduction studies in lower-limb nerves, there were abnormal responses in 54.1% of sural nerves, and 50.0% of peroneal nerves. Of neurophysiological parameters, the amplitude of the sural sensory action potential had the highest correlation with IENF density (standardized coefficient: 0.739, P < 0.0001). On clinical examination, 15 patients showed no sign of small-fibre impairment, but seven of these patients had reduced IENF densities. In conclusion, small-fibre sensory neuropathy presenting with reduced IENF densities and correlated elevation of warm thresholds is a major manifestation of type 2 diabetes. In addition, the extent of skin denervation increases with diabetic duration. PMID- 15128618 TI - The time course of epidermal nerve fibre regeneration: studies in normal controls and in people with diabetes, with and without neuropathy. AB - We sought to develop and validate a standardized cutaneous nerve regeneration model and to define the rate of epidermal nerve fibre (ENF) regeneration first in healthy control subjects and then in neuropathic and neuropathy-free subjects with diabetes. Next, we assessed the effect of different factors on the rate of nerve fibre regeneration and investigated whether such an approach might offer insight into novel trial designs and outcome measures. All subjects had a standardized topical capsaicin dressing applied to the distal lateral thigh. ENF densities derived from skin biopsies were determined at baseline, after capsaicin treatment and at reinnervation time points. For each subject, the best fit line from post-denervation data was determined and the slope was used as the rate of regeneration. In healthy control subjects, regeneration was correlated with psychophysical sensory testing, electron microscopy studies and immunohistochemistry with alternative axonal membrane markers. Topical capsaicin application produced complete or nearly complete denervation of the epidermis in both control subjects and people with diabetes. The rate of regeneration was associated with the baseline ENF density (P < 0.001), but not age (P = 0.75), gender (P = 0.18), epidermal thickness (P = 0.4) or post-capsaicin treatment density (P = 0.7). ENF regeneration, as determined by recovery of ENF density, occurred at a rate of 0.177 +/- 0.075 fibres/mm/day in healthy control subjects and was significantly reduced in subjects with diabetes (0.074 +/- 0.064, P < 0.001) after adjusting for changes in baseline ENF density. Among subjects with diabetes, the presence of neuropathy was associated with a further reduction in regenerative rate (0.10 +/- 0.07 versus 0.04 +/- 0.03, P = 0.03), though diabetes type (P = 0.7), duration of diabetes (P = 0.3) or baseline glycated haemoglobin (P = 0.6) were not significant. These results have several implications. First, topical capsaicin application can produce a uniform epidermal nerve fibre injury that is safe and well tolerated, and offers an efficient strategy to measure and study nerve regeneration in man. Secondly, using our techniques, reduced rates of nerve regeneration were found in people with diabetes without evidence of neuropathy and indicate that abnormalities in peripheral nerve function are present early in diabetes, before signs or symptoms develop. These results suggest that regenerative neuropathy trials could include non-neuropathic subjects and that trial duration can be dramatically shortened. PMID- 15128620 TI - Right temporal cortex is critical for utilization of melodic contextual cues in a pitch constancy task. AB - Pitch constancy, perceiving the same pitch from tones with differing spectral shapes, requires one to extract the fundamental frequency from two sets of harmonics and compare them. We previously showed this difficult task to be easier when tonal context is present, presumably because the context creates a tonal reference point from which to judge the test tone. The present study assessed the role of the right auditory cortex in using tonal context for pitch judgements. Thirty-six patients with focal brain excisions of the right or left anterior temporal lobe (RT, LT) and 12 matched control participants (NC) made pitch judgements on complex tones that could differ in fundamental frequency and/or spectral shape. This task was performed in isolation and within a melodic context. The RT group showed impairments both on trials in which extraction of pitch from differing spectral shapes was required (different-timbre trials) and when this was not required (same-timbre trials). All groups performed poorly in the isolated condition, but improved with melodic context. Degree of improvement varied in that the LT group performed normally, whereas the RT group was not able to obtain the same amount of facilitation from the melodic context. In particular, melodic context did not facilitate the RT group's performance on different-timbre trials. Excisions within Heschl's gyrus did not affect these results, suggesting that the impairments were due to the removal of the anterior temporal cortex. The results of this study therefore implicate right anterior auditory cortical areas in making pitch judgements relative to tones that were heard previously. We propose that auditory association areas located on the anterior portion of the superior temporal gyrus, an area with connections to frontal regions implicated in working memory, could be involved in holding and integrating tonal information. PMID- 15128621 TI - Electromyographic profiles of gait prior to onset of freezing episodes in patients with Parkinson's disease. AB - Freezing in Parkinson's disease is a severe and disabling problem of unknown aetiology. The aim of this study was to analyse the temporal pattern and the magnitude of the electromyographic activity of the lower limb muscles just before freezing and to compare this with a voluntary stop and ongoing gait. We recruited 11 patients with a mean age of 64.8 years (SD 5.1) and a mean Unified Parkinson Disease Rating Scale (part III--off) score of 29 (SD 7.9). Within a standard 3D gait laboratory setting, surface electromyographic (EMG) data of the tibialis anterior (TA) and gastrocnemius (GS) muscles were collected using a portable EMG module. Patients in the off-phase of the medication cycle performed several trials of normal walking and voluntary stops or were exposed to freezing provoking circumstances. Filtered EMG signals were rectified, smoothed and expressed as a percentage of the gait cycle. EMG onset was determined using a preset threshold, corrected after visual inspection. The magnitude of EMG was calculated by integrating EMG signals (iEMG) over (real) time. To control for the altered timing of activity, iEMG was also normalized for time (iEMGnormt). Analysis of variance of repeated measures analysis showed that significantly abnormal timing occurred in the TA and GS muscles with overall preserved reciprocity. Before freezing, TA swing activity already started prematurely during the pre-swing phase, whereas it was significantly shortened during the actual swing phase. For the GS muscle, a similar pattern of premature activation and termination was found during the stance phase before a freeze. GS activity also showed prolonged bursts of activity during the swing phase, not present during the normal and stop condition. Total iEMG activity of both TA and GS was significantly reduced during the pre-freezing gait cycles. However, when controlling for the altered duration of the bursts, the average iEMGnormt increased, as did the peak EMG in TA. In GS, iEMGnormt was not different in the three conditions. In conclusion, our data show that a consistent pattern of premature timing of TA and GS activity occurred before freezing, which was interpreted as a disturbance of central gait cycle timing. The total amount of EMG activity was reduced in both lower limb muscles due to the shortened time in which the muscles were active. In contrast to GS, activity in TA showed increased amplitudes of the EMG bursts, indicating a compensation strategy of pulling the leg into swing. The observed changes contribute to insufficient forward progression, deceleration and eventually a breakdown of movement. PMID- 15128622 TI - In vivo MRI of brain inflammation in human ischaemic stroke. AB - Inflammation contributes to brain damage caused by ischaemic stroke. Macrophages, as the prevailing inflammatory cell population in stroke lesions, can be visualized using ultrasmall superparamagnetic iron oxide (USPIO) as a cell specific contrast agent for MRI. In this single-centre open-labelled clinical phase II study we tested the potential of USPIO-enhanced MRI for macrophage imaging in human ischaemic stroke lesions. In a series of 10 consecutive patients, USPIO contrast agent was infused at the end of the first week after symptom onset. Two follow-up MRI scans were performed 24-36 h and 48-72 h after infusion. Two distinct components of USPIO-related signal changes were discernible, one associated with blood vessels and one representing parenchymal enhancement. Vessel-associated changes appeared as signal loss on T2/T2*-weighted images and decreased from the first to second scan after USPIO infusion, most likely reflecting a transient blood pool effect of the contrast agent. Conversely, parenchymal enhancement was mainly evident on T1-weighted images, increased over time, and matched with the expected distribution of macrophages. Importantly, USPIO-induced signal alterations throughout differed from signatures of conventional gadolinium-enhanced MRI, thus being independent from breakdown of the blood-brain barrier. We suggest that increasing USPIO-enhancement on T1 weighted images indicates brain infiltration by USPIO-laden macrophages. Thus, USPIO-enhanced MRI may provide an in vivo surrogate marker of cellular inflammation in stroke and other CNS pathologies. PMID- 15128623 TI - Will we ever know if leukoreduction of red blood cells should be performed? PMID- 15128624 TI - BIS use in pediatric anesthesia: where are we? PMID- 15128625 TI - Transfusion of leukoreduced red blood cells may decrease postoperative infections: two meta-analyses of randomized controlled trials. AB - PURPOSE: To evaluate the efficacy and effectiveness of red blood cell leukoreduction in reducing postoperative infection, mortality and cancer recurrence, two meta-analyses of randomized controlled trials (RCTs) were conducted. METHODS: A systematic search of the scientific literature was conducted. The pooled relative risk ratio (RR) of developing an adverse postoperative outcome with either leukoreduced or non-leukoreduced blood was calculated using a random effects model. To better estimate the efficacy of leukoreduction, a second analysis of transfused patients only was conducted. RESULTS: Ten RCTs met inclusion criteria and eight provided separate data for patients randomized and transfused. The mean percentage of patients randomized but not transfused was 34%. For postoperative infection, the overall pooled RR was 0.76 [(95% confidence interval (CI): 0.54-1.08] for the "all patients randomized" analysis. For the "only patients transfused" analysis, the pooled RR became clinically and statistically significant (RR = 0.60 (95% CI: 0.38-0.93). For mortality, the pooled RR for the "all patients randomized" analysis was 0.71 (95% CI: 0.45-1.13) and 0.61 (95% CI: 0.36-1.04) for the "only patients transfused" analysis. When analyzing either all patients randomized or all patients transfused, there was no statistically significant difference in cancer recurrence rates (one study only). CONCLUSION: We demonstrated that patients who were transfused leukoreduced red blood cells might benefit from a decrease in postoperative infections. A decrease in mortality may have been realized if more patients had been enrolled in the various randomized trials. Including all patients randomized, regardless of whether or not they were actually transfused diluted the observed clinical benefit of leukoreduction. PMID- 15128626 TI - Cell salvage does not minimize perioperative allogeneic blood transfusion in abdominal vascular surgery: a systematic review. AB - PURPOSE: To determine whether the use of cell salvage reduces the proportion of patients receiving at least one unit of allogeneic packed red blood cells during the perioperative period of an elective vascular surgery. SOURCE: We identified all relevant articles through the combined use of electronic searches of the MEDLINE and EMBASE databases, the Cochrane library as well as hand searching of all randomized clinical trials and review articles. The electronic search included articles published between 1966 and April 2001. The search included textword searches using "autotransfusion," "cell salvage," "device," or Medical Subject Headings "autologous blood transfusion" or a "randomized controlled trials" filter. PRINCIPAL FINDINGS: Five randomized controlled trials (RCT) were identified involving cell salvage and vascular surgeries. In infra renal abdominal aortic aneurysm surgery the risk ratio (the risk of receiving at least one unit of allogeneic red cells) was 0.37 [95% confidence intervals (CI) of 0.06 to 2.36]. In elective aorto-femoral bypass surgery the risk ratio was 0.97 (95% CI of 0.66 to 1.42). The pooled risk ratio for cell salvage in vascular surgery was 0.67 (95% CI of 0.35 to 1.28). CONCLUSION: Cell salvage, a commonly used technique to recover red cells from the operative field, has been the subject of several studies in vascular surgery. There is insufficient evidence to recommend the routine use of cell salvage in elective abdominal aortic aneurysm and aorto femoral bypass surgeries. A large RCT would elucidate whether cell salvage is effective as a blood conservation technique. PMID- 15128627 TI - Forehead is as sensitive as finger pulse oximetry during general anesthesia. AB - PURPOSE: To compare the performance of a forehead probe to a conventional finger pulse oximetry probe in anesthetized patients. METHODS: Eighteen patients participated in the study. Each probe was connected to a Nellcor N-550 pulse oximeter. Anesthesia was induced and maintained with propofol. After intubation, the patients received air to achieve a steady-state of peripheral arterial oxygen saturation (SpO(2)). Ventilation was interrupted to induce a hypoxic state. As soon as one of the two SpO(2)'s decreased to 90%, the patients' lungs were ventilated with 100% oxygen. To evaluate the performance of the two pulse oximeters, time to the lowest (TL), time of recovery (TR) and lag times to beginning of SpO(2) decrease (Lag) were measured. RESULTS: There were no significant differences in TL and TR between forehead and finger pulse oximetry under normal perfusion conditions during general anesthesia. When the axillary artery was compressed to mimic reduced peripheral perfusion, SpO(2) in the forehead decreased sooner than in the finger during hypoxia. The forehead and finger TLs were similar, however, TR was significantly longer in the finger. CONCLUSION: The forehead SpO(2) sensor can be used as an alternative to the conventional finger sensor during general anesthesia. PMID- 15128628 TI - Ambulatory surgery adult patient selection criteria - a survey of Canadian anesthesiologists. AB - PURPOSE: An increasing number of patients with complex medical problems are now considered suitable for ambulatory surgery. The purpose of this study was to identify the current clinical practice of ambulatory surgical patient selection. METHODS: A standardized questionnaire specifying 30 clinical conditions was sent to all practicing anesthesiologists who are members of the Canadian Anesthesiologists' Society. Recipients were asked to indicate if they would provide ambulatory anesthesia (yes/no answers) for an adult patient with each of those isolated conditions. A 75% agreement was considered a majority opinion. RESULTS: One thousand three hundred thirty-seven questionnaires were sent and 774 replies were received (57.8%). Over 75% of anesthesiologists were willing to include in their selection criteria American Society of Anesthesiologists' (ASA) physical status III, patients with low-grade angina pectoris (AP) and congestive heart failure (CHF), prior myocardial infarction, asymptomatic valvular disease, sleep apnea without use of narcotics, morbid obesity (MO) without co-morbidities, insulin dependent diabetes mellitus and malignant hyperthermia (MH) susceptible patients. Over 75% of responders found ASA IV patients, high grade AP and CHF, sleep apnea with postoperative narcotics, MO with co-morbidities and no patient escort to be unsuitable for ambulatory anesthesia. CONCLUSION: Our survey demonstrated that medical conditions with extreme grades of severity (mild or severe) are associated with majority opinion to proceed or not to proceed with ambulatory surgery. Issues with over 75% agreement reflect the common practice. Similar surveys may form a part of patient selection guidelines development in the future. PMID- 15128629 TI - Continuing medical education should be offered by both e-mail and regular mail: a survey of Ontario anesthesiologists. AB - PURPOSE: In response to the Royal College of Physicians and Surgeons of Canada Maintenance of Certification Program (MainCert), a research project was designed to determine the modality by which practicing anesthesiologists in Ontario would like to undertake their continuing education courses. We wished to explore whether interest in electronic delivery correlates with gender, age, location of practice, appointment to teaching hospitals or number of years in practice. METHODS: Following University of Toronto Ethics approval, a survey questionnaire was sent to 875 anesthesiologists practicing in Ontario. Included with the questionnaire was an offer of a free module to be delivered by e-mail, regular mail or fax that could, upon completion, generate MainCert credits. RESULTS: Of the 875 questionnaires mailed, 413 (47%) were returned. A total of 404 responses, 113 from female (30%) and 291 from male (70%) anesthesiologists, were entered in the database. Three hundred and thirty three respondents requested the module and of these 51% preferred delivery by regular mail, 40% by e-mail and 3% by fax. Chi squared tests showed no significant differences between gender, among age groups, location of practice nor affiliation with university/teaching hospitals. When asked to rate their level of comfort with the Internet on a ten-point scale (1 = low, 10 = high), 59% of respondents indicated a level of 8 or higher. Of those who preferred regular mail, 40% indicated that they were also comfortable with electronic communication. CONCLUSION: It was concluded that both e-mail and regular mail options should be offered to facilitate continuing medical education. PMID- 15128630 TI - In Canada, anesthesiologists are less likely to respond to an electronic, compared to a paper questionnaire. AB - PURPOSE: A randomized unblinded controlled trial was used to assess the utility of electronic questionnaires in a survey of Canadian anesthesiologists. METHODS: Postal or electronic questionnaires were sent between November 2001 and March 2002 to 1,333 anesthesiologists registered with the Canadian Anesthesiologists' Society. The primary outcome measure was the difference in response rates between electronic and postal questionnaires. Secondary outcome measures included a comparison of demographic characteristics, cost, and knowledge and practice regarding prophylactic perioperative beta blockade. RESULTS: The overall response rate was 52%. E-mail participants were half as likely as postal participants to respond to the questionnaire (35% vs 69%, relative risk = 0.51, 95% confidence interval 0.45-0.58). Respondents who provided an e-mail address were younger and more likely to be affiliated with an academic institution. There were no significant differences in responses to knowledge and practice questions. The electronic arm was faster than the postal arm and the cost per reply was one third the cost of the postal arm (2.50 dollars vs 8.02 dollars). CONCLUSIONS: Electronic surveys are a means of acquiring information from a large number of individuals in a rapid, efficient and cost-effective manner. This methodology may be particularly valid and useful in surveys of participants with similar backgrounds and internet access. However the lower response rates achieved as compared with postal surveys indicates a need to use vigilance when generalizing results to a broader population. PMID- 15128631 TI - Full scale computer simulators in anesthesia training and evaluation. AB - PURPOSE: With the advent of competency-based curriculum, technology such as full scale computer simulators have acquired an increasingly important role in anesthesia both in training and evaluation. This article reviews the current role of full scale computer simulators in teaching and evaluation in anesthesia. SOURCE: This review draws from existing anesthesia and medical education literature in order to examine and assess the current role of full scale computer simulators in anesthesia education today. PRINCIPAL FINDINGS: The last decade has witnessed a major increase in the use of full scale computer simulators in anesthesia. Many applications have been found for these simulators including teaching and training, evaluation and research. Despite the increasing use and application of full scale computers in anesthesia in the area of teaching and training, definitive studies evaluating its cost effectiveness, its efficacy compared to traditional training methods or its impact on patient outcome are still pending. Although there is some preliminary evidence of reliability and validity in using the simulator to evaluate clinical competence, development in this area has not progressed enough to justify its use in formal, summative evaluation of competence in anesthesia at this time. CONCLUSIONS: As technology acquires an increasingly important role in medical education, full scale computer simulators represent an exciting potential in anesthesia. However, the full potential and role of simulators in anesthesia is still in development and will require a dovetailing of clinical theory and practice with current research in medical education. PMID- 15128632 TI - Epidural anesthesia may attenuate lipid peroxidation during aorto-femoral surgery. AB - PURPOSE: To determine the effect of epidural anesthesia (EP) on oxygenation of the chronically ischemic limb in patients undergoing aorto-femoral bypass grafting and to assess whether it produces an alteration of lipid peroxidation and antioxidant status following revascularization. METHODS: In this prospective, randomized, single-blinded study 40 ASA II or III patients undergoing elective aorto-femoral bypass grafting were allocated to receive general anesthesia (group GA, n = 20), or epidural + GA (group EP, n = 20) during surgery. Femoral venous blood-gas status, activities of the protecting antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSH-px), glutathione reductase (GSH-rd), glutathione (GSH) and thiobarbituric acid-reactive substances (TBARS) as a marker of lipid peroxidation were determined in blood samples taken from the femoral vein at different intervals before and after revascularization. RESULTS: Before the induction of anesthesia in group EP, femoral venous PO(2) [mean (standard deviation), 95% confidence interval] increased after achieving an adequate level of blockade by EP extending to the dermatomal level of T6-8 [29.32 (4.6), 26.34 32.30 to 36.29 (4.6), 33.37-39.22 mmHg, P < 0.05]. Femoral venous PO(2) was similar in both groups thereafter. In the GA group a significant increase in erythrocyte TBARS was observed immediately after restoration of blood flow when compared with baseline values [221.32 (102), 148.35-294-29 to 337.26 (123) 248.99 425.53 nmol*g(-1) hemoglobin, P < 0.01] but not at any other moment. In the EP group TBARS did not increase throughout the study. Within group comparisons revealed no significant differences in GSH, GSH-px, GSH-rd and SOD. CONCLUSION: In patients with atherosclerotic aorto-iliac occlusive disease EP may possibly attenuate lipid peroxidation following revascularization but has no effect on antioxidant enzyme activities. PMID- 15128633 TI - The bispectral index does not correlate with clinical signs of inhalational anesthesia during sevoflurane induction and arousal in children. AB - PURPOSE: Validation of the bispectral index (BIS) in children requires correlating BIS with several levels of sedation, hypnosis and anesthesia. Our purpose was to compare BIS values with objective assessments of the level of hypnosis in children. We postulated that BIS predicted the level of anesthesia during induction and emergence in children. METHODS: In a prospective observational study, we evaluated the BIS monitor in 87 children (ages: 0.3 to 14 yr) ASA physical status I-II undergoing general surgery under sevoflurane and nitrous oxide. Clinical signs of inhalational anesthesia (CSA), the motor response to surgical incision and signs of arousal were used as indicators of the depth of anesthesia. CSA and BIS measurements were paired every minute during induction and emergence until arousal. RESULTS: CSA scores decreased during induction and increased during emergence (P < 0.001) and correlated with changes in sevoflurane concentrations (r = -0.46; P < 0.001). BIS was associated with changes in CSA scores during induction (r = 0.49; P < 0.01) and emergence (r = 0.62; P < 0.01), but the ranges of individual BIS values overlapped several levels of hypnosis. A BIS value greater than 50 had a positive predictive value of 25% for distinguishing between responders and non-responders to surgical incision. A BIS score equal or greater than 72 had a predictive value of 63% for discriminating between pre-arousal and arousal. CONCLUSIONS: BIS correlates with several levels of hypnosis during inhalational induction and emergence in children, but individual BIS values show large inter-individual variability. The BIS monitor identified the physiological changes associated with arousal and distinguished the effects of preoperative sedation during emergence. The use of movement as an endpoint of hypnosis during surgical stimulation does not correlate with BIS values in children. The large inter-individual variability of BIS at different levels of anesthetic depth may limit the applicability of BIS to pediatric anesthesia. PMID- 15128634 TI - Images in anesthesia: 3D systolic anterior motion of the mitral valve. PMID- 15128635 TI - Prolonged right ventricular failure after relief of cardiac tamponade. AB - PURPOSE: To report a case of severe and fatal cardiac complication following pericardiotomy to relieve a malignant tamponade. Right ventricular (RV) failure was responsible for major hypoxemia and for a persistent shunt through a patent foramen ovale. In the absence of pulmonary embolism and coronary occlusion, possible pathophysiologic mechanisms are discussed. CLINICAL FEATURES: This 53-yr old patient presented with oropharyngeal carcinoma previously treated by chemotherapy. One month later, he showed clinical and echocardiographic signs of cardiac tamponade. He had a circumferential pericardial effusion with complete end-diastolic collapse of the right cavities. After an emergent pericardiotomy, he rapidly presented severe hypoxemia. Transesophageal echocardiography showed an akinetic and dilated right ventricle, paradoxical septal wall motion and a normal left ventricular function. A contrast study revealed a right-to-left shunt. No residual pericardial effusion was detectable. Pulmonary angiography excluded a pulmonary embolism and the coronary angiogram was normal. Troponin Ic was elevated postoperatively and peaked on day two (3.78 micro g x L(-1)). The patient died of refractory shock with persistent intracardiac shunt and RV akinesia on day nine. CONCLUSION: Although pulmonary embolism or thrombus of a coronary vessel are the most common causes of prolonged RV failure after pericardiotomy, other mechanisms may be invoked. The possibility is raised that a rapid increase in RV tension may induce the development of muscular injury and impair coronary blood flow, despite a normal coronary angiogram. These could result in a stunned myocardium and opening of a patent foramen ovale. We hypothesize that gradual decompression of a chronic pericardial effusion might be beneficial in patients at risk. PMID- 15128636 TI - Sevoflurane for interventional neuroradiology procedures is associated with more rapid early recovery than propofol. AB - PURPOSE: Sevoflurane and propofol are both suitable for neuroanesthesia but have not previously been compared as maintenance agents for long duration (one to five hours) procedures. METHODS: Using a multicentre international study protocol, 103 patients were randomized to receive either sevoflurane or propofol for maintenance of anesthesia during interventional neuroradiology procedures. After a standardized induction of anesthesia with propofol, 53 patients received sevoflurane 1 to 3% with 60% nitrous oxide (N(2)O) in oxygen (O(2)), and 50 patients received propofol 4 to 10 mg x kg(-1) x hr(-1) with 60% N(2)O in O(2). Maintenance agents were titrated against systemic arterial blood pressure (baseline mean arterial pressure +/- 20%). Recovery times, changes in sedation, pain, nausea and vomiting and psychomotor function during recovery and use of rescue medication were recorded. RESULTS: The group receiving sevoflurane had a more rapid recovery to spontaneous ventilation, extubation, eye opening and orientation compared to the group receiving propofol (3 vs 4 min, P = 0.01; 5 vs 6 min, P = 0.015; 7 vs 10 min, P < 0.001; 13 vs 17 min, P = 0.028; respectively). Sedation, pain, nausea and vomiting, and psychomotor function scores were similar in the two groups. Use of opioid boluses and vasopressors were similar. CONCLUSION: The use of sevoflurane for maintenance of anesthesia for prolonged neuroradiological procedures is associated with more rapid early recovery than propofol and is associated with similar side effects. Sevoflurane and propofol can both be recommended for these procedures. The clinical benefit of the more rapid recovery with sevoflurane is unknown. PMID- 15128637 TI - Best evidence in critical care medicine: daily interruption of sedative infusions in critically ill patients undergoing mechanical ventilation. PMID- 15128638 TI - Sedation with sufentanil in patients receiving pressure support ventilation has no effects on respiration: a pilot study. AB - PURPOSE: To evaluate the effects of sedation with sufentanil on respiratory drive, respiratory pattern, and gas exchange of critically ill patients during pressure support ventilation. METHODS: In this prospective observational cohort study, we observed 12 adult patients receiving partial ventilatory support for acute respiratory failure. Each subject received a continuous infusion of sufentanil at 0.2 to 0.3 micro g x kg(-1) x hr(-1) to obtain a modified Ramsay sedation score between 2 and 3. In basal conditions and at variable distance from the beginning of the sufentanil infusion (10', 30', 60', 120', 24 hr) we evaluated gas exchange, hemodynamic variables, respiratory rate (RR), tidal volume (TV), respiratory pattern, respiratory drive (P0.1) and inspiratory impedance of the respiratory system [P0.1/TV/inspiratory time (Ti)]. RESULTS: The continuous iv administration of 0.2 to 0.3 micro g x kg(-1) x hr(-1) of sufentanil resulted in the desired level of sedation. No significant heart rate, heart rhythm and blood pressure changes were observed. Sufentanil infusion did not affect TV, minute volume, Ti/inspiratory duty cycle, RR, P0.1, P0.1/TV/Ti and gas exchange did not change significantly over the study period. CONCLUSION: A continuous infusion of sufentanil induces "awake" sedation with no detectable effects on respiratory variables in critically ill patients during partial ventilatory support. PMID- 15128639 TI - Fluid choice for resuscitation of the trauma patient: a review of the physiological, pharmacological, and clinical evidence. AB - PURPOSE: Volume replacement regimens are discussed very emotionally. Interpretation of the literature is difficult due to variations in study design, patient population, target for volume replacement, endpoints, and type of fluids. Meta-analyses may not be very helpful because all kinds of patients and very old studies are included. The principles and options for volume replacement were reviewed exclusively in trauma patients and studies from the literature focusing on this problem were analyzed. SOURCE: Using a MEDLINE search, volume replacement therapy in adult trauma patients published in the English language from 1985 to the end of 2002 were identified and analyzed. Studies on prehospital volume replacement, volume replacement in the emergency area or in the operating room, and volume therapy in trauma intensive care unit patients were included. PRINCIPLE FINDINGS: The age-old crystalloid/colloid controvery has still not been resolved but has been enlarged to a colloid/colloid debate. It is now widely accepted that human albumin could easily be replaced by synthetic colloids for volume replacement in trauma patients. No superiority of a specific volume replacement strategy with regard to outcome was found. However, in several studies outcome was not the major endpoint. Although showing some promising results, the importance of hypertonic solutions for volume replacement in the trauma patient is not yet defined. CONCLUSION: The choice of fluid therapy in trauma patients engenders the most controversy and an examination of the body of literature on this subject results in confusion. It is imperative to continue the search for substances that are effective in avoiding the development of post trauma multi-organ dysfunction syndrome without detrimental side-effects. PMID- 15128640 TI - Estimating the incidence of epidural hematoma - is there enough information? PMID- 15128641 TI - "A taste of honey?" (or not?): international medical graduates in North American medicine. PMID- 15128642 TI - Stellate ganglion block and regional cerebral blood volume and oxygenation. PMID- 15128643 TI - Epidural spread depends on the approach used for posterior lumbar plexus block. PMID- 15128644 TI - Nociception is the root cause of postoperative nausea and vomiting: hypothesis. PMID- 15128646 TI - Possible asymptomatic cerebrospinal fluid leak following successful labour epidural catheter placement. PMID- 15128647 TI - Remifentanil pretreatment for propofol injection pain in children. PMID- 15128648 TI - Maxillary jewelry in a parturient: a new cause for concern. PMID- 15128649 TI - Awake intubation using the GlideScope video laryngoscope: initial experience in four cases. PMID- 15128650 TI - [Accidental puncture of the endotracheal tube during a percutaneous tracheostomy]. PMID- 15128651 TI - An endotracheal tube with a stylet and a styletted endotracheal tube are different. PMID- 15128652 TI - An endotracheal tube with a stylet and a styletted endotracheal tube are different. PMID- 15128654 TI - Nitric oxide is involved in growth regulation and re-orientation of pollen tubes. AB - Nitric oxide (NO) controls diverse functions in many cells and organs of animals. It is also produced in plants and has a variety of effects, but little is known about their underlying mechanisms. In the present study, we have discovered a role for NO in the regulation of pollen tube growth, a fast tip-growing cellular system. Pollen tubes must be precisely oriented inside the anatomically complex female ovary in order to deliver sperm. We hypothesized that NO could play a role in this guidance and tested this hypothesis by challenging the growth of pollen tubes with an external NO point source. When a critical concentration was sensed, the growth rate was reduced and the growth axis underwent a subsequent sharp reorientation, after which normal growth was attained. This response was abrogated in the presence of the NO scavenger CPTIO and affected by drugs interfering in the cGMP signaling pathway. The sensitivity threshold of the response was significantly augmented by sildenafil citrate (SC), an inhibitor of cGMP-specific phosphodiesterases in animals. NO distribution inside pollen tubes was investigated using DAF2-DA and was shown to occur mostly in peroxisomes. Peroxisomes are normally excluded from the tip of pollen tubes and little if any NO is found in the cytosol of that region. Our data indicate that the rate and orientation of pollen tube growth is regulated by NO levels at the pollen tube tip and suggest that this NO function is mediated by cGMP. PMID- 15128655 TI - Increased neuromuscular activity causes axonal defects and muscular degeneration. AB - Before establishing terminal synapses with their final muscle targets, migrating motor axons form en passant synaptic contacts with myotomal muscle. Whereas signaling through terminal synapses has been shown to play important roles in pre and postsynaptic development, little is known about the function of these early en passant synaptic contacts. Here, we show that increased neuromuscular activity through en passant synaptic contacts affects pre- and postsynaptic development. We demonstrate that in zebrafish twister mutants, prolonged neuromuscular transmission causes motor axonal extension and muscular degeneration in a dose dependent manner. Cloning of twister reveals a novel, dominant gain-of-function mutation in the muscle-specific nicotinic acetylcholine receptor alpha-subunit, CHRNA1. Moreover, electrophysiological analysis demonstrates that the mutant subunit increases synaptic decay times, thereby prolonging postsynaptic activity. We show that as the first en passant synaptic contacts form, excessive postsynaptic activity in homozygous embryos severely impedes pre- and postsynaptic development, leading to degenerative defects characteristic of the human slow-channel congenital myasthenic syndrome. By contrast, in heterozygous embryos, transient and mild increase in postsynaptic activity does not overtly affect postsynaptic morphology but causes transient axonal defects, suggesting bi directional communication between motor axons and myotomal muscle. Together, our results provide compelling evidence that during pathfinding, myotomal muscle cells communicate extensively with extending motor axons through en passant synaptic contacts. PMID- 15128656 TI - Opposing inputs by Hedgehog and Brinker define a stripe of hairy expression in the Drosophila leg imaginal disc. AB - The sensory organs of the Drosophila adult leg provide a simple model system with which to investigate pattern-forming mechanisms. In the leg, a group of small mechanosensory bristles is organized into a series of longitudinal rows, a pattern that depends on periodic expression of the hairy gene (h) and the proneural genes achaete (ac) and scute (sc). Expression of ac in longitudinal stripes in prepupal leg discs defines the positions of the mechanosensory bristle rows. The ac/sc expression domains are delimited by the Hairy repressor, which is itself periodically expressed. In order to gain insight into the molecular mechanisms involved in leg sensory organ patterning, we have analyzed a Hedgehog (Hh)- and Decapentaplegic (Dpp)-responsive enhancer of the h gene, which directs expression of h in a narrow stripe in the dorsal leg imaginal disc (the D-h stripe). Our studies suggest that the domain of D-h expression is defined by the overlap of Hh and high-level Dpp signaling. We find that the D-h enhancer consists of a Hh-responsive activation element (HHRE) and a repression element (REPE), which responds to the transcriptional repressor Brinker (Brk). The HHRE directs expression of h in a broad stripe along the anteroposterior (AP) compartment boundary. HHRE-directed expression is refined along the AP and dorsoventral axes by Brk1, acting through the REPE. In D-h-expressing cells, Dpp signaling is required to block Brk-mediated repression. This study elucidates a molecular mechanism for integration of the Hh and Dpp signals, and identifies a novel function for Brk as a repressor of Hh-target genes. PMID- 15128657 TI - Multiple points of interaction between retinoic acid and FGF signaling during embryonic axis formation. AB - Anteroposterior (AP) patterning of the developing CNS is crucial for both regional specification and the timing of neurogenesis. Several important factors are involved in AP patterning, including members of the WNT and FGF growth factor families, retinoic acid receptors, and HOX genes. We have examined the interactions between FGF and retinoic signaling pathways. Blockade of FGF signaling downregulates the expression of members of the RAR signaling pathway, RARalpha, RALDH2 and CYP26. Overexpression of a constitutively active RARalpha2 rescues the effects of FGF blockade on the expression of XCAD3 and HOXB9. This suggests that RARalpha2 is required as a downstream target of FGF signaling for the posterior expression of XCAD3 and HOXB9. Surprisingly, we found that posterior expression of FGFR1 and FGFR4 was dependent on the expression of RARalpha2. Anterior expression was also altered with FGFR1 expression being lost, whereas FGFR4 expression was expanded beyond its normal expression domain. RARalpha2 is required for the expression of XCAD3 and HOXB9, and for the ability of XCAD3 to induce HOXB9 expression. We conclude that RARalpha2 is required at multiple points in the posteriorization pathway, suggesting that correct AP neural patterning depends on a series of mutually interactive feedback loops among FGFs, RARs and HOX genes. PMID- 15128658 TI - Guidance of mesoderm cell migration in the Xenopus gastrula requires PDGF signaling. AB - In vertebrates, PDGFA and its receptor, PDGFRalpha, are expressed in the early embryo. Impairing their function causes an array of developmental defects, but the underlying target processes that are directly controlled by these factors are not well known. We show that in the Xenopus gastrula, PDGFA/PDGFRalpha signaling is required for the directional migration of mesodermal cells on the extracellular matrix of the blastocoel roof. Blocking PDGFRalpha function in the mesoderm does not inhibit migration per se, but results in movement that is randomized and no longer directed towards the animal pole. Likewise, compromising PDGFA function in the blastocoel roof substratum abolishes directionality of movement. Overexpression of wild-type PDGFA, or inhibition of PDGFA both lead to randomized migration, disorientation of polarized mesodermal cells, decreased movement towards the animal pole, and reduced head formation and axis elongation. This is consistent with an instructive role for PDGFA in the guidance of mesoderm migration. PMID- 15128659 TI - Cell-autonomous roles of the ecdysoneless gene in Drosophila development and oogenesis. AB - Steroid signaling underlies developmental processes in animals. Mutations that impair steroidogenesis in the fruit fly Drosophila melanogaster provide tools to dissect steroid hormone action genetically. The widely used temperature-sensitive mutation ecdysoneless(1) (ecd(1)) disrupts production of the steroid hormone ecdysone, and causes developmental and reproductive defects. These defects cannot be satisfactorily interpreted without analysis of the ecd gene. Here, we show that ecd encodes an as yet functionally undescribed protein that is conserved throughout eukaryotes. The ecd(1) conditional allele contains an amino acid substitution, whereas three non-conditional larval lethal mutations result in truncated Ecd proteins. Consistent with its role in steroid synthesis, Ecd is expressed in the ecdysone-producing larval ring gland. However, development of ecd-null early larval lethal mutants cannot be advanced by Ecd expression targeted to the ring gland or by hormone feeding. Cell-autonomous ecd function, suggested by these experiments, is evidenced by the inability of ecd(-) clones to survive within developing imaginal discs. Ecd is also expressed in the ovary, and is required in both the follicle cells and the germline for oocyte development. These defects, induced by the loss of ecd, provide the first direct evidence for a cell-autonomous function of this evolutionarily conserved protein. PMID- 15128660 TI - The RhoGEF Pebble is required for cell shape changes during cell migration triggered by the Drosophila FGF receptor Heartless. AB - The FGF receptor Heartless (HTL) is required for mesodermal cell migration in the Drosophila gastrula. We show that mesoderm cells undergo different phases of specific cell shape changes during mesoderm migration. During the migratory phase, the cells adhere to the basal surface of the ectoderm and exhibit extensive protrusive activity. HTL is required for the protrusive activity of the mesoderm cells. Moreover, the early phenotype of htl mutants suggests that HTL is required for the adhesion of mesoderm cells to the ectoderm. In a genetic screen we identified pebble (pbl) as a novel gene required for mesoderm migration. pbl encodes a guanyl nucleotide exchange factor (GEF) for RHO1 and is known as an essential regulator of cytokinesis. We show that the function of PBL in cell migration is independent of the function of PBL in cytokinesis. Although RHO1 acts as a substrate for PBL in cytokinesis, compromising RHO1 function in the mesoderm does not block cell migration. These data suggest that the function of PBL in cell migration might be mediated through a pathway distinct from RHO1. This idea is supported by allele-specific differences in the expressivity of the cytokinesis and cell migration phenotypes of different pbl mutants. We show that PBL is autonomously required in the mesoderm for cell migration. Like HTL, PBL is required for early cell shape changes during mesoderm migration. Expression of a constitutively active form of HTL is unable to rescue the early cellular defects in pbl mutants, suggesting that PBL is required for the ability of HTL to trigger these cell shape changes. These results provide evidence for a novel function of the Rho-GEF PBL in HTL-dependent mesodermal cell migration. PMID- 15128661 TI - The epithelial-mesenchymal transition of the Drosophila mesoderm requires the Rho GTP exchange factor Pebble. AB - Drosophila pebble (pbl) encodes a Rho-family GTP exchange factor (GEF) required for cytokinesis. The accumulation of high levels of PBL protein during interphase and the developmentally regulated expression of pbl in mesodermal tissues suggested that the primary cytokinetic mutant phenotype might be masking other roles. Using various muscle differentiation markers, we found that Even skipped (EVE) expression in the dorsal mesoderm is greatly reduced in pbl mutant embryos. EVE expression in the dorsalmost mesodermal cells is induced in response to DPP secreted by the dorsal epidermal cells. Further analysis revealed that this phenotype is likely to be a consequence of an earlier defect. pbl mutant mesodermal cells fail to undergo the normal epithelial-mesenchymal transition (EMT) and dorsal migration that follows ventral furrow formation. This phenotype is not a secondary consequence of failed cytokinesis, as it is rescued by a mutant form of pbl that does not rescue the cytokinetic defect. In wild-type embryos, newly invaginated cells at the lateral edges of the mesoderm extend numerous protrusions. In pbl mutant embryos, however, cells appear more tightly adhered to their neighbours and extend very few protrusions. Consistent with the dependence of the mesoderm EMT and cytokinesis on actin organisation, the GTP exchange function of the PBL RhoGEF is required for both processes. By contrast, the N-terminal BRCT domains of PBL are required only for the cytokinetic function of PBL. These studies reveal that a novel PBL-mediated intracellular signalling pathway operates in mesodermal cells during the transition from an epithelial to migratory mesenchymal morphology during gastrulation. PMID- 15128662 TI - Tissue-specific G1-phase cell-cycle arrest prior to terminal differentiation in Dictyostelium. AB - The cell cycle status of developing Dictyostelium cells remains unresolved because previous studies have led to conflicting interpretations. We propose a new model of cell cycle events during development. We observe mitosis of about 50% of the cells between 12 and 18 hours of development. Cellular DNA content profiles obtained by flow cytometry and quantification of extra-chromosomal and chromosomal DNA suggest that the daughter cells have half the chromosomal DNA of vegetative cells. Furthermore, little chromosomal DNA synthesis occurs during development, indicating that no S phase occurs. The DNA content in cells sorted by fluorescent tissue-specific reporters indicates that prespore cells divide before prestalk cells and later encapsulate as G1-arrested spores. Consistent with this, germinating spores have one copy of their chromosomes, as judged by fluorescence in situ hybridization and they replicate their chromosomes before mitosis of the emergent amoebae. The DNA content of mature stalk cells suggests that they also attain a G1 state prior to terminal differentiation. As prestalk cells appear to be in G2 up to 22 hours of development, our data suggest that they divide just prior to stalk formation. Our results suggest tissue-specific regulation of G1 phase cell cycle arrest prior to terminal differentiation in Dictyostelium. PMID- 15128663 TI - The immunoglobulin superfamily in Caenorhabditis elegans and Drosophila melanogaster. PMID- 15128665 TI - Cell cycling through development. AB - Regardless of the species, the development of a multicellular organism requires the precise execution of essential developmental processes including patterning, growth, proliferation and differentiation. The cell cycle, in addition to its role as coordinator of DNA replication and mitosis, is also a coordinator of developmental processes, and is a target of developmental signaling pathways. Perhaps because of its central role during development, the cell cycle mechanism, its regulation and its effects on developing tissues is remarkably complex. It was in this light that the Keystone meeting on the cell cycle and development at Snowbird, Utah in January 2004 was held. PMID- 15128666 TI - dMyc is required for larval growth and endoreplication in Drosophila. AB - Members of the Myc family of proto-oncogenes have long been implicated in regulating proliferation, apoptosis and oncogenesis. Recently, transcriptional and biological studies have suggested a direct role for Myc in regulating growth. We have used dm(4), a new null allele of the Drosophila diminutive (dm) gene, which encodes dMyc on the X chromosome, to investigate a role for dMyc in larval endoreplicating tissues, where cellular growth and DNA replication occur in the absence of cell division. Hemizygous dm(4)/Y mutants arrest as second instar larvae, and fat body nuclei of dm(4)/Y mutants fail to attain normal size and normal levels of DNA, resulting from a reduced frequency of S-phase. Thus, dMyc is required for endoreplication and larval growth. In support of this, dMyc, as well as its antagonist dMnt, are expressed in larval tissues in a pattern consistent with their involvement in regulating endoreplication. Overexpression of dMyc in endoreplicating cells results in dramatic increases in nuclear DNA content and cell and nucleolar size, whereas dMnt overexpression has the opposite effect. BrdU incorporation and Cyclin E protein levels continue to oscillate in dMyc-overexpressing cells, indicating that the normal cell cycle control mechanisms are not disrupted. dMyc driven growth and endoreplication are strongly attenuated when the endocycle is blocked with Cyclin E or the cdk inhibitor p21. By contrast, the ability of dMyc to promote growth and endoreplication is only partly reduced when PI3K activity is blocked, suggesting that they influence distinct growth pathways. Our results indicate that larval growth and endoreplication are coupled processes that, although linked to cell cycle control mechanisms, are regulated by dMyc and dMnt. PMID- 15128667 TI - Patterning the forebrain: FoxA4a/Pintallavis and Xvent2 determine the posterior limit of Xanf1 expression in the neural plate. AB - During early development of the nervous system in vertebrates, expression of the homeobox gene Anf/Hesx1/Rpx is restricted to the anterior neural plate subdomain corresponding to the presumptive forebrain. This expression is essential for normal forebrain development and ectopic expression of Xenopus Anf, Xanf1 (also known as Xanf-1), results in severe forebrain abnormalities. By use of transgenic embryos and a novel bi-colour reporter technique, we have identified a cis regulatory element responsible for transcriptional repression of Xanf1 that defines its posterior expression limit within the neural plate. Using this element as the target in a yeast one-hybrid system, we identified two transcription factors, FoxA4a/Pintallavis and Xvent2 (also known as Xvent-2), which are normally expressed posterior to Xanf1. Overexpression of normal and dominant-negative versions of these factors, as well as inhibition of their mRNA translation by antisense morpholinos, show that they actually function as transcriptional repressors of Xanf1 just behind its posterior expression limit. The extremely high similarity of the identified Anf cis-regulatory sequences in Xenopus, chick and human, indicates that the mechanism restricting posterior expression of Anf in Xenopus is shared among vertebrates. Our findings support Nieuwkoop's activation-transformation model for neural patterning, according to which the entire neurectoderm is initially specified towards an anterior fate, which is later suppressed posteriorly as part of the trunk formation process. PMID- 15128668 TI - Notch signaling patterns Drosophila mesodermal segments by regulating the bHLH transcription factor twist. AB - One of the first steps in embryonic mesodermal differentiation is allocation of cells to particular tissue fates. In Drosophila, this process of mesodermal subdivision requires regulation of the bHLH transcription factor Twist. During subdivision, Twist expression is modulated into stripes of low and high levels within each mesodermal segment. High Twist levels direct cells to the body wall muscle fate, whereas low levels are permissive for gut muscle and fat body fate. We show that Su(H)-mediated Notch signaling represses Twist expression during subdivision and thus plays a critical role in patterning mesodermal segments. Our work demonstrates that Notch acts as a transcriptional switch on mesodermal target genes, and it suggests that Notch/Su(H) directly regulates twist, as well as indirectly regulating twist by activating proteins that repress Twist. We propose that Notch signaling targets two distinct 'Repressors of twist' - the proteins encoded by the Enhancer of split complex [E(spl)C] and the HLH gene extra machrochaetae (emc). Hence, the patterning of Drosophila mesodermal segments relies on Notch signaling changing the activities of a network of bHLH transcriptional regulators, which, in turn, control mesodermal cell fate. Since this same cassette of Notch, Su(H) and bHLH regulators is active during vertebrate mesodermal segmentation and/or subdivision, our work suggests a conserved mechanism for Notch in early mesodermal patterning. PMID- 15128669 TI - A regulatory code for neurogenic gene expression in the Drosophila embryo. AB - Bioinformatics methods have identified enhancers that mediate restricted expression in the Drosophila embryo. However, only a small fraction of the predicted enhancers actually work when tested in vivo. In the present study, co regulated neurogenic enhancers that are activated by intermediate levels of the Dorsal regulatory gradient are shown to contain several shared sequence motifs. These motifs permitted the identification of new neurogenic enhancers with high precision: five out of seven predicted enhancers direct restricted expression within ventral regions of the neurogenic ectoderm. Mutations in some of the shared motifs disrupt enhancer function, and evidence is presented that the Twist and Su(H) regulatory proteins are essential for the specification of the ventral neurogenic ectoderm prior to gastrulation. The regulatory model of neurogenic gene expression defined in this study permitted the identification of a neurogenic enhancer in the distant Anopheles genome. We discuss the prospects for deciphering regulatory codes that link primary DNA sequence information with predicted patterns of gene expression. PMID- 15128670 TI - Wingless eliminates ommatidia from the edge of the developing eye through activation of apoptosis. AB - The Drosophila compound eye is formed by selective recruitment of undifferentiated cells into clusters called ommatidia during late larval and early pupal development. Ommatidia at the edge of the eye, which often lack the full complement of photoreceptors and support cells, undergo apoptosis during mid pupation. We have found that this cell death is triggered by the secreted glycoprotein Wingless, which activates its own expression in peripheral ommatidia via a positive feedback loop. Wingless signaling elevates the expression of the pro-apoptotic factors head involution defective, grim and reaper, which are required for ommatidial elimination. We estimate that approximately 6-8% of the total photoreceptor pool in each eye is removed by this mechanism. In addition, we show that the retinal apoptosis previously reported in apc1 mutants occurs at the same time as the peripheral ommatidial cell death and also depends on head involution defective, grim and reaper. We consider the implications of these findings for eye development and function in Drosophila and other organisms. PMID- 15128671 TI - Quantitative contributions of CtBP-dependent and -independent repression activities of Knirps. AB - The Drosophila Knirps protein is a short-range transcriptional repressor that locally inhibits activators by recruiting the CtBP co-repressor. Knirps also possesses CtBP-independent repression activity. The functional importance of multiple repression activities is not well understood, but the finding that Knirps does not repress some cis-regulatory elements in the absence of CtBP suggested that the co-factor may supply a unique function essential to repress certain types of activators. We assayed CtBP-dependent and -independent repression domains of Knirps in Drosophila embryos, and found that the CtBP independent activity, when provided at higher than normal levels, can repress an eve regulatory element that normally requires CtBP. Dose response analysis revealed that the activity of Knirps containing both CtBP-dependent and independent repression activities is higher than that of the CtBP-independent domain alone. The requirement for CtBP at certain enhancers appears to reflect the need for overall higher levels of repression, rather than a requirement for an activity unique to CtBP. Thus, CtBP contributes quantitatively, rather than qualitatively, to overall repression function. The finding that both repression activities are simultaneously deployed suggests that the multiple repression activities do not function as cryptic 'backup' systems, but that each contributes quantitatively to total repressor output. PMID- 15128672 TI - The role of Mixer in patterning the early Xenopus embryo. AB - The transcription factor VegT, is required in early Xenopus embryos for the formation of both the mesoderm and endoderm germ layers. Inherited as a maternal mRNA localized only in vegetal cells, VegT activates the transcription of a large number of transcription factors, as well as signaling ligands that induce cells in the vegetal mass to form endoderm, and the marginal zone to form mesoderm. It is important now to understand the extent to which transcription factors downstream of VegT play individual, or overlapping, roles in the specification and patterning of the endoderm and mesoderm. In addition, it is important to understand the mechanism that specifies the boundary between endoderm and mesoderm. One of the downstream targets of VegT, the homeodomain protein Mixer, is expressed at high levels at the mesoderm/endoderm boundary at the late blastula stage. We therefore examined its functions by blocking its translation using morpholino oligos. In Mixer-depleted embryos, the expression of many signaling ligands and transcription factors was affected. In particular, we found that the expression of several genes, including several normally expressed in mesoderm, was upregulated. Functional assays of Mixer-depleted vegetal cells showed that they have increased mesoderm-inducing activity. This demonstrates that Mixer plays an essential role in controlling the amount of mesoderm induction by the vegetal cells. PMID- 15128673 TI - moz regulates Hox expression and pharyngeal segmental identity in zebrafish. AB - In vertebrate embryos, streams of cranial neural crest (CNC) cells migrate to form segmental pharyngeal arches and differentiate into segment-specific parts of the facial skeleton. To identify genes involved in specifying segmental identity in the vertebrate head, we screened for mutations affecting cartilage patterning in the zebrafish larval pharynx. We present the positional cloning and initial phenotypic characterization of a homeotic locus discovered in this screen. We show that a zebrafish ortholog of the human oncogenic histone acetyltransferase MOZ (monocytic leukemia zinc finger) is required for specifying segmental identity in the second through fourth pharyngeal arches. In moz mutant zebrafish, the second pharyngeal arch is dramatically transformed into a mirror-image duplicated jaw. This phenotype resembles a similar but stronger transformation than that seen in hox2 morpholino oligo (hox2-MO) injected animals. In addition, mild anterior homeotic transformations are seen in the third and fourth pharyngeal arches of moz mutants. moz is required for maintenance of most hox1-4 expression domains and this requirement probably at least partially accounts for the moz mutant homeotic phenotypes. Homeosis and defective Hox gene expression in moz mutants is rescued by inhibiting histone deacetylase activity with Trichostatin A. Although we find early patterning of the moz mutant hindbrain to be normal, we find a late defect in facial motoneuron migration in moz mutants. Pharyngeal musculature is transformed late, but not early, in moz mutants. We detect relatively minor defects in arch epithelia of moz mutants. Vital labeling of arch development reveals no detectable changes in CNC generation in moz mutants, but later prechondrogenic condensations are mispositioned and misshapen. Mirror-image hox2-dependent gene expression changes in postmigratory CNC prefigure the homeotic phenotype in moz mutants. Early second arch ventral expression of goosecoid (gsc) in moz mutants and in animals injected with hox2 MOs shifts from lateral to medial, mirroring the first arch pattern. bapx1, which is normally expressed in first arch postmigratory CNC prefiguring the jaw joint, is ectopically expressed in second arch CNC of moz mutants and hox2-MO injected animals. Reduction of bapx1 function in wild types causes loss of the jaw joint. Reduction of bapx1 function in moz mutants causes loss of both first and second arch joints, providing functional genetic evidence that bapx1 contributes to the moz-deficient homeotic pattern. Together, our results reveal an essential embryonic role and a crucial histone acetyltransferase activity for Moz in regulating Hox expression and segmental identity, and provide two early targets, bapx1 and gsc, of moz and hox2 signaling in the second pharyngeal arch. PMID- 15128674 TI - Investigating the origins of triploblasty: 'mesodermal' gene expression in a diploblastic animal, the sea anemone Nematostella vectensis (phylum, Cnidaria; class, Anthozoa). AB - Mesoderm played a crucial role in the radiation of the triploblastic Bilateria, permitting the evolution of larger and more complex body plans than in the diploblastic, non-bilaterian animals. The sea anemone Nematostella is a non bilaterian animal, a member of the phylum Cnidaria. The phylum Cnidaria (sea anemones, corals, hydras and jellyfish) is the likely sister group of the triploblastic Bilateria. Cnidarians are generally regarded as diploblastic animals, possessing endoderm and ectoderm, but lacking mesoderm. To investigate the origin of triploblasty, we studied the developmental expression of seven genes from Nematostella whose bilaterian homologs are implicated in mesodermal specification and the differentiation of mesodermal cell types (twist, snailA, snailB, forkhead, mef2, a GATA transcription factor and a LIM transcription factor). Except for mef2, the expression of these genes is largely restricted to the endodermal layer, the gastrodermis. mef2 is restricted to the ectoderm. The temporal and spatial expression of these 'mesoderm' genes suggests that they may play a role in germ layer specification. Furthermore, the predominantly endodermal expression of these genes reinforces the hypothesis that the mesoderm and endoderm of triploblastic animals could be derived from the endoderm of a diploblastic ancestor. Alternatively, we consider the possibility that the diploblastic condition of cnidarians is a secondary simplification, derived from an ancestral condition of triploblasty. PMID- 15128675 TI - The first cell cycle after transfer of somatic cell nuclei in a non-human primate. AB - Production of genetically identical non-human primates through somatic cell nuclear transfer (SCNT) can provide diseased genotypes for research and clarify embryonic stem cell potentials. Understanding the cellular and molecular changes in SCNT is crucial to its success. Thus the changes in the first cell cycle of reconstructed zygotes after nuclear transfer (NT) of somatic cells in the Long tailed Macaque (Macaca fascicularis) were studied. Embryos were reconstructed by injecting cumulus and fibroblasts from M. fascicularis and M. silenus, into enucleated M. fascicularis oocytes. A spindle of unduplicated premature condensed chromosome (PCC spindle) from the donor somatic cell was formed at 2 hours after NT. Following activation, the chromosomes segregated and moved towards the two PCC spindle poles, then formed two nuclei. Twenty-four hours after activation, the first cell division occurred. A schematic of the first cell cycle changes following injection of a somatic cell into an enucleated oocyte is proposed. Ninety-three reconstructed embryos were transferred into 31 recipients, resulting in 7 pregnancies that were confirmed by ultrasound; unfortunately none progressed beyond 60 days. PMID- 15128676 TI - Nitric oxide modulates murine yolk sac vasculogenesis and rescues glucose induced vasculopathy. AB - Nitric oxide (NO) has been demonstrated to mediate events during ovulation, pregnancy, blastocyst invasion and preimplantation embryogenesis. However, less is known about the role of NO during postimplantation development. Therefore, in this study, we explored the effects of NO during vascular development of the murine yolk sac, which begins shortly after implantation. Establishment of the vitelline circulation is crucial for normal embryonic growth and development. Moreover, functional inactivation of the endodermal layer of the yolk sac by environmental insults or genetic manipulations during this period leads to embryonic defects/lethality, as this structure is vital for transport, metabolism and induction of vascular development. In this study, we describe the temporally/spatially regulated distribution of nitric oxide synthase (NOS) isoforms during the three stages of yolk sac vascular development (blood island formation, primary capillary plexus formation and vessel maturation/remodeling) and found NOS expression patterns were diametrically opposed. To pharmacologically manipulate vascular development, an established in vitro system of whole murine embryo culture was employed. During blood island formation, the endoderm produced NO and inhibition of NO (L-NMMA) at this stage resulted in developmental arrest at the primary plexus stage and vasculopathy. Furthermore, administration of a NO donor did not cause abnormal vascular development; however, exogenous NO correlated with increased eNOS and decreased iNOS protein levels. Additionally, a known environmental insult (high glucose) that produces reactive oxygen species (ROS) and induces vasculopathy also altered eNOS/iNOS distribution and induced NO production during yolk sac vascular development. However, administration of a NO donor rescued the high glucose induced vasculopathy, restored the eNOS/iNOS distribution and decreased ROS production. These data suggest that NO acts as an endoderm-derived factor that modulates normal yolk sac vascular development, and decreased NO bioavailability and NO mediated sequela may underlie high glucose induced vasculopathy. PMID- 15128677 TI - Deletion of Vhlh in chondrocytes reduces cell proliferation and increases matrix deposition during growth plate development. AB - The von Hippel Lindau tumor suppressor protein (pVHL) is a component of a ubiquitin ligase that promotes proteolysis of the transcription factor hypoxia inducible-factor 1alpha (HIF1alpha), the key molecule in the hypoxic response. We have used conditional inactivation of murine VHL (Vhlh) in all cartilaginous elements to investigate its role in endochondral bone development. Mice lacking Vhlh in cartilage are viable, but grow slower than control littermates and develop a severe dwarfism. Morphologically, Vhlh null growth plates display a significantly reduced chondrocyte proliferation rate, increased extracellular matrix, and presence of atypical large cells within the resting zone. Furthermore, stabilization of the transcription factor HIF1alpha leads to increased expression levels of HIF1alpha target genes in Vhlh null growth plates. Lastly, newborns lacking both Vhlh and Hif1a genes in growth plate chondrocytes display essentially the same phenotype as Hif1a null single mutant mice suggesting that the Vhlh null phenotype could result, at least in part, from increased activity of accumulated HIF1alpha. This is the first study reporting the novel and intriguing findings that pVHL has a crucial role in endochondral bone development and is necessary for normal chondrocyte proliferation in vivo. PMID- 15128678 TI - Hydrolysis of Man9GlcNAc2 and Man8GlcNAc2 oligosaccharides by a purified alpha mannosidase from Candida albicans. AB - A soluble alpha-mannosidase from Candida albicans was purified to homogeneity by sequential size exclusion, ion exchange, and affinity chromatographies in columns of Sepharose CL6B, DEAE Bio-Gel A, and Concanavalin A Sepharose 4B, respectively. Analytical electrophoresis of the purified preparation in 10% SDS-polyacrylamide gels stained with Coomassie blue revealed a single polypeptide of 43 kDa that was responsible for enzyme activity. The purified enzyme primarily trimmed Man(9)GlcNAc(2) to produce Man(8)GlcNAc(2) isomer B and mannose as a function of time of incubation up to 12 h at 37 degrees C. Prolonged incubation with the enzyme resulted in the accumulation after 24 h of other oligosaccharides corresponding to Man(7)GlcNAc(2) and probably Man(6)GlcNAc(2). These two products were also observed when Man(8)GlcNAc(2) isomer B instead of Man(9)GlcNAc(2) was used as substrate. Other oligosaccharides, such as Man(6)GlcNAc(2)-Asn, Man(5)GlcNAc(2)-Asn, and the alpha1,3- and alpha1,6-linked mannobiosides, were not hydrolyzed at all. These properties are consistent with an alpha1,2 mannosidase that may represent a new member of the glycosylhydrolase family 47. PMID- 15128679 TI - The harmful consequences of elevating the doctor-patient relationship to be a primary goal of the general practice consultation. PMID- 15128680 TI - 'Only connect': the centrality of doctor-patient relationships in primary care. PMID- 15128681 TI - Antibiotics for acute respiratory tract symptoms: patients' expectations, GPs' management and patient satisfaction. AB - BACKGROUND: GPs often assume that prescribing antibiotics increases patient satisfaction. OBJECTIVE: The purpose of this study was to determine the associations between receiving antibiotics and information/reassurance on the one hand and patients' satisfaction on the other in patients with acute respiratory tract symptoms, and to assess whether this relationship is influenced by patients' expectations. METHODS: Questionnaires were distributed among patients presenting with acute respiratory tract symptoms in 51 general practices (122 GPs) in the region of Utrecht, The Netherlands. Outcome measures were patients' expectations, GPs' management regarding antibiotic prescribing and information/reassurance, and patient satisfaction. RESULTS: Information/reassurance was expected by 90% of the respondents (916/1014); 97% of those actually received it (888/916). Fifty percent expected antibiotics (467/926), while 73% of them received antibiotics (343/467). Receiving information/reassurance was more strongly associated with satisfaction than an antibiotic prescription [adjusted odds ratio (OR) 10.6; 95% confidence interval (CI) 5.6-20.1, and adjusted OR 2.2; 95% CI 1.3-3.8, respectively]. Patients' expectations towards antibiotics modified these associations; in patients not expecting antibiotics, receiving information/reassurance was the only independent determinant of satisfaction (adjusted OR 21.6; 95% CI 7.4-62.7) and in patients who expected antibiotics, actually receiving antibiotics and receiving information/reassurance were equally important determinants of satisfaction (adjusted OR 3.8; 95% CI 1.9-7.5, and adjusted OR 4.7; 95% CI 1.9-11.9, respectively). CONCLUSION: GPs should first explore patients' expectations about antibiotic treatment before giving information about the self-limitedness of respiratory tract symptoms and (in)effectiveness of antibiotics in order to enhance shared decision making and rationalize antibiotic prescribing. PMID- 15128682 TI - Empirical treatment followed by a test-and-treat strategy is more cost-effective in comparison with prompt endoscopy or radiography in patients with dyspeptic symptoms: a randomized trial in a primary care setting. AB - OBJECTIVE: Management of patients with dyspepsia remains controversial. No consensus has yet been reached concerning diagnostic and medical strategies. We conducted a randomized trial to assess the effectiveness of three management strategies for patients with uninvestigated persistent dyspeptic symptoms. METHODS: A total of 199 patients presenting in primary care with dyspeptic symptoms (age 18-65 years, no alarming symptoms) were randomized to either empirical treatment with omeprazole and, in the case of symptomatic relapse, serological Helicobacter pylori infection testing plus eradication therapy (treat and-test group), prompt upper gastrointestinal endoscopy (endoscopy group) or prompt upper gastrointestinal radiography (radiography group) followed by directed medical treatment. Symptoms, patients' satisfaction and use of resources were recorded during 6 months of follow-up. RESULTS: Sixty-nine patients were assigned to the treat-and-test group, 64 to the radiography group and 66 to the endoscopy group. The median age was 44 years; 104 patients were male and 37% were H.pylori infected. A total of 170 patients (85%) returned the 6 months questionnaire. The numbers of patients with complete symptom relief in the treat and-test group, endoscopy group and radiography group were 21, 16 and 15, respectively, at 3 months (P = 0.59), and 23, 13 and 12, respectively, at 6 months (P = 0.05). Twenty-two patients in the treat-and-test group underwent endoscopy or radiography. Two patients in the endoscopy group and four patients in the radiography group underwent more than one diagnostic test. The average medical cost per patient for the treat-and-test group was euro 276, for the endoscopy group euro 426 and for the radiography group euro 321, respectively. CONCLUSION: Empirical treatment followed by a test-and-eradicate strategy resulted in fewer diagnostic tests, more symptom relief and lower medical costs compared with prompt upper gastrointestinal radiography or endoscopy in the management of uninvestigated patients with persistent dyspeptic symptoms. PMID- 15128683 TI - Prescribing of potentially inappropriate medications to elderly people. AB - OBJECTIVE: To estimate the prevalence and predictors of medications deemed potentially inappropriate for the elderly among family physicians' patients aged 65 and older (seniors) taking multiple prescribed medications. METHODS: Forty eight randomly selected family practices in 16 towns and cities in Southern Ontario, Canada and 889 of their senior patients were recruited into a randomized trial. We conducted a cross-sectional analysis of prescription insurance data from the provincial universal prescription insurance database over 12 months, from the 777 seniors who completed the trial and agreed to have their data released. The prevalence and patient and physician predictors of use of a potentially inappropriate medication (PIM), as defined by published widely accepted criteria, were examined. RESULTS: The median number of prescriptions filled was 24. Nearly one-fifth (16.3%) of the seniors received at least one prescription for a PIM, with short-acting benzodiazepine prescriptions for longer than 30 days (6.4%) and oxybutynin (3.7%) being the types prescribed most frequently. In univariate and multiple variable analyses, women were found to be statistically significantly more likely to be prescribed a PIM (adjusted OR = 1.6; 95% confidence interval = 1.0-2.4). Age, education, self-rated health, number of health conditions, and number of prescriptions were not associated with PIM use. Physician gender, family medicine certification status, and time since graduation were not significantly associated with PIM prescribing. CONCLUSIONS: Prescribing of PIMs, especially of short-acting benzodiazepines was common in seniors taking multiple medications. Interventions to reduce use of PIM, especially long-term benzodiazepines, are important in primary care. PMID- 15128684 TI - Academic detailing has no effect on prescribing of asthma medication in Danish general practice: a 3-year randomized controlled trial with 12-monthly follow ups. AB - BACKGROUND: Educational outreach visits, particularly when combined with social marketing, appear to be a promising approach to modifying health professional behaviour, especially prescribing. Results from previous studies have shown a varying effect. OBJECTIVE: The purpose of the study is to examine the effect of academic detailing as a method of implementing a clinical guideline in general practice. METHODS: A cluster randomized, controlled, blinded study was carried out of the effect of an academic detail visit compared with postal distribution of a guideline for prescribing asthma medication. Half the practices in a Danish county with 100 practices were visited once. The outcome measure was routinely collected data from all Danish pharmacies on the sales of asthma medication. Data were collected monthly for 2 years before to 1 year after the intervention. RESULTS: There was no effect on the pattern of prescription of asthma medicines following the visit, neither immediately nor long term. CONCLUSION: We found no effect of academic detailing as a single intervention. PMID- 15128685 TI - Experiences and understandings of social and emotional distress in the postnatal period among Bangladeshi women living in Tower Hamlets. AB - OBJECTIVE: The purpose of this study was to explore first-generation Bangladeshi women's understandings and experiences of postnatal distress, and to describe coping strategies during the postnatal period. METHODS: This was a qualitative study using focus groups. Subjects were drawn from three existing community groups in Tower Hamlets, a multiethnic, socially deprived borough in east London. Thematic content analysis was used to explore and present the data. RESULTS: Many women received little practical or emotional support once home from hospital with a new baby, because of the lack of extended family networks; this contrasts with the 40 day rest period common in Bangladesh. These women understood emotional distress as separate from physical symptoms or illness, and recognized that one may influence or cause the other. Distinctive language was used to describe these thoughts and feelings. The roles of health visitors, midwives and GPs were understood solely in terms of physical care. Accordingly, they did not access professionals for emotional or psychological problems. Lack of language support services contributed to the women not seeking help. CONCLUSIONS: Information about services, and professional roles in the postnatal period should be extended to include key family members such as husbands and mothers-in-law. Dialogue with Bangladeshi women may ensure that women understand the extended roles of GPs, health visitors and midwives in providing help for emotional distress, alongside their role in physical health care. More language support and advocacy is needed if women are to access the full range of health services. PMID- 15128686 TI - Use of information about maternal distress and negative life events to facilitate identification of psychosocial problems in children. AB - BACKGROUND: Despite the availability of effective screening measures, primary care physicians fail to identify and manage many children with psychosocial problems. Physicians often have information about significant negative events in a child's life. The present study evaluated the potential utility of using information about negative life events to facilitate physician identification of children with psychosocial problems. METHODS: Negative life events, maternal distress and child psychosocial functioning measures were completed by 185 mothers of children, aged 4-12 years. Family physicians provided data about the children's psychosocial functioning. RESULTS: Mothers identified 15.1% (n = 28) of the children as having psychosocial problems. Physicians correctly identified 21% (n = 6) of these at-risk children. Physician use of negative life events would have led to the identification of 39.2% (n = 11) at-risk children. Information about maternal distress and negative life events would have resulted in an additional 18% (n = 5) of children identified by the physicians. Information about maternal distress alone would have resulted in an identification rate of 53.5% (n = 15). CONCLUSIONS: Using information about negative events in a child's life, physicians could improve their rate of identification of children with psychosocial problems. Children who have had more than two negative events in their lives are at increased risk for psychosocial problems. PMID- 15128687 TI - Long-term benzodiazepine users in family practice: differences from short-term users in mental health, coping behaviour and psychological characteristics. AB - BACKGROUND: Contrary to short-term use, long-term benzodiazepine use is undesirable. Nevertheless, its prevalence is high. To prevent long-term use, it is important to know which short-term users are at risk of becoming long-term users. OBJECTIVES: The purpose of the present study was to identify patient related factors of long-term versus short-term use of benzodiazepines. METHODS: A cross-sectional study was carried out in family practices among users of benzodiazepines with regard to DSM-IV diagnosis, coping and psychosocial characteristics,. In a multivariate logistic regression analysis, long-term use of benzodiazepines was the dependent variable. RESULTS: A total of 164 short-term and 158 long-term benzodiazepine users participated in the study. Having a DSM-IV disorder and psychiatric co-morbidity, being older, less educated, lonely and using more avoidance coping behaviour was associated with long-term use of benzodiazepines compared with short-term use. CONCLUSION: The associations found point to possibilities to reduce long-term benzodiazepine use, for example if patients with these characteristics are treated with the alternatives to benzodiazepines or are monitored closely for a short period after being prescribing benzodiazepines. PMID- 15128688 TI - Implementation of brief alcohol interventions by nurses in primary care: do non clinical factors influence practice? AB - BACKGROUND: In the UK, GPs and practice nurses selectively provide brief alcohol interventions to risk drinkers. GPs' provision of a brief alcohol intervention can be predicted by patient characteristics, practitioner characteristics and structural factors such as the features of the practice and how it is organized. However, much less is known about possible modifiers of nurse practice. OBJECTIVE: Our aim was to investigate if patient characteristics, nurse characteristics and practice factors influence provision of a brief alcohol intervention by practice nurses in primary health care. METHODS: One hundred and twenty-eight practice nurses who had implemented a brief alcohol intervention programme in a previous trial based in the North of England were requested to screen adults presenting to their surgery and follow a structured protocol to give a brief intervention (5 min of advice plus an information booklet) to all 'risk' drinkers. Anonymized carbon copies of 5541 completed Alcohol Use Disorders Identification Test (AUDIT) screening questionnaires were collected after a 3 month implementation period and analysed by logistic regression analysis. RESULTS: Although AUDIT identified 1500 'risk' drinkers, only 926 (62%) received a brief intervention. Logistic regression modelling showed that patients' risk status as measured by AUDIT score was the most influential predictor of a brief intervention by practice nurses. However, risk drinkers who were most likely to receive a brief intervention were male. Patients' age or social class did not independently predict a brief intervention. The multilevel model was unable to identify any independent nurse characteristics that could predict a brief intervention, but indicated significant variation between nurses in their tendency to offer the intervention to patients. No structural factors were found to be positively associated with selective provision. CONCLUSIONS: Patient and nurse factors contributed to the selective provision of a brief intervention in primary care. If patients are to experience the beneficial effects of a brief alcohol intervention, then there is a need to improve the accuracy of delivery. PMID- 15128689 TI - The role of GPs in treating psychological distress: a study of midlife Australian women. AB - BACKGROUND: Patient satisfaction with general practice care is important for treatment adherence, yet little is known about women's satisfaction with general practice care in relation to emotional problems. OBJECTIVES: The purpose of the present study was to explore women's perceptions of the help provided by GPs for psychological distress. METHODS: Qualitative and quantitative data were gathered using semi-structured telephone interviews in NSW Australia. The respondents were 322 women aged 45-50 who participated in the baseline survey of Women's Health Australia (WHA). RESULTS: Of the 309 women who had had a period of distress in the previous 12 months, 159 [52%, confidence interval (CI) 46.4-57.6] had talked to a GP about their difficulties. Listening was the main help given by GPs (68%, CI 60.7-75.3), followed by a prescription for medication (55%, CI 47.2-62.8) and referral to specialist care (13%, CI 7.8-18.2). Few women reported specific behavioural interventions, such as counselling (4%, CI 0.9-7.1) or relaxation (1%, CI 0 to 2.6). There was a relatively high degree of satisfaction with referral, counselling and relaxation advice amongst those who received these treatments. In contrast, a fifth of women who received a prescription or were listened to found these treatments unhelpful (20%, CI 11.6-28.4; and 21%, CI 14.2 29.8, respectively). Thematic analysis highlighted three main concerns for women, namely structural limitations of the GP-patient consultation, GPs' limited interpersonal skills and GPs' limited interest, knowledge and skills in mental health. CONCLUSION: While most women find their general practic care helpful, many reported shortcomings in terms of both GP skills and structural limitations of the consultation. These findings are useful in informing the development of training programmes for GPs. PMID- 15128690 TI - The incidence and characteristics of end-of-life decisions by GPs in Belgium. AB - BACKGROUND: Medical end-of-life decisions (ELDs) have been discussed for several years in different countries, but little is known about the involvement of GPs in these ELDs. OBJECTIVES: The aim of the present study was to establish the incidence and characteristics of ELDs by GPs. METHOD: We selected 3999 deaths, a 20% random sample of all registered deaths during the first 4 months of 1998 in Flanders, Belgium, and mailed anonymous questionnaires to the attesting physicians. Here we focus exclusively on the 1647 deaths certified by GPs. RESULTS: The GPs returned 1067 questionnaires (response rate of 64.8%). At least one ELD was made in 39.5% [95% confidence interval (CI) 37.8-41.2] of all primary care deaths. The incidence of euthanasia (including physician-assisted suicide) was 1.5% (95% CI 0.9-2.3) (incidence higher among more educated patients and at home), of administration of lethal drugs without the patient's explicit request 3.8% (95% CI 2.9-5.0) (higher among cancer patients), of alleviation of pain and symptoms with possibly life-shortening effect 18.6% (95% CI 17.0-20.2) (higher among cancer patients and married patients) and of non-treatment decisions 15.6% (95% CI 14.2-17.2) (higher among cancer patients and in nursing homes). The decision was not discussed with the patient in three out of four of the ELDs. A colleague was consulted in one in four ELD cases. CONCLUSION: ELDs are common in general practice in Flanders, Belgium, despite the restrictive law concerning euthanasia at the time of this study. The incidence of these ELDs varies with cause and place of death, the patient's education and the GP's religion and age. Requirements of prudent practice regarding ELDs are rather poorly met by GPs. Further international research and debate is needed to highlight the GPs' important role in end-of-life care. PMID- 15128691 TI - How research-conscious GPs make decisions about anticoagulation in patients with atrial fibrillation: a qualitative study. AB - BACKGROUND: Delays in the implementation of research findings have been of particular concern during the last decade. New findings, such as the use of warfarin to prevent stroke in patients with atrial fibrillation, have been found to be implemented patchily in clinical practice, in both primary and secondary care. OBJECTIVE: The purpose [corrected] of the study was to explore how GPs with an active interest in research or evidence-based medicine (EBM) make decisions about anticoagulation in patients with atrial fibrillation. METHODS: Semi structured interviews with GPs about their experiences in managing patients with atrial fibrillation were recorded on audio-tape, transcribed and analysed using the 'Framework' method. A constructivist approach was taken to analysis and interpretation. RESULTS: Eleven interviews were included in the analysis. Two key themes, 'evidence' and 'professional role', were identified. No two respondents had the same perception of the evidence, which was influenced by experience, attitudes and a variable knowledge of the literature. Recent publications about the effectiveness of aspirin compared with warfarin, and the publication Clinical Evidence were the most frequently mentioned sources of evidence. GPs with confidence in EBM skills described giving highly detailed explanations to patients and having a great commitment to shared decision making, even if this resulted in patients declining treatment. For this reason, they also expressed antagonism towards prescriptive clinical guidelines. Hospital doctors were seen as exerting a powerful influence on decisions, as being 'disease-centred', difficult to challenge and poor at communicating. CONCLUSIONS: Decision making about anticoagulation is complex and is determined by a socially constructed view of the evidence strongly influenced by the GP's professional role. PMID- 15128692 TI - Less haste more speed: factors that prolong the interval from presentation to diagnosis in some cancers. AB - BACKGROUND: In the UK, the GP is the gatekeeper to specialist services in addition to many other roles. Recently, the GP is also expected to select cases that warrant 'urgent' as opposed to 'routine' specialist investigation. Failure to refer on the appropriate timetable may have implications for timely diagnosis. OBJECTIVE: Our aim was to explore the circumstances in which the diagnosis of cancer is delayed with reference to the primary care records and by a structured investigation of clinical records in one practice. METHODS: The study was set in an urban group practice serving a mixed population of deprived and affluent communities. List size was 10 440 patients, with five whole time equivalent partners and three practice nurses. The appointment system was fully computerized and there were no personal lists. Records for all cases with specified common cancers diagnosed since 1990 and still registered in the practice were reviewed. The interval from presentation to referral, referral to diagnosis and presentation to diagnosis was compared for a series of factors including 'urgent' referral. The clinical team currently working in the practice conducted a structured review of the case records for the most delayed cases. RESULTS: Fifty four cases were listed in the practice. A series of factors were identified as having a bearing on delayed diagnosis, including a reticence on the part of patients to seek to expedite specialist appointments, failures of communication, and patients presenting multiple problems in short general practice consultations. The action plan agreed by the clinical team includes improving the quality of communication with secondary care, follow-up of patients who have been referred for radiological or ultrasound investigation and reviewing patients who fail to attend specialist clinics. CONCLUSIONS: The data imply that delays sometimes result from avoidable errors before and after referral and especially by the patient entering secondary care on the wrong pathway. Improving the patients' experience in health care requires the provider to take a global view of the service. Primary care is not merely a filter but influences and is influenced by policies in other parts of the health care system. PMID- 15128693 TI - Secondary prevention of coronary heart disease is disappointing among patients of working age. AB - BACKGROUND: The risk factors underlying coronary heart disease (CHD) are well known. OBJECTIVE: The purpose of this study was to evaluate risk factors related to secondary prevention of working-age CHD patients. METHODS: CHD patients with (n = 139) and without (n = 203) myocardial infarction were selected from a postal questionnaire study (n = 21 101) of randomly selected Finns aged 20-54 years (HeSSup study). Four age- and sex-matched controls were chosen for every patient. RESULTS: CHD patients still smoke, are obese and suffer hangovers more frequently than the control population. CONCLUSION: The health care system has not succeeded in the secondary prevention of CHD. PMID- 15128694 TI - Current practice and future interest of GPs and prospective parents in pre conception care in The Netherlands. AB - BACKGROUND: Evidence for the effectiveness of pre-conception care is growing. GPs are among the possible providers of pre-conception care. OBJECTIVES: Our first aim was to investigate current pre-conception care activities of GPs and to determine whether prospective parents would visit the GP in the existing health care system. A further aim was to determine the interest of GPs and prospective parents in the introduction of pre-conception care clinics. METHODS: A survey was conducted among 200 GPs and 303 recently married couples (606 individuals). RESULTS: Of the eligible GPs, 52% (n = 102) participated, as did 70% (n = 381) of the eligible individuals who are planning a pregnancy. In the current situation, only a few individuals visit their GP pre-conceptionally, and many GPs do not discuss family history or consanguinity with prospective parents. In general, the GPs and prospective parents had a positive attitude towards the introduction of pre-conception care clinics. CONCLUSION: Since most GPs and prospective parents favoured the introduction of pre-conception care, future research should focus on the most appropriate way of implementation. PMID- 15128695 TI - The needs of patients dying of chronic obstructive pulmonary disease in the community. AB - BACKGROUND: Ninety percent of the last year of life of a terminally ill patient is spent at home, but possible drawbacks to domicillary terminal care have been suggested. Most previous studies have taken place in secondary care settings, and have often obtained relatives' views of patients' needs rather than those of the patient. OBJECTIVES: Our aim was to determine the needs of patients dying in primary care from chronic obstructive pulmonary disease (COPD). METHODS: Semi structured interviews were undertaken with 16 patients who were having maximal therapy for COPD, who were thought likely to die in the year following the commencement of the study. RESULTS: Five themes were identified. (i) Patients' information needs and the future course of their illnesses. Information needs were often variable and patients were sometimes unwilling to contemplate the future. (ii) The impact of symptoms on patients' lives. Dyspnoea was a particular problem. (iii) Attitudes towards and opinions on smoking, which many acknowledged as being causal. (iv) Attitudes to help received. Many patients relied on their family; most patients felt their doctors were helpful although some expressed reservations. (v) Patients' expressed needs. These usually focused on mobility, and many patients would not express needs even when prompted. CONCLUSION: Poor symptom control remains an important cause of distress. The low number of expressed needs may reflect patients' unwillingness to appear ungrateful, but the variability of information needs emphasizes the importance of an individual approach to patients with an apparently homogenous disease PMID- 15128696 TI - Duration of symptoms and follow-up patterns of patients discharged from the emergency department after presenting with abdominal or flank pain. AB - BACKGROUND: Little is known about the duration of symptoms and follow-up patterns of patients seen in emergency departments for abdominal or flank pain. OBJECTIVE: We aimed to measure the duration of symptoms and follow-up rate of patients discharged home from the emergency department after presenting with non-traumatic abdominal or flank pain. METHODS: We conducted a single-centre, prospective descriptive study of adult patients who presented to our emergency department with non-traumatic abdominal or flank pain and were discharged from the emergency department. We gathered clinical data during the index emergency department visit and conducted telephone interviews of subjects 2-5 weeks later. RESULTS: We reached 63 of 90 subjects (70%). The median duration of pain was 3 days after the emergency department visit. During the follow-up period, only 41% had followed-up with their family physician or primary care provider, although an additional 21% had planned to. Persistence of symptoms was common in the 37% of subjects who did not follow-up. CONCLUSION: Of subjects discharged from the emergency department after visits for non-traumatic abdominal or flank pain, most improve within several days. Fewer than half follow-up with a family practitioner or a primary care provider. Emergency department revisits are uncommon and often for unrelated problems. PMID- 15128697 TI - Acute respiratory symptoms in adults in general practice. AB - BACKGROUND: Community studies have shown that approximately 30% of patients with acute respiratory tract symptoms have no identifiable infective aetiology. This may not be applicable in general practice. OBJECTIVE: The purpose of this study was to determine the infective aetiology in patients who presented to primary care doctors with acute respiratory symptoms. METHODS: A prospective study was carried out in all nine primary care clinics belonging to the National Healthcare Group Polyclinics (NHGPs) in Singapore. The subjects comprised 594 consecutive patients (318 males, 276 females) aged > or = 21 years who presented with complaints of any one of cough, nasal or throat symptoms of <7 days duration. Data collection was through interview using structured questionnaire, physical examination, throat swabs for bacterial culture and nasal swabs for virus identification by immunofluorescence (IF) and polymerase chain reaction (PCR). Additional PCR was performed on a subsample of 100 patients. Patients were followed-up until resolution of symptoms. RESULTS: The aetiological diagnosis by infective agent is as follows: 150 patients (25.2%) had virus infections, of which 90.7% (136/150) were by rhinovirus. Fourteen patients (2.4%) had bacterial infections, of which 10 were due to group G streptococcus. Group A streptococcus was not detected. Nineteen patients with new pathogens were identified by further PCR. These included parainfluenza 4, human coronavirus OC43, adenovirus, enterovirus and Chlamydia pneumoniae. No pathogen could be identified in 49% of patients. There were no differences in clinical presentation and socio demographic variables between patients who had viral infections and those in whom no pathogen could be identified. CONCLUSION: In about half of patients who presented at NHGPs, no pathogens could be identified even after PCR. A non infective aetiology could be considered in these patients. PMID- 15128698 TI - Selections from current literature. The spectrum of treatment options for obesity. PMID- 15128699 TI - Internet-based surveys of health professionals. PMID- 15128700 TI - Over-expression of angiotensin converting enzyme-1 augments cardiac hypertrophy in transgenic rats. AB - Increased cardiac angiotensin converting enzyme-1 (ACE1) is found in individuals who carry a deletion in intron 16 of ACE1 gene or in individuals who suffer from cardiac disorders, such as hypertrophy. However, whether a single increase in ACE1 expression leads to spontaneous cardiac defects remains unknown. To determine if the increased cardiac ACE1 actively plays a role or is merely the consequence of pathological changes in the process of cardiac hypertrophy, we generated a transgenic rat model with selective over-expression of human ACE1 in the cardiac ventricles. The left ventricular ACE1 activity is elevated about 50 fold in transgenic rats. Angiotensin-1 perfusion of isolated hearts demonstrated a significant decrease in coronary artery flow compared with non-transgenic littermates, suggesting that the transgenic ACE1 is functional. Neither cardiac hypertrophy nor other morphological abnormalities were observed in transgenic rats under standard living conditions. It was found, however, after induction of hypertension by suprarenal aortic banding, that the degree of cardiac hypertrophy in transgenic rats was significantly higher than that of banded control rats. The expressions of both ANF and collagen III, molecular markers of cardiac hypertrophy, were also increased in banded transgenic rats compared with banded control. Our results suggest that increased cardiac ACE1 does not trigger but augments cardiac hypertrophy. PMID- 15128701 TI - Corticosteroid pharmacogenetics: association of sequence variants in CRHR1 with improved lung function in asthmatics treated with inhaled corticosteroids. AB - Corticosteroids mediate a variety of immunological actions and are commonly utilized in the treatment of a wide range of diseases. Unfortunately, therapy with this class of medications is associated with a large proportion of non responders and significant side effects. Inhaled corticosteroids are the most commonly used asthma controller therapy. However, asthmatic response to corticosteroids also varies widely between individuals. We investigated the genetic contribution to the variation in response to inhaled corticosteroid therapy in asthma. The association of longitudinal change in lung function and single nucleotide polymorphisms from candidate genes crucial to the biologic actions of corticosteroids were evaluated in three independent asthmatic clinical trial populations utilizing inhaled corticosteroids as the primary therapy in at least one treatment arm. Variation in one gene, corticotropin-releasing hormone receptor 1 (CRHR1) was consistently associated with enhanced response to therapy in each of our three populations. Individuals homozygous for the variants of interest manifested a doubling to quadrupling of the lung function response to corticosteroids compared with lack of the variants (P-values ranging from 0.006 to 0.025 for our three asthmatic populations). As the primary receptor mediating the release of adrenocorticotropic hormone, which regulates endogenous cortisol levels, CRHR1 plays a pivotal, pleiotropic role in steroid biology. These data indicate that genetic variants in CRHR1 have pharmacogenetic effects influencing asthmatic response to corticosteroids, provide a rationale for predicting therapeutic response in asthma and other corticosteroid-treated diseases, and suggests this gene pathway as a potential novel therapeutic target. PMID- 15128702 TI - Fxr1 knockout mice show a striated muscle phenotype: implications for Fxr1p function in vivo. AB - FXR1 is one of the two known homologues of FMR1. FXR1 shares a high degree of sequence homology with FMR1 and also encodes two KH domains and an RGG domain, conferring RNA-binding capabilities. In comparison with FMRP, very little is known about the function of FXR1P in vivo. Mouse knockout (KO) models exist for both Fmr1 and Fxr2. To study the function of Fxr1 in vivo, we generated an Fxr1 KO mouse model. Homozygous Fxr1 KO neonates die shortly after birth most likely due to cardiac or respiratory failure. Histochemical analyses carried out on both skeletal and cardiac muscles show a disruption of cellular architecture and structure in E19 Fxr1 neonates compared with wild-type (WT) littermates. In WT E19 skeletal and cardiac muscles, Fxr1p is localized to the costameric regions within the muscles. In E19 Fxr1 KO littermates, in addition to the absence of Fxr1p, costameric proteins vinculin, dystrophin and alpha-actinin were found to be delocalized. A second mouse model (Fxr1 + neo), which expresses strongly reduced levels of Fxr1p relative to WT littermates, does not display the neonatal lethal phenotype seen in the Fxr1 KOs but does display a strongly reduced limb musculature and has a reduced life span of approximately 18 weeks. The results presented here point towards a role for Fxr1p in muscle mRNA transport/translation control similar to that seen for Fmrp in neuronal cells. PMID- 15128703 TI - Partial loss of presenilins causes seborrheic keratosis and autoimmune disease in mice. AB - Presenilin (PS1) and (PS2) are the centers of gamma-secretase that release Abeta from APP in Alzheimer's disease (AD). They cleave signaling proteins like Notch and downregulate beta-catenin to modulate Wnt signaling. Inactivation of PS1 or PS1 and PS2 causes a prenatally lethal 'Notch phenotype,' which has hampered investigation of PS function in adulthood seriously. We have thus turned towards PS1+/-PS2-/- mice which carry the most severe reduction of PS alleles compatible with survival, to analyze the consequences of impaired PS function especially in adulthood. In these 'partial deficient' mice, PS1 protein concentration is considerably lowered, functionally reflected by reduced gamma-secretase activity and impaired beta-catenin downregulation. Their phenotype is normal up to approximately 6 months, when the majority of the mice develop an autoimmune disease characterized by dermatitis, glomerulonephritis, keratitis and vasculitis, as seen in human systemic lupus erythematosus. Besides B-cell dominated infiltrates, we observe a hypergammaglobulinemia with immune complex deposits in several tissues, high-titer nuclear autoantibodies and an increased CD4+/CD8+ ratio. The mice further develop a benign skin hyperplasia similar to human seborrheic keratosis as opposed to malignant keratocarcinomata observed in skin-specific PS1 'full' knockouts. A partial reduction of PS function in PS1+/ PS2-/- mice causes a novel phenotype in adulthood unrelated to the developmental defects of full knockouts. As PS1+/-PS2+/- mice remain healthy, this points towards a sharply defined minimum of PS function. Skin and immune system appear to be especially sensitive targets of impaired PS function and may need careful monitoring if gamma-secretase inhibitors are envisaged for treating AD. PMID- 15128704 TI - Molecular pathogenesis of cystinosis: effect of CTNS mutations on the transport activity and subcellular localization of cystinosin. AB - Cystinosis is an inherited disorder characterized by defective lysosomal efflux of cystine. Three clinical forms (infantile, juvenile and ocular cystinosis) have been described according to the age of onset and severity of the symptoms. The causative gene, CTNS, encodes a seven transmembrane domain protein, cystinosin, which we recently identified as a H+-driven cystine transporter using an in vitro transport assay. In this study, we explored the relationship between transport activity and intracellular localization of cystinosin mutants and their associated clinical phenotype. Thirty-one pathogenic mutations (24 missense mutations, seven in-frame deletions or insertions) were analysed. Most of the mutations did not alter the lysosomal localization of cystinosin, although three partially mislocalized the protein independently of its C-terminal sorting motif, thus confirming the presence of an additional sorting mechanism. Sixteen of 19 mutations associated with infantile cystinosis abolished transport, whereas three of five mutations associated with juvenile or ocular forms strongly reduced transport, in agreement with the milder clinical phenotype. Five atypical, unclassified or misclassified mutations could be clarified using the transport data and additional genetic information. Overall, our data demonstrate that, excluding premature termination of cystinosin, impaired transport is the most frequent cause of pathogenicity, with infantile cystinosis generally resulting from a total loss of activity. Thus the transport assay could be used as a prognostic tool when novel mutations are identified. PMID- 15128705 TI - Getting evidence into policy and practice to address health inequalities. PMID- 15128706 TI - Effect evaluation of a comprehensive community intervention aimed at reducing socioeconomic health inequalities in The Netherlands. AB - To date, comprehensive community health projects have not been evaluated in terms of their effect at the individual level, because outcomes are usually not defined at this level. In a community health project in The Netherlands, evaluation outcome mapping, a technique derived from intervention mapping, was used to identify distal as well as proximal programme objectives from which outcome measures could be derived. The intervention took place in a deprived area, where community members themselves defined stress, lack of area safety and parenting problems as the health-related problems they wanted to see addressed in the project. Local organizations wrote and implemented an action plan. The effects among residents were studied in a quasi-experimental design. Although no significant effects on improved perceived health or health-related problems were found at the level of the residents, the problems identified and their assumed causes showed significant coherence. This study is believed to be of relevance to evidence-based health promotion theory and practice as it demonstrates that it is possible to conduct an individual effect evaluation in a comprehensive community approach without jeopardizing the process of the intervention. PMID- 15128707 TI - Health promotion for socially disadvantaged groups: the case of homeless older men in Australia. AB - There is extensive evidence that health promotion routinely benefits those who are already most socioeconomically advantaged. While the government's healthy ageing policy recognizes that improving health outcomes will require a range of strategies involving different target groups, recommendations focus on the issues and needs of the comfortable majority. This paper examines the scope and relevance of health promotion for one disadvantaged minority with extensive health needs: homeless older men. In an ethnographic study of older men (> or = 50 years of age) living alone in the inner city (Sydney), 32 men were identified as homeless and are the focus of this paper. Face to face semi-structured interviews were used to record the men's accounts of their everyday lives, including their health and use of services. The conditions in which these men were living were observed and recorded, and the researchers were aware of health and other services available in the geographic area. All informants were living on or below the poverty line. They reported a range of health conditions, for which many accessed available mainstream and specialist health services. Some obstacles to accessing services were noted. Information relevant to widely endorsed prescriptions for 'healthy ageing' also emerged. These included physical activity (especially walking), healthy eating, social activity and adopting healthy lifestyle habits. Findings highlight the extent to which these men lack the basic requirements for healthy ageing, notably adequate incomes and housing. At the same time, within the constraints of the lifestyle they lead, they are motivated to maintain their health and independence. While there are limits to what can be achieved for such people at a local level of service delivery, it is possible to identify feasible health promotion goals and service strategies. PMID- 15128708 TI - ask for it: development of a health advocacy intervention for adults with intellectual disability and their general practitioners. AB - Two per cent of people in Australia have intellectual disability and the adults in this population often have poor health status. This poor health can be partly attributed to communication difficulties encountered by people with intellectual disability and also health professionals in consultation settings. The design and development processes of an educational intervention to improve communication between patients, general practitioners (GPs) and also advocates in a population of adults with intellectual disability are described. The design process was collaborative and involved adults with intellectual disability, GPs, parents, support workers and other professionals. It was a nine-step development process and led to the final communication tool package, the ask (advocacy skills kit) 5 year health diary and educational session. As a result of the collaborative design and development processes, this diary included qualities not found in most other medical record keeping systems: visual appeal, advice on how to be a health advocate, utility for a range of users, privacy, portability and sufficient capacity to record personal patient information which enhanced communication between doctor, patient and advocate. It is proving to be very popular. Clear implications were found for applying established criteria and incorporating the needs of users in the design of educational interventions in the intellectually disabled population. Health promotion tools aiming to improve the current poor health status of adults with intellectual disability should be developed further. PMID- 15128709 TI - Wide range of socioeconomic factors associated with mortality among cities in Japan. AB - The aim of this study was to identify socioeconomic factors associated with mortality among cities in Japan. Sex-specific and age-adjusted mortality rates for 1990 and 1995 were calculated by 779 local administrative units across the nation. One hundred indicators related to socioeconomic factors were compiled and divided into eight categories: economy, education, living conditions, vegetation and open space, transport, preventive activities, medical care and demography. Composite socioeconomic indices were formulated using factor analysis of the socioeconomic indicators by category, and the association between the indices and mortality rates was examined by correlation analysis and multiple regression analysis. Nineteen composite socioeconomic indices were obtained from factor analysis, and all indices except educational expenditure-related index were significantly correlated with mortality rates. Unemployment, old housing, primary health resources and density were independently positively associated, and higher education, public library activity, health check-up participation and population growth were independently negatively associated with both 1990 and 1995 male mortality rates. For female mortality, higher income, unemployment, spacious dwelling, old housing, less vegetation, road facility, numbers of cars per population, primary health resources and density were independently positively associated, and higher education, public library activity and health check-up participation were independently negatively associated. The relationship between mortality and socioeconomic conditions was stronger in males than in females, and higher income and less vegetation were associated with higher mortality only for females. The present study demonstrated a close link between mortality and a wide range of socioeconomic conditions by using a number of indicators compiled from various data sources. The results promote a deeper understanding of socioeconomic health determinants and development of multi-sectoral health policy to improve population health. PMID- 15128710 TI - A comparison of smoking behaviors among medical and other college students in China. AB - A survey of students' smoking in China (n = 1896), comparing medical students with college students in non-medical majors, was carried out to determine whether a medical education has a preventive effect on smoking uptake. The survey, sampling students from 12 universities in three cities, found no significant differences between medical and non-medical students in smoking prevalence (40.7% versus 45.1% for males, 4.4% versus 6.0% for females), in 'ever smoked' groups, in 'ever smoked 100 cigarettes' groups or in years of smoking. For both student groups, smoking prevalence increased with age and with years of college. However, one significant difference was found among the smokers: medical students were more likely to be occasional smokers than were non-medical students (75.3% occasional smokers among medical students who smoked versus 60.6% among non medical students). These results suggest that a medical education had little effect on these students' decisions to smoke, but that it may have modified their consumption level. Future studies are needed to ascertain factors affecting the decision to smoke and to identify possible early adopters of a non-smoking culture in China. Action on a societal level is urgently needed to change Chinese social norms regarding smoking. PMID- 15128711 TI - Does the Heartbeat Award scheme in England result in change in dietary behaviour in the workplace? AB - The Heartbeat Award (HBA) scheme is a national nutrition labelling scheme that operates throughout England. The aim of this study was to assess whether the implementation of the HBA scheme in the workplace results in an improvement in eating habits at work. A longitudinal survey of employees using a structured questionnaire pre- and post-HBA intervention in six workplaces in Leicestershire, England, was conducted. A qualitative food frequency questionnaire was used to assess dietary behaviour 6 months before the scheme was implemented and 6 months afterwards. Four HBA workplaces (n = 453 employees) were compared with two comparison workplaces (n = 124 employees). The outcome measures for dietary change were consumption of 20 food items, i.e. 16 food-frequency and four food type items. Changes in pre-intervention data compared with post-intervention data evaluated the impact of the HBA on eating habits. Differences were considered statistically different at the p < 0.05 level. Crude and adjusted odds ratios were used to assess differences in change in dietary behaviour between the intervention and comparison workplaces. The results showed that there was significantly more positive change in intervention workplaces only (i.e. the changes were not detected in the comparison workplaces) for four of the 20 food items tested: increase in consumption of fruit (p = 0.029); reduction in consumption of fried foods (p = 0.044) and sweet puddings (p = 0.042); and change to lower fat milks (p = 0.034). In conclusion, the HBA had a modest impact on dietary intake, resulting in a significant positive change in four out of 20 foods consumed by employees in HBA-holding premises. PMID- 15128712 TI - The Healthy Bus project in Denmark: need for an action potential assessment. AB - Research over the last 50 years has repeatedly documented that bus drivers are exposed to several physical and psychological risk factors, which are associated with health problems in the form of heart, musculo-skeletal and stomach disease, and increased coronary mortality. So why has there been little action to improve the situation when it is so obviously indicated by such assessments? This article describes the long and complex process that has made it possible to launch almost 200 interventions among the 3500 municipal bus drivers in Copenhagen. Using a participative action research design, new evidence was gathered by broadening the traditional work environmental scope to lifestyle, health issues and private matters. Comparing this updated needs assessment with a national reference population, it was found that drivers were often still worse off. Again, simply presenting new evidence did not seem to lead to changes and further work is needed to empower the stakeholders so that they can commit to start making effective interventions. It is concluded that every needs assessment has to be supplemented with an evaluation of the action potential. PMID- 15128713 TI - A study of safety-promoting activities for children and adolescents in 25 Swedish municipalities. AB - The purpose of this study was to investigate two elements of the community approach to safety promotion for children and adolescents. The first concerns whether there is any correlation between safety-promoting activities in different municipal settings, and the second whether socio-economic and demographic characteristics affect the level of safety-promoting activities. A cross sectional study was performed comprising 25 municipalities in Stockholm County. The extent of safety-promoting activities was studied in four settings: the municipal setting in general, the traffic setting, the pre-school setting, and the school and leisure activity setting. Data on nine demographic and socio economic characteristics of the municipalities were collected from public statistics. There were three main findings. First, safety-promoting activities in the municipal setting in general, the pre-school setting, and the school and leisure activity setting, but not in the traffic setting, were apparently associated with total municipal safety-promoting activities. Secondly, socio economic characteristics, measured as the proportion of adults with >12 years of education, did not predict the extent of safety-promoting activities. Finally, growing municipalities with a large proportion of children reported fewer safety promoting activities than non-growing municipalities. This study demonstrated weak correlations between safety-promoting activities in different municipal settings. The finding that growing municipalities have fewer safety-promoting activities provides pertinent lessons for future action and research. The study offers a method by which it may be possible to better assess the complex components of the community approach to safety promotion. PMID- 15128714 TI - A review of school drug policies and their impact on youth substance use. AB - Youth substance use is an important social and health problem in the United States, Australia and other Western nations. Schools are recognized as important sites for prevention efforts and school substance use policies are a key component of health promotion in schools. The first part of this paper reviews the known status of school policies on tobacco, alcohol and other illicit drugs in a number of Western countries and the existing evidence for the effectiveness of school drug policy in preventing drug use. The review shows that most schools in developed countries have substance use policies but that there is substantial variation in the comprehensiveness of these policies (i.e. the breadth of people, places and times of day that are explicitly subject to policy prohibitions), and the orientation of their enforcement (e.g. punitive versus remedial), both across and within schools. The few studies of policy impact focus solely on tobacco policy and provide preliminary evidence that more comprehensive and strictly enforced school policies are associated with less smoking. The second part of the paper introduces the International Youth Development Study, a new longitudinal research project aimed at comparing school policies and the developmental course of youth drug use in the United States, where drug policies are abstinence-based, with Australia, which has adopted a harm minimization approach to drug policy. PMID- 15128715 TI - Behavior change intervention research in community settings: how generalizable are the results? AB - This review examines the extent to which recent behavioral intervention studies conducted in community settings reported on elements of internal and external validity, with an emphasis on whether research has been conducted in representative settings with representative populations. A targeted review was conducted on community-based intervention studies that promoted good nutrition, physical activity or smoking cessation/prevention, and were published in 11 leading health behavior journals between 1996 and 2000. The RE-AIM framework (reach, efficacy, adoption, implementation and maintenance) was used to evaluate the extent to which each paper reported on elements of reach, efficacy/effectiveness, adoption, implementation and maintenance. A total of 27 publications were reviewed. Although most studies (88%) reported participation rates among eligible members of the target audience ('reach'), only 11% of studies reported the participation rate ('adoption') among eligible community based organizations or settings. Few studies reported if participating individuals or settings were representative of those found in the broader population. Although a majority of studies (59%) reported whether the intervention was delivered ('implementation'), few reported whether individuals maintained behavior change (30%) or whether organizations maintained or institutionalized interventions (0%). To increase the potential to translate community research findings to practice, studies should place a greater emphasis on obtaining and reporting external validity information, such as representativeness. The lack of external validity information limits researchers' and practitioners' ability to judge the generalizability of effects and the comparative utility of interventions. Improved reporting will facilitate implementation of proven and broadly applicable intervention strategies in communities. To make significant progress, all parties, including researchers, reviewers, editors and funders, need to take responsibility for increased emphasis on external validity information and ask what role they can best play to facilitate this process. PMID- 15128716 TI - Advocating evidence-based health promotion: reflections and a way forward. AB - In the past few years, significant advances have been made in health promotion to generate readily accessible systematic reviews of evidence on the effectiveness of interventions and programs. The influence of this evidence on policy and practice has, however, been unpredictable, and proponents of evidence-based practice are identifying ways to increase the use of research in decisions about health promotion interventions. This paper examines the following questions: (i) is the evidence that is available on the effectiveness of interventions actually relevant and useful to current policy and practice contexts?; and (ii) what is the researcher's or reviewer's role in interpreting the available evidence and advocating action based on their interpretations? The paper concludes by proposing an 'evidence-agenda map' to assist advocates of evidence-based policy and practice to identify the health promotion goals they seek to influence against the required and available evidence. PMID- 15128717 TI - The European Health Promoting Hospitals (HPH) project: how far on? AB - Back in 1986, the World Health Organization (WHO) produced the Ottawa Charter for Health Promotion. The intention of the charter was to create a framework that conveyed the notions of capacity building into a structured process for health promotion action in specific settings. This charter subsequently provided the vehicle from which the Health Promoting Hospital (HPH) initiative was launched, culminating in the Budapest Declaration of Health Promoting Hospitals (WHO, 1991). The aim of this paper is to investigate the nature and progress of the European HPH movement. Despite the fact that 'pockets' of concerted and progressive activity and evaluation have emerged from the HPH initiative, the majority of the available literature demonstrates a more limited impact than perhaps the WHO might have anticipated for its efforts over the last 15 years or so. Indications are that many of the member European HPH states have struggled to move beyond the 'project' phases of their planned programmes. This is not to detract from the considerable efforts that have been made to establish HPH networks or the continuing attempts to recruit further members/institutions into the movement. Nevertheless, this account concludes that a more concerted evaluation of European HPH progress is needed to accurately measure its impact and progress. If the situation remains unchanged, perhaps a fundamental review of the strategy is worth considering. PMID- 15128718 TI - Toronto charter outlines future health policy directions for Canada and elsewhere. PMID- 15128720 TI - Dietary sodium and blood pressure. PMID- 15128721 TI - Molecular characterization of class 1 integrons from Irish thermophilic Campylobacter spp. AB - OBJECTIVES: In this study a large random collection (n = 378) of Irish thermophilic Campylobacter isolates were investigated for the presence of integrons, genetic elements associated with the dissemination of antimicrobial resistance. METHODS: Purified genomic DNA from each isolate was analysed by PCR for the presence of class 1 integrons. Four gene cassette-associated amplicons were completely characterized. RESULTS: Sixty-two of the isolates possessed a complete class 1 integron with a recombined gene cassette located within a 1.0 kb amplicon containing an aadA2 gene. This cassette was present in both Campylobacter jejuni and Campylobacter coli isolates and following sequence analysis was shown to be similar to sequences recently reported in Salmonella enterica Hadar and on an 85 kb plasmid conferring quinolone resistance in Escherichia coli. CONCLUSIONS: Aminoglycoside aadA2-encoding class 1 integrons were identified among unrelated Campylobacter spp. Amino acid sequence comparisons revealed identical structures in both Salmonella and E. coli. The presence of class 1 integrons in Campylobacter spp. may be significant should these organisms enter the food chain and especially when antimicrobial treatment for severe infections is being considered. PMID- 15128722 TI - Antibiotic resistance in 3113 blood isolates of Staphylococcus aureus in 40 Spanish hospitals participating in the European Antimicrobial Resistance Surveillance System (2000-2002). AB - OBJECTIVES: Since 1998 the European Commission has funded EARSS. We present the antibiotic susceptibility results of invasive Staphylococcus aureus obtained in Spain (2000-2002). MATERIAL AND METHODS: Forty hospitals participated in this study, covering nearly 30% of the Spanish population. All blood isolates of S. aureus were included. Laboratories used their usual methods to perform microbiological studies. Annual external quality controls were carried out. A questionnaire with hospital, patient and specimen data was completed for each isolate. Results were included in a database and analysed with WHONET 5 software. RESULTS: Invasive S. aureus was isolated in 3113 patients. Resistance was 24.5% to oxacillin, 25.4% to ciprofloxacin, 25.2% to erythromycin and 12.1% to gentamicin. Gentamicin resistance decreased from 16.6% (2000) to 9.7% (2002). Multiresistance was observed in 68.1% of oxacillin-resistant isolates. More prevalent multiresistance profiles consisted of oxacillin-ciprofloxacin erythromycin-gentamicin (7.4%) and oxacillin-ciprofloxacin-erythromycin (7.1%). Oxacillin resistance was significantly higher in nosocomial isolates than in those implicated in community-onset infections (26.7% versus 14.2%), in isolates from adults than in those from children (27.3% versus 4.7%), in hospitals with >500 beds than in those with <500 beds (31.1% versus 18.3%) and in isolates from Intensive Care Units than in those from other departments (39.3% versus 24%). Decreased susceptibility to vancomycin was not detected. CONCLUSIONS: In Spain, S. aureus blood isolates present a high prevalence of resistance to oxacillin, ciprofloxacin and erythromycin, as well as a high prevalence of multiresistance. Oxacillin resistance remains stable but varies in relation to hospital size, patient age, hospital department and place of infection acquisition. PMID- 15128723 TI - Antimicrobial susceptibility of the pathogens of bacteraemia in the UK and Ireland 2001-2002: the BSAC Bacteraemia Resistance Surveillance Programme. AB - OBJECTIVES: To describe the current patterns of antimicrobial resistance in the major pathogens of bacteraemia in the UK and Ireland, to highlight any unexpected resistance patterns and to act as a reference baseline for future studies. METHODS: In 2001 and 2002, 5092 blood culture isolates were collected by 29 laboratories distributed across the UK and Ireland. A single central laboratory re-identified the isolates and measured MICs by the BSAC agar dilution method. RESULTS: Oxacillin resistance was found in 42% of Staphylococcus aureus and 76% of coagulase-negative staphylococci. Streptococci were generally susceptible to beta-lactams, but tetracycline resistance was common (except in Streptococcus pneumoniae) and particularly common among group B isolates (82% resistant). Nine percent of S. pneumoniae had reduced susceptibility to penicillin (MICs 0.12-1 mg/L), but none required >/=2 mg/L for inhibition. High-level gentamicin resistance was seen in 43% of Enterococcus faecalis, often in combination with raised ciprofloxacin MICs (>/=32 mg/L), but these isolates remained susceptible to ampicillin and imipenem. Only linezolid and tigecycline showed in vitro potency against a large proportion of Enterococcus faecium. Vancomycin resistance was restricted to enterococci (20% of E. faecium, 3% of E. faecalis) and a single isolate of coagulase-negative staphylococci (0.2%, MIC of 8 mg/L). Escherichia coli isolates were commonly resistant to amoxicillin (56%) and tetracycline (88%) but remained susceptible to ceftazidime, piperacillin/tazobactam and imipenem. Extended-spectrum beta-lactamases were detected in 2% of E. coli (none in 2001, 3.2% in 2002), 5% of Klebsiella spp. and 8% of Enterobacter spp. Resistance rates of Pseudomonas aeruginosa to ciprofloxacin, ceftazidime, gentamicin, imipenem and piperacillin/tazobactam were between 4% and 7%. Among the newly licensed and developmental agents, there was no resistance to linezolid in Gram-positive organisms. Ertapenem had a wide spectrum, covering Enterobacteriaceae, streptococci and oxacillin-susceptible staphylococci. MICs of tigecycline were low for Gram-positive species and Enterobacteriaceae except Proteeae and Enterobacter spp. CONCLUSION: Antimicrobial resistance among major bloodstream pathogens to those antimicrobials often selected for empirical therapy was relatively uncommon in 2001-2002, usually <10%. An important exception was oxacillin resistance in S. aureus. PMID- 15128725 TI - Citrobacter koseri and Citrobacter amalonaticus isolates carry highly divergent beta-lactamase genes despite having high levels of biochemical similarity and 16S rRNA sequence homology. AB - OBJECTIVES: Isolates previously identified as Citrobacter diversus are now known as Citrobacter koseri. We measured sequence variation at the beta-lactamase structural gene among a group of clinical isolates originally identified as C. diversus by API 20E profiling. METHODS: beta-Lactamase and 16S rRNA genes were amplified by PCR and sequenced by standard methods. beta-Lactamase induction was attempted in liquid-grown cultures using cefoxitin. Nitrocefin hydrolysis assays were performed using a spectrophotometer. RESULTS: Analysis of 16S rRNA gene sequences showed that Citrobacter spp. isolates with an inducible beta-lactamase gene, cdiA, closely related to 'C. koseri ' NF85 and ULA27 are actually Citrobacter amalonaticus. C. koseri isolates, whose identities were confirmed by 16S rRNA sequencing, produce a class A beta-lactamase, Cko, constitutively at low levels. The cko and cdiA beta-lactamase genes share <45% identity. CONCLUSIONS: We have confirmed that cko is a beta-lactamase gene carried by C. koseri, and that isolates previously identified as 'C. koseri ', but carrying the cdiA beta lactamase gene are C. amalonaticus. Thus, beta-lactamase-gene-specific PCR may provide a valuable tool to differentiate these biochemically homogeneous Citrobacter species. PMID- 15128724 TI - Relationship between myalgias/arthralgias occurring in patients receiving quinupristin/dalfopristin and biliary dysfunction. AB - OBJECTIVES: To determine whether myalgias/arthralgias occurring in cancer patients who receive quinupristin/dalfopristin are associated with biliary tract dysfunction. METHODS: We studied 56 patients with vancomycin-resistant enterococcal infections who were treated with quinupristin/dalfopristin 7.5 mg/kg every 8 h for a mean duration of 12 days (range 2-52 days). Liver function tests, including a test for alkaline phosphatase, were performed before, during and after the end of therapy. All patients were followed for 1 month after completion of therapy. RESULTS: Thirty-eight (68%) of the 56 patients responded. Myalgias/arthralgias were the leading adverse events occurring in 20 (36%) of the patients. Patients with myalgias/arthralgias had significantly higher levels of alkaline phosphatase (mean 318.7 IU/L) during the mid-term therapy cycle compared with patients without any joint or muscular pain (mean 216.3 IU/L, P = 0.05). In addition, 3/18 (16.6%) patients with myalgias/arthralgias had more than five-fold the normal levels of alkaline phosphatase, which did not occur in any of the other patients who did not develop myalgias/arthralgias (P = 0.04). All myalgias/arthralgias resolved after the discontinuation of quinupristin/dalfopristin. By univariate analysis, other factors associated with myalgias/arthralgias were relapse of haematological malignancy (P = 0.01), receiving tacrolimus within 1 month prior to treatment (P = 0.04) and receiving methotrexate during antimicrobial therapy (P = 0.05). CONCLUSIONS: Myalgias/arthralgias occur frequently in cancer patients receiving quinupristin/dalfopristin and may be associated with biliary tract dysfunction, as measured by alkaline phosphatase or other factors that could lead to intra hepatic cholestasis, such as relapse of haematological malignancy or treatment with tacrolimus or methotrexate. PMID- 15128726 TI - Improving appropriateness of antibiotic therapy: randomized trial of an intervention to foster reassessment of prescription after 3 days. AB - OBJECTIVES: Reassessment of ongoing antibiotic therapy is an important step towards appropriate use of antibiotics. This study was conducted to evaluate the impact of a short questionnaire designed to encourage reassessment of intravenous antibiotic therapy after 3 days. PATIENTS AND METHODS: Patients hospitalized on the surgical and medical wards of a university hospital and treated with an intravenous antibiotic for 3-4 days were randomly allocated to either an intervention or control group. The intervention consisted of mailing to the physician in charge of the patient a three-item questionnaire referring to possible adaptation of the antibiotic therapy. The primary outcome was the time elapsed from randomization until a first modification of the initial intravenous antibiotic therapy. It was compared within both groups using Cox proportional hazard modelling. RESULTS: One hundred and twenty-six eligible patients were randomized in the intervention group and 125 in the control group. Time to modification of intravenous antibiotic therapy was 14% shorter in the intervention group (adjusted hazard ratio for modification 1.28, 95% CI 0.99 1.67, P = 0.06). It was significantly shorter in the intervention group compared with a similar group of 151 patients observed during a 2 month period preceding the study (adjusted hazard ratio 1.17, 95% CI 1.03-1.32, P = 0.02). CONCLUSION: The results suggest that a short questionnaire, easily adaptable to automatization, has the potential to foster reassessment of antibiotic therapy. PMID- 15128727 TI - Adjunctive efficacy of granulocyte colony-stimulating factor on treatment of Pseudomonas aeruginosa pneumonia in neutropenic and non-neutropenic hosts. AB - OBJECTIVES: Granulocyte colony-stimulating factor (G-CSF) stimulates proliferation of neutrophils and enhances their phagocytic and microcidal activity. Increasing resistance to existing antibacterials and the dearth of new alternatives have complicated the treatment of Gram-negative infections. The aim of this study was to evaluate the efficacy of G-CSF in the treatment of Pseudomonas aeruginosa pneumonia when administered in combination with ceftazidime in both neutropenic and non-neutropenic hosts. METHODS: A group of mice were rendered neutropenic with cyclophosphamide. Pneumonia was induced by intratracheal instillation of approximately 5 x 10(7) cfu/mL and approximately 5 x 10(9) cfu/mL (LD(100)) of the organism to neutropenic and non-neutropenic mice, respectively. Two hours after inoculation, the mice received normal saline and 5% dextrose, G-CSF (300 micro g/kg per day x 3 days), ceftazidime (2000 mg/kg x 2 doses) or a combination of G-CSF and ceftazidime. Survival was monitored at different time points for 5 days. RESULTS: Treatment with G-CSF showed a dose dependent increase in survival from 50 to 300 micro g/kg. In neutropenic mice, survival was markedly better in the G-CSF + ceftazidime group compared with controls (P = 0.0001), G-CSF (P = 0.0002) or ceftazidime (P = 0.0172). In non neutropenic mice, survival in the G-CSF + ceftazidime group (20%) was significantly higher than in the control and G-CSF groups (P = 0.0001) but not significantly higher than ceftazidime alone (9%) (P > 0.05). CONCLUSIONS: G-CSF administered in combination with antibiotic after onset of severe P. aeruginosa pneumonia may improve therapeutic outcome and this suggests a new treatment option in the management of pneumonia especially in neutropenic patients. PMID- 15128728 TI - CC-chemokine receptor 5 (CCR5) in hepatitis C--at the crossroads of the antiviral immune response? AB - An effective immune response to hepatitis C virus (HCV) infection requires efficient recruitment and activation of inflammatory cells to the liver, the site of infection. Chemokines are critically involved in this process, since they exert both chemotactic and immunoregulatory actions. In particular, the interaction between chemokines CCL3 (MIP-1alpha), CCL4 (MIP-1beta) and CCL5 (RANTES) and their receptor, CC-chemokine receptor 5 (CCR5), may be critical in regulating T cell functions by mediating recruitment, polarization, activation and differentiation of antiviral type 1 cytokine secreting T helper and cytotoxic T cells. A 32 bp deletion in the encoding region of CCR5 leads to complete loss of the functional CCR5 receptor in subjects homozygous for this mutation and decreased expression in heterozygous patients. This fact provides the unique opportunity to study the role of the CCR5 receptor in chronic hepatitis C infection by comparing immune responses between HCV infected CCR5-Delta32 carriers and CCR5 wild-type patients. This article will summarize and discuss the available data with respect to possibly altered disease susceptibility, clinical course and treatment outcomes associated with the CCR5-Delta32 mutation in hepatitis C. PMID- 15128729 TI - Surveillance of susceptibility patterns in 1297 European and US anaerobic and capnophilic isolates to co-amoxiclav and five other antimicrobial agents. AB - In vitro susceptibility data were collected for co-amoxiclav and other antimicrobial agents against 1297 recent anaerobe isolates collected in Europe and the USA. The co-amoxiclav (amoxicillin/clavulanic acid) MIC(50/90)s (amoxicillin/clavulanic acid concentration in a ratio of 2:1, expressed in terms of amoxicillin concentration in mg/L) were 0.5/4 for Bacteroides fragilis, 96% susceptible) except E. corrodens (MIC(50/90) of >32/>64 mg/L), which is a capnophilic organism. Imipenem was also highly active against all species (>98% susceptible). Levofloxacin and clindamycin were the least potent agents tested, particularly against Bacteroides, Prevotella and Peptostreptococcus (levofloxacin susceptibility rates: Bacteroides 72.7%, Prevotella 71.5%, F. magna 72.4%; clindamycin susceptibility rates: Bacteroides 79.5%, Prevotella 92.1%, F. magna 84.7%). PMID- 15128730 TI - Comment on: antibiotic prophylaxis, body piercing and infective endocarditis. PMID- 15128731 TI - Subcutaneous nanoparticle-based antitubercular chemotherapy in an experimental model. AB - Poly (DL-lactide-co-glycolide) (PLG) nanoparticles encapsulating three front-line antitubercular drugs, i.e. rifampicin, isoniazid and pyrazinamide, were prepared by the multiple emulsion technique and administered subcutaneously to mice for pharmacokinetic/chemotherapeutic study. A single subcutaneous dose of drug-loaded PLG nanoparticles resulted in sustained therapeutic drug levels in the plasma for 32 days and in the lungs/spleen for 36 days. The mean residence time and absolute bioavailability were increased several-fold as compared with unencapsulated drugs. Further, drug-loaded PLG nanoparticles resulted in undetectable bacterial counts in the lungs and spleen of Mycobacterium tuberculosis-infected mice, thereby demonstrating a better chemotherapeutic efficacy, as compared with daily free drug treatment. Hence, injectable PLG nanoparticles hold promise for increasing drug bioavailability and reducing dosing frequency for better management of tuberculosis. PMID- 15128732 TI - The heme synthesis defect of mutants impaired in mitochondrial iron-sulfur protein biogenesis is caused by reversible inhibition of ferrochelatase. AB - Mitochondria are responsible for the synthesis of both iron-sulfur clusters and heme, but the potential connection between the two major iron-consuming pathways is unknown. Here, we have shown that mutants in the yeast mitochondrial iron sulfur cluster (ISC) assembly machinery displayed reduced cytochrome levels and diminished activity of the heme-containing cytochrome c oxidase, in addition to iron-sulfur protein defects. In contrast, mutants in components of the mitochondrial ISC export machinery, which are specifically required for maturation of cytosolic iron-sulfur proteins, were not decreased in heme synthesis or cytochrome levels. Heme synthesis does not involve the function of mitochondrial ISC components, because immunological depletion of various ISC proteins from mitochondrial extracts did not affect the formation and amounts of heme. The heme synthesis defects of ISC mutants were found in vivo in isolated mitochondria and in mitochondrial detergent extracts and were confined to an inhibition of ferrochelatase, the enzyme catalyzing the insertion of iron into protoporphyrin IX. In support of these findings, immunopurification of ferrochelatase from ISC mutants restored its activity to wild-type levels. We conclude that the reversible inhibition of ferrochelatase is the molecular reason for the heme deficiency in ISC assembly mutants. This inhibitory mechanism may be used for regulation of iron distribution between the two iron-consuming processes. PMID- 15128733 TI - Repression of endogenous Smad7 by Ski. AB - The Ski protein has been proposed to serve as a corepressor for Smad4 to maintain a transforming growth factor-beta (TGF-beta)-responsive promoter at a repressed, basal level. However, there have been no reports so far that it indeed acts on a natural promoter. We have previously cloned the human Smad7 promoter and shown that it contains the 8-base pair palindromic Smad-binding element (SBE) necessary for TGF-beta induction. In this report, we have characterized the negative regulation of Smad7 promoter basal activity by Ski. We show that Ski inhibits the Smad7 promoter basal activity in a SBE-dependent manner. Mutation of the SBE abrogates the inhibitory effect of Ski on the Smad7 promoter. Moreover, mutation of the SBE increases the Smad7 promoter basal activity. Using the chromatin immunoprecipitation assay, we further show that Ski together with Smad4 binds to the endogenous Smad7 promoter. Finally, we show that RNAi knockdown of Ski increases Smad7 reporter gene activity in transient transfection assays as well as elevating the endogenous level of Smad7 mRNA. Taken together, our results provide the first evidence that Ski is indeed a corepressor for Smad4, which can inhibit a natural TGF-beta responsive gene at the basal state. PMID- 15128734 TI - Tryptophan 621 and serine 667 residues of Daxx regulate its nuclear export during glucose deprivation. AB - The cellular target responsible for the nuclear export of Daxx has been identified as chromosomal region maintenance 1 (CRM1), which is a carrier protein for nuclear export and a receptor for the nuclear export signal (NES) of Daxx. Binding of Daxx to CRM1 was increased early during glucose deprivation and then gradually decreased. This interaction was inhibited by leptomycin B, a specific inhibitor of CRM1-dependent nuclear export. Substitution of the serine 667 amino acid residue of Daxx with alanine reduced the interaction with CRM1 during glucose deprivation, suggesting that the phosphorylation of Ser-667 is required for its binding to CRM1 and for its subsequent nuclear export. Data from coupled transcription-translation studies reveal that the NES (amino acids 565-575) of Daxx is a binding site for CRM1. Interestingly, constitutive export of Daxx has occurred by replacement of the tryptophan 621 Daxx residue with alanine. These results suggest that this tryptophan residue plays a key role in masking the NES of Daxx from its receptor, CRM1, in the resting state, whereas phosphorylation of serine 667 would release the NES, which could then be recognized by the CRM1. PMID- 15128735 TI - Extracellular signal-regulated kinase (ERK)-dependent gene expression contributes to L1 cell adhesion molecule-dependent motility and invasion. AB - The cell adhesion molecule L1 has been implicated in a variety of motile processes, including neurite extension, cerebellar cell migration, extravasation, and metastasis. Homophilic or heterophilic L1 binding and concomitant signaling have been shown to promote cell motility in the short term. In this report, L1 is also shown to induce and maintain a motile and invasive phenotype by promoting gene transcription. In the presence of serum or platelet-derived growth factor, L1 promotes heightened and sustained activation of the extracellular signal regulated kinase pathway. Activation of this pathway then induces the expression of motility- and invasion-associated gene products, including the beta(3) integrin subunit, small GTPases, and the cysteine proteases cathepsin-L and -B. Induction of integrin alpha(v)beta(3) and rac-1 is shown to contribute directly to L1-dependent haptotaxis, whereas induction of cathepsins-L and -B promotes matrix invasion. This study provides a novel translational mechanism to account for the association between L1 expression and motile processes involved in metastasis and development. PMID- 15128736 TI - Evidence for an unfolding/threading mechanism for protein disaggregation by Saccharomyces cerevisiae Hsp104. AB - Saccharomyces cerevisiae Hsp104, a hexameric member of the Hsp100/Clp subfamily of AAA+ ATPases with two nucleotide binding domains (NBD1 and 2), refolds aggregated proteins in conjunction with Hsp70 molecular chaperones. Hsp104 may act as a "molecular crowbar" to pry aggregates apart and/or may extract proteins from aggregates by unfolding and threading them through the axial channel of the Hsp104 hexamer. Targeting Tyr-662, located in a Gly-Tyr-Val-Gly motif that forms part of the axial channel loop in NBD2, we created conservative (Phe and Trp) and non-conservative (Ala and Lys) amino acid substitutions. Each of these Hsp104 derivatives was comparable to the wild type protein in their ability to hydrolyze ATP, assemble into hexamers, and associate with heat-shock-induced aggregates in living cells. However, only those with conservative substitutions complemented the thermotolerance defect of a Deltahsp104 yeast strain and promoted refolding of aggregated protein in vitro. Monitoring fluorescence from Trp-662 showed that titration of fully assembled molecules with either ATP or ADP progressively quenches fluorescence, suggesting that nucleotide binding determines the position of the loop within the axial channel. A Glu to Lys substitution at residue 645 in the NBD2 axial channel strongly alters the nucleotide-induced change in fluorescence of Trp-662 and specifically impairs in protein refolding. These data establish that the structural integrity of the axial channel through NBD2 is required for Hsp104 function and support the proposal that Hsp104 and ClpB use analogous unfolding/threading mechanisms to promote disaggregation and refolding that other Hsp100s use to promote protein degradation. PMID- 15128737 TI - Comprehensive chimeric analysis of amino acid residues critical for high affinity glucose transport by Hxt2 of Saccharomyces cerevisiae. AB - Chimeras of Hxt2 and Hxt1, high affinity and low affinity glucose transporters, respectively, of Saccharomyces cerevisiae, were previously constructed by random replacement of each of the 12 transmembrane segments (TMs) of Hxt2 with the corresponding region of Hxt1. Characterization of these chimeras revealed that at least TMs 1, 5, 7, and 8 of Hxt2 are required for high affinity transport activity. To determine which amino acid residues in these TMs are important for high affinity glucose transport, we systematically shuffled all of the 20 residues in these regions that differ between Hxt2 and Hxt1. Analysis of 60 independent mutant strains identified as expressing high affinity and high capacity glucose transport activity by selection on glucose-limited agar plates revealed that Leu-201 in TM5 of Hxt2 is most important for such activity and that either Cys-195 or Phe-198 is also required for maximal activity. PMID- 15128738 TI - Atomic resolution structures and solution behavior of enzyme-substrate complexes of Enterobacter cloacae PB2 pentaerythritol tetranitrate reductase. Multiple conformational states and implications for the mechanism of nitroaromatic explosive degradation. AB - The structure of pentaerythritol tetranitrate (PETN) reductase in complex with the nitroaromatic substrate picric acid determined previously at 1.55 A resolution indicated additional electron density between the indole ring of residue Trp-102 and the nitro group at C-6 of picrate. The data suggested the presence of an unusual bond between substrate and the tryptophan side chain. Herein, we have extended the resolution of the PETN reductase-picric acid complex to 0.9 A. This high-resolution analysis indicates that the active site is partially occupied with picric acid and that the anomalous density seen in the original study is attributed to the population of multiple conformational states of Trp-102 and not a formal covalent bond between the indole ring of Trp-102 and picric acid. The significance of any interaction between Trp-102 and nitroaromatic substrates was probed further in solution and crystal complexes with wild-type and mutant (W102Y and W102F) enzymes. Unlike with wild-type enzyme, in the crystalline form picric acid was bound at full occupancy in the mutant enzymes, and there was no evidence for multiple conformations of active site residues. Solution studies indicate tighter binding of picric acid in the active sites of the W102Y and W102F enzymes. Mutation of Trp-102 does not impair significantly enzyme reduction by NADPH, but the kinetics of decay of the hydride Meisenheimer complex are accelerated in the mutant enzymes. The data reveal that decay of the hydride-Meisenheimer complex is enzyme catalyzed and that the final distribution of reaction products for the mutant enzymes is substantially different from wild-type enzyme. Implications for the mechanism of high explosive degradation by PETN reductase are discussed. PMID- 15128739 TI - Recycling and resensitization of the neurokinin 1 receptor. Influence of agonist concentration and Rab GTPases. AB - Substance P (SP) induces endocytosis and recycling of the neurokinin 1 receptor (NK1R) in endothelial cells and spinal neurons at sites of inflammation and pain, and it is thus important to understand the mechanism and function of receptor trafficking. We investigated how the SP concentration affects NK1R trafficking and determined the role of Rab GTPases in trafficking. NK1R trafficking was markedly influenced by the SP concentration. High SP (10 nM) induced translocation of the NK1R and beta-arrestin 1 to perinuclear sorting endosomes containing Rab5a, where NK1R remained for >60 min. Low SP (1 nM) induced translocation of the NK1R to early endosomes located immediately beneath the plasma membrane that also contained Rab5a and beta-arrestin 1, followed by rapid recycling of the NK1R. Overexpression of Rab5a promoted NK1R translocation to perinuclear sorting endosomes, whereas the GTP binding-deficient mutant Rab5aS34N caused retention of the NK1R in superficial early endosomes. NK1R translocated from superficial early endosomes to recycling endosomes containing Rab4a and Rab11a, and Rab11aS25N inhibited NK1R recycling. Rapid NK1R recycling coincided with resensitization of SP-induced Ca2+ mobilization and with the return of surface SP binding sites. Resensitization was minimally affected by inhibition of vacuolar H(+)-ATPase and phosphatases but was markedly suppressed by disruption of Rab4a and Rab11a. Thus, whereas beta-arrestins mediate NK1R endocytosis, Rab5a regulates translocation between early and sorting endosomes, and Rab4a and Rab11a regulate trafficking through recycling endosomes. We have thus identified a new function of Rab5a as a control protein for directing concentration-dependent trafficking of the NK1R into different intracellular compartments and obtained evidence that Rab4a and Rab11a contribute to G-protein-coupled receptor recycling from early endosomes. PMID- 15128740 TI - Kinetic and mechanistic studies of a cell cycle protein phosphatase Cdc14. AB - The Cdc14 family of protein phosphatases is conserved within eukaryotes and antagonizes the action of cyclin-dependent kinases, thereby promoting mitotic exit and cytokinesis. We performed a detailed kinetic and mechanistic study of the Cdc14 phosphatases with both small molecule aryl phosphates and a physiological protein substrate hCdh1. We found that Cdc14 displays a strong preference for two-ringed aryl phosphates over smaller one-ringed or larger, multi-ringed substrates, a finding that may have important implications for inhibitor design. Results from both leaving group and pH dependence of the Cdc14 catalyzed reaction are consistent with a general acid-independent mechanism for substrates with leaving group pKa < 7 and a general acid-dependent mechanism for substrates with leaving group pKa > 7. The use of both low and high leaving group pKa substrates, in combination with steady-state and pre-steady-state kinetic techniques enabled the isolation and analysis of both the phosphoenzyme (E-P) formation and hydrolysis step. We established the requirement of general acid catalysis for E-P formation in reactions with high leaving group pKa substrates, and the presence of general base catalysis in E-P hydrolysis. Mutational study of invariant acidic residues in Cdc14 identified Asp253 as the general acid during E P formation and the general base in E-P hydrolysis. We also identified several residues including Asp50, Asp129, Glu168, Glu171, and Asp177 in the Cdc14 active site cleft that are required for efficient dephosphorylation of hCdh1. PMID- 15128741 TI - Tumor necrosis factor-alpha and CD80 modulate CD28 expression through a similar mechanism of T-cell receptor-independent inhibition of transcription. AB - Replicative senescence of human T cells is characterized by the loss of CD28 expression, exemplified by the clonal expansion of CD28(null) T cells during repeated stimulation in vitro as well as in chronic inflammatory and infectious diseases and in the normal course of aging. Because CD28 is the major costimulatory receptor for the induction of T cell-mediated immunity, the mechanism(s) underlying CD28 loss is of paramount interest. Current models of replicative senescence involve protracted procedures to generate CD28(null) cells from CD28(+) precursors; hence, a T-cell line model was used to examine the dynamics of CD28 expression. Here, we show the versatility of the JT and Jtag cell lines in tracking CD28(null) <--> CD28(hi) phenotypic transitions. JT and Jtag cells were CD28(null) and CD28(lo), respectively, but expressed high levels of CD28 when exposed to phorbol 12-myristate 13-acetate. This was a result of the reconstitution of the CD28 gene transcriptional initiator (INR). Tumor necrosis factor-alpha reduced CD28 expression because of the inhibition of INR-driven transcription. Ligation of CD28 by an antibody or by CD80 also down-regulated CD28 transcription through the same mechanism, providing evidence that CD28 can generate a T cell receptor-independent signal with a unique biological outcome. Collectively, these data unequivocally demonstrate the critical role of the INR in the regulation of CD28 expression. T cell lines with transient expression of CD28 are invaluable in the dissection of the biochemical processes involved in the transactivation of the CD28 INR, the silencing of which is a key event in the ontogenesis of senescent T cells. PMID- 15128742 TI - Functional and structural diversity in the Als protein family of Candida albicans. AB - The human fungal pathogen Candida albicans colonizes and invades a wide range of host tissues. Adherence to host constituents plays an important role in this process. Two members of the C. albicans Als protein family (Als1p and Als5p) have been found to mediate adherence; however, the functions of other members of this family are unknown. In this study, members of the ALS gene family were cloned and expressed in Saccharomyces cerevisiae to characterize their individual functions. Distinct Als proteins conferred distinct adherence profiles to diverse host substrates. Using chimeric Als5p-Als6p constructs, the regions mediating substrate-specific adherence were localized to the N-terminal domains in Als proteins. Interestingly, a subset of Als proteins also mediated endothelial cell invasion, a previously unknown function of this family. Consistent with these results, homology modeling revealed that Als members contain anti-parallel beta sheet motifs interposed by extended regions, homologous to adhesins or invasins of the immunoglobulin superfamily. This finding was confirmed using circular dichroism and Fourier transform infrared spectrometric analysis of the N-terminal domain of Als1p. Specific regions of amino acid hypervariability were found among the N-terminal domains of Als proteins, and energy-based models predicted similarities and differences in the N-terminal domains that probably govern the diverse function of Als family members. Collectively, these results indicate that the structural and functional diversity within the Als family provides C. albicans with an array of cell wall proteins capable of recognizing and interacting with a wide range of host constituents during infection. PMID- 15128743 TI - Structural and functional characterization of a novel phosphodiesterase from Methanococcus jannaschii. AB - Methanococcus jannaschii MJ0936 is a hypothetical protein of unknown function with over 50 homologs found in many bacteria and Archaea. To help define the molecular (biochemical and biophysical) function of MJ0936, we determined its crystal structure at 2.4-A resolution and performed a series of biochemical screens for catalytic activity. The overall fold of this single domain protein consists of a four-layered structure formed by two beta-sheets flanked by alpha helices on both sides. The crystal structure suggested its biochemical function to be a nuclease, phosphatase, or nucleotidase, with a requirement for some metal ions. Crystallization in the presence of Ni(2+) or Mn(2+) produced a protein containing a binuclear metal center in the putative active site formed by a cluster of conserved residues. Analysis of MJ0936 against a panel of general enzymatic assays revealed catalytic activity toward bis-p-nitrophenyl phosphate, an indicator substrate for phosphodiesterases and nucleases. Significant activity was also found with two other phosphodiesterase substrates, thymidine 5' monophosphate p-nitrophenyl ester and p-nitrophenylphosphorylcholine, but no activity was found for cAMP or cGMP. Phosphodiesterase activity of MJ0936 had an absolute requirement for divalent metal ions with Ni(2+) and Mn(2+) being most effective. Thus, our structural and enzymatic studies have identified the biochemical function of MJ0936 as that of a novel phosphodiesterase. PMID- 15128744 TI - The intrinsic threshold of the fibrinolytic system is modulated by basic carboxypeptidases, but the magnitude of the antifibrinolytic effect of activated thrombin-activable fibrinolysis inhibitor is masked by its instability. AB - Activated thrombin-activable fibrinolysis inhibitor (TAFIa) is intrinsically unstable, a property that complicates the study of its role in regulating fibrinolysis. To investigate the effect of basic carboxypeptidases on fibrinolysis under conditions of constant carboxypeptidase activity, we employed pancreatic carboxypeptidase B (CPB), a homologous, stable basic carboxypeptidase, as a surrogate for TAFIa. Clots formed from TAFI-depleted plasma or from purified components were supplemented with tissue-type plasminogen activator and either CPB or TAFIa. The clot lysis data indicate that the down-regulation of fibrinolysis mediated by basic carboxypeptidases involves a threshold mechanism. At carboxypeptidase concentrations above the threshold, plasminogen activation is maintained in a fully down-regulated state; experiments in plasma showed that fibrinolysis is essentially halted by saturating concentrations of TAFIa and that fibrinolysis can be prolonged more than 45-fold by a stable carboxypeptidase. The threshold carboxypeptidase concentration was dependent on tissue-type plasminogen activator and antiplasmin concentrations, indicating that the threshold is determined by the steady-state plasmin concentration. Although obvious with CPB, the threshold was masked by the intrinsic instability of TAFIa and became apparent only when the effect of TAFIa was investigated over the picomolar concentration range. Because of the threshold effect and the instability of TAFIa, exponential increases in TAFIa concentration generate linear increases in lysis time. A model relating lysis time to TAFIa concentration, TAFIa half-life, and the threshold concentration of TAFIa is provided. The threshold effect has potentially important implications regarding the role of TAFIa and the regulation of clot lysis in vivo. PMID- 15128745 TI - Hyperglycemia promotes oxidative stress through inhibition of thioredoxin function by thioredoxin-interacting protein. AB - Increased intracellular reactive oxygen species (ROS) contribute to vascular disease and pro-atherosclerotic effects of diabetes mellitus may be mediated by oxidative stress. Several ROS-scavenging systems tightly control cellular redox balance; however, their role in hyperglycemia-induced oxidative stress is unclear. A ubiquitous antioxidative mechanism for regulating cellular redox balance is thioredoxin, a highly conserved thiol reductase that interacts with an endogenous inhibitor, thioredoxin-interacting protein (Txnip). Here we show that hyperglycemia inhibits thioredoxin ROS-scavenging function through p38 MAPK mediated induction of Txnip. Overexpression of Txnip increased oxidative stress, while Txnip gene silencing restored thioredoxin activity in hyperglycemia. Diabetic animals exhibited increased vascular expression of Txnip and reduced thioredoxin activity, which normalized with insulin treatment. These results provide evidence for the impairment of a major ROS-scavenging system in hyperglycemia. These studies implicate reduced thioredoxin activity through interaction with Txnip as an important mechanism for vascular oxidative stress in diabetes mellitus. PMID- 15128746 TI - Impact of the mitogen-activated protein kinase pathway on parathyroid hormone related protein actions in osteoblasts. AB - Parathyroid hormone-related protein (PTHrP) regulates proliferation and differentiation of osteoblastic cells via binding to the parathyroid hormone receptor (PTH-1R). The cAMP-dependent protein kinase A pathway governs the majority of these effects, but recent evidence also implicates the MAPK pathway. MC3T3-E1 subclone 4 cells (MC4) were treated with the MAPK inhibitor U0126 and PTHrP. In differentiated MC4 cells, osteocalcin and bone sialoprotein gene expression were both down-regulated by PTHrP and also by inhibition of the MAPK pathway. PTHrP-mediated down-regulation of PTH-1R mRNA and up-regulation of c-fos mRNA were MAPK-independent, whereas PTHrP stimulation of fra-2 and interleukin-6 (IL-6) mRNA was MAPK-dependent. Luciferase promoter assays revealed that regulation of IL-6 involved the cAMP-dependent protein kinase A and MAPK pathways with a potential minor role of the protein kinase C pathway, and a promoter region containing an activator protein-1 site was necessary for PTHrP-induced IL 6 gene transcription. An alternative pathway, through cAMP/Epac/Rap1/MAPK, mediated ERK phosphorylation but was not sufficient for IL-6 promoter activation. Phosphorylation of the transcription factor CREB was also necessary but not sufficient for PTHrP-mediated IL-6 promoter activity. Most interesting, a bidirectional effect was found with PTHrP increasing phosphorylated ERK in undifferentiated MC4 cells but decreasing phosphorylated ERK in differentiated cells. These data indicate that inactivation of the MAPK pathway shows differential regulation of PTHrP-stimulated activator protein-1 members, blocks PTHrP-stimulated IL-6, and synergistically down-regulates certain osteoblastic markers associated with differentiation. These novel findings indicate that the MAPK pathway plays a selective but important role in the actions of PTHrP. PMID- 15128747 TI - Multiple molecular determinants in the carboxyl terminus regulate dopamine transporter export from endoplasmic reticulum. AB - The plasma membrane dopamine transporter (DAT) has an essential role in terminating dopaminergic neurotransmission by reuptake of dopamine into the presynaptic neurons. Therefore, the amount of DAT at the cell surface is a critical determinant of DAT function. In this study, we examined the role of the carboxyl terminus of DAT in trafficking of the transporter through the biosynthetic pathway to the plasma membrane. Live cell fluorescence microscopy and cell surface biotinylation were used to study the effects of systematic deletions and alanine substitutions in the carboxyl terminus on DAT localization. It was found that alanine substitutions of Lys-590 and Asp-600 significantly delayed the delivery of DAT to the plasma membrane because of retention of DAT in the endoplasmic reticulum (ER). Most surprising, mutation of Gly-585 to alanine completely blocked the exit of DAT from the ER and surface expression of the transporter. The effect of these three mutations on ER export of DAT was demonstrated in porcine aortic endothelial cells and the immortalized neuronal cell line 1RB3AN27. In primary cultures of rat embryonic midbrain neurons, DAT G585A, K590A, and D600A mutants were restricted to the cell soma and did not traffic to the dendrites or axonal processes. These data are consistent with the model whereby the local conformation and/or intramolecular interactions of the sequences of the DAT carboxyl terminus proximal to the last transmembrane domain are essential for the ER export of the transporter. PMID- 15128748 TI - Refolding processes of cytochrome P450cam from ferric and ferrous acid forms to the native conformation. Formations of folding intermediates with non-native heme coordination state. AB - Changes in heme coordination state and protein conformation of cytochrome P450(cam) (P450(cam)), a b-type heme protein, were investigated by employing pH jump experiments coupled with time-resolved optical absorption, fluorescence, circular dichroism, and resonance Raman techniques. We found a partially unfolded form (acid form) of ferric P450(cam) at pH 2.5, in which a Cys(-)-heme coordination bond in the native conformation was ruptured. When the pH was raised to pH 7.5, the acid form refolded to the native conformation through a distinctive intermediate. Formations of similar acid and intermediate forms were also observed for ferrous P450(cam). Both the ferric and ferrous forms of the intermediate were found to have an unidentified axial ligand of the heme at the 6th coordination sphere, which is vacant in the high spin ferric and ferrous forms at the native conformation. For the ferrous form, it was also indicated that the 5th axial ligand is different from the native cysteinate. The folding intermediates identified in this study demonstrate occurrences of non-native coordination state of heme during the refolding processes of the large b-type heme protein, being akin to the well known folding intermediates of cytochromes c, in which c-type heme is covalently attached to a smaller protein. PMID- 15128749 TI - Chemokines generally exhibit scavenger receptor activity through their receptor binding domain. AB - Chemokines are a family of cytokines that induce directed migration of various types of leukocytes through specific interactions with a group of seven transmembrane receptors. Scavenger receptors are a heterogenous family of transmembrane molecules that commonly bind and uptake oxidized low density lipoprotein and bacteria. Here, we show that not only CXC chemokine 16 (CXCL16)/SR-PSOX, a transmembrane chemokine with scavenger receptor activity, but also 12 out of 15 chemokines examined efficiently bound scavenger receptor ligands in competition with cells expressing their specific chemokine receptors. Furthermore both the chemotactic and scavenger receptor activities of SR PSOX/CXCL16 were similarly impaired in a series of mutants altered in the chemokine domain, indicating that SR-PSOX/CXCL16 binds scavenger receptor ligands as well as CXCR6 using highly overlapping binding motifs. Taken together, chemokines generally have scavenger receptor-like activity through their receptor binding domain, suggesting a close evolutionary relationship between chemokines and scavenger receptors. PMID- 15128750 TI - Neural sensitivity to periodicity in the inferior colliculus: evidence for the role of cochlear distortions. AB - Responses of low characteristic-frequency (CF) neurons in the inferior colliculus were obtained to amplitude-modulated (AM) high-frequency tones in which the modulation rate was equal to the neuron's CF. Despite all spectral components lying outside the pure tone-evoked response areas, discharge rates were modulated by the AM signals. Introducing a low-frequency tone (CF - 1 Hz) to the same ear as the AM tones produced a 1-Hz beat in the neural response. Introducing a tone (CF - 1 Hz) to the opposite ear to the AM tone also produced a beat in the neural response, with the beat at the period of the interaural phase difference between the CF - 1 Hz tone in one ear, and the AM rate in the other ear. The monaural and interaural interactions of the AM signals with introduced pure tones suggest that AM tones generate combination tones, (inter-modulation distortion) on the basilar membrane. These interact with low-frequency tones presented to the same ear to produce monaural beats on the basilar membrane, modulating the responses of inferior colliculus (IC) neurons on the 1-Hz period of the monaural beats or interacting binaurally with neural input generated in response to stimulation of the opposite ear. The auditory midbrain appears to show a robust representation of cochlear distortions generated by amplitude-modulated sounds. PMID- 15128751 TI - Contrast-dependent spatial summation in the lateral geniculate nucleus and retina of the cat. AB - Based on extracellular recordings from 69 lateral geniculate nucleus (LGN) cells in the anesthetized cat, we found spatial summation within their receptive fields to be dependent on the contrast of the stimuli presented. By fitting the summation curves to a difference of Gaussians model, we attributed this contrast dependent effect to an actual change in the size of the center mechanism. Analogous changes in spatial frequency tuning were also observed, specifically increased peaks and cut-off frequencies with contrast. These effects were seen across the populations of both X and Y cell types. In a few cases, LGN cells were recorded simultaneously with one of their retinal ganglion cell (RGC) inputs (S potentials). In every case, the RGCs exhibited similar contrast-dependent effects in the space and spatial-frequency domains. We propose that this contrast dependency in the retinal ganglion cells results directly from a reduction in the size of the center mechanism due to an increase in contrast. We also propose that these properties first arise in the retina and are transmitted passively through the LGN to visual cortex. PMID- 15128752 TI - Firing properties and connectivity of neurons in the rat lateral central nucleus of the amygdala. AB - Using whole cell recordings from acute slices of the rat amygdala, we have examined the physiological properties of and synaptic connectivity to neurons in the lateral sector of the central amygdala (CeA). Based on their response to depolarizing current injections, CeA neurons could be divided into three types. Adapting neurons fired action potentials at the start of the current injections at high frequency and then showed complete spike-frequency adaptation with only six to seven action potentials evoked with suprathreshold current injections. Late-firing neurons fired action potentials with a prolonged delay at threshold but then discharged continuously with larger current injections. Repetitive firers discharged at the start of the current injection at threshold and then discharged continuously with larger current injections. All three cells showed prolonged afterhyperpolarizations (AHPs) that followed trains of action potentials. The AHP was longer lasting with a larger slow component in adapting neurons. The AHP in all cell types contained a fast component that was inhibited by the SK channel blocker UCL1848. The slow component, not blocked by UCL1848, was blocked by isoprenaline and was significantly larger in adapting neurons. Blockade of SK channels increased the discharge frequency in late firers and regular-spiking neurons but had no effect on adapting neurons. Blockade of the slow AHP with isoprenaline had no effect on any cell type. All cells received a mixed glutamatergic and GABAergic input from a medial pathway. Electrical stimulation of the lateral (LA) and basolateral (BLA) nuclei evoked a large monosynaptic glutamatergic response followed by a disynaptic inhibitory postsynaptic potential. Activation of neurons in the LA and BLA by puffer application of glutamate evoked a small monosynaptic response in 13 of 55 CeA neurons. Local application of glutamate to the CeL evoked a GABAergic response in all cells. These results show that at least three types of neurons are present in the CeA that can be distinguished on their firing properties. The firing frequency of two of these cell types is determined by activation of SK channels. Cells receive a small input from the LA and BLA but may receive inputs that course through these nuclei en route to the CeA. PMID- 15128754 TI - Theta rhythmic stimulation of stratum lacunosum-moleculare in rat hippocampus contributes to associative LTP at a phase offset in stratum radiatum. AB - Computational modeling demonstrates that encoding and context-dependent retrieval of memories in region CA1 of the hippocampus will be most effective when the phase of strongest entorhinal input (to stratum lacunosum-moleculare) is offset from the phase of maximal induction of long-term potentiation at Schaffer collateral synapses (in s. radiatum). This would allow entorhinal input to play a role in both retrieval and encoding without engaging long-term potentiation (LTP) during retrieval. Experiments in brain slice preparations of the hippocampal formation tested the relationship between rhythmic input to s. lacunosum moleculare and the time of maximal LTP induction at Schaffer collateral synapses in s. radiatum. Analysis of the data demonstrates a statistically significant difference in the induction of LTP for different time intervals between the end of each four-pulse train in s. lacunosum-moleculare and the single pulse s. radiatum stimulation. The time of maximal LTP induction was found to be approximately 30 ms after the end of lacunosum-moleculare stimulation, consistent with the requirements of the model. PMID- 15128753 TI - In vivo mammalian brain imaging using one- and two-photon fluorescence microendoscopy. AB - One of the major limitations in the current set of techniques available to neuroscientists is a dearth of methods for imaging individual cells deep within the brains of live animals. To overcome this limitation, we developed two forms of minimally invasive fluorescence microendoscopy and tested their abilities to image cells in vivo. Both one- and two-photon fluorescence microendoscopy are based on compound gradient refractive index (GRIN) lenses that are 350-1,000 microm in diameter and provide micron-scale resolution. One-photon microendoscopy allows full-frame images to be viewed by eye or with a camera, and is well suited to fast frame-rate imaging. Two-photon microendoscopy is a laser-scanning modality that provides optical sectioning deep within tissue. Using in vivo microendoscopy we acquired video-rate movies of thalamic and CA1 hippocampal red blood cell dynamics and still-frame images of CA1 neurons and dendrites in anesthetized rats and mice. Microendoscopy will help meet the growing demand for in vivo cellular imaging created by the rapid emergence of new synthetic and genetically encoded fluorophores that can be used to label specific brain areas or cell classes. PMID- 15128755 TI - Purkinje cell spike firing in the posterolateral cerebellum: correlation with visual stimulus, oculomotor response, and error feedback. AB - Complex (CS)- and simple-spike (SS) discharge from single Purkinje cells (Pc) in the posterolateral cerebellum of two monkeys was recorded during a visually guided reach-touch task. A visual target appeared (TA) off-gaze at a random location on a screen. On initiation of arm reach, the target disappeared, then reappeared (TR) after a fixed delay. TR was either at the same location (baseline condition) or a shifted location at a fixed distance and direction from TA location (shift condition). Across trials, we observed one or two peaks of CS activity, depending on the reach condition. The first CS (T1 CS) peak was tuned to the location of TA on the screen, following TA by approximately 150 ms. The second CS (T2 CS) peak occurred only in the shift condition, was tuned to the shift location of TR, and followed TR by approximately 150 ms. The locational preferences of T1 and T2 CS peaks were the same. T1 and T2 CSs preceded saccades to TA and TR at the preferred location and occurred during reaches with either arm. T1 CSs occurred during trials in which the target appeared, and there was a saccade to target, but no subsequent arm reach followed. SS firing varied with TA/TR in the same preferred location as for the accompanying CS. We conclude that posterolateral Pc CS and SS firing changes following an off-gaze visual target appearance in a preferred location when there is a subsequent saccade to that location. PMID- 15128757 TI - A. actinomycetemcomitans cytolethal distending toxin. PMID- 15128758 TI - DNA acrobats of the Ig class switch. AB - Small resting B lymphocytes all start out producing IgM Abs. Upon encountering Ag, the cells become activated and make a switch from IgM to other Ig classes. This class switch serves to distribute a particular V region to different Ig C regions. Each C region mediates a specialized effector function, and so, through switching, an organism can guide its Abs to various sites. Creating the new H chain requires loop-out and deletion of DNA between switch regions. These DNA acrobatics require transcription of the switch regions, presumably so that necessary factors can gain access to the DNA. These requisite switching factors include activation-induced cytidine deaminase and components of general DNA repair, including base excision repair, mismatch repair, and double-strand break repair. Despite much recent progress, not all important factors have been discovered, especially those that may guide recombination to a particular subclass. PMID- 15128759 TI - Cutting edge: the natural ligand for glucocorticoid-induced TNF receptor-related protein abrogates regulatory T cell suppression. AB - CD4(+)25(+) regulatory T (Treg) cells maintain immunological self-tolerance through mechanisms that are only in part understood. Previous studies suggest that the glucocorticoid-induced TNFR-related protein (GITR), which is preferentially expressed on the surface of Treg cells, potentially provides a signal that abrogates Treg suppression. In this study, we show that a soluble form of mouse GITR ligand (sGITR-L) induces GITR-dependent NF-kappaB activation and blocks in vitro suppression mediated by both resting and preactivated polyclonal and Ag-specific Treg cells. Since sGITR-L along with rIL-2 induces proliferation of CD4(+)25(+) cells, it appears that sGITR-L can break the anergic state of Treg cells. Because sGITR-L also up-regulates IL-2 secretion by activated CD4(+)25 (-)T cells, these two sGITR-L induced signals synergize to interfere with suppressor activity by CD4(+)25(+) Treg cells. PMID- 15128760 TI - Cutting edge: extracellular signal-regulated kinases 1/2 function as integrators of TCR signal strength. AB - Altered signaling through the TCR is currently showing promise for immunotherapy. However, the molecular mechanisms are not completely understood. Therefore, we investigated whether varying the strength of TCR engagement in various human T cells would yield different second messenger responses. The kinetics and duration of extracellular signal-regulated kinase (ERK) activation, central to multiple cellular responses, are distinctly dependent on the T cell activation state (naive vs effector), strength of TCR cross-linking, and input from the phosphatidylinositol-3 kinase pathway, which is regulated by cytokines and growth factors. Moreover, the duration of ERK activation affects c-Fos expression, a component of the AP-1 transcription complex. Thus, the character of ERK activation, transient or sustained, acts as a signal integrator to quantify the strength of TCR engagement and direct the cellular response. PMID- 15128761 TI - Cutting edge: C3d functions as a molecular adjuvant in the absence of CD21/35 expression. AB - Complement component C3 covalently attaches to Ags following activation, where the C3d cleavage fragment can function as a molecular adjuvant to augment humoral immune responses. C3d is proposed to exert its adjuvant-like activities by targeting Ags to the C3d receptor (CD21/35) expressed by B cells and follicular dendritic cells. To directly assess the importance of CD21/35 in mediating the immunostimulatory effects of C3d, CD21/35-deficient (CD21/35(-/-)) mice were immunized with streptavidin (SA), SA-C3dg tetramers, recombinant HIV gp120 (gp120), or gp120 fused with linear multimers of C3d. Remarkably, SA- and gp120 specific Ab responses were significantly augmented in CD21/35(-/-) mice when these Ags were complexed with C3d in comparison to Ag alone. In fact, primary and secondary Ab responses and Ab-forming cell responses of CD21/35(-/-) mice approached those of wild-type mice immunized with SA-C3dg and gp120-C3d. Thus, C3d can function as a molecular adjuvant in the absence of CD21/35 expression. PMID- 15128762 TI - Cutting edge: TREM-like transcript-1, a platelet immunoreceptor tyrosine-based inhibition motif encoding costimulatory immunoreceptor that enhances, rather than inhibits, calcium signaling via SHP-2. AB - To date, immunoreceptor tyrosine-based inhibition motifs (ITIMs) have been shown to mediate inhibitory properties. We report a novel triggering receptor expressed on myeloid cells (TREM) family member, TREM-like transcript-1 (TLT1), which differs from the activating members because its cytoplasmic tail contains two ITIMs at Y245 and Y281. A TLT1 splice variant (TLT1sp) encodes a different cytoplasmic tail lacking ITIMs. Both isoforms are expressed in resting platelet alpha-granules, which are up-regulated to the cell surface following activation. TLT1 recruited Src homology 2 domain-containing tyrosine phosphatase (SHP)-2 to the "classical" ITIM (Y281) but not the "nonclassical" ITIM (Y245). In contrast to previously characterized ITIM receptors, TLT1 enhanced, rather than inhibited, FcepsilonRI-mediated calcium signaling in rat basophilic leukemia cells, a property dependent on the SHP-2 recruiting classical Y281 ITIM. Therefore, TLT1 represents a new costimulatory ITIM immunoreceptor and is the second ITIM-bearing receptor to be identified in platelets after platelet endothelial cell adhesion molecule-1. PMID- 15128763 TI - Shift from systemic to site-specific memory by tumor-targeted IL-2. AB - IL-2 has been approved for treatment of patients with cancer. Moreover, it has been used as a component of vaccines against cancer. In this regard, we have recently demonstrated that dendritic cell-based peptide vaccination in mice required IL-2 to mount an effective immune response against established melanoma metastases. In this study, we confirm this observation by use of tumor-targeted IL-2. However, the development of a protective systemic memory was substantially impaired by this measure, i.e., mice, which successfully rejected s.c. tumors of B16 melanoma after vaccination with dendritic cells pulsed with tyrosinase related protein 2-derived peptides plus a boost with targeted IL-2, failed to reject a rechallenge with experimental pulmonary metastases. Detailed analysis revealed a change in the distribution of the tumor-reactive T cell population: although targeted IL-2 expanded the local effector population, tyrosinase-related protein 2-reactive T cells were almost completely depleted from lymphatic tissues. PMID- 15128764 TI - Evolutionary origins of lymphocytes: ensembles of T cell and B cell transcriptional regulators in a cartilaginous fish. AB - The evolutionary origins of lymphocytes can be traced by phylogenetic comparisons of key features. Homologs of rearranging TCR and Ig (B cell receptor) genes are present in jawed vertebrates, but have not been identified in other animal groups. In contrast, most of the transcription factors that are essential for the development of mammalian T and B lymphocytes belong to multigene families that are represented by members in the majority of the metazoans, providing a potential bridge to prevertebrate ancestral roles. This work investigates the structure and regulation of homologs of specific transcription factors known to regulate mammalian T and B cell development in a representative of the earliest diverging jawed vertebrates, the clearnose skate (Raja eglanteria). Skate orthologs of mammalian GATA-3, GATA-1, EBF-1, Pax-5, Pax-6, Runx2, and Runx3 have been characterized. GATA-3, Pax-5, Runx3, EBF-1, Spi-C, and most members of the Ikaros family are shown throughout ontogeny to be 1) coregulated with TCR or Ig expression, and 2) coexpressed with each other in combinations that for the most part correspond to known mouse T and B cell patterns, supporting conservation of function. These results indicate that multiple components of the gene regulatory networks that operate in mammalian T cell and B cell development were present in the common ancestor of the mammals and the cartilaginous fish. However, certain factors relevant to the B lineage differ in their tissue-specific expression patterns from their mouse counterparts, suggesting expanded or divergent B lineage characteristics or tissue specificity in these animals. PMID- 15128765 TI - CpG motifs as proinflammatory factors render autochthonous tumors permissive for infiltration and destruction. AB - In a transgenic mouse model expressing SV40 T Ag (Tag) as a de novo tumor Ag, immune surveillance fails and islet cell carcinomas grow progressively. To develop an anticancer strategy that would be effective in eradicating solid, autochthonously growing tumors, we evaluated the effectiveness of immunostimulatory oligodeoxynucleotides (ODN) with cytosine-guanine-rich (CpG) motifs (CpG-ODN). In a classical vaccination protocol, Tag was administered with CpG-ODN as adjuvant. The antitumor vaccination, however, was only effective in a prophylactic setting, despite the successful activation of a Tag-specific CTL response in vivo. Histological examination demonstrated that even primed immune cells failed to infiltrate tumors once a malignant environment was established. To ensure that effector cells were not limiting, highly activated tumor Ag specific T cells were transferred into tumor-bearing mice. However, this treatment also failed to result in tumor infiltration and rejection. Therefore, we further tested the efficacy of CpG-ODN as a proinflammatory agent in combination with the transfer of preactivated Tag-specific CD4(+) and CD8(+) T cells. Indeed, this combination therapy proved to be highly effective, because CpG-ODN rendered insulinomas permissive for massive infiltration and destruction. The opening of tumor tissue correlated with uptake of CpG-ODN by tissue-resident macrophages and a strong up-regulation of adhesion molecules such as ICAM and VCAM on blood vessel endothelia. These data demonstrate that systemic application of proinflammatory reagents drastically enhances extravasation of effector cells into tumor tissue, an observation that is of general importance for immunotherapy of solid tumors in a clinical setting. PMID- 15128766 TI - Differential gene expression patterns by oligonucleotide microarray of basal versus lipopolysaccharide-activated monocytes from cord blood versus adult peripheral blood. AB - Monocytes (Mo) are critically important in the generation of inflammatory mediators, cytokines/chemokines, and regulation of innate and adaptive immune responses. We and others have previously demonstrated significant dysregulated cytokine gene expression and protein production and in vitro functional activities of activated cord blood (CB) vs adult peripheral blood (APB) mononuclear cells (MNC). In this study, we compared, by oligonucleotide microarray, the differential gene expression profiles of basal and LPS-activated APB vs CB Mo. We demonstrated a significant increase in the gene expression of several important functional groups of CB genes compared with basal levels including cytokine (IL-12p40, 5-fold), immunoregulatory (signaling lymphocytic activation molecule, 4-fold), signal transduction (Pim-2, 3-fold), and cell structure (Rho7, 4-fold) among others. Furthermore, there was significantly differentially amplified gene expression in LPS-activated APB vs LPS-activated CB Mo, including cytokine (G-CSF, 14-fold), chemokine (macrophage-inflammatory protein 1alpha, 5-fold), immunoregulatory (MHC DRB1, 5-fold), transcription factor (JunB, 4-fold), signal transduction (STAT4, 5-fold), apoptotic regulation (BAX, 5-fold), and cell structure (ladinin 1, 6-fold) among others. These results provide insight into the molecular basis for normal genetic regulation of Mo development and cellular function and differential inflammatory and innate and adaptive immune responses between activated CB and APB Mo. PMID- 15128767 TI - CD28 signals in the immature immunological synapse. AB - T cell recognition of peptide-MHC complexes on APCs results in the aggregation of TCRs at a central supramolecular activation complex (c-SMAC) within a mature immunological synapse. T cells require a second "costimulatory" signal for activation, the most important of which, for naive T cells, is from CD28. However the time at which CD28-derived signals are induced relative to c-SMAC formation is not well understood. In this study, we have assessed the kinetics of CD28 localization and function relative to well-established aspects of c-SMAC formation. CD28 accumulates at the immature synapse alongside the TCR and is likewise enriched at the synapse at the onset of the calcium signal. In addition, using CD28 deficient or reconstituted murine cells in a single-cell recording approach shows that CD28 regulates this signal within seconds of a TCR-mediated rise in intracellular calcium levels. Finally, CD28 exerts effects on both the initiation and stabilization of the synapse in parallel with its effects on the downstream proliferation of T cells. Together, the data show that CD28 functions in the immunological synapse before the formation of the c-SMAC. PMID- 15128768 TI - CD4 raft association and signaling regulate molecular clustering at the immunological synapse site. AB - T cell activation is associated with the partitioning of TCRs and other signaling proteins, forming an immunological synapse. This study demonstrates a novel function for the CD4 coreceptor in regulating molecular clustering at the immunological synapse site. We show using transgenic mouse and retroviral reconstitution studies that CD4 is required for TCR/protein kinase C (PKC) theta clustering. Specifically, we demonstrate that CD4 palmitoylation sequences are required for TCR/PKCtheta raft association and subsequent clustering, indicating a particular role for raft-associated CD4 molecules in regulating immune synapse organization. Although raft association of CD4 is necessary, it is not sufficient to mediate clustering, as cytoplasmic tail deletion mutants are able to localize to rafts, but are unable to mediate TCR/PKCtheta clustering, indicating an additional requirement for CD4 signaling. These studies suggest that CD4 coreceptor function is regulated not only through its known signaling function, but also by posttranslational lipid modifications which regulate localization of CD4 in lipid rafts. PMID- 15128769 TI - Depletion of CD8+ cells abolishes the pregnancy protective effect of progesterone substitution with dydrogesterone in mice by altering the Th1/Th2 cytokine profile. AB - One of the most remarkable immunological regulations is the maternal immune tolerance toward the fetal semiallograft during pregnancy, which has been referred to as immunity's pregnant pause. Rejection of the semiallogeneic trophoblast cells must be selectively inhibited and pathways presumably include Th2 cytokines unopposed by Th1 cytokines. Steroid hormones, including progesterone, have similar effects. Low levels of progesterone and Th2 cytokines and high levels of Th1 cytokines are attributable for increased abortions in mammalians, which may be triggered by psychoemotional stress. Thus, the aim of the present study was to provide experimental evidence for the mechanism involved in the mediation of immune responses by endocrine signals during pregnancy and stress-triggered pregnancy failure. DBA/2J-mated CBA/J female mice were randomized in three groups: 1) control females, 2) mice exposed to stress on gestation day 5.5, and 3) mice exposed to stress and substituted with dydrogesterone, a progestogen with a binding profile highly selective for the progesterone receptor on gestation day 5.5. On gestation days 7.5, 9.5, and 10.5, mice of each group were sacrificed, and the frequency of CD8(+) cells and cytokine expression (IL-4, IL-12, TNF-alpha, IFN-gamma) in blood and uterus cells was evaluated by flow cytometry. Additionally, some mice were depleted of CD8 cells by injection of mAb. We observed that progesterone substitution abrogated the abortogenic effects of stress exposure by decreasing the frequency of abortogenic cytokines. This pathway was exceedingly CD8-dependent, because depletion of CD8 led to a termination of the pregnancy protective effect of progesterone substitution. PMID- 15128770 TI - Generation of anergic and regulatory T cells following prolonged exposure to a harmless antigen. AB - Regulatory CD4(+) T cells are known to develop during the induction of donor specific peripheral tolerance to transplanted tissues; it is proposed that such tolerance is a consequence of persistent, danger-free stimulation by Ag. To test this hypothesis, male RAG-1(-/-) mice were recolonized with small numbers of monospecific CD4(+) T cells specific for the male H-2E(k)-restricted Ag Dby. After 6 wk in the male environment, the monospecific CD4(+) T cells, having recolonized the host, had become anergic to stimulation in vitro and had acquired a regulatory capacity. CD4(+) T cells in these mice expressed higher levels of CTLA-4 and glucocorticoid-induced TNF-related receptor than naive CD4(+) T cells, but only 3% of the recolonizing cells were CD25(+) and did not express significant foxP3 mRNA. In vivo, these tolerant T cells could censor accumulation of, and IFN-gamma production by, naive T cells, with only a slight inhibition of proliferation. This suppressive effect was not reversed by the addition of fresh bone marrow-derived male dendritic cells. These results suggest that persistent exposure to Ag in conditions that fail to evoke proinflammatory stimuli leads to the development of T cells that are both anergic and regulatory. PMID- 15128771 TI - Up-regulation of CD1d expression restores the immunoregulatory function of NKT cells and prevents autoimmune diabetes in nonobese diabetic mice. AB - The immunoregulatory function of NKT cells is crucial for prevention of autoimmunity. The prototypical NKT cell Ag alpha-galactosylceramide is not present in mammalian cells, and little is known about the mechanism responsible for NKT cell recruitment and activation. Up-regulation of CD1d, the NKT cell restriction molecule, expressed on mononuclear cells infiltrating the target organ, could represent the physiological trigger for NKT cells to self-contain T cell immunity and to prevent autoimmune disease. Recognition of CD1d, either by itself or bound to self-ligands (selfCD1d), could drive NKT cells toward an immunoregulatory phenotype. Hence, ineffective NKT cell-mediated immunoregulation in autoimmune-prone individuals including nonobese diabetic (NOD) mice could be related to defective signals that regulate CD1d expression at time and site of autoimmunity. To test this hypothesis, we transgenically overexpressed CD1d molecules under the control of the insulin promoter within the pancreatic islets of NOD mice (insCD1d). Recognition of overexpressed CD1d molecules rescued NKT cell immunoregulatory function and prevented autoimmune diabetes in insCD1d transgenic NOD mice. Protection from diabetes was associated with a biased IL-4 secreting cytokine phenotype of NKT cells and alteration of the cytokine microenvironment in the pancreatic lymph nodes of transgenic mice. The net effect was a reduced development of the autoimmune T cell repertoire. Our findings suggest that up-regulation of CD1d expression during inflammation is critical to maintain T cell homeostasis and to prevent autoimmunity. PMID- 15128772 TI - The inducible costimulator plays the major costimulatory role in humoral immune responses in the absence of CD28. AB - CD28 plays crucial costimulatory roles in T cell proliferation, cytokine production, and germinal center response. Mice that are deficient in the inducible costimulator (ICOS) also have defects in cytokine production and germinal center response. Because the full induction of ICOS in activated T cells depends on CD28 signal, the T cell costimulatory capacity of ICOS in the absence of CD28 has remained unclear. We have clarified this issue by comparing humoral immune responses in wild-type, CD28 knockout (CD28 KO), and CD28-ICOS double knockout (DKO) mice. DKO mice had profound defects in Ab responses against environmental Ags, T-dependent protein Ags, and vesicular stomatitis virus that extended far beyond those observed in CD28 KO mice. However, DKO mice mounted normal Ab responses against a T-independent Ag, indicating that B cell function itself was normal. Restimulated CD4(+) DKO T cells that had been primed in vivo showed decreased proliferation and reduced IL-4 and IL-10 production compared with restimulated CD4(+) T cells from CD28 KO mice. Thus, in the absence of CD28, ICOS assumes the major T cell costimulatory role for humoral immune responses. Importantly, CD28-mediated ICOS up-regulation is not essential for ICOS function in vivo. PMID- 15128773 TI - Heart, but not skin, allografts from donors lacking Flt3 ligand exhibit markedly prolonged survival time. AB - Fms-like tyrosine kinase 3 ligand (Flt3L) administration leads to dramatic increases in dendritic cells (DC) in lymphoid and nonlymphoid tissues. Conversely, mice lacking Flt3L (Flt3L(-)/(-)) show severe reductions in both myeloid (CD11c(+)CD8alpha(-)) and lymphoid-related DC (CD11c(+)CD8alpha(+)) in the thymus and secondary lymphoid organs. In this study marked reductions in CD11c(+) interstitial cardiac DC and in dermal, but not epidermal, DC (Langerhans cells) were also observed. CD11c(+) cells that migrated from Flt3L(-/-) skin explants expressed lower surface MHC class II and costimulatory molecules and naive T cell allostimulatory activity than migratory wild-type (wt) C57BL/6 (B6) CD11c(+) cells. We examined the survival of Flt3L(-)/(-) heart or tail skin grafts (H2(b)) in allogeneic wt (BALB/c; H2(d)) recipients. The outcome of transplantation of BALB/c organs into Flt3L(-)/(-) recipients was also determined. Flt3L(-)/(-) mice rejected BALB/c heart or skin grafts with similar kinetics as B6 wt recipients. Trafficking of donor DC into host spleens or draining lymph nodes was markedly reduced after transplantation of Flt3L(-)/(-) heart, but not skin grafts, respectively. Compared with wt hearts, survival of Flt3L(-)/(-) hearts was markedly prolonged in BALB/c recipients (median survival time, 37 and 15 days, respectively; p < 0.001). Skin graft survival was unaffected. Rejection of Flt3L(-/-) hearts was precipitated by infusion of wt donor DC at the time of transplant. Thus, severe depletion of interstitial heart DC resulting from targeted gene disruption prolongs, but does not indefinitely extend, heart survival. Acute rejection of wt grafts in Flt3L(-/-) recipients reflects presumably an intact role of the direct pathway of allorecognition. PMID- 15128774 TI - An inhibitory Ig superfamily protein expressed by lymphocytes and APCs is also an early marker of thymocyte positive selection. AB - Positive selection of developing thymocytes is associated with changes in cell function, at least in part caused by alterations in expression of cell surface proteins. Surprisingly, however, few such proteins have been identified. We have analyzed the pattern of gene expression during the early stages of murine thymocyte differentiation. These studies led to identification of a cell surface protein that is a useful marker of positive selection and is a likely regulator of mature lymphocyte and APC function. The protein is a member of the Ig superfamily and contains conserved tyrosine-based signaling motifs. The gene encoding this protein was independently isolated recently and termed B and T lymphocyte attenuator (Btla). We describe in this study anti-BTLA mAbs that demonstrate that the protein is expressed in the bone marrow and thymus on developing B and T cells, respectively. BTLA is also expressed by all mature lymphocytes, splenic macrophages, and mature, but not immature bone marrow derived dendritic cells. Although mice deficient in BTLA do not show lymphocyte developmental defects, T cells from these animals are hyperresponsive to anti-CD3 Ab stimulation. Conversely, anti-BTLA Ab can inhibit T cell activation. These results implicate BTLA as a negative regulator of the activation and/or function of various hemopoietic cell types. PMID- 15128775 TI - Curcumin (diferuloylmethane) inhibits receptor activator of NF-kappa B ligand induced NF-kappa B activation in osteoclast precursors and suppresses osteoclastogenesis. AB - Numerous studies have indicated that inflammatory cytokines play a major role in osteoclastogenesis, leading to the bone resorption that is frequently associated with cancers and other diseases. Gene deletion studies have shown that receptor activator of NF-kappaB ligand (RANKL) is one of the critical mediators of osteoclastogenesis. How RANKL mediates osteoclastogenesis is not fully understood, but an agent that suppresses RANKL signaling has potential to inhibit osteoclastogenesis. In this report, we examine the ability of curcumin (diferuloylmethane), a pigment derived from turmeric, to suppress RANKL signaling and osteoclastogenesis in RAW 264.7 cells, a murine monocytic cell line. Treatment of these cells with RANKL activated NF-kappaB, and preexposure of the cells to curcumin completely suppressed RANKL-induced NF-kappaB activation. Curcumin inhibited the pathway leading from activation of IkappaBalpha kinase and IkappaBalpha phosphorylation to IkappaBalpha degradation. RANKL induced osteoclastogenesis in these monocytic cells, and curcumin inhibited both RANKL- and TNF-induced osteoclastogenesis and pit formation. Curcumin suppressed osteoclastogenesis maximally when added together with RANKL and minimally when it was added 2 days after RANKL. Whether curcumin inhibits RANKL-induced osteoclastogenesis through suppression of NF-kappaB was also confirmed independently, as RANKL failed to activate NF-kappaB in cells stably transfected with a dominant-negative form of IkappaBalpha and concurrently failed to induce osteoclastogenesis. Thus overall these results indicate that RANKL induces osteoclastogenesis through the activation of NF-kappaB, and treatment with curcumin inhibits both the NF-kappaB activation and osteoclastogenesis induced by RANKL. PMID- 15128776 TI - Conversion of CTLA-4 from inhibitor to activator of T cells with a bispecific tandem single-chain Fv ligand. AB - Abs or their recombinant fragments against surface receptors of the Ig superfamily can induce or block the receptors' native function depending on whether they induce or prevent the assembly of signalosomes on their cytoplasmic tails. In this study, we introduce a novel paradigm based on the observation that a bispecific tandem single-chain variable region fragment ligand of CTLA-4 by itself converts this inhibitory receptor into an activating receptor for primary human T lymphocytes. This reversal of function results from increased recruitment of the serine/threonine phosphatase 2A to the cytoplasmic tail of CTLA-4, consistent with a role of this phosphatase in the regulation of CTLA-4 function, and assembly of a distinct signalosome that activates an lck-dependent signaling cascade and induces IL-2 production. Our data demonstrate that the cytoplasmic domain of CTLA-4 has an inherent plasticity for signaling that can be exploited therapeutically with recombinant ligands for this receptor. PMID- 15128777 TI - IL-4 confers NK stimulatory capacity to murine dendritic cells: a signaling pathway involving KARAP/DAP12-triggering receptor expressed on myeloid cell 2 molecules. AB - Dendritic cells (DC) regulate NK cell functions, but the signals required for the DC-mediated NK cell activation, i.e., DC-activated NK cell (DAK) activity, remain poorly understood. Upon acute inflammation mimicked by LPS or TNF-alpha, DC undergo a maturation process allowing T and NK cell activation in vitro. Chronic inflammation is controlled in part by Th2 cytokines. In this study, we show that IL-4 selectively confers to DC NK but not T cell stimulatory capacity. IL-4 is mandatory for mouse bone marrow-derived DC grown in GM-CSF (DC(GM/IL-4)) to promote NK cell activation in the draining lymph nodes. IL-4-mediated DAK activity depends on the KARAP/DAP12-triggering receptor expressed on myeloid cell 2 signaling pathway because: 1) gene targeting of the adaptor molecule KARAP/DAP12, a transmembrane polypeptide with an intracytoplasmic immunoreceptor tyrosine-based activation motif, suppresses the DC(GM/IL-4) capacity to activate NK cells, and 2) IL-4-mediated DAK activity is significantly blocked by soluble triggering receptor expressed on myeloid cell 2 Fc molecules. These data outline a novel role for Th2 cytokines in the regulation of innate immune responses through triggering receptors expressed on myeloid cells. PMID- 15128778 TI - Autoreactive T cells in healthy individuals. AB - The presence of autoreactive CD4(+) T cells in the peripheral blood of healthy human subjects was investigated after removal of CD4(+)CD25(+) regulatory T cells (Treg). CD4(+) T cells that were directed against the type 1 diabetes-associated autoantigen glutamic acid decarboxylase 65, the melanocyte differentiation Ag tyrosinase, and the cancer/testis tumor Ag NY-ESO-1 were readily derived from PBMC of healthy individuals. These autoreactive T cells could be visualized, using Ag-specific class II tetramer reagents, in the peripheral blood of most individuals examined. Addition of CD4(+)CD25(+) Treg back to the CD4(+)CD25(-) population suppressed the expansion of the autoreactive T cells. Autoreactive T cells were cloned based on tetramer binding, and expressed characteristic activation markers upon self-Ag stimulation. These results show that autoreactive T cells are present in most healthy individuals and that Treg likely play an important role of keeping these autoreactive T cells in check. PMID- 15128779 TI - IL-2 induces a competitive survival advantage in T lymphocytes. AB - The acquisition of long-term survival potential by activated T lymphocytes is essential to ensure the successful development of a memory population in the competitive environment of the lymphoid system. The factors that grant competitiveness for survival to primed T cells are poorly defined. We examined the role of IL-2 signals during priming of CD4(+) T cells in the induction of a long-lasting survival program. We show that Ag-induced cycling of CD4(+) IL-2(-/ ) T cells is independent of IL-2 in vitro. However, IL-2(-/-) T cells failed to accumulate in large numbers and develop in effector cells when primed in the absence of IL-2. More importantly, Ag-activated IL-2(-/-) T cells were unable to survive for prolonged periods of time after adoptive transfer in unmanipulated, syngeneic mice. IL-2(-/-) T cells exposed to IL-2 signals during priming, however, acquired a robust and long-lasting survival advantage over cells that cycled in the absence of IL-2. Interestingly, this IL-2-induced survival program was required for long-term persistence of primed IL-2(-/-) T cells in an intact lymphoid compartment, but was unnecessary in a lymphopenic environment. Therefore, IL-2 enhances competitiveness for survival in CD4(+) T cells, thereby facilitating the development of a memory population. PMID- 15128780 TI - Adult human liver contains CD8pos T cells with naive phenotype, but is not a site for conventional alpha beta T cell development. AB - Normal adult human liver (AHL) contains populations of unconventional lymphocytes that have been shown in the mouse to mature locally. The presence of lymphoid progenitors together with IL-7, recombinase-activating gene, and pre-TCR-alpha expression in AHL suggests similar local T cell development activity in humans. Flow cytometry was used to characterize potentially naive hepatic alphabeta-T cells. We looked for evidence of TCR-alphabeta cell development in AHL by quantifying delta deletion TCR excision circles (TRECs) in CD3(pos) populations isolated from the liver and matched blood of eight individuals. Phenotypic analysis of hepatic T cells suggests the presence of Ag-inexperienced populations. TRECs were detected in all blood samples (mean, 164.10 TRECs/ micro g DNA), whereas only two hepatic samples were positive at low levels (59.40 and 1.92). The relatively high level of CD8(pos) T cells in these livers with a naive phenotype suggests that in addition to its role as a graveyard for Ag-specific activated CD8(pos) T cells, naive CD8(pos) T cells may enter the liver without prior activation. The almost complete absence of TRECs suggests that normal AHL is not a site for the development of conventional alphabeta T cells. PMID- 15128781 TI - IL-10-secreting regulatory T cells do not express Foxp3 but have comparable regulatory function to naturally occurring CD4+CD25+ regulatory T cells. AB - Regulatory T cells (T(Reg)) control immune responses to self and nonself Ags. The relationship between Ag-driven IL-10-secreting T(Reg) (IL-10-T(Reg)) and naturally occurring CD4(+)CD25(+) T(Reg) is as yet unclear. We show that mouse IL 10-T(Reg) obtained using either in vitro or in vivo regimens of antigenic stimulation did not express the CD4(+)CD25(+) T(Reg)-associated transcription factor Foxp3. However, despite the absence of Foxp3 expression, homogeneous populations of IL-10-T(Reg) inhibited the in vitro proliferation of CD4(+)CD25(-) T cells with a similar efficiency to that of CD4(+)CD25(+) T(Reg). This inhibition of T cell proliferation by IL-10-T(Reg) was achieved through an IL-10 independent mechanism as seen for CD4(+)CD25(+) T(Reg) and was overcome by exogenous IL-2. Both IL-10-T(Reg) and CD4(+)CD25(+) T(Reg) were similar in that they produced little to no IL-2. These data show that Foxp3 expression is not a prerequisite for IL-10-T(Reg) activity in vitro or in vivo, and suggest that IL 10-T(Reg) and naturally occurring CD4(+)CD25(+) T(Reg) may have distinct origins. PMID- 15128782 TI - Thymic expression of a gastritogenic epitope results in positive selection of self-reactive pathogenic T cells. AB - Intrathymic expression of tissue-specific self-Ags can mediate tolerance of self reactive T cells. However, in this study we define circumstances by which thymic expression of a tissue-specific autoepitope enhances positive selection of disease-causing, self-reactive T cells. An immunodominant gastritogenic epitope, namely the gastric H/K ATPase beta subunit(253-277) (H/Kbeta(253-277)), was attached to the C terminus of the invariant chain (Ii) and the hybrid Ii (Ii H/Kbeta(253-277)) expressed in mice under control of the Ii promoter. The Ii H/Kbeta(253-277) fusion protein was localized to MHC class II-expressing cells in the thymus and periphery of Ii-H/Kbeta(253-277) transgenic mice. In one transgenic line the level of presentation in the periphery (spleen) was insufficient to activate naive, low affinity H/Kbeta(253-277)-specific transgenic T cells (1E4-TCR), whereas thymic presentation of H/Kbeta(253-277) enhanced positive selection of 1E4-TCR cells in Ii-H/Kbeta(253-277)/1E4-TCR double transgenic mice. Furthermore, Ii-H/Kbeta(253-277)/1E4-TCR double-transgenic mice had an increased incidence of autoimmune gastritis compared with 1E4-TCR single transgenic mice, demonstrating that the 1E4 T cells that seeded the periphery of Ii-H/Kbeta(253-277) mice were pathogenic. Therefore, low levels of tissue specific Ags in the thymus can result in positive selection of low avidity, self reactive T cells. These findings also suggest that the precise level of tissue specific Ags in the thymus may be an important consideration in protection against autoimmune disease and that perturbation of the levels of self-Ags may be detrimental. PMID- 15128783 TI - Induction of foxP3+ regulatory T cells in the periphery of T cell receptor transgenic mice tolerized to transplants. AB - Transplantation tolerance can be induced in mice by grafting under the cover of nondepleting CD4 plus CD8 or CD154 mAbs. This tolerance is donor Ag specific and depends on a population of CD4(+) regulatory T cells that, as yet, remain poorly defined in terms of their specificity, origin, and phenotype. Blocking of the Ag specific response in vitro with an anti-CD4 mAb allowed T cells from monospecific female TCR-transgenic mice against the male Ag Dby, presented by H-2E(k), to express high levels of foxP3 mRNA. foxP3 induction was dependent on TGF-beta. The nondepleting anti-CD4 mAb was also able to induce tolerance in vivo in such monospecific TCR-transgenic mice, and this too was dependent on TGF-beta. As in conventional mice, acquired tolerance was dominant, such that naive monospecific T cells were not able to override tolerance. Splenic T cells from tolerant mice proliferated normally in response to Ag, and secreted IFN-gamma and some IL-4, similar to control mice undergoing primary or secondary graft rejection. High levels of foxP3 mRNA, and glucocorticoid-induced TNFR superfamily member 18 (GITR)(+) CD25(+) T cells were found within the tolerated skin grafts of long term tolerant recipients. These data suggest that regulatory T cells maintaining transplantation tolerance after CD4 Ab blockade can be induced de novo through a TGF-beta-dependent mechanism, and come to accumulate in tolerated grafts. PMID- 15128784 TI - The serotoninergic receptors of human dendritic cells: identification and coupling to cytokine release. AB - The neurotransmitter 5-hydroxytryptamine (5-HT), commonly known as serotonin, is stored at peripheral sites in mast cells and released from this peripheral source upon IgE cross-linking. In this study, we investigated the expression of serotoninergic receptors (5-HTR), the signaling pathway, and biological activity of 5-HT on human dendritic cells (DC), showing that immature and mature DC expressed mRNA for different serotoninergic receptors. Thereby, the mRNA of 5 HTR(1B), 5-HTR(1E), 5-HTR(2A), 5-HTR(2B), one splicing variant of the 5-HTR(3), 5 HTR(4), and 5-HTR(7) receptors were detected. Immature DC preferentially expressed mRNA for the heptahelical 5-HTR(1B), 5-HTR(1E), and 5-HTR(2B) receptors, while mature DC mostly expressed 5-HTR(4) and 5-HTR(7). The mRNA expression level of the ligand-gated cation channel 5-HTR(3) and the heptahelical 5-HTR(2A) did not significantly change during maturation. Isotype-selective receptor agonists allowed us to show that 5-HT stimulated 5-HTR(3)-dependent Ca(2+) influx in immature and mature DC. Moreover, we revealed that 5-HTR(1) and 5-HTR(2) receptor stimulation induced intracellular Ca(2+) mobilization via G(i/o) proteins in immature, but not mature, DC. Activation of 5-HTR(4) and 5 HTR(7) induced cAMP elevation in mature DC. Functional studies indicated that activation of 5-HTR(4) and 5-HTR(7) enhanced the release of the cytokines IL 1beta and IL-8, while reducing the secretion of IL-12 and TNF-alpha in mature DC. In summary, our study shows that 5-HT stimulated, in a maturation-dependent manner, different signaling pathways in DC. These data point to a role for 5-HT in regulating the immune response at peripheral sites. PMID- 15128785 TI - The role of IL-5 for mature B-1 cells in homeostatic proliferation, cell survival, and Ig production. AB - B-1 cells, distinguishable from conventional B-2 cells by their cell surface marker, anatomical location, and self-replenishing activity, play an important role in innate immune responses. B-1 cells constitutively express the IL-5R alpha chain (IL-5Ralpha) and give rise to Ab-producing cells in response to various stimuli, including IL-5 and LPS. Here we report that the IL-5/IL-5R system plays an important role in maintaining the number and the cell size as well as the functions of mature B-1 cells. The administration of anti-IL-5 mAb into wild-type mice, T cell-depleted mice, or mast cell-depleted mice resulted in reduction in the total number and cell size of B-1 cells to an extent similar to that of IL 5Ralpha-deficient (IL-5Ralpha(-/-)) mice. Cell transfer experiments have demonstrated that B-1 cell survival in wild-type mice and homeostatic proliferation in recombination-activating gene 2-deficient mice are impaired in the absence of IL-5Ralpha. IL-5 stimulation of wild-type B-1 cells, but not IL 5Ralpha(-/-) B-1 cells, enhances CD40 expression and augments IgM and IgG production after stimulation with anti-CD40 mAb. Enhanced IgA production in feces induced by the oral administration of LPS was not observed in IL-5Ralpha(-/-) mice. Our results illuminate the role of IL-5 in the homeostatic proliferation and survival of mature B-1 cells and in IgA production in the mucosal tissues. PMID- 15128786 TI - The ability of two Listeria monocytogenes vaccines targeting human papillomavirus 16 E7 to induce an antitumor response correlates with myeloid dendritic cell function. AB - Previous work from our laboratory has shown that Lm-LLO-E7 induces complete regression of approximately 75% of established TC-1 tumors, whereas Lm-E7 only slows the growth of such tumors. In this study, we examine the effects of Lm-LLO E7 vs Lm-E7 on APCs. We hypothesize that the difference in antitumor efficacy of the two vaccines is due to the ability of each of these vectors to render immature dendritic cells (DCs) effective APCs in terms of MHC class II or costimulatory molecule expression. We also examine the ability of these vectors to stimulate cytokine production by DCs. Both vectors induced IL-12 and TNF alpha, but only Lm-LLO-E7 induced IL-2 production by DCs. Lm-LLO-E7 also induced significantly higher levels of MHC class II molecules, CD40, and B7 costimulatory molecules (CD86, B7-H1, and B7-DC) on DCs than Lm-E7. Interestingly, a shift of CD11c(+) cells from CD86(low) to CD86(high) is observed post-Lm-LLO-E7 infection. A similar shift is also observed for B7-H1 and B7-DC molecules. Moreover, Lm-LLO E7, but not Lm-E7-pulsed DCs, stimulate naive T cell proliferation. These results indicate that Lm-LLO-E7 is more effective than Lm-E7 at inducing DC maturation. This effect is independent of the E7 Ag, because Lm-LLO-NP, and a mixture of Lm LLO-NP and Lm-E7 induce the same changes in DC phenotype as Lm-LLO-E7. Taken together, the changes in DC expression correlate well with the differences in antitumor efficacy between these two vaccines. PMID- 15128787 TI - Stimulation by soluble CD70 promotes strong primary and secondary CD8+ cytotoxic T cell responses in vivo. AB - Identification of the signals required for optimal differentiation of naive CD8(+) T cells into effector and memory cells is critical for the design of effective vaccines. In this study we demonstrate that CD27 stimulation by soluble CD70 considerably enhances the magnitude and quality of the CD8(+) T cell response. Stimulation with soluble CD70 in the presence of Ag significantly enhanced the proliferation of CD8(+) T cells and their ability to produce IL-2 and IFN-gamma in vitro. Administration of Ag and soluble CD70 resulted in a massive (>300-fold) expansion of Ag-specific CD8(+) T cells in vivo, which was due to the enhanced proliferation and survival of activated T cells. In mice that received Ag and soluble CD70, CD8(+) T cells developed into effectors with direct ex vivo cytotoxicity. Furthermore, unlike peptide immunization, which resulted in a diminished response after rechallenge, CD27 stimulation during the primary challenge evoked a strong secondary response upon rechallenge with the antigenic peptide. Thus, in addition to increasing the frequency of primed Ag-specific T cells, CD27 signaling during the primary response instills a program of differentiation that allows CD8(+) T cells to overcome a state of unresponsiveness. Taken together these results demonstrate that soluble CD70 has potent in vivo adjuvant effects for CD8(+) T cell responses. PMID- 15128788 TI - The role of the p38 mitogen-activated protein kinase, extracellular signal regulated kinase, and phosphoinositide-3-OH kinase signal transduction pathways in CD40 ligand-induced dendritic cell activation and expansion of virus-specific CD8+ T cell memory responses. AB - Mature dendritic cells (DCs) are central to the development of optimal T cell immune responses. CD40 ligand (CD40L, CD154) is one of the most potent maturation stimuli for immature DCs. We studied the role of three signaling pathways, p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), and phosphoinositide-3-OH kinase (PI3K), in CD40L-induced monocyte-derived DC activation, survival, and expansion of virus-specific CD8(+) T cell responses. p38 MAPK pathway was critical for CD40L-mediated up-regulation of CD83, a marker of DC maturation. CD40L-induced monocyte-derived DC IL-12 production was mediated by both the p38 MAPK and PI3K pathways. CD40L-mediated DC survival was mostly mediated by the PI3K pathway, with smaller contributions by p38 MAPK and ERK pathways. Finally, the p38 MAPK pathway was most important in mediating CD40L stimulated DCs to induce strong allogeneic responses as well as expanding virus specific memory CD8(+) T cell responses. Thus, although the p38 MAPK, PI3K, and ERK pathways independently affect various parameters of DC maturation induced by CD40L, the p38 MAPK pathway within CD40L-conditioned DCs is the most important pathway to maximally elicit T cell immune responses. This pathway should be exploited in vivo to either completely suppress or enhance CD8(+) T cell immune responses. PMID- 15128789 TI - T cells associated with tumor regression recognize frameshifted products of the CDKN2A tumor suppressor gene locus and a mutated HLA class I gene product. AB - The dramatic tumor regression observed following adoptive T cell transfer in some patients has led to attempts to identify novel Ags to understand the nature of these responses. Nearly complete regression of multiple metastatic melanoma lesions was observed in patient 1913 following adoptive transfer of autologous tumor-infiltrating lymphocytes. The autologous 1913 melanoma cell line expressed a mutated HLA-A11 class I gene product that was recognized by the bulk tumor infiltrating lymphocytes as well as a dominant T cell clone derived from this line. A second dominant T cell clone, T1D1, did not recognize the mutated HLA-A11 product, but recognized an allogeneic melanoma cell line that shared expression of HLA-A11 with the parental tumor cell line. Screening of an autologous melanoma cDNA library with clone T1D1 T cells in a cell line expressing the mutated HLA A11 gene product resulted in the isolation of a p14ARF transcript containing a 2 bp deletion in exon 2. The T cell epitope recognized by T1D1, which was encoded within the frameshifted region of the deleted p14ARF transcript, was also generated from frameshifted p14ARF or p16INK4a transcripts that were isolated from two additional melanoma cell lines. The results of monitoring studies indicated that T cell clones reactive with the mutated HLA-A11 gene product and the mutated p14ARF product were highly represented in the peripheral blood of patient 1913 1 wk following adoptive transfer, indicating that they may have played a role in the nearly complete tumor regression that was observed following this treatment. PMID- 15128791 TI - The tumoricidal activity of memory CD8+ T cells is hampered by persistent systemic antigen, but full functional capacity is regained in an antigen-free environment. AB - Naive T cells can be tolerized in the periphery by diverse mechanisms. However, the extent to which memory T cells are susceptible to tolerance induction is less well defined. Vaccination of mice with a minimal CTL epitope derived from human adenovirus type 5 E1A in IFA s.c. readily tolerizes naive as well as recently activated CD8(+) T cells due to the overwhelming systemic and persistent presence of the peptide. We have now studied the effect of this peptide on established memory cells, which were induced at least 50 days before by virus vaccination. Memory cells did not undergo peripheral deletion and kept their ability to produce IFN-gamma as well as their cytolytic activity in response to Ag directly ex vivo. However, memory CTL responses in virus vaccinated mice injected with peptide ceased to control tumor outgrowth. Interestingly, functional capacities were regained when T cells were transferred to an Ag-free environment in vivo as determined by their ability to reject an otherwise lethal tumor challenge. Together, these findings indicate that memory CTL responses can be functionally incapacitated, but are not, in contrast to naive or recently activated T cells, irreversibly tolerized by persistent systemic Ag, as memory T cells quickly regain effector function upon disappearance of the Ag. PMID- 15128790 TI - Toll-like receptor ligands directly promote activated CD4+ T cell survival. AB - Toll-like receptor (TLR) engagement by pathogen-associated molecular patterns (PAMPs) is an important mechanism for optimal cellular immune responses. APC TLR engagement indirectly enhances activated CD4(+) T cell proliferation, differentiation, and survival by promoting the up-regulation of costimulatory molecules and the secretion of proinflammatory cytokines. However, TLRs are also expressed on CD4(+) T cells, suggesting that PAMPs may also act directly on activated CD4(+) T cells to mediate functional responses. In this study, we show that activated mouse CD4(+) T cells express TLR-3 and TLR-9 but not TLR-2 and TLR 4. Treatment of highly purified activated CD4(+) T cells with the dsRNA synthetic analog poly(I:C) and CpG oligodeoxynucleotides (CpG DNA), respective ligands for TLR-3 and TLR-9, directly enhanced their survival without augmenting proliferation. In contrast, peptidoglycan and LPS, respective ligands for TLR-2 and TLR-4 had no effect. Enhanced survival mediated by either poly(I:C) or CpG DNA required NF-kappaB activation and was associated with Bcl-x(L) up-regulation. However, only CpG DNA, but not poly(I:C)-mediated effects on activated CD4(+) T cells required the TLR/IL-1R domain containing adaptor molecule myeloid differentiation factor 88. Collectively, our results demonstrate that PAMPs can directly promote activated CD4(+) T cell survival, suggesting that TLRs on T cells can directly modulate adaptive immune responses. PMID- 15128792 TI - The Toll-like receptor-2/6 agonist macrophage-activating lipopeptide-2 cooperates with IFN-gamma to reverse the Th2 skew in an in vitro allergy model. AB - Dendritic cells (DC) are the most potent APCs with the capacity to induce, modulate, or shut down immune function. These features make them potentially useful for treating diseases associated with misled immunologic responses. Therefore, it was the aim of this study to reverse the allergen-dependent Th2 reaction responsible for allergic symptoms by modulating DC function. This issue was addressed in an in vitro test system consisting of human monocyte-derived allergen-pulsed DC from allergics cocultured with autologous lymphocytes. A Th2 reaction judged by the amplification of IL-4 and the down-regulation of IFN-gamma was induced by pulsing DC with the relevant allergen. To modulate this reaction, the Toll-like receptor 2/6 engaging mycoplasmal lipopetide macrophage-activating lipopeptide 2 kDa was combined with IFN-gamma to stimulate allergen-pulsed DC. Such treatment resulted in a 500-fold increase in IFN-gamma production in the supernatant of cocultured autologous lymphocytes, while the Th2 marker IL-4 was not affected. This phenomenon was associated with an increase in proliferation and the number of IFN-gamma-producing lymphocytes. Phenotype and function of thus treated DC remained stable. These data indicate that a former allergen-dependent Th2 reaction can be reversed toward a Th1-type response by an appropriate treatment of DC. PMID- 15128793 TI - Glycoprotein 96 can chaperone both MHC class I- and class II-restricted epitopes for in vivo presentation, but selectively primes CD8+ T cell effector function. AB - The ability of mature T lymphocytes to develop effector capacity after encounter with cognate Ag is generally dependent upon inflammatory signals associated with infection that induce dendritic cell activation/maturation. These inflammatory signals can derive directly from pathogens or can be expressed by host cells in response to infection. Heat shock proteins (HSPs) are a class of host-derived inflammatory mediators that perform the dual function of both chaperoning MHC class I-restricted epitopes into the cross-presentation pathway of DCs and inducing the activation/maturation of these DCs to allow priming of cognate CD8(+) T cell effector responses. Although the ability of HSPs to elicit effector CD8 cell responses has been well established, their potential to prime CD4 cell effector responses has been relatively unexplored. In the current study we compared the ability of the endoplasmic reticulum-resident HSP gp96 to prime CD4 vs CD8 cells using TCR transgenic adoptive transfer systems and soluble gp96 peptide complexes. As expected, gp96 facilitated the cross-presentation of a class I-restricted peptide and priming of effector function in cognate CD8 cells. Interestingly, gp96 also facilitated the in vivo presentation of a class II restricted peptide; however, the resulting CD4 cell response did not involve the development of effector function. Taken together, these data suggest that gp96 is an inflammatory mediator that selectively primes CD8 cell effector function. PMID- 15128794 TI - Restoration of NK T cell development in fyn-mutant mice by a TCR reveals a requirement for Fyn during early NK T cell ontogeny. AB - NK T cells are a unique lymphocyte population that have developmental requirements distinct from conventional T cells. Mice lacking the tyrosine kinase Fyn have 5- to 10-fold fewer mature NK T cells. This study shows that Fyn deficient mice have decreased numbers of NK1.1(-) NK T cell progenitors as well. 5-Bromo-2'-deoxyuridine-labeling studies indicate that the NK T cells remaining in fyn(-/-) mice exhibit a similar turnover rate as wild-type cells. The fyn(-/-) NK T cells respond to alpha-galactosylceramide, a ligand recognized by NK T cells, and produce cytokines, but have depressed proliferative capacity. Transgenic expression of the NK T cell-specific TCR alpha-chain Valpha14Jalpha18 leads to a complete restoration of NK T cell numbers in fyn(-/-) mice. Together, these results suggest that Fyn may have a role before alpha-chain rearrangement rather than for positive selection or the peripheral upkeep of cell number. NK T cells can activate other lymphoid lineages via cytokine secretion. These secondary responses are impaired in Fyn-deficient mice, but occur normally in fyn mutants expressing the Valpha14Jalpha18 transgene. Because this transgene restores NK T cell numbers, the lack of secondary lymphocyte activation in the fyn-mutant mice is due to the decreased numbers of NK T cells present in the mutant, rather than an intrinsic defect in the ability of the other fyn(-/-) lymphoid populations to respond. PMID- 15128795 TI - IL-9-induced expansion of B-1b cells restores numbers but not function of B-1 lymphocytes in xid mice. AB - Mice expressing the X-linked immunodeficiency (xid) mutation lack functional Bruton's tyrosine kinase and were shown to be specifically deficient in peritoneal B-1 lymphocytes. We have previously shown that IL-9, a cytokine produced by TH2 lymphocytes, promotes B-1 cell expansion in vivo. To determine whether IL-9 overexpression might compensate the xid mutation for B-1 lymphocyte development, we crossed xid mice with IL-9-transgenic mice. In this model, IL-9 restored normal numbers of mature peritoneal B-1 cells that all belonged to the CD5(-) B-1b subset. Despite this normal B-1 lymphocyte number, IL-9 failed to restore classical functions of B-1 cells, namely, the production of natural IgM Abs, the T15 Id Ab response to phosphorylcholine immunization, and the antipolysaccharide humoral response against Streptococcus pneumoniae. By using bromelain-treated RBC, we showed that the antigenic repertoire of these IL-9 induced B-1b lymphocytes was different from the repertoire of classical CD5(+) B 1a cells, indicating that the lack of B-1 function by B-1b cells is associated with distinct Ag specificities. Taken together, our data show that B-1b cell development can restore the peritoneal B-1 population in xid mice but that these B-1b cells are functionally distinct from CD5(+) B-1a lymphocytes. PMID- 15128796 TI - The size and phenotype of virus-specific T cell populations is determined by repetitive antigenic stimulation and environmental cytokines. AB - Based on the expression of the TNFR SFP CD27, two Ag-primed CD8(+) T cell subsets can be discerned in the circulation of healthy individuals: CD27(+) T cells that produce a variety of cytokines but do not display immediate cytolytic activity; and cytotoxic CD27(-) T cells, which secrete only IFN-gamma and TNF-alpha. The mechanism that controls the generation of these different phenotypes is unknown. We show that CMV reactivation not only increases the number of virus-specific T cells but also induces their transition from a CD27(+) to a CD27(-) phenotype. In support of a relation between pool size and phenotype in a cohort of latently infected individuals, the number of Ag-specific CD27(-) CD8(+) T cells was found to be linearly related to the total number of CMV-specific CD8(+) T cells. In vitro studies revealed that the acquisition of the CD27(-) phenotype on CMV specific T cells depended on the interaction of CD27 with its cellular ligand, CD70. Expression of CD70 was proportional to the amount of antigenic stimulation and blocked by the CD4(+) T cell-derived cytokine IL-21. Thus, induction of CD70, which may vary in distinct viral infections, appears to be a key factor in determining the size and phenotype of the CMV-specific T cell population in latently infected individuals. PMID- 15128797 TI - CD1d-independent NKT cells in beta 2-microglobulin-deficient mice have hybrid phenotype and function of NK and T cells. AB - Unlike CD1d-restricted NK1.1(+)TCRalphabeta(+) (NKT) cells, which have been extensively studied, little is known about CD1d-independent NKT cells. To characterize their functions, we analyzed NKT cells in beta(2)-microglobulin (beta(2)m)-deficient B6 mice. They are similar to NK cells and expressed NK cell receptors, including Ly49, CD94/NKG2, NKG2D, and 2B4. NKT cells were found in normal numbers in mice that are deficient in beta(2)m, MHC class II, or both. They were also found in the male HY Ag-specific TCR-transgenic mice independent of positive or negative selection in the thymus. For functional analysis of CD1d independent NKT cells, we developed a culture system in which CD1d-independent NKT cells, but not NK, T, or most CD1d-restricted NKT cells, grew in the presence of an intermediate dose of IL-2. IL-2-activated CD1d-independent NKT cells were similar to IL-2-activated NK cells and efficiently killed the TAP-mutant murine T lymphoma line RMA-S, but not the parental RMA cells. They also killed beta(2)m deficient Con A blasts, but not normal B6 Con A blasts, indicating that the cytotoxicity is inhibited by MHC class I on target cells. IL-2-activated NKT cells expressing transgenic TCR specific for the HY peptide presented by D(b) killed RMA-S, but not RMA, cells. They also killed RMA (H-2(b)) cells that were preincubated with the HY peptide. NKT cells from beta(2)m-deficient mice, upon CD3 cross-linking, secreted IFN-gamma and IL-2, but very little IL-4. Thus, CD1d independent NKT cells are significantly different from CD1d-restricted NKT cells. They have hybrid phenotypes and functions of NK cells and T cells. PMID- 15128798 TI - Human CD4+CD25+ regulatory T cells share equally complex and comparable repertoires with CD4+CD25- counterparts. AB - CD4(+)CD25(+) T cells are critical mediators of peripheral immune tolerance. However, many developmental and functional characteristics of these cells are unknown, and knowledge of human regulatory T cells is particularly limited. To better understand how human CD4(+)CD25(+) T cells develop and function, we examined the diversity of CD4(+)CD25(+) and CD4(+)CD25(-) T cell repertoires in both thymus and peripheral blood. Levels of T receptor excision circles (TREC) were comparable in purified CD4(+)CD25(+) and CD4(+)CD25(-) thymic populations, but were significantly higher than those in samples derived from peripheral blood, consistent with murine studies demonstrating thymic development of CD4(+)CD25(+) regulatory T cells. Surprisingly, CD4(+)CD25(-) T cells isolated from peripheral blood had greater TREC quantities than their CD4(+)CD25(+) counterparts, supporting the possibility of extrathymic expansion as well. CD4(+)CD25(+) and CD4(+)CD25(-) T cells from a given individual showed overlapping profiles with respect to diversity by Vbeta staining and spectratyping. Interestingly, CD4(+)CD25(+) T cells have lower quantities of CD3 than CD4(+)CD25(-) T cells. Collectively, these data suggest that human CD4(+)CD25(+) T cells recognize a similar array of Ags as CD4(+)CD25(-) T cells. However, reduced levels of TCR on regulatory T cells suggest different requirements for activation and may contribute to how the immune system regulates whether a particular response is suppressed or augmented. PMID- 15128799 TI - Loading of MHC class I and II presentation pathways by exogenous antigens: a quantitative in vivo comparison. AB - The MHC class I pathway is usually fueled by endogenous Ags, while exogenous Ags reach the MHC class II pathway. Although exogenous epitopes may also enter the MHC class I pathway, quantification of the efficiency of the process has remained a difficult task. In an attempt of such a quantification, we directly compared the amount of exogenous virus-like particles required for induction of cytotoxic T cell responses by cross-priming with the amount of virus-like particles required for induction of Th cell responses by the conventional route of MHC class II loading as an internal standard. Surprisingly, we found that cross presentation of peptides derived from exogenous Ags on MHC class I molecules is of only marginally lower efficiency ( approximately 1- to 10-fold) than the classical MHC class II pathway in vitro and in vivo. Thus, Ag quantities required for cross-presentation and cross-priming are similar to those required for fueling the MHC class II pathway. PMID- 15128800 TI - IL-2 secretion by CD4+ T cells in vivo is rapid, transient, and influenced by TCR specific competition. AB - The secretion of IL-2 is a critical and early landmark in the activation program of CD4(+) T cells in vitro, but the lack of sensitive assays has limited its application for studying T cell activation in vivo. Using a mouse cytokine capture assay we were able to detect the rapid secretion of IL-2 after an in vivo stimulus by 1-2 h in naive T cells and as early as 30 min in memory T cells. Maximal secretion was achieved within 1-2 h for memory cells or 6-8 h for naive T cells. Surprisingly IL-2 production terminated quickly in vivo and secretion was undetectable by 20-24 h in either cell type. We further demonstrated that this short duration of secretion can be influenced by cellular competition between Ag specific CD4(+) T cells. The consequences of competition were mimicked by reducing the strength of the antigenic stimulus. These data argue that early competition between T cells influences both the eventual frequency of IL-2 producers in the population and also the duration of their secretion, potentially by altering the strength or duration of the stimulus available to each T cell. PMID- 15128801 TI - Transcriptional activation of the SH2D2A gene is dependent on a cyclic adenosine 5'-monophosphate-responsive element in the proximal SH2D2A promoter. AB - The SH2D2A gene, encoding the T cell-specific adapter protein (TSAd), is rapidly induced in activated T cells. In this study we investigate the regulation of the SH2D2A gene in Jurkat T cells and in primary T cells. Reporter gene assays demonstrated that the proximal 1-kb SH2D2A promoter was constitutively active in Jurkat TAg T cells and, to a lesser extent, in K562 myeloid cells, Reh B cells, and 293T fibroblast cells. The minimal SH2D2A promoter was located between position -236 and -93 bp from the first coding ATG, and transcriptional activity in primary T cells depended on a cAMP response element (CRE) centered around position -117. Nuclear extracts from Jurkat TAg cells and activated primary T cells contained binding activity to this CRE, as observed in an EMSA. Consistent with this observation, we found that a cAMP analog was a very potent inducer of SH2D2A mRNA expression in primary T cells as measured by real-time RT-PCR. Furthermore, activation of SH2D2A expression by CD3 stimulation required cAMP dependent protein kinase activity. Thus, transcriptional regulation of the SH2D2A gene in activated T cells is critically dependent on a CRE in the proximal promoter region. PMID- 15128802 TI - Genetic makeup of the DR region in rhesus macaques: gene content, transcripts, and pseudogenes. AB - In the human population, five major HLA-DRB haplotypes have been identified, whereas the situation in rhesus macaques (Macaca mulatta) is radically different. At least 30 Mamu-DRB region configurations, displaying polymorphism with regard to number and combination of DRB loci present per haplotype, have been characterized. Until now, Mamu-DRB region genes have been studied mainly by genomic sequencing of polymorphic exon 2 segments. However, relatively little is known about the expression status of these genes. To understand which exon 2 segments may represent functional genes, full-length cDNA analyses of -DRA and DRB were initiated. In the course of the study, 11 cDRA alleles were identified, representing four distinct gene products. Amino acid replacements are confined to the leader peptide and cytoplasmatic tail, whereas residues of the alpha1 domain involved in peptide binding, are conserved between humans, chimpanzees, and rhesus macaques. Furthermore, from the 11 Mamu-DRB region configurations present in this panel, 28 cDRB alleles were isolated, constituting 12 distinct cDRA/cDRB configurations. Evidence is presented that a single configuration expresses maximally up to three -DRB genes. For some exon 2 DRB sequences, the corresponding transcripts could not be detected, rendering such alleles as probable pseudogenes. The full-length cDRA and cDRB sequences are necessary to construct Mhc class II tetramers, as well as transfectant cell lines. As the rhesus macaque is an important animal model in AIDS vaccine studies, the information provided in this communication is essential to define restriction elements and to monitor immune responses in SIV/simian human immunodeficiency virus-infected rhesus macaques. PMID- 15128803 TI - IL-4-induced GATA-3 expression is a time-restricted instruction switch for Th2 cell differentiation. AB - An initial activation signal via the TCR in a restricted cytokine environment is critical for the onset of Th cell development. Cytokines regulate the expression of key transcriptional factors, T-bet and GATA-3, which instruct the direction of Th1 and Th2 differentiation, through changes in chromatin conformation. In this study, we investigated the kinetics of IL-4-mediated signaling in a transgenic mouse, expressing human IL-4R on a mouse IL-4alphaR-deficient background. These experiments, allowing induction with human IL-4 at defined times, demonstrated that an IL-4 signal was required at the early stage of TCR-mediated T cell activation for lineage commitment to Th2, along with structural changes in chromatin, which take place in the conserved noncoding sequence-1 and -2 within the IL-4 locus. At later times, however, IL-4 failed to promote efficient Th2 differentiation and decondensation of chromatin, even though GATA-3 was clearly induced in the nuclei by IL-4 stimulation. Moreover, IL-4-mediated Th2 instruction was independent from cell division mediated by initial TCR stimulation. The role of IL-4 signaling may have a time restriction during Th2 differentiation. In late stages of initial T cell activation, the chromatin structure of the IL-4 locus retains condensation state. These results demonstrate that IL-4-induced GATA-3 expression is time-restriction switch for Th2 differentiation. PMID- 15128804 TI - Fusion of two malaria vaccine candidate antigens enhances product yield, immunogenicity, and antibody-mediated inhibition of parasite growth in vitro. AB - A Plasmodium falciparum chimeric protein 2.9 (PfCP-2.9) was constructed consisting of the C-terminal regions of two leading malaria vaccine candidates, domain III of apical membrane ag-1 (AMA-1) and 19-kDa C-terminal fragment of the merozoite surface protein 1 (MSP1). The PfCP-2.9 was produced by Pichia pastoris in secreted form with a yield of 2600 mg/L and approximately 1 g/L of final product was obtained from a three-step purification process. Analysis of conformational properties of the chimeric protein showed that all six conformational mAbs interacted with the recombinant protein were reduction sensitive, indicating that fusion of the two cysteine-rich proteins retains critical conformational epitopes. PfCP-2.9 was found to be highly immunogenic in rabbits as well as in rhesus monkeys (Macaca mulatta). The chimeric protein induced both anti-MSP1-19 and anti-AMA-1(III) Abs at levels 11- and 18-fold higher, respectively, than individual components did. Anti-PfCP-2.9 sera from both rabbits and rhesus monkeys almost completely inhibited in vitro growth of the P. falciparum FCC1/HN and 3D7 lines when tested at a 6.7-fold dilution. It was shown that the inhibition is dependent on the presence of Abs to the chimeric protein and their disulfide bond-dependent conformations. Moreover, the activity was mediated by a combination of growth-inhibitory Abs generated by the individual MSP1-19 and AMA-1(III) of PfCP-2.9. The combination of the extremely high yield of the protein and enhancement of its immune response provides a basis to develop an effective and affordable malaria vaccine. PMID- 15128805 TI - Structures of HLA-A*1101 complexed with immunodominant nonamer and decamer HIV-1 epitopes clearly reveal the presence of a middle, secondary anchor residue. AB - HLA-A*1101 is one of the most common human class I alleles worldwide. An increased frequency of HLA-A*1101 has been observed in cohorts of female sex workers from Northern Thailand who are highly exposed to HIV-1 and yet have remained persistently seronegative. In view of this apparent association of HLA A*1101 with resistance to acquisition of HIV-1 infection, and given the importance of eliciting strong CTL responses to control and eliminate HIV-1, we have determined the crystal structure of HLA-A*1101 complexed with two immunodominant HIV-1 CTL epitopes: the nonamer reverse transcriptase(313-321) (AIFQSSMTK) and decamer Nef(73-82) (QVPLRPMTYK) peptides. The structures confirm the presence of primary anchor residues P2-Ile/-Val and P9-/P10-Lys, and also clearly reveal the presence of secondary anchor residues P6-Ser for reverse transcriptase and P7-Met for Nef. The overall backbone conformation of both peptides is defined as two bulges that are separated by a more buried middle residue. In this study, we discuss how this topology may offer functional advantages in the selection and presentation of HIV-1 CTL epitopes by HLA-A*1101. Overall, this structural analysis permits a more accurate definition of the peptide-binding motif of HLA-A*1101, the characterization of its antigenic surface, and the correlation of molecular determinants with resistance to HIV-1 infection. These studies are relevant for the rational design of HLA-A*1101 restricted CTL epitopes with improved binding and immunological properties for the development of HIV-1 vaccines. PMID- 15128806 TI - Lipopolysaccharide-binding protein critically regulates lipopolysaccharide induced IFN-beta signaling pathway in human monocytes. AB - LPS binding to Toll-like receptor 4 induces a large number of genes through activation of NF-kappaB and IFN-regulatory factor-3 (IRF-3). However, no previous reports have tested the role of serum proteins in LPS-induced gene expression profiles. To investigate how serum proteins affect LPS-induced signaling, we investigated LPS-inducible genes in PBMC using an oligonucleotide probe-array system. Approximately 120 genes up-regulated by LPS were hierarchically divided into two clusters. Induction of one cluster, containing only IFN-inducible genes, was serum dependent. Real-time PCR analysis confirmed that IFN-inducible genes were induced only in the presence of serum, whereas inflammatory genes were induced both in the presence and absence of serum. Further analysis demonstrated that addition of LPS-binding protein (LBP), but not of soluble CD14 to the serum free medium enabled the induction of IFN-inducible genes and IFN-beta itself by LPS in human monocytes. The mRNAs for IFN-beta and IFN-inducible genes were induced by LPS only in the presence of serum from LBP(+/+) mice, and not in the presence of serum from LBP(-/-) mice. Blocking experiments also confirmed the involvement of LBP in this phenomenon. Immunoblotting analysis showed that phosphorylation of c-Jun N-terminal kinase, p38, IRF-3, tyrosine kinase 2, and STAT1 by LPS, but not of NF-kappaB and extracellular signal-regulated kinase was abrogated in the absence of LBP. This critical role for LBP implies the presence of possible mechanisms linking LBP to the intracellular signaling between Toll like receptor 4 and IRF-3, leading to the induction of IFN-beta by LPS. PMID- 15128807 TI - Lysine-dependent multipoint binding of the Borrelia burgdorferi virulence factor outer surface protein E to the C terminus of factor H. AB - Serum resistance, an important virulence determinant of Borrelia burgdorferi sensu lato strains belonging to the Borrelia afzelii and B. burgdorferi sensu stricto genotypes, is related to binding of the complement inhibitor factor H to the spirochete surface protein outer surface protein E (OspE) and its homologues. In this study, we show that the C-terminal short consensus repeats 18-20 of both human and mouse factor H bind to OspE. Analogously, factor H-related protein 1, a distinct plasma protein with three short consensus repeat domains homologous to those in factor H, bound to OspE. Deleting 15-aa residues (region V) from the C terminus of the OspE paralog P21 (a 20.7-kDa OspE-paralogous surface lipoprotein in the B. burgdorferi sensu stricto 297 strain) abolished factor H binding. However, C-terminal peptides from OspE, P21, or OspEF-related protein P alone and the C-terminal deletion mutants of P21 inhibited factor H binding to OspE only partially when compared with full-length P21 or its N-terminal mutant. Alanine substitution of amino acids in peptides from the key binding regions of the OspE family indicated that several lysine residues are required for factor H binding. Thus, the borrelial OspE family proteins bind the C inhibitor factor H via multiple sites in a lysine-dependent manner. The C-terminal site V (Ala(151) Lys(166)) is necessary, but not sufficient, for factor H binding in both rodents and humans. Identification of the necessary binding sites forms a basis for the development of vaccines that block the factor H-OspE interaction and thereby promote the killing of Borreliae. PMID- 15128808 TI - Toll-like receptor 4 dependence of innate and adaptive immunity to Salmonella: importance of the Kupffer cell network. AB - Mammalian cells recognize LPS from Gram-negative bacteria via the Toll-like receptor 4 (TLR4) complex. During experimental Salmonella infection, C3H/HeJ mice carrying a dominant-negative mutation in TLR4 exhibited delayed chemokine production, impaired NO generation, and attenuated cellular immune responses. However, dramatically enhanced bacterial growth within the Kupffer cell network before the recruitment of inflammatory cells appeared to be primarily responsible for the early demise of Salmonella-infected TLR4-deficient mice. LPS-TLR4 signaling plays an essential role in the generation of both innate and adaptive immune responses throughout the course of infection with Gram-negative bacteria. Alternative pattern-recognition receptors cannot completely compensate for the loss of TLR4, and compensation occurs at the expense of an increased microbial burden. PMID- 15128809 TI - Induction of HIV immunity in the genital tract after intranasal delivery of a MVA vector: enhanced immunogenicity after DNA prime-modified vaccinia virus Ankara boost immunization schedule. AB - Vaccines intended to prevent mucosal transmission of HIV should be able to induce multiple immune effectors in the host including Abs and cell-mediated immune responses at mucosal sites. The aim of this study was to characterize and to enhance the immunogenicity of a recombinant modified vaccinia virus Ankara (MVA) expressing HIV-1 Env IIIB Ag (MVAenv) inoculated in BALB/c mice by mucosal routes. Intravaginal inoculation of MVAenv was not immunogenic, whereas intranasally it induced a significant immune response to the HIV Ag. Intranasal codelivery of MVAenv plus cholera toxin (CT) significantly enhanced the cellular and humoral immune response against Env in the spleen and genitorectal draining lymph nodes, respectively. Heterologous DNAenv prime-MVAenv boost by intranasal immunization, together with CT, produced a cellular immune response in the spleen 10-fold superior to that in the absence of CT. A key finding of these studies was that both MVAenv/MVAenv and DNAenv/MVAenv schemes, plus CT, induced a specific mucosal CD8(+) T cell response in genital tissue and draining lymph nodes. In addition, both immunizations also generated systemic Abs, and more importantly, mucosal IgA and IgG Abs in vaginal washings. Specific secretion of beta chemokines was also generated by both immunizations, with a stronger response in mice immunized by the DNA-CT/MVA-CT regimen. Our findings are of relevance in the area of vaccine development and support the optimization of protocols of immunization based on MVA as vaccine vectors to induce mucosal immune responses against HIV. PMID- 15128810 TI - Immunological characteristics associated with the protective efficacy of antibodies to ricin. AB - A/B toxins, produced by bacteria and plants, are among the deadliest molecules known. The B chain binds the cell, whereas the A chain exerts the toxic effect. Both anti-A chain and anti-B chain Abs can neutralize toxins in vivo and in vitro. B chain Abs block binding of the toxin to the cell. It is not known how anti-A chain Abs function. Working with ricin toxin, we demonstrate that immunization with A chain induces greater protection than immunization with B chain. A panel of mAbs, binding to A chain, B chain, or both chains, has been produced and characterized. Immunologic characteristics evaluated include isotype, relative avidity, and epitope specificity. The ability to inhibit ricin enzymatic or cell binding activity was studied, as was the ability to block ricin mediated cellular cytotoxicity on human and murine cell lines. Finally, the in vivo protective efficacy of the Abs in mice was studied. The Ab providing the greatest in vivo protective efficacy was directed against the A chain. It had the greatest relative avidity and the greatest ability to block enzymatic function and neutralize cytotoxicity. Interestingly, we also obtained an anti-A chain Ab that bound with high avidity, blocked enzymatic activity, did not neutralize cytotoxicity, and actually enhanced the in vivo toxicity of ricin. Anti-A chain Abs with moderate avidity had no in vivo effect, nor did any anti-B chain Abs. PMID- 15128811 TI - Filaria-induced immune evasion: suppression by the infective stage of Brugia malayi at the earliest host-parasite interface. AB - To assess the physiologic interactions between the infective stage of Brugia malayi--one of the extracellular parasites responsible for lymphatic filariasis in humans--and the APC with which they come in contact during their development and routes of travel, we have investigated the interaction between the infective stage (L3) of B. malayi and human Langerhans cells (LC) in the skin. Our data indicate that live L3 result in increased migration of LC from the epidermis without affecting the viability of these cells and up-regulation of the IL-18 cytokine involved in LC migration. Live L3 also result in down-regulation of MHC class I and II on the LC cell surface. Additionally, microarray data indicate that live L3 significantly down-regulated expression of IL-8 as well as of multiple genes involved in Ag presentation, reducing the capacity of LC to induce CD4(+) T cells in allogeneic MLR, and thus resulting in a decreased ability of LC to promote CD4(+) T cell proliferation and production of IFN-gamma and IL-10. These data suggest that L3 exert a down-regulatory response in epidermal LC that leads to a diminished capacity of these cells to activate CD4(+) T cells. PMID- 15128812 TI - Visualizing the viral burden: phenotypic and functional alterations of T cells and APCs during persistent infection. AB - Persistent viral infections continue to present major public health problems. Failure to achieve virus control confronts the immune system with a chronic viral burden that may involve immune cells themselves and directly compromise the functionality of effector lymphocytes and APCs. In this study we use the lymphocytic choriomeningitis virus system for persistent viral infection of its natural murine host and use analytical techniques for direct ex vivo visualization of virus-infected immune cells. We report that virtually all cells of the immune system can be infected, but the distribution of the viral burden is differentially allocated to lymphocyte and APC subsets of defined phenotypes. Importantly, the profile of immune cell infection found in the blood is broadly representative for the pattern of cellular infection in most organs and is independent of the presence of Abs or complement. By direct comparison of virus infected and uninfected cell subsets, we demonstrate that lymphocytic choriomeningitis virus-infected T cells show preferential activation, skewed cytokine profiles, and increased apoptosis. In contrast, increased activation of APCs is generalized and independent of the presence of viral Ag. Our data indicate that specific patterns of immune cell infection are associated with distinct forms of immunostimulatory and immunosuppressive alterations that may provide insights into autoimmune processes associated with infectious disease and offer clues for therapeutic interventions aimed at restoration of complete immunity. PMID- 15128813 TI - Mycobacterium tuberculosis-induced CXCR4 and chemokine expression leads to preferential X4 HIV-1 replication in human macrophages. AB - Opportunistic infections such as pulmonary tuberculosis (TB) increase local HIV-1 replication and mutation. As AIDS progresses, alteration of the HIV-1 gp120 V3 sequence is associated with a shift in viral coreceptor use from CCR5 (CD195) to CXCR4 (CD184). To better understand the effect of HIV/TB coinfection, we screened transcripts from bronchoalveolar lavage cells with high density cDNA arrays and found that CXCR4 mRNA is increased in patients with TB. Surprisingly, CXCR4 was predominately expressed on alveolar macrophages (AM). Mycobacterium tuberculosis infection of macrophages in vitro increased CXCR4 surface expression, whereas amelioration of disease reduced CXCR4 expression in vivo. Bronchoalveolar lavage fluid from TB patients had elevated levels of CCL4 (macrophage inflammatory protein-1beta), CCL5 (RANTES), and CX3CL1 (fractalkine), but not CXCL12 (stromal derived factor-1alpha). We found that M. tuberculosis infection of macrophages in vitro increased viral entry and RT of CXCR4-using [corrected] HIV-1, but not of CCR5-using [corrected] HIV-1. Lastly, HIV-1 derived from the lung contains CD14, suggesting that they were produced in AM. Our results demonstrate that TB produces a permissive environment for replication of CXCR4-using virus by increasing CXCR4 expression in AM and for suppression of CCR5-using HIV-1 by increasing CC chemokine expression. These changes explain in part why TB accelerates the course of AIDS. CXCR4 inhibitors are a rational therapeutic approach in HIV/TB coinfection. PMID- 15128814 TI - IgA class switch occurs in the organized nasopharynx- and gut-associated lymphoid tissue, but not in the diffuse lamina propria of airways and gut. AB - Secretory IgA plays a crucial role in the host immune response as a first line of defense. A recent demonstration of in situ IgA class switching in intestinal lamina propria provided an opportunity to reconsider the model for the homing of IgA-committed B cells characterized by distinctive trafficking patterns to effector sites. Those effector sites depend on the organized mucosa-associated lymphoid tissues as their site of induction. In this report we show the preferential presence of IgM(+)B220(+) and IgA(+)B220(+) cells belonging to pre- and post-IgA isotype class-switched cells in the organized mucosa-associated lymphoid tissues, such as nasopharynx-associated lymphoid tissues, isolated lymphoid follicles, and Peyer's patches, and the defect of those populations in the diffuse effector tissues, such as the nasal passage and intestinal lamina propria. Consistent with these findings, the expressions of a series of IgA isotype class switch recombination-related molecules, including activation induced cytidine deaminase, Ialpha-C micro circle transcripts, and Ialpha-C micro circle transcripts, were selectively detected in these organized mucosa associated lymphoid structures, but not in the diffuse mucosal effector sites. Taken together, these findings suggest that IgA isotype class switching occurs only in the organized mucosa-associated lymphoid organs (e.g., nasopharynx associated lymphoid tissues, isolated lymphoid follicles, and Peyer's patches), but not in the diffuse effector tissues of the upper respiratory and gastrointestinal tracts. PMID- 15128815 TI - Cellular and humoral immunity against vaccinia virus infection of mice. AB - Despite the widespread use of vaccinia virus (VV) as a vector for other Ags and as the smallpox vaccine, there is little information available about the protective components of the immune response following VV infection. In this study, protection against wild-type VV was evaluated in mice with respect to the relative contributions of CD8(+) T cells vs that of CD4(+) T cells and Ab. C57BL/6 mice primed with the Western Reserve strain of VV mount significant IgM and IgG Ab responses, specific cytotoxic T cell responses, IFN-gamma responses in CD4(+) and CD8(+) T cells, and effectively clear the virus. This protection was abrogated by in vivo depletion of CD4(+) T cells or B cells in IgH(-/-) mice, but was not sensitive to CD8(+) T cell depletion alone. However, a role for CD8(+) T cells in primary protection was demonstrated in MHC class II(-/-) mice, where depleting CD8(+) T cells lead to increase severity of disease. Unlike control MHC class II(-/-) mice, the group depleted of CD8(+) T cells developed skin lesions on the tail and feet and had adrenal necrosis. Adoptive transfer experiments also show CD8(+) T cells can mediate protective memory. These results collectively show that both CD4(+) and CD8(+) T cell-mediated immunity can contribute to protection against VV infection. However, CD4(+) T cell-dependent anti-virus Ab production plays a more important role in clearing virus following acute infection, while in the absence of Ab, CD8(+) T cells can contribute to protection against disease. PMID- 15128816 TI - Mycobacterium tuberculosis inhibits macrophage responses to IFN-gamma through myeloid differentiation factor 88-dependent and -independent mechanisms. AB - Mycobacterium tuberculosis overcomes macrophage bactericidal activities and persists intracellularly. One mechanism by which M. tuberculosis avoids macrophage killing might be through inhibition of IFN-gamma-mediated signaling. In this study we provide evidence that at least two distinct components of M. tuberculosis, the 19-kDa lipoprotein and cell wall peptidoglycan (contained in the mycolylarabinogalactan peptidoglycan (mAGP) complex), inhibit macrophage responses to IFN-gamma at a transcriptional level. Moreover, these components engage distinct proximal signaling pathways to inhibit responses to IFN-gamma: the 19-kDa lipoprotein inhibits IFN-gamma signaling in a Toll-like receptor (TLR)2-dependent and myeloid differentiation factor 88-dependent fashion whereas mAGP inhibits independently of TLR2, TLR4, and myeloid differentiation factor 88. In addition to inhibiting the induction of specific IFN-gamma responsive genes, the 19-kDa lipoprotein and mAGP inhibit the ability of IFN-gamma to activate murine macrophages to kill virulent M. tuberculosis without inhibiting production of NO. These results imply that inhibition of macrophage responses to IFN-gamma may contribute to the inability of an apparently effective immune response to eradicate M. tuberculosis. PMID- 15128817 TI - Dendritic cells (DC) activated by CpG DNA ex vivo are potent inducers of host resistance to an intracellular pathogen that is independent of IL-12 derived from the immunizing DC. AB - We used the model of murine leishmaniasis to evaluate the signals enabling Ag pulsed dendritic cells (DC) to prime a protective Th1 response in vivo. Bone marrow-derived DC (BMDC) that had been activated by TNF-alpha or CD40 ligation were not able to induce protection against leishmaniasis in susceptible BALB/c mice. In contrast, all mice vaccinated with a single dose of Leishmania major Ag pulsed BMDC stimulated by prior in vitro exposure to CpG-containing oligodeoxynucleotides (ODN) were completely protected, had a dramatic reduction in parasite burden, and developed an Ag-specific Th1 response. Importantly, systemic administration of CpG ODN was not required. Protection mediated by ex vivo CpG ODN-activated and Ag-pulsed DC was solid, as documented by resistance to reinfection with a higher parasite dose, and long-lasting, as immunized mice were still protected against L. major challenge 16 wk after vaccination. A significantly increased level of protection could also be elicited in resistant C57BL/6 mice. Surprisingly, IL-12 expression by the immunizing BMDC was not required for induction of host resistance. In contrast, the availability of IL-12 derived from recipient cells was essential for the initial triggering of protective immunity by transferred BMDC. Together, these findings demonstrate that the type of stimulatory signal is critical for activating the potential of DC to induce a Th1 response in vivo that confers complete protection against an intracellular pathogen. Moreover, they show that the impact of activated DC on the initiation of a protective Th cell response in vivo may be independent of their ability to produce IL-12. PMID- 15128819 TI - Mouse strain susceptibility to trypanosome infection: an arginase-dependent effect. AB - We previously reported that macrophage arginase inhibits NO-dependent trypanosome killing in vitro and in vivo. BALB/c and C57BL/6 mice are known to be susceptible and resistant to trypanosome infection, respectively. Hence, we assessed the expression and the role of inducible NO synthase (iNOS) and arginase in these two mouse strains infected with Trypanosoma brucei brucei. Arginase I and arginase II mRNA expression was higher in macrophages from infected BALB/c compared with those from C57BL/6 mice, whereas iNOS mRNA was up-regulated at the same level in both phenotypes. Similarly, arginase activity was more important in macrophages from infected BALB/c vs infected C57BL/6 mice. Moreover, increase of arginase I and arginase II mRNA levels and of macrophage arginase activity was directly induced by trypanosomes, with a higher level in BALB/c compared with C57BL/6 mice. Neither iNOS expression nor NO production was stimulated by trypanosomes in vitro. The high level of arginase activity in T. brucei brucei-infected BALB/c macrophages strongly inhibited macrophage NO production, which in turn resulted in less trypanosome killing compared with C57BL/6 macrophages. NO generation and parasite killing were restored to the same level in BALB/c and C57BL/6 macrophages when arginase was specifically inhibited with N(omega)-hydroxy-nor-L arginine. In conclusion, host arginase represents a marker of resistance/susceptibility to trypanosome infections. PMID- 15128818 TI - Immunogenicity of recombinant adenovirus serotype 35 vaccine in the presence of pre-existing anti-Ad5 immunity. AB - The high prevalence of pre-existing immunity to adenovirus serotype 5 (Ad5) in human populations may substantially limit the immunogenicity and clinical utility of recombinant Ad5 vector-based vaccines for HIV-1 and other pathogens. A potential solution to this problem is to use vaccine vectors derived from adenovirus (Ad) serotypes that are rare in humans, such as Ad35. However, cross reactive immune responses between heterologous Ad serotypes have been described and could prove a major limitation of this strategy. In particular, the extent of immunologic cross-reactivity between Ad5 and Ad35 has not previously been determined. In this study we investigate the impact of pre-existing anti-Ad5 immunity on the immunogenicity of candidate rAd5 and rAd35 vaccines expressing SIV Gag in mice. Anti-Ad5 immunity at levels typically found in humans dramatically blunted the immunogenicity of rAd5-Gag. In contrast, even high levels of anti-Ad5 immunity did not substantially suppress Gag-specific cellular immune responses elicited by rAd35-Gag. Low levels of cross-reactive Ad5/Ad35 specific CD4(+) T lymphocyte responses were observed, but were insufficient to suppress vaccine immunogenicity. These data demonstrate the potential utility of Ad35 as a candidate vaccine vector that is minimally suppressed by anti-Ad5 immunity. Moreover, these studies suggest that using Ad vectors derived from immunologically distinct serotypes may be an effective and general strategy to overcome the suppressive effects of pre-existing anti-Ad immunity. PMID- 15128820 TI - Dendritic cells from mice neonatally vaccinated with modified vaccinia virus Ankara transfer resistance against herpes simplex virus type I to naive one-week old mice. AB - Modified vaccinia Ankara (MVA) is an attenuated virus. MVA induces the production of IFN and Flt3-L (FL), which results in the expansion of dendritic cells (DC) and enhanced resistance against viral infections. We report on the interplay among IFN, FL, and DC in the resistance against heterologous virus after injection of neonatal mice with MVA. The induction of serum FL was tested on day 2, and the expansion of DC was tested 1 wk after treatment with MVA. At this time point the resistance against infection with heterologous virus was also determined. After MVA treatment, serum FL was enhanced, and DC, including plasmacytoid cells in spleen, were increased in number. Mice that lacked functional IFN type I and II systems failed to increase both the concentration of FL and the number of DC. Treatment with MVA enhanced resistance against HSV-1 in wild-type animals 100-fold, but animals without a functional IFN system were not protected. Transfer of CD11c(+) cells from MVA-treated mice into naive animals protected against lethal infection with HSV-1. Thus, although the increased resistance could be largely attributed to the increase in activation of IFN producing plasmacytoid cells, this, in turn, depends on a complex interplay between the DC and T cell systems involving both FL and IFNs. PMID- 15128821 TI - Viral FLIP impairs survival of activated T cells and generation of CD8+ T cell memory. AB - Viral FLIPs (vFLIPs) interfere with apoptosis signaling by death-domain containing receptors in the TNFR superfamily (death receptors). In this study, we show that T cell-specific transgenic expression of MC159-vFLIP from the human Molluscum contagiosum virus blocks CD95-induced apoptosis in thymocytes and peripheral T cells, but also impairs postactivation survival of in vitro activated primary T cells despite normal early activation parameters. MC159 vFLIP impairs T cell development to a lesser extent than does Fas-associated death domain protein deficiency or another viral FLIP, E8. In the periphery, vFLIP expression leads to a specific deficit of functional memory CD8(+) T cells. After immunization with a protein Ag, Ag-specific CD8(+) T cells initially proliferate, but quickly disappear and fail to produce Ag-specific memory CD8(+) T cells. Viral FLIP transgenic mice exhibit impaired CD8(+) T cell responses to lymphocytic choriomeningitis virus and Trypanosoma cruzi infections, and a specific defect in CD8(+) T cell recall responses to influenza virus was seen. These results suggest that vFLIP expression in T cells blocks signals necessary for the sustained survival of CD8(+) T cells and the generation of CD8(+) T cell memory. Through this mechanism, vFLIP proteins expressed by T cell tropic viruses may impair the CD8(+) T cell immune responses directed against them. PMID- 15128822 TI - Dual role of TLR2 and myeloid differentiation factor 88 in a mouse model of invasive group B streptococcal disease. AB - Toll-like receptors (TLRs) are involved in pathogen recognition by the innate immune system. Different TLRs and the adaptor molecule myeloid differentiation factor 88 (MyD88) were previously shown to mediate in vitro cell activation induced by group B streptococcus (GBS). The present study examined the potential in vivo roles of TLR2 and MyD88 during infection with GBS. When pups were infected locally with a low bacterial dose, none of the TLR2- or MyD88-deficient mice, but all of the wild-type ones, were able to prevent systemic spread of GBS from the initial focus. Bacterial burden was higher in MyD88- than in TLR2 deficient mice, indicating a more profound defect of host defense in the former animals. In contrast, a high bacterial dose induced high level bacteremia in both mutant and wild-type mice. Under these conditions, however, TLR2 or MyD88 deficiency significantly protected mice from lethality, concomitantly with decreased circulating levels of TNF-alpha and IL-6. Administration of anti-TNF alpha Abs to wild-type mice could mimic the effects of TLR2 or MyD88 deficiency and was detrimental in the low dose model, but protective in the high dose model. In conclusion, these data highlight a dual role of TLR2 and MyD88 in the host defense against GBS sepsis and strongly suggest TNF-alpha as the molecular mediator of bacterial clearance and septic shock. PMID- 15128823 TI - Augmentation of chemotherapy-induced cytokine production by expression of the platelet-activating factor receptor in a human epithelial carcinoma cell line. AB - In addition to their known cytotoxic effects, chemotherapeutic agents can trigger cytokine production in tumor cells. Moreover, many chemotherapeutic agents are potent pro-oxidative stressors. Although the lipid mediator platelet-activating factor (PAF) is synthesized in response to oxidative stress, and many epidermal carcinomas express PAF receptors (PAF-R) linked to cytokine production, it is not known whether PAF is involved in chemotherapeutic agent-induced cytokine production. These studies examined the role of the PAF system in chemotherapy mediated cytokine production using a model system created by retroviral-mediated transduction of the PAF-R-negative human epidermal carcinoma cell line KB with the human PAF-R. The presence of the PAF-R in KB cells resulted in augmentation of the production of cytokines IL-8 and TNF-alpha induced by the chemotherapeutic agents etoposide and mitomycin C. These effects were specific for the PAF-R, as expression of the G protein-coupled receptor for fMLP did not affect chemotherapeutic agent-induced cytokine production. Moreover, ablation of the native PAF-R in the epithelial cell line HaCaT using an inducible antisense PAF-R strategy inhibited etoposide-induced cytokine production. Oxidative stress and the transcription factor NF-kappaB were found to be involved in this augmentative effect, because it was mimicked by the oxidant tert-butyl-hydroperoxide, which was blocked both by antioxidants and by inhibition of the NFkappaB pathway using a super-repressor IkappaBM mutant. These studies provide evidence for a novel pathway by which the epidermal PAF-R can augment chemotherapy-induced cytokine production through an NF-kappaB-dependent process. PMID- 15128824 TI - Transient and selective NF-kappa B p65 serine 536 phosphorylation induced by T cell costimulation is mediated by I kappa B kinase beta and controls the kinetics of p65 nuclear import. AB - Full transcriptional activity of the nuclear, DNA-bound form of NF-kappaB requires additional posttranslational modifications. In this study, we systematically mapped the T cell costimulation-induced phosphorylation sites within the C-terminal half of the strongly trans-activating NF-kappaB p65 subunit and identified serine 536 as the main phosphorylation site. The transient kinetics of serine 536 phosphorylation paralleled the kinetics of IkappaBalpha and IkappaB kinase (IKK) phosphorylation and also mirrored the principle of T cell costimulation. The TCR-induced pathway leading to serine 536 phosphorylation is regulated by the kinases Cot (Tpl2), receptor interacting protein, protein kinase Ctheta, and NF-kappaB-inducing kinase, but is independent from the phosphatidylinositol 3-kinase/Akt signaling pathway. Loss-of-function and gain-of function experiments showed phosphorylation of p65 serine 536 by IKKbeta, but not by IKKalpha. Phosphorylation occurs within the cytoplasmic and intact NF kappaB/IkappaBalpha complex and requires prior phosphorylation of IkappaBalpha at serines 32 and 36. Reconstitution of p65(-/-) cells either with wild-type p65 or a p65 mutant containing a serine to alanine mutation revealed the importance of this phosphorylation site for cytosolic IkappaBalpha localization and the kinetics of p65 nuclear import. PMID- 15128825 TI - Intracellular bacterial infection-induced IFN-gamma is critically but not solely dependent on Toll-like receptor 4-myeloid differentiation factor 88-IFN-alpha beta-STAT1 signaling. AB - Infection of murine bone marrow-derived macrophages (BMMphi) with Chlamydia pneumoniae induces IFN-alphabeta-dependent IFN-gamma secretion that leads to control of the intracellular bacterial growth. Enhanced growth of C. pneumoniae in Toll-like receptor (TLR) 4(-/-) and myeloid differentiation factor (MyD) 88(-/ ) (but not TLR2(-/-), TLR6(-/-), or TLR9(-/-)) BMMphi is shown in this study. Reduced accumulation of IFN-alpha and IFN-gamma mRNA was also observed in TLR4(-/ )- and MyD88(-/-)-infected cells. IL-1R and IL-18R signaling did not account for differences between MyD88(-/-) and wild-type BMMphi. Surprisingly, infection induced NF-kappaB activation as well as TNF-alpha, IL-1, or IL-6 mRNA expression were all normal in TLR4(-/-) and MyD88(-/-) cells. Phosphorylation of the transcription factor STAT1 during bacterial infection is IFN-alphabeta dependent, and necessary for increased IFN-gamma mRNA accumulation and chlamydial growth control. Signaling through common cytokine receptor gamma-chain and RNA-dependent protein kinase both mediated IFN-alphabeta-dependent enhancement of IFN-gamma mRNA levels. Accumulation of IFN-gamma mRNA and control of C. pneumoniae growth required NF-kappaB activation. Such NF-kappaB activation was independent of IFN alphabeta, STAT1, and RNA-dependent protein kinase. In summary, C. pneumoniae induced IFN-gamma expression in BMMphi is controlled by a TLR4-MyD88-IFN alphabeta-STAT1-dependent pathway, as well as by a TLR4-independent pathway leading to NF-kappaB activation. PMID- 15128826 TI - Studies of congenic lines in the Brown Norway rat model of Th2-mediated immunopathological disorders show that the aurothiopropanol sulfonate-induced immunological disorder (Aiid3) locus on chromosome 9 plays a major role compared to Aiid2 on chromosome 10. AB - Brown Norway (BN) rats treated with aurothiopropanol-sulfonate (Atps) constitute a model of Th2-mediated immunological disorders associated with elevated IgE responses and renal IgG deposits. Using F(2) offspring between Atps-susceptible BN and Atps-resistant Lewis rats, we had previously mapped three quantitative trait loci on chromosomes 9, 10, and 20 for which BN alleles increased susceptibility to Atps-induced immunological disorders (Aiid). In this study we have used congenic lines for the latter two quantitative trait loci, formerly called Atps2 and Atps3 and now named Aiid2 (chromosome 10) and Aiid3 (chromosome 9), for fine mapping and characterization of their impact on Atps-triggered reactions. In Aiid2 congenic lines, the gene(s) controlling part of the IgE response to Atps was mapped to an approximately 7-cM region, which includes the IL-4 cytokine gene cluster. Two congenic lines in which the introgressed segments shared only a portion of this 7-cM region, showed an intermediate IgE response, indicating the involvement of several genes within this region. Results from BN rats congenic for the Lewis Aiid3 locus, which we mapped to a 1.2-cM interval, showed a stronger effect of this region. In this congenic line, the Atps triggered IgE response was 10-fold lower than in the BN parental strain, and glomerular IgG deposits were either absent or dramatically reduced. Further genetic and functional dissections of these loci should provide insights into pathways that lead to Th2-adverse reactions. PMID- 15128827 TI - The transmembrane CXC-chemokine ligand 16 is induced by IFN-gamma and TNF-alpha and shed by the activity of the disintegrin-like metalloproteinase ADAM10. AB - The novel CXC-chemokine ligand 16 (CXCL16) functions as transmembrane adhesion molecule on the surface of APCs and as a soluble chemoattractant for activated T cells. In this study, we elucidate the mechanism responsible for the conversion of the transmembrane molecule into a soluble chemokine and provide evidence for the expression and shedding of CXCL16 by fibroblasts and vascular cells. By transfection of human and murine CXCL16 in different cell lines, we show that soluble CXCL16 is constitutively generated by proteolytic cleavage of transmembrane CXCL16 resulting in reduced surface expression of the transmembrane molecule. Inhibition experiments with selective hydroxamate inhibitors against the disintegrin-like metalloproteinases a disintegrin and metalloproteinase domain (ADAM)10 and ADAM17 suggest that ADAM10, but not ADAM17, is involved in constitutive CXCL16 cleavage. In addition, the constitutive cleavage of transfected human CXCL16 was markedly reduced in embryonic fibroblasts generated from ADAM10-deficient mice. By induction of murine CXCL16 in ADAM10-deficient fibroblasts with IFN-gamma and TNF-alpha, we show that endogenous ADAM10 is indeed involved in the release of endogenous CXCL16. Finally, the shedding of endogenous CXCL16 could be reconstituted by retransfection of ADAM10-deficient cells with ADAM10. Analyzing the expression and release of CXCXL16 by cultured vascular cells, we found that IFN-gamma and TNF-alpha synergize to induce CXCL16 mRNA. The constitutive shedding of CXCL16 from the endothelial cell surface is blocked by inhibitors of ADAM10 and is independent of additional inhibition of ADAM17. Hence, during inflammation in the vasculature, ADAM10 may act as a CXCL16 sheddase and thereby finely control the expression and function of CXCL16 in the inflamed tissue. PMID- 15128828 TI - 6-phosphogluconate dehydrogenase and glucose-6-phosphate dehydrogenase form a supramolecular complex in human neutrophils that undergoes retrograde trafficking during pregnancy. AB - Neutrophils from pregnant women display reduced neutrophil-mediated effector functions, such as reactive oxygen metabolite (ROM) release. Because the NADPH oxidase and NO synthase produce ROMs and NO, the availability of their substrate NADPH is a potential regulatory factor. NADPH is produced by glucose-6-phosphate dehydrogenase (G-6-PDase) and 6-phosphogluconate dehydrogenase (6-PGDase), which are the first two steps of the hexose monophosphate shunt (HMS). Using immunofluorescence microscopy, we show that 6-PGDase, like G-6-PDase, undergoes retrograde transport to the microtubule-organizing centers in neutrophils from pregnant women. In contrast, 6-PGDase is found in an anterograde distribution in cells from nonpregnant women. However, lactate dehydrogenase distribution is unaffected by pregnancy. Cytochemical studies demonstrated that the distribution of 6-PGDase enzymatic activity is coincident with 6-PGDase Ag. The accumulation of 6-PGDase at the microtubule-organizing centers could be blocked by colchicine, suggesting that microtubules are important in this enzyme's intracellular distribution. In situ kinetic studies reveal that the rates of 6-gluconate turnover are indistinguishable in samples from nonpregnant and pregnant women, suggesting that the enzyme is functionally intact. Resonance energy transfer experiments showed that 6-PGDase and G-6-PDase are in close physical proximity within cells, suggesting the presence of supramolecular enzyme complexes. We suggest that the retrograde trafficking of HMS enzyme complexes during pregnancy influences the dynamics of NADPH production by separating HMS enzymes from glucose-6-phosphate generation at the plasma membrane and, in parallel, reducing ROM and NO production in comparison with fully activated neutrophils from nonpregnant women. PMID- 15128829 TI - Inactivation of C5a anaphylatoxin by a peptide that is complementary to a region of C5a. AB - PL37 (RAARISLGPRCIKAFTE) is an antisense homology box peptide composed of aa 37 53 of C5a-anaphylatoxin and is considered to be the region essential for C5a function. Using a computer program, we designed the complementary peptides ASGAPAPGPAGPLRPMF (Pep-A) and ASTAPARAGLPRLPKFF (Pep-B). Pep-A bound to PL37 and to C5a with very slow dissociation as determined by analysis using surface plasmon resonance, whereas Pep-B failed to bind at all. C5a was inactivated by concentrations of 7 nM or more of Pep-A, and this concentration of Pep-A inhibited induction of intracellular Ca(2+) influx in neutrophils. Patch clamp electrophysiology experiments also showed the effectiveness of Pep-A in C5aR expressing neuroblastoma cells. Furthermore, Pep-A administration prevented rats from C5a-mediated rapid lethal shock induced by an Ab to a membrane inhibitor of complement after LPS sensitization. PMID- 15128830 TI - Ectopic CD40 ligand expression on B cells triggers intestinal inflammation. AB - Several studies indicate that CD4(+) T cells, macrophages, and dendritic cells initially mediate intestinal inflammation in murine models of human inflammatory bowel disease. However, the initial role of B cells in the development of intestinal inflammation remains unclear. In this study we present evidence that B cells can trigger intestinal inflammation using transgenic (Tg) mice expressing CD40 ligand (CD40L) ectopically on B cells (CD40L/B Tg). We demonstrated that CD40L/B Tg mice spontaneously developed severe transmural intestinal inflammation in both colon and ileum at 8-15 wk of age. In contrast, CD40L/B TgxCD40(-/-) double-mutant mice did not develop colitis, indicating the direct involvement of CD40-CD40L interaction in the development of intestinal inflammation. The inflammatory infiltrates consisted predominantly of massive aggregated, IgM positive B cells. These mice were also characterized by the presence of anti colon autoantibodies and elevated IFN-gamma production. Furthermore, although mice transferred with CD4(+) T cells alone or with both CD4(+) T and B220(+) B cells, but not B220(+) cells alone, from diseased CD40L/B Tg mice, develop colitis, mice transferred with B220(+) B cells from diseased CD40L/B Tg mice and CD4(+) T cells from wild-type mice also develop colitis, indicating that the Tg B cells should be a trigger for this colitis model, whereas T cells are involved as effectors. As it has been demonstrated that CD40L is ectopically expressed on B cells in some autoimmune diseases, the present study suggests the possible contribution of B cells in triggering intestinal inflammation in human inflammatory bowel disease. PMID- 15128831 TI - Mast cells, Fc epsilon RI, and IL-13 are required for development of airway hyperresponsiveness after aerosolized allergen exposure in the absence of adjuvant. AB - In certain models of allergic airway disease, mast cells facilitate the development of inflammation and airway hyper-responsiveness (AHR). To define the role of the high affinity IgE receptor (FcepsilonRI) in the development of AHR, mice with a disruption of the alpha subunit of the high affinity IgE receptor (FcepsilonRI(-/-)) were exposed on 10 consecutive days to nebulized OVA. Forty eight hours after the last nebulization, airway responsiveness was monitored by the contractile response of tracheal smooth muscle to electrical field stimulation (EFS). After the 10-day OVA challenge protocol, wild-type mice demonstrated increased responsiveness to EFS, whereas similarly challenged FcepsilonRI(-/-) mice showed a low response to EFS, similar to nonexposed animals. Further, allergen-challenged FcepsilonRI(-/-) mice showed less airway inflammation, goblet cell hyperplasia, and lower levels of IL-13 in lung homogenates compared with the controls. IL-13-deficient mice failed to develop an increased response to EFS or goblet cell hyperplasia after the 10-day OVA challenge. We transferred bone marrow-derived mast cells from wild-type mice to FcepsilonRI(-/-) mice 1 day before initiating the challenge protocol. After the 10-day OVA challenge, recipient FcepsilonRI(-/-) mice demonstrated EFS-induced responses similar to those of challenged wild-type mice. Transferred mast cells could be detected in tracheal preparations. These results show that FcepsilonRI is important for the development of AHR after an aerosolized allergen sensitization protocol and that this effect is mediated through FcepsilonRI on mast cells and production of IL-13 in the lung. PMID- 15128832 TI - The functional profile of primary human antiviral CD8+ T cell effector activity is dictated by cognate peptide concentration. AB - Antiviral CD8(+) T cells can elaborate at least two effector functions, cytokine production and cytotoxicity. Which effector function is elaborated can determine whether the CD8(+) T cell response is primarily inflammatory (cytokine producing) or antiviral (cytotoxic). In this study we demonstrate that cytotoxicity can be triggered at peptide concentrations 10- to 100-fold less than those required for cytokine production in primary HIV- and CMV-specific human CD8(+) T cells. Cytolytic granule exocytosis occurs at peptide concentrations insufficient to cause substantial TCR down-regulation, providing a mechanism by which a CD8(+) T cell could engage and lyse multiple target cells. TCR sequence analysis of virus specific cells shows that individual T cell clones can degranulate or degranulate and produce cytokine depending on the Ag concentration, indicating that response heterogeneity exists within individual CD8(+) T cell clonotypes. Thus, antiviral CD8(+) T cell effector function is determined primarily by Ag concentration and is not an inherent characteristic of a virus-specific CD8(+) T cell clonotype or the virus to which the response is generated. The inherent ability of viruses to induce high or low Ag states may be the primary determinant of the cytokine vs cytolytic nature of the virus-specific CD8(+) T cell response. PMID- 15128833 TI - Attenuation of experimental autoimmune encephalomyelitis and nonimmune demyelination by IFN-beta plus vitamin B12: treatment to modify notch-1/sonic hedgehog balance. AB - Interferon-beta is a mainstay therapy of demyelinating diseases, but its effects are incomplete in human multiple sclerosis and several of its animal models. In this study, we demonstrate dramatic improvements of clinical, histological, and laboratory parameters in in vivo mouse models of demyelinating disease through combination therapy with IFN-beta plus vitamin B(12) cyanocobalamin (B(12)CN) in nonautoimmune primary demyelinating ND4 (DM20) transgenics, and in acute and chronic experimental autoimmune encephalomyelitis in SJL mice. Clinical improvement (p values <0.0001) was paralleled by near normal motor function, reduced astrocytosis, and reduced demyelination. IFN-beta plus B(12)CN enhanced in vivo and in vitro oligodendrocyte maturation. In vivo and in vitro altered expression patterns of reduced Notch-1 and enhanced expression of sonic hedgehog and its receptor were consistent with oligodendrocyte maturation and remyelination. IFN-beta-B(12)CN combination therapy may be promising for the treatment of multiple sclerosis. PMID- 15128834 TI - Generation and regulation of human Th1-biased immune responses in vivo: a critical role for IL-4 and IL-10. AB - Tissue damage in many human autoimmune diseases is mediated by activated autoantigen-specific Th1 cells. Delineation of the regulatory mechanisms controlling a Th1-biased human immune reaction and its pathologic potential is, therefore, a critical step in the understanding of autoimmune diseases. In this study, we introduce a novel means to investigate human Th1-biased immune responses in vivo. Intraperitoneal injection of human mononuclear cells into immunodeficient mice generates a xenogeneic Th1-biased human immune response characterized by systemic inflammation and leukocytic infiltrates with a granuloma-like architecture in the liver, and the perigastrointestinal and perirenal fatty tissue. Th1 cell activation was dependent on the presence of APCs and could be blocked by cyclosporine. Importantly, neutralization of endogenously produced IL-4 and IL-10 markedly exaggerated the immune response, whereas exogenous IL-4 and IL-10 inhibited systemic Th1 immunity. Thus, the model described in this paper presents a useful means to analyze the regulation of human immune reactions in an in vivo situation. The results suggest that both IL 4 and IL-10 contribute to controlling the development of a human Th1-biased immune reaction. PMID- 15128835 TI - CD4+CD25bright regulatory T cells actively regulate inflammation in the joints of patients with the remitting form of juvenile idiopathic arthritis. AB - This study investigates the role of CD4(+)CD25(+) regulatory T cells during the clinical course of juvenile idiopathic arthritis (JIA). Persistent oligoarticular JIA (pers-OA JIA) is a subtype of JIA with a relatively benign, self-remitting course while extended oligoarticular JIA (ext-OA JIA) is a subtype with a much less favorable prognosis. Our data show that patients with pers-OA JIA display a significantly higher frequency of CD4(+)CD25(bright) T cells with concomitant higher levels of mRNA FoxP3 in the peripheral blood than ext-OA JIA patients. Furthermore, while numbers of synovial fluid (SF) CD4(+)CD25(bright) T cells were equal in both patient groups, pers-OA JIA patients displayed a higher frequency of CD4(+)CD25(int) T cells and therefore of CD4(+)CD25(total) in the SF than ext OA JIA patients. Analysis of FoxP3 mRNA levels revealed a high expression in SF CD4(+)CD25(bright) T cells of both patient groups and also significant expression of FoxP3 mRNA in the CD4(+)CD25(int) T cell population. The CD4(+)CD25(bright) cells of both patient groups and the CD4(+)CD25(int) cells of pers-OA JIA patients were able to suppress responses of CD25(neg) cells in vitro. A markedly higher expression of CTLA-4, glucocorticoid-induced TNFR, and HLA-DR on SF CD4(+)CD25(bright) T regulatory (Treg) cells compared with their peripheral counterparts suggests that the CD4(+)CD25(+) Treg cells may undergo maturation in the joint. In correlation with this mature phenotype, the SF CD4(+)CD25(bright) T cells showed an increased regulatory capacity in vitro compared with peripheral blood CD4(+)CD25(bright) T cells. These data suggest that CD4(+)CD25(bright) Treg cells play a role in determining the patient's fate toward either a favorable or unfavorable clinical course of disease. PMID- 15128836 TI - Xenobiotic-induced loss of tolerance in rabbits to the mitochondrial autoantigen of primary biliary cirrhosis is reversible. AB - Previous work has demonstrated that immunization of rabbits with the xenobiotic 6 bromohexanoate coupled to BSA breaks tolerance and induces autoantibodies to mitochondria in rabbits. Such immunized rabbits develop high-titer Abs to pyruvate dehydrogenase complex (PDC)-E2, the major autoantigen of primary biliary cirrhosis. In efforts to map the fine specificity of these autoantibodies, rabbits were immunized biweekly with 6-bromohexanoate-BSA and screened for reactivity using a unique xenobiotic-peptide-agarose microarray platform with an emphasis on identifying potential structures that mimic the molecular image formed by the association of lipoic acid with the immunodominant PDC-E2 peptide. Essentially, a total of 23 xenobiotics and lipoic acid were coupled to the 12-mer peptide backbones, PDC, a mutant PDC, and albumin. As expected, we succeeded in breaking tolerance using this small organic molecule coupled to BSA. However, unlike multiple experimental methods of breaking tolerance, we report in this study that, following continued immunization, the rabbits recover tolerance. With repeated immunization, the response to the rPDC-E2 protein increased with a gradual reduction in autoantibodies against the lipoic acid-peptide, i.e., the primary tolerance-breaking autoantigen. Detailed analysis of this system may provide strategies on how to restore tolerance in patients with autoimmune disease. PMID- 15128837 TI - Antiviral cytokines induce hepatic expression of the granzyme B inhibitors, proteinase inhibitor 9 and serine proteinase inhibitor 6. AB - Expression of the granzyme B inhibitors, human proteinase inhibitor 9 (PI-9), or the murine orthologue, serine proteinase inhibitor 6 (SPI-6), confers resistance to CTL or NK killing by perforin- and granzyme-dependent effector mechanisms. In light of prior studies indicating that virally infected hepatocytes are selectively resistant to this CTL effector mechanism, the present studies investigated PI-9 and SPI-6 expression in hepatocytes and hepatoma cells in response to adenoviral infection and to cytokines produced during antiviral immune responses. Neither PI-9 nor SPI-6 expression was detected by immunoblotting in uninfected murine or human hepatocytes. Similarly, human Huh-7 hepatoma cells were found to express only very low levels of PI-9 relative to levels detected in perforin- and granzyme-resistant CTL or lymphokine-activated killer cells. Following in vivo adenoviral infection or in vitro culture with IFN alphabeta or IFN-gamma, SPI-6 expression was induced in murine hepatocytes. Similarly, after culture with IFN-alpha, induction of PI-9 mRNA and protein expression was observed in human hepatocytes and Huh-7 cells. IFN-gamma and TNF alpha also induced 4- to 10-fold higher levels of PI-9 mRNA expression in Huh-7 cells, whereas levels of mRNA encoding a related serine proteinase inhibitor, proteinase inhibitor 8, were unaffected by culture of Huh-7 cells with IFN-alpha, IFN-gamma, or TNF-alpha. These findings indicate that cytokines that promote antiviral cytopathic responses also regulate expression of the cytoprotective molecules, PI-9 and SPI-6, in hepatocytes that are potential targets of CTL and NK effector mechanisms. PMID- 15128838 TI - Lack of galactose-alpha-1,3-galactose expression on porcine endothelial cells prevents complement-induced lysis but not direct xenogeneic NK cytotoxicity. AB - The galactose-alpha-1,3-galactose (alphaGal) carbohydrate epitope is expressed on porcine, but not human cells, and therefore represents a major target for preformed human anti-pig natural Abs (NAb). Based on results from pig-to-primate animal models, NAb binding to porcine endothelial cells will likely induce complement activation, lysis, and hyperacute rejection in pig-to-human xenotransplantation. Human NK cells may also contribute to innate immune responses against xenografts, either by direct recognition of activating molecules on target cells or by FcgammaRIII-mediated xenogeneic Ab-dependent cellular cytotoxicity (ADCC). The present study addressed the question as to whether the lack of alphaGal protects porcine endothelial cells from NAb/complement-induced lysis, direct xenogeneic NK lysis, NAb-dependent ADCC, and adhesion of human NK cells under shear stress. Homologous recombination, panning, and limiting dilution cloning were used to generate an alphaGal-negative porcine endothelial cell line, PED2*3.51. NAb/complement-induced xenogeneic lysis of PED2*3.51 was reduced by an average of 86% compared with the alphaGal-positive phenotype. PED2*3.51 resisted NK cell-mediated ADCC with a reduction of lysis ranging from 30 to 70%. However, direct xenogeneic lysis of PED2*3.51, mediated either by freshly isolated or IL-2-activated human NK cells or the NK cell line NK92, was not reduced. Furthermore, adhesion of IL-2-activated human NK cells did not rely on alphaGal expression. In conclusion, removal of alphaGal leads to a clear reduction in complement-induced lysis and ADCC, but does not resolve adhesion of NK cells and direct anti-porcine NK cytotoxicity, indicating that alphaGal is not a dominant target for direct human NK cytotoxicity against porcine cells. PMID- 15128839 TI - Type I regulatory T cells specific for desmoglein 3 are more frequently detected in healthy individuals than in patients with pemphigus vulgaris. AB - Pemphigus vulgaris (PV) is a severe autoimmune bullous skin disorder and is primarily associated with circulating autoantibodies against desmoglein 3 (Dsg3) that are presumably regulated by Th cells. The aim of this study was to identify Dsg3-specific T regulatory (Tr) cells that may help to maintain and restore natural tolerance against Dsg3. Dsg3-responsive IL-10-secreting Tr1 cells were isolated by MACS cytokine secretion assay from healthy carriers of the PV associated HLA class II alleles, DRB1*0402 and DQB1*0503, but were only rarely detected in PV patients. The Dsg3-specific Tr1 cells secreted IL-10, TGF-beta, and IL-5 upon Ag stimulation, proliferated in response to IL-2 but not to Dsg3 or mitogenic stimuli, and inhibited the proliferative response of Dsg3- and tetanus toxoid-responsive Th clones in an Ag-specific (Dsg3) and cell number-dependent manner. Moreover, their inhibitory effect was blocked by Ab against IL-10, TGF beta, and by paraformaldehyde fixation. These observations strongly suggest that 1) Dsg3-responsive Tr1 cells predominate in healthy individuals, 2) their growth requires the presence of IL-2, and 3) they exert their Dsg3-dependent inhibitory function by the secretion of IL-10 and TGF-beta. Because autoaggressive T cells responsive to identical epitopes of Dsg3 were recently found both in PV patients and healthy individuals, the identified Tr1 cells may be critically involved in the maintenance and restoration of tolerance against Dsg3. PMID- 15128840 TI - IFN-alpha priming results in a gain of proinflammatory function by IL-10: implications for systemic lupus erythematosus pathogenesis. AB - Interleukin-10 is a predominantly anti-inflammatory cytokine that inhibits macrophage and dendritic cell function, but can acquire proinflammatory activity during immune responses. We investigated whether type I IFNs, which are elevated during infections and in autoimmune diseases, modulate the activity of IL-10. Priming of primary human macrophages with low concentrations of IFN-alpha diminished the ability of IL-10 to suppress TNF-alpha production. IFN-alpha conferred a proinflammatory gain of function on IL-10, leading to IL-10 activation of expression of IFN-gamma-inducible, STAT1-dependent genes such as IFN regulatory factor 1, IFN-gamma-inducible protein-10 (CXCL10), and monokine induced by IFN-gamma (CXCL9). IFN-alpha priming resulted in greatly enhanced STAT1 activation in response to IL-10, and STAT1 was required for IL-10 activation of IFN-gamma-inducible protein-10 and monokine induced by IFN-gamma expression in IFN-alpha-primed cells. In control, unprimed cells, IL-10 activation of STAT1 was suppressed by constitutive activity of protein kinase C and Src homology 2 domain-containing phosphatase 1. These results demonstrate that type I IFNs regulate the balance between IL-10 anti- and proinflammatory activity, and provide insight into molecular mechanisms that regulate IL-10 function. Gain of IL-10 proinflammatory functions may contribute to its pathogenic role in autoimmune diseases characterized by elevated type I IFN levels, such as systemic lupus erythematosus. PMID- 15128841 TI - The novel cyclophilin-binding drug sanglifehrin A specifically affects antigen uptake receptor expression and endocytic capacity of human dendritic cells. AB - Sanglifehrin A (SFA) is a recently developed immunosuppressant that belongs to the family of immunophilin-binding ligands. SFA is a cyclophilin A-binding immunosuppressive drug with a novel, but unidentified, mechanism of action. Several reports exist about the effect of SFA on T cells, but its effect on the initiators of the immune response, i.e., dendritic cells (DCs), is relatively unknown. Therefore, we examined the effect of SFA on the differentiation and function of human monocyte-derived DCs. Unlike the well-known cyclophilin A binding immunosuppressant cyclosporin A, which did not affect DC phenotype, differentiation of DCs in the presence of SFA resulted in CD14-CD1a DCs with normal DC morphology, viability, and a proper capacity to activate allogeneic T cells. However, DCs generated in the presence of SFA demonstrated reduced macropinocytosis and lectin-mediated endocytosis, which was in line with a decreased expression of C-type lectins, including mannose receptor, C1qRP, DC ASGPR, and especially, DC-SIGN. In contrast, FcalphaRI (CD89) and FcgammaRII (CD32) were increased by SFA. The explicit effect of SFA on the expression of Ag uptake receptors and Ag capture by DCs makes SFA unique among immunophilin binding immunosuppressive drugs. PMID- 15128842 TI - Molecular characterization of polygalacturonases as grass pollen-specific marker allergens: expulsion from pollen via submicronic respirable particles. AB - Grass pollen belong to the most important allergen sources involved in the elicitation of allergic asthma. We have isolated cDNAs coding for Bermuda grass (Cynodon dactylon) and timothy grass (Phleum pratense) pollen allergens, belonging to a family of pectin-degrading enzymes (i.e., polygalacturonases). The corresponding allergens, termed Cyn d 13 and Phl p 13, represent glycoproteins of approximately 42 kDa and isoelectric points of 7.5. rPhl p 13 was expressed in Escherichia coli and purified to homogeneity. Immunogold electron microscopy using rabbit anti-rPhl p 13 Abs demonstrated that in dry pollen group 13, allergens represent primarily intracellular proteins, whereas exposure of pollen to rainwater caused a massive release of cytoplasmic material containing submicronic particles of respirable size, which were coated with group 13 allergens. The latter may explain respiratory sensitization to group 13 allergens and represents a possible pathomechanism in the induction of asthma attacks after heavy rainfalls. rPhl p 13 was recognized by 36% of grass pollen allergic patients, showed IgE binding capacity comparable to natural Phl p 13, and induced specific and dose-dependent basophil histamine release. Epitope mapping studies localized major IgE epitopes to the C terminus of the molecule outside the highly conserved functional polygalacturonase domains. The latter result explains why rPhl p 13 contains grass pollen-specific IgE epitopes and may be used to diagnose genuine sensitization to grass pollen. Our finding that rabbit anti-rPhl p 13 Abs blocked patients' IgE binding to the allergen suggests that rPhl p 13 may be used for immunotherapy of sensitized patients. PMID- 15128843 TI - Proteinase-activated receptor 2-mediated potentiation of transient receptor potential vanilloid subfamily 1 activity reveals a mechanism for proteinase induced inflammatory pain. AB - Proteinase-activated receptor (PAR) 2 is expressed on a subset of primary afferent neurons and involved in inflammatory nociception. Transient receptor potential vanilloid subfamily 1 (TRPV1) is a sensory neuron-specific cation channel that responds to capsaicin, protons, or heat stimulus. Here, we show that TRPV1 is coexpressed with PAR2 but not with PAR1 or PAR3, and that TRPV1 can functionally interact with PAR2. In human embryonic kidney 293 cells expressing TRPV1 and PAR2, PAR2 agonists increased capsaicin- or proton-evoked TRPV1 currents through a PKC-dependent pathway. After application of PAR2 agonists, temperature threshold for TRPV1 activation was reduced from 42 degrees C to well below the body temperature. PAR2-mediated Fos expression in spinal cord was decreased in TRPV1-deficient mice. The functional interaction was also observed in mouse DRG neurons and proved at a behavioral level. These represent a novel mechanism through which trypsin or tryptase released in response to tissue inflammation might trigger the sensation of pain by PAR2 activation. PMID- 15128844 TI - Protease-activated receptor 2 sensitizes the capsaicin receptor transient receptor potential vanilloid receptor 1 to induce hyperalgesia. AB - Inflammatory proteases (mast cell tryptase and trypsins) cleave protease activated receptor 2 (PAR2) on spinal afferent neurons and cause persistent inflammation and hyperalgesia by unknown mechanisms. We determined whether transient receptor potential vanilloid receptor 1 (TRPV1), a cation channel activated by capsaicin, protons, and noxious heat, mediates PAR2-induced hyperalgesia. PAR2 was coexpressed with TRPV1 in small- to medium-diameter neurons of the dorsal root ganglia (DRG), as determined by immunofluorescence. PAR2 agonists increased intracellular [Ca2+] ([Ca2+]i) in these neurons in culture, and PAR2-responsive neurons also responded to the TRPV1 agonist capsaicin, confirming coexpression of PAR2 and TRPV1. PAR2 agonists potentiated capsaicin-induced increases in [Ca2+]i in TRPV1-transfected human embryonic kidney (HEK) cells and DRG neurons and potentiated capsaicin-induced currents in DRG neurons. Inhibitors of phospholipase C and protein kinase C (PKC) suppressed PAR2-induced sensitization of TRPV1-mediated changes in [Ca2+]i and TRPV1 currents. Activation of PAR2 or PKC induced phosphorylation of TRPV1 in HEK cells, suggesting a direct regulation of the channel. Intraplantar injection of a PAR2 agonist caused persistent thermal hyperalgesia that was prevented by antagonism or deletion of TRPV1. Coinjection of nonhyperalgesic doses of PAR2 agonist and capsaicin induced hyperalgesia that was inhibited by deletion of TRPV1 or antagonism of PKC. PAR2 activation also potentiated capsaicin-induced release of substance P and calcitonin gene-related peptide from superfused segments of the dorsal horn of the spinal cord, where they mediate hyperalgesia. We have identified a novel mechanism by which proteases that activate PAR2 sensitize TRPV1 through PKC. Antagonism of PAR2, TRPV1, or PKC may abrogate protease-induced thermal hyperalgesia. PMID- 15128845 TI - Four classes of intercellular channels between glial cells in the CNS. AB - Astrocytes form extensive gap junctions with other astrocytes and with oligodendrocytes. Junctional communication between CNS glia is likely of critical importance because loss of the gap junction channel-forming proteins, connexins Cx32 and Cx47, result in severe demyelination. However, CNS glia express at least six connexins, and the cellular origins and relationships of these proteins have not been determined. We produced a Cx29 reporter mouse in which the connexin coding sequence was replaced with a histological marker, which was used to demonstrate that Cx29, Cx32, and Cx47 are expressed specifically in oligodendrocytes. To determine the relationships between astrocyte and oligodendrocyte connexins, we used double- and triple-immunofluorescence microscopy using semithin sections (<1 microm) of adult mouse spinal cord. Astrocytes form two distinct classes of gap junctions with each other; those composed of Cx26 and those composed of Cx43 and Cx30. In addition, astrocytes establish two classes of intercellular channels with oligodendrocytes, heterotypic Cx26-Cx32 channels and heterotypic Cx30/Cx43-Cx47 channels that may also be heteromeric. In contrast, Cx29 does not colocalize with any of the other five connexins. The data provide the first in vivo demonstration of heterotypic intercellular channels and reveal an unexpected complexity in the composition of glial gap junctions. PMID- 15128846 TI - Mitogen- and stress-activated protein kinase 1 mediates cAMP response element binding protein phosphorylation and activation by neurotrophins. AB - Activation of the transcription factor cAMP response element-binding protein (CREB) by neurotrophins is believed to regulate the survival, differentiation, and maturation of neurons in the CNS and PNS. Although phosphorylation of Ser133 is critical for the expression of CREB-regulated genes, the identity of neurotrophin-regulated Ser133 kinases has remained controversial. We show here that neurotrophin-induced CREB phosphorylation in CNS neurons depends exclusively on the extracellular signal-regulated kinase 1/2-activated kinase mitogen- and stress-activated protein kinase 1 (MSK1). Small interfering RNA directed against ribosomal S6 kinase 1 (RSK1) and RSK2 reduced phosphorylation of a RSK substrate but did not effect CREB-dependent transcription. However, expression of a selective inhibitory MSK1 mutant markedly attenuated BDNF-stimulated CREB phosphorylation and CREB-mediated transcription. Moreover, the ability of neurotrophins to stimulate CREB phosphorylation was abolished in CNS neurons from MSK1 knock-out mice. Consistent with a role for MSK1 in Ser133 phosphorylation, neurotrophin-induced expression of CREB-regulated genes was attenuated in MSK deficient neurons. These results indicate that MSK1 is the major neurotrophin activated Ser133 kinase in CNS neurons. PMID- 15128847 TI - CD4-positive T cell-mediated neuroprotection requires dual compartment antigen presentation. AB - Our laboratory discovered that CD4-positive (CD4+) T cells of the immune system convey transitory neuroprotection to injured mouse facial motoneurons (FMNs) (Serpe et al., 1999, 2000, 2003). A fundamental question in the mechanisms responsible for neuroprotection concerns the identity of the cell(s) that serves as the antigen-presenting cell (APC) to activate the CD4+ T cells. Here, we first establish that CD4+ T cells reactive to non-CNS antigen fail to support FMN survival and, second, demonstrate a two-compartment model of CD4+ T cell activation. Mouse bone marrow (BM) chimeras were developed that discriminate between resident antigen-presenting host cell and BM-derived antigen-presenting donor cell expression of major histocompatibility complex II within central and peripheral compartments, respectively. After facial nerve transection, neither compartment alone is sufficient to result in activated CD4+ T cell-mediated FMN survival. Rather, CD4+ T cell-mediated neuroprotection appears to depend on both resident microglial cells in the central compartment and a BM-derived APC in the peripheral compartment. This is the first in vivo report demonstrating a neuroprotective mechanism requiring APC functions by resident (i.e., parenchymal) microglial cells. PMID- 15128848 TI - Brain-derived neurotrophic factor and trkB signaling in parasympathetic neurons: relevance to regulating alpha7-containing nicotinic receptors and synaptic function. AB - Parasympathetic neurons do not require neurotrophins for survival and are thought to lack high-affinity neurotrophin receptors (i.e., trks). We report here, however, that mRNAs encoding both brain-derived neurotrophic factor (BDNF) and its high-affinity receptor tropomyosin-related kinase B (trkB) are expressed in the parasympathetic chick ciliary ganglion (CG) and that BDNF-like protein is present in the ganglion and in the iris, an important peripheral target of ciliary neurons. Moreover, CG neurons express surface trkB and exogenous BDNF not only initiates trk-dependent signaling, but also alters nicotinic acetylcholine receptor (nAChR) expression and synaptic transmission. In particular, BDNF applied to CG neurons rapidly activates cAMP-dependent response element-binding protein (CREB), and over the long-term selectively upregulates expression of alpha7-subunit-containing, homomeric nAChRs (alpha7-nAChRs), increasing alpha7 subunit mRNA levels, alpha7-nAChR surface sites, and alpha7-nAChR-mediated whole cell currents. At nicotinic synapses formed on CG neurons in culture, brief and long-term BDNF treatments also increase the frequency of spontaneous EPSCs, most of which are mediated by heteromeric nAChRs containing alpha3, alpha5, beta4, and beta2 subunits (alpha3*-nAChRs) with a minor contribution from alpha7-nAChRs. Our findings demonstrate unexpected roles for BDNF-induced, trk-dependent signaling in CG neurons, both in regulating expression of alpha7-nAChRs and in enhancing transmission at alpha3*-nAChR-mediated synapses. The presence of BDNF-like protein in CG and iris target coupled with that of functional trkB on CG neurons raise the possibility that signals generated by endogenous BDNF similarly influence alpha7-nAChRs and nicotinic synapses in vivo. PMID- 15128849 TI - Parallel processing of sensory input by bursts and isolated spikes. AB - Burst firing is commonly observed in many sensory systems and is proposed to transmit information reliably. Although a number of biophysical burst mechanisms have been identified, the relationship between burst dynamics and information transfer is uncertain. Electrosensory pyramidal cells have a well defined backpropagation-dependent burst mechanism. We used in vivo, in vitro, and modeling approaches to investigate pyramidal cell responses to mimics of behaviorally relevant sensory input. We found that within a given spike train, bursts are biased toward low-frequency events while isolated spikes simultaneously code for the entire frequency range. We also demonstrated that burst dynamics are essential for optimal feature detection but are not required for stimulus estimation. We conclude that burst and spike dynamics can segregate a single spike train into two parallel and complementary streams of information transfer. PMID- 15128850 TI - p75 neurotrophin receptor signaling regulates growth cone filopodial dynamics through modulating RhoA activity. AB - The mechanisms by which neurotrophins regulate growth cone motility are unclear. We investigated the role of the p75 neurotrophin receptor (p75NTR) in mediating neurotrophin-induced increases in filopodial length. Our data demonstrate that neurotrophin binding to p75NTR is necessary and sufficient to regulate filopodial dynamics. Furthermore, retinal and dorsal root ganglion growth cones from p75 mutant mice are insensitive to neurotrophins but display enhanced filopodial lengths comparable with neurotrophin-treated wild-type growth cones. This suggests unoccupied p75NTR negatively regulates filopodia length. Furthermore, p75NTR regulates RhoA activity to mediate filopodial dynamics. Constitutively active RhoA blocks neurotrophin-induced increases in filopodial length, whereas inhibition of RhoA enhances filopodial lengths, similar to neurotrophin treatment. BDNF treatment of retinal neurons results in reduced RhoA activity. Furthermore, p75 mutant neurons display reduced levels of activated RhoA compared with wild-type counterparts, consistent with the enhanced filopodial lengths observed on mutant growth cones. These observations suggest that neurotrophins regulate filopodial dynamics by depressing the activation of RhoA that occurs through p75NTR signaling. PMID- 15128851 TI - Memory impairment in aged primates is associated with focal death of cortical neurons and atrophy of subcortical neurons. AB - Mechanisms of cognitive decline with aging remain primarily unknown. We determined whether localized cell loss occurred in brain regions associated with age-related cognitive decline in primates. On a task requiring the prefrontal cortex, aged monkeys were impaired in maintaining representations in working memory. Stereological quantification in area 8A, a prefrontal region associated with working memory, demonstrated a significant 32 +/- 11% reduction in the number of Nissl-stained neurons compared with young monkeys. Furthermore, the number of immunolabeled cholinergic neurons projecting to this region of cortex from the nucleus basalis was also reduced by 50 +/- 6%. In contrast, neuronal number was strikingly preserved in an adjoining prefrontal cortical region also associated with working memory, area 46, and in the component of the nucleus basalis projecting to this region. These findings demonstrate extensive but highly localized loss of neocortical neurons in aged, cognitively impaired monkeys that likely contributes to cognitive decline. Cell degeneration, when present, extends transneuronally. PMID- 15128852 TI - Interplay between local GABAergic interneurons and relay neurons generates gamma oscillations in the rat olfactory bulb. AB - Olfactory stimuli have been known for a long time to elicit oscillations in olfactory brain areas. In the olfactory bulb (OB), odors trigger synchronous oscillatory activity that is believed to arise from the coherent and rhythmic discharges of large numbers of neurons. These oscillations are known to take part in encoding of sensory information before their transfer to higher subcortical and cortical areas. To characterize the cellular mechanisms underlying gamma (30 80 Hz) local field potential (LFP) oscillations, we simultaneously recorded multiunit discharges, intracellular responses, and LFP in rat OB slices. We showed that a single and brief electrical stimulation of olfactory nerve elicited LFP oscillations in the mitral cell body layer lasting >1 sec. Both action potentials and subthreshold oscillations of mitral/tufted cells, the bulbar output neurons, were precisely synchronized with LFP oscillations. This synchronization arises from the interaction between output neurons and granule cells, the main population of local circuit inhibitory interneurons, through dendrodendritic synapses. Interestingly enough, the synchronization exerted by reciprocal synaptic interactions did not require action potentials initiated in granule cell somata. Finally, local application of a GABA(A) receptor antagonist at the mitral cell and external plexiform layers confirmed the exclusive role of the granule cell reciprocal synapses in generating the evoked oscillations. We concluded that interneurons located in the granule cell layer generate synaptic activity capable of synchronizing activity of output neurons by interacting with both their subthreshold and spiking activity. PMID- 15128853 TI - Cannabinoids enhance subsecond dopamine release in the nucleus accumbens of awake rats. AB - Dopaminergic neurotransmission has been highly implicated in the reinforcing properties of many substances of abuse, including marijuana. Cannabinoids activate ventral tegmental area dopaminergic neurons, the main ascending projections of the mesocorticolimbic dopamine system, and change their spiking pattern by increasing the number of impulses in a burst and elevating the frequency of bursts. Although they also increase time-averaged striatal dopamine levels for extended periods of time, little is known about the temporal structure of this change. To elucidate this, fast-scan cyclic voltammetry was used to monitor extracellular dopamine in the nucleus accumbens of freely moving rats with subsecond timescale resolution. Intravenous administration of the central cannabinoid (CB1) receptor agonist, R(+)-[2,3-dihydro-5-methyl-3 [(morpholinyl)methyl]pyrrolo[1,2,3-de]-1,4-benzoxazin-6-yl]-(1-naphthalenyl) methanone mesylate, dose-dependently produced catalepsy, decreased locomotion, and reduced the amplitude of electrically evoked dopamine release while markedly increasing the frequency of detected (nonstimulated) dopamine concentration transients. The CB1 receptor antagonist [N-piperidino-5-(4-chlorophenyl)-1-(2,4 dichlorophenyl)-4-methylpyrazole-3-carboxamide] reversed and prevented all agonist-induced effects but did not show effects on dopamine release when injected alone. These data demonstrate that doses of a cannabinoid agonist known to increase burst firing produce ongoing fluctuations in extracellular dopamine on a previously unrecognized temporal scale in the nucleus accumbens. PMID- 15128854 TI - Variant brain-derived neurotrophic factor (BDNF) (Met66) alters the intracellular trafficking and activity-dependent secretion of wild-type BDNF in neurosecretory cells and cortical neurons. AB - Brain-derived neurotrophic factor (BDNF) plays a critical role in nervous system and cardiovascular development and function. Recently, a common single nucleotide polymorphism in the bdnf gene, resulting in a valine to methionine substitution in the prodomain (BDNF(Met)), has been shown to lead to memory impairment and susceptibility to neuropsychiatric disorders in humans heterozygous for the variant BDNF. When expressed by itself in hippocampal neurons, less BDNF(Met) is secreted in an activity-dependent manner. The nature of the cellular defect when both BDNF(Met) and wild-type BDNF (BDNF(Val)) are present in the same cell is not known. Given that this is the predominant expression profile in humans, we examined the effect of coexpressed BDNF(Met) on BDNF(Val) intracellular trafficking and processing. Our data indicate that abnormal trafficking of BDNF(Met) occurred only in neuronal and neurosecretory cells and that BDNF(Met) could alter the intracellular distribution and activity-dependent secretion of BDNF(Val). We determined that, when coexpressed in the same cell, approximately 70% of the variant BDNF forms BDNF(Val).BDNF(Met) heterodimers, which are inefficiently sorted into secretory granules resulting in a quantitative decreased secretion. Finally, we determined the form of BDNF secreted in an activity-dependent manner and observed no differences in the forms of BDNF(Met) or the BDNF(Val).BDNF(Met) heterodimer compared with BDNF(Val). Together, these findings indicate that components of the regulated secretory machinery interacts specifically with a signal in the BDNF prodomain and that perturbations in BDNF trafficking may lead to selective impairment in CNS function. PMID- 15128855 TI - Glutamate receptor-mediated oligodendrocyte toxicity in periventricular leukomalacia: a protective role for topiramate. AB - Periventricular leukomalacia is a form of hypoxic-ischemic cerebral white matter injury seen most commonly in premature infants and is the major antecedent of cerebral palsy. Glutamate receptor-mediated excitotoxicity is a predominant mechanism of hypoxic-ischemic injury to developing cerebral white matter. We have demonstrated previously the protective effect of AMPA-kainate-type glutamate receptor blockade in a rodent model of periventricular leukomalacia. The present study explores the therapeutic potential of glutamate receptor blockade for hypoxic-ischemic white matter injury. We demonstrate that AMPA receptors are expressed on developing human oligodendrocytes that populate fetal white matter at 23-32 weeks gestation, the period of highest risk for periventricular leukomalacia. We show that the clinically available anticonvulsant topiramate, when administered post-insult in vivo, is protective against selective hypoxic ischemic white matter injury and decreases the subsequent neuromotor deficits. We further demonstrate that topiramate attenuates AMPA-kainate receptor-mediated cell death and calcium influx, as well as kainate-evoked currents in developing oligodendrocytes, similar to the AMPA-kainate receptor antagonist 6-nitro-7 sulfamoylbenzo-(f)quinoxaline-2,3-dione (NBQX). Notably, protective doses of NBQX and topiramate do not affect normal maturation and proliferation of oligodendrocytes either in vivo or in vitro. Taken together, these results suggest that AMPA-kainate receptor blockade may have potential for translation as a therapeutic strategy for periventricular leukomalacia and that the mechanism of protective efficacy of topiramate is caused at least in part by attenuation of excitotoxic injury to premyelinating oligodendrocytes in developing white matter. PMID- 15128856 TI - p35/cyclin-dependent kinase 5 phosphorylation of ras guanine nucleotide releasing factor 2 (RasGRF2) mediates Rac-dependent Extracellular Signal-regulated kinase 1/2 activity, altering RasGRF2 and microtubule-associated protein 1b distribution in neurons. AB - Cyclin-dependent kinase 5 (Cdk5) is a proline-directed kinase the activity of which is dependent on association with its neuron-specific activators, p35 and p39. Cdk5 activity is critical for the proper formation of cortical structures and lamination during development. In the adult nervous system, Cdk5 function is implicated in cellular adhesion, dopamine signaling, neurotransmitter release, and synaptic activity. In addition, Cdk5 is also involved in "cross-talk" with other signal transduction pathways. To further examine its involvement in cross talk with other pathways, we identified proteins that interacted with p35 using the yeast two-hybrid system. We report here that p35 associates with Ras guanine nucleotide releasing factor 2 (RasGRF2) in coimmunoprecipitation and colocalization studies using transfected cell lines as well as primary cortical neurons. Additionally, Cdk5 phosphorylates RasGRF2 both in vitro and in vivo, leading to a decrease in Rac-guanidine exchange factor activity and a subsequent reduction in extracellular signal-regulated kinase 1/2 activity. We show that p35/Cdk5 phosphorylates RasGRF2 on serine737, which leads to an accumulation of RasGRF2 in the neuronal cell bodies coinciding with an accumulation of microtubule-associated protein 1b. The membrane association of p35 and subsequent localization of Cdk5 activity toward RasGRF2 and Rac provide insights into important cellular signaling processes that occur at the membrane, resulting in downstream effects on signal transduction cascades. PMID- 15128857 TI - Conditioning injury-induced spinal axon regeneration fails in interleukin-6 knock out mice. AB - Regeneration of injured adult sensory neurons within the CNS is essentially abortive, attributable in part to lesion-induced or revealed inhibitors such as the chondroitin sulfate proteoglycans and the myelin inhibitors (Nogo-A, MAG, and OMgp). Much of this inhibition may be overcome by boosting the growth status of sensory neurons by delivering a conditioning lesion to their peripheral branches. Here, we identify a key role for the lesion-induced cytokine interleukin-6 (IL-6) in mediating conditioning lesion-induced enhanced regeneration of injured dorsal column afferents. In adult mice, conditioning injury to the sciatic nerve 1 week before bilateral dorsal column crush resulted in regeneration of dorsal column axons up to and beyond the injury site into host CNS tissue. This enhanced growth state was accompanied by an increase in the expression of the growth-associated protein GAP43 in preinjured but not intact dorsal root ganglia (DRGs). Preconditioning injury of the sciatic nerve in IL-6 -/- mice resulted in the total failure in regeneration of dorsal column axons consequent on the lack of GAP43 upregulation after a preconditioning injury. DRGs cell counts and cholera toxin beta subunit labeling revealed that impaired regeneration in knock-out mice was unrelated to cell loss or a deficit in tracer transport. In vitro, exogenous IL-6 boosted sensory neuron growth status as evidenced by enhanced neurite extension. This effect required NGF or NT-3 but not soluble IL-6 receptor as cofactors. Evidence of conditioning lesion-enhanced growth status of DRGs cells can also be observed in vitro as an earlier and enhanced rate of neurite extension; this phenomenon fails in IL-6 -/- mice preinjured 7 d in vivo. We suggest that injury-induced IL-6 upregulation is required to promote regeneration within the CNS. Our results indicate that this is achieved through a boosted growth state of dorsal column projecting sensory neurons. PMID- 15128858 TI - Transient receptor potential vanilloid 4 is essential in chemotherapy-induced neuropathic pain in the rat. AB - The development of treatments for neuropathic pain has been hindered by our limited understanding of the basic mechanisms underlying abnormalities in nociceptor hyperexcitability. We recently showed that the polymodal receptor transient receptor potential vanilloid 4 (TRPV4), a member of the transient receptor potential (TRP) family of ion channels, may play a role in inflammatory pain (Alessandri-Haber et al., 2003). The present study tested whether TRVP4 also contributes to neuropathic pain, using a rat model of Taxol-induced painful peripheral neuropathy. Taxol is the most widely used drug for the treatment of a variety of tumor types, but the dose of Taxol that can be tolerated is limited by the development of a small-fiber painful peripheral neuropathy. We found that Taxol treatment enhanced the nociceptive behavioral responses to both mechanical and hypotonic stimulation of the hind paw. Spinal administration of antisense oligodeoxynucleotides to TRPV4, which reduced the expression of TRPV4 in sensory nerve, abolished Taxol-induced mechanical hyperalgesia and attenuated hypotonic hyperalgesia by 42%. The enhancement of osmotic nociception involves sensitization of osmotransduction in primary afferents because osmotransduction was enhanced in cultured sensory neurons isolated from Taxol-treated rats. Taxol induced TRPV4-mediated hyperalgesia and the enhanced osmotransduction in cultured nociceptors were dependent on integrin/Src tyrosine kinase signaling. These results suggest that TRPV4 plays a crucial role in a painful peripheral neuropathy, making it a very promising target for the development of a novel class of analgesics. PMID- 15128859 TI - Behavioral context and coherent oscillations in the supplementary motor area. AB - Movements with similar physical characteristics can occur in various behavioral contexts, as when they are embedded in different sequences or when the expected outcomes of movements vary. Similarly, neurons in various sensory and motor structures in the brain commonly display modulations in their activity according to contextual factors, such as expected reward. Although these contextual signals must be combined with incoming sensory inputs to generate appropriate behaviors according to the animal's motivational state, the mechanisms by which these two signals are integrated remain poorly understood. The present study examined the effects of contextual factors on the magnitude of coherent oscillations in the activity of individual neurons recorded in the supplementary motor area (SMA) of monkeys during a serial reaction time task. In this task, the animal produced a predictable sequence of hand movements repeatedly according to visual instructions. The performance of the animal was influenced by the location of the rewarded target as well as the ordinal position of the movement. In contrast, the level of coherent oscillations in the activity of SMA neurons was affected only by the rewarded target location but not by the ordinal position of the movement sequence. In addition, changes in coherent oscillations were not accounted for by systematic changes in the mean firing rates. These results are consistent with the proposal that synchronous spikes might be used to control the flow of information and suggest that coherent oscillations in the SMA might encode contextual variables, such as expected reward. PMID- 15128860 TI - Learning at different satiation levels reveals parallel functions for the cAMP protein kinase A cascade in formation of long-term memory. AB - Learning and memory formation in intact animals is generally studied under defined parameters, including the control of feeding. We used associative olfactory conditioning of the proboscis extension response in honeybees to address effects of feeding status on processes of learning and memory formation. Comparing groups of animals with different but defined feeding status at the time of conditioning reveals new and characteristic features in memory formation. In animals fed 18 hr earlier, three-trial conditioning induces a stable memory that consists of different phases: a mid-term memory (MTM), translation-dependent early long-term memory (eLTM; 1-2 d), and a transcription-dependent late LTM (lLTM; > or =3 d). Additional feeding of a small amount of sucrose 4 hr before conditioning leads to a loss of all of these memory phases. Interestingly, the basal activity of the cAMP-dependent protein kinase A (PKA), a key player in LTM formation, differs in animals with different satiation levels. Pharmacological rescue of the low basal PKA activity in animals fed 4 hr before conditioning points to a specific function of cAMP-PKA cascade in mediating satiation dependent memory formation. An increase in PKA activity during conditioning rescues only transcription-dependent lLTM; acquisition, MTM, and eLTM are still impaired. Thus, during conditioning, the cAMP-PKA cascade mediates the induction of the transcription-dependent lLTM, depending on the satiation level. This result provides the first evidence for a central and distinct function of the cAMP-PKA cascade connecting satiation level with learning. PMID- 15128861 TI - Expression of a poly-glutamine-ataxin-3 transgene in orexin neurons induces narcolepsy-cataplexy in the rat. AB - The sleep disorder narcolepsy has been linked to loss of hypothalamic neurons producing the orexin (hypocretin) neuropeptides. Here, we report the generation of transgenic rats expressing a human ataxin-3 fragment with an elongated polyglutamyl stretch under control of the human prepro-orexin promoter (orexin/ataxin-3 rats). At 17 weeks of age, the transgenic rats exhibited postnatal loss of orexin-positive neurons in the lateral hypothalamus, and orexin containing projections were essentially undetectable. The loss of orexin production resulted in the expression of a phenotype with fragmented vigilance states, a decreased latency to rapid eye movement (REM) sleep and increased REM sleep time during the dark active phase. Wakefulness time was also reduced during the dark phase, and this effect was concentrated at the photoperiod boundaries. Direct transitions from wakefulness to REM sleep, a defining characteristic of narcolepsy, occurred frequently. Brief episodes of muscle atonia and postural collapse resembling cataplexy were also noted while rats maintained the electroencephalographic characteristics of wakefulness. These findings indicate that the orexin/ataxin-3 transgenic rat could provide a useful model of human narcolepsy. PMID- 15128862 TI - Changes in intracellular chloride after oxygen-glucose deprivation of the adult hippocampal slice: effect of diazepam. AB - Ischemic injury to the CNS results in loss of ionic homeostasis and the development of neuronal death. An increase in intracellular Ca2+ is well established, but there are few studies of changes in intracellular Cl- ([Cl-]i) after ischemia. We used an in vitro model of cerebral ischemia (oxygen-glucose deprivation) to examine changes in [Cl-]i and GABA(A) receptor-mediated responses in hippocampal slices from adult rats. Changes in [Cl-]i were measured in area CA1 pyramidal neurons using optical imaging of 6-methoxy-N-ethylquinolinium chloride, a Cl--sensitive fluorescent indicator. Oxygen-glucose deprivation induced an immediate rise in [Cl-]i, which recovered within 20 min. A second and more prolonged rise in [Cl-]i occurred within the next hour, during which postsynaptic field potentials failed to recover. The sustained increase in [Cl-]i was not blocked by GABA(A) receptor antagonists. However, oxygen-glucose deprivation caused a progressive downregulation of the K+-Cl- cotransporter (KCC2), which may have contributed to the Cl- accumulation. The rise in [Cl-]i was accompanied by an inability of the GABA(A) agonist muscimol to cause Cl- influx. In vivo, diazepam is neuroprotective when given early after ischemia, although the mechanism by which this occurs is not well understood. Here, we added diazepam early after oxygen-glucose deprivation and prevented the downregulation of KCC2 and the accumulation of [Cl-]i. Consequently, both GABA(A) responses and synaptic transmission within the hippocampus were restored. Thus, after oxygen-glucose deprivation, diazepam may decrease neuronal excitability, thereby reducing the energy demands of the neuron. This may prevent the activation of downstream cell death mechanisms and restore Cl- homeostasis and neuronal function PMID- 15128863 TI - Enhanced anti-inflammatory potency of a nitric oxide-releasing derivative of flunisolide: role of nuclear factor-kappaB. AB - Glucocorticoids remain among the most commonly used anti-inflammatory drugs, despite significant adverse effects. Other anti-inflammatory drugs, including aspirin, have been coupled through an ester linkage to a nitric oxide-releasing moiety, resulting in an increase in potency and a decrease in adverse effects. Prednisolone has similarly been modified, with marked improvement of its therapeutic index. In the present study, we have evaluated whether a nitric oxide releasing derivative of another glucocorticoid, flunisolide, would increase its potency as an anti-inflammatory agent and would decrease its systemic toxicity. To evaluate anti-inflammatory potency and efficacy, the carrageenan-airpouch model in the rat was used. Flunisolide and NCX-1024 (flunisolide-21-[4' (nitrooxymethyl) benzoate]) were compared across a range of doses, with both direct injection into the airpouch and oral administration. The ability of these agents to protect the stomach against indomethacin-induced damage also was assessed. Effects of oral administration of the two drugs on body weight gain and adrenal suppression were also evaluated. With direct application into the airpouch, NCX-1024 was found to be 41 times more potent than flunisolide in reducing leukocyte accumulation and prostaglandin E2 generation. The increased potency may be related to an enhanced ability of NCX-1024 to prevent nuclear factor-kappaB activation. When given orally, the two compounds exhibited similar potency. However, orally administered NCX-1024 was more potent at protecting against indomethacin-induced gastric damage, caused less reduction of body weight, and, unlike flunisolide, did not cause adrenal atrophy. These studies suggest that NCX-1024 may be an attractive alternative to conventional glucocorticoids, particularly for applications involving topical administration. PMID- 15128864 TI - Disposition of flavonoids via enteric recycling: enzyme-transporter coupling affects metabolism of biochanin A and formononetin and excretion of their phase II conjugates. AB - The purpose of this study was to continue our effort to determine how enzyme transporter coupling affect disposition of flavonoids. The rat intestinal perfusion and Caco-2 cell models were used together with relevant microsomes. In perfusion model, isoflavone (i.e., formononetin and biochanin A) absorption and subsequent excretion of its metabolites were always site-dependent. Maximal amounts of intestinal and biliary conjugates excreted per 30 min were 31 and 51 nmol for formononetin, more than that for pure biochanin A (12 and 20 nmol). When a standardized red clover extract (biochanin A/formononetin = 10:7) was used, the results indicated that more metabolites of biochanin A than formononetin were found in the perfusate (36.9 versus 22.8 nmol) and bile (78 versus 51 nmol). In metabolism studies, rat intestinal and liver microsomes always glucuronidated biochanin A faster (p < 0.05) than formononetin, whereas intestinal microsomes glucuronidated both isoflavones faster (p < 0.05) than liver microsomes. However, rapid metabolism in the microsomes did not translate into more efficient excretion in either the rat perfusion model as shown previously or in the Caco-2 model. In the Caco-2 model, both isoflavones were rapidly absorbed, efficiently conjugated, and the conjugates excreted apically and basolaterally. More formononetin conjugates were excreted than biochanin A when used alone, but much more biochanin A conjugates were found when using the isoflavone mixture. In conclusion, efficiency of enzyme-transporter coupling controls the amounts of metabolites excreted by the intestine and liver and determines the relative contribution of enteric and enterohepatic recycling to the in vivo disposition of isoflavones. PMID- 15128865 TI - Protein cross-linkage induced by formaldehyde derived from semicarbazide sensitive amine oxidase-mediated deamination of methylamine. AB - Semicarbazide-sensitive amine oxidase (SSAO) catalyzes the conversion of methylamine to formaldehyde. This enzyme is located on the surface of the cytoplasmic membrane and in the cytosol of vascular endothelial cells, smooth muscle cells, and adipocytes. Increased SSAO activity has been found in patients with diabetes mellitus, chronic heart failure, and multiple types of cerebral infarcts and is associated with obesity. Increased SSAO-mediated deamination may contribute to protein deposition, the formation of plaques, and inflammation, and thus may be involved in the pathophysiology of chronic vascular and neurological disorders, such as diabetic complications, atherosclerosis, and Alzheimer's disease. In the present study, we demonstrate the induction of cross-linkage of formaldehyde with the lysine moiety of peptides and proteins. Formaldehyde protein adducts were reduced with sodium cyanoborohydride, hydrolyzed in hydrochloric acid, and the amino acids in the hydrolysates were derivatized with fluorenylmethyl chloroformate and then identified with high-performance liquid chromatography. We further demonstrate that incubation of methylamine in the presence of SSAO-rich tissues, e.g., human brain meninges, results in formaldehyde-protein cross-linkage of particulate bound proteins as well as of soluble proteins. This cross-linkage can be completely blocked by a selective inhibitor of SSAO. Our data support the hypothesis that the SSAO-induced production of formaldehyde may be involved in the alteration of protein structure, which may subsequently cause protein deposition associated with chronic pathological disorders. PMID- 15128866 TI - Analysis of microtubule sliding patterns in Chlamydomonas flagellar axonemes reveals dynein activity on specific doublet microtubules. AB - Generating the complex waveforms characteristic of beating eukaryotic cilia and flagella requires spatial regulation of dynein-driven microtubule sliding. To generate bending, one prediction is that dynein arms alternate between active and inactive forms on specific subsets of doublet microtubules. Using an in vitro microtubule sliding assay combined with a structural approach, we determined that ATP induces sliding between specific subsets of doublet microtubules, apparently capturing one phase of the beat cycle. These studies were also conducted using high Ca2+ conditions. In Chlamydomonas, high Ca2+ induces changes in waveform which are predicted to result from regulating dynein activity on specific microtubules. Our results demonstrate that microtubule sliding in high Ca2+ buffer is also induced by dynein arms on specific doublets. However, the pattern of microtubule sliding in high Ca2+ buffer significantly differs from that in low Ca2+. These results are consistent with a 'switching hypothesis' of axonemal bending and provide evidence to indicate that Ca2+ control of waveform includes modulation of the pattern of microtubule sliding between specific doublets. In addition, analysis of microtubule sliding in mutant axonemes reveals that the control mechanism is disrupted in some mutants. PMID- 15128867 TI - Connexin43 and connexin26 form gap junctions, but not heteromeric channels in co expressing cells. AB - Many cells contain two (or more) gap junction proteins that are able to oligomerize with each other to form heteromeric gap junction channels and influence the properties of intercellular communication. Cx26 and Cx43 are found together in a number of cell types, but previous data have suggested that they might not form heteromeric connexons. We studied the possible interactions of these connexins by co-expression in three different cell lines. Analysis of N2aCx26/Cx43 cell pairs by double whole-cell patch-clamp methods showed that these cells were coupled, but contained only a small number of sizes of single channels consistent with those formed by homomeric Cx26 or Cx43 channels. Immunofluorescence studies showed that both connexins localized to appositional membranes, but in largely distinct domains. Analysis of Triton X-100-solubilized connexons from co-expressing cells by centrifugation through sucrose gradients or by affinity purification using a Ni-NTA column showed no evidence of mixing of Cx26 and Cx43. These results contrast with our observations in cells co expressing other connexins with Cx43 and suggest that Cx26 and Cx43 do not form heteromeric hemichannels. Moreover, the incorporation of Cx26 and Cx43 into oligomers and into the membrane were similarly affected by treatment of co expressing cells with brefeldin A or nocodazole, suggesting that the lack of mixing is due to incompatibility of these connexins, not to differences in biosynthetic trafficking. PMID- 15128868 TI - Actin- and protein-4.1-containing filaments link nuclear pore complexes to subnuclear organelles in Xenopus oocyte nuclei. AB - We imaged the interiors of relatively intact Xenopus oocyte nuclei by field emission scanning electron microscopy (feSEM) and visualized a network of filaments that attach to nuclear pore complexes and extend throughout the nucleus. Within the nucleus, these 'pore-linked filaments' (PLFs) were embedded into spherical structures 100 nm to approximately 5 microm in diameter. A subset of spheres was identified as Cajal bodies by immuno-gold labeling; the rest were inferred to be nucleoli and snurposomes both of which are abundant in Xenopus oocyte nuclei. Most PLFs were independent of chromatin. The thickness of a typical PLF was 40 nm (range, approximately 12-100 nm), including the 4 nm chromium coat. PLFs located inside the nucleus merged, bundled and forked, suggesting architectural adaptability. The PLF network collapsed upon treatment with latrunculin A, which depolymerizes actin filaments. Jasplakinolide, which stabilizes actin filaments, produced PLFs with more open substructure including individual filaments with evenly-spaced rows of radially projecting short filaments. Immuno-gold labeling of untreated oocyte nuclei showed that actin and protein 4.1 each localized on PLFs. Protein 4.1-gold epitopes were spaced at approximately 120 nm intervals along filaments, and were often paired ( approximately 70 nm apart) at filament junctions. We suggest that protein 4.1 and actin contribute to the structure of a network of heterogeneous filaments that link nuclear pore complexes to subnuclear organelles, and discuss possible functions for PLFs in nuclear assembly and intranuclear traffic. PMID- 15128869 TI - Expression of fragments of translation initiation factor eIF4GI reveals a nuclear localisation signal within the N-terminal apoptotic cleavage fragment N-FAG. AB - The eukaryotic initiation factor eIF4GI plays a central role in the assembly of a competent initiation complex at the 5' end of an mRNA. Five isoforms of eIF4G exist in cells, arising from alternative translation initiation. During picornaviral infection or apoptosis, eIF4GI is cleaved proteolytically to yield distinct fragments. Using HeLa cells, we have examined the fate of these proteins in the cell. We have found that while endogenous eIF4GI is predominantly cytoplasmic, a population can also be visualised in the nucleus. Furthermore, eIF4GI is localised primarily at the nuclear periphery in the vicinity of eIF4E and PABP1. Transient transfection of HeLa cells with different myc-tagged isoforms of eIF4GI did not result in any obvious differences in their localisation. However, expression of discrete fragments of eIF4GI corresponding to those generated after apoptosis or picornaviral infection generated a distinctive, but intricate localisation pattern. Our work shows that the N terminal apoptotic cleavage fragment N-FAG contains a sequence of basic amino acids that can act as a nuclear localisation signal. In addition, the presence or absence of the sequence flanking and including the eIF4E binding site (residues 533-682) confers a distinct cellular distribution pattern for the central domain of eIF4GI. PMID- 15128870 TI - A role for the Cdc14-family phosphatase Flp1p at the end of the cell cycle in controlling the rapid degradation of the mitotic inducer Cdc25p in fission yeast. AB - The Schizosaccaromyces pombe protein Flp1p belongs to a conserved family of serine-threonine-phosphatases. The founding member of this family, Saccharomyces cerevisiae Cdc14p, is required for inactivation of mitotic CDKs and reversal of CDK mediated phosphorylation at the end of mitosis, thereby bringing about the M G1 transition. Initial studies of Flp1p suggest that it may play a different role to Cdc14p. Here we show that Flp1p is required for rapid degradation of the mitotic inducer Cdc25p at the end of mitosis, and that Cdc25p is a substrate of Flp1p in vitro. Down-regulation of Cdc25p activity by Flp1p may ensure a prompt inactivation of mitotic CDK complexes to trigger cell division. Our results suggest a regulatory mechanism, and a universal role, for Cdc14p like proteins in coordination of cytokinesis with other cell cycle events. PMID- 15128871 TI - Phosphorylation of CDC25B by Aurora-A at the centrosome contributes to the G2-M transition. AB - Aurora-A protein kinase, which is the product of an oncogene, is required for the assembly of a functional mitotic apparatus and the regulation of cell ploidy. Overexpression of Aurora-A in tumour cells has been correlated with cancer susceptibility and poor prognosis. Aurora-A activity is required for the recruitment of CDK1-cyclin B1 to the centrosome prior to its activation and the commitment of the cell to mitosis. In this report, we demonstrate that the CDC25B phosphatase, an activator of cyclin dependent kinases at mitosis, is phosphorylated both in vitro and in vivo by Aurora-A on serine 353 and that this phosphorylated form of CDC25B is located at the centrosome during mitosis. Knockdown experiments by RNAi confirm that the centrosome phosphorylation of CDC25B on S353 depends on Aurora-A kinase. Microinjection of antibodies against phosphorylated S353 results in a mitotic delay whilst overexpression of a S353 phosphomimetic mutant enhances the mitotic inducing effect of CDC25B. Our results demonstrate that Aurora-A phosphorylates CDC25B in vivo at the centrosome during mitosis. This phosphorylation might locally participate in the control of the onset of mitosis. These findings re-emphasise the role of the centrosome as a functional integrator of the pathways contributing to the triggering of mitosis. PMID- 15128872 TI - Control of actin dynamics by p38 MAP kinase - Hsp27 distribution in the lamellipodium of smooth muscle cells. AB - We investigated the role of the p38 mitogen-activated protein kinase (p38 MAPK) in the PDGF-BB-induced cytoskeleton remodeling that occurs during the migration of porcine aortic smooth muscle cells (SMC). We showed that p38 MAPK controlled the polymerization of actin that is required for PDGF-induced lamellipodia formation and migration. To investigate the mechanism of action of p38 MAPK, we explored its cellular localization and that of its indirect substrate, the heat shock protein Hsp27, during SMC spreading on fibronectin in the presence and absence of PDGF. Spreading of SMC on fibronectin activated p38 MAPK in a sustained manner only in the presence of PDGF. In these conditions, Hsp27 and p38 MAPK were localized all over the lamellipodia. A transiently phosphorylated form of p38 MAPK was observed at the leading edge, whereas p38 MAPK remained phosphorylated at the base of the lamellipodia. Phosphorylated Hsp27 was excluded from the leading edge and restricted to the base of the lamellipodia. These results were confirmed by Triton X-100 extraction of particulate membrane fraction. Displacement of Hsp27 from the leading edge by cytochalasin D treatment suggests that nonphosphorylated Hsp27 caps barbed ends in vivo. Our data indicate that nonphosphorylated Hsp27 might contribute to the formation of a short, branched actin network at the leading edge, whereas phosphorylated Hsp27 might stabilize the actin network at the base of lamellipodia, which is composed of long, unbranched actin filaments. PMID- 15128873 TI - Recruitment of Pyk2 and Cbl to lipid rafts mediates signals important for actin reorganization in growing neurites. AB - Protein tyrosine kinase Pyk2 and multifunctional adaptor protein Cbl are implicated in the regulation of the cytoskeleton in several cell types. We report that Pyk2 and Cbl form a signaling complex that is translocated to lipid rafts and is enriched in growth cones of differentiating PC12 cells following growth factor stimulation. We found that Pyk2 and Cbl interacted with the adaptor protein ArgBP2, which also bound to flotillin-1, a component of lipid raft microdomains. These interactions contributed to recruitment of the Pyk2/Cbl complex to lipid raft compartments. In addition, Pyk2, Cbl and ArgBP2 were found co-localized with actin in axons and growth cones of differentiated PC12 cells. Moreover, co-expression of Pyk2, ArgBP2 and Cbl facilitated growth factor-induced formation of lamellipodia at the tip of neurites. Formation of these growth cone lamellipodia was dependent on intact lipid rafts and the Cbl-associated effectors Crk and phosphatidylinositol 3 (PI 3)-kinase. Our results indicate that recruitment of Pyk2/Cbl complexes to lipid rafts participates in growth factor induced regulation of the actin cytoskeleton in growing neurites. PMID- 15128874 TI - Heterochromatin and tri-methylated lysine 20 of histone H4 in animals. AB - Tri-methylated lysine 20 on histone H4 (Me(3)K20H4) is a marker of constitutive heterochromatin in murine interphase and metaphase cells. Heterochromatin marked by Me(3)K20H4 replicates late during S phase of the cell cycle. Serum starvation increases the number of cells that exhibit high levels of Me(3)K20H4 at constitutive heterochromatin. Me(3)K20H4 is also present at the centromeric heterochromatin of most meiotic chromosomes during spermatogenesis and at the pseudoautosomal region, as well as at some telomeres. It is not present on the XY body. During murine embryogenesis the maternal pronucleus contains Me(3)K20H4; Me(3)K20H4 is absent from the paternal pronucleus. On Drosophila polytene chromosomes Me(3)K20H4 is present in a 'punctate pattern' at many chromosomal bands, including the chromocenter. In coccids it is present on the facultatively heterochromatinised paternal chromosome set. We also present evidence that Me(3)K20H4 is dependent upon H3-specific Suv(3)9 histone methyltransferase activity, suggesting that there may be 'epigenetic cross-talk' between histones H3 and H4. PMID- 15128875 TI - The phylogenetic relationship of tetrapod, coelacanth, and lungfish revealed by the sequences of forty-four nuclear genes. AB - The origin of tetrapods is a major outstanding issue in vertebrate phylogeny. Each of the three possible principal hypotheses (coelacanth, lungfish, or neither being the sister group of tetrapods) has found support in different sets of data. In an attempt to resolve the controversy, sequences of 44 nuclear genes encoding amino acid residues at 10,404 positions were obtained and analyzed. However, this large set of sequences did not support conclusively one of the three hypotheses. Apparently, the coelacanth, lungfish, and tetrapod lineages diverged within such a short time interval that at this level of analysis, their relationships appear to be an irresolvable trichotomy. PMID- 15128876 TI - The northeast Indian passageway: a barrier or corridor for human migrations? AB - The northeast Indian passageway connecting the Indian subcontinent to East/Southeast Asia is thought to have been a major corridor for human migrations. Because it is also an important linguistic contact zone, it is predicted that northeast India has witnessed extensive population interactions, thus, leading to high genetic diversity within groups and heterogeneity among groups. To test this prediction, we analyzed 14 biallelic and five short tandem repeat Y-chromosome markers and hypervariable region 1 mtDNA sequence variation in 192 northeast Indians. We find that both northeast Indian Y chromosomes and mtDNAs consistently show strikingly high homogeneity among groups and strong affinities to East Asian groups. We detect virtually no Y-chromosome and mtDNA admixture between northeast and other Indian groups. Northeast Indian groups are also characterized by a greatly reduced Y-chromosome diversity, which contrasts with extensive mtDNA diversity. This is best explained by a male founder effect during the colonization of northeast India that is estimated to have occurred within the past 4,000 years. Thus, contrary to the prediction, these results provide strong evidence for a genetic discontinuity between northeast Indian groups and other Indian groups. We, therefore, conclude that the northeast Indian passage way acted as a geographic barrier rather than as a corridor for human migrations between the Indian subcontinent and East/Southeast Asia, at least within the past millennia and possibly for several tens of thousand years, as suggested by the overall distinctiveness of the Indian and East Asian Y chromosome and mtDNA gene pools. PMID- 15128877 TI - High prevalence of Chlamydia pneumoniae infection in cyclosporin A-induced post transplant gingival overgrowth tissue and evidence for the possibility of persistent infection despite short-term treatment with azithromycin. AB - BACKGROUND: Cyclosporin A (CsA) induces gingival overgrowth (GO) in up to a quarter of CsA-treated renal transplant recipients. A short-term therapy with azithromycin effectively reduces GO, indicating a possible involvement of microorganisms in the pathogenesis of CsA-induced GO. We aimed to determine if there could be any relationship between infection with Chlamydia pneumoniae and GO pathogenesis. In addition, we determined the long-term persistence rate of C. pneumoniae infection in residual GO tissue when azithromycin treatment failed to eliminate GO. METHODS: Chlamydia pneumoniae IgG and IgM antibody titres were measured by microimmunofluorescence technique in sera of kidney recipients with (n = 11) and without (n = 89) GO. GOs were rated and gingivectomies were performed before treatment with 500 mg of azithromycin for 3 days and at months 6 and 12 post-treatment when C. pneumoniae titres were re-evaluated. Nested polymerase chain reaction was performed to identify C. pneumoniae-specific DNA in GO tissues. Results of C. pneumoniae antibody titres from patients with GO were compared with pair-matched controls without GO. RESULTS: Chlamydia pneumoniae IgM titres were elevated in five of 11 patients with GO and in none without GO, whereas the difference of C. pneumoniae IgG titres between patients with GO and pair-matched controls did not reach significance (P<0.57). Chlamydia pneumoniae specific DNA was found in 10 of 11 GO tissue samples pre-treatment. Azithromycin therapy effectively reduced GO and C. pneumoniae IgM titres. In residual GO, C. pneumoniae-specific DNA remained detectable after 1 year in all GO tissue samples despite azithromycin treatment. The C.pneumoniae IgM titres correlated with GO scores. CONCLUSION: Chlamydia pneumoniae infection is highly prevalent in CsA induced GO. The infection can persist over a long period in residual GO despite short-term azithromycin therapy. The results indicate that CsA immunosuppression enhances C. pneumoniae infection rates in non-cardiovascular tissue. PMID- 15128878 TI - Evidence for protective roles of polyethylene glycol plus high sodium solution and trimetazidine against consequences of renal medulla ischaemia during cold preservation and reperfusion in a pig kidney model. AB - BACKGROUND: The renal medulla is particularly sensitive to oxidant stress and to ischaemia-reperfusion injury (IRI). In organ transplantation, delayed graft function is an important problem and cold ischaemia is thought to be the most important factor in short- and long-term complications. Our aim was to study cold induced damage in proximal tubular segments and renal medulla osmolite excretion during use of various preservation solutions, and to clarify the role of trimetazidine (TMZ) in limiting renal dysfunction. METHODS: Using an autotransplanted pig kidney model, we assessed renal tubule function, medullary osmolite excretion and renal damage between day 1 and week 2 after 24 or 48 h cold storage in University of Wisconsin solution (UW), Celsior and ECPEG (two new high Na(+) preservation solutions) or the Hopital Edouard Herriot solution (HEH; a high Na(+) version of UW). In additional groups, TMZ was added to these preservation solutions for 24 and 48 h cold storage. RESULTS: Renal function was reduced under these preservation conditions. Tubular injury was associated with aminoaciduria and with a limited Na(+) reabsorbtion. Medullary damage led to the early appearance of trimethylamine-N-oxide and dimethylamine in urine. However, renal damage was modulated by preservation conditions. In addition, TMZ added to each of the solutions efficiently protected against IRI even after prolonged preservation. CONCLUSION: TMZ efficiently protected kidneys against damage when added to the HEH and particularly ECPEG solutions, even after 24 h cold storage. These findings point to a role for drugs that target mitochondria, and demonstrate that TMZ may provide a valuable therapeutic tool against IRI and could be included in therapeutic protocols. PMID- 15128879 TI - Urinary L-lactate excretion is increased in renal Fanconi syndrome. AB - BACKGROUND: Measurement of l-lactate in body fluids is an established clinical tool to identify disorders of cellular respiration. However, there is very little known about the clinical value of urinary lactate measurements. We investigated urinary lactate excretion in children with renal Fanconi syndrome. METHODS: Freshly voided urine samples were obtained from children with Fanconi syndrome and controls both with and without renal disease. Urine lactate was estimated by conversion to pyruvate in the presence of lactate dehydrogenase and NAD. The NADH produced was measured photometrically. Urine lactate was factored for urine creatinine. RESULTS: Children with Fanconi syndrome had a significantly higher urine lactate/creatinine ratio [mean: 84 x 10(-2) mmol/mmol; 95% confidence interval (CI): 40.8-127.1 x 10(-2) mmol/mmol] than healthy controls (mean: 1.3 x 10(-2) mmol/mmol; CI: 1.1-1.5 x 10(-2) mmol/ mmol) and those with a variety of renal diseases (mean: 3.1 x 10(-2) mmol/mmol; CI: 1.8-4.5 x 10(-2) mmol/mmol). CONCLUSIONS: Urinary lactate is increased in Fanconi syndrome. The increase is likely to be due to reduced lactate co-transport in the proximal tubule. Urinary lactate/creatinine has clinical utility as a sensitive test of disordered proximal renal tubular function. PMID- 15128880 TI - Antifibrotic, nephroprotective potential of ACE inhibitor vs AT1 antagonist in a murine model of renal fibrosis. AB - BACKGROUND: Several studies have shown antifibrotic effects of angiotensin converting enzyme (ACE) inhibitors as well as of angiotensin receptor 1 (AT1) antagonists, however, prospective trials with clinical end points comparing these effects do not exist. COL4A3-/- mice develop a non-hypertensive progressive renal fibrosis. We used this animal model to compare the potential of ACE inhibitor vs AT1 antagonist to prevent renal fibrosis irrespective of blood pressure-dependent involvement by the renin system. METHODS: COL4A3-/- mice were treated with placebo, ramipril or candesartan. Blood pressure, proteinuria, serum urea and lifespan were monitored. Renal matrix was characterized by immuno-histochemistry, light and electron microscopy. Further biochemical analysis was provided using cDNA microarray and western blot techniques. RESULTS: Untreated mice died of renal failure after 71+/-6 days. Ramipril and candesartan both delayed onset and reduced the extent of proteinuria. Both had minor effects on blood pressure and postponed onset of uraemia. Ramipril increased lifespan by 111% to 150+/-21 days (P<0.01), whereas candesartan resulted in only a 38% prolongation to 98+/-16 days (P<0.01). Ramipril reduced glomerular and tubulo-interstitial fibrosis and numbers of activated fibroblasts to a greater extent than candesartan. Microarray and western blot analysis revealed a higher antifibrotic potential of ramipril in terms of downregulation of TGFbeta, connective tissue growth factor, metalloproteinases and extracellular matrix proteins. CONCLUSIONS: The results indicate an antifibrotic, nephroprotective effect of ACE inhibitors and AT1 antagonists in an animal model of progressive renal fibrosis. The greater antifibrotic effect of ramipril at the maximal therapeutic doses employed may not be explained by different antiproteinuric or blood pressure lowering properties, but by-in contrast to candesartan-its ability to hinder the proinflammatory, profibrotic activation of the angiotensin receptor 2. PMID- 15128881 TI - Diagnostic utility and safety of transjugular kidney biopsy in the obese patient. AB - BACKGROUND: The obese patient may be potentially at high risk for traditional percutaneous ultrasound-guided biopsy. The utility of transjugular kidney biopsy (TJKB) in this group of patients has not been established. METHODS: We conducted a retrospective analysis of 37 obese patients who underwent TJKB at our centre. The kidney was approached via the right renal vein in 31 patients. Under fluoroscopy, core biopsies were obtained from the lower pole with a 19G biopsy needle. Post-procedure venogram was performed to assess for contrast extravasation indicating capsule perforation or communication with the collecting system. Patients were followed for procedure-related complications. Mean weight was 128 kg (range: 77-187 kg) and body mass index was 44 kg/m(2) (range: 34-64 kg/m(2)). Mean creatinine was 2.2 mg/dl (range: 0.5-6.5 mg/dl). Fifteen patients had diabetes, five of whom were nephrotic; 10 other patients had nephrotic range proteinuria. RESULTS: Of the 37 patients, six were hospitalized at the time of biopsy and three were admitted for observation. All patients returned to baseline activity the day following procedure. Histopathological diagnosis was made in 33 cases (89.2%) with a mean of 19.2 glomeruli (range: 0-62 glomeruli) per patient. There was one major complication: a delayed retroperitoneal bleed requiring multiple transfusions. Contrast extravasation outside the capsule occurred in five patients and extravasation into the collecting system occurred in three. Body mass index was not associated with number of glomeruli obtained or complication rate. CONCLUSIONS: TJKB in obese patients is a relatively safe, reliable and minimally invasive procedure with an excellent diagnostic yield. PMID- 15128882 TI - Renal effects of N-acetylcysteine in patients at risk for contrast nephropathy: decrease in oxidant stress-mediated renal tubular injury. AB - BACKGROUND: N-Acetylcysteine has been shown to protect against contrast nephropathy, although the mechanisms underlying such an effect are unclear. Surprisingly, studies have shown that post-radiocontrast renal function actually improves in chronic renal failure patients receiving N-acetylcysteine. However, there have been no studies investigating the cause of this improvement. METHODS: In a double-blind, placebo-controlled study, 24 patients (aged 65+/-2 years) suffering from stable mild-to-moderate renal insufficiency and undergoing elective coronary angiography were randomized to receive either placebo or N acetylcysteine. All received similar hydration. Renal function parameters were assessed 48 h before and 48 h after radiocontrast administration. Urinary 15 isoprostane F2(t), a specific marker of oxidative stress, was measured immediately before and after the procedure. Expression of urinary alpha glutathione S-transferase protein, a specific proximal tubular injury marker, was assessed after the procedure. RESULTS: Comparing creatinine clearance values before and after angiography, a significant increase was seen in N-acetylcysteine patients (44.7+/-4.2 vs 57.2+/-6.3 ml/min/1.73 m(2); P = 0.02), whereas placebo patients presented no change (46.6+/-5.0 vs 46.9+/-4.3 ml/min/1.73 m(2); P = 0.90). After radiocontrast, urinary 15-isoprostane F2(t) levels in placebo patients increased significantly over baseline values (2.9+/-0.7 vs 10.3+/-2.1 ng/mg creatinine; P = 0.007), whereas urinary 15-isoprostane F2(t) levels in N acetylcysteine patients remained basically unchanged (3.5+/-0.5 vs 4.1+/-0.9 ng/mg creatinine; P = 0.63). Furthermore, N-acetylcysteine treatment led to lower levels of alpha-glutathione S-transferase than did placebo treatment (0.8+/-0.2 vs 2.4+/-0.7 micro g/g; P = 0.046). CONCLUSIONS: In chronic renal failure patients, the improvement in renal function induced by post-radiocontrast administration of N-acetylcysteine is strongly associated with suppression of oxidant stress-mediated proximal tubular injury. PMID- 15128883 TI - Activation of matrix metalloproteinase-2 causes peritoneal injury during peritoneal dialysis in rats. AB - BACKGROUND: Sclerosing peritonitis (SP) and encapsulating peritoneal sclerosis (EPS) are serious complications of continuous ambulatory peritoneal dialysis. Although we have shown previously that matrix metalloproteinase-2 (MMP-2) is increased in peritoneal injury leading to SP/EPS, most of the MMP-2 in the dialysate drained from the peritoneal cavity was the latent form that was lacking activity. In the present study, we investigated whether MMP-2 causes peritoneal injury. METHODS: To create an animal model of peritoneal injury, we administered intraperitoneally chlorhexidine gluconate to rats. Dialysate drained from these rats was analysed by gelatin zymography and MMP-2 activity was analysed by an in situ film zymography method. In vitro myofibroblasts were cultured in collagen three-dimensional culture and then MMP-2 in conditioned medium from the culture was analysed by gelatin zymography. RESULTS: Zymographic analysis revealed that latent form MMP-2 levels were high in the dialysate from peritoneal injury rats, whereas the active form was barely detectable. MMP-2 activity in the peritoneal tissue of the peritoneal injury rats was strongly detected by in situ film zymography. In vitro myofibroblasts were promoted to produce MMP-2 and to activate MMP-2 in collagen three-dimensional culture. CONCLUSIONS: In the present model, most of the MMP-2 was in the latent form, but activation of MMP-2 was promoted in the peritoneum during peritoneal injury. Activated MMP-2 may be associated with the progression of peritoneal injury. PMID- 15128884 TI - Serum osteoprotegerin levels and the extent of vascular calcification in haemodialysis patients. AB - BACKGROUND: Osteoprotegerin (OPG) is a glycoprotein that inhibits osteoclast differentiation and activity. OPG-deficient mice develop severe osteoporosis and medial arterial calcification. The expression of OPG is detected in early atherosclerotic lesions in non-uraemic patients. We examined whether serum OPG is associated with aortic calcification in haemodialysis patients. METHODS: Serum OPG was measured in 102 patients who were undergoing haemodialysis. The aortic calcification index (ACI) was assessed by computed tomography scans. RESULTS: Serum OPG level, measured by enzyme-linked immunosorbent assay, was significantly greater in patients with higher ACI than in those with lower ACI. There was a direct relationship between ACI and serum OPG levels and a positive association between OPG and ACI (r = 0.483, P<0.0001). Multiple regression analyses indicated that serum OPG levels were independently associated with the severity of aortic calcification (P<0.0001). CONCLUSIONS: These findings show that serum OPG levels are associated with the extent of vascular calcification, suggesting that OPG may be involved in the development of vascular calcification in haemodialysis patients. PMID- 15128885 TI - Not known from ADAM(TS-13)--novel insights into the pathophysiology of thrombotic microangiopathies. PMID- 15128886 TI - Medication prescribing patterns in ambulatory haemodialysis patients: comparisons of USRDS to a large not-for-profit dialysis provider. AB - BACKGROUND: End-stage renal disease (ESRD) patients are prescribed numerous medications. The United States Renal Data System (USRDS) reported on medication prescribing patterns in 1998. Since then, several new medications, treatment guidelines and recommendations have been introduced. The objective was to analyse and compare haemodialysis (HD) patient medication prescribing patterns between the Dialysis Clinic, Inc. (DCI) database and the USRDS report. METHODS: Point prevalent (01/01/03) medication use data from the DCI national database was obtained. Data collected included patient demographics, reason for and duration of ESRD, and medication listed on profile. All medications were classified similar to the USRDS and by where taken (clinic vs home). Medication prescribing patterns were compared between DCI and USRDS databases. Comparisons between age groups (<65 and >or=65 years) and diabetic status [diabetes mellitus (DM) vs non DM] were made. RESULTS: There were 128 477 medication orders categorized in 10 474 patients. DCI patient demographics were similar to present USRDS patients except for fewer Hispanics (P<0.001). Patients were prescribed 12.3+/-5.0 (median 12) different medications (2.6+/-1.4 clinic medications and 10.0+/-4.5 home medications). This is higher than reported by USRDS (median 9 medications). Patient age did not influence number of medications used (P = 0.54). DM patients are prescribed more medications than non-DM (13.3+/-5.0 DM vs 11.6+/-4.8 non-DM; P<0.00001). All medication class prescribing patterns were markedly different. CONCLUSION: The data suggest that medication prescribing patterns in HD patients have changed. The audit identified appropriate and questionable prescribing patterns. Various prescribing patterns identified areas for improvement in care (e.g. increased use of aspirin, beta-blockers and hyperlipidaemia medications) and areas requiring further investigation (e.g. high use of anti-acid, benzodiazepine and non-aluminum/non-calcium phosphate-binding medications). PMID- 15128887 TI - Histopathology of humorally mediated anti-glomerular basement membrane (GBM) glomerulonephritis in mice. AB - BACKGROUND: From a diagnostic point of view it would be important to learn more about the relationship between the immune responses underlying glomerulonephritis and the patterns of glomerular lesions. A murine model of anti-GBM glomerulonephritis in which inflammation is driven by delayed-type hypersensitivity (DTH) has been studied extensively. The aim of this study was to uncover histological features that might be specific for anti-GBM glomerulonephritis driven by a humoral immune response. METHODS: BALB/c mice were immunized with rabbit IgG in incomplete Freund's adjuvant. Six days later, on day 0, they received rabbit anti-GBM serum intravenously. Proteinuria was assessed with dipsticks. Mice were killed on days 4, 8 or 14. Kidneys from days 4 and 8 were processed for immunofluorescence and histology. On day 14 mice were perfusion-fixed for electron microscopy. RESULTS: Proteinuria started on day 3. Autologous IgG and of C3 were found along the GBM. There was only slight infiltration with macrophages and no measurable infiltration by CD4 T cells, indicating the virtual absence of DTH. Besides infiltration with neutrophils there were little histological alterations on day 4. On day 8 many loops were hyalinized. On day 14, cellular crescents were found in 23% of glomeruli. Subendothelial spaces contained hyaline material, cells and fibrin. Podocytes displayed effacement of foot processes and apical microprotrusions. Podocyte bridges were common. These alterations were identical to those reported in the standard model that produces a DTH-like inflammation. CONCLUSION: The qualitative pattern of histological damage in a murine model of anti-GBM glomerulonephritis does not depend on the underlying immunological process. PMID- 15128888 TI - The worsening of left ventricular hypertrophy is the strongest predictor of sudden cardiac death in haemodialysis patients: a 10 year survey. AB - BACKGROUND: Although the incidence of sudden cardiac death (SCD) is high among haemodialysis (HD) patients, there are few papers available on this topic. The aim of this study on a single-centre HD population observed over a 10 year period was to identify patient- and HD-related specific factors that might be associated with a higher risk of SCD. METHODS: The study included 123 patients (76 men; age 29-79 years) undergoing renal replacement therapy at our dialysis unit for at least 6 months. For each patient, routine laboratory tests were performed monthly, blood pressure was measured both at the start and the end of each dialysis session, haemoglobin and pre-dialysis serum K(+) were determined weekly, serum iPTH was assessed thrice yearly, and an echocardiographic study was performed annually to determine the left ventricular mass index (LVMi). The prevalence of cardiovascular (CV) co-morbidities, and the incidence of new events were also recorded. RESULTS: During the 10 years, 85 patients died-16 from SCD, 30 from cardiac causes (CC) other than SCD, and 39 from other causes (OC); 38 patients were still alive (AL) at the end of the observation period. Comparative analysis of SCD, CC, OC and AL, reveals that the male prevalence (13/3) was higher in SCD than in AL, while AL were younger than the deceased patients regardless of the cause of death (P<0.0001; ANOVA), the duration of arterial hypertension was higher in SCD (129+/-104 months; P = 0.0005; ANOVA), despite similar antihypertensive therapies, and the difference between LVMi at end-point and at inception (deltaLVMi) was significantly higher in SCD [+56+/-38 g/m(2) body surface area] compared with OC (-5+/-35), AL (-17+/-25) and even CC (7+/-30) (P<0.0001; ANOVA); finally, the prevalence of patients with ischaemic heart disease (IHD) was higher in the SCD group (11/5; P<0.0001, chi(2)). Univariate Cox regression analysis demonstrated that the factors increasing the risk of SCD were IHD (P = 0.002), the worsening of left ventricular hypertrophy (LVH) (P<0.0001), and the presence of long-lasting arterial hypertension (P = 0.001). An increase in LVH was the sole risk factor for SCD when comparing SCD with CC patients (P = 0.003). By multivariate Cox regression analysis deltaLVMi was identified as the strongest predictor of SCD (P<0.0001). CONCLUSION: While confirming the role of common CV risk factors for SCD in dialysis patients such as IHD and arterial hypertension, this study is the first to demonstrate that the worsening of LVH is the strongest predictor of sudden death. PMID- 15128889 TI - The battle for access--health care in Afghanistan. PMID- 15128890 TI - Understanding "diastolic" heart failure. PMID- 15128891 TI - Advances in the treatment of mucopolysaccharidosis type I. PMID- 15128892 TI - Hospitalists in the United States--mission accomplished or work in progress? PMID- 15128893 TI - Postoperative concurrent radiotherapy and chemotherapy for high-risk squamous cell carcinoma of the head and neck. AB - BACKGROUND: Despite the use of resection and postoperative radiotherapy, high risk squamous-cell carcinoma of the head and neck frequently recurs in the original tumor bed. We tested the hypothesis that concurrent postoperative administration of cisplatin and radiotherapy would improve the rate of local and regional control. METHODS: Between September 9, 1995, and April 28, 2000, 459 patients were enrolled. After undergoing total resection of all visible and palpable disease, 231 patients were randomly assigned to receive radiotherapy alone (60 to 66 Gy in 30 to 33 fractions over a period of 6 to 6.6 weeks) and 228 patients to receive the identical treatment plus concurrent cisplatin (100 mg per square meter of body-surface area intravenously on days 1, 22, and 43). RESULTS: After a median follow-up of 45.9 months, the rate of local and regional control was significantly higher in the combined-therapy group than in the group given radiotherapy alone (hazard ratio for local or regional recurrence, 0.61; 95 percent confidence interval, 0.41 to 0.91; P=0.01). The estimated two-year rate of local and regional control was 82 percent in the combined-therapy group, as compared with 72 percent in the radiotherapy group. Disease-free survival was significantly longer in the combined-therapy group than in the radiotherapy group (hazard ratio for disease or death, 0.78; 95 percent confidence interval, 0.61 to 0.99; P=0.04), but overall survival was not (hazard ratio for death, 0.84; 95 percent confidence interval, 0.65 to 1.09; P=0.19). The incidence of acute adverse effects of grade 3 or greater was 34 percent in the radiotherapy group and 77 percent in the combined-therapy group (P<0.001). Four patients who received combined therapy died as a direct result of the treatment. CONCLUSIONS: Among high-risk patients with resected head and neck cancer, concurrent postoperative chemotherapy and radiotherapy significantly improve the rates of local and regional control and disease-free survival. However, the combined treatment is associated with a substantial increase in adverse effects. PMID- 15128894 TI - Postoperative irradiation with or without concomitant chemotherapy for locally advanced head and neck cancer. AB - BACKGROUND: We compared concomitant cisplatin and irradiation with radiotherapy alone as adjuvant treatment for stage III or IV head and neck cancer. METHODS: After undergoing surgery with curative intent, 167 patients were randomly assigned to receive radiotherapy alone (66 Gy over a period of 6 1/2 weeks) and 167 to receive the same radiotherapy regimen combined with 100 mg of cisplatin per square meter of body-surface area on days 1, 22, and 43 of the radiotherapy regimen. RESULTS: After a median follow-up of 60 months, the rate of progression free survival was significantly higher in the combined-therapy group than in the group given radiotherapy alone (P=0.04 by the log-rank test; hazard ratio for disease progression, 0.75; 95 percent confidence interval, 0.56 to 0.99), with 5 year Kaplan-Meier estimates of progression-free survival of 47 percent and 36 percent, respectively. The overall survival rate was also significantly higher in the combined-therapy group than in the radiotherapy group (P=0.02 by the log-rank test; hazard ratio for death, 0.70; 95 percent confidence interval, 0.52 to 0.95), with five-year Kaplan-Meier estimates of overall survival of 53 percent and 40 percent, respectively. The cumulative incidence of local or regional relapses was significantly lower in the combined-therapy group (P=0.007). The estimated five-year cumulative incidence of local or regional relapses (considering death from other causes as a competing risk) was 31 percent after radiotherapy and 18 percent after combined therapy. Severe (grade 3 or higher) adverse effects were more frequent after combined therapy (41 percent) than after radiotherapy (21 percent, P=0.001); the types of severe mucosal adverse effects were similar in the two groups, as was the incidence of late adverse effects. CONCLUSIONS: Postoperative concurrent administration of high-dose cisplatin with radiotherapy is more efficacious than radiotherapy alone in patients with locally advanced head and neck cancer and does not cause an undue number of late complications. PMID- 15128895 TI - Diastolic heart failure--abnormalities in active relaxation and passive stiffness of the left ventricle. AB - BACKGROUND: Patients with signs and symptoms of heart failure and a normal left ventricular ejection fraction are said to have diastolic heart failure. It has traditionally been thought that the pathophysiological cause of heart failure in these patients is an abnormality in the diastolic properties of the left ventricle; however, this hypothesis remains largely unproven. METHODS: We prospectively identified 47 patients who met the diagnostic criteria for definite diastolic heart failure; all the patients had signs and symptoms of heart failure, a normal ejection fraction, and an increased left ventricular end diastolic pressure. Ten patients who had no evidence of cardiovascular disease served as controls. Left ventricular diastolic function was assessed by means of cardiac catheterization and echocardiography. RESULTS: The patients with diastolic heart failure had abnormal left ventricular relaxation and increased left ventricular chamber stiffness. The mean (+/-SD) time constant for the isovolumic-pressure decline (tau) was longer in the group with diastolic heart failure than in the control group (59+/-14 msec vs. 35+/-10 msec, P=0.01). The diastolic pressure-volume relation was shifted up and to the left in the patients with diastolic heart failure as compared with the controls. The corrected left ventricular passive-stiffness constant was significantly higher in the group with diastolic heart failure than in the control group (0.03+/-0.01 vs. 0.01+/-0.01, P<0.001). CONCLUSIONS: Patients with heart failure and a normal ejection fraction have significant abnormalities in active relaxation and passive stiffness. In these patients, the pathophysiological cause of elevated diastolic pressures and heart failure is abnormal diastolic function. PMID- 15128896 TI - Cord-blood transplants from unrelated donors in patients with Hurler's syndrome. AB - BACKGROUND: Hurler's syndrome (the most severe form of mucopolysaccharidosis type I) causes progressive deterioration of the central nervous system and death in childhood. Allogeneic bone marrow transplantation before the age of two years halts disease progression and prolongs life, but many children lack a bone marrow donor. We investigated the feasibility of using cord-blood transplants from unrelated donors and a myeloablative preparative regimen that did not involve total-body irradiation in young children with Hurler's syndrome. METHODS: Between December 1995 and October 2002, 20 consecutive children with Hurler's syndrome received busulfan, cyclophosphamide, and antithymocyte globulin before receiving cord-blood transplants from unrelated donors. The children were subsequently evaluated for engraftment, adverse effects, and effects on disease symptoms. RESULTS: Cord-blood donors had normal alpha-L-iduronidase activity (mean number of cells, 10.53x10(7) per kilogram of body weight) and were discordant for up to three of six HLA markers. Neutrophil engraftment occurred a median of 24 days after transplantation. Five patients had grade II or grade III acute graft-versus host disease; none had extensive chronic graft-versus-host disease. Seventeen of the 20 children were alive a median of 905 days after transplantation, with complete donor chimerism and normal peripheral-blood alpha-L-iduronidase activity (event-free survival rate, 85 percent). Transplantation improved neurocognitive performance and decreased somatic features of Hurler's syndrome. CONCLUSIONS: Cord blood from unrelated donors appears to be an excellent source of stem cells for transplantation in patients with Hurler's syndrome. Sustained engraftment can be achieved without total-body irradiation. Cord-blood transplantation favorably altered the natural history of Hurler's syndrome and thus may be important to consider in young children with this form of the disease. PMID- 15128897 TI - Clinical practice. Genital herpes. PMID- 15128898 TI - The pathogenesis of mycosis fungoides. PMID- 15128899 TI - Images in clinical medicine. Cor triatriatum. PMID- 15128901 TI - Management of cancer of the head and neck--a cocktail with your PORT? PMID- 15128902 TI - Eicosanoids in cystic fibrosis. PMID- 15128900 TI - Clinical problem-solving. Index of suspicion. PMID- 15128903 TI - Postmenopausal osteoporosis and strontium ranelate. PMID- 15128904 TI - Circulating angiogenic factors and preeclampsia. PMID- 15128905 TI - Risks of testosterone replacement. PMID- 15128906 TI - Parvovirus B19. PMID- 15128907 TI - Multidisciplinary management of lung cancer. PMID- 15128908 TI - A halt to neuroblastoma screening in Japan. PMID- 15128909 TI - Images in clinical medicine. Air embolism after the insertion of a central venous catheter. PMID- 15128910 TI - Hyperoxia decreases matrix metalloproteinase-9 and increases tissue inhibitor of matrix metalloproteinase-1 protein in the newborn rat lung: association with arrested alveolarization. AB - Matrix metalloproteinases (MMP) are likely effectors of normal lung development, especially branching morphogenesis, angiogenesis, and extracellular matrix degradation. Because hyperoxia exposure (>95% O(2)) from d 4 to 14 in newborn rat pups leads to arrest of alveolarization and mimics newborn chronic lung disease, we tested whether hyperoxia altered MMP-2 and -9 mRNA, protein, and enzymatic activity, and the mRNA and protein expression of the endogenous tissue inhibitor of MMP, TIMP-1. No changes due to hyperoxia exposure were observed in MMP-2 mRNA or pro-enzyme (72 kD) protein levels between d 6 and 14, although the overall protein mass and zymographic activity of the active (68 kD) enzyme were diminished (p < 0.05, ANOVA). However, hyperoxia significantly decreased levels of MMP-9 mRNA and pro-MMP-9 protein and diminished overall MMP-9 pro-enzyme activity. TIMP-1 mRNA was not elevated by hyperoxia until d 14, but protein levels were significantly (p < 0.001) elevated by hyperoxia from d 9 to 14. To estimate the potential of MMP inhibition to arrest alveolarization, administration of doxycycline (20 mg/kg, twice daily by gavage), a pan-MMP proteolysis inhibitor, arrested lung alveolarization. We conclude that hyperoxia decreases MMP-9 mRNA, protein, and activity and elevates TIMP-1 protein, and these changes have the potential to contribute to the arrest of normal lung development. PMID- 15128912 TI - Plasma 8-isoprostane is increased in preterm infants who develop bronchopulmonary dysplasia or periventricular leukomalacia. AB - Our aim was to assess the plasma free 8-epi-prostaglandin F(2alpha) (8 isoprostane) and ascorbyl radical as risk indicators for oxidative damage in extremely low birth weight infants (ELBWIs) and the effect of N-acetylcysteine (NAC) on these markers. Plasma samples were collected on days 3 and 7 of life from infants who were enrolled in a randomized, controlled trial in which i.v. NAC or placebo was administered to ELBWIs during the first week of life, with the aim of preventing bronchopulmonary dysplasia (BPD). Plasma 8-isoprostane was analyzed in 83 infants using an enzyme immunoassay kit. Ascorbyl radical concentration was measured in 61 infants with electron spin resonance spectroscopy. The 8-isoprostane concentrations were similar in the NAC and placebo groups. In infants who later developed BPD or died (n = 29), the median (range) 8-isoprostane concentration was significantly higher (p = 0.001) on day 3 and day 7 [50.0 pg/mL (19-360) and 57.0 pg/mL (14-460), respectively] than in survivors without BPD [n = 54; 34.5 pg/mL (5-240) and 39.5 pg/mL (7-400), respectively]. The 8-isoprostane levels increased significantly more (p < 0.05) in infants who later developed periventricular leukomalacia. NAC treatment or the later development of BPD was not related to the ascorbyl radical levels. The ascorbyl radical level decreased significantly in all groups from day 3 to day 7, but the difference between the groups was not significant. The mean (SD) ascorbyl radical level on day 3 was significantly higher (p < 0.01) in infants who later developed periventricular leukomalacia [287 (124) versus 194 (90)]. These data suggest that plasma 8-isoprostane could serve as a marker in assessing the risk for BPD development in ELBWIs. PMID- 15128911 TI - Biliary atresia is associated with CD4+ Th1 cell-mediated portal tract inflammation. AB - A proposed mechanism in the pathogenesis of biliary atresia involves an initial virus-induced, progressive T cell-mediated inflammatory obliteration of bile ducts. The aim of this study was to characterize the inflammatory environment present within the liver of infants with biliary atresia to gain insight into the role of a primary immune-mediated process versus a nonspecific secondary response to biliary obstruction. Frozen liver tissue obtained from patients with biliary atresia, neonatal giant cell hepatitis, total parenteral nutrition (TPN)-related cholestasis, choledochal cysts, and normal control subjects was used for fluorescent immunohistochemistry studies of cellular infiltrates, cytokine mRNA expression, and in situ hybridization for localization of cytokine-producing cells. Immunohistochemistry revealed increases in CD8(+) and CD4(+) T cells and Kupffer cells (CD68(+)) in the portal tracts of biliary atresia. Reverse transcription-PCR analysis of biliary atresia tissue showed a Th1-type cytokine profile with expression of IL-2, interferon-gamma, tumor necrosis factor-alpha, and IL-12. This profile was not seen in normal, neonatal hepatitis or choledochal cyst livers but was present in TPN-related cholestasis. In situ hybridization revealed that the Th1 cytokine-producing cells were located in the portal tracts in biliary atresia and in the parenchyma of TPN-related cholestasis. A distinctive portal tract inflammatory environment is present in biliary atresia, involving CD4(+) Th1 cell-mediated immunity. The absence of similar inflammation in other pediatric cholestatic conditions suggests that the portal tract inflammation in biliary atresia is not a secondary response to cholestasis but rather indicates a specific immune response involved in the pathogenesis of biliary atresia. PMID- 15128914 TI - Effects of antenatal betamethasone administration on fetal heart rate and behavior in twin pregnancy. AB - The efficacy of antenatal corticosteroid therapy in reducing the incidence of respiratory distress syndrome in twin pregnancy is controversial. The corticosteroid levels achieved with the current regimen might not be high enough to reach both siblings. In singleton fetuses, maternal betamethasone administration causes transient reductions in fetal heart rate (FHR), FHR variability, and body and breathing movements. We performed a prospective study to determine whether there are similar effects of betamethasone on FHR and behavior in preterm twin pregnancy and whether the effects occur similarly in both twin members. Eighteen women who were carrying twins received optimal corticosteroid treatment. Simultaneous recordings were made on twins before (day 0), during (days 1-2), and after (days 3-4) corticosteroid therapy using separate cardiotocography and ultrasound machines. Betamethasone administration was associated with significant transient decreases in basal FHR (day 1), FHR variability (days 2 and 3), and body and breathing movements (day 2). The overall changes in twins were similar to those previously found in singleton pregnancies. There was a high degree of association of response to betamethasone among individual members of twin pairs. The betamethasone-induced effects were unrelated to fetal sex, positioning, chorionicity, and discordance in size, but there was an effect of gestation on FHR. We conclude that the current regimen of antenatal corticosteroids used in preterm twin pregnancies results in observable physiologic and behavioral changes in twin pairs irrespective of their composition. This provides evidence that the achieved betamethasone levels are high enough to reach the compartment of either twin member. PMID- 15128913 TI - Use of air displacement plethysmography in the determination of percentage of fat mass in african american children. AB - The purpose of this study was to determine the ability of air displacement plethysmography (ADP) to estimate percentage of fat mass (%FM) in African American children. %FM was determined in 21 boys and 13 girls (11.0 +/- 1.4 y, 18.6 +/- 4.2 kg/m(2) [mean +/- SD]) by ADP (using six published densitometric equations) and dual-energy x-ray absorptiometry (DXA). Measures were done within 2 h of one another, in random order. Regardless of equation, %FM(ADP) was significantly correlated with %FM(DXA) (R(2) = 0.67-0.71, all p < 0.001). %FM(ADP) using the equation of Siri (%FM(ADP-Siri) 20.3 +/- 9.0) agreed most closely with %FM(DXA) (20.0 +/- 10.2, difference p = 0.729). Together, surface area artifact and bone mineral content per unit of bone-free fat-free mass accounted for 29% of the variance in the residual between methods. The correlation between %FM(ADP-Siri) and %FM(DXA) was not significant for those <35 kg (n = 10; R(2) = 0.084, p = 0.417). There was a trend toward %FM(ADP-Siri) underestimating %FM(DXA) in girls (-1.46 +/- 3.0%FM; p = 0.103) but not in boys (1.43 +/- 6.4%FM; p = 0.315). Predicted lung volume was 40.1% higher than measured lung volume (p < 0.001). %FM(ADP-Siri) determined using predicted lung volume was 23.5 +/- 8.9, higher than that using measured lung volume (p < 0.001) and higher than %FM(DXA) (p = 0.001). We conclude that in 9- to 14-y-old African American children and provided lung volume is measured, %FM using ADP with Siri's equation approximates that obtained by DXA. Body composition results determined by ADP in children <35 kg should be interpreted with caution. PMID- 15128915 TI - Peripheral neuropathy with ataxia in childhood as a result of the G8363A mutation in mitochondrial DNA. AB - Peripheral neuropathy has been identified in children with mitochondrial encephalomyopathies but not as a main clinical landmark. Here we report the clinical, electrophysiologic, biochemical, and genetic findings in a family who harbors the G8363A mutation in the tRNALys gene of mitochondrial DNA. Affected individuals presented with peripheral neuropathy and ataxia as the main clinical sign. Additional involvement included muscle weakness and multiple lipomatosis. Other common clinical characteristics associated with the G8363A mutation, such as cardiomyopathy and myoclonus epilepsy, were not observed. These findings suggest that a mitochondrial disease should be considered in the differential diagnosis of children with heredoataxic syndrome and peripheral neuropathy of unknown origin. PMID- 15128916 TI - Tumor necrosis factor-alpha allele lymphotoxin-alpha+250 is associated with the presence and severity of placental inflammation among preterm births. AB - Histologic inflammation of placenta has been associated with increased risk for bronchopulmonary dysplasia and periventricular leukomalacia among preterm infants. Tumor necrosis factor-alpha (TNF-alpha) plays a central role in the regulation of inflammation. Some alleles of TNF (LT-alpha+250, TNF-alpha-308, and TNF-alpha-238) have been associated with susceptibility and/or severity of many diseases characterized by inflammation and/or involving the immune system. To determine whether alleles of TNF-alpha affect the risk and/or the severity of chorioamnionitis, we examined the placentas of 101 preterm births (birth weight t) in either allele. These results raise the possibility that T2 deficient patients with mild mutations have been misinterpreted as normal by the coupled assay with tiglyl-CoA. PMID- 15128924 TI - Association of maternal histocompatibility at class II HLA loci with maternal microchimerism in the fetus. AB - For investigating the possible influence of maternal-fetal HLA compatibility on maternal microchimerism, DNA samples from blood of 120 maternal-fetal pairs were genotyped at two polymorphic loci: glutathione-S-transferase M1 (GSTM1) and angiotensin-converting enzyme (ACE). Informative pairs (mother heterozygous/fetus homozygous at one of the two loci) were then tested by quantitative real-time PCR for the noninherited maternal allele(s) and genotyped at the HLA-A, B, and C class I loci and/or at the DRB1 and/or DQB1 class II loci. Small numbers of maternal cells were detected in the circulation of 16 of the 30 informative second- and third-trimester fetuses. Comparison with HLA data suggested an association between microchimerism and maternal compatibility at the class II DRB1 and/or DQB1 HLA loci and with the maternal HLA-DQB1*0301 allele. There was no relationship between maternal microchimerism and maternal-fetal HLA compatibility at other HLA loci or with gestational age, fetal anomalies, or red cell or platelet isoimmunity. PMID- 15128925 TI - Biliary atresia and Th1 function: linking lymphocytes and bile ducts: commentary on the article by Mack et al. on page 79. PMID- 15128926 TI - Are artificial neural networks "ready to use" for decision making in the neonatal intensive care unit? Commentary on the article by Mueller et al. and page 11. PMID- 15128927 TI - Early alteration of structural and functional brain development in premature infants born with intrauterine growth restriction. AB - Placental insufficiency with fetal intrauterine growth restriction (IUGR) is an important cause of perinatal mortality and morbidity and is subsequently associated with significant neurodevelopmental impairment in cognitive function, attention capacity, and school performance. The underlying biologic cause for this association is unclear. Twenty-eight preterm infants (gestational age 32.5 +/- 1.9 wk) were studied by early and term magnetic resonance imaging (MRI). An advanced quantitative volumetric three-dimensional MRI technique was used to measure brain tissue volumes in 14 premature infants with placental insufficiency, defined by abnormal antenatal Doppler measurements and mean birth weights <10(th) percentile (1246 +/- 299 g) (IUGR) and in 14 preterm infants matched for gestational age with normal mean birth weights 1843 +/- 246 g (control). Functional outcome was measured at term in all infants by a specialized assessment scale of preterm infant behavior. Premature infants with IUGR had a significant reduction in intracranial volume (mean +/- SD: 253.7 +/- 29.9 versus 300.5 +/- 43.5 mL, p < 0.01) and in cerebral cortical gray matter (mean +/- SD: 77.2 +/- 16.3 versus 106.8 +/- 24.6 mL, p < 0.01) when measured within the first 2 wk of life compared with control premature infants. These findings persisted at term with intracranial volume (mean +/- SD: 429.3 +/- 47.9 versus 475.9 +/- 53.4 mL, p < 0.05) and cerebral cortical gray matter (mean +/- SD: 149.3 +/- 29.2 versus 189 +/- 34.2 mL, p < 0.01). Behavioral assessment at term showed a significantly less mature score in the subsystem of attention interaction availability in IUGR infants (p < 0.01). Cerebral cortical gray matter volume at term correlated with attention-interaction capacity measured at term (r = 0.45, p < 0.05). These results suggest that placental insufficiency with IUGR have specific structural and functional consequences on cerebral cortical brain development. These findings may provide insight into the structural-functional correlate for the developmental deficits associated with IUGR. PMID- 15128928 TI - Chronic maternal fluoxetine infusion in pregnant sheep: effects on the maternal and fetal hypothalamic-pituitary-adrenal axes. AB - Depression during pregnancy is frequently treated with the selective serotonin reuptake inhibitor, fluoxetine (FX). FX increases serotonergic neurotransmission and serotonin plays a role in the regulation of the hypothalamic-pituitary adrenal (HPA) axis. We have therefore investigated the effect of chronic administration of FX to the pregnant ewe on the maternal and fetal HPA axes. Nineteen late-gestation sheep were surgically prepared for chronic study of the fetus. FX (n = 7, 98.5 microg/kg/d) or sterile water (control, n = 8) was administered to the ewe for 8 d by constant rate i.v. infusion with an initial FX bolus dose of 70 mg. Maternal and fetal plasma ACTH and cortisol concentrations were determined at 0700 h each day. Maternal plasma ACTH concentrations fell on infusion d 2, but no changes were observed in maternal plasma cortisol concentrations. Fetal plasma ACTH concentrations increased on infusion d 7, and fetal plasma cortisol concentrations increased on infusion d 6, 7, and 8 in the FX group. In addition, the regression coefficient for the relationship between fetal ACTH and cortisol levels was significantly greater in the FX group compared with the control group. Thus, maternal FX treatment increased fetal plasma cortisol concentration. These results are of particular interest in the context that exposure of the fetus to excess glucocorticoids at critical windows during development has been shown to increase the risk of poor health outcomes in later life. PMID- 15128929 TI - Mechanical ventilation effect on surfactant content, function, and lung compliance in the newborn rat. AB - Studies of ventilator-associated lung injury in adult experimental animal models have documented that high tidal volume (TV) results in lung injury characterized by impaired compliance and dysfunctional surfactant. Yet, there is evidence that, in neonates, ventilation with a higher than physiologic TV leads to improved lung compliance. The purpose of our study was to evaluate how lung compliance and surfactant was altered by high TV ventilation in the neonate. We utilized a new model (mechanically air-ventilated newborn rats, 4-8 d old), and used 40 or 10 mL/kg TV strategies. Age-matched nonventilated animals served as controls. In all animals, dynamic compliance progressively increased after initiation of mechanical ventilation and was significantly greater than basal values after 60 min (p < 0.01). Lung lavage total surfactant with both TV strategies (p < 0.05) and the large aggregate fraction (only in TV = 40 mL/kg; p < 0.01) were significantly increased by 60 min of mechanical ventilation, compared with control animals. Ventilation with 40 mL/kg TV for 60 min adversely affected the lung surfactant surface-tension lowering properties (p < 0.01). After 180 min of ventilation with 40 mL/kg TV, the lung total surfactant content and dynamic compliance values were no longer distinct from the nonventilated animals' values. We conclude that, in the newborn rat, mechanical ventilation with a higher than physiologic TV increases alveolar surfactant content and, over time, alters its biophysical properties, thus promoting an initial but transient improvement in lung compliance. PMID- 15128930 TI - Comparison of short- and long-duration oxygen treatment after cerebral asphyxia in newborn piglets. AB - We tested whether reoxygenation with 100% O(2) for 5 min after experimental asphyxia in newborn piglets was as efficient as 100% O(2) for 20 min compared with room air. Forty-one anesthetized piglets, 1-3 d old, were randomized to cerebral hypoxemia-ischemia-hypercapnia (HIH) or control (n = 5). HIH was achieved by ventilation with 8% O(2), temporary occlusion of the common carotid arteries, and adding of CO(2). After 25 min, reoxygenation-reperfusion was started with 100% O(2) for 20 min (group 1, n = 12), 100% O(2) for 5 min (group 2, n = 12), or 21% O(2) (group 3, n = 12). All piglets were observed for 2 h. During reoxygenation-reperfusion, significantly higher blood pressure and more complete restoration of microcirculation (laser Doppler flow) in the cerebral cortex was found in both groups reoxygenated with 100% O(2) compared with 21% O(2) (regional cerebral blood flow >or=100% versus 70% of baseline, p = 0.04). Reoxygenation with 100% O(2) for 5 min was as efficient as 20 min. Oxygen delivery in cortex was significantly higher in groups 1 and 2 compared with group 3 (p = 0.03), but there were no significant differences in cerebral metabolic rate for oxygen. In the striatum, no significant differences in flow or extracellular glutamate, glycerol, and lactate/pyruvate ratio were found between the groups. In conclusion, after experimental asphyxia, newborn piglets can be reoxygenated as efficiently with 100% O(2) for only 5 min as 100% O(2) for 20 min compared with room air. PMID- 15128931 TI - Regression of retinopathy by squalamine in a mouse model. AB - The goal of this study was to determine whether an antiangiogenic agent, squalamine, given late during the evolution of oxygen-induced retinopathy (OIR) in the mouse, could improve retinal neovascularization. OIR was induced in neonatal C57BL6 mice and the neonates were treated s.c. with squalamine doses begun at various times after OIR induction. A system of retinal whole mounts and assessment of neovascular nuclei extending beyond the inner limiting membrane from animals reared under room air or OIR conditions and killed periodically from d 12 to 21 were used to assess retinopathy in squalamine-treated and untreated animals. OIR evolved after 75% oxygen exposure in neonatal mice with florid retinal neovascularization developing by d 14. Squalamine (single dose, 25 mg/kg s.c.) given on d 15 or 16, but not d 17, substantially improved retinal neovascularization in the mouse model of OIR. There was improvement seen in the degree of blood vessel tuft formation, blood vessel tortuosity, and central vasoconstriction with squalamine treatment at d 15 or 16. Single-dose squalamine at d 12 was effective at reducing subsequent development of retinal neovascularization at doses as low as 1 mg/kg. Squalamine is a very active inhibitor of OIR in mouse neonates at doses as low as 1 mg/kg given once. Further, squalamine given late in the course of OIR improves retinopathy by inducing regression of retinal neovessels and abrogating invasion of new vessels beyond the inner-limiting membrane of the retina. PMID- 15128932 TI - Biography of Mark A. Ratner. PMID- 15128933 TI - Genetic evidence that the human CYP2R1 enzyme is a key vitamin D 25-hydroxylase. AB - The synthesis of bioactive vitamin D requires hydroxylation at the 1 alpha and 25 positions by cytochrome P450 enzymes in the kidney and liver, respectively. The mitochondrial enzyme CYP27B1 catalyzes 1 alpha-hydroxylation in the kidney but the identity of the hepatic 25-hydroxylase has remained unclear for >30 years. We previously identified the microsomal CYP2R1 protein as a potential candidate for the liver vitamin D 25-hydroxylase based on the enzyme's biochemical properties, conservation, and expression pattern. Here, we report a molecular analysis of a patient with low circulating levels of 25-hydroxyvitamin D and classic symptoms of vitamin D deficiency. This individual was found to be homozygous for a transition mutation in exon 2 of the CYP2R1 gene on chromosome 11p15.2. The inherited mutation caused the substitution of a proline for an evolutionarily conserved leucine at amino acid 99 in the CYP2R1 protein and eliminated vitamin D 25-hydroxylase enzyme activity. These data identify CYP2R1 as a biologically relevant vitamin D 25-hydroxylase and reveal the molecular basis of a human genetic disease, selective 25-hydroxyvitamin D deficiency. PMID- 15128934 TI - HIV-1-infected monocyte-derived dendritic cells do not undergo maturation but can elicit IL-10 production and T cell regulation. AB - Dendritic cells (DCs) undergo maturation during virus infection and thereby become potent stimulators of cell-mediated immunity. HIV-1 replicates in immature DCs, but we now find that infection is not accompanied by many components of maturation in either infected cells or uninfected bystanders. The infected cultures do not develop potent stimulating activity for the mixed leukocyte reaction (MLR), and the DCs producing HIV-1 gag p24 do not express CD83 and DC lysosome-associated membrane protein maturation markers. If different maturation stimuli are applied to DCs infected with HIV-1, the infected cells selectively fail to mature. When DCs from HIV-1-infected patients are infected and cultured with autologous T cells, IL-10 was produced in 6 of 10 patients. These DC-T cell cocultures could suppress another immune response, the MLR. The regulation was partially IL-10-dependent and correlated in extent with the level of IL-10 produced. Suppressor cells only developed from infected patients, rather than healthy controls, and the DCs had to be exposed to live virus rather than HIV-1 gag peptides or protein. These results indicate that HIV-1-infected DCs have two previously unrecognized means to evade immune responses: maturation can be blocked reducing the efficacy of antigen presentation from infected cells, and T cell-dependent suppression can be induced. PMID- 15128935 TI - SecB is a bona fide generalized chaperone in Escherichia coli. AB - It is known that the DnaK and Trigger Factor (TF) chaperones cooperate in the folding of newly synthesized cytosolic proteins in Escherichia coli. We recently showed that despite a very narrow temperature range of growth and high levels of aggregated cytosolic proteins, E. coli can tolerate deletion of both chaperones, suggesting that other chaperones might be involved in this process. Here, we show that the secretion-dedicated chaperone SecB efficiently suppresses both the temperature sensitivity and the aggregation-prone phenotypes of a strain lacking both TF and DnaK. SecB suppression is independent of a productive interaction with the SecA subunit of the translocon. Furthermore, in vitro cross-linking experiments demonstrate that SecB can interact both co- and posttranslationally with short nascent chains of both secretory and cytosolic proteins. Finally, we show that such cotranslational substrate recognition by SecB is greatly suppressed in the presence of ribosome-bound TF, but not by DnaK. Taken together, our data demonstrate that SecB acts as a bona fide generalized chaperone. PMID- 15128936 TI - Enlarged meristems and delayed growth in plp mutants result from lack of CaaX prenyltransferases. AB - Meristems require a myriad of intercellular signaling pathways for coordination of cell division within and between functional zones and clonal cell layers. This control of cell division ensures a constant availability of stem cells throughout the life span of the meristem while limiting overproliferation of meristematic cells and maintaining the meristem structure. We have undertaken a genetic screen to identify additional components of meristem signaling pathways. We identified pluripetala (plp) mutants based on their dramatically larger meristems and increased floral organ number. PLURIPETALA encodes the alpha-subunit shared between protein farnesyltransferase and protein geranylgeranyltransferase-I. plp mutants also have altered abscisic acid responses and overall much slower growth rate. plp is epistatic to mutations in the beta-subunit of farnesyltransferase and shows a synergistic interaction with clavata3 mutants. plp mutants lead to insights into the mechanism of meristem homeostasis and provide a unique in vivo system for studying the functional role of prenylation in eukaryotes. PMID- 15128937 TI - Floral homeotic genes are targets of gibberellin signaling in flower development. AB - Gibberellins (GAs) are a class of plant hormones involved in the regulation of flower development in Arabidopsis. The GA-deficient ga1-3 mutant shows retarded growth of all floral organs, especially abortive stamen development that results in complete male sterility. Until now, it has not been clear how GA regulates the late-stage development of floral organs after the establishment of their identities within floral meristems. Various combinations of null mutations of DELLA proteins can gradually rescue floral defects in ga1-3. In particular, the synergistic effect of rga-t2 and rgl2-1 can substantially restore flower development in ga1-3. We find that the transcript levels of floral homeotic genes APETALA3 (AP3), PISTILLATA (PI), and AGAMOUS (AG) are immediately upregulated in young flowers of ga1-3 upon GA treatment. Using a steroid-inducible activation of RGA, we further demonstrated that these floral homeotic genes are transcriptionally repressed by RGA activity in young flowers whereas the expression of LEAFY (LFY) and APETALA1 (AP1) is not substantially affected. In addition, we observed the partial rescue of floral defects in ga1-3 by overexpression of AG. Our results indicate that GA promotes the expression of floral homeotic genes by antagonizing the effects of DELLA proteins, thereby allowing continued flower development. PMID- 15128938 TI - Formicine ants: An arthropod source for the pumiliotoxin alkaloids of dendrobatid poison frogs. AB - A remarkable diversity of bioactive lipophilic alkaloids is present in the skin of poison frogs and toads worldwide. Originally discovered in neotropical dendrobatid frogs, these alkaloids are now known from mantellid frogs of Madagascar, certain myobatrachid frogs of Australia, and certain bufonid toads of South America. Presumably serving as a passive chemical defense, these alkaloids appear to be sequestered from a variety of alkaloid-containing arthropods. The pumiliotoxins represent a major, widespread, group of alkaloids that are found in virtually all anurans that are chemically defended by the presence of lipophilic alkaloids. Identifying an arthropod source for these alkaloids has been a considerable challenge for chemical ecologists. However, an extensive collection of neotropical forest arthropods has now revealed a putative arthropod source of the pumiliotoxins. Here we report on the presence of pumiliotoxins in formicine ants of the genera Brachymyrmex and Paratrechina, as well as the presence of these ants in the stomach contents of the microsympatric pumiliotoxin-containing dendrobatid frog, Dendrobates pumilio. These pumiliotoxins are major alkaloids in D. pumilio, and Brachymyrmex and Paratrechina ants now represent the only known dietary sources of these toxic alkaloids. These findings further support the significance of ant-specialization and alkaloid sequestration in the evolution of bright warning coloration in poison frogs and toads. PMID- 15128939 TI - Role for glyoxalase I in Alzheimer's disease. AB - P301L mutant tau transgenic mice develop neurofibrillary tangles, a histopathologic hallmark of Alzheimer's disease and frontotemporal dementia (FTDP 17). To identify differentially expressed genes and to gain insight into pathogenic mechanisms, we performed a stringent analysis of the microarray dataset obtained with RNA from whole brains of P301L mutant mice and identified a single up-regulated gene, glyoxalase I. This enzyme plays a critical role in the detoxification of dicarbonyl compounds and thereby reduces the formation of advanced glycation end products. In situ hybridization analysis revealed expression of glyoxalase I in all brain areas analyzed, both in transgenic and control mice. However, levels of glyoxalase I protein were significantly elevated in P301L brains, as shown by Western blot analysis and immunohistochemistry. Moreover, a glyoxalase I-specific antiserum revealed many intensely stained flame shaped neurons in Alzheimer's disease brain compared with brains from nondemented controls. In addition, we examined a single nucleotide polymorphism predicting a nonconservative amino acid substitution at position 111 (E111A) in ethnically independent populations. We identified significant and consistent deviations from Hardy-Weinberg equilibrium, which points to the presence of selection forces. The E111A single nucleotide polymorphism was not associated with the risk for Alzheimer's disease in the overall population. Together, our data demonstrate the potential of transcriptomics applied to animal models of human diseases. They suggest a previously unidentified role for glyoxalase I in neurodegenerative disease. PMID- 15128940 TI - An invariant aspartic acid in the DNA glycosylase domain of DEMETER is necessary for transcriptional activation of the imprinted MEDEA gene. AB - Helix-hairpin-helix DNA glycosylases are typically small proteins that initiate repair of DNA by excising damaged or mispaired bases. An invariant aspartic acid in the active site is involved in catalyzing the excision reaction. Replacement of this critical residue with an asparagine severely reduces catalytic activity but preserves enzyme stability and structure. The Arabidopsis DEMETER (DME) gene encodes a large 1,729-aa polypeptide with a 200-aa DNA glycosylase domain. DME is expressed primarily in the central cell of the female gametophyte. DME activates maternal allele expression of the imprinted MEDEA (MEA) gene in the central cell and is required for seed viability. We mutated the invariant aspartic acid at position 1304 in DME to asparagine (D1304N) to determine whether the catalytic activity of the DNA glycosylase domain is required for DME function in vivo. Transgenes expressing wild-type DME in the central cell rescue seed abortion caused by a mutation in the endogenous DME gene and activate maternal MEA:GFP transcription. However, transgenes expressing the D1304N mutant DME do not rescue seed abortion or activate maternal MEA:GFP transcription. Whereas ectopic expression of the wild-type DME polypeptide in pollen is sufficient to activate ectopic paternal MEA and MEA:GUS expression, equivalent expression of the D1304N mutant DME in pollen failed to do so. These results show that the conserved aspartic acid residue is necessary for DME to function in vivo and suggest that an active DNA glycosylase domain, normally associated with DNA repair, promotes gene transcription that is essential for gene imprinting. PMID- 15128941 TI - Reduction in DNA-binding affinity of Cys2His2 zinc finger proteins by linker phosphorylation. AB - Cys(2)His(2) zinc finger proteins make up the largest class of transcription factors encoded in the genomes of higher eukaryotes. Recent studies of the Ikaros transcription factor demonstrated that this zinc finger protein undergoes cell cycle-dependent changes in association with DNA that seem to be due to phosphorylation of Thr or Ser residues in the linker regions connecting adjacent zinc finger domains. The high degree of conservation of this linker sequence within the Cys(2)His(2) superfamily suggested a common mechanism for the cell cycle-dependent modulation of DNA-binding affinity throughout this large class of transcription factors. The effects of linker phosphorylation on DNA-binding affinity were investigated through a direct comparison of the DNA-binding properties of four synthetic zinc finger proteins produced by native chemical ligation. The four proteins, comprising three zinc finger domains joined by two consensus Thr-Gly-Glu-Lys-Pro linkers, correspond to all four possible combinations of linker Thr phosphorylation states. Fluorescence-based DNA-binding studies of a specific DNA-binding site revealed that phosphorylation of a single linker reduced binding affinity approximately 40-fold, whereas phosphorylation of both linkers reduced binding affinity 130-fold. These results with purified components demonstrate that linker phosphorylation does, indeed, produce a significant reduction in DNA-binding affinity and support a model wherein a single cell cycle-dependent Ser/Thr kinase could simultaneously inactivate a large number of zinc finger transcription factors. PMID- 15128942 TI - Stone age bedding by the Sea of Galilee. PMID- 15128943 TI - Transgenic pigs expressing plant genes. PMID- 15128944 TI - Modifier screens in the mouse: time to move forward with reverse genetics. PMID- 15128945 TI - Translocation and cleavage of myocardial dystrophin as a common pathway to advanced heart failure: a scheme for the progression of cardiac dysfunction. AB - Advanced heart failure (HF) is the leading cause of death in developed countries. The mechanism underlying the progression of cardiac dysfunction needs to be clarified to establish approaches to prevention or treatment. Here, using TO-2 hamsters with hereditary dilated cardiomyopathy, we show age-dependent cleavage and translocation of myocardial dystrophin (Dys) from the sarcolemma (SL) to the myoplasm, increased SL permeability in situ, and a close relationship between the loss of Dys and hemodynamic indices. In addition, we observed a surprising correlation between the amount of Dys and the survival rate. Dys disruption is not an epiphenomenon but directly precedes progression to advanced HF, because long-lasting transfer of the missing delta-SG gene to degrading cardiomyocytes in vivo with biologically nontoxic recombinant adenoassociated virus (rAAV) vector ameliorated all of the pathological features and changed the disease prognosis. Furthermore, acute HF after isoproterenol toxicity and chronic HF after coronary ligation in rats both time-dependently cause Dys disruption in the degrading myocardium. Dys cleavage was also detected in human hearts from patients with dilated cardiomyopathy of unidentified etiology, supporting a scheme consisting of SL instability, Dys cleavage, and translocation of Dys from the SL to the myoplasm, irrespective of an acute or chronic disease course and a hereditary or acquired origin. Hereditary HF may be curable with gene therapy, once the responsible gene is identified and precisely corrected. PMID- 15128946 TI - Inhibition of primary human T cell proliferation by Helicobacter pylori vacuolating toxin (VacA) is independent of VacA effects on IL-2 secretion. AB - Recent evidence indicates that the secreted Helicobacter pylori vacuolating toxin (VacA) inhibits the activation of T cells. VacA blocks IL-2 secretion in transformed T cell lines by suppressing the activation of nuclear factor of activated T cells (NFAT). In this study, we investigated the effects of VacA on primary human CD4(+) T cells. VacA inhibited the proliferation of primary human T cells activated through the T cell receptor (TCR) and CD28. VacA-treated Jurkat T cells secreted markedly diminished levels of IL-2 compared with untreated cells, whereas VacA-treated primary human T cells continued to secrete high levels of IL 2. Further experiments indicated that the VacA-induced inhibition of primary human T cell proliferation was not attributable to VacA effects on NFAT activation or IL-2 secretion. We show here that VacA suppresses IL-2-induced cell cycle progression and proliferation of primary human T cells without affecting IL 2-dependent survival. Through the analysis of a panel of mutant VacA proteins, we demonstrate that VacA-mediated inhibition of T cell proliferation requires an intact N-terminal hydrophobic region necessary for the formation of anion selective membrane channels. Remarkably, we demonstrate that one of these mutant VacA proteins [VacA-Delta(6-27)] abrogates the immunosuppressive actions of wild type VacA in a dominant-negative fashion. We suggest that VacA may inhibit the clonal expansion of T cells that have already been activated by H. pylori antigens, thereby allowing H. pylori to evade the adaptive immune response and establish chronic infection. PMID- 15128947 TI - Reconstitution of retroviral fusion and uncoating in a cell-free system. AB - The molecular events underlying the immediate steps of retroviral uncoating, occurring after membrane fusion and leading to the formation of an active reverse transcription complex, are not known. To better understand these processes, we have developed a cell-free system that recapitulates these early steps of retroviral replication by using avian sarcoma and leukosis virus as a model retrovirus. The substrates used in this system are viral particles that are trapped before completing membrane fusion. These virions are induced to fuse out of endosomes and the viral cores are released into solution where they are amenable to biochemical manipulation. This system revealed that membrane fusion is not sufficient to stimulate the formation of a reverse transcription complex. Instead, ATP hydrolysis and cellular factors >5 kDa in size are required. Furthermore, later steps of avian sarcoma and leukosis virus reverse transcription were stimulated by nuclear factors. The cell-free system should now allow for the definition of retroviral uncoating mechanisms and facilitate the identification and characterization of the cellular factors involved. PMID- 15128948 TI - Neutrality, niches, and dispersal in a temperate forest understory. AB - A fundamental goal of ecology is to understand what controls the distribution and abundance of species. Both environmental niches and trade-offs among species in dispersal and competitive ability have traditionally been cited as determinants of plant community composition. More recently, neutral models have shown that communities of species with identical life-history characteristics and no adaptation to environmental niches can form spatial distribution patterns similar to those found in nature, so long as the species have a limited dispersal distance. If there is a strong correlation between geographic distance and change in environmental conditions, however, such spatial patterns can arise through either neutral or niche-based processes. To test these competing theories, we developed a sampling design that decoupled distance and environment in the understory plant communities of an old-growth, temperate forest. We found strong evidence of niche-structuring but almost no support for neutral predictions. Dispersal limitation acted in conjunction with environmental gradients to determine species' distributions, and both functional and phylogenetic constraints appear to contribute to the niche differentiation that structures community assembly. Our results indicate that testing a neutral hypothesis without accounting for environmental gradients will at best cause unexplained variation in plant distributions and may well provide misleading support for neutrality because of a correlation between geographic distance and environment. PMID- 15128949 TI - Rational design and characterization of a Rac GTPase-specific small molecule inhibitor. AB - The signaling pathways mediated by Rho family GTPases have been implicated in many aspects of cell biology. The specificity of the pathways is achieved in part by the selective interaction between Dbl family guanine nucleotide exchange factors (GEFs) and their Rho GTPase substrates. Here, we report a first generation small-molecule inhibitor of Rac GTPase targeting Rac activation by GEF. The chemical compound NSC23766 was identified by a structure-based virtual screening of compounds that fit into a surface groove of Rac1 known to be critical for GEF specification. In vitro it could effectively inhibit Rac1 binding and activation by the Rac-specific GEF Trio or Tiam1 in a dose-dependent manner without interfering with the closely related Cdc42 or RhoA binding or activation by their respective GEFs or with Rac1 interaction with BcrGAP or effector PAK1. In cells, it potently blocked serum or platelet-derived growth factor-induced Rac1 activation and lamellipodia formation without affecting the activity of endogenous Cdc42 or RhoA. Moreover, this compound reduced Trio or Tiam1 but not Vav, Lbc, Intersectin, or a constitutively active Rac1 mutant stimulated cell growth and suppressed Trio, Tiam1, or Ras-induced cell transformation. When applied to human prostate cancer PC-3 cells, it was able to inhibit the proliferation, anchorage-independent growth and invasion phenotypes that require the endogenous Rac1 activity. Thus, NSC23766 constitutes a Rac specific small-molecule inhibitor that could be useful to study the role of Rac in various cellular functions and to reverse tumor cell phenotypes associated with Rac deregulation. PMID- 15128950 TI - Temporal and regional differences in the olfactory proteome as a consequence of MeCP2 deficiency. AB - Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in the gene encoding MeCP2. By binding to methylated CpG dinucleotide promoter regions, MeCP2 acts as a transcriptional repressor, predicting that its absence might result in widespread aberrant gene transcription, leading to the RTT phenotype. Considering this potentially broad action of MeCP2 on expression and the complexity of the brain, especially during development, we approached the consequences of MeCP2 deficiency in a mouse model by using a temporal and regional proteomic strategy. We used the olfactory system (olfactory epithelium and bulb) because its attributes make it an excellent developmental model system. We find evidence of temporal and regional proteomic pattern differences between WT and MeCP2-deficient mice. It was possible to segregate these changes in protein expression into five biological function groups: cytoskeleton arrangement, chromatin modeling, energy metabolism, cell signaling, and neuroprotection. By combining the proteomic results with the RNA levels of the identified proteins, we show that protein expression changes are the consequence of differences in mRNA level or posttranslational modifications. We conclude that brain regions and ages must be carefully considered when investigating MeCP2 deficiency, and that not only transcription should be taken into account as a source for these changes, but posttranslational protein modifications as well. PMID- 15128951 TI - Uncoupling conformational change from GTP hydrolysis in a heterotrimeric G protein alpha-subunit. AB - Heterotrimeric G protein alpha (G alpha) subunits possess intrinsic GTPase activity that leads to functional deactivation with a rate constant of approximately 2 min(-1) at 30 degrees C. GTP hydrolysis causes conformational changes in three regions of G alpha, including Switch I and Switch II. Mutation of G202-->A in Switch II of G alpha(i1) accelerates the rates of both GTP hydrolysis and conformational change, which is measured by the loss of fluorescence from Trp-211 in Switch II. Mutation of K180-->P in Switch I increases the rate of conformational change but decreases the GTPase rate, which causes transient but substantial accumulation of a low-fluorescence G alpha(i1).GTP species. Isothermal titration calorimetric analysis of the binding of (G202A)G alpha(i1) and (K180P)G alpha(i1) to the GTPase-activating protein RGS4 indicates that the G202A mutation stabilizes the pretransition state-like conformation of G alpha(i1) that is mimicked by the complex of G alpha(i1) with GDP and magnesium fluoroaluminate, whereas the K180P mutation destabilizes this state. The crystal structures of (K180P)G alpha(i1) bound to a slowly hydrolyzable GTP analog, and the GDP.magnesium fluoroaluminate complex provide evidence that the Mg(2+) binding site is destabilized and that Switch I is torsionally restrained by the K180P mutation. The data are consistent with a catalytic mechanism for G alpha in which major conformational transitions in Switch I and Switch II are obligate events that precede the bond-breaking step in GTP hydrolysis. In (K180P)G alpha(i1), the two events are decoupled kinetically, whereas in the native protein they are concerted. PMID- 15128952 TI - Ethics in managed care. PMID- 15128954 TI - Real-world use of evidence-based treatments in community behavioral health care. AB - It is one thing to tout "best practices" and another thing to actually employ them. The author of this month's column provides us with a "fly on the wall" view of the experience of one community mental health center in implementing evidence based treatments for mental illnesses. If you have ever sat in a meeting at which you are supposed to be generating a best-practice approach and all you are hearing is a discussion of procedures rather than outcomes measurement, this column will be of interest. PMID- 15128955 TI - Atypical antipsychotic dosing: the effect of smoking and caffeine. PMID- 15128956 TI - Patients' early discontinuation of antidepressant prescriptions. PMID- 15128957 TI - Gifts from physicians to patients: an ethical dilemma. PMID- 15128958 TI - Population thinking as an adjunct to the clinical trial perspective. PMID- 15128959 TI - Supplementing supported employment with workplace skills training. AB - Introduction by the column editors: Supported employment, as designed for persons with serious and persistent mental illness, has been termed individual placement and support. In two randomized controlled trials (1,2), clients who received individual placement and support services were more likely to obtain at least one job in the competitive sector, to work more hours, and to have a higher total income than their counterparts who received more traditional types of vocational rehabilitation. However, individual placement and support did not improve the length of time the employed participants kept their jobs. An adjunctive or additional element of individual placement and support, aimed at improving the job tenure of individuals with mental illness, would be a constructive contribution to the vocational rehabilitation for this population. In a previous Rehab Rounds column, Wallace and colleagues (3) described the development of the workplace fundamental skills module, a highly structured and user-friendly curriculum designed to teach workers with mental illness the social and workplace skills needed to keep their jobs. The workplace fundamental skills module supplements individual placement and support by conveying specific skills that enable workers to learn the requirements of their jobs, anticipate the stressors associated with their jobs, and cope with stressors by using a problem-solving process. The earlier report described the production and validation of the module's content. The purpose of this month's column is to present the preliminary results of a randomized comparison of the module's effects on job retention, symptoms, and community functioning when coupled with individual placement and support. To enable wide generalization of the findings of the study, the program was conducted in a typical community mental health center. PMID- 15128960 TI - Current concepts in pharmacotherapy for posttraumatic stress disorder. AB - OBJECTIVE: This article describes current approaches to the pharmacologic treatment of posttraumatic stress disorder (PTSD) and reviews the classes of pharmacologic agents used in the treatment of PTSD. Pharmacotherapy for PTSD that is comorbid with other psychiatric disorders is highlighted. METHODS: The primary source literature was reviewed by using a MEDLINE search. Secondary-source review articles and chapters were also used. Results from studies of the psychophysiology of PTSD are outlined in the review to help inform treatment choices. The review gives more consideration to controlled studies than to open clinical trials. Recommendations for treatment are evidence based. RESULTS AND DISCUSSION: A growing body of evidence demonstrates the efficacy of pharmacologic treatment for PTSD. The effectiveness of the selective serotonin reuptake inhibitors sertraline and paroxetine in large-scale, well-designed, placebo controlled trials resulted in their being the first medications to receive approval from the U.S. Food and Drug Administration for the treatment of PTSD. Observation of psychophysiologic alterations associated with PTSD has led to the study of adrenergic-inhibiting agents and mood stabilizers as therapeutic agents. Controlled clinical trials with these classes of medication are needed to determine their efficacy for treating PTSD. Finally, the choice of medication for treating PTSD is often determined by the prominence of specific PTSD symptoms and the pattern of comorbid psychiatric conditions. PMID- 15128961 TI - The partners in care approach to ethics outcomes in quality improvement programs for depression. AB - OBJECTIVE: Patient centeredness and equity are major quality goals, but little is known about how these goals are affected by efforts to improve the quality of care. The authors describe an approach to addressing these goals in a randomized trial of quality improvement for depressed primary care patients. METHODS: For four ethics goals (autonomy, distributive justice, beneficence, and avoiding harm), the authors identify intervention features, study measures, and hypotheses implemented in Partners in Care, a randomized trial of two quality improvement interventions, relative to usual care and summarize published findings pertinent to these outcomes. RESULTS: To implement an ethics framework, modifications were required in study design and in measures and analysis plans, particularly to address the autonomy and justice goals. Extra resources were needed for sample recruitment, for intervention and survey materials, and to fund an ethics coinvestigator. The interventions were associated with improvements in all four ethics areas. Patients who received the interventions were significantly more likely to receive the treatment they had indicated at baseline as their preferred treatment (autonomy goal). Intervention-associated benefits occurred more rapidly among sicker patients and extended to patients from ethnic minority groups, resulting in a reduction in ethnic-group disparities in health outcomes relative to usual care (distributive justice goal). The interventions were associated with improved quality of care and health outcomes (beneficence goal) and with reduced use of long-term minor tranquilizers (goal of avoiding harm). CONCLUSION: S: It is feasible to explicitly address ethics outcomes in quality improvement programs for depression, but substantial marginal resources may be required. Nevertheless, interventions so modified can increase a practice's ability to realize ethics goals. PMID- 15128962 TI - An exploratory analysis of correlates of recovery. AB - OBJECTIVE: The concept of recovery has received increasing emphasis in the delivery of services to persons with schizophrenia. This study was an initial effort to develop an empirically based model of factors associated with a recovery orientation. METHODS: The authors reanalyzed data from 825 persons with schizophrenia who were assessed in the Schizophrenia Patient Outcomes Research Team (PORT) client survey. Multiple regression models were used to identify client and service use variables associated with each of four domains identified as important to a recovery orientation: life satisfaction, hope and optimism, knowledge about mental illness and services, and empowerment. RESULTS: In each regression model, the strongest relationship was observed between recovery orientation and lower severity of depressive symptoms. Both receipt of family psychoeducation and fewer side effects of medications were significantly and positively related to three of the four recovery domains. Psychotic symptoms were associated with less life satisfaction. Receipt of various services, including day treatment and legal services, was positively associated with knowledge about illness and services. CONCLUSIONS: Severity of psychiatric symptoms, a core feature of the biomedical perspective of mental illness, was negatively associated with a recovery orientation, and use of a variety of standard services were positively associated with a recovery orientation. Thus a polarized view of biomedical and recovery perspectives on mental illness may be unfounded, given that these perspectives appear to be mutually reinforcing. PMID- 15128963 TI - Treatment costs for youths receiving multisystemic therapy or hospitalization after a psychiatric crisis. AB - OBJECTIVE: The authors conducted a cost analysis for multisystemic therapy, an evidence-based treatment that is used as an intensive community-based alternative to the hospitalization of youths presenting with psychiatric emergencies. METHODS: Data from a randomized clinical trial that compared multisystemic therapy with usual inpatient services followed by community aftercare were used to compare Medicaid costs and clinical outcomes during a four-month period postreferral and a 12-month follow-up period. Data were from 115 families receiving Medicaid (out of 156 families in the clinical trial). RESULTS: During the four months postreferral, multisystemic therapy was associated with an average net savings per youth treated of $1,617 compared with usual services. Costs during the 12-month follow-up period were similar between treatments. Multisystemic therapy demonstrated better short-term cost-effectiveness for each of the clinical outcomes (externalizing behavior, internalizing behavior, and global severity of symptoms) than did usual inpatient care and community aftercare. The two treatments demonstrated equivalent long-term cost effectiveness. CONCLUSIONS: Among youths presenting with psychiatric emergencies, multisystemic therapy was associated with better outcomes at a lower cost during the initial postreferral period and with equivalent costs and outcomes during the 12-month follow-up period. PMID- 15128964 TI - Practitioner relationships and quality of care for low-income persons with serious mental illness. AB - OBJECTIVE: Though central to experiences of mental health care for persons with serious mental illness, relationships with practitioners have been underemphasized in recent quality-of-care research. This qualitative study described concepts of good care in relationships with psychiatrists, therapists, and case managers from the perspectives of low-income persons with psychiatric disabilities. METHODS: In-person, semistructured interviews were conducted with 51 adult Medicaid enrollees with psychiatric disabilities and diagnoses of schizophrenia. Grounded theory techniques were used to analyze the data. RESULTS: Eight categories representing service users' priorities for care in practitioner relationships resulted from the analysis: getting "extra things," looking for common ground, feeling known, the importance of talk, feeling like "somebody," practitioner availability, practitioner flexibility, and opportunities for input into treatment. CONCLUSIONS: Quantitative and illness-centered formulations may miss much of what low-income service users with serious mental illness value in their relationships with practitioners. The opportunity to counter feelings of vulnerability and alienation with a sense of connection that is based on shared humanness may be a high priority for services for this group. Practitioner relationships that help service users feel cared about and connected to the social world address suffering in mental illness and are thus essential to the meaning of good care. PMID- 15128965 TI - Use of claims data to examine the impact of length of inpatient psychiatric stay on readmission rate. AB - OBJECTIVE: This study analyzed the impact of length of stay for inpatient treatment of psychiatric disorders on readmission rates. METHODS: Hospitalization data were obtained from the MarketScan data set collected by Medstat. The instrumental variable method, an econometric technique, was used to estimate the impact of length of stay on the rate of readmission for 5,735 persons who had at least one discharge with a primary diagnosis of a psychiatric disorder during 1997 and 1998. RESULTS: Decreasing length of stay below ten days led to an increase in the readmission rate during the 30 days after discharge. Decreasing the length of stay from seven to six days increased the expected readmission rate from.04 to.047 (17.5 percent), whereas decreasing length of stay from four to three days increased the readmission rate from.09 to.136 (51.1 percent). CONCLUSION: Decreasing length of stay for inpatient psychiatric treatment increased the readmission rate. The use of instrumental variables could help better estimate the value of mental health services when using observational data. PMID- 15128966 TI - One-year housing arrangements among homeless adults with serious mental illness in the ACCESS program. AB - OBJECTIVE: This study examined the various living arrangements among formerly homeless adults with mental illness 12 months after they entered case management. METHODS: The study surveyed 5,325 clients who received intensive case management services in the Access to Community Care and Effective Services and Supports (ACCESS) program. Living arrangements 12 months after program entry were classified into six types on the basis of residential setting, the presence of others in the home, and stability (living in the same place for 60 days). Differences in perceived housing quality, unmet housing needs, and overall satisfaction were compared across living arrangements by using analysis of covariance. RESULTS: One year after entering case management, 37 percent of clients had been independently housed during the previous 60 days (29 percent lived alone in their own place and 8 percent lived with others in their own place), 52 percent had been dependently housed during the previous 60 days (11 percent lived in someone else's place, 10 percent lived in an institution, and 31 percent lived in multiple places), and 11 percent had literally been homeless during the previous 60 days. Clients with less severe mental health and addiction problems at baseline and those in communities that had higher social capital and more affordable housing were more likely to become independently housed, to show greater clinical improvement, and to have greater access to housing services. After the analysis adjusted for potentially confounding factors, independently housed clients were more satisfied with life overall. However, no significant association was found between specific living arrangements and either perceived housing quality or perceived unmet needs for housing. CONCLUSIONS: Living independently was positively associated with satisfaction of life overall, but it was not associated with the perception that the quality of housing was better or that there was less of a need for permanent housing. PMID- 15128967 TI - The impact of fear of HIPAA violation on patient care. AB - The deadline for compliance with the privacy rule of the Health Insurance Portability and Accountability Act of 1996 (HIPAA) was April 14, 2003. Understandably, many psychiatrists and other mental health providers have been focusing on compliance. This brief report discusses the negative impact of efforts to implement HIPAA on patient care in a community-based mental health system. Three cases highlight several issues: fear of violating HIPAA, failure to understand HIPAA's privacy regulations, and ethical concerns. The authors discuss the key issues and address implications for practice. Several recommendations are offered for maintaining excellent patient care while complying with HIPAA. PMID- 15128968 TI - Implications of educating the public on mental illness, violence, and stigma. AB - This study examined how two types of public education programs influenced how the public perceived persons with mental illness, their potential for violence, and the stigma of mental illness. A total of 161 participants were randomly assigned to one of three programs: one that aimed to combat stigma, one that highlighted the association between violence and psychiatric disorders, and a control group. Participants who completed the education-about-violence program were significantly more likely to report attitudes related to fear and dangerousness, to endorse services that coerced persons into treatment and treated them in segregated areas, to avoid persons with mental illness in social situations, and to be reluctant to help persons with mental illness. PMID- 15128969 TI - Reduction of episodes of seclusion and restraint in a psychiatric emergency service. AB - The authors developed a comprehensive plan focusing on the early identification and management of problematic behaviors in an effort to reduce seclusion and restraint in a psychiatric emergency service and to increase adherence to hospital standards for its use. Hospital data for nine months before and nine months after the implementation of the plan were retrospectively reviewed. Two key factors that were believed to increase the likelihood of episodes of seclusion and restraint were ineffectual management of problematic behavior and inadequate monitoring. The plan, when instituted, was associated with a 39 percent reduction of instances of seclusion and restraint. Compliance with hospital standards increased to 100 percent. PMID- 15128970 TI - Need for and use of mental health services among parents of children in the head start program. AB - This study examined the prevalence of psychosocial difficulties and use of mental health services among 290 parents of children in the Head Start program. Data on demographic characteristics, child behavior problems, parents' difficulties, home environment, child behavior, and use of health services were collected. A total of 161 parents (56 percent) had identifiable psychosocial difficulties, and 41 (14 percent) reported use of mental health services in the previous 12 months. Child behavior problems, unmet need for mental and physical health services, and less optimal home environments were associated with parents' psychosocial difficulties. Parents who had an unmet need for mental health services were more likely to report behavior problems among their children. PMID- 15128972 TI - Patients and families and DSM revision process. PMID- 15128974 TI - Training in transcultural psychiatry to reduce racial and ethnic disparities. PMID- 15128975 TI - Disability compensation for PTSD and use of VA mental health care. PMID- 15128977 TI - Vascular occlusion: can we push radiofrequency ablation into new size frontiers? PMID- 15128978 TI - Suspected appendicitis in children: in search of the single best diagnostic test. PMID- 15128979 TI - Scoring systems for CT in cystic fibrosis: who cares? PMID- 15128980 TI - The battle of intimal hyperplasia in the war against femoropopliteal disease. PMID- 15128981 TI - Coronary Artery MR Angiography: Are We There Yet? PMID- 15128982 TI - Metastatic renal cell carcinoma: CT-guided immunotherapy as a technically feasible and safe approach to delivery of gene therapy for treatment. AB - PURPOSE: To assess the technical feasibility and safety of weekly outpatient percutaneous computed tomographic (CT)-guided intratumoral injections of interleukin-2 (IL-2) plasmid DNA in a wide variety of superficial and deep tumor sites. MATERIALS AND METHODS: Twenty-nine patients with metastatic renal cell carcinoma and a total of 30 lesions measuring 1.0 cm(2) or greater in accessible thoracic (n = 15) or abdominal (n = 15) locations underwent up to three cycles of six weekly intratumoral IL-2 plasmid DNA injections. CT was used to guide needle placement and injection. After injection cycle 1, patients whose tumors demonstrated stable (< or =25% increase and < or =50% decrease in product of lesion diameters) or decreased size (>50% decrease in product of lesion diameters) advanced to injection cycle 2. Patients whose lesions decreased in size by more than 50% over the course of injection cycle 2 were eligible to begin injection cycle 3. An acceptable safety and technical feasibility profile for this technique was deemed to be (a) a safety and feasibility profile similar to that of single-needle biopsy and (b) an absence of serious adverse events (as defined in Title 21 of the Code of Federal Regulations) and/or unacceptable toxicities (as graded according to the National Cancer Institute Common Toxicity Criteria). RESULTS: A total of 284 intratumoral injections were performed, with a mean of 9.8 injections (range, 6-18 injections) received by each patient. Technical success (needle placement and injection of gene therapy agent) was achieved in all cases. Complications were experienced after 42 (14.8%) of the 284 injections. The most common complication was pneumothorax (at 32 [28.6%] of 112 intrathoracic injections), for which only one patient required catheter drainage. Complications occurred randomly throughout injection cycles and did not appear to increase as patients received more injections (P =.532). No patient experienced serious adverse events or unacceptable toxicities. CONCLUSION: Percutaneous CT guided intratumoral immunotherapy injections are technically feasible and can be safely performed. PMID- 15128983 TI - Evaluation of cystic renal masses: comparison of CT and MR imaging by using the Bosniak classification system. AB - PURPOSE: To compare computed tomography (CT) and magnetic resonance (MR) imaging in the evaluation of cystic renal masses by using the Bosniak classification system. MATERIALS AND METHODS: Images of 69 renal masses in 59 patients (38 men, 21 women; mean age, 60.4 years; range, 30-86 years), who had undergone both CT and MR imaging examinations within 1 year (average, 60.5 days; range, 0-356 days), were retrospectively analyzed by two radiologists in consensus. For each lesion, images were compared for thickness of wall and septa, number of septa, and presence of enhancement. Each mass was categorized (Bosniak classification) first on CT images and then on MR images, and results were compared. Pathologic correlation was available in 25 lesions. RESULTS: On CT images, there were 15 category I, 16 category II, 10 category IIF, 19 category III, and nine category IV lesions. Findings on CT and MR images were similar in 56 (81%) lesions; in 13 (19%) lesions, there were differences. In eight (12%) lesions, MR imaging depicted more septa than did CT, which resulted in an upgrade of the classification at MR imaging in two cases. In seven (10%) lesions, MR imaging depicted increased wall and/or septa thickness compared with CT, resulting in a classification upgrade in six cases. Three lesions had both increased numbers of septa and thickening of the wall and/or septa. In two (3%) lesions, enhancement characteristics at CT and MR imaging were different. One of these lesions also had an increased number of septa. Overall, MR imaging results led to a cyst classification upgrade of seven lesions, from category II to IIF (n = 2), IIF to III (n = 3), or III to IV (n = 2). Pathologic correlation in 25 lesions revealed 20 malignant and five benign lesions. CONCLUSION: CT and MR imaging findings were similar in the majority of cystic renal masses. In some cases, however, MR images may depict additional septa, thickening of the wall and/or septa, or enhancement, which may lead to an upgraded Bosniak cyst classification and can affect case management. PMID- 15128985 TI - Case 70: seminoma in an undescended testis. PMID- 15128986 TI - Gunshot wounds of abdomen: evaluation of stable patients with triple-contrast helical CT. AB - PURPOSE: To assess helical computed tomography (CT) with contrast material administered intravenously, orally, and rectally (triple contrast helical CT)) in the prospective evaluation of stable patients with abdominal gunshot wounds in whom there is no clinical indication for immediate exploratory laparotomy. MATERIALS AND METHODS: The study was conducted for 19 months. All patients met the following inclusion criteria: age of 16 years or older, hemodynamic stability, no clinical signs of peritoneal irritation, and signed consent to participate. Patients with obvious indications for laparotomy, such as gastrointestinal bleeding or evisceration, were excluded from the study. Forty seven patients fulfilled the criteria and underwent abdominal triple-contrast helical CT. CT findings were evaluated by one of four radiologists for evidence of peritoneal penetration and injury to solid organs or hollow viscera. Patients were followed up clinically for 13 weeks. CT findings were compared with those at surgery and/or clinical follow-up. RESULTS: CT demonstrated abnormalities in 27 (57%) patients. Laparotomy was performed in 11 (23%) patients; 10 procedures were therapeutic and one was nontherapeutic. The remaining 20 patients had a negative CT scan. These patients were treated conservatively. One injury was missed at CT. For prediction of the need for laparotomy, sensitivity of CT was 96%; specificity, 95%; positive predictive value, 96%; negative predictive value, 95%; and accuracy, 96%. CONCLUSION: In stable patients with gunshot wounds to the abdomen in whom there is no indication for immediate surgery, triple-contrast helical CT can help reduce the number of cases of unnecessary or nontherapeutic laparotomy (negative laparotomy) and can help identify patients with injuries that may be safely treated without surgery. PMID- 15128987 TI - Scirrhous gastric carcinoma: endoscopy versus upper gastrointestinal radiography. AB - PURPOSE: To compare the accuracy of upper gastrointestinal (UGI) series and endoscopic examination in the diagnosis and localization of scirrhous gastric carcinoma. MATERIALS AND METHODS: Seventy-two patients with pathologically proved scirrhous gastric carcinoma in surgical specimens were included. Preoperative reports at UGI series and endoscopic examination, which included impressions on the location and extent of the tumor, were compared with pathology reports, and the accuracy of the preoperative reports was calculated. Two gastrointestinal radiologists retrospectively reviewed the appearance of mucosa at UGI series. RESULTS: Preoperative diagnoses at endoscopy were Borrmann type IV carcinoma in 28 patients (39%), type III carcinoma in 29 (40%), early gastric carcinoma in seven (10%), lymphoma in six (8%), atrophic gastritis in one (3%), and type II carcinoma in one (3%). Preoperative diagnoses at UGI series were type IV carcinoma in 44 patients (61%), type III carcinoma in 25 (35%), lymphoma in two (3%), and early gastric carcinoma in one (1%). Pathology reports were compared with the preoperative reports, and tumor location and extent were correct in the endoscopic examination reports of 24 patients (33%) and the UGI series reports of 49 patients (68%). In 68 patients, UGI series revealed thickened and irregular folds in 62 (91%), ulceration in 42 (62%), and nodularity in 22 (32%) at consensus review. Endoscopic biopsy samples were positive for malignancy in 66 patients (93%). CONCLUSION: UGI series is superior to endoscopic examination in the diagnosis and localization of scirrhous gastric carcinoma. PMID- 15128988 TI - Radiation risks potentially associated with low-dose CT screening of adult smokers for lung cancer. AB - PURPOSE: To estimate the radiation-related lung cancer risks associated with annual low-dose computed tomographic (CT) lung screening in adult smokers and former smokers, and to establish a baseline risk that the potential benefits of such screening should exceed. MATERIALS AND METHODS: The estimated lung radiation dose from low-dose CT lung examinations corresponds to a dose range for which there is direct evidence of increased cancer risk in atomic bomb survivors. Estimated dose-, sex-, and smoking status-dependent excess relative risks of lung cancer were derived from cancer incidence data for atomic bomb survivors and used to calculate the excess lung cancer risks associated with a single CT lung examination at a given age in a U.S. population. From these, the overall radiation risks associated with annual CT lung screening were estimated. RESULTS: A 50-year-old female smoker who undergoes annual CT lung screening until age 75 would incur an estimated radiation-related lung cancer risk of 0.85%, in addition to her otherwise expected lung cancer risk of approximately 17%. The radiation associated cancer risk to other organs would be far lower. If 50% of all current and former smokers in the U.S. population aged 50-75 years received annual CT screening, the estimated number of lung cancers associated with radiation from screening would be approximately 36,000, a 1.8% (95% credibility interval: 0.5%, 5.5%) increase over the otherwise expected number. CONCLUSION: Given the estimated upper limit of a 5.5% increase in lung cancer risk attributable to annual CT-related radiation exposure, a mortality benefit of considerably more than 5% may be necessary to outweigh the potential radiation risks. PMID- 15128989 TI - Small pulmonary nodules: reproducibility of three-dimensional volumetric measurement and estimation of time to follow-up CT. AB - PURPOSE: To determine reproducibility of volume measurements of small pulmonary nodules on computed tomographic (CT) scans and to estimate critical time to follow-up CT. MATERIALS AND METHODS: One hundred fifteen pulmonary nodules for which two thin-section small-field-of-view CT scans were obtained and which were stable during 2-year observation were evaluated. A standard group of 94 nodules (with no or minimal artifact) and an expanded group of 105 nodules (including those with moderate artifacts) were examined. Percentage volume change (PVC) and monthly volumetric growth index (MVGI) were computed for each nodule pair. By using estimates of the variation in PVC in stable nodules as a function of initial diameter, critical time to follow-up CT was estimated; this time is the earliest point at which growth in a nodule of a given size can be reliably identified with repeat CT. RESULTS: The SD of PVC decreased with increasing nodule size from 18.5% in 2-5-mm nodules to 10.6% in 5-8-mm nodules and to 7.47% in 8-10-mm nodules. Inclusion of cases with moderate motion artifacts increased the SD of PVC to 27.4% in 2-5-mm nodules, to 17.1% in 5-8-mm nodules, and to 19.3% in 8-10-mm nodules. Critical time to follow-up CT for nodules detected at baseline screening was 12, 5, and 3 months and 1 month for those with initial sizes of 2, 5, 8, and 10 mm, respectively. For nodules detected at annual repeat screening, it was 4 and 3 months and 1 month for nodules that were 3, 4, and 5 mm or larger in size, respectively. Mean MVGI in 94 standard cases was 0.06%, and standard error was 0.21%. CONCLUSION: Factors that affect reproducibility of nodule volume measurements and critical time to follow-up CT include nodule size at detection, type of scan (baseline or annual repeat) on which the nodule is detected, and presence of patient-induced artifacts. PMID- 15128990 TI - Are two-dimensional CT measurements of small noncalcified pulmonary nodules reliable? AB - PURPOSE: To evaluate the intra- and interreader agreement of two-dimensional computed tomographic (CT) measurements of pulmonary nodules less than 2 cm in diameter. MATERIALS AND METHODS: Three readers independently made three serial measurements of each of 54 pulmonary nodules measuring 3-18 mm that had been observed on standard-dose multisection CT images obtained in 24 patients who ranged in age from 36 to 81 years (mean age, 54.6 years). There were 14 women (58%), who ranged in age from 43 to 81 years (mean age, 58.9 years), and 10 men (42%), who ranged in age from 36 to 65 years (mean age, 48.5 years). The largest transverse cross-sectional diameter of each nodule was measured at picture archiving and communication system, or PACS, workstations by using high-spatial resolution reconstructed CT images and identical window settings. Intra- and interreader agreement were determined by using methods described by Bland and Altman: the coefficient of repeatability for intrareader agreement, and methods derived from the 95% limits of agreement defined by Bland and Altman for interreader agreement. RESULTS: The repeatability coefficients were 1.70, 1.32, and 1.51 mm for readers 1, 2, and 3, respectively. The 95% limits of agreement for the difference among readers were -1.73 and 1.73. CONCLUSION: Two-dimensional CT measurements are not reliable in the evaluation of small noncalcified pulmonary nodules. PMID- 15128991 TI - Pulmonary nodules: preliminary experience with three-dimensional evaluation. AB - PURPOSE: To evaluate software designed to calculate pulmonary nodule volume in three dimensions. MATERIALS AND METHODS: Fifty-four solid noncalcified pulmonary nodules measuring 5-18 mm in diameter were studied with computed tomographic (CT) volumetric software. Baseline CT examinations were performed for various indications by using four-detector row multisection CT units, 1.25- or 2.50-mm sections, and a standard reconstruction algorithm. The percentage of successful nodule segmentations, as well as intraobserver variability, interreader agreement, and global repeatability of calculated volumes, was determined on the basis of consecutive measurements performed three times by three different radiologists by using the Bland and Altman method. The software was used to calculate the doubling time of 22 nodules for which a final diagnosis and comparable CT scans were available. RESULTS: Fifty-two (96%) of the 54 nodules were successfully segmented, allowing their volume to be calculated. Repeatability was high: There was no variation in the nine measurements of 35 (67%) of the 52 nodules. The coefficient of variation for the remaining 17 nodules (33%) was 2.26%. Bland and Altman 95% limits of acceptability, calculated on the basis of log-transformed data, yielded a maximum software measurement error of 6.38% of the previous volume measurement. Doubling time ranged from 4 to 188 years for the 13 benign nodules and from 37 to 216 days for the nine malignant nodules. CONCLUSION: Software volumetric analysis yielded repeatable estimates for 96% of the nodules examined. All software-calculated doubling times were in keeping with the benign or malignant nature of the nodules. PMID- 15128992 TI - Bronchiolitis obliterans syndrome in lung transplant recipients: can thin-section CT findings predict disease before its clinical appearance? AB - PURPOSE: To determine whether there are thin-section computed tomographic (CT) features that predict bronchiolitis obliterans syndrome (BOS) in lung transplant recipients before the clinical appearance and during the early stages of the disease. MATERIALS AND METHODS: Two hundred ninety-eight thin-section CT scans obtained in 26 lung transplant recipients who did (study group) and 26 lung transplant recipients who did not (control group) develop BOS were reviewed for the presence of mosaic perfusion, bronchiectasis, bronchial wall thickening, and air trapping. BOS was defined by using the recently revised definition of the International Society for Heart and Lung Transplantation. CT scans obtained in the BOS group were divided into three groups: Group A consisted of the last scans obtained before the clinical appearance of BOS; groups B and C consisted of, respectively, the first and last scans obtained after the clinical appearance of BOS. Scans obtained in the control group were acquired during similar posttransplantation periods and matched to scans in each BOS group. Sensitivity, specificity, and positive and negative predictive values were calculated separately for each subgroup. The optimal threshold for each thin-section CT depicted abnormality was defined by using receiver operating characteristics analysis. RESULTS: The sensitivities of air trapping for the diagnosis of BOS during the periods in which the scans in groups A, B, and C were obtained were 50%, 44%, and 64%, respectively; specificities were 80%, 100%, and 80% respectively. Sensitivities of mosaic perfusion were 4%, 20%, and 36%, respectively; specificities were 100%, 96%, and 96%, respectively. Sensitivities of bronchiectasis were 25%, 24%, and 32%, respectively; specificities were 80%, 80%, and 96%, respectively. Sensitivities of bronchial wall thickening were 4%, 24%, and 40%, respectively; specificities were 96%, 84%, and 80%, respectively. Air trapping was seen intermittently in nine (43%) of 21 patients with CT scans that depicted this finding at least once. CONCLUSION: The value of the finding of air trapping before the clinical appearance and during the early stages of BOS is lower than has been previously reported. When using the recently revised criteria for BOS, the role of thin-section CT as a screening test to evaluate patients with lung transplants appears to be limited. PMID- 15128993 TI - Hepatic perfusion changes in mice livers with developing colorectal cancer metastases. AB - PURPOSE: To evaluate whether intrahepatic flow alterations occur during formation of hepatic colorectal cancer metastases and to identify possible causes of these alterations. MATERIALS AND METHODS: Intravital imaging of exteriorized livers was performed in 72 live mice. Three groups of mice were studied: a sham-operated control group (n = 24), a group with nonmetastasizing subcutaneous gliomas (n = 24), and a group with developing hepatic CX-1 colon cancer metastases (n = 24). Microvascular flow parameters, leukocyte-endothelial interactions, and wall shear stress were directly measured in hepatic sinusoids and postsinusoidal venules at 2-day intervals prior to and during the development of metastases. The Kruskal Wallis test was used initially to test for overall equality of medians in each data group. Single posttest comparisons of independent samples were performed with the Mann-Whitney test, with an overall statistical significance of .05. RESULTS: Prior to the development of visible colorectal cancer metastases, significant (P <.05) reductions occurred in sinusoidal and postsinusoidal flow and wall shear rates, coupled with increased leukocyte rolling and adherence. With tumor growth, flow was further compromised in 92% of tumors larger than 0.5 mm in diameter by extrinsic compression of sinusoids and portal venules and narrowing caused by adherent leukocytes. CONCLUSION: Significant intrahepatic flow alterations occur in mouse livers prior to growth of visible metastases and provide a rational explanation for elevation in the Doppler perfusion index that occurs prior to tumor formation. PMID- 15128994 TI - Effects of perfusion on radiofrequency ablation in swine kidneys. AB - PURPOSE: To evaluate the effect of vascular occlusion on the size of radiofrequency (RF) ablation lesions and to evaluate embolization as an occlusion method. MATERIALS AND METHODS: The kidneys of six swine were surgically exposed. Fifteen RF ablation lesions were created in nine kidneys by using a 2-cm-tip single-needle ablation probe in varying conditions: Seven lesions were created with normal blood flow and eight were created with blood flow obstructed by means of vascular clamping (n = 5) or renal artery embolization (n = 3). The temperature, applied voltage, current, and impedance were recorded during RF ablation. Tissue-cooling curves acquired for 2 minutes immediately after the ablation were compared by using regression analysis. Lesions were bisected, and their maximum diameters were measured and compared by using analysis of variance. RESULTS: The mean diameter of ablation lesions created when blood flow was obstructed was 60% greater than that of lesions created when blood flow was normal (1.38 cm +/- 0.05 [standard error of mean] vs 0.86 cm +/- 0.07, P <.001). The two methods of flow obstruction yielded lesions of similar mean sizes: 1.40 cm +/- 0.06 with vascular clamping and 1.33 cm +/- 0.07 with embolization. The temperature at the probe tip when lesions were ablated with normal blood flow decreased more rapidly than did the temperature when lesions were ablated after flow obstruction (P <.001), but no significant differences in tissue-cooling curves between the two flow obstruction methods were observed. CONCLUSION: Obstruction of renal blood flow before and during RF ablation resulted in larger thermal lesions with potentially less variation in size compared with the lesions created with normal nonobstructed blood flow. Selective arterial embolization of the kidney vessels may be a useful adjunct to RF ablation of kidney tumors. PMID- 15128995 TI - Skeletal applications for flat-panel versus storage-phosphor radiography: effect of exposure on detection of low-contrast details. AB - PURPOSE: To compare exposure requirements for similar detection performance with flat-panel detectors and the most recent generation of storage-phosphor plates in the simulated scatter of typical skeletal radiographic examinations. MATERIALS AND METHODS: A contrast-detail test object was covered with varying thicknesses of acrylic to simulate skeletal exposure conditions in the wrist, knee, and pelvis. Three series were obtained with increasing thicknesses of a simulated soft-tissue layer (5, 10, and 20 cm) and increasing tube voltage (50, 70, and 90 kVp). A fourth series was obtained with exposure conditions adapted to the phantom instructions (75 kVp). Images were acquired with a flat-panel detector (cesium iodide scintillator) and storage-phosphor plates at five exposure levels (speed class range, 100-1,600). Five readers evaluated 84 images to determine the threshold contrast of 12 lesion diameters (range, 0.25-11.1 mm). Statistical significance of differences between the two digital systems was assessed with two way analysis of variance. RESULTS: A linear relationship was found between the number of detected lesions and the logarithm of exposure (R(2) > 0.98 for all series). On average, the flat-panel system required 45% less exposure than did the phosphor plates when 20-cm-thick acrylic was superimposed on the test object. Differences in exposure requirements were smaller with decreasing thicknesses of simulated soft-tissue layers and lower tube voltages (39% at 10 cm and 70 kVp, and 17% at 5 cm and 50 kVp). All differences were statistically significant. CONCLUSION: Flat-panel radiography provides improved contrast detectability and a potential for exposure reduction compared with those with storage-phosphor radiography. The best performance was achieved with conditions comparable to those for radiography of the trunk and lowest for conditions that simulate radiography of the extremities. PMID- 15128996 TI - The tram-track sign: cortical calcifications. PMID- 15128997 TI - Infrarenal aortic and lower-extremity arterial disease: diagnostic performance of multi-detector row CT angiography. AB - PURPOSE: To compare multi-detector row spiral computed tomographic (CT) angiography with digital subtraction angiography (DSA) in evaluation of the infrarenal aorta and lower-extremity arterial system. MATERIALS AND METHODS: Fifty patients with peripheral arterial occlusive disease were evaluated with multi-detector row CT angiography and DSA. Arteries depicted at CT angiography and DSA were graded separately for degree of stenosis as 23 anatomic segments (infrarenal aorta, right and left common iliac artery, internal iliac artery, external iliac artery, common femoral artery, superficial femoral artery, deep femoral artery, popliteal artery, anterior tibial artery, tibioperoneal trunk, posterior tibial artery, and peroneal artery). Grades included the following: 1, normal patency; 2, moderate (< or =50%) stenosis; 3, focal severe (>50%) stenosis; 4, multiple severe stenoses; and 5, occlusion. Three readers independently interpreted the images, and statistical analysis was performed. The results of image interpretation were evaluated for strength of agreement by using Cohen kappa statistics. On the basis of consensus readings, sensitivity, specificity, and accuracy for detection of stenotic lesions were calculated, with findings at DSA used as the reference standard. RESULTS: Substantial to almost perfect interobserver agreement was achieved in all cases. At DSA, 349 diseased segments were found among the 1,137 segments evaluated. Sensitivity, specificity, and accuracy, based on a consensus reading of multi-detector row CT angiograms, were 96%, 93%, and 94%, respectively. A statistically significant difference (P <.05) between DSA and multi-detector row CT angiography was present only in arteries graded 1 or 2. Interobserver agreement was almost perfect among the three readers for treatment recommendations based on findings at CT angiography and DSA. CONCLUSION: Multi-detector row CT angiography appears consistent and accurate in the assessment of patients with peripheral arterial occlusive disease. PMID- 15128998 TI - Electromagnetic breast imaging: average tissue property values in women with negative clinical findings. AB - Representative data are provided for three electromagnetic breast imaging techniques-near-infrared spectroscopy, electrical impedance spectroscopy, and microwave imaging spectroscopy-to serve as potential benchmarks for future investigation. The breasts of 23 women without clinical or mammographic findings of disease were imaged in the coronal plane with nonionizing radiation of varying frequencies. Average electromagnetic property values were reconstructed at each frequency on the basis of computational models of light diffusion, current flow, and microwave propagation. Electromagnetic properties were correlated with subject characteristics and between techniques. Each technique yielded information on breast tissue features (eg, conductivity, permittivity, light scattering, and absorption) that had not previously all been measured in the same individuals. PMID- 15128999 TI - MR imaging of newborns by using an MR-compatible incubator with integrated radiofrequency coils: initial experience. AB - To meet the needs of term and preterm neonates undergoing magnetic resonance (MR) imaging, an MR-compatible incubator with air, temperature, and humidity regulators and integrated radiofrequency coils was evaluated. Nine brain, two cardiac, and two pelvic examinations were performed by using a 1.5-T clinical MR imaging unit. The axillary temperature of the newborns varied by less than 0.8 degrees C, their vital signs remained stable, and no complications were encountered. The diagnostic quality of images obtained with the MR-compatible incubator was superior to that of images obtained with the standard MR imaging equipment. The use of an MR-compatible incubator for examinations of ill neonates is feasible and safe and yields excellent MR images. PMID- 15129000 TI - Acceptable rates of growth in fibroadenomas diagnosed with fine-needle aspiration biopsy. PMID- 15129001 TI - Endovascular abdominal aortic aneurysm repair: is delayed helical CT sufficient? PMID- 15129002 TI - Bland-Altman plot. PMID- 15129003 TI - Slacking off. DNA-break repair pathway poops out in older cells. PMID- 15129004 TI - If telomeres thrive, you will survive. Augmenting chromosome caps extends worm life span. PMID- 15129005 TI - Come one, come all. AB - Aging is a complex process that involves the gradual functional decline of many different tissues and cells. Gene expression microarray analysis provides a comprehensive view of the gene expression signature associated with age and is particularly valuable for understanding the molecular mechanisms that contribute to the aging process. However, because of the stochastic nature of the aging process, animals of the same chronological age often manifest great physiological differences. Therefore, profiling the gene expression pattern of a large population of aging animals risks either exaggerating or masking the changes in gene expression that correspond to physiological aging. In a recent paper, Golden and Melov surveyed the gene expression profiles of individual aging Caenorhabditis elegans, hoping to circumvent the problem of variability among worms of the same chronological age. This initial analysis of age-dependent gene expression in individual aging worms is an important step toward deciphering the molecular basis of physiological aging. PMID- 15129006 TI - Micromanagement of the ovarian follicle reserve--do stem cells play into the ledger? PMID- 15129007 TI - Endocrine mechanisms of intrauterine programming. AB - Epidemiological findings and experimental studies in animals have shown that individual tissues and whole organ systems can be programmed in utero during critical periods of development with adverse consequences for their function in later life. Detailed morphometric analyses of the data have shown that certain patterns of intrauterine growth, particularly growth retardation, can be related to specific postnatal outcomes. Since hormones regulate fetal growth and the development of individual fetal tissues, they have a central role in intrauterine programming. Hormones such as insulin, insulin-like growth factors, thyroxine and the glucocorticoids act as nutritional and maturational signals and adapt fetal development to prevailing intrauterine conditions, thereby maximizing the chances of survival both in utero and at birth. However, these adaptations may have long term sequelae. Of the hormones known to control fetal development, it is the glucocorticoids that are most likely to cause tissue programming in utero. They are growth inhibitory and affect the development of all the tissues and organ systems most at risk of postnatal pathophysiology when fetal growth is impaired. Their concentrations in utero are also elevated by all the nutritional and other challenges known to have programming effects. Glucocorticoids act at cellular and molecular levels to alter cell function by changing the expression of receptors, enzymes, ion channels and transporters. They also alter various growth factors, cytoarchitectural proteins, binding proteins and components of the intracellular signalling pathways. Glucocorticoids act, directly, on genes and, indirectly, through changes in the bioavailability of other hormones. These glucocorticoid induced endocrine changes may be transient or persist into postnatal life with consequences for tissue growth and development both before and after birth. In the long term, prenatal glucocorticoid exposure can permanently reset endocrine systems, such as the somatotrophic and hypothalamic-pituitary-adrenal axes, which, in turn, may contribute to the pathogenesis of adult disease. Endocrine changes may, therefore, be both the cause and the consequence of intrauterine programming. PMID- 15129008 TI - Concepts in sperm heterogeneity, sperm selection and sperm competition as biological foundations for laboratory tests of semen quality. AB - Stringent selection mechanisms, in both internal and external fertilisation systems, reject all but a significant minority of the spermatozoa released at ejaculation. Sperm competition theory provides circumstantial evidence that the selection process involves mechanisms by which the quality of the fertilising spermatozoon is controlled, thereby ensuring that females and their offspring receive high quality genetic material. In this review we examine some of these selection processes to see whether they could be exploited for the improvement of laboratory tests of sperm quality. Such tests are not only required for clinical and agricultural purposes, but are increasingly needed in fields such as reproductive and environmental toxicology where the species requirement is much broader. Despite many years of research, sperm quality assessment methods continue to provide imprecise data about fertility; here we suggest that this may be a consequence of using tests that focus on the spermatozoa that would normally be unable to fertilise under natural conditions. To achieve fertilisation a spermatozoon must be capable of responding appropriately to external signalling stimuli; those involving protein kinase-regulated flagellar function seem especially influential in governing effects ranging from non-Mendelian inheritance in mammals to sperm chemotaxis in sea urchins. Examination of the elicited responses reveals considerable heterogeneity in all species. Here we propose that this level of heterogeneity is meaningful both in terms of understanding how spermatozoa from some individuals possess fertility advantages over spermatozoa from their rivals in sperm competition, and in that the heterogeneity should be exploitable in the development of more accurate laboratory tests. PMID- 15129009 TI - Insulin increases 17 beta-estradiol production by the dominant follicle of the first postpartum follicle wave in dairy cows. AB - Prolonged anovulation following parturition has a negative impact on fertility in dairy cows. Insulin plays an important role in ovarian function in many species, and is profoundly depressed in dairy cows during early lactation. We hypothesized that hypoinsulinemia during early lactation represents a key indicator of nutritional status, resulting in delayed ovulation. Holstein cows (n = 10) were subjected to either a hyperinsulinemic-euglycemic clamp (INS) or saline infusion (CTL) for 96 h, beginning on day 10 after parturition during the first postpartum follicular wave. Insulin was infused continuously (0.3 microg/kg body weight per h) via a jugular catheter, and euglycemia was maintained by infusion of glucose. Circulating insulin concentrations were elevated 2.6-fold in INS cows compared with CTL cows (0.73 +/- 0.026 vs 0.28 +/- 0.026 ng/ml; P < 0.001). Insulin treatment did not affect (P > 0.05) luteinizing hormone (LH) pulse frequency, pulse amplitude or mean circulating LH. Circulating estradiol was elevated in INS cows (P < 0.01) and circulating testosterone also tended to be higher. The ratio of testosterone to estradiol was not different between treatments for the initial 30 h of infusion, but was significantly reduced thereafter in response to insulin (P < 0.01), suggesting that hyperinsulinemia increased follicular aromatase activity. Insulin treatment also resulted in reduced circulating nonesterified fatty acids, and increased circulating total and free insulin-like growth factor I concentrations. Insulin infusion increased estradiol secretion by the dominant follicle of the first postpartum follicular wave in dairy cows, and this effect appears not to be mediated through changes in pulsatile LH release. PMID- 15129010 TI - Acrosomal integrity and capacitation are not influenced by sperm cryopreservation in the giant panda. AB - Sperm cryopreservation and artificial insemination are important management tools for giant panda breeding and the preservation of extant genetic diversity. This study examined the influence of freeze-thawing on sperm function, specifically capacitation. Sperm from nine giant pandas were assessed before and after rapid ( 40 and - 100 degrees C/min) cryopreservation by incubation in HEPES-buffered Ham's F10 medium with and without the capacitation accelerators, 3-isobutyl-1 methylxanthine (IBMX) and dibutyryl cyclic AMP (dbcAMP). At 0, 3 and 6 h of exposure, aliquots were assessed for sperm motility traits and capacitation, defined as the proportion of sperm with intact acrosomes following exposure to solubilised zonae pellucidae (ursid or felid) or calcium ionophore subtracted from the proportion of sperm with intact acrosomes before exposure. Although mean+/-S.E.M. sperm motility post-thaw (56.1 +/- 3.9% at 0 h) was less (P < 0.05) than pre-freeze (71.7 +/- 6.0%), there was no difference (P > 0.05) in the proportion of acrosome-intact sperm (fresh, 93.0 +/- 1.7% versus cryopreserved thawed, 81.7 +/- 4.7% at 0 h). Incidence of capacitation was greater (P < 0.05) in fresh sperm incubated with capacitation accelerators IBMX and dbcAMP (9 h: 50.9 +/- 1.1) compared with fresh sperm incubated without accelerators (9 h: 41.2 +/- 1.1%). Frozen-thawed sperm preincubated without accelerators underwent capacitation (49.6 +/- 1.1%) to a greater extent (P < 0.05) compared with these fresh counterparts. Thawed samples with (9 h: 45.9 +/- 1.4%) and without accelerators (9 h: 41.2 +/- 1.1%) did not differ (P > 0.05) during the 9-h incubation. We conclude that giant panda spermatozoa (1) undergo capacitation in vitro with or without chemical accelerators and (2) withstand a rapid cryopreservation protocol, including retaining normal acrosomal integrity and functional capacitation ability. PMID- 15129011 TI - Birth of lambs of a pre-determined sex after in vitro production of embryos using frozen-thawed sex-sorted and re-frozen-thawed ram spermatozoa. AB - The characteristics and functional capacity of ram spermatozoa frozen-thawed prior to and after flow cytometric sorting was assessed after incubation (37 degrees C; 6 h), in vitro fertilisation (IVF), and transfer of fresh and vitrified in vitro produced embryos. Frozen-thawed spermatozoa from two rams were allocated to four treatment groups: (i) non-sorted (CONTROL); (ii) sorted (FS); (iii) sorted then re-frozen (FSF) and (iv) re-frozen control (FCF). Frozen-thawed samples were separated into X- and Y-chromosome bearing spermatozoa using a high speed sperm sorter after density gradient centrifugation (X: 88 +/- 1.5% and Y: 87 +/- 1.1% purity). After 6 h incubation (37 degrees C), the percentage of motile spermatozoa was higher (P < 0.001) for FS (84 +/- 2.0%) compared with all other treatments ( CONTROL: 36 +/- 3.3%, FSF: 28 +/- 3.1%, FCF: 20 +/- 2.0%). In a sperm migration test greater numbers of FS spermatozoa penetrated 5 mm into the artificial cervical mucus compared with spermatozoa from all other treatments (152 +/- 39.4 vs 31 +/- 9.2 spermatozoa respectively; P < 0.05). Fertilisation and cleavage rates were higher (P < 0.05) for in vitro matured oocytes inseminated with CONTROL compared with FSF spermatozoa. However, the Day 7 blastocyst development rate was higher for oocytes inseminated with FSF (62.2%) than FS and CONTROL spermatozoa (52.7 and 50.0%; P < 0.05). The number of ewes pregnant (Day 60), lambing and the in vivo embryo survival rate was greater (P < 0.01) after the transfer of fresh embryos rather than vitrified embryos derived from X- and Y-spermatozoa (67.6, 64.7 and 41.2% vs 29.6, 25.9 and 14.8% respectively). Twenty-six of the 30 (86.7%) lambs derived from sex-sorted spermatozoa were of the correct sex. These results demonstrate that frozen-thawed ram spermatozoa can be sex-sorted for immediate or future use after re cryopreservation and, in conjunction with IVF and embryo transfer, can be used to efficiently produce offspring of pre-determined sex. PMID- 15129012 TI - Methods for quantifying follicular numbers within the mouse ovary. AB - Accurate estimation of the number of ovarian follicles at various stages of development is an important indicator of the process of folliculogenesis in relation to the endocrine signals and paracrine/autocrine mechanisms that control the growth and maturation of the oocytes and their supporting follicular cells. There are 10-fold or greater differences in follicular numbers per ovary at similar ages and/or strains reported in earlier studies using various methods, leading to difficulties with interpretation of ovarian function in control vs experimental conditions. This study describes unbiased, assumption-free stereological methods for quantification of early and growing follicular numbers in the mouse ovary. A fractionator approach was used to sample a defined fraction of histological sections of adult wild-type ovaries. Primordial and primary follicles were counted independently with the optical and physical disector methods. The fractionator/disector methods, which are independent of follicular size or shape, gave estimations of 1930 +/- 286 (S.E.M.) and 2227 +/- 101 primordial follicles, and 137 +/- 25 and 265 +/- 32 primary follicles per ovary at 70 and 100 days of age respectively. From exact counts on serial sections, secondary and later follicular numbers at 100 days of age were estimated at 135 per ovary. Remnants of zona pellucidae (a marker of previous follicular atresia) were estimated using a fractionator/physical disector approach and were approximately 500 per ovary. The application of the quantitative methods described will facilitate an improved understanding of follicular dynamics and the factors that mediate their growth and maturation and allow for a better comparison between different studies. PMID- 15129013 TI - Boar seminal immunosuppressive fraction attenuates the leptin concentration and restores the thymus mass during pregnancy in mice. AB - The immunosuppressive fraction (ISF) of boar seminal vesicle fluid was recently demonstrated to inhibit production of T helper (Th)1 cytokines and enhance production of Th2 cytokines. The present study shows the effect of the ISF on leptin concentrations in blood plasma and adipose tissue in mice during pregnancy. The ISF effect on thymus weight during pregnancy is also demonstrated. The leptin concentration in blood plasma and adipose tissue increased and remained high in the latter half of pregnancy. ISF treatment at the beginning of pregnancy significantly lowered the leptin concentration both in blood plasma and adipose tissue of pregnant mice. Thymus involution has been described previously in pregnant mice. ISF treatment compensated for the loss of thymus mass during the whole pregnancy in the ISF-treated mice. The treatment of pregnant mice with ISF did not affect pregnancy and litter size. PMID- 15129014 TI - Components of litter size in mice after 110 generations of selection. AB - The aim of the present study was to evaluate how ovulation rate and survival rate through pregnancy had been affected by more than 110 generations of upwards selection on litter size in mice. The mean number of pups born alive was 22 in the high line (selected line) and 11 in the control line (an increase in 2.6 standard deviations). Selection on litter size increased ovulation rate by 4.6 standard deviations, and it is suggested that selection also increased embryonic mortality in late pregnancy. Embryo survival from ovulation until birth was 66% in the selected line and 69% in the control line, and the observed loss in litter size from day 16 of pregnancy until birth was possibly higher in the high line compared with the control line. Selection for higher litter size has significantly increased body weight in both males and females, as the mean weight at mating for the females was 46 g in the high line and 33 g in the control line respectively. PMID- 15129015 TI - Equine seminal plasma reduces sperm binding to polymorphonuclear neutrophils (PMNs) and improves the fertility of fresh semen inseminated into inflamed uteri. AB - Seminal plasma (SP) is known to have immunosuppressive properties in several species. Equine SP has been reported to reduce or inhibit chemotaxis, phagocytosis and complement activity in vitro. The type and amount of the SP component that suppresses sperm-polymorphonuclear neutrophil (PMN) binding in vitro was determined, and the effect of such suppression on the fertility of mares inseminated in the presence of uterine inflammation, was analyzed. Sperm cells were suspended in either SP, semen extender or a mixture of both, and each was mixed with PMN-rich uterine secretions collected at 12 h after artificial insemination (AI). SP reduced binding between spermatozoa and PMNs significantly (P < 0.05). Fertile spermatozoa were suspended in SP or semen extender and used to inseminate mares 12 h after the induction of uterine inflammation. The pregnancy rate was normal (77%) when spermatozoa were suspended in SP, but was dramatically reduced to only 5% when spermatozoa were suspended in extender. The proteins from SP, blood plasma (BP) and a skim-milk-based semen extender (skim milk extender, SME) were precipitated by ammonium sulfate, resuspended in PBS and dialyzed. The effect of the precipitated proteins on sperm-PMN binding was compared with fresh, untreated SP. Both fresh SP, and isolated SP proteins reduced sperm-PMN binding (P < 0.001). Conversely, proteins isolated from either BP or SME did not reduce sperm-PMN binding. The different concentrations of SP proteins used showed a dose-dependent suppression of sperm-PMN binding. Concentrations of 1 mg/ml SP protein significantly reduced sperm-PMN binding and 6 mg/ml reduced the binding to a level similar to that observed with fresh whole SP (P < 0.001). Finally, SP protein digested with proteinase K resulted in the complete loss of SP suppressive activity confirming that the effective component is a proteinaceous substance. PMID- 15129016 TI - Angiotensin II stimulates cAMP production and protein tyrosine phosphorylation in mouse spermatozoa. AB - Angiotensin II (AII), found in seminal plasma, has been shown to stimulate capacitation in uncapacitated mammalian spermatozoa. The present study investigated the location of AII receptors on spermatozoa and AII's mechanism of action. AT1 type receptors for AII are present on the acrosomal cap region and along the whole of the flagellum of both mouse and human spermatozoa. Because combinations of low concentrations of AII and either calcitonin or fertilization promoting peptide (FPP), both known to regulate the adenylyl cyclase (AC)/cAMP signal transduction pathway, elicited a significant response, this study investigated the hypothesis that these peptides act on the same pathway. AII was shown to significantly stimulate cAMP production in both uncapacitated and capacitated mouse spermatozoa and this was associated with increases in protein tyrosine phosphorylation. Using an anti-phosphotyrosine antibody to visualize the location of tyrosine phosphoproteins within individual cells, AII significantly stimulated phosphorylation within 20 min in both the head, especially in the acrosomal cap region, and the flagellum, especially in the principal piece, of uncapacitated mouse spermatozoa; combined AII + FPP was stimulatory within 5 min. In addition, Western blotting revealed that AII stimulation increased phosphorylation in a number of tyrosine phosphoproteins in both uncapacitated and capacitated mouse spermatozoa, with some being altered only in the latter category of cells. These results support the hypothesis that AII stimulates AC/cAMP in mammalian spermatozoa. PMID- 15129017 TI - Intravascular neutrophils partially mediate the endometrial endothelial cell proliferative response to oestrogen in ovariectomised mice. AB - The aim of this study was to investigate the role of intravascular neutrophils in initiating endothelial cell proliferation following oestrogen treatment in ovariectomised mouse endometrium. Uterine tissues were collected from ovariectomised C57/CBA female mice 24 h after oestrogen treatment with or without systemic neutrophil depletion. Neutropenia was achieved with either an in-house anti-neutrophil serum (ANS) or Gr-1 monoclonal antibody. All mice received an i.p. injection of bromodeoxyuridine (BrdU) 4 h prior to dissection to allow visualisation of proliferating cells using immunocytochemistry. Endometrial sections were immunostained for BrdU, vascular endothelial growth factor (VEGF), and neutrophils (using ANS). Oestrogen treatment of ovariectomised mice significantly increased the number of intravascular neutrophils, whereas induction of neutropenia with either ANS or Gr-1 in conjunction with oestrogen treatment prevented this increase. Oestrogen treatment of ovariectomised mice also significantly increased the number of intravascular VEGF-positive cells; however, whereas induction of neutropenia with ANS significantly reduced this increase, Gr-1 did not. In both studies, neutropenia significantly reduced, but did not eliminate, the amount of endometrial endothelial cell proliferation. These results suggest a role for neutrophils in endometrial angiogenesis following acute oestrogen treatment; however, the presence of VEGF-positive cells even after induction of neutropenia suggests that more than one type of leukocyte may be involved. PMID- 15129018 TI - Association between ovarian follicle development and pregnancy rates in dairy cows undergoing spontaneous oestrous cycles. AB - Ovarian follicle development continues in a wave-like manner during the bovine oestrous cycle giving rise to variation in the duration of ovulatory follicle development. The objectives of the present study were to determine whether a relationship exists between the duration of ovulatory follicle development and pregnancy rates following artificial insemination (AI) in dairy cows undergoing spontaneous oestrous cycles, and to identify factors influencing follicle turnover and pregnancy rate and the relationship between these two variables. Follicle development was monitored by daily transrectal ultrasonography from 10 days after oestrus until the subsequent oestrus in 158 lactating dairy cows. The cows were artificially inseminated following the second observed oestrus and pregnancy was diagnosed 35 days later. The predominant pattern of follicle development was two follicle waves (74.7%) with three follicle waves in 22.1% of oestrous cycles and four or more follicle waves in 3.2% of oestrous cycles. The interval from ovulatory follicle emergence to oestrus (EOI) was 3 days longer (P < 0.0001) in cows with two follicle waves than in those with three waves. Ovulatory follicles from two-wave oestrous cycles grew more slowly but were approximately 2 mm larger (P < 0.0001) on the day of oestrus. Twin ovulations were observed in 14.2% of oestrous cycles and occurred more frequently (P < 0.001) in three-wave oestrous cycles; consequently EOI was shorter in cows with twin ovulations. Overall, 57.0% of the cows were diagnosed pregnant 35 days after AI. Linear logistic regression analysis revealed an inverse relationship between EOI and the proportion of cows diagnosed pregnant, among all cows (n = 158; P < 0.01) and amongst those with single ovulations (n = 145; P < 0.05). Mean EOI was approximately 1 day shorter (P < 0.01) in cows that became pregnant than in non pregnant cows; however, pregnancy rates did not differ significantly among cows with different patterns of follicle development. These findings confirm and extend previous observations in pharmacologically manipulated cattle and show, for the first time, that in dairy cows undergoing spontaneous oestrous cycles, natural variation in the duration of post-emergence ovulatory follicle development has a significant effect on pregnancy rate, presumably reflecting variation in oocyte developmental competence. PMID- 15129019 TI - Respiratory response to toluene diisocyanate depends on prior frequency and concentration of dermal sensitization in mice. AB - Occupational asthma is the principal cause of work-related respiratory disease in the industrial world. In the absence of satisfactory models for predicting the potential of low molecular weight chemicals to cause asthma, we verified that dermal sensitization prior to intranasal challenge influences the respiratory response using toluene diisocyanate (TDI), a known respiratory sensitizer. BALB/c mice received TDI or vehicle (acetone/olive oil) on each ear on three consecutive days (days 1, 2, and 3; 0.3 or 3% TDI) or only once (day 1, 1% TDI). On day 7, the mice received similar dermal applications of vehicle or the same concentration of TDI as before ("boost"). On day 10, they received an intranasal dose of TDI (0.1%) or vehicle. Ventilatory function was monitored by whole body plethysmography for 40 min after intranasal application, and reactivity to inhaled methacholine was assessed 24 h later. Pulmonary inflammation was assessed by bronchoalveolar lavage and histology. Mice that received an intranasal dose of TDI without having received a prior dermal application of TDI did not exhibit any ventilatory response or inflammatory changes compared to vehicle controls. In contrast, mice that had received prior application(s) of TDI, even if only on day 7, exhibited the following: ventilatory responses, compatible with bronchoconstriction, immediately after intranasal application with TDI; enhanced methacholine responsiveness 24 h later; and pulmonary inflammation characterized by neutrophils. This was, however, not the case in mice that received the highest dermal amount of TDI (3% on days 1, 2, and 3). These findings suggest that respiratory response to TDI depends on prior frequency and concentration of dermal sensitization in mice. PMID- 15129020 TI - Species and tissue differences in the toxicity of 3-butene-1,2-diol in male Sprague-Dawley rats and B6C3F1 mice. AB - 3-Butene-1,2-diol (BDD) is a major metabolite of 1,3-butadiene (BD), but the role of BDD in BD toxicity and carcinogenicity remains unclear. In this study, the acute toxicity of BDD was investigated in male Sprague-Dawley rats and B6C3F1 mice. Of the rats given 250 mg/kg BDD, 2 out of 4 died within 24 h; rats experienced hypoglycemia, significant alterations of liver integrity tests, and had lesions in the liver 4 h after treatment, but no lesions were detected in extrahepatic tissues. Rat hepatic GSH and GSSG levels were significantly depleted at both 1 and 4 h after the BDD treatment. Rats administered 200 mg/kg BDD also had liver lesions but no death or hypoglycemia was observed four or 24 h after treatment; these rats had depleted hepatic GSH and GSSG levels at 1 h but not at 4 or 24 h after treatment. Mice administered 250 mg/kg BDD exhibited modest alterations of liver integrity tests, but no death, hypoglycemia, or lesions in any tissue, and hepatic GSH and GSSG levels were depleted at 1 h but not at 4 h. The plasma half-life of BDD was four times longer in rats than in mice. Additional studies in rats showed the depletion of hepatic GSH and GSSG preceded the BDD-induced hypoglycemia and hepatotoxicity. Thus, the long half-life of BDD in rat plasma and the sustained depletion of hepatic GSH and GSSG may in part explain the higher sensitivity of the rat to BDD-induced hepatotoxicity. Furthermore, the results indicate that BDD may play a role in BD-induced toxicity. PMID- 15129021 TI - Application of a statistical dynamic model investigating the short-term cellular kinetics induced by riddelliine, a hepatic endothelial carcinogen. AB - In recent studies, riddelliine, a pyrrolizidine alkaloid, was found to increase rates of replication and apoptosis and induce hemangiosarcoma in the liver of rats and mice. To analyze DNA replication and apoptosis data taken from the same animals, we have developed a predictive mathematical model for describing BrdU labeling and apoptotic processes. The model allows the incorporation of simple diurnal patterns in cellular kinetics and is applied to data on hepatocytes and endothelial cells taken from riddelliine exposed rats. Predictions from the model were used with multivariable nonlinear regression techniques to estimate replication and apoptotic rate constants for both cell types and all treatment groups. Hypothesis tests were used with the predicted rates to separate the competing effects of riddelliine on replication and apoptosis of hepatocytes and endothelial cells as well as compare replication rates between cell types. That estimated replication rates were found to be significantly higher for endothelial cells supports the supposition of induction of hemangiosarcoma by riddelliine in the liver. PMID- 15129023 TI - Human carcinogenic risk evaluation: an alternative approach to the two-year rodent bioassay. AB - An approach to the evaluation of carcinogenic risk resulting from exposure to a given chemical is presented in place of a reliance on two-year rodent bioassays. An emphasis is placed on evaluation of the potential DNA reactivity or increased cell proliferation that can be produced by a chemical. The special cases of immunosuppressive and estrogenic chemicals are considered. These evaluations are proposed to involve a combination of in vitro assays, computerized models, and short-term (up to 13 weeks) bioassays in rodents. The emphasis is on mechanistic understanding and evaluation of the dose response and relevance to humans. PMID- 15129022 TI - Expression of metallothionein isoform 3 (MT-3) determines the choice between apoptotic or necrotic cell death in Cd+2-exposed human proximal tubule cells. AB - This laboratory has shown that the third isoform of metallothionein (MT-3) is expressed in the human kidney in situ, including the cells of the proximal tubule. A subsequent analysis of MT-3 expression in cell cultures derived from the human proximal tubule (HPT) demonstrated that mortal HPT cells expressed MT 3, while the HPV-immortalized HK-2 cells had no expression of MT-3. In the present study, the effect of MT-3 expression on Cd(+2)-induced cytotoxicity was determined by stable transfection of the MT-3 coding sequence into the HK-2 cell line. The results demonstrated that HK-2 cells stably transfected with MT-3 were more sensitive to the cytotoxic effects of Cd(+2). Furthermore, this increase in Cd(+2)-induced cytotoxicity was correlated to an alteration in the mechanism of cell death, being changed from an apoptotic mechanism in cells not expressing the MT-3 gene to a necrotic mechanism in cells expressing the MT-3 gene. The present study provides evidence that MT-3 could play a role in controlling the choice between apoptosis and necrosis in multiple epithelial cell types of the human kidney. PMID- 15129024 TI - Induction of ERK1/2 and histone H3 phosphorylation within the outer stripe of the outer medulla of the Eker rat by 2,3,5-tris-(glutathion-S-yl)hydroquinone. AB - 2,3,5-tris-(glutathion-S-yl)-hydroquinone (TGHQ), a metabolite of hydroquinone (HQ), generates reactive oxygen species (ROS) in cultured renal epithelial cells and binds to tissue macromolecules within the rat kidney. The potential mechanisms by which TGHQ induces nephrotoxicity and nephrocarcinogenesis have been examined in cell culture models, but less is known concerning the molecular mechanisms of TGHQ-induced nephrotoxicity in vivo. In LLC-PK1 cells, TGHQ induces phosphorylation of both mitogen-activated protein kinase and histone H3, which likely promotes inappropriate chromatin condensation and mitotic catastrophe. Using the Eker (Tsc-2 mutant) rat as a model, we show by immunohistochemistry that TGHQ (7.5 micromol/kg) selectively induces ERK1/2 phosphorylation within the outer stripe of the outer medulla (OSOM) of the kidney. ERK1/2 phosphorylation is time-dependant, occurring as early as 1 h following treatment, and reaching maximal levels by 4 h. Subsequently, ERK1/2 phosphorylation returns to baseline levels by 24 h post treatment. ERK1/2 phosphorylation was confirmed by western blot analysis of OSOM tissue. Increases in histone H3 phosphorylation occurred subsequent to ERK1/2 phosphorylation (8 h), and reached a peak by 24 h, coincident with histological evidence of tissue necrosis. In contrast to studies in cell culture, neither JNK/SAPK nor p38 MAPK phosphorylation were significantly altered after TGHQ administration in vivo, as evidenced by western blot and immunohistochemical analyses. These data indicate that activation of the ERK1/2 pathway precedes overt cytotoxicity and that the signaling pathways activated by TGHQ in vivo and in vitro differ. PMID- 15129025 TI - [Clinical results of lateral closing wedge high tibial osteotomy for osteoarthritis of the knee (Oblique osteotomy with tension band fixation)]. AB - OBJECTIVES: We evaluated the preliminary results of oblique and lateral closing wedge high tibial osteotomy for medial compartment osteoarthritis of the knee. METHODS: Thirty-nine patients (29 women, 10 men; mean age 53 years; range 34 to 64 years) underwent oblique and lateral closing-wedge high tibial osteotomy followed by tension band plate fixation. According to the Ahlback system, seven patients had grade II, 27 patients had grade III, and five patients had grade IV osteoarthritis. Fixation was completed with a blade plate and two cortical screws. The results were evaluated using the Knee Society Score at the end of a mean follow-up of 23 months (range 12 to 41 months). RESULTS: The mean pre- and postoperative Knee Society scores were 43 (range 18-72) and 80 (range 20-90), and the mean Knee Function scores were 57 (range 45-90) and 72 (range 35-90), respectively (p<0.05). The mean preoperative deviation from the mechanical axis of the leg was 8.9 degrees varus (range 3 to 15 degrees). A mean correction of 11.6 degrees valgus (range 7 to 18 degrees) was afforded in order to obtain a slight valgus alignment. The mean postoperative femorotibial angle was 171 degrees (range 162-183 degrees). Complications were seen in 11 patients, which included severe overcorrection, fixation failure, transient nerve palsy, or pain over the fibular osteotomy site. CONCLUSION: Oblique high tibial osteotomy combined with tension band fixation is an effective procedure providing secure and durable fixation to allow early motion. It should be recalled that a high complication rate is likely during the learning curve, which adversely influences the clinical results. PMID- 15129026 TI - [The effectiveness of arthroscopic debridement and lavage treatment in osteoarthritis of the hip: preliminary results]. AB - OBJECTIVES: We evaluated the early results of arthroscopic debridement and lavage procedure in osteoarthritis of the hip. METHODS: The hips of 13 patients (7 women, 6 men; mean age 52 years; range 29 to 74 years) were treated by arthroscopic debridement and lavage in lateral position under general anesthesia. Ten patients had primary coxarthrosis; it was due to acetabular dysplasia in two patients, and to avascular necrosis of the femoral head in one patient. The patients were radiographically and arthroscopically evaluated according to the Kellgren-Lawrence grading system and the Tippett's chondropathy classification, respectively. Preoperative and postoperative assessments of the hips were made according to the Harris hip scoring system. The mean follow-up was 11.2 months (range 5 to 22 months). RESULTS: Harris hip scores increased in eight patients (62%) postoperatively, namely, in only one patient out of six with a radiographically high grade of osteoarthritis (grade 4), and in all seven patients with grade 2 or 3 disease. No complications occurred. Significant negative correlations were found between radiographic and arthroscopic grades and the clinical improvement (p=0.001). However, the patients' age and the preoperative Harris hip score were not correlated with the clinical result. CONCLUSION: Arthroscopic debridement and lavage provides significant clinical improvement during short-term follow-up of osteoarthritic hips with radiographic grades of 3 or less. PMID- 15129027 TI - [A comparison between locked intramedullary nailing and plate-screw fixation in the treatment of tibial diaphysis fractures]. AB - OBJECTIVES: The results of treatment with plate-screw or locked intramedullary nailing were evaluated for tibial diaphysis fractures, with a comparison of time to union and complications. METHODS: The study included 44 patients (31 males, 13 females; mean age 38 years; range 14 to 64 years) with tibial diaphysis fractures. The fractures were on the left side in 23 patients and on the right side in 20 patients. One patient had bilateral involvement. Twenty-six fractures were treated by intramedullary nailing and 19 fractures by plate-screw fixation after a mean time of nine days and 10 days from injury, respectively. The patients were evaluated with regard to operation time, range of motion, time to union, and complications after a mean follow-up of 23 months (range 18 to 46 months). RESULTS: The mean operation time and the mean range of motion of the ankle and knee were found similar in both groups. The mean time to union was five months with intramedullary nailing, and 3.5 months with plate-screw fixation. Length discrepancy occurred in only two patients (2 and 2.5 cm) in whom intramedullary nailing was performed. One patient treated by intramedullary nailing for a type III open fracture developed osteomyelitis. Plain radiographs showed angulation in four patients and in three patients following intramedullary nailing and plate-screw fixation, respectively, all of whom had distal-third tibial fractures. CONCLUSION: Plate-screw osteosynthesis yields satisfactory results in uncomminuted fractures of the tibial diaphysis, whereas locked intramedullary nailing is more appropriate in comminuted fractures because of better preservation of periosteal circulation, and thus lower complication rates. PMID- 15129028 TI - [Long-term results of open surgical repair of rotator cuff tears]. AB - OBJECTIVES: We evaluated the long-term results of rotator cuff tears treated by open surgical repair. METHODS: Ninety shoulders of 88 patients (36 females, 52 males; mean age 57 years; range 36 to 75 years) with rotator cuff tears were treated by open surgical repair and acromioplasty. A three-staged rehabilitation program was implemented following surgery. Objective evaluations were made with the use of the Constant-Murley scoring system. Pain was assessed through a visual analog scale. The mean follow-up period was 61 months (range 24 to 102 months). RESULTS: The mean Constant-Murley score increased from preoperative 41.7 (range 12 to 82) to postoperative 79.7 (range 42 to 100) (p<0.0001). The mean preoperative and postoperative pain scores were 7.5 (range 4 to 10) and 1.25 (range 0 to 5), respectively (p<0.0001). Only eight patients (9%) showed dissatisfaction with the surgical outcome. The remaining 80 patients (91%) were satisfied with the outcome and returned to their previous night comfort and daily activities. None of the patients had postoperative nerve palsy or limitations in shoulder functions. Two patients (2.2%) developed superficial soft tissue infections that disappeared following multiple wound debridements. CONCLUSION: Our long-term results favor open surgical repair and acromioplasty in the treatment of patients with rotator cuff tears. PMID- 15129029 TI - [Late results of arthroscopic and open anterior acromioplasty]. AB - OBJECTIVES: We evaluated the results of arthroscopic or open anterior acromioplasty after a mean follow-up of two years in the treatment of impingement syndrome. METHODS: The study included 145 patients who underwent anterior acromioplasty through open surgery (90 patients; mean age 56 years; range 36 to 70 years) or arthroscopic surgery (55 patients; mean age 49 years; range 32 to 58 years). Comparisons were made between both groups with regard to preoperative and postoperative Constant scores, activity, pain, range of motion, and strength scores, and durations for postoperative pain, hospital stay, and return to work. The mean follow-up was 24 months (range 12 to 60 months) for both groups. RESULTS: The mean Constant scores increased from preoperative 32.6 to postoperative 80.5 and from 25.5 to 83.1 with open and arthroscopic surgery, respectively. No significant differences were found between the two groups with regard to pre- and postoperative Constant scores, activity, pain, range of motion, and strength scores (p>0.05). However, patients who underwent arthroscopic surgery exhibited shorter durations in the following: hospital stay (1.36 vs 1.72 days), delay in return to work (9 vs 16 days), and postoperative pain (6 vs 8 weeks) (p<0.05). According to subjective patient evaluations, arthroscopic results were good or excellent in 47 patients (85.5%), satisfactory in six patients (10.9%), and poor in two patients (4.6%); open surgery results were good or excellent in 76 patients (84.4%), satisfactory in 10 patients (11%), and poor in four patients (4.6%). CONCLUSION: Both treatment options seem to be equally effective in the treatment of impingement syndrome in the absence of rotator cuff tears. PMID- 15129030 TI - [Giant-cell tumors of the tendon sheath involving the hand or the wrist: an analysis of 141 patients]. AB - OBJECTIVES: Giant-cell tumors of the tendon sheath localized in the hand or wrist were retrospectively reviewed with respect to diagnosis, surgical treatment, and postoperative complications. METHODS: During a 21-year period, a total of 141 patients (83 females, 58 males; mean age 37.5 years; range 6 to 77 years) underwent surgery for 146 lesions that developed in the hand (n=134) or the wrist (n=12). Involvement was on the right side in 77 patients, and on the left side in 64 patients. The mean follow-up period was 3.5 years (range 6 months to 11 years). RESULTS: On presentation, the most common symptom was the presence of a painless soft tissue mass. The most frequent localization was the volar part (76%) of the second (27%) and the third (24) fingers, or the proximal phalanx (57%). Forty per cent of tumors were encountered at ages between 30 and 50 years. The duration of symptoms ranged from one month to five years and the highest number of presentations fell within the first six months. In eighteen patients, radiologic studies showed osseous involvement, being cortical sclerosis in 12 patients, and erosion in eight patients. Postoperative complications included digital nerve injuries in four patients, superficial infection in three patients, and joint stiffness in 12 patients. Twenty-three patients (16%) developed recurrences within a mean of 3.7 years (range 2 months to 7 years). CONCLUSION: Taking high rates of recurrences into consideration, surgery for giant-cell tumors of the tendon sheath requires wide surgical exposure, attentive skills, and the use of magnification. PMID- 15129031 TI - [Radiographic and functional results of the Lindgren-Turan operation in the treatment of hallux valgus]. AB - OBJECTIVES: We evaluated the radiographic and functional results of the Lindgren Turan operation in the treatment of hallux valgus. METHODS: Twenty-nine feet of 25 patients (15 women, 7 men; mean age 42 years; range 20 to 68 years) were treated by the Lindgren-Turan method. Of these, 24 feet with appropriate follow up were included. Intermetatarsal and hallux valgus angles were measured on preoperative, postoperative, and follow-up anteroposterior radiographs. Pain was evaluated by a visual analog scale. The range of motion of the first metatarsophalangeal joint was measured at the end of follow-ups. The mean follow up period was 26.5 months (range 6 to 54 months). RESULTS: Postoperatively, the mean corrections were 6.9 degrees and 5.1 degrees in the hallux valgus and intermetatarsal angles, respectively (p<0.05). At the end of the follow-up period, the mean loss of correction was 1.8 degrees for the hallux valgus angle, and 0.5 degrees for the intermetatarsal angle; the mean pain score decreased from preoperative 7.78 to 1.35 (p<0.05) and the mean range of motion of the first metatarsophalangeal joint was 56.8 degrees dorsiflexion and 14.9 degrees plantar flexion. The mean time to return to presurgery activity levels was 44.2 days (range 30 to 70 days). Of the patient group, 83.3% expressed satisfaction with the surgical outcome. Clinical and radiographic evaluations showed excellent results in 15 (62.5%), good in seven (29.2%), and unsuccessful in two patients (8.3%). CONCLUSION: The Lindgren-Turan osteotomy is a preferable method in the surgical treatment of patients with moderate hallux valgus. PMID- 15129032 TI - [The treatment of comminuted midfoot fractures with distraction osteogenesis]. AB - OBJECTIVES: We evaluated the results of distraction osteogenesis with the Ilizarov circular external fixator in the treatment of comminuted fractures of the midfoot. METHODS: Four patients (1 female, 3 males) with comminuted midfoot fractures due to high energy trauma were treated with distraction osteogenesis performed by the Ilizarov circular external fixator. The mean age was 24 years (range 19 to 37 years). Fractures resulted from a traffic accident in one case, and from vehicle blast due to landmine explosion in three cases. The circular external fixator was applied for a mean of three months (range 2.5 to 4 months), during which the mean distraction was 10.5 mm (range 9 to 13 mm) at the fracture zone. The results were assessed according to the presence of pain and limping, radiographic measurements, and to the AOFAS (the American Orthopaedic Foot and Ankle Society) scoring system. The mean follow-up period was 58 months (range 33 to 81 months). RESULTS: All the patients had limping for a month following the removal of the fixator. At the end of six months, limping disappeared in one patient, decreased in two patients, and remained as a sequela in one patient. Pain which occurred during walking after the removal of the fixator decreased and finally disappeared within 3 to 6 months in three patients, but remained in one patient. Radiographic assessments were made using the anteroposterior and lateral X-rays of the foot. The mean AOFAS score was 70.5 (range 50 to 89). CONCLUSION: Distraction osteogenesis performed with the use of the Ilizarov circular external fixator may be an alternative in the treatment of comminuted foot fractures due to high energy traumas, where no other modality is likely to provide an anatomical reduction. PMID- 15129033 TI - [Low back pain among children and adolescents]. AB - OBJECTIVES: We presented our experience with the diagnosis and treatment of low back pain in children. METHODS: We retrospectively reviewed 29 children (15 girls, 14 boys; mean age 12 years; range 9 to 17 years) who were treated for low back pain and had appropriate follow-ups. Etiologic causes were sought by clinical evaluation, radiologic studies, and laboratory tests. Patients with unknown etiology underwent symptomatic treatment. The mean follow-up period was 42 months (range 12 to 96 months). RESULTS: Etiology was determined in 26 patients (89%), which included spondylolysis/spondylolisthesis (n=8), Scheuermann's disease (n=6), neoplasia (n=5), discitis/vertebral osteomyelitis (n=4), and lumbar disc herniation (n=3). Spondylolysis was managed conservatively, except for one patient who had in situ spinal fusion for associated spondylolisthesis. Five patients with Scheuermann's disease were treated conservatively, while one patient required spinal fusion. Surgical treatment with biopsy, curettage, and bone grafting was performed for all neoplasias, but one which was followed-up conservatively. Two patients with discitis were managed with antibiotic treatment and two patients with vertebral osteomyelitis (Pott's abscess) underwent both medical treatment and surgical drainage and stabilization with strut graft. Lumbar disc herniation was treated conservatively in two patients, while one had surgical treatment with excision of disc fragments and limited laminectomy. Finally, all the patients became asymptomatic on final examinations. CONCLUSION: Serious consideration should be given to persistent low back pain in children. Clinical, radiologic, and laboratory findings can be elaborated into etiologic diagnoses and complete relief can be achieved with appropriate treatment. PMID- 15129034 TI - [The assessment of anxiety levels in patients with posttraumatic stress disorder]. AB - OBJECTIVES: The purpose of this study was to measure state and trait anxiety levels of veterans who developed posttraumatic stress disorder following combat or landmine injuries, or vehicle accidents. METHODS: The anxiety levels of 98 veterans (mean age 20 years; range 18 to 25 years) were measured with the use of the State-Trait Anxiety Scale (STAI I-II). Diagnosis of posttraumatic stress disorder was made on the basis of the Structured Clinical Interview for DSM III R. RESULTS: The number of patients with high levels of anxiety was not in significant relationship with education level and trauma type (p>0.05). Regardless of education levels and trauma types, the number of patients with high trait anxiety significantly outweighed the number of patients with high state anxiety (p<0.05). Trait anxiety levels differed significantly between primary school and high school graduates (p=0.03). CONCLUSION: Veterans should be under close supervision to determine and treat increased anxiety which worsens the quality of life. Special attention should be paid to appropriate psychological rehabilitation throughout the treatment plan. PMID- 15129035 TI - [Recurrent traumatic hip dislocation in a child]. AB - Recurrent traumatic hip dislocation is rarely seen in childhood. We presented a case of traumatic hip dislocation which was treated by conservative methods. A two-year-old girl was treated with closed reduction and a hip spica cast for posterior traumatic dislocation in the right hip. Two years later, a recurrent dislocation occurred following a minor trauma on the same side. She was again treated with closed reduction and immobilization with a hip spica cast for three weeks followed by a three-week load bearing restriction. Early follow-up examinations showed an increased internal rotation of the hip compared to the left side, suggesting increased posterior capsular laxity. It then disappeared on her final follow-up at age six and magnetic resonance imaging showed no evidence for avascular necrosis or capsular laxity. She was asymptomatic and her physical examination was within normal limits. Of note, several members in her family had developmental dysplasia of the hip. PMID- 15129036 TI - [Avascular necrosis of the third metacarpal head: a case report]. AB - Avascular necrosis of the third metacarpal head is a rare entity. It may remain asymptomatic, but may also be painful and lead to restricted range of motion. A thirteen-year-old male patient presented with a painful and restricted range of motion in the third metacarpophalangeal (MCP) joint. There was no history of any trauma or a predisposing factor such as systemic lupus erythematosus or steroid use. The range of motion of the involved joint was minimally restricted (flexion range, 10 to 80 degrees). Mild tenderness was found on palpation over the dorsal aspect of the third MCP joint. Radiographs showed flattening and sclerosis of the third metacarpal head together with cystic lesions. The symptoms were controlled with non-steroidal anti-inflammatory drugs and splinting for four weeks. The range of motion of the MCP joint increased, along with regression on radiographs. PMID- 15129037 TI - Sexual health: a useful public health paradigm or a moral imperative? AB - The past decade has shown an increasing use of the concept of sexual health. This upsurge is especially noticeable not only in the field of health education and promotion but also in academic sources. The concept is typically used self evidently and with widely diverse connotations. The definition and understanding of sexual health are still evolving and pose various critical questions. For instance, the term sexual health may imply various risks, including a one-sided health perspective on sexuality and a new excuse to control sexual behavior. The recent discourse on sexual health is paralleled by an upsurge in the debate on sexual rights. Both concepts serve different functions but are intricately interwoven. In this introductory paper, we introduce this Special Section and hope to further the debate and scientific exploration of sexual health. PMID- 15129038 TI - Defining sexual health: a descriptive overview. AB - This paper examines the historical events that have contributed to evolving definitions of sexual health. Through a review of the literature, eight definitions of sexual health were identified from the original 1975 World Health Organization (WHO) definition up until the most recent definition in 2002. Each of these definitions is reviewed for the context in which they were developed and the particular contributions they have played in the development of the understanding of sexual health. The more recent definitions have built upon the essential elements provided in the original WHO definition but have added concepts of mental health, responsibility, and sexual rights. Although similarities exist in the definitions, they are not all alike. The importance of defining sexual health in the discussion of promoting sexual health is illustrated. The concept and definition of sexual health will continue to evolve shaped by historical events. PMID- 15129039 TI - Nonvolitional sex and sexual health. AB - Nonvolitional sex is sexual behavior that violates a person's right to choose when and with whom to have sex and what sexual behaviors to engage in. The more extreme forms of this behavior include rape, forced sex, childhood sexual abuse, sex trafficking, and violence against people with nonconventional sexual identities. More nuanced forms of nonvolitional sex include engaging in sexual behavior that masks one's nonconventional sexual identity, or that protects one's position with peers, or that represents a quid pro quo for the economic support that one obtains within an intimate relationship. The aim of this essay is to highlight the ways in which nonvolitional sex threatens sexual health and to identify strategies for ameliorating this problem. These strategies will have to be as broad in scope as is the problem that they are designed to address. The essay discusses the following strategies to reduce nonvolitional sex: (1) advocacy for sexual rights, gender equality, and equality for individuals with nonconventional sexual identities; (2) primary prevention programs and interventions that offer comprehensive sexuality education that establishes volitional sex and sexual health as basic human rights; (3) health services that routinely ask clients about their experiences with nonvolitional sex in an open and culturally appropriate manner; and (4) secondary prevention programs to meet the needs of victims of nonvolitional sex identified by the "screening" programs. PMID- 15129040 TI - HIV-prevention-related sexual health promotion for heterosexual men in the United States: pitfalls and recommendations. AB - Although the male condom remains the most commonly used method of HIV prevention, sexual health promotion interventions directed toward heterosexuals in the United States have focused primarily on women. In this paper, we discuss limitations of the utility of various HIV-prevention-related sexual health promotion messages as they pertain to the sexual behavior of heterosexual men. We also present several key considerations for the development of sexuality-based HIV health promotion directed toward this population, including the importance of developing HIV risk reduction messages that are responsive to (1) their predominant sexual and safer sex behavioral patterns; (2) the societal gender roles, norms, and scripts that guide heterosexual interactions; and (3) developmental and cultural influences on sexual behavior. We conclude with recommendations for future research. PMID- 15129041 TI - Examining sexual health discourses in a racial/ethnic context. AB - Sexual health is a complex, multidimensional construct. In analyzing race and ethnicity in sexual health, this article examines 2 domains of discourse: (1) preventive sexual health, a public health oriented domain of discourse concerned with the prevention of disease, and (2) eudaemonic sexual health, a domain of discourse concerned with attainment of sexual pleasure within a moral context. Research on the sexual health of ethnic minority populations is typically focused on preventive sexual health. There is an underemphasis on eudaemonic discourses when applied to ethnic minority persons, particularly in the examination of ethnic variations in sexual dysfunction. Medicalization of sexual dysfunction, control of minority sexualities, and ethnically oriented cultural scenarios are cited as reasons for this underemphasis. Exploring racial/ethnic aspects of eudaemonic sexual health can deepen understanding of sexual health in multiple domains and strengthen the ability to promote sexual health for all. PMID- 15129042 TI - A study in sexual health applying the principles of community-based participatory research. AB - The principles of community-based participatory research were applied to an exploratory sexual health study that examined "cruising for sex" among men on a college campus. In the context of a study seeking a broad interpretation of the health implications of cruising, and when faced with methodological challenges, the researchers found these principles to provide invaluable guidance. A review of the research process is offered and the manner in which the principles of community-based participatory research were operationalized for this study is described. PMID- 15129043 TI - Environmental influences on safer sex in young gay men: a situational presentation approach to measuring influences on sexual health. AB - Environmental influences on sexual behavior are difficult to examine given their temporal distance from the sexual act and the cost of long-term longitudinal studies. We examined environmental influences on risky sexual behavior in young gay men using the Situational Presentation (Sitpres) methodology, where situations in which relevant environmental variables are presented as computer vignettes with the variables randomly allocated, and participants rate the likelihood of their engaging in unsafe sexual behavior. A total of 100 gay men aged between 18 and 26 years of age completed 20 situational presentations with the outcome being the likelihood of engaging in unprotected anal intercourse. On regression analysis, 3 environmental variables significantly predicted safer sex: perceived gay/bisexual men's norms toward condom use; availability of HIV prevention messages; and what one's religion says about gay sex. Not significant were family, media, legal, and work/school attitudes to homosexuality. Demographic variables that were predictors included education, age, sexual orientation, and degree of being "out" about sexual orientation. These data suggest that environmental factors can be approximated using the Sitpres methodology, and that more proximal environmental variables have a stronger impact than distal ones. PMID- 15129044 TI - Sex education as health promotion: what does it take? AB - Health promotion should be evidence-based, needs driven, subject to evaluation, and ecological in perspective. How can this be achieved in the context of school based sex education? Adopting new behaviors and giving up old habits involves common decision-making, planning, motivational control, and goal prioritization processes. Consequently, despite the particular nature of sexual behavior, models of cognitive change applied to other health-related behaviors also apply to the promotion of safer sex practice. Young people are less likely to have unprotected sexual intercourse if they have acquired a variety of social skills relevant to dealing with romantic and sexual relationships. Many of these social skills and the methods employed to facilitate their development are also important to the promotion of other health behaviors. However, teaching social skills relevant to sexual behavior in classroom settings requires specialist expertise both in program design and in delivery by teachers or facilitators. Theory- and evidence based programs designed to promote such skills and modified on the basis of effectiveness evaluations are most likely to have an impact on the rate of pregnancies and sexually transmitted infections among young people. Yet, such programs may not be used in schools because of policy and cultural constraints. When this is the case, optimally effective health promotion cannot be delivered. Health promoters should acknowledge these challenges and facilitate the adoption and implementation of effective sexual health promotion programs by targeting communities and legislators. PMID- 15129045 TI - "Let's talk about sex, baby": community-based HIV prevention work and the problem of sex. AB - Talking about sex and sexuality has been integral to HIV/AIDS prevention work in the United States since the beginning of the epidemic. Early prevention workers, who were primarily gay men, developed a "sex-positive" approach to prevention, involving frank discussions of sex and sexuality, with the idea of helping to end the epidemic by protecting oneself and one's sex partners from the disease. This article examines early prevention work at AIDS Action Committee (ACC) in Boston, Massachusetts, and details subsequent challenges to this approach, primarily from Bostonians outside of the gay community, but also from within AAC. The article ends with an examination of the early 2000s at AAC, which saw the more recent manifestation of a "sex-positive" approach to prevention in their work with HIV infected individuals. PMID- 15129046 TI - Sex will never be the same: the contributions of Alfred C. Kinsey. AB - Kinsey built upon what other European and American researchers had done, but in his male volume he was much more critical of his predecessors than he was in the female volume. Although he mentioned many of the European sex researchers, several were conspicuous by their absence and, at times, he seemed very moralistic (e.g., that Hirschfeld was not an objective researcher because of his campaign for gay rights or the failure of H. Ellis to have face-to-face contact with his participants). He had little positive to say about psychiatrists in general, although he imparted a more positive message in the female volume. If bibliographical citations are any example, Kinsey explored much more widely in the social sciences in the female volume than he did in the male volume, indicating that he himself acquired greater expertise over the years. Certainly, the female volume was a more well rounded treatment. Overall, the effect of his books was to change the way people looked at sex; indeed, sex could never be the same again. PMID- 15129047 TI - A phenomenology of problematic sexual behavior occurring in sleep. AB - First-person reports of individuals' experiences of problematic "sleep sex" were collected in an Internet-based study. Qualitative analysis of 121 reports yielded 6 distinct themes: (1) fear and a lack of emotional intimacy; (2) guilt and confusion; (3) a sense of repulsion and feelings of sexual abandonment; (4) shame, disappointment, and frustration; (5) annoyance and suspicion; (6) embarrassment and a sense of "self-incrimination." Results suggest that sleep sex can elicit negative emotions and cognitions that may become a source of personal and relational distress. Clinician familiarity with problematic sleep sex may foster more effective communication with individuals presenting with such complaints. PMID- 15129048 TI - Sexual responsiveness in women with spinal cord injuries: differential effects of anxiety-eliciting stimulation. AB - Sexual dysfunction is a common problem in women after spinal cord injuries (SCIs). Recently, the use of anxiety-provoking stimulation has been explored as a means of improving sexual responses in able-bodied sexually functional and dysfunctional women. In this laboratory-based study, we assessed the sexual and autonomic responses of women with SCIs with varying degrees of preservation of sympathetic innervation to their genitals to respond to anxiety-provoking audiovisual (AV) stimulation. Subjects were 45 women with SCIs and 11 able-bodied women. For purposes of analysis, SCI subjects were grouped on the basis of the degree of preservation of sensation in the T11-L2 dermatomes. Results revealed that women with low sensory scores in these dermatomes achieved higher vaginal pulse amplitude (VPA) responses to audiovisual erotic stimulation after anxiety preexposure than after neutral preexposure; however, women with SCIs and the greatest degree of preservation of sensory function in the T11-L2 dermatomes, as well as able-bodied controls, did not. Moreover, these same 2 groups of subjects had a decrease in VPA responses during baseline periods in which an anxiety provoking video sequence was shown, but not during the neutral sequence. It is concluded that these findings are due to the proximity of sensory and autonomic neurologic elements in the spinal cord. Moreover, they demonstrate the differential effects of sympathetic stimulation on genital sexual arousal. PMID- 15129049 TI - The risk of birth defects in children born after assisted reproductive technologies. AB - PURPOSE OF REVIEW: This review addresses the question of whether there is evidence of an increased risk of birth defects in children born following assisted reproductive technologies compared with spontaneously conceived children. RECENT FINDINGS: Three recent studies added relatively little new information given their modest size (56-472 assisted reproductive technology children). We therefore considered all published papers that compared birth defects in children born following assisted reproductive technologies with those in children born following spontaneous conception. Overall, only six of the 26 relevant papers concluded there was an increased risk of birth defects following assisted reproductive technologies. The interpretation of many studies was based on statistical significance testing alone. When results showed a greater proportion of defects in the assisted reproductive technology group compared with the spontaneous group, but the results were not statistically significant, this was often interpreted as showing no increase in risk, rather than an increase in risk that may have been due to chance. The vast majority of individual studies were too small to have sufficient power to detect, as statistically significant, clinically relevant results. We found that although only eight (30%) of the studies had statistically significant results, 24 (89%) had an odds ratio estimate comparing assisted reproductive technology with spontaneously conceived children of over 1.0; 19 (70%) had an estimate of 1.20 or greater; and 14 (52%) had an estimate of 1.5 or greater. SUMMARY: Current evidence suggests there is an elevated risk of birth defects in children born following assisted reproductive technologies. Whilst others may disagree with our interpretation, one certainly cannot exclude this possibility on the basis of the current evidence. PMID- 15129050 TI - State of the art in in-vitro oocyte maturation. AB - PURPOSE OF REVIEW: The recovery of immature oocytes followed by in-vitro maturation (IVM) and in-vitro fertilization is an attractive alternative to conventional in-vitro fertilization treatment in which controlled ovarian stimulation with gonadotropins is used to increase the number of available oocytes and embryos. Significant progress has been made to improve pregnancy and implantation rates from in-vitro matured oocytes. This review summarizes current knowledge and achievements in human oocyte in-vitro maturation for clinical application, and will highlight recent advances reported in in-vitro maturation treatment. RECENT FINDINGS: It has been demonstrated that priming of ovarian immature oocytes with follicle-stimulating hormone or human chorionic gonadotropin prior to immature oocyte retrieval improves oocyte maturation rates and embryo quality as well as pregnancy rates in infertile women with polycystic ovaries or polycystic ovary syndrome. The size of follicles may be important for the subsequent embryonic development, but the developmental competence of oocytes derived from the small antral follicles is not adversely affected by the presence of a dominant follicle. However oocyte maturation in vitro is profoundly affected by culture conditions. Currently more than 300 healthy infants have been born following immature oocyte retrieval and in-vitro maturation. In general, the clinical pregnancy and implantation rates have reached 30-35% and 10-15% respectively in infertile women with polycystic ovaries or polycystic ovary syndrome. SUMMARY: In-vitro maturation treatment can now be offered as a successful option to infertile women with polycystic ovaries or polycystic ovary syndrome. It is possible to combine natural cycle in-vitro fertilization with immature oocyte retrieval followed by in-vitro maturation, and thus offer women with various causes of infertility reasonable pregnancy and implantation rates without recourse to ovarian stimulation. Further research remains to be done to address the mechanism of oocyte maturation in order to refine culture conditions and improve the implantation rate of oocytes matured in vitro. PMID- 15129051 TI - Evidence-based diagnosis and management of tubal factor infertility. AB - PURPOSE OF REVIEW: The investigation for potential tubal disease is an essential step in the work-up of infertility. This review article provides an evidence based overview of the diagnosis and management of tubal factor infertility. RECENT FINDINGS: While laparoscopic chromopertubation remains the gold standard in the diagnosis of tubal disease and hysterosalpingography is still widely used, newer modalities offer some advantages. Sonohysterography with the use of contrast medium is superior to hysterosalpingography and comparable to laparoscopic chromotubation in diagnosing tubal blockage. Chlamydia serology is the most cost-effective and least invasive diagnostic test for tubal disease, and it is comparable to, if not better than, hysterosalpingography. Depending on the nature and degree of tubal dysfunction as well as the age and ovarian reserve of the patient, various treatments for tubal infertility are available. For proximal tubal obstruction, transcervical tubal cannulation with tubal flushing is a reasonable first approach. Surgical techniques for tubal repair, such as salpingostomy or fimbrioplasty for distal tubal obstruction, can provide good results. Still, tubal factor remains a major indication for in-vitro fertilization and embryo transfer, which bypasses the tubal problem altogether. In certain situations, such as the presence of hydrosalpinx, prophylactic surgery can be used in conjunction with in-vitro fertilization and embryo transfer. SUMMARY: As with infertility in general, the diagnosis and management of tubal infertility should be tailored to the individual patient. Future studies should help to further clarify the role of the various diagnostic tests and therapeutic approaches for tubal infertility. PMID- 15129052 TI - Assessment of ovarian reserve. AB - PURPOSE OF REVIEW: Various methods have been proposed and are currently used in the assessment of ovarian reserve in order to predict the outcome in assisted reproduction. This review focuses on the current information regarding the ovarian reserve markers and tests, an active area of research. RECENT FINDINGS: Female age alone is a rough parameter for assessing ovarian reserve. The basal follicle stimulating hormone level is not adequately sensitive to predict poor outcome and the same is true for other basal parameters, including basal estradiol, the follicle stimulating hormone/luteinizing hormone ratio, and inhibin-B levels. The clomiphene citrate challenge test has a low sensitivity but this sensitivity is greater than that of basal follicle stimulating hormone. The value of other hormonal dynamic tests remains to be determined. Among the ultrasound markers the total antral follicle count seems to be promising. Ovarian biopsy in this assessment is controversial. History of poor response to ovarian stimulation is a strong indicator for declining ovarian function. SUMMARY: There is currently no clinically useful predictive test sufficiently accurate and distinct in time from controlled ovarian stimulation to assess ovarian reserve accurately. Further research is warranted to validate the ovarian reserve tests in the general population and in populations with different causes of infertility. PMID- 15129053 TI - Non-surgical management of leiomyoma: impact on fertility. AB - PURPOSE OF REVIEW: This review is designed to discuss the literature, published from December 2002 to January 2004, on the non-surgical treatment of fibroid. All established and new modalities are reviewed, and all new developments in the field are discussed. The resulting impact on the treatment of infertility will also be evaluated. RECENT FINDINGS: In the past year important advances occurred in the medical treatment of uterine fibroids. It was found, in a large randomized trial, that preoperative treatment with gonadotropin-releasing hormone analogue did not improve surgical results or decrease blood loss. However, important strides were made in understanding the molecular biology of the effect of the hormone analogue on fibroids. Other medical therapies investigated include danazol, raloxifene, mifepristone, aromatase inhibitors, and the levonorgestrel containing intrauterine device. Most promising in terms of long-term usage for reduction of size and symptoms appears to be the combination of gonadotropin releasing hormone analogue and raloxifene, although selective progesterone receptor modulators may also achieve this aim. However, none avoid producing an anovulatory state that inhibits fertility, and none have been shown to enhance fertility following discontinuation. Uterine artery embolization is another non surgical technique under intense investigation. The year's literature suggests that while results are comparable with hysterectomy in terms of complication rate and patient satisfaction, there may be important issues for women who wish to undergo the procedure and retain future fertility. Specifically, there is a significant rate of premature ovarian failure, as well as occasional damage to the endometrial vasculature with resulting atrophy and adhesion formation. Improvements in technique, in particular the use of larger and more spherical microspheres for embolization, may reduce these unwanted effects. However, few data exist regarding the course of pregnancy and outcome following embolization. SUMMARY: A number of non-surgical treatments exist for uterine fibroids, but none has been shown to be of value in the patient desiring future fertility. Myomectomy remains the standard of care for such women, and all other therapies should be designated experimental and limited to appropriate investigational studies. PMID- 15129054 TI - Should we continue or stop insulin sensitizing drugs during pregnancy? AB - PURPOSE OF REVIEW: The use of insulin sensitizing drugs such as metformin in polycystic ovary syndrome has been increasingly popular and validated by systematic reviews. There has also been an interest in the use of metformin for gestational diabetes. However, administration of metformin to prevent miscarriage is controversial and widespread use of this drug in early pregnancy requires investigation. RECENT FINDINGS: There are claims that miscarriage and gestational diabetes are more common in polycystic ovary syndrome and that use of insulin sensitizers improves outcomes dramatically. This review suggests there is no evidence for increased risk of miscarriage solely due to polycystic ovary syndrome and that there are insufficient data for promoting therapy with metformin. There is some reason for use of metformin in mid-pregnancy for gestational diabetes but better evidence from randomized controlled trials is urgently needed. SUMMARY: The use of metformin in early pregnancy for reducing the risk of miscarriage should be avoided outside of the context of properly designed prospective randomized trials. Safety in early pregnancy appears to be reassuring but not completely proven. The use of metformin in mid-pregnancy for gestational diabetes appears more logical but also needs adequate trials before general use is advocated. PMID- 15129055 TI - Single versus double intrauterine insemination: are outcomes affected? AB - PURPOSE OF REVIEW: Intrauterine insemination with or without superovulation is the initial step in assisted reproductive technologies. There have been many attempts to increase the efficiency of this route. One approach may be to increase the frequency of insemination. In the last decade, there has been a continuing debate regarding the increased efficiency of IUI with double IUI. RECENT FINDINGS: Although the initial studies showed that superovulation with double intrauterine insemination had better pregnancy rates than with a single application, recent studies found that ovarian stimulation with double insemination has not increased the pregnancy rates. SUMMARY: Superovulation with intrauterine insemination is a treatment modality used in unexplained infertility and mild male infertility. Increasing the efficiency of the technique has always been an interest of research. Double intrauterine insemination has been suggested to increase efficiency; recent studies, however, have not confirmed this finding. PMID- 15129056 TI - HIV and reproduction. AB - PURPOSE OF REVIEW: Three quarters of individuals infected with HIV are in their reproductive years and can expect an almost normal life expectancy under antiretroviral treatment. Many of them want to have a child and reproductive counselling and care can offer a sharp reduction in both sexual and vertical transmission rates. RECENT FINDINGS: Most couples with HIV are formed by an infected man and an uninfected woman; in this setting, semen washing coupled with reproductive technology can be applied to eliminate the risk of sexual transmission of the virus. Semen washing is a processing method which reduces both HIV RNA and DNA to undetectable amounts. In couples in which only the woman is infected, self-insemination might be indicated. When both partners are carrying HIV, semen washing can be used in couples with different viral strains. HIV can be vertically transmitted and the risk of infection for the infant can be decreased to approximately 1% by reducing maternal viral load, elective caesarean section and avoidance of breastfeeding. In pregnancy the efficacy of antiretroviral treatment should be balanced against the possibility of embryonic or fetal toxicity. Caesarean section, performed electively, has proven its protective efficacy, without significant maternal morbidity. Its role should now be reassessed in mothers with undetectable viral load. Breastfeeding, discouraged to avoid postnatal transmission, might be possible in the future, with antiretroviral therapy capable of suppressing viral excretion in maternal milk. SUMMARY: Semen washing, reproductive technology, antiretroviral therapy and obstetrical care can work in sequence to allow safe reproduction in couples infected with HIV. PMID- 15129057 TI - Bibliography. Current world literature. Fertility. PMID- 15129059 TI - Effects of functional restoration versus 3 hours per week physical therapy: a randomized controlled study. AB - STUDY DESIGN: Randomized parallel-group comparative trial with a 6-month follow up period. OBJECTIVE: To compare, in chronic low back pain patients, the effectiveness of a functional restoration program, including intensive physical training, occupational therapy, and psychological support to an active individual therapy consisting of 3 hours physical therapy per week during 5 weeks. SUMMARY OF BACKGROUND DATA: Controlled studies conducted in the United States showed a benefit of functional restoration in patients with low back pain, especially on return to work. Randomized Canadian and European trials had less favorable results. In France, there has been up to now no randomized study. Controlled studies suggested a positive effect of functional restoration programs. METHODS: Eighty-six patients with low back pain were randomized to either the functional restoration (44 patients) or the active individual therapy (42 patients) program. One person in each group never started the program. Two patients did not complete the functional restoration program, and one was lost to follow-up at 6 months. The mean number of sick-leave days in the 2 previous years was 6 months. RESULTS: After adjustment on the variable "workplace enrolled in an ergonomic program", the mean number of sick-leave days was significantly lower in the functional restoration group. Physical criteria and treatment appreciation were also better. There was no significant difference in the intensity of pain, the quality of life and functional indexes, the psychological characteristics, the number of contacts with the medical system, and the drug intake. CONCLUSIONS: This study demonstrates the effectiveness of a functional restoration program on important outcome measures, such as sick leave, in a country that has a social system that protects people facing difficulties at work. PMID- 15129061 TI - An anatomic and morphometric study of C2 nerve root ganglion and its corresponding foramen. AB - STUDY DESIGN: Exposing and measuring the dorsal root ganglion of the second cervical spinal nerve (C2 ganglion) and the second intervertebral space, which is present between posterior arch of atlas (APA) and lamina of axis (LA). OBJECTIVES: This study aims to investigate the shape, size, and relation of the C2 ganglion with the adjacent structures that limits the corresponding intervertebral space and the alterations of relation between C2 ganglion and APA and between C2 ganglion and LA with the movements of the head bilaterally. SUMMARY OF BACKGROUND DATA: In previous studies, the position and the heights of the C2 ganglion have been described. But the shape of the C2 ganglion and its relation to APA and LA by the movement of the head had not been considered previously. METHODS: Upper cervical spines of 20 cadavers were dissected posteriorly. The muscles attaching to the atlas and axis were resected to ease the head movements. The heights of the C2 ganglion and space were measured in anatomic position and in hyperextension with opposite rotation position of the head. Originally in this study, plastic dough casts were used to obtain reliable outcomes. RESULTS: The shape of the ganglions was defined in three types: 70% were oval, 20% were spindle-like, and 10% were spherical. The height of the C2 ganglion was 4.97 +/- 0.92 mm on the right side and 4.6 +/- 0.84 mm on the left side. The height of the intervertebral space in anatomic position and in hyperextension with rotation to the opposite position of the head were, respectively, 9.74 +/- 1.77 mm and 7.48 +/- 1.44 mm on the right side and 9.64 +/ 1.47 mm and 7.12 +/- 0.96 mm on the left side. There was no bone contact or impact to the ganglion in each position of the head. CONCLUSION: The C2 ganglions are confident in their place between APA and LA. No bone contact to the C2 ganglion was detected in either normal limited or in forced head motions. PMID- 15129062 TI - Posterior screw placement on the lateral mass of atlas: an anatomic study. AB - STUDY DESIGN: An anatomic study evaluating the feasibility of posterior screw fixation on the lateral mass of the atlas and comparing the pullout force of C1 posterior lateral mass screws with that of occipital and C2 pedicle screws. OBJECTIVES: To study the feasibility, anatomy, and biomechanics of posterior screw fixation within the lateral mass of the atlas. SUMMARY OF BACKGROUND DATA: Occipitocervical fusion is a common choice for atlantoaxial dislocations. After that the movement of occipitocervical region will be restricted. If screws could be placed in the lateral mass of the atlas, 1) the movement of C0-C1 junction will be preserved; 2) through the screw the dislocated atlas could be pulled backward; and 3) the fractured lateral mass can be fixed using the posterior lateral mass screw directly. METHODS: Thirty atlas specimens of native adults were used to measure pertinent clinical data. Six fresh upper cervical specimens were used to observe the association of the C2 nerve and the lateral mass of the atlas. The pullout force of the C1 lateral mass was measured and compared with that of screws placed into the occiput and C2 pedicle, respectively. RESULTS: The distances from the midline to the midpoint of the C1 lateral mass, to the inner wall of the transverse foramen, and to the inner edge of the pedicle were 17.6 +/ 1.2 mm, 23.0 +/- 1.7 mm, and 14.2 +/-1.1 mm, respectively. The width and height of the C1 lateral mass were 11.6 +/- 1.4 mm and 12.7 +/- 1.0 mm, respectively. The distance between the inferior midpoint of the C1 lateral mass in the transverse plane of the midposterior arch of the atlas to the edge of the inferior atlas joint line was 4.1 +/- 0.7 mm. The width of the posterior arch at the point of vertebral artery overpass was 4.7 +/- 1.0 mm. The observation on the fresh specimens showed that the C2 nerve passes inferolateral to the C1-C2 joint and could be pulled sideways. The screw pullout force within the lateral mass of the atlas was 1818.16 +/- 422.67 N. This is significantly less than that of screws within the occiput, and there was no difference with that of a C2 pedicle screw. CONCLUSION: The width and height of the atlas lateral mass were larger than that of the C2 pedicle, and there was enough space to insert a 3.5-mm diameter screw in the atlas lateral mass over the C2 nerve. The pullout force of the screw on the lateral mass of the atlas was the same as that of the C2 pedicle screw. It is possible toinsert a 3.5-mm screw in the lateral mass of the atlas. The direction of the screw should be about 20 degrees anterosuperior in the vertical plane and 15 degrees inward in the horizontal plane. The suitable length of the screw should be approximately 22 mm inside the lateral mass. PMID- 15129063 TI - Ne-Osteo bone growth factor for posterolateral lumbar spine fusion: results from a nonhuman primate study and a prospective human clinical pilot study. AB - STUDY DESIGN: Prospective animal and human clinical pilot trial. OBJECTIVES: The purpose of this study was to determine and test the dose of Ne-Osteo growth factor extract and carrier required for consistent radiographic bone induction in humans. SUMMARY OF BACKGROUND DATA: Preclinical studies have demonstrated that Ne Osteo, an extract-containing bone morphogenetic proteins, was successful at generating spine fusion in rabbits and rhesus monkeys. Consistent fusions have yet to be achieved in nonhuman primates and humans. METHODS: Adult rhesus monkeys underwent single-level posterolateral intertransverse lumbar arthrodesis with either 3.0 mg (N = 4), 5.0 mg (N = 4), 12.5 mg (N = 4), or 25 mg (N = 4) of Ne Osteo per side. Animals were killed after 24 weeks. In the human clinical trial, 22 patients (18 females, 4 males) had lumbar spinal stenosis and/or spondylolisthesis requiring spine arthrodesis. To minimize patient risk of nonunion, patients received autogenous bone graft from the posterior iliac crest on one side and Ne-Osteo growth factor on the other. The dose was 12.5 mg, 25 or 50 mg, or 25 mg Ne-Osteo per side performed in the three phases, respectively. RESULTS: Three of four monkeys that received 12.5 mg Ne-Osteo per side and four of four that received 25 mg per side achieved solid fusions. In phase I of the human clinical trial, two of six patients showed radiographic bone induction (plain radiograph, CT scans-blindly evaluated) on the Ne-Osteo side (12.5-mg dose). In phase II, both sides were graded as fused in five of six patients. Although graded as fused, the 6-month scans demonstrated a ring of new bone with the center filling in slower (12-24 mo) than was predicted by nonhuman primate studies. As a result, phase III carrier was designed to have a more porous/open early fusion mass than with the dense DBM paste (used in phase I and II) by mixing in local bone or cancellous allograft chips. Results using the 25- and 50 mg doses were the same, so 25 mg was used in phase III. In phase III, 9 of 10 autograft were fused by 12 months. Five of five patients with Ne-Osteo plus local bone and four of five with allograft chips were fused by 6 months. The one patient in this group that did not heal on either the autograft or the Ne-Osteo side was a smoker. CONCLUSIONS: A graft composite of Ne-Osteo bone growth factor with human DBM with or without cancellous allograft or local bone autograft was capable of achieving a contiguous spine fusion mass in 15 of 16 patients at a dose of at least 25 mg per side. This result was comparable with the results using iliac crest autograft (94%) in this side-by-side model. These results warrant confirmation in a definitive trial using Ne-Osteo on both sides of the spine and thus avoiding the need for iliac crest bone graft harvest. PMID- 15129064 TI - Preclinical evaluation of a poly (vinyl alcohol) hydrogel implant as a replacement for the nucleus pulposus. AB - STUDY DESIGN: An in vivo investigation into the safety of a novel hydrogel implant designed to replace the diseased nucleus pulposus. OBJECTIVES: To determine the local and systemic safety of this new implant in a nonhuman primate model. SUMMARY OF BACKGROUND DATA: A poly (vinyl alcohol) (PVA) hydrogel has been developed as a prosthetic replacement for the diseased nucleus pulposus. METHODS: PVA implants were inserted into discectomy defects created in the L3-L4 or L4-L5 intervertebral disc in 20 male baboons. Empty discectomy defects served as a surgical control in 8 additional animals. Routine follow-up evaluations included radiography, magnetic resonance imaging, gross pathology, and histopathology of both local and remote tissues. RESULTS: Insertion of the PVA hydrogel from an anterior direction produced extrusions in 5 animals from the first series of 15 surgeries (33%). A modified surgical technique, involving an anterolateral rather than anterior approach, was used in 5 animals, but the extrusion rate remained high (20%). Despite these surgical complications, the PVA implants were well tolerated over 24 months in vivo, with no evidence of device-related pathology in the adjacent disc tissue, spinal cord, or remote tissues. CONCLUSION: Implantation of the PVA implant for periods of up to 24 months produced no evidence of local or systemic toxicity. Additional studies are now needed to determine the efficacy of the device in its intended application. PMID- 15129065 TI - Vertebral forward slippage in immature lumbar spine occurs following epiphyseal separation and its occurrence is unrelated to disc degeneration: is the pediatric spondylolisthesis a physis stress fracture of vertebral body? AB - STUDY DESIGN: Radiographic and histologic evaluation of a rat model of lumbar spine slippage. OBJECTIVES: To clarify the pathomechanism of slippage in the immature spine. SUMMARY OF BACKGROUND DATA: There are controversial hypotheses regarding the pathogenesis of slippage of the pediatric spine with pars defects. Some studies supported that disc degeneration was its cause, while others indicated the growth plate injury was the cause. METHODS: An immature lumbar spine slippage model in 4-week-old rats was used. Following posterior destabilizing surgery, the lumbar spine was radiographically and histologically examined at 1, 3, 5, and 7 days after surgery. RESULTS: Radiographically, slippage occurred about 7% in the % slip on day 7, and no slippage was observed before day 5. Histologically, epiphyseal separation also appeared on day 7; before day 5, the growth plate showed no abnormalities. Within 7 days after the operation, the anulus fibrosus did not show any sign indicating degeneration. The nucleus pulposus was also normal up to day 7. CONCLUSION: The findings of this study support the hypothesis that vertebral forward slippage of the immature spine occurs following epiphyseal separation and its occurrence is unrelated to disc degeneration. PMID- 15129066 TI - Outcome of pediatric patients with severe restrictive lung disease following reconstructive spine surgery. AB - STUDY DESIGN: Retrospective chart review. OBJECTIVES: To assess outcome and complications in pediatric patients with a vital capacity < or =45% of predicted who underwent reconstructive spine surgery for severe thoracic scoliosis. SUMMARY OF BACKGROUND DATA: Despite the known sequelae of untreated spine deformity, pediatric patients with severe restrictive lung disease are frequently not considered good operative candidates for surgical treatment on the basis of their underlying lung disease. Only a few reports have examined surgical treatment in this population. METHODS: A retrospective chart review was conducted on 21 pediatric patients with restrictive lung disease (average age 11.5 years, range 2 21 years). Eighteen patients completed pulmonary function testing, all with vital capacity < or =45%. The remaining three patients had clinical features of restrictive lung disease but could not perform pulmonary function testing. Eighteen patients underwent combined anterior and posterior spinal fusions. Before surgery, all patients were seen by the pulmonary, cardiology, pediatric, and anesthesia services, as well as neurology and gastroenterology services if indicated. Outcome variables examined were as follows: 1) days on oxygen, 2) need for tracheostomy, 3) days on ventilatory support, 4) mortality rate, 5) pulmonary complications, 6) blood loss, 7) other complications relative to the surgical procedure, and 8) length of stay. RESULTS: For the 21 patients, the preoperative average vital capacity was 32% predicted (range 18-43%). After surgery, the median time on supplemental oxygen was 3 days (range 0-90 days). Only one patient with a vital capacity of 29%, who was successfully extubated on postoperative day 4, developed a deep wound infection, which required incision and drainage, as well as recurrent right lung collapse, and subsequently required a tracheostomy to facilitate ventilator weaning. Four patients were on assisted ventilation before the surgery and continued to require it after surgery. Two additional patients required noninvasive positive pressure ventilation for a short period of time following the surgery. Pneumonia occurred in two patients, and pleural effusion occurred in two patients. Four patients developed postoperative atelectasis. There was one reoperation for implant failure. There were no mortalities or adverse neurologic outcomes. Major thoracic scoliosis measured an average of 82 degrees (range 40-140 degrees) before surgery, with correction to an average of 35 degrees (range 6-75 degrees). CONCLUSIONS: Reconstructive spine surgery in pediatric patients with severe restrictive lung disease and significant spinal deformity is well tolerated. Familiarity with different surgical techniques of salvage reconstruction and perioperative multidisciplinary management should be emphasized. Routine preoperative tracheostomy is not indicated. PMID- 15129068 TI - Selective thoracic fusion for adolescent idiopathic scoliosis with C modifier lumbar curves: 2- to 16-year radiographic and clinical results. AB - STUDY DESIGN: Retrospective clinical and radiographic review with functional outcome assessment. OBJECTIVES: To evaluate outcome of selective thoracic fusion for adolescent idiopathic scoliosis in the presence of widely deviated compensatory lumbar curves. SUMMARY OF BACKGROUND DATA: Previous reports on the results of selective thoracic scoliosis fusion have not specifically focused on deformities with widely deviated lumbar curves. Whether these challenging deformities are best treated with selective thoracic fusion or fusion of both curves remains unclear. METHODS: Forty-four consecutive patients with adolescent idiopathic scoliosis with main thoracic, compensatory minor lumbar "C" modifier curves underwent selective thoracic fusion at a single institution (1987-2000). Radiographs were analyzed before surgery, at 1 week, 2 years, and latest follow up (2-16 years; mean 5.0 years). RESULTS: A mean 36% thoracic correction was closely matched by a 34% lumbar correction at latest follow-up. A majority of spontaneous lumbar correction occurred at its cephalad segments (P = 0.001). Spontaneous correction of lumbar apical translation occurred in a majority of patients (prognostic factors identified). Global coronal imbalance (2-5 cm) was common before surgery and was a significant risk factor (P = 0.04) for global imbalance at latest follow-up. Postoperative bracing was not utilized, and there were no reoperations. Patients with coronal imbalance (2-5 cm) at latest follow up had slightly inferior SRS-24 results. CONCLUSIONS: Satisfactory results are achieved with selective thoracic fusion of properly selected C modifier lumbar curves. Correction of the lumbar curve results principally from a decrease in the tilt of its upper vertebrae, but not necessarily improved apical translation. Mild coronal imbalance was well tolerated and has not necessitated distal extension of the fusion. PMID- 15129069 TI - The course of sagittal plane abnormality in the patients with congenital scoliosis managed with convex growth arrest. AB - SUMMARY OF BACKGROUND DATA: Patient age; localization, length, and magnitude of the curve; and sagittal plane alignment are reported to be the major determinants in the selection of patients for convex growth arrest. Although the existence of sagittal plane abnormality (kyphosis or lordosis) is accepted as a contraindication for convex growth arrest, this issue has not been discussed in detail. OBJECTIVES: The purposes of this study are to investigate the effect of sagittal plane abnormality on the control of coronal plane deformity and to evaluate the course of sagittal plane abnormality of the patients with congenital scoliosis who were satisfactorily managed with convex growth arrest. STUDY DESIGN: Retrospective analysis. METHODS: Inclusion criteria are: 1) a diagnosis of congenital scoliosis in a patient younger than 6 years of age, 2) treatment with convex growth arrest, 3) follow up for more than 2 years, 4) stabilized or improved coronal plane deformity, and 5) abnormal sagittal plane alignment within the scoliotic segment before surgery. The patients were evaluated with anteroposterior and lateral radiographs, and segmental measurements were compared according to the normal of their corresponding age. RESULTS: A total of 38 patients with congenital scoliosis treated with convex growth arrest were reviewed. Among 13 patients with segmental sagittal plane deformity, 2 were excluded because of insufficient control of the scoliosis. Eleven patients (8 girls, 3 boys) with a mean age of 35 months (range 6-72 months) and mean follow up of 40 months (range 24-76 months) fulfilled these criteria. The coronal plane deformities were 58 degrees (range 36 degrees-105 degrees) before surgery and 52 degrees (13 degrees-107 degrees) at the final follow-up. While six of the curves improved, the remaining ones stabilized. Sagittal segmental alignments within the scoliotic segments were hyperkyphotic in 9 patients and hypokyphotic in 1 and lordotic in 1. At the end of the follow-up, sagittal Cobb angle of the abnormal segments remained stable in 7 patients and deteriorated in 4. None of the 4 patients required any reconstructive spine procedure for kyphosis during follow up. CONCLUSION: Sagittal segmental abnormality does not have a negative effect on the control of scoliosis in the majority of the patients (11 of 13). If the coronal curve stabilizes or improves, then sagittal segmental abnormality could also be stabilized (in 7 of 11 patients). PMID- 15129071 TI - Analysis of screw placement relative to the aorta and spinal canal following anterior instrumentation for thoracic idiopathic scoliosis. AB - STUDY DESIGN: Axial computed tomographic (CT) evaluation of the position of anterior vertebral body screws placed thoracoscopically in patients with adolescent idiopathic scoliosis (AIS). OBJECTIVE: To evaluate the position of the anterior vertebral body screws relative to the spinal canal and the thoracic aorta following anterior spinal fusion and instrumentation for AIS. SUMMARY OF BACKGROUND DATA: Thoracoscopic anterior instrumentation and fusion is gaining more widespread use in the treatment of idiopathic scoliosis. However, the accuracy in the positioning of instrumentation has not been previously studied for this technically difficult surgery. METHODS: CT examinations were performed following thoracoscopic anterior spinal fusion and instrumentation in 14 patients with right thoracic AIS. The vertebral body width (transverse dimension) and depth (anterior-posterior dimension) was measured for each thoracic vertebra. At each instrumented level, the position of the screw was analyzed to determine its proximity to the spinal canal and the aorta. The distance from the anterior cortex of the spinal canal to the posterior edge of the screw was measured. The position of each screw relative to the aorta was determined: D, the screw tip was distant to the aorta; A, the screw tip was adjacent to the aorta; C, the screw tip was felt to be against the aorta and creating some contour deformity on the outer wall of the aorta. RESULTS: All 14 patients were female and had a single right thoracic curve. The average age of the patients was 13.3 years (range 12.4 15.1 years). The average preoperative coronal Cobb measurement was 55.9 degrees (bending 26.4 degrees) with correction to 8.9 degrees at 2 years after surgery. The average number of levels fused was 6.6 (range 5-8) and a total of 106 screws were used (average 7.6/patient). The width of the vertebral bodies increased from T4 (24.0 mm) to T12 (33.6 mm), increasing an average of 1.2 mm per level, while the depth increased from 17.7 mm at T4 to 25.5 mm at T12. The average distance from the posterior aspect of the screw to the spinal canal was 5.3 mm (range-1.2 to 11.4 mm). There were no neurologic deficits in any patient. When analyzing the position of the screw tip relative to the aorta, 78 (73.6%) screws were distant from the aorta, 15 (14.2%) were adjacent to the aorta, and there were 13 (12.3%) screws that were thought to create a contour deformity of the aorta. There were no vascular complications at 2 years after surgery. CONCLUSIONS: Thoracoscopic instrumentation and fusion is technically demanding and relies on adequate visualization for accurate screw placement. The vertebral body width and depth are consistent between patients, with the vertebral body width increasing approximately 1.2 mm when progressing down the thoracic spine. Safe screw placement was achieved with adequate distance from the spinal canal; however, close screw proximity to the aorta was seen. The aorta was positioned on the left lateral aspect of the vertebral body in these patients, making anterior screw placement challenging in right thoracic AIS. PMID- 15129073 TI - A prospective comparison of the coronal deformity correction in thoracic scoliosis using four different instrumentations and the fulcrum-bending radiograph. AB - STUDY DESIGN: A prospective study on comparing coronal deformity correction in thoracic scoliosis using four different instrumentations. OBJECTIVES: To compare the ability of four different instrumentation systems in correcting thoracic scoliosis based on the curve flexibility as reviewed by the fulcrum-bending radiograph. SUMMARY OF BACKGROUND DATA: The fulcrum-bending radiograph has been shown to be able to accurately reflect the flexibility of thoracic curves, and the fulcrum bending correction index (FBCI) predicts the amount of correction achieved by current surgical techniques. By recruiting curves of known flexibility, the efficacy of the different instrumentations in correcting coronal deformity can be truly compared. METHODS: A consecutive series of 127 patients with idiopathic scoliosis were treated by one of four implants: CD-Horizon (CD H), Moss Miami (MM), TSRH, and ISOLA. All surgeries were performed by the same group of surgeons using the respective recommended techniques. FBCI was used to compare the correction achieved by these implants. RESULTS: The mean FBCI/correction rate was 101.0%/57.9% in the TSRH group, 103.5%/58.5% in ISOLA, 109.1%/67.6% in CD-H, and 100.2%/62.7% in the MM group. The correction rate was significantly (P < 0.05) higher in the CD-H group than those in the TSRH and ISOLA groups, while the differences in the FBCI between the four implants were not statistically significant. CONCLUSIONS: When curve flexibility is taken into account, despite differences in material and design of four commonly used instrumentations, their ability to correct thoracic scoliosis is the same. Future studies describing surgical correction results should be based on the FBCI. PMID- 15129074 TI - Weight-bearing radiographs in thoracolumbar fractures: do they influence management? AB - STUDY DESIGN: Prospective observational study. OBJECTIVE: Our objective was to compare supine and erect (weight-bearing) radiographs in patients with thoracolumbar fractures without a neurologic deficit and to determine whether the erect radiographs alter the deformity and the management plan. SUMMARY OF BACKGROUND DATA: Nonoperative treatment for thoracolumbar fracture without a neurologic deficit is safe and effective. There are some guidelines in the literature that provide objective standards to identify the patients that are suitable for nonoperative treatment. These guidelines are based on measurements on supine radiographs. The role of weight-bearing radiographs in influencing the management plan of these injuries has not been explored. METHODS: Fractures between T11 and L2 in 28 patients were considered suitable for nonoperative treatment initially. Radiographic measurements included anterior and posterior vertebral body heights, interpedicular distance, and the Cobb angle on the supine and erect radiographs. A change in the treatment from the initial nonoperative management plan, based on the radiographic findings, was recorded. RESULTS: Mean supine Cobb angle of 11 degrees increased to 18 degrees on weight-bearing films. The mean anterior vertebral compression increased from 34% to 46%. No change was noted between the posterior vertebral heights and the interpedicular distance. Seven of the 28 patients were subjected to surgical stabilization based on these findings. CONCLUSION: Performing erect radiographs in patients with thoracolumbar fractures without a neurologic deficit provides additional information and did alter the management plan in a significant proportion (25%) of our patients. PMID- 15129075 TI - Effect of nicotine on spinal disc cells: a cellular mechanism for disc degeneration. AB - STUDY DESIGN: Experimental investigation to determine the effect of nicotine on intervertebral spinal disc nucleus pulposus (NP) cells cultured in vitro. OBJECTIVES.: To evaluate the effects of nicotine on cell proliferation, extracellular matrix production, and viability of NP cells in three-dimensional alginate constructs cultured in vitro. SUMMARY OF BACKGROUND DATA: Numerous studies confirm that smoking is a strong risk factor for back pain. The most widely accepted explanations for the association between smoking and disc degeneration is malnutrition of spinal disc cells by carboxy-hemoglobin-induced anoxia or vascular disease. Nicotine, a constituent of tobacco smoke, present in most body fluids of smokers is known to have detrimental effects on a variety of tissues. It may also be directly responsible for intervertebral disc (IVD) degeneration by causing cell damage in both the nucleus pulposus and anulus fibrosus. The effect of nicotine on IVD cells has not previously been investigated. METHODS: Bovine chondrocytic intervertebral disc cells were isolated by sequential digestion of nucleus pulposus and seeded in 2% alginate. The constructs were cultured for 21 days either in growth medium containing freebase nicotine (Sigma) at concentrations found in the serum of smokers (25 nmol/L-300 nmol/L) or in standard nicotine free-medium as controls. Samples were collected at time points 3, 7, 14, and 21 days and a quantitative assay was performed for DNA, glycosaminoglycans (GAG), and hydroxyproline. Samples were also processed for qualitative histologic analysis including immunolocalization of collagen types I and II. RESULTS: There was both a dose- and time-dependent response to nicotine, with constructs cultured in low-nicotine concentration media demonstrating an early increase in DNA, GAG, and collagen content, while constructs cultured in high nicotine concentration media demonstrated a late decrease in these parameters. At 25 nmol/L dose of nicotine, there was a significant increase (P < 0.05) in the above parameters at day 7 compared with the controls. At higher doses, there was a significant dose-dependent decrease (P < 0.05) in these parameters compared to controls; however, this was only significant at day 14 for the 300 nmol/L group and at day 21 for the 100 nmol/L, 200 nmol/L, and 300 nmol/L groups. Adverse morphologic changes were observed on histology, which included reduced cell proliferation, disrupted cell architecture, disintegration of cells, and extracellular matrix. Immunohistochemistry revealed the presence of type I collagen in the extracellular matrix rather than the normal type II collagen seen in the controls. CONCLUSIONS: Nicotine has an overall detrimental effect on NP disc cells cultured in vitro. There was significant inhibition of cell proliferation and extracellular matrix synthesis. Nicotine in tobacco smoke may have a role in pathogenesis of disc degeneration. PMID- 15129076 TI - Back and neck pain exhibit many common features in old age: a population-based study of 4,486 Danish twins 70-102 years of age. AB - STUDY DESIGN: Cross-sectional and longitudinal analysis of data comprising 4486 Danish twins 70-102 years of age. OBJECTIVES: To describe the 1-month prevalence of back pain, neck pain, and concurrent back and neck pain and the development of these over time, associations with other health problems, education, smoking, and physical, and mental functioning. SUMMARY OF BACKGROUND DATA: Back pain and neck pain are prevalent symptoms in the population; however, there is little research addressing these conditions in older age groups. METHODS: Extensive interview data on health, lifestyle, social, and educational factors were collected in a nationwide cohort-sequential study of 70+-year-old Danish twins. Data for back pain, neck pain, lifetime prevalence of a comprehensive list of diseases, education, and self-rated health were based on self-report. Physical and mental functioning were measured using validated performance tests. Data including associated factors were analyzed in a cross-sectional analysis for answers given at entry into the study, and longitudinal analysis was performed for participants in all four surveys. RESULTS: The overall 1-month prevalence for back pain only was 15%, for neck pain only 11%, and for concurrent back and neck pain 11%. The prevalence varied negligibly over time and between the age groups, and 63% of participants in all surveys had no episodes or only one episode of back or neck pain. Back pain and neck pain were associated with a number of other diseases and with poorer self-rated health. Back and neck pain sufferers had significantly lower scores on physical but not cognitive functioning. CONCLUSIONS: Back pain and neck pain are common, intermittent symptoms in old age. Back pain and neck pain are associated with general poor physical health in old age. PMID- 15129077 TI - Validation of the Turkish version of the Oswestry Disability Index for patients with low back pain. AB - STUDY DESIGN: Validation of a translated, culturally adapted questionnaire. OBJECTIVES: To translate and culturally adapt the Turkish version of the Oswestry Disability Index (ODI) (2.0), and to validate its use for assessing disability in Turkish patients with low back pain. SUMMARY OF BACKGROUND DATA: The ODI is a reliable evaluation instrument for disability, but no validated Turkish version is available. METHODS: A total of 95 outpatients with low back pain were assessed by the ODI. Sixty-five of these patients were observed on a second occasion. Translation/retranslation of the ENG version of the ODI was done blindly and independently by four different individuals, and adapted by a team. Individuals were given the ODI and other scales (Visual Analog Scale, Schober Test, and the Roland-Morris Disability Questionnaire) on their first visit and a week later. RESULTS: Scores of the two ODIs were 27.10 (SD 16.22) on day 1 and 22.88 (SD 13.94) on day 7, with an intraclass correlation coefficient of r = 0.938 (P < 0.001). Cronbach's alpha was 0.918 (day 1) and 0.895 (day 7) in the validation. Concurrent validity, measured by comparing ODI responses with the results of Visual Analog Scale and Schober test, was r = 0.367 (P < 0.01), r = -0.068 (P = 0.591) for day 1, and r = 0.392 (P < 0.01), r = -0.041 (P = 0.745) for day 7, respectively. Construct validity, tested by determining the correlation between the Turkish ODI and the Turkish adaptation of the Roland-Morris Disability Questionnaire, yielded r = 0.815 (P < 0.001) on day 1 and r = 0.708 (P < 0.001) on day 7. CONCLUSION: The Turkish version of ODI has good comprehensibility, internal consistency, and validity and is an adequate and useful instrument for the assessment of disability in patients with low back pain. PMID- 15129079 TI - A low back-specific version of the SF-36 Physical Functioning scale. AB - STUDY DESIGN: A prospective repeated measures design was used to produce a back specific version of the Short Form-36 Physical Functioning scale (SF-36 PF) by Rasch analysis of a pool of items from the SF-36 PF, Oswestry Disability Questionnaire, and the Quebec Back Pain Disability Scale. OBJECTIVES: To identify items for a back-specific version of the SF-36 PF scale and to compare the psychometric properties of the new version with the original 10-item scale. SUMMARY OF BACKGROUND DATA: Adequate assessment of patient function requires the administration of a generic and a condition-specific questionnaire. A back specific version of the SF-36 PF would facilitate comprehensive patient assessment in the clinical setting. METHODS: Consecutive patients with low back pain presenting for physiotherapy treatment were recruited at three public hospitals, three community health services, and four private practices. Patients completed questionnaires on two occasions 6 weeks apart. RESULTS: A scale of 18 items showed a better fit to the Rasch model than the original SF-36 PF scale. Items in the original scale that had a poor fit (INFIT/OUTFIT statistics outside the range 0.7-1.3) showed an acceptable fit in the new scale. The augmented scale had comparable reliability and improved responsiveness to the original 10-item SF 36 PF scale. The minimum detectable change (90% confidence) and the minimum clinically important difference were 12 points. Floor and ceiling effects were practically eliminated. The psychometric properties of the new scale were comparable to those of the Oswestry Disability Questionnaire. CONCLUSIONS: The Low-Back SF-36 PF18 comprises the 10-item SF-36 PF scale and four items each from the Oswestry and Quebec back pain questionnaires. The possible total score ranges from 0 to 100, with a higher score indicating better function. The new scale appears to offer advantages over the use of the original scale for the assessment of functioning in patients with low back pain. PMID- 15129080 TI - Re: Bouter L, Pennick V, Bombardier C. Cochrane Back Review Group. Spine. 2003;28:1215-1218. PMID- 15129082 TI - Re. Kleinstueck FS, Diederich CJ, Nau WH et al. Temperature and thermal dose distributions during intradiscal electrothermal therapy in the cadaveric lumbar spine. Spine 2003;28:1700-8. PMID- 15129084 TI - Re. Kleinstueck FS, Diederich CJ, Nau WH et al. Temperature and thermal dose distributions during intradiscal electrothermal therapy in the cadaveric lumbar spine. Spine 2003;28:1700-8. PMID- 15129085 TI - Re: Foley KT, Holly LT, Schwender JD. Minimally invasive lumbar fusion. Spine 2003;28:S26-35. PMID- 15129086 TI - Diagnostic value of 99mTc-HMDP bone scan in atypical osseous tuberculosis mimicking multiple secondary metastases. AB - STUDY DESIGN: A case of atypical osseous tuberculosis (TB) mimicking multiple secondary metastases on radiologic and nuclear imaging is presented. OBJECTIVES: To emphasize the contribution of nuclear bone scanning for the assessment of osseous tuberculosis in typical and atypical presentations. SUMMARY AND BACKGROUND DATA: Skeletal locations of TB mostly involve the dorsolumbar spine and diagnosis is often delayed. The presence of multiple TB sites can mimic secondary metastases and biopsy remains the mainstay for final diagnosis. METHODS: Clinical symptoms, lab tests, and imaging data are presented. Possible diagnoses are discussed. A review of imaging characteristics in cases of typical and atypical presentations of osseous TB is proposed. RESULTS: A dorsal spine spondylitis was first diagnosed on a 56-year-old patient presenting neurologic deficit of the left arm. Fine needle aspiration identified bacterial infection but was negative for Mycobacterium tuberculosis. Whole-body bone scan allowed the identification of an asymptomatic sacroiliac lesion, which was accessible to biopsy and gave a final diagnosis. CONCLUSION: Nuclear bone scanning should be kept in mind when assessing spinal pain in patients at high risk of TB infection or reactivation. PMID- 15129087 TI - Primary hydatid disease of sacrum affecting the sacroiliac joint: a case report. AB - STUDY DESIGN: A case report of hydatid disease of the spine. OBJECTIVE: To describe an unusual case of hydatid disease of the sacrum affecting the sacroiliac joint and to discuss imaging, differential diagnosis, and treatment. SUMMARY OF BACKGROUND DATA: Hydatidosis or echinococcosis affecting the spine is rare and has a characteristic geographic distribution. Signs of sacroiliac joint involvement and accompanying neurologic deficits cause difficulties in differential diagnosis of this rare condition. METHODS: A case of 38-year-old female patient with low back pain and sciatica was presented. RESULTS: Plain radiographs, computed tomography, and magnetic resonance imaging scans revealed destructive expansive lesion located on the right sacrum and extended through the right sacroiliac joint. Surgical enucleation of the cysts was performed together with mebendazole treatment and histopathologic examination confirmed hydatidosis. CONCLUSION: This unusual disease should be kept in mind in the differential diagnosis of sacroiliac pain and sciatica, especially in endemic areas. PMID- 15129089 TI - Craniovertebral instability with spinal cord compression in a 17-month-old boy with Sly syndrome (mucopolysaccharidosis type VII): a surgical dilemma. AB - STUDY DESIGN: A case study with review of surgical technique in craniovertebral stabilization for young children with mucopolysaccharidosis. OBJECTIVES: To describe an interesting patient with a rare metabolic disorder and review surgical technique for craniovertebral instability in this rare patient population. SUMMARY OF BACKGROUND DATA: Craniovertebral instability has been reported in patients with mucopolysaccharidosis and poses a problem for spinal surgery because of the inherent metabolic disorder and age at presentation. We present the first case of craniovertebral instability and spinal cord compression occurring in Sly syndrome (mucopolysaccharidosis type VII) who is the youngest patient afflicted with this metabolic disorder to undergo craniovertebral stabilization. METHODS: A 17-month-old boy presented with inability to support his head, decreasing muscle strength in all extremities, distended abdomen, and shortness of breath. The patient was found to have a dilated cardiomyopathy, hepatosplenomegaly, abnormal hepatobiliary function, corneal clouding, and a questionable tracheal anomaly. Genetic testing provided a diagnosis of Sly syndrome, mucopolysaccharidosis type VII. Magnetic resonance imaging revealed focal stenosis with significant spinal cord compression at the craniovertebral junction. Neurologic examination revealed normal muscle volume with strength 3/5 in all extremities and significant weakness in the neck muscles with instability at the craniovertebral junction. RESULTS: On a concerted preoperative medical clearance by pediatric intensive care, pediatric neuroanesthesia, pediatric cardiology, pediatric gastroenterology, and pediatric neurosurgery, the patient underwent occipital to C3 decompression and fusion with autogenous rib grafts. The patient was placed in a prefitted halo-vest after surgery and was neurologically intact. CONCLUSIONS: This case demonstrates the heterogeneity of cervical spine deformities among the mucopolysaccharidosis syndromes and confirms the propensity for deposition of glycosaminoglycans at the craniovertebral junction. Further studies should investigate the etiology for this propensity of glycosaminoglycan deposition at the craniovertebral junction. We think that this case demonstrates that, with appropriate preoperative planning, these patients can undergo successful posterior cervical arthrodesis despite their age or metabolic defects. PMID- 15129090 TI - Reversal of neurologic deficit in an adult gorilla with severe symptomatic lumbar stenosis. AB - STUDY DESIGN: A case report is presented. OBJECTIVE: To describe a case of severe symptomatic lumbar stenosis in an adult gorilla successfully treated with decompressive surgery. SUMMARY OF BACKGROUND DATA: Although spinal degenerative disease is thought to be less common in apes than in humans, gorillas and bonobos are known to have higher rates of disease than other African apes. METHODS: A 42 year-old female gorilla presented with a history of progressive lower extremity weakness. She was unable to ambulate normally or climb, pulling herself around with her arms. Her keeper did not believe she was incontinent. Physical examination revealed that ankle plantarflexion and dorsiflexion were absent on the right and weak on the left. Radiographs revealed disc space narrowing with osteophytes at the last mobile segment. Myelogram and computed tomography revealed severe spinal stenosis at the last two mobile segments. Under general anesthesia, a standard laminectomy and bilateral lateral recess decompression were performed. RESULTS: There were no anesthetic or wound complications. The gorilla was noted to have improved ankle strength bilaterally on postoperative day 1. On postoperative day 3, the animal began to walk and climb. At 20 months, the animal is back to her neurologic baseline. CONCLUSIONS: Symptomatic lumbar degenerative disease with neurologic compromise may be treated surgically, with excellent outcome, in the adult gorilla. Though this is an unusual situation, knowledge that this is a potentially reversible condition may benefit other gorillas in captivity. PMID- 15129091 TI - Eosinophilic granuloma of the atlas presenting as torticollis in a child. AB - STUDY DESIGN: This report describes a case of successful surgical treatment of eosinophilic granuloma of the atlas in a 3.5-year-old boy who presented with torticollis. OBJECTIVE: The purpose of this report was to illustrate the rare clinical presentation of eosinophilic granuloma in the atlas. SUMMARY OF BACKGROUND DATA: Eosinophilic granuloma, a benign solitary lesion that commonly affects children, has a variable clinical course. Although eosinophilic granuloma has been reported to occur in the cervical spine, there have been only five reported cases of eosinophilic granuloma affecting the atlas. The management of eosinophilic granuloma ranges from observation and immobilization of the cervical spine to surgical excision. METHODS: The tumor was located on the left lateral mass of the atlas. Biopsy and curettage were performed through an oblique incision through the posterior border of the sternocleidomastoid muscle. A histopathologic evaluation confirmed the diagnosis of eosinophilic granuloma. RESULTS: The patient tolerated the procedure with no complications, and his torticollis has completely resolved. CONCLUSION: Eosinophilic granuloma of the atlas initially presented in this patient as torticollis. Biopsy for confirmation of diagnosis and curettage for treatment provided a successful outcome in this patient. We suggest biopsy of the lesion and histopathologic evaluation to confirm the diagnosis in atypical cases. PMID- 15129093 TI - An analysis of the single-stage tympanoplasty with over-underlay grafting in tympanosclerosis. AB - OBJECTIVES: Our objective was to analyze the results of tympanosclerosis surgery using over-underlay tympanoplasty and to find out the effect of single-stage surgery on hearing results. MATERIALS AND METHODS: Forty-two patients who were operated on for tympanosclerosis between July 1998 and February 2002 were included in the study. These were one-stage tympanoplasties, because second-stage operations and revisions were not included. Operative records and audiograms of the patients were obtained. Of the patients, 48% had bilateral tympanosclerosis. In three (7.1%) of the patients, tympanosclerosis and cholesteatoma occurred concomitantly. The cog was present in eight (20%) of 40 mastoidectomies. Korner's septum was present in nine (22.5%) patients. The graft take rate was 95.2%. RESULTS: The pre- and postoperative air-bone gap values of the patients were not significantly different (p > 0.05). Thirty-three percent of the patients met the successful hearing criteria (air-bone gap <20 dB). However, 47% had an air-bone gap closure between 20 dB and 30 dB, whereas 20% had a gap of more than 30 dB. CONCLUSION: Single-stage surgery does not result in a satisfactory hearing improvement in most of the patients with tympanosclerosis. Only one third of the patients, most of whom had a mobile stapes, had satisfactory hearing results. Mobilization of a fixed stapes is not an effective option for hearing restoration in tympanosclerosis. Second-stage surgery for stapedectomy and placement of a prosthesis-like piston or total ossicular replacement prosthesis should be considered to obtain better hearing results in tympanosclerosis. PMID- 15129094 TI - Pathology of the ossicular chain: comparison between virtual endoscopy and 2D spiral CT-data. AB - OBJECTIVE: The purpose of this study was to assess the value of virtual endoscopy (VE) in the diagnosis of lesions of the ossicular chain and to compare virtual endoscopy and two-dimensional (2D) spiral computed tomography (CT) data. STUDY DESIGN: Retrospective study. SETTING: A university hospital. PATIENTS: Fifty eight patients with suspected ossicular chain lesions underwent a high-resolution CT of the temporal bone with both 2D data and VE before surgery. MAIN OUTCOME MEASURES: Two views were chosen for VE. The CT data obtained (2D, VE, and both 2D and VE) were compared with the lesions noted during surgery. RESULTS: In the diagnosis of dislocation of the ossicles or prostheses, VE seemed to be a better technique than 2D CT. Views chosen for the VE proved to be ineffective for diagnosing epitympanic fixations. VE was not adapted to the study of otosclerosis. We found it necessary to use the data provided by the addition to 2D CT data in the diagnosis of ossicular lysis and minor aplasia. Radiologic analysis of the lesions of the long process of the incus and the incudostapedial joint was improved by performing both 2D CT and VE. VE reconstruction of the stapes proved to be difficult, especially in cases of inflammation of the middle ear. CONCLUSION: This study demonstrates the value of VE in the diagnosis of dislocation of the ossicles and ossicular prostheses. VE was less effective in diagnosing other pathologies of the ossicular chain. PMID- 15129095 TI - Fibrin tissue adhesive and autologous concha cartilage for reconstruction of the posterior-superior canal wall of the chinchilla middle ear. AB - HYPOTHESIS: We conducted this study to prove that fibrin tissue adhesive (FTA) is safe, efficacious, biocompatible, and readily biodegradable with no deleterious side effects for fixation of a cartilage graft to bone along the chinchilla canal wall. METHODS: A posterior-superior canal defect was created in 12 chinchillas. The canal walls of six chinchillas were closed with autologous concha cartilage alone, whereas the canal wall of the remaining six animals were closed with cartilage in conjunction with fibrin tissue adhesive. RESULTS: Animals were killed 8 weeks postoperatively. Three of six cartilage grafts were displaced in the graft alone group, whereas all six grafts in the cartilage with FTA group healed without displacement. CONCLUSION: Fibrin tissue adhesive was found to be effective, biocompatible, biodegradable, and without any deleterious side effects for reconstruction of the superior-posterior canal wall of chinchillas. PMID- 15129096 TI - Malleus relocation in ossicular reconstruction: managing the anteriorly positioned malleus: results in a series of 268 cases. AB - OBJECTIVE: The objective of this study was to report an original method of malleus relocation allowing for better placement of both partial and total prostheses in ossicular reconstruction. STUDY DESIGN: We conducted a retrospective review of clinical and audiometric findings. SETTING: A tertiary referral center. MATERIALS AND METHODS: This is a study of 268 patients who underwent ossiculoplasty surgery from October 1997 to October 2000 for chronic otitis media, noninflammatory disease, and otosclerosis revision. Malleus relocation with total and partial ossicular replacement prostheses was used in all cases. Audiometric assessment included pre- and postoperative audiometric evaluation using conventional audiometry. Air-bone gap, bone-conduction thresholds, and air-conduction thresholds were measured. RESULTS: A postoperative air-bone gap closed to within 10 dB was achieved in 56% of cases. An air-bone gap smaller than 20 dB was obtained in 78% of cases. Postoperative improvement of air conduction thresholds superior to 20 dB was found in 41.5% of cases. The postoperative bone-conduction thresholds were unchanged in 98% of cases. One case of total postoperative sensorineural hearing loss was seen in this series (0.4%). Extrusion of the protheses was not observed in this series. Follow up ranged from 6 to 36 months (mean, 12.4 months). CONCLUSION: This study shows that malleus relocation is a safe and efficient technique for ossicular reconstruction. The ideal position of the relocated malleus allows easier and more stable placement of middle ear prostheses. PMID- 15129097 TI - Invasive middle ear cholesterol granuloma involving the basal turn of the cochlea with profound sensorineural hearing loss. AB - STUDY DESIGN: A retrospective surgical case review. SETTING: A tertiary-care, university teaching hospital. PATIENTS: The authors describe a unique case operated on for a middle ear cholesterol granuloma, which had invaded the cochlea and vestibule causing profound sensorineural deafness. INTERVENTIONS: Extended radical mastoidectomy and labyrinthectomy with musculofascial seal. RESULTS: Successful postoperative outcome with no recurrence seen after 2 years of follow up. CONCLUSION: To the authors' knowledge, this is the first reported case of a primary middle ear cholesterol granuloma with direct invasion into the cochlea. Such invasion of the otic capsule by cholesterol granulomas is rare and presents a diagnostic challenge to the attending otologist and radiologist. Salient points of the case history, pathogenesis, imaging studies, histopathology, and management are presented with a review of the current literature. PMID- 15129098 TI - Transfacial recess ossicular reconstruction: technique and early results. AB - OBJECTIVES: The objective of this study was to present the technique of transfacial recess ossicular chain reconstruction (TFROCR) for potential use in selected patients with cholesteatoma. STUDY DESIGN: The author conducted a retrospective case review of all candidates for TFROCR between August 1998 and March 2003. SETTING: A tertiary referral center. PATIENTS: At first-stage tympanomastoidectomy, 22 ears (21 patients; 8 children and 13 adults) with cholesteatoma and ossicular discontinuity were identified as candidates for TFROCR. Seven patients had undergone previous tympanomastoid surgery. INTERVENTION: The first stage included canal wall up mastoidectomy with resection of disease, wide opening of the facial recess, cartilage graft tympanoplasty, and placement of silicone elastomer in the middle ear. Approximately 6 months later, patients underwent a second-stage postauricular procedure. Endoscopes were used to inspect the middle ear through the facial recess. When possible, TFROCR was then performed without elevating a tympanomeatal flap. MAIN OUTCOME MEASURES: Variations in anatomy, disease control, and hearing results were studied. RESULTS: Of the 22 candidate ears, 17 successfully underwent TFROCR, whereas 5 required traditional second-stage procedures with canal incisions. There were no surgical complications. Early hearing results are promising with an average air bone gap of less than 20 dB. There have been no early failures from recurrent disease or prosthesis displacement. CONCLUSIONS: In carefully selected patients, TFROCR could be safe and effective for disease control and hearing restoration. It could provide for optimal prosthesis placement and almost immediate hearing improvement, avoiding the need for canal incisions, middle ear packing, and dry ear precautions. One must consider the potential risk of missing residual disease secondary to limited exposure. PMID- 15129099 TI - Middle temporal artery flap in mastoid surgery. AB - OBJECTIVE: The objective of this study was to describe the anatomy of the middle temporal artery (MTA) flap and its application in mastoid surgery. STUDY DESIGN: A description of the anatomy and surgical technique. CONCLUSION: The middle temporal flap is extremely useful in lining mastoid cavities, especially those in which poor healing is anticipated. It is available in most cases and is easily harvested and inset. It is particularly useful in revision cases, if present. PMID- 15129100 TI - Magnetic resonance imaging findings in the auditory pathway of patients with sudden deafness. AB - OBJECTIVES: The objectives of this study were to evaluate magnetic resonance imaging (MRI) findings of patients with sudden sensorineural hearing loss (SSNHL) and to grade the findings based on their clinical importance. STUDY DESIGN: A prospective clinical study. SETTING: A tertiary referral center (university hospital). PATIENTS: MRI findings of 82 consecutive patients with SSNHL fulfilling the inclusion criteria. MAIN OUTCOME MEASURES: We studied 1.0-T MR images that were analyzed by one experienced neuroradiologist. RESULTS: Of the six cases (7%) in which clearly hearing loss was obviously associated with the observed pathology, four patients had an acoustic neuroma in the internal auditory canal or cerebellopontine angle, one patient had changes at the level of pons, and one patient had an obliterated internal carotid artery. Of the six other patients (7%) in which MRI revealed changes that suggest a possible etiology to hearing loss, two patients showed a demyelinating process and four patients showed blood vessel abnormalities such as caroticocavernous fistula, abnormally locating vertebral or basilar artery, and a venous angioma. CONCLUSIONS: Enhanced MR imaging seems to be a useful examination in patients with SSNHL. The aim should not be only to exclude specific retrocochlear etiologies, but by appropriate techniques, MRI could reveal both peripheral and central abnormalities. PMID- 15129101 TI - Autoimmune mouse antibodies recognize multiple antigens proposed in human immune mediated hearing loss. AB - HYPOTHESIS: Autoimmune diseased mice with hearing loss will have autoantibodies against the various cochlear antigens proposed in clinical autoimmune inner ear disease. BACKGROUND: Serum antibodies of patients with hearing loss recognize several proteins that are proposed as possible antigenic targets in the ear. This often leads to a clinical diagnosis of autoimmune inner ear disease, although it is not clear how these antibodies cause inner ear disease. Therefore, to better understand the relationship of autoantibodies and ear disease, an examination was made of serum autoantibodies in the MRL/MpJ-Fas(lpr) autoimmune mouse with hearing loss. Similar antibody patterns in the mouse would provide an animal model in which to investigate potential autoimmune mechanisms of this clinical ear disorder. METHODS: Sera from MRL/MpJ-Fas(lpr) autoimmune mice and normal C3H mice were tested by the enzyme-linked immunosorbent assay technique for reactivity against various reported cochlear antigens: heat shock protein 70 (bovine, human, bacterial), laminin, heparan sulfate proteoglycan, cardiolipin, and collagen types II and IV. RESULTS: The autoimmune mouse sera showed significantly greater antibody reactivity against all of the antigens when compared with normal mouse sera. CONCLUSIONS: Serum antibodies from autoimmune mice recognized several putative autoantigens reported for patients with hearing loss, suggesting that comparable antigen-antibody mechanisms might be operating. However, the recognition of multiple antigens did not identify any one as being the specific target in autoimmune hearing loss. The correlation of antibodies in the MRL/MpJ-Fas(lpr) autoimmune mouse and human studies indicates this animal model should aid further investigations into potential cochlear antigens in autoimmune hearing loss. PMID- 15129102 TI - Histologic evaluation of the tissue seal and biologic response around cochlear implant electrodes in the human. AB - HYPOTHESIS: Histopathologic study of the tissue seal and biologic response around cochlear implant electrodes in patients who had received a cochlear implant during life could provide clues concerning the pathogenesis of meningitis after cochlear implantation. BACKGROUND: Bacterial meningitis has been reported as an infrequent complication of cochlear implantation using a variety of electrode designs. The cause of meningitis in cochlear implant recipients has not been firmly established. In an analogous surgical situation, namely stapedectomy, delayed meningitis could occur as a complication of ipsilateral acute suppurative otitis media in which there was open communication between the middle ear and perilymph. METHODS: Twenty-one temporal bones from 20 individuals who had undergone cochlear implantation during life were studied by light microscopy. All sections passing through the cochleostomy site and electrode track were examined to evaluate the tissue seal at the cochleostomy, the presence or absence of an extracochlear electrode sheath, and finally, to seek evidence of a cellular inflammatory response near the electrode. These data were compared with clinical data, including electrode system used, the number of years between implantation and death, type of tissue used at surgery, and the age and sex of the patients. RESULTS: The 21 specimens included cases implanted with the Symbion Ineraid, Cochlear Corporation Nucleus 22-channel, Cochlear Corporation Nucleus 24-channel, a Cochlear Corporation Nucleus single channel, and Advanced Bionics Clarion C1 devices. At the cochleostomy site, and just within the cochlea, there was a robust fibrous and bony tissue response in all 21 ears and in most cases, there was a fibrous sheath surrounding the electrode in the middle ear. No recognizable open communication or potential communication between the middle ear and the inner ear was seen in any of the 21 ears. An inflammatory cellular response, including mononuclear leukocytes, histiocytes, and foreign body giant cells, were present in 12 of the 21 temporal bones (57%) and was most intense at the cochleostomy site. No statistically significant relationship was found between the presence or absence of inflammatory cells and the type of tissue graft used at surgery. CONCLUSIONS: The histologic evidence presented in this study does not support open communication between the middle and the inner ear as part of the pathogenesis of bacterial meningitis as a late complication after cochlear implantation. Rather, the finding of a cellular inflammatory response in 12 of 21 temporal bones suggests that late hematogenous contamination and colonization of the implant is a much more likely pathogenic mechanism. This putative mechanism has implications for possible strategies to prevent meningitis after cochlear implantation. PMID- 15129103 TI - Cochlear implantation between 5 and 20 months of age: the onset of babbling and the audiologic outcome. AB - OBJECTIVE: The objective of this study was to investigate the onset of prelexical babbling and the audiologic outcome of 10 deaf children who received a cochlear implant (CI) before the age of 20 months. STUDY DESIGN: A prospective longitudinal observation and analysis. PATIENTS: Ten congenitally deaf infants implanted at an age between 6 and 18 months. INTERVENTION: All children received a Nucleus-24 multichannel cochlear implant. MAIN OUTCOME MEASURES: 1) The onset of babbling defined as a) the first appearance of multiple articulatory movements and b) a canonical babbling ratio of.2 or higher; 2) the babbling spurt defined as a sudden increase of babbled utterances; 3) the audiologic outcome defined by the CAP score (Categories of Auditory Performance) and the results of the A[S]E (Auditory Speech Sound Evaluation). RESULTS: All children started babbling after a short interval of 1 to 4 months after activation of the device so that the onset of babbling in the youngest subjects occurred at a chronologic age comparable to that of normally hearing infants. The outcomes of the different babbling measures correlated significantly with the age of implantation: the earlier the implantation, the closer the results approached the outcomes of normally hearing infants. The children implanted in their first year of life showed a normal CAP development as early as 3 months after implantation. All CI children were able to discriminate phoneme pairs of the A[S]E immediately after the fitting of the device. CONCLUSIONS: The earlier the implantation took place, the smaller the delay was in comparison with normally hearing children with regard to the onset of prelexical babbling and with regard to auditory performance as measured by CAP. PMID- 15129104 TI - Interobserver agreement of coiling of Med-El cochlear implant: plain x-ray studies. AB - OBJECTIVE: The objective of this study was to describe the interobserver agreement of otologists in judging the extent of cochlear electrode insertion as depicted in plain radiographs done in various projections relative to that of Stenvers (45 degrees from midline). STUDY DESIGN: Ten experienced U.S. MED-EL cochlear implant surgeons, unaware of the image history, judged depths of electrode coiling in 12 images. METHODS: In a bequeathed cranial base specimen, a standard MED-EL Combi 40+ electrode was inserted 3 mm less than fully and stabilized. With a custom cephalostat, the specimen was positioned in the Frankfort horizontal plane. Plain radiographs were made in six projections in 15 degrees increments, from posterior-anterior to near lateral. After the electrode array was withdrawn 5 mm and stabilized, six additional radiographs were similarly taken. RESULTS: Greater variability of assessed coiling was associated with projections other than Stenvers. The coefficient of variation for the Stenvers projection was significantly less than half that of the anterior posterior and the other projections studied. CONCLUSION: In plain radiographic assessment of the degree of coiling of the MED-EL electrode, interobserver agreement was best for the Stenvers view. PMID- 15129105 TI - Meningitis in cochlear implant recipients: the North American experience. AB - INTRODUCTION: Until recently, postimplant meningitis was infrequently reported and felt to be uncommon. However, in the spring of 2002, there was a sudden increase in the number of reported cases of postimplantation meningitis in both Europe and North America. OBJECTIVE: Because complications of surgery often tend to be underreported, we decided to survey all cochlear implant centers in North America to determine the true incidence of postimplant meningitis and to learn more about the demographics and risk factors. STUDY DESIGN: We conducted a prospective study. A survey instrument was designed asking surgeons the number of implants performed and whether they had seen any cases of meningitis after implantation. If the answer was affirmative, they were asked to respond to a 20 point questionnaire. This instrument was sent to all 401 cochlear implant centers in North America. SETTING: Tertiary care referral centers. PATIENTS: We studied all patients having received cochlear implants in North America. MAIN OUTCOME MEASURES: Number of cases of postimplant meningitis, age of patients, device used, cochlear and temporal bone abnormalities, treatment, and outcomes. RESULTS: Meningitis is more common than previously thought. Risk factors included young age, cochlear dysplasia, temporal bone abnormalities, and the use of a two-part electrode system. This survey led to the involvement by the U.S. Food and Drug Administration and the Centers for Disease Control and Prevention in a much more intensive analysis of a subset of the cases. CONCLUSIONS: Postimplant meningitis is related to patient, surgical, and device factors. By being aware of the risk factors involved, adhering to sound surgical principles such as packing of the cochleostomy with soft tissue, appropriately vaccinating patients, and eliminating any potentially traumatic electrode arrays, the incidence of meningitis should be significantly diminished. PMID- 15129106 TI - The facial nerve canal: an important cochlear conduction path revealed by Clarion electrical field imaging. AB - HYPOTHESIS: Electrical properties of the implanted scala tympani could be accurately modeled by means of a simple resistive ladder network model. The subject-specific model parameters can be obtained from electrical field imaging (EFI) recordings. It is a powerful tool for analysis of the cochlear current spread. BACKGROUND: In EFI mode, the telemetry systems of contemporary cochlear implants can measure the intracochlear potential distribution. At present, the clinical use of EFI is typically limited to checking the implant's proper functioning. METHODS: Accurate EFI measurements and estimation algorithms have been developed to fit a small, yet physically relevant electrical model of the conductivity of the intracochlear structures. RESULTS: The model can attain up to 95% agreement with in vivo EFI data. A first discovery is that in a majority of the tested subjects, a substantial fraction of the monopolar current leaves the scala along the facial nerve canal. The role of the facial nerve canal has been confirmed by a temporal bone study and a high-resolution computed tomography (HRCT) scan in two of the implanted subjects. CONCLUSIONS: The clinical use of EFI is not limited to checking the implant's status. For the Clarion II implant, a purely resistive model is able to match in vivo EFI recordings. The model indicates that the facial nerve canal is an important conduction path to the reference electrode. EFI can provide clinically relevant information, especially in problematic cases of cochlear malformations, postoperative fibrosis/ossification, implanted otosclerotic cochleae, postoperative facial nerve stimulation, increased stimulation thresholds, and so on. PMID- 15129107 TI - Degree of modiolar coiling, electrical thresholds, and speech perception after cochlear implantation. AB - OBJECTIVE: Perimodiolar electrode arrays were developed to improve stimulation of specific neural populations and to decrease power consumption. Postoperative radiographs suggest that some arrays are more tightly coiled than others. The purpose of this study was to evaluate whether the degree of modiolar coil correlates with electrical threshold and/or performance measures postimplantation with the Nucleus CI24RCS (Contour) device. STUDY DESIGN: Retrospective review. SETTING: Cochlear implant center and a tertiary care hospital. PATIENTS: Twenty eight adult cochlear implant recipients with normal cochlear anatomy who underwent implantation with the Nucleus C124RCS perimodiolar electrode and are at least 1 year postimplantation. INTERVENTIONS: Therapeutic. MAIN OUTCOME MEASURES: Radiologic degree of perimodiolar electrode placement (determined by a computer algorithmic analysis) compared with electrical thresholds and standard speech perception outcome measures at 1 year postcochlear implantation. RESULTS: No significant correlations were found between the degree of modiolar coiling of the electrode array and electrical thresholds and speech perception outcome measures at 1 year postcochlear implantation. CONCLUSIONS: The degree of coiling of a modiolar hugging electrode array was not directly correlated with the level of electrical thresholds or postoperative speech perception outcome measures. Appearance of coil tightness on postoperative radiographs could reflect either differences in array placement or intrinsic variations in cochlear anatomy, and variations in speech perception performance can be influenced by other factors, including length of deafness. PMID- 15129108 TI - Prospective evaluation of intraoperative cochlear implant radiographs. AB - OBJECTIVE: To investigate the clinical utility of intraoperative plain radiographs in cochlear implant surgery. STUDY DESIGN: Prospective. SETTING: Tertiary referral center. PATIENTS: Eighty consecutive adult and pediatric cochlear implant operations at a facility capable of intraoperative radiographs were evaluated over 12 months. INTERVENTIONS: Intraoperative plain radiographs to assess implant location and surgeon questionnaire. MAIN OUTCOME MEASURES: Plain radiographic image interpretation and surgeon questionnaire. RESULTS: In one revision surgery, the intraoperative plain radiograph was useful for confirming the cochleostomy site. In the remaining 79 operations, no changes in the electrode arrays were made on the basis of the information provided by intra operative plain radiographs. CONCLUSIONS: In this cohort, intraoperative plain radiographs were not useful for uncomplicated implant operations; however, they may be useful for complicated operations. These results may have implications for surgical cost and patient radiation exposure. PMID- 15129109 TI - Cochlear implant outcomes in the elderly. AB - OBJECTIVE: This study aimed to review cochlear implantation with respect to surgical and auditory outcomes in subjects aged 70 years and older. STUDY DESIGN: Retrospective chart review. SETTING: Tertiary referral centers. PATIENTS: Sixty five patients aged 70 years or older at the time of implantation were compared to a group of patients aged <70 years. INTERVENTION: Patients underwent multichannel cochlear implantation with either the Clarion or Nucleus device. MAIN OUTCOME MEASURE: Presence or absence of surgical complications and auditory performance with open-set word and sentence recognition testing. RESULTS: In patients implanted at age 70 or older, significant improvement in speech understanding was demonstrated in performance scores using Consonant Nucleus Consonant words, Central Institute for the Deaf sentences, and Hearing in Noise Test sentences at 3, 6, and 12 months when compared to preimplantation scores. However, their performance was slightly poorer when compared to a control group of patients <70 years of age in the same measures at 3, 6, and 12 months. CONCLUSIONS: The elderly population showed significant improvement in auditory performance tests following cochlear implantation compared to their preimplantation scores but performed less well than younger patients. PMID- 15129110 TI - Evidence of subtle auditory deficit in a group of patients recovered from bacterial meningitis. AB - OBJECTIVES: Sensorineural hearing loss of greater than 30 dB hearing loss occurs in up to 30% of patients after acute bacterial meningitis. This study investigated whether postbacterial meningitic patients with no apparent clinical sensorineural hearing loss had any evidence of more subtle subclinical cochlear deficit. DESIGN: Prospective case-controlled clinical trial. SETTING: Departments of Otolaryngology in Leicester and Nottingham, England, UK. PATIENTS: Fifty-eight controls and 20 postbacterial meningitic patients aged between 18 and 38 years were screened by a questionnaire and tympanometry to exclude hearing loss attributable to other causes. All participants fell below the 90th percentile pure-tone audiometry threshold of the Lutman and Davis UK data sets. MAIN OUTCOME MEASURES: In both ears, standard (0.25-8 kHz) pure-tone audiometry, high frequency pure-tone audiometry (10-16 kHz), and distortion product otoacoustic emissions at 2, 4, and 6 kHz were measured. RESULTS: Mean thresholds over the range of standard pure-tone audiometry (analyzed independently) for the postbacterial meningitic patients were significantly elevated at most frequencies (p < 0.05-p < 0.001) between 4 and 7 dB in both ears above control group values. There was no evidence of significant high-frequency threshold elevation (10-16 kHz). The mean iso-distortion product values at 2, 4, and 6 kHz were elevated in both ears in the meningitis group; significantly so (p < 0.05-p < 0.01) at all three frequencies in the right ear and at 4 kHz in the left. CONCLUSIONS: Postbacterial meningitic patients with hearing below the 90th percentile range had a slight but significant subclinical threshold elevation over the standard pure-tone audiometry. This may reflect a real effect of the infection at the level of the cochlea, or it may be attributable to a mild residual cognitive defect. The moderate increases in iso-distortion product values are more likely to be real and reflect an effect on outer hair cell function in response to lower stimulus intensities. PMID- 15129111 TI - Congenital malformation of the inner ear and pediatric cochlear implantation. AB - OBJECTIVES: To study the surgical aspects and performance outcome of cochlear implantation in children with malformed inner ears. STUDY DESIGN: Clinical and audiometric evaluation in 13 patients. METHODS: Patient data concerning surgery, postoperative follow-up, and pre- and postimplantation audiometry were obtained from the cochlear implant center's database and evaluated. A review of the literature has been included. SETTING: Tertiary referral center. PATIENTS: The patients had a variety of inner ear malformations and profound hearing loss. One patient with recurrent meningitis had a severe cochlear malformation (common cavity). RESULTS: Major complications did not occur. In one patient with an abnormal position of the cochlea and concurring middle ear disease, it was difficult to find the scala tympani during surgery. A cerebrospinal fluid gusher was encountered in two patients and an aberrant facial nerve in another, which did not lead to any complications. The patients with mild cochlear malformation such as an incomplete partition demonstrated a good performance in speech perception tests. Even the child with the common cavity deformity had some open set speech perception 1 year after implantation. CONCLUSIONS: Viewing the patients from this study and patients from a review of the literature concerning cochlear implantation in children with malformed inner ears including severe cochlear malformations, the occurrence of an aberrant facial nerve was 17%, which increases to 27% if one reviews the surgical findings in children with severe malformed cochleae such as a common cavity or a severe cochlear hypoplasia. In the latter patients, results in speech perception vary. Although the result of cochlear implantation may be promising, as in our patient with a common cavity, during preoperative counseling the child's parents must be informed that the result is uncertain. PMID- 15129112 TI - Neonatal middle ear effusion predicts chronic otitis media with effusion. AB - HYPOTHESIS AND AIMS: The specific aims of the research are to determine whether newborn ears with persistent middle ear effusion at age 30 to 48 hours are more likely to develop chronic otitis media with effusion over the first year of life when compared with ears without persistent middle ear effusion. The hypothesis is that neonates with middle ear effusion persisting to 30 to 48 hours are more likely to develop chronic otitis media with effusion. STUDY DESIGN: Prospective, case-control design. Loupe-magnified pneumatic otoscopy performed at the time of newborn hearing screening determined presence or absence of effusion. Infants enrolled in the study returned for outpatient examinations. SETTING: University medical center well-baby nursery and out-patient audiology clinic. SUBJECTS: From 454 neonates, 14 experimental subjects with neonatal middle ear effusions and 15 control subjects free of neonatal effusion were recruited for the study and followed-up for 1 year. INTERVENTIONS: Outpatient study tests included transient evoked otoacoustic emissions, tympanometry, pneumatic otoscopy, and visual reinforcement audiometry (starting at age 6 months), at 3, 6, 9, and 12 months of age. Experimental (neonatal effusion) infants were followed-up starting at age 1 month. Infants found at any follow-up examination to have effusion on otoscopy were followed-up and tested 1 month later. MAIN OUTCOME MEASURES: Chronic otitis media with effusion defined as hypomobile or immobile tympanic membrane on pneumatic otoscopy in one or both ears for three consecutive monthly examinations. Hearing loss defined as greater than 25-dB hearing loss visual reinforcement audiometry thresholds. RESULTS: Eight experimental infants (58%) and three control infants (20%) developed chronic otitis media with effusion (p < 0.04). The average number of effusions was 1.27 for control and 4.14 for experimental infants (average number of effusions for each group at 3-, 6-, 9-, and 12-month visits). Warbled tone and speech visual reinforcement audiometry thresholds averaged 3 dB worse in the experimental group, but these differences were not statistically significant. For the control group, mean visual reinforcement audiometry thresholds never exceeded 25 dB hearing loss. For the experimental group, mean visual reinforcement audiometry thresholds exceeded 25 dB hearing loss at 1,000, 2,000, and 4,000 Hz at 9 months. CONCLUSIONS: A majority of infants with persistent neonatal middle ear effusion found by pneumatic otoscopy at 30 to 48 hours will develop chronic otitis media with effusion during the first year of life. However, chronic otitis media with effusion is common in all infants (20% of controls), a time during which infants are examined and tested frequently. PMID- 15129113 TI - Clinical, experimental, and theoretical investigations of the effect of superior semicircular canal dehiscence on hearing mechanisms. AB - HYPOTHESIS: A superior semicircular canal dehiscence affects hearing by introducing a third window into the inner ear that 1) lowers cochlear input impedance, 2) shunts air-conducted sound away from the cochlea resulting in conductive hearing loss, and 3) improves bone-conduction thresholds by increasing the difference in impedance between the vestibule and the round window. BACKGROUND: Superior semicircular canal dehiscence has been linked to a "conductive" hearing loss characterized by a decrease in the sensitivity to air conducted sound and hypersensitivity to bone-conducted sound. METHODS: Four investigations were performed: 1) laser-Doppler vibrometer measurements of sound induced umbo velocity in patients with computed tomographic scan-confirmed superior semicircular canal dehiscence; 2) laser-Doppler vibrometry of sound induced motions of the vestibular lymph (either perilymph or endolymph) exposed in a chinchilla model of superior semicircular canal dehiscence; 3) studies in chinchillas of the effect of superior semicircular canal dehiscence on the cochlea's sensitivity to bone-conducted sounds; and 4) anatomically based theoretical analyses of sound flow through the human cochlea and semicircular canals. RESULTS: The low-frequency umbo velocity in superior semicircular canal dehiscence patients without previous middle ear surgery ranged from normal through high normal. This tendency toward hypermobility suggests a decrease in cochlear impedance. Measurements of sound-induced velocity of the lymph within a superior semicircular canal dehiscence in chinchillas demonstrated sound flow through the dehiscence. Measurements of the cochlear potential demonstrated a superior semicircular canal dehiscence-induced increase in response to bone conducted sound in eight of nine chinchillas. An anatomically based model of the human ear predicts changes in auditory sensitivity similar to audiometric changes in superior semicircular canal dehiscence. CONCLUSION: The results suggest that superior semicircular canal dehiscence can affect hearing function by introducing a third window into the inner ear. PMID- 15129114 TI - Vestibular evoked myogenic potentials show altered tuning in patients with Meniere's disease. AB - OBJECTIVE: Acoustic stimulation of the saccule gives rise to a vestibulocollic reflex, the output of which can be measured in the neck as inhibition of activity in the ipsilateral sternocleidomastoid muscle. This vestibular evoked myogenic potential has been promoted as a means of assessing integrity of saccular function. In this study, we test the hypothesis that the cochleosaccular hydrops of Meniere's syndrome leads to alterations in saccular motion that change the dynamics of the vestibular evoked myogenic potential. STUDY DESIGN: Prospective cohort study. SETTING: Large specialty hospital, department of otolaryngology. SUBJECTS: Fourteen normal adult volunteers and 34 consecutive consenting adult patients with unilateral Meniere's disease by American Academy of Otolaryngology Head and Neck Surgery diagnostic criteria. INTERVENTIONS: All subjects underwent vestibular evoked myogenic potential testing using ipsilateral broadband click and short tone-burst stimuli at 250, 500, 1,000, 2,000, and 4,000 Hz. MAIN OUTCOME MEASURES: Threshold, amplitude, and latency of vestibular evoked myogenic potential responses in normal and Meniere's affected and unaffected ears. RESULTS: Vestibular evoked myogenic potential was present in all ears tested. Normal subjects show a frequency-dependent vestibular evoked myogenic potential threshold, with best response ("frequency tuning") at 500 Hz. Compared with normal subjects and unaffected ears of Meniere's subjects, affected Meniere's ears had significantly increased vestibular evoked myogenic potential thresholds. Affected Meniere's ears showed threshold shifts at all frequencies and there was less tuning apparent at 500 Hz. Unaffected ears of Meniere's subjects also showed significantly elevated vestibular evoked myogenic potential thresholds compared with normal subjects. Analyses of vestibular evoked myogenic potential thresholds for effects of age, hearing loss, and audiometric configuration showed no significant differences. CONCLUSIONS: Meniere's ears display alterations in vestibular evoked myogenic potential threshold and tuning, supporting our hypothesis of altered saccular motion mechanics arising from hydropic distention. Unaffected ears of unilateral Meniere's subjects show similar changes, though to a lesser degree. This finding may be because of occult saccular hydrops in the asymptomatic ear or binaural interactions in the vestibular evoked myogenic potential otolith-cervical reflex arc. PMID- 15129115 TI - Evaluating quality of life after endolymphatic sac surgery: The Meniere's Disease Outcomes Questionnaire. AB - OBJECTIVE: To develop a disease-specific instrument to measure the quality of life in patients with Meniere's disease and to assess quality-of-life outcomes after endolymphatic sac decompression. STUDY DESIGN: Retrospective survey. PATIENTS: Patients with Meniere's disease who underwent endolymphatic sac decompression from June 1996 to June 2001, after failing a course of medical management. Two hundred fifteen potential subjects were identified; completed questionnaires were returned by 159 patients. MAIN OUTCOME MEASURES: The Meniere's Disease Outcomes Questionnaire was developed, and consists of questions in three domains that determine quality of life: physical, emotional, and social well-being. The Meniere's Disease Outcomes Questionnaire consisted of 18 multiple choice questions that were paired for pre- and postoperative conditions, and one global quality-of-life question. The preoperative quality-of-life score (total score for preoperative items) was compared with the postoperative quality-of-life score. The main outcomes measure was the change in quality-of-life score. RESULTS: Overall, the mean change in quality-of-life score was +25.6 points (range, -34 to 83) (p < 0.001). The change in Meniere's Disease Outcomes Questionnaire quality-of-life score was highly correlated with the change in the global question score (p < 0.01). Quality of life was improved in 87% of respondents, unchanged in 3% of patients, and poorer in 9% of patients after endolymphatic sac decompression. CONCLUSIONS: The Meniere's Disease Outcomes Questionnaire is a new disease-specific quality-of-life tool that is a valid measure of quality of life in patients with Meniere's disease, and is responsive to measuring change in quality of life after treatment. Significant improvement in quality of life was reported by 87% of patients after endolymphatic sac decompression. PMID- 15129116 TI - Acoustic responses of vestibular afferents in a model of superior canal dehiscence. AB - HYPOTHESIS: Afferents innervating the superior semicircular canal are rendered especially sensitive to acoustic stimulation when there is a dehiscence of the superior canal. Other vestibular end organs are also more sensitive to acoustic stimulation. BACKGROUND: Dehiscence of the superior semicircular canal is associated with vertigo and nystagmus caused by loud sounds (Tullio phenomenon) or changes in middle ear or intracranial pressures. The mechanisms by which acoustic stimuli act on the vestibular end organs are unclear. The nystagmus caused by acoustic stimuli generally aligns with the affected superior canal. METHODS: Responses to acoustic stimuli in the superior vestibular nerves of anesthetized chinchillas were recorded before and after fenestration of the superior canal. RESULTS: Two acoustic response patterns were seen: rapid phase locking and slow tonic changes in firing rate. Phasic responses principally occurred in irregular afferents and tonic responses in regular afferents. Afferents from all of the vestibular end organs encountered could respond to acoustic stimuli, even before fenestration. However, fenestration lowered the thresholds for acoustic stimulation in superior canal afferents with phasic responses and increased the magnitude of tonic responses. CONCLUSIONS: Superior canal dehiscence may render the irregular afferents innervating the superior canal particularly sensitive to loud sounds. Rapid phase-locking responses may explain the short latency of nystagmus seen in patients with superior canal dehiscence syndrome. The mechanisms by which acoustic stimuli activate the vestibular end organs may differ from the damped endolymph motion associated with head acceleration. PMID- 15129117 TI - Benign paroxysmal positional nystagmus in hospitalized subjects receiving ototoxic medications. AB - OBJECTIVE: To investigate the occurrence of benign paroxysmal positional nystagmus in subjects undergoing treatment with potentially ototoxic medications. STUDY DESIGN: Prospective and retrospective record reviews. SETTING: Tertiary referral neurotology clinic; clinical research and technology center. SUBJECTS: Ninety-nine hospitalized subjects undergoing treatment of infectious disease or carcinoma with potentially ototoxic medications. INTERVENTIONS: Records review, tests of vestibular function. MAIN OUTCOME MEASURE: Results of Hallpike positional tests for benign paroxysmal positional nystagmus (electro oculography). RESULTS: Forty-one (41%) of 99 subjects were female and 58 (59%) were male. Age range was 15 to 73 years (mean, 47 years). Forty-nine (50%) of 99 subjects had an unequivocally positive Hallpike test for benign paroxysmal positional nystagmus in one or both ears. The occurrence of benign paroxysmal positional nystagmus in the Hallpike-positive population was distributed equally across age decades. Of the 49 subjects with benign paroxysmal positional nystagmus, 22 (44%) were female and 27 (56%) were male. CONCLUSIONS: Benign paroxysmal positional nystagmus is the most common cause of vertigo in the general population, including subjects receiving ototoxic drugs. Complaints of vertigo in subjects receiving ototoxic drugs therefore may or may not indicate onset of ototoxicity. Occurrence of benign paroxysmal positional nystagmus in subjects receiving ototoxic drugs was independent of gender or age. The high occurrence rate of benign paroxysmal positional nystagmus in subjects receiving potentially ototoxic medications is consistent with the observation that benign paroxysmal positional nystagmus occurs in combination with many pathologic conditions. Benign paroxysmal positional nystagmus presenting in subjects receiving ototoxic drugs may complicate the clinical identification of ototoxicity and obfuscate clinical decision-making processes. PMID- 15129118 TI - Co-expression of transforming growth factor-beta1 and glial cell line-derived neurotrophic factor in vestibular schwannoma. AB - HYPOTHESIS: Transforming growth factor-beta1, glial cell line-derived neurotrophic factor, and their receptors are expressed in vestibular schwannoma, and the expression data correlate with the proliferation activity (Ki-67 labeling index) and the clinical growth rate of vestibular schwannoma tissue. BACKGROUND: Glial cell line-derived neurotrophic factor is a potent growth factor for the central and peripheral nervous system. Recent results demonstrate that glial cell line-derived neurotrophic factor requires transforming growth factor-beta to exert its trophic effect on neural tissue. A functional role, including that in Schwann cell proliferation, is discussed for both transforming growth factor beta1 and glial cell line-derived neurotrophic factor. METHODS: Immunohistochemical analysis for transforming growth factor-beta1 and glial cell line-derived neurotrophic factor and their receptors TbetaR II, GFRalpha-1, and Ret was performed on formalin-fixed, paraffin-embedded archival surgical specimens. The Ki-67 labeling index (mean Ki-67 labeling index and highest Ki-67 labeling index for Antoni Type A and Type B regions) and the clinical growth rate of vestibular schwannoma were determined and correlated with the expression patterns of the examined neurotrophic factors and their receptors. RESULTS: Results demonstrate co-expression of transforming growth factor-beta1 and glial cell line-derived neurotrophic factor with higher levels in Antoni Type A than in Antoni Type B regions. Ninety-five percent of vestibular schwannomas exhibited transforming growth factor-beta1 immunoreactivity, and glial cell line-derived neurotrophic factor expression was found in 100% of vestibular schwannoma specimens. Fifty percent of vestibular schwannoma displayed TbetaR II immunostaining, 100% showed positive reactions for GFRalpha-1, and 86% showed positive reactions for Ret. Statistical analysis revealed no significant correlation in neurotrophin expression related to sex, age, tumor size, clinical growth rate, or Ki-67-labeling indices. CONCLUSIONS: Expression of transforming growth factor-beta1 and glial cell line-derived neurotrophic factor may suggest a biological role for both growth factors in vestibular schwannomas. Trophic transforming growth factor-beta/glial cell line-derived neurotrophic factor synergism seems possible and is underscored by co-expression of both neurotrophic factors and their receptors. PMID- 15129119 TI - Postirradiation vertigo in nasopharyngeal carcinoma survivors. AB - OBJECTIVE: Vertigo rarely manifested as an initial symptom of nasopharyngeal carcinoma or an early symptom after irradiation; however, increasing numbers of long-term nasopharyngeal carcinoma survivors experienced it. The purpose of this study was to investigate the causes of vertigo in irradiated nasopharyngeal carcinoma survivors. SETTING: University hospital. PATIENTS: From January 1992 to December 2001, a total of 113 nasopharyngeal carcinoma patients (67 men and 46 women) with postirradiation vertigo consecutively visited our vertigo clinic. The mean interval from completion of irradiation to the occurrence of vertigo was 10 years. Each patient underwent otoscopic examination and a battery of audiovestibular function tests. Then, correlation between the vertigo and the radiation effect was explored. RESULTS: Postirradiation vertigo was mainly attributable to peripheral labyrinthine disorder (69%), followed by central vestibular lesions (31%). The vertiginous and associated symptoms including severity, nausea/vomiting, oscillopsia, or imbalance in cases of peripheral labyrinthine disorder were milder than those in central vestibular lesion. Meanwhile, the former had less life impact and better response to therapy compared with the latter. The mean radiation dosage in both groups was 73 +/- 6 Gy and 74 +/- 5 Gy, respectively, without a significant difference. Of these 113 patients, 85 patients (75%) had radiation otitis media in one or both ears, and 28 patients disclosed bilateral intact eardrums. The prevalence of radiation otitis media in patients with postirradiated vertigo was 53% versus 73% for those with radiation dosage less than or more than 71 Gy, respectively, exhibiting a significant difference. However, the prevalence of radiation otitis media is unrelated to radiation interval. In comparison with absent caloric responses in radiation otitis media ears, 32% versus 57% in those of less than or more than 71 Gy, respectively, indicates a significant difference. CONCLUSIONS: Vertigo is a late complication in irradiated nasopharyngeal carcinoma survivors, which is mainly caused by the sequela of radiation otitis media. Because the latter is correlated with the radiation dosage, 70 Gy is recommended as the maximum dosage for nasopharyngeal carcinoma. Eradicating radiation otitis media in nasopharyngeal carcinoma survivors may subsequently prevent the postirradiation vertigo. PMID- 15129120 TI - Change in hearing handicap after translabyrinthine vestibular schwannoma excision. AB - OBJECTIVE: To evaluate the change in hearing handicap after translabyrinthine vestibular schwannoma excision. STUDY DESIGN: Prospective administration of the Hearing Handicap Inventory preoperatively and at 3 and 12 months postoperatively. SETTING: A tertiary referral neurootology clinic. PATIENTS: A total of 119 consecutive patients who had vestibular schwannomas excised between May 1998 and July 2002 and who had completed Hearing Handicap Inventories preoperatively and at 3 and 12 months postoperatively. INTERVENTIONS: Translabyrinthine excision of a unilateral sporadic vestibular schwannoma; pre- and postoperative hearing therapy and rehabilitation. MAIN OUTCOME MEASURES: Hearing Handicap Inventory scores. RESULTS: Sixty-one percent of patients showed no change in their hearing handicap between preoperative and 3-month postoperative measures. Fifty-eight percent showed no change between preoperative and 12-month postoperative measures. Twenty-five percent of patients had a worsening in their hearing handicap when comparing preoperative and 12-month postoperative scores, and these patients were those with a better hearing class preoperatively. Seventeen percent of patients showed an improvement in their hearing handicap over this same time period, and the reasons for this remain unclear. Neither age nor tumor size had an effect on change in hearing handicap. CONCLUSIONS: The findings of the current study can be used during preoperative patient counseling. In particular, the clinician is now able to take an informed and positive stance about the hearing handicap to be expected postoperatively. PMID- 15129121 TI - Perioperative complications in acoustic neuroma (vestibular schwannoma) surgery. AB - OBJECTIVE: Retrospective study and review of the complications other than those related to the facial nerve and hearing, encountered in acoustic neuroma surgery. Also, an evaluation of hospital stay and its relation with various factors. STUDY DESIGN: Retrospective case review. SETTING: Tertiary neurotologic and skull base referral center. PATIENTS: A series of 707 patients who underwent surgical removal of acoustic neuroma from April 1987 to December 2001. INTERVENTIONS: The surgical approaches used were the enlarged translabyrinthine approach, the enlarged middle fossa approach, and the retrosigmoid approach. In a small number of cases, the operations were performed through other approaches. MAIN OUTCOME MEASURES: The duration of hospital stay and appearance of complications in the perioperative period along with their management. Results related to the facial nerve and hearing were not considered in this study. RESULTS: The most frequent complication was abdominal subcutaneous hematoma (site of fat harvest), which occurred in 23 patients (3.2%). Cerebrospinal fluid leak was present in 20 patients (2.8%), 15 of whom needed revision surgery. Other complications included VIth cranial nerve dysfunction in 12 cases (1.68%), subdural hematoma in 3 cases (0.4%), cerebellopontine angle hematoma in 4 cases (0.6%), cerebellar edema in 2 cases (0.28%), brainstem hematoma in 1 case (0.14%), transitory aphasia in 1 case (0.14%), and lower cranial nerve dysfunction in 1 case (0.14%). Mortality occurred in only one case (0.14%). Medical complications seldom occurred. The postoperative hospital stay ranged from 2 to 36 days, with an average of 6.4 days. The overall hospital stay diminished over time from 10.2 days in 1987 to 1990, to 4.9 days in 2001. There was a significant relation between hospital stay and tumor size, approach used, and presence/absence of complications. CONCLUSIONS: Perioperative complications in acoustic neuroma surgery do exist, but this study demonstrated how low the incidence is. The authors believe that the low percentage of complications is mainly attributable to the majority of operations being carried out in specialized clinics, where they are considered routine operations. They believe that following individualized approaches, depending on tumor size and on the preoperative function of the cranial nerves, is the proper way to reach a significant reduction in complications while maintaining a high percentage of total tumor removal. The results of this study, considered as a basis of comparison with other studies, will certainly be useful in preoperative patient counseling. PMID- 15129122 TI - The incidence of cerebrospinal fluid leak after vestibular schwannoma surgery. AB - OBJECTIVE: To review the incidence of cerebrospinal fluid leak after vestibular schwannoma removal reported in the literature. DATA SOURCES: MEDLINE and PubMed literature search using the terms "acoustic neuroma" or "vestibular schwannoma," and "cerebrospinal fluid leak" or "cerebrospinal fluid fistula" covering the period from 1985 to the present in the English language literature. A review of bibliographies of these studies was also performed. STUDY SELECTION: Criteria for inclusion in this meta-analysis consisted of the availability of extractable data from studies presenting a defined group of patients who had undergone primary vestibular schwannoma removal and for whom the presence and absence of cerebrospinal fluid leakage was reported. Studies reporting combined approaches were excluded. No duplications of patient populations were included. Twenty-five studies met the inclusion criteria. DATA EXTRACTION: Quality of the studies was determined by the design of each study and the ability to combine the data with the results of other studies. All of the studies were biased by their retrospective, nonrandomized nature. DATA SYNTHESIS: Significance (p < 0.05) was determined using the chi2 test. CONCLUSIONS: Cerebrospinal fluid leak occurred in 10.6% of 2,273 retrosigmoid surgeries, 9.5% of 3,118 translabyrinthine surgeries, and 10.6% of 573 middle fossa surgeries. The type of cerebrospinal fluid leak was not associated with surgical approach. Meningitis was significantly associated with cerebrospinal fluid leak (p < 0.05). Age and tumor size were not associated with cerebrospinal fluid leak. PMID- 15129123 TI - The evolution of surgical approaches for posterior fossa meningiomas. AB - OBJECTIVE: To evaluate surgical outcomes for the treatment of posterior fossa meningiomas at the House Ear Clinic from 1987 to 2001. We review our current treatment algorithm and present our postoperative outcomes with attention to facial and auditory neural preservation. STUDY DESIGN: Retrospective review. SETTING: Tertiary care center. PATIENTS: Medical records of 71 patients who underwent posterior fossa meningioma surgery at the House Ear Clinic were reviewed. INTERVENTION: All patients had surgical removal of their meningioma via translabyrinthine, transcochlear, retrosigmoid, extended middle fossa, or combined petrosal approaches by House Ear Clinic neurotologists and neurosurgeons. MAIN OUTCOME MEASURES: Preoperative and postoperative auditory and facial nerve function data were collected. Patient and tumor characteristics including presenting symptoms, completion of tumor resection, and complications secondary to surgery were also recorded. RESULTS: The most common presenting symptoms in this series were otologic, with hearing loss (61%), tinnitus (58%), and imbalance (58%) as the three most common. Gross total resection was achieved in 67 (94%) patients. Hearing-preservation surgery was attempted in 37 (52%) patients (68% via extended middle fossa or combined approach). Twenty-one patients with preoperative Class A hearing had follow-up audiometric data and 18 (86%) had serviceable hearing preserved. Excluding transcochlear craniotomies, 85% of patients had normal facial nerve function postoperatively. Cerebrospinal fluid leak (6%) was the most common complication. CONCLUSIONS: Advances in microsurgical techniques have greatly changed our management of patients with posterior fossa meningiomas. These changes have reduced postoperative morbidity. Specifically, use of the anterior and posterior petrosal approaches has facilitated facial and auditory neural preservation while not compromising the extent of tumor excision. PMID- 15129124 TI - Bilateral facial myokymia caused by fallopian canal dehiscence into the jugular bulb. PMID- 15129125 TI - Application of carbon dioxide and erbium:yttrium-aluminum-garnet lasers in inner ear surgery: an experimental study. AB - BACKGROUND: Surgery of the inner ear requires atraumatic techniques to preserve the sensory structures of the inner ear. With modern laser technology, surgery can be performed without mechanical contact, reducing the risk of direct mechanical trauma. However, energy transfer by laser light has the potential to induce damage by heating, pressure waves, or direct irradiation, depending on the properties of the laser and parameters of application. HYPOTHESIS: The application of laser systems in inner ear surgery may have an advantage over traditional techniques; the carbon dioxide laser in continuous mode with an automated scanning procedure and the erbium:yttrium-aluminum-garnet laser were compared with a mechanical technique, using a diamond drill. METHODS: A cochleostomy in the basal cochlear turn of guinea pigs was created. Thresholds in response to frequency-specific stimuli and clicks were established by recording compound action potentials, both before and after the procedure. RESULTS: The best results in terms of preservation of cochlear function were obtained with the diamond drill. However, a single ear had a complete loss after fracture of the cochlear wall. Mean threshold shifts observed with the carbon dioxide laser were slightly greater, showing mild high-frequency losses, although differences to the group of drilling were not statistically significant. Results with the erbium:yttrium-aluminum-garnet laser showed significantly higher degrees of hearing loss than the other two groups, predominantly in the high-frequency region. CONCLUSIONS: Mechanical opening of the inner ear using a microdrill can be performed with minimal hearing loss; however, it carries the risk of direct trauma to the inner ear. The carbon dioxide laser with a new scanning technology as a noncontact procedure is shown to be effective and safe. It can be regarded as a useful tool in inner ear surgery. The erbium:yttrium-aluminum-garnet laser has a greater potential to cause damage. PMID- 15129126 TI - Progression of facial nerve enhancement in idiopathic facial nerve paralysis. PMID- 15129127 TI - External auditory canal cholesteatoma. PMID- 15129132 TI - Bring hazy visitation policies into focus. PMID- 15129128 TI - Immunohistochemistry and histopathological assessment of the cholesteatoma of the ear. PMID- 15129135 TI - Sharp listening skills point staff in the right direction. AB - Evaluate how well you listen and communicate with staff members. PMID- 15129136 TI - Resource helps nurses help others. PMID- 15129137 TI - Medication management: saving lives one order at a time. AB - Get the latest update on the 2004 medication management's standards. PMID- 15129138 TI - Investigate thoroughly to avoid wrongful termination suits. AB - Review important investigative points to prevent wrongful termination of nurses. PMID- 15129139 TI - Nursing exploration attracts youth. PMID- 15129140 TI - Obesity: a rapidly expanding challenge. AB - : Explore the health implications of obesity, commonly performed surgical procedures, and postoperative care needs of bariatric surgery patients. PMID- 15129142 TI - Bouncing patients? Capacity issues and diversion status toy with efficient throughput. AB - : Explore several causes of decreased hospital capacity and review alternative patient flow strategies. PMID- 15129143 TI - Master the steps to performance improvement: plan, do, study, and act to enhance your facility's patient care initiatives. AB - When implementing process improvements, create awareness and include staff members most directly involved with the system. PMID- 15129146 TI - Smart pumps help crack the safety code. AB - Learn how manufacturers of infusion pumps and intravenous fluids technology aid hospitals in preventing intravenous medication errors. PMID- 15129151 TI - Strategies for replacing lost cochlear hair cells. AB - The auditory sensory epithelium is a mosaic composed of sensory (hair) cells and several types of non-sensory (supporting) cells. All these cells are highly differentiated in their structure and function. Mosaic epithelia (and other complex tissues) are generally formed by differentiation of distinct and specialized cell types from common progenitors. Most types of epithelial tissues maintain a population of undifferentiated (basal) cells which facilitate turnover (renewal) and repair, but this is not the case for the organ of Corti in the cochlea. Therefore, when cochlear hair cells are lost they cannot be replaced. Consequently, sensorineural hearing loss is permanent. In designing therapy for sensorineural deafness, the most important task is to find a way to generate new cochlear hair cells to replace lost cells. PMID- 15129152 TI - Optical imaging of binaural interaction in multiple fields of the guinea pig auditory cortex. AB - Locating the source of a sound is an important function of the auditory system and interaural intensity differences are one of the most important cues. To study the functional pathways of sound localisation processing in the auditory cortex, activity in multiple fields of the guinea pig auditory cortex during stimulation with interaural intensity differences was studied using optical imaging with a voltage-sensitive dye. Of the auditory core (primary and dorsocaudal) and the belt fields which surround them, the posterior and ventroposterior belt fields were the most sensitive to interaural intensity differences. This suggests that the caudal pathway of the auditory cortex is involved in sound localisation. PMID- 15129153 TI - Spatiotemporal brain activation pattern during word/picture perception by native Koreans. AB - The purpose of this study is to explore spatiotemporal brain activation patterns during perception of words from three different languages (Korean, English, Chinese) and pictures. Using 64 channel event-related potential (ERP) recording and source localization using distributed source model, we investigated, with high temporal resolution, whether similar or different spatiotemporal patterns of brain activation are involved in the perception of words of different languages and/or pictures. Experimental results seem to corroborate left hemispheric dominance in language processing, and temporal/spatial characteristics in word perception revealed by previous ERP and neuroimaging studies. Observed differences in spatial pattern of activation at specific time periods between English and Korean, and Korean and Chinese, could be explained in terms of required visual pattern analysis due to the orthographic characteristics of each language. PMID- 15129154 TI - Functional recovery in chronic paraplegia after bone marrow stromal cells transplantation. AB - Previous reports showed the therapeutic effect of transplants of bone marrow stromal cells (BMSC) after incomplete traumatic spinal cord lesions. We studied the effect of this form of therapy in chronically paraplegic Wistar rats due to severe spinal cord injury (SCI). Rats were subjected to weight-drop impact causing paraplegia, and BMSC or phosphate buffered saline (PBS) was injected into spinal cord 3 months after injury. Functional outcome was measured using the Basso-Beattie-Bresnehan score until sacrifice of the animals, 4 weeks after transplantation. At this time, samples of spinal cord tissue were studied histologically. The results showed a clear and progressive functional recovery of the animals treated with BMSC transplantation, compared to controls. Grafted BMSC survived into spinal cord tissue, forming cell bridges within the traumatic centromedullary cavity. In this tissue, cells expressing neuronal and astroglial markers can be seen, together with a marked ependymal proliferation, showing nestin-positivity. These findings suggest the utility of BMSC transplantation in chronically established paraplegia. PMID- 15129155 TI - Affective picture perception: gender differences in visual cortex? AB - Activity in extrastriate visual cortex is greater when people view emotional relative to neutral pictures. Prior brain imaging and psychophysiological work has further suggested a bias for men to react more strongly to pleasant pictures, and for women to react more strongly to unpleasant pictures. Here we investigated visual cortical activity using fMRI in 28 men and women during picture viewing. Men and women showed reliably greater visual cortical reactivity during both pleasant and unpleasant pictures, relative to neutral, consistent with the view that the motivational relevance of visual stimuli directs attention and enhances elaborative perceptual processing. However, men did show greater extrastriate activity than women specifically during erotic picture perception, possibly reflecting a gender-specific visual mechanism for sexual selection. PMID- 15129156 TI - In situ detection of apolipoprotein E epsilon 4 in archival human brain. AB - A monoclonal antibody specific to apolipoprotein E 4 (apoE4) was applied immunohistochemically to archival human brain tissue. The examined 30 cases comprised four epsilon/epsilon4, 10 epsilon3/epsilon4, one epsilon2/epsilon4, 10 epsilon3/epsilon3 and five epsilon2/epsilon3 genotypes. The anti apoE4 antibody visualized senile plaques, neurofibrillary tangles and reactive astrocytes, as well as serum in the blood vessels and vascular smooth muscle cells in the cases of epsilon4. Moreover, the staining intensity was stronger in the cases carrying the epsilon4 homozygosity than in those cases of epsilon4 heterozygosity. Specific immunoreactivity was not obtained in those cases not carrying the epsilon4 allele. This method will allow in situ detection of apoE epsilon4 and contribute to studies of the effect of epsilon4 on Alzheimer's disease. PMID- 15129157 TI - Radionuclide imaging of mesenchymal stem cells transplanted into spinal cord. AB - In vivo tracking of stem cells implanted to spinal cord by radionuclide imaging was investigated. The high expression of transferrin receptor on human mesenchymal stem cells (hMSCs) was verified by flow cytometry, radioligand binding and immunofluorescence. Radiolabelled transferrin was chosen as a tracer for scintigraphic imaging of the hMSCs transplanted into spinal cord of rabbits. Comparative experiments with radiolabelled human serum albumin as tracer and PBS as graft as well as ex vivo autoradiography demonstrated the specific uptake of radiolabelled transferrin of hMSCs. hMSCs could be detected in vivo with radiolabelled transferrin targeting at cellular transferrin receptors at an early stage after transplantation into spinal cord. PMID- 15129158 TI - Delayed neurotrophin treatment supports auditory neuron survival in deaf guinea pigs. AB - As key factors in the development and maintenance of the auditory system, neurotrophins can prevent auditory neuron degeneration when applied within three to five days of deafening. We tested each of the neurotrophins BDNF, NT-3, NT-4/5 and NGF for their ability to support auditory neuron survival following a two week period of deafness in guinea pigs, when approximately 15% auditory neuron degeneration has already occurred. Although delayed, the treatment with each neurotrophin prevented further degeneration with similar efficacy. PMID- 15129159 TI - FOS expression in orexin neurons following muscimol perfusion of preoptic area. AB - Unilateral microdialysis-perfusion of the preoptic area with 50 microM muscimol decreased the sleep period of rats to less than 3% of the baseline value over a 90 min period before death (p = 0.018 by Wilcoxon signed-rank test). These rats showed the expression of FOS in 36% of the orexin neurons located in the perifornical/lateral hypothalamic areas on the side ipsilateral to the perfusion site, but in only 3% of the orexin neurons on the side contralateral to it (p = 0.018 by Wilcoxon signed-rank test). These results suggest that subpopulations of the preoptic neurons give an inhibitory tone to the activities of the orexin neurons in the perifornical/lateral hypothalamic areas. PMID- 15129160 TI - 1/f-type fluctuation in human visuomotor transformation. AB - In the absence of vision of the limb, movements toward a visual target exhibit substantial errors which are considered to originate mainly in the visuomotor transformation process. To determine the time-dependent property of human visuomotor transformation, we investigated the error sequences in movements toward visual target using scaling analyses. When subjects could see their controlling limb, the error sequences could not be distinguished from a random sequence. On the other hand, when the controlling limb was invisible, the error sequences were not random in order, but exhibited 1/f-type time correlation. This finding that the variation in human visuomotor transformation shows 1/f-type fluctuation provides a significant index for mathematical modeling and system identification in human visuomotor control. PMID- 15129161 TI - Deep brain stimulation creates an informational lesion of the stimulated nucleus. AB - Deep brain stimulation (DBS) is an effective treatment for movement disorders, but the mechanisms are unclear. DBS generates inhibition of neurons surrounding the electrode while simultaneously activating the output axons of local neurons. This dual effect does not explain two hallmarks of DBS effectiveness: symptom relief is dependent on using a sufficiently high-stimulation frequency, and clinical effects are analogous to those produced by lesion. The effect of DBS at different frequencies on the output of intrinsically active neurons was studied using computational models. DBS produced frequency-dependent modulation of the variability of neuronal output, and above a critical frequency stimulation resulted in regular output with zero variance. The resulting loss of information offers an explanation for the two hallmarks of DBS effectiveness. PMID- 15129162 TI - GABA content in the retina of pigmented and albino rats. AB - Reduction of the melanin precursor DOPA associated with albinism leads to spatiotemporal disturbances in retinal neurogenesis and thus seems to be responsible for numerous neuronal alterations found in albino retinae. To investigate whether these cellular alterations are reflected in retinal neurotransmitter concentrations we compared the levels of GABA and glutamate in the retina of adult pigmented Long Evans and albino Wistar rats using reversed phase-liquid chromatography (RP-HPLC). When normalized to retinal weight, GABA levels showed a statistically insignificant trend to be lower and glutamate values to be higher in albinos than in pigmented animals. The ratio of glutamate to GABA was significantly higher in albino than in pigmented retinae. As numerous studies have shown that the balance between GABA and glutamate plays a crucial role for establishing direction selectivity, these results are discussed in relation to direction selectivity and defects in the optokinetic system of albinos. PMID- 15129163 TI - Triazolam suppresses the induction of hippocampal long-term potentiation. AB - Benzodiazepines are sedative hypnotics that produce marked anterograde amnesia in humans. These pharmacological properties are thought to result from the potentiation of GABA-A receptor function and subsequent attenuation of long-term potentiation (LTP), however many reports have suggested this is not the case for triazolam. Using electrophysiological recordings in a cell line expressing recombinant GABA-A receptors, we confirm that triazolam is an efficacious positive allosteric modulator of GABA-A receptors. Triazolam also slowed the decay of spontaneous inhibitory synaptic currents, reduced the amplitude of fEPSPs elicited during a theta burst and reduced the magnitude of LTP in hippocampal CA1 neurones in vitro. These data show that triazolam modifies LTP induction consistent with an enhancement of GABA-A receptor function via activation of the allosteric benzodiazepine-site. PMID- 15129164 TI - MK-801 does not prevent development of ischemic tolerance in rat brain. AB - Tolerance against ischemia can be induced in the CA1 region of the hippocampus of the brain. In gerbils tolerance evolvement is blocked by the NMDA-antagonist MK 801. To examine this mechanism in rats, MK-801 was administered i.p. 1 h prior to tolerance inducing ischemia. Body temperature and activity were monitored before and after ischemia, and show that MK-801 results in hyperthermia immediately after the injection, the post-ischemic body temperature remain elevated until 5 h post-ischemia in spite of the animals being less active than control animals. Histology shows that pre-treatment with MK-801 does not affect the CA1 neuronal density, and we thus conclude that for the used rat model, MK-801 does not affect development of ischemic tolerance. PMID- 15129165 TI - Interleukin-10 expression in lipopolysaccharide-activated microglia is mediated by extracellular ATP in an autocrine fashion. AB - Immune cells have been shown to release ATP into the extracellular space to provide auto- and paracrine purinergic modulation of immune and inflammatory responses. In the present study, we demonstrate that ATP released from lipopolysaccharide (LPS)-stimulated microglia induces interleukin-10 (IL-10) expression in an autocrine manner. The expression as well as secretion of IL-10 by LPS-stimulated microglia was completely inhibited by apyrase, ATP-hydrolyzing enzyme, whereas tumor necrosis factor-alpha (TNF-alpha) expression was unaffected. LPS-activated microglia rapidly released a low concentration of ATP (10-20 nM) into the medium, and the nanomolar range of extracellular ATP, ADP, adenosine 5'-O-(3-thiotriphosphate) (ATP-gamma-S), and adenosine 5'-O-(2 thiodiphosphate) (ADP-beta-S) induced IL-10 secretion from microglia in a dose dependent manner. These results suggest that ATP released from LPS-activated microglia and/or a metabolite of ATP (ADP) may induce IL-10 expression through P2Y purinergic receptors. PMID- 15129166 TI - Local acute application of BDNF in the lesioned spinal cord anti-inflammatory and anti-oxidant effects. AB - We studied the early anti-inflammatory and anti-oxidant effects of local application of BDNF after dorsal spinal cord transection in the adult rat. Both the distribution and accumulation of neutrophils and microglial cells in and around the lesion site (inflammatory response) and the accumulation of lipid peroxidation product 4-hydroxynonenal (HNE; oxidative damage) around the lesion was examined using immunohistochemical techniques. We demonstrate that BDNF application affects the microglial response in and around the lesion and results in a reduced lipid peroxidation as shown by HNE-immunoreactive staining around the lesion 48 h post-injury. The early anti-inflammatory and anti-oxidant effects of local BDNF-application into the lesioned spinal cord may contribute to the observed decreased loss of locomotor function of the hindlimbs 2 days after injury. PMID- 15129167 TI - SH-SY5Y neuroblastoma cells: a model system for studying biosynthesis of NAAG. AB - N-Acetylaspartylglutamate (NAAG) is a neuropeptide that is thought to modulate neurotransmitter release through pre-synaptic mGluR3 receptors. Despite years of research into NAAG biochemistry, almost nothing is known about NAAG biosynthesis. To date, NAAG biosynthesis has only been demonstrated conclusively in explanted animal neural tissues, including frog retina, rat dorsal root ganglia and crayfish nerve cord, but not in human cells or tissues. We show here that a human neuroblastoma cell line, SH-SY5Y, provides a good model system for the study of NAAG biosynthesis. Radiolabled NAAG synthesis occurred using both L-[3H]glutamic acid and L-[3H]glutamine as precursors, with glutamine being the preferred substrate. Differentiation of SH-SY5Y cells with retinoic acid resulted in decreased radiolabel incorporation into NAAG, whereas differentiation with nerve growth factor did not affect radiolabel incorporation. PMID- 15129168 TI - Adenoviral expression of CREB protects neurons from apoptotic and excitotoxic stress. AB - In this study we have used a molecular approach to manipulate CREB gene expression to study its role in the regulation of neuronal cell death. To achieve this, adenoviral (Ad) vectors encoding EGFP, CREB, and a powerful CREB dominant negative, known as A-CREB were constructed. The over-expression of CREB but not A CREB was found to protect primary hippocampal neurons from staurosporine-induced apoptosis, glutamate induced excitotoxicity and exposure to an in vitro ischaemic stress. Hence, manipulating CREB-regulated pathways may provide a means of delaying or preventing the neuronal cell death associated with ischaemic related injury, and in neurodegenerative diseases such as Huntington's and Alzheimer's disease. PMID- 15129169 TI - Role of Ca2+ in induction of neurotransmitter-related gene expression by butyrate. AB - We examined the effect of butyrate on neurotransmitter-related gene expression and calcium homeostasis in PC12 cells. Pretreatment with Ca2+ chelators (EGTA or BAPTA-AM) attenuated the butyrate-triggered accumulation of TH and ppEnk mRNA indicating that Ca2+ plays a role in butyrate-induced regulation of neuronal genes. Butyrate alone did not alter intracellular Ca2+ levels as determined by Fura-PE3 fluorescence; however, pretreatment with butyrate (18-24 h) reduced the first Ca2+ peak and prevented the second sustained rise in [Ca2+]i as induced by nicotine or ryanodine. In contrast, butyrate had no effect on Ca2+ transients when added shortly before or during nicotine or ryanodine stimulation. These results suggest that chronic butyrate exposure can modulate cell responses by affecting intracellular Ca2+ signaling. PMID- 15129170 TI - Multidrug resistance protein 1-mediated transport of saquinavir by microglia. AB - Regulation of CNS distribution of the human immunodeficiency virus (HIV) protease inhibitor saquinavir may involve ATP-dependent membrane-bound efflux transport proteins that are expressed in several brain cellular compartments. We recently characterized molecular and functional expression of one such transporter, multidrug resistance protein-1 (MRP1) in microglia, the primary brain cellular target of HIV. In the present study, we further examine subcellular localization of MRP1 in a microglia cell line (MLS-9) using immunogold cytochemistry and directly demonstrate MRP1-mediated export of saquinavir. MRP1 localized primarily to the plasma membrane of the MLS-9 cells. [14C]Saquinavir efflux by MLS-9 monolayers was inhibited by well-established MRP1 inhibitors. These results indicate that MRP1 contributes, in part, to the overall low permeation of protease inhibitors in the brain. PMID- 15129171 TI - Effect of halothane on the release of [Ca2+]i in dorsal root ganglion neurons. AB - We investigated the effect of the volatile anaesthetic halothane on [Ca2+]i of dorsal root ganglion neurons. Halothane was able to increase [Ca2+]i in those neurons in a dose-dependent manner and independent of extracellular calcium. However, halothane action was inhibited by BAPTA-AM, suggesting the involvement of intracellular calcium stores. Dantrolene, an inhibitor of ryanodine-sensitive calcium stores had no effect while 2-APB, an inhibitor of IP3-sensitive calcium store reduced by 78% the halothane-evoked increase on [Ca2+]i. These data suggests that halothane increased [Ca2+]i of ganglion neurons through calcium release from IP3-sensitive calcium store. One possible consequence of the halothane action is to alter presynaptic activity and signaling pathways that influence neurotransmission. PMID- 15129172 TI - Effects of GSM electromagnetic field on the MEG during an encoding-retrieval task. AB - Potential effects of GSM 1800 electromagnetic fields (EMF) on verbal memory encoding were investigated by recording event-related magnetic fields (ERMF) from the brain during subsequent memory retrieval. Twelve normal subjects participated in the study. After encoding words from a study list presented in the first phase they had to discriminate old from new words mixed together in a test list presented during the second phase. All subjects performed two experimental sessions, one with exposure to EMF during the study phase, and one without. Exposure to EMF changed an early (350-400 ms) task-specific component of the ERMF indicating an interference of EMF and item encoding. Behavioural measures were not significantly affected. Adverse health effects cannot be derived from these data. PMID- 15129173 TI - Cerebral hemodynamics and processing speed during category learning. AB - The Wisconsin Card Sorting Test (WCST) is a commonly used paradigm of category learning with two alternating and distinct processes, maintaining set and set shifting that have been characterized by functional transcranial Doppler sonography (fTCD) of the basal cerebral arteries. Further, repeated WCST administrations resulted in improved speed of solution. This study addressed the question of whether optimised speed during maintaining set or set shifting is associated with changes of cerebral hemodynamics as measured by bilateral fTCD. During maintaining set, improved speed was associated with increased peak mean cerebral blood flow velocity (CBFV) of the anterior cerebral arteries. These results suggest a common functional substrate between optimised speed of solution and cerebral hemodynamics during maintaining set. PMID- 15129174 TI - Prismatic adaptation reduces biased temporal order judgements in spatial neglect. AB - Visuomotor adaptation to rightward-deviating optical-wedge prisms reduces the clinical manifestations of spatial neglect after right hemisphere damage. We investigated whether this beneficial effect of prism adaptation is due to attenuation of the ipsilesional attentional bias that is common in spatial neglect. Five right hemisphere patients performed visual temporal order judgements before and after visuomotor adaptation to 15 degrees rightward deviating prisms. The magnitude of patients' ipsilesional attentional bias on the temporal order judgement task was significantly reduced following adaptation. By contrast, the temporal order judgements of normal participants did not change following adaptation to either leftward- or rightward-deviating prisms. The findings suggest that prism adaptation helps to rebalance the distribution of spatial attention following right hemisphere damage. PMID- 15129175 TI - Glides in speech fundamental frequency are reflected in the auditory N1m response. AB - The cortical dynamics underlying the perception of constant and gliding speech fundamental frequency (F0) was investigated in 10 subjects using magnetoencephalography (MEG). The stimuli comprised vowels having either constant, ascending or descending F0s and tones of corresponding frequencies, matched with the vowels in intensity or loudness. The amplitude of the N1m response was highly sensitive to F0 variation embedded in vowels and insensitive to corresponding variation in tones. The latency of the N1m elicited by the tones with respect to vowels was significantly delayed. Thus, the speech-specific behavior of the N1m arises out of cortical sensitivity to the acoustic structure of voiced speech, that is to the F0 and its harmonics, which underlie the perception of pitch and intonation in speech. PMID- 15129176 TI - Lexical memory search during N400: cortical couplings in auditory comprehension. AB - In this study, we applied partial-directed EEG-coherence analysis to assess regional changes in neuronal couplings and information transfer related to semantic processing. We tested the hypothesis whether (and which) processing differences between spoken words and pseudowords are reflected by changes in cortical networks within the time window of a specific event related potential (ERP) component, the N400. Fourteen native speaking German subjects performed a lexical decision paradigm, while being confronted sequentially with two-syllabic nouns and phonologically legal pseudowords. Using ERP analysis, we defined the time window of the N400 effect, known to reflect semantic processing, and, subsequently, we examined the coupling differences. Lexico-semantic memory search appears to be subserved by a network between temporal, parietal and frontal areas, particularly restricted to the left hemisphere. PMID- 15129177 TI - Association of EGF polymorphism with schizophrenia in Finnish men. AB - Some recent data suggest that epidermal growth factor (EGF) protein levels are altered in the brain of schizophrenic patients. In addition, a novel polymorphism of the EGF gene is associated with enhanced production of EGF in vitro. We conducted a retrospective study to explore the impact of EGF polymorphism on factors associated with schizophrenia. The sample consisted of 94 patients with schizophrenia who had either responded to treatment with conventional neuroleptics or who were considered non-responders. The control sample consisted of 98 blood donors. In our sample, the G allele was associated with schizophrenia in male patients (OR = 3.594 (95% CI 1.347-9.591), p = 0.008). The G allele was also associated with a later age at onset in male patients with schizophrenia. However, no association was found between treatment response and EGF polymorphism. PMID- 15129178 TI - Expression of tetrodotoxin-sensitive and resistant sodium channels by rat melanotrophs. AB - Rat melanotrophs fire Na+ and Ca2(+)-dependent action potentials. Whereas the molecular identity of Ca2+ channels expressed by these cells is well documented, less is known about Na channels. We characterize the expression of seven sodium channel alpha-subunit and the beta1- and beta2-subunit mRNAs. The tetrodotoxin resistant Nav1.8 and Nav1.9 alpha subunit mRNAs are detected in the newborn intermediate lobe and in cultured melanotrophs. Electrophysiological recordings further demonstrate the expression of both tetrodotoxin-sensitive and tetrodotoxin-resistant currents by dissociated melanotrophs. Moreover, activated sodium channels are able to elicit intracellular calcium waves, both in the absence or in the presence of tetrodotoxin. This work shows that rat melanotrophs express functional tetrodotoxin-resistant sodium channels, whose activation can lead to the generation of intracellular calcium waves. PMID- 15129179 TI - Anoxia tolerant brains. AB - While medical science has struggled to find ways to counteract anoxic brain damage with limited success, evolution has repeatedly solved this problem. The best-studied examples of anoxia-tolerant vertebrates are the crucian carp and some North American Freshwater turtles. These can survive anoxia for days to months, depending of temperature. Both animals successfully fight any major fall in brain ATP levels, but the strategies they use to accomplish this are quite divergent. The anoxic turtle suppresses brain activity to such a degree that it becomes virtually comatose. The underlying mechanisms involve closing down ion conductances and releasing GABA and adenosine. By contrast, the crucian carp remains active in anoxia, although it suppresses selected brain functions, and avoids lactate self-poisoning by producing an exotic anaerobic end-product. These animals provide unique models for studying anoxic survival mechanisms both on a molecular and physiological level. PMID- 15129180 TI - Intracerebral hemorrhage in mice: model characterization and application for genetically modified mice. AB - Gene knockout or transgenic animals may assist in elucidating the mechanisms of brain injury after intracerebral hemorrhage (ICH). However, almost all commercially available transgenic or knockout animals are mice. The purpose of this study was to develop an ICH model in mice and to investigate the influence of gender and complement C5 genetic differences on outcome after ICH. Male and female C57BL/6 mice and C5-deficient and -sufficient control mice were anesthetized and then received an injection of 30 microL autologous whole blood into the right basal ganglia. Brain water content was studied at 1 and 3 days after ICH. Behavioral tests (fore-limb use asymmetry and corner turn test) were performed at 1, 3, 7, 14, 21, or 28 days after ICH. In male mice, brain water content was significantly increased in the ipsilateral basal ganglia 1 and 3 days after ICH, compared with saline injection controls (P < 0.01). There were marked neurological deficits 1 and 3 days after ICH, with progressive recovery over 28 days. In contrast, although brain edema and behavioral deficits were similar at 1 day after ICH in female and male mice, female mice showed reduced edema at 3 days and a faster recovery of behavioral deficits after ICH. 17 beta-estradiol treatment in male mice markedly reduced ICH-induced edema (P < 0.01). Brain water content was significantly increased in C5-deficient mice compared with C5 sufficient at 3 days after ICH (P < 0.05). These findings suggest that the mouse ICH model is a reproducible and feasible model. These results also suggest that gender and complement C5 are factors affecting brain injury after ICH. PMID- 15129181 TI - Outcome of acutely ischemic brain tissue in prolonged middle cerebral artery occlusion: a serial positron emission tomography investigation in the baboon. AB - Thrombolysis within 3 to 6 hours of symptom onset is recommended therapy for acute middle cerebral artery (MCA) stroke, but recent imaging studies in humans suggest that the penumbra may last much longer in some patients. It is therefore important to study the events that take place with occlusions that last longer than 6 hours. Based upon positron emission tomography (PET), the tissue with high oxygen extraction fraction (OEF) is at risk of infarction. In a previous sequential PET study in anesthetized baboons, we documented that when reperfusion was initiated at 6 hours after MCA occlusion, the region with the acutely highest OEF was not incorporated within the final magnetic resonance imaging (MRI) defined infarct, suggesting reperfusion prevented such demise. In agreement with this hypothesis, we report here using the same sequential PET paradigm with final chronic-stage volume MRI that a 20-hour MCA occlusion resulted in, on average, 36% of the highest OEF area being recruited into the final infarct. We also found that the portion of the highest OEF area that went on to infarct had at the earliest time-point significantly lower cerebral blood flow and cerebral oxygen metabolism (mean reductions relative to unoccluded side, 56% and 32%, respectively) than the portion that did not (41% and 11%, respectively) and that some reperfusion occurred in the latter at second time-point, that is, before recanalization. Thus, apart from duration of occlusion, the fate of the at-risk tissue is predicated by the initial severity of the ischemia as well as by early secondary events such as partial spontaneous reperfusion. PMID- 15129182 TI - Dependency of cortical functional hyperemia to forepaw stimulation on epoxygenase and nitric oxide synthase activities in rats. AB - Individual inhibition of nitric oxide (NO) synthase and cytochrome P450 (CYP) epoxygenase activity attenuates cortical functional hyperemia evoked by whisker stimulation. The objectives of the present study were to determine (1) if administration of epoxygenase inhibitors attenuates cortical functional hyperemia by using a different modality of sensory activation (i.e., electrical stimulation of the rat forepaw), (2) if epoxygenase inhibition has an additive effect with NO synthase inhibition on the flow response, and (3) the cellular localization of the epoxygenase CYP2C11 in cerebral cortex. In six groups of anesthetized rats, the cortical surface was superfused for 90 minutes with (1) vehicle; (2) 1-mmol/L Nomega-nitro-L-arginine (L-NNA), to inhibit NO synthase activity; (3) 20 micromol/L N-methylsulfonyl-6-(2-propargyloxyphenyl)hexanamide (MS-PPOH), a substrate inhibitor of P450 epoxygenase; (4) MS-PPOH plus L-NNA; (5) 20 micromol/L miconazole, a reversible inhibitor at the heme site of P450 epoxygenase; and (6) miconazole plus L-NNA. The percent increases in laser Doppler perfusion over primary sensory cortex during 20-second forepaw stimulation were reduced by 44% to 64% in all drug-treated groups. The addition of L-NNA to MS-PPOH produced no additional reduction (64%) compared with MS-PPOH alone (64%) or L-NNA alone (60%). The addition of L-NNA to miconazole also produced no additional reduction in the flow response. In situ hybridization of CYP2C11 mRNA showed localization in astrocytes, including those adjacent to blood vessels. Thus, activity of both epoxygenase, presumably localized in astrocytes, and NO synthase is required for generating a complete cortical hyperemic response evoked by electrical forepaw stimulation. The lack of additional blood flow attenuation with the combination of the NO synthase and the distinct epoxygenase inhibitors suggests that the signaling pathways do not act in a simple parallel fashion and that other mediators may be involved in coupling cortical blood flow to neuronal activation. PMID- 15129183 TI - Spatiotemporal quantification of cerebral blood flow during functional activation in rat somatosensory cortex using laser-speckle flowmetry. AB - Laser-speckle flowmetry was used to characterize activation flow coupling after electrical somatosensory stimulation of forepaw and hindpaw in the rat. Quantification of functional activation was made with high transverse spatial (microm) and temporal (msec) resolution. Different activation levels and duration of stimulation were quantitatively investigated, and were in good agreement with previous laser-Doppler measurements. Interestingly, the magnitude but not the overall shape of the response was found to scale with stimulus amplitude and the distance from the activation centroid. The results provide new insights about the spatial characteristics of cerebral blood flow response to functional activation, and the method should lead to improved understanding of the coupling of neuronal activity and hemodynamics under normal and pathologic conditions. PMID- 15129184 TI - Impaired vascular reactivity of isolated rat middle cerebral artery after cortical spreading depression in vivo. AB - Cortical spreading depression (CSD) is accompanied by hyperemia followed by long lasting hypoperfusion and impaired cerebrovascular reactivity. The authors show that vasodilation to extraluminal acidosis (pH 7.0) and increased concentrations of extraluminal potassium (12, 20, 40 mmol/L) was significantly reduced in isolated rat middle cerebral arteries after CSD in vivo before the artery was isolated, compared with sham-operated controls. Application of 80-mmol/L potassium induced vasoconstriction after CSD. Therefore, the impairment of vascular reactivity after CSD in vivo occurs, at least in part, at the vascular level itself. PMID- 15129185 TI - In vivo binding behavior of dopamine receptor agonist (+)-PD 128907 and implications for the "ceiling effect" in endogenous competition studies with [(11)C]raclopride-a positron emission tomography study in Macaca mulatta. AB - In in vivo positron emission tomography (PET) studies, dopamine that is released secondary to amphetamine administration appears unable to achieve a receptor occupancy that is significantly higher than 50% ("ceiling effect"). Also with exogenous agonists no studies have reported a higher than 50% occupancy. To investigate the feasibility of exceeding 50% occupancy in vivo with a dopamine receptor agonist we administered D2/D3 agonist (+)-PD 128907 over an extensive dose range. Two anesthetised Macaca mulatta males were used in a bolus-infusion protocol for [(11)C]raclopride. (+)-PD 128907 was administered as an intravenous challenge during separate PET scans in a dose range of 10 to 10000 nmol/kg. Occupancy by (+)-PD 128907 was estimated by comparing the binding before and after challenge. In a striatal region of interest, receptor occupancy by (+)-PD 128907 increased in an orderly dose-dependent manner to a maximum of at least 85%. This is the first indication that virtually all dopamine D2/D3 receptors in the striatum are in principle accessible to agonist binding. In the case of dopamine a number of protective mechanisms may be responsible for the ceiling effect. PMID- 15129186 TI - Sustained blockade of brain AT1 receptors before and after focal cerebral ischemia alleviates neurologic deficits and reduces neuronal injury, apoptosis, and inflammatory responses in the rat. AB - In the present study, we investigate whether a long-term blockade of brain AT1 receptors in male Wistar rats before and after ischemic injury exerts neuroprotective effects and modulates apoptosis and inflammatory responses, which are associated with the post-ischemic progression of brain damage. The AT1 receptor antagonist irbesartan was continuously infused intracerebroventricularly using osmotic minipumps over a 5-day period before and for 3 or 7 days after middle cerebral artery occlusion (MCAO) for 90 minutes. Neurologic status was evaluated daily, starting 24 hours after MCAO. After MCAO (3 and 7 days), brains were removed for the measurement of infarct size and immunohistochemical evaluation of apoptosis and accumulation of reactive microglia and macrophages. Treatment with irbesartan before ischemia improved motor functions, whereas post ischemic treatment improved sensory functions. Blockade of brain AT1 receptors reduced the infarct size on days 3 and 7 after MCAO. In the peri-infarct cortex, irbesartan treatment decreased the number of apoptotic cells on day 3 and attenuated the invasion of activated microg-lia and macrophages on days 3 and 7 after ischemia. Long-term blockade of brain AT1 receptors improves the recovery from cerebral ischemia. Antiapoptotic mechanisms and inhibition of post-ischemic inflammation are involved in the AT1 receptor blockade-induced neuroprotective effects in ischemic brain tissue. PMID- 15129187 TI - Characterization of BrdU-positive neurons induced by transient global ischemia in adult hippocampus. AB - Neurogenesis in the brain continues throughout life and is promoted by brain insults including ischemia. There is no critical conclusion, however, about whether proliferated cells acquire neuronal function after ischemia. Transient global ischemia was produced by a four-vessel occlusion procedure in rats (n = 54). To label proliferative cells, rats were administrated with a single dose of 5-bromo-2'-deoxyuridine (BrdU) at 4, 6, 8, 10, 13, or 15 days after ischemia. Increases in BrdU-positive cells were detected in the hippocampal dentate gyrus at 5, 7, and 9 days after ischemia. To determine the phenotype of BrdU-positive cells, BrdU was administrated twice daily for 3 consecutive days during 6 to 8 days after ischemia. A basic helix-loop-helix transcription factor NeuroD at 7 and 14 days and an immature migrating neuronal marker doublecortin at 14 days after ischemia were expressed transiently in proliferative cells. These proliferative cells after ischemia differentiated to the phenotype of neuron at 28 days after ischemia. Furthermore, BrdU-positive neurons showed phosphorylation of extracellular signal-regulated kinase (ERK) by intracerebroventricular injection of N-methyl-D-aspartate (NMDA) at 28 and 56 days after ischemia as seen in surrounding mature neurons. The number of BrdU-positive neurons, which responded to NMDA stimulation, increased with time after ischemia and was greater than that of sham-operated animals. The present study provides evidence for in vivo ERK phosphorylation in response to NMDA stimulation of BrdU-positive neurons in the adult hippocampus after transient forebrain ischemia. PMID- 15129188 TI - Pharmacological manipulations of ATP-dependent potassium channels and adenosine A1 receptors do not impact hippocampal ischemic preconditioning in vivo: evidence in a highly quantitative gerbil model. AB - Ischemic preconditioning models have been characterized in brain, heart, and other tissues, and previous pharmacologic studies have suggested an involvement of adenosine and ATP dependent potassium (KATP) channels in such tolerance phenomena. This question was reexamined in a reproducible gerbil model in which the duration of ischemic depolarization defined the severity of preconditioning and test insults. Agents studied were glibenclamide, a blocker of KATP channels; 1,3-dipropyl-8-cyclopentylxanthine (DPCPX), an adenosine A1 receptor antagonist; and N6-cyclopentyladenosine (CPA), an A1 agonist. Intraventricular glibenclamide injections aggravated neuron damage after brief priming insults, in parallel with a dose-dependent prolongation of ischemic depolarization. However, the depolarization thresholds for ischemic neuronal injury were identical in vehicle- and glibenclamide-treated animals, and glibenclamide did not affect preconditioning when equivalent insult severity was maintained during priming insults. Neither DPCPX nor CPA had any effect on the onset or duration of depolarization after intraperitoneal injection in this model, and neither drug affected neuron damage. In the case of CPA, it was necessary to maintain temperature for 4 to 6 hours of recirculation to avoid significant confounding hypothermia. These results fail to support a direct involvement of A1 receptors or KATP channels during early stages in the development of ischemic tolerance in vivo, and emphasize the need for robust, well-controlled, and quantitative models in such studies. PMID- 15129189 TI - The "dark side" of endocannabinoids: a neurotoxic role for anandamide. AB - Endocannabinoids, including 2-arachidonoylglycerol and anandamide (N arachidonoylethanolamine; AEA), have neuroprotective effects in the brain through actions at CB1 receptors. However, AEA also binds to vanilloid (VR1) receptors and induces cell death in several cell lines. Here we show that anandamide causes neuronal cell death in vitro and exacerbates cell loss caused by stretch-induced axonal injury or trophic withdrawal in rat primary neuronal cultures. Administered intracerebroventricularly, AEA causes sustained cerebral edema, as reflected by diffusion-weighted magnetic resonance imaging, regional cell loss, and impairment in long-term cognitive function. These effects are mediated, in part, through VR1 as well as through calpain-dependent mechanisms, but not through CB1 receptors or caspases. Central administration of AEA also significantly upregulates genes involved in pro-inflammatory/microglial-related responses. Thus, anandamide produces neurotoxic effects both in vitro and in vivo through multiple mechanisms independent of the CB1 receptor. PMID- 15129190 TI - Changes in cerebral blood flow during cerebrospinal fluid pressure manipulation in patients with normal pressure hydrocephalus: a methodological study. AB - The combination of cerebral blood flow measurement using (15)O-water positron emission tomography with magnetic resonance coregistration and CSF infusion studies was used to study the global and regional changes in CBF with changes in CSF pressure in 15 patients with normal pressure hydrocephalus. With increases in CSF pressure, there was a variable increase in arterial blood pressure between individuals and global CBF was reduced, including in the cerebellum. Regionally, mean CBF decreased in the thalamus and basal ganglia, as well as in white matter regions. These reductions in CBF were significantly correlated with changes in the CSF pressure and with proximity to the ventricles. A three-dimensional finite element analysis was used to analyze the effects on ventricular size and the distribution of stress during infusion. To study regional cerebral autoregulation in patients with possible normal pressure hydrocephalus, a sensitive CBF technique is required that provides absolute, not relative normalized, values for regional CBF and an adequate change in cerebral perfusion pressure must be provoked. PMID- 15129191 TI - Framework for evaluating public health surveillance systems for early detection of outbreaks: recommendations from the CDC Working Group. AB - The threat of terrorism and high-profile disease outbreaks has drawn attention to public health surveillance systems for early detection of outbreaks. State and local health departments are enhancing existing surveillance systems and developing new systems to better detect outbreaks through public health surveillance. However, information is limited about the usefulness of surveillance systems for outbreak detection or the best ways to support this function. This report supplements previous guidelines for evaluating public health surveillance systems. Use of this framework is intended to improve decision-making regarding the implementation of surveillance for outbreak detection. Use of a standardized evaluation methodology, including description of system design and operation, also will enhance the exchange of information regarding methods to improve early detection of outbreaks. The framework directs particular attention to the measurement of timeliness and validity for outbreak detection. The evaluation framework is designed to support assessment and description of all surveillance approaches to early detection, whether through traditional disease reporting, specialized analytic routines for aberration detection, or surveillance using early indicators of disease outbreaks, such as syndromic surveillance. PMID- 15129192 TI - Awareness of stroke warning signs--17 states and the U.S. Virgin Islands, 2001. AB - Stroke is the third leading cause of death in the United States and a major cause of disabilities among adults. Since 1900, the number of stroke deaths has declined, and substantial advances have been made in the diagnosis and treatment of ischemic stroke during the previous decade; however, the proportion of deaths that occur before patients are transported to hospitals has increased to nearly half of all stroke deaths. One of the national health objectives for 2010 is to increase the proportion of persons who are aware of the early warning symptoms and signs of stroke (objective no. 12.8). To assess public awareness and knowledge of the proper emergency response, CDC analyzed 2001 data from the Behavioral Risk Factor Surveillance System (BRFSS) in 17 states and the U.S. Virgin Islands (USVI). This report summarizes the results of that analysis, which indicated that public awareness of several stroke signs is high, but the ability to recognize the five major warning signs is low. Education campaigns are needed to increase public awareness of stroke signs and the necessity of calling 911 when persons are suffering a possible stroke. PMID- 15129193 TI - Spina bifida and anencephaly before and after folic acid mandate--United States, 1995-1996 and 1999-2000. AB - Neural tube defects (NTDs) are serious birth defects of the spine (e.g., spina bifida) and the brain (e.g., anencephaly) that occur during early pregnancy, often before a woman knows she is pregnant; 50%-70% of these defects can be prevented if a woman consumes sufficient folic acid daily before conception and throughout the first trimester of her pregnancy. In 1992, to reduce the number of cases of spina bifida and other NTDs, the U.S. Public Health Service (USPHS) recommended that all women capable of becoming pregnant consume 400 microg of folic acid daily. Three approaches to increase folic acid consumption were cited: 1) improve dietary habits, 2) fortify foods with folic acid, and 3) use dietary supplements containing folic acid. Mandatory fortification of cereal grain products went into effect in January 1998; during October 1998-December 1999, the reported prevalence of spina bifida declined 31%, and the prevalence of anencephaly declined 16%. Other studies have indicated similar trends. To update the estimated numbers of NTD-affected pregnancies and births, CDC recently analyzed data from 23 population-based surveillance systems that include prenatal ascertainment of these birth defects. This report summarizes the results of that analysis, which indicate that the estimated number of NTD-affected pregnancies in the United States declined from 4,000 in 1995-1996 to 3,000 in 1999-2000. This decline in NTD-affected pregnancies highlights the partial success of the U.S. folic acid fortification program as a public health strategy. To reduce further the number of NTD-affected pregnancies, all women capable of becoming pregnant should follow the USPHS recommendation and consume 400 microg of folic acid every day. PMID- 15129194 TI - Lyme disease--United States, 2001-2002. AB - Lyme disease (LD) is caused by the spirochete Borrelia burgdorferi and is transmitted through the bite of Ixodes spp. ticks. CDC began LD surveillance in 1982, and the Council of State and Territorial Epidemiologists designated LD a nationally notifiable disease in 1991. This report summarizes the analysis of 40,792 cases of LD reported to CDC during 2001-2002. The results of that analysis indicate that annual LD incidence increased 40% during this period. The continued emergence of LD underscores the need for persons in areas where LD is endemic to reduce their risk for infection through integrated pest management, landscaping practices, repellent use, and prompt removal of ticks. PMID- 15129195 TI - Impact of heat waves on mortality--Rome, Italy, June-August 2003. AB - During June-August 2003, record high temperatures were reported across Europe; Italy was one of the countries most affected. To assess the impact of the summer 2003 heat waves on mortality, the Rome Local Health Authority analyzed temperature and daily mortality data for June-August 2003. This report summarizes the results of that analysis, which indicated that an estimated 1,094 excess deaths occurred during three major heat wave periods in 2003, an increase of 23% compared with the average annual number of deaths during 1995-2002. Improvements have been made in warning systems and prevention programs that target persons at high risk to reduce excess mortality during future heat waves. PMID- 15129196 TI - Degenerative changes following spinal fixation in a small animal model. AB - OBJECTIVE: The objective of this study was to evaluate changes of the lumbar vertebral column following fixation. DESIGN: Using an established small animal (rat) model of spinal fixation (hypomobility), 3 contiguous lumbar segments (L4, L5, L6) were fixed with a specially engineered vertebral fixation device. Spinal segments of control rats were compared with those of animals with 1, 4, or 8 weeks of fixation. Subgroups of these fixation animals subsequently had the fixation device removed for 1, 2, 4, 8, or 12 weeks to evaluate the effects of attempting to reestablish normal forces to the vertebral segments following hypomobility. SETTING: This Institutional Animal Care and Use Committee (IACUC) approved study was conducted in a university animal facility. ANIMALS: Eighty seven animals (23 controls animals and 64 fixation animals) were used in this study. Main Outcome Measures Outcome measures were degenerative changes of the vertebral bodies (VBs) and intervertebral disks (IVDs), zygapophysial (Z) joint osteophyte formation, and Z joint articular surface degeneration (ASD). Changes found in vertebral segments that were fixed (hypomobile) were compared with changes in adjacent nonfixed vertebral segments, and changes among fixation animals were compared with nonfixed controls. Main Results Very few degenerative changes were identified on the VBs and IVDs. Z joint changes were significant, both for osteophyte formation (analysis of variance [ANOVA], P <.0001) and ASD (ANOVA, P <.0001). Fixed segments had more degenerative changes than nonfixed segments for all Z joint parameters (ANOVA, P <.0001). Osteophyte formation and ASD were directly dependent on duration of fixation. CONCLUSIONS: These findings indicate that fixation (hypomobility) results in time-dependent degenerative changes of the Z joints. PMID- 15129197 TI - Paraspinal skin temperature patterns: an interexaminer and intraexaminer reliability study. AB - BACKGROUND: Paraspinal thermography is used by chiropractors as an aid in assessing the presence of vertebral subluxation. Few reliability studies have been carried out, with mixed results. Digital infrared scanning equipment is now available with location tracking that may enhance reproducibility. Digitized scans enable a computer-aided interpretation of thermographic patterns. OBJECTIVE: To assess the ability of examiners to reproduce thermal patterns. STUDY DESIGN: Repeated measures with 2 examiners assessing the same patient on 2 occasions. Thirty asymptomatic students served as subjects. METHODS: A TyTron C 3000 handheld thermographic scanner interfaced to a Microsoft Windows compatible personal computer was used for all recordings. Each examiner recorded 2 scans on each patient. It took an average of 3 minutes to complete all 4 scans. Data were exported to a spreadsheet for initial analysis, then SPSS was used for calculation of intraclass correlation coefficients (ICC). Since the starting and stopping points of scans were not always the same, care was taken to align scans visually, using well-distinguished peaks on the charts as guides. Scans were cropped to remove artifacts that might have occurred at the beginning and end of the scans. Intraexaminer and interexaminer ICCs were calculated. RESULTS: Skin temperatures ranged from 35.4 degrees C to 30.0 degrees C over all scans. The average temperatures changed little from the first to the last scans, indicating that subjects' overall skin temperatures were stable during the scanning procedure. Intraexaminer ICCs ranged from 0.953 to 0.984. The left and right channel data show slightly higher congruence than the Delta channel. The interexaminer reliability coefficients ranged from 0.918 to 0.975. Again, the Delta channel shows slightly less reliability, although the ICCs were quite high for all channels. CONCLUSION: Intraexaminer and interexaminer reliability of paraspinal thermal scans using the TyTron C-3000 were found to be very high, with ICC values between 0.91 and 0.98. Changes seen in thermal scans when properly done are most likely due to actual physiological changes rather than equipment error. PMID- 15129198 TI - A practice-based study of patients with acute and chronic low back pain attending primary care and chiropractic physicians: two-week to 48-month follow-up. AB - OBJECTIVE: This study reports pain and disability outcomes up to 4 years for chiropractic and medical patients with low back pain (LBP) and assesses the influence of doctor type and pain duration on clinical outcomes. DESIGN: Prospective, longitudinal, nonrandomized, practice-based, observational study. SETTING: Fifty-one chiropractic and 14 general practice community clinics. SUBJECTS: A total of 2870 acute and chronic ambulatory patients with LBP of mechanical origin. METHODS: Sixty chiropractic (DC) and 111 general practice (MD) physicians participated. Primary outcomes were pain, using a 100-point visual analogue scale (VAS), and functional disability, using the Revised Oswestry Disability Questionnaire. These were measured at baseline and 8 time points. Regression analysis compared acute and chronic DC and MD patients after correcting for baseline differences in the 4 cohorts. RESULTS: Most improvement was seen by 3 months and sustained for 1 year; exacerbation was seen thereafter. Acute patients demonstrated greater relief at all time points. A clinically important advantage for chiropractic patients was seen in chronic patients in the short-term (>10 VAS points), and both acute and chronic chiropractic patients experienced somewhat greater relief up to 1 year (P<.000). The advantage for DC care was prominent for chronic patients with leg pain below the knee (P<.001). More than 50% of chronic patients had over 50 days of pain in the third year. CONCLUSION: Study findings were consistent with systematic reviews of the efficacy of spinal manipulation for pain and disability in acute and chronic LBP. Patient choice and interdisciplinary referral should be prime considerations by physicians, policymakers, and third-party payers in identifying health services for patients with LBP. PMID- 15129199 TI - Implementing evidence-based guidelines for radiography in acute low back pain: a pilot study in a chiropractic community. AB - OBJECTIVE: To evaluate the ability of a systematic educational intervention strategy to change the plain radiography ordering behavior of chiropractors toward evidence-based practice for patients with acute low back pain (LBP). DESIGN: A quasi-experimental method was used comparing outcomes before and after the intervention with those of a control community. SETTING: Two communities in southern Ontario. DATA SOURCE: Mailed survey data on the management of acute LBP. Outcome Measures Plain radiography use rates for acute LBP based on responses to mailed surveys. RESULTS: Following the intervention, there was a 42% reduction in the self-report need for plain radiography for uncomplicated acute LBP (P <.025) and a 50% reduction for patients with acute LBP < 1 month (P <.025) in the intervention community. There was no significant change in the self-report need for plain radiography in the control community (P >.05). CONCLUSIONS: The educational intervention strategy used in this study appeared to have an effect in reducing the perceived need for plain radiography in acute LBP. PMID- 15129200 TI - Naloxone fails to antagonize initial hypoalgesic effect of a manual therapy treatment for lateral epicondylalgia. AB - BACKGROUND: Recent research has shown that Mulligan's Mobilization With Movement treatment technique for the elbow (MWM), a peripheral joint mobilization technique, produces a substantial and immediate pain relief in chronic lateral epicondylalgia (48% increase in pain-free grip strength).([1]) This hypoalgesic effect is far greater than that previously reported with spinal manual therapy treatments, prompting speculation that peripheral manual therapy treatments may differ in mechanism of action to spinal manual therapy techniques. Naloxone antagonism and tolerance studies, which employ widely accepted tests for the identification of endogenous opioid-mediated pain control mechanisms, have shown that spinal manual therapy-induced hypoalgesia does not involve an opioid mechanism. OBJECTIVE: The aim of this study was to evaluate the effect of naloxone administration on the hypoalgesic effect of MWM. METHODS: A randomized, controlled trial evaluated the effect of administering naloxone, saline, or no substance control injection on the MWM-induced hypoalgesia in 18 participants with lateral epicondylalgia. Pain-free grip strength, pressure pain threshold, thermal pain threshold, and upper limb neural tissue provocation test 2b were the outcome measures. RESULTS: The results demonstrated that the initial hypoalgesic effect of the MWM was not antagonized by naloxone, suggesting a nonopioid mechanism of action. CONCLUSIONS: The studied peripheral mobilization treatment technique appears to have a similar effect profile to previously studied spinal manual therapy techniques, suggesting a nonopioid-mediated hypoalgesia following manual therapy. PMID- 15129201 TI - Effect of a back belt on reaching postures. AB - OBJECTIVE: The present study investigated the effect of a back belt on reach actions. SUBJECTS: Sixteen undergraduate college students (8 male students, 8 female students) ranging in age from 18 to 22 years. Thirteen subjects were included in the final analysis. SETTING: The Department of Psychology at Miami University, Oxford, Ohio METHODS: Using a well-established set of procedures developed in our laboratory for studying reaching, seated adult participants reached for and retrieved an object placed at various distances from them. Reach distances included values both closer than and farther than each subject's maximum seated reach. The reach task had 2 conditions: picking up and retrieving a small block and skewering and retrieving a small bead with a needle. For each task condition, each subject either wore the belt or did not use a belt. RESULTS: Results indicate that when subjects wore the belt while reaching, they tended to have initial transition points (sitting to nonsitting) closer to their bodies than while not wearing the belt. That is, for a distant object, subjects were more likely to raise their bodies out of the chair rather than perform an extreme seated reach, possibly acting to preserve a greater margin of safety. CONCLUSIONS: The back belt consistently modified reaching postures by limiting extreme ranges of motion during a task that required enhanced stability. Furthermore, the methodology and analysis presented in this article when applied to chiropractic will allow us to begin thoughtful investigation of the effects of chiropractic adjustments on postural transitions and margin of safety. PMID- 15129202 TI - Safety of spinal manipulation in the treatment of lumbar disk herniations: a systematic review and risk assessment. AB - OBJECTIVE: To provide a qualitative systematic review of the risk of spinal manipulation in the treatment of lumbar disk herniations (LDH) and to estimate the risk of spinal manipulation causing a severe adverse reaction in a patient presenting with LDH. DATA SOURCES: Relevant case reports, review articles, surveys, and investigations regarding treatment of lumbar disk herniations with spinal manipulation and adverse effects and associated risks were found with a search of the literature. DATA SYNTHESIS: Prospective/retrospective studies and review papers were graded according to quality, and results and conclusions were tabulated. From the data published, an estimate of the risk of spinal manipulation causing a clinically worsened disk herniation or cauda equina syndrome (CES) in patients presenting with LDH was calculated. This was compared with estimates of the safety of nonsteroidal anti-inflammatory drugs (NSAIDs) and surgery in the treatment of LDH. RESULTS: An estimate of the risk of spinal manipulation causing a clinically worsened disk herniation or CES in a patient presenting with LDH is calculated from published data to be less than 1 in 3.7 million. CONCLUSION: The apparent safety of spinal manipulation, especially when compared with other "medically accepted" treatments for LDH, should stimulate its use in the conservative treatment plan of LDH. PMID- 15129203 TI - Bone metastases from breast cancer: guidelines for diagnosis. AB - OBJECTIVE: To discuss the case of a 62-year-old woman with prior history of breast cancer who later sought chiropractic care, and to present an overview of appropriate imaging procedures in such cases. CLINICAL FEATURES: The patient had a 6-month history of mild left-sided sacroiliac pain, which radiated into the left lateral thigh and leg. There was additional pain over the left upper ribs and left posterior arm, which had started insidiously. One year prior, she had undergone a lumpectomy. Current radiographs were negative for carcinoma. INTERVENTION AND OUTCOME: Cautious spinal manipulation and soft tissue procedures were used to treat her sacroiliac joints. The thoracic pain was left untreated. The patient improved over the first 7 visits but had pain return after swimming. After ceasing therapy, she still suffered from similar pain. CONCLUSIONS: It is important to assess patients who present with new bone pain after they already have a history of breast malignancy. Chiropractic physicians should take appropriate diagnostic steps to rule out suspected malignancy when there is no plain film evidence and biopsy is negative. PMID- 15129204 TI - Views on radiography use for patients with acute low back pain among chiropractors in an Ontario community. PMID- 15129205 TI - Cervical spine geometry correlated to cervical degenerative disease in a symptomatic group. PMID- 15129206 TI - Chiropractic care of a geriatric patient with an acute fracture-subluxation of the eighth thoracic vertebra. AB - OBJECTIVE: To describe the chiropractic care of a geriatric patient with complaints of midthoracic and low back pain. CLINICAL FEATURES: A 74-year-old woman sought chiropractic care with complaints of thoracic spinal pain following a fall. Palpation findings included hypertonicity and tenderness along with painful muscle spasms in the paraspinal musculature of the thoracolumbar spine. Limited range of thoracolumbar motion was found on extension and lateral flexion, most notably on right lateral flexion, with pain. Radiographic examination revealed a compression fracture at T8, in addition to spinographic listings. Signs of sprain injury were also detected at T8. INTERVENTION AND OUTCOME: The patient was cared for with contact-specific, high-velocity, low-amplitude adjustments to sites of vertebral subluxations and at the T8 fracture subluxation. The patient's response to care was positive. CONCLUSION: This case report describes the clinical features, care, and results of 1 geriatric patient with a thoracic compression fracture-subluxation treated with specific chiropractic procedures. The patient had an apparent decrease in pain as a result of the treatment. Due to the inherent limitations of a case report, it is inappropriate to generalize this outcome. PMID- 15129207 TI - Treatment of bipolar, seizure, and sleep disorders and migraine headaches utilizing a chiropractic technique. AB - OBJECTIVE: To discuss the use of an upper cervical technique in the case of a 23 year-old male patient with rapid-cycling bipolar disorder, sleep disorder, seizure disorder, neck and back pain, and migraine headaches. CLINICAL FEATURES: The patient participated in a high school track meet at age 17, landing on his head from a height of 10 ft while attempting a pole vault. Prior to the accident, no health problems were reported. Following the accident, the patient developed numerous neurological disorders. Symptoms persisted over the next 6 years, during which time the patient sought treatment from many physicians and other health care practitioners. INTERVENTION AND OUTCOME: At initial examination, evidence of a subluxation stemming from the upper cervical spine was found through thermography and radiography. Chiropractic care using an upper cervical technique was administered to correct and stabilize the patient's upper neck injury. Assessments at baseline, 2 months, and 4 months were conducted by the patient's neurologist. After 1 month of care, the patient reported an absence of seizures and manic episodes and improved sleep patterns. After 4 months of care, seizures and manic episodes remained absent and migraine headaches were reduced from 3 per week to 2 per month. After 7 months of care, the patient reported the complete absence of symptoms. Eighteen months later, the patient remains asymptomatic. CONCLUSION: The onset of the symptoms following the patient's accident, the immediate reduction in symptoms correlating with the initiation of care, and the complete absence of all symptoms within 7 months of care suggest a link between the patient's headfirst fall, the upper cervical subluxation, and his neurological conditions. Further investigation into upper cervical trauma as a contributing factor to bipolar disorder, sleep disorder, seizure disorder, and migraine headaches should be pursued. PMID- 15129208 TI - Thrombophilia in childhood. PMID- 15129210 TI - Fathering: supporting and celebrating it. PMID- 15129211 TI - Current beliefs and management strategies for treating infant colic. AB - INTRODUCTION: The purpose of this study was to describe the current beliefs about the etiology of colic as well as current management approaches used by pediatric health care providers. METHOD: An open-ended, short- answer survey concerning beliefs about the etiology and treatment of colic was conducted at major National Association of Pediatric Nurse Associates and Practitioners and American Academy of Pediatric meetings. The two groups of providers surveyed, pediatric nurse practitioners (PNPs) (n = 216) and pediatricians (n = 215), were similar in age and years of practice. RESULTS: PNPs were more likely to define colic as "excessive crying." No differences were found in the beliefs regarding etiology of colic. Pediatricians reported seeing more colicky infants per month. A strong relationship existed between etiology and primary treatment approach. Statistically significant differences were found between PNPs and pediatricians in approaches to management of colic. DISCUSSION: Colic remains a prevalent and mysterious malady, with a critical need for more evidence-based treatment protocols. Differences in approaches used by providers were found, with PNPs more likely to use behavioral and environmental approaches to treatment. PMID- 15129212 TI - The cutaneous manifestations and common mimickers of physical child abuse. AB - The cutaneous manifestations of physical child abuse are some of the most common and easily recognized forms of injury. To make an accurate assessment and diagnosis, it is important to differentiate between inflicted cutaneous injuries and mimickers of physical abuse. Likewise, an understanding of reporting guidelines helps guide practitioners in their decision making. PMID- 15129213 TI - Sleep in infants and young children: part two: common sleep problems. AB - Approximately 25% of children younger than 5 years experience some type of sleep problem. Whether the problem is acute or chronic, significant disruption to the child's sleep can occur and have a negative impact on the child and family. This article is the second in a two-part series on sleep in infants and young children. The purpose of this article is to provide fundamental information regarding common pediatric sleep problems for the clinician to use when assessing a child's sleep behaviors or addressing parental concerns. The definition, impact, and clinical evaluation of sleep problems are discussed. PMID- 15129214 TI - Coping patterns in mothers/caregivers of children with chronic feeding problems. AB - INTRODUCTION: In this study, with use of the Coping Health Inventory for Parents (CHIP), coping behaviors were examined in 35 mothers/ caregivers of children with chronic feeding problems during hospitalization for treatment of these problems. Coping in families of children with these problems has not been addressed in research. This information is beneficial in planning and implementing treatment and follow-up services. METHOD: Correlations and t tests were used to look at variables related to the mothers'/caregivers' coping patterns and to compare their frequency of usage of these patterns. RESULTS: The mothers/caregivers in this study were most likely to cope with their child's feeding problems by making active attempts to understand the problems and by doing activities with family members. Several child and family variables were significantly related to maternal coping patterns. DISCUSSION: These results suggest that it is important to provide a variety of resources to families of children with chronic feeding problems during their children's hospitalizations, including access to support and information groups. The results also suggest that it is beneficial for these families to acquire a thorough understanding of their child's medical situation because this understanding seems to help them in their ability to cope. PMID- 15129215 TI - Enrollment success in state children's health insurance program after free clinic referral. AB - INTRODUCTION: More than 11 million children remain uninsured and receive no primary health care despite the advent of the State Children's Health Insurance Program (S-CHIP). The purpose of this pilot study was to evaluate whether each child in a northeast Ohio county free pediatric clinic is referred to S-CHIP and whether the enrollment process is successful. METHOD: A record review was done to evaluate documentation indicating that such a referral was made. A phone call was made to each family for whom there was no evidence of application to S-CHIP. RESULTS: Twenty-eight referrals to Ohio's S-CHIP were documented (60%). Only 12% of families submitted an application to S-CHIP. Ten parents agreed to share their reasons for nonsubmission, the most common reason being that they thought their household income too high, followed by they did not think the program was needed. DISCUSSION: The mere existence of or referral to such a program is not adequate to ensure utilization of health care resources. Recommendations are made to increase enrollment and access to health care for low-income children. PMID- 15129216 TI - Contraception for adolescents: part one. PMID- 15129217 TI - A 4-1/2 year-old with a swollen knee. PMID- 15129218 TI - A rich source of clinical research data: medical records and telephone logs. PMID- 15129219 TI - Factor products in the treatment of hemophilia. PMID- 15129220 TI - Pediatric nurse practitioner sedation specialty role. PMID- 15129222 TI - Protection of children involved in research studies. PMID- 15129223 TI - Mitochondrial mutations in acute leukaemia. PMID- 15129224 TI - Hydroquinone modulates the GM-CSF signaling pathway in TF-1 cells. AB - Human leukemogens, including alkylating chemotherapeutic agents and benzene, enhance granulocyte-macrophage colony-stimulating factor (GM-CSF)-dependent proliferation of human CD34+ bone marrow (BM) cells. The extracellular signal regulated kinase (ERK) pathway plays an important role in GM-CSF-dependent proliferation and also has been implicated in the pathogenesis of acute myelogenous leukemia. Therefore, we investigated the effects of the benzene metabolite, hydroquinone (HQ), on alterations in the GM-CSF signaling pathway in TF-1 erythroleukemia cells and human CD34+ BM cells. HQ treatment in TF-1 cells results in a strong proliferative response that is dependent on ERK activation and GM-CSF production. HQ also induces ERK-dependent AP-1 activation with concomitant increased transcriptional activity of AP-1 reporter gene. However, the kinetics of ERK activation are different between rhGM-CSF and HQ in TF-1 cells: rhGM-CSF results in immediate activation of ERK, whereas HQ activation of ERK is delayed. Further, HQ and rhGM-CSF together produce an immediate increase in ERK phosphorylation, which is sustained for over 48 h. HQ also stimulates colony formation, AP-1 DNA binding and GM-CSF production in human CD34+ BM cells. These results suggest that HQ stimulates proliferation via activation of ERK/AP-1 and is at least partially mediated via the production of GM-CSF. PMID- 15129225 TI - Neurodevelopmental outcome at 8 months and 4 years among infants born full-term small-for-gestational-age. AB - OBJECTIVES: To examine the association between intrauterine growth restriction and neurodevelopmental outcome among full-term small-for-gestational-age (SGAT) infants at 8 months and 4 years of age. STUDY DESIGN: Growth parameters at birth and test scores on measures of neurodevelopmental function for 3922 children born SGAT were compared with those of 29,369 children born appropriately grown-for gestational-age term from similar economic backgrounds. Additional within SGAT/economic group comparisons were made for 1684 SGAT infants with symmetric undergrowth at birth and 2034 SGAT infants with asymmetric undergrowth at birth. RESULTS: Regardless of socioeconomic background, infants born SGAT were found to be at significantly increased risk for neurodevelopmental difficulties at 8 months and at 4 years of age. Few within SGAT/socioeconomic group differences in neurodevelopmental outcome appeared to be associated with specific pattern of growth restriction at birth. CONCLUSIONS: The present findings provide further evidence of the individual and public health impact of SGAT birth. PMID- 15129226 TI - Evidence for developmental hypopituitarism in ill preterm infants. AB - OBJECTIVE: The relationship between the concentrations of cortisol and T4 with outcome in the preterm infants has not been well studied. STUDY DESIGN: Mean cortisol (days 2, 4, and 6) and T4 values were correlated to gestational age, illness, and outcome in 210 infants using ANOVA. RESULTS: Cortisol significantly decreased and T4 increased across gestational age. For both hormones, higher values were found in infants on low ventilatory settings. Combined lower cortisol (mean < 5 microg/dl (138 nmol/l)) and T4 concentrations (<4 microg/dl) were found in 20/210 (9.5%) infants; 11/20 in a high-acuity group (22% of total) including 48% (12/25) of the deaths. Lower cortisol values were found in infants who died (p<0.005) in contrast to a lack of relationship with T4. CONCLUSIONS: Lower cortisol values in infants who died are consistent with the role for cortisol in survival. Combined lower cortisol and T4 concentrations in infants who failed to improve clinical status may suggest that these hormones are markers of a poor physiological state. In contrast, we suggest that these results reflect a developmental hypopituitarism, a necessary role for cortisone and T4 in successful early neonatal transition. Treatment of hypothyroidism in the setting of coexistent low cortisol concentrations (central dysfunction of the hypothalamic-pituitary axes) is known to precipitate cortisol crisis in older populations. Therefore, we caution against treatment of low neonatal thyroid concentrations until more is known about the relationship between cortisol and T4 preterm infant population. PMID- 15129227 TI - NICHD Conference on Kernicterus: Research on Prevention of Bilirubin-Induced Brain Injury and Kernicterus: Bench-to-Bedside--Diagnostic Methods and Prevention and Treatment Strategies. AB - In July 2003, the National Institute of Child Health and Human Development (NICHD) organized a consensus conference, where a group of experts were invited to review and discuss the current state of knowledge regarding neonatal hyperbilirubinemia and identify areas in which where future research should be directed. This paper summarizes the presentations addressing the current methodologies for direct and noninvasive assessments of serum total bilirubin concentrations as well as prevention and treatment strategies for the management of neonatal hyperbilirubinemia. PMID- 15129228 TI - Prevention and treatment of nosocomial sepsis in the NICU. AB - Nosocomial sepsis is a serious problem for neonates who are admitted for intensive care. It is associated with an increase in mortality, morbidity, and prolonged length of hospital stay. Thus, both the human and fiscal costs of these infections are high. Although the rate of nosocomial sepsis increases with the degree of both prematurity and low birth weight, no specific lab test has been shown to be very useful in improving our ability to predict who has a "real" blood-stream infection and, therefore, who needs to be treated with a full course of antibiotics. As a result, antibiotic use is double the rate of "proven" sepsis and we are facilitating the growth of resistant organisms in the neonatal intensive care unit. The purpose of this article is to describe simple changes in process, which when implemented, can reduce nosocomial infection rates in neonates and improve outcomes. PMID- 15129229 TI - Assessment of physician directiveness: using hypoplastic left heart syndrome as a model. AB - OBJECTIVE: To test the hypothesis that surgeons are more directive than pediatricians in discussions with parents of critically ill children. STUDY DESIGN: We designed a novel tool to assess physician directiveness in discussions with parents of children with hypoplastic left heart syndrome. We sent surveys containing the assessment tool as well as demographic questions to all attending physicians in Neonatology, Pediatric Critical Care, Pediatric Cardiology, and Congenital Cardiac Surgery at 14 of the largest pediatric cardiac surgery centers in the United States. Responses to the tool were used to determine a "directiveness score" for each physician. We then performed univariate and multivariable linear regression analyses to assess the association of physician specialty, race/ethnicity, religion, gender, and years in practice with the directiveness score. RESULTS: Responses from 138 physicians were analyzed. In univariate analyses, with neonatologists as the reference group, surgeons were most directive (regression coefficient 3.02, 95% confidence interval 0.69 to 5.35, P=0.01), followed by cardiologists (regression coefficient 1.67, 95% confidence interval 0.13 to 3.20, P=0.03). No other variable was associated with physician directiveness score. These results were similar in the multivariable analysis. CONCLUSIONS: These results suggest that physician specialty is associated with directiveness. It is unclear, however, what amount of direction parents of critically ill children prefer. PMID- 15129230 TI - Losartan, an angiotensin II (AT1) receptor antagonist, preserves cerebral blood flow in hypertensive patients with a history of stroke. AB - In patients with severe hypertension, chronic heart failure or a history of stroke, the lower limit of autoregulation of cerebral blood flow (CBF) is shifted to higher levels of blood pressure (BP) than those observed in healthy subjects. The aim of pharmacotherapy for hypertensive patients with an impaired autoregulation of CBF should be to reduce BP while preserving an appropriate CBF. In the present study, 16 hypertensive patients who had had an episode of stroke more than 4 weeks previously were administered the angiotensin II (AT1) receptor antagonist losartan at daily doses of 25-100 mg for 4 weeks. Systolic and diastolic blood pressures were recorded for 24 h using an ambulatory BP monitoring system. CBF in both hemispheres of the cerebrum and cerebellum was quantified using single photon emission tomography with N-isopropyl-p [123I]iodoamphetamine. At baseline, CBF was 29.7 +/- 6.7 ml/min/100 g in the cerebrum and 31.5 +/- 7.5 ml/min/100 g in the cerebellum. At the end of treatment, BP was lower, while CBF increased by 7.7% in the cerebrum, and remained at the baseline level in the cerebellum. Thus, CBF was preserved despite the reduction in BP. We consider the use of losartan is advantageous for hypertensive patients with a history of stroke in whom autoregulation of CBF is potentially impaired. PMID- 15129231 TI - Angiotensin II receptor antagonist has beneficial effect on hypertensive stroke patients? PMID- 15129232 TI - Arterial hypertension in Poland in 2002. AB - Arterial hypertension represents a serious medical, social and economic problem in Poland. Owing to a small number of studies concerning HT epidemiology in Poland and large differences in methodology, it is difficult to make an objective verification of the changes regarding principal parameters in our country within the last decade. Important programme for the assessment of the situation in our country is NATPOL PLUS, carried out in the year 2002 on a representative sample of 3051 adult Polish residents aged between 18 and 93 years, using the current diagnostic criteria for arterial hypertension (blood pressure readings obtained at three separate visits in cases of newly detected HT, different cuff sizes for different arm circumference, age range 18-93 years, rejection of first measurement during initial screening visit). Prevalence of HT in Poland is 29%, awareness-67%, and effectiveness of treatment-12%. This means that while HT affects about 8.4 million adult Poles, only 1 million of them get effective treatment. Moreover, as much as 8.7 million Poles have high normal blood pressure and they should apply active prevention. The awareness, detection and control of hypertension is much worse in men than in women. A series of NATPOL studies indicated over the period 1994-2002 a significant and rapid decrease in the awareness of one's own blood pressure among the adult Polish population, especially in small towns and villages, among less educated people, and in males. PMID- 15129233 TI - Genetic association between juvenile rheumatoid arthritis and polymorphism in the SH2D2A gene. AB - T-cell-specific adapter protein (TSAd) involved in the negative control of T-cell activation is encoded by the SH2D2A gene. Our recent studies indicate that homozygosity for short (ie GA(13) and GA(16)) alleles of the SH2D2A gene promoter is associated with development of multiple sclerosis. To study whether the same SH2D2A promoter polymorphism also contributes to the genetic susceptibility to develop juvenile rheumatoid arthritis (JRA), we examined 210 JRA patients and 558 healthy unrelated controls from Norway. The frequency of the short allele GA(13) was increased among the JRA patients compared to control (0.098 vs 0.05; P(n=8)=0.042). There was a significant increased frequency of HLA-DRB1(*)08 positive patients carrying two copies of 'short' alleles GA(13) and/or GA(16) compared to healthy controls (16% vs 6%; P(n=4)=0.016). Our data indicate that the 'short' alleles of the SH2D2A promoter could contribute to the genetic susceptibility to JRA. PMID- 15129234 TI - Promoter haplotypes of the interleukin-10 gene influence proliferation of peripheral blood cells in response to helminth antigen. AB - Since interleukin (IL)-10 is a key mediator of immunosuppression, and immunosuppression is considered an important element of helminth infection, we studied variants of the putative IL-10 gene promoter in 337 individuals from 130 families heavily exposed to infection by the tissue nematode Onchocerca volvulus. As shown by transmission disequilibrium tests, variants of the IL-10 promoter at positions -1082(G/A), -819(C/T), and -592(C/A) in the haplotype of ATA were significantly associated with high peripheral blood cell (PBC) proliferative responses to O. volvulus antigen (OvAg). No associations were observed using phytohemagglutinin-induced PBC proliferation or with qualitative or quantitative phenotypes of onchocerciasis or onchocerciasis-related skin disease. The findings are compatible with the hypothesis that the ATA haplotype causes a decrease in IL 10 production by OvAg-reactive type-1 regulatory T-lymphocytes, thereby alleviating the suppression of other T cells. To our knowledge, this is the first time that an influence of IL-10 promoter variants is shown on the adaptive immune response. PMID- 15129235 TI - Liver disease of uncertain cause. PMID- 15129236 TI - Single-agent high-dose melphalan followed by peripheral blood stem cell (PBSC) in lymphoma patients: an effective, and well-tolerated conditioning regimen. PMID- 15129237 TI - Carbon impacts made visible. PMID- 15129238 TI - Ethics of therapeutic cloning. PMID- 15129239 TI - Korea's stem-cell stars dogged by suspicion of ethical breach. PMID- 15129240 TI - BioShield defence programme set to fund anthrax vaccine. PMID- 15129241 TI - NASA opens its arms to robot options for saving telescope. PMID- 15129242 TI - Feathered fossils cause a flap in museums. PMID- 15129243 TI - Top job at NSF on hold until after US elections. PMID- 15129244 TI - AIDS drug price hike prompts calls for intervention. PMID- 15129245 TI - Tax change curtails UK university spin-offs. PMID- 15129246 TI - Global warming anomaly may succumb to microwave study. PMID- 15129247 TI - Fatal fruit bat virus sparks epidemics in southern Asia. PMID- 15129249 TI - Physics: the waiting game. PMID- 15129250 TI - Stem-cell research: crunch time for Korea's cloners. PMID- 15129251 TI - Multiskilled mouse rivals Renaissance rat. PMID- 15129252 TI - Colourful history of Japan's rat resources. PMID- 15129253 TI - Managing fisheries in a changing climate. PMID- 15129262 TI - Insignificance. PMID- 15129263 TI - Astronomy: dust-filled doughnuts in space. PMID- 15129264 TI - Regenerative medicine: self-help for insulin cells. PMID- 15129265 TI - Biomechanics: fast fish. PMID- 15129266 TI - Earth science: hot metal. PMID- 15129268 TI - Human genetics: an inflammatory issue. PMID- 15129269 TI - Cell biology: designer prions. PMID- 15129270 TI - Materials science: variations on a golden core. PMID- 15129272 TI - Moulting arthropod caught in the act. PMID- 15129273 TI - Adult pancreatic beta-cells are formed by self-duplication rather than stem-cell differentiation. AB - How tissues generate and maintain the correct number of cells is a fundamental problem in biology. In principle, tissue turnover can occur by the differentiation of stem cells, as is well documented for blood, skin and intestine, or by the duplication of existing differentiated cells. Recent work on adult stem cells has highlighted their potential contribution to organ maintenance and repair. However, the extent to which stem cells actually participate in these processes in vivo is not clear. Here we introduce a method for genetic lineage tracing to determine the contribution of stem cells to a tissue of interest. We focus on pancreatic beta-cells, whose postnatal origins remain controversial. Our analysis shows that pre-existing beta-cells, rather than pluripotent stem cells, are the major source of new beta-cells during adult life and after pancreatectomy in mice. These results suggest that terminally differentiated beta-cells retain a significant proliferative capacity in vivo and cast doubt on the idea that adult stem cells have a significant role in beta-cell replenishment. PMID- 15129274 TI - The central dusty torus in the active nucleus of NGC 1068. AB - Active galactic nuclei (AGNs) display many energetic phenomena--broad emission lines, X-rays, relativistic jets, radio lobes--originating from matter falling onto a supermassive black hole. It is widely accepted that orientation effects play a major role in explaining the observational appearance of AGNs. Seen from certain directions, circum-nuclear dust clouds would block our view of the central powerhouse. Indirect evidence suggests that the dust clouds form a parsec sized torus-shaped distribution. This explanation, however, remains unproved, as even the largest telescopes have not been able to resolve the dust structures. Here we report interferometric mid-infrared observations that spatially resolve these structures in the galaxy NGC 1068. The observations reveal warm (320 K) dust in a structure 2.1 parsec thick and 3.4 parsec in diameter, surrounding a smaller hot structure. As such a configuration of dust clouds would collapse in a time much shorter than the active phase of the AGN, this observation requires a continual input of kinetic energy to the cloud system from a source coexistent with the AGN. PMID- 15129275 TI - Dislocation-driven surface dynamics on solids. AB - Dislocations are line defects that bound plastically deformed regions in crystalline solids. Dislocations terminating on the surface of materials can strongly influence nanostructural and interfacial stability, mechanical properties, chemical reactions, transport phenomena, and other surface processes. While most theoretical and experimental studies have focused on dislocation motion in bulk solids under applied stress and step formation due to dislocations at surfaces during crystal growth, very little is known about the effects of dislocations on surface dynamics and morphological evolution. Here we investigate the near-equilibrium dynamics of surface-terminated dislocations using low-energy electron microscopy. We observe, in real time, the thermally driven nucleation and shape-preserving growth of spiral steps rotating at constant temperature dependent angular velocities around cores of dislocations terminating on the (111) surface of TiN in the absence of applied external stress or net mass change. We attribute this phenomenon to point-defect migration from the bulk to the surface along dislocation lines. Our results demonstrate that dislocation mediated surface roughening can occur even in the absence of deposition or evaporation, and provide fundamental insights into mechanisms controlling nanostructural stability. PMID- 15129276 TI - Polymerization within a molecular-scale stereoregular template. AB - Enzymes efficiently synthesize biopolymers by organizing monomer units within regularly structured molecular-scale spaces and exploiting weak non-covalent interactions, such as hydrogen bonds, to control the polymerization process. This 'template' approach is both attractive and challenging for synthetic polymer synthesis, where structurally regulated molecular-scale spaces could in principle provide solid-phase reaction sites for precision polymerization. Previously, free radical polymerization of methyl methacrylate in solutions containing stereoregular isotactic (it) or syndiotactic (st) poly(methyl methacrylate) (PMMA) has been shown to result in template synthesis of the opposite PMMA based on stereocomplex formation with van der Waals interactions. However, using the structure of a solid to determine the stereochemical structure of a polymer has not been satisfactorily achieved. Here we show that macromolecularly porous ultrathin films, fabricated by a single assembly step, can be used for the highly efficient stereoregular template polymerization of methacrylates through stereocomplex formation. This reaction mould accurately transfers its structural properties of stereoregularity, molecular weight and organization within the template to the new polymer. PMID- 15129277 TI - Contribution of stratospheric cooling to satellite-inferred tropospheric temperature trends. AB - From 1979 to 2001, temperatures observed globally by the mid-tropospheric channel of the satellite-borne Microwave Sounding Unit (MSU channel 2), as well as the inferred temperatures in the lower troposphere, show only small warming trends of less than 0.1 K per decade (refs 1-3). Surface temperatures based on in situ observations however, exhibit a larger warming of approximately 0.17 K per decade (refs 4, 5), and global climate models forced by combined anthropogenic and natural factors project an increase in tropospheric temperatures that is somewhat larger than the surface temperature increase. Here we show that trends in MSU channel 2 temperatures are weak because the instrument partly records stratospheric temperatures whose large cooling trend offsets the contributions of tropospheric warming. We quantify the stratospheric contribution to MSU channel 2 temperatures using MSU channel 4, which records only stratospheric temperatures. The resulting trend of reconstructed tropospheric temperatures from satellite data is physically consistent with the observed surface temperature trend. For the tropics, the tropospheric warming is approximately 1.6 times the surface warming, as expected for a moist adiabatic lapse rate. PMID- 15129278 TI - Partitioning of oxygen during core formation on the Earth and Mars. AB - Core formation on the Earth and Mars involved the physical separation of metal and silicate, most probably in deep magma oceans. Although core-formation models explain many aspects of mantle geochemistry, they have not accounted for the large differences observed between the compositions of the mantles of the Earth (approximately 8 wt% FeO) and Mars (approximately 18 wt% FeO) or the smaller mass fraction of the martian core. Here we explain these differences as a consequence of the solubility of oxygen in liquid iron-alloy increasing with increasing temperature. We assume that the Earth and Mars both accreted from oxidized chondritic material. In a terrestrial magma ocean, 1,200-2,000 km deep, high temperatures resulted in the extraction of FeO from the silicate magma ocean owing to high solubility of oxygen in the metal. Lower temperatures of a martian magma ocean resulted in little or no extraction of FeO from the mantle, which thus remains FeO-rich. The FeO extracted from the Earth's magma ocean may have contributed to chemical heterogeneities in the lowermost mantle, a FeO-rich D" layer and the light element budget of the core. PMID- 15129279 TI - Convergent evolution in mechanical design of lamnid sharks and tunas. AB - The evolution of 'thunniform' body shapes in several different groups of vertebrates, including whales, ichthyosaurs and several species of large pelagic fishes supports the view that physical and hydromechanical demands provided important selection pressures to optimize body design for locomotion during vertebrate evolution. Recognition of morphological similarities between lamnid sharks (the most well known being the great white and the mako) and tunas has led to a general expectation that they also have converged in their functional design; however, no quantitative data exist on the mechanical performance of the locomotor system in lamnid sharks. Here we examine the swimming kinematics, in vivo muscle dynamics and functional morphology of the force-transmission system in a lamnid shark, and show that the evolutionary convergence in body shape and mechanical design between the distantly related lamnids and tunas is much more than skin deep; it extends to the depths of the myotendinous architecture and the mechanical basis for propulsive movements. We demonstrate that not only have lamnids and tunas converged to a much greater extent than previously known, but they have also developed morphological and functional adaptations in their locomotor systems that are unlike virtually all other fishes. PMID- 15129280 TI - Female mating bias results in conflicting sex-specific offspring fitness. AB - Indirect-benefit models of sexual selection assert that females gain heritable offspring advantages through a mating bias for males of superior genetic quality. This has generally been tested by associating a simple morphological quality indicator (for example, bird tail length) with offspring viability. However, selection acts simultaneously on many characters, limiting the ability to detect significant associations, especially if the simple indicator is weakly correlated to male fitness. Furthermore, recent conceptual developments suggest that the benefits gained from such mating biases may be sex-specific because of sexually antagonistic genes that differentially influence male and female reproductive ability. A more suitable test of the indirect-benefit model would examine associations between an aggregate quality indicator (such as male mating success) and gender-specific adult fitness components, under the expectation that these components may trade off. Here, we show that a father's mating success in the cricket, Allonemobius socius, is positively genetically correlated with his son's mating success but negatively with his daughter's reproductive success. This provides empirical evidence that a female mating bias can result in sexually antagonistic offspring fitness. PMID- 15129281 TI - Naturalistic experience transforms sensory maps in the adult cortex of caged animals. AB - Much of what is known about the functional organization and plasticity of adult sensory cortex is derived from animals housed in standard laboratory cages. Here we report that the transfer of adult rats reared in standard laboratory cages to a naturalistic habitat modifies the functional and morphological organization of the facial whisker representation in the somatosensory 'barrel' cortex. Cortical whisker representations, visualized with repeated intrinsic signal optical imaging in the same animals, contracted by 46% after four to six weeks of exposure to the naturalistic habitat. Acute, multi-site extracellular recordings demonstrated suppressed evoked neuronal responses and smaller, sharper constituent receptive fields in the upper cortical layers (II/III), but not in the thalamic recipient layer (IV), of rats with naturalistic experience. Morphological plasticity of the layer IV barrel field was observed, but on a substantially smaller scale than the functional plasticity. Thus, transferring animals to an environment that promotes the expression of natural, innate behaviours induces a large-scale functional refinement of cortical sensory maps. PMID- 15129282 TI - Functional variation in LGALS2 confers risk of myocardial infarction and regulates lymphotoxin-alpha secretion in vitro. AB - Myocardial infarction (MI) has become one of the leading causes of death in the world. Its pathogenesis includes chronic formation of plaque inside the vessel wall of the coronary artery and acute rupture of the artery, implicating a number of inflammation-mediating molecules, such as the cytokine lymphotoxin-alpha (LTA). Functional variations in LTA are associated with susceptibility to MI. Here we show that LTA protein binds to galectin-2, a member of the galactose binding lectin family. Our case-control association study in a Japanese population showed that a single nucleotide polymorphism in LGALS2 encoding galectin-2 is significantly associated with susceptibility to MI. This genetic substitution affects the transcriptional level of galectin-2 in vitro, potentially leading to altered secretion of LTA, which would then affect the degree of inflammation; however, its relevance to other populations remains to be clarified. Smooth muscle cells and macrophages in the human atherosclerotic lesions expressed both galectin-2 and LTA. Our findings thus suggest a link between the LTA cascade and the pathogenesis of MI. PMID- 15129283 TI - Differential modulation of endotoxin responsiveness by human caspase-12 polymorphisms. AB - Caspases mediate essential key proteolytic events in inflammatory cascades and the apoptotic cell death pathway. Human caspases functionally segregate into two distinct subfamilies: those involved in cytokine maturation (caspase-1, -4 and 5) and those involved in cellular apoptosis (caspase-2, -3, -6, -7, -8, -9 and 10). Although caspase-12 is phylogenetically related to the cytokine maturation caspases, in mice it has been proposed as a mediator of apoptosis induced by endoplasmic reticulum stress including amyloid-beta cytotoxicity, suggesting that it might contribute to the pathogenesis of Alzheimer's disease. Here we show that a single nucleotide polymorphism in caspase-12 in humans results in the synthesis of either a truncated protein (Csp12-S) or a full-length caspase proenzyme (Csp12 L). The read-through single nucleotide polymorphism encoding Csp12-L is confined to populations of African descent and confers hypo-responsiveness to lipopolysaccharide-stimulated cytokine production in ex vivo whole blood, but has no significant effect on apoptotic sensitivity. In a preliminary study, we find that the frequency of the Csp12-L allele is increased in African American individuals with severe sepsis. Thus, Csp12-L attenuates the inflammatory and innate immune response to endotoxins and in doing so may constitute a risk factor for developing sepsis. PMID- 15129284 TI - Nitration of a peptide phytotoxin by bacterial nitric oxide synthase. AB - Nitric oxide (NO) is a potent intercellular signal in mammals that mediates key aspects of blood pressure, hormone release, nerve transmission and the immune response of higher organisms. Proteins homologous to full-length mammalian nitric oxide synthases (NOSs) are found in lower multicellular organisms. Recently, genome sequencing has shown that some bacteria contain genes coding for truncated NOS proteins; this is consistent with reports of NOS-like activities in bacterial extracts. Biological functions for bacterial NOSs are unknown, but have been presumed to be analogous to their role in mammals. Here we describe a gene in the plant pathogen Streptomyces turgidiscabies that encodes a NOS homologue, and we reveal its role in nitrating a dipeptide phytotoxin required for plant pathogenicity. High similarity between bacterial NOSs indicates a general function in biosynthetic nitration; thus, bacterial NOSs constitute a new class of enzymes. Here we show that the primary function of Streptomyces NOS is radically different from that of mammalian NOS. Surprisingly, mammalian NO signalling and bacterial biosynthetic nitration share an evolutionary origin. PMID- 15129285 TI - Integrating high-throughput and computational data elucidates bacterial networks. AB - The flood of high-throughput biological data has led to the expectation that computational (or in silico) models can be used to direct biological discovery, enabling biologists to reconcile heterogeneous data types, find inconsistencies and systematically generate hypotheses. Such a process is fundamentally iterative, where each iteration involves making model predictions, obtaining experimental data, reconciling the predicted outcomes with experimental ones, and using discrepancies to update the in silico model. Here we have reconstructed, on the basis of information derived from literature and databases, the first integrated genome-scale computational model of a transcriptional regulatory and metabolic network. The model accounts for 1,010 genes in Escherichia coli, including 104 regulatory genes whose products together with other stimuli regulate the expression of 479 of the 906 genes in the reconstructed metabolic network. This model is able not only to predict the outcomes of high-throughput growth phenotyping and gene expression experiments, but also to indicate knowledge gaps and identify previously unknown components and interactions in the regulatory and metabolic networks. We find that a systems biology approach that combines genome-scale experimentation and computation can systematically generate hypotheses on the basis of disparate data sources. PMID- 15129287 TI - Protein arrays: proteomics in multiplex. PMID- 15129292 TI - Swatting flies. PMID- 15129294 TI - Nuts and bolts. Passing the interview. PMID- 15129297 TI - Nutrient intake and BMI as predictors of severity of ADL disability over 1 year in homebound elders. AB - PURPOSE: Although it is well-recognized that good nutrition is essential to health, quality of life, and disease management, evidence of the role of nutrition, especially dietary intake, in the development of health-related disability among the more vulnerable older men and women in the community is largely unavailable. METHODS: We examined self-reported and performance-based measures at baseline (demographic, psychosocial and health-related factors, BMI, burden of disease, dietary intake from 3 24-hour dietary recalls, lower extremity physical performance, and activities of daily living [ADL]) and 1-year follow-up (ADL) from a randomly recruited sample of homebound elders. Structural equation modeling was used to simultaneously evaluate the direct and indirect relationships among baseline factors and severity of disability in activities of daily living (ADL) at 1-year in 253 homebound elders. results: Almost 32% of the homebound sample reported increased severity of disability over 1 year. The full structural model fit the data very well and revealed that better summary intake of calcium, vitamin D, magnesium, and phosphorus was directly linked to better lower extremity performance (LEP) and indirectly to less severity of disability at one year, after controlling for baseline disability and the interrelationships of other factors. Greater BMI was directly associated with worse LEP and indirectly with greater severity of disability. CONCLUSIONS: The findings from this study acknowledge aspects of the complex direct and indirect relationships between nutrient intake, BMI, and disability among homebound elders. These results suggest that nutrient intake and BMI may be reasonable targets for intervention with multiple functional outcomes. PMID- 15129296 TI - Zinc transporter LIVI controls epithelial-mesenchymal transition in zebrafish gastrula organizer. AB - Vertebrate gastrulation is a critical step in the establishment of body plan. During gastrulation, epithelial-mesenchymal transition (EMT) occurs. EMT is one of the central events of embryonic development, organ and tissue regeneration, and cancer metastasis. Signal transducers and activators of transcription (STATs) mediate biological actions such as cell proliferation, differentiation and survival in response to cytokines and growth factors, in a variety of biological processes. STATs are also important in EMT during gastrulation, organogenesis, wound healing and cancer progression. We previously showed that STAT3 is activated in the organizer during zebrafish gastrulation and its activity is essential for gastrulation movements. The requirement for STAT3 is cell autonomous for the anterior migration of gastrula organizer cells, and non-cell autonomous for the convergence of neighbouring cells. The molecular mechanisms of STAT's action in EMT, however, are unknown. Here we identify LIV1, a breast cancer-associated zinc transporter protein, as a downstream target of STAT3 that is essential and sufficient for STAT3's cell-autonomous role in the EMT of zebrafish gastrula organizer cells. Furthermore, we demonstrate that LIV1 is essential for the nuclear localization of zinc-finger protein Snail, a master regulator of EMT. These results establish a molecular link between STAT3, LIV1 and Snail in EMT. PMID- 15129298 TI - The impact of folic acid fortification of enriched grains on an elderly population: the New Mexico Aging Process Study. AB - Fortification of enriched grains with synthetic folic acid is a potential concern for the elderly population who is at higher risk for Vitamin B12 deficiency. Consuming excess amounts of naturally occurring folate or synthetic folic acid can precipitate a deficiency of Vitamin B12, resulting in neurological damage. The purpose of this study was to determine the increase in folate intake in an elderly population due to the fortification of enriched grains. Three-day diet records of 320 participants (average age 76.8 years) were evaluated for total folate intake from food and supplements before and after the fortification of enriched grains. There was a significant mean daily folate intake increase of 63.8 microg due to fortification (p < 0.0001), raising the intake of total folate to 359 microg (89.8% of RDA). Supplements containing folic acid were consumed by 66% of the participants, raising the average total folate intake of supplement users to 793 microg per day. Only 5 participants exceeded the UL of 1,000 microg folic acid per day, with all 5 of these individuals consuming more than 1,000 microg folic acid per day from supplements alone. Folic acid fortification of grains does not appear to have increased the risk of excess folic acid in this population. PMID- 15129299 TI - Are the equations published in literature for predicting resting metabolic rate accurate for use in the elderly? AB - PURPOSE: Equations published in literature for predicting resting metabolic rate (RMR) in older individuals were derived from studies with small samples of this age group or extrapolated from data of younger adults. The aim of the present investigation was therefore to validate various predictive equations by comparing calculated RMR with measured RMR in a large group of elderly subjects. SUBJECTS AND METHODS: RMR was measured by indirect calorimetry after an overnight fast in 225 female (age 67.7 +/- 5.7 y, BMI 26.7 +/- 3.9 kg/m2) and 130 male (age 67.4 +/ 5.4 y, BMI 26.7 +/- 3.2 kg/m2) participants of the longitudinal study on nutrition and health status in an aging population of Giessen, Germany, who were at least 60 years old. RESULTS: In females and males RMR was on average underestimated by 3.3% and 7.5% with the Schofield equation based on body weight, by 2.4% and 4.5% with the Schofield equation based on both weight and height, by 0.7% and 5.0% with the WHO equation based on body weight, and by 2.6% and 4.6% with the Harris-Benedict equation, respectively. RMR calculated with the WHO equation based on body weight and height was 1.8% higher in females and 3.9% lower in males compared to measured RMR. Regarding all predictive equations the difference between predicted and measured RMR were negatively correlated with measured RMR and were partly more pronounced in smokers and obese subjects than in non-smokers and subjects with a BMI < 30 kg/m2. CONCLUSION: At the group level all predictive equations used provide a valid estimation of RMR. However, on an individual basis estimation errors may be high. Thus in individuals RMR should be measured instead of being estimated. If measurements cannot be taken, population specific equations should be used for predicting RMR. PMID- 15129300 TI - Nutritional factors and risk of incident dementia in the PAQUID longitudinal cohort. AB - PURPOSE: The aim of this study was to present a summary of the results obtained in the PAQUID study regarding wine consumption, fish and seafood consumption and vitamin A and E plasma concentrations. METHODS: This study was carried out from the PAQUID (Personnes Agees QUID) cohort, an epidemiological study on cognitive and functional aging after 65 years in Southwestern France. The relationship between nutritional factors and risk of incident dementia was measured using logistic regression or Cox proportional hazards models on different subsamples of the cohort. RESULTS: Among 2950 initially non-demented subjects, moderate drinkers had a decreased relative risk of developing a dementia in the subsequent 8 years (RR=0.56) compared to non-drinkers. In a sub-sample of 1416 subjects, those who ate fish or seafood at least once a week had a significant reduced risk of incident dementia adjusted for age and sex (RR=0.66). Lastly, a nested casecontrol study was performed among 182 subjects. Adjusted for confounders, the risk of dementia was significantly increased for the lowest vitamin E concentration compared to the highest one (OR=2.54). DISCUSSION: These results are in agreement with a possible protective role of a diet rich in poly unsaturated fats and antioxidant components. However, confounding factors such as education and lifestyle may be involved and should be further explored. PMID- 15129301 TI - Carotenoid, tocopherol, and retinol concentrations in elderly human brain. AB - BACKGROUND: Antioxidants, such as tocopherols and carotenoids, have been implicated in the prevention of degenerative diseases. Although correlations have been made between diseases and tissue levels of antioxidants, to date there are no reports of individual carotenoid concentrations in human brain. OBJECTIVE: To measure the major carotenoids, tocopherols, and retinol in frontal and occipital regions of human brain. DESIGN: Ten samples of brain tissue from frontal lobe cortex and occipital cortex of five cadavers were examined. Sections were dissected into gray and white matter, extracted with organic solvents, and analyzed by HPLC. RESULTS: At least 16 carotenoids, 3 tocopherols, and retinol were present in human brain. Major carotenoids were identified as lutein, zeaxanthin, anhydrolutein, alpha- cryptoxanthin, beta- cryptoxanthin, alpha carotene, cis- and trans-betacarotene, and cis- and trans-lycopene. Xanthophylls (oxygenated carotenoids) accounted for 66-77% of total carotenoids in all brain regions examined. Similar to neural retina, the ratio of zeaxanthin to lutein was high and these two xanthophylls were significantly correlated (p <0.0001). The tocopherol isomers occurred in the brain over a wider range of mean concentrations (0.11-17.9 nmol/g) than either retinol (87.8 - 163.3 pmol/g) or the identified carotenoids (1.8-23.0 pmol/g). CONCLUSIONS: The frontal cortex, generally vulnerable in Alzheimer's disease, had higher concentrations of all analytes than the occipital cortex which is generally unaffected. Moreover, frontal lobes, but not occipital lobes, exhibited an age-related decline in retinol, total tocopherols, total xanthophylls and total carotenoids. The importance of these differences and the role(s) of these antioxidants in the brain remain to be determined. PMID- 15129302 TI - Roles of unsaturated fatty acids (especially omega-3 fatty acids) in the brain at various ages and during ageing. AB - Among various organs, in the brain, the fatty acids most extensively studied are omega-3 fatty acids. Alpha-linolenic acid (18:3omega3) deficiency alters the structure and function of membranes and induces minor cerebral dysfunctions, as demonstrated in animal models and subsequently in human infants. Even though the brain is materially an organ like any other, that is to say elaborated from substances present in the diet (sometimes exclusively), for long it was not accepted that food can have an influence on brain structure, and thus on its function. Lipids, and especially omega-3 fatty acids, provided the first coherent experimental demonstration of the effect of diet (nutrients) on the structure and function of the brain. In fact the brain, after adipose tissue, is the organ richest in lipids, whose only role is to participate in membrane structure. First it was shown that the differentiation and functioning of cultured brain cells requires not only alpha-linolenic acid (the major component of the omega-3, omega3 family), but also the very long omega-3 and omega-6 carbon chains (1). It was then demonstrated that alpha-linolenic acid deficiency alters the course of brain development, perturbs the composition and physicochemical properties of brain cell membranes, neurones, oligodendrocytes, and astrocytes (2). This leads to physicochemical modifications, induces biochemical and physiological perturbations, and results in neurosensory and behavioural upset (3). Consequently, the nature of polyunsaturated fatty acids (in particular omega-3) present in formula milks for infants (premature and term) conditions the visual and cerebral abilities, including intellectual. Moreover, dietary omega-3 fatty acids are certainly involved in the prevention of some aspects of cardiovascular disease (including at the level of cerebral vascularization), and in some neuropsychiatric disorders, particularly depression, as well as in dementia, notably Alzheimer's disease. Recent results have shown that dietary alpha linolenic acid deficiency induces more marked abnormalities in certain cerebral structures than in others, as the frontal cortex and pituitary gland are more severely affected. These selective lesions are accompanied by behavioural disorders more particularly affecting certain tests (habituation, adaptation to new situations). Biochemical and behavioural abnormalities are partially reversed by a dietary phospholipid supplement, especially omega-3-rich egg yolk extracts or pig brain. A dose-effect study showed that animal phospholipids are more effective than plant phospholipids to reverse the consequences of alpha-linolenic acid deficiency, partly because they provide very long preformed chains. Alpha linolenic acid deficiency decreases the perception of pleasure, by slightly altering the efficacy of sensory organs and by affecting certain cerebral structures. Age-related impairment of hearing, vision and smell is due to both decreased efficacy of the parts of the brain concerned and disorders of sensory receptors, particularly of the inner ear or retina. For example, a given level of perception of a sweet taste requires a larger quantity of sugar in subjects with alpha-linolenic acid deficiency. In view of occidental eating habits, as omega-6 fatty acid deficiency has never been observed, its impact on the brain has not been studied. In contrast, omega-9 fatty acid deficiency, specifically oleic acid deficiency, induces a reduction of this fatty acid in many tissues, except the brain (but the sciatic nerve is affected). This fatty acid is therefore not synthesized in sufficient quantities, at least during pregnancy-lactation, implying a need for dietary intake. It must be remembered that organization of the neurons is almost complete several weeks before birth, and that these neurons remain for the subject's life time. Consequently, any disturbance of these neurons, an alteration of their connections, and impaired turnover of their constituents at any stage of life, will tend to accelerate ageing. The enzymatic activities of sytivities of synthesis of long-chain polyunsaturated fatty acids from linoleic and alpha-linolenic acids are very limited in the brain: this organ therefore depends on an exogenous supply. Consequently, fatty acids that are essential for the brain are arachidonic acid and cervonic acid, derived from the diet, unless they are synthesized by the liver from linoleic acid and alpha linolenic acid. The age-related reduction of hepatic desaturase activities (which participate in the synthesis of long chains, together with elongases) can impair turnover of cerebral membranes. In many structures, especially in the frontal cortex, a reduction of cervonic and arachidonic acids is observed during ageing, predominantly associated with a reduction of phosphatidylethanolamines (mainly in the form of plasmalogens). Peroxisomal oxidation of polyunsaturated fatty acids decreases in the brain during ageing, participating in decreased turnover of membrane fatty acids, which are also less effectively protected against peroxidation by free radicals. PMID- 15129303 TI - Treatment and care of patients with chronic severe brain damage. Medical-ethical guidelines of the Swiss Academy of Medical Science. PMID- 15129304 TI - Evaluation of the European Academy for Medicine of Ageing "Teaching the Teachers" program (EAMA course II 1997- 1998). AB - The European Academy for Medicine of Ageing, founded by professors in geriatric medicine, provides a geriatric education program for European postgraduate physicians in geriatric medicine who are future academics in geriatric medicine in their countries. The course is organized for 30 participants involved in four one week residential sessions over two years. The program of each session involves 20 teachers, and includes state of the art lectures, student's lectures and working-group discussions. A first course took place in 1995-96, a second in 1997-98, a third has been accomplished in 1999-00, a fourth for 2001-02 and a fifth is ongoing for 2003--2004. The sessions are subjected to an evaluation program regarding the skills of the students, the value of the presentations and the satisfaction about the educational activities. The evaluations of the four sessions of EAMA course II (1997-1998) are presented here, with emphasis on the changes across the sessions for the whole group and for the individual students. The results are good to excellent for the main goals of the course, both by self evaluation and by peer-evaluation. The dynamic process of this European academic oriented geriatric education program attracts an increasing number of participants seeking the necessary skills and expertise to obtain academic position in geriatrics, and to meet international colleagues for further collaborative opportunities. Changes and adaptations of the following programs are fostered by the evaluation program to enhance weaker points. Accordingly, improvements in the programs and in the evaluation methods have been introduced during the third course, and are tested during the fourth and the fifth course. PMID- 15129305 TI - European Union Geriatric Medicine Society. Position statement on geriatric medicine and the provision of health care services to older people. PMID- 15129306 TI - Meeting the challenges for gerontological nursing in Europe: The European Nursing Academy for Care of Older persons (ENACO). AB - In order to support the urgent need for proactive and targeted investments in care for older persons a group of geriatric nurse specialists from different European countries convened in Leuven (Belgium) in November 2002 to launch the European Nursing Academy for Care of Older persons (ENACO). The mission of ENACO is to enhance outcomes of older persons and their caregivers through strengthening gerontological clinical nursing care, education, research and health policy within an interdisciplinary context. Specific objectives of ENACO are: 1) providing up-to-date education and training by "teaching the teachers". More specifically, the target groups are master's prepared nurses specialized in gerontology who can contribute and guide the development of gerontological/geriatric nursing care in their own country; 2) developing a core curriculum for basic gerontological nursing education in Europe and; 3) developing an European network of gerontological nursing expertise. More specifically, a web page providing opportunities for interactive communication as well as a mentoring program will be developed for nurses interested in the deepening and fine-tuning of their professional experience in care for older persons. The fact that care for older persons is high on the European agenda, the collaboration with the European Academy for Medicine of Ageing (EAMA), and other professional organizations in Europe and other parts of the world, are promising elements in the development of ENACO. PMID- 15129307 TI - Long-term results of repeat anterior anal sphincter repair. AB - PURPOSE: Anterior anal sphincter repair for obstetric trauma sometimes fails because of breakdown of the repair. The long-term results of repeating the overlapping repair are not known. METHODS: Twenty-three patients with repeat obstetric-related anterior sphincter repair had previously been assessed at a median of 20 months follow-up, at which time 13 patients (65 percent) felt 50 percent or greater improvement compared with their preoperative symptoms. Patients were reassessed at a median of 5 years (range, 48-86 months) using a questionnaire, an incontinence score, and telephone interview to determine current bowel function, continence, and restriction in activities of daily life and overall satisfaction with the results of surgery. RESULTS: Twenty-one of 23 patients (median age, 47 (range, 27-66) years) were contacted. One patient was lost to follow-up and one had died of an unrelated cause. Of 21 patients, one was fully continent and 12 more reported symptom improvement of 50 percent or more compared with preoperatively. Four were unchanged, and of the four whose symptoms had deteriorated, two had undergone further surgery for incontinence. Compared with the 20-month assessment, there was no significant change in continence scores (median, 12/20 (range, 1-20) vs. 7/20 (range, 2-19); 20 vs. 60 months), rating of improvement (median, 50 (range, 0-100) percent) at 20 and 60 months), or satisfaction (7/10 (range, 0-10) at 20 and 60 months). CONCLUSIONS: Repeat anterior sphincter repair results in improved continence for the majority of patients, with no substantial change between the short-term and long-term follow up. PMID- 15129309 TI - Vacuum-assisted closure therapy: a new treatment option for recurrent pilonidal sinus disease. Report of three cases. AB - No single treatment option available for symptomatic pilonidal disease is entirely satisfactory. In our department, we have treated successfully three cases of pilonidal disease with vacuum-assisted closure therapy. We describe our initial experience with this technique. In two cases, the pilonidal sinus was primarily excised, a split skin graft applied to cover the defect, and the vacuum assisted closure pump applied over the skin graft for a period of four days on a continuous negative pressure of 50 mmHg. The third case was treated and completely healed with vacuum-assisted closure therapy alone. To our knowledge, this is the first report in the literature describing the use of vacuum-assisted closure therapy for this condition. We propose this therapy as an alternative adjunctive treatment for pilonidal disease. PMID- 15129310 TI - Ileosigmoidal knotting: outcome in 63 patients. AB - PURPOSE: This study was designed to review the outcomes of 63 patients with ileosigmoidal knotting. METHODS: Sixty-three, surgically treated patients (47 males; 74.6 percent) were reviewed retrospectively. The mean age was 45.6 (range, 7-75) years. The most common symptoms were abdominal pain and obstipation, and the most common signs were abdominal tenderness and distention. The preoperative diagnosis was obstructive emergencies in 49 patients (77.8 percent) and nonobstructive emergencies in 14 (22.2 percent). RESULTS: All patients underwent emergency laparotomy. The most common type of ileosigmoidal knotting was Type 1A in 30 patients (47.6 percent), in which the active ileum encircled the passive sigmoid colon in a clockwise direction. Fifty patients (79.4 percent) developed gangrenous bowel. Resection of gangrenous segments and enteroenteric or enterocolic anastomosis combined with the Hartmann procedure was the most preferred operation, used in 34 patients (54 percent). The mortality rate was 15.9 percent (10 patients), and toxic shock was the most frequent cause of death. CONCLUSIONS: Ileosigmoidal knotting is a rare but serious form of intestinal obstruction. Its preoperative diagnosis is difficult and may present as an obstructive or nonobstructive emergency. Early and effective resuscitation, prompt surgical intervention selected on the basis of clinical and operative findings, and effective postoperative intensive care are the basis of treatment. PMID- 15129311 TI - Open vs. closed lateral internal sphincterotomy for idiopathic fissure-in-ano: a prospective, randomized, controlled trial. AB - PURPOSE: Internal sphincterotomy remains the "gold standard" for treatment of anal fissure but is associated with a risk of imperfect continence. Recent studies have suggested that surgical technique (open vs. closed) may influence incontinence rates after sphincterotomy. This study was designed to assess the short-term and long-term incidence of incontinence after open and closed internal sphincterotomy. METHODS: Seventy-nine patients were randomly assigned to open or closed internal sphincterotomy, performed in standardized fashion by trainee staff. Standardized questionnaires assessing continence (modified Wexner score) were administered preoperatively and at 1, 6, and 52 weeks. Postoperative stay, pain scores, complications, and fissure healing were prospectively assessed by an independent observer. RESULTS: Three patients were lost to follow-up, leaving 36 closed (16 males; mean age, 45.1 years) and 40 open (21 males; mean age, 47.9 years) internal sphincterotomy patients for assessment. All operations were performed as day case procedures with no readmissions. At six weeks postoperative, 96 percent of fissures had healed. There were no significant differences in pain scores between closed and open internal sphincterotomy at Day 1 or Day 3 postoperative. New incontinence of any grade was seen in 6.8 percent of patients at 52-week follow-up. Three patients (4.1 percent, 1 closed, 2 open) suffered major incontinence at 52 weeks. There were no significant differences in continence at 1, 6, or 52 weeks, although more open patients experienced minor imperfections at 1 week. CONCLUSIONS: Incontinence after internal sphincterotomy is not insignificant. The technique (closed vs. open) does not seem to influence incontinence rates. PMID- 15129312 TI - Acute hemorrhagic rectal ulcer syndrome: a new clinical entity? Report of 19 cases and review of the literature. AB - PURPOSE: Acute hemorrhagic rectal ulcer syndrome is characterized by sudden onset, painless, and massive hemorrhage from rectal ulcer(s) in patients with serious underlying illnesses. It is a matter of controversy whether acute hemorrhagic rectal ulcer syndrome is a distinct clinical entity. This is the first Asian report on acute hemorrhagic rectal ulcer syndrome to be made outside Japan. METHODS: From January 1989 to December 1999, 8085 patients underwent total colonoscopy at our institution. We retrospectively analyzed the medical records and colonoscopic files. The diagnosis of acute hemorrhagic rectal ulcer syndrome was made by means of the clinical, histologic, and colonoscopic findings. RESULTS: Among the 8085 patients, 19 patients (11 males; mean age, 71.2 +/- 10.1 years) were diagnosed with acute hemorrhagic rectal ulcer syndrome, which accounted for 2.8 percent of the patients with massive lower gastrointestinal bleeding. The duration from hospitalization to the onset of massive bleeding ranged from 3 to 14 (mean, 9 +/- 3.3) days. Characteristics of colonoscopic appearance were solitary or multiple rectal ulcer(s), with round, circumferential, geographical, or Dieulafoy-like lesions located within a mean of 4.7 cm +/- 1.5 cm from the dentate line. Histopathologically, the lesions appeared as necrosis with denudation of covering epithelium, hemorrhage, and multiple thrombi in the vessels of the mucosa and underlying stroma, which is considered to be similar to stress-related mucosa injury. Successful hemostasis was obtained in 74 percent (14/19) of patients with direct therapeutic maneuvers. Prognosis was largely dependent on accurate diagnosis and management of the underlying disorders. CONCLUSIONS: We assert that acute hemorrhagic rectal ulcer syndrome is a rare but important entity and stress that awareness of this clinical entity should lead to a high index of suspicion resulting in early detection, diagnosis, and appropriate therapy. PMID- 15129315 TI - HPV typing in Brazilian patients with epidermodysplasia verruciformis: high prevalence of EV-HPV 25. AB - BACKGROUND: Epidermodysplasia verruciformis is a rare genetic disorder characterized by development of lesions associated with HPV#5 or HPV#8 in early childhood; malignant transformation occurs in approximately half of individuals during adulthood. OBJECTIVE: Our goal was to study the presence and spectrum of EV-HPV types in Brazilian EV patients, a population that had never been studied in this regard. PATIENTS AND METHODS: Forty-one biopsies from different lesions (benign and skin tumors) and one biopsy from clinically normal skin from each of 20 Brazilian patients with EV were studied for HPV typing using nested PCR. RESULTS: EV-HPV DNA was detected in all 41 skin lesions of the patients and was also identified in specimens considered as normal skin from 8 patients (40%). In this study HPV-EV 25 was the most prevalent (70%), and HPV 14d (67%) was highly associated with malignant lesions. CONCLUSION: EV-HPV 25 was the most prevalent in our study. The noteworthy association of EV-HPV type 14d with skin cancers suggests its possible oncogenic role in malignant transformation in this population. PMID- 15129316 TI - Use of imiquimod cream 5% in the treatment of localized morphea. AB - Fibrosis is characterized by the increased deposition of collagen and other matrix components by fibroblasts. This process occurs as a reaction to inflammation and is mediated by numerous cytokines including transforming growth factor beta (TGF-beta). Localized cutaneous scleroderma or morphea is characterized by fibrosis. Current treatment for morphea includes topical, intralesional, or systemic corticosteroids, vitamin D analog (calcitriol and calcipotriol), photochemotherapy, laser therapy, antimalarials, phenytoin, D penicillamine, and colchicine, all with varying degrees of success. In this case report, imiquimod cream 5% (Aldara), which induces interferon and in turn inhibits TGF-beta, was employed to treat morphea. PMID- 15129317 TI - Nail psoriasis: combined therapy with systemic cyclosporin and topical calcipotriol. AB - BACKGROUND: Nail psoriasis is a common problem in psoriatic patients and often it is difficult to cure. Several treatments have been proposed in the last decade using new molecules like vitamin-D analog and/or immunosoppressive drugs both systemically and locally. OBJECTIVE: Our goal was to evaluate a combination of cyclosporin and topical calcipotriol cream versus cyclosporin alone in a matched group of patients treated with cyclosporin alone. METHOD: Fifty-four patients affected by severe psoriasis and nail involvement were selected and matched for severity of nail involvement, sex, age, and cyclosporin dosage. Group A included 21 patients treated with cyclosporin alone (3.5 mg/kg/day) for three months. Group B included 33 patients treated with the same cyclosporin dosage plus, for the same time, topical application of calcipotriol cream twice a day. Evaluation for clinical improvement was the personal feeling of the patient after three months, while clinical appearance of the lesions was evaluated by the same dermatologist using digital pictures and who was blind as to the treatment of the patient. A score ranging from + to +++ was used in order to evaluate the improvement, and data were statistically evaluated with the Wilcoxon test. RESULTS: Both cyclosporin alone and a combination of cyclosporin with topical calcipotriol twice a day were useful for treating nail psoriasis after three months of therapy although the combined therapy showed a better overall result in both mild and severe nail psoriasis. Improvement of the clinical appearance of the nail lesions was seen in about 79% of patients in group B (p < or = 0.0004) versus about 47% of patients in group A (p < or = 0.15). CONCLUSIONS: In patients with severe involvement of nail psoriasis we suggest the use of a combination of topical calcipotriol twice a day with systemic treatment such as cyclosporine. PMID- 15129318 TI - Nipple and areolar eczema in the breastfeeding woman. AB - Eczema of the nipple and areola can occasionally develop in the breastfeeding patient and is associated with sore, burning, and painful areolae and nipples. Management includes elimination of any precipitating allergens and irritant and the judicious use of appropriate topical corticosteroid preparations. Patients must also be offered effective pain control and support. The proper diagnosis and management of eczema of the nipple and areola in breastfeeding patients will prevent premature and iatrogenic weaning. PMID- 15129319 TI - Role of extracellular adenosine triphosphate in human skin. AB - BACKGROUND: The nucleotide adenosine triphosphate (ATP) has long been known to drive and participate in countless intracellular processes. Extracellular ATP and its metabolite adenosine have also been shown to exert a variety of effects on nearly every cell type in human skin. Knowledge of the sources and effects of extracellular ATP in human skin may help shape new therapies for skin injury, inflammation, and numerous other cutaneous disorders. OBJECTIVE: The objective of this review is to introduce the reader to current knowledge regarding the sources and effects of extracellular ATP in human skin and to outline areas in which further research is necessary to clarify the nature and mechanism of these effects. CONCLUSION: Extracellular ATP seems to play a direct role in triggering skin inflammatory, regenerative, and fibrotic responses to mechanical injury, an indirect role in melanocyte proliferation and apoptosis, and a complex role in Langerhans cell-directed adaptive immunity. PMID- 15129324 TI - Identification of immune-relevant genes from atlantic salmon using suppression subtractive hybridization. AB - In order to probe the interaction between an invading microorganism and its host, we have investigated differential gene expression in Atlantic salmon (Salmo salar) experimentally infected with the pathogen Aeromonas salmonicida, the causative agent of furunculosis. Subtractive cDNA libraries were constructed by suppression subtractive hybridization (SSH) from 3 immune-relevant tissues at 2 time points during the infection process. Both forward- and reverse-subtracted libraries were generated, and approximately 200 clones were sequenced from each library, giving a total of 1778 expressed sequence tags (ESTs), which were annotated according to functional categories and deposited in GenBank (BQ035314 BQ037059). Numerous genes involved in signal transduction, innate immunity, and other processes have been uncovered in the subtractive libraries. These include known acute-phase reactants, along with more novel genes encoding proteins such as tachylectin, hepcidin, precerebellin-like protein, O-methyltransferase, a putative saxitoxin-binding protein, and others. A subset of genes that were represented in the subtracted libraries was further analyzed by virtual Northern, or reverse transcription-polymerase chain reaction (RT-PCR) assays to verify their differential expression as a result of infection. PMID- 15129325 TI - Enzyme inhibitors of marine microbial origin with pharmaceutical importance. AB - Several enzyme inhibitors with various industrial uses were isolated from bacteria and actinomycetes living in the marine environment. These inhibitors are useful in medicine and agriculture. All the compounds, except the monoamine oxidase inhibitors, are novel, and their activities have been characterized. PMID- 15129326 TI - Population structure and genetic variability of six bar wrasse (Thallasoma hardwicki) in northern South China Sea revealed by mitochondrial control region sequences. AB - The genetic relationships among northern South China Sea populations of the six bar wrasse (Thallasoma hardwicki) were investigated. Fish collected from the Solomon Islands were used for geographic comparison. In 1998 and 1999, a total of 100 fish were sampled from 6 localities of the northern South China Sea and 3 localities of the Solomon Islands. Genetic variations in DNA sequences were examined from the first hypervariable region (HVR-1) of the mitochondrial control region, as amplified by polymerase chain reaction. High levels of haplotypic diversity (h = 0.944 +/- 0.0016, pi = 0.0224 +/- 0.01171) in the HVR-1 region of the mitochondrial control region of T. hardwicki were detected. This yielded 94 haplotypes that exhibited a minimum spanning tree with a starburst structure, suggestive of a very recent origin for most haplotypes. Neutrality tests indicated that the pattern of genetic variability in T. hardwicki is consistent either with genetic hitchhiking by an advantageous mutation or with population expansion. Partitioning populations into coherent geographic groups divided the northern South China Sea samples (Phi(CT) = 0.0313, P < 0.001) into 3 major groups: a north-central group composed of northwestern Taiwan and northern Vietnam; a southwestern group containing southern Vietnam; and a southern group including the central Philippines. These results are in concordance with mesoscale boundaries proposed by allozyme markers, thus highlighting the importance of identifying transboundary units for the conservation and management of fisheries in the South China Sea. PMID- 15129327 TI - Zebrafish cDNA encoding multifunctional Fatty Acid elongase involved in production of eicosapentaenoic (20:5n-3) and docosahexaenoic (22:6n-3) acids. AB - Enzymes that increase the chain length of fatty acids are essential for biosynthesis of highly unsaturated fatty acids. The gLELO gene encodes a protein involved in the elongation of polyunsaturated fatty acids in the fungus Mortierella alpina. A search of the GenBank database identified several expressed sequence tag sequences, including one obtained from zebrafish (Danio rerio), with high similarity to gLELO. The full-length transcript ZfELO, encoding a polypeptide of 291 amino acid residues, was isolated from zebrafish liver cDNA. The predicted amino acid sequence of the open reading frame shared high similarity with the elongases of Caenorhabditis elegans and human. When expressed in Saccharomyces cerevisiae, the zebrafish open reading frame conferred the ability to lengthen the chain of a range of C18, C20, and C22 polyunsaturated fatty acids, indicating not only that biosynthesis of 22:6n-3 from 18:3n-3 via a 24-carbon intermediate is feasible, but also that one elongase enzyme can perform all three elongation steps required. The zebrafish enzyme was also able to elongate monounsaturated and saturated fatty acids, and thus demonstrates a greater level of promiscuity in terms of substrate use than any elongase enzyme described previously. PMID- 15129328 TI - Fish as bioreactors: transgene expression of human coagulation factor VII in fish embryos. AB - A plasmid containing human coagulation factor VII (hFVII) complementary DNA regulated by a cytomegalovirus promoter was microinjected into fertilized eggs of zebrafish, African catfish, and tilapia. The active form of hFVll was detected in the fish embryos by various assays. This positive expression of human therapeutic protein in fish embryos demonstrates the possibility of exploitation of transgenic fish as bioreactors. PMID- 15129329 TI - Endovascular stenting for restenosis of the intracranial vertebrobasilar artery after balloon angioplasty: two case reports and review of the literature. AB - Percutaneous transluminal balloon angioplasty (PTA) has been used in the treatment of critical stenosis of the intracranial vertebrobasilar artery (VBA). PTA of the intracranial VBA carries the risk of fatal complications such as arterial dissection or acute occlusion as well as postoperative restenosis. The estimated risk of periprocedural complications and restenosis were approximately 20% and 27%. The use of recently developed stents could prevent these problems of PTA. We present two cases of restenosis of the intracranial VBA after PTA which stenoses were successfully retreated with endovascular stenting using flexible coronary stents without any complications. Neither restenosis nor other recurrent symptoms were observed during the 4- and 6-month follow-up period. Reviewing the literature of 33 cases and our 2 cases, the overall complication rates related to stenting and restenosis were 5.6% and 7.8%. Endovascular stenting for the treatment of intracranial VBA can reduce the risk of arterial dissection and restenosis. PMID- 15129330 TI - Late stenosis of aorto-aortic bypass graft treated by self-expandable stent implantation. PMID- 15129331 TI - Lower gastrointestinal bleeding from the internal iliac artery: angiographic demonstration of an iliac arteriocolic fistula. AB - A rare source of potentially massive lower gastrointestinal hemorrhage in women is advanced gynecologic malignancy. Such patients can develop gastrointestinal hemorrhage with or without prior pelvic irradiation, due to arteriocolic fistulas. Angiography permits the correct diagnosis and subsequent embolotherapy. PMID- 15129332 TI - Coil embolization treatment for perioperative pulmonary artery rupture related to Swan-Ganz catheter placement. PMID- 15129334 TI - Guidelines for stenting in infrainguinal arterial disease. PMID- 15129335 TI - Retrieving a malpositioned tracheobronchial stent utilizing Amplatz gooseneck snare: a case report. AB - Tracheobronchial stents play a major role in the management of tracheobronchial stenoses and/or bronchomalacia. A very few techniques have been described in literature to remove misplaced or malpositioned tracheobronchial stents. This is a case report describing a technique to retrieve a malpositioned stent utilizing Amplatz gooseneck snare. PMID- 15129336 TI - CT-guided transthoracic core biopsy for pulmonary tuberculosis: diagnostic value of the histopathological findings in the specimen. AB - We evaluated the value of CT-guided transthoracic core biopsy for the diagnosis of mycobacterial pulmonary nodules. The 30 subjects in this study had pulmonary nodules that had been either diagnosed histopathologically as tuberculosis or were suspected as tuberculosis based on a specimen obtained by CT-guided transthoracic core biopsy. The histopathological findings, the existence of acid fast bacilli in the biopsy specimens, and the clinical course of the patients after the biopsy were reviewed retrospectively. Two of the three histological findings for tuberculosis that included epithelioid cells, multinucleated giant cells and caseous necrosis were observed in 21 of the nodules which were therefore diagnosed as histological tuberculosis. Six of these 21 nodules were positive for acid-fast bacilli, confirming the diagnosis of tuberculosis. Thirteen of the 21 nodules did not contain acid-fast bacilli but decreased in size in response to antituberculous treatment and were therefore diagnosed as clinical tuberculosis. Seven nodules with only caseous necrosis were diagnosed as suspected tuberculosis, with a final diagnosis of tuberculosis being made in 4 of the nodules and a diagnosis of old tuberculosis in 2 nodules. Two nodules with only multinucleated giant cells were diagnosed as suspected tuberculosis with 1 of these nodules being diagnosed finally as tuberculosis and the other nodule as a nonspecific granuloma. When any two of the three following histopathological findings--epithelioid cells, multinucleated giant cells or caseous necrosis--are observed in a specimen obtained by CT-guided transthoracic core biopsy, the diagnosis of tuberculosis can be established without the detection of acid-fast bacilli or Mycobacterium tuberculosis. PMID- 15129337 TI - Hypogastric arterial selective and superselective embolization for severe postpartum hemorrhage: a retrospective review of 36 cases. AB - We report on embolization in 36 cases of postpartum hemorrhage (PPH). The 36 patients with severe PPH, including one patient who had undergone an emergency hysterectomy, were transferred to the regional interventional vascular radiology unit in a mean time of 6 hours 12 min. Bilateral occlusion of the anterior trunk of the hypogastric arteries was carried out using gelatin sponge. Immediate success was achieved in all cases. In 3 cases, however, a second embolization was necessary before day 2. In 17%, complementary nonvascular surgery was performed. Complications included one puncture site false aneurysm treated by compression, two cases of regressive lower limb paraesthesia, one femoral vein thrombosis, and nonsignificant puncture site hematomas (19.5%). Long-term follow-up was conducted in 23 patients: 91% resumed regular menstrual cycles, 8.7% dysmenorrhea. New pregnancy occurred in 13% (two full-term pregnancies and one voluntary termination). Immediate efficacy, low morbidity and preservation of fertility make embolization the technique of choice for severe PPH. PMID- 15129338 TI - Acute ischemic sialadenitis following facial artery embolization. PMID- 15129339 TI - Endovascular stent graft: treatment of pseudoaneurysm of the superior mesenteric artery. PMID- 15129341 TI - Administration of iodized oil resulted in impaired liver function due to enhanced portosystemic shunting. PMID- 15129342 TI - Mechanical thrombectomy of iliocaval thrombosis using a protective expandable sheath. AB - We report a case of successful percutaneous treatment of a subacute ilio-caval venous thrombosis in a 64-year-old female patient by using a novel combination of a rotatory fragmentation device (percutaneous thrombectomy device: PTD) and large wire basket (temporary Gunther basket filter) under temporary caval filter protection using an expandable sheath. Because the patient had multiple myeloma with increased risk for contrast media-induced renal failure, the therapeutic angiographic procedure was performed without iodinated contrast medium. Non contrast-enhanced MR venography (high-resolution True FISP) confirmed the effective thrombus removal by the percutaneous mechanical thrombectomy procedure. PMID- 15129343 TI - Intrahepatic ductal stone formation after percutaneous ethanol injection for residual hepatocellular carcinoma in the portal vein. PMID- 15129344 TI - Reperfusion of splanchnic artery aneurysm following transcatheter embolization: treatment with percutaneous thrombin injection. AB - We describe a case of reperfusion of an aneurysm of the pancreaticoduodenal artery following transcatheter coil embolotherapy. The lesion was successfully treated by direct puncture of the aneurysm under computed tomographic guidance, followed by injection of thrombin. This technique is useful when an endovascular approach is not feasible. PMID- 15129345 TI - Cutting balloon-assisted angioplasty of an anastomotic carotid-brachial bypass graft stenosis. AB - Neointimal hyperplasia leads to anastomotic stenosis in bypass grafts. These stenoses are often resistant to conventional balloon dilatation. We present a case of a carotid-brachial bypass graft stenosis, which was treated by a 5-mm cutting balloon angioplasty with a good angiographic and clinical result. PMID- 15129346 TI - Feasibility of respiratory triggering for MR-guided microwave ablation of liver tumors under general anesthesia. AB - We obtained clear and reproducible MR fluoroscopic images and temperature maps for MR image-guided microwave ablation of liver tumors under general anesthesia without suspending the artificial ventilation. Respiratory information was directly obtained from air-way pressure without a sensor on the chest wall. The trigger signal started scanning of one whole image with a spoiled gradient echo sequence. The delay time before the start of scanning was adjusted to acquire the data corresponding to the k-space center at the maximal expiratory phase. The triggered images were apparently clearer than the nontriggered ones and the location of the liver was consistent, which made targeting of the tumor easy. MR temperature images, which were highly susceptible to the movement of the liver, during microwave ablation using a proton resonance frequency method, could be obtained without suspending the artificial ventilation. Respiratory triggering technique was found to be useful for MR fluoroscopic images and MR temperature monitoring in MR-guided microwave ablation of liver tumors under general anesthesia. PMID- 15129362 TI - Communication in ecosystem management: a case study of cross-disciplinary integration in the assessment phase of the interior Columbia Basin Ecosystem Management Project. AB - Effective communication is essential to the success of collaborative ecosystem management projects. In this paper, we investigated the dynamics of the Interior Columbia Basin Ecosystem Management Project's (ICBEMP) cross-disciplinary integration process in the assessment phase. Using a case study research design, we captured the rich trail of experience through conducting in-depth interviews and collecting information from internal and public documents, videos, and meetings related to the ICBEMP. Coding and analysis was facilitated by a qualitative analysis software, NVivo. Results include the range of internal perspectives on barriers and facilitators of cross-disciplinary integration in the Science Integration Team (SIT). These are arrayed in terms of discipline based differences, organizational structures and activities, individual traits of scientists, and previous working relationships. The ICBEMP organization included a team of communication staffs (CT), and the data described the CT as a mixed group in terms of qualifications and educational backgrounds that played a major role in communication with actors external to the ICBEMP organization but a minor one in terms of internal communication. The data indicated that the CT-SIT communication was influenced by characteristics of actors and structures related to organizations and their cultures. We conclude that the ICBEMP members may not have had a sufficient level of shared understanding of central domains, such as the task at hand and ways and timing of information sharing. The paper concludes by suggesting that future ecosystem management assessment teams use qualified communications specialists to design and monitor the development of shared cognition among organization members in order to improve the effectiveness of communication and cross-disciplinary integration. PMID- 15129363 TI - Calf augmentation with supraperiostic solid prosthesis associated with fasciotomies. AB - This article describes, for the first time, the supraperiostic technique used for aesthetic purposes to remedy hemiatrophy, genu valgum, poliomyelitis sequela, and thin legs. For genu valgum and thin legs, a solid prosthesis was used, whereas for severe hemiatrophy a double light prosthesis was used as a tissue expansor, with excellent reports. The advantages of this technique are delineated further in this article. PMID- 15129364 TI - One-stage reconstruction of an alar defect using a bilobed nasolabial-nasal tip flap based on the aesthetic subunits in Orientals: case report. AB - The ala of the nose is difficult to repair for a variety of reasons, including the special texture of the skin, its shape, and the free margin. A method is reported for the one-stage reconstruction of a lateral nasal defect, including a full-thickness defect of the ala, using a bilobed flap composed of nasolabial and nasal tip skin. The flap was designed according to the aesthetic subunits principle, as modified for Orientals by Yotsuyanagi and colleagues in 2000. Satisfactory alar reconstruction was achieved, and good aesthetic and functional results were obtained. This flap is useful for restoring an alar defect. PMID- 15129365 TI - Abnormal aortic arch. PMID- 15129366 TI - Body weight change since menopause and percentage body fat mass are predictors of subsequent bone mineral density change of the proximal femur in women aged 75 years and older: results of a 5 year prospective study. AB - Few studies have evaluated risk factors for bone loss in elderly women. We examined risk factors associated with a 5-year longitudinal change in bone mineral density (BMD) at the hip in healthy women aged 75 years and older. The BMD of 276 women from the French EPIDOS (Epidemiologie des Osteoporoses) study was assessed in Montpellier from 1992 to 1993 and again from 1997 to 1998. BMD was measured at the femoral neck, trochanter, and Ward's area using the same Lunar densitometer. We examined the relationship between clinical and behavioral factors at baseline and their variations during follow-up, with percentage BMD change adjusted for baseline BMD. Depending on the femur subregion studied, a significant decrease in BMD (exceeding the least significant difference, i.e., > 2.8 CV) was observed in 36.2% to 51.1% of women. Multivariate analysis showed that both postmenopausal weight change before baseline and baseline percentage of fat mass were positively correlated with BMD change at the Ward's triangle and the trochanter. Yearly absolute and relative weight changes over the follow-up period were significantly associated with change of trochanter and femoral neck BMD. Our results show that maintenance of body weight throughout the postmenopause period and body fat mass play protective roles against bone loss at the proximal femur in women aged 75 years and older and suggest the value in including assessment of weight change throughout postmenopause and percentage body fat mass in screening programs for elderly women who are at higher risk of accelerated bone loss. PMID- 15129367 TI - Relationship between computed tomographic image analysis and histomorphometry for microarchitectural characterization of human calcaneus. AB - The present study aimed to characterize the relationships between several variables reflecting bone microarchitecture assessed by both computed tomographic (CT) image analysis and histomorphometry (conventional CT system) at the calcaneus. A total of 24 cadaveric specimens were studied. The mean age at death was 78 +/- 10 years (range, 53-93 years). A total of 15 sagittal sections (1 mm in width and spaced 2 mm apart) were selected for CT analysis; 6 undecalcified sections (7 microm) were analyzed for histomorphometry. The histomorphometric analysis was performed on a Leica Quantimet Q570 image analyzer. Features measured by both methods were: bone volume/tissue volume (BV/TV), trabecular thickness (Tb.Th), trabecular separation (Tb.Sp), trabecular number (Tb.N), interconnectivity index (ICI), number of nodes (N Nd), number of terminus (N Tm), node-to-node strut count (NNS), node-to-terminus strut count (NTS), terminus-to terminus strut count (TTS), marrow space star volume (SV), Euler number (EN), and fractal dimension (FD). The coefficient of correlations' values (simple linear regression) between histomorphometry and CT image analysis varied according to the parameters selected. R values were high for BV/TV, Tb.N, and Tb.Sp (range, 0.69-0.90; P < 0.01). R values were less significant for some variables also obtained from the binary image: SV (0.5, P < 0.05) and EN (0.43, P < 0.05). Finally R values were also significant for (two) variables obtained from skeletonized images, i.e., N Nd (0.4, P < 0.05) and N Tm (0.61, P < 0.01). Other correlations were not statistically significant. Moreover, for some variables the relationships between the two methods (CT analysis and histomorphometry) seemed best-described by using nonlinear models. For example, a logarithmic model was more appropriate for SV (r = 0.71, P < 0.01), N Nd (r = 0.52, P < 0.01). Finally the relationship between apparent (App) N Tm and N Tm was most satisfying when using an exponential model (r = 0.64, P < 0.01). In conclusion, trabecular bone structure measures determined on CT images show highly significant correlations with those determined using histomorphometry. The level of correlation varies according to the type of method used for characterizing bone structure, however, and the strongest correlations were found for the most basic features (Parfitt's parameters). Finally, for some variables, nonlinear models seem more appropriate. PMID- 15129368 TI - The interleukin-6/soluble interleukin-6 receptor system induces parathyroid hormone-related protein in human osteoblastic cells. AB - In the present in vitro study, we tested the hypothesis that parathyroid hormone related protein (PTHrP) might be a mediator of interleukin-6 (IL-6) and its soluble receptor (IL-6sR) in osteoblasts. We found that IL-6, within 1-20 ng/mL, added together with IL-6sR (100 ng/mL), rapidly (1 hour) increased PTHrP mRNA in human osteoblastic osteosarcoma MG-63 cells and human osteoblastic (hOB) cells from trabecular bone. PD098059, a mitogen-activated protein kinase (MAPK) kinase inhibitor, at 10 microM, and two inhibitors of protein prenylation and thus Ras activation, simvastatin (1 microM) and a farnesyltransferase (FTase) inhibitor (100 nM), but not the phosphatidylinositol 3-kinase inhibitor wortmannin, blocked the IL-6/IL-6sR-induced PTHrP expression in these cells. In addition, PD098059 as well as simvastatin and the FTase inhibitor abolished alkaline phosphatase activity and/or osteocalcin mRNA induction by the IL-6/IL-6sR in these cells. Our results support the role of the Ras/MAPK pathway as a major mechanism in the modulation of both PTHrP expression and differentiation in human osteoblasts. PMID- 15129369 TI - Two single nucleotide polymorphisms in the CYP17 and COMT Genes--relation to bone mass and longitudinal bone changes in postmenopausal women with or without hormone replacement therapy. The Danish Osteoporosis Prevention Study. AB - Sex steroids are important physiologic regulators of bone mass, and genes regulating sex steroid production and metabolism are obvious as candidate genes for osteoporosis susceptibility. We present data from a study of 1795 recent postmenopausal women, assigned to either hormone replacement therapy (HRT) or no treatment and followed for 5 years. The association between bone mass measurements and two single nucleotide polymorphisms, a T (A1) to C (A2) transition in the 5'-UTR of the cytochrome P450c17alpha (CYP17) gene and a G (Val) to A (Met) transition in exon 4 of the catechol- O-methyltransferase (COMT) gene, was evaluated. Association with CYP17 genotype was modified by body mass index (BMI). In lean women, individuals homozygous for the CYP17 A2 allele were 1 cm shorter and had lower baseline BMD (bone mineral density), BMC, and CSA (cross sectional area) in the spine and femoral neck than did other women (BMD spine A2A2: 0.975 g/cm2 versus 1.011 g/cm2 in A1A1 + A1A2, P = 0.002). Conversely, an adverse association with A2A2 and bone loss over 5 years seemed present only in overweight women, but differences were small. Response to HRT was not dependent on CYP17 genotype. COMT genotype was not associated with bone mass at baseline, bone loss in untreated women, or response to HRT. In conclusion, the A2 allele of the CYP17 T(27)-C polymorphism is associated with reduced bone mass and bone size in lean perimenopausal women, whereas high BMI protects against this negative association. The COMT G(1947)-A polymorphism is not associated with bone parameters in this study. PMID- 15129370 TI - Role of hormonal changes in the pathogenesis of osteoporosis in men. PMID- 15129371 TI - The use of M&M Rounds in departments of FM. PMID- 15129372 TI - Update on family medicine in Turkey. PMID- 15129373 TI - The future is now. PMID- 15129374 TI - Professionalism: Part I--Introduction and being a role model. PMID- 15129375 TI - The RAFT approach to academic detailing with preceptors. AB - We have adapted "academic detailing" as a method to deliver real-time faculty development to nearly all community preceptors, both rural and urban, using the RAFT technique (Rapport-building, Assessment, Focused preceptor development, Thanks/Trinkets). The method is expensive in terms of faculty time and can be difficult to schedule. However, it has proven to be a valuable, learner-centered method that reaches preceptors missed by traditional faculty development workshops. Preceptors experience support, both student and preceptor problems are identified and addressed early, and academic faculty gain a better understanding of the preceptor environment. PMID- 15129376 TI - A stroke and its consequences. PMID- 15129377 TI - Winslow Homer's "Breezing up:" Inspiring providers of community primary care. PMID- 15129378 TI - The combined effect of transferrin saturation and low density lipoprotein on mortality. AB - BACKGROUND AND OBJECTIVES: Evidence suggests that cardiovascular disease (CVD) is accelerated by the oxidation of low-density lipoprotein (LDL) in the presence of iron. This study examined whether adults with elevated iron, as measured by transferrin saturation (TS), and elevated LDL are at an increased risk for mortality. METHODS: This is a cohort study of the adult US population using the National Health and Nutrition Examination Survey 1976-1980 (NHANES II) merged with the NHANES II Mortality Study in 1992. Multivariate Cox regression was performed to determine hazard ratios (HR) for CVD and all-cause mortality for high (>55%) or low (<55%) levels of TS and high (>160 mg/dl) or low (<160 mg/dl) levels of LDL. RESULTS: An elevated LDL alone did not significantly increase CVD mortality or all-cause mortality in the adjusted model. Individuals with elevated LDL and elevated TS had a statistically significant increase in both CVD mortality and all-cause mortality (HR=5.74 and 3.53, respectively) compared to the low LDL and low TS group. CONCLUSIONS: The results of this study indicate an increased risk associated with the combination of elevated LDL and elevated TS, which suggests that iron-mediated oxidation of LDL may be a significant factor in the progression of CVD. PMID- 15129379 TI - The Clinical Hand: A curricular map for relationship-centered care. AB - Learning and teaching the complex craft of relationship-centered care in the context of the competing demands of clinical family medicine can be challenging. The "Clinical Hand" is an educational aid, which serves as a curricular map specifying what content and skills are important for relationship centered care. The Clinical Hand illuminates seven features of the clinical encounter. "Opening the Hand" symbolizes the importance of relationship and healing intention. The "Grip of Power" highlights the significance of locating, owning, aiming, and sharing power in the relationship. The "Wrist Lines of Guidance" name three goals for every visit and three types of clinical encounters. The "Fingers of Direction" identify five sequential tasks for each encounter, and the "Nails for Trouble" remind residents of the BATHE technique. The "Palm of Hope" represents a diagnostic and management tool, and a "Swinging Cultural Ape" emphasizes the importance of evolution, culture, and the need to "keep swinging." The use of the Clinical Hand in a residency program is briefly discussed. PMID- 15129380 TI - Medical students' perceptions of rural practice following a rural clerkship. AB - BACKGROUND AND OBJECTIVES: Evidence suggests that rural experiences can positively influence students' preferences for rural practice. This study examined changes in students' perceptions toward rural primary care following a required rural clerkship. METHODS: Third-year students completed pre- and post clerkship questionnaire items assessing their beliefs about primary care physicians who practice in rural communities in comparison with their urban/suburban counterparts. A factor analysis was performed, and pre- and post clerkship scale means were calculated to determine differences. RESULTS: A total of 428 (88%) students completed these questionnaires. There was a significant increase in students' perceptions of rural primary care physicians' primary care service features and medical expertise. Students perceived the physicians' work demands more positively, and there was no change in students' perceptions of the physicians' income potential. CONCLUSIONS: Results suggest that the rural primary care clerkship positively influenced students' perceptions toward rural primary care. PMID- 15129381 TI - A controlled trial of an advanced access appointment system in a residency family medicine center. AB - BACKGROUND AND OBJECTIVES: The implementation of advanced access appointment systems has improved continuity of care, patient and physician satisfaction, physician productivity, and average physician panel size in private practice and group-model HMO settings. This study's purpose was to document the patient care benefits, practice management benefits, and educational outcomes from the controlled implementation of an advanced access appointment system in a residency family medicine center. METHODS: Two faculty-resident teams were created. One team adopted the advanced access system while the other team continued using a traditional access system. Outcome measures included length of time needed to obtain an appointment (days to third available appointment), continuity (percentage of visits with the patient's designated provider), no-show rates, productivity, visits lost to outside providers, panel sizes, and patient satisfaction. Outcomes were measured at baseline and quarterly for 1 year after initial implementation. RESULTS: After implementation, the "days to third available appointment" for the advanced access group was 5 days, compared to 21 days for the traditional access group. A significant improvement in continuity (ie, a match between the primary care physician and patient) for the advanced access team was found. Comparison of no-show rates between the advanced access and traditional access teams revealed significant between-subjects effect, but controlling for within-subject variation using repeated measures ANOVA eliminated this effect. Advanced access residents increased their continuity above 50% while increasing provider satisfaction with office practice and scope of practice. CONCLUSIONS: Faculty and residents can successfully use advanced access. Advanced access can enhance residency education by reducing appointment delays and significantly increasing the patient-primary care physician match. PMID- 15129382 TI - Health care experiences of African American teen women in eastern North Carolina. AB - BACKGROUND AND OBJECTIVES: Women, especially women of color, are a vulnerable population in eastern North Carolina. This study investigated the health care experiences of Southern, rural, African American adolescent women. METHODS: Four focus groups were conducted in three counties in eastern North Carolina in the spring of 2000. A total of 44 teen women ages 16-19 participated; 38 were African American. RESULTS: Participants emphasized the following themes: (1) seeking help for sexual health, (2) obtaining health information from sources other than the doctor, (3) gender and race preferences, (4) communication with physicians, and (5) good and bad doctors' visits. CONCLUSIONS: To provide quality care to African American teenage women, it is critical to listen and understand their experiences. Most importantly, medical educators must be cognizant of cultural issues and how they can impede as well as enhance patient care. This study supports the need to educate students, residents, and practicing physicians on how to provide culturally sensitive and competent care. PMID- 15129383 TI - An international consultation: the development of family medicine in Vietnam. AB - An effective international consultation on health system reform can be approached using the five-step process of establishing goals, conducting a needs assessment, defining objectives, developing methods, and designing evaluation strategies. This structure provided guidance to a consultation we provided to the Ministry of Health, Socialist Republic of Vietnam (SRV) to review its current health care delivery system. The consultation examined all levels of health care delivery and medical education. The SRV has an extensive, but poorly staffed, "commune health center" system. There is a widespread perception that the quality of medical care is low in these health centers. People leave their communities to obtain health care elsewhere at more-specialized levels and more-expensive sites. Our consultation included an analysis of the potential effect of creating a primary health care delivery system based on the model of family medicine. In addition to consulting, part of the time spent in Vietnam was used to advocate for changes in the system to allow for movement toward a primary health care delivery system. The consultation culminated in the creation of the specialty of family medicine and in the establishment of the medical education system to train family physicians. PMID- 15129384 TI - What do African American teen women want in health care? PMID- 15129385 TI - Myth-information about family medicine: is fiction better than truth? PMID- 15129386 TI - Is it time for a 4-year family medicine residency? PMID- 15129391 TI - Intramuscular neuromuscular electric stimulation for poststroke shoulder pain: a multicenter randomized clinical trial. AB - OBJECTIVE: To assess the effectiveness of intramuscular neuromuscular electric stimulation (NMES) in reducing poststroke shoulder pain. DESIGN: Multicenter, single-blinded, randomized clinical trial. SETTING: Ambulatory centers of 7 academic rehabilitation centers in the United States. PARTICIPANTS: Volunteer sample of 61 chronic stroke survivors with shoulder pain and subluxation. INTERVENTION: Treatment subjects received intramuscular NMES to the supraspinatus, posterior deltoid, middle deltoid, and trapezius for 6 hours a day for 6 weeks. Control subjects were treated with a cuff-type sling for 6 weeks. Main outcome measure Brief Pain Inventory question 12 (BPI 12), an 11-point numeric rating scale administered in a blinded manner at the end of treatment, and at 3 and 6 months posttreatment. RESULTS: The NMES group exhibited significantly higher proportions of success based on the 3-point or more reduction in BPI 12 success criterion at the end of treatment (65.6% vs 24.1%, P<.01), at 3 months (59.4% vs 20.7%, P<.01), and at 6 months (59.4% vs 27.6%, P<.05). By using the most stringent "no pain" criterion, the NMES group also exhibited significantly higher proportions of success at the end of treatment (34.4% vs 3.4%, P<.01), at 3 months (34.4% vs 0.0%, P<.001), and at 6 months (34.4% vs 10.3%, P<.05). CONCLUSIONS: Intramuscular NMES reduces poststroke shoulder pain among those with shoulder subluxation and the effect is maintained for at least 6 months posttreatment. PMID- 15129392 TI - Botulinum toxin type B in upper-limb poststroke spasticity: a double-blind, placebo-controlled trial. AB - OBJECTIVE: To determine whether botulinum toxin type B (BTX-B) is effective in controlling upper-limb spasticity. DESIGN: A single-site, double-blind, placebo controlled, randomized trial and open-label study. SETTING: Outpatient. PARTICIPANTS: Subjects with an Ashworth Scale score of 2 or more at the elbow, wrist, and fingers. INTERVENTIONS: Subjects were injected with 10000 U of BTX-B or placebo at the elbow, wrist, and finger flexors. Main outcome measures Measures recorded at weeks 0, 2, 4, 8, 12, and 16, with a 12-week open-label study. Ashworth Scale score, a global assessment of change (GAC), adverse events and mouse neutralization antibody testing. RESULTS: BTX-B did not decrease muscle tone in the elbow, wrist, or finger flexors at 10000 U over the 16-week period. A decrease in Ashworth Scale score for the BTX-B patient group was present at the wrist at week 2 of the double-blind study (P=.003) but was not statistically significant at other visits. In the open-label study, improvement was noted at week 4 for the elbow (P=.039), wrist (P=.002), finger (P=.001), and thumb flexors (P=.002). In the double-blind study, the Physician GAC did not reach significance. Dry mouth was reported by 8 of 9 BTX-B subjects in the double-blind study. Mouse neutralization antibodies were negative. CONCLUSIONS: Our study does not show a significant decrease in tone from 10000 U of BTX-B. Dry mouth was common. PMID- 15129393 TI - A randomized controlled trial of weight-bearing versus non-weight-bearing exercise for improving physical ability after usual care for hip fracture. AB - OBJECTIVE: To compare the effects of weight-bearing and non-weight-bearing home exercise programs and a control program on physical ability (strength, balance, gait, functional performance) in older people who have had a hip fracture. DESIGN: Randomized controlled trial with 4-month follow-up. SETTING: Australian community-dwellers (82%) and residents of aged care facilities who had completed usual care after a fall-related hip fracture. PARTICIPANTS: One hundred twenty older people entered the trial, 40 per group (average age +/- standard deviation, 79+/-9y) and 90% completed the 4-month retest. INTERVENTION: Home exercise prescribed by a physical therapist. MAIN OUTCOME MEASURES: Strength, balance, gait, and functional performance. RESULTS: At the 4-month retest, there were differences between the groups in the extent of improvement since the initial assessment for balance (F(10,196)=2.82, P<.001) and functional performance (F(6,200)=3.57, P<.001), but not for strength (F(12,190)=1.09, P=.37) or gait (F(8,200)=.39, P=.92). The weight-bearing exercise group showed the greatest improvements in measures of balance and functional performance (between-group differences of 30%-40% of initial values). CONCLUSIONS: A weight-bearing home exercise program can improve balance and functional ability to a greater extent than a non-weight-bearing program or no intervention among older people who have completed usual care after a fall-related hip fracture. PMID- 15129394 TI - A randomized, prospective study of the effects of Tai Chi Chun exercise on bone mineral density in postmenopausal women. AB - OBJECTIVE: To evaluate the potential benefits of programmed Tai Chi Chun (TCC) exercise on the weight-bearing bones of early postmenopausal women. DESIGN: Age matched and randomized prospective intervention. SETTING: University medical school. PARTICIPANTS: One hundred thirty-two healthy postmenopausal women (mean age, 54.0+/-3.5y) within 10 years of menopause onset were recruited and randomized into the TCC exercise group (n=67) or the sedentary control group (n=65). INTERVENTION: Supervised TCC exercise was performed by the TCC group for 45 minutes a day, 5 days a week, for 12 months; control subjects retained a sedentary life style. Main outcome measures Bone mineral density (BMD) was measured in the lumbar spine and proximal femur by using dual-energy x-ray absorptiometry and in the distal tibia by using multislice peripheral quantitative computed tomography (pQCT). All BMD measurements were repeated after 12 months in both groups. Fracture rate was also documented. RESULTS: Baseline measurements showed homogeneity in age, anthropometric variables, and menstruation status between the TCC and control groups. Exactly 81.6% of the subjects in the TCC group and 83.1% of subjects in the control group completed the 12-month follow-up study. BMD measurements revealed a general bone loss in both TCC and sedentary control subjects at all measured skeletal sites, but with a reportedly slower rate in the TCC group. A significant 2.6- to 3.6-fold retardation of bone loss (P<.01) was found in both trabecular and cortical compartments of the distal tibia in the TCC group as compared with the controls, as measured by pQCT. A total of 4 fracture cases were documented during follow up, including 3 subjects in the control group and 1 in the TCC group. CONCLUSIONS: This is the first prospective and randomized study to show that a programmed TCC exercise intervention is beneficial for retarding bone loss in weight-bearing bones in early postmenopausal women. Long-term follow-up is needed to substantiate the role of TCC exercise in the prevention of osteoporosis and its related fracture. PMID- 15129395 TI - Use and satisfaction with prosthetic limb devices and related services. AB - OBJECTIVES: To examine the use and satisfaction with prosthetic limb devices and satisfaction with prosthetist services in a large and diverse sample of persons with limb loss. DESIGN: Retrospective cohort study. SETTING: General community. PARTICIPANTS: Persons aged 18 to 84 years identified from the Amputee Coalition of America registry as having a major upper- or lower-limb loss due to vascular disease, trauma, or malignancy. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Use and satisfaction with prosthetic limb devices and satisfaction with prosthetists' services, assessed via structured telephone interviews. RESULTS: Most persons (94.5%) surveyed had a prosthesis and used it extensively (71h/wk). Most persons with amputations appeared to be satisfied with the overall performance of their prostheses (75.7%). Nearly one third of them, however, expressed dissatisfaction with their prostheses' comfort. Frequency of prosthesis use and satisfaction with the device were significantly higher among those with shorter timing to first prosthesis fitting, even after controlling for a wide array of respondents' sociodemographic and amputation characteristics. Overall, persons with amputations in our sample had positive assessments of their prosthetists' quality. Less favorable ratings concerned items related to the prosthetists' interpersonal skills. Multivariate analyses showed that men and black persons with amputations were less likely than their female or white counterparts to have favorable perceptions about their prosthetists across all dimensions of provider quality. Persons with fewer years of schooling were also less likely to be satisfied with their prosthetist's interpersonal manner. There were no significant differences in prosthesis use, satisfaction, or assessment of prosthetists' quality based on amputation etiology or amputation level. CONCLUSIONS: Efforts should be directed at minimizing the interval from surgery to first prosthesis fitting and at improving communication between patients and prosthetists, to improve the quality of care provided to the growing numbers of persons with limb loss. PMID- 15129396 TI - The Trinity Amputation and Prosthesis Experience Scales and quality of life in people with lower-limb amputation. AB - OBJECTIVES: To undertake preliminary research into quality of life (QOL) for a group of people with a lower-limb amputation and to investigate what aspects of the "prosthetic experience" are most strongly associated with QOL using the Trinity Amputation and Prosthesis Experience Scales (TAPES). DESIGN: Cross sectional survey. SETTING: Prosthetic limb fitting center. PARTICIPANTS: Sixty three people older than 18 years with unilateral lower-limb amputation. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The TAPES and the World Health Organization Quality of Life Questionnaire-Brief Version. RESULTS: There were no significant differences in any of the QOL domain scores (physical health, psychological, social relationships, environmental) arising from age, gender, level of amputation, or cause of amputation. However, there were significant differences depending on the length of time living with the prosthesis and the degree of prosthetic use. Stepwise regression identified different significant predictors for each domain of QOL. CONCLUSIONS: These findings support the claim that the TAPES can be used to evaluate QOL for this patient group. Further research is warranted to learn how sensitive the scale and its items are to change in clinical status. PMID- 15129397 TI - Transcranial magnetic stimulation for diagnosis of residual limb neuromas. AB - OBJECTIVE: To analyze the mechanism and examine the potential diagnostic contribution of transcranial magnetic stimulation (TMS) in diagnosing painful, clonic, and/or autonomic manifestations in amputees' residual limbs. DESIGN: Prospective study. SETTING: Regional rehabilitation institute at a medical school in France. PARTICIPANTS: Thirty-four amputees (24 with myoclonus, stump pain, or trophic skin disorders; 10 controls with no stump symptoms). INTERVENTIONS: Not applicable. MAIN OUTCOMES MEASURES: TMS performed before and after correcting prosthesis adaptation. RESULTS: TMS induced pain in 12 amputees, clonic manifestations in 4, and autonomic manifestations in 2. Twelve patients underwent magnetic resonance imaging that showed neuromas in all 12. After neuroma resection in 9 amputees, TMS no longer provoked abnormal manifestations. TMS did provoke abnormal manifestations after resection in 3 patients who had postoperative recurrent neuromas. The response to TMS was negative in subjects with a clinically silent neuroma (n=7). The response was also negative in all patients with other stump anomalies whose clinical manifestations fully regressed after conservative treatment. There was a highly significant correlation between the presence of a pathologic neuroma and TMS-induced abnormal manifestations (P<.0001). CONCLUSIONS: TMS can provoke symptoms in patients who experience spontaneous or evoked symptoms related to a neuroma. Induced symptoms are proportional to spontaneous symptoms. Removing the neuroma can stop stump symptoms and reverse the TMS effect. The response to TMS was negative in the control subjects with clinically silent neuromas; conservative treatment was successful in these cases. TMS-induced abnormal manifestations underlying mechanisms are discussed (ephaptic transmission in neuromas). PMID- 15129398 TI - Reliability, validity, and responsiveness of the locomotor capabilities index in adults with lower-limb amputation undergoing prosthetic training. AB - OBJECTIVE: To assess the reliability, validity, and responsiveness of both the standard and revised Locomotor Capabilities Index (LCI) in people with lower-limb amputation who undergo prosthetic training. DESIGN: Reliability and validity study. SETTING: Two freestanding rehabilitation centers. PARTICIPANTS: Fifty inpatients with a recent unilateral lower-limb amputation. INTERVENTIONS: Not applicable. Main outcome measures The standard LCI and a new version with a 5 level ordinal scale (LCI-5) were tested for internal consistency, test-retest reliability, ceiling effect, and effect size. The construct validity of both versions was analyzed by correlation with the Rivermead Mobility Index, a timed walking test, and the FIM instrument. RESULTS: The Cronbach alpha of both LCI versions was.95. The item-to-total correlations (Spearman rho) ranged from.50 to.87 (P<.0001 for all). The percent agreement and kappa values for the item scores ranged, respectively, from 78.4% to 100% and.58 to 1.00 in the LCI, and from 75.7% to 97.3% and.54 to.96 in the LCI-5. The intraclass correlation coefficient (model 2,1) for the total scores was.98 for both versions; the Bland Altman plot revealed no systematic trend for either version. Both the LCI and LCI 5 correlated with all criterion measures (rho range,.61-.76), with the LCI-5 showing a larger effect size during the rehabilitation period and a lower ceiling effect. Patients with transtibial amputation were more independent in performing activities than were those with transfemoral amputation; their locomotor capability negatively correlated with age. CONCLUSIONS: Both the LCI and LCI-5 captured the global locomotor ability of people with lower-limb amputation during prosthetic training. The new LCI-5 presents similar and sometimes better psychometric properties than the standard LCI. PMID- 15129399 TI - Relationships between level of disability and receipt of preventive health services. AB - OBJECTIVES: To examine relationships between level of disability and receipt of certain preventive health services, including demographic and systems variables that may explain or confound these relationships. DESIGN: Analysis of recent (1998 and 2000) data from the Behavioral Risk Factor Surveillance System, a nationwide telephone survey. SETTING: States reporting data on disability (13 in 1998, 18 in 2000). PARTICIPANTS: Noninstitutionalized persons living in the community. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Frequency of receipt of recommended preventive health services, including colorectal, cervical, and breast cancer screening, and influenza and pneumococcal vaccination. An ordinal index of disability severity was constructed from questions on activity limitations. RESULTS: In 2000, people with mild and moderate disability received influenza and pneumonia vaccinations somewhat more frequently than people without disabilities, but people with the most severe disabilities least frequently received vaccinations that year. Disabled women received fewer Papanicolaou tests and clinical breast examinations, but significant differences did not occur for mammograms in 2000. Fewer differences as a function of disability level were apparent in 2000 than 1998. Demographic variables affected receipt of most preventive services. Access to routine checkups affected all preventive services independent of disability level in both years. CONCLUSIONS: Severity of disability is related to receipt of certain preventive services but not necessarily in a simple or unidirectional way. Regardless of disability, receipt of a checkup was an important determinant of receipt of preventive health services. For almost all services and groups studied, preventive care remained below targeted goals for Healthy People 2010. PMID- 15129400 TI - Assessing pain in patients with severe cerebral palsy: development, reliability, and validity of a pain assessment instrument for cerebral palsy. AB - OBJECTIVES: To develop the Pain Assessment Instrument for Cerebral Palsy (PAICP) and to study its test-retest reproducibility and construct validity. DESIGN: Cross-sectional validation study. SETTING: Homes for severely handicapped. PARTICIPANTS: A total of 164 adults with severe cerebral palsy (CP), caregivers, and physiotherapists, and 9 healthy children. INTERVENTIONS: The PAICP contains drawings of situations, some situations of which usually produce pain. Patients rate the pain associated with each activity using a Faces Pain Scale. Reproducibility and construct validity was assessed in a pilot study with CP patients and healthy children. Construct validity and agreement between the pain scores of the patients and proxies was assessed in 160 patients with severe CP. MAIN OUTCOME MEASURE: Pain score on the PAICP. RESULTS: The measure showed adequate test-retest reproducibility. A significant difference was found between the mean scores for "painful" and "not painful" situations. We also found moderate agreement between the scores of the patients and proxies for daily activities but only for those activities in which the proxies were personally involved. CONCLUSIONS: The PAICP has adequate test-retest reproducibility and construct validity. It provides an indication of the pain experienced by patients in situations in which proxies are not personally involved and may also be more valid than proxy measures for other situations. PMID- 15129401 TI - Walking ability and its relationship to lower-extremity muscle strength in children with idiopathic inflammatory myopathies. AB - OBJECTIVE: To describe gait deficits and their association with lower-extremity muscle strength in children with juvenile idiopathic inflammatory myopathies (IIM). DESIGN: Cross-sectional, descriptive study. SETTING: Clinical research center. PARTICIPANTS: Consecutive sample of 25 ambulatory children diagnosed with juvenile IIM. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Manual muscle test (MMT) of bilateral hip flexor, extensor, and abductor; knee extensor; and ankle plantarflexor strength, all measured on a 0- to 10-point scale and summary strength measures. Video-based movement analysis to determine walking speed; gait cycle time; right and left step time; stride length; right and left step length; and stance, swing, and double-limb support phase durations. RESULTS: Walking speed (1.03+/-0.27 m/s) was reduced because of shortened stride lengths (1.03+/ 0.21 m) more than prolonged gait cycle times (1.05+/-0.22s). Walking speed highly correlated with the number of muscle groups weaker than grade 7 out of 10 (r= .89) and the strength of the hip flexors (r=.85). CONCLUSIONS: Lower-extremity strength measures, including MMT scores of individual muscle groups and the number of weak muscle groups, were predictive of gait limitations in children with juvenile IIM. PMID- 15129402 TI - Osteogenesis imperfecta in childhood: impairment and disability. A prospective study with 4-year follow-up. AB - OBJECTIVES: To study (1). changes in anthropometrics, joint range of motion (ROM), muscle strength, functional ability, caregiver assistance, and level of ambulation in children with osteogenesis imperfecta (OI) and (2). the prediction of clinical characteristics at the level of ambulation at follow-up and the prediction of clinical characteristics on progression or regression at the level of ambulation over time. DESIGN: Prospective study with follow-up of 4 years. SETTING: A children's hospital that serves a nationwide center for treatment and research in children with OI in the Netherlands. PARTICIPANTS: At follow-up, 49 children (24 boys, 25 girls; mean age +/- standard deviation, 11.3+/-3.8y; range, 5.2-19.4y) participated. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Anthropometry, joint ROM, muscle strength, fracture frequency, intramedullary rodding, level of ambulation, functional ability, and caregiver assistance. RESULTS: In type I OI, total joint ROM decreased significantly over time, especially in the lower extremities, with a significant decrease in generalized joint hypermobility according to Bulbena (median start, 7.5; interquartile range [IQR], 4-9; median end, 6; IQR, 2-7; P<.001). In types III and IV, a severe decrease in total joint ROM was present without significant changes over time. No significant changes in total muscle strength (upper or lower extremities) in the different types of OI were measured at follow-up. In OI type I, a significant increase in self-care (P=.003) and social function (P=.008) was measured; in type III, a significant increase in self-care (P=.003), mobility (P=.004), and social function (P=.005) was measured, with a significant decrease in parental assistance in self-care (P=.02) and mobility (P=.005). In type IV, a significant increase was observed in the self-care (P=.01) and social function domains (P=.02). Type of OI (regression coefficient=-1.0; 95% confidence interval [CI], 1.64 to -0.47) and total muscle strength were the only significant predictors for level of ambulation (regression coefficient=.01; 95% CI,.17-.32). Body weight was significantly lower in the group that progressed in level of ambulation (P=.03), whereas children with a decline in level of ambulation had significantly higher body weight (P=.05). CONCLUSIONS: Ours is the first study with a long-term follow up that provides information concerning the natural course of developmental outcome parameters of OI in childhood. Joint ROM and muscle strength did not change significantly over time, possibly because of the biomechanical skeletal properties of the different OI types. Functional ability improved significantly over time, but, especially in types III and IV, did not reach normative values, possibly because of a plateau phase in functional ability. Knowledge of the natural course of the disease is essential to interpret the results from intervention studies. PMID- 15129403 TI - Plasma precursors of brain catecholaminergic and serotonergic neurotransmitters in rehabilitation patients with ischemic stroke. AB - OBJECTIVE: To determine levels of plasma amino acid tyrosine and tryptophan, precursors of brain catecholamine and serotonin neurotransmitters, respectively, in rehabilitative patients with ischemic stroke. DESIGN: Controlled, pre-post analysis, consecutive sample. SETTING: Rehabilitation center. PARTICIPANTS: Twenty men with ischemic stroke (age, 68+/-11.3y) consecutively admitted into rehabilitation 15+/-10 days (range, 7-28d) after an acute cerebrovascular insult; 15 healthy sedentary subjects (controls 1); and 13 healthy hypoactive individuals who had recently had knee arthroplasty (controls 2). Both control groups were matched to stroke subjects for age, gender, and body weight. INTERVENTIONS: At 8:00 am, after overnight fasting, venous blood samples were drawn from patients to determine plasma tyrosine and tryptophan levels. A nutritional evaluation, including nitrogen balance, was made. The same procedures were repeated after 45 days of rehabilitation. Amino acid data were compared with those obtained from the controls. MAIN OUTCOME MEASURE: Plasma concentrations of amino acids. RESULTS: Patients with ischemic stroke, on admission, had lower plasma tyrosine concentration than did both controls 1 (P<.0005) and controls 2 (P<.001), but a similar tryptophan level. The plasma content of tyrosine was similar between the 2 control groups. After 45 days of rehabilitation, the stroke patients' tyrosine and tryptophan levels remained virtually unchanged, as did nutritional parameters. Nutritional intakes were adequate to meet body needs but insufficient to correct plasma tyrosine. CONCLUSIONS: Patients experiencing a recent stroke may have low plasma tyrosine levels and, therefore, reduced brain catecholamine formation. It is possible that an imbalance of brain neurotransmitters may occur. PMID- 15129404 TI - Learning to perform wheelchair wheelies: comparison of 2 training strategies. AB - OBJECTIVE: To test the hypotheses that, during wheelie training, adding the proactive balance strategy (PBS) to the conventional reactive balance strategy (RBS) increases the success rate, decreases training time, and lessens postural sway during the wheelie. DESIGN: Randomized controlled trial. SETTING: Kinesiologic laboratory in a rehabilitation center. PARTICIPANTS: Twenty-two participants (12 wheelchair users, 10 able-bodied) randomly assigned to the RBS group or to the PBS+RBS group; the groups were balanced according to gender, age, and diagnostic category. INTERVENTION: Participants were trained to perform stationary wheelies in 2 highly structured settings. MAIN OUTCOME MEASURES: Success rate in achieving wheelie competence, training time, and postural sway on a force platform. RESULTS: Success rate was 100% for both groups. The RBS group required a mean +/- standard deviation of 43+/-31 minutes of training time and the PBS+RBS group required 45+/-28 minutes (P=.54). There was no significant difference between training groups with respect to postural sway measures (P=.50). Age correlated with both training time (r=.70, P=.001) and postural sway (r=.52, P=.03). Qualitatively, we identified 3 take-off patterns and confirmed the use of the 2 balance patterns. CONCLUSIONS: The addition of the PBS to RBS training did not improve wheelie success rate, training time, or postural sway. Although older wheelchair users require more training time, many such users can learn this useful skill if given the opportunity. PMID- 15129405 TI - The wheelchair skills test (version 2.4): measurement properties. AB - OBJECTIVE: To evaluate the measurement properties of the Wheelchair Skills Test (WST), version 2.4. DESIGN: Cohort study. SETTING: Rehabilitation center. PARTICIPANTS: A total of 298 subjects (169 wheelchair users, 129 able-bodied subjects) ranging in age from 17 to 88 years. INTERVENTION: We videotaped subjects as they attempted the 50 skills of the WST 2.4. MAIN OUTCOME MEASURES: The test-retest, intrarater, and interrater reliabilities were determined on a subset of 20 wheelchair users. We assessed construct validity by evaluating whether the WST detected expected changes and concurrent validity by seeing how well total WST scores correlated with criterion measures. RESULTS: The mean time +/- standard deviation taken to administer the WST was 27.0+/-9.3 minutes. There were no serious adverse incidents, and the test was well tolerated. For the test retest, intrarater and interrater reliabilities, the intraclass correlation coefficients for the total scores were.904,.959, and.968. For individual skills, the percentage concordance ranged from 73% to 100%. Regarding construct validity, there was a slightly negative Pearson correlation between total WST score and age (-.434). Gender was identified as a significant factor on multiple regression analysis (P<.001). Wheelchair users with more than 21 days of experience scored higher than those with less experience (65.0% vs 59.6%; P=.01). Participants with stroke and related disorders had a mean score (55.0%+/-13.9%) that was significantly lower than those in other diagnostic categories (P<.05). Participants using conventional wheelchairs had lower scores than those in lightweight ones (66.4% vs 75.1%; P<.001). Regarding concurrent validity, Spearman rank correlations between total WST scores and the global assessments of the wheelchair users' therapists and admission and discharge FIM instrument scores were.394,.38, and.31. CONCLUSIONS: The WST 2.4 is practical and safe, and its measurement properties are very good to excellent. Further study is needed to determine its usefulness in various clinical settings. PMID- 15129406 TI - Durability, value, and reliability of selected electric powered wheelchairs. AB - OBJECTIVE: To compare the durability, value, and reliability of selected electric powered wheelchairs (EPWs), purchased in 1998. DESIGN: Engineering standards tests of quality and performance. SETTING: A rehabilitation engineering center. SPECIMENS: Fifteen EPWs: 3 each of the Jazzy, Quickie, Lancer, Arrow, and Chairman models. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Wheelchairs were evaluated for durability (lifespan), value (durability, cost), and reliability (rate of repairs) using 2-drum and curb-drop machines in accordance with the standards of the American National Standards Institute and Rehabilitation Engineering and Assistive Technology Society of North America. RESULTS: The 5 brands differed significantly (P75% for all three genes). Methylation of CD44 and EDNRB was observed in 41 and 38% of tumors but was not present in HGPIN. CONCLUSIONS: These data suggest that genes may be methylated at different points in the histopathologic progression of prostate cancer and these differences can be found in various histologic foci from the same gland. PMID- 15129427 TI - Immunological monitoring during combination of patient-oriented peptide vaccination and estramustine phosphate in patients with metastatic hormone refractory prostate cancer. AB - BACKGROUND: Additive antitumor effects could be achieved by combination of immunotherapy and cytotoxic agents with no or minimum suppression. METHODS: Thirteen patients positive for human leukocyte antigen (HLA)-A24 or -A2 with metastatic hormone refractory prostate cancer (HRPC) who had failed to respond to the prior-peptide vaccination were entered in the combined peptide vaccination and estramustine phosphate. Conducted immune monitoring on those 13 patients were mainly peptide-specific cytotoxic T lymphocyte (CTL) precursor analysis by IFN gamma productions and peptide-reactive IgG by an enzyme-linked immunosorbent assay (ELISA). RESULTS: Grade 3 arrhythmia or cerebral infarction was observed in two cases, and Grade 1 or 2 dermatologic reaction at the vaccination sites was observed in all 13 cases. Eleven patients who received more than one cycle of treatment were eligible for immunological and clinical evaluation. There was no significant immunosuppression in most cases when the peptide and a half dose (280 mg/day) of estramustine were administrated, whereas severe immunosuppression was observed in the first two patients who received both the peptide and a full dose (560 mg/day) estramustine. Augmentation of peptide-specific CTL precursors or peptide-specific IgG was observed in 6 of 11 or 10 of 11 cases, respectively. Ten of 11 patients showed serum prostate-specific antigen (PSA) level decrease from the baseline including 8 patients with a serum PSA level decrease of > or =50%. CONCLUSIONS: These results encouraged the further evaluation of the combination of peptide vaccination and low-dose estramustine phosphate for metastatic HRPC patients. PMID- 15129428 TI - Leveling of prostate cancer mortality in Western Europe. AB - BACKGROUND: Mortality from prostate cancer (CaP) has increased throughout Europe until the early 1990s. Trends in 24 European countries, the European Union (EU), six selected Central and Eastern European countries, and the Russian Federation have been updated to 1999. METHODS: Cancer death certification data for CaP were abstracted from the World Health Organization database. RESULTS: In the EU, the peak rate (15.7/100,000) was reached in 1993, followed by a leveling off and a decline to 14.1/100,000 in 1999. Age-standardized analysis for each subsequent age group of men aged 50 or over showed larger absolute falls in the elderly. CaP rates were lower in Central and Eastern European countries providing data, but showed a rise from 9.7/100,000 in 1980 to 11.3 in 1996, and leveled off thereafter. Rates were originally lower, but the rises larger in the Russian Federation (from 5.1/100,000 in 1980 to 8.1/100,000 in 1999). In the late 1990s, there was a threefold difference between the highest rates of 22/100,000 in Norway and those of 7.7 in Russia or 7.3 in Ukraine. Such a difference was, however, restricted to the elderly, since at age 35-64 the Russian rate (6.7/100,000) was the same as that of Norway, and only Greece and Italy had appreciably lower rates. CONCLUSION: The pattern of trends in CaP rates observed across Europe is consistent with a favorable role of improved diagnosis, but mainly of advancements of therapy (including more widespread adoption of radical prostatectomy and androgen blockage) on CaP mortality in Western Europe. PMID- 15129429 TI - Potent antitumor activity after systemic delivery of a doubly fusogenic oncolytic herpes simplex virus against metastatic prostate cancer. AB - BACKGROUND: Although conventional radiation therapy and surgery are potentially curative treatments for organ-confined prostate cancer, there are few effective treatments for metastatic disease. Oncolytic viruses have shown considerable promise for the treatment of solid tumors including prostate cancer. We recently demonstrated that incorporation of a cell membrane fusion capability into an oncolytic herpes simplex virus (HSV) can significantly increase the antitumor potency of the virus. METHODS: We used a mouse model of primary and metastatic human prostate cancer established from PC-3M-Pro4 to evaluate three different types of oncolytic HSVs: non-fusogenic Baco-1, singly fusogenic Synco-2, and doubly fusogenic Synco-2D. RESULTS: Our results show that Synco-2D has greater oncolytic activity than either Baco-1 or Synco-2 virus. Against lung metastases of human prostate cancer xenografts, intravenous administration of Synco-2D had produced a significant reduction of tumor nodules by day 40 post-inoculation as compared with Synco-2 (P < 0.05), Baco-1 (P < 0.01), and PBS control (P < 0.01). CONCLUSIONS: We conclude that the doubly fusogenic Synco-2D is an effective therapeutic agent for human metastatic prostate cancer. PMID- 15129430 TI - Regulation of interleukin-6-mediated PI3K activation and neuroendocrine differentiation by androgen signaling in prostate cancer LNCaP cells. AB - BACKGROUND: Neuroendocrine (NE) differentiation in prostate cancer has been suggested to be one of the early events in the development of androgen independence. In the human prostate cancer LNCaP cell line, treatment with interleukin-6 (IL-6) induces NE-like differentiation, which is similar to the phenomena observed in advanced stages of prostate cancer progression. In this study, we investigate how androgen plays a role in IL-6-mediated NE differentiation in LNCaP cell line. METHODS: Western blot, co-immunoprecipitation (co-IP), and GST pull-down assays were performed to detect the protein expression and protein-protein interaction. PI3K kinase assay was used to measure PI3K activity. RESULTS: Addition of androgen blocks IL-6-mediated PI3K activation and NE differentiation in LNCaP cells. In vivo and in vitro protein interaction assays suggested that androgen receptor (AR) can directly interact with IL-6 transducer gp130. In addition, androgen treatment enhances the interaction between AR and gp130, interrupts the IL-6-induced gp130-mediated PI3K activation, which may lead to inhibition of IL-6-mediated NE differentiation in LNCaP cells. CONCLUSIONS: Our results suggest androgen and AR can regulate IL-6-mediated LNCaP cell NE differentiation via directly modulating the IL-6-PI3K pathway. PMID- 15129431 TI - The ErbB signaling network is coordinately expressed and activated in the mouse prostate. AB - BACKGROUND: The ErbB family of receptor tyrosine kinases contribute to human prostate cancer, however, ErbB activity in the normal prostate requires further investigation. The mouse prostate may serve as an important model system to study the molecular mechanisms regulating ErbB signaling in the prostate. METHODS: We employed RT-PCR and immunohistochemistry to perform a comprehensive expression profile of ErbB receptors and ligands in the mouse prostate. Physiological receptor activation in situ was measured by receptor phospho-tyrosine analysis. RESULTS: Expression of all four ErbB receptors and the EGF-like ligands EGF, TGF alpha, AR, beta C, Hb-EGF, NRG1, and NRG3 was detected in the mouse prostate. We failed to detect expression of the ErbB ligand, ER. Physiological receptor activation was observed within the mature mouse prostate at 10 weeks but not in the prostates of 3- or 6-week-old mice. CONCLUSIONS: Coordinated ErbB receptor and ligand expression coupled with receptor activation profiles provide strong evidence that ErbB signaling contributes to mouse prostate function. PMID- 15129432 TI - Bioreactor cultivation enhances the efficiency of human embryoid body (hEB) formation and differentiation. AB - The promise of human embryonic stem cells (hESCs) to provide an unlimited supply of cells for cell therapy and tissue engineering depends on the availability of a controllable bioprocess for their expansion and differentiation. We describe for the first time the formation of differentiating human embryoid bodies (hEBs) in rotating bioreactors to try and control their agglomeration. The efficacy of the dynamic process compared to static cultivation in Petri dishes was analyzed with respect to the yield of hEB formation and differentiation. Quantitative analyses of hEBs, DNA and protein contents, and viable cell concentration, as measures for culture cellularity and scale-up, revealed 3-fold enhancement in generation of hEBs compared to the static culture. Other metabolic indices such as glucose consumption, lactic acid production, and pH pointed to efficient cell expansion and differentiation in the dynamic cultures. The type of rotating vessel had a significant impact on the process of hEB formation and agglomeration. In the slow turning lateral vessel (STLV), hEBs were smaller in size and no large necrotic centers were seen, even after 1-month cultivation. In the high aspect rotating vessel (HARV), hEB agglomeration was massive. The appearance of representative tissues derived from the three germ layers as well as primitive neuronal tube organization, blood vessel formation, and specific-endocrine secretion indicated that the initial developmental events are not altered in the dynamically formed hEBs. Collectively, our study defines the culture conditions in which control over the aggregation of differentiating hESCs is obtained, thus enabling scaleable cell production for clinical and industrial applications. PMID- 15129433 TI - Inhibition of the Trichoderma reesei cellulases by cellobiose is strongly dependent on the nature of the substrate. AB - The inhibition effect of cellobiose on the initial stage of hydrolysis when cellobiohydrolase Cel 7A and endoglucanases Cel 7B, Cel 5A, and Cel 12A from Trichoderma reesei were acting on bacterial cellulose and amorphous cellulose that were [(3)H]- labeled at the reducing end was quantified. The apparent competitive inhibition constant (K(i)) for Cel 7A on [(3)H]-bacterial cellulose was found to be 1.6 +/- 0.5 mM, 100-fold higher than that for Cel 7A acting on low-molecular-weight model substrates. The hydrolysis of [(3)H]-amorphous cellulose by endoglucanases was even less affected by cellobiose inhibition with apparent K(i) values of 11 +/- 3 mM and 34 +/- 6 mM for Cel 7B and Cel 5A, respectively. Contrary to the case for the other enzymes studied, the release of radioactive label by Cel 12A was stimulated by cellobiose, possibly due to a more pronounced transglycosylating activity. Theoretical analysis of the inhibition of Cel 7A by cellobiose predicted an inhibition analogous to that of mixed type with two limiting cases, competitive inhibition if the prevalent enzyme-substrate complex without inhibitor is productive and conventional mixed type when the prevalent enzyme-substrate complex is nonproductive. PMID- 15129434 TI - Partitioning of model toxins to hydrophobically terminated DAB dendrimers. AB - Dendrimers are attractive in biological and biomedical applications due to the similarity in their molecular size to biologically relevant molecules and the large number of chain ends available functionalization. In the current work, we examined the potential of diamino butane (DAB) dendrimers functionalized with long alkyl chains as partitioning agents for hydrophobic toxins for use as a prefiltering stage in a bioartiticial liver. DAB dendrimers of various generations that had been previously fully modified with palmitoyl chloride were obtained. A study of the kinetics of partitioning of acetylsalicylic acid (ASA) suggested that while significant toxin removal occurred in 30 s, although a slight time dependent increase in removal was noted up to 60 minutes. The partitioning of 6 hydrophobic toxins from aqueous solution to the modified dendrimers in 30 minutes was examined. The results demonstrated that a number of factors, including the pKa of the toxin, its octanol water partitioning coefficient and molecular size contributed to the level of toxin removal. Toxin removal on a molar basis increased with increasing dendrimer generation for all toxins, with the modified G5 dendrimers partitioning 50-100 toxin molecules in most cases. Dendrimer modification with C4 alkyl chains rather than Cl5 chains significantly decreased toxin removal, although chains longer than C10 seemed to partition equal amounts of toxins. The results of the study demonstrate that water-soluble dendrimers modified with hydrophobic end groups may be useful for the removal of toxins from the blood in a prereaction step for a bioartificial liver, but that a better understanding of the molecular mechanisms of removal may be necessary before it is possible to predict the levels of toxin removal. PMID- 15129436 TI - Modeling and simulation of oxygen-limited partial nitritation in a membrane assisted bioreactor (MBR). AB - Combination of a partial nitritation process and an anaerobic ammonium oxidation process for the treatment of sludge reject water has some general cost-efficient advantages compared to nitrification-denitrification. The integrated process features two-stage autotrophic conversion of ammonium via nitrite to dinitrogen gas with lower demand for oxygen and no external carbon requirement. A nitrifying membrane-assisted bioreactor (MBR) for the treatment of sludge reject water was operated under continuous aeration at low dissolved oxygen (DO) concentrations with the purpose of generating nitrite accumulation. Microfiltration was applied to allow a high sludge retention time (SRT), resulting in a stable partial nitritation process. During start-up of the MBR, oxygen-limited conditions were induced by increasing the ammonium loading rate and decreasing the oxygen transfer. At a loading rate of 0.9 kg N m(-3) d(-1) and an oxygen concentration below 0.1 mg DO L(-1), conversion to nitrite was close to 50% of the incoming ammonium, thereby yielding an optimal effluent within the stoichiometric requirements for subsequent anaerobic ammonium oxidation. A mathematical model for ammonium oxidation to nitrite and nitrite oxidation to nitrate was developed to describe the oxygen-limited partial nitritation process within the MBR. The model was calibrated with in situ determinations of kinetic parameters for microbial growth, reflecting the intrinsic characteristics of the ammonium oxidizing growth system at limited oxygen availability and high sludge age. The oxygen transfer coefficient (K(L)a) and the ammonium-loading rate were shown to be the appropriate operational variables to describe the experimental data accurately. The validated model was used for further steady state simulation under different operational conditions of hydraulic retention time (HRT), K(L)a, temperature and SRT, with the intention to support optimized process design. Simulation results indicated that stable nitrite production from sludge reject water was feasible with this process even at a relatively low temperature of 20 degrees C with HRT down to 0.25 days. PMID- 15129435 TI - Physiological significance of the cytometric distribution of fluorescent yeasts after viability staining. AB - This article describes a new method for the early detection of alcoholic fermentation arrest. This methodology is based on the flow cytometric assessment of Saccharomyces cerevisiae yeasts stained with a carboxyfluorescein diacetate fluorescent viability probe. Multicomponent analysis of viable cell distribution constitutes a promising new tool to describe physiological and dynamic changes to heterogeneous viable populations during alcoholic fermentation, through its ability to discriminate between successful processes and those ending prematurely. This framework, which is based on the comparison of cytometric histogram descriptors' combinations that can be related to simple physiological significance comparison, quickly and simply, allows testing yeasts for their fermentation ability and can be used to detect any kind of viability loss so that fermentation arrest can be avoided. PMID- 15129437 TI - Influence of beta-subunit on thermal and high-pressure process stability of tomato polygalacturonase. AB - Polygalacturonase (PG; E.C. 3.2.1.15) was extracted from tomato fruit and purified by cation-exchange chromatography. Two peaks containing PG activity were detected: the first denotes a thermolabile PG fraction (PG2) and the second a thermostable fraction (PG1). PG1 is a dimer of PG2 and a heat-stable protein called the beta-subunit. In contrast to its resistance to heat, PG is easily inactivated at elevated pressure. Although the thermal stability of purified tomato PG1 and PG2 is distinctly different, they show an identical pressure stability. To gain further insight into the thermal and pressure stability of both PG isoenzymes, the in vitro recombination of PG2 and beta-subunit was studied. After severe heat (up to 140 degrees C for 5 min) and pressure (up to 800 MPa for 15 min) treatments, the residual fractions containing the beta subunit were able to convert PG2 into the heat-stable PG1, showing the extreme thermal and pressure stability of the beta-subunit. PG1 was detected in heat treated tomato juice and, to a lesser extent, in tomato pieces. In contrast, as was the case for purified PG, no pressure-stable fraction was observed when tomato juice and pieces were treated under pressure. These data clearly show the differing behavior of the PG1-PG2-beta-subunit system under thermal and high pressure treatments and offer the possibility of inactivating tomato PG using high pressure without the need for high temperatures. PMID- 15129438 TI - Role of oxidative enzymatic treatments on enzymatic hydrolysis of softwood. AB - The impact of oxidative modification and partial removal of lignin by laccase mediator treatments on the enzymatic hydrolysis of steam-pretreated softwood (SPS) was evaluated. Two mediators, N-hydroxy-N-phenylacetamide (NHA) and its acetylated precursor, were oxidized by the laccase from Trametes hirsuta, and their effects on the activity of cellulolytic enzymes and on the hydrolysis yield of SPS were examined. Both simultaneous and sequential combinations of laccase mediator treatments with commercial cellulases increased the sugar yield in the enzymatic hydrolysis of SPS. The maximal increase was 21% when a sequential treatment was applied. Laccase treatment alone was also shown to improve hydrolysis. NHA oxidized by laccase inhibited significantly the cellulases of Trichoderma reesei, but the presence of the solid substrate protected the activities against oxidative inactivation. Surface analysis of the lignocellulosic substrate before and after the laccase and cellulase treatments revealed an enrichment of lignin and an increase of carboxylic groups on the surface of the hydrolysis residue. PMID- 15129439 TI - Encapsulation of crosslinked penicillin G acylase aggregates in lentikats: evaluation of a novel biocatalyst in organic media. AB - The encapsulation of crosslinked enzyme aggregates (CLEA) of penicillin G acylase into a very rigid polymeric matrix based on polyvinyl alcohol (LentiKats) has been used successfully to improve the inadequate mechanical properties of CLEA. This encapsulation decreased CLEA activity by only around 40%. As compensation, a significant improvement in the stability of the CLEA in the presence of organic solvents was detected. This could be related to the highly hydrophilic environment inside the LentiKats biocatalysts: Partition experiments showed that the concentration of dioxane inside LentiKats was lower than in the reaction medium. In fact, thermal stability was about the same as in the corresponding CLEA. This permitted great improvement in the reaction rate for thermodynamically controlled synthesis of a model antibiotic (using phenylacetic acid and 7-amino deacetoxycefalosporanic acid). Even more importantly, yields could be improved by using LentiKats-encapsulated CLEA, very likely by a favorable product/substrate partition. Thus, this very simple technique not only provides an efficient technique for solving the mechanical stability problem associated with CLEA, but also greatly improves the behavior of CLEA in organic media. PMID- 15129440 TI - Production and characterization of liquid-core capsules made from cross-linked acrylamide copolymers for biotechnological applications. AB - A novel chemistry has been developed for the production of capsules composed of a hydrophobic liquid core surrounded by a cross-linked polyacrylamide/alginate membrane. These liquid-core capsules may be used in capsular perstraction for the removal of inhibitory products from bioprocesses and bioconversions. They have the advantage of having a high surface area to promote rapid mass transfer, while separation of the organic core phase from the aqueous environment by the capsule membrane prevents the formation of stable emulsions and potential problems associated with toxicity of the organic phase for microbial cells or enzymes. Monodisperse spherical liquid-core capsules of between 800 microm and 1.6 mm diameter, with high mechanical resistance, have been prepared by co-extrusion, using the jet break-up technique. Capsules produced from a solution of MBA/total monomer (5%) were found to be more elastic and have a higher burst force when exposed to chelating agents such as phosphate or citrate. The mechanical resistance was unaffected by buffer solutions in the pH range 4-9 and after sterilization at 121 degrees C for 20 min. Capsules having membranes composed of a copolymer of acrylamide and N-hydroxymethylacrylamide exhibited even higher mechanical stability toward chelating agents. PMID- 15129441 TI - A mixed mechanistic-electrostatic model to explain pH dependence of glycosyl hydrolase enzyme activity. AB - Glycosyl hydrolases are a vast group of enzymes that share a common topology at their active site with two acidic residues that are responsible for activity. In spite of their similarity, they exhibit a wide range of pH optima that must depend on other factors. Using structural and mechanistic knowledge about glycosyl hydrolases from families 7, 10, and 16, we have formulated a new mathematical model that can include not only the ionization behavior of the catalytic residues but also as many ionizable residues as desired in the active site. In addition, the model can incorporate electrostatic influences via acid dissociation equilibrium constants and chemical relationships such as hydrogen bonds. The results of the simulations indicate a clear shift in the pH dependence of activity for the enzymes only when a close interrelation (hydrogen bond) between the catalytic and auxiliary residues in the active site is taken into account. This explains the observations from mutagenesis studies that show this type of shift and cannot be explained by a purely electrostatic interaction theory. Moreover, the presence of the kind of chemical interaction proposed could provide stabilization of the activity in the presence of environmental, structural, pH and electrostatic variations. These findings and the implications for the design of new mutagenesis strategies are discussed. The results suggest a way to modify, via site-directed mutagenesis, the acid dissociation of one of the catalytic residues in the active site independently of the other, which could have clear advantages over the purely electrostatic modifications that usually affect both residues simultaneously. PMID- 15129442 TI - Fermentation of biomass-generated producer gas to ethanol. AB - The development of low-cost, sustainable, and renewable energy sources has been a major focus since the 1970s. Fuel-grade ethanol is one energy source that has great potential for being generated from biomass. The demonstration of the fermentation of biomass-generated producer gas to ethanol is the major focus of this article in addition to assessing the effects of producer gas on the fermentation process. In this work, producer gas (primarily CO, CO(2), CH(4), H(2), and N(2)) was generated from switchgrass via gasification. The fluidized bed gasifier generated gas with a composition of 56.8% N(2), 14.7% CO, 16.5% CO(2), 4.4% H(2), and 4.2% CH(4). The producer gas was utilized in a 4-L bioreactor to generate ethanol and other products via fermentation using a novel clostridial bacterium. The effects of biomass-generated producer gas on cell concentration, hydrogen uptake, and acid/alcohol production are shown in comparison with "clean" bottled gases of similar compositions for CO, CO(2), and H(2). The successful implementation of generating producer gas from biomass and then fermenting the producer gas to ethanol was demonstrated. Several key findings following the introduction of producer gas included: (1) the cells stopped growing but were still viable, (2) ethanol was primarily produced once the cells stopped growing (ethanol is nongrowth associated), (3) H(2) utilization stopped, and (4) cells began growing again if "clean" bottled gases were introduced following exposure to the producer gas. PMID- 15129443 TI - Novel synthetic jasmonates as highly efficient elicitors for taxoid production by suspension cultures of Taxus chinensis. AB - Suspension cultures of Taxus chinensis were used as a model plant cell system to evaluate novel synthetic jasmonates as elicitors for stimulating the biosynthesis of secondary metabolites. Significant increases in accumulation of taxuyunnanine C (Tc) were observed in the presence of newly synthesized 2-hydroxyethyl jasmonate (HEJA) and trifluoroethyl jasmonate (TFEJA) without their inhibition on cell growth. Addition of 100 microM HEJA or TFEJA on day 7 led to a high Tc content of 44.3 +/- 1.1mg/g or 39.7 +/- 1.1 mg/g (at day 21), while the Tc content was 14.0 +/- 0.1 mg/g and 32.4 +/- 1.6 mg/g for the control and that with addition of 100 microM methyl jasmonate (MJA), respectively. The superior stimulating ability of HEJA and TFEJA over MJA, which was generally considered as the best chemical for eliciting taxoid biosynthesis, suggests that the novel jasmonate analogues may have great potential in application to other cell culture systems for effcient elicitation of plant secondary metabolites. PMID- 15129445 TI - [Lumbosacral angiolipoma]. PMID- 15129446 TI - When surgeons become SARS patients. PMID- 15129447 TI - Government cracks down on billing fraud. AB - As the largest professional caregiving group, nurses have a stake in preventing misuse of the reimbursement system. Technology also plays a crucial role, as automated billing systems can both prevent and enable billing fraud. PMID- 15129448 TI - Quick-and-easy bed status update. AB - Increase communication with department heads and administration regarding bed availability and house census. PMID- 15129449 TI - Restraint, seclusion standards raise questions. AB - Several of the Joint Commission's new restraint and seclusion standards have prompted caregivers to seek additional clarification. PMID- 15129450 TI - Competent i.v. management, Part 1. AB - Review two court cases regarding common pitfalls and current legal trends of i.v. management. PMID- 15129451 TI - Salary survey 2001. PMID- 15129452 TI - Staying power. AB - First-line managers influence nurses' job satisfaction and decrease nurses' interest in union activity. PMID- 15129453 TI - Managing burn emergencies. AB - Learn how to provide emergency care for a burn patient, from safety at the scene to patient stabilization, and, if necessary, transport to a burn center. PMID- 15129454 TI - Increase input, optimize output. PMID- 15129455 TI - High-tech, high-touch care. AB - Follow the expansion and transformation of Good Samaritan Hospital's 46-bed NICU to a 21,250-square-foot newly designed level III unit. PMID- 15129456 TI - Pro-patient partnerships. AB - The degree to which nurses and physicians collaborate greatly impacts patient care. These elements of effective partnerships and strategies can help you achieve mutual benefits. PMID- 15129457 TI - Automating the medication system: yesterday's impossibilities are today's realities. AB - Automating previously manual functions helps reduce the potential for medical errors by introducing objective standards and procedures into the system. PMID- 15129458 TI - Comorbidities in general practice. PMID- 15129459 TI - Summer skin check. PMID- 15129460 TI - CBS and CBT. PMID- 15129461 TI - Managing 'metabolic syndrome' and multiple risk factors. AB - BACKGROUND: Risk factors tend to cluster and are shared across common diseases seen in general practice. The 'metabolic syndrome' is a cluster of fasting hyperglycaemia, abdominal adiposity, dyslipidaemia and hypertension. This syndrome is associated with both insulin resistance and behaviourally modifiable risk factors such as smoking, physical activity and unhealthy diet. OBJECTIVE: This article aims to provide pragmatic guidance on conditions that are lifestyle based and present as a number of disease states that require multiple interventions. Management of comorbidity and multiple risk factors is discussed using a case vignette. DISCUSSION: Metabolic disease states have common bases and their management is directed toward identifying all the risk factors, establishing absolute risk and intervening sequentially. PMID- 15129462 TI - A smoking related triad: PAD, COPD and CCF. AB - BACKGROUND: Tobacco smoking is the modifiable risk factor responsible for the greatest burden of disease (loss of health and premature mortality) in Australia. OBJECTIVE: This article discusses the complexities of management of a patient suffering from a smoking related triad of illnesses: peripheral arterial disease, chronic obstructive pulmonary disease and ischaemic heart disease related congestive cardiac failure. DISCUSSION: As well as considering optimal management of the individual conditions, the patient's mental health and overall quality of life needs to be considered. Even at this late stage, smoking cessation is critically important. The patient is likely to require 6-8 medications, creating the potential for interactions and confusion, so careful medication management and patient education is required. The support of a multidisciplinary team will be required, with coordination of care being the key. Care planning and case conferencing between the general practitioner, the patient and other health professionals is essential for optimal care. PMID- 15129463 TI - Falls, osteoporosis and atrial fibrillation. AB - BACKGROUND: Poly-morbidity is the hallmark of the frail elderly. The traditional model of treating each disease on its own merit does not work for this patient group. OBJECTIVE: This article outlines the assessment, investigations and management of falls and atrial fibrillation in the elderly. DISCUSSION: The challenge for the general practitioner lies in weighing up the potential benefits and harms of the 'curative' approach against those of the 'palliative' approach. The key concern should always be the patient's desired quality of life. Forgoing largely unproven 'medical heroics' in favour of supportive services in association with available community resources helps to maintain the patient's independence and dignity. PMID- 15129464 TI - Psychiatric comorbidity in general practice. AB - BACKGROUND: Much of teaching and research has concentrated on single disease entities. In general practice, however, many patients suffer from a number of interacting illnesses at the same time. It is unlikely we will ever have randomised controlled trials to guide the management of such patients. OBJECTIVE: This article aims to develop a framework that will assist general practitioners in day-to-day clinical work with patients presenting with concurrent multiple physical and mental health problems. DISCUSSION: A positive, caring, patient centred approach is required and multiple diagnoses need to be managed within an integrated treatment plan. The quality of the therapeutic relationship is central to patient outcomes. Assessing and managing risk issues is a priority and coordinating care with other health professionals is essential. Set realistic goals, don't make changes too quickly or all at once, and don't assume that because a patient is taking a particular medication the associated diagnosis is correct. PMID- 15129465 TI - Managing schizophrenia in general practice. AB - BACKGROUND: The lifetime risk of schizophrenia is about one in 100. It is an illness associated with substantial disability and increased risk of physical illness. OBJECTIVE: This article updates general practitioners on the physical, psychological and social management of patients with schizophrenia. DISCUSSION: General practitioners have a key role in managing patients with schizophrenia, including early detection of prodromal symptoms, monitoring and preventing relapses, and providing high quality primary and secondary prevention for common physical problems. Effective care requires a thorough understanding of local specialist services, pharmacotherapies, and familiarity with psychosocial interventions that will genuinely benefit patients. PMID- 15129466 TI - Cannabis and psychosis. AB - BACKGROUND: Cannabis use is commonly identified in people who present with psychosis. OBJECTIVE: This case study aims to provide a practical approach for general practitioners seeing patients with comorbid cannabis and mental health concerns. DISCUSSION: Cannabis related comorbidity is commonly seen in general practice. General practitioners can manage most presentations and help to reduce the likely occurrence of cannabis induced psychosis through the use of psychosocial support, brief interventions and harm minimisation. PMID- 15129467 TI - Patient education. Gait re-training exercises. PMID- 15129469 TI - Hyperinsulinaemia. PMID- 15129468 TI - Consensus statement for the prevention of vascular disease. AB - This consensus statement for the prevention of vascular disease in people over 50 years of age aims to consolidate key messages from a number of evidence based guidelines and studies. It addresses the assessment and principles of management of risk factors for vascular disease, including those developed by Diabetes Australia, Kidney Health Australia, the National Heart Foundation of Australia, and the National Stroke Foundation of Australia. For more detailed information, particularly concerning treatments and levels of evidence to support the recommendations outlined in this statement, refer to the source guidelines and literature (see References). PMID- 15129471 TI - Gastrointestinal health. The role of pro- and pre-biotics in standard foods. AB - BACKGROUND: Diarrhoea is a common symptom of gastrointestinal illness, ranking among the 20 most frequent reasons for patient encounters in general practice in Australia. Children under 5 years of age are especially vulnerable to bacterial gastroenteritis such as infections with Campylobacter jejuni. Everyday diet, in particular the regular consumption of pro- and pre-biotics, can help with the maintenance of a healthy gastrointestinal system. OBJECTIVE: This article outlines the potential of 'everyday standard' food items such as cheese to promote healthy gastrointestinal microflora and to prevent gastrointestinal illness such as diarrhoea. DISCUSSION: Some common food items such as yoghurt, sauerkraut, garlic and cheese contain probiotics in the form of live lactic acid bacteria, and/or prebiotics in the form of fructans, a dietary fibre. Cheese contains both probiotic bacteria and the prebiotic dietary fibre inulin. The regular consumption of cheese has been associated with a reduction in the risk of Campylobacter enteritis. PMID- 15129470 TI - The funnel web and common spider bites. AB - BACKGROUND: The funnel web spiders, encompassed within the genera Atrax and Hadronyche, are the most dangerous spiders in the world. Although the incidence of envenomation is low, funnel web spiders remain a cause of considerable public concern. However, most common spider bites produce only minor effects-requiring only symptomatic treatment. OBJECTIVE: This article describes the clinical features and treatment of the funnel web spider and that of its close relative, the mouse spiders. It also covers the question of necrotising arachnidism as well as bites from other common species of spiders. DISCUSSION: Appropriate first aid combined with the administration of specific antivenom can be life saving for funnel web spider bites. True necrotising arachnidism appears to be rare. If suspected, clinicians must first consider the very wide differential diagnoses. PMID- 15129472 TI - Obesity case study. Eye series--14. PMID- 15129473 TI - Risk management. Discussing benefits and risks with patients. PSA testing. AB - Obtaining valid consent from patients is good medical practice and a legal necessity. The process of obtaining consent involves shared decision making and a discussion of benefits and risks with patients. This article uses the example of PSA screening to explore the issue of shared decision making and outlines some strategies for general practitioners to use when discussing risks with patients. PMID- 15129474 TI - The collaborative method. A strategy for improving Australian general practice. AB - BACKGROUND: The United Kingdom National Primary Care Collaborative (NPCC) is said to be the largest health service improvement project in the world. The 2000 practices participating serve 11.5 million patients throughout the UK. Improvements such as a 60% reduction in waiting times to see a general practitioner and a four-fold reduction in coronary heart disease mortality have drawn the attention of policy makers in Australia. In 2003, I accompanied a group of Australians to Manchester in the UK to participate in an international training course in collaboratives hosted by the NPCC. The Australian government has recently called for tenders to run the Australian Primary Care Collaborative (APCC) program. OBJECTIVE: To describe the NPCC and discuss how the methodology could be adapted and applied to Australia. DISCUSSION: The Australian primary health care system faces challenges in application of evidence to patient care. Collaborative methodology warrants consideration to address these. Differences including distance, business issues, remuneration mechanisms, infrastructure deficiencies and clinician cynicism present challenges in adaptation and application to Australia. Given adequate support, an APCC could empower general practices to continue to make improvements in many areas of activity. PMID- 15129475 TI - Simulation training in emergency medicine. An important need for primary care training. PMID- 15129476 TI - Patients' views on the training of medical students in Australian general practice settings. AB - AIM: To explore patients' views on training medical students in their general practice. METHOD AND SETTING: Consenting patients attending eight urban teaching practices completing a self administered survey before and after the consultation. RESULTS: One hundred and four patients attended for appointments: 94 consented to the involvement of a medical student, 88 completed surveys before and after their consultation (response rate 85%), 80% said the main reason for consenting was to benefit the student, and 70% said they would never refuse the presence of a medical student. Student involvement was less than that consented to: only 18 (20%) patients reported that the student independently conducted any of the consultation; 52 (59%) would accept this level of involvement in the future. DISCUSSION: Patients are a willing, but potentially under used resource for training medical students in general practice. Improved collaboration with patients would provide better teaching opportunities for students at all levels. PMID- 15129477 TI - Peer reviewed publication rates. An indication of research output. AB - BACKGROUND: The General Practice Evaluation Program (GPEP) funded general practice research between 1990-1999. We were interested in the publication rate of GPEP funded research as a measure of its research productivity. METHODS: A literature search and an email survey of GPEP researchers. We compared publication rates between the types of grants, types of institutions, and academic status of the authors. RESULTS: By June 2002, there were 201 peer reviewed articles in a range of 64 Australian and international peer reviewed journals from 99 projects (41% of completed or in progress projects, mean 2.3 per project), ranging from 0-22 per project. Forty-one investigators indicated they were in the process of writing for publication or plan to publish. They were more likely to publish with the support of a university. DISCUSSION: GPEP has achieved one of its major objectives--to contribute to evidence and knowledge about general practice. The publication rate indicates that Australian general practice research should still improve. PMID- 15129478 TI - The use of diabetes registers in an urban division of general practice. PMID- 15129479 TI - [Polymorphism of the locus DYS438 in the Polish population]. AB - This paper describes polymorphism of a new locus DYS438 in a Polish population sample of 249 males. In an analyzed population sample 5 of 9 alleles, to date identified, were found (6-14). The most frequent allele was 11 with a frequency of 0.5502. In our population bimodal frequency distribution of DYS438 alleles contrary to the other European populations was not observed. The calculated gene diversity value for DYS438 locus is 0.53 and is higher than observed for DYS391, DYS3891, DYS392 i DYS393 and lower than observed for DYS385I/II, DYS19, DYS389II, DYS439 i DYS390 Y-STR loci analyzed to date in our population. Moreover comparison of homogeneity distribution of DYS438 alleles between different populations was analyzed. Very significant statistical differences (p < 0.0000) were observed between our population and three other European population samples (Italy, Portugal, Spain and Denmark). PMID- 15129480 TI - [Effect of bovine serum albumin and restriction enzyme on quality of DNA profile of tissues extracted from paraffin blocks and histological slides]. AB - This paper reports the possibility of DNA profiling obtained from fixed and paraffin-embedded tissues or histological slides. The influence of using bovine serum albumin and restriction enzyme Hinfl on quality of DNA was analysed. The usability of those chemicals was estimated by examination of amplification results locus VWA, and multiplex PCR with the use commercial kit AmpFlSTR Identifiler (including 15 loci + amelogenine locus) and AmpFlSTR SEfiler kit (including 11 loci + amelogenine locus) Applied Biosystems. PCR products of VWA locus were separated by electrophoresis on polyacrylamide gels and visualized by silver staining. The products of loci of AmpFlSTR Identifiler and AmpFlSTR SEfiler were analyzed by capillary electrophoresis on an ABI Prism 310 sequencer. Results achieved with and without using bovine serum albumin and restriction enzyme Hinfl were compared and positive influence of those substances on DNA quality was demonstrated. Obtained results of purification DNA with the use of restriction enzyme were verified in paternity testing case. PMID- 15129481 TI - [Identification of a very short YCAII allele in the northern Polish population]. AB - The YCAII system is one of the Y-chromosomal STR markers with dinucleotide repetitive units and due to the duplication on the chromosome, its amplification provides two polymorphic products. Due to the high polymorphism, YCAII was included in the Y-STR Haplotype Reference Database (http://www.ystr.org) and significantly increased discrimination power of the collected haplotypes. During studies on the Y-STR polymorphism in the Polish population, a very short YCAII allele with relatively low stutter was identified. Sequencing revealed the presence of 11 dinucleotide repetitive units characteristic of this system. Among 3998 extended haplotypes collected in the database, this allele was found only in the population of south-western Germany. Similarities between the haplotypes possessing the YCAII*11 allele suggest their common origin. PMID- 15129482 TI - [Allele distribution of the ten STR loci in Central Poland population]. AB - Allele frequency data for ten STR loci: D3S1358, vWA, D16S539, D2S1338, D8S1179, D21S11, D18S51, D19S433, TH01, FGA were estimated in a sample of 350 unrelated individuals in Central Poland population (Lodz region). The accordance with the Hardy and Weinberg equilibrium (HWE) for all investigated loci was proved. The allele distributions were compared to the similar data for other populations. PMID- 15129483 TI - [Determination of bromochlorodifluoromethane (Halon 1211) in biological material by gas chromatography with mass detector]. AB - A gas chromatographic method with mass spectrometry has been developed for the determination of bromochlorodifluoromethane (Halon 1211) in biological material (whole blood, organ samples). After incubation of the sample (temp. 65 degrees C, 15 min), 10 microliters of the headspace is analysed using a capillary column DB 5 ms (30 m x 0.25 mm x 0.25 micron). Quantitative analysis was made with the use of a single ion monitoring option--m/z 85 and m/z 87. This developed method was used to determine the concentration of bromochlorodifluoromethane in biological material collected from the body of the man whose death was due to intoxication of Halon 1211--fire-extinguisher contents. PMID- 15129484 TI - [Post mortem diffusion of carbon monoxide to muscles and blood--preliminary examinations]. AB - The purpose of the study was to determine whether the postmortem diffusion of carbon monoxide (CO) significantly affected the results of carboxyhemoglobin (COHb) and carboxymyoglobin (COMb) determinations. The musculocutaneous and muscular specimens collected from adult cadavers were used. The specimens were treated with CO for 24 h at room temperature. COHb and COMb were determined using gas chromatography. It was demonstrated that the skin substantially limited the diffusion of CO which slightly penetrated only the superficial layers of the muscle and did not change the blood level of COHb in the 4.5-cm layer of the muscle located underneath. The CO diffusion through the superficially charred and thermally coagulated muscle did not differ from that observed in the intact integuments. On the other hand, the membrane of the skin completely deprived of the adipose layer was not the barrier to moderate diffusion into the blood layer situated below. Thus, in charred corpses the results pf COHb and COMb determinations in the material collected under the layer of charred and coagulated tissues enable us to determine whether the victim was alive at the moment of the outbreak. PMID- 15129485 TI - [Burden of proof in medical cases--presumption of fact and prima facie evidence. 1. Burden of proof]. AB - The aim of this paper was to present the main rules concerning the burden of proof in polish civil trials, including medical cases. This paper also describes the subject of evidence were presented and explained. The court influence on evidence procedure was also analysed. The effect of the institution of informed consent on burden of proof in polish civil law is also described. This paper includes numerous High Court sentences on evidential and medical issues. PMID- 15129486 TI - [Burden of proof in medical cases--presumption of fact and prima facie evidence. II. Presumption of fact and prima facie evidence]. AB - The aim of this paper was to present the main rules concerning the burden of proof in polish civil trials, including medical cases. The standard rules were presented with all the important exclusions such as presumption of law and fact or prima facie evidence. The author analyses the effect of these institutions on burden of proof in medical cases. The difference between presumptions of fact and prima facie evidence was analysed and explained. This paper also describes the importance of the res ipsa loquitur rule in United Kingdom and USA. This paper includes numerous High Court sentences on evidential and medical issues. PMID- 15129487 TI - [Medical and legal aspects of inability to participate in legal proceedings and the execution of imprisonment resulting from mental illness]. AB - A mental disorder of the criminal can be the cause of reprieving penal conduct or the execution of imprisonment. In the present study the matter of ability to take part in legal proceedings and arrest in preliminary custody were considered. The possibility and propriety of staying in conditions of imprisonment mentally ill persons was also considered. Legal settlements relating to this problem were submitted. We paid attention to medical circumstances in which judicial experts should act in such cases. PMID- 15129489 TI - [Lethal injuries caused by sharp tools]. AB - Two interesting cases of fatal impactment on a sharp object were described. Firstly a fall through a glass door, buttocks first and piercing the iliac artery, and a case of falling onto a sharp handle of a pair of pliers carried in the pocket with the subsequent piercing of the heart. PMID- 15129488 TI - [Death attributed to toxic interaction of tramadol and other drugs]. AB - In this paper the authors presented the case of a 37-year old deceased investigated at the Department of Forensic Medicine of the Medical University of Gdansk. The subject of chemo-toxicological analysis were blood and urine samples as well as specimens collected from the liver, kidney and brain during autopsy. TLC, GC-FID, GC-MS were used as diagnostic tools. The determined tramadol concentration was 1.7 micrograms/ml in the blood and 15.5 micrograms/ml in the urine. PMID- 15129491 TI - Can we reawaken the spirit? PMID- 15129490 TI - Complementary medicine meets traditional system. AB - The Joint Commission's Continuum of Care Chapter addresses processes that apply to complementary care providers. PMID- 15129493 TI - Shrinking medication errors down to size. AB - Learn the best defense against medication errors--understanding the roles that human factors and systems play. PMID- 15129492 TI - Should you hire that nurse? AB - Negligent hiring, retention, and supervision practices expose you to liability. Test your knowledge with the following questions, then check your answers at http://www.nursingmanagement.com. PMID- 15129494 TI - Enhance end-of-life care. AB - A university-based veteran's medical center targets caregiver communication and patient feedback as the foundation of its end-of-life care quality improvement initiative. PMID- 15129495 TI - Protecting the older adult. AB - Learn the types of elder abuse plus risk factors, warning signs, and strategies to help your staff protect older patients. PMID- 15129496 TI - Professional presence: how do you get it? AB - Learn the six characteristics of professional presence and how to develop each trait. PMID- 15129497 TI - Under pressure. Weighed down by a high number of pressure ulcers, caregivers get a boost from a new initiative. AB - An evidence-based approach to treating pressure ulcers significantly improves key process and outcome indicators at this rural facility. PMID- 15129498 TI - How to build a cardiac surgery program. AB - Follow one hospital's experience in opening a new heart center, including its philosophy, triaging of issues, and staff preparation to achieve successful patient outcomes. PMID- 15129499 TI - Vascular access devices pinpoint safety. AB - Use the latest in vascular access products to comply with the Needlestick Safety and Prevention Act. PMID- 15129500 TI - Evidence for opioid variability, Part 1: A biological perspective. AB - The interplay of biological and pyschosocial factors explain the unique individual differences in opioid requirements that occur among postoperative patients. Nurses must be familiar with the physiologic mechanisms that influence opioid analgesia. Despite growing evidence that doses of opioid analgesics needed to achieve pain relief vary significantly from patient to patient, standardized dosing regimens continue to drive administration of medications for postoperative pain. In this article, evidence-based literature related to the biologic factors that contribute to differences in responses to opioid medication will be examined. Content will assist the Perioperative Nurse to recognize the pharmacology of opioid analgesics and the scientific basis for differences in the body's ability to metabolize and excrete opioids. These differences include age, gender, genetic predisposition, type of surgical procedure, preexisting pain, and prior or concurrent opioid use. Specific terms are introduced and defined to increase understanding of opioid variability. PMID- 15129501 TI - Transurethral microwave therapy of the prostate. AB - Benign prostatic hypertrophy (BPH) is a medical condition causing obstructive urinary symptoms that will inevitably affect the majority of the male population. Treatments for BPH have traditionally included invasive surgical intervention to resect a portion of the prostate as a relief of obstructive symptoms or medications that act in several ways to open the urinary channel and relieve symptoms. A new, less invasive treatment for BPH has now been developed and approved. Transurethral microwave therapy is discussed in this article. A case study is presented. PMID- 15129502 TI - Transperitoneal laparoscopic radical prostatectomy. AB - The diagnosis of prostate cancer for any male of any age can profoundly affect his life and that of his family. Improvements in laparoscopic devices, combined with associated surgical equipment and innovative urologic uses, have changed the treatment choices and outcomes for these types of patients. Transperitoneal laparoscopic radical prostatectomy (TLRP) offers improved postoperative outcomes for patients with localized prostatic cancer, decreasing the profound postoperative effects on functional return and potency. Quantitative advantages of TLRP include increased safety, lower hospital costs, and length of stay, while qualitative advantages include increased patient satisfaction, accelerated recovery, and qualitatively improved functional return. PMID- 15129503 TI - One institution's perspective on fast tracking same-day surgery patients. AB - This article describes a project for fast tracking of patients in one ambulatory surgical setting. Fast tracking may be a new endeavor or a well-entrenched practice in same-day surgery settings, depending on the organization. However, there is little information on this presently found in literature. Fast tracking is also known as post anesthesia care unit bypass. It allows stable patients that meet a set criteria to proceed, in this instance, from the operating room to the ambulatory procedure unit. This is a US government work. There are no restrictions on its use. PMID- 15129504 TI - Urinary incontinence in a vulnerable population: older women. AB - Urinary incontinence creates a tremendous hardship on older women and results in social isolation, anxiety, low self-esteem, depression, and decreased life satisfaction. Stress, urge, overflow, and functional incontinence can be treated by behavioral methods, medications, or invasive techniques. The nurse should complete a health history before making an informed decision with the older women about the most appropriate treatment plan. There can be improved quality of life for women with incontinence if health care providers focus on this problem. PMID- 15129505 TI - Assessment of the older female surgical patient. AB - Many older women enjoy better health than ever. Improved health care for women creates populations of older women who also have mental illness, chronic illness, are mentally impaired, or have dementia. Although aging itself may not be considered a surgical risk, those coexisting conditions predispose many older women to be vulnerable or at risk for a surgical procedure. A few of the major problems recounted by caregivers are reviewed here. Among them are disorientation, incontinence, and decubitus ulcers. PMID- 15129507 TI - Perioperative nursing data set for the adult patient having a cystoscopy. AB - This article gives a brief summary of a student project required in a perioperative internship program in a large medical center hospital. The Perioperative Nursing Data Set (PNDS) was used to determine activities necessary in developing a nursing care plan for patients undergoing cystoscopy. The project description and the total care plan is shared with the reader. PMID- 15129506 TI - Transurethral resection of the prostate syndrome. AB - Benign prostatic hypertrophy (BPH) is the most common benign tumor in men. It is a regional nodular growth of the prostate gland causing symptoms of urinary hesitancy and frequency. Transurethral resection of the prostate (TURP) is the definitive treatment for BPH. Absorption of large amounts of irrigating fluid can cause serious cardiovascular and central nervous system complications known as TURP syndrome. Close attention and assessment of the patient's vital signs and mental status by the registered nurse can increase the early detection and treatment of TURP syndrome. This is a US government work. There are no restrictions on its use. PMID- 15129508 TI - Percutaneous nephrostolithotomy: a means of managing urinary calculi. AB - Percutaneous nephrostolithotomy is a minimal invasive procedure designed to identify and remove kidney and ureteral stones using a percutaneous tract. Although a radiologist establishes the tract, the preoperative preparation and setup by the circulating nurse and surgical technician establishes appropriate equipment, a safe surgical environment for skin preparation, positioning, draping, and for performing the surgical procedure with a positive patient outcome. PMID- 15129510 TI - Eye of the beholder. AB - Three first semester nursing students collaborate to present their interpretations of their first observations in the operating room. This article includes a true story and topics addressing the operating room environment, the surgical team members, and the learning experience. PMID- 15129509 TI - A brief analysis of historical trends in nursing education: is there application to the current environment? AB - The history of nursing is circuitous and inconsistent; it has not always been based on scientific principles and delivered by providers with ethical and moral standing. Much of the image of nursing relates directly to the education, or lack of, over the course of its natural history. This article is a snapshot of the history of nursing education with application to the issues, risks, and changes of today. PMID- 15129511 TI - Perioperative nursing education: the operating room as a learning climate. AB - Perioperative patient care presents a demand for registered nurses with competence, skills, and experience. Without a well-grounded educational program, the novice perioperative nurse is often overwhelmed at the learning demands of the perioperative practice setting. Settings in which invasive and other operative procedures are carried out present a variety of challenges in orienting the novice nurse. There must be educational resources, coaching and support for novices who have limited real-world operating room experience, and work environments that encourage and foster professional development and a learning culture. An orientation program that focuses on the tenets of learning rather than seeing and doing without understanding holds the most promise for both recruitment and retention of perioperative nurses. PMID- 15129512 TI - The registered nurse first assistant classroom that isn't really a classroom. AB - Faculty and students in the 21st century need to be skilled in the use of computer technology. In this registered nurse first assistant (RNFA) program, distance learning modalities are used to link students and faculty from different locales, expanding the accessibility of RNFA education. Independent study is used to stimulate critical thinking and skill acquisition in a safe, user friendly environment. The strategies described are simple yet powerful and can be similarly developed by faculty in other RNFA programs without the need for large budgetary expenditures or sophisticated technology. PMID- 15129513 TI - Graduate education: the perioperative nurse practitioner. AB - As the surgical experience becomes increasingly complex, the need for nurses who are experienced and educated in the management of surgical problems becomes critical. This article discusses the development of a perioperative nurse practitioner program designed to prepare perioperative nurses to assume the complex and exciting role of caring for a diverse population of surgical patients. PMID- 15129514 TI - What's your specialty, Doc? Perioperative nursing. AB - Perioperative nurses who are contemplating the pursuit of a doctoral degree in nursing need information about the current status and characteristics of doctoral education to make informed decisions. This article addresses questions both philosophic and pragmatic that should be addressed by prospective students. Information sources are identified for assistance with making the selection from the 75 plus programs that offer a nursing doctorate nationwide. Suggested sources include professional organizations as well as current students, faculty, and alumni. Determinations should be based on academic information, such as similarities and differences in degrees awarded; the program of studies leading to the degree; admission and progression standards; research interest of the student relative to identified focus and expertise of program faculty, dissertation process, and university and nursing education resources and services, including financial and technological support. Attainment of the doctorate is presented as an enriching experience that prepares nurses for leadership in both education and health care as clinicians, educators, and administrators. Particular attention should be paid to selecting programs that compare favorably with the published American Association of Colleges of Nursing quality indicators for doctoral programs. There must be strong support in the program from both faculty mentors and peer colleagues, reflective of a community of scholars, pursuing not only individual but also collective wisdom, whereas engaging an individual's mind and soul in the pursuit of new knowledge and skills in an atmosphere of mutual trust and respect. PMID- 15129515 TI - Certification: perspectives on competency assurance. AB - As registered professional nurses, we have an obligation to our patients to assure we are practicing at a competent level. Nurse managers and administrators must show to the Joint Commission for Accreditation of Healthcare Organization (JCAHO) that the nurses providing care in their facility are competent. Assessing competency is not a simple task, and there is no 1 correct answer or definition of competence or competency assessment. Some of the methods available for competency assessment are self-assessment models, multiple choice examinations used for certification and licensure, oral or essay examinations, and practice or patient simulations. The Certification Board Perioperative Nursing (CBPN) recognizes that learning and competency maintenance is accomplished in a variety of ways, and that learning is achieved through successful participation in various professional activities. Becoming certified and remaining certified, or completion of the Professional Perioperative Portfolio are ways for nurses to continue to show competency in perioperative nursing. Research is vital to validate that certification does indeed show competency. The research committee of CBPN and the Nurse Credentialing Research Coalition (NCRC) are committed to quantify not only the value of the credential but to examine the personal, professional, and practice outcomes of certified nurses across the country. It is hoped that the current research being conducted will eventually allow concerned parties to have assurance that the certified professional registered nurse is indeed a competent registered nurse. PMID- 15129516 TI - Teaching an online course. AB - An online pharmacology course was developed for nursing students in the associate degree program at Delaware County Community College, Media, PA. The course was a response to an expressed need from students for more pharmacology content in their curriculum and as part of an articulation agreement with Widener University School of Nursing, Chester, PA, registered nurse to a masters of science in nursing program. Course design was a team effort among faculty and instructional technology specialists. Faculty developed a list of features the course needed to deliver the content as well as instructional activities designed to help learning to occur. Instructional technology specialists designed the course layout, chose the appropriate software needed to support the course design, and developed original programming as necessary. These elements were woven into a course that is tailored to our instructional design. In the years since the course was first offered, changes have been made to increase interaction between students and to provide all aspects of the course online, including testing. Teaching online challenges faculty to learn new technical skills as well as to think about new ways of teaching. PMID- 15129517 TI - Assessment techniques for web-based instruction: lessons learned from teaching a graduate course in instructional technology. AB - In an attempt to discover how students learn in an online learning environment, as well as to investigate how online communication patterns differ from those in a face-to-face class, the authors used students participating in a graduate program in instructional technology in Philadelphia, PA. Various activities and assignments were used to elicit information about student learning with, and attitudes toward an online learning environment. Over a period of 3 semesters, student data were collected through qualitative and quantitative methods, including observations, surveys, tests, in-class discussions, and e-mail communication analysis. Results for these assessment methods are included in this article, as well as implications for future investigation into this topic. PMID- 15129518 TI - Education in practice: the United Kingdom experience. AB - Health care leaders, government agencies, and professional nursing associations all understand that rethinking existing delivery strategies is part of the strategic approach to workforce planning. In the United Kingdom, the demand for nurses is great and the supply inadequate. The potential for a public health crisis is challenging the way the United Kingdom educates and positions nurses within the health care industry. This article reviews past efforts and current strategies in workforce characteristics, age, educational mix, competencies, career goals, and future plans to ensure adequate nurses to care for the health needs of the people residing in the United Kingdom. The activities of the National Association of Theatre Nurses in promoting dialogue around this issue and fostering collaborative initiatives are presented in a framework of efforts designed to benefit all. PMID- 15129519 TI - The registered nurse as a first assistant: the "downunder" experience. AB - In 1996, Bernadette Brennan, RN, was awarded a fellowship from the Australian Confederation of Operating Room Nurses (now known as the Australian College of Operating Room Nurses Ltd.) to travel to the United States to study the role of the Advanced Nurse Practitioner, with an emphasis on the role of the registered nurse first assistant (RNFA). As part of this study, she undertook an RNFA course at Delaware County Community College. This article provides a description of her work to develop an educational program for RNFAs in Australia. Because of the size of the country and the many rural areas needing to be served, her challenge was to devise an accessible program that was also academically rigorous. Her trials as well as her triumphs are presented here. PMID- 15129520 TI - Patient safety, errors and mistakes, and perioperative nursing. AB - Today's world of advancing technology in health care represents complex diagnostic, operational, and administrative processes, all of which must be coordinated to ensure the delivery of safe, quality health care. Nonetheless, the health care industry's implementation and practice of quality measures fall below the standards of other industries, such as aerospace, telecommunications, or information technology. With health care performing at this lower level, it is no wonder that headline grabbers dwell on the deaths that occur as a result of medical errors. A new emphasis on quality management is essential in nursing education and in quality management to improve US health care. This article summarizes recent reports on medical errors and suggests strategies to improve patient safety. PMID- 15129521 TI - Fill 'er up! PMID- 15129522 TI - Information management takes center stage. AB - Recognizing information management as a critical issue, the Joint Commission has deemed it a core chapter in all of its manuals. PMID- 15129523 TI - Budget training: it's overdue. AB - Learn frequently used terminology, suggested steps, and goals to preparing your budget. PMID- 15129524 TI - What's the difference? PMID- 15129525 TI - Competent i.v. management, Part 2. AB - Implementation of fundamental principles and practice standards regarding peripheral i.v. therapy reduces the risks of patient injury and litigation. PMID- 15129526 TI - Skin tears. AB - Skin tears are a serious, painful problem for older patients. Find out how your staff can recognize patients at risk, what they can do to prevent skin tears, and how to manage them effectively if they occur. PMID- 15129527 TI - www.alternativecasemanagement.now. AB - Nurse case managers at this integrated delivery system implement an alternative, Web-based case management approach. PMID- 15129528 TI - Out of a partnership emerges an internship. An advantageous alliance. AB - A university-based RN BS program and a hospital-based diploma school of nursing partner to provide a clinical internship for RNs. PMID- 15129529 TI - Should continuous lateral rotation therapy replace manual turning? AB - Learn the pulmonary complications of immobility in the critically ill, which are associated with significant morbidity and mortality. PMID- 15129530 TI - ED wonders. AB - Review what's new in biphasic defibrillation and trauma nursing, including and intubation alternatives. PMID- 15129531 TI - Humanitarian aspects of health support in operations other than war. AB - The decade of the 1990s proved to be very busy for health personnel serving in the Australian Defence Force (ADF). During this time, they deployed to areas as far away as Africa, after the genocide in 1994, and areas closer to home. There have been missions in Indonesia, Bouganville, and Papua New Guinea. In more current times, the most significant deployment has been to East Timor where health personnel remain today. Furthermore, there have been a number of deployments on short missions to various areas within Australia's strategic area of interest. Every mission has a mission statement. In all cases involving health personnel, the statement outlines their primary tasking, which will be the provision of health support to the troops in force. The secondary tasking, which inevitably follows, is the provision of health support to the local community from within any spare capacity. It is this spare capacity that this article will discuss. The primary focuses of this article are what care should be provided and the limitation of such care, how health care should be provided, what training is required for health personnel who will provide this care, and the policies that are required to be in place to monitor this care. PMID- 15129532 TI - Intensive care nursing in Rwanda, 1994-1995. AB - The deployment of Australian Military personnel in support of the United Nations Aid Mission in Rwanda (UNAMIR II) in August 1994 represented the first large contingent of Australian medical personnel since the Vietnam War. A kaleidoscope of emotions filled the minds of many deploying personnel before, during and after the deployment. Yet through this turmoil, there were many valued lessons learned. For many, it was a reality check--a return to the basics of life. This article is a personal account of the experience of an intensive care nurse working in a third-world country performing first-world medicine. PMID- 15129533 TI - Rwanda: gods and ghosts. AB - This paper provides a personal story. I was born in Africa and have a great empathy for its people. The perceptions I provide are through the eyes of an African born individual who has lived in Australia for many years. It is said that you can take the child out of Africa; however, you cannot take Africa out of the child. During my return to Rwanda in 1994 to 1995, I learned many lessons. Perhaps the most significant being that the Rwandan people displayed absolute stoicism and acceptance of adversity, and from the suffering they had endured, they displayed a great dignity. PMID- 15129534 TI - Australian military trauma management: experiences in Rwanda and Papua New Guinea. AB - The military trauma management experiences in the 1950s and 1960s were some of the catalysts for the development of emergency medical services systems worldwide. How is the Australian military managing trauma in light of the recent experiences in Rwanda and Papua New Guinea? PMID- 15129536 TI - Return to Bangka Island. AB - This article is a return in a couple of ways to one of the most tragic events in the history of Australian military nursing. Firstly, it describes how the evacuation of nurses from Singapore in 1941 led to circumstances that resulted in the massacre or internment of many of those women. Then in 1993, a group of surviving World War II nurses and current serving Australian Army nurses returned to the site of their sorrow. PMID- 15129535 TI - Composition and performance of a small resuscitation team in a deployed military hospital. AB - The Royal Australian Navy deployed a 5-person resuscitation team, including 2 nursing officers, to East Timor from February to August 2000 as part of the United Nations Military Hospital. The team managed a wide variety of emergency presentations, effectively utilising the Advanced Trauma Life Support (ATLS) model. The 2 nursing officers, in addition to using their clinical skills, also provided leadership, enhancing communication and cohesion within the team and ultimately providing an essential contribution to the team's excellent patient outcomes. PMID- 15129537 TI - What to do before disaster strikes. AB - The key to averting disaster or minimizing its effects is a solid data security plan. Learn the types of data risks that exist, how to develop a plan to counteract them, and your role in ensuring the safety of health and patient data. PMID- 15129538 TI - Coach staff to initiate change. AB - Empower staff to get involved with changes to patient practices and outcomes. PMID- 15129539 TI - Lead outside your comfort zone. AB - Successful leaders use their passion, values, and personal mission to create and maintain influence. Learn to define and develop a context that allows you to successfully lead nursing and non-nursing departments. PMID- 15129540 TI - The UAP dilemma. AB - Although studies show adverse patient outcomes with increased use of unlicensed assistive personnel (UAPs), improvement in their training and an appropriate staff mix may assure quality care. PMID- 15129541 TI - Coming of age. Cystic fibrosis--once a childhood disease--makes its way to adulthood. AB - Cystic fibrosis patients and their caregivers face unusual complexities when these patients move from the pediatric to adult setting. PMID- 15129542 TI - How to make conflict work for you. AB - Fostering an environment of collaboration allows staff room to solve conflicts among themselves. PMID- 15129543 TI - A closer look at chemotherapy errors. AB - Lehigh Valley Hospital and Health Network, a 700-bed tertiary care system, created a reference guide as part of its chemotherapy error prevention initiative. PMID- 15129545 TI - NAs aid RN retention. PMID- 15129544 TI - Selling points. PMID- 15129546 TI - Trust me or not? PMID- 15129547 TI - Within reach. AB - Learn how one rural community hospital overcame staffing and funding shortages to create a successful, collaborative orientation program for its critical care nurses. PMID- 15129548 TI - RATs improve heart care. PMID- 15129549 TI - Medication errors: a lesson from long-term care. PMID- 15129551 TI - The metabolic rate constants and specific activity of human and rat hepatic cytochrome P-450 2E1 toward toluene and chloroform. AB - Chloroform (CHCl3) is a near-ubiquitous environmental contaminant, a by-product of the disinfection of drinking water sources and a commercially important compound. Standards for safe exposure have been established based on information defining its toxicity, which is mediated by metabolites. The metabolism of CHCl3 is via cytochrome P-450 2E1 (CYP2E1)-mediated oxidation to phosgene, which is known to obey a saturable mechanism. CYP2E1 is a highly conserved form, expressed in all mammalian systems studied, and is responsible for the metabolism of a great many low-molecular-weight (halogenated) compounds. However, the Michaelis Menten rate constants for CHCl3 oxidation have not been derived in vitro, and the specific activity of CYP2E1 toward CHCl3 has not been reported. In this investigation with microsomal protein (MSP), apparent Vmax values of 27.6 and 28.3 nmol/h/mg MSP and apparent K(m) values of 1 and 0.15 microM in rats and human organ donors, respectively, were demonstrated. The specific activity of CYP2E1 toward CHCl3 in rats and humans was 5.29 and 5.24 pmol/min/pmol CYP2E1, respectively. Toluene metabolism to benzyl alcohol (BA), another CYP2E1-dependent reaction, was also highly dependent on CYP2E1 content in humans, and was more efficient than was CHCl3 metabolism. The specific activity of human CYP2E1 toward toluene metabolism in human MSP was 23 pmol/min/pmol CYP2E1. These results demonstrate that differences in CYP2E1 content of MSP among individuals and between species are largely responsible for observed differences in toluene and CHCl3 metabolism in vitro. PMID- 15129550 TI - Development of a physiologically based pharmacokinetic model for chlorobenzene in F-344 rats. AB - A physiologically based pharmacokinetic (PBPK) model to describe the absorption, distribution, metabolism, and elimination of chlorobenzene in rats was developed. Partition coefficients were experimentally determined in rat tissues and blood samples using an in vitro vial equilibration technique. These solubility ratios were in agreement with previous reports. The in vivo metabolism of chlorobenzene was evaluated using groups of three F344 male rats exposed to initial chlorobenzene concentrations ranging from 82 to 6750 ppm in a closed, recirculating gas uptake system. An optimal fit of the family of uptake curves was obtained by adjusting Michaelis-Menten metabolic constants, K(m) (affinity) and Vmax (capacity), using the PBPK model. At the highest chamber concentration, the uptake curve could not be modeled without the addition of a first-order (Kfo) metabolic pathway. Pretreatment with pyrazole, an inhibitor of oxidative microsomal metabolism, had no impact on the slope of the uptake curve. The completed PBPK model was evaluated against real-time exhaled breath data collected from rats receiving either an intraperitoneal (i.p.) injection or oral gavage dose of chlorobenzene in corn oil. Exhaled breath profiles were evaluated and absorption rates were determined. Development of the chlorobenzene PBPK model in rats is the first step toward future extrapolations to apply to humans. PMID- 15129552 TI - Does high organochlorine (OC) exposure impair the resistance to infection in polar bears (Ursus maritimus)? Part I: Effect of OCs on the humoral immunity. AB - This study was undertaken to assess if high levels of organochlorines (OCs) are associated with decreased ability to produce antibodies in free-ranging polar bears (Ursus maritimus) and thus affect the humoral immunity. In 1998 and 1999, 26 and 30 polar bears from Svalbard, Norway, and Churchill, Canada, respectively, were recaptured 32-40 d following immunization with inactivated influenza virus, reovirus, and herpes virus and tetanus toxoid. Blood was sampled at immunization and at recapture for determination of plasma levels of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs), serum immunoglobulin G (IgG) concentrations, and specific antibodies against influenza virus, reovirus, and herpes virus, tetanus toxoid, and Mannheimia haemolytica. The OCs alone contributed with up to 7% to the variations in the immunological parameters. The combination of sigma PCBs (sum of 12 individual PCB congeners), sigma OCPs (sum of 6 OCPs), and biological factors accounted for 40-60% of the variation in the immunological parameters. Negative associations were found between sigma PCBs and serum immunoglobulin G (IgG) levels and between sigma PCBs and increased antibody titers against influenza virus and reovirus following immunization. In contrast, a positive association was registered between sigma PCBs and increased antibodies against tetanus toxoid. sigma OCPs also contributed significantly to the variations in the immunological responses. OCs did not have the same impact on the antibody production against M. haemolytica. The present study demonstrated that high OC levels may impair the polar bears ability to produce antibodies and thus may produce impaired resistance to infections. PMID- 15129553 TI - Modulation of fatty acid methyl esters in rats pretreated with tri-o-tolyl phosphate. AB - Formation and toxicity of fatty acid methyl esters (FAMEs) have been reported both in vitro and in vivo. In previous studies, it was shown that fatty acid ethyl ester synthase (FAEES), which catalyzes the formation of FAMEs, also expresses esterase activity. Therefore, it was hypothesized that inhibitors of esterases such as tri-o-tolyl phosphate (TOTP) can modulate the formation of FAMEs. To test this, four groups of rats were used. Group 1 served as control (vehicle only). Group 2 was treated with methanol only (3 g/kg via gavage), group 3 was given TOTP only (100 mg/kg i.p. in corn oil), and group 4 was administered TOTP as in group 3, followed by methanol after 18 h. Three hours after exposure, animals were sacrificed and FAEES activity and FAME levels were measured in blood, liver, pancreas, and brown fat. About 95% of FAEES activity was inhibited in the liver and whole blood of TOTP-treated rats (group 3) but no inhibition was observed in the pancreas or brown fat. Total hepatic FAMEs were found to be lowest for the TOTP-treated group (3) and highest in the methanol-treated animals (group 2). Total pancreatic FAMEs in different groups were not statistically different, while significant increases were observed in the brown fat in both methanol-treated groups. To verify that the oxidative metabolism of methanol was unaffected by TOTP, alcohol dehydrogenase activity was also measured and found to be unchanged in any group as compared to control. These results demonstrate that the formation of FAMEs can be modulated in the liver and probably in blood, but not in the pancreas or brown fat by the inhibitors of FAEES. PMID- 15129555 TI - [Oral surgery in general dental practice 1. Preface]. AB - Delegation of basic dental treatment to well-trained oral hygienists, dental nurses, and dental technicians will result in more time for the general practitioner to treat more specialized dental problems. In two issues of this journal, some surgical treatments which can be delivered by a dentist with special skills on this topic, are discussed. This first issue concentrates on the treatment of odontogenic abscesses, the removal of teeth and roots, the treatment of a perforation of the maxillary sinus floor, and apectomies. PMID- 15129554 TI - Green tea protection against age-dependent ethanol-induced oxidative stress. AB - Ethanol intoxication leads to oxidative stress, which may be additionally enhanced by aging. The aim of this study was to investigate the influence of green tea as a source of water-soluble antioxidants on the ability to prevent oxidative stress in aged rats sub-chronically intoxicated with ethanol. Two-, 12 , and 24-mo-old male Wistar rats were divided into 4 experimental groups: (1) control, (2) green tea, (3) ethanol, and (4) ethanol and green tea. Ethanol intoxication produced age-dependent decrease in the activity of serum superoxide dismutase, glutathione peroxidase, and reductase and in levels of glutathione (GSH), vitamins C, E, and A, and beta-carotene. Changes in the serum antioxidative ability were accompanied by enhanced oxidative modification of lipid (increase in lipid hydroperoxides, malondiadehyde, and 4-hydroxynonenal levels) and protein (rise in carbonyl group levels). Green tea partially protected against changes in antioxidant enzymatic as well as nonenzymatic parameters produced by ethanol and enhanced by aging. Administration of green tea significantly protects cellular components such as lipids and proteins against oxidative modification. Results indicate that green tea effectively protects blood serum against oxidative stress produced by ethanol as well as aging. PMID- 15129556 TI - [The odontogenic abscess. Aetiology, treatment and involvement in the orofacial region]. AB - Odontogenic infections are a common problem in daily practice. Occasionally, an odontogenic infection evolves an abscess. This article discusses the aetiology, the treatment and the involvement of odontogenic abscesses in the oro-facial region. Their occurrence, course and treatment are depending on the patient's immune response, and on microbial and environmental factors. PMID- 15129558 TI - [Dentoalveolar surgery for the dentist: removal of third molar]. AB - In contrast to removal of other teeth and roottips, a third molar is mostly removed for preventive reasons. There is still debate about the correct indications for removal of third molars. As soon as the decision to remove a third molar surgically is made, the dentist has to decide between performing the surgical procedure himself or referring the patient to an oral and maxillofacial surgeon. Level of difficulty of the treatment and experience, available time, availability for postoperative care, and personal interest of the dentist are issues influencing this decision. This article describes systematically the indications, for instance using preoperative radiodiagnostics, the factors determining the technical surgical plan, as well as the practical surgical procedures. PMID- 15129557 TI - [Dentoalveolar surgery for the dentist: removal of teeth and root tips]. AB - The indication for surgical removal of teeth or roottips is often made in dental practice. In some cases a general practitioner will decide to perform the surgical procedure himself, while in other cases he will refer the patient to an oral and maxillofacial surgeon. Level of difficulty of the treatment and the experience, the time available, the availability for postoperative care, and the personal interest of the dentist are factors involved in decision making. It is likely that with increased experience, the dentist will be able to perform more complicated treatments. This article supports this process. Surgical removal of teeth and roottips is systematically described, with emphasis on technical aspects. Presurgical management, removal of singlerooted and multirooted teeth, woundcare and postoperative management are the subjects covered. PMID- 15129559 TI - [Oroantral perforations]. AB - Some risk factors in relation to the occurrence of an oroantral perforation are discussed in the light of the existing literature. Factors relevant for the diagnosis and the treatment of an oroantral perforation, are discussed. Small perforations probably heal without problems, provided that the alveolus is deep enough and a blood clot will form without disturbance. Some pros and cons of buccal and palatal transposition flaps for closing clinically significant oroantral perforations, are discussed. PMID- 15129560 TI - [Apical endodontic surgery]. AB - If (a revision of) a conventional endodontic treatment is not possible or not successful, apical endodontic surgery can be indicated. The contemporary indications, the better retrograde preparation techniques with ultrasonic retro tips, and the better visualisation of the operation area with an operation microscope can lead to higher success percentages. Moreover, the current developments in the field of compatible filling materials are promising. Also the application of lasers is promising, but has still to prove its clinical usefulness. PMID- 15129561 TI - [Dental anxiety]. PMID- 15129562 TI - [Diabetes mellitus, but which type?]. AB - In three patients with an unusual presentation of diabetes mellitus, the classification of their diabetes was troublesome. An adolescent male with slightly elevated blood-glucose levels turned out to have excellent glycaemic control on sulphonylurea derivatives only. When he was 40 years of age, his diabetes was finally diagnosed as 'maturity onset diabetes of the young' (MODY). A non-obese 41-year-old man was initially diagnosed with type-2 diabetes. Therapy with oral hypoglycaemic agents was unsuccessful and he was subsequently classified as having 'latent autoimmune diabetes of adults' (LADA) based on the presence of antibodies against glutaminic acid decarboxylase. A 29-year-old man presented with severe ketoacidosis and was initially believed to have type-1 diabetes. The patient himself discontinued insulin therapy and he was eventually diagnosed as having type-2 diabetes. A careful classification may have clinical consequences. Patients with MODY3, for example, respond to sulphonylurea derivatives. In MODY2, treatment with diet alone is often sufficient. In patients with LADA, insulin therapy is the treatment of choice. The recognition of diabetes mellitus type 2 as the underlying illness in some patients who present with ketoacidosis means that these patients can be specifically treated for their basic problem, which is insulin resistance. For them, weight reduction is essential and metformin is the drug of choice as far as pharmacotherapy is concerned, but of course attention must also be given to cardiovascular risk factors such as hypertension and dyslipidaemia. PMID- 15129563 TI - [Sexual contact between doctor and patient]. AB - Codes of medical practice forbid a doctor to have sex with a patient. Still, practice and attitudes turn out to be unruly, so there is a need for (public) discussion and education. Moreover, other professional limitations in the doctor patient relationship (e.g., 'non-medical' physical contact, dating a patient, accepting gifts from a patient, and treating partners, family-members and friends) require more attention to facilitate the reverification of the impermissibility of a sexual relationship with a patient. The need for such reverification is underlined by the expected changes in the balance between engagement and detachment in the doctor-patient relationship due to the influences on medical practice of technology, protocols and the discontinuity of personal care. PMID- 15129564 TI - [Toxicological aspects and health risks associated with hydroquinone in skin bleaching formula]. AB - The use of hydroquinone as a cosmetic skin-bleaching agent has been forbidden since January 2001. It is now available only on prescription. The ban has been introduced because of medium-term effects such as white patches on the skin, particularly on the face (leukoderma with confetti-like depigmentation), and subcutaneous dark collections of pigment (exogenous ochonosis). Long-term effects are a possibility; cancer being the most likely. Renal adenomas and leukaemia occurred in animal experiments indicating the nephrotoxicity and carcinogenic properties of the substance. It is now known how hydroquinone and its metabolites can cause damage to DNA and inhibit apoptosis of mutated cells. The carcinogenic action of benzene is difficult to attribute to its hydroquinone metabolite. Daily use of hydroquinone causes it to accumulate in the body as absorption into the skin is faster than excretion in the urine. The use of hydroquinone as a skin bleaching agent is accordingly unsafe and should be completely banned. Alternatives such as azaleic acid and thioctic acid (alpha-lipoic acid) are available. PMID- 15129565 TI - [Clinical reasoning and decision making in practice. A depressive foreign woman with symptoms of malaise]. AB - A 27-year-old woman was admitted to the hospital with a depression, anaemia and fatigue. She had come from Angola to the Netherlands as a refugee 2 years before this evaluation. As an explanation for her symptoms tropical infectious diseases of parasitic origin were considered, but no clues were found in this direction. The test for trypanosomiasis was considered to be suggestive for an infection in the past (persistent titre 1:200). She was discharged but readmitted 6 months later because of a deterioration of her clinical condition. Magnetic resonance imaging showed bilateral signal abnormalities within the white matter of the brain. On examination no neurological signs or abnormalities were found. Again, no definite diagnosis could be made and the patient was discharged. Because of a further deterioration of her clinical condition she was readmitted a short time later for the third time. On the MRI the white matter lesions had increased. The serum protein electrophoresis was markedly abnormal with an elevated IgM Level. Finally, at a repeated lumbar puncture mobile trypanosomes were found. The diagnosis of 'West African sleeping sickness' was made and the patient was treated with eflornithine. She recovered completely during the next 18 months. PMID- 15129566 TI - [Diagnostic image (185). A man with a pimple on the upper lip. Carbuncle of the upper lip]. AB - A 24-year-old man presented with a carbuncle of the upper lip. Surgical drainage was installed and flucloxacillin was administered. Full recovery ensued. In culture Staphylococcus aureus was abundant. PMID- 15129567 TI - [Sexual contact between general practitioner and patient in the Netherlands: prevalence and risk factors]. AB - OBJECTIVE: To obtain data on sexual contact between general practitioners (GPs) and their patients and to determine the relationship between sexual abuse and GP characteristics. DESIGN: Cross-sectional. METHOD: In February-May 2002, anonymous questionnaires were mailed to a randomized sample of 1250 general practitioners in the Netherlands. RESULTS: A total of 977 general practitioners responded (response: 80%). This number included relatively large numbers of women and GPs aged > or = 50. Out of the 977 GPs, 32 had had sexual contact with a patient at some time: 30/695 (4.3%) of the male GPs and 2/247 (0.8%) of the female GPs Coitus was reported by 24 (75%) of them. Of the 32 perpetrators 11 (34%) had had sexual contact with 2 or more patients. Of the 30 male GPs who engaged in sexual contact with patients, 20 (67%) afterwards felt positive about the sexual encounter for himself and 22 (73%) for the patient. Sexual contact with a patient occurred more often among men who at the time of the survey were aged < or = 50 [corrected]. The incidence was not related to the degree of urbanisation of the practice area or to the size of the practice partnership. CONCLUSION: Sexual contacts between general practitioners and their patients are not just incidents. It is not enough to be conscious of one's own sexual feelings towards patients. Implementation of a clear policy is needed. PMID- 15129568 TI - [Three patients with orf (ecthyma contagiosum)]. AB - Orf was diagnosed in three patients: a 16-year-old Moroccan girl who had cut her finger in a butcher's shop, a 47-year-old Dutch woman who had allowed a lamb to suck on her finger on a children's farm, and a 50-year-old Dutch farm woman. Orf or ecthyma contagiosum is a well-known viral disease among sheep and goats. Transmission to humans as a zoonosis is rare but can take place via direct contact with infected animals or animal products. The clinical picture is usually characterized by a solitary lesion that develops on the dorsal side of the fingers or hands. This viral condition produces little or no systemic complaints and the lesions usually regress spontaneously without scar formation within 6 weeks (range 4-9 weeks). The correct diagnosis can usually be made on clinical grounds. The diagnosis may be confirmed by demonstration of the virus by electron microscopy or the polymerase chain reaction in fluid obtained from the skin lesions or by conventional histopathology. Early clinical recognition and knowledge of this benign, self-limiting viral condition is vital to avoid unnecessary surgical intervention. Proper information and reassurance of the infected patient are very important. All three patients recovered. PMID- 15129569 TI - [Extreme dietary selectivity following liver transplantation in a 5-year-old girl; management via behavior therapy]. AB - In a 5.1-year-old girl who had been treated by surgical correction of biliary atresia and total orthotopic liver transplantation, extreme dietary selectivity was noted; this was treated by behaviour therapy. On entry in the rehabilitation centre, she manifested malnutrition along with a variety of gastro-enterologic complaints. The treatment consisted of a set of behavioural procedures such as stepwise expansion of the diet, verbal prompting, intermittent contingent attention and a list of agreements with a system of rewards; this led to the elimination of the dietary selectivity and to the consumption of a varied diet in a normal tempo. PMID- 15129570 TI - [Post mortem semen acquisition: not without 'informed consent']. PMID- 15129571 TI - [Systemic reviews as a basis for guidelines]. PMID- 15129572 TI - [Treatment of the obstructive sleep-apnea syndrome in adults]. PMID- 15129573 TI - [Primary adenocarcinoma of the small intestine]. PMID- 15129574 TI - [A survey of some new pharmaceutical specialties introduced in Belgium in 2003]. PMID- 15129575 TI - [Current advances in hemostatic and antithrombotic agents]. PMID- 15129576 TI - [Hemostasis and blood coagulation revisited: mechanisms and therapeutic implications]. AB - The number of oral anti-thrombotic agents is steadily expanding. Major advances have recently been made in the understanding of haemostasis and blood coagulation processes. This paper reviews the mechanisms of haemostasis and thrombosis and the molecules used for the treatment of arterial and venous thrombotic disorders with a special emphasis on recently introduced agents (Clopidogrel, Pentasaccharide, Melagatran). PMID- 15129577 TI - [Coagulation factors and hemostatic agents]. AB - A large variety of therapeutic agents are available to prevent or treat hemorrhagic events. An etiologic focusing is essential to permit a rational use of these drugs. Some medications are cheap, others are very expensive, and the clinician has to evaluate the cost-effectiveness of his prescription in each situation. PMID- 15129578 TI - [Medical treatment for cerebral atherothrombosis]. AB - Atherothrombotic ischemic stroke is associated with multiple vascular risk factors, including mainly hypertension, diabetes, and hyperlipidemia. Medical therapy of cerebral atherothrombosis implies control of these vascular risk factors and antithrombotic drugs as well. This paper reviews the current therapeutic guidelines according to the randomized trials for primary and secondary stroke prevention. PMID- 15129579 TI - [Risk of thromboembolism in contraception and hormone substitution: an update and recommendations]. AB - Venous and arterial thromboembolism events are unfrequent but classical complications of sex steroid therapy, in contraception and menopause. The relative risks induced by these treatments have recently highlighted by the general press, in particular for the 2nd and 3rd generation progestins in oral contraception. In this article, we will review the risks of venous and arterial thromboembolism in patients treated with oral contraception or postmenopausal hormone replacement therapy. Practical guidelines will be defined. PMID- 15129580 TI - Oedematous skin disease of buffalo in Egypt. AB - This review covers a historical view and etiology of oedematous skin disease which affects buffalo in Egypt, the microbiology of Corynebacterium pseudotuberculosis causing the disease: its virulence; clinical signs; mechanism of pathogenesis; histopathology; mode of transmission; immunological aspects; treatment and control. It is concluded that C. pseudotuberculosis serotype II is the main cause of OSD and exotoxin phospholipase D and its lipid contents of the cell wall are the major causes of pathogenesis. After declaring the role of Hippobosca equina in transmission of the causative agent among buffaloes, control of OSD is now available. PMID- 15129581 TI - Changes in lymphocyte subsets in the bronchus-associated lymphoid tissue of goats naturally infected with different Mycoplasma species. AB - The distribution of cells containing lysozyme, S-100 protein, CD3, CD4, CD8, major histocompatibility complex class II antigen and immunoglobulin G (IgG) was analysed in the bronchus-associated lymphoid tissue (BALT) of goats naturally infected with three Mycoplasma species. This study included the immunohistochemical characterization of the pneumonic lesions of 18 goats (3-5 months old) infected with one of the following Mycoplasma species: M. mycoides ssp. mycoides, Large Colony type (goats no. 1-6), M. mycoides ssp. capri (goats no. 7-12) and M. capricolum ssp. capricolum (goats no. 13-18). Microscopically, infected animals showed a moderate broncho-interstitial pneumonia, characterized by lymphoid hyperplasia of the BALT and infiltration of mononuclear cells in the alveolar walls and airways. The main cellular type in the BALT was represented by CD3+ T lymphocytes, and the ratio of CD4+:CD8+ cells was > 2. The BALT showed large germinal centres mainly composed of IgG+ B lymphocytes, with numerous S 100+ follicular dendritic cells. The presence of follicular dendritic cells confirmed the high degree of organization of this lymphoid tissue. The immunohistochemical results showed that activated T lymphocytes, particularly in the CD4 subset, and IgG+ B cells, play a major role in the immune response of the caprine lung infected with these species of mycoplasmas. PMID- 15129582 TI - Chronic fatigue and immune dysfunction syndrome associated with Staphylococcus spp. bacteraemia responsive to thiacetarsamide sodium in eight birds of prey. AB - Chronic fatigue and immune dysfunction syndrome (CFIDS) is a recognized human illness with zoonotic implications that is rarely described in animals. Eight birds of prey examined between 1992 and 1995 and sharing common symptoms (asthenia, inability to fly, poor appetite and emaciation) underwent laboratory tests revealing immunodeficiency, anaemia, high creatine kinase levels and low serum magnesium levels. Diagnosis of CFIDS was based upon these features. The effectiveness of an arsenic-based medication, thiacetarsamide sodium, administered intravenously for 2-3 days at low dosages (0.1 ml/kg/day) has been demonstrated by checks carried out 10, 20 and 30 days after therapy. The symptoms and the immune and haematological dysfunctions disappeared within 2-4 weeks of treatment. In all patients, micrococcus-like organisms found adhering to the outer surface of many red blood cells, had disappeared at post-treatment controls. Two of five blood cultures were positive for Staphylococcus spp. (S. intermedius and S. xilosus). Consideration is given to the pharmacological activity of an arsenic-based drug in animal illnesses resembling CFIDS. PMID- 15129583 TI - Clinical effects of experimental dual infections with porcine reproductive and respiratory syndrome virus followed by swine influenza virus in conventional and colostrum-deprived pigs. AB - Previous studies demonstrated that experimental dual infections of pigs with porcine reproductive and respiratory syndrome virus (PRRSV) followed by H1N1 influenza virus cause more severe disease and growth retardation than the respective single virus infections. Here three experiments were undertaken to better define the clinical impact of combined PRRSV-H1N1 infections in conventional and caesarean-derived colostrum-deprived (CDCD) pigs. Groups of pigs were inoculated by aerosol with PRRSV followed by H1N1 at 3-, 7- or 14-day intervals. During the post-H1N1 period, mean body temperatures, respiratory signs and mean weight gains in the PRRSV-H1N1 inoculated groups were recorded and compared with those in uninoculated controls (experiments 1 and 2) or in singly virus-inoculated pigs (experiment 3). In a first experiment with conventional pigs, the PRRSV-3d-H1N1 and PRRSV-7d-H1N1 infections induced mean body temperatures > 40.5 degrees C during 8 days (peaks 41.1 and 41.6 degrees C, respectively) and mean growth reductions of 3.4 and 4.8 kg, respectively, during the 2 weeks after H1N1, along with marked depression and respiratory disease. The PRRSV-14d-H1N1 infection, on the contrary, was largely subclinical. In a second experiment with conventional pigs, PRRSV-3d-H1N1 and PRRSV-7d-H1N1 infections were clinically milder, with smaller increases in mean body temperatures (peak 40.5 degrees C in both groups) and growth reductions (1.4 and 1.6 kg, respectively). In both groups, only one pig showed prominent general and respiratory signs. In a final experiment with CDCD pigs, PRRSV-7d-H1N1 infection had minimal effects on mean clinical performances and growth and, except for one pig that was severely affected, differences with the single virus inoculations were negligible. Thus, both the time interval between infections and the sanitary status of pigs can affect the clinical outcome of dual PRRSV-H1N1 infections. However, factors so far unknown seem to cause large variations in the clinical response between individual pigs. PMID- 15129584 TI - Identification of the tetracycline resistance gene, tet(M), in Erysipelothrix rhusiopathiae. AB - This is the first report to demonstrate the presence of tet(M) in naturally occurring isolates of tetracycline-resistant Erysipelothrix rbusiopathiae, which causes swine erysipelas. The tet(M) gene was isolated from E. rhusiopathiae strain KY5-42. The nucleotide and the deduced amino acid sequence were 99% identical to the tet(M) gene from Enterococcus faecalis. The gene was necessary and sufficient for the expression of tetracycline resistance in Escherichia coli. The presence of the tet(M) gene in the 114 tetracycline-resistant E. rhusiopathiae isolates from diseased pigs was detected by the polymerase chain reaction assay. The specific amplified DNA fragment was obtained from all 114 tetracycline-resistant strains. It was suggested that the tet(M) gene was widely present in the field isolates of E. rhusiopathiae resistant to tetracycline. PMID- 15129585 TI - Molecular epidemiology of contagious bovine pleuropneumonia in Tanzania based on amplified fragment length polymorphism and pulsed-field gel electrophoresis analysis. AB - The genetic diversity of 60 field strains of Mycoplasma mycoides ssp. mycoides, small colony type (M. mycoides SC), comprising 56 isolates from cattle in Tanzania, one from Kenya, two from Botswana and one from Portugal, as well as the type (PG1T) and vaccine (T1-SR49) strains, was investigated. The strains were analyzed for variations in the EcoRI and Csp6I restriction sites in the genomic DNA using the amplified fragment length polymorphism (AFLP) technique, and variations in the BamHI restriction sites using pulsed-field gel electrophoresis (PFGE). Six AFLP types were detected among the analysed strains. The AFLP profiles of the type and vaccine strains were indistinguishable from each other. Indistinguishable AFLP profiles were found for 55 Tanzanian field strains, one of them isolated in 1990 and the other 54 isolated in 1998/1999, although one strain isolated in 1999 showed a different profile. Strains from different countries revealed different AFLP profiles. Six PFGE types were detected among the analysed strains, with all the 56 Tanzanian field strains displaying indistinguishable PFGE profiles. Strains from different countries revealed different PFGE profiles, and so did the type and vaccine strains. The strong genomic homogeneity among M. mycoides SC strains associated with outbreaks of contagious bovine pleuropneumonia in different regions of Tanzania suggests that the outbreaks of the disease in the 1990-99 period might have been caused by a single epidemic clone. Moreover, this study has demonstrated that AFLP and PFGE are potential tools for molecular epidemiological studies of M. mycoides SC infections. PMID- 15129586 TI - Paratuberculosis in red deer (Cervus elaphus): an immunohistochemical study. AB - In the present study, we compared the utility of immunohistochemistry with serological and histological results for the characterization of Mycobacterium avium ssp. paratuberculosis (M. paratuberculosis) in tissues of affected red deer. Bacterial isolation was considered the standard reference. Samples were taken from seven clinically affected animals with typical macroscopic lesions. The enzyme linked immunosorbent assay (ELISA) and the gel diffusion tests (GD) were used for serological determinations. Samples from intestine and mesenteric lymph nodes were processed for bacterial isolation and histology. M. paratuberculosis was isolated from all the animals. Histologically, lymph nodes displayed necrosis and mineralization at the cortical and medullar areas. Ziehl Neelsen stained bacteria were numerous inside macrophages and Langhans-type giant cells. Giant and epithelioid cells and lymphocytes were prominent at the ileal mucous membrane. The immunostaining of M. paratuberculosis was very clear inside epithelioid and giant cells. Image analysis was carried out to determine the immunostained area. There was total agreement among the methods employed. Immunohistochemistry can be very useful when the microorganism cannot be recovered from tissues or faeces. PMID- 15129587 TI - Trial of targeting therapy against malignant glioma using monoclonal antibody. AB - Although the conventional treatment of malignant gliomas including surgery, radiotherapy and systemic chemotherapy has advanced, their current prognosis remains poor. The reactivity of monoclonal antibodies (mAbs) with human tumor cells may allow precise localization and appropriate therapy. We have developed several mAbs against malignant gliomas, and have reported the results of the experimental studies aimed at their clinical application as targeting therapy. The initial results of employing murine mAb 425 which binds to specifically to the epidermal growth factor (EGF) receptor in glioma therapy have been excellent, but less satisfactory, positively due to the immunogenicity of the murine mAbs, and limitations of the efficacy of the unmodified antibodies. Therefore, we also investigated the accumulation and the tumor suppression effect of human mAb CLNIgG and CLNIgG-drug (DXR: doxorubicin) conjugates to the tumor. The human mAb CLNIgG, derived from human uterine cancer lymph node cells, was found to bind strongly to human malignant glioma cells. PMID- 15129588 TI - [Prevention of pathogenic bacterial infection by using phage lytic enzyimes]. PMID- 15129589 TI - The effect of noise on the health of children. AB - The effects of noise on health, especially that of children, were reviewed. (1) From the point of view of disturbance of daily living, subjective recognition of "noisiness" is an important issue in relation to the study of noise. Concerning the effects of airplane noise on school children, while no effects on the hearing level were detected, a significant increase in the complaint of "noisiness" was observed. (2) Exposure of pregnant women to airplane noise was found to be associated with a decrease in the body weight of newborn babies. Moreover, the height of 3-year-old boys and girls was found to be significantly decreased in association with increase in the environmental noise. (3) Noise levels that seemed to have some influence on the sleep of adults did not affect the sleep of children. (4) In a group of children living in noisy districts exhibiting poor academic performance, the academic performance seemed to become progressively worse as the school grade advanced. (5) No consensus has been arrived at in regard to headphone-induced hearing impairment. Researches and studies effective enough to influence policy decisions must be continually conducted in the future, with appropriate control for related factors. PMID- 15129590 TI - Implementation of integrated medical curriculum in Japanese medical schools. AB - Recently, various integrated medical curricula, which can be defined as courses with subject matter classified by organ systems rather than according to departments such as surgery and internal medicine, are beginning to be introduced to bedside-learning in Japan. For example, in such an integrated medical curriculum, lectures in the course on neurological diseases would be given by a team that would include neurosurgeons, neurologists, and pathologists. Using medical education on neurological diseases as an example of an integrated medical curriculum, we analyzed the factors related to the neurological disease course as an example of an integrated medical curriculum in the clinical medicine course at our school. We also compared our course with those of all private medical schools in Japan, using the syllabuses of these private medical schools for the comparison, and considered elements that measured interdisciplinary participation in presenting the curriculum. For an integrated medical curriculum to gain interdisciplinary acceptance, the curriculum should be constituted using all medical disciplines related to the specific organ involved in the disease process under study, including both basic medicine and clinical medicine. In addition, teachers should be informed of the rationale for such a curriculum to promote their participation and a textbook on the integrated medical curriculum is needed. A curriculum committee should play an important role in promoting this type of medical education. PMID- 15129591 TI - Subjective evaluation and overall satisfaction after tympanoplasty for chronic simple suppurative otitis media. AB - A questionnaire survey was conducted in 324 patients with chronic, simple, suppurative otitis media who had undergone tympanoplasty 6 months or more previously to investigate post-operative hearing, tinnitus, vertigo, occlusive feeling of the ear and otorrhea. In addition, the overall satisfaction with tympanoplasty was assessed by VAS value. Subjective hearing improvement was observed in 73.1% of the patients whose hearing was poor and in 50% of those whose hearing was good before the operation. The degree of satisfaction assessed by VAS value corresponded with the subjective hearing assessment. As to tinnitus, 66.2% of the patients became aware of the disappearance or alleviation of symptoms. In the case of patients who had tinnitus before the operation, the degree of awareness of tinnitus and the degree of satisfaction assessed by VAS value coincided. However, no changes in the VAS value were observed in those who did not have tinnitus before the operation. As for vertigo, 30.5% of the patients who had vertigo preoperatively became aware of the disappearance of the symptoms after the operation. The degree of satisfaction assessed by VAS value corresponded with the presence or absence, severity and frequency of vertigo. As to the fullness of the ear, alleviation of the symptoms was subjectively noted by 85.9% of the patients who had symptoms before the operation. The degree of satisfaction assessed by VAS value corresponded with the severity of the symptoms in those who had symptoms before the operation. As for otorrhea, the disappearance of the symptoms was subjectively noted by 85.5% of the patients who had otorrhea before the operation. The degree of satisfaction assessed by VAS value corresponded with the post-operative changes in otorrhea. Based on the above results, it was assumed that the patients placed greatest expectation on hearing improvement when they underwent tympanoplasty. VAS is considered a useful method to evaluate the degree of satisfaction of patients after surgery. PMID- 15129592 TI - A comparative study on the observation of spontaneous nystagmus with Frenzel glasses and an infrared CCD camera. AB - OBJECTIVES: To compare the usefulness of a CCD camera with infrared illumination (IR-CCD camera) over Frenzel glasses (F Glasses) for the observation of spontaneous nystagmus, the incidence and direction of nystagmus, and the frequency, amplitude and slow phase of spontaneous nystagmus. METHODS: One hundred vertiginous patients, fifty-three females and forty-seven males participated in this study. Before undergoing routine neurotological examination, their eye movements were recorded by electronystagmogram (ENG) in conjunction with observations of eye movements under F glasses and through an IR-CCD camera. The data was collected from patients who exhibited spontaneous nystagmus either under F glasses or the IR-CCD camera. RESULTS: Thirty-three patients showed spontaneous nystagmus under F glasses. On the other hand, under the IR-CCD camera, all patients examined exhibited spontaneous nystagmus. The frequency of nystagmus was not significantly different between these two systems. However, the amplitude and slow phase velocity exhibited significantly larger values under the IR-CCD camera in patients with spontaneous nystagmus both under the IR-CCD camera and F glasses. CONCLUSION: From these observations and evidence, the IR-CCD camera can be recommended as a more useful system and powerful tool for neurotological examination than F glasses. PMID- 15129593 TI - Postoperative respiratory complications of video-assisted thoracic surgery for lung cancer. AB - PURPOSE: We analyzed the risk factors predisposing patients to develop postoperative respiratory complications (PRCs) in VATS lobectomy and segmentectomy for lung cancer, retrospectively. METHODS: Both univariate and multivariate analyses of PRCs were performed in seventy-five patients who had undergone VATS lobectomy and segmentectomy for lung cancer from November 1994 to December 2000. RESULTS: Univariate analysis of the development of PRCs revealed that the significant risk factors were age, ppo%VC, ppo%FEV, ppoFEV, poor pulmonary function, and duration of surgery. Multivariate logistic regression test in regard to the development of PRCs revealed that duration of surgery was the most significant risk factor. On the basis of the receiver operator characteristic analysis, duration of surgery more than 297 min had a sensitivity of 70% and a specificity of 66% for the development of PRCs. CONCLUSIONS: The duration of surgery should be less than five hours not to lose advantages of VATS lobectomy and segmentectomy. Therefore, if the duration of surgery is more than five hours for any reasons, conversion to limited thoracotomy or muscle-sparing methods is recommended. PMID- 15129594 TI - Usefulness of posturography after epidural block. AB - After a nerve block, observation of the course of effects is necessary until discharge of patients. Particularly epidural block in the lumbar region markedly affects ambulation and postural stability. Although there are few methods of objective evaluation of the postural stability, safe and early discharge is desired. We evaluated the influences of epidural block with 5 ml of 1% lidocaine on equilibrium before as well as 30, 60 and 90 minutes after epidural block. Computerized posturography allows the objective evaluation and quantitative assessment of impairment of receptors and the central nervous system involved in the maintenance of postural stability by analyzing of results. Locus length per unit area with the eyes open and that with the eyes closed 30 minutes after epidural block (27.339 +/- 11.761 cm and 25.804 +/- 10.561 cm, respectively) were significantly (P = 0.0067 and 0.0175, respectively) higher than baseline values (19.528 +/- 8.240 cm and 19.496 +/- 7.450 cm, respectively). Sway area with the eyes open 30 minutes after epidural block (3.923 +/- 2.494 cm2) was significantly (P = 0.0190) larger than the baseline value (2.533 +/- 1.309 cm2). These results suggest that marked effect remain after epidural block even when standing appears to be stable, and the observation of the course of effects after epidural block is still necessary for the safety of patients. We considered that locus length per unit area is a useful parameter for the assessment of an early discharge. PMID- 15129595 TI - Oral clonidine premedication exacerbates hypotension following tourniquet deflation by inhibiting noradrenaline release. AB - Clonidine premedication prevents tourniquet pain and reduces sympathetic nerve activity. We evaluated hemodynamic changes and catecholamine release following tourniquet deflation during spinal anesthesia in patients who received oral clonidine premedication. The final analysis included 24 otherwise healthy patients undergoing lower-limb surgery randomly assigned to two groups: those receiving approximately 5 micrograms/kg of oral clonidine 1 hr before anesthesia (clonidine group, n = 12), and those receiving no premedication (control group, n = 12). After lumbar anesthesia, a tourniquet was applied for approximately 60 minutes to each patient. Electrocardiogram, arterial blood pressure, and consumption of butorphanol for tourniquet pain were monitored. Blood samples were obtained at different times to measure serum concentration of catecholamine. In the clonidine group, mean blood pressure decreased from 87 +/- 7 mmHg at baseline to 65 +/- 10 mmHg after tourniquet deflation (P < 0.05). This peak reduction of mean blood pressure in the clonidine group was significantly lower than in the control group. After receiving clonidine premedication, the plasma noradrenaline concentrations in the clonidine group were significantly lower than those in the control group. Noradrenaline concentration increased in the control group from 162.3 +/- 89.2 pg/mL before tourniquet deflation to 199.3 +/- 95.7 pg/mL afterward (P < 0.01), but there was no significant change in noradrenaline concentration after tourniquet deflation in the clonidine group. We conclude that oral clonidine premedication exacerbated the reduction in mean blood pressure following tourniquet deflation by inhibiting noradrenaline release. PMID- 15129596 TI - Establishment of modified retroviral vector targeting X-linked severe combined immunodeficiency. AB - Gene therapy targeting hematopoietic stem cells has been proposed as a potential therapy for numerous genetic disorders affecting hematopoiesis. Moloney murine leukemia retroviral vectors are now widely used for clinical gene transfer into hematopoietic progenitors and progeny. However, maintaining expression of therapeutic genes inserted via moloney murine leukemia virus (MoMLV)-based vectors has proven to be more difficult than previously expected. In this study, an MND-IL-2R vector containing IL-2Rc gamma cDNA to treat X-linked severe combined immunodeficiency (X-SCID) was constructed from an MND vector that was modified by substituting the myeloproliferative sarcoma virus (MPSV) enhancer for that of MoMLV, deleting the negative control region located in the long terminal repeat (LTR) as an enhancer, and replacing the primer binding site (PBS) of MoMLV with the PBS of the endogenous murine retrovirus dl587rev. This vector was transduced into human CD34 + progenitor cells with comparable efficiency to that of the MoMLV-based vector. The use of this newly created vector may be advantageous for gene therapy of X-SCID. PMID- 15129597 TI - Anterior cruciate ligament injuries among wakeboarders: a case report. AB - No previous cases of anterior cruciate ligament (ACL) injuries sustained during wake-boarding have been reported. We report on a case involving an ACL injury sustained during wakeboarding. A 27-year-old man sustained an injury while attempting a wakeboarding maneuver(a heel-side back roll, consisting of a jump and simultaneous roll toward the heel side). He failed to complete his roll before landing, striking the water with his right shoulder foremost, then plunging underwater. When his wakeboard struck the water, his left knee was sprained by the rotational force exerted by the board. The patient was diagnosed with an isolated ACL injury and underwent arthroscopic ACL reconstructive surgery. The board used in wakeboarding is wider and subject to greater water resistance than that used in water skiing. The feet of the wakeboarder are firmly attached by binding boots to a board, laterally with respect to the direction of motion, impeding easy separation of the board from the feet in the event of a fall. Thus, wakeboarding conditions would appear to put wakeboarders at particular risk for ACL injuries. These conditions need be assessed from a medical perspective in order to devise ways to minimize the risk of such injuries. PMID- 15129598 TI - Re-evaluation of secondary amenorrheic patients one year after initial diagnosis: a prospective study. AB - OBJECTIVES: The aim of the study was to re-evaluate women with secondary amenorrhea one year after the first visit evaluation. STUDY METHODS: One hundred and seventy-five women with secondary amenorrhea were evaluated on the first visit. Their ages ranged from 18 to 29. Secondary amenorrhea was defined by the absence of menses for more than 3 months after excluding pregnancy. Women who were attempting to conceive were excluded from the study. 1) One hundred and two women were anovulatory (2) 36 had hypogonadotropic hypogonadism, 3) 11 had hypergonadotropic hypogonadism, and 4) 21 had hyperprolactinemia, 5) and five fell into other categories. The one hundred and forty-nine women in categories 1) to 3) were followed up for one year after the first diagnosis was made. A monthly progestational agent or HRT (hormone replacement therapy) was given to women with anovulation or hypogonadism, respectively. RESULTS: Of the 149 women in categories 1) to 3), 100 could be evaluated one year after the first diagnosis. There were 31 women whose diagnosis was changed. Anovulation changed to hypogonadotropic hypogonadism in 11 women, oligomenorrhea in four, and normal ovulatory cycle in two. Hypogonadotropic hypogonadism changed to anovulation in nine women, and to normal ovulatory cycle in one. Hypergonadotropic hypogonadism changed to normal ovulatory cycle in two women. CONCLUSIONS: A significant finding is that approximately one third of the women initially diagnosed with secondary amenorrhea, upon re-evaluation within one year had their diagnosis changed. Therefore evaluation of amenorrhea at an appropriate time is critical for proper management. PMID- 15129599 TI - A case report: change in fetal heart rate pattern on spontaneous uterine rupture at 35 weeks gestation after laparoscopically assisted myomectomy. AB - A 31-year-old nulligravid woman who underwent laparoscopically assisted myomectomy 5 months before becoming pregnant suffered uterine rupture at 35 weeks gestation. A 50 g intramuscular myomatous node had been removed laparoscopically. Early signs of rupture included sudden onset of severe abdominal tenderness and frequent uterine contractions despite reassuring FHR tracing. Variable deceleration was observed as late as 7.5 hours after onset. Emergency cesarean section was performed due to increasing severity of tenderness, revealing complete uterine rupture at the fundus site without extrusion of the fetus or placenta. A male neonate (2,860 g) was delivered without asphyxia and an Apgar score of 8. Total volume of hemorrhage was approximately 50 ml. The ruptured uterine wall was repaired by suturing in 2 layers. The present case indicates that sudden onset of abdominal tenderness in pregnant women with a history of laparoscopic myomectomy may suggest uterine rupture even in the presence of reassuring FHR. This is a rare case, as non-reassuring FHR patterns generally appear in the late stages of uterine rupture. PMID- 15129600 TI - The effect of mandibular advancement device on pharyngeal airway dimension in patients with congestive heart failure treated for sleep apnoea. AB - Continues positive airway pressure (CPAP) is recommended for treatment of sleep apnoea (SA) in patients with congestive heart failure (CHF) but is not easily tolerated resulting in poor patient compliance. Mandibular advancement device (MAD) is designed to inhibit pharyngeal airway (PAW) obstruction and may be a valuable alternative. It has been proposed that MAD exerts its effect by increasing PAW dimensions. This has not, however, been clearly demonstrated. The aim of this study was to examine the effect of MAD on PAW dimensions and SA in patients with CHF. Seventeen CHF-patients with mild to moderate heart failure, aged 68 +/- 6 years, (mean +/- SD), range 54-75 years, with sleep apnoea-hypopnea index (AHI) > or = 10 were evaluated. PAW dimensions were studied with and without the MAD, using lateral radiographs in supine position. Nocturnal breathing patterns were studied using a portable polysomnographic device during a single night with and without MAD. A reduction of AHI > or = 30% (arbitrary level) for each individual was regarded as a successful treatment. Mean AHI was reduced from 25.1 +/- 9.4 to 14.7 +/- 9.7 (p = 0.003). The PAW increased in its inferior section in 13 patients (p = 0.0001). AHI decreased > or = 30% in 9 patients (p = 0.003) of whom 8 showed increased PAW dimensions. Reduction of AHI was not significantly related to increased PAW dimensions. In conclusion MAD increased PAW dimensions and reduced SA in patients with CHF. The results may indicate that MAD reduces SA by other mechanism than increasing PAW dimensions. PMID- 15129601 TI - Treatment effect on signs and symptoms of temporomandibular disorders--comparison between stabilisation splint and a new type of splint (NTI). A pilot study. AB - The aim of the investigation was to compare the effect on signs and symptoms of temporomandibular disorders (TMD) of two different interocclusal appliances. Thirty patients with signs and symptoms of TMD received either a stabilisation splint or a new kind of splint based on the concept of nociceptive trigeminal inhibition (NTI). A clinical examination was performed and subjective symptoms were registered before start of treatment and after 3 and 6 months. All participants were offered to change to the other type of splint at the 3-month follow-up in case of no improvement or impairment of their symptoms. One subject in each group was lost early during the investigation. The mean time for taking impressions, to make inter-occlusal recording and to adjust the stabilisation splints was 17 minutes. The mean time to fit and adjust the NTI splints was 27 minutes. At the 3-month follow-up, 4 patients that had received NTI splints accepted the offer to change to stabilisation splints due to no improvement or impairment of their symptoms. These treatments were judged as failures. No one in the stabilisation splint group utilised the offer to change treatment. At the 6 month follow-up, 7 of the remaining 10 subjects with NTI splints reported some (n = 1) or significant (n = 6) improvement, 2 reported no change and one reported impairment. All 14 who had been treated with a stabilisation splint reported some (n = 2) or significant (n = 12) improvement. For all variables registered, the results were in favour for the stabilisation splint. One subject treated with a NTI splint exhibited an impaired occlusion at the 6-month follow-up. PMID- 15129602 TI - An alternative method for the fabrication of customised abutments for single tooth replacements. A clinical follow-up after 18 months. AB - The present paper describes an alternative method (The Cresco ST-method) for the fabrication of customised abutments designated for implant supported single-tooth (ST) replacements. The method is based on the conventional lost wax technology. 29 ST-replacements/crowns fabricated according to the method were tested/evaluated clinically during an 18 months period. The test was focused on mechanical complications. All crowns were supported by Cresco solid screw implants. Only one mechanical complication (retention screw loosening) was reported during the observation period. Based on the results it was concluded that the Cresco ST-method for fabrication of customised abutments is accurate and reliable. The influence of the interlocking features between implant and abutment and the advantages with customised abutments were discussed. PMID- 15129603 TI - Symptoms and signs of temporomandibular disorders in individuals with chronic whiplash-associated disorders. AB - The purpose of this study was to compare the prevalence of temporomandibular disorders (TMD) between individuals with chronic whiplash-associated disorders (WAD) and a group of age- and sex-stratified patients attending a Public Dental Service (PDS) clinic. Fifty-four individuals diagnosed with chronic WAD that were referred to a rehabilitation centre constituted the WAD group. The control group consisted of 66 patients at a PDS clinic (C group). Both groups underwent a standardised examination of the masticatory system comprising a questionnaire and a clinical examination. Eighty-nine per cent of the individuals in the WAD group had severe symptoms of TMD according to Helkimo's anamnestic index of dysfunction (A1) compared with 18% in the C group (p < 0.001). The individuals in the WAD group had also more signs of TMD. The maximum mouth opening capacity was 48 mm in the WAD group and 54 mm in the C group (p < 0.001). In the WAD group 17% had a mouth opening capacity < 40 mm compared with 2% in the C group (p < 0.05). Pain on palpation of the jaw muscles and on lateral palpation of the temporomandibular joints was more common in the WAD group (p < 0.001). Pain on mandibular mobility was reported by 30% in the WAD group and by 3% in the C group (p < 0.001). In conclusion, the prevalence of TMD was higher among individuals with chronic WAD compared with an age- and sex-stratified cohort of patients in a general dental practice. The results indicate that trauma to the neck also affects temporomandibular function. PMID- 15129604 TI - Factors that influence the proclination or retroclination of the lower incisors in children with prolonged thumb-sucking habits. AB - In some children with a prolonged finger-sucking habit, the lower incisors are retroclined, however, proclination of the lower incisors has been frequently observed. The purpose of this study was to determine how such differences develop in the child with a digit-sucking habit. Children with a prolonged thumb-sucking habit and proclination of the lower incisors (Ili/ML > 97 degrees) were cephalometrically and clinically compared to persistent thumb-suckers with retroclined lower incisors (Ili/ML < 89 degrees). In the group with retroclined lower incisors the angle between the thumb and the lower incisors was significantly smaller and the thickness of the lower lip significantly thinner than in the group with proclined incisors. A higher frequency of early loss of deciduous molars was also observed in the group with retroclined incisors. The tightness of the lower lip, early loss of deciduous molars and the angle between the teeth and the thumb are factors which have been identified to influence the inclination of the lower incisors in persistent digit suckers. The majority of children with prolonged thumb-sucking have proclined lower incisors rather than retroclined lower incisors. PMID- 15129605 TI - The relationship between hypodontia in the second premolar region and heredity of cleft, lip and palate in children with isolated cleft palate. AB - The aim of the investigation was to study the relationship between hypodontia in the second premolar region and heredity of cleft lip and palate (CLP) in children with isolated cleft palate (CP). The following null-hypothesis was tested: The heredity for CLP will not give rise to higher prevalence of hypodontia in the second premolar region of the maxilla and the mandible in CP-patients. The material consisted of 188 children born with non-syndromic CP (77 boys and 111 girls). Surgical files initiated at birth and panoramic radiographs at ten years of age have been studied concerning heredity for CLP and hypodontia of second premolars. For all CP-children, irrespective of hereditary background the prevalence of second premolar hypodontia did not differ significantly. However, there was a tendency to higher prevalence of hypodontia of the lower second premolars. The null-hypothesis: The heredity for CLP will not give rise to higher prevalence of hypodontia in the second premolar region of the maxilla and the mandible in CP-patients was not rejected. PMID- 15129606 TI - An innovative solution for skin tears: a case study. PMID- 15129608 TI - In pursuit of healthy bladder function. PMID- 15129607 TI - A new old problem. PMID- 15129609 TI - Treating wound deficit using constant tension to approximate wound margins. PMID- 15129610 TI - HIPAA and wound care. PMID- 15129611 TI - A critical review of natural therapies in wound management. AB - The use of natural medicine by both the general population and the nursing profession is increasing. Also referred to as unconventional, alternative, and complementary, these therapies often are derived from natural sources and facilitate improvements in health and well being by supporting innate healing processes of the body. Although the application of natural therapies in wound management is still in its infancy, several--honey, larval therapy, aromatherapy, herbal medicine, homeopathy, nutrition, and mind-body-spirit techniques--are demonstrating potential benefit in the management of acute and chronic wounds. Existing evidence of safety and effectiveness is predominantly based on the results of in vivo studies; confirmation from well-designed clinical trials is deficient. Consequently, clinical research is needed to examine these therapies to address the escalating incidence of chronic wounds in a climate of increasing healthcare costs. PMID- 15129612 TI - Living with diabetic foot ulcers: a life of fear, restrictions, and pain. AB - Lower extremity ulcers are a common and challenging problem for people with diabetes and clinicians who provide their care. A qualitative study of seven patients with type 1 or type 2 diabetes who had leg and/or foot ulcers was conducted to enhance understanding of the patient's perspective of living with lower extremity ulcers and diabetes. Data were collected through in-depth interviews guided by the patients' descriptive priorities. Results indicated that patients experienced changes in their feet, pain and insomnia, fatigue and limited mobility, social isolation and loneliness, a restricted life, loss of control, and fear for the future. When treating a patient's leg or foot ulcer, clinicians need to consider patients' subjective feelings toward the various aspects of their life situations and the impact of their situation on their quality of life. Understanding the ramifications of lower extremity ulcers and diabetes on patients is important to the health professional's ability to provide support during the illness process. PMID- 15129613 TI - Evidence for opioid variability, Part 2: Psychosocial influences. AB - There is substantial evidence to show that certain biological and psychosocial factors affect opioid requirements after surgery. In fact, evidence suggests that individuals are much more likely to be different rather than similar in how they sense pain, react to it and respond to therapy. In an earlier report (Seminars in Perioperative Nursing 10:3-16, 2001), we examined research related to the biological differences that explain variability in postoperative opioid use and defined relevant terminology. Here, we discuss the evidence that links psychosocial experiences to postoperative analgesic outcomes and pain, which include psychological states of patients, cultural influences and attitudes, and beliefs and biases held by both patients and health professionals. Content will assist perioperative nurses to understand the characteristics of their patients and circumstances that place patients at risk for needing increased analgesia or experiencing poor pain control. As perioperative nurses strive to integrate research into practice, it will be important to examine the results of research studies and to determine the usefulness of this information in developing individualized plans for postoperative pain management. PMID- 15129614 TI - Bioterrorism: an overview. AB - How real is the threat of bioterrorism? Experts may disagree on the likelihood of use, but the possibility cannot be totally dismissed. Complacent ignorance of a low-probability, high-cost risk is dangerous and can result in devastating global consequences. This is a US government work. There are no restrictions on its use. PMID- 15129616 TI - Learning style theories: matching preceptors, learners, and teaching strategies in the perioperative setting. AB - Individuals have unique learning styles that develop in childhood and remain constant throughout adulthood. One approach to orienting and training staff and students is to match individuals with a preceptor with a similar learning style. Benefits of matching learning styles and using appropriate teaching strategies for each learning style include decreased anxiety and increased staff and student satisfaction. This is a US government work. There are no restrictions on its use. PMID- 15129621 TI - [Issue dedicated to Aleksandr Abramovich Krasnovskii (1913-1993)]. PMID- 15129622 TI - [Mechanism of charge separation and their stabilization in bacterial reaction centers]. AB - The nuclear wavepacket formed by 20-fs excitation on the P* potential energy surface in native and mutant (YM210W and YM210L) reaction centers of Rhodobacter (Rb.) sphaeroides and Chloroflexus (C.) aurantiacus RCs was found to be reversibly transferred to the P+BA- surface at 120, 380, and 640-fs delays (monitored by measurements of BA- absorption at 1020-1028 nm). The reaction centers of YM210W(L) mutant show the most simple pattern of fs oscillations with a period of 230 fs in stimulated emission from P* and in the product P+BA-. The mechanisms of the electron transfer pathway between P* and BA and of the stabilization of the state P+BA- in bacterial reaction centers are discussed. PMID- 15129623 TI - [Dynamics of excitation in the photosystem I of cyanobacteria: transfer in the antenna, capture by the reaction site, and dissipation]. AB - The structure of a complex of photosystem I (PSI) of cyanobacteria and the mechanisms of the functioning of the antenna and PSI reaction site were described. The complex of PSI in thylakoids of cyanobacteia is organized as a trimer whose antenna is enriched in long-wave chlorophylls. The energy absorbed by these chlorophyls migrates to P700, inducing its oxidation. Long-wave chlorophyls are also involved in the dissipation of excessive energy; both the cation radical of P700 and the triplet of P700 effectively quench the fluorescence of long-wave chlorophyll of PSI. The energy exchange between the antennas of monomers in the trimer of PSI stimulates the dissipation of electron excitation energy, protecting the complex against photodestruction. The kinetics of energy migration in the antenna and charge separation in the reaction site of PSI trimers was studied using subpicosecond spectroscopy. Long-wave chlorophylls of PSI do not substantially affect the energy migration in the heterogeneous antenna of PSI but slow down the capture of energy of P700. The separation of changes in the reaction site of PSI is the most rapid among the known reaction sites. PMID- 15129624 TI - [Electrogenic reactions in the photosystem I]. AB - The processes of electron transfer in cyanobacterial photosystem I (PS I) and photoelectric methods of the studies were reviewed. Particular emphasis was placed on structural and kinetic characteristics of the electron transport chain. The electrogenicity in PS I complex and its interaction with natural donors (plastocyanin, cytochrome c6), natural acceptors (ferredoxin, flavodoxin), and artificial acceptors and donors (methyl viologen and other redox dyes) were studied. On the basis of photoelectric measurements and the X-ray structural data, the operating dielectric constants in the vicinity of charge carriers in situ were calculated. The profile of distribution of the dielectric constant along the PS I pigment-protein complex (from plastocyanin or cytochrome c6 through the chlorophyll dimer P700 to the acceptor complex) was estimated, and possible mechanisms of correlation between the local dielectric constant and the electron transfer rate constant in the corresponding segment of the chain were discussed. PMID- 15129625 TI - [Regulation of the primary photosynthesis processes]. AB - The mechanisms of primary processes of photosynthesis and macromolecular conformational changes that control the efficiency of primary energy transformation in photosynthesis are discussed. Special attention is focused on the analysis of chlorophyll fluorescence as an integrated parameter indicative of the efficiency and dynamics of primary steps of photosynthesis. Sharp changes in environmental conditions and other unfavorable factors may lead to the distortions of the coupling between consecutive electron transfer steps. As a result, an excess of electrons and/or electronic excitation energy may form at some sites of the electron transport chain. This may lead to the generation of reactive oxygen species responsible for the subsequent oxidative stress. The results of the application of these data in the areas of biotechnology and ecology are demonstrated. PMID- 15129626 TI - [Organization of the photosynthetic units based on action spectra of the functional activity]. AB - The methods for the analysis of the light-harvesting properties of the photosynthetic apparatus by measuring the optical cross-sections and the spectra of action of specific photoreactions were described. A short review summarizes our own findings obtained by these methods in the study of the organization of photosynthetic units in whole cells of purple bacteria, cyanobacteria, and green algae, and in isolated thylakoids from higher plants. PMID- 15129627 TI - [Visual transduction and calcium ions]. AB - A general scheme of visual induction and its regulation in mammalian retinal rods and the main signal proteins involved in the functioning of the visual signaling apparatus were considered. The role of calcium ions and calcium-binding proteins in the switching off of the visual signal and the transition of the photoreceptor cell from a photoexcited to a "dark" state was discussed in detail. PMID- 15129628 TI - [Phototaxis of the green algae: the new class of rhodopsin receptors]. AB - Photomotility behavior in green flagellate algae is mediated by rhodopsin-like receptors, which was initially suggested on the basis of physiological evidence. The cascade of rapid Ca(2+)-dependent electrical responses in the plasma membrane plays a key role in the signal transduction chain during both phototaxis and the photophobic response. The photoreceptor current through the plasma membrane is the earliest detectable event upon photoexcitation of the photoreceptors. Analysis of this current revealed that it consists of at least two components with different characteristics. Genes encoding two archaeal-type rhodopsins (type I rhodopsins) were recently identified in the genome of Chlamydomonas reinhardtii and named (Chlamydomonas Sensory Rhodopsins A and B CSRA and CSRB). The measurements of photoelectric and motor responses in genetic transformants of C. reinhardtii enriched in each of these receptor proteins showed that the two components of the photoreceptor current are mediated by the two rhodopsins, and that both CSRA and CSRB are involved in phototaxis and the photophobic response. The CSRA-mediated current dominates at high light intensities and contributes primarily to the photophobic response. The CSRB-initiated transduction involves an efficient amplification cascade and mediates the highly sensitive phototaxis at low light intensities. CSRA and CSRB expressed heterologously in oocytes of Xenopus laevis act as light-gated proton channels, although it is unclear whether this channel activity plays a functional role in the initiation of motor responses and/or occurs in the native system. PMID- 15129629 TI - [Phytochrome regulation of the plant phospholipase activity]. AB - The interaction of external stimuli with receptors of plant cell surface can activate the enzymes of lipid metabolism such as phospholipases C and D. The products of the catalysis, i.e., posphoinositide metabolites, phosphatidic acid, fatty acids, etc. participate in signal transduction as secondary messengers related to numerous regulatory processes. One of the physiologically important factors of regulatory control in plants is light, which plays the crucial role in triggering the cellular signaling network. This review presents the information on phospholipid signaling in plants, which is connected with light transduction processes occurring with the involvement of the plant regulatory pigment phytochrome. PMID- 15129630 TI - [Photodynamic activity and singlet oxygen]. AB - The primary mechanisms for the photodynamic action of pigments and dyes, the principles of their division into mechanisms of type I and type II, and the role of these processes in biological systems are reviewed. Singlet oxygen is considered to be an indicator of the mechanisms of photodynamic reactions. The methods of its detection are described, which are based on the use of chemical traps, measurements of infrared phosphorescance at 1270 nm, and the registration singlet oxygen-sensitized delayed fluorescence caused by the summation of the energy of two singlet oxygen molecules by one dye molecule. PMID- 15129631 TI - [Photobiophysics of furanocoumarins]. AB - Furocoumarins (psoralens) are photosensitizers of plant origin, which increase the sensitivity of biological objects to near ultraviolet (UV-A, 320-400 nm). In combination with UV-A, they are successfully used for treating many dermal and autoimmune diseases (PUVA therapy and photophoresis). Along with therapeutic effects, the furocoumarin photochemotherapy induces a number of side-effects (erythema, edema, hyperpigmentation, and premature aging of skin). All photobiological effects of furocoumarins result from their photochemical reactions. Therefore, in order to advance the therapy, it is necessary to know the photochemical mechanisms of induction of both side- and therapeutic effects. The types of photoreactions of furocoumarins classified with respect to reactive photoproducts interacting with substrate were considered. Primary emphasis was placed on reactions proceeding with the participation of photooxidation products of furocoumarins. Among these photoproducts, at least two types can be distinguished. Some of them possess membranotoxic properties, others produce the immunosuppressory action in vivo. The photochemical mechanisms of the formation of the photoproducts of furocoumarins are different. It was found that, by varying the illumination conditions (intensity of UV-A radiation or the concentration of the photosensitizer), it is possible to obtain the photoproducts of furocoumarins that have either membranotoxic or immunosuppressory properties. It was found that the mechanisms of the immunosuppressive action of the photooxidation products of furocoumarins have some features in common with those underlying the PUVA therapy and photophoresis. It is assumed that the photochemical basis of the therapeutic action of furocoumarins is the reactions with the involvement of the products of their photooxidation. PMID- 15129632 TI - [Molecular and cellular mechanisms of the low intensity laser radiation effect]. AB - The main aspects of the free radical conception of the molecular and cellular mechanisms of the stimulating action of low-intensity radiation in the red region of the spectrum were considered. These are: (1) Primary acceptors of incident radiation are endogenous porphyrins, which may act as photosensitizers giving initiator-radicals for secondary free radical reactions. (2) Target cells for light irradiation during quantum therapy may be blood leukocytes, fibroblasts, keratinocytes, endotheliocytes, etc. (3) The initiation of the secondary free radical reactions due to lipid peroxidation of cell membranes (in particular, of leukocytes) brings about an increase in ion permeability including that for calcium. The increase in intracellular calcium concentration leads to phagocytes priming, i.e., to increased production of reactive oxygen species (ROS) under subsequent stimulation of the cell. (4) Photosensitized generation of ROS in the cytoplasm of some cells induces a free-radical activation of synthesis of proteins, the most significant in the light of the present concept being the de novo synthesis of inducible NO-synthase, superoxide dismutase, and various cytokines. The experimental evidence for the basic statements of the conception of free radical mechanisms for the stimulating action of low-intensity laser and noncoherent radiations is presented. A relation between the primary mechanisms of the stimulating action of light and the secondary effects that determine the sanative effect of quantum therapy in the process of wound healing (bactericidity, cell proliferation, and improved microcirculation) was established. Moreover, it was shown that nitrosyl complexes of heme proteins, such as hemoglobin and cytochrome c, are the primary chromophores of laser radiation. Upon irradiation, they can easily dissociate to produce free nitric oxide. In turn, released nitric oxide may be responsible for blood vessel relaxation and activation of mitochondrial respiration. This phenomenon is just observed during phototherapy by means of low-intensity laser radiation. PMID- 15129633 TI - [Approaches to the targeted intracellular delivery of photosensitizers in order to enhance their efficacy and cell specificity]. AB - The main physicochemical properties of photosensitizers used in the photodynamic therapy of cancer and their subcellular distribution after in vitro and in vivo administration were analyzed. It was shown that the effect of photosensitizers is realized at very short distances from the sites of their intracellular localization, and the sensitivities of different cellular compartments to the photocytotoxic action of photosensitizers are different. The necessity of intranuclear delivery of photosensitizers into the nuclei of target cells in order to enhance their efficacy and cell specificity was shown and the available approaches to the targeted delivery of photosensitizers were analyzed. The mechanisms of nucleocytoplasmic transport through the nuclear pore complex, which can be used for the delivery of photosensitizers inward the nucleus, are reviewed. Different modular transporters for photosensitizers comprising (i) a ligand module, which binds to an internalizable receptor overexpressed on the target cells, (ii) an intracellular localization signal, (iii) a carrier module, and (iv) an endosomolytic module were characterized. All these modules were shown to be fully functional within the chimeric polypeptide and the polypeptide as a whole. A significant enhancement of photocytotoxicity and cell specificity of photosensitizers delivered by these transporters were demonstrated. The transporters described represent a new generation of pharmaceuticals which can be widely used for targeted drug delivery. PMID- 15129634 TI - [Current trends in photodynamic therapy of neoplasms and non-neoplastic diseases]. AB - The photodynamic therapy of tumors is a modern therapeutic modality for organ preserving treatment of oncological diseases. The method is based on selective laser irradiation of tumor tissues previously sensitized by tumorotropic dyes. During the last decades, photodynamic therapy has become worldwide known as a proper approach to the treatment of the patients with malignant tumors of various locations and a number of nontumoral diseases. The characteristics of modern photosensitizers and light sources for photodynamic therapy and their clinical applications are reviewed. PMID- 15129635 TI - Process power. PMID- 15129636 TI - Clinical management. Where medicine meets management. Let us play. AB - There is evidence that hospital play hastens children's recovery, increases compliance and reduces the need for general anaesthesia. The children's national service framework recommends that all children in hospital have daily access to a play specialist. A survey showed an 11 per cent rise in the number of play specialists in the past two years, but more are needed. PMID- 15129637 TI - DTC commissioning. An arranged marriage. AB - The first privately run diagnostic and treatment centre required nine months of complex talks between the strategic health authority, the trust and Bupa. The contract cost more than if it had been run by the trust, but it increased access and transferred risk. One lesson was that staff should have been better acclimatised to the idea. PMID- 15129638 TI - Mental health. Right on whose side. PMID- 15129639 TI - HSJ people. Stretcher cases. PMID- 15129640 TI - My brilliant career--finance. In it for the long run. Interview by Emma Forrest. PMID- 15129642 TI - Finance. Rural pocket. PMID- 15129641 TI - Finance. Everything in its place. PMID- 15129643 TI - Finance. Stars and gripes. PMID- 15129644 TI - Finance. Code breakers. PMID- 15129645 TI - Possibility that transmissible viruses might be involved in the etiology of human cancer. PMID- 15129646 TI - A journey with T cells, primate/human retroviruses and other persisting human T cell tropic viruses. AB - A study of the growth of primate/human T cells led to mechanisms for temporary laboratory culture of these cells (discovery of interleukin-2) and also their continuous culture (by immortalization after infection with human T-cell lymphotropic virus type 1 or 2 (HTLV-1 or 2)). Cultures of lymphocytes also led us to isolate five persisting T-tropic viruses: 1. the Hall's Island strain of gibbon ape leukemia virus, 2. HTLV-1, 3. HTLV-2, 4. human immunodeficiency virus and 5. human herpes virus-6 (HHV-6). This report is a brief synopsis of the discoveries of the first human retroviruses, the HTLV. PMID- 15129647 TI - HTLV-1 and associated adult T-cell leukemia/lymphoma. AB - Human T-cell leukemia/lymphoma virus type 1 (HTLV-1) infects 15-20 millions individuals worldwide. This oncoretrovirus can be transmitted through 3 ways: horizontally, vertically (mother to child) and via blood transfusion. HTLV-1 causes 2 major diseases: adult T-cell leukemia/lymphoma (ATLL) and tropical spastic paraparesis/HTLV-1-associated myelopathy. Tax is a 40-kDa phosphoprotein that is encoded by the pX region of the virus. Several lines of evidence have demonstrated a central role for this protein in the immortalization or transformation of the HTLV-1 infected cells. Apart from its ability to drive transcription from the viral promoter, it also deregulates the cell cycle, inhibits apoptosis, has an effect on the maintenance of the genomic stability and induces the production of several cytokines. In addition, several arguments strongly suggest the existence of host genetic factors, that could be involved in the HTLV-1 infection as well as in the development of ATLL among HTLV-1 infected individuals. ATLL can be classified into 4 major subtypes: a smoldering type, a chronic type, a lymphoma type and a leukemic type. The demonstration by Southern blot analysis of the clonal integration of an HTLV-1 provirus in the tumoral cells represents the gold-standard to define biologically ATLL. The survival rate of ATLL patients, especially those who develop the acute leukemic or lymphomas forms, is very poor, and such clonal malignant CD4 expansion remains one of the most severe lymphoproliferations. PMID- 15129648 TI - Epstein-Barr virus-associated lymphoproliferative disorders. AB - Epstein-Barr virus (EBV) is a ubiquitous member of the herpesvirus family that is associated with a variety of lymphomas and lymphoproliferative diseases. It encodes a multitude of genes that drive proliferation or confer resistance to cell death. Among these are two key viral proteins which mimic the effects of the activated cellular signaling proteins. EBV-associated lymphomas include Burkitt's lymphoma; natural killer (NK)/T-cell lymphoma, lymphoma and lymphoproliferative diseases in immunocompromized populations, and Hodgkin's lymphoma. The character of the viral association differs among these entities with some consistently associated with EBV in all populations and all parts of the world, and others associated with the virus only in particular circumstances. An example of the former is nasal NK/T-cell lymphoma, while an example of the latter is Burkitt's lymphoma. The pattern of viral gene expression also varies among tumor types with different viral genes playing key roles in different tumors and conferring sensitivity to immune surveillance. Thus some of the post-transplant lymphoproliferative diseases are exquisitely sensitive to CD8 T-cell immunosurveillance, while other tumors such as Burkitt's lymphoma may be nearly impervious to such surveillance. Knowledge of the EBV association is not only important for understanding the pathogenesis of these tumors, but is increasingly important for diagnosis, monitoring and treatment. PMID- 15129649 TI - Human herpesvirus 8 (HHV-8/KSHV) and hematologic malignancies. AB - Human herpesvirus 8 (HHV-8), also defined Kaposi's sarcoma (KS)-associated herpesvirus, was identified by Chang and colleagues in 1994 using purely molecular techniques, before any serological evidence or virus isolation in cell culture could be achieved. HHV-8 is unique among herpesviruses because its prevalence in the general population is low and because it possesses the richest weaponry of viral oncogenes and tumor-promoting factors ever described. Eleven HHV-8-specific genes are homologs of cellular genes, which were hijacked from the host during a long parallel evolution, and at least five of such genes show both in vitro and in vivo transforming ability. HHV-8 is the causative agent of KS, but it has also been associated with different hematologic malignancies, including primary effusion lymphoma (PEL), multicentric Castelman's disease (MCD), MCD-related immunoblastic/plasmablastic lymphoma and various atypical lymphoproliferative disorders. Although low-level silent infection was detected in bone marrow stromal cells from patients with multiple myeloma, a role of HHV-8 in this disease is unlikely. As seen with KS, the incidence of HHV-8-associated lymphoproliferative disorders is increased in the setting of human immunodeficiency virus infection. PMID- 15129650 TI - HCV-associated lymphomas. AB - Epidemiologic and molecular observations have recently suggested that hepatitis C virus (HCV) may be the causative agent of some B-cell non-Hodgkin lymphomas (B NHL). Epidemiologic data suggest that in Italy about 5% of B-NHL are caused by HCV. Molecular data indicate a close relationship between HCV-associated B-NHL and type II mixed cryoglobulinemia. The latter disorder appears to reflect the benign monoclonal proliferation of B cells expressing a specific cross-reactive idiotype, known as WA, that may recognize an antigen of HCV, perhaps the E2 protein. Genetic abnormalities occurring during this phase of antigen-induced clonal expansion may drive the neoplastic transformation into low- or high-grade lymphoma. The recent demonstration that splenic B cell lymphomas associated with HCV-infection may regress after successful antiviral therapy confirms a role for this virus in B-cell lymphomagenesis. PMID- 15129651 TI - [Guideline for conducting diagnostic and therapeutic examinations in hemodynamics]. PMID- 15129652 TI - Eur(omega)conference Angiogenesis II. PMID- 15129653 TI - [Against the routine induction of labor at 41 weeks]. PMID- 15129654 TI - [Recommendations for clinical practice: management of extra-uterine pregnancy]. PMID- 15129655 TI - Cohort effect in 10 years mortality in elderly people. PMID- 15129656 TI - [Scientia rehabilis. 5. German Ophthalmology Academy]. PMID- 15129661 TI - [Proper management of self-pay patients]. PMID- 15129662 TI - [More time for physicians via ambulatory health centers]. PMID- 15129663 TI - Midwives take action against a major cause of maternal death: a new partnership campaign to prevent postpartum haemorrhage. PMID- 15129664 TI - Improving access to information for midwives in developing countries. PMID- 15129665 TI - Measuring the outcome of pregnancy in Europe. PMID- 15129666 TI - Abstracts of the 70th Annual meeting of the German Society of Cardiology--Heart and Circulatory Research. 15-17 April 2004, Mannheim, Germany. PMID- 15129667 TI - Ethics of caesarean section in the USA. PMID- 15129669 TI - Abstracts of the Canadian Ophthalmological Society Annual Meeting. Vancouver, Canada, 16-19 June 2004. PMID- 15129668 TI - Abstracts of the British Maternal and Fetal Medicine Society 9th Annual Conference. Glasgow, United Kingdom, 1-2 April 2004. PMID- 15129670 TI - Halcyon days. PMID- 15129671 TI - Emergency airway access equipment. PMID- 15129673 TI - Modeling health insurance expansions: effects of alternate approaches. AB - Estimates of the costs and consequences of many types of public policy proposals play an important role in the development and adoption of particular policy programs. Estimates of the same, or similar, policies that employ different modeling approaches can yield widely divergent results. Such divergence often undermines effective policymaking. These problems are particularly prominent for health insurance expansion programs. Concern focuses on predictions of the numbers of individuals who will be insured and the costs of the proposals. Several different simulation-modeling approaches are used to predict these effects, making the predictions difficult to compare. This paper categorizes and describes the different approaches used; explains the conceptual and theoretical relationships between the methods; demonstrates empirically an example of the (quite restrictive) conditions under which all approaches can yield quantitatively identical predictions; and empirically demonstrates conditions under which the approaches diverge and the quantitative extent of that divergence. All modeling approaches implicitly make assumptions about functional form that impose restrictions on unobservable heterogeneity. Those assumptions can dramatically affect the quantitative predictions made. PMID- 15129674 TI - Federal aid strengthens health care safety net: the strong get stronger. AB - Two new federal initiatives--community health center expansion and Community Access Program grants--have improved access to care for low-income people and strengthened linkages among safety net providers, according to findings from the Center for Studying Health System Change's (HSC) 2002-03 site visits to 12 nationally representative communities. Grant recipients have added services to fill safety net gaps or to improve collaboration among safety net providers. However, communities with weaker safety nets were less likely to receive federal aid, and funding for both programs is limited, hampering the potential impact on the nation's system of care for low-income and uninsured people. PMID- 15129675 TI - Preferred provider organizations and Medicare: is there an advantage? AB - A key component of the new Medicare reform law is an overhaul of Medicare managed care, including a strong emphasis on recruiting private plans--especially preferred provider organizations (PPOs)--to participate in the new Medicare Advantage program. Citing the popularity of PPOs for privately insured Americans, proponents have touted PPOs as critical to injecting more and better competition into Medicare. This study, based on findings from the Center for Studying Health System Change's (HSC) site visits to 12 nationally representative communities, explores the reasons for the strong growth in commercial PPO enrollment and examines whether PPOs--as currently structured--can add value to Medicare. The available evidence suggests that the PPO model will face challenges in achieving the policy goals set forth in the Medicare Prescription Drug, Improvement and Modernization Act of 2003 (MMA), including increasing benefits, improving quality and slowing cost growth. PMID- 15129672 TI - DNA vaccine encoding human immunodeficiency virus-1 Gag, targeted to the major histocompatibility complex II compartment by lysosomal-associated membrane protein, elicits enhanced long-term memory response. AB - Antigen presentation by major histocompatibility complex type II (MHC II) molecules and activation of CD4+ helper T cells are critical for the generation of immunological memory. We previously described a DNA vaccine encoding human immunodeficiency virus-1 p55Gag as a chimera with the lysosome-associated membrane protein (LAMP/gag). The LAMP/gag chimera protein traffics to the MHC II compartment of transfected cells and elicits enhanced immune responses as compared to a DNA vaccine encoding native gag not targeted to the MHC II compartment. We have now investigated the long-term responses of immunized mice and show that the LAMP/gag DNA vaccine promotes long-lasting B cell- and CD4+ and CD8+ T-cell memory responses induced by DNA encoding non-targeted Gag decay rapidly and elicit very low or undetectable levels of gag DNA is sufficient to generate T-cell memory. Following this initial priming immunization with LAMP/gag DNA, booster immunizations with native gag DNA or the LAMP/gag chimera are equally efficient in eliciting B- and T-cell secondary responses, results in accordance with observations that secondary expansion of CD8+ cells in the boost phase does not require additional CD4+ help. These findings underscore the significance of targeting DNA-encoded vaccine antigens to the MHC II processing compartments for induction of long-term immunological memory. PMID- 15129676 TI - A phase I, dose escalation trial of ZD0473, a novel platinum analogue, in combination with gemcitabine. AB - PURPOSE: To develop a combination regimen for clinical testing, we performed a dose escalation study of ZD0473 in combination with gemcitabine. ZD0473 is a novel platinum analogue with an aliphatic cyclic carrier ligand. In vitro and in vivo studies suggest that it possesses a different spectrum of antitumor activity from cisplatin and carboplatin. In single-agent studies of ZD0473, myelosuppression was the predominant toxicity and responses wer observed. METHODS: In this combination phase I trial, 36 patients with advanced cancer were accrued to four dose levels, with doses of ZD0473 and gemcitabine ranging from 60 to 120 mg/m2 and 600 to 750 mg/m2, respectively ZD0473 was administered on day 1 and gemcitabine was given on days 1 and 8 of a 21-day cycle. RESULTS: Hematologic toxicity was dose-limiting. Grade 3 and 4 thrombocytopenia and neutropenia occurred during 60% and 41% of all cycles. Nonhematologic toxicities were mild and reversible. Two partial responses and 19 patients with stable disease were observed. CONCLUSIONS: The recommended phase II doses are 90 mg/m2 of ZD0473 and 750 mg/m2 of gemcitabine for lightly pretreated patients and 600 mg/m2 for heavily pretreated patients. The combination of ZD0473 and gemcitabine is associated with dose-dependent thrombocytopenia and neutropenia as well as having promising clinical activity. PMID- 15129677 TI - Abstracts of the 44th Austrian Geriatric Congress with International Participation. Bad Hofgastein, 20-24 March 2004. PMID- 15129678 TI - Abstracts of the 3rd EORTC-NCI International Meeting on Cancer Molecular Markers. April 18-20, 2004, Brussels, Belgium. PMID- 15129680 TI - Abstracts from the American Telemedicine Association 9th Annual Meeting. May 2-5, 2004, Tampa, Florida, USA. PMID- 15129679 TI - Selected abstracts on myeloma and leukemia from the American Society of Hematology annual meeting, 2003. PMID- 15129681 TI - Abstracts of the 52nd annual meeting and postgraduate course of the Pacific Coast Reproductive Society. April 28-May 2, 2004. Rancho Mirage, California, USA. PMID- 15129682 TI - Recommendations for better asthma care and control made by working groups at the Caribbean Asthma and Allergy Workshop. PMID- 15129683 TI - [Abstracts of the 46th National Congress on Anesthesia and Resuscitation. Paris, France, 17-18 April 2004]. PMID- 15129684 TI - Abstracts of the 39th Annual Meeting of the European Association for the Study of the Liver. April 14-18, 2004, Berlin, Germany. PMID- 15129685 TI - ABRF 2004. Integrating Technologies in Proteomics and Genomics. February 28-March 2, 2004, Portland, Oregon, USA. Abstracts. PMID- 15129686 TI - Abstracts of the 12th Annual Conference of the Indian Association for the Study of the Liver. Lucknow, March 12-14, 2004. PMID- 15129687 TI - A perspective on the current state of death education. AB - The author offers some views on the current state of death education with focus on the sparing attention given the death education of health professionals and of grief counselors. There is need for improved integration of the knowledge accumulated in the study of death, dying, and bereavement into the basic curricula of the parent disciplines and professional schools. Facilitation of personal engagement with the issue of mortality is an important component of the educative process. Various assessment problems are outlined and some suggestions for improvements are offered. The death education needs of various groups, including school age children and older adults, are noted. The article contains a list of references, many not cited in the text, recommended for an extensive review of developments in death education. PMID- 15129688 TI - Psychological research on death attitudes: an overview and evaluation. AB - One of the most substantial legacies of Herman Feifel was his pioneering research on attitudes toward death and dying in a variety of populations. The authors review the large and multifaceted literature on death anxiety, fear, threat and acceptance, focusing on the attitudes toward death and dying of relevant professional and patient groups, and the relationship of death concern to aging, physical and mental health, religiosity, and terror management strategies. We conclude with several recommendations for improving the conceptual and practical yield of future work in this area. PMID- 15129689 TI - Meanings of death seen through the lens of grieving. AB - Through the lens of grief we can discern many meanings of death, human existence, suffering, the life of the deceased, the life of the mourner, and love. This essay summarizes what the author, in part inspired by Herman Feifel, has learned about such meanings in nearly three decades of thinking, teaching, and writing about grieving as an active response to what happens in bereavement and the suffering that loss entails. It recasts his rethinking of grief within the broad categories of philosophical reflection, including meta-theory and philosophy of science, conceptual analysis, existential-phenomenological analysis, philosophy of mind, epistemology, metaphysics, and ethics. PMID- 15129690 TI - Recovery following bereavement: an examination of the concept. AB - Herman Feifel noted that appropriate attention to one's mourning and grieving allowed the dead to die and the bereaved "to redefine and reintegrate oneself into life" (H. Feifel, 1977, p. 9). The author takes this central focus on bereavement outcomes as the springboard for an examination of the concept recovery following bereavement. He examines the meanings of the terms recover and bereavement and considers the centrality of concepts from life span human development, the life crisis literature, and existential phenomenology for defining the full possibilities of the concept of recovery following bereavement. Seminal ideas from Alexander Leighton and Thomas Attig are examined for their power to provide operational definitions for Feifel's idea about redefining and reintegrating oneself into life as the full meaning of the concept recovery following bereavement. PMID- 15129691 TI - Death writ large. AB - Mainstream thanatology has devoted its efforts to improving the understanding, care, and social integration of people who are confronted with life-threatening illness or bereavement. This article suggests that it might now be time to expand the scope and mission to include large-scale death and death that occurs through complex and multi-domain processes. Obstacles to developing a systematic macrothanatology are identified. The 9-11-01 terrorist attacks on America are discussed as an example of mass death with complex correlates and consequences. Other examples are taken from the realms of war, disease, disaster, and extinction. PMID- 15129692 TI - Analysis. Suspended sentence. PMID- 15129693 TI - [Diagnoses and surgery of chronic sinusitis in children]. PMID- 15129694 TI - [The basic clinical research of head-neck neoplasms]. PMID- 15129695 TI - Molecular imaging in oncology by means of nuclear medicine: fact or fiction? PMID- 15129696 TI - Usefulness of brain 99mTc-TRODAT-1 SPET for the evaluation of Parkinson's disease. AB - Nigral dopaminergic projections to the striatum are targeted in Parkinson's disease (PD). The extent of the degeneration of the dopaminergic system in PD can be visualised by dopamine transporter imaging using single-photon emission tomography (SPET). In this study in 188 patients with PD, we analysed the image patterns and compared them with the clinical features in order to verify the usefulness of technetium-99m TRODAT-1 brain SPET in the evaluation of patients with PD. Two independent readers visually assessed SPET slices from three brain axes according to a "fine" visual scale; results were also grouped according to a "rough" visual scale. Results of both visual and semi-quantitative analyses were compared among patients with different stages of PD and healthy controls. There was good agreement between the readers in the interpretation of the image patterns [kappa statistic (kappa)=0.85 for the presence of PD; kappa=0.88 for the rough scale and 0.81 for the fine scale]. Good concordance was obtained when visual interpretation was used to evaluate the presence of PD (sensitivity = 98%, specificity = 86%, kappa = 0.85). Semi-quantitative analyses revealed significant negative correlations between both striatal and putaminal uptake and disease severity as assessed using the Hoehn and Yahr scale (rho = -0.89 and -0.93 respectively). An apparent decrease in striatal uptake in early PD, hardly discernible from the uptake level in advanced PD, was commonly found in visual analyses. The results suggest that both visual and semi-quantitative analyses of 99mTc-TRODAT-1 SPET images reflect neurodegeneration in PD, and that 99mTc-TRODAT 1 SPET represents an adequate means for evaluation of the status of patients with PD. PMID- 15129697 TI - Heterogeneity of cerebral blood flow in frontotemporal lobar degeneration and Alzheimer's disease. AB - This study was designed to quantify the heterogeneity on cerebral blood flow single-photon emission tomography (SPET) images in frontotemporal lobar degeneration (FTLD) and Alzheimer's disease (AD) using a three-dimensional fractal analysis. Twenty-one FTLD patients, 21 AD patients and 11 healthy controls underwent technetium-99m hexamethylpropylene amine oxime SPET scanning. Patients with FTLD and AD matched for sex, age and the severity of dementia as estimated with the Clinical Dementia Rating and were determined to be in the early stage of illness. We delineated the SPET images using a 35% cut-off and a 50% cut-off of the maximal voxel radioactivity and measured the number of voxels included in the contours of two different cut-offs. The fractal dimension (FD) was calculated by relating the logarithms of the cut-offs and the numbers of voxels, and it was defined as the heterogeneity of the cerebral perfusion. We divided the SPET images into two sets, anterior and posterior, with equal numbers of coronal SPET slices. We calculated total FD, anterior FD and posterior FD for total, anterior and posterior SPET images. Anterior FDs for FTLD and AD were 1.55 +/- 0.34 and 1.24 +/- 0.19 (P = 0.0002). The ratios of anterior to posterior FD for FTLD and AD were 1.81 +/- 0.41 and 1.32 +/- 0.14 (P < 0.0001). Use of the anterior FD and the ratio of anterior to posterior FD separated FTLD patients from AD patients and controls with a sensitivity of 85.7% and a specificity of 93.8%. Anterior FD and the ratio of anterior to posterior FD may be useful in distinguishing FTLD from AD. PMID- 15129698 TI - Comparison of methodologies for the in vivo assessment of 18FLT utilisation in colorectal cancer. AB - Fluorine-18 3'-deoxy-3'-fluorothymidine (18FLT) is a tissue proliferation marker which has been suggested as a new tumour-specific imaging tracer in positron emission tomography (PET). The objectives of this study were to investigate the pharmacokinetics of 18FLT in patients with colorectal cancer, defining methodologies for the quantitative analysis of the in vivo 18FLT uptake and subsequently assessing the accuracy of semi-quantitative measures. Dynamic acquisitions over a single field of view of interest identified by computed tomography were carried out for up to 60 min following injection of 18FLT (360 +/ 25 MBq). Dynamic arterial blood sampling was carried out in order to provide a blood input function. Simultaneous venous samples were also taken in order to investigate their potential utilisation in deriving a hybrid input function. Arterial and venous blood samples at 5, 15, 30, 60 and 90 min p.i. were used for metabolite analysis. Eleven patients with primary and/or metastatic colorectal cancer were studied on a lesion by lesion basis (n = 21). All acquired images were reconstructed using ordered subsets expectation maximisation and segmented attenuation correction. Time-activity curves were derived by image region of interest (ROI) analysis and image-based input functions were obtained using abdominal or thoracic aorta ROIs. Standardised uptake values (SUVs) were calculated to provide semi-quantitative indices of uptake, while non-linear regression (NLR) methodology in association with a three-compartment model and Patlak analysis were carried out to derive the net influx constant Ki. The metabolite analysis revealed two radioactive metabolites, with the parent compound representing approximately 80% of the total radioactivity in the 30-min plasma sample. In the case of NLR, better fits were obtained with a 3k model (i.e. k4 = 0) for both lesion and bone marrow time-activity curves. For the same lesions, a high correlation was observed between the Ki derived from either Patlak analysis or NLR(3k) and the corresponding SUVs. Our results also suggest that the quantitative behaviour of 18FLT in vivo (up to 60 min p.i.) may be characterised using a 3k model or Patlak analysis in combination with image derived input functions. The good correlation found between the SUVs (at 60 min) and Ki values supports the use of semi-quantitative indices to assess the proliferation rate of colorectal cancer lesions in vivo with 18FLT. PMID- 15129699 TI - [18F]FDG in recurrent breast cancer: diagnostic performances, clinical impact and relevance of induced changes in management. AB - Prognosis and management of patients with recurrent breast cancer depend on the spread of the disease. The aim of this study was to evaluate the performance of fluorine-18 fluorodeoxyglucose gamma camera positron emission tomography (FDG GPET) in detecting breast cancer recurrence, its clinical impact and the relevance of induced changes in management. Patients (n = 134) with suspicion of recurrence either clinically or on conventional imaging (suspected recurrence: SR) or with an isolated increase in tumour marker levels (occult recurrence: OR) underwent FDG-GPET on a coincidence gamma camera. The reference standard for evaluation of accuracy, either histology (n = 26) or follow-up for 1 year (n = 49), was available in 75 (56%) patients. A questionnaire was sent to the referring clinician to evaluate the impact of FDG on management. Responses were obtained for 75 patients. Information regarding both approaches was available for 46 patients (46/134 = 34%). At the patient level, the sensitivity of FDG-GPET was 84%, significantly higher than the 63% sensitivity for conventional modalities, and the specificity was 78% versus 61%. The values for FDG-GPET were 81% and 86% respectively in the SR group and 90% and 73% respectively in the OR group, without any significant difference between these settings. The rate of change in management was 44% overall, 43% in the SR group and 45% in the OR group. Within the two groups, intermodality (major) changes were more frequent than intramodality (minor) changes. In the 46 patients for whom both approaches were available, 93% of management modifications were relevant (validated by biopsy or clinical follow-up). The results of this retrospective study show that FDG-GPET has an important role to play in patient management by confirming and evaluating the extent of recurrence or by localising occult recurrence. PMID- 15129700 TI - FDG PET imaging for grading and prediction of outcome in chondrosarcoma patients. AB - The aims of this study were to assess the potential of fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) for tumor grading in chondrosarcoma patients and to evaluate the role of standardized uptake value (SUV) as a parameter for prediction of patient outcome. FDG PET imaging was performed in 31 patients with chondrosarcoma prior to therapy. SUV was calculated for each tumor and correlated to tumor grade and size, and to patient outcome in terms of local relapse or metastatic disease with a mean follow-up period of 48 months. Chondrosarcomas were detectable in all patients. Tumor SUV was 3.38 +/- 1.61 for grade I (n = 15), 5.44 +/- 3.06 for grade II (n = 13), and 7.10 +/- 2.61 for grade III (n =3). Significant differences were found between patients with and without disease progression: SUV was 6.42 +/- 2.70 (n = 10) in patients developing recurrent or metastatic disease compared with 3.74 +/- 2.22 in patients without relapse (P = 0.015). Using a cut-off of 4 for SUV, sensitivity, specificity, and positive and negative predictive values for a relapse were 90%, 76%, 64%, and 94%, respectively. Combining tumor grade and SUV, these parameters improved to 90%, 95%, 90%, and 95%, respectively. Pretherapeutic tumor SUV obtained by FDG PET imaging was a useful parameter for tumor grading and prediction of outcome in chondrosarcoma patients. The combination of SUV and histopathologic tumor grade further improved prediction of outcome substantially, allowing identification of patients at high risk for local relapse or metastatic disease. PMID- 15129701 TI - [18F]FDG PET as a substitute for second-look laparotomy in patients with advanced ovarian carcinoma. AB - The aim of this study was to compare the prognostic value of fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) with that of second look laparotomy (SLL) in patients with advanced ovarian carcinoma following primary chemotherapy. Fifty-five patients who had undergone cytoreductive surgery and adjuvant chemotherapy for advanced ovarian carcinoma were enrolled in the study. Thirty patients underwent SLL after primary treatment (SLL group), while 25 underwent FDG PET after primary treatment without SLL (PET group) We retrospectively reviewed the medical records of the 55 patients for comparison of progression-free interval and disease-free interval between the two groups. Ovarian carcinomas recurred in 37 of the 55 patients. When the progression-free interval and the disease-free interval in patients in the PET group were compared with those in the SLL group, no significant differences were observed. The progression-free interval in the PET and SLL groups were 28.8 +/- 12.7 months and 30.6 +/- 13.7 months, respectively (P = 0.29). The disease-free interval in the negative PET group was 40.5 +/- 11.6 months, and that in the negative SLL group was 48.6 +/- 12.1 months (P = 0.12). In conclusion, FDG PET has a similar prognostic value to SLL, and can substitute for SLL in the follow-up of patients who have had ovarian carcinoma, especially when there is a high risk for recurrence. PMID- 15129702 TI - Differentiation of mediastinal FDG uptake observed in patients with non-thoracic tumours. AB - In regions with a high prevalence of granulomatous diseases, benign inflammatory fluorine-18 fluorodeoxyglucose (FDG) uptake in the mediastinum is frequently observed even in healthy subjects. We examined parameters of mediastinal FDG uptake to determine whether they can differentiate malignancy from benign lesions. Seventy patients with non-thoracic tumours who had mediastinal uptake on FDG positron emission tomography (PET) were included (33 males, 37 females; age 57.5 +/- 16.9 years; 168 lymph nodes). Determination of metastasis was confirmed by biopsy or computed tomography (CT) follow-up over 12 months (metastasis, 29; benign lesions, 41). No significant difference between the metastasis group and the benign group was found in terms of residual disease in the primary site (48% vs 46%), lung invasion (29% vs 20%), number of sites of uptake (2.3 vs 2.4), smoking history (30.3% vs 46.3%) or bilateral uptake (52% vs 54%). Maximal standardised uptake values (SUVs) in the mediastinal metastasis group were higher (4.9 +/- 1.8) than those in the benign group (2.5 +/- 0.9) (P < 0.05). Using 3.4 as a cut-off value for maximal SUV, a sensitivity of 86% and a specificity of 85% were achieved (AUC = 0.917). Maximal SUV showed better predictive value than lymph node size measured on chest CT (P < 0.05). In 8 of 51 normal subjects who underwent FDG PET as a routine check-up, mediastinal FDG uptake was observed. Maximal SUV in normal subjects was 2.5 +/- 0.8, which was similar to that in the benign group. In conclusion, maximal SUV was identified as a significant parameter for determining whether mediastinal FDG uptake represents malignant metastasis. When maximal SUV exceeded 3.4, the metastasis rate was high regardless of lymph node size. PMID- 15129704 TI - A realistic 3-D gated cardiac phantom for quality control of gated myocardial perfusion SPET: the Amsterdam gated (AGATE) cardiac phantom. AB - A realistic 3-D gated cardiac phantom with known left ventricular (LV) volumes and ejection fractions (EFs) was produced to evaluate quantitative measurements obtained from gated myocardial single-photon emission tomography (SPET). The 3-D gated cardiac phantom was designed and constructed to fit into the Data Spectrum anthropomorphic torso phantom. Flexible silicone membranes form the inner and outer walls of the simulated left ventricle. Simulated LV volumes can be varied within the range 45-200 ml. The LV volume curve has a smooth and realistic clinical shape that is produced by a specially shaped cam connected to a piston. A fixed 70-ml stroke volume is applied for EF measurements. An ECG signal is produced at maximum LV filling by a controller unit connected to the pump. This gated cardiac phantom will be referred to as the Amsterdam 3-D gated cardiac phantom, or, in short, the AGATE cardiac phantom. SPET data were acquired with a triple-head SPET system. Data were reconstructed using filtered back-projection following pre-filtering and further processed with the Quantitative Gated SPECT (QGS) software to determine LV volume and EF values. Ungated studies were performed to measure LV volumes ranging from 45 ml to 200 ml. The QGS-determined LV volumes were systematically underestimated. For different LV combinations, the stroke volumes measured were consistent at 60-61 ml for 8-frame studies and 63-65 ml for 16-frame studies. QGS-determined EF values were slightly overestimated between 1.25% EF units for 8-frame studies and 3.25% EF units for 16-frame studies. In conclusion, the AGATE cardiac phantom offers possibilities for quality control, testing and validation of the whole gated cardiac SPET sequence, and testing of different acquisition and processing parameters and software. PMID- 15129703 TI - Requirements for clinical PET: comparisons within Europe. AB - The aim was to assess the requirements for a positron emission tomography (PET) cancer imaging service. The UK was used as an example to create a mathematical model for calculating the number of dedicated PET scanners and cyclotron/radiochemistry production facilities required to support the demand for PET studies in lung cancer. This was then extended to all oncological indications for PET and comparison was made with present infrastructure in the UK and Europe. A clinical algorithm for the use of PET in lung cancer management was created and built into a comprehensive computer model with variable parameters. From lung cancer incidences, data reported in the literature and local data, the proportion of patients following each algorithmic path was determined and used to calculate the number of PET scans and hence PET scanners required for lung cancer, and all cancer indications. Substituting lung cancer incidences, the PET infrastructure required for each European country was assessed. From this analysis, 29,886 PET scans per year for lung cancer investigation (provision of 12 scanners) and 121,589 PET scans (2,026.5 per million population) for all indications [provision of 49 scanners (0.82 per million population)] are required in the UK; at present there are seven scanners, and thus 42 new scanners are required. Results reported here demonstrate considerable lack of investment in PET in Europe, with marked variation; Belgium has the most sufficient infrastructure (197.80% of requirements), and excluding France, which is soon to see extensive development, the UK has the least sufficient infrastructure (14.39% of requirements). Considerable investment is required so that cancer management can gain the clinical and cost-effective benefit of this functional imaging technique, which has been established. PMID- 15129705 TI - Quantification of myocardial perfusion defects using three different software packages. AB - Software packages are widely used for quantification of myocardial perfusion defects. The quantification is used to assist the physician in his/her interpretation of the study. The purpose of this study was to compare the quantification of reversible perfusion defects by three different commercially available software packages. We included 50 consecutive patients who underwent myocardial perfusion single-photon emission tomography (SPET) with a 2-day technetium-99m tetrofosmin protocol. Two experienced technologists processed the studies using the following three software packages: Cedars Quantitative Perfusion SPECT, Emory Cardiac Toolbox and 4D-MSPECT. The same sets of short axis slices were used as input to all three software packages. Myocardial uptake was scored in 20 segments for both the rest and the stress studies. The summed difference score (SDS) was calculated for each patient and the SDS values were classified into: normal (< 4), mildly abnormal (4-8), moderately abnormal (9-13), and severely abnormal (> 13). All three software packages were in agreement that 21 patients had a normal SDS, four patients had a mildly abnormal SDS and one patient had a severely abnormal SDS. In the remaining 24 patients (48%) there was disagreement between the software packages regarding SDS classification. A difference in classification of more than one step between the highest and lowest scores, for example from normal to moderately abnormal or from mildly to severely abnormal, was found in six of these 24 patients. Widely used software packages commonly differ in their quantification of myocardial perfusion defects. The interpreting physician should be aware of these differences when using scoring systems. PMID- 15129706 TI - 99mTc-ciprofloxacin planar and tomographic imaging for the diagnosis of infection in the postoperative spine: experience in 48 patients. AB - The non-invasive assessment of postoperative spinal infections can pose a substantial diagnostic challenge, especially in the presence of orthopaedic devices. Whereas white blood cell scanning is of limited use in the spine, the low normal bone marrow uptake of technetium-99m ciprofloxacin combined with its claimed bacterial specificity makes it theoretically an ideal candidate for the evaluation of postoperative spinal infections. This study aimed to evaluate 99mTc ciprofloxacin planar and single-photon emission tomography (SPET) imaging in relation to microbiological and clinical diagnosis in the postoperative spine. Planar imaging was performed at 1, 3 and 24 h and SPET was performed at 3 h post injection of 370 MBq 99mTc-ciprofloxacin. Images were scored by two independent certified nuclear medicine physicians, blinded to the final diagnosis. Within the 48 patients, there were 13 deep infections. Sensitivity, specificity and accuracy at visual scoring were respectively 54%, 71% and 67% (1 h), 62%, 77% and 73% (3 h), 42%, 91% and 77% (24 h) for planar imaging and 100%, 74% and 81% for SPET. When recently operated patients (< 6 months) were excluded, the specificity of the SPET imaging rose to 81%. In conclusion, unlike white blood cell scanning, 99mTc-ciprofloxacin SPET is sensitive in evaluating infections in the postoperative spine. Sensitivity is much higher for SPET than for planar imaging. However, the results presented prove that its specificity is limited, especially in recently operated patients. Taking this limitation into account, we advise planar and SPET imaging at 3 h post injection and an interval of at least 6 months after surgery to minimise the likelihood of false positives. PMID- 15129707 TI - Assessment of regional lung functional impairment with co-registered respiratory gated ventilation/perfusion SPET-CT images: initial experiences. AB - In this study, respiratory-gated ventilation and perfusion single-photon emission tomography (SPET) were used to define regional functional impairment and to obtain reliable co-registration with computed tomography (CT) images in various lung diseases. Using a triple-headed SPET unit and a physiological synchroniser, gated perfusion SPET was performed in a total of 78 patients with different pulmonary diseases, including metastatic nodules (n = 15); in 34 of these patients, it was performed in combination with gated technetium-99m Technegas SPET. Projection data were acquired using 60 stops over 120 degrees for each detector. Gated end-inspiration and ungated images were reconstructed from 1/8 data centered at peak inspiration for each regular respiratory cycle and full respiratory cycle data, respectively. Gated images were registered with tidal inspiration CT images using automated three-dimensional (3D) registration software. Registration mismatch was assessed by measuring 3D distance of the centroid of the nine selected round perfusion-defective nodules. Gated SPET images were completed within 29 min, and increased the number of visible ventilation and perfusion defects by 9.7% and 17.2%, respectively, as compared with ungated images; furthermore, lesion-to-normal lung contrast was significantly higher on gated SPET images. In the nine round perfusion-defective nodules, gated images yielded a significantly better SPET-CT match compared with ungated images (4.9 +/- 3.1 mm vs 19.0 +/- 9.1 mm, P<0.001). The co-registered SPET-CT images allowed accurate perception of the location and extent of each ventilation/perfusion defect on the underlying CT anatomy, and characterised the pathophysiology of the various diseases. By reducing respiratory motion effects and enhancing perfusion/ventilation defect clarity, gated SPET can provide reliable co-registered images with CT images to accurately characterise regional functional impairment in various lung diseases. PMID- 15129708 TI - Preliminary results of transarterial rhenium-188 HDD lipiodol in the treatment of inoperable primary hepatocellular carcinoma. AB - A multicentre study was sponsored by the International Atomic Energy Agency (Vienna) to assess the safety and efficacy of trans-arterial rhenium-188 HDD conjugated lipiodol (radioconjugate) in the treatment of patients with inoperable hepatocellular carcinoma (HCC). The radioconjugate was prepared by using an HDD (4-hexadecyl 1-2,9,9-tetramethyl-4,7-diaza-1,10-decanethiol) kit developed in Korea, and lipiodol. Over a period of 18 months, 70 patients received at least one treatment of radioconjugate. Some patients were re-treated if there was no evidence of disease progression. The level of radioconjugate administered was based on radiation-absorbed dose to critical normal organs, calculated following a "scout" dose of radioconjugate. The organs at greatest risk for radiation toxicity are the normal liver, the lung and the bone marrow. An Excel spreadsheet was used to determine maximum tolerated activity (MTA), defined as the amount of radioactivity calculated to deliver no more than 12 Gy to lungs, or 30 Gy to liver, or 1.5 Gy to bone marrow. These doses have been found to be safe in multiple trials using external beam therapy, but this has not been confirmed for systemically administered radiopharmaceuticals. Patients were followed for at least 12 weeks after therapy, until recovery from all toxicity. The clinical parameters evaluated included toxicity, response as determined by contrast enhanced computed tomography, palliation of symptoms, overall survival, performance status (Karnofsky) and hepatic function (Child's classification). Liver function tests, serum alpha-fetoprotein (AFP) levels and complete blood counts were done at each follow-up visit. In the majority of patients, the scout dose studies indicated the radiation absorbed dose to normal liver to be the limiting factor to the treatment dose, while in a few patients dose to lung was the limiting factor. Radiation dose to bone marrow was negligible and was thus not a factor for the MTA calculations. Side-effects were minimal and usually presented as loss of appetite, right hypochondrial discomfort and low-grade fever, even at high levels of administered radioactivity. The symptoms resolved with simple supportive therapy within 3 days of onset. Liver function tests at 24 and 72 h showed no significant changes and complete blood counts at 1 week, 4 weeks and 12 weeks showed no changes (no bone marrow suppression). Sixteen patients were treated in the dose escalation phase of the study, when the activities administered started at 1.8 GBq (50 mCi) and rose to 7.7 GBq (206 mCi). In the efficacy phase of the study a further 54 patients were treated. Both groups of patients are included in this paper. The treatment activity of 188Re lipiodol administered transarterially ranged from 1.8 to 9.8 GBq (50-265 mCi), with a mean activity of 4.6 GBq (124 mCi). Survival at 3 months was 90%, and at 6 months, 60%; 19% survived for 1 year. Mean survival after treatment in the total treated group of 70 patients was 9.5 months, with a range of 1-18 months. The results of this multicentre study show that 188Re-lipiodol is a safe and cost effective method to treat primary HCC via the transarterial route. In terms of efficacy, it is potentially a new therapeutic approach for further evaluation by treatment of larger numbers of patients. PMID- 15129709 TI - Neurofunctional imaging of the pancreas utilizing the cholinergic PET radioligand [18F]4-fluorobenzyltrozamicol. AB - The pancreas is one of the most heavily innervated peripheral organs in the body. Parasympathetic and sympathetic neurons terminate in the pancreas and provide tight control of endocrine and exocrine functions. The aim of this study was to determine whether the pancreas can be imaged with a radioligand that binds to specific neuroreceptors. Using fluorine-18 4-fluorobenzyltrozamicol (FBT), which binds to the presynaptic vesicular acetylcholine transporter, positron emission tomography scans were performed in four adult mice, two adult rhesus monkeys, and one adult human. In these mammals, the pancreas is intensely FBT avid, with uptake greater than in any other organ at 30, 60, and 90 min. The maximum standardized uptake value (SUV) ratios of pancreas to liver, for example, ranged from 1.4 to 1.7 in rhesus monkeys (mean 1.6; median 1.7) and from 1.9 to 4.7 (mean 3.24; median 3.02) in mice. The maximum SUV ratio of pancreas to liver in the human was 1.8. These data suggest that neuroreceptor imaging of the pancreas in vivo is feasible in animal models and humans. This imaging could allow researchers to interrogate functions under control of the autonomic nervous system in the pancreas, with applications possible in transplanted and native pancreata. Also, as beta cell function is intimately related to parasympathetic cholinergic input, FBT activity in the pancreas may correlate with insulin producing beta cell mass. This could ultimately provide a method of in vivo imaging in animal models and humans for diabetes research. PMID- 15129711 TI - Usefulness of lymphoscintigraphy and intraoperative gamma probe detection in the identification of sentinel nodes in cervical cancer. PMID- 15129712 TI - Health technology assessment of PET in oncology: re Eur J Nucl Med Mol Imaging 2003; 30:637-641. PMID- 15129713 TI - Undergraduate teaching of nuclear medicine in the Czech Republic. PMID- 15129710 TI - Myocardial perfusion scintigraphy: the evidence. AB - This review summarises the evidence for the role of myocardial perfusion scintigraphy (MPS) in patients with known or suspected coronary artery disease. It is the product of a consensus conference organised by the British Cardiac Society, the British Nuclear Cardiology Society and the British Nuclear Medicine Society and is endorsed by the Royal College of Physicians of London and the Royal College of Radiologists. It was used to inform the UK National Institute of Clinical Excellence in their appraisal of MPS in patients with chest pain and myocardial infarction. MPS is a well-established, non-invasive imaging technique with a large body of evidence to support its effectiveness in the diagnosis and management of angina and myocardial infarction. It is more accurate than the exercise ECG in detecting myocardial ischaemia and it is the single most powerful technique for predicting future coronary events. The high diagnostic accuracy of MPS allows reliable risk stratification and guides the selection of patients for further interventions, such as revascularisation. This in turn allows more appropriate utilisation of resources, with the potential for both improved clinical outcomes and greater cost-effectiveness. Evidence from modelling and observational studies supports the enhanced cost-effectiveness associated with MPS use. In patients presenting with stable or acute chest pain, strategies of investigation involving MPS are more cost-effective than those not using the technique. MPS also has particular advantages over alternative techniques in the management of a number of patient subgroups, including women, the elderly and those with diabetes, and its use will have a favourable impact on cost effectiveness in these groups. MPS is already an integral part of many clinical guidelines for the investigation and management of angina and myocardial infarction. However, the technique is underutilised in the UK, as judged by the inappropriately long waiting times and by comparison with the numbers of revascularisations and coronary angiograms performed. Furthermore, MPS activity levels in this country fall far short of those in comparable European countries, with about half as many scans being undertaken per year. Currently, the number of MPS studies performed annually in the UK is 1,200/million population/year. We estimate the real need to be 4,000/million/year. The current average waiting time is 20 weeks and we recommend that clinically appropriate upper limits of waiting time are 6 weeks for routine studies and 1 week for urgent studies. PMID- 15129714 TI - Detection of alterations in brain glucose metabolism by positron emission tomography in Takayasu's arteritis. PMID- 15129715 TI - 99mTc-EDDA/HYNIC-TOC scintigraphy in oncological diagnostics: methodological considerations. PMID- 15129716 TI - Re: controversies--[Tc(CO)3]+ chemistry: a promising new concept for SPET? PMID- 15129717 TI - Effect of increased 99mTc/99Tc ratios on count rates in sentinel node procedures: a randomised study. PMID- 15129718 TI - Precise localisation of osteoblastoma with SPET/CT. PMID- 15129719 TI - Construction and validation of cDNA-based Mt6k-RIT macro- and microarrays to explore root endosymbioses in the model legume Medicago truncatula. AB - To construct macro- and microarray tools suitable for expression profiling in root endosymbioses of the model legume Medicago truncatula, we PCR-amplified a total of 6048 cDNA probes representing genes expressed in uninfected roots, mycorrhizal roots and young root nodules [Nucleic Acids Res. 30 (2002) 5579]. Including additional probes for either tissue-specific or constitutively expressed control genes, 5651 successfully amplified gene-specific probes were used to grid macro- and to spot microarrays designated Mt6k-RIT (M. truncatula 6k root interaction transcriptome). Subsequent to a technical validation of microarray printing, we performed two pilot expression profiling experiments using Cy-labeled targets from Sinorhizobium meliloti-induced root nodules and Glomus intraradices-colonized arbuscular mycorrhizal roots. These targets detected marker genes for nodule and arbuscular mycorrhiza development, amongst them different nodule-specific leghemoglobin and nodulin genes as well as a mycorrhiza-specific phosphate transporter gene. In addition, we identified several dozens of genes that have so far not been reported to be differentially expressed in nodules or arbuscular mycorrhiza thus demonstrating that Mt6k-RIT arrays serve as useful tools for an identification of genes relevant for legume root endosymbioses. A comprehensive profiling of such candidate genes will be very helpful to the development of breeding strategies and for the improvement of cultivation management targeted at increasing legume use in sustainable agricultural systems. PMID- 15129720 TI - Evaluation of the GFP signal and its aptitude for novel on-line monitoring strategies of recombinant fermentation processes. AB - A high number of economically important recombinant proteins are produced in Escherichia coli based host/vector systems. The major obstacle for improving current processes is a lack of appropriate on-line in situ methods for the monitoring of metabolic burden and critical state variables. Here, a pre evaluation of the reporter green fluorescent protein (GFP) was undertaken to assess its use as a reporter of stress associated promoter regulation. The investigation of GFP and its blue fluorescent variant BFP was done in model fermentations using E. coli HMS 174(DE3)/pET11 aGFPmut3.1 and E. coli HMS174(DE3)/pET1aBFP host/vector systems cultured in fed-batch and chemostat regime. Our results prove the suitability of the fluorescent reporter proteins for the design of new strategies of on-line bioprocess monitoring. GFPmut3.1 variant can be detected after a short lag-phase of only 10 min, it shows a high fluorescence yield in relation to the amount of reporter protein, a good signal to noise ratio and a low detection limit. The fluorescence-signal and the amount of fluorescent protein, determined by ELISA, showed a close correlation in all fermentations performed. A combination of reporter technology with state of the art sensors helps to develop new strategies for efficient on-line monitoring needed for industrial process optimisation. The development of efficient monitoring will contribute to advanced control of recombinant protein production and accelerate the development of optimised production processes. PMID- 15129721 TI - Detection of specific DNA sequences using dual-color two-photon fluorescence correlation spectroscopy. AB - Fluorescence correlation spectroscopy (FCS) is rapidly growing in popularity as a biomedical research tool. FCS measurements can produce an accurate characterization of the chemical, physical, and kinetic properties of a biological system. They can also serve as a diagnostic, detecting particular molecular species with high sensitivity and specificity. We here demonstrate that dual-color FCS measurements can be applied to detect and quantify the concentration of specific non-fluorescent molecular species without requiring any modifications to the molecule of interest. We demonstrate this capability by applying dual-color two-photon fluorescence cross-correlation spectroscopy to detect single stranded gamma tubulin DNA in solution with high sensitivity. This quantification is independent of molecular size, and the methods introduced can be extended to measurements in complex environments such as within living cells. PMID- 15129722 TI - A de novo designed N-terminal disulphide bridge stabilizes the Trichoderma reesei endo-1,4-beta-xylanase II. AB - We have successfully engineered a disulphide bridge into the N-terminal region of Trichoderma reesei endo-1,4-beta-xylanase II (XYNII) by substituting Thr-2 and Thr-28 with cysteine. The T2C:T28C mutational changes increased the half-life in thermal inactivation of this mesophilic enzyme from approximately 40 s to approximately 20 min at 65 degrees C, and from less than 10 s to approximately 6 min at 70 degrees C. Therefore, the N-terminal disulphide bridge enables the use of XYNII at substantially higher temperatures than permitted by its native mesophilic counterpart. Altogether, thermostability increased by about 15 degrees C. The kinetic properties of the mutant XYNII were maintained at the level of the wild type enzyme. Our findings demonstrated that a properly designed disulphide bridge, here within the N-terminal region of XYNII, can be very effective in resisting thermal inactivation. PMID- 15129723 TI - Enhanced electron-transfer reactivity of horseradish peroxidase in phosphatidylcholine films and its catalysis to nitric oxide. AB - Direct electrochemistry of horseradish peroxidase (HRP) embedded in film of phosphatidylcholine (PC) is investigated at a pyrolytic graphite electrode by voltammetric methods. The electron-transfer reactivity between incorporated HRP and the electrode is found to be greatly enhanced by phosphatidylcholine film. Cyclic voltammetry (CV) of this incorporated peroxidase shows a pair of well defined and nearly reversible peaks, and the cathodic and anodic peak potentials are located at about -0.261 and -0.180 V, respectively versus saturated calomel electrode at pH 5.5. Ultraviolet-visible absorption spectra indicate that the heme microenvironment of HRP in phosphatidylcholine film is similar to that of its native status. It is also observed that HRP modified electrode is able to catalyze the electrochemical reduction of nitric oxide. Experimental results reveal that the peak current related to nitric oxide reduction is linearly proportional to its concentration in the ranges of 2.0 x 10(-7) -5.0 x 10(-6) mol (-1) and 2.0 x 10(-5) -1.0 x 10(-4) mol(-1), based on which an unmediated biosensor for nitric oxide is developed. PMID- 15129724 TI - Rapid and sensitive detection of 17beta-estradiol in environmental water using automated immunoassay system with bacterial magnetic particles. AB - A fully automated immunoassay of 17beta-estradiol (E2) was performed using anti E2 monoclonal antibody immobilized on bacterial magnetic particles (AntiE2-BMPs) and alkaline phosphatase-conjugated E2 (ALP-E2). E2 concentration in environmental water samples was evaluated by decrease in luminescence based on competitive reaction. A linear correlation between the luminescence intensity and E2 concentration was obtained between 0.5 and 5 ppb. The minimum detectable concentration of E2 was 20 ppt. All measurement steps were done within 0.5 h. The analysis of environmental water samples by a commercially available ELISA kit and the BMP-based immunoassay gave good correlation plots with a correlation efficient of 0.992. These results suggest that the fully automated system using the BMP-based immunoassay has some advantages in the high rapidity and sensitivity of the measurement. This system will enable us to determine low E2 concentrations without sample condensation. PMID- 15129725 TI - Improved stability of aerobic granules by selecting slow-growing nitrifying bacteria. AB - This study investigated the feasibility of improving the stability of aerobic granules through selecting slow-growing nitrifying bacteria. For this purpose, four sequencing batch reactors were operated at different substrate N/COD ratios ranging from 5/100 to 30/100. Results showed that aerobic granules formed in all four reactors, and aerobic granulation was a gradual process evolving from the dispersed seed sludge to mature and stable granules, and the whole granulation process could be divided into three phases, i.e. acclimation phase, granulation followed by granule maturation. The observed growth rate and mean size of mature aerobic granules were found to decrease as the substrate N/COD ratio was increased, while nitrifying population was enriched markedly in aerobic granules developed at high substrate N/COD ratios. The enriched nitrifying population in aerobic granules was responsible for the observed low growth rate of aerobic granules. It seems certain that the substrate N/COD ratio is an important factor in selecting nitrifying bacteria in aerobic granules. Aerobic granules with low growth rates showed strong structure and good settleability in terms of specific gravity, SVI and cell hydrophobicity that further lead to high stability as compared to those having high growth rates. This study demonstrated that the selection of slow-growing nitrifying bacteria through controlling substrate N/COD ratio would be a useful strategy for improving the stability of aerobic granules. PMID- 15129726 TI - Rational elimination of Aspergillus terreus sulochrin production. AB - Elimination of undesirable co-metabolites from industrial fermentations is often required due to the toxicities associated with the contaminants and/or due to difficulties in removing the contaminants during downstream processing. Sulochrin is a co-metabolite produced during the Aspergillus terreus lovastatin fermentation. Examination of the sulochrin biosynthetic pathway identifies the emodin anthrone polyketide synthase (PKS) at the origin. Thus, genetically disrupting the emodin anthrone PKS gene was expected to result in the elimination of sulochrin biosynthesis. To perform the disruption by homologous recombination, a fragment of the emodin anthrone PKS gene first needed to be isolated. Analysis of several reported fungal PKS amino acid sequences has identified three subfamilies of related sequences (called the Patulin subfamily, the Pigment subfamily, and the Reduction subfamily). PCR primers specific for the Pigment subfamily (of which the emodin anthrone PKS is expected to belong) were used to isolate a fragment of a novel PKS gene from A. terreus. Targeted gene disruption identifies the novel gene fragment as that from the emodin anthrone PKS. Consequently, the gene disruption event eliminated the production of metabolites from the sulochrin biosynthetic pathway. PMID- 15129727 TI - Subcellular targeting of human interleukin-10 in plants. AB - The utility of plants for the production of a wide range of recombinant proteins is now clearly established. However, the challenge remains to produce these proteins at sufficient concentrations for extraction to be economically feasible. In this paper, we have investigated the ability of plant cells to accumulate the human interleukin-10 (IL-10) protein targeted to chloroplasts and mitochondria. We found that IL-10 accumulates in chloroplasts only if a 6 x His tag is added at the C-terminus of the protein. The hexapeptide may provide protection from degradation. Conversely, the IL-10 protein does not accumulate in mitochondria. Analysis of the chloroplast-targeted IL-10 protein revealed only monomeric IL-10 and limited biological activity in in vitro cell assays. PMID- 15129728 TI - High-level secretory production of recombinant bovine enterokinase light chain by Pichia pastoris. AB - Enterokinase (EC 3.4.21.9) is a serine proteinase with a specific digest sequence (Asp)4-Lys in the duodenum. Its high specificity for the recognition site makes enterokinase (EK) a useful tool for an in vitro cleavage of fusion proteins. In this work, an active bovine enterokinase light chain (EK(L)) was produced in secretory form by a recombinant strain of the methylotrophic yeast Pichia pastoris. The influences of methanol utilization phenotype of the host strain, induction pH, and carbon source on the recombinant production were studied. The production of recombinant EK(L) by Mut(s) strain was much higher than that by Mut+ strain. When inducted at pH 6.0, on a glycerol/methanol medium, the concentration of recombinant EK(L) (rEK(L)) reached 350 mg l(-1), which was 20 fold higher than that reported previously. The recombinant EK(L) was purified in a simple procedure on the anion exchange chromatography and 15 mg pure active EK(L) were obtained from 100 ml culture broth supernatant. The specific activity of purified rEK(L) was approximately 9000 u mg(-1). To facilitate purification and removal of rEKL after cleavage of fusion protein, the C-terminal His-tagged EK(L) (EK(L)/His) was also expressed in P. pastoris, and this His-tagged EK(L) exhibited a similar enzymatic activity to the untagged EK(L). PMID- 15129729 TI - Inactivation of alcohol dehydrogenase by piroxicam-derived radicals. AB - Alcohol dehydrogenase (ADH) was used as a marker molecule to clarify the mechanism of gastric mucosal damage as a side effect of using piroxicam. Piroxicam inactivated ADH during interaction of ADH with horseradish peroxidase and H2O2 (HRP-H2O2). The ADH was more easily inactivated under aerobic than anaerobic conditions, indicating participation by oxygen. Superoxide dismutase, but not hydroxyl radical scavengers, inhibited inactivation of ADH, indicating participation by superoxide. Sulfhydryl (SH) groups in ADH were lost during incubation of piroxicam with HRP-H2O2. Adding reduced glutathione (GSH) efficiently blocked ADH inactivation. Other SH enzymes, including creatine kinase and glyceraldehyde-3-phosphate dehydrogenase, were also inactivated by piroxicam with HRP-H2O2. Thus SH groups in the enzymes seem vulnerable to piroxicam activated by HRP-H2O2. Spectral change in piroxicam was caused by HRP-H2O2. ESR signals of glutathionyl radicals occurred during incubation of piroxicam with HRP H2O2 in the presence of GSH. Under anaerobic conditions, glutathionyl radical formation increased. Thus piroxicam free radicals interact with GSH to produce glutathionyl radicals. Piroxicam peroxyl radicals or superoxide, or both, seem to inactivate ADH. Superoxide may be produced through interaction of peroxyl radicals with H2O2. Thus superoxide dismutase may inhibit inactivation of ADH through reducing piroxicam peroxyl radicals or blocking interaction of SH groups with O2 , or both. Other oxicam derivatives, including isoxicam, tenoxicam and meloxicam, induced ADH inactivation in the presence of HRP-H2O2. PMID- 15129730 TI - Role of thioredoxin peroxidase in aging of stationary cultures of Saccharomyces cerevisiae. AB - A soluble protein from Saccharomyces cerevisiae acts as a peroxidase but requires a NADPH-dependent thioredoxin system and was named thioredoxin peroxidase (TPx). The role of TPx in aging of stationary cultures of S. cerevisiae was investigated in a wild-type strain and a mutant yeast strain in which the tsa gene that encodes TPx was disrupted by homologous recombination. The occurrence of oxidative stress during aging of stationary cultures of the yeast has been proposed. Comparison of 5-day-old (young) stationary cultures of S. cerevisiae and of cultures aged for 3 months (old) revealed decreased viability, increased generation of reactive oxygen species, modulation of cellular redox status, and increased cellular oxidative damage reflected by increased protein carbonyl content and lipid peroxidation. The magnitude of this stress was augmented in yeast mutant lacking TPx. These results suggest that TPx may play a direct role in cellular defense against aging of stationary cultures presumably, functioning as an antioxidant enzyme. PMID- 15129731 TI - Fish oil supplementation in pregnancy lowers F2-isoprostanes in neonates at high risk of atopy. AB - The anti-inflammatory properties of n-3 polyunsaturated fatty acids (n-3 PUFA) have suggested a potential role of these nutrients in dietary modification for prevention of allergic disease in early life. As oxidative stress is known to modify antigen presenting cell (APC) signalling and resulting immune responses, we examined the effects of maternal n-3 PUFA supplementation in pregnancy on markers of oxidative stress and APC function in neonates at high risk of allergy. Eighty-three pregnant atopic women were randomised to receive 4 g daily of either fish oil (n = 40) or olive oil (n = 43) capsules in a controlled trial from 20 weeks gestation until delivery. Plasma (cord blood) and urinary F2-isoprostanes were measured as markers of lipid peroxidation. Cord erythrocyte fatty acids and markers of APC function (HLA-DR expression and cytokine responses) were measured and related to levels of plasma F2-isoprostanes. Maternal fish oil supplementation lowered plasma (p < 0.0001) and urinary (p = 0.06) F2 isoprostanes. HLA-DR expression on APC was not different between the groups. In multiple regression analysis, 28.8% of the variance in plasma F2-isoprostanes was explained by positive relationships with erythrocyte arachidonic acid (AA) and monocyte HLA-DR expression and a negative relationship with erythrocyte eicosapentaenoic acid (EPA). This study shows that maternal supplementation with fish oil can attenuate neonatal lipid peroxidation. Clinical follow-up of these infants will help to determine if there are sustained effects on postnatal oxidative stress and expression of allergic disease. PMID- 15129732 TI - Inactivation of NADP+-dependent isocitrate dehydrogenase by lipid peroxidation products. AB - Membrane lipid peroxidation processes yield products that may react with proteins to cause oxidative modification. Recently, we demonstrated that the control of cytosolic and mitochondrial redox balance and oxidative damage is one of the primary functions of NADP+-dependent isocitrate dehydrogenase (ICDH) through to supply NADPH for antioxidant systems. When exposed to lipid peroxidation products, such as malondialdehyde (MDA), 4-hydroxynonenal (HNE) and lipid hydroperoxide, ICDH was susceptible to oxidative damage, which was indicated by the loss of activity and the formation of carbonyl groups. The structural alterations of modified enzymes were indicated by the change in thermal stability, intrinsic tryptophan fluorescence and binding of the hydrophobic probe 8-anilino 1-napthalene sulfonic acid. Upon exposure to 2,2'-azobis(2 amidinopropane) hydrochloride (AAPH), which induces lipid peroxidation in membrane, a significant decrease in both cytosolic and mitochondrial ICDH activities were observed in U937 cells. Using immunoprecipitation and immunoblotting, we were able to isolate and positively identify HNE adduct in mitochondrial ICDH from AAPH-treated U937 cells. The lipid peroxidation-mediated damage to ICDH may result in the perturbation of the cellular antioxidant defense mechanisms and subsequently lead to a prooxidant condition. PMID- 15129733 TI - The reactions of oxicam and sulfoanilide non steroidal anti-inflammatory drugs with hypochlorous acid: determination of the rate constants with an assay based on the competition with para-aminobenzoic acid chlorination and identification of some oxidation products. AB - Hypochlorous acid (HOCl) is an oxygen-derived species involved in physiological processes related to the defence of the organism that may cause adverse effects when its production is insufficiently controlled. In order to examine its reactivity with potential scavenging molecules from the non steroidal anti inflammatory drugs (NSAIDs) family, a competition assay based on para aminobenzoic acid (PABA) chlorination was developed. The original optimised in vitro fluorimetric procedure offered the possibility to determine rate constants (ks) for the reaction with HOCl in physiologically relevant conditions. The specificity of the system was improved by a liquid chromatography (LC) which allows the separation of the drugs and their oxidation products. After determination of the rate constant for PABA chlorination by HOCl (mean +/- SD in M(-1) s(-1): 4.3 +/- 0.3 x 10(3)), the applied mathematical model for a chemical competition permits to obtain linear curves from competition studies between several NSAIDs and PABA. Their slopes provided the following rate constants for the different studied drugs: tenoxicam: 4.0 +/- 0.7 x 10(3), piroxicam: 3.6 +/- 0.7 x 10(3), lornoxicam: 4.3 +/- 0.7 x 10(3), meloxicam: 1.7 +/- 0.3 x 10(4), nimesulide: 2.3 +/- 0.6 x 10(2). Meloxicam therefore reacted significantly faster than the other oxicams and nimesulide, which is the weakest scavenger of the studied series. The identification of some of the oxidation products by NMR or MS permitted to explore the reaction mechanism and to examine some aspects of the structure/activity relationships for the molecules of the same chemical family. PMID- 15129734 TI - Detection of superoxide and peroxynitrite in model systems and mitochondria by the luminol analogue L-012. AB - In the present study we investigated the specificity and sensitivity of the chemiluminescence (CL) dye and luminol analogue 8-amino-5-chloro-7 phenylpyrido[3,4-d]pyridazine-1,4-(2H,3H) dione (L-012) to detect reactive oxygen species (ROS) such as superoxide, peroxynitrite and hydrogen peroxide in cell free systems as well as in isolated mitochondria. The results obtained by L-012 were compared with other CL substances such as luminol, lucigenin, coelenterazine and the fluorescence dye dihydroethidine. The results indicate that the L-012 derived chemiluminescence induced by superoxide from hypoxanthine/xanthine oxidase (HX/XO) or by 3-morpholino sydnonimine (SIN-1)-derived peroxynitrite largely depends on the incubation time. Irrespective of the experimental conditions, L-012-derived CL in response to HX/XO and SIN-1 was 10-100 fold higher than with other CL dyes tested. In a cell-free system, authentic peroxynitrite yielded a higher L-012-enhanced CL signal than authentic superoxide and the superoxide-induced signal in cell-free as well as isolated mitochondria increased in the presence of equimolar concentrations of nitrogen monoxide (NO). The superoxide signal/background ratio detected by L-012-enhanced CL in isolated mitochondria with blocked respiration was 7 fold higher than that obtained by the superoxide sensitive fluorescence dye dihydroethidine. We conclude that L-012 derived CL may provide a sensitive and reliable tool to detect superoxide and peroxynitrite formation in mitochondrial suspensions. PMID- 15129735 TI - Angeli's salt and spinal motor neuron injury. AB - Nitroxyl anion or its conjugate acid (NO-/HNO) and nitric oxide (NO) may both have pro-oxidative and cytotoxic properties. Superoxide dismutase (SOD) enzyme has been shown to convert reversibly HNO to NO. Mutations found in the SOD enzyme in some familial amyotrophic lateral sclerosis (ALS) patients affect redox properties of the SOD enzyme in a manner, which may affect the equilibrium between NO and HNO. Therefore, we studied the effects of HNO releasing compound, Angeli's salt (AS), on both motor and sensory functions after intrathecal administration in the lumbar spinal cord of a male rat. These functions were measured by rotarod, spontaneous activity, paw- and tail-flick tests. In addition, we compared the effect of AS to NO releasing papanonoate, old AS solution and sulphononoate in the motor performance test. The effect of intrathecal delivery of AS on the markers of the spinal cord injury and oxidative/nitrosative stress were further studied. RESULTS: Freshly prepared AS (5 or 10 micromol), but not papanonoate, caused a marked decrease in the rotarod performance 3-7 days after the intrathecal administration. The peak motor deficiency was noted 3 days after AS (5 micromol) delivery. Old, degraded, AS solution and nitrous oxide releasing sulphononoate did not decrease motor performance in the rotarod test. AS did not affect the sensory stimulus evoked responses as measured by the paw-flick and tail-flick tests. Immunohistological examination revealed that AS caused injury related changes in the expression of glial fibrillary acidic protein (GFAP), fibroblast growth factor (FGF-2) and laminins in the spinal cord. Moreover, AS increased nitrotyrosine immunoreactivity in the spinal motor neurons. Therefore, we conclude that AS, but not NO releasing papanonoate, causes motor neuron injury but does not affect the function of sensory nerves in behavioural tests. PMID- 15129736 TI - Effects of pretreatment with a xanthine oxidase inhibitor on free radical levels during carotid endarterectomy. AB - OBJECTIVE: Free radicals contribute to the tissue damage caused by ischaemia reperfusion. The aim of the present study was to investigate whether preoperative antioxidant therapy (allopurinol) affects free radical levels in cerebral venous blood in connection with surgery for carotid artery stenosis. MATERIALS AND METHODS: Twenty-five patients were randomised into the study. Thirteen were controls and 12 were pretreated with allopurinol the day before surgery. Before, during and after surgery, blood samples were drawn from the ipsilateral jugular vein. Radical levels were measured using the spin trap technique ex vivo using OXANOH as the spin trap. Multivariate statistics were used with Principal Component Analysis and Partial Least Square regression analysis. RESULTS: Radical levels increased with diabetes, high leukocyte count, high creatinine and a high degree of contralateral stenosis. Radical levels decreased with high age, blood pressure, collateral circulation as well as operation for left-side carotid artery stenosis. After pretreatment with allopurinol, several of the relationships noted in the control group were eliminated, i.e. leukocyte count, side of operation, Betapred pretreatment and collateral circulation. CONCLUSIONS: Radical levels can be determined in connection with surgery for carotid artery stenosis using an ex vivo spin trap method. With preoperative antioxidant therapy the relationships between enhanced radical levels and clinical data, as seen in control subjects, disappeared. This might indicate a beneficial effect of preoperative pretreatment with antioxidants. PMID- 15129737 TI - Oxygen-copper (II) interplay in the repair of semi-oxidized urate by quercetin bound to human serum albumin. AB - The 1:1 complex of copper (II) and human serum albumin (HSA) slowly reacts with radiolytically generated *O2- radical-anion at a rate constant of 6.1 x 10(6) M( 1) s(-1). Absorbance and fluorescence spectroscopies demonstrate that addition of an equimolar portion of quercetin (QH2) to the solution of the copper (II)-HSA complex induces a relocalization of the copper resulting in a ternary copper (II) QH2-HSA complex. This form of quercetin slowly oxidizes in air-saturated solutions. A 10-fold excess urate, a plasma antioxidant, cannot displace copper (II) bound to HSA. In N2O-saturated solutions the ternary complex form of QH2 can repair the urate radical with a rate constant of 2.7 x 10(6) M(-1) s(-1) by an electron transfer reaction similar to that observed in the absence of copper (II). In O2-saturated solutions and in the absence of copper, HSA-bound QH2 fails to repair the urate radical because of the fast competitive reaction of *O2- with urate radicals. However, addition of equimolar copper (II) restores the electron transfer from QH2 to the urate radical. These contrasting results are tentatively explained either by an enhanced reactivity of copper (II) with *O2- in the ternary complex or by direct production of quercetin radicals via a copper catalyzed reduction of the *O2- radicals by QH2. PMID- 15129738 TI - Manganese complexes of curcumin analogues: evaluation of hydroxyl radical scavenging ability, superoxide dismutase activity and stability towards hydrolysis. AB - In order to improve the antioxidant property of curcumin and its analogue, diacetylcurcumin, manganese was incorporated into the structures in order to enhance superoxide dismutase (SOD) activity. Manganese (Mn) complexes of curcumin (CpCpx) and diacetylcurcumin (AcylCpCpx) were synthesized and firstly investigated for SOD activity and hydroxyl radical (HO*) scavenging ability. SOD activity was evaluated by both the nitroblue tetrazolium (NBT) reduction assay and electron paramagnetic resonance (EPR) with 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a spin trapping agent. CpCpx and AcylCpCpx inhibited the NBT reduction and decreased the DMPO/OOH adduct much greater than corresponding antioxidants or ligands, with IC50 values of 29.9 and 24.7 microM (NBT), and 1.09 and 2.40 mM (EPR), respectively. For EPR, potassium superoxide (KO2) was used as a source of O2- where qualitative results suggested that CpCpx and AcylCpCpx were SOD mimics, which catalyze the conversion of O2- to dioxygen and hydrogen peroxide (H2O2). Additionally, CpCpx and AcylCpCpx exhibited the great inhibition of DMPO/OH adduct formation with an IC50 of 0.57 and 0.37mM, respectively, which were comparable to that of curcumin (IC50 of 0.64 mM), indicating that both Mn complexes are also an effective HO* scavenger. The stability against hydrolysis in water, various buffers and human blood/serum was carried out in vitro. It was found that both Mn complexes were pH and salt concentration dependent, being more stable in basic pH. In the human blood/serum test, CpCpx was more stable against hydrolysis than AcylCpCpx with about 10 and 20% of free Mn2+ releasing, respectively. PMID- 15129739 TI - Evaluation of the antioxidant properties of propofol and its nitrosoderivative. comparison with homologue substituted phenols. AB - Propofol (2,6-diisopropylphenol), some substituted phenols (2,6-dimethylphenol and 2,6-ditertbutylphenol) and their 4-nitrosoderivatives have been compared for their scavenging ability towards 1,1-diphenyl-2-picrylhydrazyl and for their inhibitory action on lipid peroxidation. These products were also compared to the classical antioxidants butylated hydroxytoluene and butylated hydroxyanisole. When measuring the reactivity of the various phenolic derivatives with 1,1 diphenyl-2-picrylhydrazyl the following order of effectiveness was observed: butylated hydroxyanisole > propofol > 2,6-dimethylphenol > 2,6-di-tertbutylphenol > butylated hydroxytoluene. In cumene hydroperoxide-dependent microsomal lipid peroxidation, propofol acts as the most effective antioxidant, while butylated hydroxyanisole, 2,6-di-tertbutylphenol and butylated hydroxytoluene exhibit a rather similar effect, although lower than propofol. In the iron/ascorbate dependent lipid peroxidation propofol, at concentrations higher than 10 microM, exhibits antioxidant properties comparable to those of butylated hydroxytoluene and butylated hydroxyanisole, 2,6-Dimethylphenol is scarcely effective in both lipoperoxidative systems. The antioxidant properties of the various molecules depend on their hydrophobic characteristics and on the steric and electronic effects of their substituents. However, the introduction of the nitroso group in the 4-position almost completely removes the antioxidant properties of the examined compounds. The nitrosation of the aromatic ring of antioxidant molecules and the consequent loss of antioxidant capacity can be considered a condition potentially occurring in vivo since nitric oxide and its derivatives are continuously formed in biological systems. PMID- 15129740 TI - Endogenous and dietary indoles: a class of antioxidants and radical scavengers in the ABTS assay. AB - Indoles are very common in the body and diet and participate in many biochemical processes. A total of twenty-nine indoles and analogs were examined for their properties as antioxidants and radical scavengers against 2,2'-Azino-bis(3 ethylbenzthiazoline-6-sulfonic acid) ABTS*+ radical cation. With only a few exceptions, indoles reacted nonspecifically and quenched this radical at physiological pH affording ABTS. Indoleamines like tryptamine, serotonin and methoxytryptamine, neurohormones (melatonin), phytohormones (indoleacetic acid and indolepropionic acid), indoleamino acids like L-tryptophan and derivatives (N acetyltryptophan, L-abrine, tryptophan ethyl ester), indolealcohols (tryptophol and indole-3-carbinol), short peptides containing tryptophan, and tetrahydro-beta carboline (pyridoindole) alkaloids like the pineal gland compound pinoline, acted as radical scavengers and antioxidants in an ABTS assay-measuring total antioxidant activity. Their trolox equivalent antioxidant capacity (TEAC) values ranged from 0.66 to 3.9 mM, usually higher than that for Trolox and ascorbic acid (1 mM). The highest antioxidant values were determined for melatonin, 5 hydroxytryptophan, trp-trp and 5-methoxytryptamine. Active indole compounds were consumed during the reaction with ABTS*+ and some tetrahydropyrido indoles (e.g. harmaline and 1-methyl-1,2,3,4-tetrahydro-beta-carboline-3-carboxylic acid ethyl ester) afforded the corresponding fully aromatic beta-carbolines (pyridoindoles), that did not scavenge ABTS*+. Radical scavenger activity of indoles against ABTS*+ was higher at physiological pH than at low pH. These results point out to structural compounds with an indole moiety as a class of radical scavengers and antioxidants. This activity could be of biological significance given the physiological concentrations and body distribution of some indoles. PMID- 15129741 TI - Reading music modifies spatial mapping in pianists. AB - We used a novel musical Stroop task to demonstrate that musical notation is automatically processed in trained pianists. Numbers were superimposed onto musical notes, and participants played five-note sequences by mapping from numbers to fingers instead of from notes to fingers. Pianists' reaction times were significantly affected by the congruence of the note/number pairing. Nonmusicians were unaffected. In a nonmusical analogue of the task, pianists and nonmusicians showed a qualitative difference on performance of a vertical-to horizontal stimulus-response mapping task. Pianists were faster when stimuli specifying a leftward response were presented in vertically lower locations and stimuli specifying a rightward response were presented in vertically higher locations. Nonmusicians showed the reverse pattern. No group differences were found on a task that required horizontal-to-horizontal mappings. We suggest that, as a result of learning to read and play keyboard music, pianists acquire vertical-to-horizontal visuomotor mappings that generalize outside the musical context. PMID- 15129742 TI - A response time model for judging order relationship between two symbolic stimuli. AB - The response time to judge the order relationship between two symbolic stimuli is frequently modeled as the time spent in a (constant-rate) accumulative sampling process until a threshold is reached. We will show that empirical descriptions of observed effects in number comparisons suggest an accrual process that reaches the threshold at an exponential rate. The model accrual equations and stopping conditions have an immediate interpretation in terms of a simple quantitative connectionist network. The encoded stimuli and thresholds are inputs to the network. The former are considered to result from the participant's learning history, and the latter modulate the rearrangement of the network parts; each arrangement models a different task. We have found a good correlation between model predictions and other authors' experimental data, both in number comparisons and in experiments in which the ordering of the symbolic stimuli has been artificially induced. Incorrect answers are discussed, and predictions are compared with data. We will explore differences and similarities with other approaches, such as random walk and the symbolic comparison model. In a limit case, our model becomes identical to the discriminability model. PMID- 15129743 TI - Probe distractors can influence negative priming by perceptual grouping. AB - Negative priming occurs when a response is slower to a target that was a previously ignored stimulus. According to a computational model of inhibition, negative priming results from relative activation between a target and a distractor in probe trials. Thus, the degree of competition in the probe trial should influence negative priming. The results from three experiments supported this hypothesis. Probe distractors grouped with a target by a rectangle produced a larger negative priming effect than did distractors isolated from the target. Moreover, neither prime-probe similarity nor perceptual interference could explain the grouping effect. The results suggest that perceptual mechanisms can influence negative priming via a race in the probe trial. PMID- 15129744 TI - Supporting the attentional momentum view of IOR: is attention biased to go right? AB - Inhibition of return (IOR) refers to the finding that individuals are slower to respond to a target presented at a previously attended location than they are to respond to a target presented at a novel location (Posner & Cohen, 1984). The attentional momentum theory is a recent view of how attention moves around the environment, and it provides an account for the IOR effect that does not rely on an inhibitory mechanism (Pratt, Spalek, & Bradshaw, 1999). The present paper supports the attentional momentum viewpoint in two ways: first, by replicating the finding that reaction times to targets at the uncued locations are not all the same (Pratt et al., 1999) and second, by showing that responses made to all locations on the cued side of fixation, and not just to the locations that attention had previously traversed, are slower than are responses made to locations on the opposite side of fixation. We also demonstrate that there is a directional bias to the IOR effect that results in the effect's being larger when attention moves in a left-to-right manner. PMID- 15129745 TI - Recognizing a plane-misoriented view of a familiar object is not influenced by the ease of specifying the main axis of elongation of that object. AB - Participants saw three versions of pictures of familiar objects: the original unaltered (axis-normal) pictures, axis-extended pictures in which the main axes of the axis-normal pictures were elongated, and axis-switched pictures in which objects that were originally horizontally elongated were depicted as vertically elongated and vice versa. Relative to axis-normal pictures, axis extension aided decisions about whether the picture of the object was wide or tall, and axis switching hindered these decisions for both upright and plane-misoriented views. Nevertheless, although these axis manipulations clearly influenced decisions about the location of the object's main axis of elongation, axis-switched pictures were no harder to name than axis-extended pictures. Changing the depicted main axis of elongation by axis switching and axis extension did not influence object recognition in itself, whether for upright or for plane misoriented views. This suggests that specifying the main axis of elongation of an object does not play an important role in the orientation-sensitive processes involved in identifying plane-misoriented views of that object. PMID- 15129746 TI - Voluntary stuttering suppresses true stuttering: a window on the speech perception-production link. AB - In accord with a proposed innate link between speech perception and production (e.g., motor theory), this study provides compelling evidence for the inhibition of stuttering events in people who stutter prior to the initiation of the intended speech act, via both the perception and the production of speech gestures. Stuttering frequency during reading was reduced in 10 adults who stutter by approximately 40% in three of four experimental conditions: (1) following passive audiovisual presentation (i.e., viewing and hearing) of another person producing pseudostuttering (stutter-like syllabic repetitions) and following active shadowing of both (2) pseudostuttered and (3) fluent speech. Stuttering was not inhibited during reading following passive audiovisual presentation of fluent speech. Syllabic repetitions can inhibit stuttering both when produced and when perceived, and we suggest that these elementary stuttering forms may serve as compensatory speech gestures for releasing involuntary stuttering blocks by engaging mirror neuronal systems that are predisposed for fluent gestural imitation. PMID- 15129747 TI - Right-left prevalence effect with horizontal and vertical effectors. AB - We investigated the right-left prevalence effect in spatial compatibility tasks by assessing subjects' performance on a two-dimensional task in which both the horizontal and vertical spatial dimensions were task relevant. Two experiments were performed, in which stimulus-response mappings were one-dimensional (Experiment 1) and two-dimensional (Experiment 2). The subjects responded by using either horizontal or vertical effectors to stimuli appearing in four possible locations. With the one-dimensional mapping, the spatial compatibility effect was present only in the dimension relevant to the mapping. With the two dimensional mapping, the horizontal compatibility effect was always present, whereas the vertical compatibility effect was present only when vertical effectors were used. This pattern of results indicates that horizontal coding takes place with either horizontal or vertical effectors, whereas vertical coding takes place only when vertical effectors are used. PMID- 15129748 TI - Endogenous temporal orienting of attention in detection and discrimination tasks. AB - Endogenous temporal-orienting effects were studied using a cuing paradigm in which the cue indicated the time interval during which the target was most likely to appear. Temporal-orienting effects were defined by lower reaction times (RTs) when there was a match between the temporal expectancy for a target (early or late) and the time interval during which the target actually appeared than when they mismatched. Temporal-orienting effects were found for both early and late expectancies with a detection task in Experiment 1. However, catch trials were decisive in whether temporal-orienting effects were observed in the early expectancy condition. No temporal-orienting effects were found in the discrimination task. In Experiments 2A and 2B, temporal-orienting effects were observed in the discrimination task; however, they were larger when temporal expectancy was manipulated between blocks, rather than within blocks. PMID- 15129749 TI - Parafoveal processing of prefixed words during eye fixations in reading: evidence against morphological influences on parafoveal preprocessing. AB - Two eye movement experiments were conducted to investigate whether the morphological constituents of a prefixed word influenced early word processing when English was read. Participants read sentences containing free-stem, bound stem, or pseudoprefixed words, and the availability of the prefixed word morphemes in the parafovea was manipulated. Although preview benefit was greatest for the entire word, there was evidence that subsequent word processing was facilitated in both the prefix-only and the stem-only conditions. This effect was not influenced by word type. There was no evidence that morphological preprocessing occurred when morphological information was available in the parafovea. In addition, reading times for the target word did not differ for the different word types. Thus, there was no evidence to suggest that morphological constituents influence early word processing during reading. It is possible that morphological effects tend to be obscured when examined within a sentence context (although this phenomenon may be specific to English). PMID- 15129750 TI - Selective learning of spatial configuration and object identity in visual search. AB - To conduct an efficient visual search, visual attention must be guided to a target appropriately. Previous studies have suggested that attention can be quickly guided to a target when the spatial configurations of search objects or the object identities have been repeated. This phenomenon is termed contextual cuing. In this study, we investigated the effect of learning spatial configurations, object identities, and a combination of both configurations and identities on visual search. The results indicated that participants could learn the contexts of spatial configurations, but not of object identities, even when both configurations and identities were completely correlated (Experiment 1). On the other hand, when only object identities were repeated, an effect of identity learning could be observed (Experiment 2). Furthermore, an additive effect of configuration learning and identity learning was observed when, in some trials, each context was the relevant cue for predicting the target (Experiment 3). Participants could learn only the context that was associated with target location (Experiment 4). These findings indicate that when multiple contexts are redundant, contextual learning occurs selectively, depending on the predictability of the target location. PMID- 15129751 TI - Frame effects in visual search for line orientation. AB - In the present study, we investigated the effects of surrounding frames on visual search for line orientation. Every line item presented in the display was surrounded by a square frame of identical size and orientation. The orientations of the frames, as well as those of the target and distractor lines, were either vertical or tilted. In six experiments, the surrounding frames caused substantial changes in search efficiency for vertical targets and for tilted targets. The search asymmetry between the two types of targets was reversed when the frame was tilted at the same angle as the tilted line. Several variations of the frames (a pair of parallel lines, squares with gaps, and squares with circular contours inside) also changed search efficiency significantly. Taken together, these results imply that three different sources contribute to frame effects: distractor roles played by the frame components, orientation contrast from the frame contour, and interference in local orientation processing (Hayward & Burke, 2000). Implications of the present findings are discussed in reference to rod-and frame effects (Witkin & Asch, 1948) and to the effects of a large frame surrounding an entire display of lines (Marendaz, 1998; Treisman, 1985). PMID- 15129752 TI - Time perception with and without a concurrent nontemporal task. AB - Prospective time estimates were obtained from human subjects for stimulus durations ranging from 2 to 23 sec. Presence and absence of a concurrent nontemporal task was manipulated within subjects in three experiments. In addition, location of the task within temporal reproduction trials and psychophysical method were varied between groups in Experiments 2 and 3, respectively. For long-duration stimuli, the results of all three experiments conformed to results in the literature, showing a decrease in perceived duration under concurrent task conditions, in accord with attentional resource allocation models of timing. The effects of task location and psychophysical method on time estimates were also compatible with this analysis. However, psychophysical functions obtained under task conditions were fit well by power functions, an outcome that would not be anticipated on the basis of attention theory. The slopes of the functions under no-task conditions were steeper than those under task conditions. The data support the perceptual hypothesis that different sources of sensory input mediate timing under task and no-task conditions. PMID- 15129753 TI - The visual and haptic perception of natural object shape. AB - In this study, we evaluated observers' ability to compare naturally shaped three dimensional (3-D) objects, using their senses of vision and touch. In one experiment, the observers haptically manipulated 1 object and then indicated which of 12 visible objects possessed the same shape. In the second experiment, pairs of objects were presented, and the observers indicated whether their 3-D shape was the same or different. The 2 objects were presented either unimodally (vision-vision or haptic-haptic) or cross-modally (vision-haptic or haptic vision). In both experiments, the observers were able to compare 3-D shape across modalities with reasonably high levels of accuracy. In Experiment 1, for example, the observers' matching performance rose to 72% correct (chance performance was 8.3%) after five experimental sessions. In Experiment 2, small (but significant) differences in performance were obtained between the unimodal vision-vision condition and the two cross-modal conditions. Taken together, the results suggest that vision and touch have functionally overlapping, but not necessarily equivalent, representations of 3-D shape. PMID- 15129754 TI - Audiovisual speech facilitates voice learning. AB - In this research, we investigated the effects of voice and face information on the perceptual learning of talkers and on long-term memory for spoken words. In the first phase, listeners were trained over several days to identify voices from words presented auditorily or audiovisually. The training data showed that visual information about speakers enhanced voice learning, revealing cross-modal connections in talker processing akin to those observed in speech processing. In the second phase, the listeners completed an auditory or audiovisual word recognition memory test in which equal numbers of words were spoken by familiar and unfamiliar talkers. The data showed that words presented by familiar talkers were more likely to be retrieved from episodic memory, regardless of modality. Together, these findings provide new information about the representational code underlying familiar talker recognition and the role of stimulus familiarity in episodic word recognition. PMID- 15129755 TI - Clonal population of adult stem cells: life span and differentiation potential. AB - Adult stem cells derived from bone marrow, connective tissue, and solid organs can exhibit a range of differentiation potentials. Some controversy exists regarding the classification of mesenchymal stem cells as bona fide stem cells, which is in part derived from the limited ability to propagate true clonal populations of precursor cells. We isolated putative mesenchymal stem cells from the connective tissue of an adult rat (rMSC), and generated clonal populations via three rounds of dilutional cloning. The replicative potential of the clonal rMSC line far exceeded Hayflick's limit of 50-70 population doublings. The high capacity for self-renewal in vitro correlated with telomerase activity, as demonstrated by telomerase repeat amplification protocol (TRAP) assay. Exposure to nonspecific differentiation culture medium revealed multilineage differentiation potential of rMSC clones. Immunostaining confirmed the appearance of mesodermal phenotypes, including adipocytes possessing lipid-rich vacuoles, chondrocytes depositing pericellular type II collagen, and skeletal myoblasts expressing MyoD1. Importantly, the spectrum of differentiation capability was sustained through repeated passaging. Furthermore, serum-free conditions that led to high-efficiency smooth muscle differentiation were identified. rMSCs plated on collagen IV-coated surfaces and exposed to transforming growth factor-beta1 (TGF beta1) differentiated into a homogeneous population expressing alpha-actin and calponin. Hence, clonogenic analysis confirmed the presence of a putative MSC population derived from the connective tissue of rat skeletal muscle. The ability to differentiate into a smooth muscle cell (SMC) phenotype, combined with a high proliferative capacity, make such a connective tissue-derived MSC population ideal for applications in vascular tissue construction. PMID- 15129756 TI - Transplantation of adipose tissue-derived stromal cells increases mass and functional capacity of damaged skeletal muscle. AB - The regenerating skeletal muscle environment is capable of inducing uncommitted progenitors to terminally differentiate. The aim of this work was to determine whether adipose tissue-derived stromal cells were able to participate in muscle regeneration and to characterize the effect on muscle mass and functional capacities after transplantation of these cells. Adipose tissue stromal cells labeled with Adv cyto LacZ from 3-day-old primary cultures (SVF1) were autotransplanted into damaged tibialis anterior muscles. Fifteen days later, beta galactosidase staining of regenerated fibers was detected, showing participation of these cells in muscle regeneration. Two months after SVF1 cell transfer, muscles were heavier, showed a significantly larger fiber section area, and developed a significantly higher maximal force compared with damaged control muscles. These results are similar to those previously obtained after satellite cell transplantation. However, SVF1 transfer also generated a small amount of adipose tissue localized along the needle course. To minimize these adipose contaminants, we transferred cells from 7-day-old secondary cultures of the SVF1, containing only a small proportion of already engaged preadipocytes (SVF2). Under these conditions, no adipose tissue was observed in regenerated muscle but there was also no effect on muscle performances compared with damaged control muscles. This result provides further evidence for the existence of progenitor cells in the stromal fraction of freshly isolated adipose tissue cells, which, under our conditions, keep some of their pluripotent properties in primary cultures. PMID- 15129757 TI - Intraspinal transplantation of CD34+ human umbilical cord blood cells after spinal cord hemisection injury improves functional recovery in adult rats. AB - The present study was designed to compare the functional outcome of the intraspinal transplantation of CD34+ human umbilical cord blood (CB) cells with that of human bone marrow stromal (BMS) cells in adult rats with spinal cord injury. Sixty adult Wistar rats were subjected to left spinal cord hemisection, and then divided into three groups randomly. The control group received an injection of PBS without cells, while the two other groups of rats received a transplantation of 5 x 10(5) CD34+ CB or BMS cells, respectively. Functional outcome was measured using the modified Tarlov score at days 1, 7, 14, 21, and 28 after transplantation. A statistically significant improvement in functional outcome and survival rate in the experimental groups of rats was observed compared with the control group. Rats that received CD34+ CB cells achieved a better improvement in functional score than those that received BMS cells at days 7 and 14 after transplantation. Histological evaluation revealed that bromodeoxyuridine (BrdU)-labeled CD34+ CB and BMS cells survived and migrated into the injured area. Some of these cells expressed glial fibriliary acidic protein (GFAP) or neuronal nuclear antigen (NeuN). Our data demonstrate for the first time that intraspinal transplantation of human CD34+ CB cells provides benefit in function recovery after spinal cord hemisection in rats and suggest that CD34+ CB cells may be an excellent choice of cells as routine starting material of allogenic and autologous transplantations for the treatment of spinal cord injury. PMID- 15129758 TI - Transplanted hNT cells ("LBS neurons") in a rat model of huntington's disease: good survival, incomplete differentiation, and limited functional recovery. AB - A variety of immortalized cell lines have been proposed to exhibit sufficient phenotypic plasticity to allow them to replace primary embryonic neurons for restorative cell transplantation. In the present experiments we evaluate the functional viability of one particular cell line, the hNT cells developed by Layton Bioscience, to replace lost neurons and alleviate asymmetrical motor deficits in a unilateral excitotoxic lesion model of Huntington's disease. Because the grafts involved implantation of human-derived cells into a rat host environment, all animals were immunosuppressed. Cyclosporin A and FK-506 were similar in providing effective immunoprotection of the hNT xenografts, and whereas the lesions induced a marked inflammatory response in the host brain, this was not exacerbated by the presence of xenograft cells. The presence of grafted cells was determined with the human-specific antigen HuNu, and good graft survival was demonstrated in almost all animals up to the longest survival examined, 16 weeks posttransplantation. Although the cells exhibited progressively greater maturation and differentiation at 10-day, 4- and 16-week time points, staining for the mature neuronal marker NeuN was at best very weak, and we were unable to detect unequivocal staining with any markers of mature striatal phenotype, including DARPP-32, calbindin, parvalbumin, choline acetyl transferase, or NADPH diaphorase (with in all cases positive control provided by good staining on the intact contralateral side of the brain). Nor were we able to detect any differences between rats with lesions alone and rats with grafts in the contralateral motor deficits exhibited in a test of skilled paw reaching or cylinder placing. These results suggest that further and more extensive studies should be undertaken to assess whether hNT neurons can show more extensive and appropriate maturation and be associated with recovery in appropriate behavioral models, before they may be considered a suitable replacement for primary embryonic cells for clinical application in Huntington's disease. PMID- 15129759 TI - Natural antibodies prevent in vivo transmission of porcine islet-derived endogenous retrovirus to human cells. AB - The discovery of porcine endogenous retroviruses (PERV) has raised concerns regarding the safety of porcine xenotransplantation. However, transmission of PERV had not been observed in humans exposed to porcine tissue. We examined whether PERV derived from porcine pancreatic islet cells could infect human cells in vivo and the role of natural antibodies in inhibiting PERV infection. In vivo infective potential of PERV was studied in SCID mice reconstituted with human peripheral blood leucocytes. Porcine islets were transplanted under the kidney capsule. PERV infection was determined by analyzing PERV gene expression in graft infiltrating lymphocytes (GIL) harvested 21 days posttransplantation. Mice were administered normal human serum prior to and 2 days posttransplantation to study their role in protection of human cells against PERV infection. PERV genes were expressed in all porcine tissues examined, including purified porcine islets. PERV expression was detected in GILs from three of five human-SCID mice. Administration of human serum blocked PERV infection in GILs in five of five human-SCID mice. These results indicate that PERV from porcine islets can infect human cells in vivo. Normal human serum blocks transmission of retrovirus in vivo, suggesting that natural xenoreactive antibodies can prevent PERV infection. PMID- 15129760 TI - Polyphenol, an extract of green tea, increases culture recovery rates of isolated islets from nonhuman primate pancreata and marginal grade human pancreata. AB - Investigations indicate that an extract of green tea, polyphenol, can significantly increase the culture survival rate of rat islets without deteriorating their functionality. In this study, we examined the effect of adding polyphenol to islets isolated from human pancreata and nonhuman primate pancreata. Islets were isolated from human pancreata that did not meet criteria for clinical transplantation (n = 6) and from nonhuman primate pancreata (n = 5). The islets were cultured in CMRL-1066 + 10% FCS with the addition of 0, 30, 60, 125, 250, or 500 microg/ml of polyphenol. After 24 or 48 h of culture, islet yield, viability, purity, morphology, and stimulation index was assessed. RT-PCR and Western blot analysis were also performed to assess the expression levels of the apoptotic related genes, Bcl-2 and BAX. After 24 h of culture, islet yields were significantly higher in cultures supplemented with 30-250 microg/ml of polyphenol than in cultures without polyphenol. After 48 h of culture, significant differences in islet numbers were observed with polyphenol concentrations of 125 microg/ml (p < 0.01) and 250 microg/ml (p < 0.01). However, no significant differences were noted in islet viability, purity, morphology, and stimulation index at each time point with or without polyphenol. RT-PCR and Western blot analysis of the islets indicated that Bcl-2 levels increased by 2.5 fold and BAX levels decreased by twofold in cultures supplemented with polyphenol. This resulted in BAX/Bcl-2 ratios that were lower in polyphenol supplemented cultures than with control cultures. Polyphenol increases culture recovery rates by precluding islet apoptosis. PMID- 15129761 TI - Endogenous pancreatic enzyme activity levels show no significant effect on human islet isolation yield. AB - Despite advances in human islet isolation, islet yield remains inconsistent and unreliable. In recent studies, it has been suggested that serine proteases, in particular trypsin, have been shown to have a damaging effect on islet yield. This study evaluated enzyme activity levels throughout 42 human islet isolation procedures. Trypsin, chymotrypsin, and elastase activity was determined spectrophotometrically using suitable chromophoric substrates. The results of the islet isolations were rated as successful (n = 19) or unsuccessful (n = 23) based on the islet yield and functionality. The enzyme activity profiles of the isolations were compared. No significant differences in donor-related variables were found in this study. However, in the successful isolations, a significantly greater amount (85.6 +/- 1.9%; p = 0.0017) of the pancreas was digested in a significantly shorter digestion time (19.7 +/- 0.6 min; p = 0.0054) compared with 74.8 +/- 2.5% of digested tissue in 22.6 +/- 0.7 min in the poor isolations. This study showed no significant effect of serine protease levels on the outcome of islet isolations, regardless of enzyme inhibitor supplementation. These data suggest that serine protease activity does not sufficiently affect islet yield. However, the data show that the most successful human islet isolations are achieved when the maximum amount of tissue is digested in the shortest amount of time. This suggests that further understanding of the isolation process should focus on the role of the collagenase digestion solution in the dissociation of the endocrine-exocrine tissue connection. PMID- 15129762 TI - Fate of a chimeric joint construct in an ectopic site in SCID mice. AB - This study examines the use of a devitalized biological knee as a scaffold for repopulation with chondrocytes and tests the hypothesis that the devitalized scaffold would become repopulated with the foreign chondrocytes when placed in a suitable environment. Chimeric knee constructs were engineered in vitro and their ectopic in vivo fate was examined in SCID mice. The constructs were made by applying porous collagen sponges that contained viable bovine articular chondrocytes to shaved articular surfaces of devitalized embryonic chick knees. The chimeric joints were cultured for 1 week and were subsequently transplanted into dorsal subcutaneous pouches of 5-week-old mice. Specimens were prepared for histological analysis at 1, 3, 6, or 8 weeks after transplantation. Controls included empty collagen sponges, collagen sponges seeded with viable bovine chondrocytes, and devitalized chick knees without collagen sponge inserts. One week after in vitro incubation of the constructs, the porous collagen sponges with viable bovine chondrocytes were adherent to the shaved articular surfaces of the devitalized chick joints. There was abundant metachromatic neomatrix around the chondrocytes in the collagen sponges. During maintenance of the constructs in vivo, the chimeric joints exhibited dramatic changes. Bovine chondrocytes proliferated in the collagen sponges and formed abundant new matrix. Bovine chondrocytes migrated into preexisting chick cartilage canals at 1 week. Subsequently, bovine chondrocytes invaded the matrix of the devitalized chick knees. Bovine neocartilage obliterated the interface between the collagen sponge and the devitalized chick cartilage. With time in vivo, the bovine neocartilage expanded and replaced the chick matrix. The devitalized cartilage appears to provide a framework for supporting chondrogenesis in a chimeric joint. PMID- 15129763 TI - Tissue-engineered grafts matured in the right ventricular outflow tract. AB - Autologous smooth muscle cell (SMC)-seeded biodegradable scaffolds could be a suitable material to repair some pediatric right ventricular outflow tract (RVOT) cardiac anomalies. Adult syngenic Lewis rat SMCs (2 x 10(6)) were seeded onto a new biodegradable copolymer sponge made of epsilon-caprolactone-co-L-lactide reinforced with poly-L-lactide fabric (PCLA). Two weeks after seeding, the patch was used to repair a surgically created RVOT defect in an adult rat. At 8 weeks after implantation the spongy copolymer component was biodegraded, and SM tissue and extracellular matrices containing elastin fibers were present in the scaffolds. By 22 weeks more fibroblasts and collagen were present (p < 0.05). The number of capillaries in the grafts also increased (p < 0.001) between 8 and 22 weeks. The fibrous poly-L-lactide component of the PCLA scaffold remained. The 22 week grafts maintained their thickness and surface area in the RVOT. The SMCs prior to implantation were in a synthetic phenotype and developed in vivo into a more contractile phenotype. By 8 weeks the patches were endothelialized on their endocardial surfaces. Future work to increase the SM tissue and elastin content in the patch will be necessary before implantation into a pediatric large-animal model is tested. PMID- 15129764 TI - Assessment of different transfection parameters in efficiency optimization. AB - Achieving optimal transfection efficiency is the most critical step in overcoming the primary obstacle to success in nonviral-mediated gene therapy. Several transfection parameters were being examined including the effects of different types of transfection media, glucose concentration, reporter DNA concentration, and incubation time in lipotransfectant. Efficiency of transfection obtained was highest for Opti-MEM I (29 +/- 2.28%; p = 0.001) followed by M199 (24 +/- 1.54%; p = 0.009), both of which performed significantly better than DMEM (14 +/- 0.28%) as a transfection medium. The rate of transfection was affected by glucose levels in only DMEM with higher efficiency achieved using low glucose containing DMEM (17 +/- 0.38%) than its counterpart. Furthermore, transfection rate and cell viability were severely hampered by lengthened exposure to transfection complexes, leading to an overall mean efficiency of 5 +/- 0.87%. However, doubling the DNA content in the transfection mixture did not significantly change the mean rate of transfection in M199 medium (24 +/- 1.54% to 27 +/- 1.54%; p = 0.273). The overall range of mean efficiency acquired with our protocol under different transfection conditions was between 14% and 29%. Hopefully results from this study will further potential success in nonviral-mediated gene transfer. PMID- 15129765 TI - Long-term function of cryopreserved rat hepatocytes in a coculture system. AB - The goal of this study was to investigate postpreservation long-term function of cryopreserved primary rat hepatocytes using the hepatocyte/3T3-J2 fibroblast coculture system. The long-term function of thawed hepatocytes cocultured with fibroblasts was evaluated and compared with hepatocytes cultured without fibroblasts. Fresh isolated primary rat hepatocytes were frozen at a controlled rate (-1 degrees C/min) up to -80 degrees C, and then stored in liquid nitrogen for up to 90 days. Thawed hepatocytes were thereafter cocultured with 3T3-J2 murine fibroblasts and cocultivation was monitored for 14 days. The viability of fresh isolated hepatocytes was 91.4%, and that of cryopreserved hepatocytes was 82.1%. Cellular morphology and polarity, which were determined by the localization of actin filaments and connexin-32, were successfully maintained in cryopreserved hepatocytes following cryopreservation. Albumin and urea synthesis reached the maximum level and became stable after day 7 in coculture in both fresh and cryopreserved hepatocytes. Urea synthesis of cryopreserved hepatocytes was maintained 89.0% of nonfrozen fresh control, and albumin production of cryopreserved hepatocytes was 63.7% of control in coculture. Cytochrome P450 activity, which was measured by deethylation of ethoxyresorufin, was also maintained in cryopreserved hepatocytes at 88.6% of nonfrozen fresh control in coculture. The retention of synthetic and detoxification activities was verified to be well preserved during extended low-temperature storage (90 days). Both fresh control and cryopreserved hepatocytes cultured without fibroblast did not retain their synthetic and detoxification functions in long-term culture. These data illustrate that, through the utilization of our cryopreservation procedure, primary hepatocyte function was successfully maintained when placed into coculture configuration following thawing. PMID- 15129766 TI - Effects of methoxychlor, dieldrin and lindane on sea urchin fertilization and early development. AB - We have studied the effects of methoxychlor (MXC), dieldrin, and lindane on fertilization and early development of sea urchin egg. These organochlorine pesticides have often been found in polluted ground and water near agricultural sites, and have therefore been detected from time to time in the food chain and in drinking water. They have been reported to alter various reproduction functions in various animals including marine populations. We observed that the rate of fertilization decreased when the sperm was incubated with dieldrin or lindane. Treatment of eggs with each pesticide did not prevent fertilization, but increased the rate in polyspermy, delayed or blocked the first mitotic divisions, and altered early embryonic development. Moreover, all pesticides could alter several intracellular biochemical pathways that control first mitotic divisions and early development, including intracellular calcium homeostasis, MPF (mitosis promoting factor) activity and formation of the bipolar mitotic spindle. We found that lindane was the most potent of the three pesticides to alter all biochemical events. All these effects were observed at relatively high concentrations. However, bio-accumulation in sediments and aquatic organisms have been reported. Sea urchin eggs may then be in contact with very high concentrations of these pesticides in areas where these pesticides are not handled or stocked properly, and then develop into abnormal embryos. PMID- 15129767 TI - Sensitivity to cadmium along a salinity gradient in populations of the periwinkle, Littorina littorea, using time-to-death analysis. AB - In this study, we assessed the combined effect of Cd concentration and salinity, on Cd uptake and mortality rate of Littorina littorea, collected along a salinity and pollution gradient in the Western Scheldt estuary (The Netherlands). Animals kept at their field salinity levels were exposed to three Cd concentrations (i.e. 10, 40 and 320 microM), while animals kept in 10 microM of Cd were subjected to five salinity treatments (i.e. 15, 20, 25, 30 and 35 per thousand). Mortality was recorded every 24h and Cd body burdens were measured with ICP-AES. Time-to-death data were analysed via Cox proportional hazard models, including the co-variates "site-Cd treatment" in the Cd experiment and "site-salinity treatment" in the salinity experiment. "Cd-treatment" and "field-salinity" affected mortality rates significantly in the Cd experiment, such that the mortality risk increased by 2.3 times when salinity was lowered from 35 to 15 per thousand, while it decreased by 19.7 times when Cd dropped from 320 to 10 microM. "Site" did not significantly affect the mortality risk in the salinity experiment but affected time-to-death via its interaction with the "salinity-treatment". Generally, mortality did not occur at a given threshold Cd tissue level, but changed over time and treatments, in function of the site. The results demonstrate the importance of the animals' environmental history and illustrate the usefulness of time-to-death analyses in ecotoxicological experiments. PMID- 15129768 TI - Effects of water-borne 4-nonylphenol and 17beta-estradiol exposures during parr smolt transformation on growth and plasma IGF-I of Atlantic salmon (Salmo salar L.). AB - 4-Nonylphenol (4-NP) is an endocrine disrupting substance (EDS) capable of mimicking the action of 17beta-estradiol (E2). It has been hypothesized that 4-NP in a pesticide formulation is linked to historical declines in Canadian Atlantic salmon (Salmo salar L.) populations, with effects being related to exposure during parr-smolt transformation (PST). To test this hypothesis, Atlantic salmon smolts were exposed to pulse-doses of water-borne 4-NP (20 ug/l), sustained doses of water-borne E2 (100 ng/l) (positive control), or ethanol vehicle (negative control) in mid-May during the final stages of PST. Individually tagged smolts were then sampled at three times (June, July and October) to monitor subsequent growth in sea water and plasma insulin-like growth factor I (IGF-I) concentrations. Smolt weights and plasma IGF-I concentrations were both affected by E2 and 4-NP. The effects of E2 and 4-NP on mean smolt weights were most prominent in July and October [E2 (*98.1 +/- 2.8, *242.3 +/- 10.6 g), 4-NP (*102.1 +/- 3.1, 255.7 +/- 9.5 g), controls (112.5 +/- 2.8, 282.3 +/- 8.8 g)] (P < 0.05), while their effects on mean plasma IGF-I concentrations were most prominent in June and October [E2 (15.0 +/- 1.9, 28.4 +/- 1.8 ng/ml), 4-NP (*14.8 +/- 1.9, *21.6 +/- 1.7 ng/ml), controls (20.0 +/- 1.1, 31.1 +/- 2.0 ng/ml)] (P < 0.05). Additionally, results suggest that the mechanisms of action of E2 and 4-NP involve disruption in the GH/IGF-I axis, and that they may be different from each other. The effects of E2 and 4-NP on growth and plasma IGF-I concentrations observed in this study are ecologically significant because they evoke concerns for successful growth and survival of wild salmon smolts exposed to low levels of estrogenic substances that may occur from current discharges into rivers supporting sea-run salmon stocks. PMID- 15129769 TI - Differential sensitivity of three marine invertebrates to copper assessed using multiple biomarkers. AB - Understanding how biomarkers relate to each other on exposure to particular contaminants in different species is key to their widespread application in environmental management. However, few studies have systematically used multiple biomarkers in more than a single species to determine the variability of sublethal effects of a particular contaminant. In this study, three marine invertebrates, the shore crab Carcinus maenas, the common limpet Patella vulgata and the blue mussel Mytilus edulis, were exposed over 7 days in the laboratory to environmentally realistic concentrations of the priority pollutant copper. A combination of molecular, cellular and physiological biomarkers was measured in each organism to detect the toxic effects of copper. Biomarkers included lysosomal stability (neutral red retention), neurotoxicity (acetylcholinesterase activity), metabolic impairment (total haemolymph protein), physiological status (heart rate) and induction of protective metallothionein proteins. P. vulgata was the most sensitive to copper with significant effects measured in all biomarkers at concentrations of 6.1 microg Cu l(-1). In C. maenas, cellular and neurotoxic endpoints were affected significantly only at 68.1 microg Cu l(-1). Exposure to copper also induced metallothionein production in crabs. Over a 7-day exposure period, M. edulis was the most tolerant species to copper with significant effects being observed at the cellular level only at 68.1 microg Cu l(-1) . In all three species, cellular and neurotoxic pathways were more sensitive to disruption than physiological processes (protein and heart rate). Results illustrate how a suite of biomarkers applied to different sentinel species can provide a 'diagnosis of stress', whereby, effects at the molecular level can be used to interpret the level of physiological impairment of the organism. PMID- 15129770 TI - The endocrine disrupting effect of municipal effluent on the zebra mussel (Dreissena polymorpha). AB - Municipal effluents have been shown to contain a cocktail of endocrine disrupting chemicals (EDCs). The estrogenic effect of these effluents has been demonstrated on both vertebrate and invertebrate species by the feminisation of the exposed males. This effect was investigated on the freshwater zebra mussel (Dreissena polymorpha) after exposure to tertiary treated effluent from a municipal sewage treatment works (STW). Mussels were exposed to the effluent in situ for 112 days during gametogenesis (December to mid-March). Levels of vitellin (Vn)-like proteins (the major protein found in oocytes) were measured indirectly using the alkali-labile phosphate (ALP) technique and confirmed by gel electrophoresis. Significant increases (P < 0.05) in Vn-like proteins were found in both male and female mussels after exposure to the effluent, indicating that endocrine disruption (ED) had occurred. Using High-performance thin layer chromatography (HPTLC) levels of the mussels main steroid, cholesterol were found to more than double after effluent exposure. General physiological (survival, condition, etc.) and histological effects were also investigated. Histological effects observed included a large increase in interstitial tissue between the seminiferous tubules of the gonad in male mussels exposed to effluent. Effluent samples were tested for estrogenic compounds using the toxicity identification and evaluation method (TIE). A complex mixture of compounds with estrogenic activity was found with 17beta-estradiol, 17alpha-ethynlestradiol and bisphenol A accounting for the majority of the effluents estrogenic activity. Results indicate that the zebra mussel is a suitable bioindicator of endocrine disruption in freshwater environments. PMID- 15129771 TI - The interactive effects of the antifouling herbicides Irgarol 1051 and Diuron on the seagrass Zostera marina (L.). AB - The herbicides Irgarol 1051 (2-(tert-butylamino)-4-cyclopropylamino)-6 (methylthio)-1,3,5-triazine) and Diuron (3-(3',4'-dichlorophenyl)-1,1 dimethylurea) are commonly incorporated into antifouling paints to boost the efficacy of the compound towards algae. Previous investigations have identified environmental concentrations of these herbicides as being a threat to non-target organisms, such as seagrasses. Their individual toxicity has been assessed, but they can co-occur and interact, potentially increasing their toxicity and the threat posed to seagrass meadows. Chlorophyll fluorescence (Fv:Fm) and leaf specific biomass ratio (representing plant growth) were examined in Zostera marina L. after a 10-day exposure to the individual herbicides. The EC20 for each herbicide was determined and these then used in herbicide mixtures to assess their interactive effects. Irgarol 1051 was found to be more toxic than Diuron with lowest observable effect concentrations for Fv:Fm reduction of 0.5 and 1.0 +/- microg/l and 10-day EC50 values of 1.1 and 3.2 microg/l, respectively. Plants exposed to Irgarol 1051 and Diuron showed a significant reduction in growth at concentrations of 1.0 and 5.0 microg/l, respectively. When Z. marina was exposed to mixtures, the herbicides commonly interacted additively or antagonistically, and no significant further reduction in photosynthetic efficiency was found at any concentration when compared to plants exposed to the individual herbicides. However, on addition of the Diuron EC20 to varying Irgarol 1051 concentrations and the Irgarol 1051 EC20 to varying Diuron concentrations, significant reductions in Fv:Fm were noted at an earlier stage. The growth of plants exposed to Diuron plus the Irgarol 1051 EC20 were significantly reduced when compared to plants exposed to Diuron alone, but only at the lower concentrations. Growth of plants exposed to Irgarol 1051 and the Diuron EC20 showed no significant reduction when compared to the growth of plants exposed to Irgarol 1051 alone. Despite the addition of the EC20 not eliciting a further significant reduction when compared to the herbicides acting alone for most of the mixtures, the lowest observable significant effect concentration for growth and photosynthetic efficiency decreased to 0.5 microg/l for both herbicides. Irgarol 1051 and Diuron have been shown to occur together in concentrations above 0.5 microg/l, suggesting that seagrasses may be experiencing reduced photosynthetic efficiency and growth as a result. PMID- 15129772 TI - 3-Methylcholanthrene induces lymphocyte and phagocyte apoptosis in common carp (Cyprinus carpio L) in vitro. AB - Polycyclic aromatic hydrocarbons (PAH) are an important class of environmental pollutant that are known to be carcinogenic and immunotoxic. In mammals it was suggested that PAH compromise the immune system in part through the induction of programed cell death (apoptosis). In fish, no study has reported the importance of this physiological process in PAH-induced immunotoxicity. We have therefore investigated the capacity of 3-methylcholanthrene to induce lymphocyte and phagocyte apoptosis in carp. By three criteria (exposition of phosphatidylserine at the outer cell membrane, chromatin condensation and fragmentation, and decreased cell size) the data indicate that 3-methylcholanthrene (3-MC) treatment (from 20 to 200 microM) during 24 h produces apoptosis in both lymphocytes and phagocytes. In order to evaluate whether 3-MC induced apoptosis is related to the metabolic activation of 3-MC or 3-MC Ah-R binding, co-exposure experiments with 3 MC and alpha-naphtoflavone (alpha-NF), a compound that inhibits metabolic activation of 3-MC and 3-MC Ah-R binding were performed. While alpha-NF did not prevent 3-MC-induced apoptosis, the compound itself was found to be a strong inducer of apoptosis. There results might indicate that metabolic activation of 3 MC or 3-MC Ah-R binding is not causally linked to apoptosis. However, since 3-MC, alpha-NF and 3-MC + alpha-NF treatments produce the same sustained increase (3 h minimum) in intracellular calcium level, it is possible that this phenomenon is implicated in the induction of programmed cell death by these hydrocarbons. PMID- 15129773 TI - Toxicological effects of malachite green. AB - This review summarises the wide range of toxicological effects of malachite green (MG), a triarylmethane dye on various fish species and certain mammals. MG is widely used in aquaculture as a parasiticide and in food, health, textile and other industries for one or the other purposes. It controls fungal attacks, protozoan infections and some other diseases caused by helminths on a wide variety of fish and other aquatic organisms. However, the dye has generated much concern regarding its use, due to its reported toxic effects. The toxicity of this dye increases with exposure time, temperature and concentration. It has been reported to cause carcinogenesis, mutagenesis, chromosomal fractures, teratogenecity and respiratory toxicity. Histopathological effects of MG include multi-organ tissue injury. Significant alterations occur in biochemical parameters of blood in MG exposed fish. Residues of MG and its reduced form, leucomalachite green have been reported from serum, liver, kidney, muscles and other tissues as also from eggs and fry. Toxicity occurs in some mammals, including organ damage, mutagenic, carcinogenic and developmental abnormalities. However, despite the large amount of data on its toxic effects, MG is still used as a parasiticide in aquaculture and other industries. It is concluded that the potential of alternative parasiticides, like humic acid, chlorine dioxide and Pyceze, should be explored to replace MG. Until then, MG should be used with extreme care at suitable concentrations and at times when the temperature is low. Removal of residual MG in treatment ponds should also be considered. PMID- 15129774 TI - Copper detection in the Asiatic clam Corbicula fluminea: optimum valve closure response. AB - When exposed to a contaminant, bivalves close their shell as a protective strategy. The aim of the present study was to estimate the maximum expected dissolved copper sensitivity in the freshwater bivalve Corbicula fluminea using a new approach to determine their potential and limit to detect contaminants. To take into account the rate of spontaneous closures, we integrated stress problems associated with fixation by a valve in usual valvometers and the spontaneous rhythm associated with nycthemeral activity, to optimize the response in conditions where the probability of spontaneous closing was lowest. Moreover, we used an original system with impedance valvometry, using lightweight impedance electrodes, to study free-ranging animals in low stress conditions combined with an analytical approach describing dose-response curves by logistic regression, with valve closure reaction as a function of response time and concentration of contaminant. In C. fluminea, we estimated that copper concentrations > 4 microg/l (95% confidence interval (CI95%), 2.3-8.8 microg/l) must be detected within 5 h after Cu addition. Lower values could not be distinguished from background noise. The threshold values were 2.5 times lower than the values reported in the literature. PMID- 15129775 TI - The relation between latent inhibition and symptom-types in young schizophrenics. AB - Latent inhibition (LI), retarded conditioning to a stimulus that has been previously repeatedly presented without reinforcement, was examined in young schizophrenics and normal controls using a within-subject visual search task. Healthy controls exhibited the usual LI effect. LI was potentiated in schizophrenics who simultaneously exhibited high levels of negative symptoms and low levels of positive symptoms. Schizophrenic groups with other combinations of positive and negative symptoms did not differ from controls. The pattern of data suggests that past inconsistencies in the LI-schizophrenia literature may be the result of opposing processes that are associated with positive and negative symptoms. PMID- 15129776 TI - Modeling a task that is sensitive to dementia of the Alzheimer's type: individual differences in acquisition of a visuo-spatial paired-associate learning task in rhesus monkeys. AB - Early detection of progressive diseases such as Alzheimer's Disease (AD) is crucial for both the treatment and study of the disease. Performance on a visuo spatial paired-associates learning (vsPAL) task was recently shown to reliably predict a diagnosis of AD in aged populations. The present study reports the development of this vsPAL task for use in nonhuman primates. Translation of vsPAL to a nonhuman model may provide improved preclinical tools for study of the etiology and treatment of dementia. Twelve young adult male rhesus monkeys were trained to perform the vsPAL task concurrently with tests comprising a nonhuman primate neuropsychological test battery. Monkeys successfully learned to perform vsPAL and did so in a task-difficulty ranked fashion. Despite significant individual differences in capability in the acquisition of the recognition memory aspects of the task, all monkeys evidenced the ability to learn within-trial, i.e. to improve with repeated stimulus-location pairings. These results support the use of vsPAL performance under various challenge conditions to investigate the possible substrates of early cognitive decline in AD. Comparison of performance on vsPAL with performance on other memory tasks in the battery will be of more general use in differentiating mechanisms involved in various aspects of mnemonic function. PMID- 15129777 TI - Acute and long-term consequences of single MDMA administration in relation to individual anxiety levels in the rat. AB - Our previous work has shown that normal male Wistar rats can differ systematically in their behavioral response to the elevated plus-maze (EPM), where animals with high (HA) or low anxiety (LA) levels can be identified based on the percentage of time spent in the open arms. These animals also differ in other behavioral tests (e.g. active avoidance), and in their serotonin levels in the ventral striatum. Here, we tested whether such HA and LA rats might respond differently to the amphetamine analogue 3,4-methylenedioxymethamphetamine (MDMA, "ecstasy"). This drug can affect psychomotor activation and anxiety; effects which are probably due to its pronounced serotonergic and dopaminergic impacts in the rat brain. Based on a routine screening procedure in the plus-maze, male Wistar rats were divided into HA and LA sub-groups, in which rectal temperature was measured. Thirty minutes after the i.p. injection of MDMA (7.5 or 15 mg/kg) or vehicle, they were again tested in the plus-maze. During the next 3 weeks, the animals underwent further behavioral tests (plus-maze, open field, active avoidance, forced swimming) to test for possible long-term consequences of MDMA. Rectal temperature was found to be higher in LA than HA rats and was especially increased with the higher dose of MDMA (15 mg/kg). In the acute plus-maze test, the lower dose of MDMA led to an anxiogenic-like profile, whereas the higher dose led to an anxiolytic-like profile, both in HA and LA rats. Possible long-term consequences of MDMA were only tested with 7.5 mg/kg MDMA, since the 15 mg/kg dose led to a high level of lethality. The analysis of open field, plus-maze (performed after 9-12 days), and forced swimming behavior (performed after 20-21 days) did not provide indications for lasting effects of MDMA. In contrast, active avoidance learning was impaired in LA- but not HA-rats treated with MDMA. A single injection of MDMA does not only have acute effects on anxiety and psychomotor activation, but can also have some prolonged or delayed task dependent behavioral consequences. The detection of such sequels can require that individual differences are taken into account and here, determining anxiety levels in the EPM seems to serve as a useful approach. PMID- 15129778 TI - Relationship between vision and motor impairment in children with spastic cerebral palsy: new evidence from electrophysiology. AB - The aim of the present study was to measure visual acuity (VA) by the sweep visual evoked potential method (sVEP) and relate it to the degree of motor impairment in children with spastic cerebral palsy (SCP). Monocular VA was estimated in 37 SCP children aged from 6 to 48 months, classified as tetraplegic (n = 14), diplegic (n = 13), and hemiplegic (n = 10), without ophthalmological complaints with ages ranging from 6 to 48 months. Motor impairment was rated according to the Gross Motor Function Classification System (GMFCS), in five levels of severity. VA was below age norms in 13/14 (92%) tetraplegics, 10/13 (77%) diplegics and 4/10 (40%) hemiplegics. In addition, a two-way ANOVA within each subgroup showed significant differences in VA between the five GMFCS levels, with high positive correlation between VA loss and the GMFCS rating. Differences between the three types of SCP impairment in each level of GMFCS were not statistically significant, possibly due to the small number of patients. In conclusion, the use of an electrophysiological method (sweep-VEP) for the measurement of visual acuity in these patients allows a more precise and reliable estimate than behavioral measurements, since their motor impairment might interfere with the behaviorally assessed visual acuity. In addition, the finding of a high correlation between quantified motor impairment and VA loss in SCP patients is a new observation that might help to understand the causes of VA loss in these patients. PMID- 15129779 TI - Different roles of 5-HT2A and 5-HT2C receptors in regulation of female rat paced mating behaviour. AB - Serotonin (5-HT) receptor 2A (5-HT2A) and 2C (5-HT2C) agonists have been reported to facilitate female rat lordosis behaviour. This study investigated the acute effects of the 5-HT2A receptor agonist DOI ((+/-)-1-(2,5-dimethoxy-4-iodophenyl) 2-aminopropane) and the 5-HT2C receptor agonist MK-212 (2-chloro-6-(1 piperazinyl) pyrazine) on paced mating behaviour in a population of sexually receptive female rats in order to explore the role of 5-HT2A and 5-HT2C receptors in the mediation of female rat sexual motivation. DOI (0.5 and 1.0 mg/kg) increased female rat sexual motivation during the first of two consecutive copulatory series seen as a tendency towards a decrease in return latencies following ejaculation and decreased inter-intromission intervals. MK-212 generally increased approach latencies. 0.5 mg/kg MK-212 increased post ejaculatory exit latency, while 1.0 and 2.0 mg/kg MK-212 increased both post ejaculatory exit latencies and post-ejaculatory return latencies. The possibility that an increased level of anxiety confounded the effects of MK-212 on sexual motivation behaviour is discussed. The results may support the hypothesis that both 5-HT2A and 5-HT2C receptors are important in regulation of female sexual behaviour. PMID- 15129780 TI - The prenatal methylazoxymethanol acetate treatment: a neurodevelopmental animal model for schizophrenia? AB - The prenatal methylazoxymethanol acetate (MAM) treatment has been proposed as a suitable model for the neurodevelopmental aspects of schizophrenia since the morphological abnormalities it induces in the brain are subtle and in line with most reports of neuropathology in schizophrenic brains. However, the functional aspects of this treatment have not been investigated with behavioural paradigms that are relevant for the psychopathology of the symptoms of schizophrenia. In the present study, we investigated the validity of the prenatal MAM treatment as a developmental model for schizophrenia with a prepulse inhibition of the acoustic startle reflex, latent inhibition, locomotor activity, and cognition and emotionality with freezing in fear conditioning paradigms. We have conducted two studies: in Study I, MAM was injected from E09 to E12, and in Study II MAM was administered at later stages in the embryonic development, from E12 to E15. Morphologically, the prenatal MAM treatment induced mild to severe reduction in brain weights and in the entorhinal cortex, prefrontal cortex and striatum volumes, the severity of the effects depending on the timing of administration. However, despite the morphological abnormalities induced by the MAM treatments, no behavioural deficits were observed in the MAM-treated animals when compared to Controls in prepulse inhibition, latent inhibition with the two-way active avoidance, and in the freezing paradigms. Therefore, due to the consistent lack of treatment effect observed in the present investigation, we conclude that the prenatal MAM treatment has no validity as a behavioural model for schizophrenia. PMID- 15129781 TI - Response disinhibition may be explained as an extinction deficit in an animal model of attention-deficit/hyperactivity disorder (ADHD). AB - Attention-deficit/hyperactivity disorder (ADHD) is a disorder affecting between 2 and 12% of grade-school children disturbing social, academic, and occupational functioning. Problems related to social adjustment and functioning and/or psychiatric problems will exist in 50-70% of adolescents and young adults diagnosed with ADHD as children. It has been suggested that altered reinforcement and extinction processes may cause the symptoms of ADHD. The present study investigated extinction processes in spontaneously hypertensive rats (SHR), possibly the best-validated animal model of ADHD. Extinction was tested after either a variable interval (VI) or a fixed interval (FI) schedule of reinforcement with and without the presence of a conditioned reinforcer (light in the water cubicle). The results indicate a slower extinction process in the SHR compared to the normal controls, especially during the initial transition from scheduled reinforcement to extinction. Also, more responses were retained in the SHR during the later part of extinction. The extinction deficit in the SHR may be linked to reinforcer unpredictability and the presence of conditioned reinforcers, and may explain response disinhibition seen in children with ADHD. PMID- 15129782 TI - P2 receptor-mediated effects on the open field behaviour of rats in comparison with behavioural responses induced by the stimulation of dopamine D2-like and by the blockade of ionotrophic glutamate receptors. AB - The effects of the P2 receptor ligands 2-methylthio ATP (2-MeSATP; 10 pmol)--as a non-specific agonist--and pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS; 10 pmol)--as a non-selective antagonist--after bilateral intra-accumbens injection on the locomotor response were investigated in an open field situation. The P2 receptor-mediated effects on the pattern of locomotor activity were compared with the effects caused by the dopamine D2-like receptor agonist quinpirole (10 pmol) and by the combination of the N-methyl-D-aspartate (NMDA) receptor antagonist (+/-)-3-(2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP; 10 pmol) with the alpha-amino-3-hydro-5-methyl-4-isoxazolpropionic acid (AMPA) and kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 30 pmol). The intra-accumbens injection of all tested compounds elicited an increase in the locomotor activity over a test period of 20 min when compared with the controls. No statistically significant differences could be evaluated between the different drug-treated groups. However, a more detailed analysis- using further behavioural parameters such as the number of movement direction changes, the effective running time and the running speed--revealed two basically different patterns of locomotor activity. The locomotor response induced by the injection of 2-MeSATP or quinpirole was characterised by a continuous and consistent locomotion, whereas the enhanced locomotor activity elicited by PPADS or CPP/CNQX was determined by an increased running speed accompanied by more disruptions and more changes of movement direction. The coadministration of 2 MeSATP and quinpirole led to an enhancement of locomotor activity in a limited post-treatment interval. The effects of both compounds could be abolished by the pre-treatment with the D2/D3 receptor antagonist sulpiride (100 pmol). Coadministration of PPADS and CPP/CNQX caused additive effects suggesting that the pathway mediated by P2 and ionotrophic glutamate receptors is different. The stimulation of P2 receptors in the nucleus accumbens (NAc) modulates the locomotion in the direction to be to be longer lasting, more consistent and more goal directed. PMID- 15129783 TI - Corticosterone facilitates retention of contextually conditioned fear and increases CRH mRNA expression in the amygdala. AB - The present study examined the effects of glucocorticoid administration on emotional memory and on corticotropin-releasing hormone (CRH) mRNA expression in the central nucleus of the amygdala (CeA) and the paraventricular nucleus of the hypothalamus (PVN). This was tested by administering repeated corticosterone (CORT) within a contextual fear conditioning paradigm. Rats received 2.5 mg/kg (s.c.) CORT or placebo twice a day for five and a half days and, 2 h after the last injection, rats were given one-trial contextual fear conditioning. When tested for retention of conditioned fear 6 days later, the CORT-treated rats displayed more fear-conditioned freezing in the retention test than vehicle treated rats, which was not accounted for by an increase in footshock responsivity nor elevated plasma CORT. Another group of rats was fear conditioned prior to CORT administration, followed 24 h later by the five and a half days of CORT, and tested 6 days later; conditioned fear was not enhanced in these rats. Finally, CORT administration produced an increase of CRH mRNA in the CeA and a decrease in the PVN. The data suggest that repeated administration of CORT given before fear conditioning facilitates the acquisition of emotional memory, whereas CORT given after consolidation does not increase emotional memory. PMID- 15129784 TI - Contrast-induced nephropathy. PMID- 15129785 TI - Hypertriglyceridemia and coronary artery disease--an update. PMID- 15129786 TI - Determinants of left atrial pressure in rheumatic mitral stenosis: role of left atrial compliance and "atrial stiffness". AB - BACKGROUND: A wide range of left atrial pressures exist in rheumatic mitral stenosis despite similar mitral valve area. Left atrial compliance may be an important determinant of left atrial pressure in mitral stenosis. Data regarding left atrial compliance in rheumatic mitral stenosis and changes following balloon mitral valvotomy are scarce. METHODS AND RESULTS: Left atrial compliance and predictors of left atrial pressure were analyzed in 85 patients with mitral stenosis undergoing balloon mitral valvotomy. The stroke volume was divided by systolic rise in left atrial pressure to calculate the left atrial compliance. Systolic rise in left atrial pressure was computed as difference between amplitudes of left atrial "v" wave and "x" descent. The mean left atrial compliance prior to balloon mitral valvotomy was 2.62+/-1.20 cm3/mmHg. Following successful balloon mitral valvotomy there was a significant fall in pulmonary artery pressure, mean left atrial pressure, transmitral gradient, and significant increase in cardiac output, stroke volume and mitral valve area. There was a marked increase in left atrial compliance from 2.62+/-1.20 to 6.1+/-3.16 cm3/mmHg. On univariate analysis pulmonary artery systolic pressure, pulmonary artery diastolic pressure, pulmonary artery mean pressure, mean transmitral gradient, mitral valve area and left atrial compliance were the only correlates of left atrial pressures, while no correlation was noted with age, gender, left atrial size, cardiac output and stroke volume. Those with higher pulmonary artery pressure, higher transmitral gradient, lower mitral valve area and lower left atrial compliance had higher left atrial mean pressure, and the strongest negative correlation was noted with left atrial compliance. On multivariate analysis the strongest predictors of left atrial mean pressure were transmitral gradient and left atrial compliance. CONCLUSIONS: Patients with rheumatic mitral stenosis have markedly depressed left atrial compliance and hence have "stiff" left atria. Left atrial compliance is an important determinant of left atrial pressure, and improves immediately after successful balloon mitral valvotomy, irrespective of pre-balloon mitral valvotomy left atrial pressures. PMID- 15129787 TI - An echocardiographic study of net atrioventricular compliance in juvenile and adult mitral stenosis. AB - BACKGROUND: Left atrial compliance is an important determinant of symptoms in mitral stenosis. About one-third of patients with mitral stenosis have reduced left ventricular compliance. We measured the net atrioventricular compliance in rheumatic mitral stenosis patients noninvasively and analyzed if there were any clinical, electrocardiographic, roentgenographic or echocardiographic correlates of net atrioventricular compliance. METHODS AND RESULTS: Seventy-six patients with mitral stenosis were analyzed and as many normal subjects were taken as control group. Patients were divided into two groups--those 20 years and below were grouped as juvenile mitral stenosis and those above 20 years as adult mitral stenosis patients. The net atrioventricular compliance in patients with mitral stenosis was significantly impaired compared to normal population. Mean compliance in juvenile group was 4.66+/-2.18 ml/mmHg (range 2.17-9.6) and in adult group it was 4.79+/-1.99 ml/mmHg (range 2.04-8.9) (p = ns). There was no difference in net atrioventricular compliance between the juvenile and adult patients with mitral stenosis. Mitral valve area showed an independent positive correlation with net atrioventricular compliance. CONCLUSIONS: The net atrioventricular compliance was significantly reduced in patients with rheumatic mitral stenosis; however, there was essentially no difference in the net atrioventricular compliance between the juvenile and adult patients with mitral stenosis. The net atrioventricular compliance may not be responsible for the more severe symptoms observed in juvenile mitral stenosis. PMID- 15129788 TI - Minimally invasive pulmonary artery banding: a new approach. AB - BACKGROUND: A variety of approaches have been described for banding of the pulmonary artery. The indications for this procedure are limited; however in developing countries, many patients still need pulmonary artery banding for a variety of reasons. We describe a new approach, minimally invasive, using only a split in the manubrium sterni to conduct the procedure. METHODS AND RESULTS: Between January 2000 and May 2002, 19 patients who had undergone pulmonary artery banding using a minimally invasive technique were compared with 20 cases of pulmonary artery banding performed by the conventional technique. The mortality was similar in the two groups (p=0.45). The period of intubation and duration of intensive care unit stay were significantly shorter in the minimally invasive group (p=0.015 and 0.002, respectively). The duration of hospital stay was not significantly different between the 2 groups (p=0.139). In the minimally invasive group, three patients underwent subsequent reoperation. CONCLUSIONS: Minimally invasive pulmonary artery banding is useful in babies with high-flow cardiac lesions and cardiac cachexia. PMID- 15129789 TI - Redo surgery after permanent pacemaker implantation: performance indicator of surgical techniques and pacing technology. AB - BACKGROUND: The incidence of permanent pacemaker-related complications is reducing due to advancement of technology and increasing operator experience. There are only few series from India reporting the annual complication rates from a single center over the years. This is a series of 782 pacemakers implanted over 20 years in a secondary healthcare set-up. METHODS AND RESULTS: Eighty-two patients underwent redo surgery, either for a procedure-related complication (n=34) or for replacement of a malfunctioning/end-of-life pacemaker (n=48). Through critical analysis and corrective measures, all the procedural complications were reduced to less than 4% of the annual implantations. The introduction of a pacemaker follow-up clinic contributed to reducing the rate of elective replacement for battery depletion from 19.17% of the implanted VVI pacemakers to 0.63%. CONCLUSIONS: Despite being a low-volume center, with less than 100 pacemakers implanted annually, the performance of our pacing practice has shown continuous improvement. PMID- 15129790 TI - Angiotensin-converting enzyme gene polymorphism in coronary artery disease in north India. AB - BACKGROUND: The aim of this study was to investigate the role of angiotensin converting enzyme gene polymorphism in patients with coronary artery disease in north India. METHODS AND RESULTS: One hundred forty-six patients with angiographically proven atherosclerotic coronary artery disease, and 146 age- and sex-matched control subjects (treadmill-negative) were included in the study. Genomic DNA was extracted and analyzed for angiotensin-converting enzyme insertion/deletion polymorphism. Two independent investigators scored the genotypes. CONCLUSIONS: When we compared the genotypes of patients with coronary artery disease with those of normal controls, it was seen that all three genotypes, i.e. DD, ID and II, were not statistically different among patients and controls. Further, we categorized the patient and control groups into 2 subgroups, i.e. below and above 50 years of age. Interestingly, it was observed that the DD genotype was significantly higher in patients in the higher age group (i.e. above 50 years of age). However, this needs further validation by studying patients with coronary artery disease from other parts of India. PMID- 15129792 TI - Radiofrequency perforation for pulmonary atresia and intact ventricular septum. AB - Surgical or interventional perforation of atretic pulmonary valve as initial palliative procedure has been advocated in patients with pulmonary atresia with an intact ventricular septum. We report two infants with pulmonary atresia and intact ventricular septum, who were treated by radiofrequency perforation and balloon dilatation. PMID- 15129791 TI - Does device-based testing save time during automatic implantable cardioverter defibrillator implantation? AB - BACKGROUND: Defibrillation testing can be done either via an external cardiac defibrillator or directly via the implanted defibrillator during implantation (device-based testing). The advantage of one testing methodology over the other has not been adequately studied. METHODS AND RESULTS: Seventy-four patients (72% men) were randomized into two groups depending on the defibrillation testing methodology used--external cardiac defibrillation and device-based testing groups. R-wave, pacing threshold, pacing impedance, defibrillation threshold, defibrillation pathway impedance and total procedure time were not significantly different between the two groups. CONCLUSIONS: Device-based testing did not significantly reduce the procedure time. Lead and defibrillation parameters were similar in both the groups; lead repositioning and replacement were required in three patients in the external cardiac defibrillation group. PMID- 15129793 TI - Percutaneous transluminal angioplasty using a cutting balloon for stenosis of the arch vessels in aortoarteritis. AB - Percutaneous transluminal angioplasty with or without stenting of steno occlusions in the supra-aortic arch vessels in aortoarteritis is associated with suboptimal results and higher restenosis rates, particularly in those with diffuse disease. We report two cases of stand-alone cutting balloon angioplasty for patients who presented with critical stenosis of the common carotid artery, and occlusion of the subclavian artery, respectively. PMID- 15129794 TI - Pace mapping for the localization of focal atrial tachycardia arising near the mitral annulus. AB - Of the various therapeutic modalities available to treat ectopic atrial tachycardia, radiofrequency catheter ablation has shown excellent results. It is usually possible to localize the earliest site of endocardial activation by conventional or newer three-dimensional mapping techniques. We report a case of ectopic atrial tachycardia, wherein the tachycardia was being repeatedly interrupted by mechanical trauma. Finally, with the help of P wave pace mapping, the tachycardia was localized near the posterolateral part of the mitral annulus, and successfully ablated. This report demonstrates the utility of P wave pace mapping in ectopic atrial tachycardia. PMID- 15129795 TI - Isolated thrombus producing tricuspid stenosis: an unusual presentation in primary antiphospholipid syndrome. AB - We report the case of a female patient who presented with signs and symptoms of tricuspid stenosis. Echocardiography revealed a sessile mass on the tricuspid valve. A diagnosis of primary cardiac tumor was made but histopathologic examination revealed the mass to be an organized thrombus. Subsequent serological tests showed elevated titers of anticardiolipin antibodies in the absence of any associated disease. This case highlights an unusual presentation of the primary antiphospholipid syndrome. PMID- 15129796 TI - Anterior mediastinal teratoma presenting as cardiac tamponade. AB - Anterior mediastinal teratoma presenting clinically as cardiac tamponade is unusual. We report the case of a 58-year-old woman, with an unremarkable prior history, who was admitted with cardiac tamponade. Emergency pericardial aspiration showed a thick, yellow fluid. Computed tomography scan of the chest was suggestive of anterior mediastinal teratoma rupturing into the pericardium. Within 24 hours, just before the planned surgery could be performed, it re ruptured, warranting emergency surgery. Histopathology confirmed the diagnosis of a teratoma. Early surgery is recommended for even asymptomatic benign mediastinal teratomas to avoid this potentially lethal complication of rupture into the pericardial cavity, and cardiac tamponade. PMID- 15129797 TI - Evaluation of the efficacy, safety and tolerability of intravenous enalaprilat in the treatment of grade III essential hypertension in Indian patients. AB - Hypertensive emergency is a common problem requiring an effective, safe and easily administrable agent to reduce the blood pressure. Favorable data on injectable enalaprilat have been reported from the West but no Indian study has been done in such settings. We studied 10 patients (5 male, 5 female), with mean age 47+/-0.6 years and mean blood pressure 196+/-18.95/119.4+/-19.53 mmHg, who were given 1.25 mg intravenous enalaprilat. Reduction in their blood pressure started at 5 min with peak reduction noted at 4 hours (155.25+/-29.54/93.5+/ 13.55 mmHg). No adverse symptoms or biochemical changes were noted. Thus, we conclude that intravenous enalaprilat is an effective, safe and well tolerated agent for managing severe hypertension in patients requiring an emergency reduction in blood pressure. PMID- 15129798 TI - Anomalous drainage of the right superior vena cava into the left atrium. PMID- 15129799 TI - Pro-oxidant malonaldehyde and antioxidant ceruloplasmin levels in hypertrophic cardiomyopathy. PMID- 15129800 TI - Right-sided pneumothorax simulating anterior wall myocardial infarction. PMID- 15129801 TI - Platelet cGMP inversely correlates with age in healthy subjects. AB - Biochemical modifications associated with the increase in platelet activity with age are not well defined. Furthermore it is well known that the nitric oxide/cyclic 3', 5'-guanosine monophosphate (cGMP) pathway regulates platelet aggregation. The aim of the present study was to examine the relationship between platelet content of cGMP and age. 120 normal subjects, evaluating the cGMP platelet concentration, 17betaE2, IGF-I, dehydroepiandrosterone sulphate (DHEAS), insulin, plasma glucose, lipid pattern, homocysteine and PAI-I antigen, were studied. The multivariate analysis in a model with cGMP as dependent variable and with age, LDL, apolipoprotein B (ApoB), DHEAS, E2 and insulin-like growth factor (IGF)-I as independent variables shows a negative correlation between cGMP and age (p<0.01, beta=-0.388). In conclusion our data suggest that the reduced efficiency of the system constitutive nitric oxide synthase (cNOS)/guanylate cyclase represents at least one of the reasons of the increased platelet aggregability with age. PMID- 15129802 TI - Serum concentrations of adiponectin and leptin in patients with thyroid dysfunctions. AB - Thyroid dysfunction is associated with metabolic changes that affect mass and adipocyte function, as well as lipid and carbohydrate metabolism. Adipose tissue performs complex metabolic and endocrine functions. Leptin and adiponectin are two of the most important adipocytokines, both involved in the regulation of intermediate metabolism. The aim of this study was to evaluate the relationships between thyroid status and circulating levels of the two adipose tissue hormones. We studied 15 patients with hyperthyroidism, 15 patients with hypothyroidism and 15 euthyroid subjects, all matched by sex, age and body mass index (BMI). Serum concentrations of free thyroxine, free tri-iodothyronine, thyrotropin, leptin and adiponectin and anthropometric parameters (weight, height, BMI) were assessed. No significant difference was found among the 3 groups, as assessed by Student's t test, both for adiponectin and leptin. We conclude that metabolic changes associated with thyroid dysfunction are not related to variations in serum levels of adiponectin or leptin. PMID- 15129803 TI - The effects of laparascopic cholecystectomy operation on C-reactive protein, hormones, and cytokines. AB - The endocrine and immune changes associated with surgery are well documented, but the interaction between them has not been fully evaluated. We, therefore, investigated the possible effects of laparascopic cholecystectomy operation on C reactive protein, hormones, and cytokines. Thirty-five patients were enrolled in the study (26 females and 9 males, aged 42.4 +/- 16.9 yr, who were admitted to General Surgery Clinic for laparascopic cholecystectomy). Serum C-reactive protein, cortisol, thyroid stimulating hormone, free tri-iodothyronine, free thyroxine, tumor necrosis factor-alpha, interleukin-1beta, soluble interleukin-2 receptor, interleukin-6, and interleukin-8 levels were evaluated 6 h before and after the operation. Postoperative cortisol (p=0.02), TSH (p=0.034), tumor necrosis factor-alpha (p=0.003), soluble interleukin-2 receptor (p=0.004) were found to be significantly higher than their corresponding preoperative levels. However, postoperative serum free thyroxine (p=0.011), and free tri-iodothyronine levels (p=0.001) were decreased when compared with preoperative levels, respectively. No significant difference was observed in C-reactive protein, interleukin-1beta, interleukin-6, and interleukin-8 levels. C-rective protein levels did not change in the early period of the surgical stress. Cytokine responses observed in the early period were found to be conflicting. Cortisol is an important marker of surgical stress which also has close relationship with thyroid functions. PMID- 15129805 TI - Rapid decrease of leptin in middle-aged sedentary individuals after 20 minutes of vigorous exercise with early recovery after the termination of the test. AB - This study investigated the leptin response to vigorous exercise. We examined 12 sedentary subjects (7 males and 5 females) aged 45-59 yr (53 +/- 6.3) with body mass index of 26.1 +/- 8 Kg/m2. The selection of the population was based on the absence of endocrine or any other pathological anomaly. Basal concentrations of leptin, cortisol, insulin and glucose were measured at 08:00 h after an overnight fast. After that the individuals were placed on a computer-controlled ergometer performing a 20-min run at 70% VO2 max under controlled environmental conditions. Blood samples were obtained immediately after the completion of the test. For the following hour, all subjects were placed in the supine position and blood samples were taken at the end of the time period. Statistical evaluation was performed using analysis of variance (ANOVA) for independent variables. Plasma leptin levels exhibited a statistically significant decrease at the end of the 20 min running period (1.5 +/- 0.1 ng/ml vs 3.2 +/- 0.4 ng/ml, p<0.005). Interestingly, after 1 h in the supine position, leptin levels reached the basal values (3.17 +/ 0.1 ng/ml). The concentration of insulin, glucose and cortisol were unaltered during the exercise test (9.8 +/- 1.3 vs 8.85 +/- 1.27 microIU/ml, 95.58 +/- 6.71 mg/dl vs 98.4 +/- 0.78 mg/dl and 10.35 +/- 0.74 microg/dl vs 9.5 +/- 0.7 microg/dl respectively). In conclusion, our data demonstrate a relationship between stressful physical activity and plasma leptin levels in middle-aged subjects. PMID- 15129804 TI - Evaluation of a DHPLC-based assay for rapid detection of RET germline mutations in Italian patients with medullary thyroid carcinoma. AB - Causative gain-of-function mutations of the RET tyrosine-kinase receptor gene have been reported in more than 95% of inherited cases of medullary thyroid carcinoma (MTC; OMIM# 155240). Most RET activating mutations are clustered in mutational "hot spots" in exons 10, 11, 13, 14, 15 and 16 and are usually detected by single-strand conformation polymorphism (SSCP) followed by direct sequencing. To improve sensitivity, time and costs of mutational screening we have developed a denaturing high performance chromatography (DHPLC) protocol, based on the detection of heteroduplex molecules by ion-pair reverse-phase liquid chromatography under partially denaturing conditions. The mutational screening of RET exons 10, 11, 13-16 was performed in a total of 111 subjects, including 45 MTC patients and 49 relatives with known RET mutations and 17 individuals, being at risk of hereditary MTC and carrying unknown RET alleles. Heteroduplex peaks with a distinct and reproducible DHPLC elution profile allowed the detection of both rare and common RET mutations. Overall, the DHPLC-based methodology showed a high level of sensitivity and accuracy, nearing 100%. Furthermore, our protocol showed the ability to identify: 1) all the mutated codons of RET located in the "hot spots" domain; 2) the different point mutations occurring in the same codon of RET gene; 3) less frequent or rare mutations; 4) polymorphisms. As such, it can be proposed as a relatively simple and highly accurate method for a rapid genetic testing for members of MTC families. PMID- 15129806 TI - Gender-, age-, body composition- and training workload-dependent differences of GH response to a discipline-specific training session in elite athletes: a study on the field. AB - Ninety-nine Italian elite athletes (61 M, 38 F, mean age +/- SE: 24.1 +/- 0.6 yr, age range: 17-47 yr) of different disciplines volunteered to participate in this investigation. Basal GH concentrations were significantly higher (p<0.0001) in females (6.2 +/- 1.1 ng/ml) vs males (1.9 +/- 0.5 ng/ml). Basal GH values were negatively correlated with age and body mass index (BMI); no significant correlation was found between GH and IGF-I levels. Among female athletes, 8/38 had basal GH values higher than 10 ng/ml [2/8 athletes were taking oral contraceptives (OC)], while among males 6/61 had values higher than 5 ng/ml. In females, training sessions significantly increased (p<0.0001) basal GH concentrations (peak GH: 18.5 +/- 1.9 ng/ml), while in males GH responses were lower than in females (11.8 +/- 1.4 ng/ml, vs F: p<0.005). Six out of 38 female and 6/61 male athletes were considered GH hypo-responders (i.e. negative difference between peak GH and basal GH values), the large majority of them being subjects with elevated basal GH concentrations. In responsive athletes, peak GH values occurred immediately at the end of the training session both in males and in females; GH concentrations rapidly declined during recovery. No significant correlations were found between peak GH and age, body weight and BMI in either gender. GH responses were directly related (p<0.001) to the intensity of the workload during the sessions. In conclusion, the present study demonstrates that: 1) some elite athletes had increased GH concentrations before training, which were however associated with normal IGF-I levels; 2) GH peaks after a discipline specific training session were significantly higher in females than in males performing the same discipline, gender-related differences disappearing when post exercise total GH outputs (area under the curve) were compared; 3) peak GH values were directly correlated with training workload; 4) GH concentrations rapidly declined during recovery, values at the end of the post-training GH sampling being generally lower than those found in basal condition. PMID- 15129807 TI - Varicocele and sport in the adolescent age. Preliminary report on the effects of physical training. AB - The effect of physical training on the natural history of varicocele has received little attention. The aim of the present pilot study was to evaluate the prevalence of idiopathic varicocele in young athletes, in the attempt to find a correlation between the training workload and the clinical grade of varicocele. We evaluated 150 adolescents with an age of 10-16 yr (median age: 13 yr). All these subjects were athletes practicing different sport at agonistic level. One hundred and fifty non-athlete adolescents of matched age (median: 13.5 yr) were used as controls. All underwent physical examination, and if a varicocele was suspected, the diagnosis was confirmed or excluded by echo-color-Doppler examination. The young athletes were stratified into two groups according to the different time spent for training: Group 1-6 h training per week; Group 2-7 to 12 h per week. Statistical analysis was performed. A p<0.05 was considered significant. The physical examination revealed a clinical varicocele on the left side in 20 athletes. A significant positive correlation was observed between the Group 2 and the highest grade of varicocele (r2=0.9918, p=0.0041). In the adolescent group used as control we observed a varicocele in 16 cases. A positive correlation was observed between the number of athletes with varicocele and the highest grade of varicocele (r2=0.96, p=0.02). Sport training does not modify the prevalence of varicocele compared to the general population, but physical activity has to be considered as an aggravating factor in the natural history of varicocele. In countries where sport eligibility is necessary for agonistic sport practice, varicocele should be then considered as a conditioning factor. PMID- 15129808 TI - Characterization of binding between SF-1 and Sp1: predominant interaction of SF-1 with the N-terminal region of Sp1. AB - The transcription factor specificity protein 1 (Sp1) binds to GC boxes and interacts with many transcription factors to regulate gene expression. Steroidogenic factor-1 (SF-1) is an orphan nuclear receptor and plays a major role in regulation of the human steroidogenic acute regulatory (StAR) gene. We demonstrated that there is interaction between SF-1 and Sp1 on the human StAR promoter. In the present study, we examined the mechanism of the interaction between Sp1 and SF-1 on the human StAR gene promoter. Results of glutathione S transferase (GST) pull-down assays and a mammalian two-hybrid assay showed that SF-1 interacted with Sp1 through the N-terminal domains of Sp1. Results of electrophoretic mobility shift assays using nuclear extracts showed that Sp1 is associated with SF-1-DNA complex formation. The density of SF-1-DNA complex was much greater when recombinant Sp1 was added to the incubation mixture. These results suggest that Sp1 interacts with SF-1 and that Sp1 enhances SF-1-DNA complex formation to regulate human StAR transcription. PMID- 15129809 TI - IGF-I stimulates proliferation of spontaneously immortalized human keratinocytes (HACAT) by autocrine/paracrine mechanisms. AB - HaCaT keratinocytes are derived from adult human skin and although spontaneously immortalized, remain highly related to their normal counterparts. We observed that HaCaT cells can proliferate in serum-free medium (SFM), in contrast to normal human keratinocytes whose growth in vitro requires a feeder layer and/or the supplementation with hormones and growth factors. Since autocrine production of growth factors has been proposed as the pathway that cells may exploit to escape growth regulation, we have investigated whether this is occurring in HaCaT cultured in SFM. Either epidermal growth factor (EGF) or insulin-like growth factor-1 (IGF-I) was effective and dose-dependently stimulated HaCaT replication. The ability of these keratinocytes to express EGF and IGF-I and their receptors was investigated by northern blot and reverse transcriptase-polymerase chain reaction (RT-PCR). We report that HaCaT cells synthesize mRNAs for IGF-I, IGF-II, IGF-IR and EGF-R but not EGF mRNA. Immunoneutralization of IGF-I with specific monoclonal antibodies blocked spontaneous HaCaT proliferation in SFM, as did incubation with antibodies against IGF-IR. These data demonstrate that an autocrine/paracrine loop based on IGF-I may allow HaCaT keratinocytes to proliferate autonomously in culture in contrast to keratinocytes in primary culture. A similar mechanism may be involved in the development of hyperproliferative diseases of human skin and its functional disruption may represent the target for therapeutic approaches. PMID- 15129810 TI - Snoring, hypertension and Type 2 diabetes in obesity. Protection by physical activity. AB - Sleep-related breathing disorders are recognized as major health problems in obesity. They are involved in both hypertension and Type 2 diabetes, through mechanisms possibly related to increased sympathetic tone. We studied the association of habitual snoring with diabetes, hypertension, weight cycling and physical activity in a large Italian database of treatment-seeking obese subjects. Clinical and behavioral data were assessed by standardized questionnaires. Consecutive data of 1890 obese patients were analyzed [average body mass index (BMI), 38.2 kg/m2, median age: 46 yr, 78% females], from 25 obesity Italian centers, with low prevalence of clinical manifestations of cardiovascular disease. Habitual snoring was reported in 56% of the cases, and was associated with day-time sleepiness. The prevalence increased with obesity class and waist circumference, and was positively associated with weight cycling and weight gain since the age of 20, and smoking. Regular physical activity had a protective effect. Snoring was associated with diabetes and hypertension at univariate analysis, but in multivariate analysis an independent effect was only observed for hypertension. After adjustment for age, gender and BMI, physical activity maintained an independent, protective effect on both snoring (odds ratio 0.65, 95% confidence interval 0.49-0.84; p=0.001), diabetes (0.50, 0.30-0.86; p=0.011) and hypertension (0.71, 0.53-0.95; p=0.023). We conclude that in treatment-seeking, obese subjects with low prevalence of cardiovascular disease, snoring independently increases the risk of hypertension, whereas physical activity exerts a protection on both snoring and complications. These data underline the importance of lifestyle interventions to limit the burden of obesity and associated diseases. PMID- 15129811 TI - Analysis of the vitamin D receptor Fokl polymorphism. AB - In the present study we analysed the vitamin D receptor (VDR) Fokl polymorphism distribution in a Caucasian population from the North-West of Spain. This polymorphism has been associated with variation in bone mineral density levels and has been proposed as a factor for the genetic predisposition to osteoporosis. This study was performed in 60 healthy individuals by restriction fragment length polymorphism (RFLP) analysis using the endonuclease Fokl and alternatively using single strand conformation polymorphism (SSCP). The genotype distribution was: 46.7% FF homozygotes, 43.3% Ff heterozygotes, and 10% ff homozygotes. These values do not reveal significant differences in comparison with other available Caucasoid populations, but show clear differences with respect to African American populations. The analysis of this polymorphism by SSCP reveals the existence of a novel single nucleotide polymorphism (AAT/AAC) located within the DNA binding domain of the VDR gene at position-19 codon. Thus, using a novel 109 bp hVDR gene fragment which excludes the AAT/AAC variant in the vitamin D receptor Fokl allows an easier determination by SSCP analysis of the Fokl polymorphism. PMID- 15129812 TI - Graves' ophthalmopathy and atrophic thyroiditis: a case report. AB - Graves' ophthalmopathy (GO)--also known as thyroid-associated orbitopathy or ophthalmopathy--usually affects patients with Graves' disease. Antibodies stimulating the TSH receptor are thought to be involved in the pathogenesis of this important and disabling extra-thyroidal manifestation of Graves' disease. Less frequently, GO occurs in subjects who neither have nor have ever shown evidence of thyroid dysfunction ("euthyroid GO"), while the occurrence of GO in patients with autoimmune Hashimoto's thyroiditis is thought to be quite rare and has sporadically been reported. The late and abrupt occurrence of severe GO without hyperthyroidism in an 88-yr-old woman with primary myxedema due to atrophic thyroiditis must be considered as an exceptional event. In this patient, GO was combined with elevated titres of serum auto-antibodies directed against the TSH receptor, while serum levels of anti-thyroglobulin and thyroperoxidase antibodies were within the normal range or only occasionally slightly above the normal values. PMID- 15129813 TI - A novel deletion mutation in the arginine vasopressin receptor 2 gene and skewed X chromosome inactivation in a female patient with congenital nephrogenic diabetes insipidus. AB - X-linked nephrogenic diabetes insipidus (NDI) is a rare inherited disorder caused by mutations in the arginine vasopressin receptor 2 (V2R) gene. The clinical phenotype is fully expressed in hemizygous male patients and is usually asymptomatic in heterozygous females. In the present study, a 51-yr-old Japanese female with congenital NDI and her family members were examined. The patient developed severe hypernatremia accompanied by hypoosmotic polyuria after gynecological surgery, and was unable to concentrate urinary osmolality in response to exogenous vasopressin. Direct sequencing analysis of the propositus and her two affected sons revealed a two-nucleotide deletion change at codon 30 (g.452-453delAC) in the V2R gene, resulting in a frameshift and premature termination in translation at codon 190. The X chromosome inactivation pattern was investigated in the propositus using methylation analysis of the polymorphic CAG repeat in the androgen receptor gene, and the value for relative X chromosome inactivation of one allele was 70.2%. In conclusion, we identified a novel V2R gene mutation in a female patient and her sons with congenital NDI, and her phenotype may be caused by skewed X chromosome inactivation. PMID- 15129814 TI - "Cushing's disease of the omentum"--fact or fiction? PMID- 15129815 TI - QT interval, cardiovascular risk factors and risk of death in diabetes. AB - A prolonged QT interval is considered an indicator of increased risk of malignant ventricular arrhythmias and/or sudden death. It has been proposed that autonomic neuropathy in diabetes is related to QT interval prolongation and increased mortality rates. Several studies in Type 1 and Type 2 diabetic patients have confirmed the independent relation between prolonged QT interval duration or increased QT interval dispersion and chronic ischemic heart disease. It has been consistently shown that autonomic neuropathy is related to QT interval duration while more controversies exist on the association with QT interval dispersion. In recent years, studies have confirmed the value of QT interval as a predictor of total mortality in both diabetic and non-diabetic subjects. Moreover, several studies have shown a significant relation between QT interval prolongation and cardiovascular disease risk factors. QT interval could be used to stratify the cardiovascular risk in diabetic patients. We still do not know why QT interval is prolonged and how this abnormality leads to death. Nevertheless, QT interval is a simple, low-cost measure, easily obtainable without the need of the patient's compliance and which could help to select patients who need second level diagnostic procedures and strict observation. PMID- 15129816 TI - New paradigms in neuroendocrinology: relationships between obesity, systemic inflammation and the neuroendocrine system. AB - Obesity may be an independent risk factor for coronary artery disease and contribute to a chronic state of systemic inflammation leading to atherosclerosis and metabolic abnormalities, such as diabetes, insulin resistance, dyslipidemia and hypertension. Visceral fat, in fact, may act as an endocrine organ, synthesizing and releasing atherogenic inflammatory cytokines, whose circulating levels depend on the individual's nutritional state, and the extent and anatomical location of fat stores. Unsuspected viral infections might also be involved in enhancing autocrine/paracrine mechanisms of cytokine release from omental fat. Elevated levels of blood cytokines may interact with the neuroendocrine system, autonomic nerves and peripheral lymphatic organs. This may lead to local inflammatory reactions in many body compartments, in particular in the heart tissue, possibly affecting the process of circulatory recovery in obese subjects, and predisposing these patients to a greater risk of myocardial inflammatory disease than individuals with normal body mass index. Circulating levels of inflammatory cytokines might be considered to determine risk categories for development of cardiovascular complications in obese subjects. In addition, their reduction with pharmacological antagonists might prevent and/or control acute cardiovascular events and increase energy expenditure in obese patients, especially after surgical treatment, through reduction of cytokine inhibition of the hypothalamic-pituitary-thyroid axis. PMID- 15129817 TI - The empty follicle syndrome. AB - The empty follicle syndrome (EFS) is characterized by the lack of retrieved oocytes from follicles after ovulation induction and apparently normal follicular development for in vitro fertilization, despite repeated aspiration and flushing. The underlying mechanism of the EFS remains hypothetical. Some Authors have suggested that it is related to the "cause" leading to female infertility, whereas others have pointed to the alternative suggestion that it might reflect dysfunctional folliculogenesis, with early oocyte atresia and apparently normal hormonal response. Moreover, some Authors believe that the EFS does not exist, and that the oocyte retrieval failure is a pharmacological fault. The risk of recurrence is higher as the age of the patients increases. The EFS cannot be predicted by the pattern of ovarian response to stimulation either sonographically or hormonally. Consequently, the diagnosis of EFS is retrospective. Whatever the underlying cause of an EFS cycle, patients with an EFS cycle should be counselled regarding the possibility of recurrence of such an event in future cycles. PMID- 15129819 TI - Fistulectomy may not be the first choice treatment in a child with recurrent suppurative thyroiditis. PMID- 15129818 TI - Erectile dysfunction: expectations beyond phosphodiesterase type 5 inhibition. AB - In the last few years the pathophysiological mechanisms of erection have been partially clarified, and the molecular machinery of the cellular components of the corpus cavernosum (CC) has been widely investigated. Since erection is a vascular event and the penis is a vascular organ, there must be an intact endothelium for an erection to occur. The regulation of penile tumescence inside the CC involves a balance between contracting and relaxing factors which regulate the functional state of smooth muscle cells. Recent studies have highlighted the importance of new local factors (i.e. phosphodiesterases, rho-kinases and endothelins), and pharmacological agents are available in the armamentarium of the specialist which are targeted to modulate the function of those mediators of erection. It is now well understood that male erectile dysfunction (ED) is a symptom rather than a disease; for this reason in the near future both general practitioners and specialists in internal medicine would have to interplay with sexual medicine. This review is intended to give the clinician some basic concepts of the pathophysiology of erection with relevance to the clinical practice, and to discuss the newest therapeutic approaches for those patients who do not respond to the treatment with oral inhibitors of phosphodiesterase Type 5. PMID- 15129820 TI - Prolactin is an amyloid-related protein. PMID- 15129821 TI - The biological clock: the bodyguard of temporal homeostasis. AB - In order for any organism to function properly, it is crucial that it be table to control the timing of its biological functions. An internal biological clock, located, in mammals, in the suprachiasmatic nucleus of the hypothalamus (SCN), therefore carefully guards this temporal homeostasis by delivering its message of time throughout the body. In view of the large variety of body functions (behavioral, physiological, and endocrine) as well as the large variety in their preferred time of main activity along the light:dark cycle, it seems logical to envision different means of time distribution by the SCN. In the present review, we propose that even though it presents a unimodal circadian rhythm of general electrical and metabolic activity, the SCN seems to use several sorts of output connections that are active at different times along the light:dark cycle to control the rhythmic expression of different body functions. Although the SCN is suggested to use diffusion of synchronizing factors in the rhythmic control of behavioral functions, it also needs neuronal connections for the control of endocrine functions. The distribution of the time-of-day message to neuroendocrine systems is either directly onto endocrine neurons or via intermediate neurons located in specific SCN targets. In addition, the SCN uses its connections with the autonomic nervous system for spreading its time-of-day message, either by setting the sensitivity of endocrine glands (i.e., thyroid, adrenal, ovary) or by directly controlling an endocrine output (i.e., melatonin synthesis). Moreover, the SCN seems to use different neurotransmitters released at different times along the light:dark cycle for each of the different connection types presented. Clearly, the temporal homeostasis of endocrine functions results from a diverse set of biological clock outputs. PMID- 15129822 TI - Extracellular pH is under circadian control in Gonyaulax polyedra and forms a metabolic feedback loop. AB - This study investigates the relationship between the circadian clock and metabolism based on recordings of the extracellular pH in cultures of the marine dinoflagellate, Gonyaulax polyedra. In light-dark cycles, pH of the medium rises during the light phase and declines in the dark. The amplitude of this pH-rhythm correlates with light intensity, indicating photosynthesis (and respiration) as the driving force. The recorded extracellular pH changes probably reflect the need to control intracellular pH in spite of pH-modifying reactions. The daily pH changes are under control of the circadian clock because they continue to oscillate with a circa-24 h period in constant light, albeit with a smaller amplitude. Similar to other circadian output rhythms, the pH rhythm depends (amplitude and phase) on nitrate levels in the medium. Both the bioluminescence and the pH rhythm can also be shifted by extracellular pH-changes although Gonyaulax is rarely exposed to significant pH changes in its marine ecosystems (except for highly dense algal blooms). Because intracellular proton levels are both affecting circadian input and output they form a feedback loop with the Gonyaulax circadian system indicating complex interactions between metabolism and the circadian clock. PMID- 15129823 TI - Glycogen synthase kinase 3beta as a likely target for the action of lithium on circadian clocks. AB - Although lithium is one of the most commonly used drugs in the prophylaxis and treatment of bipolar disorder, the mechanisms underlying its therapeutic action are still unclear. Together with its mood-stabilizing effects, lithium is also known to influence the circadian clocks of several organisms including man. Circadian rhythms are altered in patients with bipolar disorder, and it is believed that these rhythms may play an important role in disease mechanisms. It is therefore possible that some of the therapeutic actions of lithium may be related to its effect on circadian clocks. Identifying the targets for lithium's action on circadian clocks would therefore be important both for understanding the mechanisms of its therapeutic effect and also in further understanding disease mechanisms in bipolar disorders. Using Drosophila melanogaster as a model system, we show that long-term administration of lithium results in lengthening of the free-running period (tau) of circadian locomotor activity rhythm of flies in constant darkness (DD). This effect occurs at concentrations similar to the plasma levels of lithium used in the treatment of bipolar disorder. The lithium treated flies also show reduced activity of one of the previously reported targets of lithium action, Glycogen Synthase Kinase 3beta (GSK 3beta). GSK 3beta has been shown to be involved in the regulation of circadian clocks as the down regulation of this protein results in an elongation of tau. The tau elongation resembles the effect seen with lithium administration in a number of organisms including man, and taken together with the earlier observations our results suggest that lithium inhibits the activity of GSK 3beta to produce its effect on circadian clocks. PMID- 15129824 TI - Seasonal variation in daily activity patterns of free-ranging European ground squirrels (Spermophilus citellus). AB - Daily aboveground activity of European ground squirrels (Spermophilus citellus) in their natural habitat was recorded with a visual scanning procedure during the active seasons of 1992 and 1993. Activity patterns were analyzed with respect to time of year and to the animal's reproductive state. Aboveground activity started on average 3.9 h (SD 0.6 h, n = 37 days) after civil twilight at dawn and ended on average 3.2 h (SD 0.9 h, n = 37 days) before civil twilight at dusk. Between onset and offset of activity, 54% was spent aboveground, of which 73% was spent foraging. Activity patterns were influenced by photoperiod, rainfall, and by reproductive state. During mating, reproductively active males started activity earlier than females and reproductively inactive males. For females, time spent foraging was high during lactation. The midpoint of daily activity was at 12:16 h (SD 0.37 h, n = 37 days). Activity patterns of European ground squirrels thus appear robustly positioned in the middle of the photoperiod. PMID- 15129825 TI - Annual pattern of human conception in the State of Texas. AB - A total of 2,828,068 State of Texas singleton conceptions, estimated from the date of the first day of the last menstrual period, were assessed for annual periodicity by multiple-component (1-yr fundamental plus the 6- and 3-month harmonics) Cosinor analysis. An annual pattern (p < 0.001) of conception was detected with a December peak, end of July-beginning of August trough and modest amplitude (total peak-to-trough variation) equal to 13.3% of the yearly mean. Annual patterns (p < 0.005) of generally comparable peak and trough times were also documented for conceptions categorized by maternal marital status, age, ethnicity, years of education, birthplace, and county (Texas-Mexico border vs. non-border) of residence. The amplitude, a measure of the prominence of the annual pattern, varied by race (two-fold greater for non-Hispanic Blacks and Hispanics than non-Hispanic Whites), age (50-60% greater for the 19-year-old and younger group than the 20-29 and 30-44-year-old groups) and maternal education (two-fold greater for those with 0-8 than 13 or more years of schooling). Annual patterns (p < 0.001) in conceptions, generally with December peak and end of July beginning of August trough, were also detected for neonatal variables categorized by birth weight, birth order, gestational age, and gender. The amplitude of the annual pattern in conceptions varied directly with gestational age and inversely with birth weight class. PMID- 15129826 TI - Actigraphic recordings of activity-rest rhythms of neonates born by different delivery modes. AB - Activity-rest behavior of 20 neonates born vaginally, 18 neonates born by medically planned Cesarean section (C-section), and 19 neonates born by medically required C-section after labor onset (all born in the thirty-seventh to forty second week of gestation) was monitored for six successive days starting in the first week of life. Actigraphy was used to record and show time patterns of activity and rest in neonates by using small wristwatch-like Actiwatch actometers. Nursing/feeding times were recorded by using the actometers' integrated event marker button. Recordings in both C-section groups were performed in the hospital; for neonates born vaginally and for some born by C section, recordings were carried out in the hospital and in their homes. In addition to the actigraphic recordings, a standardized diary was kept regularly. To assess periodic characteristics, frequency components of activity-rest behavior were analyzed using fast Fourier transformation. Amount of sleep time during daytime, nighttime, and 24 h, as well as sleep bouts during the daytime and nighttime, were compared. The majority of vaginally born neonates showed a distinct circadian frequency in their spectra. In contrast, both groups of neonates born by C-section showed significantly less distinct circadian frequencies in their spectra. All three groups showed a significant difference in amount of nighttime sleep vs. daytime sleep, with more sleep at nighttime. There were no differences in the amount of nighttime sleep, daytime sleep, and sleep time during 24 h between the groups born by different delivery modes. PMID- 15129827 TI - Circadian variation in cardiac autonomic activity: reactivity measurements to different types of stressors. AB - The role of endogenous circadian rhythmicity in autonomic cardiac reactivity to different stressors was investigated. A constant routine protocol was used with repeated exposure to a dual task and a cold pressor test. The 29 subjects were randomly divided into two groups in order to manipulate prior wakefulness. Group 1 started at 09:00 h immediately after a monitored sleep period, whereas group 2 started 12 h later. Measures of interbeat intervals (IBI), respiratory sinus arrythmia (RSA, a measure of parasympathetic activity), pre-ejection period (PEP, a measure of sympathetic activity), as well as core body temperature (CBT) were recorded continuously. Multilevel regression analyses (across-subjects) revealed significant (mainly 24 h) sinusoidal circadian variation in the response to both stressors for IBI and RSA, but not for PEP. Individual 24 + 12 h cosine fits demonstrated a relatively large interindividual variation of the phases of the IBI and RSA rhythms, as compared to that of the CBT rhythm. Sinusoidal by group interactions were found for IBI and PEP, but not for RSA. These findings were interpreted as an indication for endogenous circadian and exogenous parasympathetic (vagal) modulation of cardiac reactivity, while sympathetic reactivity is relatively unaffected by the endogenous circadian drive and mainly influenced by exogenous factors. PMID- 15129828 TI - Influence of circadian time structure on acute hormonal responses to a single bout of heavy-resistance exercise in weight-trained men. AB - Both testosterone (T) and cortisol (C) exhibit circadian rhythmicity being highest in the morning and lowest in the evening. T is a potent stimulator of protein synthesis and may possess anti-catabolic properties within skeletal muscle, and C affects protein turnover, thereby altering the balance between hormone-mediated anabolic and catabolic activity. Physiological reactions of these hormones and training adaptations may influence the post-exercise recovery phase by modulating anabolic and catabolic processes, therefore affecting metabolic equilibrium, and may lead to intensification of catabolic processes. We investigated the effect of the circadian system on the T and C response of weight trained men to heavy resistance exercise. Thirteen young (21.8 +/- 2.2 yr) weight trained men (12 months training experience) performed an eight-station heavy resistance exercise protocol on two separate occasions (AM: 06:00 h and PM: 18:00 h), completing 3 sets of 8-10 repetitions at 75% of each subject's one-repetition maximum (1-RM). Blood samples were obtained prior to, during, and following the exercise bout, and serum total T and C concentrations were determined by competitive immunoassay technique. Performing the single bout of heavy-resistance exercise in the PM as compared to the AM positively altered the C and T/C ratio hormonal response. Pre-exercise C concentrations were significantly lower (p < 0.05) in the PM session, which resulted in a lower peak value, and the accompanying increased T/C ratio suggested a reduced catabolic environment. These data demonstrate that the exercise-induced hormonal profile can be influenced by the circadian time structure toward a profile more favorable for anabolism, therefore optimizing skeletal muscle hypertrophic adaptations associated with resistance exercise. PMID- 15129829 TI - Circadian time-qualified tolerance intervals for ambulatory blood pressure monitoring in the diagnosis of hypertension. AB - The large-amplitude circadian pattern in blood pressure of healthy subjects of both genders suggests that the constant threshold currently used to diagnose hypertension should be replaced by a time-specified reference limit reflecting the mostly predictable blood pressure variability during the 24 h. Accordingly, we derived circadian time-specified reference standards for blood pressure as a function of gender. We studied 743 normotensive Caucasian volunteers (400 men and 343 women), 45.7 +/- 16.5 (mean +/- SD) years of age. Blood pressure was measured by ambulatory monitoring at 20-min intervals during the day and at 30-min intervals at night for 48 consecutive hours. Data from each blood pressure series were synchronized according to the rest-activity cycle of each individual in order to avoid differences among subjects in actual times of daily activity. Data were then used to compute 90% circadian tolerance intervals for each gender separately. The method, derived on the basis of bootstrap techniques, does not need to assume normality or symmetry in the data and, therefore, it is highly appropriate to describe the circadian pattern of blood pressure variability. Results reflect expected changes in the tolerance limits as a function of gender and circadian sampling time, as well as upper blood pressure limits below the thresholds currently used for diagnosing hypertension, especially for women. The use of these time-dependent tolerance limits for the computation of a hyperbaric index as a measure of blood pressure excess has already been shown to provide a reproducible and high-sensitivity test for the diagnosis of hypertension, which can also be used to evaluate treatment efficacy. PMID- 15129830 TI - Ethical principles and standards for the conduct of human and animal biological rhythm research. AB - Most research papers published in Chronobiology International report the findings of investigations conducted on laboratory animals and human beings. The Journal, its editors and the publication committee endorse the compliance of investigators to the principles of the Declaration of Helsinki of the World Medical Association relating to the conduct of ethical research on human beings and the Guide for the Care and Use of Laboratory Animals of the Institute for Laboratory Animal Research of the National Research Council relating to the conduct of ethical research on laboratory and other animals. Chronobiology International requires that submitted manuscripts reporting the findings of human and animal research conform to the respective policy and mandates of the Declaration of Helsinki and the Guide for the Care and Use of Laboratory Animals. The peer review of manuscripts will thus include judgment of whether or not the involved research methods conform to the standards of good research practice. This article outlines the basic expectations for the methods of human and animal biological rhythm research, both from the perspective of the fundamental criteria necessary for quality chronobiology investigation and from the perspective of humane and ethical research on human beings and animals. PMID- 15129831 TI - Diurnal variation of pain perception in young volunteers using the tourniquet pain model. PMID- 15129832 TI - XVIth international symposium on night and shiftwork. Santos, Brazil, 17-21 November 2003. New fresh facts. PMID- 15129833 TI - The perceived health status of people with psychologically derived non-epileptic attack disorder and epilepsy: a comparative study. AB - While a wealth of literature describes the short and longer term impact of living with epilepsy, there is, in contrast, very little information about the impact of psychologically derived non-epileptic attack disorder (PNEAD). In the absence of any physical disease, some clinicians may assume that this group requires little help. However, evidence suggests that there maybe a significant impact. Therefore, this study has set out to document the effects on perceived health status of PNEAD. We compared 97 people with PNEAD with a sample, case matched for age and gender, of 97 people who had been previously diagnosed with epilepsy. A questionnaire was administered to both groups requesting information about their perceived overall health (SF-36) and their clinical and demographic status. PNEAD patients described a significantly poorer profile of perceived health compared with the epilepsy group. People with PNEAD have substantially impaired perceived health status, to the extent of feeling in poorer health than people with organically explained epilepsy. Clinical interventions are necessary that can improve their perceived health. PMID- 15129834 TI - Enterocolitis: an adverse event in refractory epilepsy patients treated with levetiracetam? AB - INTRODUCTION: Levetiracetam (LEV) is a recently marketed novel anti-epileptic drug with a promising efficacy and safety profile. In this report we describe two patients who presented with enterocolitis and discuss the possible relationship with concurrent LEV intake. PATIENTS: In two patients. LEV was initiated to control refractory complex partial seizures (CPS). The first patient was treated with 1500 mg/day and complained of abdominal pain and weight loss 6 months later. Internal examination and colonoscopy revealed a punctate colitis. The second patient presented with bloody stool 1 month after LEV initiation. Colonoscopy showed punctate colitis. In both patients gastrointestinal symptoms disappeared following tapering of LEV. DISCUSSION: There are no reports in the literature describing colitis related to LEV intake. Three possible mechanisms of action are discussed. Colitis may be part of a hypersensitivity syndrome caused by LEV. Pharmacodynamic interactions with other anti-epileptic drugs, for example, carbamazepine may play a role. A haematological adverse event is another possibility since piracetam, a related molecule, has a known impact on erythrocytes and platelets. CONCLUSION: The close temporal relationship between initiation of LEV intake, symptomatic colitis and clinical improvement following LEV tapering, suggests that colitis may be a possible and previously undescribed adverse effect of LEV. PMID- 15129835 TI - Implementing a bioassay to screen molecules for antiepileptogenic activity: chronic pilocarpine versus subdudral haematoma models. AB - BACKGROUND: There is a need to discover novel chemical compounds that will inhibit the pathological process of epileptogenesis (i.e. agents that will prevent the long-term formation of an active seizure focus following a brain insult). The goal of this paper is to identify a bioassay of value in drug design when screening new chemical entities as putative antiepileptogenic agents. METHODS: We focused on two models: the pilocarpine chronic seizure model of spontaneous recurrent seizures (SRSs) and a chronic subdural haematoma model of SRSs. Both models were evaluated using more than 20 Sprague-Dawley rats for each model. RESULTS: In the pilocarpine-induced model of SRSs, 80% of animals went on to develop SRSs when the dose of pilocarpine was 380 mg/kg i.p. In 50 animals that developed SRSs, the average number of seizures per 15 days of observation was 3.8 seizures with a range of 2-23 seizures per 15-day period. The chronic subdural model was inefficient in producing SRSs. CONCLUSIONS: A pilocarpine induced SRS model of epilepsy affords a reliable model of epileptogenesis suitable for evaluating new chemical entities as putative antiepileptogenics. PMID- 15129836 TI - The role of the clinical nurse specialist in epilepsy. A national survey. AB - PURPOSE: To review and describe the key roles of the UK clinical nurse specialist in epilepsy (CNSE), and to identify the specialist nurses' contribution to care through an exploration of CNSE's perceptions of their roles. METHOD: Using the Delphi technique [Applied Project Design and Analysis, 3rd ed., Churchill Livingstone, London, 2000, p. 243] a national survey of all known UK CNSEs was completed. One hundred and thirty questionnaires identifying nine key hypotheses central to the role of the CNSE were distributed and 76 valid questionnaires returned. RESULTS: The response rate was 63% and was geographically representative of the UK population of CNSEs. CNSEs were employed in a range of hospital and community settings with differing patient groups. Seventy-two percent of respondents held higher academic nursing qualifications but only 36% had previous epilepsy or neurology experience. Thirty percent of respondents had been employed in the role of CNSE for more than 5 years and 84% were employed as a G or H grade nurse. Only 39% of CNSEs held nurse-led clinics and of those 32% were responsible for all decisions made during their clinic. Furthermore, 40% of CNSEs saw new patients who had not previously been reviewed by one of the medical team. The level of responsibility for drug management was mainly at a monitoring and advisory level but a small number of CNSEs held much greater responsibility. The responses to the nine hypotheses were compared using cross tabulations. CONCLUSION: The findings of the study and the review of the CNSE in the UK revealed that the key roles of the CNSE were difficult to define. Yet, the respondents identified that there were common core features central to their contribution to care as specialist nurses. PMID- 15129837 TI - Epilepsy--improvement of giving the diagnosis between the demands for standardisation versus individualisation. AB - Starting from Cunningham et al.'s [Seizure 11 (2002) 500] attempt to develop a guideline for giving the diagnosis of childhood epilepsy, the paper discusses the specific difficulties emerging on the way towards a standardisation and development of guidelines for the disclosure of diagnosis. The major objective of disclosure is to enhance positive adaptation towards epilepsy and its associated stressors and treatment demands. Adaptation to a chronic disease, however, depends on subjective processes of stress appraisal and coping response. Supporting adaptation by favourable strategies of disclosure therefore requires to explore and respond to the very personal perception of the medical and psychosocial consequences of the disorder. The broad interindividual variation of subjective anxieties therefore entails the necessity to individualise the procedure of telling the diagnosis in order to maximise its goodness of fit to patient and family characteristics. A procedure is suggested that integrates the individualisation of information provision and counselling, on the one side, and the efforts of standardisation and guideline development, on the other side, in order to improve resulting disclosure practice. PMID- 15129838 TI - Risk of seizure recurrence after a first unprovoked seizure: a prospective study among Jordanian children. AB - PURPOSE: There is wide variation in the reported recurrence rate after a first unprovoked seizure in children. We investigated the risk of recurrence after a first unprovoked seizure in Jordanian children and the risk factors associated with increased recurrence rate. METHODS: All consecutive patients aged 3 months 14 years who presented with their first unprovoked seizures between January 1997 and 2000, were included in a prospective study and followed up for 3 years for possible recurrence. Of the patients studied, there was slight male predominance (56.6%) and 55% of them were 2-9 years of age. Generalised seizures were reported in 75% and the remaining 25% had partial seizures. The duration of seizure was 1 4 minutes in 59%. Family history of epilepsy was positive in 31% and parental consanguinity in 32%. The role of these factors in increasing the risk of recurrence was also investigated. RESULTS: Two hundred sixty-five patients were included in the study and continued follow up for 3 years. Ninety-eight (37%) of them experienced seizure recurrence. Among the predictor factors for recurrence, partial seizure (P = 0.003) and positive family history (P = 0.000) were associated with a statistically significant increased risk. Sex, age, duration of seizure and consanguinity were not associated with increased risk of recurrence. CONCLUSION: Thirty-seven percent of the children studied experienced a second attack after a first unprovoked seizure over the 3 years follows up period. The risk of recurrence was significantly higher in children with a partial seizure (55%) and among those with a positive family history of epilepsy (59%). Age at first seizure, sex, duration of seizure and consanguinity were not significantly related to the risk of recurrence. PMID- 15129839 TI - Is omega-3 fatty acid deficiency a factor contributing to refractory seizures and SUDEP? A hypothesis. AB - Epilepsy, the commonest serious neurological condition, is associated with an increased risk in premature deaths, including an estimated 500 sudden unexpected deaths (SUDEP) per year in the UK. In some patients seizures are associated with cardiac arrhythmias, which are thought to be a major factor in SUDEP. Omega-3 fatty acids have been shown to reduce cardiac arrhythmias in animal studies and to reduce sudden cardiac deaths, thought to be due to cardiac arrhythmias, in both healthy subjects and in those who have had one myocardial infarction. Additionally, omega-3 fatty acids in animal studies and in a small clinical observation study have shown anti-seizure effects. Omega-3 fatty acid supplementation in patients with refractory seizures may reduce seizures and seizure associated cardiac arrhythmias and hence SUDEP. PMID- 15129840 TI - Seizures as the presenting symptom of brain tumours in children. AB - Seizures were the presenting clinical symptom in 10 (12%) of 81 consecutive children with a primary brain tumour treated in a tertiary paediatric oncology unit over 5 years. Nine patients experienced partial seizures, and in seven a waking electroencephalogram showed focal or lateralising abnormalities. Astrocytoma was the most common tumour histology. The delay in tumour diagnosis from the onset of seizures ranged from 2 weeks to 2 years with a mean of 6 months. Complete resection of the tumour was the only treatment in three patients and four underwent resection followed by radiotherapy and/or chemotherapy. Two patients died. Three patients became seizure free receiving no antiepileptic medication and the remaining five showed a 50-80% reduction in seizures between 2 and almost 5 years following treatment. PMID- 15129841 TI - The effect of antiepileptic drugs on visual performance. AB - Visual disturbances are a common side-effect of many antiepileptic drugs. Non specific retino- and neurotoxic visual abnormalities, that are often reported with over-dosage and prolonged AED use, include diplopia, blurred vision and nystagmus. Some anticonvulsants are associated with specific visual problems that may be related to the mechanistic properties of the drug, and occur even when the drugs are administered within the recommended daily dose. Vigabatrin, a GABA transaminase inhibitor, has been associated with bilateral concentric visual field loss, electrophysiological changes, central visual function deficits including reduced contrast sensitivity and abnormal colour perception, and morphological alterations of the fundus and retina. Topiramate, a drug that enhances GABAergic transmission, has been associated with cases of acute closed angle glaucoma, while tiagabine, a GABA uptake inhibitor, has been investigated for a potential GABAergic effect on the visual field. Only mild neurotoxic effects have been identified for patients treated with gabapentin, a drug designed as a cyclic analogue of GABA but exhibiting an unknown mechanism while carbamazepine, an inhibitor of voltage-dependent sodium channels, has been linked with abnormal colour perception and reduced contrast sensitivity. The following review outlines the visual disturbances associated with some of the most commonly prescribed anticonvulsants. For each drug, the ocular site of potential damage and the likely mechanism responsible for the adverse visual effects is described. PMID- 15129842 TI - Depression and anxiety before and after temporal lobe epilepsy surgery. AB - PURPOSE: To examine the course of depression and anxiety in patients with temporal lobe epilepsy (TLE) treated with epilepsy surgery (and anticonvulsant drugs) or medical means alone. METHODS: Of 94 TLE patients evaluated for epilepsy surgery, 76 underwent a resective procedure, 18 continued on medical treatment alone. Depression (Beck Depression Inventory (BDI)) and anxiety scores (Self Rating Anxiety Scale (SRAS)) were examined during presurgical evaluation (T1) and after a mean of 16 months (T2), or 12 months after surgery. Depression and anxiety scores were related to type of intervention, underlying epileptogenic lesion, change of seizure control and anticonvulsant therapy. RESULTS: At T1, depression and anxiety scores were higher in patients with TLE than scores in published normal populations. At T2, depression but not anxiety scores were significantly lower than at T1. Change of depression scores interacted with improvements of seizure control. CONCLUSIONS: Evidence of depression and anxiety is commonly found in patients with TLE. Depression improves not because of epilepsy surgery per se, but because of improved seizure control. This is more commonly achieved by surgery than medical treatment. The results are consistent with the hypothesis that depression in TLE is caused by pathological epileptic activity rather than a fixed structural defect. PMID- 15129843 TI - Dopamine and cognition. PMID- 15129844 TI - Dopamine: a key regulator to adapt action, emotion, motivation and cognition. AB - Dopamine is expressed in restricted brain areas involved in numerous integrative functions contributing to automated behaviours that are highly adaptive. During ontogenesis, dopamine can have a trophic action, which influences cortical specification directly, especially in prefrontal areas. Such a role is attested by the close relationships existing between the development of dopamine cortical innervation and cognitive abilities. Interestingly, such a proposal is reinforced by phylogenetic data. During the last stages of evolution in mammals, the characteristic extension of dopamine cortical innervation is also correlated with the development of cognitive capacities. More generally, the contribution of dopamine will be to increase the processing of cortical information through basal ganglia, either during the course of evolution or development. In this respect, dysmaturation suspected in schizophrenia and attention deficit-hyperactivity disorder in children can emphasize such a defect in basal ganglia processing. Remarkably, these diseases are improved by dopamine antagonists and agonists, respectively. In this line of evidence, experimental studies showed that selective lesions of the dopamine neurones in rats or primates can actually provide motor, limbic and cognitive deficits, in later cases especially when the mesocorticolimbic dopamine pathways are altered. Data resulting from lesion studies also showed significant alteration in attentional processes, thus raising the question of the direct involvement of dopamine in regulating attention. Dopamine can act as a powerful regulator and integrator of different aspects of brain functions. For example, in Parkinson's disease, besides motor impairment, dopamine degeneration is also expressed by alterations of both limbic, executive and cognitive functions, both improved by dopamine receptor agonists and dopa therapy. Dopamine has thus to be considered as a key regulator that contributes to behavioural adaptation and to the anticipatory processes necessary for preparing voluntary action consequent upon intention. In this respect, the alteration of dopamine transmission with age could contribute to cognitive impairment. Therefore, to normalize dopamine transmission pharmacologically could actually improve the cognitive and limbic deficits during normal aging, as it does in psychiatric and neurological disorders. PMID- 15129845 TI - Dysfunction of the human memory systems: role of the dopaminergic transmission. AB - Human memory is currently considered to consist of different subsystems subserved by different anatomofunctional networks. However, the biochemical basis of memory disturbances remains unclear. The present review focuses on data from studies in Parkinson's disease in order to define the role played by the dopaminergic system in episodic memory and procedural learning. The pattern of episodic memory disturbances in Parkinson's disease is very specific: Parkinson's disease patients display a preserved ability to encode, store and consolidate new information but have difficulties in retrieving it. This deficit is particularly observed when patients have to self-initiate remembering strategies, as they are unimpaired when retrieval cues are explicitly presented. This probably reflects executive dysfunction, and is related to the disruption of the associative striato-prefrontal circuit. It is observed with verbal as well as visuospatial material. On procedural learning tasks, Parkinson's disease patients display a lower progressive performance enhancement over trials than healthy controls. However, this impairment mainly concerns the initial learning phase and appears to be highly dependent on the motor demands of the task. The role played by dopamine depletion in these deficits remains a subject of debate as some studies suggest the involvement of the dopaminergic system in cognitive function, although other results do not support it. Other neurotransmission systems are probably involved in the pathophysiology of cognitive dysfunction in Parkinson's disease, and currently it is the cholinergic hypothesis that is the most considered. PMID- 15129846 TI - Dopamine and cognitive function. AB - PURPOSE OF REVIEW: Although the cognitive deficits of idiopathic Parkinson's disease are now relatively well known, their neuropsychological and neurobiological basis are still discussed. RECENT FINDINGS: As well as recent findings on cognitive changes and their underlying mechanisms, we will review new approaches concerning the effects of levo-dihydroxyphenylalanine, those of deep brain stimulation and recent developments on personality disorders. SUMMARY: Future research directions concern the relative influence of the striatofrontal loops in cognition and personality, and the respective contribution of subcortical and cortical lesions or dysfunctions, both in dementia and in specific cognitive and personality changes. PMID- 15129847 TI - The anti-dementia drugs: myth, hype or reality? AB - The neurodegenerative diseases are in need of drugs that are capable of treating their many different presentations. Some drugs have recently been developed and approved by the authorities for use in Alzheimer's disease; their beneficial effects are no longer questionable. From the discussion about what to call these drugs (anti-dementia drugs?), it is apparent that pharmacology research has veered from pursuing the myth of the fount of eternal youth back to the reality of the struggle for survival of neurons whenever they are attacked, long before the signs of dementia have developed (anti-apoptotic drugs?). The neurosciences teach that there is a biology of cognition (hence a pharmacology of cognition), and that the brain is the permanent area of the confrontation between neurogenesis and apoptosis. Pharmacology has taken this new understanding on board, and has finally defined its objective as preventing the decline of the neuron as much as of cognitive performance. It remains for clinicians to confirm the authenticity of this worldwide project. PMID- 15129848 TI - Working memory and dopamine: clinical and experimental clues. AB - Working memory has been successively considered as a mnesic or executive process. The cognitive processes involved in working memory and the executive functions are closely linked. Most authors currently agree that executive functions include planning, attentional maintenance, mental flexibility and attentional inhibition. Considering that the role of the central administrator, the main module of the working memory model, is to manage new situations, inhibit old non-pertinent schemes, or carry out attentional control, it is clear that it involves the different executive processes mentioned above. Therefore, even though the working memory model has its origins in the classic concept of short-term memory, it is now situated at the interface between memory and executive functions. The identification of the neuroanatomical support of these processes has been widely explored for many years. The involvement of monoaminergic neurotransmitter systems, and in particular of the dopaminergic system, in these complex cognitive functions has been suggested by numerous studies, both in humans and in non-human primates. PMID- 15129849 TI - The role of dopamine in cognition: evidence from functional imaging studies. AB - Functional imaging provides a sensitive means of studying the dopaminergic system in the brain. Both pre-synaptic and post-synaptic aspects of this system can be explored, and recent technical advances even allow the estimation of the synaptic level of dopamine in the striatum. However, only a few studies have used functional imaging to identify the role of dopamine in cognition. This paper reviews recent evidence provided by studies in healthy individuals and patients with Parkinson's disease or schizophrenia supporting the role of dopamine in normal and pathological cognitive processes. PMID- 15129850 TI - Mild cognitive impairment: a nosological entity? AB - Mild cognitive impairment is not an established diagnosis but a concept for which different criteria have been proposed and modified over time. Mild cognitive impairment refers to the transitional zone between normal ageing and dementia. The mild cognitive impairment stage may be one of the optimum stages at which to intervene with preventive therapies. The heterogeneity of the term has been recognized. Multiple sources of heterogeneity have been described for mild cognitive impairment, including biological factors, clinical symptoms and clinical course. It is still not possible to consider mild cognitive impairment as an explicit predictor of Alzheimer's disease with the current criteria. The heterogeneity within mild cognitive impairment has been noted, and a classification has been proposed: amnestic or single memory mild cognitive impairment, multiple domains mild cognitive impairment, and single non-memory mild cognitive impairment. Future research should be focussed on redefining the criteria of the mild cognitive impairment entity. This could enable the better development of appropriate therapeutic interventions. PMID- 15129851 TI - Defining hypertension: what JNC fails to see. PMID- 15129852 TI - A study on plasma selenium level among pregnant women at Rohtak, Haryana. AB - Selenium is an important trace element for human health. Selenium levels in mothers may have significant effects on fetal and neonatal nutrition. In a cross sectional study undertaken in Rohtak, Haryana, plasma selenium level in mothers was investigated at the time of delivery. The mean plasma selenium level in full term mothers (n=82) was 70.63 +/- 1.62 ppb and in preterm mothers (n = 58), it was 63.28 +/- 2.31 ppb. The difference was statistically significant (p < 0.05). These levels were, however, below the reported normal level of 75-120 ppb in adults. Rura/urban stay, parity, maternal age, socio-economic and literacy status etc. did not influence the selenium level of mothers in either of the groups. PMID- 15129853 TI - A comparative study on diagnostic differentials of T3 and T4 hormones for thyroid disorders. AB - Results of Thyroid function tests of 671 individuals have been studied. The analysis shows that T3 and T4 hormone tests are not efficacious as compared to TSH test (p < 0.001) and in diagnosing Hypothyroid cases T3 and T4 lead to almost same results while in detecting Hyperthyroid cases T3 is superior to T4 (p < 0.01). PMID- 15129854 TI - An epidemiological study of ancylostomiasis in a rural area of Kanpur district Uttar Pradesh, India. AB - In this epidemiological study, stool samples were collected from 256 study subjects selected from seven villages of Kanpur district, Uttar Pradesh, India. The average age of the study population was 21.6 years. The overall prevalence rate of hookworm infestation was found to be 34%. Men had significantly higher rate of infestation at all ages. Prevalence rates increased with ages as well. In addition, adult worms were collected from twelve subjects. All were identified as Ancylostoma duodenale. PMID- 15129855 TI - A study of knowledge about malaria and treatment seeking behaviour in two tribal communities of Manipur. AB - The study was conducted on a sample of 250 tribal people of Manipur state, comprised of 125 from the urban valley population and 125 persons from rural tribals settled on hilly area at a higher altitude. The finding shows that knowledge regarding transmission of malaria, self protection and treatment seeking behaviour is still poor among the tribal communities of Manipur. However the urban tribals had better knowledge regarding diagnosis of malaria and prevention of mosquito breeding than their rural counterparts. PMID- 15129856 TI - Some clinico-epidemiological aspect of bronchiolitis among infants and young children--a hospital based study. AB - A hospital based prospective study was carried out from 1st October 1998 to 30th September 1999 on children with clinical diagnosis of bronchiolitis in OPD and indoor patients of the Department of Paediatric Medicine, Medical College Kolkata. The objective of the study was to find out the proportional case rate and clinico epidemiological features of the disease. The effectiveness of nebulized salbutamol among bronchiolitis children was also looked into. The proportional case rate was found to be 4.65%. Low birth weight (LBW)/premature babies (51.11%) malnutrition (40%), nonimmunization (55.55%) & non breastfeeding practices (48.88%) were significant risk factors for severe bronchiolitis. Response to nebulized salbutamol was remarkably higher (70%) in 6m to 12m age group. PMID- 15129857 TI - Biomedical publications in south Asian countries. AB - By and large, biomedical research is not a priority sector in south Asian countries, land of world's one-fifth population. The total number of studies published during 1990-98 from each of the south Asian countries were elicited based on MEDLINE database. The number of studies were normalised by population and physician size, and gross domestic products. The results showed that the following countries are more productive: India and Sri Lanka, when publications were normalised to population; India, Sri Lanka and Nepal when normalised to GDP; Nepal and Sri Lanka when normalised to physician number. PMID- 15129858 TI - An epidemiological study of the risk factors of occupational diseases in coal handling plant of a thermal power station. AB - Anthropometry and morbidity profile of fifty randomly selected workers of coal handling plant at a thermal power station of West Bengal having at least five years experience were studied. The mean value of their age, duration of present job and Body Mass Index (BMI) were 34.8 years, 7.86 years and 19.24 respectively. As they were exposed to coal dust (Av. concentration 300-350 mg/M3 of air), warmth (Ambient temperature 40 +/- 5 degrees C), humidity (Relative humidity 80 +/- 5%) and noise [Av. 80 dB(A)], respiratory system was most commonly (72%) effected followed by greying of hair (5.6%), Cloth dusters used by some could not render any discernable protective effect. Addiction was also prevent (62%). Use of proper protective equipments (PPE) supplemented by adequate preplacement and periodic medical examination followed by successful rehabilitation were suggested. PMID- 15129859 TI - Morbidity profile of aged population of old age home in Calcutta. PMID- 15129860 TI - Is coronary heart disease a communicable disease? PMID- 15129861 TI - Adolescent health. PMID- 15129862 TI - 'Geriatrics' a challenge for the twenty first century. PMID- 15129863 TI - Energy balance of adolescent girls in rural area of Varanasi. AB - Although several studies have been undertaken on dietary intake of adolescent girls, studies on their energy expenditure and thereby energy balance are few. In order to explore these aspects, this community based cross-sectional study was done on 270 adolescent girls of a rural area of Varanasi, selected by adopting appropriate sampling technique. Nutritional status of study subjects was assessed on the basis of BMI. Their energy intake was computed by 24 hours recall oral questionnaire method. Estimation of energy expenditure was based on physical activity which was noted by 24 hours activity recall questionnaire method. Average energy intake (1609.42 +/- 528.87 kcal/day) of the study subjects was less than their mean energy expenditure (1896.19 kcal/day). Their energy expenditure was significantly influenced by several factors (e.g. age, caste, type of family, etc.) As much as 69.63%, subjects were in negative energy balance. Majority of subjects belonging to lower (79.37%) and middle (75.19%) SES were in the negative energy balance. Negative energy balance has been significantly less in subjects belonging to high SES, having main occupation of the family as business and high per capita income groups with respect to their corresponding subcategories. In all 68.52% study subjects had BMI < 18.5 kg/m2. Majority (91.11%) of adolescent girls had rate of energy expenditure > 1.4. Under nutrition was significantly (p<0.05) more in subjects with rate of energy expenditure > 1.4 (i.e. 70.33%) than in subjects with rate of energy expenditure < or = 1.4. PMID- 15129864 TI - Development of a training programme for school teachers on school health service. AB - A training programme was developed and implemented for 56 primary school teachers of Barasat II block in North 24 Parganas district as per their recommended roles and responsibilities in School health service. The methodology consisted of development of learning objectives, evaluation parameters (questionnaire and check list) and teaching-learning materials. This was followed by pre-training assessment, implementation of training and re-training and evaluation of the outcome 7 days and 3 months after initial training and re-training. One way analysis of variance revealed significant overall improvement of the cognitive, affective and psychomotor domain of learning of school teachers. The need of periodic re-inforcement training with adequate scope of practice under supervision were emphasized. PMID- 15129865 TI - Hepatic steatosis and hepatitis C. PMID- 15129866 TI - Steatosis and chronic hepatitis C virus infection. AB - OBJECTIVE: To asses the presence of steatosis and other histological changes in patients with chronic hepatitis C. METHODS: Liver biopsy samples were reviewed for presence of steatosis, its degree and other histological changes of hepatitis C including necro-inflammatory score, fibrosis grade and these changes were correlated with liver function tests. RESULTS: A total of 109 liver biopsy samples were reviewed. Mean age of the patients was 44.46 +/- 13.93 years and 62 (56.8%) were male. Mean necro-inflammatory score was 2.32 +/- 0.95. Mean fibrosis grade was 1.69 +/- 1.12. No Steatosis was found in 42 (38.5%) samples and mild to severe degree of steatosis was found in 67 (61.5%) samples. CONCLUSION: In this study, nearly 62% of liver biopsy samples had some degree of steatosis. Whether this steatosis increases occurrence of severe form of fibrosis or is an associated phenomena with chronic inflammation needs to be further clarified. PMID- 15129867 TI - Quality of life assessment in Pakistani patients with chronic liver disease. AB - OBJECTIVE: To find out if advanced liver disease causes decrement in health related quality of life in Pakistani patients. METHODS: An observational study was conducted at the Shifa International Hospital Islamabad. Chronic Liver Disease Questionnaire (CLDQ) was administered to patients with liver disease who were either admitted or seen as outpatients. Patients were divided into two groups. Those with no cirrhosis or Child's Class A were placed into group A. Those with advanced liver disease i.e. Child's Class B and C into group B. RESULTS: A total of 56 patients (mean age 50.6 years) were included in this study. Hepatitis C was the cause of chronic liver disease (CLD) in 52 (92.9%) patients. Twenty-one (37.5%) patients were in group A and 35 (62.5%) in group B. Patients with advanced liver disease had significantly lower CLDQ scores as well as decrement in five out of the six categories for assessing quality of life. CONCLUSION: Chronic liver disease causes significant impairment in the Quality of Life Index (QLI) in Pakistani patients. PMID- 15129868 TI - Knowledge and practices of barbers about hepatitis B and C transmission in Rawalpindi and Islamabad. AB - OBJECTIVE: To assess the knowledge and practices of barbers regarding transmission risk of HBV and HCV viruses. METHODS: A cross-sectional survey of barber's shops in Rawalpindi and Islamabad was conducted during September- November1998. Barbers were queried about hepatitis, knowledge regarding hepatitis transmission through razor, vaccination, sterilization, and the form of media they use for information and entertainment. Use of instruments on at least 2 clients were observed in each shop. Proportion and their 95% confidence intervals were computed. RESULTS: Of 96 barbers approached, 12 (13%) knew that hepatitis is a disease of the liver, causing jaundice, it is transmitted through parenteral route and could also be transmitted by razor. During the actual observation of 192 clients, razors were cleaned with antiseptic solution for 22 (11.4%) and reused for 88 (46%) shaves. CONCLUSION: Level of awareness among barbers about hepatitis and risks of transmission is very low, and their practice of razor reuse that may spread hepatitis is very common. Messages about hepatitis need to be incorporated in media campaigns, in addition to regulation of practices. PMID- 15129869 TI - Etiology of chronic liver disease in children. AB - OBJECTIVE: To review the clinical spectrum and etiology of chronic liver disease in children at National Institute of Child Health, Karachi. METHODS: Prospective study in children aged 1 to 14 years with suspected chronic liver disease. Complete blood count, LFT's, prothrombin time, serum albumin and ultrasound of abdomen were done in all patients. Liver biopsy was done in majority of the cases. Viral markers included HBsAg, anti HCV and ANA to determine etiology. Those who were negative for hepatitis B, C and autoimmune disease, were subjected to slit lamp examination of eyes and 24 hours urinary copper estimation for Wilson's disease. Alpha-1-antitrypsin levels were done in selected patients. RESULTS: A total of 55 cases were studied. The common presenting features were edema, ascites (44 patients), jaundice (27), variceal bleeding (23) and fever (22). On examination anemia was present in 52 patients, edema in 46, jaundice in 37, splenomegaly in 42 and hepatomegaly in 35 patients. Fortynine patients had hypoalbuminemia (< 2.5 gm%), 45 raised ALT (> 80 IU/L) and 49 prolonged PT (> 4 sec of control). Ultrasonography showed a dilated portal vein in 34 patients and esophageal varices were seen in 46 patients on endoscopy. Thirteen (24%) had chronic hepatitis B, 9(16%) autoimmune disease, 9 (16%) Wilson's disease and all were anti HCV negative. Etiology remained uncertain in 24 (44%) cases. CONCLUSION: Hepatitis B was the leading cause of chronic liver disease in children followed by Wilson's disease and autoimmune liver disease. None of the patients had hepatitis C in this study. PMID- 15129870 TI - The evaluation of hepatocellular carcinoma with biphasic contrast enhanced helical CT scan. AB - OBJECTIVE: To evaluate the role of biphasic contrast-enhanced helical CT including Hepatic Arterial Phase (HAP) imaging with Portal Venous Phase (PVP) imaging, in the detection and characterization of hepatocellular carcinomas. METHODS: The study included 40 patients (M = 26, F = 14) with histopathologically proven HCC. Age range was between 30-85 years (mean = 55) by following consecutive patients with cirrhosis in whom Hepatocellular carcinoma was diagnosed or suspected either by raised serum a (alpha)-fetoprotein level or Ultrasound. RESULTS: Biphasic contrast-enhanced examination revealed a total of 85 lesions in these 40 patients, out of which 13 were unifocal, 12 showed a dominant mass with satellite lesions, 2 showed cluster of contiguous nodules and 13 were multifocal HCCs. Mean diameter was 3.1 cm, ranging from 0.8 to 14 cm. On HAP imaging 85% were detected. (hyperattenuating = 69, hypoattenuating = 3) while on PVP imaging detectibility was only 48% (hyperattenuating = 2, hypoattenuating = 39). Hence detectibility was significantly (p = 0.008) superior in HAP as compared to PVP imaging. In 7 patients (17%) tumor was visible only on HAP images. Venous invasion was present in 12 patients (30%) while arterio-portal shunting was seen in 5 patients (13%). CONCLUSION: Biphasic contrast enhanced helical CT is a useful method in detection and characterization of HCC. PMID- 15129871 TI - The role of magnetic resonance cholangiopancreatography (MRCP) in obstructive jaundice. AB - OBJECTIVE: To evaluate the diagnostic value of MRCP in studying the sites and cause of obstructive jaundice in comparison with other imaging modalities at the Department of Radiology, Aga Khan University Hospital, from January 1999 to May 2001. METHODS: Forty nine consecutive patients included 19 men and 30 women, suspected of obstructive jaundice. Patients underwent ultrasound (n = 49), CT (n = 11), ERCP (n = 25) and biliary surgery (n = 17). Final diagnosis was established by surgical exploration, endoscopic sphincterectomy, cytology and clinical follow up. RESULTS: Of the 49 patients 17 had choledocholithiasis. Twenty five patients had malignant strictures, out of which 11 had non-specific malignant strictures, 7 had pancreatic carcinoma, 3 had Klatskin tumors, 3 had periampullary carcinoma and 1 had gallbladder carcinoma. Six patients had benign strictures and 1 patient had choledochal cyst. Overall, MRCP was sensitive (88%) and specific (96.8%) in detecting choledocholithiasis. MRCP sensitivity and specificity in detecting benign main bile duct stricture was equal to 83.3% and 97.6% respectively, and 92% and 100% for malignant stricture. CONCLUSION: Our prospective study confirms that MRCP, a noninvasive and well tolerated imaging technique is of value in the diagnosis of obstructive jaundice. PMID- 15129872 TI - Tropical sprue: revisited. AB - OBJECTIVE: To review the experience of patients presenting with clinical manifestations of tropical sprue and assess their diagnosis and management, response to treatment and follow up. METHODS: This single center retrospective descriptive study was done at Shifa International Hospital, Islamabad from January 1994 to January 2003. All patients who presented with diarrhea, anorexia, weight loss and anemia and who proved to have partial villous atrophy on intestinal biopsy and had responded to treatment with antibiotic and folic acid,were included in this review. RESULTS: A total of 42 patients were encountered during these years. There were 31 (74.0%) males and 11 (26%) females. The age ranged from 17-66 years. All patients presented with diarrhea, weight loss, anorexia and had megaloblastic anemia. In all patients, a distal duodenal biopsy showed partial villous atrophy. All were treated with tetracycline 1 g per day and folic acid 5 mg per day and responded to treatment within 4 weeks. Total treatment lasted 3 months and resulted in complete resolution of symptoms and gain of weight. The follow up lasted for a mean of 5 years and no relapses were noted. CONCLUSION: Tropical sprue presents with diarrhea, anorexia, weight loss, and megaloblastic anemia. The partial villous atrophy has been a constant finding. The response to treatment to tetracycline and folic acid has been uniformly successful. PMID- 15129873 TI - Seroprevalence of hepatitis C virus infection and its risk factors in pregnant women. PMID- 15129874 TI - Esophageal cancer--a review. PMID- 15129875 TI - Transcatheter chemo-embolization for hepatocellular carcinoma and certain hepatic metastasis. PMID- 15129876 TI - PSG consensus statement on management of hepatitis C virus infection--2003. PMID- 15129877 TI - PGS consensus statement on management of hepatitis B virus infection--2003. PMID- 15129878 TI - Esophageal foreign body in neonates. PMID- 15129879 TI - Paraduodenal hernia--a case report. PMID- 15129880 TI - Obstructive jaundice: an unusual presentation of a superior mesenteric artery aneurysm. PMID- 15129881 TI - Motor neuronopathy associated with adenocarcinoma of esophagus. PMID- 15129882 TI - Statistical considerations for use of composite health-related quality-of-life scores in randomized trials. AB - BACKGROUND: Quality of life instruments are frequently used as outcomes in randomized trials. Instruments that consist of several subscales present researchers with a choice of whether to combine some or all scales into a single composite score. There may be several clinically and scientifically reasonable alternative combinations of subscales for the primary outcome measure. MAJOR FINDINGS: The statistical efficiency of different combinations of subscales depends on the relative effect size of the intervention on each subscale and the correlation between the subscales. Simple equations can be derived for determining the relative statistical efficiency of each clinically reasonable combination of subscales. Hypothetical scenarios show that the number of patients needed in a clinical trial can be twice as great for some combinations of subscales as for others. CONCLUSIONS: There are often compelling clinical or scientific reasons to use a particular subscale or composite in a randomized trial. In the case where a number of different alternatives would be reasonable, statistical efficiency can help guide the choice of endpoint. PMID- 15129883 TI - Interpreting differences in quality of life: the FACT-H&N in laryngeal cancer patients. AB - Quality of life (QOL) scores can be difficult to interpret, because small statistically significant differences can be clinically unimportant. Our goal was to estimate the magnitude of difference in QOL that is noticeable to patients. METHODS: Laryngeal cancer patients (n = 98, male = 83%, mean age = 65) completed a QOL questionnaire, FACT-H&N. Paired participants rated their own QOL as compared to each other. We estimated the smallest difference in QOL score that was associated with a noticeable difference in patients' subjective ratings. RESULTS: Differences in FACT-H&N score were somewhat correlated with patients' ratings of their well-being relative to other patients (r = 0.195, p < 0.0001). The FACT-H&N score had to differ by 6.22 for patients to rate themselves as 'a little bit better' relative to other patients (95% CI: 1.42-11.02), and by 12.40 for patients to rate themselves as 'a little bit worse' relative to others (95% CI: 5.09-19.71). Results were consistent regardless of patient age, gender or laryngeal subsite but were imperfect predictors of individual judgements. CONCLUSION: The minimal important difference for the FACT-H&N score is about 6-12 units, but laryngeal cancer survivors may be more sensitive to gains than losses. PMID- 15129884 TI - A cardiac-specific health-related quality of life module for young adults with congenital heart disease: development and validation. AB - This study represents the development and validation of a cardiac-specific module of the generic health-related quality of life (HRQoL) instrument, the TAAQOL (TNO/AZL Adult Quality Of Life), for young adults with congenital heart disease (CHD). Items were selected based on literature, an explorative previous study in CHD patients, interviews with patients, and the advice of experts. The newly developed Congenital Heart Disease-TNO/AZL Adult Quality of Life (CHD-TAAQOL) was tested in 156 patients with mild or complex CHD and consisted of three hypothesised subject scales: 'Symptoms' (9 items), 'Impact Cardiac Surveillance' (7 items), and 'Worries' (10 items). Cronbach's alpha for the three scales were 0.77, 0.78, and 0.82, respectively. Scale structure was confirmed by Principal Component Analysis, corrected item-scale and interscale correlations. Overall, 55% of reported health status problems were associated with negative emotions, which is an argument for assessing HRQoL as a concept distinct from health status. Convergent validity with validated generic instruments (TAAQOL and Short Form-36, SF-36) showed satisfactory coefficients. Discriminant validity was proven by significantly higher scores for mild CHD patients compared with those with complex CHD. In conclusion, the CHD-TAAQOL module together with the generic TAAQOL can be used to assess group differences for cardiac-specific HRQoL in young adults with CHD. Testing psychometric properties of the CHD-TAAQOL shows satisfactory results. However, to detect changes in HRQoL over time, further research is needed. PMID- 15129886 TI - Quality of life as conveyed by pediatric patients with cancer. AB - Quality-of-life instruments have provided important advances in measuring the quality of life of pediatric patients receiving treatment for cancer. However, the bases of these instruments have not included first-hand reports from the patients; thus, these instruments may be conceptually incomplete. We directly solicited from pediatric patients their perspectives regarding their quality of life during treatment for cancer. We conducted two pilot studies: 23 patients (aged 8-15 years) participated in the first, a cross-sectional study; and 13 patients (aged 10-18 years) participated in the second, a 2-year longitudinal study. Data were analyzed by using a semantic-content method, and the following six domains were recognized in data from both of the studies: symptoms, usual activities, social/family interactions, health status, mood, and the meaning of being ill. These domains were compared with those of seven established pediatric oncology quality-of-life instruments, none of which included all six of these domains; the domain most frequently missing was the meaning of being ill domain. Here we present a new definition of the quality of life of pediatric oncology patients that is based on six domains; this definition may ensure the completeness and sensitivity of these important instruments. PMID- 15129885 TI - Construction and validation of a quality of life instrument for young adults. AB - Assessment of quality of life (QOL) in young adults has become an increasingly important field of research as information on the QOL of children and adults accumulates. The purpose of this study is to report on the development of the Young Adult Quality Of Life (YAQOL) instrument, a measure of QOL for young adults aged 18-25. The YAQOL, which is comprised of 14 multi-item scales that assess physical health and aspects of psychological well-being, social relationships, role function, and environmental context, was administered to a general population sample of 751 young adults. Scale structure was confirmed by item internal consistency, item discriminant validity, and inter-scale correlations. Reliability coefficients ranged from 0.88 to 0.63 across scales (mean = 0.73). Eight YAQOL scales discriminated clearly between young adults with and without chronic physical conditions, 12 scales discriminated between young adults with and without personality disorder, and negative associations were demonstrated between YAQOL scale scores and psychiatric disorder symptoms. Demographic differences in the YAQOL scales were consistent with theoretical expectations and previous empirical work. Overall, findings support the reliability and utility of the YAQOL as a measure of QOL in young adults in the general population. Additional psychometric properties will continue to be evaluated as more data become available. PMID- 15129887 TI - Health status versus utilities of patients with end-stage liver disease. AB - BACKGROUND: Health-related quality of life (HRQL) in patients with end-stage liver disease (ESLD) can be evaluated using either health-status questionnaires or utility assessment techniques. The two approaches have never been compared in terms of the values they assign to health prior to liver transplantation. STUDY DESIGN: We assessed health status of patients with ESLD using validated disease specific instruments covering multiple domains (measures of disease, psychological status, personal function, social/role function, and general health perception). We also elicited utilities using formal approaches (standard gamble [SG] and time tradeoff [TTO]) and a simpler alternative (visual analog scale [VAS]). PATIENTS: Outpatients and inpatients at a single center prior to liver transplantation (n = 78). PRINCIPAL FINDINGS: Health status was generally poor (median physical symptoms score on a 0-1 [worst to best] scale, 0.33; psychological symptoms, 0; happiness, 0.50; personal function, 0; social/role function, 0.40; and general health perception, 0.40). The median VAS score was 0.50. The median TTO was 0.79, indicating that half of the patients in our sample chose healthier life in return for a 21% shorter life expectancy. The median SG score was 0.50, indicating that half of the patients were willing to take up to a 50% risk of death in exchange for perfect health. CONCLUSIONS: Both health status measures and utility assessments indicate that HRQL is compromised in patients awaiting liver transplantation. Despite the overall consistency between the two approaches, however, health status measures do not serve as reasonable proxies for utilities. For formal economic evaluations such as cost effectiveness analyses, only direct measures of utility can be used to quantify health states. PMID- 15129888 TI - Perceived quality of life in schizophrenia: relationships to sleep quality. AB - We examined the relationship between perceived quality of life (QOL) and subjective quality of sleep among schizophrenia patients, and its relation to symptom severity, side effects and emotional distress. One hundred and forty five schizophrenia patients were comprehensively evaluated with standardized measures of symptom severity, adverse effects, emotional distress, QOL, and sleep quality (the Pittsburgh Sleep Quality Index, PSQI). Partial correlation and multiple regression analyses were performed. Poor sleepers reported lower mean scores on all QOL domains, they were more depressed and distressed, and had more adverse effects to medications with concomitant distress than good sleepers. The negative relationship between complaints of poor sleep quality and QOL measures remained significant when the confounding effect of depression, side effects, and distress was partialled from the correlation matrix. Daytime dysfunction (a component of the PSQI) accounted for 12.6% of the variance in QOL index scores. Thus, poor QOL reported by schizophrenia patients is substantially associated with poor sleep quality. This association appears both independently and synergistically with depression, distress and side effects of medications. PMID- 15129890 TI - A comparison of the discriminatory power of the Inflammatory Bowel Disease Questionnaire and the SF-36 in people with ulcerative colitis. AB - The aim of this study was to compare the discriminatory power of a generic and a condition-specific quality of life measure (the Medical Outcomes Study Short-Form 36 (SF-36) and the Inflammatory Bowel Disease Questionnaire (IBDQ)) with respect to sub-groups defined by disease-related and other factors, in a sample of patients with ulcerative colitis. Disease activity was generally more highly correlated with IBDQ scores than with SF-36 scores. The only significant differences with respect to disease extent were in the SF-36 energy/vitality and social function domains. Age was negatively and weakly to moderately correlated with the physical domains of the SF-36 but positively though weakly correlated with scores on the IBDQ emotional domain and those domains of the SF-36 related to mental well-being. Co-existing chronic illness, even on controlling for age, was associated with significantly poorer scores on the generic measure, but had little influence on IBDQ scores. In conclusion, generic and disease-specific measures of quality of life appear to be complementary rather than interchangeable. We recommend the use of both types of measure in parallel. PMID- 15129889 TI - Health-related quality of life in cardiac patients with dyslipidemia and hypertension. AB - BACKGROUND: Hypertension is associated with lower health-related quality of life (HRQOL). Similar association may be found for dyslipidemia. However, controversies exist regarding the HRQOL with dyslipidemia. We evaluated the HRQOL of cardiac patients with and without dyslipidemia and hypertension. METHODS: In a cross-sectional study, 284 cardiac patients rated their HRQOL using SF-36 Health Survey (SF-36), and three preference-based measures (Rating Scale, Time Trade-off and Standard Gamble). RESULTS: Compared to those without dyslipidemia, those with dyslipidemia reported better HRQOL on all preference-based measures and most SF 36 scales particularly on the physical health scales. Adjusted mean differences and 95% confidence interval (95% CI) were equal to 4.5 (0.5, 8.5), 10.8 (2.8, 18.8), and 2.2 (0.2, 4.2) on the Physical Functioning, the Role-Physical and the Physical Component Summary scales, respectively. Exactly the opposite trends were observed among patients with hypertension. The adjusted mean differences (95% CI) were equal to -2.7 (-6.7, 1.4), -10.9 (-19.1, -2.8), and -2.9 (-4.9, -0.9) on the Physical Functioning, the Role-Physical and the Physical Component Summary scales, respectively. CONCLUSION: Cardiac patients with hypertension reported lower physical health than those without hypertension while cardiac patients with dyslipidemia reported better physical health than those without dyslipidemia. The reason for these different trends is not known. Possible explanations are discussed. PMID- 15129891 TI - A national study of the importance of oral health to life quality to inform scales of oral health related quality of life. AB - OBJECTIVES: To assess the content validity of traditional oral health status scales as measures of oral health related quality of life (OHQOL), based on a general public's perception of the most important ways in which oral health affects quality of life (QoL). METHODS: A nationwide United Kingdom study involving a random probability sample of 1778 adults. Data were collected by face to-face interviews in participants homes. RESULTS: Most 75% (1332) perceived oral health as being important to QoL. Among them, 53% (699/1332) identified oral health's importance to QoL as being in a positive manner. Existing scales predominantly fail to include this dimension. The general public ranked oral health's importance to QoL through a range of physical, social and psychological domains. Most frequently though affecting eating or comfort; domains considered by all instruments. Other domains/ways are presented. It is apparent that even when some of the multidomain scales are employed, they frequently omit items that the public perceives as being most important to QoL. CONCLUSION: The study raises concerns about the appropriateness of utilising many of the existing oral health status scales as measures of OHQOL because the concepts do not appear to be interchangeable. PMID- 15129892 TI - Development of a preliminary diabetes dietary satisfaction and outcomes measure for patients with type 2 diabetes. AB - Adopting dietary lifestyle changes for diabetes management is often difficult for patients; yet the health-related quality of life (HRQOL) outcomes of dietary management for the patient are not extensively developed in the HRQOL assessments now widely used in diabetes research. This study developed a preliminary instrument, the diabetes dietary satisfaction and outcomes measure, to assess outcomes of individuals' experiences in following a meal plan for the treatment of type 2 diabetes. A theoretical framework and preliminary focus group data guided the design of a 47-item questionnaire, administered to 239 patients with type 2 diabetes. Medical file data was obtained on 180 of these patients. Fifty four percent of respondents were women, with mean age of 64 +/- 12 years and diabetes duration of 10 +/- 8 years. Scores for the satisfaction and other outcome measures discriminated between patient groups by age, gender, medication use, depression diagnosis, meal plan status, and employment status. Significant correlations also occurred with diet adherence, number of co-morbidities, and glycemic control as measured by glycolated hemoglobin (HbA1c). Future research with additional patient samples is needed to refine the measure for use in diabetes education programs. PMID- 15129893 TI - Gastroparesis Cardinal Symptom Index (GCSI): development and validation of a patient reported assessment of severity of gastroparesis symptoms. AB - BACKGROUND: Patient-rated symptom assessments are needed for evaluating the effectiveness of medical treatments and for monitoring outcomes in gastroparesis. OBJECTIVE: This paper summarizes the development and psychometric evaluation of a new instrument, the Gastroparesis Cardinal Symptom Index (GCSI), for assessing severity of symptoms associated with gastroparesis. METHODS: The GCSI was based on reviews of the medical literature, patient focus groups, and interviews with clinicians. A sample of 169 patients with a documented diagnosis of gastroparesis participated in the psychometric evaluation study. Patients completed the GCSI, the SF-36 Health Survey, and disability days questions at baseline and after 8 weeks. A randomly selected sub-sample of 30 subjects returned at 2 weeks to assess test retest reliability. Clinicians rated severity of symptoms, and both clinicians and patients rated change in gastroparesis-related symptoms over the 8 week study. RESULTS: The GCSI is based on three subscales: post-prandial fullness/early satiety (4 items); nausea/vomiting (3 items), and bloating (2 items). Internal consistency reliability was 0.84 for the GCSI total score and ranged from 0.83 to 0.85 for the subscale scores. Two week test retest reliability was 0.76 for the total score and ranged from 0.68 to 0.81 for subscale scores. Construct validity was supported, given that we observed significant relationships between clinician assessed symptom severity and GCSI total score, significant differences between gastroparesis and dyspepsia patients (n = 760) on GCSI total (p < 0.0001) and subscale scores (p < 0.03 to p < 0.0001), moderate and significant relationships between GCSI total and SF-36 scores, and significant associations between GCSI total score and reports of restricted activity and bed disability days. Patients with greater symptom severity, as rated by clinicians, reported more symptom severity on GCSI total score. GSCI total scores were responsive to changes in overall gastroparesis symptoms as assessed by clinicians (p < 0.0001) and patients (p = 0.0004). CONCLUSION: The findings of this study indicate that the GCSI is a reliable and valid instrument for measuring symptom severity in patients with gastroparesis. PMID- 15129894 TI - The impact of subject/respondent characteristics on a proxy-rated quality of life instrument for the Japanese elderly with dementia. AB - The development of a quality of life (QL) instrument for evaluating quality of services requires extensive validation. This study examines the impact of subject/respondent characteristics on a newly developed quality of life instrument for the Japanese elderly with dementia (QLDJ) as a part of the validity examination. In the QLDJ, QL is defined as a three-dimensional construct: 'interacting with surroundings', expressing self', and 'experiencing minimum negative behaviors'. Thus the QLDJ is a multi-dimensional, 24-item instrument, and it is answered by a proxy, the formal caregiver (respondent) because the elderly with severe dementia cannot be directly interviewed. Altogether 623 elderly persons experiencing dementia from 37 institutions (e.g., nursing homes, day care centers) in Japan were assessed using the QLDJ by their formal caregivers. In addition to the levels of dementia and independence in activities of daily living, factors such as gender of the elderly person and respondent, elderly person's age, participation in recreational activities, qualification of the respondent were significantly associated with 'interacting with surroundings' and 'expressing self'. The levels of dementia and independence as well as qualification of the respondent were associated with 'experiencing minimum negative behaviors'. Possible reasons for these associations and how to deal with them are discussed. Careful control for those factors is needed when the QLDJ is used in future research. PMID- 15129895 TI - Cross-cultural adaptation and validation of the Korean fibromyalgia impact questionnaire in women patients with fibromyalgia for clinical research. AB - Our aim was to translate and cross-culturally adapt the fibromyalgia impact questionnaire into Korean (KFIQ), and then evaluate its reliability and validity. The FIQ was translated into Korean by three translators and then independently translated back into English by three different translators. A total of 62 women patients with fibromyalgia (FM) were studied for the psychometric properties of the KFIQ. The mean age of the patients was 47.1 (25-73) years, and all were female. The mean KFIQ score was 48.3 (17-91), and the mean Korean health assessment questionnaire (KHAQ) score was 0.4 (0-1.7). The test-retest reliability of the KFIQ yielded an intraclass correlation coefficient of 0.85 (0.53-0.96). For the construct validity, the Spearman rank correlations of KFIQ with patient global assessments using visual analog scale (pain, 0.58; morning stiffness, 0.45; fatigue, 0.48; depression, 0.43; anxiety, 0.56; global well being, 0.46; disease severity, 0.49; impact on life, 0.51), KHAQ (0.44), and tender points (0.60) were high and statistically significant. The KFIQ might be a reliable and valid instrument for measuring health status and physical functioning in Korean women patients with FM, but needs further study. PMID- 15129896 TI - Validation of the Korean version of the EORTC QLQ-C30. AB - This study evaluated the Korean version of the EORTC QLQ-C30 (version 3.0) in terms of psychometric properties and its validation. One hundred and seventy patients completed three questionnaires EORTC QLQ-C30, the Beck depression inventory (BDI), and a brief pain inventory (BPI). Multitrait scaling analyses demonstrated that all scales met multidimensional conceptualization criteria, in terms of convergence and discrimination validity. Cronbach's alpha coefficients for eight multiple-item scales were greater than 0.70, with the exception of cognitive functioning. All interscale correlations were statistically significant in the expected direction (p < 0.01). Multivariate analyses showed that physical and emotional functioning were significant explanatory variables for the global quality-of-life (QOL) scale (regression coefficients: 0.36, p < 0.001; and 0.37, p < 0.001; respectively). All scales were significantly associated with pain severity and interference of the BPI, and with the cognitive-affective and somatic scales of the BDI. The emotional-functioning scale was substantially correlated with the cognitive-affective scale and somatic scale of the BDI. These results demonstrate that the Korean version of the EORTC QLQ-C30 is a valid instrument for evaluating Korean-speaking patients with cancer, and can be used to distinguish clearly between subgroups of patients of differing performance status. PMID- 15129897 TI - Fifth Annual David A. Winston Lecture. PMID- 15129898 TI - Crossing the quality chasm: implications for health services administration education. AB - Programs in Health Services Administration (HSA) should respond to the mandate to improve patient care as put forth by the Institute of Medicine (IOM) and other reports on the proliferation and consequences of medical errors. This article will identify a framework to base curriculum change, competency areas, and educational methods to impart quality improvement knowledge and skills. The first six competency areas reflect the six redesign imperatives from the IOM report, Crossing the Quality Chasm: A New Health System for the 21st Century (2001): redesign of the care process; use of information technologies; knowledge and skills management; development of effective teams; coordination of care; and use of performance and outcomes measurement. Based on a literature review, five additional areas were identified: strategic quality planning; programs for patient safety and risk management; change management; roles of stakeholders, payers and regulators; and development of a learning environment and blame-free culture. Examples of curriculum content are provided from HSA programs at Georgetown University, University of Washington, and University of California at Berkeley. PMID- 15129899 TI - Teaching economics in the graduate health administration curriculum: a mini-case simulation approach. AB - Training in the discipline of economics has long been recognized as an important component of the curriculum in graduate health administration (GHA) education. Yet economics is a subject that is often considered hard to teach and difficult for students to learn. What is missing is a body of literature that identifies methods that are most appropriate for teaching economics to graduate students aspiring to be health care managers. Moreover, given the keen interest in developing competency-based curricula today, what is also needed is empirical evidence, which systematically links the key skills, knowledge, and abilities economics courses try to develop with the teaching approaches best suited to do so. This paper helps fill this gap in the literature by describing and evaluating the author's "Top of the News" (TOTN) mini-case simulation series piloted in her Health Care Economics course in the Master of Health Administration (MHA) curriculum at Saint Louis University during the spring semester of 2002. This approach combines elements of the case study and simulation methods used by others, but adds several innovations. Although the TOTN method was designed specifically for a course in health care economics, it is one that can be readily adapted for courses across the GHA curriculum. The article concludes that although the TOTN exercises proved to be fairly successful, GHA faculty and practitioners concerned about preparing today's students to meet tomorrow's health care leadership challenges need to not only identify effective teaching methods, but also have a lot more to discover about the learning styles of the students they teach. PMID- 15129900 TI - Case analysis online: a strategic management case model for the health industry. AB - Despite the plethora of methods and tools available to support strategic management, the challenge for health executives in the next century will relate to their ability to access and interpret data from multiple and intricate communication networks. Integrated digital networks and satellite systems will expand the scope and ease of sharing information between business divisions, and networked systems will facilitate the use of virtual case discussions across universities. While the internet is frequently used to support clinical decisions in the healthcare industry, few executives rely upon the internetfor strategic analysis. Although electronic technologies can easily synthesize data from multiple information channels, research as well as technical issues may deter their application in strategic analysis. As digital models transform access to information, online models may become increasingly relevant in designing strategic solutions. While there are various pedagogical models available to support the strategic management process, this framework was designed to enhance strategic analysis through the application of technology and electronic research. A strategic analysis framework, which incorporated internet research and case analysis in a strategic managementcourse, is described alongwith design and application issues that emerged during the case analysis process. PMID- 15129901 TI - U.S. Army-Baylor University Health Care Administration Program: evidenced-based outcomes in the military health system. AB - The purpose of this research is to assess the impact of an educational program on the Military Health System on some of the evidence-based educational outcomes for the Individual (student) and the Society (all Army Medical Treatment Facilities). The U.S. Army-Baylor University HCA program provides a unique opportunity to assess the impact of an educational program on the Military Health System (MHS). Since the majority of the graduate students are military officers who serve in military medical treatment facilities (MTFs), tracking their career progression allows assessing the value added of the U.S. Army-Baylor University HCA experience from 1951 to 2001 (n = 2234). The context of Society outcomes includes all the Army MTFs where U.S. Army-Baylor University HCA graduates execute their leadership skills. During the time from 1994 to 2001, all of the Army MTFs in the MHS (n = 38) were examined by the Joint Commission on Accreditation of Healthcare Organizations (JCAHO). In a similar but shorter time frame (1997-2001), DoD patient satisfaction assessments were conducted. The Individual outcomes (career advancement, increase in status, higher professional association membership) demonstrate that the selection criteria used for program admission appear to be successful. The Society outcomes showed higher JCAHO scores and satisfied consumers in Army facilities with Baylor graduates as the Deputy Commander for Administration (DCA). Continued internal program assessments (curriculum reviews) and external reviews (Accrediting Commission on Education for Health Services Administration accreditations of 5 years in 1987, 8 years in 1993 and 7 years in 2001, and 7 ACHE student chapter awards) attest to the strengths of the U.S. Army Baylor University HCA program. Educating the MHS shareholders (patients, beneficiaries, professional and support staff, senior leaders) and leveraging technology to. share best practices for all administrators (including non-Baylor graduates) will help improve the quality patient care of the Military Health System and other healthcare systems. PMID- 15129902 TI - Trend analysis of minority student enrollment in graduate health management education programs: implications for the field. AB - The need for broad representation of minorities in healthcare management has been an on-going concern for healthcare administrators and professional organizations such as the National Association of Health Services Executives (NAHSE), and the American College of Healthcare Executives (ACHE). Despite the best intentions of healthcare organizations, many have failed to attain a desired racial and ethnic balance in the composition of their senior-level management ranks. This problem will persist unless there is a sustained and collaborative effort between the healthcare professional associations and organizations and the educational programs, especially programs affiliated with the Association of University Programs in Health Administration (AUPHA). Using enrollment data published in the AUPHA directory, this paper examines the enrollment trends of minority students in graduate programs in health management education. The results of this investigation reveal an upward trend in minority student enrollment and that graduate programs with high minority enrollment are located in geographical areas with high minority populations. Suggestions to further increase and sustain minority student enrollment in graduate health management educational programs are advanced. PMID- 15129903 TI - Gallbladder carcinoma in the "Hospital de Clinicas" of Uruguay: 1998-2002. A clinicopathologic study of five cases in 802 cholecystectomies. AB - Five cases of primary carcinoma of the gallbladder are presented. The cases were identified after a study of 802 cholecystectomies in a period of 5 years. The patients are three women and two men between the ages of 43 and 60 years (mean, 55.8 years). In three cases the clinical diagnosis was that of carcinoma, while in two other patients the clinical diagnosis was that of acute cholecystitis. Grossly, all cases were characterized by a gray-white diffuse or focal plaque like thickening of the gallbladder wall, with loss of the normal velvety mucosal surface and fibrosis of the organ. Histologically, four cases belong to moderately to poorly differentiated adenocarcinoma and were characterized by infiltrative, irregularly shaped and sized glands, islands, nests, and cords. The cells showed pleomorphic nuclei with clumped chromatin and frequent single nucleoli. One case was a mucinous adenocarcinoma characterized by large pools of mucoid material with neoplastic glands and cells "floating" within. Pathologic staging was pT3 in three cases; pT2 in one case; and pT2N1 in one other case. The present study highlights the importance of careful gross and histopathologic evaluation of gallbladders otherwise removed with the history of chronic or acute cholecystitis. In addition, it highlights the incidence of gallbladder carcinoma in a particular institution. PMID- 15129904 TI - Co-downregulation of PTEN, KAI-1, and nm23-H1 tumor/metastasis suppressor proteins in non-small cell lung cancer. AB - The multistep process of carcinogenesis implies the accumulation of multiple molecular defects. Alteration of tumor suppressor and metastasis suppressor genes are the important steps. Increasing experimental evidence indicates that decreased expression of tumor-metastasis/suppressor genes and gene products are involved in the progression of a variety of human malignancies. In the present study, we have extended this analysis to non-small cell lung carcinomas (NSCLC). The expression and prognostic significance of the tumor suppressor gene PTEN and metastasis suppressor genes nm23-H1 and KAI-1 was evaluated in NSCLCs. Immunohistochemical staining was performed on formalin-fixed, paraffin-embedded tissues from 53 bronchogenic adenocarcinomas and 51 squamous cell carcinomas using monoclonal antibodies against PTEN, nm23H-1, and KAI-1 proteins. Immunohistochemical results were correlated with tumor stage, grade, lymph nodes positivity, metastasis, and patient survival. Significant co-expression of PTEN, nm23-H1 and KAI-1 was observed in NSCLC (P<.001 to .002). The immunohistochemical expression of these proteins was significantly higher in stages 1 and 2 compared with stages 3 and 4 (P=.04 for PTEN and KAI-1, P=.039 for nm23-H1). When all stages were considered together, loss of immunoreactivity for PTEN, nm23-H1 and KAI-1 was found in advanced NCSCLs (P=.015 for PTEN, P=.001 for KAI-1, P=.004 for nm23-H1), which is suggestive of co-downregulation of these proteins in the process of tumor progression. On multivariate analysis, negative staining for PTEN (P=.014), KAI-1 (P=.034), and nm23-H1 (a trend toward association for nm23 H1 reached near significance P=.08) correlated with disease-related death. Positive lymph node status was associated with negative immunostaining for PTEN (P=.007) but no correlation was observed for nm23-H1 and KAI-1. Loss of expression was linked to distant metastasis (P=.006 for PTEN, P=.002 for nm23H1, P=.001 for KAI-1). On multivariate analysis, co-downregulation of PTEN (P=.009), KAI-1 (P=.02), and nm23-H1 (P=.011) independently predicted shortened survival in NSCLC. Although NSCLC exhibits strong co-expression of PTEN, nm23-H1 and KAI-1, there is a loss of these proteins in high-stage tumors. Co-downregulation of PTEN, KAI-1, and nm23-H1 significantly correlates with distant metastasis and predicts shortened survival. Our study supports a role of these tumor suppressor and metastasis suppressor genes in the evolution and progression of NSCLC. PMID- 15129905 TI - E-cadherin expression in invasive urothelial carcinoma. AB - E-cadherin (E-CD) is a transmembrane glycoprotein involved in intercellular adhesion. A loss or reduction in E-CD expression has been linked to the invasive phenotype of a wide variety of human neoplasms, including bladder tumors. The objective of this study was to compare the E-CD expression at different depths of tumor invasion below the bladder's basement membrane in high- and low-grade urothelial carcinomas to investigate whether deeper tumor invasion and higher grade invasive urothelial carcinomas are associated with decreased E-CD expression. E-cadherin staining was performed on 29 formalin-fixed, paraffin embedded sections from high- and low-grade urothelial carcinoma specimens using an automatic immunohistochemical stainer. The sections were divided into three categories according to the depth of invasion below the basement membrane: upper, middle, and lower. The percentage and intensity of E-CD cell membrane staining for the three categories were calculated using a quantitative automated cellular imaging system. The percentage of cells that stained for E-CD was 82.6% +/- 1.4% (mean +/- SD) in the upper layer, 59.6% +/- 2.2% in the middle layer, and 29.4% +/- 2.7% in the lower layer. The intensity of E-CD expression was 64.7 +/- 3.2 units in the upper layer, 43.3 +/- 2.9 units in the middle layer, and 26.1 +/- 3.1 units in the lower layer. There were significant differences between the three layers in both the percentage and intensity of cellular E-CD staining (P<.05). Normal urothelium, high-grade urothelial dysplasia/carcinoma in situ, and superficial noninvasive papillary urothelial carcinoma maintained E-CD expression. However, once malignant cells infiltrated through the basement membrane, E-CD expression decreased. The more poorly differentiated urothelial carcinoma, the deeper the nests, and the smaller the clusters of neoplastic cells within the tumor were, and the more decrease in E-CD expression noted. The degree of decreased E-CD expression was directly proportional to the degree of tumor differentiation and depth of infiltration in invasive urothelial carcinoma. Down regulation of E-CD may be one of the pathways responsible for tumor differentiation and may promote deeper invasion in urothelial carcinomas. PMID- 15129906 TI - Interobserver agreement for estrogen receptor immunohistochemical analysis in breast cancer: a comparison of manual and computer-assisted scoring methods. AB - Differences in scoring methods for estrogen receptor (ER) immunohistochemistry may cause significant variation in the results. Scoring practices differ within the United States and internationally and include semiquantitative scoring formulas, manual estimations, and computer-assisted techniques. The goal of this study was to determine the rate of interobserver variability for manual ER scoring at our institution and compare the ER scores obtained by manual scoring with those obtained using image-analysis software (QCA, Lake Bluff, IL). In a series of 70 consecutive invasive breast cancers, ER was assayed using standard immunohistochemical techniques and the monoclonal antibody 6F11. Scoring was performed independently by three breast pathologists, and the scores were compared with those obtained using the QCA image-analysis system, using 10% nuclear staining as the cutoff for positivity. We found that 43 cases (61%) were ER positive, 25 cases (36%) were ER negative, and two cases (3%) showed ER staining of less than 10%. The consensus scores for the 70 cases showed a high level of agreement with the ER scores determined by image analysis (kappa = 0.84). Interobserver variability was low. The kappa scores for each observer showed strong agreement with the consensus score, the image-analysis score, and between the observers. Our findings show that interobserver agreement for manual scoring of ER is strong, and that manual or computer-aided scoring techniques are comparable. PMID- 15129907 TI - Mature teratoma arising from the middle cranial fossa. AB - A 25-year-old man presented with a sudden spurt in size of a bony swelling around the external ear, which was present since birth. This was associated with a painful swelling of the right half of the face and pain, redness, and decreased vision in the right eye. Neurologic examination showed decreased visual acuity in the right eye and paresis of the fifth, seventh, and eighth cranial nerves. Neuroimaging showed an extra-axial, partly cystic tumor of mixed density, located entirely within the middle cranial fossa scalloping the temporal bone and extending into the right infratemporal fossa and the right middle ear and impinging on the ipsilateral cavernous sinus. The right temporal lobe was elevated and compressed. Histopathologic examination confirmed a mature teratoma. PMID- 15129908 TI - Gastric glomus tumor. AB - Gastric glomus tumors are rare neoplasms that may present with upper gastrointestinal symptoms, or may be discovered incidentally. They may mimic other tumors both grossly and microscopically. They are usually benign and, although the exceptional case with aggressive behavior cannot be reliably predicted, large size may be of prognostic importance. We describe a 3.8 cm gastric glomus tumor in a patient who also had adenocarcinoma arising in an adenoma of the rectum, and discuss the differential diagnosis and prognosis. PMID- 15129909 TI - Diffuse large B-cell lymphoma with anaplastic features and focal low-grade mucosa associated lymphoid tissue lymphoma component of the stomach. AB - We describe an 83-year-old man diagnosed as having diffuse, large B-cell lymphoma with anaplastic features and CD30 expression of the stomach. Focally, the tumor showed typical low-grade mucosa-associated lymphoid tissue lymphoma. We are not aware of a previous report on the transformation of low-grade gastric lymphoma of mucosa-associated lymphoid tissue type into an "anaplastic," large B-cell tumor. PMID- 15129910 TI - Invasive ductal carcinoma of the breast with a microglandular adenosis pattern. AB - Invasive breast carcinomas of special type are composed of up to 35% of all cases, such as tubular carcinoma, mucinous carcinoma, invasive cribriform carcinoma, lobular carcinoma, metaplastic carcinoma, medullary carcinoma, and other rare variants. They are recognized by their specific growth and morphologic patterns, glandular differentiation, and cytologic and nuclear features. These features are also present as a focal or minor component in many carcinomas of no special type. In addition, there are a number of lesions that are mimickers of invasive carcinoma, which can sometimes create diagnostic difficulties. It is important for pathologists to recognize these histologic variants and mimickers of breast carcinoma, particularly in evaluating needle or core biopsies. We report a case of a 64-year-old woman with breast carcinoma of no special type with areas having features with resemblance to microglandular adenosis. Differential diagnosis and their histologic features are discussed. PMID- 15129911 TI - Hepatocellular adenomatosis is a rare entity that may mimic other hepatocellular lesions. AB - A 14-year-old girl presented to her pediatrician with right lower quadrant pain that progressed to right upper quadrant pain with radiation to her back. Her past medical history included mitral valve prolapse, and she had no history of oral contraceptive use. Abdominal computed tomography revealed a liver mass and multiple smaller areas of low attenuation, too small to characterize. The clinical and radiographic features were suggestive of hepatocellular adenoma, and she underwent a left hepatic lobectomy. The liver contained one 4.2 cm nodule and multiple (10 to 20) smaller nodules that were well-demarcated from the adjacent liver parenchyma. All lesions were histologically hepatocellular adenomas and, therefore, she was diagnosed with hepatocellular adenomatosis. This case is unique because of the small number of cases of hepatocellular adenomatosis diagnosed in teenagers, and little long-term follow-up. PMID- 15129912 TI - Histologic and molecular assessment of human thymus. AB - Most work describing the histopathology of normal human thymus has focused on pediatric thymus because of tissue availability and high thymopoietic activity. However, pathologic examination of the thymus can provide information about immune status that is relevant to the clinical care of patients of all ages. Understanding age-related changes in the relative abundance and composition of anatomic compartments within the thymus is critical for evaluation of the thymus in normal adults and patients with diseases that affect the thymus. The purpose of this review is to acquaint diagnostic pathologists with some of the newer histologic, flow cytometric, and molecular techniques for assessment of non neoplastic thymus. Diagnostic criteria are presented for assessment of thymic function and for determining the mechanisms underlying thymic hyperplasia. Accurate assessment of thymic function is critical for the diagnosis and treatment of patients with complete DiGeorge syndrome and can complement the clinical care of patients with a variety of disorders that affect the immune system. PMID- 15129913 TI - An update on cystic ovarian degeneration in cattle. AB - Cystic ovarian degeneration (COD) is considered to be one of the most important causes of reproductive failure in cattle. There is a severe economic loss to dairy industry because COD increases days-open in the postpartum period and the culling rates. The disease process is a consequence of a mature follicle that fails to ovulate at the appointed time of ovulation in the oestrous cycle. This anovulatory follicular structure either regresses or persists as a follicular or luteal cyst depending upon its structural/functional characteristics. The cells lining the follicular cyst synthesize oestrogen that, in certain instances, forces the animal to exhibit clinical signs of nymphomania. Besides oestrogen production, as per recent findings, they are also capable of secreting varying amount of progesterone which may dictate their fate. The animals that carry a luteal cyst may tend to be in anoestrus as the higher amount of progesterone secreted by this luteinized structure may change the pattern of gonadotrophins' secretion. Present findings suggest that perturbation of the hypothalamo hypophyseal-ovarian (HHO) axis, due to many exogenous and endogenous factors, as the cause for anovulation. For example, it has been suggested that lack of hypothalamic or hypophyseal response to the positive feedback effect of oestrogens that are secreted by the dominant follicle as one of the many causes. The non-physiological changes that occur in the receptor expression of the HHO axis for the hormones involved in maturation, deviation, dominance and ovulation of the follicle may be yet another cause. The changes that occur at the cellular and molecular level in the ovary (in response to the factors mentioned above) that contribute to anovulation remain to be documented. This approach would allow us to completely understand the disease process. Hitherto, hormonal preparations that release luteinizing hormone from the anterior pituitary or have luteinizing hormone-like action are used to treat follicular cysts. GnRH belongs to the former group and human chorionic gonadotrophin (hCG) hormone forms the latter group. Treatment with a luteolytic agent, prostaglandin F2alpha (PGF2alpha), is successful if a luteal cyst is diagnosed properly. Many agents may be developed in the future if the cellular and molecular pathways of the disease process are delineated. This article will review recent advances in our understanding of the pathogenesis of COD and suggest direction for future studies to completely understand the disease mechanism. This review will also discuss the existing method of treatments for cysts and methods proposed for treatment of cysts that tend to be refractory in nature. PMID- 15129914 TI - Comparative study on five different commercial extenders for boar semen. AB - Increasing interest in a longer preservation of diluted boar sperm raises questions in the field concerning the choice of the extender. The aim of this study was to evaluate the longevity of boar sperm extended in currently used commercial semen extenders. Three long-term extenders and two short-term extenders were compared for different semen quality parameters that can be assessed under routine laboratory conditions. Sperm morphology, motility, pH and bacteriological contamination were investigated during a 7-day period. The number of dead spermatozoa did not differ significantly among the extenders (p > 0.05). Sperm motility was not only related with storage period but most of all with pH, especially in long-term extenders. Differences between the different extenders were prominent (p < 0.05); the sperm preserved in only one long-term extender showed good motility during the whole test period. In all cases, the pH of the extended semen increased by 0.3-0.5 in the first days of storage and was significantly correlated with a decrease in motility. Bacteriological quality had no significant influence on motility or pH of the semen. In conclusion, we can state that in both short-term extenders and in only one long-term extender, sperm longevity, as evaluated by the parameters used in this study, was sufficient during the preservation period. To preserve the quality of diluted boar semen during long-term storage, the choice of the long-term extender is important. In addition, the monitoring of the pH of extended boar semen in our study emphasizes the importance of the buffering capacity of semen extenders. PMID- 15129915 TI - Stage-specific expression of leukaemia inhibitory factor and its receptor in rabbit pre-implantation embryo and uterine epithelium during early pregnancy. AB - Leukaemia inhibitory factor (LIF) has been shown to play an important role in the development and implantation of blastocysts in mice. In the current study, the reverse transcription-polymerase chain reaction (RT-PCR) was employed to examine the expression patterns of LIF and its receptor (LIFR) genes in rabbit embryos during pre-implantation development, and the uterine expression of LIF and LIFR was also evaluated by Western blotting. Transcripts for LIFR were detected within morula and blastocyst-stage embryos, while the LIF mRNA was only found in blastocysts (from early to fully expanded blastocoel cavities), indicating that embryo-derived LIF can act in an autocrine manner on the process of blastocyst formation. The expression levels of LIF and LIFR in uterine epithelium were gradually increased during pre-implantation period and reached their highest levels on days 6.5 of pregnancy, just before the time of blastocyst implantation, suggest that paracrine LIF circuit should exist between the endometrium and the early embryos, which may be involved in the embryo-maternal dialogue and important for the blastocyst implantation. The data present here show the stage specific and dynamic expression patterns of LIF and LIFR, both in embryos and endometrium, during early pregnancy in rabbits, which indicated that LIF might play an important role in the pre-implantation development and subsequent implantation of rabbit embryos. PMID- 15129916 TI - Stored of Hsp72/Hsp73 in germinal vesicle-stage mouse oocytes. AB - Heat-shock proteins (hsps) Hsp72 and Hsp73 are the stored maternal proteins found in mouse oocytes. Both hsps appear in mouse oocytes at germinal vesicle (GV) and metaphase II (M-II)-stages as previously demonstrated by immunoblotting analysis. In this report, we further determined the presences of Hsp72/Hsp73 proteins in mouse embryos at stages of 2-pronucleus, arrested 1-cell, 2-cell, arrested 2 cell, 4-cell, arrested 4-cell, 8-cell to morula and blastocyst. Except for the blastocyst stage, the Hsp72/Hsp73 proteins were detectable in most embryo stages. The concentration of Hsp72/Hsp73 in GV-stage oocytes was higher than that in M-II stage oocytes, and in any stages of embryos before implantation. A dramatical increase in Hsp72/Hsp73 expression was found at the 2-cell stage. Together with these findings, we speculated that hsps accumulated or stored earlier in the GV stage mouse oocytes to protect the oocytes against environmental influences acting on ovary, and hsps may be required for zygotic gene activation and provided a protective effect against apoptosis. PMID- 15129917 TI - The influence of opioid peptides on steroidogenesis in porcine granulosa cells. AB - The present studies were undertaken to examine the influence of mu (beta endorphin, DAMGO, FK 33-824), delta (met-enkephalin, leu-enkephalin, DPLPE) and kappa opioid receptor agonists (dynorphin A, dynorphin B, U 50488) used at different doses (1-1000 nM) alone and in combination with LH (100 ng/ml) on steroidogenesis in porcine granulosa cells derived from large follicles. The effects of mu, delta and kappa receptor agonists on both basal and LH-induced progesterone (P4) secretion were negligible. Agonists of mu opioid receptors reduced basal androstenedione (A4), testosterone (T) and oestradiol (E2) release. Co-treatment with LH entirely abolished the inhibitory effect of these agonists on A4 and E2 secretion and resulted in an increase in T release. The addition of delta receptor agonists was followed by a decrease in basal A4, T and E2 secretion. The cells incubated in the presence of LH increased the androgen production and abrogated the inhibitory effect of delta agonists on E2 output. Basal A4, T and E2 release was also suppressed by kappa receptor agonists. The presence of LH in culture media extended the inhibitory effect of these opioids on E2 output and caused either abolition of the inhibitory influence of kappa agonists or even augmentation of both androgen release in response to the opioids. In conclusion, these data support the involvement of three major types of opioid receptors in the regulation of porcine granulosa cell steroidogenesis. PMID- 15129918 TI - In vitro development of buffalo oocytes in media-containing fluids from different size class follicles. AB - Studies were conducted to investigate the effect of supplementation of fluid from different sized class [small (SFF, < 3 mm), medium (MFF, 3-8 mm) and large (LFF, > 8 mm)] of normal and cystic (CFF) ovarian follicles in oocyte culture media on oocyte maturation rate and embryo development in vitro and to test the efficacy of follicular fluid (FF) from different size classes as a whole oocyte maturation medium. Results suggested that FF were capable of developing buffalo oocytes to embryonic stage in vitro although its efficacy was lower than that of serum. Regardless of high maturation rates after in vitro maturation (IVM) in media containing FF or IVM in whole FF, low blastocyst rates were obtained after in vitro fertilization (IVF) and culture of embryos. Follicular fluid from small follicles had significantly (p < 0.05) higher potential of developing buffalo oocytes to embryonic stage in vitro than that from medium and large follicles. Cystic FF was not capable of supporting development of buffalo oocytes in vitro. PMID- 15129919 TI - Poly(ADP-ribose) glycohydrolase in bovine retained and not retained placenta. AB - Poly(ADP-ribose) glycohydrolase (PARG) is the enzyme which degrades poly(ADP ribose) polymers synthesized by poly(ADP-ribose) polymerase (PARP). Both enzymes are activated in response to different stimuli like oxidative stress and are involved in DNA repair processes. The retention of bovine foetal membranes (RFM) is supposed to be connected with oxidative stress conditions. The aim of the study was to detect the presence of PARG protein in bovine placenta in order to find the relationship between the process of releasing, retaining placenta and DNA repair. Placentomes, collected alter spontaneous delivery or caesarian section were divided into maternal as well as foetal part of placenta, homogenized and subjected to electrophoresis. Animals were divided into six groups as follows: A--caesarian section before term with RFM; B--caesarian section before term without RFM; C--spontaneous delivery at term with RFM; D- spontaneous delivery at term without RFM; E--caesarian section at term with RFM; F--caesarian section at term without RFM. PARG protein was detected in nitrocellulose membranes using commercially available bovine anti-PARG antibody and Western blotting technique. Single bands referred to bovine PARG standard were observed in all examined tissues as well as in human placenta used as the control of procedure. In addition, the intensity of staining was stronger in retained than properly released term placenta and in foetal than in maternal part of the placenta. These results may suggest the differences in enzyme protein content and careful conclusions can be drawn that the activities of PARG may be altered between compared groups of animals. It may confirm the presence of oxidative stress conditions and their consequences on metabolic pathways, the content of biologically active substances and processes of proper releasing placenta. Further experiments on PARG activity in bovine foetal membranes with respect to proper and improper placental release are necessary. PMID- 15129920 TI - Expression of occludin in canine testis and epididymis. AB - Tight junctions (TJ) in inter-Sertoli junctional areas and epididymal epithelia are important for the formation of blood-testis barrier (BTB) and blood epididymal barrier (BEB). In this study, the expression of occludin, an integral member of the TJ, was verified in canine testis and epididymis. Both low molecular weight (MW) (25-28 kDa) forms as well as high MW (68-72 kDa) forms of occludin were detected in the testis and epididymis using Western blot. The relative amount of the high MW forms of occludin vs low MW forms was higher in the testis than in the epididymis. Some difference in the composition of different MW forms of occludin was found along the segments of epididymis, suggesting the possible correlation between cellular composition of occludin proteins and paracellular permeability of epithelia along the epididymal tubule. In the testis, intense occludin immunoreactivity was found in the basally located inter-Sertoli junctional area. Diffused immunoreactivity of occludin was also found in the cytoplasm of Sertoli cells. A similar pattern of zonula occludens-1 immunoreactivity was found in the cytoplasm of Sertoli cells, suggesting that occludin was not confined to the inter-Sertoli junctional areas and that subcellular localization of occludin in the Sertoli cells was dynamically regulated during spermatogenesis in canine testis. In the epididymis weak immunoreactivity was found in the apical sides and cytoplasm of epithelial cells. PMID- 15129921 TI - Pregnancy diagnosis in miniature pig by direct ELISA of oestrone derivatives in faeces. AB - The objectives of the present study were to measure oestrone derivatives [oestrone, oestrone sulphate (E1S) and oestrone glucuronide] in sow faeces by direct enzyme-linked immunosorbent assay (ELISA) and to explore the changes of oestrone derivative profile in faeces of miniature pig for demonstrating the possibility of pregnancy diagnosis. Faecal samples (1.5 g) were put into 6 ml of buffer, shaken and centrifuged. Then supernatant was added to the wells of multi plate without extraction and incubated with anti-oestrone antibody and horseradish peroxidase-labelled oestrone. Standard solution was prepared at various concentrations of E1S. Sensitivity was estimated as 0.035 ng/ml (0.14 ng/g). Intra- and inter-assay coefficient variations were 3.5-7.7% and 10.9 15.3%, respectively. When 1-5 ng/ml E1S were added to a faecal solution, recovery rates ranged between 80.0 and 103.3%. There is a temporal increase in the E1S equivalent concentration of miniature pig faeces from day 25 to 31 after mating. From day 35 to 70, the E1S equivalent concentrations remained low. Thereafter its concentrations increased again towards farrowing. On day 27 and 29 after mating pregnancy diagnosis by the faecal E1S equivalent concentrations agreed with the results of farrowing (seven of seven animals). These results suggest that the present direct ELISA is practical and suitable as a routine assay for measuring the faecal concentration of oestrone derivatives and that this assay might be usable for pregnancy diagnosis in sows at day 27-29. PMID- 15129922 TI - Exposure to L-ascorbic acid or alpha-tocopherol facilitates the development of porcine denuded oocytes from metaphase I to metaphase II and prevents cumulus cells from fragmentation. AB - It is known that alpha-tocopherol (vitamin E) and L-ascorbic acid (vitamin C) can modulate many biochemical processes intracellularly or extracellularly as antioxidants. The objective of the present study was to investigate the effects of alpha-tocopherol and L-ascorbic acid on porcine oocyte meiotic maturation, viability and the functions of cumulus cells. In two independent experiments, porcine oocytes with or free from cumulus cells were exposed to different levels of alpha-tocopherol (0, 10, 100 and 200 microM) or L-ascorbic acid (0, 50, 250 and 750 microM). Cumulus expansion, cumulus cell DNA fragmentation, meiotic maturation and degeneration of oocytes were assessed 48 h after in vitro culture. The results showed that: (1) neither alpha-tocopherol nor L-ascorbic acid influenced cumulus expansion but both prevented cumulus cell DNA fragmentation. (2) Alpha-tocopherol lowered the percentage of denuded oocytes (DOs) arrested at germinal vesicle stage (GV). Among the oocytes undergoing germinal vesicle breakdown (GVBD) proportion, fewer DOs treated by alpha-tocopherol were at metaphase I (MI) and more at metaphase II (MII). L-ascorbic acid caused lower percentage of DOs arrested at GV stage and higher percentage of DOs undergoing GVBD, especially at MII. The influences of alpha-tocopherol and L-ascorbic acid were not obvious in cumulus-enclosed oocytes (CEOs). (3) Both vitamins compromised the viability of CEOs and DOs. These results indicate that exposure to alpha-tocopherol or L-ascorbic acid promotes the development of porcine DOs from MI to MII and prevents cumulus cell DNA fragmentation at certain levels, especially 10 microM alpha-tocopherol or 250 microM L-ascorbic acid. PMID- 15129923 TI - Flow cytometry identification of X- and Y-chromosome-bearing goat spermatozoa. AB - Flow cytometric sorting technology was used to measure the difference in DNA content between X- and Y-chromosome-bearing spermatozoa in bucks. Spermatozoa were analysed by flow cytometry to characterize X- and Y-chromosome-bearing sperm populations and to quantify the DNA difference between them. Two symmetrical, overlapping and clearly separated peaks, corresponding to X- and Y-bearing spermatozoa, were detected. The difference in fluorescence intensity between the peaks was 4.4 +/- 0.03% without any significant inter- or intra-animal variations. Therefore, the identification and selection of high-purity samples of sperm populations for sex sorting is easier in bucks compared with other domestic species. PMID- 15129924 TI - Benign fibrous histiocytoma of prepuce in the mule. AB - Here we present a case of preputial benign fibrous histiocytoma in a 7-year-old mule. This case was treated surgically. Pathologic analysis and diagnosis of these lesions were based on a combination of light microscopy and immunohistochemistry. Histologically, the tumour was composed primarily of a storiform pattern of fibroblastic and poligonal cells. In immunohistochemistry, except for consistent immunoreactivity for vimentin, nearly all tumour cells were positive for CD68, S-100 protein, pan-cytokeratin, alpha-smooth muscle actin, and desmin. Benign fibrous histiocytoma of the prepuce in mule is rarely reported in veterinary literature. PMID- 15129925 TI - High-fat diets and performance: a response to Roltsch et al.'s "The effect of diet manipulations on aerobic performance". PMID- 15129926 TI - One year of oral calcium supplementation maintains cortical bone density in young adult female distance runners. AB - We conducted a double-blind, placebo-controlled, randomized trial to determine whether 1 year of supplemental calcium intake would augment hip [greater trochanter, GT, femoral neck (FN), total hip (TH)], spine (LS), and femoral mid shaft (Fmr) BMD in female distance runners. Twenty-three women (age: 23.7 +/- 4.7 yrs, height: 165.6 +/- 6.3 cm, weight: 55.7 +/- 6.1 kg) were randomly assigned to receive either 1000 mg/d of supplemental calcium (N = 13) or placebo tablets (N = 10) for 1 year. BMD was determined by DXA (Hologic 1000-W) and tablet compliance by self-report logs. Compliance averaged 79% and 71% for supplement and placebo groups, respectively. Calcium supplementation did not affect hip or spine BMD, but did prevent loss at the femoral mid-shaft (GT: -0.5% vs. 0.2%, FN: 0.9% vs. 1.1%, TH: -0.3% vs. 0.2%, LS: 0.3% vs. 1.2%, Fmr: 0.1% vs. -1.8%, for calcium vs. placebo, respectively). We conclude that the addition of 800 mg/d of supplemental calcium to the diet of young adult female distance runners with habitual calcium intakes of approximately 1000 mg/d, prevents cortical but not trabecular bone loss. PMID- 15129927 TI - A pilot intervention to increase calcium intake in female collegiate athletes. AB - Calcium intake in adolescent and young adult female athletes often is inadequate to optimize peak bone mass, an important determinant of osteoporosis risk. The purpose of this study was to determine if calcium supplementation in eumenorrheic female collegiate athletes increases intake to recommended levels and promotes increases in bone mineral density (BMD). Forty-eight eumenorrheic female athletes from several college teams (15 soccer, 7 cross-country, 8 indoor track, and 18 basketball) were randomized at the beginning of a competitive season to receive either an oral calcium supplement (1000 mg calcium citrate/400 I.U. Vitamin D) or placebo daily throughout the training season (16 weeks). Self-reported daily pill intake was obtained every 2 weeks to assess adherence. Calcium intake was evaluated using the Rapid Assessment Method, and total body and leg BMD was measured at pre-, mid-, and postseason using dual energy x-ray absorptiometry (DEXA; Hologic QDR-2000). Pre-season calcium intake was lower than national recommendations for this age group (12), averaging 842 mg/d (SD = 719) and was lower in the placebo group compared to the supplemented group (649 +/- 268 vs. 1071 +/- 986 mg/d, respectively; p = .064). Adherence to supplementation was good, averaging 70% across the training season. Supplementation boosted total calcium intake to a mean of 1397 +/- 411 mg/d, which is consistent with recommended levels for this group (37). Supplementation did not influence BMD change during this 16-week intervention. Across teams, a small increase of 0.8% was observed in leg BMD. Change in total body BMD was modified by team, with a significant increase of 1.5% observed in basketball players. These results indicate that providing calcium supplements of 1000 mg/d is adequate to boost total intake to recommended levels during athletic training. Longer intervention trials are required to determine whether calcium supplementation has a positive effect on BMD. PMID- 15129928 TI - Prevalence of iron depletion and anemia in top-level basketball players. AB - Iron depletion, with or without anemia, may have a negative effect on physical and mental performance. Even with current recognition of the problem, its incidence among athletes remains high. Most studies describe iron status in endurance athletes. This study examined the prevalence of iron depletion and anemia among male and female top-level basketball players. Adolescents and adults (N = 103) from 8 national basketball teams were screened for anemia and iron stores status, which included a complete blood count and levels of plasma ferritin, transferrin, and serum iron. Iron depletion, defined by a ferritin level below 20 microg/L, was found among 22% of study participants (15% in males vs. 35% in females, p = .019). Anemia was found among 25% of athletes (18% in males vs. 38% in females, p = .028). Iron deficiency anemia, defined by the presence of anemia, ferritin levels below 12 microg/L, and transferrin saturation below 16%, was found among 7% of players (3% in males vs. 14% in females, p = .043). In summary, a high prevalence of iron depletion, anemia, and iron deficiency anemia was found among basketball players of both genders. We recommend screening ballgame players for blood count and iron store status, and providing nutritional counseling and iron supplementation when necessary. PMID- 15129929 TI - Dietary composition influences short-term endurance training-induced adaptations of substrate partitioning during exercise. AB - The purpose of the current study was to examine the influence of dietary composition on short-term endurance training-induced adaptations of substrate partitioning and time trial exercise performance. Eight untrained males cycled for 90 min at approximately 54% aerobic capacity while being infused with [6,6(2)H]glucose before and after two 10-d experimental phases separated by a 2 week washout period. Time trial performance was measured after the 90-min exercise trials before and after the 2nd experimental phase. During the first 10 d phase, subjects were randomly assigned to consume either a high carbohydrate or high fat diet while remaining inactive (CHO or FAT). During the second 10-d phase, subjects consumed the opposite diet, and both groups performed identical daily supervised endurance training (CHO+T or FAT+T). CHO and CHO+T did not affect exercise metabolism. FAT reduced glucose flux at the end of exercise, while FAT+T substantially increased whole body lipid oxidation during exercise and reduced glucose flux at the end of exercise. Despite these differences in adaptation of substrate use, training resulted in similar improvements in time trial performance for both groups. We conclude that (a) 10-d high fat diets result in substantial increases in whole body lipid oxidation during exercise when combined with daily aerobic training, and (b) diet does not affect short term training-induced improvements in high-intensity time trial performance. PMID- 15129930 TI - The influence of low versus high carbohydrate diet on a 45-min strenuous cycling exercise. AB - To examine the effects of a 3-day high carbohydrate (H-CHO) and low carbohydrate (L-CHO) diet on 45 min of cycling exercise, 12 endurance-trained cyclists performed a 45-min cycling exercise at 82 +/- 2% VO2peak following an overnight fast, after a 6-day diet and exercise control. The 7-day protocol was repeated under 2 randomly assigned dietary trials H-CHO and L-CHO. On days 1-3, subjects consumed a mixed diet for both trials and for days 4-6 consumed isocaloric diets that contained either 600 g or 100 g of carbohydrates, for the H-CHO and the L CHO trials, respectively. Muscle biopsy samples, taken from the vastus lateralis prior to the beginning of the 45-min cycling test, indicated that muscle glycogen levels were significantly higher (p < .05) for the H-CHO trial (104.5 +/- 9.4 mmol/kg wet wt) when compared to the L-CHO trial (72.2 +/- 5.6 mmol/kg wet wt). Heart rate, ratings of perceived exertion, oxygen uptake, and respiratory quotient during exercise were not significantly different between the 2 trials. Serum glucose during exercise for the H-CHO trial significantly increased (p < .05) from 4.5 +/- 0.1 mmol x L(-1) (pre) to 6.7 +/- 0.6 mmol x L(-1) (post), while no changes were found for the L-CHO trial. In addition, post-exercise serum glucose was significantly greater (p < .05) for the H-CHO trial when compared to the L-CHO trial (H-CHO, 6.7 +/- 0.6 mmol x L(-1); L-CHO, 5.2 +/- 0.2 mmol x L( 1)). No significant changes were observed in serum free fatty acid, triglycerides, or insulin concentration in either trial. The findings suggest that L-CHO had no major effect on 45-min cycling exercise that was not observed with H-CHO when the total energy intake was adequate. PMID- 15129931 TI - Metabolic effects of the addition of resistive to aerobic exercise in older men. AB - The metabolic changes associated with the addition of 4 months of resistive exercise to an existing aerobic exercise program (AEX+RT, n = 7) were compared to a maintenance aerobic exercise program (AEX, n = 8) in overweight, older men. The subjects in this study had recently completed a 6-month aerobic exercise program (treadmill walking, 45 min/d, 2 d/wk). The AEX+RT group added 6 exercises on upper- and lower-body pneumatic-resistance machines (2 sets, 15 repetitions each, 2 d/wk) to an aerobic exercise program at > or = 70% heart rate reserve for 30-40 min, 2 d/wk on treadmill, while the AEX group continued the same maintenance treadmill AEX program. There were no baseline differences in body weight, VO2max, or glucose metabolism between groups. The AEX+RT group increased upper- and lower extremity strength by 28 +/- 4% and 46 +/- 6%, respectively (p < .05), despite a 9% decrease in VO2max (p < .05). VO2max did not change in the AEX group. There was no change in the fasting glucose or insulin levels, or the 3-h glucose responses to an oral glucose load in either group. The insulin responses decreased by 25 +/- 4% in the AEX+RT group (p < .01) but did not change in the AEX group. In conclusion, the addition of resistive exercise training to an existing aerobic exercise program may improve insulin sensitivity in overweight, older men, and thus prevent the development of type 2 diabetes. PMID- 15129932 TI - Nutritional status of Brazilian elite swimmers. AB - The purpose of this study was to evaluate the body composition, dietary intake, use of nutritional supplements, and biochemical status of 8 Brazilian male elite swimmers, aged 18-21 years, participants at a national swimming competition. Data from the athletes were obtained through a 4-day food record, a fasting blood sample, and anthropometric measurements. The anthropometric results showed that body composition was compatible with sport category. The dietary assessment showed an adequate ingestion of calories, vitamins, and mineral, with the exception of calcium, for which only half of the sample reached the recommendation. The results also indicated low carbohydrate and high protein and cholesterol intakes. Of the swimmers, 62.5% and 25% consumed synthetic aminoacids and antioxidants supplements, respectively. The biochemical indices of the nutritional status were within normal limits in all swimmers, with the exception of creatine-kinase, which was above the recommended level, indicating muscle degradation probably due to poor carbohydrate intake. In conclusion, the results suggest the importance of nutritional education to promote a balanced intake, provide all nutrients in optimal amounts, inhibit unnecessary ingestion of nutritional supplements, maintain ideal performance, and improve the swimmers' health status. PMID- 15129933 TI - Creatine supplementation in young soccer players. AB - The purpose of this study was to examine the effects of acute creatine monohydrate supplementation on soccer-specific performance in young soccer players. Twenty young male soccer players (16.6 +/- 1.9 years) participated in the study and were matched and allocated to 2 randomly assigned trials: ingesting creatine-monohydrate supplement (3 x 10-g doses) or placebo for 7 days. Before and after the supplementation protocol, each subject underwent a series of soccer specific skill tests: dribble test, sprint-power test, endurance test, and vertical jump test. Specific dribble test times improved significantly in the creatine group (13.0 +/- 1.5 vs. 10.2 +/- 1.8 s; p < .05) after supplementation protocol. Sprint-power test times were significantly improved after creatine monohydrate supplementation (2.7 +/- 0.4 vs. 2.2 +/- 0.5 s; p < .05) as well as vertical jump height (49.2 +/- 5.9 vs. 55.1 +/- 6.3 cm; p < .05) in creatine trial. Furthermore, dribble and power test times, along with vertical jump height, were superior in creatine versus placebo trial (p < .05) at post supplementation performance. There were no changes in specific endurance test results within or between trials (p > .05). There were no between-trial differences in the placebo trial (p > .05). The main finding of the present study indicates that supplementation with creatine in young soccer players improved soccer-specific skill performance compared with ingestion of placebo. PMID- 15129934 TI - Nutritional supplement use among college athletes and their sources of information. AB - A survey was conducted to examine the source of information and usage of nutritional supplements in 115 male and 88 female varsity athletes at a Division I university. The survey asked each athlete to define supplement, and report supplement use and type, source of information, and reasons for use. Supplement use frequencies were determined, and comparisons were made between gender and sport. Eighty-nine percent of the subjects had or were currently using nutritional supplements. Many athletes did not consider sports drinks and calorie replacement products as supplements. Females were more likely to take calcium and multivitamins, and males had significant intake for ginseng, amino acids, glutamine, hydroxy-methyl-buterate (HMB), weight gainers, whey protein, and Juven. The most frequently used supplements overall were energy drinks (73%), calorie replacement products of all types (61.4%), multivitamin (47.3%), creatine (37.2%), and vitamin C (32.4%). There was also significant supplement use noted per sport. Females were more likely to obtain information from family members regarding supplementation, and males from a store nutritionist, fellow athletes, friends, or a coach. Female athletes were more likely to take supplements for their health or because of an inadequate diet, while men reported taking supplements to improve speed and agility, strength and power, or for weight/muscle gain. PMID- 15129935 TI - National Clostridium difficile Standards Group: Report to the Department of Health. PMID- 15129936 TI - Chronic esophageal foreign bodies in pediatric patients: a retrospective review. AB - OBJECTIVE: Chronic esophageal foreign bodies (CEFB) are associated with a high incidence of morbidity and mortality in adults. However, the presentation, management and outcome of chronic esophageal foreign bodies in children are not well described. METHODS: We performed a retrospective chart review of children with chronic esophageal foreign bodies admitted to the Children's Hospital Medical Center, Cincinnati, OH, between May 1990 and January 2002. A chronic esophageal foreign body was defined as a foreign body estimated to have been present for over 1 week. RESULTS: Over the inclusion period, 522 children were admitted with esophageal foreign bodies, 41 (8%) of which were chronic. The most common foreign bodies were coins. Seventy-six percent of patients presented with a primary complaint of respiratory symptoms, with respiratory distress being the most common followed by asthmatic symptoms and cough. Twenty-two percent of patients had primarily gastrointestinal symptoms including nausea/vomiting and dysphagia. One patient was asymptomatic on presentation. A perforated esophagus was identified in 18 patients, with 17 of these being a technically perforated esophagus and one case being a classic esophageal perforation. There were no deaths or permanent morbidity in this series. CONCLUSIONS: Respiratory symptoms are more common than gastrointestinal symptoms in pediatric patients with chronic esophageal foreign bodies. Removal by rigid esophagoscopy is recommended. A small proportion of cases require open removal of the foreign body. Conservative management is appropriate for the technically perforated esophagus. A good outcome should be anticipated for the majority of cases. PMID- 15129937 TI - Cochlear implantation in children with otitis media: second stage of a long-term prospective study. AB - OBJECTIVE: To present the findings of the second stage of an ongoing prospective study of the outcome of cochlear implantation in otitis media (OM)-prone and non OM-prone children who were treated according to a structured protocol designed to control OM prior to implantation. PATIENTS AND METHODS: Of 60 children referred for cochlear implantation during the study period, 34 were classified as OM-prone (Group A) and 26 as non-OM-prone (group B). Group A patients were managed according to a structured protocol aimed at pre-implantation control of OM. A ventilating tube (with or without adenoidectomy) was inserted in the affected ear(s) of these children, if necessary more than once. Post-implantation follow up ranged from 3 to 45 months (average 20 months). RESULTS: In the OM-prone group of children, the mean age at referral and at implantation was significantly lower and the mean interval between referral and implantation significantly higher than in the healthy group. During implantation, 21 children (19 from the OM-prone group A) had thick middle ear mucosa that had to be removed to allow identification of the round window niche. After implantation, 13 of the OM-prone children (38%) and 2 of the non-OM-prone children (7.6%) developed acute OM in the implanted ear. Five of these cases, all belonging to the OM-prone group, proved to be recurrent and therapeutically challenging, and 2 of them also developed acute mastoiditis. There were no other OM-related complications. CONCLUSIONS: Early referral led to early implantation, even in children susceptible to OM. The incidence of OM decreased after implantation, but was still significantly higher in the OM-prone group. On the basis of these results, we recommend the continuous use of a ventilating tube in OM-prone pediatric implantees until they outgrow their susceptibility to OM. PMID- 15129938 TI - Is visual perception of hearing-impaired children different from healthy children? AB - OBJECTIVE: The purpose of this study was to evaluate visual perception of hearing impaired children, and to determine their insufficiency in rehabilitation programs. METHODS: Forty children with hearing impairment aged 8-10 years were evaluated, and were compared with age matched 40 healthy children. Children having 71 dB and over sensorineural auditory impairment in both ears were included in this study. Figure-ground perception, position in space, and design copying tests were used to evaluate the visual perception of the subjects (Ayres Southern California Sensorial Integration tests). RESULTS: The mean hearing impairment level was 95.5 +/- 13.86 dB for the right ear, and 92.25 +/- 14.3 dB for the left. There were no significant differences by mean of age, height, and body weight between the groups (P > 0.05). All of the test scores of the control group were significantly higher than those of hearing-impaired children (P < 0.05). However there was no significant difference in the completion time of the design copying test between the groups (P > 0.05). CONCLUSIONS: Motivation insufficiency and learning difficulty may be developed in hearing-impaired children depending on the communication problems. The result of this study may bring light into literature about the development of new assessment techniques, and proper rehabilitation programmes for hearing-impaired children or adults in different age groups. PMID- 15129939 TI - Benefits and challenges of newborn hearing screening for developing countries. AB - The late detection of permanent congenital and early-onset hearing loss (PCEHL) often has severe effects on linguistic, speech, cognitive and educational development in affected children. Since newborn hearing screening (NHS) allows most PCEHL to be detected early enough for optimal intervention, the prospects of its introduction in the developing world are reviewed in this paper. It is observed that a simple generalisation on the feasibility of NHS for the developing countries seems inappropriate in view of the diversities in the health and socio-economic status of these countries and the recent favourable reports of universal newborn hearing screening from the region. NHS empowers parents to make timely choices that will allow their hearing impaired children to be given a good start in life and be fully integrated into the wider community. It also compels attention towards the development of essential hearing healthcare services, besides the specific documented benefits. Existing child-healthcare structures such as the expanded programme on immunisation (EPI), baby friendly hospital initiatives (BFHI) and integrated management of childhood illness (IMCI) provide opportunities for the introduction of some form of NHS in many of these countries where routine or systematic childhood hearing screening does not exist. Limited funding, manpower shortages, inadequate support services, low public awareness and the uncertainty regarding the commitment from healthcare practitioners may present some challenges but these are not insurmountable. Pilot studies are necessary in each country to provide empirical data that will guide healthcare providers who wish to introduce such a programme at any level of healthcare delivery. PMID- 15129940 TI - Are tonsils a reservoir for Helicobacter pylori infection in children? AB - OBJECTIVE: Helicobacter pylori (H. Pylori) has been associated with the development of gastritis, peptic ulcer and gastric cancer. Although H. pylori infects up to more than half of the world's population, to date the precise modes of transmission has not been fully understood yet. Therefore a study was planned to investigate whether the tonsils and the adenoid tissue were the reservoir or the gate for the entrance of H. pylori. METHODS: The adenotonsillectomy specimens obtained from 50 children, between two and 10 years of age were examined for H. Pylori colonization by the CLO-test method. Before tonsillectomy, anti-H. pylori IgG antibody titers were detected by commercial enzyme immunoassay method in venous blood and H. pylori antigen enzyme immunoassay for detection of H. pylori in stool was used to define current infection status. RESULTS: The stool antigen was positive in 25 (50%) of 50 children. Serum IgG antibody was positive in 28 (56%) patients and both tests were positive in 21 (42%) patients. H. pylori positivity was not detected in any one of the adenotonsillectomy specimens with the CLO-test method. CONCLUSION: In this study although H. pylori was detected in stools of children, it was not detected in adenotonsillectomy specimens with CLO test method. The results may indicate that H. pylori does not colonize in either adenoid or tonsils and that these tissues do not constitute a reservoir for H. pylori infection. PMID- 15129941 TI - Pre- and postoperative evaluation by acoustic rhinometry of children submitted to adenoidectomy or adenotonsillectomy. AB - INTRODUCTION: Nasal obstruction is a common symptom in children and its etiology includes septal deviation, choanal atresia, allergic rhinitis and hypertrophy of the adenoids. Hypertrophy of the adenoids and hypertrophic rhinitis are the most frequent causes of nasal obstruction in the pediatric population, with adenoidectomy being the main surgery carried out during childhood. OBJECTIVE: To analyze the rhinograms of children with nasal obstruction before and after surgery and to compare them with those obtained for children without respiratory complaints. STUDY DESIGN: A clinical prospective study. METHODS: Thirty-five patients with adenoid or adenotonsillar hypertrophy were submitted to otolaryngologic examination and acoustic rhinometry before and 30-60 days after surgery. The control group consisted of 18 children without nasal complaints. RESULTS: Significant differences in the rhinograms were observed before and after surgery, but not between patients and the control group. CONCLUSION: We conclude that acoustic rhinometry is well tolerated by children, and is a rapid and noninvasive method. The technique is valuable for interindividual comparisons, but not for the assessment of different groups. PMID- 15129942 TI - Complications of acute otitis media in children in southern Finland. AB - BACKGROUND: The incidence of intratemporal and intracranial complications of acute otitis media (AOM) has decreased and the need for operative treatment is declined in developed countries during the antibiotic era. OBJECTIVES: To establish the clinical picture, diagnostic procedures, outcome and current treatment of pediatric patients with intratemporal and intracranial complications of AOM. METHODS: A retrospective chart review with a sent questionnaire. All pediatric patients treated for intratemporal and intracranial complications of AOM over the past 10 years (1990-2000) at the Department of Otolaryngology in the Helsinki University Central Hospital. RESULTS: During the study period 33 children (incidence 1.1/100,000 per year), aged from 3 months to 14.2 years were treated for intratemporal [97% (32/33)] and intracranial [3% (1/33)] complications of AOM. Facial paresis was found in 9% (3/33) of the patients. The only intracranial complication was an extradural abscess with meningitis. Eighteen patients (55%) were on antibiotic treatment because of AOM prior to the diagnosis of complication. Neither the duration or severity of the signs and symptoms of infection at the time of admittance nor a lack of antibiotic treatment before admittance were statistically significantly associated with the need for mastoidectomy or duration of hospitalization. Streptococcus pneumoniae 25% (8/33) and Pseudomonas aeruginosa 22% (7/33) were the most frequently found bacteria in the culture of middle ear and mastoid effusions. Mastoidectomy was performed on 55% (18/33) of the patients. After half a year of follow-up, all the patients had normal hearing and facial function. CONCLUSIONS: Severe complications of AOM are rare today in southern Finland and the need for mastoidectomy has declined significantly. With early recognition and effective treatment of complications, the prognosis is good. PMID- 15129943 TI - "Starplasty" prevents tracheotomy complications in infants. AB - OBJECTIVE: The average age of children undergoing tracheotomy has declined over the years and the indications for tracheotomy have changed from acute airway obstruction due to infection, to treatment of chronically ill children. Tracheotomy-related complication rates are more numerous in younger children and in certain cases have proved to be fatal. A variety of operative techniques have been advocated for reducing the rate of tracheotomy-related morbidity and mortality. This manuscript reports on our experience with a new technique- starplasty tracheotomy (SPT)--and assesses the value of this procedure in preventing tracheotomy-related complications in infants (under 6 months old), in whom complications are more frequent and more dangerous. METHODS: Children less than 6 months old who underwent tracheotomy in the Department of Otolaryngology/Head and Neck Surgery, Hadassah University Hospital, Jerusalem, between the years 1999 and 2003 were studied. They were divided into two groups according to the surgical technique preferred and performed by the senior surgeon on call. Children in group 1 underwent tracheotomy by means of a regular technique and children in group 2 underwent SPT. All events related to the tracheotomy were recorded and a comparison was made between the two groups. RESULTS: Eleven of a total of 26 pediatric tracheotomy patients were less than 6 months old. Five underwent regular tracheotomy (RT) and six underwent SPT. Three complications were encountered in the RT group and none in the SPT group. Accidental decannulation and failure to re-insert the cannula resulted in severe hypoxemic brain damage in one patient with RT. No tracheotomy-related deaths occurred. CONCLUSION: Our experience supports the existing data regarding the superiority of SPT over RT in preventing dangerous tracheotomy-related complications in infants. We, therefore, recommend performing SPT in all infants undergoing tracheotomy, even at the price of having to perform surgical closure of TCF after decannulation in some cases. PMID- 15129944 TI - Otitis media with effusion: disease or defense? A review of the literature. AB - Many studies of otitis media with effusion (OME) have been published, most of them dealing with risk factors. The literature correlates this condition with various patient characteristics and socio-economic factors, but none of these have been identified as the sole causative factor. A review of the literature suggests that otitis media with effusion is a response to pathogenic bacteria and thus a normal protective reaction of the body. Therefore, the decision on whether or not treatment is indicated should take the natural course of that response into account. In light of the literature reviewed here, we conclude that there is usually no need to treat middle ear effusion in young children. PMID- 15129945 TI - A novel laser-assisted stapedotomy technique for congenital stapes fixation. AB - OBJECTIVE: To investigate whether a novel laser-assisted stapedotomy technique is effective in treating congenital stapes fixation. METHODS: With this surgical method, the suprastructure of the stapes is vaporized continuously from the superior half of the anterior crus to the inferior part of the posterior crus using a potassium-titanyl-phosphate (KTP) laser. The midmost point of the vaporization is the head of the stapes. This method was used to treat three patients with congenital stapes fixation (total of four ears). RESULTS: The average hearing gain using this method was 32.2 dB, which lasted for at least 1 year. No complications were observed. CONCLUSIONS: This novel laser-assisted stapedotomy technique is an excellent method for the treatment of congenital stapes fixation. PMID- 15129946 TI - Speech perception and speech intelligibility in children after cochlear implantation. AB - OBJECTIVE: This study aimed to evaluate the long-term speech perception and speech intelligibility of congenitally and prelingually deaf children after cochlear implantation. It was a longitudinal study following 63 congenitally or prelingually deaf children up to 5 years after implantation. They each received a nucleus multichannel cochlear implant before they were 10 years old. METHODS: Perception is evaluated using the Test for the Evaluation of Voice Perception and Production (TEPP) and concerns closed- and open-set word and sentence perception without lip-reading. The intelligibility is classified according to the Speech Intelligibility Rating (SIR). The evaluations have been made every 3 months for 1 year, then at 18 months, 2 years, 3 years and 5 years after the cochlear implantation. RESULTS: After 5 years of implantation, the median percentage of closed-words speech perception (CSW) is 95.5%-93.67% for closed-sentence speech perception (CSS) and 76.3% for open-sentence speech perception (OSS); the median Speech Intelligibility Rating is 3.83. CONCLUSIONS: Congenitally and prelingually deaf children who receive cochlear implant before the age of 10 years develop speech perception and speech intelligibility abilities. The closed-set perception progresses quickly and seems to reaching a plateau at 5 years post implantation. The improvement of open-sentence perception is not significant until the first year post implantation. The speech intelligibility improves regularly the five first year post implantation. PMID- 15129947 TI - The carpenter syndrome phenotype. AB - Carpenter syndrome (Acrocephalopolysyndactyly type II), first described in 1901, consists of acrocephaly, syndactyly, polydactyly, congenital heart disease, mental retardation, hypogenitalism, cryptorchidism, obesity, umbilical hernia and bony abnormalities. We report a 6 years old boy presenting as a union of these malformations and also having bilateral sensorineural hearing loss. Auditory disturbances are not common among Carpenter syndrome patients. According to our knowledge, this is the first Carpenter syndrome case whose hearing loss is demonstrated by auditory brainstem response (ABR) test. PMID- 15129948 TI - Two unusual lesions in the nasal cavity of infants--a nasal chondromesenchymal hamartoma and an aneurysmal bone cyst like lesion. More closely related than we think? AB - Benign reparative lesions in the head and neck region in infants are rare and often difficult to classify on histology. Discussed herein are two rare lesions in infants occurring at identical locations in the nasal cavity with striking histologic similarity but different histologic labels. One was a case of nasal chondromesenchymal hamartoma (NCMH) occurring in a 1-year-old child and the other an aneurysmal bone cyst (ABC) like lesion affecting a 4-month infant. Both these lesions were locally destructive and had nearly similar clinical presentation. Both on immunohistochemistry showed myofibroblastic nature and had similar histology except that the ABC like lesion lacked the cartilage component of the former. In view of great similarity in the two lesions, it was thought that the second lesion might also represent a reparative, non-cartilage-containing counterpart of the former. PMID- 15129949 TI - Capillary haemangioma of the nasal cavity in a 7-week-old baby--successful treatment using intralesional steroid injection. AB - A case of a seven-week-old baby who presented with a capillary haemangioma of the nasal cavity is presented. We describe a novel treatment modality and provide a literature review of head and neck capillary haemangioma in the paediatric population. Capillary haemangioma of the nasal cavity and our treatment option has not previously been described in the literature at such an early age. PMID- 15129950 TI - Hydatid cyst of the thyroid gland in a child. AB - The hydatid cyst tend to form in the liver or lung but may be found in any organ of the body, including brain, heart, and bones. Thyroid gland involvement is rather rare. Cyst leakage or rupture may even associated with severe allergic reaction to parasite antigens, and may cause anaphylactoid reactions. Herein we reported an additional case of hydatid cyst of the thyroid gland that occurred in a 9-year-old boy who admitted with a painless and cystic thyroid mass. Hemithyroidectomy was performed, and histopathology confirmed the diagnosis of hydatid cyst. Diagnosis of that rare clinical entity is unlikely prior to surgical excision and histopathologic examination. Despite this fact, hydatid disease of the thyroid should be remembered in the differential diagnosis of the cystic thyroid masses, and a high index of suspicion is required. PMID- 15129951 TI - Pott's puffy tumour in a pre-adolescent child: the youngest reported in the post antibiotic era. AB - Pott's puffy tumour is a subperisosteal abscess associated with a cranial osteomyelitis. A rare consequence of trauma or frontal sinusitis, it is often an indicator of intracranial complications. At 3 years of age, the patient described is the youngest reported with this condition. The occurrence of such pathology in pre-adolescent children is very rare and this case is one of three reported in the English literature in the post-antibiotic era. PMID- 15129952 TI - A model of pollen-mediated gene flow for oilseed rape. AB - The development of genetically modified (GM) crops has precipitated the need for risk assessment and regulation of pollen-mediated gene flow. In response to this need we present a mathematical model to predict the spatial distribution of outcrossing between progenitor populations of oilseed rape. The model combines the processes of pollen dispersal and pollination, resulting from wind and insect activity. It includes the effects of post-pollination reproductive processes by relating the number of progeny to both pollen deposition and competition at the stigma. Predictions compare well with a range of experimental results for different-sized GM source crops (i.e. 0.0064-0.8 ha) and non-GM target crops with different fertilities (i.e. self-fertile to 80% male-sterile). For these comparisons, we represent the variation caused by wind and insect exposure as a constrained set of random functions and limit the range of insect transport to typical plant-scale distances. In addition, the model is used to examine the relative sensitivity to the factors that determine gene flow. Target-crop fertility and source-crop size are shown to be more important than other factors, including background pollen and the natural range of insect activity. The concept of isolation distance to regulate gene flow is most effective for self-fertile target crops, but is ineffective for male-sterile target crops with low background pollen. PMID- 15129953 TI - The selfish nature of generosity: harassment and food sharing in primates. AB - Animals may share food to gain immediate or delayed fitness benefits. Previous studies of sharing have concentrated on delayed benefits such as reciprocity, trade and punishment. This study tests an alternative model (the harassment or sharing-under-pressure hypothesis) in which a food owner immediately benefits because sharing avoids costly harassment from a beggar. I present an experiment that varies the potential ability of the beggar to harass, and of the owner to defend the food, to examine the effects of harassment on food sharing in two primate species: chimpanzees (Pan troglodytes) and squirrel monkeys (Saimiri boliviensis). For both species, high levels of harassment potential significantly increased both beggar harassment and sharing by the owner. Food defensibility did not affect harassment or sharing. Interestingly, squirrel monkeys and chimpanzees shared equally frequently with conspecifics despite a much higher natural sharing rate in chimpanzees. These results suggest that harassment can play a significant role in primate food sharing, providing a simple alternative to reciprocity. The selfish nature of harassment has implications for economic, psychological and evolutionary studies of cooperative systems. PMID- 15129954 TI - To age or not to age. AB - According to the antagonistic pleiotropy theory of ageing, natural selection has favoured genes conferring short-term benefits to the organism at the cost of deterioration in later life. The 'disposable soma' theory expresses this as a life-history strategy in which somatic maintenance is below the level required to prevent ageing, thus enabling higher immediate fertility. It has been argued that a non-ageing strategy will always be bettered by a low but non-zero rate of ageing, because the costs of such ageing will be felt only in the distant future when they are of negligible importance. Here, we examine this argument critically. We find that a non-ageing strategy will be locally optimal if, in the presence of ageing, the onset of deterioration is sufficiently rapid or early. Conversely, ageing will be optimal if deterioration is sufficiently slow or late. As the temporal profile of ageing changes from one of steady deterioration to one involving a sudden loss of vitality after a period of little or no decline, the conditions for a non-ageing strategy to be locally optimal become progressively more stringent. But for all forms of profile considered, conditions can be found for which a strategy involving no ageing is locally optimal. PMID- 15129955 TI - An evolutionary model of stature, age at first birth and reproductive success in Gambian women. AB - We have built a model to predict optimal age at first birth for women in a natural fertility population. The only existing fully evolutionary model, based on Ache hunter-gatherers, argues that as women gain weight, their fertility (rate of giving birth) increases-thus age at first birth represents a trade-off between time allocated to weight gain and greater fertility when mature. We identify the life-history implications of female age at first birth in a Gambian population, using uniquely detailed longitudinal data collected from 1950 to date. We use height rather than weight as an indicator of growth as it is more strongly correlated with age at first birth. Stature does not greatly influence fertility in this population but has a significant effect on offspring mortality. We model age at first reproduction as a trade-off between the time spent growing and reduced infant mortality after maturation. Parameters derived from this population are fitted to show that the predicted optimal mean age of first birth, which maximizes reproductive success, is 18 years, very close to that observed. The reaction norm associated with variation in growth rate during childhood also satisfactorily predicts the variation in age at first birth. PMID- 15129956 TI - Evolution of size-dependent flowering in a variable environment: partitioning the effects of fluctuating selection. AB - In a stochastic environment, two distinct processes, namely nonlinear averaging and non-equilibrium dynamics, influence fitness. We develop methods for decomposing the effects of temporal variation in demography into contributions from nonlinear averaging and non-equilibrium dynamics. We illustrate the approach using Carlina vulgaris, a monocarpic species in which recruitment, growth and survival all vary from year to year. In Carlina the absolute effect of temporal variation on the evolutionarily stable flowering strategy is substantial (ca. 50% of the evolutionarily stable flowering size) but the net effect is much smaller (ca. 10%) because the effects of temporal variation do not influence the evolutionarily stable strategy in the same direction. PMID- 15129957 TI - The mathematics of motion camouflage. AB - Motion camouflage is a strategy whereby an aggressor moves towards a target while appearing stationary to the target except for the inevitable change in perceived size of the aggressor as it approaches. The strategy has been observed in insects, and mathematical models using discrete time or neural-network control have been used to simulate the behaviour. Here, the differential equations for motion camouflage are derived and some simple cases are analysed. These equations are easy to simulate numerically, and simulations indicate that motion camouflage is more efficient than the classical pursuit strategy ('move directly towards the target'). PMID- 15129958 TI - Motor mechanisms of a vocal mimic: implications for birdsong production. AB - The diverse vocal performances of oscine songbirds are produced by the independent but coordinated patterns of activity in muscles controlling separate sound generators on the left and right sides of their duplex vocal organ, the syrinx. Species with different song styles use the two sides of their syrinx in different ways to produce their species-typical songs. Understanding how a vocal mimic copies another species' song may provide an insight into whether there are alternative motor mechanisms for generating the model's song and what parts of his song are most difficult to produce. We show here that when a vocal mimic, the northern mockingbird, accurately copies the song of another species it also uses the vocal motor pattern employed by the model species. Deviations from the model's production mechanism result in predictable differences in the mockingbird's song. Species-specific acoustic features of the model seem most difficult to copy, suggesting that they have been exposed to the strongest selective pressure to maximize their performance. PMID- 15129959 TI - Temporal stability of insular avian malarial parasite communities. AB - Avian malaria is caused by a diverse community of genetically differentiated parasites of the genera Plasmodium and Haemoproteus. Rapid seasonal and annual antigenic allele turnover resulting from selection by host immune systems, as observed in some parasite populations infecting humans, may extend analogously to dynamic species compositions within communities of avian malarial parasites. To address this issue, we examined the stability of avian malarial parasite lineages across multiple time-scales within two insular host communities. Parasite communities in Puerto Rico and St Lucia included 20 and 14 genetically distinct parasite lineages, respectively. Lineage composition of the parasite community in Puerto Rico did not vary seasonally or over a 1 year interval. However, over intervals approaching a decade, the avian communities of both islands experienced an apparent loss or gain of one malarial parasite lineage, indicating the potential for relatively frequent lineage turnover. Patterns of temporal variation of parasite lineages in this study suggest periodic colonization and extinction events driven by a combination of host-specific immune responses, competition between lineages and drift. However, the occasional and ecologically dynamic lineage turnover exhibited by insular avian parasite communities is not as rapid as antigenic allele turnover within populations of human malaria. PMID- 15129960 TI - An epidemiological model for West Nile virus: invasion analysis and control applications. AB - Infectious diseases present ecological and public health challenges that can be addressed with mathematical models. Certain pathogens, however, including the emerging West Nile virus (WN) in North America, exhibit a complex seasonal ecology that is not readily analysed with standard epidemiological methods. We develop a single-season susceptible-infectious-removed (SIR) model of WN cross infection between birds and mosquitoes, incorporating specific features unique to WN ecology. We obtain the disease reproduction number, R0, and show that mosquito control decreases, but bird control increases, the chance of an outbreak. We provide a simple new analytical and graphical method for determining, from standard public health indicators, necessary mosquito control levels. We extend this method to a seasonally variable mosquito population and outline a multi-year model framework. The model's numerical simulations predict disease levels that are consistent with independent data. PMID- 15129961 TI - Natural interspecific and intraspecific horizontal transfer of parthenogenesis inducing Wolbachia in Trichogramma wasps. AB - The intracellular bacterium Wolbachia is one of the most common symbionts in arthropods and, because of its manipulative effects on host reproduction, is assumed to be an important factor in several evolutionary processes. These bacteria are mainly vertically transmitted from mother to daughter through the egg cytoplasm, and horizontal transmission is generally assumed to be rare. Here, we show natural inter- and intraspecific horizontal transfer of parthenogenesis inducing Wolbachia between parasitoid wasps of the genus Trichogramma. Horizontal transfer was observed when infected and uninfected larvae shared the same host egg. This is the first report, to our knowledge, on interspecific horizontal transfer of Wolbachia between closely related sympatric species. Some originally uninfected immature wasps acquired Wolbachia while inside the host egg, but not all of these newly infected females exhibited the parthenogenesis phenotype. In general, intraspecific horizontal transfer was more successful than interspecific transfer. Wolbachia underwent vertical transmission in the new species but the infection tended to be lost within several generations. Our results have important implications for understanding the evolution of Wolbachia-host associations. PMID- 15129962 TI - Notch sensitivity of mammalian mineralized tissues in impact. AB - The toughness of bone is an important feature in preventing it from fracturing. We consider the notch sensitivity in impact, and the associations between brittleness, notch sensitivity and post-yield energy absorption of mammalian mineralized tissues. Specimens of bone-like tissues covering a wide range of mineralization were broken, either notched or un-notched, in impact. The greater the mineral content, the greater was the notch sensitivity. Also, the more brittle tissues dissipated the least post-yield energy and were the most notch sensitive. It is suggested that since antler bone, the least mineralized of all known mammalian mineralized tissues, seems to be notch insensitive in impact, no adaptive purpose would be served by having mineralized tissues of a lower mineralization than antler. This may explain the lower cut-off in mineralization seen in mammals. PMID- 15129963 TI - The limits of elaboration: curved allometries reveal the constraints on mandible size in stag beetles. AB - Many studies have demonstrated the adaptive advantage of elaborate secondary sexual traits, but few if any have shown compelling evidence for the limits to the elaboration of these traits that must exist. We describe such evidence in the exaggerated mandibles of stag beetles. In 1932, Huxley showed that the slope of the allometric relationship between mandible length and body size in some stag beetles declines in the largest males. We show that this curvature is most pronounced in species with relatively long mandibles, consistent with the hypothesis that the decrease in slope is caused by the increasing costs of large mandibles, which ultimately limit their size. Increasing depletion of resources in the prepupa and pupa by the rapidly growing mandibles is the most likely way in which these costs are manifested. The curved allometries have two components: intraspecific mandible allometry is steepest among small males of the species with the longest mandibles, but shallowest among the largest males of those same species. These patterns suggest that selection continues to favour positive allometry in species that invest relatively more in weaponry despite the limits to mandible exaggeration being reached in the largest males. PMID- 15129964 TI - Ancient lakes as evolutionary reservoirs: evidence from the thalassoid gastropods of Lake Tanganyika. AB - Ancient lakes are often collectively viewed as evolutionary hot spots of diversification. East Africa's Lake Tanganyika has long been the subject of scientific interest owing to dramatic levels of endemism in species as diverse as cichlid fishes, paludomid gastropods, decapod and ostracod crustaceans and poriferans. It is the largest and deepest of the African rift lakes, and its endemic fauna has been presented with a stable inland environment for over 10 Myr, offering unique opportunities for within-lake diversification. Although astonishing diversification has been documented in the endemic cichlid fauna of the lake, similar patterns of rapid diversification have long been assumed for other groups. In contrast to this hypothesis of rapid speciation, we show here that there has been no acceleration in the rate of speciation in the thalassoid gastropods of the lake following lake colonization. While limited within-lake speciation has occurred, the dramatic conchological diversity of gastropods presently found within the lake has evolved from at least four major lineages that pre-date its formation by as much as 40 Myr. At the same time, a widespread group of African gastropods appears to have evolved from taxa presently found in the lake. While Lake Tanganyika has been a cradle of speciation for cichlid fishes, it has also been an important evolutionary reservoir of gastropod lineages that have been extirpated outside the basin. PMID- 15129965 TI - Phylogenetic position of the Pentastomida and (pan)crustacean relationships. AB - Pentastomids are a small group of vermiform animals with unique morphology and parasitic lifestyle. They are generally recognized as being related to the Arthropoda; however, the nature of this relationship is controversial. We have determined the complete sequence of the mitochondrial DNA (mtDNA) of the pentastomid Armillifer armillatus and complete or nearly complete mtDNA sequences from representatives of four previously unsampled groups of Crustacea: Remipedia (Speleonectes tulumensis), Cephalocarida (Hutchinsoniella macracantha), Cirripedia (Pollicipes polymerus) and Branchiura (Argulus americanus). Analyses of the mtDNA gene arrangements and sequences determined in this study indicate unambiguously that pentastomids are a group of modified crustaceans probably related to branchiurans. In addition, gene arrangement comparisons strongly support an unforeseen assemblage of pentastomids with maxillopod and cephalocarid crustaceans, to the exclusion of remipedes, branchiopods, malacostracans and hexapods. PMID- 15129966 TI - Complex biogeographic history of a Holarctic passerine. AB - Our analysis of the ND2 sequences revealed six clades within winter wrens (Troglodytes troglodytes). These clades corresponded to six geographical regions: western Nearctic, eastern Nearctic, eastern Asia, Nepal, Caucasus and Europe, and differed by 3-8.8% of sequence divergence. Differences among regions explained 96% of the sequence variation in winter wren. Differences among individuals within localities explained 3% of the sequence variation, and differences among localities within regions explained 1%. Grouping sequences into subspecies instead of localities did not change these proportions. Proliferation of the six clades coincided with Early and Middle Pleistocene glaciations. The distribution of winter wren clades can be explained by a series of five consecutive vicariant events. Western Nearctic wrens diverged from the Holarctic ancestor 1.6 Myr before the present time (MYBP). Eastern Nearctic and Palaearctic wrens diverged 1 MYBP. Eastern and western Palaearctic birds diverged 0.83 MYBP. Nepalese and east Asian wrens diverged 0.67 MYBP, and Caucasian birds diverged from European wrens 0.54 MYBP. The winter wren has a much greater degree of inter- and intracontinental differentiation than the three other Holarctic birds studied to date--dunlin (Calidris alpina), common raven (Corvus corax) and three-toed woodpecker (Picoides trydactylus)--and represents an example of cryptic speciation that has been overlooked. PMID- 15129967 TI - Biopharmaceutic and pharmacokinetic aspects of variable bioavailability of rifampicin. AB - Even today the treatment outcome of tuberculosis is questionable due to variable bioavailability of rifampicin, which was discovered four decades back. In this manuscript, results of bioequivalence trials reported are presented in the form of a figure that provides a comprehensive look at the rifampicin bioavailability literature, provides understanding of the problem and clears 'myths and assumptions' regarding rifampicin bioavailability from fixed-dose combination (FDC) formulations. It was found that FDCs of good as well as bad quality rifampicin containing formulations with reduced or increased relative bioavailability are available. In addition, 'rifampicin alone' formulations also show variability in bioavailability. In the context of anomalous bioavailability of rifampicin, reasons postulated in literature are summarized. Approaches needed to solve the issue of rifampicin bioavailability are discussed on the basis of LADMER and BCS. PMID- 15129968 TI - The interaction of the penetration enhancer DDAIP with a phospholipid model membrane. AB - Differential scanning calorimetry (DSC) was used to investigate the mechanism of action of a proprietary skin penetration enhancer, dodecyl-2-(N,N dimethylamino)propionate (DDAIP) in dipalmitoylphosphatidylcholine (DPPC) liposomes. Furthermore, the effect of enhancer concentration on lipid thermotropic transitions was investigated. With increasing concentrations of DDAIP (from 5 to 50 mol%), the main transition peak shifted to lower temperatures and became more broad. The pretransition peak also shifted to lower temperatures with increasing concentrations of DDAIP and disappeared completely above an enhancer concentration of 20 mol%. Main transition and pretransition enthalpies of reaction decreased with increasing DDAIP concentration, indicating that enhancer treatment destabilized both rippled gel and liquid crystal phases within the bilayer. At and above a DDAIP concentration of 33.3 mol%, an additional transition was evident, indicating the presence of two phases of enhancer-lipid complex. Results suggest that DDAIP enhances drug transport by interacting with the polar region of the phospholipid bilayer and also by increasing the motional freedom of lipid hydrocarbon chains. PMID- 15129969 TI - Characterization of caprylocaproyl macrogolglycerides based microemulsion drug delivery vehicles for an amphiphilic drug. AB - Microemulsion systems composed of water, isopropyl myristate, PEG-8 caprylic/capric glycerides (Labrasol), and polyglyceryl-6 dioleate (Plurol Oleique), were investigated as potential drug delivery vehicles for an amphiphilic model drug (diclofenac diethylamine). Pseudo-ternary phase diagram of the investigated system, at constant surfactant/cosurfactant mass ratio (Km 4:1) was constructed at room temperature by titration, and the oil-to surfactant/cosurfactant mass ratios (O/SC) that exhibit the maximum in the solubilization of water were found. This allowed the investigation of the continuous structural inversion from water-in-oil to oil-in-water microemulsions on dilution with water phase. Furthermore, electrical conductivity (sigma) of the system at Km 1:4, and O/SC 0.250 was studied, and the percolation phenomenon was observed. Conductivity and apparent viscosity (eta') measurement results well described colloidal microstructure of the selected formulations, including gradual changes during their formation. Moreover, sigma, eta', and pH values of six selected microemulsion vehicles which differ in water phase volume fraction (phi(w)) at the selected Km and O/SC values, were measured. In order to investigate the influence of the amphiphilic drug on the vehicle microstructures, each system was formulated with 1.16% (w/w) diclofenac diethylamine. Electrical conductivity, and eta' of the investigated systems were strongly affected by drug incorporation. The obtained results suggest that diclofenac diethylamine interacts with the specific microstructure of the investigated vehicles, and that the different drug release kinetics from these microemulsions may be expected. The investigated microemulsions should be very interesting as new drug carrier systems for dermal application of diclofenac diethylamine. PMID- 15129970 TI - Structure-activity relationship of coordinated catecholamine in the [Ru(III)(NH3)4(catecholamine)]+ complex. AB - The redox chemistry and pharmacological studies of the novel blue ruthenium(III) catecholamine complexes were investigated in aqueous medium and compared to the free catecholamines. The [Ru(III)(NH3)4(catecholamine)]+ can be oxidized or reduced reversibly in one electron redox couples in aqueous solution. This is in contrast to the free catecholamines, which has a complicated electrochemical behavior due to coupled protonation process. The introduction of the ruthenium group reduces the intrinsic efficacy of the studied catecholamines. The [Ru(III)(NH3)4(catecholamine)]+ complex aqueous medium is more stable than the free catecholamines ligand in the same conditions. PMID- 15129971 TI - The characterization and biodistribution of cefoxitin-loaded liposomes. AB - To conquer the clinical restriction of relative short half-life and poor tissue retaining activities, liposomes containing cefoxitin were prepared using three methods in this study. The physicochemical properties including cefoxitin encapsulation percentage, vesicle size, stability, as well as the in vivo biodistribution were studied. The highest entrapment percentage was observed by using reverse phase evaporation method, and the molar ratio of cefoxitin to phospholipids was 1:3, DMPC to cholesterol was 2:1, respectively. From the result of stability, the freeze-drying powder and then stored in the frozen condition of cefoxitin-loaded liposome was an ideal storage state. Accordingly, the formulation by reverse-phase evaporation method was selected to investigate the biodistribution of cefoxitin-loaded liposome and compared to free cefoxitin in rats. It was observed that the cefoxitin levels and the duration retained in the liver, spleen, and pancreas of liposome-injected animals were higher and longer than that of free cefoxitin-injected animals. The drug concentrations of bile after post-injection of liposomal cefoxitin at 0.5, 1 and 2 h were all approximately 2.7 times higher than that of free cefoxitin injection group. PMID- 15129972 TI - Improvement of stability and absorbability of dry insulin powder for inhalation by powder-combination technique. AB - The effect of pulmonary absorption enhancers on the stability of active ingredients is an important factor for successful inhalation therapy as well as the effect on pharmacological activity and safety. We examined the effect of pulmonary absorption enhancers on the stability of insulin in dry powders prepared by a spray-drying technique. Although the hypoglycemic effect was greatly improved when a dry insulin powder containing citric acid (MIC SD) was administered, insulin in the MIC SD was unstable compared with the other powders examined. Bacitracin and Span 85, which are potent pulmonary absorption enhancers of insulin formulated in solutions, showed no deteriorative effect on the stability of dry insulin powder. However, they did not improve the hypoglycemic effect of insulin in dry powders. We modified the insulin dosage form with citric acid to improve the insulin stability at room temperature without loss of hypoglycemic activity. MIC Mix was formulated as a combination of insulin powder (MI') and citric acid powder (MC). MIC Mix showed hypoglycemic activity comparable to MIC SD while the insulin stability was much better than that of MIC SD at a 60 degrees C/dry condition. However, moisture lowered the insulin stability and changed the particle morphology of MIC Mix with time at a 60 degrees C/75% relative humidity condition, suggesting that a package preventing moisture absorption was necessary for the MIC Mix powder. PMID- 15129973 TI - Melted glyceryl palmitostearate (GPS) pellets for protein delivery. AB - Lysozyme was incorporated into glyceryl palmitostearate (GPS) pellets by compression and melting at loadings of 2, 5 and 10% (w/w). Released lysozyme from both compressed and melted pellets showed good retention of enzymatic activity (>80% active). The percentage lysozyme recovered during in vitro release experiments, over 120 h, was significantly lower from the melted pellets (<15%) compared with compressed pellets (71-85%). Scanning electron microscopy suggested this difference in release was due to differences in porosity of the compressed and melted pellets. Inclusion of hydrophilic components, PEG 4000 and Gelucire 50/13, in the melted matrices increased the percentage of lysozyme released in vitro. Lysozyme released from GPS/PEG 4000 matrices showed good retention of enzymatic activity (>88% active) while that from GPS/Gelucire 50/13 showed reduced activity (68 and 51% active). PEG 4000 was not completely miscible with GPS at the concentrations studied and heterogenous systems resulted. At a loading of 20-35% (w/w) PEG 4000 in GPS greater than 80% of the incorporated lysozyme was released, indicating the likely achievement of interconnecting hydrophilic channels throughout the GPS matrix. In conclusion, melted GPS demonstrated potential as a matrix for the controlled release of proteins and release rates could be modified by inclusion of hydrophilic components. PMID- 15129974 TI - A kinetic study on the degradation of erythromycin A in aqueous solution. AB - The pH is a critical factor determining the rate of the degradation of erythromycin A in aqueous solutions. However, the kinetics of the acid- and base catalyzed degradation is still uncertain. This study used a sensitive coulometric detection method to determine concentrations of erythromycin A and its degradation products. To determine the buffer-independent rate constants, sodium acetate (0.05-0.2 M) and Tris-HCl (0.1-0.5 M) were used in a pH range of 3.5-5.5 and 7.0-9.0, respectively. In acidic conditions, anhydroerythromycin A appeared to be produced directly through an internal dehydration of erythromycin A-6,9 hemiketal which simultaneously established an equilibrium with erythromycin A enol ether on the other hand. In weakly alkaline conditions, hydroxide ion appeared to catalyze the hydrolysis of the lactonyl ester bond of erythromycin A 6,9-hemiketal by the pseudo-first-order kinetics, and the C13 --> C11 translactonization and internal dehydration reactions subsequently occurred to form pseudoerythromycin A enol ether. We suggest here a predictive model for reasonable interpretation of the kinetics of erythromycin A degradation in aqueous solutions, in which the observed rate constant was expressed by the sum of the partial reaction rate constants for the acid- and base-catalyzed degradation of erythromycin A-6,9-hemiketal as a function of pH in a range of 3.0 10.0. PMID- 15129975 TI - Bioavailability of ibuprofen from hot-melt extruded mini-matrices. AB - The bioavailability of ibuprofen from hot-melt extruded mini-matrices based on ethyl cellulose and a hydrophilic excipient was tested. During the in vivo evaluation an oral dose of 300 mg ibuprofen was administered to healthy volunteers (n = 9) in a randomized cross-over study and compared with a commercially available sustained release product (Ibu-slow). The plasma samples were analysed by a validated HPLC-UV method. One mini-matrix formulation (F-1) consisted of 30% ibuprofen, 35% ethyl cellulose and 35% hydroxypropyl methylcellulose (Metolose 60 SH 50), while the second formulation (F-2) contained 60% ibuprofen, 20% ethyl cellulose and 20% xanthan gum. These mini-matrices were administered in hard gelatine capsules. Both formulations behaved in vivo as sustained release formulations with an HVD(t50% Cmax) value (time span during which the plasma concentration is at least 50% of the Cmax value) of 7.6 and 12.0 h for formulations F-1 and F-2, respectively, whereas a value of 5.2 h was obtained for Ibu-slow. Although a significantly higher Cmax and AUC(0-24 h) was seen for the reference product, the relative bioavailability of both experimental formulations was about 80%. PMID- 15129976 TI - Optimum formulation for sustained-release insulin. AB - Our aim was to prepare an optimum formulation for a sustained-release preparation of insulin using biodegradable polymer composed of co-poly(D,L-lactic/glycolic) acids (PLGA)(L/G ratio: 50/50). Various kinds of PLGA microcapsules containing 3% insulin were administered subcutaneously (250 U/kg) as a single dose to rats with streptozotocin-induced diabetes, and plasma insulin levels were monitored. The following results were obtained. (1) Glycerin and water were suitable additives to prepare a reproducible injectable formulation. (2) Addition of zinc compounds was essential to diminish rapid insulin release and six-fold molar excess of ZnO to insulin was desirable. (3) The size of insulin particles showed the following order: human insulin > lyophilized human insulin > Zn-free human insulin. Zn-free insulin was similar to lyophilized insulin with respect to control of rapid release, so a smaller particle size was essential. (4) The size of the microcapsules also affected the release of insulin. With larger microcapsules (approximately 30 microm), there was gradual release and a significant second phase of insulin release, while smaller microcapsules did not allow sustained release. Some variation in microcapsule size contributed to more constant and sustained release. (5) Based on the insulin release profile in vivo, a suitable molecular weight for PLGA was around 6000. The biological activity of insulin extracted from the formulation was similar to that of normal insulin. These experiments allowed us to prepare a desirable sustained-release insulin formulation. PMID- 15129977 TI - Ascorbyl palmitate vesicles (Aspasomes): formation, characterization and applications. AB - Vesicles with biological activity or with a targeting function in addition to carrier properties will have an added advantage. Vesicles prepared with amphiphiles having antioxidant property may have potential applications towards disorders implicated with reactive oxygen species. Ascorbyl palmitate (ASP) was explored as bilayer vesicle forming material. It formed vesicles (Aspasomes) in combination with cholesterol and a negatively charged lipid (dicetyl phosphate). Aspasomes were prepared by film hydration method followed by sonication in which aqueous azidothymidine (AZT) solution was encapsulated in aqueous regions of bilayer. Aspasomes were obtained with all compositions containing 18-72 mol% cholesterol. Differential scanning calorimetric data of aspasome dispersion and anhydrous mixtures of ascorbyl palmitate, cholesterol and dicetyl phosphate confirm the formation of bilayered vesicles with ascorbyl palmitate. Cholesterol content in aspasome did not exhibit any relation with vesicle size, zeta potential or percent entrapment. A substantial change in release rate of azidothymidine from aspasome was noticed on varying the proportion of cholesterol. Release rate and cholesterol content in Aspasomes did not exhibit any relation. A preparation with 45 mol% of cholesterol showed maximum retardation in release rate, than other compositions. The change in capture volume with time (latency) was studied for 8 h and with such a short duration study it was difficult to predict long term stability of these vesicles. But release experiments do indicate stability up to 18 h. Percent reducing activity of aspasome was estimated by measuring the absorbance of alpha,alpha-diphenyl beta-picrylhydrazyl (DPPH) at 517 nm after addition of test antioxidant samples. These studies revealed that the antioxidant potency of ascorbyl moiety is retained even after converting ascorbyl palmitate into vesicles (Aspasomes). The antioxidant potency of Aspasomes was assessed by measuring the protection offered by this preparation against quinolinic acid induced lipoperoxidation of whole human blood in vitro, where in the lipoperoxidation was monitored by measuring thiobarbituric acid reactive substances (TBARS) levels. Aspasome rendered much better antioxidant activity than ascorbic acid. Transdermal permeation of aspasomal AZT, ASP-AZT aqueous dispersion and AZT-solution across excised rat skin was investigated in vitro using Franz diffusion cell. Permeation of aspasomal AZT was much higher than the other two preparations. However, ASP-AZT aqueous dispersion has also enhanced permeation of AZT significantly over the AZT solution, indicating skin permeation enhancing property of ascorbyl palmitate. PMID- 15129978 TI - Co-encapsulation of isoniazid and rifampicin in liposomes and characterization of liposomes by derivative spectroscopy. AB - Taking into consideration the benefits of the combined therapy of isoniazid (INH) and rifampicin (RIF), this study focused on co-encapsulation of INH and RIF in the same liposome formulation. INH was incorporated in the aqueous phase and RIF in the lipid layer. Liposomes containing either INH or RIF were also prepared. All liposome formulations were compared for their loading capacity, encapsulation percentage and release properties. Drug amounts in the liposomes were estimated using peak-to-peak first-order derivative UV spectroscopy. Among the liposome formulations DPPC:chol liposomes showed the highest loading capacity (106.70 +/- 0.12 for INH and 18.17 +/- 0.06 (x 10(-3)) for RIF) and encapsulation percentage (73.84 +/- 0.78 for INH and 81.53 +/- 2.06 for RIF) compared to EPC:chol liposomes (loading capacity 93.36 +/- 0.58 for INH and 17.87 +/- 0.11 (x 10(-3)) for RIF; encapsulation percentage 64.61 +/- 0.51 for INH and 74.45 +/- 0.48 for RIF). Co-encapsulation of INH and RIF increased their individual encapsulation percentage and extended drug release compared to the formulations containing drug alone (Table 2). Results of this study support the conclusion that lipid and water soluble drugs can be successfully co-encapsulated in the same liposome formulation and also show that derivative UV spectroscopy is a sensitive method for direct and accurate quantification of these co-encapsulated drugs. PMID- 15129979 TI - Influence of the co-encapsulation of different excipients on the properties of polyester microparticle-based vaccine against brucellosis. AB - This work evaluates the influence of different pharmaceutical auxiliaries (Pluronic F68, polyvinylpyrrolidone [PVP] or Tween 20), when mixed with an antigenic extract from Brucella ovis (hot saline; HS), on the characteristics of the resulting poly(epsilon-caprolactone) (PEC) and poly(lactide-co-glycolide) (PLGA) microparticles. In all cases, PEC microparticles were smaller than PLGA ones. Concerning the HS loading, PLGA microparticles were highly dependent on the type of the excipient used, whereas all the PEC formulations displayed similar encapsulation efficiencies. For both types of microparticles, the presence of PVP induced a burst release effect. On the contrary, the use of Tween 20 or Pluronic F68 dramatically modified this profile. For PLGA-Tween 20 and PEC-Pluronic F68 microparticles, the HS was released in a pulsatil way during the first 7 days followed by a continuous release for at least 3 weeks. The antigenicity of the HS components was kept in all cases. Phagocytosis by murine monocytes showed a clear difference based just on the hydrophobicity of the polymer, being PEC microparticles better engulfed. Cell activation quantified by the release of H2O2 did not showed major differences between batches, however, microparticles of PEC and Pluronic F68 induced the highest nitric oxide production. Together, these results confirm the advantageous qualities of the "HS-PEC-Pluronic F68 microparticles" as favorable candidate for vaccine purposes against brucellosis. PMID- 15129980 TI - Retention and distribution of two 99mTc-DTPA labelled vaginal dosage forms. AB - To objectively evaluate the performance of new vaginal dosage forms, it is important to determine their time of residence and their distribution. This paper describes the in vivo characteristics of a reference and test product in this situation. METHOD: A randomised cross-over study was performed in the same phase of the menstrual cycle in eight pre-menopausal women. The retention and distribution of a commercially available vaginal clotrimazole cream and a test gel product, each "labelled" with 99mTc-DTPA was assessed by gamma scintigraphy for 24 h after administration of the products. Mass balance analysis was attempted by collecting and counting sanitary napkins worn for the study time. RESULTS: Within individuals there was little variation in the clearance of the formulations, but wide variation between individuals with a range between 81 and 1% of the administered doses retained by 24 h. The losses appeared to occur mainly at times of urination with 12 +/- 8% (cream) and 20 +/- 23% (gel) collected on the sanitary napkins, but 46 +/- 34% (cream) and 38 +/- 22% gel activity not accounted for by 24 h. The intravaginal distribution of activity was similar for each product. CONCLUSIONS: Radioactive tracer methods are useful in assessing and comparing vaginal dosage forms. PMID- 15129981 TI - Rapid dissolving high potency danazol powders produced by spray freezing into liquid process. AB - The objective of this study was to investigate the use of organic solvents in the spray freezing into liquid (SFL) particle engineering process to make rapid dissolving high potency danazol powders and to examine their particle size, surface area and dissolution rate. The maximum drug potency produced was 91% for SFL micronized danazol/PVP K-15. XRD indicated that danazol in the high potency SFL powders was amorphous. SEM micrographs revealed that the SFL danazol/PVP K-15 nanostructured aggregates had a porous morphology and were composed of many smooth primary nanoparticles with a diameter of about 100 nm. Surface areas of SFL danazol/PVP K-15 high potency powders were in the range of 28-115 m2/g. The SFL powders exhibited significantly enhanced dissolution rates. The rate of dissolution of micronized bulk danazol was slow; only 30% of the danazol was dissolved in 2 min. However, 95% of danazol was dissolved in only 2 min for the SFL high potency powders. The SFL process offers a highly effective approach to produce high potency danazol nanoparticles contained in larger structured aggregates with rapid dissolution rates, and is especially applicable to delivery systems containing poorly water soluble drugs. PMID- 15129982 TI - Effect of cyclodextrins on the complexation and transdermal delivery of bupranolol through rat skin. AB - Bupranolol (BPL) is a potent beta-blocking agent, the extensive first-pass metabolism (>90%) and rapid elimination half-life (1.5-2.0 h) of this drug make it well suited to be developed as a transdermal delivery system (TDS). Hydroxypropyl betaCD (HPbetaCD) and partially methylated betaCD (PMbetaCD) were used as penetration enhancers for BPL. The formation of inclusion complex of BPL with these cyclodextrins (CDs) was characterized in solution and solid states by phase solubility, X-ray diffractometry and differential scanning calorimetry (DSC) analyses. The effect of CDs on the permeation enhancement of BPL through rat skin was studied using side-by-side diffusion cells and pH 7.4 phosphate buffered saline (PBS). CDs were employed at different concentrations with 0.4% (w/v) BPL as well as with excess quantity of BPL (1.0%, w/v) that CDs could not complex all the BPL and the drug was in the form of an aqueous suspension. The permeation of BPL from its aqueous suspension (0.4%, w/v) significantly increased when CDs were used at low concentrations (up to 2 and 5%, w/v concentration for HPbetaCD and PMbetaCD, respectively) (P < 0.01). At higher CD concentrations, the permeation of BPL decreased; and both CDs at 10% (w/w), showed similar flux values to that of control (no enhancer, P > 0.05). The permeation of BPL from its 1.0% (w/v) aqueous suspension increased with increase in concentration of CD up to 10% (w/v) for HPbetaCD and PMbetaCD. At 10% (w/v) concentration of HPbetaCD and PMbetaCD, the flux of BPL from its 1.0% aqueous suspension increased 3.8- and 4.6-fold (P < 0.01 and P < 0.001, respectively). The permeation data of skin pretreatment with CDs indicate that HPbetaCD had no effect on the skin, whereas PMbetaCD significantly reduced the skin barrier for BPL, as shown by 1.7-fold increase in the flux by PMbetaCD pretreatment (P < 0.001). Overall, both HPbetaCD and PMbetaCD were found to be suitable for improving the solubility and penetration enhancement of BPL. PMID- 15129983 TI - Amphiphilic dextran-graft-poly(epsilon-caprolactone) films for the controlled release of paclitaxel. AB - Amphiphilic graft copolymers with dextrans as the main chains and poly(epsilon caprolactone) as the side chains were synthesized by solution polymerization in dimethyl sulfoxide using stannous 2-ethylhexanoate as a catalyst. The copolymers were characterized with nuclear magnetic resonance spectroscopy and gel permeation chromatography. Paclitaxel loaded copolymer films were prepared using a solution cast method. In vitro release rates were influenced by both the initial paclitaxel loading and the morphology of the films. The films with paclitaxel loadings up to 10% (w/w) showed a single-phase morphology when dry. However, paclitaxel crystallization occurred in the films with high loadings (5 and 10%) upon incubation in phosphate buffered saline at 37 degrees C. The crystals had a spherulitic structure and were found to be paclitaxel dihydrate based on X-ray diffraction analysis. The formation of paclitaxel dihydrate in the films produced a marked effect on the in vitro release rates. PMID- 15129984 TI - Assessing aggregation of peptide conjugate of doxorubicin using quasi-elastic light scattering and 600 MHz NMR. AB - The use of doxorubicin in treating prostate cancer is limited by its systemic toxicities especially cardiotoxicity and immunosuppression. Prodrugs that reduce the systemic exposure of doxorubicin are believed to provide a safety advantage. A prodrug of doxorubicin which contains a peptide sequence that can be recognized by prostate-specific antigen (PSA) and cleaved in the prostate was formulated for clinical use. The i.v. formulation and manufacture of this peptide conjugate posed several challenges. The main issue of the i.v. formulation were chemical and physical stability. The physical stability challenges posed during formulation and manufacture of his peptide conjugate is described herein. A heptapeptide conjugate of doxorubicin was found to aggregate in solution forming large ill defined aggregates (60-1300 nm). In contrast to doxorubicin, the average hydrodynamic diameter measured for this compound by dynamic laser light scattering technique is very large. Increasing concentration of the drug and lowering pH promoted aggregation. We rationalize the difference in the effective hydrodynamic diameter due to hydrogen bonding of the peptide which allows for the formation of large particle sizes relative to doxorubicin. We have also used 600 MHz 1H NMR to assess the aggregation of this compound. PMID- 15129985 TI - Controlled release of a model protein lysozyme from phase sensitive smart polymer systems. AB - The purpose of this study was to investigate the suitability of phase sensitive smart polymer-based protein formulations in order to deliver a model protein, lysozyme, in a conformationally stable and biologically active form at a controlled rate over extended period of time. Four different formulations, using D,L-poly(lactide) (D,L-PLA) and a solvent mixture of different ratios of benzyl benzoate (BB) and benzyl alcohol (BA), were prepared. Conformational stability and biological activity of lysozyme were studied by differential scanning calorimeter and enzyme activity assay, respectively. We found a significant (P < 0.05) increase in burst and rate of release of incorporated lysozyme from formulations containing greater proportion of BA. In order to increase the conformational stability and biological activity of lysozyme, we incorporated mannitol as stabilizer into formulations. Mannitol increased the conformational and biological activity of lysozyme in comparison to the control formulation prepared without mannitol. In conclusion, phase sensitive smart polymer-based delivery systems were able to deliver a model protein, lysozyme, in a conformationally stable and biologically active form at a controlled rate over extended period of time. PMID- 15129986 TI - Evaluation of xanthan and highly substituted galactomannan from M. scabrella as a sustained release matrix. AB - A highly substituted galactomannan (G) from Mimosa scabrella Bentham (Man:Gal 1.1:1), isolated from the seeds of a Brazilian leguminous tree and xanthan (X), an exopolysaccharide secreted by Xanthomonas campestris (Keltrol), were evaluated as a hydrophilic matrix system (XG) for controlled release (CR) of diclofenac sodium (DS) in tablets and capsules. The performance of XG (2:1) matrices containing 50 mg (A) or 100 mg (B) of DS was compared with a commercial CR product of DS. The drug release studies were carried out using a dissolution apparatus (paddle method) with gradual increase of pH values, from pH 1.4, to pH 4.0 (after 1 h) and to pH 6.8 (after 2 h). The results suggested the potential of XG systems as release retarding materials, which released 78.6 and 35.1% of drug after 24 h for capsules (A) and tablets (A), respectively. Drug release decreased with the increase of amount of drug and it is dependent of dosage form. Analysis of release data indicate a rather zero-order drug release with the erosion mechanism playing a dominant role. PMID- 15129987 TI - Critical effect of freezing/freeze-drying on sustained release of FITC-dextran encapsulated within PLGA microspheres. AB - The cause of initial burst release of hydrophilic macromolecular drugs from biodegradable polymeric microspheres was identified. Poly(D,L-lactic-co-glycolic acid) microspheres encapsulating fluorescein isothiocyanate (FITC)-labled dextran was prepared by a double emulsion solvent evaporation method. The extent of initial burst release was examined by varying the formulation process conditions such as solvent evaporation, washing, freezing, and freeze-drying. Confocal microscopy was employed to analyze the underlying mechanism of burst release. The extent of burst release was gradually reduced after the repeated washing of embryonic microspheres before freeze-drying, indicating that FITC-dextran molecules entrapped within unhardened microspheres were slowly diffused out. However, freezing and subsequent drying processes of the embryonic microspheres resulted in much increased extent of burst release, suggesting that the initial burst release was primarily caused by the rapid diffusion of FITC-dextran through the microporous channels. Confocal microscopic analysis revealed that the freeze drying process generated water-escaping micro-channels, through which the encapsulated molecules were presumably dumped out. Vacuum-drying was a good alternative choice in reducing the initial burst, compared to freeze-drying. PMID- 15129988 TI - Measurement of density variations in tablets using X-ray computed tomography. AB - In this paper the application of X-ray computed tomography (CT) to measure the material density distribution in pharmaceutical tablets is discussed. X-ray CT is a non-destructive inspection technique which provides cross-sectional images in different planes through a component. The CT image values provide information on the local X-ray attenuation coefficients. For a particular material and X-ray energy, X-ray attenuation is approximately proportional to material density. Determination of quantitative density distributions requires consideration of non linear instrumental effects including scatter and "beam hardening". Density maps in tablets manufactured under controlled conditions are presented. The results are discussed with reference to the local properties of the material within the tablet and the tablet design features. PMID- 15129989 TI - In situ intestinal absorption studies on low molecular weight heparin in rats using labrasol as absorption enhancer. AB - Oral absorption of low molecular weight heparin (LMWH) is limited by its molecular size and negative charge. Development of its oral formulations would allow outpatient treatment with LMWH and decrease the hospital expenses. Studies were aimed at evaluating Labrasol for improving intestinal absorption of LMWH. Formulations containing LMWH and Labrasol were administered to duodenum, jejunum, and ileum of the fasted rats. The doses of LMWH and Labrasol were 200 IU/kg and 50 mg/kg, respectively. Reversibility of absorption enhancing effect of Labrasol was assessed by administering LMWH to jejunum after 0.5 and 1 h of administration of Labrasol. The effect of different doses of Labrasol on LMWH absorption was studied by administering Labrasol at 50, 100, and 200 mg/kg doses. Administration of LMWH formulation tojejunum resulted in the highest plasma anti-Xa activity (0.50+/-0.03 IU/ml) compared to duodenum (0.19+/-0.03 IU/ml), and ileum (0.29 +/ 0.06 IU/ml) and the anti-Xa levels were maintained above the therapeutic level for about 160 min. The absorption of LMWH was negligible when LMWH was administered at 0.5 and 1 h post-Labrasol administration. Increasing the dose of Labrasol has decreased the absorption of LMWH from jejunum. Labrasol increased the intestinal absorption of LMWH, and jejunum was found to be the best site of absorption. Intestinal membrane permeability changes induced by Labrasol were transient and reversible. Maintaining high drug concentration gradient across intestinal wall is important to obtain increased intestinal LMWH absorption. PMID- 15129990 TI - Oral sustained delivery of ambroxol from in situ-gelling pectin formulations. AB - Gels formed in situ following oral administration of dilute aqueous solutions of pectin (1.0 and 1.5%, w/v) to rats were evaluated as vehicles for the sustained release of the expectorant drug ambroxol hydrochloride. The solutions contained calcium ions in complexed form, which on release in the acidic environment of the stomach caused gelation of the pectin. In vitro studies demonstrated diffusion controlled release of ambroxol from the gels over a period of 6 h. A bioavailability of ambroxol of approximately 64% of that of a commercially available formulation could be achieved from gels containing an identical dose of ambroxol formed in situ in the stomachs of rats, with appreciably lower peak plasma levels and a sustained release of drug over a period of at least 6 h. The influence of added sorbitol (17%, w/v) on the rheological and drug release properties of the formulations has been examined. PMID- 15129991 TI - Use of the ninhydrin assay to measure the release of chitosan from oral solid dosage forms. AB - This study evaluated and optimised the ninhydrin assay as a tool for measuring the in vitro release and dissolution of chitosan from solid dosage forms. The precision and accuracy of the assay for the type of chitosan used in the study were examined by measuring the inter- and intra-sample variation and found to be within acceptable limits. The assay was applied practically to construct a pH/solubility profile for chitosan and subsequently to measure the release and dissolution of chitosan from dosage forms in the presence and absence of a model drug, sodium salicylate. Assay performance was found to be satisfactory over a wide range of physiologically relevant pH values. It is concluded that the ninhydrin assay is an essential aid in the design and testing of solid dosage forms with different chitosan-drug release profiles. PMID- 15129992 TI - Non-invasive assessment of the effect of formulation excipients on stratum corneum barrier function in vivo. AB - The objective of the present work was to investigate the effect of formulation excipients on human stratum corneum (SC) barrier function in vivo. Two formulations, an ointment and an oil-in-water cream, were applied to the skin of human volunteers under both occlusive and non-occlusive conditions. The effects of each treatment were then evaluated using three non-invasive biophysical techniques: transepidermal water loss (TEWL), impedance spectroscopy (IS) and attenuated-total-reflectance Fourier transform infrared (ATR-FTIR) spectroscopy. These measurements were combined with a simple tape-stripping protocol to allow information to be derived across the entire SC. IS and TEWL provided basic information on the effect of each formulation on skin barrier function, while ATR FTIR enabled (i) the tracking of formulation excipients and evaluation of their concentration profiles within the SC, and (ii) deduction of mechanistic detail with which to explain the TEWL and IS results. It was found that occlusion of the skin either in the presence or absence of the cream caused TEWL to be increased when the treatment was terminated at 6 h. Uptake of ointment into the SC, on the other hand, inhibited the post-application TEWL rate. In parallel, treatment with the ointment caused an increase in relative low-frequency skin impedance, consistent with the entry of additional lipophilic constituents into the SC. The latter was confirmed by ATR-FTIR spectroscopic measurements. Overall, the combined use of the three biophysical measurements allowed formulation effects on, and uptake into, the SC to be deduced and evaluated, and the approach may prove useful for the future selection and optimization of topical drug delivery vehicles. PMID- 15129993 TI - Influence of chitosan and its glutamate and hydrochloride salts on naproxen dissolution rate and permeation across Caco-2 cells. AB - Chitosan and its glutamate and hydrochloride salts were evaluated for their efficacy in improving the dissolution behaviour of naproxen (a poorly water soluble antiinflammatory drug) and its transport in vitro across Caco-2 cell monolayers. Drug-polymer physical mixtures and coground products, prepared at two different w/w ratios (30/70 and 10/90), were characterized by differential scanning calorimetry, X-ray powder diffractometry, scanning electron microscopy, and tested for dissolution properties. Coground systems were more effective than physical mixtures in improving drug dissolution and chitosan base, in spite of its lower water solubility, showed higher solubilizing power than its salts. According to the solid state analyses results, this effect was directly related to its stronger amorphizing power. Transport studies showed that only coground mixtures with chitosan glutamate salt allowed a significant drug apparent permeability improvement; however, they did not exhibit appreciable effects on the Caco-2 tight junctions (measured by the trans-epithelial electrical resistance variations), thus indicating that their enhancer effect was mainly due to an improved naproxen transport by transcellular passive diffusion rather than through the paracellular route. The direct compression properties and antiulcerogenic activity together with the demonstrated dissolution and permeation enhancer abilities toward naproxen make chitosan glutamate an optimal carrier for developing fast-action oral solid dosage forms of this drug. PMID- 15129994 TI - Vehicle-controlled effect of urea on normal and SLS-irritated skin. AB - It is known that, depending on the concentration, treatment with urea could improve skin barrier function, despite its penetration-enhancing properties. This controversial skin effect of urea has been explored systematically in this study in terms of the effect of vehicle on the performance of urea. In the first part, a series of four semi-solid emulsions with 5% (w/w) urea, varying in the type of emulsion, nature of emulsifier and polarity of oil ingredients, have been evaluated with regard to their skin hydrating and transepidermal water loss (TEWL)-modifying properties. Placebo samples were tested alongside the urea containing ones. Two best performing moisturisers from the above were chosen for the second part of the study, in which sodium lauryl sulphate (SLS)-irritated skin was treated with both placebo and urea-containing samples. In addition to TEWL and skin hydration level, the erythema index (EI) was measured before, during and after the treatment. The results have shown that barrier-improving and hydrating abilities of urea are bi-directional and dependent on both the type of vehicle used for its delivery and the state of skin. PMID- 15129995 TI - Characterization of curcumin-PVP solid dispersion obtained by spray drying. AB - Curcumin, a naturally occurring highly lipophilic molecule has wide range of pharmacological activities. However, its limited aqueous solubility and degradation at alkaline pH restricts its bioavailability. Solid dispersions of curcumin in different ratios with PVP were prepared by spray drying. Physical characterization by SEM, IR, DSC, and XRPD studies, in comparison with corresponding physical mixtures revealed the changes in solid state during the formation of dispersion and justified the formation of high-energy amorphous phase. Dissolution studies of curcumin and its physical mixtures in 0.1 N HCl showed negligible release even after 90 min. Whereas, solid dispersions showed complete dissolution within 30 min. This may aid in improving bioavailability and dose reduction of the drug. PMID- 15129996 TI - Microbiological assay for enrofloxacin injection. AB - A simple, sensitive and specific agar diffusion bioassay for the antibacterial enrofloxacin was developed. Using a strain of Staphylococcus aureus ATCC 6538P as the test organism, enrofloxacin at concentrations ranging from 3.2 to 12.8 microg ml(-1) could be measured in injection. A prospective validation of the method showed that method was linear (r = 0.99998), precise (R.S.D. = 0.27) and accurate (it measured the added quantities). The method shows results that confirm its precision, not differing significantly the other method described in the literature. We conclude that microbiological assay is satisfactory for quantitation of in vitro antibacterial activity of enrofloxacin. PMID- 15129997 TI - Calcium pterin as an antitumor agent. AB - A series of in vivo studies are reported that provide evidence for an immunologically mediated mechanism for the antitumor response from a calcium pterin (CaPterin) suspension. Strong antitumor efficacy was demonstrated in fully immunocompetent female C3H/HeN-MTV+ mice (retired breeders) presenting spontaneous mammary gland adenocarcinomas. Comparison of results obtained by testing CaPterin in either nude or SCID mice (severely compromised immunodeficient) implanted with MDA-MB-231 human cancer cells showed a significant antitumor response in the nudes and no response in the SCIDs. This comparison argues for B-cell immunological involvement in the mechanism of CaPterin antitumor activity since nude mice possess B-cell capability while SCID mice do not. This comparison also indicates that there is no measurable direct cancer cell toxicity from the CaPterin. Results showing no CaPterin antitumor efficacy against EMT6 tumor cells implanted in Balb/c mice also suggest an antitumor mechanism involving B-cells, since transforming growth factor beta (TGF beta), produced by EMT6 cells, is known to cause B-cell apoptosis. Taken together, these results, along with those of other researchers, indicate that CaPterin's antitumor mechanism involves antibody-dependent cellular cytotoxicity (ADCC) mediated, for example, by natural killer (NK) cells, interlukin-2, and CaPterin. PMID- 15129998 TI - In vitro skin penetration of dazmegrel delivered with a bioelastic matrix. AB - The penetration of dazmegrel, a selective thromboxane synthetase inhibitor, through excised human and greyhound skin was measured. A bioelastic matrix was used for topical delivery. Results demonstrated that dazmegrel readily penetrated the skin. Penetration through greyhound skin was significantly greater than penetration through human skin. Penetration through greyhound skin was not significantly different between 4, 24, and 48 h of exposure for the low and intermediate doses studied. PMID- 15129999 TI - Simultaneous permeation of tamoxifen and gamma linolenic acid across excised human skin. Further evidence of the permeation of solvated complexes. AB - Tamoxifen is the hormonal treatment of choice in women who have hormone-dependent breast cancer and its efficacy in those women considered to have a high risk of developing breast cancer, has also been established. Gamma linolenic acid (GLA) has been shown to decrease the invasion of breast cancer and recent studies have demonstrated that GLA can enhance the oestrogen receptor down-regulation induced by tamoxifen. However, tamoxifen is associated with serious side-effects due mainly to systemic delivery, and targeted delivery of both tamoxifen and GLA would be highly beneficial. This work was a preliminary study for the development of a transcutaneous system to simultaneously deliver both tamoxifen and GLA directly to the breast. Full thickness human skin was dosed with 500 microl saturated solution of tamoxifen in borage oil (25% GLA) and the simultaneous permeation of the two actives determined. There was rapid flux with minimal lag time, the cumulative permeation at 24 h was 764.3 +/- 94.2 microg cm(-2) for GLA and 5.44 +/- 0.67 microg cm(-2) for tamoxifen: the latter being comparable to the amount of tamoxifen associated with cancerous breast tissue from a 20 mg oral dose. The ratio of GLA/tamoxifen permeated at different timepoints was quite consistent, both in terms of mass (mean 138, S.D. 15.1) and mols (mean 184, S.D. 20.3). It was determined that 2.5 molecules of GLA were associated with each molecule of tamoxifen in the permeation process, equating to a solvation cage of three molecules of triacylglycerol. This study has demonstrated the feasibility of administering simultaneously tamoxifen and GLA using borage oil as vehicle, which warrants further investigation as a novel topical two-component system in relation to or prophylaxis of those perceived at high risk of developing breast cancer. The study also provides further evidence of the permeation of solvated complexes across skin, rather than discrete penetrant molecules. PMID- 15130000 TI - Diffraction effects on broadband radiation: formulation for computing total irradiance. AB - I present a formulation for treating diffraction effects on total irradiance in the case of a Planck source; earlier work generally depended on calculating diffraction effects on spectral irradiance followed by summation over spectral components. The formulation is derived and demonstrated for Fraunhofer diffraction by circular apertures, rectangular apertures and slits, and Fresnel diffraction by circular apertures. The prospects for treating other sources and optical systems are also discussed. PMID- 15130001 TI - Design and characterization of a photometer-colorimeter standard. AB - A photometer and tristimulus colorimeter has been developed at the National Institute of Standards and Technology (NIST) to realize a color scale. A novel construction was developed to implement the spectral-responsivity-based scale with small uncertainty. The new device can be used as a reference illuminance and luminance meter as well. Temperature-controlled filter combinations, with 5-8 layers in one package, are used to match the responsivity of a silicon tunnel trap detector to the CIE color-matching functions with small spectral mismatch values (f1'). Design considerations to extend the tunnel-trap detector with replaceable single and double apertures and changeable filter combinations are described. The design and fabrication of the filter packages and the dependence of the f1' values on the thickness of the filter layers are discussed. The colorimeter was characterized for angular, spatial, and spectral responsivity. An improved preamplifier can convert current to voltage in an 11-decade dynamic range with 0.01% uncertainty. PMID- 15130002 TI - Fabrication and alignment issues for segmented mirror telescopes. AB - There is a great demand for new telescopes that use larger primary mirrors to collect more light. Because of the difficulty in the fabrication of mirrors larger than 8 m as a single piece, they must be made with numerous smaller segments. The segments must fit together to create the effect of a single mirror, which presents unique challenges for fabrication and testing that are absent for monolithic optics. This is especially true for the case of a highly aspheric mirror required to make a short two-mirror telescope. We develop the relationship between optical performance of the telescope and errors in the manufacture and operation of the individual segments. PMID- 15130004 TI - Markov-chain Monte Carlo approach to the design of multilayer thin-film optical coatings. AB - We reconsider the problem of locating the globally optimal solution of a multilayer-optical-coating design problem, within some predetermined space of parameters, with the aim of obtaining a robust technique that requires a minimum of user intervention. The approach we adopt centers on exploring the space of the parameters of interest by using a Markov-chain Monte Carlo sampling algorithm. This technique enables one to locate the global optimum automatically with high confidence and without the need for a good starting design. It also allows the trivial inclusion of prior constraints on the variables and provides a natural means for investigating the robustness of the optimal solution. PMID- 15130003 TI - Skin cancer detection by spectroscopic oblique-incidence reflectometry: classification and physiological origins. AB - Data obtained from 102 skin lesions in vivo by spectroscopic oblique-incidence reflectometry were analyzed. The participating physicians initially divided the skin lesions into two visually distinguishable groups based on the lesions' melanocytic conditions. Group 1 consisted of the following two cancerous and benign subgroups: (1) basal cell carcinomas and squamous cell carcinomas and (2) benign actinic keratoses, seborrheic keratoses, and warts. Group 2 consisted of (1) dysplastic nevi and (2) benign common nevi. For each group, a bootstrap-based Bayes classifier was designed to separate the benign from the dysplastic or cancerous tissues. A genetic algorithm was then used to obtain the most effective combination of spatiospectral features for each classifier. The classifiers, tested with prospective blind studies, reached statistical accuracies of 100% and 95% for groups 1 and 2, respectively. Properties that related to cell-nuclear size, to the concentration of oxyhemoglobin, and to the concentration of deoxyhemoglobin as well as the derived concentration of total hemoglobin and oxygen saturation were defined to explain the origins of the classification outcomes. PMID- 15130005 TI - Design, deposition, and characterization of multilayer coatings for the ultraviolet and visible-light coronagraphic imager. AB - The Ultraviolet and Visible-Light Coronagraphic Imager is the baseline coronagraph for the European Space Agency's payload Solar Orbiter, a solar mission whose launch in 2011 is expected. To prove the feasibility of its innovative design, a sounding rocket version of the same instrument has been approved by NASA. One of the main technological challenges of the instrument is the achievement of multilayer optical coatings with suitable properties. We describe the design, fabrication, and characterization of such coatings. PMID- 15130006 TI - Mechanical characteristics of optical coatings prepared by various techniques: a comparative study. AB - Good performance of optical coatings depends on the appropriate combination of optical and mechanical properties. Therefore, successful applications require good understanding of the relationship between optical microstructural and mechanical characteristics and film stability. In addition, there is a lack of standard mechanical tests that allow one to compare film properties measured in different laboratories. We give an overview of the methodology of mechanical measurements suitable for optical coatings; this includes depth-sensing indentation, scratch resistance, friction, abrasion and wear testing, and stress and adhesion evaluation. We used the techniques mentioned above in the same laboratory to systematically compare the mechanical behavior of frequently used high- and low-index materials, namely, TiO2, Ta2O5, and SiO2, prepared by different complementary techniques. They include ion-beam-assisted deposition by electron-beam evaporation, magnetron sputtering, dual-ion-beam sputtering, plasma enhanced chemical-vapor deposition, and filtered cathodic arc deposition. The mechanical properties are correlated with the film microstructure that is inherently related to energetic conditions during film growth. PMID- 15130007 TI - Design of a mechanical-tunable filter spectrometer for noninvasive glucose measurement. AB - The development of an accurate and reliable noninvasive near-infrared (NIR) glucose sensor hinges on the success in addressing the sensitivity and the specificity problems associated with the weak glucose signals and the overlapping NIR spectra. Spectroscopic hardware parameters most relevant to noninvasive blood glucose measurement are discussed, which include the optical throughput, integration time, spectral range, and the spectral resolution. We propose a unique spectroscopic system using a continuously rotating interference filter, which produces a signal-to-noise ratio of the order of 10(5) and is estimated to be the minimum required for successful in vivo glucose sensing. Using a classical least-squares algorithm and a spectral range between 2180 and 2312 nm, we extracted clinically relevant glucose concentrations in multicomponent solutions containing bovine serum albumin, triacetin, lactate, and urea. PMID- 15130008 TI - Six-degrees-of-freedom alignment technique that provides diagnostic misalignment information. AB - We introduce a new six-degrees-of-freedom alignment technique that enables the precise alignment of two optical device planes. The method combines linear diffraction gratings and cylindrical Fresnel lenses that allow the diagnosis of misalignment in each degree of freedom independently. The technique was used to align two 20 mm x 20 mm fused-silica substrates separated by 17 mm. The worst case alignment precision was found to be better than +/- 5 microm laterally, +/- 20 microm longitudinally, +/- 0.036 degrees rotationally, and +/- 0.007degrees in tilt. PMID- 15130009 TI - Windowed Fourier transform for fringe pattern analysis. AB - Fringe patterns in optical metrology systems need to be demodulated to yield the desired parameters. Time-frequency analysis is a useful concept for fringe demodulation, and a windowed Fourier transform is chosen for the determination of phase and phase derivative. Two approaches are developed: the first is based on the concept of filtering the fringe patterns, and the second is based on the best match between the fringe pattern and computer-generated windowed exponential elements. I focus on the extraction of phase and phase derivatives from either phase-shifted fringe patterns or a single carrier fringe pattern. Principles as well as examples are given to show the effectiveness of the proposed methods. PMID- 15130010 TI - Space-multiplexed optical scanner. AB - A low-loss two-dimensional optical beam scanner that is capable of delivering large (e.g., > 10 degrees) angular scans along the elevation as well as the azimuthal direction is presented. The proposed scanner is based on a space switched parallel-serial architecture that employs a coarse-scanner module and a fine-scanner module that produce an ultrahigh scan space-fill factor, e.g., 900 x 900 distinguishable beams in a 10 degrees (elevation) x 10 degrees (azimuth) scan space. The experimentally demonstrated one-dimensional version of the proposed scanner has a supercontinuous scan, 100 distinguishable beam spots in a 2.29 degrees total scan range, and 1.5-dB optical insertion loss. PMID- 15130011 TI - Phase plate to extend the depth of field of incoherent hybrid imaging systems. AB - A hybrid imaging system combines a modified optical imaging system and a digital postprocessing step. We describe a spatial-domain method for designing a pupil phase plate to extend the depth of field of an incoherent hybrid imaging system with a rectangular aperture. We use this method to obtain a pupil phase plate to extend the depth of field, which we refer to as a logarithmic phase plate. Introducing a logarithmic phase plate at the exit pupil of a simulated diffraction-limited system and digitally processing the detector's output extend the depth of field by an order of magnitude more than the Hopkins defocus criterion. We also examine the effect of using a charge-coupled device optical detector, instead of an ideal optical detector, on the extension of the depth of field. Finally, we compare the performance of the logarithmic phase plate with that of a cubic phase plate in extending the depth of field of a hybrid imaging system with a rectangular aperture. PMID- 15130012 TI - Effective spherical aberration compensation by use of a nematic liquid-crystal device. AB - The next generation of optical data storage system beyond DVDs will use blue laser light and an objective lens with a high numerical aperture of 0.85 to increase storage capacity. Such high numerical aperture systems have an inherent higher sensitivity to aberrations. In particular, the spherical aberration caused by cover layer thickness tolerances and--more obvious--by dual-layer disks with a typical separation of approximately 20 microm between the two layers must be compensated. We propose a novel transmissive nematic liquid-crystal device, which is capable of compensating spherical aberration that occurs during the operation of optical pickup systems. PMID- 15130013 TI - 0.18-microm complementary metal-oxide semiconductor push-pull vertical-cavity surface-emitting laser driver operating at 2.5 Gb/s with symmetric rising and falling edges. AB - A vertical-cavity surface-emitting laser (VCSEL) driver design that utilizes a novel push-pull circuit topology is described. The VCSEL driver design can provide both a current pushing and a current pulling mechanism and therefore is capable of producing symmetric rise and fall times. The design was implemented in a 0.18-microm foundry n-well complementary metal-oxide semiconductor technology and operates at data rates up to 2.5 Gb/s with a power consumption of 45 mW at an average optical output power of 1 mW. PMID- 15130014 TI - Hybrid numerical method for solution of the radiative transfer equation in one, two, or three dimensions. AB - A hybrid method is presented by which Monte Carlo (MC) techniques are combined with an iterative relaxation algorithm to solve the radiative transfer equation in arbitrary one-, two-, or three-dimensional optical environments. The optical environments are first divided into contiguous subregions, or elements. MC techniques are employed to determine the optical response function of each type of element. The elements are combined, and relaxation techniques are used to determine simultaneously the radiance field on the boundary and throughout the interior of the modeled environment. One-dimensional results compare well with a standard radiative transfer model. The light field beneath and adjacent to a long barge is modeled in two dimensions and displayed. Ramifications for underwater video imaging are discussed. The hybrid model is currently capable of providing estimates of the underwater light field needed to expedite inspection of ship hulls and port facilities. PMID- 15130015 TI - Quadratic behavior of fiber Bragg grating temperature coefficients. AB - We describe the characterization of the temperature and strain responses of fiber Bragg grating sensors by use of an interferometric interrogation technique to provide an absolute measurement of the grating wavelength. The fiber Bragg grating temperature response was found to be nonlinear over the temperature range -70 degrees C to 80 degrees C. The nonlinearity was observed to be a quadratic function of temperature, arising from the linear dependence on temperature of the thermo-optic coefficient of silica glass over this range, and is in good agreement with a theoretical model. PMID- 15130016 TI - Hyperspectral imager, from ultraviolet to visible, with a KDP acousto-optic tunable filter. AB - Hyperspectral imaging in the ultraviolet to visible spectral region has applications in astronomy, biology, chemistry, medical sciences, etc. A novel electronically tunable, random-wavelength access, compact, no-moving-parts, vibration-insensitive, computer-controlled hyperspectral imager operating from 220 to 480 nm with a spectral resolution of 160 cm(-1), e.g., 2 nm at 350 nm, has been developed by use of a KDP acousto-optic tunable filter (AOTF) with an enhanced CCD camera and a pair of crossed calcite Glan-Taylor polarizing prisms. The linear and angular apertures of the AOTF are 1.5 x 1.5 cm2 and 1.2 degrees, respectively. Imager setup and spectral imaging results as well as analyses and discussion of various factors affecting image quality are presented. PMID- 15130017 TI - High power density beam from narrow diode-laser arrays in axial symmetry. AB - A multiplexing method based on narrow diode-laser arrays (DLAs) in an axially symmetric configuration is described. The use of submillimeter narrow DLAs improves beam quality considerably in the slow-axis direction compared with typical 1-cm-wide DLAs. The axially symmetric geometry is advantageous for efficient spatial, wavelength, and polarization multiplexing. With narrow DLAs and an axially symmetric geometry, a small circular focus with more than 10 kW/mm2 average power density is possible. Theoretical calculations and preliminary experimental results are presented. PMID- 15130018 TI - Noise assessment of a Fourier transform infrared spectroradiometer subject to the stability of a conventional laboratory blackbody source. AB - A method is described for the measurement of the noise-equivalent spectral radiance (NESR) of Fourier transform infrared (FTIR) spectroradiometers at all wave numbers of a selected range. The method requires minimal detailed knowledge of the sensor and no support equipment beyond a blackbody source. The NESRs of the FTIR spectroradiometer are determined at every wave-number increment in the 700-1300 cm(-1) range, for six resolutions, with a conventional blackbody source and ensembles of differential spectra. The NESRs are well behaved and consistent with the expected dependence on resolution; however, they depend on source temperature at the highest (1 cm(-1)) and lowest (32 cm(-1)) resolutions, with little or no statistical dependence at intermediate resolutions. Residual source drift is shown to be the likely cause of the dependence at 1 cm(-1); the dependence on the source at 32 cm(-1) resolution is shown to be most probably due to photon noise. At intermediate resolutions the sensor noise is dominant. PMID- 15130019 TI - Measurement of the azimuthal dependence of cross-polarized lidar returns and its relation to optical depth. AB - We measure with a gated intensified CCD camera the cross-polarized backscattered light from a linearly polarized laser beam penetrating a cloud made of spherical particles. In accordance with previously published results we observe a clear azimuthal pattern in the recorded images. We show that the pattern is symmetrical, that it originates from second-order scattering, and that higher order scattering causes blurring that increases with optical depth. We also find that the contrast in the symmetrical features can be related to measurement of the optical depth. Moreover, when the blurring contributions are identified and subtracted, the resulting pattern provides a pure second-order scattering measurement that can be used for retrieval of droplet size. PMID- 15130020 TI - Effects of sample temperature in femtosecond single-pulse laser-induced breakdown spectroscopy. AB - As much as tenfold atomic emission enhancements have been observed in experiments combining nanosecond (ns) and femtosecond (fs) laser pulses in an orthogonal dual pulse configuration for laser-induced breakdown spectroscopy (ns-fs orthogonal dual-pulse LIBS). In the examination of one of several potential sources of these atomic emission enhancements (sample heating by a ns air spark), minor reductions in atomic emission and as much as 15-fold improvements in mass removal have been observed for fs single-pulse LIBS of heated brass and aluminum samples. These results suggest that, although material removal with a high-powered, ultrashort fs pulse is temperature dependent, sample heating by the ns air spark is not the source of the atomic emission enhancements observed in ns-fs orthogonal dual pulse LIBS. PMID- 15130021 TI - Parametric study of pellets for elemental analysis with laser-induced breakdown spectroscopy. AB - The effect of various parameters on the accuracy of the laser-induced breakdown spectroscopy (LIBS) data taken from pellet samples has been investigated. The dependence of the standard deviation of the LIBS data on the amount and nature of the binder used, pressure used to press the powder into a pellet, and the position of the focal spot on the pellet has been investigated. Pellets made from industrially important materials such as silica, alumina, and lime with polyvinyl alcohol, sucrose, and starch as binders have been studied. The results thus obtained are tested by preparation of the calibration curves for Si, Fe, and B in the pellets made from the powder glass batch used as a surrogate for the batch employed for the vitrification of radioactive waste. PMID- 15130022 TI - Assessing reproducibility for interval data in health-related quality of life questionnaires: which coefficient should be used? AB - In contrast to other reliability estimates, test-retest reliability (or reproducibility) captures not only the measurement error of an assessment instrument, but also the stability of the construct measured. Consequently, one would expect any departure from identity (Y = X) of measurement pairs (X first, and Y second measurement) to be treated as 'error' by the respective reproducibility statistic, even if 'true' changes happened, e.g. worsening of a disease due to its natural course. The Pearson correlation, still often advocated for continuous measures in test-retest reliability studies, however captures the degree of linearity (Y = bX + a): perfect relationship can be computed, even if the measurement pairs differ not only by a additive constant 'a', but also because of a multiplication of the X-values with the slope 'b'. Therefore, intraclass correlation coefficients (ICCs) have been proposed as alternative statistics for reproducibility. However, only ICCs with absolute agreement definition of concordance capture the degree of identity. ICCs with a consistency definition of concordance measure the degree of additivity (Y = X + a). ICCs are calculated from repeated measures analyses of variance (ANOVAs), and a common population variance must be is assumed for the different measurements. Given this assumption, an ICC computed from a one-way ANOVA seems to be the best choice for this purpose. Otherwise, Lin's concordance correlation coefficient is recommended as identity measure. PMID- 15130023 TI - Assessing relationships between health-related quality of life and adherence to antiretroviral therapy. AB - OBJECTIVE: To investigate associations between health-related quality of life (HRQoL), as assessed using the multidimensional quality of life-HIV (MQOL-HIV) questionnaire, and adherence to antiretroviral treatment in HIV-infected subjects. DESIGN: Multicentre cross-sectional study in three institutional tertiary hospitals in northwest Spain. PATIENTS AND METHODS: The MQOL-HIV was completed by 235 HIV-infected adults undergoing antiretroviral treatment. Adherence to antiretroviral therapy was assessed by using patient's self-report. Information about sociodemographic characteristics and clinical variables was also collected. RESULTS: Good adherence (> or = 95% of prescribed pills correctly taken) was reported by 131 patients (55.7%). Univariate analyses indicated that the sociodemographic and clinical variables associated with adherence were age, educational level, income, employment, home stability, transmission route, history of previous antiretroviral therapy, and number of prescribed pills/day. Subscales of MQOL-HIV associated with adherence were mental health, cognitive functioning, financial status, medical care, partner intimacy, and (in men only) sexual functioning. Stepwise logistic regression showed that good adherence was more frequent in patients aged > 40 years (odds ratio, OR: 2.50; 95% confidence interval, CI: 1.15-5.61) and in patients with high cognitive functioning (OR: 2.26; 95% CI: 1.19-4.30). Conversely, poor adherence was more frequent in patients without stable home (OR: 2.96; 95% CI: 1.39-6.32), in patients required to take 14 or more pills/day (OR: 2.17; 95% CI: 1.18-4.28), in patients with low financial status (OR: 3.42; 95% CI: 1.57-7.45), and in patients reporting low medical care (OR: 2.07; 95% CI: 1.07-3.98). CONCLUSIONS: HRQoL dimensions, notably cognitive functioning, financial status and medical care, are closely associated with antiretroviral therapy adherence. PMID- 15130024 TI - Quality of life among people living with HIV/AIDS in northern Thailand: MOS-HIV Health Survey. AB - OBJECTIVES: Translation and psychometric evaluation of a Thai version of the Medical Outcomes Study HIV Health Survey (MOS-HIV) in Thailand. METHODS: A cross sectional survey in Chiang Mai province, northern Thailand, with data collected in face-to-face interviews using a structured questionnaire designed to measure 10 scales of quality of life (QOL). We recruited 200 people with HIV/AIDS attending self-help groups in the municipal area. Standard guidelines were followed for questionnaire translation and psychometric evaluations. RESULTS: Item-level internal consistency and discriminant validity were reasonably established. Success rates were 93.8 and 97.4%, respectively. Scale-level internal consistency reliability of multi-item scales was satisfactory, ranging from 0.74 to 0.88, with all exceeding inter-scale correlations. Principal components analysis of item and scale scores identified two hypothesized dimensions of the MOS-HIV. The mental health component was strongly loaded by health distress, mental health, vitality and cognitive function scales, and physical health by role, physical and social functions, and pain scales. Respondents manifesting symptoms or reporting worsening health status scored significantly lower on all scales. CONCLUSIONS: These preliminary studies have shown the Thai version of the MOS-HIV to have psychometric properties comparable with those reported in previous surveys. Further testing and modification should make it useful as an HIV-specific QOL measure in Thailand. PMID- 15130025 TI - Finding a useful conceptual basis for enhancing the quality of life of nursing home residents. AB - In this article it is depicted that before nursing home staff can effectively contribute to optimising the quality of life (QOL) of nursing home residents, it has to be clear what exactly QOL is and how it can be enhanced. The aim is to identify a QOL framework that provides tools for optimising QOL and can form the basis for the development of guidelines for QOL enhancement. For that purpose, a framework should meet three basic criteria: (1) it should be based on assumptions about comprehensive QOL of human beings in general; (2) it should clearly describe the contribution of each dimension to QOL and identify relationships between the dimensions; (3) it should take individual preferences into account. After the criteria are defined, frameworks identified from a literature search are discussed and evaluated according to these criteria. The most suitable framework appears to be the QOL framework of the theory of Social Production Functions. The implications of this framework in understanding the QOL of nursing home residents are described and recommendations for further research are discussed. PMID- 15130026 TI - Toward a model of quality of life for family caregivers of stroke survivors. AB - The important role of family caregivers in maintaining their disabled and elderly members in the community is becoming increasingly recognized. Caregiver research, for the most part, has explored burden and emotional distress as outcomes of the caregiving experience. Although there is a growing consensus among health-care researchers concerning the importance of quality of life (QoL) as an outcome, there is little research examining QoL of family caregivers. The purpose of this paper, therefore, was to construct a conceptual framework from which to study the QoL of family caregivers of stroke survivors. Findings from a review of studies addressing the QoL of these caregivers guided the development of the model. The components of the model include the caregiving situation, characteristics of the caregiver, and environmental factors, and their proposed relationships with QoL. This model provides a framework for investigating how the caregiving experience impacts on the caregiver's QoL. PMID- 15130027 TI - Quality of life in tuberculosis: patient and provider perspectives. AB - Tuberculosis (TB) is a persistent problem in the United States; however, little is known about its impact on functioning and quality of life (QOL) among people with TB. The purpose of this study is to describe the impact of TB on patients' QOL by using focus groups to assess the domains of QOL that are affected. Participants included patients (n = 10) who received treatment for active TB and physicians (n = 4) and nurses (n = 9) caring for patients with TB at a public health clinic in Baltimore, Maryland. TB affected all predicted domains of QOL, including general health perceptions, somatic sensation, psychological health, spiritual well-being, and physical, social and role functioning. Social stigmatization, isolation, pill burden, long duration of therapy, sexual dysfunction, loss of income, and fear were additional specific problems related to TB. Surprisingly, 11% (33) of the comments described benefits of TB illness, including increased spirituality and improved life perspectives. In addition, four additional QOL domains and three elements of treatment specific to TB which substantially impact QOL were identified. While patients and clinicians both identified issues in many areas of QOL, only patients mentioned the impact on sexual function, spirituality and improved life perspectives. Despite available curative therapy, TB and its treatment still have significant short and long-term consequences on patients' QOL. PMID- 15130028 TI - Feasibility and reliability of health-related quality of life measurements among tuberculosis patients. AB - The dramatic global impact of tuberculosis on mortality has been well documented, but its impact on morbidity has not been well described. The emphasis on treatment of latent tuberculosis (TB) infection highlights the tradeoff between short-term decrements in health status from 'preventive' therapy, and long-term gains related to fewer cases of active TB. However, these changes in health status have not been characterized. As a first step, we examined the feasibility and reliability of administering two health status questionnaires, in a multicultural TB clinic setting. The Medical Outcomes Study SF-36 and the EuroQOL EQ-5D were self-administered during 3 weekly interviews. One hundred and eighty six potentially eligible patients were identified, of whom 112 could be evaluated; 106 (57%) were confirmed eligible. Sixty-seven (63%) agreed to participate; 24 (36%) were women. Fifty-three participants (79%) were foreign born, with median residence in Canada of 3.5 years. Fifty (75%) of the participants completed all study measurements: 25 were treated for latent TB, 17 for active TB, and eight had previous active TB. Cronbach's alpha coefficients ranged from 0.73 to 0.94 for the SF-36 domain scores. Intraclass correlation coefficients were 0.66 for the SF-36 physical component summary, 0.79 for the mental component summary, and 0.73 for the EQ-5D. These instruments appeared reliable in a highly selected group of TB patients. PMID- 15130029 TI - Leisure time physical activity and health-related quality of life: cross sectional and longitudinal associations. AB - Studies that relate change in physical activity to change in health-related quality of life in the general population are needed to confirm associations suggested by cross-sectional studies. In the present study, cross-sectional as well as longitudinal associations between leisure time physical activity and health-related quality of life were studied in an apparently healthy population. The present study showed cross-sectional associations between at least moderately intense leisure time physical activity and general health perceptions, vitality, physical functioning and role limitations due to physical health problems. No associations were present for total leisure time physical activity. Change in leisure time physical activity was associated with change in social functioning in men as well as in women, irrespective of the intensity of physical activity. Only in men, change in total leisure time physical activity was associated with change in vitality and general mental health. In our study, cross-sectional associations were not confirmed by longitudinal analyses. Cross-sectional associations were mainly found for physical components of health-related quality of life, whereas longitudinal associations were predominantly observed for mental components of health-related quality of life. Confirmation of our results by those of other studies is needed in order to quantify health promotion messages. PMID- 15130030 TI - Validation of the Restless Legs Syndrome Quality of Life Instrument (RLS-QLI): findings of a consortium of national experts and the RLS Foundation. AB - This study was designed to assess the initial psychometric properties of a new disease-specific health-related quality of life (HRQL) measure, the Restless Legs Syndrome (RLS) Quality of Life Instrument (RLS-QLI). METHODS: Draft items were generated from a literature review, consultation with MD and PhD specialists in the fields of neurology and sleep medicine, and input from two patient focus groups. The initial item reduction was accomplished using a survey of 392 persons with self-reported RLS symptoms from the membership of the RLS Foundation. The final (independent) validation sample consisted of 574 of persons on the RLS Foundation's Interest Group List Serve who also reported having RLS. The mean age of participants was 54.5 (SD 12.3), with a sex ratio of 1M:2F, and the majority was on some form of medication for RLS (66%). RESULTS: Four factors were identified (Daily Function, Social Function, Sleep Quality, and Emotional Well Being) consisting of 17 items that explained 73.3% of the total variance. Each scale had good internal consistency (Cronbach's alpha's between 0.85 and 0.91) and 2-week test retest stability (Pearson Correlations between 0.81 and 0.93). Convergent validity was demonstrated using related scales on the SF-36 (r = 0.47 0.60) and criterion-related validity was shown using the clinical IRLS Scale of Symptom Severity (r = -0.45 to -0.77). CONCLUSION: The RLS-QLI is a valid disease specific HRQL instrument that will contribute to our understanding of how RLS impacts the lives of those affected with this CNS disorder. PMID- 15130031 TI - Development of the Short Form Endometriosis Health Profile Questionnaire: the EHP 5. AB - OBJECTIVE: To develop and validate a short form version of the Endometriosis Health Profile-30 questionnaire which consists of a 30-item core questionnaire and a 23 item modular questionnaire. METHODS: Three studies were carried out to develop the Endometriosis Health Profile-5. Study 1: a short form version of the Endometriosis Health Profile-30 core questionnaire was developed. Study 2: the results were verified in a new data set. Study 3: a short form version of the 23 item modular questionnaire was produced. The modular questionnaire contains six dimensions which may not be applicable to every woman with endometriosis and is used to supplement the five scales on the core questionnaire when required. RESULTS: The final instrument contained 11 items: five items from the core questionnaire and six items from the modular questionnaire. The 95% confidence intervals for Study 1 indicated that the scale scores overlapped with each item score and were confirmed in the second analysis (study 2). Each item was most highly correlated to its parent scale and less with the remaining four core questionnaire scales which was verified in study 2. In both studies all correlations were significant at the 0.01 level (two tailed test). CONCLUSION: The Endometriosis Health Profile-5 is a reliable and valid short form questionnaire which can also be supplemented with a short form version of the modular questionnaire when required. It will be especially useful in clinical settings where a short and economical endometriosis health status measure is required. PMID- 15130033 TI - Early root alterations after orthodontic force application studied by light and scanning electron microscopy. AB - The purpose of the study was to characterize root surface alterations in orthodontically moved teeth. Thirty-six 40-50-day-old male Wistar rats were used. The maxillary right first molar was mesialized by means of a fixed appliance, exerting 50 g of force upon insertion. One, 2 and 4 days after force application the animals were sacrificed (nine animals per observation period) and block sections processed for analysis. Nine animals served as untreated controls. In total, 20 specimens were prepared for examination of the mesial aspect of the mesiobuccal root by scanning electron microscopy (SEM). The remaining specimens were processed for light microscopy. Three morphologically distinct types of resorption defect were observed: isolated small lacunae, wide shallow resorption bays, and deep resorption lacunae. The area occupied by each resorption type varied significantly (P < 0.05) with time. Isolated small lacunae were the earliest to be observed, sometimes found in continuity with wide shallow resorption bays. Mononucleated macrophage-like cells were associated with both resorption types. Deeper lacunae extending into the dentine were found at the 4 day observation period. These always occurred within shallow resorption bays and appeared to be created by multinucleated cells. From the sequence of the different root surface defects and associated cell types observed during the development of orthodontically induced root resorption, it may be concluded that different cell types, with different resorptive potential but functionally interrelated, are involved in the successive phases of the process, and that each cell type leaves a characteristic resorption pattern on the root surface. PMID- 15130032 TI - Evaluating the responsiveness of the Endometriosis Health Profile Questionnaire: the EHP-30. AB - OBJECTIVES: To evaluate the sensitivity to change of the Endometriosis Health Profile-30 (EHP-30) questionnaire. SETTING: The Women's Center, John Radcliffe Hospital, Oxford. DESIGN: Postal survey to 66 women undergoing conservative surgery for the treatment of endometriosis-associated pain. The EHP-30 and the Short Form-36 (SF-36) were administered 2 weeks before the operation, and 4 months post-operatively. At T2 a transition question was included to evaluate changes in patients health status. To evaluate responsiveness effect sizes, standardised response means, the index of responsiveness and the minimally and clinically important differences were calculated. RESULTS: Forty (66.6%) patients returned the questionnaires at time 1 and 2. Overall less responsive effect size scores were found for the SF-36 (0.1-0.5) compared to the EHP-30 (-0.1-1.1) for all patients who had undergone treatment. Minimally important differences and the index of responsiveness were overall higher for the EHP-30 (0.4-2.0) compared to the SF-36 (0.1-1.0). Change scores for four of the five scales were significantly correlated with women's responses to the transition question. CONCLUSIONS: Results suggest that the EHP-30 is sensitive to change. Its application in clinical trials should prove beneficial in assessing the impact of medical and surgical interventions upon quality of life for women with endometriosis. PMID- 15130034 TI - In vitro expression of matrix metalloproteinase-1, tissue inhibitor of metalloproteinase-1 and transforming growth factor-beta1 in human periodontal ligament fibroblasts. AB - Extracellular matrix remodelling is mediated via matrix metalloproteinases (MMPs) and their regulatory factors such as tissue inhibitors of metalloproteinases (TIMPs) and transforming growth factor-beta (TGF-beta). The regulation of MMPs is thought to be associated with cytoskeletal changes. In this study, cytoskeletal changes in human periodontal ligament fibroblasts (PDFs) were induced using cytochalasin B (CB) which reorganizes actin microfilaments reversibly, and colchicine which disrupts microtubules irreversibly. The levels of MMP-1, TIMP-1 and TGF-beta secreted by the CB- or colchicine-treated PDFs were measured using an enzyme-linked immunosorbent assay. Differences between experimental and control groups were tested using analysis of variance (ANOVA) and Scheffe's ad hoc test. Although CB treatment did not significantly increase MMP-1 expression over the controls, colchicine treatment significantly increased the expression of MMP-1 (P < 0.01) in a time-dependent manner compared with the controls. Both CB and colchicine showed a time-dependent increase in TIMP-1 and TGF-beta1 expression and a dose-dependent increase in TIMP-1 and TGF-beta1 expression until threshold compared with the controls (P < 0.05, P < 0.01 and P < 0.001). In addition, CB treatment produced significantly increased TGF-beta1 expression over the controls (P < 0.05 and P < 0.001) from lower doses, with this effect occurring at earlier time points compared with colchicine treatment. PMID- 15130035 TI - Neovascularization and bone formation in the condyle during stepwise mandibular advancement. AB - The aims of this investigation were to identify the temporal expression of vascular endothelial growth factor (VEGF) in the mandibular condyle and to correlate it with the pattern of new bone formation during stepwise mandibular advancement. Two hundred and fifty female, 35-day-old Sprague-Dawley rats were randomly divided into 10 groups, with 10 rats allocated to the single-step bite jumping subgroup, 10 rats to the stepwise advancement subgroup and five rats to the control subgroup. In the experimental groups, the mandibles were kept in a continuous forward position. The initial stepwise advancement commenced on day 35, whereas the second advancement started on day 65. The rats were sacrificed on experimental days 3, 7, 14, 21, 30, 33, 37, 44, 51 and 60. Sections (7 microm) were cut through the condyle in the parasagittal plane and stained with anti-VEGF antibody. Each section was counter-stained with haematoxylin for observation of the cellular response. The sections were digitized and quantitatively analysed with a computer-assisted image analysing system. The results showed that the initial advancement in the stepwise group led to significantly less expression of VEGF when compared with single advancement. However, the second advancement on day 30 resulted in a significant increase in VEGF expression when compared with the one-step group and the natural growth control group. Thus, it was concluded that changes in the amplitude of mechanical loading, produced by stepwise advancement, have a significant effect on the production of VEGF by the chondrocytes. During the later stages of advancement, more VEGF and more condylar bone was produced. PMID- 15130036 TI - Secretion of osteopontin from MG-63 cells under a physiological level of mechanical strain in vitro--a [35S] incorporation approach. AB - To gain insight into the early response of osteoblastic cells to a physiological level of mechanical strain in vitro, the secretion of osteopontin by MG-63 osteosarcoma cells was assessed by [35S] incorporation and autoradiography. First, osteopontin secreted from MG-63 cells was immunolocalized at 60-64 kDa (Mr) by polyacrylamide gel electrophoresis. A uniform physiological level of strain was generated by a vacuum added to the convex side of a half-ball shaped silicon rubber membrane on which the cells were cultured on the concave side. After labelling proteins with [35S]-methionine/cysteine (147 microCi/ml), the membranes were exposed to a strain of 0.5 per cent (5000 microepsilon), 3 cycles/minute (sine wave) with 10 minutes on and off. At 1, 2 and 4 hours after strain, the supernatants were collected and analysed by 10 per cent sodium dodecyl sulphate-polyacrylamide gel electrophoresis and autoradiography. The results showed that osteopontin was secreted by the strained cells at significantly higher amounts than the non-strained cells at all three time points (P < 0.05), with the first hour being the most prominent. A physiological level of mechanical strain increased the secretion of osteopontin from MG-63 cells in an early phase. This finding implies an accelerated process of bone remodelling, which suggests that the application of light and intermittent forces would result in the cellular reaction identified in relation to orthodontic tooth movement. The results indirectly indicate that the level of force presently used might be too high. PMID- 15130037 TI - Dental crowding in a prehistoric population. AB - The aims of this study were to investigate dental crowding from the Copper Age and examine the extent and patterns of wear. Crowding was estimated in 43 adult mandibles using Little's irregularity index. Dental wear, dental diameters, arch width and the presence of third molars were also studied as possible aetiological factors for crowding. The remains were found at the archaeological site of Roaix, located in the south of France. Radiocarbon dating indicated that the lower layer was from 2150 +/- 140 years BC (date +/- 1 standard deviation) and the upper level from 2090 +/- 140 years. The graves were estimated to contain the remains of 150 adults and 50 children. Forty-three intact mandibles were used for this study. All of the mandibles presented incisor crowding with a majority of minimal and moderate irregularities, but in seven cases there were extreme irregularities and in two canine impaction was observed. These results are in contrast with the literature where it is reported that malocclusions were rare in prehistoric populations. The findings of this study suggest that crowding may be of a genetic origin and might not be caused by excessive tooth size or changes in environmental factors (masticatory activity). PMID- 15130038 TI - Tensile properties of orthodontic elastomeric chains. AB - The purpose of this investigation was to assess the permanent elongation, tensile strength and toughness of elastomeric chains. Two types (open and closed chains, i.e. with and without an intermodular link) of three brands of elastomeric module yielding six groups were included in the study. Specimens with equal numbers of loops were measured and classified into four groups based on their ageing state: (a) as-received; (b) subjected to a 24 hour steady strain in air determined as 50 per cent of original length; (c) exposed intraorally for 24 hours; and (d) retrieved following 3 weeks of intraoral exposure. All specimens were subjected to tensile stress and their behaviour was analysed with three-way ANOVA and Tukey's multiple comparison test at alpha = 0.05 level of significance, with brand, type (open or closed chains), and treatment variables serving as discriminating variables. Modular geometry or design were found not to have significant effects on elongation, probably due to the substantial variation in chain shape, modular size, and link length among products of the same category. Similarly, no correlation was identified between specimen treatment and the tensile strength of elastomers. The toughness results were not consistent with the group rankings for tensile strength, probably because of variation in the elastic and plastic deformation of specimens upon loading. PMID- 15130039 TI - Dynamic frictional behaviour of orthodontic archwires and brackets. AB - The aim of this study was to evaluate the frictional behaviour of 15 different archwires and 16 different brackets using small oscillating displacements when opposed to a standard stainless steel bracket or a standard stainless steel wire. Tests were run according to a pilot study at a frequency of 1 Hz and with a reciprocating tangential displacement of 200 microm, while the wire remained centred in the bracket slot under a load of 2 N. The results indicated a significant difference between the evaluated wires and brackets. The mean coefficient of friction (COF) of the wires varied from 0.16 for Imagination NiTi tooth-coloured wire to 0.69 for the True Chrome Resilient Purple wire, while for the brackets it ranged from 0.39 for Ultratrimm to 0.72 for the Master Series. The fact that in this study, a large number of different commercially available archwires and brackets were evaluated with the same apparatus according to the same protocol, allows a direct comparison of the different archwire and bracket combinations, and can assist in the choice of the optimal bracket-wire combination with regard to friction. PMID- 15130040 TI - Metal ion release from new and recycled stainless steel brackets. AB - As orthodontic appliances can corrode with time in the oral environment, the aim of this study was to compare the release of metal ions from new and recycled brackets immersed in buffers of different pH values over a 48 week period. To simulate commercial recycling, the stainless steel brackets were divided into two groups: new and recycled. The bases of the latter were coated with adhesive and the brackets were heat treated before being immersed in the test solution for 48 weeks. The release of nickel, chromium, iron, copper, cobalt and manganese ions was analysed by atomic absorption spectrophotometry. Differences were compared using one-way analysis of variance. The results showed that recycled brackets released more ions than new brackets (P < 0.05). Brackets immersed in solutions of pH 4 released more ions than those immersed in solutions of pH 7, and the total amount of ions released increased with time over the 48 week period (P < 0.05). This study demonstrates that both new and recycled brackets will corrode in the oral environment. To avoid clinical side-effects, metal brackets should be made more resistant to corrosion, and recycled brackets should not be used. PMID- 15130041 TI - Evaluation of shear bond strength with different enamel pre-treatments. AB - The purpose of this study was to investigate the shear bond strengths of two adhesives, Panavia-21 and a composite resin (Transbond XT), with different enamel pre-treatments, acid etching (37 per cent phosphoric acid) and grit blasting (50 microm aluminium oxide particles). The mode of bond failure was also assessed using the modified adhesive remnant index (ARI). Ninety freshly extracted non carious human premolar teeth were randomly divided into the following groups: (1) Transbond XT, acid-etched enamel surface; (2) Panavia-21, acid-etched enamel surface; (3) Transbond XT, grit-blasted enamel surface; (4) Panavia-21, grit blasted enamel surface; (5) Transbond XT, acid-etched enamel surface with grit blasted brackets; (6) Panavia-21, acid-etched enamel surface with grit-blasted brackets. All groups had stainless steel brackets bonded to the buccal surface of each tooth. An Instron universal testing machine was used to determine the shear bond strengths at a crosshead speed of 0.5 mm/second. Statistical analysis was undertaken using analysis of variance and the Tukey test. The mean bond strength values were as follows: group 1, 135.7 +/- 23.0 N; group 2, 181.5 +/- 18.4 N; group 3, 38.4 +/- 27.5 N; group 4, 59.1 +/- 24.1 N; group 5, 106.7 +/- 21.5 N; group 6, 165.3 +/- 21.4 N. Panavia-21 with the acid-etched enamel surface had a significantly higher shear bond strength than the other groups (P < 0.001). This was followed by the composite group with the acid-etched enamel surface. This group differed significantly from the composite and Panavia-21 groups with the grit-blasted tooth surface (P < 0.001) and from the composite and Panavia-21 groups with the acid-etched enamel surface and grit-blasted brackets (P < 0.01). The current findings indicate that Panavia-21 is an excellent adhesive and produces a bond strength that is clinically useful. Enamel surface preparation using grit blasting alone results in a significantly lower bond strength and should not be advocated for clinical use. PMID- 15130042 TI - A clinical study of glass ionomer cement. AB - The aim of this investigation was to compare the clinical performance of a glass ionomer cement (GIC) with a composite resin when used for direct bonding of standard edgewise orthodontic brackets. Fourteen patients (10 females, four males), in whom 242 teeth were bonded with brackets, were divided into two groups: GIC (121 teeth) and composite (121 teeth). The brackets were allocated to alternate quadrants and first-time failures were recorded over a period of 24 months. Data were analysed statistically (non-parametric chi-squared test). The results demonstrated a significantly lower unpreviewed debonding index (UDI) (15.7 per cent) for the composite than for the GIC (28.1 per cent) (P = 0.042). The use of heavy archwires was largely responsible for this difference. No difference was observed when light and medium archwires were used. GIC may be a viable alternative to composite for use with light archwires and with limited treatment objectives. PMID- 15130043 TI - An orthopaedic approach to the treatment of Class III malocclusions in the early mixed dentition. AB - The aim of this investigation was to study cephalometrically the skeletal, dental and soft tissue modifications induced by a Delaire facemask and Bionator III appliance in a sample of 30 patients (17 boys and 13 girls), aged 4.1-9 years [mean 5.85 years, confidence interval (CI) 5.41-6.29], in the early mixed dentition with a skeletal Class III malocclusion caused by maxillary retrognathism (group 1) and compared with a control sample of 24 subjects (14 boys and 10 girls), aged 4-9 years (mean 5.97 years, CI 5.35-6.58) with untreated Class III malocclusions (group 2). For each patient a lateral cephalogram was taken before treatment (T0), after facemask removal (T1), and at the end of the retention period with a Bionator III (T2). Cephalometric analysis was carried out. The post-treatment cephalometric values in the treated group showed a forward displacement of the maxilla resulting in a statistically significant increase (P < 0.001) in the SNA angle, A-NPg (mm) and PNS-A (mm) linear values. There was a clockwise rotation of the mandible, with a decrease in the SNB angle and a satisfactory correction of the Class III relationship. The beneficial effects on the facial profile were confirmed by an increase in UL-EL distance and in NB--HL and NsPgs--HL angles, and by a decrease in the facial convexity angle. These findings indicate that the Delaire facemask and Bionator III treatment is effective for correcting skeletal Class III malocclusions caused by maxillary retrognathism in the early mixed dentition. PMID- 15130044 TI - Variation in the cranial base orientation and facial skeleton in dry skulls sampled from three major populations. AB - The aim of this study was to analyse the effects of cranial base orientation on the morphology of the craniofacial system in human populations. Three geographically distant populations from Europe (72), Africa (48) and Asia (24) were chosen. Five angular and two linear variables from the cranial base component and six angular and six linear variables from the facial component based on two reference lines of the vertical posterior maxillary and Frankfort horizontal planes were measured. The European sample presented dolichofacial individuals with a larger face height and a smaller face depth derived from a raised cranial base and facial cranium orientation which tended to be similar to the Asian sample. The African sample presented brachyfacial individuals with a reduced face height and a larger face depth as a result of a lowered cranial base and facial cranium orientation. The Asian sample presented dolichofacial individuals with a larger face height and depth due to a raised cranial base and facial cranium orientation. The findings of this study suggest that cranial base orientation and posterior cranial base length appear to be valid discriminating factors between different human populations. PMID- 15130045 TI - Quantitation of transverse maxillary dimensions using computed tomography: a methodological and reproducibility study. AB - The present investigation was a methodological study of a new method of quantification of a series of factors in the transverse dimension of the maxilla including the nose, maxillary bones and dental arches, based on computer tomographic (CT) scanning. The aim was to investigate a series of parameters thought to be relevant in the differential diagnosis of discrepancies in the morphology of this area and probably affected by orthodontic appliances. Based on a standardized CT scanning registration of 10 subjects, a series of points on the scans were identified and then measured in a special cephalometric computer system (linear and angular values). The quantitation was repeated by each observer and inter- and intra-observer differences were calculated. The results demonstrated that virtually all the parameters showed a high degree of reproducibility at both levels and confirmed the statistical suitability of the method described. The method will be used in a series of ongoing studies regarding the morphology and treatment of discrepancies of the midface and therefore supplement the relatively sparse information based on quantitative reports concerning this important anatomical area. PMID- 15130046 TI - A preliminary assessment of cephalometric orthodontic superimposition. AB - The superimposition of cephalograms is a valuable tool in orthodontics. In children this task is complicated by the fact that growth changes in the reference structures used in superimposition should be taken into consideration. In this study a new method of superimposition of cephalograms taken in the natural head position (NHP) was compared with Viazis' cranial base triangle and a reference grid was used to quantitatively assess the changes in the five selected landmarks. The material consisted of 12 pairs of cephalograms of growing subjects (mean age 11.1 years) collected with a time interval of 1 year. The results revealed no significant statistical difference between the two methods and both showed high reproducibility. Both analyses were found to be suitable for individual assessment, but the new method involving the use of NHP and easy landmark identification can be considered as a useful addition. PMID- 15130047 TI - Effects of bilateral upper first premolar extraction on the mandible. AB - The purpose of this study was to evaluate the effects of bilateral upper premolar extraction on mandibular growth. Twenty-six subjects (eight males, 18 females) in maximum pubertal growth with an Angle Class II molar relationship, normal to mild overjet increase, mild or no lower arch length discrepancy but severe upper arch discrepancy and no severe skeletal discrepancy were divided into two groups equal in number and gender, as extraction and control groups. The median chronological age was 11.2 years in the extraction group and 12.6 years in the controls. The subjects were observed for a median period of 1.1 years in the extraction group after bilateral extraction of the upper premolars and 1.2 years in the controls until termination of pubertal growth (DP3u) without any orthodontic treatment. Twenty-nine linear and angular measurements were made on 52 lateral cephalograms and hand-wrist radiographs taken before and after the study period. The increase in SNB measured on the total superimposition was significantly greater in the controls than in the extraction group (P < 0.05). In addition, anterior mandibular (counter-clockwise) rotation was only significant (P < 0.05) in the control group. Thus, it might be suggested that bilateral upper premolar extractions might affect the mandibular rotation tendency. PMID- 15130048 TI - Tests for comparing mark-specific hazards and cumulative incidence functions. AB - It is of interest in some applications to determine whether there is a relationship between a hazard rate function (or a cumulative incidence function) and a mark variable which is only observed at uncensored failure times. We develop nonparametric tests for this problem when the mark variable is continuous. Tests are developed for the null hypothesis that the mark-specific hazard rate is independent of the mark versus ordered and two-sided alternatives expressed in terms of mark-specific hazard functions and mark-specific cumulative incidence functions. The test statistics are based on functionals of a bivariate test process equal to a weighted average of differences between a Nelson-Aalen type estimator of the mark-specific cumulative hazard function and a nonparametric estimator of this function under the null hypothesis. The weight function in the test process can be chosen so that the test statistics are asymptotically distribution-free. Asymptotically correct critical values are obtained through a simple simulation procedure. The testing procedures are shown to perform well in numerical studies, and are illustrated with an AIDS clinical trial example. Specifically, the tests are used to assess if the instantaneous or absolute risk of treatment failure depends on the amount of accumulation of drug resistance mutations in a subject's HIV virus. This assessment helps guide development of anti-HIV therapies that surmount the problem of drug resistance. PMID- 15130049 TI - Modeling event times with multiple outcomes using the Wiener process with drift. AB - Length of stay in hospital (LOS) is a widely used outcome measure in Health Services research, often acting as a surrogate for resource consumption or as a measure of efficiency. The distribution of LOS is typically highly skewed, with a few large observations. An interesting feature is the presence of multiple outcomes (e.g. healthy discharge, death in hospital, transfer to another institution). Health Services researchers are interested in modeling the dependence of LOS on covariates, often using administrative data collected for other purposes, such as calculating fees for doctors. Even after all available covariates have been included in the model, unexplained heterogeneity usually remains. In this article, we develop a parametric regression model for LOS that addresses these features. The model is based on the time, T, that a Wiener process with drift (representing an unobserved health level process) hits one of two barriers, one representing healthy discharge and the other death in hospital. Our approach to analyzing event times has many parallels with competing risks analysis (Kalbfleisch and Prentice, The Statistical Analysis of Failure Time Data, New York: John Wiley and Sons, 1980)), and can be seen as a way of formalizing a competing risks situation. The density of T is an infinite series, and we outline a proof that the density and its derivatives are absolutely and uniformly convergent, and regularity conditions are satisfied. Expressions for the expected value of T, the conditional expectation of T given outcome, and the probability of each outcome are available in terms of model parameters. The proposed regression model uses an approximation to the density formed by truncating the series, and its parameters are estimated by maximum likelihood. An extension to allow a third outcome (e.g. transfers out of hospital) is discussed, as well as a mixture model that addresses the issue of unexplained heterogeneity. The model is illustrated using administrative data. PMID- 15130050 TI - Incorporating follow-up time in M-estimation for survival data. AB - It has been approximately 30 years since D.R. Cox introduced the proportional hazards method to model the relationship between covariates and survival time. However, the proportional hazards model has limited value when the proportionality assumption is violated. Over the years, there have many been many alternative proposals to the proportional hazards regression model for the case of right censored survival data, but to date none have demonstrated widespread acceptance. In general, problems encountered in these methods include their computational algorithms or evaluation of their asymptotic properties. In this work, an estimating equation based on a U-statistic of degree 2 is proposed. It is easy to implement and the U-statistic framework provides a straightforward development of asymptotic inferential theory for the regression parameters. PMID- 15130051 TI - Statistical analysis of linear degradation and failure time data with multiple failure modes. AB - The paper considers linear degradation and failure time models with multiple failure modes. Dependence of traumatic failure intensities on the degradation level are included into the models. Estimators of traumatic event cumulative intensities, and of various reliability characteristics are proposed. Prediction of residual reliability characteristics given a degradation value at a given moment is discussed. Non-parametric, semiparametric and parametric estimation methods are given. Theorems on simultaneous asymptotic distribution of random functions characterising degradation and intensities of traumatic events are proposed. Asymptotic properties of unconditional and residual reliability characteristics estimators are given. Real tire wear and failure time data are analysed. PMID- 15130052 TI - Reliability estimation based on system data with an unknown load share rule. AB - We consider a multicomponent load-sharing system in which the failure rate of a given component depends on the set of working components at any given time. Such systems can arise in software reliability models and in multivariate failure-time models in biostatistics, for example. A load-share rule dictates how stress or load is redistributed to the surviving components after a component fails within the system. In this paper, we assume the load share rule is unknown and derive methods for statistical inference on load-share parameters based on maximum likelihood. Components with (individual) constant failure rates are observed in two environments: (1) the system load is distributed evenly among the working components, and (2) we assume only the load for each working component increases when other components in the system fail. Tests for these special load-share models are investigated. PMID- 15130053 TI - Growing interest in reptile behaviour. PMID- 15130054 TI - Risk of fatal distal limb fractures among Thoroughbreds involved in the five types of racing in the United Kingdom. AB - The risk of fatal distal limb fractures in thoroughbreds racing in the UK was calculated and shown to vary considerably between the different types of race. Flat turf racing was associated with the lowest risk (0.4 per 1000 starts) and national hunt flat racing was associated with the highest risk (2.2 per 1000 starts). The types of fracture were classified by detailed radiographic and postmortem examinations of all the cases recorded over two years, and the distribution of the different types of fracture in the five main types of racing was examined. Overall, lateral condylar fractures of the third metacarpus were the most common, and they were also the most common in national hunt-type races (hurdle, steeplechase and national hunt flat races). In all-weather flat racing biaxial proximal sesamoid fractures were most common, and in turf flat racing fractures of the first phalanx were most common. The risk of fractures of more than one bone was greater in national hunt-type races. PMID- 15130055 TI - Strains of avian paramyxovirus type 1 of low pathogenicity for chickens isolated from poultry and wild birds in Denmark. AB - Twenty-one strains of avian paramyxovirus type 1 of low virulence for chickens were isolated in Denmark between 1996 and the beginning of 2003. The low virulence of the strains was demonstrated by sequencing the fusion (F) gene at the cleavage site motif and in some cases by determining the intracerebral pathogenicity index in day-old chicks. By using a panel of monoclonal antibodies it was shown that the isolates belonged to four different antigenic groups (five C2 isolates, six E isolates, six H isolates and four G/Q isolates). They were placed in three distinguishable genetic groups by phylogenetic analysis of a partial sequence of the F gene. The origin of the six E isolates was probably contaminated vaccines; the other viruses were isolated from wild birds and from poultry which probably came into contact with wild birds. PMID- 15130056 TI - Changes in the feed intake, pH and osmolality of rumen fluid, and the position of the abomasum of eight dairy cows during a diet-induced left displacement of the abomasum. AB - During the last six weeks of the dry period, eight Holstein-Friesian cows were fed a restricted amount of grass silage; after calving, a mixture of maize silage and concentrates was offered in a feeding regimen designed to induce a displacement of the abomasum. In the first month after calving, the cows were monitored for the following variables: feed intake and composition, milk production, the position of the abomasum, and the pH and osmolality of the rumen contents. In five of the eight cows, a left displacement of the abomasum occurred between four and 21 days after calving in the absence of other diseases. The displacement was temporary, lasting between five and 36 consecutive hours and one or two days in two of the cows (floaters), and for three or more days in the other three. Before these three cows developed the displacement, their abomasum was 4.3 to 7.9 cm higher, its contents had a higher mean osmolality (+19.2 mosmol/kg), and the ratio of roughage to concentrates in their feed was lower ( 0.87) than in the three cows that did not develop clinical signs of a displaced abomasum. There were no significant differences in these variables between the floaters and the healthy cows. PMID- 15130057 TI - Survey of caseous lymphadenitis seroprevalence in British terminal sire sheep breeds. PMID- 15130058 TI - Escherichia coli O116 associated with an outbreak of calf diarrhoea. PMID- 15130059 TI - Extensive uroperitoneum and pleural effusion associated with necrotic urachal remnant in a bull calf. PMID- 15130060 TI - Questions regarding the professional training phase. PMID- 15130061 TI - Competition in the supply of medicines. PMID- 15130062 TI - Neurological complications in sheep following administration of parenteral copper. PMID- 15130063 TI - Discarded lead-acid batteries: a preventable cause of lead poisoning in cattle. PMID- 15130064 TI - Integrating the principles of evidence-based practice into clinical practice. AB - This series of articles illustrates many considerations relevant to the application of clinical practice guidelines (CPGs). This particular column describes the actions of a nurse practitioner (NP) striving to understand the foundations of recommendations that are based largely on expert opinion. Although application of CPGs does not generally require this degree of investigation, it is essential that providers understand the processes used to interpret the basis of recommendations, including the application of the basic statistical concepts, when making decisions about how recommendations apply to individual patient scenarios. Utilizing evidence-based practice when providing patient care requires a range of skills that allows the NP to locate appropriate research evidence, to develop an understanding of the statistics used in interpreting and reporting research, and to evaluate the effects of interventions on patient outcomes. The application of the key concepts of evidenced-based practice within the primary care setting is explored through a hypothetical patient scenario, which was created as the focal point for three articles that illustrate principles of evidence-based practice. The goal of this series of articles is to provide a basic understanding of evidence-based practice and its application in clinical practice. This article explores the use of interventions selected from CPGs and investigates the potential effects of recommended interventions on patient outcomes. Commonly encountered statistical concepts are reviewed, and examples of their application in interpreting and reporting research are demonstrated. The principles of relative risk, relative risk reduction, absolute risk reduction, and numbers needed to treat are described. This review provides the NP with some basic skills to determine both the quality and usefulness of research. PMID- 15130065 TI - The role of genetics in the risk of thromboembolism: prothrombin 20210A and oral contraceptive therapy. AB - PURPOSE: To provide an overview of the prothrombin 20210A mutation, its effects on the incidence of venous thromboembolism (VTE) in users of oral contraceptive therapy (OCT), and screening recommendations for the primary care practice setting. DATA SOURCES: Several databases, including MEDLINE, EMBASE, BIOSIS, Cochrane Library, and SciSearch, were searched for articles published between 1996 and 2003. An integrative review of studies addressing prothrombin 20210A was done using available case-series and case-control studies. No randomized controlled trials on prothrombin 20210A were available in the literature from the years searched. CONCLUSIONS: Prothrombin 20210A increases the risk of developing a VTE for those who carry the genetic mutation. The presence of either concomitant circumstantial factors (e.g., surgery or immobilization) or a combination of genetic factors (e.g., factor V Leiden and prothrombin 20210A) raises the frequency of VTEs to an even greater extent. The risk increases exponentially in users of OCT. Screening guidelines for the use of OCT in prothrombin 20210A carriers remain unclear due to the paucity of empirical evidence related to the topic. IMPLICATIONS FOR PRACTICE: Practitioners caring for a prothrombin 20210A carrier should maintain a high degree of suspicion with even vague signs or symptoms of a possible VTE. Practitioners should consider completing a full diagnostic workup for VTE on that patient, particularly if that patient is taking OCT. Until evidence becomes available as to the best anticoagulation practice after an initial or recurrent VTE in a prothrombin 20210A carrier, standard treatment guidelines for anticoagulation should be followed. PMID- 15130066 TI - A proposal for a code of ethics for nurse practitioners. AB - PURPOSE: To review established codes for health care professionals and standards of practice for the nurse practitioner (NP) and to utilize these codes and standards, general ethical themes, and a new ethical triangle to propose an ethical code for NPs. DATA SOURCES: Reviews of three generally accepted ethical themes (deontological, teleological, and areteological), the ethical triangle by Potter, the American Academy of Nurse Practitioners (AANP) standards of practice for NPs, and codes of ethics from the American Nurses Association (ANA) and the American Medical Association (AMA). CONCLUSIONS: A proposal for a code of ethics for NPs is presented. This code was determined by basic ethical themes and established codes for nursing, formulated by the ANA, and for physicians, formulated by the AMA. The proposal was also developed in consideration of the AANP standards of practice for NPs. IMPLICATIONS FOR PRACTICE: The role of the NP is unique in its ethical demands. The authors believe that the expanded practice of NPs presents ethical concerns that are not addressed by the ANA code and yet are relevant to nursing and therefore different than the ethical concerns of physicians. This proposal attempts to broaden NPs' perspective of the role that ethics should hold in their professional lives. PMID- 15130067 TI - Health promotion attitudes and practices of Texas nurse practitioners. AB - PURPOSE: The purpose of this descriptive cross-sectional survey was to examine the health promotion attitudes and practices of Texas nurse practitioners (NPs). DATA SOURCES: Original research utilizing the Health Promotion Practices of Nurse Practitioners Instrument developed by the researcher and completed by 442 Texas NPs. CONCLUSIONS: Overall, Texas NPs have positive attitudes toward health promotion and are supportive of health promotion practices. However, the findings of this study indicate that improvement can be made in the adoption of certain health promotion practices. IMPLICATIONS FOR PRACTICE: The growing number of NPs providing care to patients can significantly impact public health through consistent health promotion in clinical practice. PMID- 15130068 TI - Prospective analysis of presenting symptoms among 265 patients with radiographic evidence of Chiari malformation type I with or without syringomyelia. AB - OBJECTIVE: To characterize the presenting symptoms among persons with radiographic evidence of Chiari malformation (CM) type I in an effort to help the primary care provider determine accurate diagnosis and treatment. METHODS: The symptoms of CM can often be vague and ambiguous, leading to misdiagnosis. Patients who had radiographic evidence of CM with or without syringomyelia were prospectively studied. The detailed symptom data were collected from patient reports at initial examination to determine if there was a pattern of symptoms typically attributable to the CM. RESULTS: Analysis of presenting symptoms among 265 patients demonstrated a distinct pattern of symptom constellation. This study reports a prospective analysis of presenting symptoms among 265 patients with radiographic evidence of CM with or without syringomyelia and defines a common presentation pattern. Recognition of typical symptoms attributable to CM can result in appropriate diagnostic workup, accurate diagnosis, and timely treatment, leading to improved patient outcomes. PMID- 15130069 TI - Educational strategies to enhance placement and retention of nurse practitioners in rural Arkansas. AB - PURPOSE: To examine the factors and incentives enhancing nurse practitioners' (NPs') long-term employment in rural areas. DATA SOURCE: The participants were 121 master's prepared or higher educated NPs located in rural Arkansas. A survey about role preparation and employment factors was mailed to the participants. CONCLUSIONS: We found that 10% of all respondents were not currently employed in the role of an NP. The majority of rural-employed NPs had didactic content focusing on rural-practice opportunities and engaged in rural practicums while in graduate school. Rural NPs were much more likely to have graduated from research intensive universities. Nearly 90% of NPs reported they were somewhat to very well prepared for practice in rural Arkansas. IMPLICATIONS FOR EDUCATION: The shortage of health care providers remains one of the most irresolvable problems in the U.S. health care delivery system. Compounding the shortage is the unequal distribution of health care practitioners in rural areas. Evidence indicates that NP students engaged in rural clinical practicums were more likely to practice in rural areas. In light of this information, nurse educators should expand the NP educational process beyond rurally located practicums to deliver the entire educational process to the NP students' rural homes. PMID- 15130070 TI - The photographic revolution. PMID- 15130071 TI - Clinical comparison of a multistranded wire and a direct-bonded polyethylene reinforced resin composite used for lingual retention. PMID- 15130072 TI - Clinical comparison of a multistranded wire and a direct-bonded polyethylene reinforced resin composite used for lingual retention. PMID- 15130073 TI - Bonded amalgam sealants and adhesive resin sealants: five-year clinical results. AB - Bonded amalgams were used as pit-and-fissure sealants without mechanical preparation. They were compared with resin-based pit-and-fissure sealants for retention over a 5-year period. Clinical examinations at 6 months, 1 year, 2 years, and 5 years revealed no difference between the two techniques. Although amalgam sealants may not be practical by themselves, they can be used to seal pits and fissures surrounding very conservative preparations, in the "preventive amalgam restoration." Conventional amalgam retentive features and 90-degree cavosurface margins may not be necessary when bonding is used with amalgam. PMID- 15130074 TI - Direct posterior resin composite restorations: considerations on finishing/polishing. Clinical procedures. AB - In spite of advances in materials and techniques for direct posterior composite restorations, some problems remain, such as occlusal wear rate, marginal breakdown, and marked operator technique sensitivity. To enhance longevity of these restorations, finishing and polishing procedures are commonly indicated. Although these procedures offer advantages, they also have some drawbacks, such as higher marginal breakdown and increased susceptibility to wear following finishing procedures. Faced with the disadvantages brought by finishing and polishing procedures, it seems reasonable to try and find a technique to omit them. This paper presents clinical cases exhibiting a restorative technique that prevents, or at least reduces, the need for instrumentation of the restoration. In cases when finishing procedures are necessary, the restorations should be sealed with a specific resinous agent to impart new strength to the instrumented surface and improve marginal integrity. In addition, the suggested technique promotes a better occlusal anatomy than that attained with the traditional technique. PMID- 15130075 TI - Bonding to enamel and dentin using self-etching adhesive systems. AB - OBJECTIVES: This study investigated the effectiveness of three different dentin adhesive systems on the adhesion of resin composite to both dentin and enamel. METHOD AND MATERIALS: The flat dentin and enamel surfaces of 60 extracted human molar teeth were exposed by wet grinding with 600-grit silicon carbide paper. One total-etch self-priming adhesive system (Prime & Bond NT), one two-step self etching primer adhesive system (Clearfil SE Bond), and one "all-in-one" self etching adhesive system (Prompt L-Pop) were evaluated. Each bonding system was applied according to the manufacturer's instructions and followed by composite (TPH Spectrum) application. Twenty-four hours after bonding, the teeth were subjected to shear testing. There were 10 replicates for each group. RESULTS: Prompt L-Pop exhibited significantly higher bond strength values to enamel (27 +/ 4.2 MPa) than all other groups. There were no statistically significant differences for shear bond strength to dentin among adhesives. Prompt L-Pop showed the statistically significantly higher bond strength to enamel than dentin. There were no statistically significant differences between the enamel and dentin bond strengths of Clearfil SE Bond and Prime & Bond NT. CONCLUSIONS: The self-etching adhesive systems produced high bond strengths to human coronal dentin and ground enamel surfaces. These materials seem to be very promising for further clinical applications, and the results are very encouraging for the clinical success of these simplified adhesive systems. The self-etching adhesive systems produced even better bond strengths to both enamel and dentin than conventional total-etch systems, especially the "all-in-one" system, which produced the highest bond strength to enamel. PMID- 15130076 TI - Orthodontic extraction: conservative treatment of impacted mandibular third molar associated with a dentigerous cyst. A case report. AB - A 53-year-old man exhibited a large dentigerous cyst associated with a deep vertical impaction of his left mandibular third molar where a close anatomic relationship existed between the root and the mandibular canal. After consultation, it was decided to perform marsupialization of the cyst, to extrude the third molar orthodontically, and then to extract it. This combined orthodontic-surgical approach was successful. PMID- 15130077 TI - Accidental displacement of impacted maxillary and mandibular third molars. AB - Displacement of impacted third molars is frequently mentioned in oral and maxillofacial surgery textbooks, but rarely reported. However, should this complication arise in general practice, the clinician should not embark on potentially complicated and hazardous surgical procedures to retrieve the displaced tooth. Administration of prophylactic broad spectrum antibiotics and urgent referral to an oral and maxillofacial surgeon, is recommended. PMID- 15130078 TI - Acetic acid wash and chemiluminescent illumination as an adjunct to conventional oral soft tissue examination for the detection of dysplasia: a pilot study. AB - Population-based oral cancer screening appears to be a promising health promotion strategy (especially in high-risk individuals) with significant increases in quality-adjusted life years saved. However, the current protocol, conventional visual inspection, and palpation of oral soft tissues for the early detection of pre-malignant or malignant changes, appears to be deficient. The adjunctive application of technology to highlight such lesions may increase the diagnostic yield. The purpose of this pilot study was to collect data, which might support the hypothesis that oral soft tissues exhibit features similar to the cervical epithelium following an acetic acid wash and visual inspection under chemiluminescent illumination. The data provides strong evidence to support the hypothesis. Epithelium with hyperkeratinization, hyperparakeratinization, and/or chronic inflammatory infiltrate reflects the diffuse, low-level, blue-white chemiluminescent light more strongly and appears amplified. Similarly, epithelium with an altered nuclear-cytoplasmic ratio also reflects the diffuse, low-level, blue-white chemiluminescent light. In such cases, the lesions become clinically discernible and appear "acetowhite." Large-scale studies are required to further refine issues related to the selectivity and specificity of the technology. PMID- 15130079 TI - Glandular odontogenic cyst: case report and review of the literature. AB - Glandular odontogenic cyst (GOC) is a rare developmental cyst of the jaws. It is included in the World Health Organization (WHO) histologic typing of odontogenic tumors under the terms glandular odontogenic cyst or sialo-odontogenic cyst. The most common site of occurrence is the anterior mandible, and it occurs mostly in middle-aged people. A predilection for men is observed. Clinical findings are not specific, and an asymptomatic swelling is frequently observed. A unilocular or multilocular, well-defined radiolucency is usually seen. The microscopic features of GOC, particularly the morphology of the epithelium, strongly suggest an origin from the remains of dental lamina. GOC has an unpredictable and potentially aggressive nature, which may indicate a high tendency of recurrence. The treatment of choice is still controversial, varying from a curettage to local block excision. A long-term follow-up should be carried out. The aim of this article is to report a case of glandular odontogenic cyst that recurred four times and to emphasize the importance of long-term follow-up. The origin, epidemiology, clinical and radiographic aspects, and treatment of the GOC are also discussed. PMID- 15130080 TI - Saliva substitute in xerostomic patients with primary Sjogren's syndrome: a single-blind trial. AB - OBJECTIVE: To evaluate the efficacy of oral moisturizing gel (Oral Balance) in xerostomic patients with primary Sjogren's syndrome (SS). METHOD AND MATERIALS: Twenty-one xerostomic patients with primary SS were subjected to a single-blind trial in which the efficacy of Oral Balance gel in reducing xerostomia and xerostomia-related oral symptoms was compared with that of a placebo. Both gels were packaged identically and were indiscernible in appearance and taste. Xerostomia was confirmed for all the patients through measurement of stimulated whole saliva. Patients began using the Oral Balance gel three times a day for 90 days, and were then switched to a gel placebo to be used in the same way for the same length of time. Clinical response was evaluated through the patients' subjective assessment (improved, worsened, or unaltered) of both gels. RESULTS: Neither the Oral Balance gel nor the gel placebo affected the salivary output of the patients. The Oral Balance gel presented a substantial statistically significant advantage in the control of burning mouth, mastication, and swallowing. No statistically significant relief of the isolated sensation of oral dryness was established. CONCLUSION: Oral Balance is a useful tool in the management of dryness-related oral symptoms in primary SS, but there is room for enhancing the overall properties of topical preparations designed to reduce oral complaints in xerostomic patients. PMID- 15130081 TI - Removable prostheses for preschool children: report of two cases. AB - Dental prostheses are frequently used to avoid psychologic, speech, or swallowing problems in preschool children with considerable tooth loss. Two cases of preschool children are presented, involving multiple loss of primary teeth. The purpose of this study was to promote the correct development of the maxilla and mandible by using removable dental prostheses and to guide the eruption of the permanent molars. Removable acrylic prostheses were provided for two children, with a special metallic s-shaped handle (ansa), which guided the eruption of the first permanent molars. These prostheses were modified as the children grew. By replacing missing teeth, several oral functions were re-established, development of the maxilla and mandible was promoted, and each child could develop socially from a psychologic point of view. The use of removable dental prostheses in preschool-aged children presenting with considerable tooth loss can be a viable and successful treatment option. PMID- 15130082 TI - Preprosthetic therapy utilizing a temporary occlusal acrylic splint: a case report. AB - This case report describes the complex occlusal rehabilitation of a patient with signs and symptoms of temporomandibular disorders with utilization of an occlusal acrylic splint as a means of initial treatment for neuromuscular reprogramming and repositioning of the condyle within the mandibular fossa for occlusal stability, thus allowing adaptation to a new occlusal vertical dimension. PMID- 15130083 TI - Thirteen-year follow-up study of resin-bonded fixed partial dentures. AB - OBJECTIVE: The technique of resin-bonded fixed partial dentures (RBFPD) is a well accepted clinical technique to replace missing teeth. The survival rates reported in the literature vary widely, and the conclusions are sometimes conflicting. This study presents the clinical long-term performance of silicoated RBFPDs and also determines the main cause of failure. METHOD AND MATERIALS: Sixty-one patients with a total of 74 RBFPDs were either examined or requested to complete a questionnaire regarding their fixed partial dentures. Sixty-four were placed in the anterior region, and 10 in the posterior region. No more than one missing tooth in the posterior area and two missing teeth in the anterior region were replaced with RBFPDs. A retentive preparation was made on the abutment teeth. All the RBFPDs were adhesively seated. RESULTS: Eighteen RBFPDs failed after a mean observation time of 7.8 years (nine retention losses of one or more retainers, six carious lesions, and three veneer fractures occurred). Seven RBFPDs were rebonded, whereas the remaining 11 failures had to be replaced with conventional FPDs. Fifty-six RBFPDs were primary restorations, and seven secondary RBFPDs were used after the loss of the primary restoration. A mean survival rate better than 69% after a 13-year observation period was calculated. Including the rebonded restorations, a mean functional survival rate of 83% was estimated. A total of 18 failures (24.3%) of all restorations were observed, the main cause being loss of retention. CONCLUSION: Silicoated RBFPDs are a viable treatment means with an acceptable success expectancy. PMID- 15130084 TI - Dentin microhardness during and after whitening treatments. AB - OBJECTIVE: This in vitro study aimed to evaluate the effect of bleaching agents on dentin microhardness during and after bleaching. METHOD AND MATERIALS: Specimens were randomly assigned to seven groups (n = 15): Nite White Excel 2 Z [NW] 10% and 22%; Rembrandt [REM] 10% and 22%; Opalescence [OPA] 10% and 20%; and a placebo agent. The 42-day whitening treatment consisted of daily application of the agents to the dentin surfaces for 8 hours, followed by immersion in artificial saliva for 16 hours. After the bleaching treatment, specimens were kept immersed in artificial saliva for 14 days. Microhardness was measured at baseline, 8 hours, and 7, 14, 21, 28, 35, and 42 days of bleaching and during the posttreatment period (7 and 14 days). RESULTS: The analysis of variance for split plot showed a significant effect on the interaction between bleaching agent and time. Tukey's test and regression analyses revealed that during the bleaching period, the agents NW 10%, NW 22%, and OPA 20%, which did not differ from each other, did not alter dentin microhardness, showing constant microhardness values. There were no differences among REM 10%, REM 22%, and OPA 10%, which showed significant reductions in microhardness after day 14 compared to other agents. After bleaching procedures, there was an increase in dentin microhardness for all groups. CONCLUSION: Throughout the bleaching treatment, depending on the agent applied, dentin showed a transitory decrease in microhardness values. In the posttreatment period, artificial saliva presented a remineralizing effect on the bleached surfaces. PMID- 15130085 TI - Focal cemento-osseous dysplasia. PMID- 15130086 TI - The Drosophila melanogaster homologue of the human histo-blood group Pk gene encodes a glycolipid-modifying alpha1,4-N-acetylgalactosaminyltransferase. AB - Insects express arthro-series glycosphingolipids, which contain an alpha1,4 linked GalNAc residue. To determine the genetic basis for this linkage, we cloned a cDNA (CG17223) from Drosophila melanogaster encoding a protein with homology to mammalian alpha1,4-glycosyltransferases and expressed it in the yeast Pichia pastoris. Culture supernatants from the transformed yeast were found to display a novel UDP-GalNAc:GalNAcbeta1,4GlcNAcbeta1-R alpha-N acetylgalactosaminyltransferase activity when using either a glycolipid, p nitrophenylglycoside or an N-glycan carrying one or two terminal beta-N acetylgalactosamine residues. NMR and MS in combination with glycosidase digestion and methylation analysis indicate that the cloned cDNA encodes an alpha1,4-N-acetylgalactosaminyltransferase. We hypothesize that this enzyme and its orthologues in other insects are required for the biosynthesis of the N5a and subsequent members of the arthro-series of glycolipids as well as of N-glycan receptors for Bacillus thuringiensis crystal toxin Cry1Ac. PMID- 15130087 TI - MT1-MMP mediates MUC1 shedding independent of TACE/ADAM17. AB - MUC1, a transmembrane mucin, plays a critical role in embryo implantation, protection of mucosal epithelia from microbial and enzymic attack and various aspects of tumour progression. In some species, a decrease in uterine epithelial MUC1 protein and mRNA expression accompanies embryo implantation. In other species, such as rabbits and humans, MUC1 appears to be locally removed at blastocyst attachment sites, suggesting the action of a protease. We previously demonstrated that MUC1 is proteolytically released from the surface of a human uterine epithelial cell line, HES, and identified TACE/ADAM17 (where TACE stands for tumour necrosis factor-alpha converting enzyme and ADAM for A Disintegrin And Metalloprotease-like) as a constitutive and PMA-stimulated MUC1 sheddase [Thathiah, Blobel and Carson (2003) J. Biol. Chem. 274, 3386-3394]. Further characterization of the proteolytic activity(ies) mediating MUC1 release indicates that MUC1 shedding is also accelerated by the tyrosine phosphatase inhibitor pervanadate. Pervanadate, but not PMA, stimulates MUC1 shedding in TACE deficient cells, indicating activation of a metalloproteolytic activity(ies) distinct from TACE. Pervanadate-stimulated MUC1 release is inhibited by the TIMP 2 (tissue inhibitor of metalloprotease-2) and TIMP-3, but is unaffected by TIMP 1, consistent with the MT-MMPs (membrane-type matrix metalloproteases). Pervanadate stimulation of MUC1 shedding is absent from MUC1-transfected MT1-MMP deficient fibroblasts, but is restored after MUC1 and MT1-MMP co-transfection. Furthermore, overexpression of MT1-MMP in HES cells enhances pervanadate stimulated MUC1 release, and MT1-MMP co-localizes with MUC1 in vivo at the apical surface of receptive-phase human uterine epithelia. Taken together, these studies characterize a MUC1 sheddase activity in addition to TACE and identify MT1-MMP as a pervanadate-stimulated MUC1 sheddase. PMID- 15130088 TI - Functional analysis of Chinese hamster phosphatidylserine synthase 1 through systematic alanine mutagenesis. AB - PtdSer (phosphatidylserine) synthesis in mammalian cells occurs through the exchange of L-serine with the base moieties of phosphatidylcholine and phosphatidylethanolamine, which is catalysed by PSS (PtdSer synthase) 1 and 2 respectively. PtdSer synthesis in intact cells and an isolated membrane fraction was inhibited by exogenous PtdSer, indicating that feedback control is involved in the regulation of PtdSer biosynthesis. PSS 1 and 2 are similar in amino acid sequence, with an identity of 32%; however, due to a lack of homology with other known enzymes, their amino acid sequences do not provide information on their catalytic and regulatory mechanisms. In the present study, to identify amino acid residues crucial for the activity and/or regulation of PSS 1, we systematically introduced mutations into a Chinese hamster PSS 1 cDNA clone; namely, each of the 66 polar amino acid residues common to PSS 2 was replaced with an alanine residue. On analysis of Chinese hamster ovary cells transfected with each of the alanine mutant clones, we identified eight amino acid residues (His-172, Glu-197, Glu-200, Asn-209, Glu-212, Asp-216, Asp-221 and Asn-226) as those crucial for the enzyme reaction or the maintenance of the correct structure required for serine base-exchange activity. Among these residues, Asn-209 was suggested to be involved in the recognition and/or binding of free L-serine. We also identified six amino acid residues (Arg-95, His-97, Cys-189, Arg-262, Gln-266 and Arg-336) as those important for regulation of PSS 1. In addition, we found that the alanine mutations at Tyr-111, Asp-166, Arg-184, Arg-323, and Glu-364 affected the production and/or stability of PSS 1 in Chinese hamster ovary cells. PMID- 15130089 TI - Calmidazolium and arachidonate activate a calcium entry pathway that is distinct from store-operated calcium influx in HeLa cells. AB - Agonists that deplete intracellular Ca2+ stores also activate Ca2+ entry, although the mechanism by which store release and Ca2+ influx are linked is unclear. A potential mechanism involves 'store-operated channels' that respond to depletion of the intracellular Ca2+ pool. Although SOCE (store-operated Ca2+ entry) has been considered to be the principal route for Ca2+ entry during hormonal stimulation of non-electrically excitable cells, recent evidence has suggested that alternative pathways activated by metabolites such as arachidonic acid are responsible for physiological Ca2+ influx. It is not clear whether such messenger-activated pathways exist in all cells, whether they are truly distinct from SOCE and which metabolites are involved. In the present study, we demonstrate that HeLa cells express two pharmacologically and mechanistically distinct Ca2+ entry pathways. One is the ubiquitous SOCE route and the other is an arachidonate-sensitive non-SOCE. We show that both these Ca2+ entry pathways can provide long-lasting Ca2+ elevations, but that the channels are not the same, based on their differential sensitivity to 2-aminoethoxydiphenyl borate, LOE-908 [(R,S)-(3,4-dihydro-6,7-dimethoxy-isochinolin-1-yl)-2-phenyl-N,N-di[2-(2,3,4 trimethoxyphenyl)ethyl]acetamid mesylate] and gadolinium. In addition, non-SOCE and not SOCE was permeable to strontium. Furthermore, unlike SOCE, the non-SOCE pathway did not require store depletion and was not sensitive to displacement of the endoplasmic reticulum from the plasma membrane using jasplakinolide or ionomycin pretreatment. These pathways did not conduct Ca2+ simultaneously due to the dominant effect of arachidonate, which rapidly curtails SOCE and promotes Ca2+ influx via non-SOCE. Although non-SOCE could be activated by exogenous application of arachidonate, the most robust method for stimulation of this pathway was application of the widely used calmodulin antagonist calmidazolium, due to its ability to activate phospholipase A2. PMID- 15130090 TI - Recruitment of the cross-linked opsonic receptor CD32A (FcgammaRIIA) to high density detergent-resistant membrane domains in human neutrophils. AB - We have previously shown that CD32A (or FcgammaRIIA), one of the main opsonin receptors, was rapidly insolubilized and degraded in intact neutrophils after its cross-linking. In view of these experimental difficulties, the early signalling steps in response to CD32A activation were studied in purified plasma membranes of neutrophils. After CD32A cross-linking in these fractions, the tyrosine phosphorylation of two major substrates, the receptor itself and the tyrosine kinase Syk, was observed. Phosphorylation of these two proteins was observed only in the presence of orthovanadate, indicating the presence, in the membranes, of one or more tyrosine phosphatases that maintain CD32A dephosphorylation. The tyrosine phosphorylation of these two proteins was inhibited by the Src kinase inhibitor, 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (PP2). The ligation of CD32A led to its recruitment to a previously uncharacterized subset of high-density flotillin-1-positive DRMs (detergent-resistant membranes). The changes in the solubility properties of CD32A were observed in the absence of added ATP; therefore, they were probably not secondary to the tyrosine phosphorylation of the receptor, rather they preceded it. Src kinases as well as Syk were constitutively present in DRMs of high and low density and no evident changes in their distribution were detected after cross-linking of CD32A. Pretreatment of plasma membranes with methyl-beta-cyclodextrin did not inhibit the recruitment of CD32A to DRMs, although it led to the loss of the Src kinase Lyn from these fractions. In addition, methyl-beta-cyclodextrin inhibited the tyrosine phosphorylation of CD32A and Syk induced by cross-linking of CD32A. This membrane model allowed us to observe a movement of CD32A from detergent-soluble regions of the membranes to DRMs, where it joined Src kinases and Syk and became tyrosine-phosphorylated. PMID- 15130091 TI - Identification of the actin-binding domain of Ins(1,4,5)P3 3-kinase isoform B (IP3K-B). AB - Dewaste et al. [Dewaste, Moreau, De Smedt, Bex, De Smedt, Wuytaack, Missiaen and Erneux (2003) Biochem. J. 374, 41-49] showed that over-expressed EGFP (enhanced green fluorescent protein) fused to Ins(1,4,5)P3 3-kinase B (IP3K-B) co-localizes with the cytoskeleton, as well as with the endoplasmic reticulum and the plasma membrane. The domains responsible for these subcellular localizations are not yet identified. For the endogenous enzyme, we confirmed both actin and endoplasmic reticulum localization by employing a high affinity antibody against IP3K-B. F actin targeting is exclusively dependent on the non-catalytic N-terminal region of IP3K-B. By expressing fragments of this N-terminal domain as EGFP-fusion proteins and inspecting transfected cells by confocal microscopy, we characterized a distinct 63-amino-acid domain comprising amino acids 108-170 of the enzyme which is responsible for F-actin targeting. A truncation of this fragment from both sides revealed that the full size of this segment is essential for this function. Deletion of this segment in a full-length over-expressed IP3K B-EGFP-fusion protein completely abolished F-actin interaction. Direct interaction of this actin-binding segment with only F-actin, but not with G actin, was observed in vitro using a bacterially expressed, affinity-purified GST (glutathione S-transferase)-Rattus norvegicus IP3K (aa 108-170) fusion protein. Helix-breaking mutations within this isolated segment abolished the F-actin binding properties both in vitro and when over-expressed in cells, indicating that an intact secondary structure is essential for actin targeting. The segment shows sequence similarities to the actin-binding region in IP3K-A, but no similarity to other actin-binding domains. PMID- 15130092 TI - Functional analysis of peroxisome-proliferator-responsive element motifs in genes of fatty acid-binding proteins. AB - Retinoic acids and long-chain fatty acids are lipophilic agonists of nuclear receptors such as RXRs (retinoic X receptors) and PPARs (peroxisome-proliferator activated receptors) respectively. These agonists are also ligands of intracellular lipid-binding proteins, which include FABPs (fatty acid-binding proteins). We reported previously that L (liver-type)-FABP targets fatty acids to the nucleus of hepatocytes and affects PPARalpha activation, which binds together with an RXR subtype to a PPRE (peroxisome-proliferator-responsive element). In the present study, we first determined the optimal combination of murine PPAR/RXR subtypes for binding to known murine FABP-PPREs and to those found by computer search and then tested their in vitro functionality. We show that all PPARs bind to L-FABP-PPRE, PPARalpha, PPARgamma1 and PPARgamma2 to A (adipocyte-type)-FABP PPRE. All PPAR/RXR heterodimers transactivate L-FABP-PPRE, best are combinations of PPARalpha with RXRalpha or RXRgamma. In contrast, PPARalpha heterodimers do not transactivate A-FABP-PPRE, best combinations are of PPARgamma1 with RXRalpha and RXRgamma, and of PPARgamma2 with all RXR subtypes. We found that the predicted E (epidermal-type)- and H (heart-type)-FABP-PPREs are not activated by any PPAR/RXR combination without or with the PPAR pan-agonist bezafibrate. In the same way, C2C12 myoblasts transfected with promoter fragments of E-FABP and H FABP genes containing putative PPREs are also not activated through stimulation of PPARs with bezafibrate applied to the cells. These results demonstrate that only PPREs of L- and A-FABP promoters are functional, and that binding of PPAR/RXR heterodimers to a PPRE in vitro does not necessarily predict transactivation. PMID- 15130093 TI - Medication management by graduate nurses: before, during and following medication administration. AB - The purpose of the present study was to explore graduate nurses' perceptions of their medication management activities in the acute care context. A qualitative research design with a semistructured interview schedule was used to elicit information from participants. The sampling population consisted of graduate nurses involved in direct patient care in medical and surgical wards of a Melbourne metropolitan teaching hospital, completing a graduate nurse program. Twelve graduate nurses participated in the interviews. Two major themes emerged: (i). monitoring medications and (ii). interventions for patient care. The findings indicate that graduate nurses are required to address several facets of the medication management role in their daily practice. It is pertinent to examine ward dynamics to ensure that graduate nurses have ready access to experienced health care professionals. Through collegial support, graduate nurses should also be encouraged to critically examine the different possibilities when making clinical judgments about monitoring patient medications. PMID- 15130094 TI - Parents' experiences of asthma: process from chaos to coping. AB - The aim of the present qualitative study was to describe nine parents' everyday experiences of living with a child suffering from asthma. Data were collected by means of in-depth interviews and phenomenological content analysis. Four main themes emerged: feelings of uncertainty, helplessness and guilt; the need for support and help from healthcare professionals; adaptation to everyday life; and the development of coping strategies. In addition, two subthemes; trying out and seeking information, emerged. Trying out was found to be an important strategy for parents in managing the illness. In encounters with healthcare professionals, parents felt that they were not respected and that their competence was questioned. In conclusion, this study emphasizes the importance of a mutual dialogue between healthcare professionals and parents to enable the parents to develop the competence necessary to care for their child. PMID- 15130095 TI - Together we are heard: effectiveness of daily 'language' groups in a community preschool. AB - Strong oral language skills are a prerequisite for successful literacy and there is a strong interdependence between oral language acquisition and emergent literacy development. Ramifications of this are that children with language impairments are at great risk for difficulties in learning to read and write, with problems often persisting throughout the school years into adulthood. The Together we are heard program involved improving each child's oral language skills through group sessions facilitated by a speech pathologist on a daily basis at preschool. The aim of the present research was to determine the effectiveness of the program to identify the best way to assist children to develop appropriate language skills. The study showed that the children improved significantly in all four levels of the Preschool Language Assessment Inventory (PLAI). Importantly, the program was effective for both genders and there was no difference in the success of Indigenous children when compared to their European counterparts. There is a strong recommendation for further research and to expand such programs, particularly in areas that target children from impoverished and deprived environmental backgrounds. PMID- 15130096 TI - Hand dermatitis among female nursing students in tropical Australia. AB - Although hand dermatitis is known to affect nursing students worldwide, the prevalence among their Australian counterparts has not been elucidated and the relative contribution of tropical environments is unclear. Therefore, we conducted the first investigation of hand dermatitis among 232 female, undergraduate nurses in tropical Australia using a previously validated methodology. Hand dermatitis prevalence rose from 10.8% in the first year to 27.4% by the third year and averaged 18.5% across all three grades. Pre-existing atopic dermatitis caused a 7.8-fold risk increase during logistic regression analysis (odds ratio [OR] 7.8, 95% confidence interval [CI] 3.0-21.1, P < 0.0001). Overall, this study suggests that hand dermatitis is less common among tropical Australian nursing students than their counterparts around the world. The identification of atopic dermatitis as a hand dermatitis risk factor was, however, consistent with previous research conducted in non-tropical areas. PMID- 15130097 TI - Assessments and nursing care for right brain-damaged stroke patients: focusing on neglect and related symptoms. AB - The purpose of present study was to identify the behavioural characteristics and examine assessments of nursing care among right brain-damaged stroke patients with neglect within 1 year following initial onset. Characteristics related to higher cerebral dysfunction were extracted and patient grouping was attempted through cognition (Mini-Mental State Examination) and physical function (Barthel Index) plots. The characteristics and implications for nursing care for each group were analyzed. Twelve patients were classified into four groups using cognitive-physical function evaluations together with subjective and objective data associated with neglect. 'Neglect' has been regarded as a disorder of spatial perception. However, the characteristics observed in Group 2 suggest that 'neglect' may have another facet, manifesting as disorders in perceiving continuous spatio-temporal changes of an action and comprehending the context of a situation. In these patients, although 'by calling attention' is the conventional care, more appropriate care should be provided based on an assessment of cognitive-physical function and spatio-temporal recognition of an action. PMID- 15130098 TI - Living with Huntington's disease: need for supportive care. AB - Huntington's disease is a genetic, neurological disorder characterized by mid life onset, involuntary movements, cognitive decline, behavioral disturbance, and inexorable progression. The impact of Huntington's disease is devastating for individuals and their families as it is a disease with a long trajectory; many young people are aware that they may develop the illness for years before there are obvious symptoms. There is therefore ample opportunity to plan and choreograph the care and supportive services for people with Huntington's disease and their families. The present study was conducted to explore the needs for palliative (supportive) care service provision of people with Huntington's disease and their families/informal carers. Six people with the disease, 19 informal carers and seven health care workers with specialized knowledge took part in individual, semistructured interviews, which were analyzed thematically. Themes were: (i). adjusting to the impact of the illness; (ii). surviving the search for essential information; (iii). gathering practical support from many sources; (iv). bolstering the spirit; (v). choreographing individual care and; (vi). fearing the future. Our findings demonstrate that palliative care services for people with Huntington's disease and their informal carers need to provide expert psychological and practical support and perhaps most importantly, be flexible, adequately planned and choreographed. PMID- 15130099 TI - Ethical concerns: comparison of values from two cultures. AB - The present study was a secondary analysis of data from two phenomenological studies of nurses in the USA and Japan. The study incorporated hermeneutics and feminist methodologies to answer the following questions. Are there common values and ethical concerns and values within the nursing cultures of Japan and the USA? What are some commonalities and differences between Japanese nurses' ethical concerns and those of American nurses? Findings indicated that nurses from the USA and Japan share common values and ethical concerns as professional nurses, including competence, respect for the patient as a person, responsibility, relationship and connection, importance of the family, caring, good death, comfort, truth-telling, understanding the patient/situation, and anticipatory care. Although ethical concerns are similar, related background meanings and actions often look different between cultures; truth-telling is described as an example. Nurses in each country also hold unique values not found in the nursing practice of the other country. Understanding these commonalities and differences is critical for the development of global nursing ethics. PMID- 15130100 TI - Impact of in-depth interviews on the interviewer: roller coaster ride. AB - The authors investigated the experiences of parents with children/adult children in metropolitan Sydney, Australia who were living with, or had recovered from, an eating disorder. During regular team meetings, the research assistant who conducted the interviews had described her reactions which led the research team to investigate her experience in more depth. The aim of the present paper was to explore the impact on the research assistant who conducted 22 in-depth interviews with the parents. One of the members of the research team interviewed the research assistant to elicit her reactions. The interview was content analyzed and the following themes were identified: (i). appreciation of an egalitarian model of research; (ii). the emotions expressed by the research assistant; (iii). making sense of the inexplicable and (iv). reflections and comparison to her own life role. The research team would like to advance the theory that the adoption of a formal debriefing mechanism be integrated into the qualitative research process. PMID- 15130101 TI - Health care seeking behavior of Korean women with lymphedema. AB - The present biocultural study aimed to describe the health care use patterns of women with lymphedema. Data came from interviews and participant observations with eight key informants between February 2000 and February 2002. Analyzing the process of seeking health care, this paper explored how Korean women with lymphedema make use of all the available resources in the three sectors of the health care system: professional, folk and popular health. In these three sectors of the health care system, informants showed different patterns of behavior. In the professional health care sector, they behave based on scientific Western medicine and holistic herbal medical frameworks. Informants want scientific technological treatment from a Westernized doctor and perfect humanistic and holistic treatment from a herbal doctor. In the folk sector, informants' behavior is ruled by a pragmatic and supernatural framework. Informants seek religious healers who have strong spirituality and non-religious healers who have experience and skills. Informants complied with these healer's remedies based on efficacy and empirical healing evidence. In the popular sector of the health care system, informants behave based on their concept of illness and rules of daily life. They believe lymphedema comes from poor blood circulation and they want to be regarded as members of society, not as patients with lymphedema. Therefore, informants practised popular remedies that they believed were good for promoting blood circulation and keeping their social network active. This description about health care seeking behaviors being embedded in Korean socio-medical culture can serve to understand patients with other chronic health problems. With these results, we can put a bridge over the river of cultural conflict between health professionals and patients. PMID- 15130114 TI - Environmental signals and regulatory pathways that influence biofilm formation. AB - In nature, bacteria often exist as biofilms. Here, we discuss the environmental signals and regulatory proteins that affect both the initiation of bacterial biofilm formation and the nature of the mature biofilm structure. Current research suggests that the environmental signals regulating whether bacterial cells will initiate a biofilm differ from one bacterial species to another. This may allow each bacterial species to colonize its preferred environment efficiently. In contrast, some of the environmental signals that have currently been identified to regulate the structure of a mature biofilm are nutrient availability and quorum sensing, and are not species specific. These environmental signals evoke changes in the nature of the mature biofilm that may ensure optimal nutrient acquisition. Nutrient availability regulates the depth of the biofilm in such a way that the maximal number of cells in a biofilm appears to occur at suboptimal nutrient concentrations. At either extreme, nutrient-rich or very nutrient-poor conditions, greater numbers of cells are in the planktonic phase where they have greater access to the local nutrients or can be distributed to a new environment. Similarly, quorum-sensing control of the formation of channels and pillar-like structures may ensure efficient nutrient delivery to cells in a biofilm. PMID- 15130115 TI - Untangling intracellular DNA topology. AB - The biochemical steps by which bacterial topoisomerases alter the topology of DNA are well known. However, it has been a more vexing task to establish physiological roles and sites of action of the different topoisomerases within the context of the bacterial cell cycle. This difficulty can be attributed in part to the redundancy among the activities of the different enzymes. In this microreview, we will focus on recent progress in understanding the topological structure of the chromosome, analysis of topoisomerase mechanism in single molecule assays and recent data on the regulation and integration of topoisomerase activity within the cell cycle that have all brought a new perspective to the action of topoisomerases in the bacterial cell. PMID- 15130116 TI - Cell-to-cell signalling in Escherichia coli and Salmonella enterica. AB - Cell-to-cell signalling in prokaryotes that leads to co-ordinated behaviour has been termed quorum sensing. This type of signalling can have profound impacts on microbial community structure and host-microbe interactions. The Gram-negative quorum-sensing systems were first discovered and extensively characterized in the marine Vibrios. Some components of the Vibrio systems are present in the classical genetic model organisms Escherichia coli and Salmonella enterica. Both organisms encode a signal receptor of the LuxR family, SdiA, but not a corresponding signal-generating enzyme. Instead, SdiA of Salmonella detects and responds to signals generated only by other microbial species. Conversely, E. coli and Salmonella encode the signal-generating component of a second system (a LuxS homologue that generates AI-2), but the sensory apparatus for AI-2 differs substantially from the Vibrio system. The only genes currently known to be regulated by AI-2 in Salmonella encode an active uptake and modification system for AI-2. Therefore, it is not yet clear whether Salmonella uses AI-2 as a signal molecule or whether AI-2 has some other function. In E. coli, the functions of both SdiA and AI-2 are unclear due to pleiotropy. Genetic strategies to identify novel signalling systems have been performed with E. coli and Providencia stuartii. Several putative signalling systems have been identified, one that uses indole as a signal and another that releases what appears to be a peptide. The latter system has homologues in E. coli and Salmonella, as well as other bacteria, plants and animals. In fact, the protease components from Providencia and Drosophila are functionally interchangeable. PMID- 15130117 TI - Genome-wide analysis of transcriptional hierarchy and feedback regulation in the flagellar system of Helicobacter pylori. AB - The flagellar system of Helicobacter pylori, which comprises more than 40 mostly unclustered genes, is essential for colonization of the human stomach mucosa. In order to elucidate the complex transcriptional circuitry of flagellar biosynthesis in H. pylori and its link to other cell functions, mutants in regulatory genes governing flagellar biosynthesis (rpoN, flgR, flhA, flhF, HP0244) and whole-genome microarray technology were used in this study. The regulon controlled by RpoN, its activator FlgR (FleR) and the cognate histidine kinase HP0244 (FleS) was characterized on a genome-wide scale for the first time. Seven novel genes (HP1076, HP1233, HP1154/1155, HP0366/367, HP0869) were identified as belonging to RpoN-associated flagellar regulons. The hydrogenase accessory gene HP0869 was the only annotated non-flagellar gene in the RpoN regulon. Flagellar basal body components FlhA and FlhF were characterized as functional equivalents to master regulators in H. pylori, as their absence led to a general reduction of transcripts in the RpoN (class 2) and FliA (class 3) regulons, and of 24 genes newly attributed to intermediate regulons, under the control of two or more promoters. FlhA- and FlhF-dependent regulons comprised flagellar and non-flagellar genes. Transcriptome analysis revealed that negative feedback regulation of the FliA regulon was dependent on the antisigma factor FlgM. FlgM was also involved in FlhA- but not FlhF-dependent feedback control of the RpoN regulon. In contrast to other bacteria, chemotaxis and flagellar motor genes were not controlled by FliA or RpoN. A true master regulator of flagellar biosynthesis is absent in H. pylori, consistent with the essential role of flagellar motility and chemotaxis for this organism. PMID- 15130118 TI - Differential recognition of members of the carcinoembryonic antigen family by Afa/Dr adhesins of diffusely adhering Escherichia coli (Afa/Dr DAEC). AB - Little is known about the molecular bases underlying the virulence of diffusely adhering Escherichia coli (DAEC) harbouring the Afa/Dr family of adhesins. These adhesins recognize as receptors the GPI-anchored proteins CD55 (decay accelerating factor, DAF) and CD66e (carcinoembryonic antigen, CEA). CD66e is a member of the CEA-related cell adhesion molecules (CEACAM) family, comprising seven members. We analysed the interactions of Afa/Dr DAEC with the CEACAMs using CEACAM-expressing CHO and HeLa cells. The results demonstrate that only E. coli expressing a subfamily of Afa/Dr adhesins, named here Afa/Dr-I, including Dr, F1845 and AfaE-III adhesins, bound onto CHO cells expressing CEACAM1, CEA or CEACAM6. Whereas all the Afa/Dr adhesins elicit recruitment of CD55 around adhering bacteria, only the Afa/Dr-I subfamily elicits the recruitment of CEACAM1, CEA and CEACAM6. In addition, although CEACAM3 is not recognized as a receptor by the subfamily of Afa/Dr adhesins, it is recruited around bacteria in HeLa cells. The recruited CEACAM1, CEA and CEACAM6 around adhering bacteria resist totally or in part a detergent extraction, whereas the recruited CEACAM3 does not. Finally, the results show that recognition of CEA and CEACAM6, but not CEACAM1, is accompanied by tight attachment to bacteria of cell surface microvilli-like extensions, which are elongated. Moreover, recognition of CEA is accompanied by an activation of the Rho GTPase Cdc42 and by a phosphorylation of ERM, which in turn elicit the observed cell surface microvilli-like extensions. PMID- 15130119 TI - Dissecting the loci of low-level quinine resistance in malaria parasites. AB - Quinine (QN) remains effective against Plasmodium falciparum, but its decreasing efficacy is documented from different continents. Multiple genes are likely to contribute to the evolution of QN resistance. To locate genes contributing to QN response variation, we have searched a P. falciparum genetic cross for quantitative trait loci (QTL). Results identify additive QTL in segments of chromosomes (Chrs) 13, 7 and 5, and pairwise effects from two additional loci of Chrs 9 and 6 that interact, respectively, with the QTL of Chrs 13 and 7. The mapped segments of Chrs 7 and 5 contain pfcrt, the determinant of chloroquine resistance (CQR), and pfmdr1, a gene known to affect QN responses. Association of pfcrt with a QTL of QN resistance supports anecdotal evidence for an evolutionary relationship between CQR and reduced QN sensitivity. The Chr 13 segment contains several candidate genes, one of which (pfnhe-1) encodes a putative Na(+)/H(+) exchanger. A repeat polymorphism in pfnhe-1 shows significant association with low QN response in a collection of P. falciparum strains from Asia, Africa and Central and South America. Dissection of the genes and modifiers involved in QN response will require experimental strategies that can evaluate multiple genes from different chromosomes in combination. PMID- 15130120 TI - IS6110 functions as a mobile, monocyte-activated promoter in Mycobacterium tuberculosis. AB - The mobile insertion sequence, IS6110, is an important marker in tracking of Mycobacterium tuberculosis strains. Here, we demonstrate that IS6110 can upregulate downstream genes through an outward-directed promoter in its 3' end, thus adding to the significance of this element. Promoter activity was orientation dependent and was localized within a 110 bp fragment adjacent to the right terminal inverted repeat. Transcripts from this promoter, named OP6110, begin approximately 85 bp upstream of the 3' end of IS6110. Use of green fluorescent protein (GFP) expression constructs showed that OP6110 was upregulated in M. tuberculosis during growth in human monocytes and in late growth phases in broth. Analysis of natural insertion sites in M. tuberculosis showed that IS6110 upregulated expression of several downstream genes during growth in human monocytes, including Rv2280 in H37Rv and the PE-PGRS gene, Rv1468c, in the clinical strain 210, which is a member of the Beijing family. Transcription between IS6110 and downstream genes was confirmed by reverse transcription polymerase chain reaction. The ability to activate genes during infection suggests that IS6110 has the potential to influence growth characteristics of different strains, and indicates another mechanism by which IS6110 can impact M. tuberculosis evolution. PMID- 15130121 TI - Phage T4 early promoters are resistant to inhibition by the anti-sigma factor AsiA. AB - Phage T4 early promoters are transcribed in vivo and in vitro by the Escherichia coli RNA polymerase holoenzyme Esigma(70). We studied in vitro the effects of the T4 anti-sigma(70) factor AsiA on the activity of several T4 early promoters. In single-round transcription, promoters motB, denV, mrh.2, motA wild type and UP element-deleted motA are strongly resistant to inhibition by AsiA. The alpha-C terminal domain of Esigma(70) is crucial to this resistance. DNase I footprinting of Esigma(70) and Esigma(70)AsiA on motA and mrh.2 shows extended contacts between the holoenzyme with or without AsiA and upstream regions of these promoters. A TG --> TC mutation of the extended -10 motif in the motA UP element deleted promoter strongly increases susceptibility to inhibition by AsiA, but has no effect on the motA wild-type promoter: either the UP element or the extended 10 site confers resistance to AsiA. Potassium permanganate reactivity shows that the two structure elements are not equivalent: with AsiA, the motA UP element deleted promoter opens more slowly whereas the motA TC promoter opens like the wild type. Changes in UV laser photoreactivity at position +4 on variants of motA reveal an analogous distinction in the roles of the extended -10 and UP promoter elements. PMID- 15130122 TI - Proteomic screening and identification of differentially distributed membrane proteins in Escherichia coli. AB - Bacteria show asymmetric subcellular distribution of many proteins involved in diverse cellular processes such as chemotaxis, motility, actin polymerization, chromosome partitioning and cell division. In many cases, the specific subcellular localization of these proteins is critical for proper regulation and function. Although cellular organization of the bacterial cell clearly plays an important role in cell physiology, systematic studies to uncover asymmetrically distributed proteins have not been reported previously. In this study, we undertook a proteomics approach to uncover polar membrane proteins in Escherichia coli. We identified membrane proteins enriched in E. coli minicells using a combination of two-dimensional electrophoresis and mass spectrometry. Among a total of 173 membrane protein spots that were consistently detected, 36 spots were enriched in minicell membranes, whereas 15 spots were more abundant in rod cell membranes. The minicell-enriched proteins included the inner membrane proteins MCPs, AtpA, AtpB, YiaF and AcrA, the membrane-associated FtsZ protein and the outer membrane proteins YbhC, OmpW, Tsx, Pal, FadL, OmpT and BtuB. We immunolocalized two of the minicell-enriched proteins, OmpW and YiaF, and showed that OmpW is a bona fide polar protein whereas YiaF displays a patchy membrane distribution with a polar and septal bias. PMID- 15130124 TI - The effect of host-encoded nucleoid proteins on transposition: H-NS influences targeting of both IS903 and Tn10. AB - Nucleoid proteins are small, abundant, DNA-binding proteins that profoundly affect the local and global structure of the chromosome, and play a major role in gene regulation. Although several of these proteins have been shown to enhance assembly of transpososomes before initiating transposition, no systematic survey has been carried out examining the in vivo role(s) of these proteins in transposition. We have examined the requirement of the six most abundant nucleoid proteins in transposition for three different transposons, IS903, Tn10 and Tn552. Most notably, H-NS was required for efficient transposition of all three elements in a papillation assay, suggesting a general role for H-NS in bacterial transposition. Further studies indicated that H-NS was exerting its effect on target capture. Targeting preferences for IS903 into the Escherichia coli chromosome were dramatically altered in the absence of H-NS. In addition, the alterations observed in the IS903 target profile emphasized the important role that H-NS plays in chromosome organization. A defect in target capture was also inferred for Tn10, as an excised transposon fragment, a precursor to target capture, accumulated in in vivo induction assays. Furthermore, a transposase mutant that is known to increase target DNA bending and to relax target specificity eliminated this block to target capture. Together, these results imply a role for H-NS in target capture, either by providing regions of DNA more accessible to transposition or by stabilizing transpososome binding to captured targets immediately before strand transfer. PMID- 15130123 TI - Branching sites and morphological abnormalities behave as ectopic poles in shape defective Escherichia coli. AB - Certain mutants in Escherichia coli lacking multiple penicillin-binding proteins (PBPs) produce misshapen cells containing kinks, bends and branches. These deformed regions exhibit two structural characteristics of normal cell poles: the peptidoglycan is inert to dilution by new synthesis or turnover, and a similarly stable patch of outer membrane caps the sites. To test the premise that these aberrant sites represent biochemically functional but misplaced cell poles, we assessed the intracellular distribution of proteins that localize specifically to bacterial poles. Green fluorescent protein (GFP) hybrids containing polar localization sequences from the Shigella flexneri IcsA protein or from the Vibrio cholerae EpsM protein formed foci at the poles of wild-type E. coli and at the poles and morphological abnormalities in PBP mutants. In addition, secreted wild type IcsA localized to the outer membrane overlying these aberrant domains. We conclude that the morphologically deformed sites in these mutants represent fully functional poles or pole fragments. The results suggest that prokaryotic morphology is driven, at least in part, by the controlled placement of polar material, and that one or more of the low-molecular-weight PBPs participate in this process. Such mutants may help to unravel how particular proteins are targeted to bacterial poles, thereby creating important biochemical and functional asymmetries. PMID- 15130125 TI - Structural tolerance of bacterial autotransporters for folded passenger protein domains. AB - In this report we investigate the capacity of bacterial autotransporters (AT) to translocate folded protein domains across the outer membrane (OM). Polypeptides belonging to the AT family contain a C-terminal domain that supports the secretion of the N-domain (the passenger) across the OM of Gram-negative bacteria. Despite some controversial data, it has been widely accepted that N passenger domains of AT must be unfolded and devoid of disulphide bonds for efficient translocation. To address whether or not AT are able to translocate folded protein domains across the OM, we employed several types of recombinant antibodies as heterologous N-passengers of the transporter C-domain of IgA protease (C-IgAP) of Neisseria gonorroheae. The N-domains used were single chain Fv fragments (scFv) and variable mono-domains derived from camel antibodies (V(HH)) selected on the basis of their distinct and defined folding properties (i.e. enhanced solubility, stability and presence or not of disulphide bonds). Expression of these hybrids in Escherichia coli shows that stable scFv and V(HH) domains are efficiently (>99%) translocated towards the bacterial surface regardless of the presence or not of disulphide bonds on their structure. Antigen binding assays demonstrate that surface-exposed scFv and V(HH) domains are correctly folded and thus able to bind their cognate antigens. Expression of scFv or V(HH)-C-IgAP hybrids in E. coli dsbA or fkpA mutant cells reveals that these periplasmic protein chaperones fold these N-domains before their translocation across the OM. Furthermore, large N-passengers composed of strings of V(HH) domains were secreted in a folded state by AT with no loss of efficacy (>99%) despite having multiple disulphide bonds. Thus AT can efficiently translocate toward the cell surface folded N-passengers composed of one, two or three immunoglobulin (Ig) domains, each with a folded diameter between approximately 2 nm and having disulphide bonds. This tolerance for folded protein domains of approximately 2 nm fits with the diameter of the central hydrophilic channel proposed for the ring-like oligomeric complex assembled by C-IgAP in the OM. PMID- 15130126 TI - Fur functions as an activator and as a repressor of putative virulence genes in Neisseria meningitidis. AB - Fur is a well-known iron-responsive repressor of gene transcription, which is used by many bacteria to respond to the low-iron environment that pathogens encounter during infection. Four promoters of Neisseria meningitidis predicted to have Fur-binding boxes were selected to study the molecular interactions between Fur and the promoter regions of genes expected to play a central role in survival and pathogenesis. We demonstrate that Fur acts not only as a repressor, but also as an activator of gene expression both in vivo and in vitro. We report that Fur binds to operators located upstream of three promoters that are positively regulated in vivo by Fur and iron, whereas Fur binds to an operator overlapping the classically iron-repressed tbp promoter. Deletion of the upstream operator in the norB promoter abolished activation of transcription in vivo in response to iron and in vitro in response to Fur. The role of such a dual mechanism of Fur regulation during infection is discussed. PMID- 15130127 TI - The Bacillus subtilis sigmaW anti-sigma factor RsiW is degraded by intramembrane proteolysis through YluC. AB - The Bacillus subtilis sigma(W) regulon is induced by different stresses such as alkaline shock, salt shock, phage infection and certain antibiotics that affect cell wall biosynthesis. The activity of the alternative, extracytoplasmic function (ECF) sigma factor sigma(W) is modulated by a specific anti-sigma factor (RsiW or YbbM) encoded by the rsiW (ybbM) gene located immediately downstream of sigW. The RsiW membrane topology was determined, and a specific reporter system for RsiW function was constructed. Experiments using the yeast two-hybrid system suggested a direct interaction of sigma(W) with the cytoplasmic part of RsiW. Analysis of truncated forms of the RsiW protein revealed that sigma(W) induction by alkaline shock is dependent on both the transmembrane and the extracytoplasmic domain of RsiW. Western blot and pulse-chase experiments demonstrated degradation of RsiW after an alkaline shock. A B. subtilis mutant strain deleted for the Escherichia coli yaeL orthologue yluC, encoding a transmembrane protease, was defective in inducing a sigma(W)-controlled promoter after alkaline shock and accumulated a membrane-bound truncated form of RsiW, suggesting that the activity of sigma(W) is controlled by the proteolysis of RsiW by at least two different proteolytic steps. PMID- 15130128 TI - Characterization of an inducible vancomycin resistance system in Streptomyces coelicolor reveals a novel gene (vanK) required for drug resistance. AB - Vancomycin is the front-line therapy for treating problematic infections caused by methicillin-resistant Staphylococcus aureus (MRSA), and the spread of vancomycin resistance is an acute problem. Vancomycin blocks cross-linking between peptidoglycan intermediates by binding to the D-Ala-D-Ala termini of bacterial cell wall precursors, which are the substrate of transglycosylase/transpeptidase. We have characterized a cluster of seven genes (vanSRJKHAX) in Streptomyces coelicolor that confers inducible, high-level vancomycin resistance. vanHAX are orthologous to genes found in vancomycin resistant enterococci that encode enzymes predicted to reprogramme peptidoglycan biosynthesis such that cell wall precursors terminate in D-Ala-D-Lac rather than D-Ala-D-Ala. vanR and vanS encode a two-component signal transduction system that mediates transcriptional induction of the seven van genes. vanJ and vanK are novel genes that have no counterpart in previously characterized vancomycin resistance clusters from pathogens. VanK is a member of the Fem family of enzymes that add the cross-bridge amino acids to the stem pentapeptide of cell wall precursors, and vanK is essential for vancomycin resistance. The van genes are organized into four transcription units, vanRS, vanJ, vanK and vanHAX, and these transcripts are induced by vancomycin in a vanR-dependent manner. To develop a sensitive bioassay for inducers of the vancomycin resistance system, the promoter of vanJ was fused to a reporter gene conferring resistance to kanamycin. All the inducers identified were glycopeptide antibiotics, but teicoplanin, a membrane anchored glycopeptide, failed to act as an inducer. Analysis of mutants defective in the vanRS and cseBC cell envelope signal transduction systems revealed significant cross-talk between the two pathways. PMID- 15130129 TI - Analysis of the 5S rRNA gene promoter from Acanthamoeba castellanii. AB - The promoter region of the Acanthamoeba 5S rRNA gene was analysed by in vitro transcription of several 5' and 3' deletion and substitution mutants, as well as a series of linker scanning mutants. The promoter consists of three sequence regions contained entirely within the gene; two of these correspond to the A and C boxes that bind TFIIIA, found in the genes from other genera. In addition, a region immediately 3' to the transcription start site has a strong effect on initiation efficiency. No strict requirement was found for specific sequences 5' to the transcription start site. Substitution of a consensus TATA box at -29 had only a modest effect on transcription, and deletion or substitution of sequences between -15 and -10 as well as -34 and -21 was only modestly more active than the wild-type template. Analysis of 3' deletions sets the 3' end-point of the promoter between +79 and +97, and demonstrates the importance of a T-rich region in transcription termination. Taken together, these results suggest that promoter elements within the Acanthamoeba 5S RNA gene are somewhat redundant, with the exception of a sequence between +50 and +60, which functions in binding TFIIIA. Remarkably, polymerase chain reaction product templates containing only non specific 5' ends between -6 and +1 relative to the transcription start site are fully functional, demonstrating that no external DNA scaffold is needed for TFIIIB and RNA polymerase III binding, and that productive initiation can be mediated solely by protein-DNA interactions within the coding region of the 5S gene. PMID- 15130130 TI - Transcriptional fidelity and proofreading in Archaea and implications for the mechanism of TFS-induced RNA cleavage. AB - We have addressed the question whether TFS, a protein that stimulates the intrinsic cleavage activity of the archaeal RNA polymerase, is able to improve the fidelity of transcription in Methanococcus. Using non-specific transcription experiments, we could demonstrate that misincorporation of non-templated nucleotides is reduced in the presence of TFS. A more detailed analysis revealed that elongation complexes containing a misincorporated nucleotide were arrested, but could be reactivated by TFS. RNase as well as exonuclease III footprinting experiments demonstrated that this arrest was not combined with extended backtracking. Analysis of paused elongation complexes demonstrated that TFS is able to induce a cleavage resynthesis cycle in such complexes, which resulted in the accumulation of dinucleotides corresponding to the last two nucleotides of the transcript. Further analysis of cleavage products revealed that, even under conditions that strongly promote misincorporation, still 50% of the released dinucleotides were correctly incorporated. Therefore, we assume that pausing of elongation complexes is an important determinant of TFS-induced RNA cleavage from the 3' end. As the incorporation rate of wrong nucleotides is about 700-fold reduced, it is possible that this delay also provides an appropriate time window for cleavage induction in order to maintain transcriptional fidelity by preventing misincorporation. PMID- 15130131 TI - Molecular analysis of the key cytokinetic components of cyanobacteria: FtsZ, ZipN and MinCDE. AB - Using a bacterial two-hybrid system and a combination of in vivo and in vitro assays that take advantage of the green fluorescent reporter protein (GFP), we have investigated the localization and the protein-protein interaction of several key components of the cytokinetic machinery of cyanobacteria (i.e. the progenitor of chloroplast). We demonstrate that (i) the ftsZ and zipN genes are essential for the viability of the model cyanobacterium Synechocystis sp. PCC 6803, whereas the minCDE cluster is dispensable for cell growth; (ii) the GTP-binding domain of FtsZ is crucial to FtsZ assembly into the septal ring at mid-cell; (iii) the Z ring of deeply constricted daughter cells is oriented perpendicularly to the mother Z-ring, showing that Synechocystis divides in alternating perpendicular planes; (iv) the MinCDE system affects the morphology of the cell, as well as the position and the shape of FtsZ structures; and (v) MinD is targeted to cell membranes in a process involving its C-terminal amphipathic helix, but not its ATP-binding region. Finally, we have also characterized a novel Z-interacting protein, ZipN, the N-terminal DnaJ domain of which is critical to the decoration of the Z-ring, and we report that this process is independent of MinCDE. PMID- 15130132 TI - An amino-terminal domain of Enterococcus faecalis aggregation substance is required for aggregation, bacterial internalization by epithelial cells and binding to lipoteichoic acid. AB - Aggregation substance (AS), a plasmid-encoded surface protein of Enterococcus faecalis, plays important roles in virulence and antibiotic resistance transfer. Previous studies have suggested that AS-mediated aggregation of enterococcal cells could involve the binding of this protein to cell wall lipoteichoic acid (LTA). Here, a method to purify an undegraded form of Asc10, the AS of the plasmid pCF10, is described. Using this purified protein, direct binding of Asc10 to purified E. faecalis LTA was demonstrated. Equivalent binding of Asc10 to LTA purified from INY3000, an E. faecalis strain that is incapable of aggregation, was also observed. Surprisingly, mutations in a previously identified aggregation domain from amino acids 473 to 683 that abolished aggregation had no effect on LTA binding. In frame deletion analysis of Asc10 was used to identify a second aggregation domain located in the N-terminus of the protein from amino acids 156 to 358. A purified Asc10 mutant protein lacking this domain showed reduced LTA binding, while a purified N-terminal fragment from amino acids 44-331 had high LTA binding. Like the previously described aggregation domain, the newly identified Asc10((156-358)) aggregation domain was also required for efficient internalization of E. faecalis into HT-29 enterocytes. Thus, Asc10 possess two distinct domains required for aggregation and eukaryotic cell internalization: an N-terminal domain that promotes binding to LTA and a second domain located near the middle of the protein. PMID- 15130133 TI - Enhancement and rescue of target capture in Tn10 transposition by site-specific modifications in target DNA. AB - The bacterial transposon Tn10 inserts preferentially into specific target sequences. This insertion specificity appears to be linked to the ability of target sites to adopt symmetrically positioned DNA bends after binding the transposition machinery. Target DNA bending is thought to permit the transposase protein to make additional contacts with the target DNA, thereby stabilizing the target complex so that the joining of transposon and target DNA sequences can occur efficiently. In the current work, we have asked whether the introduction of a discontinuity in a target DNA strand, a modification that is expected to make it easier for a DNA molecule to bend, can enhance or rescue target capture under otherwise suboptimal reaction conditions. We show that either a nick or a missing phosphate specifically at the site of reaction chemistry increases the ability of various target DNAs to form the target capture complex. The result suggests that the bends in the target DNA are highly localized and include the scissile phosphates. This raises the possibility that strand transfer is mechanistically linked to target capture. We have also identified specific residues in the target DNA and in transposase that appear to play an important role in target DNA bending. PMID- 15130134 TI - Quantitative proteomics of the human malaria parasite Plasmodium falciparum and its application to studies of development and inhibition. AB - The ability to measure accurately comparative levels of protein expression after drug challenge, metabolic stress, developmental programming or other perturbation represents one of the most important goals in post-genomics malaria research. We describe here a simple and robust quantitative methodology that is ideally suited to in vitro experiments designed to study changes in the proteome of the most important of the human parasites, the lethal species Plasmodium falciparum. The metabolic labelling technique we have developed uses parasite uptake of heavy isotope-containing isoleucine during normal growth followed by two-dimensional separation of individual proteins and mass spectrometry. The method is applicable to essentially each of the approximately 5300 proteins of P. falciparum predicted from the completed genome sequence, permitting facile identification and accurate comparative quantification of labelled peptides from any of these proteins synthesized by in vitro cultures subjected to different stimuli. We demonstrate its application to the study of cell cycle changes, where we observe divergent patterns of protein and reported transcript levels indicative of modulation at the translational level. Our data also provide evidence for significant levels of post-translational modification in the parasite, and we measure differences among variants of phosphoethanolamine N-methyltransferase and actin-I across the cell cycle. We have also monitored parasite responses to equipotent doses of the clinical antimalarial inhibitors pyrimethamine and tetracycline and observed differential effects for a number of proteins unrelated to likely targets of these drugs. PMID- 15130135 TI - Regulation of type III secretion in Bordetella. AB - The BvgAS virulence control system regulates the expression of type III secretion genes in Bordetella subspecies that infect humans and other mammals. We have identified five open reading frames, btrS, btrU, btrX, btrW and btrV, that are activated by BvgAS and encode regulatory factors that control type III secretion at the levels of transcription, protein expression and secretion. The btrS gene product bears sequence similarity to ECF (extracytoplasmic function) sigma factors and is required for transcription of the bsc locus. btrU, btrW and btrV encode proteins predicted to contain PP2C-like Ser phosphatase, HPK (His protein kinase)-like Ser kinase and STAS anti-sigma factor antagonist domains, respectively, which are characteristic of Gram-positive partner switching proteins in Bacillus subtilis. BtrU and BtrW are required for secretion of proteins that are exported by the bsc type III secretion system, whereas BtrV is specifically required for protein synthesis and/or stability. Bordetella species have thus evolved a unique cascade to differentially regulate type III secretion that combines a canonical phosphorelay system with an ECF sigma factor and a set of proteins with domain signatures that define partner switchers, which were traditionally thought to function only in Gram-positive bacteria. The presence of multiple layers and mechanisms of regulation most likely reflects the need to integrate multiple signals in controlling type III secretion. The bsc and btr loci are nearly identical between broad-host-range and human-specific Bordetella. Comparative analysis of Bordetella subspecies revealed that, whereas bsc and btr loci were transcribed in all subspecies, only broad-host-range strains expressed a functional type III secretion system in vitro. The block in type III secretion is post-transcriptional in human-adapted strains, and signal recognition appears to be a point of divergence between subspecies. PMID- 15130136 TI - Plasmid partitioning and the spreading of P1 partition protein ParB. AB - Bacterial plasmids of low copy number, P1 prophage among them, are actively partitioned to nascent daughter cells. The process is typically mediated by a pair of plasmid-encoded proteins and a cis-acting DNA site or cluster of sites, referred to as the plasmid centromere. P1 ParB protein, which binds to the P1 centromere (parS), can spread for several kilobases along flanking DNA. We argue that studies of mutant ParB that demonstrated a strong correlation between spreading capacity and the ability to engage in partitioning may be misleading, and describe here a critical test of the dependence of partitioning on the spreading of the wild-type protein. Physical constraints imposed on the spreading of P1 ParB were found to have only a minor, but reproducible, effect on partitioning. We conclude that, whereas extensive ParB spreading is not required for partitioning, spreading may have an auxiliary role in the process. PMID- 15130137 TI - Identification, molecular biotyping and ultrastructural studies of bacterial communities isolated from two damaged frescoes of St Damian's Monastery in Assisi. AB - AIM: To investigate the composition of the microbial community in biodeterioration of two frescoes in St Damian's Monastery in Assisi. METHODS AND RESULTS: A total of 1292 colonies were isolated from the most deteriorated parts, analysed by microbiological, biomolecular and ultrastructural techniques, and taxonomically classified. Molecular biotyping of Staphylococcus cohnii colonies, one of the most prevalent bacterial species, showed a very restricted genome diversity while Bacillus licheniformis were very homogeneous by RFLP, tDNA-PCR and random-amplified polymorphic DNA. Electron microscopy confirmed heterogeneity of the bacterial population in the different sampling areas. CONCLUSIONS: Several of the identified species are widespread in the soil or saprophytes of human skin. Although unable to demonstrate that they are involved in biodeterioration, they may represent trophic elements contributing to fungi-related chromatic alterations when adequate environmental conditions occur. Deterioration may in part be prevented or controlled by adequate air filtering or conditioning of the room. PMID- 15130138 TI - Surface alteration of Saccharomyces cerevisiae induced by thymol and eugenol. AB - AIMS: This study aims to bring some information about the mechanism of the fungicidal action of thymol and eugenol; phenolic major components of thyme and clove essential oils respectively. Saccharomyces cerevisiae was used as yeast model. METHODS AND RESULTS: Treatment of yeast cells with these components led to their lysis as shown by the release of substances absorbing at 260 nm. In addition, scanning electron microscope observations revealed that the surface of the treated cells was significantly damaged. CONCLUSIONS: Antifungal activity of thymol and eugenol involve alteration of both membrane and cell wall of the yeast. SIGNIFICANCE AND IMPACT OF THE STUDY: This work is a preliminary contribution aiming to develop a new generation of efficient and natural antifungal agents. PMID- 15130139 TI - Characterisation of autolytic enzymes in Lactobacillus pentosus. AB - AIMS: To characterize autolysis and autolytic system of the lactic acid bacterium Lactobacillus pentosus. METHODS AND RESULTS: Autolysis of nine Lact. pentosus strains was evaluated in buffer solution. Their peptidoglycan hydrolase profiles were examined by renaturing SDS-PAGE and revealed two major activity bands at 58 and 112 kDa. Specificity analysis indicated the presence of at least two different types of peptidoglycan hydrolase activities in Lact. pentosus 1091. CONCLUSIONS: Autolysis of Lact. pentosus was shown to be strain dependent and involvement of at least two different autolysins was evidenced. SIGNIFICANCE AND IMPACT OF THE STUDY: The autolytic system of Lact. pentosus was characterized for the first time and the data obtained could be used in the selection of strains of technological interest. PMID- 15130140 TI - Detection and quantification of Aspergillus ochraceus in green coffee by PCR. AB - AIMS: The aim of this study was to detect and quantify DNA of the ochratoxinogenic fungus Aspergillus ochraceus in green coffee and to compare the results with the ochratoxin A content of naturally contaminated samples. METHODS AND RESULTS: A DNA extraction protocol based on a combination of ultrasonification and a commercial kit was tested for the recovery of fungal DNA. PCR and real-time PCR protocols were established for the detection of A. ochraceus. Sensitivity of the PCR was checked by the addition of inoculated green coffee and pure fungal DNA to uncontaminated green coffee samples. The A. ochraceus DNA content of 30 naturally contaminated green coffee samples was determined and compared with the ochratoxin A concentrations. CONCLUSIONS: Aspergillus ochraceus can be rapidly and specifically detected in green coffee by PCR. A positive correlation between the ochratoxin A content and the DNA quantity was established. SIGNIFICANCE AND IMPACT OF THE STUDY: This work offers a quick alternative to the conventional mycological detection and quantification of A. ochraceus in green coffee. PMID- 15130141 TI - The effect of lipids on the adherence of axillary aerobic coryneform bacteria. AB - AIMS: A wide range of lipids are present on the skin surface of human beings and bacterial lipases are known to modify them. The microflora of the underarm (axilla) is often dominated by aerobic coryneforms and whilst many require lipids for growth, they appear not to be utilized as carbon sources. The aim of this study was to investigate the adherence aerobic coryneforms to lipids present on the skin surface of the human axilla to determine whether they contribute to colonization of the skin. METHODS: Aerobic coryneforms were grown in a defined synthetic medium in the presence of (14)C-glucose to produce radio-labelled cells. Adherence to lipids was tested using a thin layer chromatography plate based assay. SIGNIFICANCE AND IMPACT OF THE STUDY: The mechanism of bacterial adhesion to skin lipids is unknown. The results of this study show that a significant proportion of cutaneous aerobic coryneform isolates from the axilla interact with skin lipids resulting in increased adherence, which may contribute to skin colonization. PMID- 15130142 TI - Molecular confirmation of Enterococcus faecalis and E. faecium from clinical, faecal and environmental sources. AB - AIMS: The genus Enterococcus includes opportunistic pathogens such as E. faecalis and E. faecium, and is also used to assess water quality. Speciation of enterococci in environmental studies can be particularly problematic, therefore protocols for unambiguous, DNA-based analysis could receive wide use in applications ranging from water quality monitoring to microbial source tracking. The goal of this work was to investigate the usefulness of PCR for speciation of putative, biochemically identified E. faecalis and E. faecium isolated from water, faeces and sewage. METHODS AND RESULTS: Putative enterococci (n = 139) were isolated on mEI agar from dog, human, gull and cow faeces, and from sewage, freshwaters and marine waters. A total of 128 isolates passed standard physiological tests for the genus, and were speciated by the API 20 Strep (APIStrep) biochemical test system. 42.2% were identified as E. faecalis, and all were confirmed by PCR. 19.5% were biochemically identified as E. faecium, but only seven were PCR-positive. CONCLUSIONS: The 16S rDNA of PCR-positive and PCR negative E. faecium, including isolates that were inconclusively identified by APIStrep, was sequenced. All formed a monophyletic clade with E. faecium sequences in Genbank. SIGNIFICANCE AND IMPACT OF THE STUDY: Biochemical identification of E. faecalis agreed 100% with PCR assays, therefore a simple protocol of isolation on mEI followed by PCR should be useful for environmental studies. Discrepancies among biochemical identification, PCR confirmation and DNA sequencing were noted for E. faecium, indicating that routine isolation/identification of E. faecium from environmental samples is a much more difficult task. PMID- 15130143 TI - Simple staining detects ultrastructural and biochemical differentiation of vegetative hyphae and fruit body initials in colonies of Pleurotus pulmonarius. AB - AIMS: To know the ultrastructural and biochemical differences of vegetative hyphae and fruit body initials in colonies of Pleurotus pulmonarius. METHODS AND RESULTS: Feulgen reagent was used to detects differentiation of hyphae. The intracellular laccases, proteases and beta-1,3-glucanases activity, content of cytoplasmic protein, glycogen and glucans in the cell wall were evaluated in hyphae of fruit body initials and in vegetative hyphae. The thickness of hyphal walls of the vegetative hyphae was also evaluated. Substantial biochemical changes were observed in hyphae of different zones of the fruiting colony. Hyphae at the periphery had thinner walls than in the centre of the colony. CONCLUSION, SIGNIFICANCE AND IMPACT OF THE STUDY: Staining correlated with the enzymatic activity, protein, glycogen and glucans, in mycelium and in fruit body initials. The implications are that hyphal maturity in P. pulmonarius involves storage of glucans, in part at least, in the form of a thickened hyphal wall. PMID- 15130144 TI - Production of slime polysaccharide by EHEC and STEC as well as the influence of culture conditions on slime production in Escherichia coli O157:H7. AB - AIMS: To quantify the slime polysaccharide, composed of colanic acid (CA), produced by enterohaemorrhagic and Shiga toxin-producing Escherichia coli (EHEC and STEC) and to determine the influence of culture conditions on CA production in E. coli O157:H7. METHODS AND RESULTS: The study examined the amounts of CA produced by EHEC and STEC, and evaluated the production of CA in E. coli O157:H7 as influenced by medium pH and incubation temperatures. The results indicated that the amounts of CA produced by EHEC and STEC vary to a great extent and CA production in E. coli O157:H7 is influenced by the tested culture conditions. CONCLUSIONS: The abilities of EHEC and STEC to produce CA differ. Medium pH and incubation temperature are among the important factors affecting CA production in E. coli O157:H7. SIGNIFICANCE AND IMPACT OF THE STUDY: Slime polysaccharide can affect the abilities of E. coli O157:H7 cells to combat environmental stress. This study contributes to a better understanding of the physiological factors influencing slime polysaccharide production in EHEC and STEC. PMID- 15130145 TI - Occurrence and clonal relatedness of sec/tst-gene positive Staphylococcus aureus isolates of quartermilk samples of cows suffering from mastitis. AB - AIMS: To investigate the prevalence of sec/tst-gene positive Staphylococcus aureus in bovine mastitis and to get information about the clonal relatedness of these clinical isolates. METHODS AND RESULTS: A total of 533 Staph. aureus strains isolated from bovine mastitic quartermilk samples at 493 randomized dairy farms in Hessia, Germany, from January 1997 until June 1998 were examined for enterotoxin C (sec) gene and toxic shock syndrome toxin (tst) gene by multiplex polymerase chain reaction. Fifty-three (9.3%) of the strains were sec/tst-gene positive. Phenotypic TSST-1 production was found in all positive strains by reversed passive latex agglutination test. With DNA macrorestriction analysis, sec/tst-gene positive strains were divided into five different macrorestriction types. Type I (10 isolates) and III (40 isolates) were found to be the predominant types in terms of frequency of isolation in the investigated area. These DNA macrorestriction types differed in only two bands in the 500 and 270 bp region. CONCLUSIONS: Closely related Staph. aureus strains seem to be responsible for an unusual large proportion of bovine mastitis cases in geographically widely distinct locations. SIGNIFICANCE AND IMPACT OF THE STUDY: This is one of the first reports about the relatedness of sec/tst-gene positive Staph. aureus clinical isolates from bovine mastitis. PMID- 15130146 TI - Thermal and chemical resistance of Lactobacillus casei and Lactobacillus paracasei bacteriophages. AB - AIMS: The survival of two collection Lactobacillus casei and L. paracasei bacteriophages when subjected to thermal and chemical treatments was investigated. METHODS AND RESULTS: Thermal resistance was evaluated by heating phage suspensions at 63, 72 and 90 degrees C in three different media [Tris magnesium gelatin (TMG) buffer: 10 mmol l(-1) Tris-Cl, 10 mmol l(-1) MgSO(4) and 0.1% w/v gelatin; Man Rogosa Sharpe (MRS) broth and reconstituted nonfat dry skim milk (RSM)]. A marked heat sensitivity was evident in both phages, as 15 min at 72 degrees C was enough to completely inactivate (6 log(10) reduction) them. No clear influence was demonstrated by the suspension media. The phages also showed similar resistance to biocides. Peracetic acid and sodium hypochlorite (800 ppm) were the most effective ones, destroying the phages within 5 min. Concentrations of 75 and 100% ethanol were not suitable to inactivate phage particles even after 45 min. Isopropanol did not show an effect on phage viability. CONCLUSIONS: The data obtained in this work are important to design more effective control procedures in order to inactivate phages in dairy plants and laboratories. SIGNIFICANCE AND IMPACT OF THE STUDY: This work will contribute to enhance the background knowledge about phages of probiotic bacteria. PMID- 15130147 TI - Inactivation of Campylobacter jejuni by high hydrostatic pressure. AB - AIMS: To investigate the response of Campylobacter jejuni ATCC 35919 and 35921 to high pressure processing (HPP) while suspended in microbiological media and various food systems. METHODS AND RESULTS: Campylobacter jejuni 35919 and 35921 were subjected to 10-min pressure treatments between 100 and 400 MPa at 25 degrees C suspended in Bolton broth, phosphate buffer (0.2 m, pH 7.3), ultra-high temperature (UHT) whole milk, UHT skim milk, soya milk and chicken puree. The survivability of C. jejuni was further investigated by inoculated pack studies. HPP at 300-325 MPa for 10 min at 25 degrees C was sufficient to reduce viable numbers of both strains to below detectable levels when cells were pressurized in Bolton broth or phosphate buffer. All food products examined offered a protective effect in that an additional 50-75 MPa was required to achieve similar levels of inactivation when compared with broth and buffer. Inoculated pack studies showed that the survivability of C. jejuni following pressurization improved with decreasing post-treatment storage temperature. SIGNIFICANCE AND IMPACT OF THE STUDY: These data demonstrated that HPP at levels of or = 13). There was no significant association between length of stay (1 2 days vs. > or = 5 days, 3-4 days vs. > or = 5 days) and primary outcome measures in univariable analyses of the 1989 Survey, or multivariable analyses of the 1994 and 2000 Surveys adjusting for relevant social and obstetric factors. For stays of 3-4 days, the adjusted odds ratio for formula feeding at 6 weeks was 1.35 [95% CI 0.9, 1.9] in 1994 and 1.22 [95% CI 0.9, 1.7] in 2000. The confidence intervals are compatible with a very small reduction or a large increase in formula feeding, neither reaching statistical significance. For depressive symptoms at 5-7 months postpartum (EPDS score > or = 13), the adjusted odds ratio for women staying 3-4 days was 0.96 [95% CI 0.7, 1.4] in 1994 and 0.90 [95% CI 0.6, 1.3] in 2000. These confidence intervals are compatible with a 30-40% reduction or a 30-40% increase in odds of depressive symptoms. Based on these findings shorter length of stay does not appear to have an adverse impact on breast feeding or women's emotional well-being. Testing early discharge policies in well-designed randomised trials remains a priority for developing stronger evidence to inform practice. PMID- 15130161 TI - Cerebral palsy of post-neonatal origin: characteristics and risk factors. AB - We aimed to study the rates and trends over time of children with cerebral palsy (CP) of post-neonatal origin (arising more than 28 days after birth, and before the age of 25 months), to examine their aetiology and associated significant risk factors, and to compare them with other CP cases. Children with post-neonatal CP born 1976-90 were identified from a European database and seven registers were included (Surveillance of Cerebral Palsy in Europe collaboration). Using a previously published classification it was possible to allocate an aetiology to 99% of cases. The prevalence rate of post-neonatal CP was 1.26 per 10 000 live births and a significant decrease was observed over the period 1976-90 (P = 0.011). Infection accounted for 50%, vascular episodes for 20% and head injury for 18% of the cases. Although there has been little change in the profile of underlying causes in this period, within the infection group, a significant downward trend was observed for Reye's syndrome (P < 0.001) and non-central nervous system (non-CNS) infection (P = 0.004), but not for meningitis/encephalitis. There was evidence of some increased risk of post neonatal CP among children with low birthweight (<2500 g) (P < 0.001). Overall children with CP of post-neonatal origin showed a more severe functional pattern than non-post-neonatal CP children. In order to ascertain the impact of public health and other preventive measures aimed at reducing the frequency of brain injury in the first 2 years after birth, it is necessary to continue to monitor the frequency and characteristics of children with post-neonatal CP into the 1990s. PMID- 15130162 TI - The Aberdeen Children of the 1950s cohort study: background, methods and follow up information on a new resource for the study of life course and intergenerational influences on health. AB - In this paper we introduce and describe in detail an addition to the UK's population-based resources for the investigation of biological and social influences on health across the life course and between generations: the Aberdeen Children of the 1950s study. We also provide an account of postwar Aberdeen when study members were growing up, report on findings of analyses of data from the original survey on which this study is based and its follow-up, assess the strengths and limitations of the study, and outline current and future research directions. This cohort comprises individuals born in Aberdeen, Scotland (UK) between 1950 and 1956, and is derived from 15 thousand subjects who took part in the Aberdeen Child Development Survey, a cross-sectional study of 'mental subnormality' (learning disability) in a population of all children who were attending Aberdeen primary schools in December 1962. Data collection included information on birthweight, gestational age, childhood height and weight, tests of cognition and behavioural disorder, and a range of multilevel socio-economic indicators. In 1998 we began the process of revitalising this cohort (now termed the Aberdeen Children of the 1950s study). We have been successful in ascertaining the current vital status and whereabouts of 98.5% of a target population of 12 150 subjects (6276 males, 5874 females) with full baseline data. The large majority (81%) of study participants still reside in Scotland and many (73%) have remained in the Grampian region which incorporates Aberdeen. At the present time, a total of almost 500 subjects are known to have died. Linkages to hospital admissions and other health endpoints captured through the Scottish Morbidity Records system have been completed. This includes an intergenerational linkage to approximately eight thousand deliveries in Scotland occurring to female members of the study population. A postal questionnaire to all traced surviving cohort members has also been distributed. PMID- 15130163 TI - Training the next generation of reproductive, perinatal and paediatric epidemiologists. PMID- 15130164 TI - The use of volume expansion in preterm infants. PMID- 15130165 TI - Correctable visual impairment in older people: a major unmet need. AB - This review seeks to determine the prevalence of correctable visual impairment (VI) in older people in the UK, to discover what proportion of these cases are undetected, to suggest reasons for the poor detection and to make recommendations for improving the detection. To establish the context of these issues, the review will also touch on the general prevalence and causes of VI in older people in developed countries and on the impact of VI in older people. Typically, studies suggest that VI affects about 10% of people aged 65-75, and 20% of those aged 75 or older. There is a strong relationship between impaired vision in older people and both reduced quality of life and increased risk of accidents, particularly falls. The literature suggests that those with low vision are about two times more likely to have falls than fully sighted people, and the annual UK cost of treating falls directly attributable to VI is pound 128 million. The literature on the prevalence of undetected reduced vision in older people reveals that between 20 and 50% of older people have undetected reduced vision. The majority of these people have correctable visual problems (refractive errors or cataract). It is particularly startling that, in 'developed countries', between 7 and 34% of older people have VI that could simply be cured by appropriate spectacles. The reasons why so many cases of treatable VI remain untreated are discussed, and suggestions are made for improving the detection of these cases. We conclude that there should be better publicity encouraging older people to attend for regular optometric eye examinations. A complementary approach is annual visual screening of the elderly, possibly as part of GPs annual health check on people aged 75 years and older. Recommendations are made for evaluating new approaches to screening and for improving the management of cases detected by screening. PMID- 15130166 TI - A novel visual analogue scale (VAS) device: an instrument based on the VAS designed to quantify the subjective visual experience. AB - PURPOSE: To evaluate a novel device intended to facilitate the direct quantification of the subjective visual experience. The design principle is based on that of the visual analogue scale (VAS) technique but obviates the need for subsequent measurement unlike the administration of the conventional paper VAS. METHODS: The visual experience of 134 normally sighted 17-40-year-olds was quantified using (in randomised sequence) a paper VAS and the novel VAS device. A notional 100-point scale was utilised in either case, and the extreme descriptive anchors for both instruments were 'dreadful' and 'perfect'. RESULTS: The degree of clinical agreement between the two alternative VAS techniques was very high. A small bias (mean = +0.7 VAS units: 95% confidence interval 0.3-1.2 units) towards the paper VAS was evident in these data, an outcome of no clinical significance or impediment as regards the substitution of the novel device for the conventional paper-based approach. CONCLUSIONS: The moving of a bead along a wire is shown to be a clinically and statistically reliable alternative procedure to the pen-marking of a conventional paper VAS for the evaluation of the recent subjective visual experience, provided that the same minimum/maximum descriptive anchors are used for either instrument. The instant quantification of the VAS score afforded by the novel technique introduced here broadens the potential clinical application of this technique. PMID- 15130167 TI - UTP and diadenosine tetraphosphate accelerate wound healing in the rabbit cornea. AB - Nucleotides are naturally occurring substances present in tear film that can stimulate tear secretion in animals and humans. We investigated whether certain nucleotides can affect the rate of wound healing in the cornea of white rabbits. In the absence of any added compound, the rate of healing was 72.4 +/- 2.2 microm h(-1). Of all the tested nucleotides, UTP and Ap(4)A were the most active ones, maximally increasing the rate of healing to 121.6 +/- 3.7 and 93.7 +/- 3.2 microm h(-1), respectively. Responses to UTP were dose dependent. UTP had a pD(2) value of 8.9 +/- 0.1 (EC(50): 1.25 nM). P2 purinoceptor antagonists such as suramin and reactive blue-2, inhibited the effect of UTP indicating the involvement of P2Y receptors. Mitogen-activated protein kinase (MAPK) cascade inhibitors also abolished the effects of UTP, suggesting that P2Y receptors are coupled to the MAPK cascade, and that this is involved in controlling the rate of epithelial cell migration. PMID- 15130168 TI - Evidence for transient forces/strains at the optic nerve head in myopia: repeated measurements of the Stiles-Crawford effect of the first kind (SCE-I) over time. AB - Probable transient changes in photoreceptor alignments, inferred from the measurements of the Stiles-Crawford effect of the first kind (SCE - I), were demonstrated in myopic eyes with elongated axial length (regardless of the magnitude of refractive error) at three retinal locations; the fovea (point of fixation), and 22 degrees and 27 degrees in the nasal retina. The changes were much bigger at 22 degrees and 27 degrees in the nasal retina (which are located beyond the optic nerve head) than at the fovea. These transient effects were revealed by repeatedly testing the same retinal locations over a period of time. Time intervals between the subsequent/repeated measurements ranged from less than an hour to several months. In some locations, the changes were recorded in less than an hour. Collectively, these were very meaningful changes. Generally bigger effects were recorded in the horizontal SCE - I than in the vertical SCE - I. PMID- 15130169 TI - Stimulating human accommodation without changes in focus. AB - PURPOSE: Inspired by the finding in chickens that preferential stimulation of the ON retinal system suppresses myopia induced by negative spectacle lens wear and that stimulation of the OFF system suppresses the hyperopia induced by positive lens wear, we sought to determine whether stimulation of the ON-OFF retinal systems could drive directional accommodation responses in humans. If emmetropisation and accommodation use similar image processing algorithms, more accommodation would be expected with OFF stimulation. METHODS: Accommodation responses were measured while viewing a computer-generated pattern designed to stimulate the ON-OFF systems. The stimulus comprised a rectangular field (12 x 9.5 cm) on a black background filled with 196 discs (diameters: 0.4-1.0 cm). These were presented on an LCD monitor in a dark room at a viewing distance of 55 cm (1.8 D). Thirteen subjects aged 21-37 years took part. The individual discs had saw-tooth shaped temporal luminance profiles with the same time period but with random phases with respect to each other, so that the mean brightness of the stimulus was constant. To eliminate accommodation responses based on other cues (i.e. proximity) a 0.5 mm artificial pupil was used to open the accommodation loop. Refraction in the vertical pupil meridian was continuously recorded with an infrared photorefractor (the PowerRefractor). To verify that computer-based stimuli presented within our experimental design were effective in driving accommodation, previously studied stimuli were also tested: changes in size (looming) and incremental low pass filtering. RESULTS: Preferential stimulation of the ON or OFF subsystems produced a convincing depth illusion in all subjects (which was psychophysically confirmed in four subjects). Although the stimulus appeared to move in depth it did not produce accommodation responses that were consistent with that, i.e. the accommodation system did not appear to fluctuate in rhythm with the temporal oscillations of the stimulus. As the target appeared to loom it induced a greater accommodation response then when it appeared to recede. The looming target produced changes in the accommodation response in nine of 13 subjects that were consistent with its perceived change in proximity (although the target did not actually move in depth). Incremental low pass filtering produced non-directional drifts of accommodation in all subjects. Combinations of the stimuli (i.e. looming and low pass filtering, ON/OFF and looming) were not more effective stimuli to accommodation. After removal of the artificial pupil (closed loop conditions), accommodation was no longer induced with any of these stimuli. CONCLUSIONS: Although the preferential ON or OFF stimulation produced a pronounced illusion of motion in depth despite constant average brightness, proximal accommodation was induced in only one subject. Therefore, the ON/OFF stimulation appeared to have only minor input into proximal accommodation. Potential inputs into reflex accommodation need to be defined in further studies. PMID- 15130170 TI - Effect of storage temperatures and time on the efficacy of multipurpose solutions for contact lenses. AB - PURPOSE: To determine the effect of storage time and temperature on the efficacy of four multipurpose solutions for soft contact lenses. METHOD: Aliquots of multipurpose solutions (OPTI-FREE Express, ReNu MultiPlus, COMPLETE and SOLO care) stored at different temperatures over a 3-month period, were challenged with contact lens-related ocular pathogens, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans. RESULTS: The results showed that OPTI-FREE Express had the best activity against Ps. aeruginosa at all temperatures; ReNu MultiPlus performed well at 25 degrees C; COMPLETE barely achieved activity requirements at all temperatures, and lost efficacy after 2 months. SOLO-care maintained its activity best against Ps. aeruginosa at 30 degrees C. Storage at fridge temperature reduced activity of all solutions. Regardless of storage temperature, activities of all solutions against S. aureus markedly decreased by 2 months. Only OPTI-FREE Express met FDA requirements against C. albicans. CONCLUSION: Performance of multipurpose solutions is affected by time and temperature of storage. Contact lens users should be aware that the efficacy of opened solutions may not be sustained for as long as 3 months. Manufacturers should reconsider their recommendations to further safeguard the ocular health of contact lens wearers. PMID- 15130171 TI - Chromatic and luminance losses with multiple sclerosis and optic neuritis measured using dynamic random luminance contrast noise. AB - We measured thresholds for detecting changes in colour and in luminance contrast in observers with multiple sclerosis (MS) and/or optic neuritis (ON) to determine whether reduced sensitivity occurs principally in red-green or blue-yellow second stage chromatic channels or in an achromatic channel. Colour thresholds for the observers with MS/ON were higher in the red-green direction than in the blue yellow direction, indicating greater levels of red-green loss than blue-yellow loss. Achromatic thresholds were raised less than either red-green or blue-yellow thresholds, showing less luminance-contrast loss than chromatic loss. With the MS/ON observers, blue-yellow and red-green thresholds were positively correlated but increasing impairment was associated with more rapid changes in red-green thresholds than blue-yellow thresholds. These findings indicate that demyelinating disease selectively reduces sensitivity to colour vision over luminance vision and red-green colours over blue-yellow colours. PMID- 15130172 TI - Self-optimised vision correction with adaptive spectacle lenses in developing countries. AB - It is estimated by the World Health Organization that about 1 billion people in the developing world would benefit immediately from distance vision and near vision correction if it were available to them. Here we address this problem and provide a solution by correcting vision in the field with adaptive liquid-filled variable focus lenses. We describe the details of the lens and present the results of a series of experiments performed in Ghana, Nepal, Malawi and South Africa that demonstrate that it is possible by simple means to self-determine and obtain correct refraction using such lenses. PMID- 15130173 TI - A quantitative description of lens eye morphology and its implications. AB - If it is assumed that the differences between eyes constitute adaptations to different visual environments, then it should be possible to identify the visual environments for which the eye is optimised from these differences. Two different classes of simple eye shapes can be distinguished from the ratio of maximum pupil size to the axial length of an eye. The quantitative description of the two different eye shapes can be used to identify the relative importance of image illumination and resolution to an animal from a cross-section of its eye. PMID- 15130174 TI - The amplitude of accommodation in 6-10-year-old children - not as good as expected! AB - The aim of this study was to measure the amplitude of accommodation for junior level school children and to compare it with age-expected values. A junior level school in Goteborg, Sweden, was randomly chosen and the amplitude of accommodation among 76 children aged 6-10 years was examined using Donders' push up method. The results showed lower amplitude than expected in a large group of children. Results also showed lower amplitude than previously reported for this age group, especially under monocular conditions, which revealed an average dioptric difference from the expected value of -3.60 dioptres (D) right eye (mean 12.40 D, median 12.00 D, S.D. 3.7 D) and -3.50 D left eye (mean 12.50 D, median 12.70 D, S.D. 3.8 D) (p < 0.001 for both eyes). Consequently, we conclude that it cannot be assumed that the amplitude of accommodation is in the expected amplitude range for all children of these ages. PMID- 15130179 TI - Human pregnancy: the role of chemokine networks at the fetal-maternal interface. AB - Chemokines are multifunctional molecules initially described as having a role in leukocyte trafficking and later found to participate in developmental processes such as differentiation and directed migration. Similar events occur in pregnancy during development of the fetal-maternal interface, where there is extensive leukocyte trafficking and tissue morphogenesis, and this is accompanied by abundant chemokine expression. The relationship between chemokines, leukocytes and placental development is beginning to be delineated. During pregnancy a specialised population of maternal leukocytes infiltrates the implantation site. These leukocytes are thought to sustain the delicate balance between protecting the developing embryo/fetus and tolerating its hemiallogeneic tissues. A network of chemokine expression by both fetal and maternal components in the pregnant uterus functions in establishing this leukocyte population. Intriguingly, experiments investigating immune cell recruitment revealed the additional possibility that chemokines influence aspects of placental development. Specifically, cytotrophoblasts, the effector cells of the placenta, express chemokine receptors that can bind ligands found at key locations, implicating chemokines as regulators of cytotrophoblast differentiation and migration. Thus, as in other systems, at the fetal-maternal interface chemokines might regulate multiple functions. PMID- 15130180 TI - Methodological issues in the measurement of birth preparedness in support of safe motherhood. AB - Behavior change interventions focusing on birth preparedness for pregnant women, their husbands, and adults in the community are common components of community oriented Safe Motherhood programs in developing countries. Few studies have examined the effectiveness of these interventions, and existing studies are flawed due to study and sample design. This article highlights methodological issues that are often overlooked when measuring indicators of birth preparedness among multiple audiences for program evaluation purposes in household-based surveys. Solutions are proposed to address each of these problems in an effort to improve future research. PMID- 15130181 TI - Using civil representation to reduce delinquency among troubled youth. AB - Team Child is designed to provide civil legal representation for very troubled delinquent youth to improve their access to needed education programs, mental health services, and family services. Better provision of these services is intended to reduce delinquency and potentially avoid the long-term incarceration of these youth. This article uses an incidental truncation model to evaluate two Team Child programs in Florida. The authors find the programs not to be effective in reducing recidivism among the youth but to be effective in reducing the arrest rate of the recidivating juveniles. The results indicate a reduction of 11% to 23% of arrests after treatment for the Team Child group. PMID- 15130182 TI - The outlook is G.R.E.A.T.: What educators say about school-based prevention and the Gang Resistance Education and Training (G.R.E.A.T.) program. AB - This article reports on a survey of administrators, counselors, and teachers from middle schools involved in the National Evaluation of the Gang Resistance Education and Training (G.R.E.A.T.) program. This survey was part of a multisite evaluation that also elicited the responses of students, parents, and officers teaching the G.R.E.A.T. program. School personnel provide their views about important current issues, including their perceptions of school safety, the role of law enforcement officers in schools, and the role and effectiveness of school based prevention programs in general and the G.R.E.A.T. program in particular. Results from this survey have important implications for the plethora of prevention programs currently located in American schools. PMID- 15130183 TI - How does active parental consent influence the findings of drug-use surveys in schools? AB - This study examines the impact of passive and active parental consent procedures on the type of adolescents participating in a school-based survey examining substance use. Schools recruited from a random sample of metropolitan schools were assigned to passive or active parental consent condition. Results showed that participation rates in active consent schools were lower than in passive consent schools for junior students (60% vs. 80%) but not senior students. Although consent condition had limited impact on prevalence estimates among older students, among younger students estimates of cannabis use and ecstasy use were higher in the passive consent condition than the active consent condition. Active consent procedures introduce some degree of selection bias into studies of adolescents' substance use and may compromise the external validity of prevalence estimates produced, especially among younger students. PMID- 15130184 TI - Early father's and mother's involvement and child's later educational outcomes. AB - BACKGROUND: Few studies have investigated the individual long-term contributions that mothers and fathers make to their children's schooling. AIMS: (1) To explore the role of early father involvement in children's later educational attainment independently of the role of early mother involvement and other confounds, (2) to investigate whether gender and family structure moderate the relationship between father's and mother's involvement and child's educational attainment, and (3) to explore whether the impact of father's involvement depends on the level of mother's involvement. SAMPLE: The study used longitudinal data from the National Child Development Study. The initial sample were those 7,259 cohort members with valid data on mother involvement at age 7, father involvement at age 7, and school-leaving qualification by age 20. Of those, 3,303 were included in the final analysis. METHOD: The measures were control variables, structural factors (family structure, sibship size and residential mobility), child factors (emotional/behavioural problems, cognitive ability and academic motivation), and father's and mother's involvement. RESULTS: Father involvement and mother involvement at age 7 independently predicted educational attainment by age 20. The association between parents' involvement and educational attainment was not stronger for sons than for daughters. Father involvement was not more important for educational attainment when mother involvement was low rather than high. Not growing up in intact two-parent family did not weaken the association between father's or mother's involvement and educational outcomes. CONCLUSION: Early father involvement can be another protective factor in counteracting risk conditions that might lead to later low attainment levels. PMID- 15130185 TI - Predicting curriculum and test performance at age 7 years from pupil background, baseline skills and phonological awareness at age 5. AB - BACKGROUND: Phonological awareness tests are known to be amongst the best predictors of literacy; however their predictive validity alongside current school screening practice (baseline assessment, pupil background data) and to National Curricular outcome measures is unknown. AIM: We explored the validity of phonological awareness and orthographic measures, the Infant Index baseline assessment (Desforges & Lindsay, 1995), gender, free school dinners, and SEN status taken at age 5 in the prediction of English, Maths, and Science performance in Key Stage 1 of the National Curriculum in England. SAMPLE: 435 children from nine schools in one Local Educational Authority (LEA) were screened at entry aged 4 or 5, and at 5:8, and age 7. METHOD: We trained schools Learning Support Assistants (LSAs) to administer orthographic and phonological awareness measures. Teachers collected data on all baseline and Key Stage 1 performance measures. RESULTS: A series of 13 logistic regression analyses revealed that phonological awareness and orthographic knowledge best predicted 12 of 13 measures of performance at Key Stage 1. Additional variance was nearly always explained by SEN status. In 11 analyses the Infant Index baseline score predicted performance at age 7 and was the best predictor of maths test performance. Gender was an occasional additional predictor. CONCLUSION: School LSAs can be readily trained to administer phonological awareness measures to cohorts of 5-year-old children. Such measures used in conjunction with baseline measures significantly enhance prediction of Key Stage 1 performance. Deploying LSAs in this fashion significantly enhances a school's capacity to identify young children for whom additional support may be necessary. PMID- 15130186 TI - Educational consequences of developmental speech disorder: Key Stage 1 National Curriculum assessment results in English and mathematics. AB - BACKGROUND: Children with speech difficulties may have associated educational problems. This paper reports a study examining the educational attainment of children at Key Stage 1 of the National Curriculum who had previously been identified with a speech difficulty. AIMS: (1) To examine the educational attainment at Key Stage 1 of children diagnosed with speech difficulties two/three years prior to the present study. (2) To compare the Key Stage 1 assessment results of children whose speech problems had resolved at the time of assessment with those whose problems persisted. SAMPLE(S): Data were available from 39 children who had an earlier diagnosis of speech difficulties at age 4/5 (from an original cohort of 47) at the age of 7. A control group of 35 children identified and matched at preschool on age, nonverbal ability and gender provided comparative data. METHODS: Results of Statutory Assessment Tests (SATs) in reading, reading comprehension, spelling, writing and maths, administered to children at the end of Year 2 of school were analysed. Performance across the two groups was compared. Performance was also compared to published statistics on national levels of attainment. RESULTS: Children with a history of speech difficulties performed less well than controls on reading, spelling and maths. However, children whose speech problems had resolved by the time of assessment performed no differently to controls. Children with persisting speech problems performed less well than controls on tests of literacy and maths. Spelling performance was a particular area of difficulty for children with persisting speech problems. CONCLUSIONS: Children with speech difficulties are likely to perform less well than expected on literacy and maths SAT's at age 7. Performance is related to whether the speech problem resolves early on and whether associated language problems exist. Whilst it is unclear whether poorer performance on maths is because of the language components of this task, the results indicate that speech problems, especially persisting ones, can affect the ability to access the National Curriculum to expected levels. PMID- 15130187 TI - Can reading accuracy and comprehension be separated in the Neale Analysis of Reading Ability? AB - BACKGROUND: The Neale Analysis of Reading Ability (NARA) (Neale, 1997) is widely used in education and research. It provides measures of reading accuracy (decoding) and comprehension, which are frequently interpreted separately. AIMS: Three studies were conducted to investigate the degree to which the NARA measures could be separated. SAMPLES: British 7- and 8-year-olds participated in Study 1 (N=114) and Study 2 (N=212). In Study 3, 16 skilled and less-skilled comprehenders were identified from the Study 2 sample. METHODS: Study 1: By investigating their contribution to silent reading comprehension, the independence of NARA decoding and comprehension scores was determined. Study 2: Decoding groups matched for listening comprehension were compared on the NARA comprehension measure, and population performance was compared across listening comprehension and NARA reading comprehension. Study 3: Comprehension groups were compared on ability to answer open-ended and forced-choice questions. RESULTS: Firstly, NARA comprehension performance depended on decoding, to the extent that children with high listening comprehension ability but low decoding ability attained low NARA comprehension scores. Secondly, 32% of children who attained low NARA comprehension scores exhibited high listening comprehension. Thirdly, comprehension groups differed when assessed with open-ended questions but not when assessed with forced-choice questions. CONCLUSIONS: The NARA can underestimate the comprehension ability of children with weak decoding skills and children who have some difficulty with open-ended questions. The decoding and comprehension measures of the NARA cannot be separated. These findings have important implications for the interpretation of the measures provided by the NARA, in education and research. PMID- 15130188 TI - Teachers' task demands, students' test expectations, and actual test content. AB - BACKGROUND: Previous studies on instructional importance show that individual students and their teachers differ in the topics that they consider important in the context of an upcoming teacher-made test. AIMS: This study aimed to examine whether such differences between students' test expectations and teachers' intended task demands can be explained by the actual test content. SAMPLES: Participants were history teachers (N=19) and their 11th-grade students (N=388). METHOD: Teachers and students rated the importance of text sections that would be tested in the near future. By means of multilevel analysis, ratings were compared with the occurrence of sections in the tests. RESULTS: Although teachers considered a majority of sections as important and tested only a minority of the sections, their tests still included sections rated as unimportant. The number of such discrepancies, however, was relatively small. Sections the teachers rated important had a much higher probability of being included in the test than sections rated unimportant. For students, a similar but lower degree of correspondence between ratings and test content was found. Interestingly, for sections that teachers considered important, students more often gave a higher rating when these sections appeared in the test than when they did not. The same holds for sections that teachers considered unimportant. CONCLUSIONS: For both teachers and students there is a limited correspondence between perceived task demands and test content. Furthermore, students' perceptions of task demands show a compensation for some of the differences between their teachers' intended task demands and the test demands. PMID- 15130189 TI - Material appropriate processing and elaboration: The impact of balanced and complementary types of processing on learning concepts from text. AB - BACKGROUND: The Material Appropriate Processing (MAP) framework suggests that the influence of a text adjunct on the learning and transfer of textual information will be moderated by the overlap between type of processing induced by the adjunct and by the organisation of the text. The two types of processing are item specific processing and relational processing. Although complementary types of processing have been found to produce superior concept learning effects in previous research, there is some question as to the effects of complementary but unbalanced processing. AIMS: This study examined the effects of different combinations of two types of text adjuncts (i.e., elaborative activities) and two types of text on learning concepts from text. The four combined treatments differed as to the degree to which they were balanced and/or complementary. SAMPLE: Participants were 80 undergraduate students who were enrolled in a Year 3 education paper. METHODS: Students studied a passage that included adjuncts which asked them to either: (a) create personal examples of target concepts, or (b) contrast the target concepts. In addition, two versions of text were paired with these adjuncts: specific-only text and specific/relational text. Subjects took a criterion test that consisted of cued recall of definitions, free recall of text, classification of novel examples, and problem solving. RESULTS: Best performance occurred within the condition that included balanced and complementary processing of text/adjunct information, and worst performance occurred in the condition that included non-complementary processing. CONCLUSION: Although these results are consistent with a MAP perspective, the results are equivocal about the potential interfering effects of complementary and unbalanced processing on learning of concepts from text. PMID- 15130190 TI - Re-examining the effects of noncontingent success on self-handicapping behaviour. AB - BACKGROUND: Self-handicapping refers to the practice on the part of certain individuals to handicap their performance when poor performance is likely to reveal low ability. Noncontingent success (feedback that is inflated relative to performance) is more likely to promote self-handicapping behaviour than noncontingent failure (failure feedback based on false or misleading information). However, the reasons for the differing effects of these forms of performance feedback on self-handicapping behaviour remain obscure. AIMS: The present study sought an explanation for the differing effects of these forms of performance feedback, testing the assumption that students high in self handicapping behaviour would react more negatively following noncontingent success, reporting more unstable and external attributions, higher anxiety, and a greater propensity to claim handicaps than those low in self-handicapping behaviour. No differences were expected on any of these measures for high relative to low self-handicappers following either noncontingent failure or success. SAMPLE: Participants were 72 undergraduate students, divided equally between high and low self-handicapping groups. METHOD: High and low self handicappers were assigned to one of three performance feedback conditions: noncontingent failure, success and noncontingent success. High and low self handicappers were then given an opportunity to claim handicaps prior to completing measures of attributions and state anxiety. Subsequently, they completed 12 remote associate tasks, serving as an assessment of performance, and 16 unicursal tasks, assessing practice effort. RESULTS: Following noncontingent success, high self-handicappers reported greater anxiety, more unproductive attributions and claimed more handicaps than low self-handicappers. However no differences were evident for high and low self-handicappers following either noncontingent failure or success. High self-handicappers also performed poorly on the remote associates tasks and reduced practice effort on the unicursal tasks. CONCLUSIONS: These findings confirm the adverse effects of noncontingent success for high self-handicappers, while failing to provide evidence that noncontingent failure has any more adverse effects on high relative to low self-handicappers. PMID- 15130191 TI - Examining the multidimensionality of approaches to learning through the development of a revised version of the Learning Process Questionnaire. AB - AIM: This study aimed to produce a revised two-factor version of the Learning Process Questionnaire (R-LPQ-2F) with deep and surface approach scales, measured by a reasonably small number of items, suitable for use by teachers in secondary schools to evaluate the learning approaches of their students. METHOD: A set of 41 items was derived, with modification, from the original version of the LPQ and from items used to develop the revised version of the SPQ. These items were tested using reliability procedures and confirmatory factor analysis and items were deleted until scales were of a suitable length and confirmatory factor analysis indicated a good fit to the intended two-factor structure. SAMPLE: The sample consisted of 841 students from 20 secondary schools in Hong Kong. RESULTS: The final two-factor version of the questionnaire had good Cronbach alpha values and reasonable goodness-of-fit values for the confirmatory analysis. There was a much better fit, though, to a hierarchical structure with motive and strategy subscales for each approach, each of which, in turn, had two subcomponents. CONCLUSION: Approaches to learning have a hierarchical dimensionality with motive and strategy elements. Each motive and strategy element is itself multidimensional. The results are used to question the conventional approach to the testing and acceptance of instruments, which place sole reliance upon reliability tests. The use of confirmatory factor analysis is recommended as a routine procedure in the development and testing of instruments. PMID- 15130192 TI - The effects of competition on achievement motivation in Chinese classrooms. AB - BACKGROUND: Laboratory studies have consistently found that competition induces performance goals and affects learning motivation. However, the ecological validity of these results is yet to be established. There is a need for investigation of whether the results hold in both the classroom context and non Western culture. AIM: The study investigated the effects of competition on learning motivation among Chinese students in an authentic classroom setting. SAMPLE: The participants were 52 students of grade 7 from two Hong Kong secondary schools. METHOD: They were randomly assigned to either competitive or non competitive conditions in a 2-hour Chinese typewriting course. RESULTS: Students in the competitive condition performed better in easy tasks than their counterparts in the non-competitive condition. However, they were more performance-oriented and more likely to sacrifice learning opportunities for better performance. They were also prone to have worse self-evaluation after failure. Although there were no statistically significant differences between the two conditions in task enjoyment and achievement attribution, the direction of the differences was consistently unfavourable to students in the competitive condition. CONCLUSION: The findings were consistent with the predictions of goal theory. Competitiveness induces performance goals and worse self-evaluation after failure among Chinese students in a classroom setting, as was found with Western students in a laboratory setting. PMID- 15130193 TI - Bully/victim problems and their association with Machiavellianism and self efficacy in Greek primary school children. AB - BACKGROUND: Previous research demonstrated that Machiavellian beliefs are linked with bully/victim problems at school. However, Machiavellianism was treated as a single construct and not as multidimensional. Children's perceptions of self efficacy in both social and academic domains have been related to conflictual peer interactions but not directly to bully/victim problems. This study extends previous work by examining the association of Machiavellianism and self-efficacy with bully/victim problems. AIMS: The main purpose of this study was to examine the relationship between bully/victim problems and (a) components of Machiavellian beliefs, (b) Self-efficacy for Assertion, (c) Self-efficacy for Aggression and (d) Self-efficacy for Learning and Performance, among school-age children. It was also examined whether children who bully others and are bullied themselves (bully/victims) are a distinct group in terms of Machiavellian beliefs and the above perceptions of self-efficacy. SAMPLE: The sample consisted of 186 children drawn from the fourth to sixth grade classrooms of four primary schools in central Greece. METHOD: Peer victimization and bullying behaviour were assessed by two 6-item self-report scales (Austin & Joseph, 1996), Machiavellian beliefs with a 20-item scale (Christie & Geis, 1970), Self-efficacy for Assertion and Self-efficacy for Aggression with two 6-item scales (Egan & Perry, 1998) and Self-efficacy for Learning and Performance with an 8-item scale (Pintrich, Smith, Garcia, & McKeachie, 1991). Higher scores reflected greater victimization, bullying behaviour, Machiavellianism and domain specific self-efficacy. Data were analysed using both correlational and categorical approach. RESULTS: Factor analysis of the Kiddie Mach scale revealed four main factors: Lack of Faith in Human Nature, Manipulation, Dishonesty and Distrust. The results of the correlational approach suggest that both bullying and victimization are associated with most of these factors, overall Machiavellianism and self-efficacy measures. Separate analyses for boys and girls provide a more precise picture of that association. The results of the categorical approach, regarding differences in the Kiddie Mach and Self-efficacy scales between bullies, victims and bully/victims, suggest that bully/victims are a distinct group in terms of Lack of Faith in Human Nature and overall Machiavellianism. Moreover, bully/victims were found to be similar to victims with respect to Self-efficacy for Assertion but similar to bullies with respect to Self-efficacy for Aggression. CONCLUSIONS: Anti-bullying interventions might profit from a greater focus on mastery-oriented motivation and more emphasis on citizenship and interpersonal relationships in order to minimize children's aggression-encouraging cognition and reduce Machiavellian attitudes. Further research is needed to explain satisfactorily the behaviour patterns of bully/victims. PMID- 15130198 TI - Reflections: brain failure revisited. PMID- 15130199 TI - Development of a medical subspecialty in palliative medicine: progress report. AB - There is significant interest in seeking professional recognition of expertise in caring for people with serious life-threatening illness and their families through creation of a specialty in palliative medicine. Certification of physicians and accreditation of training programs are key elements for formal recognition. The American Board of Hospice and Palliative Medicine was established to achieve these goals. The next step in the maturation of the subspecialty of palliative medicine is to have both the certification and the accreditation recognized by the professional self-governing bodies in organized medicine. This paper answers common questions about obtaining recognition by the Accreditation Council of Graduate Medical Education, the American Board of Medical Specialties and its member boards. Formal recognition of the subspecialty of palliative medicine is sought in order to extend the knowledge and skills inherent in the domains of palliative medicine. Such recognition will also encourage more physicians to enter the field and assure standards of care for those patients and their families who need it. PMID- 15130200 TI - End-of-life care in nursing homes: residents in hospice compared to other end stage residents. AB - OBJECTIVE: To compare residents in hospice care at admission to the nursing facility to end stage residents not in hospice at admission. DESIGN AND METHODS: We analyzed 18,211 admission assessments recorded in the Minimum Data Set (MDS) during the year 2000 throughout the United States for residents classified as having an end-stage disease (6 or fewer months to live). Fifty-nine percent (n = 10,656) of these residents were in hospice care at the time of their admission assessment. We used these MDS admission assessments to compare residents in hospice care to other end-stage residents not in hospice for demographic characteristics, health status, and treatments. RESULTS: Hospice residents at admission were significantly more likely to be female, older, white, and widowed than other end-stage residents at admission. There were significant differences between hospice residents and other residents at end stage in the use of advanced directives at admission. Hospice residents at admission experienced significantly more frequent and more intense pain than other end-stage residents at admission, while these hospice residents also showed greater impairment in cognitive ability and physical function. While cancer was the most common disease among these end stage residents, it was significantly more prevalent among hospice residents. IMPLICATIONS: Many end-stage residents may not be receiving adequate palliative care in nursing facilities; further study of this is warranted. The MDS should be revised to record minimum standards for palliative care with or without the use of hospice to improve end-of life care in nursing facilities. PMID- 15130201 TI - Measuring patient treatment preferences in end-of-life care research: applications for advance care planning interventions and response shift research. AB - Understanding the dynamics of patient treatment preferences can be important for end-of life are research, and has particular salience not only to guide a process of advance care planning (ACP) but also as an outcome measure. Ascertaining the reliability and responsiveness of preferences for life-sustaining treatments within and between patients is a necessary foundation for utilizing patient-agent congruence as an outcome for ACP interventions. This study validated a modified version of the Emanuel and Emanuel Medical Directive for use in both research and clinical applications. Seriously ill patients (n = 168) were asked at baseline and 21 days to consider four common end-of-life health state scenarios, to indicate their goals for treatment, and to state their preferences for six specific treatments. We investigated the reliability and validity of this tool. We found that preferences for life-sustaining treatments were highly intercorrelated, and internally consistent across treatments by scenario and across scenarios by treatment. Preferences for pain medications were, however, distinct from preferences for other treatments. Preference scores exhibited stability over follow-up, and demonstrated both concurrent and discriminant validity. We detected a small effect size for change in preferences as a function of health state change, suggesting that re-prioritization response shifts do occur but are small in magnitude in these patient samples over this time frame. We conclude that this measure is reliable and valid for use in clinical settings and for evaluating interventions designed to improve patient-agent congruence about patient preferences for life-sustaining treatments. Clinical applications of the tool are discussed. PMID- 15130202 TI - Knowledge, attitudes, and beliefs about end-of-life care among inner-city African Americans and Latinos. AB - OBJECTIVE: This project explored end-of-life care preferences and barriers among low-income, urban African Americans and Latino/Hispanic Americans (Latinos) to uncover factors that may influence hospice utilization. METHODS: Focus groups were conducted separately for African Americans (4 groups, n = 26) and Latinos (4 groups, n = 27). Transcripts were coded and analyzed using consensus and triangulation to identify primary themes. RESULTS: Four preference themes and four barriers were identified. Results were largely similar across the two groups. Both preferred having families provide care for loved ones but expressed desire to reduce caretaker burden. Groups emphasized spirituality as the primary means of coping and valued the holistic well-being of the patient and family. Barriers reported were closely tied to access to care. Participants reported low hospice utilization because of lack of awareness of hospice and the prohibitive cost of health care. Latinos were more likely to report language barriers, while African Americans were more likely to report mistrust of the system. CONCLUSIONS: African Americans and Latinos in this study were highly receptive to end-of-life care that would provide relief for patients and caregivers and emphasize spirituality and family consensus. Improving awareness of hospice services would likely increase utilization. PMID- 15130203 TI - Hydration management at the end of life. AB - The management of parenteral hydration at the end of life remains controversial. The debate centers on whether and/or how often patients should be hydrated, the volume of hydration received, and the benefit verses side effects of parenteral hydration. In order to clarify the routine practice of physicians involved in the end-of-life care in Edmonton, Alberta, Canada, we investigated the routine management of hydration by attending physicians caring for patients dying in a palliative care unit (PCU) at Norwood Capital Care, and in acute care wards at the Royal Alexandra Hospital (RAH) both while receiving and while not receiving consult advice from the Palliative Care Program. We conducted a retrospective chart review of 50 consecutive patients who died in each of the 3 sites included in the study. Data from the last 7 days prior to and including the date of death (day 0) was recorded. The majority of patients at all sites received hydration. The volume of hydration ordered in the Norwood PCU site was significantly different compared to both RAH groups on all days studied (p < 0.005). The RAH palliative care group showed a trend for lower hydration volumes compared to the RAH acute care group with significant differences on days 1 and 2 (p < 0.05). Throughout the week, for all of the hydrated patients in the Norwood PCU site, hypodermoclysis (HDC) was ordered; for nearly all of the hydrated patients in the RAH acute care group, intravenous (IV) hydration was ordered; and for approximately one third of the hydrated RAH palliative care consult group HDC was ordered, and for the remainder IV hydration was ordered. The RAH acute care group represented the largest percentage of hydrated patients receiving diuretics while the Norwood hospice site represented the lowest. The data raise the possibility that more patients in the RAH acute care group were overhydrated and may have developed symptoms such as edema, ascites, and respiratory distress. This study suggests that hydration at the end of life is managed differently in different settings of care and highlights areas for education to improve management. PMID- 15130204 TI - Decompressive tube esophagostomy: a forgotten palliative procedure? AB - Many patients with complete, irreversible upper gastrointestinal (GI) tract obstruction will require decompression for relief of intractable nausea and vomiting. Nasogastric (NG) tubes are associated with patient discomfort and risk. Gastrostomy tubes may not be technically feasible in a small subset of patients with advanced upper GI tract malignancy. Decompressive tube esophagostomy is an underutilized, minimally invasive alternative in such patients. We present a case report, a description of the procedure, and a review of the literature for this palliative procedure. PMID- 15130205 TI - Where infants die: examination of place of death and hospice/home health care options in the state of Wisconsin. AB - Infants (less than 1 year of age) have the highest death rates in the pediatric population, yet there is little published on hospice utilization for infant home deaths. We sought to describe: (1) where infants with a predisposing life threatening condition are dying, (2) agency services available to dying infants and their families, and (3) utilization of these services for infants within the state of Wisconsin. We collected information from death certificates for infants whose cause of death was either congenital anomaly or condition of the perinatal period, such as hypoxic ischemic encephalopathy or prematurity. In addition, we surveyed all hospice and home health agencies in Wisconsin to determine their ability to serve and whether they were utilized for this same population. During 1992-1996 in Wisconsin, state records indicate that 2591 infants died: congenital anomalies or conditions of the perinatal period resulted in 1538 (60%) of these deaths. Of the 508 infant deaths from congenital anomalies, 46 (9%) occurred at home. Of the 1030 deaths from conditions of the perinatal period, 16 (1.5%) occurred at home. Only 36 (40%) of the 91 hospice/home health agencies that responded to our survey provided services to the pediatric population between 1992-1996. During this time, only 11 agencies provided care for 20 infant home deaths, comprising 32% of infant home deaths reported to the state in that same time period. In comparison to adults and older children, we found a low home death rate for infants with a life-threatening condition. To clarify these findings, we discuss barriers to infant home death. PMID- 15130206 TI - Palliative educational outcome with implementation of PEACE tool integrated clinical pathway. AB - BACKGROUND: House officers frequently lack basic competency in end-of-life care. Few studies have evaluated educational interventions deliberately utilizing physicians' learning strategies, particularly in the context of a concomitant effort at modification of practice patterns. STUDY DESIGN: Prospective controlled trial utilizing pre-intervention and post-intervention cross-sectional surveys. PARTICIPANTS: Internal medicine residents at a university hospital in their first, second, and third years of training. SURVEY: A 25-item survey modified from previously published instruments. INTERVENTION: Residents in the intervention group utilized an experiential learning intervention (integrated, end-of-life clinical pathway: PEACE Tool). The control group delivered care in a standard fashion. DATA ANALYSIS: SURVEY item and test responses were tabulated and pair-wise comparisons between group means evaluated statistically using two sample t tests. RESULTS: Fifty-four internal medicine residents (n = 24, first year; n = 17, second-year; and n = 13, third-year) completed the survey. Pre intervention mean scores on a 16-item knowledge scale were 7.4 (46% correct) for first-year, 8.1 (51%) for second-year, and 9.2 (58%) for third-year residents. Eighteen first-year residents participated in the intervention phase (8 in the intervention, 10 in the control). Mean overall knowledge scores were 46% higher in the intervention group compared to the control group (11.8 versus 8.1 p < 0.001). CONCLUSIONS: A time-effective, practice-based strategy led to a significant improvement in knowledge of end-of-life care. Prior to implementation of this strategy competency in end-of-life care was suboptimal among internal medicine residents, in spite of desirable attitudes. Factual knowledge improved slightly with standard, pre-intervention training and experience. PMID- 15130207 TI - The spirit feather: an ecologically based celebration of life. AB - The final moments of a life pose a special and very important time for all participants. Sensitive arrangement of events can provide harmony and a peaceful and memorable transition event. A specific situation, in which a feather is used to symbolize the transition, is reported. Hospice programs can offer ecologically based celebration of life and provide a positive transition for patients, families, friends, and staff. PMID- 15130208 TI - Unequal access to hospice and palliative care. PMID- 15130209 TI - Methadone for pain: No. 75. PMID- 15130210 TI - Methadone: starting dosing information No. 86. PMID- 15130212 TI - Normal grief and bereavement: letters from home. PMID- 15130214 TI - Communication, trust, and making choices: advance care planning four years on. PMID- 15130215 TI - Shifting the focus of advance care planning: using an in-depth interview to build and strengthen relationships. PMID- 15130216 TI - Informing the patient-proxy covenant: an educational approach for advance care planning. PMID- 15130217 TI - Adapting advance medical planning for the nursing home. PMID- 15130218 TI - Project ENABLE: a palliative care demonstration project for advanced cancer patients in three settings. AB - At the end of the 1990s, based on data from two major studies of end-of-life (EOL) care, the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatment (SUPPORT), and the Hospitalized Elderly Longitudinal Project (HELP), a consensus panel report documented the problems and needs of patients with cancer and other life-limiting diagnoses at end-of-life. A national program of The Robert Wood Johnson Foundation (RWJF), Promoting Excellence in End-of-Life Care, attempted to address these needs by funding demonstration projects to test various approaches to improve identified deficits. In 1998, Project ENABLE (Educate, Nurture, Advise Before Life Ends), one of four RWJF-funded cancer center/hospice collaborations of the Promoting Excellence program, began to address these issues. The jointly sponsored Norris Cotton Cancer Center (NCCC)/Hospice of Vermont and New Hampshire (Hospice VNH) program provided an integrated approach to the management of life-limiting cancer. Project ENABLE was aimed at alleviating the symptoms of disease and treatment, enhancing clinician and patient/family communication, offering support for families, friends and other caregivers, addressing emotional and spiritual needs of dying people and providing conceptual and administrative structure to provide EOL care consistent with patients' values and preferences. Although patient symptom data is not yet available, other measures of success included improved access to hospice and palliative care services from the time of diagnosis and a sustained palliative care program at two of the three sites in which the program was implemented. PMID- 15130219 TI - The GRACE Project: Guiding End-of-Life Care in Corrections 1998-2001. AB - The GRACE (Guiding Responsive Action in Corrections at End of Life) Project promotes the development of end-of-life (EOL) care programs in correctional settings. Managers of pioneer programs were interviewed and worked together with correctional and community professionals in the planning. The project established a resource center, developed and published standards and a handbook, and produced journal articles. Four sites with pioneering programs have undertaken demonstration projects to show how program enhancements could be designed, implemented, monitored, and evaluated. Early data suggest that both frontline staff and supporting organizations have crucial roles to play in sustaining present momentum. When representatives from diverse areas of expertise worked together on this project, innovations emerged that help point the way for future institutional change. PMID- 15130222 TI - Leptin in morbidly obese patients: no role for treatment of morbid obesity but important in the postoperative immune response. AB - Despite the current opinion that leptin can no longer be seen as a hormone which could be used therapeutically to prevent an energy surplus (it rather protects the organism for an energy deficit), leptin may still have an impact in clinical medicine. Leptin was shown to have several important functions. The pleiotropic properties of leptin include a regulatory function in the immune system. Reviewing the effects of leptin on different parts of the immune system reveals that the immune system is deregulated in an environment low in leptin. A strong reduction in leptin levels occurs in situations of starvation as seen after bariatric surgery. We postulate the hypothesis that the starvation-induced postoperative decrease of leptin is causative of the more serious course of complications observed after bariatric surgery. PMID- 15130223 TI - Stenosis of the gastroenterostomy after laparoscopic gastric bypass. AB - BACKGROUND: Stenosis of the gastroenterostomy after laparoscopic Roux-en-Y gastric bypass (LRYGBP) is a serious problem that occurs after stapled (linear or circular) and hand-sewn anastomoses. METHODS: Data was prospectively entered into a database to track complications of bariatric surgery. Between Feb 27, 1999 and June 13, 2000, 1000 patients underwent LRYGBP. All patients met NIH criteria for bariatric surgery.The gastroenterostomy was constructed with a linear stapler inserted to 20 mm (15 mm cut). The stapler defect was closed with a polyester running suture to construct a 12-mm diameter anastomosis. The anastomosis was banded with fascia lata to prevent late enlargement. All patients with suspected stenosis were endoscoped. RESULTS: 32 patients (3.2%) developed stenosis (<10 mm diameter orifice) at the gastroenterostomy. There were 27 females. Average age was 44.8. Average BMI was 45.0. Average stenotic orifice was 5.7 mm in diameter. Stenoses occurred in 18 of 32 patients (56.3%) by 3 months, 26 (81.3%) by 6 months, and 31 (96.9%) by 1 year. 30 of the 32 patients underwent endoscopic dilation as initial therapy. 17 of the 32 underwent multiple dilations. Dilation caused 4 perforations, resulting in 2 emergency operations. Perforation occurred at the first attempt at dilation in 3 of 4 patients. Stenoses could not be successfully dilated in 8 patients, and all 8 underwent surgical revision. There were no deaths in these 32 patients, but there were 68 dilations and 10 re operations. CONCLUSION: Stenosis of the gastroenterostomy after LRYGBP is an infrequent but serious problem, which results in considerable morbidity. PMID- 15130224 TI - Longitudinal gastrectomy as a treatment for the high-risk super-obese patient. AB - BACKGROUND: Bariatric surgery in patients with significant co-morbid conditions is associated with increased perioperative risk. METHODS: From 1995-2001, 795 patients were operated upon at our institution for the diagnosis of morbid obesity. Of these, 671 (84.4%) had the duodenal switch (DS) procedure. Longitudinal gastrectomy (LG) entails a greater curvature linear gastrectomy creating a gastric tube with a volume of 100 ml along the lesser curvature of the stomach. This procedure was performed for 21 patients (median age 50.5, median BMI 56). 9 patients were offered LG preoperatively because of their known high perioperative risks. 12 patients were initially planned for DS, but the procedure was limited to LG alone because of either unexpected intraoperative findings (n=9) or intraoperative hemodynamic instability (n=3). 5 patients developed complications, and there were no deaths. RESULTS: 19 out of 21 patients were available for a median follow-up of 17.5 months (6.25-20.25). Median weight loss and median %EWL at 12 months were 44.5 kg and 45.1%, respectively. Estimated daily dietary volume at 1 year was 35% of preoperative values. Of 10 patients followed for >or= 1 year, 4 of 10 achieved more than 50% EWL and 8 patients were taking less or were completely off medications for diabetes, hypertension and congestive heart failure. Weight loss plateaued at 1 year for the majority of patients. CONCLUSIONS: LG is a safe and effective option for high-risk morbidly obese patients. Weight reduction is accomplished by limitation of caloric intake. LG can be offered to high-risk morbidly obese patients as an interim procedure to help decrease perioperative risk before DS. PMID- 15130225 TI - Post-anesthesia recovery after infusion of propofol with remifentanil or alfentanil or fentanyl in morbidly obese patients. AB - BACKGROUND: The type of opioid used during general anesthesia in the morbidly obese influences recovery and the postoperative period. In a randomized clinical trial, the postoperative recovery profile and early period after general anesthesia with remifentanil, fentanyl and alfentanil were compared in morbidly obese patients. MATERIAL AND METHOD: 60 morbidly obese patients with BMI >35 kg/m(2) (mean 43.31) undergoing open Roux-en-y gastric bypass were randomly divided into 3 groups: remifentanil (R), fentanyl (F), and alfentanil (A). Dosage of opioids was based on ideal body weight (IBW): fentanyl 5 mcg/kg for intubation followed by infusion of 0.025-0.05 mcg/kg/min; alfentanil 15 mcg/kg initially, then 1.0-1.5 mcg kg/min; and remifentanil 1 mcg/kg followed by infusion of 0.25 1.5 mcg/kg/min. Anesthesia was induced with infusion of propofol and oxygen with N(2)O (1:1). After anesthesia, the duration to response to verbal command, spontaneous respiration, adequate respiration, and safe extubation were recorded. The incidence of postoperative nausea and vomiting were recorded. Using verbal scale for evaluation of postoperative pain, the early postoperative analgesia requirements were assessed. RESULTS: Demographic profiles and duration of procedure did not differ between groups. A total dose of propofol was significantly lower in Group R compared with Groups A and F (P <0.05). Duration to spontaneous respiration, adequate respiration and safe extubation were significantly shorter in Group R compared with Group F (P <0.05). Shortly after anesthesia, significantly more patients in Group R required additional dose of analgesic than in Group F (P <0.05). Postoperative nausea and vomiting (PONV) occurred significantly more often in Group R compared with Group F (P <0.05). Recovery profile of Group A was more similar to Group R, and postoperative pain and PONV evaluation more similar to Group F. CONCLUSION: In morbidly obese individuals, alfentanil or fentanyl and remifentanil can be safely used, but there is a higher rate of PONV and postoperative pain in the remifentanil group. PMID- 15130227 TI - Elevated liver enzymes following open Roux-en-Y gastric bypass for morbid obesity - does timing of liver retraction affect the rise in the levels of transaminases? AB - BACKGROUND: Liver retraction in open Roux-en-Y gastric bypass (RYGBP) for morbid obesity may cause an elevation in liver transaminase levels postoperatively. This study attempts to ascertain whether timing of placement of the liver retractor would have any effect on the degree of derangement of the liver function tests. METHODS: 12 patients undergoing open RYGBP were prospectively randomized into 2 groups. One group had the liver retractor applied early on in the operation, and had the gastric pouch constructed first, followed by construction of the jejunojejunostomy and Roux-en-Y gastrojejunostomy. The second group had the jejunojejunostomy performed first, followed by placement of the liver retractor to allow construction of the gastric pouch and Roux-en-Y gastro-jejunostomy. Liver function tests were monitored postoperatively in both groups, and these were then statistically analyzed with respect to the duration of liver retraction. RESULTS: Delayed liver retraction resulted in statistically significantly smaller increases in aspartate transaminase (AST) and alanine transaminase (ALT) compared with early liver retraction. AST levels returned to normal values within 1 week in both groups of patients. ALT levels returned to normal values within 1 week in the delayed liver retraction group, whereas levels took >1 week to return to normal in the early liver retraction group. CONCLUSIONS: Liver retraction causes abnormal transaminase levels following open RYGBP. Delayed liver retraction results in less rise in the AST and ALT levels compared with early liver retraction. Surgeons should construct the Roux-en-Y limb and jejunoje junostomy first, before proceeding with construction of the gastric pouch and gastrojejunostomy, thereby decreasing the duration of liver retraction and consequent relative liver ischemia. PMID- 15130228 TI - Improvement of hypothyroidism after laparoscopic Roux-en-Y gastric bypass for morbid obesity. AB - BACKGROUND: Laparoscopic Roux-en-Y gastric bypass (LRYGBP) has been very effective in managing a broad range of morbid obesity-related co-morbidities. We report a beneficial effect of LRYGBP that has not been previously observed. METHODS: Between December 1999 and September 2002, 224 patients underwent LRYGBP. Preoperative assessment for hypothyroidism and follow-up data were prospectively collected in our database. Improved thyroid function (ITF) or unchanged thyroid function (UTF) was determined by comparison of preoperative and postoperative thyroxine requirements. RESULTS: 23 of 224 patients (10.3%) were treated preoperatively for hypothyroidism. During a median follow-up of 17 months, hypothyroidism was improved in 10/23 patients (43.5%). 2 patients had complete resolution, and the remaining 8 had reduction (14%-50%) of their thyroxine requirements. ITF occurred at a mean follow-up of 8.9 months and at a mean excess weight loss (EWL) of 57%. 6 of the 8 patients (75%) with ITF >or= 25% had EWL >90% at last follow-up, compared to 1 out of 15 patients (6.6%) with UTF or <25% improvement (P =0.001). Comparison of patients with ITF and UTF over time during a 20-month follow-up, showed no significant difference in mean body mass index (BMI) and mean percentage of EWL. CONCLUSION: Improvement of hypothyroidism may be an additional benefit of bariatric surgery that has not been previously reported. Reduction of thyroxine requirements is most likely the result of the decrease in the BMI. PMID- 15130229 TI - Impact of patient follow-up on weight loss after bariatric surgery. AB - BACKGROUND: Postoperative follow-up after bariatric surgery is important. Because of the need for adjustments, follow-up after gastric banding may have a greater impact on weight loss than after Roux-en-Y gastric bypass. We reviewed all patients at 1 year after these two operations. METHODS: During the first year after surgery, laparoscopic adjustable gastric banding (LAGB) patients were followed every 4 weeks and Roux-en-Y gastric bypass (RYGBP) patients were followed at 3 weeks postoperatively and then every 3 months. The number of follow up visits for each patient was calculated, and 50% compliance for follow-up and weight loss was compared. RESULTS: Between October 2000 and September 2002, 216 LAGB and 139 RYGBP operations were performed. Of these patients, 186 LAGB patients and 115 RYGBP patients were available for 1-year follow-up. Age and BMI were similar for each group. Overall excess weight loss (EWL) after LAGB was 44.5%. 130 (70%) returned 6 or less times in the first year and achieved 42% EWL. 56 patients (30%) returned more than 6 times and had 50% EWL (P=0.005). Overall %EWL after RYGBP was 66.1%. 53 patients (46%) returned 3 or less times in the first year, achieving 66.1% EWL. 62 patients (54%) returned more than 3 times after surgery and achieved 67.6% EWL (P=NS). CONCLUSION: Patient follow-up plays a significant role in the amount of weight lost after LAGB, but not after RYGBP. Patient motivation and surgeon commitment for long-term follow-up is critical for successful weight loss after LAGB surgery. PMID- 15130230 TI - Dynamic radioisotope scintigraphy for gastric banding adjustment. AB - BACKGROUND: Gastric banding is today a common restrictive procedure to treat morbid obesity. After operation, adjustment of the band is required to adequately restrict food intake, allowing better results. The aim of this study was to evaluate the results of a unique method of adjustment using dynamic radioisotope scintigraphy. METHODS: 40 patients after Laparoscopic Adjustable Gastric Banding (LAGB) were prospectively divided into 2 equal groups, and the results of adjustment analyzed using the conventional method of fluoroscopic image and barium swallow (FA Group), and our proposed new method of dynamic radioisotope scintigraphy (DRS Group) using Tc99-phytate labeled plain yogurt. RESULTS: After 6 months follow-up, the FA group lost 12.34%, while the DRS group lost 20.34% of their initial weight. In the FA group, we found that 95% of the patients needed re-adjustments versus only 25% in the DRS group (P <0.0001). Vomiting was more frequent in the FA group than in the DRS group, mean 0.65 and 0.2, respectively (P < 0.009). The radiation exposure was 4 times higher in the FA group, but the cost of the 2 adjustment procedures was similar. CONCLUSION: DRS is a more physiologically friendly and accurate method of gastric banding adjustment than the conventional barium swallow adjustment; it also results in a better weight loss during the first 6 months after operation. PMID- 15130231 TI - Mechanical resistance of gastric bands - comparison between new bands and bands within the patient for at least two years. AB - BACKGROUND: Morbid obesity is a rapidly increasing health risk in industrialized countries, and is associated with serious co-morbidities. Since conservative medical therapies fail to sustain significant weight loss, adjustable gastric banding (AGB) has become an established therapy for morbid obesity. To our knowledge there have been no trials assessing whether gastric bands implanted in the patient for some time can withstand the same mechanical stress as a new band. METHODS: The mechanical resistance of unused and used gastric bands was tested through the Tensile Test, to evaluate if a material is strong and rigid enough to withstand the loads experienced in use. 9 bands were tested, 2 of which were unused. RESULTS: The tested new Swedish AGB (SAGB) resisted 361 Newtons (N, 36.8 kg) until tearing, and the new LAGB (Inamed) 157 N (16 kg). The 7 SAGB which had been in a patient for at least 2 years, pulled apart at a mechanical stress of 250.6 N (25.54 kg). CONCLUSION: The new SAGB had a higher mechanical tensile strength than any of the used bands placed in the body for 2 years. PMID- 15130232 TI - MMPI-2 scores in the outcome prediction of gastric bypass surgery. AB - BACKGROUND: A psychological assessment is critical for morbidly obese patients seeking Roux-en-Y gastric bypass (RYGBP) surgery. The Minnesota Multiphasic Personality Inventory (MMPI) has been widely used in past psychological studies of bariatric surgery patients, but, to date, there is no published research on the more recent version of the MMPI, the Minnesota Multiphasic Personality Inventory-2 (MMPI2), and its relation to RYGBP outcome. This investigation was designed to evaluate the predictive validity of the MMPI-2 with respect to outcome of RYGBP for morbid obesity. METHODS: The research involved a retrospective analysis of MMPI-2 scores of 2 groups of patients 1 year following RYGBP: 1). those who lost >or= 50% of their excess weight and 2). those who lost <50% of their excess weight. Subjects were 52 morbidly obese patients (mean age 44 years, mean BMI 56 kg/m2). The measurement of psychological variables consisted of the MMPI-2 scores of 3 validity scales, 10 clinical scales, and 3 Content Scales, and BMI. RESULTS: Those who lost <50% excess weight scored significantly higher than those who lost >50% excess weight on the F,Hysteria, Paranoia, and Health Concerns scales of the MMPI-2, and significantly lower on the Masculinity-femininity scale. Stepwise regression analysis found that a combination of the Health Concerns and Masculinity-femininity scales was the most accurate predictor model for 1-year post-surgery weight loss. CONCLUSION: A standard personality measure, the MMPI-2, appears to be associated with weight loss outcome 1 year after RYGBP. Psychological traits such as anxiety and excessive health concerns are likely to influence bariatric surgical outcome. PMID- 15130233 TI - Class III obesity continues to rise in African-American women. AB - BACKGROUND: The Behavior Risk Factor Surveillance System (BRFSS) survey, a random telephone survey of the United States, found a rising prevalence of Class III obesity, body mass index (BMI) >40 kg/m(2). African-American women were largely responsible for this increase. The prevalence of Class III obesity in African American women in the BRFSS during the year 2000 was 6%, identical to the Pennington Biomedical Research Center (PBRC) database during the same year. The PBRC Database might predict the trend in class III obesity in the years 2000 2002. METHODS: The PBRC is an academic research center in Baton Rouge, that maintains a database with information collected by telephone regarding BMI and race, for subjects responding to solicitations to participate in clinical trials. This database contains 7787 subjects entered from 1995 through 2002, of which 1820 were African-American women and 3006 were Caucasian women. The prevalence of Class III obesity was plotted for women of these 2 racial groups and compared with the year 2000 prevalence in the BRFSS. RESULTS: The prevalence of Class III obesity in African-American women increased from 0.5% in 1995 to 6% in 2000 (identical to the BRFSS) and to 7.5% in 2002. The prevalence of Class III obesity in Caucasian women remained about 3%. CONCLUSION: The prevalence of Class III obesity continues to rise in African-American women, a group with greater need and less access to obesity surgery, the only effective treatment, than Caucasian women, an inequality deserving of public policy attention. PMID- 15130234 TI - Vertical banded gastroplasty: is it a durable operation for morbid obesity? AB - BACKGROUND: Long-term follow-up is important in the evaluation of results of bariatric operations. The authors followed vertical banded gastroplasty (VBG) in an ongoing database. METHODS: From 1986 to 1999, 123 VBGs were performed in one surgical ward. The series comprises 105 females and 18 males, with mean age 37 years and mean preoperative BMI 48.39 kg/m(2). The authors evaluated long-term results (patients followed, patients who underwent revisional surgery, success rate and complications), analyzed in 2000 and again in 2002. RESULTS: With time, the percentage of patients in the follow-up decreased from 65 to 53%, the success rate decreased from 46.16 to 32.79%, and revisional surgery for inadequate weight loss or complications increased from 9.61 to 29.51% in only 2 years. CONCLUSION: The weight loss of VBG was frequently not maintained in the long-term. However, we have previously found maintained improvements in comorbidities. PMID- 15130235 TI - Intragastric balloon for "non-morbid" obesity: a retrospective evaluation of tolerance and efficacy. AB - BACKGROUND: One potential indication for intra-gastric balloon is weight reduction for mild to moderate obesity. The authors evaluated retrospectively the tolerance and efficacy of the BioEnterics intragastric balloon (BIB). METHODS: From February 1998 to July 2001, an intragastric balloon was placed under endoscopic control in 176 patients (mean BMI 31 kg/m(2)). It was filled with 500 ml saline in the first 142 patients and with 600 ml in the last 34. Removal was proposed between 4 and 6 months after balloon insertion. RESULTS: Balloon placement was uneventful. 13 patients were lost of follow-up (7.4%). Removal was performed endoscopically in 113 patients (64.2%), with 1 case of tracheal aspiration. Balloon evacuation was spontaneous in 49 cases. 1 BIB was removed at laparoscopic surgery for small bowel obstruction. Side-effects were: vomiting during the first week (90%), occasional vomiting for >3 weeks (18%), hypokalemia (8.5%), functional renal failure (1.1%), abdominal pain (12.5%), gastro esophageal reflux (11.5%). There were 2 gastric ulcers, 1 sub-occlusion treated endoscopically and the 1 small bowel obstruction treated surgically, occurring after the theoretical date of removal in all cases. Mean excess weight loss was 38 +/- 28.5 % (35.4 +/- 27.3 % for 500-ml balloons and 48.8 +/- 31.0 % for 600-ml balloons (P <0.02)). CONCLUSIONS: The BIB appears to be safe provided that it is removed within the period specified by the manufacturer. Its efficacy to reduce weight in patients with non-morbid obesity may depend in part on the filling volume. PMID- 15130236 TI - Implantable gastric stimulation for the treatment of severe obesity. AB - The prevalence of obesity is growing worldwide at an alarming rate. Current medical therapies are often ineffective and surgical treatments result in weight loss but have significant risk. Implantable Gastric Stimulation (IGS) offers a novel approach to weight loss. Simply stated, the IGS system electrically stimulates the stomach with a pacemaker-like device. The device is implanted in a brief minimally invasive procedure. Investigation in over 500 patients globally has proven it to be safe and seemly free of long-term sequelae. With refinements in patient selection and device application, the weight loss results have been steadily improving. The IGS may someday become a reliable and safe surgical option for weight loss. PMID- 15130237 TI - Intraluminal migration of bovine pericardial strips used to reinforce the gastric staple-line in laparoscopic bariatric surgery. AB - A 41-year-old morbidly obese woman (BMI 40.8) underwent elective laparoscopic biliopancreatic diversion with duodenal switch. The operation involved: sleeve gastrectomy, division of duodenum, creation of a duodenoenterostomy, and creation of a distal ileoenteric anastomosis. With laparoscopic stapling, bleeding is often a problem along the sleeve gastrectomy staple-line. To reduce this risk, we used bovine pericardial strips to reinforce the staple-line throughout its length. 4 weeks postoperatively, the patient was found to have fragments of pericardium in her vomitus, indicating intraluminal migration of the reinforcing strips. Her subsequent course has been uneventful. This represents the first report of such migration. Indications, benefits, complications and risks of using bovine pericardium to reinforce gastric staple-lines in laparoscopic bariatric surgery are discussed. PMID- 15130238 TI - Severe metabolic acidosis resulting from a dislocated gastric band. AB - A 32-year-old morbidly obese woman with an obstructing dislocated gastric band is presented. Because of prolonged vomiting, a metabolic alkalosis would be expected, but instead an impressive high anion gap acidosis was observed. Because of a highly positive urine ketone test and a high serum concentration of beta hydroxybutyrate, a ketoacidosis caused by starvation appears to be the primary cause of this metabolic acidosis. This type of acidosis was treated successfully with intravenous administration of glucose and insulin as well as sodium bicarbonate, with urgent removal of the band. PMID- 15130239 TI - Band erosion with gastric cancer. AB - Adjustable gastric banding is a well-established procedure for the treatment of morbid obesity. We present a 62-year-old female who experienced the rare complication of intragastric band perforation due to a gastric adenocarcinoma localized at the site of gastric banding, 10 years after insertion of the band. PMID- 15130241 TI - Advice to a patient seeking bariatric surgery. PMID- 15130244 TI - Nonlinear population codes. AB - Theoretical and experimental studies of distributed neuronal representations of sensory and behavioral variables usually assume that the tuning of the mean firing rates is the main source of information. However, recent theoretical studies have investigated the effect of cross-correlations in the trial-to-trial fluctuations of the neuronal responses on the accuracy of the representation. Assuming that only the first-order statistics of the neuronal responses are tuned to the stimulus, these studies have shown that in the presence of correlations, similar to those observed experimentally in cortical ensembles of neurons, the amount of information in the population is limited, yielding nonzero error levels even in the limit of infinitely large populations of neurons. In this letter, we study correlated neuronal populations whose higher-order statistics, and in particular response variances, are also modulated by the stimulus. Weask two questions: Does the correlated noise limit the accuracy of the neuronal representation of the stimulus? and, How can a biological mechanism extract most of the information embedded in the higher-order statistics of the neuronal responses? Specifically, we address these questions in the context of a population of neurons coding an angular variable. We show that the information embedded in the variances grows linearly with the population size despite the presence of strong correlated noise. This information cannot be extracted by linear readout schemes, including the linear population vector. Instead, we propose a bilinear readout scheme that involves spatial decorrelation, quadratic nonlinearity, and population vector summation. We show that this nonlinear population vector scheme yields accurate estimates of stimulus parameters, with an efficiency that grows linearly with the population size. This code can be implemented using biologically plausible neurons. PMID- 15130245 TI - Enhancement of information transmission efficiency by synaptic failures. AB - Many synapses have a high percentage of synaptic transmission failures. I consider the hypothesis that synaptic failures can increase the efficiency of information transmission across the synapse. I use the information transmitted per vesicle release about the presynaptic spike train as a measure of synaptic transmission efficiency and show that this measure can increase with the synaptic failure probability. I analytically calculate the Shannon mutual information transmitted across two model synapses with probabilistic transmission: one with a constant probability of vesicle release and one with vesicle release probabilities governed by the dynamics of synaptic depression. For inputs generated by a non-Poisson process with positive autocorrelations, both synapses can transmit more information per vesicle release than a synapse with perfect transmission, although the information increases are greater for the depressing synapse than for a constant-probability synapse with the same average transmission probability. The enhanced performance of the depressing synapse over the constant-release-probability synapse primarily reflects a decrease in noise entropy rather than an increase in the total transmission entropy. This indicates a limitation of analysis methods, such as decorrelation, that consider only the total response entropy. My results suggest that synaptic transmission failures governed by appropriately tuned synaptic dynamics can increase the information carrying efficiency of a synapse. PMID- 15130246 TI - An algorithm for the detection of faces on the basis of Gabor features and information maximization. AB - We propose an algorithm for the detection of facial regions within input images. The characteristics of this algorithm are (1) a vast number of Gabor-type features (196,800) in various orientations, and with various frequencies and central positions, which are used as feature candidates in representing the patterns of an image, and (2) an information maximization principle, which is used to select several hundred features that are suitable for the detection of faces from among these candidates. Using only the selected features in face detection leads to reduced computational cost and is also expected to reduce generalization error. We applied the system, after training, to 42 input images with complex backgrounds (Test Set A from the Carnegie Mellon University face data set). The result was a high detection rate of 87.0%, with only six false detections. We compared the result with other published face detection algorithms. PMID- 15130247 TI - Analysis of sparse representation and blind source separation. AB - In this letter, we analyze a two-stage cluster-then-l(1)-optimization approach for sparse representation of a data matrix, which is also a promising approach for blind source separation (BSS) in which fewer sensors than sources are present. First, sparse representation (factorization) of a data matrix is discussed. For a given overcomplete basis matrix, the corresponding sparse solution (coefficient matrix) with minimum l(1) norm is unique with probability one, which can be obtained using a standard linear programming algorithm. The equivalence of the l(1)-norm solution and the l(0)-norm solution is also analyzed according to a probabilistic framework. If the obtained l(1)-norm solution is sufficiently sparse, then it is equal to the l(0)-norm solution with a high probability. Furthermore, the l(1)- norm solution is robust to noise, but the l(0)-norm solution is not, showing that the l(1)-norm is a good sparsity measure. These results can be used as a recoverability analysis of BSS, as discussed. The basis matrix in this article is estimated using a clustering algorithm followed by normalization, in which the matrix columns are the cluster centers of normalized data column vectors. Zibulevsky, Pearlmutter, Boll, and Kisilev (2000) used this kind of two-stage approach in underdetermined BSS. Our recoverability analysis shows that this approach can deal with the situation in which the sources are overlapped to some degree in the analyzed domain and with the case in which the source number is unknown. It is also robust to additive noise and estimation error in the mixing matrix. Finally, four simulation examples and an EEG data analysis example are presented to illustrate the algorithm's utility and demonstrate its performance. PMID- 15130248 TI - Minimax mutual information approach for independent component analysis. AB - Minimum output mutual information is regarded as a natural criterion for independent component analysis (ICA) and is used as the performance measure in many ICA algorithms. Two common approaches in information-theoretic ICA algorithms are minimum mutual information and maximum output entropy approaches. In the former approach, we substitute some form of probability density function (pdf) estimate into the mutual information expression, and in the latter we incorporate the source pdf assumption in the algorithm through the use of nonlinearities matched to the corresponding cumulative density functions (cdf). Alternative solutions to ICA use higher-order cumulant-based optimization criteria, which are related to either one of these approaches through truncated series approximations for densities. In this article, we propose a new ICA algorithm motivated by the maximum entropy principle (for estimating signal distributions). The optimality criterion is the minimum output mutual information, where the estimated pdfs are from the exponential family and are approximate solutions to a constrained entropy maximization problem. This approach yields an upper bound for the actual mutual information of the output signals - hence, the name minimax mutual information ICA algorithm. In addition, we demonstrate that for a specific selection of the constraint functions in the maximum entropy density estimation procedure, the algorithm relates strongly to ICA methods using higher-order cumulants. PMID- 15130249 TI - Improving generalization capabilities of dynamic neural networks. AB - This work addresses the problem of improving the generalization capabilities of continuous recurrent neural networks. The learning task is transformed into an optimal control framework in which the weights and the initial network state are treated as unknown controls. A new learning algorithm based on a variational formulation of Pontrayagin's maximum principle is proposed. Under reasonable assumptions, its convergence is discussed. Numerical examples are given that demonstrate an essential improvement of generalization capabilities after the learning process of a dynamic network. PMID- 15130250 TI - A modified algorithm for generalized discriminant analysis. AB - Generalized discriminant analysis (GDA) is an extension of the classical linear discriminant analysis (LDA) from linear domain to a nonlinear domain via the kernel trick. However, in the previous algorithm of GDA, the solutions may suffer from the degenerate eigenvalue problem (i.e., several eigenvectors with the same eigenvalue), which makes them not optimal in terms of the discriminant ability. In this letter, we propose a modified algorithm for GDA (MGDA) to solve this problem. The MGDA method aims to remove the degeneracy of GDA and find the optimal discriminant solutions, which maximize the between-class scatter in the subspace spanned by the degenerate eigenvectors of GDA. Theoretical analysis and experimental results on the ORL face database show that the MGDA method achieves better performance than the GDA method. PMID- 15130251 TI - Stability-based validation of clustering solutions. AB - Data clustering describes a set of frequently employed techniques in exploratory data analysis to extract "natural" group structure in data. Such groupings need to be validated to separate the signal in the data from spurious structure. In this context, finding an appropriate number of clusters is a particularly important model selection question. We introduce a measure of cluster stability to assess the validity of a cluster model. This stability measure quantifies the reproducibility of clustering solutions on a second sample, and it can be interpreted as a classification risk with regard to class labels produced by a clustering algorithm. The preferred number of clusters is determined by minimizing this classification risk as a function of the number of clusters. Convincing results are achieved on simulated as well as gene expression data sets. Comparisons to other methods demonstrate the competitive performance of our method and its suitability as a general validation tool for clustering solutions in real-world problems. PMID- 15130253 TI - Report from the CDC. Vaccines in women. AB - Women's healthcare providers are encouraged to incorporate immunizations into their clients' care. Because women often rely on their healthcare provider for primary and preventive care, that provider may dramatically improve clients' quality of life by decreasing the risk of vaccine-preventable diseases. Women often assume responsibility for the entire family's health, and educating women can prevent disease in the household. Women's healthcare providers should offer and promote these vaccines: hepatitis B, varicella, measles/mumps/rubella, and combined tetanus/diphtheria toxoids for adolescent and young adult women, inactivated influenza vaccine during pregnancy, and pneumococcal, influenza, and tetanus/diphtheria vaccines for the adult or elderly woman. Education should include the importance of vaccines and the rationale for their necessity during each stage of life. Several strategies for implementing and supporting an immunization program have been shown to improve adult immunization rates. These include employing such protocols as standing orders, screening for adult immunizations at each office encounter, and using previously developed immunization documentation forms. The Advisory Committee on Immunization Practices (ACIP) recommendations, vaccine information statements (VIS), and storage and handling guidelines are readily available at low or no cost through CDC and professional organizations or immunization interest group websites. The current adult vaccine schedule assists providers to determine the need for vaccines by displaying graphically both age and medical risk factors. PMID- 15130254 TI - Toward optimal health: the experts discuss abnormal uterine bleeding. PMID- 15130255 TI - Using imiquimod for genital warts in female patients. AB - Genital warts (GW) are the manifestation of infection with specific types of human papillomavirus (HPV), one of the most common sexually transmitted viral infections in the world. Genital warts can be either raised (exophytic) or flat. Exophytic genital warts are most commonly secondary to inoculation of the basal epithelium with nononcogenic HPV types 6 and 11, whereas flat warts are usually secondary to potentially oncogenic HPV types 16 and 31. Genital warts can occasionally cause irritation and discomfort, particularly during intercourse, although most cases are asymptomatic. It is the psychosocial effects of infection, such as a sense of shame, depression, and anxiety, that represent the more significant toll for most patients with GW. Current therapies for GW are mainly ablative and do not directly enhance the immune response to HPV. Therefore, recurrence is a problem for many patients. Imiquimod 5% cream is a patient-applied therapy that directly enhances the immune response to HPV and is safe and effective for the treatment of GW. A reduction in viral load is observed following treatment. Low recurrence rates compared to other treatment modalities may be due to the stimulation of the cell-mediated immune response by imiquimod. PMID- 15130256 TI - Gender-based differences in the prognostic value of coronary calcification. AB - PURPOSE: To investigate the use of electron beam tomography (EBT) screening to predict all-cause mortality in a large cohort of asymptomatic women and men. METHODS AND RESULTS: We obtained mortality information from the National Death Index in 10377 asymptomatic individuals (40% women) referred by primary care physicians for coronary calcification screening. The average follow-up period was 5 +/- 3.5 years. Univariable and multivariable Cox proportional hazard models were developed to predict all-cause mortality. Women had a lower prevalence of coronary calcification and smaller calcification scores than men (p < 0.0001). Death rates were higher among older, diabetic, hypertensive, and currently smoking individuals both in women and in men. In unadjusted (chi-square = 82, p < 0.0001) as well as risk-adjusted (chi-square = 7, p = 0.007) Cox survival models, women had a greater probability of death than men in each strata of calcification. Relative risk (RR) ratios were increased 3.0-fold, 5.5-fold, and 5.5-fold, respectively, for women compared with men with coronary calcification scores of 101-399, 400-1000, and >1000 (p < 0.0001). Using receiver operating characteristics (ROC) curve analyses to assess coronary calcification added incremental prognostic value to Framingham risk scores (p < 0.0001). CONCLUSIONS: In this cohort of asymptomatic women, coronary calcification screening provided incremental prognostic information after adjustment for traditional risk factors. EBT may be a useful tool for risk stratification in women, where the early diagnosis of coronary heart disease (CHD) remains a strong challenge. PMID- 15130257 TI - Adherence to oral therapies in pelvic inflammatory disease. AB - PURPOSE: To assess adherence to oral therapies in pelvic inflammatory disease (PID). METHODS: Medication adherence to oral doxycycline therapy (100 mg) was measured using electronic event monitoring (EEM) among adult women with PID. Subjects (n = 91), who were predominantly black and with a high school or lower educational attainment, were randomly selected from the Pelvic Inflammatory Disease Evaluation and Clinical Health (PEACH) Study, a trial in which subjects were randomized to either (1). initial inpatient then outpatient or (2). fully outpatient treatment with doxycycline and cefoxitin. Summary statistics calculated within treatment groups and in the cohort as a whole included the percentage of administrations taken, percentage of days with correct dosage, percentage of days with no drug taken, time elapsed until the first drug holiday, and percentage of optimal dosing intervals. RESULTS: Study subjects took an average of 70% of prescribed doses, took the prescribed two daily doses for less than half of their outpatient days, took an unscheduled drug holiday for almost 25% of their outpatient days, and took only 16.9% of their doses within the optimal timing interval. In general, working, not bleeding with sex, and not drinking hard liquor were positively associated with measures of adherence. Adherence estimates were similar among women in the inpatient and outpatient groups after hospital discharge. CONCLUSIONS: The disturbing rates of time interval adherence, even after hospitalization, suggest the need to determine the effectiveness of antibiotic regimens involving shorter courses and longer dosing intervals. PMID- 15130258 TI - Efficacy of resistance exercise on lumbar spine and femoral neck bone mineral density in premenopausal women: a meta-analysis of individual patient data. AB - BACKGROUND: Osteoporosis and osteopenia are major public health problems. The purpose of this study was to conduct an individual patient data (IPD) meta analysis to examine the efficacy of resistance exercise on lumbar spine and femoral neck bone mineral density (BMD) in premenopausal women. METHODS: Studies were retrieved via (1). computerized literature searches, (2). review of reference lists from previous studies, (3). hand searching selected journals, and (4). expert review of the reference list. Two x two analysis of covariance (ANCOVA) tests with repeated measures on one factor (time) and study as a covariate were used to analyze changes in BMD. RESULTS: One hundred forty-three subjects (74 exercise, 69 control) were included in the analysis. Changes in lumbar spine BMD averaged 0.006 +/- 0.035 g/cm(2) (0.64 +/- 2.99%) in the exercise group and 0.008 +/- 0.091 g/cm(2) (0.74 +/- 7.58%) in the control group, and changes in femoral neck BMD averaged 0.005 +/- 0.031 g/cm(2) (0.46 +/- 3.10%) in the exercise group and 0.003 +/- 0.031 g/cm(2) (0.31 +/- 2.97%) in the control group. No statistically significant differences in lumbar spine or femoral neck BMD were found within or between the exercise and control groups (p > 0.05). CONCLUSIONS: Based on existing evidence, our results do not support the efficacy of resistance exercise for increasing or maintaining lumbar spine and femoral neck BMD in premenopausal women. PMID- 15130259 TI - The association between physical activity and forearm bone mineral density in healthy premenopausal women. AB - PURPOSE: To analyze the association between recreational and occupational physical activity and forearm bone mineral density (BMD) in healthy premenopausal women. METHODS: During 1984-1986, a population-based health survey (HUNT 1) was conducted among women and men aged >19 years in Nord-Trondelag county in Norway. The second, follow-up survey (HUNT 2) was conducted during 1995-1997. The subjects in this study consist of healthy premenopausal women (n = 1396) < 45 years old in the year of participation of HUNT 2 who had undergone distal and ultradistal radius densitometry in 1995-1997, performed with single-energy x-ray absorptiometry. RESULTS: Women with the highest scores of estimated combined recreational and occupational physical activity (PA) in 1984 and 1995 had significantly higher BMD in the distal radius (mean BMD 0.487 compared with mean BMD 0.480 among those with a low combined PA score) (p for trend = 0.04). At the ultradistal site of the radius, women with a high combined PA score had mean a BMD = 0.403 compared with women with low PA scores (mean BMD = 0.384) (p for trend = 0.017). After adjusting for age, marital status, smoking, amenorrhea, body mass index (BMI), and daily milk consumption, the associations remained the same or got even stronger. CONCLUSIONS: The small group of women in the highest category of PA had a significantly higher forearm BMD and the smallest risk of low BMD. Important unanswered questions remain about the optimal relationship between intensity, amount and type of PA, and BMD and later risk of osteoporosis. Further research on BMD as a surrogate measure of structural and architectural bone quality and the sensitivity of different measuring sites for estimation of the effect of PA on bone is warranted. PMID- 15130260 TI - Posttraumatic stress disorder after pregnancy, labor, and delivery. AB - OBJECTIVES: Other studies of posttraumatic stress disorder (PTSD) after birth did not include questions about prior traumatic life events. This study sought to determine if a difficult birth was associated with symptoms of PTSD as well as considering sociodemographics, history of violence, depression, social support, and traumatic life events. METHODS: New mothers were recruited on the postpartum ward of six Toronto-area hospitals (n = 253) and were interviewed by telephone 8 10 weeks postpartum (n = 200). We dichotomized the postpartum stress (PTS) into high PTS (answered "yes" to 3 or more items) or low PTS (answered "yes" to 0-2 items). We calculated the odds ratios between difficult birth, other factors, and the binary PTS variable. RESULTS: Results of multivariable logistic regression revealed that no factor suggestive of a difficult birth was significantly related to high PTS scores, except having two or more maternal complications (odds ratio [OR] = 4.0, 95% confidence interval [CI] = 1.3-12.8). Other independent predictors of high PTS scores were depression during pregnancy (OR = 18.9, 95% CI = 5.8-62.4), having two or more traumatic life events (OR = 3.2, 95% CI = 1.2 8.3), being Canadian born (OR = 3.2, 95% CI = 1.3-8.1), and having higher household income (lowest income group, OR = 0.1, 95% CI = 0.02-0.5), intermediate income group OR = 0.4, 95% CI = 0.2-0.8). CONCLUSIONS: In this study, postpartum stress symptoms appeared to be related more to stressful life events and depression than to pregnancy, labor, and delivery. PMID- 15130261 TI - Hormone replacement, race, and psychological health in women: a report from the NHLBI-Sponsored WISE Study. AB - PURPOSE: We analyzed the impact of hormone replacement therapy (HRT) on psychological factors in white and black women. We hypothesized that both groups of women would have fewer symptoms of depression and lower hostility scores associated with HRT use. METHODS: The cohort included 463 postmenopausal women from the National Heart, Lung and Blood Institute (NHLBI)-sponsored Women's Ischemia Syndrome Evaluation (WISE) study. WISE is a four-center study of women with chest pain who underwent quantitative coronary angiography for suspected ischemia. The psychosocial indices included the Beck Depression Inventory (BDI) and the Cook Medley Hostility questionnaire measuring cynicism, hostility, and aggression. RESULTS: There were no differences by race in use, duration, and type of HRT or presence of menopausal symptoms. There were differences by race in baseline psychological measurements, with black women exhibiting higher BDI scores and higher total Cook Medley scores (p = 0.03) than white women. Use of HRT was consistently associated with better psychological health in white women, with fewer symptoms of depression and lower aggression and cynicism scores (p < 0.04). Black women with menopausal symptoms who used HRT had significantly lower hostility (p < 0.01) and cynicism scores (p < 0.05) than black women who did not use HRT. The presence of menopausal symptoms and hysterectomy status were significant independent predictors of HRT use for both white and black women (p < 0.05). CONCLUSIONS: We observed racial differences in associations between HRT use and psychological health. Within the white but not the black HRT users, there were fewer symptoms of depression and lower aggression and cynicism scores. PMID- 15130262 TI - Midlife women's responses to a hospital sleep challenge: aging and menopause effects on sleep architecture. AB - OBJECTIVE: To distinguish aging from menopause effects on sleep architecture, we studied an episode of disturbed hospital sleep in asymptomatic midlife women during the follicular phase of an ovulatory cycle and three control groups differing by age or menopause status. METHODS: Fifty-one studies were conducted in four groups of volunteers: young cycling (YC, 20-30 years, n = 14), older cycling (OC, 40-50 years, n = 15), ovariectomized receiving estrogen therapy (OVX, 40-50 years, n = 12), and spontaneously postmenopausal (PM, 40-50 years, n = 10). Subjects were admitted to the University Hospital General Clinical Research Center (GCRC) for a first-night sleep study conducted during a 24-hour, frequent blood sampling protocol. RESULTS: Despite similar estrogen concentrations in the YC (28 +/- 4 pg/ml) and OC (34 +/- 6 pg/ml) groups, OC women had reduced sleep efficiency (79% +/- 2%) vs. YC (87% +/- 3%; p = 0.009). In the OVX and PM groups where estrogen concentrations were markedly different, sleep efficiency was also reduced vs. the YC group (OVX vs. YC, 79% +/- 3% vs. 87% +/- 3%, p = 0.05; PM vs. YC, 75% +/- 3% vs. 87% +/- 3%, p = 0.007). Wake time was longer in the three older groups (103 +/- 10 minutes, 101 +/- 12 minutes, 123 +/- 12 minutes for OC, OVX, PM, respectively) vs. YC (63 +/- 13 minutes, p < 0.05). The number of stage shifts was positively associated with advancing age (rho = 0.3, p < 0.03) but not with estrogen concentration. CONCLUSIONS: Aging related sleep deficits in response to an experimental stressor occur in midlife women prior to menopause. PMID- 15130263 TI - Gender differences in time spent during direct observation of doctor-patient encounters. AB - BACKGROUND: Despite increasing recognition of women's health needs, little is known about how primary care physicians spend time with women. Therefore, we examined differences in time use and preventive service delivery during outpatient visits by male and female patients. METHODS: As part of a multimethod study of 138 family physicians, 3384 outpatient visits by adults were directly observed, medical records were reviewed, and patient surveys were performed. Time use was assessed by the Davis Observation Code, which classifies every 15 seconds into 20 behavioral categories. Receipt of health habit counseling recommended by the U.S. Preventive Services Task Force was assessed by direct observation, and eligibility was determined by chart review. Logistic regression and multivariate analysis of variance (ANOVA) were used to compare time use and preventive service delivery in visits by women vs. men. RESULTS: Sixty-four percent of adult visits were from women. Women reported poorer physical health, had higher rates of anxiety (12.5% vs. 7.4% in men), and depression (21.9% vs. 8.4% in men), a higher percent of visits for well care (10.2% vs. 8.8% in men), and more drugs prescribed (64.8% vs. 61% in men) and raised more emotional issues than men (14.7% vs. 7.5%). After controlling for visit and patients characteristics, visits by women had a higher percent of time spent on physical examination, structuring the intervention, patient questions, screening, and emotional counseling. Visits by men involved a higher percent of time spent on procedures and health behavior counseling. More eligible men than women received exercise, diet, and substance abuse counseling. Patients of female physicians exhibited gender differences in only one category of how time was spent (substance abuse), whereas among patients of male physicians, gender differences were noted in 10 of the 20 categories. CONCLUSIONS: Outpatient visits by women differ from those of men in ways that reflect women's unique healthcare needs but also raise concern about unequal delivery of health habit counseling for diet and exercise. PMID- 15130266 TI - Hepatocellular carcinoma: paradigm of preventive oncology. AB - Morbidity and mortality from hepatocellular carcinoma (HCC), which is primarily caused by hepatitis B virus or hepatitis C virus, can be prevented. Public health interventions have eliminated transfusion transmission of these viruses and, in endemic countries with effective hepatitis B virus vaccination programs, have greatly reduced incident hepatitis B virus infections (and HCC) in children. Antiviral treatment can eliminate detectable hepatitis C virus in 50%-80% of chronically infected patients, presumably reducing their risk of cancer. HCC survival rates remain universally poor, but early detection and treatment in developed countries has improved survival in selected patients. Despite these advances, worldwide HCC rates remain high, and additional preventive efforts are needed. The most important opportunity is wider distribution of hepatitis B virus vaccine in endemic areas. Development of an HCV vaccine, improved antiviral therapies, and better methods for HCC detection would also help decrease morbidity and mortality from HCC. HCC prevention efforts provide a paradigm for preventive oncology in cancers of viral etiology. PMID- 15130267 TI - Advances in the surgical management of liver malignancies. AB - Primary malignancies of the liver include tumors arising from the hepatocytes (hepatocellular carcinoma and the fibrolamellar variant) and the intrahepatic bile ducts (intrahepatic cholangiocarcinoma). Hepatocellular carcinoma is the most common primary cancer of the liver and is a leading cause of death from cancer worldwide. Although it is uncommon in the United States, the incidence of hepatocellular carcinoma is rising. Hepatitis, ethanol use, and cirrhosis often dominate the clinical picture and may dictate prognosis. New clinical and pathological staging systems have allowed for the more accurate stratification of patients to more appropriately identify patients for resection, transplantation, and percutaneous ablation therapies. A correlation between liver volume and surgical outcome has recently been demonstrated, with small liver remnant size being associated with increased morbidity. Portal vein embolization has therefore been proposed as one way to induce hypertrophy of the anticipated liver remnant before resection. Initial reports have shown that portal vein embolization decreases the incidence of postoperative complications. More recently, systemic chemotherapy and chemoembolization have been investigated as both primary and neoadjuvant therapy. Chemoimmunotherapy with 5-fluorouracil and interferon may be associated with a superior response rate in the fibrolamellar variant of hepatocellular carcinoma. Two recent randomized studies have also indicated improved survival after hepatic artery embolization in selected patients. PMID- 15130268 TI - Emerging role of transplantation for primary liver cancers. AB - Liver transplantation (LT) has been utilized in the treatment of primary hepatic malignancy for decades. Hepatocellular cancer (HCC) remains the most common malignant condition treated with LT, with almost 400 such transplants performed annually in the US. Refinement in the selection criteria for LT in patients with HCC has led to survival rates similar to those for LT in nonmalignant conditions. Excellent results have also been reported following LT for select patients with epithelioid hemangioendothelioma and hepatoblastoma. Patients with cholangiocarcinoma treated with LT have generally faired poorly, with survival rates far below that of LT for nonmalignant conditions. Improved survival has recently been reported following LT for cholangiocarcinoma in highly select patients treated with aggressive neoadjuvant therapy. The future utility of LT in the treatment of malignancy will be influenced by several factors, including a profound organ donor shortage faced worldwide; increasing prevalence of hepatitis C, HCC and cirrhosis; and the evolution of live donor liver transplantation. PMID- 15130269 TI - Novel systemic therapy options for hepatocellular carcinoma. AB - Hepatocellular carcinoma, a common malignancy globally, has been increasing in incidence in the United States, mostly due to the rising incidence of Hepatitis C viral (HCV) infection. The prognosis of patients with this cancer has been poor and even tumor resection has rarely been curative. However, orthotopic liver transplantation (OLT) has been associated with long-term survival benefit and cures, provided rigorous patient-selection criteria were adhered to. Liver cirrhosis is the most common precursor for HCC, and attempts have been made to prevent the progression from liver cirrhosis to HCC. Post resection adjuvant therapies have included interferon, polyprenoic acid, and adoptive immunotherapy. Finding effective systemic treatments for non-resectable HCC has been challenging and quite frustrating. The presence of liver cirrhosis and the associated volume expansion, electrolyte imbalances, decreased liver synthetic and metabolic reserve, and portal hypertension has made the design of systemic therapy for HCC a major challenge. Additionally staging of HCC using the Tumor Node Metastases (TNM) system, but ignoring the underlying liver disease made it extremely difficult to compare results of different trials. However by and large it would seem, that the more aggressive chemotherapy agents and combinations were associated with median survival times of 3-5 months. Considering the vascular nature of HCC it may be reasonable to combine tolerable chemotherapy with newly released agents with angiogenesis inhibiting properties. Thus, systemic therapy of HCC is a work in progress that calls for additional trials of tolerable newer agents and combinations. PMID- 15130270 TI - Recent advances in CT imaging of liver metastases. AB - Multidetector computed tomography provides robust evaluation of the hepatic parenchyma. It plays a critical role in the detection of liver metastases and the assessment of treatment response to therapy. In this article, we discuss the role of multidetector computed tomography in the detection and characterization of hepatic metastases, and the value of image processing with volume rendering and maximum-intensity projection techniques in the accurate delineation of hepatic vascular anatomy and the segmental localization of lesions. This information is critical in the diagnosis and treatment of patients with metastatic disease and is essential in surgical and nonsurgical planning. PMID- 15130271 TI - Recent advances in imaging hepatocellular carcinoma: diagnosis, staging and response assessment: functional imaging. AB - Historically, nuclear medicine has had an important role in the differential diagnosis of liver tumors but has been largely superseded by other forms of conventional imaging, in particular computed tomographic portography. It remains helpful in difficult cases because it has characteristic features in both hepatocellular carcinoma (HCC) and benign conditions. 131I is an important therapeutic tool. FDG-PET is useful in certain cases, especially for finding metastases and monitoring response to therapy. PMID- 15130272 TI - Current role of hepatic artery infusion and isolated liver perfusion for the treatment of colorectal cancer liver metastases. AB - There are many treatment options for patients with metastatic colorectal carcinoma confined to the liver. Surgical resection alone can result in significant prolongation of survival in patients with favorable prognostic factors. Randomized studies of hepatic artery infusion therapy after complete resection of liver metastases have demonstrated improvements in hepatic recurrence-free survival but no impact on overall survival. Randomized trials evaluating the treatment of unresectable disease with hepatic artery infusion therapy have demonstrated higher response rates (31%-50%) than those seen with systemic chemotherapy (8%-20%) but no survival benefit. Vascular isolation and perfusion of the liver with chemotherapy with or without biologic agents under hyperthermic conditions is another regional modality that has been explored for the treatment of unresectable colorectal cancer liver metastases. Large series report high partial response rates (68%-77%), with responses being achieved in patients with advanced tumor burden and in those who have disease progression through prior treatment of hepatic metastases. PMID- 15130273 TI - The role of radiotherapy in the treatment of liver metastases. AB - Radiotherapy has historically played a minor role in the treatment of patients with unresectable liver metastases from colorectal cancer and other malignancies. This can be attributed chiefly to the low tolerance of the whole liver to radiation. High-precision radiotherapy planning techniques have allowed much higher doses of radiation to be delivered safely to focal liver metastases, while sparing most of the normal liver. When combined with hepatic arterial fluorodeoxyuridine, high-dose focal liver radiotherapy is associated with excellent response rates, local control, and survival in patients with unresectable liver metastases from colorectal cancer. Radiotherapy, with and without concurrent systemic chemotherapy, has also been used with encouraging outcomes for patients with liver metastases from colorectal cancer and other cancers. There appears to be a radiation dose response for liver metastases; tumors treated with doses of 70 Gy or greater are likelier to have durable local control. Advancements in tumor imaging, in radiotherapy techniques that will allow the safe delivery of higher doses of radiation, and in novel tumor radiation sensitizers and normal tissue radioprotectors should substantially improve the outcome of patients with unresectable liver metastases treated with radiotherapy. PMID- 15130274 TI - Redox control of carcinogenesis and tumor biology. PMID- 15130275 TI - Mitochondria, manganese superoxide dismutase, and cancer. PMID- 15130276 TI - Reversible modulation of cell cycle kinetics in NIH/3T3 mouse fibroblasts by inducible overexpression of mitochondrial manganese superoxide dismutase. AB - To study the mechanism(s) by which manganese-containing superoxide dismutase (MnSOD) mediates cellular growth inhibition, an inducible retroviral vector system regulated by the lac repressor was used to overexpress MnSOD protein in NIH/3T3 cells. Increased MnSOD activity led to decreased cell growth due to prolonged cell cycle transition times in G(1) and S phases without significant changes in G(2)/M phase. Changes in cell cycle transition time were reversible and tightly correlated with MnSOD levels. A transient increase of reactive oxygen species and concomitant decrease in mitochondrial membrane potential were documented following MnSOD induction. N-Acetyl-L-cysteine prevented growth inhibition by MnSOD. Our data suggest that MnSOD may serve a physiological function of regulating cell cycle progression through its prooxidant activity of generating hydrogen peroxide, resulting in coordination of mitochondrial redox state and cellular proliferation. PMID- 15130277 TI - Tumor suppressive effects of MnSOD overexpression may involve imbalance in peroxide generation versus peroxide removal. AB - Manganese superoxide dismutase (MnSOD) activity is generally lower in cancer cells when compared with their normal cell counterparts. Many studies have shown that replacing the diminished MnSOD activity leads to inhibition of the malignant phenotype. We sought to overexpress MnSOD in a chemically transformed, malignant rat cell line with low endogenous MnSOD activity to determine the effect on the malignant phenotype. After MnSOD cDNA transfection, clonal populations were characterized at the molecular level for protein, RNA, and DNA, as well as for in vitro and in vivo growth and in vivo lung metastasis. MnSOD transfectants, which both under- and overexpressed MnSOD protein, were identified. These transfectants demonstrated variations in glutathione peroxidase and catalase activity levels, indicating differences in peroxide-generating versus peroxide-metabolizing enzymes (antioxidant imbalance); these differences were suggestive of alterations in their abilities to metabolize peroxide when compared with the parental cell line. In addition, these transfectants demonstrated reductions in both in vitro and in vivo growth, as well as a reduction in metastatic potential, which correlated with antioxidant imbalance. These results suggest that the tumor suppressive effect of MnSOD overexpression is in part mediated by an antioxidant imbalance resulting in the reduced capacity to metabolize increased levels of intracellular peroxides. PMID- 15130278 TI - Alteration of cellular phenotype and responses to oxidative stress by manganese superoxide dismutase and a superoxide dismutase mimic in RWPE-2 human prostate adenocarcinoma cells. AB - To study biologic effects of increased manganese superoxide dismutase (MnSOD) on cell behavior, we overexpressed MnSOD in a human prostate cancer cell line RWPE-2 by cDNA transfection. Stable transfectants of MnSOD showed a two- to threefold increase in MnSOD protein and enzymatic activity and a decrease in growth rate with prolonged cell population doubling times. Western blot analysis showed a 1.5 to twofold increase in the cyclin-dependent kinase inhibitor p21(Waf1) in MnSOD transfectants. Overexpression of MnSOD resulted in a seven- to eightfold increase in reduced glutathione (GSH), 18- to 26-fold increase in oxidized glutathione (GSSG), and a two- to threefold decrease in the ratio of GSH to GSSG. MnSOD overexpressing cells showed an increase in sensitivity to the cytotoxicity of buthionine sulfoximine, a glutathione-depleting agent, and vitamin C, but a decrease in sensitivity to sodium selenite. Treatment with a superoxide dismutase (SOD) mimic MnTMPyP resulted in similar effects of MnSOD overexpression on cell responses to vitamin C and selenium. These data demonstrate that overexpression of MnSOD or treatment with SOD mimics can result in antioxidant or prooxidant effects in cells, depending on the presence of other antioxidants and prooxidants. MnSOD also has redox regulatory effects on cell growth and gene expression. These findings suggest that MnSOD and SOD mimics have the potential for cancer prevention or treatment. PMID- 15130279 TI - Mitochondrial signal lacking manganese superoxide dismutase failed to prevent cell death by reoxygenation following hypoxia in a human pancreatic cancer cell line, KP4. AB - One of the major characteristics of tumor is the presence of a hypoxic cell population, which is caused by abnormal distribution of blood vessels. Manganese superoxide dismutase (MnSOD) is a nuclear-encoded mitochondrial enzyme, which scavenges superoxide generated from the electron-transport chain in mitochondria. We examined whether MnSOD protects against hypoxia/reoxygenation (H/R)-induced oxidative stress using a human pancreas carcinoma-originated cell line, KP4. We also examined whether MnSOD is necessarily present in mitochondria to have a function. Normal human MnSOD and MnSOD without a mitochondrial targeting signal were transfected to KP4 cells, and reactive oxygen species, nitric oxide, lipid peroxidation, and apoptosis were examined as a function of time in air following 1 day of hypoxia as a H/R model. Our results showed H/R caused no increase in nitric oxide, but resulted in increases in reactive oxygen species, 4-hydroxy-2 nonenal protein adducts, and apoptosis. Authentic MnSOD protected against these processes and cell death, but MnSOD lacking a mitochondrial targeting signal could not. These results suggest that only when MnSOD is located in mitochondria is it efficient in protecting against cellular injuries by H/R, and they also indicate that mitochondria are primary sites of H/R-induced cellular oxidative injuries. PMID- 15130280 TI - In situ reduction of oxidative damage, increased cell turnover, and delay of mitochondrial injury by overexpression of manganese superoxide dismutase in a multistage skin carcinogenesis model. AB - To study early subcellular pathologic changes of tumorigenesis in mouse skin and possible modulation by overexpression of the mitochondrial antioxidant enzyme manganese superoxide dismutase (MnSOD), skin keratinocytes from nontransgenic (Ntg) and transgenic (TgH) mice overexpressing MnSOD topically treated with one dose of 7,12-dimethylbenz(a)anthracene (DMBA) and a subsequent dose of 12-O tetradecanoylphorbol 13-acetate (TPA) were analyzed in situ for levels of MnSOD and the oxidative damage product 4-hydroxy-2-nonenal (4HNE)-modified proteins using specific antibodies and immunogold electron microscopy. At all selected time points analyzed after TPA treatment, there was more MnSOD immunoreactive protein in mitochondria of keratinocytes of TgH mice than Ntg mice. Compared with untreated groups, there was a large increase in 4HNE-modified proteins at 6-24 h after TPA treatment, and this increase was larger in Ntg than TgH mice. Indices of mitosis and apoptosis of keratinocytes were greater in DMBA/TPA-treated TgH than Ntg mouse skin. Mitochondrial injury detected by transmission electron microscopy was delayed in keratinocytes of TgH compared with Ntg mice. The present study demonstrated that overexpression of MnSOD not only protected cells from oxidative damage, but also affected cell turnover kinetics. Thus, previously identified reduction in papilloma formation observed in TgH mice is correlated with mitochondrial events. PMID- 15130281 TI - Amelioration of oxidative stress with ensuing inflammation contributes to chemoprevention of H. pylori-associated gastric carcinogenesis. AB - The gastric inflammatory response provoked by Helicobacter pylori (H. pylori) consists of infiltrations by neutrophils, lymphocytes, and macrophages, resulting in varying degrees of epithelial cell damage. H. pylori-associated inflammation not only activates various oxidant-producing enzymes such as NADPH oxidase and inducible nitric oxide synthase, but also lowers the antioxidant ascorbic acid in the stomach. Reactive oxygen metabolites and nitrogen metabolites generated by these enzymes react with each other to generate new or more potent reactive species. The specific types of cellular damage resulting from reactive oxygen metabolites include lipid peroxidation, protein oxidation, and oxidative DNA damage. All of these oxidative products can result in biochemical changes leading to cancer. A positive association has been demonstrated between H. pylori infection and gastric adenocarcinoma with increased oxidative stress. Therefore, appropriate treatment to reduce oxidative stress would be expected to prevent subsequent gastric carcinogenesis through lessening of H. pylori-associated inflammation. This review will provide evidence that antiinflammatory regimens can decrease the development of tumors and the amelioration of gastric inflammation might lead to chemoprevention strategies by the attenuation of oxidative stress. PMID- 15130282 TI - Role of reactive oxygen species in skin carcinogenesis. AB - Reactive oxygen species (ROS) are associated not only with initiation, but also with promotion and progression in the multistage carcinogenesis model. In the present review, we will focus on the involvement of ROS in skin carcinogenesis, especially that induced by ultraviolet (UV) radiation. UV-specific DNA damage has been well studied thus far. However, recent reports have revealed the previously unknown participation of oxidative stress in UV-induced skin carcinogenesis. Indeed, in addition to transition-type mutations at dipyrimidine sites, G:C to T:A transversions, which may be induced by the presence of 8-oxoguanine during DNA replication, are frequently observed in the ras oncogene and p53 tumor suppressor gene in human skin cancers of sun-exposed areas and in UV-induced mouse skin cancers. Recent studies have shown that not only UV-B, but also UV-A is involved in UV-induced carcinogenesis. A wide variety of biological phenomena other than direct influence by UV, such as inflammatory and immunological responses and oxidative modifications of DNA and proteins, appear to play roles in UV-induced skin carcinogenesis. Furthermore, it has become clear that genetic diseases such as xeroderma pigmentosum show deficient repair of oxidatively modified DNA lesions. The involvement of ROS in skin carcinogeneisis caused by arsenic and chemical carcinogens will also be discussed. PMID- 15130283 TI - Antioxidants of the beverage tea in promotion of human health. AB - Tea that contains many antioxidants is a pleasant and safe drink that is enjoyed by people across the globe. Tea leaves are manufactured as black, green, or oolong. Black tea represents approximately 78% of total consumed tea in the world, whereas green tea accounts for approximately 20% of tea consumed. The concept of "use of tea for promotion of human health and prevention and cure of diseases" has become a subject of intense research in the last decade. Diseases for which tea drinkers appear to have lower risk are simple infections, like bacterial and viral, to chronic debilitating diseases, including cancer, coronary heart disease, stroke, and osteoporosis. Initial work on green tea suggested that it possesses human health-promoting effects. In recent years, the research efforts have been expanded to black tea as well. Research conducted in recent years reveals that both black and green tea have very similar beneficial attributes in lowering the risk of many human diseases, including several types of cancer and heart diseases. For cancer prevention, evidence is so overwhelming that the Chemoprevention Branch of the National Cancer Institute has initiated a plan for developing tea compounds as cancer-chemopreventive agents in human trials. Thus, modern medical research is confirming the ancient wisdom that therapy of many diseases may reside in an inexpensive beverage in a "teapot." PMID- 15130284 TI - EPR spectroscopy in biology and medicine. PMID- 15130285 TI - Factors influencing nitroxide reduction and cytotoxicity in vitro. AB - Nitroxides have been shown to be effective antioxidants, radiation protectors, and redox-active probes for functional electron paramagnetic resonance (EPR) imaging. More recently, the nitroxide 4-hydroxy-2,2,6,6-tetramethylpiperidinyl-N oxyl (Tempol) has been shown to exert differential cytotoxicity to tumor compared with normal cell counterparts. Nitroxides are readily reduced in tissues to their respective hydroxylamines, which exhibit less cytotoxicity in vitro and do not provide radiation protection or an EPR-detectable signal for imaging. In order to better understand factors that influence nitroxide reduction, the rate of reduction of Tempol in mouse and human cell lines and in primary cultures of tumor cells was measured using EPR spectroscopy. Additionally, the cytotoxicity of high concentrations of Tempol and the hydroxylamine of Tempol (Tempol-H) was evaluated in wild-type and glucose-6-phosphate dehydrogenase (G6PD)-deficient Chinese hamster ovary cells. The results show that in general Tempol was reduced at a faster rate when cells were under hypoxic compared with aerobic conditions. Neither depletion of intracellular glutathione nor treatment of cells with sodium cyanide influenced Tempol reduction rates. G6PD-deficient cells were found to reduce Tempol at a significantly slower rate than wild-type cells. Likewise, Tempol-induced cytotoxicity was markedly less for G6PD-deficient cells compared with wild-type cells. Tempol-H exhibited no cytotoxicity to either cell type. Tempol-mediated cytotoxicity was enhanced by glutathione depletion and inhibition of 6-phosphogluconate dehydrogenase in wild-type cells, but was unaltered in G6PD deficient cells. Collectively, the results indicate that while the bioreduction of Tempol can be influenced by a number of factors, the hexose monophosphate shunt appears to be involved in both nitroxide reduction as well as cytotoxicity induced by high levels of exposure to Tempol. PMID- 15130286 TI - Simultaneous measurement of oxygenation in intracellular and extracellular compartments of lung microvascular endothelial cells. AB - A new technique is described for simultaneous determination of intra- and extracellular oxygen concentrations [partial pressure of O(2) (pO(2))] in bovine lung microvascular endothelial cells (BLMVECs) using electron paramagnetic resonance (EPR) oximetry. The measurements were performed in BLMVEC suspensions of a 20- micro l volume containing 4,000 cells. The extracellular pO(2) was measured using a trityl EPR probe [triarylmethyl (TAM), 10 micro M], a tricarboxylate anion radical, that stays exclusively in the extracellular space. The intracellular oxygen was measured using a pre-internalized particulate spin probe, lithium 5,9,14,18,23,27,32,36-octa-n-butoxy-naphthalocyanine (LiNc-BuO). Because there is a wide discrepancy in the reported values of cellular oxygenation by and large due to differences in the methods employed, we utilized the dual EPR probe technique to measure the oxygen gradient that apparently exists across the cell membrane. The intra- and extracellular pO(2) values were 139 +/- 2.5 and 157 +/- 3.6 mm Hg, respectively, for cells exposed to room air. A fairly smaller gradient of oxygen was observed in cells exposed to 7.5% oxygen (pO(2) = 57 mm Hg). In summary, this study confirms the feasibility of simultaneous and accurate measurements of intra- and extracellular pO(2) using LiNc-BuO and TAM EPR oximetry probes. PMID- 15130287 TI - In vivo measurement of redox status in streptozotocin-induced diabetic rat using targeted nitroxyl probes. AB - In vivo electron paramagnetic resonance (EPR) with nitroxyl spin probes has been used for the evaluation of in vivo free radical reactions and redox status in living animals. The aim of this study was to clarify the location of free radical reactions induced by hyperglycemia in osteogenic disorder shionogi (ODS) rats using in vivo EPR spectroscopy. Diabetes was induced by intravenous injection of streptozotocin (STZ). The amount of ascorbic acid (AsA) in ODS rats was controlled by feeding AsA-containing water. Fourteen days after STZ injection, blood glucose and plasma malondialdehyde levels in STZ-treated rats significantly increased compared with untreated rats. Signal decay rates of intravenously injected 3-carbamoyl-2,2,5,5-tetramethyl-1-pyrrolidinyloxy (carbamoyl-PROXYL) (less membrane permeable) and 3-carboxy-PROXYL (membrane impermeable) were enhanced in STZ-treated rats in agreement with the previous reports. The decay rate of 3-acetoxymethoxy-PROXYL (membrane permeable) was significantly enhanced by STZ treatment in AsA-depleted rats, and this enhancement was partially restored to the control value by xanthine oxidase inhibitor, although the rate in AsA-supplemented rats was not changed by STZ treatment. These results suggested that the enhancement of signal decay occurred mainly in the intravascular region in STZ-induced diabetic rats and that AsA depletion induced the enhancement of intracellular signal decay through xanthine oxidase, although it is not clear whether the enhancement of signal decay is the cause or the effect of STZ-induced diabetes. PMID- 15130288 TI - Nitric oxide as a prognostic marker for neurological diseases. AB - The potential value of the nitric oxide (NO) level as a prognostic marker in human brain diseases is investigated. Cerebrospinal fluid (CSF) collected from neurological patients was examined for NO content using electron paramagnetic resonance (EPR) spectroscopy. In adult patients with meningitis, the level of NO was higher than that in other groups of brain disorders, such as brain traumas and brain tumors. Very high levels of NO in the CSF appeared to be correlated with a high incidence of fatal outcomes. In children with meningitis, it was possible to differentiate between viral and bacterial origin of the disease as evidenced by the EPR analysis of the CSF. The results indicated that NO levels in the CSF can be a useful prognostic marker in neurological diseases. PMID- 15130289 TI - Detection of reactive oxygen and nitrogen species by EPR spin trapping. AB - Electron paramagnetic resonance spin trapping has become an indispensable tool for the specific detection of reactive oxygen free radicals in biological systems. In this review we describe some of the advantages as well as some experimental considerations of this technique and how it can be applied to biological systems to measure oxidative stress. PMID- 15130290 TI - Detection of lipid radicals using EPR. AB - Cells oxidize molecules to generate energy and to make the materials to build and support the structures and functions needed for life. However, unwanted oxidations can damage these same structures and impair function. Lipids (the lipids in membranes and lipoproteins) are targets of unwanted oxidations. The primary mechanism of these oxidations is free radical-mediated chain reactions. Here we provide an overview of how electron paramagnetic resonance (EPR) can be used to detect the free radicals formed during lipid peroxidation. Although direct detection of lipid-derived radicals has been accomplished, the approach is not feasible for detecting these radicals in cells. Spin trapping with alpha-(4 pyridyl-1-oxide)-N-tert-butylnitrone and 5,5-dimethyl-pyrroline-1-oxide has provided the most information on cellular lipid peroxidation. We present some considerations for successful detection of lipid radicals by EPR. PMID- 15130291 TI - Spin trapping of nitric oxide with the iron-dithiocarbamate complex: chemistry and biology. AB - This brief review describes chemical and biological aspects concerning spin trapping of nitric oxide (NO) with the iron-dithiocarbamate (Fe-DTC) complex as a spin trap. Knowledge on basic properties of the Fe-DTC complex would help in understanding the applicability and limitation of the Fe-DTC-based NO spin trapping method when it is employed in viable biological systems. PMID- 15130292 TI - In vivo spin trapping of nitric oxide. AB - The measurement of nitric oxide (NO) in biological samples has normally required destructive chemical techniques. The ability to detect NO non-invasively in living animals or excised organs has great potential using specialized electron paramagnetic resonance (EPR) methods. Although NO is paramagnetic, it cannot be observed directly unless it is complexed with ferrous iron-dithiocarbamate ligand spin trap complexes. Despite the minimally invasive nature of the technique, highly sensitive localized concentrations of NO may be observed ("trapped") in vivo by both L-band EPR and magnetic resonance imaging. PMID- 15130293 TI - Redox reactions of hemoglobin. AB - Redox reactions of hemoglobin have gained importance because of the general interest of the role of oxidative stress in diseases and the possible role of red blood cells in oxidative stress. Although electron paramagnetic resonance (EPR) is extremely valuable in studying hemoglobin redox reactions it has not been adequately used. We have focused in this review on the important contributions of EPR to our understanding of hemoglobin redox reactions. We have limited our discussion to the redox reactions thought to occur under physiological conditions. This includes autoxidation as well as the reactions of hydrogen peroxide generated by superoxide dismutation. We have also discussed redox reactions associated with nitric oxide produced in the circulation. We have pinpointed the value of using EPR to detect and study the paramagnetic species and free radicals formed during these reactions. We have shown how EPR not only identifies the paramagnetic species formed but can also be used to provide insights into the mechanism involved in the redox reactions. PMID- 15130294 TI - In vitro and in vivo measurement of pH and thiols by EPR-based techniques. AB - In vitro and in vivo measurements of pH and thiols provide critical information on physiology and pathophysiology of living organisms, particularly related to oxidative stress. Stable nitroxides of imidazoline and imidazolidine types provide the unique possibility of measuring local values of pH and glutathione content in various biological systems, including in vivo studies. The basis for these applications is the observation of specific chemical reactions of these nitroxides with protons or thiols, followed by significant changes in the electron paramagnetic resonance (EPR) spectra of these probes, measured by low frequency EPR techniques. The applications of some newly developed pH and SH probes in model systems of pharmacological interest, biological fluids, tissues, and cells as well as in vivo studies in isolated hearts and in the gut of living animals are discussed. PMID- 15130295 TI - Using EPR to measure a critical but often unmeasured component of oxidative damage: oxygen. AB - Oxygen is a critical variable in oxidative damage. It can be a direct reactant in one or more of the pertinent reactions that result in oxidative damage. It also is an essential substrate for mitochondrial respiration and many other essential synthetic and degradative reactions. The level of oxygen can have a regulatory role, affecting the rate and direction of metabolic processes and physiological functions that are germane to the pathophysiological processes that are being studied. Its supply to tissue and to cells is therefore a critical parameter governing normal homeostasis. The level of oxygen at specific sites may affect cell signaling. It therefore seems clear that it can be very useful to measure oxygen when studying oxidative damage. In order for the measurements of oxygen to be most useful, it often is essential to measure the amount of oxygen at particular sites and under appropriate conditions. These needs require methodology that can make sensitive and localized measurements of oxygen. Electron paramagnetic resonance (EPR) oximetry is such a technique, plus it has the capability of making repeated measurements from the same site non-invasively. The principles and applications of EPR oximetry to viable systems, including cell suspensions and intact animals, are described in this paper. PMID- 15130296 TI - [Excited achievement from intracranial tumor research]. PMID- 15130297 TI - [Brain death and organ donation: the ultimate combination of modern science and humanism]. PMID- 15130299 TI - [Cluster analysis and identify significance of novel genes related to glioma in molecule classification]. AB - OBJECTIVE: To screen differentially expressed genes in the development of human glioma and establish molecular classification of glioma preliminary based on gene expression using cDNA microarray. METHODS: Brain specimens were obtained from 18 patients with glioma, 10 males and 8 females, aged 14 approximately 62 with an average age of 44.4. The total RNAs of these glioma specimens and 2 specimens of donated brain of normal adults were extracted. BioStarH140S microarray (including 8347 old genes and 5592 novel genes) were adopted and hybridized with probes which were prepared from the total RNAs. Differentially expressed genes between the normal tissues and glioma tissues were assayed after scanning cDNA microarray with ScanArray 4000. Northern hybridization, and in situ hybridization (ISH) were used to identify the functions of novel genes. Those differentially expressed genes were studied with Hierarchical method and molecular classification of glioma was preliminarily carried out. RESULTS: Among the 13 939 target genes, there were 1200 (8.61%) differentially expressed genes and 395 (2.83%) novel genes. 348 genes were up-regulated and 852 genes were down-regulated in glioma. The results of bioinformatical analysis, Northern hybridization and ISH revealed that those novel genes were highly associated with glioma. There were multiple genes which were relevance to classification by Hierarchical method, such as MAP gene, cytoskeleton and matrix motility genes, etc. Molecular classification of glioma with Hierarchical cluster was in accordance with pathology and revealed internal essence in tumorigenesis and development. CONCLUSION: Multiple genes play important roles in development of glioma. cDNA microarray technology is a powerful technique in screening for differentially expressed genes between two different kinds of tissues. PMID- 15130300 TI - [Microsurgical treatment of cranial-cervical type giant ependymoma within the fourth ventricle in adult patients]. AB - OBJECTIVE: To study the neuroimaging characteristics, microsurgical treatment and prognosis of cranial-cervical type giant ependymoma within the fourth ventricle in adult patients. METHODS: The clinical data of 22 patients with cranial cervical type giant ependymoma within the fourth ventricle were retrospectively analyzed. The suboccipital and C(1-3) posterior midline approach were performed in all patients. RESULTS: The tumor of all patients not only occupied the cavity of the fourth ventricle but also protruded into the cervical canal through the foramen of Magendie and extended to the cervical 2 or cervical 3 level. The tumor was totally removed in 20 patients, and subtotally removed in 2. Six patients received radiotherapy postoperatively. The time of mean follow-up of all patients was 5.5 years. One patient underwent a second operation 5 years after the first surgical procedure in other hospital due to tumor recurrence. The 5-year survival rate was 100%. All patients recovered their normal life or work. CONCLUSION: Cranial-cervical type giant ependymoma within the fourth ventricle in adult patients has a good prognosis. The surgery with postoperative adjuvant radiotherapy can obtain a optimal results. PMID- 15130301 TI - [Integrating functional magnetic resonance imaging in neuronavigation surgery of brain tumors involving motor cortex]. AB - OBJECTIVE: To assess the value of integrating blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) in neuronavigation surgery of brain tumors involving motor cortex. METHODS: A total of 58 patients with brain tumors in or directly adjacent to the motor cortex, with 18 lesions located in primary motor area, 18 lesions located in premotor area, 11 lesions located in primary motor sensory area, 9 lesions located in primary sensory area, and 2 lesions located in supplementary motor area respectively, were randomly divided into 2 groups: trial group including 30 cases undergoing BOLD navigation and control group with 28 cases undergoing routine navigation. A prospective random and matched controlled study was carried out to compare the clinical outcome between the two groups. For the patients in the trial group, the motor tasks consisted of simple flexion-extension finger movements and finger-to-thumb touching in a repeating, pre-planned sequence of either hand. A standard 1.5 T MR system had been utilized to localize the cortical motor hand area, using the BOLD contrast technique. The BOLD images were integrated with the routine navigational MR images (T1-weighted three-dimensional fast spoiled gradient recalled sequence), and then co-registered to the neuronavigation system. For the patients in the control group, the navigational MR imaging examinations were carried out only. RESULTS: The statistics analysis confirmed a good balance of main variations between the trial and control groups. The percentage of completely resection of tumors was 86.7% in trial group and 60.7% in control group (P < 0.05). The postoperative contralateral extremities muscle strength were 4.3 +/- 1.1 degree for trial group and 2.5 +/- 1.9 degree for the control group (P < 0.01). The motor functional deficit was observed in 23.3% of the cases of trial group and 71.4% of the cases in trial group (P < 0.05). The mean Karnofsky prognosis scale of the trial group was 88 +/- 27, significantly higher than that of the control group (65 +/- 32, P < 0.01). CONCLUSION: BOLD functional MR imaging is of great value in surgical planning and intraoperative functional brain mapping of motor cortex individually. To integrate BOLD data with the routine navigational MR images can supply more precise and real-time information about the relationship between lesions and neighboring cortical motor area. It should be used in neuronavigation surgery to increase the ratio of total resection of brain tumors and decrease the risk of postoperative hemiplegia. PMID- 15130302 TI - [The prevalence of deep venous thrombosis in hospitalized patients with stroke]. AB - OBJECTIVE: To investigate the prevalence and the clinical manifestations of DVT in hospitalized patients with acute stroke, and to evaluate the short-term prophylaxis effect of low molecular weight heparin (LMWH) to prevent DVT. METHODS: 488 cases with stroke admitted to the department of neurology and neurosurgery in Beijing Chaoyang Hospital between December 2001 and December 2002 were consecutively investigated. The diagnosis of stroke was identified by CT or MRI. The patients (95.5% Hans) were comprised of 328 male and 160 female, with a mean age of (65 +/- 11) years, ranging 22 approximately 93 years. The procedure of investigation: (1) A detailed history was taken and a physical examination was performed to every eligible patient. (2) The ultrasonography (US) was used for detecting both lower extremities at 7-10 days after acute stroke. (3) The ultrasound examination was repeated after a week in patients highly suspected of DVT. RESULTS: (1) The prevalence of DVT was 21.7% among the patients. (2) DVT was the most frequent (28.0%) in the patients ranging 70 approximately 79 years. (3) A higher incidence of DVT was observed in female than in male (29.4% vs 18.0%, P = 0.004). (4) The incidence of DVT in severely paralyzed patients was 25.9%. (5) Leg edema was the most common symptoms of DVT with the frequency of 13.3%, but there was a poor correlation between the symptoms and the location of DVT. And 74.5% patients were asymptomatic. (6) Most DVTs occurred in distal extremities. The DVTs in proximal veins and in distal veins were in a ratio of 1.0 to 2.3. Fibular veins were most frequently attacked (30.4%). The presence of a filling defect on ultrasound images was described in 77.3% of the DVTs. (7) A 40.0% reduction of DVT incidence was caused by LWMH treatment in ischemic stroke patients. CONCLUSIONS: (1) The hospitalized patients with acute stroke were the crowd with high risk of DVT. (2) Most DVTs were asymptomatic; the signs were nonspecific, which indicated the diagnosis of DVT couldn't depend merely on clinical manifestation. (3) Most DVTs were in distal extremities. Filling defect was the most common manifestation of ultrasonography in DVT. (4) The incidence of DVT could be reduced by using LMWH in acute ischemic stroke patients. PMID- 15130303 TI - [Comparative study of clinical characteristics and prognosis of clinically diagnosed SARS patients with positive and negative serum SARS coronavirus specific antibodies test]. AB - OBJECTIVE: To investigate clinical characteristics and long-term effects of SARS. METHODS: Clinical characteristics of 197 SARS patients in Xiao Tang-shan hospital were analyzed retrospectively, and prognosis of them were analyzed prospectively. RESULTS: Among the 197 patients, 153 patients (77.7%) have positive results of serum SARS coronavirus-specific antibodies test, and 44 patients (22.3%) have negative results. The average age of SARS and non-SARS patients were (40 +/- 12) and (31 +/- 12) years, male/female ratio were 1.0:1.6 and 2.1:1.0, average body temperature were (38.5 degrees C +/- 0.3 degrees C) and (38.1 degrees C +/- 0.4 degrees C), median of fever length were 7.0 d (0.4 approximately 50 d) and 2.3 d (0.3 approximately 37 d) respectively. The occurrence of dyspnea, malaise and gastrointestinal symptom were more often in SARS patient than in non-SARS patients. Some patients have residual symptoms (such as cough, fatigue, dyspnea, abnormality of liver function, hyperglycemia and hyperglyceridemia), and only few patients have lung fibrosis. CONCLUSION: Some patients with other respiratory diseases were misdiagnosed as SARS. There were several obvious differences of clinical characteristics between SARS patient and non-SARS patients. Prognosis of most patients were preciously well, and few still had abnormality of lung function. Residual symptoms of SARS and side effects of drugs used to treat SARS should be discriminated. PMID- 15130304 TI - [Orthotopic heart transplantation: a clinical analysis of 43 patients]. AB - OBJECTIVE: To explore the experience of orthotopic heart transplantation. METHODS: Between August 1995 and May 2003, 43 patients with end-stage heart disease, 40 males and 3 females, aged 13 approximately 58, the underlying diseases being of dilated cardiomyopathy (39 cases), hypertrophic cardiomyopathy (3 cases), and coronary cardiopathy (1 case), underwent orthotopic heart transplantation in Union Hospital, Fujian Medical University. The surgical results and relevant influencing factors were studied retrospectively. RESULTS: Seven cases died of complication within one month postoperationally, among which 5 died of acute right heart failure, 1 of septicemia, and 1 of acute renal failure, with a mortality of 16.3%. 5 cases died much later, among which 2 died of acute rejection, 2 of infection, and 1 of arrhythmia. The long-term survivors had a rather good quality of life. CONCLUSION: Heart transplantation is an effective treatment for patients with end-stage heart diseases. Appropriate selection of recipients with low pulmonary vascular resistance and proper immunosuppression treatment are key measures of successful heart transplantation. PMID- 15130305 TI - [C(1-2) posterior transarticular screw fixation (Magerl's technique) for patients with incompletely-reduced atlantoaxial dislocation]. AB - OBJECTIVE: To assess the feasibility of performing the C(1-2) transarticular screw fixation (Magerl's technique) for patients whose C(1-2) dislocation could not be reduced completely. METHODS: Nineteen consecutive patients (from 2001 Feb. to 2003 Jan.) underwent bilateral C(1-2) transarticular screw fixation, 8 of which had an incompletely-reduced atlantoaxial dislocation. The patients' clinical outcomes within- and after-operation were evaluated, and the screws position and fixation conditions were studied by postoperative CT. RESULTS: All the patients' symptoms had some improvement, no severe complications, such as injury of nerve or blood vessels, especially the vertebral arteries, were encountered. The screws were placed precisely, and purchased enough bone substance. Firm fixation obtained, no vertebral artery lesion occurred. CONCLUSION: For the patients who have atlantoaxial dislocation that could not be reduced completely, if there is enough bone to contain the screw, performing bilateral transarticular screw fixation (Magerl technique) is still feasible and safe. PMID- 15130306 TI - [Outcome and efficacy of laparoscopic splenectomy in treatment of idiopathic thrombocytopenic purpura]. AB - OBJECTIVE: To investigate the safety and efficacy of laparoscopic splenectomy (LS) in treatment of idiopathic thrombocytopenic purpura (ITP). METHODS: Twenty one patients with ITP underwent LS. The data of preoperative and postoperative platelet counts, intraoperative blood loss, recovery time, hospital stay time, and complications were analyzed prospectively. RESULTS: The platelet count was elevated rapidly in one week, and peristalsis of alimentary tract was recovered in 24 hours after operation in all cases. All patients got out of bed within 6 hours postoperatively. Their mean hospitalization time was 4 days. Complications were seen in 4 cases, of which one case underwent laparotomy, one case underwent laparoscopy once more to stop bleeding, one case developed subdiaphragmatic infection, and one case developed subcutaneous emphysema. Sixteen patients were followed up, and 2 patients relapsed. CONCLUSION: LS is a relatively safe and effective measure for treatment of ITP. PMID- 15130307 TI - [Primary culture of human periodontal ligament fibroblasts by explants with enzymatic digestion]. AB - OBJECTIVE: To increase the success rate of primary culture of human periodontal ligament fibroblasts (HPLF), and to establish an experimental model for studying HPLF in vitro. METHODS: The primary cells were isolated from human periodontal ligament by explants with enzymatic digestion method. Morphological analysis and immunocytochemical staining were used to characterize the cell lineage, and growth curve assay to evaluate the biological features of HPLF. RESULTS: The success rate of primary culture of HPLF was 77.8%. Cultured cells were spindle shaped, and had a positive reaction to antibodies against vimentin, and a negative reaction to antibodies against keratin. Their morphological and biological characteristics were similar to those of typical HPLF. Growth of HPLF obtained by this method was satisfactory. CONCLUSION: The success rate of primary culture of HPLF is significantly increased by explants combined with enzymatic digestion. The method is simple and feasible. PMID- 15130308 TI - [Detection of deletional determinants of alpha-thalassemia in Li people in Hainan province by a single tube multiplex polymerase chain reaction method]. AB - OBJECTIVE: To develop a single tube multiplex polymerase chain reaction technique (mPCR) and use it to screen 3 deletional alpha-globin genes, namely -alpha(3.7), alpha(4.2) (single gene deletion), and --SEA (South East Asia double gene deletion) in the Li people in Hainan province. METHODS: Based on previously developed 3 separate PCR methods used to detect the -alpha(3.7), -alpha(4.2), - SEA respectively, a single tube mPCR was investigated, optimized, and used to detect the genotypes of 40 samples from well-known alpha-thalassemia patients and 116 non-relative individuals of Li nationality in Hainan province. RESULTS: The results of the 40 samples detected by mPCR were the same as those by the 3 separate PCR Methods. Among the 116 sample, the combined incidence of -alpha(3.7) and -alpha(4.2) was found to be as high as 38.0%, and the -alpha(4.2) genotype was more frequent than the -alpha(3.7) genotype in the Li people. No --(SEA) deletion was found in the Li people. CONCLUSION: The mPCR technique shows a good sensitivity and specificity. The Hainan Li people has the highest incidence of alpha(3.7) and -alpha(4.2) among Chinese. PMID- 15130309 TI - [Effects of hypoxia on expression of P-glycoprotein and multidrug resistance protein in human lung adenocarcinoma A549 cell line]. AB - OBJECTIVE: To study the effects of hypoxia on expression of P-gp and multidrug resistance protein in human lung adenocarcinoma A549 cell line, and to explore the probable mechanism of hypoxia in tumor cell of MDR. METHODS: Culturing human lung adenocarcinoma A549 cell in hypoxia (2%O2) for 24 h, the expression of hypoxia inducible factor-1alpha, P-gp and multidrug resistance protein was detected by using immunohistochemistry, and after action of adriamycin or cisplatin in hypoxia (2%O2) for 24, the cell survival rate was detected by MTT. RESULTS: The expression of hypoxia inducible factor-1alpha, P-gp and multidrug resistance protein in hypoxia were higher than the expression in anoxia, and correlation between the expression of HIF-1alpha and P-gp or multidrug resistance protein was observed (P < 0.05). The resistance of adriamycin of A549 cell was enhanced in hypoxia. CONCLUSION: The resistance of tumor chemotherapy is enhanced in hypoxia. The expression of HIF-1alpha is obviously correlated with the expression of P-gp and MRP in A549 cell. PMID- 15130310 TI - [Short tandem repeat polymerase chain reaction used in prenatal diagnoses of the zygosity and common chromosomal trisomies in multiple pregnancies]. AB - OBJECTIVE: To evaluate the value of short tandem repeat polymerase chain reaction (STR-PCR) in identification of zygosity in multiple pregnancy and detection of the common chromosomal trisomies. METHODS: Amniotic fluid or fetal blood samples were collected from 38 fetuses in 17 multiple pregnant women who had indications for prenatal diagnosis or planed to be performed feticide. Of the 17 cases there were 13 sets of twins (26 fetuses) and 4 sets of triplets (12 fetuses). Parental blood samples were collected. STR-PCR technique was employed to determine the zygosity and detect trisomy 21, trisomy 18, trisomy 13, and trisomy sex chromosomes. Amniotic fluid or blood samples were collected from 18 singleton pregnant fetuses as controls for detecting chromosomal trisomies. RESULTS: All 7 cases with gestation by assisted reproductive techniques were multizygotic. In 7 cases associated with one fetal malformation, only one set of twin was dizygotic. There were no above mentioned trisomies found in the multiple pregnancies while 2 trisomy 21, one trisomy 18 and 13 respectively were detected in the singleton pregnant group. Feticide was performed on 4 multizygotic cases in which 3 were triplets and 1 was twin associated with an abnormal fetus. Following-up showed that there was no any bad effect after procedure. CONCLUSION: STR-PCR can be used in identifying the zygosity correctly and quickly while being used in detecting chromosomal trisomies. It facilitates monitoring multiple pregnancy as well as selecting appropriate methods of feticide. PMID- 15130313 TI - [Study on the inhibitory effect of small tripeptide CMS024 on tumor growth and metastasis in nude mice model of human hepatocellular carcinoma metastasis]. AB - OBJECTIVE: This experiment was designed to study the effect of small molecule tripeptide CMS024 on tumor growth and metastasis in a nude mice model of human hepatocellular carcinoma (HCC) metastasis. METHODS: Highly metastatic human HCC cell strain HCCLM6 was injected into subcutaneous region of nude mice. When the subcutaneous tumor reached 0.8 cm in diameter, the tumor was removed and cut into small pieces about 1 mm in diameter and implanted orthotopically into the liver of 20 BALB/c nude mice, which were randomized into treatment group and control group each with 10 mice. The treatment group received daily CMS024 intraperitoneal injection at the dose of 300 microg/kg beginning from the second postoperative day. The control group received intraperitoneal injection of equal amount of 0.85% sodium chloride solution. The mice were observed for signs of disease development. Thirty-five days later, the mice were sacrificed and abdominal and pulmonary metastases were recorded and peripheral blood hematological and biochemical parameters were determined. RESULTS: After 35 days of intervention, all the 10 mice in the treatment group were alive and well, but there were only 9 living mice in the control group. The mean tumor weights were 1.9 g +/- 0.5 g in the treatment group and 2.3 g +/- 0.8 g in the control group (P > 0.05). The abdominal wall metastases and intraperitoneal metastases were 100% for the control group and only 60% and 50%, respectively, for the treatment group (P < 0.05). Bloody ascites and gross intrahepatic metastatic nodules were found in 70% and 90% of nude mice, respectively, for the control group, and 20% and 40%, respectively, for the treatment group (P > 0.05). The median number of grades I, II, III and IV pulmonary metastases were 92, 24, 15 and 16, respectively, for the control group, and 24, 20, 10 and 8, respectively, for the treatment group. There were statistically significant differences in the number of grade I pulmonary metastases between the treatment group and the control group (24 vs 92, P < 0.05). No statistically significant differences in the body weight, peripheral blood hematological and biochemical parameters were observed between the two groups. No toxic effects were observed during the observation period. CONCLUSIONS: Small molecule tripeptide CMS024 demonstrated positive effects to retard tumor growth and inhibit loco-regional and long distance metastases in a nude mice model of human HCC metastasis, and no obvious toxic or side effects on the tumor bearing nude mice. PMID- 15130314 TI - [Establishment of model human stem cell of transplantation into myocardium of chick embryos]. AB - OBJECTIVE: To establish an animal model of human stem cell transplantation into myocardium in chick embryos. METHODS: Hoechsst 33,258 labeled human primordial germ cells (hPGC) were microsurgically injected into the myocardium of 633 chick embryos of 3-4 days development. Ten days after, the hearts were isolated from the 94 surviving chick embryos, embedded, and sliced. In situ hybridization (ISH) with human specific DNA Alu probe was conducted on the sections with fluorescence to detect the existence of transplanted PGC. Immunohistochemistry with human myocardium-specific antibody cTnT was conducted on the adjacent sections to observe the differentiation of human myocardial cells. RESULTS: ISH showed that PGC were detected in the myocardium of chick embryos 10 days post-operationally. Immunohistochemistry showed that the myocardium added with antibody in adjacent sections was cTnT-positive and the myocardium untreated with antibody was cTNT negative. Successful cell transplantation occurred in 15.3% +/- 2.4% of chick embryos. CONCLUSION: Establishment of an animal model of cell transplantation of human stem cells into myocardium in chick embryos is feasible. PMID- 15130315 TI - [The effect of IRESSA on H22 mouse hepatocellular carcinoma]. AB - OBJECTIVE: To investigate the effect of IRESSA (gefitinib, ZD1839) on H22 murine hepatocellular carcinoma. METHODS: Mice bearing H22 hepatocellular carcinoma were randomly divided into oral control group, Normal saline (NS) control group, cisplatin (CDDP) d1-5 group, CDDP d6-10 group, IRESSA group, IRESSA combined with CDDP early (IRESSA + CDDP d1-5) group, and IRESSA combined with CDDP lately (IRESSA + CDDP d6-10) group. IRESSA was given by daily gastrogavage for 10 days (day 1-day 10) at 100 mg/kg in body weight (BW). CDDP was given by daily intraperitoneal injection for 5 days (day 1-day 5, or day 6-day 10) at 1.2 mg/kg in BW. The growth inhibiting rate (IR) of tumor, change of BW, spleen index (SI), and amounts of blood leucocyte or hemoglobin were detected. RESULTS: IR of tumor in IRESSA group was not significantly difference with that in CDDP d1-5 group, CDDP d6-10 group, IRESSA + CDDP d1-5 group (P > 0.05). IR of tumor in IRESSA group, CDDP d1-5 group, CDDP d6-10 group, IRESSA and IRESSA + CDDP d1-5 group were 41%, 54%, 46%, and 56%, respectively. IR of tumor in IRESSA + CDDP d6-10 group (26%) was significantly lower than that in CDDP d6-10 group (P < 0.05) or in IRESSA + CDDP d1-5 group (P < 0.01). Compared with oral or NS control groups, SI and net BW in IRESSA group was not significantly difference (P > 0.05). SI and net BW in both IRESSA + CDDP d1-5 group and IRESSA + CDDP d6-10 group were lower markedly than those in IRESSA group (P < 0.01). CONCLUSION: Tumor growth of H22 bearing mice was markedly inhibited by IRESSA. PMID- 15130316 TI - [The estimation of protective efficacy of the fusion gene vaccine encoding tubercle antigen 85B and MPT64 in mice challenged with Mycobacterium tuberculosis]. AB - OBJECTIVE: To evaluate the protective efficacy of the fusion DNA vaccine (AM) encoding tubercle Ag85B and MPT64 in mice infected with Mycobacterium tuberculosis. METHODS: C57BL/6 mice were intramuscularly immunized with the DNA vaccines. The mice were challenged with 10(6) CFU H37Rv via lateral tail vein 35 days later after the third immunization for DNA vaccine groups and 100 days later for BCG vaccinated group. The mice in vaccinated groups and control groups were sacrificed 42 days later following challenge. The lungs and spleens were removed respectively, and the number of CFU in organs and histopathologic changes was determined. The antibody level, IFN-gamma, IL-4 and the survival time in all of the mice were evaluated. RESULTS: Antibody titer of pcDNA/Ag85B + pcDNA/MPT64 group and pcDNA/AM group was higher than that of other groups (P < 0.05). The level of IFN-gamma produced by spleen lymphocytes and spleen lymphocyte proliferation from BCG group, pcDNA/Ag85B, pcDNA/Ag85B + pcDNA/MPT64 group and pcDNA/AM group was higher than that of other groups (P < 0.05). No IL-4 was found in all groups. The number of bacterial colonies in the lungs and spleens was significantly decreased at 6th week postchallenge in all the vaccinated groups (P < 0.05), especially in BCG group (P < 0.01). The pulmonary histopathological changes were observed 6 weeks later following challenge with M. tuberculosis H37Rv. In PBS and pcDNA3.1 groups, the lesion was characterized by seroplastic inflammatory infiltration and lung tissue necrosis, in BCG group by granulomas and numerous macrophages, lymphocytes and a few epithelioid cells. The lesion in pcDNA/Ag85B groups was characterized by seroplastic inflammatory infiltration and a few macrophages, in pcDNA/Ag85B + pcDNA/MPT64 group and pcDNA/AM group, by granulomas, numerous macrophages and lymphocytes. The lesion in spleen was different from the lung and characterized by proliferative lymphocytes and inflammatory infiltration. The results in spleen were similar to those in lung. The survival time of BCG vaccinated mice after challenge with M. tuberculosis H37Rv was longer than that of other groups. The survival time of AM group was longer than that of other DNA vaccine groups. CONCLUSION: The pcDNA/AM can improve the protective efficacy in immunized mice against M. tuberculosis. PMID- 15130322 TI - [Prognostic analysis of lung function and chest X-ray changes of 258 patients with severe acute respiratory syndrome in rehabilitation after discharge]. AB - OBJECTIVE: To analyze the clinical characteristics and prognostic changes of rehabilitating severe acute respiratory syndrome (SARS) patients through regular lung function tests and lung imaging studies after discharge and to retrospectively analyze the treatment data of these patients. METHODS: 258 discharged SARS patients received regular SARS-Co virus IgG test, lung function test and chest X-ray and/or high resolution computerized tomography (HRCT) examination at General Hospital of PLA two months after discharge, and the treatment data of these patients were retrospectively analyzed. RESULTS: 80.6% patients (208 of 258 patients) were positive for SARS-Co virus IgG. 21.3% patients (55 of 258 patients) showed lung diffusion abnormity (D(LCO) < 80%pred). Compared to 155 SARS-Co virus IgG positive patients without lung diffusion abnormity and 50 SARS-Co virus IgG negative patients, the 53 SARS-Co virus IgG positive patients with lung diffusion abnormity had longer fever course, higher dosages of glucocorticoid therapy, higher percentage of oxygen therapy and non invasive ventilation. 51 of the 53 patients with lung diffusion abnormity received lung function test after one month, and the results of D(LCO) improved in 80.4% patients (41 of 51 patients). 40 of 51 patients with lung diffusion abnormity showed lung fibrosis, and the fibrosis decreased in 55% patients (22 of 40 patients) after one month. CONCLUSIONS: This finding suggests that lung fibrosis caused by SARS mostly occurs in severe patients, and it can resolve spontaneously. D(LCO) may be more sensitive than HRCT in evaluating the fibrotic changes. PMID- 15130323 TI - [Covert infection of severe acute respiratory syndrome in health-care professionals and its relation to the workload and the type of work]. AB - OBJECTIVE: To study severe acute respiratory syndrome (SARS) covert infection in health-care professionals and its relation to work intensity and type of work. METHODS: Specific antibodies to SARS virus in 1 127 serum specimens from health care professionals who had a close contact history with patients of SARS and 92 healthy persons were assayed with ELISA. RESULTS: The health-care professionals consisted of 287 men and 840 women, aged 21 - 55 years (mean 34 +/- 9 years). The overall positive rate of specific IgG and IgM was 2.57% (29/1 127); the positive rates in nurses, doctors and laboratory workers, and supervisors were 3.46%, 1.41%, 2.86%, respectively. 34.5% of the infected worked in the ICU, and 65.5% in the general wards. None of them presented clinical SARS. Medical examination was also carried out for 313 people who had close contact with the 29 antibody positive health care professionals, but no clinical SARS was found. CONCLUSIONS: Covert infection can be found in the health-care professionals and it showed no contagiousness. The positive rate of specific antibodies was higher in ICU where the work intensity was higher. The positive rate in nurses was higher than in doctors. PMID- 15130324 TI - [Resistance surveillance of common community respiratory pathogens isolated in China, 2002 - 2003]. AB - OBJECTIVE: To investigate antimicrobial resistance of common community respiratory pathogens isolated in China, 2002 - 2003. METHODS: 779 strains of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Group A beta-haemolytic Streptococci and oxacillin-susceptible Staphylococcus aureus (MSSA) were isolated from patients with community-acquired respiratory tract infections at 5 hospitals in China from April 2002 to 2003. Meanwhile, 185 strains of S. pneumoniae, H. influenzae and M. catarrhalis were isolated from nasopharynx swabs at 2 day-care centers in Beijing. Agar dilution method was used to determine the minimal inhibitory concentration (MICs) of cefprozil and other 9 antibiotics against these strains. RESULTS: The prevalence of penicillin intermediate S. pneumoniae (PISP) was 23.9% and that of penicillin-resistant S. pneumoniae (PRSP) was 22.7% at 5 cities in China. The prevalence of PISP were 44.1% in Hangzhou, 26.2% in Wuhan, 21.5% in Shenyang, 20.8% in Shanghai, 18.5% in Beijing, and 12.7% at day-care centers in Beijing;the prevalance of PRSP were 34.9% in day-care centers, 31.9% in Shanghai, 27.9% in Wuhan, 22.1% in Hangzhou, 13.8% in Shenyang and 8.6% in Beijing. The susceptible rate of levofloxacin in S. pneumoniae was 96.3%. 9.5% of H. influenzae and 87.4% of M. catarrhalis produced beta-lactamases. The susceptibility of amoxicillin/clavulanate, cefaclor, cefprozil, cefuroxime, ceftriaxone, azithromycin, and levofloxain in these two species ranged from 96.4% to 100%. The resistance rate of azithromycin in S. pneumoniae was higher than 60%. Cefprozil MICs against PISP, Group A beta haemolytic Streptococci and MSSA were 4 - 16 fold lower than cefaclor. CONCLUSIONS: Antimicrobial resistance in respiratory pathogens, especially S. pneumoniae is increasing. It brings concerns that high macrolide resistance was found in gram-positive cocci. Cefprozil was more active than cefaclor against respiratory pathogens. PMID- 15130325 TI - [Alveolar damage of pneumocystis carinii pneumonia in renal transplantation recipients]. AB - OBJECTIVE: To study the alveolar ultrastructural changes and the interaction between Pneumocystis carinii (PC) and alveoli in Pneumocystis carinii pneumonia (PCP) patients after renal transplantation. METHODS: Twenty-seven patients with suspected PCP after renal transplantation were examinated by bronchoscopy with bronchoalveolar lavage (BAL) and transbronchial lung biopsy (TBLB). The BAL fluid was centrifuged and the sediments were stained for PC. Cases for which electron microscope showed alveolar tissue in TBLB specimen were included. Then the clinical features, PC, alveolar epithelial damage, exudate in alveolar space, inflammatory cell infiltration, and fibrous tissue in the interstitial space were analyzed and evaluated. RESULTS: Twenty-three cases were studied. The mean time from renal transplantation to onset of illness was 5.6 months, and that from onset of illness to hospitalization was 5.5 days. Clinical features included fever, dyspnea, unproductive cough, and scanty chest signs, to hypoxemic respiratory failure. Chest CT showed diffuse lung interstitial changes in 22 of the 23 cases, 9 with consolidation. After treatment with SMZco, the fever resolved in 1 - 5 days, and the general state of the patients became better, and 19 patients were dischaged within 1 month. PC in BAL fluid was found by special staining in 18 patients, while PC was found by electron microscope in 14 patients. In most cases PC was few in the lung tissue, but in 3 cases abundant PC filled the alveolar space. PC was seen in two forms, the cyst and the trophozoite. Electron and light microscopes showed alveolar exudate, inflammation in interstitium and alveolar space, interstitial fibrosis, and alveolar epithelial damage in all patients. CONCLUSION: In PCP patients after renal transplantation there was marked alveolar damage, which was the major pathological change in the lung. PMID- 15130326 TI - [The experimental study on the inhibitory effect of tripterine on airway inflammation in asthmatic mice]. AB - OBJECTIVE: To investigate the effect and mechanism of tripterine on airway inflammation in asthmatic mice. METHODS: 30 BALB/c mice were randomly divided into a control group, an asthmatic group (ovalbumin-sensitized) and a tripterine (1 mg/kg, introperitoneally)-treated group. Pathologic changes in lung tissues, number of eosinophils in bronchoalveolar lavage fluid (BALF) and expression of stem cell factor (SCF) protein in lung were observed. In vitro, we established coculture system of bone marrow derived-mast cells from C57B6 mice and fibroblast NIH3T3, which were then treated by 2 micro mol/L of tripterine, and compared with NIH3T3 and mast cells. Concentration of histamine and eotaxin in supernants of coculture was measured by fluorometry and ELISA respectively, and expression of SCF protein in fibroblasts from cocultures was analyzed with immunohistochemistry. RESULTS: Compared with the asthmatic group, less inflammatory cell infiltration in lung tissues was observed in the tripterine treated group. There was significant difference in the number of eosinophils in BALF between the tripterine-treated group [(0.56 +/- 0.03) x 10(6)/L] and the asthmatic group [(1.25 +/- 0.40) x 10(6)/L, P < 0.05]. So was the expression of SCF protein in lung tissue [0.74 +/- 0.20, 2.50 +/- 0.19, P < 0.01]. In vitro, the concentration of histamine and eotaxin in coculture supernants and the expression of SCF protein in fibroblasts from coculture were (3.83 +/- 0.41) ng/ml, (5.79 +/- 0.40) ng/ml and (95 +/- 3)%, respectively; after tripterine intervention, the data changed to (2.88 +/- 0.35) ng/ml, (4.24 +/- 0.29) ng/ml, (17 +/- 5)% (all P < 0.01). CONCLUSIONS: Tripterine might suppress airway inflammation in asthmatic mice, probably by downregulating the expression of SCF in fibroblasts, then inhibiting the production of histamine and eotaxin in mast cells. PMID- 15130327 TI - [Influence of peroxisome proliferator-activated receptor gamma on airway inflammation of guinea pigs with asthma]. AB - OBJECTIVE: To investigate the influence and mechanism of peroxisome proliferator activated receptor gamma (PPAR-gamma) and its ligand agonist rosiglitazone on airway inflammation of guinea pigs with asthma. METHODS: 35 guinea pigs were divided into 5 groups by random number meter: a control group (A group), an asthma group (B group), a dexamethasone (DXM) group (C group), a rosiglitazone group (D group), and a rosiglitazone + 1/2 DXM group (E group), with 7 guinea pigs in each. Bronchoalveolar lavage (BAL) cell count and differential was studied, and the pathologic alteration of the bronchi and the lung tissue was observed. The expression of PPAR-gamma, COX-2 was measured by RT-PCR. RESULTS: BAL eosinophil count was 0.050 +/- 0.020 in D group, 0.110 +/- 0.020 in B group, the difference being significantly (t = 5.61, P < 0.001). The total cell number and neutrophils were (15.5 +/- 3.9) x 10(8)/L and 0.069 +/- 0.020 in D group, and (19.9 +/- 4.3) x 10(8)/L and 0.076 +/- 0.020 in B group, the difference being not significant (t = 2.02, 0.66, P > 0.05 respectively). The thickness of airway wall of D group was (22.0 +/- 5.0) micro m, and in B group it was (28.0 +/- 5.0) micro m, the difference being significant (t = 2.61, P < 0.05), but the thickness of mucosa and submucosa of D group (12.2 +/- 2.9) micro m was not different as compared with B group (14.9 +/- 3.3) micro m (t = 1.63, P > 0.05). Expression of PPAR-gamma mRNA of D group 19.5 +/- 3.0 was not different compared with B group 18.1 +/- 3.1 and A group 15.6 +/- 2.9 respectively (t = 0.92, 0.49, P > 0.05, respectively). When the expression of COX-2 mRNA of B group 49 +/- 7 was compared with D group 39 +/- 6, the difference was significant (t = 2.77, P < 0.05). CONCLUSIONS: Rosiglitazone decreases airway eosinophils and the thickness of airway wall in guinea pigs with asthma, via suppression of COX-2 mRNA expression by activating PPAR-gamma. The anti-inflammatory effect of PPAR-gamma may be associated with the use of ligand agonist and(or) glucocorticoids. PMID- 15130328 TI - [Hypoxia-inducible factor 1alpha regulates vascular endothelial growth factor's roles on pulmonary arteries of rats with hypoxia-induced pulmonary hypertension]. AB - OBJECTIVE: To investigate the dynamic expression of hypoxia inducible factor 1alpha (HIF-1alpha) and vascular endothelial growth factor (VEGF) in pulmonary arteries of rats with hypoxia-induced pulmonary hypertension. METHODS: Forty male adult Wistar rats were randomly divided into five groups: a control group (C group) and groups with hypoxia for 3, 7, 14 and 21 days (H(3), H(7), H(14) and H(21) group), eight rats per group. Mean pulmonary pressure (mPAP), vessel morphometry and right ventricle hypertrophy index (RVHI) were measured. Lungs were inflation fixed for in situ hybridization and immunohistochemistry. RESULTS: mPAP increased significantly after 7-day of hypoxia [(18.41 +/- 0.37) mm Hg, P < 0.05], reaching its peak after 14-day of hypoxia, then remained on the high level. Pulmonary artery remodeling index (outer diameter 100 - 150 micro m) and RVHI became evident after 14-day of hypoxia. Expression of HIF-1alpha protein in control group was poorly positive, but was up-regulated in pulmonary arterial tunica intima of all hypoxic rats. In pulmonary arterial tunica media, the levels of HIF-1alpha protein was markedly up-regulated after 3-day (0.178 +/- 0.017, P < 0.05), reaching its peak after 7-day of hypoxia (0.221 +/- 0.021, P < 0.05), then tended to decline after 14-day and 21-day of hypoxia. HIF-1alpha mRNA staining was poorly positive in control, hypoxia for 3 days and hypoxia for 7 days, but began to increase significantly after 14-day of hypoxia (0.203 +/- 0.024, P < 0.05), then remained stable. Expression of VEGF protein began to increase after 7 day of hypoxia (0.074 +/- 0.022, P < 0.05), reaching its peak after 14-day of hypoxia (0.147 +/- 0.017, P < 0.05), then remained on the high level. VEGF mRNA in control and hypoxia for 3 days was poorly positive, but began to increase after 7-day of hypoxia (0.138 +/- 0.010, P < 0.05), and remained on the high level thereafter. VEGF mRNA located mainly in tunica intima and tunica media, whereas VEGF protein located predominantly in tunica intima. Linear correlation analysis showed that HIF-1alpha mRNA, VEGF, and mPAP were correlated with vessel morphometry and RVHI (P < 0.01). HIF-1alpha protein (tunica intima) was positively correlated with VEGF mRNA and protein (P < 0.01). CONCLUSIONS: HIF 1alpha and VEGF are both involved in the pathogenesis of hypoxia-induced pulmonary hypertension in rats. HIF-1alpha protein may regulate the expression of VEGF gene by transcriptional activation, resulting in the occurrence and development of hypoxic pulmonary hypertension. PMID- 15130329 TI - [Detection of KatG 315 gene mutation associated with isoniazid resistance in Mycobacterium tuberculosis by template-directed dye-terminator incorporation with fluorescence polarization detection technology]. AB - OBJECTIVE: To study the relationship between KatG 315 mutation and isoniazid (INH) resistance in M. tuberculosis by template-directed dye-terminator incorporation with fluorescence polarization detection (TDI-FP), and to develop a new method that can detect INH-resistant M. tuberculosis precisely and quickly. METHODS: Isolates of M. tuberculosis resistant to INH from 82 tuberculosis patients were cultured on culture medium with different INH concentrations. DNA of all M. tuberculosis samples was extracted and amplification of 271 bp KatG gene fragment was carried out by polymerase chain reaction (PCR). After digestion of the excess primers and dNTPs in PCR products by clean-up reagents, template directed dye-terminator incorporation reaction was performed, and the Acyclo terminators labeled Rhodamine 110 (R110) or 6-carboxytetramethylrhodamine (TAMRA) were incorporated into the detection primers specifically. Then fluorescence polarization value was measured using Victor2 multi-label counter and the genotypes of the mutation site in 315 condon of KatG gene of all samples were investigated. RESULTS: There were 15 out of the 29 isolates of M. tuberculosis with high INH-resistance showed G-->C mutation in 315 condon of KatG gene, including 4 isolates with both mutated and non-mutated trains, the mutation frequency being 52%. In 32 isolates of M. tuberculosis with low INH-resistance, 15 were mixed infection, the mutation frequency being 47%. No mutation in 315 condon of KatG gene was found in 21 susceptible isolates of M. tuberculosis. CONCLUSIONS: The G-->C mutation in 315 condon of KatG gene was associated with INH-resistance of M. tuberculosis. TDI-FP technology was a reliable, easy to use and high-through method to detect mutation of KatG gene. It may be a useful technique for the diagnosis of INH-resistance in the future. PMID- 15130330 TI - [Study on gene knock-out in Mycobacterium BCG]. AB - OBJECTIVE: To establish the methodology of plasmid for gene knock-out in Mycobacterium BCG. METHODS: We designed two pairs of primers for amplification of MDP1 gene and inserted two fragments into pKO plasmid, and then the recombinant plasmid for MDP1 gene knock-out was obtained, and named pKO-MDP1. Gene exchange took place within the genome of BCG after pKO-MDP1 plasmid was transformed into Mycobacterium BCG. The strain of Mycobacterium BCG with MDP1 gene knock-out was selected, and the curve of growth rate was studied. RESULTS: The target strain was that of the positive strain by two step PCR and one step sucrose counter selection, without growth in culture media with hygomycin. The value of A(600) per 12 hours was detected for sixteen days. A "S" shaped growth curve was detected. However, there was no significant difference in the growth rates between the wild type Mycobacterium BCG strain and the gene knock-out Mycobacterium BCG strain. CONCLUSIONS: The plasmid of pKO was a useful tool for gene knock-out in Mycobacteria. MDP1 maybe one of the factors influencing the growth rate, but it was not the only one. PMID- 15130331 TI - [Molecular cloning, purification, and serological characterization of four specific antigens of Mycobacterium tuberculosis]. AB - OBJECTIVE: To express the 14,000, 37,000, and 6,000 early secretory antigenic target (ESAT-6) and mtb81 antigen genes in bacteria, and to purify the product and determine their activity. METHODS: The 14,000, 37,000 , ESAT-6, and mtb81 antigen genes were amplified from Mycobacterium tuberculosis genomic DNA by polymerase chain reactions and cloned into pGEX 4T-1 expression vector. BL21 strain of Escherichia coli was transformed with the recombinant vectors and induced to express recombinant proteins. The proteins were purified by affinity chromatography. The biological activity of purified proteins were estimated by enzyme-linked immunoabsorbant assay (ELISA). RESULTS: The BL21 strains of Escherichia coli with recombinant vectors showed high level of 14,000, 37,000, ESAT-6, and mtb81 gene expressions after induction. The products were purified successfully and showed high antigenicity and specificity. The sensitivity of 38,000, 14,000, ESAT-6, and mtb81 were 54%, 60%, 44%, and 36%, respectively. CONCLUSION: The expressions and purifications of recombinant 14,000, 37,000, ESAT 6, and mtb81 antigens with natural activity facilitate their research and application. PMID- 15130332 TI - [The effect of Na(+)/H(+) exchanger-1 (NHE-1) antisense expression vector on NHE 1 gene expression in human lung adenocarcinoma cells and its biological significance]. AB - OBJECTIVE: To explore the inhibitory effect of Na(+)/H(+) exchanger-1 (NHE-1) antisense expression vector on NHE-1 gene in human lung neoplasm cells and to observe its biological effect. METHODS: NHE-1 antisense vector (pNHE-1) was introduced into human lung adenocarcinoma cell line A549 cells with cationic liposome transfection methods. Semi-quantification RT-PCR was performed to analyze the expression level of NHE-1 mRNA in A549 cells and transfected A549 cells. Intracellular pH (pHi) values were measured with fluorescence spectrophotometer. The growth characteristics of the cells was observed. Tumor cell apoptosis was observed with in situ apoptosis assessment. RESULTS: PCR verified the integration of antisense vector with the genomic DNA of transfected cells in positive clones. Semi-quantification RT-PCR showed that the NHE-1 gene expression level was significantly lower in transfected A549 cells (0.42 +/- 0.06) than in those untransfected (0.71 +/- 0.08, P < 0.01) and those transfected with pLXSN (0.69 +/- 0.16, P < 0.01). Compared with A549 cells, pHi values decreased significantly in transfected A549 cells at 12 h, 24 h and 48 h (6.990 +/- 0.005, 6.840 +/- 0.005 and 6.750 +/- 0.005 respectively vs 7.150 +/- 0.004, 7.140 +/- 0.007 and 7.120 +/- 0.008 respectively, P < 0.001). The growth of transfected A549 cells was significantly slower (P < 0.01), the double time was longer (3.20 vs 4.97) and the apoptosis rate was markedly higher [(24.50 +/- 3.62)% vs (1.50 +/- 0.55)%, P < 0.01]. CONCLUSION: The antisense vector was successfully transfected to A549 cells with liposome and integrated with transfected cell genome, and the expression of NHE-1 mRNA of A549 cells was inhibited, thereby resulted in a decrease of the pHi value by means of reduction of Na(+)/H(+) exchanging activity, with the inhibition of proliferation and growth of A549 cells and increased of A549 cell apoptosis. PMID- 15130334 TI - [Diagnosis and treatment of pulmonary fungal infection in intensive care unit]. PMID- 15130335 TI - [Advance in the study of genes associated with susceptibility of tuberculosis]. PMID- 15130343 TI - [Clinical characteristics of 1241 cases of intrahepatic cholestasis of pregnancy]. AB - OBJECTIVE: To investigate the clinical characteristics of intrahepatic cholestasis of pregnancy (ICP). METHODS: Totally 1241 cases of ICP during the 10 years from Jan 1991 to Dec 2000 in our hospital were retrospectively reviewed. RESULTS: Forty-four (3.5%) patients were multiple gestation. One hundred and one (8.1%) patients had pregnancy induced hypertension. ICP recurred in 30.2% multipara (38/126). On average, it occurred at gestational week of 32.6. Pruritus was the first symptom in 1201 (96.8%) patients while 8 (0.6%) patients had jaundice and 28 (2.3%) patients had abnormal liver function at the onset. Serum total bile acid (TBA), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were usually mild or moderately elevated, while some patients had normal TBA (17.7%), ALT (15.6%), or AST (17.1%) level. The albumin/globulin ratio was reversed in 35 (3.2%) patients. CONCLUSION: ICP tends to recur in subsequent pregnancy. It is more likely to develop in multiple gestation. Pruritus is the most common and prominent manifestation. Elevated TBA and ALT or AST level is helpful for diagnosis. It should be noted that the above manifestations might not be typical in some patients. PMID- 15130344 TI - [Study of the effects of utero-placental ischemia on the body and nervous system development in fetal rats]. AB - OBJECTIVE: To discuss the effects of utero-placental ischemia on the body and nervous system development in fetal rats. METHODS: By clamping the unilateral uterine artery of the rat, we produced a utero-placental ischemia model. The opposite uterus of the rat with normal uterine artery supply served as control. We compared the body weight, weight of brain, and the expression of growth associated protein-43 (GAP-43) mRNA in cerebral tissue by RT-PCR in the 13 day (group 1) and 17 day (group 2) old fetal rats respectively. RESULTS: The body weight and weight of brain in group 1 were 3.2 g and 0.16 g respectively, significantly lower than those of control 1 of 3.6 g and 0.18 g respectively (P < 0.01, P < 0.05). The body weight and weight of brain in group 2 were 3.1 g and 0.16 g respectively, also significantly lower than those of control 2 of 3.72 g and 0.17 g respectively (P < 0.01, P < 0.05). The expression of GAP-43 mRNA in cerebral tissue of the group 1 (1.10) and its control (1.12) showed no obvious difference (P > 0.05). However, GAP-43 mRNA in cerebral tissue of the group 2 (1.06) was significantly decreased compared with that of its control (1.21) (P < 0.01). CONCLUSION: Clamping the uterine artery can mimic intrauterine ischemia and produce FGR model. Intrauterine ischemia in the late stage of pregnancy may decrease the expression of GAP-43 mRNA, which might affect the development of nervous system in rats. PMID- 15130345 TI - [Study on the mechanism of intrauterine infection of hepatitis B virus]. AB - OBJECTIVE: To study the possible mechanism of intrauterine infection of hepatitis B virus (HBV). METHODS: HBV DNA was examined in amniotic fluid, and vaginal secretion of 59 HBsAg positive mothers and in cord blood of their neonates by PCR. Ten negative hepatitis B virus marker (HBVM) mothers and their neonates were served as control. HBsAg and HBcAg in placenta were examined by avidin biotin complex (ABC) method. RESULTS: The detection rate of HBV DNA in amniotic fluid, vaginal secretion and neonatal cord blood of the study group were 47.5% (28/59), 52.5% (31/59) and 45.8% (27/59) respectively. HBsAg and HBcAg in placenta was distributed in the following descending order: maternal decidual cells, trophoblastic cells, villous mesenchymal cells and villous capillary endothelial cells. But the distribution was in reverse order in 4 placentas. HBsAg and HBcAg were detected in amniotic epithelial cells in 32 mothers. CONCLUSION: The main route of HBV transmission from mother to fetus is transplacental, from maternal side of placenta to fetal side. However, HBV intrauterine infection may take place through other routes. PMID- 15130346 TI - [Clinical characteristics and pathologic study of placental chorioangioma]. AB - OBJECTIVE: To describe the clinical characteristics and gross picture of placental chorioangioma. METHODS: Clinicopathologic data from twelve cases of placental chorioangioma were retrospectively studied. RESULTS: (1) In twelve cases pathologically diagnosed as placental chorioangioma, five cases were consistent with the clinical diagnosis; two cases were misdiagnosed as placental abruption; one case was misdiagnosed as lipoma; one case was misdiagnosed as placental cyst; and three cases were missed. Color Doppler ultrasound examination was performed in four cases ante partum, in which two cases were diagnosed correctly and two cases were missed. (2) Premature labor occurred in three cases of chorioangioma, two of which were associated with polyhydramnios. Two were complicated with pregnancy-induced hypertension syndrome. Fetal death occurred in two cases and fetal abnormality in one case and neonatal asphyxia in four cases. (3) Placental chorioangiomas of all cases ranged from 2 cm x 2 cm x 2 cm to 11 cm x 8 cm x 8 cm in size. There was single tumor in nine cases and two tumors in three cases. The tumor appeared deep red in six cases, grayish red in three cases, pale yellow in one case, partial deep red and partial gray white in one case, partial deep red and partial pale yellow in one case. CONCLUSIONS: Placental chorioangioma can induce several complications such as premature labor, fetal death, polyhydramnios and neonatal asphyxia. Color Doppler is helpful to differential diagnosis of placental masses particularly placental chorioangioma, though the later is difficult to diagnose ante partum. Placental chorioangioma is varied in size with deep red in appearance for most of them and maybe yellow or gray white due to mixed with other tissues or lack of blood supply. PMID- 15130347 TI - [Effect of Bushenyiqihexue recipe on the expression of endometrial pinopodes in blastocyst implantation dysfunctional mice]. AB - OBJECTIVE: To observe the effect of Bushenyiqihexue recipe on the expression of endometrial pinopodes in blastocyst implantation dysfunctional mice. METHODS: To induce blastocyst implantation dysfunctional mice model, Kunming mice were injected subcutaneously with mifepristone on day 4 of pregnancy. First, Kunming mice were randomly assigned to the control group, the model group and treated group postcoitally. Every morning from day 1 of pregnancy (Pd1), the treated group was administered the Bushenyiqihexue recipe decoction, while the model group and control group were administered saline orally. On Pd4, the model and treated groups were injected subcutaneously with mifepristone diluted in propylene glycol, while the control group was injected with saline only. The expression of endometrial pinopodes between 21:30 and 22:00 on Pd4 (T1) and between 9:30 and 10:00 on Pd5 (T2) was determined by scanning electron microscopy. RESULTS: Compared with control group (90%, 14.7 +/- 1.4), pregnancy rate (25%) and mean implanted blastocysts (7.7 +/- 1.3) in model group were remarkably reduced (P < 0.05, P < 0.001). Compared with the model group, pregnancy rate (55%) and mean implanted blastocysts (12.3 +/- 0.8) in treated group were significantly improved (P < 0.05, P < 0.01). At T1, abundant developing pinopodes were expressed in endometrial surface in control group, whereas abundant fully developed pinopodes expressed at T2. At T1, only a few pinopodes were expressed locally in endometrial surface in model group, while completely disappeared at T2. At T1, endometrial surface in treated group expressed a lot of developing pinopodes which lagged behind slightly that of the control group in time, whereas many fully developed pinopodes expressed at T2. CONCLUSIONS: Decreased pinopodes expression at T1 and complete loss expression of pinopodes at T2 seem to be the reasons for blastocyst implantation dysfunction induced by mifepristone. It is suggested that Bushenyiqihexue recipe could improve pinopodes expression in endometrial surface, eventually better uterine receptivity and improve the blastocyst implantation. PMID- 15130348 TI - [Inducing peripheral materno-fetal immuno-tolerance by blockade of costimulatory signal at early stage of gestation of the abortion-prone murine model]. AB - OBJECTIVE: To study effect of blockade of costimulatory signal CD(80), CD(86) at the early stage of gestation on the maternal peripheral immuno-tolerance status to paternal antigen and pregnant outcome of murine abortion-prone model. METHODS: The experiments were performed in two groups, using CBA/J x BALB/c matings as the normal pregnancy model and CBA/J x DBA/2 matings as the abortion-prone model. The pregnant CBA/J mice which had mated with DBA/2 or BALB/c male mice respectively were injected intraperitoneally with purified rat isotype IgG or purified rat anti-mouse CD(80), CD(86) mAb at day 4 of gestation (time of implantation). The proliferation response of maternal splenic immuno-competent cells to paternal splenic cells as stimulator was analyzed by one way mixed lymphocyte reaction, and IL-2 production was assayed by ELISA to evaluate the immuno-tolerance of maternal peripheral immuno-competent cells to paternal antigen at day 9 of gestation. The embryo resorption rate was counted at day 14 of gestation. RESULTS: In the CBA/J x DBA/2 matings, anti-CD(80), CD(86) mAb administered at day 4 of gestation reduced significantly the embryo resorption rate, and induced a significantly lower proliferation response and IL-2 production by maternal splenic immuno-competent cells to paternal antigen compared with rat isotype IgG. CONCLUSIONS: In vivo blockade of costimulatory signal at day 4 of gestation can induce the immuno-tolerance of maternal peripheral immuno-competent cells to paternal antigen, thus leading the embryo resorption rate of the abortion-prone model to that of normal pregnancy model. PMID- 15130349 TI - [C677T and A1298C mutation of the methylenetetrahydrofolate reductase gene in unexplained recurrent spontaneous abortion]. AB - OBJECTIVE: To investigate the association between the C677T and A1298C mutation of 5,10-methylenetetrahydrofolate reductase (MTHFR) gene and unexplained recurrent spontaneous abortion (URSA) in Chinese population. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the mutation of C677T and A1298C of MTHFR in 148 cases with URSA and 82 normal controls. RESULTS: (1) The distribution frequencies of C667T associated 3 genotypes between the URSA and control group showed statistically significant difference (P = 0.012). The frequencies of C677T genotypes were: CC (33.3%), CT (53.1%), TT (13.6%) in URSA group and CC (52.4%), CT (51.5%), TT (6.1%) in control group, respectively. And the frequency of CC genotype in URSA group was decreased significantly (P = 0.005), while the frequency of T allele in URSA was increased (P < 0.005). (2) The prevalence of the MTHFR A1298C associated 3 genotypes and A/C alleles in URSA group did not differ significantly from the control. (3) According to the linkage analysis of C677T and A1298C, 8 linkage genotypes were found, and the frequency of 677CC/1298AA in URSA was significantly lower compared with the control, the linkage of 677 (CT + TT)/1298CC was only observed in URSA group. CONCLUSIONS: The mutations of MTHFR C677T and A1298C play a role in the mechanism of unexplained recurrent spontaneous abortion. PMID- 15130350 TI - [Insulin-like growth factor II and its receptor gene expression in the endometrium of women with unexplained infertility]. AB - OBJECTIVE: To investigate the expression of insulin-like growth factor II (IGF II) and its type I receptor (IGF-IR) and type II receptor (IGF-IIR) in the endometrium of women with unexplained infertility and their relation to steroid levels. METHODS: In situ hybridization method was utilized to detect the location of IGF-II, IGF-IR and IGF-IIR mRNA. Reverse transcription-polymerase chain reaction (RT-PCR) method was employed to detect quantitatively IGF-II, IGF-IR, and IGF-IIR mRNA expression levels in the endometrium during mid-luteal phase. Serum concentrations of estradiol and progesterone were measured by radioimmunoassay. Thirty-eight patients with unexplained infertility were included as study group, and 20 patients with men factor infertility or normal volunteer women were selected as control group. RESULTS: IGF-II and IGF-IIR mRNA were diffusely distributed in the cytoplasm of stromal cells and IGF-IR mRNA was observed in the cytoplasm of epithelial cells. Endometrial IGF-II, IGF-IR and IGF IIR mRNA expression during mid-luteal phase in study group were significantly lower than those in the control group respectively (0.71 +/- 0.34 vs 0.96 +/- 0.34, P < 0.05; 0.62 +/- 0.28 vs 0.93 +/- 0.51, P < 0.05; 0.49 +/- 0.27 vs 0.73 +/- 0.36, P < 0.05). The serum progesterone level of study group was significantly lower than that of control group, (34 +/- 15) nmol/L vs (53 +/- 17) nmol/L (P < 0.05). The levels of IGF-II mRNA were positively correlated with IGF IR mRNA and IGF-IIR mRNA in both groups. There were significant positive correlations between serum progesterone level and IGF-II, IGF-IR and IGF-IIR mRNA. CONCLUSIONS: IGF-II and its receptors mRNA play important roles in the process of implantation. Their decreased expression in relation with lower progesterone level may be one of the major causes of unexplained infertility. PMID- 15130351 TI - [Temporal and spatial expression of estrogen activity-related molecules in eutopic endometrium of adenomyosis]. AB - OBJECTIVE: The aim of this study was to demonstrate the temporal and spatial expression of estrogen receptor (ER), 17beta-hydroxysteroid dehydrogenase (17beta HSD) in uterine endometria and the endometrial-myometrial interface (EMI) of adenomyosis and the effects of estrogen activity-related molecules on the occurrence of adenomyosis. METHODS: Thirty-three cases of normal endometria (and myometrial) and eighteen cases of endometria (and myometrial) with adenomyosis were collected. Immunohistochemical assay was performed to locate the ERalpha, ERbeta, 17beta-HSDI and 17beta-HSDII in endometria and EMI. RESULTS: The ERalpha positive cell number in glandular epithelial cells at the early proliferative phase increased evidently in adenomyosis (90%), while it was 60% in normal endometria. We found ERalpha signal in cytoplasm in glandular epithelial cells of adenomyosis endometria as well as in nucleus. Compared with normal endometrium (early proliferative phase: +; late proliferative phase: ++; late secretory phase: +), eutopic endometrium with adenomyosis exhibited a higher level of 17beta-HSDI (early proliferative phase: ++; late proliferative phase: +++; late secretory phase: ++). The intensity of 17beta-HSDIIin glandular epithelial cells of eutopic endometrium with adenomyosis (early proliferative phase: +++; secretory phase: ++++) was also higher than that of normal endometrium (early proliferative phase: - approximately ++; secretory phase: +++). Higher intensities of ERalpha, ERbeta and 17beta-HSDIand lower intensities of 17beta HSDII were observed in EMI than in the eutopic endometrium of adenomyosis. CONCLUSIONS: The elevation of ERalpha positive cell number, 17beta-HSDI level as well as the insufficient compensation of 17beta-HSDII in eutopic endometrium with adenomyosis and the change in expression pattern of ERalpha, ERbeta, 17beta-HSDI and 17beta-HSDII in EMI lead to the local enhancement of estrogen effect, which would promote cell proliferation. PMID- 15130352 TI - [Effects of progestin on proliferation and differentiation of osteoblasts]. AB - OBJECTIVE: To study the effects of progesterone and levonorgestrel (LNG) on proliferation and differentiation of human normal osteoblasts and human osteosarcoma cell line MG-63. METHODS: Cell proliferation was tested by 3-[4,5 Dimethylthiazolzyl]-2,5-Diphenyl Tetrazolium Bromide (MTT) assay. The expression of c-fos, c-jun and osteocalcin was measured by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) or immunocytochemistry. RESULTS: Compared to the control group, progesterone at concentrations of 10(-10) mol/L, 10(-8) mol/L and 10(-6) mol/L increased the proliferation of human osteoblasts by 9.8%, 23.0% and 32.8% respectively and that of MG-63 cells by 7.8%, 15.6% and 21.8% respectively. LNG at concentrations of 10(-10) mol/L, 10(-8) mol/L and 10( 6) mol/L increased the proliferation of human osteoblasts by 12.5%, 21.9% and 32.8% respectively and that of MG-63 cells by 9.6%, 14.4% and 23.0% respectively. Progesterone and LNG upregulated c-fos and c-jun mRNA and protein in a dose dependent manner. Likewise, progesterone and LNG increased the expression of osteocalcin mRNA in human osteoblasts by 12.2%, 23.7%, 45.5% and 18.6%, 32.1%, 60.3% respectively. The effects of LNG were greater than those of progesterone. Meanwhile, the expression of osteocalcin mRNA in MG-63 cells was not influenced by either progesterone or LNG. CONCLUSION: Progesterone and LNG stimulate proliferation of osteoblasts and increase the expression of c-fos, c-jun and osteocalcin. PMID- 15130353 TI - [Study of neural expression in vaginal mucosa of patients with urinary stress incontinence]. AB - OBJECTIVE: The aim of our study was to investigate the histological changes of vaginal mucosa innervation in stress urinary incontinence (SUI), to determine the alteration that contributed to them. METHODS: Biopsy specimens of anterior vaginal wall were obtained from 56 subjects during operation, which belonged to one of the 3 groups: SUI (15 cases), POP (23 cases) and asymptomatic control (15 cases). Using immunohistochemical method we could semiquantitatively determine the expression of neurofilament (NF) in vaginal mucosa as the distribution of the nerve terminals. RESULTS: There were brown-stained nerve fibers in the lamina propria. According to the expression staging (-, +, ++, +++), the number of patients was 6, 5, 4, 0 cases in SUI group, 17, 5, 1, 0 cases in POP group, and 3, 2, 4, 6 cases in control group, respectively. There was significant difference between SUI group and control group in the expression of NF (P < 0.05) and the expression of nerve fiber had no correlation with the clinical severity and leak point pressure (LPP) in SUI group (P > 0.05). The expression of NF in POP group was also weaker than control. CONCLUSION: The SUI group had a significant less total innervation of vaginal mucosa than asymptomatic controls, which may contribute to the pathogenesis of SUI. PMID- 15130354 TI - [Investigation of urinary incontinence in community dwelling women of Fuzhou city]. AB - OBJECTIVE: The aim was to assess the prevalence of urinary incontinence in Gulou community dwelling women in Fuzhou China. METHODS: One of the communities was selected at random and questionnaires were sent to women living there. After completed the questionnaires were collected and statistics software SPSS 10.0 was used for analysis. RESULTS: We sent 5871 questionnaires and collected 5392 ones. The response rate was 91.8%. The prevalence of urinary incontinence was 18.5%, while it was 51.1% in women who were 65 years or older, and the prevalence of stress urinary incontinence (SUI), urge urinary incontinence (UUI) and mix urinary incontinence (MUI) was 9.1%, 2.4%, and 7.0% respectively. Their structural proportion was 49%:13%:38%. In terms of the incidence of the disease, mild urinary incontinence was prominent. Among the surveyed women, 32.4% did not know where to consult a doctor for help, 45.5%went to urologists, and 22.1% went to gynecologists for help. CONCLUSION: Urinary incontinence is highly prevalent, especially among aged women. However, they seldom consult a doctor about their urinary leakage. PMID- 15130355 TI - [Benefit of palliative surgery for bowel obstruction in recurrent ovarian carcinoma]. AB - OBJECTIVE: Intestinal obstruction is a frequent sequela of recurrent ovarian cancer and difficult to deal with. We analyzed a series of such patients to determine if their outcomes have changed after undergoing palliative surgery. METHOD: We retrospectively reviewed 67 patients undergoing surgery for intestinal obstruction due to recurrent ovarian carcinoma and 75 patients receiving non surgical treatment from 1997 to 2002. RESULTS: During the study period, 67 operations were performed on 67 patients. Among them, surgical procedure was completed in 58 cases. Successful palliation was achieved in 64.2% of cases in which surgical correction was possible. The median survival of the entire cohort was 7.8 months, and 12.6 months for the surgically successfully relieved patients and 3.7 months for those non-surgical patients. The rate of major surgical morbidities was 22.4%. The perioperative mortality rate was 6.0%. Successful palliation was associated with the absence of two prognostic factors: multiple obstructive sites and palpable abdominal and pelvic masses. CONCLUSION: Palliative surgery for bowel obstruction in recurrent ovarian cancer can be worthwhile, and properly selected patients are the key to its success. PMID- 15130368 TI - [Thinking on issues of diagnosis and treatment of endometrial carcinoma]. PMID- 15130369 TI - [Role of three-dimensional transvaginal sonography in evaluation of the depth of myometrial invasion of endometrial carcinoma]. AB - OBJECTIVE: To investigate the value of three-dimensional transvaginal sonography (3-DTVS) in diagnosing depth of myometrial invasion (MI) and analyze factors that may influence 3-DTVS diagnosis. METHODS: Fifty-three patients with endometrial carcinoma proven by histological diagnosis postoperatively in Women's Hospital, Zhejiang University from 2002 to 2003 were included in the study. All patients underwent primary surgery and 2-DTVS and 3-DTVS examinations within 7 days before operation. The diagnosis of the depth of MI was made by multiplanar mode reconstruction and volume measurement based on 3-DTVS. Clinical-pathological parameters were simultaneously recorded. RESULTS: Eighteen patients were deep MI, 31 superficial MI and 4 no myometrial involvement. The sensitivity, specificity, positive and negative predictive values of 3-DTVS and 2-DTVS in detecting superficial MI were 92%, 100%, 100%, 67%; and 44%, 100%, 100%, 21%, respectively. The sensitivity, specificity, positive and negative predictive values of 3-DTVS and 2-DTVS in detecting deep MI were 72%, 86%, 72%, 86% and 75%, 84%, 67%, 89%, respectively. There was significant difference between 3-DTVS and 2-DTVS in detecting superficial MI (chi(2) = 13.2011, P = 0.005), but no significant difference in detecting deep MI (chi(2) = 0.0000, P > 0.05). The median tumor volume of deep and superficial MI was 1.12 cm(3) (Q(25 - 75) = 1.12 - 4.49) and 9.16 cm(3) (Q(25 - 75) = 3.35 - 23.12) respectively. There was a significant difference between the two groups (z = -3.72, P = 0.000). Among all the parameters involved in the study there was no significant factor influencing 3 DTVS diagnosis. CONCLUSIONS: Three-DTVS is superior to 2-DTVS in detecting superficial MI, but not in deep MI. The measurement of tumor volume with 3-DTVS could be used as an objective parameter for detecting deep MI. PMID- 15130371 TI - [Study of different surgeries for clinical stage I endometrial carcinoma]. AB - OBJECTIVE: To compare the outcomes and the risk of post-operative complications in patients with stage I endometrial carcinoma who were treated with different surgeries. METHODS: A total of 211 cases with stage I endometrial cancer treated with surgery in our Cancer Center from Jan 1986 to Dec 1997 were analyzed retrospectively. Sixty-one patients (group 1) underwent simple hysterectomy and salpingo-oophorectomy and 150 patients (group 2) underwent radical hysterectomy. The 5-year survival rates and the risk of post-operative complications were compared between two groups. RESULTS: Five-year survival rates of the group 1 and 2 were 96.0% and 93.5% (P > 0.05), respectively. The recurrence rates of the two groups were 6.6% and 10.7% (P > 0.05), respectively. The overall rates of post operative complications in the two groups were 11.5% and 24.7% (P < 0.05), respectively. CONCLUSION: The patients with stage I endometrial carcinoma who were treated with simple hysterectomy and salpingo-oophorectomy did almost as well as those who underwent radical hysterectomy. PMID- 15130370 TI - [Effect of pelvic lymphadenectomy on prognosis of endometrial carcinoma]. AB - OBJECTIVE: To study the associated factors with pelvic lymph node metastasis of endometrial carcinoma and the effect of pelvic lymphadenectomy on prognosis of the disease. METHODS: Totally 102 patients with endometrial carcinoma who underwent pelvic lymphadenectomy (90 patients) or lymph node biopsy (12 patients) in our hospital from Jan 1981 to Dec 2002 were recruited. The relationship between various clinicopathologic factors and pelvic lymph node metastasis was analyzed. Prognosis of ninety patients with pelvic lymphadenectomy was compared with 90 patients without pelvic lymphadenectomy (control group) in the same period. The 5-year survival was calculated by life table method. RESULTS: The incidence of pelvic lymph node metastasis increased in patients with low grade (46%), deep myometrium invasion (42%), cervical involvement (44%), positive peritoneal cytology (52%), adenexal metastasis (75%) and distant spread (100%). The 5-year survival was lower in patients with lymph node metastasis (37%) than that without lymph node metastasis (89%, P < 0.05). Univariate and COX regression analysis demonstrated that pelvic lymphadenectomy did not improve patients' prognosis. The 5-year survival in patients undergoing lymphadenectomy was 78%, and it was 72% in patients without lymphadenectomy. CONCLUSIONS: The high risk factors for pelvic lymph node metastasis in endometrial carcinoma include low grade differentiation deep myometrium invasion, cervical involvement, positive peritoneal cytology, adenexal metastasis and distant spread. The prognosis is poorer in patients with pelvic lymph node metastasis. Pelvic lymphadenectomy could not improve the prognosis of patients with endometrial carcinoma. PMID- 15130373 TI - [Clinical analysis of endometrial carcinoma patients aged 45 years and younger]. AB - OBJECTIVE: To retrospectively analyze the clinical characteristics and outcomes of endometrial carcinoma patients aged 45 years and younger. METHODS: Fifty-two cases of endometrial carcinoma aged 45 years and younger were treated in Peking Union Medical College Hospital. They were further divided into group A (35 years of age and younger) and group B (older than 35 years). Clinical data of these patients were reviewed and the two groups were compared. RESULTS: Patients aged 45 years and younger accounted for 12.7% of all the endometrial carcinoma cases. About 50% of the patients were nulliparous, infertile or had irregular menstruation and endometrial hyperplasia, 29% were obese, 23% had polycystic ovaries. Eighty-three percent of the patients were stage [International Federation of Gynecology and Obstetrics (FIGO), 1988]. Group A had more polycystic ovaries and atypical endometrial hyperplasia than group B (53% vs 9%, 59% vs 26% respectively, P < 0.05). All group A patients were stage I endometrial carcinoma. In group B, 26% had high risk factors, and compared with group A, FIGO stage was higher (P < 0.05). Operation was the main treatment. Two patients were treated successfully with conservative high dose progestin. Two patients relapsed. CONCLUSIONS: There were high incidences of infertility, irregular menstruation, endometrial hyperplasia, obese and polycystic ovaries in patients aged 45 years and younger, indicating the relationship between endometrial carcinoma and estrogen. Most patients, especially those younger than 35 years, were stage I with few risk factors and good prognosis. Conservation of fertility and ovarian function should be considered in these patients. PMID- 15130374 TI - [Analysis of the risk factors for ovarian metastasis in patients with endometrial carcinoma]. AB - OBJECTIVE: To study the risk factors for ovarian metastasis in patients with endometrial carcinoma. METHOD: The pathological and clinical features and outcomes of endometrial carcinoma patients who were diagnosed and treated in our hospital from Jan 1996 to Dec 2002 were retrospectively reviewed and analyzed. RESULTS: Of the 321 cases reviewed, 15 (4.7%) had ovarian metastasis, among which 60% were recessive metastasis. Factors predictive of ovarian metastasis on logistic forward regression were depth of myometrial invasion, histologic grade and regional lymphatic nodes invasion. CONCLUSION: Depth of myometrial invasion, histologic grade and regional lymphatic nodes invasion are the independent predictive factors for ovarian metastasis of endometrial carcinoma. PMID- 15130375 TI - [Comparative analysis of laparoscopic surgery and laparotomy for early stage endometrial cancer]. AB - OBJECTIVE: To evaluate the feasibility of laparoscopic surgical treatment of early stage endometrial cancer. METHODS: From January 1998 to August 2003, 24 endometrial cancer cases treated by laparoscopy were analyzed as the study group. And 41 endometrial cancer cases treated by laparotomy during the same period were randomly selected as the control group. The two groups were compared in terms of the clinic data of perioperative periods. RESULTS: The clinicopathological characteristics before operation between both groups were similar. The mean operating time in the laparoscopy group (97 minutes) was significantly shorter (P < 0.001) than that in the laparotomy group (134 minutes). The blood loss during the operation in the laparoscopy group (163 ml) was fewer than that in the laparotomy group (259 ml). The numbers of the lymph nodes resected between the two groups were similar. The rate of complications in the laparoscopy group was lower than that in the laparotomy group. The laparoscopy group had shorter hospitalization (6.3 days) than that of the laparotomy group (9.6 days, P < 0.01). The pathological type and International Federation of Gynecology and Obstetrics (FIGO) stage between the two groups were similar. One case had recurrence and 1 case died in the control group but there was no such case in the laparoscopy group. The survival rate of the study group was 100% and that of the control group was 97% (P > 0.05). CONCLUSION: Laparoscopic surgery is feasible and safe in treatment of early stage endometrial cancer, and the clinic value should be confirmed by multicenter randomized clinic trial. PMID- 15130376 TI - [Study of teratogenicity of hyperglycemia on neural tube defects and antagonistic effect of taurine]. AB - OBJECTIVE: To explore the molecular mechanism of neural tube defects (NTDs) caused by hyperglycemia and thiadiazole and the antagonistic effect of taurine. METHODS: The pregnant mice were divided into hyperglycemia groups, thiadiazole group, taurine groups and control groups. The mRNA and the protein of Pax3 or Cx43 gene were detected respectively by reverse transcription-polymerase chain reaction assay and immunohistochemical method. RESULTS: As compared with mice treated by thiadiazole-stomach-perfusing, NTDs were significantly increased from mice treated with glucose-injection when blood glucose levels were >or= 13.4 mmol/L. Elevated glucose and thiadiazole could cause changes in Pax3 and Cx43 expression. Hyperglycemia had stronger developmental toxicity on mice embryos. Expression of Pax3 (mRNA 0.97 +/- 0.20, protein 0.11 +/- 0.02) in hyperglycemia group was significantly decreased, while expression of Cx43 (mRNA 7.05 +/- 1.63, protein 0.94 +/- 0.05) was significantly increased, and the relationship of dose effect was demonstrated. In the thiadiazole group, the expression of Cx43 (mRNA 6.96 +/- 0.73, protein 0.92 +/- 0.12) was significantly stronger than control groups, but there were no significant differences in expression of Pax3 between thiadiazole and its control groups. Both of their teratogenicity could be antagonized by taurine. CONCLUSIONS: This study suggests that congenital malformation associated with diabetic pregnancy is caused by disruption of regulatory genes, Pax3 and Cx43 expression in embryo in response to elevated glucose. Thiadiazole can only disturb the regulation of Cx43 gene causing NTDs. Taurine can correct the disruption caused by the two teratogens. PMID- 15130377 TI - [Effect of rosiglitazone on ovulation induction in women with polycystic ovary syndrome]. AB - OBJECTIVE: To investigate the effect of rosiglitazone on ovulation induction in women with polycystic ovary syndrome (PCOS). METHODS: Ninety-six PCOS women with insulin resistance (IR) were randomly divided into three groups. Group A (28 cases) received clomiphene citrate (CC) alone. Group B (32 cases) received rosiglitazone alone. And group C (36 cases) received CC and rosiglitazone in combination. All were treated for three menstrual cycles. Homeostasis model assessment insulin resistance (HOMA IR) and ovulation before and after treatment were compared among the 3 groups. RESULTS: After the treatment, HOMA IR in group B and C decreased from 1.2 +/- 0.6 to 0.6 +/- 0.2 and from 1.1 +/- 0.5 to 0.6 +/- 0.4, respectively (P < 0.05). Success rate of ovulation after treatment in group C (80%) was higher than that in group A (59%) and group B (35%, P < 0.05). CONCLUSION: Rosiglitazone therapy improves insulin sensitivity and ovulation induction. PMID- 15130378 TI - [Tyrosine phosphorylation and protein expression of insulin receptor substrate-1 in the patients with polycystic ovary syndrome]. AB - OBJECTIVE: To investigate the tyrosine phosphorylation and protein expression of insulin receptor substrate-1 in adipose tissue from patients with polycystic ovary syndrome, and explore molecular mechanisms of insulin resistance of PCOS. METHODS: Samples from patients with PCOS with insulin resistance (group A, n = 19), PCOS without insulin resistance (group B, n = 10) and controls group (n = 15) were collected. Serum luteinizing hormone (LH), follicle stimulating hormone (FSH) and testosterone (T) were measured by chemiluminescence assay. Fasting insulin (FIN) was measured by radioimmunoassay. Fasting plasma glucose (FPG) was measured by oxidase assay. Insulin resistance index was calculated using homeostasis model assessment (HOMA) to analyze the relationship between these markers and insulin resistance. The amount of insulin receptor substrate-1 in adipose tissue was assessed by western blot. The tyrosine phosphorylation of IRS 1 was measured by immunoprecipitation. RESULTS: (1) The levels of serum LH (15.8 +/- 2.8) U/L, LH/FSH 2.8 +/- 0.6, T (4.3 +/- 0.9) nmol/L, FIN (25.2 +/- 3.8) mU/L and HOMA IR (1.56 +/- 0.25) in group A were significantly higher than those of group B (13.9 +/- 1.9) U/L, 2.3 +/- 0.4, (3.6 +/- 0.4) nmol/L, (13.4 +/- 3.8) mU/L, 0.87 +/- 0.28 and control group (7.3 +/- 2.1) U/L, 1.3 +/- 0.3, (0.9 +/- 0.2) nmol/L, (9.5 +/- 2.6) mU/L, 0.50 +/- 0.30 (all P < 0.05); (2) The protein expression and tyrosine phosphorylation of IRS-1 in group A (690 +/- 19 and 528 +/- 72 respectively) were significantly lower than those in group B (892 +/- 31, 801 +/- 64) and control group (988 +/- 29, 1139 +/- 124) (P < 0.05, and P < 0.01 respectively). (3) Insulin resistance index in group A and group B were negatively related with protein expression and tyrosine phosphorylation (r = 0.52, P < 0.05; r = -0.61, P < 0.01 and r = -0.60, P < 0.05; r = -0.63, P < 0.05). CONCLUSIONS: The signal transduction malfunction because of protein expression and tyrosine phosphorylation changes of IRS-1 in adipose tissue from polycystic ovary syndrome patients may be one of the mechanisms leading to insulin resistance. PMID- 15130379 TI - [Clinical analysis of ultrasound-guided embryo transfer after in-vitro fertilization]. AB - OBJECTIVE: To investigate whether ultrasound (US) guided embryo transfer could improve pregnancy rates. METHODS: A prospective randomized trial was conducted to compare embryo transfer under abdominal US guidance (n = 178) with traditional embryo transfer (n = 152). RESULTS: The clinical pregnancy rate was 37.1% (66/178) in the US group compared with 25.0% (38/152) in the traditional group (P < 0.05). CONCLUSIONS: US guided embryo transfer after in vitro fertilization increased pregnancy and implantation rates. It is suggested that embryo transfer should be performed under ultrasound guidance. PMID- 15130397 TI - [Brain death: a problem has to be faced]. PMID- 15130398 TI - [How to diagnose and treat the Helicobacter pylori infection]. PMID- 15130399 TI - [Legislation of the national brain death act is imminent]. PMID- 15130401 TI - [Imminent issues on clinical diagnostic criteria for brain death]. PMID- 15130403 TI - [Internist physicians are needed in general and specialized hospitals]. PMID- 15130404 TI - [Studies on evaluation of brain death]. AB - OBJECTIVE: To investigate the methods of evaluating brain death accurately and objectively. METHODS: Various kinds of clinical examination [including conscious state, Glasgow coma score (GCS), brain stem reflection, cranial nerve dominated actions, spinal reflection, spinal nerve dominated automatic action, respiratory and apnea testing] and laboratory methods [such as electroencephalography (EEG), brainstem auditory evoked potentral (BAEP), brainstem auditory sensery evoked potentral (SLSEP) and transeranial Doppler (TCD)] were used to evaluate 11 cases with brain death. RESULTS: Eleven cases presenting with deep coma were conssedered to have brain death. All of them had GCS score of 3 and disappearance of brain stem reflection and cranial nerve dominated automatic action. Six of them patients showed anisocoria. Nine of them pharyngeal reflex could not be detected because of tracheal cannulization, but this did not influence the evaluation of brain death. None of them showed "flat" EEG and disappearience of BAEP dominant wave. None of the SLSEP show any wave behind N13. TCD showed blood stream characteristic of brain death except in one case blood stream characteristic of increased intracraniai pressure. Apnea test and atropine test provided the final proof brain death. The former had more influence on heart rate, blood pressure and oxygen saturation while the latter were more safe and reliable. CONCLUSION: Combined and continuous observation of clinical and laboratory indices can enhance the accuracy of the evaluation of brain death. PMID- 15130406 TI - [Biodiv Ysio stent for symptomatic stenosis of middle cerebral artery]. AB - OBJECTIVE: To assess the safety and feasibility of Biodiv Ysio stent for patients with symptomatic M1 stenosis of middle cerebral artery (MCA). METHODS: Forty three patients with forty-five M1 stenoses, ranged from 50% to 99% in diameter reduction and refractory to medical therapy, were enrolled in this study between March 2002 and November 2003. The lesions were situated at M1 trunk (n = 19), M1 origin (n = 12) and M1 bifurcation (n = 14), respectively. RESULTS: The technical successful rate was 97.8% (44/45) for all the lesions. The rate of complicating subarachnoid hemorrhage was 7.0% (3/43 patients) and the rate of death was 2.3%. During a follow-up of 7.0 months (median), there was no recurrence of transient ischemic attack or stroke in 41 available patients. Six-month angiographic follow up was obtained in 6 patients (7 vessels), demonstrating good patency in 6 stenting vessels and restenosis in one vessel. CONCLUSIONS: Angioplasty associated with Biodiv Ysio stent appears to be a safe and feasible for the patients with symptomatic M1 stenosis of MCA, under strict control of periperformeral project. Further study is, however needed. PMID- 15130407 TI - [The efficiency and safety of anticoagulation therapy in atrial fibrillation in Chinese]. AB - OBJECTIVE: To determine the strength of oral anticoagulation therapy in atrial fibrillation that provides the best balance between the prevention of thromboembolism and the occurrence of bleeding complications. METHODS: We studied 435 patients with atrial fibrillation who were hospitalized from 2000 to 2002 and given warfarin for prevention of thromboembolism. INR-specified rates for both ischemic and major hemorrhagic events were analyzed and the optimal levels of anticoagulation in atrial fibrillation patients determined. RESULTS: The average dose of warfarin was (2.77 +/- 0.83) mg and the median duration of anticoagulation is 7 months (from 1 month to 3 years). In total, there were 31 confirmed bleeding events, with major hemorrhage occurring in 5 patients. Age of the patients in the hemorrhage group is not significantly higher than that in control group (65.09 +/- 9.99 vs 62.01 +/- 12.19, P = 0.259). Chronic heart failure or hypertension increased the risk of bleeding during warfarin therapy. Multivariate analysis showed that INR >or= 3.0 is an independent risk factor for hemorrhage (OR = 3.7435, 95% CI 1.2819 - 8.9838). The risk of stroke or thromboembolism rose steeply with INR below 1.5. CONCLUSIONS: To achieve optimal levels of anticoagulation with the lowest risk in patients with atrial fibrillation, values of INR below 1.5 and above 3.0 should be avoided. PMID- 15130408 TI - [Effects of hypercholesterolemia on the number and activity of endothelial progenitor cells in peripheral blood]. AB - OBJECTIVE: To investigate whether hypercholesterolemia has influences on the number and activity of endothelial progenitor cells (EPCs). METHODS: Mononuclear cells were isolated from patients with hypercholesterolemia (n = 20) and age matched control subjects (n = 20). EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. EPCs proliferation and migration were assayed by MTT assay and modified Boyden chamber assay, respectively. EPCs adhesion assay was performed by replating cells on fibronectin coated dishes, then counting the adherent cells. RESULTS: The number of EPCs was significantly reduced in patients with hypercholesterolemia as compared with that in control subjects [(41.8 +/- 8.7 vs 64.5 +/- 16.6) EPCs/x 200 fields; P < 0.05] and it was inversely correlated with total cholesterol levels (r = -0.659, P < 0.001) and LDL cholesterol levels (r = -0.611, P < 0.001). In addition, EPCs proliferative, migratory and adhesive capacity were also impaired. CONCLUSION: It is suggested that a novel pathophysiological mechanism of hypercholesterolemia may be defined i.e. reduction of EPCs with decreased functional activity. PMID- 15130409 TI - [Expression pattern of tumor-associated antigen MG7-Ag in gastric cancer: difference between Tibetans and Hans]. AB - OBJECTIVE: To investigate the difference of the expression pattern of tumor associated antigen MG(7)-Ag in gastric carcinoma between Hans and Tibetans. METHODS: Avidin-biotin peroxidase complex immunohistochemical methods were adapted to examine the expression pattern of MG(7)-Ag in 200 patients with gastric carcinoma, including 100 Tibetan patients and 100 Hans patients, 10 patients with chronic artrophic gastritis, 18 with gastric mucosal dysphasia and 10 subjects with normal gastric mucosa served as control. RESULTS: The positive rate for MG(7)-Ag was 92.4% and 88.7% in Tibetan and Han gastric carcinoma patients, 55.6% and 38.9% in Tibetan and Han gastric mucosal dysplasia patients, 20.0% and 10.0% in Tibetan and Han chronic atrophic gastritis patients, respectively. There was no positive expression in normal gastric mucosa of both Hans and Tibetans. >From normal gastric mucosa, chronic artrophic gastritis and gastric mucosal dysplasia to gastric carcinoma, expression of MG(7)-Ag showed an ascending tendency in the same group of subjects or patients (P < 0.01). However between Hans and Tibetans there was no significantly different expression of MG(7)-Ag. In gastric carcinoma, the expression pattern of MG(7)-Ag was not related to tumor cell differentiation and metastasis of lymph nodes in Hans and Tibetans (P > 0.05), but in the same group the expression of MG(7)-Ag was related to metastasis of lymph nodes. CONCLUSIONS: The expression pattern of MG(7)-Ag displayed no significant difference between Hans and Tibetans, The tumor associated antigen MG(7)-Ag in gastric carcinoma can be used in Tibetans as a reliable marker to predict early gastric cancer. PMID- 15130410 TI - [The role of duodenogastroesophageal reflux in gastroesophageal reflux disease]. AB - OBJECTIVE: To investigate the role of duodenogastroesophageal reflux (DGER) in the pathogenesis of gastroesophageal reflux disease (GERD) and its value for the diagnosis of non-erosive reflux disease (NERD). METHODS: 95 cases of GERD were divided into two groups according to endoscopic findings: reflux esophagitis (RE) (n = 51) and NERD (n = 44). Simultaneous 24-hour esophageal pH and bilirubin monitoring were performed. RESULTS: The index values of DGER in RE group such as percent of time with Abs > 0.14, total reflux time and time with reflux > 5 min were 19.05 +/- 23.44, 30.56 +/- 34.04 and 5.90 +/- 6.37 respectively, they were significantly higher than those of NERD, 7.26 +/- 11.08, 15.68 +/- 20.92 and 2.59 +/- 3.57, respectively (P < 0.05), but no significant difference was found in acid reflux. The occurrence of DGER was more common in more severe esophagitis. Isolated DGER were found in 18.2% of NERD, and combined bilirubin monitoring could elevate the positive diagnosis rate of NERD from 65.9% to 84.1%. CONCLUSIONS: DGER could be present isolatedly and played an obvious role in the genesis of RE and GERD symptoms. Simultaneous 24-hour esophageal pH and bilirubin monitoring were helpful for the diagnosis of NERD. PMID- 15130411 TI - [The mechanism and implication of costimulatory molecules in dendritic cells in the development of asthma]. AB - OBJECTIVE: To exploring the role of costimulatory molecules in the development of asthma and its mechanisms. METHODS: Murine asthma model was established with ovalbumin (OVA) sensitization and challenge, and the model was confirmed by histological analysis of lung tissues, cell numbers and differentiations of bronchoalveolar lavage, serum OVA-specific IgE level and interleukin (IL)-4 and IL-5 production by splenic T cells. The purity of spleen-derived dendritic cells was assayed with fluorescein-actived cell sorter (FACS) by analyzing CD(11c) molecule. FACS was also used to measure the expression of CD(80) and CD(86) on spleen-derived dendritic cell (DC) from OVA-sensitized and challenged mice. Finally, the production of IL-4 and IL-5 in naive T cells after stimulation with spleen-derived DC from OVA-sensitized and challenged mice were determined with ELISA. RESULTS: Histological analysis of lung tissues, components of broncho alveolar fluid, the level of serum OVA-specific IgE, and the production of IL-4 and IL-5 were all consistent with the characteristic of a murine asthma model. The expression of CD(80) on spleen-derived DC from OVA-sensitized and challenged mice was increased significantly compared with that from PBS-treated mice, while there was no difference in CD(86) expression. On the other hand, DC from OVA sensitized and challenged mice stimulated the production of IL-4 and IL-5 by naive T cells. CONCLUSION: Our results suggest that DC, via upregulation of CD(80), might play a pivotal role in the maintenance and amplification of allergic immune response, namely the Th2 immune response. PMID- 15130412 TI - [Bone mineral density and leptin receptor polymorphism Gln223Arg in Han women in Beijing]. AB - OBJECTIVE: To investigate the association between bone mineral density (BMD) and leptin receptor (LEPR) polymorphism (Gln223 Arg) in young women and postmenopausl osteoporotic women. METHODS: BMD values were determined by dual energy X-ray absorptiometry. The Gln223 Arg genotypes of LEPR were analyzed by using polymerase chain reaction-restriction fragment length polymorphism method. RESULTS: Hardy-Weinberg equilibrium was evident for LEPR polymorphism. The subjects carrying the GG genotype of LEPR had significantly higher BMD at lumbar spine as compared with the subjects with GA and AA genotype in young women [(1.213 +/- 0.127) g/cm(2) vs (1.154 +/- 0.124) g/cm(2), P < 0.05]. There were no significant differences in BMD at the proximal femur among GG, GA and AA genotype in young women. No significant differences in BMD at all sites were observed among GG, GA and AA genotype in postmenopausl osteoporotic women. Correlation was found between BMD and leptin receptor polymorphism (Gln223 Arg) at lumbar spine (r = -0.151, P < 0.05). CONCLUSION: Leptin receptor polymorphism (Gln223 Arg) has association with peak bone mass in young women, which may be used as genetic marker in predicting the risk of developing osteoporosis in Chinese women of Han nationality. PMID- 15130413 TI - [Effects of rosiglitazone and metformin on insulin resistance in high-fat diet rats]. AB - OBJECTIVE: To observe the effects of rosiglitazone and metformin in rats on insulin resistance induced by high-fat diet. METHODS: Normal 8-week old male SD rats were divided into four groups. They were normal chow group (NC, n = 11), high-fat diet (HF, n = 11), metformin-treated (HF + Met, n = 11) and rosiglitazone-treated group (HF + Ros, n = 11). Rosiglitazone 3 mg x kg(-1) x d( 1) and metformin 300 mg x kg(-1) x d(-1) were given orally to HF + Ros and HF + Met group, respectively. After feeding for 8 weeks, serum insulin, adiponectin, glucose (BG), triglyceride (TG) and free fatty acid (FFA) were measured in all the rats. Insulin sensitivity was measured with glucose infusion rate (GIR) and determined by using euglycemic-hyperinsulinemic clamp method. RESULTS: High-fat diet induced obesity in SD rats after feeding for 8 weeks. High-fat diet decreased adiponectin level by 43.7% (P < 0.01) and GIR by 51.3% (P < 0.01) as compared with the NC group. Metformin decreased body weight by 8.4% (P < 0.01) and TG level by 40.5% (P < 0.01). Metformin significantly increased GIR by 58.9% (P < 0.01) when compared with the HF group. Rosiglitazone caused an apparent reduction of FFA (-25.3%, P < 0.05) and TG level (-54.0%, P < 0.01). At the same time, rosiglitazone increased adiponectin by 60% (P < 0.01), and improved insulin sensitivity by 149.6% (P < 0.01) as compared with the HF group. CONCLUSIONS: (1) High-fat diet induces insulin resistance in SD rats; this was associated with an increase in visceral fat and a decrease in the level of adiponectin; (2) Metformin treatment improved insulin sensitivity accompanied by a decrease in body weight and TG level; (3) Rosiglitazone treatment ameliorates IR in a greater extent and is accompanied by a reduction of FFA, TG and an increase of adiponectin levels. PMID- 15130414 TI - [The clinical application of 18F-fluorodeoxyglucose coincidence imaging in lymphoma]. AB - OBJECTIVE: To investigate clinical value of (18)F-fluorodeoxyglucose ((18)F-FDG) coincidence imaging (SPECT/PET) in diagnosis and treatment of lymphoma. METHODS: (18)F-FDG metabolic imaging was performed in eighteen patients NHL before and after treatment using a Vertex(Plus) EPIC MCD/AC from ADAC Company. The findings of (18)F-FDG metabolic images were analyzed in qualitative and semi-quantitative ways. (18)F-FDG images were compared with synchronous routine images such as ultrasonography and CT. RESULTS: Thirty studies were performed in 18 patients with NHL in this study. There were 21 true positives, 6 true negatives, 2 false positives and 1 false negative. The accuracy, sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) were 90.0%, 95.5%, 75.0%, 91.3%, and 85.7%, respectively. (18)F-FDG imaging led to a change of 16.7% in clinical staging, and 46.7% in management. The PPV and NPV of recurrence detected by (18)F-FDG SPECT/PET were 100% and 80.0%, while these of CT were 50.0% and 25.0% after therapy. Four of five patients with negative entered clinical complete remission (CR), whereas all the 11 patients with abnormal (18)F-FDG uptake relapsed or reprogressed. Progression-free survival (PFS) with negative (18)F-FDG was 16 - 47 months (median: 28.7 months) and PFS with positive was 3 - 46 months (median: 8.3 months). CONCLUSION: SPECT/PET (18)F-FDG metabolic imaging plays an important role in clinical diagnosis and treatment of NHL. PMID- 15130416 TI - [The expression and significance of major histocompatibility complex II transactivator gene in five human malignant hematological cell lines]. AB - OBJECTIVE: To investigate the role of major histocompatibility complex (MHC) II transactivator in constitutive or interferon (IFN)-gamma inducible expression of MHC molecules in human malignant hematological cell lines. METHODS: The expression of MHC molecules and MHC class II transactivator (CIITA) protein were detected with Western blot, immunohistochemistry and flow cytometry. The expression of CIITA gene was determined with RT-PCR. T cell expressing interleukin (IL)-2 mRNA was induced by Jurkat with mixed lymphocyte reaction. RESULTS: It was found that the MHCII-positive tumor cells expressed the CIITA quite well, and IFN-gamma could induce the expression of MHCI + II only if the tumor cells were able to express CIITA in an IFN-gamma induced manner (20%, 42% for Jurkat cells respectively), while T cell expressed more IL-2 mRNA (51 times) in the case of Jurkat after induction with IFN-gamma. CONCLUSIONS: It is suggested that there is a correlation between the inability of the tumor cells in response to IFN-gamma for MHC expression and the deficiency in the inducible expression of CIITA. This might be due to immune sneaking through the tumor cells to some extent. PMID- 15130417 TI - [Hyperhomocysteinemia, oxidative stress and microinflammation in chronic renal failure: their roles in atherogene]. AB - OBJECTIVE: To investigate the relationship between hyperhomocysteinemia, oxidative stress and microinflammation in patients with chronic renal failure (CRF), and its role in the pathogenesis of atherosclerosis seen in these patients. METHODS: One hundred and fourteen CRF patients and 37 healthy volunteers were involved in the study. The levels of plasma total homocysteine (tHcy) were determined by fluorescence polarization immunoassay. Plasma malondialdehyde (MDA) levels were analyzed by thiobarbituric acid reaction. The activity of glutathione peroxidase (GSHPx) in plasma was measured by spectrophotometry. Serum C-reactive protein (CRP), interleukin (IL)-6 and tumor necrosis factor (TNF)alpha were determined by ELISA. Intima-medial thickness (IMT) of extracranial common carotid artery and the presence of atherosclerotic plaques was determined by using noninvasive high-resolution B-mode ultrasonography. RESULTS: Levels of tHcy, MDA, CRP and TNFalpha in the non dialysis CRF patients were significantly higher than those in the healthy controls and increased with the progression of renal insufficiency, while plasma activity of GSHPx decreased progressively with decline of renal function. Compared with the pre-dialysis patients, hemodialysis patients exhibited higher levels of tHcy, MDA, CRP, IL-6 and TNFalpha, and lower levels of GSHPx activity. There were no significant difference in the levels of tHcy, MDA, CRP, IL-6, TNFalpha, and GSHPx activity between patients dialyzed with hemophan, polysulfone and cellulose triacetate membrane. Prevalence of atherosclerotic plaque and increased IMT in carotid artery were significantly higher in CRF patients compared with the age-matched controls and associated with the levels of tHcy, MDA and CRP. Multivariate stepwise regressive analysis showed that increased IMT in CRF patients was closely related to the levels of tHcy, MDA, CRP, blood glucose, and very low density lipoproteins (R(2) = 0.572, P < 0.001). The factors associated with plasma MDA (representing lipid peroxidation level) were GSHPx activity (standard regressive coefficient, beta = -0.651, P < 0.01), tHcy level (beta = 0.229, P < 0.01), and serum creatinine (beta = 0.163, P < 0.05). The factors associated with serum CRP (representing microinflammation state) were IL 6 (beta = 0.532, P < 0.01), TNFalpha (beta = 0.212, P < 0.01), MDA (beta = 0.209, P < 0.05), and plasma albumin (beta = -0.297, P < 0.05). CONCLUSION: Hyperhomocysteinemia in CRF patients may accelerate oxidative stress, and lipid peroxidation may be involved in the occurrence of microinflammation state. The complex interaction between hyperhomocysteinemia, oxidative stress and microinflammation may result in accelerated atherosclerosis seen in CRF. PMID- 15130432 TI - [Study on lifting-related musculoskeletal disorders among workers in metal processing]. AB - OBJECTIVE: To investigate lifting-related musculoskeletal disorders in metal processing, to analyze the risk factors, and to study the validity and feasibility of using NIOSH lifting equation in China. METHODS: The questionnaires of semi-structured interview, the Ovako Working Posture Analysing System (OWAS) postural analysis and variables of the NIOSH equation were applied to the study. The study population consisted of 69 workers mainly involved in manual materials handling (MMH), categorized as Job A; and 51 machinery workers, served as controls, that were less MMH task involved, as Job B. RESULTS: The prevalence of low back pain (LBP), which was defined at least one episode lasting for more than 24 hours in the past 12 months, were 63.8% and 37.3% for Job A and Job B, respectively. However, the prevalence of LBP lasting for more than a week due to lifting were 26.09% and 5.88% for Job A and B, respectively. The proportion of awkward back postures were found higher in Job A than that of Job B (66% vs 63%, P < 0.05). The NIOSH Lifting Index (LI) was estimated to be 2.4 for Job A, and 0 < LI < 1 for Job B. The analysis of multiple regressions revealed that the repetitiveness of lifting and length of service had greatly attributed to the occurrence of LBP. The "composite load" (object weight x activity repetitiveness) had a significant adverse effect on lower back meaning that the objective weight remains an ingredient part of the risk. CONCLUSION: The occurrence of LBP is not only related to the force load, but the repetitiveness of lifting and awkward postures. The method of OWAS observation and US-NIOSH equation are important tools in assessing characteristics and risk factors of LBP for MMH tasks. Further study aimed at developing an integral scheme for the assessment system is needed. PMID- 15130433 TI - [Relationship of occupational injuries with social and economic factors]. AB - OBJECTIVE: To explore the relationship of occupational injuries with social and economic factors in chemical industry during 2000.01 - 2001.12. METHOD: 1:2 paired case-control study, univariable logistic regression analysis, principal component analysis, and multiple logistic regression analysis were used in this study. RESULTS: Univariable analysis showed that occupational injuries had significant relationship with age, sex, education, employment pattern, technology, workplace, work changing, wage, family income, enterprise scale, enterprise proprietorship, projective device, operation rules, and training rules of work safety. The extracted four principal components (PC(1), PC(2), PC(3) and PC(4), ranked by contribution) gave good expressions to the initial 11 variables. The cumulative proportion of the four principal components reached 77.36%. PC(1) was the indicative factor of occupational injuries, which represented 46.69% information of initial variables. PC(2) was the kinetic factor of occupational injuries. PC(3) was the stable factor of occupational injuries. PC(4) was the sex factor of occupational injuries. The results of multiple conditional logistic regression analysis showed that occupational injuries had statistically significant relationship with PC(1) and PC(2). Among the initial variables, sex, employment pattern, income, scale of enterprise, and property of enterprise were more prominent. CONCLUSION: Occupational injuries are related with multiple social and economic factors, which often interact on each other. The prevention and control of occupational injuries should require a comprehensive approach, including training and education of work safety, improving workers' consciousness of self-protection, and enhancing proprietors' consciousness of work safety. PMID- 15130434 TI - [Inhibitory effects of basic fibroblast growth factor on fibrosis of strain injured skeletal muscles in rats]. AB - OBJECTIVE: To investigate the effects of exogenous basic fibroblast growth factor (bFGF) on the healing of strain injured skeletal muscles in rats. METHODS: Eighteen male SD rats were randomly divided into three groups: no strain control group (C(on)), muscle strained and bFGF treated group (S(b)), or normal saline treated group (S(0)). The gastrocnemius of rats in the S(b) and S(0) groups was strained and the animals were treated with bFGF (200 AU/d) or normal saline for six days. Vimentin expression, an indicator of muscle fibrosis in injured muscles (expressed as integral optical density, IOD), was measured by immunohistochemistry. RESULTS: The IOD of vimentin in the S(0) group [(24.29 +/- 7.91) x 10(3)] was higher than that in the C(on) group [(5.75 +/- 3.87) x 10(3)] (P < 0.01). The IOD of vimentin in bFGF treated group [(15.78 +/- 7.72) x 10(3)] was lower than that in the normal saline treated group (P < 0.01). CONCLUSION: The exogenous bFGF may facilitate the repair of muscle structure and function by reducing vimentin expression and fibrosis in strain injured muscles. PMID- 15130435 TI - [Detecting DNA repair capacity of human lymphocytes exposed to ultraviolet C with comet assay]. AB - OBJECTIVE: To assess DNA repair capacity of human lymphocytes with comet assay. METHODS: Fresh lymphocytes form twelve 26-year old donors (6 males, 6 females) were exposed to ultraviolet C (UVC, 254 nm) at the dose rate of 1.5 J/m(2). The lymphocytes of each donor were divided into three parts: UVC group, UVC + aphidicolin (APC) group, UVC + novobiocin (NOV) group. DNA single strand breaks were detected with comet assay in UVC-irradiated cells and unirradiated cells incubated for 30, 60, 90, 120, 180 and 240 min. DNA repair rate (DRR) was calculated and served as an indicator of DNA repair capacity. RESULTS: The maximum average comet tail length (MTL) in three groups appeared 90 min after UVC exposure. The DRR range of UVC group was 81.84% (62.84% - 98.71%); There was no significant difference in DRR between males and females (P > 0.05). However, the average DRRs of UVC + NOV group and UVC + APC group (52.98% and 39.57% respectively) were significantly lower than that of UVC group (P < 0.01). CONCLUSION: Comet assay is a rapid and simple screening test to assess DNA repair capacity. DRR, as an indicator, may express the individual DNA repair capacity. PMID- 15130436 TI - [Role of heat shock protein 70 expression in DNA damage induced by benzo(a)pyrene]. AB - OBJECTIVE: To explore heat shock protein 70 (HSP70) expression of A549 cells and its role in DNA damage caused by benzo(a)pyrene (BaP). METHODS: Human adenocarcinoma A549 cells were cultured in vitro, exposed by different concentrations of BaP (0, 1.25, 2.50, 5.00, 10.00 micro mol/L) for 6 hours, or 10 micro mol/L of BaP for different time (0, 4, 8, 12, 16, 24 and 48 h). Then HSP70 expression and DNA damage were detected using Western-blot and single cell gel electrophoresis (SCGE) assay respectively, and the relationship between HSP70 expression and DNA damage was further analyzed. RESULTS: The integral optical densities of HSP70 in A549 cells treated with 1.25, 2.50, 5.00 and 10.00 micro mol/L BaP for 6 h (49.63 +/- 1.30, 45.72 +/- 1.03, 40.53 +/- 0.95, 37.50 +/- 1.20 respectively) were lower than that of the control cells (59.43 +/- 1.17) (P < 0.05). When A549 cells were exposed to 10 micro mol/L BaP for 4, 8, 12, 16 h, the integral optical densities of HSP70 were 33.33 +/- 0.80, 29.23 +/- 0.91, 12.51 +/ 0.96, 9.50 +/- 1.25 respectively, and there was an increasing tendency of the expression of HSP70 for 24 - 48 h (20.06 +/- 1.38, 24.51 +/- 1.39), however, all were different from that in control group (56.59 +/- 0.85) (P < 0.05). DNA damage scores in 10(6) A549 cells treated with 2.50, 5.00 and 10.00 micro mol/L BaP for 6 h (23,718 +/- 2,938, 30,128 +/- 2,937, 44,231 +/- 3,846) were significantly higher than that of the control cell (9,615 +/- 1,923) (P < 0.05). When A549 cells were exposed to 10 micro mol/L BaP for 4, 8, 12, 16, 24, 48 h, DNA damage scores (16,667 +/- 4,003, 38,461 +/- 1,924, 5,615 +/- 3,847, 76,282 +/- 2,937, 7,513 +/- 1,110 and 58,975 +/- 9,487) were also higher than that of control group (P < 0.05). There was a negative correlation between DNA damage and the expression of HSP70 when A549 cells were exposed to different concentrations of BaP. CONCLUSION: HSP70 might enhance intracellular defenses against DNA damage induced by BaP. PMID- 15130437 TI - [Effects of mitogen activated protein kinase signal transduction pathways on heat shock protein 70 gene expression in endothelial cells exposed to benzo(a)pryene]. AB - OBJECTIVE: To investigate the effects of mitogen activated protein kinase (MAPK) signal transduction pathways on heat shock protein 70 (HSP70) gene expression in endothelial cells exposed to benzo(a)pryene (BaP). METHODS: Porcine aortic endothelial cells were pre-treated or by PD98059 (10 micro mol/L) or SB203580 (20 micro mol/L) for 1 hour, then treated with different concentrations of BaP (0, 0.1, 0.5, 1.0, 5.0 and 10.0 micro mol/L) for 24 hours respectively;Expression levels of three phosphorylated MAPKs [extracellular signal regulated protein kinase (ERK), c-Jun amino-terminal kinase (JNK), and p38] and HSP70 were determined by Western-blot. RESULTS: The three phosphorylated MAPKs expressional levels especially p-ERK1 had different extents of changes with dose-response relationship under BaP exposure. BaP inhibited the expression of HSP70, which significantly decreased in medium and high dose group (>or= 1.0 micro mol/L) but did not decrease in control group (P < 0.05). Although the inhibitor of ERK (PD98059) could partly weaken the inhibited effects of BaP on HSP70 expression, HSP70 expression levels of endothelial cells pre-treated with PD98059 were still significantly lower than that of control cells (P < 0.05). CONCLUSION: ERK1 pathway might play some roles in HSP70 gene expression in endothelial cells exposed to BaP, and other unknown signal pathways might also have some effects on this process. PMID- 15130438 TI - [Effect of microwave irradiation on neurocyte mitochondrial ultrastructure and mtTFA mRNA expression in rats cerebral cortex and hippocampus]. AB - OBJECTIVE: To explore molecular controlling mechanism of mitochondrial injury induced by different density of microwave irradiation. METHODS: Rats were exposed to microwave irradiation for 1 hour at average power density of 3 mW/cm(2) or 30 mW/cm(2). After microwave irradiation, the changes of pathological ultrastructure of rat cerebral cortex and hippocampus were observed by electron microscope, and mitochondrial transcription factor A (mtTFA) mRNA expression level were determined by RT-PCR. RESULTS: After 3 mW/cm(2) microwave irradiation for 0, 3, 24 h, mitochondrial ultrastructure and mtTFA mRNA expression level didn't significantly change in rat cerebral cortex and hippocampus. After 30 mW/cm(2) microwave irradiation for 0, 3, 24 h, mitochondrial ultrastructure obviously changed, mtTFA mRNA expression in rat hippocampus significantly increased by 67.00%, 80.00%, 30.00% respectively, and in rat cerebral cortex by 133.00%, 86.00%, 233.00% respectively. There were significant differences between the corresponding groups of hippocampus and cerebral cortex (P < 0.01). CONCLUSION: No obvious change in mitochondria was found after 3 mW/cm(2) microwave irradiation, but it was found after 30 mW/cm(2) microwave irradiation. Mitochondria injury in cerebral cortex was more severe than that in hippocampus. mtTFA mRNA may have certain regulation in mitochondrial energy metabolism. PMID- 15130439 TI - [Effects of microwave radiation on thymocytes in mice at different power densities]. AB - OBJECTIVE: To investigate the effects of microwave radiation on thymocytes in mice at different power densities. METHODS: The experimental animals were whole body exposed to microwave radiation with frequency of 2,450 MHz, power density of 1, 5, 15 mW/cm(2) respectively 1 h everyday for 30 days. Then the thymus were taken out after the mice were decapitated. Thymus index, morphological characteristics of thymus were examined. The changes of thymus T-cell subgroups, cell cycle progression in thymocytes and cellular apoptosis were detected with flow cytometry (FCM). RESULTS: The body weights of animals in 5, 15 mW/cm(2) irradiation groups [(28.10 +/- 1.46), (27.50 +/- 2.52) g] were lower than that of the control [(31.95 +/- 2.51) g] (P < 0.05). Pathological observation showed dark red piece of nucleus, some nuclei inclined to one side, slight increase in hassall body. The expressions of CD8 in 5, 15 mW/cm(2) irradiation groups (29.14% +/- 1.68%, 29.18% +/- 0.81%) were higher than that in control group (26.95% +/- 1.27%) (P < 0.05). The percentages of G(2) + M phase thymocytes in both radiation groups (12.24% +/- 1.82%, 11.19% +/- 1.36%) were lower than that in control group (14.58% +/- 0.64%) (P < 0.01). Thymocytic apoptosis rates in the three experimental groups (7.18% +/- 0.99%, 10.06% +/- 1.58%, 9.45% +/- 0.92%) were higher than that in control (4.25% +/- 1.63%) (P < 0.01), but the evident difference between 5 mW/cm(2) and 15 mW/cm(2) was not found (P > 0.05). CONCLUSION: Sub-chronic microwave exposure (2 450 MHz, 5, 15 mW/cm(2)) could induce thymocyte apoptosis, cause pathological changes in thymus, and affect cell cycle progression, thus may inhibit the immune function of the animal. PMID- 15130440 TI - [Human factors of drivers and traffic accidents]. AB - OBJECTIVE: To find out the relationship of traffic accidents with life events, driving stress, aggressive driving, etc. METHODS: A total of 905 automobile drivers were investigated with life events scale (LES), general driver stress questionnaire, driver behavior inventory and accidents experience, etc. RESULTS: The scores of life events, driving stress, and aggressive driving in accident drivers (21.79 +/- 14.10, 23.81 +/- 11.86, 9.42 +/- 8.25 respectively) were higher than those in nonaccident drivers (16.82 +/- 8.45, 20.09 +/- 10.63, 5.66 +/- 7.54) (P < 0.01). The number of vehicle accidents was significantly correlated with the scores of LES, driving stress, driver's aggressive behavior, drowsy driving, weekly time of driving, drinking index (P < 0.05). A logistic analysis (Forward:LR) showed that the driving hour, drowsy driving, aggressive driving, and drinking index were closely related with traffic accidents (P < 0.01). CONCLUSION: Life events, driving stress and aggressive driving of the drivers are important factors related to traffic accidents. PMID- 15130441 TI - [Protective effects of heat shock response on circulatory collapse induced by hyperthermia]. AB - OBJECTIVE: To investigate the protective effects and mechanism of heat shock response (HSR) on circulatory collapse induced by hyperthermia. METHODS: Two experiments were carried out: (1) Protective effects of HSR. Rats were divided into 2 groups: heat shock (HS) group, sham control (SC) group. After HS group was pretreated with heat shock and recovered for 20 h at room temperature, both groups were exposed to heat till death, and blood pressure, electrocardiogram were measured continuously during exposure. Mean arterial pressure (MAP), survival time etc were acquired through Chart software. (2) Mechanism of effects. Rats were divided into 3 groups: HS group, SC group and normal control (NC) group. The treatment in HS and SC groups was identical with that in the first experiment, but it would be terminated at 73 min after heat exposure. Systolic pressure (Ps), diastolic pressure (Pd) etc were recorded and content of NO and HSP70 in myocardium were measured. RESULTS: (1) The survival time in HS group [(102.3 +/- 11.4) min] was longer than that in SC group [(87.9 +/- 7.7) min] and shock revealed later (P < 0.01); (2) During early heat exposure MAP in HS group was not different from that in SC group, but after 60 min MAP in HS group were higher than that in SC group; (3) MAP, Ps, Pd, HR and HSP70 in HS group were significantly higher but content of NO was lower than those in SC group (P < 0.01, P < 0.05). CONCLUSION: HSR may induce upregulation of HSP70 and inhibit excessive production of NO in myocardium, thus result in relief of circulatory collapse induced by hyperthermia. PMID- 15130442 TI - [Appraisal of occupational stress and its influential factors in nurses]. AB - OBJECTIVE: To assess the occupational stress and its influential factors in nurses. METHODS: A test of occupational stress, its influential factors, work ability were carried out for 248 nurses and 319 controls with revised occupational stress inventory (OSI-R) and work ability index (WAI). RESULTS: The scores of personal cope resource (131.266 +/- 17.176) and work ability index (32.581 +/- 3.158) in nurse group were significantly higher than those in control group (126.931 +/- 19.108, 31.840 +/- 4.069) (P < 0.05). The main occupational stressors scores (role insufficiency, role clash, and responsibility) in nurses were higher than those in controls (P < 0.05). The stress response of interpersonal relationship in nurses was also higher. The items of personal cope resource, such as recreation, self-care and social support of nurses were superior to those of controls (P < 0.05). Stress response was positively correlated with occupational role (r = 0.512, P < 0.01), and negatively correlated with the personal cope resource (r = -0.475, P < 0.01). The primary influential factors of personal stress were recreation, social support, rational conduct, role insufficiency, role clash, responsibility, and poor work environment. CONCLUSION: To strengthen social support, to improve work condition for nurses, so as to reduce the occupational stress and to enhance the work ability of nurses are important task in occupational health field. PMID- 15130443 TI - [Monitoring indexes for early renal injury in the workers exposed to mercury]. AB - OBJECTIVE: To study the diagnostic method for early renal injury in the workers exposed to mercury (Hg). METHODS: The contents of urinary Hg were determined by chemical method. Urinary microalbumin (mALB), beta(2)-microglodulin (beta(2)-MG) and retinol binding protein (RBP) levels were measured with total quantitative enzyme immunoassay. The activities of urinary N-acetyl-beta-D-glucosaminidase (NAG) and gamma-glutamyl transferase (gamma-GT) were determined by rate methods. Urinary creatinine (Cr) was measured by using picric acid method. RESULTS: The levels of urinary BRP, beta(2)-MG, NAG and gamma-GT in exposed workers [(439.7 +/ 201.4), (141.4 +/- 56.3) micro g/g Cr and (12.3 +/- 5.7), (60.3 +/- 18.5) U/g Cr respectively] were significantly higher than those in controls (P < 0.05, P < 0.01). The levels were increased gradually with the increasing contents of urinary Hg. The positive detection rate for single or two combined indexes was rather lower whereas that for 4 combined indexes was as high as 85.5%. A positive correlation was noted between the contents of urinary Hg and urinary BRP, beta(2) MG, NAG and gamma-GT (r: 0.466, 0.379, 0.323, 0.311, P < 0.05). Urinary RBP was correlated to urinary beta(2)-MG, NAG and gamma-GT (r: 0.362, 0.354, 0.332, P < 0.05). CONCLUSION: Combined detection of urinary RBP, beta(2)-MG, NAG and gamma GT is a sensitive method for the diagnosis of early renal injury in the workers exposed to Hg. PMID- 15130444 TI - [Modeling of acute respiratory distress syndrome in canine after inhalation of perfluoroisobutylene and preliminary study on mechanisms of injury]. AB - OBJECTIVE: To establish of acute respiratory distress syndrome (ARDS) model in canine after inhalation of perfluoroisobutylene (PFIB), and to observe the progressing of lung injury, and to study the mechanisms of injury. METHODS: A device of inhalation of PFIB for canine was made. The concentration of PFIB was 0.30 - 0.32 mg/L. Serum IL-6 and IL-8 were dynamically measured. Clinical manifestations, pathology of organs in canine were observed. RESULTS: (1) During inhalation, the concentration of PFIB remained stable; (2) After inhalation, blood arterial oxygen partial pressure fell gradually, and eventually met the criteria for diagnosing ARDS; (3) The level of IL-8 in serum rises significantly after inhalation (P < 0.05), whereas that of IL-6 was not obviously altered (P > 0.05); (4) Within 6 hours after inhalation, no abnormality in canine was observed, but afterwards symptoms gradually appeared, and typical breath of ARDS, such as high frequency and lower level could be seen in later phase; (5) Pathological examination showed severe congestion, edema and atelectasis in most part of both lungs, and signs of anoxia in other organs. CONCLUSIONS: (1) The device designed is capable of ensuring control of inhalation of PFIB; (2) Exposure to PFIB for 30 mins, canines all met the criteria for diagnosing ARDS 22 hours after inhalation, therefore the modeling is successful; (3) PFIB specifically damages the lung by causing excessive inflammation. PMID- 15130445 TI - [Effects of hearing susceptibility for noise induced hypertension in fertilizer manufacture workers]. PMID- 15130446 TI - [A study on the occupational stress and its influence factors to commercial workers in a market]. PMID- 15130447 TI - [Combined effect of heat and noise on plasma angiotension II and adreomedullin content in pilots]. PMID- 15130448 TI - [Effects of salute sound on blood GSH-Px and plasma MDA in salute soldiers]. PMID- 15130449 TI - [The unbalance of anti-oxidation enzyme system and lipid peroxidation in acute high altitude sickness]. PMID- 15130450 TI - [The relativity between bone marrow mononuclear cells apoptosis and peripheral lymphocyte micronucleus in workers exposed to benzene]. PMID- 15130451 TI - [An experimental study on effects of local vibration on the serum concentration of tumor necrosis factor in rabbits]. PMID- 15130452 TI - [Treatment of 18 patients with acute tetramine poisoning]. PMID- 15130453 TI - [Some problems in treatment of tetramine poisoning]. PMID- 15130454 TI - [Progress in studies on hand-arm vibration disease in China]. PMID- 15130455 TI - [Progress in studies on the oxidative metabolism of xenobiotics mediated by lipoxygenase from extrahepatic tissues]. PMID- 15130456 TI - [Risk factors of occupational hearing loss]. PMID- 15130457 TI - [Determination of arsenic in urine by hydride generation-atomic fluorescence spectroscopy]. PMID- 15130458 TI - [Probe into the definition and hygienic standard of working in heat environment]. PMID- 15130459 TI - [Analysis of the electrocardiograms for 5,184 drivers in Xintai city]. PMID- 15130460 TI - [A view of the occupational health and safety management in Australia]. PMID- 15130461 TI - The clinical and cost-effectiveness of anakinra for the treatment of rheumatoid arthritis in adults: a systematic review and economic analysis. AB - OBJECTIVES: To review the evidence of the clinical and cost-effectiveness of anakinra, an interleukin-1 receptor antagonist (IL-1Ra), for the treatment of rheumatoid arthritis (RA) in adults. DATA SOURCES: Electronic bibliographic databases. Scrip, Food and Drug Administration (FDA) submissions for new drug applications, European Agency for the Evaluation of Medicinal Products (EMEA) reports and the pharmaceutical company submission to the National Institute for Clinical Excellence. REVIEW METHODS: Studies were identified that included randomised controlled trials (RCTs) or economic evaluations of anakinra in adult patients with RA. Existing health economic reviews were also assessed. Data were extracted and quality assessed using a structured approach. The Birmingham Rheumatoid Arthritis Model (BRAM) was used to compare disease-modifying antirheumatic drug (DMARD) sequences, chosen to reflect current clinical practice, with and without anakinra, at different points in the DMARD sequence. RESULTS: Five high-quality RCTs of anakinra in adult patients with RA, involving a total of 2905 patients, of whom 2146 received anakinra, were identified. The results of the clinical trials were consistent with clinical benefit (compared with placebo) as measured by American College of Rheumatology (ACR) composite response rate at 6 months. Variation in response rate was seen across the trials, which is likely to be a reflection of the size of the trials and the wide range of doses evaluated. Consistent benefit was seen at the higher dose evaluated. Benefit was evident both with monotherapy and when used in combination with methotrexate. Data on the efficacy end-points evaluated in a large pragmatic safety study have not been made available, which is of concern. Anakinra treatment was associated with a high incidence of injection-site reactions. Serious adverse events were infrequent, but longer term follow-up is required. No fully published economic evaluations of anakinra in patients with RA were identified. The BRAM gives a base-case estimate of the incremental cost effectiveness ratio (ICER) of anakinra of 106,000 pounds to 604,000 pounds/quality-adjusted life-year (QALY). In the sensitivity analyses substantial variations were made in key parameters and ICERs were shown to be responsive. However, ICERs did not drop below 50,000 pounds/QALY in any univariate sensitivity analysis. CONCLUSIONS: Anakinra can be considered modestly effective in the treatment of RA based on ACR response, although no conclusion can currently be made on the effect of treatment on disease progression. Adjusted indirect comparison suggests that anakinra may be significantly less effective at relieving the clinical signs and symptoms of RA, as measured by the ACR response criteria, than tumour necrosis factor (TNF) inhibitors all used in combination with methotrexate, although these results should be interpreted with caution. The BRAM produces an ICER for anakinra substantially higher than those for infliximab and etanercept. However, patients may respond to anakinra when they have not responded to other TNF inhibitors, as these agents have a different mechanism of action. Thus, anakinra may produce a clinically significant and important improvement in some patients that they could not otherwise have achieved. Further research would be valuable in the following areas: RCTs to evaluate the efficacy, safety and cost of anakinra over the longer term; comparative trials of anakinra with other DMARDs and biological modifiers; assessment of the role of anakinra in the treatment of patients who have failed to achieve a benefit while taking infliximab or etanercept; assessment on the impact of DMARDs and anakinra on joint replacement, mortality and quality of life; controlled clinical trials of combination therapy with two anticytokines; investigations into newer biological therapies; and the utility of radiographic outcomes in clinical trials of RA. PMID- 15130462 TI - Does early imaging influence management and improve outcome in patients with low back pain? A pragmatic randomised controlled trial. AB - OBJECTIVES: To establish whether the early use of sophisticated imaging techniques influences the clinical management and outcome of patients with low back pain (LBP) and whether it is cost-effective. DESIGN: A pragmatic multicentre randomised controlled trial using a standard two parallel group approach incorporating an economic evaluation. For a subgroup of trial participants, a controlled 'before and after' approach was used to assess the impact of 'early imaging' on clinicians' diagnostic and therapeutic confidence. SETTING AND PARTICIPANTS: A total of 782 participants who had been referred by their general practitioner to a consultant orthopaedic specialist or neurosurgeon because of symptomatic lumbar spine disorders. The study included 14 hospitals in Scotland and one in England over a 24-month period. RESULTS: Participants in both groups reported an improvement in health status at 8 and 24 months with the 'early imaging' group having statistically significantly better outcome. Other than the proportion of participants receiving imaging (90% versus 30%), there were few differences between the groups in the management received throughout the 24-month follow-up. The total number of outpatient consultations in the two groups was similar although more people in the 'early imaging' group had return outpatient appointments during the 8-month follow-up. Clinicians' diagnostic confidence, between trial entry and follow-up, increased significantly for both groups with a greater increase in the 'early imaging' group. The cost of imaging was the main determinant of the difference in total costs between the groups and it was estimated that 'early imaging' could provide an additional 0.07 quality-adjusted life-years (QALYs), at an additional average cost of 61 British pounds over the 24-month follow-up. Using non-imputed costs and QALYs but adjusted for baseline differences in EQ-5D score, the mean incremental cost per QALY of 'early imaging' was 870 British pounds. The results were sensitive to the costs of imaging and the confidence intervals surrounding estimates of average costs and QALYs. CONCLUSIONS: The early use of sophisticated imaging does not appear to affect management overall but does result in a slight improvement in clinical outcome at an estimated cost of 870 British pounds per QALY. Imaging was associated with an increase in clinicians' diagnostic confidence, particularly for non-specialists. Further research is required to determine if more rapid referral to sophisticated imaging and secondary care is important in the acute episode and whether the use of imaging would be more beneficial for particular categories of LBP. PMID- 15130463 TI - Curse of the hairpin loop. PMID- 15130464 TI - Secrets of carotenoid binding. PMID- 15130466 TI - Percent sequence identity; the need to be explicit. PMID- 15130465 TI - The structural basis of elasticity in fibrillin-based microfibrils. PMID- 15130467 TI - Crystal structure to 1.7 a of the Escherichia coli pyrimidine nucleoside hydrolase YeiK, a novel candidate for cancer gene therapy. AB - Enzymes with nucleoside hydrolase (NH) activity are crucial for salvaging nucleic acid components in purine auxotrophic protozoan parasites, but are also present in prokaryotes and higher eukaryotes. Here we analyze the distribution of genes encoding for putative NH proteins and characterize the yeiK gene product from Escherichia coli as a pyrimidine-specific NH. The crystal structure of YeiK to 1.7 A defines the structural basis for its substrate specificity and identifies residues involved in the catalytic mechanism that differ from both nonspecific and purine-specific NHs. Large differences in the tetrameric quaternary structure compared to nonspecific protozoan NHs are brought forth by minor differences in the interacting surfaces. The first structural and functional characterization of a nonparasitic, pyrimidine nucleoside-specific NH suggests a possible role for these enzymes in the metabolism of tRNA nucleosides. The high catalytic efficiency of YeiK toward 5-fluorouridine could be exploited for suicide gene therapy in cancer treatment. PMID- 15130468 TI - Structural basis for the specificity, catalysis, and regulation of human uridine cytidine kinase. AB - Uridine-cytidine kinase (UCK) catalyzes the phosphorylation of uridine and cytidine and activates pharmacological ribonucleoside analogs. Here we present the crystal structures of human UCK alone and in complexes with a substrate, cytidine, a feedback inhibitor, CTP or UTP, and with phosphorylation products, CMP and ADP, respectively. Free UCK takes an alpha/beta mononucleotide binding fold and exists as a homotetramer with 222 symmetry. Upon inhibitor binding, one loop region was loosened, causing the UCK tetramer to be distorted. Upon cytidine binding, a large induced fit was observed at the uridine/cytidine binding site, which endows UCK with a strict specificity for pyrimidine ribonucleosides. The first UCK structure provided the structural basis for the specificity, catalysis, and regulation of human uridine-cytidine kinase, which give clues for the design of novel antitumor and antiviral ribonucleoside analogs that inhibit RNA synthesis. PMID- 15130469 TI - Protein regulation of carotenoid binding; gatekeeper and locking amino acid residues in reaction centers of Rhodobacter sphaeroides. AB - X-ray diffraction was used to determine high-resolution structures of the reaction center (RC) complex from the carotenoidless mutant, Rb. sphaeroides R 26.1, without or reconstituted with carotenoids. The results are compared with the structure of the RC from a semiaerobically grown Rb. sphaeroides strain 2.4.1. The investigation reveals the structure of the carotenoid in the different protein preparations, the nature of its binding site, and a plausible mechanism by which the carotenoid is incorporated unidirectionally in its characteristic geometric configuration. The structural data suggest that the accessibility of the carotenoid to the binding site is controlled by a specific "gatekeeper" residue which allows the carotenoid to approach the binding site from only one direction. Carotenoid binding to the protein is secured by hydrogen bonding to a separate "locking" amino acid. The study reveals the specific molecular interactions that control how the carotenoid protects the photosynthetic apparatus against photo-induced oxidative destruction. PMID- 15130470 TI - Structural insights into the catalytic mechanism of Trypanosoma cruzi trans sialidase. AB - Sialidases are a superfamily of sialic-acid-releasing enzymes that are of significant interest due to their implication as virulence factors in the pathogenesis of a number of diseases. However, extensive studies of viral and microbial sialidases have failed to provide a comprehensive picture of their mechanistic properties, in part because the structures of competent enzyme substrate complexes and reaction intermediates have never been described. Here we report these structures for the Trypanosoma cruzi trans-sialidase (TcTS), showing that catalysis by sialidases occurs via a similar mechanism to that of other retaining glycosidases, but with some intriguing differences that may have evolved in response to the substrate structure. PMID- 15130471 TI - Crystal structure of the dachshund homology domain of human SKI. AB - The nuclear protooncoprotein SKI negatively regulates transforming growth factor beta (TGF-beta) signaling in cell growth and differentiation. It directly interacts with the Smads and, by various mechanisms, represses the transcription of TGF-beta-responsive genes. SKI is a multidomain protein that includes a domain bearing high sequence similarity with the retinal determination protein Dachshund (the Dachshund homology domain, DHD). The SKI-DHD has been implicated in SMAD 2/3, N-CoR, SKIP, and PML-RARalpha binding. The 1.65 A crystal structure of the Dachshund homology domain of human SKI is reported here. The SKI-DHD adopts a mixed alpha/beta structure which includes features found in the forkhead/winged helix family of DNA binding proteins, although SKI-DHD is not a DNA binding domain. Residues that form a contiguous surface patch on SKI-DHD are conserved within the Ski/Sno family and with Dachshund, suggesting that this domain may mediate intermolecular interactions common to these proteins. PMID- 15130472 TI - Structure of the neural (N-) cadherin prodomain reveals a cadherin extracellular domain-like fold without adhesive characteristics. AB - Classical cadherins mediate cell-cell adhesion through calcium-dependent homophilic interactions and are activated through cleavage of a prosequence in the late Golgi. We present here the first three-dimensional structure of a classical cadherin prosequence, solved by NMR. The prototypic prosequence of N cadherin consists of an Ig-like domain and an unstructured C-terminal region. The folded part of the prosequence-termed prodomain-has a striking structural resemblance to cadherin "adhesive" domains that could not have been predicted from the amino acid sequence due to low sequence similarities. Our detailed structural and evolutionary analysis revealed that prodomains are distant relatives of cadherin "adhesive" domains but lack all the features known to be important for cadherin-cadherin interactions. The presence of an additional "nonadhesive" domain seems to make it impossible to engage homophilic interactions between cadherins that are necessary to activate adhesion, thus explaining the inactive state of prodomain-bearing cadherins. PMID- 15130473 TI - Molecular basis of box C/D RNA-protein interactions; cocrystal structure of archaeal L7Ae and a box C/D RNA. AB - We have determined and refined a crystal structure of the initial assembly complex of archaeal box C/D sRNPs comprising the Archaeoglobus fulgidus (AF) L7Ae protein and a box C/D RNA. The box C/D RNA forms a classical kink-turn (K-turn) structure and the resulting protein-RNA complex serves as a distinct platform for recruitment of the fibrillarin-Nop5p complex. The cocrystal structure confirms previously proposed secondary structure of the box C/D RNA that includes a protruded U, a UU mismatch, and two sheared tandem GA base pairs. Detailed structural comparisons of the AF L7Ae-box C/D RNA complex with previously determined crystal structures of L7Ae homologs in complex with functionally distinct K-turn RNAs revealed a set of remarkably conserved principles in protein RNA interactions. These analyses provide a structural basis for interpreting the functional roles of the box C/D sequences in directing specific assembly of box C/D sRNPs. PMID- 15130474 TI - The crystal structure of the monomeric reverse transcriptase from Moloney murine leukemia virus. AB - Reverse transcriptases (RTs) are multidomain enzymes of variable architecture that couple both RNA- and DNA-directed DNA polymerase activities with an RNase H activity specific for an RNA:DNA hybrid in order to replicate the single-stranded RNA genome of the retrovirus. Previous structural work has been reported for the heterodimeric HIV-1 and HIV-2 RTs. We now report the first crystal structure of the full-length Moloney murine leukemia virus (MMLV) RT at 3.0 A resolution. The structure reveals a clamp-shaped molecule resulting from the relative positions of the thumb, connection, and RNase H domains that is strikingly different from the HIV-1 RT and provides the first example of a monomeric reverse transcriptase. A comparative analysis with related DNA polymerases suggests a unique trajectory for the template-primer exiting the polymerase active site and provides insights regarding processive DNA synthesis by MMLV RT. PMID- 15130475 TI - Heterogeneity and inaccuracy in protein structures solved by X-ray crystallography. AB - Proteins are dynamic molecules, exhibiting structural heterogeneity in the form of anisotropic motion and discrete conformational substates, often of functional importance. In protein structure determination by X-ray crystallography, the observed diffraction pattern results from the scattering of X-rays by an ensemble of heterogeneous molecules, ordered and oriented by packing in a crystal lattice. The majority of proteins diffract to resolutions where heterogeneity is difficult to identify and model, and are therefore approximated by a single, average conformation with isotropic variance. Here we show that disregarding structural heterogeneity introduces degeneracy into the structure determination process, as many single, isotropic models exist that explain the diffraction data equally well. The large differences among these models imply that the accuracy of crystallographic structures has been widely overestimated. Further, it suggests that analyses that depend on small differences in the relative positions of atoms may be flawed. PMID- 15130476 TI - Crystal structure of the YDR533c S. cerevisiae protein, a class II member of the Hsp31 family. AB - The ORF YDR533c from Saccharomyces cerevisiae codes for a 25.5 kDa protein of unknown biochemical function. Transcriptome analysis of yeast has shown that this gene is activated in response to various stress conditions together with proteins belonging to the heat shock family. In order to clarify its biochemical function, we determined the crystal structure of YDR533c to 1.85 A resolution by the single anomalous diffraction method. The protein possesses an alpha/beta hydrolase fold and a putative Cys-His-Glu catalytic triad common to a large enzyme family containing proteases, amidotransferases, lipases, and esterases. The protein has strong structural resemblance with the E. coli Hsp31 protein and the intracellular protease I from Pyrococcus horikoshii, which are considered class I and class III members of the Hsp31 family, respectively. Detailed structural analysis strongly suggests that the YDR533c protein crystal structure is the first one of a class II member of the Hsp31 family. PMID- 15130477 TI - Crystal structures of apocalmodulin and an apocalmodulin/SK potassium channel gating domain complex. AB - Small conductance Ca2+-activated K+ channels (SK channels) are composed of the pore-forming alpha subunit and calmodulin (CaM). CaM binds to a region of the alpha subunit called the CaM binding domain (CaMBD), located intracellular and immediately C-terminal to the inner helix gate, in either the presence or absence of Ca2+. SK gating occurs when Ca2+ binds the N lobe of CaM thereby transmitting the signal to the attached inner helix gate to open. Here we present crystal structures of apoCaM and apoCaM/SK2 CaMBD complex. Several apoCaM crystal forms with multiple (12) packing environments reveal the same EF hand domain-swapped dimer providing potentially new insight into CaM regulation. The apoCaM/SK2 CaMBD structure, combined with our Ca2+/CaM/CaMBD structure suggests that Ca2+ binding induces folding and dimerization of the CaMBD, which causes large CaMBD-CaM C lobe conformational changes, including a >90 degrees rotation of the region of the CaMBD directly connected to the gate. PMID- 15130478 TI - Structure of the N-terminal domain of Escherichia coli glutamine synthetase adenylyltransferase. AB - We report the crystal structure of the N-terminal domain of Escherichia coli adenylyltransferase that catalyzes the reversible nucleotidylation of glutamine synthetase (GS), a key enzyme in nitrogen assimilation. This domain (AT-N440) catalyzes the deadenylylation and subsequent activation of GS. The structure has been divided into three subdomains, two of which bear some similarity to kanamycin nucleotidyltransferase (KNT). However, the orientation of the two domains in AT-N440 differs from that in KNT. The active site of AT-N440 has been identified on the basis of structural comparisons with KNT, DNA polymerase beta, and polyadenylate polymerase. AT-N440 has a cluster of metal binding residues that are conserved in polbeta-like nucleotidyl transferases. The location of residues conserved in all ATase sequences was found to cluster around the active site. Many of these residues are very likely to play a role in catalysis, substrate binding, or effector binding. PMID- 15130479 TI - Probing the energy landscape of the membrane protein bacteriorhodopsin. AB - The folding and stability of transmembrane proteins is a fundamental and unsolved biological problem. Here, single bacteriorhodopsin molecules were mechanically unfolded from native purple membranes using atomic force microscopy and force spectroscopy. The energy landscape of individual transmembrane alpha helices and polypeptide loops was mapped by monitoring the pulling speed dependence of the unfolding forces and applying Monte Carlo simulations. Single helices formed independently stable units stabilized by a single potential barrier. Mechanical unfolding of the helices was triggered by 3.9-7.7 A extension, while natural unfolding rates were of the order of 10(-3) s(-1). Besides acting as individually stable units, helices associated pairwise, establishing a collective potential barrier. The unfolding pathways of individual proteins reflect distinct pulling speed-dependent unfolding routes in their energy landscapes. These observations support the two-stage model of membrane protein folding in which alpha helices insert into the membrane as stable units and then assemble into the functional protein. PMID- 15130480 TI - The focal adhesion targeting domain of focal adhesion kinase contains a hinge region that modulates tyrosine 926 phosphorylation. AB - The focal adhesion targeting (FAT) domain of focal adhesion kinase (FAK) is critical for recruitment of FAK to focal adhesions and contains tyrosine 926, which, when phosphorylated, binds the SH2 domain of Grb2. Structural studies have shown that the FAT domain is a four-helix bundle that exists as a monomer and a dimer due to domain swapping of helix 1. Here, we report the NMR solution structure of the avian FAT domain, which is similar in overall structure to the X ray crystal structures of monomeric forms of the FAT domain, except that loop 1 is longer and less structured in solution. Residues in this region undergo temperature-dependent exchange broadening and sample aberrant phi and psi angles, which suggests that this region samples multiple conformations. We have also identified a mutant that dimerizes approximately 8 fold more than WT FAT domain and exhibits increased phosphorylation of tyrosine 926 both in vitro and in vivo. PMID- 15130481 TI - Structure of protein L7Ae bound to a K-turn derived from an archaeal box H/ACA sRNA at 1.8 A resolution. AB - The archaeal RNA binding protein L7Ae and its eukaryotic homolog 15.5 kDa/Snu13 recognize K-turns. This structural motif is canonically comprised of two stems (one with tandem A.G base pairs, the other with Watson-Crick pairs) linked by an asymmetric internal loop. L7Ae recognizes conventional K-turns in ribosomal and box C/D RNAs but also binds specifically to some box H/ACA RNAs at terminal stem loops. These have the A.G paired stem, but lack the Watson-Crick stem. The structure of Methanococcus jannaschii L7Ae bound to a symmetric duplex RNA without Watson-Crick stems demonstrates how a binding site for this component of diverse ribonucleoprotein complexes can be constructed with only the A.G stem and the loop. The RNA adopts a functional conformation with the aid of a base triple and tight binding of divalent cations. Comparison with the 15.5 kDa/Snu13-RNA complex structure suggests why the eukaryotic homolog does not recognize terminal stem loop L7Ae binding sites. PMID- 15130482 TI - Cyclin C makes an entry into the cell cycle. AB - From yeast to humans, cell cycle progression is orchestrated by the oscillation of kinase activities associated with cyclins. In an article published recently in Cell, Ren and Rollins investigate mechanisms controlling the G0/G1 transition in quiescent cells and identify new cyclin C/Cdk3 complexes as key regulators of cell cycle reentry in human cells. PMID- 15130483 TI - Mitosis: FEAR pulls them apart. AB - The function of astral microtubules during mitosis is becoming the focus of increasing interest. A report published in this issue of Developmental Cell provides new insight into the complex and subtle regulation of these microtubules during anaphase. PMID- 15130484 TI - Cathepsins as transcriptional activators? AB - One action of proteases is limited cleavage of latent transcription factors, releasing active factors to initiate nuclear signaling. Goulet et al., in a recent Molecular Cell paper, provide evidence that a lysosomal protease, cathepsin L, exists in a previously unsuspected isoform capable of trafficking to the nucleus and activating the CDP/Cux transcription factor. The findings should stimulate new research on the nature of nuclear proteases involved in signaling. PMID- 15130485 TI - Lost in translation gets an oskar. AB - Nuclear history affects the fates of mRNAs in the cytoplasm of cells. Proteins loaded onto mRNAs in the nucleus mark RNAs for subsequent translational regulation, stability, degradation, and subcellular RNA localization. New results show that the Drosophila heterogeneous nuclear ribonucleoprotein (hnRNP) Hrp48 contributes to coordinated RNA localization and translational control in oocytes. PMID- 15130486 TI - Reduce your pelvis in 10000 years or less. AB - While the avid dieter obsesses about reduction of different body parts, permanent reduction of many structures seems to be achieved relatively effortlessly (though much more slowly) throughout evolution. In a recent article in Nature, Shapiro et al. examine one such example, the genetic basis of pelvic reduction in the threespine stickleback fish (2004). They conclude that a regulatory mutation in the Pitx1 gene is responsible for the pelvic reduction. PMID- 15130487 TI - Reduction and compaction in the genome of the apicomplexan parasite Cryptosporidium parvum. AB - The complete genome of the apicomplexan parasite Cryptosporidium parvum reveals many new insights into apicomplexan biology and evolution, as well as the general process of genome reduction in parasites. The genome is globally compacted, but gene loss seems to be focused, in particular in relation to organelles. Massive losses of mitochondrial genes have taken place and there is no evidence of any plastid-related genes, providing a useful tool for examining putative plastid proteins in Plasmodium and other apicomplexans. PMID- 15130488 TI - The Drosophila hnRNPA/B homolog, Hrp48, is specifically required for a distinct step in osk mRNA localization. AB - The Staufen-dependent localization of oskar mRNA to the posterior of the Drosophila oocyte induces the formation of the pole plasm, which contains the abdominal and germline determinants. In a germline clone screen for mutations that disrupt the posterior localization of GFP-Staufen, we isolated three missense alleles in the hnRNPA/B homolog, Hrp48. These mutants specifically abolish osk mRNA localization, without affecting its translational control or splicing, or the localization of bicoid and gurken mRNAs and the organization of the microtubule cytoskeleton. Hrp48 colocalizes with osk mRNA throughout oogenesis, and interacts with its 5' and 3' regulatory regions, suggesting that it binds directly to oskar mRNA to mediate its posterior transport. The hrp48 alleles cause a different oskar mRNA localization defect from other mutants, and disrupt the formation of GFP-Staufen particles. This suggests a new step in the localization pathway, which may correspond to the assembly of Staufen/oskar mRNA transport particles. PMID- 15130489 TI - Hrp48, a Drosophila hnRNPA/B homolog, binds and regulates translation of oskar mRNA. AB - Establishment of the Drosophila embryonic axes provides a striking example of RNA localization as an efficient mechanism for protein targeting within a cell. oskar mRNA encodes the posterior determinant and is essential for germline and abdominal development in the embryo. Tight restriction of Oskar activity to the posterior is achieved by mRNA localization-dependent translational control, whereby unlocalized mRNA is translationally repressed and repression is overcome upon mRNA localization. Here we identify the previously reported oskar RNA binding protein p50 as Hrp48, an abundant Drosophila hnRNP. Analysis of three hrp48 mutant alleles reveals that Hrp48 levels are crucial for polarization of the oocyte during mid-oogenesis. Our data also show that Hrp48, which binds to the 5' and 3' regions of oskar mRNA, plays an important role in restricting Oskar activity to the posterior of the oocyte, by repressing oskar mRNA translation during transport. PMID- 15130490 TI - The transitional ER localization mechanism of Pichia pastoris Sec12. AB - COPII vesicles assemble at ER subdomains called transitional ER (tER) sites, but the mechanism that generates tER sites is unknown. To study tER biogenesis, we analyzed the transmembrane protein Sec12, which initiates COPII vesicle formation. Sec12 is concentrated at discrete tER sites in the budding yeast Pichia pastoris. We find that P. pastoris Sec12 exchanges rapidly between tER sites and the general ER. The tER localization of Sec12 is saturable and is mediated by interaction of the Sec12 cytosolic domain with a partner component. This interaction apparently requires oligomerization of the Sec12 lumenal domain. Redistribution of P. pastoris Sec12 to the general ER does not perturb the localization of downstream tER components, suggesting that Sec12 and other COPII proteins associate with a tER scaffold. These results provide evidence that tER sites form by a network of dynamic associations at the cytosolic face of the ER. PMID- 15130491 TI - Skp2-mediated degradation of p27 regulates progression into mitosis. AB - Although Skp2 has been thought to mediate the degradation of p27 at the G(1)-S transition, Skp2(-/-) cells exhibit accumulation of p27 in S-G(2) phase with overreplication. We demonstrate that Skp2(-/-)p27(-/-) mice do not exhibit the overreplication phenotype, suggesting that p27 accumulation is required for its development. Hepatocytes of Skp2(-/-) mice entered the endoduplication cycle after mitogenic stimulation, whereas this phenotype was not apparent in Skp2(-/ )p27(-/-) mice. Cdc2-associated kinase activity was lower in Skp2(-/-) cells than in wild-type cells, and a reduction in Cdc2 activity was sufficient to induce overreplication. The lack of p27 degradation in G(2) phase in Skp2(-/-) cells may thus result in suppression of Cdc2 activity and consequent inhibition of entry into M phase. These data suggest that p27 proteolysis is necessary for the activation of not only Cdk2 but also Cdc2, and that Skp2 contributes to regulation of G(2)-M progression by mediating the degradation of p27. PMID- 15130492 TI - Deacetylase inhibitors increase muscle cell size by promoting myoblast recruitment and fusion through induction of follistatin. AB - Fusion of undifferentiated myoblasts into multinucleated myotubes is a prerequisite for developmental myogenesis and postnatal muscle growth. We report that deacetylase inhibitors favor the recruitment and fusion of myoblasts into preformed myotubes. Muscle-restricted expression of follistatin is induced by deacetylase inhibitors and mediates myoblast recruitment and fusion into myotubes through a pathway distinct from those utilized by either IGF-1 or IL-4. Blockade of follistatin expression by RNAi-mediated knockdown, functional inactivation with either neutralizing antibodies or the antagonist protein myostatin, render myoblasts refractory to HDAC inhibitors. Muscles from animals treated with the HDAC inhibitor trichostatin A display increased production of follistatin and enhanced expression of markers of regeneration following muscle injury. These data identify follistatin as a central mediator of the fusigenic effects exerted by deacetylase inhibitors on skeletal muscles and establish a rationale for their use to manipulate skeletal myogenesis and promote muscle regeneration. PMID- 15130493 TI - The clonal origin of myocardial cells in different regions of the embryonic mouse heart. AB - When and how cells form and pattern the myocardium is a central issue for heart morphogenesis. Many genes are differentially expressed and function in subsets of myocardial cells. However, the lineage relationships between these cells remain poorly understood. To examine this, we have adopted a retrospective approach in the mouse embryo, based on the use of the laacZ reporter gene, targeted to the alpha-cardiac actin locus. This clonal analysis demonstrates the existence of two lineages that segregate early from a common precursor. The primitive left ventricle and the presumptive outflow tract are derived exclusively from a single lineage. Unexpectedly, all other regions of the heart, including the primitive atria, are colonized by both lineages. These results are not consistent with the prespecification of the cardiac tube as a segmented structure. They are discussed in the context of different heart fields and of the evolution of the heart. PMID- 15130494 TI - Negative regulation of midline vascular development by the notochord. AB - The negative regulation of vascular patterning is one of the least understood processes in vascular biology. In amniotes, blood vessels develop throughout the embryonic disc, except for a midline region surrounding the notochord. Here we show that the notochord is the primary signaling center for the inhibition of vessel formation along the embryonic midline. Notochord ablation in quail embryos results in vascular plexus formation at midline. Implantation of the notochord into paraxial and lateral mesoderm inhibits vessel formation locally. The notochord-expressed BMP antagonists Chordin and Noggin inhibit endothelial cell migration in vitro, and their ectopic expression in vivo results in a local disruption of vessel formation. Conversely, BMP-4 activates endothelial cell migration in vitro, and its ectopic expression along the notochord induces vascular plexus formation at midline. These data indicate an inhibitory role of the notochord in defining an avascular zone at the embryonic midline, in part via BMP antagonism. PMID- 15130495 TI - SLIT2-mediated ROBO2 signaling restricts kidney induction to a single site. AB - Kidney development occurs in a stereotypic position along the body axis. It begins when a single ureteric bud emerges from the nephric duct in response to GDNF secreted by the adjacent nephrogenic mesenchyme. Posterior restriction of Gdnf expression is considered critical for correct positioning of ureteric bud development. Here we show that mouse mutants lacking either SLIT2 or its receptor ROBO2, molecules known primarily for their function in axon guidance and cell migration, develop supernumerary ureteric buds that remain inappropriately connected to the nephric duct, and that the SLIT2/ROBO2 signal is transduced in the nephrogenic mesenchyme. Furthermore, we show that Gdnf expression is inappropriately maintained in anterior nephrogenic mesenchyme in these mutants. Thus our data identify an intercellular signaling system that restricts, directly or indirectly, the extent of the Gdnf expression domain, thereby precisely positioning the site of kidney induction. PMID- 15130496 TI - Renin cells are precursors for multiple cell types that switch to the renin phenotype when homeostasis is threatened. AB - Renin-synthesizing cells are crucial in the regulation of blood pressure and fluid-electrolyte homeostasis. Adult mammals subjected to manipulations that threaten homeostasis increase circulating renin by increasing the number of renin expressing/-releasing cells. We hypothesize that the ability of adult cells to synthesize renin does not occur randomly in any cell type, depending instead on the cell's lineage. To determine the fate of renin-expressing cells, we generated knockin mice expressing cre recombinase in renin-expressing cells and crossed them with reporter mice. Results show that renin-expressing cells are precursors for a variety of cells that differentiate into non-renin-expressing cells such as smooth-muscle, epithelial, mesangial, and extrarenal cells. In the kidney, these cells retain the capability to synthesize renin when additional hormone is required to reestablish homeostasis: specific subpopulations of apparently differentiated cells are "held in reserve" to respond (repeatedly) by de differentiating and expressing renin in response to stress, and re differentiating when the crisis passes. PMID- 15130497 TI - A role for the FEAR pathway in nuclear positioning during anaphase. AB - In budding yeast, cells lacking separase function exit mitosis with an undivided nucleus localized to the daughter cell. Here we show that the inability to separate sister chromatids per se is not sufficient to cause the daughter preference. Rather, separase affects nuclear positioning as part of the Cdc14 early anaphase release (FEAR) pathway. The role of the FEAR pathway in nuclear positioning is exerted during anaphase and is not shared by the mitotic exit network. We find that the nuclear segregation defect in FEAR mutants does not stem from nonfunctional spindle poles or the absence of cytoplasmic microtubules. Instead, the concomitant inactivation of sister chromatid separation and the FEAR pathway uncovered a mother-directed force in anaphase that was previously masked by the elongating spindle. We propose that at anaphase onset, the FEAR pathway activates cytoplasmic microtubule-associated forces that facilitate chromosome segregation to the mother cell. PMID- 15130499 TI - DC-virus interplay: a double edged sword. AB - Myeloid and plasmacytoid dendritic cells, a family of professional antigen presenting cells, are crucial in generating and maintaining anti-viral immunity. Many viruses have evolved to avoid, subvert, and even counterattack them. In this article, we focus on the tuning of innate and adaptive responses induced by human dendritic cells, and on the inhibition of their functions by viruses of medical significance. A constant "tug of war" goes on between dendritic cells and viruses and a main dendritic cell countermeasure is cross-presentation/priming. PMID- 15130500 TI - Anatomical features of anti-viral immunity in the respiratory tract. AB - The mucosal surfaces of the lungs are a major portal of entry for virus infections and there are urgent needs for new vaccines that promote effective pulmonary immunity. However, we have only a rudimentary understanding of the requirements for effective cellular immunity in the respiratory tract. Recent studies have revealed that specialized cellular immune responses and lymphoid tissues are involved in the protection of distinct anatomical microenvironments of the respiratory tract, such as the large airways of the nose and the alveolar airspaces. This review discusses some of the anatomical features of anti-viral immunity in the respiratory tract including the role of local lymphoid tissues and the relationship between effector and memory T cells in the airways, the lung parenchyma, and lymphoid organs. PMID- 15130501 TI - CD4 T cell responses to influenza infection. AB - Immune responses to viral infections involve a complex orchestration between innate signals and adaptive responses of specific T and B cells. Anti-viral CD4 cells can direct CD8 responses by secreting a Type 1 panel of cytokines including IFN-gamma, IL-2 and TNF-alpha and can drive B cell production of IgG2a to neutralize infective viral particles. This review will focus specifically on the role of CD4 cells in the immune response to influenza, an acute, localized respiratory viral infection. We suggest that CD4 cells act as direct effectors in protection against influenza, may contribute to immunopathology and generate functionally distinct memory subsets. PMID- 15130502 TI - Characterization of CD8+ T cell repertoire diversity and persistence in the influenza A virus model of localized, transient infection. AB - Influenza virus infection of C57BL/6 mice provides a well-characterized model for the study of acute CD8(+) T cell responses and for the analysis of memory in the absence of antigen persistence. The advent of tetramer reagents and intracellular cytokine staining, coupled with techniques such as single cell RT-PCR and influenza reverse genetics, has enabled the detailed molecular dissection of different epitope-specific primary, memory and secondary immune CD8(+) T cell responses. The approach offers novel insights into the factors determining the selection of immune repertoires, and their functional consequences for CD8(+) T cell-mediated immunity. PMID- 15130503 TI - Role of T cell costimulation in anti-viral immunity. AB - Members of both the CD28 and TNFR families can have costimulatory roles in T cell activation. Gene targeted mice as well as in vivo blocking experiments have established distinct roles for CD28/B7; ICOS/ICOSL; CD27/CD70; 4-1BB/4-1BBL and OX40/OX40L during viral infection. Many issues remain to be addressed, including the timing and location of the interactions, the possibility of partial redundancy between related family members and the molecular basis for the specific phenotypes observed in the different gene targeted mice. PMID- 15130504 TI - Immunological memory in humans. AB - Specificity and memory are the defining characteristics of adaptive immune responses. Vaccines are predicated on the existence of immune memory, and the robustness of immune memory is a primary determinant of vaccine efficacy. How is immune memory maintained? Much progress has been made in this area over the past several years, and new human studies have added key insights into the longevity of B and T cell immune memory in the absence of antigen. PMID- 15130505 TI - Lessons from the study of T-cell differentiation in persistent human virus infection. AB - Confusion surrounds the current classification of memory and effector T-cell subsets and there is a lack of consistency in the use of these terms between human and murine studies. The development of peptide-HLA tetrameric complexes ("tetramers") that accurately identify virus-specific T cells and can be used with a range of cell surface and intra-cellular markers has provided further insights in our understanding of the process of T-cell differentiation, or post thymic development. We propose that T-cell differentiation subsets in human viral infection should be regarded as distinct from the current definitions of memory and effector cells; further work is needed to reveal the role of the differentiation process in anti-viral immunity. PMID- 15130506 TI - A visualisation concept of dynamic signalling networks. AB - Cellular processes and pathways are typically delineated in arrow diagrams to depict the relationship between the individual genes or proteins. These simplifications neglect the complexity and non-linearity of cellular regulatory networks. In this review, a three-dimensional (3D) visualisation model is presented that can be helpful in understanding the non-linear behaviour and limited predictability of regulatory networks, as well as their organisation, stability, attractor states, and evolution. A better understanding of the current views of large, complex scale-free networks can assist in planning and interpretation of high throughput microarray experiments. PMID- 15130507 TI - The role of 17 beta-hydroxysteroid dehydrogenases. AB - The biological activity of steroid hormones is regulated at the pre-receptor level by several enzymes including 17 beta-hydroxysteroid dehydrogenases (17 beta -HSD). The latter are present in many microorganisms, invertebrates and vertebrates. Dysfunctions in human 17 beta-hydroxysteroid dehydrogenases result in disorders of biology of reproduction and neuronal diseases, the enzymes are also involved in the pathogenesis of various cancers. 17 beta-hydroxysteroid dehydrogenases reveal a remarkable multifunctionality being able to modulate concentrations not only of steroids but as well of fatty and bile acids. Current knowledge on genetics, biochemistry and medical implications is presented in this review. PMID- 15130508 TI - Crustacean retinoid-X receptor isoforms: distinctive DNA binding and receptor receptor interaction with a cognate ecdysteroid receptor. AB - We have identified cDNA clones that encode homologs of the ecdysteroid receptor (EcR) and retinoid-X receptor (RXR)/USP classes of nuclear receptors from the fiddler crab Uca pugilator (UpEcR and UpRXR). Several UpRXR cDNA splicing variants were found in coding regions that could potentially influence function. A five-amino acid (aa) insertion/deletion is located in the "T" box in the hinge region. Another 33-aa insertion/deletion is found inside the ligand-binding domain (LBD), between helix 1 and helix 3. Ribonuclease protection assays (RPA) showed that four UpRXR transcripts [UpRXR(+5+33), UpRXR(-5+33), UpRXR(+5-33) and UpRXR(-5-33)] were present in regenerating limb buds. UpRXR(-5+33) was the most abundant transcript present in regenerating limb buds in both early blastema and late premolt growth stages. Expression vectors for these UpRXR variants and UpEcR were constructed, and the proteins expressed in E. coli and in vitro expression systems. The expressed crab nuclear receptors were then characterized by electrophoretic mobility shift assay (EMSA) and glutathione S-transferase (GST) pull down experiments. EMSA results showed that UpEcR/UpRXR(-5+33) heterocomplexes bound with a series of hormone response elements (HREs) including eip28/29, IRper-1, DR-4, and IRhsp-1 with appreciable affinity. Competition EMSA also showed that the affinity decreased as sequence composition deviated from a perfect consensus element. Binding to IRper-1 HREs occurred only if the heterodimer partner UpRXR contained the 33-aa LBD insertion. UpRXR lacking both the 5-aa and 33-aa insertion bound to a DR-1G HRE in the absence of UpEcR. The results of GST-pull down experiments showed that UpEcR interacted only with UpRXR variants containing the 33-aa insertion, and not with those lacking the 33-aa insertion. These in vitro receptor protein-DNA and receptor protein-protein interactions occurred in the absence of hormone (20-hydroxyecdysone and 9-cis retinoid acid, 9-cis RA). Transactivation studies using a hybrid UpEcR ligand binding domain construct and UpRXR (+/-33) ligand-binding domain constructs also showed that the 33-aa insertion was indispensable in mediating ecdysteroid stimulated transactivation. PMID- 15130509 TI - Receptor interacting protein 140 as a thyroid hormone-dependent, negative co regulator for the induction of cellular retinoic acid binding protein I gene. AB - Over-expression of receptor interacting protein 140 (RIP140) suppressed thyroid hormone (T3) induction of cellular retinoic acid binding I protein (CRABPI) gene in P19 embryonal carcinoma cells. CRABPI induction by T3 is mediated by a direct repeat four-element bound by T3 receptor (T3R) and retinoid receptor X (RXR). Three receptor-interacting domains (RIDs) in RIP140 mediate its interaction with T3R: one constitutive RID within the amino terminus, and two T3-dependent RIDs in the central portion and the carboxyl terminus. In co-immunoprecipitation and chromatin immunoprecipitation assays, RIP140 formed complexes with T3R/RXR in solution and on the endogenous target, the CRABPI promoter. T3 treatment resulted in elevated histone acetylation of the endogenous CRABPI gene promoter, but simultaneous expression of RIP140 resulted in significantly reduced histone acetylation of this promoter, primarily through the recruitment of HDAC4. This study presents the first evidence that over-expressed RIP140 acts as a T3 dependent negative co-regulator for T3 induction of the endogenous CRABPI gene in P19 cells. PMID- 15130510 TI - An in vitro bioassay to determine individual sensitivity to glucocorticoids: induction of FKBP51 mRNA in peripheral blood mononuclear cells. AB - Individual variation in sensitivity to glucocorticoids (GCs) poses a dilemma to the clinician. Currently available assays to determine individual sensitivity to GCs either seem imprecise, or they are based on mitogen-activated lymphocytes, although mitogens themselves may affect cellular GC sensitivity. To avoid these disadvantages, we developed an assay based on the GC-induced accumulation of the 51kDa FK506 binding protein (FKBP51) mRNA in unstimulated peripheral blood mononuclear cells (PBMC), measured using real time PCR. Of several family members tested, only FKBP51 transcript levels showed to be GC-inducible. Furthermore, our bioassay was not affected by progesterone, estradiol, and testosterone. Immunological stimulation of PBMC using tetanus toxoid did not affect bioassay results, and isolated T- and B-lymphocytes showed a similar response to GC stimulation. The intra- and inter-assay variations were 10.6 and 15.9%, respectively. Our bioassay confirms previous reports that a wide variation in GC sensitivity exists in the normal population, yet is able to clearly discriminate a patient with familial GC hyposensitivity from controls. Our bioassay may be suitable to assess altered individual GC sensitivity, and the small amount of PBMC needed for a determination makes this assay easily applicable in a pediatric setting. PMID- 15130511 TI - Dehydroepiandrosterone up-regulates resistin gene expression in white adipose tissue. AB - Dehydroepiandrosterone (DHEA), the most abundant steroid hormone in human blood, is considered to be one of fat-reducing hormones. However, the molecular mechanisms underlying DHEA mode of action in obesity has not been fully clarified. The pivotal role in the maintenance of cellular lipid and energy balance is played by peroxisome proliferator-activated receptor alpha (PPARalpha) which acts as transcriptional activator of numerous genes encoding enzymes involved in fatty acid catabolism. Lately published papers suggest that resistin, a low molecular-weight protein produced by adipose tissue, may act as an inhibitor of adipocyte differentiation and could regulate adipose tissue mass. Recent studies have established that the promoter region of the resistin gene contains several putative PPAR response elements. Since DHEA has been characterized as a peroxisome proliferator able to induce hepatic genes through PPARalpha, we hypothesised that DHEA might affect PPARalpha and, subsequently, resistin gene expression in adipose tissue. In order to test this hypothesis, an experiment was performed comparing PPARalpha and resistin gene expression in white adipose tissue (WAT) of male Wistar rats fed standard or DHEA-supplemented (0.6% (w/w)) diet for 2 weeks. DHEA administration to the rats induced PPARalpha and resistin gene expression in WAT (3- and 2.25-fold, respectively; as determined by real-time reverse transcription-polymerase chain reaction (RT PCR)); reduced body weight, epididymal adipose tissue mass and decreased serum leptin levels. We propose that DHEA may impact on the transcription of resistin gene through a mechanism involving PPARalpha and that an elevated resistin level may lead to an inhibition of adipogenesis and a decrease in adipose tissue mass. PMID- 15130512 TI - Targeting estrogen responsive elements (EREs): design of potent transactivators for ERE-containing genes. AB - The estrogen hormone (E2) plays an important role in the physiology and pathophysiology of target tissues. The effects of E2 are conveyed by the estrogen receptors (ER) alpha and beta. The E2-ER complex mediates an array of genomic and non-genomic events that orchestrate the expression of a number of genes involved in the regulation of cell proliferation and differentiation. The interaction of with the regulatory DNA sequence, estrogen responsive element (ERE), of each responsive gene constitutes a critical genomic signaling pathway. However, the relative importance of ERE-dependent E2-ER signaling in cell proliferation remains to be elucidated. To address this issue, we engineered ERE-binding activators (EBAs) that specifically and potently regulate ERE-containing genes. The modular nature of ER allowed us to initially design a monomeric ERE-binding module by genetically joining two DNA-binding domains with the hinge domain. Integration of strong activation domains from other transcription factors into this module generated constitutively active EBAs. These transactivators robustly induced the expression of only ERE-containing promoter constructs in transfected cells independent of ligand, dimerization, ER-subtype and -status. Moreover, EBAs altered cell cycle progression in breast cancer cell lines in a manner similar to E2-ER. These results demonstrate the importance of ERE-containing genes in the regulation of cell proliferation. These novel ERE-binding transregulators could also be a basis for the targeted regulation of ERE-containing genes, the identification of estrogen responsive gene networks, and the development of alternative/complementary therapeutic approaches for estrogen target tissue cancers. PMID- 15130513 TI - Hormonal regulation of human trophoblast differentiation: a possible role for 17beta-estradiol and GnRH. AB - We have examined the role of 17beta-estradiol and gonadotropin releasing hormone (GnRH) in the regulation of functional differentiation in human trophoblasts. In contrast to its recognized functions as a proliferation-promoting hormone in a variety of cell types, we found that 17beta-estradiol induced terminal differentiation in human trophoblastic cells, and that this event was estrogen receptor-mediated. This process involved a loss in expression of Cyclins A2 and E, and a coincident increase in p27(Kip1). The anti-proliferative effects of 17beta-estradiol were annulled by specific transforming growth factor-beta 1 (TGFbeta1)-neutralizing antibody, suggesting that 17beta-estradiol may mediate its growth-inhibitory actions, through TGFbeta1 activity. Following exposure to Buserelin, cultured human trophoblastic cells stopped proliferating and formed functionally mature syncytiotrophoblasts. This differentiation event, that involved a drastic loss in expression of proliferating-cell-nuclear-antigen, could be blocked by Cetrorelix, suggesting the involvement of functional GnRH receptors. Preliminary studies on the characterization of the human placental GnRH receptor, indicate the presence of multiple receptor isoforms across human gestation. PMID- 15130514 TI - The early gene E74B isoform is a transcriptional activator of the ecdysteroid regulatory hierarchy in mosquito vitellogenesis. AB - In the mosquito Aedes aegypti, blood feeding activates vitellogenesis that involves yolk protein precursor (YPP) genes in an insect metabolic tissue, the fat body. Vitellogenesis is regulated by the 20-hydroxyecdysone (20E) regulatory hierarchy, in which the Ets-domain protein E74 is a key transcriptional regulator. The mosquito AaE74 gene encodes two isoforms-AaE74A and AaE74B. Both AaE74 isoforms are 20E-inducible early gene products. AaE74B reaches its maximal expression at 10(-7)M of 20E, while AaE74A requires 10(-6)M of 20E, a concentration at which the YPP genes reach their maximal induction level. In transfection assay, AaE74B is capable of activating a reporter construct containing E74-response elements, while expression of AaE74A has no effect on the basal levels of the reporter. The AaE74B binding activity is present in the fat body nuclei only during active vitellogenesis. Taken together, our findings demonstrate that AaE74B isoform plays the role of a transcriptional activator during vitellogenesis. PMID- 15130515 TI - Differential expression of largemouth bass (Micropterus salmoides) estrogen receptor isotypes alpha, beta, and gamma by estradiol. AB - The expression levels of three estrogen receptor (ER) isotypes alpha, beta, and gamma were quantified in female largemouth bass (Micropterus salmoides) (LMB) liver, ovary, brain, and pituitary tissues. ER alpha and beta expression predominated in the liver, while ERs beta and gamma predominated in the other tissues. Temporally in females, ER alpha was highly up-regulated, ER gamma was slightly up-regulated, and ER beta levels remained unchanged in the liver when plasma 17-beta estradiol (E2) and vitellogenin (Vtg) levels were elevated in the spring. In ovarian tissue from these same fish, all three ERs were maximally expressed in the fall, during early oocyte development and prior to peak plasma E2 levels. When males were injected with E2, ER alpha was highly inducible, ER gamma was moderately up-regulated, and ER beta levels were not affected. None of the ER isotypes were induced by E2 in gonadal tissues. These results combined suggest that the ERs themselves are not regulated in the same manner by E2, and furthermore, do not contribute equally to the transcriptional regulation of genes involved in fish reproduction such as Vtg. PMID- 15130516 TI - Roles of fibroblast growth factor 10 (Fgf10) in adipogenesis in vivo. AB - The development of white adipose tissue (WAT) of Fgf10-/- mouse embryos was greatly impaired. Here, we examined the mechanism of Fgf10 action in adipogenesis in vivo. The proliferative activity in the WAT of Fgf10-/- embryos was greatly decreased. We also examined the expression of transcription factors, C/EBPbeta, C/EBPalpha and PPARgamma, that are important for adipogenesis. Although the expression of C/EBPbeta and PPARgamma in the WAT of Fgf10-/- embryos was greatly decreased, the expression of C/EBPalpha was essentially unchanged. Therefore, we examined their expression in the WAT of C/EBPalpha-/- embryos. Although the expression of C/EBPbeta and PPARgamma in the WAT was greatly decreased, the expression of Fgf10 was essentially unchanged. As these results in vivo appeared to be contradictory to a transcriptional cascade model in vitro that C/EBPbeta induces the expression of PPARgamma and C/EBPalpha reported, we also examined their expression in the WAT of wild type embryos at different developmental stages. The expression of Fgf10 and C/EBPalpha was followed by that of C/EBPbeta and PPARgamma. The present findings indicate that Fgf10 but not C/EBPalpha is required for the proliferation of preadipocytes. In contrast, both Fgf10 and C/EBPalpha acting synergistically in separate, parallel pathways are required for the differentiation. Unexpectedly, the transcriptional cascade of adipogenesis in vivo described here is distinct from the cascade in vitro previously reported. PMID- 15130517 TI - Human endometrium and decidua express follistatin-related gene (FLRG) mRNA and peptide. AB - Activin-A is expressed by human endometrium, and the actions are counteracted by follistatin, its binding protein. We evaluated the endometrial mRNA and peptide expression of follistatin-related gene (FLRG), a protein that binds activin-A, preventing its interaction. By reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry, FLRG expression was evaluated in tissues collected at early proliferative (EP; n = 8) and late proliferative (LP; n = 8); early secretory (ES; n = 9) and late secretory (LS; n = 10); and in pregnancy, maternal decidua (MD; n = 12). FLRG mRNA was expressed by all samples, and semi quantitative analysis showed that FLRG expression was significantly ( P < 0.001) higher in MD. FLRG was strongly immunolocalized in epithelial cells of glands and vessel walls (cytoplasma and nucleus), but only in the stromal cells nucleus. In MD, FLRG immunostaining was found in the nucleus and cytoplasm of vessel endothelium, gland epithelial, and decidualized stromal cells. In conclusion, FLRG is expressed by the human endometrium, and the different cellular localization suggests novel putative functions. PMID- 15130518 TI - GH suppresses TGF-beta-mediated fibrosis and retains cardiac diastolic function. AB - The aims of this study were to elucidate the molecular mechanism by which growth hormone (GH) excess is anti-fibrotic in vitro and in vivo model. The in vivo model GH excess showed a significant increase of relative wall thickness with no concomitant disturbance of cardiac diastolic function. Western blot for extracellular matrix (ECM) structural proteins showed minimal change in the GH treatment group, compared to an Angiotensin II (Ang II) subpressor dose group. In cultured cardiac fibroblasts, we investigated the abundance of ECM proteins, phosphorylation of p38 mitogen-activated protein kinase (MAPK), and transforming growth factor-beta (TGF-beta)-specific transcriptional activity. GH down regulated the expression of PAI-1 and fibronectin proteins activated by TGF-beta. In reporter assays, GH, but not insulin-like growth factor-1 (IGF-1), reduced TGF beta-specific transcriptional activity. Moreover, GH markedly down-regulated TGF beta-induced phosphorylation of p38 MAPK. These results demonstrated that a chronic excess of GH have an anti-fibrotic effect on cardiac remodeling, probably through a down-regulation of TGF-beta signaling via de-phosphorylation of p38 MAPK. PMID- 15130519 TI - Systemic distribution of estrogen-responsive finger protein (Efp) in human tissues. AB - Estrogen-responsive finger protein (Efp), a target gene product of estrogen receptor (ER), is considered essential for estrogen-dependent cell proliferation. The biological significance of Efp remains unclear in human tissues, and therefore, we examined systemic distribution of Efp in human adult and fetal tissues using RT-PCR and immunohistochemistry. Efp mRNA expression was marked in the placenta and uterus, high in the thyroid gland, aorta, and spleen in adult, and relatively low in other human adult and fetal tissues examined in this study. Efp immunoreactivity was detected in epithelium of various adult tissues, and was also detected in cytotrophoblasts of the placenta and splenic macrophages. Efp immunolocalization in human fetus was generally similar as that in adult. These Efp-positive cells were previously reported to be associated with ERalpha and/or ERbeta expression. Therefore, these results indicate that Efp is widely expressed and may play important roles in various human tissues possibly through ERs. PMID- 15130520 TI - Roles of gonadotropins and meiosis-activating sterols in meiotic resumption of cultured follicle-enclosed mouse oocytes. AB - This study describes a model for short-term culture of intact mouse follicles under serum-free conditions. Follicles were either obtained from immature mice receiving no ovarian stimulation (i.e. no eCG-primed protocol, group I) or from mice undergoing ovarian stimulation (i.e. eCG-primed protocol, group II). Follicles were grouped according to size (100-170, 180-200, 210-250, 260-350 and 360-400 microm, respectively) and cultured for 24h (group I) or for only 6h (group II). Induced meiotic resumption of follicle-enclosed oocytes were evaluated following stimulation with gonadotropins (i.e. FSH and hCG), AY9944-A 7, an inhibitor of Delta14-reductase, and RS-21745, an inhibitor of lanosterol 14alpha-demethylase; both enzymes affect synthesis of the meiosis activating sterols (MAS) that induce oocyte maturation. The frequency of oocyte degeneration was also recorded. In group I, FSH (10-200 IUl-1) and AY9944-A-7 (5, 25 and 50 microM) separately induced resumption of meiosis in oocytes derived from follicles with a diameter of 180-400 microm. hCG (1.0 and 10 IUml-1) exhibited a similar but weaker effect on oocytes present in follicles with a diameter of 260 400 microm. Irrespective of follicular diameter oocytes obtained from follicles in group II responded to hCG and FSH by resuming meiosis. FSH (50 IUl-1) alone or hCG (10 IUml-1) alone both increased the GVBD percentage of oocytes enclosed in follicles with a diameter 260-400 microm, but the response to hCG was not significant compared to control. FSH (50 IUl-1) combination with hCG (10 IUml-1) showed an additive effect raising the rate of GVBD after 6h culture. Addition of 50 or 100 microM RS-21745 was able to attenuate gonadotropins-induced resumption of meiosis to below background levels. In conclusion, the ability of FSH to induce meiotic resumption of follicle-enclosed mouse oocytes is correlated to follicle size, being most pronounced in larger follicles. hCG caused a similar but less pronounced effect. The ability of RS-21745 to inhibit and the ability of AY9944-A-7 to enhance oocyte maturation of follicle-enclosed oocytes support the concept of FSH employing MAS as a downstream signal transduction molecule for initiation of oocyte maturation in mice. PMID- 15130521 TI - Survivin expression in rat testis is upregulated by stem-cell factor. AB - The inhibitor of apoptosis protein BIRC-5/survivin plays roles in both apoptosis and the regulation of chromosome-segregation/cytokinesis during mitosis. As the population dynamics of male germ cells are regulated by both proliferation (mitosis and meiosis) and apoptotic culling, we hypothesized that BIRC-5/survivin could be central to the regulation of spermatogenesis. We have analyzed BIRC 5/survivin expression throughout the seminiferous epithelial cycle of the rat. BIRC-5/survivin RNA and protein exhibit rhythms of expression throughout the seminiferous epithelial cycle. The highest levels of expression were found, by immunohistochemistry and in situ hybridization, to occur during the long first meiotic prophase of spermatocytes. Cytoplasmic abundance declined at metaphase and reappeared at anaphase. Some BIRC-5/survivin expression was also found to occur in interstitial Leydig cells. BIRC-5/survivin protein levels were up regulated in vitro by the paracrine, Stem-Cell Factor, that is known to regulate both proliferation and apoptosis of germ cells and Leydig cells. PMID- 15130522 TI - Ceramide inhibits L-type calcium channel currents in GH3 cells. AB - In this study, we investigated the effect of ceramide on the L-type Ca2+ channel (L-channel) in GH3 cells. We found that C6-ceramide, but not C6-dihydroceramide, the inactive analogue, had an inhibitory effect on BayK 8644-stimulated GH release. Using patch clamp analysis, C6- and C2-ceramide, but not C6 dihydroceramide, were found to inhibit the L-channel current. Increasing intracellular ceramide level with sphingomyelinase also inhibited the L-channel current. The inhibitory effect of ceramide on the L-channel current was attenuated by calphostin C, a myristolated pseudosubstrate protein kinase C (PKC) inhibitor, and lavendustin A, a tyrosine kinase inhibitor. Combined treatment with lavendustin A and the myristolated PKC inhibitor blocked the effect of ceramide on the L-channel current. These results indicate that ceramide, a lipid messenger of the sphingomyelin pathway, is an important regulator of the L channel in GH3 cells and both tyrosine kinase and PKC are involved in this effect of ceramide. PMID- 15130523 TI - Frontal EEG asymmetry, emotion, and psychopathology: the first, and the next 25 years. PMID- 15130524 TI - Frontal EEG asymmetry as a moderator and mediator of emotion. AB - Frontal EEG asymmetry appears to serve as (1) an individual difference variable related to emotional responding and emotional disorders, and (2) a state dependent concomitant of emotional responding. Such findings, highlighted in this review, suggest that frontal EEG asymmetry may serve as both a moderator and a mediator of emotion- and motivation-related constructs. Unequivocal evidence supporting frontal EEG asymmetry as a moderator and/or mediator of emotion is lacking, as insufficient attention has been given to analyzing the frontal EEG asymmetries in terms of moderators and mediators. The present report reviews the frontal EEG asymmetry literature from the framework of moderators and mediators, and overviews data analytic strategies that would support claims of moderation and mediation. PMID- 15130525 TI - Contributions from research on anger and cognitive dissonance to understanding the motivational functions of asymmetrical frontal brain activity. AB - Research has suggested that approach-related positive emotions are associated with greater left frontal brain activity and that withdrawal-related negative emotions are associated with greater right frontal brain activity. Different explanations have been proposed. One posits that frontal asymmetry is due to emotional valence (positivity/negativity), one posits that frontal asymmetry is due to motivational direction (approach/withdrawal), and one posits that frontal asymmetry is due to a combination of emotional valence and motivational direction (positive-approach/negative-withdrawal). Because research had confounded emotional valence and motivational direction, the theoretical explanation was muddled. Solely supporting the motivational direction model, recent research has revealed that anger and cognitive dissonance, emotions with negative valence and approach motivational tendencies, are related to relatively greater left frontal activity. PMID- 15130526 TI - Resting frontal brain activity: linkages to maternal depression and socio economic status among adolescents. AB - We tested the prediction that resting frontal brain asymmetry would be a marker of vulnerability for depression among adolescents. Baseline electroencephalographic (EEG) activity was recorded from 12 to 14-year-old adolescents whose mothers had a history of depression (high risk group) and whose mothers were lifetime-free of axis I psychopathology (low risk group). High risk adolescents demonstrated the hypothesized pattern of relative left frontal hypo activity on alpha-band measures. Such effects were specific to the mid-frontal region and generally consistent across reference montages. Socio-economic status (SES) also predicted alpha asymmetry. When the effects of SES and risk status were jointly assessed, SES contributed unique variance to the prediction of frontal brain asymmetry. The implications of the observed relations among maternal depression, SES, and frontal brain asymmetry are discussed. PMID- 15130527 TI - Patterns of brain electrical activity in infants of depressed mothers who breastfeed and bottle feed: the mediating role of infant temperament. AB - Successful breastfeeding involves a dyadic interaction between a mother and her infant. The present study was designed to examine the association between breastfeeding and temperament in infants of depressed mothers. Seventy-eight mothers, 31 who were depressed, and their infants participated. Depressed mothers who had stable breastfeeding patterns were less likely to have infants with highly reactive temperaments. Multivariate analyses of variances (MANOVAs) showed that infants of depressed mothers who breastfed did not show the frontal asymmetry patterns, i.e., left frontal hypoactivity, previously reported. Moreover, breastfeeding stability, even in depressed mothers, was related to more positive dyadic interactions. Finally, a model was supported, in which the effects of maternal depression on infant feeding are mediated by infant frontal EEG asymmetry and infant temperament. These findings could provide a foundation for developing intervention techniques, employing breastfeeding promotion and support, directed toward attenuating the affective and physiological dysregulation already noted in infants of depressed mothers. PMID- 15130528 TI - Prefrontal cortex activity differentiates processes affecting memory in depression. AB - Deficits in the initiation and utilization of strategies contribute importantly to memory impairments in depression. Other research on depression has documented memory biases toward negative and away from positive material. This study investigated brain mechanisms accompanying the initiative deficit and negative bias processes affecting memory in depressed individuals. Electroencephalography was recorded prior to and during emotional narratives and correlated with subsequent memory recognition of narrative material. Hypothesized to reflect strategy initiation, bilateral activity of the prefrontal cortex (PFC) preceding a sad narrative was associated with memory performance for that narrative in nondepressed controls only. Negative memory bias in depressed participants was inferred from their association between right prefrontal activity during the sad narrative and memory performance, consistent with research implicating that region in withdrawal-related unpleasant emotions. These results highlight the importance of distinguishing processes that influence memory performance when investigating the neural mechanisms of cognitive deficit and bias in depression. PMID- 15130529 TI - Relative left-frontal activity is associated with increased depression in high reassurance-seekers. AB - Excessive reassurance-seeking, which has been associated with depression in many studies, can be defined as the relatively stable tendency to seek assurance perseveratively from others. We hypothesized that although depression has been associated with left-frontal EEG hypoactivity, reassurance-seekers may possess a unique diathesis that is more likely to be associated with increased left-frontal activity. Data were collected from 12 volunteers who were receiving therapeutic services from a University Clinic. EEG asymmetry scores were averaged over two measurement occasions at least 3 weeks apart. As predicted, stable relative right frontal activity was associated with increased depression in those who were low on reassurance-seeking, while stable relative left-frontal activity was associated with increased depression among high reassurance-seekers. Perhaps those who seek reassurance excessively do so because of their inability to alter their behavior even when environmental cues are no longer reinforcing, which can maintain or exacerbate their depressive symptoms. PMID- 15130530 TI - Individual differences in anterior EEG asymmetry: methodological problems and solutions. AB - In the last two decades, a substantive body of empirical work investigated the association between individual differences of electroencephalogram (EEG) alpha asymmetry and affective/motivational dispositions. Recent work indicated that several methodological problems persist in this field. The present paper reviews these problems with a focus on the reliability and validity of measures of anterior resting EEG asymmetry, which serve as a proxy for trait-like asymmetries of cortical activity. These issues include the treatment of ocular and muscle artifacts, the choice of the EEG reference, the use of current source density (CSD) measures, the state-trait nature of resting asymmetry, and the treatment of state-like fluctuations of the measures. In addition, the statistical problem of inflated type-I error due to multiple testing is also considered and different approaches to counteract this problem are suggested. PMID- 15130531 TI - Issues and assumptions on the road from raw signals to metrics of frontal EEG asymmetry in emotion. AB - There exists a substantial literature examining frontal electroencephalographic asymmetries in emotion, motivation, and psychopathology. Research in this area uses a specialized set of approaches for reducing raw EEG signals to metrics that provide the basis for making inferences about the role of frontal brain activity in emotion. The present review details some of the common data processing routines used in this field of research, with a focus on statistical and methodological issues that have captured, and should capture, the attention of researchers in this field. PMID- 15130532 TI - What does the prefrontal cortex "do" in affect: perspectives on frontal EEG asymmetry research. AB - This commentary provides reflections on the current state of affairs in research on EEG frontal asymmetries associated with affect. Although considerable progress has occurred since the first report on this topic 25 years ago, research on frontal EEG asymmetries associated with affect has largely evolved in the absence of any serious connection with neuroscience research on the structure and function of the primate prefrontal cortex (PFC). Such integration is important as this work progresses since the neuroscience literature can help to understand what the prefrontal cortex is "doing" in affective processing. Data from the neuroscience literature on the heterogeneity of different sectors of the PFC are introduced and more specific hypotheses are offered about what different sectors of the PFC might be doing in affect. A number of methodological issues associated with EEG measures of functional prefrontal asymmetries are also considered. PMID- 15130533 TI - Feelings and emotions: roles for electrophysiological markers. AB - Asymmetrical electroencephalogram (EEG) alpha activity over anterior regions of the scalp predicts a variety of outcome measures of interest to emotion researchers. This vast and diverse literature is examined from three different viewpoints. First, the organization of this vast literature is contrasted from theoretical and statistical perspectives, and the advantages and disadvantages of each perspective are considered. Second, the correlates of EEG asymmetry are sometimes treated as criterion (dependent) measures and at other times treated as predictor (independent) measures. Differences in the interpretation of each are surveyed, and the need for attention to whether EEG asymmetry is a simple correlate, mediator, or moderator of the associated affective measures is noted. Finally, the studies of EEG asymmetry and emotion that adopt a psychological perspective are contrasted with those that adopt a neurophysiological perspective, and the import of each for theory, experimental design, and analytic strategy is discussed. PMID- 15130534 TI - In vivo examination of hydroxyurea and the novel ribonucleotide reductase inhibitors trimidox and didox in combination with the reverse transcriptase inhibitor abacavir: suppression of retrovirus-induced immunodeficiency disease. AB - Inhibition of ribonucleotide reductase (RR) has gained attention as a potential strategy for HIV-1 therapy through the success of hydroxyurea (HU) to potentiate the activity of the nucleoside reverse transcriptase inhibitor (NRTI) didanosine (ddI) in clinical trials. However, the use of HU has been limited by its development of hematopoietic toxicity. In this study, the novel RR inhibitors didox (DX; 3,4-dihydroxybenzohydroxamic acid), and trimidox (TX; 3,4,5 trihydroxybenzamidoxime) were evaluated along with HU for anti-retroviral efficacy in LPBM5-induced retro-viral disease (MAIDS) both as monotherapeutic regimens and in combination with the guanine containing NRTI abacavir (ABC). Anti retroviral drug efficacy was determined by measuring inhibition of splenomegaly, hypergammaglobulinemia, and splenic levels of proviral DNA. In this study, all RRIs tested showed the ability to improve the efficacy of ABC in the MAIDS model by reducing splenomegaly, hypergammaglobulinemia, and splenic proviral DNA levels. PMID- 15130535 TI - Evaluation of antiviral activity against human herpesvirus 8 (HHV-8) and Epstein Barr virus (EBV) by a quantitative real-time PCR assay. AB - A real-time quantitative PCR was developed to assess antiviral activity of molecules against human herpesvirus 8 (HHV-8) and the Epstein-Barr virus (EBV). The antiviral activity of the reference molecules acyclovir, ganciclovir, cidofovir, adefovir and brivudin, as assessed by this methodology, proved very similar to the activity as determined by a DNA-DNA hybridisation method. PMID- 15130536 TI - Structural and functional features of the polycationic peptide required for inhibition of herpes simplex virus invasion of cells. AB - Glycoprotein C (gC) of herpes simplex virus type 1 (HSV-1) mediates initial virus contact with cells by binding to heparan sulfate (HS) chains. The synthetic peptide 137GSRVQIRCRFRNSTR151 overlapping a major part of the HS-binding site of gC inhibited HSV-1 infection and, to some extent, HSV-2 infection of cells. Experiments on mutant, glycosaminoglycan-deficient cells as well as the binding assays involving peptide and purified cell surface components identified HS, and, to a lesser degree, chondroitin sulfate as sites of peptide activity. Anti-HSV-1 activity of the peptide was due to (i) partial inhibition of virus binding to cells and (ii) arresting the virions, which managed to attach to the cells in the presence of peptide, at a step of initial relatively weak binding. Analysis of the ionic-strength dependence of the peptide-HS and the virus-HS interactions revealed that the more efficient inhibition by the peptide of HSV-1 than HSV-2 infectivity was due to a relatively high affinity of HSV-2 for HS, a feature of importance in overcoming the peptide block. Mutational analysis of viral gC and peptide variants identified, apart from basic amino acids, two hydrophobic residues Ile(142) and Phe(146) as important in maintaining the specific affinity of peptide for HS and, hence, its anti-HSV activity. These results could contribute to the development of anti-HSV compounds that target initial events in the virus-cell interaction. PMID- 15130537 TI - Improved antiviral activity in vitro of ribavirin against measles virus after complexation with cyclodextrins. AB - Despite vaccination, measles remains a burden in both developed and developing countries and complications may necessitate an efficient therapy. Measles virus (MEV) is susceptible to ribavirin (RBV), but the use of this drug is limited by its toxicity. Cyclodextrins (CDs) can form complexes with numerous molecules, improving their bioavailability and their biological properties. We have evaluated in vitro the antiviral effects of complexes of RBV with alpha-, beta- or gamma-CD against two clade A laboratory strains of MEV (Edmonston and CAM/RB) grown on Vero cells. Complexation of RBV with alpha-CD or beta-CD lead to a five fold or a two-fold decrease in the 50% inhibitory concentration, respectively, against both MEV strains. In contrast, gamma-CD complexation showed no modification. PMID- 15130538 TI - Identification of related gene/protein names based on an HMM of name variations. AB - Gene and protein names follow few, if any, true naming conventions and are subject to great variation in different occurrences of the same name. This gives rise to two important problems in natural language processing. First, can one locate the names of genes or proteins in free text, and second, can one determine when two names denote the same gene or protein? The first of these problems is a special case of the problem of named entity recognition, while the second is a special case of the problem of automatic term recognition (ATR). We study the second problem, that of gene or protein name variation. Here we describe a system which, given a query gene or protein name, identifies related gene or protein names in a large list. The system is based on a dynamic programming algorithm for sequence alignment in which the mutation matrix is allowed to vary under the control of a fully trainable hidden Markov model. PMID- 15130539 TI - The difficult interpretation of transcriptome data: the case of the GATC regulatory network. AB - Genomic analyses on part of Escherichia coli's chromosome had suggested the existence of a GATC regulated network. This has recently been confirmed through a transcriptome analysis. Two hypotheses about the molecular control mechanism have been proposed-(i) the GATC network regulation is caused by the presence of GATC clusters within the coding sequences; the regulation is the direct consequence of the clusters' hemi-methylation and therefore their elevated melting temperature, (ii) the regulation is caused by the presence of GATCs in the non-coding 500 bp upstream regions of the affected genes; it is the consequence of an interaction with a regulatory protein like Fnr or CAP. An analysis of the transcriptome data has not allowed us to decide between the two hypotheses. We have therefore taken a classic genomic approach, analyzing the statistical distribution of GATC along the chromosome, using a realistic model of the chromosome as theoretical reference. We observe no particular distribution of GATC in the non-coding upstream regions; however, we confirm the presence of GATC clusters within the genes. In order to verify that the particular distribution observed in E. coli is not a statistical artefact, but has a physiological role, we have carried out the same analysis on Salmonella, making the hypothesis that the genes containing a GATC clusters should be largely the same in the two bacteria. This has been indeed observed, showing that the genes containing a GATC cluster are part of a regulation network. The present is a case study, which demonstrates that the analysis of transcriptome data does not always permit to identify the primary cause of a phenomenon observed; on the other hand, a classic genomic approach linked with a comparative study of related genomes may allow this identification. PMID- 15130540 TI - MoDEL: an efficient strategy for ungapped local multiple alignment. AB - We introduce a method for ungapped local multiple alignment (ULMA) in a given set of amino acid or nucleotide sequences. This method explores two search spaces using a linked optimization strategy. The first search space M consists of all possible words of a given length W, defined on the residue alphabet. An evolutionary algorithm searches this space globally. The second search space P consists of all possible ULMAs in the sequence set, each ULMA being represented by a position vector defining exactly one subsequence of length W per sequence. This search space is sampled with hill-climbing processes. The search of both spaces are coupled by projecting high scoring results from the global evolutionary search of M onto P. The hill-climbing processes then refine the optimization by local search, using the relative entropy between the ULMA and background residue frequencies as an objective function. We demonstrate some advantages of our strategy by analyzing difficult natural amino acid sequences and artificial datasets. A web interface is available at PMID- 15130541 TI - A note on clustering the functionally-related paralogues and orthologues of proteins: a case of the FK506-binding proteins (FKBPs). AB - The expression patterns of 18 FK506-binding proteins (FKBPs) encoded in the human genome have been established whereas the functional significance of the numerous ORFs coding for FKBP-like sequences remains unknown. Nominal masses of the human FKBPs vary from 12 to 135 kDa. Some large FKBPs consist up to four repeats of the 12 kDa FK506-like binding domain (FKBD) whereas other large FKBPs contain one FKBD linked to different functional domains such as TPRs, leucine-zipper, calmodulin-binding domain etc. The genomes of other eukaryotic organisms, namely D. melanogaster, C. elegans, A. thaliana, S. pombe and S. cerevisiae encode different numbers of the FKBPs' paralogues some of which are orthologues to the human FKBPs. A library of novel algorithms was developed and used for computation of the level of conservation of the hydrophobicity and bulkiness profiles, and the amino acid compositions (AACs) of 247 aligned sequences of FKBPs. The pairwisely-compared hydrophobicity and bulkiness profiles for some combinations of the aligned sequences of the FKBDs yielded high values of the correlation coefficients (CCF). The AACs of some combinations of the aligned sequences of the FKBDs also differed to a low degree. The functionally-related orthologues and paralogues of the FKBPs were clustered by using the following criteria: 1 degrees apparent conservation of the crucial amino acid (AA) residues for peptidylprolyl cis/trans isomerase (PPIase) acitity and binding of some immunosuppressive drugs; 2 degrees convergence of the three mentioned above properties of the polypeptide chain; 3 degrees similarity in the sequence attributes pI and total hydrophobicity index (HI). The clustering method was used for setting up several hypotheses on the emergence of certain classes of the FKBPs in the eukaryotic kingdom. PMID- 15130542 TI - An adaptive and iterative algorithm for refining multiple sequence alignment. AB - Multiple sequence alignment is a basic tool in computational genomics. The art of multiple sequence alignment is about placing gaps. This paper presents a heuristic algorithm that improves multiple protein sequences alignment iteratively. A consistency-based objective function is used to evaluate the candidate moves. During the iterative optimization, well-aligned regions can be detected and kept intact. Columns of gaps will be inserted to assist the algorithm to escape from local optimal alignments. The algorithm has been evaluated using the BAliBASE benchmark alignment database. Results show that the performance of the algorithm does not depend on initial or seed alignments much. Given a perfect consistency library, the algorithm is able to produce alignments that are close to the global optimum. We demonstrate that the algorithm is able to refine alignments produced by other software, including ClustalW, SAGA and T COFFEE. The program is available upon request. PMID- 15130543 TI - A nucleotide composition constraint of genome sequences. AB - Let a, c, g and t denote the occurrence frequencies of A, C, G and T, respectively, in a genome. We calculated the statistical quantity S = a2 + c2 + g2 + t2 for each of 809 genomes (11 archaea, 42 bacteria, 3 eukaryota, 90 phages, 36 viroids and 627 viruses) and 236 plasmids. We found that S < 1/3 is strictly valid for almost all of the above genomes or plasmids. As a direct deduction of the above observation, it is shown that (i) the statistical quantity S is a kind of genome order index, which is negatively correlated with the Shannon H function; (ii) S < 1/3 suggests that a minimal value of the Shannon H function is required for each genome; (iii) S defined above would be a new biological statistical quantity, useful to describe the composition features of genomes; (iv) By jointly considering the Chargaff Parity Rule 2, it is shown that the genomic G + C content should be in between 0.211 and 0.789. PMID- 15130544 TI - Matrix, reinvention in plants: how genetics is unveiling secrets of non-host disease resistance. PMID- 15130545 TI - Clustering of centromeres precedes bivalent chromosome pairing of polyploid wheats. PMID- 15130546 TI - New signalling molecules regulating root hair tip growth. PMID- 15130547 TI - Bifunctional and moonlighting enzymes: lighting the way to regulatory control. PMID- 15130548 TI - Phospholipid-derived signaling mediated by phospholipase A in plants. PMID- 15130549 TI - Plant PP2C phosphatases: emerging functions in stress signaling. PMID- 15130550 TI - Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. PMID- 15130551 TI - Interactions of insect pheromones and plant semiochemicals. PMID- 15130552 TI - Do robots need to sleep? AB - The present paper exposes algorithmic results providing a vision about sleep functions which complements biological theory and experiments. Derived from the algorithmic theory of information, the theory of adaptation aims at quantifying how an inherited or acquired piece of knowledge helps individuals to survive. It gives a scale of complexity for survival problems and proves that some of them can only be solved by a dynamical management of memory associating continuous learning and forgetting methods. In this paper we explain how a virtual robot "Picota" has been designed to simulate the behavior of a living hen. In order to survive in its synthetical environment, our robot must recognize good seeds from bad ones, and should take rest during night periods. Within this frame, and facing the rapid evolution of to-be-recognized forms, the best way to equilibrate the energetic needs of the robot and ensure survival is to use the nightly rest to reorganize the pieces of data acquired during the daily learning, and to trash the less useful ones. Thanks to this time sharing, the same circuits can be used for both daily learning and nightly forgetting and thus costs are lower; however, this also forces the system to "paralyse" the virtual robot, and therefore the night algorithm is reminiscent of paradoxical (REM) sleep. The algorithm of the robot takes advantage of the alternation between wakefulness or activity and the rest period. This diagram quite accurately recalls the REM period. In the future, the convergence between the neurophysiology of sleep and the theory of complexity may give us a new line of research in order to elucidate sleep functions. PMID- 15130553 TI - Atypical electrophysiologic findings in chronic inflammatory demyelinating polyneuropathy (CIDP)--diagnosis confirmed by nerve biopsy. AB - OBJECTIVES: Numerous sets of electrophysiological criteria of chronic inflammatory demyelinating polyneuropathy (CIDP) have been proposed, among which the criteria established by an ad hoc subcommittee of the American Academy of Neurology (AAN) in 1991 (Neurology 41 (1991) 617) are the most widely used. As they seemed rather restrictive, the Inflammatory Neuropathy Cause and Treatment (INCAT) group (Ann. Neurol. 50 (2001) 195) proposed modifications of these electrophysiological criteria. However, even using these criteria, some cases of CIDP may not be recognized. In such cases, nerve biopsy has proven useful for confirmation of the diagnosis by demonstrating specific abnormalities. The objective of the study was to determine the profile of electrophysiological abnormalities in patients with atypical electrophysiologic criteria of CIDP and the diagnostic value of multiple A waves and a low median to sural amplitude ratio. PATIENTS AND METHODS: Over a period of 3 years, we classified 44 patients into two categories: those presenting the strict AAN and/or INCAT criteria and those who we regarded as cases of CIDP not meeting these criteria. All patients benefited from one or more clinical and electrophysiological examination. Extensive biological workup and genetic study when appropriate excluded other causes of neuropathy. Nerve biopsies were taken from all patients and samples were included in paraffin and epon for systematic light, teasing and electron microscopic examination. RESULTS AND CONCLUSION: Out of 44 patients, 36 fulfilled the INCAT or AAN criteria. In eight other patients, the diagnosis of CIDP was suspected on clinical and EMG examinations and confirmed by nerve biopsy. In these cases, the electrophysiological exploration showed some abnormalities such as multiple A waves in four out of eight patients or an abnormal pattern of the sensory responses of the median and sural nerves in four out of eight patients that were more indicative of an initial demyelinating process. Six of our patients received immunomodulatory treatment, and five responded favorably. PMID- 15130555 TI - Neuropathic pain controlled for more than a year by monthly sessions of repetitive transcranial magnetic stimulation of the motor cortex. AB - Neuropathic pain can be controlled by motor cortex stimulation using surgically implanted electrodes in a majority of selected patients. Analgesic effects were also found to result from repetitive transcranial magnetic stimulation (rTMS) of the cortex. We report the case of a woman, in whom drug-resistant peripheral pain was controlled for 16 months by monthly sessions of motor cortex rTMS until a durable pain relief was obtained after surgical implantation of a cortical stimulator. This case illustrates the value of rTMS in helping patients to wait for surgery. PMID- 15130554 TI - Use of gabapentin for attenuation of symptoms following rapid opiate detoxification (ROD)--correlation with neurophysiological parameters--. AB - Rapid opiate detoxification (ROD) is a technique whereby the opiate-dependent patient is withdrawn acutely, under anesthesia, from the opioid. Following detoxification, patients experience severe back pain and restlessness often accompanied by a restless-leg-syndrome. We evaluated gabapentin given immediately following detoxification to attenuate these symptoms. In addition, we evaluated the use of the somatosensory-evoked potential (SEP) as a parameter to quantitate pain responses. Patients (n=21; mean age 32.5 +/- 7 SD; 12 males, 9 females) underwent ROD with naltrexone (2 x 50 mg) during propofol anesthesia and artificial ventilation (IPPV). Sympathetic overshoot was attenuated by clonidine, and increased bowl movement was managed by continuous i.v. somatostatin. Back pain, restlessness, and restless-leg-syndrome were treated with gabapentin (1200 mg) in the ICU. Efficacy was assessed by the patient's subjective ratings of restlessness (0-4). In addition, measurements of amplitude (microV), latency (ms) of late N100-peak of the somatosensory evoked potential (SEP), and tolerance to an increased electrical nociceptive stimulus (mA) to the forearm were performed. Data were compared to pre-treatment control and to the period shortly after detoxification. From a mean of 8.4 +/- 2.5 microV, N100-peak increased to a mean of 12.3 microV +/- 3.3 (p < 0.005) following opioid detoxification. Gabapentin reduced amplitude height to a mean of 3.5 +/- 1.5 microV. Also, tolerance to nociceptive stimulus, which had dropped to 4.4 mA, increased to 12.5 mA (p < 0.01), while intensity for restlessness and thrashing of limbs dropped from 3.2 to 1.2 (p < 0.05). The sudden displacement of the opiate from its receptor site induced by naltrexone, resulted in a post inhibitory SEP overshoot with an increase in nociceptive afferent volleys, and a lowering in pain threshold. This was associated with back pain, limb thrashing and a restless-leg-syndrome, all of which could be attenuated by gabapentin. The amplitude of late N100-peak parameter appears to be a potential candidate to quantify the increase of nociception in such patients. PMID- 15130556 TI - Break on through to the other side--the Sec translocon. PMID- 15130557 TI - A novel conserved family of nitric oxide synthase? PMID- 15130558 TI - Identifying protein kinase substrates: hunting for the organ-grinder's monkeys. PMID- 15130559 TI - Advances in protein kinase B signalling: AKTion on multiple fronts. PMID- 15130560 TI - S-adenosylmethionine: nothing goes to waste. AB - S-adenosylmethionine (SAM or AdoMet) is a biological sulfonium compound known as the major biological methyl donor in reactions catalyzed by methyltransferases. SAM is also used as a source of methylene groups (in the synthesis of cyclopropyl fatty acids), amino groups (in the synthesis of 7,8-diaminoperlagonic acid, a precursor of biotin), ribosyl groups (in the synthesis of epoxyqueuosine, a modified nucleoside in tRNAs) and aminopropyl groups (in the synthesis of ethylene and polyamines). Even though the mechanism of most of these reactions has not been extensively characterized, it is likely that the chemistry at work is mainly driven by the electrophilic character of the carbon centers that are adjacent to the positively charged sulfur atom of SAM. In addition, SAM, upon one electron reduction, is a source of 5'-deoxyadenosyl radicals, which initiate many metabolic reactions and biosynthetic pathways by hydrogen-atom abstraction. SAM presents a unique situation in which all constituent parts have a chemical use. PMID- 15130561 TI - Relationship of two human tRNA synthetases used in cell signaling. AB - Human tyrosyl-tRNA synthetase (TyrRS) and tryptophanyl-tRNA synthetase (TrpRS) are closely related, dual function enzymes that act in protein biosynthesis and angiogenesis. The recent crystallographic structures of these two enzymes show that they adopt remarkably similar three-dimensional (3D) architectures, being more like each other than like their respective prokaryotic orthologs. In particular, adaptations to the anticodon recognition domain of TyrRS cause distinct appended domains in TyrRS and TrpRS to occupy the same 3D space and thus to mask a common surface on each synthetase. Thought to be important for cell signaling activity, this surface is made accessible by proteolytic cleavage, thereby activating the cell-signaling function of these enzymes. PMID- 15130562 TI - Structural aspects of sexual attraction and chemical communication in insects. AB - In the animal kingdom, the relationship between individuals and the environment is often modulated by chemical communication. In the olfaction of insects, small antennary proteins--such as odorant- and pheromone-binding proteins (OBPs and PBPs, respectively) and chemosensory proteins (CSP)--have been proposed to carry their ligand from the air-fluid interface to the olfactory receptors. Binding experiments and recent structural studies of three PBPs, an OBP and a CSP have illustrated their versatility and ability to accommodate ligands of different shapes and chemical structures. The role of these proteins--as simple transporters or receptor triggers--is still a matter of debate, but some recent data seem to support the latter. PMID- 15130563 TI - New insights into TGF-beta-Smad signalling. AB - Transforming growth factor beta (TGF-beta) initiates its diverse cellular responses by binding to and activating specific cell surface receptors that have intrinsic serine/threonine kinase activity. These activated TGF-beta receptors stimulate the phosphorylation of receptor-regulated Smad proteins, which in turn form complexes with Smad4 that accumulate in the nucleus and regulate the transcription of target genes. TGF-beta responses can be cell-type specific and are dependent on both the concentration of TGF-beta signalling components and the activity of other signal transduction pathways, which can either synergize with or antagonize the TGF-beta pathway. Recent research has provided insights into the specificity determinants of TGF-beta-Smad signalling, including combinatorial ligand-receptor associations, selective interactions between the Smads and other pathway components that are mediated through defined binding motifs, and the differential regulation of duration and intensity of signalling. PMID- 15130564 TI - The antibody-catalyzed water oxidation pathway--a new chemical arm to immune defense? AB - Antibodies are the classical adaptor molecules of the immune system, linking recognition and killing of foreign pathogens. However, the recent discovery of a new property of the antibody molecule suggests a previously unexplored effector function of the immune system. All immunoglobulins, regardless of source or antigenic specificity, can catalyze the reaction between singlet (1Deltag) oxygen and water to give hydrogen peroxide (H(2)O(2)). Both the chemical and biological aspects of this pathway are being explored and intriguing new insights into how this pathway might have a role in immune defense are emerging. PMID- 15130566 TI - Can practitioners be researchers? PMID- 15130567 TI - Providing Complementary and Alternative Medicine in primary care: the primary care workers' perspective. AB - BACKGROUND: The use of Complementary and Alternative Medicine (CAM) in primary care is growing, but still not widespread. Little is known about how CAM can/should be integrated into mainstream care. OBJECTIVES: To assess primary care health professionals' perceptions of need and of some ways to integrate CAM in primary care. METHOD: Questionnaire survey of primary health care workers in Northwest London. General Practitioners (GPs) were targeted in a postal survey, other members of the primary care team, such as district and practice nurses, were targeted via colleagues. The questionnaire assessed health care professionals' perspective on complementary medicine, referrals, ways to integrate complementary medicine into primary care and interest in research on CAM. RESULTS: Responses were obtained from 149 GPs (40% response rate after one reminder) and 24 nurses and 32 other primary care team members. One hundred and seventy-one (83%) respondents had previously referred (or influenced referral) for CAM treatments, the main reasons cited were: patients request (68%), conventional treatments failed (58%) and evidence (36%) (more than one reason could be given). Acupuncture and homoeopathy were the therapies for which patients were most frequently referred, followed by manual therapies. There was a significant interest in more training/information on CAM (66%). Only 12 respondents (6%) were against any integration of CAM in mainstream primary care. Most respondents felt that CAM therapies should be provided by doctors (66%) or other health professionals trained in CAM (82%). Twenty-six percent of respondents agreed with provision of CAM by non-state-registered practitioners. It was felt that the integration of CAM could lead to cost savings (70%), particularly in conditions involving pain, but also cost increases (55%) particularly in 'poorly defined conditions'. Fifty-six percent of respondents would consider participating in studies investigating CAM. The greatest interest was in acupuncture (41% of those who expressed an interest in research), homoeopathy (30%) and therapeutic massage/aromatherapy (26%). CONCLUSIONS: There is considerable interest in CAM among primary care professionals, and many are already referring or suggesting referral. Such referrals are driven mainly by patient demand and by dissatisfaction with the results of conventional medicine. Most of our respondents were in favour of integrating at least some types of CAM in mainstream primary care. There is an urgent need to further educate/inform primary care health professionals about CAM. PMID- 15130568 TI - Naturopathic and Western herbal medicine practice in Australia-a workforce survey. AB - BACKGROUND: Despite substantial growth in the use of complementary medicine, no comprehensive national study has been undertaken of the naturopathic and Western herbal medicine component of the healthcare workforce in Australia. This study aimed to examine the nature of these practices and this currently unregulated workforce in Australia. METHODS: A comprehensive survey questionnaire was developed in consultation with the profession and distributed nationally to all members of the naturopathic and Western herbal medicine workforce. RESULTS: The practices of herbal medicine and naturopathy make up a sizeable component of the Australian healthcare sector, with approximately 1.9 million consultations annually and an estimated turnover of $AUD 85 million in consultations (excluding the cost of medicines). A large proportion of patients are referred to practitioners by word of mouth. Up to one third of practitioners work in multidisciplinary clinics with other registered sectors of the healthcare community. The number of adverse events associated with herbal medicines, nutritional substances and homoeopathic medicines recorded in Australia is substantial and the types of events reported are not trivial. Data suggest that practitioners will experience one adverse event every 11 months of full-time practice, with 2.3 adverse events for every 1000 consultations (excluding mild gastrointestinal effects). CONCLUSION: These data confirm the considerable degree of utilisation of naturopathic and Western herbal medicine practitioners by the Australian public. However, there is a need to examine whether statutory regulation of practitioners of naturopathy and Western herbal medicine is required to better protect the public. PMID- 15130569 TI - The effectiveness of relaxation acupoint stimulation and acupressure with aromatic lavender essential oil for non-specific low back pain in Hong Kong: a randomised controlled trial. AB - OBJECTIVES: To assess the effect of acupoint stimulation with electrodes combined with acupressure using an aromatic essential oil (lavender) as an add-on treatment on pain relief and enhancing the physical functional activities among adults with sub-acute or chronic non-specific low back pain. DESIGN: Randomised controlled trial. SETTING: The community centre, Old-Aged Home and Women Workers Association, Hong Kong. INTERVENTION: 8-session relaxation acupoint stimulation followed by acupressure with lavender oil over a 3-week period. The control group received usual care only. OUTCOME MEASURES: Changes from baseline to the end of treatment were assessed in pain intensity (by Visual Analogue Scale) and duration; lateral fingertip-to-ground distance in centimetres; walking time and interference on daily activities. RESULTS: The baseline VAS scores for the intervention and control groups were 6.38 (S.E.M. = 0.22) and 5.70 (S.E.M. = 0.37) out of 10, respectively ( P=0.24 ). One week after the end of treatment, the intervention group had 39% greater reduction in VAS pain intensity than the control group ( P=0.0001 ), improved walking time ( P=0.05 ) and greater lateral spine flexion range ( P=0.01 ). CONCLUSIONS: Our results show that 8-sessions of acupoint stimulation followed by acupressure with aromatic lavender oil were an effective method for short-term LBP relief. No adverse effects were reported. To complement mainstream medical treatment for sub-acute LBP, the combined therapy of acupoint stimulation followed by acupressure with aromatic lavender oil may be one of the choices as an add-on therapy for short-term reduction of LBP. PMID- 15130570 TI - Acupuncture for low back pain: traditional diagnosis and treatment of 148 patients in a clinical trial. AB - AIMS: To assess patterns of diagnosis, including concordance, and treatment within a clinical trial of traditional acupuncture for low back pain. SETTING: In a pragmatic randomised controlled clinical trial, 148 patients with low back pain, of between 4 weeks and 12 months duration, were randomised to the offer of individualised acupuncture and received up to 10 treatments. METHODS: Standardised diagnosis and treatment records were completed by practitioners for 148 patients. The diagnosis was based on three pre-defined low back pain syndromes. For a subgroup of patients, one of the six practitioners then independently re-examined the patients, blind to the original diagnosis. The diagnostic inter-rater reliability was assessed in terms of percentage congruent classifications and Cohen's Kappa. Structured interviews of practitioners established further details about practice styles. RESULTS: The most commonly diagnosed syndrome associated with low back pain was Qi and Blood Stagnation (88% of patients), followed by Kidney Deficiency (53%) and Bi Syndrome (28%), with more than one syndrome being identified for 65% of patients. For the subgroup examined twice, practitioner concordance was reasonable: between 47 and 80% of classifications were congruent, while Kappa values lay between 0 ("the same as chance") and 0.67 ("good"). Practitioners provided 1269 treatments in total, using 177 different acupuncture points. Most commonly used channels were Bladder and Gall Bladder, and the commonest points were BL-23 and the two lowest Huatuojiaji points. Auxiliary treatments were utilised by all practitioners to varying degrees. CONCLUSIONS: Diagnostic concordance among practitioners was reasonable, and clear themes emerged for treatment. Further research is required to develop a flexible trial protocol with scope for individualised treatment. PMID- 15130571 TI - An alternative treatment for pityriasis versicolor, tinea cruris, tinea corporis and tinea faciei with topical application of honey, olive oil and beeswax mixture: an open pilot study. AB - OBJECTIVE: To evaluate the possible role of honey, olive oil and beeswax in the treatment of skin fungal infections. PATIENTS AND METHODS: Thirty-seven patients with pityriasis versicolor, tinea cruris, tinea corporis and tinea faciei were studied. After clinical evaluation of redness, scaling, pruritus and burning/pain sensation and mycological assessment, honey mixture containing honey, olive oil and beeswax (1:1:1) was applied to the lesions three times daily for a maximum of 4 weeks. RESULTS: Clinical response was obtained in 86% of patients with pityriasis versicolor, 78% of patients with tinea cruris and in 75% of patients with tinea corporis. Mycological cure was obtained in 75, 71 and 62% of patients with PV, tinea cruris and tinea corporis, respectively. The patient with tinea faciei showed clinical and mycological cure 3 weeks after commencement of therapy. CONCLUSION: Honey mixture may have place in the management of these skin conditions and rigorous, controlled trials are justified. PMID- 15130572 TI - A survey of patients' out-of-pocket payments for complementary and alternative medicine therapies. AB - OBJECTIVE: Investigate the out-of-pocket payments for complementary and alternative therapies. DESIGN: Cross-sectional study using the 1996 Medical Expenditure Panel Survey. SETTING: Fourteen thousand and twenty-six US adults aged > or =25 years from 10,597 answering households. Respondents to the survey were only included in this analysis if they used a single CAM therapy which was not covered by insurance. MAIN OUTCOME MEASURE: Reported out-of-pocket payments. RESULTS: Out of 836 users of alternative therapies, 577 used only a single therapy. Massage therapy (23.7%), spiritual healing (16.6%) and herbal therapy (14.7%) were the most frequently single used therapies. The out-of-pocket payments per visit for providers was on average $49 for nutritional advice, $44 for acupuncture, $33 for massage, $23 for herbal therapies, and $9 for spiritual healing. CONCLUSIONS: These are the first estimates of the out-of-pocket payments for specific complementary and alternative therapies. They can help understand the importance people place on these therapies. PMID- 15130573 TI - A survey of attitudes to Traditional Chinese Medicine in Hong Kong pharmacy students. AB - OBJECTIVES: To study the attitudes and personal experiences with Traditional Chinese Medicine (TCM) use in pharmacy students. DESIGN: Prospective cross sectional study. SETTING: University School of Pharmacy. METHODS: Pharmacy students were asked 17 questions according to an anonymous survey questionnaire. RESULTS: All 91 students (46% males) participated in the survey. The attitude toward TCM use was positive in 40%, neutral in 59% and negative in only 1%. On scales from 0 to 10, the mean (SD) scores for the adequacy of the current curriculum in TCM training and state of TCM knowledge were 3.2 (2.0) and 3.6 (1.6), respectively. Of the 35 participants who had used TCM in the past year, nearly half did so without any TCM practitioner consultation. Seventy-four percent reported that they had not been told of any side effects of TCM. When comparing the third year with first or second year students, there was no significant difference between any of these findings. CONCLUSION: The training in TCM within this 3-year pharmacy curriculum appears inadequate. Most students are not aware of any possible side effects in TCM. The attitude and practice do not appear to vary significantly between successive year of pharmacy students. PMID- 15130574 TI - Misuse of a herb: stinging nettle (Urtica urens) induced severe tongue oedema. PMID- 15130575 TI - Mitochondrial building blocks. AB - Despite many genomic and proteomic attempts, approximately half of all mitochondrial proteins remain unidentified. Moreover, the composition of mitochondria varies in different mammalian cell types and the details of this tissue specificity are unclear. Two recent reports provide a major advance in our understanding of mitochondrial function. Sickmann et al. used an exhaustive proteomic approach and came very close to identifying the complete set of yeast mitochondrial proteins. Mootha et al. examined mitochondria from mouse brain, heart, kidney and liver cells, finding that a surprising fraction of the proteins are expressed in only a subset of tissues. PMID- 15130576 TI - O-GlcNAc modification: a nutritional sensor that modulates proteasome function. AB - The addition of O-linked beta-N-acetylglucosamine (O-GlcNAc) to serine and threonine residues is a post-translational modification of nucleocytoplasmic proteins that is thought to act in a manner analogous to protein phosphorylation. Recent work shows that many proteins of the metazoan proteasome are modified by O GlcNAc and that the level of glycosylation is responsive to the nutritional state of the cell. Moreover, increased glycosylation of the 19S (or PA700) regulatory subcomplex has been correlated with decreased proteasomal activity, suggesting a new model of proteasomal regulation. PMID- 15130577 TI - Guardian at the gate: preventing unspliced pre-mRNA export. AB - The production of a mature mRNA requires the assembly and cooperation of numerous complexes before nuclear export. The deleterious effects of intron-containing pre mRNA leakage into the cytoplasm necessitate mechanisms to prevent premature export of partially processed or unprocessed messages. A new study demonstrates that the Saccharomyces cerevisiae protein Mlp1 specifically retains intron containing pre-mRNAs in the nucleus. PMID- 15130578 TI - Why do cells need an assembly machine for RNA-protein complexes? AB - Small nuclear ribonucleoproteins (snRNPs) are crucial for pre-mRNA processing to mRNAs. Each snRNP contains a small nuclear RNA (snRNA) and an extremely stable core of seven Sm proteins. The snRNP biogenesis pathway is complex, involving nuclear export of snRNA, Sm-core assembly in the cytoplasm and re-import of the mature snRNP. Although in vitro Sm cores assemble readily on uridine-rich RNAs, the assembly in cells is carried out by the survival of motor neurons (SMN) complex. The SMN complex stringently scrutinizes RNAs for specific features that define them as snRNAs and identifies the RNA-binding Sm proteins. We discuss how this surveillance capacity of the SMN complex might ensure assembly of Sm cores only on the correct RNAs and prevent illicit, potentially deleterious assembly of Sm cores on random RNAs. PMID- 15130579 TI - Motor neurons rely on motor proteins. AB - The importance of active axonal transport to the neuron has been highlighted by the recent discoveries that mutations in microtubule motor proteins result in neurodegenerative diseases. Mutations affecting microtubule motor function have been shown to cause hereditary forms of Charcot-Marie-Tooth disease (type 2A), hereditary spastic paraplegia and motor neuron disease. Although motor neurons appear to be uniquely susceptible to defects in axonal transport, recent work has identified links between perturbations in axonal transport and the pathogenesis of other neurodegenerative diseases such as Huntington's disease and Alzheimer's disease. More broadly, cytoskeletal abnormalities might also be at the root of related disorders such as spinal muscular atrophy, supporting a key role for axonal transport in the pathogenesis of many neurodegenerative diseases. PMID- 15130580 TI - Focal adhesion and actin dynamics: a place where kinases and proteases meet to promote invasion. AB - Integrin-linked focal adhesion complexes provide the main sites of cell adhesion to extracellular matrix and associate with the actin cytoskeleton to control cell movement. Dynamic regulation of focal adhesions and reorganization of the associated actin cytoskeleton are crucial determinants of cell migration. There are important roles for tyrosine kinases, extracellular signal-regulated protein kinase/mitogen-activated protein kinase signalling, and intracellular and extracellular proteases during actin and adhesion modulation. Dysregulation of these is associated with tumour cell invasion. In this article, we discuss established roles for these signalling pathways, as well as the functional interplay between them in controlling the migratory phenotype. PMID- 15130581 TI - LRH-1: an orphan nuclear receptor involved in development, metabolism and steroidogenesis. AB - The liver receptor homolog-1 (LRH-1; NR5A2) and steroidogenic factor-1 (SF-1; NR5A1) are two orphan members of the Ftz-F1 subfamily of nuclear receptors. LRH-1 is expressed in tissues derived from endoderm, including intestine, liver and exocrine pancreas, as well as in the ovary. In these tissues, LRH-1 plays a predominant role in development, reverse cholesterol transport, bile-acid homeostasis and steroidogenesis. SF-1 expression is confined to steroidogenic tissues and the hypothalamo-pituitary-adrenal axis, where it is involved in the control of development, differentiation, steroidogenesis and sexual determination. In this article, we will review data concerning the structure, regulation and function of LRH-1. These data highlight structural similarities between LRH-1 and other Ftz-F1 members but also underscore important functional differences, assigning to LRH-1 a unique position among nuclear receptors. PMID- 15130582 TI - BAF: roles in chromatin, nuclear structure and retrovirus integration. AB - Barrier-to-autointegration factor (BAF) is an essential protein that is highly conserved in metazoan evolution. BAF binds directly to double-stranded DNA, nuclear LEM-domain proteins, lamin A and transcription activators. BAF is also a host cell component of retroviral pre-integration complexes. BAF binds matrix, a retroviral protein, and facilitates efficient retroviral DNA integration in vitro through unknown mechanisms. New findings suggest that BAF has structural roles in nuclear assembly and chromatin organization, represses gene expression and might interlink chromatin structure, nuclear architecture and gene regulation in metazoans. PMID- 15130583 TI - In a mirror dimly: tracing the movements of molecules in living cells. AB - The random movement of molecules (diffusion) is fundamental to most cellular processes, including enzymatic reactions, signalling, protein-protein interaction, as well as domain and pattern formation. Despite playing a central role, diffusion is, to a large extent, under-appreciated in the cell biology community. One reason for this is that diffusion is rather challenging to study in living cells. This article is intended to explain, at least in part, how we can go about studying diffusion of molecules in living cells, why it is important and how it provides us with important clues about biological systems. As the title 'In a mirror dimly' suggests, we do this by monitoring faint light emitted by fluorescent probes or proteins using advanced optics (e.g. mirrors) and electronics. The data are then fitted and interpreted with mathematical and physical models, providing a glimpse into the world of molecules. PMID- 15130584 TI - Blindness and brain plasticity: contribution of mental imagery? An fMRI study. AB - The purpose of this study was to study brain plasticity in the visual cortex, in six subjects totally blind from birth. The protocol we used was the same as that employed in a prior study on blindfolded sighted subjects (Brain Res., 924 (2002) 176). The production of mental images from animal names versus passive listening to abstract words, involved, in the early blind subjects as well as in the blindfolded sighted subjects of our control group, the superior occipital, inferior and superior parietal areas, premotor area, visual association. Activation foci in the somatosensory areas in the left hemisphere, as well as in the temporal and fusiform gyri were only visible in the blind subjects. The experiment, which was repeated after a short period of rest, demonstrated, this time again, predominant involvement of the dorsal pathway and activation of the primary visual area (in a region of interest). With respect to the ongoing debate on brain reorganization, our study shows that the primary visual area is activated in early blind subjects, and that activation persists in a mental imagery task involving no sensory input other than verbal instructions. PMID- 15130585 TI - Visuomotor performance in a patient with visual agnosia due to an early lesion. AB - We tested a patient with visual agnosia who had suffered severe bilateral brain damage early in life, on a series of visuomotor tasks. The broad pattern of results confirms that S.B., like the extensively tested patient D.F., shows an impressive array of preserved skills, despite his severe perceptual problems. Also like D.F., S.B. shows certain subtle visuomotor difficulties that can be related to the idea that his partially intact occipito-parietal areas are unable to benefit from interactions with the apparently severely damaged occipito temporal regions. Unlike D.F., however, he is able to make accurate discriminations of simple visual features, such as object width and orientation, albeit with very slow response times. We hypothesize that several factors such as the early onset of S.B.'s lesion and the long period since his brain lesion have allowed his brain to compensate to a degree what has been impossible in D.F., whose brain damage occurred in adulthood. This may include an element of 'rewiring' and self-monitoring of visuomotor processes that allow S.B. to achieve perceptual access to visual information processed in the dorsal stream: information that is normally only available for on-line visuomotor control. PMID- 15130586 TI - Frequency discrimination at different frequency levels as indexed by electrophysiological and behavioral measures. AB - The present study systematically compared the neural and behavioral accuracy of discriminating a frequency change ("deviant") in a repetitive tone ("standard") across a frequency range of 250-4000 Hz. The sound structure (pure sinusoidal vs. harmonically rich tones) and the magnitude of frequency change (2.5%, 5%, 10%, 20%) were also varied. The accuracy of neural frequency-change detector was determined by comparing the auditory event-related potentials (ERP) elicited by deviant and standard stimuli in the absence of attention. In a separate behavioral task, subjects were to indicate when they noticed a frequency change. The ranges of the across-subject means of ERP parameters across the conditions were: the mismatch negativity (MMN) amplitude -0.9 to -4.9 microV, latency 125 218 ms, the P3a amplitude 0.3-3.2 microV, latency 239-304 ms. The ERP latency was shortest for the standard-stimulus frequency from 1000 to 2000 Hz suggesting that automatic frequency discrimination was the most accurate in that range. The ERP latencies and amplitudes correlated with the hit rate (HR) and reaction time (RT), with highest correlation found between the MMN amplitude and the HR (r=0.8). The harmonical tones elicited MMN and P3a with shorter latencies and larger amplitudes, than did pure sinusoidal tones in all frequency bands. The results may have implication to pitch-perception theories. PMID- 15130587 TI - Category-specific modulation of inferior temporal activity during working memory encoding and maintenance. AB - Findings from neurophysiology have supported the view that visual working memory (WM) relies on modulation of activity in object-selective populations of neurons in inferior temporal cortex. Here, using event-related functional magnetic resonance imaging, we investigated whether similar mechanisms support human visual working memory encoding and maintenance processes. We identified regions in inferior temporal cortex that exhibited category-specific responses during perception of faces (fusiform face area [FFA]) or scenes (parahippocampal place area [PPA]) and investigated whether activity in these regions would be modulated by demands to actively encode and maintain faces and scenes. Results showed that independent of perceptual stimulation, the FFA and PPA exhibited greater encoding and maintenance-related activity when their favored stimulus was relevant to the recognition task. In contrast, maintenance-related activity in the dorsolateral prefrontal cortex (PFC) was modulated by memory load, regardless of the type of information that was task relevant. These results are consistent with the view that visual working memory encoding and maintenance processes are implemented through modulation of inferior temporal activity by prefrontal cortex. PMID- 15130588 TI - The cerebellum and decision making under uncertainty. AB - This study aimed to identify the neural basis of probabilistic reasoning, a type of inductive inference that aids decision making under conditions of uncertainty. Eight normal subjects performed two separate two-alternative-choice tasks (the balls in a bottle and personality survey tasks) while undergoing functional magnetic resonance imaging (fMRI). The experimental conditions within each task were chosen so that they differed only in their requirement to make a decision under conditions of uncertainty (probabilistic reasoning and frequency determination required) or under conditions of certainty (frequency determination required). The same visual stimuli and motor responses were used in the experimental conditions. We provide evidence that the neo-cerebellum, in conjunction with the premotor cortex, inferior parietal lobule and medial occipital cortex, mediates the probabilistic inferences that guide decision making under uncertainty. We hypothesise that the neo-cerebellum constructs internal working models of uncertain events in the external world, and that such probabilistic models subserve the predictive capacity central to induction. PMID- 15130589 TI - Modulation of right motor cortex excitability without awareness following presentation of masked self-images. AB - The neural substrates of self-awareness have been studied with a variety of neurophysiological and behavioral tools. In the present study, unconscious modulation of corticospinal excitability following presentation of self-images was probed with transcranial magnetic stimulation (TMS). TMS-induced motor evoked potentials (MEP) were collected from the contralateral first dorsal interosseus (FDI) muscle while subjects viewed masked pictures of their own face. MEP amplitudes were compared to those obtained when pictures of strangers were masked. Masked self-images induced a relative increase in corticospinal excitability when TMS was applied to the right primary motor cortex. These results demonstrate the utility of TMS to probe unconscious processing and support the notion of hemispheric asymmetry in the processing of self-images. PMID- 15130590 TI - Selective spatial attention to left or right hand flutter sensation modulates the steady-state somatosensory evoked potential. AB - Steady-state somatosensory evoked potentials (SSSEPs) were recorded from the scalp of human subjects elicited by 20 and 26 Hz mechanical vibrations applied simultaneously to the index finger of the left (20 Hz) and right hand (26 Hz). Subjects were instructed to attend to the flutter vibration at one finger while ignoring the other finger and to detect rare target events at the to-be-attended finger. The amplitude of the frequency coded SSSEP elicited by the attended vibration was significantly enlarged when attention was focused at the respective finger. This amplitude enhancement with attention was most prominent over fronto central electrode locations contralateral to the attended finger. This is the first report to show the attentional modulation of the SSSEP amplitude in humans, suggesting an enhancement of neural responses in the sense of flutter with attention. The findings will open a new approach for studying the neural mechanisms of sustained selective attention in somatosensation. PMID- 15130591 TI - Emotion-attention network interactions during a visual oddball task. AB - Emotional and attentional functions are known to be distributed along ventral and dorsal networks in the brain, respectively. However, the interactions between these systems remain to be specified. The present study used event-related functional magnetic resonance imaging (fMRI) to investigate how attentional focus can modulate the neural activity elicited by scenes that vary in emotional content. In a visual oddball task, aversive and neutral scenes were presented intermittently among circles and squares. The squares were frequent standard events, whereas the other novel stimulus categories occurred rarely. One experimental group [N=10] was instructed to count the circles, whereas another group [N=12] counted the emotional scenes. A main effect of emotion was found in the amygdala (AMG) and ventral frontotemporal cortices. In these regions, activation was significantly greater for emotional than neutral stimuli but was invariant to attentional focus. A main effect of attentional focus was found in dorsal frontoparietal cortices, whose activity signaled task-relevant target events irrespective of emotional content. The only brain region that was sensitive to both emotion and attentional focus was the anterior cingulate gyrus (ACG). When circles were task-relevant, the ACG responded equally to circle targets and distracting emotional scenes. The ACG response to emotional scenes increased when they were task-relevant, and the response to circles concomitantly decreased. These findings support and extend prominent network theories of emotion-attention interactions that highlight the integrative role played by the anterior cingulate. PMID- 15130592 TI - Enhanced neural activity in response to dynamic facial expressions of emotion: an fMRI study. AB - Dynamic facial expressions of emotion constitute natural and powerful media of communication between individuals. However, little is known about the neural substrate underlying the processing of dynamic facial expressions of emotion. We depicted the brain areas by using fMRI with 22 right-handed healthy subjects. The facial expressions are dynamically morphed from neutral to fearful or happy expressions. Two types of control stimuli were presented: (i) static facial expressions, which provided sustained fearful or happy expressions, and (ii) dynamic mosaic images, which provided dynamic information with no facial features. Subjects passively viewed these stimuli. The left amygdala was highly activated in response to dynamic facial expressions relative to both control stimuli in the case of fearful expressions, but not in the case of happy expressions. The broad region of the occipital and temporal cortices, especially in the right hemisphere, which included the activation foci of the inferior occipital gyri, middle temporal gyri, and fusiform gyri, showed higher activation during viewing of the dynamic facial expressions than it did during the viewing of either control stimulus, common to both expressions. In the same manner, the right ventral premotor cortex was also activated. These results identify the neural substrate for enhanced emotional, perceptual/cognitive, and motor processing of dynamic facial expressions of emotion. PMID- 15130593 TI - Decay of prism aftereffects under passive and active conditions. AB - In prism adaptation, subjects adapt to new visuospatial coordinates imposed by wedge prisms that laterally displace the visual field. During this process, subjects develop and store new visuomotor coordinates in order to compensate for the displacement of visual stimuli. After the prisms are removed, subjects show an aftereffect in the opposite direction of the original perturbation. The aftereffect is a manifestation of the recently stored information. In the present article, we were interested in studying the properties of the aftereffect. Specifically, we investigated the fate of the aftereffect under active conditions with motor reafferences but without visual input, and during passive conditions without visual or motor reafferences. The results in the motor active condition show that motor reafference (proprioceptive or corollary discharge information) led to a faster, but incomplete, aftereffect decay. The results in the passive condition show a bimodal aftereffect behavior, with a fast decay within the initial minutes, followed by a sustained aftereffect up to 20 min later. These data suggests that two different memory processes may contribute to the aftereffect, one showing a fast decay mainly within 1 min, and another that shows a stable endurance for more than 20 min. PMID- 15130594 TI - Event-related potentials elicited during parsing of ambiguous relative clauses in Spanish. AB - Previous behavioural studies in Spanish have found a significant preference for attaching relative clauses preceded by a complex NP (N1 of N2 RC) to the first noun phrase. In the present study, we used event-related potentials (ERPs) to help identify the nature of these processes by directly comparing ERPs to temporary ambiguous sentences containing relative clauses that were finally consistent with either high or low attachment resolution. The larger amplitude of the P600 effect for the low attachment condition suggests that high attachment was the preferred strategy. The P600 effect was widely distributed in the 500-700 ms window, including frontal areas, while the distribution was mainly posterior in the 700-1000 ms window. The results indicate that high attachment is the parsing strategy Spanish readers use for this type of ambiguity and suggest that the P600 may not be a monolithic effect. PMID- 15130595 TI - The similarity of brain morphology in healthy monozygotic twins. AB - Aim of the study was to investigate the degree of similarity of twin brains with respect to their outer shape and gyrification. High resolution MRI was obtained from 26 healthy monozygotic twins (MZ) and three-dimensional renderings of the brains were generated. Similarity was rated by human investigators and by computer analysis. Three different image types were analyzed: whole-brain views, silhouettes and a bird's-eye view of a segment showing the central region. For each of the three image types, 13 tasks (identifying the related twin-pair out of a set of five brains) had to be solved by the human raters. For whole brain, views and silhouettes 66/91(p<0.005) and for segment views 44/91 (p<0.02) correct identifications were made. Using cross correlation coefficients, the computer based analysis as well significantly often identified related twins. Again correct identification was more likely based on whole-brain views and silhouettes than on segment views of the central region. In conclusion, we found that overall brain shape is probably strongly influenced by genetic effects but the variation in sulcal and gyral patterns is also affected by non-genetic influences to a considerable extent. PMID- 15130596 TI - Effect of chlorpyrifos on efflux transporter gene expression and function in Caco 2 cells. AB - The effect of chlorpyrifos (CPF) and its metabolite, chlorpyrifos-oxon (CPO), on multidrug resistance-1 (MDR1) gene expression and efflux transporter function in Caco-2 cells was determined. The effect of CPF and CPO on gene expression in Caco 2 cells was tested as a function of time using RT-PCR and competitive PCR (compPCR) techniques. The RT-PCR results depicted a maximal effect of CPF exposure on MDR1 expression at 8 h, which decreased at 24 h. Studies with CPO displayed an initial increase in expression at 4 h only. The compPCR assays were conducted with the CPF-treated group to quantify the changes in gene expression levels. The compPCR data confirmed and quantitated the results from the time course study using semiquantitative RT-PCR. In addition to the gene expression studies, changes in efflux transporter function were investigated using Caco-2 cells grown on semipermeable membranes in Transwell plates. The permeability of verapamil was determined in cells treated for 8 h with CPF. Efflux ratios demonstrated that verapamil was effluxed at a higher rate from the CPF-treated cells as compared to the control group, confirming the inductive action of CPF on transporter function. These results suggest that CPF has the potential to modulate the bioavailability of drugs via changes in expression and function of membrane efflux transporters. PMID- 15130597 TI - Impact of antiseptics on radical metabolism, antioxidant status and genotoxic stress in blood cells: povidone-iodine versus octenidine dihydrochloride. AB - No sufficient data are available of the of antiseptics' influence on human blood cells. Effects of two antiseptics, povidone-iodine (PVD-I) versus octenidine dihydrochloride (OD), were tested on antioxidant status, radical formation, antioxidant defence enzymes and genotoxic stress in blood cells, in vitro. Human blood was taken by venipuncture, enriched with PVD-I or OD (0.0001-20% final concentration) and incubated at 37 degrees C between 30 and 120 min. alpha Tocopherol was assessed in erythrocytes and granulocytes. Superoxide-dismutase (SOD) and glutathione (GSH) were determined in erythrocytes, the total anti oxidative capacity (TAC) and malondialdehyde (MDA) in their ghosts. In granulocytes status of hydrogen peroxide (H(2)O(2)), superoxide anions and MDA was observed. Genotoxic stress was determined by counting sister chromatide exchanges (SCE) in lymphocytes after enrichment within 0.05-0.4% of antiseptics. Based on all biomarker tested, concentrations up to 0.05% incubated for 30 min did not affect cell metabolism. 1% and 10% PVD-I reduced the activity of SOD ( 40%), GSH (-62%) and the content of alpha-tocopherol more than OD (p<0.05). No significant differences between the antiseptics were observed for TAC and MDA. H(2)O(2) and superoxide anions were significantly reduced after the 10% addition for both substances independent on the exposure. Without having changes in lipid oxidation, the reduction of antioxidative defence mechanisms must be due to the oxidation caused by the antiseptics, mainly PVD-I. An increased SCE rate was neither observed with PVD-I nor with OD within an enrichment with 0.05-0.4%. Higher concentrations (1% and more) could not be tested on SCE formation because they caused cell bursts. The results presented indicate that concentrations up to 0.05% incubated for 30 min are safe for exposing blood cells of healthy subjects. PMID- 15130598 TI - Cytotoxic effect and role of exogenous antioxidants in carpet dust mediated toxicity in rat hepatocytes in vitro. AB - Carpet industries bear a great deal of economic and commercial significance in India. In order to safe guard the workers against the health hazards caused by dust in their occupational environment; it necessitates studying the biological importance of these dusts. The present study was designed to investigate the toxicity of carpet dust (knotted and tuffted) on isolated rat hepatocytes. The hepatocytes were isolated by collagenase perfusion method and cells were incubated with different concentration of carpet dust (100-5000 microg/10(6) cells) with various time (30-180 min) intervals. An exogenous antioxidant vitamin E also used to find out the role of antioxidants and free radical production in carpet dust mediated toxicity. Cell viability by trypan blue exclusion and leakage of enzyme lactate dehydrogenase (LDH) were determined. Reduced glutathione (GSH), formation of thiobarbituric acid reactive substance (TBARS) were also measured. A significant decrease in the cell viability was observed after 60, 180 min upon incubation with tuffted carpet dust, while knotted carpet dust caused a significant decrease in the viability after 180 min. LDH leakage was parallel to the cell viability. Thiobarbituric acid reactive substance was significantly increased at 30 and 60 min with carpet dust treated hepatocytes. Dust at 1000 and 5000 microg dose level showed significantly increased formation of TBARS at 30 min incubation. However, when hepatocytes were co-incubated with carpet dust and Vit-E (10, 15 microM), a significant decrease in LDH release and TBARS production was observed while 15 microM Vit-E showed an enhanced protection than 10 microM Vit-E treated hepatocytes. The effect of carpet dust on cell viability, LDH leakage, TBARS production, GSH depletion was time and dose dependent. Moreover, we observed that tuffted carpet dust causes greater effect than knotted one on the above mentioned parameters. Our studies also revealed that Vit-E in culture media diminishes the carpet dust mediated toxicity. PMID- 15130599 TI - Effects of the pesticides prochloraz and methiocarb on human estrogen receptor alpha and beta mRNA levels analyzed by on-line RT-PCR. AB - Exposure to endocrine disrupters such as dioxins, PCBs and certain pesticides are suspected to affect human reproductive health. We have analyzed the effect of the currently used pesticides prochloraz and methiocarb on the estrogen receptor (ER)alpha and beta mRNA levels in parallel with the natural ligand, 17beta estradiol (E2). Using the highly sensitive on-line RT-PCR technique we were able to quantify the ERalpha and ERbeta mRNA levels in the human breast cancer cell line, MCF7-BUS. Upon exposure with E2 or prochloraz a down regulation of ERalpha and ERbeta mRNAs was observed after 48 h of treatment. Co-treatment with the ER antagonist ICI 182,780 abolished these mRNA down regulations. Western blot analyses elicited a decreased ER protein level after 3 h of exposure with prochloraz but after 24 h the ERalpha protein level had recovered to basal level. Methiocarb exposure had no effect on the ERalpha mRNA level, whereas an increase in the ERbeta mRNA level was observed after 3 h of exposure. Our study demonstrates that like E2, prochloraz had the potential to down regulate the expression of ERalpha and ERbeta mRNAs as well as the ERalpha protein level in MCF7-BUS cells. PMID- 15130600 TI - The biflavonoid, amentoflavone degrades DNA in the presence of copper ions. AB - Previous reports from this laboratory have shown that flavonoids including apigenin are capable of inducing oxidative DNA cleavage in the presence of copper ions. In the present report, we have examined the ability of amentoflavone, a biflavonoid which is a dimer of apigenin, to catalyze the degradation of DNA. Amentoflavone was found to degrade calf thymus DNA in the presence of Cu(II) at a rate almost twice that of apigenin. Amentoflavone was also shown to reduce Cu(II) to Cu(I) and to generate hydroxyl radicals in the presence of copper ions. In the presence of Cu(II), the absorption spectrum of amentoflavone undergoes a shift and a quenching effect indicating that the biflavonoid is capable of binding to copper ions. Amentoflavone and apigenin were isolated from Cycas rumphii and Trifolium alexandrinum, respectively. The results are discussed in relation to the putative chemopreventive mechanism of amentoflavone. PMID- 15130601 TI - Chloramphenicol succinate, a competitive substrate and inhibitor of succinate dehydrogenase: possible reason for its toxicity. AB - From our previous study [Eur. J. Clin. Pharmacol. 56 (2000) 405] we hypothesized that chloramphenicol succinate (CAPS) may be a competitive substrate for succinate dehydrogenase (SDH). It may be oxidized by SDH to release chloramphenicol (CAP), which may inhibit SDH by feed back mechanism. The present ex-vivo/in vitro study was aimed to investigate this possibility by using human tissues (bone marrow and liver samples) and animal tissues (rat liver and kidney). The effect of different SDH activators and specific inhibitors was studied on CAPS metabolism by SDH. The metabolites and reduction products were detected by using HPLC. In marrow samples, CAPS was slowly oxidized to form CAP. The formation of CAP (oxidation product) was enhanced by FAD and low malonate and inhibited by high malonate and 3-NPA. Similar results were obtained with mitochondria from human and rat tissues. These studies suggest that CAPS could be a competitive oxidative substrate and the metabolite CAP could be an inhibitor at the reduction site. Therefore, SDH could be a target molecule responsible for CAPS induced toxicity. PMID- 15130602 TI - Morphine-promoted survival of CEMx174 cells in early stages of SIV infection in vitro: involvement of the multiple molecular mechanisms. AB - Progression of HIV infections to AIDS is a complex process and it differs considerably among individuals infected with HIV, influenced by both genetic and environmental factors. Opiates have been implicated to be a cofactor in HIV infections leading to AIDS. However, little is known about the molecular mechanisms involved in the effects of opioids on HIV infected immune cells. Cell cycle analysis was carried out by flow cytometry, the phosphorylation of mitogen activated protein kinases ERK1 and ERK2 was detected by Western blotting assay, and changes of calcium concentration were monitored by scanning intracellular fluorescence intensity. In response to the treatment with morphine, SIV-infected cells were accumulated in G1 phase. Morphine increased the content of intracellular calcium in a time-dependent manner. In addition, morphine also elevated the levels of PKC activity and phosphorylated ERK1/2. Therefore, it is implicated that the calcium-PKC-MAPK cascade is involved in morphine-prolonged survival of SIV-infected cells in the early stages of virus infection. PMID- 15130603 TI - Evaluation of toxic/protective effects of the essential oil of Salvia officinalis on freshly isolated rat hepatocytes. AB - For this study the essential oil (EO) of sage (Salvia officinalis L.) was isolated from air-dried vegetative aerial parts of the plants by hydrodistillation and analysed by GC and GC-MS. A total yield of 12.07 mg of EO per g of plant dry mass was obtained and more than 50 compounds identified. The major compounds were cis-thujone (17.4%), alpha-humulene (13.3%), 1,8-cineole (12.7%), E-caryophyllene (8.5%) and borneol (8.3%). The EO fraction of sage tea was also isolated by partition with pentane and the respective components identified. The toxic and antioxidant protective effects of S. officinalis EO were evaluated on freshly isolated rat hepatocytes. Cell viability (LDH leakage), lipid peroxidation and glutathione status were measured in experiments undertaken with cells (suspensions of 1 x 10(6) cells per millilitre) exposed to EO alone (toxicity of the EO;t-BHP as positive control); and with cells exposed to EO and an oxidative compound (t-BHP) together (in EO protection evaluation; quercetin as positive control) for 30 min. The results show that the EO is not toxic when present at concentrations below 200 nl/ml; it was only at 2000 nl EO/ml that a significant LDH leakage and GSH decrease were observed indicating cell damage. In the range of concentrations tested, the EO did not show protective effects against t-BHP-induced toxicity. PMID- 15130604 TI - Cytotoxicity, inhibition of DNA and protein syntheses and oxidative damage in cultured cells exposed to zearalenone. AB - Mycotoxins are toxic metabolites of various fungi commonly found in feed and foodstuff and can cause very serious health problems in animals as well as in humans. Zearalenone (ZEN), a mycotoxin produced by various Fusarium species has several adverse effects. Indeed, ZEN has strong estrogenic activity associated with hyperestrogenism and several physiological alterations of the reproductive tract. Moreover, ZEN was shown to be hepatotoxic, haematotoxic, immunotoxic and genotoxic. The exact mechanism of ZEN toxicity is not completely established. The observed strong estrogenic effect of ZEN resulting from its competition with 17beta-estradiol in the binding to estrogen receptors is generally considered to underline most toxic effects of ZEN, but estrogenic activity alone cannot explain the diverse and apparent adverse effects. The objective of the present study was to determine the involvement of other possible mechanisms in ZEN induced toxicity. Cytotoxicity, cell cycle perturbation, inhibition of protein and DNA synthesis as well as the presumed later marker of oxidative stress, malondialdehyde, were monitored in Vero and Caco-2 cells exposed to ZEN. Our results showed that ZEN reduces cell viability correlated to cell cycle perturbation, inhibits protein and DNA syntheses and increases MDA formation in both cell lines in concentration-dependant manner. We assumed that cytotoxicity and oxidative damage are additional mechanisms of ZEN mediated toxicity. PMID- 15130605 TI - Organotellurium compound toxicity in a promyelocytic cell line compared to non tellurium-containing organic analog. AB - Two substituted aryl organotellurium compounds and a tellurium-free analog of one of these were evaluated for in vitro cytotoxicity using human promyelocytic (HL 60) cells as an experimental system. Both tellurium-containing toxicants (2,2(') dimethoxydiphenyl ditelluride and 2,2(')-diamino-3,3('),5,5(') tetramethyldiphenyl ditelluride) were cytotoxic at concentrations as low as 5 x 10(-6) M and the dimethoxydiphenyl compound produced significant numbers of apoptotic cells at a concentration of only 1 x 10(-6) M after 8 h. Data indicate that 2,2(')-dimethoxydiphenyl ditelluride and 2,2(')-diamino-3,3('),5,5(') tetramethyldiphenyl ditelluride induce apoptosis in both a time and dose dependent manner; however, 2,2(')-dimethoxybiphenyl, structurally comparable to the first of these but without the tellurium bridge, did not produce apoptosis under the concentrations and time course studied. Therefore the telluride moiety was apparently an important factor in the apoptotic effect. PMID- 15130606 TI - Flow cytometric methods used as screening tests for basal toxicity of chemicals. AB - Aim of the present study was to evaluate the suitability of flow cytometry to test in vitro effects of toxicants. Flow cytometry offers the possibility to study several parameters simultaneously, e.g. cell cycle modulation, apoptosis and necrosis within the same cell culture. The effects of six compounds (acetaminophen=AAP, isoniazid=INH, digoxin, malathion, paraquat and 2,4 dichlorophenoxy acetic acid=2,4-D) on cell cycle were investigated in HepG2 cells and the induction of apoptosis/necrosis was analyzed by a spectrum of flow cytometric assays in HepG2, AAH-1 and YAC-1 cells. Early indicators of apoptosis- loss of mitochondrial membrane polarization--as well as later events of the apoptotic process--annexin V binding and DNA fragmentation--were studied. The phases of the cell cycle and the occurrence of a sub-G(0) peak of apoptotic cells were determined with propidium iodide staining. The present investigation demonstrated good correlations between results obtained by flow cytometric analyses and the IC50 data of the MEIC (=Multicenter Evaluation of In Vitro Cytotoxicity) study. Regarding the short time required for the tests, the possibility of investigating several parameters of cytotoxicity simultaneously and the ease of performance, flow cytometric analyses are well suited for the pre screening for toxic effects of chemicals. PMID- 15130607 TI - Moderate skin sensitizers can induce phenotypic changes on in vitro generated dendritic cells. AB - In the present study, we analyzed the phenotypic alterations induced by several allergens on immature dendritic cells (DC), with the aim to develop a potential in vitro alternative for predicting the sensitizing potential of chemicals. DC were generated from human monocytes cultured in the presence of GM-CSF, IL-4 and TGF-beta1 and treated for 2 or 4 days with different chemicals. Surface marker expression (HLA-DR, CD1a, CD40, CD54, CD83, CD86, CCR7 and E-cadherin) was analyzed by flow cytometry. Results showed that a 2-day treatment with the representative allergens DNCB and NiSO(4) induced significant changes of most antigens while other chemicals such as balm of Peru (strong allergen), kathon (moderate allergen), cinnamic aldehyde (mild allergen) or the irritant SLS had no significant effect. In contrast, the 4-day treatment with allergens substantially improved the results. Indeed, despite a large variability according to the donors, the number of modified antigens was significantly higher with all the tested chemicals, except kathon, as compared to that observed with the irritant SLS. The present study indicates that, in this model, the screening of mild or moderate allergens requires both the consideration of many antigens and a prolonged time of incubation with the chemicals. PMID- 15130608 TI - In vitro assessment of copper-induced toxicity in the human hepatoma line, Hep G2. AB - Copper, though essential, is highly toxic when present in excess, as in Wilson disease, a genetic disorder of hepatic copper metabolism. We hypothesized that mitochondria are a major target of copper-induced cytotoxicity in Wilson disease. We used the human hepatoma line Hep G2 to examine copper-mediated cytotoxicity and three different methods to assess organelle damage: MTT assay (mitochondria), neutral red (NR; lysosomes) and Trypan blue exclusion assay (TB; plasma membrane). For all assays, cells at approximately 60% confluence in microtitre plates were incubated with CuCl(2) (concentration range: 50-100-150-200 microM) for 24 or 48 h. Results were expressed as percent of untreated control. At 24 h, cytotoxicity as detected by NR assay was significantly higher at all concentrations of copper than for MTT or TB ( p<0.005 at all concentrations). Cytotoxicity as detected by MTT was higher than that detected by TB at all concentrations except at 200 microM (p<0.05 for 50 microM, p<0.005 for 100 microM, p = 0.001 for 150 microM). Results at 48 h were similar (NR versus others: p <0.001 MTT versus TB: NS except at 150 microM where p<0.01). We investigated reactive oxygen species (ROS) production in copper-associated hepatocytoxicity by incubating sub-confluent cells with 2('),7(') dichlorodihydrofluorescein diacetate dye plus copper (concentration range: 0-200 microM) for 1-1.5 h. Copper, but not zinc, produced significant increases in ROS (p<0.001). In summary, Hep G2 lysosomes appeared more susceptible to Cu-mediated damage than mitochondria; the cell membrane was highly resistant to damage. PMID- 15130609 TI - Discriminative power of an assay for automated in vitro screening of teratogens. AB - Screening for potential teratogenicity of 20 test compounds was performed using a computerised microscope workstation for determination of cytotoxicity, proliferation and morphology of fibroblastoid murine L929-cells. The test compounds, which were divided into four classes according to teratogenicity were: 5-bromo-2(')-deoxyuridine, 6-aminonicotinamide, acrylamide, boric acid, D-(+) camphor, dimethadione, dimethyl phthalate, diphenhydramine, hydroxyurea, isobutyl ethyl-valproic acid, lithium chloride, methyl mercury chloride, methotrexate, methoxyacetic acid, penicillin G, all-trans-retinoic acid, pentyl-4-yn-valproic acid, saccharin, salicylic acid and valproic acid. All compounds, with the exception of dimethadione inhibited proliferation in a linear dose-dependent manner, and there were statistically significant compound class-dependent differences between the IC(50)-values for the compounds (p<0.0374), the strongest teratogens being the most potent. Furthermore, the average efficacies (maximum relative change) for 10 parameters describing cell morphology exhibited statistically significant compound class-dependent differences (p<0.0001), the class I and II compounds exhibiting significantly lower efficacies than the class III and IV compounds (p<0.01). Thus, test compounds affected both the proliferation and morphology of L-cells in manner demonstrating a general relationship with the teratogenic potency of the compounds. However, the moderate teratogens dimethadione and lithium chloride only had minor effects on the morphology and proliferation of the cells whereas the non-teratogen diphenhydramine had effects on both proliferation and morphology comparable to the strong teratogens. PMID- 15130610 TI - Phosphorylation of hepatocyte growth factor receptor and epidermal growth factor receptor of human hepatocytes can be maintained in a (3D) collagen sandwich culture system. AB - In vitro culture models that employ human liver cells could be potent tools for predictive studies on drug toxicity and metabolism in the pharmaceutical industry. However, an adequate receptor responsiveness is necessary to allow intracellular signalling and metabolic activity. We tested the ability of three dimensionally arranged human hepatocytes to respond to the growth factors hepatocyte growth factor (HGF) or epidermal growth factor (EGF). Isolated adult human hepatocytes were cultivated within a three-dimensional collagen gel (sandwich) or on a two-dimensional collagen matrix. Cells were treated with HGF or EGF and expression and phosphorylative activity of HGF receptors (HGFr, c-met) or EGF receptors (EGFr) were measured by flow cytometry and Western blot. Increasing HGFr and EGFr levels were detected in hepatocytes growing two dimensionally. However, both receptors were not activated in presence of growth factors. In contrast, when hepatocytes were plated within a three-dimensional matrix, HGFr and EGFr levels remained constantly low. However, both receptors became strongly phosphorylated by soluble HGF or EGF. We conclude that cultivation of human hepatocytes in a three-dimensionally arranged in vitro system allows the maintenance of specific functional activities. The necessity of cell dimensionality for HGFr and EGFr function should be considered when an adequate in vitro system has to be introduced for drug testing. PMID- 15130611 TI - Identification of in vitro protein biomarkers of idiosyncratic liver toxicity. AB - Drug-induced idiosyncratic hepatotoxicity continues to be an important safety issue for the pharmaceutical industry. This toxicity is due, in part, to the limited predictive nature of current pre-clinical study systems. A hypothesis was formed that treatment of existing in vitro hepatocyte cultures with drugs clinically linked to idiosyncratic hepatotoxicity would result in the release of extracellular protein biomarkers indicative of liver toxicity. To test this hypothesis, a combination of proteomic and immunological techniques were used to first identify, and subsequently verify, components of the protein-laden conditioned culture media from immortalized human hepatocytes which overexpressed cytochrome p450 3A4. These cells were treated separately with seven individual compounds made up of a combination of thiazolidinedione and l-tyrosine PPARgamma agonists and HIV protease inhibitors, plus a vehicle control (dimethyl sulfoxide). For each drug class, clinically determined hepatotoxic and non hepatotoxic compounds were compared. Two proteins, BMS-PTX-265 and BMS-PTX-837, were reproducibly and significantly increased in the conditioned media from cells treated with each of the toxic compounds as compared to media from cells treated with the non-toxic compounds (and vehicle). This result supported the hypothesis, and so a series of successive assays (western blots and enzyme linked immunosorbent assays) were used to measure the response of these two proteins as a function of an expanded set of 20 compounds. For all 20 drugs, elevations of BMS-PTX-265 correlated exactly with the known safety profile; whereas changes in BMS-PTX-837 correctly predicted the safety profile in 19 of 20 drugs (one false negative). In summary, the data supports both the pre-clinical in vitro method as a means to identify new biomarkers of liver toxicity, as well as the validity of the biomarkers themselves. PMID- 15130612 TI - Combined effects of Matrigel and growth factors on maintaining undifferentiated murine embryonic stem cells for embryotoxicity testing. AB - Undifferentiated, murine embryonic stem (mES) cells have shown promise as a substrate for identifying embryotoxic chemicals and for studying mechanisms of early developmental injury. However, long-term maintenance of mES cells in an undifferentiated state is problematic. The present study evaluates the combination of Matrigel matrix and three growth factors for this purpose. Biomarkers of mES cell pluripotency, apoptosis, chromosome number and cardiomyocyte differentiation were monitored over 119 population doublings. D3 mES cells retained undifferentiated characteristics, including sustained expression of alkaline phosphatase and stage specific embryonic antigen-1 (SSEA 1) and continued transcription of Pou5f1 (Oct-4). Cell viability remained at > or=95% and population-doubling times averaged 14.3 h over 10 weeks of observation. Caspase-3 activation, a marker of cellular death by apoptosis, was measured in early- and late-passage mES cells. Early-passage cells showed dose responsive caspase-3 activation following exposure to sodium arsenite, whereas caspase-3 activation of late-passage cells dropped to background levels at toxicant dosages above 50 ppb. Aneuploidy and impaired differentiation into beating cardiomyocytes were noted for late-passage mES cells. Matrigel, combined with growth factors, may sustain undifferentiated mES cells. However, aneuploidy, reduced caspase-3 activation, and inability to differentiate suggests further modifications to the culture system may be needed for long-term propagation of cells for embryotoxicity endpoints. PMID- 15130613 TI - The thick and thin of LASIK flaps. PMID- 15130614 TI - Phakonit: Incisions and use of a pressurized inflow system. PMID- 15130616 TI - Induced astigmatism after laser in situ keratomileusis. PMID- 15130618 TI - USST and no cycloplegic drops before cataract surgery. PMID- 15130619 TI - Control and consideration of wavefront aberrometers. PMID- 15130620 TI - Fourier analysis. PMID- 15130621 TI - Consultation section. Refractive surgical problem. PMID- 15130628 TI - Management of lens-iris diaphragm retropulsion syndrome during phacoemulsification. AB - Lens-iris diaphragm retropulsion syndrome (LIDRS) occurs more often than recognized during small-incision phacoemulsification. This syndrome requires an infusion of fluid into the anterior chamber and is characterized by posterior displacement of the lens-iris diaphragm, marked deepening of the anterior chamber, posterior iris bowing, pupil dilation, and often significant patient discomfort. Using microendoscopy, we have observed that LIDRS is essentially a reverse pupillary block. We describe a surgical technique to mechanically break the iridocapsular block to restore normal chamber depth, relieve patient discomfort, and allow the surgeon to proceed safely with phacoemulsification. PMID- 15130629 TI - Suture burial technique in scleral fixation. AB - The end of a scleral fixation suture may erode through the conjunctiva and become exposed. To avoid this, a scleral flap, autologous cornea, dura mater, or fascia lata patches have been used to cover the ends, and rotation of the knot into the tissue has been reported. An alternative technique to bury the end of the suture is described. PMID- 15130630 TI - Continuous curvilinear capsulorhexis with use of an endoscope. AB - We describe a technique for creating a continuous curvilinear capsulorhexis (CCC) using an endoscope. Three 2.0 mm limbal incisions 120 degrees apart were made in 4 cadaver eyes. The anterior capsule was stained with trypan blue 0.1% (Vision Blue). Under endoscopic visualization, a CCC was attempted in each of the 4 globes. It was completed in 1 of them. Endoscopy provides a safe and effective tool to visualize the anterior chamber and perform a CCC when there is a poor view through the cornea. PMID- 15130631 TI - Flap thickness accuracy: comparison of 6 microkeratome models. AB - PURPOSE: To determine the flap thickness accuracy of 6 microkeratome models and determine factors that might affect flap thickness. SETTING: Magill Research Center for Vision Correction, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: This multicenter prospective study involved 18 surgeons. Six microkeratomes were evaluated: AMO Amadeus, Bausch & Lomb Hansatome, Moria Carriazo-Barraquer, Moria M2, Nidek MK2000, and Alcon Summit Krumeich-Barraquer. Eyes of 1061 consecutive patients who had laser in situ keratomileusis were included. Age, sex, surgical order (first or second cut), keratometry (flattest, steepest, and mean), white-to-white measurement, laser used, plate thickness, head serial number, blade lot number, and occurrence of epithelial defects were recorded. Intraoperative pachymetry was obtained just before the microkeratome was placed on the eye. Residual bed pachymetry was measured after the microkeratome cut had been created and the flap lifted. The estimated flap thickness was determined by subtraction (ie, mean preoperative pachymetry measurement minus mean residual bed pachymetry). RESULTS: A total of 1634 eyes were reviewed. Sex distribution was 54.3% women and 45.7% men, and the mean age was 39.4 years +/- 10.6 (SD). In addition, 54.5% of the procedures were in first eyes and 45.5%, in second eyes. The mean preoperative pachymetry measurement was 547 +/- 34 microm. The mean keratometry was 43.6 +/- 1.6 diopters (D) in the flattest axis and 44.6 +/-1.5 D in the steepest axis. The mean white to-white measurement was 11.7 +/- 0.4 mm. The mean flap thickness created by the devices varied between head designs, and microkeratome heads had significant differences (P<.05). Factors that explained 78.4% of the variability included microkeratome model, plate thickness, mean preoperative pachymetry, Kmin, surgery order, head serial number, blade lot number, and surgeon. Factors such as age, sex, Kmax, Kaverage, white to white, and laser had no significant correlation to flap thickness. CONCLUSIONS: The results demonstrated variability between the 6 microkeratome models. Device labeling did not necessarily represent the mean flap thickness obtained, nor was it uniform or consistent. Thinner corneas were associated with thinner flaps and thicker corneas with thicker flaps. In addition, first cuts were generally associated with thicker flaps when compared to second cuts in bilateral procedures. PMID- 15130632 TI - Posterior capsule opacification after implantation of CeeOn Edge 911A, PhacoFlex SI-40NB, and AcrySof MA60BM lenses: one-year results of an intraindividual comparison multicenter study. AB - PURPOSE: To perform an intraindividual comparison of posterior capsule opacification (PCO) with 2 foldable intraocular lenses (IOLs) and a foldable acrylic IOL 1 year after in-the-bag implantation. SETTINGS: Seven German ophthalmology centers. METHODS: In an open prospective randomized multicenter study, each center intraindividually compared a high-refractive-index, sharp edged optic silicone IOL (CeeOn Edge 911A, Pharmacia) with a high-refractive index, round-edged optic silicone IOL (PhacoFlex SI-40NB, Allergan) or a sharp edged optic acrylic IOL (AcrySof MA60BM, Alcon). Of 288 randomized patients, 247 had standard phacoemulsification with in-the-bag IOL implantation in both eyes by the same surgeon. One eye of each patient received a CeeOn Edge IOL and the fellow eye, an AcrySof or PhacoFlex IOL. A morphologic evaluation of PCO was performed using the Evaluation of Posterior Capsule Opacification (EPCO) system 1 to 2 weeks and 11 to 14 months after surgery. The digital pictures were evaluated by an independent investigator who was blind to the type of IOL. Intraindividual differences in EPCO scores were statistically evaluated by a 1-sided binomial test at an alpha-level of 5%. RESULTS: One year after surgery, 127 patients with the AcrySof IOL and 102 patients with the PhacoFlex IOL in the control eye were reexamined. Functional results, safety, and handling were not significantly different between the 3 IOLs. All reexamined eyes had a very low PCO grade. The EPCO values revealed less PCO in eyes with the CeeOn Edge IOL than in eyes with the AcrySof or PhacoFlex IOL, but the difference was not statistically significant. A neodymium:YAG laser capsulotomy was performed in 1 eye with a CeeOn Edge IOL, 1 eye with an AcrySof IOL, and 2 eyes with a PhacoFlex IOL. CONCLUSIONS: The EPCO PCO grade was low 1 year after implantation of CeeOn Edge 911A, PhacoFlex SI-40NB, and AcrySof MA60BM IOLs; there was no statistically significant difference between the IOLs. The impact of IOL material and edge design on PCO development might be relevant in a long-term follow-up of this study. PMID- 15130633 TI - Improved functional vision with a modified prolate intraocular lens. AB - PURPOSE: To evaluate whether the Tecnis Z9000 intraocular lens (IOL) (Pfizer) with a modified prolate anterior surface provides better quality of vision than a conventional spherical IOL. SETTING: Oregon Eye Institute, Eugene, Oregon, USA. METHODS: Patients presenting for cataract surgery who were randomly assigned to receive a Tecnis Z9000 IOL (Pfizer) or a Sensar OptiEdge AR40e IOL (AMO) in 1 eye were followed for 3 months postoperatively. The patient could elect to have the same type of IOL implanted in the fellow eye. The results of sine-wave grating contrast sensitivity testing under mesopic and photopic conditions were compared interindividually. RESULTS: Monocular comparison was made between the 2 IOL groups, which comprised 15 patients each. The Tecnis IOL provided significantly better contrast sensitivity at 6 cycles per degree (cpd) under photopic conditions and at 1.5 and 3 cpd under mesopic conditions. Seven patients with a Tecnis IOL and 9 patients with an AR40e IOL had subsequent implantation in the fellow eye. In all eyes, including fellow eyes, having IOL implantation, the Tecnis provided significantly better contrast sensitivity at 3 and 6 cpd under photopic conditions and at 1.5, 3, and 6 cpd under mesopic conditions. The mean contrast sensitivity in fellow eyes showed that the Tecnis IOL produced significantly better results at some spatial frequencies. CONCLUSIONS: Results show the Tecnis IOL with a modified prolate anterior surface may produce better contrast sensitivity than a standard spherical IOL under mesopic and photopic conditions. Because contrast sensitivity testing correlates well with functional vision, a goal of future research should be to evaluate patient performance using functional tests such as driving simulation. PMID- 15130634 TI - Outcome of second surgery in LASIK cases aborted due to flap complications. AB - PURPOSE: To describe the technique and timing of second refractive surgery after aborted laser in situ keratomileusis (LASIK) due to intraoperative flap complication and determine the final visual outcome. SETTING: Outpatient ambulatory laser vision correction centers. METHODS: This retrospective noncomparative case series included 16 patients (16 eyes) who had a second refractive surgery after initial LASIK surgery was aborted because of a flap complication. Charts were reviewed with attention to initial preoperative data, intraoperative details of the aborted LASIK, postoperative examination, possible causes of the flap complication, timing and technique of second refractive surgery, and final visual outcome. RESULTS: Causes of the aborted LASIK were identified in 13 of 16 eyes (81.2%) and included eye squeezing (5 eyes), loss of suction or machine failure (5 eyes), steep corneas (2 eyes), and learning curve of the surgeon (1 eye). The mean time until the second surgery was 135 days (range 49 to 372 days). Repeat flaps were created deeper and larger than the initially attempted flaps when possible. No patient had a final uncorrected visual acuity (UCVA) worse than 20/30 after the second surgery. Two eyes (12.5%) lost 1 line of best spectacle-corrected visual acuity. CONCLUSION: A planned delayed reoperation after sufficient corneal healing following an intraoperative flap complication can result in satisfactory recovery of UCVA. PMID- 15130635 TI - Using the lens haptic plane concept and thick-lens ray tracing to calculate intraocular lens power. AB - PURPOSE: To develop a methodology for intraocular lens (IOL) power calculation in which the task of predicting the postoperative position of the IOL is separated from the calculation itself. SETTING: Pharmacia, Groningen, The Netherlands. METHODS: The minimum biometry input needed for IOL power calculation is the mean anterior corneal radius and axial length of the eye. The lens haptic plane (LHP) is the plane where the IOL haptics make contact with eye tissue. It is an anatomical site (eg, the equator of the capsular bag) and is independent of the IOL model. The position of the IOL optic in relation to the LHP is determined from the exact design of the IOL. Gullstrand's eye model is adopted to obtain the posterior corneal radius, thickness of the cornea, and refractive indices of the eye media. Thick-lens ray tracing in the paraxial limit is used for the optical calculation. RESULTS: A spreadsheet is given for the calculation. CONCLUSIONS: The methodology developed allows for IOL power calculation from first principles (ie, using true physical distances, radii, and refractive indices as input for the optical calculation). PMID- 15130636 TI - Laser in situ keratomileusis for the correction of residual ametropia after penetrating keratoplasty. AB - PURPOSE: To evaluate the safety, efficacy, and predictability of excimer laser in situ keratomileusis (LASIK) to correct residual myopia and astigmatism after penetrating keratoplasty (PKP). SETTING: Buzard Eye Institute, Las Vegas, Nevada, USA. METHODS: Twenty-six eyes had LASIK at least 1 year after PKP. All eyes were followed for at least 6 months after LASIK; 22 eyes were followed for 12 months. Sutures were removed at a mean of 13 months post PKP. Laser in situ keratomileusis was performed with the Chiron Automated Corneal Shaper microkeratome (Bausch & Lomb) and the Visx Star excimer laser. Before LASIK, the mean spherical equivalent (SE) was -4.94 diopters (D) +/- 2.79 (SD) and the mean astigmatism was 2.71 +/- 2.33 D; all eyes had regular astigmatism or slightly decentered, irregular astigmatism. RESULTS: At the last follow-up, the mean postoperative uncorrected visual acuity (UCVA) was 20/30, the mean SE was -0.35 +/- 0.65 D, and the mean residual astigmatism was 1.06 +/- 0.67 D. Eighty-six percent of patients had an SE within +/-1.00 D of emmetropia and a UCVA of 20/40 or better. Ten eyes (39%) had 1 or more enhancements, which were performed a mean of 6 months after the primary LASIK. Significant complications such as wound dehiscence, epithelial ingrowth, and corneal decompensation did not occur. At the last follow-up, 18% of patients lost 1 line of best corrected visual acuity and 27% gained 1 line. CONCLUSION: Laser in situ keratomileusis appeared to be a reliable and safe procedure to correct residual myopia and astigmatism after PKP. PMID- 15130637 TI - Power modulations in new phacoemulsification technology: improved outcomes. AB - PURPOSE: To evaluate the clinical outcomes of new phacoemulsification technology. SETTING: Oregon Eye Institute, Eugene, Oregon, USA. METHODS: After optimization of surgical parameters, patients were randomly assigned to cataract extraction with 1 of 4 phacoemulsification machines using new technology available within the past 1 to 2 years. Outcomes were compared with previously published results for these same machines obtained before the advent of the new technology. RESULTS: Improvements in effective phaco time, average phaco power, percentage of clear corneas, and 20/40 or better uncorrected visual acuity at the first postoperative visit were noted for most systems using new technology. CONCLUSION: New phacoemulsification technology offers surgical advantages that translate into improved clinical outcomes. PMID- 15130638 TI - Visual field changes after laser in situ keratomileusis in myopic eyes. AB - PURPOSE: To evaluate the effect of acute elevation of intraocular pressure (IOP) during laser in situ keratomileusis (LASIK) on the visual field in myopic eyes. SETTING: Istanbul University, Cerrahpassa Medical Faculty, Istanbul, Turkey. METHODS: Thirty-seven eyes of 37 patients were included in the study. The LASIK procedure was performed using the Hansatome microkeratome (Bausch & Lomb) and the Summit SVS Apex Plus 193 nm argon-fluoride excimer laser. Visual field testing was performed twice before LASIK and 1 day and 24 months after LASIK. Seventy-six points, 6 degrees apart, in the central visual field (Central 30-2) were tested for threshold sensitivity (Humphrey). The mean threshold sensitivity of baseline and post-LASIK visual field examinations was calculated in each patient. Seventy six points of the Central 30-2 test were divided into 21 clusters corresponding to perimetric nerve-fiber bundles derived from Peridata software version 6.2a (Interzaag AG). The mean threshold sensitivity of each cluster and the visual field indices (mean deviation [MD] and corrected pattern standard deviation [CPSD]) in baseline and follow-up visual fields were compared using analysis of variance. RESULTS: The mean age of the 23 men and 14 women was 31.04 years +/- 6.55 (SD). The mean spherical equivalent refraction was -4.21 +/- 1.66 diopters (D) (range -2.25 to -6.75 D). The mean duration of suction was 42.29 +/- 29.06 seconds. The mean visual field sensitivity at baseline and the 2 follow-up examinations was 25.97 +/- 2.04 dB, 25.70 +/- 1.99 dB, and 27.17 +/- 1.68 dB, respectively (P =.181). There was no difference between preoperative and postoperative visual field clusters except in area 13. In area 13, threshold sensitivity was decreased at 1 day (P =.039) and at the preoperative level at 24 months. The MD of the visual fields was -3.53 +/- 1.67 dB, -3.61 +/- 2.91 dB, and -2.61 +/- 1.66 dB at the preoperative and 2 postoperative examinations, respectively (P =.495). The mean CPSD of the visual fields was 1.76 +/- 1.24 dB, 1.42 +/- 0.85 dB, and 1.74 +/- 0.86 dB, respectively (P =.680). CONCLUSION: Laser in situ keratomileusis did not cause visual field defects in mild to moderate myopic patients who had no risk factors that might render the optic nerve more vulnerable to damage. PMID- 15130639 TI - Changes in the shape of the anterior and posterior corneal surfaces caused by mydriasis and miosis: detailed analysis. AB - PURPOSE: To evaluate changes in the anterior and posterior corneal shape, corneal thickness, and anterior chamber depth (ACD) caused by mydriasis or miosis using scanning-slit corneal topography. SETTING: Department of Ophthalmology, Iwate Medical University School of Medicine, Iwate, Morioka, Japan. METHODS: Twenty eight eyes of 28 healthy volunteers with refractive errors of -6.00 to +0.25 diopters were studied. One eye of each subject had instillation of tropicamide phenylephrine hydrochloride (Mydrin P) to obtain mydriasis and of pilocarpine hydrochloride 2% (Sanpilo) to obtain miosis. To assess the corneal shape, the best-fit sphere (BFS), axial power, and tangential power were measured for the anterior and posterior corneal surfaces before and after mydriasis and before and after miosis using scanning-slit corneal topography (Orbscan version 3.0, Orbtek, Inc.). The pupil size, corneal thickness, and ACD were also examined before and after mydriasis and before and after miosis. RESULTS: The mean age of the patients was 31.1 years +/- 5.6 (SD) (range 20 to 46 years). The anterior BFS changed from a mean of 8.04 +/- 0.3 mm at the time of mydriasis to a mean of 8.00 +/- 0.3 mm at the time of miosis. The posterior BFS changed from 6.53 +/- 0.3 mm to 6.46 +/- 0.3 mm, respectively. Thus, the anterior and posterior cornea became significantly steeper after miosis (P<.01). The ACD was significantly more shallow after miosis than after mydriasis. However, there was no significant difference in corneal thickness after mydriasis or miosis. CONCLUSIONS: The anterior and posterior corneal shapes changed as a result of mydriasis and miosis, and the refractive power of the cornea significantly increased after miosis. To date, changes in refractive power from changes in pupil size have been attributed to a change in the refractive power of the crystalline lens; however, it is now thought that changes in corneal refractive power also occur. PMID- 15130640 TI - Reproducibility of LASIK flap thickness using the Hansatome microkeratome. AB - PURPOSE: To evaluate the actual versus the expected thickness of laser in situ keratomileusis (LASIK) flaps and to determine the factors that affect flap thickness. SETTING: Centre For Sight, Queen Victoria Hospital, East Grinstead, United Kingdom. METHODS: A retrospective analysis of LASIK procedures in 757 consecutive eyes was done. The surgery was performed by a single surgeon using 2 Hansatome microkeratomes (Bausch & Lomb) with 160 microm and 180 microm heads. Patient age, preoperative manifest refraction, automated keratometry, preoperative central pachymetry, and intraoperative stromal pachymetry were evaluated to determine whether they influenced the actual flap thickness. RESULTS: Bilateral LASIK was performed in 343 patients (686 eyes). The 160 microm head was used in 641 eyes (84.6%) (Group 1) and the 180 microm head, in 116 eyes (15.4%) (Group 2). The mean preoperative manifest refraction spherical equivalent (SE) was -3.9 diopters (D) +/- 4.5 (SD) (range +7.4 to -25.0 D) in Group 1 and 4.4 +/- 3.7 D (range +7.1 to -12.9 D) in Group 2. The mean preoperative keratometry reading was 43.6 +/- 1.8 D (range 36.0 to 48.6 D) and 43.6 +/- 1.8 D (range 35.9 to 47.0 D), respectively; the mean preoperative central pachymetry was 543 +/- 35 microm (range 447 to 643 microm) and 548 +/- 31 microm (range 453 to 613 microm), respectively; and the mean flap thickness was 116.4 +/- 19.8 microm and 117.3 +/- 18.0 microm, respectively. The difference between the actual and the expected flap thickness in each group was statistically significant (P<.001). There was no significant difference in the actual flap thickness between the 2 Hansatomes. The preoperative SE and central pachymetry were the only factors that influenced the actual flap thickness (P<.05); thin flaps were more common with increasing myopia and increasing corneal thickness. CONCLUSIONS: Spherical equivalent and preoperative pachymetry were the principal factors that influenced flap thickness. Preoperative keratometry values and patient age did not influence the actual flap thickness. PMID- 15130641 TI - Peripheral corneal relaxing incisions after excimer laser refractive surgery. AB - PURPOSE: To evaluate the effectiveness of peripheral corneal relaxing incisions (PCRIs) for correcting corneal astigmatism after excimer laser refractive surgery. SETTING: Cullen Eye Institute, Baylor College of Medicine, Houston, Texas, USA. METHODS: In this retrospective case series, PCRIs were performed in 33 eyes (30 patients) that had residual astigmatism after photorefractive keratectomy or laser in situ keratomileusis according to a nomogram based on age and preoperative refractive astigmatism. Uncorrected visual acuity (UCVA) and refractive and keratometric astigmatism were evaluated, and vector analysis using the Holladay-Cravy-Koch formula was performed. RESULTS: The percentage of eyes with a UCVA of 20/20 or better increased significantly from 6% (2/33) preoperatively to 61% (20/33) postoperatively (P<.001). Refractive astigmatism was reduced significantly, and the effect was stable up to 1 year after PCRIs. The percentage of eyes within +/-0.5 diopter (D) and +/-1.0 D of cylinder increased by 73% and 52%, respectively (both P<.001). No eye lost 1 or more lines of best spectacle-corrected visual acuity. CONCLUSION: Peripheral corneal relaxing incisions are an effective approach for correcting low amounts of corneal astigmatism in eyes that have had excimer laser refractive surgery. PMID- 15130642 TI - Axial length and age at cataract surgery. AB - PURPOSE: To examine the relationship between the axial length of the eye and age at surgery for cataract. SETTING: Postgraduate teaching hospital. METHODS: A retrospective review of a consecutive series of 1776 patients older than 40 years was performed. The patient's age at first-eye surgery, axial length, and preoperative and postoperative best corrected visual acuities were recorded. Linear regression analysis was performed using the age at time of cataract surgery as the dependent variable. RESULTS: Data were available for 1576 of 1670 eyes eligible for inclusion. There was strong evidence of a weak association between a decrease in age at surgery with an increase in axial length (P<.001). The mean patient age at surgery was 74 years for eyes with an axial length less than 25.0 mm, 67 years for eyes with an axial length between 25.0 mm and 30.0 mm, and 61 years for eyes with axial length greater than 30.0 mm. CONCLUSION: An increase in the axial length of the eye was associated with a lower mean age at time of cataract surgery. PMID- 15130643 TI - Scleral fixation of dislocated posterior chamber intraocular lenses: Temporary haptic externalization through a clear corneal incision. AB - PURPOSE: To report the results of managing dislocated posterior chamber intraocular lenses (PC IOLs) by externalizing the haptics through a clear corneal incision. SETTING: The Retina Center at Pali Momi, Aiea, Hawaii, USA. METHODS: This retrospective consecutive series comprised cases in which a dislocated PC IOL was managed with pars plana vitrectomy. With this method, the dislocated PC IOL is retrieved and stabilized in the anterior chamber. The haptic is externalized through a clear corneal incision for suture knot placement and then reinserted. A scleral fixation suture is placed 1.25 mm posterior to the limbus under a scleral flap. RESULTS: The study included 14 eyes of 14 patients with a minimum follow-up of 6 months (median 15.4 months). There were 2 subluxated PC IOLs, 11 posteriorly dislocated PC IOLs, and 1 in-the-bag IOL dislocation. Postoperative vision and/or visual symptoms were stable or improved in 86% of eyes. Two eyes had worse vision caused by conditions not related to surgery including chronic cystoid macular edema from latanoprost use and optic atrophy. All IOLs were well fixated and stable, although 1 eye with asymmetric haptics had a slightly tilted IOL. CONCLUSIONS: Dislocated PC IOL management by externalizing the haptic through a clear corneal incision stabilized the IOL in the anterior chamber, minimized intraocular operative manipulations, and allowed easier placement of the opposite haptic over residual capsule, if available. Stable fixation was achieved without dislocation or IOL-related complications recurring. PMID- 15130644 TI - Optimization of contact lens fitting in keratectasia patients after laser in situ keratomileusis. AB - PURPOSE: To evaluate the clinical outcomes of contact lens fitting in eyes with keratectasia after laser in situ keratomileusis (LASIK) to optimize the performance of the contact lenses. SETTING: Contact Lens Clinic, Seoul National University Hospital, Seoul, Korea. METHODS: Six eyes of 4 patients with keratectasia were examined. Keratectasia was diagnosed based on abnormal topographic findings, progressive myopic changes, and uncorrected visual acuity with glasses. A multicurve lens or reverse-geometry lens (RGL) was selected based on the anterior elevation map. The base curve radius and peripheral parameters were determined based on the axial map and the evaluation of the fluorescein pattern of the lens. The contact lens fitting characteristics and visual acuity were evaluated. Patients were interviewed about their level of comfort, daily contact lens wear time, and glare. RESULTS: Multicurve lenses and RGLs were fitted in 4 eyes and 2 eyes, respectively. The base curve radius of the multicurve lenses ranged from 7.2 to 7.5 mm; the alignment curve radius of RGLs was 7.6 mm in both cases. The radii of peripheral curves were customized to enhance tear interchange. Visual acuity improved to 20/30 or better in all eyes. All patients were completely satisfied with the comfort of the fitting. The mean daily wearing time was 12.7 hours. One patient reported tolerable glare in the eye after being fitted with an RGL. CONCLUSION: Proper contact lens fitting based on topographic data and slitlamp evaluation allowed good visual rehabilitation and comfortable extended daily wear in patients with keratectasia after LASIK. PMID- 15130645 TI - Residual bed thickness and corneal forward shift after laser in situ keratomileusis. AB - PURPOSE: To prospectively assess the forward shift of the cornea after laser in situ keratomileusis (LASIK) in relation to the residual corneal bed thickness. SETTING: Miyata Eye Hospital, Miyazaki, Japan. METHODS: Laser in situ keratomileusis was performed in 164 eyes of 85 patients with a mean myopic refractive error of -5.6 diopters (D) +/- 2.8 (SD) (range -1.25 to -14.5 D). Corneal topography of the posterior corneal surface was obtained using a scanning slit topography system before and 1 month after surgery. Similar measurements were performed in 20 eyes of 10 normal subjects at an interval of 1 month. The amount of anteroposterior movement of the posterior corneal surface was determined. Multiple regression analysis was used to assess the factors that affected the forward shift of the corneal back surface. RESULTS: The mean residual corneal bed thickness after laser ablation was 388.0 +/- 35.9 microm (range 308 to 489 microm). After surgery, the posterior corneal surface showed a mean forward shift of 46.4 +/- 27.9 microm, which was significantly larger than the absolute difference of 2 measurements obtained in normal subjects, 2.6 +/- 5.7 microm (P<.0001, Student t test). Variables relevant to the forward shift of the corneal posterior surface were, in order of magnitude of influence, the amount of laser ablation (partial regression coefficient B = 0.736, P<.0001) and the preoperative corneal thickness (B = -0.198, P<.0001). The residual corneal bed thickness was not relevant to the forward shift of the cornea. CONCLUSIONS: Even if a residual corneal bed of 300 microm or thicker is preserved, anterior bulging of the cornea after LASIK can occur. Eyes with thin corneas and high myopia requiring greater laser ablation are more predisposed to an anterior shift of the cornea. PMID- 15130647 TI - Functional outcomes of acrylic intraocular lenses in pediatric cataract surgery. AB - PURPOSE: To evaluate the functional outcomes of in-the-bag implantation of acrylic intraocular lenses (IOLs) with posterior continuous curvilinear capsulorhexis (PCCC), without PCCC, with PCCC and anterior vitrectomy, and with PCCC and optic capture in pediatric cataract surgery. SETTING: Pediatric Ophthalmology Service, Guru Nanak Eye Centre, New Delhi, India. METHODS: Forty two eyes of 25 children were included in this prospective study. All eyes had in the-bag implantation of an AcrySof IOL (Alcon). Twenty-five eyes had had an anterior continuous curvilinear capsulorhexis (ACCC) (Group A). Seventeen eyes had PCCC along with ACCC (Group B), 4 had anterior vitrectomy combined with PCCC (Group C), and 6 had PCCC with IOL optic capture through the PCCC (Group D). Secondary opacification of the visual axis, visual acuity, and possible complications were observed and analyzed. RESULTS: The mean age of the patients was 78 months (range 36 to 144 months). The mean follow-up was 13 months (range 6 to 18 months). Four eyes (16%) in Group A developed visually significant posterior capsule opacification (PCO) involving the central visual axis and required secondary capsulotomy. All eyes in Groups B, C, and D had a clear visual axis at the last follow-up and did not require a secondary procedure. Minimal postoperative inflammation (ie, aqueous flare and IOL deposits ) was seen in all groups. The mean preoperative decimal best corrected visual acuity (BCVA) in Groups A, B, C, and D was 0.095, 0.055, 0.174, and 0.039, respectively. Postoperatively, the BCVA was 0.54, 0.66, 0.66, and 0.66, respectively. CONCLUSIONS: An optimal-sized ACCC followed by in-the-bag implantation of a foldable acrylic IOL helped maintain a clear visual axis by delaying the onset of PCO and leading to milder PCO. The benefits of a foldable acrylic IOL in pediatric cataract surgery can be increased by combining it with PCCC, with or without anterior vitrectomy, or with optic capture of the IOL. PMID- 15130646 TI - Visual axis opacification after AcrySof intraocular lens implantation in children. AB - PURPOSE: To evaluate visual axis opacification after AcrySof intraocular lens (IOL) (Alcon) implantation in pediatric eyes. SETTING: Iladevi Cataract and IOL Research Centre, Ahmedabad, India. METHODS: This prospective study evaluated 103 consecutive eyes of 72 children with congenital cataract. Two groups were formed based on age at surgery: Group 1, younger than 2 years, and Group 2, older than 2 years. All eyes in Group 1 (n = 37) had primary posterior continuous curvilinear capsulorhexis (PCCC) with anterior vitrectomy. In Group 2 (n = 66), management of the posterior capsule was assigned randomly to no PCCC (Group 2A, n = 37) or PCCC (Group 2B, n = 29). The PCCC group was further randomized into 2 subgroups: no vitrectomy (Group 2BN, n = 14) or vitrectomy (Group 2BV, n = 15). The primary outcome measures were visual axis opacification and the resulting need for a secondary procedure. Statistical analysis was performed using SPSS for Windows (version 11.0.1). RESULTS: The mean age of the patients was 5.2 years +/- 5.0 (SD) (range 0.2 to 16.0 years) and the mean follow-up, 2.3 +/- 0.9 years (range 1.0 to 4.0 years). Overall, 41 eyes (39.8%) developed visual axis opacification and 14 (13.6%) required secondary intervention. In Group 1, 4 eyes (10.8%) developed visual axis opacification and 3 (8.1%) had a secondary pars plana vitrectomy. In Group 2A, 31 eyes (83.8%) developed posterior capsule opacification (PCO) and 10 eyes (27.7%) had secondary intervention. Children 8 years or younger at the time of surgery developed significantly greater PCO than older children (P =.01). Five eyes (37.5%) in Group 2BN had opacification of the anterior vitreous face, 1 of which required a secondary procedure. One eye (6.7%) in Group 2BV had visual axis opacification that did not require a secondary procedure. CONCLUSIONS: AcrySof IOL implantation with appropriate management of the posterior capsule maintained a clear visual axis in 60.2% of eyes. Of the 39.8% of eyes with visual axis opacification, 13.6% had visually significant opacification and required a secondary procedure. PMID- 15130648 TI - Foldable versus rigid intraocular lenses in conjunction with pars plana vitrectomy and other vitreoretinal procedures. AB - PURPOSE: To compare the results of foldable acrylic intraocular lens (IOL) implantation through a clear corneal incision with those of rigid IOL implantation in eyes having pars plana vitrectomy (PPV). SETTING: Tertiary referral-based university institute. METHODS: A consecutive retrospective comparative chart review was performed in all eyes that had PPV and foldable IOL implantation between May 15, 1999, and November 1, 2000 (n = 30), and all eyes that had PPV and rigid IOL implantation between April 1, 1996, and May 14, 1999 (n = 30). Preoperative baseline data and postoperative outcome data were recorded. Pars plana vitrectomy and associated vitreoretinal procedures were performed as indicated according to individual circumstances. A minimum of 1 week of follow-up information was available for all eyes. RESULTS: Baseline characteristics in both groups of patients, including age, sex, eye involved, and phakic state, were similar. The preoperative visual acuities were also similar, ranging from 20/30 to hand motions; the mean visual acuity was 20/200. The IOL was implanted in all eyes uneventfully and did not restrict fundoscopy. The mean follow-up was significantly longer in the rigid IOL group (20 months) than in the foldable IOL group (7 months) (P<.001), probably because of the earlier case acquisition. The mean postoperative best corrected visual acuity was 20/200 in the foldable IOL group and 20/100 in the rigid IOL group. There was no difference between the 2 groups in the rate of postoperative retinal detachment, recurrent macular hole, or repeat PPV. Elevated intraocular pressure (IOP) on the first postoperative day was more common in the rigid IOL group than in the foldable IOL group (P =.078) because more patients in the rigid IOL group had surgery for diabetic ocular complications and these patients had a greater IOP rise. CONCLUSION: Acrylic IOLs can be safely implanted in conjunction with PPV in selected cases. PMID- 15130649 TI - Anterior chamber angle biometry with ultrasound biomicroscopy after diode laser thermal keratoplasty. AB - PURPOSE: To evaluate the effect of diode laser thermal keratoplasty (DTK) on the geometry of the anterior chamber angle with ultrasound biomicroscopy (UBM). SETTING: Department of Ophthalmology and Optometry, Paracelsus Private Medical University, Salzburg, Austria. METHODS: In 9 eyes, a Rodenstock DTK (ProLaser Medical Systems) was used to correct hyperopia (n = 6) or hyperopic astigmatism (n = 3). Hyperopia was treated with a single- or double-ring pattern and hyperopic astigmatism, with additional spots in each of the flat half meridians. Ultrasound biomicroscopy measurements of the chamber angle were obtained preoperatively and 3 months postoperatively using a Humphrey UBM 840 system with a 50 MHz transducer. Quantitative measurements were performed at 3-, 6-, 9-, and 12-o'clock positions using a standardized method. RESULTS: No statistically significant differences between the preoperative and postoperative chamber angle measurements were detected. CONCLUSIONS: The results of the UMB measurements were surprising given the flattening of the corneal periphery found by computerized topography, a finding that suggests narrowing of the chamber angle. Shortening of the corneal tissue may result in centripetal traction that affects the trabecular meshwork and results in a relative opening of the chamber angle structures. PMID- 15130650 TI - Water-mediated lysis of lens epithelial cells attached to lens capsule. AB - PURPOSE: To investigate the effect of distilled deionized water (DDW) on lens epithelial cells (LECs) attached to the lens capsule. SETTING: Wound Healing Research Laboratory, Center for Vision Research, Westmead Hospital, Sydney, NSW, Australia. METHODS: Anterior capsulotomy specimens taken during routine cataract surgery were divided in half. One half was immersed in DDW and the other half in culture medium (control) for 1 to 5 minutes and photographed at intervals by phase-contrast microscopy. In further experiments, the capsules were exposed to DDW for 1 or 2 minutes and placed in culture for 1 week to determine whether LECs survive treatment and are capable of repopulating the lens capsule. RESULTS: Distilled-deionized water induced marked swelling of the cytoplasm within 60 seconds of treatment. At 120 seconds, there was disruption of the plasma membranes, with few intact cells remaining. In the control capsules, confluent monolayers of LECs covered the entire capsule surface with a halo of LECs growing on the surrounding plastic well. Viable LECs were observed in 1 of 3 capsules treated for 1 minute with DDW. These did not reach confluence or grow off the capsule onto the surrounding well. No viable LECs were seen on capsules exposed to DDW for 2 minutes. CONCLUSIONS: Short exposure of LECs to DDW induced extensive and rapid cell lysis. Distilled-deonized water may be a useful agent for instillation in the capsular bag during sealed-capsule irrigation to prevent posterior capsule opacification. PMID- 15130651 TI - Femtosecond laser LASIK flap preparation with conical incision and positional spikes. AB - PURPOSE: To describe the creation of corneal flaps for laser in situ keratomileusis (LASIK) with conical incisions and positional spikes that may increase postoperative rotational flap stability. SETTING: University Eye Department. METHODS: A corneal flap was prepared in 5 freshly enucleated porcine eyes using a femtosecond laser. In the first step, the flap bed with a diameter of 8.0 mm was created by an intrastromal incision running parallel to the corneal surface. In the second step, a posterior-anterior incision was created with a diameter of 8.0 mm at the level of the first incision and 7.0 mm at the corneal surface. In addition, a hinge and 3 spikes were prepared in the flap with corresponding notches in the surrounding tissue. RESULTS: In all eyes, the corneal flaps were prepared easily within 5 minutes. CONCLUSION: Femtosecond laser technology allowed a new type of LASIK flap formation with conical incisions and positional spikes. PMID- 15130652 TI - Legacy AdvanTec and Sovereign WhiteStar: a wound temperature study. AB - PURPOSE: To assess the wound temperature of the Sovereign WhiteStar (S-WS) (AMO) and Legacy AdvanTec (L-ADV) (Alcon) phacoemulsification systems. SETTING: John A. Moran Eye Center, Health Sciences Center, University of Utah, Salt Lake City, Utah, USA. METHODS: Phacoemulsification using 20-gauge, 30-degree straight tips with the L-ADV and S-WS systems was performed in fresh cadaver eyes. The power was set at 50%, and aspiration was 12 mL/min; the L-ADV was run at 15 pulses per second (pps) and the S-WS at WS CF (6 milliseconds on, 12 milliseconds off). Temperature was measured at 5-second intervals for 60 seconds using a microthermistor placed in the wound. The phaco tip was angled 30 degrees to increase wound-tissue contact. At 10 seconds, the flow was clamped to simulate occlusion. Five runs, sleeved and unsleeved, were averaged for the phaco tip. RESULTS: The mean temperature was significantly higher with L-ADV than with S-WS (from 10 seconds on in the sleeved condition and from 5 seconds on in the unsleeved condition after the aspiration line was clamped). In 2 of 5 sleeved runs and 4 of 5 unsleeved runs, the L-ADV handpiece decreased power as the temperature increased. One run with the L-ADV with sleeve showed signs of wound burn. There were no signs of wound burn with the S-WS. The highest temperature recorded was 57.5 degrees C with the L-ADV and 38.6 degrees C with the S-WS. Power tests showed L-ADV protected the stroke length and S-WS had constant power except in air where it increased power. CONCLUSIONS: There was less increase in wound temperature over time with the S-WS than with the L-ADV system in sleeved and unsleeved simulated surgery in human eye-bank eyes. Because the ultrasound handpieces respond differently under different load scenarios, meaningful comparisons of pulsing features are difficult to create. PMID- 15130653 TI - Capsular bag opacification after experimental implantation of a new accommodating intraocular lens in rabbit eyes. AB - PURPOSE: To evaluate the development of capsular bag opacification in rabbit eyes after implantation of an intraocular lens (IOL) designed to minimize contact between the anterior capsule and the IOL and ensure expansion of the capsular bag. SETTING: David J. Apple, MD Laboratories for Ophthalmic Devices Research, John A. Moran Eye Center, University of Utah, Salt Lake City, Utah, USA. METHODS: Ten New Zealand white rabbits had a study IOL (new accommodating silicone IOL [Synchrony, Visiogen, Inc.]) implanted in 1 eye and a control IOL (1-piece plate silicone IOL with large fixation holes) implanted in the other eye. Intraocular lens position, anterior capsule opacification (ACO), and posterior capsule opacification (PCO) were qualitatively assessed using slitlamp retroillumination photographs of the dilated eyes. Anterior capsule opacification and PCO were graded on a 0 to 4 scale after the eyes were enucleated (Miyake-Apple posterior and anterior views after excision of the cornea and iris). The eyes were also evaluated histopathologically. RESULTS: The rate of ACO and PCO was significantly higher in the control group. Fibrosis and ACO were almost absent in the study group; the control group exhibited extensive capsulorhexis contraction, including capsulorhexis occlusion. Postoperative IOL dislocation into the anterior chamber and pupillary block syndrome were observed in some eyes in the study group. CONCLUSIONS: The special design features associated with the study IOL appeared to help prevent PCO. Complications in the study group were probably caused by the increased posterior vitreous pressure in rabbit eyes compared to human eyes and the relatively large size of the study IOL relative to the anterior segment of rabbit eyes. PMID- 15130654 TI - Late corneal scarring after retinal detachment surgery 42 months after photorefractive keratectomy. AB - A 42-year-old man had uneventful bilateral nonsimultaneous photorefractive keratectomy (PRK) for severe myopia. Thirty-nine months after the procedure, the patient presented with a retinal detachment (RD) in the right eye. Cerclage, vitrectomy, endolaser, and intravitreal silicone oil tamponade were performed, and the RD was successfully repaired. Three months after vitrectomy and 42 months after PRK, the patient complained of visual impairment in the right eye and photophobia. On slitlamp examination, marked reticular scarring of the central anterior cornea was observed. The occurrence of late-onset corneal haze highlights the need for special attention to patients who have vitrectomy after PRK. PMID- 15130655 TI - Correction of spherical myopia with a single 150-degree intrastromal corneal ring segment. AB - A 47-year-old woman with an uncorrected visual acuity (UCVA) of 20/70 and a refraction of -1.50 +0.75 x 75 in the right eye had 2 0.25 mm intrastromal corneal ring segments (Intacs) inserted uneventfully through a superior incision. The patient was hyperopic 2 years later with a refraction of +0.50 +0.75 x 25 and was intolerant of spectacles. She also complained of temporal glare. Twenty-seven months after insertion, the temporal ring segment was removed. Four months later, the UCVA was 20/20; with a refraction of plano +0.50 x 35, the visual acuity was 20/15. Topography showed corresponding regular astigmatism, and the patient's glare had resolved. Removal of 1 Intacs segment may be an option in cases of overcorrection after ring insertion for myopia. PMID- 15130656 TI - Capsular block syndrome caused by a reversed-optic intraocular lens. AB - A 52-year-old man who had phacoemulsification and in-the-bag fixation of a posterior chamber intraocular lens (IOL) developed capsular block syndrome 1 day after reverse implantation of the IOL. After a neodymium:YAG (Nd:YAG) laser peripheral anterior capsulotomy, the distended capsular bag collapsed significantly and the artificial myopia partially resolved. An Nd:YAG laser peripheral anterior capsulotomy appeared to be an effective, simple, and minimally invasive method to treat this problem. PMID- 15130657 TI - Long-term observation of the refraction with a reversed-optic posterior chamber intraocular lens. AB - A 73-year-old man had phacoemulsification and implantation of a posterior chamber intraocular lens (PC IOL) in both eyes. In the right eye, the optic was unintentionally reversed in the bag. In the left eye, the posterior capsule was ruptured and the IOL haptics were fixated in the ciliary sulcus. The refraction in the sulcus-fixated left eye was stable from 1 month to 4 years. The right eye had a myopic shift at 1 month, with a regression of 1.50 diopters that continued for 2 years. The final IOL position in the right eye was 0.25 mm anterior to the position predicted by the SRK/T formula. It took 2 years for the refraction in the eye with the reversed-optic PC IOL to become stable. A large myopic shift and gradual regression in refraction may stem from a different wound-healing reaction than that of properly fixated IOLs. PMID- 15130658 TI - Refractive lensectomy and cross-cylinder laser in situ keratomileusis for the correction of extreme hyperopic astigmatism. AB - Refractive lensectomy followed by cross-cylinder laser in situ keratomileusis was performed in both eyes of a 46-year-old patient with extreme hyperopic astigmatism. Six months postoperatively, the uncorrected visual acuity was 20/25 with a manifest refraction of +0.25 -0.50 x 44 in the right eye and +0.25 -0.25 x 10 in the left eye. The best corrected visual acuity remained unchanged in both eyes at 20/25. Refractive lensectomy and cross-cylinder LASIK can be effective for treating extreme hyperopic astigmatism. PMID- 15130659 TI - Spontaneous disinsertion of a multipiece foldable acrylic intraocular lens haptic 3 and 12 months after implantation. AB - We report 2 cases of spontaneous postoperative haptic disinsertion with the Alcon AcrySof MA60BM posterior chamber intraocular lens (PC IOL). A 14-year-old girl with a history of acute retinal necrosis and pars plana vitrectomy with lensectomy for retinal detachment repair had secondary implantation of a PC IOL in the ciliary sulcus. Three months later, the superior haptic disinserted from the optic and dislocated into the anterior chamber. In the second case, a 22-year old man had lens aspiration and PC IOL implantation in the capsular bag. One year after surgery, the PC IOL had a similar problem, with the disinserted superior haptic remaining in the capsular bag while the optic and inferior haptic dislocated into the anterior chamber. In both patients, the PC IOLs were exchanged for rigid single-piece PC IOLs. The patients have had no further problems. PMID- 15130660 TI - Surgical management of early postoperative capsular bag distension syndrome. PMID- 15130661 TI - Management of radiotherapy-induced cataracts in eyes with retinoblastoma. PMID- 15130662 TI - Epidemiology in aging research. AB - Epidemiology and Gerontology are relatively young scientific disciplines, both of which have had their own specific development in the past few decades. Only very recently, expertise from both fields has been brought together to extend and strengthen the spectrum of aging research. In this review, we outline the definition and methods of epidemiology and its historical development as a scientific discipline. We review and give examples of past and current contributions of epidemiology to aging research, and we outline the specific challenges, difficulties and opportunities encountered by epidemiologists in this field. We end up with an outlook on major challenges and opportunities for further development of epidemiology in aging research. We conclude that further integrative approaches with a particular emphasis on functional impairments are needed. Furthermore, the etiology of chronic diseases and impairments among the elderly cannot be studied by restricting the focus to health outcomes and their determinants at old age. Major efforts have to be made to incorporate measures of relevant determinants throughout the life span. Finally, epidemiologic methods should be more widely used for studies of medical and nursing care, and results of epidemiologic research need to be more effectively transferred into the practice of prevention. PMID- 15130663 TI - Inflammatory mediators in the elderly. AB - Ageing is accompanied by 2-4-fold increases in plasma/serum levels of inflammatory mediators such as cytokines and acute phase proteins. A wide range of factors seems to contribute to this low-grade inflammation, including an increased amount of fat tissue, decreased production of sex steroids, smoking, subclinical infections (e.g. asymptomatic bacteriuria), and chronic disorders such as cardiovascular diseases and Alzheimer's disease. Furthermore, there is some evidence that ageing is associated with a dysregulated cytokine response following stimulation. Several inflammatory mediators such as tumour necrosis factor-alpha and interleukin-6 have the potential to induce/aggravate risk factors in age-associated pathology, providing a positive feedback mechanism. Thus, it is possible that inflammatory mediators constitute a link between life style factors, infections and physiological changes in the process of ageing on the one hand and risk factors for age-associated diseases on the other. Consistent with this, inflammatory mediators are strong predictors of mortality independently of other known risk factors and co-morbidity in elderly cohorts. A direct pathogenetic role of inflammatory mediators would be highly likely if longevity was shown to be associated with cytokine polymorphisms regulating cytokine production. Several studies support indeed this hypothesis but, unfortunately, findings in this area are conflicting, which probably reflects the complexity of the effect of cytokine polymorphisms and their interaction with the lifestyle and sex. PMID- 15130664 TI - Myocyte aging and mitochondrial turnover. AB - Cardiac myocytes, skeletal muscle fibers, and other long-lived postmitotic cells show dramatic age-related alterations that mainly affect mitochondria and the lysosomal compartment. Mitochondria are primary sites of reactive oxygen species formation that causes progressive damage to mitochondrial DNA and proteins in parallel to intralysosomal lipofuscin accumulation. There is amassing evidence that several various mechanisms may contribute to age-related accumulation of damaged mitochondria following initial oxidative injury. Such mechanisms may include clonal expansion of defective mitochondria, decreased propensity of altered mitochondria to become autophagocytosed (due to mitochondrial enlargement or decreased membrane damage associated with weakened respiration), suppressed autophagy because of heavy lipofuscin loading of lysosomes, and decreased efficiency of Lon protease. PMID- 15130665 TI - Expression of the human ferritin light chain in a frataxin mutant yeast affects ageing and cell death. AB - Ferritin is one of the major eukaryotic proteins involved in regulating iron metabolism and maintaining iron homeostasis. However, Saccaromyces cerevisiae is an exception, possessing no ferritin and using other means to store excess iron. The only potential iron storage protein identified in yeast so far is the homologue of human frataxin (YFH1p). In this study, we found that dysfunction of yeast frataxin shortens mean lifespan by 49% compared to the WT control. Interestingly, the human ferritin L gene can, at least partially, complement the function of yeast frataxin, extending lifespan and protecting cells from death induced by oxidative stress or excess iron. Our findings indicate that ferritin L can perform functions in yeast that are similar to its functions in mammals, and suggest that common mechanisms may exist for preventing iron and oxidative damage in single- and multi-cellular eukaryotic organisms. Clearly, elucidation of the function of human ferritin in yeast would help in gaining a better understanding the molecular basis of iron storage diseases. PMID- 15130666 TI - Aging of Hutchinson-Gilford progeria syndrome fibroblasts is characterised by hyperproliferation and increased apoptosis. AB - Hutchinson-Gilford progeria syndrome is a rare genetic disorder that mimics certain aspects of aging prematurely. Recent work has revealed that mutations in the lamin A gene are a cause of the disease. We show here that cellular aging of Hutchinson-Gilford progeria syndrome fibroblasts is characterised by a period of hyperproliferation and terminates with a large increase in the rate of apoptosis. The occurrence of cells with abnormal nuclear morphology reported by others is shown to be a result of cell division since the fraction of these abnormalities increases with cellular age. Similarly, the proportion of cells with an abnormal or absent A-type lamina increases with age. These data provide clues as to the cellular basis for premature aging in HGPS and support the view that cellular senescence and tissue homeostasis are important factors in the normal aging process. PMID- 15130667 TI - Modification of the longevity-related degree of fatty acid unsaturation modulates oxidative damage to proteins and mitochondrial DNA in liver and brain. AB - Previous studies have shown that tissue fatty acid unsaturation correlates inversely with maximum longevity. However, it is unclear if this is related to the effects of fatty acid unsaturation only on lipids, or also on proteins and DNA, specially on mitochondrial DNA (mtDNA) oxidative damage. In this investigation the degree of fatty acid unsaturation of liver and brain was successfully manipulated in Wistar rats by chronic feeding with specially designed semipurified diets rich in saturated or unsaturated fats. The brain, an organ of special relevance for aging, was most profoundly affected by the increase in fatty acid unsaturation, and showed significant increases in malondialdehyde (MDA)-lysine, aminoadipic semialdehyde (a protein carbonyl), N(epsilon)-(carboxymethyl)lysine, and N(epsilon)-(carboxyethyl)lysine in proteins, as well as in 8-oxo,7,8-dihydro-2'-deoxyguanosine (8-oxodG) in mtDNA without changes in nuclear DNA (nDNA). In the liver 8-oxodG was also increased in mtDNA and not in nDNA. These DNA results are consistent with the presence of a high density of mitochondrial inner membranes (rich in lipids and in reactive oxygen species generation capacity) near mtDNA but not near nDNA. Among the protein markers analyzed, MDA-lysine was most consistent and responsive to fatty acid unsaturation, since it increased in both organs and showed the highest increase. These results, together with previous data from our laboratories, show that increasing the degree of fatty unsaturation of postmitotic tissues in vivo can raise not only lipid but also protein and mtDNA oxidative damage. This is mechanistically relevant in relation to the constitutively low tissue fatty acid unsaturation of long-lived animals. PMID- 15130668 TI - Elevated gadd153/chop expression and enhanced c-Jun N-terminal protein kinase activation sensitizes aged cells to ER stress. AB - The endoplasmic reticulum (ER), as a processing plant for the folding and posttranslational modification of proteins, is exquisitely sensitive to changes in its internal environment. Various conditions, collectively termed 'ER stress', can perturb ER functions, leading to the activation of a complex response known as the unfolded protein response. Here, we investigated the response of hepatocytes derived from young (4-5 months) and aged (24-26 months) rats to two agents, thapsigargin (TG) and tunicamycin (TM), which act via different mechanisms to induce ER stress. Old hepatocytes displayed greater cell death than young cells following treatment with TG or TM, associated with higher expression of the pro-apoptotic gene gadd153 (also known as chop) and enhanced c-Jun N terminal protein kinase (JNK) activation. Pharmacologic inhibition of JNK decreased the expression of TG-stimulated gadd153 in old cells and reduced their sensitivity to TG-induced cell death. Inhibition of p38, on the other hand, enhanced TG-induced gadd153 expression and JNK activation, and augmented TG induced cell death. Additional experiments implicated the PERK/eIF-2 alpha signaling pathway as a contributor to the higher Gadd153 expression and JNK activation, and greater sensitivity of old cells to ER stress. PMID- 15130669 TI - Mitochondrial and sarcoplasmic proteins, but not myosin heavy chain, are sensitive to leucine supplementation in old rat skeletal muscle. AB - Leucine has a major anabolic impact on muscle protein synthesis in young as in old animals. However, myosin heavy chain (MHC), sarcoplasmic and mitochondrial proteins may differently respond to anabolic factors, especially during aging. To test this hypothesis, fractional synthesis rates (FSR) of the three muscle protein fractions were measured using a flooding dose of [1-(13)C] phenylalanine, in gastrocnemius muscle of adult (8 months) and old (22 months) rats, either in postabsorptive state (PA), or 90-120 min after ingestion of a alanine supplemented meal (PP+A) or a leucine-supplemented meal (PP+L). In adult and old rats, in comparison with PA, leucine stimulated mitochondrial (adult: 0.260+/ 0.011 vs 0.238+/-0.012%h(-1); old: 0.289+/-0.010 vs 0.250+/-0.010%h(-1); PP+L vs PA, P<0.05) and sarcoplasmic (adult: 0.182+/-0.011 vs 0.143+/-0.006%h(-1); old: 0.195+/-0.010 vs 0.149+/-0.008%h(-1); PP+L vs PA, P<0.05) protein FSR, but not MHC synthesis in old rats (0.101+/-0.009 vs 0.137+/-0.018%h(-1); PP+L vs PA, P=NS). In conclusion, synthesis of specific muscle protein is activated by leucine supplementation, but MHC may be less sensitive to anabolic factors with aging. PMID- 15130670 TI - Long-term treatment with antioxidants and a program of behavioral enrichment reduces age-dependent impairment in discrimination and reversal learning in beagle dogs. AB - The effects of long-term treatment with both antioxidants and a program of behavioral enrichment were studied as part of a longitudinal investigation of cognitive aging in beagle dogs. Baseline performance on a battery of cognitive tests was used to assign 48 aged dogs (9-12 years) into four cognitively equivalent groups, of 12 animals per group: Group CC (control food-control environment), group CE (control food-enriched environment); Group AC (antioxidant fortified food-control environment); Group AE (fortified food-enriched environment). We also tested a group of young dogs fed the control food and a second group fed the fortified food. Both groups of young dogs received a program of behavioral enrichment. To evaluate the effects of the interventions on cognition after 1 year, the dogs were tested on a size discrimination learning task and subsequently on a size discrimination reversal learning task. Both tasks showed age-sensitivity, with old dogs performing more poorly than young dogs. Both tasks were also improved by both the fortified food and the behavioral enrichment. However, in both instances the treatment effects largely reflected improved performance in the combined treatment group. These results suggest that the effectiveness of antioxidants in attenuating age-dependent cognitive decline is dependent on behavioral and environmental experience. PMID- 15130671 TI - Hormonal control of the yolk precursor vitellogenin regulates immune function and longevity in honeybees. AB - A striking example of plasticity in life span is seen in social insects such as ants and bees, where different castes may display distinct ageing patterns. In particular, the honeybee offers an intriguing illustration of environmental control on ageing rate. Honeybee workers display a temporal division of labour where young bees (or 'hive bees') perform tasks within the brood nest, and older bees forage for nectar, pollen propolis and water. When bees switch from the hive bee to the forager stage, their cellular defence machinery is down-regulated by a dramatic reduction in the number of functioning haemocytes (immunocytes). This study documents that the yolk precursor vitellogenin is likely to be involved in a regulatory pathway that controls the observed decline in somatic maintenance function of honeybee foragers. An association between the glyco-lipoprotein vitellogenin and immune function has not previously been reported for any organism. Honeybee workers are functionally sterile, and via the expression of juvenile hormone, a key gonotrophic hormone in adult insects, their vitellogenin levels are influenced by social interactions with other bees. Our results therefore suggest that in terms of maintenance of the cellular immune system, senescence of the honeybee worker is under social control. PMID- 15130672 TI - The variations during the circadian cycle of liver CD1d-unrestricted NK1.1+TCR gamma/delta+ cells lead to successful ageing. Role of metallothionein/IL 6/gp130/PARP-1 interplay in very old mice. AB - NKT cells derive from the thymus and home to the liver. Liver NKT cells can be divided in two groups: 'classical' and 'non-classical'. The first is CD1d restricted, the second is CD1d-unrestricted. NKT cells (classical and non classical) co-express T-cell receptor (TCR) and NK-cell marker (NK1.1), display cytotoxicity and produce IFN-gamma under IL-12 stimulation affecting, thereby, Th1 response and innate immunity. NK1.1(+)TCR alpha/beta(+) cells belong to both groups. NK1.1(+)TCR gamma/delta(+) cells belong to the second group. Anyway, both NKT cell subtypes, via IFN-gamma production, protect against viruses and bacteria from early in life. Immune variations as well as zinc rhythmicity during the circadian cycle confer the immune plasticity, which is essential for successful ageing. Liver NK1.1(+)TCR gamma/delta(+) cells, rather than TCR alpha/beta(+), from young and very old mice display 'in vitro' (under IL-12 stimulation) nocturnal peaks in cytotoxicity and IFN-gamma production. The acrophase of liver NK1.1(+)TCR gamma/delta(+) cells is present in young and very old mice, not in old. The interplay among zinc-bound metallothionein (MT)/IL-6/gp130/poly(ADP ribose) polymerase-1 (PARP-1) may be involved in conferring plasticity to liver NK1.1(+)TCR gamma/delta(+) cells. IL-6, via sub-unit receptor gp130, induces MTmRNA. At night, gene expressions of MT, IL-6, gp130 are lower in very old mice than old and young MT-I transgenic mice (MT-I*). In very old mice, this phenomenon allows limited sequester of intracellular zinc from MT leading to good free zinc ion bioavailability for immune efficiency and zinc-dependent PARP-1 activity. Indeed (1) in vitro, high IL-6 provokes strong accumulation of MT, impaired cytotoxicity and low zinc ion bioavailability in liver NK1.1(+)TCR gamma/delta(+) cells exclusively from old and MT-I* mice. (2) The ratio total/endogen PARP-1 activity is higher in very old than in old and MT-I* mice, suggesting a higher capacity of PARP-1 in base excision DNA-repair in very old age thanks to low zinc-bound MT. Cytotoxicity and IFN-gamma production from liver NK1.1(+)TCR gamma/delta(+) cells are thus preserved leading to successful ageing. In conclusion, MT/IL-6/gp130/PARP-1 interplay may confer plasticity to liver CD1d unrestricted NK1.1(+)TCR gamma/delta(+) cells, where MT, IL-6, gp130 are the main upstream protagonists, and PARP-1 is the main downstream protagonist in immunosenescence. PMID- 15130674 TI - Hypothetical soluble KIR2DS4 natural killer cell receptor molecule does not associate with successful ageing in the Irish. AB - The identification of immunogenetic longevity markers is a major area of molecular gerontological research. A number of genetic loci have been examined, e.g. the HLA and cytokine networks. This study investigated a genetic marker within the highly polymorphic KIR gene system with successful ageing in the Irish population. A 22 bp deletion was identified in the KIR2DS4 gene that predicts a truncated soluble KIR molecule with one intact Ig-like domain. The frequency of this variant was determined using a specific-primer PCR methodology. There was no observed association between this common polymorphic variation within this activatory KIR gene and the aged Irish population. This is the first study of KIR polymorphism in ageing and although no association was identified, the importance of the KIR network in the immune response and its polymorphic nature warrants more detailed analysis to ascertain its role in immunosenescence. PMID- 15130673 TI - Autolysosomes accumulate during in vitro CD8+ T-lymphocyte aging and may participate in induced death sensitization of senescent cells. AB - As autophagic inclusions accumulate in senescent fibroblasts, we wondered whether an increase in cellular fragility during in vitro lymphocyte aging may be related to an autolysosome accumulation. We established that, during long-term cultures, repeatedly stimulated T-lymphocytes acquired characteristics of replicative senescence and became progressively intolerant to activation. Cell death following stimulations: (i) corresponded to apoptosis, associated with necrosis at the end of the culture; (ii) was not, for its main part, mediated through CD95/CD178 or TNFRII/TNF alpha interactions; and (iii) occurred in spite of bcl-2 increased expression. After 14 weeks of culture, the percentage of lymphocytes containing at least one autophagic inclusion (p<0.0001) and the lipofuscin autofluorescence in lymphocytes (p<0.0001) were significantly increased. The expression of several genes regulating autophagy did not significantly vary with the age of the culture. Forty-eight hours after each stimulation, the percentage of induced cell death rose while, in the remaining living cells, the percentage of lymphocytes with autophagic vacuoles (p<0.05), with beta-galactosidase activity and the lipofuscin autofluorescence (p<0.001) significantly decreased, suggesting the preferential death of cells with autophagy. Our data support the view that the accumulation of autolysosomes in senescent lymphocytes might aggravate cellular fragility, leading to apoptosis and necrosis mainly induced by lymphocyte activation. PMID- 15130675 TI - Is early life body weight a predictor of longevity and tumor risk in rats? AB - Heavy body weight (BW) is thought to be associated with reduced longevity and age associated diseases, including cancer, both in laboratory rodents and humans. To further investigate the interactions between BW, longevity and spontaneous tumor development, we measured the correlations between BW in early life, BW in middle life, and parameters of life span and tumorigenesis in male and female outbred rats. The data show that BW at the ages of both 3 and 12 months are significant predictors of longevity in rats. Heavier female rats tend to live longer than the lighter female rats, while in male those who were light at 3 months but heavy at 12 month had the best longevity. BW at the age 3 months was not predictive of tumor growth but being heavier at the age of 1 year did confer an increased risk of tumor development for both male and female rats. PMID- 15130676 TI - Effects of paradoxical sleep deprivation on blood parameters associated with cardiovascular risk in aged rats. AB - The effects of 96 h of paradoxical sleep deprivation (PSD) on blood parameters associated with cardiovascular risk were studied in young (3-month old) and aged (22-month old) rats. In general, aging was associated with an overall increase in most measures, irrespective of sleep deprivation condition. The latter manipulation also had significant effects on blood variables, but not in a consistent pattern. Thus, PSD significantly reduced triglyceride levels in both young and aged rats; it reduced blood viscosity in aged but not in young rats, and had no effect on the increased cholesterol levels observed in aged controls. Examinations of cholesterol fractions revealed significant increases in low density lipoprotein and high density lipoprotein in aged PSD rats compared to respective controls, whereas very low density lipoprotein was significant decreased after PSD in both young and aged animals. PSD increased vitamin B(12) levels in aged rats, and significantly decreased homocysteine levels in young but not in aged rats which in turn were already reduced. Folate levels were the only variable that was unaffected by aging and/or PSD. These results indicate that PSD has significant but heterogeneous physiological effects in aged rats and may intensify certain aging-related effects which contribute to cardiovascular disease risk while attenuating others. PMID- 15130677 TI - Influence of age and vitamin E on post-ischemic acute renal failure. AB - The aging process causes progressive deterioration in kidney structure and function. Aberrant generation of reactive oxygen species has been implicated in both age-related and ischemia-related tissue injury. Vitamin E (VE), one of the most powerful and effective exogenous antioxidants, prevents lipid peroxidation and protects against the effects of oxidative stress. The objective of this study was to determine the influence of age and VE on post-ischemic acute renal failure (ARF). Young adult, middle-aged and aged male Wistar rats were maintained on three different 30-day diets: Normal, VE absent and VE supplemented. On day 30, urinary protein and serum cholesterol and VE were measured. On day 31, rats were subjected to 60' clamping of the left renal artery plus right nephrectomy. Inulin clearance (InCl) was performed 48 h after renal ischemia. Malondialdehyde (MDA) was measured in the cortex of normal and 48-h post-ischemic kidneys. Urinary protein and serum cholesterol were higher in aged rats than in other rats. With aging, InCl decreased progressively. Vitamin E deficiency aggravated ARF. In middle-aged and aged rats, VE supplementation protected against ARF. In the absence of VE, MDA increased with age. In conclusion, our data suggest that ARF becomes more severe with age and that ischemia/reperfusion injury is exacerbated when antioxidant-scavenging ability of the kidney is impaired by VE deficiency. Supplementation with VE is essential for protecting aging kidneys against ischemic ARF. PMID- 15130679 TI - Are age-related deficits in balance performance mediated by time of day in a prosimian primate (Microcebus murinus)? AB - We examined the effects of age and time of day on balance performance in the gray mouse lemur (Microcebus murinus) in relation to body temperature variations. Groups of young, adult and aged animals were entrained to either short (SP, L/D 10/14) or long photoperiod (LP, L/D 14/10) and tested in the accelerating Rotarod at three phases of their light/dark cycle (beginning and end of the light phase, beginning of the dark phase). In addition, for each test, rectal temperature (Tr) was measured. Under LP, whatever the test phase, this primate showed a clear age related decrease in balance performance. Under SP, no significant age-related decline in balance could be detected. Whatever the photoperiod, an effect of time of day on balance performance could only be seen in adults, with better performances at the beginning of the dark phase when Tr values were higher. Under LP, daily variations of both balance performance and Tr disappeared with advancing age. Consequently, age differences were substantially greater when testing was conducted during the dark phase of the light/dark cycle. The time of day effect on balance performance and the loss of daily variations with age suggest the influence of age-related changes within the biological clock. PMID- 15130678 TI - Function and clustered expression of MaxiK channels in cerebral myocytes remain intact with aging. AB - The incidence of stroke increases significantly in the aging population where stroke related deaths boost at >75 years and survivors are often permanently disabled. Aging is known to decrease cerebral blood flow likely due to an increase in arterial tone. Although MaxiK channels are key regulators of cerebral arterial tone their pattern of expression and function in cerebral blood vessels during aging is unknown. Using specific antibodies against the alpha-subunit of MaxiK channels and current recordings, we now demonstrate that in aging cerebral myocytes, MaxiK channels remain healthy. Furthermore, we show for the first time that in the vasculature, MaxiK channels are expressed in clusters. Clusters have an estimated radius of approximately 200 nm in young rats (3-5 month old Fisher 344 rats) which remains normal in old (25-30 month rats) cerebral myocytes. Consistent with a healthy MaxiK channel expression in old cerebral arteries, MaxiK current density, kinetics and Ca(2+) sensitivity were practically identical in young and old myocytes. Sensitivity to nanomolar concentrations of dehydrosoyasaponin-I that activates channels formed by alpha and beta subunits is also the same in young and old myocytes. These results demonstrate that MaxiK channels maintain normal expression during cerebral aging which is in sharp contrast to our previous finding of loss of expression in aging coronary arteries. It seems therefore, that cerebral myocytes have developed a protective anti-aging mechanism leading to the continued expression of MaxiK channels. PMID- 15130680 TI - Analysis of premature centromere division (PCD) of the X chromosome in Alzheimer patients through the cell cycle. AB - Cytogenetic analysis of the X chromosome in phytohaemagglutinin stimulated peripheral blood lymphocytes was evaluated in 12 sporadic Alzheimer disease (AD) patients and in 11 healthy subjects. For chromosome analysis two methods were used: (1) standard analysis of G-banded metaphase chromosomes and; (2) fluorescent in situ hybridization (FISH) for the detection of the X chromosome centromeric region in interphase nuclei. Cytogenetic analysis revealed that the X chromosome expresses premature centromere division (PCD) in AD females in 10.53% of metaphase cells and in 15.22% of interphase nuclei. In AD men the percentages were 3.98 and 6.06%, respectively. X chromosome PCD in the female control group showed a percentage of 7.46% in metaphase cells and 9.35% in interphase nuclei and in male controls the percentages were 2.84% in metaphases and 5.54% in interphase nuclei. The results of FISH analysis showed that PCD could occur much earlier than metaphase of mitosis, i.e. in interphase of the cell cycle, immediately after replication. The FISH method can be used for PCD verification in all phases of the cell cycle in various disorders including AD. PMID- 15130681 TI - Rapid telomere attrition in cardiac tissue of the ageing Wistar rat. AB - Many studies show an association between ageing and mean telomere length in DNA isolated from peripheral blood mononuclear cells, few studies have examined less accessible tissues. This study has two objectives: (i) to define the best method to prepare rodent DNA for telomere length measurement by Southern blotting and (ii) to determine whether there are differential rates of telomere attrition in different rodent tissues. We found that the use of agarose plugs for DNA isolation was essential for the accurate measurement of rodent telomere length. Tissue was collected from neonatal (3 days) or aged (18-24 months) male Wistar rats and terminal restriction fragment (TRF) length was measured by Southern blotting. Cardiac tissue from aged rats showed a 38% loss of TRF length compared with newborn animals (p<0.001, n=13), this contrasts with much smaller reductions in brain (1.6%), liver (14.2%), kidney (8.9%) and lung (9.7%). This study demonstrates that the methods of DNA preparation are critical for accurate measurement of telomeres in rodent tissues. Moreover, we show differential rates of telomere attrition in rat tissues, the heart being most susceptible to telomere loss. These observations could have important implications for the study of age-specific changes in tissue function. PMID- 15130682 TI - Genome size is not correlated positively with longevity in fishes (or homeotherms). PMID- 15130684 TI - IFN-gamma production by CMV-specific CD8+ T cells is high in elderly donors. PMID- 15130686 TI - Genes and mutations in human idiopathic epilepsy. AB - Thirteen genes have already been identified in human idiopathic epilepsies since 1995, but they account only for a minority of all epilepsy cases. Most of these genes are associated with rare monogenic epilepsy syndromes, but some of them contribute to the common epilepsy subtypes. The questions remains to be answered how many more epilepsy genes exist in brain. Idiopathic epilepsies are common neurological disorders, and it can therefore be expected that the total number of genes associated with an increased seizure susceptibility is much higher than 13. Most of the known genes code for either voltage-gated or ligand gated ion channels, but recently two epilepsy genes have been found which do not fit into the concept of epilepsies as channelopathies. It can therefore be suspected that more than one pathogenetic concept exists in epileptogenesis. PMID- 15130687 TI - Influence of orthopaedic-technical aid on the kinematics and kinetics of the knee joint of patients with neuro-orthopaedic diseases. AB - In our gait laboratory, the gait pattern of 18 youths with neurogenic foot deformities as a result of spina bifida or cerebral palsy was examined. The influence of technical orthopaedic devices for the foot and ankle on kinematics and kinetics of the gait and especially of the knee joint were analyzed. Kinematic data were derived from 3D-video analysis, kinetic data from force plates and pressure distribution plates. Muscle activities were measured with eight-channel EMG. The data were examined to see if there were differences when using the technical devices. All patients had already been supported externally with the different devices like orthopaedic shoes, insoles, Nancy Hilton orthoses and orthoses for the lower leg extremity. The devices restricted to the foot and the ankle joint improved the feeling of gait stability of the patients. In this investigation, the different supports had various but little effects on the kinematics and kinetics of the knee joint, depending on the type of orthoses used and the kind of handicap of the youths. Because of the great expenditure, the data obtained in this study were taken from a small number of patients. Therefore, they are individual findings and are of restricted general significance. PMID- 15130688 TI - Clinical profiles of children with cerebral palsy having lesions of the thalamus, putamen and/or peri-Rolandic area. AB - Neurological findings, motor symptoms, mental abnormality and dysarthria were examined in 28 children with lesions in the thalamus, putamen, and/or peri Rolandic area. The thalamus and putamen were involved in eight, and only the thalamus in ten of the children. Most of these 18 children had mild disabilities; they did not have severe mental retardation and could walk alone, speak words, and grasp an object. Dominant flexion of the hips was observed in many of the children who could walk. Two-thirds of these children had athetotic involuntary movement and the remaining had gross or fine motor abnormalities although they had no involuntary movement. In most of these children, reaching patterns were abnormal and were affected by shoulder retraction. Their abnormal movements were thought to be inappropriate muscle activity brought about by voluntary movements. In the remaining ten children, the thalamus, putamen, and peri-Rolandic area were all involved. Many had severe disabilities such as severe mental retardation and the inability to sit, speak words, or grasp an object. All had athetotic involuntary movements. Three children had spasticity of the lower extremities. Five children with severe disabilities and no spasticity were thought to have apparent weakness with athetosis. PMID- 15130689 TI - Sleep disturbances in Angelman syndrome: a questionnaire study. AB - Only few studies are available on sleep disorders in Angelman syndrome (AS), a neurodevelopmental disorder with several behavior disturbances. The aim of this study was to determine the prevalence of sleep disorders in a relatively large group of AS subjects, compared to that of age-matched controls. Forty-nine consecutive parents of patients with AS (26 males and 23 females aged 2.3-26.2 years) were interviewed and filled out a comprehensive sleep questionnaire. Based on their genetic etiology, four groups were defined: deletion of chromosome 15q11 13 (25 subjects); methylation imprinting mutation (six subjects), UBE3A mutations (seven subjects) and paternal uniparental disomy (five subjects). In the remaining cases genetic testings were negative. A significantly high frequency of disorders of initiating and maintaining sleep, prolonged sleep latency, prolonged wakefulness after sleep onset, high number of night awakenings and reduced total sleep time were found in our AS patients, as compared to age-matched controls. We also found other types of sleep disorders, never reported before, such as enuresis, bruxism, sleep terrors, somnambulism, nocturnal hyperkinesia, and snoring. No differences were found between the four genetic aetiology groups. Moreover, we did not find important improvement of sleep disturbances from pre pubertal to post-pubertal ages. Our data confirm the significant presence of sleep/wake rhythms fragmentation, peculiar of AS, and also demonstrate the presence of several other types of sleep disturbances in this syndrome. PMID- 15130690 TI - Treatment and outcome in patients with febrile convulsion associated with epileptiform discharges on electroencephalography. AB - The aim of this study is to determine the efficacy of prophylactic treatment for patients with febrile convulsions (FCs) in whom electroencephalograms (EEGs) revealed epileptiform discharges. We retrospectively investigated 43 patients who met the following criteria: (a) at least one FC during the study period; (b) epileptiform discharges were first recognized; (c) no unevoked seizures before epileptiform discharges were first seen; (d) normal psychomotor development and no neurological abnormality; and (e) follow-up >3 years. The clinical characteristics, treatment, and a later occurrence of FCs or unevoked seizures were studied. EEGs revealed focal epileptiform discharges in 25 patients and generalized ones in 18. There was no significant difference in the rate of recurrence of FC or occurrence of unevoked seizures between those with focal and generalized epileptiform discharges. No prophylaxis was performed in ten patients, 14 patients being treated with intermittent diazepam and 19 with a daily anticonvulsant. The rate of recurrence of FC was not significantly different between patients with and without prophylaxis. Unevoked seizures were only observed in two patients undergoing daily treatment. Intermittent or daily anticonvulsant therapy will not reduce the risk of recurrence of FCs or later development of unevoked seizures in patients with FC with epileptiform discharges. PMID- 15130691 TI - Increased NPC1 mRNA in skin fibroblasts from Niemann-Pick disease type C patients. AB - Niemann-Pick disease type C (NP-C) is an autosomal recessive lipid-storage disease that is characterized by progressive neurodegeneration and hepatosplenomegaly. Since identification of the NPC1 gene in 1997, a total of 120 disease-causing mutations have been reported. In this study, two novel mutations were identified, namely c.2508[-2509]A del (837Fs-838X) in exon 16 and T3194G (V1065G) in exon 21. To explore the impact of NPC1 mutations on transcription of this gene, we analyzed NPC1 mRNA levels in skin fibroblasts derived from NP-C patients. Fibroblasts from patients with missense mutations showed increased levels of NPC1 mRNA while fibroblasts from patients with a specific frameshift mutation showed mRNA levels similar to those of normal control subjects. These results suggest that NPC1 transcription levels are altered in cells with mutations in the NPC1 gene. PMID- 15130692 TI - Visual perceptual impairment in children at 5 years of age with perinatal haemorrhagic or ischaemic brain damage in relation to cerebral magnetic resonance imaging. AB - Children with perinatally acquired parenchymal haemorrhage are thought to have better visual perceptual skills than those with leukomalacia. We examined seven prematurely born children with parenchymal haemorrhage and 14 with grades 2-4 leukomalacia, at the age of 5 years. Clinical and magnetic resonance imaging parameters were related to visual perceptual performance assessed with the L94, using performance age. Belonging to the leukomalacia group, the inability to walk, a diminished peritrigonal white matter, a high degree of gliosis and cortical damage were associated with poorer visuo-perceptual skills. Enlarged lateral ventricles, confirming the findings of Melhelm (Radiology 214 (2000) 199), were associated with both cognitive, perceptual and motor problems and probably reflect the considerable extent of the brain damage. Specific factors protecting against visual perceptual impairment were a preserved volume of the right optical radiation and of the splenium of the corpus callosum. Children with leukomalacia are at considerable risk of visual perceptual impairment. Children with right-sided parenchymal haemorrhages also appear to be at risk although they function much better due to better motor and cognitive skills. PMID- 15130693 TI - Clinical and electroencephalographic characteristics of children with febrile seizures plus. AB - OBJECTIVES: Febrile seizures plus (FS+) are attracting attention for their corresponding genetic abnormalities, and are defined as febrile seizures (FS) continuing beyond 6 years of age (late FS) or those associated with afebrile seizures. We tried to elucidate their clinical and EEG characteristics as compared with those of children having only FS. SUBJECTS AND METHODS: We reviewed clinical records in a pediatric neurology clinic to identify 31 patients with FS+ (group FS+) and 51 with only FS (group FS). Their family history of seizures, clinical features and EEG findings were compared. RESULTS: A family history of seizures was noted in 14 patients (45.2%) of group FS+ and in 24 (47.1%) of group FS. In group FS+, 19 patients had late FS, 11 had afebrile seizures, and the remaining one had both types of seizures. Two patients had seizures induced by TV/video-game as well, and another suffered from absences. Epileptic EEG abnormalities, which included diffuse spike-waves and focal spikes, were noted in 13 patients (41.9%) of group FS+ and 12 (23.5%) of group FS. CONCLUSIONS: The clinical and EEG characteristics of the children having FS+ were diverse, without significant differences from those with FS except for the seizures types. PMID- 15130694 TI - A rare complication of generalized edema in juvenile dermatomyositis: a report of one case. AB - Juvenile dermatomyositis (JDM) is a rare autoimmune disease characterized by inflammation of the muscle, connective tissue, skin, gastrointestinal tract and small nerves. Periorbital and facial edema may also be associated. Although localized edema is a common feature of juvenile dermatomyositis, generalized edema has been reported rarely. In this article, we report a 14-year-old boy with juvenile dermatomyositis presenting with generalized edema. Of the diagnostic criteria of JDM, severe symmetric weakness of the proximal musculature, characteristic cutaneous changes, elevated serum muscle enzymes and myopathic electromyographic abnormalities were observed. Magnetic resonance imaging (MRI) of the lower extremities and pelvis showed marked diffuse edema in the subcutaneous tissue, muscles and myofascia. We suggest that MRI findings, which are not among the diagnostic criteria, may also be included in the diagnostic criteria of JDM. To the best of our knowledge, this is the 19th case of JDM reported for generalized edema in the English literature. PMID- 15130695 TI - Unusual neurophysiological features in Cockayne's syndrome: a report of two cases as a contribution to diagnosis and classification. AB - To clarify the diagnostic value of neurophysiological investigations in patients with Cockayne's syndrome (CS). The study involved two patients with clinical diagnoses of classical and severe CS, who were neurophysiologically evaluated by means of: (1) multimodal visual (VEPs), brainstem auditory (BAEPs) and upper limb somatosensory (SEPs) evoked potentials; (2) electroretinography; and (3) nerve conduction and needle electromyography studies. Both patients showed multimodal evoked potential (EP) signs of central nervous system involvement that overlapped in severity and extent, and were consistent with demyelination along the central sensory pathways. Flash VEPs and SEPs were more altered than pattern VEPs and BAEPs. No signs of retinopathy or hearing loss of cochlear origin were detected. The nerve conduction and needle electromyography studies showed severe signs of sensory and motor demyelinating and axonal peripheral neuropathy. Peripheral neuropathy was clinically uncertain. There were no significant differences between the two patients. Our results show that combined multimodal EP and nerve conduction studies are diagnostically highly sensitive even in the early stage of CS, but their ability to distinguish classical and severe CS is limited. The unusual features were characterised by the absence of clinical and electrophysiological signs of otherwise common retinopathy and neurosensory hearing loss. BAEPs seem to be more useful than VEPs or SEPs in the diagnostic work-up of patients with suspected CS. PMID- 15130696 TI - NK-3 receptor activation depolarizes and induces an after-depolarization in pyramidal neurons in gerbil cingulate cortex. AB - The involvement of tachykinins in cortical function is poorly understood. To study the actions of neurokinin-3 (NK3) receptor activation in frontal cortex, whole cell patch clamp recordings were performed from pyramidal neurons in slices of cingulate cortex from juvenile gerbils. Senktide (500nM), a selective NK3 receptor agonist, induced a transient increase in spontaneous EPSPs in layer V pyramidal neurons, accompanied by a small depolarization ( approximately 4 mV). EPSPs during senktide had a larger amplitude and faster 10-90% rise time than during control. Senktide induced a transient depolarization in layer II/III pyramidal neurons, which often reached threshold for spikes. The depolarization ( approximately 6 mV) persisted in TTX, and was accompanied by an increase in input resistance. Senktide also transiently induced a slow after-depolarization, which appeared following a depolarizing pulse. The slow after-depolarization persisted in TTX. These data suggest that activation of NK3 receptors on layer II/III pyramidal neurons induce post-synaptic depolarization and an after depolarization, which could be mediated by blockade of a leak potassium conductance and a non-selective cation conductance, respectively. PMID- 15130697 TI - GABAC receptor mediated inhibition in acutely isolated neurons of the rat dorsal lateral geniculate nucleus. AB - In the dorsal lateral geniculate nucleus (dLGN), GABA(C) receptors seems to be specifically expressed by local GABAergic interneurons. Although the presence of GABA(C) receptors has been demonstrated, a quantitative estimation of their contribution to inhibition in dLGN is lacking. Because the amount of inhibition mediated by these receptors might reflect their functional importance we performed whole-cell patch clamp recordings from dLGN cells acutely dissociated from brain slices. We focally applied the GABA receptor agonist muscimol and quantified effects mediated through either GABA(C) or GABA(A) receptors. Because their basic dendritic morphology was preserved, we tried to morphologically differentiate between thalamocortical cells and local interneurons. In the majority of multipolar cells, representing thalamocortical projection neurons, the specific GABA(A) receptor antagonist bicuculline completely blocked muscimol induced currents. In contrast, in most of the bipolar cells, representing interneurons, bicuculline blocked only 70-80% of the muscimol induced currents. The remaining currents were blocked by co-application of TPMPA, a specific GABA(C) receptor antagonist, or picrotoxin, an unspecific GABA(A) and GABA(C) receptor blocker. The latter neurons were also sensitive to the selective GABA(C) receptor agonist cis-aminocrotonic acid. These results indicate that in those dLGN neurons that express GABA(C) receptors, these receptors contribute considerably to GABAergic inhibitory inputs. PMID- 15130698 TI - Attenuation of mechanical but not thermal hyperalgesia by electroacupuncture with the involvement of opioids in rat model of chronic inflammatory pain. AB - Opioid peptides have been proven effective in reducing the sign of hyperalgesia associated with inflammation. Electroacupuncture (EA) produces antinociception via release of endogenous opioid peptides in normal rats. Moreover, intrathecal injection of dynorphin has antinociceptive effect in rats. The present study was designed to examine whether EA has effect on the thermal and mechanical hyperalgesia in rat model of complete Freund's adjuvant (CFA)-induced inflammatory pain. The results are the following: (1) single session of 100Hz EA (0.5-1.0-1.5 mA, 10 min for each intensity) at both Zusanli (ST 36) and Sanyinjiao acupoints (SP 6) significantly increased mechanical withdrawal threshold determined by von Frey filaments but not with thermal withdrawal latency that is determined by hot plate (52 +/- 0.2 degrees C); (2) 100 Hz EA applied twice a week for 4 weeks and showed a significant decrease in the mechanical hyperalgesia at the third and fourth week, with no effect on thermal hyperalgesia; (3) naloxone (20 mg kg(-1)) had the ability to reverse the inhibition of the mechanical hyperalgesia produced by a single session of EA. In conclusion, the present results indicate that a single or repetitive EA could reduce mechanical hyperalgesia, but not thermal hyperalgesia, in CFA-inflammatory pain rats, and the opioid system might be involved in these effects. PMID- 15130699 TI - Intracerebral cytokine profiles in adult rats grafted with neural tissue of different immunological disparity. AB - To understand graft rejection in cell based therapies for brain repair we have quantified IL-1beta, IL-2, IL-4, IL-10, IL-12p40, IFN-gamma and TNF-alpha mRNA levels using real-time PCR, at days 4, 14, and 42 post-transplantation, in rats engrafted with syngeneic, allogeneic, concordant and discordant xenogeneic neural tissues. In addition, in the discordant xenografts immunohistochemistry and in situ hybridization were applied to detect local expression of IFN-gamma, TNF alpha, IL-10 and TGF-beta. Allografts remained non-rejected but expressed IL 1beta, TNF-alpha and IL-4 transcripts but not IL-12p40 and IFN-gamma. Xenografts demonstrated distinct cytokine profiles that differed from syngeneic and allogeneic grafts. Non-rejected discordant xenografts contained higher levels of TNF-alpha transcripts and lower levels of IL-2 transcripts than the rejected ones at day 42. Discordant xenografts displayed a stronger and earlier expression of IL-1beta and TNF-alpha, followed by T-helper 1 and T-helper 2 associated cytokine expression. The number of cells expressing mRNA encoding TNF-alpha and TGF-beta was significantly increased over time in the discordant group. In conclusion, the immunological disparity of the implanted tissue explains survival rates and is associated with different cytokine profiles. In allografts, a chronic inflammatory reaction was detected and in xenogeneic grafts a delayed hypersensitivity like reaction may be involved in rejection. PMID- 15130700 TI - Primary structure of guinea pig preprodynorphin and preproenkephalin mRNAs: multiple transcription initiation sites for preprodynorphin. AB - Preprodynorphin and preproenkephalin are protein precursors from which are derived two classes of opioid neurotransmitter peptides. Dynorphin A((1-17)) is produced by proteolytic processing of prodynorphin, and processing of proenkephalin yields the enkephalin peptides. We report here on the isolation and sequencing of multiple clones for these two mRNAs from a cDNA library. Two cDNA clones of preprodynorphin contained the full-length sequence (2.35 kb) with the primary structure predicted from the guinea pig gene sequence. In contrast, one clone encoded the full-length sequence but also an additional 192 nt at the 5' end. This sequence has high homology to the 5' flanking region of the human preprodynorphin gene, and RNase protection assays demonstrated that in addition to a primary initiation site, transcription of this mRNA is initiated at several sites 160-190 nt 5' with respect to the primary site. This difference may alter translational efficiency or mRNA stability. The sequence of preproenkephalin cDNA clones confirmed the structure predicted from the gene sequence. One clone, however, contained sequences encoded by exons 2 and 3, and initiated within the first intron (intron A) of the gene. We used RNase protection mapping to assess the abundance in the brain and pituitary of preproenkephalin transcripts that initiate within intron A. These studies confirmed that the primary transcription start site is 28 nucleotides downstream from the TATAA site, and that intron A sequences are not present in significant amounts in these tissues. PMID- 15130701 TI - NMDA dilates pial arteries by KATP and Kca channel activation. AB - This study was designed to characterize the role of ATP sensitive (K(ATP)) and calcium sensitive (K(ca)) K(+) channel activation in N-methyl-d-aspartate (NMDA) induced pial artery dilation in newborn pigs equipped with a closed cranial window. NMDA (10(-8), 10(-6)M) elicited pial artery dilation that was blunted by the K(ATP) channel antagonist glibenclamide (10(-6)M) and attenuated by the K(ca) channel antagonist iberiotoxin (10(-7)M) (8 +/- 1% and 14 +/- 1% versus 2 +/- 1% and 4 +/- 1% versus 5 +/- 1% and 9 +/- 2% for NMDA in the absence and presence of glibenclamide or iberiotoxin, respectively). Combined administration of glibenclamide and iberiotoxin essentially eliminated the response. Similar results were observed for glutamate. These data show that NMDA elicits cerebrovasodilation through activation of K(ATP) and K(ca) channels in newborn pigs. PMID- 15130702 TI - Metallothionein isoforms (I+II and III) and interleukin-6 in the hippocampus of old rats: may their concomitant increments lead to neurodegeneration? AB - Metallothionein (MT)-III isoform is a brain metal-binding protein that, like the MT-I + II isoform, binds zinc with high affinity. In the young-adult age, MT-III isoform increases during transient stress while MT-I + II isoform decreases, suggesting compensatory phenomena between the two isoforms and a protective role of MT-III against oxidative damage. This role may be questioned during ageing, because the stress-like condition is chronic in ageing due to high persistent levels of interleukin-6. In the present study, high expression of MT-III and MT-I + II genes (examined by RT-PCR and in situ hybridisation) was found in the hippocampus of old rats. These results indicate that a large amount of free zinc ions can be sequestered by MT isoforms, leading to impaired zinc-dependent functions in the ageing brain. In addition, zinc (tested with the Timm's method) was found to be low in mossy fibres from the old hippocampus. As this method tests bound and unbound zinc, we also investigated free zinc ion bioavailability based on the ratio active thymulin/total thymulin. We found that zinc ion bioavailability was low in old rats, together with increased interleukin-6 mRNA, high expression of both MT isoforms and reduced number of synapses whose function is zinc-dependent, in the old hippocampus. The results indicate that concomitant increments of both MT isoforms may provoke detrimental synergistic effects leading to reduced free zinc ion bioavailability for synapses. As a consequence, compensatory phenomena between MT isoforms may not occur in the old hippocampus due to chronic stress-like condition elicited by high persistent levels of interleukin-6. PMID- 15130703 TI - Inhibitory effect of iodophenpropit, a selective histamine H3 antagonist, on amygdaloid kindled seizures. AB - The effect of histamine H(3) antagonist, iodophenpropit on amygdaloid kindled seizures in rats was studied in comparison with those of other H(3) antagonists. Under pentobarbital anesthesia, the rats were fixed to a stereotaxic apparatus and bipolar electrodes were implanted into the amygdala. Electrodes were connected to a miniature receptacle, which was embedded in the skull with dental cement. To cause kindled seizures, electrical stimulation was applied to the amygdala bipolarly every day by a constant current stimulator, and electroencephalogram and convulsive behavior were observed. Drug effects were estimated in rats showing generalized kindled seizures. Intraperitoneal injection of H(3) antagonists, iodophenpropit, thioperamide, AQ0145 and clobenpropit, resulted in a dose-related inhibition of amygdaloid kindled seizures. The effect of iodophenpropit on amygdaloid kindled seizures was more potent than those of thioperamide, AQ0145 and clobenpropit. In conclusion, iodophenpropit may be useful for the treatment of partial epilepsy and/or secondary generalized seizures in humans. PMID- 15130704 TI - Resting state brain glucose metabolism is not reduced in normotensive healthy men during aging, after correction for brain atrophy. AB - Studies using positron emission tomography (PET) have reported that global and regional values for cerebral blood flow and metabolic rates for glucose (CMRglc and rCMRglc) decline with age in humans. We wished to determine if such decreases could have reflected a partial volume effect (PVE) of cerebral atrophy in the elderly, rather than "intrinsic" reductions per gram brain. We used PET to compare rCMRglc, before and after correcting for the PVE, between 13 healthy older men (aged: 55-82 years) and 11 healthy young men (aged: 22-34 years). PET was performed with 18F-fluoro-2-deoxy-d-glucose while the subjects were in the "resting" state (eyes covered and ears plugged with cotton). The PET scans were normalized to a common brain volume after superimposing them on the subjects' tissue segmented magnetic resonance scans. Analysis showed that rCMRglc in the absence of a PVE correction was significantly less in the older group in insular, frontal, superior temporal cortical, and thalamic regions. Statistical significant differences in rCMRglc, however, were absent after the PVE correction. Thus, statistically significant age reductions in regional brain glucose metabolism, corrected for brain atrophy, are not detectable in healthy normotensive men scanned while in the resting state. PMID- 15130705 TI - Neuronal density, size and shape in the human anterior cingulate cortex: a comparison of Nissl and NeuN staining. AB - There are an increasing number of quantitative morphometric studies of the human cerebral cortex, especially as part of comparative investigations of major psychiatric disorders. In this context, the present study had two aims. First, to provide quantitative data regarding key neuronal morphometric parameters in the anterior cingulate cortex. Second, to compare the results of conventional Nissl staining with those observed after immunostaining with NeuN, an antibody becoming widely used as a selective neuronal marker. We stained adjacent sections of area 24b from 16 adult brains with cresyl violet or NeuN. We measured the density of pyramidal and non-pyramidal neurons, and the size and shape of pyramidal neurons, in laminae II, III, Va, Vb and VI, using two-dimensional counting methods. Strong correlations between the two modes of staining were seen for all variables. However, NeuN gave slightly higher estimates of neuronal density and size, and a more circular perikaryal shape. Brain pH was correlated with neuronal size, measured with both methods, and with neuronal shape. Age and post-mortem interval showed no correlations with any parameter. These data confirm the value of NeuN as a tool for quantitative neuronal morphometric studies in routinely processed human brain tissue. Absolute values are highly correlated between NeuN and cresyl violet stains, but cannot be interchanged. NeuN may be particularly useful when it is important to distinguish small neurons from glia, such as in cytoarchitectural studies of the cerebral cortex in depression and schizophrenia. PMID- 15130706 TI - Attentional demand for regulating postural sway: the effect of expertise in gymnastics. AB - A dual-task paradigm was used to investigate whether the expertise in motor skills requiring a fine postural control can modify the attentional demand necessary for regulating postural sway. Seven expert gymnasts and seven experts in other non-gymnastic sports were asked to respond as rapidly as possible to an unpredictable auditory stimulus while maintaining stable seated and in three upright postures of increasing difficulty: bipedal, unipedal, and unipedal on an unstable support (i.e. a 7 cm thick foam surface). RT values were used as an index of the attentional demand necessary for performing the postural tasks. Results showed that the attentional demand necessary for regulating postural sway increased as the postural task increased in difficulty. Interestingly, this effect was smaller for the gymnasts during unipedal stance. These findings suggest a decreased dependency on attentional processes for regulating postural sway during unipedal stance in gymnasts with respect to non-gymnasts. PMID- 15130707 TI - Kainic acid does not affect CA3 hippocampal region pyramidal cells in hypothyroid rats. AB - Thyroid hormones exert a crucial role on trophic events of the central nervous system during development, adulthood, and ageing. The deficiency of thyroid hormones could also produce a deficiency in neurotransmission in the hippocampal region. Kainic acid (KA) has become an important tool for studying functions related to excitatory amino acid transmission in mammals. Its neurotoxic effects on the pyramidal neurons of the CA3 hippocampal region are well known. We have examined the neurotoxicity of KA on these cells in hypothyroid rats. The hypothyroid state was induced by administration of methimazole. After 4 weeks of treatment, KA was administered once intraperitoneally at doses of 0, 1, 2.5, and 5mg/kg to the hypothyroid group, and 0 and 5mg/kg to the euthyroid group. In the euthyroid group, KA reduced the neuronal density in the CA3 hippocampal region, and in the hypothyroid rats with no administration of KA, the neuronal density of the CA3 hippocampal region is reduced also. Administering KA in hypothyroid rats did not reduce the number of CA3 pyramidal cells. PMID- 15130708 TI - Recovery from brain injury in animals: relative efficacy of environmental enrichment, physical exercise or formal training (1990-2002). AB - In the 1960s, it was shown for the first time that enriched housing enhances functional recovery after brain damage. During the 1970s and 1980s, many findings similar to this initial one have been reported, enlarging greatly its generality. Over the last 13 years, many different kinds of brain damage were modelled in animals or even directly studied in humans. Overall, these recent studies corroborated earlier findings, although occasional exceptions were reported. Other critical data, obtained mainly in intact animals, showed that enriched housing increases neurogenesis in the adult hippocampus. Recent evidence that this neurogenesis is involved in hippocampal-dependent learning supports the original interpretation of the enrichment effects as being the result of an accumulation of informal learning experiences (e.g., [. Heredity, environment, brain biochemistry, and learning. In: Current Trends in Psychological Theory. University of Pittsburgh Press, Pittsburgh, pp. 87-110;. Brain changes in response to experience. Sci. Am. 226, 22-29]). Other components of enriched environment, such as physical exercise, may have additive effects with those of training. The comparison of the relative effectiveness of enriched experience, of physical exercise and of training on structural and/or functional assessments of recovery, shows that training/learning is generally more effective than physical exercise and that enriched experience is a more potent therapy than either of these two other treatments. The combination of enriched experience with some other neurosurgical and/or neuropharmacological treatments may further improve its therapeutic effectiveness. Finally, other recent reports emphasize that the treatment parameters may be changed in order to approximate clinical/rehabilitation conditions and, nevertheless, remain effective. PMID- 15130709 TI - Signal transduction cascades underlying de novo protein synthesis required for neuronal morphogenesis in differentiating neurons. AB - Differentiating neurons must acquire many unique morphological and functional characteristics in creating the precise neural circuits of the mature nervous system. The phenomenon of 'neuronal differentiation' includes a special set of simple, separate processes, that is, neuritogenesis, neurite outgrowth, pathfinding, targeting and synaptogenesis. All of these processes are critically dependent on the reorganization of actin cytoskeleton by many actin-binding proteins that function downstream of Rho-family GTPases. Furthermore, de novo synthesis of key proteins are critically involved in the reorganization of actin cytoskeleton during neuronal differentiation. In this article, we review recent progresses in the general mechanisms that control actin dynamics by various actin binding proteins in differentiating neurons, including a series of recent studies from our laboratory on de novo synthesis of several key proteins that are essential for actin reorganization induced by second messengers. We demonstrated that dual regulation of cyclic AMP and Ca2+ determines cofilin (an actin-binding protein) phosphorylation states and LIM kinase 1 (a cofilin kinase) expression level during neuritogenesis. PMID- 15130710 TI - Pathophysiology of obsessive-compulsive disorder: a necessary link between phenomenology, neuropsychology, imagery and physiology. AB - Obsessive-compulsive disorder (OCD) is characterized by repetitive intrusive thoughts and compulsive time-consuming behaviors classified into three to five distinct symptom dimensions including: (1) aggressive/somatic obsessions with checking compulsions; (2) contamination concerns with washing compulsions; (3) symmetry obsessions with counting/ordering compulsions; (4) hoarding obsessions with collecting compulsions; and (5) sexual/religious concerns. Phenomenologically, OCD could be thought of as the irruption of internal signals centered on the erroneous perception that "something is wrong" in a specific situation. This generates severe anxiety, leading to recurrent behaviors aimed at reducing the emotional tension. In this paper, we examine how the abnormalities in brain activity reported in OCD can be interpreted in the light of physiology after consideration of various approaches (phenomenology, neuropsychology, neuroimmunology and neuroimagery) that contribute to proposing the central role of several cortical and subcortical regions, especially the orbitofrontal cortex (OFC), the anterior cingulate cortex (ACC), the dorsolateral prefrontal cortex (DLPC), the head of the caudate nucleus and the thalamus. The OFC is involved in the significance attributed to the consequences of action, thereby subserving decision-making, whereas the ACC is particularly activated in situations in which there are conflicting options and a high likelihood of making an error. The DLPC plays a critical part in the cognitive processing of relevant information. This cortical information is then integrated by the caudate nucleus, which controls behavioral programs. A dysfunction of these networks at one or several stages will result in the emergence and maintenance of repetitive thoughts and characteristic OCD behavior. PMID- 15130711 TI - [Non verbal communication abilities in severe traumatic brain injury]. AB - OBJECTIVE: Discursive abilities of severe brain injured patient are always impaired: loss of flexibility, lack of cohesion and coherence, often more elliptic. We know few about nonverbal competencies during discourse. The objective is to verify nonverbal abilities of these patients by pragmatic analysis. METHODS: Four men were examined more than 7 years after severe traumatic brain injury. Nonverbal Prutting and Kirchner Pragmatic Protocol (1987) were done allowing to a qualitative and quantitative measurement of paralinguistic behaviour: prosody and quality of speech, facial expression, posture, gaze, gesture. Two conditions were recorded: dual (descriptive discourse) and group (conversational discourse). Associated impairments such as cognitive and dysexecutive functioning were also investigated. RESULTS/DISCUSSION: Impoverishment (loss of ability) or impaired inadequacity was observed in all patients. Paralinguistic competences of conversational discourse was worse than descriptive one. Facial expression, gaze functioning, referential gesture were more often impaired. Maladjustment could be interpretated in reference with dysexecutive syndrome. CONCLUSION: In spite of the lack of information about the range of normal pragmatic behaviour, it seems that brain injured patients have shown poor nonverbal abilities during discourse. Rehabilitation training of communication skills would integrate this fact in order to improve interactivity and social relationship. PMID- 15130712 TI - [French version of structured interviews for the Glasgow Outcome Scale: guidelines and first studies of validation]. AB - OBJECTIVES: The Glasgow Outcome Scale (GOS) is the most widely used outcome measure after traumatic brain injury. The GOS's reliability is improved by a structured interview. The two aims of this paper were to present a French version of the structured interview for the five-point Glasgow Outcome Scale and the extended eight-point GOS (GOSE) and to study their validity. METHODS: The French version was developed using back-translation. Concurrent validity was studied by comparison with GOS/GOSE without structured interview. Inter-rater reliability was studied by comparison between assignments made by untrained head injury observers and trained head injury observers. Strength of agreement between ratings was assessed using the Kappa statistic. RESULTS: The French version and the guidelines for their use are given in the Appendix. Ratings were made for 25 brain injured patients and 25 relatives. Concurrent validity was good and inter rater reliability was excellent. CONCLUSION: Using the structured interview for the GOS will give a more reliable assessment of the outcome of brain injured patients by French-speaking rehabilitation teams and a more precise assessment with the extended GOS. PMID- 15130713 TI - [Influence of physical activity on postural capacities of elderly: effect of time of training]. AB - OBJECTIVE: The aim of this work was to define the influence of physical activity on static postural control of two groups of elderly people qualified as "non active" for the first one and "active" as far as the second group is concerned. The first group practised a physical activity during 3 months whereas the second group practised during 9 months. SUBJECTS AND METHOD: The two groups were composed of 21 subjects of 73 years-old and 10 subjects of 74 years-old. They were characterised by stabilometric data of surface, lengths and spontaneous sway in the lateral (X) and antero-posterior (Y) axis. The evaluations were separated by a period of 12 months and realised with open eyes and closed eyes on firm floor. RESULTS: The statistical analysis showed a significant difference of both groups on surface data in closed eyes condition (P < 0.04). CONCLUSION: The methodological conditions of the study and the heterogeneity of this kind of population could explain the lack of significant effect of the protocol. Data showed different postural adaptations between the two groups after training for a part of stabilometric parameters. Those data focused on the interest of the individualisation of training for elderly people. This approach may well favour the well-being and functional autonomy of this population. PMID- 15130714 TI - [The effect of similar speed's walking and functional classification of foot contact on variability of the vertical ground reaction force]. AB - INTRODUCTION: The variability of the normal ground reaction force (F(z)) is a restrictive factor for the clinical analyse of kinetics parameters recorded during walking. OBJECTIVE: The aim of this study is the decrease of the inter individual variability of the normal ground reaction force: F(z). MATERIAL AND METHOD: The method tested consists in imposing a similar speed to the subjects during walking tests, then to class foot according to their function: loading or propulsive foot. A group of seven young adults walk at spontaneous speed (VSpon) for the first walking test then at similar speed (Vsim = N(Fr) x square root of (g x li); where N(Fr) is the Froud's number, g is the gravitational acceleration and li is the length of the lower limb) for the second walking test. Two forces platforms register the F(z) of two consecutive steps. The normal ground reaction force and the mean coefficient of variability are retained to test for speed (VSim and VSpon) and functional classification effects. RESULTS: The CoV of F(z) decreases from 13% to 8% when the subjects walk at similar speed rather than at spontaneously chosen speed. The variability decreases by 1-1.5% when the data are classified according to functional criterion. The inter-individual variability of the F(z) significantly reduces when the tests are performed at similar speed and when the loading or propulsive factors are used for functional classification. CONCLUSION: The coupling of the methods described must permit to the clinician to constitute a data base which, tainted of less variability, should make easier the detection of pathology affecting the ground reaction force. PMID- 15130715 TI - [Interest of visual biofeedback training in rehabilitation of balance after stroke]. AB - OBJECTIVE: The aim of this study was the assessment of visual biofeedback in the management of balance disorders after stroke. MATERIAL AND METHODS: This is a prospective randomised trial, which compares visual biofeedback training and traditional balance training. Twenty-seven patients took part in this study: 13 in the first group and 14 in the second group. The rehabilitation program in the first group consists in an association of visual biofeedback and traditional training during 3 weeks. For the second group traditional training was used alone. Clinical, functional and forceplate assessments were performed before and after the program of rehabilitation. RESULTS: All the patients had a clinical and functional improvement. However, patients who have benefited from biofeedback training had a significant improvement of the forceplate parameters with closed eyes. DISCUSSION AND CONCLUSION: Those results confirm the interest of rehabilitation in balance disorders after stroke. The use of biofeedback training incorporated into functional physiotherapy affords added benefits probably by a best integration of proprioceptive informations. PMID- 15130717 TI - [Chronicity, recurrence, and return to work in low back pain: common prognostic factors]. AB - OBJECTIVE: To determine the common risk factors of recurrence, chronicity and non return to work in low back pain. MATERIAL AND METHOD: A systematic review of the literature was done by searches of Medline, Embase, Pascal, the Cochrane database and also in the unindexed literature. Keywords used were low back pain, chronic, risk factors, recurrence, predictive value of tests, prognosis, confounding factors. Studies were assessed by two readers using the ANAES (French Agency for Health Assessment) scale allowing classification into high-, moderate- and low quality trials. The scientific evidence level of the identified risk factors depend on the methodological quality of the studies, the number of studies in agreement, the coherence of their results and their clinical relevance. RESULTS: Fifty-four high quality studies were included. Several prognostic factors are common to the three described clinical situations. A history of low back pain (including the concept of pain severity, duration, disability, leg pain, related sickness leave and a history of spinal surgery), low level of job satisfaction and poor general health are highlighted with a strong level of evidence. Socioprofessionel and psychological factors including employment status, amount of wage, workers' compensation, and depression were found but with moderate level of evidence. Physical factors including lifting time per day and work postures were also found with moderate level of evidence. CONCLUSION: This study confirm that several prognostic factors are commun to recurrence, chronicity and non return to work in low back pain. Early identification of these factors is important in understanding, and hopefully preventing, the recurrence or the progression to chronicity and disability in low back trouble. PMID- 15130718 TI - Molecular responses of vascular smooth muscle cells and phagocytes to curcumin eluting bioresorbable stent materials. AB - A major complication of coronary stenting is restenosis, often accompanied by inflammatory reactions and smooth muscle cell proliferation. Curcumin has been shown to possess anti-inflammatory and anti-proliferative properties, thus we hypothesize that locally released curcumin by coronary stent would diminish in stent restenosis. As a first test of this hypothesis, curcumin-eluting PLLA films (C-PLLA) were produced and the anti-inflammatory and anti-proliferative properties were then tested using peritoneal phagocytes and human coronary artery smooth muscle cell (hCASMCs) culture systems. We find that the addition of curcumin reduced phagocyte accumulation and activation on C-PLLA films. On the other hand, C-PLLA significantly reduced the proliferation, but not the adhesion, of hCASMCs. The molecular responses of hCASMCs to C-PLLA were further assessed by cDNA microarray analysis. Curcumin up-regulated genes related to apoptosis and enhanced the expression of anti-proliferative and anti-inflammatory factors, and of antioxidants. Equally important, C-PLLA inhibited the cell cycle progression of adherent hCASMCs. The results suggest that curcumin regulates gene expression and cell function through the protein kinase (PK) and mitogen-activated protein kinase (MAPK) pathways. These results support the use of curcumin to inhibit in stent restenosis. PMID- 15130719 TI - Investigation of the activation of a human serum complement protein, C3, by orthopedic prosthetic particulates. AB - Myriad molecular, cellular, and physiological processes underlie the inflammatory and osteolytic processes induced by particles of biomaterials resulting from the wear of implants such as total joint replacement prostheses. The objective this study was to investigate the role that the complement system may be playing in these phenomena. The aim was to evaluate the degree to which particles of selected orthopaedic materials--high density and ultrahigh molecular weight polyethylene, polymethylmethacrylate, and commercially pure titanium--cause the elevation of a key complement molecule, C3a, in an in vitro assay that directly measured the concentration of C3a. The results demonstrated that HDPE particles, at high concentration, are capable of causing the elevation of C3a in the in vitro assay. This finding is discussed in the context of other work and the mechanics of the complement system as it may affect the osteolytic process. PMID- 15130720 TI - Nano-scale surface modification of a segmented polyurethane with a phospholipid polymer. AB - Nano-scale modification of a segmented polyurethane (SPU) with cross-linked 2 methacryloyloxyethyl phosphorylcholine (MPC) polymer was performed to obtain a biocompatible elastomer. To control the domain size and the depth of the modified layer, various compositions of monomers, including MPC, 2-ethylhexyl methacrylate (EHMA), and glycerol 1,3-diglycerolate diacrylate, were examined. SPU film was immersed in the monomer solution and visible light irradiation was applied to initiate polymerization to the SPU film that was held by mica to condense MPC units at the surface. The surfaces of the obtained film were analyzed by X-ray photoelectron spectroscopy and water contact angle measurement. The surface density of MPC units changed with the monomer concentration, and the density was the highest when the ratio between MPC and EHMA was 7:3. In modified SPU films, 6 to 25-nm MPC unit-enriched domains were observed and the density of these domains gradually decreased with depth. The sizes of the domains depended on the MPC composition in the monomer solution. The mechanical properties of the modified films as evaluated by tensile strength measurement under wet conditions were not significantly different from those of SPU. With increase in the existence of MPC unit-enriched domains on the MEG film surface, platelet adhesion and activation were remarkably reduced compared to the SPU film. This nano-scale surface modification may be a useful technique for applying elastic polymer biomaterials. PMID- 15130721 TI - Influence of interphase layer on the overall elasto-plastic behaviors of HA/PEEK biocomposite. AB - A three-dimensional finite element unit cell model has been designed and constructed for studying mechanical properties of hydroxyapatite (HA) reinforced polyetheretherketone (PEEK) biocomposite. The model consists of an elastic brittle HA spherical particle, an elasto-plastic matrix and an interphase layer between the particle and the matrix. The interphase layers with four different kinds of material behaviors have been taken into consideration to examine their effects on the overall properties of the composite. The damage evolution in the matrix and the interphase layer, and the interface failure, were also taken into account. Some other factors, such as mesh sensitivity, loading velocity and mass scale scheme, were also discussed in this investigation. A general-purpose finite element software package, ABAQUS, incorporated with a user-defined material subroutine, was used to perform the analysis. The predicted results were compared with the experimental data obtained from existing literatures. The results predicted by using the cell model with consideration of the matrix degradation and the effects of the damage and failure on the interphase layer are in good agreement with the experimental ones. Hence, the suitability of our proposed cell model incorporated with an appropriate type of the interphase layer for modeling the mechanical properties of the particulate biocomposite could be verified. PMID- 15130722 TI - Stimulation of porcine bone marrow stromal cells by hyaluronan, dexamethasone and rhBMP-2. AB - In the interest of optimizing osteogenesis in in vitro, the present study sought to determine how porcine bone marrow stromal cell (BMSc) would respond to different concentrations of hyaluronan (HY) and its different combinations with dexamethasone (Dex) and recombinant human bone morphogenic protein-2 (rhBMP-2). Cellular proliferation was determined by 3H-thymidine incorporation into DNA at both Days 2 and 7 when BMSc was cultivated with HY at concentrations of 0, 0.5, 1.0, 2.0 and 4.0 mg/ml. HY accelerated cellular proliferation when compared with cultures in the absence of HY at both Days 2 and 7. BMSc proliferation under the high HY concentration of 4 mg/ml was significantly higher than under the other, lower HY concentrations of 0.5, 1.0 and 2.0 mg/ml. When BMSc were cultivated under HY at concentrations of 0, 1.0 and 4.0 mg/ml and its 12 combinations with rhBMP-2 at concentrations of 0 and 10 ng/ml and Dex (+, -) at both Days 2 and 7, cellular responses were examined by 3H-thymidine incorporation into DNA, cellular alkaline phosphatase (ALP) activity, and pro-collagen type I C-terminal propeptide production. HY accelerated cellular proliferation irrespective of the presence of Dex and rhBMP-2. HY increased expression of ALP activity at Day 7, whereas had inhibitory effect at Day 2. HY and Dex showed an interaction on expression of ALP acitivity irrespective of the HY dose by Day 7. Collagen synthesis was inhibited by HY irrespective of the presence of other factors at both Days 2 and 7. When BMSc were cultivated with HY of 4.0 mg/ml alone, its combinations with Dex (+) and 10 ng/ml rhBMP-2, and with DMEM/FBS alone, expression of bone-related marker genes was evaluated by real-time reverse transcription-polymerase chain reaction (Real-time RT-PCR) analysis. Osteocalcin was up-regulated under both rhBMP-2 and HY-Dex-rhBMP-2 at Day 2, as also under 4 mg/ml HY, Dex, HY-Dex, Dex-rhBMP-2, and HY-Dex-rhBMP-2 by Day 7. Type 1alpha1 collagen was induced by rhBMP-2 on Day 2, and by Dex-rhBMP-2 on Day 7. Osteonectin and type X collagen was only marginally induced by HY at Day 2. Type 1alpha1 collagen and type X collagen were down-regulated in the presence of 4 mg/ml HY by Day 7. These results suggest that HY stimulates BMSc proliferation, osteocalcin gene expression, and a secretion of enzymes such as that of ALP activity in vitro. More importantly, HY can interact with Dex and rhBMP-2 to generate direct and specific cellular effects, which could be of major importance in bone tissue engineering. PMID- 15130723 TI - De novo bone formation using bovine collagen and platelet-rich plasma. AB - In order to regenerate critical-size bone defects, a variety of bone substitutes is used in addition to autogenous bone. The regenerative capacity of these bone substitutes is usually compared to the efficacy of autogenous bone known as the "golden standard". Different cytokines influence the regeneration process because of their morphogenic or mitogenic properties. Platelet-rich plasma (PRP), a platelet concentrate, is characterised by having a positive effect on wound healing, reducing bone graft resorption and increasing the density of bone transplants. This experiment was commenced with a view to studying the osseous defect regeneration after placing various combinations of "filler materials" in experimentally created defects in the forehead of adult pigs. Regeneration by means of grafted autogenous bone (Group 1) or a bovine collagen based medical device (Group 4) alone and combined with PRP in two concentrations (Groups 2, 3, 5 and 6) was evaluated by means of microradiography and light microscopy after 2, 4 and 12 weeks. The microradiographic and light microscopic findings showed that autogenous bone in combination with PRP (Groups 2 and 3) had a significant accelerating effect on early bone regeneration (2 weeks). This effect was not evident when PRP was added to the bovine collagen (Groups 5 and 6). When using the collagen alone, significantly higher mineralisation values were achieved after 2 and 4 weeks than when using autogenous bone alone. After a 12-week observation period, the existing differences between the healing processes in the various groups were more or less levelled out. In summary, the results of the study indicate that clinically autogenous bone, as expected, is the ideal defect filler. Combining autogenous bone with PRP did not provide significantly better results. The findings in the groups treated with bovine collagen indicate that its local application mimics the effect of autogenous bone and amplifies bone regeneration when comparing with the control defect. PMID- 15130724 TI - Electrochemically assisted co-precipitation of protein with calcium phosphate coatings on titanium alloy. AB - A bovine serum albumin protein-containing calcium phosphate coating (BSA/brushite) was prepared by electrochemically assisted co-precipitation onto a hydroxyapatite (HA) coated Ti-6Al-4V surface. Electrochemically assisted co precipitation of BSA/brushite coatings onto HA resulted in a 70-fold increase in BSA inclusion compared to simple adsorption, and was subsequently released by a slower mechanism (15% loss over 70 h). Thus, this electrochemically assisted co precipitation technique provides an efficient method of protein incorporation at physiological temperature, with a potential for sustained release of therapeutic agents as may be required for metallic implant fixation. PMID- 15130725 TI - Cell response to dextran-derivatised iron oxide nanoparticles post internalisation. AB - Magnetic nanoparticles have been used for many years as magnetic resonance imaging contrast agents. Despite the fact that there are currently several dextran-coated iron oxide nanoparticles are in preclinical and clinical use, there is very little information available concerning the influence such particles have on cells in culture. The prerequisite for particles employed as contrast agents is capture and subsequent uptake by cells. This study involved the use of magnetic nanoparticles synthesised and derivatised with dextran, compared to similar underivatised plain particles. The influence in vitro was assessed using human dermal fibroblasts and various techniques to observe cell particles interaction, including light and fluorescence microscopy, scanning and transmission electron microscopy. The results indicate that although both the uncoated and the dextran-derivatised particles are uptaken into the cell, the derivatised particles induce alterations in cell behaviour and morphology distinct from the plain particles, suggesting that cell response is dependent on the particles coating. PMID- 15130726 TI - Changes in fibroblast morphology in response to nano-columns produced by colloidal lithography. AB - In designing new biomaterials, specific chemical and topographical cues will be important in guiding cell response. Filopodia are actin-driven structures produced by cells and speculated to be involved in cell sensing of the three dimensional environment. This report quantifies filopodia response to cylindrical nano-columns (100 nm diameter, 160 nm high) produced by colloidal lithography. Also observed were actin cytoskeleton morphology by fluorescence microscopy and filopodia morphology by electron microscopy (scanning and transmission). The results showed that the fibroblasts used produced more filopodia per microm of cell perimeter and that filopodia could often be seen to interact with the cells' nano-environment. By understanding as to which features evoke spatial reactions in cells, it may be possible to design better biomaterials. PMID- 15130727 TI - Chlorhexidine release from an experimental glass ionomer cement. AB - Glass ionomer cements (GIC) can potentially be used as matrices for the slow release of active species, as has been shown previously for fluoride ions. This study investigated the use of an experimental GIC as a carrier for the release of chlorhexidine acetate (CHA) at included concentrations ranging from 0.5% to 13.0% of CHA by weight. Release into water was examined using high-performance liquid chromatography. All measurable chlorhexidine was released within 22 h1/2, however this was less than 10% of the total mass incorporated in the specimens. An increased percentage of CHA incorporated into the powder gave an increased release into the surrounding water. The bulk of the CHA was retained within the cement. For comparison, the surface chemistry of a CHA-containing GIC was examined using X-ray photoelectron spectroscopy before and after prolonged immersion in water. This confirmed retention of a large amount of CHA. Spectra after leaching appeared very similar to those from a CHA-free GIC after immersion in a CHA solution. In order to explore the effect of CHA-inclusion on the cement properties, compressive strengths, working and setting times were also measured. In general, compressive strengths were found to be decreased in direct proportion to the quantity of CHA added, while working and setting times increased. PMID- 15130728 TI - In vivo studies of endotoxin removal by lysine-cellulose adsorbents. AB - A new type adsorbent for removal of bacterial endotoxins was prepared by immobilizing lysine covalently onto cellulose beads. Endotoxins (Escherichia coli O55: B5) were injected into 13 healthy New Zealand white rabbits to induce infectious symptoms. Hemoperfusion using the adsorbent column removed endotoxins in the blood of eight rabbits during 2h while other five rabbits were used as control. The mean blood endotoxin concentration was reduced significantly from 5.56 +/- 0.54 EU/ml (1 EU = 100 pg) before treatment to 0.41 +/- 0.26 EU/ml after perfusion as measured by the limulus amebocyte lysate test (Chromogenix). Liver function and renal function tests showed significant improvement of septic symptoms in contrast to the control group. Other parameters such as superoxide dismutase and malondialdehyde were ameliorated markedly after the treatment. Moreover, the adsorbent showed good results in mechanical strength, blood compatibility and cytotoxicity, which suggested that lysine-cellulose adsorbent was of high ET-binding efficacy without significant side effect. It has a high potential of clinical application for treatment of patients with severe sepsis. PMID- 15130729 TI - The pKa effects of the carboxylic acid in N-methacryloyl-omega-amino acid on the demineralization and bond strengths to the teeth. AB - It is understood that the application of a self-etching primer to the tooth enhances the bonding of the resin to the tooth. In this study, we designed a self etching primer consisting of a series of three N-methacryloyl-omega-amino acids (NMomegaA) with different methylene chain lengths. The demineralization aspect of the teeth components by the carboxylic acid in the NMomegaA and its effects on the bond strength of the resin to the tooth were examined. The amount of decalcification of the hydroxyapatite or dentin by the carboxylic acid in the NMomegaA was strongly dependent on the carboxylic acid's pKa value in the NMomegaA. However, the bond strength's mean values for both the enamel and dentin were not influenced by the degree of demineralization by the carboxylic acid in the NMomegaA. The greater mean value of the dentin's bond strength than with the enamel's was due to differences in the adhesion mechanism types, since the NMomegaA not only exhibited an etching efficacy but also a priming efficacy to the collagen that had been exposed by the NMomegaA conditioning. PMID- 15130730 TI - Positron emission tomography: an advanced nuclear medicine imaging technique from research to clinical practice. PMID- 15130731 TI - Biophysical basis of magnetic resonance imaging of small animals. PMID- 15130732 TI - Nuclear magnetic resonance in laboratory animals. PMID- 15130733 TI - Simultaneous electrophysiology and functional magnetic resonance imaging studies in conscious rats. PMID- 15130734 TI - Functional magnetic resonance imaging of macaque monkeys. PMID- 15130735 TI - Atlas template images for nonhuman primate neuroimaging: baboon and macaque. PMID- 15130736 TI - Direct comparison of visual cortex activation in human and nonhuman primates using functional magnetic resonance imaging. PMID- 15130737 TI - Magnetic resonance imaging of brain function. PMID- 15130738 TI - Laser-polarized xenon nuclear magnetic resonance, a potential tool for brain perfusion imaging: measurement of the xenon T1 in vivo. PMID- 15130739 TI - Positron emission tomography receptor assay with multiple ligand concentrations: an equilibrium approach. PMID- 15130740 TI - Estimation of local receptor density, B'max, and other parameters via multiple injection positron emission tomography experiments. PMID- 15130741 TI - Imaging dopamine receptors in the rat striatum with the MicroPET R4: kinetic analysis of [11C]raclopride binding using graphical methods. PMID- 15130742 TI - Quantitative analysis of dopamine D2 receptor kinetics with small animal positron emission tomography. PMID- 15130743 TI - Magnetic resonance imaging in biomedical research: imaging of drugs and drug effects. PMID- 15130744 TI - Enzyme-dependent fluorescence recovery after photobleaching of NADH: in vivo and in vitro applications to the study of enzyme kinetics. PMID- 15130745 TI - Imaging myocardium enzymatic pathways with carbon-11 radiotracers. PMID- 15130746 TI - Molecular imaging of enzyme function in lungs. PMID- 15130747 TI - Brain uptake and biodistribution of [11C]toluene in nonhuman primates and mice. PMID- 15130748 TI - Optimizing luciferase protein fragment complementation for bioluminescent imaging of protein-protein interactions in live cells and animals. PMID- 15130749 TI - Mitochondrial NADH redox state, monitoring discovery and deployment in tissue. PMID- 15130751 TI - Effects of simultaneous over-expression of Cu/ZnSOD and MnSOD on Drosophila melanogaster life span. AB - The FLP-out technique, based on yeast FLP recombinase, allows induced over expression of transgenes in Drosophila adults. With FLP-out control and over expressing flies have identical genetic backgrounds and therefore differences in life span must result from transgene induction. The amount of over-expression achieved varies between independent transgenic lines, and previously for both Cu/ZnSOD and MnSOD life span was found to be increased in proportion to the increase in enzyme activity. To determine if greater increases in enzyme and life span could be achieved with FLP-out, enzyme over-expression and life span were analyzed in eight lines containing two MnSOD transgenes, three lines containing three MnSOD transgenes, and three lines containing a MnSOD transgene plus a Cu/ZnSOD transgene. Life span was again found to be increased in proportion to the increase in MnSOD enzyme activity, with increases of up to 40% in mean and maximum life span. However the increases in enzyme activity and life span conferred per transgene were reduced when more than one transgene was present at the same time. When the reduced efficiency of enzyme over-expression per transgene was taken into account, simultaneous over-expression of MnSOD and Cu/ZnSOD was found to have partially additive effects on life span. PMID- 15130752 TI - Histone deacetylases, HDAC1 and HSIR2, act as a negative regulator of ageing through p53 in human gingival fibroblast. AB - Histone deacetylases (HDACs) such as HDAC1 and HSIR2 have been known to be involved in the regulation of life-span extension. However, its underlying mechanism remains unclear in human. Using the primary human gingival fibroblasts (HGFs) derived from donors of different ages, which exhibit clear features of senescence in aged HGFs, we demonstrated that histone deacetylase, HDAC1 and HSIR2, repressed the ageing through the transcriptional inactivation of p53 and p21 promoters. These results suggest that primary HGFs can be a useful human ageing model, and HDAC1, HSIR2, p53 and p21 may play an important role in ageing process of human beings. PMID- 15130753 TI - Selective COX-2 inhibitor, NS-398, inhibits the replicative senescence of cultured dermal fibroblasts. AB - Cyclooxygenase 2 (COX-2) is known to be increased in aged cells. Recent studies suggest that the increased expression of COX-2 may be involved in the pathogenesis of age-associated diseases such as rheumatoid arthritis and cancer. We investigated the role of COX-2 in cell cycle arrest and collagen deficiency during the aging process. Using the replicative senescence model of dermal fibroblasts, we demonstrated the increased expression of COX-2 and increased PGE(2) levels associated with replicative senescence. Replicative senescent cells showed a decreased ability to induce cell proliferation, probably due to the increased expression of the p53 protein and the decreased expression of the PCNA protein, and also showed increased expression of MMP-1, and decreased expression of tissue inhibitor of metalloproteinase-1 (TIMP-1) and procollagen. The selective COX-2 inhibitor, NS-398, can inhibit the senescence-associated increases of COX-2, PGE(2), p53 and MMP-1 expression, and the senescence associated decreases of PCNA, TIMP-1 and procollagen expression. These results suggest that the increased level of COX-2 and higher level of PGE(2) in aged cells may play an important role in cellular senescence, and that selective COX-2 inhibitors may be useful for the intervention of skin aging. PMID- 15130754 TI - Differential variation of mitochondrial H2O2 release during aging in oxidative and glycolytic muscles in rats. AB - Mitochondrial free radical (ROS) production could be involved in sarcopenia. Our aim was to measure this production in various muscles during aging. Male Wistar rats aged 4.5 and 24 months were used. H(2)O(2) release and protein carbonyls were evaluated in isolated mitochondria from an oxidative (soleus) and a glycolytic (tibialis anterior) muscle. Total and Mn-superoxide dismutase (SOD), catalase, glutathione peroxidase (GPX) and glutathione reductase (GR) activities were measured in tibialis anterior. In soleus, glutamate/malate supported mitochondrial H(2)O(2) release was lower than in tibialis anterior in young rats, but increased significantly with age. In tibialis anterior, glutamate/malate or succinate supported H(2)O(2) release was unchanged with age. ROS generators were complexes I and III. Mitochondrial carbonyl content remained stable during aging in both muscles but tended to be higher in tibialis anterior than in soleus. Tibialis anterior total SOD (+17%), catalase (+84%), and GPX (-17%) activities varied significantly with age but Mn-SOD was unchanged, suggesting an increase in cytosolic ROS production. In conclusion, the higher life-long H(2)O(2) release observed in tibialis anterior is consistent with the known sensitivity of glycolytic muscles to sarcopenia. The fact that the rate of H(2)O(2) release increases with age in soleus seems to have little impact. PMID- 15130755 TI - Dietary restriction: effects of short-term fasting on protein uptake and cell death/proliferation in the rat liver. AB - Dietary restriction (DR) is known to prolong life in laboratory animals. Intermittent (alternate-day) fasting or short-term repeated fasting has also been reported to increase the life span of animals. In the present study, we investigated the changes or induction of abnormalities of protein metabolism in rats during fasting, and measured asialoglycoprotein uptake and cell death/proliferation in the liver of rats receiving fasting and refeeding. In the results, liver weight decreased significantly after 48 h of fasting and increased during the refeeding period, returning to the pre-fasting level by 12 h of refeeding. Cell death, determined by single stranded DNA (ssDNA) staining method, increased during the fasting period, and returned to the pre-fasting level during the refeeding period. Cell proliferation, determined using antibodies (Ab) against proliferating cell nuclear antigen, decreased during the fasting period, and increased during the refeeding period. Changes in cell death and cell proliferation were inversely related. However, there was no significant difference in asialoglycoprotein uptake by the whole liver between the ad libitum (AL)-fed rats and 48 h fasted rats. Thus, neither the changes in liver weight nor cell death/proliferation affected asialoglycoprotein uptake on a living body. These results suggest that episodes of 48 h fasting do not induce protein metabolism abnormalities in the liver. PMID- 15130756 TI - Body weight, hormones and T cell subsets as predictors of life span in genetically heterogeneous mice. AB - Previous studies have shown that T cell subset levels, early life body weight, and levels of leptin and thyroid hormones can each serve, independently, as predictors of life span in populations of genetically heterogeneous mice. New data now confirm, in a replicate cohort, that T cell subset patterns predict longevity, and show that they can do so when measured in mice as young as 8 months of age. Individual T cell subsets, as well as composite indices that combine data from two or more T cell measures at 8 or 18 months, can be combined with 3- and 9-month body weight data to provide better prediction of life span than either immune or weight measures alone. Mice whose immune and weight measures are both in the lowest quartile have mean and maximal life spans that are 18% and 16-25% higher, respectively, than mice in the opposite quartiles for both traits. Thyroxine levels measured at 4 months lead to further improvement over models that combine weight and immune data only. A genome scan provided evidence for loci on chromosomes 2, 12, 13, and 17 that modulate age-sensitive T cell subset patterns at both 8 and 18 months of age. These data show that late life mortality risks are influenced to a measurable degree by factors that modulate growth trajectory and hormone and immune status in the first third of the life span, and provide clues as to which early life systems deserve further scrutiny as potential mediators of late life disease risk. PMID- 15130757 TI - Modest implication of interleukin-6 promoter polymorphisms in longevity. AB - The multifunctional interleukin-6 has been suggested to contribute to a chronic low-grade inflammatory status, thereby conferring susceptibility to age-related pathological conditions as well as functional decline and increased mortality. Several polymorphisms have been identified in the interleukin-6 promoter, but investigation of the effect of these on interleukin-6 levels and disease susceptibility have led to contradictory results. This study investigates the significance of the three single-point polymorphisms (-597G/A, -572G/C and 174G/C) and the AT-stretch polymorphism (-373(A)n(T)m) in ageing, by comparison of the frequency of each single polymorphism separately as well as the entire promoter haplotype in a total of 1710 Danish subjects ranging in age from 47 to 100 years. We found a modest, but significant, increase in the frequency of interleukin-6 -174GG homozygotes with age suggesting that this genotype is advantageous for longevity. PMID- 15130758 TI - Roles of p38 and JNK mitogen-activated protein kinase pathways during cantharidin induced apoptosis in U937 cells. AB - Cantharidin is an active compound from blister beetles traditionally used for the treatment of cancer. It is known to exert its antitumor activity by inducing apoptosis in cancer cells. However, its signaling pathway still remains unclear. Therefore, we investigated the roles of the mitogen-activated protein kinases (MAPKs) and the tumor suppressor gene, p53, during cantharidin-induced apoptosis in U937 human leukemic cells. Cantharidin effectively activated ERK-1/2, p38 and JNK in U937 cells in a time- and dose-dependent manner. Cantharidin also exhibited a strong cytotoxicity and induced apoptosis in U937 cells. For the evaluation of the role of MAPKs, PD98059, SB202190 and SP600125 were used as MAPK inhibitors for ERK-1/2, p38 and JNK. PD98059 did not affect cantharidin-induced cytotoxicity and apoptosis, whereas SB202190 and SP600125 significantly interfered with cytotoxic and apoptotic activities induced by cantharidin. Cantharidin alone induced the apoptosis by phosphorylation of p53, up-regulation of downstream target genes, MDM2 and p21 and also cleaved caspase-3, whereas SB202190 and SP600125 caused the down-regulation of p53, MDM-2, p21 and cleaved caspase-3 after a co-treatment with cantharidin. Similarly, SB202190 and SP600125 significantly disturbed the caspase-3 activity after a co-treatment with cantharidin by colorimetric assay. Taken together, these results suggest that cantharidin can induce apoptosis by activation of p38 and JNK MAP kinase pathways associated with p53 and caspase-3. PMID- 15130759 TI - Tetrandrine-induced apoptosis is mediated by activation of caspases and PKC-delta in U937 cells. AB - Tetrandrine, which is isolated from Chinese herb Stephania tetrandrae, possesses anti-inflammatory, immunosuppressive, and cytoprotective properties. Though it was previously shown that tetrandrine causes a G1 blockade and apoptosis in various cell types, however, the mechanism by which tetrandrine initiates apoptosis remains poorly understood. In present study, we investigated the mechanisms of apoptosis induced by tetrandrine in U937 leukemia cells. Tetrandrine inhibited U937 cell growth by inducing apoptosis. After treatment of U937 cells with tetrandrine (10microM) for 24h, alteration of cell morphology, chromatin fragmentation, cytochrome c release, and caspase activation were observed. Tetrandrine also induced early oxidative stress, which resulted in activation of JNK, but not ERK and p38 MAPK. A broad-spectrum caspase inhibitor and antioxidants significantly blocked tetrandrine-induced caspase-3 activation. However, inhibition of the JNK activity with SP600125 did not block tetrandrine induced apoptosis. Tetrandrine-induced apoptosis of U937 cells also required activity of PKC-delta, because pretreatment with a specific PKC-delta inhibitor greatly blocked tetrandrine-induced caspase-3 activation. In addition, the apoptotic response to tetrandrine was significantly attenuated in dominant negative PKC-delta transfected MCF-7 cells, suggesting that PKC-delta plays an important role in tetrandrine-induced apoptosis and can induce caspase activation. These results suggest that tetrandrine induces oxidative stress, JNK activation, and caspase activation. However, JNK activation by ROS is not involved in the tetrandrine-induced apoptosis. In addition, tetrandrine induces caspase-dependent generation of a catalytically active fragment of PKC-delta, and this fragment also appears to play a role in the activation of caspases. PMID- 15130760 TI - Differential activation of CYP2C9 variants by dapsone. AB - Studies have shown that CYP2C9.1 mediated metabolism of flurbiprofen or naproxen is activated by co-incubation with dapsone. However, dapsone activation has not been examined in the known variant forms of CYP2C9. Six concentrations of flurbiprofen (2-300microM) or naproxen (10-1800 microM) were co-incubated with six concentrations of dapsone (0-100 microM) and with reconstituted, purified CYP2C9.1, CYP2C9.2 (R144C), CYP2C9.3 (I359L), or CYP2C9.5 (D360E), in order to assess degrees of activation. Dapsone increased the efficiency (V(m)/K(m)) of flurbiprofen 4'-hydroxylation by CYP2C9.1, CYP2C9.2, CYP2C9.3, and CYP2C9.5 by 8 , 31-, 47-, and 22-fold, respectively. In similar experiments using the substrate naproxen, dapsone increased the efficiency of naproxen demethylation 7-, 15-, 13 , and 22-fold, in CYP2C9.1, CYP2C9.2, CYP2C9.3, and CYP2C9.5, respectively. Also, dapsone normalized naproxen's kinetic profile from biphasic (CYP2C9.1 and CYP2C9.2) or linear (CYP2C9.3 and CYP2C9.5) to hyperbolic for all variant forms. Thus, amino acid substitutions of CYP2C9 variants affect the degree of dapsone activation in a genotype-dependent fashion. Furthermore, the degree of effect noted across variants appeared to be dependent on the substrate studied. PMID- 15130761 TI - Diazoxide affects the IF1 inhibitor protein binding to F1 sector of beef heart F0F1ATPsynthase. AB - Diazoxide, a selective opener of the mitochondrial ATP-sensitive K+ channel (mitoK(ATP)), has been reported to enhance F(0)F(1)ATPsynthase inhibition during ischemia, but the underlying mechanisms are still unclear. Here, we demonstrate that diazoxide directly interacts with the F(1) sector of beef heart F(0)F(1)ATPsynthase markedly promoting the binding of the inhibitor protein (IF(1)) to beta subunit. More specifically, the treatment of soluble F(1) with one equivalent of diazoxide was sufficient to decrease the K(d) of IF(1)-F(1) complex at low pH. Such effect was revealed only on the cycling enzyme, while no effect was observed in the absence of Mg-ATP. However, diazoxide binding occurred independently from the catalysis, as shown by the structural changes induced by the drug in not catalytically active F(1) and revealed by CD spectra. In addition, kinetic analysis of ATP hydrolysis demonstrated that diazoxide exerts a stabilising role on Mg-ADP bound in the catalytic site of the beta subunit adopting the tight conformation (beta(DP)). In accordance, a stabilising effect of Mg-ADP at the nucleotide binding domain (NBD) has been reported also for K(ATP) channel. These results suggest that diazoxide binds to beta subunit at NBD, which is highly conserved in the ATP-binding cassette protein family, thus inducing nucleotide stabilisation and favouring F(1) conformation suitable for IF(1) binding. Finally, diazoxide also increased IF(1) binding to membrane bound F(1), while it did not influence the energisation-dependent IF(1) release. As IF(1) binding mediates the F(0)F(1)ATPsynthase inhibition, we suggest that such mechanism may contribute to cardioprotection during ischemia. PMID- 15130762 TI - Human alpha1D-adrenoceptor phosphorylation and desensitization. AB - Rat-1 fibroblast were transfected with a plasmid containing the cDNA of the human alpha(1D)-adrenoceptor. A cell line was isolated that stably expressed the receptor as evidenced by BMY 7378-sensitive noradrenaline-induced increases in intracellular calcium concentration. The effect of noradrenaline was blocked by active phorbol esters; such blockade was mediated by protein kinase C (PKC) as evidenced by its inhibition by staurosporine or the downregulation of this protein kinase. Radioligand binding experiments showed expression of receptors with high affinity for [3H]tamsulosin (K(D) 0.30 +/- 0.05 nM) but low density (B(max) 35 +/- 4 fmol/mg protein). The receptors had the expected orders of potency for agonists (adrenaline = noradrenaline > oxymetazoline) and antagonists (BMY 7378 > 5-methyl-urapidil = phentolamine). Photoaffinity labeling identified the receptor as a band of M(r) 70-80kDa, which could be immunoprecipitated with a selective anti-alpha(1D)-adrenoceptor antiserum. In cells metabolically labeled with radioactive phosphate the adrenoceptor was identified as a phosphoprotein whose phosphorylation state was increased by the agonist, noradrenaline, and by phorbol myristate acetate. The data indicate that the human alpha(1D) adrenoceptor function was regulated through phosphorylation by PKC. PMID- 15130763 TI - Involvement of tyrosine kinase p56/Lck in apoptosis induction by anticancer drugs. AB - Induction of apoptosis is a hallmark of the cellular response of human lymphocytes and lymphoma cells to treatment with anticancer drugs and irradiation. Both treatment modalities trigger apoptosis through intrinsic, mitochondrial apoptosis pathways resulting in the activation of caspases. We and others have shown that the tyrosine kinase p56/Lck is involved in the regulation of apoptosis induced by irradiation or treatment with ceramide but dispensable for death receptor triggered cell death. However, the role of p56/Lck for apoptosis induction in response to anticancer drugs is unclear. To elucidate the putative requirement of p56/Lck for apoptosis signaling of cytotoxic drugs, activation of caspases and alteration of mitochondrial functions were determined in Jurkat T cells, the p56/Lck deficient JCaM1.6 cells and the p56/Lck retransfected JCaM1.6/Lck cells in response to chemotherapeutic drugs with different targets of their primary action. Treatment with Doxorubicin, Paclitaxel or 5-Fluorouracil induced a breakdown of the mitochondrial membrane potential and apoptotic cell death in p56/Lck expressing Jurkat and the retransfected JCaM1.6/Lck cells within 48h of treatment. However, almost no mitochondrial alterations and no induction of apoptosis could be detected in the p56/Lck deficient JCaM1.6 cells. Correspondingly, activation of caspases-9, -8, and -3 and cleavage of the caspase-3 substrate PARP (poly-(ADP-ribose)-polymerase) were almost completely absent in JCaM1.6 cells while present in p56/Lck positive Jurkat and JCaM1.6/Lck cells. In contrast, retransfection of the cells with the p56/Lck-related tyrosine kinase Src could not restore sensitivity to the treatment with cytotoxic drugs indicating a specific role of the tyrosine kinase p56/Lck in apoptosis signaling. Importantly, kinase-activity of p56/Lck may be dispensable for its pro-apoptoptic action since preincubation with the Src-kinase inhibitor PP2 did not reduce apoptosis induced by cytotoxic drugs. In conclusion, the tyrosine kinase p56/Lck is essential for apoptosis induction by Doxorubicin, Paclitaxel and 5-Fluorouracil regulating early steps of the mitochondrial apoptosis signaling cascade, including alteration of mitochondrial functions and caspase-activation. PMID- 15130764 TI - Cell cycle- and protein kinase C-specific effects of resiniferatoxin and resiniferonol 9,13,14-ortho-phenylacetate in intestinal epithelial cells. AB - We have previously reported that protein kinase C (PKC) signaling can trigger hallmark events of cell cycle withdrawal in intestinal epithelial cells, including downregulation of cyclin D1, induction of p21(Waf1/Cip1), and activation of the growth suppressor function of pocket proteins. In the current study, we compared the cell cycle- and PKC-specific effects of the vanilloid resiniferatoxin (RTX), its parent diterpene resiniferonol 9,13,14-ortho phenylacetate (ROPA), and the PKC agonist PMA in the IEC-18 non-transformed intestinal crypt cell line. ROPA and PMA were found to produce strikingly similar alterations in cell cycle progression and PKC activity in IEC-18 cells, although PMA was approximately 1000-fold more potent in producing these effects. Both agents induced a transient PKC-dependent blockade in G1---> S progression associated with transient downregulation of cyclin D1 and induction of p21(Waf1/Cip1). In contrast, RTX produced a prolonged PKC-independent cell cycle arrest in G(0)/G(1) phase which was maintained for longer than 24h. This arrest was vanilloid receptor-independent and associated with prolonged downregulation of cyclin D1 mRNA and protein, with little effect on levels of p21(Waf1/Cip1). Combined exposure to RTX and ROPA produced a sustained and complete cell cycle blockade in IEC-18 cells, associated with depletion of cyclin D1 and sustained enhancement of p21(Waf1/Cip1) levels. PMA, ROPA, RTX and the RTX/ROPA combination were capable of activating ERK1/2 signaling in IEC-18 cells, albeit with different kinetics. In contrast, only PMA and ROPA activated JNK1/2 and p38 in this system. Notably, some preparations of commercially obtained RTX produced effects indistinguishable from those of the RTX/ROPA combination, suggesting that certain batches of the compound may contain significant amounts of ROPA (or another PKC agonist activity). Together, these data demonstrate that structurally related compounds can produce similar cell cycle-specific effects but through distinct mechanisms. In addition, they add to a growing body of evidence that vanilloids can have antiproliferative effects in a variety of cell types. PMID- 15130765 TI - Synthesis and electrophysiological characterization of cyclic morphiceptin analogues. AB - A challenge in opioid peptide chemistry and pharmacology is the possibility to develop novel peptides with peripheral selectivity. An enzymatically stable opioid peptide could involve an antidiarrheal effect. For this reason, we constrained the highly selective and potent tetrapeptide morphiceptin with a 6 atom bridge, resulting in a cyclic amide and an ester analogue, 2 and 3, respectively. Taking advantage of the functional coupling of the opioid receptor with the heteromultimeric G-protein-coupled inwardly rectifying K+ (GIRK1/GIRK2) channel, either the wild-type mu-, kappa-, delta- or a mutated mu-opioid receptor (hMORS329A) was functionally co-expressed with GIRK1/GIRK2 channels and a regulator of G-protein signaling (RGS4) in Xenopus laevis oocytes. The two microelectrode voltage clamp technique was used to measure the opioid receptor activated GIRK1/GIRK2 channel responses. Both cyclic analogues were equally potent via the wild-type mu-opioid receptor hMORwt (EC(50) value 976.5 +/- 41.7 for 2 and 1017.7 +/- 60.7 for 3), while the EC(50) value for Tyr-Pro-Phe-D-Pro NH(2) measured 59.3 +/- 4.8 nM. These three agonists displayed a four to five times decreased potency via hMORS329A as compared to the wild type. Interestingly, no effect on kappa- and delta-opioid receptors was observed. The intramolecular bridge created by cyclization of morphiceptin prevents dipeptidyl peptidase IV from interacting with these analogues. We conclude that constraining morphiceptin with a 6-atom bridge resulted in enzymatically stable peptidomimetics that are exclusively active on mu-opioid receptors. These analogues provide an interesting template in the promising approach for the design of potential antidiarrheal agents. PMID- 15130766 TI - A cell-based system to identify and characterize the molecular mechanism of drug metabolizing enzyme (DME) modulators. AB - Many naturally occurred or synthetic compounds can modulate the body's drug metabolizing enzymes to enhance carcinogen detoxification, and some have demonstrated remarkable cancer prevention effects. Understanding the molecular mechanism behind each candidate agent is critically important in designing rational cancer chemoprevention strategies. In this work, we have employed a set of molecular mechanism-based assays and characterized eight classes of known drug metabolizing enzyme (DME) modulators in a cellular system. Examination of mRNA and protein levels of representative phase I and phase II enzymes validated the results obtained in our cell-based system. Our data confirmed that the antioxidant ethoxyquin (EQ) and the isothiolcyanate sulfurophane (SFP) exclusively activate the antioxidant response element (ARE), and thus represent monofunctional inducers. We were also able to reclassify some compounds, and to use the system to identify structure-activity relationships among structurally related but different compounds. Finally, this cell-based system permitted us to identify a potential novel mechanism for cross-talk between the ARE and the xenobiotic response element (XRE)-mediated pathways. PMID- 15130767 TI - Anti-proliferation effect of 3-amino-2-imino-3,4-dihydro-2H-1,3-benzothiazin-4 one (BJ-601) on human vascular endothelial cells: G0/G1 p21-associated cell cycle arrest. AB - The aim of this study was to examine the anti-proliferation effect of 3-amino-2 imino-3,4-dihydro-2H-1,3-benzothiazin-4-one (BJ-601) on human vascular endothelial cells and its possible molecular mechanism underlying. Our data showed that BJ-601 at a range of concentrations (0-40 microM) dose- and time dependently decreased cell number in cultured human dermal microvascular endothelial cells (HDMVECs), but not human fibroblasts. The BJ-601-induced growth inhibition in HDMVECs was reversible. [3H]thymidine incorporation demonstrated that BJ-601 arrested the HDMVECs at the G0/G1 phase of the cell cycle. Western blot analysis revealed that BJ-601 (0-40 microM) dose-dependently increased the levels of the protein p21, but not of p27, p53, cyclins A, D1, D3 and E, cyclin dependent kinase 2 (CDK2), and CDK4 in HDMVECs. Immunoprecipitation showed that the formation of the CDK2-p21 complex, but not CDK2-p27, CDK4-p21 and CDK4-p27 complexes, was increased in the BJ-601-treated HDMVECs. Kinase assay further demonstrated that CDK2, but not CDK4, kinase activity was decreased in a dose dependent manner in the BJ-601-treated HDMVECs. Pretreatment of HDMVECs with a p21 antisense oligonucleotide, which blocked the expression of p21 protein, reversed the BJ-601-induced inhibition of [3H]thymidine incorporation into HDMVECs. Moreover, cotreatment of the endothelial cells with protein kinase C (PKC) inhibitor, staurosporine, prevented the BJ-601-induced decrease of [3H]thymidine incorporation into HDMVECs. Administration of BJ-601 dose dependently inhibited capillary-like tube formation of HDMVECs in Matrigel. In conclusion, these data suggest that BJ-601 inhibits HDMVECs proliferation by increasing the level of p21 protein, which in turn inhibits CDK2 kinase activity, and finally causes retardation of the cell cycle at the G0/G1 phase. PMID- 15130768 TI - Cloning and characterization of mouse nucleoside triphosphate diphosphohydrolase 3. AB - We have cloned and characterized the nucleoside triphosphate diphosphohydrolase-3 (NTPDase3) from mouse spleen. Analysis of cDNA shows an open reading frame of 1587 base pairs encoding a protein of 529 amino acids with a predicted molecular mass of 58953Da and an estimated isoelectric point of 5.78. The translated amino acid sequence shows the presence of two transmembrane domains, eight potential N glycosylation sites and the five apyrase conserved regions. The genomic sequence is located on chromosome 9F4 and is comprised of 11 exons. Intact COS-7 cells transfected with an expression vector containing the coding sequence for mouse NTPDase3 hydrolyzed P2 receptor agonists (ATP, UTP, ADP and UDP) but not AMP. NTPDase3 required divalent cations (Ca2+ > Mg2+) for enzymatic activity. Interestingly, the enzyme had two optimum pHs for ATPase activity (pH 5.0 and 7.4) and one for ADPase activity (pH 8.0). Consequently, the ATP/ADP and UTP/UDP hydrolysis ratios were two to four folds higher at pH 5.0 than at pH 7.4, for both, intact cells and protein extracts. At pH 7.4 mouse NTPDase3 hydrolyzed ATP, UTP, ADP and UDP according to Michaelis-Menten kinetics with apparent K(m)s of 11, 10, 19 and 27 microM, respectively. In agreement with the K(m) values, the pattern of triphosphonucleoside hydrolysis showed a transient accumulation of the corresponding diphosphonucleoside and similar affinity for uracil and adenine nucleotides. NTPDase3 hydrolyzes nucleotides in a distinct manner than other plasma membrane bound NTPDases that may be relevant for the fine tuning of the concentration of P2 receptor agonists. PMID- 15130769 TI - Peripheral-type benzodiazepine receptor (PBR) and PBR drug ligands in fibroblast and fibrosarcoma cell proliferation: role of ERK, c-Jun and ligand-activated PBR independent pathways. AB - Peripheral-type benzodiazepine receptor (PBR) is a 18-kDa high-affinity drug and cholesterol binding protein, that has been implicated in several physiological processes, such as cholesterol transport and mitochondrial respiration. Specific PBR ligands regulate cell proliferation, although their action is controversial and probably cell-type specific. The aim of the present study was to examine the expression of PBR in cells of mesenchymal origin, i.e. human fibroblasts and fibrosarcoma cells, as well as its role in the regulation of their proliferation. Both mesenchymal cell types express high levels of PBR, localized exclusively in mitochondria. PBR-specific drug ligands, the isoquinoline carboxamide PK 11195 and the benzodiazepine Ro5-4864, at relative high concentrations (10(-4)M), exert a strong inhibitory effect on cell proliferation by arresting the cells at the G0/G1 phase of the cell cycle, while no apoptotic cell death was observed. In normal fibroblasts, this inhibition was correlated with a decrease in the activation of the cell cycle markers ERK and c-Jun. PBR knockdown by RNA inhibition did not affect the proliferation of either cell type and did not influence the inhibitory effect of PK 11195 and Ro5-4864 on cell growth. These data suggest that in fibroblasts and fibrosarcoma cells PBR drug ligands inhibit cell proliferation in a PBR-independent manner. These results are in contrast to data reported on cells of epithelial origin, suggesting that the origin of the cells is crucial in defining the role of PBR in their proliferation, and raise caution in the commonly made assumption that PBR mediates cell functions affected by PBR drug ligands. PMID- 15130770 TI - Effect of stereo and regiochemistry towards wild and multidrug resistant HIV-1 virus: viral potency of chiral PETT derivatives. AB - Chiral derivatives of several substituted halopyridyl and thiazolyl PETT compounds were synthesized as non-nucleoside inhibitors of the reverse transcriptase (RT) enzyme of the human immunodeficiency virus (HIV-1). Molecular modeling studies indicated that because of the asymmetric geometry of the non nucleoside inhibitors (NNRTI) binding pocket, the "R" stereoisomers would fit the NNRTI binding pocket of the HIV-1 RT much better than the corresponding "S" stereoisomers, as reflected by their 10(4)-fold lower K(i) values. The "R" stereoisomers of several PETT derivatives inhibited the recombinant RT in vitro with lower IC(50) values than their enantiomers. The active compounds were further evaluated for their ability to inhibit HIV-1 replication in human peripheral blood mononuclear cells (PBMCs). All the "R" isomers again showed potent anti-HIV activity and inhibited the replication of the HIV-1 strains HTLV(IIIB) in PBMCs at nanomolar concentrations whereas their enantiomers were less potent. The lead compounds for the respective groups were further tested against A17 (NNRTI-resistant, Y181C mutant RT), and A17Var (NNI-resistant Y181C +/- K103N mutant RT) as well as multidrug resistant viral strains. The results indicated that the lead compounds were several logs more potent than the standard NNRTI drug nevirapine. Structure-activity relationship among the derivatives showed preference of pyridyl unit with halo substitutions primarily at 5-position demonstrating the importance of both the stereochemistry as well as regiochemistry. Our data provides experimental evidence that the stereochemistry and the regiochemistry of non-nucleoside inhibitors can profoundly affect their anti-HIV activity. PMID- 15130771 TI - Roscovitine, olomoucine, purvalanol: inducers of apoptosis in maturing cerebellar granule neurons. AB - Cyclin-dependent kinases (CDKs) mediate proliferation and neuronal development, while aberrant CDK activity is associated with cancer and neurodegeneration. Consequently, pharmacologic inhibitors, such as 2,6,9-trisubstituted purines, which potently inhibit CDKs 1, 2, and 5, were developed to combat these pathologies. One agent, R-roscovitine (CYC202), has advanced to clinical trials as a potential cancer therapy. In primary neuronal cultures, these agents have been used to delineate the physiologic and pathologic functions of CDKs, and associated signaling pathways. Herein we demonstrate that three 2,6,9 trisubstituted purines: olomoucine, roscovitine, and purvalanol, used at concentrations ascribed by others to potently inhibit CDKs 1, 2, and 5, are powerful triggers of death in maturing cerebellar granule neurons, assessed by loss of mitochondrial reductive capacity and differential staining with fluorescent indicators of living/dead neurons. Based on several criteria, including delayed time course and establishment of an irreversible commitment point of death, pyknotic cell and nuclear morphology, and caspase-3 cleavage, the death process is apoptotic. However, pharmacological and biochemical data indicate that apoptosis is independent of CDK 1, 2, or 5 inhibition. This is based on the pattern of changes in c-jun mRNA, c-Jun protein, and Ca(2+)/cAMP response element binding protein (CREB) phosphorylation, and also, the ineffectiveness of structurally distinct CDK 1, 2, and 5 inhibitors butyrolactone 1 and PNU112445A to induce apoptosis. Collectively, our results, and those of others, indicate that the CDK regulation of transcription (CDKs 7 and 9) should be examined as a target of these agents, and as an indirect mediator of neuronal fate. PMID- 15130772 TI - Dopamine toxicity involves mitochondrial complex I inhibition: implications to dopamine-related neuropsychiatric disorders. AB - Dopamine, which is suggested as a prominent etiological factor in several neuropsychiatric disorders such as Parkinson's disease and schizophrenia, demonstrates neurotoxic properties. In such dopamine-related diseases mitochondrial dysfunction has been reported. Dopamine oxidized metabolites were shown to inhibit the mitochondrial respiratory system both in vivo and in vitro. In the present study, we suggest an additional mechanism for dopamine toxicity, which involves mitochondrial complex I inhibition by dopamine. In human neuroblastoma SH-SY5Y cells dopamine induced a reduction in ATP concentrations, which was negatively correlated to intracellular dopamine levels (r = - 0.96, P = 0.012), and was already evident at non-toxic dopamine doses. In disrupted mitochondria dopamine inhibited complex I activity with IC50 = 11.87 +/- 1.45 microm or 8.12 +/- 0.75 microM in the presence of CoQ or ferricyanide, respectively, with no effect on complexes IV and V activities. The catechol moiety, but not the amine group, of dopamine is essential for complex I inhibition, as is indicated by comparing the inhibitory potential of functionally and structurally dopamine-related compounds. In line with the latter is the finding that chelatable FeCl2 prevented dopamine-induced inhibition of complex I. Monoamine oxidase A and B inhibitors, as well as the antioxidant butylated hydroxytoluene (BHT), did not prevent dopamine-induced inhibition, suggesting that dopamine oxidation was not involved in this process. The present study suggests that dopamine toxicity involves, or is initiated by, its interaction with the mitochondrial oxidative phosphorylation system. We further hypothesize that this interaction between dopamine and mitochondria is associated with mitochondrial dysfunction observed in dopamine-related neuropsychiatric disorders, such as schizophrenia and Parkinson's disease. PMID- 15130773 TI - Activation of nuclear factor kappa B by diesel exhaust particles in mouse epidermal cells through phosphatidylinositol 3-kinase/Akt signaling pathway. AB - Diesel exhaust particles (DEP) induce intense inflammatory and allergic immune responses. The epidermal cells receive much exposure to DEP, and are an important source of pro-inflammatory cytokines and other inflammatory mediators. Transcription factors, such as nuclear factor kappa B (NF-kappaB) and activator protein 1 (AP-1), regulate the expression of these mediators. We hypothesize that the transcription factors are target of DEP action. The current study sought to determine whether DEP-activated NF-kappaB and AP-1 in a mouse epidermal cell line, JB6 P(+) cells. Using stable transfectants of JB6 P(+) cells expressing NF kappaB or AP-1 luciferase reporter constructs, we demonstrated that exposure to DEP at a non-cytotoxic concentration significantly enhanced the transactivation of NF-kappaB, but not AP-1. Furthermore, DEP promoted phosphorylation of Akt, a substrate of phosphatidylinositol 3-kinase (PI3K), on Ser-473 and Thr-308 in a PI3K-dependent manner, and enhanced phosphorylation of down-stream p70/p85 S6 kinases (p70/p85S6K) as well as glycogen synthase kinase-3beta (GSK-3beta). Blockage of PI3K activation eliminated DEP-stimulated NF-kappaB transactivation. Although SAPK/JNK pathway was modestly activated by DEP, it was not involved in NF-kappaB transactivation. DEP had little effect on the phosphorylation of ERKs and p38 MAPK. Thus, DEP-induced transactivation of NF-kappaB is mediated by PI3K/Akt signaling pathway. PMID- 15130774 TI - 4-Hydroxy tempol-induced impairment of mitochondrial function and augmentation of glucose transport in vascular endothelial and smooth muscle cells. AB - The water-soluble and cell permeable nitroxide derivative 4-hydroxy tempol (TPL) has been shown to reduce or ameliorate oxidative stress-induced dysfunction and damage in vascular endothelial cells. We studied the effects of TPL on glucose transport and metabolism in bovine aortic endothelial (VEC) and smooth muscle cells (VSMC) under normal and high glucose conditions. Normally, these cells operate an autoregulatory protective mechanism that limits the rate of glucose transport under hyperglycemic conditions by decreasing the cell content of their typical glucose transporter GLUT-1 mRNA and protein as well as its plasma membrane abundance. TPL augmented the rate of glucose transport both under normo- and hyperglycemic conditions by increasing GLUT-1 mRNA and protein content and its plasma membrane abundance in both types of cells, leading to an increased flux of glucose into the cells. These effects were found related to ROS generating and oxidant activities of TPL and to a decreased rate of mitochondrial ATP production under both normo- and hyperglycemic conditions. Since impaired mitochondrial functions, and in particular decreased rate of ATP production, augment the expression of GLUT-1 protein and glucose transport and metabolism, we suggest that the stimulatory effects of TPL in vascular cells results from its unfavorable interactions in the mitochondrion. It is therefore suggested that effects of TPL in cells of cardiovascular system be evaluated in parallel to its adverse effects on glucose and energy metabolism. PMID- 15130775 TI - Rosiglitazone increases extravasation of macromolecules and endothelial nitric oxide synthase in skeletal muscles of the fructose-fed rat model. AB - Reduced extravasation of macromolecules in skeletal muscle has recently been documented in the fructose-fed rat model, corroborating a hypothesis that a functional obliteration of muscle regional microcirculation might lead to hypertension and restrict access of nutrients and hormones to their target cells. The goal of this study was to assess the impact of a treatment with rosiglitazone on the reduced muscle vasopermeability observed previously in the fructose-fed rat model. Fructose-fed Sprague-Dawley rats were gavaged with rosiglitazone (10 micromol kg(-1) per day; n = 21) or the vehicle only (n = 19) for 3 consecutive weeks before assessing the extravasation of Evans Blue (EB) dye in vivo in distinct muscle groups. Relative to control group, rosiglitazone reduced mean arterial blood pressure (Delta = -16.7%, P < 0.001), plasma insulin (Delta= 39.1%, P < 0.05) and plasma triglyceride (Delta= -32.8 %, P < 0.01) concentrations in a significant manner. Plasma VEGF concentrations were significantly lower in the rosiglitazone-treated animals compared to the control animals (32.7 +/- 0.8 pg ml(-1) versus 46.1 +/- 1.2 pg ml(-1), P < 0.001). While no changes were observed in the lungs or the kidneys, fructose-fed rats treated with rosiglitazone had a 30-50% increase (P < 0.005) in the extravasation of EB regardless of the skeletal muscle group studied (rectus femoris, soleus, gastrocnemius lateralis, vastus lateralis and tibialis cranalis). In homogenates of skeletal muscles (vastus lateralis) of fructose-fed rats, rosiglitazone resulted in a significant increase in NO synthase (NOS) activity (Delta = +41.9 %, P < 0.003) as well as endothelial NOS immunoreactive mass (Delta = +37.8 %, P < 0.01) compared to the control animals. There was no change in the immunoreactive level of the nNOS isoform, the most abundant muscle isoform, or in the immunoreactive levels of VEGF. In conclusion, rosiglitazone appears to restore a vascular dysfunction previously documented in the skeletal muscle microcirculation, as evidenced by improved skeletal muscle vasopermeability and upregulation of the muscle endothelium-NO system in the fructose-fed rat model. These effects on muscle per se might also result in a partial improvement of the insulin resistance phenomenon by improving the distribution of nutrients and insulin to skeletal muscle. This effect appears to be independent of circulating levels of VEGF since changes in plasma concentrations of this permeability factor were lower in the rosiglitazone-treated group. PMID- 15130776 TI - Adenosine induces apoptosis in the human gastric cancer cells via an intrinsic pathway relevant to activation of AMP-activated protein kinase. AB - Extracellular adenosine significantly reduced cell viability in a dose (0.1-20mM) and treatment time (24-72h)-dependent manner in GT3-TKB cells, a human gastric cancer cell line. Nuclei of cells were reactive to Hoechst 33342, a marker of apoptosis, and an anti-single-stranded DNA. Adenosine-induced GT3-TKB cell death was significantly inhibited by dipyridamole, an inhibitor of adenosine transporter, and 5'-amino-5'-deoxyadenosine, an inhibitor of adenosine kinase, but the effect was not affected by theophylline, a broad inhibitor of adenosine receptors, 8-cyclopentyltheophylline, an inhibitor of A(1) adenosine receptors or 3,7-dimethyl-1-propargylxanthine, an inhibitor of A(2a) adenosine receptors. Adenosine had no effect on mitochondrial membrane potentials. The effect of adenosine on GT3-TKB cell death was not inhibited by a pancaspase inhibitor or inhibitors of caspase-1,-3,-4,-8, and -9. 5-Aminoimidazole-4-carboxamide ribonucleoside (AICAR), an activator of AMP-activated protein kinase (AMPK), significantly reduced GT3-TKB cell viability, but the AICAR action was not reinforced in the presence of adenosine. The results of the present study, thus, suggest that extracellular adenosine induces apoptosis in GT3-TKB cells by its uptake into cells and conversion to AMP followed by activation of AMPK, regardless of caspase activation linked to the mitochondria and the endoplasmic reticulum. PMID- 15130777 TI - Molten-globule like partially folded states of human serum albumin induced by fluoro and alkyl alcohols at low pH. AB - Human serum albumin (HSA) exists in a molten-globule like state at low pH (pH 2.0). We studied the effects of trifluoroethanol (TFE) and hexafluoroisopropanol (HFIP) on the acid-denatured state of HSA by far-UV circular dichroism (CD), near UV CD, tryptophan fluorescence, and 1-anilinonaphthalene-8-sulfonic acid (ANS) binding. At pH 2.0, these alcohols induced the formation of alpha-helical structure as evident from the increase in mean residue ellipticity (MRE) value at 222 nm. On addition of different alcohols, HSA exhibited a transition from the acid-denatured state to the alpha-helical state and loss of ANS-binding sites reflected by the decrease in ANS fluorescence at 480 nm. However, the concentration range required to bring about the transition varied greatly among different alcohols. HFIP was found to have highest potential whereas methanol was least effective in inducing the transition. The order of effectiveness of alcohols was shown to be: HFIP > TFE > 2-chloroethanol > tert-butanol > isopropanol > ethanol > methanol as evident from the Cm values. The near-UV CD spectra and tryptophan fluorescence showed the differential effects of halogenated alcohols with those of alkanols. A comparison of the m values, showing the dependence of Delta GH on alcohol concentration, suggests that the helix stabilizing potential of different alcohols is due to the additive effect of different constituent groups present whereas remarkably higher potential of HFIP involves some other factor in addition to the contribution of constituent groups. PMID- 15130778 TI - Xanthine oxidase activates pro-matrix metalloproteinase-2 in cultured rat vascular smooth muscle cells through non-free radical mechanisms. AB - Reactive oxygen species (ROS) have been implicated in the regulation of matrix metalloproteinases (MMPs). The xanthine/xanthine oxidase (X/XO) reaction has been widely used as a source of exogenous ROS in studying MMPs, but commercial XO has also been known to be contaminated by proteolytic activity, and MMPs are protease sensitive substrate. We have investigated the activation of proMMP-2 by X/XO in cultured vascular smooth muscle cells (SMCs). SMCs were incubated with X/XO (unpurified or purified) or XO alone for 24h. X/XO activated proMMP-2 in a dose dependent manner. A similar profile was observed using XO. Purified XO produced lower amounts of active MMP-2 compared to unpurified XO. EPR study showed that X/XO, not XO itself, produced superoxide anion, which was completely scavenged by SOD. However, X/XO-induced proMMP-2 activation could not be inhibited by combination of SOD and catalase. Incubation with XO either in cell-free conditioned media or in cells resulted in similar amounts of active MMP-2, suggesting that membrane-type-MMPs were not involved in proMMP-2 activation. This was further confirmed by the lack of inhibitory effect of hydroxamate MMP inhibitor, BB1101. Aprotinin blocked unpurified XO-induced proMMP-2 activation in a dose-dependent manner, demonstrating the proteolytic activity contained in XO is essential. We conclude that proteolytic activity contained in XO, rather the ROS derived from X/XO, is responsible for proMMP-2 activation in cultured SMCs. The results also suggest that caution needs to be taken when interpreting the reported results on activation of MMPs where X/XO had been used as an "authentic" source of superoxide anion. PMID- 15130779 TI - Hyperexpression of N-acetylglucosaminyltransferase-III in liver tissues of transgenic mice causes fatty body and obesity through severe accumulation of Apo A-I and Apo B. AB - N-Acetylglucosaminyltransferase (GnT)-III catalyzes the attachment of an N acetylglucosamine (GlcNAc) residue to mannose in beta(1-4) configuration in the region of N-glycans and forms a bisecting GlcNAc. To investigate the pathophysiological role of dysregulated glycosylation mediated by aberrantly expressed GnT-III, we generated transgenic mice hyperexpressing the human GnT-III in the liver by introducing human GnT-III cDNA under the control of mouse albumin enhancer/promoter. Total five transgenic founder mice (pGnTSVTpA-10, -14, -20, 25, and -51) expressed the human GnT-III in their livers and were characterized by molecular genetic means. The copy number of transgene integrated into the genome of these mice ranged between 1 and 3 copies per haploid genome. Northern and Western blot analyses showed that the transgene is specifically expressed in the liver but not in any other tissues tested. The triglyceride level in GnT-III transgenic mice was significantly decreased, however, no significant differences in the levels of glucose, cholesterol, or albumin were observed between transgenic and nontransgenic mice. Although glutamate oxaloacetic transaminase and glutamic pyruvic transaminase activities of transgenic mice were also higher than those of nontransgenic mice, no differences in total bililubin and total protein were observed between the two animal lines. Large amounts of apolipoprotein (Apo) A-I and Apo B were specifically detected in the intracellular liver of transgenic mice. The accumulation of Apo A-I in hepatocytes may be due to aberrant glycosylation, since glycosylated Apo A-I was not observed in transgenic mice. However, the accumulated Apo B was severely glycosylated. Therefore, it is suggested that highly expressed transgenic GnT-III allowed unknown target proteins to be glycosylated in large amounts, and the resulting target protein(s) disrupted in assembly formation of Apo A-I in the hepatocytes and cause a decrease in the release of lipoproteins and accumulations of Apo A-I and Apo B in the liver. The transgenic mice showed aberrant glycosylation by GnT-III, resulting in numerous lipid droplets in liver tissues and the obesity. These mice showed microvesicular fatty changes with abnormal lipid accumulation in the hepatocytes. Our study provides the basis for future analysis of the role of glycosylation in hepatic pathogenesis. In the transgenic mice, Apo A-I and Apo B were significantly increased compared with levels in nontransgenic liver tissues. PMID- 15130780 TI - Oligomerization is required for betaine-homocysteine S-methyltransferase function. AB - Betaine-homocysteine methyltransferase (BHMT) is a member of a family (Pfam 02574) of zinc- and thiol/selenol-dependent methyltransferases. All family members purified to date are monomers, except BHMT, which is an oligomer. We have studied how C-terminal truncation or mutagenic replacement of residues within or associated with the unique dimerization arm of this enzyme affects oligomerization and function. Two C-terminal truncation mutants, S325 and D371, do not express well in Escherichia coli and are inactive. Residues within the dimerization arm (H338, R346, W352, R361, P362, Y363, N364, and P365) and one that forms a hydrogen bond to the arm (E266) were changed to alanine. All mutants maintained a normal or near-normal ability to bind zinc. E266A, R361A, P362A, Y363A, N364A, and P365A displayed near-normal catalytic activity, but H338A had only 10% of the wild-type enzyme activity. Like the wild-type enzyme, most mutants eluted as tetramers from gel filtration columns and formed discrete bands on SDS-PAGE gels following glutaraldehyde crosslinking. Mutants R346A and W352A had negligible activity, eluted as dimers, and displayed aberrant crosslinking properties. These data indicate that unlike other Pfam 02574 members, oligomerization of BHMT is required for function. PMID- 15130781 TI - The role of cysteine 160 in thiamine diphosphate binding of the Calvin-Benson Bassham cycle transketolase of Rhodobacter sphaeroides. AB - The transketolase gene (cbbT) that encodes the Calvin-Benson-Bassham pathway transketolase (CbbT) of Rhodobacter sphaeroides was overexpressed in Escherichia coli and the recombinant protein purified to homogeneity. Like other transketolases, R. sphaeroides CbbT was found to be inactivated in the presence of oxygen. At its optimal pH of 7.8, CbbT displays a specific activity of 37 U/mg, a KR5P of 949 microM, a KXu5P of 11 microM, and a KThDP of 1.8 microM. Cysteine 160, equivalent to Cys159 of the yeast enzyme, is found within the active site and is loosely conserved amongst several sources of transketolase. To investigate the role of cysteine 160 found in the active site of R. sphaeroides CbbT, this residue was targeted for mutagenesis. Cys160 was changed to alanine, serine, aspartate, and glutamate. To compare the effect of these mutations on ThDP binding, spectral techniques were employed in addition to analysis by enzymatic activity. Fluorescence quenching was used to measure both equilibrium binding constants as well as first order rates of binding. The results of these studies indicated that Cys160 played an important and substantial role in cofactor binding, revealing the importance of this loosely conserved residue. In addition, the Cys160 mutants did not appear to alter oxygen-mediated inactivation. PMID- 15130782 TI - Identification of UGT2B9*2 and UGT2B33 isolated from female rhesus monkey liver. AB - Two UDP-glucuronosyltransferases (UGT2B9(*)2 and UGT2B33) have been isolated from female rhesus monkey liver. Microsomal preparations of the cell lines expressing the UGTs catalyzed the glucuronidation of the general substrate 7-hydroxy-4 (trifluoromethyl)coumarin in addition to selected estrogens (beta-estradiol and estriol) and opioids (morphine, naloxone, and naltrexone). UGT2B9(*)2 displayed highest efficiency for beta-estradiol-17-glucuronide production and did not catalyze the glucuronidation of naltrexone. UGT2B33 displayed highest efficiency for estriol and did not catalyze the glucuronidation of beta-estradiol. UGT2B9(*)2 was found also to catalyze the glucuronidation of 4-hydroxyestrone, 16 epiestriol, and hyodeoxycholic acid, while UGT2B33 was capable of conjugating 4 hydroxyestrone, androsterone, diclofenac, and hyodeoxycholic acid. Three glucocorticoids (cortisone, cortisol, and corticosterone) were not substrates for glucuronidation by liver or kidney microsomes or any expressed UGTs. Our current data suggest the use of beta-estradiol-3-glucuronidation, beta-estradiol-17 glucuronidation, and estriol-17-glucuronidation to assay UGT1A01, UGT2B9(*)2, and UGT2B33 activity in rhesus liver microsomes, respectively. PMID- 15130783 TI - Role of hepatocyte nuclear factor 3 gamma in the expression of human CYP2C genes. AB - Hepatocyte nuclear factor 3 gamma (HNF-3 gamma) is an important transcription factor for the maintenance of specific liver functions. However, its relevance in the expression of human cytochrome P450 (CYP) genes has not yet been explored. Several HNF3 putative binding sites can be identified in human CYP2C 5'-flanking regions. Gene reporter experiments with proximal promoters revealed that HNF-3 gamma transactivated CYP2C8, CYP2C9, and CYP2C19 (25-, 4-, and 4-fold, respectively), but it did not transactivate CYP2C18. However, overexpression of HNF-3 gamma in hepatoma cells by means of a recombinant adenovirus induced CYP2C9, CYP2C18, and CYP2C19 mRNA (4.5-, 20-, and 50-fold, respectively) but did not activate endogenous CYP2C8. The lack of effect of HNF-3 gamma on endogenous CYP2C8 could be reversed by treating cells with the deacetylase inhibitor, trichostatin A, suggesting the existence of chromatin condensation around functional HNF3 elements in this gene. We conclude that HNF3 gamma is an important transcription factor for the hepatic-specific expression of human CYP2C genes. Our results also evidence that efficient transfection tools, such as adenoviral vectors, may be decisive for assessing the role of transcription factor on chromatin organized genes. PMID- 15130784 TI - Leptin deficiency prevents the activation of the murine alpaha 2(I) collagen promoter by acetaldehyde. AB - Leptin enhances hepatic fibrosis induced experimentally by various agents. The influence of leptin deficiency on murine alpha 2(I) collagen promoter activation by acetaldehyde and by TGF beta 1 was investigated. Acetaldehyde failed to activate the promoter in ob/ob stellate cells as compared to its activating effect in wild-type cells. By contrast, TGF beta 1 increased the activity of the promoter in the ob/ob and wild-type cells. Total cell TGF beta 1 and secretion of total and free TGF beta 1 were lower in cultured stellate cells from ob/ob mice than wild-type mice. Acetaldehyde increased free cell TGF beta1 and secretion of total and free TGF beta 1 in wild-type, but not in ob/ob cells. The lack of activation of the alpha 2(I) collagen promoter by acetaldehyde in association with a lack of increase in TGF beta 1 in response to acetaldehyde in ob/ob stellate cells provides further evidence for a mediating role of TGF beta 1 in the actions of acetaldehyde on collagen transcription. PMID- 15130785 TI - Maximization of the rate of chloride conduction in the CFTR channel pore by ion ion interactions. AB - Multi-ion pore behaviour has been identified in many Cl(-) channel types but its biophysical significance is uncertain. Here, we show that mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel that disrupt anion-anion interactions within the pore are associated with drastically reduced single channel conductance. These results are consistent with models suggesting that rapid Cl(-) permeation in CFTR results from repulsive ion-ion interactions between Cl(-) ions bound concurrently inside the pore. Naturally occurring mutations that disrupt these interactions can result in cystic fibrosis. PMID- 15130786 TI - Effects of selenium on altered mechanical and electrical cardiac activities of diabetic rat. AB - Since selenium compounds can restore some metabolic parameters and structural alterations of diabetic rat heart, we were tempted to investigate whether these beneficial effects extend to the diabetic rat cardiac dysfunctions. Diabetes was induced by streptozotocin (50mg/kg body weight) and rats were then treated with sodium selenite (5 micromol/kg body weight/day) for four weeks. Electrically stimulated isometric contraction and intracellular action potential in isolated papillary muscle strips and transient (I(to)) and steady state (I(ss)) outward K(+) currents in isolated cardiomyocytes were recorded. Sodium selenite treatment could reverse the prolongation in both action potential duration and twitch duration of the diabetic rats, and also cause significant increases in the diminished amplitudes of the two K(+) currents. Treatment of rats with sodium selenite also markedly increased the depressed acid-soluble sulfhydryl levels of the hearts. Our data suggest that the beneficial effects of sodium selenite treatment on the mechanical and electrical activities of the diabetic rat heart appear to be due to the restoration of the diminished K(+) currents, partially, related to the restoration of the cell glutathione redox cycle. PMID- 15130787 TI - Kinetics of oxygen binding to ferrous myeloperoxidase. AB - Myeloperoxidase (MPO), which is involved in host defence and inflammation, is a unique peroxidase in having a globin-like standard reduction potential of the ferric/ferrous couple. Intravacuolar and exogenous MPO released from stimulated neutrophils has been shown to exist in the oxyferrous form, called compound III. To investigate the reactivity of ferrous MPO with molecular oxygen, a stopped flow kinetic analysis was performed. In the absence of dioxygen, ferrous MPO decays to ferric MPO (0.04 s(-1) at pH 8 versus 1.4 s(-1) at pH 5). At pH 7.0 and 25 degrees C, compound III formation (i.e., binding of dioxygen to ferrous MPO) occurs with a rate constant of (1.1+/-0.1) x 10(4)M(-1)s(-1). The rate doubles at pH 5.0 and oxygen binding is reversible. At pH 7.0, the dissociation equilibrium constant of the oxyferrous form is (173+/-12)microM. The rate constant of dioxygen dissociation from compound III is much higher than conversion of compound III to ferric MPO (which is not affected by the oxygen concentration). This allows an efficient transition of compound III to redox intermediates which actually participate in the peroxidase or halogenation cycle of MPO. PMID- 15130790 TI - Profound normalisation challenges remain in the analysis of data from microarray experiments. PMID- 15130791 TI - Overcoming confounded controls in the analysis of gene expression data from microarray experiments. AB - A potential limitation of data from microarray experiments exists when improper control samples are used. In cancer research, comparisons of tumour expression profiles to those from normal samples is challenging due to tissue heterogeneity (mixed cell populations). A specific example exists in a published colon cancer dataset, in which tissue heterogeneity was reported among the normal samples. In this paper, we show how to overcome or avoid the problem of using normal samples that do not derive from the same tissue of origin as the tumour. We advocate an exploratory unsupervised bootstrap analysis that can reveal unexpected and undesired, but strongly supported, clusters of samples that reflect tissue differences instead of tumour versus normal differences. All of the algorithms used in the analysis, including the maximum difference subset algorithm, unsupervised bootstrap analysis, pooled variance t-test for finding differentially expressed genes and the jackknife to reduce false positives, are incorporated into our online Gene Expression Data Analyzer ( http:// bioinformatics.upmc.edu/GE2/GEDA.html ). PMID- 15130792 TI - Application of z-score transformation to Affymetrix data. AB - Z-score transformation has been successfully used as a normalisation procedure for microarray data generated using radioactively labelled probes with spotted cDNA arrays. One of the advantages of the z-score transformation method is that it provides a way of standardising data across a wide range of experiments and allows the comparison of microarray data independent of the original hybridisation intensities. The feasibility of applying z-score transformation to other types of linear microarray data, specifically that generated using fluorescently labelled probes with Affymetrix chips, was tested in three separate scenarios and is discussed here. In the first scenario, Affymetrix data from the NCBI (National Center for Biotechnology Information) GEO (Gene Expression Omnibus) database was used to demonstrate that z-score transformation preserved the essential phylogenetic grouping between primate species' fibroblast gene expression baseline measurements. The second scenario employed z-score transformation on data consisting of a series of genes spiked-in at known concentrations and arrayed in a Latin square format. We were able to reconstruct the entire set of spike-in concentration curves without prior knowledge of their format by using z-score transformation as the normalisation process. Finally, we show that z-score transformed data maintains the integrity of separate samples from different experiments and laboratories, as demonstrated by accurate grouping of clustered data according to sample identity. We conclude that data normalised by z-score transformation can be easily used with Affymetrix data without noticeable loss of information content. Z-score transformation provides a useful tool for comparisons between experiments and between laboratories that use the Affymetrix platform. PMID- 15130793 TI - Clinically validated benchmarking of normalisation techniques for two-colour oligonucleotide spotted microarray slides. AB - Acquisition of microarray data is prone to systematic errors. A correction, called normalisation, must be applied to the data before further analysis is performed. With many normalisation techniques published and in use, the best way of executing this correction remains an open question. In this study, a variety of single-slide normalisation techniques, and different parameter settings for these techniques, were compared over many replicated microarray experiments. Different normalisation techniques were assessed through the distribution of the standard deviation of replicates from one biological sample across different slides. It is shown that local normalisation outperformed global normalisation, and intensity-based 'LOWESS' outperformed trimmed mean and median normalisation techniques. Overall, the top performing normalisation technique was a print-tip based LOWESS with zero robust iterations. Lastly, we validated this evaluation methodology by examining the ability to predict oestrogen receptor-positive and negative breast cancer samples with data that had been normalised using different techniques. PMID- 15130794 TI - The statistical distribution of the intensity of pixels within spots of DNA microarrays: what is the appropriate single-value representative? AB - This paper opens a discussion about an important issue in the analysis of data from spotted DNA microarrays: how to summarise into a single value the distribution for the intensity values of the pixels within a spot. Although the most popular statistic used is the median, there is no clear study demonstrating why it is more appropriate than other measures of central tendency such as the mean or the mode. Here, we argue that the median intensity is not the most appropriate measure for many common cases and discuss a frequently encountered case of a 'doughnut'-shaped spot for which the mode is closest to the 'expected' spot intensity. For an 'ideal' spot with a clear boundary and uniformly hybridised, the intensity of its pixels should approximately be normally distributed. In practical situations, these two requirements are often not met due to the physical properties of pins and the particularities of the printing and hybridisation processes. As a consequence, the distribution of the intensity of the pixels is usually negatively skewed. This asymmetry results in a larger displacement for the mean and median than for the mode from the ideal situation mentioned above. PMID- 15130795 TI - MicroPreP: a cDNA microarray data pre-processing framework. AB - The user-friendly MicroPreP framework was developed to transform raw intensity data from cDNA microarrays into high-quality data. The main features of this software are: LOWESS normalisation; merging of DNA microarray data from changing slide versions; outlier detection; and slide quality assessment. PMID- 15130796 TI - Automated DNA chip annotation tables at IFOM: the importance of synchronisation and cross-referencing of sequence databases. AB - The increasing popularity of DNA chip technology for the study of gene expression is producing, for each experiment, a sizable quantity of numerical data to analyse and an accompanying large number of gene identifiers that should be associated with the relevant biological annotation. We describe here a website at IFOM (FIRC Institute of Molecular Oncology) where we release regularly updated annotation tables for the most used Affymetrix oligonucleotide DNA chips and for the whole Research Genetics 46K clone collection for cDNA arrays. These tables are synchronised with every new release of the mouse and human UniGene databases (NCBI; National Center for Biotechnology Information), allowing fast and easy preliminary annotation of DNA array experiments. We also report some comparative evidence about the importance of biological database synchronisation and cross references in the process of generating annotation tables for DNA chips. PMID- 15130797 TI - Gene identification through large-scale EST sequence processing. AB - The technology of sequencing expressed sequence tags (ESTs) offers a relatively cheap alternative to whole genome sequencing and has become a valuable resource for gene discovery. The inherent characteristics of ESTs, such as transcript redundancy, low sequence quality and high error rates, require processing of the sequences before any gene prediction can be made. The process includes EST pre processing, analysis and similarity searches, and the data are generally stored in a database to organise the results and thereby assist the search for interesting genes. PMID- 15130798 TI - Are you my mother? Bayesian phylogenetic inference of recombination among putative parental strains. AB - Reconstructing evolutionary relationships using Bayesian inference has become increasingly popular due to the ability of Bayesian inference to handle complex models of evolution. In this review we concentrate on inference of recombination events between strains of viruses when these events are sporadic, ie rare relative to point mutations. Bayesian inference is especially attractive in the detection of recombination events because it allows for simultaneous inferences about the presence, number and location of crossover points and the identification of parental sequences. Current frequentist recombination identification falls into a sequential testing trap. The most likely parental sequences and crossover points are identified using the data and then the certainty of recombination is assessed conditional on this identification. After briefly outlining basic phylogenetic models, Bayesian inference and Markov chain Monte Carlo (MCMC) computation, we summarise three different approaches to recombination detection and discuss current challenges in applying Bayesian phylogenetic inference of recombination. PMID- 15130799 TI - FBSA: feature-based sequence alignment technique for very large sequences. AB - The ability to align pairs of very large molecular sequences is essential for a range of comparative genomic studies. However, given the complexity of genomic sequences, it has been difficult to devise a systematic method that can align - even within the same species - pairs of large sequences. Most existing approaches typically attempt to align nucleotide sequences while ignoring valuable features contained within them, eg they filter out low-complexity regions and retroelements before aligning the sequences. However, features are then added post-alignment for visualisation and analysis purposes. We argue that repetitive elements and other features (such as genes, exons and regulatory elements) should be part of the alignment process. A hierarchical approach that aligns the biologically relevant features before aligning the detailed nucleotide sequences has a number of interesting characteristics: (1) features define 'alignment anchor points' that can guide meaningful nucleotide alignment; (2) features can be weighted; (3) a hierarchical approach would identify only meaningful regions to be aligned; (4) nucleotide sequences can be described as sequences of features and non-features, providing a natural mechanism to divide the sequences for processing; and (5) computational speed is significantly faster than other approaches. In this paper, we describe and discuss a feature-based approach to aligning large genome sequences. We refer to this as 'feature-based sequence alignment'. PMID- 15130800 TI - SNPCEQer II: the integrated detection and analysis of SNPs in DNA sequences. AB - SNPCEQer II is a graphical user interface (GUI)-based application that integrates single nucleotide polymorphism (SNP) detection, SNP analysis and SNP editing in the Microsoft Windows (R) environment. SNPCEQer II detects SNPs in DNA sequences generated by the Beckman CEQ TM 2000 XL DNA analysis system. It provides tools to analyse SNPs by inspecting and comparing trace data (chromatograms) around putative SNPs with that of other related DNA sequences, and it can search for those SNPs in the National Center for Biotechnology Information (NCBI) databases. SNPCEQer II can determine the mutation type of a coding SNP and generate data for submission to the dbSNP database. The SNP report can be edited and printed, as can the chromatograms. SNPCEQer II is implemented in Visual C++. PMID- 15130801 TI - MAPPP: MHC class I antigenic peptide processing prediction. AB - MAPPP is a bioinformatics tool for the prediction of potential antigenic epitopes presented on the cell surface by major histocompatibility complex class I (MHC I) molecules to CD8 positive T lymphocytes. It combines existing predictions for proteasomal cleavage with peptide anchoring to MHC I molecules. PMID- 15130802 TI - Hetero: a program to simulate the evolution of DNA on a four-taxon tree. AB - We present a computer program to simulate the evolution of a nucleotide sequence on a phylogenetic tree with four tips. The program, Hetero, allows users to assign lineage-specific differences in the rate matrices used to describe the evolutionary process. It has a simple user interface and output, making it equally useful in the teaching and research of phylogenetics. PMID- 15130803 TI - MICAS: a fully automated web server for microsatellite extraction and analysis from prokaryote and viral genomic sequences. AB - MICAS is a web server for extracting microsatellite information from completely sequenced prokaryote and viral genomes, or user-submitted sequences. This server provides an integrated platform for MICdb (database of prokaryote and viral microsatellites), W-SSRF (simple sequence repeat finding program) and Autoprimer (primer design software). MICAS, through dynamic HTML page generation, helps in the systematic extraction of microsatellite information from selected genomes hosted on MICdb or from user-submitted sequences. Further, it assists in the design of primers with the help of Autoprimer, for sequences containing selected microsatellite tracts. PMID- 15130804 TI - AMID: autonomous modeler of intragenic duplication. AB - Intragenic duplication is an evolutionary process where segments of a gene become duplicated. While there has been much research into whole-gene or domain duplication, there have been very few studies of non-tandem intragenic duplication. The identification of intragenically replicated sequences may provide insight into the evolution of proteins, helping to link sequence data with structure and function. This paper describes a tool for autonomously modelling intragenic duplication. AMID provides: identification of modularly repetitive genes; an algorithm for identifying repeated modules; and a scoring system for evaluating the modules' similarity. An evaluation of the algorithms and use cases are presented. PMID- 15130805 TI - STORM towards protein function: systematic tailored ORF-data retrieval and management. AB - STORM (systematic tailored ORF-data retrieval and management) combines protein analyses of BLAST, Fasta, Pfam and ProtParam on a batch file of protein sequences. It subsequently summarises and organises the output in an Access database format. PMID- 15130806 TI - Genomic Object Net: I. A platform for modelling and simulating biopathways. AB - Genomic Object Net (GON) 1.0 is a software package for creating models and simulations of biopathways. Its core architecture employs the notion of a hybrid functional Petri net with extension (HFPNe). HFPNe can seamlessly handle discrete and continuous objects and events while keeping the model components themselves simple. With the feature and graphical model editor, biopathways can be modelled intuitively and simulated on GON. The subsequent output of the simulation results can be evaluated in customised views on GON Visualizer by writing an XML file. Additionally, GON provides a tool to transform biopathway models in KEGG and BioCyc to the GON XML files for modelling and simulation. The tool avoids a lot of tedious work by users, enabling them to focus on the biological model. PMID- 15130807 TI - Genomic Object Net: II. Modelling biopathways by hybrid functional Petri net with extension. AB - This paper demonstrates how to create an HFPNe (hybrid functional Petri net with extension) model, using the lac operon gene regulatory mechanism and glycolytic pathway as an example. Using this example, readers can then model other biopathways of interest. Simulations of the HFPNe model were performed using the software package Genomic Object Net. PMID- 15130808 TI - A protocol for the update of references to scientific literature in biological databases. AB - Entries in biological databases are usually linked to scientific references. To generate those links and to keep them up-to-date, database maintainers have to continuously scan the scientific literature to select references that are relevant for each single database entry. The continuous growth of both the corpus of scientific literature and the size of biological databases makes this task very hard. We present a protocol intended to assist the updating of an existing set of literature (abstract) links from a single database entry with new references. It consists of taking the set of MEDLINE neighbour references of the existing linked abstracts and evaluating their relevance according to the existing set of abstracts. To test the applicability of the algorithm, we did a simple benchmark of the system using the references associated with the entries of a protein domain database. Human experts found the references that the algorithm scored highly were more relevant to the database entry than those scored lowly, suggesting that the algorithm was useful. PMID- 15130809 TI - Proceedings of the New Zealand Bioinformatics Conference 2003. February, 2003. Wellington, New Zealand. PMID- 15130810 TI - Protein structure prediction and analysis as a tool for functional genomics. AB - Bioinformatic analyses of whole genome sequences highlight the problem of identifying the biochemical and cellular functions of the many gene products that are at present uncharacterised. Determination of their three-dimensional structures, either experimentally or by prediction, provides a powerful tool to address function, since it is at this level that biological activity is expressed. Here, we discuss the current approaches to protein structure prediction from sequence data, including the ab initio prediction of new folds, methods of fold recognition and comparative modelling based on homology. The value and limitations of such models are also explored. A major factor for the future will be the growth of the database of experimentally determined protein structures, through structural genomics projects. The prospects for this approach are also discussed, together with our experience in a pilot structural genomics project focused on proteins from Mycobacterium tuberculosis, the cause of tuberculosis (TB). PMID- 15130811 TI - Computational tools for analysing structural changes in proteins in solution. AB - Many important structural changes in proteins involve long-time dynamics, which are outside the timescale presently accessible by a straightforward integration of Newton's equations of motion. This problem is addressed with minimisation based algorithms, which are applied on possible reaction pathways using atomic detail models. For reasons of efficiency, an implicit treatment of solvent is imperative. We present the charge reparameterisation protocol, which is a method that approximates the interaction energies obtained by a numerical solution of the Poisson-Boltzmann equation. Furthermore, we present a number of methods that can be used to compute possible reaction pathways associated with a particular conformational change. Two of them, the self-penalty walk and the nudged elastic band method, define an objective function, which is minimised to find optimal paths. A third method, conjugate peak refinement, is a heuristic method, which finds minimum energy paths without the use of an explicit objective function. Finally, we discuss problems and limitations with these methods and give a perspective on future research. PMID- 15130812 TI - Visualisation and navigation methods for typed protein-protein interaction networks. AB - Protein-protein interactions form large and complex networks. Their visualisation can aid biologists in gaining new insights about the processes in cells and is, therefore, very useful for building sophisticated research tools. Often standard force-directed graph drawing algorithms are used for the visualisation of these networks. However, currently available visual interfaces to biological databases only show general interactions and cannot cope well with more complex networks with different types of interactions. This paper presents a new approach to the visual analysis of protein-protein interaction networks. It uses a combination of circular and force-directed graph drawing algorithms to compute visual representations of protein networks depending on the type of the selected interaction. Smooth transitions between subsequent drawings enable users to explore different functional clusters in these networks without getting lost in the entire network. The visualisation system has been tested with data from the BRITE database. PMID- 15130813 TI - The prion paradox: infection or polymerisation? AB - A weak but significant similarity was found between the prion protein (PrP) and some transcription factors and zinc-finger proteins. A possible interpretation of this similarity is that the PrP is a metal- (copper-) binding transcription factor and might behave like a Zn-finger protein, with the Cu2+ binding to its histidine and serine residues. Copper-binding could create intramolecular bridges in the PrPC (normal, cytoplasmic) molecule, but intermolecular bridges in the PrPSc (scrapie pathogenic) molecule. A molecular model of the Cu2+ -binding monomeric PrPC and the Cu2+-stabilised polymeric PrP Sc is presented here and named the 'cuprion model'. In this model, the PrPC has two idioforms. The stable, normal PrPC-idioform-I contains Cu2+ in -2His-Cu2+-2His- complexes and an intramolecular disulphide bridge. An unstable, transient form, PrPC-idioform-II, contains a -2His-Cu 2+-2Cys- complex, which destabilises the intramolecular disulphide bridge and makes the PrPC molecule highly reactive with other PrPC molecules. PMID- 15130814 TI - Gap mapping: a paradigm for aligning two sequences. AB - Pairwise sequence alignment is one of the most essential tools in comparative genomic sequence analysis. It is used to compare the sequences of genes and proteins with the aim of inferring structural, functional and evolutionary relationships. However, current 'mainstream' alignment algorithms have optimisation criteria based primarily on computational efficiency using parameters such as gap penalties, which are not biologically motivated. In addition, current alignment algorithms such as the Smith and Waterman technique provide a single alignment that could be sensitive to rather arbitrary choices in parameters such as gap penalties. This paper explores the range of properties resulting from posing the alignment problem more as a 'mapping gaps in sequences' exercise. We argue that this approach is intuitive and provides greater control over the number of gaps placed within an alignment. This type of approach was proposed by Sankoff (1972), but unfortunately has not received much attention. We report and discuss our findings by comparing this approach to other techniques using structurally confirmed aligned sequences from a benchmark alignment database. Interestingly, this approach consistently provides optimal and near optimal alignments and is thus a viable approach to sequence alignment. PMID- 15130815 TI - Complementary techniques of clustering and composite pattern analysis to Saccharomyces cerevisiae gene expression. AB - We present the combined application of clustering and composite pattern analysis to Saccharomyces cerevisiae gene expression and sequence data. The aim is to devise a system for the efficient investigation of the upstream regions associated with regulating the coordinated transcriptional response of genes via the location of cis-acting elements. The clustering algorithm presented allows clusters to overlap in light of the fact that many genes are regulated by multiple factors and/or are associated with multiple functions in the organism. The sequence clusters are analysed for shared monad and dyad patterns. These motifs are commonly regarded as putative cis-acting regulatory elements. Computationally, dyad patterns may be difficult and expensive to discover and verify if the sequences are very large. This has highlighted the need to obtain maximally--with regards to the pattern in question--'enriched' sets of sequences to uncover these motifs. We present overlapping clusters as an ideal basis for the discovery of conserved motifs. PMID- 15130816 TI - Detection of signals in mRNAs that influence translation. AB - Genome sequencing efforts mean that we now have extensive data from a wide range of organisms to study. Understanding the differing natures of the biology of these organisms is an important aim of genome analysis. We are interested in signals that affect translation of mRNAs. Some signals in the mRNA influence how efficiently it is translated into protein. Previous studies have indicated that many important signals are located around the initiation and termination codons. We have developed tools described here to extract the relevant sequence regions from GenBank. To create databases organised by species, or higher taxonomic groupings (eg planta), a program was developed to dynamically view and edit the taxonomy database. Data from relevant species were then extracted using our Genbank feature table parser. We analysed all available sequences, particularly those from complete genomes. Patterns were then identified using information theory. The software is available from http://transterm.otago.ac.nz. Patterns around the initiation codons for most of the organisms fall into two groups, containing the previously known Shine-Dalgarno and Kozaks efficiency signals. However, we have identified several organisms that appear to utilise novel systems. Our analysis indicates that some organisms with extremely high GC% genomes do not have a strong dependence on base pairing ribosome binding sites, as the complementary sequence is absent from many genes. PMID- 15130817 TI - Prediction of clinical behaviour and treatment for cancers. AB - Prediction of clinical behaviour and treatment for cancers is based on the integration of clinical and pathological parameters. Recent reports have demonstrated that gene expression profiling provides a powerful new approach for determining disease outcome. If clinical and microarray data each contain independent information then it should be possible to combine these datasets to gain more accurate prognostic information. Here, we have used existing clinical information and microarray data to generate a combined prognostic model for outcome prediction for diffuse large B-cell lymphoma (DLBCL). A prediction accuracy of 87.5% was achieved. This constitutes a significant improvement compared to the previously most accurate prognostic model with an accuracy of 77.6%. The model introduced here may be generally applicable to the combination of various types of molecular and clinical data for improving medical decision support systems and individualising patient care. PMID- 15130818 TI - Text mining of DNA sequence homology searches. AB - Primary tasks in analysis and annotation of expressed sequence tag (EST) datasets are to identify similarity among sequences by unsupervised clustering and assign putative function based on BLAST homology searches. We investigated the usefulness of text mining as a simple approach for further higher-level clustering of EST datasets using IBM Intelligent Miner for Text v2.3 tools. Agglomerative and k-means clustering tools were used to cluster BLASTx homology search documents from two onion EST datasets and optimised by pre-processing and pruning. Subjective evaluation confirmed that these tools provided biologically useful and complementary views of the two libraries, provided new insights into their composition and revealed clusters previously identified by human experts. We compared BLASTx textual clusters for two gene families with their DNA sequence based clusters and confirmed that these shared similar morphology. PMID- 15130819 TI - Identifying tissue-enriched gene expression in mouse tissues using the NIH UniGene database. AB - There is considerable interest in the gene expression profiles that underpin the phenotypes of cells and tissues. We have developed Bioperl scripts for mining the National Institutes of Health (NIH) UniGene databases to identify this tissue enriched gene expression. UniGene imports expressed sequence tags (ESTs) from the NIH dbEST database and clusters them by searching for sequence matches. In principle, each UniGene cluster represents the product(s) of a single transcriptional unit in the genome. This transcriptional unit can be expressed in a range of cell types, and UniGene clusters reflect these heterogeneous origins. UniGene clusters containing ESTs expressed predominantly or uniquely by one tissue will show a high proportion of ESTs from that tissue. Our Bioperl scripts parse the NIH UniGene data files as a starting point for an in-house UniGene database. Each UniGene cluster is then assessed for the total number of ESTs from a specified set of dbEST libraries and the total number of ESTs in the cluster. The ratio of the two gives a measure of enrichment. In this paper, we identify tissue-enriched gene expression in mouse pancreas, mammary gland and heart. Each tissue-enriched expression profile identifies genes that are recognisably characteristic of the respective tissue. It also identifies significant numbers of tissue-enhanced UniGenes that are derived from transcriptional units with no known function. These genes may play important and specialised functions in the tissue in question and offer targets for drug action. PMID- 15130820 TI - Ensemble machine learning on gene expression data for cancer classification. AB - Whole genome RNA expression studies permit systematic approaches to understanding the correlation between gene expression profiles to disease states or different developmental stages of a cell. Microarray analysis provides quantitative information about the complete transcription profile of cells that facilitate drug and therapeutics development, disease diagnosis, and understanding in the basic cell biology. One of the challenges in microarray analysis, especially in cancerous gene expression profiles, is to identify genes or groups of genes that are highly expressed in tumour cells but not in normal cells and vice versa. Previously, we have shown that ensemble machine learning consistently performs well in classifying biological data. In this paper, we focus on three different supervised machine learning techniques in cancer classification, namely C4.5 decision tree, and bagged and boosted decision trees. We have performed classification tasks on seven publicly available cancerous microarray data and compared the classification/prediction performance of these methods. We have observed that ensemble learning (bagged and boosted decision trees) often performs better than single decision trees in this classification task. PMID- 15130821 TI - Using ancestral sequences to uncover potential gene homologues. AB - Gene homologues between distantly related species can be difficult to identify. We test the idea that inferred ancestral sequences could aid in finding gene homologues. Ancestral sequences are inferred by aligning gene homologues on a known tree and estimating the most likely amino acid for each position at each node in that tree. BLAST(R) and HMMER are used separately and together with ancestral sequences to search the genome sequence databases of Encephalitozoon cuniculi, Entamoeba histolytica and Giardia lamblia for RNase P protein homologues. RNase P proteins (Pop4, Pop1, Pop5 and Rpp21) have been reported in humans and at least two other eukaryotic species but have yet to be identified in the above genomes. Using ancestral sequences reconstruction (ASR) for these proteins, we successfully identified putative homologues from E. cuniculi, Ent. histolytica and G. lamblia. In some cases, the use of ASR outperformed BLAST and HMMER. Overall, including ancestral sequences in searches with BLAST and/or HMMER was the most successful approach in the recovery of potential RNase P protein gene homologues, making this a useful technique in early homologue identification. PMID- 15130822 TI - Using serum albumin to infer vertebrate phylogenies. AB - The phylogenetic relationships of the two extant lobe-finned fish, coelacanth and lungfish, to tetrapods remain unresolved. In our study of serum albumin in lungfish we determined 101 residues of protein sequence from five peptides. The lungfish sequences were aligned with the albumin superfamily using the UPGMA based pileup or the progressive ALIGN programs. The tree construction methods employed were maximum parsimony, neighbour joining and TREE. The lungfish albumin sequence was compared with that of other species, and the resulting trees indicated a close relationship of lungfish to tetrapods, in line with the bulk of current phylogenetic evidence. PMID- 15130823 TI - Support vector machine applications in bioinformatics. AB - The support vector machine (SVM) approach represents a data-driven method for solving classification tasks. It has been shown to produce lower prediction error compared to classifiers based on other methods like artificial neural networks, especially when large numbers of features are considered for sample description. In this review, the theory and main principles of the SVM approach are outlined, and successful applications in traditional areas of bioinformatics research are described. Current developments in techniques related to the SVM approach are reviewed which might become relevant for future functional genomics and chemogenomics projects. In a comparative study, we developed neural network and SVM models to identify small organic molecules that potentially modulate the function of G-protein coupled receptors. The SVM system was able to correctly classify approximately 90% of the compounds in a cross-validation study yielding a Matthews correlation coefficient of 0.78. This classifier can be used for fast filtering of compound libraries in virtual screening applications. PMID- 15130824 TI - Integrative modelling of gene expression and cell metabolism. AB - The goal of modelling biochemical networks is to understand the system's behaviour (dynamics and control) of these networks in terms of the properties of the individual molecules. Most modelling approaches have dealt with either gene expression or cell metabolism. In light of the widespread use of robotic technologies in laboratories and improved computational power, it is now time to incorporate information from all biochemical levels--gene expression, protein interactions and metabolism--into integrated models. Here, we review the literature on modelling gene expression with cell metabolism. At the end we describe some analytic methods to deal with these systems. PMID- 15130825 TI - Gene duplication, the evolution of novel gene functions, and detecting functional divergence of duplicates in silico. AB - Duplication of genes increases the amount of genetic material on which evolution can work and has been considered of major importance for the development of biological novelties or to explain important transitions that have occurred during biological evolution. Recently, much research has been devoted to the study of the evolutionary and functional divergence of duplicated genes. Since the majority of genes are part of gene families, there is considerable interest in predicting differences in function between duplicates and assessing the functional redundancy of genes within gene families. In this review, we discuss the strengths and limitations of both older and novel approaches to investigate the evolution of duplicated genes in silico. PMID- 15130826 TI - DNA sequence analysis linguistic tools: contrast vocabularies, compositional spectra and linguistic complexity. AB - This is a review of the methods based on counting oligomers in nucleotide and amino acid sequences. Such methods are analogous to the formal linguistic analysis of human texts. This review includes methods based on the calculation of observed occurrences (frequencies) of oligomers and their distribution, as well as those based on deviations between the observed and the expected occurrences (contrast words, genome signatures) in biological sequences. Both types of methods have a wide range of sensitivity and can identify homologous as well as functionally and taxonomically related sequences. PMID- 15130827 TI - Paper2sequences: retrieval of sequences listed in a publication. AB - Our web-based tool simplifies the often laborious procedure of retrieving a set of biosequences in a publication or webpage. As a front-end to the Bioperl toolkit, it accepts as an input a list of identifiers. They are specified in an ASCII table (copy-pasted from the publication's PDF or HTML page) and give rise to queries in multiple databases for the protein/nucleic acid data specified. Currently, GenBank, PIR (Protein Information Resource) and Swiss-Prot are supported. For any sequence accession code listed, the database can be specified and, if retrieval fails, automatic lookup for the same code in other databases can be requested. Sequence length information (if specified) and heuristic rules are used to drive the lookup if multiple protein coding sequences (CDS) are part of a single accession. Warnings are issued in cases of ambiguities and inconsistencies. An advanced option enables the user to format the output in whatever format they wish. PMID- 15130828 TI - A library of efficient bioinformatics algorithms. AB - In this paper we review some of the existing projects available in the bioinformatics field for facilitating the development of programs, but for which minimising the running time is not of primary importance. We point out the advantages of open source libraries for such tasks and we discuss some of the open source licenses available. Finally, we present the project ALiBio, which is aimed at facilitating the development of efficient programs in bioinformatics. PMID- 15130830 TI - Protein function from sequence and structure data. AB - With the large amount of genomics and proteomics data that we are confronted with, computational support for the elucidation of protein function becomes more and more pressing. Many different kinds of biological data harbour signals of protein function, but these signals are often concealed. Computational methods that use protein sequence and structure data can be used for discovering these signals. They provide information that can substantially speed up experimental function elucidation. In this review we concentrate on such methods. PMID- 15130831 TI - Innovation from reduction: gene loss, domain loss and sequence divergence in genome evolution. AB - Analyses of genome sequences have revealed a surprisingly variable distribution of genes, reflecting the generation of novel genes, lateral gene transfer and gene loss. The impact of gene loss on organisms has been difficult to examine, but the loss of protein coding genes, the loss of domains within proteins and the divergence of genes have made surprising contributions to the differences among organisms. This paper reviews surveys of gene loss and divergence in fungal and archaeal genomes that indicate suites of functionally related genes tend to undergo loss and divergence. Instances of fungal gene loss highlighted here suggest that specific cellular systems have changed, such as Ca 2+ biology in Saccharomyces cerevisiae and peroxisome function in Schizosaccharomyces pombe. Analyses of loss and divergence can provide specific predictions regarding protein-protein interactions, and the relationship between networks of protein interactions and loss may form a part of a parametric model of genome evolution. PMID- 15130832 TI - Microarray data clustering based on temporal variation: FCV with TSD preclustering. AB - The aim of this paper is to present a new clustering algorithm for short time series gene expression data that is able to characterise temporal relations in the clustering environment (ie data-space), which is not achieved by other conventional clustering algorithms such as k -means or hierarchical clustering. The algorithm called fuzzy c -varieties clustering with transitional state discrimination preclustering (FCV-TSD) is a two-step approach which identifies groups of points ordered in a line configuration in particular locations and orientations of the data-space that correspond to similar expressions in the time domain. We present the validation of the algorithm with both artificial and real experimental datasets, where k -means and random clustering are used for comparison. The performance was evaluated with a measure for internal cluster correlation and the geometrical properties of the clusters, showing that the FCV TSD algorithm had better performance than the k -means algorithm on both datasets. PMID- 15130833 TI - Comparison of responses by bacteriophages and bacteria to pressures on the base composition of open reading frames. AB - Differences in the base composition of genomes can occur because of GC pressure, purine-loading pressure (AG pressure) and RNY pressure, for which there are possible functional explanations, and because of the more abstract pressures exerted by individual bases. The graphical approach of Muto and Osawa was used to analyse how bacteriophages and bacteria balance potentially conflicting pressures on their genomes. Phages generally respond to AG pressure by increasing A while keeping T constant, and by decreasing C while keeping G constant. In contrast, bacteria generally increase both A and T, the former more so, and decrease both G and C, the latter more so. These differences largely occur at third codon positions, which are more responsive than first and second codon positions to AG pressure and GC pressure. Phages respond to AG pressure more in the third codon position than bacteria, whereas bacteria respond more in the first codon position than phages. Conversely, bacteria respond to GC pressure more in the third codon position than phages, whereas phages respond more in the first codon position than bacteria. As GC pressure increases, A is traded for C and AG pressure decreases; first and second codon positions, having more A than T, are most responsive to this negative effect of increased GC pressure; third positions either do not respond (phages) or respond weakly (bacteria). In a set of 48 phage host pairs, degrees of purine loading were less correlated between phage and host than were GC percentages. These results suggest that pressures on conventional and genome phenotypes operate differentially in phages and bacteria, generating both general differences in base composition and specific differences characteristic of particular phage-host pairs. The reciprocal relationship between GC pressure and AG pressure implies that effects attributed to GC pressure may actually be due to AG pressure, and vice versa. PMID- 15130834 TI - MHCPred: bringing a quantitative dimension to the online prediction of MHC binding. AB - The accurate prediction of T cell epitopes is one of the key aspirations of immunoinformatics. We have developed a partial least squares-based, robust multivariate statistical method for the quantitative prediction of peptide binding to major histocompatibility complexes (MHCs), the principal checkpoint on the antigen presentation pathway. As a service to the immunobiology community, we have made a Perl implementation of the method available as a World Wide Web server. PMID- 15130835 TI - Immunoinformatics and the prediction of immunogenicity. AB - Immunoinformatics is the application of informatics techniques to molecules of the immune system. One of the key goals of immunoinformatics is the development of computer aided vaccine design (CAVD), or computational vaccinology, and its application to the search for new vaccines. Key to solving this challenge is the prediction of immunogenicity, be that at the level of epitope, subunit vaccine or attenuated pathogen. This paper reviews the current state of play in the prediction of immunogenicity and focuses on well developed methods for the prediction of peptide binding affinity to major histocompatibility complexes, which are the necessary preliminary to the in silico identification of T cell epitopes. PMID- 15130836 TI - Comparative modelling: an essential methodology for protein structure prediction in the post-genomic era. AB - The gap between the number of protein sequences and protein structures is increasing rapidly, exacerbated by the completion of numerous genome projects now flooding into public databases. To fill this gap, comparative protein modelling is widely considered the most accurate technique for predicting the three dimensional shape of proteins. High-throughput, automatic protein modelling should considerably increase our access to protein structures other than those determined by experimental techniques such as X-ray crystallography and NMR (nuclear magnetic resonance) spectroscopy. The uses for these complete three dimensional models are growing rapidly, ranging from guiding site-directed mutagenesis experiments to protein-protein interaction predictions. In recognition of this, a number of very useful comparative modelling servers have begun to emerge on the Web. Molecular biologists now have a powerful web-based toolkit to construct models, assess their accuracy, and use them to explain and predict experiments. There is, however, still much to do by those engaged in algorithmic development if comparative modelling is to compete on an equal footing with experimental protein structure determination techniques. PMID- 15130837 TI - Artificial intelligence techniques for bioinformatics. AB - This review provides an overview of the ways in which techniques from artificial intelligence (AI) can be usefully employed in bioinformatics, both for modelling biological data and for making new discoveries. The paper covers three techniques: symbolic machine learning approaches (nearest neighbour and identification tree techniques), artificial neural networks and genetic algorithms. Each technique is introduced and supported with examples taken from the bioinformatics literature. These examples include folding prediction, viral protease cleavage prediction, classification, multiple sequence alignment and microarray gene expression analysis. PMID- 15130838 TI - 3D-Hit: fast structural comparison of proteins. AB - 3D-Hit is a fast scanning method for detecting structural similarities between proteins. The algorithm is based on a hashing function, which decomposes proteins into segments of 13 residues. The scanning procedures start with assigning a set of similar segments from the database to each segment in the query protein. These initial hits are expanded by two iterations of structural superposition of larger segments of 99 and 299 residues. The method generates an alignment for the query protein by concatenating partial structural alignments. PMID- 15130839 TI - Consensus sequence Zen. AB - Consensus sequences are widely used in molecular biology but they have many flaws. As a result, binding sites of proteins and other molecules are missed during studies of genetic sequences and important biological effects cannot be seen. Information theory provides a mathematically robust way to avoid consensus sequences. Instead of using consensus sequences, sequence conservation can be quantitatively presented in bits of information by using sequence logo graphics to represent the average of a set of sites, and sequence walker graphics to represent individual sites. PMID- 15130840 TI - Perspectives on protein evolution from simple exact models. AB - Understanding the evolution of biopolymers is important to rationalise the directed and undirected design of functional molecules. Large scale experiments or detailed computational studies are often impractical. Therefore, simple model systems, such as RNA secondary structure and lattice proteins have been adapted to study general statistical and topological features of genotype (sequence) to phenotype (structure) maps. We review findings from such models that address aspects of thermodynamic and mutational robustness, neutral evolution and recombination of proteins. We compare various modelling approaches, and discuss their generality, parameter dependency and experimental verifications of their predictions. The most striking observation is the universal emergence of neutral nets--sets of phenotypically identical genotypes that are interconnected by series of point mutations. However, fast adaptation by point mutations appears to be problematic for proteins. This may explain why proteins appear to be more specific while RNA is rather versatile. This could even be the reason why RNA had to evolve before proteins. Similar principles of biological organisation are reflected in sequence and structure databases of real proteins. Insights gained from modelling are useful for designing more efficient database organisation and search strategies. PMID- 15130841 TI - rRNA-like sequences in human mRNAs. AB - Many eukaryotic transcripts have been reported to contain sequences similar or complementary to ribosomal RNAs. Past computational and experimental analyses have suggested a possible functional role of such rRNA-like sequences in the regulation of translation. Here, we analysed the occurrence and abundance of rRNA like sequences in human transcripts. Using the wealth of data from genome and EST sequences, our study indicates that mRNA sequences with long highly similar rRNA like sequences are most likely artifacts. In contrast, analysis of the occurrence of 9-nt sequences covering the entire direct and complementary 18S human rRNA sequence, revealed that many of them are significantly over- or underrepresented, suggesting their possible involvement in post-transcriptional regulation of gene expression. PMID- 15130842 TI - TAD: a web interface and database for tissue microarrays. AB - Tissue microarrays are increasingly important tools that bring high-throughput technology to traditional pathology laboratories. In many cases, each spot on a tissue microarray is scored by a skilled pathologist and recorded manually. TAD consists of an Active Server Page web interface to a relational database that automates recording scores and linking them with clinical data for future interpretation. TAD is an open source application that can be installed locally. PMID- 15130843 TI - Spectronet: a package for computing spectra and median networks. AB - Spectronet is a package that uses various methods for exploring and visualising complex evolutionary signals. Given an alignment in NEXUS format, the package works by computing a collection of weighted splits or bipartitions of the taxa and then allows the user to interactively analyse the resulting collection using tools such as Lento-plots and median networks. The package is highly interactive and available for PCs. PMID- 15130844 TI - MProbe: computer aided probe design for oligonucleotide microarrays. AB - The present work describes a complete probe design software system for oligonucleotide microarrays based on Kane's research on probe sensitivity and specificity (Kane's rule). Combining Kane's rule and traditional criteria for probe design we constructed MProbe, the software system for oligonucleotide microarrays using Java. The general criteria for probe design are: (1) probes may have different lengths that range from 20 to 100 bases; (2) they should have a similar melting temperature (Tm) or GC content; (3) they should not contain stable secondary structures; and (4) they abide by Kane's rule. PMID- 15130845 TI - An update on molecular genetic studies of human personality traits. AB - Personality is a complex phenotype and people differ considerably when they are evaluated by self-report questionnaires. There is convincing evidence from twin studies that basic personality dimensions in men and women have a considerable genetic component. However, only recently have common genetic polymorphisms been associated with particular personality traits, especially the dopamine D4 receptor with novelty seeking and the serotonin transporter with anxiety-related traits or neuroticism. The current review examines progress in the past few years in molecular personality genetics and focuses on the reasons for difficulties in replicating first findings as well as the prospects for future studies in this area. The molecular genetic structure of human personality is worth studying both for its intrinsic interest in helping us to understand individual differences in human behaviour and the light it will shed on more complex behavioural disorders that are likely to partially share some common genetic variants. PMID- 15130846 TI - EYE on bioinformatics: dissecting complex disease traits in silico. AB - Bioinformatics has provided an unprecedented power and resource for us to decipher the enigma of complex diseases. It can reveal otherwise promiscuous information from the tremendous amount of data generated by the new, powerful and high-throughput technologies of genomics and proteomics. In this paper, we review the cutting edge developments in complex disease trait mapping, databases, computational gene recognition, gene function prediction, pathway reconstruction and disease classification by expression profiling, and computational modelling of living systems. Integration of all this knowledge and the different technologies, alongside cooperation between experts from different fields, will enhance our understanding of the molecular and mechanistic abnormalities in disease state, and greatly assist the rational development of effective therapies. PMID- 15130848 TI - Predicting functional linkages from gene fusions with confidence. AB - Pairs of genes that function together in a pathway or cellular system can sometimes be found fused together in another organism as a Rosetta Stone protein- a fusion protein whose separate domains are homologous to the two functionally related proteins. The finding of such a Rosetta Stone protein allows the prediction of a functional linkage between the component proteins. The significance of these deduced functional linkages, however, varies depending on the prevalence of each of the two domains. Here, we develop a statistical measure for the significance of predicted functional linkages, and test this measure for proteins of E. coli on a functional benchmark based on the KEGG database. By applying this statistical measure, proteins can be linked with over 70% accuracy. Using the Rosetta Stone method and this scoring scheme, we find all significant functional linkages for proteins of E. coli, P. horikshii and S. cerevisiae, and measure the extent of the resulting protein networks. PMID- 15130849 TI - APBioNet: the Asia-Pacific regional consortium for bioinformatics. AB - Bioinformatics and computational biology, along with the related fields of genomics, proteomics, functional genomics and systems biology are new wave scientific disciplines that harness composite computational power across networks to advance biological knowledge at the most basic level and to direct traditional laboratory-based research efforts in the biomedical sciences. 'Fostering the growth of bioinformatics and allied disciplines in the Asia-Pacific region' is the motto of the first regional bioinformatics society, the Asia-Pacific Bioinformatics Network (APBioNet). APBioNet addresses the issues of hardware, software, databases and networks pertaining to bioinformatics, with the additional layer of pertinent education, training and research. Recent milestones achieved include hosting an international bioinformatics symposium in Asia and setting up large-scale regional grid-computing projects. PMID- 15130850 TI - ScanProsite: a reference implementation of a PROSITE scanning tool. AB - Many different software tools are available publicly to scan the PROSITE database of protein families. However, none of them, to our knowledge, wholly implements the PROSITE syntax, or satisfies all the rules for scanning a pattern against a sequence. We hereby propose a strict definition of how a PROSITE pattern is to be scanned against a sequence, and provide a reference implementation of a tool to scan PROSITE patterns, rules and profiles against protein sequences. PMID- 15130847 TI - Genomic biodiversity, phylogenetics and coevolution in proteins. AB - Comprehensive sampling of genomic biodiversity is fast becoming a reality for some genomic regions and complete organelle genomes. Genomic biodiversity is defined as large genomic sequences from many species, and here some recent work is reviewed that demonstrates the potential benefits of genomic biodiversity for molecular evolutionary analysis and phylogenetic reconstruction. This work shows that using likelihood-based approaches, taxon addition can dramatically improve phylogenetic reconstruction. Features or dynamics of the evolutionary process are much more easily inferred with large numbers of taxa, and large numbers are essential for discriminating differences in evolutionary patterns between sites. Accurate prediction of site-specific patterns can improve phylogenetic reconstruction by an amount equivalent to quadrupling sequence length. Genomic biodiversity is particularly central to research relating patterns of evolution, adaptation and coevolution to structural and functional features of proteins. Research on detecting coevolution between amino acid residues in proteins demonstrates a clear need for much greater numbers of closely related taxa to better discriminate site-specific patterns of interaction, and to allow more detailed analysis of coevolutionary interactions between subunits in protein complexes. It is argued that parsing out coevolutionary and other context dependent substitution probabilities is essential for discriminating between coevolution and adaptation, and for more realistically modelling the evolution of proteins. Also reviewed is research that argues for increasing the efficiency of acquiring genomic biodiversity, and suggests that this might be done by simultaneously shotgun cloning and sequencing genomic mixtures from many species. Increased efficiency is a prerequisite if genomic biodiversity levels are to rapidly increase by orders of magnitude, and thus lead to dramatically improved understanding of interactions between protein structure, function and sequence evolution. PMID- 15130852 TI - A hierarchical model of HIV-1 protease drug resistance. AB - A hierarchical model of HIV-1 drug therapy may be used to evaluate new inhibitors and test new treatment strategies that address the problem of drug resistance. The model includes an atomic representation of drug-protease interaction, evaluation of viral fitness based on cleavage of polyprotein substrates during viral maturation, evolutionary modelling of drug resistance mutations in the face of selection pressures by drug and a mathematical description of viral population dynamics in infected individuals. These techniques have been used for the design of resistance-evading inhibitors by computational coevolution techniques and for the optimisation of existing protease inhibitors for improving their robustness in the face of resistance mutation. PMID- 15130853 TI - Distributed computing in bioinformatics. AB - This paper provides an overview of methods and current applications of distributed computing in bioinformatics. Distributed computing is a strategy of dividing a large workload among multiple computers to reduce processing time, or to make use of resources such as programs and databases that are not available on all computers. Participating computers may be connected either through a local high-speed network or through the Internet. PMID- 15130854 TI - State-of-the-art in membrane protein prediction. AB - Membrane proteins are crucial for many biological functions and have become attractive targets for pharmacological agents. About 10%-30% of all proteins contain membrane-spanning helices. Despite recent successes, high-resolution structures for membrane proteins remain exceptional. The gap between known sequences and known structures calls for finding solutions through bioinformatics. While many methods predict membrane helices, very few predict membrane strands. The good news is that most methods for helical membrane proteins are available and are more often right than wrong. The best current prediction methods appear to correctly predict all membrane helices for about 50% 70% of all proteins, and to falsely predict membrane helices for about 10% of all globular proteins. The bad news is that developers have seriously overestimated the accuracy of their methods. In particular, while simple hydrophobicity scales identify many membrane helices, they frequently and incorrectly predict membrane helices in globular proteins. Additionally, all methods tend to confuse signal peptides with membrane helices. Nonetheless, wet-lab biologists can reach into an impressive toolbox for membrane protein predictions. However, the computational biologists will have to improve their methods considerably before they reach the levels of accuracy they claim. PMID- 15130855 TI - Virus bioinformatics: databases and recent applications. AB - Bioinformatics is now used as an umbrella term for almost all aspects of computational biology. Bioinformatics research will have an impact on all of biology, and virology is not immune from these research methods. Although virology has been slower to embrace bioinformatics this is now changing, particularly in the areas of viral sequences databasing and the systematic identification of viral and host homologous proteins. Here we will review some of these recent advances focusing mainly on the herpesvirus. PMID- 15130856 TI - The hand, foot and mouth disease virus capsid: sequence analysis and prediction of antigenic sites from homology modelling. AB - Enterovirus 71 (EV71) is the most common aetiological agent detected in cases of hand, foot and mouth disease (HFMD) resulting in incidences of neurological complications and fatality in recent years. A comparison of the capsid proteins implicated in the pathogenicity of the fatal and non-fatal strains of EV71, reveals a high degree of homology (93%-100% identity). To facilitate diagnostic immunoassays and vaccine development, a consensus structural model for the EV71 coat protein has been developed based primarily on the homologous structure of the bovine enterovirus. The overall architecture of the virion closely resembles those of related icosahedral picornaviruses. Detailed atomic modelling of the fatal 5865/SIN/00009 strain has been carried out, and the functional regions (known and predicted) from closely related viruses mapped onto the surface of the predicted structure. From the model, we have identified two putative immunogenic regions, one of which is unique to EV71. The hydrophobic pocket within VP1, found in bovine enterovirus, poliovirus and rhinovirus, is also conserved in EV71. PMID- 15130857 TI - Towards a bioinformatics network for Latin America and the Caribbean (LACBioNet). AB - Bioinformatics is increasingly recognised as a crucial field for research and development in the biological sciences, and forms an integral part of genomics, proteomics and modern biotechnology. Worldwide participation is important, and scientists in developing countries can contribute to this field. Regional networks for bioinformatics are highly beneficial for capacity strengthening and cooperation, and for establishing productive interactions between scientists in the fields of biological and informatics sciences. Such a network (LACBioNet) is being organised for Latin America and the Caribbean. Its immediate goals include the organisation and extension of nodes and services, information and communication, research and development in different specialty fields of bioinformatics, and training and human resource development. PMID- 15130858 TI - Assertiveness with physicians: does it predict mammography use?. AB - In a prior study we found that women's self-reported assertiveness with their healthcare providers was associated with their use of mammography in a population based cross-sectional sample of women. Women who reported being more assertive, by repeating information if they felt their doctor didn't hear them, asking their doctor to explain information they didn't understand, or reminding their doctor about screening tests, were more likely to have received a mammogram recently than those who reported being less assertive. Here we examined how women's self reports of assertiveness predicted their use of mammography three years later. We examined this using a population-based sample of 781 women living in rural Washington State who were participating in a trial of mammography promotion. We found that assertive women were younger on average than less assertive women, but that even after controlling for age, education, income, and marital status, women who reported being assertive with their doctor in 1994 were more likely to receive regular mammograms in the next three years than those who did not (OR 2.1; CI 1.5, 2.9). If future studies also suggest that assertiveness predicts use of mammography or other preventive healthcare services, it would be valuable to examine the promotion of assertiveness as a means of improving public health. PMID- 15130859 TI - Cervical cancer screening among Latinas: the importance of referral and participation in parallel cancer screening behaviors. AB - Low cancer screening participation among medically underserved Latinas is largely due to lack of active referral to screening procedures by health care providers. We explored how physicians' referral and instruction on parallel screening procedures discriminates Latinas' cervical cancer screening practices in the context of relevant variables such as sociodemographic characteristics, health insurance, history of cancer, and level of acculturation. Of 153 women surveyed, 100 were compliant with yearly Pap smear while 53 were not compliant. Discriminant function analysis revealed that health care provider interventions and parallel breast cancer screening behaviors were significant discriminators between women who obtained a Pap smear within a year and those who were less compliant. A change in public health policy that facilitates to medically underserved Latinas access to reliable sources of health care referrals and services might increase their regular use of cervical cancer screening, which could potentially result in a reduction in cancer treatment costs and in lives lost to cervical cancer among these women. PMID- 15130860 TI - Measuring job stress and family stress in Chinese working women: a validation study focusing on blood pressure and psychosomatic symptoms. AB - Psychometric properties of a questionnaire measuring psychosocial work-related stress in terms of effort-reward imbalance and a short family stress scale were examined in a population sample of 421 working women from four work sites in Beijing, China. The internal consistency of the scales was satisfactory, and the theoretically postulated structure of scales of the work stress questionnaire was replicated. The criterion validity of the scales was tested using psychosomatic symptoms and blood pressure. Combined exposure to work and family- related stress was associated with an adjusted mean 6.4 mmHg increase in systolic blood pressure. Recurrent sleeping problems were also associated with the two stress measures. In conclusion, standardized measures of psychosocial stress in terms of effort-reward imbalance and of family stress can be used in occupational health research in China, with particular relevance for working women. PMID- 15130861 TI - After the fall: women's views of fractures in relation to bone health at midlife. AB - Past research has established the link between low energy fractures and the risk for future fractures. These fractures are potential markers for investigation of bone health, and may be precursors for osteoporosis. In spite of its significant public health burden, including burden of illness and economic costs, many individuals are not aware of the risk factors for and consequences of osteoporosis. This is a study of women aged 40 and older who experienced low energy fractures (e.g., from non-trauma sources and falls from no higher than standing height). We gathered data, using focus group interviews, about their experiences and understanding of the fractures in relation to bone health. Women often attributed the fractures to particular situations and external events (e.g., slipping on ice, tripping on uneven ground), and viewed the fractures as accidents. Women often felt that others are at risk for poor bone health, but believed that they themselves are different from those really at risk. Although the fractures are potential triggers for preventive efforts, few women connected their fracture to future risk. What is perceived by women (and others) as random and an accident is often a predictable event if underlying risk factors are identified. Only when there is more awareness of poor bone health as a disease process and fractures as markers for bone fragility will women, men and health care providers take action to prevent future fractures and established bone disease. PMID- 15130862 TI - Effects of home- and university-based programs on physical self-perception in mothers and daughters. AB - The primary purpose of this study was to compare two mother-daughter interventions designed to increase fitness and activity and improve physical self perception (PSP).A secondary purpose was to test two of the proposed mechanisms for using physical activity to enhance PSP. Twenty mother-daughter pairs (n = 40) were randomly assigned to a university-based (UB) or home-based (HB) group. Initially, both groups received classroom training designed to improve PSP. Then, the UB group met three times per week, whereas the HB group received information concerning home-based activities. The Fitnessgram activity questionnaire and fitness test battery were used to measure physical activity and fitness level. The Physical Self-Perception Profile (PSPP) was used to detect changes in PSP. Separate 2 (group) x 2 (age) x 2 (time) ANOVAs with repeated measures on the last factor were conducted to examine changes in PSP, physical activity, and fitness. Correlations between physical activity, fitness, and PSP were examined to test the two mechanisms. Perceived sports competence and body attractiveness improved in UB and HB mothers and daughters (p =.002 and.005, respectively). UB and HB mothers and daughters increased their participation in aerobic (p =.000 and.015), muscular strength (p =.001 and.001), and flexibility (p =.000 and.000) activities. Support was not provided for the proposed mechanisms for the relationship between physical activity and PSP. These interventions demonstrate the potential to foster positive short-term changes in PSP and physical activity in mothers and daughters. More research is needed to test mechanisms explaining the relationship between activity and PSP. PMID- 15130863 TI - An exploratory study of the health problems, stigmatization, life satisfaction, and literacy skills of urban, street-level sex workers. AB - An exploratory study of 26 female urban, street-level sex workers was conducted to gather information about their health problems, feelings of stigmatizations, satisfaction with life, and literacy skills. Each woman completed the health questionnaire, Stigmatization Scale, Satisfaction with Life Scale, and the Rapid Estimate of Adult Literacy in Medicine. Twenty-one women had acute or chronic health problems; only eleven sought health care. Literacy scores revealed 7th to 8th grade reading levels. Feelings of stigmatization varied from low to high and all the women were found to be dissatisfied with their lives. Further research needs to address how these factors affect their use of health care and outreach services. PMID- 15130864 TI - Prism aftereffects disrupt interlimb rhythmic coordination. AB - The authors examined effects of prism-induced proprioceptive aftereffects on coordination of 95 participants and compared interlimb rhythmic coordination performed before versus after exposure to prisms of varying optical displacements. The observed steady states of relative phase for postprism exposure coordination were shifted by a small but significant amount, but not across all prism conditions. Phase-shift direction was not specific to the direction of optical displacement and was not, across all conditions, proportional to the magnitude of optical displacement. Prism exposure was associated with increased relative phase variability for all prism conditions. A no-prism control group showed no changes in interlimb rhythmic coordination. The results suggest that prism-induced proprioceptive aftereffects have general, disruptive effects on interlimb rhythmic coordination. PMID- 15130865 TI - Effect of preparation on dual-task performance in postural control. AB - The authors applied an overlapping-task design to study the interaction between postural control and cognitive task processes in young (n = 10) and older (n = 10) adults. A rapid destabilizing floor translation was followed at specific time intervals by a simple auditory reaction time (RT) task. The translations were preceded by either an informational cue or no cue. Interference between postural task demands and the RT task was found only in the first 50 ms. Cueing also had an effect on both the onset of the postural recovery response and RT performance. The results suggest (a) only a brief interference between postural and cognitive processing demands in relatively easy tasks, (b) competition for a common central mechanism, possibly a response-selection mechanism, and (c) no differential impact of aging on that interaction. PMID- 15130866 TI - An emerging postural response: is control of the hip possible in the newly walking child? AB - Previous researchers have proposed that because of their slow muscle-response latencies, 1- to 2-year-old children are unable to control hip-dominant postural responses when responding to balance threats (G. McCollum & T. Leen, 1989). To test that proposition, the authors exposed 41 children to backward support surface translations and recorded muscle activations and movement kinematics. Children between 10 months and 10 years of age stood on a platform that was unexpectedly moved. Passive hip-dominant responses were observed among the least experienced walkers. In contrast, older children produced an active response, signified by higher levels of abdominal and quadriceps muscle activity and accompanied by larger hip flexor torques. Greater success in withstanding large magnitudes of perturbations were associated with actively generated hip responses. PMID- 15130867 TI - Visual control of manual aiming movements in 6- to 10-year-old children and adults. AB - Recent results indicate that adults modulate their initial movement impulse toward a stationary visual target by processing visual afferent information. The authors investigated whether the mechanisms responsible for those modulations are already in place in young children or develop as the children grow older. Adults (n = 10) and 6-, 8- and 10-year-old children (ns = 6, 7, and 7, respectively) performed a video-aiming task while vision of the cursor they were moving was (acquisition) or was not (transfer) visible. The results indicated that within participant variability of the initial impulse trajectory of the children's aiming movement leveled-off in acquisition between peak extent deceleration and the end of the initial impulse, whereas it increased linearly as movement unfolded in transfer. The results also indicated that children modulate their initial movement impulse when visual afferent information is available, although to a lesser extent than adults do, and strongly imply that contrary to past suggestions, the initial impulse of an aiming movement is not ballistic. PMID- 15130868 TI - Bimanual coordination dynamics in poststroke hemiparetics. AB - Poststroke hemiparetic individuals (n = 9) and a control group (n = 9) completed a frequency-scaled circle-drawing task in unimanual and bimanual conditions. Measures of intralimb spatial and temporal task accuracy and interlimb coordination parameters were analyzed. Significant reductions in task performance were seen in both limbs of the patients and controls with the introduction of bimanual movement. Spatial performance parameters suggested that the 2 groups focused on different hands during bimanual conditions. In the controls, interlimb coordination variables indicated predictable hand dominance effects, whereas in the patient group, dominance was influenced by the side of impairment and prior handedness of the individual. Therefore, in this particular bimanual task, performance improvements in the hemiplegic side could not be elicited. Intrinsic coupling asymmetries between the hands can be altered by unilateral motor deficits. PMID- 15130869 TI - Reciprocal influences of attentional focus on postural and suprapostural task performance. AB - The authors examined the influence that attentional focus on either a postural or a suprapostural task had on the performance of each task. Participants (N = 32) stood on an inflated rubber disk and held a pole horizontally. All participants performed under 4 attentional focus conditions: external (disk) or internal (feet) focus on the postural task, and external (pole) or internal (hands) focus on the suprapostural task. Compared with internal focuses, external focuses on either task resulted in similar and reduced postural sway. Response frequency on each task increased when participants focused on the respective task. Finally, an external focus on either task produced higher frequencies of responding on the suprapostural task. The authors conclude that suprapostural task goals have a stronger influence on postural control than vice versa, reflecting the propensity of the motor system to optimize control processes on the basis of the desired movement effect. PMID- 15130870 TI - Effects of response priming and inhibition on movement planning and execution. AB - The authors used a precueing method to examine the effects of response priming and inhibition on goal-directed action. Participants (N = 18) completed aiming movements to 1 of 2 locations following predictive (80% cued), nonpredictive (50% cued), and antipredictive (20% cued) precues at 1 of the 2 possible target locations. Consistent with previous research, participants responded more quickly to targets at cued locations than to targets at uncued locations in the 80% condition, and more quickly to targets presented at the uncued than to those presented at cued locations in the 50% and 20% conditions. As predicted by models of action-centered selective attention, movement trajectories deviated away from the cued location in the 50% condition. Movement trajectories were also altered in the 80% and the 20% conditions. Movements directed to the uncued location deviated away from the cued location in the 80% condition, whereas movements directed to the cued location deviated away from the uncued location in the 20% condition. The authors explain the latter trajectory results as a strategy of overcompensation. PMID- 15130871 TI - Challenge point: a framework for conceptualizing the effects of various practice conditions in motor learning. AB - The authors describe the effects of practice conditions in motor learning (e.g., contextual interference, knowledge of results) within the constraints of 2 experimental variables: skill level and task difficulty. They use a research framework to conceptualize the interaction of those variables on the basis of concepts from information theory and information processing. The fundamental idea is that motor tasks represent different challenges for performers of different abilities. The authors propose that learning is related to the information arising from performance, which should be optimized along functions relating the difficulty of the task to the skill level of the performer. Specific testable hypotheses arising from the framework are also described. PMID- 15130872 TI - Beyond curve fitting? Comment on Liu, Mayer-Kress, and Newell (2003). AB - Y.-T. Liu, G. Mayer-Kress, and K. M. Newell (2003) fit learning curves to movement time data and suggested 2 new methods for analyzing learning. They claimed that the methods go "beyond curve fitting." However, in neither their curve fitting nor their new methods is measurement noise accounted for, and therefore they produce inefficient and biased results. Using the data of Liu et al., in which variance caused by learning is small relative to the level of noise for most participants, the present authors demonstrate those problems and provide better alternatives that are more noise tolerant, more powerful, and go beyond curve fitting without displaying the extreme bias produced by the methods of Liu et al. PMID- 15130874 TI - Glutamine regulates Caco-2 cell tight junction proteins. AB - Intestinal epithelial tight junction (TJ) barrier dysfunction may lead to inflammation and mucosal injury. Glutamine (GLN) plays a role in maintenance of intestinal barrier function in various animal models and critically ill humans. Recent evidence from intestinal cell monolayers indicates that GLN maintains transepithelial resistance and decreases permeability. The mechanisms of these effects remain undefined. We hypothesized that GLN affects proteins involved in the intercellular junctional complex. GLN availability was controlled in Caco-2 monolayers by addition to the medium and treatment with methionine sulfoximine (MSO) to inhibit glutamine synthetase (GS). Expression of TJ proteins, claudin-1, occludin, and zonula occluden (ZO)-1 was measured by immunoblotting. Localization of TJ proteins was evaluated by immunofluorescence light microscopy. Structure of TJ was determined by transmission electron microscopy (TEM). Deprivation of GLN decreased claudin-1, occludin, and ZO-1 protein expression and caused a disappearance of perijunctional claudin-1 and a reduction of occludin but had no effect on ZO-1. TEM revealed that MSO-treated cells in the absence of GLN formed irregular junctional complexes between the apical lateral margins of adjoining cells. These findings indicate that TJ protein expression and cellular localization in Caco-2 cell monolayers rely on GLN. This mechanism may similarly relate to GLN-mediated modulation of intestinal barrier function in stressed animals and humans. PMID- 15130875 TI - Low myo-inositol and high glutamine levels in brain are associated with neuropsychological deterioration after induced hyperammonemia. AB - The neuropsychological effect of hyperammonemia is variable. This study tests the hypothesis that the effect of ammonia on the neuropsychological function in patients with cirrhosis is determined by the ability of the brain to buffer ammonia-induced increase in glutamine within the astrocyte by losing osmolytes like myo-inositol (mI) and not by the magnitude of the induced hyperammonemia. Fourteen cirrhotic patients with no evidence of overt hepatic encephalopathy were given a 75-g amino acid (aa) solution mimicking the hemoglobin molecule to induce hyperammonemia. Measurement of a battery of neuropsychological function tests including immediate memory, ammonia, aa, and short-echo time proton magnetic resonance spectroscopy were performed before and 4 h after administration of the aa solution. Eight patients showed deterioration in the Immediate Memory Test at 4 h. Demographic factors, severity of liver disease, change in plasma ammonia, and aa profiles after the aa solution were similar in those that showed a deterioration compared with those who did not. In patients who showed deterioration in the memory test, the mI-to-creatine ratio (mI/Cr) was significantly lower at baseline than those that did not deteriorate. In contrast, the glutamate/glutamine-to-Cr ratio was significantly greater in the patients that deteriorated. The observation that deterioration in the memory test scores was greater in those with lower mI/Cr supports the hypothesis that the neuropsychological effects of induced hyperammonemia is determined by the capacity of the brain to handle ammonia-induced increase in glutamine. PMID- 15130877 TI - Mice lacking melanin-concentrating hormone receptor 1 demonstrate increased heart rate associated with altered autonomic activity. AB - Melanin-concentrating hormone (MCH) plays an important role in energy balance. The current studies were carried out on a new line of mice lacking the rodent MCH receptor (MCHR1(-/-) mice). These mice confirmed the previously reported lean phenotype characterized by increased energy expenditure and modestly increased caloric intake. Because MCH is expressed in the lateral hypothalamic area, which also has an important role in the regulation of the autonomic nervous system, heart rate and blood pressure were measured by a telemetric method to investigate whether the increased energy expenditure in these mice might be due to altered autonomic nervous system activity. Male MCHR1(-/-) mice demonstrated a significantly increased heart rate [24-h period: wild type 495 +/- 4 vs. MCHR1(-/ ) 561 +/- 8 beats/min (P < 0.001); dark phase: wild type 506 +/- 8 vs. MCHR1(-/-) 582 +/- 9 beats/min (P < 0.001); light phase: wild type 484 +/- 13 vs. MCHR1(-/-) 539 +/- 9 beats/min (P < 0.005)] with no significant difference in mean arterial pressure [wild type 110 +/- 0.3 vs. MCHR1(-/-) 113 +/- 0.4 mmHg (P > 0.05)]. Locomotor activity and core body temperature were higher in the MCHR1(-/-) mice during the dark phase only and thus temporally dissociated from heart rate differences. On fasting, wild-type animals rapidly downregulated body temperature and heart rate. MCHR1(-/-) mice displayed a distinct delay in the onset of this downregulation. To investigate the mechanism underlying these differences, autonomic blockade experiments were carried out. Administration of the adrenergic antagonist metoprolol completely reversed the tachycardia seen in MCHR1(-/-) mice, suggesting an increased sympathetic tone. PMID- 15130876 TI - Limited role for CXC chemokines in the pathogenesis of alpha naphthylisothiocyanate-induced liver injury. AB - Alpha-naphthylisothiocyanate (ANIT) is a hepatotoxin that causes severe neutrophilic inflammation around portal tracts and bile ducts. The chemotactic signals that provoke this inflammatory response are unknown. In this study, we addressed the possibility that ANIT upregulates CXC chemokines in the liver and that these compounds mediate hepatic inflammation and tissue injury after ANIT treatment. Mice treated with a single dose of ANIT (50 mg/kg) exhibited rapid hepatic induction of the CXC chemokine macrophage inflammatory protein-2 (MIP-2). MIP-2 derived primarily from hepatocytes, with no apparent contribution by biliary cells. In ANIT-treated mice, the induction of MIP-2 coincided with an influx of neutrophils to portal zones; this hepatic neutrophil recruitment was suppressed by 50% in mice that lack the receptor for MIP-2 (CXCR2(-/-)). Interestingly, despite their markedly reduced degree of hepatic inflammation, CXCR2(-/-) mice displayed just as much hepatocellular injury and cholestasis after ANIT treatment as wild-type mice. Moreover, after long-term exposure, ANIT CXCR2(-/-) mice developed liver fibrosis that was indistinguishable from that in wild-type mice. In summary, our data show that CXC chemokines are responsible for some of the hepatic inflammation that occurs in response to ANIT but that these compounds are not essential to the pathogenesis of either acute or chronic ANIT hepatotoxicity. PMID- 15130878 TI - Chronic mevastatin modulates receptor-dependent vascular contraction in eNOS deficient mice. AB - We tested the hypothesis that endothelial nitric oxide (NO) synthase (eNOS) derived NO modulates rho-kinase-mediated vascular contraction. Because 3-hydroxy 3-methylglutaryl (HMG)-CoA-reductase inhibition can both upregulate eNOS expression and inhibit rhoA/rho-kinase function, a second hypothesis tested was that statin treatment modulates rho-kinase-mediated contraction and that this can occur independently of eNOS. Contractile responses to the receptor-dependent agonists serotonin and phenylephrine but not to the receptor-independent agent KCl were greater in aortic rings from eNOS-null (eNOS(-/-)) vs. wild-type (eNOS(+/+)) mice. Similarly enhanced responses were seen in eNOS(+/+) rings after acute NOS inhibition. The rho-kinase inhibitor Y-27632 abolished or profoundly attenuated responses to receptor agonists in both eNOS(+/+) and eNOS(-/-) rings, but responses in eNOS(+/+) were more sensitive to Y-27632. Mevastatin treatment (20 mg/kg sc per day, 14 days) reduced responses to serotonin and phenylephrine in female mice of both strains. KCl-induced contractions were slightly smaller in eNOS(+/+)-derived aortic rings only. Levels of plasma cholesterol, and aortic expression of rhoA and rho-kinase, did not differ between groups. Thus eNOS derived NO suppresses rhoA/rho-kinase-mediated vascular contraction. Moreover, a similar suppressive effect on rho-kinase-mediated vasoconstriction by statin therapy occurs independently of effects on eNOS or plasma cholesterol. PMID- 15130879 TI - Amylin receptor blockade stimulates food intake in rats. AB - Amylin is postulated to act as a hormonal signal from the pancreas to the brain to inhibit food intake and regulate energy reserves. Amylin potently reduces food intake, body weight, and adiposity when administered systemically or into the brain. Whether selective blockade of endogenous amylin action increases food intake and adiposity remains to be clearly established. In the present study, the amylin receptor antagonist acetyl-[Asn(30), Tyr(32)] sCT-(8-32) (AC187) was used to assess whether action of endogenous amylin is essential for normal satiation to occur. Non-food-deprived rats received a 3- to 4-h intravenous infusion of AC187 (60-2,000 pmol.kg(-1).min(-1)), either alone or coadministered with a 3-h intravenous infusion of amylin (2.5 or 5 pmol.kg(-1).min(-1)) or a 2-h intragastric infusion of an elemental liquid diet (4 kcal/h). Infusions began just before dark onset. Food intake and meal patterns during the first 4 h of the dark period were determined from continuous computer recordings of changes in food bowl weight. Amylin inhibited food intake by approximately 50%, and AC187 attenuated this response by approximately 50%. AC187 dose-dependently stimulated food intake (maximal increases from 76 to 171%), whether administered alone or with an intragastric infusion of liquid diet. Amylin reduced mean meal size and meal frequency, AC187 attenuated these responses, and AC187 administration alone increased mean meal size and meal frequency. These results support the hypothesis that endogenous amylin plays an essential role in reducing meal size and increasing the postmeal interval of satiety. PMID- 15130880 TI - Systems analysis of digoxin kinetics and inotropic response in the rat heart: effects of calcium and KB-R7943. AB - To gain more insight into the mechanistic processes controlling the kinetics of inotropic response of digoxin in the perfused whole heart, an integrated kinetic model was developed incorporating digoxin uptake, receptor binding (Na(+)-K(+) ATPase inhibition), and cellular events linking receptor occupation and response. The model was applied to data obtained in the single-pass Langendorff-perfused rat heart for external [Ca(2+)] of 0.5 and 1.5 mM under control conditions and in the presence of the reverse-mode Na(+)/Ca(2+) exchange inhibitor KB-R7943 (0.1 microM) in perfusate. Outflow concentration and left ventricular developed pressure data measured for three consecutive doses (15, 30, and 45 microg) in each heart were analyzed simultaneously. While disposition kinetics of digoxin was determined by interaction with a heterogeneous receptor population consisting of a high-affinity/low-capacity and a low-affinity/high- capacity binding site, response generation was >80% mediated by binding to the high-affinity receptor. Digoxin sensitivity increased at lower external [Ca(2+)] due to higher stimulus amplification. Coadministration of KB-R7943 significantly reduced the positive inotropic effect of digoxin at higher doses (30 and 45 microg) and led to a saturated and delayed receptor occupancy-response relationship in the cellular effectuation model. The results provide further evidence for the functional heterogeneity of the Na(+)-K(+)-ATPase and suggest that in the presence of KB R7943 a reduction of the Ca(2+) influx rate via the reverse mode Na(+)/Ca(2+) exchanger might become the limiting factor in digoxin response generation. PMID- 15130881 TI - Differential expression of alpha2D-adrenoceptor and eNOS in aortas from early and later stages of diabetes in Goto-Kakizaki rats. AB - The aim of the present study was to compare vascular dysfunction between the early (12 wk old) and later (36 wk old) stages of spontaneous diabetes in Goto Kakizaki (GK) rats. We also evaluated the aortic expression of the alpha(2D) adrenoceptor and endothelial nitric oxide synthase (eNOS). Vascular reactivity was assessed in thoracic aortas from age-matched control rats and 12- and 36-wk GK rats. Using RT-PCR and immunoblots, we also examined the changes in expression of the alpha(2D-)adrenoceptor and eNOS. In aortas from GK rats (vs. those from age-matched control rats): 1) the relaxation response to ACh was enhanced at 12 wk but decreased at 36 wk; 2) the relaxation response to sodium nitroprusside was decreased at both 12 and 36 wk, 3) norepinephrine (NE)-induced contractility was decreased at 12 wk but not at 36 wk, 4) the expressions of alpha(1B)- and alpha(1D)-adrenoceptors were unaffected, whereas those of alpha(2D)-adrenoceptor and eNOS mRNAs were increased at both 12 and 36 wk; and 5) NE- and ACh-stimulated NO(x) (nitrite and nitrate) levels were increased at 12 wk, although at 36 wk ACh stimulated NO(x) was lower, whereas NE-stimulated NO(x) showed no change. These results clearly demonstrate that enhanced ACh-induced relaxation and impaired NE induced contraction, due to NO overproduction via eNOS and increased alpha(2D) adrenoceptor expression, occur in early-stage GK rats and that the impaired ACh induced relaxation in later-stage GK rats is due to reductions in both NO production and NO responsiveness (but not in eNOS expression). PMID- 15130882 TI - 17beta-estradiol inhibits cyclic strain-induced endothelin-1 gene expression within vascular endothelial cells. AB - It has been well documented previously that 17beta-estradiol (E2) exerts a protective effect on cardiovascular tissue. The possible role of E2 in the regulation of endothelin (ET)-1 production has been previously reported, although the complex mechanisms by which E2 inhibits ET-1 expression are not completely understood. The aims of this study were to examine whether E2 was able to alter strain-induced ET-1 gene expression and also to identify the putative underlying signaling pathways that exist within endothelial cells. For cultured endothelial cells, E2 (1-100 nM), but not 17alpha-estradiol, inhibited the level of strain induced ET-1 gene expression and also peptide secretion. This inhibitory effect elicited by E2 was able to be prevented by the coincubation of endothelial cells with the estrogen receptor antagonist ICI-182,780 (1 microM). E2 also inhibited strain-enhanced NADPH oxidase activity and intracellular reactive oxygen species (ROS) generation as measured by the redox-sensitive fluorescent dye 2',7' dichlorofluorescin diacetate and the level of extracellular signal-regulated kinase (ERK) phosphorylation. Furthermore, the presence of E2 and antioxidants such as N-acetylcysteine and diphenylene iodonium were able to elicit a decrease in the level of strain-induced ET-1 secretion, ET-1 promoter activity, ET-1 mRNA, ERK phosphorylation, and activator protein-1 binding activity. In summary, we demonstrated, for the first time, that E2 inhibits strain-induced ET-1 gene expression, partially by interfering with the ERK pathway via the attenuation of strain-induced ROS generation. Thus this study delivers important new insight regarding the molecular pathways that may contribute to the proposed beneficial effects of estrogen on the cardiovascular system. PMID- 15130883 TI - Myocardial insulin resistance associated with chronic hypertriglyceridemia and increased FFA levels in Type 2 diabetic patients. AB - We evaluated the influence of chronic hypertriglyceridemia and endothelial dysfunction on myocardial glucose uptake (MGU) in Type 2 diabetic patients without coronary heart disease. Patients were divided into two groups according to fasting triglyceride (TG) levels: 5.4 +/- 1.1 and 1.5 +/- 0.3 mmol/l for high- and normal-TG groups, respectively. Five subjects were assigned to the high-TG group and 11 to the normal-TG group. Age, gender, body mass index, systolic and diastolic blood pressure, glucose, insulin, HbA1c, cholesterol, and HDL cholesterol were similar in the two groups, whereas free fatty acid (FFA) levels were higher in the high-TG group basally and at the end of the clamp. Furthermore, five healthy subjects were subjected to the same protocol and used as the control group. MGU was assessed by using 18F-labeled 2-fluoro-2-deoxy-D glucose under hyperglycemic-hyperinsulinemic conditions. Basal endothelin-1 and nitric oxide levels were significantly higher in the high-TG group than in the normal-TG and control groups, and cGMP and maximal postischemic vasodilation were significantly decreased in the high-TG group compared with the normal-TG and control groups. However, significant alterations were found in the same parameters in the normal-TG group compared with the control group. By the end of the hyperglycemic clamp, in the high-TG group, MGU was approximately 40 and 65% of that in the normal-TG and control groups. MGU negatively correlated with TG, FFA, and endothelin-1, whereas a positive correlation was found with cGMP and maximal postischemic vasodilation. In conclusion, increased TG and FFA levels are risks, in addition to Type 2 diabetes mellitus, for myocardial insulin resistance, endothelial dysfunction, and alteration of nitric oxide/cGMP levels. PMID- 15130884 TI - Transfection of CYP4A1 cDNA decreases diameter and increases responsiveness of gracilis muscle arterioles to constrictor stimuli. AB - Cytochrome P-450-4A1 (CYP4A1) is an omega-hydroxylase that catalyzes the metabolism of arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE). The goal of this study was to determine the vasomotor consequences of vascular overexpression of CYP4A1. Isolated rat gracilis muscle arterioles transfected ex vivo with an expression plasmid containing CYP4A1 cDNA expressed more CYP4A protein than vessels transfected with the control plasmid. In arterioles pressurized to 80 mmHg, the internal diameter of vessels transfected with CYP4A1 cDNA (55 +/- 3 microm) was surpassed (P < 0.05) by that of vessels transfected with control plasmid (97 +/- 4 microm). Treatment with a CYP4A inhibitor (N methylsulfonyl-12,12-dibromododec-11-enamide; DDMS) or with an antagonist of 20 HETE actions [20-hydroxyeicosa-6(Z),15(Z)-dienoic acid; 20-HEDE] elicited robust dilation of arterioles transfected with CYP4A1 cDNA, whereas the treatment had little or no effect in vessels transfected with control plasmid. Examination of the intraluminal pressure-internal diameter relationship revealed that pressure increments over the range of 40-100 mmHg elicited a more intense (P < 0.05) myogenic constrictor response in arterioles transfected with CYP4A1 cDNA than in those with control plasmid. Arterioles transfected with CYP4A1 cDNA also displayed enhanced sensitivity to the constrictor action of phenylephrine. Treatment with DDMS or 20-HEDE greatly attenuated the constrictor responsiveness to both constrictor stimuli in vessels overexpressing CYP4A1, whereas the treatment had much less effect in control vessels. These data suggest that CYP4A1 overexpression promotes constriction of gracilis muscle arterioles by intensifying the responsiveness of vascular smooth muscle to constrictor stimuli. This effect of CYP4A1 overexpression appears to be mediated by a CYP4A1 product. PMID- 15130887 TI - Effect of shear stress on microvessel network formation of endothelial cells with in vitro three-dimensional model. AB - Shear stress stimulus is expected to enhance angiogenesis, the formation of microvessels. We determined the effect of shear stress stimulus on three dimensional microvessel formation in vitro. Bovine pulmonary microvascular endothelial cells were seeded onto collagen gels with basic fibroblast growth factor to make a microvessel formation model. We observed this model in detail using phase-contrast microscopy, confocal laser scanning microscopy, and electron microscopy. The results show that cells invaded the collagen gel and reconstructed the tubular structures, containing a clearly defined lumen consisting of multiple cells. The model was placed in a parallel-plate flow chamber. A laminar shear stress of 0.3 Pa was applied to the surfaces of the cells for 48 h. Promotion of microvessel network formation was detectable after approximately 10 h in the flow chamber. After 48 h, the length of networks exposed to shear stress was 6.17 (+/-0.59) times longer than at the initial state, whereas the length of networks not exposed to shear stress was only 3.30 (+/-0.41) times longer. The number of bifurcations and endpoints increased for networks exposed to shear stress, whereas the number of bifurcations alone increased for networks not exposed to shear stress. These results demonstrate that shear stress applied to the surfaces of endothelial cells on collagen gel promotes the growth of microvessel network formation in the gel and expands the network because of repeated bifurcation and elongation. PMID- 15130885 TI - Correction of motion artifact in transmembrane voltage-sensitive fluorescent dye emission in hearts. AB - Fast voltage-sensitive dyes are widely used to image cardiac electrical activity. Typically, the emission spectrum of these fluorochromes is wavelength shifted with altered membrane potential, but the optical signals obtained also decay with time and are affected by contraction. Ratiometry reduces, but may not fully remove, these artifacts. An alternate approach has been developed in which the time decay in simultaneously acquired short- and long-wavelength signals is characterized nonparametrically and removed. Motion artifact is then identified as the time-varying signal component common to both decay-corrected signals and subtracted. Performance of this subtraction technique was compared with ratiometry for intramural optical signals acquired with a fiber-optic probe in an isolated, Langendorff-perfused pig heart preparation (n = 4) stained with di-4 ANEPPS. Perfusate concentration of 2,3-butanedione monoxime was adjusted (7.5 12.5 mM) to alter contractile activity. Short-wavelength (520-600 nm) and long wavelength (>600 nm) signals were recorded over 8-16 cardiac cycles at 6 sites across the left ventricular free wall in sinus rhythm and during pacing. A total of 451 such data sets were acquired. Appreciable wall motion was observed in 225 cases, with motion artifact classed as moderate (less than modulation due to action potential) in 187 and substantial (more than modulation due to action potential) in 38. In all cases, subtraction performed as well as, or better than, ratiometry in removing motion artifact and decay. Action potential morphology was recovered more faithfully by subtraction than by ratiometry in 58 of 187 and 31 of 38 cases with moderate and substantial motion artifact, respectively. This novel subtraction approach may therefore provide a means of reducing the concentration of uncoupling agents used in cardiac optical mapping studies. PMID- 15130886 TI - Inotropic effects of ETB receptor stimulation and their modulation by endocardial endothelium, NO, and prostaglandins. AB - Endothelin (ET)-1 acts on ETA and ETB receptors. The latter include ETB1 (endothelial) and ETB2 (muscular) subtypes, which mediate opposite effects on vascular tone. This study investigated, in rabbit papillary muscles (n = 84), the myocardial effects of ETB stimulation. ET-1 (10(-9) M) was given in the absence or presence of BQ-123 (ETA antagonist). The effects of IRL-1620 (ETB1 agonist, 10(-10)-10(-6) M) or sarafotoxin S6c (ETB agonist, 10(-10)-10(-6) M) were evaluated in muscles with intact or damaged endocardial endothelium (EE); intact EE, in the presence of NG-nitro-L-arginine (L-NNA); and intact EE, in the presence of indomethacin (Indo). Sarafotoxin S6c effects were also studied in the presence of BQ-788 (ETB2 antagonist). ET-1 alone increased 64 +/- 18% active tension (AT) but decreased it by 4 +/- 2% in the presence of BQ-123. In muscles with intact EE, sarafotoxin S6c alone did not significantly alter myocardial performance. Sarafotoxin S6c (10(-6) M) increased, however, AT by 120 +/- 27% when EE was damaged and by 39 +/- 8% or 23 +/- 6% in the presence of l-NNA or Indo, respectively. In the presence of BQ-788, sarafotoxin S6c decreased AT (21 +/- 3% at 10(-6) M) in muscles with intact EE, an effect that was abolished when EE was damaged. IRL-1620 also decreased AT (22 +/- 3% at 10(-6) M) in muscles with intact EE, an effect that was abolished when EE was damaged or in the presence of L-NNA or Indo. In conclusion, the ETB-mediated negative inotropic effect is presumably due to ETB1 stimulation, requires an intact EE, and is mediated by NO and prostaglandins, whereas the ETB-mediated positive inotropic effect, observed when EE was damaged or NO and prostaglandins synthesis inhibited, is presumably due to ETB2 stimulation. PMID- 15130888 TI - Vascular transfer of adenovirus is augmented by nitric oxide in the rat heart. AB - Reversible opening of the endothelial barrier remains a major obstacle when hearts are transfected via the coronary system. Our aim was to establish an experimental system permitting the continuous analysis of vascular transfer of virus in the intact heart. Isolated saline-perfused rat hearts were inverted and covered with a latex cap to collect interstitial transudate (IT) on the pericardial surface. Adenovirus (10(9) pfu/ml) was stably labeled with rhodamine fluorescent dye. Analysis of IT and coronary perfusate revealed that under baseline conditions, adenovirus in the IT reached 75% of its vascular concentration within 3 min. The nitric oxide-donors S-nitroso-N-acetyl penicillamine (SNAP) and bradykinin (BK) were the most effective substances to increase total IT volume and adenoviral interstitial concentration. Perfusion with 9% serum markedly reduced IT volume flow and delayed the SNAP/BK effect. Our findings demonstrate that SNAP and BK effectively increased coronary transfer of adenovirus suggesting that the inverted isolated heart is a suitable model to optimize vascular transfer of virus under standardized conditions. PMID- 15130889 TI - Increases in CSF [Na+] precede the increases in blood pressure in Dahl S rats and SHR on a high-salt diet. AB - In Dahl salt-sensitive (S) and salt-resistant (R) rats, and spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats, at 5-6 wk of age, a cannula was placed in the cisterna magna, and cerebrospinal fluid (CSF) was withdrawn continuously at 75 microl/12 h. CSF was collected as day- and nighttime samples from rats on a regular salt intake (0.6% Na+; R-Na) and then on a high salt intake (8% Na+; H-Na). In separate groups of rats, the abdominal aorta was cannulated and blood pressure (BP) and heart rate (HR) measured at 10 AM and 10 PM, with rats first on R-Na and then on H-Na. On H-Na, CSF [Na+] started to increase in the daytime of day 2 in Dahl S rats and of day 3 in SHR. BP and HR did not rise until day 3 in Dahl S rats and day 4 in SHR. In Dahl R and WKY rats, high salt did not change CSF [Na+], BP, or HR. In a third set of Dahl S rats, sampling of both CSF and BP was performed in each individual rat. Again, significant increases in CSF [Na+] were observed 1-2 days earlier than the increases in BP and HR. In a fourth set of Dahl S rats, BP and HR were recorded continuously by means of radiotelemetry for 5 days on R-Na and 8 days on H-Na. On H-Na, BP (but not HR) increased first in the nighttime of day 2. In another set of Dahl S rats, intracerebroventricular infusion of antibody Fab fragments binding ouabain-like compounds (OLC) with high affinity prevented the increase in BP and HR by H-Na but further increased CSF [Na+]. Finally, in Wistar rats on H Na, intracerebroventricular infusion of ouabain increased BP and HR but decreased CSF [Na+]. Thus, in both Dahl S and SHR on H-Na, increases in CSF [Na+] preceded the increases in BP and HR, consistent with a primary role of increased CSF [Na+] in the salt-induced hypertension. An increase in brain OLC in response to the initial increase in CSF [Na+] appears to attenuate further increases in CSF [Na+] but at the "expense" of sympathoexcitation and hypertension. PMID- 15130890 TI - A constitutive formulation of arterial mechanics including vascular smooth muscle tone. AB - A pseudo-strain energy function (pseudo-SEF) describing the biomechanical properties of large conduit arteries under the influence of vascular smooth muscle (VSM) tone is proposed. In contrast to previous models that include the effects of smooth muscle contraction through generation of an active stress, in this study we consider the vascular muscle as a structural element whose contribution to load bearing is modulated by the contraction. This novel pseudo SEF models not only arterial mechanics at maximal VSM contraction but also the myogenic contraction of the VSM in response to local increases in stretch. The proposed pseudo-SEF was verified with experimentally obtained pressure-radius curves and zero-stress state configurations from rat carotid arteries displaying distinct differences in VSM tone: arteries from normotensive rats displaying minimal VSM tone and arteries from hypertensive rats exhibiting significant VSM tone. The pressure-radius curves were measured in three different VSM states: fully relaxed, maximally contracted, and normal VSM tone. The model fitted the experimental data very well (r2 > 0.99) in both the normo- and hypertensive groups for all three states of VSM activation. The pseudo-SEF was used to illustrate the localized reduction of circumferential stress in the arterial wall due to normal VSM tone, suggesting that the proposed pseudo-SEF can be of general utility for describing stress distribution not only under passive VSM conditions, as most SEFs proposed so far, but also under physiological and pathological conditions with varying levels of VSM tone. PMID- 15130891 TI - A beneficial role of cardiac P2X4 receptors in heart failure: rescue of the calsequestrin overexpression model of cardiomyopathy. AB - The P2X4 purinergic receptor (P2X4R) is a ligand-gated ion channel. Its activation by extracellular ATP results in Ca2+ influx. Transgenic cardiac overexpression of the human P2X4 receptor showed an in vitro phenotype of enhanced basal contractility. The objective here was to determine the in vivo cardiac physiological role of this receptor. Specifically, we tested the hypothesis that this receptor plays an important role in modulating heart failure progression. Transgenic cardiac overexpression of canine calsequestrin (CSQ) showed hypertrophy, heart failure, and premature death. Crossing the P2X4R mouse with the CSQ mouse more than doubled the lifespan (182 +/- 91 days for the binary CSQ/P2X4R mouse, n = 35) of the CSQ mouse (71.3 +/- 25.4 days, n = 50, P < 0.0001). The prolonged survival in the binary CSQ/P2X4R mouse was associated with an improved left ventricular weight-to-body weight ratio and a restored beta adrenergic responsiveness. The beneficial phenotype of the binary mouse was not associated with any downregulation of the CSQ level but correlated with improved left ventricular developed pressure and +/-dP/dt. The enhanced cardiac performance was manifested in young binary animals and persisted in older animals. The increased contractility likely underlies the survival benefit from P2X4 receptor overexpression. An increased expression or activation of this receptor may represent a new approach in the therapy of heart failure. PMID- 15130892 TI - Diabetes-related changes in cAMP-dependent protein kinase activity and decrease in relaxation response in rat mesenteric artery. AB - Using superior mesenteric artery rings isolated from age-matched controls and streptozotocin (STZ)-induced diabetic rats, we recently demonstrated that EDHF type relaxation is impaired in STZ-induced diabetic rats, possibly due to a reduced action of cAMP via increased phosphodiesterase (PDE) activity (Matsumoto T, Kobayashi T, and Kamata K. Am J Physiol Heart Circ Physiol 285: H283-H291, 2003). Here, we investigated the activity and expression of cAMP-dependent protein kinase (PKA), an enzyme that is produced by a pleiotropic and plays key roles in the transduction of many external signals through the cAMP second messenger pathway and in cAMP-mediated vasorelaxation. The relaxation induced by cilostamide, a selective PDE3 inhibitor, was significantly weaker in superior mesenteric artery rings from STZ-induced diabetic rats than in those from age matched controls. The relaxation responses to 8-bromo-cAMP (8Br-cAMP) and N6,O2 dibutyryl-adenosine-cAMP (db-cAMP), a cell-permeant cAMP analog, were also impaired in the STZ diabetic group. PKA activity in the db-cAMP-treated mesenteric artery was significantly lower in the STZ diabetic group. The expression levels of the mRNA and protein for PKA catalytic subunit Cat-alpha were significantly decreased in the STZ diabetic group, but those for PKA regulatory subunit isoform RII-beta were increased. We conclude that the abnormal vascular relaxation responsiveness seen in STZ-induced diabetic rats may be attributable not only to increased PDE activity but also to decreased PKA activity. Possibly, the decreased PKA activity may result from an imbalance between PKA catalytic and regulatory subunit expressions. PMID- 15130893 TI - Intracellular signaling pathways involved in Gas6-Axl-mediated survival of endothelial cells. AB - Gas6 is a gamma-carboxylated ligand for the receptor tyrosine kinase Axl. Gas6 Axl interactions can rescue endothelial cells from apoptosis, and this study examined the intracellular signaling mechanisms responsible for this phenomenon. Using flow cytometry, we first confirmed that Gas6 can abrogate apoptosis induced by serum starvation of primary cultures of human umbilical vein endothelial cells (HUVECs). This effect is mediated through phosphorylation of the serine-threonine kinase Akt, with maximal phosphorylation observed after 4 h of treatment with 100 ng/ml Gas6. Inhibition of Akt phosphorylation and abrogation of gas6-mediated survival of HUVECs by wortmannin implicated phosphatidylinositol 3-kinase as the mediator of Akt phosphorylation. Dominant negative Akt constructs largely abrogated the protective effect of Gas6 on HUVECs, underscoring the importance of Akt activation in Gas6-mediated survival. Several downstream regulators of this survival pathway were identified in HUVECs, namely, NF-kappaB as well as the antiapoptotic and proapoptotic proteins Bcl-2 and caspase 3, respectively. We showed that NF-kappaB is phosphorylated early after Gas6 treatment as evidenced by doublet formation on Western blotting. As well, the level of Bcl-2 protein increased, supporting the notion that the Bcl-2 antiapoptotic pathway is stimulated. The levels of expression of the caspase 3 activation products p12 and p20 decreased with Gas6 treatment, consistent with a reduction in proapoptotic caspase 3 activation. Taken together, these experiments provide new information about the mechanism underlying Gas6 protection from apoptosis in primary endothelial cell cultures. PMID- 15130895 TI - Regulation of calcium signaling by polycystin-2. AB - Autosomal dominant PKD (ADPKD) is a common lethal genetic disorder characterized by progressive development of fluid-filled cysts in the kidney and other target organs. ADPKD is caused by mutations in the PKD1 and PKD2 genes, encoding the transmembrane proteins polycystin-1 (PC1) and polycystin-2 (PC2), respectively. Although the function and putative interacting ligands of PC1 are largely unknown, recent evidence indicates that PC2 behaves as a TRP-type Ca(2+) permeable nonselective cation channel. The PC2 channel is implicated in the transient increase in cytosolic Ca(2+) in renal epithelial cells and may be linked to the activation of subsequent signaling pathways. Recent studies also indicate that PC1 functionally interacts with PC2 such that the PC1-PC2 channel complex is an obligatory novel signaling pathway implicated in the transduction of environmental signals into cellular events. The present review purposely avoids issues of regulation of PC2 expression and trafficking and focuses instead on the evidence for the TRP-type cation channel function of PC2. How its role as a cation channel may unmask mechanisms that trigger Ca(2+) transport and regulation is the focus of attention. PC2 channel function may be essential in renal cell function and kidney development. Nonrenal-targeted expression of PC2 and related proteins, including the cardiovascular system, also suggests previously unforeseeable roles in signal transduction. PMID- 15130894 TI - Role of the calcium-sensing receptor in parathyroid gland physiology. AB - The calcium-sensing receptor (CaSR) represents the molecular mechanism by which parathyroid cells detect changes in blood ionized calcium concentration and modulate parathyroid hormone (PTH) secretion to maintain serum calcium levels within a narrow physiological range. Much has been learned in recent years about the diversity of signal transduction through the CaSR and the various factors that affect receptor expression. Beyond its classic role as a determinant of calcium-regulated PTH secretion, signaling through the CaSR also influences both gene transcription and cell proliferation in parathyroid cells. The CaSR thus serves a broad physiological role by integrating several distinct aspects of parathyroid gland function. The current review summarizes recent developments that enhance our understanding of the CaSR and its fundamental importance in parathyroid gland physiology. PMID- 15130897 TI - Effect of aldosterone on renal transforming growth factor-beta. AB - Aldosterone participates in the pathophysiology of several models of progressive chronic renal disease. Because of the causal connection between transforming growth factor-beta(1) (TGF-beta) and scarring in many such models, we hypothesized that aldosterone could evoke TGF-beta in the kidney. Aldosterone infusion for 3 days in otherwise normal rats caused a more than twofold increase in TGF-beta excretion without changes in systolic pressure or evidence of kidney damage. Concurrent treatment with amiloride did not alter this effect, indicating that aldosterone's stimulation of TGF-beta was independent of its regulation of sodium or potassium transport. However, concurrent treatment with spironolactone did block the increase in TGF-beta, indicating that the effect depends on the mineralocorticoid receptor. Renal mRNA for serum glucocorticoid kinase rose, but no change in TGF-beta message occurred, suggesting posttranscriptional enhancement of renal TGF-beta. In summary, aldosterone provokes renal TGF-beta, and this action may contribute to aldosterone's fibrotic propensity. PMID- 15130896 TI - Inhibition of erythrocyte phosphatidylserine exposure by urea and Cl-. AB - Osmotic shock by addition of sucrose to the medium stimulates erythrocyte sphingomyelinase with subsequent ceramide formation and triggers Ca(2+) entry through stimulation of cation channels. Both ceramide and Ca(2+) activate an erythrocyte scramblase, leading to breakdown of phosphatidylserine asymmetry, a typical feature of apoptosis. Because erythrocytes are regularly exposed to osmotic shock during passage of kidney medulla, the present study explored the influence of NaCl and urea on erythrocyte phosphatidylserine exposure as determined by annexin binding. The percentage of annexin-binding erythrocytes increased from <5 to 80 +/- 4% (n = 4) upon addition of 650 mM sucrose, an effect paralleled by activation of the cation channel and stimulation of ceramide formation. The number of annexin-binding erythrocytes increased only to 18% after addition of 325 mM NaCl and was not increased by addition of 650 mM urea. According to whole cell patch-clamp experiments, the cation conductance was activated by replacement of extracellular Cl(-) with gluconate at isotonic conditions or by addition of hypertonic sucrose or urea. Although stimulating the cation conductance, urea abrogated the annexin binding and concomitant increase of ceramide levels induced by osmotic cell shrinkage. In vitro sphingomyelinase assays demonstrated a direct inhibitory effect of urea on sphingomyelinase activity. Urea did not significantly interfere with annexin binding after addition of ceramide. In conclusion, both Cl(-) and urea blunt erythrocyte phosphatidylserine exposure after osmotic shock. Whereas Cl(-) is effective through inhibition of the cation conductance, urea exerts its effect through inhibition of sphingomyelinase, thus blunting formation of ceramide. PMID- 15130898 TI - PKC expression is regulated by dietary K intake and mediates internalization of SK channels in the CCD. AB - We have used Western blot analysis and immunocytochemistry to determine the effect of dietary K intake on the expression of protein kinase C (PKC) isoforms in the kidney. Western blot has demonstrated that conventional PKC isoforms (alpha and beta), novel PKC isoforms (delta, epsilon, and eta), and atypical PKC isoforms (zeta) are expressed in the renal cortex and outer medulla. Moreover, a low K intake significantly increases the expression of PKC-epsilon in the renal cortex and outer medulla but does not change the expression of PKC-alpha, PKC beta, PKC-delta, PKC-eta, and PKC-zeta. Also, immunocytochemistry shows that PKC epsilon isoform is expressed in the cortical collecting duct (CCD) and outer medullary collecting duct (OMCD) and that the intensity of PKC-epsilon staining is higher in the kidney from rats on a K-deficient diet than those on a control diet. Also, we used the patch-clamp technique to study the role of PKC in mediating internalization of ROMK (Kir 1.1)-like small-conductance K (SK) channels induced by phenylarsine oxide (PAO), an agent that inhibits protein tyrosine phosphatase and has been shown to stimulate the internalization of the SK channel in the CCD (Sterling H, Lin DH, Qu RM, Dong K, Herbert SC, and Wang WH. J Biol Chem 277: 4317-4323, 2002). Inhibition of PKC with calphostin C and GF 109203x had no significant effect on channel activity but abolished the inhibitory effect of PAO on SK channels. In conclusion, a low K intake increases the expression of PKC-epsilon isoform in the renal cortex and outer medulla, and PKC is involved in mediating the internalization of SK channels in the CCD induced by stimulation of protein tyrosine kinase activity. PMID- 15130900 TI - Urinary bladder cancer in Wegener's granulomatosis: risks and relation to cyclophosphamide. AB - OBJECTIVE: To assess and characterise the risk of bladder cancer, and its relation to cyclophosphamide, in patients with Wegener's granulomatosis. METHODS: In the population based, nationwide Swedish Inpatient Register a cohort of 1065 patients with Wegener's granulomatosis, 1969-95, was identified. Through linkage with the Swedish Cancer Register, all subjects in this cohort diagnosed with bladder cancer were identified. Nested within the cohort, a matched case-control study was performed to estimate the association between cyclophosphamide and bladder cancer using odds ratios (ORs) as relative risk. In the cohort the cumulative risk of bladder cancer after Wegener's granulomatosis, and the relative prevalence of a history of bladder cancer at the time of diagnosis of Wegener's granulomatosis, were also estimated. RESULTS: The median cumulative doses of cyclophosphamide among cases (n = 11) and controls (n = 25) were 113 g and 25 g, respectively. The risk of bladder cancer doubled for every 10 g increment in cyclophosphamide (OR = 2.0, 95% confidence interval (CI) 0.8 to 4.9). Treatment duration longer than 1 year was associated with an eightfold increased risk (OR = 7.7, 95% CI 0.9 to 69). The absolute risk for bladder cancer in the cohort reached 10% 16 years after diagnosis of Wegener's granulomatosis, and a history of bladder cancer was (non-significantly) twice as common as expected at the time of diagnosis of Wegener's granulomatosis. CONCLUSION: The results indicate a dose-response relationship between cyclophosphamide and the risk of bladder cancer, high cumulative risks in the entire cohort, and also the possibility of risk factors operating even before Wegener's granulomatosis. PMID- 15130899 TI - A good response to early DMARD treatment of patients with rheumatoid arthritis in the first year predicts remission during follow up. AB - OBJECTIVE: To describe the frequency and duration of remission in the Utrecht rheumatoid arthritis cohort of patients followed since diagnosis, and the clinical and treatment characteristics of patients with remission v those without. METHODS: In 1990 the Utrecht rheumatoid arthritis cohort study group started a clinical trial in which patients with recent onset of rheumatoid arthritis (<1 year) were randomised into four treatment groups: hydroxychloroquine (n = 169); intramuscular gold (n = 163); methotrexate (n = 166); and pyramid (n = 64). After two years, rheumatologists were allowed to prescribe any disease modifying antirheumatic drug. Remission was defined as: duration of morning stiffness < or =15 min, mean VAS pain < or =10 mm, Thompson joint score < or =10, and ESR < or =30 mm/h during at least six months. Cox regression analysis was used to determine baseline clinical, demographic, and treatment predictors of remission. RESULTS: Mean follow up duration was 62 months. Thirty six per cent achieved at least one period of remission. Median duration between diagnosis and the first remission period was 15 months for the intramuscular gold group, 18 months for the methotrexate and hydroxychloroquine groups, and 24 months for the pyramid group (NS). Predictors of remission were early response to initial treatment, less pain, rheumatoid factor negativity, and lower joint score at baseline. CONCLUSIONS: After a mean follow up duration of 62 months, only 36% of the patients had fulfilled the remission criteria at least once. A good response to treatment during the first year seems to be independently associated with remission rather than initial treatment alone. PMID- 15130901 TI - Pain and joint mobility explain individual subdimensions of the health assessment questionnaire (HAQ) disability index in patients with rheumatoid arthritis. AB - OBJECTIVE: To explore the associations between individual subdimensions of the health assessment questionnaire (HAQ) and clinical variables in patients with rheumatoid arthritis. METHODS: 304 patients with rheumatoid arthritis (73% female, mean (SD) age, 58 (13) years; disease duration 6 (9) years, 69% rheumatoid factor positive) completed the HAQ for functional capacity (0-3) and a 100 mm visual analogue scale for pain. Grip strength, range of motion of the large joints, Larsen score for radiographic damage of hand and foot joints, and the number of tender and swollen joints were recorded. A logit regression model was used to study associations between subdimensions of the HAQ and other variables. RESULTS: Mean (range) total HAQ score was 0.92 (0 to 2.88) and varied from 0.73 to 1.04 in the subdimensions. Disability was lowest in the "walking" and highest in the "reach" subdimension. Pain was an explanatory variable in all individual subdimensions. Decreased grip strength, limitation of shoulder and wrist motion, and a larger number of swollen and tender joints in the upper extremities were related to several subdimensions. A higher pain score and swollen joint count in the upper extremities, decreased grip strength, and limited motion of wrist, shoulder, and knee joints explained increased disability (higher total HAQ scores). CONCLUSIONS: In patients with rheumatoid arthritis, pain and range of movements of joints have the greatest impact on individual subdimensions of the HAQ. Extent of radiographic damage in peripheral joints and the number of swollen and tender joints are of lesser importance for function. PMID- 15130902 TI - Evaluation of clinically relevant states in patient reported outcomes in knee and hip osteoarthritis: the patient acceptable symptom state. AB - BACKGROUND: The patient acceptable symptom state (PASS) is the value beyond which patients can consider themselves well. This concept can help in interpreting results of clinical trials. OBJECTIVE: To determine the PASS estimate for patients with knee and hip osteoarthritis (OA) by assessing pain, patient's global assessment of disease activity, and functional impairment. METHODS: A 4 week prospective multicentre cohort study of 1362 outpatients with knee or hip OA was carried out. Data on assessment of pain and patient's global assessment of disease, measured on visual analogue scales, and functional impairment, measured on the Western Ontario McMaster Universities Osteoarthritis Index (WOMAC) function subscale, were collected at baseline and final visits. The patients assessed their satisfaction with their current state at the final visit. An anchoring method based on the patient's opinion was used. RESULTS: For patients with knee and hip OA, the estimates of PASS were, respectively, 32.3 and 35.0 mm for pain, 32.0 and 34.6 mm for patient global assessment of disease activity, and 31.0 and 34.4 points for WOMAC function score. The PASS varied moderately across the tertiles of baseline scores but not across age, disease duration, or sex. CONCLUSION: The use of PASS in clinical trials would provide more meaningful results expressed as a proportion of patients in an acceptable symptom state. PMID- 15130903 TI - Proteome comparison of alveolar macrophages with monocytes reveals distinct protein characteristics. AB - Alveolar macrophages (AMs) are a subset of tissue macrophages situated in the alveolar milieu. Compared with their precursor blood monocytes, AMs exhibit distinct physiologic functions unique to their anatomic location. However, the molecular details that control monocyte differentiation into AMs remain unknown. This study employed a proteomic approach to define protein characteristics that distinguish AMs from monocytes. AMs and monocytes were obtained from six nonsmoking, healthy donors. Whole cell lysates from each donor's AMs and monocytes were analyzed by two-dimensional (2D) gel electrophoreses. The protein density for each protein spot in a 2D gel was compared between these two cell types. Proteins that demonstrated consistent level changes of greater than 2.5 fold in all six donors were subjected to tandem mass spectrometry for protein identity. Using this process, we revealed proteome changes in AMs that relate to their physiologic roles in proteolysis, actin reorganization, and cellular adaptation in the unique alveolar milieu. By comparison, blood monocytes displayed higher levels of the proteins involved in transcription, metabolism, inflammation, and in the control of proteolysis. These results provide new insights into the biology of mononuclear phagocytes and set a basis for future causality studies. PMID- 15130905 TI - Bronchiectasis, exacerbation indices, and inflammation in chronic obstructive pulmonary disease. AB - Relationships between high-resolution computed tomography (HRCT) findings in chronic obstructive pulmonary disease (COPD) and bacterial colonization, airway inflammation, or exacerbation indices are unknown. Fifty-four patients with COPD (mean [SD]: age, 69 [7] years; FEV(1), 0.96 [0.33] L; FEV(1) [percent predicted], 38.1 [13.9]%; FEV(1)/forced vital capacity [percent predicted], 40.9 [11.8]%; arterial partial pressure of oxygen, 8.77 [1.11] kPa; history of smoking, 50.5 [33.5] smoking pack-years) underwent HRCT scans of the chest to quantify the presence and extent of bronchiectasis or emphysema. Exacerbation indices were determined from diary cards over 2 years. Quantitative sputum bacteriology and cytokine measurements were performed. Twenty-seven of 54 patients (50%) had bronchiectasis on HRCT, most frequently in the lower lobes (18 of 54, 33.3%). Patients with bronchiectasis had higher levels of airway inflammatory cytokines (p = 0.001). Lower lobe bronchiectasis was associated with lower airway bacterial colonization (p = 0.004), higher sputum interleukin-8 levels (p = 0.001), and longer symptom recovery time at exacerbation (p = 0.001). No relationship was seen between exacerbation frequency and HRCT changes. Evidence of moderate lower lobe bronchiectasis on HRCT is common in COPD and is associated with more severe COPD exacerbations, lower airway bacterial colonization, and increased sputum inflammatory markers. PMID- 15130904 TI - Respiratory syncytial virus in allergic lung inflammation increases Muc5ac and gob-5. AB - Respiratory syncytial virus (RSV) is associated with wheezing and childhood asthma. We previously reported that RSV infection prolongs methacholine-induced airway hyperresponsiveness in ovalbumin (OVA)-sensitized mice. In addition, allergically sensitized RSV-infected (OVA/RSV) mice had more abundant airway epithelial mucus production compared with OVA mice 14 days after infection, whereas there was almost no mucus in mice that were only RSV infected. We hypothesized that this increased mucus was associated with mucosal expression of Muc5ac, a mucus gene expression in airways, and gob-5, a member of the Ca(2)(+) activated chloride channel family. By histochemical analysis, we found that there was significantly increased staining for gob-5 and Muc5ac in the airways of OVA/RSV mice compared with either OVA mice or allergically sensitized mice that were challenged with inactivated RSV, and virtually no detectable staining in the RSV group. These findings were confirmed by Western blot analysis. The increased mucus expression in the OVA/RSV group was associated with increased lung levels of interleukin-17, a factor known to stimulate airway mucin gene expression. The impact of virus infection combined with allergic inflammation on mucus production may partially explain the more severe disease and airway hyperresponsiveness associated with RSV in the setting of atopy. PMID- 15130906 TI - Noninvasive assessment of airway alterations in smokers: the small airways revisited. AB - It has been shown that structural changes in small airways of smokers with average smoking histories greater than 35 pack-years could be reflected in the single-breath washout test. The more sophisticated multiple breath washout test (MBW) has the potential to anatomically locate the affected small airways in acinar and conductive lung zones through increased phase III slope indices S(acin) and S(cond), respectively. Pulmonary function, S(acin), and S(cond) were obtained in 63 normal never-smokers and in 169 smokers classified according to smoking history (< 10 pack-years; 10-20 pack-years; 20-30 pack-years; > 30 pack years). Compared with never-smokers, significant changes in S(acin) (p = 0.02), S(cond) (p < 0.001), and diffusing capacity (DL(CO); p < 0.001) were detected from greater than 10 pack-years onwards. Spirometric abnormality was significant only from greater than 20 pack-years onwards. In smokers with greater than 30 pack-years and DL(CO) less than 60% predicted, the presence of emphysema resulted in disproportionally larger S(acin) than S(cond) increases. We conclude that S(cond) and S(acin) can noninvasively detect airway changes from as early as 10 pack-years onwards, locating the earliest manifestations of smoking-induced small airways alterations around the acinar entrance. In these early stages, the associated DL(CO) decrease may be a reflection of ventilation heterogeneity rather than true parenchymal destruction. In more advanced stages of smoking induced lung disease, differential patterns of S(acin) and S(cond) are characteristic of the presence of parenchymal destruction in addition to peripheral airways alterations. PMID- 15130907 TI - T cell-based tracking of multidrug resistant tuberculosis infection after brief exposure. AB - Molecular epidemiology indicates significant transmission of Mycobacterium tuberculosis after casual contact with infectious tuberculosis cases. We investigated M. tuberculosis transmission after brief exposure using a T cell based assay, the enzyme-linked-immunospot (ELISPOT) for IFN-gamma. After childbirth, a mother was diagnosed with sputum smear-positive multidrug-resistant tuberculosis. Forty-one neonates and 47 adults were present during her admission on the maternity unit; 11 weeks later, all underwent tuberculin skin testing (TST) and ELISPOT. We correlated test results with markers of exposure to the index case. The participants, who were asymptomatic and predominantly had no prior tuberculosis exposure, had 6.05 hours mean exposure (range: 0-65 hours) to the index case. Seventeen individuals, including two newborns, were ELISPOT positive, and ELISPOT results correlated significantly with three of four predefined measures of tuberculosis exposure. For each hour sharing room air with the index case, the odds of a positive ELISPOT result increased by 1.05 (95% CI: 1.02-1.09, p = 0.003). Only four adults were TST-positive and TST results did not correlate with exposure. Thus, ELISPOT, but not TST, suggested quite extensive nosocomial transmission of multidrug-resistant M. tuberculosis after brief exposure. These results help to explain the apparent importance of casual contact for tuberculosis transmission, and may have implications for prevention. PMID- 15130908 TI - Respiratory viral infections are a distinct risk for bronchiolitis obliterans syndrome and death. AB - Bronchiolitis obliterans syndrome (BOS) is the major obstacle to long-term survival after lung transplantation, in part because its pathogenesis is poorly understood and treatment options are limited. To identify unique risk factors for BOS and death, we performed a retrospective cohort study on 259 consecutive adult lung transplant recipients over a 5-year period. The demographic and clinical characteristics of this population were analyzed for an association between BOS or death and potential risk factors, including community-acquired respiratory viral (CARV) infections, acute rejection, and cytomegalovirus pneumonitis. Respiratory syncytial virus, parainfluenza, influenza, and adenovirus accounted for 21 CARV infections. Univariate and multivariate time-dependent Cox regression analyses demonstrated that this CARV group was more likely to develop BOS, death, and death from BOS. Furthermore, these trends were more pronounced in patients with evidence of lower respiratory tract-CARV (lower-CARV) infections. Notably, the CARV and lower-CARV infections were risk factors for BOS, death, and death from BOS distinct from the risk attributable to acute rejection. Identification of CARV and lower-CARV infections as BOS and mortality risk factors has important clinical implications and may provide insight into disease pathogenesis and accelerate the development of novel treatment strategies to modify post-CARV BOS. PMID- 15130909 TI - Regulation of regional lung perfusion by nitric oxide. AB - Improved oxygenation has previously been shown in patients with acute lung injury when ventilated in prone position. We hypothesized that this was due to higher regional production of nitric oxide in dorsocaudal lung regions. We measured nitric oxide synthase mRNA expression and nitric oxide production by citrulline assay in ventral and dorsal lung tissue from patients. In volunteers, regional lung perfusion in prone and supine postures was assessed by single photon emission computed tomography using (99m)Tc macroaggregated albumin before and after inhibition of nitric oxide synthase by N(G)-monomethyl-L-arginine infusion. Nitric oxide synthase mRNA expression and nitric oxide production were significantly higher in dorsal compared with ventral lung regions. In supine posture, lung perfusion was shifted to ventral parts during nitric oxide synthase inhibition, whereas in the prone posture lung perfusion remained unchanged. Our results suggest a role for endogenous nitric oxide in regulation of regional pulmonary perfusion. PMID- 15130910 TI - Muscarinic (M) receptors in coronary circulation: gene-targeted mice define the role of M2 and M3 receptors in response to acetylcholine. AB - OBJECTIVE: Determining the role of specific muscarinic (M) receptor subtypes mediating responses to acetylcholine (ACh) has been limited by the specificity of pharmacological agents. Deletion of the gene for M5 receptors abolished response to ACh in cerebral blood vessels but did not affect dilation of coronary arteries. The goal of this study was to determine the M receptors mediating responses to ACh in coronary circulation using mice deficient in M2 or M3 receptors (M2-/-, M3-/-, respectively). METHODS AND RESULTS: Coronary arteries from respective wild-type, M2-/-, or M3-/- mice were isolated, cannulated, and pressurized. Diameter was measured with video microscopy. After preconstriction with U46619, ACh produced dose-dependent dilation of coronary arteries that was similar in wild-type and M2-/- mice. In contrast, dilation of coronary arteries from M3-/- mice to ACh was reduced by approximately 80% compared with wild type. The residual response to ACh was atropine insensitive. Relaxation of coronary arteries to other stimuli was similar in M2-/- and M3-/- mice. Similar results were obtained in aorta rings. CONCLUSIONS: These findings provide the first direct evidence that relaxation to ACh in coronary circulation is mediated predominantly by activation of M3 receptors. PMID- 15130911 TI - Mer receptor tyrosine kinase signaling participates in platelet function. AB - OBJECTIVE: Recently, mice made deficient in growth arrest-specific gene 6 product (Gas6) or in which Gas6 gene expression was inhibited were shown to have platelet dysfunction and to be less susceptible to thrombosis. The aim of this study was to define and characterize the relevant Gas6 receptor or receptors involved in platelet function. METHODS AND RESULTS: Using RT-PCR and Western blot analysis we found that mer was the predominantly expressed subtype in mouse and human platelets, whereas axl and rse were not detected. We generated mer-deficient mice by targeted disruption of the mer receptor gene. Platelets derived from mer deficient mice had decreased platelet aggregation in responses to low concentrations of collagen, U46619, and PAR4 thrombin receptor agonist peptide in vitro. However, the response to ADP was not different from wild-type platelets. Knockout of the mer gene protected mice from collagen/epinephrine-induced pulmonary thromoembolism and inhibited ferric chloride-induced thrombosis in vivo. Tail bleeding times, coagulation parameters, and peripheral blood cell counts in mer-deficient mice were similar to wild-type mice. CONCLUSIONS: Our data provide the first evidence that mer, presumably through activation by its ligand Gas6, participates in regulation of platelet function in vitro and platelet-dependent thrombosis in vivo. PMID- 15130912 TI - Basic fibroblast growth factor-induced endothelial proliferation and NO synthesis involves inward rectifier K+ current. AB - OBJECTIVE: Inward rectifier K+ currents (K(ir)) determine the resting membrane potential and thereby modulate essential Ca2+-dependent pathways, like cell growth and synthesis of vasoactive agents in endothelial cells. Basic fibroblast growth factor (bFGF) acts as a vasodilatator and angiogenic factor. Therefore, we investigated the effect of bFGF on K(ir) and assessed the role in proliferation and nitric oxide (NO) formation of endothelial cells. METHODS AND RESULTS: Using the patch-clamp technique, we found characteristic K(ir) in human umbilical cord vein endothelial cells (HUVEC), which were dose-dependently blocked by barium (10 to 100 micromol/L). Perfusion with bFGF (50 ng/mL) caused a significant increase of K(ir), which was blocked by 100 micromol/L barium (n=18, P<0.01). The bFGF induced HUVEC proliferation was significantly inhibited when using 50 to 100 micromol/L barium (n=6; P<0.01). NO production was examined using a cGMP radioimmunoassay. bFGF caused a significant increase of cGMP levels (n=10; P<0.05), which were blocked by barium. CONCLUSIONS: Modulation of K(ir) plays an important role in bFGF-mediated endothelial cell growth and NO formation. PMID- 15130913 TI - Cyclophilin A is a proinflammatory cytokine that activates endothelial cells. AB - OBJECTIVE: Cyclophilin A (CyPA) is an abundant intracellular protein that is considered to be the main target of the immunosuppressive drug cyclosporine A. We and others showed that CyPA is secreted from smooth muscle cells and macrophages in response to oxidative stress and lipopolysaccharide, suggesting a role for CyPA in inflammation. We therefore studied the proinflammatory effects of CyPA on vascular endothelium. METHODS AND RESULTS: Because atherosclerosis is an inflammatory disease, we studied expression of CyPA in atherosclerotic plaques from the ApoE-/- mouse. Using immunohistochemistry, we showed that CyPA was highly expressed in these plaques. Because endothelial cells (EC) are important mediators of inflammation, we next studied the ability of CyPA to activate EC. Human recombinant CyPA activated mitogen-activated protein kinases, including ERK1/2, JNK, and p38 in cultured human umbilical vein EC. CyPA also stimulated IkappaB-alpha phosphorylation and NF-kappaB activation, and induced expression of adhesion molecules including E-selectin and vascular cell adhesion molecule-1. Furthermore, the combination of CyPA and cycloheximide induced EC apoptosis similar to the proapoptotic effect of tumor necrosis factor-alpha. CONCLUSIONS: Our data indicate that CyPA has proinflammatory effects on EC and may play an important role in the pathogenesis of inflammatory diseases, such as atherosclerosis. PMID- 15130914 TI - Human smooth muscle cell subpopulations differentially accumulate cholesteryl ester when exposed to native and oxidized lipoproteins. AB - BACKGROUND: Vascular smooth muscle cells (SMCs) manifest diverse phenotypes and emerging evidence suggests this is caused by inherently distinct SMC subtypes. Recently, Li et al (Circ Res 2001;89:517-525) successfully cloned 2 uniquely responsive SMC subpopulations from a single human artery and we used this unique resource to test the hypothesis that distinct SMC subtypes are differential precursors of foam cell formation. METHODS AND RESULTS: When challenged with human atherogenic native or oxidized hypertriglyceridemic very-low-density lipoprotein (HTG-VLDL), the larger, slower-growing, spindle-shaped HITB5 SMC clone accumulated significantly more cholesteryl ester (CE) and triglyceride (TG) than the smaller, faster-growing epithelioid-shaped HITA2 SMC clone (10 versus 2 microg CE/mg cell protein [PN] and 60 versus 7 microg TG/mg PN, P<0.05). Lipoprotein lipase (LPL), a key enzyme involved in lipoprotein uptake, was identified as one differentially expressed protein that altered the predisposition of HITA2 SMCs for lipid accumulation. Although HITB5 SMCs secreted significantly more LPL than did HITA2 SMCs (0.7 versus 0.2 U/mL media, P<0.05), the addition of bovine milk LPL to HITA2 SMCs, significantly increased native and oxidized HTG-VLDL-induced lipid accumulation. CONCLUSIONS: Inherently distinct SMC subsets are differentially predisposed to lipoprotein-induced lipid accumulation. Moreover, the environment can influence the response of SMC subsets to atherogenic lipoproteins. PMID- 15130915 TI - Infrarenal aortic diameter predicts all-cause mortality. AB - OBJECTIVE: To assess the relationship between infrarenal aortic diameter and subsequent all-cause mortality in men aged 65 years or older. METHODS AND RESULTS: Aortic diameter was measured using ultrasound in 12 203 men aged 65 to 83 years as part of a trial of screening for abdominal aortic aneurysms. A range of cardiovascular risk factors was also documented. Mortality over the next 3 to 7 years was assessed using record linkage. Initial aortic diameter was categorized into 10 intervals, and the relationship between increasing diameter and subsequent mortality was explored using Cox proportional hazard models. Median diameter increased from 21.4 mm in the youngest men to 22.1 mm in the oldest men. The cumulative all-cause mortality increased in a graded fashion with increasing aortic diameter. Using the diameter interval 19 to 22 mm as the reference, the adjusted hazard ratio for all-cause mortality increased from 1.26 (95% CI: 1.09, 1.44; P=0.001) for aortic diameters of 23 to 26 mm to 2.38 (95% CI: 1.22, 4.61; P=0.011) for aortic diameters of 47 to 50 mm. Analysis of causes of death indicated that cardiovascular disease was an important contributor to this increase. CONCLUSIONS: Infrarenal aortic diameter is an independent marker of subsequent all-cause mortality. PMID- 15130916 TI - Should progestins be blamed for the failure of hormone replacement therapy to reduce cardiovascular events in randomized controlled trials? AB - Many observational studies and experimental and animal studies have demonstrated that estrogen replacement therapy (ERT) or hormone replacement therapy (HRT) (estrogen plus progestin) significantly reduces the risk of coronary heart disease. Nonetheless, recent randomized controlled trials demonstrated some trends toward an increased risk of cardiovascular events rather than a reduction of risk. Recently, both the HRT and ERT arms of the Women's Health Initiative (WHI) study were terminated early because of an increased/no incidence of invasive breast cancer, increased incidence of stroke, and increased trend/no protective effects of cardiovascular disease. We discuss the controversial effects of HRT and ERT on cardiovascular system and provide a hypothesis that the failure of HRT and ERT in reducing the risk of cardiovascular events in postmenopausal women might be because of the stage of their atherosclerosis at the time of initiation of HRT or ERT. PMID- 15130917 TI - Deciphering regulatory patterns of inflammatory gene expression from interleukin 1-stimulated human endothelial cells. AB - OBJECTIVE: Endothelial cells comprise a key component of the inflammatory response. We set out to obtain a comprehensive overview of the immediate-early to early gene expression program of interleukin-1 (IL-1)-stimulated endothelial cells and to identify novel transcription factors and regulatory elements. METHODS AND RESULTS: Human umbilical vein endothelial cells (HUVECs) were stimulated with IL-1 for 0, 0.5, 1, 2.5, and 6 hours and analyzed using Affymetrix U133 microarrays. A total of 137 genes were found to be regulated >4 fold, including 18 transcription factors. The expression of selected genes was confirmed by real-time polymerase chain reaction. Cluster analysis was performed in order to group genes according to their expression profiles. To identify novel transcription factor-binding sites, the corresponding promoters were extracted from databases and analyzed for regulatory elements that were over-represented in specific clusters. Several potentially novel DNA binding sites were identified, and one was shown to specifically bind an IL-1-inducible protein from HUVEC. CONCLUSIONS: These results demonstrate that in the early phase after stimulation, IL-1 evokes a complex gene expression program that includes positive but also negative (feedback) regulators of diverse endothelial cell functions. Furthermore, the identification of a new promoter regulatory element demonstrates the feasibility of the bioinformatics-driven approach to discover novel regulatory mechanisms. PMID- 15130918 TI - Intracellular cholesterol transport. AB - Intracellular cholesterol transport is essential for the maintenance of cholesterol homeostasis. Many aspects of cholesterol metabolism are well-known, including its synthesis in the endoplasmic reticulum, its extracellular transport in plasma lipoproteins, its uptake by the low-density lipoprotein receptor, and its regulation of SREBP and LXR transcription factors. These fundamental pathways in cholesterol metabolism all rely on its proper intracellular distribution among subcellular organelles and the plasma membrane. Transport involving the ER and endosomes is essential for cholesterol synthesis, uptake, and esterification, whereas cholesterol catabolism by enzymes in mitochondria and ER generates steroids, bile acids, and oxysterols. Cholesterol is a highly hydrophobic lipid that requires specialized transport in the aqueous cytosol, involving either vesicles or nonvesicular mechanisms. The latter includes hydrophobic cavity transporters such as StAR-related lipid transfer (START) proteins. Molecular understanding of intracellular cholesterol trafficking has lagged somewhat behind other aspects of cholesterol metabolism, but recent advances have defined some transport pathways and candidate proteins. In this review, we discuss cholesterol transport among specific intracellular compartments, emphasizing the relevance of these pathways to cholesterol homeostasis. PMID- 15130919 TI - DC electric fields induce distinct preangiogenic responses in microvascular and macrovascular cells. AB - OBJECTIVE: Electrical stimulation induces significant angiogenesis in vivo. We have shown recently that electrical stimulation induces directional migration, reorientation, and elongation of macrovascular endothelial cells. Because angiogenesis occurs mainly in the microvasculature, we have extended this observation to include human microvascular endothelial cells (HMEC-1s) and compared the responses with that of vascular fibroblasts and smooth muscle cells and human umbilical vein endothelial cells. METHODS AND RESULTS: Four types of vascular cells were cultured in electric fields (EFs). Dynamic cell behaviors were recorded and analyzed with an image analyzer. EFs of 150 to 400 mV/mm induced directed migration, reorientation, and elongation of all the vascular cells. HMEC-1s showed the greatest directional migration (migration rate of 11 microm/h and directedness of 0.35 at 200 mV/mm). Most intriguingly, HMEC-1s migrated toward the cathode, whereas the other cell types migrated toward the anode. CONCLUSIONS: Endothelial cells derived from angiogenic microvascular as opposed to nonangiogenic macrovascular tissues were more responsive to electrical stimulation. This intriguing directional selectivity indicates that a DC electrical signal as a directional cue may be able to play a role in the spatial organization of vascular structure. PMID- 15130920 TI - Angiotensin II stimulates the release of interleukin-6 and interleukin-8 from cultured human adipocytes by activation of NF-kappaB. AB - OBJECTIVE: Several proinflammatory cytokines including IL-6 and IL-8 are produced by human adipocytes, but it is still unclear how this process is regulated. Angiotensin (Ang) II, which is also produced by adipocytes, might play a role as a regulator. In the present study, we investigated the effect of Ang II on the production of IL-6 and IL-8 in in vitro differentiated human adipocytes. METHODS AND RESULTS: Isolation of preadipocytes and differentiation of these cells into adipocytes, Real-time quantitative reverse-transcriptase polymerase chain reaction, Western-blot, enzyme-linked immunosorbent assay, and electromobility shift assay. Ang II-stimulated IL-6 and IL-8 mRNA expression and protein release in a time- and concentration-dependent way. This action of Ang II was completely blocked by the NF-kappaB-blocker Bay 117082 and the AT1 blocker candesartan, but only partially by the AT2-blocker PD 123 319. Incubation of adipocytes with Ang II resulted in an increased phosphorylation of the p65 subunit of NF-kappaB and an increased translocation of NF-kappaB to the nucleus. CONCLUSIONS: Ang II stimulates IL-6 and IL-8 production and release from human adipocytes by a NF kappaB-dependent pathway. This proinflammatory action of Ang II seems to be mediated by the AT1 and less by the AT2 receptor subtype. PMID- 15130921 TI - Small GTP-binding protein Ral is involved in cAMP-mediated release of von Willebrand factor from endothelial cells. AB - OBJECTIVE: von Willebrand factor (vWF) is synthesized by endothelial cells and stored in specialized vesicles called Weibel-Palade bodies (WPBs). Recently, we have shown that the small GTP-binding protein Ral is involved in thrombin-induced exocytosis of WPBs. In addition to Ca2+-elevating secretagogues such as histamine and thrombin, release of WPB is also observed after administration of cAMP raising substances such as epinephrine and vasopressin. In the present study, we investigated whether Ral is also involved in cAMP-mediated vWF release. METHODS AND RESULTS: Activation of Ral was observed 15 to 20 minutes after stimulation of endothelial cells with epinephrine, forskolin, or dibutyryl-cAMP. A cell permeable peptide comprising the carboxy-terminal part of the Ral protein reduced both thrombin-induced and epinephrine-induced vWF secretion supporting a crucial role for Ral in this process. Furthermore, inhibition of protein kinase A by H-89 resulted in a marked reduction of vWF release and greatly diminished levels of GTP-Ral on stimulation with epinephrine. Activation of Ral was independent of the activation of Epac, a cAMP-regulated exchange factor for the small GTPases Rap1 and Rap2. CONCLUSIONS: These results suggest that protein kinase A-dependent activation of Ral regulates cAMP-mediated exocytosis of WPB in endothelial cells. PMID- 15130922 TI - CCL11 (Eotaxin) induces CCR3-dependent smooth muscle cell migration. AB - OBJECTIVE: CCL11 (Eotaxin) is a potent eosinophil chemoattractant that is abundant in atheromatous plaques. The major receptor for CCL11 is CCR3, which is found on leukocytes and on some nonleukocytic cells. We sought to determine whether vascular smooth muscle cells (SMCs) possessed functional CCR3. METHODS AND RESULTS: CCR3 mRNA (by RT-PCR) and protein (by Western blot analysis and flow cytometry) were present in mouse aortic SMCs. CCL11 induced concentration dependent SMC chemotaxis in a modified Boyden chamber, with maximum effect seen at 100 ng/mL. SMC migration was markedly inhibited by antibody to CCR3, but not to CCR2. CCL11 also induced CCR3-dependent SMC migration in a scrape-wound assay. CCL11 had no effect on SMC proliferation. CCR3 and CCL11 staining were minimal in the normal arterial wall, but were abundant in medial SMC and intimal SMC 5 days and 28 days after mouse femoral arterial injury, respectively, times at which SMCs possess a more migratory phenotype. CONCLUSIONS: These data demonstrate that SMCs possess CCR3 under conditions associated with migration and that CCL11 is a potent chemotactic factor for SMCs. Because CCL11 is expressed abundantly in SMC rich areas of the atherosclerotic plaque and in injured arteries, it may play an important role in regulating SMC migration. PMID- 15130923 TI - Analyzing time series gene expression data. AB - MOTIVATION: Time series expression experiments are an increasingly popular method for studying a wide range of biological systems. However, when analyzing these experiments researchers face many new computational challenges. Algorithms that are specifically designed for time series experiments are required so that we can take advantage of their unique features (such as the ability to infer causality from the temporal response pattern) and address the unique problems they raise (e.g. handling the different non-uniform sampling rates). RESULTS: We present a comprehensive review of the current research in time series expression data analysis. We divide the computational challenges into four analysis levels: experimental design, data analysis, pattern recognition and networks. For each of these levels, we discuss computational and biological problems at that level and point out some of the methods that have been proposed to deal with these issues. Many open problems in all these levels are discussed. This review is intended to serve as both, a point of reference for experimental biologists looking for practical solutions for analyzing their data, and a starting point for computer scientists interested in working on the computational problems related to time series expression analysis. PMID- 15130924 TI - WebSIDD: server for predicting stress-induced duplex destabilized (SIDD) sites in superhelical DNA. AB - SUMMARY: WebSIDD is a Web-based service designed to predict locations and extents of stress-induced duplex destabilization (SIDD) that occur in a double-stranded DNA molecule of specified base sequence, on which a specified level of superhelical stress is imposed. The algorithm calculates the approximate equilibrium statistical mechanical distribution of a population of identical molecules among its accessible states. The user inputs the DNA sequence, and the program outputs the calculated transition probability and destabilization energy of each base pair in the sequence. As options, the user can specify the temperature and the level of superhelicity. The values of all structural and energy parameters used in the calculation have been experimentally measured. WebSIDD should prove useful for finding SIDD-susceptible sites in genomic sequences, and correlating their occurrence with locations involved in regulatory and pathological processes. This strategy already has illuminated the roles of SIDD in diverse biological regulatory processes, including transcriptional initiation and termination, and the eukaryotic nuclear scaffold attachments that partition chromosomes into domains. AVAILABILITY: http://orange.genomecenter.ucdavis.edu/benham/sidd/index.html PMID- 15130925 TI - SWAPSC: sliding window analysis procedure to detect selective constraints. AB - Sliding-window analysis procedure to detect selective constraints (SWAPSC) is a software system to dissect the constraints on the evolution of protein-coding genes. The program estimates rates of nucleotide substitutions at specific codon regions in each branch of a phylogenetic tree. The program uses several sets of simulated sequence alignments to estimate the probability of synonymous and non synonymous nucleotide substitutions. Thereafter, a statistical analysis is conducted to determine the optimum window size to detect selective constraints. Finally, the optimum window size is slid along the real alignment and a test for significance of the estimated number of synonymous and non-synonymous nucleotide substitutions in each sliding step is conducted. A number of friendly useful output files is generated. AVAILABILITY: SWAPSC is available at http://www.may.ie/academic/biology/staff/mfmolecevolandbioinf.shtml distribution versions for both Linux and Windows operating systems are available, including manual and example files. PMID- 15130926 TI - LUCY2: an interactive DNA sequence quality trimming and vector removal tool. AB - Lucy2 is a raw DNA sequence trimming and visualization tool based on the popular command-line Lucy1. Users can change parameters, trim multiple sequences and visualize the results within an integrated, easy-to-use graphical user interface. Lucy2 is designed specifically for non-programmers to use, and is currently available on Windows, Linux and MacOS X. Source code is also available for porting to the other platforms. AVAILABILITY: Lucy2 is distributed under the GNU General Public License and can be downloaded from www.complex.iastate.edu PMID- 15130927 TI - Heterogeneity detector: finding heterogeneous positions in Phred/Phrap assemblies. AB - A modification to Phred and program to detect heterogeneous positions, which is particularly useful in the identification of mutations and other abnormalities in Phred/Phrap genome assemblies. AVAILABILITY: The package is made available at http://glscompute.gis.a-star.edu.sg/~charlie/DHetero.html PMID- 15130928 TI - TargetDB: a target registration database for structural genomics projects. AB - TargetDB is a centralized target registration database that includes protein target data from the NIH structural genomics centers and a number of international sites. TargetDB, which is hosted by the Protein Data Bank (RCSB PDB), provides status information on target sequences and tracks their progress through the various stages of protein production and structure determination. A simple search form permits queries based on contributing site, target ID, protein name, sequence, status and other data. The progress of individual targets or entire structural genomics projects may be tracked over time, and target data from all contributing centers may also be downloaded in the XML format. AVAILABILITY: TargetDB is available at http://targetdb.pdb.org/ PMID- 15130929 TI - A collaborative database of inbred mouse strain characteristics. AB - A database and website (MPD: Mouse Phenome Database) have been developed to serve as a consolidated home for mouse strain characterization data being generated by the scientific community. Physiological, anatomical and behavioral data are being collected and integrated into a common framework for tabulation by strain and sex. Genotypic data are being collected as well. The current focus is on a set of 40 inbred strains. The MPD as of February 2004 contains approximately 500 phenotypic parameters relevant to human health, voluntarily contributed by several dozen investigators and laboratories. AVAILABILITY: www.jax.org/phenome PMID- 15130930 TI - Eigenvalue analysis of amino acid substitution matrices reveals a sharp transition of the mode of sequence conservation in proteins. AB - The pattern of amino acid substitutions and sequence conservation over many structure-based alignments of protein sequences was analyzed as a function of percentage sequence identity. The statistics of the amino acid substitutions were converted into the form of log-odds amino acid substitution matrices to which eigenvalue decomposition was applied. It was found that the most important component of the substitution matrices exhibited a sharp transition at the sequence identity of 30-35%, which coincides with the twilight zone. Above the transition point, the most dominant component is related to the mutability of amino acids and it acts to disfavor any substitutions, whereas below the transition point, the most dominant component is related to the hydrophobicity of amino acids and substitutions between residues of similar hydrophobic character are positively favored. Implications for protein evolution and sequence analysis are discussed. PMID- 15130931 TI - ASIAN: a website for network inference. AB - We constructed a website for inferring a network by applying the graphical Gaussian model, from a large amount of data, including redundant information. The available tools on the website are based on a system, named ASIAN (Automatic System for Inferring A Network), in combination with the two methods in our previous papers, which were designed to analyze gene expression profiles on a genomic scale. One of the remarkable features of the website is its ability to infer a network, concomitant with hierarchical clustering and the following estimation of cluster boundaries. AVAILABILITY: http://eureka.ims.u tokyo.ac.jp/asian PMID- 15130932 TI - THEA: ontology-driven analysis of microarray data. AB - MOTIVATION: Microarray technology makes it possible to measure thousands of variables and to compare their values under hundreds of conditions. Once microarray data are quantified, normalized and classified, the analysis phase is essentially a manual and subjective task based on visual inspection of classes in the light of the vast amount of information available. Currently, data interpretation clearly constitutes the bottleneck of such analyses and there is an obvious need for tools able to fill the gap between data processed with mathematical methods and existing biological knowledge. RESULTS: THEA (Tools for High-throughput Experiments Analysis) is an integrated information processing system allowing convenient handling of data. It allows to automatically annotate data issued from classification systems with selected biological information coming from a knowledge base and to either manually search and browse through these annotations or automatically generate meaningful generalizations according to statistical criteria (data mining). AVAILABILITY: The software is available on the website http://thea.unice.fr/ PMID- 15130933 TI - A statistical framework for genomic data fusion. AB - MOTIVATION: During the past decade, the new focus on genomics has highlighted a particular challenge: to integrate the different views of the genome that are provided by various types of experimental data. RESULTS: This paper describes a computational framework for integrating and drawing inferences from a collection of genome-wide measurements. Each dataset is represented via a kernel function, which defines generalized similarity relationships between pairs of entities, such as genes or proteins. The kernel representation is both flexible and efficient, and can be applied to many different types of data. Furthermore, kernel functions derived from different types of data can be combined in a straightforward fashion. Recent advances in the theory of kernel methods have provided efficient algorithms to perform such combinations in a way that minimizes a statistical loss function. These methods exploit semidefinite programming techniques to reduce the problem of finding optimizing kernel combinations to a convex optimization problem. Computational experiments performed using yeast genome-wide datasets, including amino acid sequences, hydropathy profiles, gene expression data and known protein-protein interactions, demonstrate the utility of this approach. A statistical learning algorithm trained from all of these data to recognize particular classes of proteins- membrane proteins and ribosomal proteins--performs significantly better than the same algorithm trained on any single type of data. AVAILABILITY: Supplementary data at http://noble.gs.washington.edu/proj/sdp-svm PMID- 15130934 TI - GO-Mapper: functional analysis of gene expression data using the expression level as a score to evaluate Gene Ontology terms. AB - MOTIVATION: Retrieval of information on biological processes from large-scale expression data is still a time-consuming task. An automated analysis utilizing all expression information would greatly increase our understanding of the samples under study. RESULTS: We describe here a novel method to obtain a functional analysis of complex gene expression data. Instead of applying a predefined expression threshold, Gene Ontology (GO) terms are weighted using the actual measured levels of expression of all associated genes. Based on this concept, the application GO-Mapper was developed to quantitatively link gene expression levels to GO-terms for multiple experiments in an automated way. The applicability of GO-Mapper was developed and validated on in house and public human microarray data and mouse SAGE data. We demonstrate that the GO-Mapper allows for interrelating relevant biological functions with the experiments under study. AVAILABILITY: The GO-Mapper application is free of charge available from our website. PMID- 15130935 TI - A dynamically growing self-organizing tree (DGSOT) for hierarchical clustering gene expression profiles. AB - MOTIVATION: The increasing use of microarray technologies is generating large amounts of data that must be processed in order to extract useful and rational fundamental patterns of gene expression. Hierarchical clustering technology is one method used to analyze gene expression data, but traditional hierarchical clustering algorithms suffer from several drawbacks (e.g. fixed topology structure; mis-clustered data which cannot be reevaluated). In this paper, we introduce a new hierarchical clustering algorithm that overcomes some of these drawbacks. RESULT: We propose a new tree-structure self-organizing neural network, called dynamically growing self-organizing tree (DGSOT) algorithm for hierarchical clustering. The DGSOT constructs a hierarchy from top to bottom by division. At each hierarchical level, the DGSOT optimizes the number of clusters, from which the proper hierarchical structure of the underlying dataset can be found. In addition, we propose a new cluster validation criterion based on the geometric property of the Voronoi partition of the dataset in order to find the proper number of clusters at each hierarchical level. This criterion uses the Minimum Spanning Tree (MST) concept of graph theory and is computationally inexpensive for large datasets. A K-level up distribution (KLD) mechanism, which increases the scope of data distribution in the hierarchy construction, was used to improve the clustering accuracy. The KLD mechanism allows the data misclustered in the early stages to be reevaluated at a later stage and increases the accuracy of the final clustering result. The clustering result of the DGSOT is easily displayed as a dendrogram for visualization. Based on a yeast cell cycle microarray expression dataset, we found that our algorithm extracts gene expression patterns at different levels. Furthermore, the biological functionality enrichment in the clusters is considerably high and the hierarchical structure of the clusters is more reasonable. AVAILABILITY: DGSOT is available upon request from the authors. PMID- 15130936 TI - Distribution of information in biomedical abstracts and full-text publications. AB - MOTIVATION: Full-text documents potentially hold more information than their abstracts, but require more resources for processing. We investigated the added value of full text over abstracts in terms of information content and occurrences of gene symbol--gene name combinations that can resolve gene-symbol ambiguity. RESULTS: We analyzed a set of 3902 biomedical full-text articles. Different keyword measures indicate that information density is highest in abstracts, but that the information coverage in full texts is much greater than in abstracts. Analysis of five different standard sections of articles shows that the highest information coverage is located in the results section. Still, 30-40% of the information mentioned in each section is unique to that section. Only 30% of the gene symbols in the abstract are accompanied by their corresponding names, and a further 8% of the gene names are found in the full text. In the full text, only 18% of the gene symbols are accompanied by their gene names. PMID- 15130937 TI - Incremental generation of summarized clustering hierarchy for protein family analysis. AB - MOTIVATION: Protein sequence clustering has been widely exploited to facilitate in-depth analysis of protein functions and families. For some applications of protein sequence clustering, it is highly desirable that a hierarchical structure, also referred to as dendrogram, which shows how proteins are clustered at various levels, is generated. However, as the sizes of contemporary protein databases continue to grow at rapid rates, it is of great interest to develop some summarization mechanisms so that the users can browse the dendrogram and/or search for the desired information more effectively. RESULTS: In this paper, the design of a novel incremental clustering algorithm aimed at generating summarized dendrograms for analysis of protein databases is described. The proposed incremental clustering algorithm employs a statistics-based model to summarize the distributions of the similarity scores among the proteins in the database and to control formation of clusters. Experimental results reveal that, due to the summarization mechanism incorporated, the proposed incremental clustering algorithm offers the users highly concise dendrograms for analysis of protein clusters with biological significance. Another distinction of the proposed algorithm is its incremental nature. As the sizes of the contemporary protein databases continue to grow at fast rates, due to the concern of efficiency, it is desirable that cluster analysis of a protein database can be carried out incrementally, when the protein database is updated. Experimental results with the Swiss-Prot protein database reveal that the time complexity for carrying out incremental clustering with k new proteins added into the database containing n proteins is O(n2betalogn), where beta congruent with 0.865, provided that k << n. AVAILABILITY: The Linux executable is available on the following supplementary page. PMID- 15130938 TI - Gene co-expression network topology provides a framework for molecular characterization of cellular state. AB - MOTIVATION: Gene expression data have become an instrumental resource in describing the molecular state associated with various cellular phenotypes and responses to environmental perturbations. The utility of expression profiling has been demonstrated in partitioning clinical states, predicting the class of unknown samples and in assigning putative functional roles to previously uncharacterized genes based on profile similarity. However, gene expression profiling has had only limited success in identifying therapeutic targets. This is partly due to the fact that current methods based on fold-change focus only on single genes in isolation, and thus cannot convey causal information. In this paper, we present a technique for analysis of expression data in a graph theoretic framework that relies on associations between genes. We describe the global organization of these networks and biological correlates of their structure. We go on to present a novel technique for the molecular characterization of disparate cellular states that adds a new dimension to the fold-based methods and conclude with an example application to a human medulloblastoma dataset. RESULTS: We have shown that expression networks generated from large model-organism expression datasets are scale-free and that the average clustering coefficient of these networks is several orders of magnitude higher than would be expected for similarly sized scale-free networks, suggesting an inherent hierarchical modularity similar to that previously identified in other biological networks. Furthermore, we have shown that these properties are robust with respect to the parameters of network construction. We have demonstrated an enrichment of genes having lethal knockout phenotypes in the high-degree (i.e. hub) nodes in networks generated from aggregate condition datasets; using process-focused Saccharomyces cerivisiae datasets we have demonstrated additional high-degree enrichments of condition-specific genes encoding proteins known to be involved in or important for the processes interrogated by the microarrays. These results demonstrate the utility of network analysis applied to expression data in identifying genes that are regulated in a state-specific manner. We concluded by showing that a sample application to a human clinical dataset prominently identified a known therapeutic target. AVAILABILITY: Software implementing the methods for network generation presented in this paper is available for academic use by request from the authors in the form of compiled linux binary executables. PMID- 15130939 TI - Human bone marrow megakaryocytes and platelets express PPARgamma, and PPARgamma agonists blunt platelet release of CD40 ligand and thromboxanes. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) is a ligand activated transcription factor important in lipid metabolism, diabetes, and inflammation. We evaluated whether human platelets and megakaryocytes express PPARgamma and whether PPARgamma agonists influence platelet release of bioactive mediators. Although PPARgamma is mainly considered a nuclear receptor, we show that enucleate platelets highly express PPARgamma protein as shown by Western blotting, flow cytometry, and immunocytochemistry. Meg-01 megakaryocyte cells and human bone marrow megakaryocytes also express PPARgamma. Platelet and Meg-01 PPARgamma bound the PPARgamma DNA consensus sequence, and this was enhanced by PPARgamma agonists. Platelets are essential not only for clotting, but have an emerging role in inflammation in part due to their release or production of the proinflammatory and proatherogenic mediators CD40 ligand (CD40L) and thromboxanes (TXs). Platelet incubation with a natural PPARgamma agonist, 15d-PGJ(2), or with a potent synthetic PPARgamma ligand, rosiglitazone, prevented thrombin-induced CD40L surface expression and release of CD40L and thromboxane B(2) (TXB(2)). 15d PGJ(2) also inhibited platelet aggregation and adenosine triphosphate (ATP) release. Our results show that human platelets express PPARgamma and that PPARgamma agonists such as the thiazolidinedione class of antidiabetic drugs have a new target cell, the platelet. This may represent a novel mechanism for treatment of inflammation, thrombosis, and vascular disease in high-risk patients. PMID- 15130940 TI - Dimerization: a versatile switch for oncogenesis. AB - Forced dimerization or oligomerization has emerged as a powerful mechanism for unleashing the oncogenic properties of chimeric transcription factors in acute leukemias. Fusion of transcriptional regulators with a variety of heterologous partner proteins as a consequence of chromosomal rearrangements induces inappropriate self-association, leading to aberrant transcriptional properties and leukemogenesis. Forced dimerization/oligomerization may alter the association of a DNA-binding protein for its transcriptional cofactors, or the dimerization motifs themselves may constitutively recruit transcriptional effector molecules. Oligomerized chimeras may also sequester essential partners or cofactors to exert dominant-negative effects on target gene expression. A key mechanistic feature, and one with major clinical implications, is the nature of the transcriptional cofactors that are recruited by the dimerized oncoprotein. Chimeric RARalpha and acute myeloid leukemia 1 (AML1) proteins induce constitutive repression after the recruitment of corepressors, whereas inappropriate maintenance of target gene expression by mixed-lineage leukemia (MLL) chimeras may result from the recruitment of coactivators or the basal transcriptional machinery. Molecular therapies directed at enzymatic activities of the aberrantly recruited cofactors, or antagonism of dimerization itself, represent promising avenues of current and future investigation. PMID- 15130941 TI - Regulation of monocyte migration by amphoterin (HMGB1). AB - Amphoterin (HMGB1) is a 30-kD heparin-binding protein involved in process extension and migration of cells by a mechanism involving the receptor for advanced glycation end products (RAGE). High levels of amphoterin are released to serum during septic shock. We have studied the expression of amphoterin in monocytes and the role of amphoterin and RAGE in monocyte transendothelial migration. Un-activated monocytes in suspension did not reveal amphoterin on their surface, but adherent monocytes exported amphoterin to the cell surface. Immunohistochemical staining of arterial thrombi in vivo revealed amphoterin in mononuclear cells and in surrounding extracellular matrix. Amphoterin was secreted from phorbol ester and interferon-gamma (IFN-gamma)-activated macrophages, and the secretion was inhibited by blocking the adenosine 5' triphosphate (ATP)-binding cassette transporter-1, a member of the multidrug resistance protein family. Amphoterin was specifically adhesive for monocytes in peripheral blood leukocyte adhesion assay. Adhesion caused an extensive spreading of cells, which was inhibited by the dominant-negative RAGE receptor (soluble ectodomain of RAGE), and adhesion up-regulated chromogranin expression in monocytes, also suggesting a RAGE-dependent interaction. Monocyte transendothelial migration was efficiently inhibited by anti-amphoterin and anti RAGE antibodies and by the soluble RAGE. We suggest that amphoterin is an autocrine/paracrine regulator of monocyte invasion through the endothelium. PMID- 15130942 TI - Exogenous IL-7 increases recent thymic emigrants in peripheral lymphoid tissue without enhanced thymic function. AB - Interleukin 7 (IL-7) is critical in maintaining thymic-dependent and thymic independent pathways of T-cell homeostasis. T-cell receptor (TCR) rearrangement excision circles (TRECs) have been used as markers for recent thymic emigrants (RTEs) in assessing human thymic function. To study the thymic and peripheral effects of IL-7 on RTEs, we measured TREC content and peripheral naive T-cell subsets and turnover in IL-7-treated mice. Short-term administration of IL-7 into thymus-intact mice resulted in increased total TREC numbers, consistent with RTE accumulation. Decreases in TREC frequency were attributable to dilution secondary to increased cell turnover. Significantly, IL-7 administration into thymectomized mice resulted in patterns of decreased TREC frequency and increased total TREC number similar to those in IL-7-treated thymus-intact mice. Distinct patterns of naive cell and RTE distribution among peripheral immune organs and altered expression of CD11a were observed following IL-7 treatment in thymus-intact and thymectomized mice. These results demonstrate (1) that total TREC number and not TREC frequency accurately reflects quantitative changes in RTEs; (2) that short term IL-7 administration results in preferential accumulations of RTEs among peripheral immune organs, accounting for the increase in TRECs in the total peripheral lymphoid pool; and (3) no evidence for regulation of thymic function by short-term IL-7 administration. PMID- 15130943 TI - Bone marrow angiogenic ability and expression of angiogenic cytokines in myeloma: evidence favoring loss of marrow angiogenesis inhibitory activity with disease progression. AB - We compared the angiogenic potential of bone marrow plasma and the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and their receptors on plasma cells from patients with monoclonal gammopathy of undetermined significance (MGUS), smoldering multiple myeloma (SMM), and newly diagnosed multiple myeloma (NMM). Cytokine and cytokine-receptor expression was studied by bone marrow immunohistochemistry, quantitative reverse transcription polymerase chain reaction (RT-PCR) on sorted plasma cells, and quantitative Western blot analysis. Bone marrow angiogenic potential was studied using a human in vitro angiogenesis assay. The expression levels of VEGF, bFGF, and their receptors were similar among MGUS, SMM, and NMM. Sixty-one percent of NMM samples stimulated angiogenesis in the in vitro angiogenesis assay compared with SMM (0%) and MGUS (7%) (P <.001). Importantly, 63% of MGUS samples inhibited angiogenesis compared with SMM (43%) and NMM (4%) (P <.001). The inhibitory activity was heat stable, not overcome by the addition of VEGF, and corresponded to a molecular weight below 10 kd by size-exclusion chromatography. Our results suggest that increasing angiogenesis from MGUS to NMM is, at least in part, explained by increasing tumor burden rather than increased expression of VEGF/bFGF by individual plasma cells. The active inhibition of angiogenesis in MGUS is lost with progression, and the angiogenic switch from MGUS to NMM may involve a loss of inhibitory activity. PMID- 15130944 TI - Tachykinins regulate the function of platelets. AB - Evidence has been mounting for peripheral functions for tachykinins, a family of neuropeptides including substance P (SP), neurokinin A, and neurokinin B, which are recognized for their roles in the central and peripheral nervous system. The recent discovery of 4 new members of this family, the endokinins (EKA, B, C, and D), which are distributed peripherally, adds support to the notion that tachykinins have physiologic/endocrine roles in the periphery. In the present study we report a fundamental new function for tachykinins in the regulation of platelet function. We show that SP stimulates platelet aggregation, and underlying this is the intracellular mobilization of calcium and degranulation. We demonstrate the presence of the tachykinin receptors NK1 and NK3 in platelets and present evidence for the involvement of NK1 in SP-mediated platelet aggregation. Platelets were found to contain SP-like immunoreactivity that is secreted upon activation implicating SP-like substances in the autocrine/paracrine regulation of these cells. Indeed, NK1-blocking antibodies inhibited aggregation in response to other agonists. Of particular note is the observation that EKA/B cross-react in the SP immunoassay and are also able to stimulate platelet activation. Together our data implicate tachykinins, specifically SP and EKA/B, in the regulation of platelet function. PMID- 15130945 TI - CD63 tetraspanin slows down cell migration and translocates to the endosomal lysosomal-MIICs route after extracellular stimuli in human immature dendritic cells. AB - We analyzed herein whether members of the tetraspanin superfamily are involved in human immature dendritic cell (DC) functions such as foreign antigen internalization, phagocytosis, and cell migration. We show that CD63, CD9, CD81, CD82, and CD151 are present in immature DCs. Whereas CD9 and CD81 are mostly expressed at the cell surface, CD63 and CD82 are also located in intracellular organelles. Complexes of monoclonal antibody (Mab) FC-5.01-CD63 or Fab-5.01-CD63 were rapidly translocated "outside-in" and followed the endocytic pathway through early endosomes and lysosomes, reaching major histocompatibility complex (MHC) class II-enriched compartments (MIICs) in less than one hour. Internalization of CD63 was also observed during Saccharomyces cerevisiae phagocytosis. Moreover, an association of CD63 with the beta-glycan receptor dectin-1 was observed. Mabs against CD9, CD63, CD81, and CD82 enhanced by 50% the migration induced by the chemokines macrophage inflammatory protein-5 (MIP-5) and MIP-1alpha. Concomitantly, Mabs against CD63 and CD82 diminished the surface expression of CD29, CD11b, CD18, and alpha5 integrins. By immunoprecipitation experiments we found that CD63 associated with integrins CD11b and CD18. These results suggest that CD9, CD63, CD81, and CD82 could play a role in modulating the interactions between immature DCs and their environment, slowing their migratory ability. However, only CD63 would intervene in the internalization of complex antigens. PMID- 15130946 TI - Megakaryocytes express functional Aurora-B kinase in endomitosis. AB - Endomitosis (EnM) in megakaryocytes (MKs) is characterized by abortion of mitosis in late anaphase and failure of cytokinesis; subsequent reinitiation of DNA synthesis results in polyploidy. Ablation of chromosomal passenger proteins including Aurora-B kinase causes defects in late anaphase and cytokinesis in diploid cells; thus one hypothesis is that the expression or function of these proteins in polyploid MKs is abnormal. It has been reported that Aurora-B kinase mRNA is decreased in polyploid megakaryocytic cells, suggesting that deficiency of Aurora-B kinase is responsible for EnM. We examined the localization of Aurora B kinase and additional members of the chromosomal passenger protein and aurora kinase families in MKs. We found that in EnM MKs (1) Aurora-B kinase is present and appropriately localized to centromeres in early EnM; (2) in low-ploidy human MKs, centromeric localization of survivin and inner centromere protein (INCENP) can also be demonstrated; (3) the function of Aurora-B kinase, as measured by Ser10 phosphorylation of histone H3, is intact; and (4) aurora-A kinase localizes appropriately to centrosomes in EnM. These results suggest that EnM MKs appropriately express functional Aurora-B kinase and related proteins in early anaphase, making a simple deficiency of this protein an unlikely explanation for polyploidy in this cell type. PMID- 15130947 TI - Lymphocyte microvilli are dynamic, actin-dependent structures that do not require Wiskott-Aldrich syndrome protein (WASp) for their morphology. AB - Short microvilli cover the surfaces of circulating mammalian lymphocytes. The surfaces of monocytes and neutrophils are very different, containing ruffles as their predominant structure. In this study, we present the first quantitative characterization of lymphocyte microvilli. From analysis of scanning electron micrographs, we find that median microvillar length and surface density range from 0.3 to 0.4 microm and 2 to 4 microvilli/microm(2), respectively, on lymphocytes from a variety of sources. As with similar structures from other cells, lymphocyte microvilli contain parallel bundles of actin filaments. Lymphocyte microvilli rapidly disassemble when exposed to the actin-sequestering molecule, Latrunculin A. This disassembly parallels cellular actin filament depolymerization and is complete within 2 minutes, suggesting that lymphocyte microvilli undergo continuous assembly and disassembly. In contrast to previous reports suggesting lymphocyte microvillar density to be reduced on lymphocytes from Wiskott-Aldrich syndrome (WAS) patient, we find no such deficiency in either mouse or human WAS protein (WASp)-deficient lymphocytes. These results suggest that WASp is either not involved in or is redundant in the rapid dynamics of lymphocyte microvilli. PMID- 15130948 TI - Molecular mechanism of violacein-mediated human leukemia cell death. AB - Violacein, a pigment isolated from Chromobacterium violaceum in the Amazon River, presents diverse biologic properties and attracts interest as a consequence of its antileukemic activity. Elucidation of the molecular mechanism mediating this activity will provide further relevant information for understanding its effects on the cellular physiology of untransformed cells and for considering its possible clinical application. Here, we show that violacein causes apoptosis in HL60 leukemic cells but is ineffective in this respect in other types of leukemia cells or in normal human lymphocytes and monocytes. Violacein cytotoxicity in HL60 cells was preceded by activation of caspase 8, transcription of nuclear factor kappaB (NF-kappaB) target genes, and p38 mitogen-activated protein (MAP) kinase activation. Thus, violacein effects resemble tumor necrosis factor alpha (TNF-alpha) signal transduction in these cells. Accordingly, infliximab, an antibody that antagonizes TNF-alpha-induced signaling abolished the biologic activity of violacein. Moreover, violacein directly activated TNF receptor 1 signaling, because a violacein-dependent association of TNF receptor-associated factor 2 (TRAF2) to this TNF receptor was observed in coimmunoprecipitation experiments. Hence, violacein represents the first member of a novel class of cytotoxic drugs mediating apoptosis of HL60 cells by way of the specific activation of TNF receptor 1. PMID- 15130949 TI - Cortical selective vulnerability in motor neuron disease: a morphometric study. AB - Neuroimaging and neuropsychological studies have revealed that the primary motor cortex (PMC) and the extramotor cortical areas are functionally abnormal in motor neuron disease (MND, amyotrophic lateral sclerosis), but the nature of the cortical lesions that underlie these changes is poorly understood. In particular, there have been few attempts to quantify neuronal loss in the PMC and in other cortical areas in MND. We used SMI-32, an antibody against an epitope on non phosphorylated neurofilament heavy chain, to analyse the size and density of SMI 32-positive cortical pyramidal neurons in layer V of the PMC, the dorsolateral prefrontal cortex (DLPFC) and the supragenual anterior cingulate cortex (ACC) in 13 MND and eight control subjects. There was a statistically significant reduction in the density of SMI-32-immunoreactive (IR) pyramidal neurons within cortical layer V in the PMC, the DLPFC and the ACC in MND subjects compared with controls [t (19) = 2.91, P = 0.009; estimated reduction 25%; 95% CI = 8%, 40%]. In addition, we studied the density and size of interneurons immunoreactive for the calcium-binding proteins calbindin-D(28K) (CB), parvalbumin (PV) and calretinin (CR) in the same areas (PMC, DLPFC and ACC). Statistically significant differences in the densities of CB-IR neurons were observed within cortical layers V (P = 0.003) and VI (P = 0.001) in MND cases compared with controls. The densities of CR- and PV-IR neurons were not significantly different between MND and control cases, although there were trends towards reductions of CR-IR neuronal density within the same layers and of PV-IR neuronal density within cortical layer VI. Loss of pyramidal neurons and of GABAergic interneurons is more widespread than has been appreciated and is present in areas associated with neuroimaging and cognitive abnormalities in MND. These findings support the notion that MND should be considered a multisystem disorder. PMID- 15130950 TI - Pregnancy and multiple sclerosis (the PRIMS study): clinical predictors of post partum relapse. AB - The influence of pregnancy in multiple sclerosis has been a matter of controversy for a long time. The Pregnancy in Multiple Sclerosis (PRIMS) study was the first large prospective study which aimed to assess the possible influence of pregnancy and delivery on the clinical course of multiple sclerosis. We report here the 2 year post-partum follow-up and an analysis of clinical factors which might predict the likelihood of a relapse in the 3 months after delivery. The relapse rate in each trimester up to the end of the second year post-partum was compared with that in the pre-pregnancy year. Clinical predictors of the presence or absence of a post-partum relapse were analysed by logistic regression analysis. Using the best multivariate model, women were classified as having or not having a post-partum relapse predicted, and this was compared with the observed outcome. The results showed that, compared with the pre-pregnancy year, there was a reduction in the relapse rate during pregnancy, most marked in the third trimester, and a marked increase in the first 3 months after delivery. Thereafter, from the second trimester onwards and for the following 21 months, the annualized relapse rate fell slightly but did not differ significantly from the relapse rate recorded in the pre-pregnancy year. Despite the increased risk for the 3 months post-partum, 72% of the women did not experience any relapse during this period. Confirmed disability continued to progress steadily during the study period. Three indices, an increased relapse rate in the pre-pregnancy year, an increased relapse rate during pregnancy and a higher DSS (Kurtzke's Disability Status Scale) score at pregnancy onset, significantly correlated with the occurrence of a post-partum relapse. Neither epidural analgesia nor breast feeding was predictive. When comparing the predicted and observed status, however, only 72% of the women were correctly classified by the multivariate model. In conclusion, the results for the second year post-partum confirm that the relapse rate remains similar to that of the pre-pregnancy year, after an increase in the first trimester following delivery. Women with greater disease activity in the year before pregnancy and during pregnancy have a higher risk of relapse in the post- partum 3 months. This is, however, not sufficient to identify in advance women with multiple sclerosis who are more likely to relapse, especially for planning therapeutic trials aiming to prevent post-partum relapses. PMID- 15130951 TI - Simultaneous neuroprotection and blockade of inflammation reverses autoimmune encephalomyelitis. AB - In multiple sclerosis, the immune system attacks the white matter of the brain and spinal cord, leading to disability and/or paralysis. Myelin, oligodendrocytes and neurons are lost due to the release by immune cells of cytotoxic cytokines, autoantibodies and toxic amounts of the excitatory neurotransmitter glutamate. Experimental autoimmune encephalomyelitis (EAE) is an animal model that exhibits the clinical and pathological features of multiple sclerosis. Current therapies that suppress either the inflammation or glutamate excitotoxicity are partially effective when administered at an early stage of EAE, but cannot block advanced disease. In a multi-faceted approach to combat EAE, we blocked inflammation with an anti-MAdCAM-1 (mucosal addressin cell adhesion molecule-1) monoclonal antibody and simultaneously protected oligodendrocytes and neurons against glutamate mediated damage with the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA)/kainate antagonist 2,3-dihydroxy-6-nitro-7- sulfamoylbenzo(f)quinoxaline (NBQX) and the neuroprotector glycine-proline-glutamic acid (GPE; N-terminal tripeptide of insulin-like growth factor). Remarkably, administration at an advanced stage of unremitting EAE of either a combination of NBQX and GPE, or preferably all three latter reagents, resulted in amelioration of disease and repair of the CNS, as assessed by increased oligodendrocyte survival and remyelination, and corresponding decreased paralysis, inflammation, CNS apoptosis and axonal damage. Each treatment reduced the expression of nitric oxide and a large panel of proinflammatory and immunoregulatory cytokines, in particular IL-6 which plays a critical role in mediating EAE. Mice displayed discernible improvements in all physical features examined. Disease was suppressed for 5 weeks, but relapsed when treatment was suspended, suggesting treatment must be maintained to be effective. The above approaches, which allow CNS repair by inhibiting inflammation and/or simultaneously protect neurons and oligodendrocytes from damage, could thus be effective therapies for multiple sclerosis. PMID- 15130952 TI - Accurate bidirectional saccade control by a single hemicortex. AB - Anatomical, electrophysiological and lesion studies indicate that each cortical hemisphere normally generates saccades directed to the contralateral side. In contrast, in patients who had an entire cortical hemisphere removed surgically (hemidecortication), the remaining hemicortex can generate both contraversive and ipsiversive saccades. However, current evidence indicates that ipsiversive saccades are grossly inaccurate. The obvious reason for this is that hemidecorticate patients are blind in the hemifield ipsilateral to the remaining hemicortex, and therefore normal visual signals are not available to drive ipsiversive saccades. However, absent vision also implies that visual error signals are not available to calibrate ipsiversive movements. Furthermore, the innate anatomical substrate needed to support accurate ipsiversive saccade control, in addition to the normal contraversive control, appears sparse. We show here that, in spite of these obstacles, hemidecorticate patients could generate accurate ipsiversive saccades in a task that dissociated hemianopia from saccade direction. In this task, while the patients fixated a central fixation target (FT), saccade targets (STs) were briefly presented to the intact visual hemifield contralateral to the intact hemicortex. The FT was then moved towards and beyond the former location of the ST which evoked tracking eye movements that moved the eyes towards and then beyond the ST, thereby moving the goal, ST, into the blind visual hemifield ipsilateral to the intact hemicortex. When the FT was extinguished, the patients generated, in the dark, ipsiversive saccades that moved their eyes to the remembered location of the ST with the same accuracy as normal control subjects. This indicates that a single hemicortex can mediate accurate bidirectional saccade control via fully functional bilateral connections from cortex to brainstem oculomotor structures. The mechanisms whereby visual signals can calibrate ipsiversive saccades remain elusive. PMID- 15130953 TI - Hypoxic tissue in ischaemic stroke: persistence and clinical consequences of spontaneous survival. AB - In ischaemic stroke, expansion of the infarct core occurs at the expense of surrounding hypoxic, metabolically compromised tissue over a period of 24 h or more in a considerable proportion of patients. It is uncertain whether hypoxic tissue observed at later times after stroke onset retains the potential for survival or whether such survival has an impact on functional outcome. These factors may determine the effectiveness of therapeutic strategies aimed at salvaging this tissue. We tested the hypotheses that metabolically compromised hypoxic tissue observed within 48 h after onset of ischaemic stroke retains the potential for spontaneous survival and that the impact of such survival on functional outcome is time dependent. Consecutive patients presenting within 48 h of ischaemic stroke were studied with [(18)F]fluoromisonidazole, a ligand binding to hypoxic but viable tissue, and PET. Subjects were grouped into two time epochs, 12 h, based on the interval from stroke onset to the time of tracer injection, and had infarct volumes measured on CT/MRI at 7 days (n = 60). The total ischaemic volume (TIV) and the proportion of the TIV that spontaneously survived (surviving hypoxic volume ratio, SHVR) were defined from the co registered CT/MRI images. These volumetric measures were correlated with neurological outcome assessed at day 7-10 by percentage change in the National Institutes of Health Stroke Scale (DeltaNIHSS), and at 3 months by Barthel Index (BI) and modified Rankin Score (mRS). Of 66 patients investigated, hypoxic tissue occurred in 33 and outcome data was available in 27. Hypoxic tissue constituted >20% of the TIV in 60% of studies 12 h. The spontaneously surviving proportion of the TIV (median 6.9%) or hypoxic tissue (median 45.9%) was not significantly different in patient subgroups studied 12 h after stroke onset. Spontaneous survival of hypoxic tissue (surviving hypoxic volume ratio) was associated with improved neurological outcome in both time epochs: 12 h, DeltaNIHSS (r = 0.59, P < 0.01) and day 90 mRS (r = -0.46, P < 0.05). The finding that similar proportions of hypoxic tissue survived spontaneously within each time epoch suggests that its fate is not predetermined. The favourable neurological outcome associated with spontaneous survival of hypoxic tissue, even 12-48 h after stroke onset, suggests that the volume of hypoxic tissue that progressed to infarction may represent a valuable target for therapeutic intervention. PMID- 15130954 TI - Familial clustering and genetic risk for dementia in a genetically isolated Dutch population. AB - Despite advances in elucidating the genetic epidemiology of Alzheimer's disease and frontotemporal dementia, the aetiology for most patients with dementia remains unclear. We examined the genetic epidemiology of dementia in a recent genetically isolated Dutch population founded around 1750. The series of 191 patients ascertained comprised 122 probable Alzheimer's disease patients with late onset and 17 with early onset, and 22 with possible Alzheimer's disease. It further included 10 patients with vascular dementia, nine with Lewy body dementia and six with frontotemporal dementia. All patients, except those with vascular dementia, were more closely related than healthy individuals from the same area. Clustering was strongest for patients with early-onset Alzheimer's disease or Lewy body dementia. Although 14% of late-onset Alzheimer's disease patients had evidence of autosomal dominant disease, consanguinity was found in three late onset Alzheimer's disease patients, suggesting a recessive or polygenic model underlying the trait. We found no clustering of vascular dementia, implying a difference in genetic risk for late-onset Alzheimer's disease and vascular dementia. Mutations in known genes could not explain the occurrence of dementia, but the population attributable proportion of apolipoprotein E gene (APOE*4) was high (45%) due to a high frequency of APOE*4 carriers. Earlier identified regions on chromosomes 10 and 12, nor the effect of the alpha-2-macroglobulin (A2M) I/D polymorphism on Alzheimer's disease could be confirmed in our study. We did find evidence for association between the A2M D-allele and Lewy body dementia. Our data showed a strong familial clustering of various forms of dementia in this isolated Dutch population. A high percentage of late-onset Alzheimer's disease could be explained by APOE*4, but 55% of its origin is still unknown. PMID- 15130955 TI - Informal care giving for disabled stroke survivors. PMID- 15130956 TI - Roll Back Malaria: a failing global health campaign. PMID- 15130957 TI - National screening programme for aortic aneurysm. PMID- 15130958 TI - Assisted suicide. PMID- 15130959 TI - Physical activity and coronary heart disease. PMID- 15130960 TI - College proposes better deal for acute medical patients. PMID- 15130961 TI - Companies offer surgeons incentives to use their prostheses. PMID- 15130962 TI - Doctors charged with planning to kill a patient for his kidneys. PMID- 15130963 TI - Half of Russia's doctors face sack in healthcare reforms. PMID- 15130964 TI - Newer hypnotics no better for insomnia than short acting benzodiazepines. PMID- 15130965 TI - NICE gives guidance on use of new antiepileptic drugs in children. PMID- 15130966 TI - Open access could reduce cost of scientific publishing. PMID- 15130967 TI - New EU trials database is criticised for lack of openness. PMID- 15130968 TI - US universities get round regulations on stem cell research. PMID- 15130971 TI - Health agencies end in-fighting on malaria. PMID- 15130973 TI - Patients will be able to report drugs' side effects. PMID- 15130974 TI - Mentally ill offenders are being wrongly held in prisons. PMID- 15130975 TI - A lawyer with the Hygeia touch. PMID- 15130976 TI - Some way to go for consistent implementation of guidance on hip fracture. PMID- 15130977 TI - Training carers of stroke patients: randomised controlled trial. AB - BACKGROUND: Informal care givers support disabled stroke patients at home but receive little training for the caregiving role. OBJECTIVE: To evaluate the effectiveness of training care givers in reducing burden of stroke in patients and their care givers. DESIGN: A single, blind, randomised controlled trial. SETTING: Stroke rehabilitation unit. SUBJECTS: 300 stroke patients and their care givers. INTERVENTIONS: Training care givers in basic nursing and facilitation of personal care techniques. MAIN OUTCOME MEASURES: Cost to health and social services, caregiving burden, patients' and care givers' functional status (Barthel index, Frenchay activities index), psychological state (hospital anxiety and depression score), quality of life (EuroQol visual analogue scale) and patients' institutionalisation or mortality at one year. RESULTS: Patients were comparable for age (median 76 years; interquartile range 70-82 years), sex (53% men), and severity of stroke (median Barthel index 8; interquartile range 4-12). The costs of care over one year for patients whose care givers had received training were significantly lower (10,133 pounds sterling v 13,794 pounds sterling (18,087 dollars v 24,619 dollars; 15,204 euros v 20,697 euros); P = 0.001). Trained care givers experienced less caregiving burden (care giver burden score 32 v 41; P = 0.0001), anxiety (anxiety score 3 v 4; P = 0.0001) or depression (depression score 2 v 3; P = 0.0001) and had a higher quality of life (EuroQol score 80 v 70; P = 0.001). Patients' mortality, institutionalisation, and disability were not influenced by caregiver training. However, patients reported less anxiety (3 v 4.5; P < 0.0001) and depression (3 v 4; P < 0.0001) and better quality of life (65 v 60; P = 0.009) in the caregiver training group. CONCLUSION: Training care givers during patients' rehabilitation reduced costs and caregiver burden while improving psychosocial outcomes in care givers and patients at one year. PMID- 15130978 TI - Training care givers of stroke patients: economic evaluation. AB - BACKGROUND: Training care givers reduces their burden and improves psychosocial outcomes in care givers and patients at one year. However, the cost effectiveness of this approach has not been investigated. OBJECTIVE: To evaluate the cost effectiveness of caregiver training by examining health and social care costs, informal care costs, and quality adjusted life years in care givers. DESIGN: A single, blind, randomised controlled trial. SETTING: Stroke rehabilitation unit. SUBJECTS: 300 stroke patients and their care givers. INTERVENTIONS: Caregiver training in basic nursing and facilitation of personal care techniques compared with no care giver training. MAIN OUTCOME MEASURES: Health and social care costs, informal care costs, and quality adjusted life years in care givers over one year after stroke. RESULTS: Total health and social care costs over one year for patients whose care givers received training were significantly lower (mean difference -4043 pounds sterling (7249 dollars; 6072 euros), 95% confidence interval -6544 pounds sterling to -595 pounds sterling). Inclusion of informal care costs, which were similar between the two groups, did not alter this conclusion. The cost difference was largely due to differences in length of hospital stay. The EQ-5D did not detect changes in quality adjusted life years in care givers. CONCLUSION: Compared with no training, caregiver training during rehabilitation of patients reduced costs of care while improving overall quality of life in care givers at one year. PMID- 15130979 TI - Variations in the hospital management of self harm in adults in England: observational study. PMID- 15130980 TI - Management of diastolic heart failure in older adults. PMID- 15130981 TI - Ergonomics in medicine and surgery. PMID- 15130982 TI - Low back pain. PMID- 15130983 TI - Screening for abdominal aortic aneurysms in men. PMID- 15130984 TI - Lessons from developing nations on improving health care. PMID- 15130985 TI - Linking disease control programmes in rural Africa: a pro-poor strategy to reach Abuja targets and millennium development goals. PMID- 15130986 TI - No time to train the surgeons: golden age of surgical training didn't exist. PMID- 15130987 TI - No time to train the surgeons: only repeal of European Working Time Directive will help. PMID- 15130988 TI - No time to train the surgeons: medical profession must re-establish its independence from government. PMID- 15130989 TI - No time to train the surgeons: Royal College of Surgeons of England responds. PMID- 15130990 TI - No time to train the surgeons: the rot is deep. PMID- 15130991 TI - No time to train the surgeons: learning from the New Zealand experience. PMID- 15130992 TI - No time to train the surgeons: choosing trainees carefully may be the solution. PMID- 15130994 TI - Scan immediately for stroke using MRI when possible. PMID- 15130995 TI - Turning a blind eye: testing the success of blinding and the CONSORT statement. PMID- 15130996 TI - Turning a blind eye: authors have blinkered view of blinding. PMID- 15130997 TI - Turning a blind eye: why we don't test for blindness at the end of our trials. PMID- 15130998 TI - We are not "amueslied". PMID- 15130999 TI - Histamine H4 antagonism: a therapy for chronic allergy? PMID- 15131001 TI - Enhanced inhibition of the EDHF phenomenon by a phenyl methoxyalaninyl phosphoramidate derivative of dideoxyadenosine. AB - In rabbit arteries endogenous production of cAMP facilitates electrotonic signalling via gap junctions, thus explaining the ability of P-site inhibitors of adenylyl cyclase to attenuate EDHF-type responses. In the present study, we show that a lipophilic phosphoramidate pronucleotide derivative of dideoxyadenosine, 2',3'-ddA-PMAPh, exhibits enhanced activity as an inhibitor of EDHF-type smooth muscle hyperpolarizations induced by acetylcholine (ACh) compared to the parent nucleoside 2',3'-ddA, and that the effects of both compounds can be reversed by the cAMP phosphodiesterase inhibitor IBMX. Neither 2',3'-ddA nor 2',3'-ddA-PMAPh depress ACh-evoked endothelial hyperpolarization directly. Modifications in the lipophilicity of dideoxyadenosine and its direct intracellular delivery as a mononucleotide may thus enhance the ability to inhibit adenylyl cyclase and depress electrotonic signalling via myoendothelial gap junctions. PMID- 15131000 TI - The complexities of the cardiovascular actions of cannabinoids. AB - The cardiovascular actions of cannbinoids are complex. In general they cause vasorelaxation in isolated blood vessels, while in anaesthetised animals they cause multiphasic responses which involve an early bradycardia and long-lasting hypotension. However, in conscious animals, the picture is one of bradycardia followed by pressor responses. Clearly, the responses to cannabinoids are dependent on the experimental conditions and synthetic cannabinoids and endocannabinoids exhibit different pharmacologies. In terms of mechanisms involved in the vascular responses to cannabinoids, the following have been implicated: the involvement of 'classical' cannabinoid receptors, the involvement of a novel endothelial cannabinoid receptor, the release of nitric oxide, the release of endothelium-derived hyperpolarising factor (EDHF), the activation of vanilloid receptors, metabolism of endocannabinoids to vasoactive molecules, and both peripheral inhibition and central excitation of the sympathetic nervous system. PMID- 15131002 TI - Histamine H4 receptor mediates eosinophil chemotaxis with cell shape change and adhesion molecule upregulation. AB - 1. During mast cell degranulation, histamine is released in large quantities. Human eosinophils were found to express histamine H(4) but not H(3) receptors. The possible effects of histamine on eosinophils and the receptor mediating these effects were investigated in our studies. 2. Histamine (0.01-30 microm) induced a rapid and transient cell shape change in human eosinophils, but had no effects on neutrophils. The maximal shape change was at 0.3 microm histamine with EC(50) at 19 nm. After 60 min incubation with 1 microm histamine, eosinophils were desensitized and were refractory to shape change response upon histamine restimulation. Histamine (0.01-1 microm) also enhanced the eosinophil shape change induced by other chemokines. 3. Histamine-induced eosinophil shape change was mediated by the H(4) receptor. This effect was completely inhibited by H(4) receptor-specific antagonist JNJ 7777120 (IC(50) 0.3 microm) and H(3)/H(4) receptor antagonist thioperamide (IC(50) 1.4 microm), but not by selective H(1), H(2) or H(3) receptor antagonists. H(4) receptor agonists imetit (EC(50) 25 nm) and clobenpropit (EC(50) 72 nm) could mimic histamine effect in inducing eosinophil shape change. 4. Histamine (0.01-100 microm) induced upregulation of adhesion molecules CD11b/CD18 (Mac-1) and CD54 (ICAM-1) on eosinophils. This effect was mediated by the H(4) receptor and could be blocked by H(4) receptor antagonists JNJ 7777120 and thioperamide. 5. Histamine (0.01-10 microm) induced eosinophil chemotaxis with an EC(50) of 83 nm. This effect was mediated by the H(4) receptor and could be blocked by H(4) receptor antagonists JNJ 7777120 (IC(50) 86 nm) and thioperamide (IC(50) 519 nm). Histamine (0.5 microm) also enhanced the eosinophil shape change induced by other chemokines. 6. In conclusion, we have demonstrated a new mechanism of eosinophil recruitment driven by mast cells via the release of histamine. Using specific histamine receptor ligands, we have provided a definitive proof that the H(4) receptor mediates eosinophil chemotaxis, cell shape change and upregulation of adhesion molecules. The effect of H(4) receptor antagonists in blocking eosinophil infiltration could be valuable for the treatment of allergic diseases. The histamine-induced shape change and upregulation of adhesion molecules on eosinophils can serve as biomarkers for clinical studies of H(4) receptor antagonists. PMID- 15131006 TI - High blood pressure upregulates arterial L-type Ca2+ channels: is membrane depolarization the signal? AB - Long-lasting Ca2+ (Ca(L)) channels of the Ca(v)1.2 gene family contribute to the pathogenesis of abnormal arterial tone in hypertension. The physiological stimulus that enhances Ca(L) channel current in the vascular smooth muscle cells (VSMCs) remains unknown. The present study investigated if high blood pressure triggers an upregulation of vascular Ca(L) channel protein. Rat aortae were banded between the origins of the left renal (LR) and right renal (RR) arteries to selectively elevate blood pressure in the proximal RR arteries. After 2 days, the immunoreactivity on Western blots corresponding to the pore-forming alpha1C subunit of the Ca(L) channel was increased 3.25-fold in RR compared with LR arteries. This finding persisted at 28 days and was associated with abnormal Ca2+ dependent tone and higher Ca(L) currents in the VSMCs exposed to high pressure. Based on microelectrode studies indicating that RR arteries were depolarized compared with LR arteries, further studies examined if membrane depolarization, an inherent response of VSMCs to high blood pressure, increased alpha1C expression. Isolated rat renal arteries were cultured for 2 days in low K+ (4 mmol/L) or depolarizing high K+ (30 mmol/L) media. Arteries preconditioned in high K+ showed a 5.47-fold increase in alpha1C expression, enhanced Ca(L) channel current, and elevated Ca2+-dependent tone. These findings provide the first direct evidence that high blood pressure upregulates the Ca(L) channel alpha1C subunit in VSMCs in vivo and suggest that membrane depolarization is a potential signal involved in this interaction that may contribute to the development of abnormal vascular tone. PMID- 15131005 TI - Cardiac angiotensin II participates in coronary microvessel inflammation of unstable angina and strengthens the immunomediated component. AB - Angiotensin (Ang) II is now recognized to be a mediator of a wide variety of inflammatory processes. This study investigated renin-angiotensin system (RAS) components and a number of inflammatory mediators in left ventricular biopsies from 2-vessel disease unstable angina (UA) (n=43) and stable angina (SA) (n=15) patients undergoing coronary bypass surgery. Biopsy samples from 6 patients undergoing valve replacement for mitral stenosis served as controls. UA patients were randomly assigned to angiotensin-converting enzyme (ACE)-inhibitor (ramipril), AT1 antagonist (valsartan), or placebo and treated during the 5 days preceding coronary bypass surgery, performed from 6 to 9 days after coronary angiography. During coronary angiography coronary blood flow was measured and samples were obtained from aorta and coronary sinus for determination of Ang I and Ang II gradients. The hearts of UA patients produced Ang II in a greater amount than in SA patients (P<0.01). UA biopsy samples showed numerous DR+ cells, identified as lymphocytes, macrophages, and endothelial cells. Reverse transcriptase polymerase chain reaction showed overexpression of AGTN, ACE, and AT1-R genes, as well as upregulation of TNF-alpha, IL-6, IFN-gamma, and iNOS genes (P<0.01), with no differences between nonischemic and potentially ischemic areas. AGTN, ACE, and cytokine genes were mainly localized on endothelial cells. Ramipril and valsartan markedly decreased the expression levels of TNF-alpha, IL 6, and iNOS, and, to a lesser extent, of IFN-gamma genes, but did not affect the number of DR+ cells, with no significant difference between the 2 treatments. These results show that locally generated Ang II amplifies the immunomediated inflammatory process of coronary microvessels occurring in unstable angina. PMID- 15131007 TI - Proteomic and metabolomic analysis of vascular smooth muscle cells: role of PKCdelta. AB - Recent developments of proteomic and metabolomic techniques provide powerful tools for studying molecular mechanisms of cell function. Previously, we demonstrated that neointima formation was markedly increased in vein grafts of PKCdelta-deficient mice compared with wild-type controls. To clarify the underlying mechanism, we performed a proteomic and metabolomic analysis of cultured vascular smooth muscle cells (SMCs) derived from PKCdelta+/+ and PKCdelta-/- mice. Using 2-dimensional electrophoresis and mass spectrometry, we identified >30 protein species that were altered in PKCdelta-/- SMCs, including enzymes related to glucose and lipid metabolism, glutathione recycling, chaperones, and cytoskeletal proteins. Interestingly, nuclear magnetic resonance spectroscopy confirmed marked changes in glucose metabolism in PKCdelta-/- SMCs, which were associated with a significant increase in cellular glutathione levels resulting in resistance to cell death induced by oxidative stress. Furthermore, PKCdelta-/- SMCs overexpressed RhoGDIalpha, an endogenous inhibitor of Rho signaling pathways. Inhibition of Rho signaling was associated with a loss of stress fiber formation and decreased expression of SMC differentiation markers. Thus, we performed the first combined proteomic and metabolomic study in vascular SMCs and demonstrate that PKCdelta is crucial in regulating glucose and lipid metabolism, controlling the cellular redox state, and maintaining SMC differentiation. PMID- 15131008 TI - Flow-dependent dilation mediated by endogenous kinins requires angiotensin AT2 receptors. AB - The vascular kallikrein-kinin system contributes to about one third of flow dependent dilation in mice carotid arteries, by activating bradykinin B2 receptors coupled to endothelial nitric oxide (NO) release. Because the bradykinin/NO pathway may mediate some of the effects of angiotensin II AT2 receptors, we examined the possible contribution of AT2 receptors to the kinin dependent response to flow. Changes in outer diameter after increases in flow rate were evaluated in perfused arteries from wild-type animals (TK+/+) and in tissue kallikrein-deficient mice (TK-/-) in which the presence of AT2 receptor expression was verified. Saralasin, a nonselective angiotensin II receptor antagonist, impaired significantly flow-induced dilation in TK+/+, whereas it had no effect in TK-/- mice. In both groups, blockade of AT1 receptors with losartan or candesartan did not affect the response to flow. Inhibition of AT2 receptors with PD123319 reduced significantly flow-induced dilation in TK+/+ mice, but had no significant effect in TK-/- mice. Combining PD123319 with the bradykinin B2 receptor antagonist HOE-140 had no additional effect to AT2 receptor blockade alone in TK+/+ arteries. Flow-dependent-dilation was also impaired in AT2 receptor deficient mice (AT2-/-) when compared with wild-type littermates. Furthermore, HOE-140 significantly reduced the response to flow in the AT2+/+, but not in AT2-/- mice. In conclusion, this study demonstrates that the presence of functional AT2 receptors is necessary to observe the contribution of the vascular kinin-kallikrein system to flow-dependent dilation. PMID- 15131009 TI - Vascular endothelial growth factor receptor-2-induced initial endothelial cell migration depends on the presence of the urokinase receptor. AB - The angiogenic response of endothelial cells initiated by different growth factors is accompanied by assembly of cell surface-bound proteolytic machinery as a prerequisite for focal invasion. We have shown previously how the vascular endothelial growth factor (VEGF) initiates proteolysis by activation of pro urokinase (pro-PA) via the VEGF receptor-2 (VEGFR-2). We now show that the cell surface receptor of the uPA-system, the urokinase receptor (uPAR), is redistributed to focal adhesions at the leading edge of endothelial cells in response to VEGF. VEGF165 and VEGF-E, both interacting with VEGFR-2, but not PlGF exclusively stimulating VEGFR-1, induce within minutes internalization of uPAR via an LDL receptor-like molecule, dependent on generation of active uPA and the presence of plasminogen activator inhibitor-1 (PAI-1). uPAR seems to play a pivotal role in VEGFR-2-induced endothelial cell migration because cleavage of surface uPAR impaired the migratory response of endothelial cells toward VEGF-E, but not toward PlGF. PMID- 15131010 TI - Soy processing influences growth of estrogen-dependent breast cancer tumors. AB - Soy-based products consumed in Asian countries are minimally processed whereas in the USA many of the soy foods and soy ingredients are highly processed. Soy foods contain complex mixtures of bioactive compounds, which may interact with one another. The objective of this study was to evaluate the ability of various soy products containing genistin, the glycoside form of genistein, to affect growth of MCF-7 cells transplanted into ovariectomized athymic mice. Products investigated included soy flour, two crude extracts of soy (soy molasses and Novasoy(R)), a mixture of isoflavones and genistin in pure form. Each of the soy flour-processed products was added to the diet to provide equivalent amounts of genistein aglycone equivalents (750 p.p.m.). Tumors in the negative control animals regressed throughout the study while the tumors in the soy flour-fed animals remained basically the same size (neither grew nor regressed). In animals consuming soy molasses, Novasoy(R), mixed isoflavones or genistin alone, tumor growth was stimulated when compared with animals consuming a control diet devoid of soy. These same dietary treatments resulted in increased cellular proliferation. Changes in mRNA expression of gene targets (estrogen responsiveness, cell cycle progression, apoptosis and aromatase activity) in tumors induced by the different diets were evaluated. The relative expression of pS2, progesterone receptor and cyclin D1 was increased in animals consuming the Novasoy(R), mixed isoflavones and genistin. Bcl2 mRNA expression was low in most of the dietary treatment groups compared with positive (estradiol implant) controls. Aromatase expression was not affected in any of the treatment groups. The degree of soy flour processing affects the estrogenicity of products containing a constant amount of genistein. Collectively, these findings suggest that for postmenopausal women with estrogen-dependent breast cancer, the consumption of foods containing soy flour is more advisable than consuming isoflavones in more purified forms. PMID- 15131011 TI - Fibrates induce hepatic peroxisome and mitochondrial proliferation without overt evidence of cellular proliferation and oxidative stress in cynomolgus monkeys. AB - There is little primate risk factor data in the literature evaluating the relationship between proposed mechanisms of PPAR agonist-induced hepatocarcinogenesis at clinically relevant therapeutic exposures. These studies were conducted to characterize the hepatic effects of fenofibrate and ciprofibrate in the cynomolgus monkey. Male cynomolgus monkeys were given fenofibrate (250, 1250 or 2500 mg/kg/day) or ciprofibrate (3, 30, 150 or 400 mg/kg/day) for up to 15 days. The highest doses used were approximately 4 times (fenofibrate) and 9.4 times (ciprofibrate) the human therapeutic exposure for these agents based on AUC (area under the curve). For both compounds, there was a treatment-related increase in liver weight and periportal hepatocellular hypertrophy, which was related to increases in peroxisomes (up to 2.8 times controls) and mitochondria (up to 2.5 times controls). An increase in smooth endoplasmic reticulum probably contributed to the hypertrophy. There was no indication of cell proliferation as determined by the number of mitotic figures and this was confirmed by evaluating cell proliferation by immunohistochemical staining for the Ki-67 antigen. Consistent with the findings by light microscopy, there was no treatment-related effect on the level of mRNA for proteins known to be involved in the control of hepatocyte cell division or apoptosis (e.g. P21, Cyclin D1, PCNA, CDKN1A). Furthermore, there was minimal indication of oxidative stress. Thus, there was no evidence of lipofuscin accumulation, and there was no remarkable increase in the mRNA levels for most proteins known to respond to oxidative stress (e.g. catalase, glutathione peroxidase). A mild induction in the mRNA levels of cellular beta-oxidation and detoxification enzymes (e.g. acyl CoA oxidase, thioredoxin reductase) was observed. Collectively, the data from these studies suggest that the primate responds to PPARalpha agonists in a manner that is different from the rodent suggesting that the primate may be refractory to PPAR-induced hepatocarcinogenesis. PMID- 15131012 TI - Carcinogenesis Young Investigator Award. Telomere epigenetics: a higher-order control of telomere length in mammalian cells. AB - Telomeres are capping structures at the ends of eukaryotic chromosomes composed of TTAGGG repeats bound to an array of specialized proteins. Telomeres, together with centromeres, have been classically considered heterochromatic regions. Constitutive heterochromatin domains typically consist of repetitive DNA and have a very low gene content. In addition, constitutive heterochromatin is characterized by a number of hallmark histone modifications, as well as DNA modifications. In the case of pericentric heterochromatin, several activities responsible for these epigenetic modifications have been recently identified and characterized. In contrast, very little is still known on the architecture of telomeric chromatin, as well as on the activities that may regulate its structure and function. Here, we will discuss recent findings suggesting that telomeric chromatin shares many features with pericentric chromatin, and that disruption of telomeric heterochromatin results in changes in telomere length. PMID- 15131013 TI - Targeted overexpression of androgen receptor in osteoblasts: unexpected complex bone phenotype in growing animals. AB - The androgen receptor (AR), as a classic steroid receptor, generally mediates biologic responses to androgens. In bone tissue, both AR and the estrogen receptor (ER) are expressed in a variety of cell types. Because androgens can be converted into estrogen via aromatase activity, the specific role of the AR in maintenance of skeletal homoeostasis remains controversial. The goal of this study was to use skeletally targeted overexpression of AR as a means of elucidating the specific role(s) for AR transactivation in bone homeostasis. Rat AR cDNA was cloned downstream of a 3.6-kb alpha1(I)-collagen promoter fragment and used to create AR-transgenic mice. AR-transgenic males gain less weight and body and femur length is shorter than wild-type controls, whereas females are not different. AR-transgenic males also demonstrate thickened calvaria and increased periosteal but reduced endosteal labeling by fluorescent labeling and reduced osteocalcin levels. High-resolution micro-computed tomography shows normal mineral content in both male and female AR-transgenic mice, but male AR transgenics reveal a reduction in cortical area and moment of inertia. Male AR transgenics also demonstrate an altered trabecular morphology with bulging at the metaphysis. Histomorphometric analysis of trabecular bone parameters confirmed the increased bone volume comprised of more trabeculae that are closer together but not thicker. Biomechanical analysis of the skeletal phenotype demonstrate reduced stiffness, maximum load, post-yield deflection, and work-to-failure in male AR-transgenic mice. Steady-state levels of selected osteoblastic and osteoclastic genes are reduced in tibia from both male and female transgenics, with the exception of increased osteoprotegerin expression in male AR-transgenic mice. These results indicate that AR action is important in the development of a sexually dimorphic skeleton and argue for a direct role for androgen transactivation of AR in osteoblasts in modulating skeletal development and homeostasis. PMID- 15131014 TI - Glucocorticoids do not alter developmental expression of hippocampal or pituitary steroid receptor coactivator-1 and -2 in the late gestation fetal guinea pig. AB - Steroid receptor coactivator (SRC) proteins interact with glucocorticoid receptors in a ligand-dependent manner to enhance transcription. Although glucocorticoids are essential for normal brain maturation, little is known about the presence or regulation of SRC proteins in the developing central nervous system. In the current study we demonstrated that SRC-1 was highly expressed in the fetal limbic system (hippocampal CA3>CA1/2>CA4>dentate gyrus) at gestational d (gd) 40 (term, approximately 70 d), whereas SRC-2 was undetectable at all time points. Hippocampal SRC-1 mRNA and protein expression were reduced in male and female fetuses with advancing gestation. In contrast, SRC-1 mRNA levels increased significantly in the dentate gyrus near term. Repeated maternal injection (1 or 10 mg/kg on gd 40, 41, 50, 51, 60, and 61) with synthetic glucocorticoid had no effect on fetal limbic SRC-1 expression at gd 62 in either sex. SRC-1 and SRC-2 mRNA expression in the anterior pituitary did not change over the second half of gestation and was unaffected by prenatal exposure to synthetic glucocorticoid. In conclusion, SRC-1 expression undergoes spatial, temporal, and region-specific regulation during development, and limbic and pituitary SRC-1 and SRC-2 are not regulated by glucocorticoids in late gestation. Developmental changes in limbic SRC-1 expression probably have important consequences on steroid receptor signaling, which is known to be critical for brain maturation in late gestation. PMID- 15131016 TI - Evidence indicating that renal tubular metabolism of leptin is mediated by megalin but not by the leptin receptors. AB - Leptin is secreted by adipocytes and is a circulating factor that regulates food intake and energy expenditure. Its serum level is elevated in patients with renal failure and has been suggested to be associated with malnutritional factors in these patients. Leptin has been suggested to be primarily metabolized by the kidneys, although the precise molecular mechanisms are not known. The purpose of this study was to determine the nephron segments and potential receptors involved in renal leptin metabolism. To determine the segment involved in leptin uptake, we performed histoautoradiography of kidney sections obtained from rats that had been injected iv with (125)I-leptin. The ability of megalin, a multiligand endocytic receptor in the proximal tubules, to bind and endocytose leptin was examined by ligand blotting analysis, quartz-crystal microbalance, and degradation assays using megalin-expressing rat yolk sac L2 cells. Immunohistochemistry was performed to localize leptin receptors (LEP-R) in the rat kidney using two antibodies that recognize different epitopes on the LEP-R proteins. Circulating (125)I-leptin was filtered by glomeruli and internalized by proximal convoluted tubules. Megalin bound leptin in the presence of Ca(2+) and mediated its cellular internalization and degradation. On immunohistochemistry, LEP-R were localized in the proximal straight tubules, loops of Henle, distal tubules, and collecting ducts. In conclusion, circulating leptin was filtered by glomeruli and taken up by proximal convoluted tubules, where megalin likely mediates its binding and uptake. The localization of LEP-R suggests that they are not primarily involved in leptin metabolism in the proximal tubules. PMID- 15131015 TI - Immobilization induces acute nitric oxide production in the rat hypothalamus: a role of ionotropic glutamate receptors in the paraventricular nucleus. AB - Production of nitric oxide (NO) in the hypothalamic paraventricular nucleus (PVN) was examined by microdialysis in rats subjected to immobilization (IMO) stress. A dialysis probe was implanted in the posterior magnocellular subdivision of the PVN and nitrite (NO(2)(-)), an oxidized product of NO, was measured continuously. NO(2)(-) concentration in dialysate was enhanced to 156% after 30 min of IMO compared with the NO(2)(-) level before IMO. Intraperitoneal administration of N(G)-monomethyl-l-arginine (10 mg/kg), a NO synthase inhibitor, before IMO completely inhibited the increase of NO production that IMO was to induce. Depletion of catecholamines innervating the PVN by an injection of 6 hydroxydopamine into the lateral ventricle before the microdialysis had no suppressive effect on the increase of NO production by IMO. In contrast, NO(2)(-) levels in the PVN were lowered by continuous perfusion of the solution containing the ionotropic glutamate receptor antagonists 2-amino-5-phosphonovaleric acid (500 microm) and 6-cyano-7-nitroquinoxaline-2, 3 dione (50 microm) through the dialysis probe, and the IMO-induced increase of NO production was attenuated by the treatment. These results suggest that catecholaminergic drive to the hypothalamus is not necessary for the IMO-induced increase of NO production and that ionotropic glutamate receptors play a role in the basal and IMO-induced NO production. PMID- 15131017 TI - A membrane estrogen receptor mediates intracellular calcium release in astrocytes. AB - Appreciating the physiology of astrocytes and their role in brain functions requires an understanding of molecules that activate these cells. Estradiol may influence astrocyte functions. We now report that estrogen altered intracellular calcium concentration ([Ca(2+)](i)) in neonatal astrocytes that expressed estrogen receptor (ER) mRNA in vitro. Western blotting revealed both ERalpha and ERbeta proteins in both the nuclear fractions and plasma-membrane fractions. Application of 17beta-estradiol (20 nm) to fura 2-loaded astrocytes in vitro stimulated [Ca(2+)](i) in 75% of astrocytes with an EC(50) of 12.7 +/- 3.1 nm. This rapid action of estradiol was blocked by the ER antagonist, ICI 182,780. The membrane-impermeable estradiol-BSA induced a [Ca(2+)](i) flux that was statistically similar to estradiol. Removal of extracellular Ca(2+) did not alter the effect of estradiol, but phospholipase C inhibitor U73122 (10 microm) and 2 aminoethoxydiphenyl borate (5 microm), an inhibitor of the inositol-1,4,5, trisphosphate-gated intracellular Ca(2+) channel, significantly decreased the estradiol-induced [Ca(2+)](i) flux. Estradiol was unable to induce [Ca(2+)](i) flux in thapsigargin-depleted cells. These results indicate that estradiol mediates [Ca(2+)](i) flux in astrocytes through a membrane-associated ER that activates the phospholipase C pathway. PMID- 15131018 TI - Transcriptional activation of the insulin-like growth factor I receptor gene by the Kruppel-like factor 6 (KLF6) tumor suppressor protein: potential interactions between KLF6 and p53. AB - The IGF system plays an important role in prostate cancer initiation and progression. Most of the biological actions of IGF-I and IGF-II are mediated by activation of the IGF-I receptor (IGF-IR). Evidence accumulated in recent years indicates that acquisition of the malignant phenotype is initially IGF-IR dependent, but progression toward metastatic stages is usually associated with a decrease in IGF-IR levels. The Kruppel-like factor 6 (KLF6) is a zinc finger containing transcription factor that was shown to be mutated in a significant portion of prostate and other types of cancer. To examine the potential regulation of IGF-IR gene expression by KLF6, we measured KLF6 levels in prostate derived cell lines displaying different levels of IGF-IR. The results of Western analysis showed that KLF6 levels were higher in nontumorigenic P69 cells expressing high IGF-IR levels than in metastatic M12 cells containing reduced IGF IR levels. Transient coexpression of wild-type, but not mutated, KLF6 together with an IGF-IR promoter-luciferase reporter plasmid resulted in an approximately 3.4-fold stimulation of IGF-IR promoter activity. Furthermore, KLF6 expression induced a significant increment in endogenous IGF-IR levels. Deletion analysis of the IGF-IR promoter revealed that a cluster of four GC boxes located between nucleotides -399 and -331 mediates a significant portion of the transactivating effect of KLF6. KLF6, although unable to stimulate IGF-IR promoter activity in Sp1-null Drosophila-derived Schneider cells, significantly enhanced the effect of Sp1. To assess the potential interactions between KLF6 and p53 in the regulation of IGF-IR gene expression, transfections were performed in the colorectal cancer cell line HCT116(+/+), which expresses p53, and its HCT116(-/-) derivative, which lacks p53. KLF6 exhibited an enhanced activity in p53-containing, compared with p53-null, cells. In addition, we were able to detect a physical interaction between KLF6 and p53. In summary, we have identified the IGF-IR gene as a novel downstream target for transcription factor KLF6. The regulation of IGF-IR gene expression by KLF6 may have significant implications in terms of cancer initiation and/or progression. PMID- 15131019 TI - Identification and validation of novel androgen-regulated genes in prostate cancer. AB - Androgen-regulated genes (ARGs) are essential for the development of the prostate. Ironically, ARGs are also responsible for the pathogenesis of prostate cancer. We used oligonucleotide array technology to study the expression profiles of ARGs in LNCaP prostate cancer cells and identified 692 dihydrotestosterone regulated genes. Representative clusters containing genes with similar expression patterns to prostate-specific antigen and other known ARGs are discussed. Based on functional information, we categorized several candidate targets for prostate cancer therapy and diagnosis. Although many of these candidate targets are known to play an important role in cancer development, several are novel genes to the field of prostate cancer. A cross-comparison study of our results with those that have been previously published from three other array experiments using a similar LNCaP model validated 13 of these candidate targets as androgen-regulated. FKBP51 (FK506-binding immunophilin 51) was found in the same cluster as prostate specific antigen and its protein expression was increased in LNCaP cells treated with either dihydrotestosterone or synthetic androgen R1881. Results from mining the Gene Logic BioExpress database showed that FKBP51 expression is significantly higher in the prostate cancer group than in the normal and normal adjacent group. Additionally, the androgen-independent prostate tumor xenograft, CWR22R, had higher FKBP51 protein levels than that of the androgen-dependent prostate tumor xenograft, CWR22. A tissue microarray study further revealed that FKBP51 protein expression was higher in prostate cancer specimens than in benign prostate tumor samples. These results suggest the potential value of FKBP51 as a novel diagnostic marker or target for prostate cancer therapy. PMID- 15131020 TI - Peroxisome proliferator-activated receptor gamma agonism increases the capacity for sympathetically mediated thermogenesis in lean and ob/ob mice. AB - The nuclear receptor peroxisome proliferator-activated receptor (PPAR)gamma modulates the expression of numerous genes involved in glucose and lipid homeostasis and plays a critical role in adipocyte differentiation. Expression of uncoupling protein (UCP)1, which is necessary for thermogenesis, is strongly stimulated by PPARgamma agonists but without an increase in energy expenditure. This study was designed to assess whether PPARgamma-induced UCP1 has any functional impact and, if so, whether it involves sympathetic activity. In a first phase, obese ob/ob C57BL/6J mice and lean controls were treated for 2 wk with the PPARgamma agonist [2-(2-[4-phenoxy-2-propylphenoxy]ethyl)indole-5-acetic acid] (COOH). COOH induced UCP1 expression in brown and white adipose tissues as well as that of other genes associated with substrate oxidation and thermogenesis. However, UCP1 induction did not increase energy expenditure, as assessed by indirect calorimetry and other energy balance measurements. In a second phase, mice received for an additional 2 wk a combination of COOH and the beta(3)-adrenergic receptor (beta(3)-AR) agonist CL-316243 to stimulate the adrenergic signaling pathway and assess whether COOH-induced UCP1 was physiologically functional. The beta(3)-AR agonist stimulated thermogenesis in lean and ob/ob mice, an effect that was much stronger in COOH-pretreated mice, which exhibited lower respiratory quotient, higher oxygen consumption, and marked weight and fat mass loss, compared with mice not pretreated with COOH. These results demonstrate that PPARgamma agonism increases the thermogenic potential of white and brown adipose depots in lean and obese mice. This enhanced capacity leads to increased thermogenesis under beta-adrenergic stimulation, suggesting that the sympathetic drive is blunted by PPARgamma agonism. PMID- 15131021 TI - Denaturing HPLC-based approach for detecting RYR2 mutations involved in malignant arrhythmias. AB - BACKGROUND: Mutations in the RYR2 gene, which encodes the cardiac ryanodine receptor, have been reported in patients showing either arrhythmogenic right ventricular cardiomyopathy, type 2, or stress-induced polymorphic ventricular tachycardia. Both clinical phenotypes are characterized by a high risk of sudden death. Detection of RYR2 mutations is particularly important because beta-blocker treatment has been shown to be effective in preventing fatal arrhythmias in affected patients. METHODS: We used denaturing HPLC (DHPLC) to identify mutations in the human RYR2 gene. Fifty-three single exons, possibly targeted by mutations, were identified by comparison with the distribution of pathogenic mutations of the RYR1 gene, the skeletal muscle counterpart of RYR2. PCR primers for amplification of the entire coding sequence (116 amplicons, corresponding to 105 exons) were tested, and optimal DHPLC conditions were established. DHPLC analysis of critical exons was performed on 22 unrelated patients with effort-induced polymorphic ventricular arrhythmias but lacking a precise diagnosis. RESULTS: We identified four novel missense mutations among 22 patients. Their pathogenic role was related to present knowledge of the structure and function of RyR2 protein. CONCLUSIONS: Under optimized conditions, DHPLC is a cost-effective, highly sensitive, rapid, and efficient method for mutation screenings. A four-step approach is proposed for mutation screening of the RYR2 gene: (a) DHPLC analysis of 48 critical exons (2-4, 6-15, 17-20, 39-49, 83, 84, 87-97, and 99-105); (b) DNA sequencing of 5 critical exons unsuitable for DHPLC; then, in case of negative results, (c) DHPLC analysis of the remaining 39 exons and (d) DNA sequencing of the last 13 amplicons unsuitable for DHPLC analysis. PMID- 15131022 TI - Measurement of Nepsilon-(carboxymethyl)lysine and Nepsilon-(carboxyethyl)lysine in human plasma protein by stable-isotope-dilution tandem mass spectrometry. AB - BACKGROUND: N(epsilon)-(Carboxymethyl)lysine (CML) and N(epsilon) (carboxyethyl)lysine (CEL) are two stable, nonenzymatic chemical modifications of protein lysine residues resulting from glycation and oxidation reactions. We developed a tandem mass spectrometric method for their simultaneous measurement in hydrolysates of plasma proteins. METHODS: CML and CEL were liberated from plasma proteins by acid hydrolysis after addition of deuterated CML and CEL as internal standards. Chromatographic separation was performed by gradient-elution reversed-phase chromatography with a mobile phase containing 5 mmol/L nonafluoropentanoic acid as ion-pairing agent. Mass transitions of 205.1-->84.1 and 219.1-->84.1 for CML and CEL, respectively, and 209.1-->88.1 and 223.1-->88.1 for their respective internal standards were monitored in positive-ion mode. RESULTS: CML and CEL were separated with baseline resolution with a total analysis time of 21 min. The lower limit of quantification was 0.02 micromol/L for both compounds. Mean recoveries from plasma samples to which CML and CEL had been added were 92% for CML and 98% for CEL. Within-day CVs were <7.2% for CML and <8.2% for CEL, and between-day CVs were <8.5% for CML and <9.0% for CEL. In healthy individuals (n = 10), mean (SD) plasma concentrations of CML and CEL were 2.80 (0.40) micromol/L (range, 2.1-3.4 micromol/L) and 0.82 (0.21) micromol/L (range, 0.5-1.2 micromol/L), respectively. In hemodialysis (n = 17) and peritoneal dialysis (n = 9) patients, plasma concentrations of CML and CEL were increased two- to threefold compared with controls, without significant differences between dialysis modes [7.26 (1.36) vs 8.01 (3.80) micromol/L (P = 0.89) for CML, and 1.84 (0.39) vs 1.71 (0.42) micromol/L (P = 0.53) for CEL]. CONCLUSIONS: This stable-isotope-dilution tandem mass spectrometry method is suitable for simultaneous analysis of CML and CEL in hydrolysates of plasma proteins. Its robustness makes it suitable for assessing the value of these compounds as biomarkers of oxidative stress resulting from sugar and lipid oxidation. PMID- 15131023 TI - E1A as a tumor suppressor gene: commentary re S. Madhusudan et al. A multicenter Phase I gene therapy clinical trial involving intraperitoneal administration of E1A-lipid complex in patients with recurrent epithelial ovarian cancer overexpressing HER-2/neu oncogene. PMID- 15131024 TI - A Phase I and pharmacokinetic study of VNP40101M, a novel sulfonylhydrazine alkylating agent, in patients with refractory leukemia. AB - PURPOSE: VNP40101M is a novel sulfonylhydrazine alkylating agent with broad antitumor activity in animal models. As alkylating agents are important antileukemia drugs, a Phase I and pharmacokinetic study of VNP40101M was conducted in patients with refractory or relapsed leukemias or poor-risk myelodysplastic syndromes (MDS). EXPERIMENTAL DESIGN: VNP40101M was given as a single i.v. infusion over 15-70 min on day 1. Courses were repeated every 4 weeks according to antileukemic activity. The starting dose of 220 mg/m(2) was escalated by approximately 33% in cohorts of 3-6 patients until a maximum tolerated dose was established. One additional cohort was treated with the maximum-tolerated dose divided over days 1 and 8. RESULTS: Thirty-eight patients, including 28 with acute myeloid leukemia and 5 with MDS, received 52 courses of treatment. Nondose-limiting, reversible infusion-related toxicities were the most frequent adverse event, occurring in 24 (63%) patients on the first course. Dose escalation was terminated at 708 mg/m(2) for prolonged myelosuppression in 1 of 7 patients, and 600 mg/m(2) was selected as the recommended Phase II dose, with no significant extramedullary toxicity at this dose level. Two patients, 1 with MDS treated with 300 mg/m(2) and 1 with acute myeloid leukemia treated with 600 mg/m(2), achieved complete remission. CONCLUSIONS: VNP40101M had significant antileukemic activity and minimal extramedullary toxicity in patients with relapsed or refractory disease. PMID- 15131025 TI - BRCA mutations and risk of prostate cancer in Ashkenazi Jews. AB - PURPOSE: The Breast Cancer Linkage Consortium and other family-based ascertainments have suggested that male carriers of BRCA mutations are at increased risk of prostate cancer. Several series looking at the frequency of BRCA mutations in unselected patients with prostate cancer have not confirmed this finding. To clarify this issue, we conducted a large case-control study. EXPERIMENTAL DESIGN: Blood specimens from 251 unselected Ashkenazi men with prostate cancer were screened for the presence of one of the three common Ashkenazi founder mutations in BRCA1 and BRCA2. The incidence of founder mutations was compared with the incidence of founder mutations in 1472 male Ashkenazi volunteers without prostate cancer using logistic regression analysis after adjusting for age. RESULTS: Thirteen (5.2%) cases had a deleterious mutation in BRCA1 or BRCA2 compared with 28 (1.9%) controls. After adjusting for age, the presence of a BRCA1 or BRCA2 mutation was associated with the development of prostate cancer (odds ratio, 3.41; 95% confidence interval, 1.64 7.06; P = 0.001). When results were stratified by gene, BRCA2 mutation carriers demonstrated an increased risk of prostate cancer (odds ratio, 4.78; 95% confidence interval, 1.87-12.25; P = 0.001), whereas the risk in BRCA1 mutation carriers was not significantly increased. CONCLUSIONS: BRCA2 mutations are more likely to be found in unselected individuals with prostate cancer than age matched controls. These results support the hypothesis that deleterious mutations in BRCA2 are associated with an increased risk of prostate cancer. PMID- 15131026 TI - A cross-study comparison of gene expression studies for the molecular classification of lung cancer. AB - PURPOSE: Recent studies sought to refine lung cancer classification using gene expression microarrays. We evaluate the extent to which these studies agree and whether results can be integrated. EXPERIMENTAL DESIGN: We developed a practical analysis plan for cross-study comparison, validation, and integration of cancer molecular classification studies using public data. We evaluated genes for cross platform consistency of expression patterns, using integrative correlations, which quantify cross-study reproducibility without relying on direct assimilation of expression measurements across platforms. We then compared associations of gene expression levels to differential diagnosis of squamous cell carcinoma versus adenocarcinoma via reproducibility of the gene-specific t statistics and to survival via reproducibility of Cox coefficients. RESULTS: Integrative correlation analysis revealed a large proportion of genes in which the patterns agreed across studies more than would be expected by chance. Correlation of t statistics for diagnosis of squamous cell carcinoma versus adenocarcinoma is high (0.85) and increases (0.925) when using only the most consistent genes identified by integrative correlation. Correlations of Cox coefficients ranged from 0.13 to 0.31 (0.33-0.49 with genes selected for consistency). Although we find genes that are significant in multiple studies but show discordant effects, their number is approximately that expected by chance. We report genes that are reproducible by integrative analysis, significant in all studies, and concordant in effect. CONCLUSIONS: Cross-study comparison revealed significant, albeit incomplete, agreement of gene expression patterns related to lung cancer biology and identified genes that reproducibly predict outcomes. This analysis approach is broadly applicable to cross-study comparisons of gene expression profiling projects. PMID- 15131027 TI - DNA methylation profiles of lymphoid and hematopoietic malignancies. AB - PURPOSE: Aberrant methylation of the 5' gene promoter regions is an epigenetic phenomenon that is the major mechanism for silencing of tumor suppressor genes in many cancer types. The aims of our study were (a) to compare the methylation profiles of the major forms of hematological malignancies and (b) to determine the methylation profile of monoclonal gammopathy of undetermined significance (MGUS) and compare it with that of multiple myeloma (MM). EXPERIMENTAL DESIGN: We compared the aberrant promoter methylation profile of 14 known or suspected tumor suppressor genes in leukemias (n = 48), lymphomas (n = 42), and MMs (n = 40). We also examined the methylation profile of MGUS (n = 20), a premalignant plasma cell dyscrasia. The genes studied represent five of the six "hallmarks of cancer." RESULTS: Peripheral blood lymphocytes (n = 14) from healthy volunteers were negative for methylation of all genes, and methylation percentages in 41 nonmalignant tissues (peripheral blood mononuclear cells, bone marrows, and lymph nodes) from hematological patients were low (0-9%) for all 14 genes, confirming that methylation was tumor specific. Ten of the genes were methylated at frequencies of 29-68% in one or more tumor types, and the methylation indices (an indicator of overall methylation) varied from 0.25 to 0.34. With two exceptions, the methylation patterns of leukemias and lymphomas were similar. However, the pattern of MMs varied from the other tumor types for six genes. In general, the methylation pattern of MGUS was similar to that of MM, although the methylation frequencies were lower (the methylation index of MGUS was 0.15, and that of MM was 0.3). However, the methylation frequencies of six genes were significantly higher in MGUS than in control tissues. The relatively high frequencies of methylation in MGUS are consistent with it being a premalignant condition. CONCLUSIONS: The three major forms of lymphoid/hematopoietic malignancies show overlapping but individual patterns of methylation. PMID- 15131028 TI - Synergistic cytotoxicity and pharmacogenetics of gemcitabine and pemetrexed combination in pancreatic cancer cell lines. AB - PURPOSE: Gemcitabine is an inhibitor of ribonucleotide reductase (RR) and DNA synthesis and is an effective agent in the treatment of pancreas cancer. The present study investigates whether the multitargeted antifolate pemetrexed would be synergistic with gemcitabine against MIA PaCa-2, PANC-1, and Capan-1 pancreatic cancer cell lines. EXPERIMENTAL DESIGN: Cells were treated with gemcitabine and pemetrexed, and the type of drug interaction was assessed using the combination index. Cytotoxicity of gemcitabine was examined with inhibitors of (a) deoxycytidine kinase (dCK), which activates gemcitabine by phosphorylation, and (b) 5'-nucleotidase (drug dephosphorylation) and cytidine deaminase (drug deamination), the main inactivating enzymes. The effects of gemcitabine and pemetrexed on cell cycle were analyzed by flow cytometry, and apoptosis was examined by fluorescence microscopy. Finally, quantitative, real time PCR was used to study the pharmacogenetics of the drug combination. RESULTS: Synergistic cytotoxicity and enhancement of apoptosis was demonstrated, mostly with the sequence pemetrexed-->gemcitabine. Pemetrexed increased cells in S phase, the most sensitive to gemcitabine, and a positive correlation was found between the expression ratio of dCK:RR and gemcitabine sensitivity. Indeed, pemetrexed significantly enhanced dCK gene expression (+227.9, +86.0, and +135.5% in MIA PaCa-2, PANC-1, and Capan-1 cells, respectively), and the crucial role of this enzyme was confirmed by impairment of gemcitabine cytotoxicity after dCK saturation with 2'-deoxycytidine. CONCLUSIONS: These data demonstrate that the gemcitabine and pemetrexed combination displays schedule-dependent synergistic cytotoxic activity, favorably modulates cell cycle, induces apoptosis, and enhances dCK expression in pancreatic cancer cells. PMID- 15131029 TI - Dendritic cell vaccination in medullary thyroid carcinoma. AB - PURPOSE: Prognosis and treatment effectiveness for medullary thyroid carcinoma (MTC) are strictly related to tumor stage. Palliative treatment options show no significant benefit. A promising treatment approach for human cancer is based on the vaccination of autologous dendritic cells (DCs). EXPERIMENTAL DESIGN: The objective of this study was to evaluate the effectiveness of DC vaccines in MTC patients. Therefore, we generated autologous tumor lysate-pulsed DCs from 10 patients suffering from advanced MTC for repeated vaccination. Mature DCs were derived from peripheral blood monocytes by using CD14 magnetic bead selection and subsequent culture in the presence of granulocyte macrophage colony-stimulating factor, interleukin 4, and tumor necrosis factor alpha with or without addition of IFN-gamma. DCs were loaded with tumor lysate and further injected into a groin lymph node. Toxicity, tumor marker profile, immune response, and clinical response were determined. RESULTS: Vaccination was well tolerated and induced a positive immunological response in all of the tested patients as evaluated by in vivo delayed-type hypersensitivity reactivity or in vitro intracytoplasmic IFN gamma detection assay. Three patients had a partial response, 1 patient presented a minor response, and 2 patients showed stable disease. The remaining 4 patients had progressive disease. CONCLUSIONS: These data provide strong evidence that vaccination with tumor-lysate pulsed DCs results in the induction of a specific immune response in patients suffering from MTC. Objective clinical responses could be observed even for far-advanced disease. Therefore, we suggest that MTC is particularly suited for DC-based immunotherapy. PMID- 15131030 TI - Immunological responses in women with human papillomavirus type 16 (HPV-16) associated anogenital intraepithelial neoplasia induced by heterologous prime boost HPV-16 oncogene vaccination. AB - PURPOSE: The purpose is to study the immunogenicity of heterologous prime-boost human papillomavirus (HPV) oncogene vaccination in patients with anogenital intraepithelial neoplasia (AGIN). EXPERIMENTAL DESIGN: Twenty-nine women with high-grade AGIN received three i.m. doses of TA-CIN (HPV-16 L2/E6/E7 protein) at four weekly intervals followed by a single dermal scarification of vaccinia HPV 16/18 E6/E7 and were followed up for 12 weeks. Immunity to HPV-16 was assessed by lymphoproliferation, IFN-gamma enzyme-linked immunospot (ELISPOT), and ELISA. RESULTS: The patient group significantly responded to TA-CIN and not to the control antigen HPV-6 L2/E7 at all postvaccination time points when compared with baseline responses (P < or = 0.05). Ten of the patients showed at least a 3-fold increase in TA-CIN-specific proliferation at one or more time points after vaccination. Comparison of stimulation with HPV-16 E6- or E7-GST fusion proteins showed that proliferative responses were biased to HPV-16 E6. This bias was also seen by IFN-gamma ELISPOT using overlapping peptides, with HPV-16 E6- or E7 specific T cells being detected in 9 and 2 patients, respectively. In addition, vaccination resulted in the induction of antibodies against the HPV-16 oncoproteins. Of the 6 clinical responders, 2 patients showed both a proliferative TA-CIN-specific response and an E6-specific IFN-gamma response, whereas 3 other patients displayed E6-specific reactivity only. Stable disease was recorded in 19 patients, 8 of whom showed a concomitant TA-CIN-specific proliferative and/or E6-specific T-cell response. Of the 4 progressors, 2 failed to make a T-cell response and 2 responded by either proliferation or E6 ELISPOT alone. CONCLUSIONS: The prime-boost regimen is immunogenic in AGIN patients (humoral and cellular immunity), but there is no simple relationship between induction of systemic HPV-16-specific immunity and clinical outcome. Other factors that may play a role in the eradication of long-term established AGIN lesions need to be determined to identify the patient group that would benefit from immunotherapy with the vaccines used in this study. PMID- 15131031 TI - Intraperitoneal cisplatin with intraperitoneal gemcitabine in patients with epithelial ovarian cancer: results of a phase I/II Trial. AB - PURPOSE: The aims of this study were to determine the dose and schedule of i.p. cisplatin with i.p. gemcitabine in patients with persistent disease at second look assessment, the toxicity of this regimen, and the time to treatment failure and overall survival. EXPERIMENTAL DESIGN: We performed a Phase I/II evaluation of i.p. cisplatin at 75 mg/m(2) on day 1 with planned gemcitabine at 500, 750, 1000, or 1250 mg/m(2) i.p. on days 1, 8, and 15 on a 28-day schedule for four courses. Eligible patients completed surgical cytoreduction followed by adjuvant platinum-based chemotherapy. They had second-look assessment showing microscopic or macroscopic (< or =1 cm) disease, followed by i.p. port placement. RESULTS: The Phase I dose-limiting toxicity was grade 3 thrombocytopenia at day 15 on dose level 1 (n = 5). The protocol was amended, and the Phase II portion accrued to 30 patients, who were given i.p. cisplatin (75 mg/m(2)) on day 1 and gemcitabine at 500 mg/m(2) on days 1 and 8 on a 21-day schedule for four courses. Nine patients were removed from the study: one each for hypersensitivity, cellulitis, and i.p. port malfunction; two for progression of disease; and four for renal toxicity. Other toxicities included grade 3 nausea (7%) and transient grade 3 neuropathy (3%). Grade 1 or 2 neuropathy was frequently seen (80%). Five patients (17%) returned to the operating room at a median of 6 months (range, 1-20 months) after i.p. therapy for evaluation of abdominal pain; two patients had recurrence, and all had areas of fibrous tissue with encasement of the bowel. In two patients, the fibrous tissue was causing partial bowel obstruction. No other patients had symptoms prompting surgical exploration. Pharmacokinetic (PK) studies showed a median area under the curve (AUC) i.p. of 3041 h. micro M (range, 676-5702 h. micro M) and AUC in plasma of 4.0 h. micro M (range, 0.92-8.2 h. micro M) reached between 120 and 240 min; the pharmacological advantage was 759-fold (range, 217 1415-fold) for i.p. versus plasma drug levels. The mean residence time of gemcitabine with i.p. administration was 4.7 h. The median time to progression of the intent to treat population was 15.93 months (95% confidence interval, 9.13 25.9 months), with a median overall survival of 43.5 months [95% confidence interval, (34.66- infinity)]. No statistical differences were seen with respect to overall survival if patients were grouped in terms of optimal debulking or not (median not reached versus 34.8 months, respectively; P = 0.16) or whether visible disease was present or not at the start of i.p. therapy (34.8 versus 47.7 months; P = 0.47). With regard to time to treatment failure, a statistical difference favored patients with optimal versus nonoptimal primary debulking (25.2 versus 10.2 months, respectively; P = 0.03). CONCLUSIONS: The median time to treatment failure and overall survival of 15.9 months and 43.5 months, respectively, are consistent with our historical data in patients receiving i.p. platinum-based regimens for consolidation. The fibrotic changes seen in explored patients suggest local toxicity of this combination. The absolute benefit of i.p. consolidation requires randomized trials to assess efficacy. PMID- 15131032 TI - Phase I study of the farnesyltransferase inhibitor lonafarnib with paclitaxel in solid tumors. AB - PURPOSE: To establish the maximum tolerated dose of lonafarnib, a novel farnesyltransferase inhibitor, in combination with paclitaxel in patients with solid tumors and to characterize the safety, tolerability, dose-limiting toxicity, and pharmacokinetics of this combination regimen. EXPERIMENTAL DESIGN: In a Phase I trial, lonafarnib was administered p.o., twice daily (b.i.d.) on continuously scheduled doses of 100 mg, 125 mg, and 150 mg in combination with i.v. paclitaxel at doses of 135 mg/m(2) or 175 mg/m(2) administered over 3 h on day 8 of every 21-day cycle. Plasma paclitaxel and lonafarnib concentrations were collected at selected time points from each patient. RESULTS: Twenty-four patients were enrolled; 21 patients were evaluable. The principal grade 3/4 toxicity was diarrhea (5 of 21 patients), which was most likely due to lonafarnib. dose-limiting toxicities included grade 3 hyperbilirubinemia at dose level 3 (100 mg b.i.d. lonafarnib and 175 mg/m(2) paclitaxel); grade 4 diarrhea and grade 3 peripheral neuropathy at dose level 3A (125 mg b.i.d. lonafarnib and 175 mg/m(2) paclitaxel); and grade 4 neutropenia with fever and grade 4 diarrhea at level 4 (150 mg b.i.d. lonafarnib and 175 mg/m(2) paclitaxel). The maximum tolerated dose established by the continual reassessment method was lonafarnib 100 mg b.i.d. and paclitaxel 175 mg/m(2). Paclitaxel appeared to have no effect on the pharmacokinetics of lonafarnib. The median duration of therapy was eight cycles, including seven cycles with paclitaxel. Six of 15 previously treated patients had a durable partial response, including 3 patients who had previous taxane therapy. Notably, two of five patients with taxane-resistant metastatic non-small cell lung cancer had partial responses. CONCLUSIONS: When combined with paclitaxel, the recommended dose of lonafarnib for Phase II trials is 100 mg p.o. twice daily with 175 mg/m(2) of paclitaxel i.v. every 3 weeks. Additional studies of lonafarnib in combination regimens appear warranted, particularly in patients with non-small cell lung cancer. PMID- 15131033 TI - Plasma etoposide catechol increases in pediatric patients undergoing multiple-day chemotherapy with etoposide. AB - PURPOSE: The purpose of this research was to determine inter- and intrapatient differences in the pharmacokinetic profiles of etoposide and its genotoxic catechol metabolite during conventional multiple-day dosing of etoposide in pediatric patients. EXPERIMENTAL DESIGN: Seven pediatric patients with various malignancies received etoposide at a dose of 100 mg/m(2) i.v. over 1 h daily for 5 days. Blood samples were taken at selected time points on days 1 and 5. Plasma and protein-free plasma concentrations of etoposide and etoposide catechol were determined using a validated liquid chromatography/tandem mass spectrometry assay. Pharmacokinetic parameters of both etoposide and etoposide catechol were calculated using the WinSAAM modeling program developed at NIH. RESULTS: The mean maximum concentration (C(max)) for total (0.262 +/- 0.107 micro g/ml) and free catechol (0.0186 +/- 0.0082 micro g/ml) on day 5 were higher than the mean C(max) for total (0.114 +/- 0.028 micro g/ml) and free catechol (0.0120 +/- 0.0091 micro g/ml) on day 1. The mean area under the plasma concentration-time curve (AUC)(24h) for total (105.4 +/- 49.1 micro g.min/ml) and free catechol (4.89 +/- 2.23 micro g x min/ml) on day 5 were much greater (P < 0.05) than those for total (55.9 +/- 16.1 micro g x min/ml) and free catechol (3.04 +/- 1.04 micro g x min/ml) on day 1. In contrast, the AUC(24h) for etoposide was slightly lower on day 5 than on day 1. CONCLUSIONS: The C(max) and AUC(24h) for etoposide catechol were significantly higher on day 5 than on day 1. This suggests that metabolism of etoposide to its catechol metabolite increases in pediatric patients receiving multiple-day bolus etoposide infusions. These findings may be relevant to future reduction of the risk of leukemia as a treatment complication, because etoposide and etoposide catechol are both genotoxins. PMID- 15131034 TI - A multicenter Phase I gene therapy clinical trial involving intraperitoneal administration of E1A-lipid complex in patients with recurrent epithelial ovarian cancer overexpressing HER-2/neu oncogene. AB - PURPOSE: HER-2/neu oncogene is overexpressed in 10-30% of epithelial ovarian cancers and is associated with a poor prognosis. The E1A gene product of adenovirus type 5 down-regulates HER-2/neu and causes tumor regression in animal models. In the current study, we sought to determine the toxicity and biological activity of E1A-lipid complex in ovarian cancer patients. EXPERIMENTAL DESIGN: A Phase I trial involving intraperitoneal (i.p.) administration of E1A-lipid complex was initiated in ovarian cancer patients to assess biological activity (E1A gene transfer/transcription/translation and HER-2/neu expression) and to determine the maximum tolerated dose. Successive cohorts received E1A-lipid complex at doses of 1.8, 3.6, and 7.2 mg DNA/m(2), given as weekly i.p. infusions for 3 of 4 weeks (each cycle) up to a maximum of six cycles. Peritoneal fluid was sampled at baseline and twice monthly for cellularity, cytology, CA-125, and biological activity RESULTS: Fifteen patients, with a median age of 57 years (range, 43-81) were recruited. Three (1.8 mg DNA/m(2)), 4 (3.6 mg DNA/m(2)), and 8 patients (7.2 mg DNA/m(2)) received i.p. E1A. A total of 91 infusions (range, 1 18) was administered. Abdominal pain was the dose-limiting toxicity, and the maximum-tolerated dose was 3.6 mg DNA/m(2). E1A gene transfer and expression was observed in all of the patients and at all of the dose levels. HER-2/neu down regulation could be demonstrated in the tumor cells of 2 patients (18%). There was no correlation between dose and biological activity. CONCLUSIONS: I.P. EIA lipid complex gene therapy is feasible and safe. Future studies, either alone or in combination with chemotherapy, particularly in patients with minimal residual disease, should be evaluated. PMID- 15131035 TI - Somatostatin receptor subtype 2 is expressed by supratentorial primitive neuroectodermal tumors of childhood and can be targeted for somatostatin receptor imaging. AB - PURPOSE: Although gliomas predominate among central nervous system (CNS) neoplasms in adulthood, embryonal tumors are the most common malignant brain tumors in children. Despite novel treatment approaches, including improved radiotherapy and high-dose chemotherapy, survival rates remain unsatisfactory. The timely diagnosis of residual or recurrent embryonal CNS tumors and thus the earliest possible time point for intervention is often hampered by inaccuracies of conventional imaging techniques. Novel and refined imaging methodologies are urgently needed. EXPERIMENTAL DESIGN: We have previously demonstrated the use of somatostatin receptor imaging (SRI) in the diagnosis of recurrent and residual medulloblastomas. Here, we evaluated somatostatin receptor type 2 (sst(2)) expression using an antibody in an array of CNS tumors of childhood. Eight high grade gliomas, 4 atypical teratoid/rhabdoid tumors, 7 supratentorial primitive neuroectodermal tumors (stPNET), 1 medulloepithelioma (ME), and 8 ependymomas were screened. Tumors positive in vitro were additionally analyzed in vivo using SRI. RESULTS: Abundant expression of somatostatin receptor type 2 in stPNET, a ME, and ependymomas warranted in vivo imaging of 7 stPNET, 1 rhabdomyosarcoma, 3 ependymomas, 1 ME, and 1 glioblastoma. Although SRI was positive in 6/7 stPNET, 1 rhabdomyosarcoma, and 1 ME, none of the ependymomas nor the glioblastoma could be imaged using SRI. In selected cases SRI was more sensitive in the detection of relapse than conventional imaging by magnetic resonance imaging and computed tomography. CONCLUSIONS: SRI should be considered in the evaluation of residual or recurrent embryonal CNS tumors, especially stPNET. The strengths of SRI lie in the differentiation of reactive tissue changes versus residual or recurrent tumor, the detection of small lesions, and possibly in the distinction of stPNET from gliomas. PMID- 15131036 TI - Elevated expression of valosin-containing protein (p97) is associated with poor prognosis of prostate cancer. AB - PURPOSE: Valosin-containing protein (VCP) has been shown to be associated with metastasis and prognosis in human cancers. In the present study, the correlation of VCP with recurrence and prognosis in patients with prostate cancer (PCA) receiving conservative therapy was examined. EXPERIMENTAL DESIGN: VCP expression was analyzed immunohistochemically in 136 patients ranging from 46 to 92 years (median, 72 years), who received conservative therapy, including androgen deprivation, radiotherapy, or watchful waiting. Staining intensity of tumor cells was categorized as weaker (level 1) or equal to or stronger (level 2) than that in endothelial cells. The correlation of VCP expression between the mRNA and protein levels was examined in 10 patients. RESULTS: Thirty-two cases (23.5%) showed level 1 and 100 (76.5%) level 2 VCP expression. Quantitative reverse transcription-PCR analysis revealed greater VCPmRNA expression in level 2 (n = 5) than level 1 cases (n = 5; P < 0.05). A significant difference was observed between VCP level 1 and 2 patients in the positive rate for the digital rectal examination (P < 0.01), serum prostate-specific antigen level (P < 0.0001), cancer volume (P < 0.0001), Gleason score (P < 0.0001), stage (P < 0.0001), and progression-free and overall survival (P < 0.0001 for both). Multivariate analysis revealed VCP expression level, serum prostate-specific antigen level, and Gleason score to be independent prognosticators for progression-free and overall survival. Progression of PCA was found in 9.4% of level 1 but in 64% of level 2 patients. CONCLUSIONS: PCA with level 1 VCP expression could be treated conservatively. PMID- 15131037 TI - Comparative study between DNA copy number aberrations determined by quantitative microsatellite analysis and clinical outcome in patients with stomach cancer. AB - PURPOSE: We detected the relative DNA copy numbers (RCNs) at target loci in patients with stomach cancer with quantitative microsatellite analysis. We additionally clarified the relationship between DNA copy number aberrations and the clinical outcome of the patients. EXPERIMENTAL DESIGN: Fresh frozen samples were obtained from 30 patients who had undergone surgery for stomach cancer. Seven microsatellite loci in chromosomes 8q, 16q, and 20q and one gene-specific locus (ZNF217) were selected as the target loci. The DNA copy number was obtained relatively to a pooled reference consisting of six microsatellite primer sets selected from the regions where few aberrations have been reported in comparative genomic hybridization analysis. On the basis of the TaqMan PCR system, the internal probes used were carrying donor (6-carboxyfluorescein) and acceptor (6 carboxytetramethylrhodamine) fluorescent molecules complementary to CA repeats in the microsatellite markers and to one gene-specific oligomer in the gene-specific marker. RESULTS: Chromosome 8q gain, 20q gain, and 16q loss were detected in 18 (60.0%), 8 (26.7%), and 13 (43.3%) cases, respectively. Gains in the RCNs of D8S1801 and D8S1724 were most frequently found (36.7%). There was a significant correlation between the loss of D16S3026 and reduced survival duration (P = 0.0158), and the simultaneous aberrations of D8S1801 gain and D16S3026 loss (double marker positive) was significantly associated with reduced survival duration (P = 0.0008). According to Cox proportional hazards model, the double marker positive was a significant and independent factor indicating an unfavorable prognostic factor (relative risk, 17.176; 95% confidence interval, 2.782-106.026; P = 0.0022). CONCLUSION: RCN aberrations in tumor tissues determined by quantitative microsatellite analysis enable identification of the prognostic factors that correlate with clinical outcome of the patients with stomach cancer. PMID- 15131038 TI - Reliable and sensitive identification of occult tumor cells using the improved rare event imaging system. AB - PURPOSE: The purpose of this study was to assess the feasibility of using rare event imaging system (REIS)-assisted analysis to detect occult tumor cells (OTCs) in peripheral blood (PB). The study also sought to determine whether REIS assisted OTC detection presents a clinically viable alternative to manual microscopic detection to establish the true significance of OTC from solid epithelial tumors. EXPERIMENTAL DESIGN: We recently demonstrated proof of concept using a fluorescence-based automated microscope system, REIS, for OTC detection from the PB. For this study, the prototype of the system was adopted for high throughput and high-content cellular analysis. RESULTS: The performance of the improved REIS was examined using normal blood (n = 10), normal blood added to cancer cells (n = 20), and blood samples obtained from cancer patients (n = 80). Data from the screening of 80 clinical slides from breast and lung cancer patients, by manual microscopy and by the REIS, revealed that as many as 14 of 35 positive slides (40%) were missed by manual screening but positively identified by REIS. In addition, REIS-assisted scanning reliably and reproducibly quantified the total number of cells analyzed in the assay and categorized positive cells based on their marker expression profile. CONCLUSIONS: REIS-assisted analysis provides excellent sensitivity and reproducibility for OTC detection. This approach may enable an improved method for screening of PB samples and for obtaining novel information about disease staging and about risk evaluation in cancer patients. PMID- 15131039 TI - High level of glutathione-S-transferase pi expression in mantle cell lymphomas. AB - PURPOSE: Prognosis of mantle cell lymphoma (MCL) remains poor. Patients who achieve a response to first line therapy usually relapse, and the probability of cure remains low. Glutathione-S-transferase pi (GST-pi) overexpression has been associated with alkylating agents and anthracycline resistance. GST-pi gene is located in 11q13 and is coamplified along with CCND1 gene in some human solid tumors. EXPERIMENTAL DESIGN: We performed immunohistochemical analysis of GST-pi expression in 24 consecutive MCLs, 12 follicular lymphomas (FLs), and 69 diffuse large B-cell lymphomas (DLBCLs). Cases were classified in three groups: high GST pi expression (> 50% of cells were stained), moderate (5 to 50% cells were stained), or absent (< 5% cells were stained). GST-pi and CCND1 mRNA levels were also assessed by real-time reverse transcription-PCR analysis. RESULTS: All MCLs exhibit high GST-pi protein expression, compared with 29% of the DLBCLs and none of the FLs. MCLs expressed high levels of GST-pi and CCND1 mRNAs compared with DLBCLs and FLs. There was a strong relation between GST-pi and CCND1 mRNAs transcript levels in MCLs but not in DLBCLs. In conclusion, protein and mRNA GST pi expression is high in MCL compared with FL and DLBCL. CONCLUSIONS: Overexpression of CCND1 in MCL is associated with a transcriptional up-regulation of the GST-pi gene. Our results suggest that the glutathione system could play a role in drug resistance in MCL. PMID- 15131040 TI - Demethylation of urokinase promoter as a prognostic marker in patients with breast carcinoma. AB - PURPOSE: Urokinase (uPA) is expressed in a number of highly invasive malignancies including breast cancer. Because production of uPA is associated with breast cancer progression and can serve as a useful prognostic marker, the purpose of this study was to examine the role of uPA promoter methylation as an indicator of uPA production in breast cancer patients. EXPERIMENTAL DESIGN: We examined the methylation status of the uPA promoter and the levels of uPA expression in normal human breast epithelial cells and several human breast cancer cells by bisulfite sequencing analysis and reverse transcription-PCR. We also analyzed the methylation status of the uPA promoter in surgical biopsy samples from patients with breast cancer of different grades, as determined by the Elston-Ellis histological grading system. RESULTS: Expression of uPA mRNA was only detected in the highly invasive estrogen receptor-negative breast cancer cell lines, where the promoter was completely demethylated. In normal and low invasive breast cancer cells, the uPA promoter was methylated, resulting in lack of uPA mRNA expression. Analysis of biopsy samples showed that demethylation of the uPA promoter is associated with malignant transformation. Reverse transcription-PCR analysis revealed that this demethylation of the uPA promoter is directly associated with induction of uPA mRNA expression, which is well known to be associated with poor prognosis in breast cancer patients. CONCLUSIONS: This study indicated that uPA expression in breast cancer patients is under epigenetic control via methylation of its promoter. Determination of uPA promoter methylation can therefore serve as an early reliable indicator of uPA production in breast cancer patients. PMID- 15131041 TI - Intracellular patterns of Her-2/neu, ras, and ploidy abnormalities in primary human breast cancers predict postoperative clinical disease-free survival. AB - PURPOSE: In an earlier study, the presence of aneuploidy, Her-2/neu overexpression, and ras overexpression in the same cells (triple-positive cells) was of prognostic significance (P < 0.015) in 91 patients with localized breast cancer (median follow up, 32 months). Here, we present results involving a larger group of patients with longer follow-up. EXPERIMENTAL DESIGN: Fixed cell suspensions prepared from primary tumors of 189 patients with early breast cancer were studied prospectively by multiparameter flow cytometry. Correlated intracellular fluorescence-based measurements of cell DNA content and Her-2/neu and ras protein were obtained on each of >2000 cells in each tumor. Intracellular combinations of abnormalities in these measurements were correlated with subsequent patient disease-free survival (DFS). Median time on study was 54 months (range, 7-128 months). RESULTS: DFS of patients with > or = 5% triple positive tumor cells was shorter than those who did not meet this criterion (P = 0.004). The difference remained statistically significant after accounting for nodal status, tumor size, and each of the component abnormalities (P = 0.006). Node-negative patients whose tumors had fewer than 2 abnormalities/cell had an especially favorable clinical course, with a 5-year DFS of 96% (lower confidence bound, 86%). CONCLUSIONS: Patterns of accumulated intracellular molecular abnormalities in cells of primary human breast cancers are predictive for subsequent DFS independently of the abnormalities themselves taken individually. PMID- 15131042 TI - Loss of WWOX expression in gastric carcinoma. AB - PURPOSE: WW domain-containing oxidoreductase (WWOX) is a tumor suppressor gene that maps to the common fragile site FRA16D on chromosome 16q23.3-24.1. To investigate the role of the WWOX gene in the development of gastric carcinoma, we examined a large series of primary adenocarcinomas and nine gastric cancer cell lines for the expression of Wwox. EXPERIMENTAL DESIGN: Loss of heterozygosity, reverse-transcription-PCR, and immunohistochemistry were used to assess the role of WWOX in stomach cancer. A total of 81 primary gastric adenocarcinoma were analyzed. RESULTS: Loss of heterozygosity was observed in 31% of the cases and loss of Wwox protein expression was found in 65% of gastric adenocarcinoma primary specimens and 33% of gastric cancer cell lines. In addition, we found a high correlation between Wwox and Fhit protein expression. CONCLUSIONS: Our results indicate that alterations of the WWOX gene may be involved quite frequently in gastric tumorigenesis. Our data could be used in future studies to develop diagnostic and targeted therapy of stomach cancer. PMID- 15131043 TI - DNA aneuploidy and integration of human papillomavirus type 16 e6/e7 oncogenes in intraepithelial neoplasia and invasive squamous cell carcinoma of the cervix uteri. AB - PURPOSE: Increasingly deregulated expression of the E6-E7 oncogenes of high-risk human papillomaviruses (HR-HPVs) has been identified as the major transforming factor in the pathogenesis of cervical dysplasia and derived cancers. The expression of these genes in epithelial stem cells first results in chromosomal instability and induces chromosomal aneuploidy. It is speculated that this subsequently favors integration of HR-HPV genomes into cellular chromosomes. This in turn leads to expression of viral cellular fusion transcripts and further enhanced expression of the E6-E7 oncoproteins. Chromosomal instability and aneuploidization thus seems to precede and favor integration of HR-HPV genomes. EXPERIMENTAL DESIGN: To prove this sequential concept, we analyzed here the sequence of events of DNA aneuploidization and integration in a series of HPV-16 positive cervical dysplastic lesions and carcinomas. Eighty-five punch biopsies of HPV-16-positive cervical lesions (20 CIN1/2, 50 CIN3, and 15 CxCa) were analyzed for DNA ploidy by DNA flow cytometry and for integration of HPV E6/E7 oncogenes using the amplification of papillomavirus oncogene transcripts assay, a reverse transcription-PCR method to detect integrate-derived human papillomavirus oncogene transcripts. RESULTS: DNA aneuploidy and viral genome integration were both associated with increasing dysplasia (P < 0.001, chi(2) test for trend). In addition, DNA aneuploidy was associated with increased viral integration (P < 0.01, Fisher's exact test). Nineteen of 20 (95%) lesions with integrated viral genomes had aneuploid cell lines; however, only 19 of 32 (59%) lesions with aneuploid cell lines had integrated viral genomes. CONCLUSIONS: These data support the hypothesis that aneuploidization precedes integration of HR-HPV genomes in the progression of cervical dysplasia. Accordingly, deregulated viral oncogene expression appears to result first in chromosomal instability and aneuploidization and is subsequently followed by integration of HR-HPV genomes in the affected cell clones. PMID- 15131044 TI - Loss of 14-3-3sigma in prostate cancer and its precursors. AB - PURPOSE: The 14-3-3 family proteins are highly conserved over many mammalian species. The sigma isoform (also called HME-1 or stratifin) is expressed in epithelial cells. Loss of 14-3-3sigma is associated with failure to arrest the cell cycle at the G(2)-M phase checkpoint after DNA damage that leads to increased G(2)-type chromosomal aberrations. The role of 14-3-3sigma in prostatic carcinogenesis is uncertain. EXPERIMENTAL DESIGN: We studied one hundred and eleven specimens of invasive prostate adenocarcinoma with paired, adjacent high grade prostatic intraepithelial neoplasia and normal prostate epithelium. Immunohistochemistry was used to detect the expression of 14-3-3sigma. The findings were correlated with various clinical pathological parameters. RESULTS: 14-3-3sigma is ubiquitously expressed at high levels in normal prostate epithelium. Its expression is significantly decreased in prostatic intraepithelial neoplasia and prostatic adenocarcinoma. Ninety percent of samples of prostatic intraepithelial neoplasia had no or low 14-3-3sigma expression. Ninety-seven percent of invasive adenocarcinomas had no or low 14-3-3sigma expression. In most specimens (90%), suppression of 14-3-3sigma expression occurred during the development of prostatic intraepithelial neoplasia from normal epithelium. CONCLUSIONS: Our data suggest that loss of 14-3-3sigma contributes to the development of prostate adenocarcinoma. 14-3-3sigma expression is significantly decreased during the progression of normal prostatic epithelium to prostatic intraepithelial neoplasia and invasive cancer. PMID- 15131045 TI - Epidermal growth factor receptor, c-MET, beta-catenin, and p53 expression as prognostic indicators in stage II colon cancer: a tissue microarray study. AB - PURPOSE: Through the use of molecular markers, it may be possible to identify aggressive tumor phenotypes and tailor therapies to treat them. This approach would be particularly useful for stage II colon cancer. The purpose of this study was to define the prognostic value of epidermal growth factor receptor (EGFR), c MET, beta-catenin, and p53 protein expression in TNM stage II colon cancer using tissue microarray technology. EXPERIMENTAL DESIGN: In this study, we retrospectively analyzed, resected, and otherwise untreated paraffin-embedded specimens from 134 consecutive patients with Tumor-Node-Metastasis stage II colonic carcinomas for EGFR, c-MET, beta-catenin, and p53 protein expression by immunohistochemistry. RESULTS: Thirty-five percent, 77, and 65% of tumors exhibited strong (+2 and +3 immunopositivity) expression of EGFR, c-MET, and beta catenin, respectively. Fifty-four percent exhibited nuclear staining for p53 in >10% of the tumor cells. Univariate analysis revealed that increased nuclear p53 expression (P = 0.001), strong membranous EGFR expression (P = 0.04), and lymphovascular invasion (P = 0.01) were significantly related to disease recurrence and that p53 (P = 0.02) and EGFR (P = 0.05) expression were associated with decreased survival. Increased nuclear p53 expression also correlated with the presence of distal metastasis (P = 0.027). No significant association was seen between c-MET expression and prognosis, whereas a strong trend was detected between loss of beta-catenin (P = 0.065) expression and poor outcome. Multivariate analysis indicated that p53 (P = 0.04), EGFR (P = 0.05), and lymphovascular invasion (P = 0.03) were independent predictors of recurrence and that p53 (P = 0.02) and EGFR (P = 0.01) expression were both associated with poor survival. CONCLUSIONS: This retrospective tumor microarray study, restricted to Tumor-Node-Metastasis stage II colon cancer patients who did not undergo adjuvant therapy, supports the use of EGFR and p53 as biological markers, which may assist in predicting disease recurrence and survival. PMID- 15131046 TI - Prognostic significance of c-kit mutation in localized gastrointestinal stromal tumors. AB - PURPOSE: Constitutive mutational activation of c-kit has been found to be associated with the pathogenesis of gastrointestinal stromal tumors (GISTs). The prognostic significance of c-kit mutations, however, is still controversial. EXPERIMENTAL DESIGN: We examined 86 patients curatively resected for localized GIST. Genomic DNA was extracted from paraffin-embedded tumor tissues. Exons 9, 11, 13, and 17 of the c-kit gene were amplified by PCR and sequenced. RESULTS: Mutations in exon 11 were detected in 61 tumors, and mutations in exon 9 were observed in three tumors, whereas no mutations were detected in exons 13 or 17. The overall c-kit mutation frequency was 74%. Amino acid alterations in the 61 tumors with exon 11 mutations were deletion in 33 tumors, substitution in 20, both deletion and substitution in 4, insertion in 1, and duplication in 3. Histologically, tumors with c-kit mutations showed higher mitotic counts and higher cellularity. The 5-year relapse-free survival (RFS) in patients having GISTs with c-kit mutations was 21%, compared with 60% in those without c-kit mutations. Significantly higher RFS rates were observed in patients with tumors having mitotic counts < 5 mitoses/50 high power field, spindle-cell histology, tumor size < 5 cm, or gastric GISTs. Multivariate analyses indicated association of poorer RFS with a higher mitotic count > or = 5 of 50 high power fields; odds ratio (OR) = 3.0], presence of c-kit mutations (OR = 5.6), and a larger tumor size (> or = 5 cm; OR = 4.2). CONCLUSIONS: The presence of c-kit mutation, along with high mitotic count and larger tumor size, was an independent factor for poor prognosis in patients with localized GISTs. PMID- 15131047 TI - Molecular differences between sporadic serrated and conventional colorectal adenomas. AB - PURPOSE: The purpose is to compare the molecular characteristics of serrated adenomas (SAs) with those of conventional adenomas (CADs) and hyperplastic polyps (HPs). EXPERIMENTAL DESIGN: We evaluated the proliferative activity and molecular alterations in 47 SAs (25 pure-type and 22 mixed-type), 71 CADs, and 23 HPs. RESULTS: The proliferative activity of SAs, as evaluated by Ki-67 expression, was intermediate between CADs and HPs. There was no significant difference in the incidence of KRAS or p53 mutations between the three histological groups. In the microsatellite instability (MSI) analysis, 21% of SAs (9 of 43) showed MSI at two or more loci (MSI-H); corresponding values were 5% of CADs (3 of 64) and 8% of HPs (1 of 13; SAs versus CADs, P = 0.0125). MSI-H was more likely to be found in pure-type SAs (36%; 8 of 22) than in mixed-type SAs (5%; 1 of 21; P = 0.0212). Loss of hMLH-1 expression was found in 8 of 9 SAs with MSI-H. The incidence of BRAF or KRAS mutations was 36 and 15% of SAs, respectively; the combined incidence of BRAF and KRAS mutations occurred in 49% of SAs. However, there was no significant difference in the incidence of BRAF or KRAS mutations between SAs with and without MSI-H. CONCLUSIONS: Genetic instability is more frequently implicated in the tumorigenesis of SAs, especially pure-type SAs, than in that of CADs. In contrast, activation of the Ras/Raf/MEK/MAP kinase cascade by BRAF or KRAS mutation, independently of the genetic instability, may be associated with the progression of about half of SAs. PMID- 15131048 TI - The prognostic value of isolated tumor cells in bone marrow in breast cancer patients: evaluation of morphological categories and the number of clinically significant cells. AB - PURPOSE/EXPERIMENTAL DESIGN: Immunocytochemical detection of isolated tumor cells (ITCs) in the bone marrow (BM) is a prognostic factor in breast cancer. However, hematopoietic cells (HCs) can occasionally be stained by the techniques used. Morphological evaluation improves the specificity of ITC detection, but optimal separation of ITCs from false-positive HCs needs to be determined. Here, predetermined morphological categories of immunocytochemically (ICC)-positive cells in the BM and the number of detected ITCs were analyzed for association with clinical outcome in 817 early-stage breast cancer patients (median 49 months of follow-up). All ICC+ cells detected were categorized into one of the following groups: (a) tumor cell (TC); (b) uninterpretable cell (UIC); (c) probable HC; or (d) HC. RESULTS: Among the TC+ patients, 30.6% and 25.9% experienced systemic relapse (SR) and breast cancer death (BCD), respectively, as compared with 13.3% and 8.5% of patients without TCs in the BM (survival analyses: P < 0.001, log rank). The SR and BCD rate was 19.7% and 15.8% for TC-/UIC+ patients versus 12.5% and 7.4% for TC-/UIC- patients. Survival analyses confirmed that the UIC+ group contained clinically significant cells (P = 0.018, log-rank). No difference in clinical outcome was observed, regardless of whether probable HCs or HCs were present. Analyzing the number of ITC+ cells, SR and BCD occurred in 12.4% and 7.4% of patients with 0 ITCs present, 21.3% and 18.5% of patients with 1 ITC present, 19.4% and 16.7% of patients with 2 ITCs present, and 42.5% and 32.5% of patients with > or = 3 ITCs present. CONCLUSIONS: Morphological categorization of ICC+ cells improves the clinical value of ITC detection in the BM. The presence of only one ITC reduces survival, and a greater number of ITCs further aggravates the prognosis. PMID- 15131049 TI - Frequent loss of pRb2/p130 in human ovarian carcinoma. AB - PURPOSE: RB2/p130, a member of the retinoblastoma gene family, maps to human chromosome 16q12.2, a region in which deletions have been found in several human neoplasms including breast, prostatic, and ovarian carcinoma. We sought to evaluate pRb2/p130 protein expression and function in ovarian carcinoma. EXPERIMENTAL DESIGN: pRb2/p130 expression was detected by immunohistochemical and Western blot analyses in 45 primary ovarian carcinoma samples. RESULTS: Immunohistochemical analysis revealed loss or decrease of pRb2/p130 expression in 18 cases (40%). pRb2/p130 expression was mostly nuclear and inversely correlated to the tumor grade (P < 0.05). Western blot analysis correlated with immunohistochemical expression. Reverse transcription-PCR followed by Southern blot analysis was performed on a representative set of 20 ovarian carcinomas. RB2/p130 mRNA levels were consistent with protein expression. We found a significant increase in the percentage of G(1)-phase-arrested cells in CAOV3 and A2780 ovarian carcinoma cell lines after transduction with an adenovirus carrying the RB2/p130 gene (Ad-CMV-RB2/p130). CONCLUSIONS: These data indicate that loss or decrease of pRb2/p130 expression is a frequent event in ovarian carcinoma and is regulated mostly at the transcriptional level. Moreover, pRb2/p130 overexpression is able to arrest cell growth in ovarian carcinoma cells, suggesting the putative role of pRb2/p130 as a tumor suppressor in this malignancy. PMID- 15131050 TI - Very high frequency of hypermethylated genes in breast cancer metastasis to the bone, brain, and lung. AB - PURPOSE: Most often it is not the primary tumor, but metastasis to distant organs that results in the death of breast cancer patients. To characterize molecular alterations in breast cancer metastasis, we investigated the frequency of hypermethylation of five genes (Cyclin D2, RAR-beta, Twist, RASSF1A, and HIN-1) in metastasis to four common sites: lymph node, bone, brain, and lung. EXPERIMENTAL DESIGN: Methylation-specific PCR for the five genes was performed on DNA extracted from archival paraffin-embedded specimens of paired primary breast cancer and its lymph nodes (LN) metastasis (n = 25 each); in independent samples of metastasis to the bone (n = 12), brain (n = 8), and lung (n = 10); and in normal bone, brain, and lung (n = 22). RESULTS: No hypermethylation was detected in the five genes in the normal host tissues. In paired samples, LN metastasis had a trend of higher prevalence of methylation compared with the primary breast carcinoma for all five genes with significance for HIN-1 (P = 0.04). Compared with the primary breast carcinomas, all five genes had higher methylation frequencies in the bone, brain, and lung metastasis, with HIN-1 and RAR-beta methylation being significantly higher (P < 0.01) in each group. Loss of expression of all five genes correlated, with a few exceptions, to hypermethylation of their promoter sequences in metastatic carcinoma cells microdissected from LNs. CONCLUSION: The frequent presence of hypermethylated genes in locoregional and distant metastasis could render them particularly susceptible to therapy targeted toward gene reactivation combining demethylating agents, histone deacetylase inhibitors, and/or differentiating agents. PMID- 15131051 TI - Expression of matrix metalloproteinase-9 in head and neck squamous cell carcinoma: a potential marker for prognosis. AB - PURPOSE: Previous studies have shown that matrix metalloproteinase-9 (MMP-9) is expressed in malignant head and neck squamous cell carcinoma. The prognostic role of MMP-9 is still unclear. The aim of this study was to investigate the role of MMP-9 immunoreactive protein as a prognostic marker for survival in head and neck squamous cell carcinoma. EXPERIMENTAL DESIGN: Overexpression of the immunoreactive protein for MMP-9 was evaluated in tissue sections of 74 primary head and neck carcinomas with a monoclonal antibody using a biotin-streptavidin immunohistochemical staining method. The staining results were compared with the clinical data and to the patients' outcome. RESULTS: Positive immunostaining for MMP-9 was observed in 82% of the head and neck carcinomas, 39% of the cases being extensively positive. MMP-9 protein expression was independent of the stage or the grade of the tumor. The expression of MMP-9 was prognostic for shortened survival, the 5-year cause-specific survival being 45% in MMP-9 positive cases, and 92% in cases negative for MMP-9 (P = 0.013). MMP-9 positivity also correlated to the relapse-free survival (P = 0.019). At the 5-year follow-up, the cumulative relapse-free survival rate was 79% for patients with MMP-9-negative tumor and 42% for the patients with positive immunostaining for MMP-9. High expression of MMP-9 seemed to be linked with more aggressive relapses, appearing in 33% of the cases in local relapses, in 52% of cases with lymph node relapses, and in 60% of the cases with hematogenic relapses. CONCLUSIONS: This is the first study with a long follow-up showing that the immunoreactive protein of MMP-9 in head and neck carcinoma is associated with shortened relapse-free and cause-specific survival, suggesting that MMP-9 has a role in tumor progression of head and neck carcinomas, as well as in estimation of the prognosis of these diseases. PMID- 15131052 TI - Prognostic role of the ratio between cyclooxygenase-2 in tumor and stroma compartments in cervical cancer. AB - PURPOSE: The aim of this study was to analyze the clinical role of cyclooxygenase (COX)-2 in a large series of 175 cervical cancer patients. EXPERIMENTAL DESIGN: Immunohistochemistry was performed on paraffin-embedded sections by using rabbit antiserum against COX-2. The tumor:stroma (T/S) ratio of COX-2 expression was used to define the overall COX-2 content in the tumor. RESULTS: The T/S COX-2 ratio values ranged from 0.03 to 48.2 (mean +/- SE, 3.7 +/- 0.5). A total of 95 of 175 patients (54.3%) were scored as having a high (>1) T/S COX-2 ratio. In locally advanced cervical cancer patients who underwent neoadjuvant treatment, the percentage of cases showing a high T/S COX-2 ratio was greater in patients who did not respond to treatment (26 of 29 patients, 89.7%) than in patients with a partial (32 of 50 patients, 64.0%) or complete (19 of 44 patients, 43.2%) response (P = 0.0003). When logistic regression was applied, International Federation of Gynecologists and Obstetricians (FIGO) stage (chi(2) = 11.3; P = 0.0008) and T/S COX-2 ratio (chi(2) = 5.3; P = 0.021) retained an independent role in predicting a poor chance of response. Cases with a high T/S COX-2 ratio had a shorter overall survival (OS) [2-year OS, 61%(95% confidence interval 750 83)] than cases with a low T/S COX-2 ratio (2-year OS, 90%; 95% confidence interval, 81-99; P = 0.0001). In multivariate analysis, the status of T/S COX-2 IDV ratio, together with advanced stage, retained an independent negative prognostic role for OS. CONCLUSIONS: The assessment of COX-2 status in both tumor and stroma compartment could provide valuable information to identify cervical cancer patients endowed with a very poor chance of response to neoadjuvant treatment and unfavorable prognosis. PMID- 15131053 TI - Death-associated protein kinase loss of expression is a new marker for breast cancer prognosis. AB - PURPOSE: Death-associated protein (DAP)-kinase is a new Ser/Thr kinase involved in cell apoptosis and tumor suppression, the expression of which has been correlated to invasive potential and metastasis in several human neoplastic tissues. We analyzed the level of DAP-kinase expression in breast cancer specimens and its correlation with survival. EXPERIMENTAL DESIGN: One hundred twenty-eight breast cancer specimens were analyzed by immunohistochemistry. Patient records were studied retrospectively for demographic characteristics, clinical data, hormonal treatment, outcome, and survival. DAP-kinase protein expression was also studied in normal breast cells primary cultures under estrogen and antiestrogen treatment. RESULTS: Among the 128 patients, 30 showed a DAP-kinase staining < or = 20%, whereas 98 had a staining over 20%. Mean follow up time was 62 months. The association between tumor Scarff-Bloom and Richardson grade (P = 0.009), estrogen receptor and progesterone receptor expression (P = 0.002 and 0.001, respectively), tumor size (P = 0.05), Bcl-2 expression (P = 0.004), and DAP-kinase immunostaining in the ductal carcinoma group was highly significant. Overall (64 months) and disease-free (63 months) survival in the high DAP-kinase expression group were significantly longer compared with the women whose tumors showed a loss of DAP-kinase expression (51 and 43 months, respectively). DAP-kinase protein was strongly expressed in normal breast tissue and in human breast epithelial cells primary cultures. Estradiol decreased DAP kinase expression in these cells, arguing for hormonal regulation of the protein. CONCLUSIONS: Loss of DAP-kinase expression negatively correlates to survival and positively correlates to the probability of recurrence in a very significant manner. DAP-kinase thus constitutes a novel and independent prognosis marker for breast cancer. PMID- 15131054 TI - Overexpression of epithelial cell adhesion molecule antigen in gallbladder carcinoma is an independent marker for poor survival. AB - PURPOSE: Gallbladder carcinoma is an aggressive type of cancer that is difficult to cure by conventional procedures. There thus is a need to identify novel molecular markers for the assessment of prognosis and as potential therapeutic targets. This retrospective study was designed to investigate the prognostic significance of epithelial cell adhesion molecule (Ep-CAM) overexpression in human gallbladder carcinoma. EXPERIMENTAL DESIGN: Ep-CAM expression was examined immunohistochemically on paraffin-embedded tissue specimens from 99 patients who underwent surgical treatment for gallbladder carcinoma in the period between August 1988 and May 1999. RESULTS: Ep-CAM overexpression was found in 63 (63.6%) of the tumor samples. Kaplan-Meier curves showed that Ep-CAM overexpression was significantly related to decreased overall survival (P < 0.01). Overall survival gradually worsened with increasing Ep-CAM scores. Notably, in the subgroup of pT1 tumors (n = 17), patients without Ep-CAM overexpression had a 5-year overall survival rate of 100% compared with 38% (P = 0.01) for patients with Ep-CAM overexpressing tumors. By univariate analysis, no correlation was found with conventional clinicopathological parameters. Multivariate analysis, including Ep CAM expression, pT stage, tumor grade, and resection margin involvement, showed that Ep-CAM overexpression was an independent prognostic marker in gallbladder carcinoma (P = 0.03; relative risk, 1.8). CONCLUSIONS: These results demonstrate for the first time that Ep-CAM overexpression is an independent prognostic marker in gallbladder carcinoma and that its prognostic impact should be validated prospectively. Furthermore, the Ep-CAM antigen represents an attractive target for specific therapies with monoclonal antibodies or specific vaccines in patients with Ep-CAM-overexpressing gallbladder carcinoma. PMID- 15131055 TI - Pretargeted alpha emitting radioimmunotherapy using (213)Bi 1,4,7,10 tetraazacyclododecane-N,N',N",N"'-tetraacetic acid-biotin. AB - PURPOSE: The use of an alpha emitter for radioimmunotherapy has potential advantages compared with beta emitters. When administered systemically optimal targeting of intact antibodies requires >24 h, therefore limiting the use of short-lived alpha emitters. This study investigated the biodistribution of bismuth-labeled biotin in A431 tumor-bearing mice pretargeted with antibody B3 streptavidin (B3-SA) and examined the therapeutic efficacy of the alpha emitter, (213)Bi-labeled biotin. EXPERIMENTAL DESIGN: Biotinidase-resistant 7,10 tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA)-biotin was radiolabeled with (205,206)Bi or (213)Bi. Treatment of tumor-bearing mice began by administration of B3-SA (400 micro g) to target the tumor sites for 24 h. Then, an agent containing biotin and galactose groups was used to clear the conjugate from the circulation. Four h later, bismuth-radiolabeled DOTA-biotin was given, and biodistribution or therapy was evaluated. Dose escalation treatment from 3.7 74 MBq was performed, and the effects on tumors of different sizes were investigated. Tumor growth, complete blood cell counts, toxicity, and survival were monitored. RESULTS: Radiolabeled biotin cleared rapidly. Rapid tumor uptake resulted in much higher tumor:nontumor targeting ratios than achieved with the directly labeled monoclonal antibody. Dose escalation revealed that 74 MBq caused acute death of mice, whereas 0.37-37 MBq doses inhibited tumor growth and prolonged survival significantly. Evidence of mild hematological toxicity was noted. At therapeutically effective doses renal toxicity was observed. CONCLUSIONS: (213)Bi-DOTA-biotin, directed by the Pretarget method to tumor targeted B3-SA, showed a therapeutic effect, although the therapeutic index was low. The source of the toxicity was most likely related to the renal toxicity. PMID- 15131056 TI - Development of a new isogenic cell-xenograft system for evaluation of NAD(P)H:quinone oxidoreductase-directed antitumor quinones: evaluation of the activity of RH1. AB - PURPOSE: The purpose of our study was to develop and validate an isogenic cell line pair that differs only in the expression of NAD(P)H:quinone oxidoreductase (NQO1) that can be used to examine the in vitro and in vivo role of NQO1 in the bioactivation of the antitumor quinone RH1 (2,5-diaziridinyl-3-(hydroxymethyl)-6 methyl-1,4-benzoquinone), a compound currently in Phase I clinical trials. EXPERIMENTAL DESIGN: MDA-MB-468 (MDA468) human breast adenocarcinoma cells, homozygous for a polymorphism in NQO1 (NQO1*2/*2) and with low levels of NQO1 activity, were stably transfected with human NQO1 to generate a clone (NQ16) expressing very high NQO1 activity. We examined levels of other reductases and looked at biochemical systems that might influence response to antitumor quinones to validate that the isogenic cell line pair differed only in the expression of NQO1. The 3-(4,5-dimethylthiazol-2,5-diphenyl)tetrazolium (MTT) assay was used to determine the differential toxicity of various quinones, including the most recent NQO1-directed antitumor quinone, RH1, between the two cell lines. Human tumor xenografts were established from both MDA468 and NQ16 cells, and the antitumor activity of RH1 was evaluated. RESULTS: Levels of cytochrome P450 reductase, cytochrome b(5) reductase, soluble thiols, and superoxide dismutase in the NQ16 line were unchanged from the parental line. The functional significance of wild-type NQO1 expression was confirmed by measurement of the differential toxicity of compounds activated or deactivated by NQO1 in the two cell lines. The toxicity of the NQO1-directed antitumor quinones RH1 and streptonigrin were markedly greater and the toxicity of menadione, which is detoxified by NQO1, was ameliorated in the NQ16 line. High levels of NQO1 expression were observed throughout xenograft tumors established from the NQ16 cell line. RH1 treatment was effective at statistically reducing tumor volume in NQ16 xenografts at all of the doses tested (0.1, 0.2, 0.4 mg/kg every day for 5 days), whereas only the highest dose of RH1 resulted in a significant reduction in tumor volume in MDA468 xenografts. CONCLUSIONS: The MDA468/NQ16 isogenic cell line pair is a useful model system for evaluating the role of NQO1 in the bioactivation of antitumor quinones in both cell lines and xenografts. In addition, our data demonstrate that the novel antitumor quinone RH1, is effectively activated by NQO1 both in vitro and in vivo. PMID- 15131057 TI - Decreased nucleotide excision repair activity and alterations of topoisomerase IIalpha are associated with the in vivo resistance of a P388 leukemia subline to F11782, a novel catalytic inhibitor of topoisomerases I and II. AB - PURPOSE: The purpose of the study was to investigate the mechanisms associated with antitumor activity and resistance to F11782, a novel dual catalytic inhibitor of topoisomerases with DNA repair-inhibitory properties. EXPERIMENTAL DESIGN: For that purpose, an F11782-resistant P388 leukemia subline (P388/F11782) has been developed in vivo and characterized. RESULTS: Weekly subtherapeutic doses of F11782 (10 mg/kg) induced complete resistance to F11782 after 8 weekly passages. This resistant P388/F11782 subline retained some in vivo sensitivity to several DNA-topoisomerase II and/or I complex-stabilizing poisons and showed marked collateral sensitivity to cisplatin, topotecan, colchicine, and Vinca alkaloids, while proving completely cross-resistant only to merbarone and doxorubicin. Therefore, resistance to F11782 did not appear to be associated with a classic multidrug resistance profile, as further reflected by unaltered drug uptake and no overexpression of resistance-related proteins or modification of the glutathione-mediated detoxification process. In vivo resistance to F11782 was, however, associated with a marked reduction in topoisomerase IIalpha protein (87%) and mRNA (50%) levels, as well as a diminution of the catalytic activity of topoisomerase IIalpha. In contrast, only minor reductions in topoisomerases IIbeta and I levels were recorded. However, of major interest, nucleotide excision repair activity was decreased 3-fold in these P388/F11782 cells and was more specifically associated with a decreased (67%) level of XPG (human xeroderma pigmentosum group G complementing protein), an endonuclease involved in this DNA repair system. CONCLUSIONS: These findings suggest that both topoisomerase IIalpha and XPG are major targets of F11782 in vivo and further demonstrate the original mechanism of action of this novel compound. PMID- 15131058 TI - Suppression of constitutive and tumor necrosis factor alpha-induced nuclear factor (NF)-kappaB activation and induction of apoptosis by apigenin in human prostate carcinoma PC-3 cells: correlation with down-regulation of NF-kappaB responsive genes. AB - PURPOSE: Development of androgen independence and resistance to apoptosis in prostate cancer are often correlated with high levels of serum tumor necrosis factor (TNF)-alpha in these patients. The loss of sensitivity to TNF-alpha induced apoptosis in androgen-insensitive prostate carcinoma cells is due in part to constitutive activation of Rel/nuclear factor (NF)-kappaB transcription factors that regulate several cell survival and antiapoptotic genes. Our previous studies have demonstrated growth inhibitory and apoptotic effects of apigenin, a common plant flavonoid, in a variety of human prostate carcinoma cells. Here we examined whether apigenin is effective in inhibiting NF-kappaB expression in androgen-insensitive human prostate carcinoma cells exhibiting high constitutive levels of NF-kappaB. EXPERIMENTAL DESIGN: Using androgen-insensitive human prostate carcinoma PC-3 cells, the effect of apigenin was assessed on NF-kappaB activation by electrophoretic mobility shift assay and reporter gene assay. Expression of NF-kappaB subunits p65 and p50, IkappaBalpha, p-IkappaBalpha, in beads kinase assay and NF-kappaB-regulated genes were determined by Western blot analysis. Apoptosis was determined by annexin V/propidium iodide staining after fluorescence-activated cell-sorting analysis. RESULTS: Treatment of cells with 10 40- micro M doses of apigenin inhibited DNA binding and reduced nuclear levels of the p65 and p50 subunits of NF-kappaB. Apigenin inhibited IkappaBalpha degradation and IkappaBalpha phosphorylation and significantly decreased IKKalpha kinase activity. Apigenin also inhibited TNF-alpha-induced activation of NF kappaB via the IkappaBalpha pathway, thereby sensitizing the cells to TNF-alpha induced apoptosis. The inhibition of NF-kappaB activation correlated with a decreased expression of NF-kappaB-dependent reporter gene and suppressed expression of NF-kappaB-regulated genes [specifically, Bcl2, cyclin D1, cyclooxygenase-2, matrix metalloproteinase 9, nitric oxide synthase-2 (NOS-2), and vascular endothelial growth factor]. CONCLUSIONS: Our results indicate that inhibition of NF-kappaB by apigenin may lead to prostate cancer suppression by transcriptional repression of NF-kappaB-responsive genes as well as selective sensitization of prostate carcinoma cells to TNF-alpha-induced apoptosis. PMID- 15131059 TI - Arsenic trioxide-induced death of neuroblastoma cells involves activation of Bax and does not require p53. AB - PURPOSE: On the basis of clinical studies showing that arsenic trioxide (As(2)O(3)), via an apoptotic mechanism, and with minimal toxicity induces complete remission in patients with refractory acute promyelocytic leukemia and that multidrug-resistant and p53-mutated neuroblastoma cells are sensitive to As(2)O(3) both in vitro and in vivo, we searched for molecular mechanisms involved in the As(2)O(3)-induced neuroblastoma cell death. EXPERIMENTAL DESIGN: We have studied the effect of As(2)O(3) on the expression and cellular localization of proteins involved in drug-induced death in two neuroblastoma cell lines with intact p53 and two with mutated p53, the latter two displaying multidrug resistance. RESULTS: As(2)O(3) provoked Bax expression in all tested neuroblastoma cell lines, including SK-N-BE(2) cells with mutated p53 and LA-N-1 cells, which have a deleted p53. In all cell lines exposed to As(2)O(3), p21 Bax was proteolytically cleaved in a calpain-dependent way into the more proapoptotic p18 Bax, which was detected exclusively in a mitochondria-enriched subcellular fraction. As(2)O(3) also caused an increase of cytoplasmic cytochrome c, translocation of antiapoptosis-inducing factor to the nuclei, and a slight activation of caspase 3. However, inhibition of caspase 3 did not prevent cell death, whereas inhibition of Bax cleavage was associated with a decreased As(2)O(3)-induced cell death. CONCLUSIONS: We show that multidrug-resistant neuroblastoma cells die after exposure to As(2)O(3), independent of functional p53, suggesting activation of a cytotoxic pathway different from that induced by conventional chemotherapeutic agents. We further propose that proteolytic activation of Bax is an important event in As(2)O(3)-induced cell death. PMID- 15131060 TI - Transcriptional blocks limit adenoviral replication in primary ovarian tumor. AB - PURPOSE: Despite the success of conditionally replicating adenoviruses in tumor models, clinical success has been limited when they are used as a single modality agent. Overcoming the disparity in efficacy between in vivo animal models and human use is a key hurdle for better conditionally replicating adenovirus therapy in humans. We endeavored to identify biological blocks to adenoviral infection and replication in tumor cells. EXPERIMENTAL DESIGN: We hypothesized that the differences in adenoviral replication between ovarian cancer cell lines and patient tumor samples are the result of a block in viral RNA transcription. To test this hypothesis, established ovarian cancer cell lines and purified patient ovarian cancer cells were infected with wild-type adenovirus. RNA for early adenoviral genes E1A and E1B as well as the late transcripts for fiber and hexon were measured using real-time PCR. RESULTS: Established ovarian cancer cell lines treated with wild-type virus had a lower E1A:E1B ratio than the patient samples. Additionally, the levels of fiber and hexon relative to E1A were also decreased in the patient samples compared with the established cell lines. These findings were consistent with an early- to late-phase block in the adenovirus replication cycle. CONCLUSIONS: These data suggest that the biology of abortive infection in the patient samples may be linked to a defect in the production of early and late viral transcripts. Identification of factors leading to abortive infection will be crucial to understanding the low viral replication in patient samples. PMID- 15131061 TI - Antisense RNA down-regulation of bcl-2 expression in DU145 prostate cancer cells does not diminish the cytostatic effects of G3139 (Oblimersen). AB - PURPOSE: Inhibition of the function of the bcl-2 protein has been postulated to sensitize cells to cytotoxic chemotherapy, and thus provides an attractive target for investigative therapies. G3139, a phosphorothioate antisense oligonucleotide targeted to the initiation codon region of the bcl-2 mRNA, is currently being evaluated in several Phase II and Phase III clinical trials. However, the mechanism of action of this molecule appears to depend on a combination of antisense plus nonantisense events. Indeed, the very idea that bcl-2 is a critical target is, at least in part, an extrapolation from experiments in which intracellular bcl-2 protein concentrations have been dramatically increased, yielding chemoresistant cells. EXPERIMENTAL DESIGN: In this work, we down regulated the expression of bcl-2 protein by 80-90% by two different antisense RNA strategies (antisense RNA and small interfering RNA) in DU145 prostate cancer cells. RESULTS: Even after down-regulation of bcl-2 protein expression by either one of these strategies, the cellular phenotype induced by subsequent G3139 treatment (inhibition of cellular growth and the generation of reactive oxygen species) was essentially identical to that induced in mock-infected or wild-type DU145 cells in which bcl-2 protein expression had not been down-regulated previously. CONCLUSIONS: These results strongly suggest that bcl-2 expression in DU145 cells is not strongly associated with the prolife phenotype and that the mechanism by which G3139 produces its cytostatic effects in this cell line is bcl 2 independent. PMID- 15131062 TI - Induction of cell cycle arrest and apoptosis by the proteasome inhibitor PS-341 in Hodgkin disease cell lines is independent of inhibitor of nuclear factor kappaB mutations or activation of the CD30, CD40, and RANK receptors. AB - PURPOSE: The malignant Hodgkin and Reed-Sternberg cells of Hodgkin disease (HD) are known to constitutively express high levels of activated nuclear factor kappaB (NF-kappaB), which plays an important role in their survival. The proteasome inhibitor PS-341 has been recently shown to modulate tumor cell proliferation and survival by inhibiting NF-kappaB and modulating critical cellular regulatory proteins, but its activity in cells carrying IkappaBalpha gene mutations has not been reported previously. EXPERIMENTAL DESIGN: The activity of PS-341 in four well-characterized, HD-derived cell lines. Cell proliferation and apoptosis were determined by the 3-(4,5-dimethylthiazol-2-yl)-5 (3-carboxymethoxy-phenyl)-2-(4-sulfonyl)-2H-tetrazolium (MTS) and Annexin-V binding methods, respectively. Cell cycle analysis was determined by flow cytometry. Intracellular protein levels were determined by Western blot. RESULTS: PS-341 demonstrated a strong antiproliferative activity, which was irrespective of the status of mutations in IkappaBalpha and even the presence of CD30, CD40, or RANK receptor activation. This effect was attributable to the induction of apoptosis and cell cycle arrest at the G(2)-M phase. PS-341 not only inhibited nuclear localization of NF-kappaB but also activated the caspase cascade, increased p21 and Bax levels, and decreased Bcl-2 levels. Furthermore, PS-341 enhanced the effect of gemcitabine chemotherapy and potentiated the effect of tumor necrosis factor-related apoptosis-inducing ligand/APO2L and two agonistic antibodies to tumor necrosis factor-related apoptosis-inducing ligand death receptors R1 and R2. CONCLUSIONS: The in vitro activity of PS-341 against HD derived cell lines suggests that PS-341 may have a therapeutic value for the treatment of HD. PMID- 15131063 TI - Resistance to tyrosine kinase inhibition by mutant epidermal growth factor receptor variant III contributes to the neoplastic phenotype of glioblastoma multiforme. AB - PURPOSE: We have reported previously that tumors expressing wild-type epidermal growth factor receptor (EGFR) in a murine model are sensitive to the EGFR tyrosine kinase inhibitor gefitinib, whereas tumors expressing mutant EGFR variant III (EGFRvIII) are resistant. Determination of how this differential inhibition occurs may be important to patient selection and treatment criteria, as well as the design of future therapeutics for glioblastoma multiforme. EXPERIMENTAL DESIGN: We have determined and quantified how treatment with gefitinib at commonly used, noncytotoxic doses affects neoplastic functions ascribed to EGFRvIII, including downstream signaling by Akt, DNA synthesis, and cellular invasion. In doing so, we have tested and compared a series of wild-type and mutant EGFRvIII-expressing fibroblast and glioblastoma cell lines in vitro after treatment with gefitinib. RESULTS: The results of these experiments demonstrate that short-term treatment with gefitinib (approximately 24 h) does not reduce phosphorylation of EGFRvIII, whereas EGFR phosphorylation is inhibited in a dose-dependent manner. However, after daily treatment with gefitinib, phosphorylation declines for EGFRvIII by day 3 and later. Nevertheless, after 7 days of daily treatment, cells that express and are dependent on EGFRvIII for tumorigenic growth are not effectively growth inhibited. This may be due in part to phosphorylation of Akt, which is inhibited in EGFR-expressing cells after treatment with gefitinib, but is unaffected in cells expressing EGFRvIII. Cell cycle analysis shows that nascent DNA synthesis in EGFR-expressing cells is inhibited in a dose-dependent manner by gefitinib, yet is unaffected in EGFRvIII expressing cells with increasing dosage. Furthermore, cells expressing EGFRvIII demonstrate greater invasive capability with increasing gefitinib concentration when compared with cells expressing EGFR after treatment. CONCLUSIONS: We conclude that the neoplastic phenotype of EGFRvIII is relatively resistant to gefitinib and requires higher doses, repeated dosing, and longer exposure to decrease receptor phosphorylation. However, this decrease does not effectively inhibit the biologically relevant processes of DNA synthesis, cellular growth, and invasion in cells expressing EGFRvIII. PMID- 15131064 TI - Infection with oncolytic herpes simplex virus-1 induces apoptosis in neighboring human cancer cells: a potential target to increase anticancer activity. AB - PURPOSE: The antitumor efficacy of a herpes simplex virus (HSV)-1 oncolytic virus depends on the cytotoxic effect of the virus, but also on viral replication and spread within the tumor. Apoptosis is considered a defense mechanism of infected cells that minimizes the spread of viral progeny by limiting cellular production of virus. We sought to determine whether oncolytic HSV-1 infection induces apoptosis in neighboring, uninfected cells and whether manipulation of apoptosis can increase viral replication and cytotoxicity. EXPERIMENTAL DESIGN: NV1066 is an oncolytic HSV-1 mutant that contains the marker gene for enhanced green fluorescent protein. OCUM human gastric cancer cells were infected with NV1066 in vitro and inspected for apoptosis by Hoechst and terminal deoxynucleotidyltransferase-mediated nick end labeling staining and for infection by expression of green fluorescence. RESULTS: A significant increase in apoptosis was seen in cells infected by NV1066. More interestingly, a significant percentage (10%) of uninfected cells also proceeded to apoptosis. After NV1066 infection, cells were also treated with N-acetylcysteine (NAC), an inhibitor of apoptosis. By day 4 after infection, 2.7x more NV1066 was produced in cells exposed to NAC than in those not exposed to NV1066 (P = 0.04). NAC also increased tumor kill when administered with virus. CONCLUSIONS: These data suggest that NV1066 induces apoptosis in uninfected cocultured cells, potentially hindering propagation of viral progeny and concomitant tumor kill. Inhibition of apoptosis may improve the efficacy of oncolytic HSV-1 therapy. PMID- 15131065 TI - Effects of a new cystic fibrosis transmembrane conductance regulator inhibitor on Cl- conductance in human sweat ducts. AB - Effective and specific inhibition of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel in epithelia has long been needed to better understand the role of anion movements in fluid and electrolyte transport. Until now, available inhibitors have required high concentrations, usually in the millimolar or high micromolar range, to effect even an incomplete block of channel conductance. These inhibitors, including 5-nitro-2(3-phenylpropyl amino)benzoate (NPPB), bumetamide, glibenclamide and DIDS, are also relatively non-specific. Recently a new anion channel inhibitor, a thiazolidinone derivative, termed CFTRInh-172 has been synthesized and introduced with apparently improved inhibitory properties as shown by effects on anion conductance expressed in cell lines and on secretion in vivo. Here, we assay the effect of this inhibitor on a purely salt absorbing native epithelial tissue, the freshly isolated microperfused human sweat duct, known for its inherently high expression of CFTR. We found that the inhibitor at a maximum dose limited by its aqueous solubility of 5 microm partially blocked CFTR when applied to either surface of the membrane; however, it may be somewhat more effective from the cytosolic side (approximately 70% inhibition). It may also partially inhibit Na+ conductance. The inhibition was relatively slow, with a half time for maximum effect of about 3 min, and showed very slow reversibility. Results also suggest that CFTR Cl- conductance (GCl) was blocked in both apical and basal membranes. The inhibitor appears to exert some effect on Na+ transport as well. PMID- 15131066 TI - Role of nitric oxide in the reflex diuresis in rabbits during pulmonary lymphatic obstruction. AB - The role of nitric oxide in the reflex diuresis in response to pulmonary lymphatic drainage was examined in anaesthetized, artificially ventilated New Zealand White rabbits. Pulmonary lymphatic drainage was obstructed by raising the pressure in a pouch created from the right external jugular vein. Pulmonary lymphatic obstruction resulted in a significant increase in urine flow from an initial control value of 8.9 +/- 0.5 ml (10 min)(-1) to 12.1 +/- 0.6 ml (10 min)( 1) during lymphatic obstruction (mean +/-s.e.m.; n= 17, P < 0.001). This increase in urine flow was accompanied by a significant increase in the excretion of sodium. Additionally, renal blood flow remained unchanged during the increase in urine flow caused by lymphatic obstruction. Intravenous infusion of L-NAME, a non selective inhibitor of nitric oxide synthase (NOS), abolished the reflex diuresis. Furthermore, intraperitoneal administration of the relatively selective neuronal NOS blocker, 7-nitroindazole also abolished the response. It was observed that infusion of a more soluble neuronal NOS blocker, 7-nitroindazole sodium salt (7-NINA), into the renal medulla also abolished the reflex diuresis. These findings suggest that the increase in urine flow in rabbits caused by pulmonary lymphatic obstruction is dependent upon the integrity of neuronal NOS activity within the renal medulla. PMID- 15131067 TI - The baroreflex is counteracted by autoregulation, thereby preventing circulatory instability. AB - The aims of this study were (a) to apply in the animal with intact baroreflex a two-point method for estimation of overall, effective open-loop gain, G0e, which results from the combined action of baroregulation and total systemic autoregulation on peripheral resistance; (b) to predict specific baroreflex gain by correcting the effective gain for the autoregulation gain; and (c) to discuss why the effective gain is usually as low as 1-2 units. G0e was estimated from two measurements of both cardiac output, Q, and mean systemic arterial pressure, P: one in the reference state (set-point) and the other in a steady-state reached 1 3 min after a small cardiac output perturbation. In anaesthetized cats and dogs a cardiac output perturbation was accomplished by partial occlusion of the inferior vena cava and by cardiac pacing, respectively. Average (+/-s.e.m.) estimates of G0e were 1.4 +/- 0.2 (n= 8) in the cat and 1.5 +/- 0.4 (n= 5) in the dog. The specific baroreflex open-loop gain, G0b, found after correction for total systemic autoregulation, was 3.3 +/- 0.4 in the cat and 2.8 +/- 0.8 in the dog. A model-based analysis showed that, with G0e as low as 1.4, the closed-loop response of P to a stepwise perturbation in Q results in damped oscillations that disappear in about 1 min. The amplitude and duration of these oscillations, which have a frequency of about 0.1 Hz, increase with increasing G0e and cause instability when G0e is about 3. We conclude that autoregulation reduces the effectiveness of baroreflex gain by about 55%, thereby preventing instability of blood pressure response. PMID- 15131068 TI - The effects of breathing 5% CO2 on human cardiovascular responses and tolerance to orthostatic stress. AB - Breathing carbon dioxide (CO2) is known to induce hypercapnic acidosis and to affect chemoreceptor regulation of the cardiovascular system. However, there is limited information in the literature regarding the effects of breathing CO2 upon tolerance to orthostatic stress where cardiovascular regulation is challenged. The purpose of this study was to investigate the effect of breathing 5% CO2 on presyncopal tolerance to lower body negative pressure (LBNP). Nine subjects (five males and four females; average +/-s.d. age 21.9 +/- 0.9 years, height 172.4 +/- 9.7 cm, mass 70.3 +/- 7.1 kg) volunteered to participate in this study. Orthostatic tolerance was determined by exposing subjects to LBNP until the onset of presyncopal signs and symptoms on two occasions each separated by approximately 1 week. On one occasion investigations were carried out while subjects were breathing room air and on the other while subjects were breathing air containing 5% CO2, inducing hypercapnia and stimulating systemic chemoreceptors. During hypercapnic conditions, as compared with normocapnia, there were significant increases (P < 0.05) in minute ventilation, end-tidal CO2 and estimated arterial P(CO2). Furthermore, under hypercapnic conditions there was an increase in orthostatic tolerance, peak heart rate and time to peak heart rate during LBNP. The LBNP-induced increase in calf circumference was significantly attenuated at -50 mmHg of LBNP in addition to a further 22.3% reduction in stroke volume under hypercapnic conditions. In conclusion, these results suggest that the possible protective element of presyncope was delayed during hypercapnia at the expense of further reductions in stroke volume. This delayed presyncopal response may have been associated with increases in cerebral blood flow (CBF) induced by the increased arterial P(CO2). PMID- 15131069 TI - Critical appraisal of the mouse model of myocardial infarction. AB - In order to critically evaluate the utility of a mouse model of myocardial infarction (MI) for therapeutic studies, we investigated survival, haemodynamic measurements and histopathology in mice with an occluding suture placed at one of three distinct sites along the left anterior descending coronary artery. The suture was placed at the atrioventricular juncture (High), or at two sites more distally towards the base (Middle and Low). In the High group, only 33% of animals survived 7 days after MI (P < 0.05 compared to all other groups). Only the Middle group had significantly reduced haemodynamics compared to sham operated animals (maximum left ventricular pressure: 55.9 +/- 3.5 versus 80.8 +/- 5.1 mmHg, maximum change in pressure over time : 2003 +/- 172 versus 4402 +/- 491, P < 0.01). Histological examination showed morphological changes in all MI groups. The Middle group had larger lesions than the Low group (P < 0.05). Lesions in the anterior and lateral walls correlated, albeit weakly, with cardiac function. Power calculations indicated that, despite a certain amount of intragroup variation, the Middle Suture model may be useful for therapeutic studies to assess the effects of treatment on cardiac function and overall lesion size. PMID- 15131070 TI - Effects of chronic heart failure in rats on the recovery of microvascular PO2 after contractions in muscles of opposing fibre type. AB - Chronic heart failure (CHF) impairs muscle O2 delivery (QO2) and, at a given O2 uptake (VO2), lowers microvascular O2 pressures (PmvO2: determined by the QO2-to VO2 ratio), which may impair recovery of high-energy phosphates following exercise. Because CHF preferentially decreases QO2 to slow-twitch muscles, we hypothesized that recovery PmvO2 kinetics would be slowed to a greater extent in soleus (SOL: approximately 84% type I fibres) than in peroneal (PER: approximately 14% type I) muscles of CHF rats. PmvO2 dynamics were determined in SOL and PER muscles of control (CON: n= 6; left ventricular end-diastolic pressure, LVEDP: approximately 3 mmHg), moderate CHF (MOD: n= 7; LVEDP: approximately 11 mmHg) and severe CHF (SEV: n= 4; LVEDP: approximately 25 mmHg) following cessation of electrical stimulation (180 s; 1 Hz). In PER, neither the recovery PmvO2 values nor the mean response time (MRT; a weighted average of the time to 63% of the overall response) were altered by CHF (CON: 66.8 +/- 8.0, MOD: 72.4 +/- 11.8, SEV: 69.1 +/- 9.5 s). In marked contrast, SOL PmvO2, at recovery onset, was reduced significantly in the SEV group ( approximately 6 Torr) and PmvO2 MRT was slowed with increased severity of CHF (CON: 45.1 +/- 5.3, MOD: 63.2 +/- 9.4, SEV: 82.6 +/- 12.3 s; P < 0.05 CON vs. MOD and SEV). These data indicate that CHF slows PmvO2 recovery following contractions and lowers capillary O2 driving pressure in slow-twitch SOL, but not in fast-twitch PER muscle. These results may explain, in part, the slowed recovery kinetics (phosphocreatine and VO2) and pronounced fatigue following muscular work in CHF patients. PMID- 15131071 TI - Baroreceptors and the long-term control of blood pressure. AB - The current consensus is that arterial baroreceptors are vitally important in the short term (seconds to minutes) control of mean arterial pressure (MAP) but are unimportant in determining the long-term level of MAP. The latter statement is based primarily on two observations: first, that baroreceptors rapidly reset to the prevailing level of MAP and second, that total baroreceptor denervation has no lasting effect on the average daily MAP, although the variability of MAP is increased dramatically. However, recent studies in intact experimental animals have produced results that suggest baroreceptor resetting may not be as rapid or complete as previously thought. Furthermore, reconsideration of the responses to baroreceptor denervation suggest that the condition may accurately represent responses to short-term baroreceptor unloading but not long-term unloading. Results obtained using a new model of chronic baroreceptor unloading indicate that the condition results in a sustained increase in MAP. These results strongly suggest that the role of baroreceptors in the long term control of MAP needs to be revisited. PMID- 15131072 TI - Catecholamine-induced apoptosis and necrosis in cardiac and skeletal myocytes of the rat in vivo: the same or separate death pathways? AB - High levels of catecholamines are myotoxic but the relative amounts of apoptosis and necrosis have not been established in vivo in cardiac and skeletal muscles. Immunohistochemistry was used to detect and quantify myocyte-specific necrosis (myosin antibody in vivo) and apoptosis (caspase-3 antibody in vitro) in the heart and soleus muscles of male Wistar rats that had received single subcutaneous injections of isoprenaline over the range 1 microg to 5 mg [kg body weight (BW)](-1). Peak myocyte apoptosis occurred 3-6 h after, and necrosis 18 h after, a single injection of 5 mg (kg BW)(-1) isoprenaline in vivo. In the heart myocyte death was mediated through the beta1-adrenergic receptor whereas myocyte death in the soleus muscle was mediated through the beta2-adrenergic receptor. Cardiomyocyte death was heterogeneously distributed throughout the heart, being greatest in the left ventricle (LV) subendocardium and peaking close to the apex, but with significantly more necrosis than apoptosis. Extensive co-localization of caspase-3 and myosin labelling was found in the myocytes of both the heart and the slow-twitch soleus muscle. This, together with similar spatial distributions and responses to catecholamine doses, suggests that either caspase-3 activation occurs in necrotic as well as apoptotic myocytes or that a large proportion of apoptotic myocytes progress to secondary necrosis in vivo. PMID- 15131073 TI - Divergence of renal vascular endothelial growth factor mRNA expression and protein level in post-ischaemic rat kidneys. AB - Vascular endothelial growth factor (VEGF) is a potent regulator of angiogenesis and vascular protection. Synthesis of VEGF is induced by hypoxia and different cytokines including interleukin-6 (IL-6) and interleukin-1beta (IL-1beta). However, post-ischaemic alterations of this growth factor in the kidney are incompletely known. To determine VEGF synthesis in renal ischaemia/reperfusion (I/R) injury unilateral warm ischaemia was induced by cross-clamping the left renal pedicle for 55 min followed by 2 and 24 h of reperfusion (T2 and T24 kidneys; n= 6 in each group). Sham-operated, non-clamped animals served as controls (n= 6). Renal VEGF, IL-6 and IL-1beta mRNA expression were determined by reverse transcription-polymerase chain reaction (RT-PCR). VEGF protein level and distribution were determined by Western blot and immunohistochemical analysis. Immunohistochemistry revealed prominent VEGF staining in the outer medulla of control, T2 and T24 kidneys. VEGF immunoreactivity accumulated at the basolateral area of tubular epithelial cells in T2 kidneys, while it was diffuse in control and T24 kidneys. VEGF protein levels were increased 2- to 3-fold in T2 and T24 kidneys (both P < 0.01 versus controls), while VEGF mRNA expression remained unchanged. IL-6 mRNA expression was increased (P < 0.01 versus controls) in T2 kidneys, while IL-1beta mRNA expression remained unchanged. Increased VEGF protein levels but not mRNA expression suggests that during renal I/R injury VEGF synthesis in kidneys--unlike in other organs--is primarily regulated at a post transcriptional level. As IL-6 mRNA expression increased simultaneously with VEGF protein levels, the post-ischaemic regulation of IL-6 and VEGF synthesis might be interrelated in rat kidney. PMID- 15131074 TI - Vagal reflex actions of atrial natriuretic peptide survive physiological but not pathological cardiac hypertrophy in rat. AB - Atrial natriuretic peptide (ANP) enhances cardiac vagal baroreflexes in normotensive animals. In spontaneously hypertensive rats (SHRs) this effect of ANP was absent. The reflex actions of ANP were preserved if hypertrophy was completely prevented in SHRs. However even a small amount of cardiac hypertrophy, with no hypertension, in SHRs was accompanied by a loss of the reflex bradycardic actions of ANP. In the present study, we investigated whether pathophysiological cardiac hypertrophy, induced by one-kidney, one-clip renovascular hypertension (1K-1C; n = 6), or physiological cardiac hypertrophy induced by chronic spontaneous, wheel-running exercise training (n = 7), similarly prevented vagal reflex actions of ANP. Cardiac baroreceptor-activated bradycardia was measured during rapid ramp increases ( approximately 5 s) in blood pressure after bolus doses of methoxamine or vehicle in conscious, chronically instrumented rats during infusions of ANP (50 pmol kg(-1) min(-1)). Compared with uninephrectomised control rats (n = 10), rats with 1K-1C had cardiac hypertrophy (approximately 55% increase in left ventricle:body weight (LV:BW) ratio; P < 0.05) and blunted vagal baroreflex gain (-0.93 +/- 0.18 versus-0.50 +/- 0.13 beats min(-1) mmHg(-1); P < 0.05). ANP did not augment baroreflex function in 1K-1C. Compared with their sedentary controls (n = 7), exercise-trained rats with cardiac hypertrophy ( approximately 20% increase LV:BW ratio; P < 0.05) also had blunted ramp baroreflex bradycardia (-1.28 +/- 0.23 versus-0.57 +/- 0.09 beats min(-1) mmHg( 1); P < 0.05). In contrast, ANP more than doubled baroreflex bradycardia in exercise-trained rats (P < 0.05). The aetiology of cardiac hypertrophy therefore influenced whether ANP retained its vagal baroreflex enhancing properties. PMID- 15131075 TI - Effect of metabolic cage housing on immunoglobulin A and corticosterone excretion in faeces and urine of young male rats. AB - Six 8-week-old Sprague-Dawley rats were studied for 9 days divided into three periods of 3 days each: before transferral to metabolism cages, during metabolic cage housing and after return to their home cages. Faeces were collected daily when the animals were housed in their home cages and every 6 h when the animals were housed in metabolic cages during which time urine was also collected every 6 h. The rate of weight gain was slightly reduced during the 3 days in metabolic cages and the animals produced significantly larger amounts of faeces when housed in metabolic cages than when housed in their home cages. The total faecal excretion of corticosterone (nanograms excreted per hour per kilogram body weight) and immunoglobulin A (IgA) (milligrams excreted per hour per kg body weight) quantified by enzyme-linked immunosorbent assays (ELISAs) exhibited a clear diurnal rhythm in the metabolic cage. Urinary excretions of corticosterone and IgA also followed a clear diurnal cycle. The mean daily amounts of corticosterone excreted were not significantly affected by cage change and by housing in metabolic cages. However, the excretion of faecal IgA was significantly reduced during the 3 days after the period in metabolic cages. Taken together the results indicate that metabolic cage housing is mildly stressful for young adult male rats. PMID- 15131076 TI - The influence of acute hypothermia on renal function of anaesthetized euthermic and acclimatized rats. AB - Acute hypothermia has a major impact on cardiovascular control and renal function, but the extent to which these can be correlated with and influenced by changes in the altered pattern of sympathetic outflow to the kidneys is unclear. Moreover, it is unknown whether these responses to acute hypothermia are altered by chronic cold exposure and this study aimed to examine these factors. Renal function and renal sympathetic nerve activity (RSNA) were measured in male Wistar rats, euthermic (control) or acclimatized (exposed to progressively lower environmental temperature and photoperiod over 8 weeks), anaesthetized with chloralose/urethane. Reduction of core temperature (Tc) to 25 degrees C caused approximately 40% reduction in heart rate (HR), approximately 10% fall in mean arterial blood pressure (MABP), and decreased glomerular filtration rate (GFR) by approximately 50% and approximately 5% in euthermic and acclimatized rats, respectively. At 25 degrees C, urine flow increased some two-fold and absolute and fractional sodium excretions by 4- to 6-fold in the euthermic rats and to a lesser extent in the cold acclimatized rats, while basal levels of fluid excretion were higher in the acclimatized rats. A loss of pulsatility in the RSNA signal with cooling was seen in both groups. One of the factors contributing to modest hypotension during acute hypothermia is a reduction in RSNA. There was a progressive fall in the proportion of RSNA power at HR frequency with cooling of 20% in euthermic and 80% in acclimatized rats. All variables were restored to basal levels on rewarming in both groups of rats. We conclude that natriuresis and diuresis in euthermic rats during hypothermia is a consequence of a reduction in nephron reabsorption, reduced urine osmolality and possibly altered patterning of RSNA. In acclimatized rats, the response was modified by altered renal haemodynamics and/or hormonal influences induced by chronic cold exposure to minimize the hypothermic stress on renal function. PMID- 15131077 TI - The outcome of ICSI of immature MI oocytes and rescued in vitro matured MII oocytes. AB - BACKGROUND: The use of immature oocytes is limited to cases where these are the only available oocytes, and they are usually only microinjected with sperm after having undergone maturation in vitro. This study compares the outcome of injection of sperm into metaphase I oocytes immediately after their denudation (MI) performed 2 h after their retrieval, with the outcome of injection of sperm into rescued in vitro matured metaphase II (IVM MII) oocytes after their short incubation in routine laboratory conditions. METHODS: ICSI was performed on MI oocytes, rescued IVM MII oocytes and on MI oocytes that were incubated but failed to extrude their first polar body (arrested IVM MI). Fertilization and cleavage rates were compared with those achieved in mature metaphase II oocytes (MII). RESULTS: ICSI of MI oocytes showed impaired performance compared with ICSI of rescued IVM MII oocytes and MII oocytes, in terms of oocyte degeneration rate (11 versus 6 versus 4%; P < 0.0001), fertilization rate (28 versus 44 versus 68%; P < 0.0001) and multipronucleated fertilization (10 versus 4 versus 4%; P < 0.01). The cleavage rate was lower in rescued IVM MII oocytes compared with MII oocytes (86 versus 95%; P < 0.01). Arrested IVM MI oocytes showed similar results to those of MI oocytes but had a lower cleavage rate (72 versus 96%; P < 0.01). CONCLUSIONS: The injection of rescued IVM MII oocytes is preferred to the injection of MI oocytes. PMID- 15131078 TI - Parents' knowledge about the donors and their attitudes toward disclosure in oocyte donation. AB - BACKGROUND: Oocyte donation is a popular treatment option among women with ovarian dysfunction. Little is known about the amount of information recipients have about their donors and if the amount of information the couple has relates to their plans to disclose. The purpose of this study was to assess the amount of information recipients had about their donors and their disclosure plans. METHODS: Sixty-two sets of oocyte donation parents from five programmes completed a self-report questionnaire. RESULTS: Ninety percent of both men and women knew their donor's age, ethnicity, hair colour, eye colour, height, weight, education and medical history. Significantly more women than men told others about using a donor to conceive, but two-thirds of women and men would not tell others if they had to do it over again. Fifty-nine percent of women and 52% of men planned to or had told their child; 34% of women and 41% of men do not plan to tell. The amount of information known about the donor was related to plans to tell the child for men only. CONCLUSION: Approximately half of couples plan to tell their child of their oocyte donor origin and a majority have told others but many regret having done so. Knowledge about the donor is related to disclosure for men only. PMID- 15131079 TI - In utero exposure to organochlorines and age at menarche. AB - BACKGROUND: To examine the effect of in utero exposure to polychlorinated biphenyls (PCBs) and dichlorodiphenyldichloroethylene (DDE) on age at menarche in offspring, we conducted a cohort study over two generations. METHODS: Female participants (and their offspring) in a Michigan angler cohort in which organochlorine levels had been determined previously were studied. Of their 213 female offspring aged 20-50 years, 151 participated in the study (71%). We retrospectively determined age at first menstrual bleeding. Based on repeated maternal serum measurements between 1973 and 1991, we extrapolated PCB and DDE serum levels at the time of pregnancy. To estimate the association between in utero PCB and DDE exposure and age at menarche, we used linear regression analyses controlling for birth date period, maternal age at delivery, birth weight, breastfeeding, education status and maternal height. RESULTS: An increase in the in utero DDE exposure of 15 micro g/l reduced age at menarche by 1 year (P = 0.04). There was no association with maternal PCB exposure. When controlling for estimated body size at menarche, the DDE association was no longer significant, based on a subsample of 102 women. CONCLUSION: The DDE effect on age at menarche encourages further research about in utero exposures. Prospective studies including the offspring's DDE level before menarche are of particular interest. PMID- 15131080 TI - ART in recurrent miscarriage: preimplantation genetic diagnosis/screening or surrogacy? AB - Recently, assisted reproductive techniques have been used to prevent further miscarriages in women with recurrent miscarriage. One approach uses either screening or diagnosis of embryonic chromosomes prior to embryo replacement [preimplantation genetic screening (PGS)/preimplantation genetic diagnosis (PGD)]. The second approach involves surrogacy. However, PGS/PGD assumes that the embryo is chromosomally abnormal, and that the mother should receive a chromosomally normal embryo. Surrogacy assumes that the embryo is normal and that the maternal environment needs to be substituted. This article examines the place of both techniques in different types of recurrent miscarriage, and tries to give guidelines as to which technique is preferable depending on the likelihood of an embryonic chromosome aberration. In repeated fetal aneuploidy or in the older patient, PGS or PGD are preferable. However, with high numbers of miscarriages, or in autoimmune pregnancy loss, surrogacy is preferable. In the light of recent work, it is uncertain which treatment mode is indicated in balanced parental chromosome aberrations. In conclusion, both techniques have a place, but probably only in those patients with a poor prognosis in whom assisted reproductive techniques will be shown to improve the subsequent live birth rate above the spontaneous rate. PMID- 15131081 TI - Holoenzyme proteins required for the physiological assembly and activity of telomerase. AB - Many proteins have been implicated in the physiological function of telomerase, but specific roles of telomerase-associated proteins other than telomerase reverse transcriptase (TERT) remain ambiguous. To gain a more comprehensive understanding of catalytically active enzyme composition, we performed affinity purification of epitope-tagged, endogenously assembled Tetrahymena telomerase. We identified and cloned genes encoding four telomerase proteins in addition to TERT. We demonstrate that both of the two new proteins characterized in detail, p65 and p45, have essential roles in the maintenance of telomere length as part of a ciliate telomerase holoenzyme. The p65 subunit contains an La motif characteristic of a family of direct RNA-binding proteins. We find that p65 in cell extract is associated specifically with telomerase RNA, and that genetic depletion of p65 reduces telomerase RNA accumulation in vivo. These findings demonstrate that telomerase holoenzyme proteins other than TERT play critical roles in RNP biogenesis and function. PMID- 15131082 TI - The action of ARGONAUTE1 in the miRNA pathway and its regulation by the miRNA pathway are crucial for plant development. AB - MicroRNAs (miRNAs) are endogenous 21-24-nt RNAs that can down-regulate gene expression by pairing to the messages of protein-coding genes to specify mRNA cleavage or repression of productive translation. They act within the RNA-induced silencing complex (RISC), which in animals contains a member of the Argonaute family of proteins. In the present study, we show that Arabidopsis ago1 mutants have increased accumulation of mRNAs known to be targeted for cleavage by miRNAs. In hypomorphic ago1 alleles, this compromised miRNA function occurs without a substantial change in miRNA accumulation, whereas in null alleles it is accompanied by a drop in some of the miRNAs. Therefore, AGO1 acts within the Arabidopsis miRNA pathway, probably within the miRNA-programmed RISC, such that the absence of AGO1 destabilizes some of the miRNAs. We also show that targeting of AGO1 mRNA by miR168 is needed for proper plant development, illustrating the importance of feedback control by this miRNA. Transgenic plants expressing a mutant AGO1 mRNA with decreased complementarity to miR168 overaccumulate AGO1 mRNA and exhibit developmental defects partially overlapping with those of dcl1, hen1, and hyl1 mutants showing a decrease in miRNA accumulation. miRNA targets overaccumulate in miR168-resistant plants, suggesting that a large excess of AGO1 protein interferes with the function of RISC or sequesters miRNAs or other RISC components. Developmental defects induced by a miR168-resistant AGO1 mRNA can be rescued by a compensatory miRNA that is complementary to the mutant AGO1 mRNA, proving the regulatory relationship between miR168 and its target and opening the way for engineering artificial miRNAs in plants. PMID- 15131083 TI - Viral RNA silencing suppressors inhibit the microRNA pathway at an intermediate step. AB - RNA silencing suppressors from different plant viruses are structurally diverse. In addition to inhibiting the antiviral silencing response to condition susceptibility, many suppressors are pathogenicity factors that cause disease or developmental abnormalities. Here, unrelated suppressors from multiple viruses were shown to inhibit microRNA (miRNA) activities and trigger an overlapping series of severe developmental defects in transgenic Arabidopsis thaliana. This suggests that interference with miRNA-directed processes may be a general feature contributing to pathogenicity of many viruses. A normally labile intermediate in the miRNA biogenesis/RNA-induced silencing complex (RISC) assembly pathway, miRNA*, accumulated specifically in the presence of suppressors (P1/HC-Pro, p21, or p19) that inhibited miRNA-guided cleavage of target mRNAs. Both p21 and p19, but not P1/HC-Pro, interacted with miRNA/miRNA* complexes and hairpin RNA-derived short interfering RNAs (siRNAs) in vivo. In addition, p21 bound to synthetic miRNA/miRNA* and siRNA duplexes in vitro. We propose that several different suppressors act by distinct mechanisms to inhibit the incorporation of small RNAs into active RISCs. PMID- 15131084 TI - Collaboration of Brca1 and Chk2 in tumorigenesis. AB - Disruption of Brca1 results in cellular demise or tumorigenesis depending on cellular context. Inactivation of p53 contributes to Brca1-associated tumor susceptibility. However the activation of p53-dependent checkpoint/apoptotic signaling in the absence of Brca1 is poorly understood. Here, we show that Chk2 inactivation is partially equivalent to p53 inactivation, in that Chk2 deficiency facilitates the development, survival, and proliferation of Brca1-deficient T cells at the expense of genomic integrity. Brca1 deficiency was found to result in Chk2 phosphorylation and the Chk2-dependent accumulation and activation of p53. Furthermore, inactivation of Chk2 and Brca1 was cooperative in breast cancer. Our findings identify a critical role for Chk2 as a component of the DNA damage-signaling pathway activated in response to Brca1 deficiency. PMID- 15131085 TI - A combined computational-experimental approach predicts human microRNA targets. AB - A new paradigm of gene expression regulation has emerged recently with the discovery of microRNAs (miRNAs). Most, if not all, miRNAs are thought to control gene expression, mostly by base pairing with miRNA-recognition elements (MREs) found in their messenger RNA (mRNA) targets. Although a large number of human miRNAs have been reported, many of their mRNA targets remain unknown. Here we used a combined bioinformatics and experimental approach to identify important rules governing miRNA-MRE recognition that allow prediction of human miRNA targets. We describe a computational program, "DIANA-microT", that identifies mRNA targets for animal miRNAs and predicts mRNA targets, bearing single MREs, for human and mouse miRNAs. PMID- 15131086 TI - Reducing attrition in panel studies in developing countries. AB - BACKGROUND: Panel studies offer repeated observations of individuals over time, but the mobility of populations in the developing world often causes attrition in panel studies. Such attrition can cause bias if it is selective but can be reduced by tracking respondents. Tracking in developing countries can be costly and difficult as populations are often highly mobile, infrastructure is poor, structures frequently change, and formal address systems or population records rarely exist. Method In this paper, the attrition and tracking experiences of panel studies in developing countries are reviewed and recommendations made for ensuring effective tracking. Comments Tracking can reduce attrition by up to 45% and is feasible if procedures are locally appropriate, well planned, involve the community, collect as much locating data as possible, and have explicit criteria, and if tracking is done at regular intervals, and interviewers are well trained, supervised, and motivated. CONCLUSION: Attrition is an important issue in panel studies, whilst tracking can be costly it can reduce attrition if effective procedures are used. PMID- 15131087 TI - Anxiety or depressive disorders and risk of ischaemic heart disease among French power company employees. AB - BACKGROUND: The aim of this study was to determine whether depression and anxiety are predictive factors for ischaemic heart disease among adults <60 years. Method A case-control study among active employees of the French nationwide power company (Electricite de France-Gaz de France) analysed men aged 31-55 years who presented an initial clinical form of ischaemic heart disease from 1993 through 1997, collected from the company registry. These 660 men were each matched by age to 10 controls per case. Adjusted odds ratios (OR) were calculated by logistic regression. RESULTS: There was a significant association between ischaemic heart disease and sick-leave for any medical reason in the 3 years before its onset (OR = 1.79; 95% CI: 1.50, 2.14). This association was strengthened when only absences for depression and anxiety were considered (OR = 3.10; 95% CI: 2.29, 4.19) and remained important and significant when adjusted for socioeconomic status: OR = 2.66 (95% CI: 1.95, 3.63). A previous sick-leave for depression or anxiety in the 10 years before the heart disease strengthened the association (OR = 3.61; 95% CI: 2.39, 4.45), which was further reinforced by an elevated number (> or =4) of such sick-leaves (OR = 5.11; 95 % CI: 3.11, 8.40). CONCLUSION: Depressive and anxiety disorders that lead to absenteeism seem to be associated with an increased risk of ischaemic heart disease in the 3 years thereafter, especially when depression and anxiety were severe and chronic; this association is independent of socioeconomic status. PMID- 15131088 TI - Are patients with asthma at increased risk of coronary heart disease? AB - BACKGROUND: Inflammation plays a role in the pathogenesis of athero-thrombosis. Because of the chronic, inflammatory nature of asthma, we hypothesized a possible link asthma and prospective risk of coronary heart disease (CHD). METHODS: We performed a cohort study among 70 047 men and 81 573 women, 18-85 years old, enrolled in a large managed care organization in Northern California. Asthma was ascertained by self-report at baseline in 1964-1973 and/or interim hospitalization for asthma during follow-up. The primary endpoint was combined non-fatal or fatal CHD. RESULTS: After a median follow-up time of 27 years, and adjusting for age, race/ethnicity, education level, smoking status, alcohol consumption, body mass index, serum total cholesterol, white blood cell count, hypertension, diabetes, and history of occupational exposures, asthma was associated with a 1.22-fold (95% CI: 1.14, 1.31) increased hazard of CHD among women. This association was seen both in never and in ever smoking women, and in younger and older women. By contrast, asthma was not associated with CHD among men (multivariate-adjusted hazard ratio = 0.99; 95% CI: 0.93, 1.05). CONCLUSIONS: Asthma was independently associated with a modest but statistically significant increased hazard of CHD among women. Further studies are warranted to confirm or refute these preliminary epidemiological findings. PMID- 15131089 TI - Mortality and educational level among diabetic and non-diabetic population in the Turin Longitudinal Study: a 9-year follow-up. AB - BACKGROUND: People in a lower social position have a higher prevalence of unhealthy behaviour, more difficult access to healthcare, and lower compliance with drug treatment; as a consequence, social differences in mortality are likely to be higher in people with diabetes compared with the non-diabetic population. We compared diabetics with non-diabetics in terms of mortality and social differences in mortality. METHODS: In all, 31 264 residents in Turin (northern Italy), who were > or =20 years old, registered in the local diabetes register between 1991 and 1999. They were followed up from recruitment to December 1999, and their cause-specific mortality by educational level was analysed. This was compared with that of the local non-diabetic population. Diabetes was classified as type 1 (< or =35 years at diagnosis) or type 2 (>35 years). RESULTS: For type 1 diabetes, the all-cause standardized mortality ratio (SMR) was 197.6 (95% CI:155.7, 247.4) in men and 336.0 (95% CI:259.3, 428.2) in women; for type 2 diabetes, the all-cause SMR was 142.8 (95% CI:138, 147.6) in men and 143.4 (95% CI:138.5, 148.5) in women. Whereas social differences in mortality were evident among non-diabetic men and women for all causes of death considered, no significant differences were found among diabetic women. Mortality was slightly increased among less educated diabetic men, particularly for neoplasms, although this gradient was less steep than that among non-diabetics. CONCLUSIONS: These results suggest that the regular clinical follow-up and health education provided by the local network of diabetic centres might play an important role in confronting the adverse effects of diabetes and in reducing social differences in health. PMID- 15131091 TI - Epidemiology in the era of globalization: skills transfer or new skills? AB - BACKGROUND: Globalization carries information and technology opportunities and risks in widened inequalities, a resurgence of old health risks and reversal of health gains. METHODS: The paper explores the implications for epidemiological work in southern Africa and through two case studies-occupational health and equity in health-profiles challenges faced in that region. RESULTS: Occupational epidemiology is confronted by weak monitoring and regulatory systems, healthy worker effects, surveillance filters, and migration. Occupational disease determinants are masked by the combined effects of work, wider environmental risks, and high poverty-related disease. Health burdens associated with new production and trade patterns are thus largely unrecognized. Even when made visible, they may be ignored by economically vulnerable states and workers. Work on equity in health indicates the relevance of social and political determinants in the distribution of health resources. The shift of the cost burden of human immunodeficiency virus (HIV)/AIDS to poor communities and the weak public health response to HIV/AIDS suggest that economic and health reforms associated with globalization have both increased health inequalities and weakened social and political forces promoting equity and solidarity values in public health. CONCLUSIONS: Epidemiology can demystify disease sources and explain determinants in a manner that impacts on public policy and action. Under current conditions of globalization this implies addressing methodological challenges and enhancing uptake of evidence in policy processes. Given the intensifying political struggle around health resources, increased attention needs to be given to participatory forms of inquiry that strengthen the influence of poor communities and public interest values in health policy. PMID- 15131092 TI - Commentary: Does the presence of asthma increase the incidence of coronary heart disease? PMID- 15131093 TI - Commentary: still dying of ignorance? Human immunodeficiency virus (HIV) prevention strategies revisited. PMID- 15131094 TI - Commentary: Socioeconomic status and diabetes outcomes; what might we expect and why don't we find it? PMID- 15131095 TI - Ovariectomy vs. ovariohysterectomy. PMID- 15131096 TI - Use of continuous blood glucose monitoring for animals with diabetes mellitus. PMID- 15131097 TI - Chronic canine ehrlichiosis (Ehrlichia canis): a retrospective study of 19 natural cases. AB - Nineteen dogs from Greece with chronic ehrlichiosis were studied. The dogs exhibited bicytopenia or pancytopenia, bone marrow hypoplasia, seroreactivity to Ehrlichia canis (E. canis) antigens, and had no history of drug or radiation exposure. Anorexia, depression, severe bleeding tendencies, hypoalbuminemia, and increased serum alanine aminotransferase activity were also hallmarks of the disease. All these animals eventually died, irrespective of the treatment applied. Some dogs were also serologically positive for Rickettsia conorii, Leishmania infantum (L. infantum), and Bartonella vinsonii subspp. berkhoffii. Polymerase chain reaction testing of bone marrow samples revealed E. canis, Anaplasma phagocytophilia, Anaplasma platys, and L. infantum in some dogs. Concurrent infections did not appear to substantially influence the clinical course and final outcome of the chronic canine ehrlichiosis. PMID- 15131098 TI - Effects of diet on urine composition of cats with calcium oxalate urolithiasis. AB - Ten client-owned cats with calcium oxalate (CaOx) urolithiasis were evaluated to determine the effect of diet on urine CaOx saturation. Two dietary treatments were evaluated in each cat: the diet consumed just prior to urolith detection and a canned diet formulated to prevent CaOx uroliths. This study revealed that hypercalciuria is a consistent abnormality in cats with CaOx urolith formation. When urolith-forming cats consumed a diet formulated to prevent urolith formation, activity product ratios for CaOx (which estimate the degree to which urine is saturated with CaOx) were significantly lower. These results suggest that consumption of an appropriately formulated urolith-prevention diet will reduce recurrence of CaOx urolithiasis. PMID- 15131099 TI - Diet-related risk factors for gastric dilatation-volvulus in dogs of high-risk breeds. AB - A nested case-control study was conducted among 1634 dogs with complete diet information in a 5-year prospective study to determine diet-related risk factors for gastric dilatation-volvulus (GDV). Cases included 106 dogs that developed GDV; controls included 212 dogs without GDV that were frequency matched to cases by year of GDV onset. Proportionate energy consumed from major food types and from carbohydrates was determined. Dogs were categorized as consuming either a low volume or high volume of food based on the median number of cups of food fed per kg of body weight per meal. Dogs fed a larger volume of food per meal were at a significantly (P<0.05) increased risk of GDV, regardless of the number of meals fed daily. For both large- and giant-breed dogs, the risk of GDV was highest for dogs fed a larger volume of food once daily. PMID- 15131100 TI - Outcomes and complications associated with ventral screws, pins, and polymethyl methacrylate for atlantoaxial instability in 12 dogs. AB - Clinical outcomes and complications of a technique used for atlantoaxial stabilization were evaluated in a group of 12 dogs. At surgery, the atlantoaxial joint was realigned and rigidly fixated using cortical bone screws, K-wire, and polymethyl methacrylate. Results in nine dogs were graded as excellent. Results in two dogs were judged as good. One dog was euthanized 17 months after surgery for recurrent cervical pain. Eight dogs had no postoperative complications. The surgical technique described provided an adaptable method for the correction of atlantoaxial instability. PMID- 15131101 TI - Transnasal laryngoscopy for the diagnosis of laryngeal paralysis in dogs. AB - Four dogs with clinical signs of laryngeal paralysis and three normal dogs were evaluated with transnasal laryngoscopy. Six of these dogs subsequently underwent standard laryngoscopy. For transnasal laryngoscopy, a video endoscope was passed through the left nasal passage after intramuscular sedation and topical anesthesia. The laryngeal opening was observed during spontaneous ventilation. Laryngeal paralysis was diagnosed in four dogs and was confirmed with traditional laryngoscopy in three dogs. Normal motion of the arytenoid cartilages was present in the other three dogs; however, two required mechanical stimulation of the laryngeal mucosa for full evaluation. Transnasal laryngoscopy provided a means for diagnosing laryngeal paralysis in dogs without general anesthesia. PMID- 15131102 TI - Surgical treatment of recurrent dacryocystitis secondary to cystic dilatation of the nasolacrimal duct in a dog. AB - A 3-year-old, castrated male golden retriever was presented for evaluation of recurrent ocular discharge of 4 months' duration from the left eye. Dacryocystorhinography was performed and demarcated a cystic dilatation of the left nasolacrimal duct with obstruction of the duct distal to the cystic cavity. Surgical exploration of the left maxillary sinus was performed to confirm the diagnosis and reestablish drainage into the nasal cavity. Recovery was uneventful, and the dog has been asymptomatic for >36 months postoperatively. This report documents the third published case of surgical treatment for cystic dilatation of the nasolacrimal duct. PMID- 15131103 TI - Spontaneous midlobar lung lobe torsion in a 2-year-old Newfoundland. AB - A 2-year-old, intact male Newfoundland was presented for evaluation of a 1- to 2 day history of coughing, retching, and progressive lethargy. Thoracic radiographs demonstrated right-sided pleural effusion and lobar vesicular emphysema. An exploratory thoracotomy revealed midlobar torsion of the right caudal lung lobe. A partial lung lobectomy was performed, and following surgery, the dog recovered without complications. Lung lobe torsions in dogs typically occur at the hilus of the affected lung lobe. This report documents that midlobar lung lobe torsions occur in dogs and should be considered as a differential diagnosis in cases of lobar vesicular emphysema and pleural effusion. PMID- 15131104 TI - Bone marrow hypoplasia associated with fenbendazole administration in a dog. AB - A 1.5-year-old Doberman pinscher was presented with sudden-onset of fever and malaise. Twelve days prior to presentation, fenbendazole therapy was initiated for a suspected lungworm infection. Results of a complete blood count on presentation showed pancytopenia, while histopathological evaluation of a bone marrow core sample revealed bone marrow hypoplasia of undetermined etiology. Bactericidal antibiotics and fluid therapy, as well as discontinuation of fenbendazole administration, led to a complete resolution of clinical and hematological abnormalities within 15 days. An idiosyncratic reaction to fenbendazole was suspected based on the absence of infectious, neoplastic, autoimmune, and toxic etiologies, as well as resolution of clinical signs and pancytopenia upon drug withdrawal. PMID- 15131105 TI - Phenobarbital-responsive ptyalism, dysphagia, and apparent esophageal spasm in a German shepherd puppy. AB - A 10-week-old, male German shepherd dog was presented with a primary complaint of episodic ptyalism, dysphagia, vomiting, and mandibular salivary gland enlargement. An esophagram with fluoroscopy showed normal pharyngeal and esophageal function; however, upper gastrointestinal endoscopy and cervical ultrasonography revealed a focal circumferential thickening of the midcervical esophageal muscular wall, consistent with esophageal spasm. The puppy responded dramatically and completely to phenobarbital treatment. An unusual syndrome of phenobarbital-responsive hypersialosis was consistent with this dog's clinical presentation and the finding of apparent esophageal spasm. The pathogenesis of this syndrome is unclear, but it may represent a form of limbic epilepsy or peripheral autonomic dysfunction. PMID- 15131106 TI - Exocrine pancreatic neoplasia in the cat: a case series. AB - Eight cases of feline pancreatic adenocarcinoma and two cases of pancreatic adenoma were reviewed. The adenomas were incidental findings. Most cats with adenocarcinomas had anorexia (75%) and vomiting (63%), while 38% had abdominal pain, a palpable abdominal mass, and/or jaundice. Diagnostic abnormalities included leukocytosis, hyperglycemia, increased alanine aminotransferase activity, poor serosal detail on abdominal radiography, and an abdominal mass effect on ultrasonography. The majority of cats with carcinomas had metastases (mostly to liver, lung, and small intestine), and all were euthanized or died within 7 days of diagnosis. Clinically, feline pancreatic carcinoma may be difficult to distinguish from feline pancreatitis. PMID- 15131107 TI - Osteosarcoma of the humeral head associated with osteochondritis dissecans in a dog. AB - A 6-year-old, male Labrador retriever was presented for lameness of the right forelimb. The lameness occurred 5 years after the dog was surgically treated for osteochondritis dissecans (OCD) of the caudal right humeral head. Radiographs revealed an expansile, osteolytic lesion occupying the entire caudal half of the right humeral head, and biopsy confirmed the diagnosis of osteosarcoma. Forelimb amputation was performed. Histopathology of the lesion and associated articular cartilage revealed neoplastic osteoblasts extensively infiltrating the marrow space adjacent to scar tissue associated with the previous OCD lesion. This is the first report of an epiphyseal osteosarcoma in the area of a previous OCD lesion in a dog. PMID- 15131108 TI - Methionine 129 variant of human prion protein oligomerizes more rapidly than the valine 129 variant: implications for disease susceptibility to Creutzfeldt-Jakob disease. AB - The human PrP gene (PRNP) has two common alleles that encode either methionine or valine at codon 129. This polymorphism modulates disease susceptibility and phenotype of human transmissible spongiform encyphalopathies, but the molecular mechanism by which these effects are mediated remains unclear. Here, we compared the misfolding pathway that leads to the formation of beta-sheet-rich oligomeric isoforms of the methionine 129 variant of PrP to that of the valine 129 variant. We provide evidence for differences in the folding behavior between the two variants at the early stages of oligomer formation. We show that Met(129) has a higher propensity to form beta-sheet-rich oligomers, whereas Val(129) has a higher tendency to fold into alpha-helical-rich monomers. An equimolar mixture of both variants displayed an intermidate folding behavior. We show that the oligomers of both variants are initially a mixture of alpha- and beta-rich conformers that evolve with time to an increasingly homogeneous beta-rich form. This maturation process, which involves no further change in proteinase K resistance, occurs more rapidly in the Met(129) form than the Val(129) form. Although the involvement of such beta-rich oligomers in prion pathogenesis is speculative, the misfolding behavior could, in part, explain the higher susceptibility of individuals that are methionine homozygote to both sporadic and variant Creutzfeldt-Jakob disease. PMID- 15131109 TI - Oleic acid modulates the post-translational glycosylation of macrophage ApoE to increase its secretion. AB - There has been increasing interest in a potential role for fatty acids in adversely affecting organismal substrate utilization and contributing to the cardiovascular complications in insulin resistance. Fatty acids have already been implicated in regulating the expression of a number of genes in resident cells of the vessel wall. In the current studies, we evaluated a potential role for fatty acids in the regulation of macrophage apoE expression. Incubation in oleic acid increased the synthesis and secretion of apoE by human monocyte-derived macrophages. Part of this stimulation was mediated at a post-translational locus. Oleic acid increased the secretion of apoE from macrophages that constitutively expressed a human apoE3 cDNA. Incubation in palmitic acid decreased apoE secretion from these cells. The effect of oleic acid on apoE secretion could not be accounted for by the known effect of fatty acid on cellular sterol, because incubation in oleic acid did not suppress the degradation of nascent apoE. Incubation in oleic acid for at least 6 h was required to observe an effect on apoE secretion. Oleic acid altered the glycosylation pattern of cellular and secreted apoE, with a loss of the most heavily sialylated isoform. Oleic acid had no effect on the glycosylation of interleukin 6 secreted from macrophages. Elimination of apoE glycosylation, by substitution of threonine 194 with alanine, eliminated oleic acid-mediated stimulation of apoE secretion. These results indicate that oleic acid increases apoE secretion from macrophages at a locus involving post-translational glycosylation. PMID- 15131111 TI - Structural and kinetic analyses of the interaction of anthrax adenylyl cyclase toxin with reaction products cAMP and pyrophosphate. AB - Anthrax edema factor (EF) raises host intracellular cAMP to pathological levels through a calcium-calmodulin (CaM)-dependent adenylyl cyclase activity. Here we report the structure of EF.CaM in complex with its reaction products, cAMP and PP(i). Mutational analysis confirmed the interaction of EF with cAMP and PP(i) as depicted in the structural model. While both cAMP and PP(i) have access to solvent channels to exit independently, PP(i) is likely released first. EF can synthesize ATP from cAMP and PP(i), and the estimated rate constants of this reaction at two physiologically relevant calcium concentrations were similar to those of adenylyl cyclase activity of EF. Comparison of the conformation of adenosine in the structures of EF.CaM.cAMP.PP(i) with EF.CaM.3.dATP revealed about 160 degrees rotation in the torsion angle of N-glycosyl bond from the +anti conformation in 3.dATP to -syn in cAMP; such a rotation could serve to distinguish against substrates with the N-2 amino group of purine. The catalytic rate of EF for ITP was about 2 orders of magnitude better than that for GTP, supporting the potential role of this rotation in substrate selectivity of EF. The anomalous difference Fourier map revealed that two ytterbium ions (Yb(3+)) could bind the catalytic site of EF.CaM in the presence of cAMP and PP(i), suggesting the presence of two magnesium ions at the catalytic site of EF. We hypothesize that EF could use a "histidine and two-metal ion" hybrid mechanism to facilitate the cyclization reaction. PMID- 15131110 TI - NCB5OR is a novel soluble NAD(P)H reductase localized in the endoplasmic reticulum. AB - The NAD(P)H cytochrome b5 oxidoreductase, Ncb5or (previously named b5+b5R), is widely expressed in human tissues and broadly distributed among the animal kingdom. NCB5OR is the first example of an animal flavohemoprotein containing cytochrome b5 and chrome b5 reductase cytodomains. We initially reported human NCB5OR to be a 487-residue soluble protein that reduces cytochrome c, methemoglobin, ferricyanide, and molecular oxygen in vitro. Bioinformatic analysis of genomic sequences suggested the presence of an upstream start codon. We confirm that endogenous NCB5OR indeed has additional NH2-terminal residues. By performing fractionation of subcellular organelles and confocal microscopy, we show that NCB5OR colocalizes with calreticulin, a marker for endoplasmic reticulum. Recombinant NCB5OR is soluble and has stoichiometric amounts of heme and flavin adenine dinucleotide. Resonance Raman spectroscopy of NCB5OR presents typical signatures of a six-coordinate low-spin heme similar to those found in other cytochrome b5 proteins. Kinetic measurements showed that full-length and truncated NCB5OR reduce cytochrome c actively in vitro. However, both full-length and truncated NCB5OR produce superoxide from oxygen with slow turnover rates: kcat = approximately 0.05 and approximately 1 s(-1), respectively. The redox potential at the heme center of NCB5OR is -108 mV, as determined by potentiometric titrations. Taken together, these data suggest that endogenous NCB5OR is a soluble NAD(P)H reductase preferentially reducing substrate(s) rather than transferring electrons to molecular oxygen and therefore not an NAD(P)H oxidase for superoxide production. The subcellular localization and redox properties of NCB5OR provide important insights into the biology of NCB5OR and the phenotype of the Ncb5or-null mouse. PMID- 15131112 TI - Genome-wide mapping of in vivo targets of the Drosophila transcription factor Kruppel. AB - Kruppel (Kr), a member of the gap class of Drosophila segmentation genes, encodes a DNA binding zinc finger-type transcription factor. In addition to its segmentation function at the blastoderm stage, Kruppel also plays a critical role in organ formation during later stages of embryogenesis. To systematically identify in vivo target genes of Kruppel, we isolated DNA fragments from the Kruppel-associated portion of chromatin and used them to find and map Kruppel dependent cis-acting regulatory sites in the Drosophila genome. We show that Kruppel binding sites are not enriched in Kruppel-associated chromatin and that the clustering of Kruppel binding sites, as found in the cis-acting elements of Kruppel-dependent segmentation genes used for in silico searches of Kruppel target genes, is not a prerequisite for the in vivo binding of Kruppel to its regulatory elements. Results obtained with the newly identified target gene ken and barbie (ken) indicate that Kruppel represses transcription and thereby restricts the spatial expression pattern of ken during blastoderm and gastrulation. PMID- 15131113 TI - Construction of a dual chain pseudotetrameric chicken avidin by combining two circularly permuted avidins. AB - Two distinct circularly permuted forms of chicken avidin were designed with the aim of constructing a fusion avidin containing two biotin-binding sites in one polypeptide. The old N and C termini of wild-type avidin were connected to each other via a glycine/serine-rich linker, and the new termini were introduced into two different loops. This enabled the creation of the desired fusion construct using a short linker peptide between the two different circularly permuted subunits. The circularly permuted avidins (circularly permuted avidin 5 --> 4 and circularly permuted avidin 6 --> 5) and their fusion, pseudotetrameric dual chain avidin, were biologically active, i.e. showed biotin binding, and also displayed structural characteristics similar to those of wild-type avidin. Dual chain avidin facilitates the development of dual affinity avidins by allowing adjustment of the ligand-binding properties in half of the binding sites independent of the other half. In addition, the subunit fusion strategy described in this study can be used, where applicable, to modify oligomeric proteins in general. PMID- 15131114 TI - Minimal structural rearrangement of the cytoplasmic pore during activation of the 5-HT3A receptor. AB - Ligand-gated ion channel receptors mediate the response of fast neurotransmitters by opening in less than a millisecond. Here, we investigated the activation mechanism of a serotonin-gated receptor (5-HT(3A)) by systematically introducing cysteine substitutions throughout the pore-lining M1-M2 loop and M2 transmembrane domain. We hypothesized that multiple cysteines in the narrowest region of the pore, which together can form a high affinity binding site for metal cations, would reveal changes in pore structure during gating. Using cadmium (Cd2+) as a probe, two cysteine substitutions in the cytoplasmic selectivity filter, S2'C and, to a lesser extent, G-2'C, showed high affinity inhibition with Cd2+ when applied extracellularly in the open state. Cd2+ inhibition in S2'C was attenuated if applied in the presence of an open-channel inhibitor and showed voltage dependent recovery, indicating a direct effect of Cd2+ in the pore. When applied intracellularly, Cd2+ appeared to bind S2'C receptors in the closed state. The ability of cysteine side chains at the 2' and -2' positions to coordinate Cd2+ in both the native open and closed states of the channel suggests that the cytoplasmic selectivity filter of 5-HT(3A) receptors maintains a narrow pore during channel gating. PMID- 15131115 TI - Platelet factor XIII and calpain negatively regulate integrin alphaIIbbeta3 adhesive function and thrombus growth. AB - Excessive accumulation of platelets at sites of athero-sclerotic plaque rupture leads to the development of arterial thrombi, precipitating clinical events such as the acute coronary syndromes and ischemic stroke. The major platelet adhesion receptor glycoprotein (GP) IIb-IIIa (integrin alpha(IIb)beta3) plays a central role in this process by promoting platelet aggregation and thrombus formation. We demonstrate here a novel mechanism down-regulating integrin alpha(IIb)beta3 adhesive function, involving platelet factor XIII (FXIII) and calpain, which serves to limit platelet aggregate formation and thrombus growth. This mechanism principally occurs in collagen-adherent platelets and is induced by prolonged elevations in cytosolic calcium, leading to dramatic changes in platelet morphology (membrane contraction, fragmentation, and microvesiculation) and a specific reduction in integrin alpha(IIb)beta3 adhesive function. Adhesion receptor signal transduction plays a major role in the process by sustaining cytosolic calcium flux necessary for calpain and FXIII activation. Analysis of thrombus formation on a type I fibrillar collagen substrate revealed an important role for FXIII and calpain in limiting platelet recruitment into developing aggregates, thereby leading to reduced thrombus formation. These studies define a previously unidentified role for platelet FXIII and calpain in regulating integrin alpha(IIb)beta3 adhesive function. Moreover, they demonstrate the existence of an autoregulatory feedback mechanism that serves to limit excessive platelet accumulation on highly reactive thrombogenic surfaces. PMID- 15131116 TI - DNA methylation polymorphisms precede any histological sign of atherosclerosis in mice lacking apolipoprotein E. AB - The present work investigates the occurrence and significance of aberrant DNA methylation patterns during early stages of atherosclerosis. To this end, we asked whether the genetically atherosclerosis-prone APOE-null mice show any changes in DNA methylation patterns before the appearance of histologically detectable vascular lesion. We exploited a combination of various techniques: DNA fingerprinting, in vitro methyl-accepting assay, 5-methylcytosine quantitation, histone post-translational modification analysis, Southern blotting, and PCR. Our results show that alterations in DNA methylation profiles, including both hyper- and hypomethylation, were present in aortas and PBMC of 4-week-old mutant mice with no detectable atherosclerotic lesion. Sequencing and expression analysis of 60 leukocytic polymorphisms revealed that epigenetic changes involve transcribed genic sequences, as well as repeated interspersed elements. Furthermore, we showed for the first time that atherogenic lipoproteins promote global DNA hypermethylation in a human monocyte cell line. Taken together, our results unequivocally show that alterations in DNA methylation profiles are early markers of atherosclerosis in a mouse model and may play a causative role in atherogenesis. PMID- 15131117 TI - Selection of poly-alpha 2,8-sialic acid mimotopes from a random phage peptide library and analysis of their bioactivity. AB - Poly-alpha 2-8 sialic acid (PSA), attached to the neural cell adhesion molecule, is a permissive determinant for numerous morphogenetic and neural plasticity processes, making it a potential therapeutic target. Here, using a monoclonal antibody specific for PSA, we screened a phage-display library and identified two cyclic nine-amino acid peptides (p1, p2) that are PSA epitope analogues. We evaluated their bioactivity in vitro and in vivo. In culture, micromolar concentrations of the peptides promoted axon growth, defasciculation, and migration of neural progenitors. When injected into developing chicken retina, the peptides modified the trajectory of retinal ganglion cell axons. Moreover, they enhanced migration of grafted neuroblasts in mouse brain. These effects were selective and dependent upon the presence of PSA on transplanted cells. Our results demonstrate the feasibility and therapeutic potential of enhancing PSA biological activity. PMID- 15131118 TI - Characterization of the conformational state and flexibility of HIV-1 glycoprotein gp120 core domain. AB - gp120 is key to the human immunodeficiency virus type 1 viral cell entry. Knowledge of the detailed conformational states of gp120 is crucial to intervention, yet the unbound form is still resistant to structural characterization probably because of its flexibility. Toward this goal, we performed molecular dynamics simulations on the wild type gp120 core domain extracted from its ternary crystal structure and on a modeled mutant, S375W, that experimentally has a significantly different phenotype from the wild type. Although the mutant retained a bound-like conformation, the wild type drifted to a different conformational state. The wild type beta strands 2 and 3 of the bridging sheet were very mobile and partially unfolded, and the organization among the inner and outer domains and beta strands 20 and 21 of the bridging sheet, near the mutation site, was more open than in the bound form, although the overall structure was maintained. These differences were apparently a result of the strengthening of the hydrophobic core in the mutant. This stabilization further explains the experimentally significantly different thermodynamic properties between the wild type and the mutant. Taken together, our results suggest that the free form, although different from the bound state, shares many of the bound structural features. The observed loss of freedom near the binding site, rather than the previously hypothesized more dramatic conformational transition from the unbound to the bound state, appears to be the major contributor to the large entropy cost for the CD4 binding to the wild type. PMID- 15131119 TI - Intrinsic heterogeneity in adipose tissue of fat-specific insulin receptor knock out mice is associated with differences in patterns of gene expression. AB - Mice with a fat-specific insulin receptor knock-out (FIRKO) have reduced adipose tissue mass, are protected against obesity, and have an extended life span. White adipose tissue of FIRKO mice is also characterized by a polarization into two major populations of adipocytes, one small (<50 microm) and one large (>100 microm), which differ with regard to basal triglyceride synthesis and lipolysis, as well as in the expression of fatty acid synthase, sterol regulatory element binding protein 1c, and CCAAT/enhancer-binding protein alpha (C/EBP-alpha). Gene expression analysis using RNA isolated from large and small adipocytes of FIRKO and control (IR lox/lox) mice was performed on oligonucleotide microarrays. Of the 12,488 genes/expressed sequence tags represented, 111 genes were expressed differentially in the four populations of adipocytes at the p < 0.001 level. These alterations exhibited 10 defined patterns and occurred in response to two distinct regulatory effects. 63 genes were identified as changed in expression depending primarily upon adipocyte size, including C/EBP-alpha, C/EBP-delta, superoxide dismutase 3, and the platelet-derived growth factor receptor. 48 genes were regulated primarily by impairment of insulin signaling, including transforming growth factor beta, interferon gamma, insulin-like growth factor I receptor, activating transcription factor 3, aldehyde dehydrogenase 2, and protein kinase Cdelta. These data suggest an intrinsic heterogeneity of adipocytes with differences in gene expression related to adipocyte size and insulin signaling. PMID- 15131120 TI - Role of insulin action and cell size on protein expression patterns in adipocytes. AB - Mice with a fat-specific insulin receptor knock-out (FIRKO) exhibit a polarization of white adipose tissue into two populations of cells, one small (diameter <50 microm) and one large (diameter >100 microm), accompanied by changes in insulin-stimulated glucose uptake, triglyceride synthesis, and lipolysis. To characterize these subclasses of adipocytes, we have used a proteomics approach in which isolated adipocytes from FIRKO and control (IR lox/lox) mice were separated by size, fractionated into cytosolic and membrane subfractions, and analyzed by sucrose gradient, SDS-PAGE, and mass spectrometry. A total of 27 alterations in protein expression at key steps in lipid and energy metabolism could be defined, which were coordinately regulated by adipocyte cell size, impaired insulin signaling, or both. Nine proteins, including vimentin, EH domain-containing protein 2, elongation factor 2, glucose-regulated protein 78, transketolase, and succinyl-CoA transferase were primarily affected by presence or absence of insulin signaling, whereas 21 proteins, including myosin non-muscle form A, annexin 2, annexin A6, and Hsp47 were regulated in relation to adipocyte size. Of these 27 alterations in protein expression, 14 changes correlated with altered levels of mRNA, whereas the remaining 13 were the result of changes in protein translation or turnover. These data suggest an intrinsic heterogeneity in adipocytes with differences in protein expression patterns caused by transcriptional and post-transcriptional alterations related to insulin action and cellular lipid accumulation. PMID- 15131121 TI - Casein kinase 1alpha interacts with retinoid X receptor and interferes with agonist-induced apoptosis. AB - Agonists of retinoid X receptors (RXRs), which include the natural 9-cis-retinoic acid and synthetic analogs, are potent inducers of growth arrest and apoptosis in some cancer cells. As such, they are being used in clinical trials for the treatment and prevention of solid tumors and are used to treat cutaneous T cell lymphoma. However, the molecular mechanisms that underlie the anti-cancer effects of RXR agonists remain unclear. Here, we show that a novel pro-apoptotic pathway that is induced by RXR agonist is negatively regulated by casein kinase 1alpha (CK1alpha). CK1alpha associates with RXR in an agonist-dependent manner and phosphorylates RXR. The ability of an RXR agonist to recruit CK1alpha to a complex with RXR in cells correlates inversely with its ability to inhibit growth. Remarkably, depletion of CK1alpha in resistant cells renders them susceptible to RXR agonist-induced growth inhibition and apoptosis. Our study shows that CK1alpha can promote cell survival by interfering with RXR agonist induced apoptosis. Inhibition of CK1alpha may enhance the anti-cancer effects of RXR agonists. PMID- 15131122 TI - Evidence for cyclin D3 as a novel target of rapamycin in human T lymphocytes. AB - The immunosuppressant rapamycin has been shown to inhibit G(1)/S transition of the cell cycle. This inhibition is thought to be mediated by maintenance of the threshold levels of cyclin-dependent kinase (CDK) inhibitor p27(Kip1) (p27) and inhibition of p70 s6 kinase (p70(s6k)). However, recent evidence suggests that cells still remain sensitive to rapamycin in the absence of functional p27 or p70(s6k). Here, we show that rapamycin represses cyclin D3 levels in activated human T lymphocytes with no inhibitory effects on cyclin D2. Furthermore, rapamycin elicits similar cyclin D3 modulatory effects in B lymphocytes. The overall effect of rapamycin on cyclin D3 leads to impaired formation of active complexes with Cdk4 or Cdk6 and subsequent inhibition of cyclin D3/CDK kinase activity. Decrease in cyclin D3 protein levels is due to translational repression and not due to attenuated transcription of the cyclin D3 gene. Importantly, stable overexpression of cyclin D3 (2-2.5 fold) in Jurkat T cell transfectants renders them resistant to lower doses (1-10 ng/ml) of rapamycin. These results point to a critical role of cyclin D3 in rapamycin-mediated immunosuppressive effects in T cells and cell cycle regulation in lymphocytes in general. PMID- 15131123 TI - The oligomerization state determines regulatory properties and inhibitor sensitivity of type 4 cAMP-specific phosphodiesterases. AB - PDE4 splice variants are classified into long and short forms depending on the presence or absence of two unique N-terminal domains termed upstream conserved regions 1 and 2 (UCR1 and -2). We have shown previously that the UCR module mediates dimerization of PDE4 long forms, whereas short forms, which lack UCR1, behave as monomers. In the present study, we demonstrate that dimerization is an essential structural element that determines the regulatory properties and inhibitor sensitivities of PDE4 enzymes. Comparing the properties of the dimeric wild type PDE4D3 with several monomeric mutant PDE4D3 constructs revealed that disruption of dimerization ablates the activation of PDE4 long forms by either protein kinase A phosphorylation or phosphatidic acid binding. Moreover, the analysis of heterodimers consisting of a catalytically active and a catalytically inactive PDE4D3 subunit indicates that protein kinase A phosphorylation of both subunits is essential to fully activate PDE4 enzymes. In addition to affecting enzyme regulation, disruption of dimerization reduces the sensitivity of the enzymes toward the prototypical PDE4 inhibitor rolipram. Parallel binding assays indicated that this shift in rolipram sensitivity is likely mediated by a decrease in the number of inhibitor binding sites in the high affinity rolipram binding state. Thus, although dimerization is not a requirement for high affinity rolipram binding, it functions to stabilize PDE4 long forms in their high affinity rolipram binding conformation. Taken together, our data indicate that dimerization defines the properties of PDE4 enzymes and suggest a common structural and functional organization for all PDEs. PMID- 15131124 TI - MAGP-2 has multiple binding regions on fibrillins and has covalent periodic association with fibrillin-containing microfibrils. AB - The interactions of microfibril-associated glycoprotein (MAGP)-2 have been investigated with fibrillins and fibrillin-containing microfibrils. Solid phase binding assays were conducted with recombinant fragments covering fibrillin-1 and most of fibrillin-2. MAGP-2, and its structure relative MAGP-1, were found to bind two fragments spanning the N-terminal half of fibrillin-1 and an N-terminal fragment of fibrillin-2. Blocking experiments indicated that MAGP-2 had a binding site(s) close to the N terminus of the fibrillin-1 molecule that was distinct from that for MAGP-1 and an additional, more central binding site(s) that may be shared by the two MAGPs. Immunogold labeling of developing nuchal ligament tissue showed that MAGP-2 had regular covalent and periodic (about 56 nm) association with fibrillin-containing microfibrils of elastic fibers in this tissue. Further analysis of isolated microfibrils indicated that MAGP-2 was attached at two points along the microfibril substructure, "site 1" on the "beads" and "site 2" at the "shoulder" of the interbead region close to where the two "arms" fuse. In contrast, MAGP-1 was located only on the beads. Comparison of the MAGP-2 binding data with known fibrillin epitope maps of the microfibrils showed that site 1 correlated with the N-terminal MAGP-2 binding region, and site 2 correlated with the second, more central, MAGP-2 binding region on the fibrillin-1 molecule. Of particular note, immunolabeling at site 2 was markedly decreased, relative to that at site 1, on extended microfibrils with bead-to-bead periods over 90 nm, suggesting that site 2 may move toward the beads when the microfibril is stretched. The study points to MAGP-2 being an integral component of some populations of fibrillin-containing microfibrils. Moreover, the identification of multiple MAGP-binding sequences on fibrillins supports the concept that MAGPs may function as molecular cross-linkers, stabilizing fibrillin monomers in folded conformation within or between the microfibrils, and thus MAGPs may be implicated in the modulation of the elasticity of these structures. PMID- 15131125 TI - Mutants of plasminogen activator inhibitor-1 designed to inhibit neutrophil elastase and cathepsin G are more effective in vivo than their endogenous inhibitors. AB - Neutrophil elastase and cathepsin G are abundant intracellular neutrophil proteinases that have an important role in destroying ingested particles. However, when neutrophils degranulate, these proteinases are released and can cause irreparable damage by degrading host connective tissue proteins. Despite abundant endogenous inhibitors, these proteinases are protected from inhibition because of their ability to bind to anionic surfaces. Plasminogen activator inhibitor type-1 (PAI-1), which is not an inhibitor of these proteinases, possesses properties that could make it an effective inhibitor of neutrophil proteinases if its specificity could be redirected. PAI-1 efficiently inhibits surface-sequestered proteinases, and it efficiently mediates rapid cellular clearance of PAI-1-proteinase complexes. Therefore, we examined whether PAI-1 could be engineered to inhibit and clear neutrophil elastase and cathepsin G. By introducing specific mutations in the reactive center loop of wild-type PAI-1, we generated PAI-1 mutants that are effective inhibitors of both proteinases. Kinetic analysis shows that the inhibition of neutrophil proteinases by these PAI 1 mutants is not affected by the sequestration of neutrophil elastase and cathepsin G onto surfaces. In addition, complexes of these proteinases and PAI-1 mutants are endocytosed and degraded by lung epithelial cells more efficiently than either the neutrophil proteinases alone or in complex with their physiological inhibitors, alpha1-proteinase inhibitor and alpha1 antichymotrypsin. Finally, the PAI-1 mutants were more effective in reducing the neutrophil elastase and cathepsin G activities in an in vivo model of lung inflammation than were their physiological inhibitors. PMID- 15131126 TI - Ser/Arg-rich protein-mediated communication between U1 and U2 small nuclear ribonucleoprotein particles. AB - Previous work demonstrated that U1 small nuclear ribonucleoprotein particle (snRNP), bound to a downstream 5' splice site, can positively influence utilization of an upstream 3' splice site via exon definition in both trans- and cis-splicing systems. Although exon definition results in the enhancement of splicing of an upstream intron, the nature of the factors involved has remained elusive. We assayed the interaction of U1 snRNP as well as the positive effect of a downstream 5' splice site on trans-splicing in nematode extracts containing either inactive (early in development) or active (later in development) serine/arginine-rich splicing factors (SR proteins). We have determined that U1 snRNP interacts with the 5' splice site in the downstream exon even in the absence of active SR proteins. In addition, we determined that U1 snRNP-directed loading of U2 snRNP onto the branch site as well as efficient trans-splicing in these inactive extracts could be rescued upon the addition of active SR proteins. Identical results were obtained when we examined the interaction of U1 snRNP as well as the requirement for SR proteins in communication across a cis-spliced intron. Weakening of the 3' splice site uncovered distinct differences, however, in the ability of U1 snRNP to promote U2 addition, dependent upon its position relative to the branch site. These results demonstrate that SR proteins are required for communication between U1 and U2 snRNPs whether this interaction is across introns or exons. PMID- 15131127 TI - Galectin-12 is required for adipogenic signaling and adipocyte differentiation. AB - Galectin-12 is a member of the galectin family consisting of beta-galactoside binding proteins with conserved carbohydrate recognition domains. This protein is preferentially expressed in peripheral blood leukocytes and adipocytes. We previously showed that galectin-12 is induced by cell cycle block at the G(1) phase and causes G(1) arrest when overexpressed (Yang, R.-Y., Hsu, D. K., Yu, L., Ni, J., and Liu, F.-T. (2001) J. Biol. Chem. 276, 20252-20260). Here, we show that the galectin-12 gene is expressed in mouse preadipocytes and is up-regulated when preadipocytes undergo cell cycle arrest, concomitant with acquisition of the competence to undergo differentiation in response to adipogenic hormone stimulation. Following a brief down-regulation 1 day after adipogenic treatment, its expression was once again markedly elevated when cells underwent terminal differentiation. Down-regulation of endogenous galectin-12 expression by RNA interference greatly reduced the expression of the adipogenic transcription factors CCAAT/enhancer-binding protein-beta and -alpha and peroxisome proliferator-activated receptor-gamma and severely suppressed adipocyte differentiation as a result of defective adipogenic signaling. We conclude that galectin-12 is required for signal transduction that conveys hormone stimulation to the induction of adipogenic factors essential for adipocyte differentiation. The findings suggest that galectin-12 is a major regulator of adipose tissue development. PMID- 15131128 TI - Structure and function of recombinant cobra venom factor. AB - Cobra venom factor (CVF) is the complement-activating protein from cobra venom. It is a structural and functional analog of complement component C3. CVF functionally resembles C3b, the activated form of C3. Like C3b, CVF binds factor B, which is subsequently cleaved by factor D to form the bimolecular complex CVF,Bb. CVF,Bb is a C3/C5 convertase that cleaves both complement components C3 and C5. CVF is a three-chain protein that structurally resembles the C3b degradation product C3c, which is unable to form a C3/C5 convertase. Both C3 and CVF are synthesized as single-chain prepro-proteins. This study reports the recombinant expression of pro-CVF in two insect cell expression systems (baculovirus-infected Sf9 Spodoptera frugiperda cells and stably transfected S2 Drosophila melanogaster cells). In both expression systems pro-CVF is synthesized initially as a single-chain pro-CVF molecule that is subsequently proteolytically processed into a two-chain form of pro-CVF that structurally resembles C3. The C3 like form of pro-CVF can be further proteolytically processed into another two chain form of pro-CVF that structurally resembles C3b. Unexpectedly, all three forms of pro-CVF exhibit functional activity of mature, natural CVF. Recombinant pro-CVF supports the activation of factor B in the presence of factor D and Mg2+ and depletes serum complement activity like natural CVF. The bimolecular convertase pro-CVF,Bb exhibits both C3 cleaving and C5 cleaving activity. The activity of pro-CVF and the resulting C3/C5 convertase is indistinguishable from CVF and the CVF,Bb convertase. The ability to produce active forms of pro-CVF recombinantly ensures the continued availability of an important research reagent for complement depletion because cobra venom as the source for natural CVF will be increasingly difficult to obtain as the Indian cobra is on the list of endangered species. Experimental systems to express pro-CVF recombinantly will also be invaluable for studies to delineate the structure and function relationship of CVF and its differences from C3 as well as to generate human C3 derivatives with CVF-like function for therapeutic complement depletion ("humanized CVF"). PMID- 15131129 TI - Lysine beta311 of protein geranylgeranyltransferase type I partially replaces magnesium. AB - Protein geranylgeranyltransferase type I (GGTase I) catalyzes the attachment of a geranylgeranyl lipid group near the carboxyl terminus of protein substrates. Unlike protein farnesyltransferase (FTase) and protein geranylgeranyltransferase type II, which require both Zn(II) and Mg(II) for maximal turnover, GGTase I turnover is dependent only on Zn(II). In FTase, the magnesium ion is coordinated by aspartate beta352 and the diphosphate of farnesyl diphosphate to stabilize the developing charge in the transition state (Pickett, J. S., Bowers, K. E., and Fierke, C. A. (2003) J. Biol. Chem. 278, 51243-51250). In GGTase I, lysine beta311 is substituted for this aspartate and is proposed to replace the catalytic function of Mg(II) (Taylor, J. S., Reid, T. S., Terry, K. L., Casey, P. J., and Beese, L. S. (2003) EMBO J. 22, 5963-5974). Here we demonstrate that the prenylation rate constant catalyzed by wild type GGTase I (k(chem) = 0.18 +/- 0.02 s(-1)) is not dependent on Mg(II), is approximately 20-fold slower than the maximal rate constant catalyzed by FTase, and has a single pKa of 6.4 +/- 0.1, likely reflecting deprotonation of the peptide thiol. Mutation of lysine beta311 in GGTase I to alanine (Kbeta311A) or aspartate (Kbeta311D) decreases the k(chem) in the absence of magnesium 9-41-fold without significantly affecting the binding affinity of either substrate. Furthermore, the geranylgeranylation rate constant is enhanced by the addition of Mg(II) for Kbeta311A and Kbeta311D GGTase I 2-5 fold compared with wild type GGTase I with K(Mg) of 140 +/- 10 mm and 6.4 +/- 0.8 mm, respectively. These results demonstrate that lysine beta311 of GGTase I partially replaces the catalytic function of Mg(II) observed in FTase. PMID- 15131130 TI - Role of nuclear factor-kappaB in the antiviral action of interferon and interferon-regulated gene expression. AB - Interferons (IFNs) play critical roles in host defense by modulating the expression of various genes via tyrosine phosphorylation of STAT transcription factors. IFN-alpha/beta activates another important transcription factor, nuclear factor-kappaB (NF-kappaB), but its role in IFN-mediated activity is poorly understood. Sensitivity to the antiviral and gene-inducing effects of IFN was examined in normal fibroblasts and in NF-kappaB knockout fibroblasts from p50- and p65-null mice. Antiviral assays demonstrated that NF-kappaB knockout fibroblasts were sensitized to the antiviral action of IFN. Moreover, analysis of IFN-stimulated gene expression by real-time PCR demonstrated selective effects of NF-kappaB on gene expression. Our results demonstrate that a subset of IFN stimulated genes is regulated through an NF-kappaB-dependent pathway and that NF kappaB may regulate the sensitivity of cells to IFN-mediated antiviral activity. PMID- 15131131 TI - Strategic mutations in the class I major histocompatibility complex HLA-A2 independently affect both peptide binding and T cell receptor recognition. AB - Mutational studies of T cell receptor (TCR) contact residues on the surface of the human class I major histocompatibility complex (MHC) molecule HLA-A2 have identified a "functional hot spot" that comprises Arg(65) and Lys(66) and is involved in recognition by most peptide-specific HLA-A2-restricted TCRs. Although there is a significant amount of functional data on the effects of mutations at these positions, there is comparatively little biochemical information that could illuminate their mode of action. Here, we have used a combination of fluorescence anisotropy, functional assays, and Biacore binding experiments to examine the effects of mutations at these positions on the peptide-MHC interaction and TCR recognition. The results indicate that mutations at both position 65 and position 66 influence peptide binding by HLA-A2 to various extents. In particular, mutations at position 66 result in significantly increased peptide dissociation rates. However, these effects are independent of their effects on TCR recognition, and the Arg(65)-Lys(66) region thus represents a true "hot spot" for TCR recognition. We also made the observation that in vitro T cell reactivity does not scale with the half-life of the peptide-MHC complex, as is often assumed. Finally, position 66 is implicated in the "dual recognition" of both peptide and TCR, emphasizing the multiple roles of the class I MHC peptide binding domain. PMID- 15131132 TI - MINT, the Msx2 interacting nuclear matrix target, enhances Runx2-dependent activation of the osteocalcin fibroblast growth factor response element. AB - Msx2 promotes osteogenic lineage allocation from mesenchymal progenitors but inhibits terminal differentiation demarcated by osteocalcin (OC) gene expression. Msx2 inhibits OC expression by targeting the fibroblast growth factor responsive element (OCFRE), a 42-bp DNA domain in the OC gene bound by the Msx2 interacting nuclear target protein (MINT) and Runx2/Cbfa1. To better understand Msx2 regulation of the OCFRE, we have studied functional interactions between MINT and Runx2, a master regulator of osteoblast differentiation. In MC3T3E1 osteoblasts (with endogenous Runx2 and FGFR2), MINT augments transcription driven by the OCFRE that is further enhanced by FGF2 treatment. OCFRE regulation can be reconstituted in the naive CV1 fibroblast cell background. In CV1 cells, MINT synergizes with Runx2 to enhance OCFRE activity in the presence of activated FGFR2. The RNA recognition motif domain of MINT (which binds the OCFRE) is required. Runx2 structural studies reveal that synergy with MINT uniquely requires Runx2 activation domain 3. In confocal immunofluorescence microscopy, MINT adopts a reticular nuclear matrix distribution that overlaps transcriptionally active osteoblast chromatin, extensively co-localizing with the phosphorylated RNA polymerase II meshwork. MINT only partially co-localizes with Runx2; however, co-localization is enhanced 2.5-fold by FGF2 stimulation. Msx2 abrogates Runx2-MINT OCFRE activation, and MINT-directed RNA interference reduces endogenous OC expression. In chromatin immunoprecipitation assays, Msx2 selectively inhibits Runx2 binding to OC chromatin. Thus, MINT enhances Runx2 activation of multiprotein complexes assembled by the OCFRE. Msx2 targets this complex as a mechanism of transcriptional inhibition. In osteoblasts, MINT may serve as a nuclear matrix platform that organizes and integrates osteogenic transcriptional responses. PMID- 15131133 TI - Purification and characterization of the bacterial MraY translocase catalyzing the first membrane step of peptidoglycan biosynthesis. AB - The MraY translocase catalyzes the first membrane step of bacterial cell wall peptidoglycan synthesis (i.e. the transfer of the phospho-N-acetylmuramoyl pentapeptide motif onto the undecaprenyl phosphate carrier lipid), a reversible reaction yielding undecaprenylpyrophosphoryl-N-acetylmuramoyl-pentapeptide (lipid intermediate I). This essential integral membrane protein, which is considered as a very promising target for the search of new antibacterial compounds, has thus far been clearly underexploited due to its intrinsic refractory nature to overexpression and purification. We here report conditions for the high level overproduction and for the first time the purification to homogeneity of milligram quantities of MraY protein. The kinetic parameters and effects of pH, salts, cations, and detergents on enzyme activity are described, taking the Bacillus subtilis MraY translocase as a model. PMID- 15131134 TI - Pediatric adherence as a multidimensional and dynamic construct, involving a triadic partnership. AB - OBJECTIVE: The aim of this paper is to integrate and draw attention to research findings that support our conceptualization of adherence as being multidimensional, dynamic, and involving a triadic partnership. METHODS: A review of relevant articles found in Medline, PsychLit, and reference lists provided in pertinent articles was performed. RESULTS: Significant progress has been made in identifying disease-related and psychosocial correlates of pediatric adherence and in developing intervention programs to promote adherence. Both lines of work have proceeded without a unifying framework for conceptualizing and measuring pediatric adherence. We offer a model that views adherence as comprising three defining aspects: (a) multidimensional, in support of the complexity and interrelatedness of treatment components, (b) a triadic partnership, as in mutually influential exchanges within and among the caregiver-medical team, child medical team, and caregiver-child relationships, and (c) dynamic in relation to the notion of changes in developmental adaptive capacity, contextual characteristics, and disease course. CONCLUSIONS: Much work has yet to be done to validate, refine, and extend our model of adherence. Recommendations are offered for assessing each defining aspect, and a group-based methodology that accommodates longitudinal, prospective data is described. PMID- 15131135 TI - Child behavior problems and family functioning as predictors of adherence and glycemic control in economically disadvantaged children with type 1 diabetes: a prospective study. AB - OBJECTIVE: This prospective study examined how child behavior problems and family functioning predict adherence behavior and glucose regulation (glycemic control) in a sample of economically disadvantaged children. METHODS: Children with type 1 diabetes (N = 116; 58.6% African American) were assessed for externalizing and internalizing behavior problems and family adaptability and cohesion and followed for a mean of 3.8 years. Glycemic control (glycosylated hemoglobin [HbA1c]) was assessed at baseline and follow-up, and adherence was assessed at follow-up. RESULTS: Analyses controlled for baseline HbA1c and years to follow-up. Multivariate analyses indicated that better adherence was predicted by high family cohesion. Better glycemic control was predicted by high family cohesion, the absence of externalizing behavior problems, and the presence of internalizing behavior problems. In addition, tests of moderation indicated that better follow up glycemic control occurred among girls from high cohesion families and younger children from low adaptability families. Although better adherence predicted better glycemic control, adherence did not mediate the relationships of behavior problems or family functioning with glycemic control. CONCLUSIONS: A child's behavior problems and family functioning may influence both adherence to the diabetes regimen and glycemic control several years later, suggesting the potential value of interventions that address child behavior and family functioning. PMID- 15131136 TI - Transcranial Doppler ultrasonography and executive dysfunction in children with sickle cell disease. AB - OBJECTIVE: To identify behavioral manifestations of executive dysfunction that are associated with cerebrovasculopathy, as measured by transcranial Doppler (TCD) ultrasonography, in children with sickle cell disease (HbSS). METHODS: Participants were 62 children and adolescents with HbSS disease who had no documented history of cerebrovascular accident. Children were classified according to the National Institutes of Health Stroke Prevention Trial in Sickle Cell Anemia (STOP) criteria (i.e., normal, conditional, and abnormal). RESULTS: Although children with abnormal TCD values were rated by their parents as exhibiting greater executive dysfunction in the areas of inhibitory control, problem-solving flexibility, and modulation of emotional responses compared with children in the conditional group, these differences were not clinically significant. In contrast, teachers rated children in the abnormal TCD group as having clinically significant executive dysfunction as manifested in their ability to solve problems in working memory, plan and organize, and self-monitor. CONCLUSIONS: Our findings lend preliminary support for the utility of teacher completed screening instruments designed to assess everyday behaviors associated with executive dysfunction in children with HbSS disease, especially among those children at greatest risk for neurological impairment as identified by TCD ultrasonography. PMID- 15131137 TI - A review of empirically supported psychosocial interventions for pain and adherence outcomes in sickle cell disease. AB - OBJECTIVE: To review empirical studies of psychological interventions for pain and adherence outcomes among patients with sickle cell disease. METHOD: We conducted a literature review of studies using psychological interventions targeted at pain and/or adherence behaviors related to sickle cell disease. The American Psychological Association Division 12 Task Force criteria (Chambless criteria) were used to evaluate the empirical support for three categories of interventions (cognitive-behavioral techniques, interventions aimed at behavioral change, and social support interventions). RESULTS: A small number of intervention studies met criteria for demonstrating empirical efficacy. As a group, cognitive-behavioral techniques fall into the category of probably efficacious for sickle cell pain. Other intervention types were limited by inadequate research methodologies. CONCLUSIONS: Future studies will need to more stringently test outcomes related to acute crises (e.g., pain episodes) as well as day-to-day management of sickle cell disease to clarify the most efficacious intervention approaches. Implications and suggestions for future research directions are discussed. PMID- 15131138 TI - Posttraumatic stress disorder (PTSD) and posttraumatic stress symptoms (PTSS) in families of adolescent childhood cancer survivors. AB - OBJECTIVE: To describe rates and concordance of posttraumatic stress disorder (PTSD) and posttraumatic stress symptoms (PTSS) in adolescent childhood cancer survivors and their mothers and fathers. METHOD: Participants were 150 adolescent survivors of childhood cancer, 146 mothers, and 103 fathers who completed the Impact of Events Scale-Revised, the Posttraumatic Stress Disorder Reaction Index, and the PTSD module of the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, fourth edition. RESULTS: PTSS are common in families of childhood cancer survivors. Parents reported more symptomatology than former patients. Mothers and fathers had relatively equal rates of current PTSD and levels of PTSS. Nearly 30% of mothers met diagnostic criteria since their child's diagnosis, with 13.7% currently experiencing PTSD. Nearly 20% of families had at least one parent with current PTSD. Ninety-nine percent of the sample had at least one family member reexperiencing symptoms. CONCLUSIONS: Both PTSD and PTSS help in understanding the experience of adolescent cancer survivors and their families. Within families of childhood cancer survivors, it is likely that some member may be experiencing treatable bothersome memories, arousal, or avoidance specific to the cancer experience. PMID- 15131139 TI - Parenting adolescent girls with type 1 diabetes: parents' perspectives. AB - OBJECTIVE: To explore parents' perceptions of the impact of type 1 diabetes mellitus (DM) on the family and to better understand parent-child conflict, parental worries, and coping strategies related to the management of DM in adolescence. METHODS: Semistructured interviews were conducted with the parents of 30 adolescent girls who had DM for at least 1 year. Using standard procedures for content analysis, themes were identified from the parents' discussions. RESULTS: Although parents reported worries, conflicts, and negative impacts of the DM on their family, they also recognized positive aspects of the DM (e.g., it promotes adolescent responsibility) and had strategies to cope with its challenges (e.g., letting go of perfectionism in disease management, keeping a positive attitude). CONCLUSIONS: Findings from this study can be used by clinicians to assess parents' concerns about DM and to help them discover ways to cope with their worries and stay involved with the disease's management. Considering the parents' perspective will likely lead to parents feeling understood by the health care team and will help to reduce their worries. PMID- 15131140 TI - Psychometric properties of a new measure of fathers' involvement in the management of pediatric chronic diseases. AB - OBJECTIVE: Despite theoretical and empirical reasons suggesting its potential importance, paternal involvement in the management of pediatric chronic diseases has rarely been a primary focus of research on family adaptation to pediatric disease. This may be due to a lack of appropriate tools to measure relevant behaviors. This study assessed the reliability and validity of the Dads' Active Disease Support scale (DADS), which was designed to measure male and female caregivers' estimates of the amount and helpfulness of paternal involvement in managing six pediatric chronic diseases. METHOD: A sample of 224 heterosexual couples completed the DADS and measures of mother, child, and family functioning, yielding 190 who completed data sets. Of these, 91 mothers and 88 fathers completed the DADS again after a 1-month interval. RESULTS: Results confirmed DADS internal consistency, test-retest reliability, and interparent agreement. Confirmatory factor analysis supported its construct validity: A two-factor model (amount and helpfulness of fathers' involvement) best accounted for participants' responses. Significant correlations with scores on the Family Assessment Device supported DADS convergent validity. DADS scores of mothers and fathers suggest substantial room for improvement in both the amount and the helpfulness of paternal involvement in disease management. Mothers provided significantly higher ratings of the helpfulness of fathers' involvement than did fathers. CONCLUSIONS: The DADS appears to be a reliable and valid measure for studies of the associations between paternal involvement in disease management and child, maternal, and family adaptation to pediatric chronic medical conditions. PMID- 15131141 TI - Practical considerations in design of internal amplification controls for diagnostic PCR assays. PMID- 15131142 TI - Specific detection and identification of herpes B virus by a PCR-microplate hybridization assay. AB - Herpes B virus DNA was specifically amplified by PCR, targeting the regions that did not cross-react with herpes simplex virus (HSV). The amplified products, which were shown to be highly genetic polymorphisms among herpes B virus isolates, were identified by microplate hybridization with probes generated by PCR. The products immobilized in microplate wells were hybridized with the biotin labeled probes derived from the SMHV strain of herpes B virus. The amplified products derived from the SMHV and E2490 strains of herpes B virus were identified by microplate hybridization. PCR products amplified from the trigeminal ganglia of seropositive cynomolgus macaques were identified as herpes B virus DNA. The utility of the PCR-microplate hybridization assay for genetic detection and identification of the polymorphic region of herpes B virus was determined. PMID- 15131143 TI - New real-time PCR assay for rapid detection of methicillin-resistant Staphylococcus aureus directly from specimens containing a mixture of staphylococci. AB - Molecular methods for the rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) are generally based on the detection of an S. aureus specific gene target and the mecA gene. However, such methods cannot be applied for the direct detection of MRSA from nonsterile specimens such as nasal samples without the previous isolation, capture, or enrichment of MRSA because these samples often contain both coagulase-negative staphylococci (CoNS) and S. aureus, either of which can carry mecA. In this study, we describe a real-time multiplex PCR assay which allows the detection of MRSA directly from clinical specimens containing a mixture of staphylococci in <1 h. Five primers specific to the different staphylococcal cassette chromosome mec (SCCmec) right extremity sequences, including three new sequences, were used in combination with a primer and three molecular beacon probes specific to the S. aureus chromosomal orfX gene sequences located to the right of the SCCmec integration site. Of the 1,657 MRSA isolates tested, 1,636 (98.7%) were detected with the PCR assay, whereas 26 of 569 (4.6%) methicillin-susceptible S. aureus (MSSA) strains were misidentified as MRSA. None of the 62 nonstaphylococcal bacterial species or the 212 methicillin resistant or 74 methicillin-susceptible CoNS strains (MRCoNS and MSCoNS, respectively) were detected by the assay. The amplification of MRSA was not inhibited in the presence of high copy numbers of MSSA, MRCoNS, or MSCoNS. The analytical sensitivity of the PCR assay, as evaluated with MRSA-negative nasal specimens containing a mixture of MSSA, MRCoNS, and MSCoNS spiked with MRSA, was approximately 25 CFU per nasal sample. This real-time PCR assay represents a rapid and powerful method which can be used for the detection of MRSA directly from specimens containing a mixture of staphylococci. PMID- 15131144 TI - Evaluation of rapid diagnostic tests for typhoid fever. AB - Laboratory diagnosis of typhoid fever requires isolation and identification of Salmonella enterica serotype Typhi. In many areas where this disease is endemic, laboratory capability is limited. Recent advances in molecular immunology have led to the identification of sensitive and specific markers for typhoid fever and technology to manufacture practical and inexpensive kits for their rapid detection. We evaluated three commercial kits for serologic diagnosis of typhoid fever. Patients presenting with > or = 4 days of fever were enrolled at two hospitals in Southern Vietnam. Cases were patients with serotype Typhi isolated from blood samples, and controls were patients with other laboratory-confirmed illnesses. Serotype Typhi isolates were confirmed and tested for antimicrobial susceptibility at the Pasteur Institute in Ho Chi Minh City. The Widal test was run at the hospitals and the Pasteur Institute. Sera were shipped frozen to the Centers for Disease Control and Prevention and tested by using Multi-Test Dip-S Ticks, TyphiDot, and TUBEX to detect immunoglobulin G (IgG), IgG and IgM, and IgM, respectively. Package insert protocol instructions were followed. We enrolled 59 patients and 21 controls. The sensitivity and specificity findings were as follows: 89 and 53% for Multi-Test Dip-S-Ticks, 79 and 89% for TyphiDot, 78 and 89% for TUBEX, and 64 and 76% for Widal testing in hospitals and 61% and 100% for Widal testing at the Pasteur Institute. For all assays, the sensitivity was highest in the second week of illness. The Widal test was insensitive and displayed interoperator variability. Two rapid kits, TyphiDot and TUBEX, demonstrated promising results. PMID- 15131145 TI - Use of genome level-informed PCR as a new investigational approach for analysis of outbreak-associated Mycobacterium tuberculosis isolates. AB - Mycobacterium tuberculosis strain CH, the index isolate linked to a major tuberculosis outbreak associated with high levels of transmissibility and virulence, was characterized by microarray analysis by use of a PCR product array representative of the genome of M. tuberculosis strain H37Rv. Seven potential genomic deletions were identified in CH, five of which were confirmed by PCR analysis across the predicted deletion points. The panel of five PCRs required to individually interrogate these loci was collectively referred to as the genome level-informed PCR (GLIP) assay. GLIP analysis was performed with CH, 12 other epidemiologically linked isolates, and 43 recent, non-outbreak-associated isolates derived from patients within the local area. All 13 outbreak-linked isolates showed a profile corresponding to the presence of all five deletions. These 13 isolates were also found to share common variable-number tandem repeat and mycobacterial interspersed repetitive unit profiles. None of the 43 non outbreak-associated isolates exhibited the five-deletion profile. Although three individual deletions were present in upwards of 44% of the non-outbreak associated isolates, no single-deletion isolates were detected. Interestingly, none of these deletions had been previously recognized, and sequence analysis of the immediate flanking regions in CH failed to identify a likely mechanism of deletion for four of the five loci. The GLIP assay also proved valuable in ongoing surveillance of the outbreak, rapidly identifying a further two outbreak associated cases months after the initial cluster and, importantly, dismissing a further 12 epidemiologically suspect cases, which allowed the optimum deployment of public health resources. PMID- 15131146 TI - Distribution of insertion sequences associated with Tn1546-like elements among Enterococcus faecium isolates from patients in Korea. AB - The vanA gene cluster is carried as a part of Tn1546-like elements. The genetic diversity in Tn1546-like elements has been documented previously. The differences described thus far have included the integration of insertion sequence (IS) elements IS1216V, IS1251, IS1476, and IS1542. Among these, IS1216V has been reported to be widespread in VanA enterococci of diverse geographic areas, whereas IS1542 and IS1476 have been reported only in the United Kingdom and Canada, respectively. We investigated the distribution of ISs among 20 vanA containing Enterococcus faecium isolates from human patients in nine different university hospitals in Korea. Pulsed-field gel electrophoresis (PFGE) was performed to identify the clonality of the isolates. Moreover, PCR amplification of the internal regions of Tn1546 was performed for structural analysis of the van gene, and both DNA strands of the PCR amplicons were directly sequenced by the dideoxy termination method. The PFGE patterns revealed a high degree of clonal diversity. Structural analyses of the van gene detected IS1542 and IS1216V in the genomes of all 20 isolates, whereas it did not detect IS1476 or IS1251 in the genomes of any of the isolates. In addition, IS19 was detected in the vanS vanH intergenic region of one isolate. These data indicate that identification of the IS within a vanA gene cluster could be a useful tool in epidemiological investigations. In addition, the distribution of ISs associated with Tn1546-like elements among the Korean isolates is therefore similar to that among European vancomycin-resistant enterococci. PMID- 15131147 TI - Rapid screening method for detection of bacteria in platelet concentrates. AB - Public awareness has long focused on the risks of the transmission of viral agents through blood product transfusion. This risk, however, pales in comparison to the less publicized danger associated with the transfusion of blood products contaminated with bacteria, in particular, platelet concentrates. Up to 1,000 cases of clinical sepsis after the transfusion of platelet concentrates are reported annually in the United States. The condition is characterized by acute reaction symptoms and the rapid onset of septicemia and carries a 20 to 40% mortality rate. The urgent need for a method for the routine screening of platelet concentrates to improve patient safety has long been recognized. We describe the development of a rapid and highly sensitive method for screening for bacteria in platelet concentrates for transfusion. No culture period is required; and the entire procedure, from the time of sampling to the time that the final result is obtained, takes less than 90 min. The method involves three basic stages: the selective removal of platelets by filtration following activation with a monoclonal antibody, DNA-specific fluorescent labeling of bacteria, and concentration of the bacteria on a membrane surface for enumeration by solid phase cytometry. The method offers a universal means of detection of live, nondividing, or dead gram-negative and gram-positive bacteria in complex cellular blood products. The sensitivity is higher than those of the culture-based methods available at present, with a detection limit of 10 to 10(2) CFU/ml, depending upon the bacterial strain. PMID- 15131148 TI - Comparison of a duplex quantitative real-time PCR assay and the COBAS Amplicor CMV Monitor test for detection of cytomegalovirus. AB - A duplex quantitative real-time PCR (qPCR) assay was designed to detect both the polymerase gene (pol) and the glycoprotein gene (gB) of cytomegalovirus (CMV). The detection limit of the qPCR was determined to be 1 to 3 copies/reaction and the linear measure interval was 10(3) to 10(8) copies/ml. The qPCR system was compared to the COBAS Amplicor CMV Monitor test (COBAS) by an analysis of 138 plasma samples. Both systems detected CMV in 71 cases and had negative results for 33 samples. In addition, 34 samples were positive by qPCR and negative by the COBAS assay, but in no case was the COBAS result positive and the qPCR result negative. Thus, qPCR detected 48% more positive cases than the COBAS method. For samples with > or = 10(5) copies/ml by qPCR, a saturation effect was seen in the COBAS assay and quantification required dilution. Copy numbers for pol and gB by qPCR generally agreed. However, the reproducibility of qPCR assays and the need for an international standard are discussed. Discrepant copy numbers for pol and gB by qPCR were found for samples from two patients, and sequence analysis revealed that the corresponding CMV strains were mismatched at four nucleotide positions compared with the gB fragment primer sequences. In conclusion, a duplex qPCR assay in a real-time format facilitates quantitative measurements and minimizes the risk of false-negative results. PMID- 15131149 TI - Clinically feasible biofilm susceptibility assay for isolates of Pseudomonas aeruginosa from patients with cystic fibrosis. AB - Pseudomonas aeruginosa is the predominant cause of chronic airway infection in cystic fibrosis (CF). CF airway isolates are often tested for antibiotic susceptibility but are rarely eradicated by the antibiotics identified as potentially effective. The growth state of P. aeruginosa in CF airways is probably different from that exhibited under conventional susceptibility testing conditions and may represent a bacterial biofilm. Biofilm susceptibility testing methods were adapted to create an assay for implementation in a clinical microbiology laboratory. This assay gave reproducible results when examined in 300 paired determinations with 12 antimicrobial agents, with a serious error rate of 5.7%. The biofilm assay was used retrospectively to test these 12 agents against 94 isolates from 41 CF patients. The biofilm inhibitory concentrations (BICs) were much higher than the corresponding conventionally determined MICs for the beta-lactam antibiotics (median values: aztreonam, >128 microg/ml versus 4 microg/ml; ceftazidime, 128 microg/ml versus 2 microg/ml; piperacillin tazobactam, 256 microg/ml versus 4 microg/ml; and ticarcillin-clavulanate, 512 microg/ml versus 16 microg/ml, respectively) and doxycycline (>64 microg/ml versus 16 microg/ml); and similar for meropenem (4 micro g/ml versus < or = 1 microg/ml), ciprofloxacin (0.5 microg/ml versus 1 microg/ml), and the aminoglycosides amikacin (32 microg/ml versus 16 microg/ml), gentamicin (16 microg/ml versus 8 microg/ml), and tobramycin (4 microg/ml versus 2 microg/ml). The median BIC for azithromycin was 2 microg/ml, whereas isolates were uniformly resistant when tested by standard methods. This demonstrates the feasibility of adapting biofilm susceptibility methods to the clinical microbiology laboratory and opens the way to examining whether biofilm testing might be used to select more effective antibiotic combinations for CF airway infections than methods in current use. PMID- 15131150 TI - Sequencing and comparative analysis of flagellin genes fliC, fljB, and flpA from Salmonella. AB - Salmonella isolates have traditionally been classified by serotyping, the serologic identification of two surface antigens, O-polysaccharide and flagellin protein. Serotyping has been of great value in understanding the epidemiology of Salmonella and investigating disease outbreaks; however, production and quality control of the hundreds of antisera required for serotyping is difficult and time consuming. To circumvent the problems associated with antiserum production, we began the development of a system for determination of serotype in Salmonella based on DNA markers. To identify flagellar antigen-specific sequences, we sequenced 280 alleles of the three genes that are known to encode flagellin in Salmonella, fliC, fljB, and flpA, representing 67 flagellar antigen types. Analysis of the data indicated that the sequences from fliC, fljB, and flpA clustered by the antigen(s) they encode not by locus. The sequences grouped into four clusters based on their conserved regions. Three of the four clusters included multiple flagellar antigen types and were designated the G complex, the Z4 complex, and the alpha cluster. The fourth cluster contained a single antigen type, H:z(29). The amino acid sequences of the conserved regions within each cluster have greater than 95% amino acid identity, whereas the conserved regions differ substantially between clusters (75 to 85% identity). Substantial sequence heterogeneity existed between alleles encoding different flagellar antigens while alleles encoding the same flagellar antigen were homologous, suggesting that flagellin genes may be useful targets for the molecular determination of flagellar antigen type. PMID- 15131151 TI - Prevalence and characterization of a binary toxin (actin-specific ADP ribosyltransferase) from Clostridium difficile. AB - In addition to the two large clostridial cytotoxins (TcdA and TcdB), some strains of Clostridium difficile also produce an actin-specific ADP-ribosyltransferase, called binary toxin CDT. We used a PCR method and Southern blotting for the detection of genes encoding the enzymatic (CDTa) and binding (CDTb) components of the binary toxin in 369 strains isolated from patients with suspected C. difficile-associated diarrhea or colitis. Twenty-two strains (a prevalence of 6%) harbored both genes. When binary toxin production was assessed by Western blotting, 19 of the 22 strains reacted with antisera against the iota toxin of C. perfringens (anti-Ia and anti-Ib). Additionally, binary toxin activity, detected by the ADP-ribosyltransferase assay, was present in only 17 of the 22 strains. Subsequently, all 22 binary toxin-positive strains were tested for the production of toxins TcdA and TcdB, toxinotyped, and characterized by serogrouping, PCR ribotyping, arbitrarily primed PCR, and pulsed-field gel electrophoresis. All binary toxin-positive strains also produced TcdB and/or TcdA. However, they had significant changes in the tcdA and tcdB genes and belonged to variant toxinotypes III, IV, V, VII, IX, and XIII. We could differentiate 16 profiles by using typing methods, indicating that most of the binary toxin-positive strains were unrelated. PMID- 15131152 TI - Real-time PCR system for detection of orthopoxviruses and simultaneous identification of smallpox virus. AB - A screening assay for real-time LightCycler (Roche Applied Science, Mannheim, Germany) PCR identification of smallpox virus DNA was developed and compiled in a kit system under good manufacturing practice conditions with standardized reagents. In search of a sequence region unique to smallpox virus, the nucleotide sequence of the 14-kDa fusion protein gene of each of 14 variola virus isolates of the Russian World Health Organization smallpox virus repository was determined and compared to published sequences. PCR primers were designed to detect all Eurasian-African species of the genus ORTHOPOXVIRUS: A single nucleotide mismatch resulting in a unique amino acid substitution in smallpox virus was used to design a hybridization probe pair with a specific sensor probe that allows reliable differentiation of smallpox virus from other orthopoxviruses by melting curve analysis. The applicability was demonstrated by successful amplification of 120 strains belonging to the orthopoxvirus species variola, vaccinia, camelpox, mousepox, cowpox, and monkeypox virus. The melting temperatures (T(m)s) determined for 46 strains of variola virus (T(m)s, 55.9 to 57.8 degrees C) differed significantly (P = 0.005) from those obtained for 11 strains of vaccinia virus (T(m)s, 61.7 to 62.7 degrees C), 15 strains of monkeypox virus (T(m)s, 61.9 to 62.2 degrees C), 40 strains of cowpox virus (T(m)s, 61.3 to 63.7 degrees C), 8 strains of mousepox virus (T(m), 61.9 degrees C), and 8 strains of camelpox virus (T(m)s, 64.0 to 65.0 degrees C). As most of the smallpox virus samples were derived from infected cell cultures and tissues, smallpox virus DNA could be detected in a background of human DNA. By applying probit regression analysis, the analytical sensitivity was determined to be 4 copies of smallpox virus target DNA per sample. The DNAs of several human herpesviruses as well as poxviruses other than orthopoxviruses were not detected by this method. The assay proved to be a reliable technique for the detection of orthopoxviruses, with the advantage that it can simultaneously identify variola virus. PMID- 15131153 TI - mecA Locus diversity in methicillin-resistant Staphylococcus aureus isolates in Brisbane, Australia, and the development of a novel diagnostic procedure for the Western Samoan phage pattern clone. AB - An emerging public health phenomenon is the increasing incidence of methicillin resistant Staphylococcus aureus (MRSA) infections that are acquired outside of health care facilities. One lineage of community-acquired MRSA (CA-MRSA) is known as the Western Samoan phage pattern (WSPP) clone. The central aim of this study was to develop an efficient genotyping procedure for the identification of WSPP isolates. The approach taken was to make use of the highly variable region downstream of mecA in combination with a single nucleotide polymorphism (SNP) defined by the S. aureus multilocus sequence typing (MLST) database. The premise was that a combinatorial genotyping method that interrogated both a highly variable region and the genomic backbone would deliver a high degree of informative power relative to the number of genetic polymorphisms interrogated. Thirty-five MRSA isolates were used for this study, and their gene contents and order downstream of mecA were determined. The CA-MRSA isolates were found to contain a truncated mecA downstream region consisting of mecA-HVR-IS431 mec-dcs Ins117, and a PCR-based method for identifying this structure was developed. The hospital-acquired isolates were found to contain eight different mecA downstream regions, three of which were novel. The Minimum SNPs computer software program was used to mine the S. aureus MLST database, and the arcC 272G polymorph was identified as 82% discriminatory for ST-30. A real-time PCR assay was developed to interrogate this SNP. We found that the assay for the truncated mecA downstream region in combination with the interrogation of arcC position 272 provided an unambiguous identification of WSPP isolates. PMID- 15131154 TI - Development and evaluation of a novel loop-mediated isothermal amplification method for rapid detection of severe acute respiratory syndrome coronavirus. AB - The development and evaluation of a one-step single-tube accelerated real-time quantitative reverse transcription (RT) loop-mediated isothermal amplification (LAMP) assay is reported for rapid detection of the severe acute respiratory syndrome coronavirus (SARS-CoV) replicase gene. A total of 49 samples (15 throat washes, 13 throat swabs, and 21 combined throat and nasal swabs) collected from patients admitted to the Hanoi-French and Ninhbinh hospitals in Vietnam during the SARS epidemic were evaluated and compared to conventional RT-PCR. The RT-LAMP assay demonstrated 100-fold-greater sensitivity, with a detection limit of 0.01 PFU. The sensitivity and specificity of RT-LAMP assay for detecting viral RNA in clinical specimens with regard to RT-PCR were 100 and 87%, respectively. The specificity of the RT-LAMP assay was further validated by restriction analysis as well as nucleotide sequencing of the amplified product. The concentration of virus in most of the clinical samples was 0.1 PFU (0.1 to 10(2) PFU), as determined from the standard curve of SARS RT-LAMP and based on the time of positivity. The assay procedure is quite simple, wherein the amplification is carried out in a single tube under isothermal conditions at 63 degrees C, and the result can be obtained in less than 1 h (as early as 11 min). Thus, the RT-LAMP assay reported here has the advantages of rapid amplification, simple operation, and easy detection and will be useful for rapid and reliable clinical diagnosis of SARS-CoV in developing countries. PMID- 15131155 TI - Specific identification of Staphylococcus aureus by Staphychrom II, a rapid chromogenic staphylocoagulase test. AB - We compared the performance of Staphychrom II (International Microbio, Signes, France), a rapid (2-h) chromogenic staphylocoagulase test that uses human prothrombin and protease inhibitors, with those of the reference tube coagulase test (TCT) and the latex agglutination test (LAT) Slidex Staph Plus for the rapid identification of S. aureus. Prospective evaluation with 293 fresh clinical isolates yielded sensitivities, specificities, and predictive and negative predictive values of 98.1, 100, 100, and 95.1%, respectively, for the Staphychrom II test; 98.6, 98.7, 99.6, and 96.3%, respectively, for LAT; and 97.6, 98.7, 99.5, and 93.9%, respectively, for TCT. The perfect specificity of the Staphychrom II test was confirmed by testing 193 collection strains selected because of their potential testing pitfalls. The Staphychrom II test was positive for 90% of the 215 S. aureus strains tested after only 1 h of incubation. The Staphychrom II test was as sensitive as the reference TCT and was 100% specific. PMID- 15131156 TI - Performance assessment of DNA fragment sizing by high-sensitivity flow cytometry and pulsed-field gel electrophoresis. AB - The sizing of restriction fragments is the chief analytical technique utilized in the production of DNA fingerprints. Few techniques have been able to compete with pulsed-field gel electrophoresis (PFGE), which is capable of discriminating among bacteria at species and strain levels by resolving restriction fragments. However, an ultrasensitive flow cytometer (FCM) developed in our lab has also demonstrated the ability to discriminate bacteria at species and strain levels. The abilities of FCM warrant a quantitative parallel comparison with PFGE to assess and evaluate the accuracy and precision of DNA fragment sizing by both techniques. Replicate samples of Staphylococcus aureus Mu50 were analyzed along with two clinical S. aureus isolates. The absolute fragment sizing accuracy was determined for PFGE (5% +/- 2%) and FCM (4% +/- 4%), with sequence-predicted Mu50 SmaI fragment sizes used as a reference. Precision was determined by simple arithmetic methods (relative standard deviation for PFGE [RSD(PFGE) ] = 3% +/- 2% and RSD(FCM) = 1.2% +/- 0.8%) as well as by the use of dendrograms derived from Dice coefficient-unweighted pair group method with arithmetic averages (UPGMA) and Pearson-UPGMA analyses. All quantitative measures of PFGE and FCM precision were equivalent, within error. The precision of both methods was not limited by any single sample preparation or analysis step that was tracked in this study. Additionally, we determined that the curve-based clustering of fingerprint data provided a more informative and useful assessment than did traditional band-based methods. PMID- 15131157 TI - Variable ratio of hepatitis C virus RNA to viral core antigen in patient sera. AB - Quantification of hepatitis C virus (HCV) core antigen and RNA in serum samples leads to a highly variable ratio of both. It is not clear whether this is due to the inaccuracy of RNA quantification or whether both are independent parameters in a certain range. We established a real-time reverse transcription (RT)-PCR for HCV RNA that combines very high sensitivity with a large dynamic range and minimal standard deviations. The assay was calibrated with the first international standard, 96/790, and the international genotype panel for HCV from the National Institute of Biological Standardisation and Control. A linear readout was obtained between 200 and 5 x 10(7) IU/ml. The detection limit was 80 IU/ml, the reproducibility was <0.05 log, and the standard error within one run was <0.01. Comparison of the method with the Roche Monitor competitive RT-PCR revealed its high accuracy. The core protein concentration was determined within a range from 1.5 to 400 pg/ml by using the preliminary trak-C assay from Ortho Clinical Diagnostics. Correlating the HCV RNA levels with core antigen concentrations in 197 serum samples from 23 interferon-treated patients, a average ratio of 7,900 IU of HCV RNA per pg of core antigen was estimated, but the variability of this ratio exceeded largely the variability of the two assays, ranging from 50 to 20,000 IU/pg. Theoretically, HCV should contain ca. 43,000 IU of RNA/pg core. In conclusion, the core antigen assay seems to detect, in addition to complete virions, RNA-free core protein structures, which enhances its sensitivity (98% in this group). The variable ratio of RNA and core protein is not mainly due to standard deviations of quantification but could be an additional parameter for treatment follow-up and state of viral replication. PMID- 15131158 TI - Amplification of coccidioidal DNA in clinical specimens by PCR. AB - Coccidioides DNA was amplified from serum by a PCR using coccidioid-specific primers. A 239-bp product was visualized when 10 fg of exogenous coccidioidal DNA was subjected to amplification. This product was demonstrated in some human and mouse sera prior to the detection of coccidioidal antibodies. PMID- 15131160 TI - Reverse transcriptase PCR diagnostic assay for the coronavirus associated with severe acute respiratory syndrome. AB - Recent outbreaks of severe acute respiratory syndrome (SARS) have spurred intense research efforts around the world to deal with the serious threat to health posed by this novel coronavirus. A rapid, reliable diagnostic assay is needed for monitoring the spread of the disease. Here we report a method for eliminating false-negative results and increasing test sensitivity, based on the hypothesis that the message encoded by the nucleocapsid (N) gene is the most abundant during viral infection. Nasopharyngeal aspirates and stool samples were obtained from suspected SARS patients with major clinical symptoms and a significant history of close contact with infected patients. Total RNAs were extracted in a 96-well format, together with pig kidney epithelial (PK-15) cells as an internal control for extraction efficiency. PCR inhibitors were removed by ethanol precipitation, and a PCR for the pig beta-actin gene was used as a positive control for all clinical samples. Samples were analyzed by a reverse transcriptase PCR assay. Northern blot analysis was performed to demonstrate differences in subgenomic transcripts of the virus, and a real-time quantitative PCR was employed to compare the sensitivities of two loci (1b and N). The detection rate of the assay reached 44.4% on day 9 after the onset of the disease. The diagnostic PCR amplifying the N gene gave an average of a 26.0% (6.3 to 60.0%) stronger intensity signal than that for the 1b gene. In conclusion, the nucleocapsid gene represents an additional sensitive molecular marker for the diagnosis of the SARS coronavirus and can be further adapted for use in a high-throughput platform assay. PMID- 15131159 TI - Use of mycobacterial interspersed repetitive unit-variable-number tandem repeat typing to examine genetic diversity of Mycobacterium tuberculosis in Singapore. AB - Strain typing using variable-number tandem repeats of mycobacterial interspersed repetitive units (MIRU-VNTR) is a powerful tool for studying the epidemiology and genetic relationships of Mycobacterium tuberculosis isolates. For this study, isolates from 291 patients in Singapore were genotyped by this method. One hundred sixty-six distinct MIRU-VNTR patterns were detected. One hundred sixty two strains were grouped into 1 of 35 different MIRU-VNTR clusters and 131 isolates were unique. In this sample collection, 9 of the 12 MIRU-VNTR loci were moderately or highly discriminative according to their allelic diversities. The Hunter-Gaston discriminatory index was 0.975, indicating the high power of discrimination of MIRU-VNTR typing. By direct comparisons with previously typed MIRU-VNTR patterns and by genetic relationship analyses, we could identify and clearly define four epidemic groups of M. tuberculosis in our sample, corresponding to the W/Beijing, East-Africa-Indian, Haarlem, and Delhi genotype families. Furthermore, MIRU-VNTR typing was able to clearly distinguish ancestral and modern M. tuberculosis strains as defined by TbD1 genomic deletion analysis. These results indicate that MIRU-VNTR typing can be a useful first-line tool for studying the genetic diversity of M. tuberculosis isolates in a large urban setting such as Singapore. PMID- 15131161 TI - Assessment of virulence of pigeon isolates of Salmonella enterica subsp. enterica serovar typhimurium variant copenhagen for humans. AB - Salmonella enterica serovar Typhimurium variant Copenhagen was isolated from 5 of 152 (3.3%) feral pigeons from the city of Ghent (Belgium) and from 26 pooled fecal samples from 114 pigeon lofts (22.8%). These isolates belonged to phage type (PT) 99. Seven of the pigeon isolates were further compared in vitro to five human variant Copenhagen isolates, 2 isolates of PT 208, 1 isolate each of PT 120 and U302, and a nontypeable isolate. No differences in invasiveness in human intestinal epithelial Caco-2 cells were found. The human strains, however, were able to multiply significantly more inside human THP-1 macrophages than the pigeon strains. After inoculation of mice with a pigeon PT 99 strain, high numbers of Salmonella bacteria were shed with the feces, the internal organs were heavily colonized, and the animals showed severe clinical symptoms resulting in death. In conclusion, the less-pronounced ability of the pigeon variant Copenhagen strains to multiply inside human macrophages than human strains as well as the lack of human PT 99 isolates during 2002, despite the relatively high frequency of this PT in the pigeon population, suggest these strains to be of low virulence to humans. However, the high virulence for mice of the tested strain implies that rodents may act as reservoirs. PMID- 15131162 TI - Longitudinal study of the excretion patterns of thermophilic Campylobacter spp. in young pet dogs in Denmark. AB - The Campylobacter excretion patterns of 26 domestic pet dogs were described in a longitudinal study. The dogs entered the study between 3 and 8 months of age and were monitored until 2 years of age. They were tested monthly for Campylobacter carriage in stool samples that were cultured on the Campylobacter-selective media CAT and modified CCDA agar at 37 and 42 degrees C. This study comprised 366 fecal swab samples, of which 278 (76.2%) were found to be Campylobacter positive, with the following distribution of species: 75.0% Campylobacter upsaliensis, 19.4% Campylobacter jejuni, 2.1% Campylobacter lari, 0.7% Campylobacter coli, and 2.8% Campylobacter spp. Isolates were typed by pulsed-field gel electrophoresis (PFGE) to elucidate the strain excretion pattern. All study dogs excreted Campylobacter spp. during the study period. At 3 months of age, 60% of the dogs carried Campylobacter, increasing to nearly 100% carriers at 1 year of age, whereafter the carriage rate decreased to 67% at 24 months of age. The PFGE types showed that individual dogs were often colonized by unique strains of C. upsaliensis for several months, up to 21 months or longer. These C. upsaliensis strains were either clonal (or underwent concurrent minor mutative changes) or independent strains. In contrast, the excreted C. jejuni isolates were much more diverse and, in most cases, only seen in one sample from each dog. A high degree of diversity among different dogs was seen. We conclude that young domestic pet dogs excreted Campylobacter spp. during the majority of their puppyhood and adolescent period. In general C. upsaliensis strains were excreted for months, with short-term interruptions by or cocolonization with other transitory Campylobacter spp., predominantly C. jejuni. C. jejuni was more prevalent in dogs between 3 months and 1 year of age than in dogs between 1 and 2 years of age. PMID- 15131163 TI - Detection and differentiation of human parvovirus variants by commercial quantitative real-time PCR tests. AB - Parvovirus B19 causes a variety of diseases in humans, with outcomes ranging from asymptomatic to severe, such as chronic anemia in immunocompromised patients or fetal hydrops and death after maternal infection during pregnancy. The virus may be transmitted via plasma-derived products. According to the results of solvent detergent safety studies, an upper limit of B19 DNA in plasma pools was recently defined. To restrict the input of B19 virus into production pools, a quantitative nucleic acid test is a prerequisite. We examined the suitability of the two commercial quantitative B19 PCR tests, LightCycler-Parvovirus B19 quantification kit (Roche Diagnostics) and RealArt Parvo B19 LC PCR (Artus) for detection, quantification, and differentiation of the three known B19 genotypes, including the newly described erythrovirus variants (genotypes 2 and 3). The former kit was highly sensitive for genotype 1 but was not suitable for detection of genotype 2 or one of two genotype 3 strains. The latter kit detected and differentiated all three genotypes, albeit with lower sensitivity for one of the genotype-3 strains. We furthermore assessed the prevalence of the three B19 virus genotypes in blood donors, by screening pooled plasma samples derived from 140,160 Finnish blood donor units. None of the pools contained detectable levels of B19 virus genotypes 2 or 3. The origin, mode of transmission, and clinical significance of these genotypes are unknown and deserve further study. The RealArt Parvo B19 LC PCR is suitable for detection, quantification, and differentiation of all three B19 virus genotypes in molecular and clinical research. PMID- 15131164 TI - High seroprevalence of human T-cell lymphotropic virus type 1 in blood donors in Guyana and molecular and phylogenetic analysis of new strains in the Guyana shelf (Guyana, Suriname, and French Guiana). AB - The prevalence of human T-cell lymphotropic virus type 1 (HTLV-1) and HTLV-2 in blood donors in Guyana has never been estimated. We evaluated the prevalence of these viruses in blood donors by enzyme-linked immunosorbent assay and Western blotting and showed a prevalence of HTLV-1 of 1.3%; no HTLV-2 was detected. Female donors had a much higher HTLV-1 seroprevalence (3.6%) than male donors (0.7%). HTLV-1-seropositive donors tended to be slightly older than the average age for the total pool of donors. We also investigated the phylogenetic and molecular characteristics of HTLV-1 strains in Guyana and compared them with those identified in Suriname and French Guiana. Analysis of portions of the env and long terminal repeat nucleotide sequences showed that all the strains in Guyana and Suriname, like those in French Guiana, belonged to the transcontinental group of cosmopolitan subtype A. The similarities were greater between strains from Suriname and Guyana than between strains from Suriname and Guyana and those from French Guiana. Nevertheless, our results confirm that the HTLV-1 strains in all three countries have a common African origin. PMID- 15131165 TI - Expertise of French laboratories in detection, genotyping, and quantification of hepatitis C virus RNA in serum. AB - Before initiating new large-scale therapeutic trials for hepatitis C virus (HCV) infected patients, the French Health Authorities for HCV research decided to organize an evaluation of the expertise of laboratories that could be engaged to undertake molecular biology assays in such trials; 21 experienced laboratories participated in this national evaluation of laboratory expertise, which was performed in two successive rounds. The first round evaluated the laboratories for their abilities to detect HCV RNA in serum, determine genotypes, and quantify HCV RNA loads. The results observed by qualitative assays for HCV RNA detection were 100% sensitivity and 100% specificity for all laboratories. The genotyping results were 100% concordant for 9 laboratories and greater than 90% for 10 laboratories. By contrast, large coefficients of variation were observed for quantitative determination of HCV RNA loads, leading to a second round with standardized quantitative assays only. The dispersion of the results was larger by the AMPLICOR HCV Monitor assay than by the branched-DNA assay (mean coefficients of variation, 57.4 and 16.9%, respectively). In the majority of cases, discrepancies between the results of the two tests were found for samples with high viral loads. These results indicate the usefulness of validating, by controlling for expertise, both the reliabilities of laboratories involved in multicenter work and the standardized assays chosen for use in the evaluation of the biological impacts of new therapies. PMID- 15131166 TI - Detection of Shigella by a PCR assay targeting the ipaH gene suggests increased prevalence of shigellosis in Nha Trang, Vietnam. AB - Shigella spp. are exquisitely fastidious gram-negative organisms which frequently escape detection by traditional culture methods. To get a more complete understanding of the disease burden caused by Shigella in Nha Trang, Vietnam, real-time PCR was used to detect Shigella DNA. Randomly selected rectal swab specimens from 60 Shigella culture-positive patients and 500 Shigella culture negative patients detected by population-based surveillance of patients seeking care for diarrhea were processed by real-time PCR. The target of the primer pair is the invasion plasmid antigen H gene sequence (ipaH), carried by all four Shigella species and enteroinvasive Escherichia coli. Shigella spp. could be isolated from the rectal swabs of 547 of 19,206 (3%) patients with diarrhea. IpaH was detected in 55 of 60 (93%) Shigella culture-positive specimens, whereas it was detected in 87 of 245 (36%) culture-negative patients free of dysentery (P < 0.001). The number of PCR cycles required to detect a PCR product was highest for culture-negative, nonbloody diarrheal specimens (mean number of cycles to detection, 36.6) and was lowest for children with culture-positive, bloody diarrheal specimens (mean number of cycles, 25.3) (P < 0.001). The data from real time PCR amplification indicate that the culture-proven prevalence of Shigella among patients with diarrhea may underestimate the prevalence of Shigella infections. The clinical presentation of shigellosis may be directly related to the bacterial load. PMID- 15131167 TI - Clonal diversity and stability of subgingival Eikenella corrodens. AB - Eikenella corrodens is a commensal subgingival bacterium commonly found in both periodontally nondiseased and diseased subjects. The present study examined the clonal diversity and stability of subgingival E. corrodens over time. Ninety-five subjects were enrolled at the baseline examination, including 44 periodontally nondiseased subjects and 51 subjects with aggressive periodontitis. Twenty-two nondiseased subjects and 11 subjects with aggressive periodontitis were subsequently reexamined after an average interval of 14 months. Two subgingival plaque samples were obtained from each subject to determine the total cultivable bacteria. In addition, multiple E. corrodens isolates from each sample were recovered for clonal analysis by arbitrarily primed PCR. The mean numbers of distinct E. corrodens clones harbored by nondiseased subjects and subjects with aggressive periodontitis were 1.3 and 3.0, respectively. Thirty-nine percent of the nondiseased subjects and 63% of the subjects with aggressive periodontitis harbored multiple clones of E. corrodens. The numbers of distinct E. corrodens clones increased significantly (Mann-Whitney ranking test, P < 0.05) in sites from patients with aggressive periodontitis, in sites with pocket depths of 4 mm or greater, in sites with a clinical attachment loss of 2 mm or greater, and in sites coinfected with Porphyromonas gingivalis. Comparison of E. corrodens clones recovered at the baseline and those recovered at the follow-up examination showed that E. corrodens colonization was not stable. Thirty-eight of the 66 follow-up samples (58%) showed a complete change (including de novo colonization of the sites or complete elimination of the organism from the sites) of the subgingival E. corrodens clonal types between the baseline and the follow-up examinations. Our results suggest a complexity of subgingival microbiota not seen previously. PMID- 15131168 TI - Evaluation of advanced reverse transcription-PCR assays and an alternative PCR target region for detection of severe acute respiratory syndrome-associated coronavirus. AB - First-generation reverse transcription-PCR (RT-PCR) assays for severe acute respiratory syndrome-associated coronavirus (SARS-CoV) gave false-negative results in a considerable fraction of patients. In the present study, we evaluated two second-generation, replicase (R) gene-based, real-time RT-PCR test kits--the RealArt HPA coronavirus LC kit (Artus, Hamburg, Germany) and the LightCycler SARS-CoV quantification kit (Roche, Penzberg, Germany)--and a real time RT-PCR assay for the nucleocapsid (N) gene. Detecting the N-gene RNA might be advantageous due to its high abundance in cells. The kits achieved sensitivities of 70.8% (Artus) and 67.1% (Roche) in 66 specimens from patients with confirmed SARS (samples primarily from the upper and lower respiratory tract and stool). The sensitivity of the N-gene assay was 74.2%. The differences in all of the sensitivities were not statistically significant (P = 0.680 [analysis of variance]). Culture cells initially contained five times more N- than R-gene RNA, but the respective levels converged during 4 days of virus replication. In clinical samples the median concentrations of R- and N-gene RNA, respectively, were 1.2 x 10(6) and 2.8 x 10(6) copies/ml (sputum and endotracheal aspirates), 4.3 x 10(4) and 5.5 x 10(4) copies/ml (stool), and 5.5 x 10(2) and 5.2 x 10(2) copies/sample (throat swabs and saliva). Differences between the samples types were significant but not between the types of target RNA. All (n = 12) samples from the lower respiratory tract tested positive in all tests. In conclusion, the novel assays are more sensitive than the first-generation tests, but they still do not allow a comprehensive ruling out of SARS. Methods for the routine sampling of sputum without infection risk are needed to improve SARS RT-PCR. PMID- 15131169 TI - Genetic variability and molecular epidemiology of respiratory syncytial virus subgroup a strains in Japan determined by heteroduplex mobility assay. AB - We used heteroduplex mobility assay (HMA) to determine the genetic variability of 118 respiratory syncytial virus (RSV) field isolates from 19 epidemics occurring in a Japanese urban area between 1980 and 2000. Nucleotides 1 to 584 of the attachment G glycoprotein gene were amplified by reverse transcription-PCR, and the PCR amplicons were analyzed by HMA by using the earliest isolate from 1980 as the reference throughout. We also performed PCR-restriction fragment length polymorphism (RFLP) analysis and phylogenetic analysis on the same nucleotide sequence. PCR-RFLP revealed 9 patterns, whereas HMA produced 31 distinct patterns. The RFLP patterns were divided into two to seven distinct HMA genotypes. Field strains with similar degrees of G gene nucleotide differences from the reference strain often showed distinct HMA types. The RSV genetic heterogeneity detected by direct sequencing of the PCR amplicon was usually identical to HMA analysis. Analysis of the molecular epidemiology of RSV subgroup A isolates obtained by HMA showed that new RSV variants emerged with each epidemic and that previously dominant variants seldom recurred in subsequent epidemics. HMA is useful in detecting genetic variants of RSV subgroup A and has some advantages over other conventional methods. PMID- 15131170 TI - DNA macroarray for identification and typing of Staphylococcus aureus isolates. AB - A DNA macroarray containing 465 intragenic amplicons was designed to identify Staphylococcus aureus at the species level and to type S. aureus isolates. The genes selected included those encoding (i) S. aureus-specific proteins, (ii) staphylococcal and enterococcal proteins mediating antibiotic resistance and factors involved in their expression, (iii) putative virulence proteins and factors controlling their expression, and (iv) proteins produced by mobile elements. The macroarray was hybridized with the cellular DNAs of 80 S. aureus clinical isolates that were previously typed by analyses of their antibiograms and SmaI patterns. The set selected contained unrelated, endemic, and outbreak related isolates belonging to 45 SmaI genotypes. In a gene content dendrogram, the 80 isolates were distributed into 52 clusters. The outbreak-related isolates were linked in the same or a closely related cluster(s). Clustering based on gene content provided a better discrimination than SmaI pattern analysis for the tested mecA(+) isolates that were endemic to Europe. All of the antibiotic resistance genes detected could be correlated with their corresponding phenotypes, except for one isolate which carried a mecA gene without being resistant. The 16 isolates responsible for bone infections were distinguishable from the 12 isolates from uninfected nasal carriers by a significantly higher prevalence of the sdrD gene coding for a putative SD (serine-aspartate) adhesin (in 15 and 7 isolates, respectively). In conclusion, the macroarray designed for this study offers an attractive and rapid typing method which has the advantage of providing additional information concerning the gene content of the isolate of interest. PMID- 15131171 TI - Comparison of conventional and molecular methods for identification of aerobic catalase-negative gram-positive cocci in the clinical laboratory. AB - Over a period of 18 months we have evaluated the use of 16S ribosomal DNA (rDNA) sequence analysis as a means of identifying aerobic catalase-negative gram positive cocci in the clinical laboratory. A total of 171 clinically relevant strains were studied. The results of molecular analyses were compared with those obtained with a commercially available phenotypic identification system (API 20 Strep system; bioMerieux sa, Marcy l'Etoile, France). Phenotypic characterization identified 67 (39%) isolates to the species level and 32 (19%) to the genus level. Seventy-two (42%) isolates could not be discriminated at any taxonomic level. In comparison, 16S rDNA sequencing identified 138 (81%) isolates to the species level and 33 (19%) to the genus level. For 42 of 67 isolates assigned to a species with the API 20 Strep system, molecular analyses yielded discrepant results. Upon further analysis it was concluded that among the 42 isolates with discrepant results, 16S rDNA sequencing was correct for 32 isolates, the phenotypic identification was correct for 2 isolates, and the results for 8 isolates remained unresolved. We conclude that 16S rDNA sequencing is an effective means for the identification of aerobic catalase-negative gram-positive cocci. With the exception of Streptococcus pneumoniae and beta-hemolytic streptococci, we propose the use of 16S rDNA sequence analysis if adequate species identification is of concern. PMID- 15131172 TI - PCR-based assay for differentiation of Pseudomonas aeruginosa from other Pseudomonas species recovered from cystic fibrosis patients. AB - Pseudomonas aeruginosa is the major opportunistic bacterial pathogen in persons with cystic fibrosis (CF); pulmonary infection occurs in approximately 80% of adult CF patients. Much of CF patient management depends on accurate identification of P. aeruginosa from sputum culture. However, identification of this species may be problematic due to the marked phenotypic variability demonstrated by CF sputum isolates and the presence of other closely related species. To facilitate species identification, we used 16S ribosomal DNA (rDNA) sequence data to design PCR assays intended to provide genus- or species-level identification. Both assays yielded DNA fragments of the predicted size. We tested 42 culture collection strains (including 14 P. aeruginosa strains and 28 strains representing 16 other closely related Pseudomonas species) and 43 strains that had been previously identified as belonging to 28 nonpseudomonal species also recovered from CF patient sputum. Based on these 85 strains, the specificity and sensitivity of both assays were 100%. To further assess the utility of the PCR assays, we tested 66 recent CF sputum isolates. The results indicated that preliminary phenotypic testing had misidentified several isolates. The 16S rDNA sequence was determined for 38 isolates, and in all cases it confirmed the results of the PCR assays. Thus, we have designed two PCR assays: one is specific for the genus Pseudomonas, while the other is specific for P. aeruginosa. Both assays show 100% sensitivity and specificity. PMID- 15131173 TI - Widespread skin and soft-tissue infections due to two methicillin-resistant Staphylococcus aureus strains harboring the genes for Panton-Valentine leucocidin. AB - Infections caused by community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) are emerging as a major public health problem. CA-MRSA has been associated previously with skin and soft-tissue infection (SSTI) and with carriage of staphylococcal cassette chromosome mec (SCCmec) type IV and the Panton-Valentine leucocidin (PVL) virulence factor. To assess the clonal distribution of PVL-carrying strains and the association with SSTI in the San Francisco Bay area, we surveyed six collections of S. aureus isolates-671 isolates in all-collected between 1997 and 2002 originating from inpatient and outpatient clinical specimens and from a community-based sampling. Isolates were genotyped by pulsed-field gel electrophoresis, multilocus restriction fragment typing, and multilocus sequence typing and assayed for the PVL virulence factor. The S. aureus populations showed a high proportion of PVL-carrying strains, with frequencies ranging up to 70% in MRSA isolated from jail inmate patients and 69% in MRSA from patients receiving surgical treatment at an outpatient clinic specializing in treating SSTIs. PVL-carrying isolates were identified in nine clonal groups, but 88.5% of the PVL-carrying MRSA isolates belonged to only two clonal groups. These two clonal groups carried the SCCmec type IV resistance determinant and were more likely than other clonal groups to be recovered from SSTI sites than from other sites (P < 0.0001). There is evidence of clonal replacement over the period from 1999 to 2002, with one of these two clonal groups being supplanted by the other. PMID- 15131174 TI - Rapid quantification of drug resistance gene expression in Candida albicans by reverse transcriptase LightCycler PCR and fluorescent probe hybridization. AB - We developed a rapid, sensitive, and reproducible assay to quantify Candida albicans ACT1, CDR1, CDR2, ERG11, and MDR1 mRNA using a two-step reverse transcription and LightCycler real-time PCR (RT-LightCycler PCR) method with sequence-specific hybridization probes. We compared RT-LightCycler PCR with Northern hybridization for quantitative analysis of gene expression in isolates with various fluconazole susceptibilities. Specificity of each LightCycler PCR was verified by LightCycler melting curve analysis and agarose gel electrophoresis of amplified products. Correlation of quantification results between RT-LightCycler PCR and Northern hybridization yielded correlation coefficients of > or = 0.91 for all genes except MDR1 (0.74). In this case, reduced correlation was due to the inability of Northern hybridization to accurately quantify the high MDR1 expression in a susceptible dose-dependent isolate which was shown by RT-LightCycler PCR to overexpress MDR1 >200-fold relative to the other isolates tested. In four isolates, low levels of CDR2 mRNA were detected by RT-LightCycler PCR but were undetectable by Northern hybridization. mRNA quantification by RT-LightCycler PCR correlates with Northern hybridization and offers additional advantages, including increased sensitivity and speed of analysis, along with lower RNA concentration requirements and an increased dynamic range of signal detection. PMID- 15131175 TI - Comparison of two real-time quantitative assays for detection of severe acute respiratory syndrome coronavirus. AB - The new severe acute respiratory syndrome (SARS) coronavirus (CoV), described in February 2003, infected a total of 8,439 people. A total of 812 people died due to respiratory insufficiency. Close contact with symptomatic patients appeared to be the main route of transmission. However, potential transmission by blood transfusion could not be definitely excluded. Two real-time SARS-specific PCR assays were assessed for their sensitivities, agreement of test results, and intra-assay variabilities. Both assays rely on reverse transcription and amplification of extracted RNA. Dilutions of gamma-irradiated cell culture supernatants of SARS CoV-infected Vero E6 cells were prepared to determine the precisions, linear ranges, and accuracies of the assays. The linear range for the Artus RealArt HPA-Coronavirus assay (Artus assay) was 1 x 10(2) to 1 x 10(7) copies/ml, and that for the Roche LightCycler SARS CoV Quantification kit (Roche assay) was 1 x 10(4) to 2 x 10(8) copies/ml. The detection limit of the Roche assay was 3,982.1 copies/ml, whereas that of the Artus assay was 37.8 copies/ml. Detection limits were calculated with a standard preparation that was recommended for use by the World Health Organization. However, quantification of CoV in this preparation may be imprecise. In summary, both assays are suitable for quantitative measurement of SARS CoV at the high concentrations expected in sputum samples. The Artus assay is also suitable for detection of SARS CoV at the low concentrations found in serum samples. PMID- 15131176 TI - Detection of Candida in concentrated oral rinse cultures by real-time PCR. AB - The incidence of oral candidosis has increased in recent years, largely as a result of the emergence of human immunodeficiency virus infection and the more widespread use of immunosuppressive chemotherapy. This development has been associated with a need for more reliable methods for the detection of Candida. The present study assessed the performance of a real-time PCR and two block-based PCRs for the detection of Candida in 193 concentrated oral rinse culture (CRC) specimens. A total of 102 CRC specimens were positive by culture for Candida; and 96, 90, and 75 of these were also positive by real-time, N18-specific, and internal transcribed spacer (ITS)-specific PCRs, respectively. The five false negative results by the real-time PCR were all non-Candida albicans positive by culture. Of the 91 culture-negative CRC specimens, 20, 41, and 44 were positive by the real-time PCR and the N18- and ITS-specific PCRs, respectively. All three PCRs detected fungal DNA in 8 culture-negative CRC specimens, with a further 30 being positive by two of the three PCRs. A total of 32 CRC specimens were Candida free by all methods. In summary, a real-time PCR that provides a sensitive, specific, and rapid alternative technique for detection of Candida in the mouth is described. PMID- 15131177 TI - Comparison of two 3ABC enzyme-linked immunosorbent assays for diagnosis of multiple-serotype foot-and-mouth disease in a cattle population in an area of endemicity. AB - The development of a serological test for foot-and-mouth disease virus (FMDV) which is quick and easy to use, which can identify all seven serotypes, and which can differentiate vaccinated from convalescing or potential virus carriers would be a major advance in the epidemiological toolkit for FMDV. The nonstructural polyprotein 3ABC has recently been proposed as such an antigen, and a number of diagnostic tests are being developed. This paper evaluates the performance of two FMDV tests for antibodies to nonstructural proteins in an unvaccinated cattle population from a region of Cameroon with endemic multiple-serotype FMD. The CHEKIT-FMD-3ABC bo-ov (CHEKIT) enzyme-linked immunosorbent assay (ELISA) (Bommeli Diagnostics/Intervet) is a commercially available test that was compared with a competitive 3ABC ELISA (C-ELISA) developed in Denmark. The tests were compared with the virus neutralization test as the "gold standard." Diagnostic sensitivity and specificity were examined over a range of test cutoffs by using receiver operating characteristic curves, which allowed comparison of the overall performance of each test. The results indicated that the CHEKIT ELISA kit was 23% sensitive and 98% specific and the Danish C-ELISA was 71% sensitive and 90% specific at the recommended cutoff. These results have important implications if the tests are to be used to screen herds or individual cattle in surveillance programs, at border crossings for import-export clearance, or following emergency vaccination in an outbreak situation. PMID- 15131178 TI - Development of a bacteriophage phage replication assay for diagnosis of pulmonary tuberculosis. AB - Successful infection and replication of bacteriophages is indicative of the presence of viable bacteria. We describe here the development of a bacteriophage replication assay for the detection of Mycobacterium tuberculosis by using mycobacteriophage D29. Optimization of phage inoculate and incubation times allowed highly sensitive detection of M. bovis BCG. Fewer than 10 CFU (100 CFU/ml) were detected. No false-positive results were observed in negative samples. Application of the assay to 496 sputum specimens in the National Reference Laboratory of Zambia produced sensitivity, specificity, and positive and negative predictive values of 44.1, 92.6, 82.2, and 67.5%, respectively, compared to culture on Lowenstein-Jensen medium. The equivalent corresponding results for direct fluorescent smear microscopy were 42.3, 96.8, 91.2, and 67.6%. The small increase in sensitivity over that of direct microscopy does not justify the introduction of this technique for routine diagnosis of pulmonary tuberculosis at this time. PMID- 15131179 TI - Serotyping of 800 strains of Erysipelothrix isolated from pigs affected with erysipelas and discrimination of attenuated live vaccine strain by genotyping. AB - Eight hundred Erysipelothrix strains isolated between 1992 and 2002 from swine with erysipelas in Japan were serotyped. Thirty-seven, 47, 73, and 643 strains were isolated from animals with acute septicemia, urticaria, chronic endocarditis, and chronic arthritis, respectively, of which 381, 146, 254, and 19 isolates belonged to serotypes 1a, 1b, and 2b and other serotypes, respectively. All serotype 1a isolates were further examined for acriflavine resistance and their genotypes to discriminate them from the attenuated live vaccine strain, defined as serotype 1a, which is resistant to 0.02% acriflavine and which shows low levels of pathogenicity in mice. Of the serotype 1a isolates, 64.6% were acriflavine resistant, with 98.4% of these acriflavine-resistant strains having been isolated from animals with chronic arthritis. By randomly amplified polymorphic DNA (RAPD) analysis, almost all the acriflavine-resistant serotype 1a strains showed the 253-bp band characteristic of vaccine strains and were easily discriminated from all 113 strains of acriflavine-sensitive serotype 1a strains from animals with acute and subacute swine erysipelas. The incidence of acriflavine-resistant strains of the distinctive RAPD type 1-2 was markedly higher than that of the other RAPD types and serotypes. RAPD type 1-2 strains also included a specific group identifiable by restriction fragment length polymorphism DNA analysis. Furthermore, the pathogenicities of 29 isolates of RAPD type 1-2 for mice were lower than those of the 21 isolates of other RAPD types. Our results indicate that RAPD type 1-2 strains are live vaccine strains and that 37% of the cases of chronic swine erysipelas detected in the past 11 years in Japan have occurred as a side effect of live vaccine use. PMID- 15131180 TI - Incidence of group C human rotavirus in central Australia and sequence variation of the VP7 and VP4 genes. AB - Human group C rotavirus was identified in central Australia in each of eight years over a 16-year period between 1982 and 1997. Cases occurred either sporadically but over a relatively short period of time or as clustered outbreaks. These are the only reports of human group C rotavirus in Australia other than that of a single case reported approximately 1,800 km away in 1982. The electrophoretic genome profiles of isolates were identical for all those identified within the same year but different between those identified in different years. The VP7 genes of four isolates identified in four different years over a 7-year period between 1987 and 1993, and the VP4 genes of two of these isolates showed relatively little variation in genome and deduced amino acid sequence upon comparison of the equivalent genes between isolates. The sequences were also very similar to those from the corresponding genes from most of the human group C rotavirus isolates from other countries. This continues the observation of a high degree of gene sequence conservation among human group C rotaviruses worldwide. PMID- 15131181 TI - Simultaneous analysis of multiple staphylococcal enterotoxin genes by an oligonucleotide microarray assay. AB - Staphylococcal enterotoxins (SEs) are a family of 17 major serological types of heat-stable enterotoxins that are one of the leading causes of gastroenteritis resulting from consumption of contaminated food. SEs are considered potential bioweapons. Many Staphylococcus aureus isolates contain multiple SEs. Because of the large number of SEs, serological typing and PCR typing are laborious and time consuming. Furthermore, serological typing may not always be practical because of antigenic similarities among enterotoxins. We report on a microarray-based one tube assay for the simultaneous detection and identification (genetic typing) of multiple enterotoxin (ent) genes. The proposed typing method is based on PCR amplification of the target region of the ent genes with degenerate primers, followed by characterization of the PCR products by microchip hybridization with oligonucleotide probes specific for each ent gene. We verified the performance of this method by using several other techniques, including PCR amplification with gene-specific primers, followed by gel electrophoresis or microarray hybridization, and sequencing of the enterotoxin genes. The assay was evaluated by analysis of previously characterized staphylococcal isolates containing 16 ent genes. The microarray assay revealed that some of these isolates contained additional previously undetected ent genes. The use of degenerate primers allows the simultaneous amplification and identification of as many as nine different ent genes in one S. aureus strain. The results of this study demonstrate the usefulness of the oligonucleotide microarray assay for the analysis of multitoxigenic strains, which are common among S. aureus strains, and for the analysis of microbial pathogens in general. PMID- 15131182 TI - Evaluation of "Helicobacter heilmannii" subtypes in the gastric mucosas of cats and dogs. AB - Infection with candidatus "Helicobacter heilmannii" is associated with gastritis and mucosa-associated lymphoid tissue lymphoma in people. Infection with "H. heilmannii" type 1 predominates (80%) and is thought to be acquired from dogs, cats, or pigs. We further examined the zoonotic potential of dogs and cats by amplifying gastric DNA from cats (n = 45) and dogs (n = 10) with primers against "H. heilmannii" ureB and 16S rRNA genes and sequencing the products. Fluorescence in situ hybridization (FISH) with eubacterial and "H. heilmannii"-specific probes was employed to directly visualize "H. heilmannii" types and their intragastric distribution. ureB sequences of "H. heilmannii" amplicons clustered with human and feline isolates of "H. heilmannii" and were distinct from the "H. heilmannii" like organisms (HHLO) H. felis, H. salomonis, and H. bizzozeronii. 16S ribosomal DNA sequences in 20 "H. heilmannii"-infected cats and dogs were distinct from "H. heilmannii" type 1 and "H. suis" and clustered with "H. heilmannii" types 2 and 4. FISH confirmed the presence of "H. heilmannii" types 2 and 4 in dogs but failed to definitively characterize the "H. heilmannii" types present in cats. In infected dogs, "H. heilmannii" inhabited the gastric mucus and glands, and in dogs coinfected with other HHLO it shared the same gastric niche. The results indicate that dogs and cats are predominantly colonized by "H. heilmannii" bacteria that are distinct from type 1 and from "H. suis." As "H. heilmannii" type 1 predominates in people, the zoonotic risk posed by dogs and cats is likely small. PMID- 15131183 TI - DNA microarray format for detection and subtyping of human papillomavirus. AB - A new human papillomavirus (HPV) assay using high-density DNA microarrays is described. An HPV DNA fragment from the 3' end of the E1 gene was amplified and digoxigenin labeled by PCR, and the resulting amplicons were hybridized onto type specific oligonucleotides immobilized on high-density DNA microarrays. For detection, a simple immunohistochemical staining procedure was used with a substrate that has both colorimetric and fluorescent properties. This detection chemistry enables the rapid identification of reactive spots by regular light microscopy and semiquantification by laser scanning. Both single and multiple HPV infections are recognized by this assay, and the corresponding HPV types are easily identified. With this assay, 53 mucosal HPV types were detected and identified. A total of 45 HPV types were identified by a single type-specific probe, whereas the remaining 8 mucosal HPV types could be identified by a specific combination of probes. The simple assay format allows usage of this assay without expensive equipment, making it accessible to all diagnostic laboratories with PCR facilities. PMID- 15131184 TI - Hyperinvasive neonatal group B streptococcus has arisen from a bovine ancestor. AB - The genetic relatedness and evolutionary relationships between group B streptococcus (GBS) isolates from humans and those from bovines were investigated by phylogenetic analysis of multilocus sequence typing data. The collection of isolates consisted of 111 GBS isolates from cows with mastitis and a diverse global collection of GBS isolates from patients with invasive disease (n = 83) and carriers (n = 69). Cluster analysis showed that the majority of the bovine isolates (93%) grouped into one phylogenetic cluster. The human isolates showed greater diversity and clustered separately from the bovine population. However, the homogeneous human sequence type 17 (ST-17) complex, known to be significantly associated with invasive neonatal disease, was the only human lineage found to be clustered within the bovine population and was distinct from all the other human lineages. Split decomposition analysis revealed that the human isolate ST-17 complex, the major hyperinvasive neonatal clone, has recently arisen from a bovine lineage. PMID- 15131185 TI - Correlation between viral loads of cytomegalovirus in blood and bronchoalveolar lavage specimens from lung transplant recipients determined by histology and immunohistochemistry. AB - Cytomegalovirus (CMV) is an important pathogen in lung transplant recipients. Early detection of CMV end-organ disease should help with treatment management. We determined the CMV viral load by hybrid capture in bronchoalveolar lavage (BAL) fluid samples from patients who had undergone lung transplantation. For 39 of these samples (from 25 patients), corresponding transbronchial biopsy samples were available for CMV immunohistochemistry (IHC). The CMV IHC results were interpreted and categorized as positive or negative, and the positive results were subcategorized as typical if cells with both significant nuclear enlargement or Cowdry A-type inclusions and positive staining were present or as atypical if definitive nuclear staining was seen but significant nuclear enlargement was not. Diagnostic CMV viral inclusions were reported in the anatomic diagnosis, based on hematoxylin-eosin staining alone, for three (8%) of the biopsy samples. CMV was detected by IHC in 13 (33%) samples (5 typical, 8 atypical). The median CMV viral load in BAL samples was 0 copies/ml for BAL samples from patients with IHC negative biopsy samples; 47,678 copies/ml for BAL samples from patients with biopsy samples with positive, atypical staining; and 1,548,827 copies/ml for BAL samples from patients with biopsy samples with positive, typical staining (P < 0.001). Compared to routine pathology of biopsy samples, the use of IHC increased the diagnostic yield of CMV. Also, the CMV viral load in BAL fluid samples increased along with immunoreactivity from negative to positive, atypical staining to positive, typical staining. The CMV viral load determined with the end-organ sample, the BAL fluid sample, was higher than the corresponding viral load determined with blood. Both IHC and determination of the CMV viral load in BAL samples may be useful for the detection of individuals at risk for the development of fulminant invasive CMV disease. PMID- 15131186 TI - Use of the DNA flow-thru chip, a three-dimensional biochip, for typing and subtyping of influenza viruses. AB - Influenza A viruses, which are further subtyped on the basis of antigenic differences in external hemagglutinin and neuraminidase glycoproteins, and influenza B viruses are prominent among the viral causes of respiratory diseases and can cause a wide spectrum of illness. Each year these viruses are responsible for recurrent epidemics, frequently in association with genetic variation. There is a requirement for sensitive and rapid diagnostic techniques in order to improve both the diagnosis of infections and the quality of surveillance systems. A new three-dimensional biochip platform (Flow-Thru Chip; MetriGenix) was used to develop a rapid and reliable molecular method for the typing and subtyping of influenza viruses. Oligonucleotide probes immobilized in microchannels of a silicon wafer were selected to recognize multiple fragments of the influenza A virus matrix protein gene; the influenza B virus NS gene; the H1, H3, and H5 hemagglutinin genes; and the N1 and N2 neuraminidase genes. Biotinylated amplicons resulting from either multiplex or random reverse transcription-PCR were hybridized to arrayed oligonucleotides on the influenza virus chip before they were stained with horseradish peroxidase-streptavidin and were imaged by use of a chemiluminescent substrate. The chip analysis procedure, from the time of pipetting of the sample into the chip cartridge to the time of analysis of the results, was performed in less than 5 h. The random PCR exhibited a higher level of performance than the multiplex PCR in terms of the specificity of product hybridization to the influenza virus chip. Analysis of influenza A viruses (H1N1, H3N2, H1N2, and H5N1) and influenza B viruses showed that this microarray-based method is capable of the rapid and unambiguous identification of all types and subtypes of viruses by use of random PCR products. The redundancy of the probes designed for each gene selected yielded an additional criterion of confidence for the subtyping of viruses which are known for antigenic variations in some of their components. PMID- 15131187 TI - Molecular epidemiology of foot-and-mouth disease viruses in the Adamawa province of Cameroon. AB - Foot-and-mouth disease virus (FMDV) causes a highly contagious viral disease of even-toed ungulates and is one of the most important economic diseases of livestock. Most studies of FMDV are done in countries where control measures are being implemented. In contrast, in areas such as sub-Saharan Africa, where FMDV is endemic and new strains are likely to emerge, there are only sporadic submissions to the World Reference Laboratory, Pirbright, United Kingdom. This paper describes the molecular epidemiology of FMDV in the Adamawa province of Cameroon based on a population sample of cattle herds. Serotypes SAT2 and A were isolated in the cross-sectional study. SAT2 isolates were all similar, with phylogenetic distances of <6%, and were most closely related to published sequences of isolates from Eritrea and Saudi Arabia. Serotype A isolates were more variable, with phylogenetic distances of 0 to 11%, and were most closely related to historic isolates from Cameroon. Use of a population-based sample gives a representative sample of virus diversity and will improve our understanding of the evolution of FMDV and its epidemiology. A supplementary study of pigs passing through the railhead collection yard at Ngaoundere detected a serotype O virus. A third pilot longitudinal study monitored viral persistence in three cattle herds over 12 months, and serotype O and A viruses were recovered from a herd 12 months after it was first recorded as being infected with SAT2 virus. The pig type O isolate was not closely related to that recovered from the cattle, suggesting that the pigs had not introduced the O virus into the cattle herds. PMID- 15131188 TI - Systematic 16S rRNA gene sequencing of atypical clinical isolates identified 27 new bacterial species associated with humans. AB - Clinical microorganisms isolated during a 5-year study in our hospital that could not be identified by conventional criteria were studied by 16S rRNA gene sequence analysis. Each isolate yielded a > or =1,400-bp sequence containing <5 ambiguities which was compared with the GenBank 16S rRNA gene library; 1,404 such isolates were tested, and 120 were considered unique (27 isolates) or rare (< or =10 cases reported in the literature) human pathogens. Eleven new species, "Actinobaculum massiliae," "Candidatus Actinobaculum timonae," Paenibacillus sanguinis, "Candidatus Bacteroides massiliae," Chryseobacterium massiliae, "Candidatus Chryseobacterium timonae," Paenibacillus massiliensis, "Candidatus Peptostreptococcus massiliae," "Candidatus Prevotella massiliensis," Rhodobacter massiliensis, and "Candidatus Veillonella atypica" were identified. Sixteen species were obtained from humans for the first time. Our results show the important role that 16S rRNA gene sequence-based bacterial identification currently plays in recognizing unusual and emerging bacterial diseases. PMID- 15131189 TI - Use of beta-lactamase inhibitors in disk tests to detect plasmid-mediated AmpC beta-lactamases. AB - Seeking a simple disk test for detection of organisms producing plasmid-mediated AmpC beta-lactamases, we evaluated the diagnostic utility of the beta-lactamase inhibitors 48-1220 (Ro 48-1220) and LN-2-128. Using NCCLS disk methodology, inhibition zone diameters were determined for five beta-lactam antibiotics tested alone and in combination with 20 microg of either 48-1220 or LN-2-128. Using an increase of > or =4 mm in zone diameter in the presence of an inhibitor as a positive test, cefotetan with LN-2-128 and 48-1220 was adequate for the detection of organisms producing plasmid-mediated AmpCs (specificity of 90% and sensitivity of 100%). PMID- 15131190 TI - Use of the BACTEC MYCO/F LYTIC medium for detection of Brucella melitensis bacteremia. AB - In a prospective volume-controlled study, the sensitivity of the BACTEC MYCO/F LYTIC medium was similar to that of the Peds Plus/F and Plus Aerobic/F vials for detecting Brucella melitensis bacteremia, but the time to detection with the MYCO/F LYTIC medium (101.4 +/- 46.7 h) was significantly longer than that with the combined comparator media (65.5 +/- 18.9 h, P = 0.004). PMID- 15131191 TI - Comparison of VITEK 2 with ITS2-fragment length polymorphism analysis for identification of yeast species. AB - A total of 61 clinical yeast isolates of Candida, Cryptococcus, Blastoschizomyces, and Saccharomyces spp. were used to compare two identification techniques, VITEK 2 and ITS2-fragment length polymorphism analysis (ITS2-FLP), with ID32C as the reference method. ID32C identified 58 isolates correctly. ITS2 FLP with Instagene DNA extraction identified 59 isolates. ITS2-FLP combined with boiling-freezing DNA extraction identified 55 isolates. VITEK 2 identified 41 isolates correctly. PMID- 15131192 TI - Human papillomavirus type 16 viral load is higher in human immunodeficiency virus seropositive women with high-grade squamous intraepithelial lesions than in those with normal cytology smears. AB - Human papillomavirus type 16 (HPV-16) viral load in cervicovaginal lavage samples collected from 66 human immunodeficiency virus-seropositive women was inversely correlated with blood CD4 count (P = 0.002). HPV-16 viral load was 81-fold higher in women with cervical smears suggestive of high-grade lesions (median, 4,425,883 copies/ micro g of DNA) than in women with normal smears (median, 54,576), controlling for age (P = 0.006). PMID- 15131193 TI - Comparison of chlorhexidine and tincture of iodine for skin antisepsis in preparation for blood sample collection. AB - Rates of contamination of blood cultures obtained when skin was prepared with iodine tincture versus chlorhexidine were compared. For iodine tincture, the contamination rate was 2.7%; for chlorhexidine, it was 3.1%. The 0.41% difference is not statistically significant. Chlorhexidine has comparable effectiveness and is safer, cheaper, and preferred by staff, so it is an alternative to iodine tincture. PMID- 15131194 TI - Development and evaluation of a new commercial test allowing the simultaneous detection of noroviruses and sapoviruses by reverse transcription-PCR and microplate hybridization. AB - This work describes the design and initial evaluation of a commercial test allowing the detection of noroviruses and sapoviruses by reverse transcription PCR (RT-PCR) in a single tube followed by microplate hybridization, as well as the detection of PCR inhibitors. The test was shown to be broadly reactive (except for Melksham-like strains), sensitive, and specific and thus should be useful for calicivirus detection in clinical practice. PMID- 15131195 TI - Detection of a rickettsia closely related to Rickettsia aeschlimannii, "Rickettsia heilongjiangensis," Rickettsia sp. strain RpA4, and Ehrlichia muris in ticks collected in Russia and Kazakhstan. AB - Using PCR, we screened 411 ticks from four genera collected in Russia and Kazakhstan for the presence of rickettsiae and ehrlichiae. In Russia, we detected "Rickettsia heilongjiangensis," Rickettsia sp. strain RpA4, and Ehrlichia muris. In Kazakhstan, we detected Rickettsia sp. strain RpA4 and a rickettsia closely related to Rickettsia aeschlimannii. These agents should be considered in a differential diagnosis of tick-borne infections in these areas. PMID- 15131196 TI - Helcococcus ovis isolated from a pulmonary abscess in a horse. AB - Helcococcus ovis, a recently described organism cultured from sheep, was isolated in pure culture from a pulmonary abscess in a horse. This is the first report of this organism in horses and the first report in veterinary medicine to clearly demonstrate a pathogenic role for this organism. PMID- 15131197 TI - Outbreak of Burkholderia cepacia bacteremia in a pediatric hospital due to contamination of lipid emulsion stoppers. AB - We describe a 7-month outbreak of nosocomial Burkholderia cepacia bacteremia involving eight children in a pediatric hospital and the results of epidemiological investigations. A B. cepacia strain genotypically identical to the blood isolates was recovered from the upper surface of capped rubber stoppers of bottles of a commercial lipid emulsion used for parenteral nutrition. PMID- 15131198 TI - Corynebacterium species isolated from bone and joint infections identified by 16S rRNA gene sequence analysis. AB - By the use of 16S rRNA gene sequence analysis we identified 28 of 31 Corynebacterium spp. isolated from bone and joint infections, including species never before isolated in such infections. Phenotypic analysis led to the correct identification of 8 of 31. 16S rRNA gene sequence analysis appears to be a good technique for identification of clinical strains of Corynebacterium spp. PMID- 15131199 TI - Molecular fingerprinting of Mycobacterium bovis subsp. caprae isolates from central Europe. AB - To study the dissemination of Mycobacterium bovis subsp. caprae, 79 European isolates from cattle, humans, and other hosts were examined by spoligotyping and IS6110 restriction fragment length polymorphism (RFLP) analysis. Among a total of 11 different spoligotypes identified, type C1 proved to be predominant (n = 62). Five of the spoligotypes are described for the first time. A total of 43 different RFLP types were identified, thus allowing further differentiation for epidemiological tracking. Isolates from a series of outbreaks in one village proved to be of the same spoligotype and of identical or closely related RFLP types. PMID- 15131200 TI - Further evaluation of a rapid diagnostic test for melioidosis in an area of endemicity. AB - Immunochromatographic test (ICT) kits for the rapid detection of immunoglobulin G (IgG) and IgM antibodies to Burkholderia pseudomallei were compared to the indirect hemagglutination (IHA) assay. In 138 culture-confirmed melioidosis cases, sensitivities were 80, 77, and 88% for IHA, ICT IgG, and ICT IgM, respectively. In a prospective study of 160 consecutive sera samples sent for melioidosis serology, respective specificities were 91, 90, and 69, positive predictive values were 41, 32, and 18, and negative predictive values were 99, 98, and 100%. ICT IgM kits are unreliable for diagnosis of melioidosis, but ICT IgG kits may be useful for diagnosing travelers presenting with possible melioidosis who return from regions where melioidosis is endemic. PMID- 15131201 TI - Rapid detection of point mutations in the gyrA gene of Helicobacter pylori conferring resistance to ciprofloxacin by a fluorescence resonance energy transfer-based real-time PCR approach. AB - As a result of the increasing resistance of Helicobacter pylori against first line antibiotics, other drugs, such as quinolones, will be needed for eradication therapy in the future. We developed a real-time PCR to detect mutations in the gyrA gene associated with ciprofloxacin resistance of H. pylori, thereby contributing to the selection of patients who could be treated by ciprofloxacin based therapy. PMID- 15131202 TI - Comparison of flow cytometric and Alamar Blue tests with the proportional method for testing susceptibility of Mycobacterium tuberculosis to rifampin and isoniazid. AB - The performance of flow cytometry and the microplate Alamar Blue assay in determining susceptibility of Mycobacterium tuberculosis was assessed by testing 150 Brazilian isolates. The overall agreement was 97.3 and 98% for isoniazid and 94.7 and 100% for rifampin by flow cytometry and MABA, respectively. This study was entirely done in a developing country. PMID- 15131203 TI - Collaborative study of the NCCLS and flow cytometry methods for antifungal susceptibility testing of Candida albicans. AB - One hundred clinical isolates of Candida albicans were tested for amphotericin B and fluconazole susceptibilities by the National Committee for Clinical Laboratory Standards (NCCLS) broth microdilution test at center 1 (C1). The same isolates were tested blinded at center 2 (C2) by NCCLS and flow cytometry (FC) methods. The agreement between NCCLS and FC methods ranged from 96 to 99%. PMID- 15131204 TI - Collaborative study of antibiotic medium 3 and flow cytometry for identification of amphotericin B-resistant Candida isolates. AB - Center 1 used the National Committee for Clinical Laboratory Standards M27-A2 method and antibiotic medium 3 (AM3) test to determine amphotericin B resistance in 5 of 30 Candida isolates. These isolates were tested at center 2 by AM3 test and flow cytometry (FC). The agreements (C1-C2) were 90% for AM3 test and FC and 73% for the AM3 tests. PMID- 15131205 TI - Quantitative microbiological study of subgingival plaque by real-time PCR shows correlation between levels of Tannerella forsythensis and Fusobacterium spp. AB - A TaqMan-based real-time PCR assay was established to quantify the periodontopathic bacteria Tannerella forsythensis and Fusobacterium spp. With this assay, the prevalence and proportion of these bacteria in clinical specimens were evaluated. Our preliminary results suggest a positive colocalization of T. forsythensis and Fusobacterium spp. in periodontal pockets. PMID- 15131206 TI - Accuracy and appropriateness of antimicrobial susceptibility test reporting for bacteria isolated from blood cultures. AB - Accurate antimicrobial susceptibility testing (AST) and appropriate reporting of AST results for pathogens isolated from blood cultures are critical functions of the microbiology laboratory. We studied AST performance and reporting from positive blood cultures at hospital microbiology laboratories in Iowa. One hundred sixteen episodes of bacteremia from 14 participating hospitals were examined. We detected AST or identification errors for 18 episodes (16%) and judged reporting of AST results to be inappropriate for 38 episodes (33%). Further study is necessary to determine the impact of testing errors and suboptimal reporting of results on the management of bloodstream infection. PMID- 15131207 TI - Validated 5' nuclease PCR assay for rapid identification of the genus Brucella. AB - A real-time, genus-specific 5' nuclease PCR assay for amplification of a 322-bp fragment of the per gene was developed for rapid (<2 h) identification of Brucella spp. from agar plates. The assay, including an internal amplification control (116 bp), identified Brucella strains (n = 23) and did not detect non Brucella strains (n = 174), indicating its usefulness, particularly for laboratories with stringent quality assurance programs. PMID- 15131208 TI - Presence of bacterial phage-like DNA sequences in commercial Taq DNA polymerase reagents. AB - Many studies have reported the presence of bacterial DNA contamination in commercial Taq DNA polymerase reagents. This is the first report of the presence of phage-like DNA sequences in certain commercial Taq DNA polymerase reagents. Precautions are needed when using amplification reagents with exogenous DNAs. PMID- 15131209 TI - Survey of rodents and ticks in human babesiosis emergence area in Japan: first detection of Babesia microti-like parasites in Ixodes ovatus. AB - Babesia microti-like parasites were detected for the first time in Ixodes ovatus in Hyogo Prefecture, Japan, where two reported types of B. microti-like parasites were recognized in many rodents. Of 80 adult I. ovatus ticks collected, 5 possessed the reported type and 1 possessed a new type of B. microti-like parasite. PMID- 15131210 TI - Asymptomatic and symptomatic excretion of noroviruses during a hospital outbreak of gastroenteritis. AB - During an investigation of a hospital outbreak of norovirus gastroenteritis identified as being caused by a recombinant genogroup II (rGII-3a) strain, fecal specimens collected from asymptomatic staff and patients were tested by nested PCR. A GII-4 norovirus strain, the predominant strain associated with outbreaks in hospitals over the last few years, was detected in 26 and 33% of asymptomatic staff and patients, respectively. No rGII-3a (Harrow/Mexico) norovirus strains were detected in the samples of asymptomatic staff or patients. PMID- 15131211 TI - Detection of Candida albicans mRNA in archival histopathology samples by reverse transcription-PCR. AB - The feasibility of detecting Candida albicans mRNA in formalin-fixed paraffin embedded archival human histopathology specimens by reverse transcription-PCR (RT PCR) was investigated. RT with gene-specific primers was used to detect five single-copy C. albicans gene transcripts, including those of two housekeeping genes, in oral candidiasis samples up to 8 years of age. PMID- 15131212 TI - Correlation between Helicobacter pylori OipA protein expression and oipA gene switch status. AB - Polyclonal antisera to either a synthetic OipA peptide or a recombinant OipA protein detected OipA expression in Helicobacter pylori and correlated with functional oipA status determined by PCR sequence (sensitivity and specificity of >94%). Immunoblotting is a simple and accurate method for detecting expression of the important virulence factor OipA. PMID- 15131213 TI - Use of the Vitek-1 and Vitek-2 systems for detection of constitutive and inducible macrolide resistance in group B streptococci. AB - A prospective study of erythromycin and clindamycin resistance was performed with 304 consecutive group B streptococci (GBS) isolates. According to two automated susceptibility testing systems, Vitek-1 and Vitek-2, and double-disk agar diffusion, 79.9% were susceptible to both erythromycin and clindamycin. However, for macrolide-lincosamide-streptogramin B-inducible isolates, the accuracies of the Vitek-1 and Vitek-2 systems were 5.6 and 94.4%. In light of these results, we recommend that GBS be routinely tested using the double-disk diffusion method. PMID- 15131214 TI - Molecular evidence of Anaplasma phagocytophilum in Ixodes ricinus ticks and wild animals in Austria. AB - Real-time PCR analysis of a groESL heat shock operon segment showed the presence of two genetic lineages of Anaplasma phagocytophilum in Ixodes ricinus ticks as well as one variant in wild red and roe deer, the latter supposedly representing the natural reservoir of one variant of A. phagocytophilum. PMID- 15131215 TI - AFMP2 encodes a novel immunogenic protein of the antigenic mannoprotein superfamily in Aspergillus fumigatus. AB - We cloned the Aspergillus fumigatus mannoprotein 2 (AFMP2) gene, which encodes a novel immunogenic protein (Afmp2p) of the antigenic mannoprotein superfamily, in A. fumigatus. Sequence analysis revealed that Afmp2p has 510 amino acid residues, with a predicted molecular mass of 51.5 kDa. Afmp2p has a putative N-terminal signal peptide, a putative C-terminal glycosylphosphatidylinositol membrane attachment signal sequence, and an upstream GAA cleavage site commonly used for cytoplasmic membrane attachment and implicated in fungal cell wall assembly. Upstream of the GAA cleavage site, Afmp2p contains a 302-amino-acid serine- and threonine-rich region as a site for potential O-glycosylation. Within this serine and threonine-rich region, 13 repeats of ETSTPCE(T)(n) were observed. Western blot analysis of Afmp2p in A. fumigatus fungal cell lysate and culture supernatant and immunogold staining and electron microscopy showed that Afmp2p is predominantly secreted into the culture supernatant, whereas only minimal amounts can be detected in the cell lysate and cell wall. Finally, it was observed that patients with aspergilloma and invasive aspergillosis due to A. fumigatus develop a specific antibody response against recombinant Afmp2p. The abundance of Afmp2p in secreted form, its minimal cross-reactivity with Afmp1p, and the presence of an antibody response against Afmp2p in patients with A. fumigatus infections suggest that Afmp2p is a good candidate for complementing Afmp1p in serodiagnosis of A. fumigatus infections. PMID- 15131216 TI - Use of a panfungal PCR assay for detection of fungal pathogens in a commercial blood culture system. AB - A panfungal PCR assay was used to evaluate the ability of the ESP blood culture system to detect fungemia. The results showed that the ESP system is reliable for the detection of fungi and showed the applicability of using a molecular-based assay as a potential rapid and reliable method for the identification of fungi. PMID- 15131217 TI - Culture-independent species typing of neotropical Leishmania for clinical validation of a PCR-based assay targeting heat shock protein 70 genes. AB - PCR-restriction fragment length polymorphism analysis of heat shock protein 70 genes discriminates most neotropical Leishmania species, as well as Trypanosoma cruzi. The assay, combined with capillary electrophoresis in a microchip device, may be applied directly on clinical samples with a high sensitivity, hence supporting clinical and epidemiological monitoring of leishmaniasis. PMID- 15131218 TI - Frequent detection of viral nucleic acids in heart valve tissue. AB - Due to a paucity of published data concerning the prevalence of viral nucleic acid in homografts, we analyzed tissue from 30 donor hearts for the presence of viral genome sequences of enteroviruses, adenoviruses, human cytomegalovirus, and influenza virus using different PCR techniques. Viral DNA was amplified in 64 and 52% of the subvalvular myocardial tissue and non-coronary valve samples, respectively. These findings, compared with clinical history and histologic and serologic analysis, demonstrate the importance of viral safety measures in heart valve banking. PMID- 15131219 TI - Use of the omp50 gene for identification of Campylobacter species by PCR. AB - We studied the prevalence of the omp50 gene and the Omp50 protein in Campylobacter strains. Immunodetection assays and DNA-DNA hybridizations showed that most C. coli strains tested were negative and most C. jejuni and C. lari strains tested were positive. A PCR assay was developed, using the omp50 gene as a species-specific target. We propose a combination of a hippurate test and an omp50 assay to perform identification of Campylobacter species. PMID- 15131220 TI - Detection of specific antibodies to severe acute respiratory syndrome (SARS) coronavirus nucleocapsid protein for serodiagnosis of SARS coronavirus pneumonia. AB - We report the evaluation of recombinant severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) nucleocapsid protein enzyme-linked immunosorbent assay (ELISA)-based antibody tests for serodiagnosis of SARS-CoV pneumonia and compare the sensitivities and specificities of this ELISA for detection of immunoglobulin G (IgG), IgM, IgA, and their combinations with serum samples from 149 healthy blood donors who donated blood 3 years ago as controls and 106 SARS-CoV pneumonia patients in Hong Kong. The specificities of the ELISA for IgG, IgM, and IgA detection were 95.3, 96.6, and 96.6%, respectively, with corresponding sensitivities of 94.3, 59.4, and 60.4%, respectively. The present ELISA appears to be a sensitive test for serodiagnosis of SARS-CoV pneumonia, is much more economical and less labor-intensive than the indirect immunofluorescence assay, and does not require cultivation of SARS-CoV. PMID- 15131221 TI - First serologic evidence of human spotted fever group rickettsiosis in Korea. AB - To investigate the prevalence of spotted fever group rickettsioses in Korea, a serosurvey of Japanese spotted fever rickettsiosis in patients with acute febrile illness was conducted with an indirect immunofluorescence assay. Overall, 19.88% of the patients were found to have polyvalent antibody against Rickettsia japonica. This study is the first documentation of spotted fever group rickettsiosis in Korea. PMID- 15131222 TI - Characterization of Salmonella enterica serotype Typhimurium isolates from human, food, and animal sources in the Republic of Ireland. AB - A potential epidemic clone of Salmonella enterica serotype Typhimurium DT104, and the possible emergence of S. enterica serotype Typhimurium DT104b, has been identified from the characterization of 67 S. enterica serotype Typhimurium strains from three sources, human gastroenteritis isolates, isolates from food samples, and veterinary isolates, by antimicrobial resistance profiling, phage typing, and pulsed-field gel electrophoresis. Resistance to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline was found in 77.6% of these strains. PMID- 15131223 TI - Molecular epidemiology of the fsr locus and of gelatinase production among different subsets of Enterococcus faecalis isolates. AB - We examined 215 Enterococcus faecalis isolates and found that neither the two component regulatory locus fsr (E. faecalis regulator) nor gelatinase production was more common in disease-associated isolates than in isolates colonizing healthy individuals (ca. 60 to 65%). The majority of gelatinase-negative isolates, including 14 endocarditis isolates (of 80 isolates tested), contained the previously described 23.9-kb deletion and lacked fsrA and fsrB. While these findings indicate that neither fsr nor gelatinase is required for E. faecalis to cause infection, this study did not address whether fsr or gelatinase affects the severity of disease, as it does in animal models. PMID- 15131224 TI - Meta-analysis of BACTEC MGIT 960 and BACTEC 460 TB, with or without solid media, for detection of mycobacteria. AB - In a meta-analysis of 10 studies, the BACTEC 960/MGIT and BACTEC 460 systems showed a sensitivity and specificity in detecting mycobacteria (1,381 strains from 14,745 clinical specimens) of 81.5 and 99.6% and 85.8 and 99.9%, respectively. Combined with solid media, the sensitivity of the two systems increased to 87.7 and 89.7%, respectively. PMID- 15131225 TI - Glycerophosphodiester phosphodiesterase gene (glpQ) of Borrelia lonestari identified as a target for differentiating Borrelia species associated with hard ticks (Acari:Ixodidae). AB - A glpQ ortholog was identified in DNA from Borrelia lonestari-positive Amblyomma americanum, providing further evidence that B. lonestari is more closely related to the relapsing fever group spirochetes than to borreliae that cause Lyme disease. This finding provides a basis for developing diagnostic assays to differentiate species of borrelia transmitted by hard ticks. PMID- 15131226 TI - Phylogenetic relationships among virulent Newcastle disease virus isolates from the 2002-2003 outbreak in California and other recent outbreaks in North America. AB - Isolates from the 2002-2003 virulent Newcastle disease virus (v-NDV) outbreak in southern California, Nevada, Arizona, and Texas in the United States were compared to each other along with recent v-NDV isolates from Mexico and Central America and reference avian paramyxovirus type 1 strains. Nucleotide sequencing and phylogenetic analyses were conducted on a 1,195-base genomic segment composing the 3' region of the matrix (M) protein gene and a 5' portion of the fusion (F) protein gene including the M-F intergenic region. This encompasses coding sequences for the nuclear localization signal of the M protein and the F protein cleavage activation site. A dibasic amino acid motif was present at the predicted F protein cleavage activation site in all v-NDVs, including the California 2002-2003, Arizona, Nevada, Texas, Mexico, and Central America isolates. Phylogenetic analyses demonstrated that the California 2002-2003, Arizona, Nevada, and Texas viruses were most closely related to isolates from Mexico and Central America. An isolate from Texas obtained during 2003 appeared to represent a separate introduction of v-NDV into the United States, as this virus was even more closely related to the Mexico 2000 isolates than the California, Arizona, and Nevada viruses. The close phylogenetic relationship between the recent 2002-2003 U.S. v-NDV isolates and those viruses from countries geographically close to the United States warrants continued surveillance of commercial and noncommercial poultry for early detection of highly virulent NDV. PMID- 15131227 TI - Chronic cholangitis caused by Bordetella hinzii in a liver transplant recipient. AB - Bordetella hinzii was isolated in four biliary specimens collected over 6 months from a liver transplant recipient with cholangitis. The isolates were resistant to most beta-lactam antibiotics and fluoroquinolones. Molecular typing was performed by pulsed-field gel electrophoresis. These data add cholangitis to the spectrum of disease manifestations caused by B. hinzii. PMID- 15131228 TI - Brain abscess due to Gemella haemolysans. AB - We present a case of brain abscess due to Gemella haemolysans and Bacteroides species in a 60-year-old-immunocompetent man who underwent dental procedures. The patient completely recovered following intravenous therapy with ampicillin and metronidazole for 6 weeks. PMID- 15131229 TI - Clinical case of endocarditis due to Trichosporon inkin and antifungal susceptibility profile of the organism. AB - A fatal case of Trichosporon inkin prosthetic endocarditis is reported. The isolation sites and susceptibility profiles of 10 other isolates are also reviewed. Four strains were recovered from cutaneous or subcutaneous samples, four were recovered from urine, one was recovered from peritoneal liquid, and one was recovered from bone. Voriconazole and amphotericin B had the most potent activities in vitro against the isolates, with MIC geometric means of 0.11 and 0.30 microg/ml, respectively. PMID- 15131230 TI - Invasive pneumococcal infection in a healthy infant caused by two different serotypes. AB - We present a case of invasive pneumococcal infection in a healthy 10-month-old infant from whom Streptococcus pneumoniae serotype 23F was isolated from the blood and serotype 23B was isolated from the cerebrospinal fluid. Both serotypes were penicillin nonsusceptible. Pulsed-field gel electrophoresis analysis demonstrated that the two serotypes had distinct DNA patterns, indicating that infection did not occur as a result of capsular transformation but as a result of a mixed infection with two distinct pneumococcal serotypes. PMID- 15131231 TI - Posttransplantation disseminated coccidioidomycosis acquired from donor lungs. AB - A North Carolinian developed fatal coccidioidomycosis immediately after bilateral lung transplantation. The donor had previously traveled to Mexico, and the recipient had no travel history to an area where Coccidioides immitis is endemic. Immunosuppressive therapy of the transplant recipient likely reactivated latent Coccidioides infection in the donor lungs, leading to posttransplant coccidioidomycosis. PMID- 15131232 TI - Identification of 2,600 clinical methicillin-resistant Staphylococcus aureus strains in The Netherlands yielded sporadic cases of strains negative for the species-specific Sa442 gene fragment. PMID- 15131233 TI - Antigenic cross-reactivity between the nucleocapsid protein of severe acute respiratory syndrome (SARS) coronavirus and polyclonal antisera of antigenic group I animal coronaviruses: implication for SARS diagnosis. PMID- 15131234 TI - Weak agreement between Antivirogram and Phenosense assays in predicting reduced susceptibility to antiretroviral drugs. PMID- 15131235 TI - Investigating infectious keratitis. PMID- 15131236 TI - Virological interpretations of dengue disease spectrum in infants in Chennai, Tamil Nadu, India, need reevaluation. PMID- 15131237 TI - Revision of the Starling principle: new views of tissue fluid balance. PMID- 15131238 TI - Short-term bed rest impairs amino acid-induced protein anabolism in humans. AB - Diminished muscular activity is associated with alterations of protein metabolism. The aim of this study was to evaluate the effect of short-term muscle inactivity on regulation of whole-body protein deposition during amino acid infusion to simulate an experimental postprandial state. We studied nine healthy young volunteers at the end of 14 day periods of strict bed rest and of controlled ambulation using a cross-over design. Subjects received a weight maintaining diet containing 1 g protein kg(-1) day(-1). l[1-(13)C]leucine was used as a marker of whole-body protein kinetics in the postabsorptive state and during a 3 h infusion of an amino acid mixture (0.13 g amino acid (kg lean body mass)(-1) h(-1)). In the postabsorptive state, bed rest decreased (P < 0.05) the rate of leucine disposal (R(d)) to protein synthesis and tended to decrease leucine rate of appearance (R(a)) from proteolysis, whereas the rate of leucine oxidation did not change significantly. Amino acid infusion increased leucine R(d) to protein synthesis and oxidation and decreased leucine R(a) from proteolysis in both the bed rest and ambulatory conditions. Changes from basal in leucine R(d) to protein synthesis were lower (P < 0.05) during bed rest than those in the ambulatory period, whereas changes in leucine R(a) from proteolysis and oxidation were not significantly different. During amino acid infusion, net leucine deposition into body protein was 8 +/- 3% lower during bed rest than during the ambulatory phase. In conclusion, short-term bed rest leads to reduced stimulation of whole-body protein synthesis by amino acid administration. Results of this study were, in part, presented at the meeting, Experimental Biology, 2004, Washington DC. PMID- 15131239 TI - Development affects in vitro vascular tone and calcium sensitivity in ovine cerebral arteries. AB - We have shown recently that development from neonatal to adult life affects cerebrovascular tone of mouse cerebral arteries through endothelium-derived vasodilatory mechanisms. The current study tested the hypothesis that development from fetal to adult life affects cerebral artery vascular smooth muscle (VSM) [Ca(2+)](i) sensitivity and tone through a mechanism partially dependent upon endothelium-dependent signalling. In pressurized resistance sized cerebral arteries ( approximately 150 microm) from preterm (95 +/- 2 days gestation (95 d)) and near-term (140 +/- 2 days gestation (140 d)) fetuses, and non-pregnant adults, we measured vascular diameter (microm) and [Ca(2+)](i) (nm) as a function of intravascular pressure. We repeated these studies in the presence of inhibition of nitric oxide synthase (NOS; with l-NAME), cyclo-oxygenase (COX; with indomethacin) and endothelium removal (E-). Cerebrovasculature tone (E+) was greater in arteries from 95 d fetuses and adults compared to 140 d sheep. Ca(2+) sensitivity was similar in 95 d fetuses and adults, but much lower in 140 d fetuses. Removal of endothelium resulted in a reduction in lumen diameter as a function of pressure (greater tone) in all treatment groups. [Ca(2+)](i) sensitivity differences among groups were magnified after E-. NOS inhibition decreased diameter as a function of pressure in each age group, with a significant increase in [Ca(2+)](i) to pressure ratio only in the 140 d fetuses. Indomethacin increased tone and increased [Ca(2+)](i) in the 140 d fetuses, but not the other age groups. Development from near-term to adulthood uncovered an interaction between NOS- and COX-sensitive substances that functioned to modulate artery diameter but not [Ca(2+)](i). This study suggests that development is associated with significant alterations in cerebral vascular smooth muscle (VSM), endothelium, NOS and COX responses to intravascular pressure. We speculate that these changes have important implications in the regulation of cerebral blood flow in the developing organism. PMID- 15131241 TI - Differential involvement of multidrug resistance-associated protein 1 and P glycoprotein in tissue distribution and excretion of grepafloxacin in mice. AB - The involvement of multidrug resistance-associated protein 1 (Mrp1) and P glycoprotein (mdr1) in the tissue distribution and excretion of grepafloxacin (GPFX), a fluoroquinolone antibiotic, was investigated using gene-deficient mice [mdr1a(-/-), mdr1a/1b(-/-), and mrp1(-/-)]. The plasma concentration-time profile of GPFX in mrp1(-/-) was nearly identical to that in mrp1(+/+), whereas that in mdr1a/1b(-/-) was higher than that in mdr1a/1b(+/+). The urinary clearance of GPFX in mdr1a/1b(-/-) was lower than that in mdr1a/1b(+/+), suggesting that the urinary excretion of GPFX is at least partially mediated by mdr1. The tissue-to plasma concentration ratios during the beta-phase (K(p beta),) was significantly higher in the heart, trachea, kidney, spleen, and brown fat of mrp1(-/-) than those in mrp1(+/+). In MRP1-transfected LLC-PK1 cells, the efflux of GPFX after preloading into the cells was higher than that observed in the parent cell lines. These results suggest that GPFX is a substrate of MRP1 and that its distribution to these tissues might be limited by Mrp1. On the other hand, a higher K(p beta), and of GPFX in mdr1a(-/-) mdr1a/1b(-/-) compared with mdr1a/1b(+/+) was observed only in the brain. GPFX was efficiently distributed to the lung parenchyma cells and pulmonary airspaces, including the epithelial lining fluid and macrophages that are the pharmacological target of GPFX, although the contribution of Mdr1 and Mrp1 to such distribution seems to be minor. Thus, the present findings reveal that the disposition of GPFX is at least in part governed by these two ABC transporters and that both Mrp1 and Mdr1 are involved in the limited distribution of GPFX to the distinct tissues, including pharmacological and/or toxicological targets by an active efflux mechanism. PMID- 15131242 TI - Nonsteroidal anti-inflammatory drugs potentiate 1-methyl-4-phenylpyridinium (MPP+)-induced cell death by promoting the intracellular accumulation of MPP+ in PC12 cells. AB - In this study, we investigated the effects of nonsteroidal anti-inflammatory drugs (NSAIDs) on 1-methyl-4-phenylpyridinium (MPP(+))-induced cell death in PC12 cells. Coincubation of PC12 cells with indomethacin, ibuprofen, ketoprofen, or diclofenac, but not aspirin or N-[2-(cyclohexyloxy)-4 nitrophenyl]methanosulfonamide (NS-398), significantly potentiated the MPP(+) induced cell death. In contrast, these NSAIDs had no effect on rotenone-induced cell death. The potentiating actions of these NSAIDs were not suppressed by treatment with phenyl-N-butyl-nitrone, a radical scavenger; N-acetyl-l-cysteine, an antioxidant; Ac-DEVD-CHO, a selective caspase-3 inhibitor; or 2-chloro-5-nitro N-phenylbenzamide (GW9662), a selective antagonist of peroxisome proliferator activated receptor gamma. Furthermore, we observed that DNA fragmentation, which is one of the hallmarks of apoptosis, was not induced by coincubation with MPP(+) and NSAIDs. We confirmed that coincubation of PC12 cells with 30 microM MPP(+) and 100 microM indomethacin, ibuprofen, ketoprofen, or diclofenac led to a significant increase in the accumulation of intracellular MPP(+) compared with incubation with 30 microM MPP(+) alone. In addition, these NSAIDs markedly reduced the efflux of MPP(+) from PC12 cells. (3-(3-(2-(7-Chloro-2-quinolinyl) ethenyl) phenyl ((3-dimethyl amino-3oxo-propyl) thio) methyl) propanoic acid (MK 571), which is an inhibitor of multidrug resistance proteins (MRPs), mimicked the NSAIDs-induced effects, increasing cell toxicity and promoting the accumulation of MPP(+). Moreover, some types of MRPs' mRNA were detected in PC12 cells. These results suggest that some NSAIDs might cause a significant increase in the intracellular accumulation of MPP(+) via the suppression of reverse transport by the blockade of MRP, resulting in the potentiation of MPP(+)-induced cell death. PMID- 15131240 TI - Lactate metabolism: a new paradigm for the third millennium. AB - For much of the 20th century, lactate was largely considered a dead-end waste product of glycolysis due to hypoxia, the primary cause of the O2 debt following exercise, a major cause of muscle fatigue, and a key factor in acidosis-induced tissue damage. Since the 1970s, a 'lactate revolution' has occurred. At present, we are in the midst of a lactate shuttle era; the lactate paradigm has shifted. It now appears that increased lactate production and concentration as a result of anoxia or dysoxia are often the exception rather than the rule. Lactic acidosis is being re-evaluated as a factor in muscle fatigue. Lactate is an important intermediate in the process of wound repair and regeneration. The origin of elevated [lactate] in injury and sepsis is being re-investigated. There is essentially unanimous experimental support for a cell-to-cell lactate shuttle, along with mounting evidence for astrocyte-neuron, lactate-alanine, peroxisomal and spermatogenic lactate shuttles. The bulk of the evidence suggests that lactate is an important intermediary in numerous metabolic processes, a particularly mobile fuel for aerobic metabolism, and perhaps a mediator of redox state among various compartments both within and between cells. Lactate can no longer be considered the usual suspect for metabolic 'crimes', but is instead a central player in cellular, regional and whole body metabolism. Overall, the cell to-cell lactate shuttle has expanded far beyond its initial conception as an explanation for lactate metabolism during muscle contractions and exercise to now subsume all of the other shuttles as a grand description of the role(s) of lactate in numerous metabolic processes and pathways. PMID- 15131243 TI - The adenosine transporter, mENT1, is a target for adenosine receptor signaling and protein kinase Cepsilon in hypoxic and pharmacological preconditioning in the mouse cardiomyocyte cell line, HL-1. AB - Brief exposure of the heart to hypoxia results in less cellular damage after subsequent hypoxia, an effect known as preconditioning (PC). PC has been widely studied but is still not fully understood. Adenosine (Ado), adenosine receptors, and protein kinase C (PKC) have been implicated as integral components of PC. Adenosine (nucleoside) transporters (NTs) facilitate flux of Ado across cell membranes, but their role in PC is unknown. Therefore, we used the murine cardiomyocyte cell line, HL-1, and asked if there was feedback regulation of NTs by Ado, Ado receptors, and PKC following either hypoxic or pharmacological PC. Activation (by specific agonists) of A1 or A3 Ado receptors or PKC resulted in PC in HL-1. The A1 (but not A3) receptor is coupled to PKCepsilon, and activation of PKCepsilon (by specific peptide agonist) resulted in PC. Moreover, PKCepsilon stimulates Ado uptake via the predominant NT in HL-1, mouse equilibrative nucleoside transporter 1 (mENT1). Studies in primary neonatal mouse cardiomyocytes confirmed our observations in HL-1 cells. Hypoxic challenge led to a rapid increase in, and efflux of, intracellular Ado from cells, which was blocked by NT inhibitors (dipyridamole/nitrobenzylthioinosine). Moreover, NT inhibition during hypoxia or PC was highly protective, suggesting that Ado loss contributes to decreased cell viability. Our data suggest that hypoxic challenge causes an efflux of Ado via ENTs, activation of A1 and/or A3 receptors, signaling through PKCepsilon, and activation of ENT1. Since Ado is required for ATP synthesis on reperfusion, this feedback regulation of mENT1 would promote reuptake of Ado. PMID- 15131244 TI - Strong protein adduct trapping accompanies abolition of acrolein-mediated hepatotoxicity by hydralazine in mice. AB - Acrolein is a highly reactive alpha,beta-unsaturated aldehyde that readily alkylates nucleophilic centers in cell macromolecules. Typically, such reactions proceed via Michael addition chemistry, forming adducts that retain an electrophilic carbonyl group. Since these species participate in secondary deleterious reactions, we hypothesize that inactivation of carbonyl adducts may attenuate acrolein toxicity. Indeed, we recently established that the nucleophilic antihypertensive drug hydralazine readily "traps" acrolein adducts in cell proteins and strongly suppresses acrolein-mediated toxicity in isolated hepatocytes. This work sought to determine whether hydralazine prevents the in vivo hepatotoxicity of the acrolein precursor allyl alcohol in whole mice and whether adduct trapping accompanies any such hepatoprotection. Mice received allyl alcohol alone or in conjunction with several doses of hydralazine. Four hours later, mice were sacrificed to allow for the determination of liver enzymes in plasma as markers of hepatic injury, whereas livers were assessed for glutathione and hydralazine-stabilized protein adducts. Hydralazine afforded strong, dose-dependent protection against the increases in plasma marker enzymes but not the hepatic glutathione depletion produced by allyl alcohol. Western blotting revealed intense, dose-dependent adduct trapping by hydralazine in numerous liver proteins over a broad 26- to 200-kDA mass range. In keeping with these findings, immunohistochemical analysis of liver slices indicated diffuse, extranuclear adduct trapping by hydralazine that was uniformly distributed across the liver lobule, with partial localization in parenchymal cell membranes. These findings concur with our hypothesis that hydralazine readily inactivates reactive carbonyl-retaining protein adducts formed by acrolein, thereby preventing secondary reactions that trigger cellular death. PMID- 15131245 TI - SR147778 [5-(4-bromophenyl)-1-(2,4-dichlorophenyl)-4-ethyl-N-(1-piperidinyl)-1H pyrazole-3-carboxamide], a new potent and selective antagonist of the CB1 cannabinoid receptor: biochemical and pharmacological characterization. AB - Based on binding, functional, and pharmacological data, this study introduces SR147778 [5-(4-bromophenyl)-1-(2,4-dichloro-phenyl)-4-ethyl-N-(1-piperidinyl)-1H pyrazole-3-carboxamide] as a highly potent, selective, and orally active antagonist for the CB1 receptor. This compound displays nanomolar affinity (Ki = 0.56 and 3.5 nM) for both the rat brain and human CB1 recombinant receptors, respectively. It has low affinity (Ki = 400 nM) for both the rat spleen and human CB2 receptors. Furthermore, it shows no affinity for any of the over 100 targets investigated (IC50 > 1 microM). In vitro, SR147778 antagonizes the inhibitory effects of CP 55,940 [(1R,3R,4R)-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-4-(3 hydroxypropyl)cyclohexan-1-ol] on both the mouse vas deferens contractions (pA2 value = 8.1) and on forskolin-stimulated adenylyl cyclase activity in the U373 MG cell lines (pA2 value = 8.2) but not in Chinese hamster ovary (CHO) cells permanently expressing the human peripheral cannabinoid receptor (hCB2). SR147778 is able to block the mitogen-activated protein kinase activity induced by CP 55,940 in the CHO cell line expressing human brain cannabinoid receptor (IC50 = 9.6 nM) but was inactive in cells expressing hCB2. After oral administration, SR147778 displaced the ex vivo [3H]-CP 55,940 binding to mouse brain membranes (ED50 = 3.8 mg/kg) with a long duration of action, whereas it did not interact with the CB2 receptor expressed in the mouse spleen. Using different routes of administration, SR147778 (0.3-3 mg/kg) is shown to antagonize pharmacological effects (hypothermia, analgesia, and gastrointestinal transit) induced by R-(+) (2,3-dihydro-5-methyl-3-[[4-morpholinyl]methyl] pyrol [1,2,3-de]-1,4-benzoxazin-6 yl)(1-naphthalenyl) methanone in mice. Finally, per se, SR147778 (0.3-10 mg/kg) is able to reduce ethanol or sucrose consumption in mice and rats and food intake in fasted and nondeprived rats. PMID- 15131246 TI - Contribution of serotonin (5-hydroxytryptamine; 5-HT) 5-HT2 receptor subtypes to the hyperlocomotor effects of cocaine: acute and chronic pharmacological analyses. AB - The role of serotonin (5-hydroxytryptamine; 5-HT) 5-HT2 receptor subtypes (5 HT2AR, 5-HT2BR, and 5-HT2CR) in acute cocaine-evoked hyperactivity was compared with their contribution to the development and expression of locomotor sensitization upon repeated, intermittent treatment with cocaine (10 mg/kg/day for 5 days) in male Wistar rats. Cocaine-evoked hyperactivity was significantly enhanced by pretreatment with the preferential 5-HT2AR agonist 1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane (DOI) and the 5-HT2CR antagonist SDZ SER-082 [(+)-cis 4,5,7a,8,9,10,11,11a-octahydro-7H-10-methylindolo(1,7-BC)(2,6) naphthyridine fumarate]. The 5-HT2AR antagonist SR 46349B [1(Z)-[2-(dimethylamino)ethoxyimino] 1(2-fluorophenyl)-3-(4-hydroxyphenyl)-2(E)-propene] and the preferential 5-HT2CR agonist MK 212 [6-chloro-2-(1-piperazinyl)pyrazine HCl] (2 mg/kg) significantly attenuated acute cocaine-evoked hyperactivity; however, a lower dose of MK 212 (0.3 mg/kg) enhanced cocaine-evoked hyperactivity. The 5-HT2BR agonist BW 723C86 (1-[5-(2-thienylmethoxy)-1H-3-indolyl]propan-2-amine HCl) and the 5-HT2BR antagonist SB 204741 [N-(1-methyl-5-indolyl)-N'-(3-methyl-5-isothiazolyl) urea] had no effect on cocaine-evoked hyperactivity. Repeated treatment with cocaine alone resulted in a 2-fold increase in hyperactivity upon challenge with cocaine 5 days after termination of the cocaine regimen (sensitization). The 5-HT2AR antagonist SR 46349B also blocked cocaine-evoked hyperactivity following repeated cocaine treatment, whereas the other 5-HT2R ligands were ineffective. When any of the 5-HT2R ligands was coadministered with cocaine during the treatment regimen (10 mg/kg/day for 5 days), the development of sensitization was unchanged as measured by the level of cocaine-evoked hyperactivity upon challenge 5 days after termination of the treatment. The present study implies that 5-HT2AR and 5-HT2CR exert oppositional influence upon hyperactivity evoked by acute administration of cocaine; this balance is altered following repeated cocaine administration. PMID- 15131247 TI - Regulation of ovule development. PMID- 15131248 TI - Interactions of mitochondrial and nuclear genes that affect male gametophyte development. PMID- 15131249 TI - Stamen structure and function. PMID- 15131250 TI - Understanding apomixis: recent advances and remaining conundrums. PMID- 15131251 TI - The genetic, developmental, and molecular bases of fruit size and shape variation in tomato. PMID- 15131252 TI - Tomato heat stress transcription factor HsfB1 represents a novel type of general transcription coactivator with a histone-like motif interacting with the plant CREB binding protein ortholog HAC1. AB - In contrast with the class A heat stress transcription factors (HSFs) of plants, a considerable number of HSFs assigned to classes B and C have no evident function as transcription activators on their own. However, in the following article, we provide evidence that tomato (Lycopersicon peruvianum) HsfB1 represents a novel type of coactivator cooperating with class A HSFs (e.g., with tomato HsfA1). Provided the appropriate promoter architecture, the two HSFs assemble into an enhanceosome-like complex, resulting in strong synergistic activation of reporter gene expression. Moreover, HsfB1 also cooperates in a similar manner with other activators, for example, with the ASF1/2 enhancer binding proteins of the 35S promoter of Cauliflower mosaic virus or with yet unidentified activators controlling housekeeping gene expression. By these effects, HsfB1 may help to maintain and/or restore expression of certain viral or housekeeping genes during ongoing heat stress. The coactivator function of HsfB1 depends on a histone-like motif in its C-terminal domain with an indispensable Lys residue in the center (GRGKMMK). This motif is required for recruitment of the plant CREB binding protein (CBP) ortholog HAC1. HsfA1, HsfB1, and HAC1/CBP form ternary complexes in vitro and in vivo with markedly enhanced efficiency in promoter recognition and transcription activation in plant and mammalian (COS7) cells. Using small interfering RNA-mediated knock down of HAC1 expression in Arabidopsis thaliana mesophyll protoplasts, the crucial role for the coactivator function of HsfB1 was confirmed. PMID- 15131253 TI - Effect of lesions on the dynamics of DNA on the picosecond and nanosecond timescales using a polarity sensitive probe. AB - This paper explores the effects of structural modifications on the fast dynamics of DNA and the ability of time-resolved Stokes shift spectroscopy to measure those changes. The time-resolved Stokes shift of a synthetic coumarin base-pair replacement within an oligomer is measured between 40 ps and 40 ns. Comparisons are made between 17mers without modification, with a deleted base near the coumarin and with the coumarin placed near the end of the oligomer. The deletion of a next-to-nearest-neighbor base pair does not change the subnanosecond dynamics, but does cause an additional motion with a time constant of approximately 20 ns. A candidate for this motion is the flipping of the abasic sugar out of the helix and the concomitant intrusion of water into the interior of the helix. A nearby chain end causes little change in the dynamics after 1 ns but leads to a reduction in the amplitude of the dynamics between 40 ps and 1 ns. We suggest that at the chain end, where DNA on one side of the probe has been replaced by water, the charge- stabilizing dynamics have the same overall amplitude, but that much of the relaxation occurs before the start of the measurement time window. PMID- 15131254 TI - Targeted transcriptional repression of Gfi1 by GFI1 and GFI1B in lymphoid cells. AB - Growth factor independence-1 (GFI1) and GFI1B are closely related, yet differentially expressed transcriptional repressors with nearly identical DNA binding domains. GFI1 is upregulated in the earliest thymocyte precursors, while GFI1B expression is restricted to T lymphopoiesis stages coincident with activation. Transgenic expression of GFI1 potentiates T-cell activation, while forced GFI1B expression decreases activation. Both mice and humans with mutant Gfi1 display lymphoid abnormalities. Here we describe autoregulation of Gfi1 in primary mouse thymocytes and a human T-cell line. GFI1 binding to cis-element sequences conserved between rat, mouse and human Gfi1 mediates direct and potent transcriptional repression. In addition, dramatic regulation of Gfi1 can also be mediated by GFI1B. These data provide the first example of a gene directly targeted by GFI1 and GFI1B. Moreover, they support a role for auto- and trans regulation of Gfi1 by GFI1 and GFI1B in maintaining the normal expression patterns of Gfi1, and suggest that GFI1B may indirectly affect T-cell activation through repression of Gfi1. PMID- 15131255 TI - The Fen1 extrahelical 3'-flap pocket is conserved from archaea to human and regulates DNA substrate specificity. AB - Fen1 is a key enzyme for the maintenance of genetic stability in archaea and eukaryotes and is classified as a tumor suppressor. Very recent structural data obtained from Archaeoglobus fulgidus Fen1 suggest that an extrahelical 3'-flap pocket is responsible for substrate specificity, by binding to the unpaired 3' flap and by opening and kinking the DNA. Since the extrahelical 3'-flap pocket in archaeal Fen1 contains seven amino acids that are conserved to a great extent in human Fen1, we have mutated the four conserved or all seven amino acids in the human Fen1 extrahelical 3'-flap pocket to alanine. Our data suggest that the human extrahelical 3'-flap pocket mutants have lost substrate specificity to the double-flap DNA. Moreover, loss of high affinity for the unpaired 3'-flap suggests that the extrahelical 3'-flap pocket is essential for recognition and processing of the 'physiological' template. Human PCNA could stimulate the human Fen1 extrahelical 3'-flap pocket mutants but not restore their specificity. Thus the substrate specificity of Fen1 has been functionally conserved over a billion years from archaea to human. PMID- 15131256 TI - Chronic cardiac-specific thyrotoxicosis increases myocardial beta-adrenergic responsiveness. AB - Whereas many cardiac symptoms of thyrotoxicosis resemble those of the hyperadrenergic state, circulating catecholamines are reduced or normal in this condition. To test the hypothesis that the thyrotoxic heart is hypersensitive to catechol-amines, we studied beta-adrenergic signaling in a transgenic (TG) mouse in which the human type 2 iodothyronine deiodinase (D2) gene is expressed in myocardium. Because D2 converts T4 to T3, the active form of thyroid hormone, the D2 TG mouse exhibits mild, chronic thyrotoxicosis that is limited to the myocardium. In the current study, we determined that cAMP accumulation in response to either norepinephrine or forskolin treatment was increased in isolated ventricular myocardiocytes and membrane-enriched fractions prepared from these D2 TG hearts as compared with wild type. This increase in adenylyl cyclase (AC) Vmax could not be explained by changes in AC isoform expression or changes in the long or short forms of stimulatory G-protein Gsalpha, which were approximately 10% decreased in D2 TG membranes. However, Western analysis and ADP ribosylation studies suggest that the increase in AC Vmax is mediated by a decrease in the expression of inhibitory G proteins (Gialpha-3 and/or Goalpha). These data suggest that cardiac thyrotoxicosis leads to increased beta-adrenergic responsiveness of cardiomyocytes via alterations in the regulatory G-protein elements of the AC membrane complex. PMID- 15131257 TI - The protein kinase C signaling pathway regulates a molecular switch between transactivation and transrepression activity of the peroxisome proliferator activated receptor alpha. AB - Peroxisome proliferator-activated receptor (PPAR) alpha is a nuclear receptor implicated in several physiological processes such as lipid and lipoprotein metabolism, glucose homeostasis, and the inflammatory response. PPARalpha is activated by natural fatty acids and synthetic compounds like fibrates. PPARalpha activity has been shown to be modulated by its phosphorylation status. PPARalpha is phosphorylated by kinases such as the MAPKs and cAMP-activated protein kinase A. In this report, we show that protein kinase C (PKC) inhibition impairs ligand activated PPARalpha transcriptional activity. Furthermore, PKC inhibition decreases PPARalpha ligand-induction of its target genes including PPARalpha itself and carnitine palmitoyltransferase I. By contrast, PKC inhibition enhances PPARalpha transrepression properties as demonstrated using the fibrinogen-beta gene as model system. Finally, PKC inhibition decreases PPARalpha phosphorylation activity of hepatocyte cell extracts. In addition, PPARalpha purified protein is phosphorylated in vitro by recombinant PKCalpha and betaII. The replacement of serines 179 and 230 by alanine residues reduces the phosphorylation of the PPARalpha protein. The PPARalpha S179A-S230A protein displays an impaired ligand induced transactivation activity and an enhanced trans-repression activity. Altogether, our data indicate that the PKC signaling pathway acts as a molecular switch dissociating the transactivation and transrepression functions of PPARalpha, which involved phosphorylation of serines 179 and 230. PMID- 15131258 TI - Liver X receptors (LXRs) regulate apolipoprotein AIV-implications of the antiatherosclerotic effect of LXR agonists. AB - Liver X receptors (LXRs) regulate target genes that are critical in lipoprotein metabolism and atherosclerosis. Apolipoprotein AIV (ApoAIV) is an apolipoprotein that is associated with chylomicrons and high-density lipoproteins. Plasma ApoAIV level in humans is inversely correlated with coronary artery events and overexpression of ApoAIV in mice results in significant reduction in atherosclerosis. We report here that LXRs directly regulate apoAIV at the transcriptional level. Treatment of C57B6 mice with a synthetic LXR agonist, T0901317, resulted in significant increases in plasma apoAIV that was associated with high-density lipoprotein. Examination of both intestinal and liver apoAIV mRNA revealed specific increases in liver mRNA only. In a human heptoma HepG2 cell model, apoAIV mRNA was up-regulated upon the treatment with either native or synthetic LXR agonists. Nuclear run-on study revealed a significant increase in the ApoAIV transcriptional rate upon LXR activation. Examination of the human apoAIV proximal promoter revealed a potential LXR response element that demonstrated binding with HepG2 nuclear extracts. Cotransfection studies in HepG2 cells indicated that this responsive element was functional in mediating the human ApoAIV gene response to LXR agonists. In addition, we identified a functional LXR-responsive element at 3' end enhancer region of mouse ApoAIV gene. We conclude that ApoAIV is a direct target gene of LXRs that may contribute to the antiatherogenic effect of LXR activation. PMID- 15131259 TI - Transcription enhancer factor-5 and a GATA-like protein determine placental specific expression of the Type I human 3beta-hydroxysteroid dehydrogenase gene, HSD3B1. AB - The enzyme 3beta-hydroxysteroid dehydrogenase/isomerase (3betaHSD) is required for the biosynthesis of all active steroid hormones. It exists as multiple isoforms in humans and rodents, each a product of a distinct gene. Two isoforms, 3betaHSD I and II, are expressed in a tissue-specific manner in humans. 3betaHSD I is the only isoform expressed in the placenta, where it is required for the biosynthesis of progesterone and thus essential for the maintenance of pregnancy. We recently identified two transcription factors, activating protein-2gamma (AP 2gamma) and the homeodomain protein, distaless-3 (Dlx-3), that are expressed in both human and mouse trophoblast cells that were shown to be required for trophoblast-specific expression of the orthologous murine 3betaHSD, 3betaHSD VI. Although we identified specific binding sites for AP-2gamma and Dlx-3 in the distal promoter of the human 3betaHSD I gene, HSD3B1, it was found that these transcription factors were not involved in determining placental-specific expression of human 3betaHSD I. Instead, a 53-bp placental-specific enhancer element located between -2570 and -2518 of the HSD3B1 promoter was identified. Within this 53-bp element, two potential placental transcription factor binding sites were found. EMSAs with a 20-bp oligonucleotide containing these two potential placental-specific binding sites identified one of the binding sites specific for the transcription enhancer factor (TEF)-5, which is highly expressed in human placenta and in placental choriocarcinoma-derived JEG-3 cells and the other overlapping binding site, specific for a GATA-like protein. Site-specific mutations in either the TEF-5 binding site or in the GATA binding site, each resulted in complete loss of enhancer activity. The data indicate that TEF-5 and the GATA-like protein act in a coordinate manner to determine the placental specific expression of the human 3betaHSD I enzyme and therefore are critical for placental progesterone production required for the maintenance of pregnancy. PMID- 15131260 TI - The LATS2/KPM tumor suppressor is a negative regulator of the androgen receptor. AB - The androgen receptor (AR) is a member of the steroid receptor superfamily that plays critical roles in the development and maintenance of the male reproductive system and in prostate cancer. Actions of AR are controlled by interaction with several classes of coregulators. In this study, we have identified LATS2/KPM as a novel AR-interacting protein. Human LATS1 and LATS2 are tumor suppressors that are homologs of Drosophila warts/lats. The interaction surface of LATS2 is mapped to the central region of the protein, whereas the AR ligand binding domain is sufficient for this interaction. LATS2 functions as a modulator of AR by inhibiting androgen-regulated gene expression. The mechanism of LATS2-mediated repression of AR activity appears to involve the inhibition of AR NH2- and COOH terminal interaction. Chromatin immunoprecipitation assays in human prostate carcinoma cells reveal that LATS2 and AR are present in the protein complex that binds at the promoter and enhancer regions of prostate-specific antigen, and overexpression of LATS2 results in a reduction in androgen-induced expression of endogenous prostate-specific antigen mRNA. Immunohistochemistry shows that LATS2 and AR are localized within the prostate epithelium and that LATS2 expression is lower in human prostate tumor samples than in normal prostate. The results suggest that LATS2 may play a role in AR-mediated transcription and contribute to the development of prostate cancer. PMID- 15131261 TI - Paracrine regulation of ovarian granulosa cell differentiation by stanniocalcin (STC) 1: mediation through specific STC1 receptors. AB - Stanniocalcin (STC) in fish maintains calcium and phosphate homeostasis, whereas mammalian STC1 shows a diverse tissue expression pattern with ovary exhibiting the highest level. Based on the known expression of STC1 in theca/interstitial cells of the ovary, we generated recombinant N-glycosylated STC1 protein and tested its ability to modulate granulosa cell differentiation. In cultured rat granulosa cells obtained from early antral follicles, treatment with STC1 suppressed FSH-stimulated progesterone biosynthesis with minimal effects on estradiol and cAMP production. In mature granulosa cells, treatment with STC1 also suppressed human chorionic gonadotropin-induced progesterone production. The inhibitory effect of STC1 was accompanied by a pronounced suppression of the CYP11A transcripts and the FSH induction of functional LH receptors. In addition, STC1 was found to act downstream of adenyl cyclases in suppressing progesterone biosynthesis. We also tested the regulation of STC1 gene expression by gonadotropins. Treatment with pregnant mare serum gonadotropin decreased STC1 transcript levels in theca cells of maturing follicles, whereas subsequent treatment with human chorionic gonadotropin led to sustained suppression in the corpora lutea. Using radiolabeled recombinant STC1, receptor assays showed specific STC1 binding with a high affinity to granulosa cells. Because STC1 is expressed in ovarian theca/interstitial cells, the present demonstration of receptor binding and the specific actions of STC1 in granulosa cells suggest the existence of a follicular paracrine system in which theca cell-derived STC1 dampens the gonadotropin stimulation of granulosa cell differentiation. The observed STC1 suppression of progesterone, but not estradiol, production further suggests the potential role of this paracrine hormone as a luteinization inhibitor. PMID- 15131262 TI - Aberrant dynamics of histone deacetylation at the thyrotropin-releasing hormone gene in resistance to thyroid hormone. AB - Histone acetylation status influences transcriptional activity, and the mechanism of negative gene regulation by thyroid hormone remains unclear, although its impairment by a mutant thyroid hormone receptor (TR) is critical for resistance to thyroid hormone (RTH). We found a novel RTH mutant, F455S, that exhibited impaired repression of the TRH gene and had a strong dominant-negative effect on the gene. F455S strongly interacted with nuclear receptor corepressor (NCoR) and was hard to dissociate from it. To analyze the dynamics of histone acetylation status in vivo, we established cell lines stably expressing the TRH promoter and wild-type or F455S TR. Treatment with a histone deacetylase (HDAC) inhibitor completely abolished the repression of the gene by T3. The histones H3 and H4 at the TRH promoter were acetylated, and addition of T3 caused recruitment of HDACs 2 and 3 within 15 min, resulting in a transient deacetylation of the histone tails. TR and NCoR were located on the promoter, and T3 caused NCoR dissociation and steroid receptor coactivator-1 recruitment. In the presence of F455S, the histones were hyperacetylated, and HDAC recruitment and histone deacetylation were significantly impaired. This is the first report demonstrating the direct involvement of aberrant dynamics of chromatin modification in RTH. PMID- 15131263 TI - Origins of bilateral symmetry: Hox and dpp expression in a sea anemone. AB - Over 99% of modern animals are members of the evolutionary lineage Bilateria. The evolutionary success of Bilateria is credited partly to the origin of bilateral symmetry. Although animals of the phylum Cnidaria are not within the Bilateria, some representatives, such as the sea anemone Nematostella vectensis, exhibit bilateral symmetry. We show that Nematostella uses homologous genes to achieve bilateral symmetry: Multiple Hox genes are expressed in a staggered fashion along its primary body axis, and the transforming growth factor-beta gene decapentaplegic (dpp) is expressed in an asymmetric fashion about its secondary body axis. These data suggest that bilateral symmetry arose before the evolutionary split of Cnidaria and Bilateria. PMID- 15131264 TI - Regulation of an ATG7-beclin 1 program of autophagic cell death by caspase-8. AB - Caspases play a central role in apoptosis, a well-studied pathway of programmed cell death. Other programs of death potentially involving necrosis and autophagy may exist, but their relation to apoptosis and mechanisms of regulation remains unclear. We define a new molecular pathway in which activation of the receptor interacting protein (a serine-threonine kinase) and Jun amino-terminal kinase induced cell death with the morphology of autophagy. Autophagic death required the genes ATG7 and beclin 1 and was induced by caspase-8 inhibition. Clinical therapies involving caspase inhibitors may arrest apoptosis but also have the unanticipated effect of promoting autophagic cell death. PMID- 15131265 TI - Fluctuations and bistabilities on catalyst nanoparticles. AB - We show that coverage fluctuations on catalyst particles can drastically alter their macroscopic catalytic behavior. Scrutinizing the occurrence of kinetic bistabilities, it is demonstrated by molecular beam experiments on model catalysts that macroscopically observable bistabilities vanish completely with decreasing particle size, as previously predicted by theory. The effect is attributed to fluctuation-induced transitions between two kinetic reaction regimes, with a transition rate controlled by both particle size and surface defects. These results suggest that fluctuation-induced effects represent a general phenomenon affecting the reaction kinetics on nanostructured surfaces. PMID- 15131266 TI - Ultraconserved elements in the human genome. AB - There are 481 segments longer than 200 base pairs (bp) that are absolutely conserved (100% identity with no insertions or deletions) between orthologous regions of the human, rat, and mouse genomes. Nearly all of these segments are also conserved in the chicken and dog genomes, with an average of 95 and 99% identity, respectively. Many are also significantly conserved in fish. These ultraconserved elements of the human genome are most often located either overlapping exons in genes involved in RNA processing or in introns or nearby genes involved in the regulation of transcription and development. Along with more than 5000 sequences of over 100 bp that are absolutely conserved among the three sequenced mammals, these represent a class of genetic elements whose functions and evolutionary origins are yet to be determined, but which are more highly conserved between these species than are proteins and appear to be essential for the ontogeny of mammals and other vertebrates. PMID- 15131268 TI - An effective kinetic representation of fluctuation-driven neuronal networks with application to simple and complex cells in visual cortex. AB - A coarse-grained representation of neuronal network dynamics is developed in terms of kinetic equations, which are derived by a moment closure, directly from the original large-scale integrate-and-fire (I&F) network. This powerful kinetic theory captures the full dynamic range of neuronal networks, from the mean-driven limit (a limit such as the number of neurons N --> infinity, in which the fluctuations vanish) to the fluctuation-dominated limit (such as in small N networks). Comparison with full numerical simulations of the original I&F network establishes that the reduced dynamics is very accurate and numerically efficient over all dynamic ranges. Both analytical insights and scale-up of numerical representation can be achieved by this kinetic approach. Here, the theory is illustrated by a study of the dynamical properties of networks of various architectures, including excitatory and inhibitory neurons of both simple and complex type, which exhibit rich dynamic phenomena, such as, transitions to bistability and hysteresis, even in the presence of large fluctuations. The implication for possible connections between the structure of the bifurcations and the behavior of complex cells is discussed. Finally, I&F networks and kinetic theory are used to discuss orientation selectivity of complex cells for "ring model" architectures that characterize changes in the response of neurons located from near "orientation pinwheel centers" to far from them. PMID- 15131267 TI - Selective inhibition of calcineurin-NFAT signaling by blocking protein-protein interaction with small organic molecules. AB - Transient or reversible protein-protein interactions are commonly used to ensure efficient targeting of signaling enzymes to their cellular substrates. These interactions include direct binding to substrate, interaction with an accessory or scaffold protein, and positioning at subcellular locations in proximity to substrates. The existence of specialized targeting mechanisms raises the possibility of designing inhibitors that do not block enzyme activity per se, but rather interfere with targeting of the enzyme to one or more of its substrates within the cell. Here, we identify small organic molecules that specifically block targeting of the protein phosphatase calcineurin to its substrate nuclear factor of activated T cells (NFAT, also termed NFATc) and show that they are effective inhibitors of calcineurin-NFAT signaling. PMID- 15131269 TI - The UbcH8 ubiquitin E2 enzyme is also the E2 enzyme for ISG15, an IFN-alpha/beta induced ubiquitin-like protein. AB - Ubiquitin-(Ub) like proteins (Ubls) are conjugated to their targets by an enzymatic cascade involving an E1 activating enzyme, an E2 conjugating enzyme, and in some cases an E3 ligase. ISG15 is a Ubl that is conjugated to cellular proteins after IFN-alpha/beta stimulation. Although the E1 enzyme for ISG15 (Ube1L/E1(ISG15)) has been identified, the identities of the downstream components of the ISG15 conjugation cascade have remained elusive. Here we report the purification of an E2 enzyme for ISG15 and demonstrate that it is UbcH8, an E2 that also functions in Ub conjugation. In vitro assays with purified Ub E2 enzymes and in vivo RNA interference assays indicate that UbcH8 is a major E2 enzyme for ISG15 conjugation. These results indicate that the ISG15 conjugation pathway overlaps or converges with the Ub conjugation pathway at the level of a specific E2 enzyme. Furthermore, these results raise the possibility that the ISG15 conjugation pathway might use UbcH8-competent Ub ligases in vivo. As an initial test of this hypothesis, we have shown that a UbcH8-competent Ub ligase conjugates ISG15 to a specific target in vitro. These results challenge the concept that Ub and Ubl conjugation pathways are strictly parallel and nonoverlapping and have important implications for understanding the regulation and function of ISG15 conjugation in the IFN-alpha/beta response. PMID- 15131271 TI - The largest void on Earth? PMID- 15131270 TI - Consistency of a counterexample to Naimark's problem. AB - We construct a C*-algebra that has only one irreducible representation up to unitary equivalence but is not isomorphic to the algebra of compact operators on any Hilbert space. This answers an old question of Naimark. Our construction uses a combinatorial statement called the diamond principle, which is known to be consistent with but not provable from the standard axioms of set theory (assuming that these axioms are consistent). We prove that the statement "there exists a counterexample to Naimark's problem which is generated by aleph (1) elements" is undecidable in standard set theory. PMID- 15131272 TI - Epidemiology. Public enemy number one: tobacco or obesity? PMID- 15131273 TI - Global change. Getting warmer, however you measure it. PMID- 15131274 TI - U.S. research policy. NSF warned off Smithsonian pact. PMID- 15131275 TI - Fisheries science. Plan to count hatchery salmon criticized. PMID- 15131276 TI - National Science Foundation. State profiles join global review of science. PMID- 15131277 TI - Biodefense. Smallpox vaccines: looking beyond the next generation. PMID- 15131278 TI - Bird evolution. Surprise hummingbird fossil sets experts abuzz. PMID- 15131279 TI - Undergraduate science. Harvard joins reform movement. PMID- 15131280 TI - Neuroscience. Locating a new step in pain's pathway. PMID- 15131281 TI - Standards. A most unbearable weight. PMID- 15131282 TI - A Baghdad triptych. Reclaiming Iraq's past: life on the front lines. PMID- 15131283 TI - Huntington's disease. Unorthodox clinical trials meld science and care. PMID- 15131284 TI - American Association of Physical Anthropologists meeting. Tracking the evolutionary history of a "warrior" gene. PMID- 15131285 TI - American Association of Physical Anthropologists meeting. Chimpanzee gang warfare. PMID- 15131286 TI - American Association of Physical Anthropologists meeting. Seeing what an extinct monkey saw. PMID- 15131287 TI - Is human spaceflight now obsolete? PMID- 15131288 TI - Making a case for humans in space. PMID- 15131289 TI - How broad are water dimer bands? PMID- 15131290 TI - The problems of bonus pay. PMID- 15131292 TI - Beyond the ivory tower. "A vast machine": standards as social technology. PMID- 15131291 TI - Don't experiment on endangered ferrets! PMID- 15131294 TI - Physics. Colloids as big atoms. PMID- 15131293 TI - Neuroscience. Zif and the survival of memory. PMID- 15131295 TI - Ecology. Domains of diversity. PMID- 15131296 TI - Structural biology. The p75 NGF receptor exposed. PMID- 15131297 TI - Geophysics. A new paradigm for Earth's core-mantle boundary. PMID- 15131298 TI - Biophysics. Catching copper in the act. PMID- 15131299 TI - Lost at sea: where is all the plastic? PMID- 15131300 TI - Direct visual observation of thermal capillary waves. AB - We studied the free fluid-fluid interface in a phase-separated colloid-polymer dispersion with laser scanning confocal microscopy and directly observed thermally induced capillary waves at the interface in real space. Experimental results for static and dynamic correlation functions validate the capillary wave model down to almost the particle level. The ultralow interfacial tension, the capillary length, and the capillary time are found to be in agreement with independent measurements. Furthermore, we show that capillary waves induce the spontaneous breakup of thin liquid films and thus are of key importance in the process of droplet coalescence. PMID- 15131301 TI - Early Proterozoic ultrahigh pressure metamorphism: evidence from microdiamonds. AB - Microdiamonds from the Akluilak minette dykes (Nunavut, Canada) are similar to diamonds formed in subducted metamorphic rocks. High concentrations of unaggregated nitrogen and positive delta(15)N suggest that the microdiamonds formed within rocks subducted to ultrahigh pressures before being sampled by the minette magma 1.8 billion years ago. This ultrahigh pressure metamorphism in North America, probably related to the Trans-Hudson orogen (about 2 billion years ago), extends the occurrence of ultrahigh pressure metamorphism from 0.6 billion years to before 1.8 billion years ago and suggests that Phanerozoic-type subductions were active by the Early Proterozoic. PMID- 15131302 TI - Degradation of terrigenous dissolved organic carbon in the western Arctic Ocean. AB - The largest flux of terrigenous organic carbon into the ocean occurs in dissolved form by way of rivers. The fate of this material is enigmatic; there are numerous reports of conservative behavior over continental shelves, but the only knowledge we have about removal is that it occurs on long unknown time scales in the deep ocean. To investigate the removal process, we evaluated terrigenous dissolved organic carbon concentration gradients in the Beaufort Gyre of the western Arctic Ocean, which allowed us to observe the carbon's slow degradation. Using isotopic tracers of water-mass age, we determined that terrigenous dissolved organic carbon is mineralized with a half-life of 7.1 +/- 3.0 years, thus allowing only 21 to 32% of it to be exported to the North Atlantic Ocean. PMID- 15131303 TI - Old World fossil record of modern-type hummingbirds. AB - I report on tiny skeletons of stem-group hummingbirds from the early Oligocene of Germany that are of essentially modern appearance and exhibit morphological specializations toward nectarivory and hovering flight. These are the oldest fossils of modern-type hummingbirds, which had not previously been reported from the Old World. The findings demonstrate that early hummingbird evolution was not restricted to the New World. They further suggest that bird-flower coevolution dates back to the early Oligocene and open another view on the origin of ornithophily in Old World plants. PMID- 15131304 TI - Dioxygen binds end-on to mononuclear copper in a precatalytic enzyme complex. AB - Copper active sites play a major role in enzymatic activation of dioxygen. We trapped the copper-dioxygen complex in the enzyme peptidylglycine alphahydroxylating monooxygenase (PHM) by freezing protein crystals that had been soaked with a slow substrate and ascorbate in the presence of oxygen. The x-ray crystal structure of this precatalytic complex, determined to 1.85-angstrom resolution, shows that oxygen binds to one of the coppers in the enzyme with an end-on geometry. Given this structure, it is likely that dioxygen is directly involved in the electron transfer and hydrogen abstraction steps of the PHM reaction. These insights may apply to other copper oxygen-activating enzymes, such as dopamine beta-monooxygenase, and to the design of biomimetic complexes. PMID- 15131305 TI - Side-on copper-nitrosyl coordination by nitrite reductase. AB - A copper-nitrosyl intermediate forms during the catalytic cycle of nitrite reductase, the enzyme that mediates the committed step in bacterial denitrification. The crystal structure of a type 2 copper-nitrosyl complex of nitrite reductase reveals an unprecedented side-on binding mode in which the nitrogen and oxygen atoms are nearly equidistant from the copper cofactor. Comparison of this structure with a refined nitrite-bound crystal structure explains how coordination can change between copper-oxygen and copper-nitrogen during catalysis. The side-on copper-nitrosyl in nitrite reductase expands the possibilities for nitric oxide interactions in copper proteins such as superoxide dismutase and prions. PMID- 15131306 TI - Structure of nerve growth factor complexed with the shared neurotrophin receptor p75. AB - Neurotrophins are secreted growth factors critical for the development and maintenance of the vertebrate nervous system. Neurotrophins activate two types of cell surface receptors, the Trk receptor tyrosine kinases and the shared p75 neurotrophin receptor. We have determined the 2.4 A crystal structure of the prototypic neurotrophin, nerve growth factor (NGF), complexed with the extracellular domain of p75. Surprisingly, the complex is composed of an NGF homodimer asymmetrically bound to a single p75. p75 binds along the homodimeric interface of NGF, which disables NGF's symmetry-related second p75 binding site through an allosteric conformational change. Thus, neurotrophin signaling through p75 may occur by disassembly of p75 dimers and assembly of asymmetric 2:1 neurotrophin/p75 complexes, which could potentially engage a Trk receptor to form a trimolecular signaling complex. PMID- 15131307 TI - Periodic signaling controlled by an oscillatory circuit that includes protein kinases ERK2 and PKA. AB - Self-regulating systems often use robust oscillatory circuits. One such system controls the chemotactic signaling mechanism of Dictyostelium, where pulses of adenosine 3',5'-monophosphate (cAMP) are generated with a periodicity of 7 minutes. We have observed spontaneous oscillations in activation of the mitogen activated protein (MAP) kinase ERK2 that occur in phase with peaks of cAMP, and we show that ERK2 modulates cAMP levels through the phosphodiesterase RegA. Computer modeling and simulations of the underlying circuit faithfully account for the ability of the cells to spontaneously generate periodic pulses during specific stages of development. Similar oscillatory processes may occur in cells of many different species. PMID- 15131308 TI - Cognitive inflexibility after prefrontal serotonin depletion. AB - Serotonergic dysregulation within the prefrontal cortex (PFC) is implicated in many neuropsychiatric disorders, but the precise role of serotonin within the PFC is poorly understood. Using a serial discrimination reversal paradigm, we showed that upon reversal, selective serotonin depletion of the marmoset PFC produced perseverative responding to the previously rewarded stimulus without any significant effects on either retention of a discrimination learned preoperatively or acquisition of a novel discrimination postoperatively. These results highlight the importance of prefrontal serotonin in behavioral flexibility and are highly relevant to obsessive-compulsive disorder, schizophrenia, and the cognitive sequelae of drug abuse in which perseveration is prominent. PMID- 15131309 TI - The involvement of the anterior cingulate cortex in remote contextual fear memory. AB - Although the molecular, cellular, and systems mechanisms required for initial memory processing have been intensively investigated, those underlying permanent memory storage remain elusive. We present neuroanatomical, pharmacological, and genetic results demonstrating that the anterior cingulate cortex plays a critical role in remote memory for contextual fear conditioning. Imaging of activity dependent genes shows that the anterior cingulate is activated by remote memory and that this activation is impaired by a null alpha-CaMKII mutation that blocks remote memory. Accordingly, reversible inactivation of this structure in normal mice disrupts remote memory without affecting recent memory. PMID- 15131310 TI - GlyR alpha3: an essential target for spinal PGE2-mediated inflammatory pain sensitization. AB - Prostaglandin E2 (PGE2) is a crucial mediator of inflammatory pain sensitization. Here, we demonstrate that inhibition of a specific glycine receptor subtype (GlyR alpha3) by PGE2-induced receptor phosphorylation underlies central inflammatory pain sensitization. We show that GlyR alpha3 is distinctly expressed in superficial layers of the spinal cord dorsal horn. Mice deficient in GlyR alpha3 not only lack the inhibition of glycinergic neurotransmission by PGE2 seen in wild-type mice but also show a reduction in pain sensitization induced by spinal PGE2 injection or peripheral inflammation. Thus, GlyR alpha3 may provide a previously unrecognized molecular target in pain therapy. PMID- 15131312 TI - Stenting of symptomatic M1 stenosis of middle cerebral artery: an initial experience of 40 patients. AB - OBJECTIVE: To assess the safety and clinical efficacy of stenting for patients with symptomatic M1 stenosis of middle cerebral artery (MCA), and to assess the significance of classification based on location, morphology, and access of intracranial stenosis (LMA classification) in MCA stenting. METHODS: Forty patients with 42 symptomatic M1 stenoses refractory to medical therapy were enrolled in this study. The lesions were situated at M1 trunk (n=13), M1 origin (n=12), and M1 bifurcation (n=17), respectively, which were classified into type N (nonbifurcation lesions, n=13) and type A (prebifurcation, n=11), B (postbifurcation, n=14), C (lesion across the nonstenotic ostium of its branch, n=1), D (across the stenotic ostium of its branch, n=2), F (combinative lesions of prebifurcation and its small branch ostium, n=1) locations, morphologically into type A (n=15), B (n=23) and C (n=4) lesions, and into type I (mild-to moderate tortuosity and smooth access, n=17), II (severe tortuosity and/or irregular arterial wall, n=18), and III (excessively severe tortuosity, n=7) accesses. RESULTS: The technical successful rate was 97.6% for total lesions and 100%, 100%, and 85.7% for types I, II, and III accesses, respectively. The total complication rate was 10%. The mortality was 2.5% (1/40 patients), and 0%, 0%, and 25% for types A, B, and C lesions, respectively. During the median 10 months follow-up, there was no recurrence of transient ischemic attack or stroke in 38 available patients. Among 8 stenting vessels of seven patients with six-month follow-up angiography, 7 showed good patency and one showed restenosis. CONCLUSIONS: Stenting appears to be an effective and feasible therapy for symptomatic M1 stenoses, but also appears to have the higher periprocedural complications, which need strict procedural and periprocedural management to reduce the mortality and morbidity. The LMA classification seems to be helpful to work out the individual therapy and predict the results of stenting. A further study is needed to confirm the benefits of stenting of MCA stenosis. PMID- 15131313 TI - Autoantibodies against N-homocysteinylated proteins in humans: implications for atherosclerosis. AB - BACKGROUND AND PURPOSE: Homocysteine (Hcy)-thiolactone mediates protein N homocysteinylation in humans. Protein N-linked Hcy comprises a major pool of Hcy in human blood, greater that the "total" Hcy pool. N-homocysteinylated proteins are structurally different, compared with native proteins, and are thus likely to be recognized as neoself antigens and induce an autoimmune response. This study was undertaken to provide evidence for anti-Nepsilon-Hcy-Lys-protein antibody and to examine associations between the antibody level, Hcy, and stroke in humans. METHODS: ELISA was used to quantify anti-Nepsilon-Hcy-Lys-protein antibodies in human serum. RESULTS: We found that autoantibodies that specifically recognize Nepsilon-Hcy-Lys epitope on Hcy-containing proteins occur in humans. Serum levels of anti-Nepsilon-Hcy-Lys-protein autoantibodies positively correlate with plasma total Hcy levels, but not with plasma cysteine or methionine levels. In a group of exclusively male patients with stroke, mean level of anti-Nepsilon-Hcy-Lys protein autoantibodies was approximately 50% higher than in a group of healthy subjects. CONCLUSIONS: These findings support a hypothesis that Nepsilon-Hcy-Lys protein is a neoself antigen, which may contribute to immune activation, an important modulator of atherogenesis. PMID- 15131314 TI - Delayed decompressive surgery increases apparent diffusion coefficient and improves peri-infarct perfusion in rats with space-occupying cerebral infarction. AB - BACKGROUND AND PURPOSE: There is no conclusive experimental support that decompressive surgery in late stages of space-occupying cerebral infarction will improve outcome. We studied the effects of delayed decompressive surgery on the development of tissue damage, edema formation, and cerebral perfusion with different MRI techniques in a rat model of space-occupying cerebral infarction. METHODS: Permanent middle cerebral artery (MCA) occlusion was performed in 6 Fisher 344 rats. Decompressive surgery was performed 17 hours after the occlusion. Each animal was assessed before surgery and 2 and 4 hours after surgery by means, of diffusion-weighted T2-weighted, and flow-sensitive alternating inversion recovery perfusion-weighted MRI. Ischemic damage was also evaluated in hematoxylin-eosin-stained brain sections. RESULTS: Lesion volume as derived from apparent diffusion coefficient (ADC) maps decreased from 522+/-98 mm3 before to 405+/-100 mm3 (P=0.016) 4 hours after decompressive surgery, whereas lesion volume from T2 maps increased from 420+/-66 mm3 before to 510+/-92 mm3 (P=0.048) 4 hours after decompressive surgery. Midline shift decreased from 1.4+/-0.1 mm to 0.5+/-0.2 mm (P=0.001). Blood flow in the noninfarcted area of the ipsilateral hemisphere improved from 25+/-9 mL/min/100 g of tissue to 38+/-9 mL/min/100 g of tissue (P=0.035). Despite the pseudonormalization of ADC, irreversible damage was found in the entire MCA territory on histological evaluation. CONCLUSIONS: In rats with space-occupying cerebral infarction, delayed decompressive surgery leads to a decrease in lesion volume derived from ADC maps, which is probably because of an increase of extracellular water formation. There are no signs that this reflects rescue of ischemic tissue. PMID- 15131315 TI - Phase-contrast magnetic resonance imaging measurements of cerebral autoregulation with a breath-hold challenge: a feasibility study. AB - BACKGROUND AND PURPOSE: Vasomotor reactivity (VMR) testing can identify patients with hemodynamically critical cerebrovascular disease. The use of VMR has been limited by the invasiveness of most of the available methods and of acetazolamide as VMR stimulus. In the present study, we evaluated a completely noninvasive VMR approach by combining quantitative phase-contrast magnetic resonance imaging (MRI) with a breath-hold challenge. METHODS: Volume flow rates in the right and left internal carotid artery (ICA), basilar artery (BA), superior sagittal sinus, and sinus rectus were measured on 2-dimensional phase-contrast MR angiograms (MRAs) with a temporal resolution of 4.3 seconds. In 20 healthy control subjects, the VMR was assessed during 2 consecutive 30-second periods of breath-holding. RESULTS: A flow increase on breath-holding of 66% was found for the left ICA (240+/-54 mL/min to 398+/-120 mL/min; P<0.01), 59% for the right ICA (253+/-98 mL/min to 402+/-159 mL/min; P<0.01), 71% for the BA (107+/-48 mL/min to 184+/-79 mL/min; P<0.01), 62% for the superior sagittal sinus (232+/-75 mL/min to 375+/ 130 mL/min; P<0.01), and 65% for the sinus rectus (77+/-30 mL/min to 127+/-38 mL/min; P<0.01). The coefficient of variation for the total volume flow increase in the brain feeding arteries (ICAs and BA) between the first and the second breath-holds was 18%. CONCLUSIONS: The combination of MRA phase-contrast volume flow measurements and a breath-holding challenge allows for a fast, completely noninvasive, and reproducible assessment of VMR. PMID- 15131316 TI - Lower serum triglyceride level is associated with increased stroke severity. AB - BACKGROUND AND PURPOSE: A previous study showed that low triglyceride concentration predicts higher mortality after stroke. The aim of our study was to determine whether serum triglyceride level is associated with stroke severity on admission. METHODS: 863 consecutive patients with acute ischemic stroke were included. Serum triglyceride level was measured within 36 hours after stroke onset. Stroke severity on admission was assessed using Scandinavian Stroke Scale (SSS). The patients were divided into 2 groups: those with severe stroke (SSS < or =25) and those with mild/moderate stroke (SSS >25). RESULTS: Patients with severe stroke had significantly lower serum triglyceride level than patients with mild/moderate stroke (1.4+/-0.6 versus 1.7+/-1.3 mmol/L). After adjusting for age, sex, atrial fibrillation, diabetes mellitus, obesity, and ischemic heart disease, patients with triglyceride >2.3 mmol/L had lower risk of severe stroke than those with triglyceride < or =2.3 mmol/L (OR: 0.58; 95% CI: 0.35 to 0.95). CONCLUSIONS: Our results suggest that lower level of triglyceride is associated with the more severe stroke. PMID- 15131317 TI - Influence of maternal stress on metal-induced pre- and postnatal effects in mammals: a review. AB - Studies in rodents have shown that, during pregnancy, maternal stress from restraint, noise, light, and heat among other factors may be associated with adverse effects on embryo/fetal and postnatal development. Moreover, it is also well known that exposure to certain metal levels during gestation can also cause maternal and developmental toxicity. Because potentially, pregnant women may be concurrently exposed to metals and various types of stress, the influence of maternal stress on the metal-induced adverse pre- and postnatal effects has been investigated for a number of elements. This influence is reviewed here. It is concluded that maternal stress enhances the metal-induced embryo/fetal and developmental toxicity only at doses of the metal which are also clearly toxic to the dam. PMID- 15131318 TI - Effects of zinc on cell-mediated immunity in chronic hemodialysis patients. AB - Thirteen healthy subjects and 20 hemodialysis patients were studied to observe the delayed hypersensitivity skin tests (DHSTs) and phytohemagglutinin (PHA) stimulating lymphocyte blastogenesis. Significant differences were observed between the groups. Controls had a higher proportion of positive skin reaction than hemodialysis patients in relation to Escherichia coli (p<0.01) and tuberculin (PPD) (p<0.05). Regarding lymphocyte blastogenesis stimulated by phytohemagglutinin (PHA), cell proliferation was more accentuated in controls than hemodialysis patients (p<0.05). On the other hand, serum zinc was elevated in controls (78 +/- 8 microg/dL) in comparison to hemodialysis patients (71 +/- 33 microg/dL) (p<0.05). Of the 20 hemodialysis patients, 8 patients were maintained on long-term hemodialysis before and after zinc therapy, with the aim of studying DHST and PHA-stimulating lymphocyte blastogenesis. There was a significant improvement of DHST response to E. coli antigen after 100 d of zinc treatment (p<0.01), and with the discontinuation of therapy, the DHST responses decreased back to the initial values (p<0.05). Zinc administration also increased the lymphocyte proliferation induced by PHA from 31386 +/- 3974 to 42480 +/- 5242 cpm (mean +/- SD) (p<0.05). These results indicated that zinc therapy improved in vivo and in vitro DHST and lymphocyte function of hemodialysis patients and that its discontinuation suppressed all of the benefits observed. PMID- 15131319 TI - Antioxidant superoxide dismutase activity in obese children. AB - The aim of the study was to evaluate the antioxidative Cu/Zn-SOD (superoxide dismutase) response to obesity-related stress in obese children compared to a similar-aged control group. Forty-eight exogenic obese children and 11 healthy children were compared for red cell Cu/Zn-SOD, glucose, and lipid profiles and the relations between them were investigated. Antioxidant response as Cu/Zn-SOD was significantly higher in the obese group (p<0.05). Although glucose and lipid levels were statistically higher in the obese group, a certain relation with the SOD level was not established in childhood. This is the first study showing the oxidative stress caused by obesity and related antioxidative response even in the childhood period. Interventions, including diet modifications, should be kept in mind to diminish the obesity-related oxidative stress from the childhood period. PMID- 15131320 TI - Age-related changes of elements with their relationships in human cranial and spinal nerves. AB - To elucidate compositional changes of peripheral nerves with aging, the authors investigated age-related changes of elements and their relationships in the optic, trigeminal, vagus, median, radial, ulnar, femoral, sciatic, tibial, and common peroneal nerves by inductively coupled plasma-atomic emission spectrometry. The subjects consisted of 10 men and 12 women, ranging in age from 65 to 91 yr. It was found that although accumulations of Ca and P occurred only in the trigeminal nerve at old age, it hardly occurred in the optic, vagus, median, radial, ulnar, femoral, sciatic, tibial, and common peroneal nerves at old age. The average contents of Ca and P were three and two times higher in the trigeminal nerve than in the other nine kinds of nerve, respectively. Likewise, the average content of Mg was a little higher in the trigeminal nerve compared with the other nerves. With regard to the relationships among elements, significant direct correlations were found among the contents of Ca, P, S, and Mg in most, but not all, 10 kinds of nerve. In the trigeminal nerve, a significant inverse correlation was found between the contents of S and the other elements, such as Ca, P, and Mg. Regarding the relationships between the contents of S and other elements, the nerves, except for the trigeminal nerve, differed from those found in the arteries previously reported. PMID- 15131321 TI - Antagonistic effect of scutellarin on the toxicity of selenium in rat livers. AB - Selenium has both nutritional function and toxicity according to its concentration and species. To counteract the toxicity of selenium, scutellarin was investigated. Wistar rats were supplemented with 40 microg Se/kg/d as sodium selenite, 40 microg Se/kg/d with 20 mg/kg/d scutellarin, and 20 mg/kg/d scutellarin, respectively, for 15 d. The mRNA levels and activities of glutathione peroxidase (GSH-Px) and thioredoxin reductase (TR), and the malondialdehyde (MDA) contents were measured. Reactive oxygen species (ROS) were detected by chemiluminescence assay, and tissue conformation was investigated by histological study. The results showed significant decreases of mRNA levels and activities of GSH-Px and TR and a significant increase of MDA content in livers of the Se-treated rats (p<0.05, compared with the control). Supplementation of scutellarin to the Se-treated group significantly inhibited the decreases of mRNA levels and activities, and the increase of MDA content (p<0.05, compared with the Se-treated group). Meanwhile, scutellarin-scavenged ROS generated in the mixture of sodium selenite, reduced glutathione, and oxygen. Liver injury was displayed in slices exposed to selenium at the present dose. The groups treated with both selenium and scutellarin or only scutellarin did not show significant tissue damage. Thus, scutellarin had an antagonistic effect against the toxicity of selenium. PMID- 15131322 TI - Concentrations of copper, iron, and zinc in the major organs of the wistar albino and wild black rats: a comparative study. AB - The concentrations of copper, iron, and zinc in the major organs of Wistar albino (Rattus norvegicus) and wild black rats (Rattus rattus) were measured by means of atomic absorption spectroscopy. The copper levels in the kidneys and liver of the Wistar albino rats (WARs) were significantly higher (p<0.05) than in the wild black rats (WBRs). There were no significant differences in the concentrations of zinc in the liver, lungs, kidneys, and brain between the two study groups, but zinc was significantly higher in the spleen (p<0.05) and lower in the heart (p<0.05) of WAR, compared to WBRs. Iron was significantly higher (p<0.05) in the heart and spleen of WBRs, compared to WARs. There were no extreme differences in the organ concentrations of trace elements between the two species, but, cumulatively, the WARs tend to have higher metallic concentrations in their system than the WBRs. The potential of these differences on the experimental results should not be overlooked and will serve as basis to further consider the complex interrelationships of these animals in their microenvironments and macroenvironments. PMID- 15131323 TI - Metallic tin is an adjuvant for anaphylaxis in rats. AB - Metallic tin powder is known to have an adjuvant-like property, by which it increases the levels of natural antibodies and induced hemagglutinins, and it enhances the induction of allergic encephalomyelitis in rats. In the present work, metallic tin is shown to be an adjuvant for a different immunologic process, anaphylactic sensitization. The new data support the notion that metallic tin causes polyclonal B-cell activation with proliferation of plasma cells. PMID- 15131324 TI - Mechanistic dissection of pain: from DNA to animal models. PMID- 15131325 TI - Assessment of acute thermal nociception in laboratory animals. AB - Models of acute nociception using a thermal stimulus are widely employed as screening methods for nociceptive properties of new drug compounds. In this chapter, detailed descriptions for conducting of two of the most commonly used models; the hot plate test and the "Hargreaves test," are described. These models are applicable to both rats and mice and have the advantage of allowing repeated and multiple testing using a single animal because the stimulus is transitory and produces no tissue damage. Additionally, a modification of these models using a skin-twitch reflex that is applicable to large laboratory animals such a dogs or sheep is described. Guidance concerning potential confounding variable are discussed, as are tips for reducing variably among testing sessions. PMID- 15131326 TI - Tissue injury models of persistent nociception in rats. AB - The purpose of this chapter is to provide guidance to the novice investigator as to two models of ongoing nociception in rats. The models described herein are the formalin test, in which an irritant is injected subcutaneously into a dorsal paw and the numbers of flinches produced over 60 min are counted, and a mild burn model that produces a transitory primary and secondary thermal and mechanical hyperalgesia lasting approx 90 min. These models allow assessment of spinal sensitization, which may be an important factor when considering plasticity associated with human pain states. Detailed protocols using both manual and automated counting for the formalin test are included, as are methods concerning data analysis. PMID- 15131327 TI - Segmental spinal nerve ligation model of neuropathic pain. AB - Since its introduction in 1992, the spinal nerve ligation (SNL) model of neuropathic pain has been widely used for various investigative works on neuropathic pain mechanisms as well as in screening tests for the development of new analgesic drugs. This model was developed by tightly ligating one (L5) or two (L5 and L6) segmental spinal nerves in the rat. The operation results in long lasting behavioral signs of mechanical allodynia, heat hyperalgesia, cold allodynia, and ongoing pain. In the process of widespread usage, however, many different variations of the SNL model have been produced, either intentionally or unintentionally, by different investigators. Although the factors that cause these variations themselves are interesting and important topics to be studied, the pain mechanisms involved in these variations are likely different from the original model. Therefore, this chapter describes, in detail, the method for producing the spinal nerve ligation model that will minimally induce potential factors that may contribute to these variations. It is hoped that this description will help many investigators to produce a consistent animal model with uniform pathophysiological mechanisms. PMID- 15131328 TI - Partial sciatic nerve transection. AB - Partial sciatic nerve transection (PST) of the sciatic nerve of rats and mice is described as a model of painful neuropathy. The rationale for developing this model was to establish a simple partial nerve injury without application of foreign material. In contrast to the frequently used model of chronic constriction injury (CCI), PST allows to relate animal behavior and drug effects to endoneurial changes, undisturbed by major epineurial inflammation. PST is easy to perform in rats and mice and leads to reproducible pain-related behavior for 5 wk or longer. PMID- 15131329 TI - Modeling diabetic sensory neuropathy in rats. AB - The procedures to induce insulin-deficient diabetes in rats using streptozotocin are described along with a number of insulin treatment regimes that can be used to maintain these animals at different degrees of glycemia for periods of weeks to months. Streptozotocin-diabetic rats develop tactile allodynia, hyperalgesia following paw formalin injection and abnormal responses to thermal stimulation and the detailed methods used to evaluate these behavioral indices of abnormal sensory function are provided. PMID- 15131330 TI - Sciatic inflammatory neuropathy in the rat: surgical procedures, induction of inflammation, and behavioral testing. AB - Peripheral nerve damage involves inflammation, and is frequently causal to the development of neuropathic pain. However, inflammatory neuropathies often occur in the absence of trauma. We have recently developed an animal model of neuropathic pain where allodynia is induced by nerve inflammation rather than injury. This sciatic inflammatory neuropathy (SIN) model was developed to understand immunologic, neuropathic, and spinal mechanisms underlying allodynia in the territory of the sciatic nerve as well as in extraterritorial and contralateral ("mirror image") sites. A specially designed indwelling catheter system allows immune activators to be selectively injected around one healthy sciatic nerve in awake, behaving rats. Here, we provide detailed procedures on the construction and implantation of chronic indwelling perisciatic catheters used to create SIN. Detailed procedures for implantation of intrathecal catheters via a lumbar vertebra 5 and 6 approach in the same rat are also provided. Methods for testing allodynia and for data analysis are additionally described so to provide all the steps needed for behavioral experimentation. PMID- 15131331 TI - A rat pain model of vincristine-induced neuropathy. AB - Vincristine belongs to the family of vinca alkaloids used for treatment of malignant tumors. Clinical application of these agents is often associated with dose-dependent painful neuropathy due to damages to the peripheral axons. A rat model of vincristine-induced hyperalgesia was developed through intravenous injection of vincristine by Aley et al. (1996) and was later modified by Nozaki Taguchi et al. (2001) using continuous intravenous infusion of vincristine. This model provides consistent and long-lasting neuropathic pain states mimicking vincristine-induced pain conditions in human patients. Therefore, this model is a valuable means of studying the mechanisms and pharmacology of vincristine-induced neuropathic pain. In this chapter we describe in detail steps the generation of vincristine-induced neuropathy in rats through continuous intravenous infusion of vincristine. PMID- 15131332 TI - Loop dialysis catheter: a technology for chronic spinal dialysis in a freely moving rat. AB - To permit long-term measurement of time-dependent changes in levels of dialyzable drugs and transmitters in the spinal intrathecal (IT) space of the unanesthetized rat, we developed a dialysis catheter for chronic placement. This was accomplished by constructing a loop-flexible probe 9 cm in length from PE-5 polyethylene tubing and 4 cm dialysis membrane. This loop catheter was inserted through an incision in the cisternal membrane and passed to the lumbar enlargement. The ends of the catheter were then externalized on the top of the head. For dialysis, an external end of the loop catheter was connected to a syringe pump and perfused with artificial cerebrospinal fluid (CSF) and the out flow collected. Using this system we demonstrate: a) a significant increase in spinal CSF concentrations of amino acids after spinal traumatic injury, b) the permeability of a drug (methylprednisolone) through the blood-brain barrier into the spinal extracellular space after systemic delivery, c) effect of IT injection of N-methyl-D-aspartate on secondary amino acid release, and c) clearance of sufentanyl after IT delivery and corresponding development of thermal nociception. The loop dialysis catheter provides a robust experimental tool for studying time-dependent changes in the concentration of diffusible substances in spinal CSF over an extended postimplantation interval and allows comparison of these changes with concurrently assessed behavioral indices. PMID- 15131333 TI - Intrathecal catheterization and drug delivery in the rat. AB - The spinal cord represents a complex system that serves in the encoding of sensory information and organization of autonomic and somatomotor outflow. As such, it has become a target of investigation for subjects ranging from pain to hypertension to motor spasticity. A primary method in such investigations is the specific delivery of drugs into the spinal intrathecal space to assess such agents with the actions limited to the spinal cord. The rat has served as a primary model in these investigations. It provides for the ability to deliver such agents both acutely and chronically in the absence of anesthesia or restraint. These goals can be accomplished by the placement of intrathecal catheters in the spinal space. This model, first demonstrating the feasibility of routinely catheterizing the lumbar intrathecal space in 1976, led directly to enabling a large number of studies focusing on spinal drug actions. As a test model, it has provided for the acquisition of an exceptional amount of information on the pharmacological and physiological mechanisms of spinal function, drug screening, drug efficacy and safety studies, and pharmacokinteics of spinally delivered drugs. The modified surgical method of intrathecal catheter placement, microinjection drug delivery, and behavioral parameters are described in detail. PMID- 15131334 TI - Trigeminal neuronal recording in animal models of orofacial pain. AB - The electrical signal associated with nerve cells, mainly as a result of changes in the membrane potential during functional activity, can be recorded extracellularly to study central mechanisms underlying sensory processing. The secondary neurons in the spinal trigeminal complex receive inputs from peripheral neurons that innervate the orofacial region and forward information to the higher levels of the nervous system. Analyzing activity patterns of trigeminal neurons related to pain perception has proven to be an efficient method in studying orofacial pain mechanisms. Here we describe some basic techniques and tips for extracellular single neuron recording from the subnucleus caudalis of the trigeminal spinal nucleus in rats with orofacial injury. Two different rat models with temporomandibular joint inflammation and inferior alveolar nerve transection are described. PMID- 15131335 TI - In vivo electrophysiology of dorsal-horn neurons. AB - The dorsal horn of the spinal cord is a key relay in the transmission of sensory information to the brain. Furthermore, this circuitry of spinal-cord neurons, and hence the spinal processing of sensory information, is subject to a great deal of plasticity, both pharmacological and physiological, in persistent pain states. This chapter describes in detail the procedure by which the activity and pharmacological modulation of these dorsal-horn neurons can be recorded in vivo in anesthetized rats, allowing a comprehensive study of spinal sensory processing in an intact and integrated system. The chapter covers the surgical preparation of the animal for electrophysiological recording; isolating and recording the activity of a single dorsal-horn neuron; and identifying the type of dorsal-horn neuron recorded by characterizing the neuronal response to a variety of peripheral stimuli. The study of these neuronal responses in a variety of persistent pain states, such as carrageenan-induced inflammation and neuropathy induced by L5/L6 spinal nerve ligation, together with the study of their pharmacological modulation by locally or systemically administered drugs, is also described. PMID- 15131336 TI - Single-fiber recording: in vivo and in vitro preparations. AB - This chapter focuses on in vivo and in vitro recording setups of extracellular single-unit recordings of peripheral sensory nerve or dorsal root fibers in rodents. Extracellular single-unit recording methods have been used to obtain a wealth of data about the properties of peripheral nervous system (PNS) and central nervous system (CNS) structures. The rationale for studying the activity of single-unit primary afferent fibers is predicated on the significance of relatively fine variations of fiber responsiveness to mechanical, chemical, and/or thermal stimuli. It involves microdissection of nerve fiber bundles until the electrical activity of a single fiber is isolated. Electrophysiological changes in thresholds and discharge rates of peripheral nociceptors to polymodal stimuli can provide neurophysiological correlation to behavioral hyperalgesia and allodynia as well as to cellular differences observable with immunohistochemistry. This chapter gives an overview about the necessary general and special equipment, details about the different setups and tissue preparations. Additionally, the chapter informs about the procedure of recording from single units, data acquisition and analysis including unit isolation criteria and techniques for spike discrimination techniques and fiber classification. It describes criteria for the classification of nociceptors and identification of cutaneous afferent units. PMID- 15131337 TI - Anatomical identification of neurons responsive to nociceptive stimuli. AB - We describe methods for labeling and identifying neurons within the central nervous system involved in the transmission of nociceptive stimuli. The most reliable methods are physiological identification followed by intracellular injection or immunocytochemical detection of stimulus-induced markers such as Fos. These latter strategies are used with appropriate controls to distinguish neurons activated secondarily (e.g., motor response or inhibitory neurons) by the nociceptive stimuli. Other methods include location and morphology as determined by standard cytological and tracing methods and/or the presence of specific neurochemical markers such as substance P determined by immunocytochemistry. PMID- 15131338 TI - Isolation and culture of sensory neurons from the dorsal-root ganglia of embryonic or adult rats. AB - There is increasing use of isolated sensory neuronal preparations to examine the cellular mechanisms involved in pain signaling. Indeed, these in viro preparations have several advantages that make them beneficial for examining physiological and/or pathological processes affecting neuronal function. With isolated cells it can be determined whether various inflammatory mediators and algogenic agents have direct actions on sensory neurons. Additionally, the intracellular signaling pathways for agents that modulate the excitability and sensitization of sensory neurons can be examined. Finally, the concentrations of mediators and drugs that are used to alter cell function can be well controlled. The purpose of this chapter is to provide the reader with detailed methods for the harvest and growth of embryonic and adult rat sensory neurons (dorsal root ganglia neurons) in culture. Because numerous methods for growing sensory neurons exist, the rationale for certain aspects of the protocols described in the chapter are included, as are discussions of potential pitfalls. PMID- 15131339 TI - Primary cultures of neonatal rat spinal cord. AB - Primary cultures of neurons provide opportunities to study the cell biology of neurons under controlled conditions. Because differences exist in cellular properties among populations of neurons in the brain, survival requirements for neurons among these regions differ as well. This chapter outlines protocols for the preparation of primary cultures of spinal cord from 2-d-old neonatal rats. One protocol prepares cultures enriched in neurons and an alternative procedure prepares cultures enriched in non-neuronal cells. Comparison of biochemical data between these two culture preparations allows deductions of effects of treatments on neurons in the cultures. Limitations in interpretation of data obtained from cultured neurons are discussed. PMID- 15131340 TI - Single-cell laser-capture microdissection and RNA amplification. AB - Generating gene-expression profiles from laser-captured cells requires the successful combination of laser-capture microdissection, RNA extraction, RNA amplification, and microarray analysis. To permit single-cell gene-expression profiling, the RNA amplification method has to be sufficiently powerful to bridge the gap between the amount of RNA available from a single cell to what is required by the microarray, a gap that spans 5 to 6 orders of magnitude. This chapter focuses on the amplification of RNA using a two-round T7 RNA amplification method. The protocols described are adapted for laser-captured material and have been used to generate gene expression profiles from single laser-captured cells. PMID- 15131341 TI - Semi-quantitative real-time PCR for pain research. AB - In this chapter we cover what we have found to be a "best practice" for real-time polymerase chain reaction (PCR) for relative mRNA quantification. We describe our techniques for tissue-sample collection and freezing, sample handling for quick and reproducible extraction of total RNA, first strand cDNA synthesis, real-time PCR amplification, and template dilution and storage for PCR. We offer our insights on intron-spanning primer design for genes (when applicable), effective primer selection vs reaction optimization, and relative quantification and sample normalization using housekeeping genes. Comments are also provided on the choice of PCR reagents including fluorescent probes, prevention of PCR "carry over," and on the practical aspects of real-time PCR theory and interpretation. PMID- 15131342 TI - Functional genomic analysis in pain research using hybridization arrays. AB - Hybridization array technology makes it possible to compare global gene expression patterns in any experimental context for which good-quality RNA can be generated. To date, DNA arrays have been used as a tool to compare functional genomic changes (differences in wholesale gene expression) in studies that cover an impressive variety of research disciplines including cancer, yeast genomics, and, more recently, neuroscience and behavior. The basic premise of the array experiment is that one interrogates a panel of probes (gene-specific cDNA fragments or gene-specific oligonucleotides that have been immobilized on a solid support) with RNAs (targets) from control and treated experimental samples that have been either radioactively or fluorescently labeled. Signal derived from either competitive (both samples on a single chip) or differential (one sample/one chip) hybridization is used to calculate relative gene expression. There are three widely used platforms available to perform array experiments (Affymetrix GeneChips, oligonucleotide arrays, and membrane-based cDNA arrays) and each platform offers advantages and limitations. The experimental description in this chapter explains, in detail, how to perform a hybridization array using the macroarray platform. PMID- 15131343 TI - Generation of transgenic mice. AB - In this post-genomic era, emphasis has shifted from identifying genes to understanding the physiological functions of gene products and their implications in human diseases. The use of transgenic mice is one of the key approaches in elucidating gene function and regulation. Transgenic mice have wide applications in biomedical research. These include (1) gain-of-function studies by overexpressing a protein in a tissue-specific manner; (2) loss-of-function studies by overexpressing a dominant-negative construct; (3) mapping functional domains by expressing various mutated constructs in a null (knockout mice) background; (4) mapping regulatory elements using a reporter gene; and (5) labeling subsets of cells with fluorescent proteins for live in vivo imaging of cell structure and function. In this chapter, we provide overviews as well as detailed protocols for each step involved in the generation of transgenic mice, including the selection of regulatory elements, purification of DNA, obtaining fertilized eggs, pronuclear injection, and genotyping of transgenic mice. PMID- 15131344 TI - Knockout mouse models in pain research. AB - Gene targeting in mice by homologous recombination is a powerful approach to study the role of specific genes in vivo. This technology is now applied to pain related genes to understand molecular mechanisms of nociceptive behaviors. In this chapter, we provide detailed methodological information for the construction of knockout animals, exemplified by the generation of mice lacking opioid receptor genes. We report our protocols for the production, maintenance, transfection, and selection of embryonic stem (ES) cells, as well as for blastocyst injection, which are generally applicable to any gene-targeting project. We also describe strategies for the construction of targeting vectors, as well as for ES cell and animal genotyping, in the context of mu, delta, and kappa opioid receptor genes. We finally provide a few examples of mouse phenotyping in pain behavioral assays. PMID- 15131345 TI - Transgene inheritance in plants. AB - The patterns of transgene inheritance in plants and the possible explanations for non-Mendelian transmission are reviewed. The non-Mendelian inheritance of a transgene has been recorded with a frequency between 10% and 50% in transgenic plants produced either by Agrobacterium-mediated transformation or through particle bombardment. Different effects such as deletion, duplication, rearrangement, repeated sequence recombination as well as gene interaction have been observed for transgenic loci. The nature of the recipient genome, nature of the transgene and the interactions between them seem to contribute to the non Mendelian segregation of transgenes. PMID- 15131346 TI - Molecular markers in breeding for virus resistance in barley. AB - The soil-borne barley yellow mosaic virus disease (BaMMV, BaYMV, BaYMV-2) and the aphid-transmitted barley yellow dwarf virus (BYDV) are serious threats to winter barley cultivation. Resistance to barley yellow mosaic virus disease has been identified in extensive screening programmes and several recessive resistance genes have been mapped, e.g. rym4, rym5, rym9, rym11, rym13. In contrast to barley yellow mosaic virus disease, no complete resistance to BYDV is known in the barley gene pool, but tolerant accessions have been identified and QTL for BYDV-tolerance have been detected on chromosomes 2HL and 3HL. The use of resistance and tolerance in barley breeding can be considerably improved today by molecular markers (RFLPs, RAPDs, AFLPs, SSRs, STSs, SNPs), as they facilitate (i) efficient genotyping and estimation of genetic diversity; (ii) reliable selection on a single plant level independent of symptom expression in the field (iii) acceleration of back crossing procedures; (iv) pyramiding of resistance genes; (v) detection of QTL and marker-based combination of positive alleles; and (vi) isolation of resistance genes via map-based cloning. PMID- 15131347 TI - Fluorescence-based AFLPs occur as the most suitable marker system for oilseed rape cultivar identification. AB - Three different types of molecular markers, RAPD, SSR and fluorescence-based AFLP, were evaluated and compared for their ability to identify oilseed rape cultivars. The direct comparison of RAPD, SSR and AFLP approaches in cultivar identification showed that the AFLP methodology detected polymorphisms more efficiently than either RAPD or SSR methods. For the characterisation of six oilseed rape cultivars, 60 RAPD primers were tested and only eight of them (14%) detected sufficient levels of polymorphism. Five microsatellites out of fifteen tested were polymorphic, but in all loci, except one, only two different alleles were detected. This result indicated the limited degree of polymorphism found in Brassica napus. Each of the six tested AFLP combinations detected polymorphisms, the best combination (M-CAA/E-ACT) had 26% polymorphic peaks from a total of 90 peaks and could distinguish the analysed cultivars and 4 out of 5 core lines of cultivars. The results presented show that florescence-based AFLP is, for the purposes of oilseed rape cultivar fingerprinting, a more suitable approach than either RAPD or SSR. PMID- 15131348 TI - The influence of morphological differences between sunflower inbred lines on their SCA effects for yield components. AB - Genetic distances between seven oil sunflower inbred lines expressed in the variability of five morphological traits were analysed in terms of their effect on the specific combining ability (SCA) effects in these lines for yield component traits. The multivariate analysis of variance (MANOVA) and other related multidimensional methods were used to recognize the differences between genotypes with respect to morphological traits. Canonical analysis was applied for a graphical configuration of parental lines and diallele hybrids in the two dimensional space. As a measure of the difference between parental lines with respect to the analysed morphological traits jointly, i.e. as a measure of the morphological distance between these forms, Mahalanobis' distance was used. Phenotypic differences between lines were estimated using Mahalanobis' distance for each pair of crossed lines calculated for all the analysed morphological traits. The effect of morphological differences between inbred lines on the mean SCA effect for individual yield components, measured by the linear regression, was significant for oil yield per plant and for the number of seeds per plant. PMID- 15131349 TI - DNA polymorphism among barley NILs of cv. Pallas, carrying genes for resistance to powdery mildew (Blumeria graminis f. sp. hordei). AB - Barley powdery mildew, caused by the pathogen Blumeria graminis f. sp. hordei is an important disease of barley (Hordeum vulgare L.). The random amplified polymorphic DNA (RAPD) method was used to detect DNA polymorphism among 7 Pallas near-isogenic lines (NILs) carrying Mla3, Mla12, Mlk, Mlp, Mlat, Mlg and MlLa genes for resistance to B. graminis f. sp. hordei. From among 500 random 10-mer primers tested, 3 were specific for NIL P2 (Mla3), 1 for P10 (Mla12), 6 for P17 (Mlk), 46 for P19 (Mlp), 4 for P20 (Mlat), 6 for P21 (Mlg), and 4 for P23 (MlLa). The results of this study demonstrated that the RAPD technique is a useful tool for detecting DNA polymorphism among Pallas NILs. PMID- 15131350 TI - Localisation of rDNA in the Lupinus genome during the cell cycle. AB - Observations of a specific rDNA locus behaviour during the cell cycle were made by fluorescent in situ hybridisation (FISH) in 12 Lupinus species. Due to the pattern of chromatin de-condensation in that locus, the number of relevant sites in interphase nuclei was twice as high as the number of signals on metaphase chromosomes. The description of successive phases and an attempt of an explanation are given. PMID- 15131351 TI - The dog genome map and its use in mammalian comparative genomics. AB - The dog genome organization was extensively studied in the last ten years. The most important achievements are the well-developed marker genome maps, including over 3200 marker loci, and a survey of the DNA genome sequence. This knowledge, along with the most advanced map of the human genome, turned out to be very useful in comparative genomic studies. On the one hand, it has promoted the development of marker genome maps of other species of the family Canidae (red fox, arctic fox, Chinese raccoon dog) as well as studies on the evolution of their karyotype. But the most important approach is the comparative analysis of human and canine hereditary diseases. At present, causative gene mutations are known for 30 canine hereditary diseases. A majority of them have human counterparts with similar clinical and molecular features. Studies on identification of genes having a major impact on some multifactorial diseases (hip dysplasia, epilepsy) and cancers (multifocal renal cystadenocarcinoma and nodular dermatofibrosis) are advanced. Very promising are the results of gene therapy for certain canine monogenic diseases (haemophilia, hereditary retinal dystrophy, mucopolysaccharidosis), which have human equivalents. The above mentioned examples prove a very important model role of the dog in studies of human genetic diseases. On the other hand, the identification of gene mutations responsible for hereditary diseases has a substantial impact on breeding strategy in the dog. PMID- 15131352 TI - Milk trait heritability and correlation with heterozygosity in yak. AB - 382 yak cows were examined for milk yield, fat, protein and lactose contents. Six polymorphic loci, alphas1-CN, kappa-CN, beta-CN, beta-Lg, alpha-La and MUC-1, were scored by PAGE electrophoresis for each individual. The values of milk yield, fat, protein and lactose content were 247.13 kg, 5.81%, 5.18% and 4.93%, respectively. Based on the 6 polymorphism loci, the average heterozygosity of the yak population was 0.1794. Calculated by the marker-based method, heritability estimates for milk yield, fat, protein and lactose contents were 0.353 +/- 0.093, 0.316 +/- 0.101, 0.415 +/- 0.098 and 0.481 +/- 0.035, respectively. The relatively high or medium heritability of these traits indicate that it is feasible to rely directly on them in breeding for the improvement in a relatively short period. The significant linear regression between heterozygosity and fat percentage with a positive slope (R = 0.0420) indicated that inbreeding affected milk fat content in this population. PMID- 15131353 TI - Polymorphism within the bovine estrogen receptor-alpha gene 5'-region. AB - Due to the functions that estrogens play in the regulation of reproduction, development of the mammary gland, growth and differentiation of cells, estrogen receptors and their genes are considered candidates for the markers of production and functional traits in farm animals, including cattle. In the present study, on the basis of the sequences of the human, ovine, and porcine ER genes, available in the GenBank database, sets of PCR primers were designed and used to amplify the bovine ERalpha gene 5'-region. Seven overlapping fragments of the 5' region of the bovine ERalpha gene were amplified and then sequenced. Altogether, these fragments were composed in the 2853-bp sequence which was deposited in the GenBank database under accession no. AY340597. The sequenced fragment included the noncoding exons A, B, C, their putative promoters, and a part of the coding exon 1. A polymorphism within the 5' region of the bovine ERalpha gene-A/G transition, which could be recognized with RFLP-BglI, lying upstream to the exon C, was identified for the first time using this sequence. PMID- 15131354 TI - Role of epigenetic DNA alterations in the pathogenesis of systemic lupus erythematosus. AB - Epigenetic alternations in genomic DNA encompass cytosine methylation in cytosine and guanine (CpG) dinucleotide islands, which are usually extended in the promoter and first exon of genes. The DNA methylation is carried out by DNA methyltransferases (DNMT) and it serves as an epigenetic method of gene expression modulation. The epigenetic alternations in genomic DNA have been implicated in the development of malignant and autoimmune diseases. The epigenetic aberration in regulatory DNA sequences may also be responsible for the emergence of changes in the immune system in patients with systemic lupus erythematosus (SLE). The agents 5-azacytidine (azacitidine) and 5-aza-2' deoxycytidine (decitabine) belong to inhibitors of methyltransferase. These compounds affect the methylation level of promoter sequences and cause phenotypic changes in peripheral blood mononuclear cells (PBMC), which are similar to those observed in PBMC of SLE patients. The lack of methylcytosine in CpG dinucleotides may be responsible for the antigenic properties of microbial DNA. The presence of low-apoptotic methylated DNA fragments has been identified in plasma of SLE patients. These DNA fragments exhibit antigenic properties and may elicit the humoral response responsible for the flare of SLE. The low methylation of CpG residues in the regulatory sequences may also contribute to the elevated expression of human endogenous retroviruses (HERVs) in PBMC of SLE patients. The HERV components exhibit a profound similarity with nuclear antigens and may be responsible for the enhancement of the production of anti-antinuclear antibodies (ANA). Recent advances in the investigation of epigenetic DNA changes have formed the basis of improved understanding of etiopathogenesis of SLE, which may thereby facilitate improvement in therapeutic principles of this disease. PMID- 15131355 TI - Screening for mutations in the GJB3 gene in Brazilian patients with nonsyndromic deafness. AB - Deafness is a complex disorder that is affected by a high number of genes and environmental factors. Recently, enormous progress has been made in nonsyndromic deafness research, with the identification of 90 loci and 33 nuclear and 2 mitochondrial genes involved (http://dnalab-www.uia.ac.be/dnalab/hhh/). Mutations in the GJB3 gene, encoding the gap junction protein connexin 31 (Cx31), have been pathogenically linked to erythrokeratodermia variabilis and nonsyndromic autosomal recessive or dominant hereditary hearing impairment. To determine the contribution of the GJB3 gene to sporadic deafness, we analysed the GJB3 gene in 67 families with nonsyndromic hearing impairment. A single coding exon of the GJB3 gene was amplified from genomic DNA and then sequenced. Here we report on three amino acid changes: Y177D (c.529T > G), 49delK (c.1227C > T), and R32W (c.144-146delGAA). The latter substitution has been previously described, but its involvement in hearing impairment remains uncertain. We hypothesize that mutations in the GJB3 gene are an infrequent cause of nonsyndromic deafness. PMID- 15131356 TI - Ovarian cystadenoma as a characteristic feature of families with hereditary ovarian cancers unassociated with BRCA1 and BRCA2 mutations. AB - The study aimed to determine whether hereditary ovarian cancers that are not caused by BRCA1/BRCA2 constitutional mutations are associated with a predisposition to cystadenoma. The study consisted of two parts. Part one concerned the incidence of ovarian cystadenoma in females from families with hereditary ovarian cancer unassociated with BRCA1 mutations. The study group included 62 female patients from 29 families, without any previously diagnosed malignancy, with no proven constitutional mutation of the BRCA1 gene. The first control group was composed of 62 female patients from 53 families, without any previously diagnosed malignancy, with an identified constitutional mutation of the BRCA1 gene. The second control group comprised 124 female patients for whom the only reason for the examination was a prophylactic check-up. All studied women were subjected to intravaginal ultra- sonographic investigations. In 8 patients with benign and/or borderline ovarian cystadenoma, a complete sequencing of coding fragments of the BRCA2 gene from the peripheral blood DNA was performed. Part two of this study concerned the incidence and pattern of malignant tumors in the families of female patients with ovarian cystadenoma. The final study group included 117 patients who had 726 I0 relatives (359 females and 367 males). We concluded that cystadenoma is likely to be a characteristic feature of the subgroup of families with hereditary ovarian cancers unassociated with BRCA1/BRCA2 constitutional mutations. PMID- 15131357 TI - Chromosome aberrations, sister chromatid exchanges (SCE) and cell division kinetics in human lymphocytes exposed in vitro to purified trypsin inhibitor from Ascaris. AB - The purified trypsin inhibitor (TI) isolated from nematode Ascaris suum was tested in vitro for chromosome aberrations and sister chromatid exchanges (SCE). TI was obtained from the musculocutaneous sac homogenate of adult Ascaris by the modified method of Rola and Pudles. The inhibitor was isolated and purified from the SF5 fraction of proteins by gel filtration on Sephadex G-50 and electrophoresis SDS-PAGE of the obtained fraction after molecular filtration. TI showed a high inhibitory activity against crystalline trypsin (18.8 Kassell's units/mg of protein). Genotoxicity assessment of TI was carried out on metaphase plates received from peripheral blood lymphocyte macroculture (48 h-test of structural chromosome aberrations and 72 h-test of SCE), without exogenous metabolic activation. TI was tested in doses: 25, 50 and 100 microg per mL of culture. Kinetics of cell divisions was determined by the replication index (RI). We found that TI in vitro did not induce chromosome aberrations. It induced a higher number of SCE per cell but less than double frequency as compared to the control. The difference was significant only for the dose 50 microg/mL. For all doses, replication index (RI) values were significantly higher and mitotic index (MI) values were significantly lower than in the control. Thus the Ascaris trypsin inhibitor did not show any genotoxic properties but exhibited a mitostatic activity. PMID- 15131358 TI - Only neutral polymorphisms found in the TIGR/myocilin gene of 45 Polish patients with primary open-angle glaucoma. AB - The aim of the study was to identify mutations of the TIGR gene in Polish patients with primary open-angle glaucoma (POAG) and to define possible genotype phenotype correlations. The study included 45 patients with a verified diagnosis of POAG. The PCR amplification of all three exons of the TIGR gene and screening for the sequence changes by CSGE analysis was done for every patient. The probes with identified heteroduplexes were sequenced. Altogether 315 PCR products were obtained. The CSGE analysis detected 60 possible changes of the sequence in 28 patients. 34 heteroduplexes were chosen for sequencing, including 29 unique changes and 5 changes representative of identical heteroduplexes. Direct sequencing enabled detection of only four different changes in the TIGR gene sequence. Three of them: 5'UTR -83G-->A (in 14 patients), +227 exon 1 G-->A, Arg76Lys (in 14 patients) and +311 exon 3 T-->C, Tyr347Tyr (in 4 patients) have already been described in the literature as neutral polymorphisms of the gene. Only one change in the promoter, 5'UTR -126T-->C (in 2 patients), has not been described in the literature to date. However, this change does not alter directly the sequence of amino acids in myocilin, so it is difficult to conclude on its pathogenetic role. Thus our study showed only neutral polymorphisms of the TIGR gene. This suggests that the patients probably have mutations in other genes, so other loci that predispose to POAG must be analyzed. PMID- 15131359 TI - The alkali hypophosphites KH2PO2, RbH2PO2 and CsH2PO2. AB - The structures of the hypophosphites KH(2)PO(2) (potassium hypophosphite), RbH(2)PO(2) (rubidium hypophosphite) and CsH(2)PO(2) (caesium hypophosphite) have been determined by single-crystal X-ray diffraction. The structures consist of layers of alkali cations and hypophosphite anions, with the latter bridging four cations within the same layer. The Rb and Cs hypophosphites are isomorphous. PMID- 15131360 TI - Sodium magnesium bis(vanadate) pyrovanadate: Na6Mg2(VO4)2(V2O7). AB - The crystal structure of the new complex vanadium oxide Na(6)Mg(2)(VO(4))(2)(V(2)O(7)) was solved from X-ray single-crystal data. The structure contains VO(4) tetrahedra and MgO(6) octahedra, linked by corners and forming a complex three-dimensional framework. A half of the VO(4) tetrahedra are connected only to MgO(6) octahedra, whereas the others are corner-sharing, forming V(2)O(7) pyrovanadate groups with statistically random orientations. One unique Mg atom is located at an inversion centre, while the other Mg atom, one unique V atom and five unique O atoms lie on mirror planes. PMID- 15131361 TI - Oxygen-deficient strontium cobaltate, SrCoO2.64. AB - Single crystals of strontium cobaltate, SrCoO(3-x), have been grown by the floating-zone method in an oxygen flow. The compound crystallizes with the cubic perovskite structure, with Pm-3m symmetry, as determined by X-ray diffraction. Refinement of the O-atom site occupancy yields the chemical composition SrCoO(2.64) [x = 0.36 (3)]. The anisotropic displacement ellipsoids of the O atoms suggest that their positional disorder occurs in a direction perpendicular to that of the Co-O bonds. PMID- 15131362 TI - catena-Poly[europium(III)-tri-micro-2,3-dimethoxybenzoato]. AB - The title compound, [Eu(C(9)H(9)O(4))(3)](n) or [Eu(2,3-DMOBA)(3)](n), where 2,3 DMOBA is 2,3-dimethoxybenzoate, is an infinite one-dimensional non centrosymmetric coordination polymer. The unique Eu(III) atom is bridged by six carboxylate ligands; it is ennea-coordinated and has a distorted tricapped trigonal prism geometry. The Eu-O distances are in the range 2.315 (3)-2.959 (5) A. PMID- 15131363 TI - Rubidium bis(trifluoromethanesulfonyl)imidate dioxane disolvate. AB - The bis(trifluoromethanesulfonyl)imidate anion crystallizes with Rb as the title dioxane 1:2 solvate, Rb(+).CF(3)SO(2)NSO(2)CF(3)(-).2C(4)H(8)O(2), with the anion in a transoid conformation, as opposed to the cisoid form typically seen when there are significant cation-anion interactions. The Rb(I) cation is eight coordinate, interacting with one anion in a chelating fashion and with two other anions through the remaining sulfonyl O atoms. The latter interactions link ion pairs through the formation of Rb(2)O(2) dimers about inversion centers at (0, 1/2, 0) and (1/2, 1/2, 0), forming extended columns which run parallel to the a axis of the unit cell. Rb-dioxane bridges crosslink these salt columns in the (010), (001) and (011) directions, resulting in a three-dimensional network solid. One dioxane solvent molecule is disordered over two half-occupancy sites. PMID- 15131364 TI - 1,4-Diazabicyclo[2.2.2]octane-1,4-diium trichromate. AB - The title compound, (C(6)H(14)N(2))[Cr(3)O(10)], consists of a diazabicyclo[2.2.2]octane-1,4-diium cation and a discrete trichromate anion linked by an N-H.O hydrogen bond. Three CrO(4) tetrahedra are joined via shared O atoms to form the trichromate anion. Supramolecular rings, which can be described by the graph-set motif R(4)(4)(26), are built via N-H.O hydrogen bonds, and C-H.O interactions play lesser roles in forming the structure. PMID- 15131365 TI - Chloro(histamine)(1,10-phenanthroline)copper(II) chloride monohydrate. AB - In the cationic complex present in the title compound, chloro[2-(4-imidazolyl kappaN(1))ethylamine-kappaN](1,10-phenanthroline-kappa(2)N,N')copper(II) chloride monohydrate, [CuCl(C(5)H(9)N(3))(C(12)H(8)N(2))]Cl.H(2)O, the metal centre adopts a five-coordinate geometry, ligated by the two phenanthroline N atoms, two amine N atoms of the histamine ligand (one aliphatic and one from the imidazole ring) and a chloro ligand. The geometry around the Cu atom is a distorted compressed trigonal bipyramid, with one phenanthroline N and one imidazole N atom in the axial positions, and the other phenanthroline N atom, the histamine amine N atom and the chloro ligand in the equatorial positions. The structure includes an uncoordinated water molecule, and a Cl(-) ion to complete the charge. The water molecule is hydrogen bonded to both Cl(-) ions (coordinated and uncoordinated), and exhibits a close Cu.H contact in the equatorial plane of the bipyramid. PMID- 15131366 TI - A metal-mediated dimerization of the ligand bis(N,N diethylamino)carbeniumdithiocarboxylate. AB - The title compound, methylene bis[bis(N,N diethylamino)carbeniumdithiocarboxylate] pentachlorooxorhenium, (C(21)H(42)N(4)S(4))[ReCl(5)O], is the result of an unusual dimerization of the ligand bis(N,N-diethylamino)carbeniumdithiocarboxylate [(Et(2)N)(2)C(2)S(2)] upon reaction with [ReOCl(3)(PPh(3))(2)] in chloroform under reflux conditions. The compound was obtained as a dicationic moiety, with the molecular [ReOCl(5)](2-) anion providing the charge compensation. The planes of the carbenium and thiocarboxylate moieties are nearly perpendicular to one another and the backbone C-C bond length in the N(2)CCS(2) group is the same as a normal C-C single-bond length. PMID- 15131367 TI - Tris[N-(4-fluorophenyl)pyridine-2-carboxamidato-kappa2N,N']cobalt(III) trihydrate: a novel meridional complex. AB - The title complex, [Co(C(12)H(8)FN(2)O)(3)].3H(2)O, has been synthesized for the first time. The complex comprises three bidentate ligands containing the pyridine 2-carboxamide stem. The distorted octahedral coordination around the Co atom is formed via the pyridine (py) N atom and the deprotonated amide N atom of each ligand, with the three pyridine rings in a meridional arrangement. For each ligand, the pyridine ring and the carbonyl group are nearly coplanar, with torsion angles in the range 0.4 (3)-4.8 (4) degrees. The Co-N(py) distances [1.9258 (16)-1.9656 (17) A] are shorter than the corresponding Co-N(amide) distances [1.9372 (17)-1.9873 (15) A]. In addition, the Co-N(py) distances are closely related to the magnitudes of the chelate angles, a shorter Co-N(py) distance corresponding to a larger angle. Five intermolecular hydrogen bonds, involving carbonyl O atoms of the ligands and lattice water molecules, lead to the formation of a mesh structure. PMID- 15131368 TI - A heterometallic polymeric complex: [Cu2(N3)2(medpt)2[Ni(CN)4]]n [medptis bis(3 aminopropyl)methylamine]. AB - The structure of the title compound, catena-poly[[di-micro-azido kappa(4)N(1):N(1)-bis[[bis(3-aminopropyl)methylamine-kappa(3)N]copper(II)]]-micro cyano-[dicyanonickel(II)]-micro-cyano], [Cu(2)(N(3))(2)(medpt)(2)[Ni(CN)(4)]](n) [medpt is bis(3-aminopropyl)methylamine, C(7)H(19)N(2)] or [Cu(2)Ni(CN)(4)(N(3))(2)(C(7)H(19)N(3))(2)](n), is a one-dimensional heterometallic covalent chain where Ni(CN)(4)(2-) functions as a molecular ion bridge. The Ni atom sits on the centre of inversion. The chain undergoes hydrogen bonding interactions, forming a three-dimensional supramolecular network. PMID- 15131369 TI - The catena-arsenite chain anion, [AsO2](n)(n-): (H3NCH2CH2NH3)0.5[AsO2] and NaAsO2 (revisited). AB - The title compounds contain the catena-arsenite [AsO(2)](n)(n-) unit, in which the As(III) atom is pyramidally coordinated to one terminal and two bridging O atoms, resulting in an infinite anionic chain. Ethylenediammonium catena arsenite, (C(2)H(10)N(2))(0.5)[AsO(2)], is the first example of this anion in the company of an organic cation. The ethylenediammonium species interact with the [AsO(2)](-) chains by way of N-H.O hydrogen bonds. The structure of sodium catena arsenite, Na[AsO(2)] [Menary (1958). Acta Cryst. 11, 742-743], has been redetermined to yield more reliable geometrical parameters. The As-O distances are normal and the Na(+) cation is seven-coordinate [Na-O = 2.285 (4)-3.063 (4) A] in a distorted capped trigonal prismatic geometry. PMID- 15131370 TI - Bis[1,1-dimethylbiguanide(1-)-kappa2N2,N5]copper(II) monohydrate. AB - The crystal structure of the title compound, [Cu(C(4)H(10)N(5))(2)].H(2)O, contains two independent copper N,N-dimethylbiguanide complex units, each with square-planar coordination of the Cu atom by four N atoms. The two complexes have different symmetry, with one Cu atom lying on an inversion centre and the other on a twofold rotation axis. The Cu-N bond lengths are 1.923 (2) and 1.950 (2) A in the centrosymmetric complex, and 1.928 (2) and 1.938 (2) A in the non centrosymmetric complex. The crystal structure is stabilized by N-H...O, O-H...N and N-H...N hydrogen bonds; each water molecule forms four hydrogen bonds involving three different Cu complexes. PMID- 15131371 TI - A novel bridged asymmetric binuclear manganese(II) complex with DTPB [DTPB is 1,1,4,7,7-pentakis(1H-benzimidazol-2-ylmethyl)-1,4,7-triazaheptane]. AB - The crystal structure of the title compound, tetrachloro[micro-1,1,4,7,7 pentakis(1H-benzimidazol-2-ylmethyl)-1,4,7-triazaheptane]dimanganese(II) methanol pentasolvate tetrahydrate, [Mn(2)Cl(4)(C(44)H(43)N(13))].5CH(4)O.4H(2)O, contains an asymmetric dinuclear Mn(II)-DTPB [DTPB is 1,1,4,7,7-pentakis(1H-benzimidazol-2 ylmethyl)-1,4,7-triazaheptane] complex with an intra-ligand bridging group ( NCH(2)CH(2)N-), as well as several solvate molecules (methanol and water). Both Mn(II) cations have similar distorted octahedral coordination geometries. One Mn(II) cation is coordinated by a Cl(-) anion and five N atoms from the ligand, and the other is coordinated by three Cl(-) anions and three N atoms of the same ligand. The Mn.Mn distance is 7.94 A. A Cl.H-O.H-O.H-N hydrogen-bond chain is also observed, connecting the two parts of the complex. PMID- 15131372 TI - Bis(pentane-1,5-diammonium) decaiodotriplumbate(II). AB - The title compound, [(NH(3)C(5)H(10)NH(3))(2)[Pb(3)I(10)]](n), crystallizes as an organic-inorganic hybrid. As such, the structure consists of extended chains of [Pb(3)I(10)](n)(4n-) ions extending along [111]. The asymmetric unit contains two independent Pb atoms: one is in a general position and the other is on an inversion centre. Each Pb atom is octahedrally coordinated by six iodide ions and exhibits both face- and edge-sharing with adjacent atoms in the inorganic chain. The organic counter-ion, viz. pentane-1,5-diammonium, lies in channels formed by the chains and interacts with these chains via N-H.I hydrogen bonding. PMID- 15131373 TI - Two furopyridine complexes of nickel(II) isothiocyanate. AB - Pyridine fused with a furan ring (fupy), and its dimethyl derivative, have been used for the first time as ligands to synthesize potentially new Werner clathrates. The extended aromatic system of pyridine-like ligands influences considerably the molecular structure of prepared nickel complexes. The molecular structure of tetrakis(furo[3,2-c]pyridine)bis(isothiocyanato)nickel(II) tetrahydrofuran (THF) solvate, [Ni(NCS)(2)(C(7)H(5)NO)(4)].C(4)H(8)O or [Ni(NCS)(2)(fupy)(4)].THF, (I), reveals a 'four-blade propeller' arrangement of ligands, with the angles between the fupy planes and the basal octahedron plane spanning the range 38.7-55.3 degrees. These angles are much larger (69.9-78.8 degrees ) in the centrosymmetric complex tetrakis(2,3-dimethylfuro[3,2 c]pyridine)bis(isothiocyanato)nickel(II) 6.6-hydrate, [Ni(NCS)(2)(C(9)H(9)NO)(4)].6.6H(2)O or [Ni(NCS)(2)(Me(2)fupy)(4)].6.6H(2)O, (II), in which crystallographically imposed inversion symmetry is present. PMID- 15131374 TI - 3-(4-Acetylphenyl)-1-(4-nitrophenyl)triazene. AB - The crystal structure of the title compound, C(14)H(12)N(4)O(3), shows that the stereochemistry about the N=N double bond of the N=N-N(H) moiety is trans. The whole molecule is almost planar (r.m.s. deviation = 0.0654 A), the interplanar angle between the phenyl rings being 0.7 (1) degrees and the largest interplanar angle being that between the phenyl ring and the nitro group of the 4-nitrophenyl substituent [11.5 (2) degrees ]. Intermolecular N-H.O interactions between molecules related by translation give rise to chains along the [110] and [1-10] directions, and these chains are held together by N.O pi-pi interactions. An unequal distribution of the double-bond character among the N atoms suggests a delocalization of pi electrons over the diazoamine group and the adjacent aryl substituents. PMID- 15131375 TI - 3,3,4,4,5,5-Hexafluoro-1,2-bis(5-hydroxymethyl-2-methyl-3-thienyl)cyclopent-1 ene. AB - The title compound, C(17)H(14)F(6)O(2)S(2), a photochromic diarylethene, is one of the most promising materials for optical memories and other optoelectronic devices. The hexafluorocyclopentene group and the two thiophene rings are all planar, and the dihedral angles between the cyclopentene ring and the adjacent thiophene rings are 46.4 (1) and 49.5 (1) degrees. PMID- 15131376 TI - [(E)- and [(Z)-2-[alpha,beta-bis(methoxycarbonyl)vinyl]cyclopentadien-1 ylidene]triphenylphosphorane. AB - The Ramirez ylide undergoes electrophilic substitution with dialkyl acetylenedicarboxylates, yielding a mixture of the Z and E adducts. The crystal structure analyses of the two adducts formed using dimethylacetylene, viz. dimethyl (E)- and (Z)-1-[2-(triphenylphosphoranylidene)cyclopentadien-1 yl]ethylenedicarboxylate, both C(29)H(25)O(4)P, explain an unusual chemical shift observed for the vinyl H atom of the Z adduct, which had previously precluded a definitive assignment of the isomers. In addition, the structures explain why only one of the isomers reacts further with acetylene esters to produce azulenes with a rare substitution pattern. PMID- 15131377 TI - Methyl 2,6-di-O-benzyl-3,4-O-phenylphosphinediyl-beta-D-galactopyranoside(P B)borane with chirality on the P atom. AB - A carbohydrate-derived optically active P-chiral dioxophenylphospholane-borane complex, C(27)H(32)BO(6)P, was prepared from bis(diethylamino)phenylphosphine and methyl 2,6-di-O-benzyl-beta-D-galactopyranoside. The phosphinite was prepared with high diastereoselectivity and in good yield. The absolute configuration (R) at the P atom was deduced from the known configuration of the sugar moiety. Weak intermolecular interactions link the molecules into a three-dimensional network. PMID- 15131378 TI - 5,5-Dimethoxy-2-phenoxy-1,3,2-dioxaphosphorinane 2-oxide. AB - In the title compound, C(11)H(15)O(6)P, the six-membered dioxaphosphorinane ring of the cyclic phosphate triester exists in a distorted chair conformation, with the phenoxy group in an axial position. The phenyl ring and both methoxy groups are in a trans-gauche orientation with respect to the 1,3,2-dioxaphosphorinane ring. In the phosphate group, a significant deformation from the ideal tetrahedral shape is observed. The crystal structure is stabilized by a three dimensional network of C-H...O interactions. PMID- 15131379 TI - 5-(2-Chlorophenyldiazenyl)salicylaldehyde and 4-(2-chlorophenyldiazenyl)-2 [[tris(hydroxymethyl)methyl]aminomethylene]cyclohexa-3,5-dien-1(2H)-one. AB - The molecule of the former title compound, C(13)H(9)ClN(2)O(2), (I), is nearly planar, with an intramolecular O...O hydrogen bond of 2.692 (2) A. The latter title compound, C(17)H(18)ClN(3)O(4), (II), exists in the keto-amine tautomeric form, with a strong intramolecular hydrogen bond of 2.640 (2) A between the O and N atoms, the H atom being bonded to the N atom. The azobenzene moieties of both molecules have trans configurations, and the dihedral angle between the planes of the two aromatic rings is 4.1 (1) degrees in (I) and 9.9 (1) degrees in (II). The N-H...O hydrogen-bonded rings are almost planar and coupled with the cyclohexadiene rings in (II). PMID- 15131380 TI - Two oxazane macrocycles. AB - The 20-membered ring in 1,7,11,17-tetraoxa-2,6,12,16-tetraazacycloeicosane tetrahydrochloride, C(12)H(32)N(4)O(4)(4+).4Cl(-), adopts an endo conformation, while the 18-membered ring in 1,6,10,15-tetraoxa-2,5,11,14 tetraazacyclooctadecane tetrahydrochloride, C(10)H(28)N(4)O(4)(4+).4Cl(-), lies about an inversion centre and adopts a symmetrical conformation. In the crystal structures of both compounds, the cations and chloride anions are linked by N H...Cl hydrogen bonds into planar sheets of molecules; the sheets are linked into three-dimensional networks via C-H...Cl hydrogen bonds. PMID- 15131381 TI - N,N'-Propylenedioxybis(2,4,6-trimethylbenzenesulfonamide): molecules of unexpected conformation form a molecular ladder built from two independent N H...O=S hydrogen bonds. AB - Molecules of the title compound, C(21)H(30)N(2)O(6)S(2), adopt a skeletal conformation which does not possess even approximate internal symmetry. The molecules are linked by two N-H...O=S hydrogen bonds [H...O = 1.97 A (x2), N...O = 2.865 (2) and 2.864 (2) A, and N-H...O = 160 and 159 degrees] into molecular ladders, alternatively described as chains of edge-fused R(2)(2)(20) rings. PMID- 15131382 TI - Hydrogen bonding in substituted nitroanilines: hydrogen-bonded sheets in 4-iodo-3 nitroaniline. AB - In the title compound, C(6)H(5)IN(2)O(2), the nitro group is disordered over two sets of sites, each with 0.5 occupancy, and the amino N atom is pyramidal. The molecules are linked into sheets by a combination of three-centre N-H...(O)(2) hydrogen bonds involving alternative pairs of O-atom sites and two-centre N-H...N hydrogen bonds involving the pyramidal amino group. PMID- 15131383 TI - Triphenyl(3,4,5-triiodophenyl)methane. AB - The crystal structure of the title compound, alternatively called 1,2,3-triiodo-5 (triphenylmethyl)benzene, C(25)H(17)I(3), is analysed in terms of I.I and I.pi interactions and the herring-bone T motif between phenyl groups. There are two molecules in the asymmetric unit, denoted A and B. Inversion-related A molecules are connected via an I.pi interaction (3.641 A, to a C-C bond mid-point) to form an I.pi dimer, and these dimers are connected through symmetry-independent B molecules via I.I [3.5571 (15) A] and I.pi (3.561 A, to a C-C bond mid-point) interactions. PMID- 15131384 TI - 2-Amino-4-(1-naphthyl)-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3-carbonitrile and 2 amino-7,7-dimethyl-4-(1-naphthyl)-5-oxo-5,6,7,8-tetrahydro-4H-chromene-3 carbonitrile. AB - Syntheses and X-ray structural investigations have been carried out for the two title compounds, C(20)H(16)N(2)O(2), (IIIa), and C(22)H(20)N(2)O(2), (IIIb). In (IIIa), the heterocyclic ring adopts a sofa conformation, while in (IIIb), the ring has a flattened boat conformation. In both molecules, the fused cyclohexenone ring adopts a sofa conformation. The dihedral angles between these two flat fragments are 3.5 (2) and 17.5 (2) degrees in (IIIa) and (IIIb), respectively. The dihedral angles between the pseudo-axial naphthalene substituents and the planes of the pyran rings are 90.9 (1) and 96.7 (1) degrees, respectively. In the crystal structure of (IIIa), intermolecular N-H.N and N H...O hydrogen bonds link the molecules into infinite tapes along the b axis, while molecules of (IIIb) form centrosymmetric dimers via N-H...N hydrogen bonds, with only one H atom of the NH(2) donor group taking part in hydrogen bonding. PMID- 15131385 TI - A substituted EDOT precursor: diethyl 3,4-dihydroxythiophene-2,5-dicarboxylate. AB - The title compound, C(10)H(12)O(6)S, has been obtained as dark-yellow chunk shaped crystals, together with the expected thin white needles. The structures of the two phases are identical. Two independent molecules compose the asymmetric unit: one molecule is totally planar, whereas a methyl group of the second molecule points out of the plane. Each molecule participates in several intra- and intermolecular hydrogen bonds and short contacts. The overall structure can be regarded as parallel sheets of molecules. Within a sheet, molecules are connected to one another in an infinite network via numerous short intermolecular contacts. Sheets are connected via hydrogen bonds and short contacts, in particular involving the methyl groups. PMID- 15131386 TI - 1-(4-Bromophenyl)-2-methyl-4-nitro-1H-imidazole: bifurcated Br...O halogen-nitro bonds. AB - The crystal packing of the title compound, C(10)H(8)BrN(3)O(2), is determined mainly by relatively strong bifurcated C-Br...O halogen-nitro bonds. Both O atoms are involved in this interaction in an almost symmetrical manner and the difference [0.078 (3) A] between the Br...O contact lengths is one of the smallest found in similar compounds. Halogen bonds and weak hydrogen bonds connect molecules into layers which are stacked along the [100] direction. PMID- 15131387 TI - 3-formylphenylboronic acid. AB - The molecule of the title compound, C(7)H(7)BO(3), is planar, and the bond lengths and angles are typical. The formyl group is essentially coplanar with the benzene ring but does not influence significantly the distortion of the ring, although the formyl group does have a strong influence on the crystal packing. The geometry of the boronic acid group is typical. In the crystal structure, the molecules are linked by O-H...O hydrogen bonds. PMID- 15131388 TI - (R)-2,3,4,5-Tetrahydro-2,2,4-trimethyl-1,5-benzothiazepine and rac-5-benzoyl 2,3,4,5-tetrahydro-2,2,4-trimethyl-1,5-benzothiazepine: chains built from N-H... Sand C-H...pi(arene) hydrogen bonds. AB - Molecules of (R)-2,3,4,5-tetrahydro-2,2,4-trimethyl-1,5-benzothiazepine, C(12)H(17)NS, are linked into spiral C(5) chains by a single N-H...S hydrogen bond, while molecules of rac-5-benzoyl-2,3,4,5-tetrahydro-2,2,4-trimethyl-1,5 benzothiazepine, C(19)H(21)NOS, are linked into zigzag chains by a C H...pi(arene) hydrogen bond. PMID- 15131389 TI - 2,3-Bis[(4-methylphenyl)sulfanyl]-4H-1-benzothiopyran-4-one 1,1-dioxide forms a framework built from C-H...O, C-H...pi(arene) and pi-pi interactions. AB - Molecules of the title comound, C(23)H(18)O(3)S(3), are linked into sheets by a combination of one C-H...O hydrogen bond and two C-H...pi(arene) hydrogen bonds, and these sheets are weakly linked into a three-dimensional structure by means of a single aromatic pi-pi stacking interaction. PMID- 15131390 TI - 1-Bromo-2,6-dihydroxybenzene containing R(4)(4)(8) rings and C(2) helices. AB - Molecules of the title compound (also known as 2-bromoresorcinol), C(6)H(5)BrO(2), are essentially planar and possess normal geometrical parameters. The crystal packing is influenced by O-H...O and O-H...O/Br hydrogen bonds and pi pi stacking interactions, resulting in a distinctive high-symmetry structure containing R(4)(4)(8) rings and helical C(2) chains. PMID- 15131391 TI - 6-(2-Fluorobenzylidene)-2-[1-(2-fluoro-4-biphenyl)ethyl]thiazolo[3,2 b][1,2,4]triazol-5(6H)-one. AB - The title compound, C(25)H(17)F(2)N(3)OS, was synthesized from 6 (benzylidene)thiazolo[3,2-b][1,2,4]triazol-5(6H)-one. The fused thiazolo[3,2 b][1,2,4]triazole system is essentially planar, and bifurcated C-H.O, C-H.N and C H.F interactions are present between molecules. PMID- 15131392 TI - N,N'-Bis(dipiperidin-1-ylmethylene)propane-1,3-diamine and N,N'-bis(1,3 dimethylperhydropyrimidin-2-ylidene)propane-1,3-diamine. AB - The molecular structures of the title compounds, C(25)H(46)N(6) and C(15)H(30)N(6), respectively, show the two guanidyl moieties each connected by propyl bridges. The different substitution pattern of the guanidyl groups has no influence on the distinct localization of their C=N double bonds. Both compounds exhibit approximate twofold symmetry and the crystal packing shows no prominent hydrogen-bonding interactions. PMID- 15131393 TI - 2-(4-Nitrophenoxy)benzoic acid: a three-dimensional hydrogen-bonded framework in a triclinic structure having Z' = 3. AB - The title compound, C(13)H(9)NO(5), crystallizes in space group P-1, with Z' = 3. The molecules are linked by O-H...O hydrogen bonds [H...O = 1.79-1.81 A, O...O = 2.625 (3)-2.648 (3) A and O-H...O = 172-176 degrees ] into two types of R(2)(2)(8) dimer, only one of which is centrosymmetric. An extensive series of soft hydrogen bonds, of C-H...O and C-H...pi(arene) types, links the dimers into a three-dimensional framework. PMID- 15131394 TI - Monoclinic pseudosymmetry in 2-phenoxybenzenesulfonamide, a triclinic structure having Z' = 4, and spontaneous resolution in monoclinic N-methyl-2 phenoxybenzenesulfonamide. AB - 2-Phenoxybenzenesulfonamide, C(12)H(11)NO(3)S, (I), crystallizes in space group P 1 with Z' = 4, but the structure closely mimics the monoclinic space group P2(1)/b with Z' = 2. The molecules of (I) are linked by a combination of N-H...O and C-H...O hydrogen bonds into two independent chains of centrosymmetric edge fused R(2)(2)(18) and R(6)(6)(34) rings. N-Methyl-2-phenoxybenzenesulfonamide, C(13)H(13)NO(3)S, (II), crystallizes in space group P2(1) with Z' = 1, and is an example of spontaneous resolution. The molecules are linked by N-H...O and C H...O hydrogen bonds into chains of spiro-fused R(2)(2)(12) rings, and these chains are linked into sheets by a single C-H...pi(arene) hydrogen bond. PMID- 15131395 TI - trans-4-Bromo-ONN-azoxybenzene at 100 K. AB - The crystal structure of the alpha isomer of trans-4-bromoazoxybenzene [systematic name: trans-1-(bromophenyl)-2-phenyldiazene 2-oxide], C(12)H(9)BrN(2)O, has been determined by X-ray diffraction. The geometries of the two molecules in the asymmetric unit are slightly different and are within approximately 0.02 A for bond lengths, approximately 2 degrees for angles and approximately 3 degrees for torsion angles. The azoxy bridges in both molecules have the typical geometry observed for trans-azoxybenzenes. The crystal network contains two types of planar molecules arranged in columns. The torsion angles along the Ar-N bonds are only 7 (2) degrees, on either side of the azoxy group. PMID- 15131396 TI - L-Isoleucyl-L-phenylalanine dihydrate. AB - The structure of the title compound, C(15)H(22)N(2)O(3).2H(2)O, was derived from data collected on a very thin twinned needle. The peptide molecule is in a rare conformation normally associated with hydrophobic dipeptides that form nanotubes. Nevertheless, the present structure is divided into hydrophobic and hydrophilic layers. PMID- 15131397 TI - Two stereoisomers of the rat toxicant norbormide. AB - The structures of two diastereoisomers of norbormide [systematic name: 5 [hydroxy(phenyl)(2-pyridyl)methyl]-8-[phenyl(2-pyridyl)methylene]-3a,4,7,7a tetrahydro-4,7-methano-1H-isoindole-1,3(2H)-dione], viz. the unsolvated molecule, C(33)H(25)N(3)O(3), and the ethyl acetate hemisolvate, C(33)H(25)N(3)O(3).0.5C(4)H(8)O(2), have been determined unambiguously. They differ in the relative stereochemistry about the exocyclic double bond and the relative conformations of the aryl rings. Each compound exhibits both intra- and intermolecular hydrogen bonding. PMID- 15131398 TI - Vindoline and 16-demethoxyvindoline: two catharanthus-derived alkaloids. AB - Vindoline, C(25)H(32)N(2)O(6), and 16-demethoxyvindoline, C(24)H(30)N(2)O(5), both of which are naturally occurring biologically active products derived from plants, are important as possible starting materials for the synthesis of valuable anticancer antibiotics, viz. vincristine and vinblastine, and other pharmaceuticals. The vindoline framework consists of two five- and three six membered condensed rings. One of the six-membered rings adopts a boat conformation, one adopts a sofa conformation and the third is planar. Both five membered rings have envelope structures. The intramolecular hydrogen bonds present in the structures are characteristic of vinca alkaloids. PMID- 15131399 TI - Cancer variation associated with the position of the mutation in the BRCA2 gene. AB - Inherited mutations of the BRCA2 gene give rise to a multi-site cancer phenotype which includes breast cancer (in female and males), ovarian, pancreatic and prostate cancer, ocular and other melanomas, laryngeal, colon and stomach cancers. Interpretation of test results and risk assessment is therefore complex. It has been proposed that families with mutations in the ovarian cancer cluster region (OCCR) of exon 11 (nucleotides 3035-6629) express a higher ratio of ovarian to breast cancer, than families with mutations elsewhere in the BRCA2 gene. In this study we have investigated the presence of 7 types of cancer (ovary, male breast, pancreas, prostate, colon, stomach and melanoma) in first- and second-degree relatives of mutation-positive individuals in 440 families with a BRCA2 mutation. We reviewed histories of cancer in relatives among families with mutations distributed throughout the gene. Families with ovarian cancer were more likely to harbour mutations in the OCCR (nucleotides 3035-6629) than elsewhere in the gene (OR = 2.21; P = 0.0002). We also compared cancer risks according to ethnic group. Ashkenazi Jewish families with the 6174delT founder mutation were more likely to have a family member with ovarian cancer (OR = 1.58; P = 0.002) and less likely to have a family member with prostate cancer (OR = 0.62; P = 0.04) than were non-Jewish families. In contrast, a reduced presence of ovarian cancer was found in families of French-Canadian ancestry, compared to other ancestries (OR = 0.37; P = 0.0026). A high risk of male breast cancer was observed with the 6503delTT mutation (OR = 15.7; P = 0.023). Families of Polish ancestry had a reduced frequency of pancreatic cancer (OR = 0.0; P = 0.03) compared to families of other ethnic origins. In conclusion, both the position of mutation and the ethnic background of the family appear to contribute to the phenotypic variation observed in families with BRCA2 mutations. PMID- 15131400 TI - The 1100delAT BRCA1 and the 8765delAG BRCA2 mutations: occurrence in high-risk non-Ashkenazi Jews and haplotype comparison of Jewish and non-Jewish carriers. AB - Few mutations have been described in BRCA1 and BRCA2 in high-risk non-Ashkenazi Jews. In a Libyan family the 1100delAT BRCA1 mutation was detected and the 8765delAG BRCA2 mutation was previously described in two Jewish-Yemenite families. In this study, the rate of these mutations in high-risk Jews of North African and Yemenite origin was assessed, and the BRCA1 -linked haplotype of Jewish and non-Jewish 1100delAT mutation carriers were compared. Genotyping included 64 high-risk Yemenite women (tested only for the BRCA 2 mutation) and 147 high-risk North African women, tested for both mutations. PCR amplification was followed by either restriction enzyme digestion or DGGE or dHPLC analyses and direct sequencing. For haplotyping, 5 BRCA1 -linked markers were used. Neither the 1100delAT BRCA1 nor the 8765delAG BRCA2 mutations were detected in any non Ashkenazi individual. The haplotype of the non-Jewish 1100delAG mutation carrier differed from that of the Jewish-Libyan mutation carriers. We conclude that both1100delAT BRCA1 and 8765delAG BRCA2 mutations occur rarely in high-risk non Ashkenazi Jews, and while the latter seems to be a founder mutation in some populations, the former occurs on a different background in ethnically diverse families. PMID- 15131401 TI - BRCA1 testing in breast and/or ovarian cancer families from northeastern France identifies two common mutations with a founder effect. AB - OBJECTIVE: The purpose of this study was to determine whether two mutations detected frequently in a population of breast and/or ovarian cancer families originating from the northeastern part of France could be due to a founder effect. METHODS: 83 index cases of families ascertained to have a familial breast and/or ovarian cancer history, were screened for mutations in all coding exons of the BRCA1 gene, using combined DGGE and direct sequencing. For haplotype analysis, six polymorphic markers were used for allelotyping of mutation carriers and non carriers from nine families with 3600del11 mutation and four families with G1710X mutation. RESULTS: Of 83 index cases, 27 (32%) had 14 different BRCA1 mutations, one of which (G1710X), had not been reported in other populations. Two mutations were particularly common: 3600del11 in exon 11 accounted for 37% and the nonsense mutation G1710X in exon 18 for 15% of all mutations. We identified a common haplotype for each mutation suggesting a common founder for each recurrent mutation. No specific phenotype could be assigned to any of the common mutations. CONCLUSIONS: These data demonstrate geographical clustering and suggest a founder effect for particular BRCA1 mutations, which identification will facilitate carrier detection in French families with breast cancer and breast and/or ovarian cancer. PMID- 15131402 TI - BRCA1 testing with definitive results: a prospective study of psychological distress in a large clinic-based sample. AB - AIM: To examine the short-term psychological impact of receiving definite results concerning BRCA1 mutation status in a clinical setting. METHODS: A test was offered for consecutive sample of 395 women from 53 families with demonstrated BRCA1 mutations. The sample included 50 women with a personal history of cancer, and 345 women without. Of the 287 women who chose to be tested and participated in the study, 79% of those with cancer 33% of those without cancer had a demonstrated BRCA1 mutation. Psychological distress was measured with the hospital anxiety and depression scale (HADS), the general health questionnaire (GHQ-28), the impact of event scale (IES) and Beck's hopelessness scale (BHS) at the time the patients were offered testing and six weeks after receiving the test result. RESULTS: No significant changes were found in psychological distress from baseline to follow-up in any groups. Women with cancer were significantly more distressed than those without both at baseline and at six weeks, while women without cancer had levels of psychological distress comparable to or lower than normative data as measured by HADS. CONCLUSIONS: Our sample had a low level of psychological distress at baseline. Receiving a definite positive or negative result on the BRCA1 test had minimal effects on short-term psychological distress. These findings indicate that establishing a special psychological service in relation to predictive BRCA1 testing could be superfluous. PMID- 15131403 TI - An audit of screening for familial breast cancer before 50 years in the South Thames Region - have we got it right? AB - We have carried out an audit of breast screening by mammography under 50 years of age in a cohort of 192 women attending family cancer clinics run by the South Thames genetic services. Of these women, six came from families in which a BRCA mutation had been identified, 61 had > 50%, 35 a 20-50% and 90 had < 20% chance of carrying a high risk mutation. In the 192 women in the screened cohort, 9 breast cancers were diagnosed (4.7%), all in high-risk women. Three were diagnosed at the prevalence screen. Three were detected mammographically at subsequent screening rounds; three were detected by breast self-examination (BSE) between screening episodes. One interval cancer was visible on mammogram at presentation but not at screening five months previously. A second cancer was also visible on mammogram at presentation but the normal screening mammogram had been 17 months earlier, outside the recommended interval. The remaining interval cancer was not visible on the mammogram. A total of 363 two-view screening mammograms were performed in the 280 person-years of follow-up; 109 additional investigations were generated: 23 recall mammograms, 18 symptomatic mammograms, 45 ultrasounds, 12 aspiration cytologies and 11 biopsies. Cytology diagnosed malignancy in 1 of 12 cases; breast biopsy in 9 of 11 cases. Twenty-three additional women had ultrasound screening only. This audit suggests that screening below the age 50 years may be unnecessary in families with a low chance of having a BRCA1 or -2 mutation, but it is important to screen high-risk women at least annually and possibly under 35 years. PMID- 15131404 TI - Intron 4 mutation in APC gene results in splice defect and attenuated FAP phenotype. AB - The adenomatous polyposis coli (APC) protein is a tumor suppressor frequently involved in the development of inherited and sporadic colon cancers. Somatic mutations of the APC gene are found in 80% of all colon cancers. Inherited mutations result in familial adenomatous polyposis (FAP) as well as an attenuated form of this syndrome. FAP is characterized by the early age onset of hundreds to thousands of colonic adenomatous polyps and a virtual certainty of colon cancer unless the colon is removed. The attenuated form of FAP (AFAP) is characterized by fewer adenomas, later onset of adenomas and cancer, and a decreased lifetime cancer risk. We report a 37-year-old man with a history of more than 50 colonic adenomatous polyps, located predominately in the right colon. An insertion of a single thymidine between the second and third base pairs of intron 4 of the APC gene was identified (c.531+2_531+3insT). Monoallelic hybrid cells harboring a single copy of human chromosome 5 were generated from patient lymphoblasts. Sequencing of the APC cDNA product from these cells revealed a single RNA transcript with aberrant splicing in the mutant mRNA whereby exon 4 is deleted. The translational reading frame is shifted after codon 140 and a translational stop is generated predicting a truncated protein of 147 amino acids, thus indicating that the intronic mutation is disease causing. The lack of a secondary transcript from the mutant allele suggests that incomplete exon skipping is not the molecular mechanism behind the attenuated phenotype. PMID- 15131405 TI - Frequency of extra-colonic tumors in hereditary nonpolyposis colorectal cancer (HNPCC) and familial colorectal cancer (FCC) Brazilian families: An analysis by a Brazilian Hereditary Colorectal Cancer Institutional Registry. AB - The two main forms of hereditary colorectal cancer are familial adenomatous polyposis (FAP) and hereditary nonpolyposis colorectal cancer (HNPCC). Some families do not meet all the diagnostic criteria for HNPCC or FAP and are classified as familial colorectal cancer (FCC). Little information is available on the spectrum of tumors related to HNPCC and FCC in South America. OBJECTIVE: To describe the frequency of malignant tumors in a group of Brazilian families with HNPCC or FCC in an Institutional Hereditary Colorectal Cancer Registry. MATERIAL AND METHODS: The study included 61 families (29 HNPCC and 32 FCC) between January 1998 and June 2001. HNPCC families were clinically classified according to the Amsterdam Criteria I or II. FCC families were characterized by the presence of at least two individuals with CRC or extra-colonic tumors associated with the HNPCC spectrum, at least one of them being under 50 years of age. RESULTS: In the 29 families with HNPCC, 201 patients with cancer were identified among 1241 individuals (589 men and 652 women). Among the 201 patients 223 tumors were observed: 137 CRC (55 in men and 82 in women) and 86 extra colonic (37 in men and 49 in women). In the 32 families with FCC, 146 patients with cancer were identified among 1053 individuals (505 men and 548 women); 158 tumors were observed in 146 patients, 75 CRC (33 in men and 42 in women) and 83 extra-colonic tumors (47 in men and 36 in women). The most frequent extra-colonic primary sites among the HNPCC families were: endometrium (26.5%) and breast (26.5%) (women), and stomach (35.1%) (men). Among the FCC families, the most common primary sites were: breast (27.8%) (women), and stomach (44.4%) (men). CONCLUSION: The high frequency of endometrial and gastric cancer found was expected, since these tumors are part of the HNPCC spectrum, but the high frequency of breast cancer requires further molecular investigation to determine a possible hereditary predisposition associated with hereditary CRC. PMID- 15131406 TI - Patients' and professionals' opinions of services for people at an increased risk of colorectal cancer: an exploratory qualitative study. AB - OBJECTIVES: To explore the views and opinions of patients with a family history of colorectal cancer, and of primary and secondary care health professionals, on how to improve current services for individuals with a family history of colorectal cancer. DESIGN: Focus group and interview study. SETTING: Primary and secondary care centres in south east London, UK. PARTICIPANTS: A total of 53 people participated in this qualitative study: 18 individuals with a family history of colorectal cancer were interviewed in three focus groups and 35 health professionals were interviewed either in focus groups or individually. Visual qualitative analysis of transcribed interviews was used to identify the key themes which emerged. MAIN OUTCOME MEASURES: Interviewees' needs, views and opinions of current services and ways to improve them. RESULTS: Several areas of concern were identified. The role of primary care needs to be clarified. Education and information about services should be provided for patients and caregivers and better support is required for those undergoing screening and surveillance. Methods to ensure effective and meaningful risk communication are inadequate and require further exploration. Standardisation of care is required to ensure consistency of advice and treatment. CONCLUSION: These interviews revealed substantial shortfalls in the provision of services for patients at increased genetic risk of colorectal cancer. Current systems for the assessment of risk, delivery of advice, and for surveillance are inconsistent and sometimes maybe inadequate. The role of primary care physicians in service delivery requires clarification. Significant opportunities exist for the development of new, more appropriate models of service to provide better standards of care. PMID- 15131407 TI - Delivering information about cancer genetics via letter to patients at low and moderate risk of familial cancer: a pilot study in Wales. AB - Increasing demands upon specialist cancer genetics services have resulted in a need to explore alternative means of delivering genetic risk information to individuals at low-risk of familial cancer. This pilot study investigates patient satisfaction with a letter to low and moderate risk individuals notifying them of their risk. Sixty-six people completed a questionnaire designed to measure satisfaction with the way they had been notified of their cancer risk. Two key findings emerge from the data: first of all, whilst many respondents indicated overall satisfaction with the risk letter, a substantial number wanted more information about their risk; and secondly, low-risk individuals in this study are less reassured by and less satisfied with the risk letter than those at moderate risk. The optimal service provision for delivery of genetic risk information is likely to be one which can best respond to individual differences in information-seeking, distress and risk comprehension. There is a need therefore, for a randomised control trial to compare the effectiveness of a risk notification letter with more traditional telephone risk counselling and the implications of each mode of delivery upon the resources of specialist cancer genetics services. PMID- 15131408 TI - Health economics and genetic service development: a familial cancer genetic example. AB - Advances in genetics are allowing a greater number of clinical genetics services to be offered to individuals with family histories of particular diseases. It is important that such services are not allowed to proliferate before they have been properly evaluated in terms of both their clinical and cost-effectiveness. A randomised controlled study of a new clinical genetics service in Wales for women with a family history of breast cancer has been undertaken. The health service costs from initial consultation and counselling through to mutation testing within families were assessed, along with patient travel costs. The extra cost of specialist genetics assessment (intervention) was pound 16.36 higher per woman than assessment by a breast surgeon at a District General Hospital (control). A further cost of pound 46.42 per initially presenting woman was also generated as a result of further counselling within the families of 48, and testing within the families of 36 of the 412 original presenting women. An explanation is given here of the costing methodology as an illustration of what is involved in such an exercise. PMID- 15131410 TI - Familial cancer 2003 - research and practice, Couran Cove, Queensland, Australia, 3-6 September 2003. PMID- 15131409 TI - Familial pancreatic cancer. AB - The dismal prognosis of ductal pancreatic adenocarcinoma is mainly attributable to advanced tumor stages at the time of diagnosis. Meanwhile, familial pancreatic cancer is an established hereditary tumor entity that is responsible for approximately 3% of pancreatic cancer (PC) cases. Therefore, analysis of the family history may help to identify individuals at increased risk of developing PC. These include members of families with a history of PC as well as those of families with distinct hereditary cancer syndromes such as Peutz-Jeghers syndrome, hereditary pancreatitis, familial atypical multiple mole melanoma syndrome, hereditary breast and ovarian cancer syndrome and hereditary non polyposis colorectal cancer. In future, the identification of germline mutations in genes predisposing to PC, together with the analysis of exogenous risk factors, could be used for a more precise risk assessment for the development of PC. This may allow the application of invasive screening methods for the identification of early PC or, even better, its precursor lesions in high-risk individuals, providing the option of timely curative pancreatectomy. PMID- 15131412 TI - Nurses in the media: where are home care and hospice? PMID- 15131414 TI - Are you salt savvy? PMID- 15131415 TI - M0100--Reason for Assessment (RFA): revised and updated. PMID- 15131416 TI - How MedPac impacts home care. PMID- 15131417 TI - Involving patients in care decisions improves satisfaction: an outcomes-based quality improvement project. AB - A home care agency used quality improvement processes to improve patient satisfaction survey ratings. The focus was on involving patients in decisions about their care. A multidisciplinary team developed creative strategies to increase staff awareness and enhance customer service skills, which had dramatic results. PMID- 15131418 TI - Building the foundations of wound care training: the need to fight the mantra "time is money". AB - The home care wound patient is continuing to grow and requires a significant organizational commitment to keep field staff current. One agency's successful effort to develop a wound management program is presented. PMID- 15131419 TI - The right assessments = the right PPS payment. PMID- 15131422 TI - Back to the basics: the clinical record. AB - This article identifies the various functions and outlines the requirements of the clinical record in home care. PMID- 15131424 TI - A 10-step path for conflict resolution. PMID- 15131425 TI - Understanding the impact of the new Medicare law on home health patients--part 1. PMID- 15131426 TI - Levothyroxine and calcium interaction: timing is everything. AB - Are you aware that clients taking levothyroxine (Levoxyl, Synthroid, Unithroid) should also be taking a calcium supplement and that the calcium supplement should be spaced at least 4 hours apart from levothyroxine? PMID- 15131427 TI - Artificial nutrition and hydration in end-of-life care. HPNA position paper. PMID- 15131428 TI - What exactly is a "real nurse?". PMID- 15131429 TI - Presidential address: The formation of ISSLS and its impact on lumbar spine research. PMID- 15131430 TI - A randomized study of closed wound suction drainage for extensive lumbar spine surgery. AB - STUDY DESIGN: A prospective randomized study. OBJECTIVES: To study the risk of infection, hematoma, and neurologic deficits following extensive lumbar spine surgery in patients with or without prophylactic closed wound suction drain placement. SUMMARY OF BACKGROUND DATA: One randomized study assessing prophylactic drain placement in one-level lumbar spine surgery suggested that the use of a wound drain is not effective at preventing infection and may actually increase the rate of this complication. Our study was designed to determine the efficacy of closed wound suction drainage in preventing complications after extensive lumbar spine surgery. METHODS: Eighty-three consecutive patients undergoing extensive lumbar spine surgery were prospectively randomized to one of two groups. Forty-two patients had a closed wound suction drain placed before wound closure and 41 patients did not have a drain placed. The two groups were then assessed for differences in postoperative infection rate, incidence of hematoma and neurologic deficits, operating room time, estimated blood loss, hemoglobin and hematocrit values, temperature, dressing drainage, and length of hospital stay. RESULTS.: No infections, epidural hematomas, or new neurologic deficits were encountered in either group of patients. The only significant finding was a higher temperature in the "no drain" group the first day after surgery (P = 0.0437). CONCLUSIONS: Based on the findings in this and other studies, the decision to use or not use a wound drain following lumbar spine surgery should be left to the surgeon's discretion. PMID- 15131431 TI - Mini-intervention for subacute low back pain: two-year follow-up and modifiers of effectiveness. AB - STUDY DESIGN: Randomized controlled trial. OBJECTIVES: To Investigate the long term effectiveness, costs, and effect modifiers of a mini-intervention, provided in addition to the usual care, and the incremental effect of a worksite visit for patients with subacute disabling low back pain (LBP). SUMMARY OF BACKGROUND DATA: A mini-intervention was earlier proved to be an effective treatment for subacute LBP. Whether the beneficial effect is sustained is not known. Furthermore, modifiers of a treatment effect are largely unknown. METHODS: A total of 164 patients with subacute LBP randomized into a mini-intervention (A, n = 56), a mini-intervention plus a worksite visit (B, n = 51), or the usual care (C, n = 57). Mini-intervention consisted of a detailed assessment of the patients' history, beliefs, and physical findings by a physician and a physiotherapist, followed by recommendations and advice. The usual care patients received the conventional care. Pain, disability, health-related quality of life, satisfaction with care, days on sick leave, and health care consumption and costs were measured during a 24-month follow-up. Thirteen candidate modifiers were tested for each outcome. RESULTS: There were no differences between the three treatment arms regarding the intensity of pain, the perceived disability, or the health related quality of life. However, mini-intervention decreased occurrence of daily (A vs., C, P = 0.01) and bothersome (A vs. C, P < 0.05) pain and increased treatment satisfaction. Costs resulting from LBP were lower in the intervention groups (A 4670 Euros, B 5990 Euros) than in C (C 9510 Euros) (A vs. C, P = 0.04; and B vs. C, not significant). The average number of days on sick leave was 30 in A, 45 in B, and 62 in C (A vs. C, P = 0.03; B vs. C, not significant). The perceived risk for not recovering was the strongest modifier of treatment effect. Mental and mental-physical workers in A and B were less often on sick leave than those in C. CONCLUSIONS: Mini-intervention is an effective treatment for subacute LBP. Despite lack of a significant effect on intensity of low back pain and perceived disability, mini-intervention, including proper recommendations and advice, according to the "active approach," is able to reduce LBP-related costs. The perceived risk of not recovering was the strongest modifier of treatment effect. In alleviating pain, the intervention was most effective among the patients with a high perceived risk of not recovering. PMID- 15131432 TI - Innervation of the lumbar intervertebral disc by nerve growth factor-dependent neurons related to inflammatory pain. AB - STUDY DESIGN: We used anatomic tracers and immunoreactivity in rats to define dorsal root ganglion neuron populations innervating the lumbar discs in physiologic and inflammatory states. OBJECTIVES: To investigate the percentages of calcitonin gene-related peptide-immunoreactive (CGRP-ir) and isolectin B4 (IB4)-binding neurons innervating lumbar discs. SUMMARY OF BACKGROUND DATA: Small neurons are classified into two types. One contains CGRP and expresses the nerve growth factor receptor. The other binds IB4 and expresses the glial cell line derived neurotrophic factor receptor. METHODS: A neurotracer, Fluoro-Gold, was applied to the L5-L6 disc in rats. Five days later, 50-microL saline (control group: n = 8) or Complete Freund's adjuvant (inflammatory group: n = 8) was applied to the disc. Seven days after the second operation, T13-L5 dorsal root ganglions were processed for double staining of CGRP and IB4. RESULTS: Of the Fluoro-Gold-labeled neurons, 50.1 +/- 4.6% (mean +/- SEM) were positive for CGRP and 0.7 +/- 0.6% positive for IB4 in the control group, while 65.6 +/- 4.7% were positive for CGRP and 1.0 +/- 1.0% positive for IB4 in the inflammatory group. The percentage of CGRP-ir neurons was significantly higher than that of IB4 binding neurons in both groups (P < 0.001, each). The percentage of CGRP-ir neurons in the inflammatory group was significantly higher than in the control group (P < 0.05). CONCLUSIONS: We found that most small neurons innervating the disc were CGRP-ir. Furthermore, disc inflammation caused an increase in CGRP-ir neurons but not IB4-binding neurons, suggesting that CGRP-ir, nerve growth factor dependent neurons are more responsible for discogenic pain. PMID- 15131433 TI - TNF-alpha and TNF-alpha receptor type 1 upregulation in glia and neurons after peripheral nerve injury: studies in murine DRG and spinal cord. AB - OBJECTIVES: The purpose of the current study was to evaluate changes in tumor necrosis factor-alpha (TNF-alpha ) and TNF-alpha receptor 1 (p55 receptor) using double fluorescent immunohistochemistry in glial and neural cells in the dorsal root ganglion and spinal cord after sciatic nerve injury in mice. SUMMARY OF BACKGROUND DATA.: TNF-alpha is a primary mediator of the inflammatory response and is primarily synthesized and released in the nervous system by macrophages and Schwann cells following peripheral nerve injury. TNF-alpha is also released from astrocytes and microglia in the central nervous system, where it plays a crucial role in the pathophysiology of injury. METHODS: Sixteen female mice were used. Under anesthesia, the left sciatic nerve was crushed. At 3, 5, and 14 days after surgery, the spinal cord at the level of L5 and the left L5 DRG were removed and processed for immunohistochemistry. Tissue sections were double stained with antibodies to either glial fibrillary acidic protein (GFAP; marker for astrocytes or satellite cells) or NeuN (marker for neurons), and TNF or p55 receptor. RESULTS.: In the dorsal root ganglion, GFAP-immunoreactive (IR) satellite cells became evident after injury and were also immunoreactive for both TNF-alpha and p55 receptor. Dorsal root ganglion neurons expressed p55 receptor after injury. TNF-alpha and GFAP-IR satellite cells surrounded p55-IR neurons. Furthermore, the number of GFAP-IR astrocytes dramatically increased in the spinal cord after nerve injury, and some astrocytes were also TNF-alpha -IR and p55 receptor-IR. TNF-alpha -1R astrocytes were seen around p55 receptor-IR neurons. CONCLUSIONS: These data demonstrate that upregulation of glial TNF-alpha is associated with the expression of the p55 receptor on adjacent neurons. This association may have induced the expression of several cytokines and immediate early genes in dorsal root ganglion and spinal cord neurons via the TNF signaling pathway. These findings may be related to the pathogenesis of neuropathic pain. PMID- 15131434 TI - Interleukin-6 production is upregulated by interaction between disc tissue and macrophages. AB - STUDY DESIGN: Interleukin (IL)-6 production was investigated using a coculture system of disc tissue and macrophages. OBJECTIVES: The purpose of this study was to investigate the interaction between intervertebral disc tissue and macrophages in terms of IL-6 production. SUMMARY OF BACKGROUND DATA: IL-6 production is observed in human herniated disc specimens, and there is a correlation between IL 6 production and neurologic symptoms. However, the mechanism of IL-6 production in the herniated disc is not clear. MATERIALS AND METHODS: Coccygeal intervertebral discs and exudated peritoneal macrophages were obtained from male Sprague-Dawley rats. Macrophages and intervertebral disc without endplates were cocultured in a serum-free medium. Fat tissue culture with or without macrophages, intervertebral disc alone, and macrophages alone were used for controls. The supernatant fluid of the culture was utilized for the enzyme-linked immunosorbent assay. The precipitations of macrophages and disc coculture were used for semiquantitative RT-PCR for IL-6. Immunohistochemical staining for IL-6 and the macrophages marker (ED2) were also carried out using disc tissue cultured with macrophages. RESULTS: IL-6 production level was significantly increased in the coculture of intervertebral disc and macrophages (P < 0.01). However, there was no significant production of IL-6 in the control groups. The precipitations from coculture of macrophages and disc expressed IL-6 mRNA in semiquantitative RT PCR. Immunohistochemical staining revealed most IL-6 producing cells were also positive for ED2, which adheres to or infiltrates the peripheral area of the nucleus pulposus. CONCLUSIONS: Our results demonstrated that interaction between disc tissue and macrophage is necessary for upregulation of IL-6 production. Immunohistochemical staining also indicated that infiltrated macrophages played a major role in production of IL-6, suggesting that infiltration of macrophages into herniated disc material may be a trigger for IL-6 production and associated neurologic symptoms. PMID- 15131436 TI - Altered disc mechanics in mice genetically engineered for reduced type I collagen. AB - STUDY DESIGN: Mechanically test lumbar discs of transgenic mice in compression tension and torsion. OBJECTIVES: Determine if a reduction in type I collagen results in decreased disc mechanics. SUMMARY OF BACKGROUND DATA: Quantitative relationships between disc structure and function would improve the understanding of disc generation and are essential relationships for functional tissue engineering. The reduced type I collagen transgenic mouse has been used in structure-function studies of bone and tendon, but not intervertebral discs. Methods for testing mouse discs have recently been developed, making disc structure-function studies possible. METHODS: Microradiographed and mechanically tested lumbar discs from control and collagen-reduced mice in both compression tension and torsion were used. Disc area and polar moment of inertia were determined from radiographic data, stiffness from mechanical data, and apparent modulus from geometric and mechanical data. RESULTS: Collagen-reduced discs had a larger area and polar moment of inertia compared to controls. The linear and torsional stiffness of collagen-reduced and control discs were not significantly different. Finally, the apparent modulus of collagen-reduced discs was significantly less than controls in compression (73% of control) and torsion (50%). CONCLUSIONS: Compared to controls, collagen-reduced discs had reduced apparent modulus in both loading directions, suggesting that the transgenic disc tissue was mechanically inferior to controls. These results are consistent with the widely accepted functional role of type I collagen in disc mechanics, and therefore supports the use of transgenic mice to study structure-function relationships of the disc. Future work will focus on quantifying structure function relationships related to degeneration, as well as those relevant to the design of tissue-engineered disc replacements. PMID- 15131437 TI - The functional significance of cell clusters in the notochordal nucleus pulposus: survival and signaling in the canine intervertebral disc. AB - STUDY DESIGN: Cell viability was assessed in relation to cell clustering, and mechanisms of cell-cell signaling in the clusters were investigated. OBJECTIVES: To explore the functional role of cell clustering in the notochordal nucleus pulposus. SUMMARY OF BACKGROUND DATA: The intervertebral disc of some species contains residual cells from the embryonic notochord. These cells form large three-dimensional clusters in the young, healthy disc but are replaced by chondrocyte-like cells during aging and degeneration. METHODS: Forty nucleus pulposi of adult dog lumbar intervertebral discs were isolated, and were left untreated, mechanically disrupted through a syringe, or enzymatically digested. The presence of functional gap junctions was determined by the fluorescence recovery after photobleaching method. Cell viability was also assessed over 20 days in vitro. RESULTS: The cell clusters were interconnected via functional gap junctions. Mechanical disruption of the tissue had little effect on long-term cell viability, but enzymatic disruption of the tissue had a substantial negative impact on cell survival. CONCLUSIONS: These results demonstrate that the notochordal cells in adult dog nucleus pulposi are able to communicate via cytoplasmic signals and that such communications may influence the functionality of these cells in the young disc. PMID- 15131438 TI - Experimental spinal stenosis: relationship between degree of cauda equina compression, neuropathology, and pain. AB - STUDY DESIGN: An analysis of pathologic changes after different degrees of cauda equina compression. OBJECTIVES: To explore the association between the degree of the cauda equina compression and the extent of pathologic change, expression of tumor necrosis factor (TNF-alpha), and neuropathic pain. To compare with distal nerve compression injury. SUMMARY OF BACKGROUND DATA: Compression of the cauda equina reduces blood flow in compressed nerve roots and causes TNF-alpha expression and neuropathological change. In peripheral nerve, expression of TNF alpha in Schwann cells is associated with primary demyelination without pain while TNF-alpha expression by macrophages is associated with axonal (Wallerian) degeneration and pain. METHODS: Two square-shaped pieces of silicon were placed into the fourth and sixth epidural space in rats. Various sized silicon was used in each group (mild, moderate, and strong compression groups), while no silicon was used in the sham-operated group. Mechanical allodynia was determined by the von Frey test. Comparisons of the number of TNF-alpha- and apoptosis-positive cells were made using immunohistochemistry. RESULTS: There was no significant mechanical allodynia observed in any group. Some nerve roots showed demyelination following mild cauda equina compression. Axonal degeneration was observed in the moderate and strong cauda equina compression groups. TNF-alpha-immunoreactive cells were increased in all compression groups. Apoptosis of dorsal root ganglion cells was less than apoptosis in the spinal cord. CONCLUSION: Mild cauda equina compression induces TNF-alpha expression and demyelination. Moderate and strong cauda equina compression induces TNF-alpha expression and degeneration associated with macrophage invasion. Neither demyelination nor degeneration in the cauda equina induced mechanical allodynia. Nerve lesions proximal to the dorsal root ganglion do not produce significant mechanical allodynia. Dorsal root ganglion apoptosis may be important for pain. PMID- 15131439 TI - Prospective controlled study of the development of lower back pain in previously asymptomatic subjects undergoing experimental discography. AB - STUDY DESIGN: A prospective controlled longitudinal study. OBJECTIVES: To determine whether subjects, asymptomatic for lower back problems, who undergo experimental discography, will develop lower back problems during the medium term to the full term. SUMMARY OF BACKGROUND DATA: Previous work has shown significant pain on discographic injection in approximately 40% of asymptomatic subjects. It has been suggested that those subjects with painful injections would soon develop lower back pain (LBP) syndromes in the near future: that is, the experimental discography was detecting an imminent "pain generator" before clinically symptomatic. METHODS.: Fifty subjects without low back pain were recruited for clinical and psychometric testing, MRI scanning, and experimental lumbar discography to determine the rate of painful lumbar disc injections in select subjects without LBP history. After determining which subjects had painful injections, all subjects completing the discography protocol were prospectively followed at yearly intervals to determine the occurrence of LBP and LBP disability over time. Statistical methods were then used to determine the correlation, if any, between the asymptomatic subjects' clinical, MRI, and discography findings, and the subsequent LBP measures. Controls, not participating in the lumbar discography study, were also followed. Controls were matched for clinical features, sex, age, and occupational/recreational exposure. Follow-up examinations were performed at yearly intervals by blinded researchers using a scripted interview and completing standard questionnaires. RESULTS: A total of 46 of 50 completed the discogram, and all 46 subjects completed the final 4-year follow-up examination. There was a low incidence of LBP episodes in the experimental groups and control. A painful disc injection, independent of psychological profile, did not predict LBP or any other functional outcome measure at follow-up on multivariate analysis. The presence of an anular fissure seen on discography was weakly associated with the cumulative incidence of LBP episodes after discography (P = 0.08). The presence of high intensity zone on MRI in any disc was also weakly associated with the development of LBP episodes (P = 0.09). Psychometric profiles at the start of the study strongly and independently predicted future back pain (P = 0.01), medication usage (P = 0.002), and work loss (P = 0.01) over the 4-year study. Compared with controls not having undergone discography, there was no significant difference in back pain, function, work loss, doctors visits for back pain, or medication intake in any group. A subset in the injection group with somatization disorder had a higher LBP visual analog score compared with somatization disorder controls at 1 year,but this was not significant at 4 years after testing. CONCLUSIONS: Painful disc injections are poor independent predictors of subsequent LBP episodes in subjects initially without active lower back complaints. Anular disruption is a weak predictor of future LBP problems. Psychological distress and preexisting chronic pain processes are stronger predictors of LBP outcomes. PMID- 15131440 TI - Comparison of methods for determining the presence and extent of anterior lumbar interbody fusion. AB - STUDY DESIGN: Titanium alloy interbody fusion devices with autogenous bone were placed in the L5-L6 disc space of 31 adult pig-tailed monkeys through an anterolateral (retroperitoneal) approach. Anteroposterior and lateral radiographs, CT imaging, and histologic analysis of the specimens were performed. OBJECTIVES: This study compared the accuracy of plain film radiographs and CT imaging for determining bony fusion of a titanium interbody device implanted in a non-human primate model. The accuracy of the assessments was determined by comparison to histologic analysis. SUMMARY OF BACKGROUND DATA: Interbody fusion assessment is often difficult to compare in clinical studies because of differences in definition of fusion criteria. In addition, the accuracy of plain film radiographs and CT imaging assessments of fusion are debated because of device material radiopacity and introduction of artifacts. METHODS: A uniform grading system evaluating both the presence and extent of bony fusion was applied to all evaluation techniques. Matched-pair nonparametric t tests were used to determine differences in scoring. RESULTS: The radiographic and histologic presence of fusion grades was equivalent in only 13 of 29 cases (45%), while the CT imaging was equivalent to histologic assessment in 24 of 29 cases (83%). However, the extent of bony fusion in CT imaging and histologic assessment was equivalent in only 4 of 29 cases (14%). Grading of CT images significantly overestimated the extent of fusion. CONCLUSIONS: This study demonstrated CT imaging techniques to be superior to plain film radiographs in determining the presence of bony fusion. However, CT imaging did not accurately determine the extent of bony fusion present as confirmed by histologic analysis. PMID- 15131441 TI - Temperatures within the lumbar disc and endplates during intradiscal electrothermal therapy: formulation of a predictive temperature map in relation to distance from the catheter. AB - STUDY DESIGN: Temperatures were measured in human cadaveric lumbar discs during intradiscal electrothermal therapy. OBJECTIVES: To determine if sufficient temperatures for collagen denaturation and nociceptive ablation can be achieved at clinically significant distances from the intradiscal electrothermal therapy heating catheter. SUMMARY OF BACKGROUND DATA: Previous cadaveric studies have contested the ability of the intradiscal electrothermal therapy catheter to produce sufficient temperatures to denature collagen and cause neural ablation within the posterior anulus of the disc. However, these experiments used clinically unrepresentative device placements along the anterolateral anulus. METHODS: Intradiscal electrothermal therapy was performed in 14 human cadaveric discs. Devices were inserted using a standard posterolateral approach to orient the heating catheter along the posterior anulus. Temperature recordings were collected using multiple sensors placed along the posterior anulus, anterior anulus, and endplates. RESULTS: Temperatures greater than 60 C and 65 C were achieved in 14 and 5 specimens, respectively, at distances up to 2 mm from the catheter. Between 2 and 4 mm, more than 60 C was achieved in all specimens. More than 45 C was achieved in all specimens at distances of 9 to 14 mm from the device. CONCLUSIONS: Temperatures sufficient for collagen denaturation and nociceptive ablation were detected at distances greater than previously documented. These data suggest that intradiscal electrothermal therapy's proposed heat-dependent mechanisms of action are achievable in most discs. Among other factors, interspecimen variability of maximum temperatures may help explain the somewhat inconsistent clinical results following intradiscal electrothermal therapy. PMID- 15131443 TI - Facet angles in lumbar disc herniation: their relation to anthropometric features. AB - STUDY DESIGN: Cross-sectional study. OBJECTIVE: To investigate the alteration of facet joint angle and its relation to anthropometric features in lumbar disc herniation. SUMMARY OF BACKGROUND DATA: In the previous studies, the facet tropism and its relation to the level, type, side, and development of lumbar disc herniation were reported. This study was the first to correlate anthropometric feature, facet angle, and asymmetry in patients with lumbar disc herniation. METHODS.: Sixty-one patients with lumbar disc herniation were included in this study. The body height, vertex-to-spina iliaca posterior superior distance, spina iliaca posterior superior-to-floor distance, and net weight were measured and body mass index calculated. The facet angles were measured for each facet joint using computed tomography. The intraobserver error was found to be +/- 1.66 degrees. We defined facet tropism as the bilateral angle difference greater than 2 intraobserver errors. The asymmetry rate was a ratio of degree of angles of the more coronally oriented facet joint to the more sagittally oriented facet joint at the same intervertebral level. RESULTS: The facet asymmetry was observed at the herniation level in 70.5% of the patients. There was a correlation between the degree of the more sagittally oriented facet joint angle and facet asymmetry rate at the herniation level (R = -0.684, P = 0.0001 for L4-L5 lumbar disc herniation; R = -0.509, P = 0.008 for L5-S1 lumbar disc herniation). The body height correlated with the degree of the more sagittally oriented facet joint angle, the degree of more coronally oriented facet joint angle, and asymmetry rate, respectively (R = -0.571, P = 0.0001; R = -0.474, P = 0.002; R = 0.314, P = 0.045, respectively). There was no correlation between the degree of these angles and the vertex-to-spina iliaca posterior superior distance. The spina iliaca posterior superior-to-floor distance was correlated with the degree of the more sagittally oriented facet joint angle (R = -0.457, P = 0.0001) and more coronally oriented facet joint angle (R = -0.435, P = 0.004) but not with the asymmetry rate. The degree of facet angles was not correlated with body weight and body mass index. CONCLUSION: This study revealed that patients with lumbar disc herniation had the asymmetry and sagittalization of facet joints, and these alterations were more evident in the taller patients. PMID- 15131444 TI - Systematic correlation of transcranial magnetic stimulation and magnetic resonance imaging in cervical spondylotic myelopathy. AB - STUDY DESIGN: A prospective study over a 3.5-year period involving transcranial magnetic stimulation and magnetic resonance imaging. OBJECTIVES: To assess the correlation of transcranial magnetic stimulation and magnetic resonance imaging in cervical spondylotic myelopathy qualitatively and statistically. SUMMARY OF BACKGROUND DATA: Cervical spondylotic myelopathy presents with different degrees of cord compression, which can be assessed by magnetic resonance imaging. There are no large studies correlating transcranial magnetic stimulation and magnetic resonance imaging findings in this condition. METHODS: A total of 141 patients with a clinical diagnosis of cervical spondylotic myelopathy were prospectively studied over a 3.5-year period. They were classified into Groups 1 to 4 based on severity of cervical cord changes on magnetic resonance imaging. All had transcranial magnetic stimulation and central motor conduction time measurements within 2 months of the magnetic resonance imaging study. RESULTS: Twenty-eight, 49, 28, and 36 patients were classified into Groups 1 to 4, respectively. Mean upper limb and lower limb central motor conduction times correlated with the severity of magnetic resonance cord compression. The absence of central motor conduction time abnormalities correlated reliably with the absence of cervical cord impingement as in Group 1. Statistically significant right left difference in central motor conduction time in the lower limbs was seen between Groups 1 (no cord changes) and Group 2 (mild cord impingement). Eight other patients with diagnoses other than cervical spondylotic myelopathy all showed central motor conduction time abnormalities. The sensitivity and specificity for transcranial magnetic stimulation for differentiating the presence from absence of magnetic resonance imaging cord abnormality were 100% and 84.8%, respectively. CONCLUSIONS: Transcranial magnetic stimulation showed excellent correlation with magnetic resonance imaging findings and can be considered as an effective technique for screening patients for cervical cord abnormalities before magnetic resonance imaging in the clinical setting. The findings in this study have relevant implications in the pathophysiology, management, and health costs of cervical spondylotic myelopathy. PMID- 15131445 TI - Type II error in the spine surgical literature. AB - STUDY DESIGN: A literature review. OBJECTIVES: To determine the frequency of potential type II errors published in the spine surgical literature. SUMMARY OF BACKGROUND DATA: The randomized controlled trial is the strongest clinical evidence available in investigational medicine. Unfortunately, it is common for randomized controlled trials published in peer-reviewed journals not to report a primary question or a sample size calculation. When the null hypothesis is accepted and the power of a study is unreported, the validity of a study's findings may be significantly limited. To our knowledge, the spine literature has not been appraised to determine the frequency of type II errors. METHODS.: A literature search was conducted of MED-LINE, PubMed, and Cochrane databases, using the key words of "spine" and "surgery" between 1967 and 2002. Trials were included if they were of a 2-group randomized controlled trial design, which reported a nonsignificant difference in the primary outcome. The frequency of reporting the primary outcome and sample size calculation was determined. The sample size was assessed to determine whether the trial had sufficient patients to detect a 10%, 25%, and 35% relative difference in the primary outcome for a power of 80%. RESULTS: A total of 37 studies satisfied the inclusion criteria. Six studies reported a sample size calculation (17%). Of the remaining 31 studies, 5 explicitly stated a primary outcome (14%). The mean type II error (beta error) was 82%. CONCLUSION: The spine surgical literature is plagued with a high potential for type II error. A trial's methodology should be scrutinized to prevent misinterpretation of the results. PMID- 15131446 TI - Anterior spinal arthrodesis with structural cortical allografts and instrumentation for spine tumor surgery. AB - STUDY DESIGN: The authors report on anterior vertebral reconstruction following tumor resection with use of fresh-frozen, cortical, long-segment allografts prepared from diaphyseal sections of long bones. A retrospective analysis of clinical outcomes is presented. OBJECTIVE: To analyze the results following the use of cortical allografts in the treatment of spine tumors. SUMMARY OF BACKGROUND DATA: Metastatic disease and primary spinal bone tumors may result in progressive vertebral collapse, instability, deformity, pain, and neurologic deficit. Controversy as to the appropriate type of anterior reconstruction and/or graft material persists. METHODS: From 1995 until 2001, 30 patients with primary spinal bone tumors or metastases to the spine were treated by anterior vertebral reconstruction with fresh-frozen cortical bone allografts. Grafts were used in combination with anterior and posterior instrumentation. RESULTS: The median survival was 14 months. Ninety-three percent of all allografts were radiographically incorporated as early as 6 months after surgery in spite of adjuvant chemotherapy and radiation therapy. Fourteen patients (46%) had intraoperative or postoperative complications. Two patients underwent revision surgery for local recurrence. There were no allograft infections, fractures, or collapse. CONCLUSION: Anterior column reconstruction with structural cortical allografts proved to be a reliable technique in patients with spine tumors. Postoperative complications can often be successfully managed. PMID- 15131448 TI - Re: Brox JI, Sorensen R, Friis A, et al. Randomized clinical trial of lumbar instrumented fusion and cognitive intervention and exercises in patients with chronic low back pain and disc degeneration. Spine. 2003; 28:1913-1921. PMID- 15131450 TI - Re: Belkoff SM, Molloy S. Temperature measurement during polymerization of polymethyl methacrylate cement used for vertebroplasty. Spine. 2003; 28:1555-9. PMID- 15131451 TI - Re: Weiner BK, Walker M. Autologous growth factors and lumbar intertransverse fusions. Spine. 2003; 28:1968-70; discussion 1971. PMID- 15131452 TI - Re: Weiner BK, Walker M. Autologous growth factors and lumbar intertransverse fusions. Spine 2003;28: 1968-70; discussion 1971. PMID- 15131454 TI - The efficacy of a treatment program focusing on specific stabilizing exercises for pelvic girdle pain after pregnancy: a two-year follow-up of a randomized clinical trial. AB - STUDY DESIGN: A randomized clinical trial. OBJECTIVES: To examine the effects of a treatment program focusing on specific stabilizing exercises after a 2-year follow-up period. SUMMARY OF BACKGROUND DATA: An individualized treatment approach with specific stabilizing exercises is shown to be effective for women with pelvic girdle pain 1 year after delivery. No previous study has examined the long-term effects of treatment for women with postpartum pelvic girdle pain. METHODS: Eighty-one women with pelvic girdle pain postpartum were assigned randomly to 2 treatment groups for 20 weeks. Patient self-reported questionnaires measuring pain, disability, and health-related quality of life were collected after 20 weeks of treatment and 1 and 2 years postpartum. RESULTS: All 81 women returned the questionnaires for the 2-year follow-up. Sixteen were excluded from the analysis, mainly due to new pregnancies. The significant differences between the groups in functional status, pain, and physical health (SF-36) were maintained 2 years after delivery. Minimal disability was found in 85% of the specific stabilizing exercise group as compared to 47% in the control group. The control group showed significant improvement in functional status with median change score of 6.0 (Q1-Q3 of -12-0). Minimal evening pain was reported by 68% in the specific stabilizing exercise group versus 23% in the control group. However, the group differences disappeared for all measures when controlling for score level 1 year after delivery by regression analysis. CONCLUSION: The significant differences between the groups persisted with continued low levels of pain and disability in the specific stabilizing exercise group 2 years after delivery. Significant reduction in disability was found within the control group. Those with the highest level of disability and greatest potential for improvements recovered most, regardless of intervention group. PMID- 15131455 TI - Measurement of occipitocervical angle. AB - STUDY DESIGN: This study compared the reliability of 3 techniques used to measure alignment between the occiput and cervical spine. OBJECTIVES: Intraobserver and interobserver intraclass correlation coefficient were computed to determine the most reliable method to measure occipitocervical angle. SUMMARY OF BACKGROUND DATA: No studies have been performed comparing occipitocervical angle measurement techniques. METHODS: The angles between the inferior endplate of second cervical vertebrae and the occiput line using the Chamberlain line, McRae line, and McGregor line were measured from lateral cervical radiographs of 30 healthy volunteers. Five spine surgeons made measurements. RESULTS: Mean intraobserver variances of the angles using Chamberlain line, McRae line, and McGregor line were 2.0 degrees (ranging from 0 degrees-15 degrees), 4.7 degrees (from 0 degrees 28 degrees), and 1.5 degrees (from 0 degrees-9 degrees), respectively; intraobserver intraclass correlation coefficients of the angles were 0.956, 0.835, and 0.975. Mean interobserver variances of the angles using Chamberlain line, McRae line, and McGregor line were 2.3 degrees (from 0.4 degrees-6.4 degrees), 5.0 degrees (from 1.8 degrees-11.9 degrees), and 1.4 degrees (from 0 degrees-4.5 degrees), respectively; interobserver intraclass correlation coefficients were 0.939, 0.802, and 0.972. The highest reliability indexes were obtained for McGregor line. CONCLUSIONS: The McGregor line is the most reproducible and reliable method for measurement of the occipitocervical angle. PMID- 15131456 TI - Serious pyogenic spondylitis following vertebroplasty--a case report. AB - STUDY DESIGN: The aim of this case report is to examine the problem of serious pyogenic spondylitis that may follow vertebroplasty. OBJECTIVES: To report 1 case of serious pyogenic spondylitis caused by percutaneous vertebroplasty with polymethyl methacrylate. SUMMARY OF BACKGROUND DATA: Serious pyogenic spondylitis caused by percutaneous vertebroplasty with polymethyl methacrylate is a rare complication. We herein report a case of pyogenic spondylitis that occurred after the patient had undergone vertebroplasty. METHODS: A 78-year-old woman with a T12 osteoporotic compression fracture, and without a magnetic resonance imaging sign of infection, underwent percutaneous vertebroplasty using polymethyl methacrylate without complication. RESULTS.: About 1 month after the operation, the patient was readmitted because of severe backache, and the diagnosis of pyogenic spondylitis was made using radiograph and magnetic resonance imaging. Anterior inter-body fusion with a strut bone graft after debridement and posterior instrumentation were then performed. CONCLUSIONS: Vertebroplasty should proceed under sterile conditions. Excluding the possibility of spinal infection before surgery and a detailed evaluation assuring that the patient is without systemic infectious disease before the vertebroplasty procedure is crucial. PMID- 15131459 TI - Consensus: varicella vaccination of healthy children--a challenge for Europe. AB - The seriousness of varicella-zoster virus (VZV) infection as a public health issue is becoming clearer as country-specific epidemiologic and pharmacoeconomic data become available. In Germany, for example, studies have shown that >5.5% of immunologically healthy individuals develop varicella-related complications such as bacterial superinfections, acute neurologic disorders, pneumonia, bronchitis and otitis media; whereas in Italy, 3.5 to 5% of childhood cases of varicella cause complications such as upper respiratory tract and cutaneous infections. Varicella vaccines are now available. These live attenuated Oka strain vaccines have been shown in extensive studies to be highly immunogenic and well-tolerated in immunocompetent and immunocompromised children, with seroconversion rates ranging from 94 to 100% and 53 to 100%, respectively. These vaccines are also highly effective against clinical disease. These considerations led to a reevaluation of varicella vaccination policies. A routine varicella vaccination program targeting healthy children has already been implemented in the US, and data produced are encouraging and valuable. Similar strategies have not yet been adopted across Europe. The European Working Group on Varicella (EuroVar) was formed in 1998 to address the issues surrounding varicella epidemiology in Europe. After a series of meetings, the EuroVar members prepared a consensus statement recommending routine varicella vaccination for all healthy children between 12 and 18 months and to all susceptible children before their 13th birthday, in addition to catch-up vaccination in older children and adults who have no reliable history of varicella and who are at high risk of transmission and exposure. However, such a policy is recommended only if a very high coverage rate can be achieved. This could be reached with a measles-mumps-rubella varicella combined vaccine. PMID- 15131460 TI - Potential role of fluoroquinolone therapy in childhood otitis media. AB - Increased resistance of pneumococci and other pathogens to available antibiotics raises concerns about bacteriologic and clinical failure in children with acute otitis media. Few therapeutic options exist for patients with recurrent infections or recent treatment failure. The limited experience from the compassionate use of fluoroquinolones in pediatrics and pediatric studies has not been linked unequivocally with arthrotoxicity, the primary safety concern in children. Newer 8-methoxyfluoroquinolones may have a role in selected cases associated with multidrug-resistant pathogens. PMID- 15131461 TI - Azithromycin compared with beta-lactam antibiotic treatment failures in pneumococcal infections of children. AB - OBJECTIVE: To determine whether treatment failures occurred more commonly with azithromycin than with beta-lactam antibiotics in children who developed invasive pneumococcal disease within 30 days of receiving prior antimicrobial therapy. METHODS: Retrospective review of medical records of children evaluated at Texas Children's Hospital between 1996 and 2002 who had received antimicrobials (azithromycin or a beta-lactam antibiotic) and developed invasive pneumococcal disease within 30 days. Treatment failure was defined as invasive pneumococcal infection that occurred while taking antimicrobials or within 3 days of stopping azithromycin treatment or 1 day of stopping beta-lactam treatment. Penicillin and azithromycin susceptibilities were determined and categorized according to National Committee for Clinical Laboratory Standards guidelines. RESULTS: We identified 21 and 33 children with similar demographic features who had developed invasive pneumococcal disease within 1 month of receiving azithromycin or a beta lactam antibiotic, respectively. Eleven (52%) children in the azithromycin group and 11 (33%) in the beta-lactam group met the definition for treatment failures (P = 0.34). Eight treatment failures while receiving azithromycin were caused by pneumococci with the macrolide-resistant (M) phenotype, 2 with the macrolide-, lincosamide- and streptogramin B-resistant (MLSB) phenotype and 1 by a macrolide susceptible organism. In the beta-lactam group 7 had a penicillin-resistant isolate, 3 had an intermediately susceptible isolate and 1 had a susceptible isolate. CONCLUSIONS: Our study suggests that treatment failures among patients who developed invasive disease within 30 days of receiving an antimicrobial occur as frequently in patients who receive beta-lactam antibiotics as in those who receive azithromycin. Furthermore macrolide resistant organisms are not more likely to be recovered after a macrolide treatment failure than a penicillin nonsusceptible isolate being recovered after a beta-lactam treatment failure (P = 1.0). PMID- 15131462 TI - Relationship among peripheral leukocyte counts, etiologic agents and clinical manifestations in acute otitis media. AB - OBJECTIVES: To analyze the peripheral leukocyte counts of children with acute otitis media (AOM) in relation to etiology, age, clinical symptoms and signs, prior antibiotic treatment, previous AOM history and pathogen eradication during antibiotic therapy. PATIENTS AND METHODS: Leukocyte counts were determined at diagnosis and on days 4 to 6 of therapy in patients age 3 to 36 months with AOM enrolled in double tympanocentesis studies. Clinical status was determined by a clinical score evaluating severity of fever, irritability and tympanic membrane redness and bulging. RESULTS: Of 771 enrolled patients, culture-positive middle ear fluid was reported in 590 (77%): 294 (50%) Haemophilus influenzae; 150 (25%) Streptococcus pneumoniae; 127 (21%) H. influenzae and S. pneumoniae together; 9 (2%) Moraxella catarrhalis; and 10 (2%) others. Mean leukocyte count +/- sd in patients with AOM caused by S. pneumoniae (15.7 +/- 6.7 cells x 10/mm) was significantly higher than those of patients with AOM caused by H. influenzae (13.7 +/- 5.8 cells x 10/mm) and patients with culture-negative AOM (13.8 +/- 5.6 cells x 10/mm), P < 0.01 for each comparison. Mean absolute neutrophil count (ANC) +/- sd was higher in patients with AOM caused by S. pneumoniae (8.6 +/- 5.1 cells x 10/mm) than in patients with AOM caused by H. influenzae (6.5 +/- 4.0 cells x 10/mm) or culture-negative patients (6.5 +/- 4.4 cells x 10/mm), P < 0.05 for each comparison. When multivariant regression analysis was used, S. pneumoniae etiology and fever (body temperature > or = 38 degrees Celsius) were independent factors, each significantly associated with leukocyte and ANCs. A significant correlation was found between leukocyte and ANCs and higher clinical scores in patients with pneumococcal AOM (P = 0.01, r = 0.21 and P < 0.01, r = 0.27, respectively). The mean leukocyte count on Days 4 to 6 was lower than on Day 1 (11.5 +/- 4.1 compared with 14.2 +/- 6.0 cells x 10/mm, P < 0.01). When paired (Day 1 and Days 4 to 6) examinations were analyzed, the leukocyte counts were lower on Days 4 to 6 irrespective of bacterial eradication or persistence. CONCLUSIONS: Significantly higher leukocyte counts and ANCs were found in pneumococcal AOM than in AOM caused by H. influenzae or in culture-negative AOM. A significant decrease in leukocyte counts was found during antibiotic therapy for AOM, regardless of etiology and bacteriologic outcome. Isolation of S. pneumoniae and fever were each significantly associated with increased peripheral leukocyte and ANCs. PMID- 15131463 TI - Impact of wheezing after respiratory syncytial virus infection on health-related quality of life. AB - BACKGROUND: Respiratory syncytial virus (RSV) lower respiratory tract infection (LRTI) is often followed by recurrent wheezing episodes during childhood. The effect of postbronchiolitis wheezing on the well-being of the child is not known. This study aimed to determine the impact of RSV LRTI hospitalization before age 13 months on health-related quality of life (HRQoL) at age 3 years. METHODS: HRQoL was measured in a previously described cohort of children with a history of RSV infection and compared with control term children from the open population. HRQoL was determined during the winter season in index and control children. We used a validated questionnaire, which measures the health status of children in 13 domains weighted by the impact of the health status problems on well-being. The lung domain was the focus of this study. In addition daily respiratory symptoms of the index children were recorded prospectively up to age 3 years and correlated with HRQoL scores. RESULTS: HRQoL was lower in index children (n = 128) than in control children (n = 340) for the lung, gastrointestinal tract and sleeping domains. HRQoL scores for social and developmental domains were similar for index and control children. As anticipated the largest difference between index and control children was found in the lung domain (77.6 +/- 2.2 vs. 93.9 +/ 0.8, P < 0.01). HRQoL in the lung domain was lower during the winter than during the summer season (77.6 +/- 2.2 vs. 85.6 +/- 1.9, P < 0.01), which was explained by increased airway morbidity during the winter season. We found a high correlation between lung HRQoL scores and total number of wheezing days during follow-up. CONCLUSIONS: Postbronchiolitis wheezing has broad implications for long-term well-being of children. Decreased HRQoL was attributed to postbronchiolitis wheezing, but not to preexistent risk factors, such as premature birth. This study underscores the importance of developing new strategies to prevent and treat long term airway morbidity after RSV LRTI. PMID- 15131464 TI - Driscoll Children's Hospital respiratory syncytial virus database: risk factors, treatment and hospital course in 3308 infants and young children, 1991 to 2002. AB - BACKGROUND: Treatment of respiratory syncytial virus (RSV) lower respiratory tract infection has historically been one of the most frequent reasons for admission to Driscoll Children's Hospital. OBJECTIVE: The objective of this study was to examine the relationship of risk factors for a severe and complicated disease course to the treatment and hospital length of stay. METHODS: Subjects were identified through a retrospective review of the medical records of all patients discharged with a diagnosis of RSV lower respiratory tract infection during 9 of the 11 RSV seasons between July 1, 1991 and June 30, 2002. The RSV seasons from 1991-1992 to 1994-1995 were compared with the RSV seasons from 1995 1996 to 2001-2002 with regard to treatment and hospital course. RESULTS: There were a total of 3308 admissions. Compared with patients with no risk factors, higher percentages of patients with age <6 weeks, history of prematurity, congenital heart disease and neurologic disease were admitted to the pediatric intensive care unit (PICU) and required mechanical ventilation (P < 0.001). Also the hospital length of stay was longer for patients with each of these individual risk factors (P < 0.001). The hospital length of stay and the percentages of patients admitted to the PICU and requiring on mechanical ventilation increased as the number of risk factors increased from zero to 3 or more (P < 0.001). Of patients with 3 or more risk factors, the average hospital length of stay was 13.5 days; 67% were admitted to the PICU, and 47% required mechanical ventilation. Ribavirin use decreased in patients with each of the individual risk factors (P < 0.001) as well as in patients with one or more risk factors (P < 0.001). At the same time the PICU admission rate increased from 6.1% to 11.2% (P < 0.001). CONCLUSIONS: Patients with three or more risk factors were at very high risk for having a severe or complicated disease course associated with admission to the PICU, placement on mechanical ventilation and a longer hospital length of stay. PMID- 15131465 TI - Tumor necrosis factor alpha -- 308 polymorphism associated with increased sepsis mortality in ventilated very low birth weight infants. AB - BACKGROUND: Sepsis commonly complicates the clinical course of critically ill very low birth weight infants, with as many as 30% developing hospital-acquired bacteremia. The tumor necrosis factor alpha (TNF-alpha) -- 308 G/A single nucleotide polymorphism (SNP) is associated with adverse outcome in septic adult patients. METHODS: One hundred seventy-three mechanically ventilated very low birth weight infants were genotyped for the TNF-alpha -- 308 G/A SNP. RESULTS: One hundred twenty (69%) infants were homozygous GG, 45 (26%) were heterozygous AG and 8 (5%) were homozygous AA; 2 of 120 (2%) infants developed early bacteremia in the GG group, and 1 of 53 (2%) developed early bacteremia in the AA/AG group (P = 0.919). One or more episodes of late bacteremia/fungemia developed in 59 of 120 (49%) infants with the GG genotype and 23 of 53 (43%) infants with the AG/AA genotype (P = 0.484). Endotracheal tube colonization rates were 65 of 120 (54%) for infants with the GG genotypes and 28 of 53 (53%) for infants with the AG/AA genotypes (P = 0.871). Nosocomial pneumonia developed in a similar number of infants in both genotype groups (9 of 120 infants vs. 3 of 53 infants; P = 0.461). Mortality from sepsis was 3 times greater in infants with the AA/AG genotypes than in those with the GG genotype (10%vs. 3%; P = 0.038). This difference in sepsis mortality was even greater when only bacteremic/fungemic infants are considered (4 of 59 infants vs. 6 of 23 infants; P = 0.026). CONCLUSIONS: These data suggest that the TNF-alpha -- 308 A allele does not affect the development of sepsis in ventilated premature infants but may increase mortality once sepsis develops. PMID- 15131466 TI - Dosage escalation, safety and immunogenicity study of four dosages of a tetravalent meninogococcal polysaccharide diphtheria toxoid conjugate vaccine in infants. AB - BACKGROUND: Young children have the highest incidence of meningococcal infection. Approximately 50% of disease in United States children less than 2 years of age is caused by serogroups C and Y. In the developing world, serogroups A and W-135 cause outbreaks and epidemics of infection. METHODS: Three groups of 30 infants were enrolled. The first group of infants was given 3 doses of a quadrivalent (group A, C, Y, W-135) polysaccharide meningococcal vaccine conjugated to diphtheria toxoid (MCV-4) at a dosage of 1 microg of each serogroup polysaccharide. The second group of infants was given MCV-4 at a dosage of 4 microg, and the last group of children received a 10-microg dosage. Vaccinations were given at 2, 4 and 6 months of age.A subset of these children was vaccinated at 15 to 18 months of age with licensed meningococcal polysaccharide (A, C, Y, W 135) vaccine. Serum bactericidal antibody (SBA) titers were measured with baby rabbit complement. RESULTS: The proportion of infants with local reactions increased significantly with increasing dosages after Injection 1 and 3. Approximately 1 month after completion of the primary series, the proportion of infants with an SBA titer > or = 1/8 ranged from 54 to 92%, depending on the serogroup and dose of polysaccharide contained in the vaccine. The SBA geometric mean titer varied from 17.4 to 101.6. There was no statistically significant difference between the SBA responses among the 3 dosage groups. After vaccination with polysaccharide vaccine at 15 to 18 months of age, mean fold increases in SBA of 4.9 to 170.3 were observed, suggesting an anamnestic response. CONCLUSIONS: MCV-4 appears to have a reactogenicity profile acceptable to parents and health care providers. It was only modestly immunogenic in infants, but it appeared to prime the immune system of the majority of infants given three doses in infancy. There is no statistically significant immunologic advantage conferred by increasing the dosage beyond 4 microg/ml, and local reactions are more frequent after the 10-microg/ml dosage. PMID- 15131467 TI - Outbreak of human metapneumovirus infection in norwegian children. AB - INTRODUCTION: Human metapneumovirus (hMPV) was recently discovered in children with acute respiratory tract infection. We have studied the occurrence of hMPV and report clinical findings of 50 hMPV-infected children who were hospitalized during an outbreak in Norway. METHODS AND POPULATION: During 5 months from November 15, 2002 to April 14, 2003 we collected nasopharyngeal aspirate specimens from 236 children admitted because of respiratory tract infection (RTI). Samples were analyzed for influenza virus A/B, parainfluenza viruses 1, 2 and 3 and respiratory syncytial virus by direct immunofluorescence assays and cell culture. Rhinovirus, adenovirus and hMPV were identified by polymerase chain reaction. RESULTS: Human metapneumovirus was identified in 50 of 236 children (21%). Most (41 of 50) hMPV-infected children were hospitalized between November 15 and January 15, and during these 2 months hMPV was the most common isolate (41 of 72 isolates; 57%). Respiratory syncytial virus was identified in 36 children (15%), among whom 34 were admitted after the hMPV outbreak. The median age of hMPV-infected children was 12 months (range, 1 to 115 months), and one-half of the children had an underlying chronic disease. The most common symptoms were fever (86%), cough (90%), dyspnea (80%), wheezing (56%), rhinorrhea (44%), anorexia (48%) and vomiting (36%). Eight (16%) had an upper respiratory tract infection (rhinopharyngitis, n = 6; laryngitis, n = 2), 24 (48%) had bronchiolitis and 17 (34%) had pneumonia. Two-thirds with a lower RTI also had signs of upper RTI. Fourteen (28%) children needed supplemental oxygen, 1 was treated with continuous positive airway pressure and 2 were ventilated mechanically. CONCLUSION: Human metapneumovirus was the most common virus isolate during the winter season 2002 to 2003 in children hospitalized for respiratory tract infection. Upper respiratory tract infections and mild to severe bronchiolitis were most common, but a relatively high proportion of hospitalized children developed severe pneumonia. PMID- 15131468 TI - Predictors of virologic response to Lamivudine treatment in children with chronic hepatitis B infection. AB - BACKGROUND: Some children with chronic hepatitis B develop advanced liver disease. Lamivudine, an oral nucleoside, is a therapeutic option. A recent large, multicenter study demonstrated that lamivudine was superior to placebo in eliciting loss of hepatitis B e antigen (HBeAg) and hepatitis B virus (HBV) DNA from serum in children (2 to 17 years) treated for 52 weeks. OBJECTIVE: To identify pretreatment factors that predict the likelihood of response to lamivudine in children with chronic hepatitis B infection. STUDY DESIGN: Data from the multicenter trial in 297 children (191 lamivudine, 96 placebo) were analyzed for the effects of baseline factors on the likelihood of responses. These responses included virologic response, defined as loss of HBeAg and HBV DNA, and HBeAg seroconversion, defined as loss of HBeAg and development of antibody to HBeAg. Univariate and multivariate analyses examined the effects of lamivudine treatment, age, gender, race, body weight, body mass index, previous interferon treatment and baseline alanine aminotransferase (ALT), histologic activity index (HAI) score and HBV DNA on the virologic responses. RESULTS: In the univariate analysis higher baseline ALT, higher HAI score and lower HBV DNA level predicted a greater likelihood of virologic responses to lamivudine. In the multivariate model only baseline ALT and HAI score were predictive of responses. There was no effect of age or ethnicity on response. CONCLUSIONS: Children with higher pretreatment ALT and HAI scores are most likely to respond to lamivudine. Age, ethnicity and other factors do not significantly influence the frequency of virologic responses in children with chronic hepatitis B infection. PMID- 15131469 TI - Immunologic changes during unplanned treatment interruptions of highly active antiretroviral therapy in children with human immunodeficiency virus type 1 infection. AB - BACKGROUND: There are few data on the decline of CD4 cells or percentage during unplanned treatment interruptions in HIV-infected children. METHODS: Data were analyzed on children undergoing interruptions of three or four drug antiretroviral therapy (HAART) in the United Kingdom and Irish Collaborative HIV Paediatric Study and from Sophia Children's Hospital, Rotterdam. Children were included if they had taken HAART continuously for 3 months or longer before interruption and stopped for at least 4 weeks. The effects of CD4 at treatment initiation and of age, CD4, HIV RNA and disease stage at interruption on the CD4 decline during interruption were explored with linear mixed models. RESULTS: Seventy-one children (median age, 7.0 years) had experienced 82 unplanned interruptions of median duration 4.1 (interquartile range, 2.3 to 7.6) months. HAART was restarted after 59 (72%) interruptions (17 with the same and 42 with a new regimen). In children restarting HAART, median CD4% increased toward preinterruption levels by 6 months, when 46% had HIV RNA <400 copies/ml (vs. 15% at interruption). The rate of CD4% decline in 51 children with a median of 3 (range, 2 to 10) CD4 measurements was 0.52% per month (6.2% annually); it was independent of age, CD4 at HAART, preinterruption CD4 or HIV RNA or previous AIDS. CONCLUSIONS: Although the average CD4 decline after stopping HAART appears similar to that in adults, large variability suggests that when designing pediatric trials of planned treatment interruptions, interruption length could be determined by time taken for CD4 to decline below a threshold, rather than by imposing interruptions of fixed duration. PMID- 15131470 TI - Herpes zoster in otherwise healthy children. AB - In normal infants and children, zoster can occur at any time after varicella or varicella vaccination. It is usually diagnosed clinically: a unilateral vesicular eruption following a dermatome or dermatomes. The incidence of zoster increases with age, although children who have had varicella during the first year of life (or in utero) are at increased risk of developing zoster. The incidence of zoster is less after varicella vaccination than after natural infection. Zoster in children is frequently mild, postzoster neuralgia rarely if ever occurs, and antiviral therapy is usually not needed. In a previously normal child with zoster, if the history and physical examination are normal, a laboratory search for occult immunodeficiency or malignancy is not needed. We present five cases of zoster in healthy children and review zoster in the pediatric age group. PMID- 15131472 TI - Rationale and prospects for a nontypable Haemophilus influenzae vaccine. PMID- 15131473 TI - Refractory Kawasaki disease. PMID- 15131474 TI - No evidence of Mycoplasma pneumoniae in acute myringitis. AB - Our aim was to discover Mycoplasma pneumoniae in bullous and hemorrhagic myringitis in children <2 years of age. Middle ear fluid samples (n = 37) and samples taken from the blisters of the tympanic membranes (n = 12) studied by polymerase chain reaction for M. pneumoniae were negative. This study does not support an important role for M. pneumoniae as an etiologic agent in acute myringitis. PMID- 15131475 TI - Interferon-alpha and lamivudine combination therapy of children with chronic hepatitis b infection who were interferon-alpha nonresponders. AB - Greater than one-half of children with chronic hepatitis B infection are nonresponders to interferon-alpha (IFN-alpha). The aim of this study was to investigate the efficacy of lamivudine (LMV) and IFN-alpha combination therapy in these children. Nineteen children were given LMV alone for 3 months; then IFN alpha was added to LMV for 6 months. Virologic response was achieved in seven (36.8%) patients. LMV and IFN-alpha combination therapy may represent an effective treatment option. PMID- 15131476 TI - beta-Hemolytic group F streptococcal bacteremia in children. AB - Group F beta-hemolytic streptococci cause purulent disease and bacteremia in adults. Infections with these organisms are rare in previously healthy children. We report three cases of group F beta-hemolytic streptococcal bacteremia in previously healthy infants and children diagnosed at a single institution during a period of 10 years. Two patients had associated meningitis, and one patient had perforated appendicitis with peritonitis. PMID- 15131477 TI - Successful treatment of a primary cutaneous zygomycosis caused by Absidia corymbifera in a premature newborn. AB - Mucormycosis (zygomycosis) normally occurs among individuals with predisposing factors such as prematurity, use of broad spectrum antibiotics, metabolic acidosis or advanced stages of immunosuppression. There have been reports of sporadic cases of cutaneous mucormycosis related to predisposing skin lesions and contact with contaminated material such as adhesive bandages and tongue depressors placed close to intravenous catheter insertion sites. We report successful treatment of a case of Absidia corymbifera infection with the combination of amphotericin B and surgical debridement of the affected area. PMID- 15131478 TI - Late onset Candida parapsilosis endocarditis after surviving nosocomial candidemia in an infant with structural heart disease. AB - The incidence of nosocomial candidemia is increasing. Late onset endocarditis after surviving nosocomial fungemia is described in adults. We report a case of late onset Candida parapsilosis endocarditis of a structurally abnormal aortic valve in an infant surviving nosocomial candidemia. PMID- 15131479 TI - Otitis media associated with Vibrio alginolyticus in a child with pressure equalizing tubes. AB - Vibrio alginolyticus is an unusual cause of otitis media. Infection usually occurs in the presence of a chronically perforated eardrum or patent pressure equalizing tube. Infection with V. alginolyticus can occur after even mild, brief exposure to seawater, and the interval between exposure to seawater and onset of clinical infection can be prolonged. PMID- 15131480 TI - Resistant Mycobacterium bovis Bacillus Calmette-Guerin disease: implications for management of Bacillus Calmette-Guerin Disease in human immunodeficiency virus infected children. AB - Guidelines for the diagnosis and management of Bacillus Calmette-Guerin (BCG) disease in children are lacking, and there are limited data on drug resistance of Mycobacterium bovis BCG. A 6-month-old HIV-infected infant presented with right axillary adenitis ipsilateral to the site of BCG immunization. M. tuberculosis complex was cultured from axillary lymph nodes and gastric aspirates, and M. bovis BCG was isolated. Susceptibility testing before initiation of therapy demonstrated inherent resistance to isoniazid. The organism acquired rifampin resistance during therapy. This was confirmed by the presence of a mutation in codon 531 (Ser531Tyr) of the rpoB gene. Treatment guidelines for BCG disease with consideration of inherent and possible acquired drug resistance should be established in settings with high rates of vertical HIV transmission and routine BCG vaccination. PMID- 15131481 TI - Complex febrile seizures associated with influenza A. PMID- 15131482 TI - Aseptic meningitis and urinary infection. PMID- 15131484 TI - Retrospective diagnosis of congenital cytomegalovirus infection using dried umbilical cords. PMID- 15131485 TI - Monkeypox and medical ethics. PMID- 15131487 TI - Single-isomer science: the phenomenon and its terminology. AB - Single-isomer drugs are of great importance in modern therapeutics. In this article, the basics of the underlying phenomenon are explained. Some molecules are chiral, ie, their mirror image is not superposable on the original. The most common element producing molecular chirality is a chiral center, typically a carbon atom carrying four different groups. The mirror-image molecules are termed enantiomers, but the less specific terms stereoisomers and isomers are also used. A substance consisting of only one of the two enantiomers is a single enantiomer or single isomer, and the 1:1 mixture of the enantiomers is the racemic mixture or racemate. A graphical convention that conveys the three-dimensional aspects of chiral molecules drawn in two dimensions, as well as two nongraphical conventions, based on optical rotation and configuration, are used to identify enantiomers. Optical rotation is a physical property of single enantiomers and involves rotation of the plane of plane-polarized light, each pure enantiomer rotating with equal magnitude but in the opposite direction (dextro and levo). Configuration is the actual arrangement in space of the atoms of chiral molecules. Two systems of indicating configuration are in use. One employs D and L to denote the respective enantiomers, and is applicable only to a-amino acids and carbohydrates. The other is a universal system using R and S as descriptors for the two possible arrangements, respectively, of the atoms around the chiral center. Interest in chiral drugs stems from the frequently observed biological differences between enantiomers. Such enantioselectivity is the result of different interactions of the drug enantiomers with target receptors that are themselves chiral and single-enantiomeric. PMID- 15131488 TI - The development of single-isomer molecules: why and how. AB - Until relatively recently the three-dimensional nature of drug molecules has been largely neglected, with approximately 25% of marketed drugs being mixtures of agents rather than single chemical entities. These mixtures are not combinations of drugs but mixtures of stereoisomers, generally racemates of synthetic chiral drugs. The individual enantiomers present in such mixtures frequently differ in both their pharmacodynamic and pharmacokinetic profiles as a result of stereochemical discrimination on interaction with chiral biological macromolecules (enzymes and receptors). The use of such mixtures may present problems if their adverse effects are associated with the less active stereoisomer or do not show stereoselectivity. In addition, interactions between enantiomers may occur such that the observed activity of the racemate is not simply the product of the effects of the individual enantiomers. Since the mid 1980s there has been an ongoing "racemate-versus-enantiomer" debate with the potential advantages of single-isomer products, including improved selectivity of action and potential increase in therapeutic index, being highlighted. As a result, regulatory authorities have issued guidelines for dealing with chiral molecules, and the number of single enantiomer agents presented for evaluation has increased. Racemic mixtures may still be developed but require justification such that the risk-benefit ratio may be assessed. In addition to new chemical entities, a number of "old" mixtures are being re-examined as potential single isomer products, the chiral switches, with the potential for an improved therapeutic profile and possibly new indications. However, for the majority of agents currently marketed as mixtures, relatively little is known concerning the pharmacological or toxicological properties of the individual enantiomers. PMID- 15131489 TI - Single isomer versus racemate: is there a difference? Clinical comparisons in allergy and gastroenterology. AB - Many commonly prescribed drugs exist as a mixture of two distinct chiral isomer forms (enantiomers), each with its own unique chemistry, receptor affinity, and pharmacokinetic profile. Much is unknown concerning the clinical utility of these single enantiomers. This review of the stereoisomers of two commonly used drugs- albuterol for asthma and omeprazole for gastroesophageal reflux disease (GERD) and peptic ulcers--examines the improved efficacy, pharmacokinetics, decreased adverse effects, and fewer drug-drug interactions associated with single enantiomers. PMID- 15131490 TI - Great expectations in stereochemistry: focus on antidepressants. AB - Chirality has become an increasingly important consideration in the development of psychoactive drugs because enantiomers often show major differences in their pharmacokinetic and pharmacologic properties. This review illustrates the implications of stereochemistry in clinical psychopharmacology using the antidepressant class of drugs as a focus. In many cases, a better understanding of stereochemistry can improve therapeutic outcomes. For example, with citalopram, the racemic formulation is effective for depression as well as panic and obsessive-compulsive disorders. However, the S-enantiomer, escitalopram, is at least twice as potent as racemic citalopram as an inhibitor of serotonin reuptake, implying that it can be used at lower doses, while offering an improved therapeutic index as well as an improved safety profile and reduced drug interaction liability. Clinical trial data support these advantages. Continuing research on the stereochemical properties of psychoactive drugs should simplify the characterization of dose-response relationships, and clarify the effects of disease states, genetic polymorphisms, pregnancy, age, and gender on stereoselective pharmacokinetics and pharmacodynamics. Better understanding of the fate of chiral psychotropic agents and the factors that influence their stereoselective disposition and actions will provide a rational basis for their expanded use in various patient populations. PMID- 15131491 TI - Escitalopram: a second-generation SSRI. AB - Serotonin (5-hydroxytryptamine, 5-HT) has long been suspected to play a role in the etiology of depression, and modern neurochemical techniques have confirmed this suspicion. Furthermore, all drugs known to be selective (a relative term) serotonin transporter (SERT) inhibitors are effective antidepressants. Of the selective serotonin reuptake inhibitors (SSRIs) approved in a number of countries for use in depression, panic disorder, and obsessive-compulsive disorder, citalopram is the most selective. Citalopram has been used worldwide to treat an estimated 35 million patients, with an excellent safety record. Citalopram is a racemic drug, and its effects on serotonin transport are thought to reside in the S-enantiomer, known as (S)-citalopram or escitalopram. Escitalopram is the most selective SSRI yet developed. Its receptor binding properties and activity in preclinical animal models of depression predict that escitalopram would be effective in the treatment of depression, with approximately twice the potency of the racemate. The pivotal clinical trials of escitalopram not only support this conclusion, but also suggest escitalopram possesses advantages over citalopram in terms of both efficacy and safety. In conclusion, escitalopram is a promising candidate for use as a first-line antidepressant. PMID- 15131492 TI - Efficacy comparison of escitalopram and citalopram in the treatment of major depressive disorder: pooled analysis of placebo-controlled trials. AB - BACKGROUND: Citalopram is a racemic selective serotonin reuptake inhibitor (SSRI) indicated for the treatment of depression. Citalopram is a racemate and its serotonin reuptake inhibitory activity resides primarily in the single S-isomer, escitalopram, which is now being evaluated for its potential usefulness in the treatment of depression and other psychiatric disorders. RESULTS from placebo controlled studies that also included citalopram as an active control have shown that escitalopram is effective in treating depression and associated symptoms of anxiety. However, none of these studies was powered sufficiently to detect differences between active treatment groups. The goal of the present analysis is to evaluate the efficacy of escitalopram compared with citalopram in the treatment of major depressive disorder. METHOD: Data were pooled from three similarly designed, randomized, double-blind, placebo-controlled trials of escitalopram (10-20 mg/day) and citalopram (20-40 mg/day). Patients were male or female, greater than or equal to 18 years of age, who met criteria for a major depressive episode with a Montgomery Asberg Depression Rating Scale (MADRS) score greater than or equal to 22 at baseline. Efficacy measures included change from baseline in MADRS score and the Clinical Global Impression of Improvement (CGI-I) scale. Improvement in associated symptoms of anxiety was measured using the change from baseline in the MADRS inner tension item. RESULTS: Both escitalopram and citalopram significantly improved depression and anxiety symptoms compared with placebo, and there were significantly more MADRS responders (defined as >/=50% improvement in MADRS scores at end point) in the escitalopram and citalopram treatment groups. Escitalopram treatment was associated with statistically significant improvements in all efficacy measures relative to placebo after 1 week of treatment, whereas citalopram treatment statistically separated from placebo at the end of week 4 (CGI-I and MADRS inner tension) or week 6 (MADRS). Escitalopram treatment also was statistically significantly superior to citalopram treatment at a number of time points. CONCLUSION: These data support the effectiveness of escitalopram and citalopram in the treatment of major depressive disorder, and suggest escitalopram may have a faster onset and greater overall magnitude of effect than citalopram in improving symptoms of depression and anxiety in patients with major depressive disorder. PMID- 15131493 TI - New single-isomer compounds on the horizon. AB - In 1992, the Food and Drug Administration (FDA) issued new guidelines governing stereoisomerism in new-drug development. The guidelines strongly encourage the development of single isomers and discourage stereoisomeric (eg, racemic) mixtures. As a result, most new chiral drugs are being developed as single enantiomers (ie, single isomers). There are three mechanisms for the identification and development of new single-isomer drugs: chiral switches (CS), chiral metashifts (CM), and new single-isomer chemical entities (NSICEs). In a CS, one of the two enantiomers of an established racemate is developed as a new drug, with the expectation that the single-isomer form has advantages over the racemic parent in terms of efficacy and/or adverse effects. Many new CS drugs are in development, eg, (S)-oxybutynin for urinary incontinence and escitalopram for depression. In a CM, a chiral metabolite of a drug is developed, in single-isomer form, as an agent with advantages over the parent. Among the current CM drugs in development are (+)-norcisapride (safer GI prokinetic agent than the racemic parent cisapride) and (S)-desmethylzopiclone (antianxiety agent, metabolite of the sedative-hypnotic zopiclone). Many NSICEs are in development, eg, rosuvastatin as an antihypercholesterolemic, posaconazole as an antifungal, sitafloxacin as a fluoroquinolone antibacterial, pregabalin as an anticonvulsant, abarelix as an antineoplastic, etc. As in the development of any new drug, not every single-isomer candidate will reach the clinic, but there is no doubt that the move to single-isomer agents is an important step forward in the search for better and safer drugs. PMID- 15131494 TI - Conclusion: the future of single-isomer pharmacology. PMID- 15131495 TI - [Subjective evaluation of medical images by using ROC analysis]. PMID- 15131496 TI - [Consider the future radiography of the bones and the joints together]. PMID- 15131497 TI - [Symposium I. Usefulness of cardiac function assessment by gated SPECT. Introduction]. PMID- 15131498 TI - [Comparison of the cardiac functional analysis software in gated SPECT]. PMID- 15131499 TI - [Reliability of gated SPECT and the present conditions of a difference between institutions]. PMID- 15131500 TI - [Comparison of modalities in cardiac function analysis]. PMID- 15131501 TI - [Clinical usefulness of ECG-gated SPECT]. PMID- 15131502 TI - [ A future view of gated SPECT]. PMID- 15131503 TI - [Harmonic imaging]. PMID- 15131504 TI - [The expectation of the doctor acquisition in the radiological technologists]. PMID- 15131505 TI - [Digital mammography: current status and image processing]. PMID- 15131506 TI - [2003 Standardization Committee activities]. PMID- 15131507 TI - [New hope for winning the war against cancer: IMRT=Intensity-Modulated-Radiation Therapy -Part 10-]. PMID- 15131508 TI - [Thyroid uptake error in four different gamma camera systems]. AB - The thyroid uptake of (123)I and (131)I is generally measured by a gamma camera system. We evaluated the error in determining thyroid uptake caused by different methods of calculation among four gamma camera systems with various collimators. We first designed an original thyroid phantom that consisted of the thyroid and a body containing various levels of radioiodine activity. The applications for thyroid uptake equipped in two gamma camera systems performed calculations by the automatic method with background counts not subtracted from the capsule counts. When the size of the rectangular region of interest (ROI) for the capsule was set at 10x8 cm (a typical ROI size for the thyroid), percentages of thyroid uptake as calculated by the manual method with background counts subtracted from the capsule counts and thyroid counts were 52% to 57% when the value was set at 55% for (123)I; and 54.2% and 58.7%, respectively, when the value was set at 60% for (131)I. On the other hand, the percentages of thyroid uptake calculated by the automatic method with the application using two gamma camera systems with non subtraction of background counts from the capsule counts were 46% and 50.5% when the value was set at 55%; and 49.6% when the value was set at 60%. The values calculated by the automatic method were underestimated as a result of background counts that were not subtracted from the capsule counts. When ROI size for the capsule was set at 4x4 cm, which is slightly larger than the capsule size, even thyroid uptake as determined by the automatic method using the application showed a difference of less 2% from the set values. There was no difference in thyroid uptake among the various kinds of collimators, high-resolution collimators, all purpose collimators, and a suitable collimator for gamma-ray energy of (123)I. PMID- 15131509 TI - [Trial of finger contamination reduction of the operator in nerve block treatment: comparison of over- and under-table systems]. AB - Fluoroscopy-guided intervention of the lumbar spine, such as nerve block, plays an important role in the management of disc hernia patients. However, irradiation of operators' fingers remains a problem even with careful collimation and operation, especially when performed by non-radiologists. We compared the irradiation doses of under-table and over-table fluoroscopy systems, and we discuss the most advantageous method of reducing irradiation. The effectiveness and conditions of use of lead protection gloves were also evaluated. Skin dose was monitored using polymethyl methacrylate (PMMA) and an electronic dose meter. The skin doses of over- and under-table fluoroscopy were compared using C-arm fluoroscopy. Finger irradiation dose with 0.03 mmPb protection gloves was also measured. The under-table method reduced skin dose by 95% compared with the over table method. Thicker PMMA resulted in a higher rate of irradiation reduction. Protection gloves reduced radiation dose by half, although this reduction was cancelled when automatic brightness control (ABC) was utilized. Under-tube fluoroscopy was superior to over-tube fluoroscopy in reducing irradiation to the fingers. PMID- 15131510 TI - [Full-field digital mammography with amorphous silicon-based flat- panel detector: physical imaging characteristics and signal detection]. AB - The physical characteristics of a clinical amorphous silicon-based flat-panel imager for full-field digital mammography were investigated. Pre-sampled modulation transfer functions (MTF) were measured by using a slit method. Noise power spectra were determined for different input exposures by fast Fourier transform. The MTFs of full-field digital mammography systems showed significantly higher values than those of the computed radiography (CR) system. The full-field digital mammography system showed a lower noise level than that of the CR system under the same exposure conditions. Contrast detail analysis has been performed to compare the detectability of the full-field digital mammography system with that of the screen-film (Min-R 2000/Min-R 2000) system. The average contrast-detail curves of digital and film images were obtained from the results of observation. Image quality figures (IQF) were also calculated from the individual observer performance tests. The results indicated that the digital contrast-detail curves and IQF, on average, are superior to those of the screen film system. PMID- 15131511 TI - [Study of the normal database that affects 3D-SSP Z-score mapping]. AB - The effects of image reconstruction conditions and devices used on 3D-SSP analysis (Z-score mapping) were investigated. Images of the 3D Hoffman brain phantom were taken with the IRIX device or E.CAM. Using two methods of image reconstruction (FBP and OSEM), several kinds of phantom normal databases (PNDB) and signal data, with or without correction for attenuation, were obtained at varying Butterworth filter cutoff frequencies. The results of 3D-SSP analyses were compared between combinations of PNDB and signal data obtained under the same conditions of image reconstruction and combinations obtained under different conditions. The results of PNDB obtained with different devices were also analyzed. For combinations of PNDB and signal data both obtained with OSEM, the Z score of signals was high. When FBP was used to obtain both PNDB and signal data, the Z-score and signal size tended to become greater and the false-positive rate tended to decrease as the Butterworth filter cutoff frequency became lower. Whether or not correction for attenuation was incorporated did not affect the results of analysis when the same method was used to collect both PNDB and signal data. If the method for image reconstruction differed between PNDB and signal data, the false-positive rate was relatively high. Using a different device to obtain PNDB resulted in a relatively high false-positive rate. We therefore recommend that the conditions for image reconstruction from signal data should be matched to the conditions for image reconstruction from PNDB. It seems impossible to share the same NDB among different devices. PMID- 15131512 TI - [Effect of BOLD on sight stimulation and artifact of CSF: identification of activating signal and CSF artifact]. AB - This study used functional magnetic resonance imaging (fMRI) for brain function evaluation. fMRI data were collected by a block paradigm, and brain function was evaluated. In the block paradigm, the BOLD effect causes a several-second delay in hemodynamics, and analytical processing is done in consideration of this time lag. However, irrelevant artifacts caused by the BOLD effect frequently occur in analytical processing. Therefore, there is a limit to obtaining a sufficient activating reaction in the analytical system, which is normally equipped with MRI. Thus, obtaining the activating reaction is limited because the corresponding misregistration correction, angle correction, time series correction, and so on, are insufficient in an analytical system equipped with MRI. It has become standard to use analytical systems such as SPM. We examined the mean curve of the activating part and the area of artifact. As a result, it was possible to identify the activating signal and the artifact signal, and it became possible to obtain an adequate response from a system that is normally equipped with an activating signal in which artifacts are few. PMID- 15131513 TI - [Evaluation of a contrast examination technique for three-dimensional CT angiography (3DCTA) of the head and craniocervical region]. AB - In three-dimensional CT angiography (3DCTA) studies, we make it a rule to use a CT number monitoring system (SureStart, Toshiba Medical Systems Company) to ensure that contrast-enhanced images are acquired at the optimal timing. However, although SureStart can accurately determine enhancement start timing, it is still possible to inject more contrast medium than necessary because the scan start time is not known with certainty. To address this problem, we conducted investigations to determine the ideal contrast examination technique using SureStart and an injector synchronization system. Our results showed that a CT number of 300 HU in the middle cerebral artery (M1) could be obtained with the injection of contrast medium for a period of 45 s (450 mgI/kg) for the head or for 50 s (450 mgI/kg) for the craniocervical region. This makes it possible to perform contrast studies with greater reproducibility by taking individual variation into consideration. Moreover, it was found that comparable results could be obtained by terminating the injection of contrast medium 15 s before the completion of the study and immediately injecting a physiological saline solution flush, permitting the volume of contrast medium to be further reduced. PMID- 15131514 TI - [Evaluation of radiation therapy for keloid using electron beam]. AB - In radiation therapy for keloid, electron beams are delivered to the skin through a lead shield hollowed into the shape of the keloid. The shape of a postoperative keloid scar is linear, causing the irradiation shield to be long and narrow. This lead shield is put on the surface of the skin. Therefore, it is considered that beam data used in general external irradiation are not applicable to irradiation for keloid. Therefore, we used a water equivalent phantom and measured beam data by using chambers or film dosimeters. Experimental conditions were the same as those of actual radiotherapy for keloid. As a result of this procedure, the radiation technique was optimized. Electron energy and thickness of the bolus, thickness of the lead shield, margins such that the planning target volume would receive the necessary dose, and the method of MU calculation all were determined. It was suggested that these experiments were useful to establish the appropriate technique in irradiation for keloid. PMID- 15131515 TI - Neural circuitry of anxiety: evidence from structural and functional neuroimaging studies. AB - Present understanding of the neural circuitry of anxiety has come from a variety of sources, including animal, clinical, and most recently, neuroimaging studies. Evidence from these sources has converged to form a translational bridge from animal models to human pathophysiology. In particular, the classical fear conditioning paradigm has served as a foundation for this bridge. Proposed models for the neural circuitry of normal anxiety as well as the anxiety disorders are discussed. A brief review of specific findings from neuroimaging studies of posttraumatic stress disorder, specific phobia, social phobia, obsessive compulsive disorder, and generalized anxiety disorder is also provided. PMID- 15131516 TI - Applications of neuroreceptor imaging to psychiatry research. AB - Over the past two decades, there have been significant advances in the ability to study the neurochemistry of the living brain using neuroreceptor radiotracers with Positron Emission Tomography (PET) and Single Photon Emission Computed Tomography (SPECT) imaging modalities. The greater availability of radiotracers for neurotransmitter synthesis/metabolism, enzymes, transporters and receptors, as well as neuromodulators and second messengers has enabled the evaluation of hypotheses regarding neurotransmitter function and regulation that are generated from basic neuroscience studies in animals, and the investigation of the neurochemical substrates of psychiatric disorders and the mechanism of action of psychotropic medications. This review will focus on the status of radiotracer development, on the clinical and methodological considerations regarding neurochemical brain imaging study design and data interpretation. The applications of neurochemical brain imaging methods to the study of specific psychiatric disorders, including schizophrenia, anxiety disorders, depression and Alzheimer's Disease, will be reviewed and potential future directions of research in these areas identified. Finally, the studies of the neurochemical substrates of personality traits will be reviewed. Thus far, fundamental observations have been made with respect to 1). detecting abnormalities in the availability of neurotransmitter transporter and receptor sites in psychiatric patients; 2). evaluating the relationship of these neurochemical measures to symptomatology; and 3). assessing the magnitude of occupancy of the initial target sites of action of psychotropic medication relative to treatment response and drug concentrations. Further advances in instrumentation and radiotracer chemistry will enable investigators to conduct pre-clinical and clinical mechanistic studies focused on other neurotransmitters and neuromodulators. These data will provide important insights into the neurochemical substrates of treatment response variability in psychiatric disorders that will have important implications for the refinement of pharmacotherapy. PMID- 15131517 TI - Bupropion sustained release for panic disorder. AB - Despite anecdotal reports suggesting that bupropion may be effective for panic disorder, both clinical lore and the results of one small controlled study suggest otherwise. There remains a paucity of systematic prospective data addressing this issue. Twenty outpatients meeting criteria for panic disorder with or without agoraphobia were entered in an 8 week, two center open-label flexible dose trial of bupropion SR. Treatment with bupropion SR resulted in a clinically and statistically significant mean reduction of 1 to 2 points in the primary outcome measure, the CGI Severity score, in both the intent to treat (ITT; t=4.36, df=19, p<0.001) and completer samples (t=3.89, df=13, p<0.002). Significant improvement was also noted in both completer and ITT samples for all other panic symptom measures, which included the Panic Disorder Severity Scale (PDSS), number of panic attacks in the past two weeks, and the proportion of time anticipatory anxiety was present. Although results may be influenced by the open nature of this trial, our findings suggest that bupropion SR may be effective for the treatment of panic disorder. Further controlled study of its efficacy for this condition are warranted. PMID- 15131518 TI - Continuation treatment of chronic depression: a comparison of nefazodone, cognitive behavioral analysis system of psychotherapy, and their combination. AB - Little is known about the relative benefits of psychotherapy, medication, and combined treatment as continuation therapies for chronic forms of major depressive disorder (MDD) after a positive response to acute treatment. We hypothesize that combined treatment would demonstrate superior continuation phase outcomes compared to either monotherapy, as evidenced by lower relapse rates and greater rates of improvement from partial to full remission. We report 16-week continuation phase outcomes for 324 patients who had participated in either the acute phase of a randomized multicenter trial of nefazodone, Cognitive Behavioral Analysis System of Psychotherapy (CBASP), or combination therapy (COMB) for chronic forms of MDD. Patients entering the continuation phase had either fully or partially remitted after 12 weeks of acute phase treatment. The primary efficacy measure was the 24-item Hamilton Rating Scale for Depression. For patients in remission at acute phase exit, 73.3% (107/146) maintained their remitted status at endpoint of the continuation phase. Of those having a partial remission at acute phase exit, 52.9% (92/174) achieved full remission by end of continuation. A greater proportion of patients maintained a partial or full remission status on COMB (90%) compared to nefazodone (80%, p=0.011) or to CBASP (82%, p=0.042). These differences reflected greater symptom re-emergence in the partial remission groups on CBASP and nefazodone monotherapy compared to COMB. Continuation treatment assignment was not randomized or blinded. There was no placebo group. Most patients with chronic forms of MDD sustained their acute phase response and more than 50% of partial remitters achieved full remission while continuing treatment with nefazodone, CBASP, or COMB. COMB was associated with less symptom re-emergence during the continuation phase than either monotherapy, particularly for partial remitters. PMID- 15131519 TI - Evidence-based practice: how to evaluate what you read. AB - Systematic reviews and meta-analysis are affected by the studies that are included or excluded. A number of biases are common including excluding unpublished data, non English language publications, and non indexed articles. In addition there can be a differential impact based on the quality and comparability of studies. In this article we review the potential significance of these biases. PMID- 15131520 TI - Cholesterol-lowering medication and relapse of depression. AB - This study sought to examine the effects of taking cholesterol-lowering medication on outcomes of depression among older depressed adults. 243 elderly depressed patients were treated using an antidepressant algorithm by a geriatric psychiatrist who administered the Montgomery-Asberg Depression Rating Scale (MADRS) at clinical visits up to six years. We defined remission and relapse as achieving MADRS scores less than 7 and greater than 15, respectively. There were no between-group differences on time to remission. Among 167 remitted patients with follow-up data, 61.4% on cholesterol-lowering medication, and 39.8% not on cholesterol medications experienced a relapse during the follow-up period (p<0.032), and remitted patients taking cholesterol medications relapsed more quickly than did others in adjusted analyses (hazard ratio=1.791, p<0.018). These results add to the literature linking cholesterol and mood, and they support the "vascular depression" hypothesis in geriatric depression. Future studies of depression outcomes in the elderly will need to include serial examination of cholesterol levels. PMID- 15131521 TI - Tryptophan depletion, serotonin, and depression: where do we stand? AB - Tryptophan depletion is a widely used paradigm to study serotonin system-related mechanisms in the pathophysiology and treatment of depression. There is convincing evidence that tryptophan depletion primarily and selectively affects serotonergic transmission. The behavioral data in healthy controls with and without genetic risk for depression, and in patient populations during the symptomatic phase of depression and when being remitted, suggest a trait abnormality of serotonin function in depression and that antidepressants may compensate for the underlying deficit. Tryptophan depletion may be a useful tool to create more integrative models for the pathophysiology of depression that take into account neurobiological systems beyond monoamines. More recent studies combining tryptophan depletion with genetic variables may provide an important approach for studying gene/environment interactions using candidate genes to define endophenotypes, which ultimately will improve currently used diagnostic categories and help to generate more advanced models to understand the neurobiology of depression. This may lead to the development of truly novel treatment approaches for depression. PMID- 15131522 TI - Risperidone in the management of psychiatric and neurodegenerative disease in the elderly: an update. AB - OBJECTIVE: Neuropsychiatric disorders are common among elderly patients in long term care facilities. Although dementia is most common, schizophrenia and schizoaffective disorder, delusional disorder, bipolar disorder, and Parkinson's disease, as well as other psychiatric and neurodegenerative disorders, can also occur in this population. Our objective was to review the literature pertaining to the safety and efficacy of risperidone in elderly patients, the atypical antipsychotic with the longest history of use in this population. We identified original studies of risperidone in elderly patients through MEDLINE and CURRENT CONTENTS database searches. Additional material was identified via abstracts presented at national and international conferences. Published data from controlled and uncontrolled trials in elderly patients support the efficacy of risperidone in controlling behavioral and psychological symptoms of dementia, as well as treating psychotic symptoms associated with schizophrenia and other disorders with psychotic features. Limited data suggest that risperidone might also be useful for management of patients with delirium and Huntington's disease. Risperidone was well tolerated in the elderly and associated with a low risk of movement disorders and anticholinergic effects. A recent change in the product information described an increased incidence of cerebrovascular events in some elderly and suggests the need for further investigations in this population. There is a substantial published database supporting safe and effective use of risperidone for treatment of psychosis, agitation, and aggression in elderly patients. PMID- 15131523 TI - The role of GABA in the pathophysiology and treatment of anxiety disorders. AB - Mechanisms underlying the pathological characteristics of the various anxiety disorders have yet to be fully elucidated. One of the most widely accepted mediators known to play a central role in the pathophysiology of anxiety disorders is the g-aminobutyric acid (GABA) system. Evidence supporting the role of a dysfunctional GABA system has resulted from clinical experience with the benzodiazepines, as well as subsequent determination of mechanism of action, genetic engineering, and neuroimaging studies of the GABA receptor. The concatenation of results suggests a relative deficiency in GABA neurotransmission, which can be augmented by agents acting on different components of the GABA system. Agents such as the benzodiazepines, neuroactive steroids, and barbiturates act as allosteric modulators of the GABAA receptor; b carboline and the barbiturates function as direct GABA agonists. Valproate, gabapentin, pregabalin, and vigabatrin increase brain GABA levels or neurotransmission at least in part by targeting the metabolic pathways of GABA. Tiagabine selectively increases synaptic GABA availability by blocking the reuptake of GABA via transporter inhibition. Evidence exists, to a greater or lesser extent, that all of these agents possess anxiolytic properties, as would be expected by their mechanisms of action. This article reviews the findings implicating the GABA system in the pathophysiology of anxiety disorders and describes the potential role of agents that modulate GABA neurotransmission in the treatment of these disorders. PMID- 15131524 TI - Angiotensin-converting enzyme inhibitors: a new therapy for atrial fibrillation? PMID- 15131525 TI - Role of stress single-photon emission computed tomography imaging in asymptomatic patients with diabetes. PMID- 15131526 TI - Does stenting save lives? Understanding the strengths and limitations of meta analysis. PMID- 15131527 TI - Endomyocardial biopsy: a procedure in search of an indication. PMID- 15131528 TI - Aortic stenosis and hyperlipidemia: establishing a cause-effect relationship. PMID- 15131529 TI - Clinical impact of stent construction and design in percutaneous coronary intervention. AB - Convincing end point data demonstrating the anatomic and clinical superiority of stent placement compared with balloon angioplasty together with significant improvement in stenting technique and poststent management have resulted in an explosion in stenting procedures and the emergence of more than 40 stent types with disparate designs and material composition in clinical use. Structural nuances in design, composition, and coating of different stent models, however, have been shown to have a major influence on the risk of stent thrombosis, the degree of vessel wall injury, and subsequent intimal proliferation in the experimental model. There is now substantial amount of evidence to indicate that the same relationship between stent structural characteristics and vessel wall outcome holds true in humans. This article provides an up-to-date overview of the clinical impact of stent construction and design, including the clinical performance of drug-eluting stents. PMID- 15131530 TI - Coronary heart disease and lipid-modifying treatment in African American patients. AB - African Americans have the highest overall coronary heart disease (CHD) mortality rate of any ethnic group in the United States. They also exhibit a greater prevalence of a number of individual CHD risk factors, especially hypertension and type 2 diabetes mellitus (a CHD risk equivalent) and greater clustering of risk factors. The African-American population is under-represented in lipid lowering clinical end point trials and remains inadequately treated with lipid lowering therapy in the clinical setting; this latter fact is of particular concern because, in the new National Cholesterol Education Program guidelines, many more black patients with hypercholesterolemia should be receiving more intensive lipid-lowering treatment. A number of steps must be taken to improve prospects of CHD risk reduction through lipid-lowering therapy in African Americans. These include improving the understanding of the relationship of risk factors to disease and improving the understanding of both lipid responses to and clinical benefits of lipid-lowering therapy. In addition, because African Americans have a higher prevalence of several modifiable CHD risk factors, this population should be rigorously targeted for risk-reduction strategies, including screening and treatment for hypertension, type 2 diabetes mellitus, and dyslipidemia. Educational outreach programs can provide a key role in raising community awareness of CHD risk factors and potential treatment options. PMID- 15131531 TI - Cardiovascular effects of raloxifene: the arterial and venous systems. AB - BACKGROUND: Cardiovascular disease caused by atherosclerosis is the largest single killer of women. Prior observational data had suggested that hormone therapy may have cardioprotective effects. METHODS: Data from clinical trials and basic science studies were evaluated to assess the cardiovascular effects of hormone therapy and selective estrogen replacement modulators. RESULTS: Hormone therapy does not appear to lower the risk of cardiovascular events in older postmenopausal women. Selective estrogen receptor modulators (SERMS) have been approved for human use; tamoxifen is used for treatment and prevention of breast cancer and raloxifene is used for the treatment and prevention of osteoporosis. Raloxifene is the only SERM being specifically studied for its effects on coronary heart disease events in a prospective, randomized, controlled trial. CONCLUSIONS: Although raloxifene does increase venous thromboembolic events, there is suggestive data that it may have favorable effects on the arterial systems in women. Only compelling positive data from the Raloxifene Use for The Heart (RUTH) trial will lead to greater use of SERMS to potentially lower the risk of atherosclerotic vascular disease. PMID- 15131532 TI - Amiodarone versus implantable cardioverter defibrillator for asymptomatic nonsustained ventricular tachycardia in nonischemic dilated cardiomyopathy. PMID- 15131533 TI - Fractional flow reserve: critical review of an important physiologic adjunct to angiography. AB - With growing numbers of patients with acute coronary syndromes undergoing catheterization without prior non-invasive imaging and the increasing numbers of patients with angiographic multivessel coronary artery disease, complementary physiologic lesion assessment for directing revascularization is of increasing value. Fractional flow reserve (FFR) has emerged as a simple, reliable, and reproducible physiologic index of lesion severity. In concert with coronary flow reserve, FFR yields significant information about the epicardial artery and the microvascular bed it subtends. This review will critically consider the theory of FFR measurements in the context of recent insights into coronary hemodynamics and how FFR measurements are significantly affected by microvascular disease states such as myocardial infarction, left ventricular hypertrophy, or diabetes mellitus. Clinical studies have demonstrated the usefulness of FFR for the assessment of intermediate lesions in single-vessel, multivessel, and possibly left main disease. In addition, measurement of fractional collateral flow reserve during angioplasty and FFR after stent deployment have prognostic value for major adverse cardiac events after percutaneous intervention. Furthermore, with economic concerns about the appropriate use of drug-eluting stents, FFR assessment can be used to direct percutaneous coronary intervention only to flow limiting lesions and to achieve optimal stent deployment. Therefore, FFR represents a powerful new tool in the cardiologists' arsenal in the cardiac catheterization laboratory. PMID- 15131534 TI - Impaired glucose tolerance and cardiovascular disease: the possible role of post prandial hyperglycemia. AB - There is increasing evidence that the post-prandial state is an important contributing factor in the development of atherosclerosis. In subjects with impaired glucose tolerance, whereas fasting glycemia is in reference range, the post-prandial phase is characterized by a rapid and large increase in blood glucose levels. The possibility that this post-prandial "hyperglycemic spike" may be relevant to the development of cardiovascular disease in these subjects has received recently much attention. The oral glucose tolerance test, although highly non-physiological, has been used largely as model of the post-prandial state, and epidemiological studies have shown that impaired oral glucose tolerance is associated with an increased risk of cardiovascular disease, because the glycemia level after 2 hours of the glucose challenge is a direct and independent risk factor. Most of the cardiovascular risk factors are modified in the post-prandial phase and are directly affected by an acute increase of glycemia. The mechanisms through which acute hyperglycemia exerts its effects may be identified in the production of free radicals, which favours the development of an endothelial dysfunction, a prothrombotic and proinflammatory condition. Future studies may evaluate whether correcting the post-prandial hyperglycemia in the impaired glucose tolerance state can form part of the strategy for the prevention and management of cardiovascular diseases in these subjects. PMID- 15131535 TI - Vascular smooth muscle cells and calcification in atherosclerosis. AB - Vascular calcification is a prominent feature of atherosclerosis but the mechanisms underlying vascular calcification are still obscure. Since bone associated proteins such as osteonectin, osteocalcin, and matrix Gla protein have been detected in calcified vascular tissues, calcification has been considered to be an organized, regulated process similar to mineralization in bone tissue. Vascular smooth muscle cells (VSMCs) are currently considered to be responsible for the formation of vascular calcifications. Apoptosis of VSMCs appears to be a key factor in this process, while other factors including cell-cell interactions (macrophages and VSMCs), lipids, and plasma inorganic phosphate levels modulate the calcification process. The focus of this review is on the role of VSMCs in the development of calcifications in atherosclerotic plaques. PMID- 15131536 TI - Impact of coronary artery stents on mortality and nonfatal myocardial infarction: meta-analysis of randomized trials comparing a strategy of routine stenting with that of balloon angioplasty. AB - BACKGROUND: A strategy of routine stenting has been shown to reduce the need for target-vessel revascularization compared with a strategy of balloon angioplasty alone; however, the impact on mortality and frequency of nonfatal myocardial infarction is unclear. The aim of this study was to provide a quantitative comparison of the impact of coronary stenting on the rates of mortality and myocardial infarction with that of balloon angioplasty with provisional stenting. METHODS: We performed a meta-analysis of randomized trials comparing routine coronary stenting to percutaneous transluminal coronary angioplasty (PTCA), including only those trials that used combination antiplatelet therapy (aspirin and a thienopyridine) as an adjuvant to stenting. Such trials included: the Belegian Netherlands Stent Study (BENESTENT) II, Optimal Coronary Balloon Angioplasty With Provisional Stenting Versus Primary Stent (OCBAS), Balloon Optimization vs Stent Study (BOSS), Evaluation of Platelet IIb/IIIa Inhibitor for Stenting (EPISTENT), Optimum Percutaneous Transluminal Coronary Angioplasty Compared With Routine Stent Strategy (OPUS-1), French Optimal Stenting Trial (FROST), Angioplasty or Stent (AS), and Doppler Endpoint Stenting International Investigation (DESTINI) trials for de novo coronary artery lesions; the Stent vs Percutaneous Angioplasty in Chronic Total Occlusion (SPACTO), Total Occlusion Study of Canada (TOSCA), Stent or Angioplasty after Recanalization of Chronic Coronary Occlusions (SARECCO), and Mayo-Japan Investigation for Chronic Total Occlusion (MAJIC) trials for coronary occlusions; the Primary Angioplasty Versus Stent Implantation in Acute Myocardial Infarction (PASTA), Gianturco-Roubin in Acute Myocardial Infarction (GRAMI), Florence Randomized Elective Stenting in Acute Coronary Occlusions (FRESCO), Immediate Coronary Angioplasty with Elective Wiktor Stent Implantation Compared with Conventional Balloon Angioplasty in Acute Myocardial Infarction (STENTUIM-2), Stent Primary Angioplasty in MI (Stent-PAMI), Zwolle, and Controlled Abciximab and Device Investigation to Lower Late Angioplasty Complications (CADILLAC) trials for acute ST-segment elevation myocardial infarction; and the Intracoronary Stenting or Angioplasty for Restenosis Reduction in Small Arteries (ISAR-SMART), Park, Stenting in Small Arteries (SISA), and Bestent in Small Arteries (BESMART) trials for small vessels. RESULTS: The 23 trials enrolled 10,347 patients, with 5130 patients randomized to receive stent and 5217 patients randomized to receive balloon angioplasty. A total of 902 (17 %) of patients crossed over from a strategy of balloon angioplasty to stent placement because of the inability to achieve a satisfactory result with a balloon. No significant difference was observed between the stent group and PTCA group in the rates of death or myocardial infarction, despite a significant reduction in the frequency of major adverse cardiac events (odds ratio, 0.59; 95% CI, 0.50-0.70; P <.001), which was driven entirely by a reduction in target vessel revascularization. CONCLUSIONS: An initial strategy of stent placement versus balloon angioplasty with provisional stenting is associated with a similar mortality rate and frequency of nonfatal myocardial infarction after a mean follow-up period of 12.8 months. Patients who underwent stent placement had a significantly lower risk of major adverse cardiac events only when target revascularization is included as an end point. PMID- 15131537 TI - Angiotensin-converting enzyme inhibitors as adjunctive therapy in patients with persistent atrial fibrillation. AB - OBJECTIVES: The purpose of the current study was to assess the effect of angiotensin-converting enzyme inhibitor (ACEI) therapy in facilitating cardioversion from persistent atrial fibrillation (AF) and maintaining sinus rhythm. BACKGROUND: Pharmacologic therapy and electrical cardioversion for AF are often unsuccessful in maintaining long-term sinus rhythm. METHODS: The current study, a 1-year, prospective follow-up, comprised 47 patients with persistent AF undergoing electrical cardioversion. Patients receiving ACEI were compared with those receiving other medications. The study end point was the number of defibrillation attempts required for atrial defibrillation and the number of hospital admissions. A secondary end point was change in signal-averaged P-wave duration (SAPD) 1 year after successful electrical cardioversion. RESULTS: Of those admitted and requiring electrical defibrillation, the number of defibrillation attempts required for successful cardioversion was significantly less in the ACEI group (P <.001). The incidence rate ratio for admissions comparing recipients of ACEI with others was 0.14 (P =.03). Patients receiving ACEI therapy had significantly lower SAPD at 1 year when compared with the no ACEI group (135 ms +/- 3 vs 150 ms +/- 2, P =.002). CONCLUSIONS: The use of long term ACEI therapy facilitated electrical defibrillation in patients with persistent AF. ACEI therapy also reduced SAPD, suggesting amelioration of the arrhythmogenic substrate. Furthermore, we confirmed that SAPD is prolonged in patients with persistent AF. PMID- 15131538 TI - Non-ST-elevation acute coronary syndrome in the elderly: treatment strategies and 30-day outcome. AB - BACKGROUND: The purpose of this study was to assess the current care of elderly patients with non-ST-elevation acute coronary syndrome (ACS), with particular regard to the rate of use of antiplatelet drugs and the type of strategy, aggressive or conservative, in a population of consecutive patients admitted to 76 Coronary Care Units in Italy. METHODS: Prospective registry of patients admitted to Coronary Care Units with a diagnosis of non-ST-elevation ACS during a 2-month period. Thirty-day follow-up was available in all patients. RESULTS: Of 1581 patients enrolled in the registry, 564 were 75 years or older. As compared with the 1017 younger patients, elderly patients had a greater prevalence of female sex (42% vs 27%, P <.001), hypertension (70% vs 59%, P <.001), prior myocardial infarction (MI) (41% vs 29%, P <.001), prior angina (18% vs 13%, P <.01), prior use of aspirin (49% vs 39%, P <.001), ST-segment depression (54% vs 43%, P <.001), and troponin positivity (66% vs 59%, P <.05). The higher-risk profile of elderly patients was confirmed by the greater number of patients with a high TIMI risk score (37% vs 22%, P <.001). GPIIb/IIIa inhibitors were less frequently used in elderly patients (P <.05). An aggressive strategy (coronary arteriography within 4 days of admission, followed by revascularization, if feasible) was adopted in 39% elderly patients and in 56% younger patients (P <.001). An interventional procedure within 30 days was performed in 30% of elderly patients and 48% of younger patients (P <.001). Elderly patients had a more unfavorable 30-day outcome compared with younger ones, as shown by the higher rates of death (6.4% vs 1.7%), acute myocardial infarction (7.1% vs 5%), and stroke (1.3% vs 0.5%). Multivariate analysis of the elderly group identified a conservative strategy (OR, 2.31; 95% CI, 1.20 to 4.48) and a diagnosis of non-Q wave MI (OR, 2.27; 95% CI, 1.32 to 3.93) as independent predictors of 30-day events. CONCLUSIONS: The elderly represent a very high-risk subgroup among patients with non-ST-elevation ACS, with a nearly 4-fold as high 30-day death rate as that of younger patients. These data call for a greater attention to such population, both in terms of an improved representation in clinical research and of the assessment of the outcome of different strategies in appropriately designed randomized trials. PMID- 15131539 TI - Diminishing proportional risk of sudden death with advancing age: implications for prevention of sudden death. AB - BACKGROUND: Advances in primary and secondary prevention of sudden death have led to a wide array of potentially beneficial therapies. Identification of patients most likely to benefit would be of use when considering costly interventions such as an implantable defibrillator. We sought to determine the effect of advancing age on the mode of death in the Amiodarone Trialists Metanalysis. METHODS AND RESULTS: Patients (n = 6252; age, 61.2+/-10.5 years; 83% men) were included in an analysis of predictors of sudden death (SD) and all-cause death (ACD), based on baseline variables at enrollment. Patients were divided into 5 age groups: < or =50 years, 51 to 60 years, 61 to 70 years, 71 to 80 years, and >80 years. During a mean of 16.8+/-10.3 months of follow-up, there were 1023 deaths, with an annual overall mortality rate of 11.7%. Both sudden death and nonsudden death rates increased with age, although the increase of nonsudden death with age was more dramatic. The overall proportion of death that was sudden (SD/ACD ratio) was 0.41, falling from 0.51 before age 50 years to 0.26 after age 80 years (P =.002 for trend). The SD/ACD ratio was not affected by sex, New York Heart Association Class, or left ventricular ejection fraction. CONCLUSIONS: Although the incidence of sudden death increases with age, the proportion of death that is sudden diminishes markedly. This finding may influence the yield of interventions targeted at prevention of sudden death. PMID- 15131540 TI - Obesity and the risk of death after acute myocardial infarction. AB - BACKGROUND: In the general population, obesity is associated with an increased risk of all-cause death. However, the importance of obesity in patients with established coronary heart disease is less well defined. METHODS: As part of the Determinants of Myocardial Infarction Onset Study, we performed a prospective cohort study of 1898 patients hospitalized with confirmed acute myocardial infarction between 1989 and 1994, with a median follow-up of 3.8 years. We assessed all-cause death through December 1995, using the National Death Index. We categorized patients according to WHO criteria for body mass index (BMI). We compared long-term death according to BMI (kg/m2) by using Cox proportional hazards regression. RESULTS: Of the 1898 eligible patients, 607 (32%) were normal weight (18.5 to 24.9 kg/m2), 832 (44%) were overweight (25.0 to 29.9 kg/m2), 331 (17%) were class I obese (30.0 to 34.9 kg/m2), and 128 (7%) were class II or more obese (> or =35.0 kg/m2). A total of 311 patients died during follow-up. After adjustment for potentially confounding risk factors and excluding patients with noncardiac comorbidity, the risk for death appeared to increase linearly, with increasing BMI across all categories (P for trend =.08). The relative risk of death in all obese patients (> or =30 kg/m2) was 1.46, compared with those with normal weight (95% CI, 0.98 to 2.17). CONCLUSIONS: We found that BMI appeared to have a positive, graded relation with post-myocardial infarction death. Whether weight reduction and secondary prevention strategies would reverse this effect in obese population remains to be seen. PMID- 15131541 TI - Association of the timing of ST-segment resolution with TIMI myocardial perfusion grade in acute myocardial infarction. AB - BACKGROUND: More complete ST-segment resolution (ST res) in acute myocardial infarction (MI) has been associated with better epicardial and myocardial reperfusion as assessed with the Thrombolysis in Myocardial Infarction (TIMI) flow grade (TFG) and the TIMI myocardial perfusion grade (TMPG), respectively. However, no data exist comparing the speed of ST resolution on continuous electrocardiogram (ECG) monitoring with the TMPG on coronary angiography. We hypothesized that delayed ST res is associated with impaired TMPGs. METHODS: Continuous 12-lead ECG recordings and 60-minute angiographic data were analyzed in 120 patients with acute MI who received tenectaplase monotherapy or combination therapy with low-dose tenectaplase and eptifibatide in the Integrilin and Tenecteplase in Acute Myocardial Infarction (INTEGRITI) trial. RESULTS: More rapid ST res on continuous ECG monitoring was associated with improved TMPGs on coronary angiography performed 60 minutes after study drug administration. For TMPG 3, the median time to ST resolution was 53 minutes. For TMPG 2, 1, and 0, the corresponding times were 64 minutes, 80 minutes, and 106 minutes, respectively (P =.01 for trend). Likewise, more rapid ST res was also associated with faster epicardial flow. For TFG 3, the median time to ST resolution was 46 minutes, compared with 109 minutes for TIMI flow grades 0 to 2 (P =.001). The corresponding times for a corrected TIMI frame count < or =40 versus >40 were 52 minutes and 112 minutes, respectively (P <.001). CONCLUSIONS: Although the static ECG has been associated with epicardial and myocardial blood flow in the past, this study extends these observations to demonstrate that more rapid ST res on continuous ECG monitoring is associated with improved myocardial perfusion after thrombolytic administration. PMID- 15131542 TI - ST-segment analyses and residual thrombi in the infarct-related artery: a report from the ASSENT PLUS ST-monitoring substudy. AB - BACKGROUND: Evolution of the ST segment during ST-elevation myocardial infarction (STEMI) has been shown to yield more information on prognosis than widely used invasive measurements. With continuous ST monitoring, even very occasional dynamic changes can be analyzed. We have recently suggested that ST variability during the reperfusion-phase is of prognostic importance. We wanted to further investigate this and relate it to angiographic findings. METHODS: A total of 177 patients with STEMI were examined in the ST-monitoring substudy of the ASessment of the Safety and Efficacy of a New Thrombolytic (ASSENT) PLUS trial, comparing dalteparin with heparin as adjunctive therapy to t-PA. Patients underwent 24 hours of ST monitoring. These recordings were blindly analyzed by 2 independent observers. A coronary angiogram was performed on days 4 to 7, also blindly evaluated by 2 persons. RESULTS: Occurrence of ST re-elevations during and after the reperfusion-phase was significantly associated with residual thrombi and TIMI flow in the infarct-related artery. Patients without any ST re-elevations showed a thrombus in only 5% of cases, as compared with 86% of patients with prolonged (lasting >30 minutes) ST re-elevations. In a multivariate comparison including baseline-data and treatment, most information on persistence of thrombi was contributed by the presence of any ST re-elevations (odds ratio, 5.8; 95% CI, 1.3 26). CONCLUSION: ST re-elevations during the first day of an acute myocardial infarction are associated with residual thrombi in the infarct-related artery even 4 to 7 days after the STEMI. PMID- 15131543 TI - Acute coronary syndromes complicated by symptomatic and asymptomatic heart failure: does current treatment comply with guidelines? AB - BACKGROUND: Patients with acute coronary syndromes (ACS) complicated by heart failure (HF) are at increased risk of death. Treatment with angiotensin converting enzyme inhibitors (ACEI), beta-blockers, and early invasive risk stratification are recommended for these patients. AIM: The purpose of the current study was to assess adherence to treatment guidelines of patients with ACS complicated by HF in Europe and the Mediterranean region. METHODS AND RESULTS: Of the 10,484 patients who participated in Euro-Heart ACS survey, 9587 had known HF status and were without cardiogenic shock; 7058 (74%) did not have symptomatic HF and 2529 (26%) presented with or developed symptomatic HF during hospitalization. HF patients were older and had more cardiovascular risk factors. ACEI were more commonly used in HF patients (75% vs 56%, P < .01), whereas beta blockers were less frequently used (75% vs 82%, P < .01). Coronary angiography and in hospital revascularization rates were lower among HF patients (42% vs 57% for coronary angiography, P < .01, and 32% vs 42% for revascularization, P < .01). Similar trends were noticed among patients with left ventricular dysfunction (symptomatic and asymptomatic).Adjusted in-hospital mortality risk was higher among patients with ACS complicated by symptomatic HF regardless of electrocardiographic type of ACS: (ST-elevation ACS, OR 2.5, 95% CI 1.6-3.9; non ST-elevation ACS, OR 8.9,95% CI 4.5-17.7; undetermined-ECG ACS, OR 9.3, 95% CI 2.5-34). CONCLUSIONS: Patients with ACS complicated by HF were at increased risk of dying. A relatively high percentage of HF patients were treated with ACEI and beta-blockers in accordance with current recommendations. Rates of coronary angiography and revascularization were significantly lower in ACS patients with HF versus those without HF, which potentially contributed to their worse mortality [corrected] PMID- 15131544 TI - Bleeding events with abciximab in acute coronary syndromes without early revascularization: An analysis of GUSTO IV-ACS. AB - BACKGROUND: The glycoprotein IIb/IIIa receptor antagonist abciximab reduces the risk of thrombotic complications with percutaneous coronary intervention, but also has been associated with higher bleeding rates. METHODS: In the Global Use of Strategies to Open Occluded Arteries in Acute Coronary Syndromes (GUSTO IV ACS) trial, abciximab (either a 24-hour or 48-hour infusion) was compared with placebo in 7800 patients with an acute coronary syndrome. During study drug administration, 2% of the patients underwent a revascularization procedure. RESULTS: In 1507 patients (19.3%), bleeding according to the Thrombolysis in Myocardial Infarction (TIMI) classification was observed while they were hospitalized or within 7 days. Ninety-eight patients (1.2%) had a major bleed, including 8 with intracranial hemorrhages. In 215 patients (2.8%), a minor bleed was reported, and in 1194 patients (15.3%), an insignificant bleed was reported. Bleeding was more frequent in patients receiving a 48-hour infusion of abciximab. Spontaneous bleeding was seen in 911 patients (11.7%). The other 596 patients had a bleeding event in conjunction with a procedure. The most significant predictors for bleeding with multivariable analysis were: use of low-molecular weight heparin, duration of abciximab infusion, region of hospitalization, performance of coronary artery bypass grafting or percutaneous coronary intervention (PCI), advanced age, and female sex. For major bleeding, the predictors were performance of coronary artery bypass grafting or PCI, long duration of abciximab administration, and advanced age. CONCLUSION: Treatment with abciximab in patients with non-ST-elevation acute coronary syndromes is safe because major bleeding and stroke are rare, and most events are clinically manageable or have few clinical consequences. Guidelines for use of abciximab in combination with other antithrombotic agents developed for PCI should also be respected in acute coronary syndromes. Specific dosing guidelines for combination with low-molecular weight heparin must be developed for patients who subsequently will undergo a PCI. PMID- 15131545 TI - Vascular basis for the treatment of myocardial ischemia study: trial design and baseline characteristics. AB - BACKGROUND: Increased low-density lipoprotein (LDL) and oxidized LDL cholesterol levels adversely affect endothelial function in patients with stable coronary artery disease (CAD). Statin drugs are efficacious in primary and secondary prevention of clinical CAD events, but they have not been extensively studied as a treatment for ischemia during routine daily activities or during exercise, indicators of high-risk in patients with stable CAD. The purpose of the Vascular Basis for the Treatment of Myocardial Ischemia study is to determine whether aggressive lowering of LDL cholesterol level with atorvastatin, with or without supplemental antioxidant vitamins C and E, can improve endothelial function and ischemia during ambulatory electrocardiogram (AECG) monitoring and exercise treadmill testing (ETT). METHODS: Patients are eligible when they have ischemia during an ETT and AECG monitoring and when their fasting total cholesterol level is < or =250 mg/dL. Eligible patients are randomized to receive 1 of 3 treatments: intensive atorvastatin to reduce LDL cholesterol level to < or =80 mg/dL, intensive atorvastatin to reduce LDL cholesterol level to < or =80 mg/dL plus antioxidant vitamins C and E, and control of diet and low-dose lovastatin, when needed, to reduce LDL cholesterol level < or = to 130 mg/dL. Patients undergo endothelial function testing, 48-hour AECG monitoring, and ETT at randomization and at 6 and 12 months. RESULTS: A total of 300 patients have been randomized: 101 to receive atorvastatin alone, 103 to receive atorvastatin plus antioxidant vitamins, and 96 to receive placebo. Baseline characteristics are similar across treatment groups. CONCLUSIONS: The Vascular Basis study will provide important insight on the effects of aggressive management of dyslipidemia with statin drugs and antioxidant vitamins in patients with stable but high-risk CAD. PMID- 15131546 TI - Long-term outcome of patients with vasovagal syncope. AB - BACKGROUND: After abnormal head-up tilt test (HUT), several trials have evaluated treatment strategies for vasovagal syncope (VVS). However, few unequivocal results have been obtained. The aim of the study was to prospectively analyze the natural history of patients with VVS who did not undergo specific treatment but received education for avoiding syncope after an abnormal HUT. METHODS: From 1996, 334 consecutive patients with VVS and an abnormal HUT result were followed. All of them received education for avoiding syncope as first-line therapy. RESULTS: During 30.4 +/- 21 months, there were no cardiac deaths. However, 101 patients (30.2%) had recurrences (1 recurrence, n = 64; > or =2, n = 37), which were not influenced by the type of response during HUT or by age. Time to first recurrence was correlated with the number of recurrences (r: -0.34, P =.0001). Mean recurrence-free time was 50.1 months (95% CI 46-54) and the cumulative probability of no recurrence was 69.8%. Receiver operator character curve analysis, demonstrated significant differences in the recurrence rate between patients with <5 or > or =5 previous episodes of syncope (25.1% vs 44%; P =.001). In addition, the mean recurrence-free time of patients with <5 episodes was significantly longer (54.1 months) than in patients with > or =5 episodes (39.6 months; P =.0005). Multivariate logistic regression identified the previous number of episodes as an independent risk marker of recurrences (odds ratio 2.34, 95% CI 1.4-3.8, P =.001). Age (at HUT performance) as a continuous variable was not a predictor of outcome. CONCLUSIONS: Survival of patients with VVS presenting after an abnormal HUT is excellent. Although the broad majority of those patients do not suffer recurrences after education, the number of previous syncopal episodes critically influences the recurrence rate. PMID- 15131547 TI - Yield of stress single-photon emission computed tomography in asymptomatic patients with diabetes. AB - BACKGROUND: Patients with diabetes without clinically apparent coronary artery disease are at increased risk of cardiac death. The value of screening stress testing in these patients remains controversial. The goal of this study was to examine the yield of stress single-photon emission computed tomography (SPECT) in asymptomatic diabetic patients. METHODS: The results of stress SPECT in patients without prior myocardial infarction or coronary revascularization were compared in asymptomatic diabetics (n = 1738) versus symptomatic diabetic patients (n = 2998), asymptomatic nondiabetic patients (n = 6215), and symptomatic nondiabetic patients (n = 16,214). RESULTS: Abnormal scans were present in 58.6% of asymptomatic diabetic patients, approximately equal to the percentage in symptomatic diabetic (59.5%) (P = not significant) and higher than in asymptomatic nondiabetic (46.2%) (P <.001) and symptomatic nondiabetic (44.4%) (P <.001) patients. The breakdown of high-risk scans followed a similar pattern in the 4 patient subsets: asymptomatic diabetic, 19.7% versus symptomatic diabetic, 22.2% (P =.051); asymptomatic nondiabetic, 11.1% (P <.001); and symptomatic nondiabetic, 12.5% (P <.001). Patients with diabetes had more electrocardiographic and scan evidence for silent myocardial infarction versus those without diabetes. CONCLUSIONS: Asymptomatic diabetic patients have a high prevalence of both abnormal and high-risk SPECT scans. The finding that approximately 1 in 5 of these individuals has a high-risk scan suggests a potentially more widespread application of screening stress SPECT in asymptomatic diabetic patients to identify those with severe coronary artery disease. PMID- 15131548 TI - Prevalence of resistance against activated protein C resulting from factor V Leiden is significantly increased in myocardial infarction: investigation of 507 patients with myocardial infarction. AB - BACKGROUND: A point mutation in the gene encoding coagulation factor V is a cause of resistance against activated protein C. The presence of factor V Leiden is linked to 50% of congenital defects causing venous thrombosis. Its relationship to arterial thrombosis, particularly to myocardial infarction, has not been defined. Therefore, we performed a study on the role of factor V Leiden in patients with myocardial infarction. The study was carried out in Bavarians of German origin, a relatively homogeneous population. METHODS AND RESULTS: The study group consisted of 507 patients with documented myocardial infarction (77.5% (393/507) men, 22.5% (114/507) women), with a mean age of 56.1 (range 18 86) years. Strict criteria for patient selection and highly sensitive and specific functional tests for factor V Leiden were used. In addition, all patients with pathological test results were genotyped. The prevalence of factor V Leiden in patients with myocardial infarction was 8.7% (44/507), a significant increase in the prevalence of this mutation compared with the control group (3.7%, P =.0025). The odds ratio was 2.46 (95% CI 1.35-4.50). CONCLUSIONS: A significantly increased prevalence of factor V Leiden in patients with documented myocardial infarction was seen. Patients with this mutation appear to have a predisposition for myocardial infarction. PMID- 15131549 TI - Replication of the association between the thrombospondin-4 A387P polymorphism and myocardial infarction. AB - BACKGROUND: The purpose of our study was to replicate an association between the A387P polymorphism in the thrombospondin-4 (THBS4) gene and myocardial infarction (MI), as previously reported by our group while taking confounding into account, and to assess whether ascertainment by age of onset would affect this association. METHODS: We performed a case-control study of 474 white patients with MI (not selected on the basis of age of onset nor family history) and 472 white control subjects. We then applied our findings to our original population of 184 white patients with premature, familial MI and 406 white control subjects. RESULTS: In the replication population, no significant association was found between THBS4 genotype and MI (P =.41) with univariate analysis. However, after adjusting for age, sex, first-degree family history, and waist-to-hip ratio, an association was apparent in the replication population (P =.032), and the original association became much stronger (P =.00008). Both studies showed a 2.5- to 3-fold increased odds of MI in individuals with the P allele. Furthermore, several variables appeared to modify the effect of THBS4 on MI, including waist to-hip ratio, diabetes mellitus, and hypertension. When we stratified our cases by age of onset (< or =45 years in men, < or =50 years in women), there were no significant differences in genotype frequencies when comparing premature cases with late-onset cases or premature cases with control subjects in either unadjusted or adjusted analyses (all P values >.25). CONCLUSIONS: Our findings suggest that the A387P variant of the THBS4 gene may be an important determinant in the development of MI at any age. Careful assessment of clinical covariates helped to unmask a significant association and therefore may be an important reason for why studies do not replicate. PMID- 15131550 TI - Recovery kinetics of oxygen uptake is prolonged in adults with an atrial septal defect and improves after transcatheter closure. AB - BACKGROUND: In adults with an atrial septal defect (ASD) transcatheter closure leads to an improvement of peak oxygen uptake (VO2), but the kinetics of recovery of VO2 after maximal exercise in this patient population and the impact of transcatheter ASD closure have never been investigated. METHODS: Twenty consecutive patients underwent a maximal cardiopulmonary exercise test both the day before and 6 months after transcatheter ASD closure. For comparison, an age- and sex-matched group consisting of 53 healthy adults was built. The constant decay of VO2, CO2 production (VCO2), minute ventilation (VE), and heart rate (HR), expressed as the first-degree slope of a single linear relation, were calculated for the first minute of recovery. RESULTS: Patients with an ASD had a prolonged VO2 slope (P =.0012), VCO2 slope (P =.0003), and VE slope (0.013) when compared with control subjects. Six months after transcatheter ASD closure, significant improvements of VO2 slope (P =.0043) and of VCO2 slope (P =.0022) were recorded, so that no difference was found when compared with those of the control group (P =.1 and P =.06, respectively). The VE slope and HR slope did not change after closure. A significant association between VO2 slope and peak VO2 in the group of patients with ASD was shown by the Spearman correlation, both before (r = 0.67, P =.0012) and after ASD closure (r = 0.71, P =.0004). CONCLUSIONS: A limited cardiopulmonary reserve in adults with no symptom who have an ASD appears to affect not only maximal exercise responses but also the recovery phase. Transcatheter ASD closure induces a significant improvement of the ability of recovering from maximal exercise and eliminates the difference with a healthy population. PMID- 15131551 TI - Lipid profile of patients with aortic stenosis might be predictive of rate of progression. AB - BACKGROUND: Aortic stenosis is one of the most commonly encountered valvular pathology requiring surgery in developed countries. There are similarities between risk factors for coronary atherosclerosis and the development of aortic stenosis. We designed a retrospective study, evaluated the lipid profile and previous echocardiographic recordings of patients with aortic stenosis, and searched the association of rate of progression and lipid profile. METHODS AND RESULTS: The annual rates of progression in the peak and mean aortic gradients were 8.5 +/- 3.2 and 6.7 +/- 2.2 mm Hg/year, respectively. We classified the annual rate of progression of peak aortic gradient into 2 groups, group 1 with <10 mm Hg ("slow progressors") and group 2 with > or =10 mm Hg annual rate of progression ("fast progressors"). The annual rate of progression in group 1 was significantly higher than that in group 2, both in peak and mean aortic gradients (12 +/- 2 mm Hg and 6.4 +/- 1.6 mm Hg; 9 +/- 1.3 mm Hg and 5.2 +/- 1.1 mmHg; P <.001 for both). There was a highly significant difference between group 1 and group 2 for total cholesterol/high-density lipoprotein (HDL) cholesterol level ratio (7.1 +/- 1.4 vs 5.2 +/- 1.3, P <.001). There was a significant correlation between annual rate of progression in peak gradient and total cholesterol/HDL cholesterol level ratio (r = 0.399, P =.009). Smoking (P =.024, Beta = 0.26), presence of coronary heart disease (P =.011, Beta = 0.31), and total cholesterol/HDL cholesterol level ratio (P =.004, Beta = 1.98) were independently predictive of fast progression of the peak aortic gradient in the regression analysis. CONCLUSION: In a small group of patients from Turkey with aortic stenosis, there seems to be an association between the rate of progression and total cholesterol/HDL cholesterol level ratio, with fast progression occurring in the group with higher ratios. PMID- 15131552 TI - Endomyocardial biopsy plays a role in diagnosing patients with unexplained cardiomyopathy. AB - BACKGROUND: The etiology of cardiomyopathy is usually inferred from clinical information and preliminary laboratory studies. Patients with unexplained cardiomyopathy may be referred for endomyocardial biopsy (EMBx). It is unknown whether pathological information obtained from EMBx is beneficial or alters the diagnosis established clinically. This study was undertaken to evaluate the utility of EMBx in confirming or excluding a clinically suspected diagnosis. METHODS: We evaluated 845 patients with initially unexplained cardiomyopathy who underwent EMBx between 1982 and 1997 at The Johns Hopkins Hospital. For each patient, an initial clinical diagnosis, an EMBx diagnosis, and a final diagnosis prior to discharge based on all available data were established. RESULTS: The final diagnosis differed from the initial clinical diagnosis in 264 (31%) of these patients; EMBx made the diagnosis in 196 (75%) of these cases. Initial diagnoses most frequently altered were myocarditis (34%) and idiopathic cardiomyopathy (25%). Initial diagnoses least likely to be altered were those in which biopsy was used to confirm or grade a previously documented illness, such as hemochromatosis (11%), amyloidosis (18%), or cardiomyopathy secondary to doxorubicin toxicity (0%). EMBx was more sensitive than clinical diagnosis in detecting myocarditis and amyloidosis, and proved to be very specific in detecting ischemic cardiomyopathy, myocarditis, amyloidosis, and hemochromatosis. CONCLUSIONS: In patients with unexplained cardiomyopathy after a standard evaluation, the clinical assessment of the etiology is inaccurate in 31% of patients. EMBx establishes the final diagnosis in 75% of these patients with a high degree of specificity. PMID- 15131553 TI - Are hematinic deficiencies the cause of anemia in chronic heart failure? AB - BACKGROUND: Anemia in chronic heart failure (CHF) is common, varying in prevalence between 14.4% and 55%, and is more frequent in patients with more severe heart failure. Patients with CHF who have anemia have a poorer quality of life, higher hospital admission rates, and reduced exercise tolerance. We explored the relation between hematinic levels and hemoglobin (Hb) levels and exercise tolerance in a group of patients with CHF. METHODS: We analyzed data from 173 patients with left ventricular systolic dysfunction (LVSD), 123 patients with symptoms of heart failure, but preserved left ventricular (LV) systolic function ("diastolic dysfunction"), and 58 control subjects of similar age. Each underwent echocardiography, a 6-minute walk test, and blood tests for renal function and Hb and hematinic levels (vitamin B12, iron, and folate). We classified patients as having no anemia (Hb level >12.5 g/dL), mild anemia (Hb level from 11.5-12.5 g/dL), or moderate anemia (Hb level <11.5 g/dL). RESULTS: Of patients with LVSD, 16% had moderate anemia and 19% had mild anemia. Of patients with preserved LV function, 16% had moderate anemia and 17% had mild anemia. Four control subjects had a Hb level <12.5 g/dL. Of all patients, 6% were vitamin B12 deficient, 13% were iron deficient, and 8% were folate deficient. There was no difference between patients with LVSD and the diastolic dysfunction group. In patients with LVSDS, the average Hb level was lower in New York Heart Association class III than classes II and I. The distance walked in 6 minutes correlated with Hb level in both groups of patients with CHF (r = 0.29; P <.0001). Patients with anemia achieved a lower pVO2 (15.0 [2.3] vs 19.5 [4.4], P <.05). Peak oxygen consumption correlated with Hb level (r = 0.21, P <.05) in the patients, but not in the control subjects. In patients with anemia, the mean creatinine level was higher than in patients with a Hb level >12.5 g/dL, but there was no clear relationship with simple regression. Hematocrit level and mean corpuscular volume were not different in the patients with diastolic dysfunction, patients with LV dysfunction, or the control subjects. Hematocrit levels were not influenced by diuretic dose. Patients with anemia were not more likely to be hematinic deficient than patients without anemia. CONCLUSIONS: Patients with symptoms and signs of CHF have a high prevalence of anemia (34%) whether they have LV dysfunction or diastolic dysfunction, but few patients have hematinic deficiency. Hemoglobin levels correlate with subjective and objective measures of severity and renal function. PMID- 15131554 TI - Independent prognostic value of elevated high-sensitivity C-reactive protein in chronic heart failure. AB - BACKGROUND: The serum concentration of C-reactive protein (CRP) is mildly elevated in patients with chronic congestive heart failure (CHF), but this level falls well within the range found in healthy subjects. Standard clinical assays for CRP lack sensitivity within the low reference range and thus cannot be used effectively for routine clinical risk prediction. Because assays for high sensitivity CRP (hsCRP) are now available, we can measure hsCRP to determine its predictive value for the prognosis of patients with CHF. METHODS: Serum levels of hsCRP in 108 patients with CHF and left ventricular ejection fraction (LVEF) <50% were examined. Major adverse cardiac events (death, heart transplantation, or hospitalization with worsening heart failure) during a median follow-up period of 403 days were determined. RESULTS: The concentrations of hsCRP in this study population were significantly increased with the severity of CHF. In a multivariate analysis, LVEF and serum levels of hsCRP were independent significant predictors for adverse outcomes in these patients (hazard ratio, 3.714, P =.024, and hazard ratio, 2.584, P =.047, respectively). However, hsCRP was minimally correlated with LVEF (r = -0.167, P =.084). Further analysis indicated that hsCRP might identify a different high-risk group and could improve risk stratification beyond that of LVEF. CONCLUSIONS: These findings suggest that an elevated level of hsCRP is an independent predictor of prognosis in CHF and can provide additional prognostic information for the risk stratification and treatment in patients with chronic CHF. PMID- 15131555 TI - A randomized trial comparing monophasic and biphasic waveform shocks for external cardioversion of atrial fibrillation. AB - BACKGROUND: We compared efficacy of and pain felt after biphasic truncated exponential (BTE) and monophasic damped sine (MDS) shocks in patients undergoing external cardioversion of atrial fibrillation (AF). METHODS: Patients with AF were randomized to BTE or MDS waveform cardioversion. Successive shocks were delivered at 70, 100, 200, and 360 J until successful cardioversion, with one 360 J attempt of the alternate waveform when all 4 shocks failed. Success was determined by blinded over-read of electrocardiograms. Peak current was calculated from energy and impedance. Patients rated their pain at 1 and 24 hours after cardioversion. RESULTS: Fourteen of 37 (38%) patients treated with MDS and 34 of 35 (97%) treated with BTE shocks were cardioverted at < or =200 J (P <.0001). Success rates of MDS versus BTE shocks were 5.4% versus 60% for 70 J, 19% versus 80% for < or =100 J, and 86% versus 97% for < or =360 J. BTE shocks cardioverted with less peak current (14.0 +/- 4.3 vs 39.5 +/- 11.2 A, P <.0001), less energy (97 +/- 47 vs 278 +/- 120 J, P <.0001), and less cumulative energy (146 +/- 116 vs 546 +/- 265 J, P <.0001). Patients felt less pain after BTE than MDS shocks at 1 hour (P <.0001) and 24 hours (P <.0001) after cardioversion. CONCLUSION: This BTE waveform is superior to the MDS waveform for cardioversion of AF, requiring much less energy and current, and causing less postprocedural pain. PMID- 15131556 TI - Eight weeks of moderate-intensity exercise training increases heart rate variability in sedentary postmenopausal women. AB - BACKGROUND: Regular exercise is associated with increased heart rate variability (HRV). However, results from studies examining the effect of exercise training on HRV in postmenopausal women are inconclusive. In addition, the effect of hormone replacement therapy (HRT) on HRV remains a subject of speculation. METHODS: We examined 88 sedentary postmenopausal women in a randomized controlled trial who were assigned to exercise (n = 49) or control (n = 39) groups. The exercising women performed 8 weeks of aerobic exercise training at a heart rate equivalent to 50% of VO2max, consisting on average of 44 minutes per session, 3 to 4 times per week. Resting HRV was measured in each participant at baseline and after 8 weeks of intervention. Ten minutes of resting R-R intervals were analyzed by time (standard deviation of mean R-R intervals, root of mean square successive differences) and frequency domain methods: low-frequency (LF) was defined as 0.04 to 0.15 Hz, high-frequency (HF) as 0.15 to 0.40 Hz, and total spectral power as 0.00 to 0.40 Hz. The LF and HF components in normalized units were also calculated. RESULTS: At baseline, there were no significant differences in HRV between control and exercise groups. Additionally, there were no differences in any HRV variables when women were grouped by HRT use (no HRT, estrogen-only HRT, and progestin-containing HRT). After 8 weeks, women randomly assigned to the exercise group increased all absolute time and frequency domain indexes (all P <.001) and reduced resting heart rate (P =.002) compared with women in the control group. The LF and HF components expressed as normalized units remained unchanged after exercise intervention. Additionally, HRT use did not modify the exercise-induced changes in HRV. CONCLUSIONS: We conclude that moderate aerobic exercise increases HRV in sedentary postmenopausal women. This benefit is not influenced by the use of HRT. PMID- 15131557 TI - Primary stenting of occluded native coronary arteries: final results of the Primary Stenting of Occluded Native Coronary Arteries (PRISON) study. AB - BACKGROUND: Primary intracoronary stent placement after successfully crossing chronic total coronary occlusions may decrease the high restenosis rate at long term follow-up compared with conventional balloon angioplasty. METHODS: In a prospective, randomized trial, balloon angioplasty was compared with stent implantation for the treatment of chronic total occlusions. Patients were followed for 12 months with angiographic follow-up at 6 months. Quantitative coronary analysis was performed by an independent core lab. RESULTS: A total of 200 patients were enrolled. Baseline characteristics were evenly distributed. After the procedure the mean minimal luminal diameter in the conventional group was 2.34 +/- 0.46 mm versus 2.90 +/- 0.41 mm in the stented group (P <.0001). The 6-month angiographic follow-up showed a mean minimal luminal diameter of 1.57 +/- 0.74 mm in the conventional group versus 1.93 +/- 0.85 mm in the stented group (P =.009) and a mean diameter stenosis of 44.7% +/- 25.0% versus 35.5% +/- 26.5% (P =.036). Binary angiographic restenosis (>50% diameter stenosis) was seen in 33% in the conventional group versus 22% in the stented group (P =.137). The reocclusion rates were 7.3% and 8.2%, respectively (P = 1.00). At 12 month follow up, the rate of target lesion revascularization was significantly higher in the conventional group (29% versus 13%, P <.0001). CONCLUSION: These data demonstrate that stenting of chronic total occlusions is superior to balloon angioplasty alone with a statistically significant reduction in the need for target lesion revascularization and a lower, but not significant, restenosis rate. PMID- 15131558 TI - Effect of rosiglitazone on restenosis after coronary stenting in patients with type 2 diabetes. AB - BACKGROUND: Thiazolidinediones have been shown to have an antiproliferative vascular effect in experimental models. We sought to study the effect of rosiglitazone on in-stent restenosis in patients with established type 2 diabetes. METHODS: Patients with treated type 2 diabetes (mean duration 5.5 +/- 7.5 years) referred for coronary stenting were randomized in a double-blind fashion to receive oral rosiglitazone or placebo for 6 months. Quantitative coronary angiography and intravascular ultrasound data were obtained at baseline and follow-up. Plasma plasminogen activator inhibitor-1 levels were prospectively measured. RESULTS: Sixteen patients were enrolled. There were no significant differences in follow-up in-stent luminal diameter stenosis measured by quantitative coronary angiography or in-stent luminal area stenosis and neointimal volume index obtained by intravascular ultrasound, nor were there any differences in plasma plasminogen activator inhibitor-1 levels after long-term use despite improvement in diabetes control and insulin sensitivity. CONCLUSIONS: Rosiglitazone, given at the time of stent implantation in treated diabetics, did not reduce in-stent restenosis in this small series. The vascular biological effects of this agent await further clarification in humans and evaluation in larger clinical trials. PMID- 15131559 TI - Effect of a cardiac rehabilitation program on left ventricular diastolic function and its relationship to exercise capacity in patients with coronary heart disease: experience from a randomized, controlled study. AB - BACKGROUND: A cardiac rehabilitation and prevention program (CRPP) is a recognized nonpharmacological modality in the management of coronary heart disease (CHD). However, the effect of a CRPP on systolic function of the heart is controversial, and no data exists on diastolic function in CHD. A randomized, controlled study was conducted to address these issues. METHODS: Patients (n = 269) with recent acute myocardial infarction (n = 193) or after percutaneous coronary intervention (PCI) (n = 76) were randomized to either CRPP (2-hour twice weekly exercise program for 8 weeks) or conventional therapy (control group). Serial treadmill exercise testing and at-rest echocardiography were performed during phases 1 (baseline), 2 (post-exercise training), and 3 (8-month follow up). RESULTS: The prevalence of left ventricular (LV) abnormal relaxation pattern (ARP) of diastolic dysfunction was increased in the control group only in phase 3 (65% vs 88%, chi2 = 7.6, P <.01). Significant improvement of individual LV diastolic parameters towards less severe delayed relaxation was also observed in the CRPP group, especially in those with recent acute myocardial infarction or ARP. The gain in exercise capacity was faster and more substantial in the CRPP than the control group (P <.001 for phase 2, P <.05 for phase 3), and was significantly correlated with LV diastolic indices in those with ARP. Exercise training had neutral effects on LV systolic function and rate-pressure product. CONCLUSIONS: In patients with CHD, CRPP prevented the progression of resting LV diastolic dysfunction, without affecting systolic function. In those with ARP, the improvement of diastolic function predicted the gain in exercise capacity. PMID- 15131561 TI - Update on food allergy. AB - Tremendous progress has been made in our understanding of food-based allergic disorders over the past 5 years. Recent epidemiologic studies suggest that nearly 4% of Americans are afflicted with food allergies, a prevalence much higher than appreciated in the past. In addition, the prevalence of peanut allergy was found to have doubled in American children less than 5 years of age in the past 5 years. Many food allergens have been characterized at the molecular level, which has contributed to our increased understanding of the immunopathogenesis of many allergic disorders and might soon lead to novel diagnostic and immunotherapeutic approaches. The management of food allergies continues to consist of educating patients on how to avoid relevant allergens, to recognize early symptoms of an allergic reaction in case of an accidental ingestion, and to initiate the appropriate emergency therapy. However, the recent successful clinical trial of anti-IgE therapy in patients with peanut allergy and the number of immunomodulatory therapies in the pipeline provide real hope that we will soon be able to treat patients with food allergy. PMID- 15131562 TI - A classification of plant food allergens. AB - Plant food allergens can be classified into families and superfamilies on the basis of their structural and functional properties. The most widespread groups of plant proteins that contain allergens are the cupin and prolamin superfamilies and the protein families of the plant defense system. The cupin superfamily includes allergenic seed storage proteins of the vicilin and legumin type present in soybeans, peanuts, and tree nuts. The prolamin superfamily includes several important types of allergens of legumes, tree nuts, cereals, fruits, and vegetables, such as the 2S albumin seed storage proteins, the nonspecific lipid transfer proteins, and the cereal alpha-amylase and protease inhibitors. Plant food allergens are also found among the various groups of defense proteins that enable plants to resist biotic and abiotic stress, such as the pathogenesis related proteins, certain proteases, and protease inhibitors. This review focuses on a classification system of plant food allergens that is emerging from the synopsis of allergology and protein evolution. PMID- 15131563 TI - Revised nomenclature for allergy for global use: Report of the Nomenclature Review Committee of the World Allergy Organization, October 2003. AB - The nomenclature proposed in the October 2003 report of the Nomenclature Review Committee of the World Allergy Organization is an update of the European Academy of Allergology and Clinical Immunology Revised Nomenclature for Allergy Position Statement published in 2001. The nomenclature can be used independently of target organ or patient age group and is based on the mechanisms that initiate and mediate allergic reactions. It is assumed that as knowledge about basic causes and mechanisms improves, the nomenclature will need further review. PMID- 15131564 TI - First-aid treatment of anaphylaxis to food: focus on epinephrine. AB - Avoiding food triggers for anaphylactic reactions (severe acute systemic allergic reactions) is easier said than done. Most episodes of anaphylaxis to food occur unexpectedly in the community in the absence of a health care professional. All individuals at risk should therefore have an emergency action plan in place. The cornerstone of first-aid treatment of anaphylaxis is epinephrine injected intramuscularly in the vastus lateralis muscle (lateral aspect of the thigh). In this review, we focus on epinephrine. We examine a therapeutic dilemma: the issue of epinephrine dose selection in an individual for whom no optimal fixed-dose auto-injector formulation exists, and a therapeutic controversy: the issue of epinephrine injection versus an oral H1-antihistamine in anaphylaxis episodes that appear to be mild. The pharmaceutical industry could address the first of these issues by providing a wider range of epinephrine fixed doses in easy-to-use auto-injectors, or by providing adjustable epinephrine doses in auto-injectors. The second issue could be addressed in part by development of alternative routes of epinephrine administration for the first-aid, out-of-hospital treatment of anaphylaxis. PMID- 15131565 TI - Influence of early life exposures on incidence and remission of asthma throughout life. AB - BACKGROUND: Knowledge of the effects of early environmental and congenital factors on the natural history of asthma may provide important clues to the pathogenesis of asthma. OBJECTIVE: We assessed associations between potential, early determinants and the incidence and remission of asthma throughout life, and tested whether the strength and direction of these associations varied in childhood, adolescence, and adulthood. METHODS: The data pertaining to the individual asthma history of 18,156 subjects, age 0 to 44 years, who attended the clinical stage of the European Community Respiratory Health Survey were analyzed retrospectively by life-event methods. Onset of asthma was defined as age at the first attack, and asthmatic patients were considered to be in remission if they had not been under treatment or had an attack of asthma in the past 24 months. Onset and remission were evaluated in 3 time windows: <10, 10 to 20, and > or =20 years of age. The associations of asthma with early determinants were estimated by hazard ratios (HRs). RESULTS: A family history of asthma or allergy was associated with a higher risk of developing asthma (HR, 1.89; 95% CI, 1.67-2.13) and a lower chance of remission (HR, 0.79; 95% CI, 0.64-0.99) throughout life. No matter what one's genetic predisposition was, early, acute respiratory infections were associated with an increased lifelong risk of asthma onset (pooled HR, 3.19; 95% CI, 2.75-3.69), whereas early contact with older children, which is a marker of prolonged, intermittent exposure to infectious agents, conferred permanent protection against asthma (HR, 0.84; 95% CI, 0.74-0.96) and increased the chance of remission in childhood asthma (HR, 1.50; 95% CI, 1.10-2.04). Pet ownership had a protective effect only in childhood (HR, 0.78; 95% CI, 0.74-0.96), whereas maternal smoking did not show a significant association with asthma. Female sex was negatively associated with the onset of asthma in childhood (HR, 0.62; 95% CI, 0.52-0.75) and positively in adulthood (HR, 2.01; 95% CI, 1.61-2.51). The pattern of associations was similar in sensitized (positive assay to specific IgE) and nonsensitized asthmatic patients. CONCLUSION: Genetic predisposition and exposure to infectious agents are major early determinants that influence a subsequent history of asthma. The length and type of exposure to infectious agents seem able either to promote or to suppress an anti-inflammatory process, unrelated to IgE, which can partially interfere with an acquired predisposition for asthma. PMID- 15131566 TI - Treatment of asthma with nebulized lidocaine: a randomized, placebo-controlled study. AB - BACKGROUND: In 2 prior uncontrolled studies, nebulized lidocaine reduced oral glucocorticoid use in patients with severe glucocorticoid-dependent asthma. OBJECTIVE: We tested the safety and efficacy of nebulized lidocaine in a randomized, placebo-controlled study in patients with mild-to-moderate asthma. METHODS: We recruited 50 subjects (25 receiving lidocaine and 25 receiving placebo); all had a prebronchodilator FEV(1) of 64% to 125% of predicted normal value and were treated with daily inhaled glucocorticoids (but not systemic glucocorticoids) and bronchodilators for at least 2 months. Before treatment, subjects monitored their symptoms and peak flow values and maintained their medications for 2 weeks. At initiation, subjects inhaled either nebulized placebo (saline) or lidocaine (4%, 100 mg) 4 times daily. All subjects were instructed to reduce their inhaled glucocorticoid dosage by one half each week for 3 weeks and to discontinue glucocorticoid treatment at week 4. The subjects continued the nebulized lidocaine or placebo for a total of 8 weeks, monitored their symptoms, and used bronchodilators to control symptoms. RESULTS: Indicators of asthma severity showed benefit for the lidocaine-treated group: changes in FEV(1) (P < or =.001), nighttime awakenings (P < or =.02), symptoms (P < or =.010), bronchodilator use (P < or =.010), and blood eosinophil counts (P < or =.020). Subjects in both groups reduced use of inhaled glucocorticoids comparably. Subjects receiving nebulized placebo showed increases in their symptom scores, bronchodilator use (P < or =.05 for both), and blood eosinophil counts (P < or =.01) and decreases in FEV(1) (P < or =.001). CONCLUSION: Nebulized lidocaine provided effective and safe therapy in subjects with mild-to-moderate asthma. PMID- 15131567 TI - Microbial exposure of rural school children, as assessed by levels of N-acetyl muramic acid in mattress dust, and its association with respiratory health. AB - BACKGROUND: Endotoxin exposure has been shown to be associated with a decreased prevalence of atopic sensitization and symptoms. Yet endotoxin represents only a part of the indoor microbial exposure. Muramic acid, a constituent of peptidoglycan, is present in gram-negative and gram-positive bacteria in the environment and may therefore serve as an additional marker of microbial exposure. OBJECTIVE: To study the factors determining the level of indoor exposure to muramic acid/peptidoglycan, as well as its potential association with respiratory health. METHODS: In 553 farm and nonfarm school children from Austria, Switzerland, and Germany, mattress dust muramic acid concentrations were determined, and health was assessed by using IgE measurements and questionnaire information. RESULTS: The muramic acid concentration was found to be significantly higher in dust from farm children's mattresses than in dust from nonfarm children's mattresses (157 vs 131 ng/mg). Children with higher mattress dust muramic acid concentrations had a significantly lower prevalence of wheezing (odds ratio of highest vs lowest tertile of muramic acid concentration, 0.3; 95% CI, 0.1-0.9), regardless of farming status and endotoxin exposure. The association for asthma was similar, and no association was found with atopic sensitization. CONCLUSION: Next to endotoxin, muramic acid provides us with an independent marker of microbial exposure. Unlike endotoxin, muramic acid was inversely associated with wheezing rather than with atopic sensitization. PMID- 15131568 TI - Advances in mechanisms of allergy. AB - This review summarizes selected Mechanisms of Allergy articles appearing between 2002 and 2003 in the Journal of Allergy and Clinical Immunology. Articles chosen include those dealing with human airways disease pathophysiology, pharmacology, cell biology, cell recruitment, and genetics, as well as information from allergen challenge models in both human and nonhuman systems. When appropriate, articles from other journals have been included to supplement the topics being presented. PMID- 15131569 TI - Increased sensitivity of asthmatic airway smooth muscle cells to prostaglandin E2 might be mediated by increased numbers of E-prostanoid receptors. AB - BACKGROUND: An increase in airway smooth muscle (ASM) cell proliferation leads to an increase in the bulk of the ASM, one of the characteristic features of asthma. We have previously shown that ASM cells from asthmatic individuals proliferate more than those from nonasthmatic subjects. This increased growth might be due to compromised inhibitory mechanisms within the ASM of asthmatic subjects. OBJECTIVE: The purpose of this study was to determine whether the proliferative control exerted by prostaglandin E(2) (PGE(2)) was altered in the asthmatic ASM cells. METHODS: We used tritated thymidine uptake to measure cell proliferation and cell-surface ELISAs to detect the presence of cell-surface receptors on ASM cells isolated from asthmatic and nonasthmatic individuals. RESULTS: The asthmatic ASM cells were significantly more sensitive to proliferation inhibition by PGE(2) than the nonasthmatic cells (P<.02). The PGE(2) (E-prostanoid [EP]) receptors EP2 and EP3 were detected on asthmatic and nonasthmatic smooth muscle cells in culture. There were significantly more receptors on the asthmatic cells. The asthmatic cells also had increased sensitivity to proliferation inhibition by EP2-specific agonists but not by EP3-specific agonists. CONCLUSION: The increased growth observed in asthmatic ASM cells is not the result of impaired responsiveness to PGE(2). In contrast, these cells have increased sensitivity. This increased sensitivity might be mediated by the increased numbers of EP2 receptors on the surface. PMID- 15131570 TI - Peroxisome proliferator-activated receptor gamma is expressed in airways and inhibits features of airway remodeling in a mouse asthma model. AB - BACKGROUND: Allergic asthma is associated with persistent functional and structural changes in the airways and involves many different cell types. Peroxisome proliferator-activated receptor gamma (PPAR-gamma), a member of the nuclear hormone receptor superfamily, is predominantly expressed in adipose tissue and plays a major role in regulating adipocyte differentiation and glucose metabolism. Recently, PPAR-gamma has been shown to play an important role in the control of inflammatory responses, including within the lung, acting on both immune and nonimmune cells. OBJECTIVE: Our aim was to assess the anti inflammatory potential of a PPAR-gamma agonist locally delivered by means of nebulization. METHODS: We used a mouse model of asthma induced by sensitization and airway challenge with ovalbumin. Ciglitazone, a PPAR-gamma agonist, was administered by means of nebulization alone at the time of antigen challenge or by means of gavage and nebulization. Treatments with both ciglitazone and GW9662, a specific antagonist, were also performed to verify that ciglitazone's effects were mediated through PPAR-gamma activation. RESULTS: Our results show that PPAR gamma is mainly expressed in airway epithelium on antigen sensitization. Treatment with ciglitazone reduced PPAR-gamma levels in the lung, whereas combined treatment with GW9662 abrogated this inhibition. Importantly, nebulization with ciglitazone decreased airway hyperresponsiveness, basement membrane thickness, mucus production, collagen deposition, and TGF-beta synthesis. A significant correlation was also found between airway hyperresponsiveness, basement membrane thickness, and TGF-beta levels. CONCLUSION: These results demonstrate that inhaled agonistic ligands of PPAR gamma might have new therapeutic potential for airway asthmatic inflammation. PMID- 15131571 TI - Characterization of two polymorphisms in the leukotriene C4 synthase gene in an Australian population of subjects with mild, moderate, and severe asthma. AB - BACKGROUND: The cysteinyl-leukotrienes (cys-LTs) are proinflammatory mediators that are important in the pathophysiology of asthma. LTC(4) synthase is a key enzyme in the cys-LT biosynthetic pathway, and studies in small populations have suggested that a promoter polymorphism (A(-444)C) in the gene might be associated with asthma severity and aspirin intolerance. OBJECTIVE: We sought to screen the LTC(4) synthase gene for polymorphisms and to determine whether there is an association between these polymorphisms and asthma severity or aspirin sensitivity in a large, well-phenotyped population and to determine whether this polymorphism is functionally relevant. METHODS: The coding regions of the LTC(4) synthase gene were screened for polymorphisms and the A(-444)C polymorphism was analyzed in a large Australian white adult population of mild (n=282), moderate (n=236), and severe asthmatic subjects (n=86) and nonasthmatic subjects (n=458), as well as in aspirin-intolerant asthmatic subjects (n=67). The functional activity of the promoter polymorphism was investigated by transient transfection of HL-60 cells with a promoter construct. RESULTS: A new polymorphism was identified in intron 1 of the gene (IVS1-10c>a) but was not associated with asthma. Association studies showed that the A(-444)C polymorphism was weakly associated with asthma per se, but there was no association between the C(-444) allele and chronic asthma severity or aspirin intolerance. A meta-analysis of all the genetic studies conducted to date found significant between-study heterogeneity in C(-444) allele frequencies within different clinical subgroups. In vitro functional studies showed no significant differences in transcription efficiency between constructs containing the A(-444) allele or the C(-444) allele. CONCLUSIONS: Our data confirm that, independent of transcriptional activity, the C(-444) allele in the LTC(4) synthase gene is weakly associated with the asthma phenotype, but it is not related to disease severity or aspirin intolerance. PMID- 15131572 TI - Multilocus haplotype analyses reveal association between 5 novel IL-15 polymorphisms and asthma. AB - BACKGROUND: IL-15 is a T(H)1-related cytokine that is involved in the inflammatory response in various infectious and autoimmune diseases. IL-15 has recently been shown to be upregulated in T-cell-mediated inflammatory disorders. The observations suggest a potential role for this cytokine in a variety of pathologic conditions, including T(H)1-mediated and T(H)2-mediated inflammatory diseases. OBJECTIVE: In this study, we searched for single nucleotide polymorphisms in the whole IL-15 gene and investigated their association with inflammatory and/or atopic phenotypes. METHODS: The screening for single nucleotide polymorphisms was performed by single-strand conformation polymorphism analysis. Genotyping of the identified polymorphisms was performed by restriction fragment length polymorphism. Genotypic association analysis used the Armitage trend test. Haplotype frequency estimation and subsequent testing for differences between cases and controls were performed by using the programs FASTEHPLUS and FAMHAP. RESULTS: We identified 5 novel noncoding nucleotide sequence variants, all of which were typed in our asthmatic, our atopic, and our control population. According to the Armitage trend test, none of the 5 polymorphisms is associated with the phenotype bronchial asthma or atopy. However, multilocus haplotype analysis based on simulations to find out whether the haplotype frequencies differed between cases and controls by using the program FAMHAP yielded a P value of 6.1 x 10(-5) in the asthmatic versus the control population, which is highly significant. Furthermore, we obtained a nominally significant result of P=.0232 for the atopic versus the control population by using FAMHAP. CONCLUSION: These results strongly underscore previous findings that suggest a potential role of this cytokine in allergic diseases. PMID- 15131573 TI - Doxycycline reduces airway inflammation and hyperresponsiveness in a murine model of toluene diisocyanate-induced asthma. AB - BACKGROUND: Toluene diisocyanate (TDI) is a leading cause of occupational asthma. Although considerable controversy remains regarding its pathogenesis, TDI-induced asthma is an inflammatory disease of the airways characterized by airway remodeling caused, at least in part, by an excess of extracellular matrix deposition in the airway wall. Matrix metalloproteinases (MMPs) are major proteolytic enzymes that are involved in extracellular matrix turnover because of their ability to cleave all proteins constituting extracellular matrix. Previous studies have reported that MMP-9 might play a role in chronic airway inflammation and remodeling in asthma. OBJECTIVE: An aim of the current study was to evaluate the effects of MMP-inhibiting antibiotic, doxycycline, and MMP inhibitors on hyperresponsiveness and inflammation of the airways in TDI-induced asthma. METHODS: We used a murine model for TDI-induced asthma to examine the effect of doxycycline or MMP inhibitors on bronchial inflammation and airway hyperresponsiveness. RESULTS: The following typical pathophysiologic features are observed in the lungs of the mice: airway inflammation, airway hyperresponsiveness, and increased expression of MMP-9 mRNA and protein. Administration of doxycycline and MMP inhibitors reduced all of these pathophysiologic findings. In addition, the increased phosphorylated Akt but not Akt protein levels in lung tissues after TDI inhalation were significantly reduced by the administration of doxycycline and MMP inhibitors. CONCLUSION: These findings suggest that doxycycline may reduce airway inflammation and hyperresponsiveness through phosphatidylinositol 3-kinase pathway in a murine model of TDI-induced asthma. PMID- 15131574 TI - Mouse allergen exposure and mouse skin test sensitivity in suburban, middle-class children with asthma. AB - BACKGROUND: Exposure to mouse allergen is prevalent in inner-city homes and is associated with an increased risk of mouse skin test sensitivity in inner-city children with asthma. OBJECTIVE: To determine the distribution of mouse allergen and its relationship to mouse skin test sensitivity in a primarily suburban, middle-class population of asthmatic children. METHODS: Children with asthma, 6 to 17 years old, were recruited from 3 pediatric practices located in counties surrounding the city of Baltimore and from 1 practice located within the city limits. Participants underwent skin prick testing and completed a baseline questionnaire. Their homes were inspected, and settled dust samples were collected for allergen analysis. RESULTS: Two hundred fifty-seven of 335 (76.7%) participants resided outside the city, and 53.7% had annual incomes >$50,000. Mouse allergen was detected in 74.9% of bedrooms, and 13.1% were sensitized to mouse. Lower maternal education (odds ratio [OR], 2.17; 95% CI, 1.28-3.67), city residence (OR, 5.39; 95% CI, 2.23-13.02), and higher bedroom cockroach allergen levels (OR, 9.61; 95% CI, 1.17-79.03) were independent predictors of high bedroom mouse allergen. The risk of mouse skin test sensitivity increased with increasing bedroom Mus m 1 exposure (OR, 1.43; 95% CI, 1.04-1.96, with each increase in quartile), and dog skin test sensitivity was a strong independent predictor of mouse skin test sensitivity (OR, 7.23; 95% CI, 3.03-17.22). CONCLUSION: Mouse allergen exposure is common among suburban, middle-class asthmatic children. Increasing bedroom levels of Mus m 1 and dog skin test sensitivity are risk factors for mouse skin test sensitivity. PMID- 15131575 TI - Immunomodulatory effect of endotoxin on the development of latex allergy. AB - BACKGROUND: Although numerous studies have been conducted delineating the clinical manifestations of latex allergy and characterizing the protein allergens, little is known regarding the natural history of the disease. OBJECTIVE: These studies were undertaken to investigate the immunomodulatory role of inhaled endotoxin on the development of latex-specific IgE-mediated responses to natural rubber latex (NRL) proteins by using a mouse model. METHODS: Female BALB/c mice were exposed to 25 microg of NRL proteins with or without increasing concentrations of endotoxin (50-25,000 EU) through the respiratory tract. Serum antibody levels were evaluated biweekly during the study. After sensitization, mice were challenged with methacholine or NRL proteins, and airway hyperreactivity (AHR) was evaluated with whole-body plethysmography. After NRL challenge, lungs were excised for histopathology, and lung-associated lymph nodes were removed for cytokine mRNA evaluation. RESULTS: When compared with mice exposed to latex alone, mice exposed to latex and endotoxin demonstrated up to 50% lower levels of latex-specific IgE and decreased latex-specific AHR and mucin production. Conversely, these same animals demonstrated increased levels of latex specific serum IgG2a and IgA antibodies and an increase in IFN-gamma and IL-12 mRNA levels in the draining lymph node cells. Concurrent exposure to LPS with nonammoniated latex resulted in increased alveolitis and nonspecific AHR on respiratory challenge with methacholine. CONCLUSION: Coexposure with LPS and allergen decreased latex-specific IgE but augmented nonspecific AHR. These studies demonstrate that endotoxin associated with NRL gloves can modulate the development of allergic responses to NRL proteins. PMID- 15131576 TI - The natural course of atopic dermatitis from birth to age 7 years and the association with asthma. AB - BACKGROUND: Atopic dermatitis (AD) is considered to be one of the first manifestations in the atopic march. However, few prospective studies on AD and its association with childhood asthma exist. OBJECTIVE: The aim of this study was to prospectively investigate the natural course of AD to determine factors influencing its prognosis and to analyze the relationship of AD with childhood asthma. METHODS: The Multicenter Allergy Study, a German birth cohort, followed 1314 children from birth to age 7 years. Physical examinations, parental interviews on atopic symptoms and diagnoses, and determination of specific IgE levels were performed regularly. RESULTS: The cumulative prevalence of AD in the first 2 years of life was 21.5%. Of these children with early AD, 43.2% were in complete remission by age 3 years, 38.3% had an intermittent pattern of disease, and 18.7% had symptoms of AD every year. Severity (adjusted cumulative odds ratio, 5.86; 95% CI, 3.04-11.29) and atopic sensitization (adjusted cumulative odds ratio, 2.76; 95% CI, 1.29-5.91) were major determinants of prognosis. Early wheeze and a specific sensitization pattern were significant predictors for wheezing at school age, irrespective of AD. Early AD without these cofactors constituted no increased risk of subsequent wheeze (adjusted odds ratio, 1.11; 95% CI, 0.56-2.20) or bronchial hyperreactivity. CONCLUSION: AD is a common condition in infancy but disappears around age 3 years in a significant proportion of children. The prognosis is mostly determined by the severity and the presence of atopic sensitization. Early AD is associated with asthma at school age, but in many of these asthmatic children, wheezing manifests before or with the onset of AD. Children with AD and wheeze have a marked loss in lung function, suggesting a distinct phenotype rather than a progressive development from AD to asthma. PMID- 15131577 TI - Cow's milk-specific T-cell reactivity of children with and without persistent cow's milk allergy: key role for IL-10. AB - BACKGROUND: The role of antigen-specific T cells in the mechanism of food allergy or maintenance of tolerance toward an innocuous antigen, such as cow's milk, is not yet fully understood. OBJECTIVE: The cow's milk-specific T-cell response of donors with various allergic backgrounds was investigated. METHODS: Cow's milk specific T-cell clones (TCCs) were generated from the blood of children with persistent cow's milk allergy (CMA) and the blood of cow's milk-tolerant allergic and nonallergic control subjects. The TCCs were characterized by their antigen specific proliferation, cytokine production, and activation status. RESULTS: Cow's milk-specific TCCs of children with persistent CMA were T(H)2 skewed, and the production of IL-4 and IL-13 was significantly correlated with the expression of the activation marker CD25. TCCs of the allergic control subjects were characterized by a high production of IL-10, which was positively correlated with the production of IL-4 and IFN-gamma and with the expression of CD25. TCCs derived from nonallergic control subjects had an attenuated response toward cow's milk in that they did not produce high levels of cytokines nor did they express high levels of surface markers. As in the allergic control subjects, in the nonallergic control subjects IL-10 production was positively correlated with the expression of CD25. CONCLUSION: The activation status of T cells derived from persistent donors with CMA was associated with the production of IL-4 and IL-13, whereas activated TCCs of cow's milk-tolerant control subjects were characterized by the production of IL-10 and, to a lesser extent, IFN-gamma. These findings suggest that activated CD4(+) T cells (characterized by a high CD25 expression) might contribute to the tolerogenic immune response toward an antigen, such as cow's milk, through the production of IL-10. PMID- 15131578 TI - Expression of fractalkine and its receptor, CX3CR1, in atopic dermatitis: possible contribution to skin inflammation. AB - BACKGROUND: Fractalkine (FKN) induces activation and adhesion of leukocytes expressing its receptor, CX(3)CR1. FKN is released from the cell surface through proteolytic cleavage as soluble FKN (sFKN). OBJECTIVE: We sought to assess FKN and CX(3)CR1 expression in the skin, serum sFKN levels, and CX(3)CR1 expression on blood leukocytes in patients with atopic dermatitis (AD). METHODS: FKN and CX(3)CR1 expression in the skin was examined immunohistochemically. mRNA expression of FKN, thymus and activation-regulated chemokine, and macrophage derived chemokine in the skin was assessed by means of real-time RT-PCR. Serum sFKN levels were assessed by using ELISA. Blood leukocytes were stained for CX(3)CR1 by means of flow cytometric analysis. RESULTS: FKN was strongly expressed on endothelial cells in skin lesions of patients with AD and psoriasis but not in normal skin. FKN mRNA levels in AD lesional skin increased to a similar extent to thymus and activation-regulated chemokine and macrophage derived chemokine mRNA levels. CX(3)CR1-expressing cells in the affected skin of patients with AD or psoriasis increased compared with those in normal skin. Serum sFKN levels were increased in patients with AD but not in patients with psoriasis relative to levels in healthy control subjects. Serum sFKN levels were associated with the disease severity and decreased with the improvement of skin lesions in patients with AD. CX(3)CR1(+) cell frequencies and CX(3)CR1 expression levels were decreased in CD8(+) T cells, monocytes, and natural killer cells from patients with AD, but this was not observed in patients with psoriasis. CONCLUSIONS: These results suggest that through functions in both membrane-bound and soluble forms, FKN plays an important role in the trafficking of CX(3)CR1(+) leukocytes during the inflammation caused by AD. PMID- 15131579 TI - FcepsilonRI engagement of Langerhans cell-like dendritic cells and inflammatory dendritic epidermal cell-like dendritic cells induces chemotactic signals and different T-cell phenotypes in vitro. AB - BACKGROUND: Atopic dermatitis (AD) is a biphasic inflammatory skin disease characterized by an initial phase predominated by T(H)2 cytokines, which switches into a second T(H)1-dominated chronic phase. Thus far, the small number of FcepsilonRI-bearing Langerhans cells (LCs) and inflammatory dendritic epidermal cells (IDECs) in the epidermis of patients with AD has hampered a detailed functional analysis and limited our knowledge of these dendritic cells (DCs). OBJECTIVE: We studied FcepsilonRI-mediated mechanisms of LCs and IDECs with the help of a novel in vitro model. METHODS: Langerhans cell-like dendritic cells (LC DCs) and inflammatory dendritic epidermal cell-like dendritic cells (IDEC-DCs) bearing FcepsilonRI have been generated from monocytes of the same atopic donor and compared functionally with LCs and IDECs isolated from the skin of patients with AD. RESULTS: We found that FcepsilonRI-activated LC-DCs release chemotactic signals, and supernatants of FcepsilonRI-activated LC-DCs increase the migratory capacity of precursor cells of IDECs and naive T cells in vitro. FcepsilonRI activated IDEC-DCs produce high amounts of proinflammatory cytokines and chemokines and might thereby amplify the inflammatory immune reaction in patients with AD. Furthermore, FcepsilonRI-activated IDEC-DCs prime naive T cells into IFN gamma-producing T cells and release IL-12 and IL-18, which together might lead to the switch of the initial T(H)2-type immune response into a response of the T(H)1 type in vivo. CONCLUSION: The present study provides evidence that FcepsilonRI activated LC-DCs and IDEC-DCs contribute distinctly to the outcome of T-cell responses in vitro and might have implications for the biphasic nature of AD in vivo. PMID- 15131580 TI - Lymphocytes in Peyer patches regulate clinical tolerance in a murine model of food allergy. AB - BACKGROUND: Food allergy can lead to severe, potentially life-threatening anaphylactic reactions. To generate efficient strategies aimed at an active cure, a better understanding of the pathogenic mechanisms is strongly needed. OBJECTIVE: To investigate T-cell-related mechanisms of food allergy and tolerance to beta-lactoglobulin (BLG) in gut-associated lymphoid structures. METHODS: Beta lactoglobulin-specific IgG1, IgG2a, and IgE in serum from mice anaphylactic to BLG were analyzed by ELISA and compared with those obtained in mice actively tolerized to BLG. The number of Ab-secreting cells in the spleen and in Peyer patches was determined by ELISPOT. The numbers of cytokine-producing cells after antigen-specific activation were measured by the same method. Furthermore, mesenteric lymph node cells and Peyer patches cells were transferred to naive mice, and Ab production as well as Ab-secreting cells were measured. RESULTS: Serum IgG1 and IgE Ab titers as well as IL-4-producing cell numbers were strongly increased in anaphylactic mice. IL-10 was found in Peyer patch cells from tolerant mice after BLG activation but not in anaphylactic mice. Peyer patch cells, in contrast to mesenteric lymph node cells, proliferated weakly in anaphylactic mice after antigen activation, and activation of Peyer patches was partially inhibited by tolerization. CONCLUSIONS: Our data suggest a specific role for lymphocytes in Peyer patches in tolerance to BLG. Low IL-10 production in Peyer patches may favor symptoms of food allergy. PMID- 15131581 TI - Antigen-specific T cell-mediated apoptosis of dendritic cells is impaired in a mouse model of food allergy. AB - BACKGROUND: Dendritic cells (DCs) play a pivotal role in antigen presentation and regulation of immune responses, and strong evidence suggests their involvement in the pathogenesis of allergy. However, hitherto, DC-T-cell cross-talk in relation to IgE-mediated allergic reactions to food has not been investigated. OBJECTIVE: Our aim was to investigate T cell-mediated apoptosis of myeloid DCs from spleen and Peyer's patches of mice with cow's milk (CM) allergy after cognate interaction with antigen (CM)-specific T cells. METHODS: Freshly isolated myeloid CD11c(+/hi)/B220(-) DCs from spleen and Peyer's patches of mice with CM allergy and control mice were cultured with CM-specific T cells in the presence or absence of CM or unrelated antigen as a control. Levels of apoptosis in DCs were evaluated by assessing propidium iodide uptake and annexin V expression by means of flow cytometry. RESULTS: We observed that both systemic and gastrointestinal derived DCs showed an increased resistance to T cell-mediated cell death compared with DCs from control but not allergic donors. Further experiments demonstrated that in both allergic and control mice, T cell-mediated DC apoptosis takes place exclusively in the presence of the specific antigen, is MHC II dependent, and is only partially CD95-CD95 ligand dependent. CONCLUSION: Here we demonstrate, for the first time, that the reciprocal, finely balanced regulation between these 2 cell types, which plays a central role in controlling immune responses, is altered in allergy. We hypothesize that these events are likely to have a profound influence on the genesis and maintenance of adverse reaction to food. PMID- 15131582 TI - Distribution of peanut allergen in the environment. AB - BACKGROUND: Patients with peanut allergy can have serious reactions to very small quantities of peanut allergen and often go to extreme measures to avoid potential contact with this allergen. OBJECTIVE: The purpose of this study was to detect peanut allergen under various environmental conditions and examine the effectiveness of cleaning agents for allergen removal. METHODS: A monoclonal based ELISA for Arachis hypogaea allergen 1 (Ara h 1; range of detection, 30-2000 ng/mL) was used to assess peanut contamination on cafeteria tables and other surfaces in schools, the presence of residual peanut protein after using various cleaning products on hands and tabletops, and airborne peanut allergen during the consumption of several forms of peanut. RESULTS: After hand washing with liquid soap, bar soap, or commercial wipes, Ara h 1 was undetectable. Plain water and antibacterial hand sanitizer left detectable Ara h 1 on 3 of 12 and 6 of 12 hands, respectively. Common household cleaning agents removed peanut allergen from tabletops, except dishwashing liquid, which left Ara h 1 on 4 of 12 tables. Of the 6 area preschools and schools evaluated, Ara h 1 was found on 1 of 13 water fountains, 0 of 22 desks, and 0 of 36 cafeteria tables. Airborne Ara h 1 was undetectable in simulated real-life situations when participants consumed peanut butter, shelled peanuts, and unshelled peanuts. CONCLUSION: The major peanut allergen, Ara h 1, is relatively easily cleaned from hands and tabletops with common cleaning agents and does not appear to be widely distributed in preschools and schools. We were not able to detect airborne allergen in many simulated environments. PMID- 15131583 TI - Should beta-blockers be given to patients with heart disease and peanut-induced anaphylaxis? A decision analysis. AB - BACKGROUND: Beta-blocker therapy postmyocardial infarction is generally recommended because it reduces mortality. However, beta-blockers may increase anaphylaxis mortality in the growing population of patients with peanut-induced anaphylaxis. OBJECTIVE: We sought to assess the risks and benefits of beta blocker therapy among patients with peanut allergy and heart disease. METHODS: We created a Markov model for patients with heart disease at risk for peanut-induced anaphylaxis to compare life expectancy with the following strategies: (1) beta blocker and (2) no beta-blocker. Meta-analysis and a literature review were used to estimate model parameters. We performed sensitivity analysis to explore parameter uncertainty. RESULTS: For peanut-allergic patients who are postmyocardial infarction or who have congestive heart failure, the heart disease benefit of beta-blockers outweighs the increased likelihood of dying from anaphylaxis, increasing life expectancy by 9.4 and 17.4 months, respectively. Beta-blocker was preferred unless (1) the annual rate of moderate to severe anaphylaxis exceeded 6.0% for postmyocardial infarction and 15% for congestive heart failure patients; (2) beta-blocker therapy increased the incidence of moderate to severe anaphylaxis >2.5-fold for postmyocardial infarction and >5.8 fold for congestive heart failure patients; (3) anaphylaxis case fatality exceeded 6.5% postmyocardial infarction; or (4) beta-blocker therapy increased anaphylaxis case fatality >25-fold postmyocardial infarction. CONCLUSION: Our results suggest that for patients postmyocardial infarction or with congestive heart failure who are at risk for peanut-induced anaphylaxis, beta-blocker use should still improve survival. However, the epidemiology of anaphylaxis and effects of beta-blocker therapy on anaphylaxis incidence and mortality require further study. PMID- 15131584 TI - How do we avoid developing allergy: modifications of the TH2 response from a B cell perspective. AB - The synthesis of IgE by B cells occurs at a low rate compared with that of other antibodies, even in allergic subjects. One rate-limiting step is the class switch, by which B lymphocytes switch to produce immunoglobulin epsilon heavy chains rather than micro or gamma heavy chains. We propose an additional rate limiting step: survival of the B lymphocyte after the switch to IgE. The hypothesis we present here is that the survival of the IgE-switched B cell is compromised, particularly in an active germinal center. Antigenic stimulation in the absence of a danger signal fails to induce a mature germinal center, which allows IgE-switched B cells to escape and mature into plasma cells. Antigenic stimulation in the presence of a danger signal (or under nonhygienic conditions) induces germinal centers, which eliminate IgE-switched B cells. Thus the essence of an allergen is antigenic stimulation in the absence of conditions that generate mature germinal centers. Because germinal centers are important for the generation of B-cell memory, the IgE immune response is characteristically poor in memory (but might be long lasting because of the generation of long-lived plasma cells). In addition to this direct route to IgE, typical for atopic sensitization, another type of T(H)2 response exists. On chronic allergen exposure with the concomitant induction of germinal centers, IgG4-switched B memory cells are induced that are slow to differentiate into plasma cells. These IgG4-switched B memory cells might occasionally undergo a secondary switch to IgE. PMID- 15131585 TI - Sustained T-bet expression confers polarized human TH2 cells with TH1-like cytokine production and migratory capacities. AB - BACKGROUND: The transcription factor T-bet mediates IFN-gamma production by T(H)1 cells and suppresses T(H)2 cytokine production when ectopically expressed in polarized murine T(H)2 cells. Thus T-bet-mediated inhibition of T(H)2 cytokine production might be beneficial for the treatment of allergic diseases like asthma or atopic dermatitis. OBJECTIVE: We sought to investigate the effects of ectopic T-bet expression in highly polarized human T(H)2 cells obtained from skin biopsy specimens of patients with atopic dermatitis. METHODS: The cytokine production of T(H)2 cells retrovirally transfected with a vector expressing human T-bet was determined by means of intracellular FACS staining and ELISA. The effects of T bet transfection were analyzed at the mRNA level by means of real-time PCR and DNA microarrays and confirmed by using functional chemokine response assays. RESULTS: Transfection of T-bet into T(H)2 cells induced high levels of IFN-gamma and suppressed IL-5, but IL-2 and IL-4 production remained unchanged. T-bet transfection also induced IL-12Rbeta2 and CXCR3 expression on human T(H)2 cells, whereas the IL-18 receptor was only induced as a consequence of T-bet-mediated increased responsiveness to IL-12. Furthermore, sustained T-bet expression in human T(H)2 cells induced IL-2 production and decreased the secretion of IL-4. In addition, the chemokine receptor repertoire of these cells was changed toward a T(H)1-like profile. CONCLUSION: The combined switch in cytokine pattern and migratory potential of highly polarized human T(H)2 cells mediated by T-bet might provide an additional advantage for the treatment of allergic diseases. PMID- 15131586 TI - Immunopathology of food allergy. PMID- 15131588 TI - The different apoptotic potential of the p53 codon 72 alleles increases with age and modulates in vivo ischaemia-induced cell death. AB - A common arginine to proline polymorphism is harboured at codon 72 of the human p53 gene. In this investigation, we found that fibroblasts and lymphocytes isolated from arginine allele homozygote centenarians and sexagenarians (Arg+) undergo an oxidative-stress-induced apoptosis at a higher extent than cells obtained from proline allele carriers (Pro+). At variance, the difference in apoptosis susceptibility between Arg+ and Pro+ is not significant when cells from 30-year-old people are studied. Further, we found that Arg+ and Pro+ cells from centenarians differ in the constitutive levels of p53 protein and p53/MDM2 complex, as well as in the levels of oxidative stress-induced p53/Bcl-xL complex and mitochondria-localised p53. Consistently, all these differences are less evident in cells from 30-year-old people. Finally, we investigated the in vivo functional relevance of the p53 codon 72 genotype in a group of old patients (66 99 years of age) affected by acute myocardial ischaemia, a clinical condition in which in vivo cell death occurs. We found that Arg+ patients show increased levels of Troponin I and CK-MB, two serum markers that correlate with the extent of the ischaemic damage in comparison to Pro+ patients. In conclusion, these data suggest that p53 codon 72 polymorphism contributes to a genetically determined variability in apoptotic susceptibility among old people, which has a potentially relevant role in the context of an age-related pathologic condition, such as myocardial ischaemia. PMID- 15131589 TI - Developmental stage-dependent self-regulation of embryonic cortical precursor cell survival and differentiation by leukemia inhibitory factor. AB - Cortical precursor cells secrete soluble factors for their own survival and self renewal. We show here that neural precursor cells isolated from embryonic rat cortices abundantly secrete leukemia inhibitory factor (LIF) and express its receptor components, gp130 and LIF receptor. LIF signaling is responsible for cortical precursor cell survival. As described previously, LIF caused astrocytic differentiation of cultured embryonic cortical precursor cells. LIF-mediated survival and astrocytic differentiation of cortical precursor cells were differentially regulated, depending on the developmental ages of embryos from which cortical precursors were isolated. LIF did not enhance the survival of cortical precursor cells isolated from later embryos (embryonic day 16, E16). Moreover, LIF-mediated astrocytic differentiation was not observed in early (E12) cortical precursors. Inhibition studies revealed that Janus-activated kinase/signal transducer and activator of transcription and phosphatidylinositol 3 kinase/Akt pathways participate in both the LIF-mediated effects. However, mitogen-activated protein kinase, another signal pathway activated by LIF, was specifically responsible for astrocytic differentiation. These findings collectively indicate that precursor cells self-regulate the sequential processes of brain development, such as early maintenance of the precursor cell population and later differentiation into astrocytes, via common LIF signaling. PMID- 15131590 TI - Endoplasmic reticulum stress response is involved in nonsteroidal anti inflammatory drug-induced apoptosis. AB - Apoptosis induced by nonsteroidal anti-inflammatory drugs (NSAIDs) is involved not only in the production of NSAID-induced gastric lesions but also in the antitumor activity of these drugs. The endoplasmic reticulum (ER) stress response is a cellular mechanism that aids in protecting the ER against ER stressors and is involved in ER stressor-induced apoptosis. Here, we examine the relationship between this response and NSAID-induced apoptosis in cultured guinea-pig gastric mucosal cells. Exposure of cells to indomethacin, a commonly used NSAID, induced GRP78 as well as CHOP, a transcription factor involved in apoptosis. Three factors that positively regulate CHOP expression (ATF6, ATF4 and XBP-1) were activated and/or induced by indomethacin. NSAIDs other than indomethacin (diclofenac, ibuprofen and celecoxib) also induced CHOP. Monitoring of the transcriptional activities of ATF6 and CHOP by luciferase assay revealed that both were stimulated in the presence of indomethacin. Furthermore, indomethacin induced apoptosis was suppressed in cultured guinea-pig gastric mucosal cells by expression of the dominant-negative form of CHOP, or in peritoneal macrophages from CHOP-deficient mice. These results suggest that ER stress response-related proteins, particularly CHOP, are involved in NSAID-induced apoptosis. PMID- 15131591 TI - Molecular mechanisms of TNF-alpha-induced ceramide formation in human glioma cells: P53-mediated oxidant stress-dependent and -independent pathways. AB - The present study was designed to examine the roles of p53, reactive oxygen species (ROS), and ceramide, and to determine their mutual relationships during tumor necrosis factor (TNF)-alpha-induced apoptosis of human glioma cells. In cells possessing wild-type p53, TNF-alpha stimulated ceramide formation via the activation of both neutral and acid sphingomyelinases (SMases), accompanied by superoxide anion (O2-*) production, and induced mitochondrial depolarization and cytochrome c release, whereas p53-deficient cells were partially resistant to TNF alpha and lacked O2-* generation and neutral SMase activation. Restoration of functional p53 sensitized glioma cells expressing mutant p53 to TNF-alpha by accumulation of O2-*. z-IETD-fmk (benzyloxycarbonyl-Ile-Glu-Thr-Asp fluoromethyl ketone), but not z-DEVD-fmk (benzyloxycarbonyl-Asp-Glu-Val-Asp fluoromethyl ketone), blocked TNF-alpha-induced ceramide formation through both SMases as well as O2-* generation. Caspase-8 was processed by TNF-alpha regardless of p53 status of cells or the presence of antioxidants. Two separate signaling cascades, p53 mediated ROS-dependent and -independent pathways, both of which are initiated by caspase-8 activation, thus contribute to ceramide formation in TNF-alpha-induced apoptosis of human glioma cells. PMID- 15131592 TI - Mutational analysis of P-glycoprotein: suppression of caspase activation in the absence of ATP-dependent drug efflux. AB - P-glycoprotein (P-gp) can induce multidrug resistance (MDR) through the ATP dependent efflux of chemotherapeutic agents. We have previously shown that P-gp can inhibit nondrug apoptotic stimuli by suppressing the activation of caspases. To determine if this additional activity is functionally linked to ATP hydrolysis, we expressed wild-type and ATPase-mutant P-gp and showed that cells expressing mutant P-gp could not efflux chemotherapeutic drugs but remained relatively resistant to apoptosis. CEM lymphoma cells expressing mutant P-gp treated with vincristine showed a decrease in the fraction of cells with apoptotic morphology, cytochrome c release from the mitochondria and suppression of caspase activation, yet still accumulated in mitosis and showed a loss of clonogenic potential. The loss of clonogenicity in vincristine-treated cells expressing mutant P-gp was associated with accumulation of cells in mitosis and the presence of multinucleated cells consistent with mitotic catastrophe. The antiapoptotic effect of mutant P-gp was not affected by antibodies that inhibit the efflux function of the protein. These data are consistent with a dual activity model for P-gp-induced MDR involving both ATPase-dependent drug efflux and ATPase-independent inhibition of apoptosis. The structure-function analyses described herein provide novel insight into the mechanisms of action of P-gp in mediating MDR. PMID- 15131593 TI - Finding the balance. PMID- 15131596 TI - Capitation plans. PMID- 15131598 TI - Glass ionomer cements. PMID- 15131600 TI - Postoperative antibiotics. PMID- 15131601 TI - Admission statistics. PMID- 15131602 TI - Questionnaire research. PMID- 15131604 TI - Appropriate insurance. PMID- 15131605 TI - Hygienist training. PMID- 15131606 TI - Oral health educators. PMID- 15131615 TI - Research in primary dental care. Part 1: Setting the scene. AB - There is a growing trend for research to be conducted in primary health settings, to the benefit of patients and healthcare workers. This article provides an introduction and overview of a series which aims to facilitate research in general dental practice and to encourage general dental practitioners and other members of the dental team to become involved in these studies. The benefits of undertaking research are outlined, as well as barriers to becoming involved in such research. It is argued that many of these barriers can be overcome by encouraging dental practitioners to collaborate with established researchers in a variety of settings. PMID- 15131616 TI - Miconazole oral gel and drug interactions. AB - Miconazole oral gel is frequently prescribed for the treatment of oral Candidal infections. Its ability to be systemically absorbed and interact with other drugs has previously been recorded but is not universally known. As a reminder, a further case of derangement of anticoagulation following concomitant use of warfarin and miconazole is reported. Other potential drug interactions of miconazole and fluconazole are highlighted. PMID- 15131625 TI - In vitro and in vivo assessment of a glass slow fluoride releasing device: a pilot study. AB - AIMS: The aims were to evaluate a) whether a slow release fluoride-glass pellet (SFG) would be retained in the mouth and release fluoride (F) over a long period of time, b) what concentrations of F in the glass would provide ideal intra-oral saliva F concentrations and c) whether an SFG would affect blood plasma concentrations of F after swallowing compared with ingestion of a commercial NaF tablet. METHODS: a) A prototype SFG was attached to a maxillary molar of a volunteer. Baseline saliva F concentrations were measured prior to glass placement, daily in week one; one day a week for weeks two to three and then one day a month up to 18 months. Four subjects had the SFG for six months with saliva F concentration assessments at periodic intervals. b) SFGs containing F at 13.3%, 18.3% and 21.9%, and an improved solubility, were tested using three volunteers and saliva F concentrations measured. c) Five volunteers each swallowed either a SFG or a NaF tablet. Blood plasma samples were taken at baseline and F measured at time intervals of 2.5, 5, 10, 20, 30, 45, 60, 90 and 120 mins post-ingestion. RESULTS: a) The prototype SFG were successfully retained and released F into saliva; mean concentrations of 0.035 mg L(-1) were achieved lasting for over 18 months. Overall saliva F concentrations were approximately doubled. Analysis of the pellet at the end of use showed it still contained some F possibly indicating a recharging effect. b) The 13.3% F concentration SFG produced significantly higher saliva F levels than the two other concentrations tested. The two higher concentration F glasses contained aluminium as part of the formulation of the glass structure, which is known to bind F whereas the 13.3% glass contained F alone. c) While blood plasma F levels increased after ingestion of the NaF tablet there was no increase in F when the SFG was swallowed. CONCLUSION: A slow release F containing glass device showed promise as a means to enhance intra-oral F saliva concentration. PMID- 15131626 TI - Changes in water intake of Northumbrian adolescents 1980 to 2000. AB - OBJECTIVES: To determine: total water intake in young English adolescents; the relative importance of sources of water intake; and changes in water intake and sources of water between 1980 and 2000. DESIGN: A cross-sectional observational study of the diets of 11-12-year-old children attending seven schools in south Northumberland, UK. The information obtained was compared with results from a similar survey carried out 20 years previously. METHODS: All children attending these schools were invited to participate. They completed a three-day diet diary with an interview on the fourth day, on two occasions during the school year. Standard UK food composition tables were used and water intake from various sources calculated. Anthropometric and social class information was obtained. RESULTS: Four hundred and twenty-four children completed all aspects of the study (64% of those eligible). The mean total water intake was 1,130 g d(-1), approximately the same as that recorded 20 years before. Water intake in relation to energy intake: water intake was 139 g MJ(-1) in boys and 143 g MJ(-1) in girls. Sixty-five per cent of water came from drinks and 35% from foods; very similar to proportions 20 years before. The sources of water in drinks had changed considerably, with a marked increase in consumption of soft drinks (especially carbonated drinks) and a decrease in consumption of hot drinks and milk. There were a few differences between sexes but little difference between social groups. CONCLUSIONS: Total water intake was similar to that recorded 20 years previously and lower than intakes reported in other countries. Changes in the sources of water meant that less water consumed was likely to come from the tap in the house and more from drinks made elsewhere, than 20 years previously. These changes have implications for estimating fluoride intake in fluoridated areas. PMID- 15131627 TI - Bicycle helmets--does the dental profession have a role in promoting their use? AB - OBJECTIVES: To review the available literature regarding the: epidemiology of bicycle related head injuries; consequences of head injuries; rates of cycle helmet use; impact of educational campaigns and legislation on usage rates; effectiveness of cycle helmets in protecting against head and facial injuries; arguments against the compulsory use of bicycle helmets. DATA SOURCES: A computerised Medline search was conducted using the keywords: head injury, facial injury, bicycle helmets, accidents. DATA SELECTION: All available information was considered. DATA SYNTHESIS: Data was collated manually. CONCLUSIONS: The wearing of bicycle helmets contributes significantly to the prevention of head injuries (HI) and traumatic brain injury (TBI), particularly in children and adolescents. There is evidence to support the role of cycle helmets in the prevention of injuries to the middle third of the face and some dental injuries. There is a case for the implementation of legislation accompanied by educational campaigns to increase significantly the use of cycle helmets. The dental profession could: play an active role in promoting cycle helmet use; support calls for the compulsory wearing of cycling helmets, particularly for children; press for modification of helmet design and standards to increase protection of the face. PMID- 15131628 TI - An investigation into the use of bitewing radiography in children in Greater Glasgow. AB - AIM: In recent years there have been a number of reviews and guidelines published with respect to the diagnosis and management of caries in children. Bitewing radiography remains the recommended method of choice for caries diagnosis in most circumstances. The aims of the study were to investigate the usage of bitewing radiography by general dental practitioners (GDPs) in Greater Glasgow, for the diagnosis of caries in children and to assess the usefulness of the technique as perceived by these dental practitioners. DESIGN: A questionnaire for self completion was sent to all GDPs with an NHS list number in the Greater Glasgow area. RESULTS: An 80% response rate was obtained with 303 GDPs responding. Less than half the dentists (44%) indicated that they always carried out caries risk assessments for all children, but 71% reported doing this for 6 and 7-year-old children. Approximately 60% of dentists stated they had read the recent radiation and caries management guidelines. Only 72% stated that they used radiography as a caries diagnostic tool in children, with 12 GDPs (4%) indicating they would never consider using bitewing radiography in children. Only 17% would consider taking bitewing radiographs in children under 6 years, and the majority of dentists (61%) reported that they would first consider taking radiographs in the 6-11-year old age group. However, only a low proportion of children in this age band had dental x-rays taken. Bitewing radiography was considered to be more important for 12-year-olds than for 6-year-olds. Recommended methods for decreasing radiation exposure such as rectangular collimation and film holders were not being used universally, with 41% and 58% respectively using these devices. CONCLUSIONS: A significant proportion of dentists in Greater Glasgow are not complying with recent guidelines and recommendations and it would appear that the value of bitewing radiography as a diagnostic tool in children is not being fully exploited. PMID- 15131629 TI - The future of teaching of complete denture construction to undergraduates in the UK: is a replacement denture technique the answer? AB - Due to changes in the amount of curriculum time available for teaching complete denture construction to undergraduate students, course content requires reconsideration and possible modification. The idea that a replica complete denture technique may offer some advantages is explored. A review of the relevant literature fails to support many of the claimed benefits of this technique such as relative ease of adaptation by the elderly to new dentures. This article suggests that the replica denture technique should be considered as no more than an alternative method of making dentures. The relative demerits of the technique are discussed. A modification of the replica block technique to include important aspects of conventional complete denture courses is described. It is suggested that, because less clinical time is required, it may be a suitable way of introducing students to complete denture construction and educating them to a level which would encourage some to take further postgraduate study and would allow all graduates to treat routine cases or diagnose, treatment plan and refer to clinical dental technicians. PMID- 15131649 TI - The Bayesian revolution in genetics. PMID- 15131650 TI - Chemogenomics: an emerging strategy for rapid target and drug discovery. PMID- 15131651 TI - Applied bioinformatics for the identification of regulatory elements. PMID- 15131652 TI - Adaptive evolution and explosive speciation: the cichlid fish model. PMID- 15131653 TI - The evolutionary dynamics of eukaryotic gene order. PMID- 15131654 TI - RNA regulation: a new genetics? PMID- 15131655 TI - Abstracts of the 18th European Histocompatability Conference, 8-11 May 2004, Sofia, Bulgaria. PMID- 15131672 TI - Uveitis and the menstrual cycle. AB - PURPOSE: To establish whether there is a variation in the incidence of onset of acute anterior uveitis (AAU) in women during the phases of the menstrual cycle. METHODS: Prospective open study in women attending the Acute Referral Centre with a first or recurrent attack of AAU. RESULTS: There was a significant increase in the incidence of AAU during the late stages of the menstrual cycle. CONCLUSIONS: The onset of AAU is partially dependent on the levels of either oestrogen or progesterone, or both. The withdrawal of the proven anti-inflammatory effects of these hormones may provoke the onset of uveitis. PMID- 15131673 TI - Polypoidal choroidal vasculopathy and exudative age-related macular degeneration in Greek population. AB - PURPOSE: To study the prevalence, the clinical features, and the visual prognosis without treatment of polypoidal choroidal vasculopathy (PCV) in a large series of Greek patients presenting with exudative maculopathy. METHODS: The medical records, photographs,as well as fluorescein and indocyanine green(ICG) angiograms of a series of 268 consecutive elderly white Greek patients, who were originally diagnosed as having exudative age-related macular degeneration (AMD) were reviewed retrospectively. RESULTS: In all, 22 of the 268 (8.2%) patients initially suspected of having AMD were ultimately diagnosed with PCV. In 15 of the 22(68.2%) patients with PCV, the polypoidal lesions were located in the peripapillary area. Large soft drusen were present in only two fellow eyes of the 10 (20%) patients with unilateral PCV compared with 120 fellow eyes of the 148 (81.1%) patients with unilateral AMD. At the last examination, 11 of the 22(50%) patients with PCV and 120 of the 246(48.8%) patients with AMD presented a visual acuity of less than 6/60 in at least one eye due to scar formation in the macula. CONCLUSIONS: PCV is not an infrequent disease in Greece. A measurable number of Greek patients with findings suggestive of exudative AMD will instead have PCV. ICG angiography is important in differentiating between these two clinical entities. In Greeks, polypoidal lesions are predominantly peripapillary and are not usually associated with macular drusen in the fellow eye. PCV and exudative AMD do not differ significantly in terms of their natural course and visual prognosis in Greek patients. PMID- 15131674 TI - Hyperhomocysteinemia in patients with diabetes mellitus with and without diabetic retinopathy. AB - OBJECTIVE: To evaluate the prevalence of hyperhomocysteinaemia in diabetic patients with no diabetic retinopathy (no DR), with non-proliferative diabetic retinopathy (NPDR) and with proliferative diabetic retinopathy (PDR). RESEARCH DESIGN AND METHODS: This prospective, case-control study, included 179 diabetic patients and 156 age-matched controls with no diabetes and no history of ocular disease, who were undergoing routine physical checkups. Plasma homocysteine levels of all study participants were measured using high-performance liquid chromatography (HPLC). Hyperhomocysteinaemia was defined when homocysteine levels were higher than 15 micromol/l. RESULTS: The mean plasma homocysteine level was 11.75+-0.24 in the control group,13.46+0.74 in the no DR group, 14.56 + 0.64 in the NPDR group and 15.86 + 1.34 in the PDR group. Mean homocysteine levels were significantly elevated in the NPDR and PDR groups compared to the control group(P = 0.001 and <0.0001, respectively). The prevalence of hyperhomocysteinaemia was also higher in the NPDR and PDR groups compared to the control group (P = 0.032 and 0.011, respectively). No statistically significant difference was found between the no DR and the control group. CONCLUSIONS: Our findings suggest that hyperhomocysteinaemia may be associated with diabetic retinopathy and partially explain the increased risk of microvascular angiopathy occurring in these patients. PMID- 15131675 TI - A single transcutaneous injection with Botox for dysthyroid lid retraction. AB - PURPOSE: To evaluate the safety and efficacy of injections with botulinum toxin type A (BTTA,Botox), given transcutaneously, in the treatment of upper lid retraction associated with thyroid eye disease (TED). METHODS: A total of 15 patients (21 eyes) with a stable (TED) condition, and a euthyroid state, were enrolled into the study. There were 12 females and three males from ages 23 to 52 years. A single injection, at the centrally superior tarsal border transcutaneously, aiming at the levator aponeurosis and Muller muscle, was administered into each eyelid with 5-6 U of Botox. All patients were followed regularly for 4-6 months. Any complications, such as ptosis, diplopia, pain,or lid ecchymosis were recorded. RESULTS: All patients, except one, experienced much reduction of palpebral fissure. The mean difference of MRD1 between pre- and postinjections of Botox at the first week was -3.1 mm, and the effect remained, at least, for 2 months. There were temporary complications of ptosis in three patients and vertical diplopia in two patients, lasting 3-4 weeks. CONCLUSIONS: A single transcutaneous injection with Botox for the treatment of thyroid lid retraction is safe and effective. Some minor complications may occur, such as ptosis and diplopia; however, it may offer an alternative and temporary method for patients with dysthyroid lid retraction, who are waiting for a staged operation of either an orbital decompression or a strabismus surgery or both. PMID- 15131676 TI - Eye nutrient products for age-related macular degeneration: what do they contain? AB - BACKGROUND AND PURPOSE: Patients are increasingly well informed about the availability of antioxidant products and the claims made for their benefits in age-related macular degeneration (AMD). Consequently, their use is becoming widespread. The purpose of this study is to conduct a survey of the commonly encountered products, and to compare their ingredients with the current Age Related Eye Disease Study (AREDS) recommendations. METHODS: A search was undertaken for products sold as 'eye nutrients' at local pharmacies and health food shops, and for products advertised via the Internet. Information about these products was collated and analysed. RESULTS: We identified 22 eye nutrient products. Analysis of their constituents showed that, although over 75% contained all the constituents used in AREDS, only two matched the dosage profiles recommended in the study. CONCLUSION: The authors draw no conclusion on the efficacy of nutritional supplements in the prevention of AMD. In order to advise their patients, ophthalmologists should be familiar with these products. The compiled list in this paper should provide a useful reference for them. PMID- 15131677 TI - Fluorescein angiography-guided indocyanine green angiography for the detection of feeder vessels in subfoveal choroidal neovascularization. AB - AIMS: To compare the newly devised fluorescein angiography (FA) - guided indocyanine green angiography (ICGA) with conventional ICGA for detecting feeder vessels in subfoveal choroidal neovascularization (CNV). METHODS: The detection of feeder vessels was attempted in 86 AMD patients with subfoveal CNV: 44 (age 70.4+/-4.5 years) underwent conventional ICGA (control group) and 42 (age 70.9+/ 4.0 years) underwent an FA-guided ICGA (FA-guided group) using a double-detector scanning laser ophthalmoscope (SLO). In the control group, indocyanine green (ICG) was injected simultaneously with fluorescein. The patients were instructed to gaze forward localizing the fovea at the centre. In the FA-guided group, fluorescein sodium was injected a few minutes prior to ICG, and the patients were instructed to fixate in the appropriate direction by referring to the ongoing FA on the monitoring screen. In both groups, a 20 degrees visual angle was used to capture good images of feeder vessels in ICGA and, in case ICGA missed the first images of the entire CNV filling, an additional injection of ICG was given in the late phase to record the choroidal filling again. The overall detection rate, single-injection detection rate, double-injection rate and examination time were analysed using Fisher's direct exact probability test or Mann-Whitney's U-test. RESULTS: There was no significant difference in the overall detection of feeder vessels between the two groups (50% in the control group and 52.3% in the FA guided group; P=0.49 with Fisher's direct exact probability test). However, in the FA-guided group, the single injection detection rate was significantly higher (45.1 and 15.9%, respectively; P<0.001 with Fisher's direct exact probability test); significantly less double injections were required (7.1 and 50%, respectively; P=0.003 with Fisher's direct exact probability test); and significantly shorter examination times were needed (9.6+/-3.7 and 14.1+/-6.8 min, respectively; P=0.02 with Mann-Whitney's U-test). CONCLUSION: FA-guided ICGA is effective for detecting feeder vessels of subfoveal CNV, minimizing the amount of ICG injected and the examination time compared to conventional ICGA. PMID- 15131678 TI - Can a public health package on glaucoma reach its target population? AB - PURPOSE: A pilot study to assess how successful a newspaper advertisement and a radio interview about glaucoma are at reaching their target population. METHODS: The health intervention comprised two components: an interview on local radio and an advertisement in the local paper. Our target population were residents aged 45 years and above in either Southall (West London) or the Isle of Wight (IOW). A questionnaire was developed to be carried out pre- and post-intervention. The data from both locations pre and post were coded and cleaned. Tests of significance were carried out to assess statistical significance for differences in proportion, with tests for trend used where appropriate. All statistical analyses were carried out using Stata7. RESULTS: Overall, the proportion who had heard of glaucoma increased from 54% before the intervention to 60% after (chi(2) = 3.7, P = 0.055). The proportion who had heard of the disease increased by 13% (chi(2) = 8.76, P = 0.003) in Southall and by 8% (chi(2) = 5.02, P = 0.025) on the IOW. The proportion reporting seeing the advert increased significantly in both areas with greater effect in Southall. Those reporting hearing the radio interview only increased in Southall. On the IOW, females were more knowledgeable and responded more positively to the intervention. This differed in Southall where males tended to be the positive responders. Conclusion In both areas a significant effect on those having heard of glaucoma was found. This could be attributed to both the advert and interview in Southall but would appear to be attributable to the newspaper advertisement alone on the IOW. PMID- 15131679 TI - Glaucoma case finding: a cluster-randomised intervention trial. AB - PURPOSE: To assess the effect of an intervention comprising training in optic disc assessment, explicit referral criteria and ophthalmologist feedback on referred patients, on the number of optometrist referrals for suspected glaucoma seen at a referral site and the positive predictive value of those referrals. METHODS: Optometric practices routinely referring to the Ealing Hospital Eye Clinic were randomly divided into two groups taking into consideration those practices, which shared an optometrist (a cluster) and the number of optometrist days worked per week. One group of practices acted as controls, while the other practices were invited to receive the intervention. Data on 397 new patients referred and presenting to Ealing Hospital with suspected glaucoma were collected over a 20-month period. The data on patients who had failed to attend their appointment were collected over 7 months of this period. The number of referrals seen, the positive predictive value of those referrals, and the attendance rate were calculated. Optometrist's opinions of the intervention were assessed qualitatively. Data relating to optometrist compliance with the intervention were also collected. RESULTS: The number of glaucoma referrals presenting to Ealing Hospital from the intervention practices was almost double that from the control practices (210 vs 119). When cluster randomisation, the number of optometrist days per cluster and the number of assessed referrals in the preintervention period are taken into consideration, it is estimated that the intervention is associated with a 52% increase in the number of referrals reaching Ealing Hospital. However, the design effect resulting from the cluster randomisation was unexpectedly high (of the order of 13-14)and so the confidence intervals around the estimate of 52% are very wide (95% c.i. 35% decrease to 253% increase, P = 0.34). There was no evidence of an association between optometrist compliance with the intervention and the number of referrals seen at Ealing Hospital. The positive predictive value (PPV)of referrals was similar for the intervention(0.49 (95% c.i. 0.42, 0.55)) and control groups(0.46 (95% c.i. 0.33, 0.60)). Optometrist opinions of the intervention were largely favourable. All expressed a willingness to participate in future programmes. CONCLUSION: A large difference in the number of referrals between the practice groups was observed. Since the PPV of referral was unchanged, the potential impact of the intervention in terms of numbers of new cases of glaucoma detected in the community is substantial. However, because of its large design effect, this trial does not provide conclusive evidence of an impact of the intervention on referral numbers. A considerably larger trial will be required to produce conclusive evidence of an effect. PMID- 15131680 TI - Prevalence of glaucoma in an African population. AB - PURPOSE: To determine the prevalence of primary open-angle glaucoma (POAG) in a Ghanaian population aged 30 years and above and to describe any ethnic variations in glaucoma prevalence in this population. METHOD: A cross-sectional prevalence survey for POAG was carried out on residents of 30 years and above in the Akwapim South district of Ghana. The principal investigator examined all glaucoma suspects and those diagnosed as glaucoma in the initial screening to determine all definite cases of POAG. All cases had intraocular pressure measurements. Glaucoma cases and suspects and some normal subjects had fundus biomicroscopy with 78 D Volk lens, and Humphrey FDT N-30 visual fields plotted. RESULTS: A total of 1843 people aged 30 years were screened from the population. The standardized age-specific prevalence was 7.7% (30 years and above) and 8.5% (40 years and above). The best-fit trend line for prevalence/age relationship was exponential. No gender or ethnic difference in prevalence was found. CONCLUSIONS: The prevalence of POAG in this population is high and comparable to those in black populations in Barbados and St. Lucia PMID- 15131681 TI - Selective vs argon laser trabeculoplasty: hypotensive efficacy, anterior chamber inflammation, and postoperative pain. AB - PURPOSE: To compare selective laser trabeculoplasty (SLT) with conventional argon laser trabeculoplasty (ALT) in terms of hypotensive efficacy, anterior chamber inflammation, and pain reported by the patients treated. METHODS: A prospective study performed on 40 consecutive patients. Group I (n = 20): SLT 180 degrees. Group II (n = 20): ALT 180 degrees. Intraocular pressure, flare (Laser-Flare Meter, Kowa FM-500, Japan), and pain (Visual Analogue Scale) were measured before treatment and 1 h, 24 h,1 week, and 1, 3 and 6 months after treatment. Statistically significant differences were determined by an independent-sample Student's t-test. RESULTS: At 6 months after treatment, pressure reduction was similar in both groups: SLT 22.2% (range 0-36.3%) and ALT 19.5% (range 0-30.2%), P= 0.741. The energy released during treatment was significantly lower in SLT (48.3 SD 7.4 mJ) than in ALT (4321 SD 241.7 mJ), P < 0.001. At 1 h after treatment,anterior chamber flare was also lower in SLT(13.3 SD 6.3 vs 20.7 SD 7.4 photons/ms),P = 0.003. Pain reported by the patients during the treatment was significantly lower in SLT(2.0 SD 0.7 vs 4.3 SD 1.3), P<0.001. CONCLUSIONS: The hypotensive efficacy of both lasers at the end of follow-up was similar. The energy released during treatment and inflammation produced in the anterior chamber in the immediate postoperative period were significantly lower for SLT. The SLT procedure was better tolerated, producing less discomfort during treatment than conventional trabeculoplasty with argon. PMID- 15131682 TI - The Amsler chart is of doubtful value in retinal screening for early laser therapy of subretinal membranes. The West London Survey. AB - PURPOSE: Validating the current protocol of Amsler chart grid surveillance for the early detection of subretinal neovascular membrane(SRN) in age-related macular degeneration(AMD), and investigating its value in facilitating early laser therapy. METHODS: A retrospective pilot study. SETTING: Central London eye hospital with dedicated 24-h ophthalmic casualty serving West and West-central London. PARTICIPANTS: 100 consecutive AMD patients who attended casualty with vision loss fulfilling the following criteria: patients had received and been instructed in the use of Amsler charts according to the unit's dispensation protocol,fluorescein angiography which confirmed new SRN. Patients presented over 20 months. Outcome measures were detection of SRN by the Amsler chart, and laser treatment of SRN. RESULTS: The Amsler chart surveillance protocol had detected SRN in 29 of the 100 patients. The surveillance protocol detected less than 30% of the specific patients who subsequently underwent laser treatment. A statistically significant difference was seen on comparing the ages of patients in whom the screening protocol was successful versus those in whom it was unsuccessful (student's t-test,P<3.2 x 103). Younger patients were more likely to be detected using the Amsler chart. A one-tailed Z2 test approached, but did not achieve, statistical significance (Z2 = 1.057,P <0.3) suggesting that patients who have already lost vision to SRN in one eye might not be more likely to be detected using the surveillance protocol than patients in whom SRN was affecting their first eye. In all, 38% of surveillance responders went on to receive laser therapy, compared with 37% of surveillance nonresponders. CONCLUSIONS: The current Amsler chart surveillance protocol is suboptimal for detecting SRN in AMD, and a proportion of cases suitable for early laser therapy may be missing rapid detection. The results are especially important since recent advances in laser therapy for SRN require early detection for optimal effectiveness. PMID- 15131683 TI - Knowledge, self-help and socioeconomic factors in South Asian and Caucasian diabetic patients. AB - AIMS: We carried out a survey of important nonclinical issues including awareness and self-management of diabetes on a group of South Asian and Caucasian patients attending diabetic clinics within a set period. METHODS: A structured questionnaire examined various issues including demographics, perceived knowledge and awareness of diabetes, perceived self-help/support, and psycho-social factors. A total of 500 patients (268 South Asians and 232 Caucasian) took part. RESULTS: Univariate analysis showed significant differences (P<0.05) with various issues including a lower perceived awareness of diabetes and its complications in South Asians, and of the nutritional content of their diet. Asians also appeared to be less worried in the event of missed clinical appointments and if treatment was not strictly adhered to. CONCLUSIONS: The study provides evidence of the inability of health information systems to convey the importance of diabetic control to the Asian population. In order that this important information reaches the required recipients, more assertive and perhaps more culturally acceptable methods need to be explored. PMID- 15131684 TI - Laser vs ultrasound biometry--a study of intra- and interobserver variability. AB - BACKGROUND: Accurate biometry is of vital importance in achieving predictable postoperative refraction following cataract surgery. AIM: To evaluate the accuracy and consistency in biometry, achieved by the new generation laser biometric system in comparison with the ultrasound biometric system. METHODS: The study was randomized and prospective. Biometry was performed in 68 eyes of 39 patients by three groups of biometrists (expert, intermediate user, novice). Expert and intermediate users are compared as group A, and expert and novice are compared in group B. Axial length, anterior chamber depth (ACD), and keratometry results are compared by t-test analysis. RESULTS: Axial length measurement variation between expert and non experts was 10 times less using laser than ultrasound (P<0.001). ACD measurement variation was also significantly less when using laser compared to ultrasound (P=0.003). Need for some level of user training is indicated in ACD measurement since group A achieved more consistent readings than group B. Keratometry measurements on the laser system were unreliable due to high range of results. Biometric failure was seen in 12% of eyes undergoing laser and 1% undergoing ultrasound biometry. CONCLUSION: Axial length determination by laser biometry is more accurate and consistent at all levels of biometrist expertise, compared to ultrasound biometry. ACD and keratometry measurements on the laser systems need some degree of user training in order to produce consistent results. PMID- 15131685 TI - Optisol vs Dexsol as storage media for preservation of human corneal epithelium. AB - PURPOSE: To compare the efficacy of two storage media, Optisol GS and Dexsol, in preservation of donor corneal epithelium. METHODS: A total of 12 pairs of corneas not suitable for transplantation, all with intact epithelium, were used in this study, with one cornea of the pair stored in Optisol GS and its other counterpart in Dexsol. At each of three durations of storage--1, 2, and 4 days--four of these paired corneas were prepared for light microscopy and scanning and transmission electron microscopy. Another four pairs of control cornea were prepared in the same way and placed in universal fixative. MAIN OUTCOME MEASURES: Evaluation of the corneas was made by two observers masked as to the identity of the storage medium and length of storage. Loss of epithelial cells was evaluated by light microscopy. The attachment of the epithelium to the basement membrane,cellular integrity, intercellular junctions, and intracellular organelles were evaluated and compared by electron microscopy. RESULTS: The magnitude of epithelial loss correlated with the length of storage time. Control corneas maintained normal epithelium with preservation of all epithelial cell layers. Corneas stored for 1 day had minimal damage of the epithelium. Corneas stored for 2 days had a slight increase in epithelial damage, and corneas stored for up to 4 days showed a marked increase in epithelial damage. There were no significant differences between the two storage media. The basal cell layer was maintained in both the media at all time points, usually in good condition with mild-to-moderate damage in some cases. CONCLUSIONS: Loss of donor epithelium is related mainly to the length of storage and is similar in both Optisol GS and Dexsol. The storage time should be less than 4 days,especially when performing penetrating keratoplasty on patients with ocular surface disorders. PMID- 15131686 TI - Germinoma with synchronous involvement of the pineal gland and the suprasellar region: a treatable cause of visual failure in a young adult. AB - Germinomas constitute 0.1-3.4% of all intracranial tumours and 90% affect patients less than 20 years old. Although they are relatively aggressive tumours, which commonly invade the surrounding tissue and seed within the central nervous system, they are very radiosensitive and carry a favourable prognosis. We describe a 19-year-old man who presented with bilateral visual failure secondary to a germinoma with synchronous involvement of the pineal gland and the suprasellar region. Radiotherapy restored his vision almost back to normal. PMID- 15131687 TI - Frosted branch angiitis: a review. AB - The purpose of this study is to present the first report of a case of primary frosted branch angiitis from the UK and to review the characteristics of this rare disease. Primary frosted branch angiitis causes characteristic florid translucent retinal perivascular sheathing of both arterioles and venules in association with variable uveitis, retinal oedema and visual loss, normally with good recovery. A total of 57 cases have been reported in the world literature. Atypical, typically focal frosted branch angiitis may also occur secondary to other causes of intraocular inflammation, especially cytomegalovirus retinitis. Primary frosted branch angiitis has a characteristic presentation but a variable course, typically affecting children or young adults. The disease is likely to represent a common immune pathway in response to multiple infective agents. The optimal treatment is unclear. PMID- 15131688 TI - The long-term results of nonexpulsive total iridodialysis: an isolated injury after phacoemulsification. PMID- 15131689 TI - Tubulo-interstitial nephritis and uveitis with bilateral optic disc oedema. PMID- 15131690 TI - Severe retinal damage after macular hole surgery with extensive indocyanine green assisted internal limiting membrane peeling. PMID- 15131691 TI - Ocular lesions caused by caterpillar hairs. PMID- 15131692 TI - Oral valganciclovir treatment of varicella zoster virus acute retinal necrosis. PMID- 15131693 TI - Vertical sensory nystagmus associated with intraocular haemorrhages in the shaken baby syndrome. PMID- 15131694 TI - Leukaemic infiltration of the optic nerve as the initial manifestation of leukaemic relapse. PMID- 15131695 TI - Poland anomaly associated with ipsilateral combined hamartoma of retina and retinal pigment epithelium. PMID- 15131696 TI - Topography of the ISW2-nucleosome complex: insights into nucleosome spacing and chromatin remodeling. AB - Linker DNA was found to be critical for the specific docking of ISW2 with nucleosomes as shown by mapping the physical contacts of ISW2 with nucleosomes at base-pair resolution. Hydroxyl radical footprinting revealed that ISW2 not only extensively interacts with the linker DNA, but also approaches the nucleosome from the side perpendicular to the axis of the DNA superhelix and contacts two disparate sites on the nucleosomal DNA from opposite sides of the superhelix. The topography of the ISW2-nucleosome was further delineated by finding which of the ISW2 subunits are proximal to specific sites within the linker and nucleosomal DNA regions by site-directed DNA photoaffinity labeling. Although ISW2 was shown to contact approximately 63 bp of linker DNA, a minimum of 20 bp of linker DNA was required for stable binding of ISW2 to nucleosomes. The remaining approximately 43 bp of flanking linker DNA promoted more efficient binding under competitive binding conditions and was functionally important for enhanced sliding of nucleosomes when ISW2 was significantly limiting. PMID- 15131697 TI - Choline acetyltransferase structure reveals distribution of mutations that cause motor disorders. AB - Choline acetyltransferase (ChAT) synthesizes acetylcholine in neurons and other cell types. Decreases in ChAT activity are associated with a number of disease states, and mutations in ChAT cause congenital neuromuscular disorders. The crystal structure of ChAT reported here shows the enzyme divided into two domains with the active site in a solvent accessible tunnel at the domain interface. A low-resolution view of the complex with one substrate, coenzyme A, defines its binding site and suggests an additional interaction not found in the related carnitine acetyltransferase. Also, the preference for choline over carnitine as an acetyl acceptor is seen to result from both electrostatic and steric blocks to carnitine binding at the active site. While half of the mutations that cause motor disorders are positioned to affect enzyme activity directly, the remaining changes are surprisingly distant from the active site and must exert indirect effects. The structure indicates how ChAT is regulated by phosphorylation and reveals an unusual pattern of basic surface patches that may mediate membrane association or macromolecular interactions. PMID- 15131698 TI - CITRX thioredoxin interacts with the tomato Cf-9 resistance protein and negatively regulates defence. AB - To identify proteins involved in tomato Cf-9 resistance protein function, a yeast two-hybrid screen was undertaken using the cytoplasmic C-terminus of Cf-9 as bait. A thioredoxin-homologous clone, interacting specifically with Cf-9, was identified and called CITRX (Cf-9-interacting thioredoxin). Virus-induced gene silencing (VIGS) of CITRX resulted in an accelerated Cf-9/Avr9-triggered hypersensitive response in both tomato and Nicotiana benthamiana, accompanied by enhanced accumulation of reactive oxygen species, alteration of protein kinase activity and induction of defence-related genes. VIGS of CITRX also conferred increased resistance to the fungal pathogen Cladosporium fulvum in the otherwise susceptible Cf0 tomato. CITRX acts as a negative regulator of the cell death and defence responses induced through Cf-9, but not Cf-2. Recognition of the Cf-9 C terminus by CITRX is necessary and sufficient for this negative regulation. This is the first study that implicates thioredoxin activity in the regulation of plant disease resistance. PMID- 15131700 TI - [Re-defining academic medicine]. PMID- 15131699 TI - Bcl-x(L) sequesters its C-terminal membrane anchor in soluble, cytosolic homodimers. AB - Bcl-x(L) is a potent inhibitor of apoptosis. While Bcl-x(L) can be bound to mitochondria, a substantial fraction, depending on the cell type or tissue, is found in the cytosol of healthy cells. Gel filtration and crosslinking experiments reveal that, unlike monomeric Bax, Bcl-x(L) migrates in a complex of approximately 50 kDa in the cytosol. Co-immunoprecipitation experiments indicate that Bcl-x(L) in the cytosol forms homodimers. The C-terminal hydrophobic tails of two Bcl-x(L) molecules are involved in homodimer formation, and analysis of mutants demonstrates that the C-terminal lysine residue and the G138 residue lining the BH3-binding pocket are required for homodimerization. The flexible loop preceding the C-terminal tail in Bcl-x(L) is longer than that of several monomeric Bcl-2 family members and is a requisite for the homodimer formation. Bad binding to Bcl-x(L) dissociates the homodimers and triggers Bcl-x(L) binding to mitochondrial membranes. The C-terminal tail of Bcl-x(L) is also required to mediate Bcl-x(L)/Bax heterodimer formation. Both mitochondrial import and antiapoptotic activity of different Bcl-x(L) mutants correlate with their ability to form homodimers. PMID- 15131701 TI - [The healthy choices]. PMID- 15131702 TI - [Habilitation of children with cerebral palsy]. PMID- 15131703 TI - [A family-based strategy for diagnosing familial hypercholesterolemia]. AB - BACKGROUND: Patients with familial hypercholesterolaemia have increased risk of developing coronary heart disease. The most cost-effective way of diagnosing patients with familial hypercholesterolaemia is to perform genetic testing of close relatives of already diagnosed patients. MATERIAL AND METHODS: Probands with familial hypercholesterolaemia in whom the underlying mutation in the low density lipoprotein receptor gene has been identified were informed that close relatives should also be tested. RESULTS: Blood samples were taken for molecular genetic testing from 851 first-degree relatives of patients with familial hypercholesterolaemia; 47.8 % tested positively and 52.2 % negatively. Among those with positive tests, only 41.5 % were on lipid-lowering drugs and only 6.1 % had a value for total serum cholesterol < 5 mmol/l. Six months after testing, 81.9 % were on lipid-lowering drugs, at which time a mean reduction in total serum cholesterol of 21.2 % (p < 0.0001) was observed in patients aged 18 and above who were not on treatment at the time of testing. INTERPRETATION: Molecular genetic testing for familial hypercholesterolaemia in a family-based strategy is a cost-effective way of diagnosing patients with this condition that leads to implementation of effective preventive measures. PMID- 15131704 TI - [Complex craniofacial synostoses]. AB - BACKGROUND: Complex craniofacial synostosis is a group of rare genetic disorders characterized by premature closure of the sutures in the craniofacial skeleton and which to varying degrees affects the extremities. MATERIAL AND METHODS: On the basis of relevant literature, we present a review of syndromal craniofacial synostosis. RESULTS: Phenotypically, the complex craniofacial syndromes have many similarities. Synostosis of the sutures of the cranial vault can result in a variety of skull deformations, depending on the sutures involved, the sequence of premature closure, and the time of closure. Synostosis of the sutures in the skull base and facial skeleton leads to shallow orbits, exophthalmus, hypertelorism, midface retrusion, and prognathia. INTERPRETATION: Precise diagnosis of complex craniofacial syndromes may be difficult solely on the basis of a clinical examination. However, several of the most common syndromes are caused by mutations in genes that code for fibroblast growth-factor receptors. Children with a suspected complex craniofacial syndrome should be referred to genetic testing. PMID- 15131705 TI - [Children with cerebral palsy referred to a multidisciplinary habilitation unit]. AB - BACKGROUND: At Akershus University Hospital a new multidisciplinary team was set up to serve neurologically impaired children. We investigated how many children with cerebral palsy were referred to the team and whether referral resulted in an associated diagnosis. MATERIAL AND METHODS: All children with a diagnosis of cerebral palsy referred up until the end of the team's first year of full service were included. Data on birth weight and mobility were retrieved from patient files. Additional diagnoses were registered at inclusion date and two years later and compared to prevalence studies. RESULTS: 182 children had a diagnosis of cerebral palsy. 41 out of 114 children (36%) in the age bracket 6-14 were described as walking without using aids. After a minimum of two years' follow-up, 80 children had no neurological or psychiatric diagnosis (except cerebral palsy). INTERPRETATION: Children with mild motor impairment were underrepresented, and associated impairments, except epilepsy, were diagnosed less often than expected. PMID- 15131706 TI - [Use of stents in abdominal aortic aneurysms]. AB - BACKGROUND: Endovascular treatment of aortic aneurysms has acquired a widespread application. We present the results of endovascular treatment of infrarenal, abdominal aortic aneurysms in our hospital from 1995 through 2002. MATERIAL AND METHOD: Seventy-one stent graft procedures were performed on 69 patients (64 men), mean age 72 years (range 48-96 years). Mean aneurysm diameter was 57 mm (range 35-100 mm). Sixty-nine procedures were elective and two were emergency procedures. All data were registered prospectively. RESULTS: Two procedures failed initially because of technical problems. Both patients underwent a successful procedure later. Immediate conversion to open surgery was done in one case because of a collapse of the graft into the aneurysm sac. There was no 30 day mortality for elective procedures. The conversion rate after a mean observation time of 42 months was 11%. Late complications resulted in 47 re interventions in 29 patients, of which 96% were done in cases treated with Stentor or Vanguard prostheses. CONCLUSION: Endovascular treatment of aortic aneurysms is an alternative to open surgery. Stent graft failure can be serious and difficult to predict. This treatment should not be recommended to patients younger than 70 years and fit for open surgery. PMID- 15131707 TI - [Use and misuse of dietary supplements]. PMID- 15131708 TI - [From a hypothesis to dietary advice--roads, short cuts and dead-end roads]. AB - Few scientific fields receive more public attention than food and nutrition and few have more self-proclaimed experts. Many are confused by the jungle of dietary advice, sometimes widely differing, with the media often giving broad attention to isolated results without assessing them in the context of the total body of knowledge within a field. A new hypothesis may be received with enthusiasm by lay people and professionals alike, but the road to a well-documented conclusion on which to build general advice, is most often long. A finding must be seen in the context of other findings and judged in relation to the whole body of theory within a field before conclusions can be drawn. Work is carried out both nationally and internationally in order to reach a shared understanding of the minimum requirements of documentation before advice can be given. Traditional guidelines for evidence-based medicine can be applied to some but not all aspects of the science of nutrition. This article presents the process leading to official dietary guidelines in Norway. PMID- 15131709 TI - [Health claims about food]. AB - Health claims about food requires harmonised guidelines; the EU has started work on a regulation. The purpose of health claims should be to help consumers select foods which they can include as a natural part of their diet and which benefit their health. Importantly, the use of health claims on some foods should not undermine confidence in normal traditional foods, which is not branded or marketed with such claims. The introduction of health claims will not replace the efforts at providing general information and education aimed at consumers. Food companies have a special interest in including health claims in their marketing. However, there are as yet no studies demonstrating that such claims promote health in the general population or even healthy eating, hence the importance of a thorough evaluation of health claims. PMID- 15131710 TI - [Is decentralisation of specialised healthcare beneficial in social and financial terms?]. AB - BACKGROUND: This article examines the issues of whether decentralisation of specialised healthcare is socially and economically beneficial, who benefit from it, and who carry the costs. MATERIAL AND METHOD: Data for this study was drawn from an out-patient clinic in the town of Alta in northern Norway. A cost analysis was done based on 5026 consultations in an out-patient clinic over a six month period in 2002. RESULTS: We estimated the additional costs of running the out-patient clinic at NOK 4.5 m. The alternative provision of these services implies that the patient would have had to travel to hospital for medical treatment, with costs estimated at NOK 14.1 m. The estimated cost savings over the period were accordingly NOK 9.6 m. Three parties benefit from decentralisation: patients and those who would have had to come to a hospital with them, the National Insurance System, and employers. Finnmark Health Enterprise carried the additional costs of running the out-patient facility. The estimated public-sector cost savings from decentralisation over this six-month period were, at a minimum, NOK 0.8 m. INTERPRETATION: The analysis reveals shortcomings in the system for financing specialised healthcare. As long as National Insurance System covers travelling expenses, the health enterprise does not stand to save money from decentralisation, though it is socially and economically efficient when all factors are taken into account. PMID- 15131711 TI - [Evolution, diet and life style]. PMID- 15131712 TI - [Organization and responsibilities of hospital pharmacies]. PMID- 15131714 TI - [Small lives in a bigger conflict]. PMID- 15131715 TI - [Physicians and expert testimony]. PMID- 15131716 TI - [Norwegian cardiology--some reflections by an epidemiologist]. PMID- 15131717 TI - [Norwegian cardiology in 2004]. PMID- 15131718 TI - [Errors in a textbook on psychiatry]. PMID- 15131720 TI - [Immigrants in an emergency psychiatric department]. PMID- 15131722 TI - [Treatment of urinary incontinence and nocturnal enuresis in primary health care]. PMID- 15131725 TI - Photostability of naturally occurring whitening agents in cosmetic microemulsions. AB - An o/w microemulsion formulated using lecithin and an alkyl glucoside as mild, non-irritant surfactants was proposed as a cosmetic vehicle for arbutin and kojic acid, naturally occurring whitening agents. After assessing the physicochemical stability of the microemulsion in the presence and absence of whitening agents, several perfumed compositions, developed using fragrant molecules of natural or synthetic origin, were introduced, and the olfactory impact of the perfumed microemulsion was evaluated. The photostability to UVB irradiation of both whitening agents was determined in aqueous solutions and in microemulsions, and also in the presence of the perfumed compositions. The stability of arbutin and kojic acid was higher in microemulsions than in aqueous solutions, and only in some cases did the presence of odorous molecules appear to influence it: linalool exerted some protective effect towards kojic acid photodegradation. PMID- 15131726 TI - Normal human epidermal keratinocytes treated with 7-dehydrocholesterol express increased levels of heat shock protein. AB - Human skin, and its isolated cells, respond to insults with a variety of repair and protective mechanisms. One such mechanism is the production of heat shock proteins (HSPs). Heat shock proteins help the other cellular proteins fold correctly into their active three-dimensional structures. Therefore, they can enhance the survival of cells under harsh, denaturing conditions. In order to develop a means of promoting the heat shock response to prepare the skin to withstand insult, we are investigating materials that appear to protect the skin biologically. One such material is vitamin D3 and its precursors. We have observed that keratinocytes treated with 7-dehydrocholesterol (7-DHC), a precursor of vitamin D3, have increased levels of protein and mRNA for heat shock proteins. In addition, we observed that topically applied 7-DHC increases the minimal dose of UVB required to induce erythema. These data suggest that 7-DHC can induce heat shock proteins in skin keratinocytes and that they will be more resistant to UVB insult. PMID- 15131727 TI - Lack of burning and stinging from a novel first-aid formulation applied to experimental wounds. AB - Antiseptic-anesthetic first aid formulations typically produce a burning or stinging sensation when used on wounds. The aim of this study was to assess the lack of sting/burn potential of a prototype first-aid wipe when applied to superficial wounds. This was a one-day, double-blind, randomized study, evaluating the stinging/burning sensation from a prototype first-aid wipe with a cellulose base containing 1.0% pramoxine HCL and 0.13% benzalkonium chloride. The study followed a direct comparison test design of four test formulations. The remaining three articles were: (a) sterile 0.9% sodium chloride (no-sting/no-burn control), (b) 3% hydrogen peroxide (first-aid sting/burn control), and (c) 70% isopropyl alcohol (second sting/burn control). The test was performed on 24 subjects. The tape stripping method was used to create four standardized wounds on the volar forearms, reaching the glistening layer. Each test article was applied to the wound for 15 seconds. The subjects were asked to report the intensity of the stinging/burning sensation during the application. The prototype pramoxine-benzalkonium chloride wipe produced significantly less stinging/burning than both 70% isopropyl alcohol and 3% hydrogen peroxide. Also, the prototype wipe did not produce more stinging and burning in superficial wounds compared to the saline control. PMID- 15131728 TI - Salivary films on hydroxyapatite studied by an in vitro system for investigating the effect of metal ions and by a quartz-crystal microbalance system for monitoring layer-by-layer film formation. AB - The salivary film or the acquired pellicle is a protein film formed initially on the enamel surface of teeth. Such a film plays an important role in enamel protection, but is also an initial substructure for the formation of plaque and the cosmetically undesirable colored stain. The composition and the structure of the film are still essentially unknown because of the difficulty of its isolation for characterization. The purpose of this study was to investigate the effect of some metal cations on the salivary film or the pellicle formation, and also to clarify the mechanism of development. First, using infrared spectroscopy (IR) and X-ray photoelectron spectroscopy (XPS), the in situ-formed film in the mouth was confirmed to contain selectively adsorbed well-known proteins. Then, in vitro studies have demonstrated that Ca2+ ions enhance film formation at the initial stage in virtue of Ca bridging and, interestingly, that Mg2+ ions oppositely inhibit the formation. Furthermore, the quartz-crystal microbalance (QCM), utilized successfully for the first time to study the salivary film, has shown the possibility of an alternate accumulation mechanism by which the surface charges on the film are effectively reversed by the opposite charged proteins. PMID- 15131729 TI - Delivery of vitamin E to the skin by a novel liquid skin cleanser: comparison of topical versus oral supplementation. AB - Topical supplementation represents an attractive approach to mitigate environmentally induced deficiencies of skin vitamin E (alpha-tocopherol). We report here the impact of natural sunlight on stratum corneum (SC) vitamin E and also compare the effectiveness of dietary supplementation to topical application as a way to increase vitamin E in the superficial layers of the SC. The effects of natural sunlight, 30 minutes of midday sunlight, were measured on two separate occasions. Vitamin E in the surface layers of the SC was measured by HPLC after ethanol extraction. Under these relevant conditions, vitamin E in the superficial SC was reduced in a dose-dependent manner by 50-65%. In a followup study, panelists entered into a randomized, double-blind, vehicle-controlled study. In this study, one group washed their skin once daily for one minute with a commercially available body wash containing 0.15% vitamin E and 0.10% vitamin E acetate, while the second group used a body wash without vitamin E but also supplemented their diet with 400 IU alpha-tocopherol (18 x RDI). Not surprisingly, only dietary supplementation increased serum vitamin E (approximately twofold). Although both treatment modalities increased SC vitamin E, topical delivery was significantly more effective (53-fold vs baseline) than dietary delivery (eightfold vs baseline). Moreover, only topical delivery increased SC vitamin E acetate (19-fold vs baseline). The results reported here indicate that vitamin E in the superficial layers of the SC is depleted readily by even a brief exposure to sunlight and that use of a vitamin E body wash can substantially increase the vitamin E in this superficial layer more effectively than dietary supplementation. PMID- 15131730 TI - Clinical effects of cosmetic vehicles on skin. PMID- 15131732 TI - [Thrombolytic treatment in acute brain infarction]. PMID- 15131733 TI - [Evaluation of cerebral blood perfusion in patients undergoing carotid endarterectomy using densitometric analysis of conventional cranial CT]. AB - OBJECTIVE: To evaluate the efficacy of densitometric analysis of cranial CT imaging in the measurement of areas with low cerebral blood flow and lack of hemodynamic reserve in patients undergoing carotid endarterectomy. PATIENTS AND METHODS: 40 consecutive patients undergoing carotid endarterectomy have been included in the study. All of them had preoperative cranial CT, pre and postoperative basal and acetazolamide SPECT. Cranial CT imaging after digitalization and computer processing were obtained with 4 densitometric patterns: 1). normal cerebral blood perfusion; 2). ischemic or low brain blood perfusion (patron I); 3). parenchyma without cerebral hemodynamic reserve (patron nR), and 4). brain infarction. RESULTS: 32 out of 40 (80 %) patients had abnormal densitometric patterns: 19 nR patterns (47.5 %), 11 I pattern (27.5 %) and 2 established brain infarction. The correlation between densitometric cranial TC imaging and SPECT was 92% when analyzing areas with hypoperfusion (S: 96.5 %; E: 82 %). Cranial CT detected 12/13 of patients who lacked cerebral hemodynamic reserve in the SPECT analysis (S: 74 %; E: 92 %; Vpp: 95 %). CONCLUSIONS: There is a good correlation between analysis of densitometric cranial TC imaging and SPECT in determining low cerebral blood flow areas and lack of cerebral hemodynamic reserve. This method could help to improve carotid surgery indications in patients with carotid stenosis. PMID- 15131734 TI - [Verbal fluency: preliminary normative data in a Spanish sample of young adults (20-49 years of age)]. AB - INTRODUCTION: Verbal fluency tests are widely used to study, among other things, executive functions. The aim of this study is to establish pilot normative data of these tests (phonological and semantic fluency) for a healthy young adult Spanish population as well as to analyze the influence of age and years of formal education on test performance. SUBJECTS: A total of 146 healthy young subjects (43.2 % male and 56.8 % female) were administered the tests. Age varied between 20 and 49 (mean age: 34.21 years) and years of formal education ranged from 8 to 20 (mean: 14.51 years). METHOD: Phonological fluency tests (words starting with the phoneme F, A and S; words not containing A, E and S) and semantic fluency tests (fruits and vegetables, animals, tools) were administered. Performance within one minute was registered as total as well as by quartiles. Statistical analysis was based on the descriptive analysis of the sample. Influence of the variables age and education on each category was studied by simple regression analysis. RESULTS: The variable formal years of education significantly influenced the scores of all independent variables. Scores were adjusted accordingly. Age was significant for the categories <> and <>. Correlation criteria for the variables that test performance were obtained. CONCLUSIONS: Preliminary normative data obtained in the present study will be useful for an adequate neuropsychological assessment of young adult patients with regards to verbal fluency measures. Limitations of some of the categories are discussed. PMID- 15131735 TI - [Functional recovery and return to work in the young patient after a stroke]. AB - INTRODUCTION: There are few works that study the functional recovery of young patients after suffering cerebrovascular disease (CVD) and even fewer works that study work reincorporation. Our aim is to evaluate these two aspects in the middle term in the young adults who have suffered a stroke and also to establish the factors that can influence a better prognosis. PATIENTS AND METHODS: Our work is a cross-sectional descriptive study of 111 patients aged between 15 and 55 years who have been discharged from our Hospital with a CVD diagnosis during the years 1999-2000. We have collected the following data from the clinical records: age, gender, type, location and reason for the CVD, vascular risk factors, complications in the process, with or without rehabilitation treatment and treatment description. By means of telephone interview, we obtained functional recovery with the Barthel Index (BI) and Modified Rankin Scale (MRS) as well as study level, marital status and pre and poststroke labor situation, loss of friends, loss of recreational activities and finally, sequels after the discharge. RESULTS: Functional recovery for daily activities (IB >or= 90) was good in 78 % of the patients, this being 72.1 % if we assess their global handicap (MRS >, especially focused on the autosomal recessive dystrophies, taking the study of dystrophinopathies as an example. PMID- 15131737 TI - [Neuropsychology of Korsakoff's syndrome]. AB - The main manifestation of Korsakoff's syndrome is a disproportionate impairment in memory in comparison to other cognitive functions. Most prominent is anterograde amnesia for both verbal and nonverbal information. The marked sensitivity to interference is the main characteristic of memory impairment of these patients. Retrograde amnesia is also a typical feature of this disorder. Memory for autobiographical information as well as knowledge of public events and facts are affected and they have a temporal gradient. Confabulations may also occur but they are not specific to Korsakoff's syndrome. Together with the disproportionate deficits in memory that define this disorder, different grades of involvement of other cognitive disorders are observed. The aim of this paper is to review the most important neuropsychological features of Korsakoff's syndrome and to discuss the hypotheses on it. PMID- 15131738 TI - [Atherothrombotic carotid disease: towards a consensus on its prevention]. AB - This consensus document was one of the objectives of the I Meeting of the Spanish Society of Angiology and Vascular Surgery, the Spanish Interventional Neuroradiology Group of the Spanish Society of Neuroradiology, and the Cerebrovascular Disease Study Group of the Spanish Society of Neurology, which was held in October 2002 in Cordoba. Atherosclerosis is a chronic vascular disease of true epidemic proportions. It is the first cause of death in developed countries and responsible for one quarter of documented deaths worldwide. Atherothrombotic ischemic stroke, transient ischemic attack and atherothrombotic origin symptomatic or asymptomatic peripheral arterial disease are all associated with a high risk of vascular death, myocardial infarction and recurrent stroke. In this context, vascular disease represents a serious public health problem, particularly if we take into account current forecasts on population ageing. The prevention of atherosclerosis - and its consequences, if its clinical manifestations are already apparent - is therefore a priority. This multidisciplinary consensus document draws on the different medical specialties dealing with these patients and combines efforts to obtain the greatest possible benefit as regards this disease's management. The consensus document makes a global analysis of atherosclerosis prevention, basically in terms of therapeutic objectives, lifestyle change measures, high bloodpressure management, dyslipidemia, other vascular risk factors and platelet antiaggregation. Emphasis is placed on the frequent coexistence of cerebrovascular and peripheral arterial involvement, and the methods of detecting silent involvement. Lastly, consensus is reached on the diagnostic methods and specific management of atherothrombotic carotid disease, including the benefits and risks of and indications for carotid endarterectomy, and the current role of carotid angioplasty. PMID- 15131739 TI - [Multiple sclerosis and acute weakness of the bulbar and respiratory muscles]. PMID- 15131740 TI - [Ring chromosome 20: a distinctive syndrome identifiable by electroclinical diagnosis]. AB - The ring chromosome 20 syndrome is characterized by treatment resistant non convulsive status epilepticus, and slow waves intercalated by spikes/spike waves predominantly in the front-temporal regions. Here, we describe the case of an 18 year old patient, whose seizures began at the age of 10, these being resistant to treatment. Neurologic examination and cranial MRI were normal. Interictal EEG showed normal background activity with burst of 2-20 seconds with bilateral spike wave. Ictal EEG showed continuous paroxysmal activity with generalized spike waves discharges and slow delta waves, coinciding with nonconvulsive status epilepticus. After 1 mg of intravenous clonazepam, both clinical semiology and EEG abnormalities disappeared. A cytogenetic study showed ring chromosome 20 in 35 % of metaphases. The epilepsy associated with ring chromosome 20 constitutes a syndrome with its distinctive electroclinical characteristics. PMID- 15131741 TI - [Multifocal motor neuropathy with conduction blocks and prurigo nodularis. A paraneoplastic syndrome in a patient with non-Hodgkin B-cell lymphoma?]. AB - Multifocal motor neuropathy with conduction blocks (MMNCB) is a peripheral demyelinating neuropathy. The etiology of this disease is unknown, but an autoimmune origin is postulated. Prurigo nodularis (PN), a chronic dermatosis also having an unknown etiology and many peripheral neuropathies of different nature are associated to hematological tumors. We have found no cases in the literature in which MMNCB was presented as a paraneoplastic syndrome of a non Hodgkin B-cell type lymphoma (NHL-B). We present the case of a 67 year old man who simultaneously developed PN and MMNCB in upper limbs and who was diagnosed of a NHL-B 19 months later. We raise the hypothesis that both prurigo and neuropathy are a paraneoplastic syndrome for lymphoma with a possible common autoimmune pathogenic mechanism. PMID- 15131742 TI - [Asterixis in focal brain lesions]. AB - Asterixis is a motor control disorder characterized by the presence of abnormal movements of the lower limbs in the vertical plane during posture maintenance. Asterixis is usually bilateral and associated with toxic-metabolic metabolic encephalopathies. Unilateral asterixis is less frequent and it normally indicates focal brain damage. We report the cases of four patients (two males/two females), aged 57 to 83 years, suffering from uni or bilateral asterixis associated with focal brain damage. All patients underwent CT brain scan and a neurophysiological study (parietal EMG and/or PES). In addition, any toxic-metabolic cause that could be produced by this clinical phenomenon was ruled out with the appropriate testing. Unilateral asterixis is a clinical symptom that may indicate the presence of focal brain damage. Often, it is ignored or overlooked during routine neurological examinations. On the other hand, the presence of a bilateral asterixis is not always indicative of a toxic-metabolic encephalopathy.Rarely, such as in one of the cases herein presented, bilateral asterixis can also appear associated with structural brain lesions. Although asterixis diagnosis is fundamentally clinical, the neurophysiological study contributes to verify the diagnosis. PMID- 15131743 TI - [Unstable angina in relationship to intake of zolmitriptan]. PMID- 15131744 TI - [Based on Foix-Chavany-Marie syndrome]. PMID- 15131745 TI - [Early symptoms and causes of sudden death related to sports activities]. AB - BACKGROUND: This study provides an overview on presymptoms and causes of sport related death and suggests preventive measures. METHODS: 29 436 autopsies of the Centre of Legal Medicine at the University of Frankfurt/Main from 1972 to 2001 were analysed and compared with a control group. RESULTS: 96 men (95 %, average age 53.3 years) and 5 women (5 %, average age 36.4 years) were involved in 101 fatalities connected with sports (0.34 % of all autopsies). 78 athletes (77.2 %) died during, 20 (19.8 %) immediately after sport activities. Immediately before death 18 of 50 persons concerned were free of symptoms, twelve complained of nausea and vomiting, seven of vertigo, five of weakness, four of epigastric pain and dyspnoea, three exhibited angina pectoris symptoms. The day before death 7 of 25 athletes had symptoms like angina pectoris, nausea, backache and palpitation. 84 fatalities (83.2 %) were due to coronary heart disease. Seven were due to myocarditis (6.9 %), in four cases (26.7 %) among athletes younger than 36. Among children and adolescents (n = 4) myocarditis, cardiomyopathy and in two cases a valvular defect of the heart were the cause of death. In comparison to controls death in elder athletes due to coronary heart disease and myocardial infarction was high significantly more common and athletes died significantly younger. CONCLUSIONS: Further prospective investigations are needed to provide strategies for prevention. PMID- 15131746 TI - [Sustained weight reduction after cessation of obesity treatment with Sibutramine]. AB - BACKGROUND AND AIM OF THE STUDY: Aim of the study was to examine the course of body weight and cardiovascular risk parameters under the everyday life conditions in general practice after obese patients had terminated their sibutramine treatment. PATIENTS AND METHODS: Patients were administered sibutramine (10 mg oder 15 mg, once daily) for at least six months could be included in the post marketing surveillance (PMS) study. Control visits to document body weight, laboratory parameters and blood pressure took place after 12 weeks and after 24 weeks. RESULTS: In all, 374 patients (77 % women, 20 % men) with an average age of 45.8 +/- 11.8 years terminated the previous sibutramine therapy at the latest with inclusion into the study. On average, they had administered sibutramine for six months. At the end of the 24-week observation period the majority of the patients stated that they had continuously adhered to changed life habits during the last six months. On average, the body weight of the patients did not increase (mean weight reduction of 1 kg between start and end of the observation period). The stable course of the body weight was mirrored by the values of the laboratory parameters: values for total cholesterol, HDL cholesterol, LDL cholesterol, triglycerides, serum glucose (fasting), HbA (1c), urea, uric acid and liver parameters remained unchanged compared to the values on entry into the study. Throughout the observation period, blood pressure values remained stable as well. CONCLUSION: Values for body weight, vital signs (blood pressure, heart rate), serum glucose and serum lipids remain constant even after termination of the sibutramine therapy. Possibly, the positive effect on body weight reduction by the treatment with sibutramine support the adherence to a changed life style. Obviously, this action continues after termination of the drug treatment. PMID- 15131747 TI - [Iatrogenic perforation of the esophagus during transesophageal echocardiography. A rare cause of severe dysphagia]. AB - HISTORY: A 65-year-old patient underwent transesophageal echocardiography which caused a perforation of the upper esophagus. Three months after esophagostomy and gastrostomy the reconstruction was accomplished by a colon interposition graft. The patient postoperatively developed an ischemic necrosis of the graft, followed by a cervical fistula. Food intake and swallowing became impossible. DIAGNOSIS: X ray examinations revealed the cervical fistula and a stenotic colon graft. TREATMENT AND COURSE: The retrosternal colon graft was replaced by a gastric interposition graft, which was anastomosed with the cervical esophagus. The postoperative follow-up was normal at first. Increasing retention of secretion in the remaining esophagus however caused dilatation and a cervival fistula again, as well as a pleural empyema. After transthoracic resection of the esophagus the patient was finally free of symtoms, and gained weight on unrestricted food intake. CONCLUSION: Transesophageal echocardiography is a common diagnostic procedure with a low complication rate. Even though serious complications may occur in rare cases, the patient must be informed about the risk. The perforation of the esophagus is an emergency situation that requires surgical treatment immediately. Primary reconstruction and preservation of the esophagus is the recommended strategy. PMID- 15131748 TI - [Assessment of myocardial viability following heart infarct--case report]. PMID- 15131749 TI - [Assessment of myocardial viability following heart infarct]. PMID- 15131752 TI - [Gastrointestinal side effects of drugs]. PMID- 15131753 TI - [Short guideline for the management of dyspepsia]. PMID- 15131754 TI - Linkage analysis of a complex disease through use of admixed populations. AB - Linkage disequilibrium arising from the recent admixture of genetically distinct populations can be potentially useful in mapping genes for complex diseases. McKeigue has proposed a method that conditions on parental admixture to detect linkage. We show that this method tests for linkage only under specific assumptions, such as equal admixture in the parental generation and admixture that occurs in a single generation. In practice, these assumptions are unlikely to hold for natural populations, resulting in an inflation of the type I error rate when testing for linkage by this method. In this article, we generalize McKeigue's approach of testing for linkage to allow two different admixture models: (1) intermixture admixture and (2) continuous gene flow. We calculate the sample size required for a genomewide search by this method under different disease models: multiplicative, additive, recessive, and dominant. Our results show that the sample size required to obtain 90% power to detect a putative mutant allele at a genomewide significance level of 5% can usually be achieved in practice if informative markers are available at a density of 2 cM. PMID- 15131755 TI - Increased leucine turnover in women during the third trimester of uncomplicated pregnancy. AB - Plasma amino acid concentrations decrease in pregnancy despite increased requirements for both maternal and fetal protein accretion. There are few published data on changes in amino acid turnover in pregnancy. The aim of this study is to test the hypotheses that (1) whole body nonoxidative leucine disposal (NOLD) is higher and (2) whole body oxidative leucine disposal (OLD) is lower in the third trimester of pregnancy than in the nonpregnant state. After an overnight fast 8 pregnant women between 33 and 35 weeks gestation had a primed infusion of labeled [1-(13)C] leucine and a prime dose of NaH(13)CO(3). Carbon dioxide production was measured using indirect calorimetry. Gas chromatography mass spectrometry was used to determine (13)CO(2) enrichment of expired air and oxidative and nonoxidative leucine turnover by measuring (13)C keto-isocaproate plasma enrichment, which reflects intracellular leucine enrichment. Women acted as their own controls after the puerperium. Whole body leucine turnover expressed per unit body weight was increased in pregnancy (median [interquartile range or IQR]: pregnant = 103.1 [14.9] v nonpregnant = 90.1 [10.9] micromol/kg/h). The mean (+/-SD) of the differences was 11.4 +/- 5.6 micromol/kg/h, P =.0006. NOLD was increased in pregnancy (pregnant = 86.8 [10.1] v nonpregnant = 73.3 [9.5] micromol/kg/h). The mean (+/-SD) of the differences was 10.6 +/- 5.4 micromol/kg/h, P =.0008. OLD was not significantly altered in pregnancy (pregnant = 17.3 [4.5] v nonpregnant = 15.91 [2.4] micromol/kg/h). The mean (+/-SD) of the differences was 0.84 +/- 1.94 micromol/kg/h, P =.26. In conclusion, women have significantly higher NOLD in the third trimester of pregnancy than when not pregnant but there are no significant changes in OLD. PMID- 15131756 TI - The relationship between glycemic control and plasma vascular endothelial growth factor and endothelin-1 concentration in diabetic patients. AB - Understanding the causes of diabetic vascular complications has become an increasingly important issue because of the rapidly rising prevalence of diabetes. Recently discovered vasoconstrictors and angiogenesis regulators, such as endothelin (ET) and vascular endothelial growth factor (VEGF), have been intensely studied for possible pathogenic roles in diabetic vascular complications. The present study was undertaken to clarify the effect of glycemic control on serum VEGF and plasma ET-1 concentrations in diabetic patients, and to identify other factors that may cause fluctuations of these substances. Plasma VEGF and ET-1 concentrations of 45 hospitalized diabetic patients and 54 control subjects were measured by enzyme immunoassay (EIA) and radioimmunoassay (RIA), respectively. Plasma VEGF was elevated in poorly controlled diabetic patients compared with healthy subjects and plasma VEGF concentrations declined after hospitalized treatment with either insulin or oral hypoglycemic agents in combination with diet. There was a significant correlation between plasma VEGF concentration and both fasting plasma glucose (FPG) and hemoglobin A(1c) (HbA(1c)). Plasma ET-1 in poorly controlled diabetic patients was higher than in healthy controls, but improved glycemic control did not affect plasma ET-1 concentrations. Thus, poor glycemic control causes increased levels of plasma VEGF, which may result in hypertension and vascular complications in diabetes. Short-term treatment resulting in good glycemic control can improve levels of VEGF and may provide beneficial effects on diabetic vascular complications. PMID- 15131757 TI - Hydrophilic 7 beta-hydroxy bile acids, lovastatin, and cholestyramine are ineffective in the treatment of cerebrotendinous xanthomatosis. AB - We compared the effect of treatments with hydrophilic bile acids (ursodeoxycholic and ursocholic acids), cholestyramine, and lovastatin versus chenodeoxycholic acid in 4 patients with cerebrotendinous xanthomatosis (CTX). Bile acids and bile alcohols in plasma, bile, and urine before and after treatment were quantitated by gas-liquid chromatography. Untreated, all patients showed abnormal biliary bile acid composition: cholic acid (72.7%) and chenodeoxycholic acid (6.2%), and polyhydroxylated C(27)-bile alcohols (10.0%), and elevated plasma cholestanol levels. Treatment with hydrophobic chenodeoxycholic acid inhibited abnormal bile acid synthesis (virtual disappearance of C(27)-bile alcohols from plasma, bile, and urine and marked reduction of plasma cholestanol levels). Hydrophilic ursodeoxycholic and ursocholic acids did not inhibit abnormal bile acid synthesis, while cholestyramine increased abnormal bile acid synthesis (continued increased formation of polyhydroxylated C(27)-bile alcohols and further elevation of plasma cholestanol levels). Lovastatin did not affect abnormal bile acid synthesis or reduce plasma cholestanol levels. The results demonstrate that impaired side-chain oxidation in bile acid synthesis due to mutations of Cyp27 results in increased formation of polyhydroxylated C(27)-bile alcohols and cholestanol in CTX. Hydrophobic chenodeoxycholic acid, but not cholestyramine, lovastatin, or hydrophilic 7beta-hydroxy acids, inhibited the abnormal synthetic pathway. The role of chenodeoxycholic acid in downregulating abnormal bile acid synthesis in CTX is emphasized. PMID- 15131758 TI - Effects of the selective estrogen receptor modulator LY117018 on growth hormone secretion: In vitro studies. AB - Sex steroids play an important role in modulating pulsatile growth hormone (GH) release, acting at both hypothalamic and pituitary level in both humans and experimental animals. Selective estrogen receptor modulators (SERMs) act as either estrogen receptor agonists or antagonists in a tissue-selective manner. In postmenopausal women, serum GH levels correlate positively with endogenous estradiol levels and insulin-like grwoth factor-I (IGF-I) is positively related to bone mineral density (BMD) at the spine and hip. The aim of the present study was to evaluate, for the first time, the direct effect of LY117018, an analog of raloxifene, on GH secretion from both human and rodent pituitary cells in vitro. Our results demonstrated that pharmacological concentrations of the raloxifene analog LY117018 can stimulate GH secretion through a direct action on the pituitary. LY117018 also showed an estrogen-like activity, inducing the proliferation of rat pituitary GH-secreting adenomatous cells (GH1). PMID- 15131759 TI - Erythrocyte-derived measures of membrane lipid composition in healthy men: associations with arachidonic acid at low to moderate but not high insulin sensitivity. AB - The lipid composition of erythrocyte membranes was explored as a surrogate for that of skeletal muscle in investigations into the influence of membrane fatty acid composition on insulin sensitivity. In a preliminary study (study 1), erythrocyte and monocyte/platelet membrane fatty acid percentages were compared with those of muscle membrane in 10 otherwise healthy men undergoing orthopedic surgery. In a further study (study 2), relationships between erythrocyte membrane fatty acid concentrations and insulin sensitivity, S(I), measured using the intravenous glucose tolerance test (IVGTT), were evaluated in 30 asymptomatic men. In study 1, significant positive correlations were found between muscle and erythrocyte membrane fatty acid percentages for 16:0 saturated fatty acid (r = 0.92, P <.001), and for the 18:2n-6, 20:4n-6, 20:5n-3, and 22:5n-3 polyunsaturated fatty acids (PUFAs) (r = 0.67 to 0.83, P <.05 to.01). There were fewer and weaker associations between muscle and monocyte/platelet membrane fatty acid compositions. In study 2, highly insulin-sensitive individuals (n = 8) had significantly lower erythrocyte membrane fatty acid concentrations than those with low/normal S(I). Among those with low/normal S(I) (n = 22), S(I) correlated positively with erythrocyte membrane arachidonic acid concentration (r = 0.57, P <.01) and with total PUFAs (r = 0.46, P <.05). Indices of delta 6 and delta 5 desaturase activities were significantly higher and lower, respectively, in high compared with low/normal S(I) individuals. For a range of fatty acids, erythrocyte membrane fatty acid composition shows close associations with that of muscle membranes. Measurements in erythrocyte membranes support a role for membrane arachidonic acid content in the modulation of insulin sensitivity, specifically at low/normal insulin sensitivities. PMID- 15131760 TI - Acute effects of nicotine on serum glucose insulin growth hormone and cortisol in healthy smokers. AB - Cigarette smoking impairs glucose tolerance and alters serum levels of hormones involved in glucose metabolism, but the role of nicotine in such hormonal alterations is not well understood. In order to isolate the effects of transdermal nicotine on serum glucose, insulin, growth hormone, and cortisol in smokers, we conducted a randomized double-blind placebo-controlled cross-over study involving 34 healthy volunteer smokers between 18 and 55 years of age. Administration of a 14-mg transdermal nicotine patch resulted in nonsignificantly lowered fasting quantitative insulin-sensitivity index (P =.11) and a nonsignificant 9.3-mg/dL mean increase in serum glucose levels during a 75-g oral glucose tolerance test (OGTT) at time 60 minutes (P =.12). There were no substantial differences between groups in the areas under the curve (AUCs) for glucose (P =.33) or insulin (P =.79) during the OGTT. Levels of insulin and cortisol also were not significantly altered by nicotine. A secondary finding observed in the overall study group (primarily in females) was that nicotine caused a 29% median decrease in serum growth hormone (P =.02). We conclude that nicotine patches may lead to mild hyperglycemia and lowered insulin sensitivity. Further research is needed to determine the clinical implications of the unexpected finding that nicotine decreased growth hormone levels in female smokers. PMID- 15131761 TI - Postprandial plasma fructose level is associated with retinopathy in patients with type 2 diabetes. AB - The aim of the present study was to investigate the association of fructose on microangiopathy in patients with diabetes. Postprandial plasma fructose concentrations and postprandial plasma glucose concentrations were simultaneously measured 3 times within a 24-hour period (2 hours after each meal) in 38 patients with type 2 diabetes that had been admitted to the hospital. The mean postprandial plasma fructose concentrations (MPPF) and the mean postprandial plasma glucose concentrations (MPPG) were calculated. Fructose was measured by gas chromatography-mass spectrometry (GCMS). Based solely on MPPF, we were able to divide the patients into three groups: the high MPPF (31.9 +/- 6.5 micromol/L) group (n = 12), the middle MPPF (21.2 +/- 1.8 micromol/L) group (n = 13), and the low MPPF (15.2 +/- 2.4 micromol/L) group (n = 13). Prevalence and degree of retinopathy and nephropathy were then evaluated in the 3 different groups. A significant correlation was observed in the prevalence of proliferative diabetic retinopathy (PDR) among the 3 MPPF groups (P =.024). The prevalence of PDR was higher in the high MPPF group (75.0%) than in the middle and low MPPF groups (23.1% and 38.5%, respectively). Although not significantly different statistically, the prevalence of all degrees of retinopathy showed a tendency to be higher in the high MPPF group (83.3%) than in the middle and low MPPF groups (46.2% and 46.2%, respectively) (P =.081). Nephropathy prevalence also showed a tendency to be higher in the high MPPF group (66.7%) than in the middle and low MPPF groups (38.5% and 30.8%, respectively), although the differences were not significant. The prevalence of clinical albuminuria was not significantly different among the 3 groups, but there was a tendency for it to be higher in the low MPPF group (30.8%) than in the high and middle MPPF groups (16.7% and 0%, respectively). No significant differences in glycemic indicators and mean duration of diabetes were observed among the 3 groups. The increased prevalence of retinopathy in the high MPPF group suggests that fructose is associated with retinopathy in patients with type 2 diabetes. PMID- 15131762 TI - Serum adiponectin is associated with high-density lipoprotein cholesterol, triglycerides, and low-density lipoprotein particle size in young healthy men. AB - The chromosomal localization of adiponectin has been found to be mapped to human chromosome 1q21.4-1q23, a region that was identified as a susceptibility locus for familial combined hyperlipidemia and polygenic type 2 diabetes. As these 2 disorders are associated with low high-density lipoprotein (HDL)-cholesterol, high triglycerides, and insulin resistance (IR), we examined the relation of serum adiponectin concentrations to serum lipid and lipoprotein profiles as well as IR in young healthy men. Serum adiponectin levels were positively associated with HDL-cholesterol, apolipoprotein (apo) A1, and low-density lipoprotein (LDL) particle size, and negatively associated with triglycerides and apo B. Negative associations were also found between adiponectin and body mass index (BMI), percent body fat, and IR,as determined by homeostasis model assessment (HOMA). However, after adjustment for BMI, no significant associations were found between adiponectin and LDL particle size and apo B. In a multiple regression analysis including all variables that showed significant univariate associations with adiponectin, associations of adiponectin with HDL-cholesterol (beta = 0.079, P =.0009), percent body fat (beta = -0.165, P =.002), and serum leptin (beta = 0.291, P =.01) were statistically significant. HDL-cholesterol (beta = 0.077, P =.001), percent body fat (beta = -0.078, P =.03), and LDL size (beta = 0.092, P =.03) emerged as significant and independent determinants of adiponectin after HOMA IR, fasting glucose, triglycerides, and systolic blood pressure (BP) were taken into account. Together, these variables explained 19% of adiponectin variability in the 2 models. HOMA IR did not emerge as a determinant of adiponectin in both models. These findings suggest that in young healthy men hypoadiponectinemia is more closely related to adiposity and dyslipidemia than IR. PMID- 15131763 TI - Effects of betaine in a murine model of mild cystathionine-beta-synthase deficiency. AB - Cystathionine-beta-synthase (CBS) is required for transsulfuration of homocysteine, an amino acid implicated in vascular disease. We studied homocysteine metabolism in mice with mild hyperhomocysteinemia due to a heterozygous disruption of the Cbs gene. Mice were fed diets supplemented with betaine or dimethylsulfonioacetate (DMSA); betaine and DMSA provide methyl groups for an alternate pathway of homocysteine metabolism, remethylation by betaine:homocysteine methyltransferase (BHMT). On control diets, heterozygous mice had 50% higher plasma homocysteine than did wild-type mice. Betaine and DMSA had similar effects in both genotype groups: liver betaine increased dramatically, while plasma homocysteine decreased by 40% to 50%. With increasing betaine supplementation, homocysteine decreased by 75%. Plasma homocysteine and BHMT activity both showed a strong negative correlation with liver betaine. Homocysteinemia in mice is sensitive to a disruption of Cbs and to methyl donor intake. Because betaine leads to a greater flux through BHMT and lowers homocysteine, betaine supplementation may be beneficial in mild hyperhomocysteinemia. PMID- 15131764 TI - Different responsiveness in body weight and hepatic 11beta-hydroxysteroid dehydrogenase (11beta-HSD) type 1 mrna to 11beta-HSD inhibition by glycyrrhetinic acid treatment in obese and lean zucker rats. AB - Tissue-specific dysregulation of 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) activity in obese humans and animals may be associated with obesity and the metabolic syndrome. We investigated the effect of inhibition of 11beta HSD with glycyrrhetinic acid (GE), an effective 11beta-HSD inhibitor, on body weight regulation in obese Zucker rats, which have a defect in the leptin receptor gene. GE (280 mg/kg/d) was administered in drinking water to 8-week-old male Zucker rats for 14 weeks. GE had no effect on food intake or weight gain, and did not affect hepatic 11beta-HSD1 and renal 11beta-HSD2 mRNA levels in obese rats. In contrast, average daily food intake and body weight on week 14 were significantly reduced by GE in lean rats (both P <.0001). Hepatic 11beta-HSD1 and renal 11beta-HSD2 mRNA levels were also significantly decreased by GE in lean rats (both P <.05). GE had no significant effect on plasma corticosterone levels in obese rats but lowered them in lean rats (P <.05). Plasma leptin levels declined in both GE-treated obese and lean rats (both P <.01). In conclusion, long-term GE treatment decreased weight gain in lean Zucker rats but not in obese Zucker rats. These findings suggest that the differing responses of 11beta-HSD1 to GE in obese and lean Zucker rats are closely associated with the different weight-gain responses. Furthermore, the weight-lowering effect of GE may require intact leptin receptors. PMID- 15131765 TI - Fenofibrate prevents obesity and hypertriglyceridemia in low-density lipoprotein receptor-null mice. AB - Our previous study demonstrated that fenofibrate improves both lipid metabolism and obesity, in part through hepatic peroxisome proliferator-activated receptor alpha (PPARalpha) activation, in female ovariectomized, but not in sham-operated, low-density lipoprotein receptor-null (LDLR-null) mice. The aim of this study was to determine whether fenofibrate prevents obesity and hypertriglyceridemia in male LDLR-null mice. Mice fed a high-fat diet for 8 weeks exhibited increases in body and white adipose tissue (WAT) weights and developed severe hypertriglyceridemia compared with mice fed a low-fat control diet. However, these effects were effectively prevented by fenofibrate. Mice given a fenofibrate supplemented high-fat diet showed significantly reduced body weight, WAT weight, and serum triglycerides versus high-fat diet-fed animals. Triton WR1339 study showed that fenofibrate-induced reduction in circulating triglycerides was due to the decreased secretion of triglycerides from the liver. Moreover, the administration of fenofibrate not only resulted in liver hypertrophy and reduction in hepatic lipid accumulation, but also regulated the transcriptional expression of PPARalpha target genes, such as hepatic acyl-coenzyme A (CoA) oxidase and apolipoprotein C-III (apoC-III). Therefore, our results suggest that alterations in hepatic PPARalpha action by fenofibrate seem to suppress diet induced obesity and severe hypertriglyceridemia caused by LDLR deficiency in male mice. PMID- 15131766 TI - Serum leptin, body fat, and nutritional markers during the six months post-kidney transplantation. AB - Leptin is a 16-kd protein that is thought to be a regulator of food intake and body weight. Many previous studies have reported elevated serum leptin levels in renal failure. In this study, we investigated the outcome of serum leptin and its relationship to body fat (BF), dietary intake, nutritional, and inflammatory markers after kidney transplantation (KTx). A total of 41 kidney transplant recipients were followed-up prospectively during 6 months posttransplantation. Serum leptin, albumin, transferrin, and C-reactive protein (CRP) were measured at KTx, 15 days, 3, and 6 months later. Dietary intake and BF were determined at KTx, 3, and 6 months later. A decrease in serum leptin was observed early at day 15 after KTx; this decrease was significant only in patients with BF >/= 30% of body weight. The decrease was maintained at 3 and 6 months after KTx. In multivariate analysis, an independent impact of higher percentage BF at KTx on the decrease of serum leptin was observed. Serum leptin correlated positively with BF. Conversely, no correlation was found between changes of serum leptin and changes of dietary intake. Leptin correlated positively with CRP at KTx, but not after normalization of renal function. Changes of serum leptin levels were not correlated with those of serum albumin levels. In summary, hyperleptinemia at KTx is manifest in patients with a high percentage of BF. An early and maintained correction follows KTx. Serum leptin levels did not appear to affect alimentary intake at and after KTx. PMID- 15131767 TI - Self-monitoring of plasma triglyceride levels to evaluate postprandial response to different nutrients. AB - Self-monitoring of plasma triglycerides (TG) may be a very useful tool to monitor, on a daily basis, the TG responses to different nutrients, particularly carbohydrates (CHO) and fat, whose influence on postprandial TG levels is not very well known. Therefore, the aim of the present study was to evaluate the TG response of hypertriglyceridemic patients to a similar amount of calories deriving from different sources of CHO and fat. Thirty-nine hypertriglyceridemic patients were randomly assigned to 1 of 2 experimental groups. In 1 group (the fat group), patients were given a standard meal plus a fat supplement of 300 kcal derived from different types of fat (butter, sunflower margarine, olive oil) for dinner, once a week for 3 weeks. In the other group (the CHO group), patients consumed the same standard meal plus a supplement of 300 kcal derived from different types of CHO (bread, coke, fruit). In both groups, patients measured their plasma TG before and 3 hours after each meal by Accutrend GCT (ROCHE, Mannheim, Germany). A subgroup of patients (n = 18) also performed TG determinations 2 hours after the test meals. The 3-hour TG increments were not significantly different between the different test meals (f = 0.671; P =.52); instead, the TG increments induced by fat supplements were significantly higher than those induced by the CHO supplements (f = 14.31; P =.0001). Similar results were also obtained 2 hours after the test meals. In conclusion, this study shows that the 2- and 3-hour TG responses to fat are higher compared with that induced by carbohydrate. This point, especially if confirmed by experiments with more frequent after meal measurements and of longer duration, should be taken into account in defining the best dietary approach to lower plasma TG levels throughout the whole day. PMID- 15131768 TI - Insulin secretion and incretin hormones after oral glucose in non-obese subjects with impaired glucose tolerance. AB - Subjects with impaired glucose tolerance (IGT) are usually overweight and exhibit insulin resistance with a defective compensation of insulin secretion. In this study, we sought to establish the interrelation between insulin secretion and insulin sensitivity after oral glucose in non-obese subjects with IGT and we also examined this interrelation in relation to the 2 main incretins, glucagon-like peptide (GLP-1) and gastric inhibitory polypeptide (GIP). To that end, 13 women with IGT and 17 women with normal glucose tolerance (NGT) underwent an oral glucose tolerance test (OGTT) with measurements of glucose, insulin, C-peptide, GLP-1, and GIP. Insulin secretion (TIS) and insulin sensitivity (OGIS) were assessed using models describing the relationship between glucose, insulin and C peptide data. These models allowed estimation also of the hepatic extraction of insulin. The age (54.2 +/- 9.7 [mean +/- SD] years) and body mass index (BMI; 26.0 +/- 4.0 kg/m(2)) did not differ between the groups. Subjects with IGT displayed lower TIS during the initial 30 minutes after oral glucose (0.97 +/- 0.17 [mean +/- SEM] v 1.75 +/- 0.23 nmol/L in NGT; P =.018) and lower OGIS (397 +/- 21 v 463 +/- 12 mL/min/m(2); P =.005). The incremental 30-minute TIS times OGIS (reflecting insulin secretion in relation to insulin sensitivity) was significantly reduced in IGT (359 +/- 51 v 774 +/- 91 nmol/min/m(2), P =.001). This measure correlated inversely to the 2-hour glucose level (r = -0.71; P <.001). In contrast, TIS over the whole 180-minute period was higher in IGT (26.2 +/- 2.4 v 20.0 +/- 2.0 nmol/L; P =.035). Hepatic insulin extraction correlated linearly with OGIS (r = 0.71; P <.001), but was not significantly different between the groups although there was a trend with lower extraction in IGT (P =.055). Plasma levels of GLP-1 and GIP increased after oral glucose. Total secretion of these incretin hormones during the 3-hour test did not differ between the 2 groups. However, the 30-minute increase in GLP-1 concentrations was lower in IGT than in NGT (P =.036). We conclude that also in non-obese subjects with IGT, when adiposity is controlled for in relation to NGT, defective early insulin secretion after oral glucose is a key factor. This defective beta-cell function is associated with, and may be caused by, a reduced early GLP-1 response. PMID- 15131769 TI - Comparison of proliferation and differentiation capacity of human adipocyte precursor cells from the omental and subcutaneous adipose tissue depot of obese subjects. AB - Upper body obesity is characterized by an expansion of the visceral adipose tissue and is associated with an increased susceptibility for type 2 diabetes and cardiovascular disease. In order to get a better understanding of the regulation of body fat distribution, the aim of the present study was to compare adipocyte development between the omental and subcutaneous adipose tissue region in obese subjects. Therefore, the proliferation and differentiation capacity in primary cultures of adipose tissue-derived stromal cells were compared between the 2 depots in a group of 29 obese individuals, of which 21 were women. Proliferation of the cells was stimulated using fetal calf serum (FCS) and assessed by counting the cell number in the culture dishes. Differentiation of preadipocytes was assessed in parallel by morphological criteria and determination of glycerol-3 phosphate dehydrogenase (GPDH) after stimulation by standardized adipogenic conditions. Stromal cells from the subcutaneous adipose tissue region proliferated faster (doubling time, 4 +/- 1 days) than those from the omental region (doubling time, 5 +/- 1 days), whereas there was no regional difference in adipose differentiation with any of the adipogenic media. The same findings were observed when men were excluded from the analysis. Interestingly, there were more endothelial cells in the cultures from the omental tissue as compared to those from the subcutaneous tissue, but there was no correlation between endothelial cell contamination and proliferation capacity, suggesting that the regional difference in proliferation capacity was not due to regional differences in the amount of endothelial cells. In addition, we found a negative correlation between donor age and proliferation of subcutaneous cells but not of omental cells, possibly explaining the greater capacity for adipose tissue growth in the omental as compared to the subcutaneous depot with aging. In conclusion, there may exist regional differences in adipose tissue growth with regard to proliferation capacity, whereas there are apparently no significant differences in in vitro differentiation capacity between subcutaneous and omental preadipocytes. PMID- 15131770 TI - Association of serum sialic acid with cardiovascular metabolic risk factors in Kuwaiti children and adolescents with type 1 diabetes. AB - The aim of the present study was to investigate the relation of serum total sialic acid (TSA) concentrations with cardiovascular metabolic risk factors in Kuwaiti children and adolescents with uncomplicated type 1 diabetes. This case control study included 150 (57 males and 93 females) type 1 diabetic children aged 6 to 18 years matched by age and sex to 150 nondiabetic children as controls. Measured variables included weight, height, systolic, diastolic blood pressure, and biochemical variables: blood glucose, glycated hemoglobin (HbA(1C)), triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL), apolipoproteins (apo) A1 and B, and urine microalbumin. There was no significant difference between mean serum TSA of the type 1 diabetic children (671.0 mg/L) and their controls (663.7 mg/L). In diabetic children, mean serum TSA was significantly higher in females (699.1 mg/L) than in males (625.2 mg/L) (P =.003). Significant correlations were found between serum TSA and the cardiovascular risk factors TC (P =.002), TG (P <.001), and apo B (P =.008). TSA mean level was significantly higher in diabetic children with poor glycemic control (HbA(1C) > 9.0%; P =.015), raised TC (P =.013), raised TG (P =.014), and in children with family history of cardiovascular disease (CVD; P =.02). In conclusion, the study suggests that serum TSA levels were not elevated in young type 1 diabetic children as compared with controls. The study also confirmed significant correlation of TSA concentrations with CVD risk factors TC, TG, and apo B, and as such serum TSA may be considered as a marker for CVD risk, especially in diabetic patients. A long-term prospective study is recommended to ascertain the longitudinal relationship of serum TSA with the adverse metabolic changes in type 1 diabetic children as complications prevail. PMID- 15131771 TI - The contribution of cardiorespiratory fitness and visceral fat to risk factors in Japanese patients with impaired glucose tolerance and type 2 diabetes mellitus. AB - It is still unclear as to how cardiorespiratory fitness and visceral fat accumulation contribute to coronary heart disease (CHD) risk factors in patients with diabetes mellitus. The purpose of the present study was to investigate whether cardiorespiratory fitness contributes to such risk factors independently of visceral fat accumulation. Two hundred Japanese patients (137 men and 63 women, aged 22 to 81 years) with impaired glucose tolerance (IGT) and type 2 diabetes mellitus (type 2 DM) without any intervention and pharmacological therapy participated in a cross-sectional study. The levels of fasting insulin, triglyceride (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and resting blood pressure were assessed. Maximal oxygen uptake (V.o(2max)), an index of cardiorespiratory fitness, was predicted by a graded exercise test using a cycle ergometer. Visceral fat area (VFA) was measured by computed tomography scan. The criteria for abnormalities of the risk factors were determined according to the standard values for Japanese. All subjects were divided equally into the following 3 groups according to their fitness level: low fit (V.o(2max) < 32 mL/kg/min in men, V.o(2max) < 26 mL/kg/min in women), mid-fit (32 < or = V.o(2max) < 36 in men, 26 < or = V.o(2max) < 30 in women), and high fit (V.o(2max) > or = 36 in men, V.o(2max) > or = 30 in women). The association between fitness level and the prevalence of abnormal values for these parameters was analyzed by a multiple logistic regression model adjusted for age and VFA. The odds ratio (OR) and 95% confidence interval (CI) for the prevalence of hyperinsulinemia were significantly lower in the mid-fit (OR = 0.35, 95% CI, 0.16 to 0.78) and in the high-fit groups (OR = 0.40, 95% CI, 0.16 to 0.98) compared with the low-fit group. In addition, ORs for the prevalence of low HDL-C in the mid-fit and high-fit groups were significantly lower (OR = 0.35, 95% CI, 0.14 to 0.86; and OR = 0.19; 95% CI, 0.08 to 0.60, respectively) than in the low-fit group. These results suggested that cardiorespiratory fitness might be one of the predictors of metabolic abnormalities, especially in patients with hyperinsulinemia and low HDL-C, independent of visceral fat accumulation in Japanese patients with IGT and type 2 DM. PMID- 15131772 TI - Association of Ob-R gene polymorphism and insulin resistance in Japanese men. AB - Leptin and its receptors are known to play a role in glucose metabolism. We succeeded in cloning human Ob-R cDNA and revealed 7 single nucleotide polymorphisms (SNPs) (Lys109Arg, Arg223Gln, Ser343Ser, Ser492Thr, Lys656Asn, Ala976Asp, and Pro1019Pro) in the coding region of Ob-Rb. Although these 7 SNPs were not associated with an obese phenotype, several studies have reported that some of them were associated with impaired glucose metabolism. To clarify whether the Arg223Gln and A3057G (Pro1019Pro) polymorphisms influence glucose metabolism in Japanese, 696 Japanese men were genotyped. Individually, the Arg223Gln and the A3057G polymorphisms were not associated with the glucose metabolic parameters. No associations were found between haplotype and clinical parameters. However, in 327 subjects with normal glucose tolerance (NGT), the subjects with Arg/Gln or Gln/Gln + A/A haplotype showed significantly higher serum insulin levels and homeostasis model assessment (HOMA) index than those with Arg/Arg + A/A haplotype and Arg/Gln or Gln/Gln + A/G or G/G haplotype. The subjects with Arg/Gln or Gln/Gln + A/A haplotype showed a significantly lower fasting glucose to insulin (GI) ratio than those with Arg/Arg + A/A haplotype. These results suggest that the Ob-R gene may serve as a modifier gene for insulin resistance in Japanese men. PMID- 15131773 TI - Inflexibility of energy substrate oxidation in type 1 diabetic patients. AB - Obese, insulin-resistant patients have been shown to have metabolic inflexibility. The goal of this study was to examine the effect of insulin administration on energy metabolism in lean, type 1 diabetic (DM1) patients. Eleven DM1 patients without vascular complications and 11 healthy controls (C) were examined. We performed a 2-step hyperinsulinemic euglycemic clamp (240 minutes; period 1: 1 mU. kg(-1). min(-1) and period 2: 10 mU. kg(-1). min(-1)) combined with indirect calorimetry during basal period B (B, -45 to 0 minutes), period 1, and period 2 of the clamp. The metabolic clearance rates of glucose (MCR) were lower in DM1 compared with C in period 1 (12.54 +/- 3.38 v 17.41 +/- 6.18 mL. kg(-1). min(-1); P <.02), as well as in period 2 (21.63 +/- 6.47 v 26.61 +/- 4.45 mL. kg(-1). min(-1); P <.05). Basal respiratory quotient (RQ) was lower in DM1 compared with C (0.72 +/- 0.04 v 0.75 +/- 0.04; P <.03). Insulin administration was accompanied by an increase in RQ in both groups, which was lower in DM1 compared with C (period 1: +0.09 +/- 0.04 v +0.11 +/- 0.07; P <.001; period 2: +0.13 +/- 0.04 v +0.16 +/- 0.04; P <.001). Glucose oxidation did not differ between the groups in period B; however, it was lower in DM1 compared with C in periods 1 (1.17 +/- 0.67 v 3.28 +/- 1.11 mg. kg(-1). min(-1); P <.003); and 2 (2.10 +/- 0.64 v 3.28 +/- 0.93 mg. kg(-1). min(-1); P <.009). Lipid oxidation was higher in DM1 in all periods compared with C; period B (3.28 +/- 0.77 v 1.16 +/- 0.55 mg. kg(-1). min(-1); P <.001), period 1 (1.10 +/- 0.41 v 0.67 +/- 0.54 mg. kg(-1). min(-1); P <.05), and period 2 (0.99 +/- 0.29 v 0.52 +/- 0.58 mg. kg( 1). min(-1); P <.01). The groups did not differ in protein oxidation. In conclusion, DM1 patients with secondary insulin resistance (IR) are characterized by metabolic inflexibility manifesting itself by smaller increases in RQ and glucose oxidation after insulin administration during the euglycemic clamp. PMID- 15131774 TI - Magnesium metabolism in mice selected for high and low erythrocyte magnesium levels. AB - A genetic control of blood magnesium (Mg) levels has been suggested. To investigate the mechanisms and the biologic significance of this genetic regulation, a mouse model, ie, mice selected for low magnesium level (MGL) and high magnesium level (MGH), was developed. The purpose of this study was to explore the Mg status and Mg metabolism in female MGL and MGH mice. We observed that MGL mice had reduced total and ionized plasma Mg, lower erythrocyte Mg, lower tibia, and kidney Mg levels. In contrast, total urinary Mg and (25)Mg levels were significantly higher in MGL mice. MGL mice had smaller total Mg exchangeable pool masses compared with MGH, and fractional transport rates of Mg (exchange constant) were different. In vitro (25)Mg enrichments in erythrocytes from MGL mice were significantly lower. Moreover, Mg efflux from erythrocytes was significantly higher in MGL. In conclusion, this work demonstrates that MGL mice present lower body stores of Mg than MGH mice and lower body Mg retention. This is confirmed at a cellular level by a lower enrichment of (25)Mg in erythrocytes. The lower retention of Mg by MGL erythrocyte in comparison to MGH appears to be partly due to a higher Mg efflux in MGL erythrocyte. It can be hypothesized that a genetic factor that modulates Na(+)/Mg(2+) exchanger activity may be important in the regulation of Mg metabolism. Further investigations on the mechanisms responsible for differences in Mg retention between MGL and MGH mice could contribute to a better understanding of the genetic regulation of cellular Mg. PMID- 15131775 TI - Partial androgen deficiency in aging type 2 diabetic men and its relationship to glycemic control. AB - Aging in the male is associated with both a higher incidence of type 2 diabetes and hypogonadism. However, little information is available about the complex of symptoms and hormonal changes related to partial androgen deficiency in aging (called andropause) in type 2 diabetic men. Here, for the first time, we used a combination of clinical and hormonal criteria to define andropause and to analyze the relationships between the androgen environment and glucose metabolism in 55 type 2 diabetic men (63.6 +/- 7.9 years, mean +/- SD). Low plasma levels of total testosterone (< or =3.4 ng/mL) and free testosterone (< or =11 pg/mL) were found in 20% and 54.5%, respectively, of the diabetic men. The fraction of diabetic men with subnormal levels of total testosterone increased with aging: 14.2% (50 to 59 years), 17.4% (60 to 69 years) and 36% (> 70 years). The corresponding figures for subnormal values of free testosterone were 38%, 69.6%, and 54.5%, respectively. In the whole group of type 2 diabetic men, no significant linear correlations between total or free testosterone with fasting plasma glucose, insulin, C-peptide, or fructosamine values could be established. Total testosterone was positively correlated with glycosylated haemoglobin (HbA(1c)) levels (r =.322, P =.01). Although fasting plasma glucose was marginally higher in aging type 2 diabetic patients with andropause than in those without andropause (162 +/- 6.9 v 139 +/- 8.9, mean +/- SEM, P =.05), there were no differences between both subgroups for plasma fasting insulin, C-peptide, fructosamine, or HbA(1c) levels. Replacement therapy (150 mg intramuscular [IM] of enanthate of testosterone every 14 days for 6 months) was applied in 10 type 2 diabetic men with clinical features of andropause associated with subnormal concentrations of serum testosterone. The treatment induced significant increases in total plasma testosterone (baseline: 3.9 +/- 0.3; at 6 months: 7.1 +/- 0.9 ng/mL, mean +/- SEM, P =.003) and free testosterone (baseline: 9.3 +/- 0.6; at 6 months 17.6 +/- 2.4 pg/mL, P =.003), but had a neutral effect on overall glycemic control. These data show a high prevalence of andropause in aging type 2 diabetic men and suggest that the endogenous androgen environment, as well as correction of the partial androgen deficiency, do not have a meaningful effect on glycemic control. PMID- 15131776 TI - Plasma nitrate/nitrite response to an oral glucose load and the effect of endurance training. AB - To assess the role of circulating nitric oxide (NO) production in glucose homeostasis, plasma nitrate/nitrite (NO(x)) was assessed during oral glucose tolerance tests (OGTTs) on 64 sedentary subjects and in a subset 40 subjects before and after 6 months of endurance exercise training. NO(x) decreased with the oral glucose load (P 40 IU/L or ALT >35 IU/L) who were prescribed a statin; cohort 2: 1437 hyperlipidemic patients with normal transaminases who were prescribed a statin; and cohort 3: 2245 patients with elevated liver enzymes but who were not prescribed a statin. The effect of statins on liver biochemistries was assessed over a 6-month period after statins were prescribed. Elevations in liver biochemistries during follow-up were categorized into mild-moderate or severe based on predefined criteria. RESULTS: The incidence of mild-moderate elevations and severe elevations in liver biochemistries in cohort 1 were 4.7% and 0.6%, respectively. Compared with cohort 1, individuals in cohort 2 had lower incidence of mild-moderate elevations (1.9%, P = 0.002) but not severe elevations (0.2%, P = 0.2). However, between cohorts 1 and 3, there were no differences in the incidence of mild-moderate elevations (4.7% vs. 6.4%, respectively, P = 0.2) or severe elevations (0.6% vs. 0.4%, respectively, P = 0.6). Statin discontinuation during the follow-up was similar between cohorts 1 and 2 (11.1% vs. 10.7%, respectively, P = 0.8). CONCLUSIONS: These data suggest that individuals with elevated baseline liver enzymes do not have higher risk for hepatotoxicity from statins. PMID- 15131790 TI - The effect of alcohol consumption on the prevalence of iron overload, iron deficiency, and iron deficiency anemia. AB - BACKGROUND & AIMS: Our aim was to investigate the relationship between alcohol consumption and iron overload, iron deficiency, or iron deficiency anemia in the U.S. population. METHODS: Adult participants of the Third National Health and Nutrition Examination Survey who did not consume alcohol (n = 8839) were compared with participants who consumed < or =1 (n = 4976), >1 to < or =2 (n = 1153), or >2 (n = 915) alcoholic drinks/day during the preceding 12 months. We examined the following markers of iron overload: elevated serum transferrin-iron saturation (TS) level (>45%, >50%, and >60%), elevated serum ferritin level (>300, >400, >500, and >600 ng/mL), and combinations of both elevated serum TS and ferritin levels. Iron deficiency was defined as the presence of at least 2 of the following: serum ferritin level <12 ng/mL, serum TS level <15%, and erythrocyte protoporphyrin level >1.24 micromol/L. Iron deficiency anemia was defined as the presence of both iron deficiency and anemia. RESULTS: Compared with nondrinkers, the prevalence of all markers of iron overload was significantly elevated among those who consumed >2 alcoholic drinks/day after adjusting for potential confounders. Consumption of any amount of alcohol was associated with a 40% reduction in the risk of iron deficiency anemia. CONCLUSIONS: Consumption of up to 2 alcoholic drinks/day seems to be associated with reduced risk of iron deficiency and iron deficiency anemia without a concomitant increase in the risk of iron overload. Consumption of >2 alcoholic drinks/day is associated with a significant elevation in the risk of iron overload. PMID- 15131791 TI - Epoetin alfa maintains ribavirin dose in HCV-infected patients: a prospective, double-blind, randomized controlled study. AB - BACKGROUND & AIMS: Combination therapy with interferon alpha (IFN-alpha) and ribavirin (RBV) or pegylated IFN-alpha (PEG-IFN-alpha)/RBV for chronic hepatitis C virus (HCV) infection often causes anemia, prompting RBV dose reduction/discontinuation. This study assessed whether epoetin alfa could maintain RBV dose, improve quality of life (QOL), and increase hemoglobin (Hb) in anemic HCV-infected patients. METHODS: HCV-infected patients (n = 185) on combination therapy who developed anemia (Hb < or = 12 g/dL) were randomized into a U. S. multicenter, placebo-controlled, clinical trial of epoetin alfa, 40,000 U subcutaneously, once weekly vs. matching placebo. The study design used an 8 week, double-blind phase (DBP) followed by an 8-week, open-label phase (OLP), in which placebo patients were crossed over to epoetin alfa. RESULTS: At the end of the DBP, RBV doses were maintained in 88% of patients receiving epoetin alfa vs. 60% of patients receiving placebo (P < 0.001). Mean QOL scores at the end of the DBP improved significantly on all domains of the Linear Analog Scale Assessment (LASA) and on 7 of the 8 domains of the Short Form-36, version 2 (SF-36v2). Mean Hb increased by 2.2 +/- 1.3 g/dL (epoetin alfa) and by 0.1 +/- 1.0 g/dL (placebo) in the DBP (P < 0.001). Similar results were demonstrated in patients who switched from placebo to epoetin alfa in the OLP. Epoetin alfa was well tolerated; the most common adverse effects were headache and nausea. CONCLUSIONS: Epoetin alfa maintained RBV dose and improved QOL and Hb in anemic HCV-infected patients receiving combination therapy. PMID- 15131792 TI - Association of antioxidant enzyme gene polymorphisms and glutathione status with severe acute pancreatitis. AB - BACKGROUND & AIMS: Genetic variations in antioxidant metabolism may explain varying biological responses to acute pancreatitis (AP). We studied the contribution of oxidative stress to the pathogenesis of severe pancreatitis by examining the prevalence of functional gene polymorphisms of antioxidant enzymes and evidence of heightened oxidative stress. METHODS: DNA from 320 patients with AP (90 severe) and 263 controls was genotyped for glutathione S-transferase (Mu-1 [M-1], theta-1 [T-1], and pi-1 [P-1: Ile-105Val]), manganese superoxide dismutase (Ala-9Val), and catalase (C-260T) polymorphisms. Erythrocyte reduced glutathione (GSH) concentration was determined 24 and 72 hours after the onset of pain in 46 patients (11 severe). Disease severity was assessed using Atlanta clinical criteria, Acute Physiology Scores (APS), and peak serum C-reactive protein levels. RESULTS: The functional GSTT-1*A genotype was more prevalent in severe (96%) compared with mild attacks of AP (78%; odds ratio [OR], 5.9; 95% confidence interval [CI ], 2-17; P < 0.0001) and controls (76%; OR, 6.6; 95% CI, 2.3-18.7; P < 0.0001). Compared with null genotype, GSTT-1*A was associated with higher peak C-reactive protein levels (184 vs. 94 g/dL; P = 0.0005) and APS (24 hours, P = 0.04; 48 hours, P = 0.015). Reduced glutathione (GSH) at 24 hours was lower in mild (median, 382 nmol/g) and severe attacks (median, 407 nmol/g) compared with controls (median, 3685 nmol/g; P < 0.001). Levels increased at 72 hours in mild (P = 0.012) but not severe attacks and inversely correlated with APS (r = -0.49; P = 0.04). CONCLUSIONS: The functional GSTT-1*A genotype was associated with severe attacks of pancreatitis. Heightened oxidative stress characterized by glutathione depletion may be of importance in mediating the progression from mild to severe pancreatitis. PMID- 15131793 TI - Telangiectatic focal nodular hyperplasia: a variant of hepatocellular adenoma. AB - BACKGROUND & AIMS: "Telangiectatic focal nodular hyperplasia" designate atypical lesions considered as variants of focal nodular hyperplasia (FNH). However, because "telangiectatic FNH" share several morphologic patterns with hepatocellular adenomas, classification of such lesions deserve further clarification. Therefore, the aim of the present study was to reconsider the classification of telangiectatic FNH with the help of a molecular approach. METHODS: Ten telangiectatic FNH, 6 typical FNH, and 6 hepatocellular adenomas were studied. DNA, RNA, and protein from each lesion were extracted. Clonality was assessed by the study of the X chromosome inactivation pattern (HUMARA assay). Angiopoietin (ANGPT-1 and ANGPT-2) mRNA, genes the expression of which is typically modified in FNH, were quantified by a real-time RT-PCR procedure. Protein profiles were analyzed by SELDI-TOF PROTEINCHIP (Cyphergen Biosystem, Inc., Fremont, CA) technology. RESULTS: Although all informative cases of FNH (5 of 6) and hepatocellular adenomas (6 of 6) were polyclonal and monoclonal, respectively, clonal analysis showed a nonrandom pattern of X chromosome inactivation consistent with a monoclonal lesion in 6 of 8 cases of telangiectatic FNH. The mean value of the ANGPT-1/ANGPT-2 mRNA ratio was 21.4 in FNH, 2.6 in adenomas, and 2.1 in telangiectatic FNH (P 10-fold and >600 fold elevated in the adenocarcinoma cells as compared with normal gastric/intestinal cells and normal diploid fibroblasts, respectively. Telomeres were short, being less than 4 kilobase pair in both tumor cell lines. Exposure to PPA effectively inhibited telomerase activity and shortened telomeres. PPA also arrested cell proliferation and reduced colony number and size after a lag period of about 10 cell generations, consistent with the attrition of telomeres. The growth arrest was not due to senescence but was due to apoptosis. Expression analysis of the cells following PPA treatment did not show significant change in the expression of genes involved in cell-cycle proliferation and apoptosis. Exposure to PPA had no effect on proliferative potential of normal intestinal cells. CONCLUSIONS: We conclude that telomerase inhibition by PPA induces cell growth arrest in BEAC cells and demonstrate the potential of telomerase inhibitors in chemoprevention and treatment of Barrett's-associated esophageal adenocarcinoma. PMID- 15131796 TI - The expression and function of costimulatory molecules B7H and B7-H1 on colonic epithelial cells. AB - BACKGROUND & AIMS: Previous studies have suggested that intestinal epithelial cells (IECs) may function as antigen-presenting cells for CD4+ and CD8+ T cells. However, these cells fail to express conventional costimulatory molecules (CD80, CD86), leading to the possibility that antigen presented by normal IECs could result in anergy. Other members of the B7 family have recently been identified. B7h interacts with inducible costimulator (ICOS) on T cells and provides a positive signal, whereas B7-H1 and B7-DC interact with PD-1 and transmit an inhibitory signal. Our aim was to determine whether IECs express novel B7 family members and whether these molecules play a role in IEC:T-cell interactions. METHODS: B7h and B7-H1 expression was assessed in isolated IECs and IEC lines. The functional role of B7h and B7-H1 in the interaction between IECs and T cells was assessed in coculture experiments using purified anti-B7h or B7-H1 monoclonal antibodies (mAbs), B7h immunoglobulin (Ig), or B7-H1 fusion proteins. RESULTS: B7h and B7-H1 messenger RNA was detected in IEC lines and IECs from healthy controls and patients with inflammatory bowel disease (IBD). IECs from patients with IBD but not healthy controls expressed B7h and B7-H1 protein on their surface. Proliferation of IEC-stimulated T cells was inhibited only by B7h immunoglobulin treatment, whereas interferon gamma secretion in these cocultures was inhibited by both anti-B7h mAb and B7h Ig. No difference was seen between IBD or normal IEC populations. CONCLUSIONS: These data suggest that the B7h-ICOS costimulatory pathway may be important in IEC:T-cell interactions. PMID- 15131797 TI - DNA from probiotic bacteria modulates murine and human epithelial and immune function. AB - BACKGROUND & AIMS: The intestinal epithelium must discriminate between pathogenic and nonpathogenic bacteria and respond accordingly. The aim of this study was to examine whether bacterial DNA can serve as the molecular basis for bacterial recognition. METHODS: HT-29 monolayers were treated with various bacterial DNA and interleukin (IL)-8 secretion measured by enzyme-linked immunosorbent assay, nuclear factor kappaB activation by electrophoretic mobility shift assay and reporter assays, and IkappaB levels by Western blotting. Cytokine secretion in response to bacterial DNA was measured in murine colonic segments and splenocytes. IL-10-deficient mice were fed DNA from VSL probiotic compound daily for 2 weeks. Colons were removed and analyzed for cytokine production and inflammation. RESULTS: HT-29 cells responded with IL-8 secretion to bacterial DNA in a differential manner. In the presence of proinflammatory stimuli, VSL3 DNA inhibited IL-8 secretion, reduced p38 mitogen-activated protein kinase activation, delayed nuclear factor kappaB activation, stabilized levels of IkappaB, and inhibited proteasome function. VSL3 DNA inhibited colonic interferon (IFN)-gamma secretion in mouse colons and also attenuated a Bacteroides vulgatus induced IFN-gamma release from murine splenocytes. In mice, VSL3 DNA attenuated a systemic release of tumor necrosis factor alpha in response to Escherichia coli DNA injection. Treatment of IL-10-deficient mice with oral VSL3 DNA resulted in a reduction in mucosal secretion of tumor necrosis factor alpha and IFN-gamma and an improvement in histologic disease. CONCLUSIONS: DNA from probiotic bacteria can limit epithelial proinflammatory responses in vivo and in vitro. Systemic and oral administration of VSL3 DNA ameliorates inflammatory responses. PMID- 15131798 TI - Disruption of beta-catenin pathway or genomic instability define two distinct categories of liver cancer in transgenic mice. AB - BACKGROUND & AIMS: Human liver cancer can be divided into 2 categories that are characterized by activation of beta-catenin and genomic instability. Here we investigate whether similar categories exist among 5 transgenic models of liver cancer, including c-myc, transforming growth factor-alpha, E2F-1, c myc/transforming growth factor-alpha, and c-myc/E2F-1 mice. METHODS: The random amplified polymorphic DNA method was used to assess the overall genomic instability, and chromosomal loci affected by genomic alterations were determined by microsatellite analysis. beta-Catenin mutations and deletions were analyzed by polymerase chain reaction and sequencing screening. Cellular localization of beta catenin and expression of alpha-fetoprotein, a prognostic marker of hepatocellular carcinoma, were investigated by immunohistochemistry. RESULTS: Liver tumors from the transgenic mice could be divided into 2 broad categories characterized by extensive genomic instability (exemplified by the c myc/transforming growth factor-alpha mouse) and activation of beta-catenin (exemplified by the c-myc/E2F-1 mouse). The c-myc/transforming growth factor alpha tumors displayed extensive genomic instability with recurrent loss of heterozygosity at chromosomes 1, 2, 4, 6, 7, 9, 12, 14, and X and a low rate of beta-catenin activation. The genomic instability was evident from the early dysplastic stage and occurred concomitantly with increased expression of alpha fetoprotein. The c-myc/E2F-1 tumors were characterized by a high frequency of beta-catenin activation in the presence of a relatively stable genome and low alpha-fetoprotein levels. CONCLUSIONS: We have identified 2 prototype experimental models, i.e., c-myc/transforming growth factor-alpha and c-myc/E2F-1 mice, for the 2 categories of human hepatocellular carcinoma characterized by genomic instability and beta-catenin activation, respectively. These mouse models will assist in the elucidation of the molecular basis of human hepatocellular carcinoma. PMID- 15131799 TI - Activated natural killer T cells induce liver injury by Fas and tumor necrosis factor-alpha during alcohol consumption. AB - BACKGROUND & AIMS: Chronic alcohol abuse induces liver injury and increases the severity of viral hepatitis, but the precise mechanisms responsible are not well understood. In particular, little is known about the role of natural killer T cells in alcohol-induced liver injury. Natural killer T cells are mediators of important regulator and effector functions making use of Fas and tumor necrosis factor (TNF)-alpha in apoptosis induction. This report analyzes the role of natural killer T cells, Fas, and TNF-alpha in a model of chronic alcohol consumption. METHODS: Mice fed alcohol by intragastric tube were assayed for serum alanine aminotransferase values, liver histology, and liver mononuclear cells before and after activation of natural killer T cells by ligand alpha galactosylceramide. RESULTS: In alcohol-consuming animals, liver natural killer T cells increase, and further activation by alpha-galactosylceramide causes lethal liver injury. This is explained by alcohol-induced hepatocyte sensitization to cell-mediated lysis, which develops concomitant to increased cytolytic activity of natural killer T cells. Natural killer T cell-mediated apoptosis proceeds by the Fas pathway, and Fas is essential for alcohol-associated liver injury. TNF alpha plays an additional role as a defect in TNF receptor-1 inhibits alcohol associated liver injury. Alcohol-fed natural killer T cell-deficient Jalpha281(-/ ) mice express a delay in alcohol-induced liver injury. CONCLUSIONS: Alcohol consumption induces an increase of natural killer T cells in the liver and a high sensitivity of hepatocytes to cell-mediated lysis. Stimulation of natural killer T cells during alcohol consumption induces serious liver injury by a mechanism that involves concomitant signals by Fas and tumor necrosis factor receptor-1 on alcohol-stressed hepatocytes. PMID- 15131800 TI - Multigenic control of hepatic iron loading in a murine model of hemochromatosis. AB - BACKGROUND & AIMS: Hereditary hemochromatosis is a common disorder of iron homeostasis characterized by increased dietary iron absorption and progressive iron accumulation, mainly in the liver. Most patients are homozygous for the C282Y mutation in the HFE gene. However, not all individuals carrying the hemochromatosis-predisposing genotype in the general population become iron loaded. Genetic modifiers have been shown to influence disease penetrance, but their number and chromosomal locations remain unknown, and their identification is hampered by complex interactions with environmental factors. To circumvent these difficulties, we used 2 strains of mice made deficient for the Hfe gene that strongly differ in their propensity to develop hepatic iron loading. METHODS: To localize the loci controlling hepatic iron loading in this murine model of hemochromatosis, we produced 1028 mice by an F2 intercross between the C57BL/6 and DBA/2 Hfe-deficient strains. We selected the 276 mice that contributed the most to the total linkage information for genotyping with 145 microsatellite markers. RESULTS: We mapped 4 modifier loci on chromosomes 7, 8, 11, and 12, with logarithm of odds scores of 14.47, 12.96, 6.04, and 6.72, respectively, in regions containing several genes recently shown to exert important roles in the regulation of iron metabolism. CONCLUSIONS: Our data provide a clear demonstration of the polygenic pattern of hepatic iron loading inheritance in Hfe-deficient mice. Examination of candidate genes residing at the loci identified in this study and genetic analysis of the syntenic chromosomal regions in humans may provide important insight into the heterogeneous disease presentation observed among HFE C282Y homozygotes. PMID- 15131801 TI - Chronic hepatitis C with normal aminotransferase levels. PMID- 15131802 TI - Liver biopsy discloses a new apolipoprotein A-I hereditary amyloidosis in several unrelated Italian families. AB - BACKGROUND & AIMS: Hereditary systemic amyloidoses are autosomal dominant, late onset disorders caused by mutations in the genes for a group of plasma proteins including transthyretin, lysozyme, fibrinogen Aalpha chain, gelsolin, apolipoprotein A-I, and apolipoprotein A-II. We investigated both phenotypic and genotypic aspects of apolipoprotein A-I amyloidosis unexpectedly disclosed by liver biopsy in 13 unrelated individuals with asymptomatic, persistent elevation of alkaline phosphatase and gamma-glutamyltransferase levels. METHODS: Immunoelectron microscopy was used for in situ characterization of amyloid deposits on liver biopsy specimens. Mutation analysis was performed by sequencing of the apolipoprotein A-I gene in all patients. Wild-type/variant apolipoprotein A-I ratio in plasma high-density lipoproteins was assessed by a peptide mass fingerprinting approach after purification of total apolipoprotein A-I of 2 patients. RESULTS: Family history was informative in 5 cases. Renal failure developed in 9 cases. Hypogonadism due to testicular involvement was observed. Amyloid fibrils specifically stained with anti-apolipoprotein A-I antibody. A novel (Leu75Pro) heterozygous mutation in the apolipoprotein A-I gene was present in affected individuals but not in controls. Variant apolipoprotein A-I was about 10% of the total protein in high-density lipoproteins. CONCLUSIONS: The high number of individuals with apparently sporadic disease might reflect widespread occurrence of this mutation in the population and a milder phenotype of this variant compared with other apolipoprotein A-I amyloidogenic mutants. These findings suggest that specific staining for amyloid should be performed on liver biopsy of individuals with asymptomatic chronic elevation of alkaline phosphatase and gamma-glutamyltransferase levels. PMID- 15131803 TI - Colorectal cancer chemoprevention--an overview of the science. AB - The development and dissemination of sophisticated detection technologies have recently exposed the high prevalence of preinvasive colorectal neoplasia in the adult U.S. population. Although cancer screening and surveillance provide opportunities for risk stratification, they achieve risk reduction only when coupled with effective interventions. This review surveys the lead compounds for colorectal cancer prevention and the measures by which they may be prioritized for clinical testing. Clinical trials remain the rate-limiting step in agent development, and novel trial designs are needed to hasten agent identification and testing for cancer prevention. Innovative research models include the nesting of prevention end points within cancer treatment trials and within trials testing promising preventive compounds intended for nononcologic indications. PMID- 15131804 TI - Digestive and liver diseases statistics, 2004. AB - BACKGROUND & AIMS: Digestive and liver diseases are associated with substantial morbidity and mortality in the United States. Statistics about the incidence, prevalence, mortality, and resource utilization of digestive and liver diseases in the United States may be cumbersome to obtain because they are scattered in multiple sources. These data may be useful for policy makers, grant applicants, and authors. METHODS: Data on the most common gastrointestinal and liver diseases were collected from large publicly available national databases. Information was collected on inpatient and outpatient gastrointestinal complaints and diagnoses, gastrointestinal cancers, and deaths from common liver diseases. RESULTS: The leading gastrointestinal complaint prompting an outpatient visit is abdominal pain, with 12.2 million annual visits, followed by diarrhea, nausea, and vomiting. Abdominal pain is the leading outpatient gastrointestinal diagnosis, accounting for 5.2 million visits annually, followed by gastroesophageal reflux disease, with 4.5 million visits. Gallstone disease is the most common inpatient diagnosis, with 262,411 hospitalizations and a median inpatient charge of USD$11,584. Colorectal cancer is the most common gastrointestinal cause of death and is the most common gastrointestinal cancer, with an incidence of 54 per 100,000. Among gastrointestinal cancers, primary liver cancer had the highest increase in incidence from 1992 to 2000. CONCLUSIONS: Gastrointestinal and liver diseases are associated with significant outpatient and inpatient healthcare utilization. Following trends in utilization is important for determining allocation of resources for health care and research. PMID- 15131805 TI - Our new President--Emmet B. Keeffe, M.D. PMID- 15131806 TI - American Gastroenterological Association medical position statement: Diagnosis and treatment of hemorrhoids. AB - This document presents the official recommendations of the American Gastroenterological Association (AGA) on Hemorrhoids. It was approved by the Clinical Practice Committee on January 8, 2004, and by the AGA Governing Board on February 13, 2004. PMID- 15131807 TI - American Gastroenterological Association technical review on the diagnosis and treatment of hemorrhoids. PMID- 15131808 TI - Quality control for flexible sigmoidoscopy: which polyps count? PMID- 15131809 TI - Are patients with elevated liver tests at increased risk of drug-induced liver injury? PMID- 15131810 TI - These FADs are here to stay--clinicopathological patterns of food allergic diseases. PMID- 15131811 TI - Endoscopic colorectal cancer screening--can supply meet demand? PMID- 15131814 TI - Beyond interferon for hepatitis C: living in the present while hoping for the future. PMID- 15131812 TI - Treatment of chronic HCV--should we stop treating nonresponders? PMID- 15131819 TI - Additional discussion on the safety of 6-mercaptopurine for childbearing patients with inflammatory bowel disease: a retrospective cohort study. PMID- 15131821 TI - Progression of flat low-grade dysplasia to advanced neoplasia in patients with ulcerative colitis. PMID- 15131823 TI - Low-grade dysplasia in flat mucosa in ulcerative colitis. PMID- 15131824 TI - Open questions concerning cyclosporine therapy in ulcerative colitis. PMID- 15131826 TI - A randomized controlled trial of biofeedback that does not have the power to conclude. PMID- 15131828 TI - Fundic gland polyps: not so trivial entity and worth evaluation. PMID- 15131829 TI - Absence of hepatitis C virus infection among patients with lymphoproliferative disorders in northern India. PMID- 15131831 TI - EUS FNA staging of esophageal cancer. PMID- 15131835 TI - Eicosanoids in cystic fibrosis. PMID- 15131836 TI - Postmenopausal osteoporosis and strontium ranelate. PMID- 15131837 TI - Risks of testosterone replacement. PMID- 15131838 TI - Risks of testosterone replacement. PMID- 15131839 TI - Risks of testosterone replacement. PMID- 15131840 TI - Parvovirus B19. PMID- 15131841 TI - Multidisciplinary management of lung cancer. PMID- 15131842 TI - Multidisciplinary management of lung cancer. PMID- 15131843 TI - Multidisciplinary management of lung cancer. PMID- 15131844 TI - Optical media: superluminal speed of information? PMID- 15131846 TI - "Headaches: assessment and management". PMID- 15131845 TI - ONA leads the fight for safe needles that save lives. PMID- 15131847 TI - Recruiting nurses from the Philippines to combat the nursing shortage in central Ohio. PMID- 15131848 TI - Are there some general rules concerning documentation? PMID- 15131849 TI - The physiologic basis of pain medications. AB - Certainly it is important for good nursing practice to know what medications are being prescribed, the desired therapeutic effects, and the anticipated undesirable side effects. Understanding the physiologic rationale for pain medications is, therefore, basic to good nursing care of persons with pain. Understanding the physiology of pain and rationale for use of medications also underscores the importance of a good pain assessment. Neuropathic pain often does not respond to traditional analgesics and the nurse's assessment will assist in identifying the type of pain a person is experiencing. A knowledge of how and why medications work to manage pain is also essential in order to provide good patient and family education. Misunderstandings can arise if individuals do not understand the rationale for their medications. For example, a patient who receives a prescription for desipramine may learn from his pharmacist that his new medication treats depression, leading to questions about the perception of his emotional status. Misinformation and confusion causes anxiety and can undermine trust in healthcare providers. Therefore, the rationale for all medications should be explained to patients. This is especially important when using medications that are not traditionally viewed as analgesics, such as tricyclic antidepressants and anticonvulsants. In most circumstances, pain can be very effectively treated. This requires a thorough initial assessment to determine the type and cause of pain, the careful selection of interventions, systematic evaluation of the effectiveness of interventions, and appropriate changes in the plan if optimal pain management is not achieved. The focus of this paper is on the rationale for use of various medications to manage pain. However, it is essential to remember that non-pharmacological interventions are very important in managing pain and must be included in a comprehensive plan of care for pain management. PMID- 15131850 TI - What every RN should know about a grievance. PMID- 15131851 TI - Nurses to adjust the infusion rates of epidural catheters for patients in labor. PMID- 15131852 TI - [Deep inferior epigastric perforator (DIEP) free flap in immediate breast reconstruction]. AB - OBJECTIVE: In order to facilitate psychological rehabilitation of the women who underwent modified radical mastectomy for breast cancer, the DIEP free flap was used for immediate breast reconstruction in a series of patients. We present a review of the surgical outcomes. METHODS: From December 2001 to January 2003, unilateral breast reconstruction was performed in 12 patients using the DIEP free flap at the same time of modified radical mastectomy in our department. The recipient vessels were the thoracodorsal artery and vein or the internal mammary artery and its venae concomitants. RESULTS: Of the 12 DIEP flaps, 1 was vascularized by a single perforator, 3 were vascularized by two perforators, 6 by three perforators, 1 by four an 1 by five perforators. One flap failed totally, likely due to previous multiple lower abdominal operations. The eleven reconstructed breasts achieved almost the same size and shape as the healthy sides. The time of patient getting out of bed ranged from 3 to 7 days and the mean hospital stay was 8.8 days. All patients were satisfied with the outcome. No complications were observed in the abdominal wall, including weakness, abdominal bulge or hernia. CONCLUSION: Immediate breast reconstruction facilitates the psychological rehabilitation and helps to avoid the dressing inconveniences resulted from total mastectomy. Free DIEP flap is a new and reliable technique for immediate breast reconstruction with autologous tissue. This flap offers the patients the same advantages as the TRAM flap and avoids its most important disadvantage of potential abdominal wall weakness, by preserving the continuity of the rectus abdominis muscle. The donor site morbidity is thus reduced and recovery is faster. The more complex nature of this procedure leads to increased operating time and requires more demanding surgical skills. PMID- 15131853 TI - [TRAM flap in conjunction with latissimus dorsi muscle flap for breast reconstruction]. AB - OBJECTIVE: To investigate a method to reconstruct the breast and repair the chest wall defects at the same time. METHODS: The operation procedure combined the transverse rectus abdominis myocutaneous (TRAM) flap with the latissimus dorsi muscle(LDM) flap for breast reconstruction and repair of chest wall defect. Two patients underwent delayed breast reconstruction using this technique. RESULTS: 8 flaps in the four patients survived completely. The aesthetic results were very good. CONCLUSION: This method can be used to reconstruct breast and repair the defect of chest wall at the same time, avoiding the disadvantage in the flap transfer of TRAM or LDM. PMID- 15131854 TI - [The complications of radiotherapy for breast cancer and the treatment for radiation ulcer]. AB - OBJECTIVE: To explore the effective treatment for chronic ulcer following radiotherapy for breast cancer and reveal the universality and severity of radiation-induced brachial plexus neuropathy. METHODS: The TRAM flap, the local expanded flap or the delayed skin flap were applied to repair the ulcer wounds in 16 patients. Electromyogram examinations were used to evaluate the radiation lesions of the brachial plexus. RESULTS: All the flaps survived successfully with satisfactory results except one, which sustained partial necrosis due to infection. Ten patients underwent regular electromyogram examinations, seven of them were found to have radiation-induced brachial plexus neuropathy. CONCLUSIONS: Radiation ulcer following radiotherapy for breast cancer is often concomitant with brachial plexus neuropathy. These injuries presented a chronically progressive and irreversible course. Application of the flaps that have adequate blood supply can reconstruct the wounds effectively. PMID- 15131855 TI - [Reduction mammaplasty with the superior-lateral dermo-glandular pedicle]. AB - OBJECTIVE: To introduce a new technique for reduction mammaplasty appropriate to moderate or heavy hypertrophic breast. METHODS: The superior-lateral dermo glandular pedicle flap including the nipple-areola complex was created. After the extra glandular tissue was removed, the superior-lateral dermo-glandular pedicle with the nipple-areola complex was rotated, adjusted, sculptured and fixed to the thoracic wall so as to fashion a breast with natural projection and proper shape. This method was used in 4 patients(8 breasts). RESULTS: The operation results were satisfactory without complications. The breasts maintained nice configuration and good function, with well lactating in one case. CONCLUSION: The technique of reduction mammaplasty with the superior-lateral dermo-glandular pedicle is a reasonable method to obtain nice breast configuration and good function. PMID- 15131856 TI - [Vertical mammaplasty using the skin wrinkling suture techniques]. AB - OBJECTIVE: To reduce the incision scar produced by reduction mammaplasty. METHODS: The periareola incision and the vertical incision below the areola were made when performing breast reduction. The incision in the submammary fold was avoided. The breast was suspended and moulded. The redundant skin and the long skin incision below the areola were sutured utilizing the skin wrinkling suture techniques. The wrinkling skin and incision became flat gradually due to the elasticity of the tissue itself. RESULTS: 13 cases have been treated with the techniques mentioned above. All incisions healed primarily and there were no complications such as hematoma, infection and necrosis of skin or areola. Of them, 3 patients with very large and ptotic breasts needed small corrective operations to excise the redundant skin that still existed 6 months postoperatively. The shape of the breasts and the vertical incision scars of the other patients were satisfactory. CONCLUSIONS: The vertical mammaplasty using the skin wrinkling suture techniques can be used in breast reduction of various sizes, owing to the advantages of consistent, good and stable result, simplicity to perform and grasp and no scar in the submammary fold. PMID- 15131857 TI - [Correction of congenital nipple inversion with bilateral rhombus deepithelialized subcutaneous pedicle skin flaps and continuous traction]. AB - OBJECTIVE: A new method for correction of nipple inversion was introduced to improve the surgical result. METHODS: In this procedure, two opposite rhombus incisions on both sides of the inverted nipple were designed within the range of areola. The rhombus skin was deepithelialized and two subcutaneous pedicle skin flaps were created. Then the shortened lactiferous ducts and fibrous bands beneath the nipple were released. The subcutaneous pedicle skin flaps on both sides of the inverted nipple were advanced through the tunnel beneath the nipple as the supporting tissue. After the operation, continuous traction of the nipple lasted two weeks at least. RESULTS: Since 1999, 16 patients with congenital bilateral nipple inversion have been treated. No complications were found in all the cases. Postoperative follow-up for 2-12 months showed that the appearance of the nipple was satisfactory. CONCLUSIONS: This method is simple, safe and effective for correction of congenital nipple inversion. The incision scar is indistinct, the natural look and good function of the nipple were reserved. PMID- 15131858 TI - [Vascular anatomy and clinical applications of the distally based superficial sural artery island flap]. AB - OBJECTIVE: To document the vascular anatomy of the distally based superficial sural artery flap and to study the vascular anastomoses between the superficial sural artery and the septocutaneous perforators of the peroneal artery. METHODS: Ten fresh human cadavers were injected with lead oxide, gelatin and water. Twenty lags were then dissected and an overall map of the cutaneous vasculature was constructed. Vascular communications between the superficial sural artery and the lowest septocutaneous perforator of the peroneal artery was evaluated to determine the cutaneous vascular territory of the superficial sural flap. The distally based superficial sural artery island flap was used in 26 cases. RESULTS: There is constant vascular anastomosis between the superficial sural artery and the lowest septocutaneous perforator of the peroneal artery. The 26 flaps survived uneventfully except for two of partial fat necrosis. CONCLUSION: The anatomic information enhances our understanding of flap design. PMID- 15131859 TI - [Anatomic study of perforator neurovascular bundles of the deep inferior epigastric perforator flap]. AB - OBJECTIVE: To investigate the anatomic characteristics of the perforator neurovascular bundles in the anterior abdominal wall for the anatomical basis of the sensate deep inferior epigastric perforator (DIEP) flap. METHODS: Abdominal wall dissection was performed on 9 embalmed female cadavers (18 sides). RESULTS: Most of the deep inferior epigastric perforators emerged at the anterior sheath of the rectus. There were averaged 17.5 perforators on each side, with the diameter equal to or larger than 0.5 mm in 7.8 perforators. The number of skin perforators was the greatest in the supero-medial area. 56.2 percent of vascular perforators traveled with the sensory branches while 80.9 percent of larger perforators (> or = 0.5 mm in diameter) run with the sensory branches. CONCLUSIONS: The lateral perforator neurovascular bundle was the first choice in the design of the sensate DIEP flap. PMID- 15131860 TI - [Repair of postburn hand using scarred skin and an abdominal flap in patients with severe burns]. AB - OBJECTIVE: To introduce a method for repairing and restoring hand function of the patients suffering from large surface area burn with scarce normal skin for reconstruction. METHODS: Seven patients (ten hands) were treated in our department from April 1994 to February 2001. The TBSA involved with second- or third-degree burns was 85%-96%. All the hands had severe scar contracture on the dorsum and lost most of their function. A scarred skin flap, based on the ulnar border of the hand was elevated integrally on the dorsum. A random-pattern abdominal flap at a less-scarred area was designed and elevated to cover the defect of the hand while the scarred skin flap of the hand was transferred to the donor site of the abdominal flap. RESULTS: All patients were followed for 0.5 to 4 years postoperatively. The range of motion of the metacarpophalangeal joint and the space capacity of the first web were greatly improved. All patients regained self-care ability. CONCLUSIONS: This method is simple and has satisfactory results. Under the circumstances where normal skin was not available for reconstruction, the function of the burned hand could be greatly improved by this method. PMID- 15131861 TI - [A clinical study on the extended reverse digital artery island flap]. AB - OBJECTIVE: To investigate the feasibility and clinical effect of the extended reverse digital artery island flap including dorsal digital nerve. METHODS: Since 2001, 7 cases of pulp and digital palm defects were repaired by the extended reverse digital artery island flap including dorsal digital nerve. RESULTS: All the flaps survived. Follow-up showed satisfactory results functionally and cosmetically with 2-PD of 6-9 mm. CONCLUSIONS: The extended reverse digital artery island flap is a safe and feasible and an effective method for repair of finger pulp defects. PMID- 15131862 TI - [Repair of foot defect with a reverse fascial pedicled posterolateral lower leg flap]. AB - OBJECTIVE: To explore the applications of ipsilateral reverse fascial pedicled posterolateral lower leg flap in foot defect repair. METHODS: A fasciocutaneous flap was designed over the gastrocnemius muscle to repair the defects in the foot or the lower third of the leg. The flap was based on a perforator branch between the lateral malleolus and the Achilles tendon, coming from the vessel network of the ankle as well as the deep fascia. RESULTS: This method was used in 6 cases, including 2 children of 5-6 years old. All the flaps survived without necrosis and venous congestion. CONCLUSIONS: The operation is simple and a large flap over the whole gastrocnemius muscle can be raised without sacrifice of a main blood vessel. It is an excellent method for repair of the defects in the heel, medial and lateral malleolus, dorsum pedis and the lower third of the leg. PMID- 15131863 TI - [Investigation of the influence of nitric oxide on the survival of a random pattern skin flap]. AB - OBJECTIVE: To investigate the changes of nitric oxide concentration in the distal portion of a random pattern skin flap and the influence of the exogenous L arginine on the survival of the random pattern skin flap. METHODS: A random pattern skin flap (7 cm x 2 cm) was cranially designed and elevated on the back of a Wistar rat. An image analysis technology was used to evaluate the survival rate of the skin flap, while a biochemistry method was used to test the concentrations of the NO in the tissue. RESULTS: The survival area of the flap in the L-arginine-treated group was significantly enlarged (63.83 +/- 5.13)% (P < 0.01) in seven days postoperatively, compared with the control group (43.26 +/- 2.86)%. The NO concentration in the tissue was no statistic difference between all of the groups immediately after the operation (P > 0.05). But, the NO concentration in the control was decreasing at the beginning and then increasing slightly to reach the high level in 12 hours after the operation. It was thereafter slumped down to the baseline in 72 hours after the surgery. Although the changes in the L-arginine-treated group were quite similar to the control excepting of the extent, the NO concentration was kept in a higher level in the sequential time after the operation (P < 0.01, P < 0.01, P < 0.05 and P < 0.01). CONCLUSIONS: The NO concentration in skin flap tissue after the elevation was going up slightly for a short time. The exogenous L-arginine could promote the NO concentration in the random pattern skin flap to protect it from ischemic injury. PMID- 15131864 TI - [Le Fort III osteotomy and medium position distraction for mid-facial hypoplasia]. AB - OBJECTIVE: To probe the possibility of distraction osteogenesis for correction of mid-facial hypplasia using a new technique. METHODS: The distraction system of mid-facial skeleton consisted of a face-bow, the elastic loops and a device that hitched to bone holes made in the aperture rim. Flllowing Le Fort III osteotomy, a bone hole was drilled at each side of the external-inferior rim of the aperture by a dental bur. The traction device was hitched to the holes through the nostrils. Distraction began three days postoperatively, with the force adjusted dependently upon the rate of progress. When the skeleton reached to the planed position, distraction was retained with a minor force for 8 weeks. Three adults with mid-facial hypoplasia underwent this treatment. RESULTS: The mid-facial skeleton showed a balanced advancement. The patients' facial contour and occlusal relationship recovered completely. CONCLUSIONS: Patients with severe mid-facial hypoplasia could be corrected ideally by the new technique, with balanced advancement of mid-facial skeleton, minor trauma. The design of distraction system was reasonable. The manipulation of the procedure was easy. PMID- 15131865 TI - [Mandibular contour sculpture by osteotomy]. AB - OBJECTIVE: To investigate the method for mandible osteotomy in order to make the mandible of various square face appear harmony. METHODS: According to different types of the mandible, mandible angle osteotomy was performed in combination with mandible edge, mandible half ring osteotomy or chin augmentation. RESULTS: A total of 312 cases have been treated since 1996. In this series, mandible angle and mandible edge osteotomy was performed in 200 cases; only mandible edge osteotomy in 23; mandible half ring osteotomy in 15, chin sharpening in 9, chin augmentation with autogenous bone implantation, in 32. Postoperative follow-up of 150 cases for 1-12 months showed that the satisfactory rate was 97%. CONCLUSION: Integrated mandible osteotomy can make the square face look natural and nice looking. PMID- 15131866 TI - [Effects of NGF and estrogens on human hair follicle in vitro]. AB - OBJECTIVE: To observe the effects of NGF, estrogens and minoxidil on the growth of human hair follicle in vitro. METHODS: In a model of human hair follicle in vitro, the follicle was separately treated with the NGF, estrogens and minoxidil. The growth of the hair follicle was measured in length with an eyepiece micrometer. The effects of the NGF, estrogens and minoxidil were evaluated by measuring the rates of incorporation of 3H-TdR of DNA synthesis. RESULTS: The growth of the human hair follicle was showing significantly faster in the 100 ng/ml NGF and 125 micrograms/ml minoxidil groups, compared with the control (P < 0.05), but the growth was significantly inhibited in the 0.5 microgram/ml 17 beta E2 group (P < 0.05). There was no difference shown for the growth of the hair follicle in the group mixed with 100 ng/ml NGF and 0.5 microgram/ml 17 beta-E2 (P > 0.05). The rates of incorporation of 3H-TdR in the groups were shown that the results just correlated with the results of the above-mentioned method. CONCLUSIONS: The 100 ng/ml NGF and 125 micrograms/ml minoxidil could increase the growth of human hair follicle while the 0.5 microgram/ml 17 beta-E2 could inhibit it. The 100 ng/ml NGF could neutralized the effect of the 0.5 microgram/ml 17 beta-E2. PMID- 15131867 TI - [Collagen changes of allografting an acellular dermal matrix]. AB - OBJECTIVE: To evaluate the collagen changes of allografting an acellular dermal matrix (allo-ADM). METHODS: The allo-ADM was grafted underneath the skin of SD rats. The content of collagen and the proportion of type I to III collagen was examined with a biochemical assay. RESULT: The content of collagen and the proportion of type I to III collagen in the allo-ADM group showed no significant changes. CONCLUSION: The allo-ADM could keep long time in the body and it may be an ideal material for soft tissue filling. PMID- 15131868 TI - [The effects of chitosan on different-derived fibroblasts]. AB - OBJECTIVE: To investigate the effects of chitosan on the biological activities of the fibroblasts derived from different tissues. METHODS: The biological activities of the fibroblasts derived from different tissues were evaluated with a MTT method for fibroblast proliferation, photic and electronic microscope for morphologic and subcellular structure, 3H-proline uptake method for collagen secretion and ELISA box for the secretion of TGF-beta 1, FGF-AB, and IL-8. RESULTS: This study showed that the chitosan inhabited the proliferation of the fibroblasts and the secretion of the TGF-beta 1, FGF-AB and collagen of the fibroblasts with a dose-depended manner in the normal skin, hypertrophic scar and keloid groups, but it stimulated the IL-8. However, there were no significant differences among the three groups (P > 0.05). CONCLUSION: The chitosan could inhibit the growth, proliferation, biosynthesis and secretion of the fibroblasts, and it may be used to treat different scars. PMID- 15131869 TI - [The dynamic expression of TNF-alpha mRNA of hypertrophic scars and its roles]. AB - OBJECTIVE: To investigate the role of tumor necrosis factor alpha: TNF-alpha in the hypertrophic scar and find a valid way to treat the hypertrophic scars with gene therapy. METHODS: TNF-alpha mRNA expression was tested with a semiquantitative reverse polymerase chain reation (RT-PCR) method in the different period of a hypertrophic scar. RESULTS: There was a significant increase of TNF-alpha mRNA as the hypertrophic scar was getting mature (P < 0.01). However, the ratios of TNF-alpha mRNA was significantly in a lower level than the normal scar. CONCLUSIONS: The results suggest that TNF-alpha may play an important role in normal would healing, but the hypertrophic scar may result from the shortage of the TNF-alpha. This indicates that to increase the TNF-alpha with gene therapy may be a good way to treat the hyphertrophic scar. PMID- 15131871 TI - Valuing your office staff goes a long way. PMID- 15131870 TI - [Effect of lanthanum chloride on expressions of collagen protein in a wound tissue]. AB - OBJECTIVE: To evaluate the effect of lanthanum chloride on expressions of collagen protein and find a way to prevent and treat scar. METHODS: Four linear incisions were made on the dorsal skin of an adult, female Sprague-Dawley rat as an animal model. One was non-manipulated as a control; the second was injected with distilled water as a sham-control; the third was injected with 50 mmol/L of lanthanum chloride, and the fourth was injected with 50 micrograms neutralizing antibody of TGF-beta 1 as a positive control. All of the wound tissues were harvested and assayed with ABC method in 14 days and 28 days after the surgery. RESULTS: The expressions of type I, III and IV of collagen protein in the third group significantly reduced in 14 and 28 days after the operation, compared with the control or sham-control group. Its values wen as similar as the fourth group. CONCLUSION: Lanthanum chloride could inhibit the expressions of collagen protein, and it may be used to prevent and treat scars. PMID- 15131872 TI - MDS member survey on auxiliary employment and retention and dental hygiene focus groups: results and recommendations. PMID- 15131873 TI - 10 strategies for successful staff retention. PMID- 15131875 TI - Using auxiliaries--legally, efficiently, and effectively. PMID- 15131874 TI - Facing the challenge: recruitment & retention of staff. AB - The economic slowdown has begun to ease the shortage of clinical staff. Programs in the accredited dental assisting and dental hygiene programs are fully subscribed. As women and men alike look for alternative careers, they are looking at dental hygiene and dental assisting for the first time. It is our responsibility as dental professionals to spread the word--dental careers are rewarding and fulfilling. Using an effective hiring strategy, we will be able to compete with the corporate world and recruit and retain competent staff. PMID- 15131876 TI - Dentistry and the law. Avoiding employee lawsuits: 10 best practices for dental offices. PMID- 15131877 TI - Interviewing job applicants for a staff opening. PMID- 15131878 TI - An interview with Susan J. Deliddo, RDH. PMID- 15131879 TI - Is insurance important to the dental practice? PMID- 15131880 TI - The path to long-term retention. PMID- 15131882 TI - How to properly analyze a resume. PMID- 15131881 TI - 10 ways to find your ideal job candidate. PMID- 15131883 TI - Enlist auxiliaries as part of the purchasing process. PMID- 15131884 TI - Employing quality staff--your best investment. AB - For the student graduating this spring or for the dentist in practice a year or two, my message is the same. The best investment you can make in your future is to hire and keep the highest quality staff. A dentist new to the role of employer may have neither received the human resources training of the needed skills to attract, interview, and hire quality staff nor received legal advice on how to perform as an employer who is legally in compliance with federal and state laws. PMID- 15131885 TI - Assemble a WOW team. PMID- 15131886 TI - Facility profile. Desert clinic blends beauty, efficiency. PMID- 15131887 TI - Tooling around. Managers consider reliability-centered maintenance techniques. PMID- 15131888 TI - Worlds collide. Clinical engineering is converging with information technology. Are you ready? PMID- 15131889 TI - Using landscaping & exterior design elements to enhance campus wayfinding. Guiding sites. PMID- 15131890 TI - Book it! An overview of ASHRAE's design manual for hospitals and clinics. PMID- 15131891 TI - Tough love. Though designed for heavy-duty use, coated floors still require care. PMID- 15131892 TI - Two-color immunostaining of liver fine needle aspiration biopsies with CD34 and carcinoembryonic antigen. Potential utilization in the diagnosis of primary hepatocellular carcinoma vs. metastatic tumor. AB - OBJECTIVE: To examine immunohistochemical staining of cell block material with antibodies against vascular marker CD34 and polyclonal carcinoembryonic antigen (pCEA) for their clinical utility as part of a 2-color staining protocol in fine needle aspiration (FNA) biopsy of liver masses to distinguish metastases from primary hepatocellular carcinoma (HCC). STUDY DESIGN: The authors obtained cell block material from 96 liver FNAs and performed simultaneous (i.e., "dual-color") immunohistochemical staining utilizing antibodies against vascular marker CD34 and pCEA. Cases were blinded and evaluated by the authors for staining pattern and intensity. A consensus was obtained, the results were unblinded, and the diagnoses were correlated. RESULTS: After staining, 89 cases had sufficient tissue for evaluation. Of the 19 HCC cases, 16 (84%) showed peripheral staining with CD34, and 13 (68%) showed a canalicular or mixed canalicular-cytoplasmic staining pattern for pCEA. Thirteen cases (68%) showed staining for both antigens. All HCC exhibited immunostaining for at least 1 antibody in an appropriate staining pattern. Of the 67 cases of metastatic malignancy, 5 (7%) showed a predominantly transgressing pattern of CD34 staining, 43 (64%) showed a predominantly cytoplasmic or mixed cytoplasmic-canalicular pattern of pCEA staining, and 2 cases (3%) showed staining for both antigens in a transgressing CD34 pattern and cytoplasmic pCEA pattern. None of the 3 normal liver tissue blocks showed staining with either antigen. CONCLUSION: Two-color immunohistochemical staining of liver cell block material obtained by FNA with antibodies to CD34 and pCEA can be helpful in differentiating metastatic tumors vs. primary HCC. PMID- 15131893 TI - Regional heterogeneity of EGFR gene amplification and nuclear morphology in glioblastomas. An investigation using laser microdissection and pressure catapulting. AB - OBJECTIVE: To study the regional heterogeneity of epidermal growth factor receptor (EGFR) gene amplification (EGFR-GA) in glioblastomas, considering the relationship between this mutation and morphology of tumor cell nuclei. STUDY DESIGN: Tissue samples gained by laser microdissection and pressure catapulting were used for the performance of differential polymerase chain reaction in 32 morphologically different regions from 7 glioblastomas. Semiquantitative determination of EGFR expression and image analysis of tumor cell nuclei were performed in the same regions. RESULTS: Distinct regional differences concerning the degree of EGFR-GA were found in 2 tumor cases. When comparing regions with different degrees of gene amplification within these cases, morphologic differences in tumor cell nuclei were observed. The other tumor cases also showed distinct intratumoral heterogeneity concerning histomorphology but no regional heterogeneity in the degree of EGFR-GA. When comparing regions with a low densitometric EGFR/interferon (INF) band ratio (< 2.19, n = 18) and a high EGFR/IFN band ratio (> 4.39, n = 14), the latter type of region showed a significantly higher percentage of Ki-67--positive tumor cell nuclei and lower values for several shape variables (Fourier amplitudes), indicating a tendency toward a more regular nuclear shape in regions with distinct EGFR-GA. For the EGFR/IFN band ratio, a significant correlation was found with several morphometric variables, especially those of nuclear shape and distances between nuclei. CONCLUSION: In glioblastomas showing regional heterogeneity in the degree of EGFR-GA, morphology of tumor cell nuclei has been shown to be different when comparing regions with different degrees of EGFR-GA. Glioblastomas may also show distinct regional heterogeneity of histomorphology without evidence of regional heterogeneity of EGFR-GA. A significant statistical association has been confirmed between the degree of EGFR-GA and quantitative morphology of tumor cell nuclei. PMID- 15131894 TI - Computer-based malignancy grading of astrocytomas employing a support vector machine classifier, the WHO grading system and the regular hematoxylin-eosin diagnostic staining procedure. AB - OBJECTIVE: To investigate and develop an automated technique for astrocytoma malignancy grading compatible with the clinical routine. STUDY DESIGN: One hundred forty biopsies of astrocytomas were collected from 2 hospitals. The degree of tumor malignancy was defined as low or high according to the World Health Organization grading system. From each biopsy, images were digitized and segmented to isolate nuclei from background tissue. Morphologic and textural nuclear features were quantified to encode tumor malignancy. Each case was represented by a 40-dimensional feature vector. An exhaustive search procedure in feature space was utilized to determine the best feature combination that resulted in the smallest classification error. Low and high grade tumors were discriminated using support vector machines (SVMs). To evaluate the system performance, all available data were split randomly into training and test sets. RESULTS: The best vector combination consisted of 3 textural and 2 morphologic features. Low and high grade cases were discriminated with an accuracy of 90.7% and 88.9%, respectively, using an SVM classifier with polynomial kernel of degree 2. CONCLUSION: The proposed methodology was based on standards that are common in daily clinical practice and might be used in parallel with conventional grading as a second-opinion tool to reduce subjectivity in the classification of astrocytomas. PMID- 15131895 TI - Mean nuclear volume in squamous cell carcinoma of the vulva. AB - OBJECTIVE: To compare mean nuclear volume (MNV) estimated by the stereologic intercept method in lymph node-positive and -negative cases of squamous cell carcinoma of the vulva. STUDY DESIGN: This retrospective study consisted of 53 cases (lymph node metastasis, n = 19; cases without lymph node metastasis, n = 34) of squamous cell carcinoma of the vulva. MNV was estimated with the help of an image cytometer. The nuclear point intersection method was used to measure MNV. The mean nuclear volumes of both lymph node-positive and -negative cases were compared. RESULT: MNV in the lymph node-negative and -positive cases was 717.0 +/- 533.1 and 1,961.4 +/- 1,369.6 microns 3, respectively (P < .000, Mann Whitney U test). There was a significant difference in MNV between the 2 groups of tumors. CONCLUSION: The observations from the present study suggest that estimation of MNV of malignant squamous cells from the vulva on conventional histopathology sections may provide an objective and useful diagnostic tool in predicting lymph node metastasis. PMID- 15131896 TI - Karyometric features of low and high grade glial tumors as compared with their MRI appearance. AB - OBJECTIVE: To compare the results of a magnetic resonance imaging (MRI) grading designed to identify low and high grade gliomas with karyometry used as a tool to grade primary brain tumors. STUDY DESIGN: A consecutive series of 23 primary brain tumors was selected for this study. The neuroradiologist, not knowing the histologic diagnoses, divided the cases into low and high grade categories on the basis of the following 7 features: border sharpness, heterogeneity without contrast, cavitation, contrast enhancement, hypervascularity, mass effect and perifocal T2 hyperintensity. To each feature was given a numerical value, ranging from 1 to 3. All the cases were reviewed and classified by the same pathologist, blinded to the MRI diagnosis. Two hundred nuclei per case were recorded, and 93 karyometric features related to nuclear area, total optical density and chromatin distribution were analyzed for each nucleus. Statistical analysis included discriminant analysis, Kruskal-Wallis test, nonsupervised learning algorithm P index and Beale statistic. RESULTS: Ten cases were classified as low grade on the basis of their MRI features. The corresponding histopathologic diagnoses were: grade 2 astrocytoma in 2 cases and grade 2 oligodendroglioma in 8 cases. An MRI diagnosis of high grade tumor was made in 13 cases. In 10 cases it was confirmed by the histopathologic diagnosis (3 grade 3 astrocytomas, 1 grade 3 oligodendroglioma and 6 glioblastomas). In the remaining 3 cases the histologic examination revealed a low grade tumor, 1 grade 2 astrocytoma and 2 grade 2 oligodendrogliomas. For the purposes of the karyometric analysis the cases were allocated to the low or high grade category according to their histologic diagnosis (13 cases low grade and 10 cases high grade). Nuclei from low and high grade tumors showed clearly different karyometric characteristics. The oligodendroglioma nuclei had abnormality values close to the low grade standard, while the astrocytoma nuclei were a highly dispersed group with characteristics indicative of a higher degree of nuclear abnormality than the oligodendroglioma nuclei. The results of karyometric analysis showed that grade 2 tumors, corresponding to the low grade group, form a rather distinct category from grade 3 and 4 tumors belonging to the high grade group. CONCLUSION: The results of MRI grading based on a series of features that are routinely assessed by the neuroradiologist to reach a final diagnosis correlate highly with the histopathologic diagnosis. Karyometry can be a useful adjunct to histologic grading. PMID- 15131897 TI - Choice of methodology for quantifying cancer structures in tissue sections. A comparison of 2- and 3-dimensional estimators of mitotic activity, cellularity and nuclear size in breast cancer. AB - OBJECTIVE: To analyze the relationships between a range of 2-(2D) and 3 dimensional (3D) estimators of important tumor features (mitotic activity, cellularity and nuclear size). STUDY DESIGN: Measurements were performed in systematically sampled fields of vision of 3-microns-thick sections and in optical disectors of 40-microns-thick sections of 93 breast cancers. RESULTS: 2D mitotic profile density and frequency correlate highly with 3D mitotic density and frequency (r > or = .75); the choice of estimator was of minor importance. 2D nuclear profile density correlated quite closely with 3D nuclear volume fraction and nuclear density (r > or = .66). Cellularity estimators were, however, influenced differently by nuclear size. 2D mean nuclear profile area and 3D volume- and number-weighted mean nuclear size correlated less closely (r > or = .51). Tumors with high mitotic counts generally had large nuclei, reflecting the fact that both variables are associated with aggressiveness. CONCLUSION: 2D and 3D quantitative estimates of corresponding tumor features often correlate closely. Easily applicable and unbiased 3D techniques are recommended for obtaining the mean size of nuclei (or other particles) and of volume fractions of structures. For ranking of cellularity and mitotic activity, 2D measurements are usually sufficiently accurate and close to corresponding 3D estimates. However, if exact quantities of tissue structures are required, unbiased (3D) techniques must be used. PMID- 15131898 TI - Correlation between VEGF expression and angiogenesis in breast carcinoma. AB - OBJECTIVE: To analyze the role of vascular endothelial growth factor (VEGF) secreted by tumor cells in angiogenesis of breast carcinoma using image morphometry. STUDY DESIGN: Thirty-four cases of node-negative breast carcinoma were used in the study. There were 6 grade 1, 20 grade 2 and 8 grade 3 tumors. For each case, 2 consecutive sections from the same block were cut. Immunostaining for VEGF and CD31 was carried out, and areas of highest staining density were marked. Those marked "hot spots" for CD31 and VEGF for each case were subsequently compared morphometrically. The area and intensity of immunostaining on each slide were also scored. RESULTS: The total scores for VEGF and CD31 were 5.15 and 3.79, respectively. All 34 cases showed cytoplasmic positivity for VEGF within the tumor cells. The average number of hot spots for VEGF and CD31 were 2.41 and 2.47, respectively, and the average number of hot spots that matched between these 2 groups were 0.79. Statistical analysis using Pearson's coefficient of correlation showed no significant match between the hot spots for CD31 and VEGF. Also, there was no significant difference between the total scores of CD31 and VEGF. CONCLUSION: VEGF is expressed in most breast carcinomas. However, the lack of topographic correlation between microvessel density and VEGF expression supports the notion that multiple angiogenic factors may play a role along with VEGF in the angiogenic process. PMID- 15131899 TI - Fractal dimension in aspiration cytology smears of breast and cervical lesions. AB - OBJECTIVE: To standardize the automated measurement of fractal dimension on cytologic smears and compare the fractal dimension of benign and malignant breast cells and cervical lesions on cytologic material to evaluate its role in the discrimination of benign from malignant cells. STUDY DESIGN: We randomly selected fine needle aspiration cytology smears of 42 cases of infiltrating duct carcinoma and 38 cases of fibroadenoma of the breast. Similarly, 16 cervical carcinoma and 20 normal cervical smears were selected for study. Ten cells were selected randomly from each case. Box counting of fractal dimension of malignant and benign cells was achieved with an image cytometer (Leica, Cambridge, England) using Quantimet 600 software (Leica). Then a well-spaced grid with multiple small boxes of a particular pixel length was superimposed on the cell. The dimension of the box was selected as 4, 8 and 16 pixels. With the help of a logical "AND" operation, we counted the number of boxes touching the peripheral margin of the cell nuclei. For each cell, the log-log graph of 1 per box size was plotted against the number of boxes touching the peripheral rim of the cell. The slope of each graph was identified using the least-squares method of regression analysis. RESULTS: The mean fractal dimension of malignant cells was 0.8536 +/- 0.1120 as compared to 0.8403 +/- 0.1115 in benign cell groups. The Mann-Whitney U test showed a significant difference in fractal dimension in these 2 groups (P = .05). The mean fractal dimension of malignant cells from the cervix was 0.8656 +/- 0.1499 as compared to 0.8315 +/- 0.1312 in benign cells. The Mann-Whitney U test showed a significant difference in fractal dimension in these 2 groups (P < .02). CONCLUSION: Fractal dimension may be a helpful adjunctive technique to discriminate between benign and malignant cells. PMID- 15131900 TI - Fractal dimension in endometrial carcinoma. AB - OBJECTIVE: To mathematically assess in a pilot study, endometrial glandular margin irregularity in simple hyperplasia, complex atypical hyperplasia and well differentiated endometrial carcinoma with the help of box counting of fractal dimension and to discriminate these lesions on the basis of box counting of fractal dimension of the gland. STUDY DESIGN: Ten cases each of endometrial simple hyperplasia (without atypia), complex hyperplasia with atypia and endometrial carcinoma (well-differentiated, endometrioid) were assessed in the study. Five fields at 20 x magnification from each case were randomly selected, and the glands were outlined with the help of a pointer. Using the box counting method, the fractal dimension of each case was measured. RESULTS: Mean fractal dimension in simple hyperplasia, complex atypical hyperplasia and endometrial carcinoma was, 0.899 +/- 0.13, 0.932 +/- 0.042 and 0.939 +/- 0.02, respectively. Statistical analysis showed that the fractal dimension of glands of simple hyperplasia were significantly different from that of complex atypical hyperplasia and endometrial carcinoma (P = .041 and .013, respectively, ANOVA). However, there was no significant difference in fractal dimension between glands of complex hyperplasia and of endometrial carcinoma (P = .659, ANOVA). CONCLUSION: This study provides mathematical (objective) assessment of the measurement of glandular margin irregularities in simple hyperplasia, complex atypical hyperplasia and endometrial carcinoma. Fractal dimension of gland margin may have diagnostic potential in the future. PMID- 15131901 TI - Morphometric image analysis of follicular lesions of the thyroid. AB - OBJECTIVE: To determine the role of image morphometry in distinguishing various follicular lesions of the thyroid in cytologic smears. STUDY DESIGN: Archival fine needle aspiration smears of 10 cases each of follicular hyperplasia, follicular adenoma, follicular carcinoma and follicular variant of papillary carcinoma were used for the study. All cases were histopathologically proven. At least 100 random nuclei from each case were subjected to analysis with an image cytometer. Area, convex area, length, width, perimeter, convex perimeter and roundness of nuclei were measured using a 40 x objective (1 pixel = 0.446 micron). RESULTS: ANOVA showed that all the nuclear variables studied were significantly different (P < .05) in follicular hyperplasia as compared to follicular carcinoma and papillary carcinoma. All nuclear variables except roundness were also significantly different (P < .05) between follicular hyperplasia and follicular adenoma. However, between follicular adenoma, follicular carcinoma and papillary carcinoma there was considerable overlap of nuclear morphometric parameters. CONCLUSION: Image morphometry may help to distinguish nonneoplastic follicular lesions (hyperplasia) from neoplastic lesions (adenomas and carcinomas). However, to distinguish benign from malignant follicular lesions, image morphometry might not improve the accuracy of standard cytologic examination. PMID- 15131902 TI - The four-part patient. PMID- 15131904 TI - 10 minutes with Bill Brown. Interview by Raphael M. Barishansky. PMID- 15131903 TI - Situational perceptiveness. Assess the predicament--not just the physiology! PMID- 15131905 TI - EMS response time standards. PMID- 15131906 TI - Bioterrorism awareness for EMS. AB - It is important to understand that the issues surrounding bioterrorism and all weapons of mass destruction are complex. In an effort to enhance response to such events, EMS should handle all incidents from the perspective of an all-hazards approach. Prevention, preparation, response and recovery are essential to the safe mitigation of all incidents. Organizations must be prepared. Plan now for a safer tomorrow. Your personnel and communities depend on you. PMID- 15131907 TI - Pathophysiology of asthma. PMID- 15131908 TI - Botulinum toxin: a bioterrorism weapon. PMID- 15131909 TI - Explosive events. EMS response to a bombing incident. PMID- 15131910 TI - Hazardous-materials antidotes. PMID- 15131911 TI - Chronic pain: the role of EMS. PMID- 15131912 TI - Vasopressin: improving resuscitation? AB - Historically, epinephrine has been used in the treatment of cardiac arrest for its alpha effects of peripheral vasoconstriction to promote vital organ perfusion. Unfortunately, epinephrine, like many other pharmacological agents, targets other receptor sites that can have a detrimental effect on the patient in cardiac arrest. Vasopressin is an endogenous hormone that may be an ideal alternative or adjunctive to epinephrine in the setting of cardiac arrest. Vasopressin, like epinephrine, promotes selective but potent vasoconstriction of smooth muscle, but unlike epinephrine, without the potentially harmful side effects of increasing myocardial workload, therefore increasing oxygen demand and subsequent worsening of cardiac function. The newest data on the impact of vasopressin in cardiac arrest is promising. Further studies are required to determine if vasopressin has a significantly positive impact in outcome as measured by discharge of the neurologically intact patient in the North American EMS model. PMID- 15131914 TI - Secrets revealed for male catheterization. PMID- 15131913 TI - Professional etiquette. How you show your respect for people. PMID- 15131915 TI - Losing it all. PMID- 15131916 TI - Quality of life among men treated with brachytherapy for prostate cancer. AB - Brachytherapy, the implantation of permanent radiation sources into the transperineal area, is being used with increasing frequency as treatment for localized prostate cancer. In this study, men undergoing brachytherapy evaluated their quality of life (QoL) before and after treatment. Brachytherapy affects QoL within the domains of emotional well-being and specific prostate concerns. Knowledge of the treatment effects on QoL will assist nursing staff working with patients faced with treatment decisions for prostate cancer. PMID- 15131917 TI - Getting ready for certification: voiding dysfunction. PMID- 15131918 TI - The impact of self-care practices on treatment of interstitial cystitis. AB - Self-care, as a theoretical base of practice in caring for patients with interstitial cystitis (IC), has been suggested as a model of holistic health. Those living with IC look to the nurse/health care practitioner for intervention and care. Nurses operating from the self-care nursing framework may find this model useful in caring for patients with IC. These patients are educated by the nurse regarding preventative measures, interventions, and advanced treatments and therefore may be in a better position to participate in achieving the goal of optimal health. Self-care nursing helps the client to care for themselves through education, resource acquisition, and role-modeling positive behavioral outcomes. PMID- 15131919 TI - Interstitial cystitis and self-care: bearing the burden. AB - The purpose of this case study article is to place a personal twist on a frightening disease process known as interstitial cystitis (IC). As nurses, the goal of our interventions is to promote the independence of the client in his/her own self-care. One nursing theory, Modeling and Role-Modeling (Erickson, Tomlin, & Swain, 1983), has assisted me not only as a nurse but also as a client to overcome the IC dilemma through the practice of self-care nursing. PMID- 15131920 TI - Fad diets and obesity--Part I: Measuring weight in a clinical setting. AB - Obesity is a recognized epidemic in many regions around the world and billions of dollars are spent each year in attempting to combat this problem. However, before a discussion of the different conventional and alternative treatments for obesity can be initiated, it is first critical to determine whether or not a certain individual is actually overweight, obese, or has an excess of adipose tissue. Therefore, a review of the various popular and unpopular measurements of obesity is needed. A variety of measurements exist such as bioelectrical impedance, body mass index (BMI), crude weight, densitometry, dual energy x-ray absorptiometry (DEXA), lean body mass (LBM), skinfold thickness, and waist-to-hip ratio (WHR). All of these measurements contain inherent advantages and disadvantages, but many of these can still be used in a clinical setting. Health professionals should acquaint themselves with these different measurements in order to take the first step in bringing attention to and potentially treating a condition that affects virtually every medical discipline. PMID- 15131921 TI - Practical cystoscopy: care of the instruments and preparation for a cystoscopic examination. AB - Periodically, a look at the past helps health care professionals see how much practice has changed as well as how some concepts and practices remain similar to the original rationales and techniques. Two chapters from a 1911 textbook on cystoscopy have been reprinted that describe catheter and instrument care and preprocedure requirements for cystoscopy performed in the hospital, office, or home. Sterilization methods and instruments for cystoscopy have improved in the century since this book was written but the rationale for the procedure and concepts guiding the instrumentation have some similarity with the practices performed nearly 100 years ago. Some comments also provide a bit of comic relief. One example, "this undoubtedly shortens the life of the catheter, but it lengthens the life of the patient" appears in the discussion about catheter cleaning after use in a pus case. PMID- 15131922 TI - Update on SARS. PMID- 15131923 TI - Procedures, ethics, and funding sources. PMID- 15131924 TI - Warming your heart: the energy solution. AB - Citron (2002) probably sums it up best when he states that generosity is the best strategy for corporate prosperity. He believes that the love you get from others as a person and a corporation is equal to the love you extend to others. It's really about creating communities of caring and missions of inclusiveness. And the way to make this all happen is to continually recharge our batteries by staying close to the beautiful stories of our organization that will undeniably warm our hearts and give us the energy to be generous. PMID- 15131925 TI - Evidence-based radiology problems. Radiofrequency ablation of lung malignancy April 2004-March 2005. PMID- 15131926 TI - An alternative clip-marking method for use after 14-gauge large core needle biopsy of the breast. PMID- 15131927 TI - Multidetector computed tomographic assessment of coronary ostial pseudoaneurysms: case report. PMID- 15131928 TI - Preparing an educational exhibit. PMID- 15131929 TI - Limited-sequence magnetic resonance imaging in the evaluation of the ultrasonographically indeterminate pelvic mass. AB - OBJECTIVE: To evaluate the usefulness of limited-sequence magnetic resonance imaging (MRI) in the elucidation of ultrasonographically indeterminate pelvic masses. This study focused only on pelvic masses in which the origin of the mass (uterine v. extrauterine) could not be determined by ultrasonography (US). The origin of a pelvic mass has clinical implications. A mass arising from the uterus is most likely to be a leiomyoma, which is a benign lesion, whereas an extrauterine mass will have a higher likelihood of malignancy and usually requires surgery. METHODS: Eighty-one female patients whose pelvic mass was of indeterminate origin on US also underwent limited-sequence MRI of the pelvis. Most of the MRI examinations were performed on the same day as the US. Limited sequence MRI sequences included a quick gradient-echo T1-weighted localizer and a fast spin-echo T2-weighted sequence. Final diagnoses were established by surgical pathology or by clinical and imaging follow-up. RESULTS: Limited-sequence MRI was helpful in 79 of the 81 cases (98%). Fifty-two of the 81 masses (64%) were leiomyomas. One was a leiomyosarcoma. The extrauterine masses (26/81 [32%]) were identified as 14 ovarian malignancies, 4 endometriomas, 3 dermoids, an ovarian fibroma, an infarcted fibrothecoma, an infarcted hemorrhagic cyst, a sigmoid diverticular abscess and a gastrointestinal stromal tumour of the ileum. In the other 2 cases (2/81 [2%]), the origin of the pelvic mass remained indeterminate. Both of these indeterminate masses showed low signal on T2-weighted images and were interpreted as probable leiomyomas. They were not surgically removed but were followed clinically and had a stable course. CONCLUSION: Limited-sequence MRI is a quick and efficient way to further evaluate ultrasonographically indeterminate pelvic masses. Limited-sequence MRI of the pelvis can suffice, in these cases, without requiring a full MRI examination. PMID- 15131930 TI - Magnetic resonance imaging of the heart and great vessels. PMID- 15131931 TI - The spectrum of radiologic findings in idiopathic arterial calcification of infancy: pictorial essay. PMID- 15131932 TI - Balloon angioplasty versus primary stenting of ostial atherosclerotic renal arterial stenoses: critically appraised topic. PMID- 15131933 TI - Evaluation of carotid artery stenosis: contrast-enhanced magnetic resonance angiography compared with conventional digital subtraction angiography. AB - OBJECTIVE: To compare retrospectively high-resolution contrast-enhanced magnetic resonance angiography with centric k-space filling and digital subtraction angiography in the assessment of degree of carotid artery stenosis. METHODS: Two neuroradiologists independently evaluated 14 carotid arteries (in 7 patients), and the degree of carotid stenosis determined was compared to calculate interobserver reliability. Thirty-six carotid arteries (in 18 patients) were then assessed with both contrast-enhanced magnetic resonance angiography and digital subtraction angiography. The degree of stenosis was determined by consensus, and the findings of the 2 modalities were compared. RESULTS: Close interobserver agreement was found with an intraclass correlation coefficient of 0.98 (95% confidence interval 0.95-0.99). Sensitivity of 86% and specificity of 91% were found for contrast-enhanced magnetic resonance angiography detection of surgically significant carotid stenosis compared with digital subtraction angiography. The Spearman rank correlation test also found a significant correlation (R(s) = 0.90, p < 0.001) in the comparison of the classifications of degree of carotid artery stenosis. CONCLUSION: Contrast-enhanced magnetic resonance angiography shows excellent correlation with digital subtraction angiography for the evaluation of carotid artery disease and has the potential to replace it in the assessment of degree of carotid artery stenosis. PMID- 15131934 TI - Residents' corner. Answer to case of the month #95. Endometrioma of the bladder. Endometrioma of the bladder. PMID- 15131935 TI - Residents' corner. Answer to case of the month #96. Right frontal lobe ganglioglioma. PMID- 15131936 TI - The dire cost of obesity. PMID- 15131937 TI - Keys to passing the JCAHO inspection. PMID- 15131938 TI - Practice, practice. Overcoming barriers to evidence-based nursing. PMID- 15131939 TI - Change is in the air. Options to ensure worker safety. PMID- 15131940 TI - Stats. Getting a lift. Costs of cosmetic surgery supplies on the rise. PMID- 15131941 TI - Governor Taft signs prescriptive authority bill. PMID- 15131945 TI - Terrorism to action: a year later. PMID- 15131946 TI - Position on immigration and the nursing workforce. PMID- 15131947 TI - What kinds of malpractice cases are happening around the state? PMID- 15131948 TI - Race against time. PMID- 15131949 TI - Hearts and minds. PMID- 15131950 TI - Recovered lives. PMID- 15131951 TI - Talking trees. PMID- 15131952 TI - Talking cure. PMID- 15131953 TI - Welfare writes. PMID- 15131954 TI - Lone voice. PMID- 15131955 TI - Working with difference. PMID- 15131956 TI - Unlocking our potential. PMID- 15131957 TI - What will people think? PMID- 15131958 TI - Engaging communities. PMID- 15131959 TI - Loose nuts. PMID- 15131960 TI - Cassandra. PMID- 15131961 TI - This life. PMID- 15131962 TI - [Inheritance of heading date in crosses of wheat cultivar responsiveness to light intensity]. AB - The inheritance of heading date has been studied in crosses of spring bread wheat cultivars differing in their reaction to various light intensities. The parental cultivars were similar in their Vrn and Ppd gene systems and differed in heading dates. Domination of higher reaction to light intensity has been shown in F1 on duration of "ear emergence-heading" period. Analysis of F2 populations has shown the identity of earliness per se genetic systems in Novosibirskaya 22 and Ordynskaya cultivars. Digenic differences have been revealed for both these cultivars from Tyumenskaya rannyaya cultivar on duration of "ear emergence- heading" period. PMID- 15131963 TI - [Comparative study of allozyme polymorphism in groups of pine trees (Pinus sylvestris L.) with different seed productivity]. AB - Genotypes of 196 Pinus sylvestris L. plants from 10 natural populations of five Ukrainian regions have been determined using 19 polymorphic isozyme loci. Variability of quantity of full-grained, empty-grained and underdeveloped seeds in the cones of these plants has been studied. The basic indexes of genetic polymorphism were determined for 6 samples presented by 18-19 trees with high and low productivity of the full-grained, empty-grained and underdeveloped seeds. The maximum amount of rare alleles and genotypes as well as the highest heterozygosity (Ho = 0.285) were typical for the sample of plants with the maximum quantity of empty-grained seeds in the cones. PMID- 15131964 TI - [Identification of sunflower genotypes using microsatellite markers]. AB - Polymorphism of some DNA microsatellite sequences of sunflower breeding genotypes (inbred lines, hybrids) was investigated. Allelic composition on 8 SSR loci is unique characteristic of every analysed genotype allowing their identification. Microsatellite markers are proposed to be used for definition of line genetic purity and of F1 seeds hybridity range. PMID- 15131965 TI - [Effect of culture medium composition on callus formation and regeneration in oil flax anther culture]. AB - Regeneration ability in callus cultures from anthers of two hybrid genotypes of oil flax was studied on N6 and LMA-1 nutrient media at various concentrations of cytokinine 6-benzylamynopurine (BAP). It was shown that callus grew and developed better at BAP concentrations of 2 mg/l, comparing with 4 and 6 mg/l. Shoot and root regeneration was observed in F1 genotype 6-8-gnezdny x M22 only and did not depend on BAP concentration in the medium and on the medium composition itself. Transfer onto fresh medium often stimulated dedifferentiation of the regenerated structures. PMID- 15131966 TI - [Cytogenetic investigation of geranium plants in tissue culture]. AB - Cytogenetic study of six cultivars and six selection lines of geranium (Pelargonium spp.), as well as of 100 plants regenerated from callus cultures has been performed. The majority of cultivars and lines had somatic chromosome numbers 2n = 7x = 56. Among regenerated plants of different cultivars (Rozovaja, Dushistaja, Krunk, Aist, Regar) obtained in vitro from various types of explants (internode, petiole) 61% of diploids and 39% aneuploids were revealed. Chromosome numbers in aneuploids varied from 46 to 82, among them 25.6% regenerated plants had 2n = 72; 10.2% -2n = 68; 5.1% -2n = = 64 and 12.8% -2n = 62. Addition of colhicine to nutrient medium increased the number of aneuploid plants. PMID- 15131967 TI - [Genetic variation of carrion and hooded crows and their hybrids based on RAPD PCR data]. AB - RAPD-PCR analysis of genetic variability of carrion crows, hooded crows and their phenotypic hybrids from the zones of overlap of their areals and hybridization of parental forms in Siberia has been carried out. According to RAPD variability the following parameters were estimated for both species and their hybrids: genetical distances (DN), polymorphism (P95), mean expected heterozygosity (H(e)), gene fixation coefficient (Fst) and the portion of interpopulation differentiation (Gst). It was shown that the rate of genetic variability in carrion crows and hooded crows is less than in the hybrids. It turned out that the hybrids were genetically closer to each other (DN = 0.295) than to both parental species (DN = 0.441 and 0.397 with C. corone and C. cornix, correspondingly). The studied individuals of C. cornix show the minimal level of heterozygosity and polymorphism (H(e) = 0.12 and P95 = = 27.8%) in comparison with C. corone and the crows of hybrid zone (H(e) = 0.18 and 0.20; P95 = 42.6% and 50.4%, correspondingly). Right's indexes of inbreeding (Fst = 0.285) and population spliting (Gst = 0.352) indicate low genetic variability in all the analysed birds. Nevertheless UPGMA and NJ dendrogrammes differentiate the individuals from the areals of carrion crows, hooded crows and from hybrid zone to different clusters. Our data correlate with molecular-genetic investigations of Corvidae carried out with other methods, and do not contradict the conception of carrion crows and hooded crows as semispecies in the context of the conception of overspecies. PMID- 15131968 TI - [Antimutagenic activity of plant extracts from Armoracia rusticana, Ficus carica and Zea mays and peroxidase in eukaryotic cells]. AB - Antimutagene activity and high efficiency of antimutagene action of plant extracts from horseradish roots (Armoracia rusticana), fig brunches (Ficus carica) and mays seedlings (Zea mays) and their ability to decrease the frequency of spontaneous and induced by gamma-rays chromosome aberrations in meristematic cells of Vicia faba and marrow cells of mice have been shown. Comparative assessment of genoprotective properties of peroxidase and the studied extracts has revealed higher efficiency of antimutagene action of peroxidase. PMID- 15131969 TI - [Intra-population differentiation of Cholmogoric purebred cattle for the gene frequency of the kappa-casein locus]. AB - Investigation of the sampling of 2907 cows from 17 farms of three regions of the Non-chernozem zone of the European part of Russia showed that kappa-caseine synthesis is controlled by two allelic genes--ae-CnA and ae-CnB. The average frequency of the ae-CnA was 0.689; of the ae-CnB--0.311; in factory herds--0.718 and 0.282, respectively; in reproductive herds--0.714 and 0.286, respectively; in market herds--0.629 and 0.371, respectively. The high frequency of ae-CnB allele (0.331) has been revealed in subpopulation of cholmogoric cattle of the Komi Republic. In the herds of cholmogoric cattle of the flood-plains of Pechora river ae-CnB frequency reached 0.520 and 0.575. The assumption is made about selective role of polymorphism and necessity of rational use of gene pool of domestic cattle. PMID- 15131970 TI - [Cytogenetic features of the differential diagnosis of lymphoid thymomas, small cell non-Hodgkin lymphomas and undifferentiated small cell lung carcinoma]. AB - The predominance of compact and nucleolonemic types of nucleoli in undifferentiated small cell carcinoma of lung, the prevalence of micronucleoli and ring-shaped types of nucleoli in lymphoid thymoma and the increase of the level of micronucleoli in small cell non-Hodgkin's lymphoma have been established. PMID- 15131971 TI - [Premature condensation of sperm chromosomes in human oocytes fertilized in vitro]. AB - Analysis of the frequency of oocytes containing premature condensed sperm chromosomes depending on clinical and embryological parameters has been carried out. It was shown that this frequency comes to 11.6% during in vitro fertilization and depends on morpho-functional characteristics of acnipo[symbol: see text] oocytes. PMID- 15131972 TI - [Comparative analysis of the frequency and type of chromosomal aberrations in somatic cells in maternal-fetal infections]. AB - Fetal infections have high specific gravity in the structure of perinatal diseases and mortality. The gravity of infection processes in pregnancies and degree of fetal pathology are not correlated. Latent asymptomatic forms of pregnant's diseases can often be the cause of the congenital malformations and fetal death. We present here results of the analysis of the frequencies and types of chromosome aberrations in pregnancies with latent infections. The chromosome structure damages, mainly chromatide-type aberrations, were registered with higher frequency in the studied group. PMID- 15131973 TI - [Modification of lipopolysaccharides as one of the ways of adaptation of pathogenic bacteria and their populations to biotic and abiotic environmental factors]. AB - The review is devoted to a problem of adaptive role of lipopolysaccharide (LPS) modification of pathogenic bacteria cells. The biological activity of LPS depends on their molecular conformation, which is determined by primary structure and charges of their molecules and can change through regulation of enzyme activity of LPS separate component synthesis. Regulation of gene expression, which determines LPS synthesis, can occur at a level of DNA primary structure (through mutations and recombinations); on highest levels of DNA organization through alteration of its conformation; can be mediated by signal transduction systems of bacterial cells. The changes of LPS structure can have spontaneous character due both to the instability of the appropriate genes and to the induction under the influence of various factors. Spontaneous variation of LPS structure is one of the reasons of heterogeneity of bacterial cell populations and creates a basis of their preadaptation. The influence of the environment factors on structure and functions of LPS can be both direct and mediated through regulation of expression of chromosome and plasmid genes by direct influence of the factors on DNA and through signal transduction systems of bacterial cells. Pathogenic bacteria can use the LPS variations at adaptation to biotic and abiotic factors. PMID- 15131974 TI - [Evolutionary development of the immunoglobulins super family]. AB - The origin of the ability of immunoglobulins (Igs) and T-cell receptors (TCRs) to specifically recognize antigens is related to the evolutionary development of proteins of the immunoglobulin superfamily (IgSF). The IgSF proteins are characterized by specific domain organization of molecules and statistically significant homology with known Igs. Four types of Ig domains (V1, V2, C1, and C2), differing from one another both in variations in their spatial organization and in the number of amino acid residues, have been distinguished. Immunoglobulin superfamily comprises Igs; TCRs; class I and II major histocompatibility complex (MHC) molecules; one-domain proteins of thymocytes and T-cells (Thy-1); myelin protein P0; beta 2-microglobulin; two-domain proteins--sponge receptor tyrosine kinase (RTK), sponge adhesive protein (SAP), Drosophila tyrosine-kinase receptor (DTKR), cortical-thymocyte receptors of Xenopus (CTX), human (CTH), etc.; and a large group of adhesins, coreceptors, and Ig receptors with varying number of domains. Evolutionary development of IgSF began with the origination of chaperones, Thy-1, and P0 of prokaryotes and unicellular eukaryotes. Mutations, duplications, and translocations of genes controlling both V and C domains yielded proteins differing in the number and combination of these domains. All IgSF proteins are divided into two groups. The first group includes the proteins with nonrearranging V2 domains and homophilic mode of interaction; the second, the proteins with rearranging V1 domains and heterophilic mode of interaction (Igs, TCRs). The acquisition of the capability of heterophilic antigen-binding mode of interaction was apparently related to the introduction of recombination activating retroviral genes (RAG1 and RAG2) into the genome of Gnathostomata ancestors. PMID- 15131976 TI - [Multiple forms of the proteasomes and some approaches to their separation]. AB - A technique for proteasome isolation from rat liver was developed; it allows isolation of 26S proteasome fraction free from 20S proteasome admixture. The technique includes consecutive stages of protein fractionation by different concentrations of ammonium sulfate, gel filtration on Sepharose 2B, and high speed ultracentrifugation. PMID- 15131975 TI - [Biochemical properties of the the lipoproteins lipid composition in fishes of the different ecology: trout Salmo irideus L. and whitefish Coregonus lavaretus L]. AB - We studied lipid composition of low- and high-density lipoproteins from serum of female rainbow trout Salmo irideus L. and whitefish Coregonus lavaretus L. Phospholipids are main lipids in all fractions and phosphatidylcholine is the major phospholipid. Species specificity of lipid fractions are due to ecological conditions typical for the studied fish species, a nearly domesticated rainbow trout and whitefish, a component of natural ichthyofauna. Serum of the studied fish species contains lipoprotein groups specific for other animals; however, high-density lipoproteins predominate in fish. PMID- 15131977 TI - [Study of the cholinesterase active site using a fluorescent probe]. AB - We studied fluorescence of 9-amino-1,2,3,4-tetrahydroacridine hydrochloride (tacrine) in the presence of serum cholinesterase. Quenching of tacrine fluorescence dependent on cholinesterase activity has been revealed. A quenching mechanism is proposed; it includes formation of a complex and charge transfer mediated by excited tacrine molecule as well as indole of tryptophan residue from the periphery of cholinesterase active site. PMID- 15131978 TI - [Effect of the toxic chemicals in low doses on phospholipid composition of the animal liver]. AB - We analyzed phospholipid composition in the liver 7, 14, and 31 days after single oral administration of oil solution of hexane-ether extracts from potable water sources with different content of benzo[a]pyrene and polychlorobiphenyls. Neither linear effect-dose relationship nor directed correlations between individual phospholipid fractions and generalized indices of their composition have been revealed, which indicates high sensitivity and no normalization of phospholipid composition in mouse liver one month after administration of toxicants. PMID- 15131979 TI - [Effect of ouabain, iodoacetamide, and pH on the human erythrocyte hemolysis induced by calcium ions in the presence of ionophore A23187]. AB - The effect of inhibitors of Na+, K(+)-ATPase (ouabain) and glycolysis (iodacetamide) as well as pH on calcium ion-induced erythrocyte hemolysis in the presence of ionophore A23187 is first described. Hemoglobin release decreases under the influence of ouabain, iodacetamide, and low pH, which is commonly observed at low temperature and in the samples studied in spring and summer. Active hemoglobin release through defects of the erythrocyte membrane under the influence of the transmembrane electrical potential was proposed to mediate hemolysis. PMID- 15131980 TI - [Calcium as a control factor of nitrogenase activity in Vicia faba L. root nodules: calcium release from symbiosomes and the accompanying decrease in their nitrogenase activity is blocked by verapamil]. AB - Treatment of root nodules or symbiosomes isolated from them with calcium chelator EGTA alone or with calcium ionophore A23187 for 3 h under microaerophilic conditions considerably decreased their nitrogenase activity (NA). Under these experimental conditions, cytochemical electron microscopy demonstrates a considerable calcium depletion in symbiosomes of the infected nodule cells by EGTA and A23187. Ca2+ channel blockers, verapamil and ruthenium red, inhibited EGTA-induced Ca2+ release from symbiosomes. In this case, NA insignificantly increased in the whole nodules and reached the baseline in symbiosomes. The experiments on isolated symbiosomes and arsenazo III demonstrated that verapamil inhibited Ca2+ transport induced by valinomycin at the background of K+ ions. These data suggest the presence of a verapamil-sensitive transporter on the peribacteroid membrane responsible for Ca2+ release from symbiosomes. A possible role of this transporter in the interaction between symbiotic partners in the infected cells of the root nodules is discussed. PMID- 15131981 TI - [Activation of the chloroplast respiration increases chlorophyll fluorescence yield in Chlorella adapted to darkness at higher temperature]. AB - A tenfold increase in chlororespiration during dark incubation of Chlorella perynoidosa Chick at high temperature doubled the initial chlorophyll fluorescence yield (F0). The presence of iodacetamide or unmetabolizable glucose analog 2-deoxy-D-glucose prevented increase in both chlororespiration and F0 yield. The rates of chlororespiration and F0 yield growth demonstrated a similar pattern of temperature dependence. Inhibition of electron transport between QA and plastoquinone prevented increase in F0 during dark respiration of the cells at high temperature. Apparently, a pool of plastoquinone was restored in the chlororespiratory chain during the dark incubation at 37.5-41 degrees C and plastoquinone exchanged electrons with QA. This is the cause of QA reduction and subsequent increase in F0 yield. PMID- 15131982 TI - [Principles of the acoustic communication between humans and birds]. AB - We analyzed the mechanism of alarm call generation in various bird species, species-specific calls in closely related species, and intraspecific differences in calls. Production of these signals proved to rely on common mechanism: variation of temporal indices within the frequency range limited by morphological structures of the vocal apparatus. These mechanisms underlie development of acoustic synthesizers mediating communication with birds and control of their behavior. The obtained data are discussed in terms of these signals processing by the auditory system of birds. PMID- 15131983 TI - [Involvement of the saiga liver in carbohydrate metabolism during ontogenesis]. AB - Histochemical investigation of saiga liver during ontogenesis was carried out. The dynamics of accumulation, expenditure, and localization of glycogen as well as neutral and acid glycoproteins was studied. Maximum accumulation of glycogen and neutral glycoproteins was observed during late fetal development, while maximum expenditure was specific for newborns. Localization of glycogen and glycoproteins was described. PMID- 15131984 TI - [Selective histochemical identification of neuronal cell populations using fucose specific lectins]. AB - We studied lectin histochemical properties of structures of caudal ganglia of the vagus nerve and ganglion of the trigeminal nerve in white rats using fucose specific conjugates to peroxidase. Morphological samples were processed on a computer video analyzer. Metrical and optical indices of the afferent neurons were analyzed. The obtained data demonstrate different topography of glycoconjugates in the afferent ganglia. Application of recent image processing techniques allows revealing neuron populations in afferent ganglia of rats undetectable by standard morphological techniques. PMID- 15131985 TI - [Stimulation of the delta 1-receptors help to decrease reperfusion induced myocardial stunning]. AB - In vivo or in vitro administration of a omega 1 receptor agonist d-SKF 10,047 (1 mg/kg intravenously or 10 mg/l in vitro) promoted an increase in the resistance of isolated perfused rat heart to ischemia/reperfusion injury. Both in vivo and in vitro stimulation of omega receptors prevents development of reperfusion contracture and release of creatine kinase and increases the developed pressure, double product, +dP/dt, and -dP/dt in the left ventricle. Activation of omega receptors has no significant effect on the occurrence of reperfusion arrhythmias ex vivo. Stimulation of cardiac sigma receptors is proposed to prevent myocardial stunning. PMID- 15131987 TI - [Radiocarbon dating of pollen and spores in wedge ice from Iamal and Kolyma]. AB - Radiocarbon dating of pollen concentrate from late Pleistocene syngenetic wedge ice was carried out using acceleration mass spectrometry (AMS) in Seyakha and Bizon sections. Comparison of the obtained dating with palynological analysis and AMS radiocarbon dating previously obtained for other organic fractions of the same samples allowed us to evaluate accuracy of dating of different fractions. Quantitative tests for data evaluation were considered in terms of possible autochthonous or allochthonous accumulation of the material on the basis of pre Pleistocene pollen content in these samples. Paleoecological information content of pollen spectra from late Pleistocene syngenetic wedge ice was evaluated. PMID- 15131986 TI - [Extracellular carbohydrates and polysaccharides of the algae Chlorella pyrenoidosa Chick S-39]. AB - In the growing culture of the thermophilic alga Chlorella pyrenoidosa Chick S-39, the amount of extracellular carbohydrates in the medium reached 5-17% of their content in the cells and 20-40% of the total content of extracellular organic matter. Experiments with the enrichment and synchronous algal cultures showed that the accumulation of extracellular carbohydrates and polysaccharides in the media occurred due to their release from the cells, rather than to cell lysis, and depended on cell photosynthetic activity and reproduction. Chromatographic determination of free sugars revealed the presence of saccharose, glucose, and fructose in the culture medium. Extracellular carbohydrates in C. pyrenoidosa cultures were represented mainly by water-soluble polysaccharides containing galactose, mannose, arabinose, xylose, ribose, fucose, and rhamnose. PMID- 15131988 TI - [Spatiotemporal dynamics of the vole populations in heterogeneous habitats of the Iremel mountain range]. AB - Specific features of seasonal changes in the demographic structure and distribution of three vole species were studied in heterogeneous, qualitatively different habitats of three altitudinal belts in the Southern Ural. The results showed that the fauna of small mammals in the mountain-forest, subgoltsy, and mountain-tundra belts of the Iremel' Mountain Range could be regarded as metapopulations of the northern red-backed vole, large-toothed red-backed vole, and root vole, and the fauna of each belt could be regarded as micropopulations or local populations of the same species. Statistically significant differences in the population dynamics and demographic structure of voles inhabiting different biotopes were revealed. PMID- 15131989 TI - [Nitrogen pool in northern taiga larch forests of Central Siberia]. AB - The pools of nitrogen in different blocks of forest ecosystems and its cycle in the soil are considered. It is shown that the bulk of nitrogen concentrates in the soil and dead organic matter (necromass) of an ecosystem. The nitrogen pool of forest litters and soils consists by 83-93% of the inert compounds that cannot be involved in the biological cycle. Mineralization of organic nitrogen containing substances in the litters and soils usually yields ammonium as an end product. The amount of nitrogen mineralized over the growing season is partially expended for annual plant increment (30-65%) and immobilization (12-17%), with its large proportion being found in the soil. PMID- 15131990 TI - [New considerations in the treatment of functional dyspepsia]. AB - Functional dyspepsia is a clinical syndrome defined by chronic or recurrent pain or discomfort in the upper abdomen of unknown origin. Dyspepsia is a very common condition, with a high prevalence in the general population. The multifactorial pathogenesis of dyspeptic symptoms is poorly defined. However, several factors have been identified as relevant, including the role of gastric acid secretion, gastroduodenal dysmotility, visceral hypersensitivity and also the effects of stress and psychological factors. The role of Helicobacter pylori infection is more controversial, the infection plays only an irrelevant role in the pathophysiology of functional dyspepsia, nevertheless in a subgroups of patients eradication therapy is generally accepted as a preventive tool. The need for positive diagnosis is emphasized. The management of dyspeptic patients has a considerable impact on health services. As the costs of initial investigation are high, most patients receive empirical management (prescribing without a proven diagnosis), at least initially. Subgrouping of functional dyspepsia according to its predominant symptom into ulcer-like, dysmotility-like and non-specific dyspepsia has been attempted to tailor its treatment accordingly. This review aims to summarize the new aspects of pathophysiology, investigation and management strategies of functional dyspepsia. PMID- 15131991 TI - [Diabetes mellitus and tumors of the oral cavity--epidemiologic correlations]. AB - INTRODUCTION AND AIMS: Numerous publications have already demonstrated that diabetes mellitus (DM) is a risk factor for the development of periodontal diseases and various inflammatory lesions in the oral mucosa. A possible correlation between DM and oral premalignancies and tumors was examined in this study, as no literature data are available concerning this problem. METHODS: Stomato-oncological screening was carried out on 200 DM patients in the medical departments: The lesions found were classified in three groups: inflammatory lesions, benign tumors and precancerous lesions. Benign tumors were found in 14.5%, and precancerous lesions in 8% of diabetics. Earlier Hungarian screening studies indicated lower frequency of these lesions in the general population. A retrospective DM screening of 610 inpatients with histologically confirmed oral malignancies was also performed. The control group comprised 574 complaint- and tumor-free adults. Fasting blood glucose levels were determined in both groups, and the tumor location was registered in the cancer patients. RESULTS: In the group of the oral cancer patients, DM was present in 14.6% and an elevated blood glucose level in 9.7%. These values are significantly higher than those for the tumor-free control group (p < 0.01). The gingival and labial tumor location was significantly more frequent among diabetic cancer patients than in the non diabetic group (p < 0.01). CONCLUSION: The combination of DM and smoking means a higher risk for oral precancerous lesions and malignancies. DM may be a risk factor for oral premalignancies and tumors. PMID- 15131992 TI - [Methods of monitoring oxidation-reduction balance and its potential role in diagnostics]. AB - Results of the latest investigations confirm that the injury of redox homeostasis of human organism can be the starting point of many diseases. The pathogenesis of these diseases, including atherosclerosis, diabetes mellitus, chronic liver diseases, renal failure, rheumatoid arthritis and neurodegenerative diseases could be cleared up more deeply by the investigation of parameters of oxidative stress and by the observation of their changes in connection with the different organ and tissue damages. It is well known, that individuals with lowered antioxidant defences may be at greater risk of developing diseases induced by free radicals. The goal of the authors is to direct attention to the possible role of some redox parameters in detecting of general health status of the human body and in the risk assessment of the different diseases. These measurements can perhaps help the clinician in determining optimal treatment and monitoring its effectiveness. PMID- 15131993 TI - [Scientific communication at the beginning of the 21st century--Hungarian survey and international outlook in medicine and the life sciences]. AB - INTRODUCTION: Rapid developments take place in communication technology. It is a hard task to keep pace with them, but unavoidable. The scene of electronic communication and publication, the Internet has originally been constructed for scientific communication and publication of scientists. Nowadays, the electronic communication has spread all over the world. Though the share of sciences has been diminished, the progress in the scientific electronic communication has not been broken. AIM: The authors delineate the present situation in electronic communication in general and in Hungary. METHODS: The authors asked by questionnaire survey the medical scientists and professionals about their knowledge and usage of modern publication possibilities. For comparison's sake, the authors referred to some foreign surveys as well. RESULTS: In Hungary, the communication possibilities on the Internet--and the electronically accessible publications--are at the readers' disposal at the universities and academic institutions. The opinions about the popularity and effects of electronic publication are generally positive. Concerns were expressed in connection with the possible weaker professional control in case of certain types of publication. In the opinion of the majority of questioned people the electronic form of publication will never replace the traditional one completely, it will be, however, dominant. PMID- 15131994 TI - [Recommended therapy in chronic obstructive lung disease]. PMID- 15131995 TI - [Health care of the Rakoczi freedom fight]. PMID- 15131996 TI - [In memory of Jozsef Antall, Prime Minister of Hungary--on the tenth anniversary of his death]. PMID- 15131997 TI - The care and feeding of critical incident stress management. AB - Critical Incident Stress Management (CISM) is under attack by some major academic and training institutions. Despite its acceptance internationally, CISM is viewed by some as "dangerous and damaging." The author's recent experience in a number of mental health training workshops on primary care of September 11-related dysfunction is reviewed, along with arguments in support of the CISM model of crisis intervention. A proposal for greater efforts at empirical foundations is offered. PMID- 15131998 TI - The effectiveness of critical incident stress debriefing with primary and secondary trauma victims. AB - Critical incident stress debriefing (CISD) (Mitchell, 1983) has been under intense scrutiny recently in the psychology literature. Several authors have concluded that CISD has no effect or even negative effects on victims of trauma (Bisson, McFarlane, & Rose, 2000; van Emmerik, Kamphuis, Hulsbosch, & Emmelkamp, 2002). This review calls these conclusions into question by critically contrasting: (1) studies that utilize CISD with emergency services personnel ("secondary" victims of trauma, for whom the CISD and the larger Critical Incident Stress Management models were first created) and (2) studies that utilize CISD with primary victims of trauma. This review suggests that CISD is an effective method of reducing risk for PTSD-related symptoms in emergency services personnel. However, when debriefings are conducted with primary victims of traumatic events (e.g., accident victims, burn victims still in the hospital), the results are much less promising. The authors conclude that protocols are needed for interventions with primary victims to help mitigate the impact of trauma on this population. Appropriate interventions might include risk assessment and appropriate referral services, or in some instances, focused psycho-educational group debriefings. Further research is clearly warranted to examine the effectiveness of alternative interventions for the primary victim population. PMID- 15131999 TI - Gender related correlates of posttraumatic stress symptoms in a World Trade Center tragedy sample. AB - Previous research has documented that women are more susceptible to developing posttraumatic stress disorder than men. This study examines the relationships between gender, ruminative coping, traditional gender role characteristics, trauma-related cognitions, and posttraumatic stress symptoms. Participants were 123 victims of the September 11, 2001, terrorist attacks, including 72 males and 51 females. Results indicated that female victims had more posttraumatic stress symptoms than male victims. Ruminative coping was positively correlated with posttraumatic stress symptoms. Victims who were categorized as 'feminine' had significantly higher levels of posttraumatic stress symptoms than victims categorized as 'androgynous.' Multiple regression and path analyses were used to determine whether ruminative coping, traditional gender role characteristics, and trauma-related cognitions mediated the gender difference in posttraumatic stress symptoms. The mediational model was not supported by these data. PMID- 15132000 TI - Coping with traumatic loss: an interview with the parents of TWA 800 crash victims and implications for disaster mental health professionals. AB - The stories of survivors of traumatic events can be instructive to professionals who provide crisis intervention and/or psychotherapy to the bereaved. This paper provides excerpts from an interview with a middle-age married couple who lost two children in the 1996 explosion of TWA 800 over Long Island Sound. Their story illustrates important methods of coping with grief and sheds light on the value to survivors of recovering victims' remains. This case study is followed by a discussion about some elements of the interview that are helpful to an understanding of the grief process. PMID- 15132001 TI - Death notification: considerations for law enforcement personnel. AB - Death notification is among the most unpleasant and uncomfortable tasks a law enforcement officer undertakes. For years, law enforcement officers have been performing notifications that involved life threatening injuries or death from tragedies including suicide. Unfortunately, many officers have performed this task without appropriate training. Performing notifications can be physically and emotionally exhausting. Expressing the right words, understanding families' emotions, and responding with empathy are paramount. The attitude, demeanor, professionalism, and sensitivity of law enforcement officers projected during a notification are critical. The delivery of a notification can remain etched in a family member's memory forever. Many notifications have left family members with the perception that law enforcement officers are callous, thoughtless, and insensitive. This negative perception can be overcome through training and practice. This article presents several factors that supervisors and officers should consider in the notification process and offers guidelines to consider to best serve your agency and the community. PMID- 15132002 TI - The psychologist's role in hostage negotiations. AB - The purpose of this article is to examine the proper role of the psychologist who serves as a consultant to a hostage negotiation team. Mental health consultants can be beneficial to law enforcement during these times of crisis if they are aware of the appropriate role they play. This article discusses preferred characteristics of a mental health professional serving in this capacity and focuses on the various tasks required of a psychologist on the scene of a barricade. PMID- 15132003 TI - [Role of neuroglia in central nervous system diseases]. AB - Glia form a cellular network consisting mainly of astrocytes that nourish the neurons, oligodendrocytes that form myelin, and microglia that ensure the immunological defence of the nervous system. Experimental allergic encephalomyelitis is an animal model of multiple sclerosis, characterised by cellular infiltrates and demyelinisation in the central nervous system. It can be induced in susceptible rats (DA) by inoculation of foreign protein. Other, non susceptible rats (AO) have infiltrates that disappear without functional signs of disease. We have found microglia and astrocytes in infiltrates 10 days after inoculation. At 14 days they are even more marked in the DA, but astrocytosis is less in the AO. Microglia have disappeared at 14 days in the DA, but persist in the AO. We suggest that the infiltrates are dispersed by microglial activity that persists in the AO and prevents astrocytosis. In the DA the loss of microglia allows gliosis to occur, with demyelination due to lesions of oligodendrocytes, and clinical signs. We have also studied astrocytes and microglia in the cerebral cortex of schizophrenics and control non-schizophrenics. The number of astrocytes does not vary between patients and controls, but microglia are more numerous in the patients. We conclude that there is a cortical microgliosis in schizophrenia that may be a response to an earlier lesion, perhaps protecting neighbouring neurons. PMID- 15132004 TI - ["SHIP2 and insulin signaling: what is its role in the pathogenesis and treatment of type 2 diabetes?"]. AB - Based on the analysis of SHIP2+/- and SHIP2-/- mice showing that the lipid phophatase SHIP2 is an important negative regulator of the insulin pathway, we have suggested that the enzyme is a candidate gene for genetic predisposition to type 2 diabetes, and a potential therapeutic target for the same disease. Two years after, we propose to review these hypothesis in light of recent results from our laboratory, from international collaborations and from competitors in the field. PMID- 15132005 TI - [Academic praise to Professor J. Radermecker]. PMID- 15132006 TI - [A new mechanism of tumor resistance to the immune system, based on tryptophan breakdown by indoleamine 2,3-dioxygenase]. AB - T lymphocytes represent the main effectors of the immune response that can lead to tumor rejection, which represents the aim of various approaches of immunotherapy that are currently tested. However, in many cases, tumor cells appear to resist immune rejection. We have recently uncovered a new mechanism of tumoral immune resistance based on the expression by tumor cells of indoleamine 2,3-dioxygenase (IDO), an enzyme that rapidly degrades tryptophan, an amino acid that is crucial to sustain proliferation of T lymphocytes. We showed that most human tumors constitutively express IDO. We also observed that expression of IDO by immunogenic mouse tumor cells, prevents their rejection by pre-immunized mice. This effect is accompanied by a lack of accumulation of specific T cells at the tumor site, and can be partly reverted by systemic treatment of mice with an inhibitor of IDO, in the absence of noticeable toxicity. These results suggest that the efficacy of therapeutic vaccination of cancer patients might be improved by concomitant administration of an IDO inhibitor. PMID- 15132007 TI - Cultural considerations and issues in measurement. PMID- 15132008 TI - Metric equivalence assessment in cross-cultural research: using an example of the Center for Epidemiological Studies--Depression Scale. AB - Metric equivalence is a quantitative way to assess cross-cultural equivalences of translated instruments by examining the patterns of psychometric properties based on cross-cultural data derived from both versions of the instrument. Metric equivalence checks at item and instrument levels can be used as a valuable tool to refine cross-cultural instruments. Korean and English versions of the Center for Epidemiological Studies-Depression Scale (CES-D) were administered to 154 Korean Americans and 151 Anglo Americans to illustrate approaches to assessing their metric equivalence. Inter-item and item-total correlations, Cronbach's alpha coefficients, and factor analysis were used for metric equivalence checks. The alpha coefficient for the Korean-American sample was 0.85 and 0.92 for the Anglo American sample. Although all items of the CES-D surpassed the desirable minimum of 0.30 in the Anglo American sample, four items did not meet the standard in the Korean American sample. Differences in average inter-item correlations were also noted between the two groups (0.25 for Korean Americans and 0.37 for Anglo Americans). Factor analysis identified two factors for both groups, and factor loadings showed similar patterns and congruence coefficients. Results of the item analysis procedures suggest the possibility of bias in certain items that may influence the sensitivity of the Korean version of the CES D. These item biases also provide a possible explanation for the alpha differences. Although factor loadings showed similar patterns for the Korean and English versions of the CES-D, factorial similarity alone is not sufficient for testing the universality of the structure underlying an instrument. PMID- 15132009 TI - Assessing the cultural appropriateness of the Finding Meaning Through Caregiving Scale for Korean caregivers. AB - Before psychometric instruments can be used for populations other than those for whom they were originally developed, validation of cultural appropriateness is essential. This article describes the assessment of the cultural appropriateness of the Finding Meaning Through Caregiving Scale (FMTCS) with Korean female family caregivers. The FMTCS measures finding meaning among caregivers from an existential perspective and has three subscales: Loss/Powerlessness, Provisional Meaning, and Ultimate Meaning. The instrument's cultural appropriateness was examined through semistructured interviews with ten Korean-born female family caregivers, five caregivers living in Korea, and five living in the United States. The interview data are reported according to the three dimensions described by Flaherty and colleagues (1988): content, semantic, and conceptual equivalence. Although the majority of items of the FMTCS appeared applicable to Korean caregivers, items on the Loss/Powerlessness and Provisional Meaning subscales asking caregivers about feelings related to missing their past relationships or communications appeared inappropriate for many Korean daughters in-law. Of equal importance, a unique source of meaning among Korean caregivers that is not assessed in the FMTCS is an interpersonal context, including the importance caregivers place on teaching children and feeling proud of one's caregiving accomplishments in the eyes of other relatives. The addition of new items that address the interpersonal context is warranted to improve the instrument's cultural appropriateness for Korean caregivers. PMID- 15132010 TI - Development of a Cultural Competence Assessment instrument. AB - This article describes initial testing of an instrument designed to provide evidence of cultural competence among health care providers and staff. The Cultural Competence Assessment (CCA) instrument was based on a model describing cultural competence components (fact, knowledge, attitude, and behavior). Content and face validity were confirmed through expert panel review, subject feedback, and field-testing. The CCA was administered to an interdisciplinary health care team in a community hospice setting. Preliminary findings suggest that the CCA performed well. Internal consistency reliability for the scale was 0.92. Construct validity by factor analysis demonstrated that 25 items had loadings above 0.42. Construct validity was supported with a significant correlation to the widely used Inventory for Assessing the Process of Cultural Competence among Health care Professionals (IAPCC). Validity also was supported by significant differences between individuals with different educational levels and prior diversity training. The CCA is a promising tool to measure cultural competence in populations with a wide range of educational levels and backgrounds. PMID- 15132011 TI - Culturally appropriate family assessment: analysis of the Family Assessment Device in a pediatric Chinese population. AB - As part of a larger study, we investigated the adaptation of the Family Assessment Device (FAD) to a Chinese population of hospitalized children (N = 313) compared to a sample of families with healthy children (N = 29) in Hong Kong and Chinese Mainland. Confirmatory factor analysis and exploratory factor analysis (EFA) were performed to examine the FAD structure. The results supported the notion of cultural variations in measuring family functioning. Eight factors were found to explain 30.34% of the variance in family functioning. The Cronbach's alphas of families with hospitalized children ranged from 0.29 to 0.74. Similar reliability scores were found in nonhospitalized families. This study indicates that the Chinese FAD has a different factor structure, reliabilities, and mean scores in several subscales compared to U.S. studies. The psychometric properties of the Chinese FAD may be influenced by the fact that the sample was focused on children rather than adults. PMID- 15132012 TI - Psychometric properties of the Spanish version of the Resilience Scale. AB - The purpose of this study is to test the reliability and validity of a Spanish translation of the Resilience Scale (RS), which was originally created in English by Wagnild and Young (1993). A team of bilingual, bicultural translators participated in the translation process to enhance the linguistic accuracy and cultural appropriateness of the Spanish translation. As part of the convenience sample of 315 women of Mexican descent who participated in the larger study, data from 147 women who preferred to read and write in Spanish were used in this analysis. The English version of the RS consists of a 17-item "Personal Competence" subscale and an 8-item "Acceptance of Self and Life" subscale for a total of 25 items. However, two items had low item-total loadings and were removed to form a modified 23-item RS. The exploratory principal components factor analysis, varimax rotation, and subsequent goodness of fit indices were ambivalent on whether a one or two-factor solution was appropriate, but the chi square difference test clearly demonstrated that the two-factor solution of the Spanish version was more useful in explaining variance than a one-factor solution. Internal consistency reliability was estimated with Cronbach's alpha (alpha = 0.93) which was acceptable for the 23-item RS as well as its subscales. Construct validity was demonstrated by a significant positive correlation between resilience and life satisfaction (r = 0.36; p < 0.001), and a significant negative correlation between resilience and depressive symptoms (r = -0.29; p < 0.01). This analysis ultimately supports the appropriateness of the modified 23 item Spanish translation of the RS and its subscales in a sample of urban, low income women of Mexican descent in the U.S. PMID- 15132013 TI - Comparison of blood pressure measurement consistency using tonometric and automated oscillometric instruments. AB - To effectively evaluate treatments for hypertension, researchers and clinicians must be able to measure blood pressure (BP) in a valid and reliable way. The purpose of this study is to compare measurements made in the clinic using beat-to beat radial BP tonometry, measurements made during 24 hours using an ambulatory BP monitor, and measurements made in the clinic using an automated oscillometric BP monitor. Fifty-seven adults with primary hypertension participated in this study, which used a repeated measures descriptive design. Clinic and ABPM daytime averages were compatible for both SBP and DBP. In contrast, clinic SBP was 7.56 mmHg higher than the beat-to-beat SBP; clinic DBP was 9.83 mmHg higher than the beat-to-beat DBP. These data suggest that automated clinic measurements may be used in place of daytime ambulatory BP measurements. We also estimate sample sizes for future studies based on characteristics of clinic BP. PMID- 15132014 TI - No child left behind: the good, the bad, and the ugly. PMID- 15132015 TI - Comparing practical knowledge storage of deaf and hearing teachers of students who are deaf or hard of hearing. AB - Especially in the education of students who are deaf or hard of hearing, teachers' practical knowledge storage is almost never measured. The Survey of Practical Knowledge was used to compare the practical knowledge storage of deaf and hearing teachers of these students. Surveyed were 48 deaf and 115 hearing individuals at the preservice and in-service experience levels. Practical knowledge storage was defined as images, rules of practice, and practical principles. Results indicate that deaf teachers tend to view students as equals but are more likely to emphasize control over classroom behavior than hearing teachers. Hearing teachers tend to stress efforts to engage students in subject matter by providing variety and relating it to life experiences. Given the trend toward high-stakes testing of teachers, further research is encouraged on role differences between deaf and hearing teachers working with students who are deaf or hard of hearing. PMID- 15132016 TI - W(h)ither the deaf community? Population, genetics, and the future of Australian sign language. AB - According to enrollments in schools for the deaf and data from the national census and neonatal hearing screening programs, the incidence of severe and profound childhood deafness in Australia is, and has been, less than commonly assumed. Factors implicated include improved medical care, mainstreaming, cochlear implants, and genetic science. Data for the United States, Britain, and other developed countries seem consistent with those for Australia. Declining prevalence and incidence rates have immediate implications for sign-based education, teacher-of-the-deaf training programs, and educational interpreting. There are also serious consequences for research, documentation, and teaching regarding Australian Sign Language (Auslan), and for the future viability of Auslan. Prompt action is essential if a credible corpus of Auslan is to be collected as the basis for a valid and verifiable description of one of the few native sign languages in the world with significant attested historical depth. PMID- 15132017 TI - Deaf persons and computer use. AB - Deaf person's computer use was studied (N = 227). Respondents self-administered a survey in their preferred language (voice, American Sign Language, captions, or printed English). A small nonparticipant sample was also recruited. Demographics were consistent with those in other studies of deaf people: 63% of respondents reported computer use, mostly at home; 50% of nonparticipants reported computer use. Subjects with hearing loss due to meningitis were less likely to use computers (p = .0004). Computer use was associated with English usage at home (p = .008), with hearing persons (p = .002), and with physicians and nurses (p = .00001). It was also associated with the use of Signed English as a child to communicate (p = .02), teacher use of Signed English (p = .04), and teacher use of ASL (p = .03). Two thirds of respondents reported using computers, though nonresponder data suggested less use among all deaf persons. Computer use was associated with English use and inversely associated with hearing loss due to meningitis. PMID- 15132018 TI - Measuring the attitudes of human service professionals toward deafness. AB - The Attitudes to Deafness Scale is a 22-item measure of attitudes toward people who are deaf designed for use with human service professionals. Attitude statements were generated from personal accounts by deaf people in the literature and from a focus group in which deaf people discussed their experience of hearing people's attitudes toward them. A 60-item scale was administered to a group of 121 clinical and forensic psychologists during their training. Item analysis was conducted to select items that effectively distinguished participants with a positive attitude from those with a negative attitude toward deaf people. The scale may be used in any context where a professional group comes into contact with people who are deaf. PMID- 15132019 TI - Mental health problems of deaf Dutch children as indicated by parents' responses to the child behavior checklist. AB - Emotional/behavioral problems of 238 deaf Dutch children ages 4-18 years were studied. Parental reports indicated that 41% had emotional/behavioral problems, a rate nearly 2.6 times higher than the 16% reported by parents of a Dutch normative sample. Mental health problems seemed most prevalent in families with poor parent-child communication. Deaf children ages 12-18 showed more problems with anxiety and depression and more social problems than those ages 4-11. Deaf children with relatively low intelligence showed more social problems, thought problems, and attention problems than those with relatively high intelligence. The authors stress the need to get information on deaf children's mental health functioning not just from parents but from other informants such as teachers and the children themselves. An expansion assessment of deaf children, and of special services and treatments for deaf children and adolescents with emotional/behavioral problems, is recommended. PMID- 15132020 TI - Making public mental-health services accessible to deaf consumers: Illinois Deaf Services 2000. AB - Illinois Deaf Services 2000 (IDS2000), a public/private partnership, promotes the creation and implementation of strategies to develop and increase access to mental health services for deaf, hard of hearing, late-deafened, and deaf-blind consumers. IDS2000 has resulted in the establishment of service accessibility standards, a technical support and adherence monitoring system, and the beginnings of a statewide telepsychiatry service. These system modifications have resulted in increase by 60% from baseline survey data in the number of deaf, hard of hearing, late-deafened, and deaf-blind consumers identified in community mental-health agencies in Illinois. Depending on the situation of deaf services staff and infrastructure, much of IDS2000 could be replicated in other states in a mostly budget-neutral manner. PMID- 15132021 TI - Data-driven assessment of teacher candidates during their internships in deaf education. AB - Deaf education teacher training programs have received criticism for graduating students with gaps in knowledge and skills required for specific placements. The National Council for Accreditation of Teacher Education (NCATE) requires that accredited programs engage in self-study, and NCATE guidelines require programs to collect valid, reliable evidence of teacher candidate performance and candidates' effect on student learning. In the present study, an ecobehavioral assessment computer program, MS-CISSAR (Mainstream Code for Instructional Structure and Student Academic Response), was used in evaluations of 8 teacher candidates during internship experiences. Results were mixed. Some of the instructional arrangements, teaching behaviors, and student responses resembled those found in studies using in-service teachers as subjects; however, some data revealed a need for changes in instruction. Data gathered with MS-CISSAR could be added to evaluations to help training programs meet NCATE requirements for evidence on teacher candidate performance and candidates' effect on student learning. PMID- 15132022 TI - A view from outside. PMID- 15132023 TI - Goal-directed upper limb movements by children with and without DCD: a window into perceptuo-motor dysfunction? AB - BACKGROUND AND PURPOSE: Developmental co-ordination disorder (DCD) occurs in at least 6% of school-aged children. Researchers agree that motor co-ordination problems evident in DCD are, in part, the result of perceptual and cognitive processes, but the limited research available remains inconclusive. The present study investigated perceptual-motor abilities, with regard to vision, kinaesthesia and cross-modal judgement, in children with and without DCD. METHOD: A cross-sectional study design was used. Nine children, aged six years (+/- six months) with DCD, and nine children without DCD, matched for age and gender, participated in the study. The children were required to point with the preferred hand to a target in three different positions under four sensory conditions, either with or without vision. Three-dimensional motion analysis was used to investigate trajectory lengths, endpoint error and movement time. The results were analysed using a generalized linear mixed model to examine the systematic effects of group, target position and task. RESULTS: Compared with children without DCD, the children with DCD produced larger endpoint errors, greater movement times and longer trajectories. Children in both groups produced larger endpoint errors, greater movement times and longer trajectories in non-visually guided aiming versus visually guided aiming tasks. CONCLUSIONS: Children with DCD moved more slowly, with longer movement trajectories and were less accurate than children without DCD when aiming to all target positions under all sensory conditions. The greatest error and trajectory length occurred for both groups when aiming movements were performed in the absence of vision. As children in the DCD group had difficulties with movement executed under kinaesthetic or visual control, the results indicate that the normal advantage of vision displayed by children without DCD is not apparent, and visual and kinaesthetic problems may be present in children with DCD. PMID- 15132024 TI - Attitudes and beliefs of Brazilian and Australian physiotherapy students towards chronic back pain: a cross-cultural comparison. AB - BACKGROUND AND PURPOSE: The attitudes and beliefs of physiotherapists and students can potentially influence the outcome of treatment of low back pain. These attitudes and beliefs may be influenced by external factors, such as ethnicity. No study that compared the attitudes and beliefs of physiotherapy students from different cultural backgrounds, such as from Brazil or Australia, towards chronic low back pain was found. The purpose of the present study was therefore to compare the attitudes and beliefs of Brazilian physiotherapy students with those of Australian physiotherapy students and to published data from North American healthcare providers, and to investigate whether a history of chronic low back pain affects students' attitudes and beliefs. METHOD: A survey study design was used. Data were collected from 153 Brazilian physiotherapy students and compared with existing data from Australian physiotherapy students who had never been exposed to a chronic low back pain teaching module. Students' attitudes and beliefs were assessed by use of the HC-PAIRS questionnaire. These data were also compared with published data from North American healthcare providers. RESULTS: The Brazilian physiotherapy students had significantly higher scores on the HC-PAIRS questionnaire than the Australian students and the North American healthcare providers. A previous history of chronic low back pain did not affect students' attitudes and beliefs. CONCLUSIONS: As demonstrated by higher HC-PAIR scores, the Brazilian physiotherapy students agree more strongly with the notion that low back pain justifies disability and activity limitation than do Australian physiotherapy students and North American healthcare providers. A history of chronic low back pain does not affect students' attitudes and beliefs. PMID- 15132025 TI - Effect of problem-based learning on stages of change for exercise behaviour in patients with coronary artery disease. AB - BACKGROUND AND PURPOSE: Physical activity plays an important part in preventing coronary artery disease and is targeted in most rehabilitation and education programmes. The aim of the present study was to evaluate stages of change for exercise behaviour in patients with a recent event of coronary artery disease. METHOD: A randomized two-group, pre--post-test design was used. Fifty-seven subjects were included in the problem-based learning intervention group and 61 subjects were included in the traditional rehabilitation control group. Data were analysed by intention-to-treat. A single-item five-category scale, based on the 'Stages of Change' model was used to measure the level of exercise behaviour. The statistical analysis used two non-parametric approaches for ordered categorical data. RESULTS: There was a significant systematic change over time towards the extreme scale categories in both groups. This suggested that individuals who were inactive before the intervention regressed, whereas individuals with some interest in physical activity remained static or improved. No significant differences between groups were found. CONCLUSION: Problem-based learning did not significantly influence patients' progression through exercise behaviour stages. Rehabilitation teams should be observant of inactive participants and their greater risk of regression. PMID- 15132026 TI - Head stabilization strategies in the sagittal plane during locomotor tasks. AB - BACKGROUND AND PURPOSE: Head stability is the dynamic process of maintaining an equilibrium position of the head-in-space. Individuals with vestibular deficits restrict head movements during dynamic activities in an effort to adapt to vestibular loss. However, this strategy does not provide them with a successful means for adaptation during dynamic tasks where head movements are required. Therefore, identification of successful head stabilization strategies is the first step towards improving the rehabilitation of these patients. The purpose of the present study was twofold: to characterize the sagittal plane head stabilization response during walking; and to identify successful head stabilization strategies during normal walking and during a walking task that challenged head stability. METHOD: The study used a repeated-measures design. Eight healthy volunteers walked normally (normal condition) and walked whilst swinging their arms at twice the natural frequency (frequency condition). Head and trunk angular velocities were measured to determine head velocity magnitudes, and head-on-trunk, with respect to trunk gains and phases across the frequency spectrum of walking. RESULTS: The frequency condition increased the challenge to head stabilization and produced phases indicative of increased head stability (p < 0.05). Post hoc analyses revealed that the shift in phase occurred at higher frequencies (> 8 Hz) (p < 0.05). Increased head velocity magnitudes (p < 0.05) accompanied by decreased variability (p < 0.05) were also found at higher frequencies for the frequency condition. CONCLUSIONS: The results were indicative of a tightly controlled movement strategy that ensured head stabilization under conditions where head stability was challenged. This strategy was characterized by head-on-trunk movement that was equal and opposite to trunk motion. PMID- 15132027 TI - Variability in pain intensity, physical and psychological function in non-acute, non-traumatic neck pain. AB - BACKGROUND AND PURPOSE: Variability in clinical presentation among patients with neck pain disorders, in relation to symptom severity and the effect of the disorder on their physical and psychological function, has not been well documented. The present study examined the variability in pain intensity, physical and emotional or cognitive function in patients with non-acute, non traumatic neck pain, and the extent to which these factors are associated at the time of referral for physiotherapy treatment. METHOD: The study utilized a survey of patients referred for physiotherapy treatment for non-acute, non-traumatic neck pain. Seventy-two patients (mean age 58 years; SD 15.5 years) with a symptom duration ranging from four weeks to 30 years were recruited. Each completed measures of pain intensity using a visual analogue scale, and neck-pain related disability by use of the 'Neck Pain and Disability' scale (NPAD). Frequency distributions were calculated to examine variability in pain intensity, the total NPAD score and the effect of the disorder on different aspects of physical and emotional or cognitive function. Relationships between pain intensity, and physical and psychological function, were also examined. RESULTS: Pain intensity was normally distributed with a mean score of 6.3 cm (SD 2.5 cm). NPAD scores were normally distributed (mean 54.8%; SD 18%). Similar variability was noted in each of the NPAD factor scores for neck function, pain intensity, emotional or cognitive function and activities of daily living. Pain intensity was significantly (p < 0.01) correlated with the total NPAD score (r = 0.55) and with each of the four effects of neck pain represented in the NPAD sub-domains (r = 0.42-0.63). Symptom duration was not significantly correlated with pain intensity, or with the severity of disability, as reflected in the total NPAD score. CONCLUSIONS: The results highlight the variability between patients with neck pain, in relation to pain intensity and the effect of the disorder on physical and psychological function. Identifying the specific effects of the neck pain disorder in individual cases is important when defining the treatment objectives and selecting measures of treatment response. PMID- 15132028 TI - Living her dream. PMID- 15132029 TI - Apartheid. A decade on. PMID- 15132030 TI - Break the silence. PMID- 15132031 TI - Caring partnership. PMID- 15132032 TI - Age concern. PMID- 15132033 TI - Value real-time research. PMID- 15132034 TI - Help needed inside. PMID- 15132035 TI - Developing a palliative care outreach service. AB - This article describes the development of a palliative care outreach service by two nurse practitioners into ten community hospitals. The service was designed to improve equity of service delivery in the area and to meet national initiatives aimed at developing palliative care services. It involved responding to direct referrals while developing an educational programme for staff members. The results of a survey, which identified achievements for patient care and staff development, are discussed. PMID- 15132036 TI - Strategies to help students learn effectively. AB - All qualified nurses are involved in teaching and learning. This article presents four vignettes of adult learners training to be nurses and examines the factors that may be inhibiting their learning. It goes on to describe some of the strategies and approaches that might help these students to learn more effectively, including the use of learning contracts and reflective practice. PMID- 15132037 TI - Huntington's disease. AB - Huntington's disease is a complex degenerative disorder that affects the central nervous system. Although it is a rare condition, nurses are ideally placed to assess and manage patients with the disease, while also providing information and support to family members. This article discusses the cause and symptoms, explains the method of diagnosis and outlines the role of the nurse in caring for patients with Huntington's disease. PMID- 15132038 TI - Pressure ulcer incidence reporting. AB - AIM: To examine the accuracy of data collected on pressure ulcer incidence. A system for monitoring incidence has been in place since 1991 at a medium-sized hospital trust. The data collected are used to calculate resource requirements for specialist mattresses, to audit compliance with pressure ulcer guidelines and to identify staff training needs. METHOD: A convenience sample comprising 211 inpatients from the medical, surgical, rehabilitation and orthopaedic wards was assessed by the researcher over a 14-day period. Ward staff continued their usual routine of assessment and treatment. At the end of the study period, the researcher-generated data and ward staff-generated data were compared. RESULTS: A total of 343 assessments were carried out on 211 patients; 152 patients were found to have, or be at risk of having, pressure ulcers. The main finding after comparing both sets of data was that a significant number of patients at risk of, or with, pressure ulcers were not reported as such by ward staff (n = 86, 57 per cent; p < 0.0001). Twelve (14 per cent) of those not reported had pressure sores and were being cared for on specialist mattresses. Non-reporting was high from all wards. When reports were made of patients at risk of, or with, pressure ulcers, the reports were mostly accurate and complete. CONCLUSION: The data collection form is user-friendly and the assessment skills of nursing staff at the trust are adequate. However, the high incidence of under-reporting indicates that the current system may not be adequate for generating accurate data. PMID- 15132039 TI - Developing a tissue viability nursing assistant role. AB - This article describes the approach taken to introduce and develop the role of the tissue viability nursing assistant in the tissue viability team at the University Hospitals of Leicester NHS Trust. It highlights the achievements and challenges to date and identifies future plans for this role. PMID- 15132040 TI - Prescription drugs uses and effects. Bronchodilators: adrenoreceptor agonists. PMID- 15132041 TI - Hyperglycemic crisis. Regaining control. PMID- 15132042 TI - Patient information. How to prevent high blood sugar. PMID- 15132043 TI - A reawakened loss. PMID- 15132044 TI - Trauma nursing. Traumatic brain injury. PMID- 15132045 TI - Chemo's toll on memory. PMID- 15132046 TI - What's new in the fight against AIDS. PMID- 15132047 TI - Saying the right thing when things go wrong. PMID- 15132048 TI - Keying in on auto clubs. PMID- 15132049 TI - How to shop safely online. PMID- 15132050 TI - Getting "C diff" under control. PMID- 15132051 TI - Tax tips for travelers. PMID- 15132052 TI - Dengue fever, Indonesia--update. PMID- 15132053 TI - Progress towards poliomyelitis eradication in Nigeria, January 2003-March 2004. PMID- 15132054 TI - Nipah virus outbreak(s) in Bangladesh, January-April 2004. PMID- 15132055 TI - Should mentors be paid for the work they do? PMID- 15132056 TI - Opening our doors to new EU nurses. PMID- 15132057 TI - How I coped with ... being struck off. PMID- 15132058 TI - 'We will do everything and anything for our patients'. PMID- 15132059 TI - Increasing awareness of services for people with multiple sclerosis. AB - Awareness of multiple sclerosis has been increasing in recent years. However, there are still many issues which the MS Society will be highlighting during MS Week. This year there are three main MS Week themes. These include funding for research, a volunteer recruitment drive, and promotion of the NICE guideline for MS. PMID- 15132060 TI - What you need to know about ... stroke. PMID- 15132061 TI - The effect of acidic maintenance solutions on catheter longevity. AB - Long-term urinary catheterisation is rarely completely free of complications. Catheter blockage from mineral salt encrustation occurs in 40-50 per cent of patients. Recurrent blockage is distressing to patients and carers and costly to health services in both time and resources. Laboratory-based evidence shows acidic maintenance solutions can be effective in dissolving encrustations. This is supported by clinical experience but there are few clinical studies and no well-controlled clinical trials to date. This article examines the range of evidence for the effect of maintenance solutions on catheter longevity. PMID- 15132063 TI - Planning care for people with a learning disability. PMID- 15132062 TI - Evaluating the funding and capacity of research in Scotland. AB - This paper will discuss the results of a scoping exercise carried out as part of a strategy to inform subsequent models of infrastructure funding. The exercise showed that Scotland has less research capacity in nursing and midwifery than the UK as a whole. Following the publication of a nursing and midwifery research strategy for Scotland, initiatives are being put in place to fund an increase in research capacity and capability in the nursing and midwifery professions. PMID- 15132064 TI - Malignant mesothelioma: risk factors and current management. AB - Malignant mesothelioma is a cancer of the pleura and peritoneum often associated with asbestos exposure. Although rare its incidence is increasing, principally as a result of the long latency period of the disease. This article presents a review of mesothelioma, looking at the disease process, risk factors, causes, and current management strategies--namely surgery, chemotherapy, and radiotherapy. Some of the nursing implications are discussed along with the resources currently available to patients with mesothelioma in the UK. PMID- 15132065 TI - The role of insulin pumps in the management of diabetes. AB - Insulin pumps have been available since the 1980s but despite initial enthusiasm there were problems with pump failure that limited their use. When a pump fails to deliver insulin, diabetic ketoacidosis (DKA) (elevated levels of ketones that occur in response to an imbalance in fat metabolism) can occur. This is potentially fatal for people with type 1 diabetes. The Diabetes Control and Complications Trial (DCCT Research Group, 1993) showed that improved diabetes control reduces diabetic complications. PMID- 15132066 TI - An intensive education programme for people with type 1 diabetes. AB - In the year 2000, approximately 330,000 people in the UK had been diagnosed with type 1 diabetes (Williams and Pickup, 2000), and this number is growing yearly. It is usually diagnosed in children and young adults although it can occur at any age. The onset of diabetes is usually sudden and results from destruction of the insulin-producing cells of the pancreas leading to a total loss of insulin secretion. Insulin injections and dietary modification are necessary treatments after diagnosis. Some patients find these changes difficult--a case study used in this article Illustrates the challenges they face. PMID- 15132067 TI - Perioperative fasting and the management of type 2 diabetes. AB - Type 2 diabetes is the most common form of diabetes, affecting approximately 90 per cent of people with the condition. The global incidence of diabetes is rising and the number of people affected is projected to exceed 300 million by the year 2025 (World Health Organization, 2003). Type 2 diabetes results from insulin resistance and/or a relative lack of insulin production. It is progressive and many patients may require dietary modifications, oral hypoglycaemic medication, and/or insulin to obtain good glycaemic control. Good control has been shown to reduce the complications of diabetes (UK Prospective Diabetes Study Group, 1998). PMID- 15132068 TI - Understanding the processes behind the regulation of blood glucose. AB - Glucose is one of the body's principal fuels. It is an energy-rich monosaccharide sugar that is broken down in our cells to produce adenosine triphosphate. ATP is a small packet of chemical energy that powers the millions of biochemical reactions that take place in the body every second. PMID- 15132069 TI - The Canadian Virtual Hospice . PMID- 15132070 TI - The nature of suffering and its relief in the terminally ill: a qualitative study. AB - The essential mandate of medicine is the relief of suffering. However, the quest for an integrated model towards a conceptualization of suffering is still ongoing and empirical studies are few. Qualitative inquiry using 31 in-depth interviews and content analysis was carried out between 1999 and 2001 in 26 patients diagnosed with terminal cancer. The suffering experience was described through a multiplicity of heterogenous elements from the physical, psychological, and social spheres. Systematic synthesis of interview material yielded three apparently irreducible core dimensions. Respondents defined their suffering in terms of 1) being subjected to violence, 2) being deprived and/or overwhelmed, and 3) living in apprehension. Cassell wrote, in 1991, that to know the suffering of others demands an exhaustive understanding of what makes them the individuals they are (1). Our model can be of use in structuring and eliciting this necessary information. Understanding how a particular patient feels harmed, deprived or overburdened, and overtaken by fear, provides a lever for action tailored to the specifics of that person's experience. PMID- 15132071 TI - How do proxies' perceptions of patients' pain, anxiety, and depression change during the bereavement period? AB - The retrospective approach in palliative care research provides valuable insight into death and dying, and the effectiveness of palliative care. The method involves collecting information from proxies (usually significant others) after the patient's death. This exploratory study investigates whether proxies' accounts differ during bereavement, and provides possible explanations for why discrepancies might occur. Thirteen bereaved family members were interviewed, at three to five months and seven to nine months after the patient's death, about the patient's pain, anxiety, and depression, using semi-structured interviews and the symptom rating scale from the Views of Informal Carers-Evaluation of Services (VOICES) interview. Analysis of VOICES ratings over time indicated consistency for anxiety, while pain and depression ratings were variable and, in many cases, less severe and less frequent with the passage of time. Qualitative analysis of proxies' interview transcripts revealed a number of categories and themes that could be explained within the psychological and palliative care literature. The findings suggest that timing is an important consideration when gathering information from proxies retrospectively. PMID- 15132072 TI - Family perceptions of worry, symptoms, and suffering in the dying. AB - In this exploratory study, family members of 63 decedents were interviewed by telephone, two to five months post-death, about their perceptions of their loved one's worries, symptoms, and suffering at the end of life. The most common worries reported focused on loss of bodily function (44%), being dependent (40%), and being a burden (39%). Distressing physical and psychological symptoms were reported. A majority (94%) of family members reported that their loved one suffered at the end of life, but only worries about loss of quality of life (e.g., being unable to participate in enjoyable activities) were predictive of reports of suffering. Findings suggest that worries, irrespective of the level of current symptoms, play an important role in the suffering of dying patients, and that treatment plans for the terminally ill should routinely explore both symptoms and worries. PMID- 15132073 TI - Esophageal stents in malignant dysphagia: a two-edged sword? AB - Dysphagia from mechanical esophageal obstruction in patients with advanced malignancy is a common and debilitating symptom in patients referred to palliative care services. Relief of such dysphagia is often attempted by insertion of an esophageal stent in the hope that this will improve the quality of life for these patients. We describe a series of 39 patients who had an esophageal stent inserted under radiologic guidance for malignant dysphagia over an 82-month period. While the stents were clearly effective at relieving dysphagia, they also induced significant comorbidity, in particular, moderate to severe chest pain occurring in 46% and reflux esophagitis in 26% of our non selected patient group. In addition we found an eight percent mortality rate from esophageal bleeding following stent insertion. When discussing the potential role of esophageal stent insertion with patients under the care of palliative care teams, the frequency and severity of these significant secondary symptoms need to be considered. PMID- 15132074 TI - The status of undergraduate palliative medicine education in Canada: a 2001 survey. AB - A survey was conducted in fall 2001/spring 2002 to provide an update on the status of undergraduate palliative medicine education in Canada. The survey identified that the majority of palliative care teaching occurs in the pre clinical years of medical school, with supervised patient encounters occurring primarily during electives. The coverage of palliative care topics is inconsistent across curricula. Student evaluation methods also vary, with only one school using simulated patients. More than half the schools have an academic division or department of palliative care medicine, although faculty with protected academic time are few in number. A number of barriers to palliative medicine education were identified, including competition for time within the undergraduate curriculum, and lack of resources for curriculum development and teaching. Respondents recommended increased clinical exposure, curriculum development, student assessment and evaluation, faculty development, and improved infrastructure. Following these recommendations, the Undergraduate Palliative Medicine Committee has dedicated itself to developing and fostering a strategic implementation plan to improve palliative medicine education in Canadian medical schools. PMID- 15132075 TI - Factors associated with caregiver burden among caregivers of terminally ill patients with cancer. AB - OBJECTIVE: To identify factors associated with caregiver burden among those caring for terminally ill patients with cancer. DESIGN: Cross-sectional study of interviews with primary caregivers (n = 206) of consecutive patients with cancer enrolled in the largest hospice in Connecticut. METHODS: Data were collected on the caregivers' sociodemographic characteristics, social network index, and number of restrictions in their own activities due to their caregiving role. The outcome was a nine-item questionnaire adapted from the Zarit Burden Inventory. RESULTS: The highest burden was reported among caregivers with more limited social networks (OR 1.38, Cl 1.02-1.87), more restrictions in their daily activities (OR 1.35, Cl 1.13-1.61), and who were younger (OR 1.46, Cl 1.10-1.93). CONCLUSIONS: Variations exist in the intensity of caregiver burden based on subjective experiences and social support, rather than on the amount of assistance provided. Clinicians should consider factors such as these when targeting caregivers for interventions to alleviate burden. PMID- 15132076 TI - Providing cancer and palliative care in rural areas: a review of patient and carer needs. PMID- 15132077 TI - A systematic review of religion and spirituality in three palliative care journals, 1990-1999. PMID- 15132078 TI - Hospital narratives: "I remember you"--a brief encounter in palliative care. PMID- 15132080 TI - Volunteers are essential members of the clinical team. PMID- 15132079 TI - Maintenance of analgesia with an intrathecal catheter system during an episode of bacterial meningitis. PMID- 15132081 TI - Maxillary midline diastema--aetiology and orthodontic treatment. PMID- 15132082 TI - Bleaching--the facts and the myths. Interview by Aisling O'Mahony. PMID- 15132083 TI - Dentine hypersensitivity: a review of the literature. PMID- 15132084 TI - Teen athletes. PMID- 15132085 TI - Lifestyle activity. Exercise for the masses. PMID- 15132086 TI - Healthy eating. Spring's in the air. Time for a picnic. PMID- 15132087 TI - A drink a day..."Here's to a long life". PMID- 15132088 TI - Beyond the numbers. "...and how do you feel"? PMID- 15132089 TI - 5 things you should know before pumping. If you're ready for one, an insulin pump can change your life for the better. Here, the experts help you prepare for the transition. PMID- 15132090 TI - Roping diabetes. Cowboy Kory Koontz wears his insulin pump into the rodeo arena. PMID- 15132091 TI - Snappy sandwiches. You can have these sandwiches on the table in less than half an hour. PMID- 15132092 TI - Irritable bowel syndrome & diabetes. Having both diabetes and irritable bowel syndrome is frustrating. Here's help controlling both. PMID- 15132093 TI - PCOS. Uncovering polycystic ovary syndrome. PMID- 15132094 TI - Research profile. Hypoglycemia unawareness. Can beta blockers help? Christian Meyer, MD. PMID- 15132095 TI - Help for early Alzheimer's. PMID- 15132096 TI - Surgery in your doctor's office: how safe is it? PMID- 15132097 TI - Your dietary arsenal against 8 serious disorders? PMID- 15132098 TI - Hmmm, did I take that pill...or not? PMID- 15132099 TI - Do sunglasses really protect my eyes--and do darker sunglasses offer the best protection? PMID- 15132100 TI - Grapefruit interacts with many medications. PMID- 15132101 TI - Side air bags save lives. PMID- 15132102 TI - Dry eye syndrome increases with age. PMID- 15132103 TI - Risks of hormone use continue to emerge. Ongoing analyses strengthen case against menopausal hormone therapy. PMID- 15132104 TI - Advances in treating lymphatic cancer. High-dose chemotherapy plus stem-cell support improves survival for some people with non-Hodgkin's lymphoma. PMID- 15132105 TI - Tamoxifen shuffle. PMID- 15132106 TI - Excess wine may promote breast cancer. PMID- 15132107 TI - Antibacterial soap: all washed up? PMID- 15132108 TI - New drug curbs migraine pain. PMID- 15132109 TI - Oral drugs thwart diabetes-driven mental decline. PMID- 15132110 TI - Boosting brittle bones. Fosamax fends off osteoporosis long-term. PMID- 15132112 TI - Feet, don't fail me now! PMID- 15132111 TI - Two RA drugs better than one. PMID- 15132113 TI - Ancient anti-gout diet still works. PMID- 15132114 TI - Tai chi: a bevy of benefits. PMID- 15132115 TI - Exercise activates brain's attention centers. PMID- 15132116 TI - When sinus trouble won't stay away. For people with chronic sinusitis, nasal irrigation and surgery offer avenues for fewer relapses and better breathing. PMID- 15132117 TI - Questioning the power of C-reactive protein to predict heart disease. PMID- 15132118 TI - Heart-attack symptoms vary in older women. PMID- 15132119 TI - No place like home for BP monitoring. PMID- 15132120 TI - Lipitor bests Pravachol at certain doses. PMID- 15132121 TI - Why do people faint? PMID- 15132122 TI - What causes a spermatocele, and how is it usually removed? PMID- 15132123 TI - Management. Out of the rut. PMID- 15132124 TI - US societies to defy ban on editing articles from embargoed countries. PMID- 15132125 TI - Has the Presidents's Council on Bioethics missed the boat? PMID- 15132126 TI - Clinical and attentional effects of acute nicotine treatment in Tourette's syndrome. AB - Evidence from pre-clinical infrahuman investigations, open-label clinical trials, and a single controlled trial found acute nicotine treatment potentiated up to 4 weeks neuroleptic-induced reductions of dyskinetic symptoms characterizing Tourette's syndrome (TS). Given the attentional disturbances associated with this syndrome, and the improvements in attentional processes reported with nicotine, this randomized, double-blind, placebo-controlled trial examined the acute (4 h) and sustained (2 weeks) effects of a single dose of transdermal nicotine on clinical (i.e., tics), attentional (continuous performance task, event-related potential, patient and parental reports) and behavioral symptoms in 23 children and adolescents with TS receiving neuroleptic treatment. In the 14 evaluable patients with complete primary efficacy data, nicotine (compared to placebo) failed to alter symptoms at 4 h but counteracted ERP-P300 signs of diminished attention seen 2 weeks following placebo treatment. Secondary efficacy measures, including patient self-reports and parental ratings, found nicotine to reduce complex tics and improve behaviors related to inattention. Additional work with intermittent dosing schedules is required to characterize optimal clinical and cognitive effects with nicotine treatment. PMID- 15132127 TI - Current awareness on yeast. PMID- 15132128 TI - Enzyme expression profiles suggest the novel tumor-activated fluoropyrimidine carbamate capecitabine (Xeloda) might be effective against papillary thyroid cancers of children and young adults. AB - PURPOSE: The fluoropyrimidine carbamate (capecitabine) is converted to 5 fluorouracil (5-FU) by thymidine phosphorylase (TP) inside target tissues. 5-FU interferes with DNA synthesis by blocking thymidylate synthase (TS) but is inactivated by dihydropyrimidine dehydrogenase (DPD). Favorable enzyme profiles (high TP and low DPD) generate high intratumor levels of 5-FU that are effective against many tumors, especially those with low TS. Capecitabine has not been tested against thyroid cancers, and it is not known to what extent thyroid cancers express TP, TS or DPD. METHODS: To test this, we determined TP, TS and DPD in 19 thyroid cancers from young patients (14 papillary, 4 follicular, 1 medullary) by immunohistochemistry. After approval by the Human Use Committee, the intensity of TP, TS, and DPD staining was determined by two independent examiners and graded (absent=0 to intense=3) with >90% concordance. RESULTS: TS was detected in 7/19 cancers (37%), TP in 14/19 cancers (74%) and DPD in 14/19 cancers (74%). In six tumors, TP was more intense that DPD, suggesting capecitabine sensitivity. Only five tumors failed to express TP but four of these expressed DPD, suggesting capecitabine resistance. Overall, 6/19 tumors (32% of the total) had a favorable expression profile, and all of them were papillary cancers. CONCLUSIONS: We conclude that the majority of differentiated thyroid cancers (74%) express TP and low levels of TS (63% undetectable). The results support the hypothesis that capecitabine is activated in the majority of differentiated thyroid cancers and that 32% have favorable expression of all three enzymes (TP, TS, and DPD). PMID- 15132129 TI - Comparison of intraperitoneal continuous infusion of floxuridine and bolus administration in a peritoneal gastric cancer xenograft model. AB - PURPOSE: To identify the optimal schedule for intraperitoneal (i.p.) infusion of floxuridine (FUDR) against peritoneal micrometastases from gastric cancer. METHODS: The efficacy of continuous i.p. infusion of FUDR was compared with that of bolus i.p. administration in peritoneal gastric cancer (MKN45) xenografts. The FUDR continuous delivery system in this study was in the form of injectable poly(lactic-coglycolic) acid (PLGA) microspheres intended for i.p. injection. Animals were treated by continuous i.p. infusion using FUDR-loaded microspheres or bolus i.p. administration of FUDR. RESULTS: In vitro testing demonstrated that FUDR was released slowly from the microspheres at a rate of approximately 5% of the total encapsulated drug per day. In in vivo studies, the peritoneal level was found to persist and was approximately 5- to 50-fold higher than that of plasma for more than 2 weeks following a single injection of the microspheres. An in vitro MTT assay showed that exposure time clearly influenced the cytotoxic potency of FUDR. In vivo, continuous infusion was more effective against peritoneal tumor than bolus administration at equivalent doses. However, compared with bolus administration, toxicity was increased, resulting in a reduced maximum tolerated dose (MTD) with continuous infusion. When the treatment was carried out at each MTD (continuous 1 mg/kg, bolus 600 mg/kg), continuous infusion had no advantage in inhibiting tumor growth. CONCLUSIONS: Owing to the higher toxicity and the equal efficacy of continuous infusion compared with bolus administration, continuous infusion is not recommended in i.p. FUDR treatment. PMID- 15132130 TI - Antimalarial dihydroartemisinin also inhibits angiogenesis. AB - Dihydroartemisinin, a more water-soluble metabolite of artemisinin derivatives, is a safe and most effective antimalarial analog of artemisinin. In the present study, we investigated the antiangiogenic activity of dihydroartemisinin in vitro and in vivo, and investigated dihydroartemisinin-induced apoptosis in human umbilical vein endothelial cells (HUVEC). Dihydroartemisinin markedly reduced VEGF binding to its receptors on the surface of HUVEC. The expression levels of two major VEGF receptors, Flt-1 and KDR/flk-1, on HUVEC were lower following dihydroartemisinin treatment as shown by an immunocytochemical staining assay. The in vivo antiangiogenic activity was evaluated in the model of chicken chorioallantoic membrane (CAM) neovascularization. Dihydroartemisinin significantly inhibited CAM angiogenesis at low concentrations (5-30 nmol/100 microl per egg). We also investigated both qualitatively and quantitatively the induction of HUVEC apoptosis by dihydroartemisinin. A dose-related (5-80 microM) and time-dependent (6-36 h) increase in dihydroartemisinin-induced HUVEC apoptosis was observed by flow cytometry. Our results suggest that the antiangiogenic effect induced by dihydroartemisinin might occur by induction of cellular apoptosis and inhibition of expression of VEGF receptors. These findings and the known low toxicity of dihydroartemisinin indicate that it might be a promising candidate angiogenesis inhibitor. PMID- 15132131 TI - Bioequivalence, safety, and tolerability of imatinib tablets compared with capsules. AB - PURPOSE: Imatinib (Glivec) has been established as a highly effective therapy for chronic myeloid leukemia and gastrointestinal tumors. The recommended daily dosage of 400-600 mg requires simultaneous intake of up to six of the current 100 mg capsules. Due to the need to swallow multiple capsules per dose, there is a potential negative impact on treatment adherence; therefore, a new imatinib 400 mg film-coated tablet has been developed. To improve dosing flexibility, particularly with regard to the pediatric population and the management of adverse events, a scored 100-mg film-coated tablet has also been introduced. EXPERIMENTAL DESIGN: A group of 33 healthy subjects were randomly assigned to one of six treatment sequences, in which they received imatinib as 4 x 100-mg capsules (reference), 4 x 100-mg scored tablets (test), and 1 x 400-mg tablet (test). Blood sampling was performed for up to 96 h after dosing, followed by a 10-day washout period prior to the next sequence. After the third dosing, subjects were monitored to assess delayed drug-related adverse events. Pharmacokinetic parameters were assessed using concentration-time curves for plasma imatinib and its metabolite CGP74588. RESULTS: Median Tmax was 2.5 h for capsules and tablets. Mean AUC((0-inf)) values were 27,094, 26,081 and 25,464 ng.h/ml for 4 x 100-mg capsules, 4 x 100-mg tablets, and 1 x 400-mg tablets, respectively. Cmax values were 1748, 1638 and 1606 ng/ml, and t(1/2) values were 15.8, 15.9 and 15.7 h. The test/reference ratios for AUC((0-inf)), AUC((0-96) (h)), and C(max) were 0.98, 0.98 and 0.95 for 4 x 100-mg tablets versus 4 x 100 mg capsules, and 0.95, 0.95 and 0.92 for 1 x 400-mg tablet versus 4 x 100-mg capsules. The 95% confidence intervals were fully contained within the interval (0.80, 1.25). Eight mild and one moderate adverse event considered to be drug related were reported. These events showed no clustering by type of dosage form and were of little to no clinical significance. CONCLUSIONS: Film-coated 100-mg (scored) and 400-mg tablet dose forms of imatinib are bioequivalent to the commercial 100-mg hard-gelatin capsule, and are as safe and well tolerated. PMID- 15132132 TI - Naturally occurring variants in catecholamine receptor genes. PMID- 15132133 TI - Alpha2-adrenoceptor subtypes--surprising insights from gene-targeted mouse models. PMID- 15132134 TI - NO enhances noradrenaline release: modulation and mechanism. PMID- 15132135 TI - Carboplatin and docetaxel in advanced non-small-cell lung cancer: results of a multicenter phase II study. AB - BACKGROUND: To evaluate the efficacy of carboplatin and docetaxel combination in patients with advanced non-small-cell lung cancer. METHODS: In a phase II study, patients with inoperable stage IIIB or stage IV non-small-cell lung cancer (ECOG performance status of 0 or 1) were treated with the combination of carboplatin AUC 5 mg/ml.min and docetaxel 80 mg/m2 administered once every 3 weeks. RESULTS: A total of 45 patients were accrued to the study. The median age was 62 years and adenocarcinoma was the most common histology. Patients received a median of four cycles of chemotherapy. The objective response rate was 29% with a median survival of 11.9 months among evaluable patients. The 1-year survival rate was 47%. Febrile neutropenia (17%) was the most common hematological toxicity associated with the regimen whereas grade 3 fatigue (4%) was the major nonhematological toxicity. CONCLUSIONS: The combination of carboplatin plus docetaxel is well tolerated and is effective for the treatment of patients with previously untreated advanced or metastatic non-small-cell lung cancer. PMID- 15132136 TI - Dihydropyrimidine dehydrogenase pharmacogenetics in the Taiwanese population. AB - BACKGROUND/PURPOSE: 5-Fluorouracil (5-FU) remains the most frequently used chemotherapy agent in various human cancers. Over 80% of the 5-FU administered is metabolized by dihydropyrimidine dehydrogenase (DPD) in the liver. However, mutations in the DPD gene have been found to be associated with low DPD activity causing severe complications. The aim of this study was to determine the frequency of 11 known mutations in Taiwanese subjects and the relationship between mutation and DPD level. METHODS: Samples from a total of 300 subjects were investigated in this study. The PCR-RFLP method was used to identify 11 mutations of the DPYD gene, including 62G>A, 74A>G, 85T>C (DPYD*9A), 812delT, 1003G>T, 1156G>T, 1627A>G (DPYD*5), 1714C>G, 1897delC (DPYD*3), 2194G>A (DPYD*6), and IVS14+1G>A (DPYD*2A). DPD protein levels were determined using a DPD ELISA kit. RESULTS: Four mutations, including 74A>G, 85T>C (DPYD*9A), 1627A>G (DPYD*5), and 2194G>A (DPYD*6), were found in our 300 samples. The following mutations were not detected: 62G>A, 812delT, 1003G>T, 1156G>T, 1714C>G, 1897delC (DPYD*3), and IVS14+1G>A (DPYD*2A). The phenotype analysis by DPD protein level indicated that the 1627A>G (DPYD*5) mutation was not associated with the DPD protein level and might be a polymorphism in the DPD gene. The DPD level was also not correlated with gender. CONCLUSION: No significant correlations between these 11 mutations and DPD protein level were found indicating that examination of these mutations is insufficient to provide a high-value prediction of the 5-FU pharmacogenetic syndrome in Taiwanese. Genotype and phenotype analysis indicated the 1627A>G (DPYD*5) mutation to be a polymorphism. PMID- 15132137 TI - US academics forbidden to attend Cuban coma conference. PMID- 15132138 TI - Out with the old and in with the new...epilepsy drugs. PMID- 15132139 TI - Heart association urges passage of US stroke legislation. PMID- 15132140 TI - Three journals raise doubts on validity of Canadian studies. PMID- 15132141 TI - Computerised system translates "subvocal speech" into action. PMID- 15132142 TI - Fall of a dictator, failure of ethics. PMID- 15132143 TI - Exodus of Czech doctors leaves gaps in health care. Hospital officials draw up crisis plans to prepare for May 1 easing of migration restrictions. PMID- 15132144 TI - [Conformal index and radiotherapy]. AB - Goal of radiotherapy is to treat patient with the best therapeutic ratio, i.e. the highest local control and the lowest toxicity rates. The conformal approach, three-dimensional conformal radiotherapy or intensity-modulated radiotherapy, is based on imageries, up-dated 3-D treatment planning systems, immobilization systems, restricted quality assurance and treatment verification. The aim is to ensure a high dose distribution tailored to the limits of the target volume, while reducing exposure of normal tissues. The evaluation tools used for optimizing treatment are the visual inspection of the dose distribution in various planes, and the dose-volume histograms, but they do not fully quantify the conformity of dose distributions. The conformal index is a tool for scoring a given plan or for evaluating different treatment plans for the same patient. This paper describes the onset and evolution of conformal index and his potential application field. PMID- 15132145 TI - [Image processing and radiotherapy]. AB - Medical images are of great importance in radiotherapy, which became a privileged application field for image processing techniques. Moreover, because of the progression of the computers' performances, these techniques are also in full expansion. Today, the recent developments of the radiotherapy (3DCR, IMRT) offer a huge place to them. Effectively, they can potentially answer to the precision requirements of the modern radiotherapy, and may then contribute to improve the delivered treatments. The purpose of this article is to present the different image processing techniques that are currently used in radiotherapy (including image matching and segmentation) as they are described in the literature. PMID- 15132146 TI - Medicare program; prospective payment system for long-term care hospitals: annual payment rate updates and policy changes. Final rule. AB - This final rule updates the annual payment rates for the Medicare prospective payment system (PPS) for inpatient hospital services provided by long-term care hospitals (LTCHs). The payment amounts and factors used to determine the updated Federal rates that are described in this final rule have been determined based on the LTCH PPS rate year. The annual update of the long-term care diagnosis-related group (LTC-DRG) classifications and relative weights remains linked to the annual adjustments of the acute care hospital inpatient diagnosis-related group system, and will continue to be effective each October 1. The outlier threshold for July 1, 2004 through June 30, 2005 is also derived from the LTCH PPS rate year calculations. In this final rule, we also are making clarifications to the existing policy regarding the designation of a satellite of a LTCH as an independent LTCH. In addition, we are expanding the existing interrupted stay policy and changing the procedure for counting days in the average length of stay calculation for Medicare patients for hospitals qualifying as LTCHs. PMID- 15132147 TI - Medicare program; changes to the criteria for being classified as an inpatient rehabilitation facility. Final rule. AB - This final rule responds to public comments on the September 9, 2003 proposed rule, and revises the classification criterion, commonly known as the "75 percent rule," used to classify a hospital as an inpatient rehabilitation facility (IRF). This final rule also modifies and expands the medical conditions listed in the regulatory requirements as well as temporarily lowers the percentage of patients required to fall within one of the specified list of medical conditions. PMID- 15132148 TI - Explosion in North Korea destroys health facilities. PMID- 15132149 TI - WHO uses Malaria Day to promote ACTs. PMID- 15132150 TI - Manufacturers face jail after baby milk kills 50 in China. PMID- 15132151 TI - More aid is needed to halve world poverty, says report. PMID- 15132152 TI - UK health reforms get off to a shaky start. Foundation hospitals suffer publicity blows and private partnership plans hit a stumbling block. PMID- 15132153 TI - Giant uterine leiomyoma with air-fluid levels. PMID- 15132154 TI - Prostacyclin/thromboxane early changes in pregnancies that are complicated by preeclampsia. PMID- 15132155 TI - Duration of pregnancy and sex of offspring. PMID- 15132156 TI - Long-term neurodevelopmental outcome after intrauterine laser treatment for twin twin transfusion syndrome (TTTS). PMID- 15132157 TI - Twins conceived through in vitro fertilization. PMID- 15132158 TI - Soda isn't only low in calcium. PMID- 15132159 TI - Soda isn't only low in calcium. PMID- 15132162 TI - Toward better understanding of equivocal deaths and suicides. PMID- 15132163 TI - Role of journals in addressing scientific misconduct. PMID- 15132161 TI - Late onset of motor neurons in mice overexpressing wild-type peripherin. AB - Peripherin, a type III intermediate filament (IF) protein, upregulated by injury and inflammatory cytokines, is a component of IF inclusion bodies associated with degenerating motor neurons in sporadic amyotrophic lateral sclerosis (ALS). We report here that sustained overexpression of wild-type peripherin in mice provokes massive and selective degeneration of motor axons during aging. Remarkably, the onset of peripherin-mediated disease was precipitated by a deficiency of neurofilament light (NF-L) protein, a phenomenon associated with sporadic ALS. In NF-L null mice, the overexpression of peripherin led to early- onset formation of IF inclusions and to the selective death of spinal motor neurons at 6 mo of age. We also report the formation of similar peripherin inclusions in presymptomatic transgenic mice expressing a mutant form of superoxide dismutase linked to ALS. Taken together, these results suggest that IF inclusions containing peripherin may play a contributory role in motor neuron disease. PMID- 15132164 TI - [Systemic lupus erythematosus]. PMID- 15132165 TI - Structure of N-glycosidic carbohydrates of secretory proteins of Tetrahymena thermophila. AB - Glycoproteins secreted by Tetrahymena into the culture medium were isolated and the N-glycosidic oligosaccharides analyzed using lectin blots and fluorophore assisted carbohydrate gel electrophoresis (FACE). Lectin blots showed that the glycoproteins secreted by Tetrahymena contain only N-glycosidic structures of the high mannose type. Further analysis using the FACE technology revealed the presence of four different N-glycosidic structures differing only in the number of mannose residues attached to the core chitobiose unit. PMID- 15132166 TI - Ultrastructure of Besnoitia besnoiti tissue cysts and bradyzoites. AB - Besnoitia besnoiti is an economically important tissue cyst-forming apicomplexan of cattle in Africa and Israel. Tissue cysts and bradyzoites of B. besnoiti from the skin of a naturally infected bull were studied by transmission electron microscopy. Tissue cysts enclosed host cell and bradyzoites. Bradyzoites were 6 7.5 x 1.9-2.3 microm in size and contained organelles found in coccidian merozoites including numerous micronemes, rhoptries, amylopectin granules, and a posteriorly located nucleus. Enigmatic bodies, characteristically found in Besnoitia sp. bradyzoites, were not observed. Therefore, enigmatic bodies should be removed as a generic character of the bradyzoites of Besnoitia species. PMID- 15132167 TI - Locomotion and phenotypic transformation of the amoeboflagellate Naegleria gruberi at the water-air interface. AB - The protozoon Naegleria gruberi is able to carry out amoeboid locomotion at the water-air interface in a manner indistinguishable from that exhibited on solid substrata with the production of focal contacts and associated filopodia. The speed of locomotion at this interface can be modulated by changes in electrolyte concentrations; these speed changes are identical to those observed at a water glass interface. The nature of the water-air interface is discussed leading to the hypothesis that surface tension alone could provide suitable properties for the adhesion and translocation of amoebae at this interface without necessitating specific, absorbed molecules. The temporary swimming flagellate stage of Naegleria is able to dock at the interface, make stable adhesions to it, and revert to the amoeboid phenotype. Conversely, amoebae resident at the water-air interface can transform to swimming flagellates and escape into the bulk liquid phase. We report the presence of Naegleria amoebae in the surface microlayers of natural ponds; thus, in freshwater bodies there may be active shuttling of Naegleria amoebae from the benthos to the surface microlayers by means of the non feeding, swimming flagellate phenotype. The public health implication of this behaviour in the case of the pathogenic relative, Naegleria fowleri, is discussed. PMID- 15132168 TI - Profilin functions in cytokinesis, nuclear positioning, and stomatogenesis in Tetrahymena thermophila. AB - Expression of the actin-binding protein profilin was disrupted in the ciliate Tetrahymena thermophila by an antisense ribosome method. In cells with the antisense disruption no profilin protein was detected. Cultures of cells with the antisense disruption could be maintained, indicating that profilin was not essential for cytokinesis or vegetative growth. Disruption of the expression of profilin resulted in many cells that were large and abnormally shaped. Formation of multiple micronuclei, which divide mitotically, was observed in cells with a single macronucleus, indicating a defect in early cytokinesis. Some cells with the antisense disruption contained multiple macronuclei, which in Tetrahymena may indicate a function late in cytokinesis. The lack of profilin also affected cytokinesis in the cells that could divide. Normal-sized and normal-shaped cells with the antisense disruption took significantly longer to divide than control cell types. The profilin disruption revealed two new processes in which profilin functions. In cells lacking profilin, micronuclei were not positioned at their normal site on the surface of the macronucleus and phagocytosis was defective. The defect in phagocytosis appeared to be due to disruption of the formation of oral apparatuses (stomatogenesis) and a possible failure in the internalization of phagocytic vacuoles. PMID- 15132169 TI - Detection of Naegleria sp. in a thermal, acidic stream in Yellowstone National Park. AB - An initial survey of sequences of PCR-amplified portions of the 18S rRNA genes from a community DNA clone library, prepared from an algal mat in a thermal, acidic stream in Yellowstone National Park, WY, USA, revealed among other sequences, several that matched Vahlkampfia. This finding prompted further investigation using primers specific for Naegleria. Sequences from a subsequent DNA clone library, prepared from the 5.8S rRNA gene and the adjacent internal transcribed spacer (ITS) regions of the rRNA, closely matched Naegleria and formed an independent lineage within a clade containing Naegleria sturti and Naegleria niuginiensis. The sequences may represent a new Naegleria species. PMID- 15132170 TI - Resolution of a taxonomic conundrum: an ultrastructural and molecular description of the life cycle of Pleistophora mulleri (Pfeiffer 1895; Georgevitch 1929). AB - The classification of a microsporidian parasite observed in the abdominal muscles of amphipod hosts has been repeatedly revised but still remains inconclusive. This parasite has variable spore numbers within a sporophorous vesicle and has been assigned to the genera Glugea, Pleistophora, Stempellia, and Thelohania. We used electron microscopy and molecular evidence to resolve the previous taxonomic confusion and confirm its identification as Pleistophora mulleri. The life cycle of P. mulleri is described from the freshwater amphipod host Gammarus duebeni celticus. Infection appeared as white tubular masses within the abdominal muscle of the host. Light and transmission electron microscope examination revealed the presence of an active microsporidian infection that was diffuse within the muscle block with no evidence of xenoma formation. Paucinucleate merogonial plasmodia were surrounded by an amorphous coat immediately external to the plasmalemma. The amorphous coat developed into a merontogenetic sporophorous vesicle that was present throughout sporulation. Sporogony was polysporous resulting in uninucleate spores, with a bipartite polaroplast, an anisofilar polar filament and a large posterior vacuole. SSU rDNA analysis supported the ultrastructural evidence clearly placing this parasite within the genus Pleistophora. This paper indicates that Pleistophora species are not restricted to vertebrate hosts. PMID- 15132171 TI - Feeding by the heterotrophic dinoflagellate Oxyrrhis marina on the red-tide raphidophyte Heterosigma akashiwo: a potential biological method to control red tides using mass-cultured grazers. AB - As part of the development of a method to control the outbreak and persistence of red tides using mass-cultured heterotrophic protist grazers, we measured the growth and ingestion rates of cultured Oxyrrhis marina (a heterotrophic dinoflagellate) on cultured Heterosigma akashiwo (a raphidophyte) in bottles in the laboratory and in mesocosms (ca. 60 liter) in nature, and those of the cultured grazer on natural populations of the red-tide organism in mesocosms set up in nature. In the bottle incubation, specific growth rates of O. marina increased rapidly with increasing concentration of cultured prey up to ca. 950 ng C ml(-1) (equivalent to 9,500 cells ml(-1)), but were saturated at higher concentrations. Maximum specific growth rate (mumax), KGR (prey concentration sustaining 0.5 mumax) and threshold prey concentration of O. marina on H. akashiwo were 1.43 d(-1), 104 ng C ml(-1), and 8.0 ng C ml(-1), respectively. Maximum ingestion and clearance rates of O. marina were 1.27 ng C grazer(-1) d( 1) and 0.3 microl grazer(-1) h(-1), respectively. Cultured O. marina grew well effectively reducing cultured and natural populations of H. akashiwo down to a very low concentration within 3 d in the mesocosms. The growth and ingestion rates of cultured O. marina on natural populations of H. akashiwo in the mesocosms were 39% and 40%, respectively, of those calculated based on the results from the bottle incubation in the laboratory, while growth and ingestion rates of cultured O. marina on cultured H. akashiwo in the mesocosms were 55% and 36%, respectively. Calculated grazing impact by O. marina on natural populations of H. akashiwo suggests that O. marina cultured on a large scale could be used for controlling red tides by H. akashiwo near aquaculture farms that are located in small ponds, lagoons, semi-enclosed bays, and large land-aqua tanks to which fresh seawater should be frequently supplied. PMID- 15132172 TI - The molecular evolution and structural organization of self-splicing group I introns at position 516 in nuclear SSU rDNA of myxomycetes. AB - Group I introns are relatively common within nuclear ribosomal DNA of eukaryotic microorganisms, especially in myxomycetes. Introns at position S516 in the small subunit ribosomal RNA gene are particularly common, but have a sporadic occurrence in myxomycetes. Fuligo septica, Badhamia gracilis, and Physarum flavicomum, all members of the family Physaraceae, contain related group IC1 introns at this site. The F. septica intron was studied at the molecular level and found to self-splice as naked RNA and to generate full-length intron RNA circles during incubation. Group I introns at position S516 appear to have a particularly widespread distribution among protists and fungi. Secondary structural analysis of more than 140 S516 group I introns available in the database revealed five different types of organization, including IC1 introns with and without His-Cys homing endonuclease genes, complex twin-ribozyme introns, IE introns, and degenerate group I-like introns. Both intron structural and phylogenetic analyses indicate a multiple origin of the S516 introns during evolution. The myxomycete introns are related to S516 introns in the more distantly related brown algae and Acanthamoeba species. Possible mechanisms of intron transfer both at the RNA- and DNA-levels are discussed in order to explain the observed widespread, but scattered, phylogenetic distribution. PMID- 15132173 TI - The endosymbiotic bacterium Holospora obtusa enhances heat-shock gene expression of the host Paramecium caudatum. AB - The bacterium Holospora obtusa is a macronuclear-specific symbiont of the ciliate Paramecium caudatum. H. obtusa-bearing paramecia could survive even after the cells were quickly heated from 25 degrees C to 35 degrees C. To determine whether infection with H. obtusa confers heat shock resistance on its host, we isolated genes homologous to the heat shock protein genes hsp60 and hsp70 from P. caudatum. The deduced amino acid sequences of both cDNAs were highly homologous to hsp family sequences from other eukaryotes. Competitive PCR showed that H. obtusa-free paramecia expressed only trace amounts of hsp60 and hsp70 mRNA at 25 degrees C, but that expression of hsp70 was enhanced immediately after the cells were transferred to 35 degrees C. H. obtusa-bearing paramecia expressed high levels of hsp7O mRNA even at 25 degrees C and the level was further enhanced when the cells were incubated at 35 degrees C. In contrast, the expression pattern of hsp60 mRNA was the same in H. obtusa-bearing as in H. obtusa-free paramecia. These results indicate that infection with its endosymbiont can confer a heat shock resistant nature on its host cells. PMID- 15132174 TI - Catalase is the bacteria-derived detoxifying substance against paramecia-killing toxin in wheat grass powder infusion. AB - Paramecium cells are usually cultured in a wheat grass powder infusion inoculated with Klebsiella pneumoniae. However, non-bacterized wheat grass powder infusion is toxic to paramecia, and bacteria-derived substance detoxifies the toxic substance. Here, the detoxifying substance from K. pneumoniae, which was found to be proteinaceous, was purified to homogeneity. The protein had an apparent molecular mass of about 200 kDa by gel filtration and 92 kDa by SDS polyacrylamide gel electrophoresis. Although the amino acid sequence of the amino terminal region did not show a high sequence homology with any reported proteins, amino acid sequences of internal regions of the protein were nearly identical to catalase HPII from Escherichia coli. When the wheat grass powder infusion was treated at 25 degrees C for 1 h with commercially available catalase from bovine liver, the toxicity of the infusion against paramecia was completely abolished. The initial concentration of hydrogen peroxide in the wheat grass powder infusion was about 30 microM and was completely decomposed by the catalase treatment. Therefore, the toxic substance in the wheat grass powder infusion and the detoxifying substance from K. pneumoniae are considered as hydrogen peroxide and catalase, respectively. PMID- 15132175 TI - Aqueous preparation and evaluation of albumin-chitosan microspheres containing indomethacin. AB - Controlled-release egg albumin-chitosan microspheres containing indomethacin as a model drug were successfully prepared by coacervation method. The proposed method can offer a simple method for microsphere preparation in an aqueous system with the elimination of the use of organic solvents that are usually needed in conventional techniques of microencapsulation. The interaction between negatively charged egg albumin molecules in phosphate buffer, pH 7.2, or sodium hydroxide solution and positively charged chitosan molecules dissolved in diluted acetic acid to form an insoluble precipitate was the principle for the formation of the microspheres. The effects of many process variables, such as amount of formaldehyde as a cross-linking agent, stirring time, final pH of encapsulation medium, initial drug loading, and albumin concentration or albumin-to-chitosan weight ratio, on the properties of the prepared microspheres were investigated. Incorporation efficiencies of the microspheres to the drug were high in most cases and ranged between 63.3 +/- 3.6% and 92.39 +/- 3.2%, while particle sizes were 435.2 +/- 12.6 up to 693.9 +/- 34.6 microm for the different tested batches. On the other hand, the values of angles of repose and compressibility indices were in the range of 23.5 +/- 0.4 to 32.0 +/- 0.7 degrees and 11.1 +/- 0.7% to 23.6 +/- 0.7% respectively, which indicate overall good free flowing nature of the microspheres of all batches. The maximum required amount of the cross-linking agent was determined to avoid excessive unnecessary chemicals. It was also noticed that excessive time of stirring and excessive initial drug loading are not recommended as it may lead to microspheres of low properties. The pH of the encapsulation media (pH 3.77 up to pH 4.91) significantly affected the properties of the microspheres. As the pH of the encapsulation media was increased, the incorporation efficiency, particle size, and flowability decreased, along with increase of drug release rate, which could be related to incomplete cross linking of the microspheres matrix. It was also observed that high concentration of albumin solution and accordingly the increase of albumin-to-chitosan weight ratio were accompanied with increases in incorporation efficiency and particle size with improved microsphere flowability and slow indomethacin release. Thus, the proposed microspheres showed the ability to control the release of indomethacin, and their properties were highly affected by many process variables that could be controlled to obtain an optimized system. PMID- 15132176 TI - Effect of postmolding heat treatment on in vitro properties of a polyanhydride implant containing gentamicin sulfate. AB - A polyanhydride implant containing gentamicin sulfate was fabricated using a laboratory-scale injection-molding machine. After injection molding, the implants were subject to heat treatment at 60 degrees C for various time periods with or without nitrogen protection. The impact of this heat treatment on the in vitro properties of the implants including copolymer molecular weights, mechanical properties, and in vitro drug-release profiles was investigated. This heat treatment caused a drastic drop in the molecular weight of the copolymer. Heating without nitrogen protection resulted in the hardening of the implant, but heating in the presence of nitrogen rendered the implant less rigid. It was also found that a faster in vitro drug release profile was shown by implants heated without nitrogen protection and a pronounced slowing down in drug release was exhibited by implants heated with nitrogen protection. PMID- 15132177 TI - Development of local injectable dental gel: the influence of certain additives on physicochemical properties of glycerylmonooleate-based formulations. AB - The current research study is based on the design and development of a sol-gel biodegradable controlled-release formulation for use in the treatment of periodontal diseases. Glycerylmonooleate (GMO) was used as a main composition in the gel base. The influence of various additives, e.g., glycerylmonostearate (GMS), methylcellulose (MC), surfactants, and triglycerides, in GMO formulations on rheologic and swelling properties and release characteristics was described. It was demonstrated that the surfactants and triglycerides affected rheologic behavior, whereas GMS and MC influenced both rheologic and swelling properties of the bases. The release study revealed that drug released from the gel bases depended on the square root of time. The kinetics can be explained by the Higuchi's diffusion theory. Some polyols could enhance drug release from the gel. The stability results suggested that the dental gels obtained should be kept in the low temperature range. PMID- 15132178 TI - Insulin-loaded calcium pectinate nanoparticles: effects of pectin molecular weight and formulation pH. AB - Insulin-loaded calcium pectinate nanoparticles were prepared as a potential colonic delivery system by ionotropic gelation with calcium ions. The effects of pectin molecular weight (Mv) and formulation pH on the characteristics of the nanoparticles were evaluated. Commercial pectins, LM101 and LM104, with respective degrees of esterification of 36% and 28%, were depolymerized by mechanical milling to give Mv ranging from 89 to 5.6 kDa. Milled pectins did not yield nanoparticles with significantly different mean diameter and insulin association efficiency (AE) compared to nanoparticles of unmilled pectins. LM104 nanoparticles had smaller variation in mean size than the LM101 nanoparticles. Formulation pH significantly influenced the AE and stability of the nanoparticles. Increasing the pH from 2 to 3 enhanced the AE by three-fold, from 32.76% to 93.31%, at an insulin loading concentration of 80 U/mL. This increase in AE was correlated to the charge density on the pectin molecules as a function of pH. Subsequent release of associated insulin from the nanoparticles was dependent on the extent of dilution of the nanoparticle dispersion and the pH of the dissolution medium. Cross-flow filtration could be used to separate the nanoparticles from unassociated ions and molecules, without compromising the characteristics of the nanoparticles. PMID- 15132179 TI - Optimization of biodegradable sponges as controlled release drug matrices. I. Effect of moisture level on chitosan sponge mechanical properties. AB - Cross-linked chitosan sponges as controlled release drug carrier systems were developed. Tramadol hydrochloride, a centrally acting analgesic, was used as a model drug. The sponges were prepared by freeze-drying 1.25% and 2.5% (w/w) high and low M.wt. chitosan solutions, respectively, using glutaraldehyde as a cross linking agent. The hardness of the prepared sponges was a function of glutaraldehyde concentration and volume where the optimum concentration that offered accepted sponge consistency was 5%. Below or above 5%, very soft or very hard and brittle sponges were obtained, respectively. The determined drug content in the prepared sponges was uniform and did not deviate markedly from the calculated amount. Scanning electron microscopy (SEM) was used to characterize the internal structures of the sponges. The SEM photos revealed that cross-linked high M.wt. chitosan sponges have larger size surface pores that form connections (channels) with the interior of the sponge than cross-linked low M.wt. ones. Moreover, crystals of the incorporated Tramadol hydrochloride were detected on the lamellae and within pores in both chitosan sponges. Differences in pore size and dissolution medium uptake capacity were crucial factors for the more delayed drug release from cross-linked low M.wt. chitosan sponges over high M.wt. ones at pH 7.4. Kinetic analysis of the release data using linear regression followed the Higuchi diffusion model over 12 hours. Setting storage conditions at room temperature under 80-92% relative humidity resulted in soft, elastic, and compressible sponges. PMID- 15132180 TI - Surface treatment of indomethacin agglomerates with eudragit. AB - Indomethacin is a widely used anti-inflammatory drug with serious side-effects. This drug was used as a model drug for the coating of agglomerates with a permeable film (Eudragit NE). The agglomeration of the crystals increased the flowability of the bulk crystals. The coating further improved the flowability, and also the uniformity of the mass of the filled capsules. The coating film also influenced the wetting of the samples. The coating decreased the surface free energy and therefore reduced the adhesion forces between both the dry and the wet particles. The modification of the flow properties and the even capsule filling can be explained by this phenomenon. Since coating film does not dissolve in the artificial gastric juice, the dissolution test was performed only in the artificial intestinal juice. The dissolution of indomethacin from the coated sample was changed significantly. Accordingly, coating of the crystals can be performed in order to protect the mucosa of the gastrointestinal tract or to promote the preparation of solid dosage form. PMID- 15132181 TI - Stability indicating method for SPf66 antimalarial peptide in solution. AB - Stability studies on the SPf66 antimalarial peptide with different pH and temperature conditions were carried out. The degradation mechanism was elucidated by the size-exclusion chromatography (SEC) technique and the experimental data obtained at 37 degrees C and different pH were fitted to a kinetic degradation model that could explain the loss of its immunogenic capacity. At 5, 25, 37, and 70 degrees C and pH 2, changes were detected in the areas of the different species, although the values obtained could not be fitted to any known degradation kinetics. PMID- 15132182 TI - Transdermal delivery controlled by a chitosan membrane. AB - The release of a drug from a transdermal delivery system with a rate controlling chitosan membrane was analyzed in vitro and in vivo. Lidocaine hydrochloride, a local anesthetic, was used as the model drug. The in vitro permeability of various chitosan membranes for the drug was investigated using a Franz diffusion cell. Drug release was slower through chitosan membranes with a higher degree of deacetylation (% DD) and with a larger thickness. A transdermal chitosan patch was developed using a chitosan membrane for rate control and a chitosan hydrogel as a drug reservoir. The most prolonged release in vitro was obtained with a 95% DD chitosan rate controlling membrane. The transport mechanism was found to be non-Fickian. The functionality of this transdermal patch was studied on the forearm of human volunteers by assessing the anesthetic effect. Patches with 70% and 95% DD membranes delayed the anesthetic effect, increasing the delay with increasing % DD. It was concluded that a combination of chitosan membrane and chitosan hydrogel is a good transparent system for controlled drug delivery and that the release kinetics in vitro at least for lidocaine have a predictive value for its anesthetic effect in vivo. The demonstration of a direct relationship between in vitro drug membrane permeability and its physiological effect might be considered as quite unique. PMID- 15132183 TI - Gastroretentive delivery systems: hollow beads. AB - The objective of this study was to develop a floatable multiparticulate system with potential for intragastric sustained drug delivery. Cross-linked beads were made by using calcium and low methoxylated pectin (LMP), which is an anionic polysaccharide, and calcium, LMP, and sodium alginate. Beads were dried separately in an air convection type oven at 40 degrees C for 6 hours and in a freeze dryer to evaluate the changes in bead characteristics due to process variability. Riboflavin (B-2), tetracycline (TCN), and Methotrexate (MTX) were used as model drugs for encapsulation. Ionic and nonionic excipients were added to study their effects on the release profiles of the beads. The presence of noncross linking agents in low amounts (less than 2%) did not significantly interfere with release kinetics. For an amphoteric drug like TCN, which has pH dependent solubility, three different pHs (1.5, 5.0, and 8.0) of cross-linking media were used to evaluate the effects of pH on the drug entrapment capacity of the beads. As anticipated, highest entrapment was possible when cross-linking media pH coincided with least drug solubility. Evaluation of the drying process demonstrated that the freeze-dried beads remained buoyant over 12 hours in United States Pharmacopeia (USP) hydrochloride buffer at pH 1.5, whereas the air-dried beads remained submerged throughout the release study. Confocal laser microscopy revealed the presence of air-filled hollow spaces inside the freeze dried beads, which was responsible for the flotation property of the beads. However, the release kinetics from freeze dried beads was independent of hydrodynamic conditions. Calcium-pectinate-alginate beads released their contents at much faster rates than did calcium-pectinate beads (100% in 10 hours vs. 50% in 10 hours). It appears that the nature of cross-linking, drying method, drug solubility, and production approach are all important and provide the opportunity and potential for development of a gastroretentive drug delivery system. PMID- 15132184 TI - The 3D model: explaining densification and deformation mechanisms by using 3D parameter plots. AB - The aim of the study was to analyze very differently deforming materials using 3D parameter plots and consequently to gain deeper insights into the densification and deformation process described with the 3D model in order to define an ideal tableting excipient. The excipients used were dicalcium phosphate dihydrate (DCPD), sodium chloride (NaCl), microcrystalline cellulose (MCC), xylitol, mannitol, alpha-lactose monohydrate, maltose, hydroxypropyl methylcellulose (HPMC), sodium carboxymethylcellulose (NaCMC), cellulose acetate (CAC), maize starch, potato starch, pregelatinized starch, and maltodextrine. All of the materials were tableted to graded maximum relative densities (rhorel, max) using an eccentric tableting machine. The data which resulted, namely force, displacement, and time, were analyzed by the application of 3D modeling. Different particle size fractions of DCPD, CAC, and MCC were analyzed in addition. Brittle deforming materials such as DCPD exhibited a completely different 3D parameter plot, with low time plasticity, d, and low pressure plasticity, e, and a strong decrease in omega values when densification increased, in contrast to the plastically deforming MCC, which had much higher d, e, and omega values. e and omega values changed only slightly when densification increased for MCC. NaCl showed less of a decrease in omega values than DCPD did, and the d and e values were between those of MCC and DCPD. The sugar alcohols, xylitol and mannitol, behaved in a similar fashion to sodium chloride. This is also valid for the crystalline sugars, alpha-lactose monohydrate, and maltose. However, the sugars are more brittle than the sugar alcohols. The cellulose derivatives, HPMC, NaCMC, and CAC, are as plastic as MCC, however, their elasticity depends on substitution indicated by lower (more elastic) or higher (less elastic) omega values. The native starches, maize starch and potato starch, are very elastic, and pregelatinized starch and maltodextrine are less elastic and exhibited higher omega values. Deformation behavior as shown in 3D parameter plots depends on particle size for polymers such as CAC and MCC; however, it does not depend on particle size for brittle materials such as DCPD. An ideally deforming tableting excipient should exhibit high e, d, and omega values with a constant ratio of e and omega at increasing densification. PMID- 15132185 TI - FRONTLINE--the Academy, the APA, and the women. PMID- 15132186 TI - Changing patterns of femininity: psychoanalytic implications. AB - The social status and gender-role behavior of women has changed periodically over history, depending on a variety of socio-cultural, economic, and religious factors. Classical psychoanalytic views concerning "normal" feminine psychology and sexuality, based on Freud's original postulates were derived from his experiences and observations as a late-19th and early-20th century middle European male. This article explores the errors involved in these traditional hypotheses concerning "penis envy," "normal" feminine masochism and passivity, female super-ego development and female psychosexuality, including orgasm and gender-role behavior. PMID- 15132187 TI - Mysteries of the void: an analyst's view of female urination. AB - Anxiety, shame, guilt, and depression accompany the absence or loss of urinary sphincter control at both ends of the life cycle. The economic costs are high when one reflects that incontinence limits access to child-care facilities and thus the mothers return to gainful employment. It is often the prime reason for admission to nursing home care for the grandmother. Poise and increased self esteem go along with "being dry." Dysuria accompanying acute or interstitial cystitis also interferes with personal peace and the ability to work productively. A psychoanalytic review of developmental stages and their constellation of affects and defenses in regard to "wetness" will be discussed as well as when to refer, clarify, or make or receive a urological referral for your patient, their child, or parent. PMID- 15132188 TI - On being an old woman: a personal perspective. AB - This is a personal, point-of-view article that speaks to the daunting task of living well when one is old, with a particular focus on a woman in our society. Issues of both loss and challenge are addressed. The article concludes that how a woman leads her entire life will either prepare her for old age or it will fail to do so. Society will not do for her what she has failed to do for herself. PMID- 15132189 TI - The hand that rocks the cradle rocks the boat: the empowerment of women. AB - The obstacles to equality for women have changed from external to intrapsychic. The relational characteristics of women, such as nurturing and connecting with others, are now regarded as desirable attributes for the workplace. Adult growth and achievement models are based on the so-called masculine characteristics of separation, individuation, and independence. Relational theory supplies an alternative model called growth-in-connection, which stems from the feminine characteristics of connection, collectivity, and interdependence. Women's fears of exercising power, men's resistance to sharing power, the traditional ways in which women attain power are all concerns that are examined. Real power lies in exercising leadership, which implies risk-taking. Exploring the gender disequilibrium in issues of power--both the female advantage and the female disadvantage-as exemplified by the glass ceiling and its origins--helps to clarify the issues involved in empowerment. PMID- 15132190 TI - Personal power and the cultural unconscious: implications for psychoanalytic theories of sex and gender. AB - The changes in the way psychoanalysts theorize female psychology was in large part the result of changes that occurred in the context of the sex and gender movements of the last half of the 20th century. This article addresses the covert ways in which women sought to express power prior to women's liberation. It draws on psychoanalytic theory, emphasizing the study of power in conjunction with discourse theory in order to illustrate the complex ways change draws on both inner psychic impulses and changes in the cultural milieu. PMID- 15132191 TI - The psychodynamics of patients with false memory syndrome (FMS). AB - The False Memory Syndrome (FMS) diagnosis was very much in vogue from the late 1980s to the mid-1990s. It was an outgrowth of the belief held by many therapists that childhood sexual abuse was one of the most common causes of all forms of psychopathology. Although no longer in-vogue, the diagnosis is more recently being used for people who are falsely claiming that they were sexually abused by their priests. As was true in the earlier era, there were indeed many people who were sexually abused in childhood, but there were also many who were not and actually came to believe that they were, especially under the influence of therapists conducting "repressed memory therapy." Similarly, sexual abuse by priests has been a widespread phenomenon. Yet, there are still false accusers who are being led to believe by their overzealous therapists that they were indeed abused. As a psychoanalyst who has been asked to do forensic evaluations in many of these cases, I automatically ask myself questions about the psychodynamics of such falsely accusing patients. Here I describe those psychodynamic factors that I believe were operative in FMS cases, factors which in some cases apply to false sex abuse accusation against priests. Accordingly, this article is not simply of historical interest, but is still relevant and timely. PMID- 15132192 TI - Adult mother-daughter relationships: a review of the theoretical and research literature. AB - This review of the psychoanalytic, developmental, and other relevant theoretical and research literature on mother daughter relationships was undertaken as part of an ongoing research study, Generation to Generation. Mother-Daughter Physicians (Shrier and Shrier 2000, 2002b). The review focuses particularly on mothers and their adult daughters during the longest period of a woman's life (between the end of adolescence and old age). The research literature on normative mother-daughter relationships is quite limited and while there is an extensive and evolving theoretical literature, it is not grounded in a nonclinical empirical database. Highlights are presented of past and current theories and research studies about female development and mother-daughter relationships, embedded in their historical sociocultural context. This comprehensive literature review documents the need for an empirical database drawn from a nonclinical population and the importance of methodologically sound research to support or challenge existing and evolving developmental and psychoanalytic theories on adult mother-daughter relationships. PMID- 15132193 TI - Having it all: the professional mother's dilemma. AB - The notion that professional women can "have it all"--career and children--has been disputed in the current popular literature. A brief review of recent findings presents evidence that many couples successfully manage the dual challenge of work and family. Clinical vignettes from a group of professionals/mothers are presented with suggested "rules" for success. We have come a long way from the days when it was taken for granted (mostly by our grandfathers) that women's place is only in the home. PMID- 15132194 TI - Breaking through the despair: spiritually oriented group therapy as a means of healing women with severe mental illness. AB - Studies have shown that 96% of Americans believe in God and over 90% pray yet there is relatively little education available for clinicians on how to use spirituality as a tool for healing mental illness, particularly when treating very sick patients. This article illustrates how spiritually oriented group therapy with severely ill women can help to improve mood, affect, motivation, interpersonal bonding, and sense of self, and can succeed in reaching patients and promoting recovery in ways that traditional therapy cannot. Specific modalities including group prayer, yoga breathing, and spiritual readings are described. Breaking Through the Despair offers both a psychodynamic and a neurophysiologic perspective for understanding how this type of treatment helps patients transcend their mental illness and be able to grasp abstract spiritual concepts, develop a sense of belonging to a caring community, and integrate a new sense of themselves as productive and valued individuals. PMID- 15132195 TI - Medieval orality, mothers, and bonding. AB - The role of women in the Middle Ages was vilification, veneration, and exclusion. Due to the high rates of maternal and infant mortality bonding shifted from the mother-child dyad to one in which the Church, Holy Family, and king acted as pseudo-parents. In art this is suggested by the virtual absence of eye contact between the Virgin and Christ-child. Frustration of early oral needs consequent to lack of adequate mother-child bonding prompted a reactive emphasis on orality in art and legend. A decrease in infant mortality and a reciprocal improvement in mother child bonding contributed to cultural shifts in how self-realization would be accomplished during the Renaissance and in the later emergence of secular humanism. PMID- 15132196 TI - Artemisia Gentileschi and her world. AB - This article deals with one of the most notable artists of 17th century Italy: Artemisia Gentileschi. Gentileschi holds a unique place in art history, both as a woman artist and as the first female member of the Academy of Design in Florence. Brought up to be an artist by her father--the painter Orazio Gentileschi--she was allowed to work in his studio, use models, receive instruction and collaborate with well-known artists. Artemisia Gentileschi became a follower of the school of Caravaggio--the school's only Caravaggista. She began public life in a notorious way; her father had his colleague Tassi--her painting instructor--charged with raping her. The relationship between historical and art historical events in Gentileschi's lifetime and creative processes are explored. Special attention is given to those vicissitudes of Gentileschi's personal life that may be reflected through her work. Popular psychoanalytic conceptualizations about Gentileschi are discussed, as are mythological and biblical themes in her art. PMID- 15132197 TI - Women's bodies, doctors' dynamics. AB - Medicine treats men's and women's reproductive organs and functions markedly differently. Interventions affecting male functioning are regarded as very serious and are seldom undertaken except to treat significant pathology. Interventions affecting perfectly normal female functioning, such as treatment for menstrual cycle mood changes, menopause, episiotomy, cesarean and other assisted delivery, hormonal contraception, and even hysterectomy, are rampant. These interventions can be attributed to men's unconscious (and conscious) curiosity, jealousy, hostility, and helplessness about female reproduction, coupled with the fact that men have dominated the field of medicine for most of its history. This article describes these interventions and the rationalizations for them, and questions their utility and meaning in the light of current knowledge. PMID- 15132198 TI - Gender in the consulting room. AB - Gender influences psychoanalysis and psychotherapy in a variety of ways. This article discusses these with reference to the woman therapist and analyst. Choice of therapist is influenced by realistic, transferential, and stereotyped ideas such as (1), wishes for a role model, (2) unconscious fantasies for a better mother, and (3) ideas that women are more nuturent. Family and work conflicts, pregnancy, and lifestyle issues are special issues for the woman professional. Transference and coutertransference manifestations concern variations in erotic and eroticized transference, maternal and paternal transference, and paternal erotic countertransference. Some limitations of cross gender treatment are indicated. PMID- 15132199 TI - The feminization of psychiatry? Some ruminations. AB - This article considers the position of women in psychiatry today from the historical perspective of feminism. Feminism in medicine demands the inclusion of the traditional priorities of women: collaboration and cooperation over competition and hierarchy, compassionate care over technology and automation, flexible care for the individual in a social context over the study of units of diseased organs. These themes, in addition to the care of women and children, were prominent in the lives of early women physicians and again in the 1970s and 1980s. The number of women in psychiatry has increased. However, their influence is scant and feminist goals are again submerged. At the same time psychiatry has become increasingly interested in the organ (brain) at the expense of the individual person. Women need a new awakening. They must use their new presence to assert feminine values in patient care and protect themselves from becoming provider units on the assembly line. PMID- 15132200 TI - Balinese women in a changing society. AB - Balinese women face the dilemma of maintaining their vital role amid a rapidly changing society. In Bali, the primary female role is one of fostering balance and harmony within families. The Balinese people view women not from the vantage of career success but rather from the vantage of whether they can produce good quality children, and can work as part of a family team. Balinese men and women work together as partners. Indeed, men are not enemies; the genders help and need each other. Values underlying emancipation for women clash with traditional values, leading to frequent misunderstandings. Emancipation advocates neglect those elements necessary for complementing Balinese values. Applying educational and preventative methods, as well as therapeutic innovations to such problems, is helpful at all levels of society. PMID- 15132201 TI - Studies in countertransference and gender: female analyst/male patient in two cases of childhood trauma. AB - The paternal transference and the erotic transference have been singled out in the literature as more problematic for women analysts with male patients. Two clinical cases provide evidence for their appearance indicating that the reactions they trigger in women may explain the difficulties in their recognition and interpretation. In these particular cases, the author analyzes her counterresistance to a projected narcissistic, cruel father object and her defense of masochistic submission, following more stereotypical gender lines. And in the case of an eroticized transference, the fears of a symbiotic and incestuous merger made more difficult the elucidation of the complex transference reaction. The hypothesis is forwarded that cultural prohibitions are partly responsible for the lack of recognition of these transferences and for the challenging countertransference that they elicit. PMID- 15132202 TI - Restyling our lives to reduce cardiovascular disease risk: what will it take? PMID- 15132203 TI - Religion, spirituality, and healthcare choices of African-American women: results of a national survey. AB - OBJECTIVE: This study describes the prevalence and patterns of use of religion and spirituality for health reasons among African-American women. METHODS: Respondents were asked about their use of religion/spirituality for health reasons as part of a larger study of the prevalence and correlates of complementary and alternative medicine (CAM) use among women. In 2001, a national survey of 3,172 women, aged 18 and older, was conducted in 4 languages, with over sampling among African-, Mexican-, and Chinese-American participants. This paper focuses on the sub-sample of 812 African-American women. RESULTS: Overall, 43% of the African-American women reported using religion/spirituality for health reasons in the past year. Factors significantly associated with the use of religion/spirituality for health reasons included having an income of dollar 40,000-dollar 60,000, an education level of college graduate or more, or being 37 56 years of age; worse health status approached significance. African-American women utilized religion and spirituality most often for serious conditions such as cancer, heart disease, and depression. African-American women who had used religion/spirituality in the past year for health reasons were more than twice as likely to have used some form of CAM, and also more likely to have seen a medical doctor during the year prior to the interview, compared to their counterparts. CONCLUSION: Religion and spirituality are associated with health-seeking behaviors of African-American women. The use of religion and spirituality for health reasons warrants additional research, particularly its use for chronic and serious conditions, and its role in the health-seeking behavior of African American women in conjunction with the utilization of conventional medicine and CAM. PMID- 15132204 TI - Correlates of physical activity levels in a sample of urban African Americans with type 2 diabetes. AB - BACKGROUND: Although regular physical activity is recommended for all adults and is vital in the management of diabetes, activity levels among African Americans with diabetes continue to be sub-optimal. The factors influencing physical activity in this group have not been well examined. RESEARCH DESIGN AND METHODS: Physical activity levels were assessed in 186 African Americans with type 2 diabetes in an urban inner-city community in 4 daily domains; leisure-time physical activity, episodic vigorous activity, blocks walked, and stairs climbed. Linear and logistic regression techniques were used to identify factors independently associated with physical activity levels. RESULTS: A minority of both men (40%) and women (29%) reported engaging in regular physical activity for the purpose of exercise. Women walked significantly fewer blocks/week compared to men (17 vs 41, P < .05). Independent predictors of low physical activity were obesity, lower household income, and the self-perception of being more active than one's counterparts. A predictor of higher physical activity was the perception of needing to get enough exercise to keep healthy. There was no association between physical activity level and other characteristics, including the perception of oneself as overweight or trying to lose weight. CONCLUSIONS: These data suggest that, among our sample of African Americans with diabetes, many do not engage in regular activity, women walk significantly less than men, and weight loss efforts may not commonly include physical activity. Obesity, lower income level, and confidence about activity levels may identify individuals with lower physical activity levels. These data should be useful for developing targeted and culturally appropriate interventions to promote physical activity in this high-risk community. PMID- 15132205 TI - Genetic and environmental influences on forced expiratory volume in African Americans: the Carolina African-American Twin Study of Aging. AB - OBJECTIVES: Previous research found measures of pulmonary functioning to be strong predictors of cognitive functioning and mortality; however, there is considerable individual variability in performance on these measures. In the present analyses, the relative contribution of genetic and environmental influences to variability in average peak expiratory flow rate (APEFR) are examined in a sample of adult African-American twins. DESIGN: Birth records from North Carolina Register of Deeds offices were used to identify participants for the Carolina African-American Twin Study of Aging (CAATSA). Participants completed an in-person interview, which included measures of health status, cognition, and psychosocial measures. PARTICIPANTS: Data for the analysis come from 200 pairs of same sex twins (97 identical pairs, and 113 fraternal), with a mean age = 46.9 years (SD = 13.9), and with 39% of the sample being men. RESULTS: Phenotypic correlations between APEFR, age, gender, height, and cigarette consumption (measured in pack years), were all significant, ranging from -.63 to .43. After the affects of age, gender, height, and pack years were partialled out of APEFR, quantitative genetic analyses were conducted on the residuals. Model fitting demonstrated that variance in APEFR was accounted for by shared environmental effects (30%), genetic effects (14%), and non-shared environmental effects (56%). CONCLUSION: These results are discussed in relation to previous research conducted in other countries, and the importance of a complex systems approach to explanations of the impact of genes on central indices of health, such as APEFR. PMID- 15132206 TI - A review: maximizing social support--a neglected strategy for improving weight management with African-American women. AB - Although research has identified various factors that can affect African-American women's weight management efforts, limited attention has been paid to the role of social support. Culling evidence from heretofore disparate studies, this article provides an overview of why adequacy of social support may be among the most critical factors affecting African-American women's weight management behaviors. The empirical evidence suggests that social support can enhance perceived control and/or self-efficacy, and this finding serves as a focal point for looking at aspects of weight loss programs, as well as providing a post-program strategy, in order to maximize this support among African-American women. PMID- 15132207 TI - Dietary differences among women of Polish descent by country of birth and duration of residency in the United States. AB - OBJECTIVES: Changes in breast and gastric cancers and coronary artery disease among people of Polish descent after migration to the United States suggest there may be potentially modifiable factors affecting incidence of these diseases. We examined relationships of dietary factors associated with these diseases with stage of migration among Polish women in Chicago. DESIGN: Women of Polish descent (N = 396) were selected from Polish women's social organizations. Women completed a modified Health Habits and History Questionnaire. SETTING: The questionnaire was completed either at the participant's home or at a Polish social organization. PARTICIPANTS: Participants ranged in age from 17-81 years, and included women born in Poland or the United States, who had at least one parent of Polish. INTERVENTIONS: Participants were stratified by country of birth and migration period (1935-1979, 1980-1989, 1990-1997). MAIN OUTCOME MEASURES: The average daily intake of food groups and nutrients was assessed using multiple linear regression. RESULTS: We found statistically significant differences by birth country for 19 of 34 nutrients, 4 of 7 food groups, and for 21 nutrients, and 5 food groups among the different migration tertiles. CONCLUSIONS: Women from Poland and more recent migrants had generally more nutritious intakes, compared to US-born women, or earlier migrants. APPLICATIONS/CONCLUSIONS: There are significant dietary differences among women of Polish descent that vary by duration of US residency and birth country. Women with dietary intakes which place them at higher risk for cancers and cardiovascular disease could be targeted for interventions to lower their disease risk. PMID- 15132208 TI - Correlates of femoral artery flow mediated dilation in a multi-ethnic sample of 12- to 26-year-olds. AB - OBJECTIVE: Endothelial dysfunction is one of the earliest events in the pathogenesis of cardiovascular disease (CVD). Studies involving healthy adults have found endothelial function, measured via flow mediated dilation (FMD), to be impaired in African Americans (AAs) compared to European Americans (EAs). The purpose of this study was to determine whether ethnic differences exist in FMD in a group of healthy teenagers and young adults, and to examine separately, by ethnic group, the relationships between FMD and several measures of adverse cardiovascular prognoses. DESIGN: Subjects underwent measurement of various anthropometric, hemodynamic, and echocardiographic variables. SETTING: Measurements were made in a laboratory setting. PARTICIPANTS: Subjects were 159 12- to 26-year-old AAs and EAs. MAIN OUTCOME MEASURES: FMD, endothelin-1 (ET-1) levels, at rest and in response to stress, and measures of cardiac structure and function. RESULTS: Ethnic differences were not observed in FMD. Only pre occlusion arterial diameter was significantly related to FMD in EAs (P < .001); whereas, among AAs, FMD was negatively correlated (all Ps < .03) with gender, pre occlusion arterial diameter, relative wall thickness (RWT), resting plasma ET-1, and ET-1 increases, related to a behavioral stressor. Multiple linear regressions for AAs revealed that gender and RWT each explained a unique variation in FMD (Total model R2 = .36, P < .0001). CONCLUSION: Ethnic differences in FMD are not in evidence for subjects in this age range. The inverse relationships observed in AAs between FMD and measures of altered endothelial system function, and ventricular structure and function, provide additional evidence for early clustering of measures of adverse cardiovascular prognoses in this group. PMID- 15132209 TI - The global cardiovascular diseases risk pattern in a peri-urban working-class community in South Africa. The Mamre study. AB - OBJECTIVES: To describe the cardiovascular disease (CVD) risk factors and the global burden of CVD risk in a peri-urban, working-class community of Mamre near Cape Town. To identify additional variables in the data set associated with the global CVD risk factor score. The latter was calculated using the major CVD risk factors in formulas derived from the Framingham global CVD risk calculations. Such variables could possibly be used for global CVD risk calculations, instead of depending on biochemical estimates for these calculations. METHODS: In a random population-based sample of 976 people aged 15 years and older, data on demography, smoking, physical activity, and alcohol use were collected. Blood pressure (BP), anthropometry, levels of serum glucose and lipids, and low-density lipoprotein cholesterol (LDL) particle sizes were also determined. These data allowed calculation of the global CVD risk profile with the Framingham study's formula. The data are age-standardized to the colored (mixed ancestry) population according to the 1996 South African census. RESULTS: The global CVD risk score suggested that men and women had a 5.2% and 4.2% probability, respectively, of having a CVD event in the next 10 years, while for those 55 years of age and older, the probability increased to more than 30% and 25%, respectively. Hypertension was found in 22% of men and 16% of women. Sixty-two percent of the men and 44% of the women smoked cigarettes, while 6% and 5% had diabetes, respectively. Hypercholesterolemia was present in 47% of men, and 46% of women. Small-dense LDL particles were present in 26% of men and 14% of women. A number of easily measured CVD risk factors could explain 40.3% of the variation of the global CVD risk score. These include aspects of the medical history provided by the patient, the inverse of the amount of physical activity and weight measurements, as well as height, and waist circumference. CONCLUSIONS: The people in Mamre have a high probability of suffering a CVD event in the next 10 years. Age and gender are the primary contributors to the global CVD risk score. The findings suggest the possibility of developing a global CVD risk score based on easily measured CVD risk factors for use in developing countries with limited resources. PMID- 15132210 TI - Characteristics of ketosis-prone diabetes in a multiethnic indigent community. AB - OBJECTIVE: To compare demographic and clinical characteristics among 3 ethnic groups of indigent patients exhibiting diabetic ketoacidosis (DKA), in Houston, Texas. METHODS: Over a span of 3.5 years, 321 patients were interviewed at the time of admission for DKA. Demographic, clinical, and biochemical data and measures of pancreatic beta-cell function were obtained at baseline and during follow up. Pearson's chi-square test, or one-way ANOVA, were used, as appropriate, to evaluate group differences. RESULTS: Of the 321 subjects, 44% were African-American, 40% were Hispanic, and 16% were Caucasian. A significantly higher proportion of Hispanics had preserved beta-cell function, compared to African Americans and Caucasians (51% vs 32% and 32%, respectively; P = .002). This difference, present at the time of the admission, was maintained through follow up. In a multivariate analysis, Hispanic ethnicity (OR 3.61; 95% CI 1.48 9.29) was a significant predictor of preserved beta-cell function. In addition, Hispanics were less likely to develop DKA as a result of treatment non compliance, and more likely to have DKA precipitated by an acute illness. CONCLUSIONS: Our findings indicated that ethnicity is associated with significant differences in the pathophysiologic and clinical characteristics of indigent, ketosis-prone patients. Hispanic ethnicity was found to be associated with greater beta-cell functional reserve, and less dependence on chronic insulin therapy. PMID- 15132211 TI - Prevalence of hypertension in some cross-cultural populations of Visakhapatnam district, South India. AB - Hypertension is becoming a major public health problem in developing countries. Blood pressure tends to vary cross-culturally and, as yet, none of the studies from India, particularly those from the state of Andhra Pradesh, has attempted to address the influence of acculturation/modernization on the prevalence of hypertension. The present study investigates the prevalence of hypertension in some cross-cultural populations of Visakhapatnam district, Andhra Pradesh, India, and to report the influence of acculturation/modernization on hypertension prevalence. One thousand three hundred sixteen individuals (646 men and 670 women), belonging to 2 tribal and 2 caste groups (from both rural and urban areas), were surveyed for hypertension prevalence. The recommendations of JNC-VI were utilized to classify participants by different categories of hypertension. A substantial proportion (21%) of the individuals exhibited various stages of hypertension. There are considerable differences between these cross-cultural populations, particularly between tribes and castes. The results confirmed the hypothesis that acculturation/modernization may elevate the risk of hypertension, and that prevalence is generally low among traditional population groups. The risk of hypertension increased for those aged 45 years and older, and this risk was greater for women, compared to men. PMID- 15132212 TI - International collaborative study of cardiovascular disease in Asia: design, rationale, and preliminary results. AB - OBJECTIVES: The overall objective of the International Collaborative Study of Cardiovascular Disease in Asia (InterASIA) was to estimate the prevalence and distribution of cardiovascular disease risk factors, including hypertension, hypercholesterolemia, diabetes, obesity, cigarette smoking, and physical inactivity, in the general population of both China and Thailand. METHODS AND RESULTS: A multistage cluster sampling method was used to select a nationally representative sample of 15,838 adults, aged 35-74 years, in China, and 5,350 similarly aged adults in Thailand. Data on medical history, cigarette smoking, alcohol consumption, diet, physical activity, and awareness and control of risk factors for cardiovascular disease, were collected. Blood pressure, body weight and height, and hip and waist circumferences, were measured using standard methods. Fasting blood specimens were collected to measure levels of total cholesterol, high-density lipoprotein cholesterol, triglycerides, glucose, and creatinine. All data were carefully collected by specially trained, experienced observers using standardized methods and a stringent level of quality control. The prevalence of self-reported cardiovascular disease in Chinese adults aged 35 74 years was 3.3% in men and 3.6% in women (representing 8,147,000 male and 8,270,000 female cases nationally). The prevalence of self-reported cardiovascular disease in Thai adults was 1.5% in men and 1.7% in women (representing 184,000 male and 226,000 female cases). CONCLUSIONS: The InterASIA study provides important information on the current prevalence and control of both cardiovascular disease and its risk factors in the region. These findings will be useful for developing national strategies to reduce the large, and increasing, burden of cardiovascular disease in China and Thailand. PMID- 15132213 TI - The feasibility of partnering with African-American barbershops to provide prostate cancer education. AB - The purpose of this study was to determine the feasibility of partnering with barbershops to implement a community-based prostate cancer preparatory educational decision aid for African-American men. We used African-American newspapers to compile a list of barbershops in King County, Washington, that predominantly serve African-American men. Trained research interviewers conducted proprietor surveys and client surveys. Ninety-six percent of the proprietors surveyed reported they would allow their clients to learn about prostate cancer. Seventy-five percent reported they would consider allowing a computer to be installed to provide information about prostate cancer. Ninety-seven percent of clients reported that they would be willing to look at information about prostate cancer in their barbershops. It will be feasible to work with barbershops and their clients for a community-based prostate cancer screening decision-aid intervention for African-American men. PMID- 15132214 TI - Smoking cessation and its determinants among older American Indians: the Strong Heart Study. AB - OBJECTIVE: To examine the relationship between sociodemographic, clinical, and smoking history factors, and smoking cessation among older American Indians. DESIGN: Nested cohort study of cigarette smokers in the Strong Heart Study, a longitudinal study of cardiovascular disease among American Indians. SETTING: Thirteen American Indian tribes from Arizona, Oklahoma, and North and South Dakota. PARTICIPANTS: American Indian men and women (N = 998), aged 45-74 years, who identified themselves as smokers at the initial Strong Heart Study examination. MEASUREMENTS AND MAIN RESULTS: Twenty-one percent of smokers quit during the 4-year follow-up period. Multivariate logistic regression was used to assess the relationship between baseline sociodemographic, clinical, and smoking history factors, and smoking cessation. Factors associated with smoking cessation included being 65-74 years old (odds ratio [OR] 2.1; 95% confidence interval [CI], 1.3 to 3.3), being examined at the Arizona regional center (OR 2.2; 95% CI 1.3, 3.7), being non-daily smokers (OR 5.4; 95% CI 1.3, 18.5), smoking fewer than 6 cigarettes daily (OR 2.8; 95% CI 1.3, 4.7), being a smoker for fewer years(OR 2.0; 95% CI 1.0, 3.9), beginning to smoke at an older age (17 years or older, OR 1.6; 95% CI 1.1, 2.4), and having a history of diabetes (OR 1.7; 95% CI 1.2, 2.3). Factors not associated with smoking cessation included gender, level of education, childhood exposure to tobacco smoking, and a history of cardiovascular diseases, cancer, or respiratory diseases. CONCLUSION: Several determinants of smoking cessation among older American Indians identified in this study may have important implications for designing appropriate interventions for this special population. PMID- 15132215 TI - Variations in hip fracture hospitalization rates among different race/ethnicity groups in New York City. AB - Hip fractures are a major cause of death and disability among the elderly. Although previous studies have shown hip fracture rates to be highest among Whites, and lowest among Blacks, less is known about how these differences are affected by gender and age. We determined the hip fracture hospitalization rates for non-Hispanic Whites, Blacks, Hispanics, and Asians in New York City from 1988 to 2002 using Statewide Planning and Resource Cooperative System data from the New York state Department of Health. We found that hip fracture hospitalization rates for those aged 50 and older were higher for women than men, with a risk ratio of 1.75 (95% confidence interval [CI] 1.73-1.77). Annual age-adjusted hip fracture hospitalization rates among White, Black, Hispanic, and Asian women were 459, 137, 143, and 174 per 100,000, respectively. The corresponding rates for men were 230, 109, 87, and 104, respectively. For both genders, and all race/ethnicity groups, the rates increased sharply with age. Although overall, women had higher rates compared to men, these differences varied by race and age. For Whites and Asians, women had higher rates for all age groups older than 50 years. For Hispanics aged 50-59, men had a higher rate than did women, but this gender relationship reversed after age 60. For Blacks, however, men had higher rates than women until age 70, after which women had higher rates. These results demonstrate that race/ethnicity, as well as age and gender, influence the incidence of hip fractures. Moreover, Asians, with their low bone mass, showed no increased hip fractures, compared to non-Hispanic Blacks and Hispanics. Further study is needed to explain this paradox. PMID- 15132216 TI - Compliance with recommendations for follow-up care in Latinos: the Los Angeles Latino Eye Study. AB - PURPOSE: To determine rates of follow-up eye and health care in the Los Angeles Latino Eye Study (LALES), a population-based sample of Latinos. METHODS: Participants received a complete ophthalmic examination and were referred to a local healthcare provider for follow-up care, if ocular or systemic disease was found. Participants receiving referrals were later contacted by telephone and interviewed in their language of choice (English or Spanish) by a trained bilingual interviewer, to determine follow-up rates, and to discuss the barriers preventing follow-up care. RESULTS: Of 430 referred participants, 335 (78%) completed the follow-up survey; 278 (68%) of the responders obtained follow-up care. Among the 108 (32%) individuals who did not seek follow-up care, 54 (50%) cited cost of care as the main reason, while 30 (28%) indicated a lack of knowledge as to where to go for care, and 18 (17%) indicated the unavailability of health care. Logistic regression analyses (controlling for acculturation, co morbid conditions, and patients' prior knowledge of their diagnoses) revealed that participants with insurance coverage, diagnosis with a systemic disease, and higher educational level were more likely to have received follow-up care. CONCLUSION: More than two thirds of the participants reported seeking the recommended follow-up care. Although cost, availability of services, and convenience of accessing care were found to be major barriers to obtaining health care, higher education, insurance coverage, and prior knowledge of the disease, were associated with receiving recommended care. Knowledge of barriers to seeking health care is important when developing community based healthcare programs directed toward the Latino population. PMID- 15132217 TI - Quality of birthplace information obtained from death certificates for Hispanics, Asians, and Pacific Islanders. PMID- 15132218 TI - For the patient. Identifying risk-factor patterns for heart disease in South Africa. PMID- 15132219 TI - For the patient. Barbers as partners to deliver prostate cancer education. PMID- 15132220 TI - For the patient. African-American women: your social groups can improve weight management. PMID- 15132221 TI - For the patient. Diets change as new customs are adopted. PMID- 15132222 TI - For the patient. Characteristics of ketosis-prone diabetes in a multiethnic indigent community. PMID- 15132223 TI - Military medicine, its technology and transition in the mid-twentieth century. PMID- 15132224 TI - Prostate cancer screening among military service members. AB - This retrospective study investigated the characteristic of military male beneficiaries age 50 years and older who were screened for prostate cancer at a medical center in the Pacific Regional Command. Data were collected on male patients who were seen in three adult outpatient clinics from December 1999 through November 2001. Two research questions were asked: What are the characteristics of men age 50 and older who were screened for prostate cancer? Are there variations in prostate cancer screening based on age, ethnicity, and military status? Data were analyzed using descriptive statistics. The sample was predominantly Caucasian and Asian/Pacific Islander who were over age 67. Although there was evidence of screening in 68% of the sample, 13% of the men had been screened according to the American Cancer Society's recommendations. The findings support the need for a prostate cancer screening policy for the Department of Defense. PMID- 15132225 TI - West Nile virus. AB - West Nile virus infection has quickly become a feared cause of neurologic disability and death, particularly when it presents with encephalitis. Recent epidemics in endemic regions of Eurasia and Africa, as well as its recent spread to North America, have highlighted the need for all physicians to be aware of its clinical presentation and course. In particular, because of the increased susceptibility of West Nile virus infection during outdoor activities, as well as during travel to the Middle East and Southeastern Europe, military physicians should be informed about case recognition, management, and prevention to maintain the health of soldiers and their families. PMID- 15132226 TI - Wound shock: a history of its study and treatment by military surgeons. AB - The treatment of wounds has received considerable attention from the time of the Trojan War. However, it was not until the American Civil War that shock was described as an entity distinct from the wounds themselves and that efforts were directed at more than just treatment of the wound. The need for fluid resuscitation in the treatment of hemorrhagic shock was first recognized in the Spanish American War, as was the association of sepsis with shock. World War I showed the need for blood in the treatment of "wound shock," a lesson that had to be relearned in World War II through bitter experience. Studies in the Korean War described the concept of disseminated intravascular coagulation and multiple organ failure, and the existence of disseminated intravascular coagulation was confirmed by studies in Vietnam. The treatment of hemorrhagic shock is now very effective, but the treatment of traumatic and septic shock remains unsatisfactory. PMID- 15132227 TI - Evaluation of oral or rectal midazolam as conscious sedation for pediatric patients in oral surgery. AB - This study was undertaken with a view to determine the acceptance and treatment possibilities of midazolam, depending upon its oral and rectal application for pediatric patients requiring an oral surgery procedure (tooth extraction) and having a pretreatment behavioral score of 1 or 2 according to the Frankl Scale. Oral (0.5 mg/kg) and rectal (0.35 mg/kg) midazolam was compared in view of acceptance of the mode of treatment and local anesthesia, level of amnesia, and adverse effects. Although oral or rectal midazolam application has similar characteristics in respect to ease of working, the oral midazolam application should generally be preferred because it is more easily accepted by pediatric patients. PMID- 15132228 TI - Thalassemia screening among Royal Thai Army medical cadets. AB - Thalassemia, common in Thailand, varies from mild to severe anemia, resulting in work inefficiency, particularly during exertion. Therefore, it is important for military cadets to be screened. The objective of this study was to screen for thalassemia and hemoglobinopathies among Royal Thai Army medical cadets. We tested 358 third-year Royal Thai Army medical cadets for complete blood count, red blood cell indices, hemoglobin (Hb) typing, inclusion bodies, and Hb A2, and Hb E. DNA analysis confirmed alpha-thalassemia, trait detection. The Hb E trait was the most frequent, respectively, in men and women, at 12.61% and 12%, followed by the a-thalassemia1 trait, at 3.3% and 4% and the beta-trait, at 1.5% and 0%. Two cases of homozygous Hb E were found only in men. Interestingly, a mild form of 1-thalassemia/Hb E was found in one male RTA medical cadet. These findings suggest that consistent thalassemia screening should be considered. PMID- 15132229 TI - Ocular laser exposure incident reporting. AB - BACKGROUND: Advances in laser technology are providing opportunities for new laser applications. Once the nearly exclusive province of scientists and the military, lasers are finding increasingly widespread applications in modern life. Along with this increase in applications, it is reasonable to expect a concomitant increase in the number of accidents and occasions of purposeful misuse, resulting in excessive ocular exposures to laser radiation. METHODS: A survey was conducted of databases currently collecting laser exposure and injury information. Those found were then evaluated for content, organization, and ease of use. RESULTS: A number of laser injury databases, privately and publicly funded, were found Overall, information on laser-associated injuries was quite fragmented, showing significant variability in format and content. CONCLUSIONS: Public health policy development and clinical management of eye injuries would benefit from a single, comprehensive, and easily accessible database/information tool such as the U.S. Army's Laser Accident and Incident Registry. PMID- 15132230 TI - The patient as a client: a model for evaluation of Israel Defense Forces. AB - The Israel Defense Forces (IDF) Medical Corps provides medical services-routine and emergency--to all IDF personnel (conscripts, career personnel, and reservists). Despite the fact that there are no differences in prevalence of disease in the IDF compared with the civil sector, health consumer appraisal, in the military, of the medical services they receive during peacetime at IDF medical clinics is not high. The objective of this research was to develop an effective and differential tool for monitoring the quality of medical service at IDF clinics drawn from service quality indexes based on the perspective of the soldier patient. The research tool used was an anonymous questionnaire comprised of five demographic questions and 21 components of quality index measurements that participants were asked to rank in terms of importance during a visit to their IDF medical clinic. Those categories found to influence the client's perception of quality medical service were accessibility and availability of services, information provided by the clinic, and the efficiency of staff. The factors found to be less influential in the clients' perceptions of quality were staffing and infrastructure. Quality control using these significant indexes will allow monitoring programs to focus on components that are important from the soldier's perspective, without overlooking other significant aspects of the soldier's perceptions of the quality of medical service as a client. PMID- 15132231 TI - Lethal injuries among the members of the 4th Guardian Brigade of Croatian Army during the 1991-1995 war. AB - OBJECTIVE: We analyzed the causes of deaths among the members of the 4th Guardian Brigade (GB) of the Croatian Army during the war in Croatia from 1991 to 1995: the site of the lethal injuries, the type of wounds, and estimated the severity of injuries with lethal outcome according to the Abbreviated Injury Scale. METHODS: This was a retrospective study using the files and data obtained from 4th GB, Croatian Ministry of Defense, and Croatian Ministry of War Veterans. RESULTS: During the War in Croatia from 1991 to 1995, 182 members of 4th GB were killed. One hundred fifteen (63.2%) suffered lethal injuries caused by shell fragments, 47 (25.8%) soldiers had gunshot wounds, and 20 ( 11.0%) died in traffic accidents. Mean Abbreviated Injury Scale for killed soldiers was 7.61 +/- 1.27. CONCLUSION: During the war in Croatia, the leading causes of death were mines and explosions, and, in a minor proportion, gunshot wounds. PMID- 15132232 TI - Evaluation of a contingency blood donor program on U.S. Navy submarines. AB - OBJECTIVE: Because the role of submarine warfare is shifting from strategic deterrence to littoral force projection, submarine providers will be required to manage combat trauma. Currently, submarine providers have only crystalloid available for the reversal of hemorrhagic shock. A proposal program to provide blood products on submarines was evaluated. METHODS: Existing military emergency blood transfusion protocols were reviewed. The restrictions of donation/transfusion onboard submarines were considered. RESULTS: A protocol to provide screening for, implementation of, and a reporting system for contingency blood donation and transfusion onboard submarines was created. The protocol contains all the safeguards of existing U.S. Navy contingency donor programs with the exception of pretransfusion infectious disease testing and cross-matching. However, because the program does not require laboratory capability, it can be implemented by an independent duty corpsman onboard a submarine. CONCLUSION: This protocol provides blood for the reversal of hemorrhagic shock onboard submarines in the event of traumatic injury. PMID- 15132233 TI - Fatal methotrexate-induced pneumonitis in a psoriatic patient. AB - Pulmonary toxicity, as an adverse effect of methotrexate (MTX) therapy, is uncommon in psoriatics. This report concerns a patient with psoriatic arthritis who developed fatal pneumonitis with a histopathological pattern of the organizing stage of diffuse alveolar damage and who was receiving MTX at a dose of 15 mg weekly for 1 month. The patient died despite the immediate withdrawal of MTX, the administration of corticosteroids, and adequate supportive care. Since MTX pneumonitis is a potentially fatal complication, new pulmonary symptoms, even in patients on low-dose MTX treatment, should be appropriately investigated. PMID- 15132234 TI - Surveillance of completed suicide in the Department of the Navy. AB - In 1999, the Department of the Navy (DoN) began a suicide surveillance program using the DoN Suicide Incident Report (DONSIR) to collect data on completed suicides in the Navy and Marine Corps. The DONSIR significantly advances the capability of DoN to track and analyze data on completed suicides over time. The long-term goal of this program is to improve suicide prevention by identifying and modifying military-specific risk factors. A DONSIR has been completed on 98% of the 200 DoN suicides that occurred from 1999 to 2001. Most DoN suicides occurred outside the military work environment and involved the use of a firearm. Most decedents were men, had experienced a recent relationship problem, and did not use any military support services in the 30 days before suicide. Results suggest that the promotion of support services may improve the effectiveness of DoN suicide prevention. PMID- 15132235 TI - Physiological arousal among women veterans with and without posttraumatic stress disorder. AB - The purpose of this study was to assess baseline physiological arousal in women veterans with posttraumatic stress disorder (PTSD) in a nonresearch setting. Heart rate, blood pressure, sublingual temperature, and weight were obtained from a retrospective chart review of the medical records of 92 women veterans with and without a diagnosis of PTSD who were seen in an outpatient Veterans Affairs medical center. Women veterans with PTSD had statistically significantly higher mean baseline heart rates compared with women veterans without PTSD. The two groups did not differ statistically in blood pressure measures, sublingual temperature, or body mass index. Based on our analyses, this difference is not likely to be an artifact of age, race, body mass index, smoking status, or medication. The mean resting heart rate of women with PTSD was 83.9 beats per minute; it was 77.5 beats per minute in those without PTSD. This elevation in heart rate among women veterans with PTSD suggests an increase in baseline physiological arousal compared with women veterans without PTSD. Faster resting heart rate has been shown to be associated with a higher risk of developing hypertension and a greater incidence of cardiovascular morbidity and mortality in non-PTSD samples. Further research is needed to determine the physiological effects of PTSD in women. PMID- 15132236 TI - Antipersonnel mine injuries in Southern Croatia. AB - OBJECTIVES: In this retrospective study, antipersonnel mine casualties in Southern Croatia from 1991 to 1995 are analyzed and treatment options are discussed. METHODS: Mechanism, degree of injury according to Abbreviated Injury Scale (AIS) and Injury Severity Score (ISS), as well as surgical treatment were analyzed. RESULTS: Of a 2,693 war trauma population, 422 (15.67%) patients sustained antipersonnel mine injuries, 241 (57.11%) from pressure mines and 181 (42.89%) from fragmentation mines. Military personnel were injured in 329 cases, civilians in 60 cases, and children in 33 cases. AIS was 3.01 +/- 0.56 and ISS was 17.92 +/- 6.59. Of 39 fatalities (9.24%) with a mean age of 27.98 +/- 1.70 years, 34 were soldiers, 4 were civilians, and 1 was a child. AIS was 5.35 +/- 0.39 and ISS was 54.94 +/- 2.36. CONCLUSION: Fatalities and morbidity arising from antipersonnel mines can be reduced by the provision of appropriate surgical and evacuation facilities at the actual battlefield. PMID- 15132237 TI - Injuries from Antitank mines in Southern Croatia. AB - OBJECTIVE: Antitank mines inflict devastating injuries that are usually fatal. The objective of this retrospective study was to analyze antitank mine casualties in South Croatia during the period from 1991 to 1995. METHODS: Mechanism, degree of injury according to Abbreviated Injury Scale and Injury Severity Score, as well as surgical treatment were analyzed. FINDINGS: Of 464 mine victims, 42 (9.0%) patients sustained antitank mine injuries, and 12 of these were fatal (29%). Abbreviated Injury Scale of the antitank mine injuries was 5.3 +/- 10.6. Military personnel were injured in 29 cases, and civilians were injured in 13 cases. CONCLUSION: Although injuries from antitank mines were ravaging, and frequently fatal, a significant number of patients survived. PMID- 15132238 TI - Homocysteine, vitamins, and prevention of vascular disease. AB - Within the past four decades, the efforts of investigators worldwide have established the amino acid homocysteine as an important factor in arteriosclerosis and diseases of aging. After its discovery in 1932, homocysteine was demonstrated to be an important intermediate in the metabolism of amino acids. However, little was known about the broader biomedical significance of homocysteine until 1962, when children with mental retardation, accelerated growth, dislocated ocular lenses, and frequent vascular thrombosis were found to excrete homocysteine in the urine. My study of two patients with homocystinuria caused by different inherited enzymatic disorders in 1968 disclosed advanced widespread arteriosclerotic plaques in both cases. This discovery led to the conclusion that homocysteine causes vascular disease by a direct effect on the cells and tissues of the arteries. This interpretation suggests that homocysteine is important in the pathogenesis of arteriosclerosis in persons with hereditary, dietary, environmental, hormonal, metabolic, and other factors predisposing them to hyperhomocysteinemia. Within the past decade, many major clinical and epidemiological studies have proven that hyperhomocysteinemia is a potent independent risk factor for vascular disease. According to the homocysteine theory of arteriosclerosis, insufficient dietary intake of the B vitamins, folic acid and pyridoxine, caused by losses of these nutrients during processing of foods, leads to elevation of blood homocysteine and vascular disease in the general population. The dramatic decline in cardiovascular mortality since the 1960s in the United States is attributed to fortification of the food supply by synthetic pyridoxine and folic acid. The recent Swiss Heart Study showed that B vitamins slowed restenosis in patients with coronary arteriosclerosis treated with angioplasty. Currently, more than 20 prospective, worldwide, interventional trials involving at least 100,000 participants are examining whether lowering plasma homocysteine levels with supplemental B vitamins will prevent mortality and morbidity from arteriosclerotic vascular disease. PMID- 15132239 TI - Identification of Francisella tularensis using real-time fluorescence polymerase chain reaction. AB - A Francisella tularensis-specific, TaqMan probe-based, real-time fluorescence polymerase chain reaction (PCR) assay required approximately 60 minutes and consistently achieved a sensitivity of < or = 10 fg of F. tularensis genomic DNA (five genome equivalents). Specificity testing against a genomic DNA cross reaction panel comprised of 22 bacterial organisms representing closely related species, diverse genera, and human genomic DNA resulted in no false positives of significance. The assay was conducted on a field-deployable thermocycler, the R.A.P.I.D. ("Ruggedized" Advanced Pathogen Identification Device), a microbial identification system that can provide rapid and accurate identification F. tularensis. PMID- 15132240 TI - Late traumatic flap displacement after laser in situ keratomileuisis. AB - Laser in situ keratomileusis (LASIK) has become the community standard in corneal refractive surgery and is being performed by surgeons in the Army, Navy, and Air Force. LASIK differs from photorefractive keratectomy (PRK) in that a partial thickness corneal flap is created in the LASIK procedure before removing a microscopic amount of corneal tissue, whereas no flap creation is required in PRK. The benefits of LASIK include minimal discomfort after surgery, as well as a much faster return of visual function. PRK involves a surface ablation and therefore heals differently, involving more discomfort and a slower return of functional vision. LASIK flap integrity is a concern to anyone undergoing the procedure, as well as for those making recommendations on the best form of refractive surgery for military personnel. A case report and a review of the literature are presented on the identification and management of LASIK flap trauma. PMID- 15132241 TI - Upper respiratory tract infections (URI). PMID- 15132242 TI - HIPAA one year later: effects and pitfalls. PMID- 15132243 TI - Depression in children and adolescents: the latest evidence-based psychopharmacological treatments. PMID- 15132244 TI - Family member response to violent infant kidnapping. AB - All family members interviewed suffered from chronic posttraumatic stress disorder 5 or more years after an infant kidnapping. Psychiatric nurses should routinely assess for secondary victimization related to any type of traumatic stressor. Nurses should refer parents and families to materials available from the National Center for Missing and Exploited Children (NCMEC) (http://www.missingkids.com/). PMID- 15132245 TI - Screening young people in the juvenile justice system for behavioral & substance abuse disorders. AB - There is a wide variation in viewpoints regarding screening youth in the juvenile justice system for behavioral and substance abuse disorders. No consensus exists within and among local, state, and federal juvenile justice officials regarding the necessity of screening youth for behavioral and substance abuse disorders. Local, state, and federal juvenile justice officials voiced concern regarding screening young people for behavioral and substance abuse disorders without having available funding or facilities for diagnosis and treatment. PMID- 15132246 TI - Fallen apples. AB - Children today often have to cope with living in fractured family systems in which multigenerational dysfunction and abuse is passed on to them. As a result, they often make decisions that lead them into unsafe, and possibly disastrous situations. Some children and adolescents cannot see beyond today. They are prone to giving up because they feel nothing can change. Growing up in physically violent homes can create children who are unable to control their own violent rages. While hospitalized, they often seek the struggle and emotional release that accompanies physical management. Physical management allows the children to be out of control and safe at the same time. PMID- 15132247 TI - Cerebral palsy: defining the problem. AB - The term "cerebral palsy" has had varied meanings over the past century. This article presents the current consensus definition that best captures the core elements of this heterogeneous entity. Elements comprising the components of the consensus definition are elaborated upon to provide clarity. Areas of remaining lack of consensus, mostly reflecting the timing of diagnosis and the exclusion of various etiologic entities are highlighted. PMID- 15132248 TI - The World Health Organization International Classification of Functioning, Disability, and Health: a model to guide clinical thinking, practice and research in the field of cerebral palsy. AB - The way we think about health and disease determines to a considerable extent what we do and say in our clinical encounters with patients. The recent publication and promotion of the World Health Organization's International Classification of Function, Health, and Disability (known as the ICF) represents an exciting new way to consider health and disease. In the context of children and youth with cerebral palsy, this model offers many heretofore ignored "point of entry" for counselling and intervention with these conditions. This model also provides many possibilities to explore research questions with a fresh approach. This article outlines the ICF model and discusses these opportunities. PMID- 15132249 TI - New directions in the outcome evaluation of children with cerebral palsy. AB - This article addresses newly emerging concepts regarding the examination of outcomes in children with cerebral palsy (CP). Three important factors have prompted a change toward a broader approach to outcome measurement: (1) the recent endorsement of the World Health Organization classification scheme of functioning, disability, and health; (2) recognition of the importance of the role of child and environmental factors in modifying outcome; and (3) appreciation of the challenges faced at key transitions during development. Comparison of the literature on outcomes in children with CP published in 2002 with studies published 10 years earlier demonstrates an increased number of studies and a wider range of outcome measures used to characterize the functioning of these children. A better understanding of the extent and nature of the outcomes and their determinants is critical to guide clinical decision making and program planning to maximize community integration, social participation, and overall well-being of the child and family. PMID- 15132250 TI - Cerebral palsy registries. AB - Cerebral palsy (CP) registers appear to be appropriate tools for answering questions regarding the prevalence and characteristics of this common childhood disability. Registers are population databases issuing from multiple sources, relying on a clear definition and inclusion and exclusion criteria of CP, and requiring a mix of skills with the collaboration of obstetricians, pediatricians, and epidemiologists. In Europe alone there are 18 different CP registers or population data collections on CP, and collaborative research efforts exist through a European network. Data collection on CP has also been done in Australia (register), the United States (surveys), and Canada (register). Beside monitoring trends, other public health contributions of CP registers might be to reduce the frequency of CP and to improve the quality of life of children with CP. CP registers are useful to clinicians by enabling them to identify subgroups of children requiring specific etiologic investigations, and also to provide more accurate information to the parents of children with CP. PMID- 15132251 TI - The "Bermuda triangle" of neonatal neurology: cerebral palsy, neonatal encephalopathy, and intrapartum asphyxia. AB - The terms "cerebral palsy," "neonatal encephalopathy," and "intrapartum asphyxia" are frequently used in pediatric neurology. This article presents concise, verifiable definitions for each of these entities based on our current understanding and formulates the nature of the interrelationships between them. The aim is to provide a level of clarity that will enhance diagnostic and pathogenetic precision and minimize conceptual misunderstanding. This should aid future therapeutic and research efforts in this important area. PMID- 15132252 TI - Animal models of hypoxic-ischemic brain damage in the newborn. AB - Controversy continues over which animal model to use as a reflection of human disease states. With respect to perinatal brain disorders, scientists must contend with a disease in evolution. In that regard, the perinatal brain is at risk during a time of extremely rapid development and maturation, involving processes that are required for normal growth. Interfering with these processes, as part of therapeutic intervention must be efficacious and safe. To date, numerous models have provided tremendous information regarding the pathophysiology of brain damage to term and preterm infants. Our challenges will continue to be in identifying those infants at greatest risk for permanent injury, and adapting therapies that provide more benefit than harm. Using animal models to conduct these studies will bring us closer to that goal. PMID- 15132253 TI - Evaluation of the child with cerebral palsy. AB - Cerebral palsy (CP) is a common problem, occurring in about 2 to 2.5 per 1000 live births. The diagnosis of CP is based upon a history of abnormal motor development that is not progressive coupled with an examination (e.g. hypertonicity, increased reflexes, clonus) "placing" the lesion in the brain. In order to establish that a brain abnormality exists in children with CP that may, in turn, suggest an etiology and prognosis, neuroimaging is recommended with magnetic resonance imaging preferred to computed tomography. Metabolic and genetic studies should be obtained if there are atypical features in the history or on the examination. Detection of a brain malformation in a child with CP might suggest an underlying genetic or metabolic etiology. As cerebral infarction is high in children with hemiplegic CP, diagnostic testing for coagulation disorders should be considered. However, there is insufficient evidence at present to be precise as to what studies should be ordered. An electroencephalogram is not recommended unless there are features suggestive of epilepsy or a specific epileptic syndrome. As children with CP may have associated deficits of mental retardation, ophthalmologic and hearing impairments, speech and language disorders and oral-motor dysfunction, screening for these conditions should be part of the initial assessment. PMID- 15132254 TI - Management of spasticity in children with cerebral palsy. AB - As one component of the upper motor neuron syndrome, spasticity can have a significant functional impact on the child with cerebral palsy. Treatment planning requires the determination that excess tone interferes with some aspect of function, comfort, or care, and takes into consideration carefully devised goals that meet the needs of the patient and the caregiver. Treatment options include physical therapy, oral medications, chemodenervation with botulinum toxin or phenol, rhizotomy, intrathecal baclofen, and orthopedic surgery. The uses and limitations of each is discussed, and evidence for efficacy in cerebral palsy is reviewed. PMID- 15132255 TI - Recent advances in physical and occupational therapy for children with cerebral palsy. AB - Recent research and conceptual frameworks that emphasize family centered care and participation in family, school, and community life have contributed to advances in provision of health care by physical therapists and occupational therapists to children with cerebral palsy (CP) and their families. This article provides a contemporary perspective on physical and occupational therapy for children with CP. Topics include conceptual frameworks, early identification, prognosis for gross motor function, implications of impairment in motor control and muscle performance, and physical fitness and secondary prevention. Case scenarios and examples illustrate how current knowledge and research is applied to decision making within the context of the individual child and family. PMID- 15132256 TI - Family-centered service for children with cerebral palsy and their families: a review of the literature. AB - Pediatric neurologists and other specialists working with children with cerebral palsy or other disabilities play important roles in providing services in ways that make a difference in the lives of these children and their families. This article defines family-centered service and outlines a conceptual framework of the premises and principles underlying this approach to service delivery. Research evidence supporting family-centered service is presented for child, parent/family and service system outcomes, with a focus on community-based rehabilitation or health care services. The research evidence shows strong support for family-centered service in promoting the psychosocial well-being of children and their parents and in leading to increased satisfaction with services. There is less evidence for other outcomes. It is recommended that all service providers assess their beliefs and behaviors about family-centered service. PMID- 15132257 TI - ICF: health vs disease. PMID- 15132258 TI - Participation of children with cerebral palsy is influenced by where they live. AB - The study aimed to determine whether degree of participation of children with cerebral palsy (CP) is influenced by where they live, as predicted by the social model of disability. Ninety-two per cent children with CP resident in Northern England and born 1991-1996 were entered into the study. Participation was measured by the Lifestyle Assessment Score and its six component domain scores. Regression analysis was used to investigate variations in participation. There were 443 children (265 male, 178 female; mean age 4 years 8 months [SD 1 year 1 month] at time of assessment) in the study. In the regression analysis the following factors remained significant with regard to level of participation: type of CP (167 with hemiplegia, and of those remaining 240 with bilateral spasticity); intellectual impairment (105 with IQ<50, 113 with IQ 50 to 70, and 225 with IQ>70); presence of seizures (115 with active epilepsy); walking disability (114 unable to walk, 81 restricted and needing aids, 186 restricted but unaided, 62 unrestricted); communication problems (61 no formal communication, 51 use alternative formal methods, 126 some delay or difficulty, 205 no communication problems). After adjustment for these factors, there were significant variations with regard to level of participation in the Lifestyle Assessment Score by district of residence. The magnitude of these variations in Lifestyle Assessment Score between districts is similar to that accounted for by severe intellectual impairment. Similar models were obtained for four of the six domain scores. For one of these four, restriction of social interaction, the significant variation between districts was minimally influenced by the underlying type of CP, walking ability, or presence of seizures. Higher levels of participation among children with CP are associated with residence in certain districts. This is not attributable to variations in case-mix or functional capacity of the children. Participation of children with disability is partly a product of their environment. PMID- 15132259 TI - Identification and description of environmental factors that influence participation of children with cerebral palsy. AB - Physical, social, and attitudinal environment may restrict participation in children with cerebral palsy (CP). Here we discuss existing/possible approaches in order to identify and describe this environment. We used a critical review of evidence from the World Health Organization Literature Review on Environmental Factors; a search of electronic databases; and talked to specialists in order to find unpublished papers and 'grey' literature. Both children with disabilities and their parents identified a range of barrier and facilitator factors. These included psychosocial pressures (family, school), financial difficulties, and inadequate public services. Observational studies suggest that building structure, loss of income, and provision of specific equipment have a direct impact on levels of child participation. Some available instruments attempt to capture environmental factors by client survey or objective measurement; most relate to adult contexts, but there are a few child-specific instruments for surveying attitudes of children to peers with disabilities and for observation of the school environment. Defining and measuring potential environmental determinants of participation for children with CP needs further development; and here we propose how this might be done. PMID- 15132260 TI - Bodily pain and health-related quality of life in children with cerebral palsy. AB - Pain frequency and its association with participation, function, and health related quality of life were evaluated in a sample of 198 children (58% male; mean age 10 years 7 months, SD 3 years 11 months; range 5 to 18 years) with moderate to severe cerebral palsy (CP). Gross Motor Functional Classification System (GMFCS) levels were III to V (23% level III, 23% level IV, 31% level V without a gastrostomy tube, and 23% were level V with a gastrostomy tube). Assessment was carried out using the parent-report Child Health Questionnaire (CHQ) and questions assessing health, medical utilization, and medication use. Pain frequency ranged from 'none' to 'every day'. Eleven percent of parents reported pain almost every day (z score=-0.38, p<0.001). Pain was related to severity of motor impairment and the presence of a gastrostomy; parents of the most severely affected children reported the highest pain frequency (p=0.05). Pain was correlated with school days missed (p=0.03) and days in bed (p=0.01). Children taking gastrointestinal medications were reported to have greater pain frequency (p<0.001). The Parental Impact--Emotional subscale of the CHQ was correlated with reported pain (r=0.38, p<0.001). Pain was frequent in children with moderate or severe CP. Pain was more prevalent with more severe impairment and was associated with educational and social consequences. PMID- 15132261 TI - Gross Motor Function Classification System and outcome tools for assessing ambulatory cerebral palsy: a multicenter study. AB - The relationships between different levels of severity of ambulatory cerebral palsy, defined by the Gross Motor Function Classification System (GMFCS), and several pediatric outcome instruments were examined. Data from the Gross Motor Function Measure (GMFM), Pediatric Orthopaedic Data Collection Instrument (PODCI), temporal-spatial gait parameters, and oxygen cost were collected from six sites. The sample size for each assessment tool ranged from 226 to 1047 participants. There were significant differences among GMFCS levels I, II, and III for many of the outcome tools assessed in this study. Strong correlations were seen between GMFCS level and each of the GMFM sections D and E scores, the PODCI measures of Transfer and Mobility, and Sports and Physical Function, Gait Velocity, and Oxygen Cost. Correlations among tools demonstrated that the GMFM sections D and E scores correlated with the largest number of other tools. Logistic regression showed GMFM section E score to be a significant predictor of GMFCS level. GMFM section E score can be used to predict GMFCS level relatively accurately (76.6%). Study data indicate that the assessed outcome tools can distinguish between children with different GMFCS levels. This study establishes justification for using the GMFCS as a classification system in clinical studies. PMID- 15132262 TI - Comparison of direct and indirect measures of walking energy expenditure in children with hemiplegic cerebral palsy. AB - The purpose of this study was to investigate the use of the heartrate (HR) version of the energy expenditure index (EEIHR) as a proxy for measurement of walking oxygen consumption (VO2) in children with cerebral palsy (CP). Thirteen children (eight males, five females; mean age 11 years 2 months [SD 3 years], age range 6 to 15 years) with hemiplegic CP, participated in this study. The study was conducted over three sessions. During session 1, participants were familiarized with testing procedures and given 5 minutes of treadmill walking practice. In session 2, participants completed three 5-minute walking bouts on the treadmill at 0.67m x s(-1) to familiarize themselves with treadmill locomotion. During the final session participants walked at 0.67, 0.89, and 1.12m x s(-1) for 5 minutes while gross oxygen consumption (gross VO2; walking VO2/speed), net VO2 ([walking VO2-resting VO2]/speed), and EEIHR ([walking HR resting HR]/speed) were measured during the last 2 minutes of each bout. Correlational analyses indicated no relationship (p>0.05) between measures of gross VO2 and EEIHR at each speed. Although no association was evident between net VO2 and EEIHR at 0.67 and 0.89m x s(-1), a moderate relationship (r=0.64; p<0.05) was present between these variables at 1.12m x s(-1). Examination of individual data revealed that most participants displayed an unmatched pattern of response between net VO2 and EEIHR. Our results suggest that caution should be applied when using EEIHR to estimate walking energy expenditure in children with CP. PMID- 15132263 TI - Control of prehension in hemiparetic cerebral palsy: similarities and differences between the ipsi- and contra-lesional sides of the body. AB - This study aimed at broadening our insight into ipsi- and contra-lesional control of prehension after unilateral brain damage. Six male adults with hemiparetic cerebral palsy (mean age 17 years 3 months, SD 15 months) performed unrestricted grasping of discs that differed in size (40, 60, and 80mm in diameter) and which were placed at different distances from the participants (150mm and 30mm). A precalibrated Optotrak 3020 system was used for recording motion. Kinematics of the transport and grasp component, and hand orientation at the moment of grasping, were determined. A marker on the wrist was used to calculate the kinematics of the transport component. The distance between the markers on the index finger and thumb was used for the calculation of the grasp component. Kinematic variables of the transport and grasp component were remarkably similar between both sides of the body. However, with the contra-lesional side, more time was spent in contact with the object before it was lifted, and movements were performed less fluently compared with the ipsi-lesional side. Maximum grasp aperture was attained very late during reaching. For final hand orientation, results showed a large standard deviation both within and between participants. These findings show that, despite the ostensible awkward prehension movements of the contra-lesional side (slowness and decreased fluency), similarities in the kinematics between both sides of the body are present, which may be indicative of intact central control of the movement. The results of variant final hand orientation, combined with the relative late occurrence of peak aperture, suggest that these participants encounter difficulties with forward planning and may use a step-by-step control strategy. PMID- 15132264 TI - Visual search in school-aged children with unilateral brain lesions. AB - In this preliminary study, visual search for targets within and beyond the initial field of view was investigated in seven school-aged children (five females, two males; mean age at testing 8 years 10 months, SD 1 year 3 months; range 6 to 10 years) with various acquired, postnatal, focal brain injuries (haematoma, haemorrhage, meningioma, neuroblastoma, and cerebral abscess) in anterior or posterior sites of the left or right hemisphere, and seven control children (matched for age and sex) were also studied. All participants attended mainstream primary schools. The children with lesions underwent surgery after diagnosis (mean age at diagnosis 5 years 4 months, SD 2 years 7 months). Group results indicated that for the overall scores on three psychometric tests of visuospatial and fine motor abilities (Southern California Figure Ground Perception Test, Visual Organization Test, and Visual-Motor Integration Test), no difference between the children with left and right lesions was present. However, children with lesions in the right hemisphere, and not in the left hemisphere, took significantly more time than the controls to locate visual targets presented within and beyond the field of view. Examination of individual data suggested that, in accordance with brain imaging research, right-sided anterior cerebral lesions sustained in early childhood might have an enduring detrimental effect on voluntary visual search performance during development. This persistent effect of early brain injury might imply that developmental plasticity of the brain does not apply to certain specific functions of particular areas of the right hemisphere. PMID- 15132265 TI - Cognitive development in Silver-Russell syndrome: a sibling-controlled study. AB - This study examined cognitive development in Silver-Russell syndrome (SRS), a condition with intrauterine growth retardation, persisting short stature, and specific stigmata. Neuropsychological function and cognitive abilities were assessed in a sample of 36 children with SRS (21 males, 15 females; mean age 7 years 6 months, SD 2 years 8 months; age range 3 to 12 years) and 25 normally developing siblings (mean age 8 years 6 months, SD 2 years 7 months; age range 3 to 12 years) using the Kaufman Assessment Battery for Children. Special measures were taken to control for confounding factors and sample bias. Mean overall IQ score in the total SRS sample (95.7, SD 10.63), as measured by the Mental Processing Composite Scale, was significantly reduced (p=0.021) compared with test norms (IQ 100, SD 15), indicating a moderate cognitive impairment. Subscale analysis revealed some specific deficiencies. However, these cannot be attributed to an established category of specific learning disorder. The mean score in the Achievement Scale (91.25, SD 14.92), which is more sensitive to educational influences, showed stronger deficits (p=0.001). The sibling control group achieved a slightly better mean IQ score (104.20, SD 12.32) than test norms (p=0.10). Direct analysis of paired differences between the subsample of children with SRS and a sibling among the control group (n=25) revealed a significant mean difference of 8.08 IQ points (p=0.011). Risk factor analysis revealed that cognitive development is not associated with birth length (p=0.404), birthweight (p=0.820), growth hormone therapy (p=0.810), phenotypic severity (p=0.828), or sex (p=0.880). Two children with maternal uniparental disomy for the entire chromosome 7 had markedly lower IQ scores (81 and 84 respectively). In contrast to the few previous findings, children with SRS show only moderate, but significant, impairments in cognitive outcome, which are more striking in our sample when compared with siblings than with test norms. PMID- 15132266 TI - Photosensitivity and epilepsy: a follow-up study. AB - To understand the evolution of photosensitivity and to evaluate if its disappearance is related to the response to anticonvulsant therapy, we performed a long-term study of 42 patients (17 males, 25 females; age at onset 6 years 9 months, SD 5 years 2 months, range 5 years to 12 years 1 month) who had electroencephalography (EEG) evidence of photosensitive epilepsy. Of the patients, 36 were treated with valproate (VPA) monotherapy and four received VPA in combination with other antiepileptic drugs (AEDs), which were carbamazepine and lamotrigine. Two patients were given no drugs, but treated with stimuli avoidance. All patients were investigated with EEG by using intermittent photic stimulation. The photoparoxysmal response indicated the presence of photosensitivity. At the end of follow-up, the photoparoxysmal response had disappeared in 25 patients. Thirty-three patients became seizure-free. Our study confirms that photosensitive epilepsy has a good prognosis for seizure control that is independent of the persistence or disappearance of photosensitivity. PMID- 15132267 TI - Efficacy of bracing the lower limbs and ambulation training in children with myelomeningocele. PMID- 15132268 TI - Opioid peptides and dipeptidyl peptidase in autism. PMID- 15132269 TI - Does cerebral perfusion pressure influence outcome in children with tuberculous meningitis? PMID- 15132270 TI - Behavior management for the pediatric dental patient. PMID- 15132271 TI - Challenges of managing child behavior in the 21st century dental setting. AB - This paper discussed factors influencing behavior management of the child dental patient. Pediatric dentists are affected by changes in: (1) society; (2) marketing and media; (3) communications and technology; and (4) parenting practices. Behavior of pediatric patients reflects fewer boundaries, less discipline and self-control, and lowered behavioral expectations by parents and contemporary culture. The insurance industry, regulatory bodies, legal system, dental staff, and pediatric dentist education are other influences on behavior management. Responses of the American Academy of Pediatric Dentistry (AAPD), which could support the pediatric dentist in the changing environment, include: (1) research; (2) continuing education for staff and dentists; (3) development of Internet accessible materials for the public; (4) legislative activity; (5) partnering with pediatric medicine to develop new behavior management strategies; (6) establishment of an AAPD Council on Child Behavior; and (7) ongoing critical reassessment of behavior issues by the AAPD. PMID- 15132272 TI - The child's voice: understanding the contexts of children and families today. AB - Children and families live and grow in a different social context than 15 years ago. The purpose of this report was to explore contemporary themes for improved contextual understanding of children and families today and their relationship to providing optimal pediatric dental care. Twelve themes were discussed using research studies and clinical expertise from the viewpoints of a pediatric psychologist and pediatrician. It was concluded that enhanced communication and partnership building improves comprehension and compliance with dental treatment. Furthermore, diagnosing the child and family within the immediate local context is central to developing and accomplishing an effective dental treatment plan. PMID- 15132273 TI - The changing nature of parenting in America. AB - The purpose of this paper was to discuss how parenting has changed in recent decades. The topics discussed include: (1) increased focus on parenting in today's culture; (2) perception of pediatric dentists regarding parenting changes; (3) relative importance of parenting in children's development; (4) family/parenting trends; and (5) the role of stress in the changing nature of parenting. PMID- 15132274 TI - Managing pediatric dental patients: issues raised by the law and changing views of proper child care. AB - The purpose of this paper was to examine legal issues regarding the management of pediatric dental patients and changing views of proper child care. Standards of care in pediatric dentistry are not static. They change in response to research, patterns of reimbursement, patient and parental expectations of reasonable care, and consensus among practitioners. The law pertaining to accountability for pediatric dental patient treatment largely reflects standards of care established by the pediatric dentistry profession. However, the law can also reflect changes in public expectations of reasonable care that can effectively outrun the discipline's efforts to reflect new knowledge or changing public concerns. A major impetus for considering the care of children in all settings has been the increasing recognition of suboptimal children's care, as well as concerns that children have either been abused or neglected in a number of settings. Too often, practices towards children have been untested and based only on the assumption that what is done is "for the child's own good." Pediatric dentists can respond to changing standards of reasonable care for pediatric dental patients, as expressed in legal decisions. They can also usefully consider how attention to child maltreatment has sensitized parents to be better consumers of services on their children's behalf. Rather than reacting only to public pressures for better means of behavior management, the challenge is to exceed expectations via new research and thoughtful anticipation of improvements that can be made. PMID- 15132275 TI - Pharmacological management of the pediatric dental patient. AB - Pharmacological management of the pediatric dental patient is considered a subcategory of a broader collection of professional mediated activities known as behavior management techniques. Pharmacological techniques are generally divided into either: (1) various levels of sedation; or (2) general anesthesia. Pharmacological techniques are not universally offered by practicing dentists for a host of reasons including, but not limited to: (1) variation in practitioner training and philosophy; (2) state rules and regulations; (3) cost and reimbursement; and (4) safety issues. PMID- 15132276 TI - Issues regarding insurance and other third-party reimbursement for behavioral management procedures. AB - Issues related to reimbursement for time and effort expended in providing behavior management services for children often result in confusion and frustration for the parent and practitioner. Multiple medical and dental insurance plans within a state or region frequently lack a common set of definitions that are fundamental in interpreting and applying contract language. Ambiguities and inconsistencies in state and federal programs and regulatory structures, as well as budget concerns, only exacerbate the problem. This paper reviews issues of third-party payment associated with dental care for children requiring behavioral management. An environmental assessment of the health care system identifies individual and societal challenges. A model that identifies stakeholders involved in third-party reimbursement is discussed, noting the incentives faced by each stakeholder and expected behaviors based on those incentives. Strategies for effecting change based on a thorough understanding of each stakeholders' concerns are discussed. PMID- 15132277 TI - Survey of behavior management teaching in predoctoral pediatric dentistry programs. AB - PURPOSE: The purpose of this study was to survey directors of predoctoral pediatric dentistry programs regarding the teaching of behavior management techniques. METHODS: Surveys were mailed to all 56 dental schools in the United States. Follow-up mailings were sent to nonrespondents. The survey contained items on program demographics and the program's teaching of communicative and pharmacologic techniques. Information was also obtained on informed consent and parental presence in the operatory. RESULTS: Surveys were returned by 48 schools. Two schools declined to respond because they had not yet accepted or graduated students. The final response rate was 89%. The mean (+/-SD) percentage of total didactic time devoted to behavior management was 12% (+/-6). Communicative techniques were taught as "acceptable" by 96% to 100% of programs, with the exception of the hand-over-mouth exercise (HOME). HOME was taught as "unacceptable" by 62% of programs. Active and passive immobilization of sedated and nonsedated children was taught as "acceptable" by 69% to 85% of programs. Sixty-seven percent to 98% of programs taught that pharmacologic techniques (nitrous oxide, conscious sedation, general anesthesia) are "acceptable." There was little evidence that the teaching of behavior management techniques had changed over the previous 5 years, nor that they were likely to change in the near future. Parental presence in the operatory was common for some procedures, particularly among younger children. CONCLUSIONS: Predoctoral programs teach as acceptable communicative and pharmacologic management techniques, with the exception of HOME. Predoctoral program directors report they are not likely to increase the amount of curricular time devoted to behavior management in the near future. PMID- 15132278 TI - Survey of behavior management teaching in pediatric dentistry advanced education programs. AB - PURPOSE: The purpose of this study was to survey pediatric dentistry advanced education program directors regarding the teaching of behavior management techniques. METHODS: Surveys were mailed to all (65) advanced education programs in the United States. Follow-up mailings were sent to nonrespondents. The survey contained items on program demographics and the program's teaching of communicative and pharmacologic techniques. Information was also obtained on informed consent and parental presence in the operatory. RESULTS: Surveys were returned by 54 programs. Two programs declined to respond because they had not yet accepted or certified residents. The final response rate was 86%. The mean percentage (+/- SD) of total didactic time devoted to behavior management was 13% (+/-9.5). Communicative techniques were taught as "acceptable" by 98% of programs, with the exception of the hand-over-mouth exercise (HOME), which was taught as "unacceptable" by 54% of programs. Active and passive immobilization of sedated and nonsedated children was taught as "acceptable" by 76% to 98% of programs. All programs taught that pharmacologic techniques (nitrous oxide, conscious sedation, general anesthesia) are "acceptable." There was little evidence that the teaching of behavior management techniques had changed over the previous 5 years, nor that it is likely to change in the near future. Parental presence in the operatory was common for some procedures, particularly among younger children. CONCLUSIONS: Most programs do not teach HOME as an acceptable behavior management technique. The amount of curricular time devoted to behavior management is not likely to change appreciably in the near future. PMID- 15132279 TI - A survey of members of the American Academy of Pediatric Dentistry on their use of behavior management techniques. AB - PURPOSE: The objective of this study was to survey members of the American Academy of Pediatric Dentistry (AAPD) regarding their use of behavior management techniques. METHODS: Surveys were mailed to 4,180 members, with a follow-up mailing to nonrespondents 2 months later. The survey contained items on demographic variables and use (current, past, and future) of communicative and pharmacologic techniques. Information was also obtained on informed consent, parental presence in the operatory, and parenting styles. RESULTS: Survey response was 66%. Communicative techniques are widely used, with the exception of the hand-over-mouth exercise (HOME). Immobilization for sedated and nonsedated children and pharmacologic techniques are used by a majority or near majority of respondents. Little change was reported in technique use over time, except that 50% of respondents indicated they use HOME less now than 5 years ago, and 24% plan to use it less over the next 2 to 3 years. Parental presence in the operatory appeared to be a common practice for some procedures and for children with special health care needs. The majority of respondents believed that parenting styles had changed in ways that adversely impacted children's behavior in the dental setting. CONCLUSIONS: Most practitioners have not changed their use of behavior management techniques in recent years, nor do they plan to change their use of them in the near future. HOME was the exception to these trends. PMID- 15132280 TI - Behavior management conference panel I report--Rationale for behavior management techniques in pediatric dentistry. AB - Panel I comprised of pediatric dentists, an attorney, child psychologists, parents, a specialist in early childhood education, and a pediatrician. The purpose of this panel was to discuss: (1) 8 questions that dealt with the appropriateness and effectiveness of current behavior management techniques; (2) the scientific support for those techniques; and (3) the role of the pediatric dentist in managing the difficult child. Issues of cultural diversity, access to care, and parental attitudes toward behavior management were also explored. Nonpediatric dentist members of the panel offered insights into how other health care professionals view the behavior management techniques used by pediatric dentists. The panel sought input from the conference attendees as part of its deliberations. The major recommendations of the panel included: (1) re-evaluate the definitions of child behavior in the dental setting, including definitions of appropriate behavior; (2) develop training in effective communication with parents for pediatric dentists and their staffs; (3) seek further information on the impact that changing parental attitudes towards behavior management techniques may have on the quality and accessibility of treatment; and (4) conduct research in specific areas of behavior management, particularly in communicative techniques. PMID- 15132281 TI - Behavior management conference panel II report--Third-party payer issues. AB - Panel II discussed key issues related to third-party payment for behavior management in pediatric dentistry and responded to a series of questions raised by the Conference Planning Committee. The panel was composed of individuals representing consumers (parents and caretakers), pediatric dentists, other health care providers, and a large dental insurance plan. They were charged with: (1) identifying problems related to third-party coverage and benefits for behavior management; (2) developing recommendations for the American Academy of Pediatric Dentistry; and (3) prioritizing the problems and recommendations. PMID- 15132282 TI - Behavior management conference panel III report-Legal issues associated with managing children's behavior in the dental office. AB - Panel III reviewed the legal issues associated with managing child behavior in the dental setting. The first issue addressed was a review of the recent changes in informed consent for behavior management techniques in pediatric dentistry. Discussions focused on: (1) who should obtain informed consent; (2) who can give informed consent; and (3) what constitutes appropriate documentation. Recent legal actions related to poor outcomes and poor communication using behavior management techniques were identified and discussed. Addressed was the current movement by the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) towards empowering the child-patient to be a co-decision maker at all appointments and its impact on decisions in the dental office about approaches to behavior management. Finally, liability issues accompanying increased use of sedation in the dental office were identified and discussed. The recommendations of the panel were: (1) consideration should be given to changing the language in the current American Academy of Pediatric Dentistry (AAPD) guidelines, so written consent is not required for any procedure; (2) the AAPD should create language for use in creating sample consent forms (suggestive, not prescriptive); (3) guidelines recommending the possibility of treatment deferral, when appropriate, should be strengthened; and (4) the AAPD should work with advanced education program directors to standardize the quality and number of educational experiences in the use of conscious sedation. PMID- 15132283 TI - Behavior management conference panel IV report--Educational issues. AB - Panel IV convened to address whether and how dental education curricula on behavior management needs to adapt to the changing needs and expectations of families and patients. There was a general consensus that behavior management should have a greater focus in dental education. Predoctoral and postdoctoral students may be expected to have different levels of exposure to behavior management concepts and techniques, but communications training should be a greater emphasis throughout dental education. Panel members specifically addressed behavior management techniques, including immobilization, restraint, sedation, and general anesthesia. Major recommendations from the panel included: 1) promote more research in behavior management; 2) update the AAPD guidelines on behavior management; 3) develop competencies/proficiencies for training programs and defining standardized outcomes; 4) introduce an AAPD task force to examine sedation issues that pertain to pediatric dental education; 5) explore new teaching methods of behavior management strategies and techniques; 6) develop a public awareness campaign to educate the public regarding what pediatric dentists do best. PMID- 15132284 TI - Can we afford a further increase in life expectancy? PMID- 15132285 TI - Risk factors and health determinants in older Italians. AB - BACKGROUND AND AIMS: According to a WHO estimate, 50% of the total burden of diseases in men and 25% in women for the EURO-A group (which includes the highly developed countries of the WHO European Region) are attributable to unhealthy life-styles. The aim of the present study was to analyze anthropometric, biochemical and behavioral risk factors using data from the Italian Longitudinal Study on Aging (ILSA), a population-based study of older Italians initiated in 1992 to study those chronic conditions and attendant risk factors that contribute most substantially to morbidity, disability and mortality. METHODS: In this descriptive study, we calculated the means and distribution of risk factors in a sample of 5632 Italian subjects aged 65-84 by gender, age class and geographic area, and identified the proportion of these subgroups at higher risk. Analyses include data from the first (1992-93) and second (1995-96) examinations of this cohort. RESULTS: Over 64% of older Italian participants were overweight in 1992. More than 70% had blood pressure in the borderline or definitely hypertensive range, and almost one-third had hyperlipidemic serum cholesterol levels, including subjects receiving treatment for these conditions. Based on Body Mass Index, more women than men were obese (27 vs 15%) and more women than men were hyperlipidemic in all age classes (38 vs 23%). Approximately 13% of the sample had glucose levels exceeding the recommended 126 mg/dL. Men were found to consume on average about 41 grams of alcohol daily and women 17 g/d. Lastly, we found that approximately 20% of men and 8% of women were smokers in 1992 but that smoking tended to diminish with age. CONCLUSIONS: As the Italian population rapidly ages, the burden of disease and disability is increasing, necessitating more focused, immediate and effective prevention programs. We have identified a number of critical concerns ripe for intervention. The results of this study can better focus such efforts and help guide long-term health planning and policy. PMID- 15132286 TI - Management of chronic obstructive pulmonary disease in the elderly. AB - Chronic obstructive pulmonary disease (COPD), a leading cause of death and disability in the elderly, is frequently unrecognized or misinterpreted as heart disease. Comorbidity plays a primary role, both as a determinant of health status and as a prognostic marker in older populations with COPD. Multidimensional assessment tailored to the distinctive needs of respiratory patients and thus including selected respiratory function indexes, is mandatory for proper staging COPD and monitoring of its course and response to therapy. In stable COPD, a mix of pharmacological and non-pharmacological measures may improve health, but only by stopping smoking and, in the event of respiratory insufficiency, applying continuous oxygen therapy can the progression of the disease be delayed and life expectancy prolonged. In exacerbated COPD, age per se is a negative prognostic marker and, while many very old patients can successfully recover, they will experience some decline in personal independence. Thus, older patients with COPD should ideally be the object of a continuum of care throughout all the stages of their disease, in order to minimize the decline in personal independence and worsening health. In this perspective, COPD patients qualify as optimal candidates for dedicated programs of continuous geriatric care. PMID- 15132287 TI - Mortality and readmission of the elderly one year after hospitalization for pneumonia. AB - BACKGROUND AND AIMS: Pneumonia, which is common among the elderly, is associated with untoward consequences. We sought, therefore, to describe the incidence of death and readmission, and to determine predictors of these variables during the year subsequent to index hospitalization. METHODS: This study involved the follow up of 153 patients surviving an index hospitalization for pneumonia. Death and readmission were documented, and the relationship of selected variables with these outcomes was determined. RESULTS: Ninety-six (62.6%) of the patients had died or were readmitted. Only a count of comorbidities was correlated significantly with death, readmission, and either death or readmission. Using regression analysis, death alone was predicted by multiple variables. Grip strength and comorbidity counts correctly classified 75.2% of patients relative to that outcome. CONCLUSIONS: Untoward outcomes are likely among patients surviving acute hospitalization for pneumonia. These outcomes are related to variables that can be targeted in secondary prevention efforts. PMID- 15132288 TI - Effects of chronic airway disease on health status of geriatric patients. AB - BACKGROUND AND AIMS: The impact of chronic airway disease on the health status of elderly patients is only to some extent explained by indexes of airflow limitation. The present study was designed to assess to what extent: 1) asthma, chronic obstructive pulmonary disease (COPD) and chronic bronchitis with normal FEV1 (simple bronchitis) differ in their impact on health status; 2) health status depends upon non-respiratory factors. METHODS: A total of 1601 outpatients over 65-198 with asthma, 228 with COPD, 91 with simple bronchitis, and 1084 with non-respiratory illnesses (control group)--were studied by collection of five health status indexes and multidimensional assessment. Discriminant analysis was used to identify health status profiles of groups. Demographic, anthropometric, clinical and respiratory function correlates of selected health status profiles were identified. RESULTS: Only 26 and 28% of asthma and COPD patients vs 43% of simple bronchitis and 50% of non-respiratory patients showed group-specific health status profiles. These profiles were characterized by lower 6-min walked distance and greater index of disturbed sleep in asthmatics, and by worse performance on Barthel Index, 6-min walking test and Mini-Mental State Examination in COPD patients. More severe bronchial obstruction, a greater index of comorbidity and a longer occiput-wall distance characterized COPD patients with the worst health status. CONCLUSIONS: The health status of elderly patients with COPD or asthma is highly heterogeneous. On average, COPD is characterized by more severe physical impairment, and asthma by poorer quality of sleep. Comorbidity and severity of bronchial obstruction, but not age, contribute toward defining a subset of COPD patients with the worst health status. PMID- 15132289 TI - Use of a questionnaire to screen for frailty in the elderly: an exploratory study. AB - BACKGROUND AND AIMS: In a pilot study of community-dwelling geriatric clinic patients (N=48, aged 63-90) we examined the use of a questionnaire to classify frailty status by comparing it with standardized markers of frailty. The questionnaire, developed by Strawbridge et al. in 1998, defines frailty as difficulty in more than one of four domains of functioning: physical, cognitive, sensory, and nutritive. METHODS: Subjects were classified as frail or not frail by questionnaire and assignment was compared with testing of physical and cognitive measures in cross-sectional analysis. Demographic variables, functional inventories, physical activity levels, clinician impression of frailty, and 3 year health outcomes were also examined. RESULTS: Thirty-three percent of subjects were classified as frail. Frailty classification by the Strawbridge questionnaire was correlated to Timed Up and Go and repetitive Sit-to-Stand tests, bimanual dexterity and cognitive tests. A discrepancy was found between assignment of cognitive difficulty, by questionnaire and cognitive performance. When overall Strawbridge frailty scores were modified to account for those with poor cognitive performance who did not report cognitive difficulty, the prevalence of frailty increased to 42%. At 3-year follow-up, the modified Strawbridge frailty classification (p<0.05) and clinician impression of frailty (p<0.01) were both significant predictors of death and institutionalization combined. CONCLUSIONS: This study serves as an initial inquiry into the potential validity and utility of the Strawbridge frailty questionnaire as a simple screening tool to identify patients who may warrant detailed functional testing. PMID- 15132290 TI - External hip protectors in home-dwelling older persons. AB - BACKGROUND AND AIMS: External hip protectors have been shown to have a good preventive effect against hip fractures in many studies. However, these studies were carried out either entirely or mostly among older persons living in institutions. The aim of this study was to investigate whether health nurses can, after brief training, choose the right persons to use external hip protectors, so that, with a fairly small number of protectors, the number of hip fractures in home-dwelling older persons in a given area could be significantly reduced. METHODS: One hundred and fifty pairs of external hip protectors were given to the health nurses of the Central district of Tampere Health Center, to give to those older persons aged more than 75 years who, in their opinion, most needed preventive measures against hip fractures. Older persons living in other districts served as controls. The number of hip fractures in each district for each 6-month period was obtained from the Data Center of Pirkanmaa Health District. RESULTS: Whereas in control districts the number of hip fractures increased by 14 from 1.1.-30.6.1998 to 1.1.-30.6.2000 (49-63), their number diminished by 4 (43-39) in the Central district, where the protectors were used. The total cost of the external hip protectors was less than that required to treat one hip fracture in the first year. CONCLUSIONS: External hip protectors are most probably effective and economical in preventing hip fractures also among home-dwelling older persons, as has previously been shown among nursing home patients. PMID- 15132291 TI - Can functional status, after rehabilitation, independently predict long-term mortality of hip-fractured elderly patients? AB - BACKGROUND AND AIMS: Hip fractures are one of the most serious causes of functional impairment and death in the elderly. The aim of this study was to evaluate prospectively the predictive value of functional performance, after rehabilitation, of hip fracture on long-term mortality in community-dwelling patients. METHODS: One hundred and seventy-one patients aged 60 years and over, admitted to a geriatric rehabilitation day unit after inpatient rehabilitation, were followed for up to 4 years. Main outcome measures were Functional Independent Measure (FIM), Timed Get Up and Go test (GUAG), cognitive status using the Mini-Mental State Examination on admission, and mortality during the follow-up period. Kaplan-Meier analysis was carried out on survival curves. RESULTS: All 24 deceased patients performed the GUAG test in > 20 seconds. Although approaching significance, the survival curves were not statistically different between patients performing the test in < or = 20 and those performing it in > 20 seconds (p = 0.08). Survival curves were significantly higher in patients with a FIM score of > or = 90 (p = 0.004), no cardio-cerebrovascular (CCV) diseases (p = 0.001) and no diabetes mellitus (p = 0.01). There were no differences in survival according to age, gender, educational level, marital status, surgical vs conservative treatment, and cognition. A multivariate analysis including FIM score, CCV diseases and diabetes mellitus, demonstrated that only CCV disease was an independent variable for survival (p = 0.02). CONCLUSIONS: Performance, as evidenced by FIM scores after rehabilitation for hip fracture, may provide additional useful information on long-term survival. However, since functional status after rehabilitation is not an independent risk factor for long-term mortality, its predictive value must be interpreted in view of the comorbidities, mainly CCV diseases, which are more important to the risk of mortality than the event of hip fracture itself. PMID- 15132292 TI - Reliability and validity of the Functional Rating Index in older people with low back pain: preliminary report. AB - BACKGROUND AND AIMS: The Functional Rating Index (FRI) was developed to provide an assessment instrument which has not only clinical usefulness but also quantifies the patient's current state of pain and dysfunction in a reliable and valid manner for spinal conditions. There is no study on the FRI applied to older people with low back pain (LBP). The primary aim of this study was to evaluate the validity and reliability of the FRI in older people with LBP. METHODS: A total of 76 subjects aged 65 to 90 years with LBP, of which 37 were cognitively intact and were followed up on a second occasion, were assessed by the FRI, numeric rating scale (NRS), Roland Morris Questionnaire (RMQ) and spinal movement test. Reliability was assessed by statistical analysis of test results for test retest and internal consistency. To assess construct validity, the FRI was compared with the RMQ. Concurrent validity was assessed using the NRS and spinal mobility test. RESULTS: The FRI demonstrated high internal consistency, with alpha=0.921 for test and alpha=0.901 for retest. Item-scale correlations were between 0.549-0.871. Test-retest correlation was 0.913 (p=0.000). There was very good construct validity between the FRI and the RMQ for test (r=0.663, p<0.000) and retest (r=0.603, p<0.000). The FRI showed high correlation with the NRS (r=0.701, p<0.000 for test; r=0.743, p<0.000 for retest) and no correlation with the spinal movement test (r=0.173, p=0.307 for test; r=0.024, p=0.888 for retest). CONCLUSIONS: In this preliminary report, the FRI appears to be easy to administer, seems to have significant validity and reliability, and may be useful in geriatric assessment of older people with LBP. PMID- 15132293 TI - Health benefits associated with exercise habituation in older Japanese men. AB - BACKGROUND AND AIMS: The purpose of this study was to compare the effects of exercise habituation (3-32 years, mean 13.2 years) on physical vitality among five different groups. METHODS: One hundred and two independent, community dwelling elderly Japanese men, aged 64.6 +/- 6.6 years, were recruited as subjects. The vital age test battery consisted of various coronary heart disease risk factors and physical fitness elements. RESULTS: The results of analysis of variance revealed that vital age as an index of physical vitality was youngest in joggers (47.9 yr, N=18), intermediate in trekkers (55.8 yr, N=20) and walkers (59.1 yr, N=18), and oldest (69.6 yr, N=20) in patients with ischemic heart disease (IHD). The difference between chronological age and vital age was approximately 15 years (p<0.05) in joggers, and 8 years (p<0.05) in trekkers and walkers. The vital age of sedentary persons (N=26) was only 1.9 years (NS) younger than their chronological age, which was similar to the difference (vital age of 64.1 +/- 8.5 yr vs chronological age of 65.7 +/- 5.4 yr) previously observed in similarly aged exercising IHD patients. CONCLUSIONS: These results indicate that exercise habituation significantly affects the overall health status of most individuals, irrespective of mode of exercise. Among the three modes of exercise, jogging may be most beneficial. Furthermore, regularly exercising coronary patients may have physical vitality similar to that of sedentary men. PMID- 15132294 TI - Donepezil use in US nursing homes. AB - BACKGROUND AND AIMS: Donepezil is effective in slowing the progression of Alzheimer's disease (AD). No data are available on the patterns of donepezil use among nursing home residents. We estimate the prevalence of the use of donepezil and the frequency of its initiation, and describe the demographic and clinical characteristics of residents taking this drug. METHODS: We used data from the Minimum Data Set (N=174659). We compared users and non-users of donepezil with respect to demographic, clinical and functional variables. We estimated the 6 month incidence of donepezil use. All analyses were stratified by the admission status of the residents (newly admitted/long-stay). RESULTS: Among those meeting the manufacturer's indication for donepezil (mild to moderate AD), the prevalence of use was 30% among newly admitted, and 19% among long-stay residents. About 3% of residents not meeting the manufacturer's indication for donepezil were taking it. The proportion of people with AD still taking donepezil after six months was 44.8% among newly admitted, and 59.5% among long-stay residents. Donepezil use was associated with cognitive impairment, behavioral problems and the use of psychotropic drugs including anti-psychotics and anti-depressants. In people without a diagnosis of AD, there seems to be an association between behavioral problems and use of donepezil. CONCLUSIONS: Donepezil is frequently used in the nursing home setting for the management of AD. Off-label use of donepezil is also relatively frequent. This may reflect an empirical perception of effectiveness on outcomes different from cognitive impairment, such as behavioral problems, although evidence is lacking. PMID- 15132295 TI - The effects of different levels of dietary restriction on non-neoplastic diseases in male Sprague-Dawley rats. AB - BACKGROUND AND AIMS: The primary purpose of the present study was to investigate the effects of 10, 25, and 40% dietary restriction (DR) on non-neoplastic diseases in rodents at 58 and 110 weeks of age, and to determine whether low level DR (10 and 25%) can increase the survival rate and decrease variability in chronic bioassay studies. METHODS: Male Sprague-Dawley (SD) rats (NCTR colony) were divided into four nutritional groups, consisting of an ad libitum (AL) group with unlimited access to the NIH-31 diet, and three dietary restricted (DR) groups given the NIH-31 diet reduced in amount by 10, 25, and 40%. RESULTS: At 110 weeks of age, the incidence of cardiomyopathy was 95, 75, 45, and 15% for AL and 10, 25, and 40% DR rats, respectively; the incidence of nephropathy was 55, 20, 15, and 0% for AL and 10, 25, and 40% DR rats, respectively. The severity of chronic heart and kidney diseases was significantly reduced in all DR rat groups, with significant DR-dependent linear trends for these diseases. Moreover, DR prevented the progression of skin irritation to foot ulcers, and reduced the age related degeneration in the adrenal, lacrimal, and thymus glands, and the liver. CONCLUSIONS: These results clearly indicate that even low DR levels were effective in preventing or slowing the progression of these non-neoplastic diseases. PMID- 15132296 TI - Informed consent for research participation in frail older persons. AB - Informed consent has been the most scrutinized and controversial aspect of clinical research ethics. Institutional review boards (IRBs), government regulatory agencies, and the threat of litigation have all contributed to increasingly detailed consent documents that hope to ensure that subjects are not misled or coerced. Unfortunately, the growing regulatory burden on researchers has not succeeded in protecting subjects, but has rather made the consent process less effective and has discouraged research on vulnerable populations. As a matter of fact, investigators and ethicists continue to identify failures of the consenting process, particularly concerning participation in research of older individuals. The challenges involved in ensuring appropriate consent from the elderly include physical frailty, reduced autonomy and privacy, and impaired decision-making capacity due to dementia, delirium, or other neuropsychiatric illnesses. Ageism among investigators also contributes to failure of informed consent. The evaluation and continuing re-evaluation of an individual's decision making capacity is critical but difficult. In the most extreme cases, the older adult's ability to participate in the consent process is clearly impaired. However, in many instances, the decision-making capacity is only partially impaired but declines during the course of a research project. Implementing methods of effective communication may enable many frail elderly individuals to make informed decisions. Special challenges are posed by research on end-of-life care, which typically involves frail, older subjects who are uniquely vulnerable, and research is conducted in institutional settings where subtle violations of autonomy are routine. Clearly, the frail elderly represent a vulnerable population that deserves special attention when developing and evaluating an informed consent process. Two important ethical conflicts should be kept in mind. First, although vulnerable older patients must be protected, protection should not prevent research on this important population. Similarly, because informed consent documents are often written to prevent legal jeopardy, these technical documents, expressed in language sometimes difficult to understand, can prevent comprehension of basic issues, defeating the ethical purpose of human protection. PMID- 15132297 TI - Prostaglandin I2 does not contribute to the hypotensive effect of the superoxide dismutase mimetic Tempo in rats with aortic coarctation-induced hypertension. AB - This study was designed to investigate the contribution of prostaglandins to the vasodepressor effect of the superoxide dismutase mimetic Tempo in rats made hypertensive by ligation of the abdominal aorta at a point between the left and right renal arteries. Rings of thoracic aorta taken from rats with aortic coarctation released more 6-keto-PGF1alpha (a non-enzymatic product of PGI2 degradation) in the presence than in the absence of Tempo (1 mmol/L; 35.3 +/- 10.1 versus 13.6 +/- 2.6 pg/mg tissue). However, Tempo administered intravenously (2 mg/kg bolus injection plus infusion at 3 mg/kg/h) to rats with aortic coarctation did not increase significantly the concentration of 6-keto-PGF1alpha in vena cava blood. Treatment with Tempo did not affect the arterial pressure of un-operated normotensive rats but promptly decreased the arterial pressure of rats with aortic coarctation-induced hypertension (from 178 +/- 2 to 125 +/- 6 mmHg). The vasodepressor effect of Tempo in hypertensive animals was not affected by pretreatment with indomethacin to inhibit prostaglandin synthesis. These data argue against the hypothesis that PGI2 contributes to the acute hypotensive effect of Tempo in rats with aortic coarctation. PMID- 15132298 TI - Hemodynamic significance of high brachial pulse pressure in young men. AB - This study investigates whether an increased brachial pulse pressure (PP) in young healthy men constitutes a representative measure of the central hemodynamic forces, or the mere expression of an exaggerated upper limb amplification. Thirty two healthy men between 17 and 28 years old underwent noninvasive evaluation of systemic hemodynamics (impedance cardiography) and pulse wave analysis (SphygmoCor). Subjects were divided into 3 predefined groups of brachial PP: < 50, 50-64, and > or = 65 mmHg. The brachial-central PP difference increased with increasing brachial PP (17 +/- 4, 22 +/- 4, and 29 +/- 4 mmHg, respectively P < 0.001). In contrast, peripheral amplification (as measured by the brachial:central PP ratio) remained constant among the tree groups, at the expense of a concomitant widening of aortic PP (P < 0.001) without difference in augmentation index. In the entire sample, central and brachial PP, and the difference between the two measures, correlated positively with cardiac output (P < 0.001) and stroke volume (P < 0.01), and negatively with systemic vascular resistance (P < 0.001), without significant relationship with heart rate. In conclusion, despite the different amplitude, central and brachial PP shared common hemodynamic determinants. A high PP among young men underlied a high output-low resistance circulatory pattern, independently of the site of measurement. PMID- 15132299 TI - Sequence analysis of the fibroblast growth factor 2 gene from the spontaneously hypertensive and hypertrophic heart rats. AB - We have previously reported a quantitative trait locus associated with pressure independent cardiac hypertrophy in the spontaneously hypertensive rat (SHR) of the Okamoto strain. This locus (Lvm1; left ventricular mass locus 1) contains the gene Fgf2 that codes for the potent cardiac growth factor, Fibroblast Growth Factor 2 (FGF2). Given that FGF2 appears essential for the induction of certain forms of cardiac hypertrophy in the rat, we proposed this gene as a candidate for the cardiac enlargement seen in the SHR. Previous reports of elevated FGF2 mRNA levels in the SHR, led us to hypothesise that nucleotide sequence variations occurring in the coding regions or in putative transcriptional factor binding sites within the Fgf2 promoter might play a role in cardiac hypertrophy in this strain. Given that we have also recently derived from the SHR a rat strain that develops spontaneous cardiac hypertrophy in the absence of hypertension (the Hypertrophic Heart Rat; HHR), we also took the opportunity to examine the sequence of its Fgf2 promoter and coding region. However, extensive sequence analysis of the promoter and coding regions of the SHR and HHR Fgf2 genes failed to reveal any nucleotide variations between strains. Thus, we conclude that variations in the nucleotide sequence of the promoter and coding region of the SHR Fgf2 gene do not play a role in the cardiac hypertrophy of the SHR and HHR strains. PMID- 15132300 TI - Free radical activity depends on underlying vasoconstrictors in renal microcirculation. AB - We examined the role of free radicals in renal microvascular tone induced by various vasoactive stimuli. Isolated perfused rat hydronephrotic kidneys were used for direct visualization of renal microcirculation. The effect of tempol on angiotensin II-, norepinephrine-, KCl-, and pressure-induced afferent arteriolar constriction was evaluated. Under angiotensin II-induced constriction, tempol (3 mmol/L) caused 57 +/- 8% dilation of afferent arterioles. In contrast, tempol elicited only 38 +/- 8% and 26 +/- 9% dilation of norepinephrine- and KCl-induced constriction. Similarly, myogenic response induced by elevating renal arterial pressure from 80 to 180 mmHg was resistant to the vasodilator action of tempol (22 +/- 7% inhibition). Furthermore, tempol failed to reverse nitro-L-arginine methylester-induced afferent constriction, nor had vasodilator effect on the angiotensin II-induced constriction in the presence of nitro-L-arginine methylester. In contrast, nitroprusside elicited marked vasodilation of angiotensin II- (97 +/- 5% reversal) and norepinephrine-induced afferent constriction (89 +/- 6% reversal), but had less effect on KCl- (46 +/- 8% reversal) and pressure-induced constriction (26 +/- 9% reversal). These different actions were also observed when polyethylene-glycolated superoxide dismutase was used as an antioxidant. In conclusion, the role of free radicals in afferent arteriolar tone varies, depending on the underlying vasoconstrictor stimuli, with greater contribution of free radicals to angiotensin II-induced constriction. The heterogeneity in the responsiveness to free radical scavengers is attributed to both magnitude of free radicals produced and sensitivity of the underlying vasoconstrictors to nitric oxide. PMID- 15132301 TI - Role of tetrahydrobiopterin in adrenocorticotropic hormone-induced hypertension in the rat. AB - Adrenocorticotropic hormone (ACTH)-induced hypertension in the rat is characterized by nitric oxide deficiency. Tetrahydrobiopterin (BH4) is an essential cofactor for the enzyme nitric oxide synthase and glucocorticoids have been reported to reduce cytokine-induced BH4 production. Accordingly we hypothesized that ACTH-induced hypertension would be reversed by BH4 supplementation. Male Sprague-Dawley rats (n = 33) were treated with BH4 in vehicle (10 mg/kg/day i.p.) or vehicle alone (5 mg/kg/day i.p. of ascorbic acid in 4 mM HCl) for 10 days. ACTH (0.2 mg/kg s.c.) or saline daily injection was started 2 days after BH4 or vehicle treatment and continued for 8 days. Systolic blood pressure (SBP) was measured on alternate days using the tail cuff method. Treatment with HCl, ascorbic acid or BH4 alone had no effect on SBP. In saline treated rats, neither BH4 nor its vehicle modified SBP. In ACTH treated rats, SBP was increased in both BH4 (from 128 +/- 6 to 142 +/- 4 mmHg, T0 to T10, P < 0.0005, one way ANOVA) and vehicle groups (from 127 +/- 3 to 158 +/- 7 mmHg, T0 to T10, P < 0.001, one way ANOVA). There was no significant difference in SBP between BH4 + ACTH treated and vehicle + ACTH treated rats. Thus, daily injection of BH4 (10 mg/kg i.p.) failed to prevent the development of ACTH-induced hypertension in rat. PMID- 15132302 TI - Protein phosphatase 2A B56alpha during development in the spontaneously hypertensive rat. AB - Mistargeting of the regulatory subunit of protein phosphatase 2A (PP2A), B56alpha is involved in the hyperphosphorylation and desensitization of the D1 dopamine receptor in renal proximal tubules of spontaneously hypertensive rats (SHRs). However, the renal expression of B56alpha before hypertension develops is not known. Therefore, we studied the expression of B56alpha and PP2A activity in the kidney during development in the SHR and its normotensive control, the Wistar Kyoto (WKY) rat. PP2A B56alpha was expressed in proximal and distal tubules with no differences in the pattern of expression in WKY and SHRs at any age. In brush border membranes of renal proximal tubules, PP2A B56alpha protein was greatest in the immature rats and decreased with development. However, PP2A activity did not change with age. PP2A B56alpha protein and PP2A activity were similar in WKY and SHRs except at 2 weeks when both PP2A B56alpha protein and PP2A activity were higher in SHRs than in WKY rats. The PP2A catalytic subunit co-immunoprecipitated with the D1 receptor in renal proximal tubule cells. It is possible that the increased expression of PP2A B56alpha and increased basal PP2A activity in the young, especially in the SHRs, may serve as a compensatory mechanism in the increased phosphorylation and decreased renal D1 receptor function, including D1 receptor mediated stimulation in renal proximal tubules of SHRs. PMID- 15132303 TI - The beneficial effect of regular endurance exercise training on blood pressure and quality of life in patients with hypertension. AB - Regular aerobic exercise can reduce blood pressure and is recommended as part of the lifestyle modification to reduce high blood pressure and cardiovascular risk. Hypertension itself, or/and pharmacological treatment for hypertension is associated with adverse effects on some aspects of quality of life. This study was performed to evaluate the effects of regular endurance exercise training on quality of life and blood pressure. Patients with mild to moderate hypertension (systolic blood pressure 140-180 or diastolic blood pressure 90-110 mm Hg) were randomized to a moderate-intensity aerobic exercise group training for 3 sessions/week over 10 weeks or to a non-exercising control group. Health-related quality of life was assessed with the Short Form 36-item Health Survey (SF-36) at baseline and after 6 and 10 weeks. In the 102 subjects (47 male, mean age 47 years) who completed the study, reductions in blood pressure in the exercise group at 10 weeks (-13.1/-6.3 mm Hg) were significant (P < 0.001) compared to baseline and to the control group (-1.5/+6.0 mm Hg). Unlike the control group, the exercise group showed an increase in exercise capacity from 8.2 +/- 1.6 to 10.8 +/- 2.2 METS (P < 0.01) and showed higher scores on 7 out of 8 subscales (P < 0.05) of the SF-36. Improvement in bodily pain and general health sub-scores correlated with reduction in systolic blood pressure. Regular endurance training improves both blood pressure and quality of life in hypertensive patients and should be encouraged more widely. PMID- 15132304 TI - Identification of Nck interacting proteins in vascular smooth muscle cells. AB - The adaptor molecule Nck has been demonstrated to mediate Angiotensin II (AngII) induced stimulation of p21-activated kinase (PAK) and c-Jun NH2-terminal kinase (JNK) in vascular smooth muscle cells (VSMC). We have previously demonstrated, that immunoprecipitation of Nck from VSMC stimulated by AngII yielded an unidentified 100 kD phosphotyrosine (pTyr) protein. The present study was aimed at identifying the Nck-associated 100 kD pTyr protein in VSMC. Several candidate proteins of appropriate size, that had been shown previously either to bind to Nck or had been implicated in signal transduction pathways leading to activation of PAK or JNK were tested for association with Nck in VSMC. The first candidate protein we tested was Git1, which did not bind to Nck in VSMC upon stimulation by AngII. However, we identified dynamin as a 100 kD protein that was bound to Nck in VSCM via interaction with the third Nck-SH3 domain. However, dynamin was not tyrosine phosphorylated by AngII treatment and seemed to be distinct from the 100 kD phosphotyrosine protein that was found in Nck immunoprecipitates. Future work will now have to identify the Nck-associated 100 kD pTyr protein and functional studies will have to address its role in AngII signaling. PMID- 15132305 TI - Accelerometry: providing an integrated, practical method for long-term, ambulatory monitoring of human movement. AB - Accelerometry offers a practical and low cost method of objectively monitoring human movements, and has particular applicability to the monitoring of free living subjects. Accelerometers have been used to monitor a range of different movements, including gait, sit-to-stand transfers, postural sway and falls. They have also been used to measure physical activity levels and to identify and classify movements performed by subjects. This paper reviews the use of accelerometer-based systems in each of these areas. The scope and applicability of such systems in unsupervised monitoring of human movement are considered. The different systems and monitoring techniques can be integrated to provide a more comprehensive system that is suitable for measuring a range of different parameters in an unsupervised monitoring context with free-living subjects. An integrated approach is described in which a single, waist-mounted accelerometry system is used to monitor a range of different parameters of human movement in an unsupervised setting. PMID- 15132306 TI - Pseudo-hypertension and arterial stiffness: a review. AB - Hypertension is a condition of persistently elevated blood pressure, associated with increased cardiovascular risk. Non-invasive BP measurement using Korotkoff sounds is the most common method of screening for the condition. The possibility of inaccurate readings leading to a false diagnosis of hypertension (pseudo hypertension) is of concern. Stiffened arteries in the elderly have been proposed as being the primary cause of pseudo-hypertension. Non-invasive detection of pseudo-hypertension remains problematic. This paper reviews clinical literature on pseudo hypertension and approaches to measuring the compressive stiffness of arteries, as well as biomechanical literature regarding models of arterial stiffness and the origin of Korotkoff sounds. Models of the latter show the importance of the relationship between transmural pressure and cross-sectional area (P1/Csa curve) of the brachial artery as it closes under the influence of the pressure cuff. The review concludes that future research on pseudo hypertension should include development of new instrumentation to measure the P1/Csa curve of the brachial artery in vivo using non-invasive techniques suitable for application to an elderly population. PMID- 15132307 TI - Effect of cooling and re-warming on cerebral and whole body electrical impedance. AB - Cerebral electrical impedance is useful for the detection of cerebral edema following hypoxia in newborn infants. Thus it may be useful for determining neurological outcome or monitoring treatment. Hypothermia is a promising new therapy currently undergoing trials, but will alter impedance measurements. This study aimed to define the relationship between temperature and both cerebral and whole body electrical impedance, and to derive correction factors for adjustment of impedance measurements during hypothermia. In eight anaesthetized 1-2 day old piglets rectal, tympanic and scalp temperatures were monitored continuously. Following baseline readings at a rectal temperature of 39 degrees C, piglets were cooled to 32 degrees C. Four piglets were re-warmed. Cerebral and whole body impedance were measured at each 0.5 degrees C as rectal temperature decreased. There was a strong linear relationship between both cerebral and whole body impedance and each of the temperatures measured. There was no difference in the relationship between impedance and rectal, tympanic or scalp temperatures. The relationship for impedance and rectal temperature was the same during cooling and re-warming. Using the correction factors derived it will be possible to accurately monitor cerebral and whole body fluid distribution during hypothermic treatment. PMID- 15132308 TI - A new biomedical sensor for measuring PCO2. AB - Measuring PCO2 (partial pressure of carbon dioxide) in an organ can enable early detection of ischemia. However, there are few clinical applicable solutions for measuring PCO2. Based upon the requirement for clinical applications, a conductivity based PCO2 sensor is proposed. A conductivity based PCO2 sensor measures conductance in an aqueous solution separated from the measured object by a gas-permeable membrane. A bridge design with two cavities is favored for such a sensor. A planar and a cylindrical macro prototype based upon the bridge design were studied. The design criteria were based on the contribution from the electrode polarization, stray capacitances, contact area with the sample and design ability to miniaturize the sensor. The cylindrical sensor is favored because of its large contact area and advantages for miniaturization. Further investigation has to be done to confirm the functionality of such a design in a miniaturized form and its clinical performance. PMID- 15132309 TI - Investigation of cerebral haemodynamics by near-infrared spectroscopy in young healthy volunteers reveals posture-dependent spontaneous oscillations. AB - Autonomic reflexes enable the cardiovascular system to respond to gravitational displacement of blood during changes in posture. Spontaneous oscillations present in the cerebral and systemic circulation of healthy subjects have demonstrated a regulatory role. This study assessed the dynamic responses of the cerebral and systemic circulation upon standing up and the posture dependence of spontaneous oscillations. In ten young healthy volunteers, blood pressure and cerebral haemodynamics were continuously monitored non-invasively using the Portapres and near-infrared spectroscopy (NIRS), respectively. Oscillatory changes in the cerebral NIRS signals and the diastolic blood pressure (DBP) signal have been identified by the fast Fourier analysis. Blood pressure increased during standing and returned to basal level when volunteers sat on a chair. The mean value of cerebral tissue oxygen index (TOI) as measured by NIRS did not demonstrate any significant changes. Oscillatory changes in DBP, oxyhaemoglobin concentration [O2Hb] and TOI showed a significant increase when subjects were standing. Investigation of the low frequency component (approximately 0.1 Hz) of these fluctuations revealed posture dependence associated with activation of autonomic reflexes. Systemic and cerebral changes appeared to preserve adequate blood flow and cerebral perfusion during standing in healthy volunteers. Oscillatory changes in [O2Hb] and TOI, which may be related to the degree of cerebral sympathetic stimulation, are posture dependent in healthy subjects. PMID- 15132310 TI - Validation of a new dielectric device to assess changes of tissue water in skin and subcutaneous fat. AB - Easily applicable and inexpensive water-specific techniques to evaluate local oedema, swollen tissue problems and fluid retention in humans are not available. In the present investigation a recently constructed non-invasive device for a local measurement of changes in tissue water in human skin and subcutaneous fat (SSF) was validated. The instrument transmits an ultra high-frequency electromagnetic (EM) wave of 300 MHz into a coaxial line and further into an open ended coaxial probe which is in contact with the skin. Due to the dimensions of the applied probe the penetration of the EM field extends to subcutaneous fat. A major part of the EM energy is absorbed by tissue water while the rest is reflected back into a coaxial line. From the information of the reflected wave an electrical parameter, directly proportional to tissue water content, called a dielectric constant of SSF, was calculated. For system validation, the decrease of water content in SSF measured with the dielectric technique in the volar forearm of seven patients during haemodialysis treatment was compared with the decrease of the circumference of the forearm and the amount of fluid removed. Statistically highly significant correlations were obtained between the decreasing dielectric constant (i.e. water content) of the SSF and the fluid removed during haemodialysis treatment (r = -0.99, p < 0.01) and between the decreasing dielectric constant and the circumference of the arm (r = 0.97, p < 0.05). The sensitivity of the dielectric method was four-fold compared with the circumferential measurement. The repeatability 3.0% was not dependent on the phase of haemodialysis. The new device allows an easy and non-invasive measurement technique to assess changes of tissue water in SSF. PMID- 15132311 TI - In vitro assessment of a continuous cardiac output catheter system. AB - Continuous measurement of cardiac output (CCO) is useful in assessing the cardiovascular status of patients during cardiac surgery and in intensive care. Recently, a CCO system (truCCOMS, Aortech, UK), capable of detecting rapid changes in cardiac output (CO) was introduced. The method is based on the energy required to maintain an integral heat-transfer device at constant temperature above the ambient value. The aim of this study was to assess the performance of this CCO system in vitro under in steady as well as pulsatile flow conditions representative of those in the pulmonary artery. In order to determine the sensitivity of the system to changes in vessel cross-sectional area and therefore local flow velocity, the catheter was deployed in a linear-tapered tube. Steady and pulsatile flows were generated, and the electrical power at various locations along the tapered tube was recorded. The results show significant differences in the performance under the two different flow conditions. In steady flow, the CO was highly dependent on the local velocity whereas in pulsatile flow, CO varied much less with local velocity. The sensitivity expressed as a percentage increase in CO per 100% increase in velocity at a CO of 5 l min(-1) was 87% in steady flow and 24% in pulsatile flow. Experiments carried out with three fluids with different viscosity show that the errors in determining CO in the tapered tube were also dependent on the Reynolds number and flow regime. The mean errors ranged from about 50% at 2 l min(-1) to less than 10% at 8 l min(-1). The correlation between the predicted and actual CO was generally good. In conclusion, the pulmonary artery catheter is not recommended in situations where blood flow is expected to be steady or of low pulsatility. It may, however, be suitable under normal pulsatile flow conditions in the pulmonary artery. PMID- 15132312 TI - Comparison of caffeine-induced changes in cerebral blood flow and middle cerebral artery blood velocity shows that caffeine reduces middle cerebral artery diameter. AB - Changes in cerebral blood flow (CBF) can be assessed directly with xenon clearance (XeC) or indirectly by measuring changes in middle cerebral artery blood velocity (Vmca) with transcranial Doppler (TCD). The aim of this study was to compare the changes in CBF and Vmca following caffeine ingestion. Nineteen patients (age 48-86, recovering from an acute stroke) and ten controls (age 52 85) were each studied twice. Bilateral measurements of CBF and Vmca were made before and after ingestion of 250 mg caffeine or matched placebo. The percentage change in CBF and Vmca after caffeine was calculated. Full results (CBF and Vmca) were obtained from 14 patients and 9 controls. There was no significant difference between patients and controls, so results were combined. Caffeine reduced CBF by 22% (95% confidence interval (CI) = 17% to 28%) and reduced Vmca by 13% (95% CI = 10% to 17%). The fall in Vmca was significantly less than that in CBF (p = 0.0016), showing that caffeine reduces mca diameter. Analysis based on Poiseuille flow in the arterioles suggests that caffeine reduced arteriole diameter by 5.9% (95% CI = 4.6% to 7.3%) and mca diameter by 4.3% (95% CI = 2.0% to 6.6%). TCD is being used as an alternative to XeC for assessing the effect of vasoconstrictors and vasodilators on CBF. This study has demonstrated that in mca diameter can be changed by the vasoactive agents, and that changes in Vmca do not necessarily reflect changes in CBF. PMID- 15132313 TI - Improved lung edema monitoring with coronary vein pacing leads: a simulation study. AB - This computer simulation study compared the ability of left ventricular coronary vein (LV) pacemaker leads against right ventricular (RV) and right atrial (RA) leads to monitor lung edema using electrical impedance measurements. MRI images were used to construct electrical models of the thorax. Four lead configurations were tested with increases of pulmonary edema, intravascular fluids and heart dilation. The impedance changes observed at end systole with severe lung edema were 8.5%, 11.2%, 12.3% and 26.8% for the RA, RV, RV coil and LV configurations, respectively. Sensitivities in ohms per litre of lung fluid were 19.15, 19.15, 25.07 and 52.11 for the same configurations. The impedance changes for intravascular fluid overload with constant lung status were 1%, 1.3%, 9.2% and 6.4% while the sensitivities were 2, 2, 17 and 11 ohms per litre of intravascular fluid, respectively. Regional analysis of the thoracic sources of impedance revealed a high sensitivity near pacing electrodes and generator, and a low sensitivity to the right lung and all pulmonary vessels. Simulations showed that LV leads have a threefold advantage in sensitivity when monitoring lung edema in comparison to conventional RV leads. To monitor vascular and lung fluids independently, combined impedance configurations may be used. Regional sensitivities must be taken into account for proper clinical interpretation of impedance changes. PMID- 15132314 TI - Bias and uncertainty in heart rate variability spectral indices due to the finite ECG sampling frequency. AB - Spectral analysis of the heart rate variability is becoming a usual tool as a marker of the autonomic nervous system. The final output of the spectral analysis is a set of indices that are always estimators due to technical limitations. In this work, the bias and the uncertainty in the VLF, LF, HF and LF/HF indices due to the finite sampling frequency of the ECG are analysed. The results indicate that for low sampling frequency (125 Hz), the bias and uncertainty in the HF and LF/HF indices can blur the results of the analysis, especially if the RR time series has low variability. The HF index is overestimated and, accordingly, the LF/HF index is underestimated. Then, results from RR time series with low sampling frequency must be used with care. The uncertainty of the spectral indices is proportional to the inverse of the sampling frequency and the bias is proportional to the inverse of the square sampling frequency. PMID- 15132315 TI - Change in the static rheological properties of the aorta in Kurosawa and Kusanagi hypercholesterolemic (KHC) rabbits with progress of atherosclerosis. AB - The rheological properties of the arterial wall have intimate connections with the fine structure of the wall. Alteration in fine structure due to cardiovascular disease, such as atherosclerosis, could affect the rheological characteristics of the wall. The present study was designed to investigate changes in the static rheological properties of the aorta in Kurosawa and Kusanagi-Hypercholesterolemic (KHC) rabbits aged 10-12, 22-24 and 34-36 months in relation to histological alteration of the wall due to progression of atherosclerosis with age. Circumferential wall strips were excised from the ascending, proximal descending thoracic and proximal abdominal aortas and their stress/strain relationship was recorded. Tensile force of the wall showed a slight but insignificant decrease in the KHC rabbit group aged 10-12 months compared to that in the age-matched control group in the proximal thoracic aorta and increased significantly with ageing in the KHC rabbits in these aortic regions mainly at medium and high strain ranges. Wall stress was significantly smaller in the 10-12 months old KHC rabbit group than in the age-matched control group in the proximal thoracic and proximal abdominal aortas and increased significantly with ageing in the KHC rabbit groups chiefly at medium and high strain ranges. Incremental elastic modulus determined at 50% stretching of the initial length of the wall strip was also significantly lower in the KHC rabbit group aged 10-12 months in comparison to that in the age-matched control group and increased significantly with ageing in the KHC rabbit group. The intima thickened severely with abundant foam cells in the KHC rabbits aged 10-12 months. With increasing age, collagen and elastin fibres showed signs of gradual proliferation among the foam cells. The aortic wall in KHC rabbits was viscoelastic in the relatively early stage of atherosclerosis due to abundant foam cells, and thereafter increased in stiffness gradually with fibrous proliferation and calcification. We can conclude that the static rheological properties of the atherosclerotic aortic wall changed in association with alteration in the microstructure of the wall with progression of atherosclerosis. PMID- 15132316 TI - Effects on baroreflex sensitivity measurements when different protocols are used to induce regular changes in beat-to-beat intervals and systolic pressure. AB - Baroreflex sensitivity is becoming an important clinical measurement. Nevertheless there is no recommend standard measurement protocol. This study assessed the ability of eight protocols to induce regular changes in cardiac beat to-beat interval and systolic pressure (SP), and the effect each protocol had on baroreflex sensitivity (BRS). Twelve subjects had changes in cardiac beat-to-beat intervals and SP levels induced at 8 times a minute by following 8 different protocols, each for 3 min. These comprised breathing in a supine and standing posture, breathing through a resistance, breathing into a closed orifice (the breathing protocols), and performing handgrip exercises, being rocked, having legs raised and lowered, and being presented with mental arithmetic questions (the non-breathing protocols). Induction success of each protocol was determined by the percentage of cardiac beat-to-beat interval and SP level signals with a peak at 8 times per minute in their frequency spectra. The consistency of the induced changes was measured by a signal-to-noise ratio (SNR). BRS was calculated from the frequency spectra. The induction success was 85% for breathing and 31% for non-breathing protocols. The consistency of cardiac beat-to-beat interval changes was highest with supine breathing (SNR = 1.6 +/- 0.3) and resistance breathing (SNR = 1.5 +/- 0.5) protocols. The consistency of SP level changes was highest with resistance breathing (SNR = 1.0 +/- 0.3) and breathing into a closed orifice (SNR = 1.0 +/- 0.5) protocols. BRS values in the supine breathing protocol (24 +/- 10 ms mmHg(-1)) and the handgrip protocol (32 +/- 3 ms mmHg(-1)) were significantly greater (p < 0.05) than for standing breathing (11 +/- 5 ms mmHg(-1)), resistance breathing (17 +/- 8 ms mmHg(-1)) or breathing into a closed orifice (12 +/- 5 ms mmHg(-1)) protocols. Different protocols have different induction successes and degrees of effectiveness in inducing cardiac beat-to-beat and SP level changes. BRS is affected by the induction protocol used, highlighting the need for a standard measurement protocol. PMID- 15132317 TI - Reproducibility and reliability of fetal cardiac time intervals using magnetocardiography. AB - We investigated several factors which may affect the accuracy of fetal cardiac time intervals (CTI) determined in magnetocardiographic (MCG) recordings: observer differences, the number of available recording sites and the type of sensor used in acquisition. In 253 fetal MCG recordings, acquired using different biomagnetometer devices between the 15th and 42nd weeks of gestation, P-wave, QRS complex and T-wave onsets and ends were identified in signal averaged data sets independently by different observers. Using a defined procedure for setting signal events, interobserver reliability was high. Increasing the number of registration sites led to more accurate identification of the events. The differences in wave morphology between magnetometer and gradiometer configurations led to deviations in timing whereas the differences between low and high temperature devices seemed to be primarily due to noise. Signal-to-noise ratio played an important overall role in the accurate determination of CTI and changes in signal amplitude associated with fetal maturation may largely explain the effects of gestational age on reproducibility. As fetal CTI may be of value in the identification of pathologies such as intrauterine growth retardation or fetal cardiac hypertrophy, their reliable estimation will be enhanced by strategies which take these factors into account. PMID- 15132318 TI - Compression of the brachial artery in vivo. AB - Stiffening of the brachial artery is implicated in pseudo-hypertension. To date, a reliable clinical predictor of the condition has not been developed. This paper describes the development of prototype instrumentation and methodology for measurement of the brachial artery transmural pressure/cross-sectional area relationship in vivo. The methodology has been validated using a model of an arm and a thin-walled rubber tube. Application of the technique to a healthy subject shows that the technique is viable and gives good reproducibility. Closure of the brachial artery with reducing transmural pressure is observed and recorded. The new technique has some important advantages over existing methodologies. PMID- 15132319 TI - Assessment of parallel conductance for the trans-cardiac conductance method: can we use the hypertonic saline method with pulmonary artery injections? AB - The trans-cardiac conductance (TCC) method provides on-line left ventricular (LV) volume signals by determining the electrical conductance of blood in the LV using central venous and epithoracic electrodes. Conductive structures outside the LV cause a 'parallel conductance' offset term (Vp) that is determined by bolus injections of hypertonic saline in the pulmonary artery (Vp(saline)). Analysis of the increased conductance signal during passage of the bolus through the LV yields Vp(saline). Since TCC signals are picked up by epithoracic electrodes, concern has been raised that hypertonic saline remaining in the lungs might lead to overestimation. The decrease in blood conductivity induced by injection of non ionic contrast medium during a LV angiogram may also be used to determine Vp (Vp(contrast)). Since the contrast is injected directly into the LV, lung conductance should be unaltered. Thus, we compared Vp(saline) with Vp(contrast) in six anaesthetized sheep during different hemodynamic conditions. Linear regression showed that Vp(saline) = 0.99 Vp(contrast) + 2.45 ml (r2 = 0.99). Bland-Altman analysis yielded a small non-significant bias (+/-2SD) of 1.8 (+/ 6.8) ml. We conclude that parallel conductance for TCC can be accurately determined with the conventional hypertonic saline method. PMID- 15132320 TI - Monitoring nasal conductance by bilateral nasal cannula pressure transducers. AB - Nasal obstruction is common and may disturb well-being during daytime and sleep. Its objective assessment by standard methods requires patient cooperation and inconvenient instrumentation. Therefore, the purpose of this study was to develop a novel, unobtrusive technique for continuous monitoring of changes in nasal conductance during natural breathing based on pressure recordings at the left and right nares. In ten volunteers, pressure swings derived independently from left and right nasal cannula were recorded at baseline, and after topical application of histamine and xylomethazoline. Nasal conductance was measured by means of epipharyngeal pressure recordings and face-mask pneumotachography as the reference standard. Decreases in nasal conductance were associated with changes in relative timing and shape of left and right nasal cannula derived pressure swings during breathing. This was reflected in decreases in the coefficient of cross-correlation between left and right nasal pressure signals. Diagnostic accuracy in detecting a fall in nasal conductance to <80% baseline by the maximal coefficient of determination from cross-correlation of left versus right nasal pressure was evaluated by receiver operator characteristics. The area under the curve was 0.88, 95% confidence interval 0.76 to 0.99, n = 40, P < 0.05 versus 0.5. We conclude that patterns of pressure swings derived from left and right nasal cannula may serve as a noninvasive means to detect changes in nasal conductance without requirement of patient cooperation. PMID- 15132321 TI - Beat-to-beat detection of fetal heart rate: Doppler ultrasound cardiotocography compared to direct ECG cardiotocography in time and frequency domain. AB - In order to obtain power spectral information on the fetal heart rate in stages of pregnancy earlier than labor an algorithm has been developed to calculate the fetal heart rate on a beat-to-beat basis from Doppler ultrasound cardiotocographic signals. The algorithm was evaluated by comparing the calculated fetal heart rate with the heart rate determined from direct ECG signals measured with a scalp electrode. Heart rates were compared both in time and frequency domain. In the time domain the results achieved by both methods correlate well (correlation coefficient = 0.977 (p < 0.001)), in the frequency domain the results correlate even better (correlation coefficient = 0.991 (p < 0.001)). Based on these findings, it can be concluded that the developed algorithm provides a valuable tool for obtaining power spectral information on the fetal heart rate in stages of pregnancy earlier than labor. PMID- 15132322 TI - A novel mounting device for attaching intracranial probes to the skull for use in experimental research models. AB - A simple mount capable of securely holding a variety of intracranial probes to the skull was constructed from commonly available clinical consumables. Using this device the cerebral cortical blood flow of preterm lambs was measured using a laser Doppler flow probe, and cerebral pH and cortical electrical impedance were measured in newborn piglets using pH electrodes and Ag/AgCl wire electrodes. In both studies, the mount held the various probes for periods up to 6 h with no dislodgement or probe failure. The simple mount presented here can be adapted to a wide variety of intracranial probes and will hold them securely to the skull. PMID- 15132323 TI - Total ankle arthroplasty. PMID- 15132324 TI - Theory and technique of translational manipulation for adhesive capsulitis. AB - Manipulation of the glenohumeral joint with the patient under anesthesia has long been used as a treatment modality for refractory shoulder stiffness. Recently, translational manipulation of the glenohumeral joint under interscalene brachial plexus regional block has been shown to be efficacious in the treatment of adhesive capsulitis and to present a low risk of iatrogenic injury. The theory and technique of glenohumeral translational manipulation for the treatment of adhesive capsulitis are presented here. PMID- 15132325 TI - Current recommendations for extended out-of-hospital thromboprophylaxis following total hip arthroplasty. AB - Patients who have undergone total hip arthroplasty are at risk for venous thromboembolic disease, including deep venous thrombosis, pulmonary embolism, and postphlebitic syndrome. Although the number of annual total hip arthroplasties in the United States has steadily risen in the past decade, the rate of venous thromboembolic disease within that patient group has declined as a result of advances in surgical and postoperative procedures, including prompt mobilization and safety and efficacy of thromboprophylactic alternatives. This paper reviews the therapeutic options for deep venous thrombosis prophylaxis following total hip arthroplasty, with an emphasis on recommendations of the Sixth American College of Chest Physicians Consensus Conference on Antithrombotic Therapy. The implications of recent randomized clinical trials on care provided by physiatrists in the rehabilitation facility setting are also discussed. The low molecular-weight heparins have become widely accepted alternatives to aspirin, heparin, and warfarin as results from randomized clinical trials have proved their superior safety and efficacy in the postoperative period. Prolonged prophylaxis up to 30 to 35 days postoperatively in total hip arthroplasty may offer additional protection against venous thrombosis. Therefore, to prescribe optimal thromboprophylactic regimens following total hip arthroplasty, physicians require an understanding of the current recommendations of the Sixth American College of Chest Physicians Consensus Conference on Antithrombotic Therapy and the implications of recently concluded clinical trials. PMID- 15132327 TI - Primary exchange revision arthroplasty for infected total knee replacement: a long-term study. AB - Infected knee replacement is a serious complication that requires significant hospital-based resources for successful management. A successful primary exchange revision technique offers decreased morbidity for the patient by eliminating a second major operation and associated hospitalization, which in turn substantially reduces the cost associated with this dreaded condition. Twenty-two consecutive infected primary total knee arthroplasties in 22 patients were treated with primary exchange revision arthroplasties using antibiotic impregnated cement. The surgical technique consisted of excision of draining sinuses, complete synovectomy, removal of granulation tissue, and debulking of the extensor mechanism. Postoperative treatment consisted of 4 to 6 weeks of parenterally administered antibiotics and 6 to 12 months of orally administered antibiotics. Both gram-positive and gram-negative organisms were treated. At an average follow-up of 10.2 years (range, 1.4 to 19.6 years), 90.9% were free of recurrent infection. Knee scores averaged 79.5, with 85.7% good or excellent results. The physiological classification of the host appeared to influence the outcome of revision. All patients in class A or B (20 knees) had successful eradication of their primary infection after 1-stage revision, whereas the only failure in the group was a physiological class C patient, who eventually expired from ongoing end-organ liver failure. These results compare most favorably with those of delayed-exchange revision arthroplasty while providing a more cost effective management program. PMID- 15132326 TI - Bilateral cobalt alloy femoral component fracture: a case report. AB - Even though at present femoral component fracture is a rare complication of total hip replacement, conditions still exist that predispose the prosthesis to failure. Component failure should be considered when a patient presents with pain in a previously asymptomatic hip, particularly in complex dysplastic hips requiring small stems, trochanteric osteotomy, and compromised cement technique. PMID- 15132328 TI - Traumatic superficial temporal artery pseudoaneurysms in a minor league baseball player: a case report and review of the literature. AB - Traumatic STA aneurysm is a rare complication of facial trauma occuring typically in young men. We present the case of a minor league baseball player who developed 2 pseudoaneurysms after being struck by a baseball and review all cases associated with sports activities. Reports associated with sports activities are increasing and may represent an increasing incidence. The team physician should suspect this condition when a player presents with a new temporal mass after facial trauma. Diagnosis is typically made on history and physical examination, but can be confirmed by duplex ultrasound. Definitive treatment is surgical resection of the aneurysm after proximal and distal ligation of the vessel. PMID- 15132329 TI - Diaphyseal aclasis. PMID- 15132330 TI - A simplified method for sampling and analysis of high volume surface water for organic contaminants using XAD-2. AB - A simple compressed-gas driven system for field processing and extracting water for subsequent analyses of hydrophobic organic compounds is presented. The pumping device is a pneumatically driven pump and filtration system that can easily clarify at 4 L/min. The extraction device uses compressed gas to drive filtered water through two parallel XAD-2 resin columns, at about 200 mL/min. No batteries or inverters are required for water collection or processing. Solvent extractions were performed directly in the XAD-2 glass columns. Final extracts are cleaned-up on Florisil cartridges without fractionation and contaminants analyzed by GC-MS. Method detection limits (MDLs) and recoveries for dissolved organic contaminants, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and pesticides are reported along with results of surface water analysis for the San Francisco Bay, CA. PMID- 15132331 TI - Analysis of organochlorine pesticides in water by novel activated carbon fiber solid phase microextraction coupled with gas chromatography-mass spectrometry. AB - This study describes a fast activated carbon fiber-solid phase microextraction (ACF-SPME) method for determining organochlorine pesticides (OCPs) in water. The pesticides in this study consist of Hexachlorobenzene (HCB) and alpha-, beta-, gamma-hexachlorocyclohexanes (HCHs). The optimal experimental procedures for the adsorption and desorption of four OCPs were evaluated. The linearity was obtained with a RSD of 20% for the OCPs studied over a range from 1.0 to 100 microg/L. The limits of detection at ng/L level were achieved with GC-MS under selected ion monitoring (SIM) acquisition mode. The proposed method was applied to the determination of OCPs concentration in tap water. The results have demonstrated the suitability of the ACF-SPME-GC-MS approach for the analysis of multi-residue OCPs in water. Compared to the commercial fiber, ACF has shown its advantages in solvent-resistance, thermal stability, and the cost. The results obtained in this study suggest that ACF is a promising choice in solid phase microextraction. PMID- 15132332 TI - Analysis of herbicide Krovar I by liquid chromatography with atmospheric pressure chemical ionization mass spectrometry. AB - A simple, very efficient method is presented for routine analysis of herbicide Krovar I (active components bromacil and diuron) in water and soil samples. Water samples were extracted by liquid-liquid extraction with dichloromethane (DCM) as extraction solvent. For soil samples two different extraction techniques were compared: microwave-assisted solvent extraction and a shaking technique using a platform shaker. Extracts were analyzed by high performance liquid chromatography using a water:methanol gradient. Liquid chromatography was coupled with atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS) for quantification of bromacil and diuron. Optimization of the APCI-MS was done by using standards in the flow injection analysis mode (FIA). Method detection limit for liquid samples for bromacil is 0.04 microg L(-1) and for diuron 0.03 microg L(-1). Method detection limit for soil samples is 0.01 microg g(-1) dry weight for both compounds. Results of analysis of field samples of water and soil are also presented. PMID- 15132333 TI - Prediction of PCB content in sportfish using semipermeable membrane devices (SPMDs). AB - Triolein-filled semipermeable membrane devices (SPMDs) were immersed at three locations along the St. Joseph River in northern Indiana for 30 days to see if the PCB content of fish from the same location could be predicted with this model device. Triolein from the SPMD's was analyzed for PCB using enzyme-linked immunosorbent assay (ELISA) and compared to residues detected in fish collected from the same locations. There was a significant difference (p < 0.05) in total PCB concentrations between SPMD samples. However, due to variability in PCB residues between species and low PCB residues in SPMDs, a direct correlation between PCBs in sportfish and SPMDs could not be determined. PMID- 15132334 TI - DDT residues in Lebanese soils. AB - The present study was conducted to evaluate the levels of residual DDT in Lebanese soils. A total of 113 surface soil samples were collected for analysis from three major agricultural regions in the country: Mount Lebanon, Beqa'a valley and the coastal plain. The values of residual DDT in soils ranged between 0 and 1190 ng g(-1). The majority of the analyzed soil samples were free or contained very low concentrations of DDE. Only two surface soil samples, which were collected from urban areas with high human activities, contained relatively high concentrations of residual DDT. The levels of residual DDT in all of the analyzed soil samples were within the permissible limits for agricultural soils. PMID- 15132335 TI - Photocatalytic degradation of an organophosphorus pesticide phosalone in aqueous suspensions of titanium dioxide. AB - The present work deals with photocatalytic degradation of an organophosphorus pesticide, phosalone, in water in the presence of TiO2 particles under UV light illumination (1000 W). The influence of the basic photocatalytic parameters such as pH of the solution, amount of TiO2, irradiation time, stirring rate, and distance from UV source, on the photodegradation efficiency of phosalone was investigated. The degradation rate of phosalone was not high when the photolysis was carried out in the absence of TiO2 and it was negligible in the absence of UV light. The half-life (DT50) of a 20 ppm aqueous solution of phosalone was 15 min in optimized conditions. The plot of lnC (phosalone) vs. time was linear, suggesting first order reaction (K=0.0532 min(-1)). The half-life time of photomineralization in the concentration range of 7.5-20 ppm was 13.02 min. The efficiency of the method was also determined by measuring the reduction of Chemical Oxygen Demand (COD). During the mineralization under optimized conditions, COD decreased by more than 45% at irradiation time of 15 min. The photodegradation of phosalone was enhanced by addition of proper amount of hydrogen peroxide (150 ppm). PMID- 15132337 TI - Ecological risk assessment of neem-based pesticides. AB - A tiered process was used to evaluate the risks of pure azadirachtin (AZA) and two neem-based insecticides (Neemix and Bioneem) on six aquatic animals [crayfish (Procambarus clarkii), white shrimp (Penaeus setiferus), grass shrimp (Palaemonetes pugio), blue crabs (Callinectes sapidus), water fleas (Daphnia pulex), and mosquito larvae (Culex quinquefasciatus)] through short term acute toxicity tests. The risk was calculated using the level of concern endpoints (Q values) and relative hazard index (RHI) for acute and chronic exposure scenarios. The Q values of Neemix, Bioneem, and pure AZA derived from acute exposure tests indicated that D. pulex is the only sensitive species to the test pesticides. Furthermore, the RHI values of Neemix and Bioneem for D. pulex were above the critical limit of 10 indicating that these pesticides may pose a moderate hazard to this species and related crustaceans in acute exposure scenarios. The RHI values of the two pesticides and pure AZA were all below the critical limit of 10 for P. clarkii, P. setiferus, P. pugio, C. sapidus, and C. quinquefasciatus. The aquatic risk assessment process showed that the risk values of tested pesticides did not exceed the criteria, and therefore, no ecological hazard is likely to result from their use. PMID- 15132336 TI - A study on the environmental degradation of pesticides azinphos methyl and parathion methyl. AB - The effect of environmental parameters (temperature and relative humidity) on the degradation rate of azinphos methyl and parathion methyl was studied. Proprietary emulsifiable concentrates were diluted and added to each of 90 glass Petri dishes for each pesticide and were left overnight to dry. Petri dishes were placed in 18 air-tight containers (9 for each pesticide) in which were created environments with relative humidity (RH) of 60, 82, and 96%. The containers were stored at 0, 20, and 40 degrees C. From the experimental results best fit curves, kinetic equations, rate constants, and half-lives were calculated. Half-lives of azinphos methyl for the RH studied were, from 124 to 267 days at 0 degrees C, from 89 to 231 days at 20 degrees C, and from 25 to 71 days at 40 degrees C. Corresponding half-lives for parathion methyl were from 48 to 57 days at 0 degrees C, from 9.2 to 10.5 days at 20 degrees C and from 1.3 to 1.5 days at 40 degrees C. The results were correlated with relevant results from the decomposition of the same or similar pesticides on apples both, on the trees and during refrigerated storage. These correlations are suggesting that biological factors strongly affected the decomposition rate of azinphos methyl. On the contrary the decomposition of parathion methyl was mainly affected by environmental rather than biological factors. PMID- 15132338 TI - Bioaccumulation kinetics and bioconcentration factor of chlorinated pesticides in tissues of Puntius ticto (Ham.). AB - Bioaccumulation kinetics and bioconcentration factor (BCF) of chlorinated pesticides like Aldrin, Dieldrin, Benzene hexachloride (BHC), and Dichlorodiphenyl-dichloro-ethane (DDT) in fish tissues of Puntius ticto was studied in detail in a continuous fed system. The bioconcentration process is summarized by using a first order uptake model and the steady-state BCF is calculated based on the 30 days exposure. Rate of bioaccumulation of DDT was maximum of 4.6432 microg g(-1) wet weight per day in liver tissue whereas it was minimum of 0.0002 microg g(-1) wet weight per day in case of Dieldrin in the muscle tissue among the pesticides. It was observed that DDT showed maximum BCF of 89.010 in case of liver tissue of the fish exposed to 30 days. The regression coefficient (r2) between pesticide concentration and exposure time varied between 0.6212 and 0.9817 indicating high correlation. Based on actual calculated BCF values, the octanol water partition coefficient (Kow) values were predicted. In order to prove the hydrophobic property of chlorinated compounds and its affinity towards lipid, the Kow is predicted. Results showed that pesticide burden differ from tissue to tissue and can be correlated to the lipid content, size, exposure time, and species. PMID- 15132339 TI - Are physicians and medical students prepared to educate patients about alcohol consumption? AB - Given the tremendous financial and human costs of alcohol abuse and dependence, physician competency in alcohol related issues must be increased. Alcohol competency is essential to evaluate the risks and to detect alcohol problems early in the course of alcohol abuse and dependence. But, are medical students and physicians adequately trained in alcohol-related issue to give appropriate advice about alcohol? Are they trained to use an office visit to promote health, identify early abuse and dependence, intervene, and make recommendations? Several studies suggest that they are not. We have recent data that provide further evidence that future and current physicians may not be sufficiently competent in alcohol issues to counsel their patients about alcohol. Medical school curriculum and Continuing Medical Education on alcohol abuse and addiction should be required of all students and physicians so they can be best prepared to prevent problems and identify and treat those for whom prevention has failed. PMID- 15132340 TI - Physicians practicing in methadone treatment programs: who are they and what do they do? AB - OBJECTIVE: This study examines the characteristics and roles of physicians practicing in methadone maintenance treatment programs (MTPs). METHODS: Physicians and clinic directors at 172 MTPs in the United States completed surveys. MTPs were selected for study participation based on their locations (large urban, urban, or nonurban area) ownership status (for profit and non profit), and size (patient capacity of 1-100, 101-300, and 300+). Weighted data were analyzed with descriptive and multivariate methods. RESULTS. Physicians were primarily white males aged 45 or older; 44% had 10 or more years of experience working in methadone treatment. Physicians reported spending 26% of their time completing administrative tasks. Most reported that they determine dosing levels on an individual patient basis. Average maintenance dose was 69 mg/day. CONCLUSIONS: Physicians' treatment practices play a major role in overall treatment, treatment retention, and outcomes. Physicians at for-profit and large urban MTPs reported spending the most time in direct patient contact. PMID- 15132341 TI - Establishment of a multidisciplinary Health Evaluation and Linkage to Primary care (HELP) clinic in a detoxification unit. AB - We evaluated the feasibility of establishing a multidisciplinary Health Evaluation and Linkage to Primary care (HELP) clinic at an urban residential detoxification unit. Patients received a clinical evaluation and facilitated linkage to primary medical care including personalized referral, reminders, and appointment rescheduling. Of 235 adults reporting alcohol, cocaine or heroin as first or second drug of choice and without a primary care physician, 178 (76%) received a full HELP clinic evaluation, 35 (15%) some clinic components, and 7 (3%) only a primary care appointment. Of those with a full evaluation, 28% received pneumococcal vaccination, and most received health behavior counseling. Over the subsequent 2 years, 131 (60%) of the 220 patients whom had any contact with the HELP clinic had at least one primary care visit. A multidisciplinary health clinic to evaluate patients during detoxification is feasible and can link patients with substance dependence to primary medical care. PMID- 15132342 TI - Gender differences in outcomes in an HMO-based substance abuse treatment program. AB - This study examined gender differences in treatment outcomes and outcomes predictors among 155 men and 81 women attending a gender-sensitive substance abuse treatment program. Bivariate analyses indicated women improved more than men in social/family and daily functioning domains, but differences disappeared after controlling for baseline characteristics. Multivariate models predicting treatment outcomes revealed that, across Addiction Severity Index domains, outcomes for men were predicted primarily by mental health and medical conditions, severity of the substance abuse problem, and treatment com- pletion. For women, in addition to treatment completion, outcomes were more likely to be predicted by social, socio-demographic, and life-history characteristics. For abstinence outcomes, women who completed treatment were 9 times as likely to be abstinent at 7-month follow-up as other women; men who completed were 3 times more likely to be abstinent than other men. Women with more severe psychiatric status and those who felt their life was out of control were less likely to be abstinent, as were men who lived alone. Clinicians targeting such factors differentially for men and women may enhance the effectiveness of treatment. PMID- 15132343 TI - Concurrent alcohol and cocaine dependence impact on physical health among psychiatric patients. AB - The aim of this study was to examine the association between active, concomitant cocaine and alcohol dependence and the prevalence and patterns of comorbid physical disorders in a sample of substance abusing hospitalized psychiatric patients. Three groups of patients (concomitant cocaine and alcohol dependence (AD + CD) (N = 38), alcohol dependence (AD) only (N = 38), and cocaine dependence (CD) only (N = 25)) consecutively admitted to a psychiatric-substance abuse dual diagnosis unit were comparatively examined for the frequency of comorbid physical disorders diagnoses, including viral hepatitis, sexually transmitted diseases, and on liver function tests and electrocardiographic abnormalities. The results indicated that the concomitant alcohol and cocaine dependence group had higher rates of multiple physical disorders and also of multiple hepatitis infections than either the alcohol-only or the cocaine-only groups. PMID- 15132344 TI - Outcome variables for anorexic males and females one year after discharge from residential treatment. AB - The overall goal of this study was to evaluate the outcome of a residential program for eating disorders that uses a multidimensional approach to treatment. Patients were males and females admitted with a diagnosis of Anorexia Nervosa using DSM-IV criteria. A phone survey was developed by our staff and applied to patients 15-months post discharge. Responses were analyzed using paired t-test and multiple regression analysis. From discharge to follow-up, the females experienced an average weight gain of almost 7 lbs (P = 0.03) and the males experienced an average weight gain of 19 lbs (P = 0.025). Multiple regression analysis showed that a higher weight at contact date was associated with a higher weight at discharge, less fasting and the male gender. This kind of study helps us evaluate treatment outcome and identify key variables that predict changes in anorexics' body weight over time. PMID- 15132345 TI - Change in the pattern of illegal drug use in an inner city population over 50: an observational study. AB - Though alcohol and prescription drug abuse are well studied in older adults, there is little information regarding illicit drug use in older adults. We investigated illicit drug use in older adults presenting to an inner city emergency department in Los Angeles over a 10-month period. Of 3,417 adults over 50 presenting to the Emergency Department (ED), charts of 107, whose urine toxicology screens were positive for illicit drugs, were reviewed retrospectively. Cocaine was used most frequently (63%), followed by opiates (16%) and marijuana (14%). These rates of drug usage essentially mirror those seen in the general population of the Los Angeles area. Fifty-nine percent of patients using illicit drugs had cardiovascular disease compared with 10% of all older adults upon initial presentation to the ED. The use of illicit drugs by older persons may be an expanding phenomenon conferring serious but unrecognized health risks, and is in need of further study. PMID- 15132346 TI - Pre-treatment measures of impulsivity, aggression and sensation seeking are associated with treatment outcome for African-American cocaine-dependent patients. AB - We investigated whether measures of impulsivity, aggression and sensation seeking differed between cocaine-dependent subjects and controls, and whether these measures were related to treatment-outcome for cocaine patients. Pre-treatment assessments of impulsivity (Barratt Impulsivity Scale [BIS]), aggression (Buss Durkee Hostility Inventory [BDHI]) and sensation seeking (Zuckerman Sensation Seeking Scale [SSS]) were obtained for 141 African-American cocaine-dependent patients entering a 12-week, intensive outpatient treatment program and 60 controls. The outcome measures were number of negative urine drug screens, days in treatment, dropout rates and number of treatment sessions. Cocaine patients reported significantly higher scores on the SSS, the BIS and the BDHI than controls. Furthermore, the SSS scores showed a significantly negative correlation with days in treatment and negative urines, and a significant positive correlation with the dropout rate. The BIS and the BDHI scores were significantly associated with days in treatment and dropout rates respectively. A combination of the three variables contributed significantly toward predicting retention and abstinence. Higher levels of pretreatment impulsivity and aggression and sensation seeking seem to associated with poor treatment outcome for cocaine dependent patients receiving intensive outpatient treatment. Combining these behavioral measures with other clinical predictors may help in early identification of 'poor responders' who may benefit from additional or alternative treatment approaches. PMID- 15132347 TI - Obesity and vocational and avocational overload of the joint as risk factors for osteoarthritis. PMID- 15132348 TI - Do risk factors for incident hip osteoarthritis (OA) differ from those for progression of hip OA? PMID- 15132349 TI - Who gets osteoarthritis and why? PMID- 15132350 TI - Specific gene defects leading to osteoarthritis. PMID- 15132351 TI - How important are genetic factors in osteoarthritis? PMID- 15132353 TI - Radiographic issues in imaging the progression of hip and knee osteoarthritis. PMID- 15132352 TI - Markers of altered metabolism in osteoarthritis. PMID- 15132355 TI - Relationship between symptoms and structural change in osteoarthritis. what are the important targets for osteoarthritis therapy? PMID- 15132354 TI - Cartilage oligomeric matrix protein as a marker of osteoarthritis. PMID- 15132356 TI - Recent approaches to understanding osteoarthritis pain. PMID- 15132357 TI - Pushing the limits of patient-oriented outcome measurements in the search for disease modifying treatments for osteoarthritis. PMID- 15132358 TI - Outcome measures for clinical trials of disease modifying osteoarthritis drugs in patients with hip osteoarthritis. PMID- 15132359 TI - Inflammation in osteoarthritis. PMID- 15132360 TI - The importance of subchondral bone in the progression of osteoarthritis. PMID- 15132362 TI - The role of proprioceptive deficits, ligamentous laxity, and malalignment in development and progression of knee osteoarthritis. PMID- 15132361 TI - Impact of exercise on osteoarthritis outcomes. PMID- 15132363 TI - Properties of unipolar electrograms recorded with a multielectrode basket catheter. AB - In the past few years, clinical trials with multielectrode "basket" catheters have provided unique recordings from the endocardium of intact in-situ human hearts. Analysis of these recordings is difficult because unipolar electrograms obtained from a basket catheter in the blood-filled cavity differ from those obtained by other mapping techniques such as endocardial balloons used during antiarrhythmic surgery. We investigated these differences using basket catheter recordings obtained in isolated porcine and canine hearts that could be filled with perfusion fluid and evacuated at will. The results indicated that the differences between basket and balloon recordings are largely attributable to the presence and absence of blood. Activation maps obtained in the presence and absence of blood were usually similar and only differed in a minority of cases at sites in which electrograms revealed multiple deflections. In conclusion, unipolar mapping using a basket catheter can be used with confidence for the creation of activation maps if appropriate care is taken in the interpretation of fractionated electrograms. PMID- 15132364 TI - Reconstruction of the 12-lead electrocardiogram from reduced lead sets. AB - In clinical practice, continuous recording of all leads of the 12-lead electrocardiogram (ECG) is often not possible. We wanted to assess how well absent, noisy, or defective leads can be reconstructed from different lead subsets and how well lead reconstruction performs over time. A data set of 234 24 hour ECG recordings was divided into an equally sized training and test set. Precordial leads were systematically removed, and for all lead subsets including both limb leads and at least one precordial lead, the absent leads were reconstructed using general and patient-specific reconstruction templates. Reconstruction performance was measured by correlation between the original and reconstructed leads over the QRS and T waves, by average and maximum absolute ST differences, and by agreement when a clinical decision rule was applied. Reconstruction performance over time was evaluated at baseline, at 20 minutes, and 1, 6, 12 and 24 hours after the start of each recording. Reconstruction accuracy was high (correlation > or =0.932, average ST difference < or =30 microV, agreement > or =94.9%) with general reconstruction for lead sets with 1 or 2 precordial leads removed but was less satisfactory when more leads were missing. Patient-specific reconstruction performed well when up to 4 precordial leads were removed (correlation > or =0.967, average ST difference < or =26 microV, agreement > or =95.7%). Patient-specific reconstruction performance initially slightly decreased and then stabilized over time but remained much better than general reconstruction after 24 hours. Accurate reconstruction of the 12-lead ECG from lead subsets is possible over time. General reconstruction allows reconstruction of 1 or 2 precordial leads, whereas up to 4 leads can be reconstructed well using patient-specific reconstruction. PMID- 15132365 TI - Drug-induced cardiac toxicity: emphasizing the role of electrocardiography in clinical research and drug development. AB - This review describes the role of electrocardiography in clinical research and drug development, and addresses its utility in defining cardiac toxicity from noncardiac investigational drugs. Principles for designing electrocardiographic monitoring for cardiac safety in clinical trials are also reviewed. PMID- 15132366 TI - A definitive or thorough phase 1 QT ECG trial as a requirement for drug safety assessment. AB - Prolongation of the QT interval by noncardiac drugs is the commonest cause of drug delays in development, nonapprovals and withdrawal from after marketing. The new regulatory guidance issued by the FDA-Health Canada ECG Concept document, requires irrespective of preclinical cardiac findings a definitive or thorough Phase I trial for all bioactive agents powered to exclude a 5-ms QTc effect (upper confidence interval = 10 ms) since such resolution is usually not possible with the variability inherent in ECG data from the usual trials in the target population. To design a definitive QT trial attention must be given to the sources of QTc duration spontaneous variability. The sources include the proper selection of the sample size, frequency of and method to analyze ECGs, proper correction formula for QT duration, choice of the supratherapeutic dose (required since the trail must be conducted in healthy volunteers rather than the target population) and proper use of a placebo and positive control groups. The positive control group is essential to define the sensitivity of the trial to detect a drug's effect on cardiac repolarization. An approach to interpretation of the resulting ECG data from the trial is provided. PMID- 15132367 TI - Influence of sleep apnea on autonomic nervous activity and QT dispersion in patients with essential hypertension and old myocardial infarction. AB - Sleep apnea syndrome (SAS) is an important cardiovascular risk factor in patients with hypertension or myocardial infarction (MI). We evaluated the influence of SAS on autonomic nervous activity and QT dispersion in patients with hypertension or coronary artery disease with old MI. A portable sleep polygraph was attached to 30 healthy volunteers (N group), 30 patients with essential hypertension (HT group), and 30 patients with old myocardial infarction (MI group) to serially record oronasal respiration, tracheal sound, thoracic respiratory movement, and percutaneous arterial oxygen saturation. In addition, a digital Holter ECG was used to examine heart rate variability during nighttime sleep. Heart rate variability was analyzed by obtaining low-frequency (LF) power, high-frequency (HF) power, the LF/HF ratio, and very low-frequency (VLF) power. Dispersion of QT intervals was obtained by CM5 and CM1 leads. VLF and LF powers were significantly higher in the HT-SAS group (hypertensive patients with SAS) than the N and HT NSAS groups (hypertensive patients without SAS). The HF power was significantly lower in the HT-NSAS group than the N group, but the decrease in HF power in hypertension was not observed in the HT-SAS group. The LF/HF ratio was significantly higher in the HT-NSAS group than the N group, and this value was further increased in the HT-NSAS group. Percutaneous arterial oxygen saturation was decreased, and QT dispersion was significantly increased in the MI group during sleep apnea episodes. More severe autonomic nervous dysfunction and increased QTc dispersion were observed in hypertensive patients with SAS during episodes of apneas and hypopneas compared to those without SAS. These findings suggest that SAS may be associated with the future development of cardiac events. PMID- 15132368 TI - Effect of electrophysiologic character of ventricular premature beat on heart rate turbulence. AB - Heart rate turbulence (HRT) has been described as a predictor of high-risk patients with cardiac diseases. The purpose of this study is to determine how the degree of prematurity of a ventricular premature beat (VPB%) and retrograde ventriculoatrial (VA) conduction of VPBs affect HRT. We studied 30 patients without organic heart disease. We calculated turbulent slope (TS) and turbulent onset (TO) from VPBs induced by programmed stimulation from the right ventricular apex. TS was inversely and TO was positively correlated to VPB%. Without retrograde VA conduction of VPBs, TS was inversely and TO was positively correlated to VPB%. In VPBs with retrograde VA conduction, there were no significant correlations between TO and TS with VPB%. In conclusion, TS and TO calculated from VPBs with different degrees of prematurity varied widely. Both VPB% and characteristics of retrograde VA conduction may affect HRT. PMID- 15132369 TI - Characteristics of new-onset ventricular arrhythmias in pregnancy. AB - The characteristics and underlying mechanisms of new-onset ventricular arrhythmia during pregnancy have not been adequately investigated. We studied 11 pregnant women aged 35.4 +/- 3.1 years who experienced new-onset ventricular arrhythmias during pregnancy. The onset of their first episode of ventricular arrhythmia was distributed equally over the 3 trimesters. Ventricular premature contractions (VPCs) and ventricular tachycardia (VT) exhibited a monomorphic configuration in all patients; in 8 (73%) they originated from the right ventricular outflow tract. The frequency of VPCs decreased by more than 95% in 83% of the patients; couplets and VTs disappeared completely in all patients during the postpartum period. All parameters of heart rate variability were significantly lower during pregnancy than postpartum (P < .05). QT and QTc intervals were normal during pregnancy in all patients. The slope and the intersect of the QT/RR relationship obtained from Holter ECGs were not significantly different during pregnancy and in the postpartum period. New-onset ventricular arrhythmias in pregnancy manifest characteristics similar to those of idiopathic VT. Our findings suggest that various hemodynamic and neurohormonal changes associated with pregnancy play an important role in ventricular arrhythmogenesis. PMID- 15132370 TI - Predictors of in-hospital ventricular fibrillation or torsades de pointes in patients with acute symptomatic bradycardia. AB - Severe bradyarrythmias remain as an important cause for hospital urgent admission and these patients can suffer potentially lethal complications (such as ventricular fibrillation [VF] and torsades de pointes [TdP]) between hospital admission and final therapy. Incidence and predictors of these tachyarrhythmias have not been well established. We retrospectively studied all consecutive patients (N = 243, age 75 +/- 10 years; 47% men) admitted to the emergency department of a general hospital between January 1998 and July 2000 for symptomatic bradyarrhythmia. Concomitant therapy included diuretics (25%), digitalis (10%), beta-blockers (10%), amiodarone (2%), and verapamil or diltiazem (8%). Syncope was the most frequent symptom at admission (54%). The most prevalent inclusion bradyarrhythmia was > or =second-degree AV block (82%). Eleven patients (4.5%) presented VF or TdP. Univariate predictors for these complications were previous amiodarone or diuretic intake, presentation as syncope, low serum potassium level, and longer QTc at admission. Multivariate analysis with logistic regression showed only therapy with diuretics and/or amiodarone and QTc at admission as significant predictors for TdP or VF development. Incidence of VF or TdP in patients admitted for symptomatic bradyarrhythmia is relatively important. A prolonged QTc interval and/or therapy with amiodarone or diuretics can predict their presentation. PMID- 15132371 TI - Transient giant R-wave, right axis deviation, and intraventricular conduction delay during exercise treadmill testing: a case report. AB - A 53 year old man complained of chest pain during an exercise treadmill test. Electrocardiogram revealed transient giant R-wave, right-axis deviation, intraventricular conduction delay, and ST-segment elevation in the inferolateral leads. Subsequent coronary angiography showed an 80% lesion in mid part of a nondominant left circumflex artery, whereas the other coronary arteries had mild atherosclerosis only. Percutaneous coronary intervention and stenting was performed on the left circumflex artery lesion. A follow-up exercise thallium scan 3 months later still showed an intermediate-sized, mild reversible perfusion defect in the inferior and lateral wall but the giant R-wave ECG pattern was not inducible anymore. Restudy coronary angiography showed no in-stent restenosis, but there was disease progression in the midpart of the right coronary artery. The initial electrocardiographic pattern is typical of the "giant R-wave syndrome." Severe coronary spasm superimposed on the underlying mild atherosclerotic lesion of the right coronary artery is hypothesized to be the cause of the initial event. Ad hoc direct stenting was performed on the right coronary artery lesion. The patient remained symptom-free with a normal thallium scan 9 months later. PMID- 15132372 TI - Bundle branch block on alternate beats during atrial fibrillation. AB - This article reports a case of tachycardia-dependent right bundle branch block (RBBB) occurring during atrial fibrillation. In some sections of the recording, an alternans occurs between complexes with a complete RBBB pattern and complexes showing normal intraventricular conduction or incomplete RBBB. Alternans is frequently observed during phases of fast and nearly regular rhythm, but it occurs even in the presence of a markedly irregular ventricular response. The RBBB alternans associated with short and regular RR intervals is likely to represent a manifestation of 2:1 bundle branch supernormal conduction, whereas alternans occurring with irregular cycles expresses a complex interaction between the RR cycle length and some mechanisms affecting intraventricular conduction, such as tachycardia-dependent bundle branch block, supernormal conduction and concealed retrograde activation of the anterogradely blocked bundle branch (the so-called "linking" phenomenon). PMID- 15132373 TI - Radiation therapy-induced electrical reset of an implantable cardioverter defibrillator device located outside the irradiation field. AB - Given the overlap in risk factors for cardiac and malignant disease, some patients carrying implantable cardioverter defibrillators (ICDs) may require radiotherapy. Although pacemaker malfunction caused by radiotherapy is well known, few data exist on interactions between ICDs and radiation therapy. This report presents a patient with a single-chamber ICD and a history of bronchial carcinoma. During radiotherapy, electrical restart of the ICD device had occurred, although the device was not located inside the irradiation field. This case report highlights the diagnostic challenge represented by the environment of therapeutic radiation. PMID- 15132374 TI - Limbal stem cell transplantation. AB - The past two decades have witnessed remarkable progress in limbal stem cell transplantation. In addition to harvesting stem cells from a cadaver or a live related donor, it is now possible to cultivate limbal stem cells in vitro and then transplant them onto the recipient bed. A clear understanding of the basic disease pathology and a correct assessment of the extent of stem cell deficiency are essential. A holistic approach towards management of limbal stem cell deficiency is needed. This also includes management of the underlying systemic disease, ocular adnexal pathology and dry eye. Conjunctival limbal autografts from the healthy contralateral eye are performed for unilateral cases. In bilateral cases, tissue may be harvested from a cadaver or a living related donor; prolonged immunosuppression is needed to avoid allograft rejection in such cases. This review describes the surgical techniques, postoperative treatment regimes (including immunosuppression for allografts), the complications and their management. The short and long-term outcomes of the various modalities reported in the literature are also described. PMID- 15132375 TI - A comparative study of small incision trabeculectomy avoiding Tenon's capsule vis a-vis trabeculectomy with Mitomycin-C. AB - PURPOSE: To compare the results of small incision trabeculectomy avoiding Tenon's capsule (SIT) vis-a-vis intraoperative use of Mitomycin-C (MMC) in primary chronic angle closure glaucoma. METHODS: A controlled prospective study was conducted on 60 consecutive primary chronic angle closure glaucoma patients requiring glaucoma filtration surgery. Patients were divided into two groups, Group I (n = 30): those undergoing SIT and Group II (n = 30): those undergoing trabeculectomy with MMC. Patients were followed up serially for 24 months and their intraocular pressure (IOP) was monitored. Success was defined as IOP < or = 22 mm Hg with no additional anti-glaucoma medication or laser/surgical intervention. Success was also defined as a 30% reduction from the initial IOP at which optic disc cupping and/or visual field changes occurred. RESULTS: The final mean IOP with SIT was 16.80 +/- 4.20 mm Hg as against 17.84 +/- 3.80 mm Hg with trabeculectomy with MMC. Final success rate of 93.3% was obtained with SIT versus 90% with trabeculectomy with MMC. No major complications were seen with either procedure. CONCLUSION: Small incision trabeculectomy safely and effectively reduces the IOP in over 90% cases. The advantages of this procedure over trabeculectomy with MMC are its low cost, use of a small (2.5 mm) limbal incision which obviates the dissection of Tenon's capsule and absence of any major complication. PMID- 15132376 TI - Paediatric open globe injuries. Visual outcome and risk factors for endophthalmitis. AB - PURPOSE: To study the incidence of open globe injuries and the outcome in children, and to study the risk factors for post-traumatic endophthalmitis. METHODS: Paediatric patient population. Retrospective analysis of 72 consecutive cases of open globe injury over 3 years (January 1998 to December 2000). RESULTS: The cause of trauma was sports related (n = 18), home-made bow and arrow (n = 16), household kitchen injuries (n = 10), cracker injuries (n = 7) and other miscellaneous outdoor activities (n = 16). In 5 children the cause could not be ascertained. Visual acuity of > or = 3/60 in the injured eye at the last follow up examination was recorded in 37 of 70 patients (52.86%) whose visual acuity could be tested. The final visual acuity was significantly poorer in eyes where primary repair was delayed beyond 24 hours of injury (P < 0.05). Post- traumatic endophthalmitis developed in 39 of 72 (54.16%) eyes. Bow and arrow and household injuries (P < 0.5) and eyes in which primary repair was delayed beyond 24 hours of injury (P < 0.01) had a higher risk of endophthalmitis in univariate analysis. In multivariate analysis delayed repair was the only significant risk factor for the occurrence of endophthalmitis (P = 0.014). CONCLUSION: Delayed repair, bow and arrow injuries and household injuries were associated with significantly higher risk of endophthalmitis. The incidence of endophthalmitis can be reduced by early referral of trauma cases and parental supervision. PMID- 15132377 TI - Transpupillary thermotherapy in subfoveal choroidal neovascular membrane secondary to age-related macular degeneration. AB - PURPOSE: To report our initial experience in the treatment of subfoveal choroidal neovascular membrane, secondary to age-related macular degeneration (AMD) by transpupillary thermotherapy (TTT). METHODS: Fifty consecutive patients with subfoveal choroidal neovascularisation (CNV) secondary to AMD, were included in the study. The parameters assessed before the TTT were visual acuity by ETDRS chart, scotoma score by Amsler grid chart, reading speed, fundus examination by direct and indirect ophthalmoscope as well as +90 Diopter lens followed by digital fundus photography and fluorescein angiography (FA). RESULTS: The letter visual acuity improved or stabilized in 72% cases up to 12 weeks after TTT. Mean scotoma score decreased from a mean of 47.56, to 43.56 at 6 weeks and to 37 at 12 weeks. Mean reading speed increased from 27.04 words/minute at pretreatment to 34.52 words/minute at 6 weeks and 37.33 words/minute 12 weeks after TTT. CONCLUSION: TTT is not only a cheaper alternative to photodynamic therapy (PDT), but also is an efficacious tool in stabilisation or improvement of visual acuity in the management of subfoveal choroidal neovascular membrane due to AMD. PMID- 15132378 TI - Clinical and histopathological characteristics of uveal melanoma in Asian Indians. A study of 103 patients. AB - PURPOSE: To study the clinical and histopathological characteristics of uveal melanomas in Asian Indians and compare them with other ethnic groups. MATERIALS AND METHODS: One hundred and three enucleated eyes with clinical diagnosis of uveal melanoma were retrospectively studied (1987-2001) and the clinical and pathological features analysed. RESULTS: Uveal melanomas constituted 0.02% of outpatients in a referral eye hospital in India over a 12-year period. The patients were predominantly males. Uveal melanomas tended to occur in the younger age. The mean age was 45.7 +/- 14.2 years. Melanoma involved the choroid in 90 (87.37%) patients, and both choroid and ciliary body in 13 (12.62%) patients. Mean basal diameter of the tumour was 13.43 +/- 5.32 mm. Mixed cell type was more common. CONCLUSION: Uveal melanomas are rare in the Asian Indians compared to those in the West. They occur in younger persons, have a greater mean basal diameter than that of Caucasians and are predominantly of the mixed cell type. PMID- 15132379 TI - Transpupillary thermotherapy for choroidal neovascular membrane in age related macular degeneration. AB - PURPOSE: To evaluate the efficacy of transpupillary thermotherapy (TTT) in choroidal neovasularisation (CNVM) secondary to age related macular degeneration (AMD). MATERIAL AND METHODS: Retrospective, non-randomized study of 28 eyes of 28 patients with subfoveal CNVM (classic, occult or mixed) secondary to AMD. RESULTS: Fifteen patients (53.57%) maintained their pre-treatment vision, 2 (7.14%) patients showed improvement of more than 2 lines and 11 (39.28%) patients showed deterioration of vision by >2 lines. Angiographic and clinical regression of CNVM was noted in 19 patients (67.8%) on an average follow up of 15.32 +/- 3.31 months. CONCLUSION: TTT leads to stabilisation of vision in 60% of treated eyes with CNVM due to AMD. PMID- 15132380 TI - Elevated free tear lactoferrin levels in leprosy are associated with Type 2 reactions. AB - PURPOSE: To determine the association of demographics, leprosy and ocular characteristics with altered levels of lactoferrin in the tears of normal subjects and leprosy patients, and to detect the presence of antibodies to lactoferrin in these tear samples. METHOD: We collected light-stimulated tears from 298 leprosy patients and an equal number of normal subjects using the glass capillary method. Free lactoferrin levels were estimated using ELISA and the presence of antibodies to lactoferrin was detected using the immuno-blotting method. Significant associations were looked for between tear lactoferrin levels and demographic characteristics, leprosy characteristics such as type of disease, duration of disease, reactions, deformity and bacterial load, and ocular complications, using chi-square and regression analysis. RESULTS: Tear lactoferrin levels with a mean (SD) of 2.55 (2.83)mg/ml in the control group were significantly different (P<0.000) from leprosy patients with a smean (SD) of 5.66 (7.21)mg/ml. Age showed an inverse correlation with tear lactoferrin levels in controls. Increased bacterial load, grade 2 leg deformity and Type 2 reactions were significantly associated (P<0.05) with increased tear lactoferrin levels. Type 2 reactions remained significantly associated (P=0.01) on multiple regression analysis. Tear lactoferrin levels were not associated with gender, serum lactoferrin levels, Type 1 reactions, face patches, treatment status, orbicularis oculi weakness, lagophthalmos, ectropion, entropion, corneal opacity, cataract and iridocyclitis. CONCLUSION: Age is inversely related to tear lactoferrin levels in normal subjects. Free lactoferrin levels in tears are significantly higher in leprosy patients compared with normal controls. Type 2 reactions in leprosy are significantly associated with elevated tear lactoferrin levels. PMID- 15132381 TI - Intravitreal live Gnathostoma spinigerum. AB - Intraocular infestation by live Gnathostoma spinigerum is a rare occurrence in humans. Most of the published reports are from South-East Asia. We report a case of intravitreal gnathostomiasis, where the worm was removed live and intact by pars plana vitrectomy. PMID- 15132382 TI - Bilateral senile scleral plaques mimicking post-inflammatory scleral ectasia. AB - Scleral plaque is a commonly occurring change in older individuals. We report a case of bilateral scleral plaques seen in an elderly female patient. This current case report describes a common but often missed benign scleral change in the elderly individual. PMID- 15132383 TI - Seroprevalence of human immunodeficiency virus, hepatitis B virus and hepatitis C virus among eye donors. AB - Blood specimens collected at the time of enucleation of the eyes from 483 consecutive eye donors were tested for sero-markers of Human Immunodeficiency Virus (HIV), Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV). Antibodies to HIV1 were detected in 3 (0.62%), HBsAg in 17 (3.52%) and antibodies to HCV in 7 (1.45%). PMID- 15132384 TI - Locally aggressive orbital fibrous histiocytoma. AB - We report a rare presentation of a massive, locally aggressive, fibrous histiocytoma of the orbit. The importance of histopathological grading and appropriate management are highlighted. PMID- 15132385 TI - An unusual intraorbital foreign body. AB - A plastic foreign body penetrating the anterior base of skull through the orbit in a 10-year-old male child is reported. PMID- 15132386 TI - First contact management of postoperative endophthalmitis. A retrospective analysis. AB - Records of 37 consecutive patients of postoperative endophthalmitis referred to our centre from North India were retrospectively analysed to study the first contact management profile. Ten (27%) patients had received intravitreal antibiotics as a primary mode of treatment, and 27 (73%) had received only parenteral antibiotics. The outcome was worse in the latter group. This suggests that general ophthalmologists lack the capability to provide adequate treatment to patients with endopthalmitis in Northern India. PMID- 15132387 TI - Self retaining contact lens system for vitreous surgery. AB - We describe the principle and desiign of a new self-retaining contact lens system for vitreous surgery. The system has three lenses: theplano-concave, prism and magnifying lens. This system is based on the principle of a direct imaging contact lens, designed for a 150-200mm focal length operating micrcroscope. The contact lenses are designed to have an inferior concave surface [radius of curvature (ROC) 7.7mm], modified by theaddition of four footplates to provide stability and centration during vitreous surgery. The lenses are used with a drop of viscoelastic material placed between the concave surface of the contact lens and cornea. This induces negative suction and helps retain the lens in position during surgery. These specially designed lenses provide a stable, well-centered, high-resolution, magnified view of the fundus. This system eliminates the need for a skilled assistant or for suturing the lens to the sclera during vitreous surgery. PMID- 15132388 TI - Awareness of eye donation in the rural population of India. AB - PURPOSE: To determine the "awareness of eye donation" and "willingness to pledge eyes for donation" in the rural population of Andhra Pradesh, southern India. METHODS: A total of 7,775 subjects of all ages, representative of the rural population of Andhra Pradesh, participated in the Andhra Pradesh Eye Disease Study (APEDS). Subjects older than 15 years were interviewed regarding awareness of eye donation and willingness to pledge eyes for donation. RESULTS: Age-gender adjusted prevalence of awareness of eye donation in this population was 30.7% (95% CI: 29.5-31.9) but only 0.1% (age-gender adjusted prevalence) (95% CI: 0.05 0.25) had pledged eyes. On multivariate analysis the awareness of eye donation was significantly less in those subjects > or = 70 years old (OR 0.7; 95% CI: 0.6 0.8), illiterates (OR 0.2; 95% CI: 0.1-0.2), females (OR 0.8; 95% CI: 0.7-0.9), lower socioeconomic status group (OR 0.4; 95% CI: 0.4-0.5) and Christians (OR 0.2; 95% CI: 0.1-0.6). Media comprised the major source of information about eye donation. Of those aware of eye donation, 32.9% were willing to pledge eyes, and 50.6% needed more information to decide whether or not to pledge their eyes. CONCLUSIONS: There needs to be more transfer of knowledge if more eyes are to be pledged. One-third of those aware of eye donation have not pledged their eyes, and an additional 50.6% needed more information to decide. This means only about one-fifth of those aware of eye donations have pledged their eyes. PMID- 15132389 TI - Sparfloxacin corneal deposits. PMID- 15132390 TI - Debris in phacoemulsification handsets. A potential cause of endophthalmitis after cataract surgery? PMID- 15132391 TI - Essential parameters for accurate intraocular lens (IOL) power estimation in post refractive surgery cataract patients. PMID- 15132392 TI - Rhino-orbital-cerebral mucormycosis. A retrospective analysis and treatment option. PMID- 15132393 TI - Digital ophthalmic photography. PMID- 15132394 TI - Digital ophthalmic photography. PMID- 15132395 TI - Transpupillary thermo therapy for the treatment of choroidal neovascularisation secondary to age related macular degeneration in Indian eyes. PMID- 15132396 TI - Suture and surgically induced astigmatism after cataract surgery. PMID- 15132397 TI - Severe outbreak of Escherichia coli O157:H7 in health care institutions in Charlottetown, Prince Edward Island, fall, 2002. PMID- 15132398 TI - Adjuvant chemotherapy for early breast cancer: protocol and nonprotocol treatment options. Proceedings of a national research leader think tank and 3 regional breast cancer update working group meetings. July-October 2003. PMID- 15132399 TI - Alfred Hershey--Nobel Prize for work in virology. PMID- 15132400 TI - Pneumococcal bacteremia: lessons learned, yet more to learn. PMID- 15132401 TI - Changes in the epidemiology of pneumococcal bacteremia in a Swiss university hospital during a 15-year period, 1986-2000. AB - OBJECTIVE: To evaluate changes in epidemiological characteristics and outcome of patients with pneumococcal bacteremia during a 15-year period in a Swiss university hospital. PATIENTS AND METHODS: We reviewed the medical records of all hospitalized adults at the University Hospital Basel, Basel, Switzerland, whose blood culture yielded Streptococcus pneumoniae from January 1, 1986, through December 31, 2000. RESULTS: We analyzed 405 episodes of pneumococcal bacteremia in 394 patients. The mean annual incidence of 1.78 episodes per 1000 hospital admissions was inversely related to the mean atmospheric temperature of the area. During the study period, penicillin nonsusceptibility increased from 0% to 17%. The overall case-fatality rate was 25%, which decreased from 33% to 17% between the first and the second half of the study period (P<.001). The proportion of women with pneumococcal bacteremia increased from 37% to 52%. Independent risk factors for fatal outcome were coronary artery disease (P<.001; relative risk [RR], 4.3; 95% confidence interval [CI], 3.4-5.1), neutropenia (P=.001; RR, 3.2; 95% CI, 1.9-4.8), and age 65 years or older (P=.001; RR, 2.9; 95% CI, 1.8-4.2), whereas prior respiratory tract infection (P=.03; RR, 0.3; 95% CI, 0.1-0.5) and the occurrence of pneumococcal bacteremia in the second half of the study period (P=.01; RR, 0.4; 95% CI, 0.2-0.6) were independent predictors of survival. The case-fatality rate in human immunodeficiency virus (HIV)-infected patients was significantly lower than in patients not infected with HIV or in those with unknown HIV status (9% vs 27%; P=.006), which correlated with the younger mean +/ SD age of HIV-infected patients (33.2+/-6.6 years) compared with patients not infected with HIV (63.1+/-18.1 years) (P<.001). CONCLUSIONS: The case-fatality rate of patients with pneumococcal bacteremia decreased significantly between the first and second half of the study period, despite the increased prevalence of penicillin-nonsusceptible isolates. Independent risk factors for fatal outcome were coronary artery disease, neutropenia, and age 65 years or older, whereas prior respiratory tract infection and the occurrence of pneumococcal bacteremia in the second half of the study period were independent predictors of survival. HIV infection was a predisposing factor for pneumococcal bacteremia but was not a risk factor for fatal outcome. PMID- 15132402 TI - Outcomes and in-hospital treatment of out-of-hospital cardiac arrest patients resuscitated from ventricular fibrillation by early defibrillation. AB - OBJECTIVE: To describe and evaluate the in-hospital treatment of ventricular arrhythmias and underlying structural heart disease in patients who survive ventricular fibrillation (VF) out-of-hospital cardiac arrest (OHCA) in a region with a high survival rate after hospital discharge. PATIENTS AND METHODS: The study included all patients presenting in Olmsted County, Minnesota, who had experienced OHCA between November 1990 and December 2000 and who underwent defibrillation of VF by an emergency medical service system. RESULTS: Of 200 patients who experienced VF arrest, 138 (69%) survived to hospital admission (7 died in the emergency department before admission), and 79 (40%) were discharged. Of patients who were discharged, 37 (47%) had a reversible cause of the arrest (perimyocardial infarction) and received treatment of the primary process. The other 42 patients who were discharged had ischemic coronary heart disease (CHD) (n=25), nonischemic CHD (n=10), or idiopathic VF (n=7). Four of the patients with CHD but no left ventricular dysfunction were treated with coronary artery bypass grafting or percutaneous coronary intervention alone. A total of 52 patients (66%) were candidates for electrophysiologic testing. Of these patients, 48 (92%) underwent electrophysiologic testing; of these patients, 10 received amiodarone alone, and 35 received an implantable cardioverter-defibrillator (ICD) (of whom 3 also received amiodarone). Patients who did not receive ICD therapy typically presented before 1998 with CHD and underwent coronary artery bypass grafting or percutaneous coronary intervention only. Of 79 patients who were discharged, 14 (18%) with an ICD have received subsequent shocks. Nineteen (24%) of 79 patients have died, 5 of a primary cardiac etiology (including 2 with repeated OHCA). CONCLUSIONS: The VF OHCA survival rate is high in the setting of rapid defibrillation, with 40% of patients being discharged from the hospital. By the end of the 10-year study, more patients were receiving antiarrhythmic therapy, in particular ICD implantation, after hospital admission. Overall, the long-term survival in patients with VF OHCA is favorable. PMID- 15132403 TI - Efficacy and safety of ezetimibe coadministered with simvastatin in patients with primary hypercholesterolemia: a randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To compare the efficacy and safety of 10 mg of ezetimibe coadministered with simvastatin with the safety and efficacy of simvastatin monotherapy for patients with hypercholesterolemia. PATIENTS AND METHODS: This multicenter double-blind, placebo-controlled, factorial study enrolled 887 patients with hypercholesterolemia (low-density lipoprotein cholesterol [LDL-C], 145-250 mg/dL; triglycerides, < or = 350 mg/dL). Patients were randomized to 1 of 10 treatments--placebo, ezetimibe at 10 mg/d, simvastatin at 10, 20, 40, or 80 mg/d, or simvastatin at 10, 20, 40, or 80 mg/d plus ezetimibe at 10 mg/d for 12 weeks. The study began March 13, 2001, and ended January 8, 2002. The primary efficacy end point was the mean percent change in LDL-C levels from baseline to study end point (last available postbaseline LDL-C measurement) for the pooled ezetimibe/simvastatin group vs the pooled simvastatin monotherapy group. RESULTS: Coadministration of ezetimibe/simvastatin was significantly (P<.001) more effective than simvastatin alone in reducing LDL-C levels for the pooled ezetimibe/simvastatin vs pooled simvastatin analysis and at each specific dose comparison. The decrease in LDL-C levels with coadministration of ezetimibe and the lowest dose of simvastatin, 10 mg, was similar to the decrease with the maximum dose of simvastatin, 80 mg. A significantly (P<.001) greater proportion of patients in the ezetimibe/simvastatin group achieved target LDL-C levels compared with those in the monotherapy group. Treatment with ezetimibe/simvastatin also led to greater reductions in total cholesterol, triglyceride, non-high-density lipoprotein cholesterol, and apolipoprotein B levels compared with simvastatin alone; both treatments increased high-density lipoprotein cholesterol levels similarly. The safety and tolerability profiles for the ezetimibe/simvastatin and monotherapy groups were similar. CONCLUSION: Through dual inhibition of cholesterol absorption and synthesis, coadministration of ezetimibe/simvastatin offers a highly efficacious and well-tolerated lipid lowering strategy for treating patients with primary hypercholesterolemia. PMID- 15132404 TI - Clinical characteristics and mechanisms of stroke after polytrauma. AB - OBJECTIVES: To describe the clinical characteristics of acute stroke attributable to trauma and to analyze the mechanisms and effect of stroke on outcome in patients with polytrauma. PATIENTS AND METHODS: We retrospectively reviewed the medical records of patients with polytrauma complicated by stroke at the Mayo Clinic in Rochester, Minn. The diagnosis of stroke was matched with trauma for the period between January 1, 1985, and December 31, 2001. Polytrauma was defined as at least 2 injuries that involve at least 1 vital organ (eg, lung or liver) and necessitate patient admission to a trauma intensive care unit. RESULTS: We identified 14 patients with polytrauma who had experienced a stroke (13 ischemic and 1 hemorrhagic). Craniocervical artery dissection was the most common mechanism (7 patients), and skull or facial fractures were a possible marker for this (5 of 7 patients). There were frequent delays in identifying stroke, with limb fracture on the side of the paresis (5 of 6 patients) being a possible factor. Four patients died of postischemic brain swelling, and 5 had major neurologic impairment. CONCLUSIONS: Stroke after polytrauma is due to various mechanisms, is frequently difficult to recognize early, and profoundly affects patient outcome. PMID- 15132405 TI - Site specificity of regular health club exercise on muscle strength, fitness, and bone density in women aged 29 to 45 years. AB - OBJECTIVE: To determine the effect of regular exercise with use of stepper or skier machines on muscle strength, cardiovascular fitness, bone mineral density, and markers of bone turnover in women aged 29 to 45 years. SUBJECTS AND METHODS: We evaluated 14 women: 5 who used a stepper machine for at least 3 hours each week for 2 years, 4 who used a skier machine for at least 3 hours each week for 2 years, and 5 who did not exercise (controls). All women were healthy, had no history of disease or medication use known to affect bone metabolism, and had normal levels of estradiol-17beta. Differences between the exercise and control groups were analyzed with the Kruskal-Wallis test. RESULTS: No significant differences were noted between the exercise groups and the control group with respect to age, height, total body bone mineral density, calcium intake, and metabolic bone markers. Significant differences were noted between the groups in body weight (stepper vs control, P=.03; skier vs control, P=.02), body mass index (stepper vs control, P=.03; skier vs control, P=.02), Physical Activity Score (stepper vs control, P=.009; skier vs control, P=.01), percentage body fat (stepper vs control, P=.03; skier vs control, P=.02), and maximum oxygen consumption (stepper vs control, P=.009; skier vs control, P=.009). Bilateral hip extensor strength was significantly stronger in the stepper group than in the skier group (P=.03). Grip strength was significantly stronger in the stepper group than in the skier group (right, P=.01; left, P=.049). CONCLUSION: Although comparable health club exercises can contribute to fitness, their site-specific effect varies. Stepper machines are preferred over skier machines for improving hip extensor strength. PMID- 15132406 TI - Primer on medical genomics. Part XIII: Ethical and regulatory issues. AB - Ethics in the new genomics era has become an increasingly complex subject that often arouses passion and confusion. Although 50 years have elapsed since the elucidation of the DNA molecule, the recent near-complete sequencing of the human genome has sharply accelerated the incorporation of genetics into the medical mainstream. Along with these scientific advances, however, have surfaced challenges, liabilities, and issues regarding the processing and management of genetic information as they relate to core ethical principles such as respect for autonomy, beneficence, nonmaleficence, and justice. Institutions and state and federal governments have initiated systematic and preemptive measures in education, resource development, and protective legislation to address these cardinal ethical issues. Genetic research is also being scrutinized carefully by institutional review boards, an activity that should not be perceived as being adversarial but rather as a protective shield for investigators and research participants alike. Ultimately, it is hoped that genomics medicine will diminish rather than enhance existing sex-, race-, and socioeconomic class-based inequities in health care access and delivery. This article describes some but not all aspects of the ethical, legal, and social implications of genomics in clinical practice. PMID- 15132407 TI - Primer on medical genomics. Part XIV: Introduction to systems biology--a new approach to understanding disease and treatment. AB - The advent of the "-omics revolution" has forced us to reevaluate our ability to acquire, measure, and handle large data sets. Omic platforms such as expression arrays and mass spectrometry, with their exquisite selectivity, sensitivity, and specificity, are unrivaled technologies for detection, quantitation, and identification of DNA, messenger RNA, proteins, and metabolites derived from complex body tissue and fluids. More recently, attempts have been made to capture the utility of these platform technologies and combine them under the umbrella of systems biology, also referred to as pathway, network, or integrative biology. Applied systems biology is the integrated analysis of genetic, genomic, protein, metabolite, cellular, and pathway events that are in flux and interdependent. It necessitates the use of a variety of analytic platforms as well as biostatistics, bioinformatics, data integration, computational biology, modeling, and knowledge assembly protocols. Such sophisticated analyses may provide new insight into the understanding of disease processes and mechanisms of action of pharmaceutical agents. Ultimately, this requires a perspective on how complex systems behave and are modulated. In this regard, systems biology, more appropriately considered as a process containing a series of modules, aims to provide tools and capabilities to carry out such tasks. We describe the essentials required to carry out systems biology experiments, the method in which integrated data in the form of a systems biology correlation network affords new insight into understanding disease, and the vista of developing more efficient biomarkers and therapeutic agents. PMID- 15132408 TI - Primer on medical genomics. The end of the beginning. PMID- 15132410 TI - Unappreciated prevalence of interatrial block and associated consequences: a poorly perceived pandemic. PMID- 15132409 TI - Relationship of treatment-resistant head lice to the safety and efficacy of pediculicides. AB - Head lice infestation is a common and growing problem that primarily affects school-aged children. Most cases of head lice are diagnosed and treated by nonphysicians. Misdiagnosis may lead to treatment when no lice are present. Treatment failure may lead to repeated use of and improperly applied pediculicides, potentially resulting in overexposure to pesticides. These treatment failures are primarily due to the emergence of treatment-resistant lice. In regions where resistant lice are common, patients may self-treat numerous times with over-the-counter pediculicides before seeking treatment from a physician. Resistance has decreased the efficacy of lindane, a prescription pediculicide that has been used for decades. In addition, the Food and Drug Administration recently warned of potentially serious adverse effects associated with lindane and recommended strict controls for its use. Malathion, recently reintroduced in the United States as a prescription pediculicide, has not been associated with treatment resistance or notable adverse effects, although it is flammable due to its alcohol base. Because of concerns about decreasing efficacy due to resistance and safety concerns about over-the-counter products and some prescription pediculicides, a reassessment of pediculicide safety is warranted. The safety and efficacy of commonly used over-the-counter and prescription pediculicide products are discussed, along with the safety and efficacy of other treatments, such as ivermectin, that are not indicated for the treatment of head lice but are being used increasingly. PMID- 15132411 TI - Overview, prevention, and treatment of rabies. AB - Rabies is a uniformly fatal viral encephalitis that causes 30,000 to 70,000 deaths worldwide each year. Prevention is the primary approach to the disease. In the United States, 25,000 to 40,000 people are treated annually for exposure to rabid or potentially rabid animals at a per-patient cost exceeding 1000 dollars. Rabies is transmitted usually by saliva from infected animal bites. However, recent findings that rabies can be transmitted from bats to humans by relatively casual contact has resulted in dramatic changes in guidelines from the Centers for Disease Control and Prevention for postexposure prophylaxis. We review the 5 clinical stages of rabies, current methods of diagnosis, and prevention in animal reservoirs and in humans. We also discuss the use of rabies immune globulin and active and passive vaccinations for preexposure prophylaxis and postexposure treatment of rabies. Human exposure to rabies will always be a possibility, but methods to prevent the disease both before and after exposure to the virus are safe and readily available. PMID- 15132412 TI - 32-year-old man with episodic weakness. PMID- 15132413 TI - Bilateral carotid body paraganglioma. PMID- 15132414 TI - Epidural abscess and meningitis after epidural corticosteroid injection. AB - Epidural abscess with and without associated meningitis after epidural corticosteroid injections for radicular back pain is a rarely reported complication. We report the occurrence of an epidural abscess and meningitis in a 70-year-old man after 2 epidural corticosteroid injections for treatment of acute radicular lumbar back pain. At the time of diagnosis, cerebrospinal fluid cultures grew Staphylococcus aureus, and the patient was treated with intravenous antibiotics. Possible predisposing factors for the development of an epidural abscess and meningitis in this patient include a 2-year history of neutropenia and an accidental dural puncture that occurred during performance of the first epidural injection. A literature search identified 11 reported cases of epidural abscess, 2 of epidural abscess and meningitis, and 1 of meningitis attributed to epidural corticosteroid injections. Eight of the 14 reported patients were immunocompromised, and 8 (67%) of the 12 in whom cultures of blood, cerebrospinal fluid, or epidural pus were performed had results positive for S. aureus. Antibiotic prophylaxis for S. aureus should be considered for immunocompromised patients undergoing epidural corticosteroid injections. PMID- 15132415 TI - Severe, reversible pulmonary hypertension in a patient with monoclonal gammopathy and features of dermatomyositis. AB - Pulmonary hypertension is an extremely serious and potentially fatal disorder. Although pulmonary hypertension is a potential complication of connective tissue disease, it has been reported rarely in patients with dermatomyositis. Similarly, multiple myeloma is rare in patients with dermatomyositis. We describe a patient with severe pulmonary hypertension who also had features of dermatomyositis and monoclonal gammopathy. To our knowledge, this is the first reported case of a patient in whom all 3 disorders occurred concurrently. Even more striking is the fact that the patient responded to treatment with cyclophosphamide and prostacyclin. He is asymptomatic more than 5 years after treatment was discontinued. PMID- 15132416 TI - Successful treatment of symptomatic coronary endothelial dysfunction with enhanced external counterpulsation. AB - Enhanced external counterpulsation (EECP) is a valuable therapeutic option for patients with coronary artery disease and refractory angina. Although the exact mechanisms by which this technique exerts favorable effects remain unclear, improvement in endothelial function is considered a potential mechanism contributing to the clinical benefit associated with EECP. We describe a young woman with severely symptomatic coronary endothelial dysfunction in the absence of obstructive coronary artery disease who experienced a dramatic and sustained reduction in symptoms in response to a standard 35-hour course of EECP. PMID- 15132417 TI - The physiology and treatment of vitamin D deficiency. PMID- 15132418 TI - Vitamin D deficiency and chronic pain: cause and effect or epiphenomenon? PMID- 15132419 TI - Vitamin D deficiency as a contributor to multiple forms of chronic pain. PMID- 15132420 TI - Sun exposure, vitamin D metabolism, and skin cancer. PMID- 15132421 TI - Ocular ethambutol toxicity. PMID- 15132422 TI - The hunter-gatherer diet. PMID- 15132423 TI - Successful treatment of chronic meningitis caused by Scedosporium apiospermum with oral voriconazole. PMID- 15132424 TI - Two causes of hypercalcemia: learning by the Holmesian method. PMID- 15132425 TI - A 1921 letter from Vermont with a tribute to the Charles H. Mayo family. PMID- 15132426 TI - Hippocampal prosaposin changes during stress: a glucocorticoid-independent event. AB - Several studies indicate that stress can produce remarkable effects on neurotrophic factors. In this regard, hippocampus is the most interesting structure of the brain because of its broad involvement in behavioral and neuroendocrine phenomena. In the present study, we investigated the effect of stress on hippocampal prosaposin, which is known to act as a neurotrophic and neuroprotective factor. Rats subjected to restraint stress (120 min) had a significant and transient reduction of hippocampal, but not hypothalamic, prosaposin full-length protein. Indeed, when this stressful stimulus was applied daily for 3 days, no differences were detected in comparison with naive rats. To investigate the role of glucocorticoids in the stress-induced decrease in hippocampal prosaposin, adrenalectomized and corticosterone-treated rats were studied. The results indicate that adrenalectomized rats behave as intact animals. This finding indicates that the absence of endogenous corticosterone does not prevent a decrease in hippocampal prosaposin. When an increase of corticosterone was achieved through exogenous administration, hippocampal prosaposin concentrations were unchanged in comparison with vehicle-injected (sesame oil) rats. These results led to the conclusion that stress, not via an increase of glucocorticoid hormone, transiently reduces hippocampal prosaposin levels. This phenomenon is followed by rapid recovery of the neurotrophin level, even when the stress stimulus persists. PMID- 15132427 TI - Stress generates emotional memories and retrograde amnesia by inducing an endogenous form of hippocampal LTP. AB - Models of the neurobiology of memory have been based on the idea that information is stored as distributed patterns of altered synaptic weights in neuronal networks. Accordingly, studies have shown that post-training treatments that alter synaptic weights, such as the induction of long-term potentiation (LTP), can interfere with retrieval. In these studies, LTP induction has been relegated to the status of a methodological procedure that serves the sole purpose of disturbing synaptic activity in order to impair memory. This perspective has been expressed, for example, by Martin and Morris (2002: Hippocampus 12:609-636), who noted that post-training LTP impairs memory by adding "behaviorally meaningless" noise to hippocampal neural networks. However, if LTP truly is a memory storage mechanism, its induction should represent more than just a means with which to disrupt memory. Since LTP induction produces retrograde amnesia, the formation of a new memory should also produce retrograde amnesia. In the present report, we suggest that one type of learning experience, the storage of fear-related (i.e., stressful) memories, is consistent with this prediction. Studies have shown that stress produces potent effects on hippocampal physiology, generates long-lasting memories, and induces retrograde amnesia, all through mechanisms in common with LTP. Based on these findings, we have developed the hypothesis that a stressful experience generates an endogenous form of hippocampal LTP that substitutes a new memory representation for preexisting representations. In summary, our hypothesis implicates the induction of endogenous synaptic plasticity by stress in the formation of emotional memories and in retrograde amnesia. PMID- 15132428 TI - Hippocampal function in posttraumatic stress disorder. AB - Recent studies have reported memory deficits and reduced hippocampal volumes in posttraumatic stress disorder (PTSD). The goal of the current research was to use functional neuroimaging and a validated explicit memory paradigm to examine hippocampal function in PTSD. We used positron emission tomography (PET) and a word-stem completion task to study regional cerebral blood flow (rCBF) in the hippocampus in 16 firefighters: 8 with PTSD (PTSD group) and 8 without PTSD (Control group). During PET scanning, participants viewed three-letter word stems on a computer screen and completed each stem with a word they had previously encoded either deeply (High Recall condition) or shallowly (Low Recall condition). Relative to the Control group, the PTSD group exhibited significantly smaller rCBF increases in the left hippocampus in the High vs Low Recall comparison. However, this finding reflected relatively elevated rCBF in the Low Recall condition in the PTSD group. Collapsing across High and Low Recall conditions, (1) the PTSD group had higher rCBF in bilateral hippocampus and left amygdala than the Control group, and (2) within the PTSD group, symptom severity was positively associated with rCBF in hippocampus and parahippocampal gyrus. The groups did not significantly differ with regard to accuracy scores on the word stem completion task. The PTSD group had significantly smaller right (and a trend for smaller left) hippocampal volumes than the Control group. The results suggest an abnormal rCBF response in the hippocampus during explicit recollection of nonemotional material in firefighters with PTSD, and that this abnormal response appears to be driven by relatively elevated hippocampal rCBF in the comparison condition. PMID- 15132429 TI - Differential contributions of dorsal hippocampal subregions to memory acquisition and retrieval in contextual fear-conditioning. AB - The hippocampus is an essential neural structure in developing contextual memory in a situation in which rapid development of associative learning should occur. We tested a subregion-specific contribution in the hippocampus to memory acquisition and retrieval, using the contextual fear-conditioning paradigm. The current results suggest that all three subregions (i.e., CA3, CA1, and dentate gyrus) of the hippocampus contribute to rapid acquisition of contextual memory in the initial phase of acquisition. The involvement of CA3 seems to be important at the earliest stage of acquisition, presumably for developing instant representation of a context. The role of CA3, however, was minimal in retrieving contextual memory after a long time period (i.e., 24 h), whereas the other subregions (i.e., CA1 and dentate gyrus) were critically involved. The results indicate time-dependent differential contributions of the hippocampal subregions to memory acquisition and retrieval in contextual fear-conditioning. PMID- 15132430 TI - Amygdala-kindling induces alterations in neuronal density and in density of degenerated fibers. AB - Kindling is characterized by a progressive intensification of seizure activity culminating in generalized seizures following repeated administration of an initially subconvulsive electrical or chemical stimulus. Since it is known that epilepsy induces morphological alterations in the limbic system, we examined the neuropathological consequences of kindling with a sensitive silver-staining method for the visualization of damaged neurons and Nissl staining for the estimation of the neuronal densities in different limbic areas. Wistar rats implanted with electrodes in the left basolateral nucleus were stimulated until 15 consecutive stage V seizures (scale of Racine). Amygdala-kindled animals had reduced cell density in the amygdala and increased density of fragments of degenerated axons. Reduced neuronal density and the occurrence of degenerated axons in kindled animals were more prominent in the ipsilateral than in the contralateral hemisphere. In addition, more degenerated axons were found in cortical structures of kindled than sham-operated animals. These results indicate that kindling induced morphological alterations that were not restricted to either the ipsilateral hemisphere or the stimulated region. These morphological changes might be responsible for the emotional and behavioral disturbances that can accompany epilepsy. PMID- 15132431 TI - Postsynaptic injection of calcium-independent phospholipase A2 inhibitors selectively increases AMPA receptor-mediated synaptic transmission. AB - The calcium-independent form of phospholipase A2 (iPLA2), an enzyme known to generate arachidonic acid (AA), was recently identified as the predominant constitutive phospholipase in the hippocampus. The present study shows that the iPLA2 inhibitor bromoenol lactone, when introduced into hippocampal CA1 pyramidal cells through a patch pipette, generated a dose-dependent increase in the amplitude of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) receptor mediated excitatory postsynaptic currents (EPSCs). The iPLA2 inhibitor by itself interfered with neither paired pulse facilitation nor N-methyl-D-aspartate (NMDA) receptor-mediated EPSCs, suggesting that its influence on synaptic transmission is postsynaptic in origin and specific to the AMPA subtype of glutamate receptors. Comparable results were obtained with palmitoyl trifluoromethyl ketone, a second structurally distinct iPLA2 inhibitor. The ability of iPLA2 inhibitors to increase AMPA receptor-mediated currents was also reproduced by MK 866, an inhibitor recognized to interfere with the generation of 5-lipoxygenase by-products of AA. At the biochemical level, we found that AMPA, but not NMDA glutamate receptor subunits, were upregulated in rat brain sections pre-incubated with the iPLA2 inhibitors. Collectively, these results provide the first experimental evidence that constitutive iPLA2 and/or its metabolites play an important role in the postsynaptic modulation of neurotransmission in CA1 pyramidal cells of the hippocampus. PMID- 15132432 TI - Expression, interaction, and proteolysis of death-associated protein kinase and p53 within vulnerable and resistant hippocampal subfields following seizures. AB - Death-associated protein (DAP) kinase is a novel regulator of cell death whose in vivo target(s) and role in neuronal cell death remain uncertain. Since DAP kinase has been implicated in p53-mediated apoptosis, a pathway activated following epileptic brain injury, we examined the relationship between DAP kinase and p53 following seizures. Rats underwent brief (40-min) seizures evoked by intraamygdala kainic acid, which caused the death of ipsilateral CA3 neurons while preserving the contralateral CA3 subfield. Seizures caused a small decline in levels of the approximately 160-kD DAP kinase within injured ipsilateral hippocampus, commensurate with the appearance of an approximately 60-kD fragment, and proteolysis of the p53 inhibitor, murine double minute gene 2 (MDM2). Expression of p53 increased within the ipsilateral hippocampus, and DAP kinase was detected within p53 immunoprecipitates. In contrast, DAP kinase and MDM2 were not proteolyzed within the seizure damage-resistant contralateral hippocampus. Furthermore, DAP kinase and p53 did not interact within the contralateral hippocampus, and p53 cellular localization redistributed from the nucleus to cytoplasm commensurate with p53 proteolysis. These data suggest that DAP kinase may be involved in the p53 pathway during seizure-induced neuronal death. PMID- 15132433 TI - Neuregulin1 downregulates postsynaptic GABAA receptors at the hippocampal inhibitory synapse. AB - The growth factor neuregulin 1 (NRG1) has been proposed to contribute to the formation and maturation of neuromuscular and interneuronal synapses by upregulating the expression of specific neurotransmitter receptor subunits. In the present report, we show that, in the hippocampus, NRG1 is expressed in a pattern suggesting that it regulates synapse development in the CA1 region. However, in contrast to what has been shown in other synapses, NRG1 reduces the expression of gamma-aminobutyric acid (GABA)A receptors alpha subunits in hippocampal slices, and the mean amplitude of GABAergic miniature inhibitory postsynaptic currents (IPSCs) in hippocampal CA1 pyramidal neurons, without affecting IPSC kinetics or frequency. These effects of NRG1 occur without concomitant changes in glutamate receptors and other synaptic proteins. We propose that the role of NRG1 in the formation and maturation in the hippocampal inhibitory synapse is downregulation, rather than upregulation, of receptor subunit expression. These results suggest that NRG1 may contribute to the reduction in GABAergic synaptic activity in hippocampal CA1 pyramidal neurons that normally occurs during early postnatal development, and that alterations in NRG1 signaling in the hippocampus may contribute to schizophrenia and epilepsy. PMID- 15132434 TI - Ontogeny of seizure-induced increases in BDNF immunoreactivity and TrkB receptor activation in rat hippocampus. AB - The present work tested the hypothesis that the anatomic and developmental patterns of status epilepticus-induced increases of brain-derived neurotrophic factor (BDNF) protein coincided with status epilepticus-induced increases of phospho-Trk immunoreactivity, a measure of TrkB receptor activation, in rat hippocampus. In P22 rats, robust increases of phospho-Trk immunoreactivity were detected in the mossy fiber pathway of the hippocampus one day following kainate induced status epilepticus. Conversely, no change in phospho-Trk immunoreactivity was detected in P8 or P14 rats. In P17 rats, intermediate levels of increased phospho-Trk immunoreactivity were detected, again in the mossy fiber pathway. Like phospho-Trk immunoreactivity, marked increases of BDNF immunoreactivity were detected in the mossy fiber pathway of P22 but not P14 rats. Dissociations were found in P17 rats following status epilepticus in that striking increases of BDNF, but not phospho-Trk immunoreactivity were detected. Immunoprecipitation and Western blot analyses of hippocampal extracts after status epilepticus showed increased phospho-TrkB, but not TrkB immunoreactivity in P22 rats, thereby confirming and extending the immunohistochemical findings. While most of the findings support the hypothesis, important dissociations among individual animals at P17 were identified. Together the findings are consistent with the proposal that status epilepticus-induced increase of BDNF content in the mossy fibers is necessary, but not sufficient, to effect activation of TrkB, as revealed by phospho-Trk immunoreactivity. Furthermore, these results provide the first characterization of seizure-induced increases in BDNF protein and TrkB receptor activation in developing animals. PMID- 15132436 TI - Ionic mechanisms in the generation of subthreshold oscillations and action potential clustering in entorhinal layer II stellate neurons. AB - A multicompartmental biophysical model of entorhinal cortex layer II stellate cells was developed to analyze the ionic basis of physiological properties, such as subthreshold membrane potential oscillations, action potential clustering, and the medium afterhyperpolarization. In particular, the simulation illustrates the interaction of the persistent sodium current (I(Nap)) and the hyperpolarization activated inward current (Ih) in the generation of subthreshold membrane potential oscillations. The potential role of Ih in contributing to the medium hyperpolarization (mAHP) and rebound spiking was studied. The role of Ih and the slow calcium-activated potassium current Ikappa(AHP) in action potential clustering was also studied. Representations of Ih and I(Nap) were developed with parameters based on voltage-clamp data from whole-cell patch and single channel recordings of stellate cells (Dickson et al., J Neurophysiol 83:2562-2579, 2000; Magistretti and Alonso, J Gen Physiol 114:491-509, 1999; Magistretti et al., J Physiol 521:629-636, 1999a; J Neurosci 19:7334-7341, 1999b). These currents interacted to generate robust subthreshold membrane potentials with amplitude and frequency corresponding to data observed in the whole cell patch recordings. The model was also able to account for effects of pharmacological manipulations, including blockade of Ih with ZD7288, partial blockade with cesium, and the influence of barium on oscillations. In a model with a wider range of currents, the transition from oscillations to single spiking, to spike clustering, and finally tonic firing could be replicated. In agreement with experiment, blockade of calcium channels in the model strongly reduced clustering. In the voltage interval during which no data are available, the model predicts that the slow component of Ih does not follow the fast component down to very short time constants. The model also predicts that the fast component of Ih is responsible for the involvement in the generation of subthreshold oscillations, and the slow component dominates in the generation of spike clusters. PMID- 15132437 TI - A novel population of calretinin-positive neurons comprises reelin-positive Cajal Retzius cells in the hippocampal formation of the adult domestic pig. AB - Calretinin-containing neurons in the hippocampal formation, including the subiculum, presubiculum, parasubiculum, and entorhinal cortex, were visualized with immunocytochemistry. Calretinin immunoreactivity was present exclusively in non-principal cells. The largest immunoreactive cell population was found in the outer half of the molecular layer of the dentate gyrus and in the stratum lacunosum-moleculare of Ammon's horn. A proportion of these cells were also immunoreactive for reelin, a Cajal-Retzius cell marker. Similar calretinin positive cells were found in the molecular layer of the subicular complex and entorhinal cortex. In the parasubiculum, a few immunoreactive bipolar and multipolar cells could be observed in the superficial and deep pyramidal cell layers. In the entorhinal cortex, bipolar and multipolar calretinin-positive cells were frequent in layer II, and large numbers of multipolar cells in layer V were immunoreactive. Electron microscopic analysis showed that somata of calretinin-positive cells contained either round nuclei with smooth nuclear envelopes or nuclei with multiple deep infoldings. Immunoreactive dendrites were smooth varicose, and the apposing axon terminals formed both symmetric and asymmetric synapses. Zonula adherentia were observed between calretinin-positive dendrites. Calretinin-positive axon terminals formed two types of synapses. Axon terminals with asymmetric synapses were found close to the hippocampal fissure, whereas axon terminals forming symmetric synapses innervated spiny dendrites in both the molecular layer of the dentate gyrus and in stratum lacunosum-moleculare of Ammon's horn. Calretinin-positive axon terminals formed both symmetric and asymmetric synapses with calretinin-positive dendrites. In conclusion, calretinin positive neurons form two major subpopulations in the adult domestic pig hippocampus: (1) a gamma-aminobutyric acid (GABA)ergic subpopulation of local circuit neurons that innervates distal dendrites of principal cells in both the dentate gyrus and in Ammon's horn; and (2) Cajal-Retzius type cells close to the hippocampal fissure, as well as in the molecular layer of the subicular complex and entorhinal cortex. PMID- 15132435 TI - P/Q Ca2+ channel blockade stops spreading depression and related pyramidal neuronal Ca2+ rise in hippocampal organ culture. AB - Ca2+ channels and pyramidal cell Ca2+ are involved in hippocampal spreading depression (SD), but their roles remain elusive. Accordingly, we characterized Ca2+ changes during SD in CA3 pyramidal neurons and determined whether Ca2+ channel antagonists could prevent SD. SD was induced in hippocampal organotypic cultures (HOTCs), in which experimental conditions can be rigorously controlled. SD was triggered by transient exposure to sodium acetate (NaAc)-based Ringer's coupled to an electrical pulse in the dentate gyrus and its occurrence confirmed with interstitial DC recordings. Pyramidal cell Ca2+ was measured with fura-2 filled cells and was quantified at the soma, proximal and more distal apical dendrites. Regional Ca2+ changes began simultaneously with the triggering pulse of SD and reached three distinct peaks before returning to baseline concomitant with the interstitial DC potential of SD. The first peak occurred within 5 s of the triggering pulse, was smallest, and heralded the onset of SD. The second Ca2+ change was the greatest and reached a peak 6 s later, during the early phase of SD. The third was intermediate in size and occurred 18 s later, as SD reached its maximum interstitial DC change. SD was prevented by nonselective Ca2+ blockade (Ni2+ and Cd2+) but not by either L-Ca2+ channel (nifedipine) or N-Ca2+ channel inhibition (omega-conotoxin GVIA). Importantly, SD was blocked by P/Q Ca2+ channel antagonism (omega-agatoxin-IVA), which also prompted a significant reduction in pyramidal cell Ca2+ change and hyperexcitability. These results show that the spatiotemporal pattern of pyramidal cell Ca2+ change with SD is multiphasic; they provide further evidence that these changes begin before electrophysiologic evidence of SD. Furthermore, they show that P/Q Ca2+ channel antagonism can prevent SD in HOTCs and it appears to do so by preventing the NaAc induced increased pyramidal cell excitability from NaAc exposure, which may involve altered GABAergic transmission. PMID- 15132438 TI - Reduced basal CaMKII levels in hippocampal CA1 region: possible cause of stress induced impairment of LTP in chronically stressed rats. AB - Chronic psychosocial stress markedly reduces the expression of high-frequency stimulation (HFS)-evoked early long-term potentiation (LTP) in the CA1 region of the hippocampus of anesthetized rats. Immunoblotting was performed to determine changes in molecular levels of key signaling proteins that might be responsible for this inhibitory effect. Western blot analysis of the CA1 region demonstrates that chronic psychosocial stress decreases basal levels of calcium calmodulin kinase II (CaMKII), phosphorylated (P)-CaMKII, calmodulin, and protein kinase C (PKCgamma) while markedly increasing protein phosphatase 2B (calcineurin) levels. The decrease of basal levels of P-CaMKII may be triggered primarily by excessive dephosphorylation resulting from enhanced basal levels of calcineurin. The decline in the basal levels of the upstream molecules, PKCgamma and calmodulin may be a consequence of the diminished basal P-CaMKII levels. Analysis of signaling molecules in CA1 region of chronically stressed rat subjected to HFS in vivo showed only one difference compared to similarly stimulated control rats; no increase in P-CaMKII levels. Our results suggest that decreased P-CaMKII levels may be primarily responsible for the stress-induced reduction in LTP expression. PMID- 15132439 TI - Nickel content of as-received and retrieved NiTi and stainless steel archwires: assessing the nickel release hypothesis. AB - This study assesses the nickel content of as-received and retrieved stainless steel and NiTi archwires alloys. New and used brand-matched, composition-matched, and cross section-matched archwires were subjected to scanning electron microscopy and energy-dispersive electron probe microanalysis. Elemental analysis was performed on three randomly selected areas, and the nickel content, expressed as ratios of Ni/Ti (in NiTi wires) or Ni/Fe (in stainless steel), was statistically analyzed with a t-test (alpha = .05). No changes were detected with respect to Ni content ratios between as-received and retrieved NiTi or stainless steel wires, suggesting an absence of nickel release. Wear and delamination phenomena on the wire surface and the formation of galvanic couple between the stainless steel wires and bracket brazing materials intraorally may modify the corrosion susceptibility of the wire alloys in clinical conditions. PMID- 15132440 TI - The effects of differences in landmark identification on the cephalometric measurements in traditional versus digitized cephalometry. AB - The aim of this study was to explore the effects of differences in landmark identification on the values of cephalometric measurements on digitized cephalograms in comparison with those obtained from original radiographs. Ten cephalometric radiographs were randomly selected from orthodontic patients' records. Seven orthodontic residents identified 19 cephalometric landmarks on the original radiographs and digitized images. Twenty-seven cephalometric measurements were computed with a customized computer-aided program. To assess the concordance between cephalometric measurements derived from landmarks identified on the original radiographs and those from digitized counterparts, the values of 27 cephalometric measurements were compared to quantify the absolute value of measurement difference and the interobserver errors between these two methods. We found that the differences of all cephalometric measurements between original radiographs and their digitized counterparts were statistically significant. The differences in 21 of the 27 cephalometric items were less than two units of measurement (mm or degree), which is generally within one standard deviation of norm values in conventional cephalometric analysis. Moreover, statistically significant differences of interobserver errors between the two methods were noted only for seven of the 27 cephalometric items. In conclusion, the measurement differences between the original cephalograms and the digitized images are statistically significant but clinically acceptable. The interobserver errors for cephalometric measurements on our digitized cephalometric images are generally comparable with those on the original radiographs. The results of our study substantiated the benefits of digital cephalometry in terms of the reliability of cephalometric analysis. PMID- 15132441 TI - The esthetic properties of lips: a comparison of models and nonmodels. AB - It is perceived that fuller lips are more attractive, and hence lip augmentation has become common in esthetic plastic surgery. Numerous materials have been used, including collagen, autologous adipocytes, and more recently Restylane; however, little data exist on what comprises esthetically beautiful lips. Photographs of 28 models from fashion magazines were scanned to obtain digital images. These were selected strictly, using only exactly anterior-facing pictures. Using image analysis software, a range of defined lengths, angles, the lip area, and perimeter were measured. Lengths were expressed as a ratio of the intercanthal distance. A group of 14 nonmodel hospital employees were used as controls, with images obtained using a digital camera, and the same measurements were calculated. Results were compared for the two groups, and statistical analysis was performed using the Student's t-test. Overall lip width was not significantly different between the two groups (models, 15.7 units; nonmodels, 15.9 units). Both upper- and lower-lip height was significantly greater in models than in nonmodels (models, 2.1 and 3.6 units; nonmodels, 1.6 and 2.7 units, respectively), as was the upper-lip height laterally at the point of the angle of cupid's bow. Correspondingly, the angles of both upper and lower lips were also greater in models (models, 30.0 degrees and 47.3 degrees; nonmodels, 23.2 degrees and 37.6 degrees, respectively). We have assumed the model group to have esthetically beautiful lips. Our quantitative measurements have confirmed that this population has fuller lips compared with nonmodel controls, as determined by the lip height and angles. PMID- 15132442 TI - The influence of extraction treatment on Holdaway soft-tissue measurements. AB - The aims of this study were (1) to determine the changes in the soft-tissue measurements of orthodontic patients treated with four first premolars extractions as determined by the Holdaway Analysis, (2) to investigate the sexual differences between pre- and posttreatment values, and (3) to compare the Holdaway soft-tissue norms with findings in Anatolian Turkish adults. Pretreatment and posttreatment lateral cephalograms of 58 patients (26 boys and 32 girls) were evaluated. During treatment, the H angle, soft tissue subnasale to H line, and upper-lip strain decreased, and the upper-lip thickness, inferior sulcus to H line, and nose prominence increased. No statistically significant sex differences were found between the pre- and posttreatment values of the four premolar extraction cases. When the pretreatment values were compared with the Holdaway soft-tissue norms of Anatolian Turkish adults, seven variables showed statistically significant differences. When the posttreatment values were compared, six measurements showed statistically significant differences. It was determined that some measurements (H angle, inferior sulcus to H line, upper-lip thickness) moved closer to the Anatolian Turkish norms with extraction of four first premolars. However, upper-lip strain measurement changed in an undesirable direction during the treatment. In this study, the sample began treatment with greater facial imbalance, and the facial esthetics improved during treatment. It is necessary to investigate this difference in future studies using larger sample sizes. PMID- 15132443 TI - Effects of orthodontic treatment on mandibular rotation and displacement in Angle Class II division 1 malocclusions. AB - The aim of this retrospective cephalometric study was to investigate the effects of orthodontic treatment on rotation and displacement of the mandible in Angle Class II, division 1 malocclusions. Thirty patients in the treated group (15 boys and 15 girls; mean age at pretreatment, 12.27 +/- 1.36 years) were compared with 28 subjects who had untreated Class II, division 1 malocclusions (15 boys and 13 girls; mean age at T1, 12.01 +/- 0.07 years). The patients in the first group were treated nonsurgically, without extraction, and without the use of functional appliances. Cephalometric data were obtained from three lateral cephalograms per case representing pretreatment (T1), posttreatment (T2), and at least 2-years postretention (T3). Thirty-seven variables were measured representing craniofacial morphology, tooth measurements, and mandibular displacement. Some variables were obtained from cranial base, maxillary, or mandibular superimposition. Statistical significance was established at P < .05, P < .01, and P < .001. The findings indicated that orthodontic treatment of Class II, division 1 malocclusions induced a more vertical mandibular growth direction associated with an increased vertical displacement of pogonion. Occlusal or vertical movement of maxillary and mandibular molars was not correlated to mandibular rotation or horizontal displacement of pogonion. When compared with controls, the treated group did not exhibit a significant difference in mandibular rotation or occlusal movement of maxillary molars; however, it did show a greater occlusal movement of mandibular molars during treatment. PMID- 15132444 TI - A comparison of the effects of rapid maxillary expansion and fan-type rapid maxillary expansion on dentofacial structures. AB - The aim of this study was to evaluate and compare the sagittal, transverse, and vertical effects of rapid maxillary expansion (RME) and fan-type RME on dentofacial structures. The study group consisted of 34 patients, 14 boys and 20 girls (average age 12.5 years), selected without considering their skeletal class and sex. The fan-type RME group comprised 17 subjects, who had an anterior constricted maxilla with a normal intermolar width. The RME group comprised 17 other subjects, who had a maxillary transverse discrepancy with a posterior crossbite. The records obtained for each patient included a lateral and a frontal cephalometric film, upper plaster models, and occlusal radiograph obtained before treatment (T1), after expansion (T2), and immediately after a three-month retention period (T3). The data obtained from the evaluation of the records before and after treatment, after treatment and after retention, and before treatment and after retention were compared using paired t-test. Further comparisons between the groups were made using Student's t-test. There was significantly greater expansion in the intercanine than in the intermolar width in the fan-type RME group as compared with the RME group. Downward and forward movement of the maxilla was observed in both groups. The upper incisors were tipped palatally in the RME group, but they were tipped labially in the fan-type RME group. There was significantly greater expansion in the nasal cavity and maxillary width in the RME group as opposed to the fan-type RME group. PMID- 15132445 TI - Vertical forces in labial and lingual orthodontics applied on maxillary incisors- a theoretical approach. AB - Theoretical and experimental biomechanical analyses explain most labial orthodontics (LaO); however, lingual orthodontic (LiO) biomechanical principles are rarely introduced. The objective of this study was to apply basic biomechanical considerations in understanding the influence of maxillary incisor inclination and to compare the effect of labial vs lingual intrusive/extrusive forces on tooth movement. Basic anatomic and geometric hypotheses were assumed, ie, tooth length (crown and root), location of the center of resistance, and crown thickness. Incisor inclination as related to a perpendicular line to the occlusal plane (OP) varied between -35 degrees (retroclination) and 45 degrees (proclination). A 0 degrees inclination was defined as a tooth position with its long axis perpendicular to the OP. The buccolingual moment for characterizing root movement was calculated for an applied force perpendicular to the OP. The results showed that when using LaO, an extrusion force resulted in labial root movement from a retroclination of 20 degrees up to a proclination of 45 degrees. In LiO, labial root movement occurred only when the tooth was proclined more than 20 degrees. In all other tooth inclinations, lingual root movement occurred. The opposite tooth movement occurred when an intrusive force was applied. Application of a vertical force has different clinical effects on tooth movement with labial and lingual appliances. Application of a lingual force is more complicated, and its effect on tooth movement depends on bracket position and initial tooth inclination. PMID- 15132446 TI - Evaluation of the frictional resistance of conventional and self-ligating bracket designs using standardized archwires and dental typodonts. AB - The frictional behavior of four conventional and four self-ligating brackets were simulated using a mechanical testing machine. Analyses of the two-bracket types were completed by drawing samples of three standardized archwires through quadrants of typodont models in the dry/wet states. Pretreatment typodonts of an oral cavity featured progressively malocclused quadrants. As nominal dimensions of the archwires were increased, the drawing forces of all brackets increased at different rates. When coupled with a small wire, the self-ligating brackets performed better than the conventional brackets. For the 0.014-inch wires in the upper right quadrant, the maximum drawing forces averaged 125 and 810 cN for self ligating and conventional brackets, respectively. When coupled with larger wires, various designs interchangeably displayed superior performance. For the 0.019- x 0.025-inch wires in the upper left quadrant, the maximum drawing forces averaged 1635 and 2080 cN for self-ligating and conventional brackets, respectively. As the malocclusion increased, the drawing forces increased. For example, in the least malocclused quadrant and with the smallest wire, maximum drawing forces for self-ligating and conventional brackets averaged 80 and 810 cN, respectively, whereas in the most malocclused quadrant tested with the same wire size, maximum drawing forces for self-ligating and conventional brackets averaged 870 and 1345 cN, respectively. For maximum values between the dry and wet states, significant differences between ambient states existed only for the In-Ovation brackets in the lower left quadrant. These test outcomes illustrated how bracket design, wire size, malocclusion, and ambient state influenced drawing forces. PMID- 15132448 TI - Statistical evaluation of possible factors affecting the sagittal position of the first permanent molar in the maxilla. AB - A cephalometric analysis was designed to evaluate several factors that may affect the sagittal position of the maxillary first permanent molar. A total of 184 Class II and Class I malocclusion patients were randomly selected before orthodontic treatment. The mandibular and palatal planes were related to Frankfort Horizontal and used to create the interjaw or B angle. Age and cephalometric landmarks (Ba, N, point A, pterygomaxillary fissure, and maxillary molars) were projected at right angles to the Frankfort Horizontal for effective length. Actual maxillary length and actual molar location were determined by projecting landmarks at right angles to the palatal plane. Correlation coefficients and P values were used to evaluate the data with a minimal significance value of .05 to determine a 95% confidence level. A statistically significant linear and proportional positive correlation (P < .0001) existed between molar location, age, and maxillary size. There was a strong negative correlation (P < .0001), both linearly and as a proportion of the actual length of the maxilla, between the actual position of the maxillary molar and the interjaw and mandibular plane angles. A significant correlation also existed between the molar position and palatal plane angles. The results show that increased interjaw, mandibular, and palatal plane angles are accompanied by a more posterior position of the maxillary first molar in the maxilla, whereas the molar occupied a continuing more forward position in the maxilla with increasing age, cranial base length, and maxillary size. PMID- 15132447 TI - Comparison of skeletal and dental morphology in asymptomatic volunteers and symptomatic patients with unilateral disk displacement with reduction. AB - The purpose of this study was to evaluate the effect of unilateral disk displacement with reduction (UDDR) on the skeletal and dental pattern of affected individuals. There were 18 symptomatic female patients and 46 asymptomatic normal female volunteers. All study participants had bilateral high-resolution magnetic resonance scans in the sagittal (closed and open) and coronal (closed) planes to evaluate the temporomandibular joints. Linear and angular cephalometric measurements were taken to evaluate the skeletal, denture base, and dental characteristics of the two groups. Analysis of variance was used to compare the symptomatic with the control subjects. A few skeletal differences were found. There was an overall reduction in length of the anterior (S-Na) and posterior (S Ba) cranial base measurements in the UDDR group. The cranial base angle was also increased. Both upper and lower dentures bases were retropositioned. The posterior ramal height (Ar-Go) was shorter in the symptomatic group. This study showed that alterations in skeletal morphology may be associated with UDDR. The mechanisms that produce DD or the mechanisms that cause this skeletal alteration are yet to be clarified. This study suggests that subjects with UDDR may manifest altered craniofacial morphology. The clinician should be aware of this possibility, especially for growing patients. PMID- 15132449 TI - Comparison of the effect on oral discomfort of two positioning techniques with lingual brackets. AB - Using a standardized questionnaire, 41 patients (12 men, 29 women; mean age 31.5 +/- 12.1 years) were interviewed prospectively on their subjective oral comfort, various oral functions, and professional qualification before indirect application of lingual brackets (T0), within 24 h of application (T1), and three months later (+/- 1 week) (T2). In 22 of the 41 patients, the brackets were positioned with the Bonding with Equalized Specific Thickness (BEST) technique (BEST group) and in 19 patients by the Transfer Optimized Positioning (TOP) method (TOP group). Despite positive adaptation, the patients in both groups still reported a significant deficiency in tongue space at T2 as well as significantly more frequent lesions to the tongue. The BEST group was affected significantly more often by these problems than the TOP group. At T2, the tongue position was also rated as changed significantly more often in the BEST group than in the TOP group. Although the BEST positioning technique leads to greater impairments in oral comfort than the TOP technique, it offers the orthodontist the advantage of less bending input. In both techniques, there is a need for detailed briefing of patients about the extent and duration of impairments induced by lingual brackets. PMID- 15132450 TI - An in vitro study simulating effects of daily diet and patient elastic band change compliance on orthodontic latex elastics. AB - This project investigated the effects of food exposure and patient compliance with elastic-band change on the degradation of forces in 3/16-inch, medium-wall, latex elastic bands during a simulated day of clinical wear. Six levels of daily diet/patient compliance were chosen as representative of orthodontic patients and a quasicontrol group. The groups differed with respect to how much exposure to artificial saliva and foodstuffs they experienced. After exposure in mild tension to daily diets and based on compliance with instructions about changing orthodontic elastics, the elastics were tested in tensile mode by stretching to 25 mm, where the load was recorded in newtons. The bands of three manufacturers, Rocky Mountain Orthodontics (RMO), 3M Unitek (UNO), and American Orthodontics (AMO), were examined, with 10 bands per group, per manufacturer, forming a cohort. Two-way analysis of variance and the Tukey-Kramer honestly significant difference tests were used to identify statistical significance (P > .05). With respect to bands from a single manufacturer, no differences were found between daily diet/patient compliance levels. However, differences (P < .0001) were found between manufacturers' bands. RMO > UNO > AMO in all environments. Over a 24-hour period, latex elastics maintain their applied load in the simulated oral environments. PMID- 15132451 TI - Evaluation of the dental plaque pH recovery effect of a xylitol lozenge on patients with fixed orthodontic appliances. AB - The purpose of this study was to evaluate the influence of a xylitol lozenge on the dental plaque pH profile of fixed orthodontic patients. Twelve volunteers participated in this study. Before the measurement of plaque pH, subjects were asked to refrain from brushing their teeth for 48 hours and from eating and drinking for two hours. The subjects' baseline dental plaque pH was recorded using the touch technique. It was followed by a one-minute rinse with 15 ml of a 10% solution of sucrose, and subsequent plaque pH measurements were carried out during the next one hour. Xylitol lozenges were taken five times a day during a 14-day period. The variables of resting-plaque pH, minimum-plaque pH (MP pH), time required to reach MP pH (TMP), last-plaque (LP) pH at the end of one hour, cH area (CH), and pH at each test time were calculated for each pH test of the subjects. The paired sample t-test was used for statistical comparison. The mean MP pH values increased from 4.81 to 5.09 in the experimental measurement (P < .05). The mean TMP was not affected by the use of xylitol (P > .05). Although the LP pH showed an increase during the experimental period, the difference between control and experimental periods was not statistically significant (P > .05). The CH of the experimental period was significantly less than that of the control period (P < .05). As a result, the use of a xylitol lozenge after a sucrose challenge can be an advisable practice for fixed orthodontic patients to prevent future dental caries. PMID- 15132452 TI - Effect of time on bond strength in indirect bonding. AB - The purpose of this in vitro investigation was to determine the influence of a reduced time interval before debonding on shear bond strength of stainless steel brackets bonded with a custom base indirect technique. A total of 135 bovine permanent mandibular incisors was randomly divided into nine groups of 15 specimens each. Three base composite-sealant combinations were investigated: (1) Phase II base composite, Custom I.Q. sealant, (2) Phase II base composite, Maximum Cure sealant, and (3) Transbond XT base composite, Sondhi Rapid Set sealant. Shear bond strength was measured for three different debonding time intervals: (1) time of transfer tray removal as recommended by the manufacturer, (2) 30 minutes after bonding of the sealant, and (3) 24 hours after bonding of the sealant. For groups bonded with Maximum Cure or Sondhi Rapid Set sealants, no influence of debonding time on shear bond strength was found. The Custom I.Q. sealant groups showed significantly lower bond strength measurements when debonded at the recommended tray removal time, and the Weibull analysis indicated a higher risk of bond failure at clinically relevant levels of stress. All base composite-sealant combinations showed acceptable bond strength at 30 minutes and 24 hours after bonding of the sealant. PMID- 15132453 TI - Efficacy of using self-etching primer with a 4-META/MMA-TBB resin cement in bonding orthodontic brackets to human enamel and effect of saliva contamination on shear bond strength. AB - The objective of this study was (1) to evaluate the effectiveness of Megabond when used with Superbond C&B, a 4-methacryloloxyethyl trimellitate anhydride (4 META)/methyl methacrylate (MMA)-tri-n-butyl borane (TBB) resin, to bond orthodontic metal brackets to human enamel and (2) to examine the influence of saliva contamination on shear bond strength. Metal brackets were bonded to phosphoric acid-etched or Megabond-treated human premolars using Superbond C&B resin cement. The effects of saliva contamination after acid etching or self-etch priming, and the effect of re-etching or self-etch priming after saliva contamination on shear bond strength were also assessed. The shear bond strengths were measured after immersion in water at 37 degrees C for 24 hours. Data were analyzed using two-way analysis of variance and Fisher's protected least significant difference test for multiple comparisons. There were no significant differences in shear bond strength between phosphoric acid etching and self-etch priming for no contamination, saliva contamination, and repeat treatment (etching or priming) after saliva contamination. With phosphoric acid etching, saliva contamination significantly decreased the shear bond strength. Repeat phosphoric acid etching after saliva contamination did not significantly improve the bond strengths. With self-etching primer treatment, however, saliva contamination did not cause any decrease of bond strength. Phosphoric acid etching produced more enamel fracture than self-etching primer treatment. Field-emission scanning microscopy revealed less dissolution of enamel surface resulted from self-etching primer compared with phosphoric acid. These results suggest that Megabond when used with Superbond C&B resin cement may be a good candidate for bonding orthodontic brackets to human enamel. PMID- 15132454 TI - Effect of light-emitting diode on bond strength of orthodontic brackets. AB - The aim of this study was to evaluate the effect of light-emitting diode (LED) light curing on shear bond strength (SBS) of orthodontic brackets bonded to teeth. Light exposure of 40 seconds from a conventional halogen-based light curing unit was used as a control. Eighty human premolars were divided into four groups of 20 each. Brackets were bonded to acid-etched teeth with Transbond XT light-cured adhesive. In the first group, the adhesive was light cured for 40 seconds with a conventional halogen unit (XL3000, 3M). In the other three groups, adhesive was cured with a commercial LED unit (Elipar FreeLight, 3M ESPE) for 10, 20, or 40 seconds. SBS of brackets was measured on a universal testing machine and recorded in megapascals. Adhesive remnant index (ARI) scores were determined after failure of brackets. Data were analyzed using analysis of variance and chi square tests. No statistically significant differences were found among the SBS values of halogen-based light-cured (13.1 +/- 3.1 MPa) and 20- and 40-second LED cured (13.9 +/- 4.8 MPa and 12.7 +/- 5.1 MPa) specimens (P > .05). However, 10 seconds of LED curing yielded significantly lower SBS (P < .05). No statistically significant differences were found between the ARI scores among groups. The results of this study are promising for the orthodontic application of LED-curing units, but further compatibility and physical characteristic studies of various orthodontic adhesives and clinical trials should be performed before validation. PMID- 15132455 TI - A comparison of shear bond strength and debonding characteristics of conventional, moisture-insensitive, and self-etching primers in vitro. AB - Bond failure is often attributed to moisture contamination. To overcome this commonly encountered problem, materials have been developed that are hydrophilic and that are believed to offer better bond strength in moisture-contaminated environment. Shear bond strength was compared among three materials: conventional Transbond XT primer (3M Unitek), moisture-insensitive primer (MIP, 3M Unitek), and self-etch primer (Transbond plus, 3M Unitek). Bond strength was tested under laboratory conditions with brackets bonded on both dry enamel and enamel contaminated with natural saliva. Self-etch primer showed maximum bond strength under both dry and wet conditions. Conventional primer was comparable with the former under dry conditions but did not offer clinically adequate bond strength in cases of moisture contamination. Both MIP and self-etch primer showed adequate bond strength superior to that of conventional primer in case of moisture contamination. All primers showed typical debonding characteristics of separation at the bracket-adhesive interface or within the adhesive itself, with the exception of the conventional primer used with moisture-contaminated enamel. PMID- 15132456 TI - A systematic review of the consequences of premature birth on palatal morphology, dental occlusion, tooth-crown dimensions, and tooth maturity and eruption. AB - This systematic review addresses the question whether prematurity results in alteration of palatal morphology, dental occlusion, tooth-crown dimensions, and tooth maturation. A literature survey from the PubMed database covering the period from January 1966 to November 2002 used the Medical Subject Headings terms "infant, premature," and "infant, low birth weight" in combination with "jaws," "dental physiology," "dentition," and "tooth abnormalities." Controlled studies written in English and with definitions of premature birth according to the World Health Organization were selected. Two reviewers selected and extracted the data independently and also assessed the quality of the studies. The search strategy resulted in 113 articles, of which 13 met the inclusion criteria. Scientific evidence was found for altered palatal morphology in the short term among the premature children, and oral intubation was a contributing factor to the alterations. If corrected age was considered for the premature children, no delay in dental development and eruption was found compared with normally born children. Thus, the early birth of premature children must be taken in account when planning for orthodontic treatment. Because of the contradictory results and lack of longitudinal studies, the scientific evidence was too weak to answer the questions whether premature birth causes permanent alteration of palatal morphology, alteration of dental occlusion, and altered tooth-crown dimensions. To answer these questions and obtain reliable scientific evidence whether premature children are at risk for malocclusions from possible alterations of palatal morphology such as asymmetry and high arched palates, further well designed controlled studies as well as longitudinal studies are needed. PMID- 15132457 TI - Severe Class II anterior deep bite malocclusion treated with a C-lingual retractor. AB - A C-lingual retractor was placed on the lingual aspects of the six maxillary anterior teeth in a 24-year-old female patient with a Class II anterior deep-bite malocclusion. The treatment plan consisted of extracting both the upper first premolars and intruding and retracting the upper six anterior teeth. Transpalatal arches were soldered to the upper first and second molar bands and used as an intra-arch anchor unit for upper space closure. Double NiTi closed coil springs were used palatally between the hooks of the C-lingual retractor and the transplantar arches. A high-pull headgear was used for anchorage reinforcement during en masse retraction. It took 14 months to treat this patient. The correct overbite and overjet was obtained by simultaneously intruding and retracting the upper six anterior teeth into their proper positions by C-lingual retractor mechanics, which contributed to an improvement in facial balance. The treatment result was stable 6 months after debonding. The application of this new appliance, consideration in case selection, and sequence of treatment are presented. PMID- 15132458 TI - Generations of training. PMID- 15132459 TI - Preoperative chemoradiation for rectal cancer: kudos and a caution. PMID- 15132460 TI - Snowmobile trauma: 10 years' experience at Manitoba's tertiary trauma centre. AB - INTRODUCTION: According to the literature, the increased recreational use of the snowmobile has resulted in an increasing number of musculoskeletal injuries. We wished to examine whether previously described risk factors continue to be associated with snowmobile trauma and to identify previously unrecognized risks and specific patterns of injury. METHODS: We carried out a chart review of all snowmobile-related injuries over a 10-year period at the Health Sciences Centre in Winnipeg, the only level 1 trauma centre serving the Province of Manitoba, with particular attention to the risk factors of suboptimal lighting, excessive speed and alcohol consumption. RESULTS: We identified 480 injuries in 294 patients, and 81 (27.6%) of these patients died. Collisions accounted for 72% of the injury mechanisms. Of the injuries sustained, 31% occurred on roads. Excessive speed was a risk factor in 54% of patients, suboptimal lighting in 86% and a blood alcohol level greater than 0.08 in 70%. Musculoskeletal injuries accounted for 57% of those recorded. There were also brachial plexus injuries (3%) and knee dislocations (2%). To our knowledge, this is the largest study detailing injury associated with recreational use of snowmobiles in Canada. CONCLUSIONS: Because snowmobile trauma is caused principally by human errors, it is potentially preventable. Efforts aimed at prevention must focus on the driver, who controls the common risk factors. The danger of snowmobiling while intoxicated must be emphasized. Trail-side monitoring is likely to be ineffective, as the majority of accidents do not occur on designated snowmobile trails. PMID- 15132461 TI - Extrapulmonary intrathoracic hydatid cysts. AB - INTRODUCTION: Hydatid disease, a clinical entity endemic in many sheep- and cattle-raising areas, is still an important health problem in the world. Extrapulmonary location of cysts in the thorax is rare. We report our experience with intrathoracic but extrapulmonary hydatid cysts and discuss concepts of treatment. METHOD: In our Thoracic and Cardiovascular Surgery Department at the Dicle University School of Medicine, 133 patients with thoracic hydatid cysts were managed surgically between January 1990 and October 2002. In 14 (10.5%), the cysts were extrapulmonary but within the thorax, located in the pleural cavity, mediastinum, pericardium and diaphragm, or in pleural fissures. Cysts were intact in 12 patients and ruptured in 2. Radiographs of the chest were the main means of diagnosis; all patients with mediastinal and diaphragmatic cysts and some with pleural cysts were also scanned with computed tomography. All patients were managed surgically. RESULTS: We operated on 3 mediastinal, 2 diapragmatic and 1 pericardial hydatid cyst, as well as 6 in pleural fissures and 2 in the pleural space. Lateral thoracotomy was chosen as the surgical incision in all patients except 1 (7% of the 14), who had median sternotomy for a pericardial hydatid cyst. Empyema developed in 2 patients (morbidity, 14%). No patient died perioperatively. CONCLUSIONS: Hydatid cysts may be found in many different sites. Surgery to obtain a complete cure is the treatment of choice for most patients with intrathoracic but extrapulmonary cysts; excision must be done without delay to avoid or relieve compression of surrounding vital structures. PMID- 15132462 TI - Hemipelvectomy: a changing perspective for a rare procedure. AB - OBJECTIVE: To compare the prognosis of patients undergoing a hemipelvectomy (HP) in the treatment of pelvic sarcomas and carcinomas and to review the morbidity and mortality associated with HP. DESIGN: Retrospective chart review. SETTING: The Foothills Hospital, University of Calgary, Calgary, Alberta. PATIENTS: Thirteen patients with clinically and radiographically isolated malignancies involving the bony pelvis and adjacent structures. INTERVENTIONS: Patients were treated with either an external HP (9 patients) or internal HP (4) in 1983-2001. OUTCOME MEASURES: Survival and recurrence rates for patients in 2 histopathologic groups (sarcoma v. carcinoma); morbidity and mortality associated with HP. RESULTS: Hemipelvectomy was performed for 7 sarcomas (4 primary bone and 3 soft tissue) and 6 carcinomas (5 genital tract and 1 unknown primary). Seven of the 9 external HPs involved composite resection of other pelvic structures, including other pelvic viscera (3 patients), sacrum (3) and portions of lumbar vertebrae and nerves (1). There were no additional resections among the 4 internal HPs, but 3 patients had allograft reconstruction. Length of stay averaged 30 days (range 14-70 d). At least 1 complication occurred in 10 of 13 cases. The most common complication was flap necrosis occurring in 5 patients (38%). There was 1 perioperative death (8%). The survival of patients treated for sarcomas was better than for carcinomas, which were primarily of the genital tract. Only 1 of the patients with a pelvic sarcoma died of disease (86% disease-specific survival), with a median follow-up of 12 months (range 9-108 mo). Of the 7 sarcoma patients 5 were disease-free at last follow-up. One of 6 pelvic carcinoma patients died perioperatively, with another dying of unknown causes 4 months after surgery. Of the 4 remaining patients 3 died of disease, resulting in a median survival of 9 months (range 4-20 mo). Four of 6 patients with pelvic carcinomas developed recurrent disease, none local. CONCLUSIONS: HP has considerable morbidity but is a viable and potentially curative treatment for patients with pelvic sarcomas. With pelvic carcinomas HP was not curative, but did provide short-term local disease control. Future improvements in imaging techniques and quality-of-life studies may help with patient selection. The role of HP in recurrent carcinoma remains to be determined. PMID- 15132463 TI - Current awareness in Canada of clinical practice guidelines for colorectal cancer screening. AB - INTRODUCTION: The Canadian Task Force on Preventive Health Care (CTF-PHC) recently revised its screening recommendations for colorectal cancer (CRC). We wished to assess the effect of this change on the screening beliefs and clinical practice of primary care physicians. METHODS: We surveyed 160 primary-care physicians, quasi-randomly sampled, in June-July 2001 and again in April-July 2002, 9 months after publication of the guidelines. Descriptive statistics and McNemar chi2 analyses were carried out on data from physicians who responded to both surveys. RESULTS: Of the those sampled, 47% responded to both surveys. After the publication of the CTF-PHC guidelines, the proportion reporting that they recommend CRC screening to their patients at average risk increased from 43% to 60% (p = 0.02). Before publication of the revised guidelines 48% stated that the CTF-PHC did not support screening, compared with 24% afterward (p = 0.01). CTF PHC guidelines were acknowledged by 30% to be a source of CRC screening information. Around 9 months post-publication, 24% of the physicians stated their awareness of the revised screening guidelines. The most commonly cited reasons for not recommending CRC screening to average-risk patients were that the evidence is inconclusive and that CTF-PHC guidelines do not support screening. CONCLUSIONS: After publication of the revised CTF-PHC guidelines more primary care physicians reported that they recommend CRC screening to their average-risk patients. The belief that the evidence is inconclusive nevertheless remains a considerable barrier to implementation. To increase the use of screening for CRC, additional strategies are required. PMID- 15132464 TI - Enteral nutrition and mucosal immunity: implications for feeding strategies in surgery and trauma. AB - Systemic inflammatory responses to severe trauma and surgical illnesses may be partly responsible for numerous complications, including sepsis, multiple organ failure and unregulated hypermetabolism leading to protein-calorie malnutrition. The integrity of the gastrointestinal tract appears to be an important factor in the pathogenesis of the systemic inflammatory response and sepsis. Resuscitation and nutrition support strategies for preserving gut mucosal integrity have therefore been strongly promoted. This review summarizes the scientific rationale for emphasizing enteral nutritional support of surgical patients, discusses some important limitations of enteral feeding and argues for a flexible approach to nutrition support for these complex patients. PMID- 15132465 TI - Surgical images: musculoskeletal. Delayed cauda equina syndrome due to a sacral insufficiency fracture missed after a minor trauma. PMID- 15132466 TI - Radiology for the surgeon: soft-tissue case 54. Spigelian hernia [correction]. PMID- 15132467 TI - Surgical Education and Self-assessment Program (SESAP). Recurrent pyogenic cholangitis. PMID- 15132468 TI - Transplantation: focus on kidney, liver and islet cells. AB - Over the past decade, advances in immunosuppression, organ preservation, surgical techniques and perioperative management have resulted in improved survival rates for solid organ transplants. Even so, the field of transplantation still presents many challenges. A critical obstacle is the shortage of donor organs. The paucity of cadaveric organs has increased the demand for living donor transplantation. Although this option has expanded the organ pool, concerns over ethical issues and donor safety remain, and there is an ongoing effort to make living donation a safer and less invasive process. An alternative to solid organ transplantation involves the transplantation of cells, such as islet cells for type 1 diabetes mellitus. Whereas transplantation of solid organs has seen steady improvement over the past 2 decades, transplantation of islet cells has not. Recent advances in the field of islet cell transplantation, however, have made this procedure a clinical reality. Stem cell research has provided a glimpse into the possible future of transplantation for organ failure. Another major barrier to transplantation is the lifelong need for immunosuppression. Current immunosuppression protocols place transplant recipients at continuing risk for immunosuppression-associated complications such as infection and malignant disease. New agents continue to reduce the rates of acute graft rejection and to increase long-term survival; however, they have exposed metabolic and cardiovascular complications without affecting the incidence of chronic rejection. The ultimate goal of many investigators in this field is to achieve specific immunologic graft tolerance. In this article we summarize recent technical advances in the field of transplantation that address some of the challenges. PMID- 15132469 TI - Total mesorectal excision: technical aspects. AB - Total mesorectal excision (TME) is a precise dissection of the rectum and all pararectal lymph nodes within an oncologic package: the mesorectal envelope. This article is a brief description of the technical aspects of the dissection, illustrated by cadaver dissection. Here the TME dissection is organized into 6 steps to facilitate learning: (1) left retroperitoneum; (2) superior rectal and inferior mesenteric vessels; (3) upper mesorectum; (4/5) right and left mid mesorectum; and (6) distal mesorectum and anorectal junction. The relationship of the autonomic nerves, blood vessels and adjacent organ structures are described at each step. The 6 steps are recommended for learning and performing TME dissection in cadavers and patients. PMID- 15132470 TI - Congenital diastasis of the inferior tibiofibular joint: case report and treatment analysis. PMID- 15132471 TI - Management of rectal lymphoma. PMID- 15132472 TI - Elevated parathyroid hormone-related peptide in a patient with an extragonadal germ-cell tumour and hypercalcemia. PMID- 15132473 TI - Total knee replacement surgery and surgical site infection: a prospective audit. PMID- 15132474 TI - Diverticulitis of the appendix. PMID- 15132475 TI - Early presentation of splenic injury after colonoscopy. PMID- 15132476 TI - Accreditation of residents. PMID- 15132477 TI - Tibial plateau fractures. PMID- 15132479 TI - Outreach multi-beneficial. PMID- 15132478 TI - SESAP question inaccurate. PMID- 15132480 TI - The absolutely fabulous but flawlessly customary world of female impersonators. AB - Our editorial introduction to this volume on drag queens highlights what we believe are some of the most prominent and important themes of female impersonation in the past and today. Building on contributors' articles, a substantial body of literature on female impersonators/drag queens and the social construction of gender, and our own extensive ethnographic experiences in a multitude of drag settings, we first suggest that such individuals can be seen as symbolic representatives of the cultural ideals associated with the feminine and women and how they have changed over time. We next argue that the notion of the effeminate drag queen is more a myth than a reality with the contextual benefits many performers receive-status and power-being indicative of the hegemony of masculinity in male-dominated societies. We next explore how additional social identities, such as race, class, nation, and religion, often impact drag performances and how others interpret them. We end our introduction by offering a model that delineates what are some of the present transgressive limits and subversive possibilities of female impersonation. PMID- 15132481 TI - A lovely war: male to female cross-dressing and Canadian military entertainment in World War II. AB - This article explores the Canadian military entertainment units during World War II (WWII), specifically those formed by the Navy, Army and Air Force from talent found amongst their own personnel. These entertainment units toured extensively in Canada, the United Kingdom and Europe with the goal of increasing the morale of combat troops while encouraging the enlistment of Canada's domestic populations in the war effort generally and the armed forces specifically. By focusing on male to female cross-dressing in the performances of these entertainment units and their pre-WWII antecedents, it will become clear that the nature and importance of the representation of femininity within the virtually all-male milieu that existed near the battlefront changed over time in response to the demands of the audiences. Until the second half of WWII, soldier audiences were generally unwilling to form any ideological links between cross-dressing and homosexuality. Female impersonators were the key cast members in troop shows during the Great War, but eventually fell out of favor in the last years of WWII after women were recruited in large numbers into the Canadian military and thus its entertainment infrastructure. With women then on the military stage, men who persisted in female impersonation were decreasingly popular with audiences, ultimately under growing suspicion of being homosexuals and gradually removed from the productions. PMID- 15132482 TI - Wigs, laughter, and subversion: Charles Busch and strategies of drag performance. AB - This paper examines the strategies of drag performer/ playwright Charles Busch. His performance aesthetic is explored and shown to be subversive even though its initial impulse is to entertain. Basing my arguments on the work of Judith Butler, Elin Diamond, and others, I argue that drag queens like Busch can not only entertain but also make audiences question and criticize through drag's power to create a Brechtian alienation effect and historicize the subject. After showing how he can be viewed as a drag queen, I give a brief biography and discuss such contested terms as "camp" and "gay sensibility." I then focus on Busch's staged reading of Ibsen's Hedda Gahler andA Doll's House, both done in one afternoon at Theatre for The New City (6 May 2000). By examining the performance of Busch and his fellow actors, I demonstrate how a contemporary relevancy is achieved by having the roles played by a female impersonator whose acting choices are filtered through a gay sensibility. The ongoing dialectic between spectator and performer creates a historicized moment in performance that underscores the gender dynamics in unexpected and stimulating ways. PMID- 15132483 TI - The beauty and the beast: reflections about the socio-historical and subcultural context of drag queens and "Tunten" in Berlin. AB - In this article, I focus on two different faces found in Berlin's gay subculture: the Tunten and the drag queens. Both are commonly seen as "male homosexual transvestites," although many such individuals today prefer to identify themselves somewhere within a diverse transgender spectrum rather than as transvestites. Tunten and drag queens differ in their gender performativity, their self-image and their chosen role models as well as in the niches in which they have been able to establish themselves in German mainstream society. Based on ethnographic data, I argue against the widespread reductionist view that the differences between Tunten and drag queens lie primarily in style, behavior, talent and success. Nor can these differences be easily explained away as a result of subculture globalization. Instead, I show that there is a simultaneous coexistence of both a subculturally established, "traditional" local transgender culture and a more recently adopted and partly imported, new local transgender culture. The coexistence of these two urban transgender cultures also indicates the paradigm shift in German gay and youth cultures of the last decades. Thus, I will emphasize the importance of the socio-historical and subcultural processes in studying transgender cultures in Western societies. PMID- 15132484 TI - Moffies, artists, and queens: race and the production of South African gay male drag. AB - This article draws on seventeen months of ethnographic fieldwork in South Africa to explore the experiences of urban and township drag performers. I show that two distinct sex-gender-sexuality systems have emerged based in the sociopolitical history of South Africa, and I argue that urban drag produces race oppositionally and examine how township femininity creates raced forms of gender, sex, and sexuality. Contemporary South African drag foregrounds the performativity and constitution of race and gender. My analysis attempts to challenge definitions of "drag" and "audience," suggesting the necessity for an integrated reconceptualization of drag studies. PMID- 15132485 TI - Ad/dressing the nation: drag and authenticity in post-apartheid South Africa. AB - This paper examines a style of drag in South Africa that features "traditional African" clothing. In a region in which homosexuality is denigrated as a colonial, European import and "unAfrican," the meaning of "traditional drag" is deeply inflected by the question of cultural authenticity. This dragging practice fits within a distinctly post-colonial production of tradition and its self conscious display--in the form of attire--of a decidedly "gay" one. Traditional drag also responds to ongoing politics within and between lesbian and gay communities about racial "representivity" and "transformation." The paper focuses on displays of traditional drag at Johannesburg's Gay and Lesbian Pride Parade but also explores the complex politics of publicity and address suggested by varying contexts in which traditional dress and drag are mobilized. PMID- 15132486 TI - Chicks with dicks, men in dresses: what it means to be a drag queen. AB - One of the burning questions about drag queens among both scholars and audiences is whether they are more gender-revolutionaries than gender-conservatives. Do they primarily destabilize gender and sexual categories by making visible the social basis of femininity and masculinity, heterosexuality and homosexuality? Or are they more apt to reinforce the dominant binary and hierarchical gender and sexual systems by appropriating gender displays and expressing sexual desires associated with traditional femininity and institutionalized heterosexuality? We address this question through a case study of drag queens at the 801 Cabaret in Key West, Florida. On the basis of life histories, observations of their performances, and focus groups with audience members, we examine the role of gender and sexuality in the process of becoming a drag queen and in the personal identities of drag queens. We find that transgenderism, same-sex sexuality, and theatrical performance are central to the personal identities of these drag queens, who use drag to forge personal and collective identities that are neither masculine nor feminine, but rather their own complex genders. PMID- 15132487 TI - "Let the drag race begin": the rewards of becoming a queen. AB - Drawing upon my ethnographic experiences in a drag venue called The Park in Roanoke, Virginia, this article explores the experiences of female impersonators in terms of their early motivations for doing drag, how they create and maintain drag personas and identities, and the obstacles to becoming a queen. Departing from previous researchers that have framed female impersonation as a deviant, stigmatizing, and pathological activity, this research analyzes the significant benefits some drag queens garner by donning women's attire. An experiential understanding of drag reveals that the significant rewards from the activity- contextual power and status, self-affirmation and empowerment--are powerful motivating factors. Instead of being deviant and/or partaking in pathological behavior, female impersonators can be seen as operating on an incentive system where the benefits of doing drag positively enrich the quality of the performer's life in a context where successful queens are held in the highest regard. PMID- 15132488 TI - Transformance: reading the gospel in drag. AB - Despite the large body of scholarship on drag and its performance of misogyny, mimicry, and masculinity, little attention has been paid to the role of musical genres in Black drag performance and its reception. This essay explores drag performances of gospel music and its relationship with the spectator at the Biology Bar, a Black gay drag site in Chicago. By examining the shift from the club "space" to the church "place," this research locates several possibilities for queer gospel performances. Through the introduction of a theory of transformance, this essay highlights the contradictions, complications, and complexities of the relationship between the Black church and the Black gay community. PMID- 15132489 TI - Kind of a drag: gender, race, and ambivalence in The Birdcage and To Wong Foo, Thanks for Everything! Julie Newmar. AB - This paper examines the ways in which two Hollywood films featuring drag queens, Too Wong Foo, Thanks for Everything! Julie Newmar and The Birdcage, offer a kind of "both/and" look at the complexities of gender, sexuality, race, and culture, simultaneously challenging some institutionalized attitudes (especially heterosexism) while reinforcing others (especially sexism and racism)--making the use of drag as a locus of discovery in both films, at best, ambivalent. PMID- 15132491 TI - Balancing acts: drag queens, gender and faith. AB - While engaged in research on the same-sex marriage debate in mainline denominations, I interviewed 23 LGBT Christians, four of whom were drag queens. While it is not possible to generalize from such a small sample, the drag queens in this study insist on maintaining their identity as Christians despite the hegemonic discourse that renders faith and LGBT identities mutually exclusive. They developed innovative approaches to reconciling their gender and sexual identities with their spirituality. Their innovations are potentially liberating not just for them personally, but for LGBT people generally because they challenge Christianity's rigid dichotomies of gender and sexuality. PMID- 15132490 TI - Racializing white drag. AB - While drag is primarily understood as a performance of gender, other performative categories such as race, class, and sexuality create drag meaning as well. Though other categories of identification are increasingly understood as essential elements of drag by performers of color, whiteness remains an unmarked category in the scholarship on drag performances by white queens. In this paper, I argue that drag by white queens must be understood as a performance of race as well as gender and that codes of gender excess are specifically constructed through the framework of these other axes of identity. This essay asks whether white performance by white queens necessarily reinscribes white supremacy through the performance of an unmarked white femininity, or might drag performance complicate (though not necessarily subvert) categories of race as well as gender? In this essay, I will suggest that camp drag performances, through the deployment of class as a crucial category of performative femininity, might indeed be a key site through which whiteness is denaturalized and its power challenged. Specifically, I will read on camp as a politicized mode of race, class and gender performance, focusing on the intersections of these categories of identity in the drag performance of Divine. PMID- 15132492 TI - A comparative analysis of hijras and drag queens: the subversive possibilities and limits of parading effeminacy and negotiating masculinity. AB - Gender studies in general and queer studies in particular have stressed the notion of imitation, play, and performance of gender. In this essay I undertake a comparative analysis of the hijra and the drag queen in terms of the shared and disparate subversive possibilities and limits of the gendered performances they undertake. Studying the hijra alongside the drag queen will in no way mean conflating the two categories. Rather, I explore the cultural nuances involved in the hijra performance, including its ritualistic and religious aspects. While my analysis relies heavily on previously written works about hijras and drag queens, I have also had the opportunity to meet and visit with hijras during several marriage ceremonies of cousins and other relatives from 1992 to 1998 in New Delhi. PMID- 15132493 TI - Beyond the boundaries of the classroom: teaching about gender and sexuality at a drag show. AB - Like much of the general public, the vast majority of my students strongly hold dichotomous, essentialist outlooks about what the categories female and male/gay and straight are supposed to represent and be. One way that I have found to challenge these oppressive worldviews, and also to queer my classes in the process, is to take my course participants to drag shows and/or to use videotapes of drag queens and drag kings in my classes. As part of an ongoing ethnography of drag performers I am undertaking, I have taken over 300 students to drag shows over the past eight years. Female students have often found attending a drag show to be a fun experience free of the sexual harassment found in most bars, while male students often contextually experience being a social minority for the first time in their life. From both attending drag shows and/or watching recordings of them I show in class, students have reported gaining an experiential appreciation of the performed basis of gender, sexuality, and inequality. Or, stated slightly differently, students begin to understand how the stratification system metaphorically makes drag queens of us all. This, in turn, provides the basis of the foremost argument I make in all my classes: equality will not be realized until nondichotomous, truly new ways of relating to others are envisioned and acted upon. PMID- 15132494 TI - Dynamic modeling of renal blood flow in Dahl hypertensive and normotensive rats. AB - A method is proposed in this paper which allows characterization of renal autoregulatory dynamics and efficiency using quantitative mathematical methods. Based on data from rat experiments, where arterial blood pressure and renal blood flow are measured, a quantitative model for renal blood flow dynamics is constructed. The mathematical structure for the dynamics is chosen as a "grey-box model," i.e. the model structure is inspired from physiology, but the actual parameters is found by numerical methods. Based on a number of experiments, features are extracted from the estimated parameters, which describe myogenic responses and tubuloglomerular feedback responses separately. The method is applied to data from normo- and hypertensive Dahl rats, and a discriminator that separates data from normotensive Dahl R rats and hypertensive Dahl S rats is constructed. PMID- 15132495 TI - Different pulse shapes to obtain small fiber selective activation by anodal blocking--a simulation study. AB - The aim of this study was to investigate whether it is possible to reduce a charge per pulse, which is needed for selective nerve stimulation. Simulation is performed using a two-part simulation model: a volume conductor model to calculate the electrical potential distribution inside a tripolar cuff electrode and a human fiber model to simulate the fiber response to simulation. Selective stimulation is obtained by anodal block. To obtain anodal block of large fibers, long square pulses (> 350 micros) with a relatively high currents (1-2.5 mA) are usually required. These pulses might not be safe for a long-term application because of a high charge per pulse. In this study, several pulse shapes are proposed that have less charge per pulse compared with the conventional square pulse and would therefore be safer in a chronic application. Compared with the conventional square pulse, it was possible to reduce the charge with all proposed pulse shapes, but the best results are obtained with a combination of a square depolarizing pulse and a blocking pulse. The charge per pulse was up to 32% less with that pulse shape than with a square pulse. Using a hyperpolarizing anodal prepulse preceding a square pulse, it was not possible to block nerve fibers in a whole nerve bundle and to obtain reduction of a charge per phase. Reduction of the charge could be achieved only with spatially selective blocking. The charge per phase was larger for the combination of a hyperpolarizing anodal prepulse and a two-step pulse than for the two-step pulse alone. PMID- 15132496 TI - Bayesian class discovery in microarray datasets. AB - A novel approach to class discovery in gene expression datasets is presented. In the context of clinical diagnosis, the central goal of class discovery algorithms is to simultaneously find putative (sub-)types of diseases and to identify informative subsets of genes with disease-type specific expression profile. Contrary to many other approaches in the literature, the method presented implements a wrapper strategy for feature selection, in the sense that the features are directly selected by optimizing the discriminative power of the used partitioning algorithm. The usual combinatorial problems associated with wrapper approaches are overcome by a Bayesian inference mechanism. On the technical side, we present an efficient optimization algorithm with guaranteed local convergence property. The only free parameter of the optimization method is selected by a resampling-based stability analysis. Experiments with Leukemia and Lymphoma datasets demonstrate that our method is able to correctly infer partitions and corresponding subsets of genes which both are relevant in a biological sense. Moreover, the frequently observed problem of ambiguities caused by different but equally high-scoring partitions is successfully overcome by the model selection method proposed. PMID- 15132497 TI - Adaptive BCI based on variational Bayesian Kalman filtering: an empirical evaluation. AB - This paper proposes the use of variational Kalman filtering as an inference technique for adaptive classification in a brain computer interface (BCI). The proposed algorithm translates electroencephalogram segments adaptively into probabilities of cognitive states. It, thus, allows for nonstationarities in the joint process over cognitive state and generated EEG which may occur during a consecutive number of trials. Nonstationarities may have technical reasons (e.g., changes in impedance between scalp and electrodes) or be caused by learning effects in subjects. We compare the performance of the proposed method against an equivalent static classifier by estimating the generalization accuracy and the bit rate of the BCI. Using data from two studies with healthy subjects, we conclude that adaptive classification significantly improves BCI performance. Averaging over all subjects that participated in the respective study, we obtain, depending on the cognitive task pairing, an increase both in generalization accuracy and bit rate of up to 8%. We may, thus, conclude that adaptive inference can play a significant contribution in the quest of increasing bit rates and robustness of current BCI technology. This is especially true since the proposed algorithm can be applied in real time. PMID- 15132498 TI - A novel method for determining the nature of time series. AB - The delay vector variance (DVV) method, which analyzes the nature of a time series with respect to the prevalence of deterministic or stochastic components, is introduced. Due to the standardization within the DVV method, it is possible both to statistically test for the presence of nonlinearities in a time series, and to visually inspect the results in a DVV scatter diagram. This approach is convenient for interpretation as it conveys information about the linear or nonlinear nature, as well as about the prevalence of deterministic or stochastic components in the time series, thus unifying the existing approaches which deal either with only deterministic versus stochastic, or the linear versus nonlinear aspect. The results on biomedical time series, namely heart rate variability (HRV) and functional Magnetic Resonance Imaging (fMRI) time series, illustrate the applicability of the proposed DVV-method. PMID- 15132499 TI - Characterization of event related potentials using information theoretic distance measures. AB - Analysis of event-related potentials (ERPs) using signal processing tools has become extremely widespread in recent years. Nonstationary signal processing tools such as wavelets and time-frequency distributions have proven to be especially effective in characterizing the transient phenomena encountered in event-related potentials. In this paper, we focus on the analysis of event related potentials collected during a psychological experiment where two groups of subjects, spider phobics and snake phobics, are shown the same set of stimulus: A blank stimulus, a neutral stimulus and a spider stimulus. We introduce a new approach, based on time-frequency distributions, for analyzing the ERPs. The difference in brain activity before and after a stimulus is presented is quantified using distance measures as adapted to the time-frequency plane. Three different distance measures, including a new information theoretic distance measure, are applied on the time-frequency plane to discriminate between the responses of the two groups of subjects. The results illustrate the effectiveness of using distance measures combined with time-frequency distributions in differentiating between the two classes of subjects and the different regions of the brain. PMID- 15132500 TI - EEG signal modeling using adaptive Markov process amplitude. AB - In this paper, an adaptive Markov process amplitude algorithm is used to model and simulate electroencephalogram (EEG) signals. EEG signal modeling is used as a tool to identify pathophysiological EEG changes potentially useful in clinical diagnosis. The least mean square algorithm is adopted to continuously estimate the parameters of a first-order Markov process model. EEG signals recorded from rodent brains during injury and recovery following global cerebral ischemia are utilized as input signals to the model. The EEG was recorded in a controlled experimental brain injury model of hypoxic-ischemic cardiac arrest. The signals from the injured brain during various phases of injury and recovery were modeled. Results show that the adaptive model is accurate in simulating EEG signal variations following brain injury. The dynamics of the model coefficients successfully capture the presence of spiking and bursting in EEG. PMID- 15132501 TI - A novel speech-processing strategy incorporating tonal information for cochlear implants. AB - Good performance in cochlear implant users depends in large part on the ability of a speech processor to effectively decompose speech signals into multiple channels of narrow-band electrical pulses for stimulation of the auditory nerve. Speech processors that extract only envelopes of the narrow-band signals (e.g., the continuous interleaved sampling (CIS) processor) may not provide sufficient information to encode the tonal cues in languages such as Chinese. To improve the performance in cochlear implant users who speak tonal language, we proposed and developed a novel speech-processing strategy, which extracted both the envelopes of the narrow-band signals and the fundamental frequency (F0) of the speech signal, and used them to modulate both the amplitude and the frequency of the electrical pulses delivered to stimulation electrodes. We developed an algorithm to extract the fundatmental frequency and identified the general patterns of pitch variations of four typical tones in Chinese speech. The effectiveness of the extraction algorithm was verified with an artificial neural network that recognized the tonal patterns from the extracted F0 information. We then compared the novel strategy with the envelope-extraction CIS strategy in human subjects with normal hearing. The novel strategy produced significant improvement in perception of Chinese tones, phrases, and sentences. This novel processor with dynamic modulation of both frequency and amplitude is encouraging for the design of a cochlear implant device for sensorineurally deaf patients who speak tonal languages. PMID- 15132502 TI - Electrical impedance tomography for imaging tissue electroporation. AB - Electroporation is a method to introduce molecules, such as gene constructs or small drugs, into cells by temporarily permeating the cell membrane with electric pulses. In molecular medicine and biotechnology, tissue electroporation is performed with electrodes placed in the target area of the body. Currently, tissue electroporation, as with all other methods of molecular medicine, is performed without real-time control or near-term information regarding the extent and degree of electroporation. This paper expands the work from our previous study by implementing new ex vivo experimental data with "front-tracking" analysis for the image reconstruction algorithm. The experimental data is incorporated into numerical simulations of electroporation procedures and images are generated using the new reconstruction algorithm to demonstrate that electrical impedance tomography (EIT) can produce an image of the electroporated area. Combining EIT with electroporation could become an important biotechnological and medical technique to introduce therapeutic molecules into cells in tissue at predetermined areas of the body. PMID- 15132503 TI - Spatio-temporal cortical source imaging of brain electrical activity by means of time-varying parametric projection filter. AB - In the present study, we explore suitable spatio-temporal filters for inverse estimation of an equivalent dipole-layer distribution from the scalp electroencephalogram (EEG) for imaging of brain electric sources. We propose a time-varying parametric projection filter (tPPF) for the spatio-temporal EEG analysis. The performance of this tPPF algorithm was evaluated by computer simulation studies. An inhomogeneous three-concentric-spheres model was used in the present simulation study to represent the head volume conductor. An equivalent dipole layer was used to represent equivalently brain electric sources and estimated from the scalp potentials. The tPPF filter was tested to remove time-varying noise such as instantaneous artifacts caused by eyes-blink. The present simulation results indicate that the proposed time-variant tPPF method provides enhanced performance in rejecting time-varying noise, as compared with the time-invariant parametric projection filter. PMID- 15132504 TI - Local contralateral subtraction based on bilateral symmetry of lung for reduction of false positives in computerized detection of pulmonary nodules. AB - A novel method called local contralateral subtraction has been developed for the removal of normal anatomic structures in chest radiographs based on the symmetry between the left and right lung regions. The method was oriented to the reduction of false positives reported by a computer-aided diagnosis (CAD) scheme for detection of lung nodules in chest radiographs. In our method, two regions of interest (ROIs) are extracted, one from the position where a nodule candidate is located, and the other from the anatomically corresponding location in the opposite lung, which contains similar normal structures. A wavelet-based, multiresolution image registration method is employed for matching the two ROIs, and subtraction is performed. If no structure remains in the subtracted ROI, then the original ROI is identified as negative (i.e., it contains only normal structures); otherwise, it is regarded as positive (i.e., it contains a nodule). A measure that quantifies the remaining structures was developed to distinguish between nodules and false positives. Application of the method to clinical chest radiographs showed that it was effective in eliminating normal anatomic structures and reducing the number of false detections in the CAD scheme for detection of lung nodules. PMID- 15132505 TI - Estimating joint contact areas and ligament lengths from bone kinematics and surfaces. AB - We present a novel method for modeling contact areas and ligament lengths in articulations. Our approach uses volume images generated by computed tomography and allows the in vivo and noninvasive study of articulations. In our method, bones are modeled both implicitly (scalar distance fields) and parametrically (manifold surfaces). Using this double representation, we compute interbone distances and estimate joint contact areas. Using the same types of representation, we model ligament paths; in our model, the ligaments are approximated by the shortest paths in a three-dimensional space with bone obstacles. We demonstrate the method by applying our contact area and ligament model to the distal radioulnar joints of a volunteer diagnosed with malunited distal radius fracture in one forearm. Our approach highlights focal changes in the articulation at the distal radioulnar joint (location and area of bone contact) and potential soft-tissue constraints (increased "length" of the distal ligaments and ligament-bone impingement in the injured forearm). Results suggest that the method could be useful in the study of normal and injured anatomy and kinematics of complex joints. PMID- 15132506 TI - Automatic segmentation of thalamus from brain MRI integrating fuzzy clustering and dynamic contours. AB - Thalamus is an important neuro-anatomic structure in the brain. In this paper, an automated method is presented to segment thalamus from magnetic resonance images (MRI). The method is based on a discrete dynamic contour model that consists of vertices and edges connecting adjacent vertices. The model starts from an initial contour and deforms by external and internal forces. Internal forces are calculated from local geometry of the model and external forces are estimated from desired image features such as edges. However, thalamus has low contrast and discontinues edges on MRI, making external force estimation a challenge. The problem is solved using a new algorithm based on fuzzy C-means (FCM) unsupervised clustering, Prewitt edge-finding filter, and morphological operators. In addition, manual definition of the initial contour for the model makes the final segmentation operator-dependent. To eliminate this dependency, new methods are developed for generating the initial contour automatically. The proposed approaches are evaluated and validated by comparing automatic and radiologist's segmentation results and illustrating their agreement. PMID- 15132507 TI - Multichannel magnetic stimulation system design considering mutual couplings among the stimulation coils. AB - We introduce some simulation and experiment results of the multichannel magnetic stimulator development that has been carried out as an initial attempt to realize a multichannel functional magnetic stimulator. For efficient functional magnetic stimulations, precise spatial localization of stimulation sites without any movements of the stimulation coils is very important. We have found that the mutual coupling effect among the adjacent stimulation coils in the coil array has to be considered in the determination of the charge voltages in some coil array configurations. Experimental results obtained with a 4-channel magnetic stimulator are presented. PMID- 15132508 TI - Pulse oximetry theory and calibration for low saturations. AB - Pulse oximetry is a widely used technique in biomedical optics, but currently available pulse oximeters rely on empirical calibration approaches, which perform poorly at low saturations. We present an exact solution for pulse oximetry and show how this can be used as the basis for the development of a semiempirical calibration approach that may be useful, especially at low saturations and variable probe geometries. This new approach was experimentally tested against traditional empirical calibration techniques on transmission pulse oximetry for monitoring of fetal sheep using a minimally invasive spiral probe. The results open the way for the development of more accurate pulse oximetry. PMID- 15132509 TI - A closed-loop mechanical ventilation controller with explicit objective functions. AB - A closed-loop lung ventilation controller was designed, aiming to: 1) track a desired end-tidal CO2 pressure (Pet CO2), 2) find the positive end-expiratory pressure (PEEP) of minimum estimated respiratory system elastance (Ers,e), and 3) follow objective functions conjectured to reduce lung injury. After numerical simulations, tests were performed in six paralyzed piglets. Respiratory mechanics parameters were estimated by the recursive least squares (RLS) method. The controller incorporated a modified PI controller for Pet CO2 and a gradient descent method for PEEP. In each animal, three automated PEEP control runs were performed, as well as a manual PEEP titration of Ers,e and a multiple PetCO2 step change trial. Overall performance indexes were obtained from PEEP control, such as minimum Ers,e (37.0 +/- 4.5 cmH2O x L(-1)), time to reach the minimum Ers,e (235 +/- 182 s) and associated PEEP (6.5 +/- 1.0 cmH2O), and from Pet CO2 control, such as rise time (53 +/- 22 s), absolute overshoot/undershoot of PetCO2 (3 +/- 1 mmHg), and settling time (145 +/- 72 s). The resulting CO2 controller dynamics approximate physiological responses, and results from PEEP control were similar to those obtained by manual titration. Multiple dependencies linking the involved variables are discussed. The present controller can help to implement and evaluate objective functions that meet clinical goals. PMID- 15132510 TI - A fully integrated neural recording amplifier with DC input stabilization. AB - This paper presents a low-power low-noise fully integrated bandpass operational amplifier for a variety of biomedical neural recording applications. A standard two-stage CMOS amplifier in a closed-loop resistive feedback configuration provides a stable ac gain of 39.3 dB at 1 kHz. A subthreshold PMOS input transistor is utilized to clamp the large and random dc open circuit potentials that normally exist at the electrode-electrolyte interface. The low cutoff frequency of the amplifier is programmable up to 50 Hz, while its high cutoff frequency is measured to be 9.1 kHz. The tolerable dc input range is measured to be at least +/- 0.25 V with a dc rejection factor of at least 29 dB. The amplifier occupies 0.107 mm2 in die area, and dissipates 115 microW from a 3 V power supply. The total measured input-referred noise voltage in the frequency range of 0.1-10 kHz is 7.8 microVrms. It is fabricated using AMI 1.5 microm double-poly double-metal n-well CMOS process. This paper presents full characterization of the dc, ac, and noise performance of this amplifier through in vitro measurements in saline using two different neural recording electrodes. PMID- 15132511 TI - Three-dimensional localization of cochlear implant electrodes using epipolar stereophotogrammetry. AB - Three-dimensional (3-D) localization of individual cochlear implant electrodes within the inner ear is of importance for modeling the electrical field of the cochlea, designing the electrode array, and programming the associated speech processor. A 3-D reconstruction method of cochlear implant electrodes is proposed to localize individual electrodes from two X-ray views in combination with the spiral computed tomography technique. By adapting epipolar geometry to the configuration of an X-ray imaging system, we estimate individual electrode locations in the least square sense without using a patient attachment required by an existing stereophotogrammetry technique. Furthermore, our method does not require any knowledge of the intrinsic and extrinsic parameters of the imaging system. The performance of our method is studied in numerical simulation and with patient data and is found to be sufficiently accurate for clinical use. The maximum root mean-square errors measured are 0.0445 and 0.214 mm for numerical simulation and patient data, respectively. PMID- 15132512 TI - Comparative psychometric analysis of vector and isochrone cardiac activation maps. AB - Isochronal cardiac activation maps can be constructed from local activation times associated with spatial locations, and are frequently used to study cardiac arrhythmias. Cardiac velocity vector mapping has been proposed as an alternative method to study cardiac activation in both clinical and research environments. Velocity vectors inherently contain more information than scalar measures of latency, but it is unknown how vector maps and isochronal maps compare when they are used to identify patterns and features associated with arrhythmias. In order to quantitatively compare these two visualization methods, eight cardiologists were asked to complete forced-choice tasks in which they selected ablation sites based on synthetic vector or isochronal maps. Maps varied in arrhythmia complexity, number of vectors or activation times included, and errors in magnitude or angle for maps of velocity vectors. Quantitative comparison was achieved by using psychometric functions to characterize the learning curve and the total number of measurements needed in order to choose a correct ablation site. For simple arrhythmias, performance with vector maps was superior to isochronal maps. Subjects required fewer measurements, and learned more rapidly by studying vector maps. For more complex arrhythmias, there was no significant difference in performance between vector and isochronal maps. However, arrhythmia features were clearer with vector maps, even though this clarity did not necessarily change the ablation site choice. When errors were added to vector maps, performance was satisfactory for angle errors < 55 degrees, and speed errors did not affect performance. PMID- 15132513 TI - Proposed corrections for the quantification of coupling patterns by recurrence plots. AB - The aim of this paper is to quantify the coupling during phase-locking patterns by using the recurrence plot quantification approach. We found that the percent determinism of the recurrences and the entropy of recurrences--corrected for border effect induced by finite data lenght--succeeded in distinguishing three coupling conditions both in simulation signals and in real data from cardiorespiratory synchronization experiments. PMID- 15132514 TI - Cultured neurons coupled to microelectrode arrays: circuit models, simulations and experimental data. AB - The purpose of this paper is to characterize the neuron-microelectrode junction, based on the equivalent electric-circuit approach. As a result, recording of action potentials can be simulated with a general-purpose circuit simulation program such as HSPICE. The response of the microelectrode was analyzed as a function of parameters such as sealing resistance and adhesion conditions. The models of the neuron and microelectrode implemented in HSPICE were first described. These models were used to simulate the behavior of the junction between a patch of neuronal membrane (described by the compartmental model) and a microelectrode. PMID- 15132515 TI - A novel method of estimation of DPOAE signals. AB - A new method of measurement of distortion product otoacoustic emission (DPOAE) signal level based on a recently introduced nonlinear adaptive method of extraction of nonstationary sinusoids is presented. Essentially, three units of such an algorithm are employed to extract and measure the two stimuli and the DPOAE signal. Each unit has the capability of locking on a specified sinusoidal component of the input signal and tracking its variations over time. Performance of the proposed method is demonstrated with the aid of computer simulations and is verified in laboratory using recorded clinical data. Comparison is made between the proposed technique and existing methods. The proposed method features structural simplicity which renders it particularly attractive for implementation on both software and hardware platforms. It offers a high degree of immunity with regard to background noise and parameter variations. Compared to conventional methods, the proposed method offers a shorter measurement time which is of significant value in clinical examinations. PMID- 15132516 TI - Interictal spike detection using the Walsh transform. AB - The objective of this study was to evaluate the feasibility of using the Walsh transformation to detect interictal spikes in electroencephalogram (EEG) data. Walsh operators were designed to formulate characteristics drawn from experimental observation, as provided by medical experts. The merits of the algorithm are: 1) in decorrelating the data to form an orthogonal basis and 2) simplicity of implementation. EEG recordings were obtained at a sampling frequency of 500 Hz using standard 10-20 electrode placements. Independent sets of EEG data recorded on 18 patients with focal epilepsy were used to train and test the algorithm. Twenty to thirty minutes of recordings were obtained with each subject awake, supine, and at rest. Spikes were annotated independently by two EEG experts. On evaluation, the algorithm identified 110 out of 139 spikes identified by either expert (True Positives = 79%) and missed 29 spikes (False Negatives = 21%). Evaluation of the algorithm revealed a Precision (Positive Predictive Value) of 85% and a Sensitivity of 79%. The encouraging preliminary results support its further development for prolonged EEG recordings in ambulatory subjects. With these results, the false detection (FD) rate is estimated at 7.2 FD per hour of continuous EEG recording. PMID- 15132517 TI - Psychological functioning in cancer patients treated with radiotherapy. AB - Although many side-effects of radiotherapy (RT) are physical, previous studies have shown that patients, when treated with RT, also experience psychological problems. This review describes the psychological functioning of cancer patients prior to, during, and after RT. Moreover, we examined whether medical factors are related to psychological functioning. Using Medline and Psychlit databases over the period 1980-2002, 45 articles were studied for this review. Although some global trends were found, studies revealed great variability in psychological functioning prior to, during, and after RT. Longitudinal studies should include pre-, during, and post-RT assessments of psychological functioning and also take into account the previous psychological stability of the patient. PMID- 15132518 TI - Male sexuality after cancer treatment--needs for information and support: testicular cancer compared to malignant lymphoma. AB - Testicular cancer (TC) as well as malignant lymphoma (ML), both have nowadays an excellent prognosis. However, both types of cancer may be diagnosed at young adulthood and patients may experience sexual concerns. In this article the need for information and support concerning sexuality will be explored, and the traumatic impact of cancer diagnosis with respect to this will be considered. A total of 264 patients with testicular cancer, median age 36 (S.D. 9.7) years, and 50 patients with malignant lymphoma, median age 42 (S.D. 11.7) years returned a questionnaire concerning sexual functioning; four items assessed the need for information or support concerning sexuality, at diagnosis and at follow-up. It appeared that more than half of the patients with testicular cancer reported a lack of information and support concerning sexuality during treatment; 67% of them still had a need for information at follow-up. These rates were significantly lower for patients with malignant lymphoma. Especially patients with testicular cancer who suffered sexual dysfunction reported extremely high needs for information and support. According to these findings it can be concluded that more attention should be paid to the doctor-patient communication with respect to sexual concerns in general, and especially where it concerns patients with testicular cancer. PMID- 15132519 TI - Diabetes rehabilitation: development and first results of a Multidisciplinary Intensive Education Program for patients with prolonged self-management difficulties. AB - For a number of diabetes patients regular care may be insufficient. A Multidisciplinary Intensive Education Program (MIEP), based on the empowerment approach, has been developed to help patients obtain their treatment goals (adequate self-management, glycemic control and quality of life). The aim of this pilot study is to determine the effects of MIEP and it's mechanisms of influence. MIEP consisted of 12 days group-sessions and individual counseling. At baseline and 3-months follow-up, blood-glucose (HbA1c), quality of life, health locus of control, distress, and knowledge were obtained (N = 51). Paired T-tests and regression analyses were conducted. HbA1c, and knowledge improved significantly, patients rated themselves healthier and were more internal and less powerful others oriented. Baseline scores explained effects in HbA1c, and quality of life. Locus of control significantly contributed in effects on quality of life. MIEP benefited patients with prolonged self-management difficulties, and this form of care seems to complement regular care. PMID- 15132520 TI - Qualitative study investigating the process of giving anti-smoking advice in general practice. AB - General practitioners' (GPs') anti-smoking advice promotes patients' smoking cessation but little is known about how GPs use their short consultations to give advice. We used semi-structured interviews with 27 UK GPs to investigate how GPs believe they should advise smokers to stop and the reasons underpinning these beliefs. GPs reported a limited repertoire of techniques for dealing with smokers who were not motivated to stop. They also reported using confrontational advice giving styles with patients who continued to smoke despite suffering from smoking related illnesses. GPs might find it easier and more rewarding to discuss smoking with patients if they possessed a greater range of skills for dealing with non motivated smokers. PMID- 15132521 TI - Dutch dentists' views of informed consent: a replication study. AB - The aim of this study was to replicate a previous study on informed consent in dental practice, because of the low response-rate of that study. The present study assessed Dutch dentists' knowledge, their attitudes and self-efficacy towards the principle of informed consent, by means of a strongly shortened version of the questionnaire used in the original study. This questionnaire was sent to 384 Dutch dental practitioners; 60.2% of them responded. The results obtained in this study are highly comparable with the results of the original study, thereby increasing the confidence in the outcomes. Results of both studies indicate that dentists still have problems with some aspects of informed consent. Specifically, dentists' fear for legal procedures and the difficulty they have with informing immigrant patients warrants further attention. PMID- 15132522 TI - How do patients with rheumatic disease experience their relationship with their doctors? A qualitative study of experiences of stress and support in the doctor patient relationship. AB - This study is evaluated what patients with rheumatic disease perceive as important in their medical encounters. We interviewed two groups of patients: one with a well-defined inflammatory condition (rheumatoid arthritis (RA) or ankylosing spondylitis) (n = 12) and one with non-inflammatory widespread chronic pain such as fibromyalgia (n = 14). Both groups focused on their relationship to their doctor. Two central themes emerged as of importance: 'to be seen' and 'to be believed'. However, these themes had different connotations for the two groups. For the patients with inflammatory conditions, 'to be seen' implied being seen as an individual and not as a mere diagnosis, and 'to be believed' as far as pain and suffering were concerned. For patients with non-inflammatory chronic pain 'to be seen' and 'to be believed' primarily implied being able to obtain a useful somatic diagnosis. Practical implications of these findings are discussed. PMID- 15132523 TI - Written orthopedic patient education materials from the point of view of empowerment by education. AB - Written patient education materials are one way of supporting patient empowerment. The aim of this study was to evaluate written orthopedic patient education materials (n = 25) drawn from a university hospital's electronic databank. In the absence of suitable tools for evaluation, an analytic framework was developed by a panel of nursing scientists and clinical experts. The materials were evaluated from the point of view of empowerment by their external appearance, content and instructiveness. In terms of their external appearance the materials were well prepared, but much was left to be desired with regard to contents and instructiveness. Development needs to focus on making better use of methods of visual representation, on increasing the coverage of content areas that so far have received less attention, like social, experiential, ethical and financial empowerment, and on providing a more focused perspective on the patient. PMID- 15132524 TI - Evaluation of nutritional education using concept mapping. AB - The concept mapping method is presented in the current study as a new tool to assess the learning process taking part in the hallmark of a nutritional education program addressed to obese diabetic patients. POPULATION: eight patients were interviewed prior to and after completion of 1-week in-hospital stay during which concept maps were designed. Concept maps quantitative and qualitative analysis disclose both (i) the importance of previous knowledge among patients prior to nutritional education and (ii) the maintenance of misconceptions after it. Nutritional education allows patients to acquire and structure their knowledge while providing them with a certain amount of medical vocabulary. An underlying correlation between concept maps design and the results of psychological tests identifying eating behaviour troubles (EBT), depression or anxiety has not been clearly identified. However, the nutritional education is more beneficial to those patients with a higher degree of self-assertiveness and with a lesser degree of anxiety, depression and eating disorder. PMID- 15132525 TI - Effects of an empowerment-based psychosocial intervention on quality of life and metabolic control in type 2 diabetic patients. AB - This study was aimed at determining impact of an empowerment-based psychosocial intervention on the patients' quality of life and glycemic control as compared to patients in standard care. Consecutively recruited type 2 diabetic patients, scheduled for their regular medical check-ups, were individually acquainted with empowerment-based principles and invited to participate in an empowering psychosocial course. The response rate was 35% giving a number of 73 patients who were treated in eight separate groups. The treated patients reported their quality of life to be improved after the course regarding its psychological and social aspects. Their glycemic control also improved and remained so after 3- and 6-month follow-up periods, still being in a category of poor control. Empowerment based psychosocial intervention in type 2 diabetic patients was shown to favourably affect their quality of life and to improve their metabolic control, the latter in a modest degree. Better educated patients believing in internal health control and efficacy of diabetes treatment seemed to benefit the most. Further research is needed in order to highlight individual preferences for different educational approaches, as well as social and cultural factors affecting them. PMID- 15132526 TI - The Patient Perspective Survey (PPS): a new tool to improve consultation outcome and patient involvement in general practice patients with complex health problems. Psychometric testing and development of a final version. AB - The Patient Perspective Survey (PPS) is designed to enhance consultation outcome and patient participation among general practice patients with complex health problems. This article presents the final version of the survey. This version is more condensed, tested for psychometric and clinical properties, and with increased emphasis on coping. The development and testing was performed in two phases with two groups of GPs and patients. Indications for use and criteria for rejection of items were defined, based on comprehensive data analysis and clinical judgement. The number of items was reduced to 38 items. The majority of diagnoses were related to psychosocial or musculoskeletal disorders. For the survey's main dimensions (psychological and somatic items, everyday life situation and coping) sound psychometric properties were identified. Seven open ended questions were found useful for obtaining important new information and stimulating processes, often related to coping aspects. The revised 38-item PPS represents a condensed and final version, with increased focus on coping, and sound psychometric properties. PMID- 15132527 TI - The development and preliminary evaluation of a decision aid based on decision analysis for two treatment conditions: benign prostatic hyperplasia and hypertension. AB - This paper discusses the development and evaluation of a computerised decision aid that provides individualised information about Benign Prostatic Hyperplasia (BPH) and Hypertension to patients. The program is based on decision analysis, using decision trees as a way of providing users with information regarding the probability of different outcomes occurring, obtaining an individual evaluation of the different outcomes, before providing guidance on what might be the 'best' option for that patient. It is intended that the program can be used as the basis for helping patients to become more involved in decisions about their medical treatment. Eight health care professionals and 19 patients (9 with BPH and 10 with Hypertension) evaluated the program. Overall it was assessed positively by both health care professionals and patients. However, before it can be integrated into health care practice, the program is to be evaluated further in a randomised trial. PMID- 15132528 TI - From the design of a chiral Lewis acid catalyst to metal-catalyzed coupling reactions. AB - In this perspective, I describe my group's nonobvious path from an interest in chiral Lewis acid catalysis to a project focused on the development of new palladium and nickel catalysts for carbon-carbon bond-forming reactions. PMID- 15132530 TI - Exo- and endo-receptors in one. A novel class of supramolecular structures housing transition-metal-binding bi- and terpyridine units alongside lithium ion selective trispirotetrahydrofuranyl components. AB - The preparation of bipyridine and terpyridine ligands covalently linked via acetylenic and alkoxy tethers to rigid inositol orthoformate platforms is described. The constitution of compounds 3-6 is such that latent exo- and endo receptor properties are simultaneously present with specific binding sites for Li(+) and transition-metal ions. The 2-fold complexation in different regions is not dependent on prior molecular association. To gauge this unprecedented property, the model systems 11-14 were synthesized and their response to CuCl and FeCl(2) probed by UV-vis and MS techniques. In a similar fashion, the ability of 3-6 to ligate lithium ions chemoselectively was evaluated by electrospray methods. The introduction of controlled amounts of both types of cation followed, and conversion to insoluble oligomeric products presumed to result from integral incorporation of the different metal ions in the expected fashion was achieved. PMID- 15132529 TI - Single-site mutation and secondary structure stability: an isodesmic reaction approach. The case of unnatural amino acid mutagenesis Ala-->Lac. AB - A method is described to evaluate backbone interactions in proteins via computational unnatural amino acid mutagenesis. Several N-acetyl polyalanyl amides (AcA(n)NH(2)) were optimized in the representative helical (3(10)-, 4(13) , and a "hybrid" kappa-helix, n = 7, 9, 10, 14) and hairpin (two- and three stranded antiparallel beta-sheets with type I turns betaalphaalphaepsilon, n = 6, 9, 10) conformations, and extended conformers of N-acetyl polyalanyl methylamides (n = 2, 3) were used to derive multistranded beta-sheet fragments. Subsequently, each residue of every model structure was substituted, one at a time, with l lactic acid. The resulting mutant structures were again optimized, and group transfer energies DeltaE(GT) were obtained as heats of the isodesmic reactions: AcA(n)NHR + AcOMe --> AcA(x)LacA(y)NHR + AcNHMe (R = H, CH(3)). These group transfer energies correlate with the degree of charge polarization of the substituted peptide linkages as measured by the difference Deltae in H and O Mulliken populations in HN-C=O and with the H-bond distances in the "wild-type" structures. A good correlation obtains for the HF/3-21G and B3LYP/6-31G* group transfer energies. The destabilization effects are interpreted in terms of loss of interstrand and intrastrand H-bonds, decrease in Lewis basicity of the C=O group, and O...O repulsion. On the basis of several comparisons of Ala --> Lac DeltaE(GT)'s with heats of the NH --> CH(2) substitutions, the latter contribution is estimated (B3LYP/6-31G*) to range between 1.5 and 2.4 kcal mol( 1), a figure close to the recent experimental DeltaDeltaG(o) value of 2.6 kcal mol(-1) (McComas, C. C.; Crowley, B. M.; Boger, D. L. J. Am.Chem. Soc. 2003, 125, 9314). The partitioning yields the following maximum values of the electronic association energy of H-bonds in the examined sample of model structures (B3LYP/6 31G* estimates): 3(10)-helix D(e) = -1.7 kcal mol(-1), alpha-helix D(e) = -3.8 kcal mol(-1), beta-sheet D(e) = -6.1 kcal mol(-1). The premise of experimental evaluations of the backbone-backbone H-bonding that Ala --> Lac substitution in proteins is isosteric (e.g., Koh, J. T.; Cornish, V. W.; Schultz, P. G. Biochemistry 1997, 36, 11314) is often but not always corroborated. Examination of the integrity of H-bonding pattern and phi(i), psi(i) distribution identified several mutants with significant distortions of the "wild-type" structure resulting inter alia from the transitions between i, i + 3 and i, i + 4 H-bonding in helices, observed previously in the crystallographic studies of depsipeptides (Ohyama, T.; Oku, H.; Hiroki, A.; Maekawa, Y.; Yoshida, M.; Katakai, R. Biopolymers 2000, 54, 375; Karle, I. L.; Das, C.; Balaram, P. Biopolymers 2001, 59, 276). Thus, the isodesmic reaction approach provides a simple way to gauge how conformation of the polypeptide chain and dimensions of the H-bonding network affect the strength of backbone-backbone C=O...HN bonds. The results indicate that the stabilization provided by such interactions increases on going from 3(10)-helix to alpha-helix to beta-sheet. PMID- 15132531 TI - Unambiguous detection of target DNAs by excimer-monomer switching molecular beacons. AB - A new class of molecular beacons were developed in which pyrene fluorophores were connected both at 3' and 5' ends of a single-stranded oligonucleotide. The two pyrene-based fluorophores were synthesized from the same starting material, so that the preparation of the beacons was simplified. The detection strategy of the beacons for target DNAs is based on "excimer-monomer emission switching" of the pyrene fluorophores: excimer emission of the pyrene moieties changed to monomer one when the beacons hybridized with the targets. This type of two-state mode of fluorescence allows unambiguous detection of the target DNAs because strict 1:1 correlation between the nonhybridized and the hybridized beacons can be monitored by the presence of isoemissive points of the fluorescence changes. The beacons can detect target 19-mer DNAs and can discriminate the targets from their single nucleotide mismatches at 1 nM concentration. Advantages of the excimer-monomer switching molecular beacons were discussed in comparison with conventional ones. PMID- 15132532 TI - Metal ion catalysis in the beta-elimination reactions of N-[2-(4 pyridyl)ethyl]quinuclidinium and N-[2-(2-pyridyl)ethyl]quinuclidinium in aqueous solution. AB - Catalysis of the beta-elimination reaction of N-[2-(4 pyridyl)ethyl]quinuclidinium (1) and N-[2-(2-pyridyl)ethyl]quinuclidinium (2) by Zn(2+) and Cd(2+) in OH(-)/H(2)O (pH = 5.20-6.35, 50 degrees C, and mu = 1 M KCl) has been studied. In the presence of Zn(2+), the elimination reactions of both isomers occur from the Zn(2+)-complexed substrates (C). The equilibrium constants for the dissociation of the Zn(2+)-complexes are as follows: K(d) = 0.012 +/- 0.003 M (isomer 1) and K(d) = 0.065 +/- 0.020 M (isomer 2). The value of k(C)(H2O) for isomer 1 is 4.81 x 10(-6) s(-1). For isomer 2 both the rate constants for the "water" and OH(-)-induced reaction of the Zn(2+)-complexed substrate could be measured, despite the low concentration of OH(-) in the investigated reaction mixture [k(C)H2O)= 1.97 x 10(-6) s(-1) and k(C)(OH-)= 21.9 M(-1) s(-1), respectively]. The measured metal activating factor (MetAF), i.e., the reactivity ratio between the complexed and the uncomplexed substrate, is 8.1 x 10(4) for the OH(-)-induced elimination of 2. This high MetAF can be compared with the corresponding proton activating factor (Alunni, S.; Conti, A.; Palmizio Errico, R. J. Chem. Soc., Perkin Trans. 2 2000, 453), PAF = 1.5 x 10(6) and is in agreement with an E1cb irreversible mechanism (A(xh)D(E)* + D(N)) (Guthrie, R. D.; Jencks, W. P. Acc. Chem. Res. 1989, 22, 343). A value of k(C)(H2O)>or= 23 x 10(-7) s(-1) is estimated for the Cd(2+)-complexed isomer 2, while catalysis by Cd(2+) has not been observed for isomer 1. PMID- 15132534 TI - Reactions of sterically congested 1,5-hexadienes: Ab initio and DFT calculations on the competition between cope rearrangements and disrotatory cyclobutene ring opening reactions of bridged syn-tricyclo[4.2.0.0(2,5)]octa-3,7-dienes. AB - The thermolytic behavior of four syn-tricyclo[4.2.0.0(2,5)]octa-3,7-dienes, each spanned by four propano bridges (13, 14, 21, and 26), has been investigated by means of calculations at the UB3LYP/ 6-31G* and CASPT2/6-31G levels. These calculations predict that 13 should undergo a degenerate Cope rearrangement (E(A) = 19.6 kcal/mol), whereas the other three C(20)H(24) isomers should prefer a necessarily disrotatory cyclobutene ring-opening reaction. In the case of 14, the ring-opening reaction (E(A) = 27.2 kcal/mol) is concerted and leads directly to 15, a 4-fold bridged cyclooctatetraene. In the ring opening of 21, the 1,6-bridge in the 4-fold bridged bicyclo[4.2.0]octa-2,4,7-triene 31 prevents formation of the corresponding cyclooctatetraene. In the ring opening of 26, the 4-fold bridged bicyclo[4.2.0]octa-2,4,7-triene derivative 36 is predicted to form the corresponding bridged cyclooctatetraene 38, which should undergo bond shift. The results of these calculations are found to be in very good agreement with the results of experiments on these hydrocarbons. PMID- 15132533 TI - Formal radical cyclization onto benzene rings: a general method and its use in the synthesis of ent-nocardione A. AB - An indirect method is described for effecting radical cyclization onto a benzene ring. Cross-conjugated dienones 6, which are readily prepared from phenols, undergo radical cyclization (6 --> 7 --> 8), and the products (8) are easily aromatized. The method has been applied to the synthesis of ent-nocardione A (21). PMID- 15132535 TI - Direct preparation of allylic zirconium reagents from zirconocene-olefin complexes and alkenes. AB - A novel method for preparation of allylic zirconium reagents directly from 1 alkenes via zirconocene-olefin complex has been developed. Selective transfer of the hydride of zirconocene allyl hydride complex, a tautomer of zirconocene olefin complex, to diisopropyl ketone generates the corresponding zirconocene alkoxide allyl. The allylic zirconium reagents formed effects stereoselective allylation of aldehyde at 25 degrees C and -78 degrees C to provide syn- and anti homoallyl alcohols, respectively. The anti-isomer is formed via a six-membered chair transition state under kinetic control. The syn-selectivity can be rationalized by considering isomerization of the anti-adduct by a retroallylation process. PMID- 15132536 TI - Multigram-scale syntheses, stability, and photoreactions of A2A adenosine receptor antagonists with 8-styrylxanthine structure: potential drugs for Parkinson's disease. AB - The improved multigram-scale syntheses of the important 8-styrylxanthine A(2A) adenosine receptor antagonist MSX-2 (8), its water-soluble prodrug MXS-3 (9), and KW-6002 (16) are described. N-Alkylation reactions at different positions of uracil derivatives were optimized. Two different methods for xanthine formation from 6-amino-5-cinnamoylaminouracil precursors were investigated, (a) the elimination of water by alkaline catalysis and (b) hexamethyldisilazane as a condensing agent; the latter was found to be superior. The photosensitivity of 8 styrylxanthines was studied. The (E)-configurated stryrylxanthine MSX-2 (8) isomerized in diluted solution, and the resulting (Z)-isomer (10a) was isolated and characterized. Furthermore, we describe for the first time that solid 8 styrylxanthines can dimerize upon exposition to daylight or irradiation with UV light. The resulting cyclobutane derivatives with head-to-tail (syn) configuration exhibited a considerably lower A(2A) adenosine receptor affinity than their parent compounds. The dimerization product of MSX-2 was a moderately potent nonselective A(1) and A(2A) antagonist (K(i)(A(1)) = 273 nM, K(i) (A(2A)) = 175 nM) while the dimer of the related compound KW-6002 was inactive at A(1) and only weakly active at A(2A) adenosine receptors (K(i) = 1.57 microM). The light sensitivity of 8-styrylxanthine derivatives, not only in solution, but also in the solid state, has to be considered when using those compounds as pharmacological tools or drugs. PMID- 15132537 TI - Suspension ring-opening metathesis polymerization: the preparation of norbornene based resins for application in organic synthesis. AB - A series of norbornene-based resin beads were obtained by aqueous suspension ring opening metathesis polymerization (ROMP) and used as polymeric supports for organic synthesis. These resins were prepared from norbornene, norborn-2-ene-5 methanol, and cross-linkers such as bis(norborn-2-ene-5-methoxy)alkanes, di(norborn-2-ene-5-methyl)ether, and 1,3-di(norborn-2-ene-5-methoxy)benzene. The resulting unsaturated ROMP (U-ROMP) resins containing olefin repeat units were chemically modified using hydrogenation, hydrofluorination, chlorination, and bromination reactions to produce saturated ROMP resins with different chemical and physical properties. The hydrogenated ROMP (H-ROMP) resin was found to be highly resistant to acidic, basic, Lewis acid, and Birch reduction conditions and was assessed as a polymeric support in a series of solid-phase synthetic applications. The H-ROMP resin was found to have superior performance compared to polystyrene-divinylbenzene (PS-DVB) copolymers in aromatic nitration and acylation reactions. In a conventional five-step solid-phase synthesis of a hydantoin, similar results were obtained for both the H-ROMP and PS-DVB resins. The U-ROMP resin was also shown to be effective in the solid-phase syntheses of benzimidazoles and benzimidazolones. PMID- 15132538 TI - Stereospecific synthesis of optically active phenylpropylene oxides. AB - The stereospecific lithiation of diastereomeric phenylpropylene oxides has been studied as well as the trapping reaction with electrophiles. The reduction of the cis-alpha-benzoylpropylene oxide to give prevalently the anti-epoxy alcohol has been investigated as well. PMID- 15132539 TI - Palladium-catalyzed regioselective hydrodebromination of dibromoindoles: application to the enantioselective synthesis of indolodioxane U86192A. AB - A novel approach to the selective preparation of 4-bromoindoles was developed via Pd(OAc)(2)/rac-BINAP catalytic reactions. A variety of 4,6-dibromoindoles were transformed to 4-bromoindoles with high regioselectivity. This methodology, along with C-N and C-O bond-forming reactions developed in our laboratory, was applied to the enantioselective synthesis of indolodioxane U86192A, an antihypertensive agent. PMID- 15132540 TI - Application of ionic liquid halide nucleophilicity for the cleavage of ethers: a green protocol for the regeneration of phenols from ethers. AB - We have used the high nucleophilicity of bromide ion in the form of the ionic liquid, 1-n-butyl-3-methylimidazolium bromide ([bmim][Br]), for the nucleophilic displacement of an alkyl group to regenerate a phenol from the corresponding aryl alkyl ether. Using 2-methoxynaphthalene (1) as a model compound, we found that the combination of ionic liquid [bmim][Br] and p-toluenesulfonic acid with warming effected demethylation in 14 h, affording the desired product 2-naphthol (2) in good yield (97%). Various other protic acids (MsOH, hydrochloric acid (35%), dilute sulfuric acid (50%)) could be used as a proton source in this demethylation reaction. Under the same conditions, cleavage of alkyl alkyl ether 2-(3-methoxypropyl)naphthalene yielded mixture of corresponding 2-(3 bromopropyl)naphthalene and 2-(3-hydroxypropyl)naphthalene. Dealkylation of various aryl alkyl ethers could also be achieved using significantly reduced (i.e., stoichiometric) amounts of concentrated hydrobromic acid (47%) in the ionic liquid. Both procedures afforded the desired products in moderate to good yield; however, cleavage of aryl alkyl cyclic ether, 2,3-dihydrobenzofuran, resulted in low yield of the desired product o-2-bromoethylphenol. The convenience of this method for ether cleavage and its effectiveness using only a moderate excess of hydrobromic acid make it attractive as a green chemical method. PMID- 15132541 TI - Microwave-promoted and chelation-controlled double arylations of terminal olefinic carbon of vinyl ethers. AB - Herein we report a rapid, palladium-catalyzed terminal diarylation of the chelating olefin N,N-dimethyl(2-ethenyloxy)ethanamine under noninert conditions utilizing controlled microwave heating as a convenient energy source. Among the aryl bromides examined, both electron-rich and electron-poor substrates were demonstrated to furnish useful yields after only 10-120 min of directed microwave heating at 160-200 degrees C. The good terminal regioselectivity suggests that the precatalyst (Herrmann's palladacycle) serves as a source of weakly coordinated palladium(0) in the investigated high-temperature Heck process. PMID- 15132542 TI - Silvestrol and episilvestrol, potential anticancer rocaglate derivatives from Aglaia silvestris. AB - Two cytotoxic rocaglate derivatives possessing an unusual dioxanyloxy unit, silvestrol (1) and episilvestrol (2), were isolated from the fruits and twigs of Aglaia silvestris by bioassay-guided fractionation monitored with a human oral epidermoid carcinoma (KB) cell line. Additionally, two new baccharane-type triterpenoids, 17,24-epoxy-25-hydroxybaccharan-3-one (3) and 17,24-epoxy-25 hydroxy-3-oxobaccharan-21-oic acid (4), as well as eleven known compounds, 1beta,6alpha-dihydroxy-4(15)-eudesmene (5), ferulic acid (6), grasshopper ketone (7), apigenin, cabraleone, chrysoeriol, 1beta,4beta-dihydroxy-6alpha,15alpha epoxyeudesmane, 4-hydroxy-3-methoxyacetophenone, 4-hydroxyphenethyl alcohol, ocotillone, and beta-sitosterol 3-O-beta-D-glucopyranoside, were also isolated and characterized. The structures of compounds 1-4 were elucidated by spectroscopic studies and by chemical transformation. The absolute stereochemistry of silvestrol (1) was established by a X-ray diffraction study of its di-p-bromobenzoate derivative, and the structure of 3 was also confirmed by single-crystal X-ray diffraction. The isolates and chemical transformation products were evaluated for cytotoxicity against several human cancer cell lines, and silvestrol (1) and episilvestrol (2) exhibited potent in vitro cytotoxic activity. Silvestrol (1) was further evaluated in vivo in the hollow fiber test and in the murine P-388 leukemia model. PMID- 15132543 TI - Kinetics and mechanism of hydrolysis of N-acyloxymethyl derivatives of azetidin-2 one. AB - The pH-independent, acid-catalyzed and base-catalyzed hydrolyses of N acyloxymethylazetidin-2-ones all occur at the ester function. The pH-independent hydrolysis involves rate-limiting alkyl C-O fission and formation of an exocyclic beta-lactam iminum ion. This iminium ion is then trapped by water at the exocyclic iminium carbon atom, rather than at the beta-lactam carbonyl carbon atom, to form the corresponding N-hydroxymethylazetidin-2-ones. Calculations carried out at the B3LYP/6-31+G(d) level of theory also support that nucleophilic attack by water takes place at the exocyclic carbon rather than at the beta lactam carbonyl carbon of the iminium ion. The mechanism for the acid-catalyzed pathway involves a preequilibrium protonation, probably at the beta-lactam nitrogen, followed by rate-limiting alkyl C-O fission with formation of an exocyclic iminum ion. The base-catalyzed hydrolysis involves rate-limiting hydroxide attack at the ester carbonyl carbon. These results imply formation of a beta-lactam system containing a positively charged amide nitrogen atom that hydrolyzes via a pathway that preserves the beta-lactam structure in the product and provide further evidence that cleavage of the beta-lactam C-N bond is not as facile as is commonly imagined. PMID- 15132544 TI - Total synthesis and preliminary biological evaluation of cis-solamin isomers. AB - An efficient total synthesis of cis-solamin (1) has been achieved in 21% overall yield and with a longest linear sequence of just 11 steps from aldehyde 8. A key feature of the approach was the use of asymmetric permanganate-promoted oxidative cyclization to introduce four of the five required stereocenters in a single step. The use of robust and chemoselective methodology meant that the use of protecting groups could be avoided during the assembly of cis-solamin (1) from the three fragments 23, 6, and 4. The methodology was also applied to the synthesis of three further cis-solamin isomers 2, ent-1, and ent-2. Cytotoxicity and hemolytic properties of cis-solamin isomers and synthetic intermediates are reported. PMID- 15132545 TI - Synthesis of cyclic beta-amino acid esters from methionine, allylglycine, and serine. AB - Here we report a versatile ring-closing metathesis-based approach to 5-, 6-, and 7-membered cyclic beta-amino esters starting with simple and readily available building blocks-methionine, allylglycine, and serine-where the nature of the amino acid determines the size of the carbocyclic ring. PMID- 15132546 TI - Binding ability and assembly behavior of beta-cyclodextrin complexes with 2,2' dipyridine and 4,4'-dipyridine. AB - Two channel-type supramolecular aggregations 1 and 2 were prepared by the inclusion complex of beta-cyclodextrin with 2,2'-dipyridine and 4,4'-dipyridine, respectively, and their binding ability and assembly behavior were investigated comprehensively by X-ray crystallography, (1)H NMR, circular dichroism spectra, and microcalorimetric titration in solution and the solid state. The obtained results revealed that the hydrogen bonds and pi-pi stacking interactions are crucial factors for the formation of the molecular aggregations containing beta cyclodextrin and dipyridines. The disparity of nitrogen atom position in dipyridines leads not only to the distinct crystal system and space group, i.e., monoclinic system (C2) for 1 and triclinic system (P-1) for 2, but also different binding modes and thermodynamical parameters upon complexation of 2,2'-dipyridine and 4,4'-dipyridine with beta-cyclodextrin in aqueous solution. PMID- 15132547 TI - Mirror-image carbohydrates: synthesis of the unnatural enantiomer of a blood group trisaccharide. AB - Methyl D-glucoside and d-glucose pentaacetate are transformed, respectively, into methyl alpha-O-glucuronide 3 and hydroxymethyl beta-C-glucuronide 9, which undergo decarboxylative elimination efficiently to produce 4-deoxypentenoside 4 and L-glucal 10. These unsaturated pyranosides provide an expeditious entry into mirror-image oligosaccharides, as demonstrated in the synthesis of the unnatural enantiomer of the H-type II blood group determinant trisaccharide (D-Fuc-(alpha1- >2)-L-Gal-(beta1-->4)-L-GlcNAc-beta-OMe). This work illustrates that D-glucose, a common starting material in the synthesis of naturally occurring carbohydrates, can also be used to prepare their mirror-image analogues. PMID- 15132548 TI - An efficient way to 1,7-enynes and ethyl 8-yn-2-enoates from aldohexoses and to polyhydroxylated 1-vinylcyclohexenes. AB - In this paper, we report the synthesis of carbohydrate-derived 1,7-enynes and subsequent metathesis to yield polyhydroxylated 1-vinylcyclohexenes. For example, we converted D-glucose 2 to the (6,7)-dideoxy-D-gluco-hept-6-ene-pyranose 7, which led to the desired 1,7-enyne 16. The ring-closing metathesis of this 1,7 enyne 16 with the second generation Grubbs catalyst, under Mori's conditions, gave the corresponding polyhydroxylated 1-vinylcyclohexene 25 in 76% yield. The conversion of several aldohexoses into polyhydroxylated 1-vinylcyclohexenes was carried out with satisfying yields. We report also the synthesis of two carbohydrate-derived ethyl 8-yn-2-enoates from D-glucose derivatives. PMID- 15132549 TI - Ruthenium-catalyzed cycloaddition between propargylic alcohols and cyclic 1,3 dicarbonyl compounds via an allenylidene intermediate. AB - Thiolate-bridged diruthenium complexes such as [Cp*RuCl(mu(2)-SR)(2)RuCp*Cl] (Cp* = eta(5)-C(5)Me(5); R = Me, (n)Pr, (i)Pr) and [Cp*RuCl(mu(2) S(i)Pr)(2)RuCp*(OH(2))]OTf (OTf = OSO(2)CF(3)) promote the cycloaddition between propargylic alcohols and cyclic 1,3-dicarbonyl compounds to give either the corresponding 4,6,7,8-tetrahydrochromen-5-ones or 4H-cyclopenta[b]pyran-5-ones in high yields with complete regioselectivity. This catalytic cycloaddition provides a simple and one-pot synthetic protocol for a variety of substituted chromenones and cyclopenta[b]pyranones. PMID- 15132550 TI - Synthesis of azobenzenes from quinone acetals and arylhydrazines. AB - Direct reaction between quinone bisacetals and arylhydrazines gives azobenzenes. The presence of catalytic amounts of cerium ammonium nitrate strongly accelerates the reaction. When the bisacetal has a substituent at the 2,5-cyclohexadiene framework, only one regioisomer is formed. The method represents a simple, mild, and novel synthetic access to differently substituted azocompounds in high to excellent yield. PMID- 15132551 TI - Synthesis of 4,5-benzotropones by cyclization of 1,3-bis-silyl enol ethers with 1,2-dialdehydes. AB - The cyclization of 1,3-bis-silyl enol ethers or (2,4 dioxobutylidene)triphenylphosphoranes with phthalic dialdehyde allowed a convenient synthesis of a variety of 4,5-benzotropones. The hydrogenation of benzotropones afforded functionalized benzocycloheptanones which were transformed into tricyclic butenolides. PMID- 15132552 TI - Facile one-step synthesis of beta-alkoxy lactone via sequential lactonization and 1,4-addition of alkoxide group: total synthesis of all stereoisomers of dihydrokawain-5-ol. AB - We describe here a divergent total synthesis of all of the four stereoisomers of dihydrokawain-5-ol starting from alpha-D-glucose. The approach involves the use of Ando's modification of Horner-Wadsworth-Emmons homologation to give a alpha,beta-unsaturated ester. Subsequently, lactonization and 1,4-addition of OMe group in one step provided a delta-lactone, which was converted into the target compounds in two steps. PMID- 15132553 TI - Aerobic oxidation of benzyl alcohols catalyzed by aryl substituted N hydroxyphthalimides. Possible involvement of a charge-transfer complex. AB - A series of aryl-substituted N-hydroxyphthalimides (X-NHPIs) containing either electron-withdrawing groups (4-CH(3)OCO, 3-F) or electron-donating groups (4 CH(3), 4-CH(3)O, 3-CH(3)O, 3,6-(CH(3)O)(2)) have been used as catalysts in the aerobic oxidation of primary and secondary benzylic alcohols. The selective formation of aromatic aldehydes was observed in the oxidation of primary alcohols; aromatic ketones were the exclusive products in the oxidation of secondary alcohols. O-H bond dissociation enthalpies (BDEs) of X-NHPIs have been determined by using the EPR radical equilibration technique. BDEs increase with increasing the electron-withdrawing properties of the aryl substituent. Kinetic isotope effect studies and the increase of the substrate oxidation rate by increasing the electron-withdrawing power of the NHPI aryl substituent indicate a rate-determining benzylic hydrogen atom transfer (HAT) from the alcohol to the aryl-substituted phthalimide-N-oxyl radical (X-PINO). Besides enthalpic effects, polar effects also play a role in the HAT process, as shown by the negative rho values of the Hammett correlation with sigma(+) and by the decrease of the rho values (from -0.54 to -0.70) by increasing the electron-withdrawing properties of the NHPI aryl substituent. The relative reactivity of 3-CH(3)O-C(6)H(4)CH(2)OH and 3,4-(CH(3)O)(2)-C(6)H(3)CH(2)OH, which is higher than expected on the basis of the sigma(+) values, the small values of relative reactivity of primary vs secondary benzylic alcohols, and the decrease of the rho values by increasing the electron-withdrawing properties of the NHPI aryl substituent, suggest that the HAT process takes place inside a charge-transfer (CT) complex formed by the X PINO and the benzylic alcohol. PMID- 15132554 TI - Ab initio approach to understanding the stereoselectivity of reactions between hydroxyalkyl azides and ketones. AB - A new stereoselective version of the Schmidt reaction has been discovered by Aube and co-workers (J. Am. Chem. Soc. 2003, 125, 7914-7922). Quantum chemical calculations reported in this paper were carried out to examine the observed diastereoselectivities. The azide attack step is found to be reversible, but a thermodynamic preference for the equatorial attack product is observed. The final stereoselectivity of the reaction is determined by the axial/equatorial ratio of the chiral substituent in the resulting intermediate. In the case of 2-R hydroxypropyl azides, interesting axial/equatorial preferences are observed. In particular, the phenyl substituent shows a preference for the axial position resulting from a novel interaction with a N(2) cationic group. PMID- 15132555 TI - NiCl(2)(dppe)-catalyzed cross-coupling of aryl mesylates, arenesulfonates, and halides with arylboronic acids. AB - An investigation of the NiCl(2)(dppe)-, NiCl(2)(dppb)-, NiCl(2)(dppf)-, NiCl(2)(PCy(3))(2)-, and NiCl(2)(PPh(3))(2)-catalyzed cross-coupling of the previously unreported aryl mesylates, and of aryl arenesulfonates, chlorides, bromides, and iodides containing electron-withdrawing and electron-donating substituents with aryl boronic acids, in the absence of a reducing agent, is reported. NiCl(2)(dppe) was the only catalyst that exhibited high and solvent independent activity in the two solvents investigated, toluene and dioxane. NiCl(2)(dppe) with an excess of dppe, NiCl(2)(dppe)/dppe, was reactive in the cross-coupling of electron-poor aryl mesylates, tosylates, chlorides, bromides, and iodides. This catalyst was also efficient in the cross-coupling of aryl bromides and iodides containing electron-donating substituents. Most surprisingly, the replacement of the excess dppe from NiCl(2)(dppe)/dppe with excess PPh(3) generated NiCl(2)(dppe)/PPh(3), which was found to be reactive for the cross-coupling of both electron-rich and electron-poor aryl mesylates and chlorides. Therefore, the solvent-independent reactivity of NiCl(2)(dppe) provides an inexpensive and general nickel catalyst for the cross-coupling of aryl mesylates, tosylates, chlorides, bromides, and iodides with aryl boronic acids. PMID- 15132556 TI - Mechanistic studies on the Mukaiyama epoxidation. AB - A detailed mechanistic study on the Mukaiyama epoxidation of limonene with dioxygen as oxidant, bis(acetylacetonato)nickel(II) as catalyst, and an aldehyde as co-reagent is reported. All major products of the reaction have been quantitatively identified, both with isobutyraldehyde and 2-methylundecanal as co reacting aldehydes. Limonene epoxide is formed in good yield. The main products evolving from the aldehyde are carboxylic acid, CO(2), CO, and lower molecular weight ketone and alcohol (K + A). A mechanism is proposed in which an acylperoxy radical formed by the autoxidation of the aldehyde is the epoxidizing species. The observation of carbon dioxide and (K + A) in a 1:1 molar ratio supports this mechanism. CO(2) and (K + A) are formed in molar amounts of 50-60% with respect to the amount of epoxide produced, indicating that epoxidation takes place not only via acylperoxy radicals but also via a peracid route. Cyclohexene epoxidation was also investigated with a number of different metal complexes as catalysts. Cyclohexene is very sensitive for allylic oxidation, which provides information about the action of the catalyst, e.g., metals that form strongly oxidizing stable high-valence complexes are more likely to induce allylic oxidation. Color changes in the reaction mixture indicate the presence of such high-valence species. In the case of nickel, it was found that low-valence compounds predominate during the reaction, which is in line with the fact that this metal displays the highest selectivity for epoxide. A mechanism that accounts for the observations is presented. PMID- 15132557 TI - Aryl cations from aromatic halides. Photogeneration and reactivity of 4 hydroxy(methoxy)phenyl cation. AB - The photochemistry of 4-chlorophenol (1) and 4-chloroanisole (2) has been examined in a range of solvents and found to lead mainly to reductive dehalogenation, through a homolytic path in cyclohexane and a heterolytic path in alcohols. Heterolysis of 1 and 2 in methanol and 2,2,2-trifluoroethanol offers a convenient access to triplet 4-hydroxy- and 4-methoxyphenyl cations. These add to pi nucleophiles, viz., 2,3-dimethyl-2-butene, cyclohexene, and benzene, giving the arylated products in medium to good yields. Wagner-Meerwein hydride and alkyl migration are evidence for the cationic mechanism of the addition to alkenes. Arylation (with no rearrangement) was obtained to some extent also in nonprotic polar solvents such as MeCN and ethyl acetate, reasonably via an exciplex and with efficiency proportional to the nucleophilicity of the trap (2,3-dimethyl-2 butene > cyclohexene >> benzene). PMID- 15132558 TI - Allylation of alcohols and carboxylic acids with allyl acetate catalyzed by [Ir(cod)2](+)BF4- complex. AB - A facile method for the synthesis of allyl alkyl ethers from alcohols with allyl acetate was developed by the use of [Ir(cod)(2)](+)BF(4)(-) complex. For instance, the reaction of allyl acetate with n-octyl alcohol in the presence of a catalytic amount of [Ir(cod)(2)](+)BF(4)(-) complex afforded allyl octyl ether in quantitative yield. Allyl carboxylates were also prepared by the exchange reaction between carboxylic acids and allyl acetate in good yields. The [Ir(cod)(2)](+)BF(4)(-) complex catalyzed the reaction of alkyl and aromatic amines with allyl acetate to lead to the corresponding allylamines in fair to good yields. PMID- 15132559 TI - Addition of bifunctional organoboron reagents to strained alkenes. Carbon-carbon bond formation with Rh(I) catalysis in aqueous media. AB - Arylboronate esters bearing a pendant Michael-type acceptor olefin undergo transmetalation with a rhodium-based catalytic complex to generate a functionalized organorhodium intermediate that can cyclize onto strained olefins in good to excellent yields. The catalytic system involves the use of an electron rich, sterically bulky ligand to stabilize the organorhodium intermediate and reduce the incidence of protodeboronation in aqueous media. PMID- 15132560 TI - Total synthesis of (S)-(-)-(E)-15,16-dihydrominquartynoic acid: a highly potent anticancer agent. AB - The conjugated entriyne natural product, (S)-(E)-15,16-dihydrominquartynoic acid (1), is synthesized in five linear steps and 30% overall yield from the known aldehyde 11. The key step is a one-pot in situ desilylation/Cadiot-Chodkiewicz coupling reaction affording the entriyne unit. The bromoalkyne 6 with an omega carboxylic acid group was found to undergo a copper-catalyzed cross-coupling reaction producing the desired diyne intermediate 10, while the corresponding omega-ester bromoalkyne 14 failed to couple with triethylsilylacetylene under a variety of conditions. PMID- 15132561 TI - Hyperconjugation effect in substituted methyl boranes: an orbital deletion procedure analysis. AB - The hyperconjugation effect in the substituted methyl boranes, XCH(2)BH(2) (X = H, CH(3), NH(2), PH(2), OH, SH, F, Cl, Br), has been quantitatively evaluated by using the orbital deletion procedure (ODP), where the p(pi) orbital on boron is deactivated. Except for the case of X = NH(2), which forms a three-membered ring, the magnitude of the hyperconjugative stabilization in all other substituted methylborane ranges from 6.8 to 3.4 kcal/mol. Significant structural changes are observed, particularly the shortening of the central B-C bond distance and the reducing of the corresponding XCB and HCB bond angles. In general, the strength of the hyperconjugative interaction between the occupied sigma(C-X) bond and the vacant p(pi) orbital on boron is correlated to the electronegativity of X, and the competition between the donation ability of the sigma(C-X) and the sigma(C-H) bonds determines the preference of the staggered or eclipsed structure as the energy minimum state. When the donation abilities of the C-X and C-H bonds are comparable, other factors such as electron correlation and steric effect may play elaborate roles in the geometrical propensity of the most stable structures. PMID- 15132562 TI - Synthesis and conformational studies of a beta-turn mimetic incorporated in Leu enkephalin. AB - A beta-turn mimetic in which the four amino acids of a beta-turn have been replaced by a 10-membered ring has been designed, synthesized, and subjected to conformational studies. In the mimetic, the intramolecular CO(i)-HN(i)(+3) hydrogen bond that is often found in beta-turns has been replaced by an ethylene bridge. In addition, the amide bond between residues i and i + 1 was exchanged for a methylene ether isoster. Such a beta-turn mimetic, based on the first four residues of Leu-enkephalin (Tyr-Gly-Gly-Phe-Leu), was prepared in 15 steps. The synthesis relied on a beta-azido alcohol prepared in five steps from Cbz-Tyr(tBu) OH as a key, i-position building block. tert-Butyl bromoacetate, glycine, and a Phe-Leu dipetide were then used as building blocks for positions i + 1, i + 2, and i + 3, respectively. Conformational studies based on (1)H NMR data showed that the beta-turn mimetic was flexible, but that it resembled a type-II beta turn at low temperature. This low energy conformer closely resembled the structure determined for crystalline Leu-enkephalin. PMID- 15132563 TI - Fluorophore appended saccharide cyclophane: self-association, fluorescent properties, heterodimers with cyclodextrins, and cross-linking behavior with peanut agglutinin of dansyl-modified saccharide cyclophane. AB - A saccharide cyclophane bearing an environment-sensitive fluorophore (1) was prepared by introducing not only three branches with a terminal galactose residue but also one with a dansyl moiety into a tetraaza[6.1.6.1]paracyclophane skeleton. Self-association behavior of the dansyl-appended saccharide cyclophane was characterized in aqueous media by fluorescence spectroscopy and dynamic light scattering measurements. At least in the concentrations below 1.0 x 10(-5) M, saccharide cyclophane 1 existed in a monomeric state, whereas it tended to form self-aggregated complexes in the higher concentration. Solvent polarity dependency on the emission spectra of 1 was examined by fluorescence spectroscopy. With increasing dioxane contents in dioxane/water solvents, the fluorescence intensity originating from the dansyl moiety of 1 increased along with a concomitant blue shift of the fluorescence maximum (lambda(em)). In the monomeric state of 1 in water, the dansyl moiety of 1 was not fully included into its cyclophane cavity but partially exposed to the bulk aqueous phase. In the higher concentration ranges in an aggregate state, however, the dansyl group of 1 was located in the apolar cyclophane cavity whose microenvironment was equivalent to the polarity of 1-butanol evaluated on the basis of a correlation between lambda(em) and solvent polarity. This indicates an intermolecular inclusion of the dansyl moiety within the cyclophane. When cyclodextrin (CD) was mixed with 1, the dansyl group of 1 was bound to an internal cavity of CD such as gamma-CD, beta-CD, 6-O-alpha-glucosyl-beta-CD, and 6-O-alpha-maltosyl-beta-CD with binding constants of 7.5 x 10(2), 7.8 x 10(2), 7.7 x 10(2), and 6.0 x 10(2) M(-1), respectively. Such a supramolecular assembling of dansyl-modified cyclophane 1 and CDs caused changes of the fluorescence spectra as well as appearance of induced CD bands in aqueous media. Furthermore, saccharide cyclophane 1 was selectively bound to peanut agglutinin (PNA), galactoside-binding lectin, which was readily monitored by a visible turbidity of the solution due to a cross linking agglutination of these components, as well as by fluorescence spectroscopy. PMID- 15132564 TI - Zn(II)-induced ground-state pi-deconjugation and excited-state electron transfer in N,N-bis(2-pyridyl)amino-substituted arenes. AB - The synthesis and X-ray crystal structures of two N,N-bis(2-pyridyl)amino (dpa) substituted aromatic systems (Ar-dpa) 1 (Ar = 4,4'-disubstituted trans-stilbene) and 2 (Ar = 1,4-disubstituted benzene) and their ZnCl(2) complexes (1/ZnCl(2) and 2/ZnCl(2)) are reported. The fluoroionophoric behavior of 1-2 in response to Zn(II) in acetonitrile also has been investigated. In addition, compound 3DPA has been prepared and served as a pi-deconjugated model for 1DPA. The observed crystal structures for 1/ZnCl(2) and 2/ZnCl(2) could be divided into two distinct types, the planar and the twisted forms, depending on the aryl-dpa (C(ph)-NC(3)) dihedral angle. The twisted form is more favorable for these complexes unless the arene has a strong "push-pull" character. Nonetheless, the degree of pi conjugation between the N-pyridyl and the N-aryl group is reduced in both complex forms when compared with the free ligands. Such a Zn(II)-induced pi-deconjugation not only directly affects the internal charge transfer (ICT) fluorescence of the dpa-substituted stilbenes but also facilitates the occurrence of photoinduced electron transfer (PET) from the stilbene donor to the dpa/Zn(II) acceptor. The PET process is particularly important in accounting for the observed Zn(II) induced fluorescence quenching for 1DPA as well as 3DPA. PMID- 15132565 TI - Self-assembly of nanoscopic coordination cages using a flexible tripodal amide containing linker. AB - Flexible, nanoscopic 3D cages containing the amide functionality were prepared via coordination-driven self-assembly from palladium(II) based 90 degrees ditopic acceptor units and a tripod N,N',N''-tris(3-pyridyl)trimesic amide. Both cages were characterized by NMR ((31)P, (1)H) and electrospray ionization mass spectrometry. An MM2 force field simulation of one cage showed that the shape is likely pseudo trigonal bipyramidal with the diameter of the inner cavity of the cage about 1.9 nm. PMID- 15132566 TI - Total synthesis of petrobactin and its homologues as potential growth stimuli for Marinobacter hydrocarbonoclasticus, an oil-degrading bacteria. AB - A modular synthesis was developed to access petrobactin, a catechol-containing siderophore isolated from Marinobacter hydrocarbonoclasticus. A range of petrobactin homologues with differing dihydroxybenzamide motifs and in one case an increased number of carbons in the polyamine backbone were also synthesized. As such, these systems represent new isomeric probes to study iron transport properties in M. hydrocarbonoclasticus. The synthesis of petrobactin and its homologues and the first biological study of how these agents influence the growth of Mycobacterhydrocarbonoclasticus are reported. New synthetic methods were developed to overcome issues (imide formation) encountered in earlier syntheses. Both the (1)H and (13)C NMR of petrobactin were consistent with the recently revised structure showing that petrobactin in fact contains a 3,4 dihydroxybenzene motif rather than a 2,3-dihydroxybenzene motif. The preliminary biological studies suggested that using the native petrobactin 1b for M. hydrocarbonoclasticus-specific growth stimulation may be a poor strategy for oil spill cleanup. PMID- 15132567 TI - Photolysis of 3-(2-Formylphenyl)-3-chlorodiazirine in an Ar matrix at low temperature. AB - Photolysis of 3-(2-formylphenyl)-3-chlorodiazirine in an Ar matrix at 13 K was monitored by IR and UV-vis spectroscopies. Characterization of the products was carried out by comparing the spectra with the theoretical IR spectra obtained by using DFT, which suggested that anti,anti-2-formylphenyl(chloro)carbenes (a,a-2), ketenes (syn- and anti-3), chlorobenzocyclobutenone (4), and chloroisobenzofurane (5) were formed. This indicates that carbene 2 interacts with both hydrogen and oxygen in the formyl group. When the matrix containing a,a-2 was allowed to stand in the dark at 13 K, a gradual increase of the band due to anti-ketene 3 at the expense of 2 was observed. In contrast to its protio analogue, the deuterated carbene 2-d showed almost no changes either for the decay of 2-d or the growth of 3-d within the time range where the protio analogue appreciably decayed. These observations are interpreted in term of a quantum mechanical tunneling mechanism. PMID- 15132568 TI - Synthesis of cytochrome c oxidase models bearing a Tyr244 mimic. AB - A close structural analogue of the metal-free cytochrome c oxidase active site has been synthesized. This model has a proximal imidazole tail and three distal imidazole pickets attached to a porphyrin. One distal imidazole is cross-linked to a phenol, mimicking Tyr(244). The strategy behind the successful synthesis of this regioisomerically pure model involved discovering the best sequence to introduce the phenol-substituted imidazole and employing a fluorinated substituent. PMID- 15132569 TI - N-acylazinium salts: a new source of iminium ions for Ugi-type processes. AB - A multicomponent reaction involving the interaction of azines (quinolines, isoquinolines, and phenanthridine) with activating agents (chloroformates, acid halides, and sulfonyl halides), isocyanides, and water is described. The products of this process, alpha-carbamoylated-1,2-dihydroazines, are the result of the addition of the isocyanide partner to the N-acylazinium salt formed in situ. This represents a new source of iminium ion equivalents for Ugi-type reactions. PMID- 15132571 TI - Carbonylative ring expansion of aziridines to beta-lactams with rhodium-complexed dendrimers on a resin. AB - Rhodium-complexed dendrimers, supported on a resin, were evaluated as catalysts for the carbonylative ring expansion reactions of a variety of aziridines with carbon monoxide to give beta-lactams. The effects of reaction temperature, solvent, time, and pressure of carbon monoxide on this transformation were also investigated. The dendritic catalysts showed comparable activity to the homogeneous analogue. More importantly, this catalytic system can be easily recovered by simple filtration and recycled without significant loss of activity. PMID- 15132570 TI - Copper-mediated, palladium-catalyzed coupling of thiol esters with aliphatic organoboron reagents. AB - Thiol esters and B-alkyl-9-BBN derivatives couple in the presence of a copper(I) carboxylate mediator and a palladium catalyst. In contrast to copper-mediated, palladium-catalyzed cross-couplings of thioorganics with boronic acids, the current coupling reaction of 9-BBN derivatives is facilitated by the addition of a base such as Cs(2)CO(3). Under optimized conditions, a variety of thiol esters react with different B-alkyl-9-BBN derivatives giving ketones in moderate to excellent yields. PMID- 15132572 TI - Catalytic asymmetric carbalkoxyallylation of imines with the chiral bis-pi allylpalladium complex. AB - Carbethoxyallylstannane was employed along with the bis-pi-allylpalladium complex to achieve a useful conversion of prochiral imines to chiral 2-(2-aryl-2 aminoethyl)acrylates which are important building blocks for further asymmetric synthesis of a wide range of compounds. PMID- 15132573 TI - A general synthesis of iminosugars. AB - 1-Deoxynojirimycin, 1-deoxymannojirimycin, and 1-deoxygalactostatin have been synthesized by epoxidation of tri-O-acetyl-6-deoxyhex-5-enopyranosyl azides followed by methanolysis, deacetylation, and catalytic hydrogenation. 1,6 Dideoxygalactostatin was obtained by the reaction of 2,3,4-tri-O-acetyl-6-deoxy beta-L-arabino-hex-5-enopyranosyl azide with NIS in methanol followed by deacetylation and catalytic hydrogenation. The overall yields were 4.4-23.5% over seven to nine steps. PMID- 15132574 TI - Transition-metal-catalyzed aldehydic C-H activation by azodicarboxylates. AB - Rhodium acetate-catalyzed hydroacylation between aldehydes and an activated form of N=N bond was achieved under mild conditions to provide efficient access to a variety of hydrazino imides. Good selectivity for the aldehydic C-H activation relative to the ene-type reaction was observed with aldehydes having unsaturation both at terminal and internal positions. PMID- 15132575 TI - A convenient synthesis of a selective gelatinase inhibitor as an antimetastatic agent. AB - Compound 1, 2-(4-phenoxyphenylsulfonylmethyl)thiirane, is a potent and selective inhibitor for human gelatinases (J. Am. Chem. Soc. 2000, 122, 6799-6800), enzymes implicated in a number of diseases, including cancer. This compound is showing excellent promise in animal trials in a number of disease models. Large quantities of this compound were necessary for these studies. A convenient four step synthetic route for compound 1 is described herein. The synthesis is amenable to scale-up to tens of grams and gives an overall yield of 57% for this important compound. PMID- 15132576 TI - Conformational studies by dynamic NMR. 99. Experimental and computed determination of rotation barriers in the crystalline state: the case of naphthylphenylsulfoxide. AB - The (13)C NMR CP-MAS spectrum of 2-naphthylphenylsulfoxide in the solid state displays line broadening effects due to the restricted rotation about the Ph-S bond. Line shape simulation of the temperature-dependent traces allowed the corresponding barrier to be determined in the solids (14.7 kcal mol(-1)). By making use of the information obtained from single-crystal X-ray diffraction, this barrier could be satisfactorily reproduced by theoretical calculations (14.5 kcal mol(-1)) that take into account the correlated phenyl motion involving a large set of molecules in the crystalline state PMID- 15132578 TI - Application of functional ionic liquids possessing two adjacent acid sites for acetalization of aldehydes. AB - Several acid functional ionic liquids, in which cations possess two adjacent acid sites, were synthesized and used for the acetalization of aldehydes with good catalytic performance under mild reaction conditions. PMID- 15132577 TI - Pseudoamide-type pyrrolidine and pyrrolizidine glycomimetics and their inhibitory activities against glycosidases. AB - Coupling reaction of (2R,3R,4R,5R)-2,5-hydroxymethyl-3,4-dihydroxypyrrolidine (DMDP) with isothiocyanates afforded the corresponding thiourea adducts, which were transformed into isourea-type bicyclic oxapyrrolizidine glycomimetics by mercury(II) oxide-assisted intramolecular sulfur displacement. Cyclic carbamate and thiocarbamate analogues were also prepared by direct carbonylation or thiocarbonylation of DMDP. Evaluation of the glycosidase inhibitory properties demonstrated that remarkable specificities in enzyme inhibition can be achieved upon modifications on the pseudoaglyconic side chain and on the nature of the sp(2)-hybridized endocyclic ring nitrogen. PMID- 15132579 TI - Synthesis of sulfoxides by the hydrogen peroxide induced oxidation of sulfides catalyzed by iron tetrakis(pentafluorophenyl)porphyrin: scope and chemoselectivity. AB - The oxidation of sulfides with H(2)O(2) catalyzed by iron tetrakis(pentafluorophenyl)porphyrin in EtOH is an efficient and chemoselective process. With a catalyst concentration 0.03-0.09% of that of the substrate, sulfoxides are obtained with yields generally around 90-95% of isolated product. With vinyl and allyl sulfides, no epoxidation is observed. With a catalyst concentration between 0.09% and 0.25% of that of the substrate, sulfones are obtained in almost quantitative yield and with the same high chemoselectivity observed in the synthesis of sulfoxides. PMID- 15132580 TI - First practical protection of alpha-amino acids as N,N-benzyloxycarbamoyl derivatives. AB - The consecutive treatment of N-Cbz amino protected compounds with LiHMDS and CbzCl provides a practical method for the preparation of N,N-benzyloxycarbamoyl (N,N-di-Cbz) derivatives in good yield. When alpha-amino acids are used the protection occurs without racemization. The method is compatible with a wide range of other functional and protecting groups. The procedure is also valid for the synthesis of mixed N,N-carbamoyl derivatives. PMID- 15132581 TI - Selective oxidation of polyfunctional 2-amino-1,3-propanediol derivatives. AB - Oxidation of polyfunctional threo-(1S,2S)-2-amino-1,3-propanediol derivatives with a 717 anion-exchange resin-supported bromine has been investigated. The result showed that oxidized products were in close relationship with the substituents at nitrogen in the starting materials. Its primary and secondary amine derivatives were oxidized in the presence of Na(2)HPO(4) to give essentially a substituted chiral oxazoline or C(3)-O acylated product in high yield, while oxidation of its N,N-dimethyl derivative mainly gave a chiral N methyl oxidation-formylation product. This selective oxidation was first observed in 2-amino-1,3-propanediol chemistry. PMID- 15132582 TI - Organocatalysis of asymmetric epoxidation mediated by iminium salts under nonaqueous conditions. AB - The first nonaqueous conditions are described for catalytic asymmetric epoxidation mediated by iminium salt organocatalysts, providing ee values of up to 67%. PMID- 15132583 TI - Isotopic scrambling in Di-13C-labeled 2-butyl cation: evidence for a protonated cyclopropane intermediate. AB - The (13)C NMR spectrum of 2-butyl-1,2-(13)C(2) cation (1) is unchanged on heating the sample to -78 degrees C, indicating no isomerization to another isotopomer. In contrast, the spectrum of 2-butyl-2,3-(13)C(2) cation (2) shows rapid formation of all of the other isotopomers except 1. These results are consistent with a protonated cyclopropane intermediate in the rearrangement process. In this mechanism, either C(1) and C(2) or C(3) and C(4) interchange. Only the bond between C(2) and C(3) breaks. PMID- 15132585 TI - Molecular evolution on chiro-inositol dibenzoate using intramolecular acyl migration and selection by phenyl boronic acid. PMID- 15132586 TI - Image analysis as a quantitative screening test in combinatorial catalysis: discovery of an unexpected ruthenium-based catalyst for the Sonogashira reaction. PMID- 15132587 TI - Sidearm approach: a promising strategy for construction of bisoxazoline-based ligand library. PMID- 15132588 TI - Synthesis of tetrafurcated dendritic units on solid support. PMID- 15132589 TI - Array reactors for parallel synthesis. PMID- 15132590 TI - Combinatorial library of peptidotriazoles: identification of [1,2,3]-triazole inhibitors against a recombinant Leishmania mexicana cysteine protease. AB - A library consisting of about half of 800 000 possible peptidotriazoles on 450 000 beads was prepared by solid-phase peptide synthesis combined with a regiospecific copper(I)-catalyzed 1,3-dipolar cycloaddition between a resin-bound alkyne and a protected amino azide. The central [1,2,3]-triazole was flanked on each side by two randomized amino acids introduced in a combinatorial approach. Importantly, the formation of the triazole could be performed quantitatively in a randomized fashion. The library was screened on solid phase for inhibitory effect against a recombinant cysteine protease, Leishmania mexicana CPB2.8DeltaCTE and sorted by a high-throughput instrument, COPAS beadsorter (up to 200 000 beads/h). Forty-eight hits were analyzed by MALDI-TOF MS providing structural information about the protease specificity, and 23 peptidotriazoles were resynthesized and evaluated in solution, with the best inhibitor displaying a K(i) value of 76 nM. A one-pot procedure was used to convert Fmoc-amino azides into their corresponding Boc derivatives. The crucial influence of weak interactions with a spacer used for detection by MALDI-TOF MS on screening results was observed. PMID- 15132591 TI - A combinatorial approach for development of materials for optical sensing of gases. AB - We present a combinatorial approach for development of materials for use in optical gas sensors, with oxygen being used as an exemplary target gas. Combinatorial chemistry is shown to be a promising tool for speeding up the search for new sensor materials. The method is based on the use of various polymers, solvents, indicators, plasticizers, and other additives. Solutions of the respective materials are prepared in appropriate organic solvents, and a robotic station is programmed to mix the components. Spots of the sensing materials are deposited in the wells of glass substrates resembling microtiterplates. After drying off the solvent, the sensor spots are automatically analyzed in a test stand, where they are exposed to a carrier gas containing oxygen in various concentrations. Changes in the decay time of fluorescence of the indicator probes are measured and used (along with sensor response time) as a main criterion for sensor assessment. It is shown that the combinatorial approach can reduce the time and effort needed to establish libraries of sensor materials by a factor of at least 1000. We describe in detail the device for preparation of sensor libraries and for testing the respective materials. The potential of the system is demonstrated for the characterization of optical oxygen sensors. PMID- 15132592 TI - Fully automated polymer-assisted synthesis of 1,5-biaryl pyrazoles. AB - The polymer-assisted solution-phase (PASP) synthesis of a 192-member 2-D array of 1,5-biaryl pyrazoles 4[1-12,1-16] is reported. The synthesis was performed in a fully automated manner using a multiprobe top-filtration robot and incorporates a "catch and release" step to afford library compounds directly in high yield and purity. PMID- 15132593 TI - Tailoring ultraresins based on the cross-linking of polyethylene imines. Comparative investigation of the chemical composition, the swelling, the mobility, the chemical accessibility, and the performance in solid-phase synthesis of very high loaded resins. AB - Ultraresins have been prepared from polyethyleneimines and cross-linking molecules and have been provided with various degrees of cross-linking. The total nitrogen loading and the loading with secondary and with tertiary amines have been determined in all products. Nitrogen loadings of the novel resins were up to 15 mmol/g, reactive secondary amines up to 13.8 mmol/g. In addition to the exceptionally high loading, the novel resins displayed efficient swelling volumes in polar and nonpolar solvents. The mobility of resin-bound species as determined by EPR-spectroscopy, depending on the amount of cross-linker, indicated good flexibility and reactivity of this resin type. The novel, high-loaded resins have been investigated subsequently in solid-phase synthesis. The Rink amide linker and two different hydroxy linkers (hydroxyacetamide, HMPB) have been attached to the resin. Despite the high loadings, the secondary amines were easily accessible and could be functionalized exhaustively. Reactivity of the linker-coupled resins was found to be closely related to the resin composition. Increased resin cross linking led to reduced swelling, reduced mobility, and reduced reactivity in the synthesis of a medium-sized model peptide. As the result of the systematic investigation of structure-property relations in Ultraresins, a support material was identified that combined high reactivity and a mobility in the range of the extremely flexible Tentagel supports. In the optimized Ultraresin, >95% of all available secondary nitrogens could be coupled with Rink linker or with the small 2-hydroxyacetamide anchor, resulting in loadings from 2.7 to 6.8 mmol/g, respectively. A resin with an attached HMPB linker and spacer delivered analytically pure peptides in solid-phase synthesis, fully exploiting the exceptionally high loadings. PMID- 15132594 TI - gamma-Hydroxyalkynyl ketones as useful scaffolds for the preparation of combinatorial libraries of furans, isoxazoles and pyrazoles. AB - Using 5-hydroxy-hex-3-yne-2-one (9) as a model substrate, the utility of the gamma-hydroxyl alkynyl ketone scaffold for the preparation of combinatorial libraries of furans, isoxazoles, and pyrazoles is described. The addition of hydrazoic acid to the acetylenic ketone 9 forms in a single step both 3-azido-2,5 dimethyl furan (10) and alpha,5-dimethyl-3-isoxazolemethanol (11). Reaction of a mixture of (E)- and (Z)-bromoenediones (5 and 6), accessible from 3-bromo-2,5 dimethyl furan (17), with 1,1-dimethylhydrazine afforded a nearly quantitative yield of 1,3-dimethyl-5-acetylpyrazole (18). When the (E)- and (Z)-vinyl bromides 5 and 6 were reacted with sodium azide, 3-acetyl-5-methylisoxazole (7) formed via the intermediate (Z)-3-azido-3-hexene-2,5-dione (4), was the only product. PMID- 15132595 TI - Parallel solution-phase synthesis of (Z)-3-(arylamino)-2,3-dehydroalanine derivatives and solid-phase synthesis of fused pyrimidones. AB - N-Protected (Z)-3-(arylamino)-2,3-dehydroalanine esters 5 and 10 were prepared in one step from methyl (Z)-2-acylamino-3-(dimethylamino)prop-2-enoates 3 and 9 and anilines 4 employing a parallel solution-phase synthetic approach. In most cases, analytically pure products 5 and 10 were obtained. On the other hand, a three step parallel solid-phase synthesis of 2-acetylamino-4H-azino[1,2-x]pyrimidin-4 ones 15 via the polymer-bound methyl (Z)-2-acetylamino-3-(dimethylamino)prop-2 enoate (12) was also developed. PMID- 15132596 TI - Parallel synthesis and study of fluorous biphasic partition coefficients of 1H,1H,2H,2H-perfluoroalkylsilyl derivatives of triphenylphosphine: a statistical approach. AB - A library of fluorous, (1H,1H,2H,2H-perfluoroalkyl)silyl-substituted derivatives of triphenylphosphine, Ph(3-a)P[C(6)H(5-y)[SiMe(3 b)(CH(2)CH(2)C(x)F(2x+1))(b)](y)-pos](a) [a = 1-3; b = 1-3; x = 4, 6, 8, or 10; pos = 3, 4 (y = 1) or 3,5 (y = 2)], was prepared using parallel synthetic techniques. Upon variation of these four parameters, a total of 108 different fluorous phosphines can be synthesized. Using factorial design, 37 phosphines were selected and their partition coefficients in the typical fluorous biphasic solvent system PFMCH/toluene (PFMCH = perfluoromethylcyclohexane) determined. By fitting of the partition coefficient data to linear functions of the parameters a, b, and x, the partition coefficients of the remaining 71 fluorous phosphines, which were not prepared, could be predicted. Using this approach, some unexpected trends in the dependence of the partition coefficient on variations of the four parameters became clear, resulting in a better understanding of the optimum fluorous substitution pattern for obtaining the highest partition coefficient (P). In this way, the partition coefficient was increased by 2 orders of magnitude, i.e., from the initial value P = 7.8 for 1(3, 2, 6, C4) to P > 238 for 1(2, 3, 6, C3C5). Para- and 3,5-substituted phosphines showed irregular behavior in the sense that elongation or increase of the number of perfluoroalkyl tails did not necessarily lead to higher partition coefficients. Particularly high values were found for phosphines containing a total of 72 fluorinated carbon atoms on the meta position(s) of the aryl rings. Linear relationships were found between the predicted log P of 1(a, b, x, C4) and the experimentally determined log P values of fluorous diphosphines [CH(2)P[C(6)H(4)(SiMe(3 b)(CH(2)CH(2)C(6)F(13))(b))-4](2)](2) and monophosphines Ph(3 a)P(C(6)H(4)(CH(2)CH(2)C(6)F(13))-4)(a). One of the most fluorophilic phosphines, i.e., 1(3, 1, 8, C3C5), was applied and efficiently recycled in rhodium catalyzed, fluorous hydrosilylation of 1-hexene by HSiMe(2)Ph using PFMCH as the fluorous phase and the substrates as the organic phase. It was demonstrated that a higher partition coefficient of the ligand in PFMCH/toluene at 0 degrees C indeed resulted in less leaching of both the catalyst and the free ligand during phase separation. PMID- 15132597 TI - Development of an indole safety-catch linker using analytical constructs. AB - The development, evaluation, and application of a novel safety-catch linker for solid-phase synthesis based on an N-tosylindole is reported. The development of this linker using analytical constructs to aid analysis is discussed. PMID- 15132598 TI - Copper (II)-mediated arylation with aryl boronic acids for the N-derivatization of pyrazole libraries. AB - A N-derivatized 3-dimethylaminopropyloxypyrazole library was prepared using solution-phase parallel synthesis. The library was designed using physicochemical constraints designed to remove non-membrane-permeable molecules. Cupric acetate mediated N-arylation with aryl boronic acids proceeded regioselectively to form the N-2-substituted derivatives. The presence of the 3-dimethylaminopropyloxy group was found to completely control the regioselectivity of the arylation. Presence of a dimethylaminoethyloxy or dimethylaminobutyloxy group gave a lesser degree of regioselectivity. The scope of the method as applied to library synthesis is discussed. PMID- 15132599 TI - Fluorometric monitoring of organic reactions on solid phase. AB - The direct monitoring of reaction progress on solid supports by fluorescence spectroscopy is described. An immobilized fluorescent tracer molecule (dansyl chloride) is used to monitor the reaction on OH resins (Argopore Wang, PS Wang, and Argogel Wang), both in batch and in parallel chemistry. Fluorescence measurements were obtained directly on solid phase. The method demonstrated to be a valuable tool for the quantitative determination of resin-bound hydroxyl groups, to study reaction kinetics and for continuously monitoring the progress of the conversion of the hydroxyl resins into the chlorinated ones. The procedure proposed is highly sensitive compared to the traditional ones. The system can be extended to monitor a variety of reactions on solid supports, and in conjunction with a well-established technique such as flow analysis, basic studies on solid phase become possible. PMID- 15132600 TI - Optimization of antibacterial cyclic decapeptides. AB - A previously developed method for cyclic peptide synthesis was demonstrated to be able to provide convenient access to large combinatorial libraries of analogues, and this methodology was applied to the optimization of natural product cyclic decapeptides. Using this method, a 192-member library was designed and successfully constructed on the basis of the natural products tyrocidines, streptocidins, and loloatins to increase the therapeutic indices of these antibiotics. Library screening identified nine analogues whose therapeutic indices were increased by up to 90-fold in comparison to the natural products. Three of these analogues showed significant increase in antibacterial potency and concurrent drastic decrease in hemolytic activity. Since the natural products target the bacterial cell wall, the newly discovered analogues are promising leads for drug development against drug-resistant bacteria. PMID- 15132601 TI - Improved loading and cleavage methods for solid-phase synthesis using chlorotrityl resins: synthesis and testing of a library of 144 discrete chemicals as potential farnesyltransferase inhibitors. AB - The use of chlorotrityl resins for the immobilization of amines is sometimes deterred by the lengthy process of loading the reactants on the resins and product decomposition caused by the reactive chlorotrityl group in the presence of 1% TFA as a cleavage agent. Here, we report improved methods developed for selective and efficient loading of aminobenzoic acid derivatives on chlorotrityl resins and for cleavage of aniline-containing products from the resins without decomposition. These methods led to the synthesis of a library of 144 discrete chemicals as potential farnesyltransferase inhibitors (FTIs) using IRORI's radio frequency-encoded sorting technique and to the study of the applicability of the bivalence approach to the development of FTIs. PMID- 15132602 TI - Solid-Phase Synthesis of 2,4-diaminopyrimidines via Lewis acid-mediated aromatic nucleophilic substitution. AB - Primary amines were immobilized on (4-formyl-3,5 dimethoxyphenoxy)methylpolystyrene resin via reductive amination. Attachment of two different 4-chloro-2-methylthiopyrimidines, followed by sulfide oxidation, led to their corresponding sulfone intermediates. Aromatic nucleophilic substitution with various anilines or heteroaromatic amines in the presence of trimethyl aluminum afforded the desired 2,4-diaminopyrimidines after acidic cleavage from the resin. The synthetic methodology described herein was validated with the synthesis of a small 162-member library. PMID- 15132603 TI - Imaging reflection IR spectroscopy as a tool to achieve higher integration for high-throughput experimentation in catalysis research. AB - FTIR spectroscopy in reflection mode combined with a focal plane array (FPA) detector was employed for high-throughput screening of activity of catalysts in n pentane hydroisomerization. The reactor system was evaluated using reference catalysts Pt-MOR and gamma-alumina of known catalytic activity. By using the reflection setup, a higher degree of parallelization was possible, as compared to previous reports, in which transmission cells had been used. The 49-channel parallel reactor in combination with the FPA-IR optical setup was able to provide reliable information about the activity of different catalysts with relative data error of less than +/-20%. PMID- 15132604 TI - Combinatorial synthesis of substituted biaryls and heterocyclic arylamines. AB - In this paper, we report very general conditions that enable palladium-mediated coupling reactions on the solid support. A wide variety of biaryls and arylamines (including pyrimidines) have been synthesized using this protocol. The chemistry facilitates a combinatorial approach to the production of large numbers of medicinally relevant heterocyclic structures. PMID- 15132605 TI - Parallel approach for solution-phase synthesis of 2-quinoxalinol analogues and their inhibition of LPS-induced TNF-alpha release on mouse macrophages in vitro. AB - A parallel solution-phase synthesis of 2-quinoxalinol analogues is described. The key step-simultaneous reductions of m-Ar(NO2)2 to m-Ar(NH2)2 was investigated extensively. We obtained preliminary pharmacological activity of those analogues for the inhibition of LPS-induced TNF-alpha release on mouse macrophage in vitro. Two compounds revealed inhibitory activity, with IC50 values of 0.40 microM (7 amino-6-[(3-methoxypropyl)amino]-3-methyl-2-quinoxalinol) and 2.2 microM (7-amino 6-[(3-butoxypropyl)amino]-3-methyl-2-quinoxalinol), respectively. PMID- 15132607 TI - Preparation of Re(I)- and (99m)Tc(I)-metallocarboranes in water under weakly basic reaction conditions. AB - A new method for the preparation of Re- and (99m)Tc-metallocarboranes in water under mild reaction conditions was developed. Three nido-carborane ligands were reacted with [Re(CO)(3)Br(3)](2)(-) in the presence of aqueous potassium fluoride to give the corresponding eta(5)-Re(CO)(3)-carborane complexes. The use of KF as a base afforded the desired Re-metallocarboranes in good yields while avoiding the formation of Re clusters, which are byproducts commonly observed when reactions are carried out in the presence of strong aqueous bases. The reaction was also performed at the tracer level producing the first (99m)Tc-carborane complex, which was isolated in 80% radiochemical yield following a simple Sep-Pak purification process. The resulting organometallic complex was stable to cysteine and histidine challenges for more than 24 h. PMID- 15132606 TI - Parallel synthesis and antimalarial screening of a 4-aminoquinoline library. AB - Due to growing problems with drug resistance, there is an outstanding need for new, cost-effective drugs for the treatment of malaria. The 4-aminoquinolines have provided a number of useful antimalarials, and Plasmodium falciparum, the causative organism for the most deadly form of human malaria, is generally slow to develop resistance to these drugs. Therefore, diverse screening libraries of quinolines continue to be useful for antimalarial drug discovery. We report herein the development of an efficient method for producing libraries of 4 aminoquinolines variant in the side chain portion of the molecule. The effects of these substitutions were evaluated by screening this library for activity against P. falciparum, revealing four potent compounds active against drug-resistant strains. PMID- 15132608 TI - The observation of the first vibrational overtone of dihydrogen in the luminescence of zeolites at low temperatures. AB - Dehydrated zeolites NaA, NaY, and barium-exchanged NaY luminesce when irradiated with the 1064 nm laser light of an FT-Raman instrument. When hydrogen is adsorbed in the zeolite, the luminescence is altered in several ways. Most remarkable is the appearance of "absorptions" in the positions of the first vibrational overtones of H(2) and HD. Although these features are in the expected positions with reasonable band profiles for overtone absorptions, the large extinction of the luminescent intensity requires a more efficient mechanism than simple reabsorption of emitted photons. In addition to the appearance of holes in the luminescence spectrum, other luminescent features are substantially quenched by the presence of hydrogen or, in one case, augmented. PMID- 15132609 TI - Using model complexes to augment and advance metalloproteinase inhibitor design. AB - The tetrahedral zinc complex [(Tp(Ph,Me))ZnOH] (Tp(Ph,Me) = hydrotris(3,5 phenylmethylpyrazolyl)borate) was combined with 2-thenylmercaptan, ethyl 4,4,4 trifluoroacetoacetate, salicylic acid, salicylamide, thiosalicylic acid, thiosalicylamide, methyl salicylate, methyl thiosalicyliate, and 2 hydroxyacetophenone to form the corresponding [(Tp(Ph,Me))Zn(ZBG)] complexes (ZBG = zinc-binding group). X-ray crystal structures of these complexes were obtained to determine the mode of binding for each ZBG, several of which had been previously studied with SAR by NMR (structure-activity relationship by nuclear magnetic resonance) as potential ligands for use in matrix metalloproteinase inhibitors. The [(Tp(Ph,Me))Zn(ZBG)] complexes show that hydrogen bonding and donor atom acidity have a pronounced effect on the mode of binding for this series of ligands. The results of these studies give valuable insight into how ligand protonation state and intramolecular hydrogen bonds can influence the coordination mode of metal-binding proteinase inhibitors. The findings here suggest that model-based approaches can be used to augment drug discovery methods applied to metalloproteins and can aid second-generation drug design. PMID- 15132610 TI - The ammine, thiosulfato, and mixed ammine/thiosulfato complexes of silver(I) and gold(I). AB - The M(I)-NH(3), M(I)-S(2)O(3)(2)(-), and M(I)-S(2)O(3)(2)(-)-NH(3) systems (M = Ag, Au) were studied at 25 degrees C and at I = 0.1 M (NaClO(4)) using a variety of analytical techniques. For the Ag(I)-NH(3)-S(2)O(3)(2)(-) system, AgS(2)O(3)NH(3)(-) was detected with formation constant log beta(111) (for the reaction Ag(+) + S(2)O(3)(2)(-) + NH(3) <--> AgS(2)O(3)NH(3)(-)) of 11.2, 10.4, and 10.8 on the basis of silver potentiometry, UV-vis spectrophotometry, and hydrodynamic voltammetry, respectively. Also, the values of log beta(101)(AgNH(3)(+)), log beta(102)(Ag(NH(3))(2)(+)), log beta(110)(AgS(2)O(3)( )), and log beta(120)(Ag(S(2)O(3))(2)(3)(-)), obtained from silver potentiometry, were 3.59, 7.0, 8.97, 13.1, respectively. In the case of the ammine complexes, the log beta(101)(AgNH(3)(+)) and log beta(102)(Ag(NH(3))(2)(+)) values were found to be 3.5 and 7.1, respectively, from the UV-vis spectrophotometric experiments. The mixed species AuS(2)O(3)NH(3)(-) was detected in UV-vis spectrophotometric, hydrodynamic voltammetric, and potentiometric experiments with the stepwise formation constants (log K(111)) of -4.0, -3.5, -3.8, respectively, for the reaction Au(S(2)O(3))(2)(3)(-) + NH(3) <--> AuS(2)O(3)NH(3)(-) + S(2)O(3)(2)(-). At higher [NH(3)]/[S(2)O(3)(2)(-)] ratios (>10(5)), the formation of Au(NH(3))(2)(+) was also detected in spectrophotometric and potentiometric experiments with stepwise formation constants (log K(102)) of -5.4 and -5.3, respectively, according to the reaction AuS(2)O(3)NH(3)(-) + NH(3) <--> Au(NH(3))(2)(+) + S(2)O(3)(2)(-). PMID- 15132611 TI - Dyads containing iridium(III) bis-terpyridine as photoactive center: synthesis and electron transfer study. AB - A series of Ir(III)-D dyads based on an iridium(III) bis-terpyridine complex as a photoactive center and tertiary amines as donor groups, as well as their individual components, have been designed to generate photoinduced charge separation. Depending on the donor group, a modular approach or a "chemistry-on the-complex" approach has been used to prepare three different Ir(III)-D dyads. A detailed photophysical study has been performed on one Ir(III)-D dyad in which a triarylamine is linked to the iridium bis-terpyridine complex with an amido phenyl group used as a spacer. In acetonitrile at room temperature, steady-state and time-resolved methods gave evidence of a photoinduced charge-separated state Ir(-)-D(+) with a lifetime of 70 ps. This relatively short lifetime could be due to the close proximity between the negative charge, likely localized in the bridging terpyridine, and the oxidized donor group. PMID- 15132613 TI - Structural and thermodynamic properties of group 13 imidometallanes and their heavier analogues. AB - Systematic theoretical studies of the [XMYH](n) inorganic rings and clusters (M = Al, Ga, In; Y = N, P, As; X = H, F, Cl, Br, I; n = 1-6) have been carried out using hybrid Hartree-Fock density functional theory. A consistent set of the structural and thermodynamic properties has been obtained. The stability of the MY bond decreases in the order Al > Ga >or= In; N >> P > As. Terminal groups X have a minor influence on the subsequent elimination enthalpies of the clusters. In the case of X = H, hydrogen elimination makes formation of the [HMYH](6) oligomers from MH(3) and YH(3) thermodynamically favorable; while in the case of halide substituents, formation of [XMYH](6) is thermodynamically unfavorable, except for the system with the strongest MY bond (AlN). Substitution of the acidic hydrogen by X is favorable energetically for all [HMYH](6) clusters, but is complicated by the processes of cluster destruction to form the [X(2)MYH(2)](2) dimers. The high stability of the [HMNH](6) clusters makes them attractive single-source precursors for the production of 13-15 composites. PMID- 15132612 TI - A structural and Mossbauer study of complexes with Fe(2)(micro-O(H))(2) cores: stepwise oxidation from Fe(II)(micro-OH)(2)Fe(II) through Fe(II)(micro OH)(2)Fe(III) to Fe(III)(micro-O)(micro-OH)Fe(III). AB - Dinuclear non-heme iron clusters containing oxo, hydroxo, or carboxylato bridges are found in a number of enzymes involved in O(2) metabolism such as methane monooxygenase, ribonucleotide reductase, and fatty acid desaturases. Efforts to model structural and/or functional features of the protein-bound clusters have prompted the preparation and study of complexes that contain Fe(micro-O(H))(2)Fe cores. Here we report the structures and spectroscopic properties of a family of diiron complexes with the same tetradentate N4 ligand in one ligand topology, namely [(alpha-BPMCN)(2)Fe(II)(2)(micro-OH)(2)](CF(3)SO(3))(2) (1), [(alpha BPMCN)(2)Fe(II)Fe(III)(micro-OH)(2)](CF(3)SO(3))(3) (2), and [(alpha BPMCN)(2)Fe(III)(2)(micro-O)(micro-OH)](CF(3)SO(3))(3) (3) (BPMCN = N,N'-dimethyl N,N'-bis(2-pyridylmethyl)-trans-1,2-diaminocyclohexane). Stepwise one-electron oxidations of 1 to 2 and then to 3 demonstrate the versatility of the Fe(micro O(H))(2)Fe diamond core to support a number of oxidation states with little structural rearrangement. Insight into the electronic structure of 1, 2', and 3 has been obtained from a detailed Mossbauer investigation (2' differs from 2 in having a different complement of counterions). Mixed-valence complex 2' is ferromagnetically coupled, with J = -15 +/- 5 cm(-)(1) (H = JS(1).S(2)). For the S = (9)/(2) ground multiplet we have determined the zero-field splitting parameter, D(9/2) = -1.5 +/- 0.1 cm(-)(1), and the hyperfine parameters of the ferric and ferrous sites. For T < 12 K, the S = (9)/(2) multiplet has uncommon relaxation behavior. Thus, M(S) = -(9)/(2) <--> M(S) = +(9)/(2) ground state transition is slow while deltaM(S) = +/-1 transitions between equally signed M(S) levels are fast on the time scale of Mossbauer spectroscopy. Below 100 K, complex 2' is trapped in the Fe(1)(III)Fe(2)(II) ground state; above this temperature, it exhibits thermally assisted electron hopping into the state Fe(1)(II)Fe(2)(III). The temperature dependence of the isomer shifts was corrected for second-order Doppler shift, obtained from the study of diferrous 1. The resultant true shifts were analyzed in a two-state hopping model. The diferric complex 3 is antiferromagnetically coupled with J = 90 +/- 15 cm(-)(1), estimated from a variable-temperature Mossbauer analysis. PMID- 15132614 TI - Synthesis, structure, and dynamics of nickelacarboranes incorporating the [nido 7,9-C(2)B(9)H(11)](2)(-) ligand. AB - The nickelacarboranes [NEt(4)][2-(eta(3)-C(3)H(4)R)-closo-2,1,7-NiC(2)B(9)H(11)] (R = H (1a), Ph (1b)) have been synthesized via reaction between [Na](2)[nido-7,9 C(2)B(9)H(11)] and [Ni(2)(micro-Br)(2)(eta(3)-C(3)H(4)R)(2)] in THF (THF = tetrahydrofuran), followed by addition of [NEt(4)]Cl. Protonation of 1a in the presence of a donor ligand L affords the complexes [2,2-L(2)-closo-2,1,7 NiC(2)B(9)H(11)] (L = CO (2), CNBu(t) (3)). Addition of PEt(3) (1 equiv) to 2 produces quantitative conversion to [2-CO-2-PEt(3)-closo-2,1,7-NiC(2)B(9)H(11)], 4. Species 2-4 exhibit in solution hindered rotation of the NiL(2) fragment with respect to the eta(5)-C(2)B(9) cage unit. Protonation of 1a in the presence of a diene affords the neutral complexes [2-(eta(2):eta(2)-diene)-closo-2,1,7 NiC(2)B(9)H(11)] (diene = C(5)Me(5)H (5), dcp (6), cod (7), nbd (8), chd (9), and cot (10a); dcp = dicyclopentadiene, cod = 1,5-cyclooctadiene, nbd = norbornadiene, chd = 1,3-cyclohexadiene, and cot = cyclooctatetraene). Variable temperature (1)H NMR experiments show that the [Ni(diene)] fragments are freely rotating even at 193 K. A small quantity of the di-cage species [2,2'-micro (1,2:5,6-eta-3,4:7,8-eta-cot)-(closo-2,1,7-NiC(2)B(9)H(11))(2)] (10b) is formed as a coproduct in the synthesis of 10a. This species can be rationally synthesized by protonation of 1a and subsequent addition of 10a. PMID- 15132615 TI - Proton transfer to nickel-thiolate complexes. 1. Protonation of [Ni(SC(6)H(4)R 4)(2)(Ph(2)PCH(2)CH(2)PPh(2))] (R = Me, MeO, H, Cl, or NO(2)). AB - The kinetics of the equilibrium reaction between [Ni(SC(6)H(4)R-4)(2)(dppe)] (R= MeO, Me, H, Cl, or NO(2); dppe = Ph(2)PCH(2)CH(2)PPh(2)) and mixtures of [lutH](+) and lut (lut = 2,6-dimethylpyridine) in MeCN to form [Ni(SHC(6)H(4)R 4)(SC(6)H(4)R-4)(dppe)](+) have been studied using stopped-flow spectrophotometry. The kinetics for the reactions with R = MeO, Me, H, or Cl are consistent with a single-step equilibrium reaction. Investigation of the temperature dependence of the reactions shows that DeltaG = 13.6 +/- 0.3 kcal mol(-)(1) for all the derivatives but the values of DeltaH and DeltaS vary with R (R = MeO, DeltaH() = 8.5 kcal mol(-)(1), DeltaS = -16 cal K(-)(1) mol(-)(1); R = Me, DeltaH() = 10.8 kcal mol(-)(1), DeltaS = -9.5 cal K(-)(1) mol(-)(1); R = Cl, DeltaH = 23.7 kcal mol(-)(1), DeltaS = +33 cal K(-)(1) mol(-)(1)). With [Ni(SC(6)H(4)NO(2)-4)(2)(dppe)] a more complicated rate law is observed consistent with a mechanism in which initial hydrogen-bonding of [lutH](+) to the complex precedes intramolecular proton transfer. It seems likely that all the derivatives operate by this mechanism, but only with R = NO(2) (the most electron withdrawing substituent) does the intramolecular proton transfer step become sufficiently slow to result in the change in kinetics. Studies with [lutD](+) show that the rates of proton transfer to [Ni(SC(6)H(4)R-4)(2)(dppe)] (R = Me or Cl) are associated with negligible kinetic isotope effect. The possible reasons for this are discussed. The rates of proton transfer to [Ni(SC(6)H(4)R 4)(2)(dppe)] vary with the 4-R-substituent, and the Hammett plot is markedly nonlinear. This unusual behavior is attributable to the electronic influence of R which affects the electron density at the sulfur. PMID- 15132616 TI - Proton transfer to nickel-thiolate complexes. 2. Rate-limiting intramolecular proton transfer in the reactions of [Ni(SC(6)H(4)R 4)(PhP[CH(2)CH(2)PPh(2)](2))](+) (R = NO(2), Cl, H, Me, or MeO). AB - The protonation of [Ni(SC(6)H(4)R-4)(triphos)](+) (triphos = PhP[CH(2)CH(2)PPh(2)](2); R = NO(2), Cl, H, Me, or MeO) by [lutH](+) (lut = 2,6 dimethylpyridine) to form [Ni(S(H)C(6)H(4)R-4)(triphos)](2+) is an equilibrium reaction in MeCN. Kinetic studies, using stopped-flow spectrophotometry, reveal that the reactions occur by a two-step mechanism. Initially, [lutH](+) rapidly binds to the complex (K(2)(R)) in an interaction which probably involves hydrogen bonding of the acid to the sulfur. Subsequent intramolecular proton transfer from [lutH](+) to sulfur (k(3)(R)) is slow because of both electronic and steric factors. The X-ray crystal structures of [Ni(SC(6)H(4)R-4)(triphos)](+) (R = NO(2), H, Me, or MeO) show that all are best described as square-planar complexes, with the phenyl substituents of the triphos ligand presenting an appreciable barrier to the approach of the sterically demanding [lutH](+) to the sulfur. The kinetic characteristics of the intramolecular proton transfer from [lutH](+) to sulfur have been investigated. The rate of intramolecular proton transfer exhibits a nonlinear dependence on Hammett sigma(+), with both electron releasing and electron-withdrawing 4-R-substituents on the coordinated thiolate facilitating the rate of proton transfer (NO(2) > Cl > H > Me < MeO). The rate constants for intramolecular proton transfer correlate well with the calculated electron density of the sulfur. The temperature dependence of the rate of the intramolecular proton transfer reactions shows that deltaH() is small but increases as the 4-R-substituent becomes more electron-withdrawing [deltaH = 4.1 (MeO), 6.9 (Me), 11.4 kcal mol(-)(1) (NO(2))], while DeltaS() becomes progressively less negative [deltaS = -50.1 (MeO), -41.2 (Me), -16.4 (NO(2)) cal K(-)(1) mol(-)(1)]. Studies with [lutD](+) show that the rate of intramolecular proton transfer varies with the 4-R-substituent [(k(3)(NO)2)(H)/(k(3)(NO)2)(D) = 0.39; (k(3)(Cl))(H)/(k(3)(Cl))(D) = 0.88; (k(3)(Me))(H)/(k(3)(Me))(D) = 1.3; (k(3)(MeO))(H)/(k(3)(MeO))(D) = 1.2]. PMID- 15132617 TI - Complex formation of vanadium(IV) with 1,3,5-triamino-1,3,5-trideoxy-cis-inositol and related ligands. AB - The complex formation of vanadium(IV) with 1,3,5-triamino-1,3,5-trideoxy-cis inositol (taci) and 1,3,5-trideoxy-1,3,5-tris(dimethylamino)-cis-inositol (tdci) was studied in aqueous solution and in the solid state. The formation constants of [V(IV)O(taci)](2+), [V(IV)O(tdci)](2+), and [V(IV)(tdci)(2)](4+) and of the deprotonation product [V(IV)(tdci)(2)H(-)(1)](3+) were determined (25 degrees C, 0.1 M KNO(3)). Cyclic voltammetry measurements established a reversible one electron transfer for the [V(IV)(tdci)(2)H(-)(m)]((4)(-)(m))/[V(III)(tdci)(2)H( )(n)]((3)(-)(n)) couple (0 pi(NC)/pi(PPh(2)). PMID- 15132619 TI - Unexpected reactivity of PdCl(2) and PtCl(2) complexes of the unsaturated diphosphine o-Me(2)TTF(PPh(2))(2) toward chloride abstraction with thallium triflate. AB - Attempted thallium triflate abstraction of chloride anions from the MCl(2) complexes of the unsaturated chelating diphosphines o-dimethyl bis(diphenylphosphino)tetrathiafulvalene (P2) (M = Pd, Pt) and cis-1,2 bis(diphenylphosphino)ethylene (dppen) (M = Pd) affords, surprisingly, a Tl(OTf) adduct in the case of (P2)PdCl(2) and (P2)PtCl(2), with no chloride abstraction, and a dicationic bis(palladium) bis(triflate) salt in the case of (cis dppen)PdCl(2), in which only one Cl anion was removed. The crystal structures of these complexes were determined by X-ray analysis, which established the formulations (P2)MCl(2).Tl(OTf) (M = Pd, Pt) and [(dppen)PdCl](2)(OTf)(2), respectively. These compounds can be seen as possible intermediates in the general chloride abstraction process between (P-P)MCl(2) (M = Pd, Pt) and thallium triflate. PMID- 15132620 TI - Assignment of the electronic spectra of [Mo(CN)(8)](4)(-) and [W(CN)(8)](4)(-) by Ab initio calculations. AB - CASPT2 calculations are performed on the dodecahedral and square antiprismatic isomers of the [Mo(CN)(8)](4)(-) and [W(CN)(8)](4)(-) complexes. The high-energy experimental bands above 40000 cm(-)(1) are assigned to MLCT transitions. The experimental observed trend of the extinction coefficients for the molybdenum and tungsten complex is reproduced by our CASSCF oscillator strengths. All bands below 40000 cm(-)(1) can be ascribed to ligand-field transitions, although small contributions from forbidden MLCT transitions cannot be excluded. In order to account for all experimental bands in the electronic spectrum of these octacyanocomplexes, a dynamic equilibrium in solution between the two isomeric forms must be hypothesized. Spin-orbit coupling effects are found to be more important for the square antiprismatic isomers; in particular, large singlet triplet mixings are calculated for this isomer of [W(CN)(8)](4)(-). Ligand-field and Racah parameters as well as spin-orbit coupling constants are determined on the basis of the calculated transition energies. The obtained values for these parameters support the recently proposed model for exchange interactions in magnetic clusters and networks containing pentavalent octocyanometalates of molybdenum and tungsten. PMID- 15132621 TI - Electron delocalization in nickel metallic wires: a DFT investigation of Ni(3)(dpa)(4)Cl(2) and [Ni(3)(dpa)(4)](3+) (dpa = dipyridylamide) and extension to higher nuclearity chains. AB - The electronic structure of Ni(3)(dpa)(4)Cl(2) (1) has been investigated within the framework of the density functional theory (DFT), using two types of exchange correlation functionals and various basis sets. The "broken-symmetry" approach proposed by Noodleman for the characterization of electronic states displaying an antiferromagnetic coupling has been applied to 1. All calculations lead to the conclusion that the ground state results from an antiferromagnetic coupling between the terminal Ni atoms, both displaying a high-spin electronic configuration. The central Ni atom is in a low-spin configuration, but is involved in a superexchange interaction connecting the two magnetic centers. These results are in agreement with the assignments recently proposed by the group of F. A. Cotton on the basis of magnetic measurements. It is shown that the ground state electronic configuration calculated for 1 provides the trinickel framework with some delocalized sigma bonding character. The observed geometry of 1 is accurately reproduced by the broken-symmetry solution. The doublet ground state assigned to the oxidized species [Ni(3)(dpa)(4)](3+) (2) and the dramatic contraction of the coordination sphere of the terminal metals observed upon oxidation are also confirmed by the calculations. However, the formal Ni-Ni bond order is not expected to increase in the oxidized species. The contraction of the Ni-Ni distance in 2 is shown to result in part from the vanishing of the important trans influence originating in the axial ligands, and for the rest from a more efficient shielding of the metal nuclear charge along the Ni-Ni-Ni axis. The conclusions deduced from the analysis of the bonding in 1 and 2 can be extended to their homologues with higher nuclearity. More specifically, it is predicted that the single occupancy of the most antibonding sigma orbital, extending over the whole metal framework, will provide the (Ni(p))(2)(p)(/(2)(p)(+1)+) chains with some delocalized bonding character and, possibly, with electrical conduction properties. PMID- 15132622 TI - Synthesis and characterization of a new three-dimensional lanthanide carboxyphosphonate: Ln(4)(H(2)O)(7)[O(2)C-C(5)H(10)N-CH(2)(-)PO(3)](4)(H(2)O)(5). AB - The first open-framework lanthanide carboxyphosphonate has been obtained under hydrothermal conditions. The three-dimensional structure of MIL-84(Pr) (MIL = Material Institut Lavoisier) or Pr(4)(H(2)O)(7)[O(2)C-C(5)H(10)N-CH(2)( )PO(3)](4)(H(2)O)(5) has been solved from X-ray diffraction single-crystal data (a = 23.481(1) A, b = 10.159(1) A, c = 23.006(1) A, beta = 105.63(1) degrees, V = 5284.6(6) A(3), space group Cc (No. 9)). Its framework is built up from chains of edged-sharing eight or nine-coordinated monocapped square antiprism polyhedra and carboxyphosphonate anions, creating a three-dimensional structure with small pores filled with water molecules. The thermal behavior of MIL-84(Pr) has been investigated using TGA and X-ray thermodiffractometry and indicates that MIL 84(Pr) is stable up to 523 K with a reversible hydration-dehydration process. The optical study of its yttrium analogue doped at 3.4% with europium (MIL-84(Y,Eu)) reveals a significant red-orange emission under UV radiation. PMID- 15132623 TI - Defect-induced acceleration of a solid-state chemical reaction in zinc alkoxide single crystals. AB - A family of dinuclear (salicylaldimato)zinc(micro(2)-alkoxide) complexes show unusual behavior, decomposing to olefins and the corresponding micro(2)-hydroxide complexes under very mild conditions in the solid state. The reaction in homogeneous solution is, on the other hand, not observable, even at elevated temperatures over extended periods of time. The reaction in the solid state occurs preferentially on one crystallographic face and displays kinetics characteristic of polymorphic transformations in single crystals. PMID- 15132624 TI - Distorted equatorial coordination environments and weakening of U=O bonds in uranyl complexes containing NCN and NPN ligands. AB - Treatment of [UO(2)Cl(2)(thf)(3)] in thf with 2 equiv of Na[PhC(NSiMe(3))(2)] (Na[NCN]) or Na[Ph(2)P(NSiMe(3))(2)] (Na[NPN]) gives uranyl complex [UO(2)(NCN)(2)(thf)] (1) or [UO(2)(NPN)(2)] (3), respectively. Each complex is a rare example of out-of-plane equatorial nitrogen ligand coordination; the latter contains a significantly bent O=U=O unit and represents the first example of a uranyl ion within a quadrilateral-faced monocapped trigonal prismatic geometry. Removal of the thf in 1 gives [UO(2)(NCN)(2)] (2) with in-plane N donor ligands. Addition of 3 equiv of Na[NCN] gives the tris complex [Na(thf)(2)PhCN][[UO(2)(NCN)(3)] (4.PhCN) with elongation and weakening of one U=O bond through coordination to Na(+). Hydrolysis of 4 provides the oxo-bridged dimer [Na(thf)UO(2)(NCN)(2)](2)(micro(2)-O) (6), a complex with the lowest reported O=U=O symmetrical stretching frequency (nu(1) = 757 cm(-)(1)) for a dinuclear uranyl complex. The anion in complex 4 is unstable in solution but can be stabilized by the introduction of 18-crown-6 to give [Na(18-crown 6)][UO(2)(NCN)(3)] (5). The structures of 1-4 and 6 have been determined by crystallography, and all except 2 show significant deviations of the N ligand atoms from the equatorial plane, driven by the steric bulk of the NCN and NPN ligands. Despite the unusual geometries, these distortions in structure do not appear to have any direct effect on the bonding and electronic structure of the uranyl ion. The main influences toward lowering the U=O bond stretching frequency (nu(1)) are the donating ability of the equatorial ligands, overall charge of the complex, and U=O.Na-type interactions. The intense orange/red colors of these compounds are because of low-energy ligand-to-metal charge-transfer electronic transitions. PMID- 15132625 TI - Carbon-sulfur bond cleavage in Bis(N-alkyldithiocarbamato)cadmium(II) complexes: heterolytic desulfurization coupled to topochemical proton transfer. AB - Three bis(N-alkyldithiocarbamato)cadmium(II) complexes [Cd(S(2)CNHR)(2)] (1, R = n-C(3)H(7); 2, R = n-C(5)H(11); 3, n-C(12)H(25)) were prepared by metathesis of the corresponding lithium salt, Li[S(2)CNHR], with cadmium chloride. The crystal structures of 2 and 3 consist of planar molecular units of [Cd(S(2)CNHR)(2)] connected by intermolecular Cd.S interactions to give a one-dimensional chain. The chains are connected by a network of intermolecular N-H.S hydrogen bonds between the dithiocarbamato nitrogen atom and bridging sulfur atoms in neighboring chains. In solution, the (113)Cd NMR spectrum of 2 is dependent on concentration and temperature, indicative of a dimerization equilibrium mediated by similar Cd.S intermolecular bridging interactions. In the solid state, thermal gravimetric analyses show that all three complexes decompose smoothly via a heterolytic C-S bond cleavage reaction to give the corresponding alkyl isothiocyanate and cadmium sulfide as the primary products, with the formation of primary amine and CS(2) as coproducts. These products can result only from the net transfer of protons between N-alkyldithiocarbamato ligands in the solid state. Thus, the C-S bond cleavage reaction is interpreted in terms of the topochemical arrangement of molecular units in the crystalline state, which provides a pathway for proton transfer between ligands via N-H.S hydrogen bonds. Decomposition was also initiated by addition of a tertiary amine to a solution of [Cd(S(2)CNHR)(2)]. This confirms that C-S bond cleavage must be coupled to deprotonation of the -NH group, and explains why dialkylated derivatives [Cd(S(2)CNR(2))(2)] are inert to this particular mode of C-S bond cleavage. This system thus constitutes an unusual example of heterolytic, nonoxidative C-S bond cleavage that appears to proceed by a topochemical transfer of protons, which has implications for C-S bond cleavage processes in single-source precursors for II VI semiconductor materials. PMID- 15132626 TI - Methyl Tin(IV) derivatives of HOTeF(5) and HN(SO(2)CF(3))(2): a solution multinuclear NMR study and the X-ray crystal structures of (CH(3))(2)SnCl(OTeF(5)) and [(CH(3))(3)Sn(H(2)O)(2)][N(SO(2)CF(3))(2)]. AB - The new tin(IV) species (CH(3))(2)SnCl(OTeF(5)) was prepared via either the solvolysis of (CH(3))(3)SnCl in HOTeF(5) or the reaction of (CH(3))(3)SnCl with ClOTeF(5). It was characterized by NMR and vibrational spectroscopy, mass spectrometry, and single crystal X-ray diffraction. (CH(3))(2)SnCl(OTeF(5)) crystallizes in the monoclinic space group P2(1)/n (a = 5.8204(8) A, b =10.782(1) A, c =15.493(2) A, beta = 91.958(2) degrees, V = 971.7(2) A(3), Z = 4). NMR spectroscopy of (CH(3))(3)SnX, prepared from excess Sn(CH(3))(4) and HX (X = OTeF(5) or N(SO(2)CF(3))(2)), revealed a tetracoordinate tin environment using (CH(3))(3)SnX as a neat liquid or in dichloromethane-d(2) (CD(2)Cl(2)) solutions. In acetone-d(6) and acetonitrile-d(3) (CD(3)CN) solutions, the tin atom in (CH(3))(3)SnOTeF(5) was found to extend its coordination number to five by adding one solvent molecule. In the strong donor solvent DMSO, the Sn-OTeF(5) bond is broken and the (CH(3))(3)Sn(O=S(CH(3))(2))(2)(+) cation and the OTeF(5)(-) anion are formed. (CH(3))(3)SnOTeF(5) and (CH(3))(3)SnN(SO(2)CF(3))(2) react differently with water. While the Te-F bonds in the OTeF(5) group of (CH(3))(3)SnOTeF(5) undergo complete hydrolysis that results in the formation of [(CH(3))(3)Sn(H(2)O)(2)](2)SiF(6), (CH(3))(3)SnN(SO(2)CF(3))(2) forms the stable hydrate salt [(CH(3))(3)Sn(H(2)O)(2)][N(SO(2)CF(3))(2)]. This salt crystallizes in the monoclinic space group P2(1)/c (a = 7.3072(1) A, b =13.4649(2) A, c =16.821(2) A, beta = 98.705(1) degrees, V = 1636.00(3) A(3), Z = 4) and was also characterized by NMR and vibrational spectroscopy. PMID- 15132627 TI - Dendrimers with a copper(I) bis(phenanthroline) core: synthesis, electronic properties, and kinetics. AB - The copper(I) bis(chelate) complex Cu(L(0))(2) has been prepared from 2,9 diphenethyl-1,10-phenanthroline and Cu(CH(3)CN)(4)BF(4). Derivative Cu(L(0))(2) has been characterized by NMR, UV-vis spectroscopy, and X-ray crystallography. Interestingly, owing to the presence of the ethylene linker, the interligand pi pi stacking interactions between the phenyl rings and the phenanthroline subunits in Cu(L(0))(2) do not induce significant distortions of the pseudotetrahedral symmetry around the Cu(I) center in the solid state or in solution. Following the synthesis of Cu(L(0))(2), dendrimers Cu(L(1)(-)(4))(2) with a Cu(I) bis(2,9 diphenethyl-1,10-phenanthroline) core surrounded by Frechet type dendritic branches have been prepared and the kinetics of their cyanide-assisted demetalation studied. Importantly, the surrounding dendritic wedges have no significant influence on the coordination geometry of the Cu(I) center, as deduced from their absorption spectra. Therefore, the variations of the rate constants only reflect changes resulting from the presence of the dendritic branches. The kinetics of the cyanide-mediated demetalation reaction indeed revealed that cyanide diffusion through the dendritic shell is slightly influenced by the size of the branches. Significant effects were observed in the kinetics when going from the third to the fourth generation and have been ascribed to changes in the lipophilicity around the metallic core as a result of dendritic encapsulation. PMID- 15132628 TI - Crystallographic, electronic, and magnetic studies of zeta(2)-GaM (M = Cr, Mn or Fe): trends in itinerant magnetism. AB - This study of the crystal structure, electronic structure, and magnetic properties of the zeta(2)-GaM (M = Cr, Mn or Fe) alloys is motivated by the recent reinvestigation of the crystallographic Al(8)Cr(5) structure type of zeta(2)-GaMn. The isostructural compounds zeta(2)-GaFe and zeta(2)-GaCr have been refined using X-ray powder diffraction as well as neutron powder diffraction for zeta(2)-GaFe. Their structures have been refined using the space group Rm, with cell parameters a = 12.625(8) A and c = 7.785(10) A for zeta(2)-GaCr and a = 12.4368(11) A and c = 7.7642(10) A for zeta(2)-GaFe. Band structure calculations using the self-consistent, spin-polarized TB-LMTO method were performed to understand their electronic structure and magnetic properties. Band calculations show that from GaCr to GaFe the magnetic interactions change from weakly antiferromagnetic coupling to ferromagnetic coupling. Magnetic measurements confirm ferromagnetism for GaFe and show a weak paramagnetic response for GaCr. PMID- 15132629 TI - On the road to a termolecular complex with acetone: a heterometallic supramolecular network [[Rh(2)(O(2)CCF(3))(4)].micro(2) OCMe(2).[Cu(4)(O(2)CCF(3))(4)]]. AB - A novel heterometallic supramolecular network [[Rh(2)(O(2)CCF(3))(4)].micro(2) OCMe(2).[Cu(4)(O(2)CCF(3))(4)]](2)( infinity ) has been prepared by codeposition of the volatile mono(acetone) adduct [Rh(2)(O(2)CCF(3))(4).eta(1)-OCMe(2)](2) and copper(I) trifluoroacetate, [Cu(4)(O(2)CCF(3))(4)]. The product is of interest from the viewpoints of gas-phase supramolecular synthesis and a rare bridging coordination mode of acetone. It has been fully characterized by IR and NMR spectroscopy, elemental analysis, and X-ray diffraction. An X-ray structure revealed a layered 2D arrangement of the heterometallic [[Rh(2)(O(2)CCF(3))(4)].micro(2)-OCMe(2).[Cu(4)(O(2)CCF(3))(4)]] units built by axial intermolecular interactions of the open electrophilic Rh(II) and Cu(I) centers and O-atoms of neighboring carboxylate groups. The coordination of the acetone molecules within the [[Rh(2)(O(2)CCF(3))(4)].micro(2) OCMe(2).[Cu(4)(O(2)CCF(3))(4)]] unit is asymmetric with the Rh-O and Cu-O distances being 2.2173(15) and 2.7197(17) A, respectively. This work shows the potential of gas-phase deposition that may provide additional possibilities in supramolecular synthesis by utilizing intermolecular interactions and coordination bonds in a new way compared with conventional solution chemistry. PMID- 15132630 TI - An evaluation by density functional theory of M-M interactions in organometallic clusters with the [Fe(3)MoS(3)](2+) cores. AB - Density functional theory calculations were carried out on the structurally characterized [(Cl(4)-cat)Mo(py)Fe(3)S(3) (CO)(4)(P(n)Pr(3))(3)], A, and (Cl(4) cat)Mo(py)Fe(3)S(3)(CO)(6)(PEt(3))(2), B, and also on A(2)(-) and B(2+) clusters. The Fe-Fe distances in these molecules depend on the total number of valence electrons (60 e(-) in A and B(2)(+) and 62 e(-) in A(2)(-) and B) and undergo great structural changes upon addition or removal of electrons. The changes are consistent with known electron-counting rules in organometallic chemistry. The weak nature of the Fe-Fe bonding interactions in these clusters is apparent in the very similar energies of states with widely different Fe-Fe distances. PMID- 15132631 TI - Unusually high chemical compressibility of normally rigid type-I clathrate framework: synthesis and structural study of Sn(24)P(19.3)Br(x)I(8)(-)(x) solid solution, the prospective thermoelectric material. AB - A novel tin phosphide bromide, Sn(24)P(19.3(2))Br(8), and Sn(24)P(19.3(2))Br(x)()I(8)(-)(x) (x = 0-8) solid solution have been prepared and structurally characterized. All compounds crystallize with the type-I clathrate structure in the cubic space group Pmn (No. 223). The clathrate framework of the title solid solution shows a remarkable chemical compressibility: the unit cell parameter drops from 10.954(1) to 10.820(1) A on going from x = 0 to x = 8, a feature that has never been observed for normally rigid clathrate frameworks. The chemical compressibility as well as non-Vegard dependence of the unit cell parameter upon the bromine content is attributed to the nonuniform distribution of the guest halogen atoms in the polyhedral cavities of the clathrate framework. The temperature-dependent structural study performed on Sn(24)P(19.3(2))Br(8) has shown that, in contrast to the chemical compressibility, the thermal compressibility (linear contraction) of the phase is similar to that observed for the Group 14 anionic clathrates. The tin phosphide bromide does not undergo phase transition down to 90 K, and the atomic displacement parameters for all atoms decrease linearly upon lowering the temperature. These linear dependencies have been used to assess such physical constants as Debye temperature, 220 K, and the lattice part of thermal conductivity, 0.7 W/(m K). Principal differences between the title compounds and the group 14 anionic clathrates are highlighted, and the prospects of creating new thermoelectric materials based on cationic clathrates are briefly discussed. PMID- 15132632 TI - The bis-phenyltin-substituted, lone-pair-containing tungstoarsenate [(C(6)h(5)Sn)(2)as(2)W(19)O(67)(H(2)O)](8)(-). AB - The bis-phenyltin-substituted, lone-pair-containing tungstoarsenate [(C(6)H(5)Sn)(2)As(2)W(19)O(67)(H(2)O)](8)(-) (1) has been synthesized and characterized by multinuclear NMR, IR, and elemental analysis. Single-crystal X ray analysis was carried out on (NH(4))(7)Na[(C(6)H(5)Sn)(2)As(2)W(19)O(67)(H(2)O)].17.5H(2)O (NH(4)(-1), which crystallizes in the monoclinic system, space group P2(1)/c, with a = 18.3127(17) A, b = 24.403(2) A, c = 22.965(2) A, beta = 106.223(2) degrees, and Z = 4. Polyanion 1 consists of two B-alpha-(As(III)W(9)O(33)) Keggin moieties linked via a WO(H(2)O) fragment and two SnC(6)H(5) groups leading to a sandwich-type structure with nominal C(2)(v) symmetry. Polyanion 1 is stable in solution as indicated by the expected 6-line pattern (4:4:4:4:2:1) in (183)W NMR and the expected (119)Sn, (13)C, and (1)H NMR spectra. Synthesis of 1 was accomplished by reaction of C(6)H(5)SnCl(3) and K(14)[As(2)W(19)O(67)(H(2)O)] in a 2:1 molar ratio in aqueous acidic medium (pH 2). In the solid-state structure of NH(4)(-1, neighboring polyanions are weakly bound via W-O-Na bonds leading to chains which interact with each other via the phenyl rings resulting in a 2-D assembly. PMID- 15132633 TI - Synthesis of [NH(4)]MnCl(2)(OAc) and [NH(4)](2)MnCl(4)(H(2)O)(2) by solvothermal dehydration and structure/property correlations in a one-dimensional antiferromagnet. AB - The utility of the solvothermal dehydration strategy whereby superheated acetonitrile reacts with water of hydration to form ammonium acetate is demonstrated in the synthesis of [NH(4)]MnCl(2)(OAc), I, and [NH(4)](2)MnCl(4)(H(2)O)(2), II, from MnCl(2).4H(2)O. The structure of I is shown to crystallize in the monoclinic space group C2/c (No. 15) with a = 15.191(6) A, b = 7.044(2) A, c = 13.603(6) A, beta = 107.31 degrees, V = 1389.7(9) cm(-)(1), and Z = 8. The structure of II crystallizes in the space group I4/mmm (No. 139) with a = 7.5250(5) A, b = 8.276(2) A, V = 468.6(1) cm(-)(1), and Z = 2. Both structures exhibit extensive hydrogen bonding that controls both local Mn-Cl bonding and the interchain organization. I is shown to be a one-dimensional Heisenberg antiferromagnet with an intrachain exchange constant J/k = -2.39 K. This structure exhibits exchange coupling intermediate between the well-studied triply and doubly chloride-bridged one-dimensional manganese Heisenberg antiferromagnets. The structure/property correlation demonstrates a linear dependence of the exchange constant on the Mn-Cl-Mn bond angle, alpha, for alpha < 94 degrees. PMID- 15132634 TI - An electron localization function study of the geometry of d(0) molecules of the period 4 metals Ca to Mn. AB - We have studied the geometry of the formally d(0) MX(n)() (X = F, H, CH(3) and O; n = 2-6) molecules of the period 4 metals from Ca to Mn by studying the topology of the electron localization function (ELF) in order to try to understand why many of these molecules have non-VSEPR geometries. The quantitative analysis of the core basin population shows that it is always larger than its conventional value (18) because, in the LCAO-MO scheme, the 3d basis functions centered on the metal noticeably contribute to the electron density within the core region associated with the M shell. Therefore, the density available to form the bonds is less than Z(M) - 18, the value adopted in electron counts. Under the influence of the ligands, these electrons cause the core to lose its spherical symmetry by the formation of opposite-spin pair localization basins, which in turn influence the geometry of the ligands if the interaction of the ligands with the core is sufficiently strong. All of the ligands considered in this study, except F, interact with the core sufficiently strongly to give non-VSEPR geometries, which we have rationalized on the basis of the ELF topology. PMID- 15132635 TI - Multi-iron wells-Dawson heteropolytungstates. Electrochemical probing of siderophoric behavior in sandwich-type complexes. AB - The demetalation process of 10 multi-iron Wells-Dawson polyoxometalates is studied by cyclic voltammetry and controlled potential coulometry. Eight sandwich type complexes (alphaalphaalphaalpha Na(16)[(NaOH(2))(2)(Fe(III))(2)(X(2)W(15)O(56))(2)], alphaalphabetaalpha Na(14)[(NaOH(2))(Fe(III)OH(2))(Fe(III))(2)(X(2)W(15)O(56))(2)], alphabetabetaalpha-Na(12)[(Fe(III)OH(2))(2)(Fe(III))(2)(X(2)W(15)O(56))(2)], and alphabetabetaalpha-Na(14)[(Mn(II)OH(2))(2)(Fe(III))(2)(X(2)W(15)O(56))(2)] (where X = P(V) or As(V))) and two monomeric complexes (alpha Na(11)[(P(2)(Fe(III)Cl)(2)(Fe(III)OH(2))W(15)O(59))] and alpha Na(11)[(As(2)(Fe(III)Cl)(2)Fe(III)OH(2))W(15)O(59))]) were selected for this study. All 10 complexes show Fe(III) waves which are well-separated from the redox activity of the W(VI) centers. At room temperature and under mild conditions, iron release from the complexes is observed upon reduction of the Fe(III) centers. This release is controlled by the ionic strength of the medium, the nature and concentration of the anions present in the supporting electrolyte, and by the pH of the solution. This behavior parallels those described for most siderophores which depend on the same parameters. PMID- 15132636 TI - Ternary rare earth osmium aluminides R(7+)(x)Os(12)Al(61+)(y) belonging to a structural family with layered topology. AB - The 10 intermetallic compounds R(7+)(x)Os(12)Al(61+)(y) (R = Y, Nd, Sm, Gd-Tm) were prepared by arc-melting of the elemental components. They crystallize with a hexagonal structure very similar to that of Y(7.28)Re(12)Al(61.38). The structure was determined from four-circle diffractometer data of Y(7+)(x)Os(12)Al(61+)(y): P6(3)/mcm, a = 1301.5(2) pm, c = 903.0(2) pm, Z =1. Four atomic sites, all located on the 6(3) axis, show fractional occupancy, resulting in the composition Y(7.86(1))Os(12)Al(61.51(4)), corresponding to the Pearson symbol hP90-8.63. The structure may be viewed as consisting of alternating atomic layers of two kinds, although chemical bonding within and between the layers is of similar character as can be judged from the near-neighbor environments, where all of the 11 atomic sites have high coordination numbers. One kind of layers (A). is relatively loosely packed and contains the yttrium and some aluminum atoms. The other kind (B). consists of the osmium and the remaining aluminum atoms in a nearly hexagonal close-packed arrangement. These layers are stacked in the sequence ABAB. A similar building principle has recently been recognized for several other structures of ternary intermetallic compounds of rare earth and transition metals with a high content of aluminum or gallium, where the structures of CeOsGa(4), Ho(3)Ru(4)Ga(15), and Y(2)Pt(6)Al(15) are the most recent examples. This structural family is briefly reviewed. The cell volume of Yb(7+)(x)Os(12)Al(61+)(y) indicates a mixed or intermediate valence character +2/+3 for the ytterbium atoms of this compound. PMID- 15132637 TI - Cyano-bridged 2D Cu(II)-Cr(III) coordination polymers: structural evidence for formation of a polymeric macrocyclic metallic compound. AB - Two unique cyano-bridged 2D coordination polymers have been synthesized and characterized structurally and magnetically. The complexes contain two polyaza Cu(II) units and one novel macromolecular Cu(II) moiety, which have been synthesized via one-pot metal template condensation reactions involving ethylenediamine (en) and formaldehyde. Self-assembly of the polyaza Cu(II) mixture with [Cr(CN)(6)](3)(-) gave rise to two layered complexes. One complex contains unprecedented covalently linked polymeric Cu(II) chains and cyano bridged Cu(II)(-)Cr(III) coordination chains, which are interwoven to form a novel layer. The other complex shows intriguing encapsulation of [Cr(CN)(6)](3)( ) anions. Intermetallic ferromagnetic coupling is operative within the bridged 2D layer. The magnetic susceptibilities of both complexes were simulated using approximate models. PMID- 15132638 TI - A hybrid density functional study of O-O bond cleavage and phenyl ring hydroxylation for a biomimetic non-heme iron complex. AB - Density functional calculations using the B3LYP functional have been used to study the reaction mechanism of [Fe(Tp(Ph2))BF] (Tp(Ph2) = hydrotris(3,5 diphenylpyrazol-1-yl)borate; BF = benzoylformate) with dioxygen. This mononuclear non-heme iron(II) complex was recently synthesized, and it proved to be the first biomimetic complex reproducing the dioxygenase activity of alpha-ketoglutarate dependent enzymes. Moreover, the enthalpy and entropy of activation for this biologically interesting process were derived from kinetic experiments offering a unique possibility for direct comparison of theoretical and experimental data. The results reported here support a mechanism in which oxidative decarboxylation of the keto acid is the rate-limiting step. This oxygen activation process proceeds on the septet potential energy surface through a transition state for a concerted O-O and C-C bond cleavage. In the next step, a high-valent iron-oxo species performs electrophilic attack on the phenyl ring of the Tp(Ph2) ligand leading to an iron(III)-radical sigma-complex. Subsequent proton-coupled electron transfer yields an iron(II)-phenol intermediate, which can bind dioxygen and reduce it to a superoxide radical. Finally, the protonated superoxide radical leaves the first coordination sphere of the iron(III)-phenolate complex and dismutates to dioxygen and hydrogen peroxide. The calculated activation barrier (enthalpy and entropy) and the overall reaction energy profile agree well with experimental data. A comparison to the enzymatic process, which is suggested to occur on the quintet surface, has been made. PMID- 15132639 TI - Electrochemical investigations of the [tris(2 (diphenylphosphino)thiaphenolato)ruthenate(II)] monoanion reveal metal- and ligand-centered events: radical, reactivity, and rate. AB - Electrochemical investigations of [bis(triphenylphosphoranylidene)ammonium)][tris(2 (diphenylphosphino)thiaphenolato)ruthenate(II)], PPN[Ru(DPPBT)(3)] (1), and [(bis(2-(diphenylphosphino)thiaphenolato)methane)(2 (diphenylphosphino)thiaphenolato)ruthenium(II)] chloride, [Ru((DPPBT)(2)CH(2))(DPPBT)]Cl (2) are reported. Complex 1 is oxidized reversibly in a metal-centered event by one electron at a potential of +455 mV (vs Ag/AgCl) to the ruthenium(III) derivative [tris(2 (diphenylphosphino)thiaphenolato)ruthenium(III)], 3. Complex 3 can also be prepared by iodine oxidation of 1 in acetonitrile. Oxidation of 3 in acetonitrile is reversible on a cyclic voltammetry time scale but irreversible upon bulk oxidation yielding Ru-X. Monitoring the oxidation of 3 by UV-visible spectroscopy reveals a proposed metal-coordinated thiyl radical intermediate with a maximum absorbance at 850 nm. This intermediate decays at a temperature of -20 degrees C with a rate constant of (5.82 +/- 0.73) x 10(-)(3) s(-)(1) with a small, positive deltaH and a large, negative deltaS. Ru-X can be oxidized reversibly to Ru-Y at a potential of +806 mV but cannot be reduced. Complex 2 is reversibly oxidized by one electron in a metal-centered event to 4 at a potential of +767 mV. PMID- 15132640 TI - Toward a general strategy for the synthesis of heterobimetallic coordination complexes for use as precursors to metal oxide materials: synthesis, characterization, and thermal decomposition of Bi(2)(Hsal)(6).M(acac)(3) (M = Al, Co, V, Fe, Cr). AB - Bismuth(III) salicylate, [Bi(Hsal)(3)](n), reacts readily with the trivalent metal beta-diketonate compounds M(acac)(3) (acac = acetylacetonate; M = Al, V, Cr, Fe, Co) to produce trinuclear coordination complexes of the general formula Bi(2)(Hsal)(6).M(acac)(3) (M = Al, V, Cr, Fe, Co) in 60-90% yields. Spectroscopic and single crystal X-ray diffraction experiments indicate that these complexes possess an unusual asymmetric nested structure in both solution and solid state. Upon standing in dichloromethane solution, Bi(2)(Hsal)(6).Co(acac)(3) eliminates Bi(Hsal)(3) to give the 1:1 adduct Bi(Hsal)(3).Co(acac)(3). The 2:1 heterobimetallic molecular compounds undergo facile thermal decomposition on heating in air to 475 degrees C to produce heterometallic oxide materials, which upon annealing for 2 h at 700 degrees C form crystalline oxide materials. The synthetic approach detailed here represents a unique, general approach to the formation of heterobimetallic bismuth-based coordination complexes via the coordination of M(acac)(3) complexes to bismuth(III) salicylate. PMID- 15132641 TI - Low-coordinate iron(II) amido complexes of beta-diketiminates: synthesis, structure, and reactivity. AB - The synthesis, structure, and reactivity of a series of low-coordinate Fe(II) diketiminate amido complexes are presented. Complexes L(R)FeNHAr (R = methyl, tert-butyl; Ar = para-tolyl, 2,6-xylyl, and 2,6-diisopropylphenyl) bind Lewis bases to give trigonal pyramidal and trigonal bipyramidal adducts. In the adducts, crystallographic and (1)H NMR evidence supports the existence of agostic interactions in solid and solution states. Complexes L(R)FeNHAr may be oxidized using AgOTf, and the products L(R)Fe(NHAr)(OTf) are characterized with (19)F NMR spectroscopy, UV/vis spectrophotometry, solution magnetic measurements, elemental analysis, and, in one case, X-ray crystallography. In the structures of the iron(III) complexes L(R)Fe(NHAr)(OTf) and L(R)Fe(OtBu)(OTf), the angles at nitrogen and oxygen result from steric effects and not pi-bonding. The reactions of the amido group of L(R)FeNHAr with weak acids (HCCPh and HOtBu) are consistent with a basic nitrogen atom, because the amido group is protonated by terminal alkynes and alcohols to give free H(2)NAr and three-coordinate acetylide and alkoxide complexes. The trends in complex stability give insight into the relative strength of bonds from three-coordinate iron to anionic C-, N-, and O donor ligands. PMID- 15132643 TI - Analysis of major ampullate silk cDNAs from two non-orb-weaving spiders. AB - Compared to other arthropods, spiders are unique in their use of silk throughout their life span and the extraordinary mechanical properties of the silk threads they produce. Studies on orb-weaving spider silk proteins have shown that silk proteins are composed of highly repetitive regions, characterized by alanine and glycine-rich units. We have isolated and sequenced four partial cDNA clones representing major ampullate spider silk gene transcripts from two non-orb weavers: three for Kukulcania hibernalis and one for Agelenopsis aperta. These cDNA sequences were compared to each other, as well as to the previously published orb-weaver silk gene sequences. The results indicate that the repeats encoding conserved amino acid motifs such as polyA and polyGA that are characteristic of some orb-weaving spider silks are also found in some of the cDNAs reported in this study. However, we also found other motifs such as polyGS and polyGV in the cDNA sequences from the two non-orb-weaving spiders. The amino acid composition of the silk gland extracts shows that alanine and glycine are the major components of the silk of these two non-orb weavers as is the case in orb-weaver silks. Sequence alignment shows that A. aperta's cDNA displays a C terminal encoding region that is about 44% similar to the one present in N. clavipes's MaSp1 cDNA. In addition, as previously observed for spider silk sequences, the analysis of the codon usage for these four cDNAs demonstrates a bias for A or T in the wobble base position. PMID- 15132644 TI - A DECODER NMR study of backbone orientation in Nephila clavipes dragline silk under varying strain and draw rate. AB - Using DECODER (direction exchange with correlation for orientation distribution evaluation and reconstruction) NMR, we probe the orientations of carbonyl carbons in [1-(13)C]glycine-labeled dragline silk under conditions of varying strain and fiber draw rate. A model-specific reconstruction of the molecular orientation distribution incorporating beta sheets and polyglycine II helices indicates that the structures' alignment along the fiber can be described by a pair of Gaussian distributions with full width at half-maxima of 20 and 68 degrees and approximately 45 and approximately 55% relative contributions to the signal intensity. The alignment along the fiber was found to change appreciably when the drawing tension on the fiber was relaxed in a sample drawn at 4 cm/s while little change was observed in a sample drawn at 2 cm/s. The degree of alignment along the fiber was found to increase with fiber draw rate. PMID- 15132645 TI - Construction of silk fiber core in lepidoptera. AB - The formation and properties of lepidopteran silk fibers depend on amino acid repeats in the principal protein, heavy chain fibroin (H-fibroin). In H-fibroins of the "bombycoid" type, concatenations of alanine or of the GAGAGS crystalline motifs (1st tier repeats) and adjacent sequences breaking periodicity make 2nd tier repeats. Two to six such repeats comprise a 3rd tier assembly, and 12 assemblies, linked by an amorphous sequence, constitute the repetitive H-fibroin region. Heterogeneity in the repeat length and intercalation of amorphous regions prevent excessive crystallization. In the "pyraloid" H-fibroins, iterations of simple motifs are absent and assemblies of several complex motifs constitute highly regular repeats that are organized in about 12 highest order reiterations without specific spacers. Repeat homogeneity appears crucial for the alignment and interaction of the disjunct motifs that must be registered precisely to form crystallites; repeat heterogeneity is associated with decreased fiber strength. Both H-fibroin types are typically hydrophobic, and their secretion requires disulfide linkage to light chain fibroin and participation of another protein, P25. These auxiliary proteins are absent in saturniid moths with amphiphilic H fibroin repeats. The selection at nucleic acid and protein levels and the availability of nutrients play roles in H-fibroin evolution. PMID- 15132646 TI - Supercontraction stress in spider webs. AB - Silk produced from the major ampullate (MA) gland supercontracts when wet, and in this paper, we investigate the consequences of high humidity and of the added load of water droplets condensing from saturated air on the mechanical integrity of the spiders' orb web. We measured the development of the supercontraction stress (sigma(sc)) with time when fixed lengths of MA silk from Nephila clavipes and Argiope aurantia were exposed to increasing humidity. Supercontraction generated stresses of about 50 MPa, and extension of these samples to stresses between 150 and 1100 MPa show a time dependent relaxation over 1000 s to approximately 75% of the initial tension but show no indication of failure. We conclude that supercontraction can maintain tension in webs and does not limit the ability of the web to support loads in excess of the supercontraction stress. PMID- 15132647 TI - Structures of Bombyx mori and Samia cynthia ricini silk fibroins studied with solid-state NMR. AB - There are many kinds of silks spun by silkworms and spiders, which are suitable to study the structure-property relationship for molecular design of fibers with high strength and high elasticity. In this review, we mainly focus on the structural determination of two well-known silk fibroin proteins that are from the domesticated silkworm, Bombyx mori, and the wild silkworm, Samia cynthia ricini, respectively. The structures of B. mori silk fibroin before and after spinning were determined by using an appropriate model peptide, (AG)(15), with several solid-state NMR methods; (13)C two-dimensional spin-diffusion solid-state NMR and rotational echo double resonance (REDOR) NMR techniques along with the quantitative use of the conformation-dependent (13)C CP/MAS chemical shifts. The structure of S. c. ricini silk fibroin before spinning was also determined by using a model peptide, GGAGGGYGGDGG(A)(12)GGAGDGYGAG, which is a typical repeated sequence of the silk fibroin, with the solid-state NMR methods. The transition from the structure of B. mori silk fibroin before spinning to the structure after spinning was studied with molecular dynamics calculation by taking into account several external forces applied to the silk fibroin in the silkworm. PMID- 15132648 TI - Molecular and mechanical properties of major ampullate silk of the black widow spider, Latrodectus hesperus. AB - Molecular and material properties of major ampullate silk were studied for the cobweb-building black widow spider Latrodectus hesperus. Material properties were measured by stretching the silk to breaking. The strength was 1.0 +/- 0.2 GPa, and the extensibility was 34 +/- 8%. The secondary structure of the major ampullate silk protein was studied using carbon-13 NMR spectroscopy. Alanine undergoes a transition from a coiled structure in pre-spun silk to a beta sheet structure in post-spun silk. We have also isolated two distinct cDNAs (both about 500 bp) which encode proteins similar to major ampullate spidroin 1 and 2 (MaSp1 and MaSp2). The MaSp1-like silk contains polyalanine runs of 5-10 residues as well as GA and GGX motifs. The MaSp2-like silk contains polyalanine runs of varying length as well as GPG(X)(n) motifs. L. hesperus major ampullate silk is more like major ampullate silk from other species than other L. hesperus silks. PMID- 15132649 TI - X-ray evidence for a "super"-secondary structure in silk fibers. AB - X-ray studies on degummed B. mori silk fibers and on hydrogels prepared under a variety of conditions reveal moderately small angle reflections. These reflections are often highly oriented and are correlated to silk II lattice reflections. A superstructure can explain these features. Silk fibroin hydrogels were monitored as they dried to form the silk II structure. The silk II wide angle and moderately small angle patterns obtained from dried hydrogels and silk fibers are identical. The "superstructure" reflections at moderately small angle (3-7 nm) were first to appear, followed by the "intersheet" spacing, and then the remainder of the silk II wide angle scattering pattern. Thus, any superstructure hypothesized for the hydrogels (and for Silk II in fibers) must be both stable in a highly hydrated environment and must convert to silk II with little large scale diffusion. A folded structure, similar to amyloids and cross-beta-sheets but with much longer beta-strand stems, is proposed for silk II in fibers. PMID- 15132650 TI - Spider silk protein refolding is controlled by changing pH. AB - Spidroins, the major silk proteins making up the spider's dragline silk, originate in two distinct tissue layers (A and B) in the spider's major ampullate gland. Formation of the complex thread from spidroins occurs in the lumen of the duct connected to the gland. Using pH-sensitive microelectrode probes, we showed that the spidroins traveling through the gland and duct experience a monotonic decrease in pH from 7.2 to 6.3. In addition, circular dichroism spectroscopy of material extracted from the gland showed a structural refolding concomitant with position in the gland and post-extraction changes in pH. We demonstrate that lowering the pH in vitro causes a dramatic conformational change in the protein from the A zone, converting it irreversibly from a coil to a predominantly beta sheet structure. Furthermore, amino acid analyses have indicated that there are at least two distinct, though similar, proteins secreted in the A and B zones suggesting a potential factor in the progressive acidification as well as a pH sensitivity of the folding of spidroins in the gland. Thus, we provide, for the first time, a quantitative map of the pH value and position correlated with molecular structural folding in the silk gland characterizing the crucial role that pH plays in spider silk formation. PMID- 15132651 TI - Biomaterial films of Bombyx mori silk fibroin with poly(ethylene oxide). AB - Phase separation into controllable patterned microstructures was observed for Bombyx mori silkworm silk and poly(ethylene oxide) (PEO) (900000 g/mol) blends cast from solution. The evolution of the microstructures with increasing PEO volume fraction is strikingly similar to the progression of phases and microstructures observed with surfactants. The chemically patterned materials obtained provide engineerable biomaterial surfaces with predictable microscale features which can be used to create topographically patterned or chemically functionalized biomaterials. Solution blending was used to incorporate water soluble PEO into silk to enhance elasticity and hydrophilicity. The sizes of the globule fibroin phase ranged from 2.1 +/- 0.5 to 18.2 +/- 2.1 microm depending on the ratio of silk/PEO. Optical microscopy and SEM analysis confirmed the micro phase separation between PEO and silk. Surface properties were determined by XPS and contact angle. Methanol can be used to control the conformational transition of silk fibroin to the insoluble beta-sheet state. Subsequentially, the PEO can be easily extracted from the films with water to generate silk matrixes with definable porosity and enhanced surface roughness. These blend films formed from two biocompatible polymers provide potential new biomaterials for tissue engineering scaffolds. PMID- 15132652 TI - Porous 3-D scaffolds from regenerated silk fibroin. AB - Three fabrication techniques, freeze-drying, salt leaching and gas foaming, were used to form porous three-dimensional silk biomaterial matrixes. Matrixes were characterized for morphological and functional properties related to processing method and conditions. The porosity of the salt leached scaffolds varied between 84 and 98% with a compressive strength up to 175 +/- 3 KPa, and the gas foamed scaffolds had porosities of 87-97% and compressive strength up to 280 +/- 4 KPa. The freeze-dried scaffolds were prepared at different freezing temperatures (-80 and -20 degrees C) and subsequently treated with different concentrations (15 and 25%) and hydrophilicity alcohols. The porosity of these scaffolds was up to 99%, and the maximum compressive strength was 30 +/- 2 KPa. Changes in silk fibroin structure during processing to form the 3D matrixes were determined by FT-IR and XrD. The salt leached and gas foaming techniques produced scaffolds with a useful combination of high compressive strength, interconnected pores, and pore sizes greater than 100 microns in diameter. The results suggest that silk-based 3D matrixes can be formed for utility in biomaterial applications. PMID- 15132653 TI - Consequences of forced silking. AB - The forced silking of a spider to obtain major ampullate (MA) silk for experiments is a standard practice; however, this method may have profound effects on the resulting silk's properties. Experiments were performed to determine the magnitude of the difference in the forces required to draw silk from the MA gland between unrestrained spiders descending on their draglines and restrained spiders from which MA silk was drawn with a motor. The results show that freely falling spiders can spool silk with as little as 0.1 body weights of force, which generates a stress that is about 2% of the silk's tensile strength. In contrast, forcibly silked spiders apply as much as 4 body weights of force with an internal braking mechanism, and this force creates silk stresses in excess of 50% of the silk's tensile strength. The large forces observed in forced silking should strongly affect the draw alignment of the polymer network in the newly spun fibers, and this may account for the differences in material properties observed between naturally spun and forcibly spun MA silks. In addition, the heat produced by the internal friction brake during forced silking may set the upper limit of forced silking speed. PMID- 15132654 TI - Variation of mechanical properties with amino acid content in the silk of Nephila clavipes. AB - In this paper, we explore the impact of dietary deprivation, where spiders are provided diets missing one or more of the amino acids, on the properties of the spider dragline silk spun after one month on the diet. Cohorts of female N. clavipes spiders were selected for diets deprived of alanine (Ala) and glycine (Gly), arginine (Arg), leucine (Leu), or tyrosine (Tyr), and their silk was harvested twice weekly during the one-month course of the diet. Significant mechanical differences are observed after as little as 6 days on the diet. Utilizing conventional tensile testing methods, single fibers were strained to break so as to study the influence of diet on the stress/strain properties. Diets deprived of Ala and Gly appear to most directly impact the load-bearing foundation of dragline silk. Diets deprived of Arg, Tyr, and possibly Leu reduce the strength of the silk, and diets missing Tyr and Leu reduce the strain-to failure. Observations obtained from ESEM photos of the fracture interfaces after tensile testing illustrate the fracture mechanics of spider silk. Both solid state NMR and amino acid analysis of the digested protein suggest, however, that the relationship between diet and amino acid incorporation into the silk fiber is not straightforward. PMID- 15132655 TI - Broad patterns of speciation are correlated with the evolution of new silk proteins in spiders but not in the lepidoptera. AB - Silk synthesis is an ancestral character of both the Lepidoptera and the Araneae. Araneae evolution is marked by an increased commitment to a silk-producing physiology. At least three major Araneae speciation events are correlated with the evolution of new silk-producing glands and new silk proteins. In contrast, although 98% of the Lepidoptera produce silk, there appears to be no relationship between silk use, protein types, and species numbers. The differences in these two systems, both meeting a need to produce a large volume of protein, may reflect predictable resource availability to herbivorous Lepidoptera larvae but fluctuating resource availability to carnivorous spiders. PMID- 15132656 TI - Structure of the model peptides of Bombyx mori silk-elastin like protein studied with solid state NMR. AB - The peptides (AG)(6)(VPGVG)(AG)(7) and (AG)(5)(VPGVG)(2)(AG)(5) are models for a new type of protein with both composition and properties such as Bombyx mori silk and elastin. In this paper, we report the solid-state NMR results for these samples and related peptides; the structures after dialysis of the 9 M LiBr aqueous solution and after treatment with formic acid were determined and compared. The detailed structural analyses were performed using deconvolution subroutines assuming Gaussian line shapes for the Ala Cbeta peaks of the (AG)(n) sequences in these peptides. The peptide (AG)(6)(VPGVG)(AG)(7) took the silk II structure after the dialysis, which is in contrast to the silk I form of (AG)(15) after the same treatment. However, a drastic structural change of the (AG)(n) sequences was observed for (AG)(5)(VPGVG)(2)(AG)(5); the fraction of distorted beta-turn was 81% after the dialysis, but the distorted beta-sheet became dominant (84%) after treatment with formic acid. The local structures of the Gly residue of the VG units in the elastin-like subunits, (VPGVG) and (VPGVG)(2), were the distorted structures with a distribution of the torsion angles, which was derived from the 2D spin diffusion NMR spectral pattern of (AG)(5)VPG[1 (13)C]V[1-(13)C]GVPGVG(AG)(5). Observation of this distribution of the Gly residue was independent of the treatment, dialysis or formic acid. PMID- 15132657 TI - Phase behavior and hydration of silk fibroin. AB - The osmotic stress method was applied to study the thermodynamics of supramolecular self-assembly phenomena in crystallizable segments of Bombyx mori silkworm silk fibroin. By controlling compositions and phases of silk fibroin solution, the method provided a means for the direct investigation of microscopic and thermodynamic details of these intermolecular interactions in aqueous media. It is apparent that as osmotic pressure increases, silk fibroin molecules are crowded together to form silk I structure and then with further increase in osmotic pressure become an antiparallel beta-sheet structure, silk II. A partial ternary phase diagram of water-silk fibroin-LiBr was constructed based on the results. The results provide quantitative evidence that the silk I structure must contain water of hydration. The enhanced control over structure and phase behavior using osmotic stress, as embodied in the phase diagram, could potentially be utilized to design a new route for water-based wet spinning of regenerated silk fibroin. PMID- 15132658 TI - Structural conformation of spidroin in solution: a synchrotron radiation circular dichroism study. AB - Spider silk is made and spun in a complex process that tightly controls the conversion from soluble protein to insoluble fiber. The mechanical properties of the silk fiber are modulated to suit the needs of the spider by various factors in the animal's spinning process. In the major ampullate (MA) gland, the silk proteins are secreted and stored in the lumen of the ampulla. A particular structural fold and functional activity is determined by the spidroins' amino acid sequences as well as the gland's environment. The transition from this liquid stage to the solid fiber is thought to involve the conversion of a predominantly unordered structure to a structure rich in beta-sheet as well as the extraction of water. Circular dichroism provides a quick and versatile method for examining the secondary structure of silk solutions and studying the effects of various conditions. Here we present the relatively novel technique of synchrotron radiation based circular dichroism as a tool for investigating biomolecular structures. Specifically we analyze, in a series of example studies on structural transitions induced in liquid silk, the type of information accessible from this technique and any artifacts that might arise in studying self-assembling systems. PMID- 15132659 TI - pH induced changes in the rheology of silk fibroin solution from the middle division of Bombyx mori silkworm. AB - The rheological properties of fibroin silk solutions extracted from the middle division of Bombyx mori silkworms were examined. Acidification of the solutions with acetic acid vapor gelled the material, a process which at short time scales could be reversed by exposure to ammonia vapor. The solution could also be converted to sol from the gel state by the addition of EDTA. The possible mechanisms for gel formation in fibroin solutions is discussed as are the implications for the process of spinning silk fibers. PMID- 15132660 TI - Optical spectroscopy to investigate the structure of regenerated Bombyx mori silk fibroin in solution. AB - Fluorescence and circular dichroism spectroscopy were used to monitor the conformational transition of regenerated Bombyx mori silk fibroin (RSF) in aqueous solutions under different conditions. According to the analysis of fluorescence spectra using anilinonaphthalene-8-sulfonic acid magnesium salt (ANS) as an external probe, the destruction of the hydrophobic core prior to the secondary structure change suggests that this collapse may initiate the conformational transition from random coil to beta-sheet for RSF. The temperature dependence of the structural changes of RSF, detected by both fluorescence spectroscopy and circular dichroism, shows a reversible process upon heating and recooling, with the midpoint around 45 degrees C. The results also indicate that most of the tryptophan (Trp) residues contained in silk fibroin are concentrated on the surface of the unfolded protein. However, they will change their location in the highly ordered structure (e.g., becoming more homogeneous) with the conformational transition of silk fibroin. Moreover, our studies also suggest that the presence of water plays a crucial role during the structure changes of fibroin. PMID- 15132661 TI - Modeling of mechanical properties and structural design of spider web. AB - With a unique combination of strength and toughness among materials, spider silk is the model for engineering materials. This paper presents the stress-strain behavior of Nephila clavipes spider silk under tension, transverse compression, and torsional deformation obtained by a battery of micro testing equipment. The experimental results showed significantly higher toughness than the state-of-the art fibers in tension and in transverse compression. Higher shear modulus was also observed for the spider silk comparing to other liquid crystalline fibers such as aramid fibers. On the basis of the experimental results finite element analysis is used to simulate static and dynamic properties of spider web and to explore the role of both material properties and architectural design in its structural integrity and mechanical performance. PMID- 15132662 TI - Structure and properties of silk hydrogels. AB - Control of silk fibroin concentration in aqueous solutions via osmotic stress was studied to assess relationships to gel formation and structural, morphological, and functional (mechanical) changes associated with this process. Environmental factors potentially important in the in vivo processing of aqueous silk fibroin were also studied to determine their contributions to this process. Gelation of silk fibroin aqueous solutions was affected by temperature, Ca(2+), pH, and poly(ethylene oxide) (PEO). Gelation time decreased with increase in protein concentration, decrease in pH, increase in temperature, addition of Ca(2+), and addition of PEO. No change of gelation time was observed with the addition of K(+). Upon gelation, a random coil structure of the silk fibroin was transformed into a beta-sheet structure. Hydrogels with fibroin concentrations >4 wt % exhibited network and spongelike structures on the basis of scanning electron microscopy. Pore sizes of the freeze-dried hydrogels were smaller as the silk fibroin concentration or gelation temperature was increased. Freeze-dried hydrogels formed in the presence of Ca(2+) exhibited larger pores as the concentration of this ion was increased. Mechanical compressive strength and modulus of the hydrogels increased with increase in protein concentration and gelation temperature. The results of these studies provide insight into the sol gel transitions that silk fibroin undergoes in glands during aqueous processing while also providing important insight in the in vitro processing of these proteins into useful new materials. PMID- 15132663 TI - Thermal analysis of water in silk-elastinlike hydrogels by differential scanning calorimetry. PMID- 15132664 TI - Anticoagulant and antiplatelet agents: their clinical and device application(s) together with usages to engineer surfaces. AB - An essential aspect of the treatment of patients with cardiovascular disease is the use of anticoagulant and antiplatelet agents for the prevention of further ischaemic events and vascular death resulting from thrombosis. Aspirin and heparin have been the standard therapy for the management of such conditions to date. Recently, numerous more potent platelet inhibitors together with anticoagulant agents have been developed and tested in randomized clinical trials. This article reviews the current state of the art of antiplatelet and anticoagulant therapy in light of its potential clinical efficacy. It then focuses on the usages of these agents in order to improve the performance of clinical devices such as balloon catheters, coronary stents, and femoropopliteal bypass grafting and extra corporeal circuits for cardiopulmonary bypass. The article then goes on to look at the usage of these agents more specifically heparin, heparan, hirudin, and coumarin in the development of more biocompatible scaffolds for tissue engineering. PMID- 15132665 TI - Cross-linked aggregates of multimeric enzymes: a simple and efficient methodology to stabilize their quaternary structure. AB - In this manuscript, we show that the immobilization of proteins following the technique of cross-linked protein aggregates (CLEAS) may permit the stabilization of the most complex multimeric enzymes by preventing their dissociation. To illustrate that, we have first prepared CLEAS with two tetrameric catalases. Activity recovery was over 40%, and no protein subunit could be desorbed from the CLEAS after boiling in SDS. More interestingly, the enzyme stability, which in its soluble form strongly depends on the enzyme concentration, becomes fully independent of this parameter. This permitted the enzyme stability to greatly increase under diluted conditions. In fact, diluted CLEAs presented a higher stability than those of their glyoxyl derivatives counterparts, which were unable to fully stabilize the multimeric structure of these tetrameric enzymes PMID- 15132666 TI - Poly(N-(2-hydroxypropyl) methacrylamide mono/di lactate): a new class of biodegradable polymers with tuneable thermosensitivity. AB - A novel class of thermosensitive and biodegradable polymers, poly(N-(2 hydroxypropyl) methacrylamide mono/di lactate) (poly(HPMAm-mono/di lactate)), was synthesized. The cloud points (CP) of poly(HPMAm-monolactate) and poly(HPMAm dilactate) in water were 65 and 13 degrees C, respectively. The lower CP for poly(HPMAm-dilactate) is likely due the greater hydrophobicity of the dilactate side group over the monolactate side group. The CP of poly(HPMAm-monolactate-co HPMAm-dilactate) increased linearly with mol % of HPMA-monolactate, which demonstrates that the CP is tuneable by the copolymer composition. PMID- 15132667 TI - Large area two-dimensional B cell arrays for sensing and cell-sorting applications. AB - Regular arrays of nonadherent B cells over large areas were produced with the use of micropatterned molecular templates consisting of a newly designed poly(allylamine)-g-poly(ethylene glycol) polycation graft copolymer. Polymer-on polymer stamping (POPS) techniques were applied successfully to create micron scale patterns of the graft copolymer on negatively charged multilayer surfaces without losing resistance to the nonspecific adsorption of proteins. To generate templates for B cell arrays, the characteristics of the patterned surface were modified via introduction of surface biotinylation and specific protein adsorption. The qualities of B cell arrays resulting from each template suggest the binding strength between nonadherent B cells and the template surface is the controlling factor in the fabrication of clean and regular arrays of immobilized lymphocytes over large areas, which is critical in many bio-technological and immunological applications. PMID- 15132668 TI - Feasibility of polysaccharide hybrid materials for scaffolds in cartilage tissue engineering: evaluation of chondrocyte adhesion to polyion complex fibers prepared from alginate and chitosan. AB - The ideal cell-carrier material for cartilage regeneration should be one that closely mimics the natural environment in a living articular cartilage matrix. In the current study, we considered that alginate-based chitosan hybrid biomaterials could provide excellent supports for chondrocyte adhesion. To test this hypothesis, we investigated the adhesion behavior of rabbit chondrocytes onto an alginate polymer versus the adhesion of the chondrocytes onto some alginate-based chitosan hybrid polymer fibers in vitro. We demonstrated that the alginate-based chitosan hybrid polymer fibers showed much improved adhesion capacity with chondrocytes in comparison with alginate polymer fiber. Additionally, morphologic studies revealed maintenance of the characteristic round morphology of the chondrocyte and the dense fiber of the type II collagen produced by the chondrocytes in the hybrid polymer. On the basis of these results, we conclude that an alginate-based chitosan hybrid polymer fiber has considerable potential as a desirable biomaterial for cartilage tissue scaffolds. PMID- 15132669 TI - NMR characterization of native liquid spider dragline silk from Nephila edulis. AB - Solid spider dragline silk is well-known for its mechanical properties. Nonetheless a detailed picture of the spinning process is lacking. Here we report NMR studies on the liquid silk within the wide sac of the major ampullate (m.a.) gland from the spider Nephila edulis. The resolution in the NMR spectra is shown to be significantly improved by the application of magic-angle spinning (MAS). From the narrow width of the resonance lines and the chemical shifts observed, it is concluded that the silk protein within the wide sac of the m.a. gland is dynamically disordered throughout the molecule in the sense that each amino acid of a given type senses an identical environment, on average. The NMR data obtained are consistent with an isotropic liquid phase. PMID- 15132670 TI - Conserved C-termini of Spidroins are secreted by the major ampullate glands and retained in the silk thread. AB - The C-termini of Spidroins produced in the major and minor ampullate glands of spiders are highly conserved. Despite this conservation, no corresponding peptides have been identified in the spinning dopes or the silk filaments so far. To prove their presence or absence, polyclonal antibodies derived against fusion proteins containing the conserved C-terminal regions of both Spidroin 1 and 2 from the spider Nephila clavipes were generated. The antibodies reacted with high molecular weight polypeptides of the corresponding gland extracts and solubilized major ampullate filament and in addition to filament cross-sections. This demonstrates the existence of C-terminal specific peptides in the spinning dope and the mature Spidroins. Both the fusion proteins as well as the proteins contained within the gland lumen showed a reduction in their size under reducing conditions indicating the presence of disulfide bonds. Their high conservation and the biochemical data suggest crucial roles the C-termini play in the formation and/or structure of the corresponding silk filaments. PMID- 15132671 TI - Quantification of the effects of chain length and concentration on the thermal behavior of elastin-like polypeptides. AB - At a specific temperature, elastin-like polypeptides (ELPs) undergo a sharp solubility transition that can be exploited in a variety of applications in biotechnology and medicine. The temperature of the transition varies with ELP sequence, molecular weight, and concentration. We present a single equation of three parameters that quantitatively predicts the transition temperature as a function of ELP length and concentration for an ELP of a fixed composition. This model should be useful both for the design of new ELP sequences that exhibit a desired transition temperature and for the selection of variables to trigger the phase transition of an ELP for a given application. PMID- 15132672 TI - Co-aggregation of penicillin g acylase and polyionic polymers: an easy methodology to prepare enzyme biocatalysts stable in organic media. AB - A novel type of biocatalyst that combines the good properties of cross-linked enzyme aggregates (CLEAs) and hydrophilic microenvironments has been developed. Dextran sulfate- and polyethyleneimine-coated CLEAs of penicillin acylase (CLEA GDP) were prepared by adding the polymers of different sizes before the precipitation stage of the enzyme. This study presents the development and optimization of a protocol to produce such a biocatalyst using penicillin acylase as a model. Experiments show that CLEA-GDPs have a highly increased stability in organic media. The average half-life of the preparations was much higher than standard CLEA without a microenvironment (CLEA-G), (e.g., more than 25-fold) in the presence of dioxane. However, their thermal stability was not increased, which leads to the conclusion that the stability of CLEA-GDPs in organic media is due to the hydrophilic microenvironment that surrounds the protein enzyme more than to a conformational stiffening effect. This is further supported by solvation experiments that show a preferential hydration of CLEA when polymers are used to coat the enzyme. CLEA-GDPs are clearly better than other biocatalysts in terms of solvent stability. PMID- 15132673 TI - Regulation of enzyme-substrate complexation by a substrate conjugated with a phospholipid polymer. AB - To recognize and control ligand-receptor interactions at the interface between cells and polymer materials, we investigated a model system with an enzyme and a substrate conjugated with a biocompatible phospholipid polymer in an aqueous medium. We explored the regulation of enzyme-substrate (ES) complexation using horseradish peroxidase (HRP) as the enzyme and 4-aminoantipyrine (AAP) and 3-(p hydroxyphenyl) propionic acid (HPPA) as substrates. The phospholipid polymer (PMBN), composed of 2-methacryloyloxyethyl phosphorylcholine, n-butyl methacrylate, and p-nitrophenyloxycarbonyl poly(oxyethylene)methacrylate, was prepared and conjugated with AAP (PMBN-AAP conjugate). The formation and dissociation of the ES complex were investigated using capillary electrophoresis and fluorescence spectroscopy. In the chart of the capillary electrophoresis, a much longer retention time of HRP was observed in the PMBN-AAP conjugate-coated capillary compared with that in a nontreated capillary. The retention time was significantly longer in comparison with the case of a mixed solution of HRP and AAP. This result clearly shows that HRP forms an ES complex with the immobilized PMBN-AAP conjugate and that the addition of AAP to the medium inhibits the interactions between HRP and the PMBN-AAP conjugate. Though HRP forms an ES complex with both AAP and the PMBN-AAP conjugate, the ES complex with the PMBN AAP conjugate was easily dissociated by addition of HPPA as an alternative substrate because HRP started to react with the HPPA immediately. However, the HRP that formed an ES complex with AAP fell behind in reacting with the HPPA. The activity of HRP was maintained at the initial level in the presence of the PMBN AAP conjugate at 25 degrees C for 1 week. Additionally, even under H(2)O(2) conditions, HRP stored with the PMBN-AAP conjugate maintained 40% of the initial activity whereas HRP was deactivated within 6 h. This result indicates that the PMBN-AAP conjugate could block the active sites by formation of an ES complex. This is due to the formation of the ES complex, which retained the structure of HRP by blocking the active sites. On the basis of these results, we considered that the reversible attachment and detachment by PMBN conjugated with specific ligands from cellular receptors will be realized. PMID- 15132674 TI - Binding effect of Cu(2+) as a trigger on the sol-to-gel and the coil-to-helix transition processes of polysaccharide, gellan gum. AB - The binding effect of divalent cation Cu(2+) on the gelation process with a coil helix transition in Cu(2+)/gellan aqueous solutions has been successfully elucidated by EPR, CD, and viscoelasticity measurements. Generally, Na-type gellan gum in aqueous solution can make gel when accompanied by an intrinsic coil helix formation induced by hydrogen bonding between chains without any additional cations at T(ch)(-)(in) ( approximately 29 degrees C) with cooling temperature. An extrinsic coil-helix transition, induced by additional divalent cations in advance of the intrinsic sol-gel transition of gellan gum, is separately detected by CD measurement. The extrinsic coil-helix transition temperatures T(ch)(-)(ex) (>47 degrees C), which increased with the Cu(2+) concentration added, were nearly identical to the sol-gel transition temperature, T(sg), determined by the viscoelasticity measurement. Judging from the molar ellipticity by CD measurement and quantitative analysis of EPR spectra, it was elucidated that the helix forming process via divalent cations is composed of two steps ascribed to the different origins, i.e., a chemical binding effect via Cu(2+) ions in the initial stage and hydrogen bonds subsequently. Finally, we propose the coil-helix and the sol-gel transition mechanism initiated by the binding effect with the divalent cation, in which the partial chelate formation can cause local formation of helices and junction zones in the vicinity of the chelates at the initial stage of the process and stabilize the helices and the junction zones. On the other hand, the stabilized helices and junction zones can induce further formation and further stabilization of the Cu(2+)-gellan chelates. The mutual stabilization promotes the formation of three-dimensional network structure at the higher temperature than the intrinsic temperature for network formation. PMID- 15132675 TI - Thermodynamic studies on the adsorption of fibronectin adhesion-promoting peptide on nanothin films of poly(2-vinylpyridine) by SPR. AB - This paper describes a methodology for preparing uniform, nanothin polymer films for the study of biomolecule adsorption by surface plasmon resonance spectroscopy (SPR). The methodology combines molecular self-assembly of alkanethiols on gold with surface-confined atom transfer radical polymerization (ATRP). Poly(2 vinylpyridine) was chosen to demonstrate the methodology, and growth kinetics were studied by ex situ ellipsometry. Atomic force microscopy (AFM) indicated that the polymer films were uniform with RMS roughness of approximately 0.5 nm. Subsequent SPR measurements were done to determine thermodynamic adsorption properties (deltaG, deltaH, and deltaS) between fibronectin adhesion-promoting peptide and the surface-confined poly(2-vinylpyridine) at 15, 20, and 25 degrees C. The flexibility in synthesis conditions and the opportunities for manipulating film thicknesses and graft densities that ATRP provides to grow polymer films from gold surfaces holds advantages over conventional spin-coating and grafting to approaches in the design of model polymer films for biomolecule adsorption studies. These advantages are described. PMID- 15132676 TI - Permanent, nonleaching antibacterial surfaces. 1. Synthesis by atom transfer radical polymerization. AB - We have grown an antimicrobial polymer directly on the surfaces of glass and paper using atom transfer radical polymerization (ATRP). The method described here results in potentially permanent nonleaching antibacterial surfaces without the need to chemically graft the antimicrobial material to the substratum. The tertiary amine 2-(dimethylamino)ethyl methacrylate was polymerized directly onto Whatman #1 filter paper or glass slides via atom transfer radical polymerization. Following the polymerization, the tertiary amino groups were quaternized using an alkyl halide to produce a large concentration of quaternary ammonium groups on the polymer-modified surfaces. Incubating the modified materials with either Escherichia coli or Bacillus subtilis demonstrated that the modified surfaces had substantial antimicrobial capacity. The permanence of the antimicrobial activity was demonstrated through repeated use of a modified glass without significant loss of activity. Quaternary amines are believed to cause cell death by disrupting cell membranes allowing release of the intracellular contents. Atomic force microscopic imaging of cells on modified glass surfaces supports this hypothesis. PMID- 15132677 TI - Directed covalent immobilization of aminated DNA probes on aminated plates. AB - A new protocol that enables the immobilization of DNA probes on aminated micro titer plates activated with aldehyde-dextran via an amino group artificially introduced in the 3' end of the oligonucleotide probe is reported in this work. The method is based on the use of hetero-functional-dextran as a long and multifunctional spacer arm covalently attached to an aminated surface capable of immobilizing DNA oligonucleotides. The immobilization occurred only via the amino introduced in the 3' end of the probe, with no implication of the DNA bases in the immobilization, ensuring that the full length of the probe is available for hybridization. These plates having immobilized oligonucleotide probes are able to hybridize complementary DNA target molecules. The tailor-made hetero-functional aldehyde-aspartic-dextran together with the chemical blocking of the remaining primary amino groups on the support using acetic anhydride avoid the nonspecific adsorption of DNA on the surface of the plates. Using these activated plates, (studying the effect of the probe concentration, temperature, and time of the plate activation on the achieved signal), thus, the covalent immobilization of the aminated DNA probe was optimized, and the sensitivity obtained was similar to that achieved using commercial biotin-streptavidin systems. The new DNA plates are stable under very drastic experimental conditions (90% formamide, at 100 degrees C for 30 min or in 100 mM NaOH). PMID- 15132678 TI - Enzymatic surface-initiated polymerization: a novel approach for the in situ solid-phase synthesis of biocompatible polymer poly(3-hydroxybutyrate). AB - A novel system for surface-initiated enzymatic polymerization of a film of polyhydroxyalkanoate (PHA) on solid surfaces has been developed and characterized. PHAs are aliphatic polyesters produced by a variety of microorganisms as a reserve of carbon and energy, and their properties range from elastomers to thermoplastics, depending on their monomeric composition. The PHA synthase from Ralstonia eutropha H16 was expressed as a poly-histidine fusion in Escherichia coli and immobilized onto several solid substrates through a transition-metal complex, Ni(2+)-nitrilotriacetic acid. The immobilized PHA synthase catalyzed the surface-initiated polymerization of 3-(R)-hydroxybutyryl CoA, forming a polymer film with a uniform thickness on the surface. In this work, we describe the patterned immobilization of the intact enzyme on silicon and subsequent enzymatic polymerization. The immobilized enzyme had a lower specific activity and did not exhibit a lag phase as compared to the soluble enzyme. PMID- 15132679 TI - Synthesis and biological evaluation of a cross-linked hyaluronan-mitomycin C hydrogel. AB - A cross-linked hyaluronan (HA) hydrogel that contained a covalently bound derivative of the anti-proliferative drug mitomycin C (MMC) was synthesized and evaluated in vitro and in vivo. The HA-MMC hydrogel was prepared by coupling MMC aziridinyl-N-acrylate with thiol-modified HA followed by cross-linking with poly(ethylene glycol) diacrylate (PEGDA). MMC was released from 0.5% and 2.0% MMC films by hydrolysis in proportion to the MMC loading. When incubated in vitro with human T31 tracheal scar fibroblasts, 0.5% MMC films inhibited proliferation, whereas 2.0% MMC films were cytotoxic. When implanted in vivo into a rat peritoneal cavity, neither 0.5% nor 2.0% HA-MMC films elicited a severe peritoneal fluid leukocyte response. Importantly, MMC reduced the thickness of fibrous tissue formed surrounding the implanted films. Thus, cross-linked HA-MMC films have strong potential as anti-fibrotic barriers for the prevention of post surgical adhesions. PMID- 15132680 TI - Synthesis, characterization, and bioavailability of mannosylated shell cross linked nanoparticles. AB - Saccharide-functionalized shell cross-linked (SCK) polymer micelles designed as polyvalent nanoscaffolds for selective interactions with receptors on Gram negative bacteria were constructed from mixed micelles composed of poly(acrylic acid-b-methyl acrylate) and mannosylated poly(acrylic acid-b-methyl acrylate). The mannose unit was conjugated to the hydrophilic chain terminus of the amphiphilic diblock copolymer precursor, from which the SCK nanoparticles were derived, by the growth of the diblock copolymer from a mannoside functionalized atom transfer radical polymerization (ATRP) initiator. Mixed micelle formation between the amphiphilic diblock copolymer and mannosylated amphiphilic diblock copolymer was followed by condensation-based cross-linking between the acrylic acid residues present in the periphery of the polymer micelles to afford SCK nanoparticles. SCKs presenting variable numbers of mannose functionalities were prepared from mixed micelles of controlled stoichiometric ratios of mannosylated and nonmannosylated diblock copolymers. The polymer micelles and SCKs were characterized by dynamic light scattering (DLS), electrophoretic light scattering, atomic force microscopy (AFM), transmission electron microscopy (TEM), and analytical ultracentrifugation (AU). Surface availability and bioactivity of the mannose units were evaluated by interactions of the nanostructures with the model lectin Concanavalin A via DLS studies, with red blood cells (rabbit) via agglutination inhibition assays and with bacterial cells (E. coli) via TEM imaging. PMID- 15132681 TI - Microcapsule modification with peroxidase-catalyzed phenol polymerization. AB - A biocatalytic polymer synthesis on a surface of polyelectrolyte microcapsules was studied. Horseradish peroxidase assembled in nanoorganized capsule walls by alternate adsorption with linear polyions retains its activity in reactions of enzyme-catalyzed polymerization of 4-oxyphenols. It allowed controllable synthesis of a phenolic polymer layer on microcapsule walls using an outermost surface peroxidase layer as a template. By varying the phenol type, buffer pH, and reaction component concentrations, the phenolic polymer coating of the capsules with a thickness in the range 20-50 nm was formed. The polymeric products are fluorescent, which provided a good opportunity for confocal image analysis of the capsule wall structure and the attached layer. The influence of a phenolic polymer layer on the permeability of the capsule walls was investigated. PMID- 15132682 TI - Tailoring of bioresorbable polymers for elaboration of sugar-functionalized nanoparticles. AB - Maleic copolymers with different contents of galactose moieties and dodecyl chains were synthesized and used as both a stabilizer and a surface coating for the preparation of poly(epsilon-caprolactone) nanoparticles by the emulsification diffusion technique. The size of the nanoparticles was controlled by varying the initial concentration of the modified maleic copolymers. As the concentration of the latter increased, the particle size decreased, indicating that the copolymers serve as a stabilizer. Moreover, surface modification of nanoparticles was confirmed by xi-potential measurements. Nanoparticles were also shown to be recognized by a galactose-specific lectin, demonstrating the presence of galactose units on the particle surface. This approach offers opportunities for the production of novel targeted drug delivery systems. PMID- 15132683 TI - Structural analysis of chitosan mediated DNA condensation by AFM: influence of chitosan molecular parameters. AB - Chitosan is a nontoxic and biodegradable polysaccharide that has recently emerged as a promising candidate for gene delivery. Here the ability of various chitosans, differing in the fractional content of acetylated units (F(A)) and the degree of polymerization (DP), to compact DNA was studied. Polyplexes made from mixing plasmid DNA with chitosan yielded a blend of toroids and rods, as observed by AFM. The ratios between the fractions of toroids and rods were observed to decrease with increasing F(A) of the chitosan, indicating that the charge density of chitosan, proportional to (1 - F(A)), is important in determining the shape of the compacted DNA. The amount of chitosan required to fully compact DNA into well defined toroidal and rodlike structures were found to be strongly dependent on the chitosan molecular weight, and thus its total charge. A higher charge ratio (+/-) was needed for the shorter chitosans, showing that an increased concentration of the low DP chitosan could compensate for the reduced interaction strength of the individual ligands with DNA. Employing chitosans with different molecular parameters offers the possibility of designing DNA-chitosan polyplexes with various geometries, reflecting various chitosan-DNA interaction strengths, which is necessary for the evaluation of efficient gene delivery vehicles. PMID- 15132684 TI - Surface plasmon resonance study of carbohydrate-carbohydrate interaction between various gangliosides and Gg3-carrying polystyrene. AB - Carbohydrate-carbohydrate interactions between Gg3 trisaccharide-carrying polystyrene (PN(Gg3)) and monolayers of several glycosphingolipids (GSLs) were quantitatively investigated by surface plasmon resonance techniques. PN(Gg3) was adsorbed onto a GM3 monolayer strongly and specifically with an apparent affinity constant of K(a) = 2.5 x 10(6) M(-1), and the apparent affinity constants onto GSLs decreased in the following order: GM3 > LacCer > (KDN)GM3 approximately GlcCer > GM2 approximately GD3 approximately GM4 > GM1 approximately 2,6-isoGM3 > ceramide. These results suggest that PN(Gg3) recognizes not only some specified portions of GM3 but also the trisaccharide as a whole. On the other hand, PN(Lac) and PN(Cel) were bound to GSLs less strongly (K(a) approximately 10(4) M(-1)) and less selectively. The kinetic analysis revealed that the selectivity in the adsorption of PN(Gg3) onto the GM3 monolayer is dominated by the faster adsorption rate. PMID- 15132685 TI - Molecular model for astringency produced by polyphenol/protein interactions. AB - Polyphenols are responsible for the astringency of many beverages and foods. This is thought to be caused by the interaction of polyphenols with basic salivary proline-rich proteins (PRPs). It is widely assumed that the molecular origin of astringency is the precipitation of PRPs following polyphenol binding and the consequent change to the mucous layer in the mouth. Here, we use a variety of biophysical techniques on a simple model system, the binding of beta-casein to epigallocatechin gallate (EGCG). We show that at low EGCG ratios, small soluble polydisperse particles are formed, which aggregate to form larger particles as EGCG is added. There is an initial compaction of the protein as it binds to the polyphenol, but the particle subsequently increases in size as EGCG is added because of the incorporation of EGCG and then to aggregation and precipitation. These results are shown to be compatible with what is known of astringency in foodstuffs. PMID- 15132686 TI - Biomimetic peptide-amphiphiles for functional biomaterials: the role of GRGDSP and PHSRN. AB - The study we present involves the use of a biomimetic system that allows us to study specific interactions in the alpha(5)beta(1) receptor-GRGDSP ligand system with an atomic force microscope (AFM). Bioartificial membranes that mimic the adhesion domain of the extracellular matrix protein fibronectin are constructed from peptide-amphiphiles. A novel peptide-amphiphile is designed that contains both GRGDSP (Gly-Arg-Gly-Asp-Ser-Pro, the primary recognition site for alpha(5)beta(1)) and PHSRN (Pro-His-Ser-Arg-Asn, the synergy binding site for alpha(5)beta(1)) sequences in a single peptide formulation, separated by a spacer. Two different antibodies are used to immobilize and activate isolated alpha(5)beta(1) integrins on the AFM tip. The interaction measured between immobilized alpha(5)beta(1) integrins and peptide-amphiphiles is specific for integrin-peptide binding and is affected by divalent cations in a way that accurately mimics the adhesion function of the alpha(5)beta(1) receptor. The strength of the PHSRN synergistic effect depends on the accessibility of this sequence to alpha(5)beta(1) integrins. An increase in adhesion is observed compared to surfaces displaying only GRGDSP peptides when the new biomimetic peptide-amphiphiles are diluted with lipidated poly(ethylene glycol), which provides more space for the peptide headgroups to bend and expose more of the PHSRN at the interface. PMID- 15132687 TI - Effect of water on the molecular mobility of elastin. AB - Purified and hydrated elastin is studied by both thermal and dielectric techniques to have insight into the chain dynamics of this protein. By differential scanning calorimetry, the glassy behavior of elastin is highlighted; the glass transition temperature (T(g)) of elastin is found to be widely dependent on hydration, falling from 200 degrees C in the dehydrated state to 30 degrees C for 30% hydration. A limit of T(g) at around 0 degrees C is found when crystallizable water is present in the system, that is, when the formation of ice prevents motions of some 10 nm along the polypeptidic chains. The technique of thermally stimulated currents, carried out in the -180 to 0 degrees C temperature range, is useful to detect localized motions. In this case, too, the localized motions vary considerably according to hydration: a first relaxation mode is observed at -145 degrees C and it is associated with the reorientation of crystallizable water in ice I; a second relaxation mode, more complex and cooperative, occurs at around -80 degrees C and could be attributed to the complex constituted by the dipolar groups of the polypeptidic chain and noncrystallizable water, behaving as a glassy system. PMID- 15132688 TI - Stereoselective polymerization of rac-lactide using a monoethylaluminum Schiff base complex. AB - A monoethylaluminum Schiff base complex (2) with formula LAlEt (L = N,N'-(2,2 dimethylpropylene)bis(3,5-di-tert-butylsalicylideneimine) was synthesized and employed for the stereoselective ring-opening polymerization of rac-lactide (rac LA). The complex 2 was characterized by nuclear magnetic resonance, crystal structure, and elemental analysis. It contains a five-coordinate aluminum atom with distorted trigonal bipyramidal geometry in the solid state. In the presence of 2-propanol, 2 showed high stereoselectivity for the polymerization of rac-LA. The polymerization yielded crystalline poly(rac-LA) with a high melting temperature (193-201 degrees C). NMR, differential scanning calorimetry, and wide angle X-ray diffraction indicated that the poly(rac-LA) was highly isotactic, and a stereocomplex was formed between poly-l- and poly-d-lactide block sequences. By the analysis of electrospray-ionization mass spectrometry and (1)H NMR, the polymer was demonstrated to be endcapped in both terminals with an isopropyl ester and a hydroxy group, respectively. The polymerization was of first order in rac-LA concentration. The relationship between the rac-LA conversion and molecular weights of the polymer was linear so that the polymerization could be well controlled. PMID- 15132689 TI - NMR imaging of the diffusion of water at 37 degrees C into Poly(2-hydroxyethyl methacrylate) containing aspirin or vitamin B(12). AB - The ingress of water into poly(2-hydroxyethyl methacrylate), PHEMA, loaded with either one of two model drugs, vitamin B(12) or aspirin, was studied at 37 degrees C using three-dimensional NMR imaging. PHEMA was loaded with 5 and 10 wt % of the drugs. From the imaging profiles, it was observed that incorporation of vitamin B(12) into PHEMA resulted in enhanced crack formation on sorption of water and the crack healing behind the diffusion front was slower than for PHEMA without added drug. This was accounted for by the anti-plasticization of PHEMA by vitamin B(12). Crack formation was inhibited in the PHEMA-aspirin systems because of the plasticizing effect of the aspirin on the PHEMA matrix. All of the polymers were found to absorb water according to an underlying Fickian diffusion mechanism. For PHEMA loaded with 5 wt % of aspirin or vitamin B(12), the best values of the water diffusion coefficients were both found to be 1.3 +/- 0.1 x 10(-11) m(2) s(-1) at 37 degrees C, while the values for the polymer loaded with 10 wt % of the drugs were slightly higher, 1.5 +/- 0.1 x 10(-11) m(2) s(-1). PMID- 15132690 TI - Polymerization of polyfunctional macromolecules: synthesis of a new class of high molecular weight poly(amino acid)s by oxidative coupling of phenol-containing precursor polymers. AB - Oxidative coupling of phenol-containing precursor poly(amino acid)s, poly(alpha glutamine), poly(alpha/beta-asparagine), and poly(gamma-glutamine) derivatives, has been examined to produce a new class of soluble poly(amino acid)s. Under appropriate reaction conditions, the Fe-salen and HRP catalysts efficiently induced the oxidative coupling without formation of insoluble gels, yielding the soluble polymers of high molecular weight. The oxidative coupling behaviors were greatly influenced by the structure and phenol content of the precursor polymer. The selection of the substrate concentration and catalyst amount was crucial for the production of soluble polymers of high molecular weight. PMID- 15132691 TI - Effect of surfactant type on surfactant--protein interactions at the air-water interface. AB - The displacement of the proteins (beta-lactoglobulin and beta-casein) from an air water interface by the nonionic (Tween 20 and Tween 60) and ionic (sodium dodecyl sulfate, cetyltrimethylammonium bromide, and lyso-phosphatidylcholine-lauroyl) surfactants has been visualized by atomic force microscopy (AFM). The surface structure has been sampled by the use of Langmuir-Blodgett deposition onto mica substrates to allow imaging in the AFM. In all cases, the displacement process was found to occur through the recently proposed orogenic mechanism (Mackie et al. J. Colloid Interface Sci. 1999, 210, 157-166). In the case of the nonionic surfactants, the displacement involved nucleation and growth of surfactant domains leading to failure of the protein network and subsequent loss of protein into the bulk phase. The surface pressure dependence of the growth of surfactant domains and the failure of the network were found to be the same for both Tween 20 and Tween 60, demonstrating that the breakdown of the protein film was dominated by the mechanical properties of the network. The displacement of protein by ionic surfactants was found to be characterized by nucleation of surfactant domains with little domain growth prior to failure of the network. The size of the domains formed by ionic surfactants was found to be limited by the strong intersurfactant repulsive forces between the charged headgroups. Screening of these charges led to an increase in the size of the domains. The surface pressure at which the network continuity was lost was found to be dependent on the type of surfactant and, in all cases, to occur at higher surface pressures than that required for nonionic surfactants. This has been attributed to surfactant-protein binding that initially strengthens the protein network at low surfactant concentrations. Evidence obtained from surface shear rheology supports this assertion. PMID- 15132692 TI - Comparative study of the first heterogeneous deacetylation of alpha- and beta chitins in a multistep process. AB - Heterogeneous deacetylation of alpha- and beta-chitins from shrimp shells and squid pens were comparatively studied. Each deacetylated sample, recovered after neutralization, was fractionated according to a water-soluble and insoluble fraction (pH 8.5). The systematic study of DAs, crystallinity changes, and distribution of N-acetylglucosamine residues were performed on the two kinds of fractions. For the two fractions resulting from a deacetylation in the presence of 50% (w/v) NaOH, for temperatures ranging from 80 to 110 degrees C, the activation energies of the reactions were found close to 39.9 +/- 1.0 and 42.8 +/ 1.8 kJ mol(-1) and the frequency factors of collision were of 7.2 +/- 2.4 and 54.4 +/- 18.5 10(3) min(-1), for alpha- and beta-chitins, respectively. Deacetylations of water-soluble and insoluble fractions were compared, and the major role played by the crystallinity level during deacetylation was evidenced and, thereafter, the role of the nature of the starting chitins on the chemical behavior. Thus, in some conditions, we observed critical values of DDA where the structures were becoming fully amorphous. PMID- 15132693 TI - Swelling behavior and structural characteristics of wheat gluten polypeptide films. AB - Wheat gluten films were subjected to controlled thermomechanical treatments to increase the percentage of aggregated sodium dodecyl sulfate (SDS)-insoluble gluten protein, the aggregation reaction being disulfide bonding. The rheological properties of the films were measured under immersion in water, where wheat gluten films are stable and show only slight swelling. The equilibrium swelling of the gluten films in water decreased with the increase of the percentage of SDS insoluble protein aggregates, and the frequency the independent shear modulus increased sharply with increasing percentage of SDS-insoluble aggregates. Both findings confirm that disulfide bonding between gluten proteins is the predominant cross-linking reaction in the system. A relationship between shear modulus and aggregated protein compatible with a power law (of exponent 3) suggests the existence of a protein network at a molecular scale. However, the classical Flory-Rehner model failed to describe the relationship between the plateau modulus and the gluten volume fraction (a very drastic increase, compatible with a power law of an exponent of about 14). This result shows that gluten cannot be described as an entangled polymer network. The interpretation of both relationships is a network of mesoscale particles which in turn have a fractal inner structure (with a fractal dimension close to 3). PMID- 15132694 TI - Novel water-soluble photosensitizers from dextrans. AB - Novel water-soluble polymeric photosensitizers based on the natural polymer dextran were synthesized and studied. The modified dextran contained photoactive anthracene (An) chromophores. They were soluble in water with the solubility decreasing with an increase in the number of An moieties bound to the polymeric chain. In aqueous solutions, the macromolecules adopted a compact conformation which resulted in the formation of hydrophobic microdomains. The properties of these domains were characterized with molecular probes such as perylene and pyrazolo-quinoline derivative. The polymer absorbed in the UV/vis region and photosensitized reactions mediated by energy and/or electron transfer from electronically excited An to the molecules of organic compounds solubilized in polymeric microdomains or resided in water. PMID- 15132695 TI - Dynamics of the Escherichia coli O91 O-antigen polysaccharide in solution as studied by carbon-13 NMR relaxation. AB - The dynamics of the O-antigen part of the lipopolysaccharide from the enterohemorrhagic Escherichia coli O91 has been determined in solution using (13)C NMR relaxation measurements at two magnetic field strengths, 9.4 and 14.1 T, thereby facilitating the testing of several dynamical models. The biological repeating unit, consisting of five sugar residues and substituents, could be determined by spectral analysis of different (1)H,(13)C correlations and corroborated by the relaxation data. The site specifically (13)C-labeled material was shown to have approximately 10 repeating units with a narrow distribution. A model-free analysis of the relaxation data revealed a complex dynamical behavior where the sugar residues could be described by a global correlation time (tau(m) = 5.4 ns), generalized order parameters (S(2) approximately 0.63), and different correlation times for internal motions related to their position in the repeating unit along the polymer (tau(e) approximately 360-520 ps). One of the sugar residues showed, in addition, a chemical exchange contribution. Furthermore, a substituent on another sugar residue was described by two order parameters (S(f)(2) = 0.51 and S(s)(2) = 0.21). The solution dynamics of the polysaccharide are thus described by highly intricate motions, both in amplitude and time scales. These results are of significance in the general description of polysaccharides surrounding bacterial cell surfaces and in the presentation of antigenic epitopes to the immune system of an invaded host. PMID- 15132696 TI - In vitro hydrolysis of poly(l-lactide) crystalline residues as extended-chain crystallites: II. Effects of hydrolysis temperature. AB - The effects of hydrolysis temperature on the hydrolysis behavior and mechanism of poly(l-lactide) crystalline residues or extended-chain crystallites were investigated in phosphate-buffered solution (50-97 degrees C), using gel permeation chromatography and differential scanning calorimetry (DSC). The hydrolysis of the crystalline residues proceeded from their surface composed of very short chains with a free end along the chain direction, irrespective of hydrolysis temperature, but the hydrolysis from their lateral surface could not be traced. The activation energy of hydrolysis for the crystalline residues (extended-chain crystallites) was evaluated to be 18.0 kcal mol(-1) (75.2 kJ mol( 1)). The monotonic melting temperature (T(m)) and crystallinity decreases occurred after their initial very small increases, excluding the monotonic crystallinity decrease at 97 degrees C with no initial increase. The T(m) decrease reflects the decreased thickness of the crystalline residues. The equilibrium T(m) of the crystalline residues (extended-chain crystallites) was estimated to be 464.5-464.9 K. The free energy values for the surface composed of very short chains with a free end, which are neighboring the air (or nitrogen during DSC scanning), were calculated to be 55.6-56.4 erg cm(-2) for heat of fusion per unit mass = 135 J g(-1). The obtained surface free energy values are significantly higher than that for the surface composed of folding chains, tie chains, and the chains with a free end, which are neighboring the same kind of amorphous chains (39.9 erg cm(-2)). PMID- 15132697 TI - Accelerated biodegradation of poly(vinyl alcohol) by glycosidations of the hydroxyl groups or addition of sugars. AB - Biodegradabilities of N-acetyl-d-glucosamine (GlcNAc)- (1) and chitobiose substituted (2) poly(vinyl alcohol)s (PVA)s in a soil suspension (pH 6.5) were investigated at 25 degrees C for 40 days. Biochemical oxygen demand of 1 with a degree of substitution of 0.2-0.3 (DP = 430-480) was higher than that of PVA under the degradation condition. Size exclusion chromatography, (1)H NMR, and Fourier-transform infrared measurements of the recovered sample indicated that biodegradation of the PVA main chain was accelerated by partial glycosidation of hydroxyl groups in PVA. Similar acceleration was observed in a PVA/GlcNAc (50:50, w/w) mixture. Microbes which relate with degradation of the glycosidated polymers were grown in a culture medium including the soil suspension and the polymer as the carbon source. Polyacrylamide gel electrophoresis (SDS-PAGE) and IR measurements indicated that a cell-free extract derived from GlcNAc-substituted PVA was different from that in the PVA/GlcNAc mixture. The results suggested that the PVA main chain in GlcNAc-substituted PVA was cleaved by a different microorganism or via a mechanism different from that in the mixture. Chitobiose substituted PVA 2 showed more enhanced acceleration, indicating that the sugar length influenced the degradability. PMID- 15132698 TI - Glucose-responsive polymer gel bearing phenylborate derivative as a glucose sensing moiety operating at the physiological pH. AB - The work attempts to prepare a totally synthetic, glucose-responsive polymer gel bearing a phenylborate derivative as a sensor moiety to glucose, for future use as a self-regulated insulin delivery system. The molecular strategies to enable the system to be operated under physiological conditions (pH 7.4, 37 degrees C) are presented that involve the use of a novel phenylborate derivative [4-(1,6 dioxo-2,5-diaza-7-oxamyl) phenylboronic acid: DDOPBA] possessing an appreciably low pK(a) ( approximately 7.8), the adoption of poly(N-isopropylmethacrylamide) (PNIPMAAm) for the main chain, which itself undergoes a sharp thermo-induced phase transition at its LCST around 40 degrees C, as well as the introduction of a carboxyl group of methacrylic acid as the third comonomer. Glucose-responsive behaviors of the obtained gels were evaluated based on the changes in the equilibrium swelling degree determined in the presence and the absence of glucose, for various pH and temperature conditions. As a consequence of the combined molecular effects, a sufficient sensitivity of the system was accomplished at physiological pH and in the temperature range close to the physiological condition such as 30 degrees C. Furthermore, the glucose-induced continuous volume changes of the gels were demonstrated under those conditions, which occurred in a remarkably concentration-dependent manner. In these experiments, the critical glucose concentrations to induce the gels' responses in the range of normoglycemic sugar level were observed. These observations may provide us with an excellent prospect for the use of the gel as a self-regulated, insulin-delivery system discretely switching the release at the normoglycemia. PMID- 15132699 TI - Morphology and properties of soy protein isolate thermoplastics reinforced with chitin whiskers. AB - Environmentally friendly thermoplastic nanocomposites were successfully developed using a colloidal suspension of chitin whiskers as a filler to reinforce soy protein isolate (SPI) plastics. The chitin whiskers, having lengths of 500 +/- 50 nm and diameters of 50 +/- 10 nm on average, were prepared from commercial chitin by acid hydrolysis. The dependence of morphology and properties on the chitin whiskers content in the range from 0 to 30 wt % for the glycerol plasticized SPI nanocomposites was investigated by dynamic mechanical thermal analysis, scanning electron microscopy, swelling experiment, and tensile testing. The results indicate that the strong interactions between fillers and between the filler and SPI matrix play an important role in reinforcing the composites without interfering with their biodegradability. The SPI/chitin whisker nanocomposites at 43% relative humidity increased in both tensile strength and Young's modulus from 3.3 MPa for the SPI sheet to 8.4 MPa and from 26 MPa for the SPI sheet to 158 MPa, respectively. Further, incorporating chitin whisker into the SPI matrix leads to an improvement in water resistance for the SPI based nanocomposites. PMID- 15132700 TI - Construction, molecular modeling, and simulation of Mycobacterium tuberculosis cell walls. AB - The mycobacterial cell wall is extraordinarily thick and tight consisting mainly of (1). long chain fatty acids, the mycolic acids, and (2). a unique polysaccharide, arabinogalactan (AG). These two chemical constituents are covalently linked through ester bonds. Minnikin (The Biology of the Mycobacteria; Academic: London, 1982) proposed that the mycobacterial cell wall is composed of an asymmetric lipid bilayer. The inner leaflet of the cell wall contains mycolic acids covalently linked to AG. This inner leaflet is believed to have the lowest permeability to organic compounds of the overall cell wall. Conformational search and molecular dynamics simulation were used to explore the conformational profile of AG and the conformations and structural organization of the mycolic acid-AG complex, and overall, an inner leaflet molecular model of the cell wall was constructed. The terminal arabinose residues of AG that serve as linkers between AG and mycolic acids were found to exist in four major chemical configurations. The mycolate hydrocarbon chains were determined to be tightly packed and perpendicular to the "plane" formed by the oxygen atoms of the 5-hydroxyl groups of the terminal arabinose residues. For Mycobacterium tuberculosis, the average packing distance between mycolic acids is estimated to be approximately 7.3 A. Thus, Minnikin's model is supported by this computational study. Overall, this modeling and simulation approach provides a way to probe the mechanism of low permeability of the cell wall and the intrinsic drug resistance of M. tuberculosis. In addition, monolayer models were built for both dipalmitoylphosphatidylethanolamine and dimyristoylphosphatidylcholine, two common phospholipids in bacterial and animal membranes, respectively. Structural comparisons of these cell wall phospholipid membrane models were made to the M. tuberculosis cell wall model. PMID- 15132701 TI - Molecular modeling and simulation of Mycobacterium tuberculosis cell wall permeability. AB - The low permeability of the mycobacterial cell wall is thought to contribute to the intrinsic drug resistance of mycobacteria. In this study, the permeability of the Mycobacterium tuberculosis cell wall is studied by computer simulation. Thirteen known drugs with diverse chemical structures were modeled as solutes undergoing transport across a model for the M. tuberculosis cell wall. The properties of the solute-membrane complexes were investigated by means of molecular dynamics simulation, especially the diffusion coefficients of the solute molecules inside the cell wall. The molecular shape of the solute was found to be an important factor for permeation through the M. tuberculosis cell wall. Predominant lateral diffusion within, as opposed to transverse diffusion across, the membrane/cell wall system was observed for some solutes. The extent of lateral diffusion relative to transverse diffusion of a solute within a biological cell membrane may be an important finding with respect to absorption distribution, metabolism, elimination, and toxicity properties of drug candidates. Molecular similarity measures among the solutes were computed, and the results suggest that compounds having high molecular similarity will display similar transport behavior in a common membrane/cell wall environment. In addition, the diffusion coefficients of the solute molecules across the M. tuberculosis cell wall model were compared to those across the monolayers of dipalmitoylphosphatidylethanolamine and dimyristoylphosphatidylcholine, are two common phospholipids in bacterial and animal membranes. The differences among these three groups of diffusion coefficients were observed and analyzed. PMID- 15132702 TI - Photoinduced graft-copolymer synthesis and characterization of methacrylic acid onto natural biodegradable lignocellulose fiber. AB - UV radiation induced graft copolymerization of methacrylic acid onto natural lignocellulose (jute) fiber was carried out both by "simultaneous irradiation and grafting" and by preirradiation methods using 1-hydroxycyclohexyl-phenyl ketone as a photoinitiator. In the "simultaneous irradiation and grafting" method, the variation of graft weight with UV-radiation time, monomer concentration, and the concentration of photoinitiator was investigated. In the case of the preirradiation method, the incorporation of 2-methyl-2-propene 1-sulfonic acid, sodium salt, into the grafting reaction solution played a most important role in suppressing the homopolymer/gel formation and facilitating graft copolymerization. The optimum value of the reaction parameters on the percentage of grafting was evaluated. In comparison, results showed that the method of graft copolymer synthesis has significant influence on graft weight. The study on the mechanical and thermal properties of grafted samples was conducted. The results showed that the percentage of grafting has a significant effect on the mechanical and thermal properties in the case of grafted samples. Considering the water absorption property, the jute-poly(methacrylic acid)-grafted sample showed a maximum up to 42% increase in hydrophilicity with respect to that of the "as received" sample. Attenuated total reflection infrared studies indicate that the estimation of the degree of grafting could be achieved by correlating band intensities with the percent graft weight. PMID- 15132703 TI - Protein adsorption modalities on polyelectrolyte multilayers. AB - Protein adsorption on polyelectrolyte multilayers (PEMUs) was evaluated using a combination of synthetic polyelectrolytes and proteins, including serum albumin, fibrinogen, and lysozyme. Variables such as surface and protein charge, polymer hydrophobicity, and hydrophilic repulsion were introduced to probe interaction mechanisms. Quantitative analysis with reflectance Fourier transform infrared spectroscopy, optical waveguiding, and UV-vis absorption, together with qualitative information from atomic force microscopy, provided a coordinated picture for what drives protein adsorption and how the molecules are disposed on the multilayer surface. It was found that multilayers bearing a particular surface charge sorbed biomolecules if they were of opposite charge, yielding significant loadings within the bulk PEMU. Adsorption of like-charged proteins, as surface aggregates, occurred to a much lower extent, driven by nonelectrostatic forces. A diblock copolymer comprising a hydrophilic poly(ethylene oxide) block was capable of further minimizing protein adsorption as a result of hydrophilic repulsion, although none of the surfaces tested defeated protein adsorption completely. However, poly(acrylic acid) homopolymer was quite effective in this respect. A composition gradient, formed during multilayer buildup, induced a gradient in hydrophilicity through the PEMU, which is an efficient and economical method of creating a protein-resistant surface. PMID- 15132704 TI - Silica nanocasts of wood fibers: a study of cell-wall accessibility and structure. AB - The porosity and the available surface area of a lignocellulosic fiber can influence the accessibility and reactivity in derivatization and modification reactions because the porous cell-wall network determines the upper size limit for molecules that can penetrate and react with the interior of the wall. To obtain information concerning the accessibility of the porous cell wall of wood fibers, surfactant-templated sol-gel mineralization has been examined. Wood and kraft pulp samples of Norway spruce were impregnated with a silica sol-gel and subsequently heated (calcined) and transformed into structured mesoporous silica. Microscopy studies (environmental scanning electron microscopy, transmission electron microsopy, TEM) on the silica casts showed that the three-dimensional architecture of the wood and pulp fiber cell wall was revealed down to the nanometer level. Image analysis of TEM micrographs of silica fragments from the never-dried pulp revealed complete infiltration of the cell-wall voids and microcavities (mean pore width 4.7 +/- 2 nm) by the sol-gel and the presence of cellulose fibrils with a width of 3.6 +/- 1 nm. Cellulose fibrils of the same width as that shown by image analysis were also identified by nitrogen adsorption measurements of the pore size distribution in the replicas. PMID- 15132705 TI - Poly(amidoamine) salt form: effect on pH-dependent membrane activity and polymer conformation in solution. AB - On exposure to an acidic pH, linear poly(amidoamine)s (PAAs) cause membrane perturbation and consequently have potential as endosomolytic polymers for the intracellular delivery of genes and toxins. Previous studies used PAAs in the hydrochloride form only. The aim of this study was to investigate systematically the effect of the PAA counterion on pH-dependent membrane activity, general cytotoxicity, and PAA solution properties to help guide optimization of PAA structure for further development of PAA-protein conjugates. PAAs (ISA 1, 4, 22, and 23; M(w) 10000-50000 g/mol) were synthesized to provide a library of PAAs having different counterions including the acetate, citrate, hydrochloride, lactate, phosphate, and sulfate salts. pH-Dependent membrane activity was assessed using a rat red blood cell haemolysis assay (conducted at a starting pH of 7.4, 6.5, or 5.5; 1 mg/mL; 1 h), and general cytotoxicity was investigated using a murine melanoma cell line (B16F10) and a human bladder endothelial-like cell line (ECV-304). Whereas poly(ethyleneimine) was haemolytic at the starting pH of 7.4 at 1 h [ approximately 50% haemoglobin (Hb) release], none of the PAA salts were haemolytic at a starting pH of 7.4 or 6.5. Although PAA acetate, citrate, and lactate were also non-haemolytic at the starting pH of 5.5, the sulfate and hydrochloride forms caused significant haemolysis (up to 80% Hb release) and ISA 22 and 23 phosphate were also markedly haemolytic ( approximately 70% Hb release). These counterion-specific differences were also clearly visible using scanning electron microscopy, which was used to visualize the red blood cell morphology. All PAAs were relatively nontoxic (IC(50) >or= 300 5000 microg/mL) compared to poly-l-lysine (IC(50) = 2-10 microg/mL), the PAA hydrochloride salts produced the greatest cytotoxicity, and the B16F10 cells were more sensitive than the ECV-304 cells. Small-angle neutron scattering suggested that ISA 23 hydrochloride had a larger hydrodynamic radius (5.1 +/- 0.2 nm) than the citrate salt (3.1 +/- 0.2 nm). These results provide indirect evidence for the salt- and pH-dependent changes in the conformation of the polymer coil. This study clearly demonstrates the importance of optimization of the counterion form when developing endosomolytic polymers designed to mediate pH-dependent membrane permeabilization. PMID- 15132706 TI - Novel biodegradable polyphosphate cross-linker for making biocompatible hydrogel. AB - To obtain a novel biodegradable cross-linker, polymerizable polyphosphate (PIOP) was synthesized by ring-opening polymerization of 2-i-propyl-2-oxo-1,3,2 dioxaphospholane with 2-(2-oxo-1,3,2-dioxaphosphoroyloxyethyl methacrylate) (OPEMA). The number averaged molecular weight of the PIOP was 1.2 x 10(4), and the number of OPEMA units in one PIOP molecule was 2.2. Nonenzymatic degradation of the PIOP was evaluated in various pH aqueous media. The degree of hydrolysis was dependent on the pH; that is, it increased with an increase in the pH of the medium. At pH 11.0, the PIOP completely degraded in only 6 days. The poly[2 methacryloyloxyethyl phosphorylcholine (MPC)] cross-linked with the PIOP was prepared by radical polymerization. This polymer could form hydrogel, and the free water fraction in the hydrogel was high. The enzymatic activity of trypsin in contact with the hydrogel was similar to that in buffer solution. There is no adverse effect caused by the hydrogel to reduce the function of the trypsin. The cytotoxicity of poly(MPC) and degraded PIOP was evaluated using v79 cells, and it was not observed in either case. In conclusion, PIOP is a hydrolyzable polymer, which can be used as a cross-linker, and novel hydrogels having biodegradability and biocompatibility were prepared from poly(MPC) cross-linked with the PIOP. PMID- 15132707 TI - Effects of the molecular structure of two amphiphilic antidepressant drugs on the formation of complexes with human serum albumin. AB - Interactions of two amphiphilic antidepressant drugs, imipramine and desipramine hydrochlorides, with the blood protein human serum albumin (HSA) were investigated to gain an understanding of the effects of drug molecular structure on the complex formation of drug-protein molecules. To elucidate the mechanisms of such effects, the protein-antidepressant interactions in aqueous buffered solutions of pH 3.0 and 5.5 (isoelectric point of HSA = 4.9) were investigated using conductivity, zeta potential, and dynamic light scattering. An increase of the critical micelle concentration of both antidepressants was detected as a consequence of extensive binding to the protein. From zeta-potential measurements, the Gibbs energies of adsorption of the drugs onto the protein were derived using the proposed models of Kayes and Ottewill and Watanabe. Measurements of the hydrodynamic radii of HSA-antidepressant complexes as a function of the drug concentration have shown a gradual increase of size of a saturation rather than a denaturation process of the protein. A larger drug adsorption at pH 5.5 than at pH 3.0 was also observed, as a consequence of a more important specific binding at the former pH. PMID- 15132708 TI - Biodegradable films of partly branched poly(l-lactide)-co-poly(epsilon caprolactone) copolymer: modulation of phase morphology, plasticization properties and thermal depolymerization. AB - We report on the modulation of phase morphology, plasticization properties, and thermal stability of films of partly branched poly(l-lactide)-co-poly(epsilon caprolactone) copolymer (PLLA-co-PCL) with additions of low molecular weight compounds, namely, triethyl citrate ester, diethyl phthalate, diepoxy polyether (poly(propylene glycol) diglycidyl ether), and with epoxidized soybean oil (ESO). The PLLA-co-PCL/polyether films showed significant stability against thermal depolymerization, high film flexibility, and good plasticizing properties, probably due to cross-linking and chain branching formation between diepoxy groups with both the end carboxyl and hydroxyl groups of the PLLA copolymer (initially present or generated during the degradation process) to produce primary ester and ether bonds, respectively. Diethyl phthalate and triethyl citrate ester were found to be efficient plasticizers for PLLA copolymer in terms of glass transition and mechanical properties, but the more water-soluble plasticizer triethyl citrate induced a dramatic loss in the molecular weight of the copolymer. Although ESO cannot play the role of a plasticizer, it substantially stabilizes and retards thermal depolymerization of the PLLA copolymer matrix, possibly because of a reaction between epoxy groups with the end carboxyl and hydroxyl groups of the PLLA copolymer. The presence of ESO in PLLA-co-PCL/ESO/triethyl citrate blends enhanced the compatibility and miscibility of the plasticizer with the PLLA copolymer matrix, considerably improved the mechanical properties (elongation at break), and substantially stabilized the copolymer against thermal depolymerization. It seems likely that the epoxy groups interact not only with the end hydroxyl and carboxyl group of the copolymer but as well with the hydroxyl group of triethyl citrate plasticizer to produce a new ether bond (C-O-C) as the cross-linking unit. On the other hand, for PLLA-co-PCL/ESO/polyether blends, (80/10/10) epoxidized oil distorts the compactness of the blend by diminishing the proposed entanglements between carboxyl, hydroxyl, and diepoxy groups of polyether and reduces the high elongation properties otherwise observed in the PLLA-co-PCL/polyether films. The multicomponent approach toward modulating poly(l-lactide)-co-poly(epsilon caprolactone) copolymer films using epoxy compounds and plasticizers and the insight into the nature of various PLLA matrixes presented here offer advantages to a broad engineering of PLLA copolymer films having desirable physical properties and multiphase behavior for efficient uses in future technical applications. PMID- 15132709 TI - Modal difference in comonomer-unit compositional distributions of poly(3 hydroxybutyrate-co-4-hydroxybutyrate)s biosynthesized by two strains, Ralstonia eutropha and Alcaligenes latus. PMID- 15132710 TI - Treatment strategies in Alzheimer's disease with a focus on early pharmacological interventions. AB - Complex interactive effects of genetic predisposition, neurochemical changes and disease comorbidity have been elucidated in the genesis of dementia syndromes. Alzheimer's disease is the most prevalent type of dementia in developed Western countries. In Alzheimer's disease, pharmacological treatment aims at symptomatic relief, disease modification or disease prevention. Cholinesterase inhibitors are established for the treatment of mild-to-moderate Alzheimer's disease. In Europe and the US, memantine is approved for the treatment of moderate-to-severe Alzheimer's disease. To date, there are no drugs with a disease modifying action that have proven efficacy in randomised, double-blind, placebo-controlled clinical trials. In patients not fulfilling the diagnostic criteria for early Alzheimer's disease, e.g. mild cognitive impairment, the efficacy of several drugs, mainly cholinesterase inhibitors, is currently tested in prospective studies by determining the conversion rate to Alzheimer's disease. However, prevention and disease-modifying strategies raise ethical questions because interventions are focused on non-diseased elderly at risk, which means that emphasis should be not only on efficacy but also on long-term safety. No disease modifying strategy can presently be offered to patients; however, given the pace of recent research there is optimism that slowing progression of Alzheimer's disease will soon be possible. PMID- 15132711 TI - Life-threatening idiosyncratic drug-induced agranulocytosis in elderly patients. AB - Agranulocytosis is a life-threatening disorder in any age, but particularly so in elderly patients who are receiving, on average, a larger number of drugs than younger patients. Drug-induced agranulocytosis still remains a rare event, with an annual incidence rate of approximately 3-12 cases per million population. This disorder frequently occurs as an adverse reaction to drugs, particularly antibacterials, antiplatelet agents, antithyroid drugs, antipsychotics or antiepileptic drugs, and NSAIDs. Although patients experiencing drug-induced agranulocytosis may initially be asymptomatic, the severity of the neutropenia usually translates into the onset of severe sepsis that requires intravenous broad-spectrum antibacterial therapy. In this setting, haematopoietic growth factors have been shown to shorten the duration of neutropenia. Thus, with appropriate management, the mortality rate of idiosyncratic drug-induced agranulocytosis is now 5-10%. However, given the increased life expectancy and subsequent longer exposure to drugs, as well as the development of new agents, physicians should be aware of this complication and its management. PMID- 15132712 TI - Adjuvant therapy for rectal cancer in the elderly. AB - Rectal cancer, like most malignancies, is a disease of older age. By the year 2030, nearly 70% of all cancer patients are expected to be over the age of 65 years. Adjuvant therapy for colorectal cancer has been one of the most important contributions of medical oncology to the health of the population, saving more lives annually than more effective therapy for less common cancers, such as Hodgkin's disease. Nonetheless, population-based studies have shown that less than half of those over the age of 65 years receive the standard adjuvant therapy for rectal cancer. In many instances, there may be legitimate reasons for this, but efforts must be made to overcome any age bias and nihilism in the use of adjuvant therapy in the treatment of rectal cancer in the elderly. Although the elderly have been under-represented in clinical trials, they have been shown to tolerate cancer treatment and derive benefit from the adjuvant therapies. Despite early reports to the contrary, older patients have been shown to tolerate surgical resections for rectal cancer as well as their younger counterparts. Studies have supported the use of combined modality therapy as standard adjuvant care for clinical T3 rectal cancer in the preoperative setting and for patients with T3 and/or N1/N2 disease in the postoperative setting, wtih improved rates of sphincter preservation, recurrence and overall survival. PMID- 15132714 TI - Spontaneous reports of hypertension leading to hospitalisation in association with rofecoxib, celecoxib, nabumetone and oxaprozin. AB - BACKGROUND AND OBJECTIVE: Data on file with the US FDA, and other published studies, suggest that the selective cyclo-oxygenase (COX)-2 inhibitor NSAID rofecoxib has a greater hypertensive adverse effect than other NSAIDs, including celecoxib. In this study we describe a pharmacoepidemiologic analysis of spontaneous adverse event reports of acute, clinically serious hypertension (as defined by hospitalisation) reported in association with rofecoxib, celecoxib, nabumetone and oxaprozin. The objective of this analysis is to assess whether postmarketing data are consistent with results of clinical trials. We also collapse cases into series for the identification of possible risk factors for clinically severe, NSAID-associated hypertension. METHODS: Domestic (US) cases of apparently unconfounded, acute hypertension leading to hospitalisation were collected and reviewed from the spontaneous adverse events database of the FDA for rofecoxib, celecoxib, nabumetone and oxaprozin for the initial 3 years of marketing. Drug use data for the same intervals enabled calculation of reporting rates. RESULTS: In an analysis of reporting rates, hospitalisation for acute blood pressure (BP) elevation was reported more frequently (3.8-fold) for rofecoxib compared with celecoxib. A total of 34 cases are collapsed into case series. No cases were identified for either nabumetone or oxaprozin. Inspection of reviewed cases for celecoxib and rofecoxib suggest that these patients (average age 72 years) were potentially high-risk candidates for NSAID therapy. DISCUSSION AND CONCLUSION: During early marketing, hospitalisation for acute BP elevation appears to have been reported more frequently for rofecoxib compared with celecoxib. This is consistent with clinical trial data on file with the FDA, and other published studies that found rofecoxib to have a greater effect on BP than other NSAIDs, including celecoxib. This finding may be particularly relevant in older patients given the prevalence of hypertension and cardiovascular disease in this age group. PMID- 15132715 TI - The environment and autoimmune thyroid diseases. AB - Genetic factors play an important role in the pathogenesis of autoimmune thyroid disease (AITD) and it has been calculated that 80% of the susceptibility to develop Graves' disease is attributable to genes. The concordance rate for AITD among monozygotic twins is, however, well below 1 and environmental factors thus must play an important role. We have attempted to carry out a comprehensive review of all the environmental and hormonal risk factors thought to bring about AITD in genetically predisposed individuals. Low birth weight, iodine excess and deficiency, selenium deficiency, parity, oral contraceptive use, reproductive span, fetal microchimerism, stress, seasonal variation, allergy, smoking, radiation damage to the thyroid gland, viral and bacterial infections all play a role in the development of autoimmune thyroid disorders. The use of certain drugs (lithium, interferon-alpha, Campath-1H) also increases the risk of the development of autoimmunity against the thyroid gland. Further research is warranted into the importance of fetal microchimerism and of viral infections capable of mounting an endogenous interferon-alpha response. PMID- 15132716 TI - The emerging role of the CTLA-4 gene in autoimmune endocrinopathies. AB - It is thought that the majority of autoimmune endocrinopathies, including Graves' disease, autoimmune hypothyroidism, type 1 diabetes mellitus and autoimmune Addison's disease (sporadic and as well as autoimmune polyendocrinopathy syndrome type 2) are inherited as complex genetic traits. Multiple genetic and environmental factors interact with each other to confer susceptibility to these disorders. In recent years there have been considerable efforts towards defining susceptibility genes for complex traits. These investigations have shown, with increasing evidence, that the cytotoxic T lymphocyte antigen-4 (CTLA-4) gene is an important susceptibility locus for autoimmune endocrinopathies and other autoimmune disorders. Here we review the genetic and functional analyses of the CTLA-4 locus in autoimmune endocrinopathies, and discuss the recent efforts in fine-mapping this locus. PMID- 15132717 TI - GLUT1 deficiency and other glucose transporter diseases. AB - We review the three genetically determined disorders of glucose transport across cell membranes. Diseases such as glucose-galactose malabsorption, Fanconi-Bickel syndrome and De Vivo disease (GLUT1 deficiency syndrome (GLUT1DS)) arise from heritable mutations in transporter-encoding genes that impair monosaccharide uptake, which becomes rate-limiting in tissues where the transporters serve as the main glucose carrier systems. We focus in greater detail on De Vivo disease as a prototype of a brain energy failure syndrome, for which the greatest pathophysiological detail is known, but which presents the most therapeutic challenges. The study of these diseases illustrates fundamental aspects of energetic metabolism, while providing the basis for their diagnosis by simple metabolic screening and for their treatment by dietary modification. PMID- 15132713 TI - Cholinesterase inhibitors used in the treatment of Alzheimer's disease: the relationship between pharmacological effects and clinical efficacy. AB - The deficiency in cholinergic neurotransmission in Alzheimer's disease has led to the development of cholinesterase inhibitors as the first-line treatment for symptoms of this disease. The clinical benefits of these agents include improvements, stabilisation or less than expected decline in cognition, function and behaviour. The common mechanism of action underlying this class of agents is an increase in available acetylcholine through inhibition of the catabolic enzyme, acetylcholinesterase. There is substantial evidence that the cholinesterase inhibitors, including donepezil, galantamine and rivastigmine, decrease acetylcholinesterase activity in a number of brain regions in patients with Alzheimer's disease. There is also a significant correlation between acetylcholinesterase inhibition and observed cognitive improvement. However, the cholinesterase inhibitors are reported to have additional pharmacological actions. Rivastigmine inhibits butyrylcholinesterase with a similar affinity to acetylcholinesterase, although it is not clear whether the inhibition of butyrylcholinesterase contributes to the therapeutic effect of rivastigmine. Based on data from preclinical studies, it has been proposed that galantamine also potentiates the action of acetylcholine on nicotinic receptors via allosteric modulation; however, the effects appear to be highly dependent on the concentrations of agonist and galantamine. It is not yet clear whether these concentrations are related to those achieved in the brain of patients with Alzheimer's disease within therapeutic dose ranges. Preclinical studies have shown that donepezil and galantamine also significantly increase nicotinic receptor density, and increased receptor density may be associated with enhanced synaptic strengthening through long-term potentiation, which is related to cognitive function. Despite these differences in pharmacology, a review of clinical data, including head-to-head studies, has not demonstrated differences in efficacy, although they may have an impact on tolerability. It seems clear that whatever the subsidiary modes of action, clinical evidence supporting acetylcholinesterase inhibition as the mechanism by which cholinesterase inhibitors treat the symptoms of Alzheimer's disease is accumulating. Certainly, as a class, the currently approved cholinesterase inhibitors (donepezil, galantamine, rivastigmine and tacrine) provide important benefits in patients with Alzheimer's disease and these drugs offer a significant advance in the management of dementia. PMID- 15132718 TI - Primary GH insensitivity '(Laron syndrome) caused by a novel 4 kb deletion encompassing exon 5 of the GH receptor gene: effect of intermittent long-term treatment with recombinant human IGF-I. AB - OBJECTIVE: GH insensitivity syndrome (GHIS; Laron syndrome) is clinically characterized by severe postnatal growth failure and very low serum levels of IGF I despite increased secretion of GH. This mainly autosomal recessive syndrome is clinically indistinguishable from isolated GH deficiency (IGHD). Fifty-one different mutations in the GH receptor (GHR) gene have been discovered, whereas only three deletions causing the disorder have been reported so far. In this report, we describe a consanguineous family from Sri Lanka with a novel deletion of 4097 bp in length encompassing exon 5. SUBJECTS AND METHODS: Parents of normal phenotype presented their second child (boy) to our clinic at the age of 7 months with severe growth retardation and the clinical features of IGHD (58 cm, -6.1 standard deviation score (SDS); 5.7 kg, -3.4 SDS). Assessment, however, revealed GHIS with absent GH-binding protein. Thereafter, the patient received intermittent recombinant human IGF-I (rhIGF-I; 80 microg/kg twice daily) treatment prepubertally for 5.5 years. Genomic DNA was extracted for genetic analysis and each exon was PCR amplified individually. Further, in order to amplify the GHR gene from exon 4 to 6, Expand Long Template PCR (Roche) was carried out. In addition, RNA isolation and RT-PCR were performed. RESULTS: Separate PCRs of each of the exons of the GHR gene revealed that exon 5 in the patient was missing. Thereafter, "Long PCR" from exons 4 to 6 revealed a 4097 bp deletion encompassing exon 5, in a homozygous state in the patient and in a heterozygous state in both parents. RT-PCR analysis revealed an exact absence of exon 5 resulting in a frameshift, leading to a stop codon in exon 6, which predicts a truncated, non-functional GHR protein. CONCLUSION: Fifty-one different mutations within the GHR gene causing GHIS have been reported so far. In contrast, only three deletions within the GHR gene are known. We describe a patient suffering from GHIS caused by a novel 4 kb deletion of the GHR gene encompassing exon 5 and, additionally, we focus on the effect of intermittent rhIGF-I treatment during prepuberty. PMID- 15132719 TI - A meiotic recombination in a new isolated familial somatotropinoma kindred. AB - We report here the genetic findings of a new isolated familial somatotropinoma (IFS) kindred in which the mother (subject I:2) and one daughter (subject II:2) are affected; their ages at diagnosis were 25 and 14 years respectively. Additionally, patient I:2 developed virilization due to an androgen-secreting adrenocortical mass, presenting clinical and molecular features of sporadic adrenal carcinoma. To genotype this family and to narrow down the candidate interval of the putative IFS gene at 11q13, we performed haplotyping on the DNA from all five members of the family and allelotyping of one available somatotropinoma using polymorphic microsatellite markers from chromosome region 11q12.1-11q13.5. Results indicated that the disease haplotype, between markers D11S956 and D11S527, was transmitted from subject I:2 only to subject II:2. A meiotic recombination event was detected in the fraternal twin sister of II:2 (subject II:1), but her disease status is unknown. Since she is only 18 years old this genetic event cannot yet narrow down the area involved in the pathogenesis of IFS. Allelotyping of the somatotropinoma from II:2 revealed loss of the chromosome carrying the wild-type copy of the putative IFS gene inherited from her father. These results support the involvement of a tumor suppressor gene at 11q13.1-q13.3 in the pathogenesis of IFS. PMID- 15132720 TI - Diagnostic 131I whole body scanning after thyroidectomy and ablation for differentiated thyroid cancer. AB - OBJECTIVE: To assess the value of the diagnostic whole body (131)I scan after thyroidectomy and (131)I ablation. DESIGN: Retrospective analysis of all patients with differentiated thyroid cancer treated in one centre between 1990 and 2000. RESULTS: A total of 153 consecutive patients who underwent diagnostic scanning following ablative therapy were identified. This diagnostic scan was positive in 20 patients (13%) and faintly positive in 16 patients (11%). The majority (117 patients) had negative scans. Of the 20 patients with positive scans, four received no further treatment, nine showed no abnormal uptake following a second ablative (131)I dose and seven had uptake in the thyroid bed (six) or in neck nodes (one) after repeat ablation. OUTCOME: In the group with positive scans, the four patients who received no further treatment and the nine with a negative second ablation scan remained disease free during follow-up. No patient with a positive diagnostic scan received additional (131)I therapy which would not otherwise have been given based on the clinical findings, serum thyroglobulin (Tg) values or the presence of anti-Tg antibodies. Ten of the patients with negative scans developed recurrent disease which was always detected clinically or by a rising serum Tg value. CONCLUSIONS: Diagnostic whole body (131)I scans add little extra information and in our experience do not influence patient management. They should be reserved for patients in whom serum Tg levels are unreliable because of the presence of antibodies or when there is clinical suspicion of tumour. PMID- 15132721 TI - Effects of TSH-suppressive therapy on cardiac morphology and function: beneficial effects of the addition of beta-blockade on diastolic dysfunction. AB - OBJECTIVE: To investigate the effects of long-term levothyroxine (LT4) suppressive therapy on the heart and the effects of beta-blockade on cardiac functions. DESIGN: Twelve female patients receiving LT4, selected from a group of patients with TSH levels of 0.1-0.4 microU/ml, were evaluated. The control group consisted of 11 healthy subjects and 12 patients with TSH levels <0.05 microU/ml. METHODS: Cardiac evaluation consisted of a 12-lead electrocardiogram and an echocardiographic study. Left ventricular mass index (LVMI), isovolumetric relaxation time (IVR), left ventricular end systolic (LVESD) and diastolic diameters, early (VE) and late (VA) diastolic flow velocities and fractional shortening were evaluated. Exercise capacity was assessed with a bicycle ergometer. Both work load and maximal exercise time were measured. Atenolol was given to the patient group at a dosage of 50 mg/day for 3 months and evaluations were repeated. RESULTS: On basal evaluations, LVMI (96+/-17 vs 78+/-21 g/m(2)) and IVR (101+/-9 vs 91+/-4 ms) were found to be increased in the patients taking LT4 (P<0.01). LVESD was also lower than controls (P<0.05). A decrease in VE and an increase in VA were also observed in the patients (P<0.01). IVR decreased after atenolol (92+/-10 vs 101+/-9 ms, P<0.05). LVMI decreased and VE and VA improved but did not reach statistical significance after beta-blockade. Baseline work load and maximal exercise duration were significantly lower in the patients and improvements were observed after atenolol treatment. CONCLUSIONS: These results indicated that cardiac dysfunction may occur even when TSH is suppressed to 0.1-0.4 microU/ml with LT4. beta-blockade improved the cardiac functions. PMID- 15132723 TI - Dose-, IGF-I- and sex-dependent changes in lipid profile and body composition during GH replacement therapy in adult onset GH deficiency. AB - OBJECTIVE: Patients with GH deficiency of adult onset (GHDA) exhibit dyslipidaemia and increased cardiovascular morbidity. GH replacement potently reduces body fat and serum lipids in GHDA. In recent years, lower GH doses have been introduced. The purpose of this analysis was to explore the response relationship between GH doses, lipids and body composition. DESIGN: Two consecutive, randomized 12-month GH replacement studies covering placebo and three different doses of GH (0.5, 1.0 and 1.7 IU/m(2) per day). Low and intermediate doses were IGF-I titrated. PATIENTS: Fifty-eight patients with severe GHDA, not previously treated with GH and stably substituted for other endocrine deficiencies, were included in the study. METHODS: Serum lipoproteins, serum IGF-I and body composition analysis by dual energy X-ray absorptiometry (DXA) were used. RESULTS: Fifty-seven percent of patients exhibited low density lipoprotein (LDL) cholesterol levels above 4.16 mmol/l, corresponding to the American Heart Association threshold of 160 mg/dl. GH treatment resulted in significant decreases in total and LDL cholesterol, with no significant change in high density lipoprotein cholesterol or triglycerides. The low dose induced no significant changes in lipid levels, whereas the medium dose reduced LDL cholesterol and the high dose decreased both LDL and total cholesterol. The effects depended significantly on the GH dose and the level of IGF-I obtained, but not on gender. GH replacement induced dose-dependent reductions in fat mass and sex-dependent increases in lean mass. CONCLUSIONS: GH given for 1 year at a dosage between 0.5 and 1.7 IU/m(2) per day reduced fat mass in a dose-dependent manner, increased lean body mass and lowered total and LDL cholesterol in patients with severe GHDA. Low dose GH treatment with normal IGF-I levels induced smaller changes compared with high dose therapy, and may need a longer treatment time. PMID- 15132722 TI - Circulating adiponectin levels and cardiovascular risk factors in acromegalic patients. AB - OBJECTIVE: Adiponectin (ApN) is an adipocytokine expressed in human adipose cells with anti-atherogenic and anti-inflammatory properties that plays a role in the pathophysiology of insulin resistance, metabolic syndrome and coronary artery disease. The aim of the study was to evaluate ApN secretion in patients with acromegaly, a chronic disease associated with insulin resistance and increased cardiovascular mortality, and to correlate ApN levels with hormonal, metabolic and cardiovascular parameters. DESIGN AND METHODS: The study included 32 patients with active acromegaly (11 male and 21 female, aged 48+/-11 years, duration of disease: 8+/-6 years, GH: 9.2+/-9.8 microg/l, IGF-I: 80+/-33 nmol/l (means+/ s.d.)) and 38 control subjects sex- and body mass index (BMI)-matched. In all subjects, serum ApN, leptin and ghrelin levels, BMI, waist circumference, insulin resistance (assessed by homeostasis model assessment and the quantitative insulin check index), lipid profile and blood pressure values were evaluated. RESULTS: Acromegalic patients and control subjects had similar ApN levels (9.4+/-3.5 vs 9.5+/-4.0 mg/l, NS), while when considering obese subjects acromegalic patients had ApN levels significantly higher than controls (10.2+/-4 vs 7.5+/-3 mg/l, P<0.05). No significant correlation between ApN and GH/IGF-I levels or duration of disease was found. ApN concentrations negatively correlated with BMI, waist circumference, glucose and diastolic blood pressure and positively with high density lipoprotein cholesterol and ghrelin in controls, while all these correlations were lost in acromegalic patients. CONCLUSIONS: We provide evidence that, although metabolic and cardiovascular abnormalities are present in most acromegalic patients, in these subjects ApN levels are not reduced and, contrary to what is found in BMI-matched controls, do not correlate with cardiovascular risk factors. These data support the view that atherosclerosis is not the main determinant of cardiovascular mortality in acromegaly and suggest a permissive action of GH and/or IGF-I excess on ApN secretion. PMID- 15132724 TI - Circumstances of discovery of phaeochromocytoma: a retrospective study of 41 consecutive patients. AB - OBJECTIVE: Phaeochromocytoma is a rare tumour of the chromaffin cells, the diagnosis of which is based on an assay of metanephrines and treatment is surgical excision of the tumour. It is usually discovered due to a rich and varied symptomatology or classic paroxysmal hypertension. The main purpose of this study was to specify the exact circumstances of discovery of the phaeochromocytomas operated on in our university hospital between 1990 and 2002. DESIGN AND METHODS: Forty-one consecutive and complete case reports of patients who had surgery for phaeochromocytoma were analysed retrospectively. This series includes 10 patients with a genetic disorder predisposing to phaeochromocytoma. RESULTS: The association of headaches and palpitations with sweating was found in only 24% of cases (10/41). Blood pressure anomalies led to the discovery of phaeochromocytoma in only 51% of cases (21/41) and 59% (24/41) of all the patients suffered from hypertension. In almost half the cases (20/41), the tumour was discovered by an imaging method (ultrasonography, CT scan or MRI) which had been performed for reasons unrelated to a blood pressure abnormality. CONCLUSIONS: Phaeochromocytoma, the symptoms of which are not very specific and during which hypertension is present in only half the patients, is a disease that remains rare. Its incidence could be increasing because of changes in the method of detection. Indeed, in our study, different imaging techniques led to its incidental discovery in half of the cases. PMID- 15132726 TI - Increase of serum leptin after short-term pulsatile GnRH administration in children with delayed puberty. AB - OBJECTIVE: Leptin is known to play an important role in pubertal development in humans, probably acting as one permissive factor for the onset of puberty. Leptin serum concentrations change during pubertal development and an initial increase before the onset of puberty has been reported. The underlying mechanism for this increase in leptin levels is unknown. We hypothesized that the pulsatile release of GnRH stimulates leptin metabolism. In this study, the effect of short-term pulsatile GnRH administration on leptin levels in children with delayed onset of puberty was investigated. METHODS: Nineteen children (15 males and four females, mean age 15.5 years, range 13.1-20.5 years), who underwent evaluation for delayed sexual maturation, were included in the study. Sixteen subjects received 36 h of pulsatile intravenous GnRH, using an infusion pump that released 5 microg GnRH every 90 min. Serum concentrations of LH, FSH, testosterone, estradiol and leptin were analysed before and up to 36 h after GnRH administration. Eight patients received a single dose GnRH-agonist stimulation test (buserelin acetate test, 10 microg/kg body weight) with a 24-h follow-up (five patients underwent both tests). RESULTS: Mean (+/-s.e.m.) serum leptin increased significantly (P<0.01) after 36 h of pulsatile GnRH administration (7.26+/-1.35 vs 9.75+/-1.76 ng/ml). In contrast, no increase in leptin concentrations was observed after administration of a single dose of buserelin. CONCLUSIONS: These findings suggested that the increase in serum leptin at the onset of puberty is triggered by the pulsatile release of GnRH. PMID- 15132725 TI - Serum dehydroepiandrosterone sulfate concentration as an indicator of adrenocortical suppression during inhaled steroid therapy in adult asthmatic patients. AB - OBJECTIVE: Supraphysiological doses of exogenous glucocorticosteroids cause adrenocortical suppression. Dehydroepiandrosterone sulfate (DHEA-S) is the most abundant adrenal androgen and estrogen precursor. We studied to what extent inhaled glucocorticosteroid therapy for asthma decreases serum DHEA-S concentrations. DESIGN AND METHODS: We measured serum DHEA-S and cortisol concentrations in 101 adult patients with newly detected mild asthma before and after 2 and 12 weeks of treatment with inhaled glucocorticosteroids. The patients were randomized to receive budesonide 200 microg/day (low dose group, n=50) or 800 microg/day (high dose group, n=51) in two parallel groups double-blindly. RESULTS: In the low dose group, serum DHEA-S concentrations decreased from the baseline by a mean of 8 % (95 % confidence interval (CI), 3-13 %, P<0.01) after 2 weeks of therapy, and by 2 % (95 % CI, 9 % decrease to 5 % increase, NS) after 12 weeks. In the high dose group, the respective decreases were 16 % (95 % CI, 10-21 %, P<0.001) and 18 % (95 % CI, 12-24 %, P<0.001). The difference between the treatment groups was significant at both 2 and 12 weeks. During the 12 week treatment period the baseline concentrations of serum cortisol did not decrease in the low dose group, while in the high dose group the decrease was significant at 12 weeks (P<0.01), but not at 2 weeks. The forced expiratory volume in 1 s improved equally well in both groups. CONCLUSIONS: Inhaled budesonide decreased serum DHEA-S concentrations, which may indicate adrenocortical suppression. Reduced adrenal production of androgen and estrogen precursors may increase the risk of osteoporosis especially in postmenopausal women. PMID- 15132727 TI - A common polymorphism in the 5'-untranslated region of the aromatase gene influences bone mass and fracture risk. AB - OBJECTIVE: The aromatization of androgenic precursors in peripheral tissues, including bone, is the main source of estrogens after the menopause. CYP19, the gene encoding aromatase, has a long 5'-untranslated region with several variants of exon I and specific promoters. The aim of this study was to investigate the possible relationship between a common biallelic (C/G) polymorphism located on exon I.2 and bone mineral density (BMD). DESIGN: This was designed to be an association study between CYP19 polymorphism and BMD and the risk of vertebral fractures in women. METHODS: DNA was extracted from the peripheral blood of 299 women (116 premenopausal and 183 postmenopausal). CYP19 alleles were identified by a method based on the exonuclease activity of Taq-polymerase. BMD was determined by dual-energy absorptiometry. RESULTS: In premenopausal women there were no genotype-related differences in BMD. However, postmenopausal women with the CC genotype had lower spine and hip BMD than those with the GG genotype. The association between CYP19 genotypes and BMD was independent of other variables, such as age, height, body weight, calcium intake or years since menopause. The CC genotype was also associated with an increased risk of osteoporotic vertebral fractures (odds ratio 2.0; P=0.03). Serum levels of estrone and estradiol were similar in women with CC and GG alleles. CONCLUSIONS: A common biallelic polymorphism in the 5'-untranslated region of the CYP19-aromatase gene was associated with significant differences in bone mass and the risk of vertebral fractures in postmenopausal women. Given the frequency of allelic variants, genotype-related differences appear to be important from the perspective of the individual as well as the general population. Further studies are needed to elucidate underlying mechanisms that may be dependent on differences in estrogen bioactivity at the bone tissue level. PMID- 15132728 TI - Effects of tibolone and cyclic hormone replacement therapy on glucose metabolism in non-diabetic obese postmenopausal women: a randomized study. AB - OBJECTIVE AND METHODS: To study the effects of hormone replacement therapy on glucose metabolism, 31 obese (body mass index > or =27 kg/m(2)) postmenopausal women were randomized to treatment with tibolone (2.5 mg once daily; TIB; n=16) or to oestradiol valerate (2 mg daily)-dydrogesterone (20 mg daily for 2 weeks every 3 months; ED; n=15) for 12 months. Oral (OGTTs) and intravenous glucose tolerance tests (IVGTTs) and a euglycaemic hyperinsulinaemic clamp were performed before and at 6 and 12 months of treatment. RESULTS: TIB decreased the rates of whole body glucose uptake (WBGU) at 6 (P=0.04) and 12 months (P<0.001), but it did not have a significant effect on glucose tolerance. In OGTTs, serum insulin and C-peptide concentrations 2 h after the oral glucose load were increased (P<0.001 and P=0.05 respectively) at 12 months of treatment with TIB, but no changes in the areas under the curve (AUC) of insulin or C-peptide were observed. Furthermore, TIB did not have a significant effect on insulin secretion, the metabolic clearance rate (MCR) of insulin or hepatic insulin extraction. Treatment with ED did not modify the rates of WBGU, but it increased the MCR of insulin (P=0.017) and hepatic insulin extraction (P<0.001) and tended to decrease the insulin AUC (P=0.07). Moreover, glucose tolerance slightly deteriorated during this treatment (P=0.02). Although early phase insulin secretion evaluated by the serum C-peptide response at 30 min in the OGTT increased (P=0.046), the first-phase insulin response during the IVGTT decreased (P=0.05) during ED treatment. CONCLUSIONS: Despite the impairment in peripheral insulin sensitivity, TIB treatment had a neutral effect on glucose tolerance, possibly due to a compensatory decrease in endogenous glucose production. The increased demand on insulin induced by ED, due to both a stimulatory effect on pancreatic beta cells and increased insulin metabolism, may explain the slightly detrimental effect on glucose tolerance with this treatment. PMID- 15132729 TI - The associations between plasma adiponectin, ghrelin levels and cardiovascular risk factors. AB - OBJECTIVE: Ghrelin is a recently discovered peptide, which is produced primarily in the stomach. This orexigenic peptide participates not only in the induction of mealtime hunger but also in long-term body weight regulation and energy homeostasis. Adiponectin is a protein secreted by adipocytes, and has been proposed to mediate obesity-related insulin resistance. Moreover, concentrations of adiponectin are reduced in individuals with obesity, insulin resistance and cardiovascular disease. However, human data are sparse about the direct relationship between adiponectin, ghrelin and cardiovascular risk factors including insulin resistance. DESIGN: Three hundred and thirty-eight elderly Korean women (mean age+/-s.d., 72.3+/-5.5 years) were included in the present study. METHODS: Plasma ghrelin and adiponectin levels were measured by RIA. Anthropometric measurements were taken and a 75 g oral glucose tolerance test performed. Fasting insulin and lipid profile were measured and insulin resistance was determined using the homeostasis model assessment insulin resistance index (HOMA-R) and the quantitative insulin sensitivity check index. RESULTS: Plasma adiponectin levels were negatively correlated with central obesity indices such as waist circumference (r=-0.27, P<0.001) and waist-to-hip ratio (WHR) (r=-0.32, P<0.001), and with insulin resistance indices such as fasting insulin (r=-0.17, P=0.004) and HOMA-R (r=-0.13, P=0.035). Plasma ghrelin levels were negatively correlated with WHR (r=-0.12, P=0.03), but plasma adiponectin and ghrelin levels were not correlated (r=0.03, P=0.66). Multiple regression analysis showed that adiponectin was associated with WHR, fasting insulin and fasting glucose levels. When ghrelin was used as a dependent variable, only WHR remained in the final fitted model. CONCLUSION: Fasting plasma adiponectin and ghrelin levels were found to be associated with central obesity or insulin resistance. However, plasma adiponectin and ghrelin concentrations were not associated with each other in elderly Korean women. PMID- 15132730 TI - A randomized trial of nicotinamide and vitamin E in children with recent onset type 1 diabetes (IMDIAB IX). AB - OBJECTIVE: Various adjuvant therapies have been introduced along with intensive insulin therapy in patients with recent onset type 1 diabetes. Nicotinamide (NA), administered at diagnosis of the disease, can have beneficial effects on the clinical remission rate, improve metabolic control and preserve or slightly increase beta-cell function, probably by reducing toxicity due to free oxygen radicals. Vitamin E, a known antioxidant, inhibits lipid peroxidation; this can lead to protection of islet beta cells from the combined effects of interleukin 1, tumor necrosis factor and gamma interferon. The aim of the present study was to investigate whether the addition of vitamin E to NA could improve metabolic control and the residual beta-cell function, as measured by C-peptide secretion, in children and adolescents with recent onset type 1 diabetes; patients were followed-up for 2 years after diagnosis. PATIENTS AND STUDY DESIGN: Recent onset type 1 diabetes patients (n=64, mean age 8.8 years) were recruited by participating centres of the IMDIAB group. Thirty-two patients were randomized to NA (25 mg/kg body weight) plus vitamin E (15 mg/kg body weight); 32 patients acted as controls and received NA only at the same dose as above. Intensive insulin therapy was applied to both treatment groups. RESULTS: There were three drop outs during the 2-year follow-up period. Overall, patients assigned to the NA+vitamin E group or the NA group did not significantly differ in terms of glycated hemoglobin (HbA1c) levels, insulin requirement or baseline C-peptide secretion. Patients diagnosed at an age of less than 9 years showed significantly reduced C-peptide levels compared with those aged over 9 years at diagnosis and at the 2-year follow-up but there were no differences between the NA and NA+vitamin E treated groups. However at 6 months, patients over 9 years of age treated with NA+vitamin E showed significantly higher C-peptide compared with the NA group (P<0.003). In both age groups and in the different treatment groups, C peptide levels found at diagnosis were preserved 2 years later. CONCLUSIONS: The use of NA alone, or in combination with vitamin E, along with intensive insulin therapy is able to preserve baseline C-peptide secretion for up to 2 years after diagnosis. This finding is of particular interest for pre-pubertal children with type 1 diabetes and has never been reported before. PMID- 15132731 TI - Association of leptin receptor polymorphism with insulin resistance. AB - OBJECTIVE: Although the role of the leptin receptor (LEPR) in obesity is well recognized, its role in insulin sensitivity remains unknown. DESIGN: This was a cross-sectional study to investigate the impact of the Q223R polymorphism of the LEPR gene on insulin sensitivity. METHODS: Sixty-seven glucose-tolerant Caucasians were enrolled. The insulin sensitivity index and glucose clearance were measured using a hyperglycemic clamp. Genomic DNA was extracted for genotyping. RESULTS: We identified 20 QQ, 31 QR, and 16 RR subjects. The three genotypic groups were similar in age, gender distribution, body mass index, waist hip ratio, and blood pressure, but differed in the insulin sensitivity index (P=0.0180) and in glucose clearance (P=0.0220). Multivariate analyses showed that the Q223R polymorphism was independently associated with the insulin sensitivity index (P=0.0317) and glucose clearance (P=0.0436). A trend of increasing low density lipoprotein cholesterol concentration was also observed (P=0.0447). CONCLUSION: We have reported the first association of the Q223R polymorphism of the LEPR gene with insulin resistance. PMID- 15132732 TI - Orexin A suppresses in vivo GH secretion. AB - BACKGROUND/AIMS: Orexins (OXs) are a newly described family of hypothalamic neuropeptides. Based on the distribution of OX neurons and their receptors in the brain, it has been postulated that they could play a role in the regulation of neuroendocrine function. GH secretion is markedly influenced by nutritional status and body weight. To investigate the role OX-A plays in the neuroregulation of GH secretion we have studied its effect on spontaneous GH secretion as well as GH responses to GHRH and ghrelin in freely moving rats. Finally, we also assessed the effect of OX-A on in vitro GH secretion. METHODS: We administered OX-A (10 microg, i.c.v.) or vehicle (10 microl, i.c.v.) to freely moving rats. Spontaneous GH secretion was assessed over 6 h with blood samples taken every 15 min. RESULTS: Administration of OX-A led to a decrease in spontaneous GH secretion in comparison with vehicle-treated rats, as assessed by mean GH levels (means+/ s.e.m. 4.2+/-1.7 ng/ml vs 9.4+/-2.2 ng/ml; P<0.05), mean GH amplitude (3.6+/-0.5 ng/ml vs 20.8+/-5.6 ng/ml; P<0.01) and area under the curve (848+/-379 ng/ml per 4 h vs 1957+/-458 ng/ml per 4 h; P<0.05). In contrast, OX-A failed to modify in vivo GH responses to GHRH (10 microg/kg, i.v.) although it markedly blunted GH responses to ghrelin (40 microg/kg, i.v.) (mean peak GH levels: 331+/-71 ng/ml, vehicle, vs 43+/-11 ng/ml in OX-A-treated rats; P<0.01). Finally, OX-A infusion (10(-7), 10(-8) or 10(-9) M) failed to modify in vitro basal GH secretion or GH responses to GHRH, ghrelin and KCl. CONCLUSIONS: These data indicate that OX-A plays an inhibitory role in GH secretion and may act as a bridge among the regulatory signals that are involved in the control of growth, nutritional status and sleep regulation. PMID- 15132733 TI - Orexin A and B levels in the hypothalamus of female rats: the effects of the estrous cycle and age. AB - OBJECTIVE: Orexins have been implicated in the regulation of several physiological functions including reproduction, energy balance and vigilance state. For successful reproduction, the precisely timed hormonal secretions of the estrous cycle must be combined with appropriate nutritional and vigilance states. The steroid- and nutritional state-dependent modulation of LH release by orexins, as well as an increase of vigilance, suggest that orexins may co ordinate these functions in the course of the estrous cycle. DESIGN: We studied the brain tissue levels of orexins in the course of the estrous cycle in young and middle-aged rats. Young cycling rats (3 months old) and irregularly/non cycling (7-9 months old) female rats were inspected for vaginal smears and serum hormone levels. METHODS: Tissue concentrations of orexin A and B were measured in the hypothalamus and lateral hypothalamus on different days of the estrous cycle. RESULTS: Orexin A concentration in the hypothalamus of young cycling rats was higher on the day of proestrus 5-6 h after the lights were switched on than on the other days of the estrous cycle at the same circadian time. Orexin B concentration was higher on both the day of proestrus and the day of estrus as compared with the days of diestrus. The hypothalamic concentrations of both orexin A and B in the non-cycling middle-aged rats were lower than those in cycling rats on the days of proestrus and estrus. CONCLUSIONS: We have concluded that the high hypothalamic concentration of orexins on the day of proestrus may contribute to the LH and prolactin surges. High orexin A levels may also contribute to the decreased amount of sleep on the day of proestrus. PMID- 15132734 TI - Kinetics of intraoperative fluorescence diagnosis of parathyroid glands. AB - OBJECTIVE: Identification of parathyroid glands is often a challenge even for experienced surgeons. The feasibility and efficacy of fluorescence diagnosis for localization of parathyroids has already been proven in an experimental setting. In preparation for a clinical application of this technique in patients undergoing surgery for hyperparathyroidism, we evaluated the kinetics of fluorescence diagnosis. DESIGN AND METHODS: Fifty rats were randomized into eight groups with different photosensitization parameters using the photosensitizer aminolevulinic acid (ALA): a peritoneal lavage was performed with either 1.5 or 3.0% ALA solution and the induced photosensitization times varied from 0.5, 1 and 2 to 4 h. Under special fluorescence illumination, D-light, the exposed operative site with thyroid, parathyroid glands and neck muscles was examined. The identified parathyroid glands were studied according to fluorescence intensity by spectrometric measurement and compared with surrounding tissue. RESULTS: Photosensitizer accumulation in parathyroid glands, indirectly measured by spectrometry, was up to 3.2 times higher than in thyroid and 2.6 times higher than in muscle tissue (2 h photosensitization with 3.0% ALA). Using 1.5% ALA, the optimum fluorescence intensity and ratio/contrast was slightly lower (parathyroid to-thyroid ratio x 3.0, parathyroid-to-muscle ratio x 1.9) but was reached earlier (1 h) and hence considered as the parameter of choice for a clinical application. CONCLUSIONS: In future clinical application, intraoperative fluorescence diagnosis is expected to increase the ease of identification of atypically located or supernumeric glands. In combination with preoperative diagnostics, this may result in reduced operation time and avoidance of persistent hypercalcaemia. PMID- 15132735 TI - n-3 Fatty acids and cardiovascular disease: evidence explained and mechanisms explored. AB - Long chain n-3 PUFAs (polyunsaturated fatty acids) are found in fatty fish and in fish oils. Substantial evidence from epidemiological and case-control studies indicates that consumption of fish, fatty fish and long-chain n-3 PUFAs reduces the risk of cardiovascular mortality. Secondary prevention studies using long chain n-3 PUFAs in patients post-myocardial infarction have shown a reduction in total and cardiovascular mortality, with an especially potent effect on sudden death. Long-chain n-3 PUFAs have been shown to decrease blood triacylglycerol (triglyceride) concentrations, to decrease production of chemoattractants, growth factors, adhesion molecules, inflammatory eicosanoids and inflammatory cytokines, to lower blood pressure, to increase nitric oxide production, endothelial relaxation and vascular compliance, to decrease thrombosis and cardiac arrhythmias and to increase heart rate variability. These mechanisms most likely explain the primary and secondary cardiovascular protection afforded by long chain n-3 PUFA consumption. A recent study suggests that long-chain n-3 PUFAs might also act to stabilize advanced atherosclerotic plaques, perhaps through their anti-inflammatory effects. As a result of the robust evidence in their favour, a number of recommendations to increase intake of long-chain n-3 PUFAs have been made. PMID- 15132736 TI - Characterization of Prismalin-14, a novel matrix protein from the prismatic layer of the Japanese pearl oyster (Pinctada fucata). AB - The mollusc shell is a hard tissue consisting of calcium carbonate and organic matrices. The organic matrices are believed to play important roles in shell formation. In the present study, we extracted and purified a novel matrix protein, named Prismalin-14, from the acid-insoluble fraction of the prismatic layer of the shell of the Japanese pearl oyster (Pinctada fucata), and determined its whole amino acid sequence by a combination of amino acid sequence analysis and MS analysis of the intact protein and its enzymic digests. Prismalin-14 consisted of 105 amino acid residues, including PIYR repeats, a Gly/Tyr-rich region and N- and C-terminal Asp-rich regions. Prismalin-14 showed inhibitory activity on calcium carbonate precipitation and calcium-binding activity in vitro. The scanning electron microscopy images revealed that Prismalin-14 affected the crystallization of calcium carbonate in vitro. A cDNA encoding Prismalin-14 was cloned and its expression was analysed. The amino acid sequence deduced from the nucleotide sequence of Prismalin-14 cDNA was identical with that determined by peptide sequencing. Northern-blot analysis showed that a Prismalin 14 mRNA was expressed only at the mantle edge. In situ hybridization demonstrated that a Prismalin-14 mRNA was expressed strongly in the inner side of the outer fold of the mantle. These results suggest that Prismalin-14 is a framework protein that plays an important role in the regulation of calcification of the prismatic layer of the shell. PMID- 15132737 TI - ERK5 is targeted to myocyte enhancer factor 2A (MEF2A) through a MAPK docking motif. AB - One critical component in determining the specificity, and efficiency of MAPK (mitogen-activated protein kinase) substrate phophorylation is the presence of distinct docking domains in the substrate proteins. Docking domains have been shown to be important for the activities of members of the ERK (extracellular signal-regulated kinase), JNK (c-Jun N-terminal kinase) and p38 subfamilies of MAPKs towards their substrates. Here, we demonstrate that docking domains also play an important role in ERK5-mediated substrate phosphorylation. The presence of a docking domain promotes both phosphorylation of myocyte enhancer factor, MEF2A, in vitro and its activation in vivo by ERK5. Mutational analysis of the MEF2A docking domain demonstrates that the specificity determinants for ERK5 are similar to those observed with members of the p38 subfamily. A docking domain recognized by ERK5 can direct ERK5 to activate heterologous substrates. Deletion analysis demonstrates that as with other MAPKs, it is the catalytic domain of ERK5 that recognizes the docking domain. Our data therefore extend previous observations on other MAPKs and demonstrate that the requirement for specific docking domains in promoting MAPK action towards substrates is a general property of MAPKs. PMID- 15132738 TI - Hitching a lift hydrodynamically--in swimming, flying and cycling. AB - Swimming animals set the water around them moving, and flying animals generate air movements. Other animals traveling with them can save energy by exploiting these movements of the fluid medium; similarly, a cyclist can save energy by riding close behind another. A new study of dolphin mothers and calves exemplifies the advantages of moving in concert. PMID- 15132739 TI - Examining dolphin hydrodynamics provides clues to calf-loss during tuna fishing. PMID- 15132740 TI - The hydrodynamics of dolphin drafting. AB - BACKGROUND: Drafting in cetaceans is defined as the transfer of forces between individuals without actual physical contact between them. This behavior has long been surmised to explain how young dolphin calves keep up with their rapidly moving mothers. It has recently been observed that a significant number of calves become permanently separated from their mothers during chases by tuna vessels. A study of the hydrodynamics of drafting, initiated in the hope of understanding the mechanisms causing the separation of mothers and calves during fishing related activities, is reported here. RESULTS: Quantitative results are shown for the forces and moments around a pair of unequally sized dolphin-like slender bodies. These include two major effects. First, the so-called Bernoulli suction, which stems from the fact that the local pressure drops in areas of high speed, results in an attractive force between mother and calf. Second is the displacement effect, in which the motion of the mother causes the water in front to move forwards and radially outwards, and water behind the body to move forwards to replace the animal's mass. Thus, the calf can gain a 'free ride' in the forward-moving areas. Utilizing these effects, the neonate can gain up to 90% of the thrust needed to move alongside the mother at speeds of up to 2.4 m/sec. A comparison with observations of eastern spinner dolphins (Stenella longirostris) is presented, showing savings of up to 60% in the thrust that calves require if they are to keep up with their mothers. CONCLUSIONS: A theoretical analysis, backed by observations of free-swimming dolphin schools, indicates that hydrodynamic interactions with mothers play an important role in enabling dolphin calves to keep up with rapidly moving adult school members. PMID- 15132741 TI - Evaluation of developmental phenotypes produced by morpholino antisense targeting of a sea urchin Runx gene. AB - BACKGROUND: Runx transcription factors are important regulators of metazoan development. The sea urchin Runx gene SpRunt was previously identified as a trans activator of the CyIIIa actin gene, a differentiation marker of larval aboral ectoderm. Here we extend the functional analysis of SpRunt, using morpholino antisense oligonucleotides (morpholinos) to interfere with SpRunt expression in the embryo. RESULTS: The developmental effects of four different SpRunt-specific morpholinos were evaluated. The two morpholinos most effective at knocking down SpRunt produce an identical mitotic catastrophe phenotype at late cleavage stage that is an artifact of coincidental mis-targeting to histone mRNA, providing a cautionary example of the insufficiency of two different morpholinos as a control for specificity. The other two morpholinos produce gastrula stage proliferation and differentiation defects that are rescued by exogenous SpRunt mRNA. The expression of 22 genes involved in cell proliferation and differentiation was analyzed in the latter embryos by quantitative polymerase chain reaction. Knockdown of SpRunt was found to perturb the expression of differentiation markers in all of the major tissue territories as well as the expression of cell cycle control genes, including cyclin B and cyclin D. CONCLUSIONS: SpRunt is essential for embryonic development, and is required globally to coordinate cell proliferation and differentiation. PMID- 15132742 TI - A conserved lysine in the estrogen receptor DNA binding domain regulates ligand activation profiles at AP-1 sites, possibly by controlling interactions with a modulating repressor. AB - BACKGROUND: Estrogen receptors alpha and beta (ERalpha and ERbeta) differentially activate genes with AP-1 elements. ERalpha activates AP-1 targets via activation functions with estrogens (the AF-dependent pathway), whereas ERbeta, and a short version of ERalpha (ERalpha DBD-LBD) activate only with anti-estrogens (AF independent pathway). The DNA binding domain (DBD) plays an important role in both pathways, even though neither pathway requires ERE recognition. RESULTS: Mutations of a highly conserved DBD lysine (ERalpha.K206A/G), lead to super activation of AP-1 through activation function dependent pathways, up to 200 fold. This super-activity can be elicited either through ER AFs 1 or 2, or that of a heterologous activation function (VP16). The homologous substitution in ERbeta, K170A, or in ERalpha DBD-LBD leads to estrogen-dependent AP-1 activation and loss of the usually potent anti-estrogen effects. Each of numerous K206 substitutions in ERalpha, except K206R, eliminates anti-estrogen activation and this loss correlates perfectly with a loss of ability to titrate a repressive function from the RU486 bound progesterone receptor. CONCLUSION: We conclude that ER DBDs contain a complex regulatory function that influences ligand activation profiles at AP-1. This function, which requires the integrity of the conserved lysine, both allows for activation at AP-1 with anti-estrogens (with ERbeta and ERalpha DBD-LBD), and prevents ERalpha from becoming superactive at AP-1 with estrogens. We discuss the possibility that a repressor interaction with the DBD both mediates the AF-independent pathway and dampens the AF dependent pathway. Mutations in the conserved lysine might, by this model, disrupt the binding or function of the repressor. PMID- 15132743 TI - Prostate stem cell antigen (PSCA) expression in human prostate cancer tissues and its potential role in prostate carcinogenesis and progression of prostate cancer. AB - BACKGROUND: Prostate stem cell antigen (PSCA) is a recently defined homologue of the Thy-1/Ly-6 family of glycosylphosphatidylinositol (GPI)-anchored cell surface antigens. The purpose of the present study was to examine the expression status of PSCA protein and mRNA in clinical specimens of human prostate cancer (Pca) and to validate it as a potential molecular target for diagnosis and treatment of Pca. MATERIALS AND METHODS: Immunohistochemical (IHC) and in situ hybridization (ISH) analyses of PSCA expression were simultaneously performed on paraffin embedded sections from 20 benign prostatic hyperplasia (BPH), 20 prostatic intraepithelial neoplasm (PIN) and 48 prostate cancer (Pca) tissues, including 9 androgen-independent prostate cancers. The level of PSCA expression was semiquantitatively scored by assessing both the percentage and intensity of PSCA positive staining cells in the specimens. Then compared PSCA expression between BPH, PIN and Pca tissues and analysed the correlations of PSCA expression level with pathological grade, clinical stage and progression to androgen-independence in Pca. RESULTS: In BPH and low grade PIN, PSCA protein and mRNA staining were weak or negative and less intense and uniform than that seen in HGPIN and Pca. There were moderate to strong PSCA protein and mRNA expression in 8 of 11 (72.7%) HGPIN and in 40 of 48 (83.4%) Pca specimens examined by IHC and ISH analyses, with statistical significance compared with BPH (20%) and low grade PIN (22.2%) samples (p < 0.05, respectively). The expression level of PSCA increased with high Gleason grade, advanced stage and progression to androgen-independence (p < 0.05, respectively). In addition, IHC and ISH staining showed a high degree of correlation between PSCA protein and mRNA overexpression. CONCLUSIONS: Our data demonstrate that PSCA as a new cell surface marker is overexpressed by a majority of human Pca. PSCA expression correlates positively with adverse tumor characteristics, such as increasing pathological grade (poor cell differentiation), worsening clinical stage and androgen-independence, and speculatively with prostate carcinogenesis. PSCA protein overexpression results from upregulated transcription of PSCA mRNA. PSCA may have prognostic utility and may be a promising molecular target for diagnosis and treatment of Pca. PMID- 15132745 TI - Measuring mental health of the Dutch population: a comparison of the GHQ-12 and the MHI-5. AB - BACKGROUND: The objective is to compare the performance of the MHI-5 and GHQ-12, both measures of general mental health. Therefore, we studied the relationship of the GHQ-12 and MHI-5 with sociodemographic characteristics, self-reported visits to general practice and mental health care, and with diagnoses made by the general practitioner. METHODS: Data were used from the Second Dutch National Survey of General Practice, which was carried out in 104 practices. This study combines data from a representative sample of the Dutch population with data from general practice. RESULTS: The agreement between the GHQ-12 and MHI-5 is only moderate. Both instruments are however similarly associated with demographic characteristics (except age), self-reported health care use, and psychological and social diagnoses in general practice. CONCLUSIONS: The performance of the MHI 5 and GHQ-12 in terms of predicting mental health problems and related help seeking behaviour is similar. An advantage of the MHI-5 is that it has been widely used, not only in surveys of mental health, but also in surveys of general health and quality of life, and it is shorter. A disadvantage of the MHI-5 is that there is no cut-off point. We recommend a study to establish a valid, internationally comparable cut-off point. PMID- 15132747 TI - Physically synthesized Ni-Cu nanoparticles for magnetic hyperthermia. AB - BACKGROUND: In this paper, a physical method to prepare copper-nickel alloy particles in the sub-micron range for possible self controlled magnetic hyperthermia treatment of cancer is described. It is reported that an increase in tumor temperature decreases the tumor resistance to chemo- and radiation therapies. Self controlled heating at the tumor site to avoid spot heating is managed by controlling the Curie temperature of the magnetic particles. The process described in this paper to produce the nanomagnetic particles allows for a large scale production of these particles. METHODS: The process used here is mainly composed of melting of the Cu-Ni mixture and ball milling of the resulted bulk alloy. Both mechanical abrasion and continuous grinding were used to break down the bulk amount into the desired particle size. RESULTS: It was found that the desired alloy is composed of 71% nickel and 29% copper by weight. It was observed that the coarse sand-grinded powder has a Curie temperature of 345 K and the fine ball-milled powder shows a temperature of 319 K - 320 K. CONCLUSION: Self regulating magnetic hyperthermia can be achieved by synthesizing nanomagnetic particles with desired Curie temperature. In this study the desired range of Curie temperatures was obtained by combination of melting and ball milling of nickel-copper alloy. PMID- 15132746 TI - A new instrument for measuring anticoagulation-related quality of life: development and preliminary validation. AB - BACKGROUND: Anticoagulation can reduce quality of life, and different models of anticoagulation management might have different impacts on satisfaction with this component of medical care. Yet, to our knowledge, there are no scales measuring quality of life and satisfaction with anticoagulation that can be generalized across different models of anticoagulation management. We describe the development and preliminary validation of such an instrument - the Duke Anticoagulation Satisfaction Scale (DASS). METHODS: The DASS is a 25-item scale addressing the (a) negative impacts of anticoagulation (limitations, hassles and burdens); and (b) positive impacts of anticoagulation (confidence, reassurance, satisfaction). Each item has 7 possible responses. The DASS was administered to 262 patients currently receiving oral anticoagulation. Scales measuring generic quality of life, satisfaction with medical care, and tendency to provide socially desirable responses were also administered. Statistical analysis included assessment of item variability, internal consistency (Cronbach's alpha), scale structure (factor analysis), and correlations between the DASS and demographic variables, clinical characteristics, and scores on the above scales. A follow-up study of 105 additional patients assessed test-retest reliability. RESULTS: 220 subjects answered all items. Ceiling and floor effects were modest, and 25 of the 27 proposed items grouped into 2 factors (positive impacts, negative impacts, this latter factor being potentially subdivided into limitations versus hassles and burdens). Each factor had a high degree of internal consistency (Cronbach's alpha 0.78-0.91). The limitations and hassles factors consistently correlated with the SF-36 scales measuring generic quality of life, while the positive psychological impact scale correlated with age and time on anticoagulation. The intra-class correlation coefficient for test-retest reliability was 0.80. CONCLUSIONS: The DASS has demonstrated reasonable psychometric properties to date. Further validation is ongoing. To the degree that dissatisfaction with anticoagulation leads to decreased adherence, poorer INR control, and poor clinical outcomes, the DASS has the potential to help identify reasons for dissatisfaction (and positive satisfaction), and thus help to develop interventions to break this cycle. As an instrument designed to be applicable across multiple models of anticoagulation management, the DASS could be crucial in the scientific comparison between those models of care. PMID- 15132744 TI - Barrett's oesophagus and adenocarcinoma. PMID- 15132748 TI - Detection of hepatitis C virus in the nasal secretions of an intranasal drug user. AB - BACKGROUND: One controversial source of infection for hepatitis C virus (HCV) involves the sharing of contaminated implements, such as straws or spoons, used to nasally inhale cocaine and other powdered drugs. An essential precondition for this mode of transmission is the presence of HCV in the nasal secretions of intranasal drug users. METHODS: Blood and nasal secretion samples were collected from five plasma-positive chronic intranasal drug users and tested for HCV RNA using RT-PCR. RESULTS: HCV was detected in all five blood samples and in the nasal secretions of the subject with the highest serum viral load. CONCLUSIONS: This study is the first to demonstrate the presence of HCV in nasal secretions. This finding has implications for potential transmission of HCV through contact with contaminated nasal secretions. PMID- 15132749 TI - Experimental system to displace radioisotopes from upper to deeper soil layers: chemical research. AB - BACKGROUND: Radioisotopes are introduced into the environment following nuclear power plant accidents or nuclear weapons tests. The immobility of these radioactive elements in uppermost soil layers represents a problem for human health, since they can easily be incorporated in the food chain. Preventing their assimilation by plants may be a first step towards the total recovery of contaminated areas. METHODS: The possibility of displacing radionuclides from the most superficial soil layers and their subsequent stabilisation at lower levels were investigated in laboratory trials. An experimental system reproducing the environmental conditions of contaminated areas was designed in plastic columns. A radiopolluted soil sample was treated with solutions containing ions normally used in fertilisation (NO3-, NH4+, PO4--- and K+). RESULTS: Contaminated soils treated with an acid solution of ions NO3-, PO4--- and K+, undergo a reduction of radioactivity up to 35%, after a series of washes which simulate one year's rainfall. The capacity of the deepest soil layers to immobilize the radionuclides percolated from the superficial layers was also confirmed. CONCLUSION: The migration of radionuclides towards deeper soil layers, following chemical treatments, and their subsequent stabilization reduces bioavailability in the uppermost soil horizon, preventing at the same time their transfer into the water bearing stratum. PMID- 15132750 TI - A new method for detection of pfmdr1 mutations in Plasmodium falciparum DNA using real-time PCR. AB - BACKGROUND: Surveillance for drug-resistant Plasmodium falciparum should be a component of malaria control programmes. Real-time PCR methods for the detection of parasite single-nucleotide polymorphisms (SNPs) and gene amplification could be useful survellance tools. METHODS: A real-time PCR assay has been developed that identifies single nucleotide polymorphisms (SNPs) at amino acids 86, 184, 1034 and 1042 in the P. falciparum multi-drug resistant (pfmdr 1) gene that may be associated with anti-malarial drug resistance. RESULTS: This assay has a sensitivity and specificity of 94% and 100% when compared to traditional PCR methods for genotyping. Only 54 of 68 (79%) paired pre- and post-culture DNA samples were concordant at all four loci. CONCLUSION: Real-time PCR is a sensitive and specific method to detect SNP's in pfmdr 1. Genotypes of parasites after in vitro culture may not reflect that seen in vivo. PMID- 15132752 TI - Screening family planning needs: an operations research project in Guatemala. AB - BACKGROUND: Public sector health care providers in rural Guatemala have infrequently offered family planning information and services in routine visits. This operations research project tested a strategy to modify certain practices that prevent health workers from proactively screening clients' needs and meeting them. METHODS: The research design was quasi-experimental with a pretest-posttest follow-up comparison group design. Health districts, which comprise health centers and posts, were purposively assigned to intervention or comparison groups to assure comparability of the two groups. The strategy was based on a job-aid designed to guide health workers in screening clients' reproductive intentions and family planning needs, help them to offer contraceptive methods if the woman expressed interest, and facilitate the provision of the method chosen at the time of the visit. The strategy was implemented at intervention sites during a period of six months. Upon completion of post-intervention measurements, the strategy was scaled up to the comparison sites, and a follow-up assessment was conducted nine months later. Results were evaluated by conducting three rounds of exit interviews with women exposed to the risk of unwanted pregnancy. RESULTS: Study results showed a two to five-fold increase in providers' screening of clients' reproductive intentions. The proportion of clients who received information about contraceptives increased from 8% at the baseline to 42% immediately post intervention, and 36% at the follow-up survey. The intervention also proved successful in improving the role service providers play in offering women a chance to ask questions and assisting women in making a selection. The proportion of women who received a method, referral or appointment increased and remained high in the intervention group, although no change was seen in the comparison group after their participation in the strategy. CONCLUSION: The easy-to-use job aid developed for this project proved useful for screening clients' needs and reducing providers' reluctance to discuss family planning with clients and offer contraceptive services. Such family planning screening devices can be useful in traditional settings where both providers and clients shy away from discussing family planning issues. PMID- 15132751 TI - Protein quality control in the bacterial periplasm. AB - The proper functioning of extracytoplasmic proteins requires their export to, and productive folding in, the correct cellular compartment. All proteins in Escherichia coli are initially synthesized in the cytoplasm, then follow a pathway that depends upon their ultimate cellular destination. Many proteins destined for the periplasm are synthesized as precursors carrying an N-terminal signal sequence that directs them to the general secretion machinery at the inner membrane. After translocation and signal sequence cleavage, the newly exported mature proteins are folded and assembled in the periplasm. Maintaining quality control over these processes depends on chaperones, folding catalysts, and proteases. This article summarizes the general principles which control protein folding in the bacterial periplasm by focusing on the periplasmic maltose-binding protein. PMID- 15132753 TI - Endurance training of respiratory muscles improves cycling performance in fit young cyclists. AB - BACKGROUND: Whether or not isolated endurance training of the respiratory muscles improves whole-body endurance exercise performance is controversial, with some studies reporting enhancements of 50% or more, and others reporting no change. Twenty fit (VO2 max 56.0 ml/kg/min), experienced cyclists were randomly assigned to three groups. The experimental group (n = 10) trained their respiratory muscles via 20, 45 min sessions of hyperpnea. The placebo group (n = 4) underwent "sham" training (20, 5 min sessions), and the control group (n = 6) did no training. RESULTS: After training, the experimental group increased their respiratory muscle endurance capacity by 12%. Performance on a bicycle time trial test designed to last about 40 min improved by 4.7% (9 of 10 subjects showed improvement). There were no test-re-test improvements in either respiratory muscle or bicycle exercise endurance performance in the placebo group, nor in the control group. After training, the experimental group had significantly higher ventilatory output and VO2, and lower PCO2, during constant work-rate exercise; the placebo and control groups did not show these changes. The perceived respiratory effort was unchanged in spite of the higher ventilation rate after training. CONCLUSIONS: The results suggest that respiratory muscle endurance training improves cycling performance in fit, experienced cyclists. The relative hyperventilation with no change in respiratory effort sensations suggest that respiratory muscle training allows subjects to tolerate the higher exercise ventilatory response without more dyspnea. Whether or not this can explain the enhanced performance is unknown. PMID- 15132754 TI - Intrinsic and climatic factors in North-American animal population dynamics. AB - BACKGROUND: Extensive work has been done to identify and explain multi-year cycles in animal populations. Several attempts have been made to relate these to climatic cycles. We use advanced time series analysis methods to attribute cyclicities in several North-American mammal species to abiotic vs. biotic factors. RESULTS: We study eleven century-long time series of fur-counts and three climatic records--the North Atlantic Oscillation (NAO), the El-Nino Southern Oscillation (ENSO), and Northern Hemisphere (NH) temperatures--that extend over the same time interval. Several complementary methods of spectral analysis are applied to these 14 times series, singly or jointly. These spectral analyses were applied to the leading principal components (PCs) of the data sets. The use of both PC analysis and spectral analysis helps distinguish external from intrinsic factors that influence the dynamics of the mammal populations. CONCLUSIONS: Our results show that all three climatic indices influence the animal-population dynamics: they explain a substantial part of the variance in the fur-counts and share characteristic periods with the fur-count data set. In addition to the climate-related periods, the fur-count time series also contain a significant 3-year period that is, in all likelihood, caused by biological interactions. PMID- 15132755 TI - Computerized adaptive measurement of depression: a simulation study. AB - BACKGROUND: Efficient, accurate instruments for measuring depression are increasingly important in clinical practice. We developed a computerized adaptive version of the Beck Depression Inventory (BDI). We examined its efficiency and its usefulness in identifying Major Depressive Episodes (MDE) and in measuring depression severity. METHODS: Subjects were 744 participants in research studies in which each subject completed both the BDI and the SCID. In addition, 285 patients completed the Hamilton Depression Rating Scale. RESULTS: The adaptive BDI had an AUC as an indicator of a SCID diagnosis of MDE of 88%, equivalent to the full BDI. The adaptive BDI asked fewer questions than the full BDI (5.6 versus 21 items). The adaptive latent depression score correlated r =.92 with the BDI total score and the latent depression score correlated more highly with the Hamilton (r =.74) than the BDI total score did (r =.70). CONCLUSIONS: Adaptive testing for depression may provide greatly increased efficiency without loss of accuracy in identifying MDE or in measuring depression severity. PMID- 15132757 TI - The relationship between sensory impairment and functional independence among elderly. AB - BACKGROUND: It has been well established that increasing age is associated with decreasing functional ability in older adults. It is important to understand the specific factors that affect instrumental activities of daily living (IADL) and functional independence among older adults with sensory disabilities. METHODS: Nationally representative sample of adults aged 55 years and older with seeing or hearing disabilities were categorised into three sensory classifications: "Seeing Disabled but Hearing Abled" (SD-HA), "Hearing Disabled but Seeing Abled" (HD-SA), and both "Seeing and Hearing Disabled" (SD-HD). The additional category of "Seeing Disabled and/or Hearing Disabled" (SD and/or HD) was created to calculate the total of all individuals from the above categories who either had a seeing or hearing disability or both sensory disabilities. Respondents were asked to indicate whether they received assistance in performing seven IADL and their level of functional independence. RESULTS: The most common factors that affect IADL were heavy chores, grocery shopping and housework. Individuals with both seeing and hearing disabilities (SD-HD) reported having the most IADL restrictions, followed by individuals with only seeing disabilities (SD-HA) and only hearing disabilities (HD-SA). Individuals with severe sensory disabilities were generally more likely to report IADL restrictions and less likely to have decision-making control and be happy with their lives. In each sensory classification, females aged 55-64 years and 65 years and older reported more IADL restrictions than males. CONCLUSION: Both seeing and hearing disabilities have a significant impact on restricting an individual's IADL. PMID- 15132756 TI - Recruitment of focal adhesion kinase and paxillin to beta1 integrin promotes cancer cell migration via mitogen activated protein kinase activation. AB - BACKGROUND: Integrin-extracellular matrix interactions activate signaling cascades such as mitogen activated protein kinases (MAPK). Integrin binding to extracellular matrix increases tyrosine phosphorylation of focal adhesion kinase (FAK). Inhibition of FAK activity by expression of its carboxyl terminus decreases cell motility, and cells from FAK deficient mice also show reduced migration. Paxillin is a focal adhesion protein which is also phosphorylated on tyrosine. FAK recruitment of paxillin to the cell membrane correlates with Shc phosphorylation and activation of MAPK. Decreased FAK expression inhibits papilloma formation in a mouse skin carcinogenesis model. We previously demonstrated that MAPK activation was required for growth factor induced in vitro migration and invasion by human squamous cell carcinoma (SCC) lines. METHODS: Adapter protein recruitment to integrin subunits was examined by co immunoprecipitation in SCC cells attached to type IV collagen or plastic. Stable clones overexpressing FAK or paxillin were created using the lipofection technique. Modified Boyden chambers were used for invasion assays. RESULTS: In the present study, we showed that FAK and paxillin but not Shc are recruited to the beta1 integrin cytoplasmic domain following attachment of SCC cells to type IV collagen. Overexpression of either FAK or paxillin stimulated cancer cell migration on type IV collagen and invasion through reconstituted basement membrane which was dependent on MAPK activity. CONCLUSIONS: We concluded that recruitment of focal adhesion kinase and paxillin to beta1 integrin promoted cancer cell migration via the mitogen activated protein kinase pathway. PMID- 15132759 TI - Adaptation of a South American malaria vector to laboratory colonization suggests faster-male evolution for mating ability. AB - BACKGROUND: Anopheles (Nyssorhynchus) albitarsis (Diptera: Culicidae) is one of the very few South American mosquito vectors of malaria successfully colonized in the laboratory. These vectors are very hard to breed because they rarely mate in artificial conditions. A few years ago a free-mating laboratory colony of An. albitarsis sensu stricto was established after about 30 generations of artificial mating. To begin to understand the process of adaptation of these malaria vectors to the laboratory we have compared the insemination rates of colony mosquitoes to those from the original population in both artificial and free-mating crosses. We also carried out crossing experiments between the two types of mosquitoes for a preliminary analysis of the genetic basis of such adaptation. RESULTS: We show that, compared to the original population, colony males but not females have increased their insemination rates in the laboratory in both types of mating, suggesting that faster-male evolution of mating ability might have occurred during the colonization process. CONCLUSIONS: The results are consistent with the faster-male theory, which predicts that sexual selection will cause faster rates of evolution of genes expressed in males. The data also suggests that attempts to colonize other South American malaria mosquitoes will be more successful if special attention is given to the male ability to mate in a confined space. PMID- 15132758 TI - Diversity and repertoire of IgW and IgM VH families in the newborn nurse shark. AB - BACKGROUND: Adult cartilaginous fish express three immunoglobulin (Ig) isotypes, IgM, IgNAR and IgW. Newborn nurse sharks, Ginglymostoma cirratum, produce 19S (multimeric) IgM and monomeric/dimeric IgM1gj, a germline-joined, IgM-related VH, and very low amounts of 7S (monomeric) IgM and IgNAR proteins. Newborn IgNAR VH mRNAs are diverse in the complementarity-determining region 3 (CDR3) with non templated nucleotide (N-region) addition, which suggests that, unlike in many other vertebrates, terminal deoxynucleotidyl transferase (TdT) expressed at birth is functional. IgW is present in the lungfish, a bony fish sharing a common ancestor with sharks 460 million years ago, implying that the IgW VH family is as old as the IgM VH family. This nurse shark study examined the IgM and IgW VH repertoire from birth through adult life, and analyzed the phylogenetic relationships of these gene families. RESULTS: IgM and IgW VH cDNA clones isolated from newborn nurse shark primary and secondary lymphoid tissues had highly diverse and unique CDR3 with N-region addition and VDJ gene rearrangement, implicating functional TdT and RAG gene activity. Despite the clear presence of N region additions, newborn CDR3 were significantly shorter than those of adults. The IgM clones are all included in a conventional VH family that can be classified into five discrete groups, none of which is orthologous to IgM VH genes in other elasmobranchs. In addition, a novel divergent VH family was orthologous to a published monotypic VH horn shark family. IgW VH genes have diverged sufficiently to form three families. IgM and IgW VH serine codons using the potential somatic hypermutation hotspot sequence occur mainly in VH framework 1 (FR1) and CDR1. Phylogenetic analysis of cartilaginous fish and lungfish IgM and IgW demonstrated they form two major ancient gene groups; furthermore, these VH genes generally diversify (duplicate and diverge) within a species. CONCLUSION: As in ratfish, sandbar and horn sharks, most nurse shark IgM VH genes are from one family with multiple, heterogeneous loci. Their IgW VH genes have diversified, forming at least three families. The neonatal shark Ig VH CDR3 repertoire, diversified via N-region addition, is shorter than the adult VDJ junction, suggesting one means of postnatal repertoire diversification is expression of longer CDR3 junctions. PMID- 15132760 TI - eL-DASionator: an LDAS upload file generator. AB - BACKGROUND: The Distributed Annotation System (DAS) allows merging of DNA sequence annotations from multiple sources and provides a single annotation view. A straightforward way to establish a DAS annotation server is to use the "Lightweight DAS" server (LDAS). Onto this type of server, annotations can be uploaded as flat text files in a defined format. The popular Ensembl ContigView uses the same format for the transient upload and display of user data. RESULTS: In order to easily generate LDAS upload files we developed a software tool that is accessible via a web-interface http://atgc.lirmm.fr/eldasionator.html. Users can submit their DNA sequences of interest. Our program (i) aligns these sequences to the reference sequences of Ensembl, (ii) determines start and end positions of each sequence on the reference sequence, and (iii) generates a formatted annotation file. This file can be used to load any LDAS annotation server or it can be uploaded to the Ensembl ContigView. CONCLUSION: The eL DASionator is an on-line tool that is intended for life-science researchers with little bioinformatics background. It conveniently generates LDAS upload files, and makes it possible to generate annotations in a standard format that permits comfortable sharing of this data. PMID- 15132761 TI - REM sleep and cortisol responses to scopolamine during depression and remission in women. AB - Baseline electroencephalographic (EEG) sleep and the EEG sleep response to scopolamine were studied in 10 adult female patients with unipolar major depressive disorder. Subjects were studied twice for two consecutive nights while depressed and, again, during remission. On the second night of each two-night session, normal saline or scopolamine (1.5 microg/kg, i.m.) was administered in a randomized, double-blind, cross-over fashion. Nocturnal urinary free cortisol (NUFC) measures also were collected. Compared to the depressed state, NUFC was significantly lower during remission. In contrast, baseline EEG sleep measures did not differ from episode to remission. Scopolamine suppressed rapid eye movement (REM) sleep to a comparable extent during the depressive episode and in remission. Scopolamine also reduced NUFC secretion during both clinical states, but to a lesser extent than REM sleep suppression. The findings suggest that the dysregulation in cholinergic systems associated with depressive illness may be persistent during remission, at least for some cholinergic systems. The results also suggest that the central cholinergic system(s) that regulate(s) REM sleep may be more sensitive to dysregulation than the cholinergic system(s) that control(s) nocturnal cortisol secretion. PMID- 15132762 TI - Increased repetitive behaviours and prolactin responsivity to oral m chlorophenylpiperazine in adults with autism spectrum disorders. AB - Autism is a neurodevelopmental disorder characterized by dysfunction in three primary behavioural domains: repetitive behaviours, social deficits, and language abnormalities. There is evidence that abnormalities exist in the serotonin (5-HT) system in autism spectrum patients. Furthermore, 5-HT is known to play a role in repetitive and social behaviours. This study examined the effect of m chlorophenylpiperazine (m-CPP) on repetitive behaviours and prolactin response in 11 adults with autism or Aspergers disorder and 8 age- and gender-matched healthy controls via randomized double-blind, m-CPP and placebo challenges. The primary outcome measure was an instrument rating six repetitive behaviours: need to know, repeating, ordering, need to tell/ask, self-injury, and touching. Patients with autism spectrum disorders showed a significant increase in repetitive behaviours at end-point following oral m-CPP in comparison to placebo. Additionally subjects with autism spectrum disorders showed a significantly increased prolactin response to m-CPP compared to normal controls, with neither group responding to placebo. This study provides further evidence for altered 5-HT sensitivity in individuals with autism spectrum disorders, as well as a possible relationship between repetitive behaviours in autism spectrum disorders and abnormalities in the 5-HT system. PMID- 15132764 TI - Search for new biomarkers of gastric cancer through serial analysis of gene expression and its clinical implications. AB - Gastric cancer is one of the most common human cancers and is the second most frequent cause of cancer-related death in the world. Serial analysis of gene expression (SAGE) is a powerful technique to allow genome-wide analysis of gene expression in a quantitative manner without prior knowledge of the gene sequences. SAGE on 5 samples of gastric cancer with different histology and clinical stages have created large SAGE libraries of gastric cancer that enable us to identify new cancer biomarkers. Commonly up-regulated genes in gastric cancer in comparison with normal gastric epithelia included CEACAM6, APOC1 and YF13H12. By comparing gene expression profiles of gastric cancers at early and advanced stages, several genes differentially expressed by tumor stage were also identified, including FUS, CDH17, COL1A1 and COL1A2, which should be novel genetic markers for high-grade malignancy. Regenerating gene type IV (REGIV) is one of the most up-regulated genes in a SAGE library of a scirrhous-type gastric cancer. In vitro studies using RegIV-transfected cells revealed that RegIV is secreted by cancer cells and inhibits apoptosis, suggesting that RegIV may serve as a novel biomarker and therapeutic target for gastric cancer. Production of RNA aptamers could be a useful approach to establish a detection system in blood. A custom-made array, named Ex-STOMACHIP, consisting of 395 genes, including highly differentially expressed genes identified by our SAGE and other known genes related to carcinogenesis and chemosensitivity, is useful to study the molecular pathogenesis of gastric cancer and to obtain information about biological behavior and sensitivity to therapy in the clinical setting. Combined analyses of gene expression profile, genetic polymorphism and genetic instability will aid not only cancer detection, but also characterization of individual cancers and patients, leading to personalized medicine and cancer prevention. PMID- 15132763 TI - Carbohydrate-mediated cell adhesion in cancer metastasis and angiogenesis. AB - Malignant transformation is associated with abnormal glycosylation, resulting in the synthesis and expression of altered carbohydrate determinants including sialyl Lewisa and sialyl Lewisx. The sialyl Lewisa and sialyl Lewisx determinants appear in the sera of patients with cancer, and are extensively utilized for serum diagnosis of cancers in Japan. Sialyl Lewisa and sialyl Lewisx are involved in selectin-mediated adhesion of cancer cells to vascular endothelium, and these determinants are thought to be closely associated with hematogenous metastasis of cancers. Recent progress in this area includes the following: 1. Substantial increases in solid clinical statistics that further confirm the contribution of these determinants in the progression of a wide variety of cancers; 2. Elucidation of the ligand specificity of the three family members of selectins and evaluation of the roles of these molecules in cancer cell adhesion; and 3. Advances in the study of the mechanism that leads to the enhanced expression of the sialyl Lewis(a/x) determinants in malignant cells. These recent results have confirmed that these determinants are not merely markers for cancers, but are functionally implicated in the malignant behavior of cancer cells. The results also suggested that the increase of these determinants in malignant cells is an inevitable consequence of the malignant transformation of cells. Considerable new knowledge has also been accumulated regarding the therapeutic implications for suppression of hematogenous metastasis targeting this cell adhesion system. PMID- 15132765 TI - Dose-related changes of oxidative stress and cell proliferation in kidneys of male and female F344 rats exposed to potassium bromate. AB - It is still of importance to investigate renal carcinogenesis by potassium bromate (KBrO3), a by-product of water disinfection by ozonation, for assessment of the risk to man. Five female F344 rats in each group were given KBrO3 at a dose of 300 mg/kg by single i.g. intubation or at a dose of 80 mg/kg by single i.p. injection, and were killed 48 h after the administration for measurements of thiobarbituric acid-reactive substances (TBARS) and 8-oxodeoxyguanosine (8-oxodG) levels in the kidney. Both levels in the treated animals were significantly elevated as compared with the control values. In a second experiment, 5 male and female F344 rats in each group were administered KBrO3 at concentrations of 0, 15, 30, 60, 125, 250 and 500 ppm in the drinking water for 4 weeks. KBrO3 in the drinking water did not elevate TBARS in either sex at any of the doses examined, but 8-oxodG formation in both sexes at 250 ppm and above was significantly higher than in the controls. Additionally, the bromodeoxyuridine-labeling index for proximal convoluted tubules was significantly increased at 30 ppm and above in the males, and at 250 ppm and above in the females. Alpha2u-globulin accumulation in the kidneys of male rats was increased with statistical significance at 125 ppm and above. These findings suggest that DNA oxidation induced by KBrO3 may occur independently of lipid peroxidation and more than 250 ppm KBrO3 in the drinking water can exert a carcinogenic effect by way of oxidative stress. PMID- 15132766 TI - Preferential mammary carcinogenic effects of 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP) in human c-Ha-ras proto-oncogene transgenic rats. AB - In order to clarify the susceptibility of the Hras128 rat harboring copies of the human c-Ha-ras proto-oncogene to 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), Hras128 rats were intragastically treated with 100 mg/kg PhIP 8 times (females) or 80 mg/kg PhIP 10 times (males) over a 9-week period, then sacrificed at weeks 12 and 30. Multiple mammary tumors of adenocarcinoma type were induced in all females, while 83% of treated males developed adenocarcinomas, sarcomas and transitional carcinosarcomas, as evidenced by casein and vimentin immunoreactivity. All tumors examined had mutations in the c-Ha-ras transgene, while the endogenous rat c-Ha-ras gene was intact. Our results indicate that 1) Hras128 rats of both sexes are preferentially susceptible to mammary carcinogenesis with PhIP; 2) activation of the transgene, but not the endogenous c-Ha-ras gene, may be important in this regard; 3) the variety of tumor types evident in male rats indicates that immature mammary gland cells of the terminal end buds may be a target of PhIP; 4) although the transgene is expressed in all organs, susceptibility to PhIP is limited to mammary glands. PMID- 15132767 TI - Possible enhancing effects of atrazine and nonylphenol on 7,12 dimethylbenz[a]anthracene-induced mammary tumor development in human c-Ha-ras proto-oncogene transgenic rats. AB - Our transgenic (Tg) strain carrying copies of the human c-Ha-ras proto-oncogene is highly susceptible to 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary carcinogenesis, possibly due to activation of the transgene, and can be used in medium-term bioassay models to test for modifying effects of estrogenic environmental compounds on tumor development. The present study was conducted to assess the influence of dietary feeding of the endocrine disruptors atrazine and nonylphenol on DMBA-induced carcinogenesis in c-Ha-ras Tg rats. Animals of both sexes were given a single oral dose of DMBA (25 mg/kg body weight) at 50 days of age and thereafter received soybean-free diet containing 5, 50 or 500 ppm atrazine, or 10, 25, 100 or 250 ppm nonylphenol. In female Tg rats, atrazine at a dose of 5 ppm increased the incidences of mammary adenomas and adenocarcinomas (P < 0.01 and P < 0.05), while 50 ppm increased the adenocarcinoma incidence (P < 0.05). In males, skin tumor development, in contrast, was significantly decreased at the highest dose. Nonylphenol at 10 ppm increased adenocarcinoma and total mammary tumor multiplicity in female Tg rats (P < 0.05), but there was no dose dependence, a significant quadratic dose-response trend rather being observed (P < 0.05). In vitro, atrazine did not cause proliferation of MCF-7 cells at any of a range of doses tested. These results suggest that endocrine disruptors may enhance mammary carcinogenesis, but only in a certain limited dose range under the present experimental conditions. The doses applied, moreover, were all extremely high compared to the possible environmental human exposure levels. PMID- 15132768 TI - Identification of differentially expressed molecules in adult T-cell leukemia cells proliferating in vivo. AB - HTLV-I is the causative agent of adult T-cell leukemia (ATL). However, the precise mechanism underlying the neoplastic cell growth of ATL remains unclear. In this study, we established a leukemic cell line, termed SYK-11L(+), from tumor cells (S-YU) in an in vivo cell proliferation model of ATL using severe combined immunodeficiency (SCID) mice. Unexpectedly, SYK-11L(+) was found to have no tumorigenicity in SCID mice. Flow cytometric analysis showed that S-YU expressed cell adhesion molecules including CD44, ICAM-1 and OX40, whereas SYK-11L(+) had lost the expression of these molecules. The administration of anti-OX40 monoclonal antibody inhibited the engraftment of S-YU cells into SCID mice, suggesting that OX40 is a potential target for immunotherapy. Significant differences in responsiveness to IL-2 and IL-15 were observed between the two cell types. To better understand the molecular basis of tumorigenicity, cDNA microarray analysis was performed using tumorigenic S-YU and non-tumorigenic SYK 11L(+) cells. We obtained several candidate genes differentially overexpressed in S-YU compared with SYK-11L(+). Interestingly, one such gene, regulator of G protein signaling 1 (RGS1), was shown to be overexpressed in most ATL patients. Further characterization of the differentially expressed molecules, such as OX40 and RGS1, would provide useful information not only to elucidate the mechanism of ATL cell growth in vivo, but also to develop novel molecularly targeted therapies. PMID- 15132769 TI - Expression of the c-myc gene as a predictor of chemotherapy response and a prognostic factor in patients with ovarian cancer. AB - The present study was conducted to determine whether and how expression of the c myc gene is related to the response to chemotherapy in patients with epithelial ovarian cancer. This study includes 101 consecutive patients with stage Ic to IV epithelial ovarian cancer who underwent primary surgery followed by platinum based chemotherapy. Immunohistochemical studies were performed to detect Ki-67 and ARF proteins. Apoptotic cells were identified by the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate biotin nick-end labeling method. Mutation of the p53 gene was screened by polymerase chain reaction (PCR)-single strand conformation polymorphism analysis and confirmed by direct sequencing. mRNA expression of c-myc was determined by means of reverse transcription-PCR. Apoptotic index (AI) and ARF labeling index (LI) were significantly increased and Ki-67 LI was decreased after chemotherapy in patients from whom specimens could be obtained before and after chemotherapy. AI, ARF LI, and Ki-67 LI were not related to p53 gene status. A significant correlation between expression of c-myc and ARF LI was observed. Of 38 patients with measurable lesion, 23 (60.5%) responded to chemotherapy and 15 (39.5%) did not. Tumors with the wild-type p53 gene responded significantly better to chemotherapy than did tumors with the mutation. Responders showed a higher expression of c-myc than nonresponders (468 +/- 76 vs. 187 +/- 68). The receiver operating characteristic (ROC) curve according to chemoresponse demonstrated that the cut off value of c-myc expression was 200. Patients with c-myc expression of more than 200 had a better 5-year survival rate (69.8% vs. 43.5%; 101 patients). Multivariate analysis revealed that c-myc expression was an independent prognostic factor. Our results suggest that the expression of c-myc gene is related to chemoresponse and might be a useful prognostic factor in patients with epithelial ovarian cancer. PMID- 15132770 TI - Targeted gene delivery using humanized single-chain antibody with negatively charged oligopeptide tail. AB - We have recently developed the so-called recombinant immunoporter as a non-viral vector based on a single-chain antibody (scFv) derived from a monoclonal antibody B4G7 against epidermal growth factor (EGF) receptor. This immunoporter (mBBD20) was composed of single-chain antibody and negatively charged oligopeptide tail [5 units of Asn4Ser (D20)], and was expressed in yeast as a secreted protein. The purified mBBD20 was converted to an immunogene by mixing it with DNA and a cationic polymer, polyethyleneimine (PEI). The resulting complex, namely recombinant immunogene, exhibited gene transfer activity with EGFR-specificity in vitro (Suzuki et al., Gene Ther. 2003). In this paper, we further improved various conditions necessary for the formation of the proper recombinant immunogene, retaining receptor specificity of its binding, intracellular processing of the receptor-bound gene, and efficient gene expression. Moreover, we provided evidence that the recombinant immunoporter made with humanized scFv could be used as a potent gene transfer vehicle to target particular tumor cells. This approach seems worthy of clinical trial. PMID- 15132771 TI - Identification of NOL8, a nucleolar protein containing an RNA recognition motif (RRM), which was overexpressed in diffuse-type gastric cancer. AB - In an attempt to identify novel therapeutic targets for diffuse-type gastric cancer, we had previously compared expression profiles of 20 diffuse-type gastric cancer tissues with corresponding non-cancerous mucosae by means of a cDNA microarray consisting of 23,040 genes. Among 153 genes whose expression levels were elevated in cancers compared to non-cancerous mucosae, we focused on a gene termed NOL8 that encodes a putative 150-kDa protein with an RNA-recognition motif (RRM) domain in its amino-acid terminal region. Comparison of expression profiles between diffuse-type and intestinal-type gastric cancers showed that NOL8 was specifically up-regulated in diffuse-type cancers. Northern blot analysis revealed that NOL8 was expressed in skeletal muscle, but not expressed or hardly detectable in 22 other tissues examined. Immunocytochemical staining of NOL8 showed specific localization in the nucleolus. Subsequent protein phosphatase analysis coupled with western analysis revealed the presence of the phosphorylated form. Furthermore, transfection of short-interfering RNA (siRNA) specific to NOL8 into three diffuse-type gastric cancer cells, St-4, MKN45 and TMK-1, effectively reduced expression of this gene and induced apoptosis in these cells. These findings provide a new insight into diffuse-type gastric carcinogenesis and may contribute to the development of new therapeutic strategies for diffuse-type gastric cancer. PMID- 15132772 TI - Genetic link between p53 and genes required for formation of the zonula adherens junction. AB - Ectopic expression of human p53 in Drosophila eye imaginal disc cells induces apoptosis and results in a rough eye phenotype in the adult flies. We have screened Drosophila stocks to identify mutations that enhance or suppress the p53 induced rough eye phenotype. One of the dominant enhancers of the p53-induced rough eye phenotype corresponds to a loss-of-function mutation of the crumbs gene, which is essential for the biogenesis of the zonula adherens junction and the establishment of apical polarity in epithelial cells. Enhancement of p53 induced apoptosis in the eye imaginal discs by a half-reduction of the crumbs gene dose was confirmed by a TUNEL method. Furthermore, mutations of genes for Shotgun (Drosophila E-cadherin) and Armadillo (Drosophila beta-catenin), the two main components of the adherens junction, also strongly enhanced the p53-induced rough eye phenotype. These results suggest that human p53 senses subtle abnormality at the adherens junction or in signals derived from the junction, and consequently induces apoptosis to remove abnormal cells from tissue. Thus p53 likely plays a role as a guardian of the tissue not only by sensing the damaged DNA, but also by sensing signals from the adherens junction. PMID- 15132773 TI - Conditional gene silencing utilizing the lac repressor reveals a role of SHP-2 in cagA-positive Helicobacter pylori pathogenicity. AB - RNA interference (RNAi) is a newly described biological phenomenon mediated by small interfering RNA (siRNA) that targets mRNA for degradation by cellular enzymes and has become a powerful method for studying gene functions in mammalian systems. The development of systems for inducing siRNA expression should enable examination of acute loss-of-function phenotypes in a cell of interest without the need to consider lethality or epigenetic adaptation of cells. We describe in this report an inducible siRNA expression system made by combined utilization of the RNA polymerase III-dependent promoter H1 and the bacterial lac repressor. Using this system, we established AGS gastric epithelial cells in which expression of SHP-2, a cellular tyrosine phosphatase known to specifically bind the Helicobacter pylori virulence factor CagA, is conditionally and reversibly silenced by the lactose analog isopropyl-1-thio-beta-D-galactopyranoside (IPTG). Upon expression in AGS cells, CagA provoked a morphological transformation, termed the hummingbird phenotype, which is associated with CagA virulence. This morphogenetic activity of CagA was totally abolished when SHP-2 expression was silenced by inducible siRNA expression in AGS cells. Our results indicate that SHP-2 is a critical downstream effector of H. pylori CagA. The conditional gene silencing system described here should become a powerful tool for investigating the roles of cancer-related genes through a reversed genetic approach. PMID- 15132774 TI - Isoliquiritigenin, a flavonoid from licorice, reduces prostaglandin E2 and nitric oxide, causes apoptosis, and suppresses aberrant crypt foci development. AB - Isoliquiritigenin (ILTG), a flavonoid group compound, exists in some foodstuffs and herbal medicines such as licorice (Glycyrrhiza uralensis Fisher). Previously, we showed that ILTG can suppress azoxymethane (AOM)-induced colon carcinogenesis in ddY mice. In the present report, we present evidence that ILTG markedly decreases both prostaglandin E2 (PGE2) and nitric oxide (NO) production in RAW264.7 mouse macrophage cells. The decrease of PGE2 was dependent on cyclooxygenase-2 (COX-2) expression and the decrease of NO appeared due to a decrease in inducible nitric oxide synthase (iNOS) protein expression. In mouse and human colon carcinoma cells, ILTG treatment suppressed cell growth and caused apoptosis. Furthermore, in vivo administration of ILTG inhibited the induction of preneoplastic aberrant crypt foci (ACF) in the male F344 rat colon. Our results suggest that ILTG is a promising chemopreventive agent against colon carcinogenesis. PMID- 15132775 TI - DNA intrastrand cross-link at the 5'-GA-3' sequence formed by busulfan and its role in the cytotoxic effect. AB - Busulfan (1,4-butanediol dimethanesulfonate) has been used widely for the treatment of patients with chronic myelogenous leukemia. Busulfan is bifunctional and thus may effectively induce DNA damage, which may play an important role in the cytotoxicity. In this study, we compared the cytotoxicity of bifunctional busulfan with that of monofunctional ethyl methanesulfonate (EMS) in human promyelocytic leukemia HL-60 cells. Busulfan showed a significant inhibitory effect on cell growth, whereas the cells grew in the presence of EMS. To clarify the mechanism of cytotoxicity of busulfan, we investigated DNA damage induced by busulfan using 32P-5'-end-labeled DNA fragments obtained from the human p16 tumor suppressor gene. Busulfan induced DNA damage dose-dependently, whereas EMS caused little DNA damage. DNA-sequencing experiments using piperidine and 3 methyladenine DNA glycosylase indicated that busulfan caused double-base lesions mainly at 5'-GA-3' and, to a lesser extent, at 5'-GG-3' sequences. Time of flight mass spectrometry confirmed that busulfan forms an intrastrand cross-link at the 5'-GA-3' sequence, in addition to mono-alkylation. The mechanism and the role of cross-linking at the 5'-GA-3' sequence are discussed in relation to the cytotoxicity induced by busulfan. PMID- 15132776 TI - Late resistance to adenoviral p53-mediated apoptosis caused by decreased expression of Coxsackie-adenovirus receptors in human lung cancer cells. AB - Adenovirus-mediated wild-type p53 gene transfer induces apoptosis in a variety of human cancer cells. Although clinical trials have demonstrated that a replication deficient recombinant adenovirus expressing the wild-type p53 gene (Ad-p53) is effective in suppressing growth of non-small cell lung cancer (NSCLC), we often experienced late resistance to this treatment. To elucidate the mechanism of late resistance to Ad-p53 in human lung cancer cells, we generated 5 different resistant variants from p53-susceptible H1299 NSCLC cells by repeated infections with Ad-p53. We first examined the transduction efficiency of adenoviral vector by Ad-LacZ transduction followed by X-gal staining in parental and 5 resistant H1299 cell lines. Their sensitivity to viral infection decreased in correlation with the magnitude of resistance, and Ad-p53-mediated tumor suppression could be restored by dose escalation of Ad-p53 in the resistant variants. The expression of Coxsackie and adenovirus receptor (CAR) and alphaV integrins, which are cellular receptors for attachment and internalization of the virus, respectively, was next investigated in these cell lines. Flow cytometry revealed that alphaVbeta3 and alphaVbeta5 integrin expression was consistent, while p53 resistant cell lines showed that diminished CAR expression correlated with the magnitude of the resistance. Our results demonstrated that decreased CAR expression could be one of the mechanisms of late resistance to Ad-p53, which may have a significant impact on the outcome of adenovirus-based cancer gene therapy. PMID- 15132777 TI - The expressions of p21 and pRB may be good indicators for the sensitivity of esophageal squamous cell cancers to CPT-11: Cell proliferation activity correlates with the effect of CPT-11. AB - Previously, we demonstrated that CPT-11 is an effective agent against esophageal squamous cell cancers (ESCC), and that the protein level of DNA topoisomerase I can be a predictor for sensitivity to CPT-11 (Jpn J Cancer Res 2001; 92: 1335 41). Here, we describe our search for additional predictors of sensitivity to CPT 11, mainly among cell cycle-regulating proteins, because the cytotoxicity of CPT 11 is significantly correlated with the percentage of ESCC cells in S-phase. To this end, we selected and examined the expressions of 5 proteins involved in G1-S transition, i.e., p53, cyclin D1, p21, p27, and pRB, in 14 ESCC cell lines by western blot analysis. Among these proteins, the expression levels of p21 and pRB showed significant differences that were associated with the IC50 values for CPT 11 (P = 0.0339 and P = 0.0109, respectively). Namely, the expression of p21 or pRB independently could be a good indicator of CPT-11 efficacy in ESCC. In addition, the cell proliferation activities examined by enzyme-linked immunosorbent assay (ELISA) using 5-bromo-2'-deoxyuridine (BrdU) showed a significant correlation with the percentage of total S-phase cells (correlation coefficient = 0.568, P = 0.0324), and an inverse correlation with the IC50 values for CPT-11 (correlation coefficient =-0.601, P = 0.0213). Because, as in the case of DNA topoisomerase I, the cell proliferation activity determined using BrdU shows a close relationship with the MIB-1 labeling index, immunohistochemical studies of p21, pRB, and MIB-1 in resected ESCC specimens and/or biopsy samples could make it possible to predict more precisely the sensitivity of ESCC patients to CPT-11 prior to treatment. PMID- 15132786 TI - [Effects of lung protective ventilation on inhibiting inflammatory mediators released into plasma and bronchial alveolar lavage fluid in acute respiratory distress syndrome caused by pulmonary and extrapulmonary insults in dog]. AB - OBJECTIVE: To observe the changes in oxygenation index and inflammatory mediators in plasma and bronchial alveolar lavage fluid (BALF) of different lung areas (upper lobe, heart lobe, diaphragm lobe) in acute respiratory distress syndrome (ARDS) caused by pulmonary and extra-pulmonary insults with lung protective ventilation treatment in dog. METHODS: Twenty-four male mongrel dogs were randomly divided into ARDSp (ARDS caused by pulmonary disease) experimental group, ARDSp control group, ARDSexp (ARDS caused by extra-pulmonary disease) experimental group, and ARDSexp control group. In ARDSp dogs detergent was introduced intratracheally to cause lung injury, while in ARDSexp dog's oleic acid was given intravenously to produce lung injury. After lung injury, the dogs in the experimental groups received lung protective ventilation treatment (tidal volume: 8 ml/kg, positive end-expiratory pressure (PEEP): 10 cm H2O (1 cm H2O=0.098 kPa)), and the control groups received large tidal volume ventilation (tidal volume: 14-17 ml/kg, PEEP: 0). The contents of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL- 1beta), IL-6 in plasma and BALF from different areas of dog's lung (upper lobe, heart lobe, and diaphragm lobe), and arterial blood gas under lung protective ventilation treatment were measured. RESULTS: After lung injury, the results of arterial oxygenation index were getting worse, and the contents of TNF-alpha, IL-1beta, and IL-6 in plasma were obviously elevated. The contents of inflammatory mediators in upper lobe and heart lobe of ARDSp dogs were higher than that of ARDSexp dogs (all P<0.05). After receiving lung protective ventilation, the symptoms in the dogs of experimental groups were gradually getting ameliorated compared to control groups, but ARDSexp experimental dogs rallied better than ARDSp experimental dogs. CONCLUSION: There are statistical differences in the amount of inflammatory mediators released in BALF from different lung areas and arterial oxygenation amelioration between ARDSp dogs and ARDSexp dogs, and better effects are seen in ARDSexp dogs than ARDSp dogs under lung protective ventilation treatment. PMID- 15132787 TI - [Study on the damage of liver in patients with SARS]. AB - OBJECTIVE: To study the changes in liver function and histopathology, and investigate the underlying mechanism and clinical significance of damage of liver in patients with severe acute respiratory syndrome (SARS). METHODS: According the clinical diagnostic standard of atypical pneumonia of Ministry of Health P. R. China, liver function was assessed in 110 SARS patients admitted from February 2003 to June 2003. Of them 8 SARS patients died, and the livers were pathologically examined, and their liver function parameters were compared with that of the 35 healthy controls. RESULTS: Alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), total bilirubin (TBil) of patients with SARS were higher than those of controls, they were (91.61+/-50.53) U/L vs. (32.91+/-10.56) U/L, (78.68+/-33.32) U/L vs. (29.43+/ 8.89) U/L, (429.95+/-188.94) U/L vs. (200.83+/-44.86) U/L, (11.67+/-4.26) micromol/L vs. (8.44+/-3.86) micromol/L, all P<0.001. Albumin (ALB) and pro albumin (PAB) of patients with SARS were lower than those of controls, they were (34.40+/-5.13) g/L vs. (42.09+/-6.79) g/L, (0.20+/-0.06) g/L vs. (0.34+/-0.05) g/L, both P<0.001. Direct reaction bilirubin (DBil), total bile acid (TBA), gamma glutamyltransferase (GGT) and alkaline phosphatase (ALP) showed no marked difference between SARS patients and controls, all P>0.05. Non-specific inflammation in the liver was observed in pathological examination in 4 cases. ALT, AST, GGT and LDH were always 4 to 9 times of normal. The ratio of abnormality of ALT, AST and PAB were more than 80.0 percent, the ratio of abnormality of ALB was 42.7 percent, and less than 30.0 percent for other indexes. The average of LDH, ALT, and AST of dead patients were higher than those of the survivors. Histopathology of liver was non-specific hepatitis. CONCLUSION: The patients with SARS are prone to have mild non-specific hepatitis. It seldom causes the typical symptoms of hepatitis and it is easy to be ignored in clinic. PMID- 15132789 TI - [Association between a genomic polymorphism within the CD14 locus and severe sepsis susceptibility as well as prognosis in patients after extensive burns]. AB - OBJECTIVE: To investigate the relation of a lipopolysaccharide receptor CD14C 159T gene polymorphism to severe sepsis susceptibility and prognosis in patients with extensive burns. METHODS: The study group consisted of 118 normal controls and 16 patients with burns covering more than 60 percent total body surface area. Typing of each patient for the CD14C-159T gene polymorphism was performed by analyzing restriction fragments of a Hae III-digested DNA fragment obtained using polymerase chain reaction (PCR-RFLP). Genotypes were then related to the susceptibility and mortality rate of severe sepsis. RESULTS: The overall allele frequency (C 43.7 percent, T 56.3 percent) and genotype distribution (C homozygous 12.5 percent, C/T 62.5 percent, T homozygous 25.0 percent) were in agreement with the distribution in healthy volunteers. Genotype distribution in patients with severe sepsis was different from that in patients with an uncomplicated clinical course. Development of severe sepsis was increased in patients homozygous for the allele T (71.4 percent) than that of the non- sepsis patients (44.4 percent). CONCLUSION: The single base pair polymorphism at position-159 in the CD14 gene promoter might influence the development of severe sepsis in patients with extensive burns. PMID- 15132790 TI - [Risk factors scoring system for predicting acute respiratory failure after general thoracic surgery]. AB - OBJECTIVE: To establish a scoring system for predicting the occurrence of postoperative acute respiratory failure (ARF) after general thoracic surgery. METHODS: Eighty-five cases of post-operative ARF and 300 postoperative cases without ARF, treated from July 1981 to December 1998, were retrospectively analyzed by means of Logistic regression. The main risk factors were determined, and scores were allocated to the corresponding interval respectively, so as to establish the scoring system. RESULTS: Ten items were included in the system: lung function (existence of symptom of shortness of break, forced expiratory volume in one second (FEV1)), arterial blood gas index (arterial oxygen pressure (PaO2)), nutritional status, cigarette-smoking index, impact of operation (blood loss, influence on the lung), complicating diseases (asthma, myasthenia gravis), and age. The sensitivity and specificity of this system was 67.05 percent and 90.33 percent respectively. CONCLUSION: This newly established scoring system for predicting postoperative ARF has the advantage of convenience and objectivity. PMID- 15132791 TI - [Analysis of relation between the usage of corticosteroid in treatment and arthralgia as a sequela of SARS patients]. AB - OBJECTIVE: To analyze relation between the use of corticosteroids in the treatment for severe acute respiratory syndrome (SARS) patients and arthralgia as a sequela. METHODS: Clinical date of 30 SARS patients without other diseases in whom corticosteroid was used were reviewed including total dosage, duration of use, the highest dosage and its duration, and speed of reduction in dosage. The information about arthralgia was investigated one month after discharge of SARS patients from the hospital. RESULTS: The average total dosage of methylprednisolone was (4 244.16+/-2 292.30) mg, and the duration of use of the treatment was (25.36+/-5.88) days (ranging from 12 to 35 days). The maximum dosage was (321.33+/-174.03) mg/d, and the duration of its use was (7.73+/-4.08) days. The speed of reduction of dosage of corticosteroids was (21.33+/-10.18) mg/d. There were 26 of 30 patients (86.67 percent) experienced arthralgia symptom during convalescence. In 3.6 percent of patients arthralgia occurred within one month after SARS, 53.85 percent of the patients experienced low-grade arthralgia. By unifactor analysis, the total dosage and its duration of use, the highest dosage and its duration, speed of reduction of dosage of corticosteroids were correlated with the degree of arthralgia, respectively. The duration of arthralgia was correlated with the total dosage, the duration of high dosage, and high dosage. Age was not correlated with either the degree or the persisting time of arthralgia. The degree of arthralgia was only correlated with the total dosage, and the duration of arthralgia was correlated with administration time of glucocorticosteroids by multifactor analysis. CONCLUSION: There is a dosage- effect relation between the degree of arthralgia and the total dosage of corticosteroid, and a time-effect relation between the duration of arthralgia and length of the use of corticosteroids. PMID- 15132792 TI - [Clinical analysis of complications after non- invasive positive pressure ventilation and an inquiry into the respiratory treatment strategy in patients with SARS]. AB - OBJECTIVE: To analyze the clinical data of severe acute respiratory syndrome (SARS) patients with pneumothorax and mediastinal emphysema occurring after the non-invasive positive pressure ventilation (NIPPV), and to inquire into relevant strategy in respiratory treatment in the SARS patients. METHODS: Twenty-seven serious cases of SARS undergone NIPPV were analyzed retrospectively. RESULTS: Pneumothorax and mediastinal emphysema occurred in 7 of 27 serious cases of SARS with NIPPV, and mediastinal emphysema occurred in 1 of 189 cases of SARS without mechanical ventilation (MV). The incidence of pneumothorax and mediastinal emphysema in serious cases of SARS with NIPPV was significantly higher (chi2=25.052, P<0.01) than that in cases of SARS without receiving MV. The peripheral blood oxygen saturation level was not changed significantly after reasonable adjustment of ventilation pressure and increase in oxygen concentration inhaled. Seven cases of pneumothorax and mediastinal emphysema improved gradually. CONCLUSION: The incidence of pneumothorax and mediastinal emphysema in serious SARS patients with NIPPV is significantly higher than that in SARS patients without receiving MV. This might be related to SARS related pulmonary injuries, intensive cough and high mechanical ventilation pressure. The pneumothorax and mediastinal emphysema improve gradually in all cases after reasonable adjustment of mechanical ventilative pressure. So when NIPPV is used in the treatment of serious SARS patients to improve hypoxemia, optimized mechanical ventilative pressure should be acquired in order to avoid pneumothorax and mediastinal emphysema. PMID- 15132793 TI - [Study of infusion of oxygen-enriched liquid to correct severe hypoxemia in infectious diseases: a report of pilot clinical study]. AB - OBJECTIVE: To investigate a new therapy for effectively correcting severe hypoxemia in patients with infectious diseases by infusion of oxygen-enriched liquid, in order to raise the partial pressure of blood oxygen without passing through pathologically damaged alveoli of such patients. METHODS: Intravenous drip with oxygen-enriched liquids was given to 6 cases suffering from severe acute respiratory syndrome (SARS), and 3 cases of acquired immune deficiency syndrome (AIDS) in the course of treatment for 1 to 5 days, 500-700 ml per day. RESULTS: For all the 9 SARS cases, their hypoxemia was gradually corrected to normal in 20 minutes' or 4 hours' intravenous drip with oxygen-enriched liquid. Respiratory rate decreased from 29-49 breath/min to 18-22 breath/min, heart rate decreased from 89-145 beats/min to 60-79 beats/min, two faint patients regained consciousness, hypoxemia was redressed, partial pressure of oxygen in artery increased from 56 mm Hg (1 mm Hg=0.133 kPa) to 87 mm Hg, saturation of oxygen increased from 0.89 to 0.96. CONCLUSION: Intravenous drip of the oxygen-enriched liquid effectively helped correct the hypoxemia of SARS and other infectious diseases cases by bypassing the diseased alveoli through which oxygen would not pass into the blood by conventional oxygen inhalation. This therapy of oxygen enriched liquid infusion could be quite life-saving in the combined treatment for SARS and other infectious diseases. PMID- 15132794 TI - [Value of non-invasive continuous hemodynamic monitoring system in the differential diagnosis of the patients with dyspnea]. AB - OBJECTIVE: To evaluate non-invasive continuous hemodynamic (IQ System) monitoring in the differential diagnosis of dyspnea. METHODS: According to the diagnosis on discharge, 48 patients diagnosed as pulmonary dyspnea were enrolled in control group and 38 patients with cardiac dyspnea were in heart failure group. Each patient underwent IQ monitoring on admission and after recovery. The difference in the diagnosis on admission and on discharge, and the difference in IQ index were analyzed. RESULTS: (1) Clinical diagnosis: 7 patients in heart failure group were missed on admission as 5 were diagnosed as pneumonia and 2 were diagnosed as chronic obstructive pulmonary disease (COPD). One patient with pneumothorax in control group was misdiagnosed as heart failure. (2) Indexes of cardiac function: base impedance (Zo), maximum value of dz/dt (dz/dt max) and Heather index (HI) of heart failure group were markedly lower than those of control group (all P<0.001). The respective values were (19.0+/-3.5) Omega vs. (28.8+/-5.5) Omega, (0.76+/-0.42) Omega/s vs. (1.40+/-0.72) Omega/s, and (7.04+/-4.25) Omega/s2 vs. (13.60+/-6.36) Omega/s2. If Zo value of patients with dyspnea was 22 omicron or less, the sensitivity in diagnosing heart failure was 79 percent, and its specificity was 94 percent. If Zo value was 18.0 omicron or less, the sensitivity in diagnosing heart failure was 47 percent, and its specificity was 100 percent. (3) Comparison within groups: Indexes of cardiac function of control group did not change obviously and Zo, dz/dt max, HI, stroke volume (SV) and acceleration contraction index (ACI) values of heart failure group rose significantly after recovery. (4) Pre-ejection period (PEP) and left ventricular ejection time (VET) in both groups had no statistical significance in differences. CONCLUSION: IQ System was valuable in differential diagnosis to judge if dyspnea is caused by heart failure. Zo, dz/dt max and HI, especially Zo, are reliable. PMID- 15132795 TI - [Clinical effect of treatment with lipo-prostaglandin E1 on the patients with chronic glomerulonephritis]. AB - OBJECTIVE: To investigate potential mechanism underlying lipo-prostaglandin E1 (PGE1) in treatment of patients with chronic glomerulonephritis. METHODS: The recommended dose of 20 microg lipo-PGE1 for treatment of chronic glomerulonephritis was as a daily dose by continuous intravenous infusion or intravenous injection for 4 weeks. The levels of interleukin-1 (IL- 1), tumor necrosis factor-alpha (TNF-alpha), blood urea nitrogen (BUN), serum creatinine (SCr) and clearance of creatinine (CCr) were measured before and after treatment. RESULTS: After treatment with lipo-PGE1, levels of IL-1, TNF-alpha, BUN and SCr were lower than those before treatment (all P<0.01), but CCr values were higher than those before treatment (P<0.05). CONCLUSION: Lipo-PGE1 can reduce the renal inflammatory response and improve the renal function in the patients with chronic glomerulonephritis. PMID- 15132798 TI - [Clinical characteristics and outcome of adult diabetic patients with ketosis as the initial manifestation: a follow-up study]. AB - OBJECTIVE: To study the clinical characteristics and natural history of adult diabetic patients with ketosis as the initial manifestation. METHODS: In 60 patients with ketosis as the initial symptom, the mean age was (42.78+/-11.14) years and body mass index(BMI) was (2.34+/-3.47) kg/m2. The islet cell antibody (ICA), anti-glutamic acid decarboxylase (GAD) antibody, fast plasma glucose and insulin level, glucose and insulin level of 2 hours after oral glucose test were measured. After insulin therapy, these patients received therapy plan according to their glucose level. During follow-up period, these items were tested again. RESULTS: There were 27 patients using insulin and 33 patients using diet or oral hypoglycemic drugs for controlling blood glucose during the follow-up. Compared with patients using insulin therapy, patients using oral hypoglycemic drugs or diet therapy showed higher BMI (at onset of ketosis (25.69+/-2.66) kg/m2 vs. (22.64+/-3.75) kg/m2, P=0.001; during follow-up period (26.19+/-3.11) kg/m2 vs. (22.29+/-2.97) kg/m2, P=0.000), higher post-load serum insulin level (at onset of ketosis (32.39+/-35.87) mU/L vs. (18.53+/-14.43) mU/L, P=0.017; during follow-up period (67.34+/-68.42) mU/L vs. (36.40+/-45.39) mU/L, P=0.004), lower HbA 1C (at onset of ketosis (10.76+/-1.25) percent vs. (12.04+/-1.99) percent, P=0.006; during follow-up period (6.97+/-1.31)percent vs. (9.36+/-3.18) percent, P=0.000), higher percentage of ICA and GAD positivity (at onset of ketosis, ICA positive rate was 18.52 percent vs. 0, P=0.030, GAD positive rate was 18.52 percent vs. 0, P=0.030; during follow-up period, ICA positive rate was 7.41 percent vs. 0, P=0.111, GAD positive rate was 14.81 percent vs. 0, P=0.022). CONCLUSION: Among the adult diabetic patients with ketosis as the first symptom, quite a number of patients are not insulin dependent for the long-term blood sugar control. Ketosis is not a good index for differentiating type 1 from type 2 diabetes. PMID- 15132797 TI - [Diagnostic effect of bronchoalveolar lavage in early lung injury caused by enterogenic infection in rat]. AB - OBJECTIVE: To study the diagnostic effect of bronchoalveolar lavage in early lung injury by observing changes in inflammatory mediators in early lung injury caused by enterogenic infection. METHODS: Eighty-four Sprague-Dawley rats were randomly divided into infection group and sham-operation group. Cecal ligation and perforation was utilized to produce abdominal infection in rats. Six groups were sacrificed respectively at 0, 24, 48, 72, 96, 120 hours after operation. The differential cell count in bronchoalveolar lavage fluid (BALF) was assessed. The concentrations of endotoxin, phospholipase A2 (PLA2) and tumor necrosis factor alpha (TNF-alpha) in BALF, lung and plasma were assayed. RESULTS: The neutrophil percentage of BALF increased progressively. The concentrations of endotoxin, PLA2 and TNF-alpha in BALF, lung and plasma were significantly increased. The levels of endotoxin and PLA2 in lung tissue were respectively correlated positively with those in BALF and plasma (BALF and lung: r=0.904, P<0.05; BALF and plasma: r=0.895, P<0.05; lung and plasma: r=0.946, P<0.01). Significant positive correlation was also present between the TNF-alpha levels in BALF and lung (r=0.952 P<0.01), but not between the TNF-alpha level in plasma and that in lung or BALF (r=0.684, r=0.608, both P>0.05). CONCLUSION: The examinations of bronchoalveolar lavage may help discover early lung injury caused by enterogenic infection. PMID- 15132811 TI - Development of clinical practice guidelines: evaluation of 2 methods. AB - The aim of this study was to compare 2 methods for developing a clinical practice guideline (CPG) on the management of asymptomatic, impacted mandibular third molars. Outcome measures were the mean time invested by the participants for each method, the quality of the CPGs measured using the Appraisal of Guidelines for Research and Evaluation (AGREE) indicator and observations of the group discussions. We used a national consensus procedure following the Rand modified Delphi procedure (2 panels) and a local consensus procedure (2 existing dental peer groups). The mean time spent was about equal for the 2 methods. The quality of the CPGs developed by the expert panels was higher than that of the CPGs developed by the dental peer groups. Observation indicated that all group processes were influenced by the chairperson. We concluded that the expert panel method is suitable for developing reliable CPGs on a national or regional level. PMID- 15132812 TI - Cancer-related oral health care services and resources: a survey of oral and dental care in Canadian cancer centres. AB - PURPOSE: Prevention and management of oral complications of cancer and cancer therapy will improve oral function and quality of life, and reduce morbidity and the cost of care. Oral assessment, and oral and dental care have been strongly recommended before cancer therapy and should be continued during and after cancer therapy. The purpose of this survey was to assess the resources available for oral care in Canadian cancer centres. METHODS: Provincial cancer centres were assessed by questionnaire to determine the resources available for oral care in these facilities. RESULTS: Wide variability in oral and dental care of patients with cancer across Canada and a lack of documented standards of care were reported. Very few cancer centres had institutionally supported dental staff to support the oral care of patients with cancer, and few had dental treatment capability on site. The majority of centres managed oral care needs in the community with the patient's prior dentist. CONCLUSIONS: We recommend that national guidelines be developed for medically necessary oral and dental care for patients with cancer. PMID- 15132813 TI - Upper body musculoskeletal symptoms in Sardinian dental students. AB - PURPOSE: The purpose of this study was to determine how early in a dental career musculoskeletal problems develop, by comparing the prevalence of such symptoms in a dental student population in Sardinia (Italy), a control sample of psychology students from the same university and a sample of dental students in Lebanon. METHODS: One hundred and fourteen dental students from the University of Cagliari (Cagliari, Sardinia, Italy) were surveyed by questionnaire about the presence of several upper body symptoms. Their responses were compared with those of 2 different populations: 114 psychology students from the same university, matched for age and sex, and 178 dental students from the University of Saint Joseph (Beirut, Lebanon). The psychology students were chosen for comparison because of the general dissimilarity of their daily activity to that of dentists. RESULTS: The Italian and Lebanese dental students did not differ with regard to presence of symptoms. The Italian dental students differed from the psychology students in just one respect, with the dental students reporting more lower back pain. Headache was the most prevalent symptom in all 3 groups and was more prevalent among women; however, this difference between the sexes did not reach statistical significance for the psychology students. CONCLUSIONS: The appearance of musculoskeletal symptoms among dental students, even after a relatively short clinical training period, suggests that ergonomics should be covered in the educational system to reduce risks to dental practitioners. PMID- 15132814 TI - A closer look at diagnosis in clinical dental practice: part 2. Using predictive values and receiver operating characteristics in assessing diagnostic accuracy. AB - When a clinician is planning to use a diagnostic test or procedure, it is important to establish the likelihood that an individual patient is affected by the condition or disease; this determination depends on predictions that are affected by various features of the diagnostic procedure. In this regard, sensitivity and specificity are limited because they describe the results of a procedure in a dichotomous way: the result is either positive or negative. However, many clinical procedures are not dichotomous, such as probing of periodontal pockets or assessment of radiographs for caries, and in these situations, a range of features is examined to produce a degree of certainty regarding the presence or absence of disease. This article examines predictive values and receiver operating characteristic (ROC) analysis, an algorithm that combines various statistical features of diagnostic procedures to assess the effectiveness of nondichotomous procedures without imposing an arbitrary threshold. PMID- 15132815 TI - Mucocele of the upper lip: case report of an uncommon presentation and its differential diagnosis. AB - This report describes a lesion of the upper lip that was definitively diagnosed by histologic examination as a mucocele or mucus retention phenomenon. The usual location of mucoceles is the lower lip. This case illustrates an uncommon presentation of mucocele with respect to symptoms, location and duration. The features of a variety of oral lesions are discussed and compared, to help clinicians in establishing an appropriate differential diagnosis. PMID- 15132817 TI - Frequent ventricular premature beats increase blood pressure variability in rats. AB - AIM: The present study was designed to test a hypothesis that nonfatal ventricular arrhythmia such as ventricular premature beats (VPB) is a contributing factor in the elevation of blood pressure variability (BPV). METHODS: Blood pressure (BP) and electrocardiogram were continuously recorded. The relation between VPB and BPV was observed under conscious state in chronic myocardial infarction (MI) rats one month after ligation of the left coronary artery, and further verified under anesthetized state in rat model of ventricular arrhythmia produced by acute intravenous infusion of aconitine. RESULTS: MI rats exhibited a big difference in the count and pattern of VPB, and were divided into no VPB, occasional VPB, and frequent VPB groups. Among the three groups, there were no differences in BP, heart period (HP), and MI size. However, BPV was markedly higher in frequent not occasional VPB rats, and HP variability (HPV) was larger in both frequent and occasional VPB rats, when compared with no VPB rats. In the whole population of MI rats, BPV was positively correlated with VPB and HPV, not with BP, HP and MI size. Infusion of aconitine had no effect on BP, HP, BPV, and HPV during the period without VPB. Frequent VPB after several minutes of aconitine infusion induced significant increase in BPV and HPV with no change in BP and HP. BPV was also positively correlated with VPB and HPV, not with BP and HP. Hemodynamics in aconitine-evoked ventricular tachycardia was characterized as lower BP, higher BPV, and higher HPV. CONCLUSION: High BPV can be caused by frequent not occasional VPB in rats. PMID- 15132816 TI - Hardening of dual-cure resin cements and a resin composite restorative cured with QTH and LED curing units. AB - OBJECTIVE: The aim of this study was to determine the effects of light intensity and type of light unit (quartztungsten-halogen [QTH] or light-emitting diode [LED]) on the hardening of various resin cements and a resin composite restorative. METHODS: Disk specimens were prepared from 4 dual-cured resin cements (Variolink II, Calibra, Nexus 2 and RelyX ARC). Two QTH light-curing units (Visilux 2, at 550 mW/cm2, and Optilux 501, at 1,360 mW/cm2) and a LED unit (Elipar FreeLight, at 320 mW/cm2) were used for curing. Specimens were light cured or dual-cured for 10, 30 or 40 seconds with 1 of the 3 light units (curing applied to upper surface only) and were tested 24 hours after curing. Additional cement specimens were self-cured and tested at 15, 30 and 60 minutes and at 24 hours. Testing consisted of measurement of Knoop hardness number (KHN) for each specimen. Six KHN values were obtained for the upper surface only of the various cement specimens in each test group. Disk specimens 2.5 mm thick were also prepared from a resin composite restorative (XRV Herculite). These were light cured as above, and KHN measurements were obtained for both the upper and the lower surfaces. Mean KHNs were determined, and data were analyzed with analysis of variance. RESULTS: The groups were significantly different (p < 0.05). High intensity light curing resulted in the highest KHN values for all materials with any of the 3 light-curing times. For the cements, LED light curing (with both dual-curing and light-curing modes) resulted in hardness values similar to those achieved with conventional QTH light curing, although there were some exceptions. However, both LED and conventional QTH light curing resulted in inferior hardening of lower surfaces of the XRV Herculite specimens at the 3 curing times. For all cements except Nexus 2, self-curing resulted in significantly lower hardness values than dual curing. The self-curing mechanism of Variolink II cement needed a longer time to activate than those of the other cements. CONCLUSIONS: High-intensity light curing and longer curing times resulted in the highest KHNs. The LED curing unit was associated with the lowest hardness values for lower surfaces of the resin composite restorative. PMID- 15132818 TI - Inactivation gating determines drug potency: a common mechanism for drug blockade of HERG channels. AB - AIM: To determine the mechanisms of interactions between different drugs and HERG channels. METHODS: Various antiarrhythmic (dofetilide, quinidine, azimilide, RP58866) and non-antiarrhythmic (terfenadine, nicotine) agents were used on HERG channels expressed in Xenopus oocyte. Whole-cell voltage-clamp techniques were used. RESULTS: All drugs produced concentration-dependent block of HERG current. The inhibition was markedly facilitated with voltage protocols favoring channel inactivation (eg, less negative holding potentials). Maneuvers that weakened channel inactivation (eg, elevation of external K+), relieved HERG blockade by all drugs. Moreover, the inhibitory potency was reduced by at least 20-300 fold with varying compounds when rapid C-type inactivation was removed by a mutation located between the transmembrane domains 5 and 6 (S631A). CONCLUSION: The inactivation gating of HERG channels determines the blocking potency of drugs. This mechanism might be common to drugs of various classes. PMID- 15132819 TI - Effect of angiotensin II receptor 1 antisense oligodoexynucleotides on physiological and pathophysiological growth of cardiomyocytes. AB - AIM: To evaluate the role of angiotensin II receptor 1 antisense oligodexynucleotides (AT1R-AS-ODNs) on physiological and pathophysiological growth of cardiomyocytes from normotensive rats. METHODS: Cardiomyocytes were transfected with AT1R-AS-ODNs (200 nmol/L) followed by treatment with or without angiotensin II (1 micromol/L). In situ hybridization and Western blot were used for AT1R mRNA and protein detection, respectively. c-Jun N-terminal protein kinase (JNK) activity was characterized by immune complex kinase assay. c-Jun protein expression was examined by immunocytochemistry. DNA content was detected by flow cytometric assay. Atrial natriuretic factor (ANF) expression was identified by radioimmunoassay. RESULTS: Treatment with AT1R-AS-ODNs for 24 h resulted in 51.2 % decrease in AT1R mRNA and 60.7 % in protein (P<0.05 vs control). However, the basal level of JNK activity, c-Jun protein expression, and DNA content were not altered by AT1R-AS treatment in absence of overactive hormonal system. After treatment with angiotensin II for 30 min, both p46JNK and p54JNK were robustly activated. By 2 h, c-Jun protein expression was increased. By 24 h, angiotensin II caused a marked increase both in G0/G1 and G2/M DNA content, and increased ANF expression by 1.8-fold. All these were inhibited by AT1R-AS-ODNs pretreatment. In contrast, sense sequence was ineffective. CONCLUSION: Decrease of AT1R expression by AS-ODNs did not interfere with normal growth, but protected cardiomyocytes from angiotensin II-dependent pathophysiological growth. PMID- 15132820 TI - Enhancement of naked FIX minigene expression by chloroquine in mice. AB - AIM: To study the effect of chloroquine on the expression of human clotting factor IX (hFIX) in mice. METHODS: Hydrodynamics-based naked DNA plasmid administration was performed by tail vein injection of 10 microg of pCMV- hFIX and chloroquine (0, 100, 200, and 500 micromol/L) in 2.2 mL of Ringer's solution within 6-7 s, the level and stability of hFIX expression, liver damage and toxicity were then examined. RESULTS: The maximum expression of hFIX level was 4.4+/-1.8 mg/L at 8 h after injection, 9.7+/-1.6 mg/L at 24 h only existed in 200 micromol/L chloroquine-treated animals, which is 3-4 fold higher than that of control (P<0.01). There is no significant difference observed among all the treated groups, 3 d later. Transaminase level and liver histological study showed the damage of liver was not related to chloroquine (P>0.05). CONCLUSION: Chloroquine can enhance and sustain exogenous gene expression in vivo without side effect under our experimental conditions. PMID- 15132821 TI - Effects of copper-aspirin complex on platelet-neutrophil interactions. AB - AIM: To investigate the effects of copper-aspirin complex on rat thrombosis and the interaction between platelets and neutrophils. METHODS: The model of electrically stimulated carotid artery thrombosis in Sprague Dawley rats was used; the effects of copper-aspirin complex on rat platelet-neutrophil adhesion and platelet aggregation stimulated by activated neutrophils were observed by rosette assay and Born's method, respectively. RESULTS: Intragastric copper aspirin complex (5, 7, and 10 mg/kg) dose-dependently prolonged the occlusion time; it significantly decreased the rosette number formed between thrombin activated platelets and neutrophils; the 50 % of inhibitory concentration (IC50) was (54.6+/-4.3) micromol/L. Copper-aspirin complex markedly inhibited rat platelet aggregation induced by either cell free supernatant of activated neutrophils or by activated neutrophil suspension. The values of IC50 were (224.5+/-16.2) micromol/L and (820.5+/-21.4) micromol/L, whereas aspirin had no influence. CONCLUSION: Copper-aspirin complex inhibited platelet-neutrophil interactions through a different property from aspirin and resulted in a more potent antithrombotic activity. PMID- 15132822 TI - Oxidized LDL upregulated ATP binding cassette transporter-1 in THP-1 macrophages. AB - AIM: To study the effect of oxidized low density lipoprotein (ox-LDL) on ATP binding cassette transporter A1 (ABCA1) in THP-1 macrophages. METHODS: After exposing the cultured THP-1 macrophages to ox-LDL for different periods, cholesterol efflux was determined by FJ-2107P type liquid scintillator. ABCA1 mRNA and protein level were determined by reverse trancriptase-polymerase chain reaction (RT-PCR) and Western blot, respectively. The cholesterol level in THP-1 macrophage foam cells was detected by high performance liquid chromatography. RESULTS: ox-LDL elevated ABCA1 in both protein and mRNA levels and increased apolipoprotein (apo) A-I-mediated cholesterol efflux in a time- and dose dependent manner. 22(R)-hydroxycholesterol and 9-cis-retinoic acid did significantly increase cholesterol efflux in THP-1 macrophage foam cells (P<0.05), respectively. Both of them further promoted cholesterol efflux (P<0.01). As expected, liver X receptor (LXR) agonist decreased content of esterified cholesterol in the macrophage foam cells compared with control, whereas only a slight decrease of free cholesterol was observed. LXR activity was slightly increased by oxidized LDL by 12 % at 12 h compared with 6 h. However, LXR activity was increased about 1.8 times at 24 h, and oxidized LDL further increased LXR activity by about 2.6 times at 48 h. CONCLUSION: ABCA1 gene expression was markedly increased in cholesterol-loaded cells as a result of activation of LXR/RXR. ABCA1 plays an important role in the homeostasis of cholesterol in the macrophages. PMID- 15132823 TI - Effect of G(alphaq/11) protein and ATP-sensitive potassium channels on prostaglandin E(1) preconditioning in rat hearts. AB - AIM: To investigate the effect of G(alphaq/11) signaling pathway and ATP sensitive potassium channels (K(ATP) channels) on prostaglandin E1 (PGE1) induced early and delay-preconditioning protection in rat hearts. METHODS: Two series of experiments were performed in Wistar rat hearts. In the first series of experiment, all rats were pretreated with PGE1 40 min or 23 h 20 min before the experiment. Ischemia-reperfusion injury was induced by 30 min coronary artery occlusion followed by 90 min reperfusion. Hemodynamics, infarct size, and scores of ventricular arrhythmias were measured. The expression of G(alphaq/11) protein in the heart was measured by Western blot analysis in the second series. RESULTS: Preconditioning with PGE1 (25 microg/kg) markedly reduced infarct size, left ventricular end-diastolic pressure, and scores of ventricular arrhythmia. The effect of PGE1 was significantly attenuated by glibenclamide (1 mg/kg, ip), a nonselective K(ATP) channel inhibitor. PGE1 caused a significant increase in the expression of G(alphaq/11) protein. CONCLUSION: Activations of G(alphaq/11) signal pathway and K(ATP) channel played significant roles in the cardioprotection of PGE1 preconditioning in rat heart and might be an important mechanism of signal transduction pathway during the PGE1 preconditioning. PMID- 15132824 TI - Angiotensin II stimulates phosphorylation of 4E-binding protein 1 and p70 S6 kinase in cultured vascular smooth muscle cells. AB - AIM: To examine the regulatory effects of angiotensin II (Ang II) on the phosphorylation of 4E-binding protein 1 (4E-BP1) and p70 S6 kinase in cultured vascular smooth muscle cells (VSMC), and the contribution of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (PKB) signaling pathway in this process. METHODS: VSMC obtained from rat thoracic aortas were cultured. The phosphorylation of 4E-BP1 and p70 S6 kinase was detected by immunoblotting. RESULTS: Ang II significantly increased the phosphorylation of 4E-BP1 and p70 S6 kinase, with the peaks occurring at, respectively, 10 min and 30 min, after stimulation with Ang II. The stimulatory effect of Ang II on 4E-BP1 and p70 S6 kinase phosphorylation was abrogated by Ang II type 1 receptor (A(T1) receptor) antagonist losartan, and suppressed by PI3K inhibitor LY294002 in a concentration dependent manner. CONCLUSION: Ang II treatment of VSMC induces the phosphorylation of 4E-BP1 and p70 S6 kinase via A(T1) receptor, and PI3K signaling pathway is involved in this process. PMID- 15132825 TI - Toosendanin increases free-Ca(2+) concentration in NG108-15 cells via L-type Ca(2+) channels. AB - AIM: To examine if toosendanin (TSN) affects intracellular free-Ca(2+) concentration ([Ca(2+)](i)) in neuroblastoma pluglioma hybrid cells (NG108-15 cells). METHODS: The [Ca(2+)](i) was determined by laser-scanning confocal microscopic imaging technique in which Fluo-3 was used as Ca(2+) indicator. RESULTS: TSN induced an increase in resting [Ca(2+)](i) and in high K(+)-evoked Ca(2+) transient in differentiated NG108-15 cells. The TSN-induced increase in [Ca(2+)](i) was dose-dependent and disappeared in CdCl(2-), nifedipine-containing or Ca(2+)-free solution, and appeared after washing out the Ca(2+) channel blockers or adding Ca(2+). CONCLUSION: TSN increased [Ca(2+)](i) in differentiated NG108-15 cells. The [Ca(2+)](i) enhancement was due to the influx of extracellular Ca(2+) and related to L-type Ca(2+) channels. PMID- 15132826 TI - Effects of annexins II and V on survival of neurons and astrocytes in vitro. AB - AIM: To study the effects of annexins II and V on the survival and neurite outgrowth of primary cultured neurons and the survival of astrocytes after peroxide and hypoxia insults in vitro. METHODS: Annexins II and V proteins and/or corresponding antibodies were added to the medium of primary neocortical cultures. H(2)O(2) and NaN(3) were used to induce neuron injury, respectively. Lactate dehydrogenase (LDH) release was measured. RESULTS: Addition of annexin II or V into the culture medium did not affect the normal survival and neurite outgrowth of cortical neurons. However, when an antibody against annexin II or V was added to the culture, the survival and neurite outgrowth of these neurons markedly declined. Further, addition of the two annexins into cortical cultures after peroxide and hypoxia insults markedly reduced the LDH release and cell death. CONCLUSION: Annexins II and V are essential for the survival and neurite outgrowth of developing cortical neurons, the survival of glial cells, and protect neurons and glial cells against peroxide and hypoxia injuries. PMID- 15132827 TI - Upregulation of heparin-binding growth-associated molecule after spinal cord injury in adult rats. AB - AIM: To investigate whether traumatic spinal cord injury (SCI) induces changes of gene expression of heparin-binding growth-associated molecule (HB-GAM). METHODS: In a spinal cord transection model, HB-GAM expression and cellular localization were examined using Northern blot, RT-PCR, immunohistochemistry and immunofluorescence double-labeling methods. RESULTS: HB-GAM mRNA was significantly upregulated in spinal cord tissues rostral and caudal to the injury at 7 d after SCI. HB-GAM gene expression was markedly increased at 3 d, peaked at 7 d, and declined to the baseline level at 28 d post-injury. During its peak expression, HB-GAM was co-localized in astrocytes, oligodendrocytes, and neurons in spinal cord tissues within 7 mm from the site of spinal transection. CONCLUSION: SCI induces HB-GAM expression at both mRNA and protein levels in areas close to the injury. Both neurons and glial cells expressed HB-GAM implying that HB-GAM played a role in the process of injury and/or repair following SCI. PMID- 15132828 TI - Brain ischemia induces serine phosphorylation of neuronal nitric oxide synthase by Ca(2+)/calmodulin-dependent protein kinase II in rat hippocampus. AB - AIM: To investigate whether brain ischemia induces serine phosphorylation of neuronal nitric oxide synthase (nNOS) by Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) and the interaction between CaMKII? and nNOS in rat hippocampus. METHODS: Brain ischemia was induced by bilateral carotid artery occlusion procedure. Phosphorylation and the interaction of proteins were studied by immunoprecipitation and immunoblotting. We investigated during brain ischemia serine phosphorylation and amount of nNOS in crude membranes fraction (P) and cytosolic fraction (S), interaction between CaMKIIalpha and nNOS, and the effects of 1-[N,O-bis-(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN 62, a selective inhibitor of CaMKII) on phosphorylation and the interaction of proteins in P. RESULTS: Serine phosphorylation of nNOS in P increased persistently during brain ischemia, and 15 min ischemia-induced serine phosphorylation of nNOS was attenuated significantly by KN-62. But there was no serine phosphorylation of nNOS in S. The distributions of nNOS were not affected by ischemia and KN-62. However, the binding levels of both CaMKIIalpha with nNOS and Thr(286) autophosphorylated CaMKIIalpha with nNOS increased after ischemia, and were diminished by KN-62. CONCLUSION: CaMKII interacted with nNOS and regulated serine phosphorylation of nNOS during brain ischemia. PMID- 15132829 TI - Modulatory effect of substance P on GABA-activated currents from rat dorsal root ganglion. AB - AIM: To explore the modulatory effect of substance P (SP) on GABA-activated current of dorsal root ganglion (DRG) neurons in rat. METHODS: The whole-cell patch-clamp technique was used to record SP- and GABA-activated currents in neurons freshly dissociated from rat DRG neurons. Drugs were applied by rapid solution exchange. RESULTS: Application of SP (28/41, 68.5 %) and GABA (36/41, 88.2 %) could induce concentration-dependent inward current in some cells. SP-(10 micromol/L) and GABA (100 micromol/L)-activated inward currents were (244+/-83) pA (n=9) and (1.8+/-0.5) nA (n=13), respectively. The majority of GABA-activated current had obvious three processes, the peak value (I(p)), the steady state (I(ss)) and the desensitization (I(d)). The desensitization of GABA-activated current was a biphasic process, including fast and slow desensitization. However, pre-application of SP (0.001-1 micromol/L) could inhibit the GABA-activated inward current which was identified to be GABAA receptor-mediated current. The inhibitory effects were concentration-dependent. The inhibitory effect of SP on the peak value of GABA-activated current was more than the steady state of GABA activated current. The inhibition of GABA-activated current by SP (0.1 micromol/L) was related to the time after application of SP, the inhibition of GABA-activated currents by SP reached the peak at about 4 min (49.8 %+/-7.2 %, n=7, P<0.01) and took about 12 min to get a full recovery. The inhibition of GABA activated currents by SP was almost completely removed after blockade of PKC by H 7 with the re-patch clamp. CONCLUSION: Pre-application of SP exerts a more strong inhibitory effect on the peak value of GABA-activated current than the steady state of GABA-activated current. PMID- 15132830 TI - Activation and involvement of JNK1/2 in hydrogen peroxide-induced neurotoxicity in cultured rat cortical neurons. AB - AIM: To investigate the role of c-Jun N-terminal protein kinase 1 and 2 (JNK1/2) and the main signal pathway for its activation in hydrogen peroxide (H(2)O(2)) induced apoptotic-like cortical cell death. METHODS: Using the model of oxidative stress induced by H(2)O(2), the expression and diphosphorylation of JNK1/2 was examined by immunoblotting analysis, and neuronal apoptotic like cell death was determined by 4',6-diamidino-2-phenylindole (DAPI) staining. RESULTS: The elevation in diphosphorylation level of JNK1/2 (4.40-/5.61-fold vs sham control) was associated with the concentration of H(2)O(2) (0-100 micromol/L) and the development of apoptotic-like cell death (11.04 %-81.01 %). There was no alteration of JNK1/2 protein expression following H(2)O(2) treatment and recovery at different time points. Administration with JNK1/2 antisense oligonucleotides not only significantly decreased JNK1/2 protein expression and activation level, but also significantly reduced cortical cell death induced by H(2)O(2) exposure. Furthermore, both JNK1/2 diphosphorylation and apoptotic-like cell death were largely prevented by pretreatment with (5S,10R)-(-)-5-methyl-10,11-dihydro-5H dibenzo[a,d]cyclohepten-5,10-imine hydrogen maleate (MK-801) or omission of Ca(2+) in incubation medium with ethylene glycol-bis(2-aminoethylether)-N,N,N',N' tetraacetic acid (EGTA). CONCLUSION: JNK1/2 is activated and participates in H(2)O(2)-induced apoptotic-like death in cultured rat cortical neurons mainly via N-methyl-D-aspartate (NMDA) receptor-mediated influx of extracellular Ca(2+). PMID- 15132831 TI - Precuneus contributes to attentive control of finger movement. AB - AIM: To examine whether precuneus subserves the attentive control of finger movement or whether it mediates the movement preparation and motor inhibition. METHODS: In the Preparation Stage, subjects were shown with a 5-number string in which each number stood for a key-pressing response, the number strings included a complex pattern (eg, 4-1-4-2-3), or a simple one (eg, 2-2-2-2-2), or a null one (ie, x-x-x-x-x). In the Execution Stage, five reaction signs were presented one by one and subjects were required to press the corresponding key to each sign sequentially (eg, in the 4-1-4-2-3 preparation example, subjects press key 4 to the first sign, press key 1 to the second sign, key 4 to the third sign and so on). For the null preparation pattern, five numbers, rather than the reaction signs, were shown at the same pace as in the other two conditions and subjects were to press the corresponding keys. RESULTS: Left medial frontal gyrus (BA 6) and precentral gyrus (BA 6) were involved in both of the Preparation Stage and the Execution Stage, whereas left precuneus (BA 7) was activated only in the Execution Stage. CONCLUSION: Precuneus mediates the attentive control of finger movement, but not the movement preparation or motor inhibition. PMID- 15132832 TI - L-type calcium channel blockers enhance 5-HTP-induced antinociception in mice. AB - AIM: To investigate the involvement of L-type Ca(2+) channels in antinociceptive action induced by the 5-HT precursor, 5-hydroxytryptophan (5-HTP). METHODS: Female Kunming mice were treated with either 5-HTP (20-80 mg/kg, ip) alone, or the combination of 5-HTP and fluoxetine (2-8 mg/kg, ip), pargyline (15-60 mg/kg, ip), nimodipine (2.5-10 mg/kg, ip), nifedipine (2.5-10 mg/kg, ip), verapamil (2.5 10 mg/kg, ip), CaCl(2) (5-20 mmol/L, icv), or EGTA (0.5-3 mmol/L, icv) prior to the hot-plate test (55 degree, hind-paw licking latency). In addition, locomotor activity in mice treated with 5-HTP alone was measured using an ambulometer with five activity boxes. RESULTS: Ip injection of 5-HTP alone had no influence on the spontaneous locomotor activity, whereas dose-dependently increased the latency to licking hind-paw in the hot-plate test in mice. The inhibitory effects of 5-HTP on nociceptive response were significantly enhanced by fluoxetine in the mouse hot-plate test. At a sub-effective dose, pargyline could cause a leftward shift in the dose-response curve of 5-HTP-induced antinociception. Co-administration with 5-HTP and nimodipine, nifedipine, or verapamil obviously potentiated the antinociceptive effects elicited by 5-HTP. Interestingly, 5-HTP-induced antinociception was antagonized by CaCl(2) and enhanced by EGTA injected icv in the mouse hot-plate test. CONCLUSION: These findings suggest that systemic administration of 5-HTP may yield the antinociceptive effects, which are related to Ca(2+) influx from extracellular fluid through L-type Ca(2+) channels. PMID- 15132833 TI - Alterations in circadian rhythms of melatonin and cortisol in patients with bronchial asthma. AB - AIM: To investigate the possible relationships between alterations in circadian rhythm of melatonin, cortisol and bronchial asthma. METHODS: Salivary melatonin and cortisol were measured simultaneously by radioimmunoassay in 10 mild intermittent or persistent patients, 11 moderate-to-severe persistent asthma patients, and 15 control subjects. Twelve salivary samples were collected in a series during a 24-h period in each subject. RESULTS: The results showed overall lower levels of salivary melatonin in asthma patients compared with control subject (P<0.01). The amplitude, peak-level, and baseline of salivary melatonin were significantly lower in mild intermittent or persistent (P<0.01, P<0.05) and moderate-to-severe persistent asthma patients (P<0.01) compared with control group. The 24-h mean level of salivary cortisol was greatly lower and the acrophase was markedly delayed in patients with mild intermittent or persistent asthma (P<0.01) and moderate-to-severe persistent asthma (P< 0.05, P<0.01) compared with control subject. CONCLUSION: Disordered circadian rhythms of salivary melatonin and cortisol were found in asthma patients, which may be involved in the pathogenesis of bronchial asthma. PMID- 15132835 TI - Anticarcinogenic and antioxidant activity of diindolylmethane derivatives. AB - AIM: To investigate the synthesis methods and the bioactivity of diindolylmethane (DIM) derivatives. METHODS: 1) A 3D-Quantitative Structure-Active Relationships (QSAR) Comparative Molecular Field Analysis (CoMFA) study of 14 DIM derivatives was investigated to predict their anticarcinogenic activity. 2) Based on CoMFA model, a series of new derivatives of DIM were designed and synthesized. 3) Their free radical scavenging and antioxidant potentials were tested using in-vitro DPPH radical scavenging and ?-carotene antioxidant models. 4) The anticarcinogenic activities of some compounds were tested by using microculture tetrazolium assay (MTT) and sulforhodamine B (SRB) proteochromosomic assays. RESULTS: 1) The CoMFA model derived from DIM analogues proved a good predictive ability with q2 value of 0.827. 2) New designed compounds 3c and 4c exhibited 3 fold more potent radical scavenging activity than reference substance Vitamin E in DPPH model expressed by IC50 values. 3) The primary antitumor screening essay showed that some DIM derivatives designed exhibited the inhibitory activities to some tumor cell growth at relatively high concentration, and DIM was the most effective among them. CONCLUSION: DIM's 3D-QSAR model is reliable. According to it, eleven DIM derivatives were synthesized, and two derivatives of them possess potent radical scavenging activities and some showed the inhibitory activities in primary anticancer assay in vitro. PMID- 15132834 TI - Effects of Cordyceps militaris extract on angiogenesis and tumor growth. AB - AIM: To evaluate the effects of Cordyceps militaris extract (CME) on angiogenesis and tumor growth. METHODS: Human umbilical vein endothelial cells (HUVEC), HT1080, and B16-F10 cells were used. DNA fragment, angiogenic related gene expressions (MMPs, bFGF, VEGF, etc), capillary tube formation, wound healing in vitro, tumor growth in vivo were measured. RESULTS: CME inhibited growth of HUVECs and HT1080 (P<0.01). CME 100 and 200 mg/L reduced MMP-2 gene expression in HT1080 cells by 6.0 % and 22.9 % after 3-h and 14.9 % and 32.8 % after 6-h treatment. CME did not affect MMP-9 gene expression in B16-F10 melanoma cells. CME 100 and 200 mg/L also reduced bFGF gene expression in HUVECs by 22.2 % and 41.3 %. CME inhibited tube formation of endothelial cells in vitro and in vivo. CME repressed the growth of B16-F10 melanoma cells in mice compared with control group (P<0.05). CONCLUSION: CME has antiangiogenetic properties. PMID- 15132836 TI - Inhibitors of 5-lipoxygenase inhibit expression of intercellular adhesion molecule-1 in human melanoma cells. AB - AIM: To study the effect of 5-lipoxygenase inhibitors on the expression of intercellular adhesion molecule-1 (ICAM-1) in melanoma cells. METHODS: ICAM-1 protein of human melanoma cell a375 was detected by enzyme-linked immunosorbent, flow cytometry and Western blot analysis. Level of ICAM-1 mRNA in a375 was evaluated by Northern blot analysis. Adhesion of a375 to endothelial cell EC304 was analyzed by isotopic tracing. RESULTS: 5-Lipoxygenase inhibitors nordihydroguaiaretic acid, AA861 and MK886, could suppress the expression of ICAM 1 protein as well as of its mRNA in a375 cells and reduce the adhesion of a375 to EC304. CONCLUSION: 5-Lipoxygenase inhibitors can inhibit the expression of ICAM-1 in human melanoma cells and may be valuable for treatment of melanoma metastasis. PMID- 15132837 TI - Tanshinone prevents cancellous bone loss induced by ovariectomy in rats. AB - AIM: To investigate the skeletal effects of total tanshinone in ovariectomized rats by analyzing cancellous bone histomorphometry of fourth lumbar vertebrae (LV4) and proximal tibial metaphyses (PTM). METHODS: Four-month-old Sprague Dawley female rats were sham-operated and treated with vehicle or ovariectomized and treated with either vehicle, total tanshinone (200 mg x kg(-1) x d(-1), equivalent to 35 microg x kg(-1) x d(-1) of tanshinone II A and 16 microg x kg( 1) x d(-1) of cryptotanshinone), or 17alpha-ethynylestradiol (30 microg x kg(-1) x d(-1) as positive treatment group) starting one day post-surgery for 10 weeks. Double in vivo fluorochrome labeling was administered to all rats. The undecalcified longitudinal LV4 and PTM sections were cut and stained with Goldner's Trichrome (4-microm thickness) or unstained (8-microm thickness) for the bone histomorphometric analysis. RESULTS: A significant decrease in trabecular bone volume (BV/TV) and trabecular number (Tb.N) and a significant increase in osteoclast surface (OCS/BS) and mineralizing surface (MS/BS) were found in both LV and PTM of vehicle-treated OVX rats compared with sham controls. Tanshinone completely prevented the decreases in BV/TV and Tb.N and the increase in OCS/BS in the LV4, and partially prevented the decreases in BV/TV and Tb.N in the PTM of OVX rats. In addition, tanshinone increased trabecular thickness (Tb.Th) whereas it did not alter MS/BS. Moreover, tanshinone had no effect on uterine weight and body weight of OVX rats. Estrogen treatment increased BV/TV and Tb.N and decreased OCS/BS, but, also markedly decreased MS/BS and increased uterine weight in OVX rats. CONCLUSION: The current study demonstrated that the adequate supply of tanshinone prevented OVX-induced cancellous bone loss in rats through inhibition of elevated bone resorption. PMID- 15132838 TI - Expression of PDCD5, a novel apoptosis related protein, in human osteoarthritic cartilage. AB - AIM: To investigate the expression features of PDCD5 (programmed cell death 5 protein) in osteoarthritic and normal human cartilage, and speculate on its potential functions in the pathogenesis of osteoarthritis (OA). METHODS: Articular cartilage specimens were obtained from 30 patients with OA and 16 healthy patients at the time of arthroplasty. Expression of PDCD5 was detected by flow cytometry, immunofluorescence, RT-PCR and immunohistochemical analysis. RESULTS: Enhanced expression and nuclear accumulation of PDCD5 in OA chondrocytes were found. PDCD5-positive chondrocytes were mainly distributed in the superficial and deep zones of OA tissue sections, as opposed to, in the superficial and middle regions of normal healthy tissue sections. CONCLUSION: Since apoptotic chondrocyte death occurs more frequently in OA cartilage than in normal healthy cartilage and PDCD5 is an apoptosis-related protein, the different expression patterns of PDCD5 in OA cartilage from that in normal healthy cartilage indicate that PDCD5 is involved in the pathogenesis of OA. PMID- 15132840 TI - CYP2E1 mediated isoniazid-induced hepatotoxicity in rats. AB - AIM: To investigate the role of CYP2E1 in isoniazid (INH)-induced hepatotoxicity and the influence of rifampicin (RFP) on INH-induced liver injury. METHODS: Rats were treated with INH alone (100 mg/kg, ip) or co-administered with RFP (100 mg/kg, ig) for 10 d and 21 d. Hepatotoxicity was assayed by plasma enzymes (sALT, sAST) and histopathological examinations. Hepatic CYP2E1 activity was measured by aniline hydroxylase (ANH), and CYP2E1 mRNA expression was determined by RT-PCR. Plasma hydrazine concentration was determined by RP-HPLC. RESULTS: For a 10 d INH treatment, hepatic CYP2E1 level was increased to 3.7-fold over the control; liver impairment appeared after 21 d treatment, while CYP2E1 and plasma hydrazine were, respectively, increased to 4.6-fold and 1.7-fold. However, in INH-RFP group for 10 d, CYP2E1 and plasma hydrazine were, respectively, decreased by 13 % and 18 % over INH group; similarly, hepatic injury is equal to INH group appeared after 21 d, and CYP2E1 was further decreased by 26 %. Correlation analysis showed that sALT had a positive correlation with plasma hydrazine and with CYP2E1 activity; CYP2E1 activity was also markedly correlated with plasma hydrazine. And compared with control, there is no difference in changes of CYP2E1 mRNA expression in INH and INH-RFP treatment for 21 d. CONCLUSION: The metabolite of INH, hydrazine, plays an important role in INH-induced hepatotoxicity in rats. The induction of CYP2E1 by hydrazine is involved in the hepatotoxicity of INH. RFP does not exacerbate INH-induced hepatotoxicity in short term, which relates to down regulation of CYP2E1. PMID- 15132839 TI - Oridonin induces apoptosis of HeLa cells via altering expression of Bcl-2/Bax and activating caspase-3/ICAD pathway. AB - AIM: To study the mechanisms by which oridonin inhibited HeLa cell growth in vitro. METHODS: Viability of oridonin-induced HeLa cells was measured by MTT assay. Apoptotic cells with condensed nuclei were visualized by phase contrast microscopy. Nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis. Caspase activity was assayed using fluorometric protease assay. ICAD, Bcl-2, and Bax proteins expression were detected by Western blot analysis. RESULTS: Oridonin induced oligonucleosomal fragmentation of DNA and increased caspase-3 activity, on the other hand, reduced the expression of inhibitor of caspase-3-activated DNase (ICAD), a caspase-3 substrate, at 12 h in HeLa cells. Oridonin-induced DNA fragmentation, caspase-3 activation and down-regulation of ICAD expression were effectively inhibited by a caspase-3 inhibitor, z-DEVD-fmk (z-Asp-Glu-Val-Asp-fmk). However, pretreatment with an inhibitor of poly (ADP ribose) polymerase (PARP), 3, 4-dihydro-5-[4-(1-piperidinyl)butoxy]-1(2H) isoquinolinone (DPQ), did not suppress oridonin-induced HeLa cell death. In addition, oridonin-induced apoptosis was associated with an increase in the expression of the apoptosis inducer Bax, and a significant reduction in expression of the apoptosis suppressor Bcl-2 in mitochondria. CONCLUSION: Oridonin induces HeLa cells apoptosis by altering balance of Bcl-2 and Bax protein expression and activation of caspase-3/ICAD pathway. PMID- 15132841 TI - [Pathological diagnosis of interstitial lung disease]. PMID- 15132842 TI - [Clinicopathologic features of usual interstitial pneumonia and its differential diagnosis from idiopathic nonspecific interstitial pneumonia]. AB - OBJECTIVE: To investigate the clinicopathologic features of usual interstitial pneumonia (UIP) and its differential diagnosis from idiopathic nonspecific interstitial pneumonia (INSIP). METHODS: The clinical and pathological features of 15 UIP and 11 cases of INSIP, having received open or video-assisted thoracoscopic lung biopsies and having follow-up information were reviewed. RESULTS: UIP occurred more often in males over 50 years of age. Clinical findings included progressive shortness of breath, cough, sputum and crackles over both lung fields. High resolution computerized tomography (HRCT) showed patchy attenuation, especially over both lower lobes. Honeycombing was found in 8 cases. Histologically, UIP was characterized by scattered fibrotic foci, fibrosis (often associated with honeycombing) and pulmonary architectural destruction. It had a heterogeneous appearance, with alternating areas of normal lung, interstitial inflammation, fibrosis and honeycomb changes. The frequencies of fibroblastic foci, muscle sclerosis, honeycomb changes, diffuse fibrosis and pulmonary architectural destruction in UIP and INSIP were 100% and 27.3% (P<0.001), 80.0% and 36.4% (P<0.05), 86.7% and 27.3% (P<0.001), 100% and 54.5% (P<0.01) and 100% and 45.5% (P<0.05), respectively. The response rate to glucocorticoid was 26.7% and 72.7% (P<0.05) in UIP and INSIP respectively. CONCLUSIONS: The distinction between UIP and INSIP is difficult if based on clinical examination alone. HRCT is helpful for differential diagnosis in some difficult cases. Definite diagnosis depends mainly on open lung biopsies. Key histologic features of UIP include heterogeneous appearance with interstitial inflammation, fibroblastic foci, scar formation and honeycomb changes. PMID- 15132843 TI - [Histologic features and pathologic diagnosis in usual interstitial pneumonia]. AB - OBJECTIVE: To study the pathologic features, differential diagnosis and role of open lung biopsies (OLB) in usual interstitial pneumonia (UIP). METHOD: The authors reviewed the pathologic, clinical and radiologic features of five cases of UIP (one autopsy case and four OLB cases), with follow-up information. RESULTS: The typical histologic features were a non-uniform distribution of alveolar inflammation, fibroblastic foci, interstitial fibrosis and honeycomb change. There also was associated metaplasia of bronchiolar epithelium, type II pneumocyte hyperplasia and accumulation of alveolar macrophages. CONCLUSIONS: Characteristically, UIP exhibits temporal heterogeneity under low-power light microscopy, which includes changes in both the early and end stages. Open lung biopsy is an important diagnostic adjunct for suitable patients with atypical radiologic features on computerized tomography. Correlation between clinical, radiologic and pathologic findings is also essential for a correct diagnosis. PMID- 15132844 TI - [Pathologic diagnosis and differential diagnosis of pulmonary Langerhans' cell histiocytosis]. AB - OBJECTIVE: Morphologic findings of pulmonary Langerhans' cell histiocytosis were analyzed in order to delineate diagnostic features. METHODS: H&E staining and immunohistochemical studies were performed on 7 cases of pulmonary Langerhans' cell histiocytosis. RESULTS: Infiltration by Langerhan's cells was obvious in all 7 cases. Inflammatory cell infiltrates, interstitial fibrosis and focal necrosis may also be seen. The cells expressed S-100 (7/7), CD68 (3/7), and CD1a (5/5). CONCLUSIONS: In case there is radiologic suspicion of Langerhans' cell histiocytosis, pulmonary biopsy is strongly advised for a definitive diagnosis. S 100 and CD1a immunostaining is also helpful in this respect. PMID- 15132845 TI - [Clinicopathologic analysis of organizing pneumonia in elderly autopsies]. AB - OBJECTIVE: To study the clinicopathologic characteristics of organizing pneumonia of the autopsies in elder and to analyze the possible underlying etiologic factors. METHODS: Ninety-five cases of organizing pneumonia were found from 635 elderly autopsy reports of the Beijing Hospital since 1980. The morphologic and imaginal features were analyzed. RESULTS: The foci of organizing pneumonia in our series were often small, patchy, scattered and adjacent to other lung lesion. There were, however, some variations in cases with different underlying conditions. The conditions associated with organizing pneumonia, as detected in our series, were: (1) 36 cases with suppurative inflammation, lung abscesses and chronic relapsing pneumonia; (2) 17 cases with fungal or viral infection; (3) 16 cases with aspiration pneumonia; (4) 5 cases with radiation pneumonia. We observed that it was not uncommon for organizing pneumonia coexisting with atelectasis and chronic pleuritis. The image of organizing pneumonia was varied. CONCLUSIONS: Organizing pneumonia is a common finding in autopsies of the elder. It occurs in association with many diseases and the basic pathologic changes are similar. All of which represent secondary phenomenon. Possible etiologic factors include infection (due to bacteria, fungi or virus), aspiration and radiation. The possibility of organizing pneumonia should be considered if the shadow of lung is undisappeared in imagin. PMID- 15132846 TI - [Clinicopathologic analysis of paragonimiasis]. AB - OBJECTIVE: To study the clinical manifestations and pathologic findings of paragonimiasis. METHODS: Nine cases of paragonimiasis diagnosed in the Peking Union Medical College Hospital during the past 20 years were studied, with literature review and analysis of the epidemiological, clinical and pathologic characteristics. RESULTS: Of the 9 cases studied, 4 came from the northeast China and 5 from Beijing. Eight cases had a history of eating raw crabs. Most had symptoms including fever, chest discomfort or pain, and hemoptysis or rusty sputum. All had the following common pathologic features: formation of irregular lacunae or sinus tracts, Charcot-Leyden crystals, sometimes paragonimus body parts and/or eggs, and eosinophil infiltration in the adjacent tissues. CONCLUSIONS: Paragonimiasis is not as uncommon as previously thought. The incidence is increasing in some cities due to movement of populations. The pathological diagnosis can be confirmed by finding paragonimus body parts and/or eggs. Diagnosis can also be made by correlation with other typical pathologic features, clinical history, immunologic findings and radiography. Paragonimiasis needs to be differentiated from pulmonary tuberculosis and cancer. PMID- 15132847 TI - [Pathologic diagnosis and subtyping of lymphoma in bone marrow biopsies using histologic examination, immunohistochemistry and gene rearrangement studies]. AB - OBJECTIVE: To assess the value of histologic examination, immunohistochemistry and gene rearrangement studies in the diagnosis and subtyping of lymphoma with bone marrow involvement (BMI). METHODS: Sixty-two formalin fixed, paraffin embedded bone marrow biopsy specimens were studied. Immunohistochemical and immunoglobulin heavy chain (IgH) and T-cell receptor gene rearrangement studies were performed in each case. RESULTS: Chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) demonstrated mainly and interstitial infiltration by dysplastic lymphocytes, with intertrabecular nodular arrangement or in dispersion. Sometimes, pseudofollicles may be noted. A predominantly para- or intertrabecular infiltration by nodules of lymphoma cells was characteristic of follicle center cell lymphoma (FCL) cases. In most lymphoplasmacytoid lymphoma (LPL) cases, there was infiltration by small lymphocytes and plasma cells between bony trabeculae. In marginal zone cell lymphoma (MZL), vague inter- or para trabecular nodules of polymorphic lymphoma cells with clear cytoplasm might be noted. Small to medium-sized dysplastic lymphocytes, with absence of paraimmunoblasts or pseudofollicles, were the most frequent findings in mantle cell lymphoma (MCL). Hairy cell leukemia (HCL) might be identified by the presence of distinct cell membrane and abundant clear cytoplasm, resulting in a "fried-egg" appearance. Tumor cells with large nuclei and eosinophilic nucleoli were characteristically seen in lymphomatosis diffusa (Hodgkin's disease, HD). In T-cell non-Hodgkin lymphoma with BMI, dispersed or clusters of intertrabecular neoplastic lymphoid cells with clear cytoplasm and gyriform nuclei were often observed. In diffuse large B-cell lymphoma (DLBL), the tumor cells were large and isolated or arranged in diffuse pattern. Immunohistochemically, a panel of markers, including CD3 CD20, and CD79 are valuable for the differential diagnosis of T- and B-cell lymphomas. The neoplastic cells in MCL were cyclin D1- and CD5 positive, while BCL2- and CD10-positivity was characteristic for FCL. CLL/SLL cells might be stained with CD5 and CD23, in addition to CD20 and CD79. CD25 expression might be noted in HCL: the positivity for CD15, CD30 and fascin suggests HD. There was a higher positivity rate for IgH gene rearrangement in CLL/SLL, LPL MZL and DLBL (80%, 60%, 66.7%, 70% respectively) and for T- cell receptor gamma gene rearrangement in T-cell lymphoma (66.7%). CONCLUSION: A combination of histopathology, immunohistochemistry and IgH / T-cell receptor gamma gene rearrangement studies may be of aid to the diagnosis and subtyping of lymphoma with BMI, especially if there is only a small number of tumor cells present in the specimen. PMID- 15132848 TI - [Glial abnormalities in progressive supranuclear palsy and corticobasal degeneration]. AB - OBJECTIVE: To study pathologic features of glial cells in progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) and to explore their pathologic significance. METHODS: Brain tissues from 2 cases with PSP and 3 cases with CBD, all confirmed by autopsies, were examined by routine neuropathologic methods, Gallyas-Braak staining and tau immunostaining. Brain tissues from 6 Alzheimer's disease cases, 4 cases with Parkinson's disease and 6 elderly with no neurologic abnormality were used as controls. RESULTS: Gallyas-Braak staining demonstrated tuft-shaped astrocytes and coiled-body oligodendroglial cells in the brain tissues of 2 cases with PSP and 3 cases with CBD. The tuft-shaped astrocytes appeared prominently in the frontal and parietal cortex, basal ganglia and grey matter of the brainstem. The coiled-body oligodendroglial cells were distributed widely in the white matter of the frontal and parietal lobes, basal ganglia, brainstem and cerebellum. However, astrocytic plaques, composed of degenerative stubby processes with radiating arrangement, only appeared in the frontal, parietal and cingular cortex, as well as in the striatum of 3 cases with CBD. The astrocytic plaques and tuft-shaped astrocytes coexisted in the same areas, including parietal and cingular cortex and striatum, in CBD. All these glial abnormalities showed tau-positive immunoreaction not found in control cases. CONCLUSIONS: The tuft-shaped astrocytes and coiled-body oligodendroglial cells are common glial morphologic features of both PSP and CBD. Astrocytic plaques are also characteristically seen in CBD. PMID- 15132849 TI - [Expression of proteins in p53 (p14ARF-mdm2-p53-p21WAF/CIP1) pathway and their significance in exocrine pancreatic carcinoma]. AB - OBJECTIVE: To investigate the expression of p14(ARF), p53, mdm2 and p21(WAF/CIP1) proteins and their relationship in exocrine pancreatic carcinoma. METHODS: Specimens of pancreatic carcinoma, adjacent non-neoplastic pancreatic tissue and pancreatic benign lesions were examined for p14(ARF), p53, mdm2 and p21(WAF/CIP1) protein expression by tissue microarray technique and immunohistochemistry. RESULTS: The expression of p14(ARF), p53, mdm2 and p21(WAF/CIP1) proteins in pancreatic carcinoma were 35.3% (59/167), 57.5% (96/101), 64.1% (107/167) and 39.5% (66/167) respectively. The expression of p53 proteins was increased in pancreatic carcinoma (P < 0.01), while the expression of p14(ARF) and p21(WAF/CIP1) proteins was reduced (P < 0.05), as compared with that in non neoplastic pancreatic tissue. p21(WAF/CIP1) protein expression in pancreatic carcinoma significantly correlated with the age of patients and perineural invasion (P < 0.05). p53 protein expression correlated significantly with tumor differentiation, lymph node metastasis and perineural invasion (P < 0.05). Mdm2 protein expression correlated significantly with tumor differentiation (P < 0.05), while p14(ARF) protein expression correlated significantly with the age of patients and metastasis (P < 0.05). There was also statistic correlation between the expression of these four genes (P < 0.05). CONCLUSIONS: Overexpression of p53 and mdm2 and loss of p14(ARF) and p21(WAF/CIP1) expression may contribute to the pathogenesis of pancreatic carcinoma. These proteins play a critical role in cell cycle arrest and apoptosis after DNA damage through p14(ARF)-mdm2-p53 p21(WAF/CIP1) pathway. Detection of p53 and Mdm2 protein overexpression may be useful in evaluation of the aggressiveness of pancreatic carcinoma. PMID- 15132850 TI - [Pathologic diagnoses of core needle biopsies of the mediastinum]. AB - OBJECTIVES: To assess the diagnostic accuracy and to study the histologic typing of mediastinal lesions using core needle biopsies. METHODS: The histopathology and immunophenotype of 65 mediastinal core needle biopsy specimens were studied retrospectively by light microscopy and immunohistochemical staining (ABC method). Gene rearrangement studies were performed in some of the non-Hodgkin's lymphomas cases using PCR. Follow-up records were also analyzed. RESULTS: Morphologically, all specimens showed a combination of epithelioid cells, lymphoid cells and fibrous tissue in different proportions. The pathologic diagnoses included lymphoma (21 cases), pulmonary carcinoma (20 cases), thymoma (14 cases), thymic carcinoma (4 cases), seminoma (3 cases) and chronic inflammation (1 case). Definitive diagnosis was not possible in 2 cases due to insufficient material. The tumor cells in lymphoma (21 cases) expressed CD20, CD3, TDT, CD30, CD15 or EMA, depending on their histologic subtypes. Tumor cells in the 17 pulmonary carcinoma cases expressed cytokeratin (CK), except 3 cases of small cell carcinoma of lung. Synaptophysin, chromogranin A and neuron-specific enolase were all positive in the 10 cases of small cell carcinoma of lung and 1 case of thymic small cell carcinoma (which was also CD5 negative). The 3 cases of adenocarcinoma of lung showed positivity for thyroid transcription factor-1 (TTF 1) and they were negative for CD5. The 14 thymoma cases expressed CK, CD3 or CD20. The 3 thymic carcinoma cases expressed CK and CD5. Placental-like alkaline phosphatase (PLAP) was positive in 3 seminoma cases which were CK-negative. Immunoglobulin heavy chain gene was rearranged in the 3 cases of diffuse large B cell lymphoma and 1 B-cell anaplastic large cell lymphoma case. T-cell receptor beta gene was rearranged in 5 T-cell lymphoblastic lymphoma cases. CONCLUSIONS: Microscopic assessment of tissue samples from mediastinal core needle biopsies should be made in combination with clinical and radiologic information. Ancillary investigations, including immunohistochemical staining and/or gene rearrangement studie, are needed in both non-lymphoma and lymphoma cases of mediastinum. PMID- 15132851 TI - [Value of special stains and immunohistochemistry in the diagnosis of renal epithelial neoplasms]. AB - OBJECTIVE: To study the diagnosis and differential diagnosis of renal epithelial neoplasms. METHODS: Ninety-one cases of renal epithelial neoplasms with detailed pathologic records were enrolled. In addition to microscopic examination, Mowy's colloidal iron staining and immunohistochemical studies (CD10, vimentin and CK7) were also performed. RESULTS: Among the 91 cases, there were 78 (86%) clear cell renal carcinoma cases, 8 (9%) papillary renal carcinoma cases, 4 (4%) chromophobe renal carcinoma cases and 1 (1%) renal oncocytoma case. Sixty-three of the 78 clear cell renal carcinoma cases were positive for CD10 and 69 were positive for vimentin (81% and 88% respectively), with prominent cell membrane staining. The majority (74/78) of clear cell renal carcinoma were negative for CK7. All 17 clear cell renal carcinoma cases showed negative or focal coarse droplet-like staining pattern for Mowy's colloidal iron stain. All 4 chromophobe renal cell carcinoma cases showed prominent cell membrane staining for CK7 and blue reticular staining pattern for Mowy's colloidal iron stain. All of which were negative for CD10 and vimentin. The case of renal oncocytoma failed to react with antibodies to CD10, vimentin and CK7, or Mowy's colloidal iron stain. CONCLUSIONS: CD10, vimentin, CK7 and Mowy's colloidal iron stains have proved to be useful in differential diagnosis of common renal tumors which may not be easily distinguished on the basis of histologic examination alone. PMID- 15132852 TI - [Study of tracheal regeneration after injury induced by 5-fluorouracil in rats]. AB - OBJECTIVE: Localization of tracheal stem cells in rat trachea. METHODS: Extracorporeal tracheal injury (Wistar rats) was induced by 5-FU. The process of regeneration was observed and analyzed by light microscopy, electron microscopy, and immunohistochemistry. RESULTS: Twelve hours after treatment with 5-FU, the tracheal epithelium shed and cells with naked nuclei were seen located sparsely on the basement membrane. Six hours after removal of 5-FU, the tracheal rings were covered with flattened epithelium. These cells were poorly differentiated under electron microscopy. Immunohistochemistry showed few proliferating cell nuclear antigen (PCNA)-negative cells sparsely scattered among PCNA-positive cells on the basement membrane. Nine hours later, electron microscopy found that these cells differentiated into mucous cells and ciliated cells. Forty-eight hours later, the tracheal rings were entirely covered by pseudostratified ciliated columnar epithelium. CONCLUSIONS: A small number of G(0) cells with naked nuclei are located sparsely on the basement membrane of the trachea. Tracheal epithelium regenerates by proliferation and differentiation of these cells. It is likely that some of these G(0) cells on the tracheal basement membrane represent tracheal stem cells. PMID- 15132853 TI - [Effects of P2Y receptor activation on prostatic cancer cells requiring ERK1/2 or p38 cascade]. AB - OBJECTIVE: To determine the role of extracellular signal-regulated kinase (ERK1/2) and p38 cascades in P2Y receptor-evoked effects on prostatic cancer cells. METHODS: Highly metastatic prostatic cancer cells 1E8 were transfected with dominant-negative MAPK kinase 1 (KA-MEK1). The activation of ERK1/2 was determined by Western blot technique. The role of ERK1/2 and p38 cascades in P2Y receptor-evoked effects on in vitro growth, colony formation and in vitro invasion was detected by cell count, soft agar colony formation assay and in vitro invasion assay. The effect of ATP on apoptosis was detected by flow cytometry. RESULTS: ERK1/2 activity in 1E8-KA-MEK1 transfectants was significantly suppressed by dominant-negative MEK1 transfection. After culture of 6 days, 1E8-KA-MEK1 transfectants exhibited a growth inhibition of 71% as compared with 1E8-pcDNA3 control. Moreover, after continuous treatment with 100 micro mol/L ATP for 6 days, the growth of 1E8-KA-MEK1 transfectants was further inhibited by an additional 17.2%. Pretreatment with 10 micro mol/L p38 inhibitor SB203580 antagonized the effect of ATP-induced additional growth inhibition, suggesting that ERK1/2 and p38 pathways play an important role in ATP-induced growth inhibition. In soft agar assay, 1E8-KA-MEK1 transfectants formed smaller colonies and exhibited a 75% decrease in colony formation (as compared with control). Further treatment with ATP or SB203580 plus ATP did not show significant effect on colony formation of 1E8-KA-MEK1 cells, implying a potential role of ERK1/2, instead of p38, in P2Y receptor-mediated inhibitory effect on colony formation. In in vitro invasion assay, 1E8-KA-MEK1 cells showed a 41% decrease in passing through matrigel-coated membranes, as compared with control. Treatment with ATP could restore their invasive ability, and this effect by ATP could be blocked by pretreatment with SB203580, indicating the involvement of both ERK1/2 and p38 pathways in invasive ability of prostatic cancer cells. CONCLUSIONS: The effects of ATP on in vitro growth, invasion and colony formation of prostatic cancer cells depend on the status of P2Y receptor activation by different treatment protocols. Continuous activation of P2Y receptor results in growth inhibition and transient activation of P2Y receptor stimulates in vitro invasion of prostatic cancer cells. Both ERK1/2 and p38 pathways are responsible for these effects; but only the ERK1/2 pathway is involved in regulation of colony formation of prostatic cancer cells. PMID- 15132854 TI - [Relationship between the expression of Fas, bcl-2 and apoptosis of cardiomyocytes in experimental myocardial ischemia in rats]. AB - OBJECTIVE: To explore the relationship between expression of Fas, bcl-2 and apoptosis of cardiomyocytes in myocardial ischemia in rats. METHODS: Myocardial ischemia was experimentally induced by ligating the left coronary artery. The rate were killed from 10 minutes to 7 days after the operation. Apoptotic myocardial cells were detected by TUNEL method. The expression of Fas and bcl-2 was studied by ABC immunohistochemistry. RESULTS: Cardiomyocytic apoptosis appeared from 3 to 36 hours after ischemia. This phenomenon however could not be detected by TUNEL method 7 days after ischemia. The expression of Fas could be detected by ABC immunohistochemistry from 3 hours to 7 days after ischemia, and the expression of bcl-2 from 10 minutes to 7 days. Cardiomyocytic apoptosis and Fas / bcl-2 expression appeared in different regions of myocardium: apoptosis in the ischemic regions, while Fas and bcl-2 expression in regions without obvious ischemia. CONCLUSION: In rats, Fas and bcl-2 may not directly regulate apoptosis of cardiomyocytes in case of myocardial ischemia. PMID- 15132855 TI - [Sulfur dioxide-induced liver pathology]. AB - OBJECTIVE: To investigate effects of short-term sulfur dioxide inhalation to the liver. METHODS: Haematoxylin and eosin staining (HE) and transmission electron microscopy (TEM) were used to study the pathologic changes in mice liver after sulfur dioxide (SO(2)) inhalation. RESULTS: Exposure to 56 mg/m(3), 112 mg/m(3) 168 mg/m(3) SO(2) caused increasingly severe liver injuries, as detected by HE staining and TEM. The morphologic changes included spotty necrosis with lymphocyte, monocyte, and neutrophil infiltration, fatty degeneration of hepatocytes with dilatation of rough endoplasmic reticulum and dissociation of ribosomes, as well as degeneration of mitochondria and karyorrhexis. CONCLUSION: SO(2) inhalation can cause marked liver injury in experimental settings. PMID- 15132856 TI - [Idiopathic pulmonary fibrosis/usual interstital pneumonia]. PMID- 15132857 TI - [Nature and histogenesis of pulmonary sclerosing hemangioma]. PMID- 15132858 TI - [Body mass index reference norm for screening overweight and obesity in Chinese children and adolescents]. AB - OBJECTIVE: To establish a national body mass index (BMI) reference norm for the purpose of screening and more active prevention and cure on overweight and obesity in Chinese children and adolescents. METHODS: The 2000 Chinese National Survey on Students Constitution and Health was used as reference population. In total there were more than 244.2 thousands of primary and secondary Han nationality students aged 7 through 18 years old included in this study. The BMI distribution of various Chinese children and adolescent groups were compared with the NCHS international norm, the percentage values and with advanced ages, were calculated. In the beginning, three temporary norms were set up, using the different combination of P(85), P(90) and P(95) BMI percentages. Based on the intersect testing and varifying of physiological, lipidemia biochemical and body composition measures, the best norm was then selected. B-spline smoothing method was used to correct the curves, both for males and females, composed by cut-off points at different ages. RESULTS: Using samples from the costal developed metropolis, the BMI curves successfully overcame the shortcomings of low and depressive phenomenon of the total population, in particular after the mid adolescent period. The temporary Norm II, composed by cut-off points of P(85) for overweight and P(95) for obesity, was found to be the best among the three temporary norms, both shown by its sensitivity and specificity. 24 and 28 were used as cut-off points for overweight and obesity in this norm, both for males and females aged 18 years. These two cut-off points were consistent with those shown in the Body Mass Index Reference Norm for Screening Overweight and Obesity in Chinese Adults. Three samples from Beijing, Henan and Sichuan, used as representatives of upper, middle and relative low levels of physical growth of children and adolescents in China, were used for extra tests. The screening results showed that among the male and female subjects aged 7 through 18 years, the prevalence rates were 17.00% and 9.46% for overweight, 9.99% and 6.47% for obesity in Beijing while 10.86% and 6.64% for overweight, 4.27% and 3.07% for obesity in Henan, and 6.95% and 4.23% for overweight and 2.84% and 2.09% for obesity in Sichuan, respectively. Data clearly showed that the results were in accordance with the actural situation seen in the Chinese students in 2000. CONCLUSION: The newly established reference norm seemed to be good for prospective studies as it considered the facts regarding the rapid and strong tendency of secular growth changes seen in the Chinese children and adolescents. It also decreased the difference of growth levels between the Chinese children and adolescents and those of the international norms. This norm was particularly developed for the Chinese which was also in consistent with the Eastern Asia ethnic characteristics so could be promoted in China. PMID- 15132859 TI - [Dynamic analysis on the prevalence of obesity and overweight school-age children and adolescents in recent 15 years in China]. AB - OBJECTIVE: In order to develop strategies on prevention and cure for obesity, a study regarding dynamic analyses on the prevalence of obesity and overweight of Chinese children and adolescents was carried out. METHODS: Data of more than 980 thousands students of Han nationality, aged 7 through 18 years old were collected from National Surveys on Chinese Students' Constitution and Health which were carried out in 1985, 1991, 1995 and 2000. Samples were divided into four groups as "metropolis", "medium and small sized cities", "prosperous village" and "below averaged village". Study targets were screened using the new criterion-the "Reference Norm for Screening Overweight and Obesity in Chinese Children and Adolescents" which was set up by the Working Group on Obesity in China. The prevalence of overweight and obesity in various groups in different years were compared and analyzed. RESULTS: The prevalence rates of obesity in 1985 including from the metropolis area, were only 0.2% and 0.1% for boys and girls, and the prevalence of overweight was between 1% and 2% indicating that was no trend of obesity epidemic then. However, a rapid increase of overweight prevalence has been noticed since the early 1990s, and the increments were more seen in the urban than in the rural areas and more in boys than in girls. In the most developed cities including Beijing, the prevalence rates of obesity appeared to be 4.7%, 3.8% and 3.2%, among three groups with higher risk: the 7 - 9, 10 - 12 year-old boys and 7 - 9 year-old girls, respectively. Both high prevalence rates of malnutrition and overweight were also found in that period. Around 1995, a large scale of increments of overweight were found both in the urban and rural groups. In the most developed metropolis, the prevalence of overweight was two to three folds more than that of 10 years ago. The prevalence of obesity were 6% - 8% for boys and 4% - 6% for girls, respectively. Since 2000, most of the Chinese metropolis have started the so called 'overall increment period' of obesity. The prevalence rates of obesity plus overweight had reached 25.4%, 25.5%, 17.0% and 14.3% for boys aged 7 - 9 years and 10 - 12 years, and girls aged 7 - 9 years and 10 - 12 years, respectively. Among them, the prevalence rates of obesity were 12.9% and 9.1% for boys aged 7 - 9 and 10 - 12 years, which had already reached the average level seen in the medium-developed countries in the world. Although the increments of overweight were high, the prevalence of obesity was still low in most of the other groups: 4.8% for boys and 2.6% for girls in the "medium sized" group, 1.5% for boys and 1.7% for girls in the "prosperous village" group, and 0.9% for boys and 1.2% for girls in the "below-averaged village" group, respectively. CONCLUSIONS: The prevalence of obesity in Chinese children and adolescents was considered to be still relatively low. However, the rapid increasing of both obesity and overweight, in both urban and rural areas would arouse special attention. Comprehensive strategies of intervention should include periodical monitoring, education on pattern of nutrition, oxygen-consuming physical exercises and healthy dietary behavior. Two areas on obesity prevention to protect the child from becoming overweight in early ages, and to control the tendency of obesity from overweight, should be emphasized in order to improve the health status and life quality of the Chinese children and adolescents. PMID- 15132860 TI - [The study on relationship of body mass index and blood pressure in children and adolescents of Beijing]. AB - OBJECTIVE: To identify the relationship of body mass index (BMI) and blood pressure in 7 - 15 years children and adolescents of Beijing so as to provide scientific basis for early prevention of hypertension and to provide evidence for verification on the category criterion of overweight and obesity in children and adolescents of China, recommended by the Working Group on Obesity in China (WGOC) to sensitively distinguish the blood pressure in normal weight, overweight and obesity populations. METHODS: A cross-sectional survey on epidemiological characteristics of obesity with stratified cluster sampling method carried out in Beijing in April and May, 2000. 5155 students aged 6 - 15 years were selecte das research subjects. The category criterion of overweight and obesity in children and adolescents of China was recommended by WGOC, the diagnostic criterion of hypertension in children was recommended by CDC in the USA. Statistics analysis system (SAS 8.1) including partial person correlation analysis, t-test, chi(2) test and logistic multi-factors regression analysis was used to analyses the data from 4982 subjects aged 7 - 15 years. RESULTS: (1) after the age and gender were adjusted, the BMI positive correlation with systolic blood pressure (SBP) and diastolic blood pressure (DBP) was found independent in 7 - 15 years children and adolescents (P < 0.0001) and the partial relation coefficients(r) between BMI and SBP and DBP were 0.323 87 and 0.245 88 respectively. (2) the means of SBP and DBP in obesity group were significantly higher then overweight, while overweight was significantly higher then normal weight group (P < 0.0001). (3) the prevalence rates of hyper-SBP, hyper-DBP and hypertension were significantly different (P < 0.0001). When compared with the normal weight group, the relation risk (RR) for hypertension in overweight group and obesity group were 2.96 and 4.85 respectively. The prevalence rates of hypertension in overweight and obesity group were 19.70% and 24.22% respectively. (4) the results of logistic multi factors regression analysis showed that both age and weight were effecting on hyper-SBP, hyper-DBP and hypertension (P < 0.0001). After age was adjusted, the RR for hypertension was 2.62, and their confidence interval (CI) was 2.36 - 2.91 in obesity or overweight, between overweight and normal weight. CONCLUSION: (1) the BMI positive correlation with SBP and DBP was found independent in 7 - 15 years children and adolescents of Beijing, and the risk for hypertension maybe increased when these people with overweight and obesity, it is very important for hypertension prevention and control that overweight and obesity prevention and control in children and adolescents. (2) the sensitivity of the category criterion of overweight and obesity in children and adolescents of China, recommended by WGOC have been verified on distinguish the blood pressure in normal weight, overweight and obesity populations. PMID- 15132861 TI - [The relation between body mass index and percentage body fat among Chinese adolescent living in urban Beijing]. AB - OBJECTIVE: To study the relation between body mass index (BMI) and percentage body fat (PBF) in Chinese adolescent, and to compare the prevalence of overweight and obesity using BMI or PBF standards. METHODS: BMI from 757 girls with an average aged of 10.1 years in the rural areas, 165 girls average aged 13.5 years in suburbs and 172 boys average aged 13.7 years in suburb of Beijing, were measured. Their body compositions were also measured by dual-energy X-ray absorptiometry (DEXA). RESULTS: BMI was found closely correlated with PBF in each age group of rural and suburb girls and suburb boys with the correlation coefficient(r) of 0.59 - 0.83. When age, height and pubertal development were controlled, r became 0.54 - 0.88. The prevalence rates of obesity in rural girls, suburb girls and suburb boys were 33.1%, 21.8% and 21.5%, when PBF cutoff values (girls: PBF >or= 35%, boys: PBF >or= 25%) were used. However, the rates became 2.4%, 3.0% and 4.0% when BMI cutoff values of International Obesity Task Force (IOTF) were used. Compared with PBF cutoff values for obesity, the IOTF recommended BMI cutoff values had relatively lower sensitivity (7.3% - 18.9%) and higher specificity (100%). CONCLUSION: BMI correlated well with PBF in Beijing adolescent. IOTF-BMI cutoff values showed low sensitivity and high specificity to Chinese adolescent which might be suitable for identifying obesity but not for the purpose of screening. PMID- 15132862 TI - [Validation of lipids on body mass index reference recommended by Obesity Working Group, International Life Science Association of China]. AB - OBJECTIVE: To assess the relationship between overweight, obesity and blood lipid profiles of children and adolescents and to validate body mass index (BMI) cutoff points for overweight and obesity screening to Chinese children and adolescents, recommended by Working Group of Obesity, China (WGOC), International Life Science Association. METHODS: 2293 children and adolescents (1124 males and 1169 females), aged between 10 and 18 years, were randomly selected as samples from 6 schools in Beijing area. Fasting serum lipids including total cholesterol (TC), total triglycerides (TG), high density lipoprotein cholesterol (HDLC), thropometrical index as weight and height were measured. BMI equals to weight in kilograms were then divided by the square of height in meters. RESULTS: According to BMI cutoff points recommended by WGOC, samples fell into 3 groups including normal group (BMI < 85 percentiles), overweight group (BMI >or= 85 and < 95 percentiles) and obesity group (BMI >or= 95 percentiles). Results clearly showed an increase of both serum TC and TG and a decrease of HDLC when BMI was increasing, among most age groups regardless of sex difference and the difference among BMI groups was statistically significant (P < 0.05 or P < 0.01). CONCLUSIONS: Results of this study indicated that there was an obvious dose effect relationship between BMI and lipid profiles which accounted for some rationality of the BMI cutoff points recommended by WGOC. The authors reckoned the findings important to managing relevant adult diseases during childhood, in China. PMID- 15132863 TI - [Study on the correlation between positive rates of SARS RNA in clinical confirmed SARS patients and the appearance of RNA in relation to the development of the disease]. AB - OBJECTIVES: To study the correlation between positive rates of RNA in clinical confirmed severe acute respiratory syndrome (SARS) patients and its appearance in relation to the development of the disease in order to provide scientific basis for early diagnosis, effective prevention and treatment of the disease. METHODS: One-step reverse transcription-polymerase chain reaction (RT-PCR) was used to amplify the SARS RNA in the clinical specimens from different courses of the disease. The representative amplicons were then sequenced. Chi-square for trend test was performed to study the correlation between positive rates of RT-PCR and at different periods after the onset of the disease. RESULTS: The fragments amplified from the sputum specimens of SARS patients were shown to share 100% homology with the published SARS-associated coronavirus. Of the different clinical specimens, positive rate in the stools appeared to be the highest (21.55%). Chi-square for trend test revealed that the positive rates of stools and sputa of SARS patients decreased with the development of the disease (chi(2) for trend = 12.55 and 16.408, P = 0.0004 and P = 0.000 05 respectively). CONCLUSION: One-step RT-PCR proved to be an effective method for the detection of SARS-associated coronavirus from clinical specimens. Data as indicated that the positive rates of SARS coronavirus were decreasing in SARS patients along with the disease progression. PMID- 15132864 TI - [Dynamics of peripheral blood lymphocytes and their subpopulations in patients with severe acute respiratory syndrome]. AB - OBJECTIVE: To study on the dynamics of peripheral blood lymphocytes and their subpopulations in patients with severe acute respiratory syndrome. METHODS: Using flow cytometry, the absolute numbers of peripheral blood lymphocytes and their subpopulations in 240 SARS patients (696 specimens) and 51 individuals as controls, were counted and compared. RESULTS: The absolute numbers of peripheral blood lymphocytes and their subpopulations (CD45, CD3, CD4, CD8) were 1298 +/- 785, 897 +/- 606, 510 +/- 372, 362 +/- 263/mm(3), respectively, significantly lower in SARS patients as compared to the normal controls (2024 +/- 423, 1391 +/- 289, 795 +/- 129, 551 +/- 183/mm(3)). Of SARS patients, severe group (1095 +/- 740, 740 +/- 562, 419 +/- 346, 304 +/- 244/mm(3)) had lower counts than that of mild group (1404 +/- 788, 991 +/- 612, 564 +/- 378, 396 +/- 267/mm(3)), and in group with deaths (587 +/- 493, 369 +/- 371, 204 +/- 191, 150 +/- 130/mm(3)) was lower than that of recovery group (1355 +/- 776, 948 +/- 603, 539 +/- 375, 382 +/ 263/mm(3)). There were significant differences (P < 0.01) for CD45, CD3, CD4, CD8, but with no significant difference (P > 0.05) for CD4/CD8 ratio between severe and mild, recovery and death groups. The lymphocytes and their subpopulations (CD45, CD3, CD4, CD8) declined in the 1st week and to the lowest level (977 +/- 579, 641 +/- 466, 360 +/- 275, 270 +/- 216/mm(3)) in the 2nd week. Then the lymphocytes and their subpopulations gradually increased during the recovery of the disease. CONCLUSION: The absolute numbers of peripheral blood lymphocytes and their subpopulations in SARS patients might be used as one of the methods for diagnosis on the severity and prognosis of the disease. PMID- 15132865 TI - [Early screening of cerebral palsy during infancy using 'Infant Motor Malfunction Profile' in the communities of two cities]. AB - OBJECTIVE: To evaluate the prospective validity and predictive value of cerebral palsy during infancy, using 'Infant Motor Malfunction Profile'. METHODS: Items of motor milestone in the profile was used as the first step to screen cerebral palsy on 8137 infants who were screened at 3, 6, 9, 12 months of age. The positive cases who had one of 7 developmental abnormal motor milestone items were examined using the items of primitive reflexes or postural reaction as the secondary step of cerebral palsy screening. The cases who had at least one abnormal item of primitive reflex or postural reaction received neurological examination by doctors to diagnose cerebral palsy. RESULTS: According to our data, sensitivity, specificity, positive prospective value, positive and negative likelihood ratio of the profile in screening cerebral during infancy were 94.1%, 99.8%, 48.5%, 449.6 and 0.06, respectively. CONCLUSION: The profile seemed to be an acceptable instrument for early identification of cerebral palsy. PMID- 15132866 TI - [Study on the prevalence of Tourette syndrome in children and juveniles aged 7-16 years in Wenzhou area]. AB - OBJECTIVE: To study the prevalence and distribution of Tourette syndrome (TS). METHODS: Nine thousand, seven hundred and forty-two children and juveniles aged 7 - 16 years in Wenzhou were studied, using cluster random sampling method. RESULTS: The prevalence of TS among school-age children was estimated to be 0.43% (0.74% for males and 0.07% for females). The prevalence of male children and juveniles was higher than that of female children and juveniles (chi(2) = 25.09, P < 0.001, prevalence ratio = 10.95, prevalence ratio 95% CI: 3.38 - 35.46). The highest prevalence of TS was between 9 - 10 years old. The mean age at onset of TS was 7.7 +/- 2.7 years, with 45.2% of them among 6 - 7 year olds. The rate of delayed diagnosis and rates of misdiagnosis and misclassification of the syndromes were 78.6%, 42.9% and 23.8%, respectively. CONCLUSION: Tourette syndrome had been a common disease of children and juveniles in Wenzhou area. The disease was correlated with age and sex, often misdiagnosed and misclassified. Physicians and as well as general publics should be trained to identify the cases. PMID- 15132867 TI - [Development and application of geographic information system of Aedes vector in Chaozhou city, Guangdong province]. AB - OBJECTIVE: To explore the spatial distribution character of dengue fever and the change of Aedes' population, so as to provide macroscopical decision-making evidences of prevention and supervision on dengue fever. METHODS: (1) Collecting data on morbidity of dengue and supervision on vector's population in the corresponding period. (2) Drawing digitized map of Chaozhou in scale of 1:50,000, including elements of boundary, residential areas, road and traffic, altitude, water systems etc. (3) Measuring the latitude and longitude of center position of surveillance safes on the scene. (4) Processing spatial analysis by the ArcGIS 8.5 software. RESULTS: Distribution of Aedes showed spatial cluster in Chaozhou, while its density was related to the distance to the watersides. The closer to the watersides, the higher the density was. Map on spatial distribution showed that although the Aedes epidemic situation changed yearly, but primarily be kept in high, middle, low regions. Cross-validation effects of the distribution maps were satisfactory. CONCLUSION: Geographic information system was promising in analyzing data on dengue fever, and better than other routine research methods. PMID- 15132868 TI - [Impacts of birth defects on perinatal deaths in Chinese population]. AB - OBJECTIVE: To examine the time trends of perinatal mortality and the frequency of birth defects occurring in perinatal deaths, and to provide a national perspective on the impacts of congenital anomalies on perinatal mortality from 1990 through 2001. METHODS: Data were from Chinese Birth Defects Monitoring network-a hospital-based congenital anomalies registry system. During 1990 - 2001, all live or still births with 28 weeks of gestation or more, born in monitoring units, were studied within 7 days after delivery. The proportion of perinatal deaths due to birth defects, which was defined as the number of perinatal deaths associated with congenital anomalies per 100 perinatal deaths, was calculated by birth area (urban versus rural), geographic-economic status (coast areas, inner land areas and remote areas), to evaluate the impacts of birth defects on perinatal mortality. RESULTS: Perinatal mortality declined from 22.85 per 1000 in 1990 to 13.26 per 1000 in 2001, which showed a significant downward trend. Similar trend was also observed in the rate of stillbirth and the ratio of early neonatal death. However, the proportion of perinatal deaths due to birth defects had an increasing trend although the perinatal birth defects specific death rate was declining, especially during 1996 - 2001. This result was also seen in urban and rural area, in coast regions, in inner land regions and in remote regions of China. Higher rate of birth defects occurring in perinatal deaths was observed in urban area than in rural area. Significant difference of this rate was also found among different geographic-economic regions, with the highest one in inland regions. CONCLUSION: Birth defects were accounted for an increasing proportion of perinatal deaths in China, and had become one of the major causes of perinatal deaths. PMID- 15132869 TI - [A baseline survey on the association of smoking onset and life events on adolescents in Wuhan, China]. AB - OBJECTIVE: To understand the association between stressful life event and onset of smoking among adolescents in China. METHODS: Data on baseline survey from a 5 year longitudinal smoking prevention trail was used. Five thousand, five hundred and thirty-six students, from grade 7, 8 and 9 in 22 junior high schools all over Wuhan, China, had completed the survey on the onset of smoking in the past year and on stressful life events. Logistic regression analysis was used. RESULTS: The results showed that 8.2% of the respondents reported the onset of smoking in the past year before the survey. The rates for male and female were 13.6% and 3.4% respectively while 7.8% for 7th graders, 9.1% for 8th graders and 8.3% for 9th graders. When gender, urbanicity and age were controlled with logistic regression model, the onset of smoking in general was significantly positively associated with the negative life events at school and family while negatively associated with positive life events in school and personal domain (P < 0.05). CONCLUSION: Attention should be paid to help adolescents to cope with stressful life events in order to prevent and control the onset of smoking among adolescents. PMID- 15132871 TI - [The anti-respiratory syncytial virus effect of an active compound (AP3) from a Chinese medicinal herb-Herba patriniae in vitro]. AB - OBJECTIVE: To study the effect on anti-respiratory syncytial virus of an active compound (AP3) from a Chinese medicinal herb-Herba patriniae in vitro. METHODS: Active component of herba patriniae (AP3) was extracted and its anti-respiratory syncytial virus (RSV) effect was tested. A water soluble substance (AP3) was isolated from a Chinese herb Herba patriniae, by hot water extraction, ethol precipitation and gel permeation column chromatography. The cytotoxicity of AP3 was tested by adding it to HeLa cells directly. Its effect against RSV was estimated by CPEI assay while ribavirin was used as positive control. RESULTS: Chemical test showed that the nature of substance AP3 was polysaccharide. The median cytotoxic concentration (TC(50)) of AP3 was 11.45 mg/ml by morphological observation and the median effective concentration (50% effective concentration, EC(50)) of it against replication of the long strain of RSV in HeLa cells was 0.0986 mg/ml. The Therapeutic index (TI = TC(50)/EC(50)) of AP3 was 116.12, much higher than the TI of herba patriniae (AP1) (TI = 59.26) and ribavirin (TI = 53.45). Moreover, AP3 gave a dose-dependent response in inhibiting RSV. In the assay, the effect of AP3 against RSV growth was also tested. In addition, the effect of AP3 on virus growth, AP3 inhibited replication of RSV in HeLa cells, when added at 0 h, 2 h, 4 h after virus infection, were also tested. CONCLUSION: This study suggested that the AP3 exerted an obvious inhibitory effect to RSV in HeLa cell culture. This study furnished a reliable evidence for development of a new antiviral drug. PMID- 15132870 TI - [Studies on the infection status of seven species Mycoplasma, three species of Chlamydia, Neisseria gonorrhoeae and Garderella vaginalis in 76 patients with sexual transmitted diseases]. AB - OBJECTIVE: To study the infectious status of seven species of Mycoplasma, three species of Chlamydia, Neisseria gonorrhoeae and Garderella vaginalis in the 76 male sexual transmitted disease (STD) patients in Yangzhou city. METHODS: Twelve species of pathogens including Ureaplasma urealyticum (Uu), Mycoplasma hominis (Mh), Mycoplasma pneumoniae (Mpn), Mycoplasma genitalium (Mg), Mycoplasma fermentans (Mf), Mycoplasma penetrans (Mpe), Mycoplasma prium (Mpi), Chlamydia trachomatis (Ct), Chlamydia pneumoniae (Cpn), Chlamydia psittaci (Cps), Neisseria gonorrhoeae (Ng) and Garderella vaginalis (GV) were detected by nested polymerase chain reaction including PPNG. RESULTS: The positive rates of Uu, Mh, Mpn, Mg, Mf, Mpe, Ct, Ng were 64.5%, 27.6%, 26.3%, 18.4%, 2.6%, 2.6%, 31.6%, 36.8%, in which Penicillinase-producing neisseria gonorrhoeae (PPNG) accounted for 14.3%, GV 15.8%. No Mpi, Cpn or Cps were found. There was more significant therapeutic effects on the detectable rate of Mycoplasma nucleic acid between positive gonococcus and negative gonococcus in male STDs patients (chi(2) = 3.848, P < 0.05). CONCLUSION: The infection rates of Mycoplasma, Chlamydia, Ng and GV were high among male STD patients in Yangzhou city. In clinical practice, more attention should be paid on correct diagnosis and treatment for patients, with Gonococcus, Chlamydia, Mycoplasma and GV. PMID- 15132872 TI - [Detection of Schistosomia japonicum 5D gene by polymerase chain reaction and genechip technique]. AB - OBJECTIVE: In order to develop the diagnostic genechip for specific detection of Schistosoma japonicum (Chinese mainland strain). METHODS: Probe and primers were designed based on the Schistosoma japonicum 5D gene encoding an immunogenic miracidial antigen. The probe for the conservative and specific gene sequence was spotted onto the specially treated glass slides by pin-based spotting robot Pixsys 5500 and was employed to make genechips. A polymerase chain reaction (PCR) protocol was designed to effectively amplify the 5D gene fragment containing the probe sequence from cercaria, egg, adult worm and infected Oncomelania DNA as well as other flukes DNA, respectively. After 35 cycles by PCR, the products were then labeled with fluorescent Cy3-labeled primer, using dissymmetrical PCR. The labeled PCR products of the target genes were hybridized to the diagnostic genechips for detection of Schistosoma japonicum and a fluorescent scanner (ScanArray 3000) was used to observe and record the hybridization signals. RESULTS: The result obtained from the study showed that a 262 bp DNA fragment was amplified from cercaria, egg and adult worm with the designed primers and enable the genechip be applied to detect a single cercaria, egg and adult worm. When the genechip was used to detect Clonorchis sinensis, Fasciolopsis busk, and Paragonimus westermani DNA, the results showed negative, indicating that the genechip had good specificity. CONCLUSION: The genchip technique for detection of Schistosoma japonicum was established successfully and having the characteristics of high sensitivity and specificity. PMID- 15132873 TI - [Study on the relationship between nitric oxide synthase gene G894T polymorphism and hypertension related risk factors in patients with essential hypertension in Chongqing city]. AB - OBJECTIVE: To explore the relationship between genetic anomaly and risk factors in hypertension, a polymorphism at position G894T of the gene encoding the endothelial nitric oxide synthase (eNOS) together with hypertension related risk factors were observed in patients with essential hypertension (EH) in Chongqing city. METHODS: Two hundred and twenty-six patients with EH and matched controls were selected. Genotypes of polymorphisms were determined by polymerase chain reaction (PCR), while PCR products were digested by restriction endonuclease (BanII). Questionnaire referred to life style, dietary, smoking, alcohol consumption, psychological and mental state, waist-to-hip ratio (WHR), etc was administered. RESULTS: There was no significant difference noticed in genotype distribution for the eNOS gene G894T genotype between hypertensive groups and controls, but difference was found among certain related risk factors, such as salt intake, snoring and WHR, etc. Logistic regression analysis showed no association between 894T allele and hypertension. CONCLUSION: Although the polymorphism of eNOS gene G894T did not seem to play an important and direct role in the pathogenesis of EH it might have indirect effects through certain risk factors. PMID- 15132874 TI - [A study on risk factors among 125 type 2 diabetes mellitus with familial tendency]. AB - OBJECTIVE: To study the risk factors regarding heredity and environment in familial incident type 2 diabetes mellitus (DM). METHODS: To compare the difference of environmental risk factors between type 2 DM, impaired glucose tolerance (IGT) and normal persons through study on familial information and environmental risk factors in 125 familial incident type 2 DM in-patients and out patients from 1999 to 2001. Falconer was used to estimate heritability. Penrose was used to study the heredity damagers by polygene analysis. RESULTS: There was a significant constituent ratio diversity (P < 0.01) in triglyceride, body mass index, waist to hip ratio, hypertension history and physical activities history among 3 groups, while no significant diversity in blood lipids and history of coronary heart disease. 83.42% +/- 5.84% heritability of type 2 DM in 125 familial predigree indicated that dominant major gene might exist in these familiar pedigrees. Analysis of polygene in these groups showed type 2 DM might conform to the model of polygene heredity. CONCLUSION: This study suggested that type 2 DM had significant heritability and genetic heterogeneity, which generally appeared to be a disease of multi-factorial inheritance. Environmental risk factors, genetic factors and their interactions were due to type 2 DM. PMID- 15132875 TI - [A randomized controlled Multi-center clinical trial on mosapride in the treatment of functional dyspepsia]. AB - OBJECTIVE: To evaluate the effectiveness and safety of mosapride on treatment of functional dyspepsia. METHODS: Randomized controlled clinical trial was conducted and patients suffered from functional dyspepsia were included. 5 mg mosapride was given three times daily for 4 weeks in the treatment group. 10 mg domperidone was given three times daily for 4 weeks as control. Changes on symptom score, gastric empty or new occurring events were included as outcomes. RESULTS: 231 patients suffered from functional dyspepsia were selected by inclusion and exclusion criteria from August 15 to Oct 22, 1999. Of these, 108 (46.8%) were males, versus 123 (53.2%) females and 118 (51.2%) in the treatment group and 113 (48.9%) as controls. 222 (96.1%) patients were followed up. Results showed that the total efficacy rates in early satiety and abdominal distension were 84.5% and 90.1% in mosapride after the 2 weeks of treatment. Mosapride seemed to be more effective in improving symptoms of belching and heartburn than that in controls (P < 0.05). In 4 weeks, the total efficacy in improving symptoms of abdominal distention and belching showed more effective in mosapride than that in controls (P < 0.05). Decrease of symptoms score was more in mosapride than that in controls (P < 0.05). Mosapride was less effective in controls in improving the gastric empty in terms of proportion (46.2% vs. 25.9%, P = 0.020) and range (46.2% vs. 24.0%, P = 0.003). Side effects would include diarrhea, constipation, headache, dizziness, insomnia, skin scare and the like. There was no significant difference between the two groups (9.6% in mosapride vs. 14.0% in controls). CONCLUSION: Mosapride was safe and effective in improving the symptoms and gastric empty of functional dyspepsia. PMID- 15132877 TI - [Methods for time trend analysis of cancer incidence rates]. AB - OBJECTIVE: To introduce statistical methods of time trend analysis on cancer rates. METHODS: Cancer incidence data collected by the Shanghai Cancer Registry during 1991 to 1999 was used in the analysis to calculate the crude and age adjusted rates, percent changes (PCs) and annual percent changes (APCs). APCs were estimated by a linear regression of the logarithm on the incidence rates during the nine years. It also introduced a method for partitioning a linear trend in age-adjusted rates into site-specific contributions to the overall floating trend. 95% confidence intervals for the APCs and contributions were described in the paper. RESULTS: A decreasing rates were observed for cancers of stomach and esophagus among both men and women in urban Shanghai from 1991 to 1999. The increasing rates among men would include cancers of colon, rectum, gall bladder, pancreas, prostate, urinary bladder, kidney and leukemia. The rates of cancers among women increased for colon, rectum, lung, breast, gall bladder, endometrium, ovary, urinary bladder and kidney. The changes of above cancers over time were statistically significant (P < 0.05 or P < 0.01), but rates for other cancer sites changed little. The APCs (weighted method) and contributions for the cancers of stomach, esophagus, colon, rectum and prostate were -2.99% and 65.72%, -2.90% and -17.07%, 12.30% and 21.46%, 2.94% and 18.62%, and 3.11% and 15.09% among men, and -6.05% and -39.55%, -1.08% and -35.19%, 2.81% and 28.64%, and 3.69% and 15.70% for the cancers of stomach, esophagus, breast and colon in women, respectively. CONCLUSION: APC, and related statistics could be used to describe and analyze the time trend of cancer rates rather than PC or/and graphical method alone. PMID- 15132876 TI - [Alpha-blockers and bioflavonoids in men with chronic nonbacterial prostatitis (NIH-IIIa): a prospective, placebo-controlled trial]. AB - OBJECTIVE: The National Institutes of Health (NIH) category IIIa chronic prostatitis syndromes (non bacterial chronic prostatitis) were common disorders but with few effective therapies. Alpha-blockers and bioflavonoids had recently been reported in randomized controlled trials to improve the symptom of these disorders in a significant proportion of men. The aim of this study was to confirm these findings in a prospective randomized, placebo-controlled trial. METHODS: Forty-five men with category IIIa chronic non bacterial protatitis were randomized into three groups as follows: (1) placebo; (2) phenoxybenzamine hydrochloride:10 mg two times a day for one month; (3) flavoxate HCI-neptumus: 200 mg three times a day for one month. The NIH chronic prostatitis symptom score was used to grade symptoms at the beginning and conclusion of the study. RESULTS: All the patients in three groups completed the study except three dropout patients in placebo group because of sever symptoms. The three groups were similar in age, duration of symptoms and initial symptom score. Patients taking placebo had a mean improvement in NIH-CPSI from 21.85 to 19.55 (not significant), while the phenoxybenzamine-hydrochloride group had a mean improvement from 21.95 to 13.75 (P < 0.01), and those taking flavoxate HCI-neptumus had a mean improvement from 21.75 to 16.95 (P < 0.05). The decrease in NIH-CPSI was associated with significant improvement in patients' clinical manifestations. CONCLUSION: Therapy with alpha-blockers was well tolerated with significant symptomatic improvement in most men having chronic non-bacterial chronic protatitis while the bioflavonoids group had no significant improvement. Mechanism of both medicines needs further study. PMID- 15132878 TI - Prevention of sudden cardiac death and diabetes mellitus. PMID- 15132879 TI - Influence of weight loss and physical activity interventions upon muscle lipid content in relation to insulin resistance. AB - Numerous cross-sectional studies find association between intramyocellular lipid (IMCL) content and insulin resistance (IR). However, less is known concerning the effects of interventions on this association. For instance, the effects on IMCL content by interventions of weight loss and physical activity, classically known to improve IR, have only recently begun to be examined. In a recent study, content of IMCL following bariatric surgery was found to decrease dramatically. The decrease in IMCL content was related to the improvement in IR that occurred following bariatric surgery. Conversely, several studies find that with physical activity, IMCL content typically remains unchanged or increases, even though IR improves. Potential explanations for this difference between weight loss and physical activity are discussed in this article. PMID- 15132880 TI - Beta-cell failure in the pathogenesis of type 2 diabetes mellitus. AB - Type 2 diabetes is the result of a progressive impairment of pancreatic beta-cell function in the setting of worsening insulin resistance. Studies in high-risk populations have demonstrated that during progression to diabetes, beta cells have declining function and lose the first phase of insulin secretion, resulting in less than adequate suppression of hepatic glucose production following meals. In addition, oscillations of insulin secretion become unmatched from their normal coupling with glucose. Several mechanisms are thought to be responsible for impaired beta-cell function, including glucose toxicity and lipotoxicity, and potentially contribute to beta-cell loss. Advances in molecular science have elucidated several cytokines and transcription factors possibly implicated in the loss of beta-cell mass. In the past 15 years, clinical trials have given hope for potential therapies that may either delay or prevent the progression to diabetes. Lifestyle modification and pharmaceutical treatment remain the most promising interventions. PMID- 15132881 TI - Gene expression in the pathophysiology of type 2 diabetes mellitus. AB - Type 2 diabetes mellitus (DM) is characterized by insulin resistance and pancreatic beta-cell dysfunction. In high-risk subjects, the earliest detectable abnormality is insulin resistance in skeletal muscle. Impaired insulin-mediated signaling, gene expression, glycogen synthesis, and accumulation of intramyocellular triglycerides have all been linked with insulin resistance, but no specific defect responsible for insulin resistance and DM has been identified in humans. With recent advances in genomic techniques, it is now possible to assess gene expression patterns in small samples of muscle tissue from metabolically characterized humans. We have applied these techniques to identify genes and pathways potentially important in the pathogenesis of DM. Both DM and the insulin resistance characteristic of "prediabetes" are associated with reduced expression of genes encoding key enzymes in oxidative metabolism and mitochondrial function, and highlight the potential central role for oxidative metabolic pathways in the pathogenesis of type 2 DM. PMID- 15132882 TI - Heart protection study: a focus on diabetic patients. PMID- 15132884 TI - Association of diabetes and hepatitis C infection: epidemiologic evidence and pathophysiologic insights. AB - Up to one third of patients with chronic hepatitis C virus (HCV) develop type 2 diabetes mellitus (DM). This prevalence is much higher than that observed in the general population, and in patients with other chronic liver diseases such as hepatitis B virus, alcoholic liver disease, and primary biliary cirrhosis. Further, HCV seropositivity in patients with DM appears to be higher than in the general population. Post- liver transplantation DM also appears to be higher among patients with HCV. In this article, we review the epidemiologic association between HCV and DM, highlighting the most recent pathophysiologic insights into the mechanisms underlying this association. PMID- 15132883 TI - Sudden cardiac death in patients with diabetes. AB - Sudden cardiac death (SCD) affects over 450,000 people in the United States annually. The mechanisms involved are poorly understood. The predictors currently known include traditional coronary heart disease risk factors, electrocardiographic abnormalities, cardiac autonomic neuropathy, left ventricular hypertrophy, cardiomyopathy, and conduction abnormalities. Diabetes mellitus and impaired glucose tolerance are of special importance due to their increased prevalence reaching epidemic proportions and the elevated risk of SCD in people with these disorders. This article reviews the current predictors of SCD with a focus on people with diabetes, hoping to offer physicians and researchers a better understanding of and a solid ground for further needed research on this important cause of premature death. PMID- 15132886 TI - Improving outcomes in patients with acute coronary syndromes. PMID- 15132885 TI - Management of hypertension in the cardiometabolic syndrome and diabetes. AB - This article reviews the goals of antihypertensive therapy in patients with the cardiometabolic syndrome, as well as diabetes in the context of reducing progression of kidney disease and decreasing cardiovascular (CV) mortality. All published guidelines recommend a blood pressure (BP) goal of less than 130/80 mm Hg in people with diabetes. To achieve this BP, an average of three different antihypertensive agents, appropriately dosed, are needed. Initial therapy includes an inhibitor of the renin-angiotensin-aldosterone system usually coupled with a thiazide diuretic. Beta-Blockers are often employed to both lower BP and reduce overall CV risk; however, nondihydropyridine calcium antagonists are comparable in benefit without the adverse metabolic effects. Changing lifestyle patterns to include exercise and proper diet, achieving target BP and lipid goals, and treating with an aspirin daily reduces the absolute risk of a CV event by 20% over less intensive treatment. Thus, treating the cardiometabolic syndrome requires an aggressive approach with a focus on both lifestyle modification and pharmacologic intervention. PMID- 15132887 TI - Reducing mortality in critically ill patients. PMID- 15132888 TI - Type 2 diabetes in South Asians: a pathophysiologic focus on the Asian-Indian epidemic. AB - There is a high prevalence of type 2 diabetes mellitus and coronary artery disease among urban and migrant Asian Indians, despite the absence of traditional risk factors. Evidence exists that Asian Indians are more insulin resistant than white persons and that insulin resistance may play an important role in the pathogenesis of these diseases. Increased visceral fat in Asian Indians is associated with increased generalized obesity, which is not apparent from their nonobese body mass index. Increased visceral fat is related to dyslipidemia and increased frequency of insulin resistance and may account for the increased prevalence of diabetes mellitus and cardiovascular disease in Asian Indians. In addition, early protein energy deprivation, as indicated by low weight at birth and at 1 year of age, may induce a state of vulnerability to the development of type 2 diabetes in later life, especially if the quantitative and qualitative aspects of nutrition and altered lifestyles during adult years pose an additional challenge. PMID- 15132889 TI - Diabetes in African Americans: unique pathophysiologic features. AB - Type 2 diabetes is an increasing public health problem among African Americans, especially children. Several features make type 2 diabetes among African Americans unique. First, African-American adults with type 2 diabetes, or Flatbush diabetes, present with diabetic ketoacidosis. Patients are insulin resistant with acute, severe defects in insulin secretion and no islet cell autoantibodies. Following treatment, some insulin secretory capacity is recovered and ketoacidosis generally does not recur. The second is remission in African Americans with type 2 diabetes. Recovery of glucose homeostasis, accompanied by recovery of beta-cell function, follows intensive glycemic regulation. Finally, among African Americans with diabetes who are not obese, normal insulin sensitivity is not uncommon. Such individuals do not have the increased cardiovascular risk of insulin-resistant individuals. Differences in visceral, not subcutaneous, adipose tissue volume appear to determine insulin sensitivity. Understanding the unique physiologic and clinical features of African Americans is critical in designing appropriate treatment strategies. PMID- 15132891 TI - STAT1 activation and STAT1-dependent immune-response gene ICAM-1 expression in alveolar macrophages of rats suffered from interstitial pulmonary fibrosis. AB - AIM: To investigate the role of signal transducer and activator of transcription 1(STAT(1)) in alveolar macrophages (AMs) from rats with bleomycin-induced pulmonary fibrosis. METHODS: Fifty adult female Wistar rats were randomly divided into two groups: One group was intratracheally instilled with bleomycin (BLM), while another group with 9 g/L NaCl solution (NS). The kinetic change of STAT(1) activation and intercellular adhesion molecule-1(ICAM-1) expression in AMs was examined by Western blot and immunohistochemical staining, respectively. RESULTS: (1)STAT(1) was rarely activated in AMs of NS group. After bleomycin treatment, the STAT(1) activation of AMs was significantly increased on day 1, climaxed on day 7, and then gradually decreased, but remaining significantly higher than that of NS group on day 28 (P<0.05). (2)There was only a few cells ICAM-1 higher than that positively stained in AMs of NS group. After intratracheal instillation of bleomycin, the number of ICAM-1 positive stained cells was significantly increased on day 1, reached its peak values on day 7, and then gradually decreased, and still higher than that of NS group on day 28 (P<0.05).(3)There was a significant correlationship between STAT(1) activation and ICAM-1 expression in AMs (r =0.913, P<0.01). Further more, the ICAM-1 expression in AMs was significantly correlated with the severity of inflammation in lung tissues (r =0.947, P<0.01). CONCLUSION: STAT(1) was abnormally activated in AMs of rats with bleomycin-induced interstitial pulmonary fibrosis (IPF). The abnormal STAT(1) activation may play a role in the pathogenesis of acute alveolitis and pulmonary fibrosis. PMID- 15132890 TI - Diabetes in Montana's Indians: the epidemiology of diabetes in the Indians of the Northern Plains and Canada. AB - The prevalence of diabetes is two- to threefold higher in American Indians in Montana compared with the non-Indian population. High rates of diabetes have also been described in Canadian aboriginal populations closely related to the tribes in Montana. Diabetes in pregnancy has increased among Indian mothers and high birth-weight babies are increasingly likely to be born to Indian mothers with diabetes in pregnancy. Over 70% of the incident cases of diabetes in youth less than 20 years of age on the reservations have the clinical characteristics of type 2 diabetes. Cardiovascular disease mortality rates are high among Indians in Montana, and the prevalence of smoking in the Indian populations of Montana and the neighboring tribes in Canada is remarkably high. Indians in Montana are more likely than non-Indians of similar age to believe that diabetes is preventable and to recall advice about diabetes risk. PMID- 15132892 TI - [Effect of nitric oxide on NF-kappaB activation induced by cigarette smoke extract in rat alveolar macrophage]. AB - AIM: To explore the effect of nitric oxide on NF-kappaB activation in rat alveolar macrophage(AM) induced by cigarette smoke extract(CSE) and the involved mechanism. METHODS: AMs were co-cultured with NO precursor L-arginine (L-Arg) or inhibitor of inducible nitric oxide synthase L-N(6)(1-iminoethyl)lysine (L-NIL) and CSE. The expression of NF-kappaB was determined by immunocytochemistrical staining, the content of I-kappaB protein was detected by Western blot, the level of NO in supernatant was assayed by Griess reaction. RESULTS: CSE could induce NF kappaB activation and decrease level of I-kappaB. L-Arg augmented CSE-induced NF kappaB activation and decreased I-kappaB content at low concentration while inhibited CSE-induced NF-kappaB activation at high concentration, but there was not any significant difference of the expression of I-kappaB. The percentage of NF-kappaB nucleus dyeing positive cells was lower and the expression of I-kappaB was higher in response to CSE plus various concentrations of L-NIL as compared with that in response to CSE alone (P<0.01). The changes induced by L-NIL were concentration-dependent. CONCLUSION: NO may up-regulate and down-regulate the activation of NF-kappaB induced by CSE according to difference in NO concentration in AMs. PMID- 15132893 TI - [Effect of the truncated human apoptosis-inducing factor expression on apoptosis of HeLa cells]. AB - AIM: To observe the expression of the truncated human apoptosis-inducing factor (AIF) gene and its apoptosis-inducing effect on HeLa cells. METHODS: Full-length human AIF gene was cloned by RT-PCR, then the truncated AIF gene was constructed by deleting the N-terminal mitochondrial location sequence (MLS), and inserted into the EGFP co-expression vector pIRES2-EGFP. After being transfected into HeLa cells with Lipofectamin, the expression of the truncated AIF gene and its effect on HeLa cells morphology and growth condition were detected by fluorescence microscope, immunohistochemical staining, indirect fluoroimmunoassay and electron microscope analysis. RESULTS: The eukaryotic expression vector pIRES2-EGFP containing of truncated human AIF gene was constructed successfully. The AIF protein could be detected in the transfected HeLa cells. After transfection, typical apoptotic feature of the transfected HeLa cells and a mass of cell death were observed under electron microscope. CONCLUSION: The expression of the truncated human AIF gene can induce apoptosis of the transfected human HeLa cells. PMID- 15132895 TI - [Cloning of mouse p27Kip1 and investigation on its role in apoptosis and survival of peripheral blood B lymphocytes]. AB - AIM: To clone the mouse p27(Kip1) gene and analyze its expression and possible function in the signaling through antigen receptor and CD40 in peripheral blood B cells. METHODS: Mouse p27(Kip1)was cloned from cultured B cells by RT-PCR. Apoptosis of the B cells was detected by flow cytometry. Expression of p27(Kip1)gene was detected by Northern blot. RESULTS: Mouse p27(Kip1)gene was successfully cloned. In consistence with B cell apoptosis or survival, cross linking of antigen receptor also induced up-regulation of p27(Kip1)in mouse B cells, which was blocked by CD40 signaling. Antigen receptor-mediated B cell apoptosis was blocked by antisense oligonucleotides directed against p27(Kip1). CONCLUSION: These data suggest that p27(Kip1)may be involved in apoptosis and survival of B cells mediated by antigen receptor and CD40 signals, respectively. PMID- 15132894 TI - [Effect of CD28 costimulator on T cell receptor(TCR)-induced apoptosis of thymocytes]. AB - AIM: To analyze apoptosis and its degree of thymocytes at early and intermediate phases induced by anti-TCRalphabeta mAb or anti-TCRalphabeta mAb+anti-CD28 mAb stimuli, and to analyze the influence of CD28 costimulator on TCR-induced apoptosis of thymocyte subsets. METHODS: Thymocytes were freshly prepared and were cultured for 20 h in the presence of anti-TCRalphabeta mAb+anti-CD28 mAb or anti-TCRalphabeta mAb along, The cultured cells were stained with fluorescein labelled Annexin V, PI, anti-CD4 mAb and anti-CD8 mAb, then color reagents. The apoptotic cells were analyzed by FACS. RESULTS: Compared with the spontaneous apoptosis of thymocytes cultured in medium alone, CD28 costimulator markedly enhanced the number of thymocyte apoptosis at early and intermediate phases; under the action of double signaling stimulators, the apoptosis of CD4(+) CD8(+)(DP) thymocytes were substantially increased, and the expression of CD28 were also upregulated on these apoptotic DP cells. CONCLUSION: Influence of CD28 costimulator on TCR-induced apoptosis of thymocyte subsets might be related to thymocyte's mature degree. Double signaling may induce apoptosis of DP thymocytes. PMID- 15132896 TI - [The expression and application of human Fas ligand in E.coli]. AB - AIM: To express recombinant human FasL molecule in E.coli. METHODS: RT-PCR was applied to amplify FasL cDNA from the total RNA extracted from activated human peripheral blood lymphocytes. The DNA fragment was cloned into PCR2.1 vector. After sequencing, the FasL gene was inserted into pQE-31 vector and expressed in E.coli M15. The FasL protein was purified through Ni-ATA affinity chromatography column and identified by SDS-PAGE and Western blot. The mice were immunized with the FasL protein and the specific anti-serum was harvested 6 weeks after immunization. The serum level of FasL from with different kinds of diseases patients were detected using the anti-FasL antibodies from the immunized mice. RESULTS: The expressed protein could be recognized by anti-human FasL antibody in Western-blot analysis with M(r)40 000. This protein could induce Jurket cells apoptosis. anti-FasL serum prepared from mouse could detect the serum FasL as sensitive as commercial ELISA kits. CONCLUSION: The human FasL protein is obtained. It lays the foundation for the further detecting the concentration of FasL and sFasL of patients. PMID- 15132897 TI - [Anergy, apoptosis and decreased MHC class I molecule expression of C57BL/6J mouse CD4+ T cells induced by SEB]. AB - AIM: To study the mechanism of peripheral T cell tolerance induced by superantigen Staphylococus aureus enterotoxin B (SAgSEB). METHODS: Splenocytes from C57BL/6J (B6) mice were stimulated with SEB in-vitro. The proliferation of lymphocytes was determined by MTT colorimetry. The apoptosis of anergic T cells, the cells being in the S and G(0)-G(1) phase and MHC-I (H-2K(b)) expression of T cells were analyzed by flow cytometry (FCM). Apoptosis of anergic T cells was also demonstrated by DNA electrophoresis ladder. RESULTS: SEB could induce strong proliferation of CD4+ T cells and the maximal percentage of CD4+ T cells in S phase appeared on 3rd day after SEB stimulation and then decreased, but the changes of percentage of CD4+ T cells in G(0)-G(1) phase was opposite to that in S phase. CD4+ T cells became anergy on 3rd day and apoptosis of anergic CD4+ T cells appeared on 5th day. Percentage of apoptosis cells increased gradually and could not be reversed by addition of anti-CD3 Ab and ConA. Expression of MHC-I (H 2K(b)) was marginally declined accompanied by CD4+ T cell anergy after SEB exposure. CONCLUSION: SEB-induced tolerance may have relation to anergy and apoptosis of CD4+ T cells, and down-regulation of MHC-I molecules. PMID- 15132898 TI - [The role of superantigen SEB-induced immunotolerance in the immune privileged site]. AB - AIM: To investigate if the immunological tolerance induced by superantigen staphylococcus enterotoxin B (SEB) takes part in the reaction at the immune privileged site. METHODS: Corneal transplantations were carried out using LEW rats as recipient and F344 rats as the donor. The recipient rats were randomly divided into 5 groups, receiving peribulbar SEB injection of a dosage of 30, 60, 90 or 120 microg/kg 7 days before and after operations. The negative control group received saline injection while the positive control group received glucocorticoid(GC), FK506, CsA or interleukin-1 receptor antagon ist (IL-1ra), respectively. The survival time and rejection index of corneal grafts within 30 days postoperation were examined and recorded using anti-rejection index method. The inflammatory cell infiltration of the recipient cornea and changes of local NK cells were examined by PE-conjugated anti-NK1.1 monoclonal antibody. RESULTS: The grafts in the group receiving injection of 120 microg/kg SEB survived 22 days longer than the negative control group. Their survival time is also significantly longer than the groups treated with FK506, CsA, GC or IL-1ra. The graft rejection index was 3.42+/-2.18 in the rats receiving 120 microg SEB/kg,but 6.58+/-3.15 in the negative control group (P<0.01). The edema and new blood vessel indexes were also significantly reduced. The immunostaining showed that the number of NK cells increased following injection of SEB. CONCLUSION: Injection of SEB reduced the rejection to corneal transplants in rats, showing that the immune tolerance induced by superantigen SEB plays a role in the reaction at the immune privileged site. PMID- 15132899 TI - [The effect of newcastle disease virus on the biological behavior of tumor cells]. AB - AIM: To investigate the anti-tumor effect of newcastle disease virus (NDV). METHODS: Plaque formation test was used to investigate the effect of NDV on chicken embryofibroblasts(CEF). Cell suppression test, agarose gel electrophoresis, cytoskeleton staining, fluorescence staining, TUNEL staining, and sialic acid content determination were used to observe the influence of NDV on several human tumor cells. RESULTS: The Plaque formation was observed in chicken embryo fibroblasts.NDV could lead to the apparent cytopathy of BHK, Hela and Hep-2 tumor cells, but has no apparent effect on Wish cells. The strong suppressive effect of NDV on the growth of these tumor cells was found without dose dependence. The optimal dose of NDV could induce the death of tumor cells which was mainly apoptosis, as showed by the classical DNA ladder in DNA gel electrophoresis. NDV also resulted in the changes of cytoskeleton and decreased the level of sialic acid contents on tumor cells. CONCLUSION: NDV may be a potential anti-tumor agent. PMID- 15132900 TI - [Construction and expression of eukaryotic expression vector containing hVEGF165 gene in rat bone marrow stroma cell]. AB - AIM: To construct the eukaryotic expression vector containing human vascular endothelial growth factor 165(VEGF165) gene and express it in rat bone marrow stroma cells(rMSCs). METHODS: The recombinant plasmid pSP73-VEGF165 was digested with BamH I and Xho I. Then the hVEGF165 gene segment obtained was again cloned into pcDNA3.1 to construct recombinant eukaryotic expression vector pcDNA3.1 VEGF165. Then the recombinant vector was identified by enzyme digestion analysis and sequencing. The rMSCs were transformed by recombinant vector and positive clones were screened with G418. The expression of hVEGF165 gene in the transformed cells was detected by immunocytochemical staining. RESULTS: Enzyme digestion analysis and sequencing showed that target gene had been cloned into recombinant vector. The expression of hVEGF165 gene in the transformed cells had been demonstrated by immunocytochemical staining. CONCLUSION: The recombinant eukaryotic expression vector has been constructed and expressed successfully in the transformed cells. Therefore, it is possible to use the rMSCs expressing hVEGF165 gene as seed cells in the bone tissue-engineering. PMID- 15132901 TI - [Expression of PTEN protein in oromaxillofacial tumor cell lines]. AB - AIM: To investigate expression of PTEN protein in oromaxillofacial tumor cell lines and to compare the difference of PTEN protein expression between highly metastatic cancer cell lines and their parent cell lines. METHODS: ABC immunohistochemical staining was applied for detecting expression of PTEN protein in 8 kinds of cancer cell lines, including tongue cancer cell lines Tca8113 and HSC-3,oral bottom cancer cell line HSC-2, buccal cancer cell line BcaCD885, mucoepidermoid carcinoma cell line MEC-1, adenoid cystic carcinoma cell line SACC83, highly metastatic tongue cancer cell line Tb-TLP and highly metastatic mucoepidermoid carcinoma cell line M3SP2. RESULTS: Of the 8 kinds of cancer cell lines, 5 kinds of cancer cell lines were positive for PTEN protein expression. PTEN protein was located in cytoplasm around nucleus. 3 kinds of cancer cell lines, BcaCD885, Tb-TLP and M3SP2, lacked expression of PTEN protein. CONCLUSION: Deficiency of PTEN protein expression may play a certain role in cancer cell metastasis. PMID- 15132902 TI - [Expression and significance of heat shock protein 70, 90 in endometrial carcinomas]. AB - AIM: To explore the expression and significance of heat shock protein (HSP)70, 90 in endometrial carcinomas. METHODS: Expresssions of HSP70 and HSP90 in 30 normal endometrium, 30 endometrial hyperplasia and 53 endometrial carcinomas were detected by Envision immunohistochemical staining and imaging pattern analyser. RESULTS: Gray Scale value(GS) of HSP70 in endometrial carcinoma being (209.06+/ 5.36) GS was obviously higher than that in normal endometrium (145.21+/-4.09)GS, P<0.01 and endometrial hyperplasia(148.59+/-4.23)GS, P<0.01. GS of HSP90 in endometrial carcinoma being (166.98+/-5.71)GS was markedly lower than that in normal endometrium (208.57+/-3.14)GS,P<0.05 and endometrial hyperplasia (249.73+/ 4.94)GS, P<0.01. Expression of HSP70 increased with increased pathological grading of tumor P<0.01, whereas HSP90 expression decreased with increased pathological grading P<0.01. Expression of HSP70 in nonendometrioid carcinoma (229.90+/-3.77)GS was higher than in endometrioid carcinoma (198.37+/ 3.15)GS,P<0.01 and the expression pattern of HSP90(140.21+/-3.22)was contrary to that of HSP70 (176.59+/-2.79)GS,P<0.01. There was no correlations among expression of HSP70, 90 in endometrial carcinoma and depth invaded into muscular layer, clinical stage and lymph node metastasis. CONCLUSION: Both HSP70 and HSP90 may have relation to the genesis and prognosis of endometrial carcinoma. PMID- 15132903 TI - [Study of HLA molecules and its associated genes expression in human ovarian cancer cells]. AB - AIM: To explore the correlation between the expressions of HLA gene and its related gene, and expression of class II transactivator (CIITA) gene induced by IFN-gamma in human ovarian cells. METHODS: The expression of HLA molecule in the tumor cells was detected by Western blot, immunohistochemical staining and flow cytometry. The expressions of transporters associated with antigen processing (TAP), low molecular weight peptides (LMP), and CIITA genes were analyzed by RT PCR. RESULTS: Abnormal expression rate of HLA class I molecule reached 45% in the 11 ovarian cancer cells. The expressed abnormalities of HLA class I molecule had relation to those of 4 genes (TAP1, TAP2, LMP2 and LMP7). Expression of HLA class II molecule was identical with that of CIITA gene in the ovarian cancer cells or other tumor cells. After IFN-gamma treatment, expressions of HLA class I and II molecule were increased in the cells of CIITA-expressing constitutively or inducibly, while no enhancement of HLA molecule expression manifested itself in the tumor cells of that CIITA was not yet expressed after IFN-gamma induction. CONCLUSION: Deletion of TAP and LMP gene expression in the ovarian cancer cells is an important factor causing abnormal expression of HLA class I molecule, suggesting that the CIITA gene has involved in the modulation of HLA classes I and II molecule expressions in the tumor cells. PMID- 15132904 TI - [Construction and biological activities of CTLA4Ig adenovirus vectors]. AB - AIM: To construct CTLA4Ig adenovirus vectors (AdCTLA4Ig) by homologous recombination and study their activity, and to employ the vectors to induce cardiac transplantation tolerance by gene therapy. METHODS: CTLA4Ig gene was cloned to pCA13 adenovirus shuttle plasmid by recombination strategy. Construction of CTLA4Ig adenovirus vectors was performed by homologous recombination of pCA13 plasmid containing CTLA4Ig gene with adenovirus helper plasmid, followed by packaged with 293 cells. Expression and secretion of CTLA4Ig was confirmed by RT-PCR, SDS-PAGE and Western blot. The inhibitory effect of supernate of 293 cells infected with AdCTLA4Ig on MLR in-vitro was observed. A biological activity of CTLA4Ig adenovirus vectors was determined by AdCTLA4Ig gene therapy in rats in-vivo. RESULTS: The Construction of CTLA4Ig adenovirus vector was successful. It was confirmed that the supernatant of 293 cells infected with AdCTLA4Ig could inhibit MLR in-vitro. It was also showed that CTLA4Ig adenovirus vectors could induce transplantation tolerance and prolong allograft survival when they were administrated in rats in-vivo. CONCLUSION: CTLA4Ig adenovirus vectors successfully constructed can infect 293 package cells and secrete CTLA4Ig. The CTLA4Ig protein can inhibit T cell activation. The CTLA4Ig adenovirus vectors can be employed to gene therapy in-vivo, and induce transplantation tolerance. PMID- 15132905 TI - [Construction and expression of prokaryotic expression vector for pTAT-HBV targeted ribonuclease fusion protein]. AB - AIM: To construct Tat-HBV targeted ribonuclease fusion protein prokaryotic expression vector and express it in E.coli. METHODS: The cDNAs encoding HBV targeted ribonuclease, human eosinophil-derived neurotoxin and HBV core protein were respectively cloned into prokaryotic expression vector pTAT-HA. Recombinant plasmids were transformed into E.coli BL21(DE3) LysS, then the transformed cells were induced with IPTG. The expression of the fusion proteins were analyzed by SDS-PAGE and Western blot. RESULTS: The three recombinant plasmids were constructed and expressed after IPTG induction successfully. CONCLUSION: The obtained Tat-HBV targeted ribonuclease fusion protein has laid the foundation for using TR in therapy of HBV infection. PMID- 15132906 TI - [The effect of calcium ionophore on dendritic cells derived from peripheral blood mononuclear cells]. AB - AIM: To explore the effect of calcium ionophore (CI) on dendritic cells (DC) derived from peripheral blood monocytes. METHODS: Peripheral blood monocytes from healthy donors were treated with 100 microg/L rhGM-CSF, 100 microg/L rhGM-CSF plus (10 microg/L) CI, and 100 microg/L rhGM-CSF plus (100 microg/L) respectively. After cultivation of 40 hours, cellular morphology was observed under light microscope and electron microscope. Surface markers on treated PBMCs were analyzed by flow cytometry. MTT colorimetry was used to detect proliferation of autologous T cells. RESULTS: Peripheral blood monocytes treated with 100 microg/L rhGM-CSF plus 100 microg/L CI for 40 hours showed typical morphology of DCs. Simultaneously there was a decrease in CD14 expression and increase in HLA DR, CD40, CD83 and CD86 expressions on these cells. In addition, PBMCs treated with 100 microg/L rhGM-CSF pass 100 microg/L CI for 40 hrs. Could evidently stimulate proliferation of autologous T cells. CONCLUSION: CI can markedly enhance transformation of peripheral blood monocytes induced by GM-CSF into DCs. PMID- 15132907 TI - [Effect of PD98059 on Ras-MAPK signal transduction pathway of chronic myelogenous leukemia]. AB - AIM: To study the effect and mechanism of PD98059 on Ras-MAPK signal transduction pathway of chronic myelogenous leukemia. METHODS: K562 cell line was treated by PD98059, cell viability, DNA synthesis, colony formation and MAPK activity of the treated cells were analyzed. RESULTS: The cell viability, DNA synthesis, colony formation and MAPK activity were significantly inhibited by PD98059 (P<0.05), and the inhibitory effect was dose dependent. CONCLUSION: The inhibitory effect of PD98059 was achieved by blocking Ras-MAPK signal transduction pathway which can become a new target in the treatment of CML. PMID- 15132908 TI - [Modulation of the rate of CD158a+/b+ cells by Th1-and Th2-like cytokines]. AB - AIM: To explore modulation of CD158+ cells in human peripheral blood by Th1-and Th2-like cytokines and provide basic data for inducing immune tolerance and preventing graft-versus-host disease (GVHD) in stem cells transplantation. METHODS: Peripheral blood mononuclear from healthy adults were cultured with Th1 like cytokines IL-2, IFN-gamma and Th2-like cytokines IL-4, IL-6 for 72 hours, rates of CD3+, CD4+, CD8+ cells, CD16+ CD56+ cells and CD158a+/b+ cells were analyzed by FACS. RESULTS: (1) The effects of cytokines on CD3+, CD4+, CD8+ and CD16+ CD56+ cells: the rates of above cells were greatly increased after being treated with IL-2 or IFN-gamma(P< 0.05), but efficacy of IL-2 was higher than that of IFN-gamma(P< 0.05). The rates of above cells in IL-2+IFN-gamma treated cells were higher than that in IL-2 or IFN-gamma treated alone. The rates of above cells were greatly decreased after being treated with IL-4+IL-6(P< 0.05), but efficacy of combination of IL-2+IL-4 was higher than that of IL-4 alone, lower than that of IL-2 alone (P< 0.05). (2) The effects of cytokines on CD158a+/b+ cells: the rates of CD158a+/b+ cells in total mononuclear and in CD3+, CD4+, CD8+ and CD16+ CD56+ cells were significantly raised after being treated with IL-2 (P< 0.01), but had no significance changes after being treated with IFN gamma. The rates of CD158a+/b+ cells were decreased after being treated with IL 4+IL-6, whereas increased after being treated with IL-2+IFN-gamma(P< 0.05), but efficacy of being treated with IL-2+IL-4 was lower than that with IL-2(P< 0.05). CONCLUSION: IL-2 plays an important role in the regulation of CD158a/b expression or proliferation of CD158a+/b+ cells. It may involve in controlling NK cells and T cells activity via expression of regulating these molecules or stimulating proliferating of CD158a+/b+ cells. IL-4 and IL-6 have a slight ability to decrease the rates of CD158a+/b+ cells and IL-4 can partially reverse the effect of IL-2 on CD158a+/b+ cells. PMID- 15132909 TI - [The therapeutic efficacy of tumor-derived heat shock protein70 combining interleukin-2 on tumor-bearing mice]. AB - AIM: To investigate the therapeutic efficacy of the combination of tumor-derived HSP70 and IL-2 on tumor-bearing mice. METHODS: HSP70 was purified from the murine tumor cell line by liquid chromatography. And then identified by SDS-PAGE and Western blot,the purity of purified product was analyzed by capillary electrophoresis. The anti-tumor efficacies of mono-administration of HSP70 or IL 2 and their combination were observed by zoopery. RESULTS: Combination of HSP70 and IL-2 was more effective than either of the two therapeutic agents alone in treating tumor-bearing mice. 10 microg HSP70 displayed obviously therapeutic effect no matter applied alone or combined with IL-2. The averaged life span of the mice treated with IL-2 was only 36.6+/-13.0 days, while 40% of the mice treated with 10 microg HSP70, tumors disappeared eventually and their survival time was more than 90 days, and the averaged life span of this group was over 59.2+/-29.6 days. HSP70 plus IL-2 could get tumors disappeared completely in 60% of mice, averaged life time of this group was over 70.8+/-26.5 days. CONCLUSION: The Combination of HSP70 and IL-2 demonstrated apparently therapeutic effect on tumor-bearing mice. The results will be of greater value to the study on the immunotherapy of human malignant tumors. PMID- 15132910 TI - [Significantly improvement of antibody expression level in CHO cells through downregulation of the DHFR gene in expression vector]. AB - AIM: To increase the expressed level of a human-mouse chimeric antibody against human bladder tumor in dihydrofolate reductase (DHFR) defective CHO cells(CHO/DHFR) via weakening the transcription of DHFR gene in the vector. METHODS: A series of chimeric antibody expression vectors with different deletions and mutations in the modulator sequence of DHFR gene were constructed to downregulate the DHFR gene expression. The vectors were used to transfect CHO/DHFR cells and the transfected cells were subjected to gene amplification in medium containing gradually increasing methotrexate (MTX). The expressed chimeric antibody was quantitated by ELISA. RESULTS: The downregulation of vector-produced DHFR gene introduced by mutation of the modulator sequence could significantly improve the gene amplification effect and the increased antibody production correlated to the reduction of DHFR gene expression. From the best group, a clone with antibody production of 55 microg/(10(6) cells.24 h) was obtained by subcloning. More than 100 microg/(10(6) cells.24 h) was achieved by zinic ion induction. CONCLUSION: MTX induced-increase of recombinant antibody production in CHO cells can be increased by weakening the expression of DHFR gene. PMID- 15132911 TI - [Identification of HCV antigen epitopes from a random 15-peptide library by anti HCV polycloral antibody]. AB - AIM: To analyze epitope recognized by anti-HCV antibodies; patients suffered from hepatitis C. METHODS: Anti-HCV Abs were purified from the patients serum through an affinity chromatography column which was prepared with sepharose 4B coupled with protein A. These Abs were used for biopanning of a phage-displayed random 15 peptide library. RESULTS: After 3 rounds of biopanning, the ratio of output to input increased to 3.3 x 10(3) and the false positive rate reduced to 0.2%, suggesting that the enrichment was effective. After the third round of biopanning, sixteen clones were selected to conduct binding test to Abs from the patients and normal person's sera. Nine of them were proved to react specifically to the sera from the patients. From the deduced insert sequence in the coat protein VIII, the core sequence of WPWS was found in 8 clones. The positive phage clones could react to different patients' and not react to normal person's sera. CONCLUSION: These findings indicate that WPWS motif in the short peptide may mimic the HCV epitope recognized by anti-HCV Abs. PMID- 15132912 TI - [Immunopotency of the recombinant urease B subunit vaccine of Helicobacter pylori after intranasal administration to mice]. AB - AIM: To observe the immunopotency of the recombinant urease B subunit (rUreB) of Helicobacter pylori after intranasal administration to mice. MDTHODS: BALB/c mice were immunized intranasally with rUreB of 20 microg,10 microg and rUreB plus different adjuvants, such as cholera toxin B subunit (CTB), Escherichia coli heat labile enterotoxin B subunit (LTB) and carbopol respectively, four times at an intervals of 7 days. The serum and washing solution from gastric, intestinal, nasal and tracheal mucosas were collected in 7 days after final immunization. IgG and IgA antibodies specific for rUreB were detected by ELISA. RESULTS: The levels of IgA and IgG antibodies in sera every groups of mice immunized intranasally were significantly increased compared with control group (P<0.01). Only the levels of serum IgG of mice immunized with 20 microg dose were higher than those of mice immunized with 10 microg dose. Carbopol could enhance the level of IgA antibodies in washing solution from mouse gastric mucosa after intranasal immunization. The efficacy of LTB as a nasal mucosal immuno-adjuvant was stronger than that of CTB. CONCLUSION: CTB, LTB and carbopol can play the role of adjuvant in nasal mucosal immunization. Intranasal immunization with rUreB can induce not only serum IgG antibody production but also antibody responses of different mucosa. Thus intranasal inoculation is a convenient, effective and cheap immunization way. PMID- 15132913 TI - [Construction and expression of mouse-human chimeric Fab fragment gene of monoclonal antibody SZ-2 against platelet]. AB - AIM: To reduce immunogenicity of a monoclonal antibody SZ-2 specific for human platelet. METHODS: Reverse transcription and polymerize chain reaction were used to amplify the variable region genes of monoclonal antibody SZ-2. The cloned V(H) and V(L) genes were sequenced and fused to human IgG1 constant region gene CH1 and Ckappa in plasmid pSW1. The recombinant plasmid were transformed into E. coli. The expressed recombinant proteins were analysed. RESULTS: The V(H) and V(L) genes were homologous with the published gene sequences of mouse antibody variable region. The concentration of chimeric Fab fragment in expression supernatant was about 180 microg/L detected by ELISA. Western blot analysis showed that SZ-2 Fab/Hu maintained the binding activity to human platelet GPIb. The recombinant proteins could suppress platelet aggregation induced by Ristocatin. CONCLUSION: The variable region genes of SZ-2 are cloned and the mouse-human chimeric Fab fragment is expressed successfully in E. coli. PMID- 15132914 TI - [The preparation of human anti-HBs Fab fragment by bioengineering technique]. AB - AIM: To prepare human anti-HBs Fab by bioengineering technique. METHODS: The specific human anti-HBs Fab gene screened from combinatorial library was cloned into plasmid pBAD/g IIIA, then positive clone was transformed into E.coli Top10. After the fermentation and expression processes, the soluble Fab fragment in the periplasm were purified by Ni-NTA-agarose affinity chromatography,and the inclusion bodies were in turn denatured, solubilized, purified and renatured. The specificity of Fab protein was confirmed by Western blot, and binding activity to HBsAg was verified by Dot blot. RESULTS: The quantity of soluble Fab protein purified from periplasm with Ni-NTA-Agarose which possessed good specificity as well as excellent binding activity to antigens was up to 80 mg per liter, but the biologically active protein acquired after renaturation of the inclusion bodies was quite small. CONCLUSION: Using pBAD/g IIIA-Top10 expression system, the soluble Fab protein with biological activity could be produced from periplasm of the E.coli Top10, and the strategy is available to prepare human anti-HBsAg Fab fragments in large quantity by gene engineering technique. PMID- 15132915 TI - [Preparation and identification of phage-displayed ICAM-1-mimic peptide]. AB - AIM: To obtain bioactive ICAM-1 mimetic peptide. METHODS: Phages displaying P1 and P2 were prepared by phage amplication and PEG precipitation. The binding between phage-displayed peptides and anti-ICAM-1 mAb 15.2 was evaluated by sandwich ELISA and competitive ELISA. Bioactivities of P1 and P2 was detected by immunohistochemical staining. RESULTS: Phage-displayed peptides P1 and P2 could specifically bind to mAb 15.2, and the binding could be competitively inhibited by ICAM-1. Immunohistochemical staining showed that P1 and P2 could mimic the binding of ICAM-1 to its receptor LFA-1. CONCLUSION: Phage-displayed peptides P1 and P2 are bioactive just as native ICAM-1. PMID- 15132916 TI - [Preparation and identification of monoclonal antibodies against CD100 molecule]. AB - AIM: To prepare and identify the monoclonal antibodies (mAbs) against CD100. METHODS: BALB/c mice were immunized with eukaryotic expressed CD100 protein. The splenocytes from immunized mice were fused with Sp2/0 cells and the positive hybridoma clones were screened by indirect ELISA and mAbs reacting to CD100 antigen were identified by indirect immunofluorescent staining and flow cytometry with human acute T lymphoblastic leukemia cell line MOLT-4, activated human PBMC, gibbon T lymphocyte cell line MLA-144 and monkey EBV-transformed B lymphoblast cell line(BLCL). RESULTS: Seven hybridomas secreting mAbs against CD100 molecule were obtained. All the mAbs could react to natural CD100 expressed on human MOLT 4 and activated PBMCs as well as monkey BLCL cells, and six out of them could react to gibbon MLA-144 cells. CONCLUSION: A set of mAbs recognizing CD100 molecule expressed on human, monkey and gibbon cells are obtained, which may provide an useful tool for studying the structure-function relationship of CD100 molecule of human, as well as gibbon and monkey. PMID- 15132917 TI - [Preparation and characterization of monoclonal antibodies against human laminin]. AB - AIM: To prepare monoclonal antibody(mAb) against humen laminin(LN) and identify their properties. METHODS: BALB/c mice were immunized with humen LN, and then mAb was prepared by hybridoma technique. The titres of mAbs in ascetic fluid and relative affinity of mAb were determined by indirect ELISA test. Ig subgroup, specificity and combine dot of mAb were analysed by ELISA. RESULTS: 4 hybridoma cell lines 2A3, 2C6, 3G7 and 4H2 secreting anti-humen LN mAb were obtained. Their Ig subgroup belonged to IgG1, light chains were type kappa, The titres of 4 mAbs in ascitic fluid were 3.6 x 10(4)-2.1 x 10(6). Relative affinity of mAb 2C6 was above 10(12) and relative affinity of other 3 mAbs 2A3, 3G7 and 4H2 was 10(6), 10(8) and 10(8). Epitope recognized by 2 mAb(2A3 and 4H2) differed from the that by 2C6 and 3G7 another. CONCLUSION: 4 mAbs against humen LN has been prepared successfully and possess higher titer, specificity and relative affinity. Preparation of anti-human LN mAbs provide a useful reagent for further studying the role of LN in some diseases. PMID- 15132918 TI - [Purification by FPLC and characterization of Mycobacterium tuberculosis heat resistant peptide antigen stimulating proliferation of human gammadelta+ T cells]. AB - AIM: To purify Mycobacterium tuberculosis heat-resistant peptide antigen(Mtb-Ag) that can stimulate gammadelta+ T cells. METHODS: Mtb-Ag was first separated by fast performance liquid chromatography (FPLC) with S-100 column, and then the components including low molecular weight peptide antigens (Mtb-LW-Ag) peaks were purified by FPLC with Mono Q column. Activity of the purified Mtb-LW-Ag that stimulates human gammadelta+ T cell proliferation was examined by flow cytometry. RESULTS: Mtb-Ag was separated into four peaks(A, B, C and D) by FPLC S-100 column and the peak B, C and D contained Mtb-LW-Ag. The peak B was further separated into six main peaks(B-I-VI), peak C only one main peak(C-main), and peak D eight main peaks(D-I-VIII) by FPLC Mono Q column. Furthermore, peak B, C, B-III and C main peptide could significantly stimulate human gammadelta+ T cell proliferation in a dose of 0.1 mg/L. CONCLUSION: Varied Mtb-LW-Ag peptides are purified from Mtb-Ag by FPLC, and the B-III and C-main peptides may be the main components of stimulating human gammadelta+ T cell proliferation. PMID- 15132919 TI - [Protective efficacy of DNA vaccines encoding mycobacterium tuberculosis Ag85B protein]. AB - AIM: To explore protective efficacy of pTB30m and pTB30s encoding Ag85B protein against infection with M. tuberculosis H (37)R (v). METHODS: BALB/c mice were infected intravenously with 5x10(5) CFU of M. tuberculosisH (37)R (v) six weeks after the last vaccination of pTB30m and pTB30s. At the same time, normal BALB/c mice were injected intravenously with 5x10(6) T cells separated from immunized BALB/c mouse spleen through nylon wool column, and challenges were injected with 10(5) CFU of M. tuberculosis immediately intravenously. After 4 weeks, the CFU in spleens of infected mice were counted respectively. RESULTS: Vaccination with pTB30m and pTB30s produced significant protection against M. tuberculosis proliferation in the mouse spleens following challenge. As compared with the saline-injected mice, CFU in spleens of the mice vaccinated with pTB30m or pTB30s were reduced significantly, being 0.645(log(10) CFU, P<0.01) and 0.839(log(10) CFU, P<0.001), respectively. In contrast, bacterial load in the mice spleens vaccinated with empty plasmid only had little reduction. After adoptive immunization by T cells from the mice vaccinated with pTB30m and pTB30s, the mice might induce partial protection against M. tuberculosis proliferation in the mouse spleens following challenge. CONCLUSION: Protective efficacy of the mice immunized with pTB30s against challenge with M. tuberculosis virulent strain was better than that immunized with pTB30m.Thus, pTB30s is a promising DNA vaccine with respect to the prevention and treatment of tuberculosis. PMID- 15132920 TI - [Effect of overall alkali of Tongbiling on CD69 expression activated mouse T lymphocytes]. AB - AIM: To investigate the effect of overall alkali of Tongbiling(TBL) on CD69 expression on activated mouse T lymphocytes and its possible mechanism. METHODS: Phorbol 12,13-dibutyrate(PDB) or concanavalin (ConA) were added successively into mouse lymphocytic culture with various concentration of overall alkali TBL. After 24 hours, CD69 expression rate on mouse T lymphocytes activated with PDB or ConA was analyzed by flow cytometry. RESULTS: Overall alkali TBL could significantly down-regulate CD69 expression in a dose-dependent manner. CONCLUSION: Overall alkali TBL can significantly inhibit CD69 expression on activated mouse T lymphocytes. This study provided an experimental basis for application of overall alkali TBL to treatment of rheumatoid arthritis. PMID- 15132921 TI - [Expression of CGRP, ACTH and 5-HT in normal human lymph nodes]. AB - AIM: To detect the expression of calcitonin-gene-related peptide (CGRP), adrenocorticotropin (ACTH), and serotonin (5-HT) in normal human lymph nodes and explore their influences on immune system. METHODS: The expression of CGRP, ACTH and 5-HT in the lymph nodes, including proliferative lymph nodes, were detected by SABC Immunohistochemical staining. RESULTS: In 35 cases of lymph node specimens, the positive rates of CGRP, ACTH and 5-HT expression were 88.5%(31/35), 85.4%(30/35) and 94.2%(33/35) respectively. These 3 molecules had a similar distribution in germinal center of lymph nodes, mainly in the cells of paracortical region, the cells in germinal center of lymph follicles and macrophages, etc. CONCLUSION: There are considerably high positive rates of CGRP, ACTH and 5-HT in human peripheral lymph nodes, and their distribution area are basically coincident with that of immunocytes in the lymph nodes. PMID- 15132922 TI - [DNA family shuffling of tissue-type plasminogen activator genes]. AB - AIM: To probe the possibility of screening tPA with higher activity through DNA shuffling. METHODS: Human, rat and rhesus monkey tPA cDNAs were used as the templates of "DNA family shuffling". The diverse tPA library was expressed in CHO cells, then the screening was carried out. RESULTS: Two shuffled clones, t9 and t17 were selected. The activity of tPA expressed by t9 clone was 4-fold higher than the activity of human tPA. tPA expressed by t17 clone was found having a 88 amino-acid deletion, but still showed the same activity as human tPA. DNA sequencing demonstrated that the sequences of t9 and t17 were mainly derived from human and monkey tPA cDNAs. CONCLUSION: Above work has laid the foundation for further experiments in the screening for higher activity of tPA. PMID- 15132923 TI - [Modulatory effect of bisbenzamidine, a specific inhibitor of tryptase on mast cell secretion]. AB - AIM: To study the ability of bisbenzamidine, a specific inhibitor of tryptase, to stabilize mast cells. METHODS: Human tonsil tissues were chopped up finely and digested with collagenase and hyaluronidase. Dispersed cells were separated from undigested tissue by filtration through a nylon gauze. Prior to challenge, the cells were resuspended in complete HBSS and aliquots of 100 mL containing 4 x 10(3)-6 x 10(3) mast cells were added to a 100 microL aliquot of anti-IgE, calcium ionophore, bisbenzamidine or complete HBSS and incubated for 15 min at 37 degrees Celsius. The reaction was terminated by addition of 150 microL ice cold HBSS and the tubes centrifuged immediately. Tryptase was quantified by ELISA. RESULTS: At concentrations of 1 mg/L and 10 mg/L, bisbenzamidine could inhibit spontaneous tryptase release by 47% after 45 min. A 3.2 or 2.6 fold increase in tryptase release was achieved when cells were incubated with 1 g/L anti-IgE antibody or 1 micromol/L calcium ionophore for 15 min at 37 degrees Celsius. A dose dependent inhibition of tryptase release induced by anti-IgE antibody was observed when dispersed tonsil cells were treated with bisbenzamidine. CONCLUSION: Bisbenzamidine can inhibit IgE-dependent tryptase release from human tonsil mast cells. Therefore, it seems likely that this inhibitor of tryptase is a novel promising mast cell stabilizing drug for treating and preventing allergic inflammation or other mast cell associated diseases. PMID- 15132924 TI - [Induction of IgG and IgM production by Epstein-Barr virus protein]. AB - AIM: To detect the production of IgG and IgM by cultured umbilical blood B cell stimulated by UV and heat-inactivated Epstein-Barr virus (EBV). METHODS: Mononuclear cells were isolated routinely from umbilical blood, then monocytes, NK cells and cytotoxic T cells were eliminated by L-leucine methy ester method; remained T cells were eliminated by AET-SRBC rosette method. The purified B cells were treated with UV or heat-inactivated EBV respectively and cultured in complete IMDM. The IgG and IgM in the supernatant were detected by ELISA. RESULTS: The IgG and IgM production(A value) in UV-inactivated EBV group increased from 18th day to 30 th day after stimulation compared with the baseline. The A value of all time points in heat-inactivated EBV group had no significant difference. CONCLUSION: UV-inactivated EBV may induce IgG and IgM production with a time-effect relationship, suggesting that EBV protein is a key component of stimulating Ig production. This result provide the basis for the further researching the effects of functional EBV protein on the production of natural autoantibodies (NAA). PMID- 15132926 TI - Challenges in total ankle arthroplasty. AB - In the past, total ankle arthroplasty was largely abandoned due to poor survivorship most often caused by loss of bone support. High complication rates were also reported. Despite this, there is renewed interest in ankle arthroplasty and encouraging results are seen in survivorship with midterm follow-up. The procedure, however, remains more challenging than total hip or total knee arthroplasty. With the limited soft tissue envelope, wound problems are not uncommon. Forces at the ankle are very large and yet the surface area for prosthetic support is small. Therefore, fixation can be more difficult. The strongest bone can be eccentric at the distal tibia. The tibial prosthesis can, therefore, tend to settle into the softer bone often laterally. Polyethylene needs to be sufficiently thick to maintain its integrity but that requires a larger bone resection, which weakens bone support. Polyethylene failure or wear leads to the majority of failures in hip and knee arthroplasty. There is a need for further basic science research in total ankle arthroplasty. The lessons learned from other arthroplasty should be considered in ankle arthroplasty design. PMID- 15132927 TI - Bone mineral density, gait analysis, and patient satisfaction, before and after ankle arthroplasty. AB - Bone mineral density (BMD) and patient satisfaction were measured and gait analysis was performed in patients treated with unilateral dual-coated ankle arthroplasty. The study comprised 14 patients (eight women and six men) measured preoperatively and at a median of 15 (range, 12-26) months after surgery. BMD was measured bilaterally in the distal tibia and in the calcaneus. A plantar pressure analysis was performed, including foot contact duration and a new index for describing the ground reaction force curve called the Valley Index (VI). The patients also evaluated their level of pain, ability to cope with daily activities, adaptation of shoes, and walking ability before surgery and at follow up on a visual analogue scale (VAS). Bone mass significantly increased adjacent to the tibial part of the prosthesis. The patients experienced less pain and improved walking and performance in daily activities. There was also a decrease in foot contact duration as well as an increase in VI and ankle range of motion. Change in bone mass was the variable that correlated best with the patients' evaluation of the general outcome of ankle arthroplasty. Correlations among other variables are described. The study concluded that a fairly simple analysis of the ground reaction force curve, bone mass measurements, and VAS can demonstrate that ankle arthroplasty is able to normalize gait, decrease foot contact duration, increase bone mass, and reduce pain. PMID- 15132928 TI - Percutaneous versus open tendo achillis repair. AB - A retrospective analysis was conducted comparing open and percutaneous repair of ruptured Achilles tendon, over a 14-year period. One hundred and eight patients were identified; 70 underwent traditional open repair and 38 had a modified Ma & Griffith repair. The mean operating time with percutaneous repair was 28.5 minutes compared to 45.9 in the open group (p = <.0001). Comparable results were obtained in a subjective analysis of range of movement, stiffness, and power in both groups at time of discharge from clinic. In the open group there were four cases of rerupture (5.7%), four deep infections (4.7%), two palpable suture knots (2.9%), and one sural nerve lesion (1.4%). Complications with percutaneous repair included one rerupture (2.6%), five palpable suture knots (13.2%), four transient sural nerve lesions (10.5%), and no wound infections. There was no statistical significance between the two groups. The authors advocate percutaneous repair, in experienced hands, as a suitable alternative to traditional open techniques. PMID- 15132929 TI - Percutaneous repair of acute Achilles tendon rupture. AB - Various studies have shown that the operative treatment of a freshly ruptured Achilles tendon is generally considered to be more appropriate than a nonoperative regimen. However, complications in open reconstructions are reported to occur in 11-29%. The method used in this study reduced the risk of complications arising from operation, but simultaneously allowed early postoperative mobilization and functional treatment. It was a percutaneous repair of the Achilles tendon, using two Lengemann extension wires for coadaptation of the ruptured tendon. To fix the rupture site, the authors used a fibrin sealant. The spikes of the wire were hooked in at the fascia of the soleus muscle. Via a big, curved needle, the wire was placed in the distal stumps of the ruptured tendon and guided out laterally and medially above the calcaneus. After blocking the wires distally, the fibrin sealant was applied at the rupture site. The current report describes this method of treatment in 66 patients. The postoperative observation period was 1 year. Sixty-four patients were male and two were female. Their average age was 42 years. The Achilles tendon ruptures occurred during sporting activities and were treated by operation within 22 hours on average. The outcome was very good in 98%. One patient (2%) suffered a rerupture due to trauma. There were no other complications. PMID- 15132930 TI - Clinical outcome of tibiotalar arthrodesis utilizing the chevron technique. AB - Tibiotalar arthrodesis remains the gold standard reconstructive procedure for the treatment of disabling ankle arthritis. The purpose of this study was to review the clinical results of tibiotalar arthrodesis utilizing the chevron fusion technique. The results of 46 consecutive patients who underwent ankle arthrodesis utilizing the chevron technique were reviewed. The etiology of the tibiotalar arthritis was posttraumatic in 29 of 46 patients. Of the remaining 17 patients, seven had osteoarthritis, five had talar osteonecrosis, two had rheumatoid arthritis, one had hemophilic arthropathy, one had gouty arthropathy, and one had unrecognized chronic osteomyelitis. Three patients had prior hindfoot arthrodeses, and two patients had bilateral ankle fusions at last follow-up. All patients were followed for a minimum of 2 years. Of the 46 patients, 41 were available for review, with an average follow-up of 7.3 years (range, 2-20 years). Twelve patients had greater than 10-year follow-up. The Mazur ankle score was calculated for all 41 patients. The average Mazur ankle score for the 41 patients available for review was 72.8, out of a maximum possible score of 90. Eighteen patients had excellent results, 11 patients had good results, five patients had fair results, and seven patients had poor results. The most common reasons for fair or poor results were symptomatic subtalar arthritis and multiple medical comorbidities. All patients with postoperative symptomatic subtalar arthritis had preoperative radiographic evidence of subtalar arthrosis. Of the 12 patients with greater than 10-year follow-up, nine had excellent or good results, and an average Mazur ankle score of 76.6. All patients with either prior hindfoot arthrodeses or bilateral ankle fusions had excellent or good results. Of the 41 arthrodeses included in the study, 38 (38/41, 93%) went on to clinical and radiographic union. The chevron technique provides a predictable method to obtain fusion of the tibiotalar joint. Most patients can expect excellent or good results. In the current study, 90% (37/41) of patients were satisfied with the outcome of their surgery and would undergo the same operation again under similar circumstances. PMID- 15132931 TI - Comprehensive reconstruction of the lateral ankle for chronic instability using a free gracilis graft. AB - PURPOSE: The purpose of this retrospective study was to assess the results of a novel surgical technique for the treatment of chronic lateral ankle instability using both a direct repair of the anterior talofibular ligament and a free gracilis tendon transfer to reconstruct anatomically the anterior talofibular and calcaneofibular ligaments. METHODS: Between December 1998 and February 2002, 28 patients (29 ankles) underwent an anatomic reconstruction of the lateral ankle ligaments for chronic ankle instability. Patients returned for a clinical and radiologic follow-up evaluation at an average of 23 months following surgery (range, 12-52 months). Outcomes were assessed by comparison of preoperative and postoperative American Orthopaedic Foot and Ankle Society (AOFAS) scores and visual analog pain scores as well as a postoperative Karlsson score. A subjective self-assessment rating was also obtained. All patients underwent preoperative and postoperative radiographic assessment including talar tilt and anterior drawer stress radiographs. RESULTS: Twenty-eight patients (29 ankles) (100%) returned for final evaluation. Good or excellent outcome was noted on patient subjective self-assessment, pain scores, AOFAS, and Karlsson scores at final follow-up in all patients. Ankle range of motion was not affected by lateral ankle reconstruction. The talar tilt was reduced from a mean of 13 degrees to 3 degrees (p <.0001) and the anterior drawer was reduced from a mean of 10 mm to 5 mm (p <.0001) by the lateral ankle ligamentous reconstruction. CONCLUSION: In the present study, lateral ankle reconstruction with a direct anterior talofibular ligament repair and free gracilis tendon graft augmentation resulted in a high percentage of successful results, excellent ankle stability with a minimal loss of ankle or hindfoot motion, and marked reduction of pain at an average follow-up of almost 2 years. PMID- 15132932 TI - Peroneus brevis is a more effective evertor than peroneus longus. AB - A primary function of the peroneus longus and peroneus brevis is to provide the eversion moment necessary to balance the opposing inversion moments. Surgeons often deal with the loss of or need to sacrifice one of these tendons. This study compares the evertor mechanisms of the peroneus brevis and peroneus longus muscle. This is accomplished in a cadaver model in which the performance of each of the muscle tendons during early heel rise of gait is assessed utilizing the same tendon loads in each so that force is not a variable. Six fresh-frozen cadaver foot-ankle specimens were studied during a simulated early heel rise phase of the gait cycle. The study compared the effect of the peroneus brevis and peroneus longus by separately applying the same load to the each of the tendons. At the talonavicular joint, the peroneus brevis loaded condition externally rotated the navicular 2.1 degrees more than when the peroneus longus was loaded. At the subtalar joint, the peroneus brevis loaded condition resulted in 0.9 degrees more calcaneus valgus relative to the talus than was present during the peroneus longus loaded condition. The experimental data support the hypothesis that the peroneus brevis tendon mechanism is more effective than is the peroneus longus mechanism in rotating the navicular externally and the calcaneus into valgus. This has clinical implications for assisting surgeons in trying to preserve evertor function. PMID- 15132933 TI - Surgical anatomy of the gastrocnemius recession (Strayer procedure). AB - BACKGROUND: The Strayer procedure (gastrocnemius recession) is a treatment option for patients with clinically relevant gastrocnemius equinus contracture. The purpose of this study was to review the surgical anatomy of the Strayer procedure with specific reference to 1) the location of the sural nerve, and 2) the gastrocnemius tendon release point. METHODS: Forty consecutive Strayer procedures in 33 patients (15 males, 18 females) served as the study group. Recorded measurements included: 1) the location of the sural nerve relative to the deep fascia, 2) the distance from the medial border of the gastrocnemius tendon to the sural nerve, and 3) the distance from the distal end of the gastrocnemius muscle belly (identified by surface landmarks) to the actual release site. RESULTS: At the point of the gastrocnemius release, the sural nerve was located superficial to the fascia in 17/40 legs (42.5%) and deep to the fascia in 23/40 legs (57.5%). In five legs (12.5%), the nerve was directly applied to the gastrocnemius tendon and needed to be gently dissected off the tendon. The gastrocnemius release point was located an average of 18 mm distal (range, 20 mm proximal to 57 mm distal) to the surface landmark created by the distal extent of the gastrocnemius muscle belly. CONCLUSION: Knowledge of the relevant anatomy associated with the gastrocnemius recession should allow surgeons to minimize the rate of sural nerve injuries and improve cosmesis by decreasing the length of the surgical incision. A posteromedial incision that begins 2 cm distal to the gastrocnemius indentation and extends proximally will minimize the length of the incision required. PMID- 15132934 TI - Associated deformities and hypermobility in hallux valgus: an investigation with weightbearing radiographs. AB - BACKGROUND: Hallux valgus has been reported to recur after surgical correction in patients subsequently diagnosed with hypermobility of the first ray, pronation of the foot, and pes planovalgus. An objective means of assessing the foot for these deformities preoperatively may avert a poor outcome. This investigation evaluated the efficacy of full-length weightbearing radiographs to recognize associated deformities in patients with hallux valgus before surgery. METHODS: This study compared five parameters from anteroposterior and four parameters from lateral weightbearing pedal radiographs of patients with moderate to severe hallux valgus to a control group to identify differences in the alignment of the midfoot and the first metatarsal-medial cuneiform joint. An examination for clinical evidence of hypermobility was also performed on both groups. RESULTS: The hallux valgus group demonstrated increased abduction and dorsiflexion of the midfoot. The mean talonavicular coverage angle and lateral talo-first metatarsal angle of this group was greater than the mean values for the controls. Radiographic evaluation also revealed differences in the alignment of the first metatarsal-medial cuneiform joint in the sagittal plane. The hallux valgus group possessed a mean of 2 mm of dorsal translation and 2 degrees of dorsiflexion at this joint compared to the controls. CONCLUSIONS: Weightbearing radiographs permit the recognition of associated malalignments of the foot in patients with hallux valgus. The comparisons performed in this study identified deformities consistent with pes planovalgus and hypermobility of the first ray in patients with moderate to severe hallux valgus. The results of our study support a recommendation for a thorough evaluation of full-length, biplanar weightbearing radiographs via the measurement of midfoot and first ray alignment for concomitant deformities of the foot in patients with hallux valgus. PMID- 15132935 TI - Arthrodesis of the toe joints with an intramedullary cannulated screw for correction of hammertoe deformity. AB - Twenty-four patients (37 feet, 51 toes) affected by hammertoe deformity of the lesser toes and treated surgically by arthrodesis of the proximal interphalangeal joint, stabilized with an intramedullary titanium cannulated screw, were reviewed 1-4 years after the operation. At follow-up, the arthrodesis was fused in 48 toes; three toes showed an asymptomatic radiographic nonunion, and in one of them the screw was broken. In seven toes, the cannulated screw was removed because of persistent pain at the tip of the toe where the head of the screw was located. In one case only, there was a late infection, with toe malalignment. All the patients were able to use street shoes 2 weeks after surgery. The average AOFAS score at follow-up was 86.54 points. Compared to the conventional temporary stabilization with an intramedullary Kirschner wire, the stabilization with a cannulated screw decreases the risk of infection, of radiographic nonunion, and of mallet toe deformity. PMID- 15132936 TI - Three-dimensional and two-dimensional computerized tomographic demonstration of calcaneus fractures. AB - The objective of the study was to illustrate the diagnostic quality of three dimensional computed tomography (3DCT) and to document its usefulness for detecting gross changes in articular surfaces of the calcaneus and posterior articular facet (PAF). Superior view 3DCT images of 51 calcanei in 39 different patients were reviewed based on bony fragments produced by primary fracture lines and involvement of the PAF. Of 49 intraarticular fractures examined, there were two severely comminuted and 47 comminuted fractures. Main characteristics of both groups were described. Important findings best seen on 3DCT images were the number and configuration of displaced PAF fragments, fracture lines separating the anterior process and the middle facet, and the extension of fracture lines into the calcaneocuboid facet. The double density sign seen in plain x-rays was identified as a double articular sign in 3DCT images. PMID- 15132937 TI - Intramedullary screw fixation of the fifth metatarsal: an anatomic study and improved technique. AB - Intramedullary screw fixation has been found to be a reliable treatment for certain fractures of the fifth metatarsal. Techniques for this treatment have been described relying largely on intraoperative fluoroscopy. Ten human cadaver specimens had their fifth metatarsals osteotomized and underwent retrograde intramedullary pin placement. Anatomic landmarks and the location of the sural nerve in relation to this starting point were measured. The trajectory of a pin reducing the osteotomy was analyzed. Using the resultant starting point and guide pin trajectory, intramedullary screw placement was performed reliably without the aid of fluoroscopy. This study demonstrates that intramedullary screw fixation of proximal fifth metatarsal fractures may be performed with the use of anatomic landmarks, which decreases the amount of intraoperative fluoroscopy needed. PMID- 15132938 TI - Technique Tip: Modification of DuVries's lesser metatarsophalangeal joint arthroplasty to improve joint mobility. AB - An interposition arthroplasty for the treatment of pain and stiffness in the second metatarsophalangeal joint (MTPJ) is presented. The procedure is a modification of DuVries's lesser MTPJ arthroplasty and is specifically intended to improve limited dorsiflexion not addressed by the original procedure. PMID- 15132939 TI - Responses of ankle musculature of healthy subjects and hemiplegic patients to sinusoidal anterior-posterior movements of the base of support. AB - The responses of the medical gastrocnemius and tibialis anterior muscles to continuous sinusoidal perturbation of the base of support were studied in two groups of subjects, forty-two volunteers aged 20 to 84 years and 17 hemiplegic patients aged 52 to 81 years. EMGs were recorded while subjects stood on a platform oscillating in the anterior-posterior direction. Movement amplitude was adjusted to the maximum the subject could sustain without assisted support within the limits of the instrument. In healthy subjects, two basic activation modes were indicated: (a) a reciprocal contraction pattern in which activity of the medial gastrocnemius and tibialis anterior muscles was concentrated in the anterior and posterior half of the oscillation, respectively. This pattern prevailed in the younger subjects whose maximal movement amplitude reached the highest limit of the instrument; (b) a less discrete pattern, characterized by tonic contraction and periods of coactivation of the two muscles, was seen in older subjects who had difficulties in maintaining stance on the moving platform. The disruption of the discrete reciprocal response mode in this group of patients was related to decline in function of postural mechanisms with aging and demanded additional stiffening of the ankle join, presumably a manifestation of reduced automatism in favor of closer CNS modulation. The typical response mode in the sound leg of the hemiplegic patients was variable coactivation of the two muscles. In the afflicted leg, the establishment of a motor set was impaired: Low tone was associated with negligible muscular activity, although with more elevated tone, low level uniform cocontraction was evident. PMID- 15132940 TI - Skill, strength, handedness, and fatigue. AB - Thirty male subjects, 20 of whom were self-classified left-handers and 10 right handers, were tested on a grip-strength task and a handwriting task with each hand, both under normal conditions and in a situation of induced experimental fatigue. On the basis of questionnaire scores, the left-handers were sub-divided into two groups comprising the 10 most left-handed and the 10 least left-handed subjects. The test of grip strength showed a small but significant deterioration in performance of both hands from fatigue. With the handwriting task, a similar significantly adverse effect of fatigue was recorded for all groups as well as a large significant difference in performance between the preferred and nonpreferred hands under normal conditions, which decreased under fatigue. An explanation of these differential effects is discussed in terms of the greater efficiency of the preferred hand in the highly developed skill of handwriting. PMID- 15132941 TI - A quantitative approach to understanding the formation and change of coordinated movement patterns. AB - This study shows how the identification of relevant collective variables for behavioral patterns and evaluation of their stability may lead to a better understanding of processes underlying pattern change in coordinated human motor behavior. Based upon observed transitions in behavioral patterns, relative phase was identified as an appropriate collective variable for characterizing different patterns of bimanual coordination. The stability of this collective variable was determined through perturbation techniques. Subjects performed one of two patterns of hand coordination (in phase and out of phase) as the frequency of movement was systematically increased. Torque perturbations were administered in order to disturb the ongoing coordinative pattern, and the time required to return to the same pattern (the relaxation time) was measured. A transition from an out-of-phase to an in-phase pattern of coordination occurred at a critical frequency in all subjects. Moreover, when subjects began a trial in the out-of phase pattern, a strong and statistically significant increase in relaxation time was observed as the frequency increased prior to the transition, indicating a loss of pattern stability. No transitions occurred when subjects began in the in phase pattern of coordination; nor was there a change in the measured relaxation time as a function of frequency. Together with earlier results on critical fluctuations, these findings indicate that instabilities play an important role in effecting change in patterns of motor coordination. The potential relevance of these concepts and quantitative tools for other motor activities and other levels of description, for example, neural patterns, is discussed. PMID- 15132942 TI - The appraisal of the velocity-curvature relation in children' hand movements: a research note. AB - A coupling between hand speed and the shape of the hand path appears to be present early in human development. This note addresses the issue of how such a feature may be appraised within digitally recorded trajectories and, in particular, highlights some problems with previously used techniques that may lead to misinterpretation of data. A new approach to such analyses is proposed. PMID- 15132943 TI - Do feedback processing, output variability, and spatial complexity account for manual asymmetries? PMID- 15132944 TI - Manual asymmetries: in defense of a multifactorial account. PMID- 15132945 TI - Report of the 1988 Motor Neuroscience Symposium Collingwood, Ontario, Canada: strange attraction. PMID- 15132946 TI - Recent Advances in Alcoholic Liver Disease I. Role of intestinal permeability and endotoxemia in alcoholic liver disease. AB - A significant body of evidence indicates that endotoxemia and endotoxin-mediated hepatocellular damage play a crucial role in the pathogenesis of alcoholic liver disease. A close correlation between endotoxemia and the severity of alcohol induced liver injury is supported by a number of clinical and experimental studies. Elevated intestinal permeability appears to be the major factor involved in the mechanism of alcoholic endotoxemia and the pathogenesis of alcoholic liver disease. Ethanol and its metabolic derivatives, acetaldehyde in particular, alter intracellular signal-transduction pathways leading to the disruption of epithelial tight junctions and an increase in paracellular permeability to macromolecules. Studies addressing the mechanisms of such epithelial disruption and the protective factors that prevent ethanol and acetaldehyde-mediated disruption of epithelial tight junctions are critically important in the investigations toward the search of preventive and therapeutic strategies for alcoholic liver disease. PMID- 15132947 TI - Gastrointestinal satiety signals IV. Apolipoprotein A-IV. AB - The focus of this article is to review evidence that apolipoprotein A-IV (apo A IV) acts as a satiety factor. Additionally, information regarding the general involvement of apo A-IV in the regulation of food intake and body weight is stated. Apo A-IV is a glycoprotein synthesized by the human intestine. In rodents, both the small intestine and liver secrete apo A-IV, but the small intestine is the major organ responsible for circulating apo A-IV. There is now solid evidence that the hypothalamus, especially the arcuate nucleus, is another active site of apo A-IV expression. Intestinal apo A-IV synthesis is markedly stimulated by fat absorption and does not appear to be mediated by the uptake or reesterification of fatty acids to form triglycerides. Rather, the local formation of chylomicrons acts as a signal for the induction of intestinal apo A IV synthesis. Intestinal apo A-IV synthesis is also enhanced by a factor from the ileum, probably peptide tyrosine-tyrosine (PYY). The inhibition of food intake by apo A-IV is mediated centrally. The stimulation of intestinal synthesis and secretion of apo A-IV by lipid absorption are rapid; thus apo A-IV likely plays a role in the short-term regulation of food intake. Other evidence suggests that apo A-IV may also be involved in the long-term regulation of food intake and body weight, as it is regulated by both leptin and insulin. Chronic ingestion of a high-fat diet blunts the intestinal as well as the hypothalamic apo A-IV response to lipid feeding. It also suppresses apo A-IV gene expression in the hypothalamus. Whereas it is tempting to speculate that apo A-IV may play a role in diet-induced obesity, we believe the confirmation of such a proposal awaits further experimental evidence. PMID- 15132948 TI - G protein-mediated dysfunction of excitation-contraction coupling in ileal inflammation. AB - Inflammation impairs the circular muscle contractile response to muscarinic (M) receptor activation. The aim of this study was to investigate whether the expression of muscarinic receptors, their binding affinity, and the expression and activation of receptor-coupled G proteins contribute to the suppression of contractility in inflammation. The studies were performed on freshly dissociated single smooth muscle cells from normal and inflamed canine ileum. Northern blotting indicated the presence of only M(2) and M(3) receptors on canine ileal circular muscle cells. Inflammation did not alter the mRNA or protein expression of M(2) and M(3) receptors. The maximal binding and K(d) values also did not differ between normal and inflamed cells. However, the contractile response to ACh in M(3) receptor-protected cells was suppressed, whereas that in M(2) receptor-protected cells was enhanced. Further experiments indicated that the expression and binding activity of G alpha(q/11) protein, which couples to M(3) receptors, were downregulated, whereas those of G alpha(i3), which couples to M(2) receptors, were upregulated in inflamed cells. We concluded that inflammation depresses M(3) receptor function, but it enhances M(2) receptor function in ileum. These effects are mediated by the differentially altered expression and binding activity of their respective coupled G alpha(q/11) and G alpha(i3) proteins. PMID- 15132949 TI - Characterization of the rat intestinal Fc receptor (FcRn) promoter: transcriptional regulation of FcRn gene by the Sp family of transcription factors. AB - The regulatory elements that control the transcriptional regulation of the intestinal Fc receptor (FcRn) have not been elucidated. The objective of this study was to characterize the core promoter region of the rat FcRn gene. Chimeric clones that contained various regions of the promoter located upstream of the luciferase reporter were transiently transfected into either IEC-6 or Caco-2 cell lines and nuclear extracts were used to perform DNase I footprint and DNA binding assays (EMSA). Transfection of chimeric upstream nested deletions-luciferase reporter clones into either of these cell lines supported robust reporter activity and identified the location of the minimal promoter at -157/+135. DNase I footprint analysis revealed two complexes located within the gene's core promoter region, and site-directed mutagenesis identified two regions that were critical to maintain basal expression. EMSA identified the presence of five Sp elements within the immediate promoter region that are capable of binding members of the Sp family of proteins. Among the five Sp elements, one element appears to not bind Sp1, Sp2, or Sp3 while influencing the interaction of Sp proteins with an adjacent Sp site. Overexpression of either Sp1 or Sp3 augments activity of the minimal promoter in Sp-deficient Drosophila SL2 cells. In summary, we report on the characterization of the rat FcRn minimal promoter, including the characterization of five Sp elements within this region that interact with members of the Sp family of transcriptional factors and drive promoter activity in intestinal cell lines. PMID- 15132950 TI - Dysregulation of the calpain-calpastatin system plays a role in the development of cerulein-induced acute pancreatitis in the rat. AB - Calpain, a calcium-dependent cytosolic cysteine protease, is implicated in a multitude of cellular functions but also plays a role in cell death. Recently, we have shown that two ubiquitous isoforms, termed micro-calpain and m-calpain, are expressed in rat pancreatic acinar cells and that calcium ionophore-induced calpain activation leads to acinar cell injury. On the basis of these observations, we have now investigated the role of both calpain forms and the endogenous calpain inhibitor calpastatin in acute pancreatitis. After treatment of rats either without or with calpain inhibitor Z-Val-Phe methyl ester (ZVP; 60 mg/kg i.p.), pancreatitis was induced by cerulein injections (10 microg/kg i.p.; 5 times at hourly intervals). Calpain activation and calpastatin expression in the pancreatic tissue were studied by Western blot analysis. Pancreatic injury was assessed by plasma amylase activity, pancreatic wet/dry weight ratio (edema), histological and electron-microscopic analyses, as well as fluorescence labeling of actin filaments. Cerulein caused an activation of both micro-calpain and m calpain, accompanied by degradation of calpastatin. Prophylactic administration of ZVP reduced the cerulein-induced calpain activation but had no effect on calpastatin alterations. In correlation to the diminished calpain activity, the severity of pancreatitis decreased as indicated by a decline in amylase activity (P < 0.01), pancreatic edema formation (P < 0.05), histological score for eight parameters (P < 0.01), and actin filament alterations. Our findings support the hypothesis that dysregulation of the calpain-calpastatin system may play a role in the onset of acute pancreatitis. PMID- 15132951 TI - Medium-chain triglycerides enhance secretory IgA expression in rat intestine after administration of endotoxin. AB - The purpose of this study was to determine whether medium-chain triglycerides (MCTs) modulate the inflammatory immune response to LPS and enhance the expression of secretory IgA in the rat intestine. Rats were given either corn oil or MCTs by gavage daily for 1 wk, and LPS or saline vehicle was administered via the tail vein. They were then killed, and serum and sections from the gut were collected for further analysis. Western blot analysis for secretory IgA revealed that MCTs significantly enhanced its expression in the ileum compared with corn oil in rats administered saline. After LPS challenge, expression of secretory IgA was decreased in the corn oil group but not in the MCTs group. The mRNA expression of IL-6 was assessed by real-time RT-PCR, because IL-6 regulates secretory IgA in the intestine. The expression was significantly greater in the MCTs group than in the corn oil group after LPS injection. Increases in expression of proinflammatory cytokines or chemokines such as TNF-alpha, IL-18, macrophage inflammatory protein-2, and monocyte chemoattractant protein-1 in the ileum were significantly blunted by MCTs. In addition, the mRNA expression of the Th2 IgA-stimulating cytokine IL-10 in the ileum and Peyer's patches was significantly greater in the MCTs than the corn oil group. In contrast, the mRNA expression of the Th1 IgA-inhibiting cytokine interferon-gamma was blunted by MCTs. As a result, intestinal injury was significantly reduced. Therefore, MCTs protect the gut by modulating the immune response to LPS and enhancing secretory IgA expression. PMID- 15132952 TI - Smad and p38-MAPK signaling mediates apoptotic effects of transforming growth factor-beta1 in human airway epithelial cells. AB - Transforming growth factor-beta1 (TGF-beta1) belongs to a family of multifunctional cytokines that regulate a variety of biological processes, including cell differentiation, proliferation, and apoptosis. The effects of TGF beta1 are cell context and cell cycle specific and may be signaled through several pathways. We examined the effect of TGF-beta1 on apoptosis of primary human central airway epithelial cells and cell lines. TGF-beta1 protected human airway epithelial cells from apoptosis induced by either activation of the Fas death receptor (CD95) or by corticosteroids. This protective effect was blocked by inhibition of the Smad pathway via overexpression of inhibitory Smad7. The protective effect is associated with an increase in the cyclin-dependent kinase inhibitor p21 and was blocked by the overexpression of key gatekeeper cyclins for the G1/S interface, cyclins D1 and E. Blockade of the Smad pathway by overexpression of the inhibitory Smad7 permitted demonstration of a TGF-beta mediated proapoptotic pathway. This proapoptotic effect was blocked by inhibition of the p38 MAPK kinase signaling with the inhibitor SB-203580 and was associated with an increase in p38 activity as measured by a kinase assay. Here we demonstrate dual signaling pathways involving TGF-beta1, an antiapoptotic pathway mediated by the Smad pathway involving p21, and an apoptosis-permissive pathway mediated in part by p38 MAPK. PMID- 15132953 TI - Greater vascularity, lowered HIF-1/DNA binding, and elevated GSH as markers of adaptation to in vivo chronic hypoxia. AB - Vascularity is increased in placentas from high- compared with low-altitude pregnancies. An angiogenic response to hypoxia may protect an organ from further hypoxic insult by increasing blood flow and oxygen delivery to the tissue. We hypothesized that increased placental vascularity is sufficient to adapt to high altitude. Therefore, indexes of hypoxic stress would not be present in placentas from successful high-altitude pregnancies. Full-thickness placental biopsies were 1) collected and frozen in liquid nitrogen within 5 min of placental delivery and 2) fixed in formalin for stereologic analyses at high (3,100 m, n = 10) and low (1,600 m, n = 10) altitude. Hypoxia-inducible transcription factor (HIF-1) activity was analyzed by ELISA. Western blot analyses were used to evaluate HIF 1alpha, HIF-1beta, HIF-2alpha, von Hippel-Lindau protein, VEGF, Flt-1, enolase, and GAPDH. Magnetic resonance spectroscopy was used to evaluate endogenous metabolism. The ratio of placental capillary surface density to villous surface density was 70% greater at high compared with low altitude. HIF-1 activity and HIF-1-associated proteins were unchanged in placentas from high- vs. low-altitude pregnancies. Placental expression of HIF-1-mediated proteins VEGF, Flt-1, enolase, and GAPDH were unchanged at high vs. low altitude. Succinate, GSH, phosphomonoesters, and ADP were elevated in placenta from high compared with low altitude. Placentas from uncomplicated high-altitude pregnancies have greater vascularity and no indication of significant hypoxic stress at term compared with placentas from low altitude. PMID- 15132954 TI - Prevention and reversal of pulmonary inflammation and airway hyperresponsiveness by dexamethasone treatment in a murine model of asthma induced by house dust. AB - The morbidity and mortality from asthma in the Western world have increased 75% in the past 20 years. Recent studies have demonstrated that sensitization to cockroach allergens correlates strongly with the increased asthma morbidity for adults and children. We investigated whether dexamethasone administered before or after allergen challenge would inhibit the pulmonary inflammation and airway hyperresponsiveness in a mouse model of asthma induced by a house dust extract with high levels of cockroach allergens. For the prevention experiment, mice were treated with an intraperitoneal injection of dexamethasone 1 h before each pulmonary challenge, and airway hyperresponsiveness was measured 24 h after the last challenge. Mice were killed 48 h after the last challenge. For the reversal study, airway hyperresponsiveness was measured 24 h after the last challenge, and the mice were treated with dexamethasone. Dexamethasone treatment before allergen challenge significantly reduced the pulmonary recruitment of inflammatory cells, myeloperoxidase activity in the lung, airway hyperreactivity, and total serum IgE levels compared with PBS-treated mice. Additionally, dexamethasone treatment could significantly reduce the airway hyperreactivity of an established asthmatic response. These results demonstrate that dexamethasone not only prevents but also halts the asthmatic response induced by house dust containing cockroach allergens. This model exhibits several features of human asthma that may be exploited in the study of pathophysiological mechanisms and potential therapeutic interventions. PMID- 15132955 TI - Airway obstruction and stenting: more complex than we thought? PMID- 15132956 TI - Treatment of multidrug-resistant tuberculosis: should you try this at home? PMID- 15132957 TI - American in name, international in scope. PMID- 15132958 TI - Mycobacterium tuberculosis and the cause of consumption: from discovery to fact. PMID- 15132959 TI - Stenting at the flow-limiting segment in tracheobronchial stenosis due to lung cancer. AB - Airway stenting at the wave-speed flow-limiting segment (the choke point) is assessed. We determined prospectively the precise location of the choke point using the flow-volume curve, endobronchial ultrasonography, ultrathin bronchoscopy, and three-dimensional computed tomography scan before and after stenting in 64 patients with extrincic compression due to lung cancer. We noted distinct flow-volume curve patterns specific to the type of stenosis. The tracheal stenosis group indicated fixed narrowing patterns with an expiratory plateau, bronchial stenosis group dynamic collapse patterns with an expiratory flow deterioration (choking), carinal stenosis group combined fixed and dynamic patterns, and extensive stenosis group complex patterns containing elements of all the former. After stenting, almost full-function patterns with significant improvement in PEF were observed in all groups (p < 0.01, p < 0.05, p < 0.001, p < 0.01, respectively). In patients with extensive stenosis, implantation of additional stents was required when the choke points were observed to have migrated to the areas of malacia with cartilage destruction by the tumor. Secondary stenting at migrated choke points resulted in a significant improvement in PEF over the initial stenting (p < 0.01). Stenting at the choke point improved expiratory flow limitation by increasing the cross-sectional area, supporting the weakened airway wall and relieving dyspnea. PMID- 15132960 TI - Executive summary on the systematic review and practice parameters for portable monitoring in the investigation of suspected sleep apnea in adults. PMID- 15132961 TI - "Failure to communicate" due to forgetting "marriage vow"? PMID- 15132962 TI - Laennec, trema, and tuberculosis. PMID- 15132963 TI - Inhaled corticosteroids in COPD and mortality: inaccuracies? PMID- 15132964 TI - Antimicrobial treatment of ventilator-associated pneumonia. PMID- 15132965 TI - Type 2 cytokines in respiratory syncytial virus bronchiolitis. PMID- 15132966 TI - Side effects of antituberculosis therapy. PMID- 15132967 TI - Design-based counting. PMID- 15132968 TI - Hedgehog signaling promotes prostate xenograft tumor growth. AB - During fetal prostate development, Sonic hedgehog (Shh) expression by the urogenital sinus epithelium activates Gli-1 expression in the adjacent mesenchyme and promotes outgrowth of the nascent ducts. Shh signaling is down-regulated at the conclusion of prostate ductal development. However, a survey of adult human prostate tissues reveals substantial levels of Shh signaling in normal, hyperplasic, and malignant prostate tissue. In cancer specimens, the Shh expression is localized to the tumor epithelium, whereas Gli-1 expression is localized to the tumor stroma. Tight correlation between the levels of Shh and Gli-1 expression suggests active signaling between the tissue layers. To determine whether Shh-Gli-1 signaling could be functionally important for tumor growth and progression, we performed experiments with the LNCaP xenograft tumor model and demonstrated that: 1). Shh expressed by LNCaP tumor cells activates Gli 1 expression in the tumor stroma, 2). genetically engineered Shh overexpression in LNCaP cells leads to increased tumor stromal Gli-1 expression, and 3). Shh overexpression dramatically accelerates tumor growth. These data suggest that hedgehog signaling from prostate cancer cells to the stroma can elicit the expression of paracrine signals, which promote tumor growth. PMID- 15132969 TI - Energy partitioning in gluteal-femoral fat: does the metabolic fate of triglycerides affect coronary heart disease risk? PMID- 15132970 TI - PPAR-gamma agonists: shifting attention from the belly to the heart? PMID- 15132971 TI - Protective role of imatinib in atherosclerosis. PMID- 15132972 TI - Control of smooth muscle cell activation. PMID- 15132973 TI - Ceramide synthesis correlates with the posttranscriptional regulation of the sterol-regulatory element-binding protein. AB - OBJECTIVE: Sterol-regulatory element-binding proteins (SREBPs) regulate transcription of genes of lipid metabolism. Ceramide decreases transcriptionally active SREBP levels independently of intracellular cholesterol levels. Mechanisms of the ceramide-mediated decrease of SREBP levels were investigated. METHODS AND RESULTS: Experiments were performed in Chinese hamster ovary cells. Inhibition of ceramide synthesis with myriocin, cycloserine, or fumonisin decreases levels of transcriptionally active SREBP and reduces SRE-mediated gene transcription. When ceramide synthesis is increased through exogenous sphingosine or inhibition of sphingosine kinase, SRE-mediated gene transcription is increased. The important role of ceramide synthesis in SRE-mediated gene transcription is confirmed in LY B cells that do not synthesize ceramide de novo. LY-B cells fail to increase SRE mediated gene transcription in sterol depletion. CONCLUSIONS: Ceramide synthesis correlates with the generation of transcriptionally active SREBP and SRE-mediated gene transcription. Inhibition of ceramide synthesis decreases levels of transcriptionally active SREBP and SRE-mediated gene transcription. It is hypothesized that the process of ongoing ceramide synthesis contributes to the physiological processing of SREBP, perhaps affecting ER-to-Golgi trafficking. Taken together, modification of ceramide synthesis could be a novel target for drug development in the pharmacologic modification of SRE-dependent pathways. PMID- 15132974 TI - Carbon monoxide improves cardiac energetics and safeguards the heart during reperfusion after cardiopulmonary bypass in pigs. AB - Ischemia-reperfusion injury, a clinical problem during cardiac surgery, involves worsened adenosine trisphosphate (ATP) generation and damage to the heart. We studied carbon monoxide (CO) pretreatment, proven valuable in rodents but not previously tested in large animals, for its effects on pig hearts subjected to cardiopulmonary bypass with cardioplegic arrest. Hearts of CO-treated pigs showed significantly higher ATP and phosphocreatine levels, less interstitial edema, and apoptosis of cardiomyocytes and required fewer defibrillations after bypass. We conclude that treatment with CO improves the energy status, prevents edema formation and apoptosis, and facilitates recovery in a clinically relevant model of cardiopulmonary bypass surgery. PMID- 15132975 TI - Estrogen and neuroprotection: higher constitutive expression of glutaredoxin in female mice offers protection against MPTP-mediated neurodegeneration. AB - Incidence of Parkinson's disease is lower in women as compared with men. Although neuroprotective effect of estrogen is recognized, the underlying molecular mechanisms are unclear. MPTP (1-methyl-4-phenyl-1, 2, 3, 6, tetrahydro-pyridine), a neurotoxin that causes Parkinson's disease-like symptoms acts through inhibition of mitochondrial complex I. Administration of MPTP to male mice results in loss of dopaminergic neurons in substantia nigra, whereas female mice are unaffected. Oxidation of critical thiol groups by MPTP disrupts mitochondrial complex I, and up-regulation of glutaredoxin (a thiol disulfide oxidoreductase) is essential for recovery of complex I. Early events following MPTP exposure, such as increased AP1 transcription, loss of glutathione, and up-regulation of glutaredoxin mRNA is seen only in male mice, indicating that early response to neurotoxic insult does not occur in females. Pretreatment of female mice with ICI 182,780, estrogen receptor (ER) antagonist sensitizes them to MPTP-mediated complex I dysfunction. Constitutive expression of glutaredoxin is significantly higher in female mice as compared with males. ICI 182,780 down-regulates glutaredoxin activity in female mouse brain regions (midbrain and striatum), indicating that glutaredoxin expression is regulated through estrogen receptor signaling. Higher constitutive expression of glutaredoxin could potentially contribute to the neuroprotection seen in female mouse following exposure to neurotoxins, such as MPTP. PMID- 15132976 TI - An essential role of Cav1.2 L-type calcium channel for urinary bladder function. AB - Mice deficient in the smooth muscle Cav1.2 calcium channel (SMACKO, smooth muscle alpha1c-subunit calcium channel knockout) have a severely reduced micturition and an increased bladder mass. L-type calcium current, protein, and spontaneous contractile activity were absent in the bladder of SMACKO mice. K+ and carbachol (CCh)-induced contractions were reduced to 10-fold in detrusor muscles from SMACKO mice. The dihydropyridine isradipine inhibited K+- and CCh-induced contractions of muscles from CTR but had no effect in muscles from SMACKO mice. CCh-induced contraction was blocked by removing extracellular Ca2+ but was unaffected by the PLC inhibitor U73122 or depletion of intracellular Ca2+ stores by thapsigargin. In muscles from CTR and SMACKO mice, CCh-induced contraction was partially inhibited by the Rho-kinase inhibitor Y27632. These results show that the Cav1.2 Ca2+ channel is essential for normal bladder function. The Rho-kinase and Ca2+-release pathways cannot compensate the lack of the L-type Ca2+ channel. PMID- 15132977 TI - Triacylglycerol accumulation in human obesity and type 2 diabetes is associated with increased rates of skeletal muscle fatty acid transport and increased sarcolemmal FAT/CD36. AB - We examined whether, in human obesity and type 2 diabetes, long chain fatty acid (LCFA) transport into skeletal muscle is upregulated and contributes to an excess intramuscular triacylglycerol accumulation. In giant sarcolemmal vesicles prepared from human skeletal muscle, LCFA transport rates were upregulated approximately 4-fold and were associated with an increased intramuscular triacylglycerol content in obese individuals and in type 2 diabetics. In these individuals, the increased sarcolemmal LCFA transport rate was not associated with an altered expression of FAT/CD36 or FABPpm. Instead, the increase in the LCFA transport rate was associated with an increase in sarcolemmal FAT/CD36 but not sarcolemmal FABPpm. Rates of fatty acid esterification were increased threefold in isolated human muscle strips obtained from obese subjects, while concomitantly rates of fatty acid oxidation were not altered. Thus, the increased rate of fatty acid transport may contribute to the increased rates of triacylglycerol accumulation in human skeletal muscle. The altered FAT/CD36 trafficking in muscle from obese subjects and type 2 diabetics juxtaposes the known alterations in GLUT4 trafficking, i.e., GLUT4 is known to be retained in its intracellular depots while FAT/CD36 is retained at the sarcolemma. This redistribution of FAT/CD36 to the sarcolemma may contribute to the etiology of insulin resistance in human muscle, and hence, FAT/CD36 provides another potential therapeutic target for the prevention and/or treatment of insulin resistance. PMID- 15132978 TI - Structurally different RGTAs modulate collagen-type expression by cultured aortic smooth muscle cells via different pathways involving fibroblast growth factor-2 or transforming growth factor-beta1. AB - We have engineered polymers called ReGeneraTing Agents (RGTAs), which mimic the protecting and potentiating properties of heparan sulfates toward heparin-binding growth factors (HBGF). RGTAs have been shown to optimize cell growth and regulate collagen production in vitro. Here, we studied relationships between RGTA structure and collagen-type expression in aortic smooth muscle cells by using two RGTAs, the carboxylmethylsulfate dextran RG-1503 and the carboxylmethylsulfate dextran with added benzylamide RG-1192. RG-1192 specifically induced a fivefold decrease in collagen III synthesis. This effect was abolished by FGF-2 neutralizing antibody. RG-1192 and FGF-2 acted synergistically to decrease collagen III. RG-1192 was more effective than heparin in this process. RG-1192 increased the pericellular localization of FGF-2 and protected FGF-2 from proteolysis. Surface plasmon resonance analysis indicated a Kd of 15.7 nM for the RG-1192/FGF-2 interaction (10.6 nM for the heparin/FGF-2 interaction). The structurally different RG-1503 (without benzylamide) did not interact with FGF-2 and worked synergistically with TGF-beta1 to specifically induce a twofold increase in collagen V. RGTAs with different structures exert different modulating effects on the collagen phenotype. Selection of appropriate RGTAs, which had been shown to enhance in vivo tissue repair, may provide a mean of correcting collagen abnormalities in vascular disorders and more generally in fibrotic diseases. PMID- 15132979 TI - Hypoxia induces macrophage inflammatory protein-2 (MIP-2) gene expression in murine macrophages via NF-kappaB: the prominent role of p42/ p44 and PI3 kinase pathways. AB - We have previously reported that hypoxia induces a pronounced inflammatory response in the mouse lung associated with elevated levels of specific chemokines. To further explore the mechanisms involved in lung inflammation, we exposed RAW 264.7 cells as well as mouse primary macrophages to hypoxia and analyzed chemokine gene expression. Among the genes examined, macrophage inflammatory protein-2 (MIP-2) expression was prominently induced by hypoxia both at the mRNA and the protein level. When RAW 264.7 cells were transfected with a panel of plasmids harboring a luciferase marker gene under the control of wild type or mutant variants of the MIP-2 gene promoter, a strong hypoxic induction of expression (9- to 17-fold) was observed. This induction was abolished by a mutation targeted to an NF-kappaB binding site in the MIP-2 promoter. Concordantly, specific NF-kappaB binding to the cognate sequence was enriched in nuclear extracts from hypoxic but not normoxic RAW 264.7 cells. The mechanism of MIP-2 gene induction by hypoxia was further characterized using inhibitors of signaling kinases. Inhibition of the p42/p44 and PI3 kinases but not p38 MAPK abolished the NF-kappaB-driven upregulation of MIP-2 gene expression by hypoxia. This attenuation of the NF-kappaB response to hypoxia did not involve decreased nuclear NF-kappaB abundance but correlated with diminished transactivation potential of the p65 subunit. Our results indicate that the hypoxic signal for induction of MIP-2 gene expression is implemented through enhanced NF-kappaB activity and transmitted along the p42/44 and PI3 kinase pathways. PMID- 15132980 TI - Cardiac myocyte-specific HIF-1alpha deletion alters vascularization, energy availability, calcium flux, and contractility in the normoxic heart. AB - At a resting pulse rate the heart consumes almost twice-as much oxygen per gram tissue as the brain and more than 43 times more than resting skeletal muscle (1). Unlike skeletal muscle, cardiac muscle cannot sustain anaerobic metabolism. Balancing oxygen demand with availability is crucial to cardiac function and survival, and regulated gene expression is a critical element of maintaining this balance. We investigated the role of the hypoxia-inducible transcription factor HIF-1alpha in maintaining this balance under normoxic conditions. Cardiac myocyte specific HIF-1alpha gene deletion in the hearts of genetically engineered mice caused reductions in contractility, vascularization, high-energy phosphate content, and lactate production. This was accompanied by altered calcium flux and altered expression of genes involved in calcium handling, angiogenesis, and glucose metabolism. These findings support a central role for HIF-1alpha in coordinating energy availability and utilization in the heart and have implications for disease states in which cardiac oxygen delivery is impaired. Heart muscle requires a constant supply of oxygen. When oxygen supply does not match myocardial demand cardiac contractile dysfunction occurs, and prolongation of this mismatch leads to apoptosis and necrosis. Coordination of oxygen supply and myocardial demand involves immediate adaptations, such as coronary vasodilatation, and longer-term adaptations that include altered patterns of gene expression (2-4). How the expression of multiple genes is coordinated with oxygen availability in the heart and the impact of oxygen-dependent gene expression on cardiac function are insufficiently understood. Further elucidating these relationships may help clarify the molecular pathology of various cardiovascular disease states, including ischemic cardiomyopathy and myocardial hibernation (5, 6). PMID- 15132981 TI - Role of myoglobin in the antioxidant defense of the heart. AB - Although the primary function of myoglobin (Mb) has been considered to be cellular O2 storage and supply, recent studies have shown that Mb in addition can act as NO oxidase. Here we report that Mb also significantly contributes to the attenuation of oxidative stress in cardiac muscle. In support of this hypothesis, we found that in isolated perfused hearts of Mb-deficient (myo-/-) mice oxidative challenge by intracoronary infused H2O2 (1-300 microM) or superoxide formed by 2,3-dimethoxy-1,4-naphtoquinone (0.1-30 microM), respectively, depressed cardiac contractility to a greater extent than in wild-type (WT) hearts, e.g., up to [H2O2] = 10 microM there was a significant left ventricular developed pressure (LVDP) decrease in myo-/- hearts only (90.4+/-4.2 vs. 98.1+/-0.7% of control, n=6, P<0.05). Likewise in an ischemia/reperfusion protocol, myo-/- hearts showed a delayed recovery of postischemic function as compared with WT controls (e.g., LVDP was 35.6+/-7.5 vs. 22.4+/-5.3 mmHg, respectively, after 10 min of reperfusion, P<0.05, n=8), which correlated well with an enhanced release of reactive oxygen species in myo-/- hearts as measured by online lucigenin-enhanced chemiluminescence [e.g. 465+/-87 relative light units (RLU) in myo-/- vs. 287+/ 73 RLU in WT after 2.5 min of reperfusion, P<0.05, n=8]. (31)P NMR spectroscopy revealed concomitantly a more pronounced phosphocreatine overshoot during reperfusion in the knockout but only minute alterations in ATP and pHi. Our data show that lack of Mb leads to increased vulnerability of cardiac function to oxidative challenge either pharmacologically induced or endogenously generated. We propose that Mb is a key element influencing redox pathways in cardiac muscle to functionally and metabolically protect the heart from oxidative damage. PMID- 15132982 TI - Apoptotic adaptations from exercise training in skeletal and cardiac muscles. AB - The effect of exercise on apoptosis in postmitotic tissues is not known. In this study, we investigated the effect of regular moderate physical activity (i.e., exercise training) on the extent of apoptosis in rat skeletal and cardiac muscles. Adult Sprague Dawley rats were trained (TR) 5 days weekly for 8 wk on treadmill. Sedentary rats served as controls (CON). An ELISA was used to detect mono- and oligonucleosome fragmentation as an indicator of apoptosis. Bcl-2, Bax, Apaf-1, AIF, cleaved PARP, cleaved caspase-3, cleaved/active caspase-9, heat shock protein (HSP)70, Cu/Zn-SOD, and Mn-SOD protein levels were determined by Western analyses. Bcl-2 and Bax transcript contents were estimated by RT-PCR. A spectrofluorometric assay was used to determine caspase-3 activity. DNA fragmentation in ventricles of the TR group decreased by 15% whereas that in soleus of the TR group tended to decrease (P=0.058) when compared with CON group. Protein contents of Bcl-2, HSP70, and Mn-SOD increased in both soleus and ventricle muscles of TR animals when compared with CON animals. Apaf-1 protein content in the soleus of TR animals was lower than that of CON animals. Bcl-2 mRNA levels increased in both ventricle and soleus muscles of TR animals, and Bax mRNA levels decreased in the soleus of TR animals when compared with CON animals. Furthermore, HSP70 protein content was negatively correlated to Bax mRNA content and was positively correlated to Bcl-2 protein and mRNA contents. Mn-SOD protein content was negatively correlated to the apoptotic index, and caspase-3 activity and was positively correlated to Bcl-2 transcript content and HSP70 protein content. These data suggest that exercise training attenuates the extent of apoptosis in cardiac and skeletal muscles. PMID- 15132983 TI - Nitric oxide levels regulate macrophage commitment to apoptosis or necrosis during pneumococcal infection. AB - Macrophages are resistant to constitutive apoptosis, but infectious stimuli can induce either microbial or host-mediated macrophage apoptosis. Phagocytosis and killing of opsonized pneumococci by macrophages are potent stimuli for host mediated apoptosis, but the link between pneumococcal killing and apoptosis induction remains undefined. We now show phagocytosis of pneumococci by differentiated human monocyte-derived macrophages (MDM) results in up-regulation of inducible nitric oxide synthase (iNOS) and increased production of NO and reactive nitrogen species. NO accumulation in macrophages initiates an apoptotic program that involves NO-dependent mitochondrial membrane permeabilization, Mcl-1 down-regulation, and caspase activation and results in nuclear condensation and fragmentation. An inhibitor of mitochondrial permeability transition, bongkrekic acid, decreases pneumococcal-associated macrophage apoptosis. Conversely, inhibition of NO production using iNOS inhibitors decreases bacterial killing and shifts the cell death program from apoptosis to necrosis. Pneumolysin contributes to both NO production and apoptosis induction. After initial microbial killing, NO accumulation switches the macrophage phenotype from an activated cell to a cell susceptible to apoptosis. These results illustrate important roles for NO in the integration of host defense and regulation of inflammation in human macrophages. PMID- 15132984 TI - Cross-linking of ubiquitin, HSP27, parkin, and alpha-synuclein by gamma-glutamyl epsilon-lysine bonds in Alzheimer's neurofibrillary tangles. AB - The accumulation of misfolded proteins in intracellular inclusions is a generic feature of neurodegenerative disorders. Although heavily ubiquitylated, the aggregated proteins are not degraded by the proteasomes. A possible reason for this phenomenon may be a modification of deposited proteins by transglutaminases forming gamma-glutamyl-epsilon-lysine (GGEL) cross-links between distinct proteins. Here, we show that the frequency of GGEL cross-links is an order of magnitude higher in Alzheimer's brain cortex than in age-matched or younger controls. This difference is due to the accumulation of GGEL cross-links in ubiquitin-immunopositive protein particles present in both Alzheimer's brains and those from aged individuals. The highly cross-linked protein aggregates show immunoreactivity to antibodies against tau and neurofilament proteins, and partially also to alpha-synuclein, indicating that these structures are inherent in Alzheimer's neurofibrillary tangles and Lewy bodies. Using mass sequence analysis, we identified the same six pairs of peptide sequences cross-linked in both senile and Alzheimer's specimens: Gln31 and Gln190 of HSP27 protein are cross-linked with Lys29 and Lys48 of ubiquitin and HSP27 therefore may cross-link two (poly)ubiquitin chains. One lysine residue of parkin and one of alpha synuclein were also found to be cross-linked. The data suggest that cross-linking of (poly)ubiquitin moieties via HSP27 may have a role in the stabilization of the intraneuronal protein aggregates by interference with the proteasomal elimination of unfolded proteins. PMID- 15132985 TI - Nitric oxide, a key signaling molecule in the murine early embryonic heart. AB - Nitric oxide (NO) is thought to play an important role as a signaling molecule in embryonic and adult cardiomyocytes; however, its involvement in muscarinic signaling is still unclear. The aim of the present work was to analyze the muscarinic modulation of the L-type Ca2+ current (ICa) in early- and late-stage embryonic ventricular cardiomyocytes. Muscarinic stimulation depressed basal ICa by 30.1 +/- 3.2% (n=27) in early-stage cardiomyocytes. Pharmacological evidence suggested that the muscarinic modulation was mediated through generation of NO, activation of cGMP-dependent phosphodiesterase (PDE) 2, and ensuing lowering of cyclic AMP/protein kinase A (cAMP/PKA) levels. Conversely, in late-stage cardiomyocytes, muscarinic regulation of ICa occurred in a NO-independent manner via inhibition of prestimulated adenylyl cyclase (AC). To unequivocally prove the involvement of NO and to identify the nitric oxide synthase (NOS) isoform(s), we analyzed muscarinic signaling in embryonic ventricular cardiomyocytes of NOS2 (-/ ) and NOS3 (-/-) mice. The early-stage NOS3 (-/-) cardiomyocytes lacked muscarinic modulation, whereas it was preserved in NOS2 (-/-) cells. Moreover, at the late embryonic stage, muscarinic modulation of ICa was intact in both strains. Thus, NO is the key regulator of muscarinic signaling in the early embryonic ventricle, whereas at later stages, signaling occurs through a NO independent pathway. PMID- 15132986 TI - Neuroprotective effects of spermine following hypoxic-ischemic-induced brain damage: a mechanistic study. AB - The polyamines (spermine, putrescine, and spermidine) can have neurotoxic or neuroprotective properties in models of neurodegeneration. However, assessment in a model of hypoxia-ischemia (HI) has not been defined. Furthermore, the putative mechanisms of neuroprotection have not been elucidated. Therefore, the present study examined the effects of the polyamines in a rat pup model of HI and determined effects on key enzymes involved in inflammation, namely, nitric oxide synthase (NOS) and arginase. In addition, effects on mitochondrial function were investigated. The polyamines or saline were administered i.p. at 10mg/kg/day for 6 days post-HI. Histological assessment 7 days post-HI revealed that only spermine significantly (P<0.01) reduced infarct size from 46.14 +/- 10.4 mm3 (HI + saline) to 4.9 +/- 2.7 mm3. NOS activity was significantly increased following spermine treatment in the left (ligated) hemisphere compared with nonintervention controls (P<0.01) and HI + saline (P<0.05). In contrast, spermine decreased arginase activity compared with HI + saline but was still significantly elevated in comparison to nonintervention controls (P<0.01). Assessment of mitochondrial function in the HI + saline group, revealed significant and extensive damage to complex-I (P<0.01) and IV (P<0.001) and loss of citrate synthase activity (P<0.05). No effect on complex II-III was observed. Spermine treatment significantly prevented all these effects. This study has therefore confirmed the neuroprotective effects of spermine in vivo. However, for the first time, we have shown that this effect may, in part, be due to increased NOS activity and preservation of mitochondrial function. PMID- 15132987 TI - Glycogen synthase kinase 3beta (GSK3beta) mediates 6-hydroxydopamine-induced neuronal death. AB - The causes of sporadic Parkinson's disease (PD) are poorly understood. 6 Hydroxydopamine (6-OHDA), a PD mimetic, is widely used to model this neurodegenerative disorder in vitro and in vivo; however, the underlying mechanisms remain incompletely elucidated. We demonstrate here that 6-OHDA evoked endoplasmic reticulum (ER) stress, which was characterized by an up-regulation in the expression of GRP78 and GADD153 (Chop), cleavage of procaspase-12, and phosphorylation of eukaryotic initiation factor-2 alpha in a human dopaminergic neuronal cell line (SH-SY5Y) and cultured rat cerebellar granule neurons (CGNs). Glycogen synthase kinase-3 beta (GSK3beta) responds to ER stress, and its activity is regulated by phosphorylation. 6-OHDA significantly inhibited phosphorylation of GSK3beta at Ser9, whereas it induced hyperphosphorylation of Tyr216 with little effect on GSK3beta expression in SH-SY5Y cells and PC12 cells (a rat dopamine cell line), as well as CGNs. Furthermore, 6-OHDA decreased the expression of cyclin D1, a substrate of GSK3beta, and dephosphorylated Akt, the upstream signaling component of GSK3beta. Protein phosphatase 2A (PP2A), an ER stress-responsive phosphatase, was involved in 6-OHDA-induced GSK3beta dephosphorylation (Ser9). Blocking GSK3beta activity by selective inhibitors (lithium, TDZD-8, and L803-mts) prevented 6-OHDA-induced cleavage of caspase-3 and poly(ADP-ribose) polymerase (PARP), DNA fragmentations and cell death. With a tetracycline (Tet)-controlled TrkB inducible system, we demonstrated that activation of TrkB in SH-SY5Y cells alleviated 6-OHDA-induced GSK3beta dephosphorylation (Ser9) and ameliorated 6-OHDA neurotoxicity. TrkB activation also protected CGNs against 6-OHDA-induced damage. Although antioxidants also offered neuroprotection, they had little effect on 6-OHDA-induced GSK3beta activation. These results suggest that GSK3beta is a critical intermediate in pro apoptotic signaling cascades that are associated with neurodegenerative diseases, thus providing a potential target site amenable to pharmacological intervention. PMID- 15132988 TI - Toll-like receptor 4 functions intracellularly in human coronary artery endothelial cells: roles of LBP and sCD14 in mediating LPS responses. AB - Endothelial cells are activated by microbial agonists through Toll-like receptors (TLRs) to express inflammatory mediators; this is of significance in acute as well as chronic inflammatory states such as septic shock and atherosclerosis, respectively. We investigated mechanisms of lipopolysaccharide (LPS)-induced cell activation in human coronary artery endothelial cells (HCAEC) using a combination of FACS, confocal microscopy, RT-PCR, and functional assays. We found that TLR4, in contrast to TLR2, is not only located intracellularly but also functions intracellularly. That being the case, internalization of LPS is required for activation. We further characterized the HCAEC LPS uptake system and found that HCAEC exhibit an effective LPS uptake only in the presence of LPS binding protein (LBP). In addition to its function as a catalyst for LPS-CD14 complex formation, LBP enables HCAEC activation at low LPS concentrations by facilitating the uptake, and therefore delivery, of LPS-CD14 complexes to intracellular TLR4-MD-2. LBP-dependent uptake involves a scavenger receptor pathway. Our findings may be of pathophysiological relevance in the initial response of the organism to infection. Results further suggest that LBP levels, which vary as LBP is an acute phase reactant, could be relevant to initiating inflammatory responses in the vasculature in response to chronic or recurring low LPS. PMID- 15132989 TI - Molecular imaging of homodimeric protein-protein interactions in living subjects. AB - Homodimeric protein interactions are potent regulators of cellular functions, but are particularly challenging to study in vivo. We used a split synthetic renilla luciferase (hRLUC) complementation-based bioluminescence assay to study homodimerization of herpes simplex virus type 1 thymidine kinase (TK) in mammalian cells and in living mice. We quantified and imaged homodimerization of TK chimeras containing N-terminal (N-hRLUC) or C-terminal (C-hRLUC) fragments of hRLUC in the upstream and downstream positions, respectively (tail-to-head homodimer). This was monitored using luminometry (68-fold increase, and was significantly [P<0.01] above background light emission) and by CCD camera imaging of living mice implanted with ex vivo transfected 293T cells (2.7-fold increase, and is significantly [P<0.01] above background light emission). We also made a mutant-TK to generate N-hRLUC mutant TK and mutant TK-C-hRLUC by changing a single amino acid at position 318 from arginine to cysteine, a key site that has previously been reported to be essential for TK homo-dimerization, to support the specificity of the hRLUC complementation signal from TK homodimerization. Ex vivo substrate (8-3H Penciclovir) accumulation assays in 293T cells expressing the TK protein chimeras showed active TK enzyme. We also devised an experimental strategy by constructing variant TK chimeras (possessing extra N-hRLUC or C-hRLUC 'spacers') to monitor incremental lack of association of the tail-to-head TK homodimer. Application of this potentially generalizable assay to screen for molecules that promote or disrupt ubiquitous homodimeric protein-protein interactions could serve not only as an invaluable tool to understand biological networks but could also be applied to drug discovery and validation in living subjects. PMID- 15132990 TI - VEGF-A promotes tissue repair-associated lymphatic vessel formation via VEGFR-2 and the alpha1beta1 and alpha2beta1 integrins. AB - Vascular endothelial growth factor-A (VEGF-A) is strongly up-regulated in wounded cutaneous tissue and promotes repair-associated angiogenesis. However, little is known about its role in lymphatic regeneration of the healing skin. We studied wound healing in transgenic mice that overexpress VEGF-A specifically in the epidermis and in wild-type mice in the absence or presence of inhibitors of VEGF A signaling. Surprisingly, transgenic overexpression of VEGF-A in the skin promoted lymphangiogenesis at the wound healing site, whereas systemic blockade of VEGFR-2 prevented lymphatic vessel formation. Studies in cultured lymphatic endothelial cells revealed that VEGF-A induced expression of the alpha1 and alpha2 integrins, which promoted their in vitro tube formation and their haptotactic migration toward type I collagen. VEGF-A-induced lymphatic endothelial cord formation and haptotactic migration were suppressed by anti alpha1 and anti-alpha2 integrin blocking antibodies, and systemic blockade of the alpha1 and alpha2 integrins inhibited VEGF-A-driven lymphangiogenesis in vivo. We propose that VEGF-A promotes lymphatic vasculature formation via activation of VEGFR-2 and that lineage-specific differences of integrin receptor expression contribute to the distinct dynamics of wound-associated angiogenesis and lymphangiogenesis. PMID- 15132991 TI - Hepatitis B viral HBx induces matrix metalloproteinase-9 gene expression through activation of ERK and PI-3K/AKT pathways: involvement of invasive potential. AB - Hepatitis B virus (HBV) X protein (HBx) has been shown to be essential for the development of hepatocellular carcinoma (HCC). Recently, we have found that HBx causes the progression of liver cancer through down-expression of PTEN, known as a tumor suppressor gene (1). The prognosis for HCC depends mainly on the clinicopathological characteristic regarding invasion and metastasis. The expression of matrix metalloproteinase (MMP)-9 has been implicated as playing an important role in HCC invasion and metastasis. We previously reported that HBV infection increased the invasiveness of hepatocytes and HCC cells through the transcriptional activation of MMP-9 (2). The HBx was shown to activate the mitogen-activated protein (MAP) kinase and phosphatidylinositol 3-kinase (PI-3K) signal cascade, which is essential for activation of transcription factors such as activating protein (AP)-1 and nuclear factor (NF)-kappaB. In this study, we show that the HBx protein stimulates the activities of the PI-3K-Akt/ protein kinase B (PKB) as well as extracellular signal-regulated kinase 1/2 (ERK 1/2) in HBx-transfected cells. Furthermore, we have shown that enhanced expression of MMP 9 in HBx-transfected cells mediated by not only activation of AP-1 transcriptional activity through ERKs pathway but also activation of NF-kappaB transcriptional activity through PI-3K-AKT/PKB pathway, and was associated with the invasive potential. However, treatment with U0126 (known as the ERKs inhibitor) or wortmannin (known as the PI-3K inhibitor), but not SB203580 (known as the p38 MAPK inhibitor), markedly inhibited the expression of MMP-9 induced by HBx in HBx-transfected cells. Seemingly, the invasiveness of HBx-transfected cells was decreased by treating with U0126 or wortmannin, but not SB203580. These results clearly suggest that the HBx contributed to the transcriptional regulation of MMP-9 through the ERKs and PI-3K-AKT/PKB pathway, and increased an invasive potential of cells. PMID- 15132992 TI - Extending the repertoire of the mixed-lineage leukemia gene MLL in leukemogenesis. PMID- 15132993 TI - Pol12, the B subunit of DNA polymerase alpha, functions in both telomere capping and length regulation. AB - The regulation of telomerase action, and its coordination with conventional DNA replication and chromosome end "capping," are still poorly understood. Here we describe a genetic screen in yeast for mutants with relaxed telomere length regulation, and the identification of Pol12, the B subunit of the DNA polymerase alpha (Pol1)-primase complex, as a new factor involved in this process. Unlike many POL1 and POL12 mutations, which also cause telomere elongation, the pol12 216 mutation described here does not lead to either reduced Pol1 function, increased telomeric single-stranded DNA, or a reduction in telomeric gene silencing. Instead, and again unlike mutations affecting POL1, pol12-216 is lethal in combination with a mutation in the telomere end-binding and capping protein Stn1. Significantly, Pol12 and Stn1 interact in both two-hybrid and biochemical assays, and their synthetic-lethal interaction appears to be caused, at least in part, by a loss of telomere capping. These data reveal a novel function for Pol12 and a new connection between DNA polymerase alpha and Stn1. We propose that Pol12, together with Stn1, plays a key role in linking telomerase action with the completion of lagging strand synthesis, and in a regulatory step required for telomere capping. PMID- 15132994 TI - Recruitment of NIMA kinase shows that maturation of the S. pombe spindle-pole body occurs over consecutive cell cycles and reveals a role for NIMA in modulating SIN activity. AB - Mitotic exit in Saccharomyces cerevisiae and septation in Schizosaccharomyces pombe are regulated by a conserved signaling network called the mitotic exit and septum initiation networks (SIN), respectively. The network is active on one of the two anaphase B spindle-pole bodies (SPBs). Whereas the inherent asymmetry of growth by budding accounts for elements of the asymmetry in S. cerevisiae, it has been unclear how, or why, the pathway is asymmetric in S. pombe. We show that elements of SPB duplication in S. pombe are conservative, and that the SIN is active on the new SPB. SIN association with the new SPB persists after transient depolymerization of microtubules. The localization of the NIMA-related kinase, Fin1, reveals further complexity in SPB inheritance. Fin1 associates with the SPB bearing the older components in all cells and with the "new" SPB in half of the population. Fin1 only binds the new SPB when this new SPB has arisen from the duplication of an SPB that is two or more cycles old. Thus, each of the four SPBs generated over two consecutive cell cycles are different, because they have distinct fates in the next cell cycle. Fin1 binds the SPB once the SIN is active and the association requires the SIN inhibitors Byr4 and Cdc16. Fin1 physically associates with Byr4. Compromising Fin1 function leads to SIN activation on both anaphase B SPBs and promotes septation, indicating that Fin1 restrains SIN activity on the old SPB. PMID- 15132995 TI - Positive and negative functions of the SAGA complex mediated through interaction of Spt8 with TBP and the N-terminal domain of TFIIA. AB - A surface that is required for rapid formation of preinitiation complexes (PICs) was identified on the N-terminal domain (NTD) of the RNA Pol II general transcription factor TFIIA. Site-specific photocross-linkers and tethered protein cleavage reagents positioned on the NTD of TFIIA and assembled in PICs identified the SAGA subunit Spt8 and the TFIID subunit Taf4 as located near this surface. In agreement with these findings, mutations in Spt8 and the TFIIA NTD interact genetically. Using purified proteins, it was found that TFIIA and Spt8 do not stably bind to each other, but rather both compete for binding to TBP. Consistent with this competition, Spt8 inhibits the binding of SAGA to PICs in the absence of activator. In the presence of activator, Spt8 enhances transcription in vitro, and the positive function of the TFIIA NTD is largely mediated through Spt8. Our results suggest a mechanism for the previously observed positive and negative effects of Spt8 on transcription observed in vivo. PMID- 15132996 TI - Angiopoietin-1 modulates endothelial cell function and gene expression via the transcription factor FKHR (FOXO1). AB - Despite genetic evidence establishing angiopoietin-1 (Ang-1) as an essential regulator of vascular development, the molecular mechanisms underlying Ang-1 function are almost completely uncharacterized. In this report, we demonstrate that Ang-1, via Akt activation, is a potent inhibitor of the forkhead transcription factor FKHR (FOXO1), identifying for the first time a nuclear signaling pathway through which Ang-1 modulates gene expression. We use microarray analysis to show that FKHR, whose function in endothelial cells has not previously been elucidated, regulates many genes associated with vascular destabilization and remodeling (including angiopoietin-2, an Ang-1 antagonist) and endothelial cell apoptosis (e.g., survivin, TRAIL). Ang-1 inhibits FKHR mediated changes in gene expression and FKHR-induced apoptosis. Analysis of gene expression changes induced by an activated version of Akt confirms that FKHR is a major target through which Akt regulates transcription in endothelial cells. We use RNA interference to demonstrate that FKHR is required for the expression of genes (including Ang-2) that have important vascular functions. Our data suggest a novel, tissue-specific role for the Akt/FKHR pathway in the vasculature and suggest a mechanistic basis for the previously described actions of Ang-1 as a regulator of endothelial cell survival and blood vessel stability. PMID- 15132997 TI - Interactions between Sox9 and beta-catenin control chondrocyte differentiation. AB - Chondrogenesis is a multistep process that is essential for endochondral bone formation. Previous results have indicated a role for beta-catenin and Wnt signaling in this pathway. Here we show the existence of physical and functional interactions between beta-catenin and Sox9, a transcription factor that is required in successive steps of chondrogenesis. In vivo, either overexpression of Sox9 or inactivation of beta-catenin in chondrocytes of mouse embryos produces a similar phenotype of dwarfism with decreased chondrocyte proliferation, delayed hypertrophic chondrocyte differentiation, and endochondral bone formation. Furthermore, either inactivation of Sox9 or stabilization of beta-catenin in chondrocytes also produces a similar phenotype of severe chondrodysplasia. Sox9 markedly inhibits activation of beta-catenin-dependent promoters and stimulates degradation of beta-catenin by the ubiquitination/proteasome pathway. Likewise, Sox9 inhibits beta-catenin-mediated secondary axis induction in Xenopus embryos. Beta-catenin physically interacts through its Armadillo repeats with the C terminal transactivation domain of Sox9. We hypothesize that the inhibitory activity of Sox9 is caused by its ability to compete with Tcf/Lef for binding to beta-catenin, followed by degradation of beta-catenin. Our results strongly suggest that chondrogenesis is controlled by interactions between Sox9 and the Wnt/beta-catenin signaling pathway. PMID- 15132999 TI - Associations between physical activity and bone mass in black and white South African children at age 9 yr. AB - We investigated differences in physical activity (PA) levels between black and white South African 9-yr-old children and their association with bone mineral content (BMC) and density (BMD) by using dual-energy X-ray absorptiometry. PA was analyzed in terms of a metabolic (METPA; weighted metabolic score of intensity, frequency, and duration) and a mechanical (MECHPA; sum of all ground reaction forces multiplied by duration) component. There were significant ethnic differences in patterns of activity. White children expended a significantly greater energy score (METPA of 21.7 +/- 2.9) than black children (METPA of 9.5 +/ 0.5) (P < 0.001). When children were divided into quartiles according to the amount and intensity of sport played, the most active white children (using METPA scores) had significantly higher whole body BMD and higher hip and spine BMC and BMD than less active children. White children in the highest MECHPA quartile also showed significantly higher whole body, hip, and spine BMC and BMD than those children in the lowest quartile. No association between exercise and bone mass of black children was found. In this population, PA has an osteogenic association with white children, but not black children, which may be explained by the lower levels of PA in the black children. Despite this, black children had significantly greater bone mass at the hip and spine (girls only) (P < 0.001) even after adjustment for body size. The role of exercise in increasing bone mass may become increasingly critical as a protective mechanism against osteoporosis in both ethnic groups, especially because the genetic benefit exhibited by black children to higher bone mass may be weakened with time, as environmental influences become stronger. PMID- 15132998 TI - Divergent functions of murine Pax3 and Pax7 in limb muscle development. AB - Pax genes encode evolutionarily conserved transcription factors that play critical roles in development. Pax3 and Pax7 constitute one of the four Pax subfamilies. Despite partially overlapping expression domains, mouse mutations for Pax3 and Pax7 have very different consequences. To investigate the mechanism of these contrasting phenotypes, we replaced Pax3 by Pax7 by using gene targeting in the mouse. Pax7 can substitute for Pax3 function in dorsal neural tube, neural crest cell, and somite development, but not in the formation of muscles involving long-range migration of muscle progenitor cells. In limbs in which Pax3 is replaced by Pax7, the severity of the muscle phenotype increases as the number of Pax7 replacement alleles is reduced, with the forelimb more affected than the hindlimb. We show that this hypomorphic activity of Pax7 is due to defects in delamination, migration, and proliferation of muscle precursor cells with inefficient activation of c-met in the hypaxial domain of the somite. Despite this, overall muscle patterning is retained. We conclude that functions already prefigured by the single Pax3/7 gene present before vertebrate radiation are fulfilled by Pax7 as well as Pax3, whereas the role of Pax3 in appendicular muscle formation has diverged, reflecting the more recent origin of this mode of myogenesis. PMID- 15133001 TI - Passive stretch inhibits central corelike lesion formation in the soleus muscles of hindlimb-suspended unloaded rats. AB - Hindlimb suspension unloading (HSU) is a ground-based model simulating the effects of microgravity unloading on the musculoskeletal system. In this model, gravity causes the hind foot of the rat to drop, opening the front of the ankle to 90-105 degrees plantar flexion at rest. As HSU proceeds, the normal weight bearing angle of 30 degrees dorsiflexion is achieved progressively less, and the contraction range of soleus is abbreviated. Our laboratory reported that 12 days of HSU caused central corelike lesions (CCLs) of myofibril breakdown (Riley DA, Slocum GR, Bain JL, Sedlak FR, Sowa TE, and Mellender JW. J Appl Physiol. 69: 58 66, 1990). The present study investigated whether daily stretch of the calf muscles prevents CCL formation. The soleus muscles of HSU Sprague-Dawley male rats (approximately 287 g) were lengthened by unilateral ankle splinting at 30 degrees. Compared with the nonsplinted side, splinting for 10 or 20 min per day in awake rats significantly decreased CCLs in soleus by 88 and 91%, respectively (P < 0.01). Compared with control muscle wet weight, 20-min splinting reduced atrophy by 33%, whereas 10-min splinting ameliorated atrophy by 17% (P < 0.01). Bilateral soleus electromyograph recording revealed higher levels of contractile activity on the splinted side during splinting. To isolate the effects of stretch from isometric contractile activity, contractions were eliminated by whole animal anesthesia with isoflurane during 10-min daily splinting. The percentage of fibers with CCLs was reduced by 57%, and the average lesion size was 29% smaller in the stretched muscle (P < 0.05). Soleus muscle wet weight and fiber area were unaltered by stretch alone. Loaded contractions during splinting are necessary to prevent muscle fiber atrophy. Passive muscle stretch acts to maintain myofibril structural integrity. PMID- 15133000 TI - Correspondence between laryngeal vocal fold movement and muscle activity during speech and nonspeech gestures. AB - To better understand the role of each of the laryngeal muscles in producing vocal fold movement, activation of these muscles was correlated with laryngeal movement during different tasks such as sniff, cough or throat clear, and speech syllable production. Four muscles [the posterior cricoarytenoid, lateral cricoarytenoid, cricothyroid (CT), and thyroarytenoid (TA)] were recorded with bipolar hooked wire electrodes placed bilaterally in four normal subjects. A nasoendoscope was used to record vocal fold movement while simultaneously recording muscle activity. Muscle activation level was correlated with ipsilateral vocal fold angle for vocal fold opening and closing. Pearson correlation coefficients and their statistical significance were computed for each trial. Significant effects of muscle (P < or = 0.0005) and task (P = 0.034) were found on the r (transformed to Fisher's Z') values. All of the posterior cricoarytenoid recordings related significantly with vocal opening, whereas CT activity was significantly correlated with opening only during sniff. The TA and lateral cricoarytenoid activities were significantly correlated with vocal fold closing during cough. During speech, the CT and TA activity correlated with both opening and closing. Laryngeal muscle patterning to produce vocal fold movement differed across tasks; reciprocal muscle activity only occurred on cough, whereas speech and sniff often involved simultaneous contraction of muscle antagonists. In conclusion, different combinations of muscle activation are used for biomechanical control of vocal fold opening and closing movements during respiratory, airway protection, and speech tasks. PMID- 15133002 TI - Bionic epidural stimulation restores arterial pressure regulation during orthostasis. AB - A bionic baroreflex system (BBS) is a computer-assisted intelligent feedback system to control arterial pressure (AP) for the treatment of baroreflex failure. To apply this system clinically, an appropriate efferent neural (sympathetic vasomotor) interface has to be explored. We examined whether the spinal cord is a candidate site for such interface. In six anesthetized and baroreflex deafferentiated cats, a multielectrode catheter was inserted into the epidural space to deliver epidural spinal cord stimulation (ESCS). Stepwise changes in ESCS rate revealed a linear correlation between ESCS rate and AP for ESCS rates of 2 pulses/s and above (r2, 0.876-0.979; slope, 14.3 +/- 5.8 mmHg.pulses(-1).s; pressure axis intercept, 35.7 +/- 25.9 mmHg). Random changes in ESCS rate with a white noise sequence revealed dynamic transfer function of peripheral effectors. The transfer function resembled a second-order, low-pass filter with a lag time (gain, 16.7 +/- 8.3 mmHg.pulses(-1).s; natural frequency, 0.022 +/- 0.007 Hz; damping coefficient, 2.40 +/- 1.07; lag time, 1.06 +/- 0.41 s). On the basis of the transfer function, we designed an artificial vasomotor center to attenuate hypotension. We evaluated the performance of the BBS against hypotension induced by 60 degrees head-up tilt. In the cats with baroreflex failure, head-up tilt dropped AP by 37 +/- 5 mmHg in 5 s and 59 +/- 11 mmHg in 30 s. BBS with optimized feedback parameters attenuated hypotension to 21 +/- 2 mmHg in 5 s (P < 0.05) and 8 +/- 4 mmHg in 30 s (P < 0.05). These results indicate that ESCS-mediated BBS prevents orthostatic hypotension. Because epidural stimulation is a clinically feasible procedure, this BBS can be applied clinically to combat hypotension associated with various pathophysiologies. PMID- 15133003 TI - Effects of insulin resistance on substrate utilization during exercise in overweight women. AB - During exercise, obese individuals oxidize less glycogen and more fat than their lean counterparts, but the shift in substrate use may be mediated by insulin resistance rather than body fat per se. In addition, individuals with Type 2 diabetes are not resistant to contraction-mediated glucose uptake during exercise, but in vivo studies uncomplicated by hyperglycemia are lacking. The purpose of this study was to compare blood glucose uptake and the balance between carbohydrate and fat utilization during exercise in insulin-resistant (IR) and insulin-sensitive (IS) women of equivalent body fatness and maximal oxygen consumption (VO2 max). Twelve overweight sedentary women were divided into two groups with similar body mass index (IR = 28.5 +/- 1.6, IS = 27.5 +/- 1.9), lean mass (IR = 42.4 +/- 1.8 kg, IS = 41.5 +/- 1.9 kg), and VO2 max (IR = 29.7 +/- 3.5 ml.kg(-1).min(-1), IS = 30.7 +/- 3.9 ml.kg(-1).min(-1)) but a markedly different composite insulin sensitivity index (IR = 3.0 +/- 0.7, IS = 7.7 +/- 0.9). Blood glucose kinetics and substrate oxidation were assessed by stable isotope dilution and indirect calorimetry during 50 min of treadmill walking at 45% VO2 max. Total carbohydrate oxidation and estimated muscle glycogen use were significantly lower in the IR group. Blood glucose uptake was the same in the IR and IS groups. These data suggest that insulin resistance, independent of body fat, spares muscle glycogen and shifts substrate oxidation toward less carbohydrate use during exercise. Insulin-resistant individuals with normoglycemia appear to have no defect in blood glucose uptake during exercise. PMID- 15133004 TI - Low-frequency depression of tension in the cat gastrocnemius muscle after eccentric exercise. AB - Subjecting a muscle to a series of eccentric contractions in which the contracting muscle is lengthened results in a number of changes in its mechanical properties. These include a fall in isometric tension that is particularly pronounced during low-frequency stimulation, a phenomenon known as low-frequency depression (LFD). Reports of LFD have not taken into account the shift in optimum length for active tension generation to longer muscle lengths that takes place after eccentric contractions. Given the length dependence of the stimulation frequency-tension curve, we tested the hypothesis that the change in this relationship after eccentric exercise is due to the shift in optimum length. We measured LFD by recording tension in response to a linearly increasing rate of stimulation of the nerve to medial gastrocnemius of anesthetized cats, over the range 0-100 pulses per second. Tension responses were measured before and after 50 eccentric contractions consisting of 6-mm stretches starting at 3 mm below optimum length and finishing at 3 mm above it. An index of LFD was derived from the tension responses to ramp stimulation. It was found that LFD after the eccentric contractions was partly, but not entirely, due to changes in the muscle's optimum length. An additional factor was the effect of fatigue. These observations led to the conclusion that the muscle length dependence of LFD was reduced by eccentric contractions. All of this means that after eccentric exercise the tension deficit at low rates of muscle activation is likely to be less severe than first thought. PMID- 15133005 TI - Downhill running: a model of exercise hyperemia in the rat spinotrapezius muscle. AB - To utilize the rat spinotrapezius muscle as a model to investigate the microcirculatory consequences of exercise training, it is necessary to design an exercise protocol that recruits this muscle. There is evidence that the spinotrapezius is derecruited during standard treadmill exercise protocols performed on the uphill treadmill (i.e., 6 degrees incline). This investigation tested the hypothesis that downhill running would effectively recruit the spinotrapezius muscle as assessed by the presence of an exercise hyperemia response. We used radioactive 15-microm microspheres to determine blood flows in the spinotrapezius and selected hindlimb muscles of female Sprague-Dawley rats at rest and during downhill (i.e., -14 degrees incline; 331 +/- 5 g body wt, n = 7) and level (i.e., 0 degrees incline; 320 +/- 11 g body wt, n = 5) running at 30 m/min. Both level and downhill exercise increased blood flow to all hindlimb muscles (P < 0.01). However, in marked contrast to the absence of a hyperemic response to level running, blood flow to the spinotrapezius muscle increased from 26 +/- 6 ml.min(-1).100 g(-1) at rest to 69 +/- 8 ml.min(-1).100 g(-1) during downhill running (P < 0.01). These findings indicate that downhill running represents an exercise paradigm that recruits the spinotrapezius muscle and thereby constitutes a tenable physiological model for investigating the adaptations induced by exercise training (i.e., the mechanisms of altered microcirculatory control by transmission light microscopy). PMID- 15133006 TI - Decreased exhaled nitric oxide as a marker of postinsult immune paralysis. AB - Nitric oxide (NO) regulates neutrophil migration and alveolar macrophage functions such as cytokine synthesis and bacterial killing, both of which are impaired in immune paralysis associated with critical illness. The aim of this study was to determine whether NO is involved in immune paralysis and whether exhaled NO measurement could help to monitor pulmonary defenses. NO production (protein expression, enzyme activity, end products, and exhaled NO measurements) was assessed in rats after cecal ligation and puncture to induce a mild peritonitis (leading to approximately 20% mortality rate). An early and sustained decrease in exhaled NO was found after peritonitis (from 1 to 72 h) compared with healthy rats [median (25th-75th percentile), 1.5 parts per billion (ppb) (1.2 1.7) vs. 4.0 ppb (3.6-4.3), P < 0.05], despite increased NO synthase-2 and unchanged NO synthase-3 protein expression in lung tissue. NO synthase-2 activity was decreased in lung tissue. Nitrites and nitrates in supernatants of isolated alveolar macrophages decreased after peritonitis compared with healthy rats, and an inhibitory experiment suggested arginase overactivity in alveolar macrophages bypassing the NO substrate. Administration of the NO synthase-2 inhibitor aminoguanidine to healthy animals reproduced the decreased neutrophil migration toward alveolar spaces that was observed after peritonitis, but L-arginine administration after peritonitis failed to correct the defect of neutrophil emigration despite increasing exhaled NO compared with D-arginine administration [4.8 (3.9-5.7) vs. 1.6 (1.3-1.7) ppb, respectively, P < 0.05]. In conclusion, the decrease in exhaled NO observed after mild peritonitis could serve as a marker for lung immunodepression. PMID- 15133007 TI - Facilitation of the diaphragm response to transcranial magnetic stimulation by increases in human respiratory drive. AB - The human respiratory neural drive has an automatic component (bulbospinal pathway) and a volitional component (corticospinal pathway). The aim of this study was to assess the effects of a hypercapnia-induced increase in the automatic respiratory drive on the function of the diaphragmatic corticospinal pathway as independently as possible of any other influence. Thirteen healthy volunteers breathed room air and then 5 and 7% hyperoxic CO2. Cervical (cms) and transcranial (tms) magnetic stimulations were performed during early inspiration and expiration. Transdiaphragmatic pressure (Pdi) and surface electromyogram of the diaphragm (DiEMG) and of the abductor pollicis brevis (apbEMG) were recorded in response to cms and tms. During inspiration, Pdi,cms was unaffected by CO2, but Pdi,tms increased significantly with 7% CO2. During expiration, Pdi,cms was significantly reduced by CO2, whereas Pdi,tms was preserved. DiEMG,tms latencies decreased significantly during early inspiration and expiration (air vs. 5% CO2 and air vs. 7% CO2). DiEMG,tms amplitude increased significantly in response to early expiration-tms (air vs. 5% CO2 and air vs. 7% CO2) but not in response to early inspiration-tms. DiEMG,cms latencies and amplitudes were not affected by CO2 whereas 7% CO2 significantly increased the apbEMG,cms latency. The apbEMG,tms vs. apbEMG,cms latency difference was unaffected by CO2. In conclusion, increasing the automatic drive to breathe facilitates the response of the diaphragm to tms, during both inspiration and expiration. This could allow the corticospinal drive to breathe to keep the capacity to modulate respiration in conditions under which the automatic respiratory control is stimulated. PMID- 15133008 TI - Expression of MHC-beta and MCT1 in cardiac muscle after exercise training in myocardial-infarcted rats. AB - To evaluate the hypothesis that increasing the potential for glycolytic metabolism would benefit the functioning of infarcted myocardium, we investigated whether mild exercise training would increase the activities of oxidative enzymes, expression of carbohydrate-related transport proteins (monocarboxylate transporter MCT1 and glucose transporter GLUT4), and myosin heavy chain (MHC) isoforms. Myocardial infarction (MI) was produced by occluding the proximal left coronary artery in rat hearts for 30 min. After the rats performed 6 wk of run training on a treadmill, the wall of the left ventricle was dissected and divided into the anterior wall (AW; infarcted region) and posterior wall (PW; noninfarcted region). MI impaired citrate synthase and 3-hydroxyacyl-CoA dehydrogenase activities in the AW (P < 0.01) but not in the noninfarcted PW. No differences in the expression of MCT1 were found in either tissues of AW and PW after MI, whereas exercise training significantly increased the MCT1 expression in all conditions, except AW in the MI rats. Exercise training resulted in an increased expression of GLUT4 protein in the AW in the sham rats and in the PW in the MI rats. The relative amount of MHC-beta was significantly increased in the AW and PW in MI rats compared with sham rats. However, exercise training resulted in a significant increase of MHC-alpha expression in both AW and PW in both sham and MI rats (P < 0.01). These findings suggest that mild exercise training enhanced the potential for glycolytic metabolism and ATPase activity of the myocardium, even in the MI rats, ensuring a beneficial role in the remodeling of the heart. PMID- 15133009 TI - Effects of prior heavy-intensity exercise on pulmonary O2 uptake and muscle deoxygenation kinetics in young and older adult humans. AB - Pulmonary O2 uptake (VO2p) and muscle deoxygenation kinetics were examined during moderate-intensity cycling (80% lactate threshold) without warm-up and after heavy-intensity warm-up exercise in young (n = 6; 25 +/- 3 yr) and older (n = 5; 68 +/- 3 yr) adults. We hypothesized that heavy warm-up would speed VO2p kinetics in older adults consequent to an improved intramuscular oxygenation. Subjects performed step transitions (n = 4; 6 min) from 20 W to moderate-intensity exercise preceded by either no warm-up or heavy-intensity warm-up (6 min). VO2p was measured breath by breath. Oxy-, deoxy-(HHb), and total hemoglobin and myoglobin (Hb(tot)) of the vastus lateralis muscle were measured continuously by near-infrared spectroscopy (NIRS). VO2p (phase 2; tau) and HHb data were fit with a monoexponential model. After heavy-intensity warm-up, oxyhemoglobin (older subjects: 13 +/- 9 microM; young subjects: 9 +/- 8 microM) and Hb(tot) (older subjects: 12 +/- 8 microM; young subjects: 14 +/- 10 microM) were elevated (P < 0.05) relative to the no warm-up pretransition baseline. In older adults, tauVO2p adapted at a faster rate (P < 0.05) after heavy warm-up (30 +/- 7 s) than no warm up (38 +/- 5 s), whereas in young subjects, tauVO2p was similar in no warm-up (26 +/- 7 s) and heavy warm-up (25 +/- 5 s). HHb adapted at a similar rate in older and young adults after no warm-up; however, in older adults after heavy warm-up, the adaptation of HHb was slower (P < 0.01) compared with young and no warm-up. These data suggest that, in older adults, VO2p kinetics may be limited by a slow adaptation of muscle blood flow and O2 delivery. PMID- 15133010 TI - Skeletal muscle oxidative metabolism in sedentary humans: 31P-MRS assessment of O2 supply and demand limitations. AB - Previously, it was demonstrated in exercise-trained humans that phosphocreatine (PCr) recovery is significantly altered by fraction of inspired O2 (FI(O2)), suggesting that in this population under normoxic conditions, O2 availability limits maximal oxidative rate. Haseler LJ, Hogan ML, and Richardson RS. J Appl Physiol 86: 2013-2018, 1999. To further elucidate these population-specific limitations to metabolic rate, we used 31P-magnetic resonance spectroscopy to study the exercising human gastrocnemius muscle under conditions of varied FI(O2) in sedentary subjects. To test the hypothesis that PCr recovery from submaximal exercise in sedentary subjects is not limited by O2 availability, but rather by their mitochondrial capacity, six sedentary subjects performed three bouts of 6 min steady-state submaximal plantar flexion exercise followed by 5 min of recovery while breathing three different FI(O2) (0.10, 0.21, and 1.00). PCr recovery time constants were significantly longer in hypoxia (47.0 +/- 3.2 s), but there was no difference between hyperoxia (31.8 +/- 1.9 s) and normoxia (30.0 +/- 2.1 s) (mean +/- SE). End-exercise pH was not significantly different across treatments. These results suggest that the maximal muscle oxidative rate of these sedentary subjects, unlike their exercise-trained counterparts, is limited by mitochondrial capacity and not O2 availability in normoxia. Additionally, the significant elongation of PCr recovery in these subjects in hypoxia illustrates the reliance on O2 supply at the other end of the O2 availability spectrum in both sedentary and active populations. PMID- 15133011 TI - Heterogeneity of airway hyperresponsiveness: time for unconventional, but traditional, studies. PMID- 15133012 TI - Ventrolateral medullary respiratory network participation in the expiration reflex in the cat. AB - The expiration reflex is a distinct airway defensive response characterized by a brief, intense expiratory effort and coordinated adduction and abduction of the laryngeal folds. This study addressed the hypothesis that the ventrolateral medullary respiratory network participates in the reflex. Extracellular neuron activity was recorded with microelectrode arrays in decerebrated, neuromuscular blocked, ventilated cats. In 32 recordings (17 cats), 232 neurons were monitored in the rostral (including Botzinger and pre-Botzinger complexes) and caudal ventral respiratory group. Neurons were classified by firing pattern, evaluated for spinal projections, functional associations with recurrent laryngeal and lumbar nerves, and firing rate changes during brief, large increases in lumbar motor nerve discharge (fictive expiration reflex, FER) elicited during mechanical stimulation of the vocal folds. Two hundred eight neurons were respiratory modulated, and 24 were nonrespiratory; 104 of the respiratory and 6 of the nonrespiratory-modulated neurons had altered peak firing rates during the FER. Increased firing rates of bulbospinal neurons and expiratory laryngeal premotor and motoneurons during the expiratory burst of FER were accompanied by changes in the firing patterns of putative propriobulbar neurons proposed to participate in the eupneic respiratory network. The results support the hypothesis that elements of the rostral and caudal ventral respiratory groups participate in generating and shaping the motor output of the FER. A model is proposed for the participation of the respiratory network in the expiration reflex. PMID- 15133013 TI - Adrenalectomy in mice does not prevent loss of intestinal lymphocytes after exercise. AB - Exhaustive exercise is associated with an increase in circulating glucocorticoids (GCs), lymphocyte apoptosis, and a reduction in intestinal lymphocyte number. The present study examined the role of GCs on the numerical changes seen in intestinal lymphocytes after exercise. Female C57BL/6 mice were bilaterally adrenalectomized (ADX; n = 18) or given sham surgery (Sham; n = 18) and assigned to one of three exercise conditions: treadmill running (28 m/min, 90 min, 2 degrees slope) and killed immediately or after 24 h recovery, or not exercised and killed immediately after 90-min exposure to the treadmill environment. Lymphocytes were isolated from the intestines with CD45(+) cells collected by positive selection using magnetic bead separation columns, and lymphocyte subpopulations were analyzed by flow cytometry for CD45(+), CD3alphabeta(+), CD3gammadelta(+), CD8beta(+), CD8alpha(+), CD4(+), and NK(+) phenotypic markers. ADX mice had significantly more intestinal CD45(+) leukocytes (P < 0.05) and CD3alphabeta(+) (P < 0.05), CD3gammadelta(+) (P < 0.01), CD8alpha(+) (P < 0.001), and NK(+) (P < 0.05) intestinal lymphocytes than Sham mice. There was a significant effect of exercise condition on total intestinal CD45(+) leukocytes (P < 0.01) and CD3alphabeta(+) (P < 0.05), CD8alpha(+) (P < 0.001), and CD4(+) (P < 0.05) intestinal lymphocytes, with fewer cells at 24 h postexercise compared with the other treatment conditions. There were no surgical x exercise interaction effects on the CD3 and CD8 phenotype numbers. Plasma corticosterone was virtually nil in ADX mice regardless of exercise condition but was significantly elevated in Sham mice immediately postexercise (P < 0.001). The data indicate that ADX does not prevent the loss of lymphocytes from the intestinal mucosa 24 h after strenuous exercise and GCs are not directly causal in the leukopenia of exercise. PMID- 15133014 TI - Gas exchange during separate diaphragm and intercostal muscle breathing. AB - In patients with diaphragm paralysis, ventilation to the basal lung zones is reduced, whereas in patients with paralysis of the rib cage muscles, ventilation to the upper lung zones in reduced. Inspiration produced by either rib cage muscle or diaphragm contraction alone, therefore, may result in mismatching of ventilation and perfusion and in gas-exchange impairment. To test this hypothesis, we assessed gas exchange in 11 anesthetized dogs during ventilation produced by either diaphragm or intercostal muscle contraction alone. Diaphragm activation was achieved by phrenic nerve stimulation. Intercostal muscle activation was accomplished by electrical stimulation by using electrodes positioned epidurally at the T(2) spinal cord level. Stimulation parameters were adjusted to provide a constant tidal volume and inspiratory flow rate. During diaphragm (D) and intercostal muscle breathing (IC), mean arterial Po(2) was 97.1 +/- 2.1 and 88.1 +/- 2.7 Torr, respectively (P < 0.01). Arterial Pco(2) was lower during D than during IC (32.6 +/- 1.4 and 36.6 +/- 1.8 Torr, respectively; P < 0.05). During IC, oxygen consumption was also higher than that during D (0.13 +/- 0.01 and 0.09 +/- 0.01 l/min, respectively; P < 0.05). The alveolar-arterial oxygen difference was 11.3 +/- 1.9 and 7.7 +/- 1.0 Torr (P < 0.01) during IC and D, respectively. These results indicate that diaphragm breathing is significantly more efficient than intercostal muscle breathing. However, despite marked differences in the pattern of inspiratory muscle contraction, the distribution of ventilation remains well matched to pulmonary perfusion resulting in preservation of normal gas exchange. PMID- 15133015 TI - Fractal branching pattern of the monopodial canine airway. AB - Unlike the human lung, monopodial canine airway branching follows an irregular dichotomized pattern with fractal features. We studied three canine airway molds and found a self-similarity feature from macro- to microscopic scales, which formed a fractal set up to seven scales in the airways. At each fractal scale, lateral branches evenly lined up along an approximately straight main trunk to form three to four two-dimensional structures, and each lateral branch was the monopodial main trunk of the next fractal scale. We defined this pattern as the fractal main lateral-branching pattern, which exhibited similar structures from macro- to microscopic scales, including lobes, sublobes, sub-sublobes, etc. We speculate that it, rather than a mother-daughter pattern, could better describe the actual asymmetrical architecture of the monopodial canine airway. PMID- 15133016 TI - Hand function: peripheral and central constraints on performance. AB - The hand is one of the most fascinating and sophisticated biological motor systems. The complex biomechanical and neural architecture of the hand poses challenging questions for understanding the control strategies that underlie the coordination of finger movements and forces required for a wide variety of behavioral tasks, ranging from multidigit grasping to the individuated movements of single digits. Hence, a number of experimental approaches, from studies of finger movement kinematics to the recording of electromyographic and cortical activities, have been used to extend our knowledge of neural control of the hand. Experimental evidence indicates that the simultaneous motion and force of the fingers are characterized by coordination patterns that reduce the number of independent degrees of freedom to be controlled. Peripheral and central constraints in the neuromuscular apparatus have been identified that may in part underlie these coordination patterns, simplifying the control of multi-digit grasping while placing certain limitations on individuation of finger movements. We review this evidence, with a particular emphasis on how these constraints extend through the neuromuscular system from the behavioral aspects of finger movements and forces to the control of the hand from the motor cortex. PMID- 15133017 TI - Probing the human vestibular system with galvanic stimulation. AB - Galvanic vestibular stimulation (GVS) is a simple, safe, and specific way to elicit vestibular reflexes. Yet, despite a long history, it has only recently found popularity as a research tool and is rarely used clinically. The obstacle to advancing and exploiting GVS is that we cannot interpret the evoked responses with certainty because we do not understand how the stimulus acts as an input to the system. This paper examines the electrophysiology and anatomy of the vestibular organs and the effects of GVS on human balance control and develops a model that explains the observed balance responses. These responses are large and highly organized over all body segments and adapt to postural and balance requirements. To achieve this, neurons in the vestibular nuclei receive convergent signals from all vestibular receptors and somatosensory and cortical inputs. GVS sway responses are affected by other sources of information about balance but can appear as the sum of otolithic and semicircular canal responses. Electrophysiological studies showing similar activation of primary afferents from the otolith organs and canals and their convergence in the vestibular nuclei support this. On the basis of the morphology of the cristae and the alignment of the semicircular canals in the skull, rotational vectors calculated for every mode of GVS agree with the observed sway. However, vector summation of signals from all utricular afferents does not explain the observed sway. Thus we propose the hypothesis that the otolithic component of the balance response originates from only the pars medialis of the utricular macula. PMID- 15133019 TI - Locomotion-respiration coupling: an account of the underlying dynamics. PMID- 15133020 TI - Update on mechanism and catalytic regulation in the NO synthases. PMID- 15133021 TI - Common mechanism of ligand recognition by group II/III WW domains: redefining their functional classification. AB - WW domain is a well known protein module that mediates protein to protein interactions by binding to proline-containing ligands. Based on the ligand predilections, the WW domains have been classified into four major groups. Group II and III WW domains have been reported to bind the proline-leucine and proline arginine motifs, respectively. In the present study, using surface plasmon resonance technique we have shown that these WW domains have almost indistinguishable ligand preferences and kinetic properties. Hence, we propose that Group II and III WW domains should be joined together as one group (Group II/III). Unlike Group I and IV WW domains, Group II/III WW domains can bind simple polyprolines as well as the proline-leucine and proline-arginine motifs, and they possess two Xaa-proline (where Xaa is any amino acid) binding grooves similar to SH3 domains. Our work assigns Group II and III WW domains to a larger family of polyproline-binding modules and proteins, which includes SH3 domains and profilin. Because polyprolines belong to the most frequently found peptide motifs in several genomes, our study implies the versatile importance of Group II/III WW domains in signaling. PMID- 15133022 TI - 3-hydroxypyridine chromophores are endogenous sensitizers of photooxidative stress in human skin cells. AB - Photocarcinogenesis and photoaging are established consequences of chronic exposure of human skin to solar irradiation. Accumulating evidence supports a causative involvement of UVA irradiation in skin photo-damage. UVA photodamage has been attributed to photosensitization by endogenous skin chromophores leading to the formation of reactive oxygen species and organic free radicals as key mediators of cellular photooxidative stress. In this study, 3-hydroxypyridine derivatives contained in human skin have been identified as a novel class of potential endogenous photosensitizers. A structure-activity relationship study of skin cell photosensitization by endogenous pyridinium derivatives (pyridinoline, desmosine, pyridoxine, pyridoxamine, pyridoxal, pyridoxal-5'-phosphate) and various synthetic hydroxypyridine isomers identified 3-hydroxypyridine and N alkyl-3-hydroxypyridinium cation as minimum phototoxic chromophores sufficient to effect skin cell sensitization toward UVB and UVA, respectively. Photosensitization of cultured human skin keratinocytes (HaCaT) and fibroblasts (CF3) by endogenous and synthetic 3-hydroxypyridine derivatives led to a dose dependent inhibition of proliferation, cell cycle arrest in G2/M, and induction of apoptosis, all of which were reversible by thiol antioxidant intervention. Enhancement of UVA-induced intracellular peroxide formation and p38 mitogen activated protein kinase-dependent stress signaling suggest a photooxidative mechanism of skin cell photosensitization by 3-hydroxypyridine derivatives. 3 hydroxypyridine derivatives were potent photosensitizers of macromolecular damage, effecting protein (RNase A) photocross-linking and peptide (melittin) photooxidation with incorporation of molecular oxygen. Based on these results, we conclude that 3-hydroxypyridine derivatives comprising a wide range of skin biomolecules, such as enzymatic collagen cross-links, B6 vitamers, and probably advanced glycation end products in chronologically aged skin constitute a novel class of UVA photosensitizers, capable of skin photooxidative damage. PMID- 15133023 TI - Close functional coupling between Ca2+ release-activated Ca2+ channels, arachidonic acid release, and leukotriene C4 secretion. AB - In non-excitable cells, one major route for Ca2+ influx is through store-operated Ca2+ channels in the plasma membrane. These channels are activated by the emptying of intracellular Ca2+ stores, and in some cell types, particularly of hemopoietic origin, store-operated influx occurs through Ca2+ release-activated Ca2+ (CRAC) channels. However, little is known about the downstream consequences of CRAC channel activation. Here, we report that Ca2+ entry through CRAC channels stimulates arachidonic acid production, whereas Ca2+ release from the stores is ineffective even though the latter evokes a robust intracellular Ca2+ signal. We find that arachidonic acid released by Ca2+ entering through CRAC channels is used to synthesize the potent paracrine proinflammatory signal leukotriene C4 (LTC4). Both pharmacological inhibitors of CRAC channels and mitochondrial depolarization, which impairs CRAC channel activity, suppress arachidonic acid release and LTC4 secretion. Thus, arachidonic acid release is preferentially stimulated by elevated subplasmalemmal Ca2+ levels due to open CRAC channels, suggesting that the enzyme is located close to the CRAC channels. Our results also identify a novel role for CRAC channels, namely the activation of a downstream signal transduction pathway resulting in the secretion of LTC4. Finally, mitochondria are key determinants of the generation of both intracellular (arachidonic acid) and paracrine (LTC4) signals through their effects on CRAC channel activity. PMID- 15133024 TI - Evidence for a role of MSK1 in transforming growth factor-beta-mediated responses through p38alpha and Smad signaling pathways. AB - Smad proteins are central mediators of the transforming growth factor-beta (TGF beta) superfamily signaling. The mitogen-activated protein kinase (MAPK) p38 is also one of the downstream targets required for TGF-beta-mediated responses. Although the interplay between the p38 and Smad signaling pathways might allow cells to display diverse patterns of responses to TGF-beta, the mechanism of this cross-talk is not well established. We report here that inhibition of the p38alpha isoform suppressed the ability of Smad3 to mediate TGF-beta-induced transcriptional responses. The inhibition of p38 activity blocked TGF-beta mediated phosphorylation of the MSK1 kinase, a substrate of p38 that plays an important role in the remodeling of chromatin. Moreover, we observed that expression of dominant-interfering mutants of MSK1 blocked the binding of Smad3 to the coactivator p300 in response to TGF-beta signaling. These data reveal a new mechanism whereby the Smad signaling pathway and the p38 cascade are integrated in the nucleus to activate gene expression. PMID- 15133025 TI - Roles of conserved P domain residues and Mg2+ in ATP binding in the ground and Ca2+-activated states of sarcoplasmic reticulum Ca2+-ATPase. AB - Residues in conserved motifs (625)TGD, (676)FARXXPXXK, and (701)TGDGVND in domain P of sarcoplasmic reticulum Ca(2+)-ATPase, as well as in motifs (601)DPPR and (359)NQR(/K)MSV in the hinge segments connecting domains N and P, were examined by mutagenesis to assess their roles in nucleotide and Mg(2+) binding and stabilization of the Ca(2+)-activated transition state for phosphoryl transfer. In the absence of Mg(2+), mutations removing the charges of domain P residues Asp(627), Lys(684), Asp(703), and Asp(707) increased the affinity for ATP and 2',3'-O-(2,4,6-trinitrophenyl)-8-azidoadenosine 5'-triphosphate. These mutations, as well as Gly(626)--> Ala, were inhibitory for ATP binding in the presence of Mg(2+) and for tight binding of the beta,gamma-bidentate chromium(III) complex of ATP. The hinge mutations had pronounced, but variable, effects on ATP binding only in the presence of Mg(2+). The data demonstrate an unfavorable electrostatic environment for binding of negatively charged nucleotide in domain P and show that Mg(2+) is required to anchor the phosphoryl group of ATP at the phosphorylation site. Mutants Gly(626) --> Ala, Lys(684) --> Met, Asp(703) --> Ala/Ser/Cys, and mutants with alteration to Asp(707) exhibited very slow or negligible phosphorylation, making it possible to measure ATP binding in the pseudo-transition state attained in the presence of both Mg(2+) and Ca(2+). Under these conditions, ATP binding was almost completely blocked in Gly(626) --> Ala and occurred with 12- and 7-fold reduced affinities in Asp(703) --> Ala and Asp(707) --> Cys, respectively, relative to the situation in the presence of Mg(2+) without Ca(2+), whereas in Lys(684) --> Met and Asp(707) --> Ser/Asn the affinity was enhanced 14- and 3-5-fold, respectively. Hence, Gly(626) and Asp(703) seem particularly critical for mediating entry into the transition state for phosphoryl transfer upon Ca(2+) binding at the transport sites. PMID- 15133026 TI - Casein kinase Iepsilon plays a functional role in the transforming growth factor beta signaling pathway. AB - The transforming growth factor-beta (TGF-beta) signaling pathway is known to be involved in a wide range of biological events, including development, cellular differentiation, apoptosis, and oncogenesis. The TGF-beta signal is mediated by ligand binding to the type II receptor, leading to the recruitment and activation of the type I receptor, and subsequent activation of a family of intracellular signal transducing proteins called Smads. Here we report a regulatory role for casein kinase Iepsilon (CKIepsilon) in the TGF-beta signaling cascade. We find that CKIepsilon binds to all Smads and the cytoplasmic domains of the type I and type II receptors both in vitro and in vivo. The interaction of CKIepsilon with the type I and type II receptors is independent of TGF-beta stimulation, whereas the CKIepsilon/Smad interaction is transiently disrupted by ligand treatment. Additionally, CKIepsilon is able to phosphorylate the receptor-activated Smads (Smads 1-3 and 5) and the type II receptor in vitro. Transcriptional reporter assays reveal that transient overexpression of wild type CKIepsilon dramatically reduces basal reporter activity but enhances TGF-beta-stimulated transcription. Furthermore, overexpression of a kinase-dead mutant of CKIepsilon inhibits both basal and ligand-induced transcription, whereas inhibition of endogenous CKI catalytic activity with IC261 blocks only TGF-beta-stimulated reporter activity. Finally, knocking down CKIepsilon protein levels results in a significant increase in basal and TGF-beta-induced transcription. These results suggest that CKIepsilon plays a ligand-dependent, differential, and dual regulatory role within the TGF-beta signaling pathway. PMID- 15133027 TI - Differentiation-dependent sensitivity to apoptogenic factors in PC12 cells. AB - We have investigated the role of the mitochondrial pathway during cell death following serum and nerve growth factor (NGF)/dibutyryl cyclic AMP (Bt(2)cAMP) withdrawal in undifferentiated or NGF/Bt(2)cAMP-differentiated PC12 cells, respectively. Holocytochrome c, Smac/DIABLO, and Omi/HtrA2 are released rapidly following trophic factor deprivation in PC12 cells. Bcl-2 and Akt inhibited this release. The protection, however, persisted longer in differentiated PC12 cells. In differentiated, but not undifferentiated cells, Bcl-2 and Akt also inhibited apoptosis downstream of holocytochrome c release. Thus, undifferentiated PC12 cells showed marked sensitivity to induction of apoptosis by microinjected cytochrome c even in the presence of NGF, Bcl-2, or Akt. In contrast, in differentiated cells these factors suppressed cell death. Consistent with these observations, in vitro processing of procaspase 9 in response to cytochrome c was observed in extracts from undifferentiated but not differentiated cells expressing Akt or Bcl-2. Endogenous caspase 9 was cleaved during cell death, whereas dominant negative caspase 9 inhibited cell death. The results from determining the role of inhibitors of apoptosis (IAPs) suggest that acquisition of inhibition by IAPs is part of the differentiation program. Ubiquitin-DeltaN AVPI Smac/DIABLO induced cell death in differentiated cells only. c-IAP-2 is unregulated in differentiated cells, whereas X-linked IAP levels decreased in these cells coincident with cell death. Moreover, expressing X-linked IAP rendered undifferentiated cells resistant to microinjected cytochrome c. Overall, the inhibitory regulation, of cell death at the level of release of mitochondrial apoptogenic factors and at post-mitochondrial activation of caspase 9 observed in differentiated PC12 cells, is reduced or absent in the undifferentiated counterparts. PMID- 15133028 TI - Ligand selectivity and affinity of chemokine receptor CXCR1. Role of N-terminal domain. AB - Glu-Leu-Arg ("ELR") CXC chemokines interleukin-8 (IL-8) and melanoma growth stimulatory activity (MGSA) recruit neutrophils by binding and activating two receptors, CXCR1 and CXCR2. CXCR1 is specific, binding only IL-8 with nanomolar affinity, whereas CXCR2 is promiscuous, binding all ELRCXC chemokines with high affinity. Receptor signaling consists of two events: interactions between the ligand N-terminal loop (N-loop) and receptor N-terminal domain (N-domain) residues (site I), and between the ligand N-terminal ELR and the receptor juxtamembrane domain (J-domain) residues (site II). It is not known how these interactions mediate ligand affinity and selectivity, and whether binding at one site influences binding and function at the other. Sequence analysis and structure-function studies have suggested that the receptor N-domain plays an important role in ligand selectivity. Here, we report ligand-binding properties and structural characteristics of the CXCR1 N-domain in solution and in detergent micelles that mimic the native membrane environment. We find that IL-8 binds the N-domain with significantly higher affinity in micelles than in solution (approximately 1 microM versus approximately 20 microM) and that MGSA does not bind the N-domain in solution but does in micelles with appreciable affinity (approximately 3 microM). We find that the N-domain is structured in micelles and that the entire N-domain interacts with the micelle in an extended fashion. We conclude that the micellar environment constrains the N-domain, and this conformational restraint influences its ligand-binding properties. Most importantly, our data suggest that for both ligands, site I interaction provides similar affinity and that differential coupling between site I and II interactions is responsible for the observed differences in affinity. PMID- 15133029 TI - FXYD7, mapping of functional sites involved in endoplasmic reticulum export, association with and regulation of Na,K-ATPase. AB - The brain-specific FXYD7 is a member of the recently defined FXYD family that associates with the alpha1-beta1 Na,K-ATPase isozyme and induces an about 2-fold decrease in its apparent K+ affinity. By using the Xenopus oocyte as an expression system, we have investigated the role of conserved and FXYD7-specific amino acids in the cellular routing of FXYD7 and in its association with and regulation of Na,K-ATPase. In contrast to FXYD2 and FXYD4, the studies on FXYD7 show that the conserved FXYD motif in the extracytoplasmic domain is not involved in the efficient association of FXYD7 with Na,K-ATPase. On the other hand, the conserved Gly40 and Gly29, located on the same face of the transmembrane helix, were found to be implicated both in the association with and the regulation of Na,K-ATPase. Mutational analysis of FXYD7-specific regions revealed the presence of an ER export signal at the end of the cytoplasmic tail. Deletion of a C terminal valine residue in FXYD7 significantly delayed and decreased its O glycosylation processing and retarded the rate of its cell surface expression. This result indicates that the C-terminal valine residue is involved in the rapid and selective ER export of FXYD7, which could explain the observed post translational association of FXYD7 with Na,K-ATPase. In conclusion, our study on FXYD7 provides new information on structural determinants of general importance for FXYD protein action. Moreover, FXYD7 is identified as a new member of proteins with a regulated ER export, which suggests that, among FXYD proteins, FXYD7 has a particular regulatory function in brain. PMID- 15133030 TI - Modified heparin inhibits P-selectin-mediated cell adhesion of human colon carcinoma cells to immobilized platelets under dynamic flow conditions. AB - Accumulating evidence indicates that the formation of tumor cell platelet emboli complexes in the blood stream is a very important step during metastases and that the anti-metastasis effects of heparin are partially due to a blockade of P selectin on platelets. In this study, heparin and chemically modified heparins were tested as inhibitors of three human colon carcinoma cell lines (COLO320, LS174T, and CW-2) binding to P-selectin, adhering to CHO cells expressing a transfected human P-selectin cDNA, and adhering to surface-anchored platelets expressing P-selectin under static and flow conditions. The aim was to screen for heparin derivatives with high anti-adhesion activity but negligible anticoagulant activity. In this study, four modified heparins with high anti-adhesion activity were identified including RO-heparin, CR-heparin, 2/3ODS-heparin, and N/2/3DS heparin. NMR analysis proved the reliability of structure of the four modified heparins. Our findings suggested that the 6-O-sulfate group of glucosamine units in heparin is critical for the inhibition of P-selectin-mediated tumor cell adhesion. Heparan sulfate-like proteoglycans on these tumor cell surfaces are implicated in adhesion of the tumor cells to P-selectin. Some chemically modified heparins with low anticoagulant activities, such as 2/3ODS-heparin, may have potential value as therapeutic agents that block P-selectin-mediated cell adhesion and prevent tumor metastasis. PMID- 15133031 TI - A novel role for the immunophilin FKBP52 in copper transport. AB - FK506-binding protein 52 (FKBP52) is an immunophilin that possesses peptidylprolyl cis/trans-isomerase (PPIase) activity and is a component of a subclass of steroid hormone receptor complexes. Several recent studies indicate that immunophilins can regulate neuronal survival and nerve regeneration although the molecular mechanisms are poorly understood. To investigate the function of FKBP52 in the nervous system, we employed a yeast two-hybrid strategy using the PPIase domain (domain I) as bait to screen a neonatal rat dorsal root ganglia cDNA expression library. We identified an interaction between FKBP52 domain I and Atox1, a copper-binding metallochaperone. Atox1 interacts with Menkes disease protein and Wilson disease protein (WD) and functions in copper efflux. The interaction between FKBP52 and Atox1 was observed in both glutathione S transferase pull-down experiments and when proteins were ectopically expressed in human embryonic kidney (HEK) 293T cells and was sensitive to FK506. Interestingly, the FKBP52/Atox1 interaction was enhanced when HEK 293T cells were cultured in copper-supplemented medium and decreased in the presence of the copper chelator, bathocuproine disulfate, suggesting that the interaction is regulated in part by intracellular copper. Overexpression of FKBP52 increased rapid copper efflux in (64)Cu-loaded cells, as did the overexpression of WD transporter. Taken together, our present findings suggest that FKBP52 is a component of the copper efflux machinery, and in so, may also promote neuroprotection from copper toxicity. PMID- 15133032 TI - Interleukin-7 and transforming growth factor-beta play counter-regulatory roles in protein kinase C-delta-dependent control of fibroblast collagen synthesis in pulmonary fibrosis. AB - Transforming growth factor-beta (TGF-beta) is a potent fibrogenic factor responsible for promoting synthesis of extracellular matrix. Interleukin-7 (IL-7) inhibits TGF-beta signaling by up-regulating Smad7, a major inhibitor of the Smad family. In a variety of cells, TGF-beta-mediated activation of target genes requires active protein kinase C-delta (PKC-delta) in addition to Smads (1). We determined the role of PKC-delta in the regulation of pulmonary fibroblast collagen synthesis in response to TGF-beta and IL-7 stimulation. Here we show that TGF-beta and IL-7 have opposing effects on PKC-delta; TGF-beta stimulates, while IL-7 inhibits, PKC-delta activity. IL-7 inhibits TGF-beta-induced PKC-delta phosphorylation at Ser-645 and Thr-505. Inhibition of PKC-delta with specific small inhibitory RNA restores TGF-beta-mediated induction of Smad7 and in parallel significantly reduces TGF-beta-mediated collagen synthesis. Thus, PKC delta may play a critical role in the pathogenesis of pulmonary fibrosis and may serve as a molecular target for therapeutic intervention to suppress fibrosis. PMID- 15133033 TI - The cross-Rho'ds of cell-cell adhesion. PMID- 15133034 TI - Evaluation of epigallocatechin gallate and related plant polyphenols as inhibitors of the FabG and FabI reductases of bacterial type II fatty-acid synthase. AB - Epigallocatechin gallate (EGCG) is the major component of green tea extracts and possesses antibacterial, antiviral, and antitumor activity. Our study focused on validating the inhibition of the bacterial type II fatty acid synthesis system as a mechanism for the antibacterial effects of EGCG and related plant polyphenols. EGCG and the related tea catechins potently inhibited both the FabG and FabI reductase steps in the fatty acid elongation cycle with IC(50) values between 5 and 15 microm. The presence of the galloyl moiety was essential for activity, and EGCG was a competitive inhibitor of FabI and a mixed type inhibitor of FabG demonstrating that EGCG interfered with cofactor binding in both enzymes. EGCG inhibited acetate incorporation into fatty acids in vivo, although it was much less potent than thiolactomycin, a validated fatty acid synthesis inhibitor, and overexpression of FabG, FabI, or both did not confer resistance. A panel of other plant polyphenols was screened for FabG/FabI inhibition and antibacterial activity. Most of these inhibited both reductase steps, possessed antibacterial activity, and inhibited cellular fatty acid synthesis. The ability of the plant secondary metabolites to interfere with the activity of multiple NAD(P)-dependent cellular processes must be taken into account when assessing the specificity of their effects. PMID- 15133035 TI - Structure of human MTH1, a Nudix family hydrolase that selectively degrades oxidized purine nucleoside triphosphates. AB - Oxygen radicals generated through normal cellular respiration processes can cause mutations in genomic and mitochondrial DNA. Human MTH1 hydrolyzes oxidized purine nucleoside triphosphates, such as 8-oxo-dGTP and 2-hydroxy-dATP, to monophosphates, thereby preventing the misincorporation of these oxidized nucleotides during replication. Here we present the solution structure of MTH1 solved by multidimensional heteronuclear NMR spectroscopy. The protein adopts a fold similar to that of Escherichia coli MutT, despite the low sequence similarity between these proteins outside the conserved Nudix motif. The substrate-binding pocket of MTH1, deduced from chemical shift perturbation experiments, is located at essentially the same position as in MutT; however, a pocket-forming helix is largely displaced in MTH1 (approximately 9 A) such that the shape of the pocket differs between the two proteins. Detailed analysis of the pocket-forming residues enabled us to identify Asn33 as one of the key residues in MTH1 for discriminating the oxidized form of purine, and mutation of this residue modifies the substrate specificity. We also show that MTH1 catalyzes hydrolysis of 8-oxo-dGTP through nucleophilic substitution of water at the beta phosphate. PMID- 15133036 TI - Phosphorylation of mouse glutamine-fructose-6-phosphate amidotransferase 2 (GFAT2) by cAMP-dependent protein kinase increases the enzyme activity. AB - A protein encoded by a new gene with approximately 75% homology to glutamine fructose-6-phosphate amidotransferase (GFAT) was termed GFAT2 on the basis of this similarity. The mouse GFAT2 cDNA was cloned, and the protein was expressed with either an N-terminal glutathione S-transferase or His tag. The purified protein expressed in mammalian cells had GFAT activity. The Km values for the two substrates of reaction, fructose 6-phosphate and glutamine, were determined to be 0.8 mm for fructose 6-phosphate and 1.2 mm for glutamine, which are within the ranges determined for GFAT1. The protein sequence around the serine 202 of GFAT2 was conserved to the serine 205 of GFAT1, whereas the serine at 235 in GFAT1 was not present in GFAT2. Previously we showed that phosphorylation of serine 205 in GFAT1 by the catalytic subunit of cAMP-dependent protein kinase (PKA) inhibits its activity. Like GFAT1, GFAT2 was phosphorylated by PKA, but GFAT2 activity increased approximately 2.2-fold by this modification. When serine 202 of GFAT2 was mutated to an alanine, the enzyme not only became resistant to phosphorylation, but also the increase in activity in response to PKA also was blocked. These results indicated that the phosphorylation of serine 202 was necessary and sufficient for these alterations by PKA. GFAT2 was modestly inhibited (15%) by UDP-GlcNAc but not through detectable O-glycosylation. GFAT2 is, therefore, an isoenzyme of GFAT1, but its regulation by cAMP is the opposite, allowing differential regulation of the hexosamine pathway in specialized tissues. PMID- 15133037 TI - The major vault protein is a novel substrate for the tyrosine phosphatase SHP-2 and scaffold protein in epidermal growth factor signaling. AB - The catalytic activity of the Src homology 2 (SH2) domain-containing tyrosine phosphatase, SHP-2, is required for virtually all of its signaling effects. Elucidating the molecular mechanisms of SHP-2 signaling, therefore, rests upon the identification of its target substrates. In this report, we have used SHP-2 substrate-trapping mutants to identify the major vault protein (MVP) as a putative SHP-2 substrate. MVP is the predominant component of vaults that are cytoplasmic ribonucleoprotein complexes of unknown function. We show that MVP is dephosphorylated by SHP-2 in vitro and it forms an enzyme-substrate complex with SHP-2 in vivo. In response to epidermal growth factor (EGF), SHP-2 associates via its SH2 domains with tyrosyl-phosphorylated MVP. MVP also interacts with the activated form of the extracellular-regulated kinases (Erks) in response to EGF and a constitutive complex between tyrosyl-phosphorylated MVP, SHP-2, and the Erks was detected in MCF-7 breast cancer cells. Using MVP-deficient fibroblasts, we demonstrate that MVP cooperates with Ras for optimal EGF-induced Elk-1 activation and is required for cell survival. We propose that MVP functions as a novel scaffold protein for both SHP-2 and Erk. The regulation of MVP tyrosyl phosphorylation by SHP-2 may play an important role in cell survival signaling. PMID- 15133038 TI - Growth differentiation factor 9 regulates expression of the bone morphogenetic protein antagonist gremlin. AB - Growth differentiation factor 9 (GDF9) is an oocyte-expressed member of the transforming growth factor beta (TGF-beta) superfamily and is required for normal ovarian follicle development and female fertility. GDF9 acts as a paracrine factor and affects granulosa cell physiology. Only a few genes regulated by GDF9 are known. Our microarray analysis has identified gremlin as one of the genes up regulated by GDF9 in cultures of granulosa cells. Gremlin is a known member of the DAN family of bone morphogenetic protein (BMP) antagonists, but its expression and function in the ovary are unknown. We have investigated the regulation of gremlin in mouse granulosa cells by GDF9 as well as other members of the TGF-beta superfamily. GDF9 and BMP4 induce gremlin, but TGF-beta does not. In addition, in cultures of granulosa cells, gremlin negatively regulates BMP4 signaling but not GDF9 activity. The expression of gremlin in the ovary was also examined by in situ hybridization. A distinct change in gremlin mRNA compartmentalization occurs during follicle development and ovulation, indicating a highly regulated expression pattern during folliculogenesis. We propose that gremlin modulates the cross-talk between GDF9 and BMP signaling that is necessary during follicle development because both ligands use components of the same signaling pathway. PMID- 15133039 TI - Identification of an androgen response element in intron 8 of the sterol regulatory element-binding protein cleavage-activating protein gene allowing direct regulation by the androgen receptor. AB - Sterol regulatory element-binding proteins (SREBPs) are transcription regulators that play a pivotal role in intracellular lipid homeostasis. They are synthesized as inactive precursor proteins in the endoplasmic reticulum, where they are retained by SREBP cleavage-activating protein (SCAP), a sterol sensing protein that in turn is linked to a retention protein complex. Low intracellular sterol concentrations weaken the interaction of SCAP with its retention proteins and allow translocation of the SREBP.SCAP complex to the Golgi compartment where SREBP is proteolytically cleaved and activated. Previous studies on the mechanisms by which androgens provoke a coordinated activation of lipogenic pathways in prostate cancer cells have suggested an alternative pathway of activation in which androgens increase the expression of SCAP and favor translocation of the SREBP.SCAP complex to the Golgi apparatus by disturbing the balance between SCAP and its retention proteins. Here we show that the SCAP gene contains an androgen-responsive region located in intron 8. This region interacts directly with the androgen receptor and confers androgen responsiveness to promoter-reporter constructs transfected in LNCaP cells. It contains a noncanonical androgen response element GGAAGAaaaTGTACC that interacts not only with the androgen receptor but also with the glucocorticoid receptor and that also confers glucocorticoid responsiveness. The identification of a steroid response element in intron 8 of the SCAP gene further supports the contention that SCAP is a direct target for steroid hormone action. PMID- 15133040 TI - Prefibrillar amyloid protein aggregates share common features of cytotoxicity. AB - The intracellular free Ca(2+) concentration and redox status of murine fibroblasts exposed to prefibrillar aggregates of the HypF N-terminal domain have been investigated in vitro and in vivo using a range of fluorescent probes. Aggregate entrance into the cytoplasm is followed by an early rise of reactive oxygen species and free Ca(2+) levels and eventually by cell death. Such changes correlate directly with the viability of the cells and are not observed when cell are cultured in the presence of reducing agents or in Ca(2+)-free media. In addition, moderate cell stress following exposure to the aggregates was found to be fully reversible. The results show that the cytotoxicity of prefibrillar aggregates of HypF-N, a protein not associated with clinical disease, has the same fundamental origin as that produced by similar types of aggregates of proteins linked with specific amyloidoses. These findings suggest that misfolded proteinaceous aggregates stimulate generic cellular responses as a result of the exposure of regions of the structure (such as hydrophobic residues and the polypeptide main chain) that are buried in the normally folded proteins. They also support the idea that a higher number of degenerative pathologies than previously known might be considered as protein deposition diseases. PMID- 15133041 TI - Cti6 is an Rpd3-Sin3 histone deacetylase-associated protein required for growth under iron-limiting conditions in Saccharomyces cerevisiae. AB - Iron and copper are redox active metals essential for life. In the budding yeast Saccharomyces cerevisiae, expression of iron and copper genes involved in metal acquisition and utilization is tightly regulated at the transcriptional level. In addition iron and copper metabolism are inextricably linked because of the dependence on copper as a co-factor for iron uptake or mobilization. To further identify genes that function in iron and copper homeostasis, we screened for novel yeast mutants defective for iron limiting growth and thereby identified the CTI6 gene. Cti6 is a PHD finger-containing protein that has been shown to participate in the interaction of the Ssn6-Tup1 co-repressor with the Gcn5 containing SAGA chromatin-remodeling complex. In this report we show that CTI6 mRNA levels are increased under iron-limiting conditions, and that cti6 mutants display a growth defect under conditions of iron deprivation. Furthermore, we demonstrate that Cti6 is a nuclear protein that functionally associates with the Rpd3-Sin3 histone deacetylase complex involved in transcriptional repression. Cti6 demonstrates Rpd3-dependent transcriptional repression, and cti6 mutants exhibit an enhanced silencing of telomeric, rDNA and HMR loci, similar to mutants in genes encoding other Rpd3-Sin3-associated proteins. Microarray experiments with cti6 mutants grown under iron-limiting conditions show a down-regulation of telomeric genes and an up-regulation of Aft1 and Tup1 target genes involved in iron and oxygen regulation. Taken together, these data suggest a specific role for Cti6 in the regulation of gene expression under conditions of iron limitation. PMID- 15133042 TI - Phosphatidylinositol 3-kinase and frabin mediate Cryptosporidium parvum cellular invasion via activation of Cdc42. AB - Cryptosporidium parvum invades target epithelia via a mechanism that involves host cell actin reorganization. We previously demonstrated that C. parvum activates the Cdc42/neural Wiskott-Aldrich syndrome protein network in host cells resulting in actin remodeling at the host cell-parasite interface, thus facilitating C. parvum cellular invasion. Here, we tested the role of phosphatidylinositol 3-kinase (PI3K) and frabin, a guanine nucleotide exchange factor specific for Cdc42 in the activation of Cdc42 during C. parvum infection of biliary epithelial cells. We found that C. parvum infection of cultured human biliary epithelial cells induced the accumulation of PI3K at the host cell parasite interface and resulted in the activation of PI3K in infected cells. Frabin also was recruited to the host cell-parasite interface, a process inhibited by two PI3K inhibitors, wortmannin and LY294002. The cellular expression of either a dominant negative mutant of PI3K (PI3K-Deltap85) or functionally deficient mutants of frabin inhibited C. parvum-induced Cdc42 accumulation at the host cell-parasite interface. Moreover, LY294002 abolished C. parvum-induced Cdc42 activation in infected cells. Inhibition of PI3K by cellular overexpression of PI3K-Deltap85 or by wortmannin or LY294002, as well as inhibition of frabin by various functionally deficient mutants, decreased C. parvum-induced actin accumulation and inhibited C. parvum cellular invasion. In contrast, the overexpression of the p85 subunit of PI3K promoted C. parvum invasion. Our data suggest that an important component of the complex process of C. parvum invasion of target epithelia results from the ability of the organism to trigger host cell PI3K/frabin signaling to activate the Cdc42 pathway, resulting in host cell actin remodeling at the host cell-parasite interface. PMID- 15133043 TI - Functional properties of the herpes simplex virus type I origin-binding protein are controlled by precise interactions with the activated form of the origin of DNA replication. AB - The herpes simplex virus, type I origin-binding protein, OBP, is a superfamily II DNA helicase encoded by the UL9 gene. OBP binds in a sequence-specific and cooperative way to the viral origin of replication oriS. OBP may unwind partially and introduce a hairpin into the double-stranded origin of replication. The formation of the novel conformation referred to as oriS* also requires the single stranded DNA-binding protein, ICP8, and ATP hydrolysis. OBP forms a stable complex with oriS*. The hairpin in oriS* provides a site for sequence-specific attachment, and a single-stranded region triggers ATP hydrolysis. Here we use Escherichia coli exonuclease I to map the binding of the C-terminal domain of OBP to the hairpin and the helicase domains to the single-stranded tail. The helicase domains cover a stretch of 23 nucleotides of single-stranded DNA. Using streptavidin-coated magnetic beads, we show that OBP may bind two copies of double-stranded DNA (one biotin-labeled and the other one radioactively labeled) but only one copy of oriS*. It is the length of the single-stranded tail that determines the stoichiometry of OBP.DNA complexes. OBP interacts with the bases of the single-stranded tail, and ATP hydrolysis is triggered by position-specific interactions between OBP and bases in the single-stranded tail of oriS*. PMID- 15133044 TI - The effects of HIV-1 Nef on CD4 surface expression and viral infectivity in lymphoid cells are independent of rafts. AB - The HIV-1 Nef protein is a critical virulence factor that exerts multiple effects during viral replication. Nef modulates surface expression of various cellular proteins including CD4 and MHC-I, enhances viral infectivity, and affects signal transduction pathways. Nef has been shown to partially associate with rafts, where it can prime T cells for activation. The contribution of rafts during Nef induced CD4 down-regulation and enhancement of viral replication remains poorly understood. We show here that Nef does not modify the palmitoylation state of CD4 or its partition within rafts. Moreover, CD4 mutants lacking palmitoylation or unable to associate with rafts are efficiently down-regulated by Nef. In HIV infected cells, viral assembly and budding occurs from rafts, and Nef has been suggested to increase this process. However, using T cells acutely infected with wild-type or nef-deleted HIV, we did not observe any impact of Nef on raft segregation of viral structural proteins. We have also designed a palmitoylated mutant of Nef (NefG3C), which significantly accumulates in rafts. Interestingly, the efficiency of NefG3C to down-regulate CD4 and MHC-I, and to promote viral replication was not increased when compared with the wild-type protein. Altogether, these results strongly suggest that rafts are not a key element involved in the effects of Nef on trafficking of cellular proteins and on viral replication. PMID- 15133046 TI - PrPSc binding antibodies are potent inhibitors of prion replication in cell lines. AB - Conversion of the cellular alpha-helical prion protein (PrP(C)) into a disease associated isoform (PrP(Sc)) is central to the pathogenesis of prion diseases. Molecules targeting either normal or disease-associated isoforms may be of therapeutic interest, and the antibodies binding PrP(C) have been shown to inhibit prion accumulation in vitro. Here we investigate whether antibodies that additionally target disease-associated isoforms such as PrP(Sc) inhibit prion replication in ovine PrP-inducible scrapie-infected Rov cells. We conclude from these experiments that antibodies exclusively binding PrP(C) were relatively inefficient inhibitors of ScRov cell PrP(Sc) accumulation compared with antibodies that additionally targeted disease-associated PrP isoforms. Although the mechanism by which these monoclonal antibodies inhibit prion replication is unclear, some of the data suggest that antibodies might actively increase PrP(Sc) turnover. Thus antibodies that bind to both normal and disease-associated isoforms represent very promising anti-prion agents. PMID- 15133045 TI - Molecular basis of the high insecticidal potency of scorpion alpha-toxins. AB - Scorpion alpha-toxins are similar in their mode of action and three-dimensional structure but differ considerably in affinity for various voltage-gated sodium channels (NaChs). To clarify the molecular basis of the high potency of the alpha toxin LqhalphaIT (from Leiurus quinquestriatus hebraeus) for insect NaChs, we identified by mutagenesis the key residues important for activity. We have found that the functional surface is composed of two distinct domains: a conserved "Core-domain" formed by residues of the loops connecting the secondary structure elements of the molecule core and a variable "NC-domain" formed by a five-residue turn (residues 8-12) and a C-terminal segment (residues 56-64). We further analyzed the role of these domains in toxin activity on insects by their stepwise construction onto the scaffold of the anti-mammalian alpha-toxin, Aah2 (from Androctonus australis hector). The chimera harboring both domains, Aah2(LqhalphaIT(face)), was as active to insects as LqhalphaIT. Structure determination of Aah2(LqhalphaIT(face)) by x-ray crystallography revealed that the NC-domain deviates from that of Aah2 and forms an extended protrusion off the molecule core as appears in LqhalphaIT. Notably, such a protrusion is observed in all alpha-toxins active on insects. Altogether, the division of the functional surface into two domains and the unique configuration of the NC-domain illuminate the molecular basis of alpha-toxin specificity for insects and suggest a putative binding mechanism to insect NaChs. PMID- 15133047 TI - A molecular handoff between bacteriophage T7 DNA primase and T7 DNA polymerase initiates DNA synthesis. AB - The T7 DNA primase synthesizes tetraribonucleotides that prime DNA synthesis by T7 DNA polymerase but only on the condition that the primase stabilizes the primed DNA template in the polymerase active site. We used NMR experiments and alanine scanning mutagenesis to identify residues in the zinc binding domain of T7 primase that engage the primed DNA template to initiate DNA synthesis by T7 DNA polymerase. These residues cover one face of the zinc binding domain and include a number of aromatic amino acids that are conserved in bacteriophage primases. The phage T7 single-stranded DNA-binding protein gp2.5 specifically interfered with the utilization of tetraribonucleotide primers by interacting with T7 DNA polymerase and preventing a productive interaction with the primed template. We propose that the opposing effects of gp2.5 and T7 primase on the initiation of DNA synthesis reflect a sequence of mutually exclusive interactions that occur during the recycling of the polymerase on the lagging strand of the replication fork. PMID- 15133048 TI - Acute formation of protease-resistant prion protein does not always lead to persistent scrapie infection in vitro. AB - Transmissible spongiform encephalopathies are accompanied by the accumulation of a pathologic isoform of a host-encoded protein, termed prion protein (PrP). Despite the widespread distribution of the cellular isoform of PrP (protease sensitive PrP; PrP-sen), the disease-associated isoform (protease-resistant PrP; PrP-res) appears to be primarily restricted to cells of the nervous and lymphoreticular systems. In order to study why scrapie infection appears to be restricted to certain cells, we followed acute and persistent PrP-res formation upon exposure of cells to different scrapie agents. We found that, independent of the cell type and scrapie strain, initial PrP-res formation occurred rapidly in cells. However, sustained generation of PrP-res and persistent infection did not necessarily follow acute PrP-res formation. Persistent PrP-res formation and scrapie infection was restricted to one cell line inoculated with the mouse scrapie strain 22L. In contrast to cells that did not become scrapie-infected, the level of PrP-res in the 22L-infected cells rapidly increased in the absence of a concomitant increase in the number of PrP-res-producing cells. Furthermore, the protein banding pattern of PrP-res in these cells changed over time as the cells became chronically infected. Thus, our results suggest that the events leading to the initial formation of PrP-res may differ from those required for sustained PrP-res formation and infection. This may, at least in part, explain the observation that not all PrP-sen-expressing cells appear to support transmissible spongiform encephalopathy agent replication. PMID- 15133049 TI - NMR structure of the N-terminal domain of SUMO ligase PIAS1 and its interaction with tumor suppressor p53 and A/T-rich DNA oligomers. AB - A member of the PIAS (protein inhibitor of activated STAT) family of proteins, PIAS1, have been reported to serve as an E3-type SUMO ligase for tumor suppressor p53 and its own. It also was proposed that the N-terminal domain of PIAS1 interacts with DNA as well as p53. Extensive biochemical studies have been devoted recently to understand sumoylations and its biological implications, whereas the structural aspects of the PIAS family and the mechanism of its interactions with various factors are less well known to date. In this study, the three-dimensional structure of the N-terminal domain (residues 1-65) of SUMO ligase PIAS1 was determined by NMR spectroscopy. The structure revealed a unique four-helix bundle with a topology of an up-down-extended loop-down-up, a part of which the helix-extended loop-helix represented the SAP (SAF-A/B, Acinus, and PIAS) motif. Thus, this N-terminal domain may be referred to as a four-helix SAP domain. The glutathione S-transferase pull-down assay demonstrated that this domain possesses a binding ability to tumor suppressor p53, a target protein for sumoylation by PIAS1, whereas gel mobility assays showed that it has a strong affinity toward A/T-rich DNA. An NMR analysis of the four-helix SAP domain complexed with the 16-bp-long DNA demonstrated that one end of the four-helix bundle is the binding site and may fit into the minor groove of DNA. The three dimensional structure and its binding duality are discussed in conjunction with the biological functions of PIAS1 as a SUMO ligase. PMID- 15133051 TI - Hydraulic conductance and rootstock effects in grafted vines of kiwifruit. AB - Whole-plant hydraulic conductance, shoot growth, and leaf photosynthetic properties were measured on kiwifruit vines with four clonal rootstocks to examine the relationship between plant hydraulic conductance and leaf stomatal conductance (gs) and to test the hypothesis that reduced hydraulic conductance can provide an explanation for reductions in plant vigour caused by rootstocks. The rootstocks were selected from four species of Actinidia and grafted with Actinidia chinensis var. chinensis 'Hort16A' (yellow kiwifruit) as the scion. Total leaf area of the scion on the least vigorous Actinidia rootstock, A. kolomikta, was 25% of the most vigorous, A. hemsleyana. Based on shoot growth and leaf area, the selections of A. kolomikta and A. polygama are low-vigour rootstocks, and A. macrosperma and A. hemsleyana are high-vigour rootstocks for A. chinensis. Whole-plant hydraulic conductance, the ratio of xylem sap flux to xylem water potential, was lower in the low-vigour rootstocks, reflecting their smaller size. However, leaf-area-specific conductance (Kl) and gs were both higher in the low-vigour rootstocks, the opposite of the expected pattern. Differences in Kl were found in the compartment from the roots to the scion stem, with no difference between rootstocks in the conductance of stems or leaves of the scion. There was no evidence that the graft union caused a significant reduction in hydraulic conductance of vines with low-vigour rootstocks. Leaf photosynthetic capacity did not vary between rootstocks, but photosynthesis and carbon isotope discrimination (Delta13C) under ambient conditions were higher in the low-vigour rootstocks because gs was higher. gs and Delta13C were positively correlated with Kl, although the mechanism for this relationship was not based on stomatal regulation of a similar xylem water potential because water potential varied between rootstocks. For Actinidia rootstocks, changes in Kl do not provide a direct explanation for changes in vigour of the scion. However, depending on the rootstock in question, changes in hydraulic conductance, biomass partitioning, and crown structure are involved in the response. PMID- 15133050 TI - Molecular analysis and control of cysteine biosynthesis: integration of nitrogen and sulphur metabolism. AB - Since cysteine is the first committed molecule in plant metabolism containing both sulphur and nitrogen, the regulation of its biosynthesis is critically important. Cysteine itself is required for the production of an abundance of key metabolites in diverse pathways. Plants alter their metabolism to compensate for sulphur and nitrogen deficiencies as best as they can, but limitations in either nutrient not only curb a plant's ability to synthesize cysteine, but also restrict protein synthesis. Nutrients such as nitrate and sulphate (and carbon) act as signals; they trigger molecular mechanisms that modify biosynthetic pathways and thereby have a profound impact on metabolite fluxes. Cysteine biosynthesis is modified by regulators acting at the site of uptake and throughout the plant system. Recent data point to the existence of nutrient specific signal transduction pathways that relay information about external and internal nutrient concentrations, resulting in alterations to cysteine biosynthesis. Progress in this field has led to the cloning of genes that play pivotal roles in nutrient-induced changes in cysteine formation. PMID- 15133052 TI - Boron nutrition of cultured tobacco BY-2 cells. IV. Genes induced under low boron supply. AB - Genes whose expression was up-regulated in low boron (B)-acclimated tobacco BY-2 (Nicotiana tabacum L. cv. Bright Yellow 2) cells, which had been selected under a low supply of B, were screened by the cDNA differential subtraction method. Thirteen genes were identified, including early salicylate-inducible glucosyltransferase, glutamine synthetase, glutathione S-transferase, and a pathogenesis-related protein, which might constitute a rescue system for oxidative damage. This indicates that B deficiency might impose cellular redox imbalance on the cells. Two of the 13 genes were induced within 30 min of B removal in the parent cells, indicating fast signal transfer from the cell walls to the cytoplasm. PMID- 15133053 TI - Responses of primary and secondary metabolism to sugar accumulation revealed by microarray expression analysis of the Arabidopsis mutant, pho3. AB - The Arabidopsis mutant pho3 accumulates sucrose and other carbohydrates to high levels, providing a means of investigating the genomic response to sucrose accumulation using microarray analysis. Wild-type and mutant plants were grown in soil to the mature rosette stage for the analysis of gene expression using the Affymetrix ATH1 chip, containing more than 22,500 probe sets. Small, but significant, decreases were observed in the expression of many genes encoding enzymes and regulatory proteins involved in primary carbon assimilation, suggesting that, in mature leaves of Arabidopsis, there is limited feedback regulation on gene expression by sugars. The study revealed a striking increase in the expression of the plastid glucose 6-phosphate/phosphate translocator, characteristically expressed only in heterotrophic tissues. This indicated a change in the nature of metabolite exchange between the plastid and the cytosol in the pho3 mutant. The expression of enzymes of starch synthesis also increased significantly. Very large increases were observed in the expression of transcription factors and enzymes involved in anthocyanin biosynthesis. This finding reinforces the emerging picture of an important role for primary metabolism in regulating secondary metabolism. PMID- 15133054 TI - Isolation of cucumber CsARF cDNAs and expression of the corresponding mRNAs during gravity-regulated morphogenesis of cucumber seedlings. AB - Cucumber seedlings show positive gravitropism and bend in the transition zone between the hypocotyl and the root. The peg, a specialized protuberance, develops on the concave side of the bending transition zone. Auxin and the mRNA of an auxin-inducible gene (CsIAA1) isolated from cucumber are differentially accumulated across the transition zone during the gravity-regulated peg formation. In this study, five cDNAs of Auxin Response Factors (ARFs) from cucumber were isolated and their mRNA accumulation was compared with that of CsIAA1. The tissue specificity of CsARF2 mRNA accumulation was similar to that of CsIAA1. Because the structural character of CsARF2 predicts that it is a transcriptional activator, CsARF2 may be involved in the activation of CsIAA1 transcription, which plays a role in gravity-regulated peg formation. Neither gravity nor auxin affected mRNA accumulation of five CsARFs including CsARF2, suggesting that CsARF2 may be regulated at a post-transcriptional level to induce the asymmetric expression of the CsIAA1 gene in response to gravistimulation and auxin in cucumber seedlings. PMID- 15133055 TI - Combining thermal and visible imagery for estimating canopy temperature and identifying plant stress. AB - Thermal imaging is a potential tool for estimating plant temperature, which can be used as an indicator of stomatal closure and water deficit stress. In this study, a new method for processing and analysing thermal images was developed. By using remote sensing software, the information from thermal and visible images was combined, the images were classified to identify leaf area and sunlit and shaded parts of the canopy, and the temperature statistics for specific canopy components were calculated. The method was applied to data from a greenhouse water-stress experiment of Vicia faba L. and to field data for Vitis vinifera L. Vaseline-covered and water-sprayed plants were used as dry and wet references, respectively, and two thermal indices, based on temperature differences between the canopy and reference surfaces, were calculated for single Vicia faba plants. The thermal indices were compared with measured stomatal conductance. The temperature distributions of sunlit and shaded leaf area of Vitis vinifera canopies from natural rainfall and irrigation treatments were compared. The present method provides two major improvements compared with earlier methods for calculating thermal indices. First, it allows more accurate estimation of the indices, which are consequently more closely related to stomatal conductance. Second, it gives more accurate estimates of the temperature distribution of the shaded and sunlit parts of canopy, and, unlike the earlier methods, makes it possible to quantify the relationship between temperature variation and stomatal conductance. PMID- 15133056 TI - Regulatory levels for the transport of ammonium in plant roots. AB - Ammonium is an attractive nitrogen form for root uptake due to its permanent availability and the reduced state of the nitrogen. On the other hand, ammonium fluxes are difficult to control because ammonium represents an equilibrium between NH4+ and NH3, which are two N forms with different membrane permeabilities. There is increasing evidence that AMT-type ammonium transporters represent the major entry pathways for root uptake of NH4+. Since excess uptake of ammonium might cause toxicity and since ammonium is also released from catabolic processes within the cell, ammonium uptake across the root plasma membrane has to be tightly regulated. To take over a function in cellular ammonium homeostasis, various AMT transporters are synthesized that differ in their biochemical properties, their localization, and in their regulation at the transcriptional level. At the same time, AMT-driven transport is subject to control by the nitrogen status of a local root portion as well as of the whole plant. In this review, the focus is on the different levels at which AMT dependent ammonium uptake is regulated and the gaps in current knowledge are highlighted. PMID- 15133057 TI - Tomato fruit cuticular waxes and their effects on transpiration barrier properties: functional characterization of a mutant deficient in a very-long chain fatty acid beta-ketoacyl-CoA synthase. AB - Cuticular waxes play a pivotal role in limiting transpirational water loss across the plant surface. The correlation between the chemical composition of the cuticular waxes and their function as a transpiration barrier is still unclear. In the present study, intact tomato fruits (Lycopersicon esculentum) are used, due to their astomatous surface, as a novel integrative approach to investigate this composition- function relationship: wax amounts and compositions of tomato were manipulated before measuring unbiased cuticular transpiration. First, successive mechanical and extractive wax-removal steps allowed the selective modification of epi- and intracuticular wax layers. The epicuticular film consisted exclusively of very-long-chain aliphatics, while the intracuticular compartment contained large quantities of pentacyclic triterpenoids as well. Second, applying reverse genetic techniques, a loss-of-function mutation with a transposon insertion in a very-long-chain fatty acid elongase beta-ketoacyl-CoA synthase was isolated and characterized. Mutant leaf and fruit waxes were deficient in n-alkanes and aldehydes with chain lengths beyond C30, while shorter chains and branched hydrocarbons were not affected. The mutant fruit wax also showed a significant increase in intracuticular triterpenoids. Removal of the epicuticular wax layer, accounting for one-third of the total wax coverage on wild-type fruits, had only moderate effects on transpiration. By contrast, reduction of the intracuticular aliphatics in the mutant to approximately 50% caused a 4-fold increase in permeability. Hence, the main portion of the transpiration barrier is located in the intracuticular wax layer, largely determined by the aliphatic constituents, but modified by the presence of triterpenoids, whereas epicuticular aliphatics play a minor role. PMID- 15133058 TI - Expression analysis of a chicory fructan 1-exohydrolase gene reveals complex regulation by cold. AB - The gene for a recently identified cDNA, 1-FEH IIa, encoding a fructan 1 exohydrolase was isolated and cloned from Cichorium intybus and a 1149 bp promoter fragment was characterized. An analysis of the genomic 1-FEH IIa sequence indicated that the gene (FEHIIa) consists of six introns and seven exons, which is similar to plant invertase genes. Like invertase genes, FEHIIa also contains the 9 nt mini-exon encoding the tripeptide DPN. A database search for cis-acting response elements within its promoter identified multiple elements that appear to have relevance to cold-induced expression of the gene in field grown roots. Promoter analysis by transient expression assay demonstrated that the FEHIIa gene promoter is highly expressed in etiolated Cichorium leaves and cold-stored roots, which correlated well with the high level expression detected by RNA blot analysis. Cold also enhanced FEHIIa reporter gene expression in green leaves, however, the reporter gene activity was much lower compared with similar induction experiments in etiolated leaves. Promoter deletion analysis demonstrated the presence of potential cold-responsive ABRE and/or CRT/DRE elements in the -22 to -172 region, while regions -933 to -717 and -493 to -278 contain elements that can down-regulate expression at the conditions used. Characterization of the FEHIIa promoter may provide tools to study cold-induced expression and to increase freezing tolerance in agricultural crops. PMID- 15133059 TI - The slow reversibility of photosystem II thermal energy dissipation on transfer from high to low light may cause large losses in carbon gain by crop canopies: a theoretical analysis. AB - Regulated thermal dissipation of absorbed light energy within the photosystem II antenna system helps protect photosystem II from damage in excess light. This reversible photoprotective process decreases the maximum quantum yield of photosystem II (Fv)/Fm) and CO2 assimilation (phiCO2), and decreases the convexity of the non-rectangular hyperbola describing the response of leaf CO2 assimilation to photon flux (theta). At high light, a decrease in phiCO2 has minimal impact on carbon gain, while high thermal energy dissipation protects PSII against oxidative damage. Light in leaf canopies in the field is continually fluctuating and a finite period of time is required for recovery of phiCO2 and when light drops below excess levels. Low phiCO2) and can limit the rate of photosynthetic carbon assimilation on transfer to low light, an effect prolonged by low temperature. What is the cost of this delayed reversal of thermal energy dissipation and phiCO2 recovery to potential CO2 uptake by a canopy in the field? To address this question a reverse ray-tracing algorithm for predicting the light dynamics of 120 randomly selected individual points in a model canopy was used to describe the discontinuity and heterogeneity of light flux within the canopy. Because photoprotection is at the level of the cell, not the leaf, light was simulated for small points of 10(4) micro m rather than as an average for a leaf. The predicted light dynamics were combined with empirical equations simulating the dynamics of the light-dependent decrease and recovery of phiCO2 and and their effects on the integrated daily canopy carbon uptake (A'c). The simulation was for a model canopy of leaf area index 3 with random inclination and orientation of foliage, on a clear sky day (latitude 44 degrees N, 120th day of the year). The delay in recovery of photoprotection was predicted to decrease A'c by 17% at 30 degrees C and 32% at 10 degrees C for a chilling-susceptible species, and by 12.8% at 30 degrees C and 24% at 10 degrees C for a chilling-tolerant species. These predictions suggest that the selection, or engineering, of genotypes capable of more rapid recovery from the photoprotected state would substantially increase carbon uptake by crop canopies in the field. PMID- 15133060 TI - Biochemical characterization of protein complexes from the Helicobacter pylori protein interaction map: strategies for complex formation and evidence for novel interactions within type IV secretion systems. AB - We have investigated a large set of interactions from the Helicobacter pylori protein interaction map previously identified by high-throughput yeast two-hybrid (htY2H)-based methods. This study had two aims: i) to validate htY2H as a source of protein-protein interaction complexes for high-throughput biochemical and structural studies of the H. pylori interactome; and ii) to validate biochemically interactions shown by htY2H to involve components of the H. pylori type IV secretion systems. Thus, 17 interactions involving 31 proteins and protein fragments were studied, and a general strategy was designed to produce protein-interacting partners for biochemical and structural characterization. We show that htY2H is a valid source of protein-protein complexes for high throughput proteome-scale characterization of the H. pylori interactome, because 76% of the interactions tested were confirmed biochemically. Of the interactions involving type IV secretion proteins, three could be confirmed. One interaction is between two components of the type IV secretion apparatus, ComB10 and ComB4, which are VirB10 and VirB4 homologs, respectively. Another interaction is between a type IV component (HP0525, a VirB11 homolog) and a non-type IV secretion protein (HP01451), indicating that proteins other than the core VirB (1-11)-VirD4 proteins may play a role in type IV secretion. Finally, a third interaction was biochemically confirmed between CagA, a virulence factor secreted by the type IV secretion system encoded by the Cag pathogenicity island, and a non-type IV secretion protein, HP0496. PMID- 15133061 TI - Exposure to cAMP and beta-adrenergic stimulation recruits Ca(V)3 T-type channels in rat chromaffin cells through Epac cAMP-receptor proteins. AB - T-type channels are expressed weakly or not at all in adult rat chromaffin cells (RCCs) and there is contrasting evidence as to whether they play a functional role in catecholamine secretion. Here we show that 3-5 days after application of pCPT-cAMP, most RCCs grown in serum-free medium expressed a high density of low voltage-activated T-type channels without altering the expression and characteristics of high-voltage-activated channels. The density of cAMP-recruited T-type channels increased with time and displayed the typical biophysical and pharmacological properties of low-voltage-activated Ca(2+) channels: (1) steep voltage-dependent activation from -50 mV in 10 mm Ca(2+), (2) slow deactivation but fast and complete inactivation, (3) full inactivation following short conditioning prepulses to -30 mV, (4) effective block of Ca(2+) influx with 50 microM Ni(2+), (5) comparable permeability to Ca(2+) and Ba(2+), and (6) insensitivity to common Ca(2+) channel antagonists. The action of exogenous pCPT cAMP (200 microM) was prevented by the protein synthesis inhibitor anisomycin and mimicked in most cells by exposure to forskolin and 1-methyl-3-isobutylxanthine (IBMX) or isoprenaline. The protein kinase A (PKA) inhibitor H89 (0.3 microM) and the competitive antagonist of cAMP binding to PKA, Rp-cAMPS, had weak or no effect on the action of pCPT-cAMP. In line with this, the selective Epac agonist 8CPT-2Me-cAMP nicely mimicked the action of pCPT-cAMP and isoprenaline, suggesting the existence of a dominant Epac-dependent recruitment of T-type channels in RCCs that may originate from the activation of beta-adrenoceptors. Stimulation of beta-adrenoceptors occurs autocrinally in RCCs and thus, the neosynthesis of low-voltage-activated channels may represent a new form of 'chromaffin cell plasticity', which contributes, by lowering the threshold of action potential firing, to increasing cell excitability and secretory activity during sustained sympathetic stimulation and/or increased catecholamine circulation. PMID- 15133062 TI - Postnatal microbial colonization programs the hypothalamic-pituitary-adrenal system for stress response in mice. AB - Indigenous microbiota have several beneficial effects on host physiological functions; however, little is known about whether or not postnatal microbial colonization can affect the development of brain plasticity and a subsequent physiological system response. To test the idea that such microbes may affect the development of neural systems that govern the endocrine response to stress, we investigated hypothalamic-pituitary-adrenal (HPA) reaction to stress by comparing germfree (GF), specific pathogen free (SPF) and gnotobiotic mice. Plasma ACTH and corticosterone elevation in response to restraint stress was substantially higher in GF mice than in SPF mice, but not in response to stimulation with ether. Moreover, GF mice also exhibited reduced brain-derived neurotrophic factor expression levels in the cortex and hippocampus relative to SPF mice. The exaggerated HPA stress response by GF mice was reversed by reconstitution with Bifidobacterium infantis. In contrast, monoassociation with enteropathogenic Escherichia coli, but not with its mutant strain devoid of the translocated intimin receptor gene, enhanced the response to stress. Importantly, the enhanced HPA response of GF mice was partly corrected by reconstitution with SPF faeces at an early stage, but not by any reconstitution exerted at a later stage, which therefore indicates that exposure to microbes at an early developmental stage is required for the HPA system to become fully susceptible to inhibitory neural regulation. These results suggest that commensal microbiota can affect the postnatal development of the HPA stress response in mice. PMID- 15133063 TI - Tetrahydrobiopterin improves endothelial dysfunction and vascular oxidative stress in microvessels of intrauterine undernourished rats. AB - In the present study, we investigated the effects of the exogenous application of tetrahydrobiopterin on the endothelium-dependent vasorelaxation and superoxide anion generation in the mesenteric microvessels of intrauterine undernourished rats. In addition, we investigated the presence of peroxynitrite in these rats by evaluation of nitrotyrosine-containing proteins, a stable end-product of peroxynitrite oxidation. For this, female pregnant Wistar rats were fed either normal or 50% of the normal intake diets during the whole gestational period. Male offspring (16 weeks of age) were studied to assess microvascular reactivity, superoxide production using a hydroethidine staining assay, nitric oxide synthase (NOS) activity and nitric oxide (NO) production. Western blot analysis was used to quantify nitrotyrosine-containing proteins and relative multiplex RT-PCR analysis for endothelial NOS (eNOS) mRNA expression. Superfusion with tetrahydrobiopterin significantly decreased superoxide generation and improved vascular function. Intrauterine malnutrition induced a decrement of NOS activity and NO production without affecting the gene expression of eNOS. However, incubation with tetrahydrobiopterin significantly improved NO production after stimulation with acetylcholine or bradykinin in intrauterine undernourished rats. The fact that the nitrotyrosine-containing proteins were increased could, at first sight, suggest that the peroxynitrite is the mediator responsible for the excessive oxidation and depletion of tetrahydrobiopterin. Our study shows that exogenous application of tetrahydrobiopterin leads to a significant improvement of endothelium-dependent vasodilatation, enhanced NO production and decreased superoxide generation in microvessels of intrauterine undernourished rats. Since we found a decrease in NOS activity without an alteration in the gene expression of eNOS, we suggest that impaired NOS-dependent responses of mesenteric arterioles are related to the impairment of tetrahydrobiopterin pathways. PMID- 15133064 TI - Mitochondrial calcium sequestration and protein kinase C cooperate in the regulation of cortical F-actin disassembly and secretion in bovine chromaffin cells. AB - Mitochondria play an important role in the homeostasis of intracellular Ca(2+) and regulate its availability for exocytosis. Inhibitors of mitochondria Ca(2+) uptake such as protonophore CCCP potentiate the secretory response to a depolarizing pulse of K(+). Exposure of cells to agents that directly (cytochalasin D, latrunculin B) or indirectly (PMA) disrupt cortical F-actin networks also potentiate the secretory response to high K(+). The effects of cytochalasin D and CCCP on secretion were additive whereas those of PMA and CCCP were not; this suggests different mechanisms for cytochalasin D and CCCP and a similar mechanism for PMA and CCCP. Mitochondria were the site of action of CCCP, because the potentiation of secretion by CCCP was observed even after depletion of Ca(2+) from the endoplasmic reticulum. CCCP induced a small increase in the cytosolic Ca(2+) concentration ([Ca(2+)](c)) that was not modified by the protein kinase C (PKC) inhibitor chelerythrine. Both CCCP and PMA induced cortical F actin disassembly, an effect abolished by chelerythrine. In addition, rotenone and oligomycin A, two other mitochondrial inhibitors, also evoked cortical F actin disassembly and potentiated secretion; again, these effects were blocked by chelerythrine. CCCP also enhanced the phosphorylation of PKC and myristoylated alanine-rich C kinase substance (MARCKS), and these were also inhibited by chelerythrine. The results suggest that the rapid sequestration of Ca(2+) by mitochondria would protect the cell from an enhanced PKC activation and cortical F-actin disassembly, thereby limiting the magnitude of the secretory response. PMID- 15133067 TI - 'Anaerobic protists': some misconceptions and confusions. PMID- 15133066 TI - Genome update: rRNAs in sequenced microbial genomes. PMID- 15133068 TI - Azotobacter vinelandii: a Pseudomonas in disguise? PMID- 15133065 TI - Maternal nutrient restriction alters gene expression in the ovine fetal heart. AB - Adequate maternal nutrient supply is critical for normal fetal organogenesis. We previously demonstrated that a global 50% nutrient restriction during the first half of gestation causes compensatory growth of both the left and right ventricles of the fetal heart by day 78 of gestation. Thus, it was hypothesized that maternal nutrient restriction significantly altered gene expression in the fetal cardiac left ventricle (LV). Pregnant ewes were randomly grouped into control (100% national research council (NRC) requirements) or nutrient restricted groups (50% NRC requirements) from day 28 to day 78 of gestation, at which time fetal LV were collected. Fetal LV mRNA was used to construct a suppression subtraction cDNA library from which 11 cDNA clones were found by differential dot blot hybridization and virtual Northern analysis to be up regulated by maternal nutrient restriction: caveolin, stathmin, G-1 cyclin, alpha actin, titin, cardiac ankyrin repeat protein (CARP), cardiac-specific RNA helicase activated by MEF2C (CHAMP), endothelial and smooth muscle derived neuropilin (ESDN), prostatic binding protein, NADH dehydrogenase subunit 2, and an unknown protein. Six of these clones (cardiac alpha-actin, cyclin G1, stathmin, NADH dehydrogenase subunit 2, titin and prostatic binding protein) have been linked to cardiac hypertrophy in other species including humans. Of the remaining clones, caveolin, CARP and CHAMP have been shown to inhibit remodelling of hypertrophic tissue. Compensatory growth of fetal LV in response to maternal undernutrition is concluded to be associated with increased transcription of genes related to cardiac hypertrophy, compensatory growth or remodelling. Counter regulatory gene transcription may be increased, in part, as a response to moderating the degree of cardiac remodelling. The short- and long-term consequences of these changes in fetal heart gene expression and induction of specific homeostatic mechanisms in response to maternal undernutrition remain to be determined. PMID- 15133069 TI - A C-terminal deletion mutant of Mycobacterium tuberculosis FtsZ shows fast polymerization in vitro. PMID- 15133070 TI - Screening genomes of Gram-positive bacteria for double-glycine-motif-containing peptides. PMID- 15133071 TI - Anaerobic protists and hidden mitochondria. PMID- 15133073 TI - Production of ammonia by Tritrichomonas foetus and Trichomonas vaginalis. AB - Production of ammonia is difficult to find among the various studies of amino acid metabolism in protozoa. Several studies suggest that catabolism of arginine to ammonium is important for the growth of trichomonads. Trichomonads are amitochondriate zooflagellates that thrive under microaerophilic and anaerobic conditions. The authors were able to detect accumulation of ammonium ions and ammonia in cultures of Tritrichomonas foetus and Trichomonas vaginalis, including those resistant to metronidazole. Ammonium ions and ammonia were detected using the indophenol colorimetric method. Cells incubated overnight under an ambient oxygen gas phase had 0.9 mM soluble ammonium (NH(4)(+) and NH(3)) or a 20 % greater concentration of ammonium relative to sterile growth medium that had been incubated similarly. Production of ammonia itself was confirmed by analysis of a wick that was moistened with sulfuric acid (20 mM) and placed above the liquid in sealed cultures of a strain of Trichomonas vaginalis. The wicks from these cultures captured the equivalent of 0.048 mM volatile ammonia (NH(3)) from the liquid as compared to 0.021 mM volatile ammonia from sterile medium after overnight incubation. Intact trichomonads, 0.7 x 10(6) cells ml(-1) equivalent to 0.7 mg protein ml(-1), incubated in Doran's buffer with or without (1 mM) L arginine produced significant amounts of soluble ammonium (0.07 mM and 0.04 mM, respectively) during 60 min. The results indicate that ammonium ions and the more irritating ammonia are significant metabolites of trichomonads. In addition, based upon end-product amounts, it appears that the rate of arginine metabolism is of the same order of magnitude as that for carbohydrate metabolism by trichomonads. PMID- 15133072 TI - tvcp12: a novel Trichomonas vaginalis cathepsin L-like cysteine proteinase encoding gene. AB - Trichomonas vaginalis is the causative agent of trichomoniasis, one of the most common sexually transmitted diseases in humans. This protozoan has multiple proteinases that are mainly of the cysteine proteinase (CP) type, some of which are known to be involved in the parasite's virulence. Here, a novel T. vaginalis CP-encoding gene, tvcp12, was identified and characterized. tvcp12 is 948 bp long and encodes a predicted 34.4 kDa protein that has the characteristics of the papain-like CP family. TvCP12 does not appear to have a signal peptide, suggesting that this is a cytoplasmic CP. By Southern blot assays, the tvcp12 gene was found as a single copy in the T. vaginalis genome. Remarkably, Northern blot experiments showed a single transcript band of approximately 1.3 kb in the mRNA obtained from parasites grown in low iron conditions and no transcript was observed in the mRNA from parasites grown in high iron conditions. By RT-PCR assays, a 270 bp band was amplified from the cDNA of parasites grown in low iron medium, which was very faint when cDNA from parasites grown in high iron conditions was used. Transcripts of the 3' region obtained in both iron conditions presented differences in their poly(A) tail length. These data suggest that tvcp12 is another gene that is negatively regulated by iron and that the length of the poly(A) tail may be one of the factors involved in the iron modulated protein expression. PMID- 15133074 TI - PCR-based identification of zoonotic isolates of Blastocystis from mammals and birds. AB - The genotype of Blastocystis isolated from humans and animals is highly polymorphic. Therefore, it is important to compare the genotypes of Blastocystis isolates from humans and animals to determine the zoonotic potential of animal isolates. PCR-based genotype classification using known sequence-tagged site (STS) primers allows identification of zoonotic isolates of animal origin. To this end, 51 isolates from monkeys, cattle, pigs, chickens, quails and pheasants were subjected to genotype analysis using seven kinds of STS primers. Out of the 51 isolates, 39 were identified as one of the known genotypes, four showed mixed genotypes, and eight were unknown genotypes as these were negative for all STS primers. When these results were combined with previous studies on 41 isolates from animals and compared with the diversity of genotypes of 102 human Blastocystis hominis isolates, 67.4 % (62/92) of isolates from mammals and birds were identical to human B. hominis genotypes. Since the unknown genotype of human origin had been placed into an additional clade in the small-subunit rRNA gene phylogeny, further molecular study on the eight isolates of unknown genotype from the present study will facilitate our understanding of their zoonotic potential. PMID- 15133075 TI - Phylogenetic identification of Pneumocystis murina sp. nov., a new species in laboratory mice. AB - Pneumocystis is a fungal genus that contains multiple species. One member of the genus that has not been formally analysed for its phylogenetic relationships and possible species status is the Pneumocystis found in laboratory mice, Pneumocystis murina sp. nov. (type strain ATCC PRA-111(T)=CBS 114898(T)), formerly known as Pneumocystis carinii f. sp. muris. To advance research in this area, approximately 3000 bp of additional DNA sequence were obtained from the locus encoding rRNAs. This sequence and others were used to determine genetic distances between P. murina and other members of the genus. These distances indicated that P. murina DNA is most similar to that of the species of Pneumocystis found in laboratory rats. Nevertheless, P. murina is at least as diverged from these other Pneumocystis species as species in other fungal genera are from each other. The 18S rRNA gene sequence divergence exhibited by P. murina could not be ascribed to accelerated evolution of this gene as similar levels of divergence were observed at seven other loci. When five genes were used to construct phylogenetic trees for five Pneumocystis taxa, including P. murina, all the trees had the same topology, indicating that genes do not flow among these taxa. The gene trees were all strongly supported by statistical tests. When sequences from the rRNA-encoding locus were used to estimate the time of divergence of P. murina, the results indicated that P. murina is as old as the mouse. Taken together, these data support previous recognition of multiple species in the genus and indicate that P. murina is a phylogenetic species as well. PMID- 15133076 TI - Molecular typing of Pneumocystis jirovecii found in formalin-fixed paraffin embedded lung tissue sections from sudden infant death victims. AB - Previous studies have provided histological evidence of an association between primary Pneumocystis infection and sudden infant death syndrome (SIDS). The aim of this work was to determine the species of clustered Pneumocystis organisms found in formalin-fixed paraffin-embedded (FFPE) lung tissue sections from Chilean sudden infant death (SID) victims. This approach needed first to optimize a DNA extraction method from such histological sections. For that purpose, the QIAamp DNA Isolation from Paraffin-Embedded Tissue method (Qiagen) was first tested on FFPE lung tissue sections of immunosuppressed Wistar rats inoculated with rat-derived PNEUMOCYSTIS: Successful DNA extraction was assessed by the amplification of a 346 bp fragment of the mitochondrial large subunit rRNA gene of the Pneumocystis species using a previously described PCR assay. PCR products were analysed by direct sequencing and sequences corresponding to Pneumocystis carinii were found in all the samples. This method was then applied to FFPE lung tissue sections from Chilean SID victims. Pneumocystis jirovecii was successfully identified in the three tested samples. In conclusion, an efficient protocol for isolating PCR-ready DNA from FFPE lung tissue sections was developed. It established that the Pneumocystis species found in the lungs of Chilean SID victims was P. jirovecii. PMID- 15133077 TI - Similar genotypes of Pneumocystis jirovecii in different forms of Pneumocystis infection. AB - This study describes the genotyping of Pneumocystis jirovecii organisms isolated from three groups of patients that developed diverse forms of P. jirovecii infection; the patients were monitored in the same French hospital. Forty archival specimens from 13 adults with Pneumocystis pneumonia, eight adults colonized by P. jirovecii and 19 immunocompetent infants infected with the fungus contemporaneously with a bronchiolitis episode were analysed retrospectively. Genotyping was performed by analysis of sequences of the internal transcribed spacer (ITS)1 and ITS2 regions, and of the dihydropteroate synthase (DHPS) locus. At the ITS regions, a high diversity of genotypes, identical main genotypes (B(1)a(3) and B(2)a(1)) and the occurrence of mixed infections (more than one genotype) were observed in the three patient groups. At the DHPS locus, the results indicated the presence of mutants in the two adult groups, as well as in the infant group. Consequently, at these two independent genomic regions, P. jirovecii isolates from patients who developed different forms of infection and who lived in the same geographical region presented common characteristics. These results suggest that patients infected with P. jirovecii, whatever the form of infection they present, are part of a common human reservoir for P. jirovecii. PMID- 15133078 TI - Inability of Pneumocystis organisms to incorporate bromodeoxyuridine suggests the absence of a salvage pathway for thymidine. AB - Because thymidine metabolism is a potential target for therapy of Pneumocystis pneumonia, it was investigated whether Pneumocystis organisms have a salvage pathway for thymidine by administering 5-bromo-2'-deoxyuridine (BrdU) to mice and rats with Pneumocystis pneumonia. Although BrdU incorporation was detected in host cells, no incorporation was seen in Pneumocystis organisms infecting either rats or mice. This suggests that Pneumocystis organisms do not have a salvage pathway for thymidine, and that inhibitors of de novo synthesis, such as thymidylate synthase inhibitors, may be effective drugs for treating Pneumocystis pneumonia. PMID- 15133079 TI - The plasma membrane of microaerophilic protists: oxidative and nitrosative stress. AB - The trans-plasma-membrane electrochemical potential of microaerophilic protists was monitored by the use of voltage-sensitive charged lipophilic fluorophores; of the many available probes, the anionic oxonol dye bis(1,3-dibarbituric acid) trimethine oxonol [DiBAC(4)(3)] is an example of one which has been successfully employed using fluorescence microscopy, confocal laser-scanning microscopy and flow cytometry. Several microaerophilic protists have been investigated with this dye; these were Giardia intestinalis, Trichomonas vaginalis, Tritrichomonas foetus, Hexamita inflata and Mastigamoeba punctachora. Under conditions where they exhibit normal vitality, these organisms exclude DiBAC(4)(3) by virtue of their maintenance of a plasma-membrane potential (negative inside). Uptake of the fluorophore is indicative of disturbance to this membrane (i.e. by inhibition of pump/leak balance, blockage of channels or generation of ionic leaks), and is indicative of metabolic perturbation or environmental stress. Here, it is shown that oxidative or nitrosative stress depolarizes the plasma membranes of the aforementioned O(2)-sensitive organisms and allows DiBAC(4)(3) influx. Oxonol uptake thereby provides a sensitive and early indication of plasma-membrane perturbation by agents that may lead to cytotoxicity and eventually to cell death by necrotic or apoptotic pathways. PMID- 15133080 TI - Intron-containing beta-tubulin transcripts in Cryptosporidium parvum cultured in vitro. AB - The genome of Cryptosporidium parvum contains a relatively small number of introns, which includes the beta-tubulin gene with only a single intron. Recently, it was observed that the intron was not removed from some of the beta tubulin transcripts in the late life cycle stages cultured in vitro. Although normally spliced beta-tubulin mRNA was detected in all parasite intracellular stages by RT-PCR (e.g. HCT-8 or Caco-2 cells infected with C. parvum for 12-72 h), at 48-72 h post-infection unprocessed beta-tubulin transcripts containing intact introns started to appear in parasite mRNA within infected host cells. The intron-containing transcripts could be detected by fluorescence in situ hybridization (FISH) using an intron-specific probe. The intron-containing beta tubulin transcripts appeared unique to the in vitro-cultured C. parvum, since they were not detected in parasite-infected calves at 72 h. As yet, it is unclear whether the late life cycle stages of C. parvum are partially deficient in intron splicing or the intron-splicing processes have merely slowed, both of which would allow the detection of intron-containing transcripts. Another possible explanation is that the decay in transcript processing might simply be due to the onset of parasite death. Nonetheless, the appearance of intron-containing transcripts coincides with the arrest of C. parvum development in vitro. This unusual observation prompts speculation that the abnormal intron-splicing of beta tubulin transcripts may be one of the factors preventing complete development of this parasite in vitro. Furthermore, the presence of both processed and unprocessed introns in beta-tubulin transcripts in vitro may provide a venue for studying overall mechanisms for intron-splicing in this parasite. PMID- 15133081 TI - Functional characterization of replication protein A2 (RPA2) from Cryptosporidium parvum. AB - Replication protein A (RPA) is a heterotrimeric complex of single-stranded DNA binding proteins that play multiple roles in eukaryotic DNA metabolism. The RPA complex is typically composed of heterologous proteins (termed RPA1, RPA2 and RPA3) in animals, plants and fungi, which possess different functions. Previously, two distinct, short-type RPA large subunits (CpRPA1 and CpRPA1B) from the apicomplexan parasite Cryptosporidium parvum were characterized. Here are reported the identification and characterization of a putative middle RPA subunit (CpRPA2) from this unicellular organism. Although the CpRPA2 gene encodes a predicted 40.1 kDa peptide, which is larger than other RPA2 subunits characterized to date, Western blot analysis of oocyst preparations detected a native CpRPA2 protein with a molecular mass of approximately 32 kDa, suggesting that CpRPA2 might undergo post-translational cleavage or the gene was translated at an alternative start codon. Immunofluorescence microscopy using a rabbit anti CpRPA2 antibody revealed that CpRPA2 protein was mainly distributed in the cytosol (rather than the nuclei) of C. parvum sporozoites. Semi-quantitative RT PCR data indicated that CpRPA2 was differentially expressed in a tissue culture model with highest expression in intracellular parasites infecting HCT-8 cells for 36 and 60 h. Sequence comparison suggests that RPA2 is a group of poorly conserved proteins. Nonetheless, functional analyses of recombinant proteins confirmed that CpRPA2 is a single-stranded DNA-binding protein and that it could serve as an in vitro phosphorylation target by a DNA-dependent protein kinase. The minimal length of poly(dT) required for CpRPA2 binding is 17 nucleotides, and the DNA-binding capability was inhibited by phosphorylation in vitro. These observations provide additional evidence on the divergence of RPA proteins between C. parvum and host, implying that the parasite DNA replication machinery could be explored as a chemotherapeutic target. PMID- 15133082 TI - Differential expression and interaction of transcription co-activator MBF1 with TATA-binding protein (TBP) in the apicomplexan Cryptosporidium parvum. AB - All gene-specific transcriptional activators initiate gene transcriptions by binding to promoter sequences and recruiting general transcription factors including TATA-binding protein (TBP) to upstream of targeted genes. Some of them require multiprotein bridging factors (MBFs); for example, the type 1 MBF (MBF1) which interconnects the gene activator with TBP. In this study, the properties of a previously cloned type 1 multiprotein bridging factor (CpMBF1) and a newly identified TBP (CpTBP1) from the apicomplexan Cryptosporidium parvum were investigated. Genes encoding both proteins were differentially expressed as determined by semi-quantitative RT-PCRs during the parasite life cycle, but in different patterns. The highest level of expression of CpMBF1 was in the well developed intracellular parasites, whereas that of CpTBP1 was found in intact oocysts and late intracellular stages, possibly correlated with the formation of oocysts. Both CpMBF1 and CpTBP1 were expressed as maltose-binding protein fusion proteins. The function of CpTBP1 was confirmed by its ability to bind a biotinylated DNA oligonucleotide containing TATA consensus sequence. The interaction between CpMBF1 and CpTBP1 was also observed by an electrophoretic mobility shift assay. Since little is known about the regulation and control of gene activity in C. parvum, this study may point to a new direction for the study of gene activation associated with the development of the complex life cycle of this parasite. PMID- 15133083 TI - Polyamine metabolism in a member of the phylum Microspora (Encephalitozoon cuniculi): effects of polyamine analogues. AB - The uptake, biosynthesis and catabolism of polyamines in the microsporidian parasite Encephalitozoon cuniculi are detailed with reference to the effects of oligoamine and arylamine analogues of polyamines. Enc. cuniculi, an intracellular parasite of mammalian cells, has both biosynthetic and catabolic enzymes of polyamine metabolism, as demonstrated in cell-free extracts of mature spores. The uptake of polyamines was measured in immature, pre-emergent spores isolated from host cells by Percoll gradient. Spermine was rapidly taken up and metabolized to spermidine and an unknown, possibly acetamidopropanal, by spermidine/spermine N(1)-acetyltransferase (SSAT) and polyamine oxidase (PAO). Most of the spermidine and the unknown product were found in the cell incubation medium, indicating they were released from the cell. bis(Ethyl) oligoamine analogues of polyamines, such as SL-11144 and SL-11158, as well as arylamine analogues [BW-1, a bis(phenylbenzyl) 3-7-3 analogue] blocked uptake and interconversion of spermine at micromolar levels and, in the case of BW-1, acted as substrate for PAO. The Enc. cuniculi PAO activity differed from that found in mammalian cells with respect to pH optimum, substrate specificity and sensitivity to known PAO inhibitors. SL-11158 inhibited SSAT activity with a mixed type of inhibition in which the analogue had a 70-fold higher affinity for the enzyme than the natural substrate, spermine. The interest in Enc. cuniculi polyamine metabolism and the biochemical effects of these polyamine analogues is warranted since they cure model infections of Enc. cuniculi in mice and are potential candidates for human clinical trials. PMID- 15133084 TI - Amino sugar phosphate levels in Giardia change during cyst wall formation. AB - The parasite Giardia intestinalis exists as a trophozoite (vegetative) that infects the human small intestine, and a cyst (infective) that is shed in host faeces. Cyst viability in the environment depends upon a protective cyst wall, which consists of proteins and a unique beta(1-3) GalNAc homopolymer. UDP-GalNAc, the precursor for this polysaccharide, is synthesized from glucose by an enzyme pathway that involves amino sugar phosphate intermediates. Using a novel method of microanalysis by capillary electrophoresis, the levels of amino sugar phosphate intermediates in trophozoites before encystment, during a period of active encystment and after the peak of encystment were measured. These levels were used to deduce metabolic control of amino sugar phosphates associated with encystment. Levels of amino sugar phosphate intermediates increased during encystment, and then decreased to nearly non-encysting levels. The most pronounced increase was in glucosamine 6-phosphate, which is the first substrate unique in this pathway, and which is the positive effector for the pathway's putative rate-controlling enzyme, UDP-GlcNAc pyrophosphorylase. Moreover, more UDP-GalNAc than UDP-GlcNAc, its direct precursor, was detected at 24 h. It is postulated that the enhanced UDP-GalNAc is a result of enhanced synthesis of UDP GlcNAc by the pyrophosphorylase, and its preferential conversion to UDP-GalNAc. These results suggest that kinetics of amino sugar phosphate synthesis in encysting Giardia favours the direction that supports cyst wall synthesis. The enzymes involved in synthesis of UDP-GalNAc and its conversion to cyst wall might be potential targets for therapeutic inhibitors of Giardia infection. PMID- 15133085 TI - Menadione kills trophozoites and cysts of Giardia intestinalis. AB - Production of reactive oxygen species by redox cycling in the presence of low levels of oxygen has been studied as a possible approach to anti-protozoal chemotherapeutic strategy. Incubation of the diplomonad flagellate Giardia intestinalis with 2-methy-1,4-naphthoquinone (menadione), under anaerobic conditions, gave UV absorption changes characteristic of reduction to menadiol; partial reversal was observed on admitting O(2). Under microaerobic conditions, similar to those on the surface of the jejunal mucosa, trophozoites consumed O(2) rapidly in the presence of menadione; reaction products included singlet O(2) (monitored by single photon counting of O(2)-dependent low-level chemiluminescence) and H(2)O(2) (measured by the formation of Complex I of microperoxidase). Trophozoites became swollen and incapable of regulatory volume control; these irreversible responses led to loss of motility, cessation of flagellar activity and cell death. Comparison of the sensitivities of trophozoites to metronidazole and menadione gave LC(50) values ( microg x ml(-1)) of 1.2 and 0.7, respectively; corresponding values for cysts (measured by in vitro excystation capacities) were >50 and 1.3. Menadione (LD(50) in mice, 0.5 g kg(-1)) is therefore a potentially more useful and general anti-giardial agent than metronidazole, as it is active against cysts as well as trophozoites. PMID- 15133086 TI - Cyst wall synthase: N-acetylgalactosaminyltransferase activity is induced to form the novel N-acetylgalactosamine polysaccharide in the Giardia cyst wall. AB - Uridine-5'-diphospho-N-acetylgalactosamine (UDP-GalNAc) is required in the formation of the outer filamentous wall of Giardia and is synthesized by inducible enzymes in the cytosol of encysting trophozoites. In this study, an inducible enzyme activity that is associated with a particle population isolated from encysting Giardia is reported, and this activity exclusively incorporates [1 (14)C]GalNAc (from UDP-[(14)C]GalNAc) into an ethanol precipitate with the same properties as the filamentous cyst wall of GIARDIA: This ethanol precipitate exhibits characteristics of Giardia cyst wall filaments in that both contain GalNAc as the only sugar moieties and are SDS-insoluble, proteinase- and alkali resistant and acid-hydrolysable. However, since the precise chemical nature of the ethanol precipitate remains unknown, this enzyme activity is referred to tentatively as cyst wall synthase (CWS). CWS activity peaks in cells between 24 and 36 h of encystment and exhibits a high affinity and marked specificity for UDP-GalNAc as its substrate. UDP-N-acetylglucosamine, UDP-glucose, UDP-galactose, D-glucosamine and D-galactosamine were not incorporated into the ethanol precipitate. Partially purified CWS activity exhibits an apparent K(m) of 0.048 mM for UDP-GalNAc, a V(max) of 0.70 nmol x min(-1) (mg protein)(-1) and a requirement for divalent cations in the following order of preference: Ca(2+), Mg(2+)>Co(2+)>>>Mn(2+), Zn(2+). EDTA inhibits CWS activity. PMID- 15133087 TI - Mitosomes of Entamoeba histolytica are abundant mitochondrion-related remnant organelles that lack a detectable organellar genome. AB - The existence of mitochondrion-related relict organelles (mitosomes) in the amitochondrial human pathogen Entamoeba histolytica and the detection of extranuclear DNA-containing cytoplasmic structures (EhKOs) has led to the suggestion that a remnant genome from the original mitochondrial endosymbiont might have been retained in this organism. This study reports on the mutually exclusive distribution of Cpn60 and extranuclear DNA in E. histolytica and on the distribution of Cpn60-containing mitosomes in this parasite. In situ nick translation coupled to immunofluorescence microscopy failed to detect the presence of DNA in mitosomes, either in fixed parasite trophozoites or in partially purified organellar fractions. These results indicate that a remnant organellar genome has not been retained in E. histolytica mitosomes and demonstrate unequivocally that EhKOs and mitosomes are distinct and unrelated cellular structures. PMID- 15133088 TI - Expression in fibroblasts and in live animals of Entamoeba histolytica polypeptides EhCP112 and EhADH112. AB - EhCPADH is an immunogenic, heterodimeric protein that is formed by EhCP112 (cysteine protease) and EhADH112 (adhesin), polypeptides involved in Entamoeba histolytica's cytopathic effect, target-cell adherence and phagocytosis. The EhCPADH complex is located in the plasma membrane and cytoplasmic vacuoles. Here, the independent expression of EhCP112 and EhADH112 in fibroblasts and hamsters was analysed. Also investigated was the immunological response in animals independently inoculated with plasmid pcDNA-Ehcp112, which carries the complete cysteine protease-encoding gene, or with plasmid pcDNA-Ehadh112, which carries the C terminus of the adhesin-encoding gene, or with a mixture of both. Both proteins were expressed in the plasma membranes of the transfected fibroblasts. EhCP112 was toxic for the mammalian cells. Proteins were also independently expressed in hamsters after inoculation with the plasmids. Their expression was indirectly evaluated by the presence of antibodies in the inoculated animals. Remarkably, co-immunization of the animals with the two DNA plasmids resulted in an earlier and higher anti-E. histolytica IgG induction than immunization with separate plasmids. In contrast, the cellular immune response was not noticeably improved by the plasmid mixture. Interestingly, protection against liver abscesses was detected only in animals that received the plasmid mixture and no protection was observed in hamsters independently inoculated with plasmid pcDNA Ehcp112 or pcDNA-Ehadh112. PMID- 15133089 TI - Spore morphotypes of Thelohania solenopsae (microsporidia) described microscopically and confirmed by PCR of individual spores microdissected from smears by position ablative laser microbeam microscopy. AB - Development of Thelohania solenopsae, a parasite of the red imported fire ant (Solenopsis invicta), until recently was thought to include formation of two types of spores: unicellular meiospores, maturing inside sporophorous vesicles in sets of eight (octospores); and Nosema-like binuclear free spores. Megaspores, discovered in 2001, develop primarily in alates and are morphologically distinct from the two previously known types of spores. The role of megaspores in the T. solenopsae life cycle, as well as their existence, has been questioned. The current research includes light and electron microscopic descriptions of the three major spore morphotypes characteristic of T. solenopsae development. In addition, individual octospores and megaspores were isolated into groups of 8-20 from methanol-fixed and Calcofluor-stained smears of the infected ants for subsequent PCR analysis by the laser pressure catapulting function of a position ablative laser microbeam microscope, a technique applied for the first time to research of microsporidia. The PCR-amplified SSU rDNA nucleotide sequences from octospores and megaspores were identical. This, along with the consistency with which megaspores are detected in infected ants, demonstrates that megaspores are integral to the life cycle of T. solenopsae. PMID- 15133090 TI - Effects of high light on transcripts of stress-associated genes for the cyanobacteria Synechocystis sp. PCC 6803 and Prochlorococcus MED4 and MIT9313. AB - Cyanobacteria constitute an ancient, diverse and ecologically important bacterial group. The responses of these organisms to light and nutrient conditions are finely controlled, enabling the cells to survive a range of environmental conditions. In particular, it is important to understand how cyanobacteria acclimate to the absorption of excess excitation energy and how stress-associated transcripts accumulate following transfer of cells from low- to high-intensity light. In this study, quantitative RT-PCR was used to monitor changes in levels of transcripts encoding chaperones and stress-associated proteases in three cyanobacterial strains that inhabit different ecological niches: the freshwater strain Synechocystis sp. PCC 6803, the marine high-light-adapted strain Prochlorococcus MED4 and the marine low-light-adapted strain Prochlorococcus MIT9313. Levels of transcripts encoding stress-associated proteins were very sensitive to changes in light intensity in all of these organisms, although there were significant differences in the degree and kinetics of transcript accumulation. A specific set of genes that seemed to be associated with high light adaptation (groEL/groES, dnaK2, dnaJ3, clpB1 and clpP1) could be targeted for more detailed studies in the future. Furthermore, the strongest responses were observed in Prochlorococcus MED4, a strain characteristic of the open ocean surface layer, where hsp genes could play a critical role in cell survival. PMID- 15133091 TI - Influences of temperature, salinity and starvation on the motility and chemotactic response of Vibrio anguillarum. AB - The role of growth factors for the motility and chemotaxis of the fish pathogen Vibrio anguillarum was determined. Cells of V. anguillarum were chemotactic to serine in the temperature range 5-25 degrees C and in 0.8-2.7 % NaCl. The chemotactic response was significantly higher at 25 degrees C than at 5 or 15 degrees C. Growth in medium with 1.5 % NaCl gave a higher response than growth with 3 % NaCl; when the salinity of the chemotaxis buffer was raised, the chemotactic response was reduced. The role of starvation was also studied; V. anguillarum showed a high chemotactic response after starvation for 2 and 8 days. Motility and chemotaxis are important virulence factors for this bacterium. Not only was the ability to perform chemotactic motility maintained after starvation, but also it was shown that starvation does not interfere with the ability of the organism to cause infection in rainbow trout after a bath challenge. The swimming speed was reduced at lower temperatures. Within the range of salinity and starvation studied, the motile cells swam with the same velocity, indicating that V. anguillarum under all the examined conditions has a functional flagellum and rotates it with constant speed. Phenamil, a specific inhibitor of Na(+)-driven flagella, reduced the motility of both starved and non-starved cells of V. anguillarum indicating that, in both cases, a Na(+) motive force drives the flagellum. PMID- 15133092 TI - Enhanced motility of a Proteus mirabilis strain expressing hybrid FlaAB flagella. AB - Proteus mirabilis has two tandemly arranged flagellin-encoding genes, flaA and flaB. flaA is transcribed from a sigma(28) promoter, while flaB is silent. flaA and flaB can undergo reversible rearrangement to produce a set of hybrid genes referred to as flaAB. Flagellins composed of FlaAB protein have a different amino acid sequence and are antigenically distinct from flagellin composed of FlaA, implicating flagellin gene conversion as a putative virulence mechanism for P. mirabilis. The change in amino acid sequence is also hypothesized to alter the filament helix and, hence, affect the motility of FlaAB-expressing strains. To test this hypothesis, the motility of wild-type P. mirabilis was compared with that of a strain, DF1003, locked into the FlaAB(+) hybrid phase, under conditions of altered ionic strength, pH and viscosity. Cell motion tracking analysis showed that DF1003 has wild-type swimming velocity at physiological conditions, but moves significantly faster and travels further compared to the wild-type at NaCl concentrations greater than 170 mM. DF1003 is also significantly faster than the wild-type at pH 5.2, 5.8 and 8.2, and at 5 and 10 % polyvinylpyrrolidone. Measurements of amplitude and wavelength for isolated flagella subjected to pH 5.8 or 425 mM NaCl showed a loss of helical structure in FlaA flagella compared to FlaAB filaments, a feature that could significantly affect motility under these conditions. These results support a hypothesis that FlaAB flagellin imparts a motile advantage to P. mirabilis in conditions that otherwise may impede bacterial movement. In a broader context, flagellar antigenic variation, commonly thought to serve as means to avoid host defences, may also enhance motility in other bacterial species, thus aiding in the adaptation and survival of the cells. PMID- 15133093 TI - NifH and NifD phylogenies: an evolutionary basis for understanding nitrogen fixation capabilities of methanotrophic bacteria. AB - The ability to utilize dinitrogen as a nitrogen source is an important phenotypic trait in most currently known methanotrophic bacteria (MB). This trait is especially important for acidophilic MB, which inhabit acidic oligotrophic environments, highly depleted in available nitrogen compounds. Phylogenetically, acidophilic MB are most closely related to heterotrophic dinitrogen-fixing bacteria of the genus BEIJERINCKIA: To further explore the phylogenetic linkage between these metabolically different organisms, the sequences of nifH and nifD gene fragments from acidophilic MB of the genera Methylocella and Methylocapsa, and from representatives of Beijerinckia, were determined. For reference, nifH and nifD sequences were also obtained from some type II MB of the alphaproteobacterial Methylosinus/Methylocystis group and from gammaproteobacterial type I MB. The trees constructed for the inferred amino acid sequences of nifH and nifD were highly congruent. The phylogenetic relationships among MB in the NifH and NifD trees also agreed well with the corresponding 16S rRNA-based phylogeny, except for two distinctive features. First, different methods used for phylogenetic analysis grouped the NifH and NifD sequences of strains of the gammaproteobacterial MB Methylococcus capsulatus within a clade mainly characterized by Alphaproteobacteria, including acidophilic MB and type II MB of the Methylosinus/Methylocystis group. From this and other genomic data from Methylococcus capsulatus Bath, it is proposed that an ancient event of lateral gene transfer was responsible for this aberrant branching. Second, the identity values of NifH and NifD sequences between Methylocapsa acidiphila B2 and representatives of Beijerinckia were clearly higher (98.5 and 96.6 %, respectively) than would be expected from their 16S rRNA-based relationships. Possibly, these two bacteria originated from a common acidophilic dinitrogen fixing ancestor, and were subject to similar evolutionary pressure with regard to nitrogen acquisition. This interpretation is corroborated by the observation that, in contrast to most other diazotrophs, M. acidiphila B2 and Beijerinckia spp. are capable of active growth on nitrogen-free media under fully aerobic conditions. PMID- 15133094 TI - Significant differences in type IV pilin allele distribution among Pseudomonas aeruginosa isolates from cystic fibrosis (CF) versus non-CF patients. AB - Type IV pili (TFP) are important colonization factors of the opportunistic pathogen Pseudomonas aeruginosa, involved in biofilm formation and attachment to host cells. This study undertook a comprehensive analysis of TFP alleles in more than 290 environmental, clinical, rectal and cystic fibrosis (CF) isolates of P. aeruginosa. Based on the results, a new system of nomenclature is proposed, in which P. aeruginosa TFP are divided into five distinct phylogenetic groups. Each pilin allele is stringently associated with characteristic, distinct accessory genes that allow the identification of the allele by specific PCR. The invariant association of the pilin and accessory genes implies horizontal transfer of the entire locus. Analysis of pilin allele distribution among isolates from various sources revealed a striking bias in the prevalence of isolates with group I pilin genes from CF compared with non-CF human sources (P<0.0001), suggesting this particular pilin type, which can be post-translationally modified by glycosylation via the action of TfpO (PilO), may confer a colonization or persistence advantage in the CF host. This allele was also predominant in paediatric CF isolates (29 of 43; 67.4 %), showing that this bias is apparent early in colonization. Group I pilins were also the most common type found in environmental isolates tested. To the authors' knowledge, this is the first example of a P. aeruginosa virulence factor allele that is strongly associated with CF isolates. PMID- 15133095 TI - Emergence of phenotypic variants upon mismatch repair disruption in Pseudomonas aeruginosa. AB - MutS is part of the bacterial mismatch repair system that corrects point mutations and small insertions/deletions that fail to be proof-read by DNA polymerase activity. In this work it is shown that the disruption of the P. aeruginosa mutS gene generates the emergence of diverse colony morphologies in contrast with its parental wild-type strain that displayed monomorphic colonies. Interestingly, two of the mutS morphotypes emerged at a high frequency and in a reproducible way and were selected for subsequent characterization. One of them displayed a nearly wild-type morphology while the other notably showed, compared with the wild-type strain, increased production of pyocyanin and pyoverdin, lower excretion of LasB protease and novel motility characteristics, mainly related to swarming. Furthermore, it was reproducibly observed that, after prolonged incubation in liquid culture, the pigmented variant consistently emerged from the mutS wild-type-like variant displaying a reproducible event. It is also shown that these P. aeruginosa mutS morphotypes not only displayed an increase in the frequency of antibiotic-resistant mutants, as described for clinical P. aeruginosa mutator isolates, but also generated mutants whose antibiotic resistant levels were higher than those measured from spontaneous resistant mutants derived from wild-type cells. It was also found that both morphotypes showed a decreased cytotoxic capacity compared to the wild-type strain, leading to the emergence of invasive variants. By using mutated versions of a tetracycline resistance gene, the mutS mutant showed a 70-fold increase in the reversion frequency of a +1 frameshift mutation with respect to its parental wild type strain, allowing the suggestion that the phenotypical diversity generated in the mutS population could be produced in part by frameshift mutations. Finally, since morphotypical diversification has also been described in clinical isolates, the possibility that this mutS diversification was related to the high frequency hypermutability observed in P. aeruginosa CF isolates is discussed. PMID- 15133096 TI - Stress-responsive proteins are upregulated in Streptococcus mutans during acid tolerance. AB - Streptococcus mutans is an important pathogen in the initiation of dental caries as the bacterium remains metabolically active when the environment becomes acidic. The mechanisms underlying this ability to survive and proliferate at low pH remain an area of intense investigation. Differential two-dimensional electrophoretic proteome analysis of S. mutans grown at steady state in continuous culture at pH 7.0 or pH 5.0 enabled the resolution of 199 cellular and extracellular protein spots with altered levels of expression. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry identified 167 of these protein spots. Sixty-one were associated with stress-responsive pathways involved in DNA replication, transcription, translation, protein folding and proteolysis. The 61 protein spots represented isoforms or cleavage products of 30 different proteins, of which 25 were either upregulated or uniquely expressed during acid-tolerant growth at pH 5.0. Among the unique and upregulated proteins were five that have not been previously identified as being associated with acid tolerance in S. mutans and/or which have not been studied in any detail in oral streptococci. These were the single-stranded DNA-binding protein, Ssb, the transcription elongation factor, GreA, the RNA exonuclease, polyribonucleotide nucleotidyltransferase (PnpA), and two proteinases, the ATP-binding subunit, ClpL, of the Clp family of proteinases and a proteinase encoded by the pep gene family with properties similar to the dipeptidase, PepD, of Lactobacillus helveticus. The identification of these and other differentially expressed proteins associated with an acid-tolerant-growth phenotype provides new information on targets for mutagenic studies that will allow the future assessment of their physiological significance in the survival and proliferation of S. mutans in low pH environments. PMID- 15133097 TI - Proteome analysis of Streptococcus mutans metabolic phenotype during acid tolerance. AB - Two-dimensional gel electrophoretic analysis of the proteome of Streptococcus mutans grown at a steady state in a glucose-limited anaerobic continuous culture revealed a number of proteins that were differentially expressed when the growth pH was lowered from pH 7.0 to pH 5.0. Changes in the expression of metabolic proteins were generally limited to three biochemical pathways: glycolysis, alternative acid production and branched-chain amino acid biosynthesis. The relative level of expression of protein spots representing all of the enzymes associated with the Embden-Meyerhof-Parnas pathway, and all but one of the enzymes involved in the major alternative acid fermentation pathways of S. mutans, was identified and measured. Proteome data, in conjunction with end product and cell-yield analyses, were consistent with a phenotypic change that allowed S. mutans to proliferate at low pH by expending energy to extrude excess H(+) from the cell, while minimizing the detrimental effects that result from the uncoupling of carbon flux from catabolism and the consequent imbalance in NADH and pyruvate production. The changes in enzyme levels were consistent with a reduction in the formation of the strongest acid, formic acid, which was a consequence of the diversion of pyruvate to both lactate and branched-chain amino acid production when S. mutans was cultivated in an acidic environment. PMID- 15133099 TI - Immunolocalization of NblA, a protein involved in phycobilisome turnover, during heterocyst differentiation in cyanobacteria. AB - In unicellular non-diazotrophic cyanobacteria, NblA is a small polypeptide required for phycobilisome degradation during macronutrient limitation. In the filamentous N(2)-fixing Tolypothrix sp., a nblA gene (nblAI) lies upstream of the cpeBA operon that encodes phycoerythrin apoproteins. Using a specific anti-NblAI antibody it was found that in strains of Tolypothrix sp. NblAI abundance increases under nitrogen-limiting conditions but the protein is also present in cells grown in nitrogen-replete medium. Gold immunolabelling experiments showed that, upon a nitrogen shift-down, NblAI is preferentially located in the differentiated heterocysts, where O(2) evolution has to be shut off for nitrogenase to operate. The results lead to the proposal that NblAI is a necessary 'cofactor' but not the triggering factor that governs phycobilisome degradation in Tolypothrix sp. PMID- 15133098 TI - Functional analysis of the transcriptional activator XlnR from Aspergillus niger. AB - The transcriptional activator XlnR from Aspergillus niger is a zinc binuclear cluster transcription factor that belongs to the GAL4 superfamily. Several putative structural domains in XlnR were predicted using database and protein sequence analysis. Thus far, only the functionality of the N-terminal DNA-binding domain has been determined experimentally. Deletion mutants of the xlnR gene were constructed to localize the functional regions of the protein. The results showed that a putative C-terminal coiled-coil region is involved in nuclear import of XlnR. After deletion of the C-terminus, including the coiled-coil region, XlnR was found in the cytoplasm, while deletion of the C-terminus downstream of the coiled-coil region resulted in nuclear import of XlnR. The latter mutant also showed increased xylanase activity, indicating the presence of a region with an inhibitory function in XlnR-controlled transcription. Previous findings had already shown that a mutation in the XlnR C-terminal region resulted in transcription of the structural genes under non-inducing conditions. A regulatory model of XlnR is presented in which the C-terminus responds to repressing signals, resulting in an inactive state of the protein. PMID- 15133101 TI - Strains of Bacillus cereus vary in the phenotypic adaptation of their membrane lipid composition in response to low water activity, reduced temperature and growth in rice starch. AB - The phenotypic adaptation of membrane lipids in seven strains of the food poisoning bacterium Bacillus cereus, isolated from Bangladeshi rice, is reported in relation to their ability to grow under conditions of low water activity (a(w)), reduced temperature and the presence of soluble rice starch. The strains have different membrane phospholipid head-group and fatty acyl compositions, and they display individual differences in their responses to both low a(w) and reduced temperature. The extent of the increase in anionic membrane lipids in response to low a(w) varies from strain to strain, is solute specific and in one strain does not occur. Growth is stimulated by the presence of soluble rice starch and results in a large rise in the proportion of diphosphatidylglycerol (DPG) at the expense of phosphatidylglycerol (PG), without any change in the proportion of total anionic phospholipids. Growth at 15 degrees C compared with 37 degrees C increases the proportions of DPG and phosphatidylethanolamine at the expense of PG. At the lower temperature there are changes in phospholipid fatty acyl composition characteristic of those expected to maintain membrane fluidity, including increases in the amount of total branched fatty acids and the anteiso /iso-branched ratio, and a decrease in the equivalent chain-length, but there are strain differences in how those changes were achieved. In contrast to some other bacilli, there are persistent large increases in the proportions of unsaturated fatty acyl chains in phospholipids during growth at 15 degrees C. PMID- 15133100 TI - The last step in coenzyme B(12) synthesis is localized to the cell membrane in bacteria and archaea. AB - In Salmonella enterica, the last step of the synthesis of adenosylcobamide is catalysed by the cobalamin synthase enzyme encoded by the cobS gene of this bacterium. Overexpression of the S. enterica cobS gene in Escherichia coli elicited the accumulation of the phage shock protein PspA, a protein whose expression has been linked to membrane stress. Resolution of inner and outer membranes of S. enterica by isopycnic density ultracentrifugation showed CobS activity associated with the inner membrane, a result that was confirmed using antibodies against CobS. Computer analysis of the predicted amino acid sequence of CobS suggested it was an integral membrane protein. Results of experiments performed with strains carrying plasmids encoding CobS-alkaline phosphatase or CobS-beta-galactosidase protein fusions were consistent with the membrane localization of the CobS protein. Modifications to the predicted model were made based on data obtained from experiments using protein fusions. The function encoded by the cobS orthologue in the methanogenic archaeon Methanobacterium thermoautotrophicum strain deltaH compensated for the lack of CobS during cobalamin synthesis in cobS strains of S. enterica. Cobalamin synthase activity was also detected in a membrane preparation of M. thermoautotrophicum. It was concluded that the assembly of the nucleotide loop of adenosylcobamides in archaea and bacteria is a membrane-associated process. Possible reasons for the association of adenosylcobamide biosynthetic enzymes with the cell membrane are discussed. PMID- 15133102 TI - Ammonium and hydroxylamine uptake and accumulation in Nitrosomonas. AB - Starved cells of Nitrosomonas europaea and further ammonia oxidizers were able to rapidly accumulate ammonium and hydroxylamine to an internal concentration of about 1 and 0.8 M, respectively. In kinetic studies, the uptake/accumulation rates for ammonium [3.1 mmol (g protein)(-1) min(-1)] and hydroxylamine [4.39 mmol (g protein)(-1) min(-1)] were determined. The uptake and accumulation process of ammonium and hydroxylamine was not coupled to ammonia or hydroxylamine oxidation and nitrite was not produced. In the presence of uncouplers the ammonium accumulation was completely inhibited, indicating an active, membrane potential-driven transport mechanism. When the external ammonium or hydroxylamine pool was depleted, the internal ammonium and hydroxylamine was consumed within 12 h or 20 min, respectively. The binding of ammonium/ammonia was correlated with an energized membrane system, and hydroxylamine may bind to the hydroxylamine oxidoredutase. PMID- 15133103 TI - Quantitative relationships for specific growth rates and macromolecular compositions of Mycobacterium tuberculosis, Streptomyces coelicolor A3(2) and Escherichia coli B/r: an integrative theoretical approach. AB - Further understanding of the physiological states of Mycobacterium tuberculosis and other mycobacteria was sought through comparisons with the genomic properties and macromolecular compositions of Streptomyces coelicolor A3(2), grown at 30 degrees C, and Escherichia coli B/r, grown at 37 degrees C. A frame of reference was established based on quantitative relationships observed between specific growth rates ( micro ) of cells and their macromolecular compositions. The concept of a schematic cell based on transcription/translation coupling, average genes and average proteins was developed to provide an instantaneous view of macromolecular synthesis carried out by cells growing at their maximum rate. It was inferred that the ultra-fast growth of E. coli results from its ability to increase the average number of rRNA (rrn) operons per cell through polyploidy, thereby increasing its capacity for ribosome synthesis. The maximum growth rate of E. coli was deduced to be limited by the rate of uptake and consumption of nutrients providing energy. Three characteristic properties of S. coelicolor A3(2) growing optimally ( micro =0.30 h(-1)) were identified. First, the rate of DNA replication was found to approach the rate reported for E. coli ( micro =1.73 h(-1)); secondly, all rrn operons were calculated to be fully engaged in precursor-rRNA synthesis; thirdly, compared with E. coli, protein synthesis was found to depend on higher concentrations of ribosomes and lower concentrations of aminoacyl-tRNA and EF-Tu. An equation was derived for E. coli B/r relating micro to the number of rrn operons per genome. Values of micro =0.69 h(-1) and micro =1.00 h(-1) were obtained respectively for cells with one or two rrn operons per genome. Using the author's equation relating the number of rrn operons per genome to maximum growth rate, it is expected that M. tuberculosis with one rrn operon should be capable of growing much faster than it actually does. Therefore, it is suggested that the high number of insertion sequences in this species attenuates growth rate to still lower values. PMID- 15133104 TI - The signal peptide sequence of a lytic transglycosylase of Neisseria meningitidis is involved in regulation of gene expression. AB - The 60 nucleotides encoding the signal peptide of the Neisseria meningitidis membrane-bound lytic transglycosylase (MltA) homologue GNA33 were found to exert a negative regulatory effect on expression of GNA33 from either a T7- or a P(lac) driven system in Escherichia coli. Down-regulation was observed to occur at the transcriptional/post-transcriptional level and could possibly be ascribed to the formation of a stem-loop secondary structure within the signal peptide sequence. Slowing down the transcription rate through inhibition/titration of the RNA polymerase resulted in a considerable increase in mRNA accumulation, suggesting that a better coupling of translation to transcription would impede the formation of the putative secondary structure. Screening of synonymous mutations in the signal peptide sequence that showed high-level expression of an in-frame fusion to a reporter resulted in the isolation of several deletion mutants lacking most of the sequence participating in the putative secondary structure. Interestingly, the increase in the steady-state mRNA level observed in deletion mutants was higher, reaching a 300-fold increment, than that found in substitution mutants. Our results support the hypothesis that the rate of transcription controls the formation of a secondary structure in the region of the GNA33 transcript corresponding to the signal peptide sequence and this, when formed, negatively regulates expression. PMID- 15133105 TI - Unique organization and regulation of the mrx fimbrial operon in Xenorhabdus nematophila. AB - Xenorhabdus nematophila, a Gram-negative bacterium belonging to the Proteus clade of the family Enterobacteriaceae, forms a mutualistic association with the soil nematode Steinernema carpocapsae. The nematode invades insects and releases Xenorhabdus into the haemolymph, where it participates in insect killing. To begin to understand the role of fimbriae in the unique life cycle of Xenorhabdus, the organization and expression of the mrx fimbrial operon was analysed. The mrx operon contained only five structural genes (mrxACDGH), making it one of the smallest chaperone-usher fimbrial operons studied to date. Unlike the mrp operon of Proteus mirabilis, a site-specific recombinase was not linked to the mrx operon. The intergenic region between the major fimbrial gene (mrxA) and the usher gene (mrxC) lacked a mrpB-like gene, but contained three tandem inverted repeat sequences located downstream of mrxA. A 940 nt mrxA-containing mRNA was the major transcript produced in cells growing on agar, while an mrx polycistronic mRNA was produced at low levels. A canonical sigma(70) promoter, identified upstream of mrxA, was not subject to promoter inversion. Fimbriae were not produced in an lrp-mutant strain, suggesting that the leucine-responsive regulatory protein, Lrp, plays a role in the regulation of the mrx operon. These findings show that the genetic organization and regulation of the mrx operon is in several respects distinct from other chaperone-usher fimbrial operons. PMID- 15133106 TI - The Fur-like protein Mur of Rhizobium leguminosarum is a Mn(2+)-responsive transcriptional regulator. AB - In wild-type Rhizobium leguminosarum, the sitABCD operon specifies a Mn(2+) transporter whose expression is severely reduced in cells grown in the presence of this metal. Mutations in the R. leguminosarum gene, mur (manganese uptake regulator), whose product resembles the Fur transcriptional regulator, cause high level expression of sitABCD in the presence of Mn(2+). In gel-shift mobility assays, purified R. leguminosarum Mur protein bound to at least two regions near the sitABCD promoter region, although this DNA has no conventional consensus Fur binding sequences (fur boxes). Thus, in contrast to gamma-proteobacteria, where Fur binds Fe(2+), the R. leguminosarum Fur homologue, Mur, act as a Mn(2) responsive transcriptional regulator. PMID- 15133107 TI - Increased transcription rates correlate with increased reversion rates in leuB and argH Escherichia coli auxotrophs. AB - Escherichia coli auxotrophs of leuB and argH were examined to determine if higher rates of transcription in derepressed genes were correlated with increased reversion rates. Rates of leuB and argH mRNA synthesis were determined using half lives and concentrations, during exponential growth and at several time points during 30 min of amino acid starvation. Changes in mRNA concentration were primarily due to increased mRNA synthesis and not to increased stability. Four strains of E. coli amino acid auxotrophs, isogenic except for relA and argR, were examined. In both the leuB and argH genes, rates of transcription and mutation were compared. In general, strains able to activate transcription with guanosine tetraphosphate (ppGpp) had higher rates of mRNA synthesis and mutation than those lacking ppGpp (relA2 mutants). argR knockout strains were constructed in relA(+) and relA mutant strains, and rates of both argH reversion and mRNA synthesis were significantly higher in the argR knockouts than in the regulated strains. A statistically significant linear correlation between increased rates of transcription and mutation was found for data from both genes. In general, changes in mRNA half-lives were less than threefold, whereas changes in rates of mRNA synthesis were often two orders of magnitude. The results suggest that specific starvation conditions target the biosynthetic genes for derepression and increased rates of transcription and mutation. PMID- 15133108 TI - Role of homoserine and threonine pathway intermediates as precursors for the biosynthesis of aminoethoxyvinylglycine in Streptomyces sp. NRRL 5331. AB - The genes hom, thrB and thrC, encoding homoserine dehydrogenase, homoserine kinase (HK) and threonine synthase, respectively, involved in the last steps of threonine biosynthesis, have been studied in Streptomyces sp. NRRL 5331, the producer of the ethylene synthetase inhibitor aminoethoxyvinylglycine (AVG), in order to determine their role in the biosynthesis of AVG. Different null mutants were obtained by plasmid-mediated disruption of each of the three genes. thrC gene disruption had no effect on AVG production, while the disruption of thrB blocked HK activity and substantially reduced the yield of this metabolite, probably due to the accumulation of homoserine and/or methionine which have a negative effect on AVG biosynthesis. Disruption of hom (thrA) completely blocked AVG biosynthesis, indicating that homoserine lies at the branching point of the aspartic-acid-derived biosynthetic route that leads to AVG. The four carbon atoms of the vinylglycine moiety of AVG derive, therefore, from homoserine. PMID- 15133109 TI - Molecular and functional characterization of type I signal peptidase from Legionella pneumophila. AB - Legionella pneumophila is a facultative intracellular Gram-negative rod-shaped bacterium that has become an important cause of both community-acquired and nosocomial pneumonia. Numerous studies concerning the unravelling of the virulence mechanism of this important pathogen have been initiated. As evidence is now accumulating for the involvement of protein secretion systems in bacterial virulence in general, the type I signal peptidase (LepB) of L. pneumophila was of particular interest. This endopeptidase plays an essential role in the processing of preproteins carrying a typical amino-terminal signal peptide, upon translocation across the cytoplasmic membrane. This paper reports the cloning and the transcriptional analysis of the L. pneumophila lepB gene encoding the type I signal peptidase (SPase). Reverse transcription PCR experiments showed clear lepB expression when L. pneumophila was grown both in culture medium, and also intracellularly in Acanthamoeba castellanii, a natural eukaryotic host of L. pneumophila. In addition, LepB was shown to be encoded by a polycistronic mRNA transcript together with two other proteins, i.e. a LepA homologue and a ribonuclease III homologue. SPase activity of the LepB protein was demonstrated by in vivo complementation analysis in a temperature-sensitive Escherichia coli lepB mutant. Protein sequence and predicted membrane topology were compared to those of leader peptidases of other Gram-negative human pathogens. Most strikingly, a strictly conserved methionine residue in the substrate binding pocket was replaced by a leucine residue, which might influence substrate recognition. Finally it was shown by in vivo experiments that L. pneumophila LepB is a target for (5S,6S)-6-[(R)-acetoxyethyl]-penem-3-carboxylate, a specific inhibitor of type I SPases. PMID- 15133110 TI - Two relA/spoT homologous genes are involved in the morphological and physiological differentiation of Streptomyces clavuligerus. AB - This study is focused on the involvement of the unusual nucleotide (p)ppGpp during the morphological and physiological differentiation of Streptomyces clavuligerus. In particular, the functional and structural elements of two genes encoding the proteins RelA and Rsh were identified. The relA gene encodes an 843 aa protein (RelA), while the rsh gene encodes a 738 aa protein (Rsh). The relA and rsh genes were disrupted by the insertion of a hygromycin resistance gene and an apramycin resistance gene, respectively. The synthesis of ppGpp in the relA gene-disrupted mutant was completely eliminated under conditions of starvation for amino acids, whereas synthesis persisted, but was greatly reduced in the rsh gene-disrupted mutant. The relA gene-disrupted mutant had a bald appearance on agar plate cultures and retarded growth in submerged culture, while the rsh disrupted mutant was unchanged in growth characteristics relative to the wild type culture. The production of both clavulanic acid and cephamycin C were completely abolished in the relA-disrupted mutant. Thus, it is concluded that the relA gene rather than rsh is essential for morphological and physiological differentiation in S. clavuligerus and that RelA primarily governs the stringent response of S. clavuligerus to starvation for amino acids. PMID- 15133111 TI - Properties of haemolysin E (HlyE) from a pathogenic Escherichia coli avian isolate and studies of HlyE export. AB - Haemolysin E (HlyE) is a novel pore-forming toxin first identified in Escherichia coli K-12. Analysis of the 3-D structure of HlyE led to the proposal that a unique hydrophobic beta-hairpin structure (the beta-tongue, residues 177-203) interacts with the lipid bilayer in target membranes. In seeming contradiction to this, the hlyE sequence from a pathogenic E. coli strain (JM4660) that lacks all other haemolysins has been reported to encode an Arg residue at position 188 that was difficult to reconcile with the proposed role of the beta-tongue. Here it is shown that the JM4660 hlyE sequence encodes Gly, not Arg, at position 188 and that substitution of Gly188 by Arg in E. coli K-12 HlyE abolishes activity, emphasizing the importance of the head domain in HlyE function. Nevertheless, 76 other amino acid substitutions were confirmed compared to the HlyE protein of E. coli K-12. The JM4660 HlyE protein was dimeric, suggesting a mechanism for improving toxin solubility, and it lysed red blood cells from many species by forming 36-41 A diameter pores. However, the haemolytic phenotype of JM4660 was found to be unstable due to defects in HlyE export, indicating that export of active HlyE is not an intrinsic property of the protein but requires additional components. TnphoA mutagenesis of hlyE shows that secretion from the cytoplasm to the periplasm does not require the carboxyl-terminal region of HlyE. Finally, disruption of genes associated with cell envelope function, including tatC, impairs HlyE export, indicating that outer membrane integrity is important for effective HlyE secretion. PMID- 15133112 TI - Induction of Mycobacterium avium growth restriction and inhibition of phagosome endosome interactions during macrophage activation and apoptosis induction by picolinic acid plus IFNgamma. AB - Treatment of mouse macrophages with picolinic acid (PA) and gamma-interferon (IFNgamma) led to the restriction of Mycobacterium avium proliferation concomitant with the sequential acquisition of metabolic changes typical of apoptosis, mitochondrial depolarization, annexin V staining and caspase activation, over a period of up to 5 days. However, triggering of cell death by ATP, staurosporine or H(2)O(2) failed to affect mycobacterial viability. In contrast to untreated macrophages where extensive interactions between phagosomes and endosomes were observed, phagosomes from treated macrophages lost the ability to acquire endosomal dextran. N-Acetylcysteine was able to revert both the anti mycobacterial activity of treated macrophages as well as the block in phagosome endosome interactions. The treatment, however, induced only a minor increase in the acquisition of lysosomal markers, namely Lamp-1, and did not increase to any great extent the acidification of the phagosomes. These data thus suggest that the anti-mycobacterial activity of PA and IFNgamma depends on the interruption of intracellular vesicular trafficking, namely the blocking of acquisition of endosomal material by the microbe. PMID- 15133114 TI - Molecular characterization of binding subcomponents of Clostridium botulinum type C progenitor toxin for intestinal epithelial cells and erythrocytes. AB - Clostridium botulinum type C 16S progenitor toxin consists of a neurotoxin (NTX), a non-toxic non-HA (NTNH), and a haemagglutinin (HA). The HA acts as an adhesin, allowing the 16S toxin to bind to intestinal epithelial cells and erythrocytes. In type C, these bindings are dependent on sialic acid. The HA consists of four distinct subcomponents designated HA1, HA2, HA3a and HA3b. To identify the binding subcomponent(s) of HA of type C 16S toxin, all of the HA-subcomponents and some of their precursor forms were produced as recombinant proteins fused to glutathione S-transferase (GST). These proteins were evaluated for their capacity to adhere to intestinal epithelial cells of guinea pig and human erythrocytes. GST-HA1, GST-HA3b and GST-HA3 (a precursor form of HA3a and HA3b) bound intestinal epithelial cells and erythrocytes, whereas GST alone, GST-HA2 and GST HA3a did not. GST-HA3b and GST-HA3 showed neuraminidase-sensitive binding to the intestinal epithelial cells and erythrocytes, whereas GST-HA1 showed neuraminidase-insensitive binding. TLC binding assay revealed that GST-HA3b and GST-HA3 recognized sialosylparagloboside (SPG) and GM3 in the ganglioside fraction of the erythrocytes, like native type C 16S toxin [Inoue, K. et al. (1999). Microbiology 145, 2533-2542]. On the other hand, GST-HA1 recognized paragloboside (PG; an asialo- derivative of SPG) in addition to SPG and GM3. Deletion mutant analyses of GST-HA3b showed that the C-terminal region of HA3b is important for its binding activity. Based on these data, it is concluded that the HA component contains two distinct carbohydrate-binding subcomponents, HA1 and HA3b, which recognize carbohydrates in different specificities. PMID- 15133113 TI - Genome structure in the vole bacillus, Mycobacterium microti, a member of the Mycobacterium tuberculosis complex with a low virulence for humans. AB - Mycobacterium microti, a member of the Mycobacterium tuberculosis complex, is phylogenetically closely related to M. tuberculosis, differing in a few biochemical properties. However, these species have different levels of virulence in different hosts; most notably M. microti shows lower virulence for humans than M. tuberculosis. This report presents genomic comparisons using DNA microarray analysis for an extensive study of the diversity of M. microti strains. Compared to M. tuberculosis H37Rv, 13 deletions were identified in 12 strains of M. microti, including the regions RD1 to RD10, which are also missing in Mycobacterium bovis BCG. In addition, four new deleted regions, named MiD1, RD1beta, MiD2 and MiD3, were identified. DNA sequencing was used to define the extent of most of the deletions in one strain. Although RD1 of M. bovis BCG and M. microti is thought to be crucial for attenuation, in this study, three of the four M. microti strains that were isolated from immunocompetent patients had the RD1 deletion. In fact, only the RD3 deletion was present in all of the strains examined, although deletions RD7, RD8 and MiD1 were found in almost all the M. microti strains. These deletions might therefore have some relation to the different host range of M. microti. It was also noticeable that of the 12 strains studied, only three were identical; these strains were all isolated from immunocompetent humans, suggesting that they could have arisen from a single source. Thus, this study shows that it is difficult to ascribe virulence to any particular pattern of deletion in M. microti. PMID- 15133115 TI - Evolution of multi-resistance plasmids in Australian clinical isolates of Escherichia coli. AB - Plasmids allow the movement of genetic material, including antimicrobial resistance genes, between bacterial species and genera. They frequently mediate resistance to multiple antimicrobials and can result in the acquisition by a pathogen of resistance to all or most clinically relevant antimicrobials. Unfortunately, there are still large gaps in our understanding of how new multi resistance plasmids evolve. Five Australian clinical institutions collaborated in this study of multi-resistance plasmids in clinical isolates of Escherichia coli. We characterized 72 resistance plasmids in terms of the antimicrobial resistance profile they conferred, their size and their incompatibility group. Restriction fragment length polymorphisms were used to determine the genetic relationships between the plasmids. Relationships between the host cells were determined using multi-locus enzyme electrophoresis. A lack of correlation between the evolutionary history of the host cells and their plasmids suggests that the horizontal transfer of resistance plasmids between strains of E. coli is common. The resistance plasmids were very diverse, with a wide range of resistance profiles and a lack of discrete evolutionary lineages. Multi-resistance plasmids did not evolve via the co-integrative capture of smaller resistance plasmids; rather, the roles of recombination and the horizontal movement of mobile genetic elements appeared to be most important. PMID- 15133116 TI - Cryptococcus neoformans Ilv2p confers resistance to sulfometuron methyl and is required for survival at 37 degrees C and in vivo. AB - Acetolactate synthase catalyses the first common step in isoleucine and valine biosynthesis and is the target of several classes of inhibitors. The Cryptococcus neoformans ILV2 gene, encoding acetolactate synthase, was identified by complementation of a Saccharomyces cerevisiae ilv2 mutant. C. neoformans is highly resistant to the commercially available acetolactate synthase inhibitor, sulfometuron methyl (SM). Expression of C. neoformans ILV2 in S. cerevisiae conferred SM resistance, indicating that the SM resistance of C. neoformans is due, at least in part, to C. neoformans Ilv2p. The C. neoformans ILV2 gene was disrupted. The ilv2 mutants were auxotrophic for isoleucine and valine and the auxotrophy was satisfied by these amino acids only when proline, and not ammonium, was the nitrogen source, indicating nitrogen regulation of amino acid transport. ilv2 mutants rapidly lost viability at 37 degrees C and when starved for isoleucine and valine. Consistent with these phenotypes, an ilv2 mutant was avirulent and unable to survive in mice. Because C. neoformans Ilv2p is required for virulence and survival in vivo, inhibitors of branched-chain amino acid biosynthesis may make valuable antifungal agents. PMID- 15133117 TI - SpeB modulates fibronectin-dependent internalization of Streptococcus pyogenes by efficient proteolysis of cell-wall-anchored protein F1. AB - SpeB is a cysteine proteinase and virulence determinant secreted by the important human pathogen Streptococcus pyogenes. Recent investigations have suggested a role for SpeB in streptococcal entry into human cells. However, conflicting data concerning the contribution of SpeB to internalization have been presented. Protein F1 is a cell-wall-attached fibronectin (Fn)-binding protein that is present in a majority of streptococcal isolates and is important for internalization. This study shows that protein F1 is efficiently degraded by SpeB, and that removal of protein F1 from the bacterial surface leads to reduced internalization. Whereas M1 protein and protein H, two additional surface proteins of S. pyogenes that bind human plasma proteins, are protected from proteolytic degradation by their ligands, protein F1 is readily cleaved by SpeB also when in complex with Fn. This finding, and the connection between the presence of Fn at the bacterial surface and entry into human cells, suggest that SpeB plays a role in the regulation of the internalization process. PMID- 15133118 TI - Linear versus circular mitochondrial genomes: intraspecies variability of mitochondrial genome architecture in Candida parapsilosis. AB - The yeast species Candida parapsilosis, an opportunistic pathogen, exhibits genetic and genomic heterogeneity. To assess the polymorphism at the level of mitochondrial DNA (mtDNA), the organization of the mitochondrial genome in strains belonging to the three variant groups of this species was investigated. Although these analyses revealed a group-specific restriction fragment pattern of mtDNA, strains belonging to different groups appear to have similar genes in the same gene order. An extensive survey of C. parapsilosis isolates uncovered surprising alterations in the molecular architecture of their mitochondrial genome. A screening strategy for strains harbouring mtDNA with rearranged architecture showed that nearly all strains from groups I and III possess linear mtDNA molecules terminating with arrays of tandem repeat units, while most of the group II strains have a circular mitochondrial genome. In addition, it was found that linear genophores in mitochondria of strains from different groups differ in the sequence of the mitochondrial telomeric repeat unit. The occurrence of altered forms of mtDNA among C. parapsilosis strains opens up the unique possibility to address questions concerning the evolutionary origin and replication strategy of linear and circular genomes in mitochondria. PMID- 15133119 TI - Global analysis of gene expression in an rpoN mutant of Listeria monocytogenes. AB - The role of the alternative sigma(54) factor, encoded by the rpoN gene, was investigated in Listeria monocytogenes by comparing the global gene expression of the wild-type EGDe strain and an rpoN mutant. Gene expression, using whole-genome macroarrays, and protein content, using two-dimensional gel electrophoresis, were analysed. Seventy-seven genes and nine proteins, whose expression was modulated in the rpoN mutant as compared to the wild-type strain, were identified. Most of the modifications were related to carbohydrate metabolism and in particular to pyruvate metabolism. However, under the conditions studied, only the mptACD operon was shown to be directly controlled by sigma(54). Therefore, the remaining modifications seem to be due to indirect effects. In parallel, an in silico analysis suggests that sigma(54) may directly control the expression of four different phosphotransferase system (PTS) operons, including mptACD. PTS activity is known to have a direct effect on the pyruvate pool and on catabolite regulation. These results suggest that sigma(54) is mainly involved in the control of carbohydrate metabolism in L. monocytogenes via direct regulation of PTS activity, alteration of the pyruvate pool and modulation of carbon catabolite regulation. PMID- 15133120 TI - Comparison of the structure-activity relationships of the integron-associated recombination sites attI3 and attI1 reveals common features. AB - Incorporation of gene cassettes into integrons occurs by IntI-mediated site specific recombination between a 59-base element (59-be) site in the cassette and an attI site in the integron. While the 59-be sites share common features and are recognized by several different IntI recombinases, the sequences of attI sites are not obviously related and are preferentially recognized by the cognate IntI. To determine the features of attI sites that are required for recombination proficiency, the structure-activity relationships of a second attI site, the attI3 site from the class 3 integron, were examined. The attI3 site was confined to within a region consisting of 68 bp from the integron backbone and 15 bp from the adjacent cassette. This region includes four IntI3-binding sites, as assessed by gel shift and methylation interference studies. Two of the binding sites are inversely oriented and constitute a simple site that includes the recombination crossover point. The two additional binding sites appear to be directly oriented and one of them is essential for efficient recombination of the attI3 site with a 59-be, but not for recombination with a second full-length attI3 site, which occurs at 100-fold lower frequency. The fourth site enhances attI3 with 59-be recombination 10-fold. The finding that the organization and overall properties of attI3 are very similar to those of attI1 indicates that these features are likely to be common to all attI sites. PMID- 15133122 TI - Crystal structures of a DNA octaplex with I-motif of G-quartets and its splitting into two quadruplexes suggest a folding mechanism of eight tandem repeats. AB - Recent genomic analyses revealed many kinds of tandem repeats of specific sequences. Some of them are related to genetic diseases, but their biological functions and structures are still unknown. Two X-ray structures of a short DNA fragment d(gcGA[G]1Agc) show that four base-intercalated duplexes are assembled to form an octaplex at a low K+ concentration, in which the eight G5 residues form a stacked double G-quartet in the central part. At a higher K+ concentration, however, the octaplex is split into just two halves. These structural features suggest a folding process of eight tandem repeats of d(ccGA[G]4Agg), according to a double Greek-key motif. Such a packaging of the repeats could facilitate slippage of a certain sequence during DNA replication, to induce increase or decrease of the repeats. PMID- 15133121 TI - The Saccharomyces cerevisiae gene CDC40/PRP17 controls cell cycle progression through splicing of the ANC1 gene. AB - The timing of events in the cell cycle is of crucial importance, as any error can lead to cell death or cancerous growth. This accurate timing is accomplished through the activation of specific CDC genes. Mutations in the CDC40/PRP17 gene cause cell cycle arrest at the G2/M stage. It was previously found that the CDC40 gene encodes a pre-mRNA splicing factor, which participates in the second step of the splicing reaction. In this paper we dissect the mechanism by which pre-mRNA splicing affects cell cycle progression. We identify ANC1 as the target of CDC40 regulation. Deletion of the ANC1 intron relieves the cell cycle arrest and temperature sensitivity of cdc40 mutants. Furthermore, we identify, through point mutation analysis, specific residues in the ANC1 intron that are important for its splicing dependency on Cdc40p. Our results demonstrate a novel mechanism of cell cycle regulation that relies on the differential splicing of a subset of introns by specific splicing factors. PMID- 15133123 TI - The cytokine receptor gp130 and its soluble form are under hormonal control in human endometrium and decidua. AB - The transmembrane protein gp130 plays a central role in cytokine action as a signal transducing receptor subunit common to all interleukin-6 type cytokines. Endometrial tissue obtained from women with a normal menstrual cycle and decidua obtained from women in the first or second trimester of pregnancy were assessed for gp130 by western blotting, immunohistochemistry and enzyme-linked immunosorbent assay (ELISA) analysis. By immunoblotting, two forms of gp130 were detected: one-the soluble form-of approximately 100 kDa and a larger membrane bound form of approximately 150 kDa. The latter became clearly visible in the mid to late secretory phase and was more pronounced in decidual tissue of second trimester compared to first trimester. Immunohistochemically, gp130 was located in glandular epithelial cells during the mid to late secretory phase, whereas staining in the proliferative phase was rather weak. In first and second trimester decidua, glandular cells were also positively stained. In addition, the invading trophoblast cells were gp130 positive. Soluble gp130 release was measured in the supernatants from primary endometrial and decidual cell cultures by ELISA and reached maximum values in cell cultures without addition of hormones. In cultured endometrial epithelial cells obtained during the proliferative phase of the cycle, the soluble gp130 release increased significantly under combined estradiol/progesterone supplementation which mimics the secretory phase conditions compared to estradiol supplementation alone. In cultured epithelial cells derived from decidual tissue of first trimester of pregnancy, similar effects of hormonal regulation were observed. Our results suggest that the balance between soluble gp130 and its membrane-bound form may play an important role in regulating cytokine action necessary for blastocyst implantation and for further interaction between the decidualized endometrium and the invading trophoblast. PMID- 15133124 TI - Simultaneous stretching and contraction of stress fibers in vivo. AB - To study the dynamics of stress fiber components in cultured fibroblasts, we expressed alpha-actinin and the myosin II regulatory myosin light chain (MLC) as fusion proteins with green fluorescent protein. Myosin activation was stimulated by treatment with calyculin A, a serine/threonine phosphatase inhibitor that elevates MLC phosphorylation, or with LPA, another agent that ultimately stimulates phosphorylation of MLC via a RhoA-mediated pathway. The resulting contraction caused stress fiber shortening and allowed observation of changes in the spacing of stress fiber components. We have observed that stress fibers, unlike muscle myofibrils, do not contract uniformly along their lengths. Although peripheral regions shortened, more central regions stretched. We detected higher levels of MLC and phosphorylated MLC in the peripheral region of stress fibers. Fluorescence recovery after photobleaching revealed more rapid exchange of myosin and alpha-actinin in the middle of stress fibers, compared with the periphery. Surprisingly, the widths of the myosin and alpha-actinin bands in stress fibers also varied in different regions. In the periphery, the banding patterns for both proteins were shorter, whereas in central regions, where stretching occurred, the bands were wider. PMID- 15133125 TI - Exclusion of lipid rafts and decreased mobility of CD94/NKG2A receptors at the inhibitory NK cell synapse. AB - CD94/NKG2A is an inhibitory receptor expressed by most human natural killer (NK) cells and a subset of T cells that recognizes human leukocyte antigen E (HLA-E) on potential target cells. To elucidate the cell surface dynamics of CD94/NKG2A receptors, we have expressed CD94/NKG2A-EGFP receptors in the rat basophilic leukemia (RBL) cell line. Photobleaching experiments revealed that CD94/NKG2A EGFP receptors move freely within the plasma membrane and accumulate at the site of contact with ligand. The enriched CD94/NKG2A-EGFP is markedly less mobile than the nonligated receptor. We observed that not only are lipid rafts not required for receptor polarization, they are excluded from the site of receptor contact with the ligand. Furthermore, the lipid raft patches normally observed at the sites where FcepsilonR1 activation receptors are cross-linked were not observed when CD94/NKG2A was coengaged along with the activation receptor. These results suggest that immobilization of the CD94/NKG2A receptors at ligation sites not only promote sustenance of the inhibitory signal, but by lipid rafts exclusion prevent formation of activation signaling complexes. PMID- 15133126 TI - Arabidopsis immunophilin-like TWD1 functionally interacts with vacuolar ABC transporters. AB - Previously, the immunophilin-like protein TWD1 from Arabidopsis has been demonstrated to interact with the ABC transporters AtPGP1 and its closest homologue, AtPGP19. Physiological and biochemical investigation of pgp1/pgp19 and of twd1 plants suggested a regulatory role of TWD1 on AtPGP1/AtPGP19 transport activities. To further understand the dramatic pleiotropic phenotype that is caused by loss-of-function mutation of the TWD1 gene, we were interested in other TWD1 interacting proteins. AtMRP1, a multidrug resistance-associated (MRP/ABCC) like ABC transporter, has been isolated in a yeast two-hybrid screen. We demonstrate molecular interaction between TWD1 and ABC transporters AtMRP1 and its closest homologue, AtMRP2. Unlike AtPGP1, AtMRP1 binds to the C-terminal tetratricopeptide repeat domain of TWD1, which is well known to mediate protein protein interactions. Domain mapping proved that TWD1 binds to a motif of AtMRP1 that resembles calmodulin-binding motifs; and calmodulin binding to the C terminus of MRP1 was verified. By membrane fractionation and GFP-tagging, we localized AtMRP1 to the central vacuolar membrane and the TWD1-AtMRP1 complex was verified in vivo by coimmunoprecipitation. We were able to demonstrate that TWD1 binds to isolated vacuoles and has a significant impact on the uptake of metolachlor-GS and estradiol-beta-glucuronide, well-known substrates of vacuolar transporters AtMRP1 and AtMRP2. PMID- 15133127 TI - IRI-1, a LIN-15B homologue, interacts with inositol-1,4,5-triphosphate receptors and regulates gonadogenesis, defecation, and pharyngeal pumping in Caenorhabditis elegans. AB - Inositol-1,4,5-triphosphate receptors (IP(3)Rs) are ligand-gated Ca(2+) channels that control Ca(2+) release from intracellular stores. They are central to a wide range of cellular responses. IP(3)Rs in Caenorhabditis elegans are encoded by a single gene, itr-1, and are widely expressed. Signaling through IP(3) and IP(3)Rs is important in ovulation, control of the defecation cycle, modulation of pharyngeal pumping rate, and embryogenesis. To further elucidate the molecular basis of the diversity of IP(3)R function, we used a yeast two-hybrid screen to search for proteins that interact with ITR-1. We identified an interaction between ITR-1 and IRI-1, a previously uncharacterized protein with homology to LIN-15B. Iri-1 is widely expressed, and its expression overlaps significantly with that of itr-1. In agreement with this observation, iri-1 functions in known itr-1-mediated processes, namely, upregulation of pharyngeal pumping in response to food and control of the defecation cycle. Knockdown of iri-1 in an itr-1 loss of-function mutant potentiates some of these effects and sheds light on the signaling pathways that control pharyngeal pumping rate. Knockdown of iri-1 expression also results in a sterile, evl phenotype, as a consequence of failures in early Z1/Z4 lineage divisions, such that gonadogenesis is severely disrupted. PMID- 15133128 TI - Use of fluorescent protein tags to study nuclear organization of the spliceosomal machinery in transiently transformed living plant cells. AB - Although early studies suggested that little compartmentalization exists within the nucleus, more recent studies on metazoan systems have identified a still increasing number of specific subnuclear compartments. Some of these compartments are dynamic structures; indeed, protein and RNA-protein components can cycle between different domains. This is particularly evident for RNA processing components. In plants, lack of tools has hampered studies on nuclear compartmentalization and dynamics of RNA processing components. Here, we show that transient expression of fluorescent protein fusions of U1 and U2 small nuclear ribonucleoprotein particle (snRNP)-specific proteins U1-70K, U2B", and U2A ', nucleolar proteins Nop10 and PRH75, and serine-arginine-rich proteins in plant protoplasts results in their correct localization. Furthermore, snRNP specific proteins also were correctly assembled into mature snRNPs. This system allowed a systematic analysis of the cellular localization of Arabidopsis serine arginine-rich proteins, which, like their animal counterparts, localize to speckles but not to nucleoli and Cajal bodies. Finally, markers for three different nuclear compartments, namely, nucleoli, Cajal bodies, and speckles, have been established and were shown to be applicable for colocalization studies in living plant protoplasts. Thus, transient expression of proteins tagged with four different fluorescent proteins is a suitable system for studying the nuclear organization of spliceosomal proteins in living plant cells and should therefore allow studies of their dynamics as well. PMID- 15133129 TI - SGEF, a RhoG guanine nucleotide exchange factor that stimulates macropinocytosis. AB - SGEF (SH3-containing Guanine Nucleotide Exchange Factor) is a RhoGEF of unknown function. We found the SGEF protein to be expressed in many established cell lines and highly expressed in human liver tissue. SGEF stimulated the formation of large interconnected membrane ruffles across dorsal surfaces when expressed in fibroblasts. SGEF required its proline-rich amino-terminus to generate dorsal, but not lateral, membrane ruffles and a functional SH3 domain to colocalize with filamentous actin at sites of membrane protrusion. Full-length SGEF activated RhoG, but not Rac, when expressed in fibroblasts. Further, recombinant SGEF DH/PH protein exchanged nucleotide on RhoG, but not on Rac1 or Rac3, in vitro. Scanning electron microscopy of fibroblasts demonstrated that SGEF induced dorsal ruffles that were morphologically similar to those generated by constitutively active RhoG, but not constitutively active Rac1. Transient expression of SGEF stimulated fibroblast uptake of 10-kDa dextran, a marker of macropinocytosis. This required the full-length protein and a catalytically active DH domain. Finally, activated RhoG was found to be more effective than activated Rac, and comparable to SGEF, in its ability to trigger dextran uptake. Together, this work establishes SGEF as a RhoG exchange factor and provides evidence that both SGEF and RhoG regulate membrane dynamics in promotion of macropinocytosis. PMID- 15133130 TI - Peroxisomal membrane proteins contain common Pex19p-binding sites that are an integral part of their targeting signals. AB - Targeting of peroxisomal membrane proteins (PMPs) is a multistep process that requires not only recognition of PMPs in the cytosol but also their insertion into the peroxisomal membrane. As a consequence, targeting signals of PMPs (mPTS) are rather complex. A candidate protein for the PMP recognition event is Pex19p, which interacts with most PMPs. However, the respective Pex19p-binding sites are ill-defined and it is currently disputed whether these sites are contained within mPTS. By using synthetic peptide scans and yeast two-hybrid analyses, we determined and characterized Pex19p-binding sites in Pex11p and Pex13p, two PMPs from Saccharomyces cerevisiae. The sites turned out to be composed of a short helical motif with a minimal length of 11 amino acids. With the acquired data, it proved possible to predict and experimentally verify Pex19p-binding sites in several other PMPs by applying a pattern search and a prediction matrix. A peroxisomally targeted Pex13p fragment became mislocalized to the endoplasmic reticulum in the absence of its Pex19p-binding site. By adding the heterologous binding site of Pex11p, peroxisomal targeting of the Pex13p fragment was restored. We conclude that Pex19p-binding sites are well-defined entities that represent an essential part of the mPTS. PMID- 15133131 TI - Na,K-ATPase beta1-subunit increases the translation efficiency of the alpha1 subunit in MSV-MDCK cells. AB - The Na,K-ATPase consists of an alpha- and beta-subunit. Moloney sarcoma virus transformed MDCK cells (MSV-MDCK) express low levels of Na,K-ATPase beta(1) subunit. Ectopic expression of Na,K-ATPase beta(1)-subunit in these cells increased the protein levels of the alpha(1)-subunit of Na,K-ATPase. This increase was not due to altered transcription of the alpha(1)-subunit gene or half-life of the alpha(1)-subunit protein because both alpha(1)-subunit mRNA levels and half-life of the alpha(1)-subunit protein were comparable in MSV-MDCK and beta(1)-subunit expressing MSV-MDCK cells. However, short pulse labeling revealed that the initial translation rate of the alpha(1)-subunit in beta(1) subunit expressing MSV-MDCK cells was six- to sevenfold higher compared with MSV MDCK cells. The increased translation was specific to alpha(1)-subunit because translation rates of occludin and beta-catenin, membrane and cytosolic proteins, respectively, were not altered. In vitro cotranslation/translocation experiments using rabbit reticulocyte lysate and rough microsomes revealed that the alpha(1) subunit mRNA is more efficiently translated in the presence of beta(1)-subunit. Furthermore, sucrose density gradient analysis revealed significantly more alpha(1)-subunit transcript associated with the polysomal fraction in beta(1) subunit expressing MSV-MDCK cells compared with MSV-MDCK cells, indicating that in mammalian cells the Na,K-ATPase beta(1)-subunit is involved in facilitating the translation of the alpha(1)-subunit mRNA in the endoplasmic reticulum. PMID- 15133133 TI - Editor's choice: evidence-not event-based practice. PMID- 15133132 TI - Clathrin isoform CHC22, a component of neuromuscular and myotendinous junctions, binds sorting nexin 5 and has increased expression during myogenesis and muscle regeneration. AB - The muscle isoform of clathrin heavy chain, CHC22, has 85% sequence identity to the ubiquitously expressed CHC17, yet its expression pattern and function appear to be distinct from those of well-characterized clathrin-coated vesicles. In mature muscle CHC22 is preferentially concentrated at neuromuscular and myotendinous junctions, suggesting a role at sarcolemmal contacts with extracellular matrix. During myoblast differentiation, CHC22 expression is increased, initially localized with desmin and nestin and then preferentially segregated to the poles of fused myoblasts. CHC22 expression is also increased in regenerating muscle fibers with the same time course as embryonic myosin, indicating a role in muscle repair. CHC22 binds to sorting nexin 5 through a coiled-coil domain present in both partners, which is absent in CHC17 and coincides with the region on CHC17 that binds the regulatory light-chain subunit. These differential binding data suggest a mechanism for the distinct functions of CHC22 relative to CHC17 in membrane traffic during muscle development, repair, and at neuromuscular and myotendinous junctions. PMID- 15133134 TI - Psychiatric disorder and the role of occupational health. PMID- 15133135 TI - Occupational medicine: the use of physician assistants and the changing role of the occupational and environmental medicine provider. PMID- 15133136 TI - Occupational health services in the UK--challenges and opportunities. AB - BACKGROUND: A small minority of the UK workforce currently has access to an occupational physician. Reduction in the size of enterprises, the emergence of atypical work patterns and problems recruiting and training occupational health specialists risk making this minority even smaller. AIM: This paper considers the challenges currently facing occupational medicine and how we can improve access to occupational health services (OHS). It aims to highlight some of the diverse internal and external factors that restrict the UK's ability to provide all workers access to OHS. METHOD: A literature review was carried out and combined with awareness of current trends in business and new legislation together with provision of occupational medicine in other countries. RESULTS: Potentially controversial solutions that might help to make OHS more widely accessible were identified and are discussed. It is hoped that these will provoke further debate. CONCLUSION: Individually and organizationally, we must examine and improve capabilities if we are to improve worker access to OHS and deliver targets to reduce occupational ill-health. It is suggested that this requires a strategic shift to apply resources differently. There is need to explore delegation of tasks traditionally performed by doctors to nurses and other staff together with the outsourcing of non-core work. The increased use of telemedicine and the enhanced use of information technology for training, risk assessments, wellness programmes and questionnaire-based health assessments are other developments that should be explored. PMID- 15133137 TI - Physician assistants in occupational medicine: how do they compare to occupational physicians? AB - BACKGROUND: Physician assistants (PAs) have been present in occupational and environmental medicine (OEM) in the USA since 1971, yet remarkably little is known about their activity. METHODS: An administrative study of PA activities was undertaken and compared with the activities of physicians in the same occupational medicine setting. Patients were not triaged to either provider and all resources of care were recorded for the visit. An episode of care approach was used for the analysis. RESULTS: The characteristics of patients seen by each provider were similar in age, gender ratio and severity of injury. Physicians saw a mean of 2.9 patients/h and PAs 2.5, but PAs worked more hours and saw more patients per year than physicians. The average charge per patient visit and total charge for an episode of care were similar. Differences between PAs and physicians were seen in the areas of 'limited duty' duration given to patients and on average PAs prescribed 15 days and physicians 17 days. PAs referred a patient 19.7% of the time, while physicians referred 17.4%. Most of the referrals were to physical therapy. The salary of a physician, based on an hourly rate, was approximately twice as much as a PA. CONCLUSION: The use of PAs in OEM may represent a cost-effective advantage from an administrative standpoint. Clearly, more research is necessary in determining the role and utilization of PAs in OEM and how they may improve the delivery of physician services. PMID- 15133138 TI - An evaluation of occupational health services within the NHS in London. AB - AIMS: To establish the nature, extent and organization of occupational health service provision for employees within the National Health Service (NHS) in London and to review the systems for monitoring performance. METHODS: Human resources directors and occupational health managers were contacted from a random selection of NHS trusts in the London area and invited to complete an interviewer led questionnaire. RESULTS: All seventeen trusts interviewed claimed to provide an occupational health service to their employees, with 88% providing this service in-house. The organization of the services varied, although most resided within the human resources function. Only 29% of the trusts could provide a written occupational health policy. Teaching hospital trusts had the most qualified and the highest numbers of medical staff. District/General hospital trusts had the least qualified clinical staff. Although most trusts were able to provide a comprehensive range of services, 87% of occupational health managers felt they could only provide a reactive service. Income was generated from non NHS sources by 88% of the trusts and all were aware of NHS Plus. There was an indication that some trusts assigned NHS Plus status did not meet the standard of NHS Plus, although the survey took place only 3 months after the launch of NHS Plus. CONCLUSIONS: There was a significant variation in the nature and extent of occupational health services in the NHS trusts. As a consequence, there may be differences in the level of occupational health service available to staff across the NHS in London. PMID- 15133139 TI - Charging for NHSPlus: an inferential study based on the internal provision of occupational health services within the National Health Service. AB - BACKGROUND: 'NHSPlus' was conceived as a national agency that would provide occupational health services to organizations, for a fee, without imposing any financial burden on the taxpayer. This self-funding requirement brings into focus the resource implications for such a service and the determination of the charges to be made to external clients. AIM: The existing provision of occupational health services to >100000 National Health Service (NHS) staff by 13 NHS occupational health services of various sizes was analysed, with the objective of determining an appropriate charge-out rate to third parties. METHOD: Two focus groups were questioned on their work external to the NHS. Data collected on the allocation of doctors and nurses to occupational health services in relation to the number of NHS clients serviced were used to investigate the nature of the resourcing relationship using regression analysis. RESULTS: The relationship was found to be stable enough to provide a good estimate of staff requirements (the key resource requirement). Combining this with costing information allowed inferences to be drawn concerning the economic cost and hence the break-even rate of charge for the service. This was then compared with the employer charge rates in the NHSPlus published case studies. CONCLUSIONS: The results suggest that the per capita charges to external clients are lower than the per capita cost of internal occupational health provision within the NHS, raising questions about the viability of the service. PMID- 15133140 TI - What do consultant occupational physicians do in the National Health Service? An audit. AB - AIM: To assess the range of activities undertaken by National Health Service (NHS) consultant occupational physicians, and quantify the proportion of time spent on these so that appropriate guidance and a model job description may be developed by the Association of NHS Occupational Physicians (ANHOPS). METHODS: A questionnaire was developed and sent to all full time consultant occupational health physicians (as recorded on the ANHOPS database). RESULTS: Sixty-five questionnaires were sent out, of which 38 were returned (59%). Only 31(48%) of replies matched the entry criteria of being full-time NHS consultants returning fully completed questionnaires. An average of 18 h face-to-face clinical work (range = 0-36 h) over five sessions (range = 0-9 sessions) was found. Remaining time was predominantly spent on clinical administrative work although some consultants took on a variety of other managerial activities. CONCLUSIONS: Responding full-time NHS consultant OHPs found it difficult to quantify and categorize their workloads, particularly for non-clinical work. Their responsibilities vary widely. Of use for job planning purposes is the average commitment of clinical sessions (five). PMID- 15133141 TI - Is there value in routinely obtaining a report from the general practitioner as part of pre-entry health screening of students for nursing studies? AB - BACKGROUND: Reports from general practitioners (GPs) are requested on applicants for nurse training, but there is no published evidence of the merit of this practice. AIMS: To assess the benefit of GP report in health assessments of student nurse applicants. METHODS: An audit was made of information obtained by health declaration form (HDF), nurse's assessment, GP report and, when performed, a physician's assessment for each applicant. Agreement between the health questionnaire and GP report was analysed by kappa statistics. RESULTS: Of 254 applicants, 246 (97%) were declared 'fit to work', four (1.6%) were deemed 'fit with restrictions' and four (1.6%) were considered 'unfit to work'. The most common problems declared were psychiatric and skin problems. The agreement between health declaration and the information provided by GPs was classed as almost perfect for diabetes and only fair to moderate for all other measures. The reports provided additional information on problems not declared by applicants, but all of these were passive problems. The four unfit candidates all had psychiatric illness, but in all cases the occupational health assessment was sufficient to make this decision or to request further information. In the 'fit with restrictions' category, three of the four GP reports (75%) helped in correctly assigning the applicants to this category. In one of these eight cases a passive problem had not been declared. CONCLUSIONS: The additional information in GP reports does not affect the conclusion regarding fitness for training in most cases and does not provide sufficient information to merit it being sought routinely. PMID- 15133142 TI - Physicians as clinical directors: working conditions, psychosocial resources and self-rated health. AB - BACKGROUND: Physicians in clinical directors' positions fulfil their commitments in demanding work environments characterized by organizational changes and economic cutbacks. Little is known about the self-rated health of this group. AIM: To investigate whether self-rated health was associated with psychosocial working conditions, professional networks, job support, social networks and social support, sick leave and salary in Swedish physicians working as clinical directors. METHODS: A self-reported questionnaire was sent to 373 clinical directors. Odds ratios (ORs) were used for estimating the bivariate association between self-rated health and psychosocial resources. RESULTS: A total of 274 clinical directors agreed to participate in the study. The response rate was 73%. The clinical directors exposed to high job demands had a significantly higher probability of low self-rated health [OR = 3.4 and 95% confidence interval (CI) = 1.6-7.0] than those who were not in this situation. Furthermore, participants who were exposed to high job demands had an increased risk of low self-rated health (OR = 3.8 and 95% CI = 1.8-8.1) irrespective of available social support inside or outside work. High average working hours more than doubled the risk of low self-rated health (OR = 2.2 and 95% CI = 1.1-4.4). CONCLUSION: The job demands on physicians in clinical directors' positions may exceed ordinary means of support with consequent adverse effects on self-rated health. More research is needed to investigate the interaction between job demands and support systems in this group of health care workers. PMID- 15133143 TI - Psychological impact of the 2003 severe acute respiratory syndrome outbreak on health care workers in a medium size regional general hospital in Singapore. AB - AIMS: To describe the psychological impact of severe acute respiratory syndrome (SARS) on health care workers in a regional general hospital 2 months post outbreak. METHOD: Doctors and nurses were encouraged to participate. The survey consisted of self-report measures: demographics, the General Health Questionnaire (GHQ) 28 and Impact of Events Scale (IES). A questionnaire enquiring about changes in life's priorities due to SARS and circumstances that helped with coping was used. Participation was strictly voluntary and responses anonymous. RESULTS: In total 177 out of 661 (27%) participants [40 out of 113 (35%) doctors and 137 out of 544 (25%) nurses] had a GHQ 28 score >or=5. Doctors [P = 0.026, odds ratio (OR) = 1.6 and 95% confidence interval (CI) = 1.1-2.5] and single health care workers were at higher risk (P = 0.048, OR = 1.4 and 95% CI = 1.02 2.0) compared to nurses and those who were married. Approximately 20% of the participants had IES scores >or=30, indicating the presence of post-traumatic stress disorder (PTSD). Four areas were classified as more important using factor analysis: health and relationship with the family, relationship with friends/colleagues, work and spiritual. The areas for coping strategies were clear directives/precautionary measures, ability to give feedback to/obtain support from management, support from supervisors/colleagues, support from the family, ability to talk to someone and religious convictions. Support from supervisors/colleagues was a significant negative predictor for psychiatric symptoms and PTSD. Work and clear communication of directives/precautionary measures also helped reduce psychiatric symptoms. CONCLUSIONS: Many health care workers were emotionally affected and traumatized during the SARS outbreak. Hence, it is important for health care institutions to provide psychosocial support and intervention for their health care workers. PMID- 15133144 TI - Factors influencing uptake of influenza vaccination among hospital-based health care workers. AB - BACKGROUND: Vaccination of health care workers against influenza has been shown to lower mortality among elderly patients, but uptake of voluntary vaccination among health care workers remains low. AIMS: Factors influencing uptake of vaccination were examined among a cross-section of health care workers based in an NHS Trust. METHODS: A structured, self-administered questionnaire was mailed to a random sample of health care workers based in the acute services sector of a UK National Health Service Trust, 6 months following a voluntary immunization programme implemented as part of the Scottish Executive Health Department winter planning arrangements for 2000-2001. The programme was promoted using posters in clinical areas and a single leaflet given to all staff through a paycheck advice note. RESULTS: Five hundred and fifty-one health care workers (53%) responded to the questionnaire and influenza vaccination was accepted by 150 (28%). The occupational health poster strongly influenced the decision to accept vaccination [odds ratio (OR) = 11.01; 95% confidence interval (CI) = 2.13-56.80; P < 0.0001]. Other significant influences included female sex (OR = 9.11; 95% CI = 1.26-65.72) and perceived risk of contracting flu without the vaccine (OR = 7.70; 95% CI = 1.44-41.05). Misconceptions regarding the purpose of the vaccination campaign were common and concern regarding possible side-effects was a deterring factor for vaccination uptake. CONCLUSION: Our study showed that visual material displayed throughout the workplace strongly influenced the acceptance of influenza vaccination. Future campaigns should also emphasize the positive benefits to patients of health care worker immunization, with readily accessible information regarding side-effects available from all sources. PMID- 15133145 TI - Unrecognized abrasions and occupational exposures to blood-borne pathogens among health care workers in Turkey. AB - AIMS: To evaluate the prevalence of exposures to potentially infective biological material, the use of gloves and the presence and awareness of skin abrasions amongst health care workers (HCWs). METHODS: Analysis of reported exposures during a 12 month period, questionnaire and skin examination by a dermatologist on HCWs at the University Hospital in Duzce, Turkey. RESULTS: Out of 415 HCWs, 278 responded to the questionnaire giving a response rate of 67%. There were 152 potentially infectious exposures reported on the questionnaire. Only 14% (21) of exposures were recorded between October 2001 and October 2002 at the time of the incident. Sharps injuries (57%) were reported most frequently in nurses, while splashes to mucous membranes occurred most frequently in physicians (36%). The operating theatre was the major location of incidents (56%). The frequency of abrasions on the hands was higher in female HCWs (60%) (P < 0.05). Wearing gloves for all procedures was most common in laboratory workers (60%). Abrasions were found on the hands of 142 (51%) of 278 HCWs. Sixteen per cent of the HCWs were unaware of abrasions on their hands. CONCLUSION: There appears to be under reporting of potentially infectious exposures by HCWs. HCWs are not always aware of abrasions on their hands. This study reinforces the need to report exposures and to use personal protective equipment. PMID- 15133146 TI - Influence of work environment on emotional health in a health care setting. AB - BACKGROUND: Radical changes are taking place in health care services and might be expected to cause job dissatisfaction, absenteeism, somatic complaints and mental health problems. Research in this area is limited and focused primarily on nurses. AIM: To understand the impact of the work environment on the emotional health of doctors and nurses in a general hospital setting. METHODS: Cross sectional study using self-reported questionnaires including the General Health Questionnaire, the Posttraumatic Stress Disorder (PTSD) Checklist for Civilians, the Trauma Experiences and Work Environment Scale. RESULTS: The response rates for the study were 28% (60) for doctors and 54% (431) for nurses. Whilst the prevalences of psychiatric disorder, anxiety, depression and PTSD were higher for doctors compared with nurses, this was not statistically significant. Both groups reported witnessing someone badly injured or killed as their most distressing experience (doctors 46% versus nurses 41%). Using multiple logistic regression, significant predictors of emotional health was task orientation for doctors (OR = 1.96, 95% CI = 1.1-3.6), and PTSD (OR = 17.2, 95% CI = 6.0-49.6), work pressure (OR = 1.2, 95% CI = 1.01-1.4) and innovation (OR = 0.81, 95% CI = 0.70-0.94) for nurses. CONCLUSION: The prevalence of psychiatric disorder among the doctors and nurses was similar to that in Britain. Elements of the work environment did impact on the emotional health of health care workers. Organizational development initiatives should include employee mental health issues in order to create a more positive work environment. PMID- 15133147 TI - Bacteriophage capsids: tough nanoshells with complex elastic properties. AB - The shell of bacteriophages protects the viral DNA during host-to-host transfer and serves as a high-pressure container storing energy for DNA injection into a host bacterium. Here, we probe the mechanical properties of nanometer-sized bacteriophage phi 29 shells by applying point forces. We show that empty shells withstand nanonewton forces while being indented up to 30% of their height. The elastic response varies across the surface, reflecting the arrangement of shell proteins. The measured Young's modulus (approximately 1.8 GPa) is comparable with that of hard plastic. We also observe fatigue and breakage of capsids after probing them repetitively. These results illustrate the mechanoprotection that viral shells provide and also suggest design principles for nanotechnology. PMID- 15133148 TI - Expression of phosphoenolpyruvate carboxylase and phosphoenolpyruvate carboxylase kinase genes. Implications for genotypic capacity and phenotypic plasticity in the expression of crassulacean acid metabolism. AB - In plants with crassulacean acid metabolism (CAM), dark CO2 uptake is mediated by phosphoenolpyruvate carboxylase (PEPC), an enzyme that can be regulated at transcriptional and posttranslational levels. Reversible phosphorylation of PEPC is catalyzed by a dedicated PEPC kinase, which in turn is regulated at the transcriptional level over the 24-h cycle in CAM plants. PEPC kinase controls the day/night regulation of PEPC during the CAM cycle, thus facilitating plasticity for optimizing CO2 uptake under different environmental conditions. To understand the importance of PEPC kinase in relation to its target PEPC in terms of CAM performance, the expression of the genes encoding the two enzymes was investigated in four species of Clusia that have photosynthetic patterns ranging from C3 photosynthesis to constitutive CAM. By linking changes in the expression of PEPC and PEPC kinase to day/night patterns of leaf gas exchange, organic acid, and soluble sugar contents under different environmental conditions, the genetic and metabolic limitations to CAM plasticity were assessed. The results indicate that PEPC expression is a major factor underpinning the genotypic capacity for CAM and that PEPC kinase expression does not appear to limit CAM. The day/night regulation of Ppck transcript abundance was found to be a consequence of CAM and the day/night cycling of associated metabolites, rather than the primary controlling factor for the temporal separation of carboxylation processes. PMID- 15133149 TI - CHLOROPLAST BIOGENESIS genes act cell and noncell autonomously in early chloroplast development. AB - In order to identify nuclear genes required for early chloroplast development, a collection of photosynthetic pigment mutants of Arabidopsis was assembled and screened for lines with extremely low levels of chlorophyll. Nine chloroplast biogenesis (clb) mutants that affect proplastid growth and thylakoid membrane formation and result in an albino seedling phenotype were identified. These mutations identify six new genes as well as a novel allele of cla1. clb mutants have less than 2% of wild-type chlorophyll levels, and little or no expression of nuclear and plastid-encoded genes required for chloroplast development and function. In all but one mutant, proplastids do not differentiate enough to form elongated stroma thylakoid membranes. Analysis of mutants during embryogenesis allows differentiation between CLB genes that act noncell autonomously, where partial maternal complementation of chloroplast development is observed in embryos, and those that act cell autonomously, where complementation during embryogenesis is not observed. Molecular characterization of the noncell autonomous clb4 mutant established that the CLB4 gene encodes for hydroxy-2 methyl-2-(E)-butenyl 4-diphosphate synthase (HDS), the next to the last enzyme of the methylerythritol 4-phosphate (MEP) pathway for the synthesis of plastidic isoprenoids. The noncell autonomous nature of the clb4 mutant suggests that products of the MEP pathway can travel between tissues, and provides in vivo evidence that some movement of MEP intermediates exists from the cytoplasm to the plastid. The isolation and characterization of clb mutants represents the first systematic study of genes required for early chloroplast development in Arabidopsis. PMID- 15133150 TI - Consistency of Nicotiana attenuata's herbivore- and jasmonate-induced transcriptional responses in the allotetraploid species Nicotiana quadrivalvis and Nicotiana clevelandii. AB - We examined the consistency of the native diploid Nicotiana attenuata (Na)'s herbivore-induced transcriptional changes in the two allotetraploid natives, Nicotiana clevelandii (Nc) and Nicotiana quadrivalvis (Nq), which are thought to be derived from hybridizations with an ancestral Na. An analysis of nuclear encoded chloroplast-expressed Gln synthetase gene (ncpGS) sequences found strong similarity between Nc and Na and between N. trigonophylla and the two allopolyploids. All species were elicited with methyl jasmonate (MeJA), or were wounded and treated with either water, Manduca sexta oral secretions and regurgitant (R), or the two most abundant fatty acid amino acid conjugates (F) in R to simulate herbivory. The induced transcriptional responses in all three species were compared with a cDNA microarray enriched in Na genes. Na had the fastest transcriptional responses followed by Nc and then Nq. Na's R- and F elicited responses were more similar to those from Nq, while the MeJA- or wound elicited responses were more consistent in Nc. Treatment of wounds with the full cocktail of elicitors found in R elicits more complex responses than does treatment with F. The species differ in their elicited JA responses, and these differences are mirrored in the expression of oxylipin genes (LOX, HPL, AOS, and alpha-DOX) and downstream JA-elicited genes (TD). Elicitation decreases the expression of growth-related genes in all three species. We propose that this is a valuable system to examine the modification of complex, polygenic, adaptive responses during allopolyploid speciation. PMID- 15133151 TI - Characterization of GaWRKY1, a cotton transcription factor that regulates the sesquiterpene synthase gene (+)-delta-cadinene synthase-A. AB - The cotton (+)-delta-cadinene synthase (CAD1), a sesquiterpene cyclase, catalyzes a branch-point step leading to biosynthesis of sesquiterpene phytoalexins, including gossypol. CAD1-A is a member of CAD1 gene family, and its promoter contains a W-box palindrome with two reversely oriented TGAC repeats, which are the proposed binding sites of WRKY transcription factors. We isolated several WRKY cDNAs from Gossypium arboreum. One of them, GaWRKY1, encodes a protein containing a single WRKY domain and a putative N-terminal Leu zipper. Similar to genes encoding enzymes of cotton sesquiterpene pathway, GaWRKY1 was down regulated in a glandless cotton cultivar that contained much less gossypol. GaWRKY1 showed a temporal and spatial pattern of expression comparable to that of CAD1-A in various aerial organs examined, including sepal, stigma, anther, and developing seeds. In suspension cells, expression of both GaWRKY1 and CAD1-A genes and biosynthesis of sesquiterpene aldehydes were strongly induced by a fungal elicitor preparation and methyl jasmonate. GaWRKY1 interacted with the 3x W-box derived from CAD1-A promoter in yeast (Saccharomyces cerevisiae) one-hybrid system and in vitro. Furthermore, in transgenic Arabidopsis plants, overexpression of GaWRKY1 highly activated the CAD1-A promoter, and transient assay in tobacco (Nicotiana tabacum) leaves demonstrated that W-box was required for this activation. These results suggest that GaWRKY1 participates in regulation of sesquiterpene biosynthesis in cotton, and CAD1-A is a target gene of this transcription factor. PMID- 15133152 TI - The control of storage xyloglucan mobilization in cotyledons of Hymenaea courbaril. AB - Hymenaea courbaril is a leguminous tree species from the neotropical rain forests. Its cotyledons are largely enriched with a storage cell wall polysaccharide (xyloglucan). Studies of cell wall storage polymers have been focused mostly on the mechanisms of their disassembly, whereas the control of their mobilization and the relationship between their metabolism and seedling development is not well understood. Here, we show that xyloglucan mobilization is strictly controlled by the development of first leaves of the seedling, with the start of its degradation occurring after the beginning of eophyll (first leaves) expansion. During the period of storage mobilization, an increase in the levels of xyloglucan hydrolases, starch, and free sugars were observed in the cotyledons. Xyloglucan mobilization was inhibited by shoot excision, darkness, and by treatment with the auxin-transport inhibitor N-1-naphthylphthalamic acid. Analyses of endogenous indole-3-acetic acid in the cotyledons revealed that its increase in concentration is followed by the rise in xyloglucan hydrolase activities, indicating that auxin is directly related to xyloglucan mobilization. Cotyledons detached during xyloglucan mobilization and treated with 2,4 dichlorophenoxyacetic acid showed a similar mobilization rate as in attached cotyledons. This hormonal control is probably essential for the ecophysiological performance of this species in their natural environment since it is the main factor responsible for promoting synchronism between shoot growth and reserve degradation. This is likely to increase the efficiency of carbon reserves utilization by the growing seedling in the understorey light conditions of the rain forest. PMID- 15133153 TI - Nitrogen supply influences herbivore-induced direct and indirect defenses and transcriptional responses in Nicotiana attenuata. AB - Although nitrogen (N) availability is known to alter constitutive resistance against herbivores, its influence on herbivore-induced responses, including signaling pathways, transcriptional signatures, and the subsequently elicited chemical defenses is poorly understood. We used the native tobacco, Nicotiana attenuata, which germinates in the postfire environment and copes with large changes in soil N during postfire succession, to compare a suite of Manduca sexta and elicitor-induced responses in plants grown under high- and low-N (LN) supply rates. LN supply decreased relative growth rates and biomass by 35% at 40 d compared to high-N plants; furthermore, it also attenuated (by 39 and 60%) the elicitor-induced jasmonate and salicylate bursts, two N-intensive direct defenses (nicotine and trypsin proteinase inhibitors, albeit by different mechanisms), and carbon-containing nonvolatile defenses (rutin, chlorogenic acid, and diterpene glycosides), but did not affect the induced release of volatiles (cis-alpha bergamotene and germacrene A), which function as indirect defenses. M. sexta and methyl jasmonate-induced transcriptional responses measured with a microarray enriched in herbivore-induced genes were also substantially reduced in plants grown under LN supply rates. In M. sexta-attacked LN plants, only 36 (45%) up regulated and 46 (58%) down-regulated genes showed the same regulation as those in attacked high-N plants. However, transcriptional responses frequently directly countered the observed metabolic changes. Changes in a leaf's sensitivity to elicitation, an attacked leaf's waning ability to export oxylipin wound signals, and/or resource limitations in LN plants can account for the observed results, underscoring the conclusion that defense activation is a resource-intensive response. PMID- 15133154 TI - Molecular aspects of plant adaptation to life in the Chernobyl zone. AB - With each passing year since the Chernobyl accident of 1986, more questions arise about the potential for organisms to adapt to radiation exposure. Often this is thought to be attributed to somatic and germline mutation rates in various organisms. We analyzed the adaptability of native Arabidopsis plants collected from areas with different levels of contamination around the Chernobyl nuclear power plant from 1986 to 1992. Notably, progeny of Chernobyl plants resisted higher concentrations of the mutagens Rose Bengal and methyl methane sulfonate. We analyzed the possible molecular mechanisms of their resistance to mutagens and found a more than 10-fold lower frequency of extrachromosomal homologous recombination, significant differences in the expression of radical scavenging (CAT1 and FSD3) and DNA-repair (RAD1 and RAD51-like) genes upon exposure to mutagens (Rose Bengal and x-rays), and a higher level of global genome methylation. This data suggests that adaptation to ionizing radiation is a complex process involving epigenetic regulation of gene expression and genome stabilization that improves plants' resistance to environmental mutagens. PMID- 15133155 TI - Metabolic fate of jasmonates in tobacco bright yellow-2 cells. AB - Jasmonic acid and methyl jasmonate play an essential role in plant defense responses and pollen development. Their levels are temporarily and spatially controlled in plant tissue. However, whereas jasmonate biosynthesis is well studied, metabolic pathways downstream of jasmonic acid are less understood. We studied the uptake and metabolism of jasmonic acid and methyl jasmonate in tobacco (Nicotiana tabacum) Bright Yellow-2 suspension culture. We found that upon uptake, jasmonic acid was metabolized to its Glc and gentiobiose esters, and hydroxylation at C-11 or C-12 occurred. Free hydroxylated jasmonates were the preferential fraction of the culture medium. Upon hydrolysis of methyl jasmonate to jasmonic acid, a similar set of conversions occurs. In contrast to jasmonic acid, none of its derivatives interfere with the G2/M transition in synchronized tobacco Bright Yellow-2 cells. PMID- 15133156 TI - Crop improvement through modification of the plant's own genome. AB - Plant genetic engineering has, until now, relied on the incorporation of foreign DNA into plant genomes. Public concern about the extent to which transgenic crops differ from their traditionally bred counterparts has resulted in molecular strategies and gene choices that limit, but not eliminate, the introduction of foreign DNA. Here, we demonstrate that a plant-derived (P-) DNA fragment can be used to replace the universally employed Agrobacterium transfer (T-) DNA. Marker free P-DNAs are transferred to plant cell nuclei together with conventional T DNAs carrying a selectable marker gene. By subsequently linking a positive selection for temporary marker gene expression to a negative selection against marker gene integration, 29% of derived regeneration events contain P-DNA insertions but lack any copies of the T-DNA. Further refinements are accomplished by employing Omega-mutated virD2 and isopentenyl transferase cytokinin genes to impair T-DNA integration and select against backbone integration, respectively. The presented methods are used to produce hundreds of marker-free and backbone free potato (Solanum tuberosum) plants displaying reduced expression of a tuber specific polyphenol oxidase gene in potato. The modified plants represent the first example of genetically engineered plants that only contain native DNA. PMID- 15133157 TI - The role of the jasmonate response in plant susceptibility to diverse pathogens with a range of lifestyles. AB - Plants defend themselves against attack from insects and pathogens with various resistance strategies. The jasmonate and salicylate signaling pathways are two induced responses that protect plants against these attackers. Knowledge of the range of organisms that are affected by each response is important for understanding how plants coordinate their defenses against multiple attackers and the generality of effect of different resistance mechanisms. The jasmonate response is known to protect plants against a wide range of insect herbivores; in this study, we examined the role of the jasmonate response in susceptibility to eight pathogens with diverse lifestyles in the laboratory and field. Recent biochemical models suggest that the lifestyle of the pathogen (necrotroph versus biotroph) should predict whether the jasmonate response will be involved in resistance. We tested this by examining the susceptibility of wild-type (cv Castlemart with no known genes for resistance to the pathogens used) and jasmonate-deficient mutant tomato (Lycopersicon esculentum) plants (def1) and by employing rescue treatments of the mutant. Plant susceptibility to five of the eight pathogens we examined was reduced by the jasmonate response, including two bacteria (Pseudomonas syringae and Xanthomonas campestris), two fungi (Verticillium dahliae and Fusarium oxysporum f. sp. lycopersici), and an oomycete (Phytophthora infestans). Susceptibility to three fungi was unaffected (Cladosporium fulvum, Oidium neolycopersici, and Septoria lycopersici). Our results indicate that the jasmonate response reduces damage by a wide range of pathogens from different lifestyles, a result that contrasts with the emerging picture of diseases on Arabidopsis. Thus, the generality of jasmonate-based resistance of tomato challenges the view that ecologically distinct plant parasites are resisted via different mechanisms. PMID- 15133158 TI - Identification of the catalytic motif of the microbial ribosome inactivating cytotoxin colicin E3. AB - Colicin E3 is a cytotoxic ribonuclease that specifically cleaves 16S rRNA at the ribosomal A-site to abolish protein synthesis in sensitive Escherichia coli cells. We have performed extensive mutagenesis of the 96-residue colicin E3 cytotoxic domain (E3 rRNase), assayed mutant colicins for in vivo cytotoxicity, and tested the corresponding E3 rRNase domains for their ability to inactivate ribosome function in vitro. From 21 alanine mutants, we identified five positions where mutation resulted in a colicin with no measurable cytotoxicity (Y52, D55, H58, E62, and Y64) and four positions (R40, R42, E60, and R90) where mutation caused a significant reduction in cytotoxicity. Mutations that were found to have large in vivo and in vitro effects were tested for structural integrity through circular dichroism and fluorescence spectroscopy using purified rRNase domains. Our data indicate that H58 and E62 likely act as the acid-base pair during catalysis with other residues likely involved in transition state stabilization. Both the Y52 and Y64 mutants were found to be highly destabilized and this is the likely origin of the loss of their cytotoxicity. The identification of important active site residues and sequence alignments of known rRNase homologs has allowed us to identify other proteins containing the putative rRNase active site motif. Proteins that contained this active site motif included three hemagglutinin-type adhesins and we speculate that these have evolved to deliver a cytotoxic rRNase into eukaryotic cells during pathogenesis. PMID- 15133159 TI - Accessibility of introduced cysteines in chemoreceptor transmembrane helices reveals boundaries interior to bracketing charged residues. AB - Two hydrophobic sequences, 24 and 30 residues long, identify the membrane spanning segments of chemoreceptor Trg from Escherichia coli. As in other related chemoreceptors, these helical sequences are longer than the minimum necessary for an alpha-helix to span the hydrocarbon region of a biological membrane. Thus, the specific positioning of the segments relative to the hydrophobic part of the membrane cannot be deduced from sequence alone. With the aim of defining the positioning for Trg experimentally, we determined accessibility of a hydrophilic sulfhydryl reagent to cysteines introduced at each position within and immediately outside the two hydrophobic sequences. For both sequences, there was a specific region of uniformly low accessibility, bracketed by regions of substantial accessibility. The two low-accessibility regions were each 19 residues long and were in register in the three-dimensional organization of the transmembrane domain deduced from independent data. None of the four hydrophobic hydrophilic boundaries for these two membrane-embedded sequences occurred at a charged residue. Instead, they were displaced one to seven residues internal to the charged side chains bracketing the extended hydrophobic sequences. Many hydrophobic sequences, known or predicted to be membrane-spanning, are longer than the minimum necessary helical length, but precise membrane boundaries are known for very few. The cysteine-accessibility approach provides an experimental strategy for determining those boundaries that could be widely applicable. PMID- 15133160 TI - Two-promoter vector is highly efficient for overproduction of protein complexes. AB - The use of bicistronic vectors, which contain two target genes under one promoter, has been the most common practice for the heterologous production of binary protein complexes. The major problem of this method is the much lower expression of the second gene compared with that of the first gene next to the promoter. We tested a simple idea of whether inclusion of an additional promoter in front of the second gene may remove the problem. Compared with bicistronic vectors, corresponding two-promoter vectors yielded four to nine times larger amounts of the complexes between BCL-2 family proteins, BCL-X(L):BAD, BCL X(L):BIM-S, and CED-9:EGL-1 in bacterial cells as a result of significantly increased expression of the second genes in a manner independent of the order of the target genes. With the two-promoter system, we produced two other complexes in large quantity suitable for extensive crystallization trial. The method does not accompany any technical disadvantages, and represents a significant improvement from the conventional method, which should enjoy wide application for the coexpression of binary or higher order protein complexes by extension. PMID- 15133161 TI - Sequence-structure mapping errors in the PDB: OB-fold domains. AB - The Protein Data Bank (PDB) is the single most important repository of structural data for proteins and other biologically relevant molecules. Therefore, it is critically important to keep the PDB data, as much as possible, error-free. In this study, we have analyzed PDB crystal structures possessing oligonucleotide/oligosaccharide binding (OB)-fold, one of the highly populated folds, for the presence of sequence-structure mapping errors. Using energy-based structure quality assessment coupled with sequence analyses, we have found that there are at least five OB-structures in the PDB that have regions where sequences have been incorrectly mapped onto the structure. We have demonstrated that the combination of these computation techniques is effective not only in detecting sequence-structure mapping errors, but also in providing guidance to correct them. Namely, we have used results of computational analysis to direct a revision of X-ray data for one of the PDB entries containing a fairly inconspicuous sequence-structure mapping error. The revised structure has been deposited with the PDB. We suggest use of computational energy assessment and sequence analysis techniques to facilitate structure determination when homologs having known structure are available to use as a reference. Such computational analysis may be useful in either guiding the sequence-structure assignment process or verifying the sequence mapping within poorly defined regions. PMID- 15133162 TI - Structure of a novel c7-type three-heme cytochrome domain from a multidomain cytochrome c polymer. AB - The structure of a novel c(7)-type cytochrome domain that has two bishistidine coordinated hemes and one heme with histidine, methionine coordination (where the sixth ligand is a methionine residue) was determined at 1.7 A resolution. This domain is a representative of domains that form three polymers encoded by the Geobacter sulfurreducens genome. Two of these polymers consist of four and one protein of nine c(7)-type domains with a total of 12 and 27 hemes, respectively. Four individual domains (termed A, B, C, and D) from one such multiheme cytochrome c (ORF03300) were cloned and expressed in Escherichia coli. The domain C produced diffraction quality crystals from 2.4 M sodium malonate (pH 7). The structure was solved by MAD method and refined to an R-factor of 19.5% and R-free of 21.8%. Unlike the two c(7) molecules with known structures, one from G. sulfurreducens (PpcA) and one from Desulfuromonas acetoxidans where all three hemes are bishistidine coordinated, this domain contains a heme which is coordinated by a methionine and a histidine residue. As a result, the corresponding heme could have a higher potential than the other two hemes. The apparent midpoint reduction potential, E(app), of domain C is -105 mV, 50 mV higher than that of PpcA. PMID- 15133163 TI - Pressure dissociation studies provide insight into oligomerization competence of temperature-sensitive folding mutants of P22 tailspike. AB - Several temperature-sensitive folding (tsf) mutants of the tailspike protein from bacteriophage P22 have been found to fold with lower efficiency than the wild type sequence, even at lowered temperatures. Previous refolding studies initiated from the unfolded monomer have indicated that the tsf mutations decrease the rate of structured monomer formation. We demonstrate that pressure treatment of the tailspike aggregates provides a useful tool to explore the effects of tsf mutants on the assembly pathway of the P22 tailspike trimer. The effects of pressure on two different tsf mutants, G244R and E196K, were explored. Pressure treatment of both G244R and E196K aggregates produced a folded trimer. E196K forms almost no native trimer in in vitro refolding experiments, yet it forms a trimer following pressure in a manner similar to the native tailspike protein. In contrast, trimer formation from pressure-treated G244R aggregates was not rapid, despite the presence of a G244R dimer after pressure treatment. The center-of-mass shifts of the fluorescence spectra under pressure are nearly identical for both tsf aggregates, indicating that pressure generates similar intermediates. Taken together, these results suggest that E196K has a primary defect in formation of the beta-helix during monomer collapse, while G244R is primarily an assembly defect. PMID- 15133164 TI - Activation and inhibition of cyclin-dependent kinase-2 by phosphorylation; a molecular dynamics study reveals the functional importance of the glycine-rich loop. AB - Nanoseconds long molecular dynamics (MD) trajectories of differently active complexes of human cyclin-dependent kinase 2 (inactive CDK2/ATP, semiactive CDK2/Cyclin A/ATP, fully active pT160-CDK2/Cyclin A/ATP, inhibited pT14-; pY15-; and pT14,pY15,pT160-CDK2/Cyclin A/ATP) were compared. The MD simulations results of CDK2 inhibition by phosphorylation at T14 and/or Y15 sites provide insight into the structural aspects of CDK2 deactivation. The inhibitory sites are localized in the glycine-rich loop (G-loop) positioned opposite the activation T loop. Phosphorylation of T14 and both inhibitory sites T14 and Y15 together causes ATP misalignment for phosphorylation and G-loop conformational change. This conformational change leads to the opening of the CDK2 substrate binding box. The phosphorylated Y15 residue negatively affects substrate binding or its correct alignment for ATP terminal phospho-group transfer to the CDK2 substrate. The MD simulations of the CDK2 activation process provide results in agreement with previous X-ray data. PMID- 15133165 TI - Realistic protein-protein association rates from a simple diffusional model neglecting long-range interactions, free energy barriers, and landscape ruggedness. AB - We develop a simple but rigorous model of protein-protein association kinetics based on diffusional association on free energy landscapes obtained by sampling configurations within and surrounding the native complex binding funnels. Guided by results obtained on exactly solvable model problems, we transform the problem of diffusion in a potential into free diffusion in the presence of an absorbing zone spanning the entrance to the binding funnel. The free diffusion problem is solved using a recently derived analytic expression for the rate of association of asymmetrically oriented molecules. Despite the required high steric specificity and the absence of long-range attractive interactions, the computed rates are typically on the order of 10(4)-10(6) M(-1) sec(-1), several orders of magnitude higher than rates obtained using a purely probabilistic model in which the association rate for free diffusion of uniformly reactive molecules is multiplied by the probability of a correct alignment of the two partners in a random collision. As the association rates of many protein-protein complexes are also in the 10(5)-10(6) M(-1) sec(-1) range, our results suggest that free energy barriers arising from desolvation and/or side-chain freezing during complex formation or increased ruggedness within the binding funnel, which are completely neglected in our simple diffusional model, do not contribute significantly to the dynamics of protein-protein association. The transparent physical interpretation of our approach that computes association rates directly from the size and geometry of protein-protein binding funnels makes it a useful complement to Brownian dynamics simulations. PMID- 15133166 TI - Modification of halogen specificity of a vanadium-dependent bromoperoxidase. AB - The halide specificity of vanadium-dependent bromoperoxidase (BPO) from the marine algae, Corallina pilulifera, has been changed by a single amino acid substitution. The residue R397 has been substituted by the other 19 amino acids. The mutant enzymes R397W and R397F showed significant chloroperoxidase (CPO) activity as well as BPO activity. These mutant enzymes were purified and their properties were investigated. The maximal velocities of CPO activities of the R397W and R397F enzymes were 31.2 and 39.2 units/mg, and the K(m) values for Cl( ) were 780 mM and 670 mM, respectively. Unlike the native enzyme, both mutant enzymes were inhibited by NaN(3). In the case of the R397W enzyme, the incorporation rate of vanadate into the active site was low, compared with the R397F and the wild-type enzyme. These results supported the existence of a specific halogen binding site within the catalytic cleft of vanadium haloperoxidases. PMID- 15133167 TI - Production of a fully functional, permuted single-chain penicillin G acylase. AB - Penicillin G acylase (PGA) is a heterodimeric enzyme synthesized as a single polypeptide precursor that undergoes an autocatalytic processing to remove an internal spacer peptide to produce the active enzyme. We constructed a single chain PGA not dependent on autoproteolytic processing. The mature sequence of the beta-domain was expressed as the N terminus of a new polypeptide, connected by a random tetra-peptide to the alpha-domain, to afford a permuted protein. We found several active enzymes among variants differing in their linker peptides. Protein expression analysis showed that the functional single-chain variants were produced when using a Sec-dependent leader peptide, or when expressed inside the bacterial cytoplasm. Active-site titration experiments showed that the single chain proteins displayed similar k(cat) values to the ones obtained with the wild type enzyme. Interestingly, the single-chain proteins also displayed close to 100% of functional active sites compared to 40% to 70% functional yield usually obtained with the heterodimeric protein. PMID- 15133168 TI - p53 expression and environmental tobacco smoke exposure in feline oral squamous cell carcinoma. AB - The purpose of this study was to determine the prevalence of p53 overexpression in feline oral squamous cell carcinomas (SCC) and to determine, if any, the association between p53 overexpression and lifestyle factors and environmental exposures, including exposure to environmental tobacco smoke (ETS). Questionnaires concerning exposure to ETS and other environmental factors were sent to owners of cats presenting to the Harrington Oncology Program with a diagnosis of oral SCC between 1991 and 2000. Additionally, 23 formalin-fixed biopsy samples from these cats, with information regarding ETS, were evaluated immunohistochemically for p53 expression using the CM-1 clone and the avidin biotin-horseradish peroxidase method. Of the 23 samples evaluated, 15 (65%) showed positive nuclear staining for the CM-1 clone. Tumor biopsy samples from cats exposed to any ETS were 4.5 times more likely to overexpress p53 than were tumors from unexposed cats (P = 0.19). Among cats with any ETS exposure, those with 5 years or longer of exposure were 7.0 times more likely to overexpress p53 (P = 0.38). Longhaired cats (P = 0.18) and female cats (P = 0.35) were also more likely to show p53 expression in their tumors. These results provide additional support for a relationship between oral SCC development and exposure to household ETS and may implicate p53 as a potential site for carcinogen-related mutation in this tumor. PMID- 15133169 TI - Summer pheasant's eye (Adonis aestivalis) poisoning in three horses. AB - Three horses died as a result of eating grass hay containing summer pheasant's eye (Adonis aestivalis L.), a plant containing cardenolides similar to oleander and foxglove. A 9-year-old thoroughbred gelding, a 20-year-old appaloosa gelding, and a 5-year-old quarter horse gelding initially presented with signs of colic 24 48 hours after first exposure to the hay. Gastrointestinal gaseous distension was the primary finding on clinical examination of all three horses. Two horses became moribund and were euthanatized 1 day after first showing clinical signs, and the third horse was euthanatized after 4 days of medical therapy. Endocardial hemorrhage and gaseous distension of the gastrointestinal tract were the only necropsy findings in the first two horses. On microscopic examination, both horses had scattered foci of mild, acute myocardial necrosis and neutrophilic inflammation associated with endocardial and epicardial hemorrhage. The third horse that survived for 4 days had multifocal to coalescing, irregular foci of acute, subacute, and chronic myocardial degeneration and necrosis. A. aestivalis (pheasant's eye, summer adonis) was identified in the hay. Strophanthidin, the aglycone of several cardenolides present in Adonis spp., was detected by liquid chromatography-mass spectrometry-mass spectrometry in gastrointestinal contents from all three horses. Although Adonis spp. contain cardiac glycosides, cardiac lesions have not previously been described in livestock associated with consumption of adonis, and this is the first report of adonis toxicosis in North America. PMID- 15133170 TI - A von Willebrand's factor genomic nucleotide variant and polymerase chain reaction diagnostic test associated with inheritable type-2 von Willebrand's disease in a line of german shorthaired pointer dogs. AB - Heritable, type-2 von Willebrand's disease (vWD) was studied in a line of German Shorthaired Pointers (GSPs) in which some members had a nucleotide variant in exon 28 of the von Willebrand factor (VWF) gene. A polymerase chain reaction (PCR) diagnostic test for the nucleotide variant was developed to establish the disorder's mode of inheritance and to eliminate it from the line. Thirty-six of the 49 GSPs in the line, 14 unrelated GSP controls, and 71 unrelated dogs of various breeds were tested for the presence of the variant nucleotide. All the dogs with a vWF antigen deficiency (<70% of normal) were either homozygous or heterozygous for the nucleotide variant. The variant was not located in any tested dog in the line or outside of the line with a vWF antigen value greater than 68%. Of the GSPs in the line tested, two were homozygous for the variant, 15 were heterozygous, and 19 were variant free. The collective evidence of this and other studies is consistent with the variant nucleotide being the cause of the type-2 vWD in this line of GSPs and German Wirehaired Pointers. The PCR diagnostic test for the variant nucleotide was successfully used to select and produce progeny that were variant free and vWD free. This test should be effective in the subsequent elimination of this same variant from other lines of dogs. PMID- 15133171 TI - Hepatic alveolar echinococcosis in cynomolgus monkeys (Macaca fascicularis). AB - Alveolar echinococcosis was diagnosed in 12 cynomolgus monkeys (Macaca fascicularis) at postmortem examination within a period of 6 years. Besides consistent involvement of the liver, parasitic lesions were also present in mesenteric lymph nodes, pancreas, lung, and kidney. In the liver, various patterns of host's responses to parasitic tissue could be distinguished. Infiltration of macrophages, often multinucleated, around usually intact metacestodes was the main feature of one pattern. A second pattern was characterized by the presence of abundant, normally degenerate granulocytes in addition to macrophages surrounding collapsed laminated structures. Finally and as a third pattern, some cysts were surrounded by marked collagen deposition, which was usually not a significant feature of the other foci. Parasitic cysts with protoscolices were observed in foci with the first and third pattern but not in the second one. The simultaneous occurrence of all three patterns was observed in most animals. Type AA amyloid was identified either in the space of Disse, macrophages or blood vessel walls in nine animals using immunohistochemistry. Identity of parasitic structures such as metacestodes of Echinococcus multilocularis was confirmed immunohistochemically. All animals that could be tested serologically (7/12) had detectable antibodies against the E. multilocularis-specific Em2 antigen. Liver lesions of six animals were additionally analyzed by polymerase chain reaction, yielding the amplification of a specific E. multilocularis DNA fragment in each case. PMID- 15133172 TI - Interleukin-1beta and tumor necrosis factor-alpha produce distinct, time dependent patterns of acute arthritis in the rat knee. AB - Interleukin-1beta (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha) synergistically induce and sustain arthritis. Two competing hypotheses of arthritis induction are 1) that TNF preferentially mediates inflammation, whereas IL-1 impels bone destruction, or 2) that either cytokine controls the entire process. In this study, these propositions were tested in two experiments by instilling IL-1beta or TNF-alpha into one knee of Lewis rats (n = 6/group) to incite arthritis, after which semiquantitative scores for inflammation, bone resorption, osteoclasts, and cartilage integrity were acquired. In the induction study, IL-1beta or TNF-alpha (3, 10, or 30 micro g) was given once to incite arthritis. After 2 days, IL-1beta induced significant, dose-dependent increases in inflammation (mild to marked), bone resorption (minimal to moderate), and osteoclasts (minimal to moderate). In contrast, TNF-alpha induced minimal to mild inflammation but had little impact on resorption or osteoclasts. Both IL-1 and TNF (>/=10 micro g) yielded mild cartilage degeneration. Most lesion scores in TNF-treated rats were significantly lower than those in animals given the same dose of IL-1beta. In the persistence study, rats were injected once with IL-1 or TNF (10 micro g) and maintained for 2, 3, or 7 days. IL-1beta significantly enhanced inflammation (all 3 days), bone resorption (days 2 and 3), osteoclasts (days 2 and 3), and cartilage matrix loss (days 2 and 3), whereas TNF-alpha augmented inflammation (days 2 and 3) and cartilage degeneration (day 2) but not bone resorption or osteoclasts. Thus, both IL-1beta and TNF-alpha can launch inflammation, but IL-1beta drives skeletal destruction. PMID- 15133173 TI - Histopathologic findings and classification of feline renal transplants. AB - Seventy-seven feline transplant kidney specimens, obtained from 1 to 3,183 days (9 years) after transplantation, were reevaluated histologically and classified on the basis of the Banff '97 guidelines for human renal transplant kidneys. Overall, this classification system appeared useful in detecting rejection reactions and confirmed the finding in humans that biopsies can diagnose subclinical rejection and therefore are an important diagnostic tool for the follow up of renal transplants. However, on the basis of serum creatinine values, the severity of the acute or active and chronic lesions was not accurately reflected by this scoring system. This is thought to be due to the significant differences in histologic rejection patterns, especially in acute or active rejection, in cats when compared with humans. Tubulitis, lymphocytic glomerulitis, and vasculitis, which are the main pillars of the Banff '97 acute or active rejection scoring system, are either rare or not found in cats. The presence of significant necrotizing glomerulitis and vasculitis in feline renal transplants might imply that the rejection is complicated by acute antibody mediated rejection. Alternatively, cyclosporine toxicity also should be considered because some of these kidneys show other signs of cyclosporine toxicity. Finally, the significance of subcapsular and interlobular phlebitis, rarely described in human rejection reactions but a distinct entity in cats, is unknown. From this study, it is clear that there are significant differences in the histology of acute or active rejection between humans and cats and that a better understanding of the histologic appearance of renal allografts will be especially beneficial for treatment and prognostic purposes. PMID- 15133174 TI - Pathologic, immunohistochemical, and electron microscopic findings in naturally occurring virulent systemic feline calicivirus infection in cats. AB - Infection with feline calicivirus (FCV) is a common cause of upper respiratory and oral disease in cats. FCV infection is rarely fatal, however, virulent, systemic strains of FCV (VS-FCV) that cause alopecia, cutaneous ulcers, subcutaneous edema, and high mortality in affected cats have recently been described. Seven cats with natural VS-FCV infection all had subcutaneous edema and ulceration of the oral cavity, with variable ulceration of the pinnae, pawpads, nares, and skin. Other lesions that were present in some affected cats included bronchointerstitial pneumonia, and pancreatic, hepatic, and splenic necrosis. Viral antigen was present within endothelial and epithelial cells in affected tissues as determined by immunohistochemical staining with a monoclonal antibody to FCV. Mature intranuclear and intracytoplasmic virions in necrotic epithelial cells were identified by transmission electron microscopy. VS-FCV infection causes epithelial cell cytolysis and systemic vascular compromise in susceptible cats, leading to cutaneous ulceration, severe edema, and high mortality. PMID- 15133175 TI - Multipotential osteosarcoma with various mesenchymal differentiations in a young dog. AB - Apparently synchronous, aggressive, mixed mesenchymal tumors in the right tibia, right femur, left femur, and rib cage produced multiple microscopic metastases in the lungs and macroscopic metastases in the liver, kidney, and spleen in a 1.5 year-old, neutered male, mixed-breed dog. No primary soft tissue tumor mass was present. Microscopically, the neoplasm exhibited osteosarcomatous, chondrosarcomatous, liposarcomatous, leiomyosarcomatous, fibrosarcomatous, angiosarcomatous, and leukocytic differentiation and was diagnosed as a multipotential osteosarcoma with various mesenchymal differentiation. Immunohistochemically, the neoplasm was cytoplasmically immunoreactive for vimentin, osteonectin, osteocalcin, CD 18, CD 31, desmin, and muscle-specific actin. Oil Red O staining was positive within liposarcomatous areas. Skeletal metastases from a primary bone tumor are exceedingly rare in human and veterinary medicine. However, the history, clinical signs, location, microscopic and immunohistochemical features were similar to those described in aggressive, poorly differentiated osteosarcomas of children. In addition, the wide range of mesenchymal tissue differentiation of this neoplasm was unusual, and to the authors' knowledge, an osteosarcoma with this degree of multiple differentiation has not been previously reported in the dog. PMID- 15133176 TI - Immunohistochemical expression of estrogen receptor beta in normal and tumoral canine mammary glands. AB - To date, two isoforms of estrogen receptors (ER) have been identified, cloned, and characterized from several species, estrogen receptor-alpha (ERalpha) and estrogen receptor-beta (ERbeta). Although the presence of ERalpha has been demonstrated in normal and tumoral canine mammary tissues, the issue of ERbeta expression has not been addressed in the dog. In this study, we have analyzed the expression of ERbeta in formalin-fixed, paraffin-embedded tissue samples of nonaltered mammary gland, 30 malignant (six complex carcinoma, 12 simple carcinoma, three carcinosarcoma, and nine carcinoma or sarcoma in benign tumor), and five benign (one fibroadenoma, one complex papilloma, one complex adenoma, and two benign mixed tumors) mammary tumors of the dog by using a polyclonal ERbeta antibody and the avidin-biotin-peroxidase complex immunohistochemical technique. Our results show that high numbers of normal ductal and acinar epithelium and approximately one third of canine mammary tumors express ERbeta. This expression was higher in benign than in malignant tumors. Furthermore, expression was higher in complex and mixed histologic subtypes of malignant tumors when compared with simple subtypes. PMID- 15133177 TI - Multifocal rhabdomyosarcomas within the tongue and oral cavity of a dog. AB - A 10-year-old terrier crossbreed presented with a change in bark intonation of 3 4 month's duration and pronounced panting. Four variably sized masses were observed within the oral cavity. The largest mass was located within the parenchyma at the caudal region of the tongue. Others were located on the left arytenoid, within the soft palate, and in the oropharynx above the soft palate. Histopathologic specimens consisted of large round to polygonal cells occasionally containing multiple nuclei and rare faint cytoplasmic cross striations. Staining was weakly positive with periodic acid-Schiff. Immunocytochemistry was strongly diffusely positive for muscle-specific actin, myoglobin, and desmin and scattered positive for S-100 and vimentin. Phosphotungstic acid-hematoxylin staining enhanced cytoplasmic cross striations. The cytoplasm of all neoplastic cells was filled with mitochondria on electron microscopy. The final diagnosis was multifocal/metastatic rhabdomyosarcoma. PMID- 15133178 TI - Expression of myogenic regulating factors, Myogenin and MyoD, in two canine botryoid rhabdomyosarcomas. AB - Myogenin and MyoD regulate the development of skeletal muscle, and their expressions are specific to the stages of myogenesis. Therefore, these myogenic regulatory proteins could be considered as sensitive and specific markers for rhabdomyosarcoma. In this report we investigated the immunohistochemical reactivities of myogenin and MyoD in two canine bladder botryoid rhabdomyosarcomas that were different in the degree of differentiation. MyoD was stained in the Ki-67 antigen-positive undifferentiated mesenchymal cells, which had proliferative activity similar to myoblasts differentiated from mesoblasts. In contrast, multinucleated neoplastic cells were positive for myogenin and alpha sarcomeric actin but not for Ki-67 antigen, similar to the myotubes differentiated from myoblastic cells. The expressions of myogenin and MyoD were closely correlated to the histologic features of myogenic neoplastic cells. PMID- 15133179 TI - The roles of Smad2 and Smad3 in the development of chemically induced skin tumors in mice. AB - Transforming growth factor-beta (TGF-beta) plays a complex role in skin carcinogenesis, acting as a suppressor early in tumor development but later as a promoter. Smad proteins are important intracellular mediators of TGF-beta signaling. To determine the effect of disrupting Smad genes and TGF-beta signaling on chemically induced skin carcinogenesis in mice, transgenic mice heterozygous for Smad2 or Smad3 deletions and wild-type controls were treated with topical dimethylbenzanthracene for 7 months. Tumor multiplicity, type, and degree of differentiation were assessed by histopathology and immunohistochemistry. Smad3(+/-) mice developed significantly fewer tumors than the wild-type group (P < 0.05). This indicated a possible oncogenic function for Smad3 in skin carcinogenesis. Smad2(+/-) mice formed less-differentiated tumors than their wild-type counterparts, supporting a tumor suppressor role for Smad2. There was a significant difference (P < 0.05) in tumor type between Smad2(+/-) and Smad3(+/-) groups, suggesting that Smad2 and Smad3 may regulate different targets. PMID- 15133180 TI - Cerebral ganglioneuroblastoma in a golden retriever dog. AB - An 8-month-old Golden Retriever dog was euthanatized because of a large cerebral mass extending from the right frontal lobe to the thalamus that was composed of both mature and immature neuronal cells. The better differentiated cells had abundant eosinophilic cytoplasm with prominent Nissl substance and were generally positive for neurofilament and variably positive for synaptophysin. The generally smaller and less-differentiated cells were infrequently positive for proliferating cell nuclear antigen and were negative for any neuronal and glial markers. No apparent glial differentiation of the immature tumor cells was detected. Based on morphologic and immunohistochemical features, the diagnosis of cerebral ganglioneuroblastoma was made. This neoplasm is very rare in all species, especially in the central nervous system, and has never been reported previously in this site in a dog. PMID- 15133181 TI - Nodular thyroid hyperplasia in a cynomolgus monkey. AB - Nodular thyroid hyperplasia was found in a 4-year-old male cynomolgus monkey (Macaca fascicularis). The monkey was clinically normal; however, necropsy revealed multiple variably sized nodules in both lobes of the thyroid gland. In contrast to the fairly uniform diameter of the lumen of follicles in the surrounding gland, the diameter of the follicular lumen within the hyperplastic nodules was highly variable and ranged from nonexistent to cystlike. Occasionally, in the larger follicles there were papillary infoldings of epithelium. The hyperplastic nodules were partially encapsulated by a fibrous capsule and showed little compression of the surrounding tissue. The follicular cells and colloid comprising the hyperplastic nodule were immunohistochemically positively stained with the antibody for thyroglobulin. Ultrastructurally, the cells forming follicles had numerous microvilli along the luminal surface, and lysosomal bodies and dilated rough endoplasmic reticulum in the cytoplasm. All these morphologic findings are consistent with nodular thyroid hyperplasia, which is rare in nonhuman primates. PMID- 15133182 TI - Malignant astrocytoma with binucleated granular cells in a Sprague-Dawley rat. AB - A 2-year-old Sprague-Dawley rat with hindlimb paralysis was diagnosed with a cerebral malignant astrocytoma. The distinctive feature of this astrocytoma was the presence of scattered binucleated cells that contained hypereosinophilic, 1-2 micro m in diameter, cytoplasmic granules. The neoplastic astrocytes stained positively for vimentin (VIM), lysozyme, and phosphotungstic acid hematoxylin (PTAH). Within the binucleated cells, granules stained with PTAH and periodic acid-Schiff (PAS) before and after diastase digestion. Ultrastructurally, neoplastic astrocytes were characterized by cytoplasmic aggregates of electron dense intermediate filaments consistent with VIM and desmin. The cytoplasm of binucleated cells contained numerous phagolysosomes enlarged by myelin figures and glycoprotein or glycolipid. Intermediate filaments were not present. This is the first description, in the rat, of a neoplasm with features resembling the human granular cell astrocytoma. Our findings suggest that an astrocytic origin should be considered for the binucleated cells in this neoplasm. PMID- 15133183 TI - Overexpression of the erbB-2 proto-oncogene in canine osteosarcoma cell lines and tumors. AB - The status of the erbB-2 (human epidermal growth factor receptor 2/neu) proto oncogene in canine osteosarcoma (OSA) has not been reported previously. In this study we used real-time reverse transcriptase polymerase chain reaction to evaluate erbB-2 expression in seven canine OSA cell lines and 10 canine OSA tissue samples. We determined erbB-2 to be significantly overexpressed in 86% (six of seven) of the cell lines and 40% (4 of 10) of the OSA tissues samples. Given the importance of erbB-2 in human breast cancer, the finding of erbB-2 overexpression in canine OSA may be important in further understanding the pathogenesis and possible therapies of OSA. PMID- 15133184 TI - Immunohistochemical expression of p27 and p21 in canine cutaneous mast cell tumors and histiocytomas. AB - The objective of this study was to evaluate by immunohistochemical means the nuclear expression of p27 and p21 proteins in cutaneous mast cell tumors and histiocytomas of dogs. In mast cell tumors, nine of the 13 grade I tumors, 13 of the 19 grade II tumors, and 10 of the 15 grade III tumors showed no detectable or mild p27 immunoreactivity. In contrast, one of the 13 grade I tumors, 12 of the 19 grade II tumors, and 11 of the 15 grade III tumors showed moderate or marked p21 immunoreactivity. Nineteen of the 28 histiocytomas showed no detectable or mild p27 immunoreactivity, and 24 cases showed moderate or marked p21 immunoreactivity. These findings indicate that a loss or absence of p27 expression is an early pathogenic event in mast cell and histiocyte tumorigenesis and that p21 expression may be a marker of mast cell tumor progression and histiocytoma cell proliferation. PMID- 15133185 TI - Acute bacterial cystitis does not cause deoxyribonucleic acid damage detectable by the alkaline comet assay in urothelial cells of dogs. AB - Considering the high incidence of dogs with acute bacterial cystitis (BC) and the relationship among inflammation, genotoxicity, and carcinogenesis, we conducted a case-control study comparing the frequency of deoxyribonucleic acid (DNA) lesions assessed by the comet assay between disease-free animals (13 males and 13 females) and cytology-confirmed cases of acute BC (12 males and 12 females), which was mainly caused by Staphylococcus sp. (40%) and Escherichia coli (35%). The results show no increase in DNA damage in cells obtained by bladder washings and no influence of age, sex, and breed due to acute BC. In conclusion, DNA damage was seemingly not associated with the infection by specific bacteria. PMID- 15133186 TI - Evolutionary genetics: the Drosophila model. PMID- 15133187 TI - Genetic variability of sexual size dimorphism in a natural population of Drosophila melanogaster: an isofemale-line approach. AB - Most animal species exhibit sexual size dimorphism (SSD). SSD is a trait difficult to quantify for genetical purposes since it must be simultaneously measured on two kinds of individuals, and it is generally expressed either as a difference or as a ratio between sexes. Here we ask two related questions: What is the best way to describe SSD, and is it possible to conveniently demonstrate its genetic variability in a natural population? We show that a simple experimental design, the isofemale-line technique (full-sib families), may provide an estimate of genetic variability, using the coefficient of intraclass correlation. We consider two SSD indices, the female-male difference and the female/male ratio. For two size-related traits, wing and thorax length, we found that both SSD indices were normally distributed. Within each family, the variability of SSD was estimated by considering individual values in one sex (the female) with respect to the mean value in the other sex (the male). In a homogeneous sample of 30 lines of Drosophila melanogaster, both indices provided similar intraclass correlations, on average 0.21, significantly greater than zero but lower than those for the traits themselves: 0.50 and 0.36 for wing and thorax length respectively. Wing and thorax length were strongly positively correlated within each sex. SSD indices of wing and thorax length were also positively correlated, but to a lesser degree than for the traits themselves. For comparative evolutionary studies, the ratio between sexes seems a better index of SSD since it avoids scaling effects among populations or species, permits comparisons between different traits, and has an unambiguous biological significance. In the case of D. melanogaster grown at 25 degrees C, the average female/male ratios are very similar for the wing (1.16) and the thorax (1.15), and indicate that, on average, these size traits are 15-16% longer in females. PMID- 15133188 TI - Mild heat stress at a young age in Drosophila melanogaster leads to increased Hsp70 synthesis after stress exposure later in life. AB - In a number of animal species it has been shown that exposure to low levels of stress at a young age has a positive effect on stress resistance later in life, and on longevity. The positive effects have been attributed to the activation of defence/cleaning systems (heat shock proteins (Hsps), antioxidases, DNA repair) or to effects of a changed metabolic rate, or both. We investigated the effect of mild stress exposures early in life on Hsp70 synthesis after a harder stress exposure later in life in five isofemale lines of Drosophila melanogaster. Female flies were either exposed to repeated bouts of mild heat stress (3 h at 34 degrees C) at a young age (days 2, 4 and 6 post-eclosion) or held under standard laboratory conditions. At 16 and 32 days of adult age, respectively, flies were exposed to a high-temperature treatment known to induce Hsp70 in the investigated species (1 h at 37 degrees C). Thereafter, the inducible Hsp70 levels were measured. Our data show a tendency towards increased Hsp70 synthesis with increased age for both 'mild stress' and 'no stress' flies. Moreover, the results show that flies exposed to mild stress at a young age synthesized more Hsp70 upon induction, compared to control flies, and that this difference was accentuated at 32 days compared to 16 days of age. Thus, bouts of mild heat stress at a young age impact on the physiological stress response system later in life. This may be caused by an increased ability to react to future stresses. Alternatively, the mild stress exposure at a young age may actually have caused cellular damages increasing the need for Hsp70 levels after stress exposure later in life. The importance of an Hsp70 upregulation (throughout life) in explaining the phenomenon of hormesis is discussed, together with alternative hypotheses, and suggestions for further studies. PMID- 15133189 TI - Quantitative-genetic analysis of wing form and bilateral asymmetry in isochromosomal lines of Drosophila subobscura using Procrustes methods. AB - Fluctuating asymmetry (FA) is often used as a measure of underlying developmental instability (DI), motivated by the idea that morphological variance is maladaptive. Whether or not DI has evolutionary potential is a highly disputed topic, marred by methodological problems and fuzzy prejudices. We report here some results from an ongoing study of the effects of karyotype, homozygosity and temperature on wing form and bilateral asymmetry using isochromosomal lines of Drosophila subobscura. Our approach uses the recently developed methodologies in geometric morphometrics to analyse shape configurations of landmarks within the standard statistical framework employed in studies of bilateral asymmetries, and we have extended these methods to partition the individual variation and the variation in asymmetries into genetic and environmental causal components. The analyses revealed temperature-dependent expression of genetic variation for wing size and wing shape, directional asymmetry (DA) of wing size, increased asymmetries at suboptimal temperature, and a transition from FA to DA in males as a result of increase in the rearing temperature. No genetic variation was generally detected for FA in our samples, but these are preliminary results because no crosses between lines were carried out and, therefore, the contribution of dominance was not taken into account. In addition, only a subset of the standing genetic variation was represented in the experiments. PMID- 15133191 TI - The devil in the details of life-history evolution: instability and reversal of genetic correlations during selection on Drosophila development. AB - The evolutionary relationships between three major components of Darwinian fitness, development rate, growth rate and preadult survival, were estimated using a comparison of 55 distinct populations of Drosophila melanogaster variously selected for age-specific fertility, environmental-stress tolerance and accelerated development. Development rate displayed a strong net negative evolutionary correlation with weight at eclosion across all selection treatments, consistent with the existence of a size-versus-time tradeoff between these characters. However, within the data set, the magnitude of the evolutionary correlation depended upon the particular selection treatments contrasted. A previously proposed tradeoff between preadult viability and growth rate was apparent only under weak selection for juvenile fitness components. Direct selection for rapid development led to sharp reductions in both growth rates and viability. These data add to the mounting results from experimental evolution that illustrate the sensitivity of evolutionary correlations to (i) genotype-by environment (G x E) interaction, (ii) complex functional-trait interactions, and (iii) character definition. Instability, disappearance and reversal of patterns of genetic covariation often occur over short evolutionary time frames and as the direct product of selection, rather than some stochastic process. We suggest that the functional architecture of fitness is a rapidly evolving matrix with reticulate properties, a matrix that we understand only poorly. PMID- 15133190 TI - Molecular population genetics of the beta-esterase gene cluster of Drosophila melanogaster. AB - We have investigated nucleotide polymorphism at the beta-esterase gene cluster including the Est-6 gene and psiEst-6 putative pseudogene in four samples of Drosophila melanogaster derived from natural populations of southern Africa (Zimbabwe), Europe (Spain), North America (USA: California), and South America (Venezuela). A complex haplotype structure is revealed in both Est-6 and psiEst 6. Total nucleotide diversity is twice in psiEst-6 as in Est-6; diversity is higher in the African sample than in the non-African ones. Strong linkage disequilibrium occurs within the beta-esterase gene cluster in non-African samples, but not in the African one. Intragenic gene conversion events are detected within Est-6 and, to a much greater extent, within psiEst-6; intergenic gene conversion events are rare. Tests of neutrality with recombination are significant for the beta-esterase gene cluster in the non-African samples but not significant in the African one. We suggest that the demographic history (bottleneck and admixture of genetically differentiated populations) is the major factor shaping the pattern of nucleotide polymorphism in the beta-esterase gene cluster. However there are some 'footprints' of directional and balancing selection shaping specific distribution of nucleotide polymorphism within the cluster. Intergenic epistatic selection between Est-6 and psiEst-6 may play an important role in the evolution of the beta-esterase gene cluster preserving the putative pseudogene from degenerative destruction and reflecting possible functional interaction between the functional gene and the putative pseudogene. Est-6 and psiEst-6 may represent an indivisible intergenic complex ('intergene') in which each single component (Est-6 or psiEst-6) cannot separately carry out the full functional role. PMID- 15133192 TI - The contribution of ancestry, chance, and past and ongoing selection to adaptive evolution. AB - The relative contributions of ancestry, chance, and past and ongoing selection to variation in one adaptive (larval feeding rate) and one seemingly nonadaptive (pupation height) trait were determined in populations of Drosophila melanogaster adapting to either low or high larval densities in the laboratory. Larval feeding rates increased rapidly in response to high density, and the effects of ancestry, past selection and chance were ameliorated by ongoing selection within 15-20 generations. Similarly, in populations previously kept at high larval density, and then switched to low larval density, the decline of larval feeding rate to ancestral levels was rapid (15-20 generations) and complete, providing support for a previously stated hypothesis regarding the costs of faster feeding in Drosophila larvae. Variation among individuals was the major contributor to variation in pupation height, a trait that would superficially appear to be nonadaptive in the environmental context of the populations used in this study because it did not diverge between sets of populations kept at low versus high larval density for many generations. However, the degree of divergence among populations (F(ST)) for pupation height was significantly less than expected for a selectively neutral trait, and we integrate results from previous studies to suggest that the variation for pupation height among populations is constrained by stabilizing selection, with a flat, plateau-like fitness function that, consequently, allows for substantial phenotypic variation within populations. Our results support the view that the genetic imprints of history (ancestry and past selection) in outbreeding sexual populations are typically likely to be transient in the face of ongoing selection and recombination. The results also illustrate the heuristic point that different forms of selection-for example directional versus stabilizing selection-acting on a trait in different populations may often not be due to differently shaped fitness functions, but rather due to differences in how the fitness function maps onto the actual distribution of phenotypes in a given population. We discuss these results in the light of previous work on reverse evolution, and the role of ancestry, chance, and past and ongoing selection in adaptive evolution. PMID- 15133193 TI - Hybridization, transgressive segregation and evolution of new genetic systems in Drosophila. AB - Introgressive hybridization facilitates incorporation of genes from one species into the gene pool of another. Studies on long-term effects of introgressive hybridization in animal systems are sparse. Drosophila nasuta (2n = 8) and D. albomicans (2n = 6)-a pair of allopatric, morphologically almost identical, cross fertile members of the nasuta subgroup of the immigrans species group-constitute an excellent system to analyse the impact of hybridization followed by transgressive segregation of parental characters in the hybrid progeny. Hybrid populations of D. nasuta and D. albomicans maintained for over 500 generations in the laboratory constitute new recombinant hybrid genomes, here termed cytoraces. The impact of hybridization, followed by introgression and transgressive segregation, on chromosomal constitution and karyotypes, some fitness parameters, isozymes, components of mating behaviour and mating preference reveals a complex pattern of interracial divergence among parental species and cytoraces. This assemblage of characters in different combinations in a laboratory hybrid zone allows us to study the emergence of new genetic systems. Here, we summarize results from our ongoing studies comparing these hybrid cytoraces with the parental species, and discuss the implications of these findings for our understanding of the evolution of new genetic systems. PMID- 15133194 TI - Thermal adaptation in Drosophila serrata under conditions linked to its southern border: unexpected patterns from laboratory selection suggest limited evolutionary potential. AB - To investigate the ability of Drosophila serrata to adapt to thermal conditions over winter at the species southern border, replicate lines from three source locations were held as discrete generations over three years at either 19 degrees C (40 generations) or temperatures fluctuating between 7 degrees C and 18 degrees C (20 generations). Populations in the fluctuating environment were maintained either with an adult 0 degrees C cold shock or without a shock. These conditions were expected to result in temperature-specific directional selection for increased viability and productivity under both temperature regimes, and reduced development time under the fluctuating-temperature regime. Selection responses of all lines were tested under both temperature regimes after controlling for carry over effects by rearing lines in these environments for two generations. When tested in the 19 degrees C environment, lines evolving at 19 degrees C showed a faster development time and a lower productivity relative to the other lines, while cold shock reduced development time and productivity of all lines. When tested in the fluctuating environment, productivity of the 7-18 degrees C lines selected with a cold shock was relatively lower than that of lines selected without a shock, but this pattern was not observed in the other populations. Viability and body size as measured by wing length were not altered by selection or cold shock, although there were consistent effects of source population on wing length. These results provide little evidence for temperature-specific adaptation in D. serrata-although the lines had diverged for some traits, these changes were not consistent with a priori predictions. In particular, there was no evidence for life-history changes reflecting adaptation to winter conditions at the southern border. The potential for D. serrata to adapt to winter conditions may therefore be limited. PMID- 15133195 TI - Variation in adult life history and stress resistance across five species of Drosophila. AB - Dry weight at eclosion, adult lifespan, lifetime fecundity, lipid and carbohydrate content at eclosion, and starvation and desiccation resistance at eclosion were assayed on a long-term laboratory population of Drosophila melanogaster, and one recently wild-caught population each of four other species of Drosophila, two from the melanogaster and two from the immigrans species group. The relationships among trait means across the five species did not conform to expectations based on correlations among these traits inferred from selection studies on D. melanogaster. In particular, the expected positive relationships between fecundity and size/lipid content, lipid content and starvation resistance, carbohydrate (glycogen) content and desiccation resistance, and the expected negative relationship between lifespan and fecundity were not observed. Most traits were strongly positively correlated between sexes across species, except for fractional lipid content and starvation resistance per microgram lipid. For most traits, there was evidence for significant sexual dimorphism but the degree of dimorphism did not vary across species except in the case of adult lifespan, starvation resistance per microgram lipid, and desiccation resistance per microgram carbohydrate. Overall, D. nasuta nasuta and D. sulfurigaster neonasuta (immigrans group) were heavier at eclosion than the melanogaster group species, and tended to have somewhat higher absolute lipid content and starvation resistance. Yet, these two immigrans group species were shorter-lived and had lower average daily fecundity than the melanogaster group species. The smallest species, D. malerkotliana (melanogaster group), had relatively high daily fecundity, intermediate lifespan and high fractional lipid content, especially in females. D. ananassae (melanogaster group) had the highest absolute and fractional carbohydrate content, but its desiccation resistance per microgram carbohydrate was the lowest among the five species. In terms of overall performance, the laboratory population of D. melanogaster was clearly superior, under laboratory conditions, to the other four species if adult lifespan, lifetime fecundity, average daily fecundity, and absolute starvation and desiccation resistance are considered. This finding is contrary to several recent reports of substantially higher adult lifespan and stress resistance in recently wild-caught flies, relative to flies maintained for a long time in discrete generation laboratory cultures. Possible explanations for these apparent anomalies are discussed in the context of the differing selection pressures likely to be experienced by Drosophila populations in laboratory versus wild environments. PMID- 15133197 TI - Synthesis and biological activity of novel retinamide and retinoate derivatives. AB - Retinoic acid and its amide derivative, N-(4-hydroxyphenyl)retinamide (4-HPR), have been proposed as chemopreventative and chemotherapeutic agents. However, their low cytotoxic activity and water solubility limit their clinical use. In this study, we synthesized novel retinoid derivatives with improved cytotoxicity against cancer cells and increased hygroscopicity. Our syntheses were preceded by selective O-acylation and N-acylation, which led to the production of retinoate and retinamide derivatives, respectively, in one pot directly from aminophenol derivatives and retinoic acid without protection. Transcription assays in COS-1 cells indicated that the N-acylated derivatives (2A-5A) and 4-HPR (1A) were much weaker ligands for all three subtypes of retinoic acid receptor (RAR) than all trans retinoic acid (ATRA), although they showed some selectivity for RARbeta and RARgamma. In contrast, the O-acylated retinoate derivatives (1B-5B) activated all three RAR isotypes without specificity to an extent similar to ATRA. The cytotoxicity was determined using an MTT assay with HCT116 colon cancer cells, and the IC(50) of N-acylated retinamide derivative 4A and O-acylated retinoate derivative 5B was 1.67 microM and 0.65 microM, respectively, which are about five and 13-fold better than that of 4-HPR (8.21 microM), a prototype N-acylated derivative. When retinoate derivative 5B was coupled to organic acid salts, the resulting salt derivatives 5C and 5D had RAR activation and cytotoxicity similar to those of 5B. These data may delineate the relationship between the structure and function of retinoate and retinamide derivatives. PMID- 15133196 TI - Latitudinal clines in Drosophila melanogaster: body size, allozyme frequencies, inversion frequencies, and the insulin-signalling pathway. AB - Many latitudinal clines exist in Drosophila melanogaster: in adult body size, in allele frequency at allozyme loci, and in frequencies of common cosmopolitan inversions. The question is raised whether these latitudinal clines are causally related. This review aims to connect data from two very different fields of study, evolutionary biology and cell biology, in explaining such natural genetic variation in D. melanogaster body size and development time. It is argued that adult body size clines, inversion frequency clines, and clines in allele frequency at loci involved in glycolysis and glycogen storage are part of the same adaptive strategy. Selection pressure is expected to differ at opposite ends of the clines. At high latitudes, selection on D. melanogaster would favour high larval growth rate at low temperatures, and resource storage in adults to survive winter. At low latitudes selection would favour lower larval critical size to survive crowding, and increased male activity leading to high male reproductive success. Studies of the insulin-signalling pathway in D. melanogaster point to the involvement of this pathway in metabolism and adult body size. The genes involved in the insulin-signalling pathway are associated with common cosmopolitan inversions that show latitudinal clines. Each chromosome region connected with a large common cosmopolitan inversion possesses a gene of the insulin transmembrane complex, a gene of the intermediate pathway and a gene of the TOR branch. The hypothesis is presented that temperate D. melanogaster populations have a higher frequency of a 'thrifty' genotype corresponding to high insulin level or high signal level, while tropical populations possess a more 'spendthrift' genotype corresponding to low insulin or low signal level. PMID- 15133198 TI - A novel podophyllotoxin lignan from Justicia heterocarpa. AB - Chromatographic separation of the extract of Justicia heterocarpa T. ANDERS. afforded, in addition to known fatty acids, terpenoids and steroids, a new podophyllotoxin lignan. Structures were elucidated by spectroscopic methods, and the structure of the new lignan was confirmed by single crystal X-ray diffraction studies, which have shown that there is a H-bonding stabilized dimer. PMID- 15133199 TI - Rheological analysis and quantitative evaluation of wet kneaded wax matrix. AB - The properties of the wet kneaded wax matrix were evaluated using a compression tester, whereby a newly proposed sigma index for the plastic deformation was assessed in the pressure transmission diagram. The sigma index was indicative of a characteristic of the plastic yield point in the rheological behavior, and presented an initial and abrupt deformation of wet kneaded mass when the wet kneaded mass was subjected to the pressure. The value of sigma index was confirmed to decrease along with an increase in the plasticity of wet kneaded mass. The wet mass of wax matrix was prepared under various kneading time, and then extruded. The properties of the extruded granules such as pore volume, strength and dissolution were investigated. As a result, it was found that the sigma index decreased with an increase in kneading time. The granules with small value of sigma index showed few porosities, large strength and slow dissolution. It was demonstrated that the sigma index linked the characteristics of wet kneaded mass to the dissolution and the other granule properties. Existence of this link was revealed by sigma index evaluation relevant to the plasticity. The sigma index could be a decisive criterion to permit an in-process evaluation of the kneading progress quantitatively, and also useful for anticipating the dissolution of the final granules roughly. PMID- 15133200 TI - Electronic structure of brain: structure-activity relationships between electronic structure and neurotransmitters based on molecular hardness concept. AB - In order to understand the relation between the electronic structure of neurotransmitters and the brain, a model of the brain based on absolute hardness (eta) and absolute electronegativity (chi) is described. It was found that the coordinate r(chi, eta) of electronic structures of neurotransmitters obtained using the parameters eta and chi can be graphically classified into three groups: catecholamine type (group I), gamma-aminobutanoic acid (GABA) type (group II), and acetylcholine (ACh) type (group III) in the eta-chi diagram. The results suggest that the brainstem and neocortex in the brain are chemically soft and hard, respectively, because they show that the myelinated nerve is chemically soft and the unmyelinated nerve is chemically hard. If one calculates the r(chi, eta) to understand which group a drug belongs to, one can predict the target receptors of the drug from the eta-chi diagram. Using eta-chi maps, one is then able to design medications like antidepressants, tranquilizers, and ACh agonists. PMID- 15133201 TI - Grinding-induced equimolar complex formation between thiourea and ethenzamide. AB - We prepared and characterized a grinding-induced equimolar complex of thiourea with ethenzamide. When thiourea and ethenzamide were co-ground at a molar ratio of 3 : 1, new powder X-ray diffraction (PXRD) peaks were observed in addition to PXRD peaks of thiourea crystals. The optimum stoichiometry of the new structure was confirmed as 1 : 1 mol/mol. Effect of grinding time on the thiourea ethenzamide equimolar complex formation was investigated by using PXRD, differential scanning calorimetry and Fourier transform infrared spectroscopy. The equimolar crystal structure was confirmed by X-ray diffraction measurements of the single crystal which was recrystallized from ethanol. It was found that the intermolecular hydrogen bond formations between thiourea and ethenzamide molecules contributed to the equimolar complex formation. The complex formation was not observed in the cases where benzamide, salicylamide or 3-ethoxybenzamide was co-ground with thiourea. 2-Alcoxyl benzamide structures should be required for the grinding-induced equimolar complex formation with thiourea. PMID- 15133202 TI - Three new norditerpenoid alkaloids from Consolida orientalis. AB - The structures of three new norditerpenoid alkaloids named dehydrodeltatsine (1), 14-O-acetyltakaosamine (2), 18-demethoxypubescenine (3) isolated from the aerial parts of Consolida orientalis (GAY) SCHROD., were elucidated by 1D and 2D NMR spectroscopy and HR-EIMS. Twelve known norditerpenoid alkaloids (type lycoctonine) and the diterpenoid alkaloid ajaconine have been isolated. Several assignments of (13)C-NMR data for delbonine (4) were revised and the complete assignment for 18-hydroxy-14-O-methylgadesine (5) was realized. PMID- 15133203 TI - Novel 17 substituted pregnadiene derivatives as 5 alpha-reductase inhibitors and their binding affinity for the androgen receptor. AB - The in vitro antiandrogenic activity of four new progesterone derivatives: 4, 5, 6 and 7 (8 is a known compound) was determined. These compounds were evaluated as 5alpha-reductase inhibitors as well as by their capacity to bind to the androgen receptor in gonadectomized hamster prostate. The IC(50) value was determined using increasing concentrations of 4, 5, 6, 7 and 8 in the presence of [(3)H]T and the microsomal fraction of the hamster prostate containing the 5alpha reductase enzyme. In this paper we also demonstrated the effect of increasing concentrations of the novel steroids upon [(3)H]DHT binding to the androgen receptors from hamster prostate which produces competition for the androgen receptor sites. The in vitro studies showed that steroids 4, 5, 6, 7 and 8 had an inhibitory activity for the 5alpha-reductase with IC(50) of: 4 (0.17 microM), 5 (0.19 microM), 6 (1 microM), 7 (4.2 microM), and 8 (2.7 microM). On the other hand, the IC(50) value for compounds 4, 5, 6, 7, 8 and DHT showed the following order of affinity for the androgen receptor: 6>7>5>DHT. Surprisingly compounds 4 and 8 did not bind to the androgen receptor. The overall data indicate that all synthesized compounds are inhibitors for the enzyme 5alpha-reductase present in the hamster prostate. In contrast, compounds 5, 6 and 7, which have a cyclohexyl group in the side chain showed a high affinity for the androgen receptor. PMID- 15133205 TI - Investigation of intermolecular interaction in molecular complex of tryptamine and benzoic acid by solid-state 2D NMR. AB - Solid-state NMR spectra and powder X-ray diffraction of the two-component molecular complex composed of tryptamine and benzoic acid were observed to investigate the intermolecular interaction in the molecular complex. 1D (13)C CP/MAS NMR spectrum and powder X-ray diffraction pattern of the complex was clearly different from the convolution of each spectrum of the single component. 2D (1)H-(13)C heteronuclear-correlation (HETCOR) NMR technique indicated that the intermolecular interaction between the primary amine of tryptamine and the carboxyl group of benzoic acid must be related to the complex formation. PMID- 15133204 TI - Synthesis and cytotoxic activity of pyranocarbazole analogues of ellipticine and acronycine. AB - Various 2,2,5,11-tetramethyl- and 2,2,5,6,11-pentamethyl-2,6-dihydropyrano[3,2 b]carbazole derivatives were synthesized by condensation of 3-methylbut-2-enal or 3-chloro-3-methylbut-1-yne with an appropriate hydroxycarbazole. These compounds associate the tricyclic system responsible for the intercalating properties of ellipticine related drugs, with the dimethylpyran pharmacophore of acronycine derivatives. The study of the biological properties of the new pyrano[3,2 b]carbazole derivatives was carried out in vitro on L1210 murine leukaemia cell line. The three (+/-)-cis-diol diesters 15, 16, and 18 were the most active compounds. PMID- 15133206 TI - High performance liquid chromatography with an electrochemical detector in the cathodic mode as a tool for the determination of p-nitrophenol and assay of acid phosphatase in urine samples. AB - Utilizing a commercially available helium-purging device and PEEK tubes for all tubing, especially for connection between the mobile phase and pump, high performance liquid chromatography with an electrochemical detector (ECD/HPLC) at the cathodic mode is a simple and precise method for the determination of p nitrophenol (NP). Studies with cyclic and hydrodynamic voltammetry indicated that 25% aqueous MeOH containing 0.1% (v/v) CF(3)CO(2)H and -0.8 V vs. Ag/AgCl are the best mobile phase and detection potential for cathodic ECD/HPLC. With the present system, the limits of detection and determination were 0.2 and 0.25 microM, respectively, and up to 50 microM, a linear calibration curve was afforded. Within-day precisions for the analysis of 5 and 50 microM NP were 0.8 and 0.7% (n=6), respectively, and between-day precisions (n=6) for these samples were 3.5 and 2.2%, respectively. Compared with the commonly used Bessey-Lowry-Brock method, cathodic ECD/HPLC was useful for the assay of acid phosphatase in urine samples with p-nitrophenyl phosphate disodium salt as a substrate. PMID- 15133207 TI - Terpenoids and aromatic compounds from the New Zealand liverworts Plagiochila, Schistochila, and Heteroscyphus species. AB - A new clerodane- and two new ent-rosane-type diterpenoids have been isolated from the New Zealand liverworts Heteroscyphus billardierii and Plagiochila deltoidea, respectively. The known bisbibenzyl compounds and acetophenones have also been isolated from Schistochila glaucescens and Plagiochila fasciculata. Their structures were established by extensive NMR techniques. Chemosystematics of the Plagiochila species have been discussed. PMID- 15133208 TI - Acid-base cosolvent method for determining aqueous permeability of amiodarone, itraconazole, tamoxifen, terfenadine and other very insoluble molecules. AB - A high-throughput, UV-detection PAMPA (parallel artificial membrane permeability assay) cosolvent procedure is described, based on the use of 20% v/v acetonitrile in aqueous buffer. A training set of 32 drugs (17 bases, 13 acids, 2 ampholytes) was studied both in aqueous buffer and in cosolvent-buffer solutions. A procedure was devised, where intrinsic permeability values, log P(o)(COS), measured in cosolvent solution, are converted to values expected under cosolvent-free conditions, using an in silico model based on Abraham H-bond acidity (alpha) and basicity (beta) descriptors, developed with the Algorithm Builder computer program, to obtain aqueous intrinsic permeability values: log P(o)=0.738+0.885 log P(o)(COS)-1.262alpha+0.436beta, r(2)=0.97, q(2)=0.96, s=0.38, n=32, F=279. Five sparingly-soluble weak bases (solubility <1 microg/ml), which could not be characterized without cosolvent, had their aqueous intrinsic permeability, P(o), estimated: miconazole 0.32 cm/s; itraconazole 3.2 cm/s; amiodarone 13 cm/s; tamoxifen 28 cm/s; terfenadine 162 cm/s. PMID- 15133209 TI - New alpha-tetralonyl glucosides from the fruit of Juglans mandshurica. AB - Five new alpha-tetralonyl glucosides, juglanosides A-E (1-5) were isolated from the fresh rejuvenated fruit of Juglans mandshurica. Their structures were elucidated as (4S)-4-hydroxy-alpha-tetralone 4-O-beta-D-glucopyranoside (1), (4S) 4,5-dihydroxy-alpha-tetralone 4-O-beta-D-glucopyranoside (2), (4S)-4,6-dihydroxy alpha-tetralone 4-O-beta-D-glucopyranoside (3), (4S)-4,5,8-trihydroxy-alpha tetralone 4-O-beta-D-glucopyranoside (4), and (4S)-4,5,8-trihydroxy-alpha tetralone 5-O-beta-D-glucopyranoside (5) on the basis of spectroscopic analysis and chemical evidence. PMID- 15133210 TI - Product analyses of ozone mediated nitration of benzimidazole derivatives with nitrogen dioxide: formation of 1-nitrobenzimidazoles and conversion to benzotriazoles. AB - Several benzimidazole derivatives having electron-withdrawing or -donating substituent(s) at the benzene moiety were used as models of the imidazole moiety of purine bases and their nitration with nitrogen dioxide and ozone (so-called Kyodai nitration) were examined. Products were extracted from the reaction mixture with AcOEt and their structures were analyzed. 1-Nitrobenzimidazole derivatives and unexpected 1-nitrobenzotriazole derivatives were identified. Although the yields of 1-nitrobenzimidazole derivatives were quite low, these were all new compounds that could be obtained only by Kyodai nitration. It was speculated that benzotriazoles were formed via 1-nitrobenzimidazoles and subsequent nitration toward benzotriazoles resulted in the formation of 1 nitrobenzotriazoles. PMID- 15133211 TI - Orally active CCR5 antagonists as anti-HIV-1 agents: synthesis and biological activity of 1-benzothiepine 1,1-dioxide and 1-benzazepine derivatives containing a tertiary amine moiety. AB - The search for orally active CCR5 antagonists was performed by chemical modification of the 1-benzothiepine 1,1-dioxide 3 and 1-benzazepine 4 lead compounds containing a tertiary amine moiety. Replacement of methyl group with a 2-(C(2-4) alkoxy)ethoxy group at the 4-position on the 7-phenyl group of the 1 benzothiepine ring resulted in both enhanced activity and significant improvement in the pharmacokinetic properties upon oral administration in rats. Introduction of C(2-4) alkyl, phenyl or (hetero)arylmethyl groups as the 1-substituent on the 1-benzazepine ring together with the 2-(butoxy)ethoxy group led to further increase of activity. Among the 1-benzazepine derivatives, the isobutyl (6i), benzyl (6o) or 1-methylpyrazol-4-ylmethyl (6s) compounds were found to exhibit highly potent inhibitory effects, equivalent to the injectable CCR5 antagonist 1, in the HIV-1 envelope-mediated membrane fusion assay. In particular, compound 6s showed the most potent CCR5 antagonistic activity (IC(50)=2.7 nM) and inhibitory effect (IC(50)=1.2 nM) on membrane fusion, together with good pharmacokinetic properties in rats. The synthesis of 1-benzothiepine 1,1-dioxide and 1 benzazepine derivatives and their biological activity are described. PMID- 15133212 TI - Glochidiolide, isoglochidiolide, acuminaminoside, and glochidacuminosides A-D from the leaves of Glochidion acuminatum MUELL. AB - The dimeric butenolides glochidiolide and isoglochidiolide, a new glucoside of a nitrogen-containing C(8) dimer, acuminaminoside, and glucosides of C(8) compounds, designated glochidacuminosides A-D, were isolated from the leaves of Glochidion acuminatum. The structures of glochidionolactone and acuminaminoside were determined by X-ray analyses, and those of the remaining C(8) glucosides by NMR spectroscopic analyses, chemical conversion, and a modified Mosher's method. PMID- 15133213 TI - Effect of a new beta-sitosterol analogue on plasma lipid concentrations in rats. AB - N-Substituted succinamic acid beta-sitosteryl ester derivatives were prepared and evaluated. Compounds 1 and 2 were prepared in 76-92% yields by the reaction of beta-sitosterol and succinic anhydride, followed by the activation of the resulting acid compound 1 by thionyl chloride or methyl chloroformate, and finally by amination with appropriate amines. Compound 2a (DANA87) was also easily obtained in one step by the direct addition of beta-sitosterol to dicyclohexylcarbodiimide (DCC) in 80% yield. Administration of the dietary compound DANA87 resulted in significant decreases in total plasma cholesterol (TC) and low-density lipoprotein (LDL) cholesterol concentrations compared with controls in a rat model. High-density lipoprotein cholesterol and plasma triglyceride levels were not affected. These findings indicate that DANA87 functions as TC and LDL cholesterol-reducing agent. PMID- 15133214 TI - Antitrypanosomal activity of Brazilian propolis from Apis mellifera. AB - Extracts from different samples of Brazilian propolis were obtained by Soxhlet extraction or maceration at room temperature using ethanol, water, and accombination of both solvents. Analysis of their composition using HPLC revealed that no major differences were seen when a propolis sample was subject to different extraction methods. The activity of the 15 extracts was assayed against bloodstream trypomastigotes of Trypanosoma cruzi, the etiologic agent of Chagas' disease. Multivariate analysis was applied to evaluate the efficiency of the different extracts and the trypanocidal activity. The extracts could be divided into two groups. In the first, in which, extracts were obtained by reflux in Soxhlet using 100% ethanol, there was a lower content of bioactive compounds and consequently lower trypanocidal activity. Extract 136-Et100 stands out in this group, since it had the highest levels of bioactive compounds together with highest activity against the parasite when compared with all other extracts. The second group comprises extracts with intermediate levels of bioactive compounds and higher activity against T. cruzi. PMID- 15133215 TI - Properties of calcium-induced gel beads prepared with alginate and hydrolysates. AB - Calcium-induced alginate gel beads (Alg-Ca) containing alginate hydrolysate, such as the guluronic acid block (GB), was prepared and the drug release profiles were investigated under simulated gastrointestinal conditions. The addition of GB to Alg-Ca altered its rheological properties. A model drug (hydrocortisone) was incorporated at 78% of its theoretical yield within the dried Alg-Ca containing 5% GB and it was gradually released from the beads in JP XIV 1st medium for disintegration test (pH 1.2), while it was rapidly released with disintegration of the gel matrix in JP XIV 2nd medium (pH 6.8). In contrast, for Alg-Ca containing GB and chitosan, disintegration was not observed in these media and the drug release rate was markedly different. These results demonstrate that the release profiles of drugs incorporated into Alg-Ca can be controlled by adding these polysaccharides. PMID- 15133216 TI - New lathyrane and podocarpane diterpenoids from Jatropha curcas. AB - Chemical investigation on Jatropha curcas resulted in the isolation of twenty constituents among which four diterpenoids were unknown and six compounds, tetradecyl-(E)-ferulate, 3-O-(Z)-coumaroyl oleanolic acid, heudelotinone, epi isojatrogrossidione, 2alpha-hydroxy-epi-isojatrogrossidione, and 2 methyanthraquinone had not been reported earlier from this species. The structures of the new compounds were established by extensive studies of their 1D and 2D-NMR spectra. PMID- 15133217 TI - Phenylethanoid and iridoid glycosides from the Thai medicinal plant, Barleria strigosa. AB - A phenylethanoid (4-hydroxyphenylethyl 4-O-beta-D-glucopyranosyl-(1-->3)-O-alpha L-rhamnopyranoside) and an iridoid (10-O-trans-coumaroyl-eranthemoside) were isolated from an entire Barleria strigosa plant together with verbascoside, isoverbascoside, decaffeoylverbascoside, (+)-lyoniresinol 3alpha-O-beta-D glucoside, apigenin 7-O-alpha-L-rhamnosyl-(1-->6)-O-beta-D-glucoside, 7-O-acetyl 8-epi-loganic acid and (3R)-1-octen-3-ol-3-O-beta-D-xylosyl-(1-->6)-beta-D glucoside. The structural elucidations were based on analyses of physical and spectroscopic data. PMID- 15133218 TI - Antioxidative phenylethanoid and phenolic glycosides from Picrorhiza scrophulariiflora. AB - One new phenylenthanoid glycoside, scroside D (2), was isolated from the roots of Picrorhiza scrophulariiflora (Scrophulariaceae), together with nine known phenylethanoid and phenolic glycosides: 2-(3,4-dihydroxyphenyl)-ethyl-O-beta-D glucopyranoside (1), 2-(3-hydroxy-4-methoxyphenyl)-ethyl-O-beta-D-glucopyranosyl (1-->3)-beta-D-glucopyranoside (3), scroside B (4), hemiphroside A (5), plantainoside D (6), scroside A (7), androsin (8), piceoside (9), and 6-O feruloyl-beta-D-glucopyranoside (10). The structures of these compounds were elucidated using spectroscopic methods. The antioxidative activities of these isolated compounds were evaluated based on their scavenging effects on hydroxyl radicals and superoxide anion radicals, respectively. Compounds 1, 2, and 6 showed potent antioxidative effects as those of ascorbic acid and the structure activity relationship is discussed. PMID- 15133219 TI - Studies on the constituents of Catalpa species. IX. Iridoids from the fallen leaves of Catalpa ovata G. Don. AB - Two new iridoids, 6-O-trans-p-coumaroyl-7-deoxyrehmaglutin A (1) and 6-O-cis-p coumaroyl-7-deoxyrehmaglutin A (2), were isolated from the fallen leaves of Catalpa ovata G. DON. together with six artifact iridoids (3-8). Their structures were established by spectral analysis. In addition, the scavenging effects of the principal compounds isolated from this plant on 1,1-diphenyl-2-picrylhydrazyl radical-scavenging activity were examined. PMID- 15133220 TI - Three new cycloartane triterpene glycosides from Souliea vaginata. AB - Three new cycloartane triterpene glycosides, soulieosides A (1), B (2), and C (3), were isolated from the rhizomes of Souliea vaginata, and their structures were elucidated on the basis of extensive NMR experiments and chemical methods. Soulieosides A-C were assigned as 25-O-acetylcimigenol-3-O-beta-D-(2 acetyl)xylopyranoside (1), 24-O-acetyl-isodahurinol-3-O-beta-D-(2 acetyl)xylopyranoside (2) and 20(S),22(R),23(S),24(R)-16beta:23;22:25-diepoxy 3beta,23,24-trihydroxy-9,19-cyclolanostane-3-O-beta-D-(4-acetyl)xylopyranoside (3), respectively. PMID- 15133221 TI - Aerobic oxidation of thiols to disulfides catalyzed by trichlorooxyvanadium. AB - Thiols were converted into disulfide by the aerobic oxidation catalyzed by trichlorooxyvanadium in the presence of molecular sieves 3A. PMID- 15133222 TI - Structural dependence of HPLC separation pattern of anthocyanins from Bilberry (Vaccinium myrtillus L.). AB - An HPLC method using isocratic elution was established for the analysis of fifteen anthocyanins contained in bilberry (Vaccinium myrtillus L.). Separation was attained by using an aqueous solution of 20% methanol containing 0.5% TFA as the mobile phase with a flow rate of 2 ml/min. The detection limit was 0.3 pmol for delphinidin 3-O-beta-D-glucopyranoside, which is a major anthocyanin in bilberry extract. The reproducibility was 0.19-3.85% (S.E.M) for peak area and 0.64-0.77% (S.E.M) for relative mobility normalized by the elution position of the solvent peak. When the relative elution volumes of each anthocyanins were correlated to their corresponding anthocyanin structures, a characteristic pattern was observed. From this pattern, the structures of unknown anthocyanins could be predicted from their elution times. Therefore, the present method is useful for the study of anthocyanins from various biological sources. PMID- 15133223 TI - Diacylated 8-C-glucosylcyanidin 3-glucoside from the flowers of Tricyrtis formosana. AB - A new diacylated 8-C-glucosylanthocyanin was isolated from the purple flowers of Tricyrtis formosana 'Fujimusume' as one of the major anthocyanins along with four known pigments. The structure of this pigment was determined to be 8-C-(6-O-trans sinapoyl)-beta-glucopyranosylcyanidin 3-O-(6-O-malonyl-beta-glucopyranoside) by chemical and spectroscopic methods. In addition, four known pigments, 8-C glucosylcyanidin 3-malonylglucoside, cyanidin 3-glucoside, cyanidin 3-rutinoside and cyanidin 3-malonylglucoside, were identified as the major anthocyanins in the flowers. PMID- 15133224 TI - Structure-activity relationships of 2-aminothiazole derivatives as inducible nitric oxide synthase inhibitor. AB - Nitric oxide synthase (NOS) has been divided into two major sub-enzymes, i.e. inducible NOS (iNOS) and constitutive NOS (cNOS). Although nitric oxide (NO) plays an important role as host defense mediator, excessive production of NO by iNOS has been involved in the pathology of many inflammatory diseases. Recently, we reported that the 2-imino-1,3-oxazolidine (1a) weakly inhibits iNOS and that introduction of an alkyl moiety on the oxazolidine ring of 1a enhances the inhibitory activity and selectivity for iNOS. In our search for better iNOS inhibitors, we focused our efforts on the 2-aminothiazole scaffold 3 as it possesses a ring similar to that of 1a. In this study, we evaluated the inhibitory activity of a series of 2-aminothiazole derivatives against both iNOS and neuronal NOS (nNOS). Our results show that introduction of appropriately sized substituents at the 4- and 5-position of the 2-aminothiazole ring improves the inhibitory activity and selectivity for iNOS. We also found that the selectivity of 5a [5-(1-methyl)ethyl-4-methylthiazol-2-ylamine] and 5b [5-(1,1 dimethyl)ethyl-4-methylthiazol-2-ylamine] for iNOS was similar to that of oxazolidine derivative 1b (4-methyl-5-propyl-2-imino-1,3-oxazolidine) and much higher than that of L-NAME. However, we could not enhance the inhibitory activity against iNOS by introducing an alkyl substituent into the 2-aminothiazole ring as we could in the case of oxazolidine one. On the other hand, introduction of bulky or hydrophilic substituent at any position of the 2-aminothiazole ring remarkably decreased or even abolished the inhibitory activity against NOS. PMID- 15133225 TI - New lignan glucosides from the stems of Tinospora sinensis. AB - Two new lignan glucosides, tinosposides A and B (1 and 2), were isolated from the stems of Tinospora sinensis collected in Hainan Island, China. Their structures were elucidated on the basis of spectroscopic analysis and chemical evidence. PMID- 15133226 TI - Effect of freeze-thawing and polyethylene glycol (PEG) lipid on fusion and fission of phospholipid vesicles. AB - The effect of freeze-thawing on the size of egg yolk phosphatidylcholine (EggPC) vesicles in the presence of 0-40 mol% distearoylphosphatidylethanolamine polyethylene glycol 2000 (DSPE-PEG) was studied. Mean diameters of the vesicles fell into a range of 80-150 nm after 10 times freeze-thawing in spite of their original size. In the process of freeze-thawing, two opponent events, one is fission and the other is fusion, occurred at the same time. DSPE-PEG accelerated the fusion event. PMID- 15133227 TI - Evaluation of homology modeling of the severe acute respiratory syndrome (SARS) coronavirus main protease for structure based drug design. AB - To accelerate the development of drugs against severe acute respiratory syndrome (SARS), we constructed a homology model of the SARS coronavirus main protease using our modeling software, FAMS Ligand&Complex, and released it before the X ray structure was solved. The X-ray structure showed our model as accurately predicted and useful for structure based drug design. PMID- 15133228 TI - Asymmetric Michael reaction promoted by new chiral phase-transfer catalysts. AB - A catalytic asymmetric Michael reaction promoted by new chiral quaternary ammonium salts is described. The products are obtained with moderate ee (up to 75% ee), and the enantioselectivity is strongly dependent on both the substituents on the aromatic rings and the ammonium moiety in the catalysts. PMID- 15133229 TI - Cancer anti-angiogenic therapy. AB - Tumor angiogenesis affords new targets for cancer therapy, since inhibition of angiogenesis suppresses tumor growth by cutting out the supply of oxygen and nutrients. Anti-angiogenic therapy is thought to be free of the severe side effects that are usually seen with cytotoxic anticancer drugs. Furthermore, anti angiogenic therapy is thought not only to eradicate primary tumor tissues, but also to suppress tumor metastases. However, it is uncertain whether this therapy causes tumor regression because it inhibits only angiogenic events. Recently, a novel anti-angiogenic therapy called anti-neovascular therapy (ANET) has become notable. This therapy inflicts indirect lethal damage on tumor cells by damaging newly formed blood vessels using anti-cancer drugs targeting the angiogenic vasculature, since cytotoxic anti-cancer drugs cause damage to proliferating neovascular endothelial cells as well as tumor cells. Moreover, neovascular endothelial cells would not be expected to acquire drug-resistance. Traditional chemotherapy, which directly targets tumor cells, has potential problems such as low specificity and severe side effects. On the contrary, in ANET, severe side effects may be suppressed, since traditional anti-cancer agents are delivered to the neovessels by DDS technology. Besides the usage of DDS technology, anti neovascular scheduling of chemotherapy, or metronomic-dosing chemotherapy, has also been attempted in which anti-cancer drugs are administered on a schedule to damage neovessels. In this review, we describe traditional anti-angiogenic therapy and ANET. We also discuss anti-angiogenic cancer photodynamic therapy (PDT), since PDT is clinically applied to treat age-related macular degeneration (AMD), in which uncontrolled angiogenesis occurs. PMID- 15133230 TI - A tale of early response genes. AB - Immediate early genes (IEG) are rapidly but transiently induced directly by intracellular signaling cascades to alter patterns of gene expression. It has been proposed that histone modifications could be the key to the quick alteration of chromatin structure, as this spread occurs too rapidly to be the consequence of passage of RNA polymerase II. In this review, we will discuss the different modifications on histones and the chromatin remodeling enzymes, allowing the promoter regions of two IEGs, c-fos and c-jun, to be accessed. PMID- 15133231 TI - In vitro and in vivo effects of dantrolene on carbonic anhydrase enzyme activities. AB - The effects of dantrolene were investigated on carbonic anhydrase (CA) enzyme activities in in vitro human and in in vivo Sprague-Dawley rat erythrocytes. For in vitro study, human carbonic anhydrase-I (HCA-I) and -II (HCA-II) were purified by Sepharose 4B-L-tyrosine-sulfanylamide affinity chromatography, rats were used for in vivo study. In vivo and in vitro CA enzyme activity was determined colorimetrically using the CO(2)-hydration method of Wilbur and Anderson. Dantrolene (1.64 x 10(-5)-6.56 x 10(-5) M) showed in vitro inhibitory effects on HCA-I and HCA-II hydratase activity, when determined using the CO(2)-hydratase method. 50% inhibitory concentration (IC(50)) was 4.09 x 10(-5) M for HCA-I and 3.24 x 10(-5) M for HCA-II. Rat erythrocyte CA activity was significantly inhibited by 10 mg/kg dantrolene for up to 3 h (p<0.001) following intraperitoneal administration. In conclusion, Dantrolene inhibited the carbonic anhydrase enzyme activity under in vitro and in vivo conditions. PMID- 15133232 TI - Inhibition of nitric oxide and tumor necrosis factor-alpha (TNF-alpha) production by propenone compound through blockade of nuclear factor (NF)-kappa B activation in cultured murine macrophages. AB - Lipopolysaccharide (LPS)-stimulated macrophages produce large amounts of nitric oxide (NO) by inducible nitric oxide synthase (iNOS). This is an important mechanism in macrophage-induced septic shock and inflammation. In the present study, we tested a synthetic propenone compound, 1-furan-2-yl-3-pyridin-2-yl propenone (FPP-3) for its ability to inhibit the production of tumor necrosis factor-alpha (TNF-alpha) and an inducible enzyme, iNOS, in the LPS-stimulated murine macrophage-like cell line, RAW264.7. FPP-3 consistently inhibited nitric oxide (NO) and TNF-alpha production in a dose dependent manner, with IC(50) values of 10.0 and 13.1 microM, respectively. Western blotting probed with specific anti-iNOS antibodies showed that the decrease in quantity of the NO product was accompanied by a decrease in the iNOS protein level. In cells transiently transfected with nuclear factor (NF)-kappaB promoter-luciferase reporter construct, this compound clearly inhibited the LPS-stimulated NF-kappaB activation. Moreover, this compound inhibited IkappaB-alpha degradation in a concentration and time-dependent manner. These results indicate that FPP-3 inhibits NO production via inhibition of degradation of IkappaB-alpha through NF kappaB activation. PMID- 15133233 TI - Induction of transition and transversion mutations during random mutagenesis PCR by the addition of 2-hydroxy-dATP. AB - A 2-substituted purine nucleotide analog, 2-hydroxy-2'-deoxyadenosine 5' triphosphate (2-OH-dATP), was added to a PCR mixture, to examine its mutagenic potential. The 2-OH-dATP enhanced the total mutation frequency. Interestingly, 2 OH-dATP induced both transition and transversion mutations, including A:T-->G:C, A:T-->C:G and G:C-->T:A mutations. In contrast, other 2-substituted purine nucleotide analogs, 2-aminopurine-2'-deoxyriboside 5'-triphosphate and 2-amino-2' deoxyadenosine 5'-triphosphate, did not affect the total mutation frequency. These results suggest that 2-OH-dATP is useful in random PCR mutagenesis for the in vitro evolution of nucleic acids and proteins, and for analyses of residues in these biomolecules. PMID- 15133234 TI - N-myc downregulated gene 1 is a phosphorylated protein in mast cells. AB - We previously demonstrated that the in vitro maturation of mouse immature bone marrow-derived mast cells into a mature connective tissue mast cell-like phenotype is accompanied by a marked induction of N-myc downregulated gene (NDRG) 1, a cytosolic protein with unknown function. Here we show that NDRG1 undergoes phosphorylation in mast cells. Recombinant NDRG1 was phosphorylated by calmodulin kinase-II, protein kinase (PK) A and PKC in vitro. Deletion of the C-terminal tandem repeats of NDRG1 resulted in increased phosphorylation by PKA and PKC, but not by calmodulin kinase-II. Furthermore, NDRG1 was phosphorylated on serine and threonine residues in mast cells, a process that was accelerated transiently following cell activation. Pharmacologic studies using kinase-specific inhibitors demonstrated that this NDRG1 phosphorylation in mast cells depended on calmodulin kinase-II and PKA, but not PKC. Collectively, our results indicate that NDRG1 is a multiphosphorylated protein in mast cells, and that the kinetics of increased NDRG1 phosphorylation parallels signaling events leading to exocytosis. PMID- 15133235 TI - Association of N-myc downregulated gene 1 with heat-shock cognate protein 70 in mast cells. AB - N-Myc downregulated gene (NDRG) 1 is markedly induced during in vitro maturation of mouse immature bone marrow-derived mast cells (BMMCs) into a mature connective tissue mast cell (CTMC)-like phenotype. However, cellular function of this unique cytosolic protein is currently obscure. In this study, we sought potential NDRG1 binding proteins using yeast two-hybrid analysis and found that NDRG1 is capable of binding to heat-shock cognate protein 70 (Hsc70) both in vitro and in mast cells. The expression of Hsc70 was markedly elevated during the in vitro maturation of BMMCs into CTMC-like cells in accordance with the increased expression of NDRG1. Deletion of the C-terminal hydrophilic tandem repeats from NDRG1 facilitated the interaction with Hsc70 in vitro. Interaction between NDRG1 and Hsc70 was constitutive in mast cells and was not altered following cell activation. Although NDRG1 undergoes phosphorylation (accompanying paper), the binding of NDRG1 to Hsc70 was not affected by this event. Interestingly, the NDRG1-Hsc70 complex transiently appeared in the nuclear fraction of activated mast cells. PMID- 15133236 TI - Feitai attenuates bleomycin-induced pulmonary fibrosis in rats. AB - Pulmonary fibrosis is a common consequence of numerous pulmonary diseases. The current therapeutic approaches for this condition are unsatisfactory. Feitai, a composite formula consisting of several herbs, is used in China as a folk remedy for treating patients with pulmonary tuberculosis. In this study, we extensively investigate the effects and mechanisms of Feitai on bleomycin (BLM)-induced pulmonary fibrosis in rats. One hundred and twenty male Sprague-Dawley rats were randomly divided into four groups, referred to as the saline-water, saline Feitai, BLM-water, and BLM-Feitai groups. Following a single instillation of BLM (5 mg/kg) or saline, rats were orally administered Feitai at a dose of 3 g/kg body weight or sterilized distilled water once daily. Rats were killed at 7, 14, or 28 d post-BLM. Inflammatory cell count, protein concentration, and lactate dehydrogenase activity in bronchoalveolar lavage fluid were measured, and myeloperoxidase activity and lipid peroxide content in lung homogenates were analyzed. Treatment with Feitai inhibited lung fibrotic progression induced by BLM, as indicated by the decrease in lung hydroproline content and lung fibrosis score at 28 d post-BLM. This was accompanied by significant amelioration of BLM induced body weight loss, lung edema, and inflammatory response during the development of lung injury in the acute phase. The results strongly indicate the beneficial effects of Feitai in protecting against BLM-induced pulmonary fibrosis. Furthermore, the inflammatory response and lipid peroxidation were inhibited by Feitai, suggesting that the effect of this formula on BLM-induced lung injury and fibrosis is associated with antiinflammatory and antioxidant properties. PMID- 15133237 TI - General pharmacology of DW-286a, a new fluoronaphthyridone antibiotic: effects on central nervous, cardiovascular, and respiratory systems. AB - DW-286a is a new class of fluoronaphthyridone antibiotic. Its effect on the central nervous system, general behavior, and cardiovascular, respiratory, and other organ systems were studied. The doses given were 30, 50, 100, 150, 300, and 1000 mg/kg and drugs were administered orally. DW-286a did not show any effects on general behavior, motor coordination, analgesic action, seizure and mortality, blood pressure and heart rate, contractile response of isolated guinea pig ileums, and renal function. On the other hand, DW-286a decreased spontaneous locomotor activity from 120 to 240 min after administration at the doses of 300 and 1000 mg/kg and the respiration rate from 30 to 240 min after the administration of doses up to 100 mg/kg. The sleeping time and body temperature were increased significantly in mice at the dose of 1000 mg/kg. DW-286a increased the charcoal transport significantly at doses of 300 and 1000 mg/kg. DW-286a inhibited gastric acid secretion, reduced the volume of the gastric juice and total acidity, and increased the pH dose dependently. Based on the above results, it was concluded that DW-286a affects spontaneous locomotor activity, respiration and body temperature, gastrointestinal transport, gastric secretory action, and hexobarbital sleeping time at high doses. PMID- 15133238 TI - Induction of apoptosis in cultured rat gastric epithelial cells by ipriflavone: comparison with indomethacin. AB - Based on a previous report illustrating severe gastrointestinal side effects with ipriflavone, we examined the effects of ipriflavone on cell death in cultured rat gastric epithelial cells compared with other chemicals, including indomethacin. Low concentrations of ipriflavone, indomethacin, dexamethasone, and estradiol all induced cell death in gastric epithelial cells. DNA from cultured cells treated with ipriflavone showed fragmentation by electrophoresis. Also, some of the cultured cells treated with ipriflavone were positively stained by TUNEL. In order to confirm induction of apoptosis by ipriflavone, rescue from ipriflavone induced cell death with Z-DEVD-FMK (0.02-0.1 mM), which is a caspase 3 inhibitor, or PGE(2) (0.01-10 mM), was tested. Only Z-DEVD-FMK was observed to rescue the cells from cell death. Similar results were obtained with indomethacin, dexamethasone and estradiol. These results suggest that ipriflavone, indomethacin, dexamethasone and estradiol induce cell death of cultured rat gastric epithelial cells by apoptosis. PMID- 15133239 TI - Synthesis, analgesic, anti-inflammatory and antibacterial activities of some novel 2-methylthio-3-substituted quinazolin-4-(3H)-ones. AB - A variety of novel 2-methylthio-3-substituted quinazolin-4-(3H)-ones have been synthesized by reacting (2-methylthio-4-oxo-3H-quinazolin-3-yl)dithiocarbamic acid methyl ester with a variety of amines, the starting material dithiocarbamate was synthesized from methylanthranilate. The title compounds were investigated for analgesic, anti-inflammatory and antibacterial activities. While the test compounds exhibited significant activity, the compounds A1, A2, A3 and A4 shown more potent analgesic activity, and the compound A4 shown more potent anti inflammatory activity than the reference diclofenac sodium. PMID- 15133240 TI - Auxins affected ginsenoside production and growth of hairy roots in Panax hybrid. AB - Hairy roots of interspecific hybrid ginseng (Panax ginseng x P. quinquefolium), induced by Agrobacterium rhizogenes ATCC 15834, grew well in B5 liquid media supplemented with 2.5 microM auxins (3-indole butyric acid (IBA), 1 naphtaleneacetic acid (NAA) and 3-indoleacetic acid (IAA)). The hairy roots cultured in B5 liquid medium supplemented with 2.5 microM IBA showed best growth (6.39 g fresh weight per a flask, at week 8). The highest content of the total ginsenosides was 1.63% as dry weight at week 8 when cultured with 2.5 microM NAA. The different auxins affected the numbers and lateral branching roots. Especially, 2.5 microM IBA promoted the lateral root formation (43.7+/-4.0 roots, at week 8), and 2.5 microM NAA promoted the lateral root growth (45.3+/-5.6 mm, at week 8). The growth and ginsenosides production of 8-week old hairy roots cultured in B5 liquid media supplemented with IBA and NAA combinations were also investigated. Hairy roots produced higher amounts of ginsenosides in B5 liquid media supplemented with 0.5-1.0 microM IBA and NAA combinations than that cultured in B5 liquid media supplemented with only IBA and NAA. The highest yield of ginsenoside was obtained when cultured with 0.5 microM IBA and 1.0 microM IBA combination (6.38 mg per a flask, at week 8). PMID- 15133241 TI - Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and amplification refractory mutation system (ARMS) analyses of medicinally used Rheum species and their application for identification of Rhei Rhizoma. AB - Previously, we have determined marker nucleotides on the chloroplast matK gene to identify Rheum palmatum, R. tanguticum and R. officinale used as Rhei Rhizoma officially. In the present study, we further developed a convenient and efficient identification method on the basis of marker nucleotides with Amplification Refractory Mutation System analysis. On the basis of the nucleotide substitutions at positions 367 and 937 among the three species on the matK gene, at each position two kinds of reverse primers with complementary 3'-terminal nucleotides were designed. Upon PCR amplification using three sets of primers and template DNA from each species, one or two fragments (202 bp or/and 770 bp) were detected. As the resultant three fragment profiles were species-specific, the procedure enabled us to classify the botanic origins of 22 drug samples of Rhei Rhizoma. PMID- 15133242 TI - Pharmacokinetic interactions between Japanese traditional medicine (kampo) and modern medicine (III). Effect of Sho-seiryu-to on the pharmacokinetics of azelastine hydrochloride in rats. AB - Sho-seiryu-to (SST) is widely used herbal formula in Japanese traditional medicine (kampo) to treat allergic diseases. Since Japanese physicians frequently prescribe this formula combined with azelastine hydrochloride, one of anti histamine and anti-allergic medicines, we evaluated the pharmacokinetic interactions between SST and azelastine hydrochloride in rats to obtain the drug information for the prevention from disadvantage or adverse effects by their combined therapy. Oral administration of SST did not influence the plasma concentration profile of azelastine after its intravaneous injection, suggesting that SST would not change the activities of metabolic enzymes such as cytochrome P450s. However, maximum concentration (C(max)) of azelastine after oral administration of azelastine hydrochloride was significantly reduced and mean residence time (MRT) was significantly lengthened when SST was orally administered at 20 times amount of human daily dosage. There was not significant difference in the area under the plasma concentration-time curve (AUC), suggesting that SST might delay the absorption of azelastine without affecting the extent of bioavailability. Since this delay was independent of ephedrine that is a main constituent of SST and that a suppressor for gastric transit, SST might form unsoluble complex with azelastine to reduce its absorption. At the double of human daily dose, SST did not made the absorption of azelastine delay. The possibility that SST reduce the absorption of azelastine hydrochloride could not be denied completely, however, it is suggested that SST would not cause pharmacokinetic interaction with azelastine hydrochloride clinically. PMID- 15133243 TI - Role of polymorphonuclear leukocytes in the resistance of tumor-bearing mice against Candida albicans infection. AB - Tumor-bearing mice showed a significant resistance against Candida albicans intravenous infection. Longer survival was observed in groups of mice inoculated with fungal cells 2-3 weeks after tumor transplantation with allogeneic sarcoma 180, syngeneic methylcholanthrene-induced Meth A fibrosarcoma, and MM 46 mammary carcinoma than in non-tumor-bearing mice inoculated only with fungal cells. This effect was not observed when the mice were infected only 1 week after tumor transplantation. A significant decrease in the number of C. albicans cells in the kidneys was observed in mice inoculated with fungal cells 2-3 weeks after tumor transplantation. In the tumor-bearing mice treated with cyclophosphamide (CY), a remarkable decrease in both the number of peripheral blood polymorphonuclear leukocytes (PMNs) and the defense against challenge with C. albicans cells was observed, as compared with the CY-untreated groups (normal and tumor-bearing mice). A marked increase in the calcium concentration in serum and number, candidacidal activity, active oxygen level, and myeloperoxidase activity of PMNs was observed in the 2-3-week tumor-bearing mice. From these results, it is suggested that PMNs, which accumulated in the 2-3-week tumor-bearing mice, play an important role in the protection from C. albicans infection by increasing the number and the types of killing factors. PMID- 15133244 TI - A citrus flavonoid hesperidin suppresses infection-induced endotoxin shock in mice. AB - Administration of a citrus flavonoid hesperidin (HES) to mice before LPS challenge significantly reduced tumor necrosis factor (TNF)-alpha production in a dose-dependent manner. Treatment of HES 3 h before intraperitoneal (i.p.) infection with 10(8) CFU Salmonella typhimurium aroA resulted in rescue from lethal shock as similar to LPS-nonresponder mice. Not only bacterial numbers in livers and spleens but also plasma LPS levels significantly decreased by pretreatment with HES. In addition, HES markedly suppressed plasma levels of TNF alpha and high mobility group box chromosomal protein 1 (HMGB-1), decreased the number of apoptotic cells in livers and normalized the activated states of blood coagulation factors such as prothrombin time and platelet numbers caused by infection. Pretreatment of LPS with HES suppressed the chromogenic Limulus reaction. PMID- 15133245 TI - Identification of human p450 isoforms involved in the metabolism of the antiallergic drug, oxatomide, and its inhibitory effect on enzyme activity. AB - Oxatomide is an antiallergic drug used for the treatment of diseases mediated by type I allergy. Recently, it has been reported that terfenadine and astemizole, which have antiallergic actions similar to those of oxatomide, show side effects on the cardiovascular system, such as QT prolongation, ventricular arrhythmia and cardiac arrest. This might be because concomitant drugs such as itraconazole inhibit cytochrome P450 3A4 (CYP3A4), the enzyme responsible for degradation of terfenadine and astemizole, and thus the blood concentrations of the drugs are abnormally increased. On the other hand, isoforms of P450 involved in the metabolism of oxatomide have not been clarified. Therefore, we attempted to identify these isoforms using microsome preparations of in vitro expression systems derived from a human lymphoblastoid cell line. Oxatomide was metabolized by CYP2D6-Val and CYP3A4, but not by CYP1A2, CYP2C9-Arg, CYP2C9-Cys or CYP2C19. We also examined whether oxatomide showed inhibitory effects on metabolic activity of individual P450 isozymes using model substrates for each isozyme. Oxatomide did not inhibit the metabolism of the model substrates for CYP1A2, CYP2C9-Arg, CYP2C9-Cys and CYP2C19, but inhibited the degradation of those for CYP2D6-Val and CYP3A4. It was found that oxatomide is metabolized by CYP2D6 and CYP3A4 in human liver microsomes, and simultaneously acts as an inhibitor for these isoforms, responsible for the metabolism of the drug itself. PMID- 15133246 TI - Monoclonal antibodies and sandwich ELISA for quantitation of HM-1 killer toxin. AB - To establish a method for quantitative analysis of HM-1 killer toxin (HM-1), two purified mouse monoclonal antibodies, 1F1 and 4A2, and rabbit polyclonal antiserum against HM-1 were prepared. Both monoclonal antibodies were classified as IgG1(kappa) subtype, and did not neutralize the killing activity of HM-1. By SPOTs analysis, the epitope of 1F1 was found in the sequence of CDPNTG with a corresponding sequence of 11-16 from N-terminal amino acid residues of HM-1, but the epitope of 4A2 was not determined. Using 4A2 and polyclonal antiserum, the sandwich enzyme-linked immunosorbent assay (ELISA) was applied to establish the quantitative determination of HM-1. The concentration of HM-1 was determined successfully at the range of 2.5-100 ng/ml. But in the case of 1F1, the method was not established. Genes were constructed to apply the system to the measurement of the secreted concentrations of mutant HM-1, and it was evident that the production of mutant toxins varied among HM-1 mutant genes. The findings of this study are unique in determinimg the epitope of monoclonal antibody against HM-1, and in quantifying the HM-1 using the spot analysis and sandwich ELISA methods. PMID- 15133247 TI - Effect of Aloe vera leaf gel and pulp extracts on the liver in type-II diabetic rat models. AB - The aim of this work was to investigate the effects of Aloe vera leaf pulp and gel extracts on the liver tissue of neonatal streptozotocin (n0STZ)-induced type II diabetic rats. The diabetic rats were separated into four groups and each group was given the following samples by gavage, daily for 15 d: phosphate buffered saline (PBS; diabetic control), Aloe leaf pulp extract, Aloe leaf gel extract, glibenclamide. Liver tissues were examined histologically. The markers of oxidative stress: glutathione (GSH), non-enzymatic glycosylation (NEG) and lipid peroxidation (LPO), were determined in liver tissue. Biochemical parameters for liver function: serum alkaline phosphatase (ALP), and alanine transaminase (ALP) activities, were evaluated. All parameters were also determined in healthy (non diabetic) rats for comparison. In the diabetic control group, the degenerative changes in liver tissue were remarkable, while in the diabetic groups given Aloe pulp and gel extracts and glibenclamide, the damage to the liver tissue was decreased. The increase of GSH and the decrease of NEG and LPO in liver tissues with the treatment of Aloe gel extract, is consistent with the beneficial effect of Aloe. Serum ALP and ALT activities were also decreased in the groups given Aloe gel extract. It was concluded that Aloe gel extract has a protective effect comparable to glibenclamide against hepatotoxicity produced by diabetes if used in the treatment of type-II diabetes. PMID- 15133248 TI - Girolline, an antitumor compound isolated from a sponge, induces G2/M cell cycle arrest and accumulation of polyubiquitinated p53. AB - Girolline, an antitumor compound isolated from a sponge, has been reported to inhibit the termination step of protein synthesis in vivo. In this study, we found that girolline induced G2/M cell cycle arrest in several tumor cell lines. Immunochemical analysis revealed that polyubiquitinated p53 was accumulated in girolline-treated cells, while other polyubiquitinated cellular proteins were not accumulated, indicating that the effect of girolline is specific for p53. On the other hand, girolline did not inhibit proteasome activity in vitro, and accumulation of polyubiquitinated p53 was scarcely detected in the presence of leptomycin B, an inhibitor of nuclear export. Based on the above findings, we propose that girolline affects the step of recruitment of polyubiquitinated p53 to the proteasome. PMID- 15133249 TI - Antioxidant properties of some medicinal plants: Prangos ferulacea (Apiaceae), Sedum sempervivoides (Crassulaceae), Malva neglecta (Malvaceae), Cruciata taurica (Rubiaceae), Rosa pimpinellifolia (Rosaceae), Galium verum subsp. verum (Rubiaceae), Urtica dioica (Urticaceae). AB - Antioxidant and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activities, reducing powers and the amount of total phenolic compounds of aqueous and/or methanolic extracts of some medicinal plants used in Eastern Turkey were studied. These plants are Prangos ferulacea (CASIR), Sedum sempervivoides (HOROZ LELESI), Malva neglecta (EBEMGUMECI), Cruciata taurica (SARILIK OTU), Rosa pimpinellifolia (KOYUN GOZU), Galium verum subsp. verum (MADAVUR OTU), Urtica dioica (ISIRGAN). The highest peroxidation inhibitions were shown by aqueous extracts of C. taurica and R. pimpinellifolia (IC(50): 0.00022 mg/l and IC(50): 23 mg/l, respectively). However, the highest DPPH radical scavenging activity, reducing power and the amount of phenolic compounds were shown by R. pimpinellifolia. The lowest antioxidant properties were shown by aqueous extract of M. neglecta. PMID- 15133250 TI - Pharmacokinetics and metabolic rates of acetyl salicylic acid and its metabolites in an Otomi ethnic group of Mexico. AB - The objective of this study was to determine pharmacokinetic differences of acetyl salicylic acid (ASA) and its metabolites: gentisic acid (GA), salicylic acid (SA) and salicyluric acid (SUA) between Otomies and Mesticians healthy subjects. Design. Ten Otomies and 10 Mesticians were included. After a single dose of aspirin given orally (15 mg/kg), blood and urine samples were collected at different times. Results. Pharmacokinetic parameters of salicylates showed significant differences, except distribution volume of SA, and elimination half life of SUA. Metabolic rates of ASA showed significant differences for all rates between both groups. On the other hand, percentages of dose excreted were more reduced for SA and SUA for the Otomies than for the Mesticians. Conclusion. Results reflect differences in the hydrolysis way i.e. from ASA to SA and aromatic hydroxylation i.e. from SA to GA, which were slower in Otomies subjects, showing a possible pharmacokinetic differences about the capabilities of ASA biotransformation as a consequence of ethnic differences. PMID- 15133251 TI - Evaluation of in vitro antimicrobial and in vivo cytotoxic properties of some novel titanium-based coordination complexes. AB - The aim of the present study was to determine the antimicrobial and cytotoxic activities of eight novel titanium(III) based coordination complexes [Ti(Pht)(2)(DL-serine)(2), S(1)], [Ti(Pht)(2)(glycine)(2), S(2))], [Ti(Pht)(2)(cystine)(2), S(3)], [Ti(Pht)(2)(DL-leucine)(2), S(4)], [Ti(Suc)(2)(L leucine)(2), S(5)], [Ti(Suc)(2)(cystine)(2), S(6)], [Ti(Suc)(2)(cystein)(2), S(7)] and [Ti(Suc)(2)(DL-serine)(2), S(8)] against several gram-positive and negative bacteria, fungi and brine shrimp nauplii. The investigation showed that almost all of the complexes were moderately active against tested bacteria and fungi at high concentration (200 microg/disc) compared with the standard antibiotic, amoxicillin and the antifungal agent, nystatin. In vivo lethality bioassay experiment showed that only S(7) and S(8) among the complexes had better cytotoxic effect than standard gallic acid. The LC(50) values of these two complexes were found to be 1.00 and 1.21 microg/ml, respectively. Thus the results suggest that only two complexes (S(7), S(8)) among the titanium(III) based coordination complexes show the anticancer properties comparable to the standard cytotoxic agent, and further studies of these two complexes may be helpful for their clinical implication. PMID- 15133252 TI - Inhibitory effect of flavonoids on sulfo- and glucurono-conjugation of acetaminophen in rat cultured hepatocytes and liver subcellular preparations. AB - A large group of flavonoids was investigated for inhibitory effects on sulfo- and glucurono-conjugation of acetaminophen when added to rat cultured hepatocytes and liver subcellular preparations. The flavonoids inhibited the production of both sulfate and glucuronide conjugates in the cultured cells, with potencies that depended on the specific flavonoid. Among the flavonols, quercetin, kaempferol and galangin were much more effective than myricetin and morin. Flavones including luteolin, apigenin and chrysin were as effective as the corresponding three flavonols above. The inhibition of conjugation by other simple flavones such as 3-, 5-, 7- and 3',4'-OH flavones, and by catechins such as epicatechin and epigallocatechin, was very weak. These data suggest that the presence of both C5 and 7 hydroxyl substitutions on the A-ring in the flavone structure is required for effective inhibitory activity. The effect of flavonoids on sulfo- and glucurono-conjugation was also examined by incubating acetaminophen with isolated liver cytosolic and microsomal preparations, respectively. The active flavonoids in the cells remarkably inhibited the sulfation, but not glucuronidation, in cell-free enzymatic preparations in vitro. The mechanism of inhibition of conjugation by flavonoids in cultured hepatocytes is not likely to depend on the direct inhibition of sulfo- and glucurono-transferase activity by flavonoids. PMID- 15133253 TI - Molecular characterization of pharmacological properties and selectivity of SWR 0315NA for beta3-adrenoceptors. AB - The pharmacological properties of SWR-0315NA, (E,Z)-[4[[1-[2-[(3-phenoxy-2 hydroxy propyl)]amino]ethyl]-1-propenyl]phenoxy]acetic acid sodium, were compared with those of (-)-isoproterenol. In the radioligand binding studies of [(125)I]iodocyanopindolol with COS-7 cell membranes that transiently expressed human beta-adrenoceptor (beta-AR) subtypes, SWR-0315NA exhibited 1-fold and 2 fold greater binding affinities for beta(3)-AR than those for beta(1)- and beta(2)-ARs, respectively. The maximal stimulatory effects of SWR-0315NA on cAMP accumulation in CHO cells expressing all the beta-AR subtypes were 79%, 3% and 93% for beta(1)-, beta(2)- and beta(3)-ARs of those produced by (-) isoproterenol, respectively. SWR-0315NA has 26.3-fold and more than 630-fold greater selectivity for beta(3)-AR than those for beta(1)- and beta(2)-ARs in potency, respectively. These results indicate that although SWR-0315NA has lower binding selectivity towards beta-AR subtypes, it is a selective agonist with high intrinsic activity for beta(3)-AR as compared with (-)-isoproterenol. PMID- 15133254 TI - Danthron inhibits the neurotoxicity induced by various compounds causing oxidative damages including beta-amyloid (25-35) in primary cortical cultures. AB - Oxidative stress caused by an elevation in reactive oxygen species (ROS) plays an important role in Alzheimer's disease and other neurodegenerative diseases. In this study, we examined the neuroprotective effect of danthron (1,8 dihydroxyanthraquinone) against neurotoxicities induced by beta-amyloid (25-35), excitotoxins, apoptosis, and oxidative stress in primary cortical cultures. Danthron dose-dependently reduced neuronal injury induced by 30 microM beta amyloid (25-35). Danthron significantly inhibited oxidative injury induced by 100 microM Fe(3+) and decreased membrane lipid peroxidation induced by 100 microM Fe(3+) as measured by thiobarbituric-acid-reactive substance (TBARS). Danthron (0.5-50 microM) ameliorated the effects of buthionine sulfoximine (BSO, 1 mM), which depletes endogenous glutathione by 10-73%. Danthron also dose-dependently inhibited neuronal injury mediated by nitric oxide (NO) radicals, but failed to inhibit injury due to superoxide radicals (O(2-)). These results suggest danthron treatment may, in part, reduce neurotoxicity related to beta-amyloid protein by both dominant inhibitory effects on membrane lipid peroxidation and glutathione deprivation. PMID- 15133255 TI - Inhibitory effect of propolis on the growth of human leukemia U937. AB - We have investigated the effect of propolis (CB Propolis) on the growth of human histiocytic lymphoma U937 cells. We found that propolis strongly inhibited the growth of the cells and macromolecular synthesis in a dose- and time-dependent manner by apoptosis. Propolis at 0.015-0.5 microl/ml showed antitumor activity with an IC(50) of 0.18 microl/ml for 3 d. It also inhibits DNA, RNA and protein synthesis with an IC(50) of 0.08, 0.17 and 4.3 microl/ml, respectively. The inhibitory effect on DNA synthesis was partially irreversible. Moreover, an apoptotic DNA ladder and chromatin condensation were observed in the same concentration range in which cell growth was inhibited. The caspase inhibitor, Z Asp-CH(2)-DCB, prevented DNA fragmentation. These results suggest that the antitumor activity of propolis occurs through the induction of apoptosis. Propolis may be useful as a cancer chemopreventive and chemotherapeutic agent. PMID- 15133256 TI - In vitro propagation by asymbiotic seed germination and 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical scavenging activity studies of tissue culture raised plants of three medicinally important species of dendrobium. AB - A simple and efficient plant propagation system has been developed by asymbiotic germination of seeds in three medicinally important Dendrobium species, namely, Dendrobium tosaense, Dendrobium moniliforme, and Dendrobium linawianum. Plants obtained from natural habitats were grown in the greenhouse. The flowers were hand pollinated. Seeds of the capsules derived after 12 weeks of hand-pollination germinated asymbiotically (50-74%) on half strength Murashige and Skoog's (MS) basal medium with 3% sucrose and solidified with 0.9% Difco agar. Active growth in the germinated seedlings was achieved by re-culturing on full strength MS basal medium supplemented with 8% banana homogenate, 8% potato homogenate, 8% coconut water, 1.5% sucrose and 0.9% Difco agar. Healthy plantlets, transferred to plastic trays containing moss or moss and tree fern, successfully acclimatized (84-100%) in the greenhouse. A marked varied response was observed in the free radical scavenging activity of methanolic extracts of in vitro propagated plants, on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical using a UV spectrophotometer assay. Methanolic extracts were prepared by dissolving the powdered plant material, obtained from six months old in vitro propagated plants, each about 5 g, in boiling methanol. The percentage of scavenging effect of D. tosaense extract was 95.9% at 0.4 mg/ml concentration, whereas D. monoliforme, and D. linawianum extracts scavenged 83.4% and 92.3%, respectively, at a concentration of 0.4 mg/ml. All the extracts scavenged DPPH radical significantly in a concentration dependent manner. PMID- 15133257 TI - Reducing activity of colloidal platinum nanoparticles for hydrogen peroxide, 2,2 diphenyl-1-picrylhydrazyl radical and 2,6-dichlorophenol indophenol. AB - Shimizu and Tsuji established a method of preparing colloidal platinum nanoparticles, whose average size is 2 nm, by ethanol reduction of H(2)PtCl(6) in the absence of protective agents for the particles. Platinum nanoparticles have negative surface potential and are stably suspended from an electric repulsion between them. The platinum nanoparticles reduced hydrogen peroxide and 2,2 diphenyl-1-picrylhydrazyl radical (DPPH) below 0.1 ppm. It is necessary to use higher concentration of platinum nanoparticles for the reduction of 2,6 dichlorophenol indophenol (DCIP) than that of hydrogen peroxide and 2,2-diphenyl 1-picrylhydrazyl radical, because reoxidation of DCIPH(2) (reduced) by oxygen was not negligible under our experimental conditions. These results indicate that electrons on platinum nanoparticles produced by the method of Shimizu and Tsuji can reduce hydrogen peroxide, DPPH radical or DCIP transferring electrons. PMID- 15133258 TI - So-Cheong-Ryong-Tang, tradititional Korean medicine, suppresses Th2 lineage development. AB - The present study was designed to evaluate the effect of So-Cheong-Ryong-Tang (SCRT, also called Sho-Seiryu-To or Xiao-Qing-Long-Tang) on helper T cell development by monitoring Th1/Th2-specific cytokine secretion patterns in artificially induced Th1 or Th2 polarized conditions. The results demonstrated that Th2 cells were dramatically underpopulated in the Th2-driven condition triggered by SCRT treatment, while the Th1 cells were not altered in the Th1 skewed condition. Furthermore, under Th2-skewed conditions the levels of interleukin-4 were considerably decreased with SCRT treatment. The expression of GATA-3, a transcription factor that plays a pivotal role in Th2 lineage programming, did not change with SCRT treatment, while the expression of another Th2 transcription factor, c-Maf, was dramatically suppressed. These data suggest that SCRT modulates Th2 development by suppressing c-Maf expression. This study implies that the SCRT effect on CD4(+) T cells is a key pharmacologic point of effect for treating IgE-mediated allergic asthma. These results also suggest that SCRT might be a useful agent for the correction of Th2-dominant pathologic disorders. PMID- 15133259 TI - Structure-activity relationship of oleanane disaccharides isolated from Akebia quinata versus cytotoxicity against cancer cells and NO inhibition. AB - In order to further determine the nature of structure-activity relationship on the cytotoxicities of saponins with 1-->2 and 1-->3 linkages of disaccharides, we isolated guaianin N, collinsonidin, kalopanaxsaponin A and hederoside D(2) as disaccharides, and patrinia glycoside B-II as a trisaccharide, from the n-BuOH extract of Akebia quinata (Lardizabalaceae). Complete acid hydrolysis of the extract afforded oleanolic acid (1) and hederagenin (2). By sulforhodamine B (SRB) assay, kalopanaxsaponin A containing an alpha-L-rhap-(1-->2)-alpha-L-arap moiety exhibited distinctly higher cytotoxicity (IC(50) 1.8-2.7 microg/ml) against all of the tested cell lines than the other saponins (IC(50), 4-8 microg/ml). These results suggest that the alpha-L-rhap-(1-->2)-alpha-L-arap moiety has a unique structural significance in terms of its cell biochemistry, compared to those oleanane glycosides with other sugar linkages. On the other hand, kalopanaxsaponin A exhibited a significant inhibitory effect on nitric oxide production by lipopolysaccharide (LPS)-activated macrophage 264.7, whereas other saponins had weaker activities. PMID- 15133260 TI - Antimalarial activity of lavandulyl flavanones isolated from the roots of Sophora flavescens. AB - Four lavandulyl flavanones, (2S)-2'-methoxykurarinone (1), sophoraflavanone G (2), leachianone A (3), and (-)-kurarinone (4), which are isolated from the roots of Sophora flavescens have been tested for in vitro antimalarial activity against Plasmodium falciparum. Compounds 1-3 showed moderate antimalarial activities with EC(50) values of 2.4 x 10(-6), 2.6 x 10(-6), and 2.1 x 10(-6) M, respectively. These compounds did not show selective toxicity against P. falciparum in the toxicity test on mouse mammalian tumor cells, however, it is suggested that the position of methoxyl groups in flavanone skeleton plays an important role on antimalarial activity. PMID- 15133261 TI - Farnesol, a morphogenetic autoregulatory substance in the dimorphic fungus Candida albicans, inhibits hyphae growth through suppression of a mitogen activated protein kinase cascade. AB - Candida albicans grew in hyphal form in RPMI1640, however, addition of farnesol inhibited the formation. Farnesol did not affect the expression of mRNAs related to cAMP-EFG1 pathways. The mRNAs (HST7 and CPH1) in mitogen activated protein kinase (MAP) cascades were decreased in farnesol-treated cells, but CST20 was not. Furthermore, expression of general amino acid permease 1 (GAP1) was decreased by farnesol. We concluded that farnesol inhibits a MAP kinase cascades, and the suppression is a cause of interruption of hyphae formation. PMID- 15133262 TI - Increase of lactoferrin concentration in mastitic goat milk. AB - Lactoferrin concentration (LFC) in normal and mastitic milk of dairy goats were examined. LFC in bulk milk collected from 70 dairy goat farms and individual milk samples from 10 goats with mastitis were measured by enzyme-linked immunosorbent assay (ELISA) and their reaction time in methylene blue reduction test (MBRT) monitored. Bulk milk samples were categorized into three grades, such as high, normal and low, based on the reaction time in MBRT. The mean LFC in milk that was considered high quality (167 microg/ml) was significantly lower than that of those graded as normal (218 microg/ml) and low quality (304 microg/ml), while mean LFC in mastitic milk was 587 microg/m l. The correlation coefficient between milk LFC and MBRT time was found to be -0.7. Three goats were inoculated with Staphylococcus aureus into one of their udder halves. The mean milk LFC was found to be significantly higher (1500 microg/ml) than the control (30 microg/ml). These findings suggest that milk LFC may be useful as an index for intramammary infection in goats. PMID- 15133263 TI - Expression of constitutive endothelial, neuronal and inducible nitric oxide synthase in the testis and epididymis of horse. AB - The expression of three isoforms of nitric oxide synthase (NOS) were examined in the testis and epididymis of a thoroughbred horse. Immunohistochemical studies demonstrated the presence of eNOS immunostaining in some germ cells in the seminiferous tubules and in vascular endothelial cells in the interstitial tissues. Interstitial cells, most likely Leydig cells, were also intensely immunopositive for eNOS. The pattern of immunostaining for nNOS was similar to that for eNOS in the testis. Weak expression of iNOS was detected in the seminiferous tubules of the testis, but intense expression was found in interstitial cells. Inducible NOS was also strongly detected in stereocilia, sperm, epithelium and connective tissue of the epididymis of normal horses. These findings suggest that three isoforms of NOS are expressed in the testis and epididymis of horse and that they play important roles in the biology of interstitial cells that produce testosterone, as well as in spermatogenesis in the seminiferous tubules. PMID- 15133264 TI - Suppression of cytokine-inducible nitric oxide synthesis during intraperitoneal meth a tumor growth. AB - Nitric oxide (NO*) synthesis is induced within many tumors. The timecourse of NO* synthesis was evaluated during intraperitoneal Meth A fibrosarcoma progression. While increasing macrophage recruitment into ascites was noted, inducible nitric oxide synthase (iNOS) antigen and function peaked between days 3-6 after tumor implantation. The capacity of cells to respond to LPS and IFNgamma stimulation was markedly depressed on day 9 and 11. Cellular proliferation correlated in an inverse fashion with levels of NO* synthesis. Electron paramagnetic resonance spectroscopy and nitrotyrosine immunostaining failed to show accumulation of characteristic target cell lesions induced by NO*. These findings lead us to conclude that NO* production was increasingly suppressed during Meth A tumor progression. Depression of NO* production did not correlate with levels of the inhibitory cytokines TGFbeta and IL-10, but could be partially overcome by addition of sepiapterin (a tetrahydrobiopterin prodrug). Thus, depletion of essential co-factors necessary for iNOS function may contribute to depressed NO* responses during cancer progression. PMID- 15133265 TI - Development of reverse transcriptase PCR and nested PCR to detect porcine hemagglutinating encephalomyelitis virus. AB - Porcine hemagglutinating encephalomyelitis virus (HEV) causes encephalomyelitis, or vomiting and wasting disease, in suckling piglets. The mortality rate for piglets under 3 weeks old is 100%, but they are usually protected by maternal antibodies. Recently, the risk of an HEV outbreak has increased in the pig industry, because of widely using specific pathogen-free pigs that have no antibodies to HEV. We developed reverse transcription (RT) PCR and nested PCR to detect HEV. Primer sets of polymerase, non-structural protein, and spike protein were designed for RT-PCR and nested PCR based on the nucleotide sequences of the HEV 67N strain. The PCR designated primer sets of spike protein detected only HEV viral RNA among other related nidoviruses. Detection of HEV viral RNA by nested PCR was more sensitive than virus isolation in cell cultures. Nested PCR detected HEV viral RNA from experimentally infected samples of mice and field samples of piglets. The RT-PCR and nested PCR methods to detect HEV is considered a good way to show the HEV etiology on pig farms. PMID- 15133266 TI - Quantitative study of pulmonary endocrine cells in fetal, postnatal and adult sheep. AB - Using light microscopic immunohistochemistry, neuron-specific enolase (NSE) positive endocrine cells were quantitatively analyzed in the sheep lung during different stages of development from the canalicular stages to adulthood. In all stages, NSE-positive endocrine cells were usually located in the bronchi and bronchioles as solitary cells, although a few NSE-positive cell clusters, the so called neuroepithelial bodies, were found in some places. The number of NSE positive endocrine cells decreased with advanced stages of gestation. In the late alveolar stage, the number of NSE-positive endocrine cells reached its bottom during the fetal period. There was a gradual upturn after birth. The overall pattern of growth and differentiation of the endocrine cells is most likely species-related and depends on the state of airway development; the number of the endocrine cells of almost all animals, excluding the sheep, in relation to the size of the lung reaches a peak in the late fetal and early neonatal periods and decreases shortly thereafter. NSE-positive endocrine cells were also predominantly located in the large airways during the early stage of development (canalicular stage), and were found more frequently in the small peripheral airways towards the term. These results show the number of NSE-positive endocrine cells in the sheep to be different from that seen in other species. PMID- 15133267 TI - Transport stress increases somatic cell counts in milk, and enhances the migration capacity of peripheral blood neutrophils of dairy cows. AB - The present study was designed to determine the effects of physiological stress on milk-somatic cell counts (SCC) and function of bovine peripheral blood leukocytes (PBL). Nine healthy lactating cows were used in the examination. Five cows were transported 100 km for 4 hr (transported group; TG), and 4 cows were penned (non-transported group; NTG). Blood and milk samples were collected at 0, 2, and 4 hr after loading, and at 2 hr, and 1, 2, 3, and 6 days after unloading. The following activities were measured: adhesion receptor (CD 18 and L-selectin) expression of neutrophils and monocytes, migration capacity and percentage of apoptotic cells of neutrophils, serum soluble L-selectin (sL-selectin), plasma cortisol, and SCC. A significant increase in plasma cortisol and milk SCC was observed in TG. Leukocytosis, derived from neutrophils was recorded in TG, and was indicated by apoptotic measurement as an increase of young cells from the marginal pool. Increased migration and decreased surface expression of both L selectin and CD 18 in neutrophils were observed after transportation. Elevated serum sL-selectin was also noted as a result of transportation. The present study indicated that transport stress modulates peripheral blood neutrophil function, particularly enhancing migration capacity, and causes diapedesis across the mammary epithelium. Increased milk SCC in transported cattle might be due to these phenomena, and severe physiological stress may bring about an increase in SCC in milk. PMID- 15133268 TI - Inhibitory effect of pyrimidine and purine nucleotides on the multiplication of Babesia gibsoni: possible cause of low parasitemia and simultaneous reticulocytosis in canine babesiosis. AB - The present study was conducted to determine the cause of low parasitemia and simultaneous reticulocytosis in canine babesiosis. The parasitemia was significantly decreased in in vitro cultures of Babesia gibsoni by the pretreatment of host canine erythrocytes with lead acetate, which is a specific inhibitor of pyrimidine 5'-nucleotidase subclass I (P5N-I). The serum from dogs chronically infected with B. gibsoni did not decrease the activities of hexokinase, glucose-6-phosphate dehydrogenase or 6-phosphogluconate dehydrogenase in canine reticulocytes, although it was previously reported that this serum had inhibitory effects on both the maturation of reticulocytes and the canine P5N-I and purine-specific 5'-nucleotidase activities. Furthermore, the in vitro multiplication of B. gibsoni was significantly inhibited by pyrimidine nucleotides such as cytidine 5'-monophosphate (5'-CMP), which is preferentially catalyzed by P5N-I and also inhibits the morphological maturation of canine reticulocytes. Purine nucleotides such as inosine 5'-monophosphate (5'-IMP) also had an inhibitory effect on the multiplication of this parasite. These results suggest that nucleotides such as 5'-CMP and 5'-IMP might accumulate in young erythrocytes and/or serum in dogs infected with B. gibsoni as a result of the decreased activity of erythrocyte 5'-nucleotidase, and the accumulation of these nucleotides might inhibit the multiplication of this parasite and simultaneously retard the maturation of reticulocytes. The results obtained from the in vitro examinations in the present study may partially clarify the relationship between low parasitemia and simultaneous reticulocytosis in vivo in canine babesiosis. PMID- 15133269 TI - Histopathological findings of cleft palate in rat embryos induced by triamcinolone acetonide. AB - Triamcinolone acetonide (TAC), a synthetic glucocorticoid, induces cleft palate resulting from poor development of palatal shelves in mice. However, TAC has no effect on medial edge epithelial cells (MEE cells) in secondary palatal shelves. In the present study, we examined the relationship between the pathogenesis of cleft palate and the effects on MEE cells and palatal mesenchymal cells in rat embryos/fetuses exposed to TAC. Pregnant Wistar Hannover rats were given TAC intramuscularly at 0.5 mg/kg at gestation days (Day) 12, 13, and 14, then embryos/fetuses were harvested on Days 14.5, 15, 16 and 20. The effects of TAC were as follows; an inhibition of palatal mesenchymal cell proliferation on Day 14.5, a decrease in the density of palatal mesenchymal cells and MEE cells, and expression of epidermal growth factor (EGF) receptors in MEE cells on Day 15, and stratified squamous differentiation of MEE cells with expression of cytokeratin and EGF receptors on Day 16. These findings indicated that TAC inhibited the proliferation of mesenchymal cells and affected the differentiation of MEE cells into stratified squamous epithelia in the palatal shelves of rat embryos. However, these stratified squamous MEE cells partially fused with each other. Thus, we suspected that a major contributing factor to the formation of TAC induced cleft palate might not be the altered differentiation of MEE cells, but the inhibition of mesenchymal cell proliferation. PMID- 15133270 TI - Characterization of stage-specific and cross-reactive antigens from Eimeria acervulina by chicken monoclonal antibodies. AB - The characterization of five chicken monoclonal antibodies (mAbs) that were developed against apical complex antigens of Eimeria acervulina sporozoites is realized and the mAbs reactivity to merozoites belonging to this species is tested. Using immuno-fluorescence assay (IFA), one mAb (HE-4) that recognized apical antigens common to sporozoites of E. acervulina and E. brunetti bound antigens localized on the apical tip of merozoites from all stages of development examined. The mAb 8E-1, reactive with antigens found on the apical tip of all chicken Eimeria sporozoites, also showed binding to antigens common to merozoites from all generations. Another mAb, 8C-3, which identified an antigen shared by sporozoites apical tip and sporocysts wall of E. acervulina reacted very weak and inconstantly with the merozoites from all generations whereas the mAbs 5D-11 and 8D-2 that recognized antigens shared by the sporozoites of E. acervulina and E. maxima (mAb 5D-11) and E. acervulina and E. brunetti (mAb 8D-2) did not react with the merozoites from any generation. Collectively, these results showed that the invasive stages of chicken Eimeria share cross reactive apical complex antigens which are inter-species and inter-generation-specific that might be components of a potential recombinant vaccine. PMID- 15133271 TI - Role of actin microfilaments in canine distemper virus replication in vero cells. AB - Several studies have indicated that viruses require a specific cytoskeletal structure for replication in host cells. In this study, we examined the role of actin fiber in the replication of canine distemper virus (CDV), belonging to the Morbillivirus genus of the family Paramyxoviridae. For this purpose, we used two actin depolymerizing agents, cytochalasin-D (C-D) and mycalolide-B (ML-B). In Vero cells, C-D disrupted actin fibers distributed in the cytosol, but peripheral actin fibers remained intact. On the other hand, ML-B completely disrupted the actin fibers distributed in both areas. Treatment of Vero cells with C-D or ML-B inhibited the replication of CDV. Double staining of CDV-infected Vero cells with antibody to N-protein and rhodamine-phalloidin revealed the presence of N-protein in mid-cytoplasm. However, the N-protein was specifically localized at the submembrane region in the presence of C-D, whereas it was clustered in the presence of ML-B. Viral mRNA levels of N- and H-proteins were rather increased by treatment with C-D or ML-B. The treatment with ML-B strongly inhibited N-protein expression, whereas C-D only slightly inhibited N-protein expression. These results suggest that actin microfilaments distributed in the cytoplasm and on the membrane region in host cells may have a different role in the process of CDV replication. PMID- 15133272 TI - Colonic ulceration and increase of neutrophil elastase activity in the acetic acid-induced colitis model in Syrian hamsters. AB - A novel colitis model using Syrian hamsters was developed. Colitis was induced by intracolonic administration of 1% acetic acid, and the ulcer area, tissue myeloperoxidase (MPO) activity, and luminal neutrophil elastase (NE) activity of the colon were determined at 1, 3, 8, 24 and 48 hr after colitis induction. The histopathological changes of the colon were also examined in this model. An increase of tissue MPO activity and NE activity was evident at 3 hr after induction of colitis, peaked at 24 hr, and decreased subsequently. The increase of luminal NE activity was well correlated with the colonic ulcer area. In histopathological examination, ulceration, erosion, crypt abscesses, neutrophil infiltration, hemorrhage, and edema were seen. The effects of prednisolone were examined to evaluate the adequacy of our colitis model. Syrian hamsters were treated orally with prednisolone at 18 and 1 hr before and at 6 hr after induction of colitis, and the ulcer area, tissue MPO activity, and luminal NE activity were evaluated at 24 hr after colitis induction. Prednisolone therapy had little effect on the tissue MPO activity. However, the NE activity of the prednisolone-treated group was significantly decreased. In addition, although prednisolone did not significantly decrease the ulcer area, a tendency toward decrease was noted. We conclude that this new model of experimental colitis in Syrian hamsters is useful for investigating the pathophysiology of colitis, especially useful for studying the relationship between colitis and NE activity. PMID- 15133274 TI - Calcinosis circumscripta on lingual muscles and dermis in a dog. AB - A 10-month old male Shi-tzu was presented with a week history of scurfy skin and whitish masses located in the ventral portion of tongue. Grossly, two whitish prominences with diameters of 3 to 4 mm were found on the underside of the tongue. Microscopically, in ulcerative epidermis, deposition of dense, amorphous granular basophilic calcium salts was separated by thin fibrous connective tissue containing mild inflammation. Many fascicles were characterized by replacement of degenerating myofibers with calcification. The precise cause of calcification could not be determined; however, it is interesting that two different lesions shown in calcinosis circumscripta and slight nutritional myopathy were also observed simultaneously only in the tongue. PMID- 15133273 TI - Anemia with chronic renal disorder and disrupted metabolism of erythropoietin in ICR-derived glomerulonephritis (ICGN) mice. AB - The ICR-derived glomerulonephritis (ICGN) mouse, a new inbred mouse strain with a hereditary nephrotic syndrome, is considered to be a good model of human idiopathic nephrotic syndrome and notably exhibits proteinuria and hypoproteinemia from the neonatal stage. In chronic renal disorder (CRD), anemia is a major subsequent symptom (renal anemia). The precise cause of renal anemia remains unclear, primarily owing to the lack of appropriate spontaneous animal models for CRD. To establish adequate animal models for anemia with CRD, we examined the hematological-biochemical properties and histopathological characteristics. With the deterioration of renal function, ICGN mice developed a normochromic and normocytic anemia, and exhibited normochromic and microcytic at the terminal stage. The expression of erythropoietin (EPO) mRNA both in the kidneys and liver and the EPO leak into the urine were observed in ICGN mice, indicating a disrupted metabolism of EPO in ICGN mice. In addition, a lack of iron induced by the hemolysis in the spleen and the leak of transferrin into urine as proteinuria aggravated the anemic condition. In conclusion, the ICGN mouse is a good model for anemia with CRD. PMID- 15133275 TI - Solitary nonepitheliotropic T-cell lymphoma in a dog. AB - A 7-year-old male Golden Retriever with swelling of the rostral bridge and right wing of the nasal areas, sneezing, and inspiratory difficulty was referred to a neighbor veterinarian. Except for those in the nasal area, no lesions were noted during routine physical examination. The mass occupying the nasal cavity was not observed radiographically. Punch biopsy of the affected lesions revealed nonepitheliotropic lymphoma. Immunohistochemical staining for CD3 antigen was positive. The dog was diagnosed with solitary nonepitheliotropic T-cell lymphoma. Local radiotherapy and systemic chemotherapy with doxorubicin were instituted and resulted in total clinical remission. The dog has remained disease free for 30 months. PMID- 15133276 TI - Serological evidence of akabane virus infection in northern Israel in 2001. AB - In February 2002 the first cases of a "blind newborn calves" syndrome with hydranencephaly appeared in Israel. Eighty-one serum samples, from 54 animals on farms where the syndrome was recorded and 27 others from unaffected farms were examined by neutralization of Akabane virus (AKAV, strain OBE-1) by the micro titer method. Forty-seven of the 54 samples from the affected farms contained high serum neutralization titers against AKAV (mean SN titer 79.5 and +/- 44.7, standard deviation), whereas only one of the 27 samples from the unaffected farms was positive (titer of 8). These results suggest that the vector(s) of AKAV was circulating in Israel in August through December, 2001. PMID- 15133277 TI - A primary hepatic plasma cell tumor in a dog. AB - An 8-year-old female Shetland sheep dog had hyperproteinemia with a monoclonal gammopathy and a solid mass on the liver, which was histologically diagnosed as a plasma cell tumor. After the treatment of surgery and chemotherapy, serum protein level reduced to the normal range and the gammopathy was disappeared. These findings indicate the plasma cell tumor developed primarily from the liver. PMID- 15133278 TI - A simple calculation for obtaining shunt fractions of portosystemic shunts. AB - Since the isotopes can not be utilized for veterinary patients in Japan, the authors developed a simple calculation formula of shunt fraction of portosystemic shunt based on the hepatic circulation model. The shunt fraction can be calculated utilizing only 2 portal pressure measurements of pre-shunt ligation and temporary or permanent shunt ligation. The calculated shunt fraction can obtained pre-ligation and post-ligation either temporally or permanent complete shunt ligation: complete ligation group of PSS (n=59) had 48.2 +/- 16.9% of shunt fractions, whereas the partial ligation group (n=48) had 71.6 +/- 10.7% of shunt fractions. PMID- 15133279 TI - Restriction of bovine herpesvirus 1 (BHV-1) growth in non-permissive cells beyond the expression of immediate early genes. AB - Mouse BALB/3T3-A31-1-1 (A31) cells are non-permissive to bovine herpes virus-1 (BHV-1) but permissive to pseudorabies virus (PrV). The promoter activity of the immediate early gene of BHV-1 (BICP4) was very weak when compared with that of PrV in A31 cells. Infectious BHV-1 genomic DNA co-transfected into A31 cells with plasmids expressing BICP4 and BICP0 by a strong promoter failed to yield any progeny virus. Growth of BHV-1 in non-permissible A31 cells is restricted in many phases of the growth. The fact that expression of BICP4 and/or BICP0 in A31 cells does not improve the yield of progeny virus from infectious BHV-1 genomic DNA suggests that some more growth restrictions exist beyond the expression of BHV-1 immediate early proteins. PMID- 15133280 TI - Mammary lesions associated with bovine herpesvirus type 4 in a cow with clinical mastitis. AB - Intranuclear eosinophilic inclusion bodies were seen in the lactiferous duct and sinus epithelium of mammary tissues collected from a cow with clinical mastitis. Transmission electron microscopy revealed herpesvirus particles in these cells. Immunolabeling against anti bovine herpesvirus type 4 (BHV-4) rabbit serum was detected in nuclei that had intranuclear inclusion bodies. In addition, BHV-4 was isolated from the mammary tissue. The viral DNA was detected by nested PCR from the same tissue. This is the first report to describe mammary lesions in association with BHV-4. PMID- 15133281 TI - Attachment and infection to MA104 cells of avian rotaviruses require the presence of sialic acid on the cell surface. AB - To determine the characters of receptors on target cells for avian rotaviruses, the receptors on MA104 cells for the pigeon rotavirus PO-13, the turkey rotaviruses Ty-1 and Ty-3, and the chicken rotavirus Ch-1 were analyzed. Pretreatment of MA104 cells with neuraminidase greatly reduced the infection by all of the four avian rotavirus strains. Binding of the cell-attachment protein, purified VP8 expressed in bacteria, of strain PO-13 to MA104 cells was also inhibited by pretreatment of cells with neuraminidase. These findings suggest that avian rotaviruses primarily utilize sialic acid-containing molecules as receptors on MA 104 cells. PMID- 15133282 TI - An abortion storm in cattle associated with neosporosis in Taiwan. AB - An abortion storm associated with acute neosporosis involving 18 cattle was observed in a dairy farm in Taiwan. Aborted fetus age ranged from 3 to 8 months. Of the 38 cattle in that farm examined during the abortion storm, 52.6% (20/38), 13.2% (5/38) and 10.5% (4/38) contained both IgG and IgM, only IgG and only IgM antibodies to Neospora caninum, respectively. No antibody to N. caninum was detected prior to the abortion storm. Follow-up study conducted a year later showed that 23 out of 28 cattle had sero-converted. Since some cattle were positive to either only IgG or IgM, we suggest that both IgG and IgM should be tested for diagnosing neosporosis. Neosporosis surveillance of naive cattle herd is recommended. PMID- 15133283 TI - Astronaut EVA exposure estimates from CAD model spacesuit geometry. AB - Ongoing assembly and maintenance activities at the International Space Station (ISS) require much more extravehicular activity (EVA) than did the earlier U.S. Space Shuttle missions. It is thus desirable to determine and analyze, and possibly foresee, as accurately as possible what radiation exposures crew members involved in EVAs will experience in order to minimize risks and to establish exposure limits that must not to be exceeded. A detailed CAD model of the U.S. Space Shuttle EVA Spacesuit, developed at NASA Langley Research Center (LaRC), is used to represent the directional shielding of an astronaut; it has detailed helmet and backpack structures, hard upper torso, and multilayer space suit fabric material. The NASA Computerized Anatomical Male and Female (CAM and CAF) models are used in conjunction with the space suit CAD model for dose evaluation within the human body. The particle environments are taken from the orbit averaged NASA AP8 and AE8 models at solar cycle maxima and minima. The transport of energetic particles through space suit materials and body tissue is calculated by using the NASA LaRC HZETRN code for hadrons and a recently developed deterministic transport code, ELTRN, for electrons. The doses within the CAM and CAF models are determined from energy deposition at given target points along 968 directional rays convergent on the points and are evaluated for several points on the skin and within the body. Dosimetric quantities include contributions from primary protons, light ions, and electrons, as well as from secondary brehmsstrahlung and target fragments. Directional dose patterns are displayed as rays and on spherical surfaces by the use of a color relative intensity representation. PMID- 15133284 TI - Prevention of gamma radiation induced anaemia in mice by diltiazem. AB - Intraperitoneal administration of diltiazem (DTZ), half an hour prior to whole body gamma irradiation (2.5, 5.0, & 7.5 Gy), showed the protection of animals from radiation-induced anaemia. Radiation exposure significantly (p < 0.001) reduced the number of pro- and normoblasts in bone marrow and RBC counts, hemoglobin (Hb), hematocrit (Hct ), and erythropoietin (EPO) level in blood, but increased myeloid / erythroid ratio. At all the radiation doses, the maximum decrease in these values was noted on the 3rd day, followed by a gradual recovery from the 7th day, but it was not recorded as normal even until the end of experimentation. In animals pretreated with DTZ, these values were measured higher at all the time periods in comparison to corresponding control, and these were almost normal at the last autopsy interval only at 2.5 Gy radiation dose. DTZ maintained the higher erythropoietin level in blood, which acted on bone marrow and spleen colony forming unit for erythroblast (CFU-E), and stimulated such cells to produce RBCs. These results confirm that DTZ has the potency to alter anaemic condition favorably through the protection of bone marrow stem cells, and subsequently it maintains the higher number of pro- and normoblasts in bone marrow, RBC counts, hemoglobin (Hb), hematocrit (Hct) percentage, and erythropoietin level in blood and the lower myeloid/erythroid ratio in bone marrow. PMID- 15133285 TI - Transplantation of dermal multipotent cells promotes the hematopoietic recovery in sublethally irradiated rats. AB - Our previous study indicated that dermal multipotent cells with the differentiation capacity to form cells with the phenotypic properties of osteocytes, adipocytes, chondrocytes, and neurons in specific inducing media could be isolated from the enzymatically dissociated dermal cells of newborn rats by their adherence to culture dish plastic. We have also observed that the systemic transplantation of dermal multipotent cells could not repopulate the hematopoietic system in lethally irradiated rats. In this paper, we found that a transplantation of plastic-adherent dermal multipotent cells into sublethally irradiated rats led to a significant increase of white blood cells in peripheral blood, nucleated cells, CFU-GM, and CFU-F colonies in bone marrow. FISH analysis, using a Y-chromosome specific probe, showed that dermal multipotent cells could engraft into bone marrow in recipients. Flow cytometry (FACS) analysis also showed that the proportion of CD2 and CD25 positive lymphocytes in peripheral blood did not change significantly in two weeks after transplantation. By these results, we infer that dermal multipotent cells may represent an alternative origin of mesenchymal stem cells to restore marrow microenvironment and promote the survival, engraftment, and proliferation of hematopoietic cells. PMID- 15133286 TI - Retrotransposition of limited deletion type of intracisternal A-particle elements in the myeloid leukemia Clls of C3H/He mice. AB - The murine genome has about 1,000 copies of DNA elements for the intracisternal A particle (IAP) that resembles a retrovirus. We previously reported that the genomic DNA of the cells from radiation-induced acute myeloid leukemia (AML) lines derived from C3H/He inbred mice was frequently rearranged by the integration of the IAP element. In this study, 8 IAP elements from the characteristic integration sites in 6 cell lines of radiation-induced AML from different mice were characterized and compared in structure with 114 IAP elements isolated from the normal C3H/He genome. One of the 8 elements was a full-length type I IAP, and 7 were of type-I Delta 1 with a common deletion site. Although the type I Delta 1 form is a minor population accounting for about 6% of total genomic IAP elements, it is predominantly retrotransposed in the AML cells from different C3H/He mice. This indicates that limited populations of the IAP elements contribute to the unique retrotransposition in AML cells. PMID- 15133287 TI - Low dose gamma-rays activate immune functions via induction of glutathione and delay tumor growth. AB - We examined whether the increase of glutathione level induced by low dose gamma ray irradiation is involved in the appearance of enhanced natural killer (NK) activity and antibody-dependent cellular cytotoxicity (ADCC), leading to delayed tumor growth in Ehrlich solid tumor-bearing mice. NK activity in ICR mouse splenocytes significantly increased from 4 h to 6 h after whole-body gamma-ray irradiation at 0.5 Gy, and thereafter decreased almost to the zero-time level by 24 h post-irradiation. ADCC also increased significantly in a similar way. Reduced glutathione exogenously added to splenocytes obtained from normal mice enhanced both NK activity and ADCC in a dose-dependent manner. Since immune functions were enhanced through the induction of cellular glutathione after low dose irradiation, the inhibitory effect of the radiation on tumor growth was then examined in Ehrlich solid tumor-bearing mice. Tumor growth after inoculation was significantly delayed by the radiation. These results suggest that low-dose gamma rays activate immune functions via an induction of glutathione, leading to a delay of tumor growth. PMID- 15133288 TI - Radiation and epidemiological analysis for solid cancer incidence among nuclear workers who participated in recovery operations following the accident at the Chernobyl NPP. AB - This paper discusses the results of the analysis of the relationship between dose and solid cancer incidence among nuclear workers (males) who worked as liquidators after the Chernobyl accident. Information on this cohort of individuals is available at the regional center of Russian National Medical and Dosimetric Registry operating at the RF State Research Centre-Institute of Biophysics. Medical and dosimetric information on 8,654 persons 18-60 years of age with documented external radiation doses is used for the analysis. These data were gathered in the period from 1996 to 2001 and cover a total of 45,166.5 follow-up person-years. In the cohort under study, 179 solid cancers occurred during this period. The average age of liquidators at the time of exposure was 35.8 years, and the average dose as a result of the Chernobyl exposure was about 0.05 Sv. For an analysis of the dose-effect relationship (induction of radiation induced malignant neoplasms) the statistical software EPICURE was used. The results of the analysis show that the cancer incidence in this cohort does not exceed cancer incidence in relevant age groups of the Russian population. The mean value of SIR for all cancer diseases was 0.88 (0.76, 1.02, 95% CI) for the whole period of follow-up. Risks for the induction of radiation-related cancer diseases were not statistically meaningful. Excess relative risk per 1 Sv was 0.95 (-1.52, 4.49, 95% CI). PMID- 15133289 TI - Experimental approach to improving early postirradiation restoration in the hemopoietic system of irradiated canines. AB - Experiments on mongrel canines exposed to total body irradiation in free-in-air dose of 3.85 (LD 95/45) or 4.05 Gy were carried out to evaluate the therapeutic effectiveness of some of bone marrow immediate-after-irradiation extraction, incubation in the presence of protein synthesis inhibitor and reimplantation in the organism (the reimplantation method) or bone marrow extraction alone (the extraction method). There were tested various kinds of methods that differed in bone marrow-blood mix volume extracted, in large or in small volumes. It has been determined that the reimplantation method in large volumes and the extraction method in small volumes are equally effective. Being supplemented with supportive therapy by antibiotics during d8-d24, the methods allowed the rescue at 4.05 Gy irradiation significantly more animals than in the case of therapy alone or its combination with ineffective variants of the methods. The positive effect of the methods manifested in a higher level of leukocyte nadir on d17, earlier reaching 0.5 x 10(9) and 1 x 10(9) leukocytes per liter, and increasing 45 days survival. The mechanism of positive influence of the methods on the radiation injury of hemopoesis seems to be related to increased cytokine producing because of the irritation of bone marrow stromal cells and thus favorable interference in the early restoration processes. PMID- 15133290 TI - Microarray analysis of temporal gene responses to ionizing radiation in two glioblastoma cell lines: up-regulation of DNA repair genes. AB - To determine the patterns of gene expression responsible for the radiosensitivity of glioblastoma cells, we analyzed transcriptional changes after ionizing radiation in different cell lines. After completing clonogenic survival assays, we selected two glioblastoma cell lines with different radiosensitivities. Subsequently, they were investigated by using the technique of DNA microarray, and we then categorized the upregulated genes into 10 groups. Between the two cell lines, the difference in the percentage of DNA repair/replication category was the largest, and this category was present at a greater percentage with radioresistant cell line U87MG. Moreover, among the commonly upregulated genes, the DNA repair/replication category was present in the largest percentage. These genes included G22P1 (Ku70) and XRCC5 (Ku80) genes known as important members of the nonhomologous end-joining (NHEJ) pathway of DNA double strand break (DSB) repair. Furthermore, cell line that specifically upregulated genes included the members of major pathways of DNA DSB or single strand damage repair. These pathways were not only NHEJ, but also homologous recombination (HR) and postreplication repair (PRR). In conclusion, the distribution of genes involved in the DNA repair/replication category was most different between two human glioblastoma cell lines of different radiosensitivities. Among commonly upregulated genes, the DNA repair/replication category was present in the largest percentage. PMID- 15133291 TI - Radioprotective potential of an herbal extract of Tinospora cordifolia. AB - A preparation of Tinospora cordifolia (RTc) administered i.p. (200 mg/kg b.w.) to strain "A" male mice 1 h before whole body gamma-irradiation was evaluated for its radioprotective efficacy in terms of whole body survival, spleen colony forming units (CFU), hematological parameters, cell cycle progression, and micronuclei induction. Preirradiation treatment with RTc rendered 76.3% survival (30 days), compared to 100% mortality in irradiated control and prevented radiation induced weight loss. On 10th postirradiation day, the endogenous CFU counts in spleen were decreased with increasing radiation doses 12.0 (5 Gy), 2.16 (7.5 Gy) and 0.33 (10 Gy) but pre-irradiation administration of 200 mg/kg b.w. of RTc increased CFU counts to 31.16, 21.83 and 3.00 respectively. Pre-irradiation RTc treatment could restore total lymphocyte counts (TLC) by the 15th day to normal. It also increased the S-phase cell population that was reduced following 2 Gy irradiation in a time dependent manner. 2 Gy irradiation-induced micronuclei were also decreased by a pre-irradiation administration of RTc from 2.9 to 0.52%. Because the radioprotective manifestation of RTc observed in several systems in experimental animals can be exploited for human applications. PMID- 15133292 TI - Comparative study on Tp53 gene mutations in lung tumors from rats exposed to 239Pu, 237Np and 222Rn. AB - The tumor suppressor gene Tp53 was analyzed by polymerase chain reaction amplification of genomic DNA extracted from paraffin-embedded tissue sections of rat lung tumors to compare mutations that occurred after inhalation exposures to plutonium dioxide, neptunium dioxide, or radon and radon progenies. Exons 5 to 8 of the gene were amplified in 16 plutonium-, 23 neptunium- and 15 radon-induced lung tumors, and their polymerase chain reaction products were examined for mutations by single strand conformational polymorphism analysis and direct sequencing method. Two point mutations were detected in the plutonium-induced tumors, i.e., a guanine to adenine transition at codon 219 of exon 6 and a cytosine to thymine transition at codon 266 of exon 8. Although only one point mutation was found at codon 175 of exon 5 (cytosine to thymine transition) from neptunium-induced tumors, no mutations were detectable from radon-induced tumors. These results indicate that the abnormalities of the Tp53 gene might not be so critical for the pulmonary carcinogenesis after the inhalation of different alpha emitters, even though the presence and frequencies of the Tp53 gene mutations were different. PMID- 15133293 TI - Properties of radicals formed by the irradiation of wool fibers. AB - Wool fibers of different sample conditions were irradiated in different atmospheres by (60)Co gamma-rays and were studied by electron spin resonance method (ESR). It was found that a large percentage of the alpha-carbon radicals of polymer main chain were more long-lived radicals. The ESR measurements of irradiated cortex samples of the wool fibers proved that most radicals from the cortex were long-lived ones. Low water content (as low as 27.5%) in the reaction system did not greatly affect the radical formation, but higher water contents would reduce the radical concentrations dramatically and accelerate their decaying process. The results will be of help in property modification of wool products by radiation graft copolymerization. PMID- 15133294 TI - Biochemical comparison between radon effects and thermal effects on humans in radon hot spring therapy. AB - The radioactive and thermal effects of radon hot spring were biochemically compared under a sauna room or hot spring conditions with a similar chemical component, using the parameters that are closely involved in the clinic for radon therapy. The results showed that the radon and thermal therapy enhanced the antioxidation functions, such as the activities of superoxide dismutase (SOD) and catalase, which inhibit lipid peroxidation and total cholesterol produced in the body. Moreover the therapy enhanced concanavalin A (ConA)-induced mitogen response and increased the percentage of CD4 positive cells, which is the marker of helper T cells, and decreased the percentage of CD8 positive cells, which is the common marker of killer T cells and suppressor T cells, in the white blood cell differentiation antigen (CD8/CD4) assay. Furthermore, the therapy increased the levels of alpha atrial natriuretic polypeptide (alpha ANP), beta endorphin, adrenocorticotropic hormone (ACTH), insulin and glucose-6-phosphate dehydrogenase (G-6-PDH), and it decreased the vasopression level. The results were on the whole larger in the radon group than in the thermal group. The findings suggest that radon therapy contributes more to the prevention of life-style-related diseases related to peroxidation reactions and immune suppression than to thermal therapy. Moreover, these indicate what may be a part of the mechanism for the alleviation of hypertension, osteoarthritis (pain), and diabetes mellitus brought about more by radon therapy than by thermal therapy. PMID- 15133295 TI - Inhibitory effects of prior low-dose X-ray irradiation on carbon tetrachloride induced hepatopathy in acatalasemic mice. AB - The catalase activities in blood and organs of the acatalasemic (C3H/AnLCs(b)Cs(b)) mouse of C3H strain are lower than those of the normal (C3H/AnLCs (a)Cs(a)) mouse. We examined the effects of prior low-dose (0.5 Gy) X ray irradiation, which reduced the oxidative damage under carbon tetrachloride induced hepatopathy in the acatalasemic or normal mice. The acatalasemic mice showed a significantly lower catalase activity and a significantly higher glutathione peroxidase activity compared with those in the normal mice. Moreover, low-dose irradiation increased the catalase activity in the acatalasemic mouse liver to a level similar to that of the normal mouse liver. Pathological examinations and analyses of blood glutamic oxaloacetic and glutamic pyruvic transaminase activity and lipid peroxide levels showed that carbon tetrachloride induced hepatopathy was inhibited by low-dose irradiation. These findings may indicate that the free radical reaction induced by the lack of catalase and the administration of carbon tetrachloride is more properly neutralized by high glutathione peroxidase activity and low-dose irradiation in the acatalasemic mouse liver. PMID- 15133296 TI - Bystander effect in lymphoma cells vicinal to irradiated neoplastic epithelial cells: nitric oxide is involved. AB - Evidence has been accumulated for attached cells demonstrating that nonirradiated cells can have a response to the ionization events delivered to their neighbors. In the present study, we first investigated the bystander responses between suspension and neoplastic cells by coculturing L5178Y (LY) cells with human salivary gland (HSG) cells that had been irradiated with either 290 MeV/u carbon ions or X-rays. After this coculture, the survival of nonirradiated recipient LY cells showed dichotomous responses to the irradiation dose delivered to HSG cells. Apoptosis and necrosis were also produced in a 48 h subculture of the recipient LY cells, and their yield increased, but then had a tendency to decrease when the irradiation dose increased. Treatment of cells with PTIO, a nitric oxide specific scavenger, diminished apoptosis and necrosis of the recipient LY cells to the control level. As an oxidization product of NO, nitrite was detected in the coculture medium and its time course corresponded well to the decrease of the viability of irradiated HSG cells. Moreover, the relationship of the survival and the apoptotic and necrotic production of the recipient LY cells to the nitrite concentration followed a linear-quadratic model. The present findings of NO being involved in the radiation-induced bystander effect may have significance in terms of radiotherapy. PMID- 15133297 TI - Delayed induction of telomere instability in normal human fibroblast cells by ionizing radiation. AB - We examined the delayed induction of telomere instability in hTERT-immortalized normal human fibroblast (BJ1-hTERT) cells exposed to X-rays. BJ1-hTERT cells were irradiated with 2 Gy of X-rays, and chromosome aberrations were analyzed 24 hours after irradiation and in the surviving cells 14 days after X-ray exposure. We found that the X-ray-surviving cells showed an increased frequency of chromatid gaps and breaks and chromosome fragments compared to the control cells. Furthermore, centromere- and telomere-FISH revealed that the frequency of telomere loss and duplication significantly increased in surviving cells compared to the control level. Because no induction of telomere abnormality was observed in cells 24 hours after irradiation, X-irradiation might not affect telomeres directly, but it specifically induces delayed telomere instability in normal human fibroblast cells. PMID- 15133298 TI - Effects of gemcitabine on cell survival and chromosome aberrations after pulsed low dose-rate irradiation. AB - The radiosensitizing potential of gemcitabine (2',2'-difluoro-2'-deoxycytidine) was studied in combination with pulsed low dose-rate irradiation. The experiments were carried out with a human lung carcinoma cell line SW1573. These were irradiated at pulsed low dose rate (p-LDR); the average dose rate was 1 Gy/h. In the experiments with gemcitabine, this drug was applied for 24 h at a concentration of 10 nM prior to irradiation. The response of the cells to treatment was tested by using the standard clonogenic assay. Next to the cell killing effects, damage to chromosomes was also assayed by using by whole chromosome Fluorescent In Situ Hybridization (FISH). Damage in chromosomes 2 and 18 was visualized by whole chromosome FISH and scored according to the PAINT method. A clear enhancement of the effects of radiation on cell survival was observed by preincubation of the cells with gemcitabine. The enhancement factor obtained from the p-LDR data was 1.7, which is much lower than the enhancement factor of 2.9 at high-dose rate. We did not observe a consistent increase in color junctions concomitant with radiosensitization. In chromosome 2, a small increase, and in chromosome 18, a decrease, in the number of color junctions was observed after radiation combined with gemcitabine compared to irradiation alone. These differences were not statistically significant. However, for the (unstable) acentric chromosome fragments from both chromosomes, significant changes were observed: In the case of chromosome 2, an increase, and in the case of chromosome 18, a decrease. So these results indicate that gemcitabine has no large and consistent effect on the repair of genomic lesions that induce secondary chromosome breaks. Although it is clear that gemcitabine-induced radiosensitization can be expected when it is combined with brachytherapy, as with radiation at a high-dose rate, the mechanism of radiosensitization is so far not evident, and further experiments will be needed to elucidate this. PMID- 15133299 TI - The establishment and characterization of cell lines stably expressing human Ku80 tagged with enhanced green fluorescent protein. AB - The Ku protein is a complex of two subunits, Ku70 and Ku80, and it plays a role in multiple nuclear processes, e.g., nonhomologous DNA-end-joining (NHEJ), chromosome maintenance, and transcription regulation. On the other hand, several studies have reported a cytoplasmic or cell surface localization of Ku in various cell types. The mechanism underlying the regulation of all the diverse functions of Ku is still unclear, though the mechanism that regulates the nuclear localization of Ku70 and Ku80 appears to play, at least in part, a key role in regulating the physiological function of Ku. In this study, we generated cell lines expressing the human Ku80 tagged with the green fluorescent protein (GFP) color variants in Ku80-deficient cells, i.e., xrs-6 derived from CHO-K1. Although Ku70, as well as Ku80, was undetectable in xrs-6 cells, it was seen in these transformants at a level similar to the level of CHO-K1. Furthermore, etoposide- and radiosensitive phenotype of xrs-6 cells were corrected by an introduction of the tagged Ku80. Moreover, the tagged Ku80 suppressed apoptosis triggered by DNA damage. These results demonstrate that fusion to the GFP color variants does not interfere with the functions of the Ku80 in the Ku-dependent DSB repair. Therefore, these transformants might be useful not only in the analysis of Ku80 behavior, but also in an analysis of the dynamics of the NHEJ repair process. PMID- 15133300 TI - Differential activation of kinases in ex vivo and in vivo irradiated mice lymphocytes. AB - Various kinases, such as tyrosine, protein kinase C (PKC) and MAP kinase, play important role in the cellular response to radiation, but little is known about the specific response in the whole animal. Most studies, except a few, are based on single cells. There is a paucity of data where signaling following whole body irradiation is concerned. In this study a comparison has been made between the activities of these kinases following ex vivo and in vivo irradiation. Tyrosine kinase activity showed no difference in the lymphocytes irradiated ex vivo or in vivo. A significant differential dose-dependent response could be observed in PKC activity. PKC was seen to be activated at the higher dose, i.e., 1 Gy in, in vivo irradiated lymphocytes, whereas in ex vivo irradiated lymphocytes, PKC was seen to be activated at the lower dose, i.e., 0.1 Gy. MAP kinase activity was seen to decrease with an increasing dose in ex vivo irradiated lymphocytes. In vivo MAP kinase activity was seen to increase as the dose increased, with maximum activation at 3 Gy. These kinases are being used to manipulate the tumor response to radiotherapy. Thus it is essential to study the behavior of the above kinases in the whole animal because the difference in response of a single cell to the whole animal may be different. PMID- 15133301 TI - Protective effect of an extract of Phyllanthus amarus against radiation-induced damage in mice. AB - The radioprotective effect of an extract of the plant Phyllanthus amarus (P. amarus) was investigated in adult BALB/c mice. P. amarus extract (750 mg/kg b.wt and 250 mg/kg b.wt) was administered orally to mice for five days prior to whole body radiation (6 Gy) and for one month after radiation. The animals were sacrificed on days 3, 9, 12, and 30 after radiation. P. amarus significantly increased the total W.B.C count, bone marrow cellularity, and alpha-esterase activity as compared to untreated radiation-exposed animals. P. amarus treatment also increased the activity of various antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST), glutathione peroxidase (GPX), and glutathione reductase (GR), both in blood and tissue, which were reduced by radiation treatment. There was also a significant increase in the glutathione (GSH) levels of blood and tissue. Lipid peroxidation levels, which were increased after radiation, were significantly reduced by P. amarus treatment, both in serum and liver. The results collectively indicate that P. amarus extract could increase the antioxidant defense mechanism in mice and there by protect the animals from radiation-induced cellular damage. PMID- 15133302 TI - Insulin-like growth factor I receptor does not contribute to heat shock-induced Activation of Akt and extracellular signal-regulated kinase (ERK) in mouse embryo fibroblasts. AB - We have investigated the role of insulin-like growth factor I receptor (IGF-IR) in heat shock-induced activation of the mitogen-activated protein kinase (MAPK) and phosphatidylinositol-3 ' kinase (PI3-K) pathways. We utilized mouse embryo fibroblasts (MEFs) devoid of endogenous IGF-IR (R-) and MEFs overexpressing human IGF-IR (WT) and examined the activation kinetics of extracellular signal regulated kinase (ERK) and Akt following heat shock treatment. There were no differences in the kinetics or temperature dependence of activation of either ERK or Akt between the cell lines. As expected, heat shock failed to induce autophosphorylation of IGF-IR overexpressed in WT cells. Surprisingly, the autophosphorylation of endogenous epidermal growth factor receptor (EGFR), which is thought to play an important role in heat shock-induced activation of the MAPK and PI3-K pathways, was not observed in either WT or R-cells. These results suggest that neither IGF-IR nor EGFR contributes to the heat shock -induced activation of ERK and Akt in these cell lines. PMID- 15133303 TI - The role of Tec family kinases in myeloid cells. AB - Members of the Tec kinase family (Bmx, Btk, Itk, Rlk and Tec) are primarily expressed in the hematopoietic system and form, after the Src kinase family, the second largest class of non-receptor protein tyrosine kinases. During lymphocyte development and activation Tec kinases have important functions in signaling pathways downstream of the antigen receptors. Tec family kinases are also expressed in cells of the myeloid lineage. However, with the exception of mast cells and platelets, their biological role in the myeloid system is only poorly understood. This review summarizes the current knowledge about the function of Tec family kinases in hematopoietic cells of the myeloid lineage. PMID- 15133304 TI - IL-9 in allergic inflammation. AB - Th2 type cytokines such as interleukin (IL)-4, IL-5, IL-9, and IL-13 are important mediators in allergic inflammation. The present review will focus on the role of IL-9 in allergic inflammation. The structure and genomic architecture of IL-9 and its receptor, the source of IL-9 and its regulation as well as its effects on different cell types will be reviewed. Furthermore, the specific role of IL-9 in allergic diseases and the potential therapeutic approach of blocking IL-9 will be discussed. PMID- 15133305 TI - Direct gliadin cytotoxicity as a cofactor in the pathogenesis of celiac disease. PMID- 15133306 TI - Combined adrenal myelolipoma and medullary hyperplasia. AB - OBJECTIVE: A patient is reported with hypertension due to combined medullary adrenal hyperplasia and myelolipoma. METHODS: A 52-year-old woman with long standing hypertension was evaluated for an incidentally discovered large tumor of the left adrenal. Left adrenalectomy was performed for a presumptive clinical diagnosis of pheochromocytoma. RESULTS: Histopathologic examination revealed a mixed tumor consisting of a large myelolipoma with infiltrating foci of adrenal medulla. CONCLUSIONS: A patient is described with hypertension, myelolipoma and adrenal medullary hyperplasia; following adrenalectomy, however, blood pressure and biochemical abnormalities normalized. PMID- 15133307 TI - Selecting loop breakers in general pedigrees. AB - The presence of loops in pedigrees poses severe computational problems in likelihood calculation that can be solved by creating an equivalent unlooped pedigree. We introduce a heuristic polynomial-time dynamic-programming algorithm, called SFH, that addresses the problem of selecting a minimal-cost set of loop breakers. We report computational experiments on simulated pedigrees with up to 1000 individuals and 361 loops, and multiple marriages. We compare the loop breaker set selected by our method with that obtained using the software package FASTLINK 4.1P. Our approach outperforms FASTLINK 4.1P on the computational-time point of view, on the point of view of quality of the loop-breaker set obtained, and on the point of view of the size of the problem that can be addressed. PMID- 15133309 TI - Power loss for linkage analysis due to the dichotomization of trichotomous phenotypes. AB - OBJECTIVES: Some traits, while naturally polychotomous, are routinely dichotomized for genetic analysis. Dichotomization, intuitively, leads to a loss of power to detect linkage, as some phenotypic variability is discarded. This paper examines this power loss in the context of a trichotomous trait. METHODS: To examine this power loss, we performed a simulation study where a trichotomous trait was simulated in a sample of 1,000 sib-pairs under various genetic models. The study was replicated 1,000 times. Linkage analysis using a variance components method, as implemented in Mx, was then performed on the trichotomous trait and compared with that on a dichotomized version of the trait. RESULTS: A comparison of the power and false positive rates of the analyses shows that power to detect linkage was increased by up to 22 percentage points simply by examining the trait as a trichotomy instead of a dichotomy. Under all models examined, the trichotomous analysis outperformed the dichotomous version. CONCLUSIONS: Comparable levels of false positive rates under both methods confirm that this power gain comes solely from the information lost upon dichotomization. Thus, dichotomizing tri- or poly-chotomous traits can lead to crippling power loss, especially in the case of many loci of small effect. PMID- 15133308 TI - Examination of potential overlap in autism and language loci on chromosomes 2, 7, and 13 in two independent samples ascertained for specific language impairment. AB - Specific language impairment is a neurodevelopmental disorder characterized by impairments essentially restricted to the domain of language and language learning skills. This contrasts with autism, which is a pervasive developmental disorder defined by multiple impairments in language, social reciprocity, narrow interests and/or repetitive behaviors. Genetic linkage studies and family data suggest that the two disorders may have genetic components in common. Two samples, from Canada and the US, selected for specific language impairment were genotyped at loci where such common genes are likely to reside. Significant evidence for linkage was previously observed at chromosome 13q21 in our Canadian sample (HLOD 3.56) and was confirmed in our US sample (HLOD 2.61). Using the posterior probability of linkage (PPL) to combine evidence for linkage across the two samples yielded a PPL over 92%. Two additional loci on chromosome 2 and 7 showed weak evidence for linkage. However, a marker in the cystic fibrosis transmembrane conductance regulator (7q31) showed evidence for association to SLI, confirming results from another group (O'Brien et al. 2003). Our results indicate that using samples selected for components of the autism phenotype may be a useful adjunct to autism genetics. PMID- 15133310 TI - Multilocus analysis of hypertension: a hierarchical approach. AB - While hypertension is a complex disease with a well-documented genetic component, genetic studies often fail to replicate findings. One possibility for such inconsistency is that the underlying genetics of hypertension is not based on single genes of major effect, but on interactions among genes. To test this hypothesis, we studied both single locus and multilocus effects, using a case control design of subjects from Ghana. Thirteen polymorphisms in eight candidate genes were studied. Each candidate gene has been shown to play a physiological role in blood pressure regulation and affects one of four pathways that modulate blood pressure: vasoconstriction (angiotensinogen, angiotensin converting enzyme ACE, angiotensin II receptor), nitric oxide (NO) dependent and NO independent vasodilation pathways and sodium balance (G protein-coupled receptor kinase, GRK4). We evaluated single site allelic and genotypic associations, multilocus genotype equilibrium and multilocus genotype associations, using multifactor dimensionality reduction (MDR). For MDR, we performed systematic reanalysis of the data to address the role of various physiological pathways. We found no significant single site associations, but the hypertensive class deviated significantly from genotype equilibrium in more than 25% of all multilocus comparisons (2,162 of 8,178), whereas the normotensive class rarely did (11 of 8,178). The MDR analysis identified a two-locus model including ACE and GRK4 that successfully predicted blood pressure phenotype 70.5% of the time. Thus, our data indicate epistatic interactions play a major role in hypertension susceptibility. Our data also support a model where multiple pathways need to be affected in order to predispose to hypertension. PMID- 15133311 TI - Percentiles of the null distribution of 2 maximum lod score tests. AB - We here consider the null distribution of the maximum lod score (LOD-M) obtained upon maximizing over transmission model parameters (penetrance values, dominance, and allele frequency) as well as the recombination fraction. Also considered is the lod score maximized over a fixed choice of genetic model parameters and recombination-fraction values set prior to the analysis (MMLS) as proposed by Hodge et al. The objective is to fit parametric distributions to MMLS and LOD-M. Our results are based on 3,600 simulations of samples of n = 100 nuclear families ascertained for having one affected member and at least one other sibling available for linkage analysis. Each null distribution is approximately a mixture p(2)(0) + (1 - p)(2)(v). The values of MMLS appear to fit the mixture 0.20(2)(0) + 0.80chi(2)(1.6). The mixture distribution 0.13(2)(0) + 0.87chi(2)(2.8). appears to describe the null distribution of LOD-M. From these results we derive a simple method for obtaining critical values of LOD-M and MMLS. PMID- 15133312 TI - Powerful allele sharing statistics for nonparametric linkage analysis. AB - Nonparametric linkage analysis is widely used to map susceptibility genes for complex diseases. This paper introduces six nonparametric statistics for measuring marker allele sharing among the affected members of a pedigree. We compare the power of these new statistics and three previous statistics to detect linkage with Mendelian diseases having recessive, additive, and dominant modes of inheritance. The nine statistics represent all possible combinations of three different IBD scoring functions and three different schemes for sampling genes among affecteds. Our results strongly suggest that the statistic T(rec)(blocks) is best for recessive traits, while the two statistics T(kin)(pairs) and T(all)(kin) vie for best for an additive trait. The best statistic for a dominant trait is less clear. The statistics T(kin)(pairs) and T(all)(kin) are equally promising for small sibships, but in extended pedigrees the statistics T(dom)(blocks) and T(dom)(pairs) appear best. For a complex trait, we advocate computing several of these statistics. PMID- 15133313 TI - Weakness of anastomotic site in diabetic rats is caused by changes in the integrity of newly formed collagen. AB - AIMS: Diabetes mellitus often causes anastomotic leakage leading to severe complications after surgery. The purpose of this study was to determine whether collagen synthesis is correlated with the anastomotic strength in diabetic animals in special reference to collagen type differences. METHODS: Thirty Lewis rats weighing approximately 250 g were made diabetic by tail vein injection of streptozotocin (STZ). Laparotomy was performed through a midline incision under ether anesthesia. A colonic segment 5 cm distal to the ileocecal junction was transected then anastomosed with absorbable suture. Anastomotic strength was measured on days 3, 5, 7 and 14 postoperatively, and compared with normal rats. Collagen type I and type III mRNA together with the total collagen concentration were measured in anastomotic sites. RESULTS: Anastomotic strength in diabetic rats was significantly lower on day 7, but gradually recovered up to day 14. There was no statistical significance between the two groups in hydroxyproline concentrations. Although the expressions of mRNA of collagen type III in both groups were statistically not different, that of collagen type I in the diabetes group merely increased and was clearly less than that of the control group. CONCLUSIONS: Anastomotic integrity in diabetic rats was significantly damaged on day 7 after surgery but recovered by day 14. This is caused not by the total contents of collagen accumulation but by the decrease of newly formed collagen type I contents. PMID- 15133314 TI - Evolution of plasma D-dimer and fibrinogen in witnessed onset of paroxysmal atrial fibrillation. AB - BACKGROUND: Although increased D-dimer and fibrinogen have been proved to be related with atrial fibrillation (AF), their evolution in the course of time remains unclear. METHODS: To elucidate the evolution of plasma D-dimer and fibrinogen in AF of different duration. 56 patients (group A) of 3,524 patients in whom the onset of AF had been witnessed in a chest pain clinic were enrolled for study. Plasma D-dimer and fibrinogen concentrations were checked within 30 min after the onset of AF and followed up as scheduled. Another 50 patients (group B) with chronic AF underwent the same protocol and served as controls. RESULTS: In group A, the D-dimer levels reached a plateau at the 18th hour (382 +/- 52 vs. 840 +/- 280 ng/ml, p < 0.001 by ANOVA) and the fibrinogen level increased gradually from 4.1 +/- 0.6 g/ml at baseline to 6.1 +/- 0.9 g/ml at the 48th hour (p < 0.001). No subjects had evidence of intra-cardiac thrombi by transesophageal echocardiography. There were significant differences in plasma D dimer and fibrinogen levels between the two groups at each measurement. At the cut- off value of 500 ng/ml, plasma D-dimer had a sensitivity of 100%, a specificity of 93% in defining AF lasting for less than 12 h. The positive and negative predictive values were 52% and 100%, respectively. Plasma D-dimer increased above the normal range prior to the 12th hour after the onset of AF. CONCLUSION: The observations support its use as a screening tool for identifying patients with short duration AF capable of being safely converted. PMID- 15133315 TI - Coenzyme Q10 levels in maternal plasma and cord blood:correlations with mode of delivery. AB - BACKGROUND: Oxygen (O2) plays a critical role in the O2-reduction reactions indispensable for life, but can produce free radicals that are involved in many diseases. Coenzyme Q10 (CoQ10), acting as a redox carrier in the respiratory chain, occupies a central position in the energy metabolism and oxidative defence. Neonates seem to be very subjected to oxidative stress because of their deficient antioxidant systems. DESIGN/METHODS: The aim of the study was to verify whether the mode of delivery may affect CoQ10 levels in the mother and neonate, and thus influence the risk of oxidative damage in the newborn. We measured CoQ10 levels in maternal plasma and cord blood at birth after three different modes of delivery (45 term healthy pregnancies): (1) vaginal in room air (VD) (n = 15); (2) elective caesarean section with general anaesthesia (50% O2 and 50% N2O) (CSg) (n = 15), and (3) elective caesarean section with spinal anaesthesia without O2 (CSs) (n = 15). Our results showed higher levels of Q10 in mothers and neonates with VD (1.29 +/- 0.43 and 0.15 +/- 0.06 microg/ml, respectively) or CSs (1.15 +/- 0.28 and 0.24 +/- 0.06 microg/ml, respectively) when compared to CSg (0.74 +/- 0.28 and 0.07 +/- 0.03 microg/ml, respectively) (p < 0.01). CONCLUSIONS: These data demonstrate that the mode of delivery may affect CoQ(10) levels in mothers and neonates, and thus influence the risk of oxidative damage in the newborn. PMID- 15133316 TI - Mineral and vitamin status in elderly persons from Northwest Spain consuming an Atlantic variant of the Mediterranean diet. AB - AIM: To assess dietary intake and serum mineral and vitamin levels in elderly people from Northwest Spain consuming a Mediterranean diet, rich in vegetables, fruit, meat, fish, olive oil, dairy products and moderate in wine. METHODS: Cross sectional observational study in four retirement homes. Forty-five men and 65 women participated. Dietary intake and serum calcium, magnesium, iron, retinol, alpha-tocopherol, albumin, cholesterol, glucose and hematological parameters were determined. RESULTS: Mean consumption of fruit plus vegetables was 600 g/day. Men consumed significantly more legumes, fruit, meat, and alcoholic beverages, but fewer vegetables than women. Women presented higher carbohydrate and lower alcohol energy contributions. Men consumed significantly more thiamin, niacin equivalents, riboflavin, retinol equivalents and iron. Vitamin D intake was 2.2 +/- 1.2 microg/day and folate intake was 204 +/- 47 microg/day without gender differences. Prevalence of anemia was 6.7% and that of high ferritin, 1.8%. Serum cholesterol and retinol were higher in women. Mean serum alpha tocopherol/cholesterol ratio in the whole population was 6.3 +/- 2.3 mmol/mol without gender differences. CONCLUSION: This elderly population consumes an Atlantic-Mediterranean diet that appears, according to biochemical and hematological parameters, appropriate. However, more fatty fish and leafy green vegetables and the inclusion of fortified foods in the diet might optimize micronutrient status. PMID- 15133317 TI - Influence of dietary habits on serum selenium concentration. AB - BACKGROUND: A serum selenium concentration of 70 microg/l is considered to be sufficient for glutathione peroxidase (GSHPx) activity, a selenium-dependent enzyme reflecting the body selenium status. The purpose of the study was to assess which foods or food groups best affect serum selenium in subjects with selenium concentrations below or above 70 microg/l. METHODS: A food frequency questionnaire was employed to describe the dietary habits of 129 subjects. Serum selenium concentrations were determined using the electrothermal absorption spectrometry (ETAAS) technique following serum dilution. RESULTS: Multiple regression analysis revealed that consumption of ham, honey and tea was positively associated with the selenium concentration in the sera of selenium inadequate subjects. In selenium-inadequate subjects, alcohol consumption may be responsible for selenium losses. Unlike the selenium-inadequate subjects, the selenium concentrations in the sera of the selenium-adequate subjects were positively correlated with the frequencies of wholegrain bread consumption and processed fruit consumption. In all the groups tested, the serum selenium concentration was not affected by cigarette smoking. CONCLUSIONS: The serum selenium concentration is influenced by dietary habits, but not by cigarette smoking. Frequent consumption of ham, tea and honey may be effective to improve the selenium concentration in the sera of selenium-inadequate subjects. PMID- 15133318 TI - Effects of dietary intake and life style on bone density in patients with diabetes mellitus type 2. AB - OBJECTIVE: The aim of our study was to investigate the relation among glycemic control, lifestyle and dietary intake with bone mineral density in patients with diabetes mellitus type 2. DESIGN: Cross-sectional study. SETTING: Tertiary care hospital. PARTICIPANTS: A cross-sectional study in a tertiary care hospital was performed. Ninety-two patients attending our diabetes service (56 females/36 males) with diabetes mellitus type 2 were enrolled in a consecutive way. The inclusion criteria were diabetes diagnosed >40 years of age, with type 2 diabetes defined in accordance with the criteria of the American Diabetes Association and no use of dietary supplements. Body mass index, waist-to-hip ratio, glucose level, and HbA1c levels were assessed in all patients. X-ray densitometry of the calcaneal region and a 3-days written food record keeping, and a qualitative questionnaire of lifestyle were also performed. RESULTS: A total of 21.7% of patients had osteoporosis (T score <2.5 SD). Patients were overweight with a high BMI and a medium glucose control. Patients with osteoporosis were older than those without osteoporosis (67.8 +/- 6.9 vs. 62.1 +/- 9.2 years; p < 0.05). Significant differences were detected between patients without and with osteoporosis in calcium intake (1,219.37 +/- 387 vs. 839 +/- 251 mg/day; p < 0.05) and zinc intake (9.23 +/- 3.5 vs. 13.3 +/- 6.9 mg/day; p < 0.05), respectively. No differences were detected in other dietary dairy intakes. In correlation analysis age (r = -0.23; p < 0.05) and BMI (r = 0.48; p < 0.05) was correlated with BMD. In univariate analysis with dicotomic variables, only exercise was positive associated with osteoporotic status (87.5% exercise habit in patients without osteoporosis and 25% exercise habit in patients with osteoporosis; p < 0.05). In a logistic model with the dependent variable (osteoporosis), remained in the final model dietary dairy intake of calcium and zinc, BMI, age and exercise. Exercise, calcium intake and BMI were protective factors. Zinc intake, and age were risk factors. CONCLUSIONS: Exercise, calcium intake, body mass index had a protective role in bone mineral density in patients with diabetes mellitus type 2. Zinc intake and age were risk factors in our population. PMID- 15133319 TI - Relationship between alpha-tocopherol content in the different lipoprotein fractions in term pregnant women and in umbilical cord blood. AB - AIMS: To determine the concentration of vitamin E in normal maternal and umbilical cord blood pairs, and to study the relationship between vitamin E content in maternal lipoprotein fractions and umbilical cord blood. METHODS: Fifty healthy pregnant women were recruited randomly at term and blood samples were drawn from the mothers at delivery and cord blood was obtained immediately postpartum. Vitamin E was determined by HPLC in plasma, in the different lipoprotein fractions and in the placenta. Plasma levels of triglycerides and cholesterol were also measured. RESULTS: The concentration of vitamin E in umbilical cord plasma was 250 microg/dl, lower than in maternal plasma (1,460 microg/dl) (p < 0.001). A positive correlation was found between the vitamin E concentration in maternal plasma, LDL and VLDL and in the umbilical cord plasma. In contrast, no correlation was found between maternal HDL concentration and umbilical cord blood. CONCLUSION: These results show that the concentration of vitamin E in umbilical cord blood is lower than in maternal plasma. LDL and VLDL seem to be the main source of vitamin E for the fetus. PMID- 15133320 TI - Effects of green tea catechin on polymorphonuclear leukocyte 5'-lipoxygenase activity, leukotriene B4 synthesis, and renal damage in diabetic rats. AB - The purpose of this study was to investigate the effects of green tea catechin on polymorphonuclear leukocyte 5'-lipoxygenase activity, leukotriene B4 synthesis, and renal damage in diabetic rats. Male Sprague-Dawley rats weighing 100 +/- 10 g were randomly assigned to 1 normal group and 3 diabetic groups given a catechin free diet (DM-0C group), 0.25% catechin diet (DM-0.25C group), or 0.5% catechin diet (DM-0.5C group), respectively. 5'-Lipoxygenase activity in the polymorphonuclear leukocytes significantly increased by 54% in the DM-0C group compared to the normal group, while the level in the DM-0.5C group remained the same as in the normal group. The leukotriene B4 content in the polymorphonuclear leukocytes increased 55% in the DM-0C group compared to the normal group, whereas the DM-0.25C and DM-0.5C groups exhibited the same level as the normal group. The superoxide radical content in the kidney microsomes increased 116% in the DM-0C group when compared to the normal group, yet decreased 29% in the DM-0.25C group and 50% in the DM-0.5C group compared to DM-0C group. The lipofuscin content was 197 and 136% higher in the DM-0C and DM-025C groups, respectively, than in the normal group, whereas the DM-0.5C group exhibited the same content as in the normal group. The carbonyl value increased 118% in the DM-0C group compared to the normal group, and the DM-0.25C and DM-0.5C groups were not significantly different from the DM-0C group. Accordingly, these results indicate that dietary catechin inhibited the generation of superoxide radicals, oxidized protein, and lipid peroxide in the kidney of streptozotocin-induced diabetic rats. Furthermore, green tea catechin supplementation in diabetic rats also appeared to inhibit the production of leukotriene B4 based on regulating the activity of 5' lipoxygenase, thereby potentially reducing renal oxidative damage and inflammatory reactions. PMID- 15133321 TI - Serum carotenoid and retinol levels during childhood infections. AB - BACKGROUND: In recent years, new physiological functions of vitamin A have been identified, including its role in immune defense. The antioxidant potential of carotenoids is thought to account for their health benefits. The aim of this study was to investigate the concentration changes in the provitamin-A (alpha- and-beta-carotene, beta-cryptoxanthin) and of the non-provitamin-A carotenoids (lutein, zeaxanthin and lycopene) in children with acute infections. METHODS: Serum retinol and carotenoid concentrations were measured by HPLC in 45 children suffering from acute infections and in 29 healthy controls. In addition 15 healthy children collected food samples over 2 days. C-reactive protein (CRP) levels were measured by turbidimetry. RESULTS: Serum levels of beta-carotene, alpha-carotene, beta-cryptoxanthin and retinol and non-provitamin-A carotenoid lutein and zeaxanthin concentrations were significantly lower in children with infections compared to healthy controls. There were significant negative correlations between CRP levels and serum beta-carotene and retinol concentrations. The average intake of retinol equivalents was 1.1 +/- 0.6 mg/day. CONCLUSION: Serum retinol and provitamin-A as well as non-provitamin-A carotenoid concentrations were lower in children with acute phase infections compared to healthy controls. The inverse correlations between CRP and beta-carotene or retinol indicate either decreased synthesis or increased utilization of these antioxidants. PMID- 15133322 TI - LDL susceptibility to copper-induced oxidation after administration of a single dose of free or esterified beta-cryptoxanthin. AB - BACKGROUND: The oxidative modification of LDL is believed to be an initial step in atherosclerosis. Thus, antioxidative substances such as carotenoids may have a role in the prevention of coronary heart disease. We examined the susceptibility of LDL to Cu2+ oxidation in young adults before and after a single dose of beta cryptoxanthin. METHODS: 1.3 mg of beta-cryptoxanthin was administered to 12 apparently healthy young volunteers. Six of the volunteers received esters, the other six free beta-cryptoxanthin. The plasma concentration of beta-cryptoxanthin and the susceptibility of LDL to copper-induced oxidation ex vivo in terms of the duration of lag time were measured before and 12 h after beta-cryptoxanthin ingestion. RESULTS: A single dose of beta-cryptoxanthin significantly increased the mean plasma beta-cryptoxanthin concentration and the mean cholesterol adjusted beta-cryptoxanthin concentration by 117 and 133%, respectively. No effect on the length of lag time was assessed. However, in LDL isolated from plasma 12 h after beta-cryptoxanthin administration the lengths of lag time correlated significantly with the plasma beta-cryptoxanthin concentration and with the cholesterol adjusted beta-cryptoxanthin levels. The lag time did not differ significantly between volunteers who received esters and those who received the same dosage as free beta-cryptoxanthin. At both measuring points, smokers, male volunteers and women using oral contraceptives tended to exhibit lower beta-cryptoxanthin concentrations and lower cholesterol adjusted beta cryptoxanthin concentrations as well as increased LDL oxidizability compared to nonsmokers and women not using oral contraceptives. CONCLUSION: A single dose of beta-cryptoxanthin does not enhance the duration of LDL lag time ex vivo in healthy young subjects. PMID- 15133323 TI - Effect of concomitant administration of cyclosporine and ketoconazole in children with focal segmental glomerulosclerosis. AB - BACKGROUND: Focal segmental glomerulonephritis (FSGS) is now the most common primary glomerulonephritis that leads to end-stage renal disease in both adults and children. Cyclosporine (CsA) is a well-known and effective immunosuppressive agent that has become a cornerstone of immunotherapy in solid organ transplantation and it has been used in the treatment of FSGS for over 15 years. The deliberate use of ketoconazole (keto) to reduce the need for CsA is not new, but it is particularly relevant because of the high cost of CsA. Many studies have documented this benefit in renal and cardiac transplants, but this co administration has not been reported in children with nephrotic syndrome (NS). METHODS: This study included 116 children (below 18 years of age) with primary FSGS who were steroid resistant or dependent and received CsA therapy. Among them, 88 received daily keto therapy (keto group) in a dose of 50 mg with concomitant decrease of CsA dose by one third, while 28 patients received CsA alone (non-keto group). Mean (+/-SD) age was 6.17 +/- 4.68 years and male to female ratio was 1.9:1. The great majority of the study population received the drugs for 1-2 years. The characteristics of both groups were comparable. RESULTS: Co-administration of keto significantly reduced mean doses of CsA by 46% at 1 year with overall net cost savings of 36%. It also significantly improved the response to CsA therapy and decreased the frequency of renal impairment. No significant side effects for keto were observed. CONCLUSION: Co-administration of keto and CsA in idiopathic FSGS children is safe. This combination not only reduces the costs but also may improve the response to CsA and stabilize the renal function. PMID- 15133324 TI - Epidemiology of nephrolithiasis today. AB - The epidemiology of nephrolithiasis differs according to geographical area and socio-economic conditions. In Italy the prevalence of the disease in 1993 was 17.2/1,000 inhabitants, most patients are men and elderly. The relative risk increased in subjects with family history for calculosis, with the tendency to eat protein-rich food and with overweight and body mass index (MBI) >32 kg/m2. Calcium oxalate and/or phosphate stones account for almost 70% of all renal stones observed in economically developed countries. PMID- 15133325 TI - Crystallization inhibitors in the pathophysiology and treatment of nephrolithiasis. AB - It is currently agreed that stone formation in the urinary tract requires supersaturation with respect to a given solid phase. However, this principle fully applies only to stones other than calcium-containing stones, in which case compounds acting as inhibitors are postulated to naturally occur in urine. Stone formation would therefore ensue from an imbalance between promoters and inhibitors. The saturation state can be estimated by means of computer model systems based on ab initio calculations, which account for the main soluble complexes formed in urine between relevant cations and anions. This estimates the overall promoting potential of urine. However, in the case of calcium nephrolithiasis, supersaturation does not make a clear-cut separation between normal subjects and patients. Several studies in the last two decades have identified many inhibitors of calcium oxalate and calcium phosphate crystallization, which are classified into the ionic and macromolecular. They have been shown to act on kinetics by interfering with nucleation, growth and aggregation of crystals. Unfortunately, except for citrate, none of the newly discovered substances has been definitely characterized in its molecular composition and structure, type and potency of inhibition, differences in concentration and structure between stone-forming and non stone-forming subjects. Citrate exhibits a dual action in urine, opposing crystal formation by both thermodynamic and kinetic mechanisms. At present it is the only natural inhibitor which can be measured in urine, quantitated as to inhibitory activity and used in medical treatment. PMID- 15133326 TI - Genetics of primary hyperparathyroidism. AB - Primary hyperparathyroidism, a genetically heterogeneous disease, usually occurs as a sporadic disorder due to the presence of parathyroid adenoma/s, hyperplasia or, rarely, carcinoma. In the last decades familial forms of primary hyperparathyroidism have been described. Recognizing such forms is essential for a correct clinical management of affected individual subjects and families. In fact, primary hyperparathyroidism may be the typical feature of familial syndrome or alternatively only an associated disorder within the context of a more complex syndromic picture. Several responsible genes have been so far identified, making their mutational analysis possible, which provides not only early identification of asymptomatic gene carriers, but could also add new important knowledge of the molecular mechanisms underlying parathyroid tumorigenesis. Such mechanisms could, in the near future, become an ideal target for new therapeutic strategies of primary hyperparathyroidism. PMID- 15133327 TI - Osteoporosis and urolithiasis. AB - Several studies have indicated that up to 60% of idiopathic calcium stone formers present hypercalciuria. Many authors have described reduced bone mineral density (BMD) in stoneformers with hypercalciuria, but osteopenia has also been found in normocalciuric patients. Moreover, Jaeger's group found that bone mass was reduced in all patients with calcium stone disease, independently of hypercalciuria. Many factors may contribute to the pathogenesis of osteopenia in stone formers. A predominant role has been given to the low-calcium diet that is still prescribed in nephrolithiasis. Also slight metabolic acidosis, which is frequently present in stone formers eating a diet rich in animal protein, can contribute to bone loss. Finally, some authors described a pathogenetic role for cytokines, prostaglandins and vitamin D receptor gene polymorphisms. PMID- 15133328 TI - Pathophysiology of reno-ureteral colic. AB - INTRODUCTION: Reno-ureteral colic presents with a complex of acute symptoms that usually indicate the presence of a stone in the collecting system, The aim of any treatment is pain relief, stone removal and preservation of kidney function. We focused our review on the pathophysiological aspects of reno-ureteral colic and, briefly, on its treatment. MATERIALS AND METHODS: A complete overview of contemporary literature regarding reno-ureteral colic was performed by using MEDLINE and website on-line publications like EAU Guidelines. RESULTS: Clinical presentation of reno-ureteral colic, of mechanisms of ureteral peristalsis and pathophysiology of renal colic is described. An overview of the available medical treatments is presented. CONCLUSIONS: An accurate comprehension of the pathophysiology of reno-ureteral colic allows the physician to tailor the best treatment for each patient. PMID- 15133329 TI - Uric acid, phosphate and oxalate stones: treatment and prophylaxis. AB - Medical treatment for the most commonly encountered types of renal stones is described. Nowadays treatment for uric acid stones is well-defined: alkalinizing urine is easy with drugs that are sufficiently active and well enough managed. Relapse is avoided in a high percentage of patients. Medical treatment of phosphate or calcium stones is a more open question as results are far from satisfactory compared with intra- and extra-corporeal approaches which are often minimally invasive and well accepted by both patient and urologist. Relapses are not easy to control because prophylactic measures such as changes in lifestyle and diet are never activated or because they are adopted for a brief period of time. Water therapy is examined, with the choice of water depending on the type of stone, together with drug therapy. Drugs such as citrate, with or without magnesium, and thiazides are considered excellent for curing renal stones and relapses. Although medical therapy has a limited role in many types of stones, its use is decisive in some others. PMID- 15133330 TI - Body weight, diet and water intake in preventing stone disease. AB - Nutrition plays a major role in the pathogenesis of the most widespread forms of nephrolithiasis, i.e. calcium (calcium oxalate and phosphate) and uric acid stone disease. For this reason, dietary measures are the first level of intervention in primary prevention, as well as in secondary prevention of recurrences. An unbalanced diet or particular sensitivity to various foods in stone formers can lead to urinary alterations such as hypercalciuria, hyperoxaluria, hyperuricosuria, hypocitraturia and an excessively acid urinary pH. Over the course of time, these conditions contribute to the formation or recurrence of kidney stones, due to the effect they exert on the lithogenous salt profile. The fundamental aspects of the nutritional approach to the treatment of idiopathic nephrolithiasis are body weight, diet and water intake. This paper will present data resulting from our own investigations and the most significant evidence in literature. PMID- 15133331 TI - Pain killers and antibacterial therapy for kidney colic and stones. AB - In 85% of patients, renal colic is caused by renal-ureteral stones with extrinsic obstructions such as pelvic, retroperitoneal or intestinal abnormalities, and intrinsic reno-ureteral obstructions, e.g. junction pathologies and malformation, accounting for only 10 and 5%, respectively. The objectives of therapy for renal colic therapy are to eliminate pain, preserve renal function and eliminate the obstruction by the excretory pathway. Many drugs can be used to relieve pain: non steroid anti-inflammatory agents (NSAIDs), opioid analgesics, antidiuretic hormone (ADH), loco-regional anesthesia and acupuncture. Opiates are the first choice therapy during pregnancy as no other drug is indicated because of tetragenic potential. Paracetamol (N-acetyl-p-aminophenol) is the only NSAID that is registered for pediatric use because it has none of the adverse side effects that are associated with NSAIDs. Tamsulosin, an alpha-lithic drug, has very recently been included among the drugs that are used for stone expulsion. The rationale underlying its use is that a high concentration of alpha-1D adrenergic receptors has been recently detected in the terminal ureter, especially in the intramural tract. Inhibition of alpha-1D receptor stimulation should relax smooth muscle in the intramural ureteral tract, making stone expulsion easier. PMID- 15133332 TI - Lithiasic obstructive uropathy. Hydronephrosis characterization by magnetic resonance pyelography. AB - OBJECTIVES: The purpose of this study was to evaluate the role of magnetic resonance (MR) pyelography in patients affected by hydronephrosis due to ureteric stones, in order to identify a pyonephrotic condition. MATERIALS AND METHODS: In the last 3 years, 315 patients, who had originally been investigated by ultrasonography, were evaluated with MR pyelography in order to define the etiology of obstruction. In 67 patients hydronephrosis was referred as caused by lithiasis. RESULTS: MR pyelography not only confirmed urinary tract dilatation in all patients, but also identified grade and site of obstruction, both in acute dilatation (25 patients) and in chronic obstructions (42 patients). In 7 patients, MR pyelography documented pyonephrosis that was obviously confirmed by nephrostomic drainage. CONCLUSION: MR pyelography, made with ultrafast breath hold sequences, has a great value in identifying hydronephrosis in patients with ureteric stones. Furthermore, it provides the chance to identify pyonephrosis requiring an immediate drainage of the kidney before major complications develop. PMID- 15133333 TI - Our experience with nifedipine and prednisolone as expulsive therapy for ureteral stones. AB - OBJECTIVES: We evaluated the effect of nifedipine associated with prednisolone in ureteral stone passage. MATERIAL AND METHODS: In our department we enrolled 50 patients with radiopaque ureteral stones. Stone size was 15 mm or less. The patients were divided into two groups: group I included 25 patients who received 30 mg oral treatment of slow-release nifedipine (for a maximum of 20 days) and 25 mg of prednisolone (for a maximum of 10 days) daily. Group II was made up of 25 patients who received 25 mg of prednisolone daily. On request, both groups could use non-steroidal anti-inflammatory drugs. RESULTS: The mean expulsion time was 6 days in group I and 10 days in group II. The average stone size was 12 mm in group I and 12.8 mm in group II. Six patients suspended therapy in group I (5 erythema, 1 stomachache), and seven in group II (3 because of intolerable pain, 4 stomachache). The expulsion success rates were 68% in group I and 81% in group II. CONCLUSIONS: For ureteral stones that do not cause an emergency situation, such as obstructive uropathy, infection or intolerable pain, we suggest expulsive medical treatment with nifedipine and prednisolone, if there are no contraindications to drug use. PMID- 15133335 TI - Dornier Lithotripter S. The first 50 treatments in our department. AB - INTRODUCTION: We assessed the short-term efficacy of extracorporeal shockwave lithotripsy with the Dornier Lithotripter S in the treatment of renal and ureteral stones. MATERIALS AND METHODS: Between February and April 2003, 32 renal and 19 ureteral stones were treated. Patients were evaluated 1 and 3 months afterwards. Stone size and location, total number of shockwaves and the stone free rate were taken into consideration. RESULTS: The stone-free rate for ureteral stones was 63% at 1 month and 84.2% at 3 months. The stone-free rate for renal calculi was 75% at 1 month and 87.5% at 3 months. The overall stone-free rate was 70.6% at 1 month and 86.3% at 3 months. Analgesia was necessary in 12 patients (23.5%). No serious complications were seen, except for one steinstrasse. CONCLUSIONS: The Dornier Lithotripter S is very effective in the treatment of renal and ureteral calculi. PMID- 15133334 TI - ESWL: Comparing two analgesic techniques. Our experience. AB - OBJECTIVE: We compared the effects of intramuscular analgesia versus intramuscular analgesia associated with topical application of Luan (gel containing lidocaine 1%) on the efficacy of extracorporeal shockwave lithotripsy (ESWL). MATERIAL AND METHODS: From January 2001 to November 2002, 300 patients (mean age 46) were treated for kidney or upper ureteral radiopaque stones with ESWL, using a Dornier Compact Delta magneto lithotripter. The patients were divided into two groups: group A, 180 patients, received intramuscular analgesia with Ketorolac 30 mg, Tramadolo 100 mg and, during treatment, intravenous betametasone 4 mg. Group B, 120 patients, received the same pharmacological treatment associated with topical application of Luan. RESULTS: At 3 months, the stone-free rate in group A was 61% compared with 79% in group B. The energy of treatment was less than 13 kV in group A and less than 15 kV in group B. CONCLUSIONS: For ESWL treatment of kidney stones of 20 mm or less and ureteral stones of 15 mm or less, we suggest local analgesia with topical Luan and intramuscular analgesia. This approach increases the success rate of ESWL and reduces the discomfort associated with treatment. PMID- 15133336 TI - Comparing two different ballistic intracorporeal lithotripters in the management of ureteral stones. AB - INTRODUCTION: During the 1990s, two different in situ 'ballistic' lithotripters were introduced into clinical practice. The Swiss Lithoclast (SLC) was the first to be clinically tested. After a few years, a very similar device, the electrokinetic lithotripter (EKL), was described and has recently become available to us. In this paper we compare the clinical efficacy and features of these two ballistic lithotripters in the ureteroscopic treatment of ureteral stones. MATERIALS AND METHODS: Thirty-eight patients with ureteral stones were randomized into two equal groups to undergo fragmentation using the SLC or the EKL via semirigid ureteroscope. In both groups the stones were in the mid- or lower ureter. The following parameters were evaluated: total procedure duration, complete fragmentation rate, time to complete fragmentation, proximal migration rate, complications and need for further procedures. RESULTS: Although not statistically significant, a trend towards a higher fragmentation rate, a shorter time to fragmentation and a slightly higher proximal migration rate emerged in the SLC group. No difference was found in the stone-free rate in the two groups (94.7 vs. 89.4%). Only 2 cases of minor ureteric injury were observed in each group. CONCLUSIONS: As previously described by others, the SLC and EKL are both attractive cost-effective options in the treatment of ureteral stones. Ten years after its introduction, the SLC still remains probably the best choice for most urologists. PMID- 15133337 TI - Expanding applications of the access sheath to ureterolithotripsy of distal ureteral stones. A frustrating experience. AB - OBJECTIVE: Distal ureteral stones are routinely managed with semirigid ureteroscopy without the need for ureteral dilatation, but some conditions, e.g. large stone burden or difficult catheterization of the ureteral meatus, are time consuming and would require a tool to facilitate multiple insertions of the ureteroscope and to avoid ureteral injury. An access sheath is now available and it facilitates ureteroscopy of the upper ureter. To answer the question whether its application could be expanded to the distal ureter is the aim of our paper. PATIENTS AND METHODS: 12 distal ureteral stone patients, 8 with stone size >1 cm and 4 with a large prostate, underwent ballistic ureterolithotripsy with the aid of the Access Sheath. The results of the procedure were compared to a second group of 16 patients from our archives. RESULTS: There was no significant difference in operating time and stone-free rate. Although no complications were related to the ureteroscopy, in 42% (5 patients) of the access group the procedure could be completed only after removing the sheath. CONCLUSION: The Access Sheath does not improve the results of the semirigid ureteroscopy of difficult stones in the distal ureter. PMID- 15133338 TI - Pulmonary function testing in interstitial lung diseases. AB - Interstitial lung diseases (ILDs) are functionally characterized by a restrictive ventilatory defect due to a reduced distensibility of the lung parenchyma. ILD patients also show a reduced exercise tolerance, the main factors limiting exercise capacity being ventilatory and gas exchange abnormalities. Functional abnormalities in ILDs are typical, but not specific. Despite the fact that different lung function patterns have been described among ILDs, they overlap and their practical application to differentiate ILDs is poor. Resting pulmonary function and exercise-induced hypoxemia can aid in defining the prognosis of ILDs and in referring patients for lung transplantation. Additionally, spirometry and diffusing capacity are useful to monitor the response of patients to therapy. PMID- 15133339 TI - Bronchoscopic lung volume reduction: a window for the future? PMID- 15133340 TI - Metalloproteinase-9 in induced sputum correlates with the severity of the late allergen-induced asthmatic response. AB - BACKGROUND: Asthma is characterized by airway inflammation and remodeling in which matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMP) play an important role. Allergen exposure activates the inflammatory/repair process in sensitized subjects. Induced-sputum analysis is a non-invasive method that allows the assessment of changes in inflammatory and remodeling mediators implicated in asthma. OBJECTIVES: To evaluate the changes in MMP-9 and its principal inhibitor (TIMP-1) in sputum and plasma of mild allergic asthmatic subjects after whole-lung allergen challenge. METHODS: Induced sputum and blood samples were obtained at baseline, and 6 and 24 h after challenge. MMP 9 and TIMP-1 levels in sputum and plasma were measured by ELISA. RESULTS: Allergen challenge increased the percentage of sputum eosinophils and MMP-9 levels 6 and 24 h after the challenge compared to baseline levels, but TIMP-1 levels did not vary significantly. A significant correlation was observed between MMP-9 levels at 6 h and the maximum percent fall in FEV(1) during the late response. Throughout the study, MMP-9 levels correlated significantly with the number of neutrophils in sputum. CONCLUSIONS: This study shows that analysis of induced sputum is a useful tool to study the variations in MMP-9 and TIMP-1 levels following allergen challenge, therefore allowing to evaluate their role in allergen-induced airway damage and repair. PMID- 15133341 TI - The position of pranlukast, a cysteinyl leukotriene receptor antagonist, in the long-term treatment of asthma. 5-year follow-up study. AB - BACKGROUND: Adverse effects, tachyphylaxis, and the position of pranlukast, a cysteinyl leukotriene receptor antagonist, in asthma treatment have not been fully established. OBJECTIVES AND METHODS: To address these questions, adverse effects and long-term efficacy of pranlukast were evaluated in 82 patients [28 patients with moderate asthma (group I), 27 with severe persistent asthma not on oral corticosteroid (OCS; group II) and 27 with severe persistent asthma on OCS (group III)] at 4 and 16 weeks. In the following, pranlukast was either withdrawn 1 year after the start of therapy, or if that was not possible due to reappearance of symptoms, the dose of OCS or inhaled corticosteroid (ICS) was reduced. The efficacy of pranlukast was evaluated during 5 years by peak expiratory flow rate (PEFR), and symptom and treatment scores. RESULTS: Adverse reactions appeared in 4 patients (4.9%; diarrhea, dizziness and leg edema). The mean improvement in PEFR on week 16 was 18.5 +/- 2.3, 18.8 +/- 3.2, and 15.2 +/- 3.8% in groups I-III, respectively (p < 0.01, for all groups). However, increases in PEFR in 29 of 72 patients (40.3%) were less than 15%. Pranlukast could not be withdrawn in 28 of 42 responders (66.7%), but their dose of ICS was reduced by 363 +/- 97 microg/day (group II) and that of OCS by 3.4 +/- 0.7 mg/day (group III). Tachyphylaxis was not recognized during the 5-year period. CONCLUSION: Pranlukast is safe when taken for up to 5 years, and is effective irrespective of asthma severity. In the majority of patients with persistent asthma, pranlukast may help to control the disease in the long term. PMID- 15133342 TI - Long-term deterioration of lung function in asthmatic outpatients. AB - BACKGROUND: Although the long-term deterioration of lung function in asthmatic patients has been described, the exact mechanism remains to be determined. OBJECTIVES: The aim of this study was to find correlations between age, sex, atopic status, duration of asthma, asthma severity and the decline in pulmonary function. METHOD: The medical histories of 1,006 randomly chosen asthmatic outpatients were studied and retrospective data on asthma duration, spirometry results, treatment and symptomatology were gathered. A screening spirometry was performed. RESULTS: 598 women and 408 men (age: 44.59, range 12-95 years) participated in the study. Intermittent asthma was diagnosed in 35.4%, chronic mild asthma in 33.4%, moderate asthma in 23.8% and severe asthma in 7.45% of the patients. Statistically significant correlations between patient age, asthma duration and lung function measurements were found. Linear regression revealed the following differences in lung functions per year of asthma duration: FEV1: 0.882% of predicted; FVC: -0.509% of predicted; FEV1/FVC: -0.324% of predicted. The unadjusted annual decline was 80.1 ml/year (p = 0.00003) in FEV1 and 20.5 ml/year (p = 0.036) in FVC. A multiple regression model revealed that asthma severity appears to be the strongest factor influencing pulmonary function (beta = -0.55, p < 0.001 for FEV1). Also, significant associations between pulmonary function measurements, patient age, atopic status and male sex were noted. CONCLUSIONS: The results of this large cohort study show that asthmatic patients develop a progressive decline in pulmonary function correlated with age, sex, duration of asthma and asthma severity. Early diagnosis and intervention is necessary to ameliorate any potential negative impact of asthma on lung function. PMID- 15133343 TI - Decreased serum levels of P-selectin and eosinophil cationic protein in patients with mild asthma after inhaled salbutamol. AB - BACKGROUND: Asthma is a chronic inflammatory disease of the airways associated with selective recruitment of activated eosinophils. P-selectin, a cell adhesion molecule, may be an important controller of the inflammation by mediating selective eosinophil cell influx to the lung. Serum levels of eosinophil cationic protein (ECP) have been used as a marker of eosinophil inflammation, and indirectly as a marker of disease activity of asthma. ECP levels may not be elevated in some patients with asthma, and this fact prompted us to search for additional surrogate markers for monitoring disease activity in asthma. OBJECTIVES: To evaluate whether repeated inhalations of salbutamol, a beta-2 receptor agonist used for bronchodilation, would lead to reduced serum levels of P-selectin and/or ECP. METHODS: Fourteen patients with asymptomatic mild stable asthma were enrolled into a randomised crossover study. Salbutamol was inhaled three times every 3 h. Blood was sampled 4 h after the last inhalation. Nine non treated healthy volunteers served as control subjects. Serum ECP and P-selectin levels were measured using radioimmunoassay and ELISA, respectively. RESULTS: P selectin and ECP levels in serum obtained from asymptomatic asthmatics were close to those of the volunteers, and inter-day variability tended to be lower for levels of P-selectin than for ECP. Significant decreases of P-selectin (p = 0.01) and ECP (p = 0.03) were recorded after salbutamol inhalation. There was no association between the changes in ECP and P-selectin levels in serum. CONCLUSIONS: We conclude that decreases in P-selectin and ECP may have different kinetics, suggesting different pathways of action of salbutamol. We judge that P selectin may be used as a sensitive marker in mild asthma. PMID- 15133344 TI - Occupational therapy and pulmonary rehabilitation of disabled COPD patients. AB - BACKGROUND: Occupational therapy (OT) has been defined as a task of rehabilitation for disabled patients, giving them maximal function and independence to sustain specific activities of daily living. OBJECTIVES: To evaluate the effectiveness of OT as an adjunctive measuring during pulmonary rehabilitation (PR) of hospitalized COPD patients. METHODS: A prospective clinical trial with parallel groups was undertaken in severely disabled COPD patients (n = 71, age 73 +/- 5 years). They were assigned to either OT+PR (n = 47, FEV1 46 +/- 21%pred.) or PR (n = 24, FEV1 44 +/- 12%pred.). PR consisted of eighteen 3-hour daily sessions, whilst OT (domestic activities) was added 3 times a week up to nine 1-hour sessions. Six-min walk (6MWD) with evaluation of BORG dyspnea (D) and leg fatigue (F) scores at end of effort, breathlessness sensation (B) by means of the MRC scale as well as the number of functions lost in the Basic Activity of Daily Living (BADL) categories were assessed as outcomes before (T0) and after (T1) rehabilitation. RESULTS: 6MWD (from 165 +/- 63 to 233 +/- 66 and from 187 +/- 52 to 234 +/- 65 m in the OT+PR and PR groups, respectively), D (from 4.9 +/- 2.1 to 3.2 +/- 1.6 and from 5.3 +/- 2.1 to 3.4 +/- 2.1), F (from 6.1 +/- 0.5 to 4.5 +/- 1.7 and from 5.9 +/- 0.8 to 4.3 +/- 0.8) and B (from 4.3 +/- 0.9 to 3.0 +/- 0.9 and from 4.2 +/- 1.0 to 3.2 +/- 0.8) had similarly improved (p < 0.01) in both groups at T1. The percentage distribution of patients across the BADL categories significantly changed (p = 0.004) in OT+PR (from 17 to 61%, from 70 to 34% and from 23 to 5% in categories A, B and C, respectively) but not in the PR group. CONCLUSIONS: The addition of OT to comprehensive PR is able to specifically improve the outcome of severely disabled COPD inpatients. PMID- 15133345 TI - The effect of continuous positive airway pressure treatment on insulin sensitivity in patients with obstructive sleep apnoea syndrome and type 2 diabetes. AB - BACKGROUND: The obstructive sleep apnoea syndrome (OSA) is a frequent condition, as well as type 2 diabetes mellitus. Both diseases are characterized by insulin resistance. OBJECTIVES: The aim of this study was to establish whether OSA is an independent risk factor for increased insulin resistance in diabetics. For this purpose, we tested the hypothesis that the insulin sensitivity in patients with type 2 diabetes and OSA can be improved by 2 days or 3 months of continuous positive airway pressure (CPAP) treatment. METHODS: In 9 obese patients with type 2 diabetes and OSA [apnoea/hypopnoea index 43.1 +/- 21.3; body mass index (BMI) 37.3 +/- 5.6 kg/m2] and good glycaemic control on oral antidiabetics or on diet alone (HbA1c 6.4 +/- 0.7%), the insulin sensitivity index (ISI) was established by euglycaemic hyperinsulinaemic clamp tests at baseline, after 2 days and after 3 months of effective CPAP treatment. RESULTS: ISI was unchanged after 2 days of CPAP treatment, but was significantly improved after 3 months (4.38 +/- 2.94 vs. 2.74 +/- 2.25 at baseline; p = 0.021), without any significant changes in BMI. Glycaemic control was unaffected after 3 months (HbA1c 6.3 +/- 0.6%; not significant). Fasting leptin levels showed no significant changes. CONCLUSIONS: These results indicate that OSA itself is an independent risk factor for insulin resistance. This effect may be explained by the elevated sympathetic activity in OSA. PMID- 15133346 TI - Usefulness of endobronchial ultrasonography for transbronchial lung biopsies of peripheral lung lesions. AB - BACKGROUND: Peripheral lung lesions are increasing in numbers. Endoscopic diagnosis is essential for the prevention of unnecessary operations. Conventional diagnostic procedures have limitations in availability and results. OBJECTIVES: Endobronchial ultrasonography (EBUS) was investigated as a means to guide transbronchial lung biopsy, to reduce the discomfort during the procedure and to improve diagnostic accuracy. METHODS: In 50 cases, we performed transbronchial lung biopsy combined with EBUS and fluoroscopic guidance. The results were compared to 42 controls assessed by fluoroscopy only. RESULTS: In 38 cases (76%), EBUS could describe the peripheral lesion (33 from inside, including 9 cases with difficulties in fluoroscopic observation, and 5 from an adjacent bronchus, indicating the correct location of the lesion). If successfully placed inside, a change in the patient's position was not required, which helped to reduce patient discomfort. Lung cancer was diagnosed in 24 patients and benign disease in 25 patients; in 1 case diagnosis remained unknown. When the EBUS probe could be introduced inside the lesion, the sensitivity for cancer diagnosis and specificity for cancer exclusion were 100%, respectively (15/15, 18/18). Compared to the controls in whom the biopsy site was determined by fluoroscopy only, the sensitivity tended to be superior by EBUS, although it did not reach statistical significance (p = 0.06). However, specificity and accuracy were statistically significant (both p = 0.02). CONCLUSIONS: When the lesion can be correctly described by EBUS from inside the lesion, EBUS is useful to guide transbronchial lung biopsy, can contribute to a reduction in patient discomfort and improves the accuracy of diagnosis. Additional navigation tools to increase correct positioning of the EBUS probe are desirable. PMID- 15133347 TI - Virtual reality and written assessments are of potential value to determine knowledge and skill in flexible bronchoscopy. AB - BACKGROUND: Technically challenging professions such as those of the defense and transportation industries increasingly use computer-based simulation and written self-learning instruments for education and to determine competency. A structured learning curriculum does not exist, however, for flexible bronchoscopy, a minimally invasive diagnostic procedure performed on thousands of patients by respiratory specialists, otolaryngologists, anesthesiologists, and surgeons worldwide. OBJECTIVE: To explore an analogous strategy of measuring theoretical knowledge in flexible bronchoscopy and specific technical skills using written knowledge assessments and a virtual reality bronchoscopy simulator. METHODS: Twelve trainees from a university pulmonary medicine training program were asked to identify and enter five specific bronchial segments on command using a virtual reality bronchoscopy skill station, and to complete a 50-question examination pertaining to bronchoscopy theory. Their performance scores and opinions pertaining to the use of these methods of assessment were then recorded. RESULTS: Trainees correctly identified and entered 71% of the bronchial segments required on command (median 60%, range 40-100%). Fifty percent (3/6) of the trainees who had performed more than 200 flexible bronchoscopies successfully entered all segments required. None (0/6) of those who had performed less than 200 flexible bronchoscopies correctly located, identified and entered all required segments. Despite disparate performance, all trainees believed that technical skills could be improved through practice and instruction using computer-based simulation. On the written assessment, only 51% of questions were answered correctly (median 52%, range 32-60%). No relationship between technical skill and theoretical knowledge was noted. In addition, neither bronchoscopy skill nor theoretical knowledge were associated with years of training or number of bronchoscopies previously performed. CONCLUSIONS: Trainees concluded that (1) bronchoscopy simulation was realistic, (2) simulator-based practice would help improve technical skills, and (3) a written questionnaire would benefit theoretical knowledge acquisition if designed as a learning instrument. The wide variability noted in this study as well as the lack of a relationship between technical skill, knowledge of bronchoscopy theory, extent of training, and bronchoscopy experience suggest that competency should not be assumed based on years of bronchoscopy training or on an arbitrary number of procedures performed. PMID- 15133348 TI - Pharmacological studies of the effect of wheat grain extract. AB - BACKGROUND: Agricultural farm workers exposed to wheat grain dust are at risk of developing respiratory abnormalities. The pathogenesis of this injury is only partially understood. OBJECTIVES: To determine the effect of wheat grainextract on isolated guinea pig tracheal smooth muscle. METHODS: In the current study, pharmacologic properties of wheat grain extract (WGE) were tested using guinea pig tracheas studied in vitro. Dose-related contractions of nonsensitized guinea pig trachea were demonstrated using these extracts. Pharmacologic studies were performed by pretreating guinea pig tracheal tissue with drugs known to modulate smooth muscle contraction: atropine 10(-6)M, indomethacin 10(-6)M, pyrilamine 10( 6)M, acivicin 10(-5)M, nordihydroguaretic acid (NDGA) 10(-5)M, bromophenacyl bromide (BPB) 10(-5 )M, 3,4,5-trimethoxybenzoic acid-8-(diethylamino)- octyl ester TMB8 10(-5)M, captopril 10(-5)M and capsaicin 5 x 10(-6)M. RESULTS: WGE causes a dose-dependent constriction of guinea pig tracheal smooth muscle. Atropine, pyrilamine, TMB8 and acivicin significantly reduced the contractile effects of the WGE. Inhibition of contraction by blocking of other mediators was significant but less complete. CONCLUSION: We conclude that WGE causes a dose related constriction of airway smooth muscle by nonimmunological mechanisms involving a variety of airway mediators and possibly cholinergic receptors. PMID- 15133349 TI - Positron emission tomography scintigraphy after thoracoscopic talcage. PMID- 15133350 TI - Carcinoma arising in the pleural cavity following pneumonectomy for hydatid disease. AB - We report a case of carcinoma following 42 years of chronic empyema in a patient who underwent surgery for a hydatid cyst at the age of 3. At the time of diagnosis, an esophageal fistula was observed and treated with cyanoacrylate. We hypothesize that chronic inflammation of the pleura, caused by decades of empyema, associated with the presence of heterotopic squamous epithelium due to a long-standing esophago-pleural fistula, led to neoplastic transformation. PMID- 15133351 TI - Rhabdomyolysis after the administration of itraconazole to an asthmatic patient with bronchopulmonary aspergillosis. AB - It is known that the coadministration of itraconazole with cholesterol-lowering statins may induce muscle damage. We describe the case of a patient with steroid dependent asthma, steroid-induced severe myopathy and allergic bronchopulmonary aspergillosis who developed rhabdomyolysis and acute renal failure following a approximately 6-week treatment with itraconazole, even without the concomitant use of HMG-CoA reductase inhibitors. PMID- 15133352 TI - Sarcoidosis presenting as metastatic bony disease. A case report and review of the literature on vertebral body sarcoidosis. AB - Although asymptomatic lytic bony lesions of the phalanges of the hands and feet are not uncommon in patients with sarcoidosis, involvement of the vertebral bodies is rare. Because these lesions can mimic other diseases of the bones on radionuclide scans and magnetic resonance imaging (MRI), this case emphasizes the importance of obtaining tissue and excluding malignancy and infection before the diagnosis can be established with confidence PMID- 15133353 TI - Pulmonary masses presenting 11 years after abdominal surgery. PMID- 15133354 TI - Pleural effusion in yellow nail syndrome: treatment with bilateral pleuro peritoneal shunts. PMID- 15133355 TI - Efficacy of a microwave antenna for ablation of the tricuspid valve--inferior vena cava isthmus in dogs as a treatment for type 1 atrial flutter. AB - INTRODUCTION: Radiofrequency catheter ablation of the tricuspid valve-inferior vena cava (TV-IVC) isthmus for treatment of atrial flutter (AFL), may in some cases require a large number of energy applications and a long procedure and fluoroscopy time. AIMS OF STUDY: Therefore, we studied the safety and efficacy of a 4 cm long microwave antenna mounted on a steerable 9Fr catheter for linear ablation of the TV-IVC isthmus. METHODS: In 6 anesthetized dogs, multi-electrode catheters were positioned in the coronary sinus (decapolar), at the His bundle (quadripolar) and around the TV annulus (decapolar) for pacing and recording atrial activation sequences before and after ablation. The microwave antenna was then positioned across the TV-IVC isthmus from the TV annulus (identified by equal A and V potentials) to the inferior vena cava with slight traction on the catheter to ensure adequate endocardial contact. Microwave energy was then applied at a fixed power for 120 seconds during each ablation attempt. Ablation was repeated until bi-directional isthmus block was demonstrated during pacing from the coronary sinus ostium and low lateral right atrium, respectively. RESULTS: Linear microwave ablation of the TV-IVC isthmus was completed in all ten dogs using a total of 2.6 +/- 1.17 energy applications per dog. Power was applied in a range of 45-50 watts. There were no acute procedural complications. Bi directional TV-IVC isthmus block was achieved in all ten dogs, as demonstrated by a strictly descending activation wavefront in the ipsilateral atrial wall, during pacing from the CSO and LLRA respectively. In addition, after ablation conduction time to the LLRA during pacing from the CSO increased from 52 +/- 16.62 before to 87 +/- 12.74 msec (p <.05), and to the CSO during pacing from the LLRA from 51 +/ 12.43 before to 79.50 +/- 9.85 msec (p <.05). Gross and histological examination of the TV-IVC isthmus after ablation revealed continuous transmural lesions, ranging from 3-5 mm in width, spanning the entire TV-IVC isthmus in all ten dogs. CONCLUSIONS: (1) Microwave ablation of the TV-IVC isthmus was safe and effective in this study. (2) Ablation of the entire width and thickness of the TV-IVC isthmus can be rapidly achieved using a long microwave antenna in a fixed trans isthmus position. PMID- 15133356 TI - Transmembraneous irrigation of multipolar radiofrequency ablation catheters: induction of linear lesions encircling the pulmonary vein ostium without the risk of coagulum formation? AB - INTRODUCTION: Pulmonary vein (PV) isolation for the curative treatment of atrial fibrillation using conventional radiofrequency ablation (RF) catheters with the point by point technique is time consuming and carries a remaining risk for thrombembolic complications. AIMS OF THE STUDY: Aim of the present in vivo study was to evaluate feasibility and safety of a novel multipolar irrigated ablation catheter designed to create contiguous lesions encircling the PV ostium in a single ablation position. METHODS: The entire ablation section (tripolar, length of each electrode 22 mm, interelectrode distance 2 mm, helix radius: 9 and 10 mm) of the 7F RF catheter (Encirclr, Medtronic, MN, USA) was covered by a porous membrane (pore size 30 micron) providing continuous irrigation. The helical formed catheter was used in two different experimental settings. Initially, a thigh muscle preparation has been performed in 7 anesthetized sheep in order to evaluate the development of lesions at different power level (40-80 W) and RF duration (30-90 sec). The ablation catheter was placed at the surface of the thigh muscle in a perpendicular position (0.1 N contact pressure) and perfused with heparinized blood (250 ml/min, 37C degrees ). Irrigation was provided with a flow rate of 10 ml/min. The resulted lesion morphology was evaluated with regard to coagulum or crater formation and lesion depth and diameter. Subsequently in 9 anesthetized sheep intracardiac ablation has been achieved with 50 W and an irrigation flow of 10 ml/min. Transseptal puncture and RF ablations were guided using fluoroscopy and intracardiac echocardiography (ICE, Acuson, USA). Endpoint of the intracardiac RF applications was the reduction of local electrogram amplitude >50%. RF applications were achieved at both atrial appendages and in the orifices of the coronary sinus (CS), the vena cava inferior (VCI) and PV. Following RF ablation all animals were sacrificed and following in vivo staining (2% TTC) macroscopically and histologically investigations of the lesions were performed. RESULTS: At the thigh muscle preparation 57 RF applications have been performed. The lesion depth was homogeneous without gaps between the ablation electrodes. There was a significant increase comparing 30 with 90 sec of RF duration for 40, 50 and 60 W applications respectively: 40 W: 1.1 +/- 0.4 vs. 3.6 +/- 0.5; 50 W: 1.2 +/- 0.3 vs. 4.6 +/- 0.4 mm and 60 W: 2.6 +/- 0.6 vs. 4.8 +/- 0.5 mm. All applications with 80 W (n = 3) had to be terminated due to immediate increase of impedance >150 omega. Late impedance rises (>60 sec) without occurrence of coagulum formation have been observed in 1 out of 4 RF applications with 60 W.A total of 85 RF applications could be achieved intracardiacally in the right atrium (right atrial appendage n = 18, ostium of the coronary sinus n = 12, ostium of the inferior caval vein: n = 12) and in the left atrium (left atrial appendage: n = 15, ostium of the PV: n = 28). ICE guided positioning of the catheter and showed during all applications no coagulum formation at the electrode or impedance rise (>150 Omega). Reduction of local electrograms (>50%) were observed following 48 out 85 (56%) RF applications. The lesions showed a homogeneous depth of 4 +/- 2 mm and a width 5 +/- 2 mm at the surface. No charring or crater formation could be observed in any of the lesions. CONCLUSIONS: In the present in vivo studies it could be demonstrated that long irrigated ablation electrodes induce continuous lesions without the risk of thrombus formation at the electrode. Increase of RF duration from 30 to 90 seconds with power setting of 40-60 W, respectively, created deeper lesions without the risk of thrombus formation. Thus, the helical formed irrigated ablation catheter appears to be appropriate for simplified PV isolation. PMID- 15133357 TI - Implantable loop recorders: a novel method to judge patient perception of atrial fibrillation. Preliminary results from a pilot study. AB - INTRODUCTION: At the present time, several techniques are used or are under investigation for atrial fibrillation (AF) therapy. Nowadays, no well-defined target for such therapies has been yet completely identified. Furthermore, AF is an arrhythmia with high rates of recurrences, both symptomatic and asymptotic. Thus the measure of therapy success rates not only based on symptom perception remains a goal to be reached. AIMS OF THE STUDY: This study investigates the role of an implantable loop recorder (ILR) as an additional tool to identify initiating and perpetuating mechanisms of AF. The role of right atrial linear ablation (RALA) procedures is also investigated using the monitoring capabilities of the ILR. METHODS AND RESULTS: Nine patients (mean age 63.8 +/- 5.9) with paroxysmal AF were referred to our institution as candidates for AF ablation. All patients (pts) had in their medical history several years of AF episodes. Therefore pts were aware of AF related symptoms. Six of them were implanted with an ILR before ablation and were monitored one month before and six months after the procedure. The ILRs stored 54 patient activated events (PAE) and 124 automatically activated events (AAE). 68% of PAEs and 67% of AAEs were classified as appropriate. Most common reasons for inappropriate detections were premature atrial or ventricular contractions among PAEs and undersensing among AAEs. The arrhythmia onset was properly identified in 4 pts (44%). The average AF recurrence rate was 10.8 +/- 3.5 ep/month before ablation and 5.0 +/- 1.8 ep/month after the procedure ( p = 0.042). CONCLUSION: The ILR may be a helpful tool in monitoring pts undergoing ablation. Dedicated AF detection characteristics could give additional value to the device. RALA appears as a feasible, safe and relatively effective first approach in AF therapy. PMID- 15133358 TI - Predictors of early and late recurrence of atrial fibrillation after catheter ablation of paroxysmal atrial fibrillation. AB - INTRODUCTION: The outcome of patients with early recurrence of atrial fibrillation (AF) (within one month) after ablation procedure is controversial. Furthermore, the predictors of early and late (up to mean follow-up 30 months) recurrence of AF are not investigated in depth. AIMS OF THE STUDY: The aim of the present study was to investigate the predictors of early and late recurrence of AF after catheter ablation of arrhythmogenic foci initiating AF in patients with paroxysmal AF. METHODS AND RESULTS: The study included 207 patients (155 men; mean age 62 +/- 13 years) who received catheter ablation of paroxysmal AF. Eighty one (39%) patients had early recurrence of AF. Five clinical variables were related to the early recurrence of AF: (1) old age (>/=65 years) ( P = 0.004); (2) presence of associated cardiovascular disease ( P = 0.01); (3) presence of multiple AF foci ( P = 0.004); (4) presence of AF foci from left atrial free wall ( P = 0.039); (5) left atrial enlargement ( P = 0.038). Multivariate analysis demonstrated that presence of multiple AF foci could predict early recurrence of AF ( P = 0.013; ratio = 2.24; 95% CI 1.18 to 4.25). During the follow-up period (30 +/- 11 months), 70 (34%) patients had late recurrence of AF, and two clinical variables were related to the late recurrence of AF: (1) presence of early recurrence of AF ( P = 0.025); (2) presence of multiple AF foci ( P = 0.034). Multivariate analysis found that presence of early recurrence of AF could predict late recurrence of AF ( P = 0.046; hazard ratio = 1.62; 95% CI 1.01 to 2.59). Late recurrence of AF happened in 35 (43%) of the 81 patients with early recurrence of AF, and in 35 (28%) of the 126 patients without early recurrence of AF. CONCLUSIONS: Early AF recurrence could predict late AF recurrence. PMID- 15133359 TI - A randomized comparison of alternative techniques to achieve coronary sinus cannulation during biventricular implantation procedures. AB - INTRODUCTION: Biventricular pacing system implantation is a time-consuming and challenging procedure. A critical step in biventricular pacemaker implantation is coronary sinus (CS) cannulation. CS cannulation can be achieved either using dedicated guiding catheters (guiding catheter alone positioning strategy, GCA) or with the aid of an electrophysiology catheter advanced inside the guiding catheter (electrophysiology catheter aided positioning strategy, EPA). AIM OF THE STUDY: To evaluate whether the EPA technique is useful for reducing CS cannulation time compared to a conventional GCA technique. METHODS: Thirty-four consecutive patients were randomly assigned to the GCA (18 patients) or EPA (16 patients) CS cannulation strategy. RESULTS: Time to successful catheterization of CS was 5.0 +/- 2.4 min in the EPA group versus 10.1 +/- 5.4 min in the GCA group p = 0.004. Fluoroscopy time was 4.6 +/- 2.3 min in the EPA group versus 9.2 +/- 4.9 min in the GCA group p = 0.004. Total contrast dye volume to search and engage the CS ostium was 0.0 ml in the EPA group versus 14.3 +/- 3.4 ml in the GCA group p < 0.001. CONCLUSIONS: Cannulation of CS with the adjunct of an electrophysiology catheter to dedicated delivery systems significantly reduces procedural time, fluoroscopy time and contrast dye volume compared to a conventional strategy. PMID- 15133360 TI - Preventricular far-field sensing in the atrial channel of dual chamber pacemakers -an occasional cause of inappropriate mode switch. AB - INTRODUCTION AND AIMS OF THE STUDY: Atrial oversensing may trigger false positive mode switch to an asynchronous mode in dual chamber pacemakers. While myopotential oversensing and far-field R wave sensing within the postventricular atrial refractory period are well characterized, data about oversensing (near field P wave or far-field R wave) within the atrioventricular delay is limited. Aim of the study was to determine the incidence of preventricular oversensing in the atrial channel of current dual chamber pacemakers. METHODS: Consecutive patients with dual chamber pacemakers who were in sinus rhythm and who showed no myopotential oversensing were included in the study. Atrial sensitivity was programmed to the maximal available value and atrioventricular delay was prolonged when necessary for intrinsic atrioventricular conduction. RESULTS: Ten out of 100 (10%) patients showed oversensing within the atrioventricular delay at highest levels of atrial sensitivity, median sensing threshold for the signals was 0.35 mV, and the median coupling interval between preventricular atrial oversensing and the ventricular sensed event was 20 ms. CONCLUSION: The incidence of inappropriate mode switch caused by atrial preventricular oversensing related to near-field P wave or far-field R wave oversensing in dual chamber pacemakers is up to 10%. Interpretation of pacemaker Holters should consider oversensing within the atrioventricular delay as a differential diagnosis to true episodes of atrial tachyarrhythmias. PMID- 15133361 TI - Management of inadvertent left ventricular permanent pacing. AB - Inadvertent implantation of a pacemaker lead in the left ventricle is an uncommon complication. We report a case of a permanent pacemaker lead inadvertently placed through the left subclavian artery, across the aortic valve into the left ventricle. A chest X-ray one month after the procedure showed an unusual course of the lead and a 12-lead ECG and a transthoracic echocardiogram confirmed the diagnosis. The patient refused surgical removal and remained on full anticoagulation. No clinical events were recorded during a 3-year follow-up. In such cases we propose life-long full anticoagulation as an alternative to surgical lead extraction. PMID- 15133362 TI - Electrophysiological characterization of cardiac veins in humans. AB - BACKGROUND: The coronary sinus is a complex structure with a surrounding myocardial coat and muscle bundles that course within it. The purpose of this study was to evaluate the electrical activity of the coronary sinus (CS), great cardiac vein (GCV) and related structures, such as the Vein of Marshall (VOM). METHODS AND RESULTS: Data obtained from adult ( n = 114) and pediatric patients ( n = 16) were analyzed. The width of atrial electrograms (EGMs) within the CS at a basic pacing cycle length of 600 ms was 46 +/- 7.4 ms (mean +/- SD) vs. 29.7 +/- 6.3 ms in the GCV ( p < 0.01). With decremental pacing the width of the EGM within the CS at 300 ms increased to 66.6 +/- 8.5 ms ( p < 0.1 compared to CS EGM at pacing cycle length of 600 ms). The width of the EGM within the GCV increased from 29.7 +/- 6.3 ms at a pacing cycle length of 600 ms to 34.6 +/- 6.0 at 300 ms ( p = NS). There were no significant differences in the atrial EGM width between CS and GCV in the pediatric patients. CONCLUSIONS: We conclude that atrial electrograms are wider in the CS but not in the GCV. This finding can be explained by the presence of a myocardial coat around the CS. The rate response characteristics of the atrial electrograms within the CS are consistent with a lack of tight coupling between muscle bundles and the CS musculature. Further, the absence of such differences in pediatric patients could partly explain relative differences in types of supraventricular arrhythmias seen in different age groups. PMID- 15133363 TI - Atrioventricular nodal reentrant tachycardia with multiple discontinuities in the atrioventricular node conduction curve: immediate success rates of radiofrequency ablation and long-term clinical follow-up results as compared to patients with single or no AH-jumps. AB - BACKGROUND: Some patients with atrioventricular nodal reentrant tachycardia (AVNRT) demonstrate multiple discontinuities (AH jump) in their antegrade AV node conduction curves. We evaluated and compared the immediate success rates, procedure-related complications, long-term clinical follow-up results and recurrence rates after slow pathway ablation in patients with multiple versus single or no AH jumps. METHODS: The study group consists of 278 consecutive patients (mean age 36.6 +/- 15.7) who underwent ablation for typical AVNRT, divided into three categories according to the number of AH jumps (>/=50 ms) before ablation: Group-1 consisted of 63 patients (23%) with continuous AV node function curves; Group-2 of 183 patients (66%) with a single jump and Group-3 of 32 (12%) patients showing more than one AH jumps. RESULTS: Age was significantly higher in Group-3 as compared to Group-1 (43 +/- 18 years vs. 34 +/- 16 years, p = 0.020). The electrophysiological features of AVNRT did not differ among groups. Before ablation, the maximum AH interval was significantly longer in Group-3 as compared to Groups-1 and -2 ( p < 0.001 for both). AV node antegrade ERP was significantly shorter in Group-3 than in Group-2, both before and after ablation ( p < 0.050 for both). AV node Wenckebach cycle length (WCL) was shorter in Group 3 as compared to both Groups-1 and -2, before and after ablation ( p < 0.050 for all). AV node WCL was prolonged significantly in all groups after ablation ( p < 0.001 for all). Residual dual pathways were present in 37 of 278 patients (13%) after ablation and were significantly more frequent in Group-3 than Group-2 (31% vs. 15%, p = 0.023). CONCLUSIONS: Patients with multiple AH jumps are older and more often have residual dual atrioventricular nodal pathway physiology after successful ablation but these features do not affect the immediate and long-term success rates of slow pathway ablation as compared to patients with single or no AH jumps. PMID- 15133364 TI - Bundle branch reentrant tachycardia in a patient with normal ventricular function. AB - We report an unusual case of bundle branch reentrant tachycardia, in a patient with normal left ventricular function, cured by radiofrequency catheter ablation. However, the long-term prognosis of these patients is uncertain. We discuss the indications for an implantable defibrillator in this group. PMID- 15133365 TI - An unusual case of right ventricular outflow tract tachycardia. AB - In this report, we describe an unusual case of right ventricular outflow tract (RVOT) tachycardia with episodes of repetitive monomorphic ventricular tachycardia (VT), paroxysmal sustained VT and incessant monomorphic VT of the same morphology. Diltiazem, adenosine, or metoprolol failed to interrupt these arrhythmias. However, administration of intravenous propafenone completely eliminated all ventricular ectopic activity. Electrophysiologic study performed off propafenone showed that the ventricular ectopic activity originated from a single locus at the anterior wall of the RVOT. Two radiofrequency applications at this site resulted in complete elimination of ventricular ectopic activity. PMID- 15133366 TI - Effect of heart rate and isoproterenol on pulmonary vein flow velocity following radiofrequency ablation: a Doppler color flow imaging study. AB - INTRODUCTION: Application of radiofrequency energy at pulmonary vein (PV) ostium during focal atrial fibrillation (AF) ablation procedures increases flow velocity due to PV narrowing. Factors unrelated to ablation that effect PV flow velocity have not been described. AIMS OF THE STUDY: The purpose of this study was to evaluate, using intracardiac echocardiography (ICE) imaging, the effect of isoproterenol (ISO) and heart rate (HR) on PV flow velocity Pre- and Post ablation. METHODS AND RESULTS: In 31 patients with AF undergoing LA-PV ostial ablation involving at least one PV ostium, an ICE catheter was placed in the RA to image and detect PV flow. PV ostial peak velocity was assessed in sinus rhythm Pre-, Post-ablation, during and after ISO (up to 20 microg/min). To separate HR versus ISO effect, PV velocity was measured during atrial pacing (after HR returned to baseline) at pacing rate matching HR with ISO. PV ostial velocity was assessed with ISO and pacing in 30 non-ablated and 33 ablated PVs. Ostial velocities of non-ablated PVs during ISO infusion (117 +/- 42 cm/s) were greater ( p < 0.03) than those during atrial pacing (78 +/- 26 cm/s) at matched HR (116 +/- 20, range 92-150 bpm). Ostial PV flow velocities of ablated PVs increased from 59 +/- 17 (30-95) cm/s Pre- to 95 +/- 25 (58-136) cm/s Post-ablation. During ISO infusion PV flow velocities in ablated PVs (118 +/- 34 cm/s) were also greater ( p < 0.03) than those during atrial pacing (96 +/- 37 cm/s) at matched HR (116 +/- 14, range 92-130 bpm). Atrial pacing alone produced no significant difference in PV flow velocities measured Pre- or Postablation. CONCLUSION: ISO appears to increase ostial flow velocity of ablated and non-ablated PVs independent of HR effect. These effects are important to recognize when PV velocity is used as an index for interpreting the impact of PV ostial lesions on functionally significant PV narrowing. PMID- 15133368 TI - Nonsurgical transthoracic epicardial approach in patients with ventricular tachycardia and previous cardiac surgery. AB - INTRODUCTION: The subxyphoid pericardial mapping approach can be used to facilitate catheter ablation of postmyocardial-infarction ventricular tachycardia (post-MI VT), but the presence of dense adhesions is thought to preclude this approach in patients who have previously undergone open-chest cardiac surgery. AIMS OF THE STUDY: This study reports the first use of a nonsurgical transthoracic epicardial approach in patients with scar-related VT and previous cardiac surgery. METHODS: Five patients with a mean age of 67 +/- 10 years, left ventricular ejection fraction (LVEF) of 40 +/- 4.3%) and recurrent VT occurring 7 months to 10 years after cardiac surgery underwent combined endocardial and epicardial mapping and ablation during the same session. Because pericardial adhesions were anticipated to be denser in the anterior wall, the nonsurgical transthoracic epicardial puncture was directed to the inferior wall of the left ventricle. Failure to interrupt VT with radio frequency (RF) energy pulses delivered at the best endocardial or epicardial site prompted changing from one approach to the other. RESULTS: During the epicardial puncture procedure, the contrast medium accumulated in the inferior wall instead of spreading around the cardiac silhouette. The pericardial sac could be entered in all patients, and mapping of the inferolateral epicardial wall of the left ventricle was feasible. Fourteen VTs were induced, of which 8 could not be mapped because of poor hemodynamic tolerance. Three of the remaining 6 mappable VTs were eliminated by endocardial ablation, 2 required an epicardial RF pulse to be rendered noninducible, and 1 VT was not eliminated. No intra- or postprocedural complications were noted despite full heparinization. CONCLUSION: Nonsurgical transthoracic epicardial catheter mapping and ablation of epicardial VT related to the inferolateral left ventricular wall are feasible in patients who have previously undergone open- cardiac surgery. PMID- 15133367 TI - Endothelial damage and activation of the hemostatic system during radiofrequency catheter isolation of pulmonary veins. AB - AIMS: To determine the systemic thrombogenic effect of radiofrequency catheter isolation of the pulmonary veins (PVI) in the treatment of atrial fibrillation. METHODS AND RESULTS: We studied endothelial damage marker (von Willebrand factor [vWf]), fibrinolysis markers (tissue plasminogen activator [t-PA], plasminogen activator inhibitor-1 [PAI-1]) and coagulation activation markers (D-dimer [DD]) in 30 patients (pts) undergoing PVI. Heparin was administered continuously after double transseptal puncture in all pts. Concentrations of vWf and t-PA were significantly increased after accomplishing PVI compared to the baseline values, and elevated levels persisted 24 hours later ( p < 0.01). PAI-1 levels decreased following PVI compared to the baseline levels ( p = 0.02). PAI-1 levels normalized 24 hours after the procedure. DD increased continuously during the procedure with the peak following PVI ( p < 0.01). Higher DD concentrations persisted 24 hours later ( p = 0.02). In a multivariate analysis, total procedure time correlated significantly with the peak vWf and DD concentrations, while total RF energy dose correlated only with peak vWf ( r = 0.82). Time to heparin administration correlated with DD levels prior to the first RF pulse ( r = 0.83, p < 0.01) as well as after PVI ( r = 0.75, p < 0.01). A group of patients heparinized within the first hour of the PVI procedure had normal preablation DD levels and significantly mitigated DD levels following PVI compared to the group of patients heparinized later ( p < 0.01). CONCLUSIONS: Pulmonary vein ablations cause an increased systemic procoagulant state as reflected by fibrin turnover, fibrinolysis activation and endothelial perturbation. The activation of the coagulation cascade could be decreased by early heparin administration. PMID- 15133369 TI - Spontaneous transition from atrial fibrillation to typical atrial flutter during catheter ablation of the pulmonary vein. AB - A 71-year-old male patient was admitted for catheter ablation of the pulmonary veins to treat paroxysmal atrial fibrillation. Atrial fibrillation originating from the left superior pulmonary vein was induced after a pause of atrial pacing under isoproterenol infusion and became sustained. Spontaneous transition from atrial fibrillation to typical atrial flutter was noted after complete isolation of the pulmonary vein focus from the left atrium. Subsequently linear ablation of the cavotricuspid isthmus was created with completely bi-directional isthmus conduction block. We hypothesized that ectopic pulmonary vein focus played an important role in the spontaneous conversion of atrial fibrillation to typical atrial flutter, and complete isolation of the pulmonary vein could stop the spontaneous transition between the two atrial tachyarrhythmias. PMID- 15133370 TI - Bernard Lown. PMID- 15133372 TI - Coronary risk factor management in the framework of a community hospital-based ambulatory exercise training program. AB - Ischemic heart disease is a chronic illness that causes major mortality and morbidity. Angiographic studies have shown the effectiveness of exercise programs, in combination with aggressive lipid management, in reversing or slowing the progression of atherosclerotic coronary disease. Despite these studies, participation in supervised programs that combine exercise and risk factor management is limited. The authors measured the ability of a community hospital-based ambulatory cardiac rehabilitation program to recruit patients and to facilitate reduction of risk factors that have been demonstrated to influence progression of disease. Patients were recruited from a single community hospital for an ambulatory exercise training and cardiac risk-factor management program, and clinical and laboratory data was collected periodically. Recruited patients participated in a minimum 3-month period of training and counseling by a multidisciplinary team with follow-up measurements of weight, lipid profile, blood pressure, and exercise capacity. Thirty-two percent of the eligible hospitalized patients were successfully recruited into the program. Dropout rates over the initial 3 months were low (25%). Improvement in low-density lipoprotein cholesterol level (-4.5%), high-density lipoprotein cholesterol level (+7%), body mass index (-2%), systolic blood pressure (-3%), and maximum metabolic equivalents (+25%) were comparable to levels achieved in studies showing angiographic stabilization and/or regression of disease. Implementation of a community hospital-based risk management exercise program is an effective method for improving the long-term management of patients with chronic ischemic heart disease. PMID- 15133373 TI - Relaxation response for Veterans Affairs patients with congestive heart failure: results from a qualitative study within a clinical trial. AB - Fifty-seven veterans with congestive heart failure were interviewed about their experiences and changes after participating in a three-armed randomized trial: relaxation response (RR) training, cardiac education, and usual care. The interviews were tape-recorded, transcribed, and analyzed. Half of the 20 RR group interviewees reported physical improvements, and 13 reported emotional improvements. These improvements went beyond disease management to lifestyle changes and improved family/friends relationships. Five of 16 cardiac education group interviewees reported physical improvements, and eight reported emotional improvements. These improvements consisted of a better understanding of the disease and resulted in feeling more at ease. None of the usual care group interviewees reported any improvement from study participation. Although group support contributed to the benefits reported by RR and cardiac education groups, the use of the RR techniques seems to be the factor that distinguished the improvements. The value of the RR in congestive heart failure health care is suggested by the results. PMID- 15133374 TI - Should the National Cholesterol Education Program guidelines be changed for persons at high risk for cardiovascular events? PMID- 15133375 TI - Cardiovascular actions of selective estrogen receptor modulators and phytoestrogens. AB - Cardiovascular disease is the leading cause of death among men and women in Western societies. Over the past decade, interest in a better understanding of gender differences in cardiovascular disease has heightened. Concomitantly, the use of hormone therapy for cardiovascular risk reduction in postmenopausal women has come into question in light of recent landmark clinical studies casting doubt on the benefits of this therapy. As a consequence, alternatives to conventional hormone replacement, including selective estrogen receptor modulators and phytoestrogens, have attracted considerable attention. The authors provide an up to-date review of the clinical actions of selective estrogen receptor modulators on cardiovascular disease. The actions of tamoxifen, raloxifene, droloxifene, and soy phytoestrogens are discussed in the context of cardiovascular disease epidemiology, coronary events, clinical markers of cardiovascular risk, and vascular function. In addition, the authors' current understanding of the mechanism of action of these agents is discussed and recommendations for clinical practice are reviewed. PMID- 15133376 TI - Depression and acute myocardial infarction. AB - A number of studies have demonstrated a relationship between depression and low perceived social support and increased cardiac morbidity and mortality in patients with coronary heart disease. There is also evidence that depression increases the risk of acute myocardial infarction and morbidity and mortality following it. This review examines those studies that have investigated these relationships as well as those that have attempted to explain them on the basis of various pathophysiologic mechanisms. Among the latter are studies that have shown that selective serotonin reuptake inhibitors are beneficial in the treatment of depression and that they appear to reverse the enhanced platelet activity observed in depressed patients with acute myocardial infarction. Depression increases hospital length of stay, procedures, readmission rates, and the cost of medical care. Much remains to be elucidated concerning the roles of depression and low perceived social support in predisposing to acute myocardial infarction and to increased morbidity and mortality following it. However, sufficient scientific evidence exists for physicians to make efforts to diagnose and treat depression to reduce the concurrent risk of acute myocardial infarction and morbidity and mortality following it. PMID- 15133377 TI - VISP and INVEST. PMID- 15133378 TI - Acculturation and cardiovascular disease risk in midlife immigrant women from the former Soviet Union. AB - This study examines relationships among acculturation, depression, and cardiovascular risk factors in midlife women from the former Soviet Union and identifies factors predicting Framingham Risk Scores. Data were collected at baseline and 1 year later from 218 participants in a longitudinal study of postimmigration health. The leading risk factors were obesity, dyslipidemia, and depression. Older women had lower American Behavioral Acculturation subscale scores, higher Russian Behavioral Acculturation subscale scores, and higher depression scores. Length of residence was significantly correlated with American behavioral acculturation but not Russian behavioral acculturation. Baseline body mass index, both acculturation scores, and depression scores predicted Framingham Risk Scores after 1 year, but serum glucose did not. The results suggest that contrary to findings in other immigrant groups, women from the former Soviet Union may decrease their risk for coronary heart disease as they assume a more American lifestyle. Nursing interventions to address the high cardiovascular disease risk in this population are suggested. PMID- 15133379 TI - The impact of daily sleep duration on health: a review of the literature. AB - A healthy amount of sleep is paramount to leading a healthy and productive lifestyle. Although chronic sleep loss is common in today's society, many people are unaware of the potential adverse health effects of habitual sleep restriction. Under strict experimental conditions, short-term restriction of sleep results in a variety of adverse physiologic effects, including hypertension, activation of the sympathetic nervous system, impairment of glucose control, and increased inflammation. A variety of epidemiologic studies have also suggested an association between self-reported sleep duration and long-term health. Individuals who report both an increased (>8 h/d) or reduced (<7 h/d) sleep duration are at modestly increased risk of all-cause mortality, cardiovascular disease, and developing symptomatic diabetes. Although the data are not definitive, these studies suggest that sleep should not be considered a luxury, but an important component of a healthful lifestyle. PMID- 15133380 TI - Cardiovascular magnetic resonance imaging. AB - Recent advancements in magnetic resonance imaging hardware and software permit the assessment of cardiovascular structure and function at rest and during exercise or pharmacology-induced cardiac stress. With these developments, knowledge of cardiovascular imaging protocols in the magnetic resonance imaging environment is critical for nursing personnel. The purpose of this article is to review information pertinent to working in a magnetic resonance imaging environment and to describe the requirements of nursing personnel performing cardiovascular magnetic resonance imaging examinations. PMID- 15133381 TI - Now I lay me down to sleep--but how long is long enough? PMID- 15133382 TI - Cardiovascular nursing in Austria. PMID- 15133383 TI - The Portfolio Diet to reduce the risk of heart disease. PMID- 15133384 TI - Phosphodiesterase inhibitors in the management of primary pulmonary hypertension. PMID- 15133385 TI - Get with the guidelines for acute myocardial infarction: a core measure improvement process. PMID- 15133386 TI - Conduction delay in heart failure patients? PMID- 15133387 TI - After the antibiotics and breast cancer study: what to tell patients. PMID- 15133390 TI - Which symbol should be used to represent medicine? PMID- 15133391 TI - Ramsay Hunt syndrome revisited. PMID- 15133392 TI - Ultraviolet carcinogenesis in nonmelanoma skin cancer part II: review and update on epidemiologic correlations. AB - The relationship between ultraviolet radiation and nonmelanoma skin cancer (NMSC) is further elucidated by a review of case-control studies relating type of exposure to the development of NMSC. Intermittent sun exposure is important in the pathogenesis of basal cell carcinoma, whereas cumulative exposure is important for both basal cell carcinoma and squamous cell carcinoma. The regional distribution of NMSC is also in areas of sun exposure. Furthermore, there are inherent risk factors for the development of NMSC that include hair and eye colors. Once an individual develops NMSC, he/she has increased risk of developing basal cell carcinoma, squamous cell carcinoma, or melanoma. PMID- 15133393 TI - Alpha-hydroxy acids: unapproved uses or indications. AB - Alpha-hydroxy acids have been used for rejuvenation since ancient times, and now there are several on the market. Depending on the concentration, some have been shown to be effective as peeling agents and for rejuvenation. Glycolic acid and lactic acid are the alpha-hydroxy acids most frequently used in cosmetics, although there are many others used in combination. Some of the most striking advances in dermatology have followed the off-label use of drugs, which is widespread and unavoidable, but a well founded scientific approach to an individual patient's pathology must be emphasized as the number of products, regimens, and adjuncts increases exponentially in the cosmetic field. In this article the authors review some unapproved uses of alpha-hydroxy acids. More published data on the scientific value of these off-label indications that proves whether they are effective or not is needed. PMID- 15133394 TI - HIV revisited: the global impact of the HIV/AIDS epidemic. AB - This review is intended as an update on modern trends in the global impact and epidemiology of human immunodeficiency virus infection for physicians who are not acquired immunodeficiency syndrome experts. Africa has been the most affected, but epidemics are spreading in Asia and Russia. Therefore, physicians should be informed about seroconversion disease and human immunodeficiency virus diagnosis as well as the impact of sexually transmitted infections on many stages of human immunodeficiency virus. International treatment guidelines are available. Highly active antiretroviral therapy has been the mainstream therapy since 1996, but all drugs--regardless of class used--have potent side effects, many of which are dermatologic. Others affect the neurologic, hematopoietic, cerebral, and abdominal systems, and drug interactions are common. Lipodystrophy is a common, long-term side effect that is still not well understood. Broader use of highly active antiretroviral therapy has highlighted viral resistance. This is reviewed, and a simple explanation of therapeutic monitoring is provided. PMID- 15133395 TI - Humira (adalimumab). PMID- 15133396 TI - Cosmetic science regulation. PMID- 15133397 TI - Hyaluronic acid in office practice. PMID- 15133398 TI - Pseudofolliculitis barbae. PMID- 15133399 TI - Lepra vaccine. PMID- 15133400 TI - Auricular granulomatosis. PMID- 15133401 TI - Agiokeratomas of the vulva: possible association with radiotherapy. PMID- 15133402 TI - Mosaic hair regrowth pattern of ophiasis and androgenic alopecia in a patient with alopecia areata totalis. PMID- 15133403 TI - Effective treatment of hypertension without medication: is it possible? PMID- 15133404 TI - Hypertension treatment in the ambulatory setting: comparison by race and gender in a national survey. AB - To determine if different therapies are used in different racial groups and by gender, data from the National Ambulatory Medical Care Survey and the National Hospital Ambulatory Care Survey (national probability samples of outpatient visits) were used. All visits for hypertension in 1999 and 2000 were reviewed. Survey weights were applied to obtain national estimates. Provision of therapies by gender and race/ethnicity (white, African American, Hispanic, Asian) was examined. Over 137 million visits for hypertension care were made during 1999 and 2000. Diet and exercise counseling were performed at a low percentage of visits (35% and 26% of visits, respectively). The most common antihypertensive agent prescribed was angiotensin-converting enzyme inhibitors or angiotensin-receptor blockers (28%), while first-line drugs, diuretics (23%) and b blockers (15%), which are recommended by national committees, were prescribed less frequently. Asians and Hispanics were more likely to receive counseling on diet (Asians: odds ratio [OR], 2.29; 95% confidence interval [CI], 1.45-3.60; Hispanics: OR, 2.51; 95% CI, 1.18-5.33) and exercise (Asians: OR, 2.44; 95% CI, 1.35-4.42; Hispanics: OR, 3.28; 95% CI, 1.50-7.21) than non-Hispanic whites. African Americans were more likely to be prescribed calcium channel blockers (OR, 1.51; 95% CI, 1.20 1.91) and diuretics (OR, 1.37; 95% CI, 1.08-1.74). Low use of recommended therapies was found. Although variation by race was seen, it did not systematically favor groups associated with poor outcomes. PMID- 15133405 TI - The antihypertensive efficacy and safety of a chronotherapeutic formulation of propranolol in patients with hypertension. AB - This study evaluated the antihypertensive efficacy and tolerability of a chronotherapeutic formulation of propranolol designed for nighttime dosing (propranolol controlled release [CR]). A total of 434 patients with mild-to moderate hypertension were randomized to placebo or to one of four doses of propranolol CR (80, 120, 160, or 640 mg/d). At baseline, the mean morning blood pressures were similar in each treatment group and averaged 152/101 mm Hg. After 8 weeks of treatment, morning diastolic blood pressure, the primary efficacy measurement, was significantly reduced from baseline in placebo (-6.98 mm Hg) and all propranolol groups (p<0.001). The decreases ranged from 10.1 mm Hg in the 80 mg/d group to 11.0 mm Hg in the 120-mg/d group and were significantly larger than placebo in the 120-, 160-, and 640-mg/d groups (p<0.05). Blood pressure measured in the evening (trough) demonstrated similar antihypertensive efficacy. Heart rate and rate-pressure product were reduced in a dose-related manner by propranolol CR. The formulation was well tolerated with only fatigue and dizziness being reported more frequently than in the placebo group. Propranolol CR is an effective antihypertensive formulation that may reduce blood pressure during the morning period of maximum cardiovascular risk. PMID- 15133406 TI - The natural treatment of hypertension. AB - The goal of this review is to evaluate the efficacy of commonly available dietary supplements in the treatment of hypertension, using the average blood pressure reduction achieved with the implementation of lifestyle modifications as a standard. For this reason, the authors focus on the antihypertensive potential of these agents rather than pharmacology, pharmacokinetics, adverse effects, or supplement-drug interactions. For the purpose of this review, dietary supplements are defined as exhibiting some evidence of benefit if a systolic blood pressure reduction of 9.0 mm Hg or greater and/or a diastolic blood pressure reduction of 5.0 mm Hg or greater has been observed in previously published, peer-reviewed trials. These defining limits are based on the average blood pressure reduction associated with the implementation of certain lifestyle modifications. Agents with some evidence of benefit include coenzyme Q10, fish oil, garlic, vitamin C, and L-arginine. PMID- 15133407 TI - Hypertension and the elderly: more than just blood pressure control. AB - Hypertension is a major risk factor for cardiovascular disease in both young and elderly persons; therefore, good blood pressure control is at the center of improved cardiovascular health. The recently issued seventh report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure and the European Society of Hypertension/European Society of Cardiology 2003 guidelines for hypertension management emphasize the importance of treatment efficacy rather than age in treating elderly persons with hypertension. Most hypertension clinical trials have been carried out with younger hypertensives, but this is changing with trials such as the Systolic Hypertension in the Elderly Program, the first Swedish Trial of Old Patients With Hypertension, and the Systolic Hypertension in Europe trial. These trials have clearly demonstrated the benefits of good blood pressure control in reducing the risk of stroke in elderly persons. With many safe and effective antihypertensive drugs on the market, the question becomes how elderly persons should be treated. Elderly patients often have isolated systolic hypertension, which is related to loss of arterial elasticity or compliance with aging and is more recalcitrant to treatment than essential hypertension. In addition, with advancing age there is the likelihood that other disease states are present in addition to hypertension. The newer antihypertensive drugs that interfere with the renin angiotensin system, such as angiotensin-converting enzyme inhibitors and angiotensin-receptor blockers, have the potential of improving cardiovascular outcomes in elderly persons in addition to offering effective blood pressure reduction. Their use should be considered within a comprehensive risk assessment that includes individualized risk-benefit considerations. PMID- 15133408 TI - Effect of drug therapy for heart failure on quality of life. AB - Symptomatic heart failure interferes with a patient's quality of life (QOL) by limiting his or her ability to perform physical tasks and daily activities and by lowering his or her sense of psychological well-being. Therefore, in addition to decreasing mortality and morbidity, improving QOL should be an important goal when selecting pharmacotherapy. QOL questionnaires, both generic and disease specific, are used widely, but in randomized controlled trials of heart failure treatments, QOL has not been a routine study end point. Beta blockers and angiotensin-converting enzyme inhibitors, medications widely used in the management of heart failure and hypertension--one of the most common causes of heart failure--have been associated with negative, neutral, and modestly positive QOL effects. Angiotensin receptor blockers, combined with other therapy, including angiotensin-converting enzyme inhibitors (usually with a diuretic) and/or b blockers in heart failure, have produced improvements in QOL. Patients with hypertension whose blood pressure has been lowered have also experienced an improvement in QOL scores. With current heart failure regimens prolonging life, improving QOL becomes an even more essential end point in assessing the effectiveness of new medications, whether used alone or in combination with standard therapy. PMID- 15133409 TI - Clinical problems in the management of hypertension 1) Prehypertension: should we treat? 2)The very elderly: how should we treat. PMID- 15133410 TI - Treatment of octogenarians: should we and how. PMID- 15133412 TI - Hypertension in Hispanics. PMID- 15133413 TI - Minoxidil: an underused vasodilator for resistant or severe hypertension. AB - Minoxidil is a direct vasodilator introduced in the early 1970s for the treatment of hypertension. It is capable of reducing blood pressure in most persons with resistant hypertension where therapy has failed with multidrug regimens. Minoxidil's effect can be limited because of an increase in pulse rate and/or sodium (and water) retention. The latter may prove quite debilitating in some patients. Thus, minoxidil is generally administered with both a diuretic and an agent that can keep pulse rate in check, such as a beta blocker or a combined alpha-beta blocker. The prominent tachycardia with minoxidil can aggravate myocardial ischemia and, if long-standing, leads to left ventricular hypertrophy. Minoxidil has a particularly annoying side effect of hypertrichosis that may limit its use, particularly among women. Minoxidil use is infrequently associated with the idiosyncratic onset of a pericardial effusion. If a patient's hypertension is severe enough to warrant minoxidil therapy, a hypertension specialist should probably become involved in the patient's care. The use of this medication should be limited in view of the availability of effective agents with fewer side effects. There is, however, a place for minoxidil in the treatment of resistant hypertension especially in patients with advanced renal disease. PMID- 15133415 TI - Impact of aging on hostility in coronary patients and effects of cardiac rehabilitation and exercise training in elderly persons. AB - Hostility is a coronary artery disease (CAD) risk factor affecting recovery after major CAD events. Yet, few data exist on the impact of aging on hostility in CAD patients or the effects of cardiac rehabilitation on elderly patients with high hostility. The authors studied 500 consecutive patients following CAD events to determine the impact of aging on hostility scores, the prevalence of hostility symptoms, and the response of elderly persons with hostility symptoms to cardiac rehabilitation programs. Overall hostility scores (p<0.01) and prevalence of hostility symptoms (8% in 268 elderly vs. 28% in 81 younger patients [aged <50 years]) were inversely related to age (p<0.01). Elderly persons with hostility symptoms had greater weight (p=0.02), four times higher anxiety and depression scores (p<0.0001), two times higher scores for somatization (p<0.0001), and 17% lower scores for quality of life (p<0.001) compared with elderly persons without hostility symptoms. Marked reduction in hostility and improvements in other risk factors occurred following rehabilitation. These results demonstrate the inverse relationship between age and hostility symptoms in patients with CAD. Although hostility symptoms are relatively uncommon in elderly persons with CAD, these patients have more adverse CAD risk profiles and still have marked improvements following cardiac rehabilitation and exercise training programs. PMID- 15133416 TI - Treatment of elderly hypertensive patients with a delayed-release verapamil formulation in a community-based trial. AB - This report of a practice-based clinical trial describes an open-label, multicenter dose-titration study of an elderly (age >or=65 years) subset of patients (N=628) with systolic blood pressures between 140 and 179 mm Hg or diastolic blood pressures between 90 and 109 mm Hg, to assess the effects of the Chronotherapeutic Oral Drug Absorption System (CODAS) formulation of verapamil hydrochloride. After starting 200 mg/d at bedtime, dosing was titrated to a maximum of 400 mg/d at 4-week intervals to achieve a target blood pressure of <140/<90 mm Hg using morning blood pressure measurements. Target blood pressure was reached in 57.1% of the elderly patients with CODAS verapamil monotherapy. A diastolic response (<90 mm Hg or a 10 mm Hg reduction from baseline) was achieved in 89.5% of these subjects, and a systolic blood pressure response (<140 mm Hg or 10% reduction from baseline) was attained in 75.5%. The percentages of patients achieving target blood pressure or the diastolic and systolic blood pressure responses were comparable to those previously reported for younger patients. It is notable that although 359 of the 628 patients reached control on treatment, 182 of the remaining 269 noncontrolled patients were not titrated to higher doses, indicating that even with a well tolerated drug there may be reluctance among clinicians to increase doses. CODAS verapamil was found to be efficacious and well tolerated among elderly hypertensive patients in this community trial. PMID- 15133417 TI - Subclinical cardiovascular disease in older adults: insights from the Cardiovascular Health Study. AB - Knowledge about the epidemiology of subclinical cardiovascular disease (SCVD) in older adults may hold the key for improved opportunities for primary prevention of cardiovascular disease (CVD), a top clinical and public health priority. This review reports findings on the prevalence of SCVD and the ability of SCVD measures to predict incident and adverse outcomes from one of the largest (N=5888) and most comprehensive prospective observational studies on SCVD in older adults, the Cardiovascular Health Study. According to a composite index that combined SCVD measures from different vascular beds, the overall prevalence of SCVD was 37%, making it as common as clinically overt CVD in older adults. SCVD measures strongly predicted incident CVD, stroke, mortality, frailty, and physical and cognitive decline, even after adjustment for traditional CVD risk factors. Ongoing research will address the potential use of SCVD for clinical decision making in older adults. PMID- 15133418 TI - Lifestyles of older adults: can we influence cardiovascular risk in older adults? AB - Influences of lifestyle habits on cardiovascular disease risk among older adults are not well established. The authors present evidence from the Cardiovascular Health Study that dietary, physical activity, and smoking habits assessed late in life are associated with cardiovascular disease risk among adults aged 65 years or older. Persons consuming fatty fish twice per week had a 47% lower risk of coronary death compared with those who consumed fatty fish less than once per month, while cereal fiber intake (about two whole-grain bread slices per day) was associated with a 14% lower risk of myocardial infarction or stroke. Modest alcohol intake (1-6 drinks per week) predicted the fewest subclinical cerebrovascular abnormalities. Compared with little activity, moderate and high leisure-time activity predicted 28% and 44% lower mortality, respectively, while compared with nonexercisers, low, moderate, and high exercise intensity predicted 30%, 37%, and 53% more years of healthy life, respectively. Former and current smokers had 25% and 44% fewer years of healthy life than those who never smoked; lifetime smoking (pack-years) predicted higher mortality. Clinical practice and public health implications, gaps in knowledge, and future research directions are summarized. PMID- 15133419 TI - Medications and cardiovascular health in older adults: room for improvement in prevention and treatment. PMID- 15133420 TI - Ethical issues in the management of geriatric cardiac patients. PMID- 15133422 TI - Assessment and treatment of lipids in elderly persons. PMID- 15133423 TI - Dyslipidemia as a risk factor at elderly age. AB - Although the relative risk of high total cholesterol or low high-density lipoprotein cholesterol decreases with advancing age, this is offset by the greater absolute and attributable risk in an elderly population. Abnormal lipid levels are associated with increased coronary risk in an elderly population, and both increased total and decreased high-density lipoprotein cholesterol levels predict coronary death at elderly age. About one third of elderly men and one half of elderly women have cholesterol levels >240 mg/dL. Remediable dyslipidemia is highly relevant at elderly age and should guide recommendations for cardiovascular disease prevention. PMID- 15133424 TI - Statin therapy in the elderly: observational and randomized controlled trials support event reduction. AB - 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitor (statin) therapy for older patients at high risk of cardiovascular events is supported by data from randomized clinical trials and observational data, including the Cardiovascular Health Study, an LDS Hospital/University of Utah cohort study, and a study from New York Medical College. Randomized trials such as the Scandinavian Simvastatin Survival Study, the Cholesterol and Recurrent Events trial, the Heart Protection Study, and the Prospective Pravastatin Pooling Project included large numbers of elderly patients and uniformly support safety and event reduction in these groups. Although debate continues over the appropriateness of treating elderly persons and the strength of data to support a decision to treat, it is clear that older patients have the highest absolute risk of events and the potential to show the greatest benefit. PMID- 15133425 TI - A prospective study of pravastatin in the elderly at risk: new hope for older persons. AB - The role of cholesterol in coronary heart disease (CHD) and stroke in elderly persons is controversial. Statin studies have generally included mostly middle aged men, been targeted at either primary or secondary prevention, and lasted 5-6 years. In contrast, the PROspective Study of Pravastatin in the Elderly at Risk was a double-blind, randomized, placebo-controlled primary and secondary prevention trial that examined the effects of pravastatin 40 mg/d on the risk of CHD, cerebral vascular events, and cognitive function over 3 years in 5804 high risk elderly persons (mean age 75 years; 52% women). Pravastatin therapy significantly improved lipid levels: low-density lipoprotein cholesterol (-34%), high-density lipoprotein cholesterol (+5%), and triglycerides (-13%) and produced no adverse effects on liver function tests or myopathy. Moreover, pravastatin reduced the primary end point (CHD death, nonfatal myocardial infarction, and stroke) by 15% (p=0.014), including 24% reduction in CHD death (p=0.043) and 19% reduction in combined CHD death and nonfatal myocardial infarction (p=0.006). Although transient ischemic attacks fell by 25% (p=0.05) with pravastatin, there were no significant reductions in stroke or improvements in cognitive function. These results demonstrate the benefits of pravastatin in primary and, particularly, secondary prevention in elderly populations and further support the benefits of statin therapy in high-risk elderly persons. PMID- 15133426 TI - Lipid and statin effects on stroke and dementia. AB - Stroke and dementia are among the leading causes of morbidity and mortality in the elderly. Although elevated cholesterol level has not been a significant risk factor for cerebrovascular disease events, statins produce pleiotropic effects beyond cholesterol lowering and have been shown to reduce major cerebrovascular events, including stroke and transient ischemic attack. This review looks at the effects of this class of agents on major diseases of the central nervous system. PMID- 15133427 TI - Treatment of hyperlipidemia in elderly persons with exercise training, nonpharmacologic therapy, and drug combinations. AB - Hyperlipidemia is a major modifiable risk factor for the primary and secondary prevention of cardiovascular diseases in the elderly population. Although statin therapy remains the mainstay of treatment for hyperlipidemia in the elderly, other therapies, including exercise training, plant stanols and sterols, soluble fiber, and drug combinations (especially ezetimibe, niacin, and/or fibrates alone or combined with statins) are also effective and important lipid therapies for the elderly population. PMID- 15133428 TI - Statin safety: what to know. PMID- 15133429 TI - [The history of intestinal suturing: it's decisive influence on digestive surgery]. PMID- 15133430 TI - [Treatment of hepatocellular carcinoma in the cirrhotic liver]. AB - The incidence of hepatocellular carcinoma (HCC) in cirrhotic patients is increasing. Despite advances in imaging and laboratory screening which allow earlier diagnosis, the surgeon is all too often confronted with an HCC of advanced stage or arising in the setting of severe cirrhosis; this severely limits the treatment possibilities. Treatment options are constrained not only by the characteristics of the tumor but also by hepatocellular reserve, severity of portal hypertension, and the general condition of the host. "Curative treatments" envisage the complete eradication of the malignancy; they include liver transplantation, resection, or tumor destruction by radiofrequency or alcohol ablation. They are most effective in the early stages of HCC. Total hepatectomy and transplantation, by far the most complex surgical therapy, also has the best results avoiding the all-too-frequent local recurrence of HCC in the residual liver. Other medical and interventional treatments (chemo-embolization, radiotherapy with lipiodol) can only slow the progress of the HCC. Goals for the future include more precise and directed screening of the population at risk, and better chemopreventive and chemotherapeutic treatments. PMID- 15133431 TI - [Esophageal diverticula]. AB - Esophageal diverticula are classified by location-phrenoesophageal (Zenker's diverticulum-70%), thoracic and mediastinal (10%), and epiphrenic (20%). Almost all esophageal diverticula are acquired pulsion diverticula. The most common symptoms are dysphagia, regurgitation, thoracic pain, and pulmonary manifestations related to aspiration. Barium swallow and upper endoscopy will help to establish the diagnosis while esophageal manometry may reveal underlying dysmotility. Diverticula should not be treated unless they are symptomatic. The treatment of Zenker's diverticulum is surgical and consists of either diverticulectomy or diverticular suspension with a myotomy of the cricopharyngeus muscle via cervical approach. Transoral endoscopic stapled diverticulostomy is a new and simple approach which may become the treatment of choice, particularly in elderly and high-risk patients. Treatment of diverticula of the mid and low esophagus must take into account any motor anomalies or associated lesions. Diverticulectomy with esophageal myotomy and an anti-reflux procedure through a left thoracotomy is the standard approach, but endoscopic approaches seem feasible, particularly for epiphrenic diverticula, and may become the norm in years to come. PMID- 15133432 TI - [Laparoscopic colectomy involving the hepatic flexure]. PMID- 15133433 TI - [Surgical treatment of bleeding duodenal ulcer by antroduodenal resection, exclusion of the ulcer base, and duodenal closure after partial intramural dissection?]. PMID- 15133434 TI - [How to get into the "frozen" abdomen]. PMID- 15133435 TI - [Splenic metastasis from a primary carcinoma of the piriform sinus]. PMID- 15133436 TI - [Surgical management of retrorectal tumors]. AB - Retrorectal tumors (RRT) constitute an anatomical grouping of various tumors of different nature, both benign and malignant. The diversity of their presentation, surgical management, and prognosis are illustrated by five clinical cases. A simple categorization would distinguish vestigial tumors (whether cystic or solid), congenital nonvestigial tumors such as chordoma, and tumors of neural or bony origin. Imaging by CT scan and by MRI will usually determine the nature of the tumor and its relationship to the surrounding anatomical structures. The principle of treatment is complete removal with free margins. The surgical approach may be posterior, anterior or combined depending on the nature and the size of the lesion and on how high it is situated relative to the second sacral vertebra. Complete resection may be both difficult and bloody. Sacral segments may need to be resected either for reasons of surgical approach or to obtain clear margins. Rectal resection is rarely necessary. The prognosis of these lesions depends on the nature of the tumor and particularly on the quality and completeness of the resection. PMID- 15133437 TI - [Cancer of the gastric cardia with a pulmonary nodule]. PMID- 15133438 TI - [Surgical behavior: ethical problems arising from anger, aggravation, fatigue, ageing, and inappropriate sexual relationships]. PMID- 15133439 TI - [Congress of the American College of Surgeons (Chicago, October 19-23, 2003)]. PMID- 15133440 TI - [Chlamydial mesenteric abscess with small bowel obstruction]. PMID- 15133442 TI - [Surgery for malignant pleural mesothelioma]. AB - Malignant pleural mesothelioma (MPM) is an aggressive cancer of the pleura that is usually caused by exposure to asbestos. The incidence of MPM has risen for some decades and is expected to peak between 2010 and 2020. Current surgical treatment involves in a multimodality regimen with radiation and multiple-drug chemotherapy. All currently proposed therapeutic strategies are in total agreement with the international Mesothelioma Interest Group TNM staging system. Schematically: for stage Ia (early stage disease), the therapeutic approach is generally neo-adjuvant intrapleural treatment using cytikines followed by surgical pleurectomy; for more advanced disease (stage Ib, II and III), a multimodal treatment combining extra-pleural pneumonectomy, radiotherapy and multiple-drug chemotherapy, including in all cases cisplatin, is proposed. Recently, results using this multiple modality approach have been favorable especially for patients with epithelial histology, negative resection margins and no metastases to extrapleural lymph nodes; for stage IV (unresectable tumor), palliative treatment is indicated. Early results have been encouraging and the use of recent drugs should allow more optimal treatment. PMID- 15133443 TI - [Thoracic aspergillosis: indications for surgery for a multifaceted disease!]. AB - We reviewed the different clinical forms of thoracic aspergillosis and detailed surgical options. Classical aspergiloma where a tuft of Aspergillus grows in a parenchymal cavity is the most well-known entity. Simple forms (little clinical expression, thin-walled cavity without impact on neighboring tIssue) can be distinguished from complex forms (poor general status, thickened cavity, sequellae). Surgery is the last resort for complex forms, but the procedure is benign for simple forms allowing interruption of the spontaneous evolution. Pleural aspergillosis is a common complication of the excision procedure, whether performed early or at mid-term. Thoracoplasty is often required due to the Volume of parenchyma removed. Surgery can be proposed for acute invasive aspergillosis in two situations: to prevent cataclysmic hemoptysis due to a paravascular lesion, or for resection of sequestered mycotic deposits which could lead to generalized reinfection. Semi-invasive aspergillosis is usually observed in areas of post-radiation fibrosis where the typical aspergillar excavation appears after the initial phase of invasion leading to lobular pneumonia. Thoracoplasty is often the only surgical option. Ulcerated aspergillar tracheobronchitis is observed after (heart)-lung transplantation and raises the risk of characteristic invasive aspergillosis. Finally rare observations of parietal aspergillosis have been treated by surgical resection in combination with systemic antifungal agents. Multidisciplinary consultation is required to establish the most appropriate approach. PMID- 15133444 TI - [Lung transplantation]. AB - Lung transplantation is indicated for patients with cystic fibrosis, emphysema, pulmonary fibrosis or pulmonary hypertension whose life expectancy is less than two years. Criteria of severity are detailed. Three types of transplantation can be proposed: single lung transplant for fibrosis and dry emphysema; bilateral lung transplant for cystic fibrosis, and certain types of emphysema and pulmonary hypertension; heart-lung transplant for pulmonary hypertension and Eisenmenger syndrome. Due to insufficient supply of donor organs, one quarter of the candidates die on the waiting list and the limit for inscription is often 60 years. Postoperative mortality at two months is about 15% and is related to graft dysfunction, infection, bronchial complications,... Acute rejection usually occurs during the first year. Chronic rejection is expressed by obliterating bronchiolitis, the leading cause of death after one year. There is a risk of cancer (EBV-induced lymphoproliferative syndromes and skin cancer). Five-year survival is still only about 50%. Immunosuppressor treatments still cause numerous adverse effects (hypertension, renal toxicity...); function and quality of-life have however greatly improved. PMID- 15133445 TI - [Pneumothorax surgery]. AB - Surgery remains the mainstay treatment of recurrent pneumothorax. We recall the therapeutic modalities and indications of surgery in case of pneumothorax. We then compare postoperative outcome, recurrence rate and chronic pain with regard to techniques and surgical approach. Current video-thoracoscopic or axillary thoracotomy procedures for bullous disease with pleurodesis allow a low rate of morbidity and recurrence after primary or secondary spontaneous pneumothorax. PMID- 15133446 TI - [Thoracic sympathectomy: treatment for hyperhidrosis]. AB - Hyperhidrosis is a benign functional anomaly which is highly stressful for the patient. Active management is required. Several medical options are available but are often ineffective. The thoracic sympathic system plays a fundamental role in propagating stimulation of sudoral gland secretion. Endoscopic thoracic sympatecomy thus provides a radical treatment for severe palmar and axillary hyperhidrosis. We describe the technique used in our unit and present results and possible complications. This method has been used by many teams for several Years and despite some differences, most confirm major patient benefit. Phenomena of transferred sudation are frequent by are usually not invalidating. Patients should however be informed of this possibility because the effect is often irreversible. PMID- 15133447 TI - [Surgery for chylothorax]. AB - Chyle is lymph fluid of intestinal origin containing fat digestion products. Chylothorax is produced by leaks from the thoracic duct or from one of its collaterals subsequent to valve incompetence. These leaks may be due to trauma (post-surgical chylothorax, the most frequent) or to spontaneous rupture of a lymph vessel distended by chyle reflux, the thoracic duct itself being pathological. When the thoracic duct is interrupted (obstruction, agenesis), chylothorax may occur from leakage due to reflux within substitution collateral pathways diverting the flow of chyle into the venoux confluents of the neck. Medical treatment is always attempted first: evacuation of chylothorax by drainage and fat-free diet or parenteral nutrition. Recently, treatment with octreotide has been found to be beneficial. Surgery consists in thoracic duct ligation or suture of leaking collaterals. In difficult cases, when the chyle leakage cannot be identified, pleurodesis is the only option. PMID- 15133448 TI - [Surgical management of emphysema]. AB - Surgery is an important therapeutic option for emphysema patients with invalidating dyspnea and poor quality-of-life. Preoperative tests must determine the degree of functional impairment (dyspnea score, walking test, quality-of life) and evaluate lesion reversibility (imaging, function tests, TLCO, blood gases, scintigraphy, right microcatheterism) and assess the patient's general health status. Besides lung transplantation, the only surgical alternative is resection which, depending on the type of parenchymal damage, can involve excision of bullae or volume reduction. Several modalities can be proposed: atypical resection of the apexes via sternotomy, multiple unilateral atypical resection, simple lobectomy. The choice depends on the distribution of the parenchymal destruction and also on the severity of the emphysema and the patient's age. Operative mortality is now well below 10%. Volume reduction provides significant functional improvement in 80% of patients but with a temporary effect (4-5 years). Bullae excision is particularly important since functional recovery is achieved early and persists. PMID- 15133449 TI - [Surgery for diaphragmatic palsy]. AB - Diaphragmatic palsy leads to a permanent ascension of one or both hemi-diaphragms with highly variable functional impact. The underlying mechanisms can be divided into two main categories: neurological or muscular disorder leading to peripheral dysfunction; defective or non-transmitted central command causing central dysfunction. A complete morphological and functional work-up is required to determine the circumstances leading to diaphragmatic palsy and the uni- or bilateral nature of the paralysis. The entire phreno-diaphragmatic transmission chain from the cranium to the diaphragmatic muscle must be analyzed to search for a local cause. Function tests are used to examine central command and transmission, function of the phrenic nerve, and the capacity of the diaphragmatic muscle to generate sufficient pressure for efficacious ventilation. Once indirect causes of diaphragmatic ascension (independent of the phreno diaphragmatic system) have been ruled out, surgery may be proposed for symptomatic, permanent and irreversible diaphragmatic paralysis. A tension procedure may be sufficient in the event of eventration with or without phrenic palsy. For well-selected patients with central paralysis due to supraspinal lesions with intact nerves and muscles, implantation of a phrenic pacemaker may be helpful to eliminate positive pressure mechanical ventilation and restore more physiological respiration. PMID- 15133450 TI - [Surgical treatment of post-embolism pulmonary hypertension]. AB - Pulmonary hypertension is a serous condition which, after a long history as an orphan disease, has raised renewed interest due to the development of efficacious therapeutic options including lung transplantation and continuous infusion of prostacycline. Bilateral endarteriectomy of the pulmonary arteries is another possibility for post-embolism pulmonary hypertension. The procedure is complex and must be performed in conditions of cardiac arrest and deep hypothermia but, unlike transplantation, provides definitive cure. Recognizing the post-embolic nature of pulmonary hypertension is not simple because old episodes of venous thrombosis or embolus migration are not found in 50% of patients. Segmentary defects on the perfusion scintigraphy contrasting with the homogeneous respiratory scintigraphy is the primary diagnostic feature. Lesions must be located in a main trunk or at the origin of lobular or segmentary branches to be accessible to endarteriectomy. An antero-posterior and lateral angiogram of each lung and a multiple-array helicoidal angioscan performed with a precise protocol by an experienced team are needed to identify the localization of the lesions. If the pulmonary resistance determined at right catheterism is correlated with anatomic obstruction, the risk of mortality of pulmonary endarteriectomy is low, offering patients a significant chance for normal or nearly normal cardiorespiratory function. PMID- 15133451 TI - Cytokine induced apoptosis in human retinoblastoma cells. AB - PURPOSE: To determine potential anti-proliferative properties of interferon-gamma (IFN-gamma) and tumor necrosis factor (TNF-alpha) on human retinoblastoma cells. METHODS: Fluorescent antibody staining was used to detect IFN-gamma and TNF-alpha receptors on the cells. Y79 and Weri Rb-1 cells were exposed to IFN-gamma alone, TNF-alpha alone, or a combination of IFN-gamma and TNF-alpha, and apoptosis was measured by caspase 3 activation and annexin V staining. Cell cycle arrest was measured by BrdU incorporation and FACS analysis. RESULTS: Both cell lines expressed receptors for IFN-gamma and TNF-alpha. There appeared to be two populations of both receptors in the Weri Rb-1 cell line. Apoptosis was induced in Y79 cells by IFN-gamma, but not TNF-alpha, and the combination of the cytokines did not increase apoptosis above IFN-gamma alone in Y79 cells. Apoptosis was induced in Weri Rb-1 cells only upon exposure to both cytokines. The cell cycle was not significantly altered in either cell line. CONCLUSIONS: Human retinoblastoma cells respond to IFN-gamma or a combination of IFN-gamma and TNF-alpha by becoming apoptotic, but Y79 and Weri Rb-1 cells behave differently. The differential response of the two cell lines is not due to a lack of expression of IFN-gamma or TNF-alpha receptors. The data raise the possibility that differences in apoptotic pathways exist between the two cell lines with interesting implications for the induction of apoptosis as a therapy for retinoblastoma. PMID- 15133452 TI - Moving beyond imagination. PMID- 15133453 TI - Developing an emergency department based Special Operations Team: Robert Wood Johnson University Hospital's experience. AB - Robert Wood Johnson University Hospital initiated an emergency department based Special Operations Team as a way to help prepare staff for the care of hazardous material incidents (HAZMAT) victims and the unexpected consequences of a mass casualty incident. The team evolved over a period of 5 years and is now able to provide significant educational offerings, policy and procedure review and participation in extensive planning efforts in the hospital and community. This article will review the process and present future goals of the team. PMID- 15133454 TI - Earthquakes in El Salvador: a descriptive study of health concerns in a rural community and the clinical implications: Part III--Mental health and psychosocial effects. AB - BACKGROUND: In 2001, the mountain town of San Sebastian, El Salvador experienced a series of earthquakes that affected the livelihood of its people. METHODS: A convenience sample of 100 households of 594 inhabitants of San Sebastian and the surrounding rural farming areas was completed. One study participant for each household was evaluated for mental health and psychosocial changes after the earthquakes. The participant's questionnaire was used to investigate the relationship between physical health, access to health care, housing, food and water, and the occurrence of negative mental health markers six months after the disasters. RESULTS: Findings indicate that the majority (67%) of respondents experienced 6 or more mental health complaints. Risk factors associated with multiple negative mental health symptoms included change in household income and loss of job, a new illness or a new injury in the household, reliance on healthcare services since the earthquake, and managing a chronic illness. CONCLUSION: Findings indicate a need for rapid, acute mental health screening with at-risk groups and the need to educate the entire community regarding what medical and mental health treatments are available to reduce barriers to treatment and increase public awareness. PMID- 15133455 TI - The San Diego County wildfires: perspectives of healthcare providers [corrected]. AB - The wildfires of October 2003 burned a total of 10% of the county of San Diego, California. Poor air quality contributed to an increased number of patients seeking emergency services, including healthcare providers affected by smoke and ash in hospital ventilation systems. Two large hospitals with special patient populations were threatened by rapidly approaching fires and had to plan for total evacuations in a very short time frame. A number of medical professionals were forced to prioritize responding to the hospital's call for increased staff during the disaster and the need to evacuate their own homes. PMID- 15133456 TI - The Rhode Island Medical Emergency Distribution System (MEDS). AB - The State of Rhode Island conducted an exercise to obtain and dispense a large volume of emergency medical supplies in response to a mass casualty incident. The exercise was conducted in stages that included requesting supplies from the Strategic National Stockpile and distributing the supplies around the state. The lessons learned included how to better structure an exercise, what types of problems were encountered with requesting and distributing supplies, how to better work with members of the private medical community who are not involved in disaster planning, and how to become aware of the needs of special population groups. PMID- 15133461 TI - Safety of vaccines affected by a power outage. PMID- 15133467 TI - Optimum treatment of severe sepsis and septic shock: evidence in support of the recommendations. AB - Severe sepsis and septic shock are among the most common causes of death in noncoronary intensive care units. The incidence of sepsis has been increasing over the past two decades, and is predicted to continue to rise over the next 20 years. While our understanding of the complex pathophysiologic alterations that occur in severe sepsis and septic shock has increased greatly asa result of recent clinical and preclinical studies, mortality associated with the disorder remains unacceptably high. Despite these new insights, the cornerstone of therapy continues to be early recognition, prompt initiation of effective antibiotic therapy, and source control, and goal-directed hemodynamic, ventilatory,and metabolic support as necessary. To date, attempts to reduce mortality with innovative, predominantly anti-inflammatory therapeutic strategies have been extremely disappointing. Observations of improved outcomes with physiologic doses of corticosteroid replacement therapy and activated protein C (drotrecogin alfa[activated]) have provided new adjuvant therapies for severe sepsis and septic shock in selected patients. This article reviews the components of sepsis management and discusses the available evidence in support of these recommendations. In addition, there is a discussion of some promising new strategies. PMID- 15133469 TI - Hematopoietic stem cells convert into liver cells within days without fusion. AB - Both plasticity and cell fusion have been suggested to have a role in germ-layer switching. To understand the mechanisms underlying cell fate changes, we have examined a highly enriched population of hematopoietic stem cells (HSCs) in vitro or in vivo in response to injury for liver-specific phenotypic and functional changes. Here we show that HSCs become liver cells when cocultured with injured liver separated by a barrier. Chromosomal analyses and tissue-specific gene and/or protein expression show that microenvironmental cues rather than fusion are responsible for conversion in vitro. We transplanted HSCs into liver-injured mice and observed that HSCs convert into viable hepatocytes with increasing injury. Notably, liver function was restored 2-7 d after transplantation. We conclude that HSCs contribute to the regeneration of injured liver by converting into functional hepatocytes without fusion. PMID- 15133470 TI - The endogenous ligand Stunted of the GPCR Methuselah extends lifespan in Drosophila. AB - Many extracellular signals are transmitted to the interior of the cell by receptors with seven membrane-spanning helices that trigger their effects by means of heterotrimeric guanine-nucleotide-binding regulatory proteins (G proteins). These G-protein-coupled receptors (GPCRs) control various physiological functions in evolution from pheromone-induced mating in yeast to cognition in humans. The potential role of the G-protein signalling system in the control of animal ageing has been highlighted by the genetic revelation that mutation of a GPCR encoded by methuselah extends the lifespan of adult Drosophila flies. How methuselah functions in controlling ageing is not clear. A first essential step towards the understanding of methuselah function is to determine the ligands of Methuselah. Here we report the identification and characterization of two endogenous peptide ligands of Methuselah, designated Stunted A and B. Flies with mutations in the gene encoding these ligands show an increase in lifespan and resistance to oxidative stress. We conclude that the Stunted Methuselah system is involved in the control of animal ageing. PMID- 15133472 TI - Analysis of DNA copy number aberrations in hepatitis C virus-associated hepatocellular carcinomas by conventional CGH and array CGH. AB - To clarify the genetic aberrations involved in the development and progression of hepatitis C virus-associated hepatocellular carcinoma (HCV-HCC), we investigated DNA copy number aberrations (DCNAs) in 19 surgically resected HCCs by conventional CGH and array CGH. Conventional CGH revealed that increases of DNA copy number were frequent at 1q (79% of the cases), 8q (37%), 6p (32%), and 10p (32%) and that decreases were frequent at 17p (79%), 16q (58%), 4q (53%), 13q (42%), 10q (37%), 1p (32%), and 8p (32%). In general, genes that showed DCNAs by array CGH were usually located in chromosomal regions with DCNAs detected by conventional CGH analysis. Increases in copy numbers of the LAMC2, TGFB2, and AKT3 genes (located on 1q) and decreases in copy numbers of FGR/SRC2 and CYLD (located on 1p and 16q, respectively) were observed in more than 30% of tumors, including small, well-differentiated carcinomas. These findings suggest that these genes are associated with the development of HCV-HCC. Increases of MOS, MYC, EXT1, and PTK2 (located on 8q) were detected exclusively in moderately and poorly differentiated tumors, suggesting that these alterations contribute to tumor progression. In conclusion, chromosomal and array CGH technologies allow identification of genes involved in the development and progression of HCV-HCC. PMID- 15133471 TI - Imaging the pharmacodynamics of HER2 degradation in response to Hsp90 inhibitors. AB - The development of therapeutic inhibitors of key signaling pathways has been hampered by the inability to assess the effect of a drug on its target in the patient. 17-allylaminogeldanamycin (17-AAG) is the first Hsp90 inhibitor to be tested in a clinical trial. It causes the degradation of HER2 and other Hsp90 targets, and has antitumor activity in preclinical models. We have developed a method for imaging the inhibition of Hsp90 by 17-AAG. We labeled an F(ab')2 fragment of the anti-HER2 antibody Herceptin with 68Ga, a positron emitter, which allows the sequential positron-emission tomographic imaging of HER2 expression. We have used this method to quantify as a function of time the loss and recovery of HER2 induced by 17-AAG in animal tumors. This approach allows noninvasive imaging of the pharmacodynamics of a targeted drug and will facilitate the rational design of combination therapy based on target inhibition. PMID- 15133473 TI - Immunohistochemical detection of ZAP-70 in 341 cases of non-Hodgkin and Hodgkin lymphoma. AB - Using immunohistochemical methods, we evaluated zeta-associated protein (ZAP)-70 expression in 341 cases of non-Hodgkin and Hodgkin lymphoma. In B-cell NHL, ZAP 70 was positive in five of six (83%) precursor B-lymphoblastic lymphoma, 11 of 37 (30%) chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL), five of 39 (13%) mantle cell lymphoma, one of 12 (8%) Burkitt lymphoma, and one of 12 (8%) nodal marginal zone B-cell lymphoma. In 22 cases of CLL/SLL, seven of nine (78%) with unmutated IgVH genes expressed ZAP-70, compared with one of 13 (8%) with mutated IgVH genes (P=0.0015 Fisher's exact test). ZAP-70 expression was not detected in diffuse large B-cell lymphoma (n=26), extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (n=24), follicular lymphoma (n=21), plasma cell myeloma/plasmacytoma (n=10), lymphoplasmacytic lymphoma (n=10), or splenic marginal zone lymphoma (n=6). In T/NK-cell NHL, ZAP-70 was positive in all extranodal natural killer (NK) / T-cell lymphoma, nasal-type (n=6) and enteropathy-type T-cell lymphoma (n=4), four of five (80%) subcutaneous panniculitis-like T-cell lymphoma, six of eight (75%) mycosis fungoides, three of five (60%) precursor T-lymphoblastic lymphoma, 10 of 17 (59%) peripheral T-cell lymphoma, two of four (50%) blastic NK-cell lymphoma, one of three (33%) T-cell prolymphocytic leukemia, 13 of 52 (25%) anaplastic large cell lymphoma, and one of six (17%) angioimmunoblastic T-cell lymphoma. Seven of 12 (58%) cutaneous CD30 positive lymphoproliferative disorders were also ZAP-70-positive. In Hodgkin lymphoma, ZAP-70 was negative in neoplastic cells in all cases tested. ZAP-70 staining in B-cell lymphomas and reactive T cells was predominantly nuclear with variable cytoplasmic staining. By contrast, ZAP-70 staining in T/NK-cell lymphomas was heterogeneous, and a shift from predominantly nuclear to predominantly cytoplasmic staining was observed, particularly in those neoplasms with high-grade morphology. In summary, ZAP-70 is expressed by many lymphoma types, correlates with immunoglobulin heavy-chain variable region gene mutational status in CLL/SLL, and can be detected reliably using immunohistochemical methods. PMID- 15133474 TI - Allelic loss of tumor suppressor genes in ameloblastic tumors. AB - Ameloblastoma is an odontogenic tumor with a variety of histologic appearances and an unpredictable biologic behavior. Little is known about allelic losses of tumor suppressor genes in ameloblastomas. This study surveyed DNA damage in ameloblastomas and correlated this with histologic sub-type and clinical outcome. There were 12 ameloblastomas (two peripheral, eight solid, and two unicystic) and three ameloblastic carcinoma studied for loss of heterozygosity of tumor suppressor genes on chromosomes 1p, 3p, 9p,10q, and 17p (L-myc, hOGG1, p16, pten, and p53). The frequency of allelic loss and the intratumoral heterogeneity were calculated. L-myc (71% frequency of allelic loss) and pten (62% frequency of allelic loss) had the most frequent allelic losses. Overall frequency of allelic loss and intratumoral heterogeneity were higher in mandibular and in unicystic tumors and lower in tumors that recurred/metastasized. The rate of allelic loss in the three carcinomas was similar to that seen in benign tumors. The frequency of allelic loss and intratumoral heterogeneity did not correlate with age, gender, histologic subtype, or prognosis. Since tumors that behaved aggressively did not harbor more allelic losses, it is likely that DNA damage in ameloblastomas and ameloblastic carcinomas is sporadic and cumulative. We conclude that other genetic or epigenetic mechanisms may be responsible for malignant behavior in ameloblastic carcinomas. PMID- 15133475 TI - Nonspecific interstitial pneumonia and usual interstitial pneumonia with mutation in surfactant protein C in familial pulmonary fibrosis. AB - Nonspecific interstitial pneumonia, a recently described form of idiopathic interstitial pneumonia, is characterized by uniform involvement of the alveolar septae with interstitial inflammation and variable amounts of fibrosis. Histological observations differentiate nonspecific interstitial pneumonia from usual interstitial pneumonia and clinically, patients with a nonspecific interstitial pneumonia pattern show better prognosis than those with usual interstitial pneumonia. We have genetically analyzed a family with a history of usual interstitial pneumonia. Most of the patients presented as adults and their biopsies showed a pattern consistent with usual interstitial pneumonia. However, three family members presented in early childhood and their biopsies revealed a nonspecific interstitial pneumonia pattern. The inheritance pattern of usual interstitial pneumonia is consistent with autosomal dominant inheritance with variable expression. DNA sequence analyses of the surfactant protein C gene in children with nonspecific interstitial pneumonia and adults with usual interstitial pneumonia exhibit a common heterozygous mutation located in exon 5. The mutation causes a Leu188 to Gln188 change in the carboxy-terminal region of prosurfactant protein C, possibly affecting peptide processing. These observations suggest that individuals with this particular mutation in surfactant protein C gene might be at increased risk of interstitial lung disease of variety of types. PMID- 15133476 TI - Expression of S100A4 combined with reduced E-cadherin expression predicts patient outcome in malignant melanoma. AB - The aim of the present study was to analyze the expression of S100A4 and E cadherin in a panel of primary and metastatic malignant melanoma, and to correlate the expression level to clinicopathological parameters. The expression of S100A4 was examined by immunohistochemistry in 99 superficial spreading and 60 nodular primary melanomas, while the expression of E-cadherin was analyzed in 92 superficial spreading and 52 nodular lesions from the same panel. The expression levels of S100A4 and E-cadherin in the biopsies were inversely correlated, with S100A4 being expressed at the highest frequency in the nodular and E-cadherin in the superficial spreading lesions, respectively. When analyzing the melanoma subgroups separately, it was revealed that expression of S100A4 had a more significant impact on patient outcome in early superficial spreading melanomas than in the nodular subtype, while E-cadherin expression did not predict patient outcome in any of the subgroups. When examining all the patients, both markers give clinical information as predictors for disease-free survival, but when combining the expression of the two markers, a stronger significant correlation between high E-cadherin expressing/S100A4 negative biopsies and increased disease free survival (P=0.002) was revealed, demonstrating the importance of examining the expression of more than one factor involved in the metastatic cascade when predicting patient outcome. We have also evaluated the relationship between the expression of these two antigens and cell cycle and signal transduction factors. PMID- 15133477 TI - Bronchial margins in lung cancer resection specimens: utility of frozen section and gross evaluation. AB - Pathology reports for all lobectomy and pneumonectomy specimens at UNC Hospitals between 1991 and 2000 (n=405) were reviewed for correlation between frozen section and final bronchial margin, gross distance between tumor and margin and tumor type. Frozen section was performed in 268 cases (66%). A total of 243 were true negatives (90.6 %), 16 (6.0%) were true positives, four (1.5%) were false positives and five (1.9%) were false negatives. The site of tumor in true positive cases was mucosal (11), submucosal (three), lymphatics (one), peribronchial (one). The site of tumor in false-negative cases was submucosal (two), lymphatics (one), peribronchial (two). In 137 cases, no bronchial frozen section was performed; there was one case (0.7%) with positive margin. There was no correlation between final margin positivity and distance between gross tumor and margin. Tumor distance to margin in positive margin cases varied from grossly involved to 3 cm away. There were 72 cases in which wedge resection was performed before lobectomy in which no gross tumor remained in the lobectomy, and in all cases final bronchial margins were negative. In all, 373 of cases (92%) were nonsmall carcinomas. Of these, 10 (2.7%) had positive margins. Tumors other than nonsmall cell carcinoma accounted for a disproportionate number of positive margins. In all, 3/6 of adenoid cystic/mucoepidermoid carcinoma, 1/7 small cell carcinoma and 1/1 lymphoma cases had positive margins. In conclusion, frozen section evaluation of bronchial margins is helpful in central lung tumors. Mucosal tumor is preferentially identified in frozen section. Gross evaluation of margins is problematic, as intramucosal carcinoma or tumor in lymphatics may not be detected, but 3 cm was a 'safe' distance for gross tumor from margin. In lobectomies following wedge resection in which no gross tumor remained, all had negative margins. Salivary gland-type tumors have a high incidence of positive margins, and frozen section is particularly indicated in these tumors. PMID- 15133478 TI - Expression of cytokeratin by malignant meningiomas: diagnostic pitfall of cytokeratin to separate malignant meningiomas from metastatic carcinoma. AB - Based on clinical and histologic features, differentiating metastatic carcinomas from benign or malignant meningiomas usually is not difficult. Occasionally, however, in some patients without a clinical history of carcinoma, malignant meningiomas can morphologically simulate metastatic carcinoma, necessitating an immunohistochemical study for cytokeratin to make a correct diagnosis. However, the utility of immunohistochemical markers to separate malignant meningioma from metastatic carcinoma has not been investigated. The immunoperoxidase method with antigen retrieval was used to characterize the expression of three cytokeratins (AE1/AE3, CAM 5.2, and Pan cytokeratin), EMA, CEA, Ber-EP4, CD 15, and B72.3 in 12 previously diagnosed malignant meningiomas, 20 benign meningiomas, and 20 metastatic carcinomas. Cytokeratin expression was detected in 75% of malignant meningiomas, 0% of benign meningiomas, and 100% of metastatic carcinomas. While epithelial markers of Ber-EP4, CEA, B72.3 and CD-15 were positive in 90, 80, 70 and 65% of the metastatic carcinoma, respectively, they were negative in all 12 malignant meningioma examined. Vimentin immunoreactivity was seen in all benign and malignant meningiomas, and in 20% of metastatic carcinomas. Our results indicated that cytokeratin is not a reliable immunohistochemical marker to separate a malignant meningioma from metastatic carcinoma. A panel of epithelial markers including Ber-EP4, CEA, B72.3 and CD-15, and vimentin may be needed to separate malignant meningioma from metastatic carcinoma. Cytokeratin expression can be a potential pitfall for confusing a malignant meningioma with a metastatic carcinoma. PMID- 15133479 TI - Alterations of DNA mismatch repair proteins and microsatellite instability levels in gastric cancer cell lines. AB - Alterations in DNA mismatch repair (MMR) proteins result in microsatellite instability (MSI), increased mutation accumulation at target genes and cancer development. About one-third of gastric cancers display high-level microsatellite instability (MSI-High) and low-level microsatellite instability (MSI-Low) is frequently detected. To determine whether variations in the levels of MMR proteins or mutations in the main DNA MMR genes are associated with MSI-Low and MSI-High in gastric cancer cell lines, the MSI status (MSI-High, MSI-Low or MS Stable (MSS)) of 14 gastric cancer lines was determined using multiple clone analysis with a panel of five microsatellite markers. Protein levels of hMLH1, hMSH2, hMSH6, hPMS2 and hPMS1 were determined by Western blot. Sequence analysis of hMLH1 and hMSH2 was performed and the methylation status of the hMLH1 promoter was examined. The cell lines SNU1 and SNU638 showed MSI-High, decreased to essentially absent hMLH1 and hPMS2 and reduced hPMS1 and hMSH6 protein levels. The hMLH1 promoter region was hypermethylated in SNU638 cells. The MKN28, MKN87, KATOIII and SNU601 cell lines showed MSI-Low. The MMR protein levels of cells with MSI-Low status was similar to the levels detected in MSS cells. A marked decrease in the expression levels of MutL MMR proteins (hMLH1, hPMS2 and hPMS1) is associated with high levels of MSI mutations in gastric cancer cells. Gastric cancer cell lines with MSI-Low status do not show significant changes in the levels of the main DNA MMR proteins or mutations in the DNA mismatch repair genes hMSH2 and hMLH1. These well-characterized gastric cancer cell lines are a valuable resource to further our understanding of DNA MMR deficiency in cancer development, progression and prognosis. PMID- 15133480 TI - Separable stripe enhancer elements for the pair-rule gene hairy in the beetle Tribolium. AB - Pair-rule genes in Drosophila integrate the positional information provided by the transcription factor gradients of gap genes and set the framework for the expression of the segment polarity genes. Pair-rule genes are also expressed during the segmentation of the short germ-band embryo of the flour beetle Tribolium, although this occurs under cellular conditions that should not allow the generation of transcription factor gradients by passive diffusion. To analyse the regulation of the pair-rule gene hairy in Tribolium, we have used germline transformation with reporter gene constructs based on the piggyBac vector. We can identify an upstream fragment that drives the full expression of the eight pair rule stripes, which are sequentially generated during embryogenesis. Further experiments with smaller fragments reveal separable regions driving stripes three to five, and one region driving stripe three only. Our results suggest that the generation of pair-rule stripes in Tribolium is comparable to that in Drosophila despite the cellular versus syncytial mode of embryogenesis. PMID- 15133481 TI - Combinatorial SNARE complexes with VAMP7 or VAMP8 define different late endocytic fusion events. AB - Both heterotypic and homotypic fusion events are required to deliver endocytosed macromolecules to lysosomes and remodel late endocytic organelles. A trans-SNARE complex consisting of Q-SNAREs syntaxin 7, Vti1b and syntaxin 8 and the R-SNARE VAMP8 has been shown by others to be responsible for homotypic fusion of late endosomes. Using antibody inhibition experiments in rat liver cell-free systems, we confirmed this result, but found that the same Q-SNAREs can combine with an alternative R-SNARE, namely VAMP7, for heterotypic fusion between late endosomes and lysosomes. Co-immunoprecipitation demonstrated separate syntaxin 7 complexes with either VAMP7 or VAMP8 in solubilized rat liver membranes. Additionally, overexpression of the N-terminal domain of VAMP7, in cultured fibroblastic cells, inhibited the mixing of a preloaded lysosomal content marker with a marker delivered to late endosomes. These data show that combinatorial interactions of SNAREs determine whether late endosomes undergo homotypic or heterotypic fusion events. PMID- 15133483 TI - Risk factors for late cytomegalovirus infection after allogeneic stem cell transplantation using HLA-matched sibling donor: donor lymphocyte infusion and previous history of early CMV infection. AB - An increased incidence of late cytomegalovirus (CMV) infection has been reported during the last decade since the introduction of ganciclovir (GCV) prophylaxis or GCV pre-emptive therapy. Given that a donor lymphocyte infusion (DLI) can induce more severe GVHD, this may predispose a patient to late CMV infection. In all, 64 patients (median age 36, M/F 38/26) underwent allogeneic stem cell transplantation (SCT) using a matched sibling donor with bone marrow (n=9) or peripheral blood stem cells (n=55). The overall incidence of CMV infection, early and late CMV infection was 46.9 (30/64), 42.2 (27/64), and 16.4% (9/55), respectively. Early CMV infection was treated with GCV pre-emptive therapy that produced a 92.6% success rate. Among the 20 patients who received 35 DLIs, late CMV infection developed in eight (42.1%) of 19 evaluable cases with a median onset at 127 days post transplant. Risk factors for late CMV infection in a logistic regression analysis included DLIs (P=0.001) and a previous history of CMV infection (P=0.006). In conclusion, late CMV infection was strongly associated with DLIs and a previous history of early CMV infection. Accordingly, extended surveillance of CMV antigenemia is recommended for patients receiving DLIs or who have a previous history of CMV infection. PMID- 15133482 TI - Sgt1 is required for human kinetochore assembly. AB - Budding yeast Sgt1 is required for kinetochore assembly, and its homologues have a role in cAMP signalling in fungi and pathogen resistance in plants. The function of mammalian Sgt1 is unknown. We report that RNA interference-mediated depletion of Sgt1 from HeLa cells causes dramatic alterations of the mitotic spindle and problems in chromosome alignment. Cells lacking Sgt1 undergo a mitotic delay due to activation of the spindle checkpoint. The checkpoint response, however, is significantly weakened in Sgt1-depleted cells, and this correlates with a dramatic reduction in kinetochore levels of Mad1, Mad2 and BubR1. These effects are explained by a problem in kinetochore assembly that prevents the localization of Hec1, CENP-E, CENP-F, CENP-I, but not CENP-C, to mitotic kinetochores. Our studies implicate Sgt1 as an essential protein and a critical assembly factor for the mammalian kinetochore, and lend credit to the hypothesis of a kinetochore assembly pathway that is conserved from yeast to man. PMID- 15133484 TI - High-dose cyclophosphamide with or without etoposide for mobilization of peripheral blood progenitor cells in patients with multiple myeloma: efficacy and toxicity. AB - The purpose of the study was to examine the yield of CD34(+) cells, response rates, and toxicity of high-dose cyclophosphamide with or without etoposide in patients with multiple myeloma. In total, 77 myeloma patients received either cyclophosphamide 4.5 g/m(2) (n=28) alone or with etoposide 2 g/m(2) (n=49) in a nonrandomized manner, followed by G-CSF 10 microg/kg/day for the purpose of stem cell mobilization. The effects of various factors on CD34(+) cell yield, response rate and engraftment were explored. A median of 22.39 x 10(6) CD34(+) cells/kg were collected on the first day of leukapheresis (range 0.59-114.71 x 10(6)/kg) in 71 (92%) of patients. Greater marrow plasma cell infiltration (P=0.02) or prior radiation therapy (P=0.02) adversely affected CD34(+) cell yield. In total, 45% of patients receiving cyclophosphamide and 56% of those receiving cyclophosphamide/etoposide had at least a minimum response by EBMT criteria. In all, 25% of patients who received cyclophosphamide alone vs 75.5% of patients who received combined chemotherapy required hospitalization mainly for treatment of neutropenic fever. Cyclophosphamide alone is associated with impressive CD34(+) cell yields and clear antimyeloma activity. The addition of etoposide resulted in increased toxicity without significant improvement in CD34(+) cell yield or response rates. PMID- 15133485 TI - Graft-versus-myeloma effect following antithymocyte globulin-based reduced intensity conditioning allogeneic stem cell transplantation. AB - In all, 41 multiple myeloma (MM) patients received an antithymocyte globulin (ATG), fludarabine, and busulfan-based reduced intensity conditioning (RIC) for allogeneic stem cell transplantation (allo-SCT) from HLA-identical siblings. In total, 29 patients (70%) were in partial remission, one patient in complete remission, and 11 (27%) with progressive disease at the time of allo-SCT. Median time between diagnosis and allo-SCT was 24 months. The cumulative incidences of grade II-IV and grade III-IV acute graft-versus-host disease (GVHD) were 36% (95% CI, 21-51%) and 7% (95% CI, 2-20%), respectively. Overall, 10 patients developed limited chronic GVHD, whereas seven developed an extensive form (cumulative incidence, 41% (95% CI, 26-56%) at 2 years). With a median follow-up of 389 days, the overall cumulative incidence of transplant-related mortality (TRM) was 17% (95% CI, 6-28%). In all, 11 patients (27%) are in continuous complete remission, and the Kaplan-Meier estimates of overall survival (OS) and progression-free survival (PFS) at 2 years were 62% (95% CI, 47-76%) and 41% (95% CI, 23-62%), respectively. PFS and OS were significantly higher in patients with chronic GVHD as compared to patients without chronic GVHD (P=0.006 for PFS and P=0.01 for OS). Collectively, these data demonstrate that RIC allo-SCT can mediate a potentially curative graft-versus-myeloma effect with an acceptable incidence of toxicity and TRM. PMID- 15133486 TI - Transplantation outcome in allogeneic PBSCT patients according to a new chronic GVHD grading system, including extensive skin involvement, thrombocytopenia, and progressive-type onset. AB - Patients who had suffered chronic GVHD after an allogeneic PBSCT were evaluated using a new chronic GVHD grading system. The study included 36 consecutive adult patients with hematological diseases, who survived at least until day 90 following allogeneic PBSCT and who could be evaluated for chronic GVHD. Extensive skin involvement was observed in five patients, thrombocytopenia in 14, and progressive-type onset in 10, while grade 1 chronic GVHD appeared in 21 patients, grade 2 in 10, and grade 3 in five. There was a significant difference in the probability of relapse between the groups with grade 1 and 2+3 chronic GVHD (55.3 vs 16.4%, P=0.0211). The difference was particularly marked in patients with high risk hematological malignancies (grade 1 vs grade 2+3, 75 vs 0%, P=0.0115). With a median follow-up of 12 months (range, 4-52 months), 22 (66.1%) patients were still alive. The estimated 2-year survival rate for the whole population was 57.6%, while that for the group with chronic GVHD grade 1 and grade 2+3 was 53.5 and 56.3%, respectively (P=0.4387). Accordingly, there was a significant difference in the probability of relapse between the groups with grade 1 and grade 2+3 chronic GVHD. PMID- 15133487 TI - CD8+ cell depletion of donor lymphocyte infusions using cd8 monoclonal antibody coated high-density microparticles (CD8-HDM) after allogeneic hematopoietic stem cell transplantation: a pilot study. AB - A CD8 murine monoclonal antibody-coated high-density microparticle (HDM) has been developed, which allows for the rapid depletion of CD8+ T cells from apheresis products by gravity sedimentation. We conducted a study to determine the efficacy and safety of CD8 depletion of donor lymphocyte infusions (DLI) to treat relapse after stem cell transplantation using the Eligix CD8-HDM Cell Separation System. Patients were targeted to receive 3 x 10(7) CD4+ T cells/kg. Nine patients were enrolled, three with CML, three myeloma, two CLL, and one NHL. A median of 1 x 10(10) mononuclear cells were obtained by apheresis and processed. The median depletion of CD8+ cells was 99.3% (97.8->99.5%). CD8 depletion was highly specific, with a median recovery of CD4+ cells of 75%. A median of 2.9 x 10(7) CD4+ cells/kg was infused. No infusional toxicity was noted. All CML patients achieved a complete molecular remission. A CLL patient demonstrated a complete response. One patient developed GVHD (grade II acute GVHD and subsequently chronic GVHD). The CD8-HDM Cell Separation System appears to be highly selective and effective in depleting CD8+ T cells from DLI apheresis products, and CD8 depleted DLI is capable of mediating a graft-versus-leukemia effect while minimizing GVHD. PMID- 15133488 TI - Conjunctival carcinoma as a novel post-stem cell transplantation malignancy. PMID- 15133489 TI - High-dose therapy and autologous stem cell transplantation for multiple myeloma poorly responsive to initial therapy. AB - Autologous stem cell transplant (SCT) improves survival in multiple myeloma (MM) and remains the standard of care for eligible patients. Nearly a third of patients with newly diagnosed MM fail initial therapy aimed at reducing tumor burden preceding SCT (primary refractory). It is unclear if an initial response is important for successful SCT. We evaluated our experience with SCT in 50 patients with primary refractory MM and compared it to 101 patients with chemosensitive disease receiving SCT. The study cohort had a median age of 56 years (range 29-72) consisting of 87 males (58%). A total of 46 patients (92%) in the refractory group and 100 (99%) in the chemosensitive group had a response to transplant (50% or greater reduction in the M-protein). In all, 10 refractory patients (20%) and 35 (35%) in the chemosensitive group achieved a CR (P=0.06). The 1-year estimated progression-free survival from the time of transplant for the refractory group was 70% compared to 83% for the chemosensitive group (P=0.65). The lack of response to initial induction therapy does not appear to preclude a good response to SCT. We recommend that patients with primary refractory MM be offered early SCT. PMID- 15133490 TI - CCND1 polymorphism and age of onset of hepatoblastoma. AB - Cyclin D1, encoded by the gene CCND1, is a major regulator of the cell cycle transition from G1 phase to S phase. A CCND1 polymorphism (G to A) at codon 242, the boundary of exon 4 and intron 4, affects splicing such that exon 5 is not expressed in the A allele. Since exon 5 is involved in rapid turnover, the variant cyclin D1 corresponding to the A allele may have a longer half-life. A previous study demonstrated that in families with hereditary nonpolyposis colorectal cancer, the age of onset of colorectal cancer varied according to variation at this polymorphic site. We examined this CCND1polymorphism in a series hepatoblastoma, a childhood liver cancer that shares other molecular features with colon cancer. We determined in an analysis of 84 children with hepatoblastoma that the G/A exon 4 polymorphism in CCND1 is correlated with the age of onset of hepatoblastomas. The A/A genotype is associated with an earlier age of onset compared to the G/A or G/G genotype. The median age of patients with the G/G genotype was 22 months, compared to 17 months in patients with the G/A genotype and 11 months for the A/A genotype. These findings suggest that the CCND1 A polymorphism may contribute to tumor development in children with hepatoblastoma. PMID- 15133491 TI - Genetic and epigenetic alterations of the APC gene in malignant melanoma. AB - High levels of beta-catenin and activating mutations in the beta-catenin gene (CTNNB1) have been demonstrated in malignant melanomas, implicating dysregulated Wnt signalling in the pathogenesis of this malignancy. We systematically examined melanoma cell lines for activating CTNNB1 mutations as well as genetic and epigenetic alterations of the adenomatous polyposis coli gene (APC), another key component of the Wnt signalling transduction pathway. Of 40 cell lines tested, one carried a truncating APC mutation and loss of the corresponding wild-type allele, and one carried a CTNNB1 missense mutation. Hypermethylation of APC promoter 1A was present in five of the cell lines (13%) and in nine of 54 melanoma biopsies (17%). Cells with truncating APC or activating CTNNB1 mutations showed increased transcription from endogenous and ectopic beta-catenin/T-cell factor (Tcf)-responsive target genes, consistent with the known effects of these alterations on beta-catenin stability and Tcf transactivation. In contrast, cell lines with APC promoter 1A hypermethylation did not show increased Wnt signalling, probably due to residual APC activity expressed from promoter 1B. Suppression of APC transcripts in melanoma cells by stable expression of short hairpin RNAs led to a Wnt signalling-independent increase in cell proliferation, but also reduced the invasive growth in collagen type I. Collectively, our data suggest that the tumour-suppressive function of APC in melanocytic cells is dose dependent. We propose that epigenetic silencing of promoter 1A may contribute to the development of malignant melanoma by reducing the expression of APC to a level that promotes cell proliferation without compromising the invasive capacity. PMID- 15133493 TI - Transforming growth factor beta regulates cell-cell adhesion through extracellular matrix remodeling and activation of focal adhesion kinase in human colon carcinoma Moser cells. AB - Transforming growth factor (TGF) beta is a potent regulator of cell-matrix and cell-cell adhesions (collectively termed cellular adhesions). Cellular adhesions play crucial roles in controlling the differentiation of epithelial cells and in maintaining the integrity of the epithelium. Loss of TGF beta-responsiveness is thought to be an important early initiating event in the malignant progression of epithelial cancer. In the TGFbeta-responsive human colon adenocarcinoma Moser cells, TGFbeta promotes cellular adhesions and suppresses their malignant phenotype. TGFbeta promotes cell-matrix adhesion by inducing the synthesis of extracellular matrix (ECM) adhesion molecules and the expression of integrin receptors for these molecules (termed ECM remodeling). TGFbeta promotes cell-cell adhesion through the induction of E-cadherin expression, an epithelial associated homotypic cell-cell adhesion molecule, which also functions as a tumor suppressor in colon cancer. How TGFbeta regulates E-cadherin expression is not known. In this study, we showed that the induction of E-cadherin by TGFbeta was mediated through the activation of focal adhesion kinase (FAK), a major signaling molecule in focal adhesion contacts and that the activation of FAK was due to ECM remodeling and increased cell-matrix interactions. Thus, TGFbeta regulates cell cell adhesion through its ability to remodel the ECM and to activate FAK through ECM remodeling. PMID- 15133494 TI - JCV T-antigen interacts with the neurofibromatosis type 2 gene product in a transgenic mouse model of malignant peripheral nerve sheath tumors. AB - The human polyomavirus, JC virus, has recently been associated with several human CNS tumors, including medulloblastomas and a broad range of glial-origin tumors. This ubiquitous virus is the causative agent of the rare demyelinating disease, progressive multifocal leukoencephalopathy in immunocompromised individuals. Expression of the viral protein, T-antigen, which possesses the ability to transform cells of neural origin, has been detected in human CNS tumors. In an effort to further understand the transforming potential of JCV T-antigen, transgenic mice expressing JCV T-antigen under the control of the Mad-4 promoter were generated. As described previously, approximately 50% of the animals developed pituitary tumors by 1 year of age. However, a small subset of the animals developed solid masses arising from the soft tissues surrounding the salivary gland, the sciatic nerve, and along the extremities that histologically resemble malignant peripheral nerve sheath tumors, rare neoplasms that occur in individuals with neurofibromatosis type 1 (NF1). JCV T-antigen was detected in tumor tissue by immunohistochemistry and immunoprecipitation/Western blotting, but not in normal tissues and was colocalized with NF2, the putative tumor suppressor protein associated with neurofibromatosis type 2, in the nucleus of some cells. In addition, T-antigen was co-precipitated with NF2, but not with NF1 protein, although NF1 was detectable in tumor tissue. Furthermore, precipitated immunocomplexes contained T-antigen, NF2, and p53, suggesting that these three proteins may form a ternary complex. The importance of these findings on mechanisms of T-antigen-mediated tumorigenesis and the pathogenesis of neurofibromatosis are discussed. Oncogene (2004) 23, 5459-5467. doi:10.1038/sj.onc.1207728 Published online 10 May 2004 PMID- 15133492 TI - E2F-7: a distinctive E2F family member with an unusual organization of DNA binding domains. AB - The E2F family of transcription factors play an important role in regulating cell cycle progression. We report here the characterization and functional properties of a new member of the human E2F family, referred to as E2F-7. E2F-7 has two separate DNA-binding domains, a feature that distinguishes E2F-7 from other mammalian E2F proteins, but resembling the organization of recently isolated E2F like proteins from Arabidopsis. E2F-7 binds to DNA independently of a DP partner and delays cell cycle progression. Interestingly, E2F-7 modulates the transcription properties of other E2F proteins. A mutational analysis indicates that the integrity of both DNA-binding domains is required for cell cycle delay and transcriptional modulation. Biochemical results and protein modelling studies suggest that in binding to DNA interactions occur between the two DNA-binding domains, most probably as a homodimer, thereby mimicking the organization of an E2F/DP heterodimer. These structural and functional properties of E2F-7 imply a unique role in regulating cellular proliferation. PMID- 15133496 TI - FAPalpha, a surface peptidase expressed during wound healing, is a tumor suppressor. AB - Fibroblast activation protein-alpha (FAP) is a cell surface serine protease expressed at sites of tissue remodeling in embryonic development. FAP is not expressed by mature somatic tissues except activated melanocytes and fibroblasts in wound healing or tumor stroma. FAP expression is specifically silenced in proliferating melanocytic cells during malignant transformation. To study the role of FAP as a tumor suppressor, the gene for mouse fap was cloned and mutated at the catalytic domain (FAP serine mutant, FSM). We found that expression of FAP or FSM at physiologic levels in mouse melanoma cells abrogated tumorigenicity. Remarkably, the mutant form FSM lacking specific serine protease activity was a more potent tumor suppressor. Tumor rejection was not due to adaptive immune responses because RAG1-/- mice challenged with melanoma cells expressing either FAP or FSM were not tumorigenic. In in vitro assays, FAP or FSM expression restored contact inhibition, led to cell cycle arrest at G0/G1 phase, and increased susceptibility to stress-induced apoptosis. Cell death in FAP+ or FSM+ melanoma cells was readily triggered by depletion of survival factors from the media, leading to subsequent activation of caspases via the intrinsic pathway. These results show that expression of FAP is a tumor suppressor that abrogates tumorigenicity through regulation of cell growth and survival. PMID- 15133495 TI - RUNX1 transformation of primary embryonic fibroblasts is revealed in the absence of p53. AB - The mammalian Runx gene family (Runx1-3) are transcription factors that play essential, lineage-specific roles in development. A growing body of evidence implicates these genes as mutational targets in cancer where, in different contexts, individual family members have been reported to act as tumour suppressors, dominant oncogenes or mediators of metastasis. We are exploring these paradoxical observations by ectopic expression of RUNX genes in primary murine embryonic fibroblasts where, in common with a number of other dominant oncogenes, RUNX1 induces senescence-like growth arrest in the presence of an intact p19(ARF)-p53 pathway. We now report that, in MEFs lacking functional p53, RUNX1 has apparently pro-oncogenic effects on cell growth that include cytoskeletal reorganization, reduced contact inhibition at confluence and accelerated tumour expansion in vivo. On the other hand, RUNX1 conferred no obvious growth advantage at low cell density and actually delayed entry of primary MEFs into S phase. We also found that ectopic RUNX1 interferes with the morphological and growth responses of p53-null MEFs to TGFbeta indicating that these effects are mediated by overlapping pathways. These observations help to elucidate the context-dependent consequences of loss and gain of Runx activity. PMID- 15133497 TI - v-Jun targets showing an expression pattern that correlates with the transformed cellular phenotype. AB - Targets of the oncogenic transcription factor v-Jun in the murine cell line C3H 10T1/2 cells have been identified using DNA microarrays. Two targets, Akap12 and Marcks, are downregulated in transformed cells and are known tumor suppressor genes. Overexpression of either Akap12 or Marcks in v-Jun-transformed cells reverses the transformed phenotype and leads to the re-expression of the other tumor suppressor gene, suggesting that these two genes cooperate in the establishment of the nontransformed state. Reverted cells continue to express v Jun at high levels and also re-express c-Jun, which is normally repressed by v Jun. A panel of six cell lines has been generated to evaluate the expression levels of other v-Jun targets in 10T1/2 cells. With these cells, we find that the upregulated target Sprr1a has an expression pattern that correlates with the transformed phenotype. PMID- 15133498 TI - An activated mTOR mutant supports growth factor-independent, nutrient-dependent cell survival. AB - In yeast, TOR couples cellular growth and metabolism to the availability of extracellular nutrients. In contrast, mammalian TOR kinase activity has been reported to be regulated by growth factor stimulation via the PI3K/Akt pathway. Consistent with this, growth factor deprivation results in dephosphorylation of the mTOR target proteins p70S6k and 4EBP1 in the face of abundant extracellular nutrients. To determine whether the activation of mTOR was sufficient to support cell survival in the absence of other growth factor-mediated signal transduction, we evaluated the ability of a growth factor-independent mTOR mutant, DeltaTOR, to protect cells from growth factor deprivation. DeltaTOR- but not wild-type mTOR expressing cells were protected from many of the sequelae of growth factor deprivation including amino-acid transporter degradation, reduction of the glycolytic rate, cellular atrophy, decreased mitochondrial membrane potential, and Bax activation. Furthermore, DeltaTOR expression increased growth factor independent, nutrient-dependent cell survival and enhanced the ability of p53-/- MEFs to form colonies in soft agar. These results suggest that activating mutations of mTOR can contribute to apoptotic resistance and might contribute to cellular transformation. PMID- 15133499 TI - Genome-wide analysis of mRNAs regulated by the THO complex in Drosophila melanogaster. AB - In yeast cells, the THO complex has been implicated in mitotic recombination, transcription elongation and mRNA nuclear export. The stable core of THO consists of Tho2p, Hpr1p, Mft1p and Thp2p. Whether a complex with similar functions assembles in metazoa has not yet been established. Here we report that Drosophila melanogaster THO consists of THO2, HPR1 and three proteins, THOC5-THOC7, which have no orthologs in budding yeast. Gene expression profiling in cells depleted of THO components revealed that <20% of the transcriptome was regulated by THO. Nonetheless, export of heat-shock mRNAs under heat stress was strictly dependent on THO function. Notably, 8% of upregulated genes encode proteins involved in DNA repair. Thus, although THO function seems to be conserved, the vast majority of mRNAs are transcribed and exported independently of THO in D. melanogaster. PMID- 15133500 TI - The stalk region of dynamin drives the constriction of dynamin tubes. AB - The GTPase dynamin is essential for numerous vesiculation events including clathrin-mediated endocytosis. Upon GTP hydrolysis, dynamin constricts a lipid bilayer. Previously, a three-dimensional structure of mutant dynamin in the constricted state was determined by helical reconstruction methods. We solved the nonconstricted state by a single-particle approach and show that the stalk region of dynamin undergoes a large conformational change that drives tube constriction. PMID- 15133501 TI - A unique catalytic mechanism for UDP-galactopyranose mutase. AB - The flavoenzyme uridine 5'-diphosphate (UDP)-galactopyranose mutase (UGM) catalyzes the interconversion of UDP-galactopyranose (UDP-Galp) and UDP galactofuranose (UDP-Galf). The latter is an essential precursor to the cell wall arabinogalactan of Mycobacterium tuberculosis. The catalytic mechanism for this enzyme had not been elucidated. Here, we provide evidence for a mechanism in which the flavin cofactor assumes a new role. Specifically, the N5 of the reduced anionic flavin cofactor captures the anomeric position of the galactose residue with release of UDP. Interconversion of the isomers occurs via a flavin-derived iminium ion. To trap this putative intermediate, we treated UGM with radiolabeled UDP-Galp and sodium cyanoborohydride; a radiolabeled flavin-galactose adduct was obtained. Ultraviolet-visible spectroscopy and mass spectrometry indicate that this product is an N5-alkyl flavin. We anticipate that the clarification of the catalytic mechanism for UGM will facilitate the development of anti-mycobacterial agents. PMID- 15133503 TI - Structural basis of phosphopeptide recognition by the BRCT domain of BRCA1. AB - The BRCT repeats in BRCA1 are essential for its tumor suppressor activity and interact with phosphorylated protein targets containing the sequence pSer-X-X Phe, where X indicates any residue. The structure of the tandem BRCA1 BRCT repeats bound to an optimized phosphopeptide reveals that the N-terminal repeat harbors a conserved BRCT phosphoserine-binding pocket, while the interface between the repeats forms a hydrophobic groove that recognizes the phenylalanine. Crystallographic and biochemical data suggest that the structural integrity of both binding sites is essential for peptide recognition. The diminished peptide binding capacity observed for cancer-associated BRCA1-BRCT variants may explain the enhanced cancer risks associated with these mutations. PMID- 15133502 TI - Structure and mechanism of BRCA1 BRCT domain recognition of phosphorylated BACH1 with implications for cancer. AB - Germline mutations in the BRCA1 tumor suppressor gene often result in a significant increase in susceptibility to breast and ovarian cancers. Although the molecular basis of their effects remains largely obscure, many mutations are known to target the highly conserved C-terminal BRCT repeats that function as a phosphoserine/phosphothreonine-binding module. We report the X-ray crystal structure at a resolution of 1.85 A of the BRCA1 tandem BRCT domains in complex with a phosphorylated peptide representing the minimal interacting region of the DEAH-box helicase BACH1. The structure reveals the determinants of this novel class of BRCA1 binding events. We show that a subset of disease-linked mutations act through specific disruption of phospho-dependent BRCA1 interactions rather than through gross structural perturbation of the tandem BRCT domains. PMID- 15133504 TI - Substantial reduction of critical current for magnetization switching in an exchange-biased spin valve. AB - Great interest in current-induced magnetic excitation and switching in a magnetic nanopillar has been caused by the theoretical predictions of these phenomena. The concept of using a spin-polarized current to switch the magnetization orientation of a magnetic layer provides a possible way to realize future 'current-driven' devices: in such devices, direct switching of the magnetic memory bits would be produced by a local current application, instead of by a magnetic field generated by attached wires. Until now, all the reported work on current-induced magnetization switching has been concentrated on a simple ferromagnet/Cu/ferromagnet trilayer. Here we report the observation of current induced magnetization switching in exchange-biased spin valves (ESPVs) at room temperature. The ESPVs clearly show current-induced magnetization switching behaviour under a sweeping direct current with a very high density. We show that insertion of a ruthenium layer between an ESPV nanopillar and the top electrode effectively decreases the critical current density from about 10(8) to 10(7) A cm(-2). In a well-designed 'antisymmetric' ESPV structure, this critical current density can be further reduced to 2 x 10(6) A cm(-2). We believe that the substantial reduction of critical current could make it possible for current induced magnetization switching to be directly applied in spintronic devices, such as magnetic random-access memory. PMID- 15133505 TI - Strain-controlled growth of nanowires within thin-film cracks. AB - There is continued interest in finding quicker and simpler ways to fabricate nanowires, even though research groups have been investigating possibilities for the past decade. There are two reasons for this interest: first, nanowires have unusual properties-for example, they show quantum-mechanical confinement effects, they have a very high surface-to-volume ratio, enabling them to be used as sensors, and they have the ability to connect to individual molecules. Second, no simple method has yet been found to fabricate nanowires over large areas in arbitrary material combinations. Here we describe an approach to the generation of well-defined nanowire network structures on almost any solid material, up to macroscopic sample sizes. We form the nanowires within cracks in a thin film. Such cracks have a number of properties that make them attractive as templates for nanowire formation: they are straight, scalable down to nanometre size, and can be aligned (by using microstructure to give crack alignment via strain). We demonstrate the production of nanowires with diameter <16 nm, both singly and as networks; we have also produced aligned patterns of nanowires, and nanowires with individual contacts. PMID- 15133507 TI - Self-reactivity in thymic double-positive cells commits cells to a CD8 alpha alpha lineage with characteristics of innate immune cells. AB - Thymocytes displaying self-reactive T cell receptors usually undergo negative selection in the thymus. Here we demonstrate that agonist peptides can promote positive selection of immature double-positive thymocytes into distinct lineages, varying with the agonist concentration and the animal's age. Microarray gene expression analyses showed broad transcriptional alterations in a set of transcripts associated with the innate immune system, as well as silencing of CD8 beta expression. The resulting CD8 alpha alpha T cells showed a rapid effector cytokine response. Hence, T cells displaying self-reactive receptors can have the gene expression profile and phenotypic characteristics of innate immune cells. PMID- 15133508 TI - A Bcl-2-dependent molecular timer regulates the lifespan and immunogenicity of dendritic cells. AB - The lifespan of antigen-bearing dendritic cells (DCs) is determined by signals from pathogens and T cells. These signals regulate DC survival by modulating expression of Bcl-2 family proteins. Toll-like receptors and T cell costimulatory molecules both trigger a DC survival pathway that is dependent on Bcl-x(L). However, Toll-like receptors uniquely increase expression of Bim and trigger cell death by a pathway that is blocked by Bcl-2. This pathway serves as a molecular 'timer' that sets the lifespan of DCs and regulates the magnitude of T cell responses in vivo. Thus, signals derived from the innate and acquired immune systems control DC lifespan and immunogenicity by distinct molecular mechanisms. PMID- 15133506 TI - Reversal of obesity by targeted ablation of adipose tissue. AB - Obesity is an increasingly prevalent human condition in developed societies. Despite major progress in the understanding of the molecular mechanisms leading to obesity, no safe and effective treatment has yet been found. Here, we report an antiobesity therapy based on targeted induction of apoptosis in the vasculature of adipose tissue. We used in vivo phage display to isolate a peptide motif (sequence CKGGRAKDC) that homes to white fat vasculature. We show that the CKGGRAKDC peptide associates with prohibitin, a multifunctional membrane protein, and establish prohibitin as a vascular marker of adipose tissue. Targeting a proapoptotic peptide to prohibitin in the adipose vasculature caused ablation of white fat. Resorption of established white adipose tissue and normalization of metabolism resulted in rapid obesity reversal without detectable adverse effects. Because prohibitin is also expressed in blood vessels of human white fat, this work may lead to the development of targeted drugs for treatment of obese patients. PMID- 15133509 TI - CD22 attenuates calcium signaling by potentiating plasma membrane calcium-ATPase activity. AB - Binding of antigen to the B cell receptor induces a calcium response, which is required for proliferation and antibody production. CD22, a B cell surface protein, inhibits this signal through mechanisms that have been obscure. We report here that CD22 augments calcium efflux after B cell receptor crosslinking. Inhibition of plasma membrane calcium-ATPase (PMCA) attenuated these effects, as did disruption by homologous recombination of the gene encoding PMCA4a and PMCA4b. PMCA coimmunoprecipitated with CD22 in an activation-dependent way. CD22 cytoplasmic tyrosine residues were required for association with PMCA and enhancement of calcium efflux. Moreover, CD22 regulation of efflux and the calcium response required the tyrosine phosphatase SHP-1. Thus, SHP-1 and PMCA provide a mechanism by which CD22, a tissue-specific negative regulator, can affect calcium responses. PMID- 15133510 TI - Mutations in SEC63 cause autosomal dominant polycystic liver disease. AB - Mutations in PRKCSH, encoding the beta-subunit of glucosidase II, an N-linked glycan-processing enzyme in the endoplasmic reticulum (ER), cause autosomal dominant polycystic liver disease. We found that mutations in SEC63, encoding a component of the protein translocation machinery in the ER, also cause this disease. These findings are suggestive of a role for cotranslational protein processing pathways in maintaining epithelial luminal structure and implicate noncilial ER proteins in human polycystic disease. PMID- 15133511 TI - Mutations in GALNT3, encoding a protein involved in O-linked glycosylation, cause familial tumoral calcinosis. AB - Familial tumoral calcinosis (FTC; OMIM 211900) is a severe autosomal recessive metabolic disorder that manifests with hyperphosphatemia and massive calcium deposits in the skin and subcutaneous tissues. Using linkage analysis, we mapped the gene underlying FTC to 2q24-q31. This region includes the gene GALNT3, which encodes a glycosyltransferase responsible for initiating mucin-type O glycosylation. Sequence analysis of GALNT3 identified biallelic deleterious mutations in all individuals with FTC, suggesting that defective post translational modification underlies the disease. PMID- 15133512 TI - Recombination and the Tel1 and Mec1 checkpoints differentially effect genome rearrangements driven by telomere dysfunction in yeast. AB - In telomerase-deficient Saccharomyces cerevisiae, telomeres are maintained by recombination. Here we used a S. cerevisiae assay for characterizing gross chromosomal rearrangements (GCRs) to analyze genome instability in post-senescent telomerase-deficient cells. Telomerase-deficient tlc1 and est2 mutants did not have increased GCR rates, but their telomeres could be joined to other DNAs resulting in chromosome fusions. Inactivation of Tel1 or either the Rad51 or Rad59 recombination pathways in telomerase-deficient cells increased the GCR rate, even though telomeres were maintained. The GCRs were translocations and chromosome fusions formed by nonhomologous end joining. We observed chromosome fusions only in mutant strains expressing Rad51 and Rad55 or when Tel1 was inactivated. In contrast, inactivation of Mec1 resulted in more inversion translocations such as the isochromosomes seen in human tumors. These inversion translocations seemed to be formed by recombination after replication of broken chromosomes. PMID- 15133513 TI - TIN2 is a tankyrase 1 PARP modulator in the TRF1 telomere length control complex. AB - Telomere length in humans is partly controlled by a feedback mechanism in which telomere elongation by telomerase is limited by the accumulation of the TRF1 complex at chromosome ends. TRF1 itself can be inhibited by the poly(ADP-ribose) polymerase (PARP) activity of its interacting partner tankyrase 1, which abolishes its DNA binding activity in vitro and removes the TRF1 complex from telomeres in vivo. Here we report that the inhibition of TRF1 by tankyrase is in turn controlled by a second TRF1-interacting factor, TIN2 (ref. 6). Partial knockdown of TIN2 by small hairpin RNA in a telomerase-positive cell line resulted in telomere elongation, which is typical of reduced TRF1 function. Transient inhibition of TIN2 with small interfering RNA led to diminished telomeric TRF1 signals. This effect could be reversed with the PARP inhibitor 3 aminobenzamide and did not occur in cells overexpressing a PARP-dead mutant of tankyrase 1. TIN2 formed a ternary complex with TRF1 and tankyrase 1 and stabilized their interaction, an effect also observed with the PARP-dead mutant of tankyrase 1. In vitro, TIN2 protected TRF1 from poly(ADP-ribosyl)ation by tankyrase 1 without affecting tankyrase 1 automodification. These data identify TIN2 as a PARP modulator in the TRF1 complex and can explain how TIN2 contributes to the regulation of telomere length. PMID- 15133514 TI - Neural mechanism underlying complex receptive field properties of motion sensitive interneurons. AB - In many species, neurons responding to visual motion at higher processing stages are often specifically tuned to particular flow fields; however, the neural circuitry that leads to this selectivity is not yet understood. Here we have studied this problem in 'vertical system' (VS) cells of the blowfly lobula plate. These neurons possess distinctive local preferred directions in different parts of their receptive field. Dual recordings from pairs of VS cells show that they are electrically coupled. This coupling is responsible for the elongated horizontal extent of their receptive fields. VS cells with a lateral receptive field have additional connections to a VS cell with a frontal receptive field and to the horizontal system, tuning these cells to rotational flow fields. In summary, the receptive field of these cells consists of two components: one that they receive from local motion detectors on their dendrite, and one that they import from other large-field neurons. PMID- 15133515 TI - Ascl1/Mash1 is required for the development of central serotonergic neurons. AB - The transcriptional control of the differentiation of central serotonergic (5-HT) neurons in vertebrates has recently come under scrutiny and has been shown to involve the homeobox genes Nkx2-2 and Lmx1b, the Ets-domain gene Pet1 (also known as Fev) and the zinc-finger gene Gata3. The basic helix-loop-helix (bHLH) gene Ascl1 (also known as Mash1) is coexpressed with Nkx2-2 in the neuroepithelial domain of the hindbrain, which gives rise to 5-HT neurons. Here we show in the mouse that Ascl1 is essential for the birth of 5-HT neurons, both as a proneural gene for the production of postmitotic neuronal precursors and as a determinant of the serotonergic phenotype for the parallel activation of Gata3, Lmx1b and Pet1. Thus Ascl1, which is essential for noradrenergic differentiation, is also a determinant of the serotonergic phenotype. PMID- 15133516 TI - Overexpression of type-1 adenylyl cyclase in mouse forebrain enhances recognition memory and LTP. AB - Cyclic AMP is a positive regulator of synaptic plasticity and is required for several forms of hippocampus-dependent memory including recognition memory. The type I adenylyl cyclase, Adcy1 (also known as AC1), is crucial in memory formation because it couples Ca(2+) to cyclic AMP increases in the hippocampus. Because Adcy1 is neurospecific, it is a potential pharmacological target for increasing cAMP specifically in the brain and for improving memory. We have generated transgenic mice that overexpress Adcy1 in the forebrain using the Camk2a (also known as alpha-CaMKII) promoter. These mice showed elevated long term potentiation (LTP), increased memory for object recognition and slower rates of extinction for contextual memory. The increase in recognition memory and lower rates of contextual memory extinction may be due to enhanced extracellular signal related kinase (ERK)/mitogen-activated protein kinase (MAPK) signaling, which is elevated in mice that overexpress Adcy1. PMID- 15133517 TI - The prepattern transcription factor Irx2, a target of the FGF8/MAP kinase cascade, is involved in cerebellum formation. AB - The cerebellum develops from the rhombic lip of the rostral hindbrain and is organized by fibroblast growth factor 8 (FGF8) expressed by the isthmus. Here we report characterization of Irx2, a member of the Iroquois (Iro) and Irx class of homeobox genes, that is expressed in the presumptive cerebellum. When Irx2 is misexpressed with Fgf8a in the chick midbrain, the midbrain develops into cerebellum in conjunction with repression of Otx2 and induction of Gbx2. During this event, signaling by the FGF8 and mitogen-activated protein (MAP) kinase cascade modulates the activity of Irx2 by phosphorylation. Our data identify a link between the isthmic organizer and Irx2, thereby shedding light on the roles of Iro and Irx genes, which are conserved in both vertebrates and invertebrates. PMID- 15133518 TI - A brief history of resuscitation. PMID- 15133519 TI - Multidisciplinary approach to breast disease-now is the time for specialist units in New Zealand. PMID- 15133520 TI - Knowledge and attitudes towards cardiopulmonary resuscitation in the community. AB - AIMS: This study examined basic aspects of knowledge and attitudes towards resuscitation in a New Zealand urban community. METHODS: Using a telephone survey, we questioned 400 subjects aged (over 17 years of age), on their prior training, knowledge, and attitudes towards resuscitation. RESULTS: Seventy-four percent of subjects had previously been taught cardiopulmonary resuscitation (CPR). Of these, 12% had been taught during the previous year, and 63% over 5 years previously. Older subjects were less likely to have learnt CPR than younger subjects. Seventy-three percent of those surveyed desired to know more about resuscitation (than they currently did) and 70% thought that resuscitation should be a compulsory component of the New Zealand Driver's Licence test. Sixty-three percent said they would be willing to perform mouth-to-mouth ventilation on a stranger. CPR knowledge was poor, however, with only 4% knowing an acceptable rate at which to perform chest compressions, and only 9% knowing the correct compression-to-ventilations ratio for adult CPR. Overall knowledge was highest for those taught in the previous year, and for those persons aged between 26 and 45 years. CONCLUSIONS: Although attitudes of the community toward CPR are positive, theoretical knowledge relating to basic CPR is poor. This suggests that present community CPR educational strategies have limited efficacy. PMID- 15133521 TI - Frequent attenders at Christchurch Hospital's Emergency Department: a 4-year study of attendance patterns. AB - AIMS: To describe the pattern of Emergency Department (ED) use by a cohort of adult frequent attenders over a 4-year period. METHODS: A descriptive retrospective cohort study was performed of frequent attenders, identified as having 10 or more presentations to the ED, from 1 January 1997 to 31 December 1997. Diagnoses were assigned from hospital records. ED visits for the subsequent 3 years were recorded. RESULTS: Seventy-seven patients were identified. They made 1127 (1.7%) of the 65,007 ED visits in 1997. Thirty-five patients (45%) had a medical problem, 22 (29%) had a psychiatric problem, and 20 (26%) had a diagnosis of substance abuse as the main reason for their ED visits. Twenty-seven patients (35%) had a diagnosis in more than one category. Of the original 77 patients, 46 (60%) were frequent attenders only in the index year. In the final study year, 13 patients (17%) remained frequent attenders, 37 (48%) presented less than 10 times, and 27 (35%) made zero visits. Seven patients (9%) remained frequent attenders in all 4 years. Ten patients (13%) died. Five (6%) were identified as having left the area. In the final year of the study, there were, by the same criteria, a total of 76 frequent attenders to the ED. Only 13 (17%) of these attenders were from the original cohort. CONCLUSION: Frequent attenders to the ED are not, as commonly assumed, a constant population. The majority of patients change their frequency of attendance over time. Those patients who remain high users have a higher incidence of psychiatric illness and substance abuse. To initiate management intervention, ED records must be regularly monitored to identify the constant stream of new frequent attenders. PMID- 15133522 TI - Respiratory symptoms and lung function change in welders: are they associated with workplace exposures? AB - AIMS: This study investigates whether work-related respiratory symptoms and acute falls in forced expiratory volume in 1 second (FEV1), previously observed in current welders, are related to measured workplace exposures to total fume and metals. METHODS: At four work sites in New Zealand, changes in pulmonary function (and reported respiratory symptoms) were recorded in 49 welding workers (and 26 non-welders) exposed to welding fume. We also determined the personal breathing zone levels of total fume and various metals. RESULTS: Work-related respiratory symptoms were reported by 26.5% of welders and 11.5% of non-welders. These symptoms were related significantly to their personal breathing zone nickel exposure--with an adjusted odds ratio (OR) and 95% confidence interval [CI] of the high exposure group (compared to a low exposure group of 7.0 [1.3-36.6]). There were non-significant associations with total fume exposure (OR = 2.6, 95% CI 0.6-12.2), and exposure index of greater than 10 years (OR=2.8, 95% CI 0.5 15.0). A fall in FEV1 of at least 5% after 15 minutes of work was significantly associated with aluminium exposure (OR=5.8, 95% CI 1.7-20.6). CONCLUSIONS: Nickel exposure from metal inert gas (MIG) and tungsten inert gas (TIG) welding is associated with work-related respiratory symptoms and aluminium exposure from welding is associated with a fall in FEV1 of at least 5 % after 15 minutes of work. PMID- 15133523 TI - Postoperative follow-up strategies for patients after potentially curative surgery for colorectal cancer at Christchurch Hospital. AB - AIM: To describe the follow-up patterns of patients with colorectal cancer (CRC) having had surgery with curative intent. METHODS: A retrospective study was undertaken of follow-up patterns in patients who had undergone 'curative' surgery for colorectal cancer at Christchurch Hospital from 1 January 1996 to 31 December 2000. Patients were identified from three sources: the General Surgical Audit Database (Otago system), the hospital clinical Casemix DRG Database, and the Oncology Service database. Patients were included only if they had surgery with curative intent, within the stated period, and had follow-up at Christchurch Hospital. Data extracted included: patient demographics, details of initial surgery, adjuvant therapies, recurrences, and details of follow-up arrangements (including investigations). RESULTS: Of 893 patients coded as having CRC, 284 patients met the inclusion criteria. Patients were excluded for the following reasons: no operation (64), operation before 1996 (18), palliative surgery (345), previous cancer (55), no cancer (32), died within 30 days of surgery (26), follow up outside of Christchurch region (39), and notes unavailable (30). The median age was 72 (range 28.6-99.9 years). Median follow-up time was 732 days. Most patients (91%) were followed-up by their surgeon. Patients had an average of 2.6 visits to their specialist in the first year of follow-up. Unplanned clinic visits accounted for 8.3% of all clinic visits--resulting in a number of unplanned investigations. During the follow-up period, patients had 112 colonoscopies, 68 CT scans, 8 abdominal ultrasounds, and 7 barium enemas. Recurrence was detected in 58 patients (20.4%); 23 (39.7%) recurrences were detected in the first year of follow-up. Of the 279 patients who had some form of follow-up, 9 asymptomatic patients had recurrent disease (detected as a result of a planned clinic visit) and had a potentially curative procedure for recurrence. CONCLUSIONS: The number of visits per year correlated closely with the earlier findings of Connor et al,4 however the number of investigations carried out was variable and substantially less than had been reported. Follow-up visits have limited value for the detection of asymptomatic potentially curable recurrent disease. PMID- 15133524 TI - Metastatic basal cell carcinoma. PMID- 15133525 TI - Energy settings for mono- and biphasic defibrillation: guideline of the New Zealand Resuscitation Council. PMID- 15133526 TI - Continuing medical education: What for? How? And how much is it worth? PMID- 15133527 TI - The Health Amendment Act allowing access to clinical records. PMID- 15133528 TI - Obesity therapeutics in the shadows: editorial. PMID- 15133529 TI - A population-based cohort study of mortality among users of ibuprofen in Denmark. AB - Using the population-based Pharmacoepidemiologic Prescription Database of North Jutland County, Denmark, we identified 113,538 persons who filled prescriptions for ibuprofen during 1989 through 1995 and determined subsequent mortality through 1996. Standardized mortality ratios [SMRs] for 25 specific causes of death were computed compared with the general population. SMRs were elevated for most causes of death, with an overall SMR of 1.21 (95% confidence interval 1.19 1.24) among persons who filled prescriptions for ibuprofen. There was a nearly threefold increase in the number of deaths from gastrointestinal bleeding within 1 year of ibuprofen prescription but no concomitant increase in hemorrhagic stroke. Elevated SMRs were seen for several cancer types, although the mortality ratios were highest within 1 year of prescription and declined with longer follow up. For colon cancer, SMRs were below 1.0 three or more years after ibuprofen prescription. For hypertensive disease, nonhemorrhagic stroke, and diabetes, we observed slight but significant elevations of the SMRs that persisted beyond the fifth year of follow-up. Our findings indicate a slight increase in overall mortality among persons receiving ibuprofen on prescription. This excess was greatest within the first year, due partially to ibuprofen-related gastrointestinal bleeding but mostly to elevated cancer mortality among ibuprofen users. This temporal pattern is characteristic of an effect of confounding by indication, with ibuprofen being used for pain relief by patients with imminent fatal illnesses such as cancer. The slight excess mortality that persisted beyond the first few years was largely due to elevated death from hypertensive disease and diabetes, which may be explained in part by increased prescription of ibuprofen to patients with long-standing medical problems. PMID- 15133530 TI - Left ventricular ejection fraction change with exercise versus ejection fraction at rest in coronary artery disease: implications for using ejection fraction variations in making therapeutic decisions. AB - Previous studies have differed regarding the prognostic importance of the change (Delta) in left ventricular ejection fraction (LVEF) with exercise among patients with known or suspected coronary artery disease (CAD). Data suggest that these discrepancies may be owing to patient selection, including wide interstudy variations in the range of LVEFrest at study entry; however, the impact of LVEFrest on LVEF exercise response has not been adequately addressed. To test the hypothesis that magnitude and variability in DeltaLVEF are systematically related to LVEFrest, we analyzed data from 2655 patients who underwent rest/exercise radionuclide cineangiography for evaluation of clinically evident CAD, stratified into 5 successive LVEFrest subgroups: <30% (n = 205), 30%-44% (n = 563), 45%-59% (n = 1529), 60%-75% (n = 324), and >75% (n = 34). The standard deviation of DeltaLVEF among patients with LVEFrest <30% was found to be half that among patients in the higher LVEFrest subgroups (P < 0.00001, global). The average magnitude of the rise and fall in LVEF with exercise also varied markedly among LVEFrest subgroups (P < 0.0001, global), being smallest among patients with LVEFrest <30%. These findings may explain differences in predictive accuracy of DeltaLVEF noted among various study populations. Further study is needed to determine whether LVEFrest should be used in selecting exercise-based prognostic descriptors in individual patients. PMID- 15133531 TI - Urinary excretion of rifampicin in the presence of ciprofloxacin. AB - To investigate the effect of the ciprofloxacin on the urinary excretion of the rifampicin in humans, ciprofloxacin and rifampicin were coadministered. Five healthy volunteers between the ages of 20 and 35 years received, on 2 separate occasions (phases 1 and 2) and at weekly intervals, 600 mg rifampicin and 600 mg plus 500 mg ciprofloxacin, respectively, with 350 mL of water. Urinary levels of rifampicin were measured from 1-72 hours later. In phase 1, 15.6% urinary rifampicin was recovered compared with 15.5% urinary rifampicin recovered in the second phase. An increased excretion rate and higher plateau were obtained in ciprofloxacin plus rifampicin treatment. The study indicates that rifampicin may be coadministered with ciprofloxacin to check the development of drug resistance to single-drug therapy by susceptible organisms. PMID- 15133532 TI - Pharmacokinetics, pharmacodynamics, and safety of a lipid-lowering adenosine A1 agonist, RPR749, in healthy subjects. AB - RPR749 and its methylated metabolite are orally active and selective adenosine A(1) agonists that can inhibit lipolysis and lower plasma triglyceride levels in a variety of animal models. RPR749 also appears to lower free fatty acid (FFA) and insulin levels and may have additional lipid-modifying effects. This double blind, single increasing-dose, placebo-controlled, parallel group, randomized study, the first done in humans, evaluated the safety, pharmacokinetics, and pharmacodynamics (effect on FFA) after a single oral dose of up to 200 mg RPR749 or placebo. Six parallel groups of 8 healthy men (6 active and 2 placebo/group) were enrolled in the study. Plasma samples were collected for up to 72 hours post dose. RPR749 and its metabolite RPR772 concentrations were measured by a validated LC/MS/MS method with a minimal quantifiable limit of 1 ng/mL. RPR749 was safe and well tolerated as a single oral dose up to 200 mg. The mean plasma concentrations of RPR749 were approximately 30-fold higher than the mean RPR772 plasma concentrations. The mean terminal half-life (t(1/2)) of RPR749 and RPR772 were similar (approximately 16.4 hours). Mean values for serum insulin, triglycerides, glycerol, and blood glucose remained within normal ranges. Mean FFA concentrations in serum decreased in all treatment groups with the maximal decrease in the 200-mg dose group. In conclusion, RPR749 has the ability to reduce circulating levels of FFA that can be related to plasma RPR749 concentrations and thus possesses pharmacological properties that may be beneficial in treating coronary artery diseases and hyperlipidemia. PMID- 15133533 TI - Bioavailability of metronidazole in rabbits after administration of a rectal suppository. AB - The bioavailability of metronidazole in rabbits was studied using plasma concentration measurements after the administration of the drug in a hydrophilic (glycerogelatin) suppository form. The peak in the plasma concentration time curve occurred about 1 hour after administration, indicating that the rate of absorption is fast and equivalent to that observed in humans after oral administration. There was rapid elimination of the drug, as indicated by a relatively high elimination rate constant and low plasma half-life. The in vitro dissolution profile of the suppositories further confirms rapid absorption of the drug from the suppositories in the rectum. The presence of Tween 80 enhanced the in vitro release of metronidazole, but the presence of a hydrogenated vegetable oil lubricant (Lubritab) caused retardation in the drug release from the suppositories. PMID- 15133534 TI - Cost analysis of fondaparinux versus enoxaparin as venous thromboembolism prophylaxis in hip fracture surgery. AB - Hip fracture patients are at high risk of developing venous thromboembolic events (VTEs) postoperatively. Efficacy and safety data from a clinical trial comparing fondaparinux and enoxaparin as VTE prophylaxis were used to perform a cost assessment of these 2 agents. Incremental cost calculations demonstrate that enoxaparin offers advantages over fondaparinux when dosed for 7 days postoperatively in this patient population. Sensitivity analyses support this finding at the lower extreme; however, fondaparinux provides cost savings at the upper extreme. Cost per death averted and cost per life year gained are similar for these 2 agents in these patients. PMID- 15133535 TI - Clinical experience with perindopril in patients nonresponsive to previous antihypertensive therapy: a large US community trial. AB - A subgroup analysis of a large US community trial was conducted to evaluate the antihypertensive efficacy and safety of perindopril, an angiotensin-converting enzyme inhibitor (ACEI), in 3159 patients who lacked blood pressure (BP) control at entry with previous antihypertensive therapy. Patients received 4 mg perindopril daily for 6 weeks. Based on physicians' assessment of BP response, the patients were then either maintained on 4 mg daily (group 1) or the dose was increased to 8 mg daily (group 2) for an additional 6 weeks. The mean baseline sitting BP was 158.2/92.9 mm Hg. Perindopril monotherapy produced a significant BP decrease from baseline of 11.6/6.5 mm Hg and 14.9/8.4 mm Hg at weeks 6 and 12, respectively. In group 1 patients, the majority of BP decrease occurred at week 6 (17.3/9.5 mm Hg) and was maintained until the end of week 12 (18.2/10.1 mm Hg). In group 2 patients, the BP decrease on the 4-mg dose was modest at week 6 by 5.2/3.1 mm Hg. However, further dose up-titration of perindopril to 8 mg resulted in a clinically significant BP decrease of 11.9/6.8 mm Hg from baseline to week 12. Significant antihypertensive effects of perindopril were also demonstrated in the special patient populations of elderly (>or=65 years), black, isolated systolic hypertension, patients with concomitant cardiovascular diseases, and patients nonresponsive to other ACEI therapy. Overall, BP control (<140/<90 mm Hg) was achieved in 40.0% of patients at week 12. Perindopril was well tolerated with cough and angioedema reported in 8.5% and 0.4% patients, respectively. Physicians assessed therapeutic response to perindopril as satisfactory in 73.8% patients who were nonresponsive to previous antihypertensive therapy. These results suggest that, in a community-based practice, perindopril monotherapy (4-8 mg/d) is an effective and safe therapeutic option in patients nonresponsive to previous antihypertensive therapy. PMID- 15133536 TI - The inhibitory effects of herbal components on CYP2C9 and CYP3A4 catalytic activities in human liver microsomes. AB - Herbal medicines are widely consumed by patients in different clinical settings in the United States and all over the world. In this study, 7 herbal components ginsenosides Rb1, Rb2, Rc, and Rd (from ginseng quercetin) ginkgolides A and B (from ginkgo biloba) were investigated for their inhibitory effects on hepatic CYP2C9 and CYP3A4 catalytic activities in human liver microsomes. Tolbutamide 4 methylhydroxylation and testosterone 6beta-hydroxylation were used as index reactions of CYP2C9 or CYP3A4 catalytic activities, respectively. The metabolites of both reactions were measured by high-performance liquid chromatography and used as indicators of whether enzymes were inhibited or unaffected by these agents. Herbal components were studied at various concentrations (0.1, 1, 10, 100, 200 micromol/L). The herbal compounds investigated were capable of inhibiting CYP2C9 and CYP3A4 catalytic activities, but the potencies differed. Quercetin showed marked inhibitory effects on both tolbutamide 4 methylhydroxylation and testosterone 6beta-hydroxylation with IC(50) values of 35 and 38 micromol/L, respectively. Ginsenoside Rd also had significant inhibitory potency on both CYP2C9- and CYP3A4-mediated index reactions with IC(50) values of 105 and 62 micromol/L, respectively. Ginsenosides Rb1, Rb2, and Rc had limited inhibitory activities on both enzyme reaction systems, whereas the effects of ginkgolides A and B appeared negligible. It is concluded that the components of ginseng and ginkgo biloba screened are capable of inhibiting CYP2C9- and CYP3A4 mediated metabolic reactions. Our findings suggest that quercetin and ginsenoside Rd have the potential to interact with conventional medicines that are metabolized by CYP2C9 and CYP3A4 in vivo. PMID- 15133537 TI - Regulatory toxicology perspectives on the development of botanical drug products in the United States. AB - Toxicological studies constitute an essential part of the effort in developing a botanical supplement into a drug product. The US Food and Drug Administration recently published a draft guidance and established a special botanical review team to assist academic and industry sponsors to manage this and other regulatory considerations related to this unique group of drug products. In this article, the current state of regulatory viewpoints on issues related to requirements and recommendations of various types of nonclinical toxicity studies in support of advanced phases clinical trials and filing a New Drug Application of a botanical are discussed. Topics include nonclinical pharmacology/toxicology view of previous human experience and initial clinical trial, regulatory perspectives on acute toxicity studies, chronic toxicity studies, mutagenicity studies, reproductive toxicity studies, and carcinogenicity studies on botanicals. Certain regulatory review-related issues are also presented. It is anticipated that through a proactive 2-way communication between the Agency and the sponsor, toxicological development of botanical drug product can be significantly facilitated. PMID- 15133538 TI - Sirolimus-eluting coronary stents: novel devices for the management of coronary artery disease. AB - Despite major technological advances in the practice of percutaneous coronary intervention, restenosis of the treated arteries remains a challenge for many interventional cardiologists. Sirolimus is a macrolide antibiotic with potent antifungal, immunosuppressive, and antimitotic activities. Sirolimus inhibits in stent restenosis via 2 major mechanisms of action: by blocking the process of neointimal hyperplasia by inhibiting smooth muscle cell proliferation and by inhibiting inflammatory cell activity. In pivotal clinical trials, the sirolimus eluting stent has demonstrated significant improvements in angiographic and clinical outcomes compared with bare metal stents in patients with de novo lesions in native coronary arteries. Since the systemic exposure of sirolimus in patients who received the drug-eluting stent is minimal, adverse effects resulting from systemic exposure of sirolimus are unlikely to occur. Further studies are needed to determine the safety and effectiveness of sirolimus-eluting stents in patients with more complex coronary artery lesions. In addition, the long-term safety, efficacy, and cost-effectiveness of this novel drug-eluting device will need to be established in ongoing clinical trials. This review article focuses on the pharmacology as well as clinical studies of the sirolimus eluting stent. PMID- 15133539 TI - A case of risperidone-induced hypothermia. AB - Risperidone is one of the second-generation antipsychotics (SGAs). Use of SGAs or so-called atypical antipsychotics is becoming more frequent because they are more efficacious and safer than typical antipsychotics. This is due to their ability to occupy some other receptors as well as dopamine type 2 (D(2)) receptors in the brain. Risperidone has more affinity for serotonin type 2 (5-HT(2)) than for D(2) receptors. This accounts for its better treatment of negative symptoms of schizophrenia and fewer extrapyramidal side effects when compared with typical antipsychotics. Common side effects associated with risperidone include extrapyramidal symptoms, dizziness, nausea, weight gain, sleep disturbance, and sexual dysfunction. We describe here a case of risperidone-induced hypothermia. Body temperature is regulated by the preoptic anterior hypothalamus with involvement of dopamine, serotonin, norepinephrine, and alpha-adrenergic receptors. Experimental data suggest that stimulation of 5-HT(2) and dopamine receptors can increase the body temperature. Additional clinical evidence indicates potent antagonists of 5-HT(2) are more likely to cause hypothermia. Risperidone has higher potency for occupying 5-HT(2) than D(2) receptors. PMID- 15133540 TI - Treatment of sexual disinhibition in dementia: case reports and review of the literature. AB - Behavioral problems are common in dementia patients. They are complex in nature and are often the presenting symptoms of dementia. Its management poses continuous challenge. Gabapentin is an antiepileptic agent that has a role in the management of behavioral problems in demented patients such as agitation and aggression. Its role in treating sexual disinhibition has never been addressed. We describe 3 nursing home residents in 2 dementia units whose sexual disinhibition was effectively treated with gabapentin. PMID- 15133541 TI - Metformin-associated respiratory alkalosis. AB - We present an 84-year-old man with a history of chronic obstructive pulmonary disease, type 2 diabetes, hypertension, glaucoma, and bladder cancer who presented to the emergency department after the police found him disoriented and confused. Metformin therapy began 3 days before, and he denied any overdose or suicidal ideation. Other daily medications included glipizide, fluticasone, prednisone, aspirin, furosemide, insulin, and potassium supplements. In the emergency department, his vital signs were significant for hypertension (168/90), tachycardia (120 bpm), and Kussmaul respirations at 24 breaths per minute. Oxygen saturation was 99% on room air, and a fingerstick glucose was 307 mg/dL. He was disoriented to time and answered questions slowly. Metformin was discontinued, and by day 3, the patient's vital signs and laboratory test results normalized. He has been asymptomatic at subsequent follow-up visits. Metformin-associated lactic acidosis is a well-known phenomenon. Respiratory alkalosis may be an early adverse event induced by metformin prior to the development of lactic acidosis. PMID- 15133542 TI - Vaccine therapy for a patient with androgen-insensitive prostate cancer without evidence of measurable disease on scans. AB - Androgen ablation therapy is the mainstay treatment of patients who have failed local therapy for prostate cancer. PSA is a serum marker commonly used to monitor patients who have undergone primary therapy for evidence of disease progression. However, there currently is no standard therapy for patients once they have demonstrated disease progression. We report a case of a 65-year-old man with castrate testosterone levels but a rising serum PSA level without radiographic evidence of metastasis. After failing prior antiandrogen therapy, he enrolled on a clinical trial comparing a PSA-based vaccine with nilutamide. When his PSA level rose on nilutamide, he commenced treatment with the vaccine therapy. He has continued to have a PSA response with the vaccine treatments with no radiographic evidence disease for >20 months. PMID- 15133543 TI - [Writing English abstracts of high quality in standard English acceptable for publication]. AB - Writing an abstract presents a unique challenge to any author. It requires brevity of the written word, conciseness of thought and accuracy of presentation. A structured abstract is the most important starting point for a well written dental article. Shanghai Journal of Stomatology, a Chinese national journal of oral medicine, oral and maxillofacial surgery, prosthodontics, orthodontics, oral pathology, endodontics, oral radiology, etc, has been included and indexed in Chemical Abstracts, Index Medicus and MEDLINE. This peer reviewed refereed journal demands Chinese and English abstracts of high quality for all manuscripts. Each manuscript should provide qualified abstracts for citation and indexing, which are written in standard English using the correct terms, sentences and grammar. Structured abstracts about 500 characters are required for clinical articles, clinical reports, basic scientific articles; while indicative abstracts (summary) are mandatory to short communications and case reports as well as review articles. Generally, the abstracts should be written in the past tense and passive voice (in the third person), with exception of conclusion(s), which can be in the primary tense and active voice. The structured abstracts are consisted of PURPOSE (S), METHODS, CONCLUSION (S), and CONCLUSION (S), which should offer the important information on the number of the subjects, methods of grouping and statistics, database attained, P values, etc. The conclusion (s) must be detailed rather than conceptive. This editorial introduces the ins and outs of writing an English abstracts acceptable for publication. PMID- 15133544 TI - [Immunodetections and analysis of peripheral blood for patients with oral cancer undergoing operation]. AB - PURPOSE: To investigate the immunological function for patients with oral cancer undergoing operation. METHODS: Peripheral blood was taken from patients with oral cancer before and after operation. Proportions of CD3(+) total T, CD4(+) Th, CD8(+) Tc/Ts, CD19(+) total B, CD16(+)/CD56(+)NK were detected by flow cytometry (FCM). Detections of serum tumor necrosis factor alpha (TNF-alpha) and soluble interleukin-2 receptor (sIL-2R) were performed with enzyme-linked immunosorbent assay(ELISA). Paired t test was used for statistical analysis. RESULTS: The proportions of CD8(+)Tc/Ts, CD19(+) total B and CD4(+)/CD8(+) increased significantly after surgery. Post operative proportion of CD16(+)/CD56(+)NK decreased more significantly. The concentration of TNF-alpha was less than that before operation(P<0.05). CONCLUSIONS: Surgery could change immunological function and alleviate immunosuppression of oral cancer patients. Detections for immune conditions of oral cancer patients might have important clinical values in prognosis evaluation and guiding treatment, especially immunotherapy, after operation. PMID- 15133545 TI - [Treatment and prevention of cerebrospinal fluid leakage after craniofacial surgery for tumors involving the skull base]. AB - PURPOSE: To evaluate the treatment and prevention of cerebrospinal fluid (CSF) leakage following removal of the skull base tumors with craniofacial approach. METHODS: A retrospective review was undertaken to analyze the clinical data of 14 cases suffering from CSF leakage after craniofacial surgery for tumors involving the cranial base. RESULTS: Ten (71.4%) case cured between 3 days and 32 days by conservative treatment including lumbar drainage. Three of four patients who needed operation had no CSF leakage after first attempt. One discontinuous leakage was closed after a second attempt. Among all cases,five(35.7%) patients developed meningitis,and one case died because of serious intracranial infection and respiration failure. CONCLUSION: Continuous lumbar cerebrospinal fluid drainage is a safe and effective treatment of CSF leakage following craniofacial surgery. However,If conservative treatment fails to arrest CSF leakage,surgical therapy is recommend with multilayer reconstruction for dural defect so as to avoid meningitis. The vital procedure for prevention of CSF leakage is watertight seal of dura mater, reconstruction of bone defect and obliteration of dead space after craniofacial surgery. PMID- 15133546 TI - [Clinical observation and treatment of failing flaps in oral and maxillofacial surgery: a report of 44 cases]. AB - PURPOSE: For the purpose of enhancing the successful transplanting rate of free flaps and reviewing the clinical characters, a retrospective study was undertaken to analyze the causes as well as clinical characters of the failing flap in oral and maxillofacial regions. METHODS: A total of 832 free revascularized flaps were performed in our department from August 1997 to April 2003. The data including the time to occurence, causes, clinical characters and the managements of flap crises were reviewed and then analyzed. RESULTS: Forty-four flaps developed vascular crisis (5.29%), most of them were caused by venous occlusion (38/44). Thirty-eight in forty-four flaps crises occurred within 72 hours (86.36%). 32 flaps crises were explored with 21 flaps successfully salvaged; the success rate was as high as 65.63%. CONCLUSIONS: For the highest successful transplanting rate of free flaps, the management of the local factors plays an important role. Most of the failing flaps happened within three days after operation. The surgical exploration should be done as soon as possible, if the crises flap happened within six hours. PMID- 15133547 TI - [The diagnostic value of magnetic resonance sialography for chronic obstructive parotitis]. AB - PURPOSE: To evaluate the diagnostic value of magnetic resonance sialography (MRS) for chronic obstructive parotitis (COP). METHODS: 18 patients with COP underwent both conventional sialography and MRI sialography. A new magnetic resonance technique was applied. In addition to the usually performed T1 and T2 cross sectional sequence, a heavy T2-weighted sequence (TR=4000 msec,TE=250 msec)was performed that allowed depiction of the fluid-filled parotid duct system. The MRI sialographic findings were compared with that of conventional sialography. The overall accuracy of diagnosis and ductal stenosis were assessed. RESULTS: The main duct of the parotid gland as well as primary branching ducts could be reliably depicted. Compared with the conventional sialography, the diagnostic accuracy of MRS was 94.4%(17/18) and stenosis diagnosis reached 100%(17/17). CONCLUSIONS: Initial experience indicates that magnetic resonance sialography can be applied successfully to investigate the duct system of the parotid gland. It is completely noninvasive and a promising alternative to radiographic sialography. PMID- 15133548 TI - [The influence of occlusion on the incidence of bruxism in 779 children in Shanghai]. AB - PURPOSE: To explore the influence of occlusion on the incidence of bruxism in children. METHODS: The incidence of bruxism in children was investigated from the questionnaire of 779 children's parents. The occlusion of these children was checked by professionals. The influence of the occlusion on the incidence of bruxism in children was analysed. Chi-square test was used for statistical analysis. RESULTS: The occlusion of children with bruxism and children without bruxism had no significant difference (P>0.05). The incidence of bruxism did not increase in the children with mixed dentition(P>0.05). CONCLUSION: The occlusion does not affect the incidence of bruxism in children. The mixed dentition does not increase the incidence of bruxism in children. PMID- 15133549 TI - [Effect of intraoperative radiation therapy on enzyme histochemistry of expression muscle of guinea pigs after injury of facial nerve]. AB - PURPOSE: To explore the mechanism of facial nerve recovery after radiotherapy by simulating surgical treatment of parotid gland carcinoma with reserving facial nerve and studying the ability of aerobic metabolism, transmission of neurotransmitter, variation of ultrastructure of motor end and mitochondrion after radiotherapy during operation. METHODS: Animal models of treatment group(15Gy) and lethal dose group(17Gy) were established. Succinic dehydrogenase(SDH) and acetylcholine lipase(AchE) of orbicularis oris were measured after radiotherapy. Student's t test was used for statistical analysis. RESULTS: The activity of SDH and AchE declined after radiotherapy. There was significant difference between treatment group and lethal dose group. One month later, there was a significant improvement in the activity of SDH and AchE in treatment group, while that of lethal dose group continued to decline. CONCLUSION: After radiotherapy during parotid gland surgery, the activity of SDH and AchE, transmission of neurotransmitter, and the ability of aerobic metabolism decreased. The ultrastructure of motor end and mitochondrion was destroyed. The variation returned to preoperative levels in treatment group, while did not in lethal dose group. PMID- 15133550 TI - [Investigation of primary cell culture for dental pulp: comparing of coronal pulp and root pulp]. AB - PURPOSE: To compare and establish an ideal method for the culture of human primary dental coronal and root pulp cells. METHODS: The whole dental pulp tissues were obtained from healthy and young human teeth extracted for orthodontic purposes. The cells in coronal and root pulp were separately cultured by the tissue explant and tissue explant-collagenase digestion(0.1% type I collagenase) methods. By evaluating the attachment efficiency,viability and the successfulness of the cultured cells, the optimal primary cell culture process of human dental coronal and root pulp was determined. RESULTS: The tissue explant culture had more successfulness,more viability and less attachment efficiency than that of tissue explant-collagenase digestion method in the culture of human coronal and root dental pulp cells. CONCLUSIONS: Tissue explant method was superior to tissue explant-collagenase digestion method for the culturing of human dental coronal and root pulp cells. The ideal method for dental coronal and root pulp cells was established in this study. PMID- 15133551 TI - [The characteristics of cultured dental pulp cells and the localization of dental pulp stem cells]. AB - PURPOSE: This study was aimed to investigate the localization of dental pulp stem cells (DPSCs) by comparing the characteristics of cultured dental pulp cells in coronal and root pulp. METHODS: Human dental coronal and root pulp cells were cultured in tissue-explant method, the cell culture successfulness, attachment efficiency, cell viality, morphology, proliferation pattern, and the mineralization ability were observed, the localization of DPSCs was investigated in the functional respect of DPSCs. RESULTS: The human dental root pulp cells have more culture successfulness,more attachment efficiency,more cell viality, more primary characteristics, and stronger induced mineralization ability than that of coronal pulp cells. Root and coronal pulp cells showed same proliferation patterns. CONCLUSIONS: DPSCs may exist in both dental root and coronal pulp, and the density of DPSCs in the root pulp may be higher than the coronal pulp. PMID- 15133552 TI - [The establishment of the individual model libraries for stomatognathic system]. AB - PURPOSE: To investigate the method of establishing the individual model libraries for stomatognathic system. METHODS: Based on the normal 3D FE models, including the upper jaw, lower jaw and whole dentitions, with the help of the ANSYS5.5, divided, trimmed or re-built these models, created the abnormal stomatognathic system. RESULTS: Established three model libraries in common use, including the alveolar-absorbed-teeth, loosed-teeth, and alveolar-reduced-area, 308 models in all. CONCLUSION: The established models have a satisfied geometric configuration, it will be more suitable for the research and clinic works if uses the individual FE models. PMID- 15133553 TI - [The establishment of 3D-FE model libraries for the elements of the RPD]. AB - PURPOSE: To establish the 3D-FE model libraries for the elements of the RPD. METHODS: Based on the 3D geometric model libraries of the RPD and normal stomatognathic system, using the software, such as Powershape,Pro/E and Ansys5.5, transformed data, trimmed models, or rebuilt it, established many kinds of FE model elements, then combine these elements in the whole model, connected and trimmed, at last created several kinds of FE model libraries. RESULTS: This study established eight kinds of FE model libraries, including the occlusal rests, clasps, bases, artificial teeth, and connectors, etc, nearly two hundreds FE model elements. CONCLUSION: The established models have a satisfied geometric configuration; these elements combined well in the whole model, and can be easily call-in and edit. PMID- 15133554 TI - [The establishment of 3D-FE model libraries for the elements of FPD]. AB - PURPOSE: To establish the 3D-FE model libraries for the elements of the FPD. METHODS: Based on the 3-D geometric model libraries of the normal stomatognathic system, using the software, such as Powershape,Pro/E and Ansys5.5, the models of abutments,retainers and pontics were created according to the models of natural crowns. The models of connectors were established according to the sutures between the crowns. RESULTS: This study has established the FE model libraries for the elements of the FPD, including the retainers,pontics,and connectors, three kinds of libraries and 82 elements in all. CONCLUSION: The established models have a satisfied geometric configuration; the analysis models can be easily established and edited. PMID- 15133555 TI - [Effect of stress distribution of support tissues using hinge extracoronal attachment prostheses]. AB - PURPOSE: The purpose is to discuss the clinical indications of hinge extracoronal attachment. METHODS: In the experiment, the stress distribution in the periodontal support tissue of the abutment was analyzed under different numbers of abutments by EFA. RESULTS: There was no significant effect on the stress distribution in the supporting tissue according to the different numbers of abutments at the central occlusion, and there was an evenly stress distribution in the abutment and alveolar membrane of the area around the missing teeth. CONCLUSIONS: When using the attachment prostheses to restore the type of the second molar distal-extension for the patient, we can use one teeth as abutment. The attachment is suitable for the cases with better condition of the alveolar membrane without high demand of the abutment teeth of the patient. PMID- 15133556 TI - [Giant branchial cyst with infection in newborn infant: case report]. AB - Branchial cyst is a rarely seen inheritent cyst commonly located in the lateral face and neck.A giant branchial cyst with infection in a newborn infant was reported in this paper. Surgical enucleate of the cyst was performed at 17 days of the infant with good result. The patient has been followed up for 2 years without recurrence. PMID- 15133557 TI - [Study on odontal & periodontal tissue of guinea pigs, dogs and monkeys]. AB - PURPOSE: To evaluate the different application of different experimental animals in studies of periodontal diseases. METHODS: The odontal and the periodontal tissues of guinea pigs, dogs and monkeys were studied by gross specimen, X ray and histological observation. RESULTS: The most suitable sites of guinea pigs for periodontal study is incisor just because of their small oral fissure, but the keratinized epithelium of gingival sulcus may affect their inflammatory reaction. As the occurring of spaces from the 3rd to 5th tooth of dogs, artificial intrabony pocket can be made, and their thick root canals are suitable for pulp and periapical study. The monkeys are the ideal experimental animals due to their similarities to human, but it is too expensive to be widely used. CONCLUSION: Each kind of animal has their own characteristics. The key is to select experimental animal according to the objective of our different studies. PMID- 15133558 TI - [Cloning of human amelogenin gene encoding mature peptide]. AB - PURPOSE: The purpose of this study was to clone human amelogenin gene encoding mature protein, which provides a basis for expressing the recombinant human amelogenin in Escherichia coli. in the future. METHODS: In this study, total RNA was extracted from the dental germ of a legally aborted embryo by Trizol. Using RT-PCR technique we obtained synthesis of cDNA from the total RNA, and the desired DNA products were conducted with PCR from cDNA. The segment was inserted into expression vector PQE30 and the interesting plasmid was transformed into Escherichia coli. host DH5alpha. The double-stranded DNA of positive clone was analyzed by PCR, restriction endonuclease mapping and DNA sequence analysis. RESULTS: The sequence analysis of recombinant plasmid showed that the human amelogenin encoding mature protein was inserted into vector PQE30 accurately. CONCLUSIONS: We conducted human amelogenin encoding mature protein from dental germ of a legally aborted embryo and got the recombinant plasmid which may express amelogenin gene for further research. PMID- 15133559 TI - [Morphologic characteristics of anterior crossbite in early permanent dentition reviewed by superimposition on the template]. AB - PURPOSE: To establish a computer-processing module to first, eliminate the factors which mislead the diagnosis of anterior crossbite and second, superimpose the individual tracing on the template to review the patient's morphologic characteristics. METHODS: First, the module would process the following steps:1.to create three templates; 2.to move the functional-shift-mandible posteriorly; 3.to standardize the potential of vertical dimension;4.to select a suitable template for superimposition; 5.to standardize the sizes of individual graph and template. Second, the module would process the five-step superimpositions: 1. X axis was superimposed and S were registered to measure the distances between Ptm points and Ar points on X axis; 2. X axis was paralleled and Ptm were registered to measure the distance between A points on X axis; 3. X axis was paralleled and A points were registered to measure the difference of anterior area between the mandibular graphs; 4.mandibular planes were superimposed and Po was registered to show the mandibular form variation; 5. Ar Gn lines were superimposed and Ar was registered to measure the difference between the mandibular graphs. RESULTS: The module which could complete these steps was produced. With its help, the individual graph could be diagnosed with the position of maxilla and mandible, the amounts of maxillary discrepancy and jaws' dysplasia, the mandibular form and size variation. CONCLUSIONS: The module could eliminate the factors which mislead the diagnosis and review the morphologic characteristics of anterior crossbite in early permanent dentition. PMID- 15133560 TI - [A review of statistical analysis methods in measurement data]. AB - Measurement data, which is obtained by measuring the size of numeric value, is consisted of numerical variables. In clinic research, the collected data covers quite of numerical variables. Composed of those numerical variables, the measurement data has to be correspondingly analyzed in the special statistical methods in the light of their design methods. The statistical methods of measurement data include descriptive statistics and inferential statistics. Descriptive statistics is mainly used to describe the distribution laws and the characters/features of the data. For instance, when measuring the central tendency and dispersion tendency of the data, methods such as calculating the mean and the standard deviation can be considered. Inferential statistics is mainly employed to estimate the confidence limit such as estimating the 95% confidence interval and to have the hypothesis testing in the example of having t test and analysis of variance. By highlighting the specific statistical methods of measurement data, this review intends to help the clinicians and researchers select correct statistical methods in accordance with measurement data. PMID- 15133561 TI - [Research advances and applications of the methods to repair mandible defect]. AB - Mandible defects can be caused by many diseases, most commonly occurring in cranio-maxillofacial regions. With the development of orthopaedics, biomaterial and cell biology in these years, great improvements have been acquired in repair of mandible defects. This paper systematically reviewed the relevant research advances and applications, and showed the applied prospect of bone tissue engineering in this field. PMID- 15133562 TI - [Advances in investigation of calcified root canal therapy]. AB - Calcified root canal occurs often during root canal therapy. It's important to understand various methods to deal with calcified root canal, which may increase the successful rate. From anatomy, radiography, chemical root canal preparation, ultrasonic root canal preparation and endodontic microscopic system etc., this paper reviews the present research about the treatment techniques for calcified root canal. PMID- 15133563 TI - [Surgical laser treatment for base cell carcinoma of maxillofacial region]. AB - PURPOSE: Base cell carcinoma is a common skin malignancy involved in the facial area. Present surgical removal, cryosurgery and laser evaporation are the effective modalities. Following the development of laser technology, we used fine focused carbon dioxide laser surgery to treat basal cell carcinoma of the maxillofacial region. METHODS: Six cases of base cell carcinoma were included in this study. Tumors were removed by CO2 laser scalpel under local anesthesia. Intra-operative laser biopsy and post-operative follow up was carried out. RESULTS: six cases had radical result with a follow up of two years post operatively with satisfactory contour. CONCLUSION: Laser scalpel excision can get radical result for basal cell carcinoma with advantages of safety, simple and less bleeding. It is recommended to be used. PMID- 15133564 TI - [Clinical results of 46 patients with jaw fractures but delayed management]. AB - 46 cases with jaw fractures but delayed management were reviewed, the time, causes of delayed treatment and outcomes were analyzed retrospectively. The delayed time to treatment was between 2 to 12 days. The causes of delay were multiple. All the patients underwent surgical treatment including open reduction and internal fixation. The results showed that 6 patients were treated 2 to 5 days after injury with primary healing. 31 patients were treated 5 to 10 days after injury, 27 were with primary healing. 9 patients were treated 10 to 12 days after injury with primary healing. PMID- 15133565 TI - [Comparison of the effects of three analgesic therapies on odontalgia caused by pulpitis in molars]. AB - PURPOSE: To compare the effects of analgesic therapies on odontalgia caused by pulpitis in molars by three different methods. METHODS: 173 molars were diagnosed as irreversible pulpitis, and randomized block designed in three groups. Under block or local anesthesia: 46 cases of the first group, leaving teeth on open drainage, after 1 day, sealing devitalized material (arsenic), arranged next appointment 2 weeks later. 52 molars of the second group, enlarging exposure site, drainage several minutes only, sealed devitalized material, re-examined 2 weeks later. 75 cases of the third group, removing roof of pulp chamber, undergoing pulpectomy directly, re-examined 1 week later. Adopting VAS(vital analogue scale) to analyze the pulp receptivity of three different methods and to evaluate the analgesic effects by complete analgesia, effective analgesia, and no response. The data was analysed using chi-square test. RESULTS: The rate of complete analgesia was 50.0%, 63.46% and 76.0%, respectively. There was a significant difference in complete analgesia among the three methods. The analgesic effect of the third group (pulpectomy) was significantly higher than that of the first group (P<0.01).Pulpectomy group had impossibility of adverse effects caused by using devitalizing material (arsenic). CONCLUSIONS: It is worthy to adopt pulpectomy extensively to relieve the pain of molars caused by pulpitis clinically. The course of treatment of the third group was significantly shorter than other groups, and the simultaneous symptoms occurred rarely. PMID- 15133566 TI - [The effect of initial therapy on periodontal status and saccharified Hb (HbAIc) of patients with type II diabetes mellitus]. AB - PURPOSE: To investigate the effect of periodontal initial therapy on the level of glycated hemoglobin(HbAIc) and periodontal status in non-insulin dependent diabetes mellitus(NIDDM)patients with periodontitis. METHODS: 33 cases with NIDDM periodontitis were included in the study. The periodontal therapy included oral hygiene instruction,ultrasonic scaling and subgingival scaling. The level of glycosylated hemoglobin and periodontal status before treatment and four weeks after treatment were determined and compared. RESULTS: The percentage of bleeding on probing and probing depth were significantly reduced in all patients after periodontal therapy. The glycosylated hemoglobin level was significantly decreased in patients with advanced periodontitis,while patients with moderate periodontitis showed no changes following therapy. CONCLUSION: The result of periodontal therapy in the diabetic patients was satisfied in short time. It can reduce the level of glycated hemoglobin. PMID- 15133567 TI - [Discussion of surgical laser manipulation of venous malformations in the parotid gland region]. AB - PURPOSE: This study is carried to explore the surgical manipulation and complications of Nd:YAG laser coagulation for venous malformations in parotid gland region. METHODS: The study included 84 cases of venous malformations in the parotid gland region, who underwent laser therapy after surgical approach three years ago. The results were evaluated by CT or MRI. Postoperative facial nerve function and intraoperative bleeding were analyzed. RESULTS: Five cases had temporary facial nerve paralysis, which recovered spontaneously two months postoperatively. One case developed permanent nerve paralysis, which accounted for 1.19% in this group. The mean amount of blood loss was 200 ml. Four cases received blood transfusion. CONCLUSION: Careful surgical dissection and correct laser irradiation is the key to successful treatment, which could preserve facial nerve from injury and decrease bleeding. PMID- 15133568 TI - [The use of BD-018 medical natural absorbable surgical suture in oral and maxillofacial surgery]. AB - The oral and maxillofacial cutaneous or/and oral mucous wound of 20 cases were sutured with BD-018 medical natural absorbable surgical suture. Fifteen cases with class I wound and five cases with class II wound healed well after operation. BD-018 medical natural absorbable surgical suture adapts to oral and maxillofacial cutaneous or/and oral mucous wound suturing. PMID- 15133569 TI - [Correction of linguoclination or buccoclination of the mandibular molar with simple semi-fixed lingual arch]. AB - PURPOSE: Introducing an effective corrective therapy for linguoclination or buccoclination of the mandibular molars. METHODS: Using a simple semi-fixed lingual arch combined with the edgewise technology to correct 8 cases with linguoclination or buccoclination of the mandibular molars. RESULTS: Using the simple semi-fixed lingual arch can correct the malposition of mandibular molar's linguoclination or buccolination quickly and successfully. CONCLUSION: The simple semi-fixed lingual arch can fix properly the buccolingual position of the mandibular molar and can shorten the course of the fixed orthodontic therapy. In addition,this method exerts less impact on the contour of the anterior mandibular arch. PMID- 15133570 TI - [Using fenestration and decompression technique to treat enormous odontogenic keratocyst]. AB - 3 cases of enormous odontogenic keratocyst were treated with the technique of fenestration and decompression other than removal of jaws. After a follow-up period of two years, radiographic assessment showed that the image of keratocyst disappeared basically, neogenetic bone had been found and the defect of jaws had been restored. The teeth had no dysfunction, the lower lip had no numbness and the keratocyst epithelium was modulated histologically to mucosa after decompression. Fenestration and decompression are satisfied conservative approaches to treat enormous keratocyst which usually is treated by removal of jaws and extraction of teeth. PMID- 15133583 TI - Anterior cruciate ligament reconstruction with doubled semitendinosus and gracilis tendon graft in rugby players. AB - The purpose of this study was to evaluate prospectively the results of anterior cruciate ligament (ACL) reconstruction with doubled hamstring tendon graft in a selected group of 18 rugby players. The graft was fixed with a transcondylar screw (Transfix) on the femur, and with an absorbable interference screw and a metallic staple on the tibia. All the patients followed the same rehabilitation program. Return to sports activities was allowed after 6 months. Follow-up was 2 years in all cases. The athletic level of the patients was rated according to the Tegner scoring system. Clinical results were evaluated using the International Knee Documentation Committee (IKDC) scoring system. Furthermore, an instrumented evaluation of the anterior laxity with a KT-1000 arthrometer, and an isokinetic evaluation were performed 6 and 24 months after surgery. The Tegner mean score at follow-up (8.2) was similar to that prior to injury (8.3). IKDC overall results were normal in ten cases (55.6%), nearly normal in six cases (33.3%), and abnormal in two cases (11.1%). Side-to-side difference of anterior laxity measured with KT-1000 at 6 and 24 months did not show an impairment of knee stability with time. Isokinetic evaluation showed a significant improvement on peak torque both in extension and flexion on comparison between 6- and 24-month measurements. The results reported in this study showed that the use of doubled hamstring tendon graft for ACL reconstruction in athletes that were at risk for high-energy traumas to the knees, such as rugby players, gave normal or nearly normal results in about 90% of the cases. Recovery of muscle strength was almost complete 2 years after surgery, and there was no impairment of knee stability with time. PMID- 15133584 TI - The incidence of motocross injuries: a 12-year investigation. AB - Off-road motorcycling is one of the most popular sports activities practiced by millions of people in the world but little has been written on motocross traumatology and its prevention. This paper aims to evaluate motocross injuries in terms of injury ratio, location, causes, and possible prevention in a series of competitions organized by Motorcyclistic Federations over a 12-year period. We retrospectively evaluated 1,500 accidents with 1,870 rider injuries out of a group of 15,870 athletes participating in European off-road competition from 1980 to 1991. Data were collected from race medical reports, insurance declarations and follow-up forms filled up by riders involved in accidents. We then classified the type and location of the injury, modality of the accident, the protective gear used and the recovery of the riders. We compared our data to lesions noted in motorcycle road races using the chi-square test and the z-test. The overall incidence of motocross injuries in our study was 94.5 per thousand, while stadium cross competitions had a 150 per thousand rate and outdoor motocross a rate of 76 per thousand representing a risk of accident of 22.72 per thousand hours of riding. Among the total of 1,870 injuries, 1076 were bruises; 27.9% of these were in the upper extremities, 26.9% on the lower, 21.2% on the trunk, and 16% on the face. There were 450 fractures recorded, 50.9% in the upper extremities, 38% in the lower, and the rest were on the spine, chest, and skull. The 26 spine fractures (5.8%) produced permanent neurologic sequelae in eight patients. Ligamentous lesions accounted for 344 cases with 206 (59.9%) occurring in the lower extremities especially on the knee (42.4%). Head trauma was noted in 86 cases (5.7% of accidents) producing coma in 3%, and loss of consciousness in 14%. Limb involvement for all types of injuries were more frequent on the left side (60%). Motocross is a high-risk sport: our study revealed the most common modalities and types of lesions sustained by the riders. Despite the reduction of some injuries by better protective gears, the occurrence of knee sprain, and wrist and clavicular fractures are still high. Furthermore, the high number of spine lesions with subsequent neurologic deficit noted in indoor races raises doubts about the safety of these events. PMID- 15133589 TI - Schizophrenia and employment - a review. AB - BACKGROUND: Little is known about the extent to which work contributes to the recovery of people with schizophrenia. There is increasing interest in the subject because of new service models and the economic cost of unemployment in people with severe mental illness. METHODS: A literature search was carried out with the aim of investigating: a). employment rates in schizophrenia and first episode psychosis and the extent to which they have changed over time; b). the barriers to work; c). the factors associated with being employed among people with schizophrenia; and d). whether employment influences other outcomes in schizophrenia. RESULTS: There are wide variations in reported employment rates in schizophrenia. Most recent European studies report rates between 10 % and 20%, while the rate in the US is less clear. There is a higher level of employment among first-episode patients. The employment rate in schizophrenia appears to have declined over the last 50 years in the UK. Barriers to getting employment include stigma,discrimination, fear of loss of benefits and a lack of appropriate professional help. The most consistent predictor of employment is previous work history. Working is correlated with positive outcomes in social functioning, symptom levels, quality of life and self esteem, but a clear causal relationship has not been established. CONCLUSIONS: Very low employment rates are not intrinsic to schizophrenia, but appear to reflect an interplay between the social and economic pressures that patients face, the labour market and psychological and social barriers to working. PMID- 15133590 TI - Migration and schizophrenia: the challenges for European psychiatry and implications for the future. AB - BACKGROUND: The last decade of the twentieth century has seen an unprecedented increase in the number of reports in the psychiatric literature documenting increased rates of psychotic illness among migrants in a range of European countries. In countries where high rates of immigration have been long-standing such as Britain and the Netherlands, these increased rates have also been seen in the second generation of migrants. This has impacted on psychiatry significantly with regard to the aetiology, diagnosis, and treatment of schizophrenia. METHOD: We reviewed the literature to summarise the available evidence about the phenomenon across the European countries where these findings have been reported. Comparisons of the findings between countries were highlighted to establish their impact on psychiatry and to identify areas and implications for future research. RESULTS: The history of this kind of research is longest in Britain and has established increased risk for non-white migrants, with Caribbean and African patients being especially vulnerable. Caribbean migrants to the Netherlands have also been found to be at increased risk, but they are predominantly of Indo Caribbean ethnicity. In the other European countries, East and West African migrants have been implicated in some countries, while European migrants have been implicated in other countries. Social inequalities, family fragmentation and urbanicity seem to be the main hypotheses proposed for these increased rates, though, in some countries where asylum seekers and refugees form the largest group of migrants, the stress of the migratory process itself may be implicated. These may all interact with genetic vulnerability and substance abuse. DISCUSSION: Ethnicity and differences in dominant language emerge as major structural references in this new epistemology of psychosis and both the causes and the effects on psychopathology may be filtered through an experience of social disadvantage in an urban environment. PMID- 15133591 TI - An RCT of early intervention in psychosis: Croydon Outreach and Assertive Support Team (COAST). AB - BACKGROUND: Despite considerable interest in early intervention in psychosis, the evidence base for its effectiveness is sparse. We aimed to evaluate a new service in South London, UK, Croydon Outreach and Assertive Support Team (COAST) using a randomised controlled trial (RCT) during its first year. METHOD: Referrals were taken from local adult community mental health teams of those with documented first service contact in the last 5 years and a diagnosis of any functional psychosis. Those who consented (N = 59) were randomised to COAST or treatment as usual (TAU). COAST offered a range of interventions, including optimum atypical medication, psychological interventions (individual cognitive behavioural therapy and family intervention if appropriate) and a range of vocational and welfare help according to need. Whole team training was used to be able to offer these kinds of interventions. RESULTS: Outcomes were evaluated at baseline, 6 months and 9 months on a range of standardised clinical and social measures. Overall both COAST and TAU clients improved over time, but there were no significant improvements for COAST clients; a lack of significant results in the time x treatment interaction. There was a trend for COAST carers' quality of life to increase. Bed days were also less in COAST, but not significantly so. CONCLUSIONS: The lack of clearly demonstrated improvements for COAST is consistent with the published literature so far. The fact that both groups improved in symptoms and functioning over the year suggests that while access to early intervention is helpful, community adult mental health teams should aim to offer high quality input at any stage of psychosis in order to meet client and carer needs. PMID- 15133592 TI - Is the excess risk of psychosis-like experiences in urban areas attributable to altered cognitive development? AB - BACKGROUND: Rates of psychotic disorder and related attenuated psychotic experiences are higher in urban areas. We examined to what degree differences between urban and rural areas could be attributed to differences in cognitive development. METHOD: Scores on the nine subscales of the schizotypal personality questionnaire (SPQ) as well as IQ and specific neuropsychological functions of memory and attention were assessed in a representative sample of 943 young army conscripts from the 49 counties of Greece. RESULTS: Young men from urban areas had higher scores on the SPQ subscale Odd beliefs/magical thinking (OR = 1.99, 95% CI: 1.42, 2.78), but lower scores on Excessive social anxiety (OR = 0.63, 95 % CI: 0.49, 0.81) and No close friends (OR = 0.42, 95% CI: 0.29, 0.62). Adjustment for demographic factors, IQ and specific neuropsychological functions did not change the results. When the lower scores on Excessive social anxiety and No close friends were taken into account, the differences on the Odd beliefs/magical thinking subscale became even more pronounced (OR = 2.33, 95% CI: 1.56, 3.49). CONCLUSIONS: Young men from urban areas are socially more competent, but display higher levels of positive psychotic experiences, which are not mediated by lower IQ or higher levels of neuropsychological impairment. PMID- 15133594 TI - Patients with a first episode of schizophrenia spectrum psychosis and their pathways to psychiatric hospital care in South Germany. AB - BACKGROUND: Several first-episode studies of schizophrenia suggest that many patients experience psychotic symptoms for a long time before receiving appropriate treatment. To reduce the time of untreated psychosis, it is necessary to know the patients' pathways to psychiatric care. This study was designed to examine patients' help-seeking contacts and the delays on their pathways to psychiatric care in Germany. METHOD: Sixty-six patients with first episode of schizophrenia spectrum psychosis were assessed by the Interview for the Retrospective Assessment of the Onset of Schizophrenia (IRAOS) and were interviewed about their help-seeking contacts before psychiatric admission. RESULTS: In contrast to other findings of long duration of untreated psychosis (DUP), 53% of our patients were admitted after 8 weeks (median) of untreated positive symptoms, although the mean value of 71 weeks corresponds well with the results of other studies. There were important differences in DUP depending on which kind of statistical parameter (median or mean) was used. In contrast to studies from other countries, only 18% of our patients had their first contact with a general practitioner. However, this was the fastest way to psychiatric admission. No differences were found between patients with short (< 1 year) and long (> 1 year) DUP in the duration of time from the first help-seeking contact up to admission. CONCLUSION: In Germany, a large number of mental health professionals in private practice or different services of psychosocial contact facilities exist in every region and general practitioners are not so important as a link to psychiatric care, although they seem to be functioning well if it is necessary. Therefore, programs designed to reduce the delay of treatment should focus less on general practitioners than on other health services. PMID- 15133593 TI - Evaluation of familial influences on the course and severity of schizophrenia among US and Indian cases. AB - BACKGROUND: Prior studies suggest familial (possibly genetic) influences on the course of schizophrenia. AIMS: The aim of this study was to compare familial influences on the course and severity of schizophrenia in two independent samples. METHOD: Thirteen selected measures were compared among affected sibling pairs (ASPs) from Pittsburgh, USA and New Delhi, India (48 US pairs, 53 Indian pairs). For each ASP proband, an unrelated patient was selected randomly from a suitable pool of cases ascertained in the same study (Sibpair proband-comparison case or S-C pairs). Correlations between these pairs were compared. RESULTS: The correlations varied by item and by site. Significant correlations for longitudinal course and pattern of severity were noted among the ASPs from USA, but did not remain significant following corrections for multiple comparisons. Comparisons between the correlations for ASPs and the S-C pairs, used to estimate familial effects, yielded trends for the ASP correlations to be numerically larger than the S-C correlations in both samples. Separate cross-site comparisons revealed several significant differences with regard to several demographic and clinical variables. The possible impact of the cross-site variations on the observed ASP correlations is discussed. CONCLUSIONS: Though familial factors did not appear to have a significant impact on course/severity using this novel design, the suggestive trends need to be examined in larger samples. PMID- 15133595 TI - Needs in outpatients with schizophrenia, assessed by the patients themselves and their parents and staff. AB - BACKGROUND: There has been a change in psychiatric care from a hospital-oriented care system to an outpatient-centred system, which has underlined the importance of support alternatives. METHODS: The sample was drawn from outpatients with schizophrenia at an outpatient clinic in 2001. We used structured interviews when interviewing outpatients, parents and staff. The interview with the patient included the patient's needs, global function, clinical global impression and insight. Both parents and staff were interviewed about the patient's needs. RESULTS: The mean value of GAF was 56 +/- 10, CGI 4 +/- 1 and 89% of the patients had full insight into their illness. The patients rated the total score of the severity of needs at a mean of 7 +/- 4, while the parents' and staff's rating was 9 +/- 5. The needs ranking between patients and parents and patients and staff showed a correlation of rho = 0.65 (p < 0.01) and parents and staff rho =0.95 (p < 0.01). The parents rated more problems involving physical health and money than the patients. CONCLUSION: When planning mental health in the future, it is important to assess the views of the patients, the parents and the staff from a multiple perspective. PMID- 15133596 TI - Needs for care of chronic schizophrenic patients in long-term community treatment. AB - BACKGROUND: Specific problems of long-term community care of chronic schizophrenic patients are an under-researched area interesting for the provision of regional mental health care. METHODS: This study focuses on a 4 1/2-year prospective assessment of normative needs for care in a cohort (initially N = 115) living in the Dresden care region (Germany). At six time-points, normative needs for care were assessed with the Needs for Care Assessment (NFCAS). RESULTS: The total number of problems did not change significantly over the study period. The average number of met needs was lower at the longer-term follow-up assessments,due particularly to a decrease in the social section. This trend is also demonstrated for the average number of unmet needs. In contrast, the mean number of "unmeetable needs" increased. Consistently, 70-80% of the patients exhibit problems in positive psychotic and negative symptoms, household affairs and recreational activities. Communication, occupation and recreational activities constitute a trio of social needs not met for nearly one-third of the patients disabled in these respects. Logistic analyses of regression could not identify a predictive model for the total needs development within the 4 1/2-year community treatment. CONCLUSION: The rather stable pattern of needs for care seems to define clear long-lasting tasks for community mental health services. For chronic schizophrenic patients, services should especially focus on social skills training and psychoeducational approaches. Due to a wide range of possible factors of influence, however, planning long-term context-dependent processes of care in the community lacks a clear evidence base. PMID- 15133597 TI - Quality of life in patients with schizophrenia--comparison of self-report and proxy assessments. AB - BACKGROUND: While Quality Of Life (QOL) in subjects suffering from schizophrenia has been studied using a variety of generic or specific instruments, only very few studies have analyzed the agreement between patients and proxy ratings on patients' QOL. METHODS: We administered the World Health Organization Quality of Life assessment instrument (WHOQOL-100) to 292 patients and the Quality of Life for Proxies (QOL-P) to their proxies, respectively; the QOL-P is a 30-item instrument derived from the WHOQOL-100 and adapted for administration to a key informant. RESULTS: Agreement between patients and proxies on the four main QOL areas was highest for the physical area (intraclass correlation coefficient, ICC = 0.41) and lowest for the psychological area (ICC = 0.29). In line with the results of other studies comparing patients' and proxies' ratings, proxies generally underestimated patients' physical and psychological QOL. Moreover, the agreement between patients' and proxies' ratings was consistently higher across all QOL areas when the proxy was a relative compared to a non-relative proxy. CONCLUSIONS: The agreement between patients and proxies in QOL assessment is modest, but it is relatively higher when observable aspects of QOL are rated and when the proxy who makes the evaluation is a family member who has closer contacts with the patient. In order to obtain a comprehensive picture of patients' QOL, it would be advisable to compare patients' ratings with the assessments made by close informants. PMID- 15133598 TI - Teachers' knowledge, beliefs and attitudes concerning schizophrenia- a cross cultural approach in Japan and Taiwan. AB - BACKGROUND: Mental health literacy of the general public is essential for the effective promotion of society's mental health. However, there has been no investigation of the general public's mental health literacy with Japanese and Taiwanese socio-cultural backgrounds. METHODS: A total of 129 Japanese and 150 Taiwanese elementary school teachers were surveyed about knowledge, beliefs and attitudes concerning schizophrenia by means of a questionnaire with a vignette describing a case of the disease. Identification of the case, cause of the disease, coping behavior for the case, and perception of stigmatizing and supporting attitudes by parents and neighbors of the case were investigated. RESULTS: As a common finding with the studies in Western countries, only small percentages of the Japanese and Taiwanese respondents were able to make a correct identification. A further common finding was the emphasis on psychosocial factors as a cause of schizophrenia, as was the rejection of psychotropic medication, although future study is required to determine to what extent the respondents know about therapeutic procedures utilized by psychiatrists. Significantly stronger stigma perception was shown in the Japanese respondents than in the Taiwanese, which may be attributable to the high institutionalization rate in Japan. CONCLUSIONS: Japanese and Taiwanese teachers' knowledge, beliefs and attitudes regarding schizophrenia were similar to those found in the general public in Western societies. Although the present study is limited in sampling and the components of the mental health literacy investigated, several working hypotheses have been extracted from it to be tested in future investigations on the Japanese and Taiwanese and other Asian general public's mental health literacy. PMID- 15133599 TI - Perception of patients' unpredictability and beliefs on the causes and consequences of schizophrenia- a community survey. AB - BACKGROUND: The belief that mental disorders involve a high risk of unpredictable behaviours is a factor which influences negatively the social acceptance of the mentally ill. In this paper, we compare the beliefs about the causes and psychosocial consequences of schizophrenia expressed by 536 respondents who had the firm conviction that patients with schizophrenia are unpredictable and by 457 respondents who firmly believed that they are not. METHODS: The survey was conducted in 30 Italian geographic areas, randomly selected taking into account their location and population density. The data were collected by the Questionnaire about Opinions on Mental Illness (QO). RESULTS: Respondents who believed that patients with schizophrenia are unpredictable reported more frequently factors such as use of alcohol and drugs and frequenting bad company as being involved in the development of the disorder. In addition, this group showed more restrictive opinions about patients' civil and affective rights. Low education was found to be significantly associated with perception of "unpredictability" in schizophrenia. CONCLUSIONS: These results suggest the need to: a). inform the general public on the main clinical characteristics of schizophrenia and on the risk of unpredictable behaviours in the acute phases of this mental disorder; b). carry out sensitisation campaigns against discrimination toward people with schizophrenia emphasising successful experiences of social integration. PMID- 15133606 TI - Identification of quantitative trait loci for wood and fibre properties in two full-sib properties of Eucalyptus globulus. AB - Regions of the genome influencing wood and fibre traits in Eucalyptus globulus Labill. have been identified in two full-sib pedigrees that share a common male parent. The first pedigree, cross A, contains 148 progeny, and the second pedigree, cross B, contains 135 progeny. Subsets of progeny of these two controlled crosses were planted at seven sites throughout Australia in 1990. Wood cores were taken at 0.9 m above ground in 1997, and wood and fibre traits were analysed for each individual. Three quantitative trait loci (QTL) affecting wood density, one QTL affecting pulp yield and one QTL affecting microfibril angle have been located in both pedigrees, using single-factor analysis of variance. Other QTLs affecting these traits, as well as fibre length and cellulose content, were located in cross A only. PMID- 15133611 TI - Spinal and bulbar muscular atrophy: ligand-dependent pathogenesis and therapeutic perspectives. AB - Spinal and bulbar muscular atrophy (SBMA) is a late-onset motor neuron disease characterized by proximal muscle atrophy, weakness, contraction fasciculations, and bulbar involvement. SBMA exclusively affects males, while females are usually asymptomatic. The molecular basis of SBMA is the expansion of a trinucleotide CAG repeat, which encodes the polyglutamine (polyQ) tract in the first exon of the androgen receptor (AR) gene. The histopathological hallmark is the presence of nuclear inclusions containing mutant truncated ARs with expanded polyQ tracts in the residual motor neurons in the brainstem and spinal cord, as well as in some other visceral organs. The AR ligand, testosterone, accelerates AR dissociation from heat shock proteins and thus its nuclear translocation. Ligand-dependent nuclear accumulation of mutant ARs has been implicated in the pathogenesis of SBMA. Transgenic mice carrying the full-length human AR gene with an expanded polyQ tract demonstrate neuromuscular phenotypes, which are profound in males. Their SBMA-like phenotypes are rescued by castration, and aggravated by testosterone administration. Leuprorelin, an LHRH agonist that reduces testosterone release from the testis, inhibits nuclear accumulation of mutant ARs, resulting in the rescue of motor dysfunction in the male transgenic mice. However, flutamide, an androgen antagonist promoting nuclear translocation of the AR, yielded no therapeutic effect. The degradation and cleavage of the AR protein are also influenced by the ligand, contributing to the pathogenesis. Testosterone thus appears to be the key molecule in the pathogenesis of SBMA, as well as main therapeutic target of this disease. PMID- 15133614 TI - [Modern differential therapy with diuretics]. AB - Diuretics block different electrolyte transporters in renal tubular cells. Their predominant action is inhibition of renal sodium chloride reabsorption, however, and achievement of a negative body sodium balance is the principal goal of diuretic therapy in patients with hypertension and edema. Several classes of diuretics can be distinguished with respect to the sites of sodium reabsorption along the nephron, but loop diuretics and distal-tubular diuretics (incl. thiazides) are the most widely used. The latter have a less potent natriuretic effect than loop diuretics, but their long duration of action predispose them for treatment of patients with uncomplicated hypertension. In conditions of gross edema, e.g. heart and/or renal failure, distal-tubular diuretics lose their efficacy and must be replaced by or combined with loop diuretics ("sequential nephron blockade"). Aldosterone antagonists are unique among diuretics because they improve survival in patients with heart failure independently of their effect on sodium metabolism. PMID- 15133612 TI - [35-year old male with fever and round mass of the liver]. AB - A 35 year old male suffered from fever till 39 degrees C and malaise since 6 months; infectious or neoplastic causes have been ruled out. Ultrasonography revealed a hypoechoic lesion of 75 mm diameter in the liver. The histologic examination of the needle biopsy showed an inflammatory process, and the final diagnosis was inflammatory pseudotumor. EBV specific LMP-1 protein was detected within the tumor immunohistochemically. A liver segment resection was performed, and the patient has been well and without fever since then for 18 months. Inflammatory pseudotumor is a rare but important differential diagnosis in cases of hypoechoic liver lesions associated by fever, if there is no abscess or malignancy. The diagnosis must be confirmed histologically, because imaging techniques cannot reliably classify the lesion. PMID- 15133615 TI - [HNO-relevant changes in the current TNM-classification of malignant tumours]. PMID- 15133616 TI - [Emergencies in allergology]. AB - The clinical presentation of allergological emergencies is heterogeneous. The most important conditions are anaphylaxis (which is frequent) and Stevens-Johnson syndrome or toxic epidermal necrolysis (which are rare). In the acute phase it is most important to recognize the exogenous elicitation of the condition as soon as possible, otherwise severe sequelae, even death of the patient, may occur. Management is based on immediate elimination of possible causative agents and symptomatic therapeutic measures. Furthermore, allergological diagnostics are necessary without delay to identify what has elicited the reaction. This information makes possible long-term treatment in order to avoid further hypersensitivity reactions. PMID- 15133617 TI - [The dermatologic consultation]. AB - BACKGROUND: Dermatology and venereology, as a cross-sectional discipline, plays an important role in the interdisciplinary care of inpatients. Nonetheless, only a few studies on dermatological consultations have been performed. We reviewed the dermatological consulting service at the University Hospital of the Martin Luther-University, Halle, Germany. PATIENTS AND METHODS: All consultations performed in 2001 and 2002 were retrospectively assessed. In addition, the time spent in consulting activities was prospectively recorded over a period of six months (January-June 2003). RESULTS: A total of 2390 consultations were evaluated. Most of the consultations (42.8%) were made on internal medicine wards, followed by pediatrics (11.7%), neurology (9.9%) and cardiothoracic surgery (5.7%). Infectious skin diseases accounted for the most frequent diagnosis at 24.4%. The time lapse between request and performance of the consultation averaged 1.1 days. Patients older than 40 years needed dermatological consultation more frequently than younger patients. The duration of consultation averaged 23 minutes (time to site, patient examination, documentation). Thus, the man-hours required for the consulting service is 33.6 per month. CONCLUSION: The study emphasizes that dermatological consultation services make an important contribution to the care of inpatients. In light of increasing specialization in medicine, the consulting service guarantees interdisciplinary treatment of the patients and is thus an important quality parameter for inpatient care. PMID- 15133618 TI - [Special emergencies in dermatology]. AB - Emergency cases in dermatology are rare but potentially life-threatening conditions. They comprise a broad spectrum of diseases which require immediate hospitalization or even intensive care. A rapid diagnosis and appropriate therapy is necessary since some of these emergency cases are associated with a high mortality and with severe disabling complications. Typical examples are: the bacterial infections Waterhouse-Friderichsen syndrome, necrotizing erysipelas and necrotizing fasciitis, as well as staphylococcal and streptococcal toxic shock syndromes. Another special emergency situation is venomous snake bites. PMID- 15133619 TI - The role of stress in the growth of a multicell spheroid. AB - Rather recent experimental results demonstrate the non-negligible role of mechanical stress in the growth of a multicell spheroid. In this paper we discuss a theoretical framework for volumetric growth suitable for modeling the growth of soft tissues exhibiting the properties of a solid. After a proper kinematic decomposition, balance equations for mass, momentum and energy are discussed together with constitutive relationships. The mathematical model is then applied to avascular tumor growth. We show by numerical simulation that, under assumption of spherical symmetry, the mathematical model is able to reproduce the experimental data with a satisfying qualitative agreement. PMID- 15133620 TI - Malthusian parameters, reproductive values and change under selection in self fertilizing age-structured populations. AB - A population, reproducing wholly by selfing, is assumed to be observed at times 0,1,.... Individuals between x-1 and x units of age at time t are said to be in age class x at that time. The rate of increase in the long run of individuals of type A(iA)(j) is denoted by m(ij)+1= m(ji)+1. For each genotype there is also a set of reproductive values, corresponding to all age classes and genotypes of individuals having descendants of that genotype. Then, if the number of individuals of each sort of ancestor is multiplied by its reproductive value and the products are summed, the result is the total value, which is V(ij)( t) for genotype A(iA)(j). Then V(ij)( t+1)- V(ij)( t) is equal to m(ij) V(ij)( t), where m(ij) is the Malthusian parameter for A(iA)(j). Furthermore, if the mean and variance at time t of the m(ij)'s, weighted by their corresponding reproductive values, are respectively ( t) and Sigma(m)(2)( t), then m( t+1)-m( t)=Sigma(m)(2)( t)/(1+m( t)). PMID- 15133621 TI - Permanence of single-species stage-structured models. AB - In this paper, we consider population survival by using single-species stage structured models. As a criterion of population survival, we employ the mathematical notation of permanence. Permanence of stage-structured models has already been studied by Cushing (1998). We generalize his result of permanence, and obtain a condition which guarantees that population survives. The condition is applicable to a wide class of stage-structured models. In particular, we apply our results to the Neubert-Caswell model, which is a typical stage-structured model, and obtain a condition for population survival of the model. PMID- 15133622 TI - A framework for modelling and analysing conspecific brood parasitism. AB - Recently several papers that model parasitic egg-laying by birds in the nests of others of their own species have been published. Whilst these papers are concerned with answering different questions, they approach the problem in a similar way and have a lot of common features. In this paper a framework is developed which unifies these models, in the sense that they all become special cases of a more general model. This is useful for two main reasons; firstly in order to aid clarity, in that the assumptions and conclusions of each of the models are easier to compare. Secondly it provides a base for further similar models to start from. The basic assumptions for this framework are outlined and a method for finding the ESSs of such models is introduced. Some mathematical results for the general, and more specific, models are considered and their implications discussed. In addition we explore the biological consequences of the results that we have obtained and suggest possible questions which could be investigated using models within or very closely related to our framework. PMID- 15133623 TI - Optimal HIV treatment by maximising immune response. AB - We present an optimal control model of drug treatment of the human immunodeficiency virus (HIV). Our model is based upon ordinary differential equations that describe the interaction between HIV and the specific immune response as measured by levels of natural killer cells. We establish stability results for the model. We approach the treatment problem by posing it as an optimal control problem in which we maximise the benefit based on levels of healthy CD4+ T cells and immune response cells, less the systemic cost of chemotherapy. We completely characterise the optimal control and compute a numerical solution of the optimality system via analytic continuation. PMID- 15133624 TI - Minimal entropy probability paths between genome families. AB - We develop a metric for probability distributions with applications to biological sequence analysis. Our distance metric is obtained by minimizing a functional defined on the class of paths over probability measures on N categories. The underlying mathematical theory is connected to a constrained problem in the calculus of variations. The solution presented is a numerical solution, which approximates the true solution in a set of cases called rich paths where none of the components of the path is zero. The functional to be minimized is motivated by entropy considerations, reflecting the idea that nature might efficiently carry out mutations of genome sequences in such a way that the increase in entropy involved in transformation is as small as possible. We characterize sequences by frequency profiles or probability vectors, in the case of DNA where N is 4 and the components of the probability vector are the frequency of occurrence of each of the bases A, C, G and T. Given two probability vectors a and b, we define a distance function based as the infimum of path integrals of the entropy function H( p) over all admissible paths p(t), 0 < or = t< or =1, with p(t) a probability vector such that p(0)=a and p(1)=b. If the probability paths p(t) are parameterized as y(s) in terms of arc length s and the optimal path is smooth with arc length L, then smooth and "rich" optimal probability paths may be numerically estimated by a hybrid method of iterating Newton's method on solutions of a two point boundary value problem, with unknown distance L between the abscissas, for the Euler-Lagrange equations resulting from a multiplier rule for the constrained optimization problem together with linear regression to improve the arc length estimate L. Matlab code for these numerical methods is provided which works only for "rich" optimal probability vectors. These methods motivate a definition of an elementary distance function which is easier and faster to calculate, works on non-rich vectors, does not involve variational theory and does not involve differential equations, but is a better approximation of the minimal entropy path distance than the distance //b-a//(2). We compute minimal entropy distance matrices for examples of DNA myostatin genes and amino-acid sequences across several species. Output tree dendograms for our minimal entropy metric are compared with dendograms based on BLAST and BLAST identity scores. PMID- 15133625 TI - In vitro assessment of nucleoside analogs in multiple myeloma. AB - PURPOSE: To identify nucleoside analogs that may be effective for multiple myeloma (MM), we tested fludarabine, clofarabine, arabinosylguanine, cytarabine, troxacitabine, and gemcitabine in MM cell lines. METHODS: We employed biologic and biochemical assays in MM cell lines to evaluate the clinical potential of these nucleoside analogs. RESULTS: Among these purine and pyrimidine nucleoside analogs, fludarabine, clofarabine and gemcitabine were the most potent. MM cell lines, resistant to commonly used chemotherapeutic agents for this disease, were more sensitive to gemcitabine with an IC50 in the nanomolar range. The greater cytotoxicity of gemcitabine in MM cells was consistent with greater accumulation of gemcitabine triphosphate, the major cytotoxic metabolite of this drug. MM.1S cells accumulated >100 microM gemcitabine triphosphate but accumulated <20 microM of the other analogs as the respective triphosphates. In addition incubation with gemcitabine resulted in inhibition of DNA synthesis. Incubation with 25, 50 or 100 nM gemcitabine resulted in a dose- and time-dependent increase in the cell population with a subG1 DNA content indicative of apoptosis. CONCLUSIONS: These results suggest that gemcitabine is a potent nucleoside analog in MM cell lines including cell types resistant to other chemotherapeutic agents. The greater activity of gemcitabine compared to other analogs seems to be due to favorable metabolism of this agent. PMID- 15133626 TI - Synergistic cytotoxicity of the ribonucleotide reductase inhibitor didox (3,4 dihydroxy-benzohydroxamic acid) and the alkylating agent carmustine (BCNU) in 9L rat gliosarcoma cells and DAOY human medulloblastoma cells. AB - PURPOSE: Ribonucleotide reductase (RR) is the rate-limiting enzyme of de novo DNA synthesis and has been shown to be upregulated linked with proliferation and malignant transformation. It was therefore identified as an excellent target for antitumor therapy. In the present study we investigated the biochemical and cytotoxic effects of didox, an inhibitor of RR, as a single agent and in combination with BCNU, an alkylating anticancer drug, in 9L rat gliosarcoma cells and DAOY human medulloblastoma cells. METHODS: The effect of didox on the intracellular concentrations of deoxynucleosidetriphosphates (dNTPs) was studied in 9L cells. Pool sizes were determined by HPLC. In addition, the cytotoxic effects of didox and BCNU as single drugs and in equimolar combination were tested in 9L and in DAOY cells. Combination effects were determined according to the equation of Chou and Talalay. The expression of DNA repair-related genes was determined after exposure of 9L cells to BCNU, didox and a combination of the two compounds, using a cDNA array. RESULTS: Incubation of 9L cells with 30 microM didox for 24 h significantly decreased the intracellular concentrations of the DNA precursors dCTP (61% of control) and dGTP (17% of control), and significantly increased the concentration of dATP (155% of control). This dNTP imbalance compromised DNA synthesis and repair and might therefore have been, at least in part, responsible for the highly synergistic cytotoxic effects seen when BCNU was used simultaneously with didox in 9L and in DAOY cells. With almost all combinations tested, highly synergistic effects were seen, as indicated by combination indices of <1 according to the equation of Chou and Talalay. In 9L cells, BCNU upregulated the expression of DNA repair-associated genes, whereas coincubation of the cells with didox reduced overexpression of some of these repair-related genes. CONCLUSION: A combination of BCNU and didox was proven to act in a synergistic manner in two cell lines, 9L rat gliosarcoma and DAOY human medulloblastoma cells. Further in vivo tests using these two compounds systemically and/or locally at the tumor site might be warranted. PMID- 15133627 TI - Pharmacokinetically guided dosing of carboplatin in paediatric cancer patients with bilateral nephrectomy. AB - An approach to carboplatin dosing in children with bilateral nephrectomy using a renal function-based dosing formula with a glomerular filtration rate of zero was investigated in the current study. Carboplatin exposure was determined in a total of nine courses of chemotherapy in four patients with Wilms' tumour. Carboplatin exposures following initial dosing were less than 50% of the defined target area under the plasma concentration-time curve (AUC) in all four patients studied, with actual AUC values of between 31% and 45% of the target exposures. The use of real-time pharmacokinetic monitoring to guide dosing within a course of carboplatin treatment resulted in exposures within 15% of the target AUC in all patients. Using this information to guide dosing on additional courses of treatment in the same patient resulted in consistent exposures without the need for further monitoring or dose adjustment. These results indicate that real-time pharmacokinetic monitoring of carboplatin treatment plays a key role in ensuring that an appropriate exposure to carboplatin is achieved in children with bilateral nephrectomy. Carboplatin dosing based on patient body weight, or use of a fixed dose of carboplatin, would both be predicted to result in individual patients receiving unsatisfactory drug exposures. Further studies are warranted to further elucidate the relationship between non-renal clearance of carboplatin and patient body weight in this and other patient subpopulations where there remains concern about the optimal way to use this anticancer drug. PMID- 15133628 TI - Dexamethasone as a probe for docetaxel clearance. AB - PURPOSE: A pilot study was conducted in 23 patients in order to assess the correlation between docetaxel clearance (CL) and pharmacokinetics of dexamethasone. Dexamethasone is mainly 6-beta hydroxylated by CYP3A4, and is regularly used as standard docetaxel premedication. Genotyping of known functional single nucleotide polymorphism (SNP) of CYP3A5 (G22893A) and mdr-1 (G2677T, G2677A, and C3435T) have been performed in order to tentatively correlate genotype with docetaxel and dexamethasone pharmacokinetics. PATIENTS AND METHODS: To be eligible for this study, patients were required to have a solid malignancy for which docetaxel was indicated. A population pharmacokinetic approach was used to determine individual pharmacokinetic parameters of both docetaxel and dexamethasone by Bayesian analysis, and to screen relationships between docetaxel CL and patients' demographic, phenotype and genotype covariates. RESULTS: Three different pharmacokinetic parameters of dexamethasone were significantly correlated with docetaxel CL: dexamethasone plasma clearance (DPC) that ranged between 7.7 and 27.2 l/h, urinary amount of 6beta hydroxydexamethasone, and the ratio between urinary amount of 6beta hydroxydexamethasone and unchanged dexamethasone. The best covariate model was docetaxel CL (l/h) = 356 x fu(alpha1-AG) x (1-0.17 x HPMT)(1+0.126 x DPC) where fu(alpha1-AG) is the unbound plasma fraction of docetaxel calculated from alpha1 acid glycoprotein plasma level, and HPMT is hepatic metastasis coded as 1 if present or 0 if absent. No significant difference in docetaxel CL was observed between the several genotypes. CONCLUSIONS: Dexamethasone may be used as a probe to predict docetaxel clearances, hence reducing interindividual variability. PMID- 15133629 TI - Pulmonary infiltrates in patients with haematologic malignancies: transbronchial lung biopsy increases the diagnostic yield with respect to neoplastic infiltrates and toxic pneumonitis. AB - We retrospectively evaluated 107 fiberoptic bronchoscopies with and without transbronchial lung biopsy (TBLB) in 98 consecutive patients with haematologic malignancies and pulmonary infiltrates. Bronchoalveolar lavage (BAL) was performed in 45 and BAL plus TBLB in 62 procedures. There was no procedure related severe haemorrhage, pneumothorax or death. Infectious aetiology was identified in 26 of 107 (24%), toxic pneumonitis in 17 of 107 (16%) and neoplastic infiltration in 9 of 107 (8.5%) episodes. Combined BAL and TBLB was significantly superior to BAL alone with respect to the diagnosis of neoplastic infiltrates (p=0.008) and toxic pneumonitis (p<0.001) and should therefore be included in the diagnostic work-up of this patient cohort. PMID- 15133630 TI - Functional characterization of T lymphocytes derived from patients with acute myelogenous leukemia and chemotherapy-induced leukopenia. AB - T-cell-targeting immunotherapy is now considered in acute myelogenous leukemia (AML). Immunotherapy seems most effective for patients with a low AML cell burden, and a possible strategy is therefore to administer immunotherapy early after intensive chemotherapy when patients have a low leukemia cell burden and severe treatment-induced cytopenia. To further investigate this possible therapeutic approach we used a whole blood assay to characterize the proliferative responsiveness (3H-thymidine incorporation) of circulating T cells from AML patients with severe treatment-induced leukopenia, i.e., peripheral blood leukocyte counts < 0.5x10(9)/l. This assay will reflect both quantitative and qualitative differences. Responses were compared for 17 AML patients, 6 patients with acute lymphoblastic leukemia (ALL), and a group of 21 healthy controls. Most circulating leukocytes in the AML patients were T lymphocytes, whereas B lymphocytes and monocytes usually constituted < 10%. Anti-CD3 stimulated proliferation was significantly lower for AML patients compared with healthy controls. However, proliferation in response to anti-CD3 + anti-CD28 did not differ for AML patients and healthy controls, an observation suggesting that T cells from AML patients have an increased responsiveness in the presence of optimal costimulation that compensates for the quantitative T-cell defect. In contrast, the responses were significantly lower for ALL than for AML patients. We conclude that the remaining T-cell population in AML patients with severe chemotherapy-induced cytopenia show an increased proliferative responsiveness and may represent a therapeutic target when antileukemic immunotherapy is tried in combination with intensive chemotherapy. PMID- 15133631 TI - Proteasomal cleavage does not determine immunogenicity of gp100-derived peptides gp100 209-217 and gp100 209-217T210M. AB - Immune responses against tumor-associated antigens rely on efficient epitope presentation. The melanoma-associated antigen (Ag) gp100 contains HLA-A*0201 ligands that are characterized by low to medium binding affinity, among which gp100(209-217) is the most prominent (Kawakami et al., J Immunol 154:3961-3968, 1995). While this epitope is a natural T-cell target, it primes with low efficiency T-cell responses during immunization. A modified gp100 epitope, gp100(209-217T210M), that contains a Thr to Met substitution at position 2 of the antigenic nonamer is characterized by high binding affinity for HLA-A*0201 and elicits strong and clinically effective T-cell responses. This higher affinity is believed to represent the sole reason for enhanced immunogenicity. Contrasting with this observation is the unpredictable relationship between affinity and immunogenicity observed in other antigen systems. In addition, we noted a striking difference between the capability of endogenously processed gp100(209 217) and gp100(209-217T210M) to induce T-cell responses in an in vitro model. Therefore, we questioned whether factors other than HLA-affinity might play a role in determining the immunogenicity of these epitopes. In the present study, we evaluated the in vitro proteasomal cleavages of 23meric precursor peptides encompassing the native sequence (gp100(201-223)) or the modified sequence (gp100(201-223T210M)). Here we show that the standard proteasome liberates the C termini of both antigenic peptides but not the N-termini. Quantitative analysis of the digestion products revealed that more of the fragments displaying the final C-termini were produced from the wild-type precursor. However, a stronger TCR engagement was observed when fractions of digested gp100(201-223T210M) were used to activate an HLA-A*0201-expressing target T-cell clone. This difference was also found using separately produced, synthetic nonamers. In conclusion, the high binding affinity of gp100(209-217T210M) seems to compensate for possible differences in proteasomal cleavage at the biological level. Since the final antigenic nonamer is not directly produced by the proteasome, additional further factors may influence the antigenic peptide availability, such as post proteasomal processing and intracellular peptide transport. PMID- 15133632 TI - Strategies for immune therapy: Wurzburg, Germany, 29 February-3 March 2004. PMID- 15133633 TI - (201)Tl-SPECT in low-grade gliomas: diagnostic accuracy in differential diagnosis between tumour recurrence and radionecrosis. AB - PURPOSE: The aim of this work was to describe the usefulness of a simple (201)Tl single-photon emission computed tomography (SPECT) technique in the differential diagnosis between tumour recurrence and radionecrosis during the follow-up of patients treated for low-grade gliomas. METHODS: The study population comprised 84 patients treated for low-grade gliomas who showed suspicion of tumour recurrence during their follow-up. All patients were examined by neuro-anatomical imaging procedures (CT, MRI) and (201)Tl-SPECT. (201)Tl-SPECT images were assessed by visual analysis based only on the information on the prescription form and by estimation of the uptake index (ratio of mean counts in the lesion to those in the contralateral mirror area). Examiners were blinded to the results of other tests. RESULTS: Under these conditions, the neuro-anatomical procedures yielded 26.2% inconclusive reports, with a global diagnostic accuracy of 0.61, a sensitivity of 0.63 and a specificity of 0.59. The global diagnostic accuracy for (201)Tl-SPECT was 0.83, with a sensitivity of 0.88 and a specificity of 0.76. Diagnostic pitfalls were observed in regions with physiological (201)Tl uptake, i.e. the posterior cranial fossa, diencephalon, lateral ventricles and cavernous and longitudinal venous sinuses. An uptake index cut-off value of 1.25 showed a sensitivity of 0.90 and specificity of 0.80 for detection of tumour activity. CONCLUSION: (201)Tl-SPECT has adequate diagnostic accuracy to be part of routine algorithms in the follow-up of patients with low-grade glioma suspected of tumour recurrence, as an alternative to neuro-anatomical procedures and not solely as a complementary test. PMID- 15133634 TI - FDG PET and other imaging modalities in the primary diagnosis of suspicious breast lesions. AB - Mammography is the primary imaging modality for screening of breast cancer and evaluation of breast lesions (T staging). Ultrasonography is an adjunctive tool for mammographically suspicious lesions, in patients with mastopathy and as guidance for reliable histological diagnosis with percutaneous biopsy. Dynamic enhanced magnetic resonance mammography (MRM) has a high sensitivity for the detection of breast cancer, but also a high false positive diagnosis rate. In the literature, MRM is reported to have a sensitivity of 86-96%, a specificity of 64 91%, an accuracy of 79-93%, a positive predictive value (PPV) of 77-92% and a negative predictive value (NPV) of 75-94%. In unclarified cases, metabolic imaging using fluorine-18 fluorodeoxyglucose positron emission tomography (FDG PET) can be performed. In the literature, FDG PET is reported to have a sensitivity of 64-96%, a specificity of 73-100%, an accuracy of 70-97%, a PPV of 81-100% and an NPV of 52-89%. Furthermore, PET or PET/CT using FDG has an important role in the assessment of N and M staging of breast cancer, the prediction of tumour response in patients with locally advanced breast cancer receiving neoadjuvant chemotherapy, and the differentiation of scar and cancer recurrence. Other functional radionuclide-based diagnostic tools, such as scintimammography with sestamibi, peptide scintigraphy or immunoscintigraphy, have a lower accuracy than FDG PET and, therefore, are appropriate only for exceptional indications. PMID- 15133635 TI - FDG-PET for axillary lymph node staging in primary breast cancer. AB - Management of the axilla in patients with operable breast cancer is still one of the most controversial areas in clinical oncology. The best procedure to examine the lymph nodes is still standard axillary lymph node dissection; nevertheless, the morbidity associated with this procedure is well known. Based on these considerations, it is important for progress in the treatment of operable breast cancer that strategies are found that permit a less invasive method of axillary sampling which does not impair the patient's quality of life. The technique of sentinel lymph node (SLN) biopsy has recently been proposed for this purpose, with very important results. SLN has now become routine practice in the surgical management of breast cancer, and in many institutions patients with a negative SLN biopsy are spared axillary dissection, while those with a positive SLN biopsy are submitted to axillary node dissection. The good accuracy of SLN biopsy represents a significant advance in the management of primary breast cancer; however, false negative axillary results can occur in a variable percentage of patients, and the contribution of the SLN procedure to the detection of metastases in the internal mammary and supraclavicular lymph nodes is not clear. Among the recently developed imaging modalities, positron emission tomography (PET) with (18)F-fluorodeoxyglucose (FDG) has in particular been applied to the study of lymph node metastases in cancer patients. Several clinical studies have been carried out to evaluate the accuracy of PET in the axillary staging of operable primary breast cancer. These studies have sometimes provided conflicting results, either supporting the possibility of using FDG-PET to select patients who need axillary dissection or questioning whether FDG-PET can accurately assess the axillary status in primary breast cancer. All the limitations and the advantages of FDG-PET are discussed in this paper, by examining the performance of scanner technology and the possible causes of the false negative results. In the experience of the authors, comparing FDG-PET with SLN biopsy in the same series of patients, the results seem to indicate that the lower sensitivity of PET is restricted to micrometastases. Of course, this limitation of PET has to be analysed in relation to the importance of such small axillary metastases for the outcome of patients with breast cancer. The added value offered by PET in breast cancer staging in comparison with intraoperative detection of the sentinel node lies in the fact that FDG-PET is a non-invasive procedure that allows, within a single examination, the biological characterisation of breast cancer and viewing of the entire body. PMID- 15133636 TI - PET/CT and breast cancer. AB - During the past decade, the application of positron emission tomography with [(18)F]fluoro-2-deoxy-D-glucose (FDG-PET) has remarkably improved the management of cancer patients. Nevertheless, the clinical interpretation of FDG-PET scan can be difficult for two main reasons: (1) anatomical localisation of FDG uptake is not easy, (2) normal physiological accumulation of FDG can be misinterpreted as a pathologic area. It has been demonstrated that the visual correlation of PET with morphological procedures, such as computed tomography or magnetic resonance imaging, can improve the accuracy of PET alone. However, the time interval between the two scans, the time employed by the operator and difficulties in co registering imaging of the abdomen and pelvis make the co-registration of separately obtained images clinically difficult. A novel combined PET/CT system has been built that improves the capacity to correctly localise and interpret FDG uptake. To date only a few studies have been conducted on the potential role of PET/CT in the management of breast cancer patients, but the better performance of this technique compared with PET alone should also be relevant for breast cancer application. In this review, we evaluate the possible impact on breast cancer diagnosis of PET/CT compared with PET alone, with respect to disease re-staging, treatment monitoring, preoperative staging and primary diagnosis. In addition, the possible role of PET/CT for radiotherapy planning is evaluated. PMID- 15133637 TI - Combined use of fine-needle aspiration biopsy, MIBI scans and frozen section biopsy offers the best diagnostic accuracy in the assessment of the hypofunctioning solitary thyroid nodule. AB - PURPOSE: The probability of malignancy is increased in hypofunctioning solitary thyroid nodules (HFNs). Fine-needle aspiration biopsy (FNA), (99m)Tc methoxyisobutylisonitrile (MIBI) and frozen section biopsy (FS) have limited independent diagnostic accuracy for the differential diagnosis of HFNs. The goal of this study was to assess the accuracy of the three independent diagnostic methods in distinguishing between benign and malignant disease. METHODS: A total of 130 patients with an HFN on the (99m)Tc-pertechnetate scan were included in this study. FNA, MIBI scans, FS, thyroidectomy and histological analysis of surgical specimens for final diagnosis were performed in all patients. RESULTS: Of the 130 patients, 80 (61.54%) had benign lesions and 50 (38.46%), malignant lesions. FNA was diagnostic in 78/130 (60%) patients and non-diagnostic in 52/130 (40%) patients. None of the patients with a negative MIBI scan had a final histological diagnosis of malignancy, and MIBI scans were negative in 38.46% of patients with non-diagnostic FNA results. FS was diagnostic in 104/130 (80%) patients and non-diagnostic in 26/130 (20%) patients. Sensitivity, specificity, positive and negative predictive values and positive and negative likelihood ratios were 81.3%, 97.8%, 96%, 88%, 36.95 and 0.19 respectively for FNA; 100%, 61.3%, 61.7%, 100%, 2.58 and 0 respectively for MIBI; and 80.5%, 100%, 100%, 89%, 0 and 0.2 respectively for FS. Use of both MIBI scans and FS in patients with non diagnostic FNA rendered a specificity and sensitivity of 100%. CONCLUSION: MIBI scans exclude malignancy in a significant proportion of patients with non diagnostic FNAs (38% in this study). Cystic nodules with a positive MIBI scan should be further investigated even when the FNA result indicates a benign lesion. Combined use of FNA, MIBI and FS offers the best diagnostic accuracy. PMID- 15133638 TI - MRI findings of myositis in Behcet disease. AB - We report the magnetic resonance imaging findings in a case of localized myositis in a 23-year-old man with long-standing Behcet disease. PMID- 15133639 TI - Chondroid lipoma of the trunk: MRI appearance and pathologic correlation. AB - Chondroid lipoma is a rare tumour of adipose tissue, bearing a strikingly close pathologic resemblance to myxoid liposarcoma and extraskeletal myxoid chondrosarcoma. Unlike these malignant tumours, chondroid lipoma has a non aggressive behaviour and does not require radical treatment. Although repeatedly reported in the proximal extremities and limb girdles, this rare entity may less frequently be observed in the trunk. We describe the imaging findings of a chondroid lipoma in the trunk and provide a discussion on the radiologic pathologic correlation and differential diagnosis. PMID- 15133640 TI - Sonography of injury of the ulnar collateral ligament of the elbow-initial experience. AB - OBJECTIVE: The purpose of this study is to describe the sonographic appearance of injuries of the ulnar collateral ligament (UCL) of the elbow. DESIGN AND PATIENTS: Eight non-professional male baseball pitchers, ages 13-35 years, with medial elbow pain and clinical suspicion of ulnar collateral ligament injury, were referred for imaging. All eight underwent sonography of the affected and contralateral asymptomatic elbow, and six also underwent MR imaging. Neither valgus stress nor power Doppler was used during the sonographic examinations. Time from onset of symptoms to imaging was 1.5 weeks to 6 months. Three patients had surgical confirmation of their injuries, with time from imaging to surgery of 2 days to 9 months. RESULTS: In four patients, the UCL was ruptured, manifest sonographically in three cases as discontinuity of the normally hyperechoic ligament with anechoic fluid in the gap and in one case as non-visualization of the ligament with heterogeneous echogenicity in the expected location of the ligament. Two adolescent patients had avulsions of the UCL from the medial epicondyle, with sonographic demonstration of the avulsed echogenic bony fragment in both cases. One patient had a mild sprain, manifest as mild thickening and decreased echogenicity of the ligament sonographically compared with the contralateral normal elbow, with mild surrounding hypoechoic edema. The eighth patient had a small partial tear of the deep surface of the distal aspect of the ligament, visualized as a hypoechoic focus between the deep surface of the ligament and its ulnar attachment. CONCLUSION: Tears of the ulnar collateral ligament are manifested sonographically as non-visualization of the ligament or alteration of the normal morphology. PMID- 15133641 TI - Relation between bacterial strain resistance to solvents and biodesulfurization activity in organic medium. AB - Microorganisms used in biodesulfurization of petroleum products have to withstand high concentrations of hydrocarbons. The capacities of seven desulfurizing strains of Rhodococcus to be active in the presence of solvents were evaluated. Octanol and toluene (log P=2.9) were selected as toxic solvents. The effect of the solvents was determined by measuring either inhibition of growth or the decrease in respiratory activity of the cells. Differences among strains in their resistance to solvent responses were observed, but these variations were dependent on the test used. Resistance to solvents was then compared to the capacity of the different strains to retain biodesulfurization activity in the presence of hexadecane. Inhibition of desulfurization by high concentrations of hexadecane was found to be well correlated to the sensitivity of the strains to respiration inhibition by toluene, but not to growth inhibition. This result also showed that the respirometric test was a rapid and reliable test to select solvent-resistant strains for use as resting cells in biocatalysis processes, such as biodesulfurization, in organic media. PMID- 15133642 TI - Isolation and characterization of novel bacteria degrading polycyclic aromatic hydrocarbons from polluted Greek soils. AB - Three bacterial strains, designated as Wphe1, Sphe1, and Ophe1, were isolated from Greek soils contaminated with polycyclic aromatic hydrocarbon (PAH) containing waste from the wood processing, steel, and oil refinery industries. Wphe1, Sphe1, and Ophe1 were characterized and identified as species of Pseudomonas, Microbacterium, and Paracoccus, respectively, based on Gram staining, biochemical tests, phospholipid analysis, FAME analysis, G+C content and 16S rRNA gene sequence analysis. The results of gas chromatography showed that strain Wphe1 degraded naphthalene, phenanthrene, and m-cresol over a wide temperature range; strain Sphe1 was a degrader of phenanthrene and n-alkanes; most interestingly, strain Ophe1 degraded anthracene, phenanthrene, fluorene, fluoranthene, chrysene, and pyrene, as well as cresol compounds and n-alkanes as sole carbon source. This is the first report of a representative of the genus Paracoccus capable of degrading PAHs with such versatility. These three strains may be useful for bioremediation applications. PMID- 15133643 TI - Metabolite production during transformation of 2,4,6-trinitrotoluene (TNT) by a mixed culture acclimated and maintained on crude oil-containing media. AB - Metabolites formed during 2,4,6-trinitrotoluene (TNT) removal by a mixed bacterial culture (acclimated and maintained on crude oil-containing medium and capable of high rates of TNT removal) were characterized. In resting cell experiments in the absence of glucose, 46.2 mg/l TNT were removed in 171 h (87.5% removal), with a combined total formation of 7.7 mg/l amino-4,6-dinitrotoluene (ADNT) and 0.3 mg/l 4,4'-azoxytetranitrotoluene and 2',4-azoxytetranitrotoluene, leaving 70% of the initial TNT unaccounted for. In the presence of glucose, resting cells removed 45.4 mg/l TNT in 49 h (95.5% removal), with 9.1 mg/l ADNT and 2.4 mg/l azoxy compounds being produced, leaving 70.3% of the TNT unaccounted for. Growing cells (glucose present) were capable of removing 44.2 mg/l TNT within 21 h (97.9% removal), with the concomitant formation of 1.8 mg/l ADNTs and 2.2 mg/l azoxy compounds. Denitrated TNT in the form of 2,6-dinitrotoluene was also produced in growing cells with a maximum amount of 1.31 mg/l after 28 h, followed by a slight decrease with time, leaving 88.5% of the initial TNT unaccounted for after 171 h. Radiolabeled (14)C-TNT studies revealed 4.14% mineralization after an incubation period of 163 days with growing cells. PMID- 15133644 TI - Chemical and toxic evaluation of a biological treatment for olive-oil mill wastewater using commercial microbial formulations. AB - Olive-oil-mill wastewater (OMW) has significant polluting properties due to its high levels of chemical oxygen demand (COD), biochemical oxygen demand (BOD), and phenols. In the present study, different commercial bacterial formulations were used in the biological treatment of OMW. COD and toxicity testing using primary consumers of the aquatic food chain (the rotifer Brachionus calyciflorus and the crustacean Daphnia magna) were employed to evaluate abatement of the organic load and reduction of the toxic potential. In addition, the four most active formulations were tested mixed pair-wise on the basis of their unique characteristics in order to evaluate the improvement of treatment. The effect of treatment was assessed by measuring COD removal, reduction of total phenols, and decreased toxicity. The results obtained with the mixed formulations showed that the maximum removal of the organic load was about 85%, whereas phenols were reduced by about 67%. The toxicity for rotifers decreased by 43% and for crustaceans by about 83%. PMID- 15133645 TI - HLA-G and IL-10 in serum in relation to HLA-G genotype and polymorphisms. AB - The expression and importance of the non-classical human leukocyte antigen (HLA) class Ib gene, HLA-G, at the feto-maternal interface have been recognized. The HLA-G molecule is almost monomorphic and expressed in both membrane-bound and soluble isoforms. It has been shown to inhibit NK-mediated cell lysis and influence cytokine expression. Recently, a possible boarder immunoregulatory function of HLA-G also in adult life has been recognized. HLA-G gene polymorphism has been linked to differences in gene expression profile of alternatively spliced HLA-G transcripts and levels of specific HLA-G mRNA isoforms. On this background it is of general interest to further elucidate any associations between HLA-G polymorphism and protein expression. We have HLA-G genotyped 85 individuals attending IVF treatment, and further studied sHLA-G1/HLA-G5 and interleukin-10 (IL-10) in serum samples. In 21% of the serum samples sHLA-G1/HLA G5 could be detected. There was no correlation between sHLA-G1/HLA-G5 and IL-10 concentrations in serum. Soluble HLA-G1/HLA-G5 was not detected in any samples homozygous for a 14-bp insertion polymorphism in exon 8 of the 3'-untranslated region (3'UTR) of the HLA-G gene ( P=0.03; Fisher's exact test). Polymorphisms in the 5'-upstream regulatory region (5'URR) of the HLA-G gene were also studied. In conclusion, this study indicates that polymorphisms in the 3'UTR and the 5'URR of the HLA-G gene may influence the expression of sHLA-G of possible importance in pathological pregnancies and also in organ transplantation. PMID- 15133646 TI - Isolation of monotreme T-cell receptor alpha and beta chains. AB - Monotremes are an ancient mammalian lineage that last shared a common ancestor with the marsupial and eutherian (placental) mammals about 170 million years ago. Characterization of their immune genes is allowing us to gain insights into the evolutionary processes that lead to the 'mammalian' immune response. Here we describe the characterization of the first cDNA clones encoding T-cell receptors from a monotreme. Two TCR alpha-chain cDNAs ( TCRA) from the short-beaked echidna, Tachyglossus aculeatus, containing complete variable, joining and constant regions were isolated. The echidna TCRA constant region shares approximately 37% amino acid identity with other mammalian TCRA constant region sequences. The two variable regions belong to the TCRAV group C, which also contains V genes from humans, mice, cattle and chickens. One echidna TCR beta chain cDNA ( TCRB) containing the entire constant region was isolated and sequenced. It shares about 63% identity with other mammalian TCRB constant region sequences. The echidna TCRBV belongs to TCRBV group A, which also contains V genes from various eutherian species. Southern blot analysis indicates that, like in other mammalian species, there is only one TCRA constant region copy in the echidna genome, but at least two TCRB constant regions. PMID- 15133647 TI - Significance of low-frequency local fluctuation motions in the transmembrane B and C alpha-helices of bacteriorhodopsin, to facilitate efficient proton uptake from the cytoplasmic surface, as revealed by site-directed solid-state 13C NMR. AB - 13C NMR spectra of [1-13C]Val- or -Pro-labeled bacteriorhodopsin (bR) and its single or double mutants, including D85N, were recorded at various pH values to reveal conformation and dynamics changes in the transmembrane alpha-helices, in relation to proton release and uptake between bR and the M-like state caused by modified charged states at Asp85 and the Schiff base (SB). It was found that the D85N mutant acquired local fluctuation motion with a frequency of 10(4) Hz in the transmembrane B alpha-helix, concomitant with deprotonation of SB in the M-like state at pH 10, as manifested from a suppressed 13C NMR signal of the [1-13C] labeled Val49 residue. Nevertheless, local dynamics at Pro50 neighboring with Val49 turned out to be unchanged, irrespective of the charged state of SB as viewed from the 13C NMR of [1-13C]-labeled Pro50. This means that the transmembrane B alpha-helix is able to acquire the fluctuation motion with a frequency of 10(4) Hz beyond the kink at Pro50 in the cytoplasmic side. Concomitantly, fluctuation motion at the C helix with frequency in the order of 10(4) Hz was found to be prominent, due to deprotonation of SB at pH 10, as viewed from the 13C NMR signal of Pro91. Accordingly, we have proposed here a novel mechanism as to proton uptake and transport based on a dynamic aspect that a transient environmental change from a hydrophobic to hydrophilic nature at Asp96 and SB is responsible for the reduced p Ka value which makes proton uptake efficient, as a result of acquisition of the fluctuation motion at the cytoplasmic side of the transmembrane B and C alpha-helices in the M-like state. Further, it is demonstrated that the presence of a van der Waals contact of Val49 with Lys216 at the SB is essential to trigger this sort of dynamic change, as revealed from the 13C NMR data of the D85N/V49A mutant. PMID- 15133648 TI - Speckle interferometry applied to pharmacodynamic studies: evaluation of parasite motility. AB - The work reported here describes the application of the optical technique known as dynamic speckle interferometry to evaluate the motility of nematode parasites exposed to different anthelmintic drugs. This technique, a well proven tool for assessing the time evolution of different phenomena, is here successfully used to quantify parasite motility in pharmacodynamic assays. The characterization of the pharmacological properties of anthelmintic drugs is critical to optimize their use in parasite control. Besides, the evaluation of nematode motility is a relevant indicator of the pharmacodynamic effect of anthelmintic drugs. The application of this approach to study the motility of Haemonchus contortus (used as a model of nematode parasites) larvae exposed to different drugs is presented, showing its usefulness. PMID- 15133650 TI - Comments on "MRI of the tectorial and posterior atlanto-occipital membranes in the late stage of whiplash injury" ( Neuroradiology, 2003, 45:585-591). PMID- 15133652 TI - Characterization of the adsorption of omega-(thiophene-3-yl alkyl) phosphonic acid on metal oxides with AR-XPS. AB - The aim of the work discussed in this paper was to characterize adsorbed self assembled monolayers on different metal oxide substrates with angle-resolved XPS measurements. The substrates used were silicon wafers (100) coated with 300 nm Al, Ta, or Ti. They were coated with acids by immersing them in an ethanol solution. The orientation of long-chain organic acids adsorbed on metal oxides has been successfully identified by angle-resolved XPS. On Al, Ta, and Ti substrates, C(11) chains are orientated in the right manner, i.e. with the phosphonic group at the bottom and the thiophene group on top. The orientations of the C(2) and C(6) chains are not clear. The thickness of the layers could be obtained by using Tougaard nanostructure analysis, and it shows monolayers. A model of the chemical bonds between the phosphonic group and the metal could be developed from the chemical shift. For titanium, all three P-O bonds bind to the metal substrate, whereas only the P-O(H) bond binds to the metal on aluminium and tantalum. PMID- 15133653 TI - Identification of steel by X-ray fluorescence analysis with a pyroelectric X-ray generator. AB - An application of X-ray fluorescence analysis with a pyroelectric X-ray generator is presented. Steel standard samples were identified by X-ray fluorescence analysis with this novel X-ray generator to check its capability for performing qualitative and quantitative analysis as an X-ray source for X-ray fluorescence spectrometers. Cr, Ni, V, Co, and W were detected in steel standard samples. V and Cr can be detected even when the content is below 1%. Although it is difficult to detect minor elements because of the low power of the excitation X rays, it is possible to identify the analyzed samples on the basis of major elements at the percentage level. The pyroelectric X-ray generator is very suitable for portable X-ray fluorescence spectrometers. PMID- 15133651 TI - Preserved prism adaptation in bilateral optic ataxia: strategic versus adaptive reaction to prisms. AB - To date the anatomical substrate(s) of prism adaptation remain(s) particularly debated, with two main candidates emerging from the literature: the posterior parietal cortex (PPC) and the cerebellum. The functional processes involved in the acquisition of the adaptive aftereffects also remain largely unknown. The main result shown here is that a patient with a bilateral optic ataxia can adapt to an optical deviation, which allows us to make a step forward on these two issues. First, it demonstrates that the corresponding part of the PPC is not a necessary substrate for prism adaptation. Second, since this patient exhibits deficit for fast visuo-motor guidance, it provides direct evidence for a dissociation between on-line visuo-motor control and visuo-motor plasticity. Since the intermanual transfer rate of adaptation is larger in this patient than in control subjects, the PPC may still have an influence on adaptation under normal conditions. We propose a model of the relative contribution of the PPC and the cerebellum during prism exposure, associating these two structures with the two interacting behavioural components of prism adaptation described by previous psychophysical experiments: the strategic component would be linked to the PPC and the adaptive component to the cerebellum. In this model, the strategic component enters in conflict with the development and the generalisation of the adaptive aftereffects. This idea is compatible with the fact that a lesion of the PPC increases the transfer rate and the generalisation of the adaptation, as is also observed in unilateral neglect. PMID- 15133654 TI - Segmental duplication associated with the human-specific inversion of chromosome 18: a further example of the impact of segmental duplications on karyotype and genome evolution in primates. AB - The human-specific pericentric inversion of chromosome 18 was analysed using breakpoint-spanning BACs from the chimpanzee and human genome. Sequence and FISH analyses disclosed that the breakpoints map to an inverted segmental duplication of 19-kb, which most likely mediated the inversion by intrachromosomal homologous recombination. The 19-kb duplication encompasses the 3' end of the ROCK1 gene and occurred in the human lineage. Only one copy of this segment is found in the chimpanzee. Due to the inversion, the genomic context of the ROCK1 and USP14 genes is altered. ROCK1 flanks USP14 in the long arm of the chimpanzee chromosome 17, which is homologous to human chromosome 18. This order is interrupted by the inversion in humans. ROCK1 is localized close to the pericentromeric region in 18q11 and USP14 is inverted to distal 18p11.3 in direct neighbourhood to LSAU satellites, beta-satellites and telomere-associated repeats. Our findings essentially confirm the analysis of Dennehey et al. (2004). Intriguingly, USP14 is differentially expressed in human and chimpanzee cortex as well as fibroblast cell lines determined previously by the analysis of oligonucleotide arrays. Either position effects mediated by the proximity to the telomeric region or nucleotide divergence in regulatory regions might account for the differential expression of USP14. The assignment of the breakpoint region to a segmental duplication underlines the significance of the genomic architecture in the context of genome and karyotype evolution in hominoids. PMID- 15133655 TI - Genetic interactions between an RNA polymerase II phosphatase and centromeric elements in Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiase protein phosphatase Fcp1 has been implicated in the regulation of transcription by RNA polymerase II, and is encoded by the essential gene FCP1. A screen was carried out for multicopy suppressors of the temperature sensitive phenotype of two phosphatase mutants, fcp1-2 and fcp1-4. Only the wild type FCP1 was found to suppress (complement) the fcp1-4 mutation. For fcp1-2 three second-site suppressors were identified. One contained the ORF for ZDS1. The remaining two suppressors mapped to the centromere regions of chromosomes I and V. Suppression due to centromere DNA was found to be more dependent on the CDEIII region than on other regions of the centromere. The presence of a suppressor centromere affected the level of Fcp1 protein and the overall phosphorylation state of RNA polymerase II (RNAPII) in fcp1-2 cells, but not wild type cells, grown at both permissive and non-permissive temperatures. In addition, genetic interactions were identified between this FCP1 mutant and the genes SKP1, CEP3 and CBF1, which code for centromere binding proteins. The mechanism of suppression and regulation of Fcp1-2 protein activity by centromeric DNA is discussed. PMID- 15133656 TI - Reverse genetic analysis of the glutathione metabolic pathway suggests a novel role of PHGPX and URE2 genes in aluminum resistance in Saccharomyces cerevisiae. AB - We have taken a systematic genetic approach to study the potential role of glutathione metabolism in aluminum (Al) toxicity and resistance, using disruption mutants available in Saccharomyces cerevisiae. Yeast disruption mutants defective in phospholipid hydroperoxide glutathione peroxidases (PHGPX; phgpx1 Delta, phgpx2 Delta, and phgpx3Delta), were tested for their sensitivity to Al. The triple mutant, phgpx1 Delta/2Delta/3Delta, was more sensitive to Al (55% reduction in growth at 300 microM Al) than any single phgpx mutant, indicating that the PHGPX genes may collectively contribute to Al resistance. The hypersensitivity of phgpx3Delta to Al was overcome by complementation with PHGPX3, and all PHGPX genes showed increased expression in response to Al in the wild-type strain (YPH250), with maximum induction of approximately 2.5-fold for PHGPX3. Both phgpx3Delta and phgpx1Delta/2Delta/3Delta mutants were sensitive to oxidative stress (exposure to H(2)O(2) or diamide). Lipid peroxidation was also increased in the phgpx1Delta/2Delta/3Delta mutant compared to the parental strain. Disruption mutants defective in genes for glutathione S-transferases (GSTs) (gtt1Delta and gtt2Delta), glutathione biosynthesis (gsh1Delta and gsh2Delta), glutathione reductase (glr1Delta) and a glutathione transporter (opt1Delta) did not show hypersensitivity to Al relative to the parental strain BY4741. Interestingly, a strain deleted for URE2, a gene which encodes a prion precursor with homology to GSTs, also showed hypersensitivity to Al. The hypersensitivity of the ure2Delta mutant could be overcome by complementation with URE2. Expression of URE2 in the parental strain increased approximately 2 fold in response to exposure to 100 microM Al. Intracellular oxidation levels in the ure2Delta mutant showed a 2-fold (non-stressed) and 3-fold (when exposed-to 2 mM H(2)O(2)) increase compared to BY4741; however, the ure2Delta mutant showed no change in lipid peroxidation compared to the control. The phgpx1Delta/2Delta/3Delta and ure2Delta mutants both showed increased accumulation of Al. These findings suggest the involvement of PHGPX genes and a novel role of URE2 in Al toxicity/resistance in S. cerevisiae. PMID- 15133658 TI - Changes in the level of antioxidants in the blood from mice infected with Trichinella spiralis. AB - The biological reaction caused by oxygen-derived free radicals at the molecular and cellular levels involves many different biochemical components which can be directly damaged by oxidizing radicals. As such a reaction may lead to pathological processes, defence mechanisms have evolved to limit the rate of free radical production. These mechanisms employ low-molecular-weight non-enzymatic antioxidants and antioxidant enzymes which are inducible by oxidant stress. In this study, the activity of two antioxidant enzymes, superoxide dismutase (EC 1.15.1.1) and glutathione peroxidase (EC 1.11.1.9), and the level of non enzymatic antioxidants (total antioxidant status) in the blood from mice infected with Trichinella spiralis was examined. We observed a statistically significant, up to above twofold increase (relative to the control value in uninfected mice) in the level of both enzymes as well as in the total antioxidant status. An intensification of antioxidant processes during trichinellosis could be related to the presence of T. spiralis larvae, which may induce phagocytes to generate free radicals. Our research shows that the maximum growth in antioxidant activity in the blood appears during the period of the greatest muscle damage caused by T. spiralis infection at 3-7 weeks post-infection. PMID- 15133659 TI - Parasitic helminth fauna of the cutlass fish, Trichiurus lepturus L., and the differentiation of four anisakid nematode third-stage larvae by nuclear ribosomal DNA sequences. AB - The helminth fauna of the gastrointestinal tract and abdominal cavity of cutlass fish, Trichiurus lepturus L., off the Taiwanese coast of the north-western Pacific was investigated. The following helminths were found: (1) nematodes- Anisakis simplex, Hysterothylacium aduncum, Porrocaecum decipiens, Raphidascaris trichiuri; (2) digeneans--adult Lecithochirium trichiuri; and (3) cestodes plerocercoids of Proteocephalus spp. The third-stage larvae of these four anisakid nematodes were characterized genetically using a molecular approach. The nuclear ribosomal DNA region spanning the first internal transcribed spacer (ITS 1), the 5.8S gene and the second internal transcribed spacer (ITS-2) was amplified and sequenced. Based on the sequence differences, a PCR-based restriction fragment length polymorphism method was established for the unequivocal delineation of the four species. Phylogenetic analysis showed that H. aduncum clustered with P. decipiens, whereas A. simplex was not closely related to these according to the nucleotide sequences of all rDNA. PMID- 15133660 TI - The musculature and associated innervation of adult and intramolluscan stages of Echinostoma caproni (Trematoda) visualised by confocal microscopy. AB - Gross anatomy of muscle and sensory/motor innervation of adult and intramolluscan developmental stages of Echinostoma caproni have been investigated to ascertain the organisation and the functional correlates of any stage-specific patterns of staining. Using indirect immunocytochemistry to demonstrate neuroactive substances and the phalloidin-fluorescence technique for staining myofibril F actin, the muscle systems and aminergic and peptidergic innervation of daughter rediae, cercariae, metacercariae, and pre- and post-ovigerous adults were examined and compared using confocal scanning laser microscopy. A complex arrangement of specific muscle fibre systems occurs within the body wall (composed of circular, longitudinal and diagonal fibres), suckers (radial, equatorial, meridional), pharynx (radial, circular), gut caeca (mainly circular), cercarial tail (circular, pseudo-striated longitudinal), and ducts of the reproductive system (circular, longitudinal), presumed to serve locomotor, adhesive, alimentary and reproductive functions. Immunostaining for serotonin (5 HT) and FMRFamide-related peptides (FaRPs) was evident throughout the central (CNS) and peripheral (PNS) nervous systems of all stages, and use of dual labelling techniques demonstrated separate neuronal pathways for 5-HT and FaRP in both CNS and PNS. FaRP expression in the innervation of the ootype wall was demonstrated only in post-ovigerous worms and not in pre-ovigerous worms, suggesting an involvement of FaRP neuropeptides in the process of egg assembly. Comparison of the present findings with those recorded for other digeneans suggests that muscle organisation and innervation patterns in trematodes are highly conserved. PMID- 15133661 TI - Establishment of green fluorescent protein-expressing hepatocellular carcinoma cell lines with different metastatic potential: relevant models for in vivo monitoring of metastasis and angiogenesis. AB - PURPOSE: To establish stable green fluorescent protein (GFP)-expressing metastatic human hepatocellular carcinoma (HCC) cell lines with different metastatic potential for long-term in vivo studies of metastasis and angiogenesis. METHODS: The pIRES2-EGFP vector, which contains an enhanced GFP gene, was transfected into MHCC97-H and MHCC97-L, HCC cell lines with different metastatic potential. The stability of GFP expression, basic biological characteristics, invasion abilities in vitro, and spontaneous metastasis in vivo of the new cell lines (MHCC97-HG and MHCC97-LG) were studied. Microvessel density (MVD) of orthotopic implanted tumors was compared by anti-CD31 immunohistochemical staining, and real-time angiogenesis and metastasis of GFP transfected tumors were detected by intravital fluorescent microscope. RESULTS: The GFP-transfected cell lines stably expressed green fluorescence in the absence of G418 over a 36-day period. Compared with the parental cell lines, they exhibited no distinct differences in biological characteristics. MHCC97-HG showed more aggressive invasion and spontaneous metastatic behavior than MHCC97-LG, and even its parental cell line, MHCC97-H (P<0.01). MVD levels induced by MHCC97-HG orthotopic implanted tumors were significantly higher than MHCC97-LG (P<0.01). Real-time angiogenesis and sequential steps of metastasis could be detected clearly under intravital fluorescent microscope. CONCLUSIONS: These two stable GFP-expressing HCC cell lines with the same genetic background and different metastatic potential were established, which could be useful models for monitoring metastasis and angiogenesis of HCC. PMID- 15133662 TI - The association of the expression level of protein tyrosine phosphatase PRL-3 protein with liver metastasis and prognosis of patients with colorectal cancer. AB - PURPOSE: To investigate PRL-3 protein expression in normal colorectal epithelia and colorectal cancers with monoclonal antibody (MAb) against PRL-3. METHODS: MAb against PRL-3 was prepared with the hybridoma technique, and its specificity was confirmed with ELISA and Western blotting assays. The expression of PRL-3 protein in normal colorectal epithelia and colorectal cancers was examined by immunohistochemistry assay. Logistic regression and survival analysis were performed to determine the clinical significance of PRL-3 expression. RESULTS: MAb 3B6 against PRL-3 was obtained and showed high specificity. PRL-3 protein was expressed in two of 28 (7.1%) normal colorectal epithelia, 21 of 88 (23.9%) primary colorectal cancers, 22 of 41 (53.7%) metastatic lymph nodes and eight of 12 (66.7%) liver metastases, respectively. The PRL-3 expression rates of metastases were significantly higher than those of primary colorectal cancers and normal colorectal epithelia (P < 0.05). PRL-3 expression was significantly associated with the liver metastasis of colorectal cancer (P = 0.004) and tended to shorten survival time (P = 0.0145). CONCLUSIONS: This is the first study demonstrating that PRL-3 is a potential marker for liver metastasis of colorectal cancer and negatively influences the prognosis of colorectal cancer patients. PMID- 15133663 TI - Oxidative DNA damage in placentas from normal and pre-eclamptic pregnancies. AB - Placental oxidative stress was suggested to play a role in the pathogenesis of pre-eclampsia (PE). In this study, levels of 8-hydroxy-2'-deoxyguanosine (8-OH dG), a well-established marker of oxidative DNA damage, were analysed in placental cellular DNA from normal (group NP) and pre-eclamptic (group PE) pregnancies as well as from PE pregnancies complicated by intrauterine growth restriction (group PE-IUGR). Placental samples obtained immediately after delivery were frozen at -80 degrees C until analysis. Cellular DNA was isolated, hydrolysed and analysed using high-performance liquid chromatography. Native nucleosides were monitored at 254 nm and 8-OH-dG using electrochemical detection. Concentrations of 8-OH-dG were expressed as micro mol/mol 2'-deoxyguanosine. In group NP, mean concentration of 8-OH-dG reached 179.97+/-80.58 (+/-SEM; micro mol/mol dG). 8-OH-dG levels were higher in group PE (273.44+/-110.14 micro mol/mol), but the difference was not significant in comparison with group NP. Highest concentrations of 8-OH-dG were found in group PE-IUGR (428.97+/-141.40 micro mol/mol), with levels significantly higher than in group NP, but not group PE. The results indicate a positive correlation between the severity of PE and the degree of oxidative stress and corroborate previous studies suggesting reactive oxygen species to be involved in the pathophysiology of PE. PMID- 15133664 TI - Organization of Iroquois genes in fish. AB - In mammals, a total of six iroquois ( Irx) genes exist, which are organized into two clusters. Here we report on the organization of all iroquois genes present in fish, using zebrafish ( Danio rerio) and pufferfish ( Fugu rubripes and Tetraodon nigroviridis) as examples. A total of 10 Irx genes were found in pufferfish, and 11 in zebrafish; all but one of these genes are organized into clusters (four clusters plus one isolated gene locus). The "extra" fish clusters result from chromosome duplication in the fish lineage, after its divergence from tetrapod vertebrates. Two of the four fish clusters are highly conserved to the ones in mammals, with regard to similarity of genes and cluster architecture. Irx genes within the other two clusters have diverged in sequence and cluster organization, suggesting functional divergence. These results will allow us to use the zebrafish system for functional and comparative studies of iroquois genes in vertebrate development. PMID- 15133665 TI - In vivo visualization of Tradescantia leaf tissue and monitoring the physiological and morphological states under different water supply conditions using optical coherence tomography. AB - The optical coherence tomography (OCT) capabilities of plants were evaluated using leaves of Tradescantia pallida (Rose) D. Hunt. The internal structure of the leaf tissues was visualized in vivo and the physiological and morphological states of the tissues under different water supply conditions were monitored using OCT. The OCT technique provides non-invasive two-dimensional images directly on intact plants. The acquisition time of a two-dimensional image with a size of 200x200 pixels and a spatial resolution of 15 microm is 1-3 s. It was shown that OCT is a useful tool for monitoring the physiological and morphological states of plant tissues supplied with varying amounts of water and under the influence of different chemical factors. PMID- 15133666 TI - Dormancy of Arabidopsis seeds and barley grains can be broken by nitric oxide. AB - Seeds of Arabidopsis thaliana (L.) Heynh. and grains of barley ( Hordeum vulgare L.) were used to characterize the affects of nitric oxide (NO) on seed dormancy. Seeds of the C24 and Col-1 ecotypes of Arabidopsis are almost completely dormant when freshly harvested, but dormancy was broken by stratification for 3 days at 4 degrees C or by imbibition of seeds with the NO donor sodium nitroprusside (SNP). This effect of SNP on dormancy of Arabidopsis seeds was concentration dependent. SNP concentrations as low as 25 microM reduced dormancy and stimulated germination, but SNP at 250 microM or more impaired seedling development, including root growth, and inhibited germination. Dormancy was also reduced when Arabidopsis seeds were exposed to gases that are generated by solutions of SNP. Nitrate and nitrite, two other oxides of nitrogen, reduced the dormancy of Arabidopsis seeds, but much higher concentrations of these were required compared to SNP. Furthermore, the kinetics of germination were slower for seeds imbibed with either nitrate or nitrite than for seeds imbibed with SNP. Although seeds imbibed with SNP had reduced dormancy, seeds imbibed with SNP and abscisic acid (ABA) remained strongly dormant. This may indicate that the effects of ABA action on germination are downstream of NO action. The NO scavenger 2-(4-carboxyphenyl) 4,4,5,5-tetramethylimidazoline-1-oxyl-3 oxide (cPTIO) strengthened dormancy of unstratified and briefly stratified Arabidopsis seeds. Dormancy of three cultivars of barley was also reduced by SNP. Furthermore, dormancy in barley grain was strengthened by imbibition of grain with cPTIO. The data presented here support the conclusion that NO is a potent dormancy breaking agent for seeds and grains. Experiments with the NO scavenger suggest that NO is an endogenous regulator of seed dormancy. PMID- 15133667 TI - Palmitic acid metabolism in the soleus muscle in vitro in hypo- and hyperthyroid rats. AB - The aim of this study was to establish whether the rate of fatty acid (FA) incorporation and its utilization by the isolated soleus muscle is modified under conditions of thyroid hormone deficit or excess. The rate of palmitic acid (PA) uptake, oxidation and incorporation into intramuscular lipids with increasing PA concentration (0.5-1.5 mM) in the incubation medium were determined. In hypothyroid rats intramuscular triacylglycerol (TG) synthesis was increased, while the rate of PA oxidation to CO2 and incorporation into mono- and diacylglycerols (MG/DG) and phospholipids (PL) remained unchanged. In rats with triiodothyronine (T3) excess the rate of all processes studied was enhanced, although the percentage incorporation of PA into different classes of intramuscular lipids was fairly constant and, independently of thyroid state and FA concentration in the medium, was 56-66% for TG, 9-14% for MG/DG and 24-32% for PL. Our results thus indicate that even short-term T3 excess accelerates the rate of FA uptake and metabolism in the oxidative soleus muscle, whereas in hypothyroid rats only intramuscular TG synthesis is affected. PMID- 15133668 TI - Regulation of GABA release by depolarisation-evoked Ca2+ transients at a single hippocampal terminal. AB - We correlated dynamic changes in free cytosolic [Ca2+] ([Ca2+]i) within single presynaptic terminals of cultured hippocampal neurones with the postsynaptic GABA mediated currents. The local changes in [Ca2+]i and evoked inhibitory postsynaptic currents (eIPSCs) were recorded simultaneously using Fura-2 fluorescence and whole-cell patch-clamp respectively. The Ca2+ signals and eIPSCs were evoked by direct extracellular electrical stimulation of a single presynaptic terminal by short depolarising pulses. The presynaptic Ca2+ transient was graded by varying the amplitude of extracellular stimulating pulses. The probability of the release event, P, estimated for each stimulation strength, reached a maximum (P=1) when the Ca2+ signal became maximal and remained at this level at higher stimulation strength, despite the subsequent decrease in the amplitude of the Ca2+ transient. A gradual, linear increase in stimulation amplitude (Vstim) resulted in a bell-shaped dependence of the averaged amplitudes of Ca2+ signals and corresponding averaged amplitudes of eIPSCs. Analysis of the eIPSC demonstrated that the decrease in both the mean eIPSC amplitude and the mean quantal content of release resulted from a reduction in the probability of multivesicular release, i.e. in the disappearance of failures and in the decrease of individual eIPSC amplitude. The Ca2+ signals of similar amplitude resulted in both random and determinate (non-random) neurotransmitter release. We conclude that depolarisation-induced elevation of [Ca2+]i within the terminal is necessary but not sufficient for activation of vesicular release of neurotransmitter. PMID- 15133670 TI - Hybrid fibres under slow-to-fast transformations: expression is of myosin heavy and light chains in rat soleus muscle. AB - The aim of this study was to examine the expression pattern of myosin heavy chain (MHC) and myosin light chain (MLC) isoforms in single fibres from the rat soleus muscle under control (Cont) conditions and under conditions inducing slow-to-fast phenotype transitions. Two models of muscle phenotype modification, namely 2 weeks clenbuterol (CB) administration or hindlimb unloading (HU), were chosen to achieve a full range of appearance of hybrid fibres, i.show $132#e. fibres co expressing slow and fast myosin isoforms. MHC and MLC compositions were analysed in parallel by one-dimensional-gel electrophoresis. We showed that (i) the slow to-fast fibre type transitions at the MHC level were accompanied by exchanges of slow with fast MLC isoforms and (ii) that these transitions were characterized by increased proportions of hybrid profiles of both MHC and MLC isoforms, under both CB (27.5%) and HU (18%) conditions when compared with Cont (7%). This suggested a MHC-MLC coordinated program for myosin regulation during fibre type transitions. However, mismatched hybrid co-expression of MHC and MLC was also observed, probably resulting from differences in post-transcriptional regulation. Finally, in all the muscle fibre groups, specific favourable correlations between one MHC (IIa, IId or IIb) and one MLC (regulatory or essential) type were found. PMID- 15133669 TI - The 2P-domain K+ channels: role in apoptosis and tumorigenesis. AB - Two-pore (2P)-domain K+ channels have been shown recently to play a critical role in both cell apoptosis and tumorigenesis. The activity of two-pore, (TWIK) related acid-sensitive-3 (TASK-3) K+ channels, is responsible for K+-dependent apoptosis of cultured cerebellar granule neurons. Neuron death can be prevented by conditions that specifically reduce K+ efflux through the TASK-3 channels. Moreover, genetic transfer of TASK subunits into hippocampal neurons that lack TASK-3, induces apoptosis. These results indicate a direct link between TASK K+ channel activity and the physiological process of programmed cell death. The TASK 3 K+ channel gene has also been shown to be amplified genomically and over expressed in a significant number of breast tumours. TASK-3 has a potent oncogenic potential that appears to be related directly to its K+ channel function. In the present review, we will examine the pro-apoptotic and oncogenic properties of TASK-3. We will discuss: (1) the molecular and functional properties of the novel family of mammalian 2P domain K+ channels; (2) the role of TASK-3 in cerebellar granule neuron apoptosis and (3) the role of TASK-3 in breast tumorigenesis. PMID- 15133671 TI - Laparoscopic cholecystectomy: early and late complications and their treatment. AB - Laparoscopic cholecystectomy gained wide acceptance as treatment of choice for gallstone disease and cholecystitis. With this new technique, not only did the new era of minimal invasive surgery begin, but also the spectrum of complications changed. Laparoscopy-related complications such as access injuries and procedure related problems are discussed in our article. Typical mishaps are reviewed according to the literature. Set-up of the pneumoperitoneum (morbidity up to 0.2%); bleeding-from trocar sites and vascular injury (mortality up to 0.2%); biliary leaks and bile duct injuries are the main topics in this article (still on a level of 0.2%-0.8%). Aetiology, diagnosis and treatment are discussed, and an overview of the most cited classifications of bile duct injuries is summarised graphically. Finally, bowel injuries as a specific complication in laparoscopy are discussed (incidence up to 0.87%). CONCLUSION: Careful selection of patients, the knowledge of typical procedure-related complications, and their best treatment are the key points for a safe laparosopic cholecystectomy. PMID- 15133672 TI - Ulcerative colitis: conservative management and long-term effects. AB - Ulcerative colitis is characterized by chronic inflammation of the colon. Typical symptoms are diarrhoea, rectal bleeding, abdominal pain and fever. The aetiology of the disease is unclear. The inflammation can be localized in the rectum or can extend to the left side or the whole colon. Treatment for induction and remission maintenance depends on the severity and extension of mucosal inflammation. Topical 5-aminosalicylates have been shown in studies to be the treatment of choice in mild to moderate ulcerative colitis. Oral 5-aminosalicylates can be used in distal, mild and moderate ulcerative colitis and for remission maintenance. For patients with a more extended or severe inflammation, oral or i.v. corticosteroids should be used. Patients with severe and/or chronic disease require immunosuppressive therapy with azathioprine or 6-mercaptopurine. For patients with severe, chronic, refractory disease, cyclosporine i.v. can be used. If no response to treatment is seen, proctocolectomy should be considered. Biological agents such as beta-Interferon seem to be effective in mild to moderately ulcerative colitis, but further studies have to be performed. PMID- 15133673 TI - Small-incision (mini-laparotomy) versus laparoscopic cholecystectomy: a retrospective study in a university hospital. AB - BACKGROUND AND AIMS: Since the introduction of laparoscopic cholecystectomy into general practice in 1990, it has rapidly become the dominant procedure for gallbladder surgery. The aim of this study was to compare the results of the laparoscopic, open and mini-laparotomy approaches to cholecystectomy. PATIENTS AND METHODS: Our study covers a period of 6 years. A total of 1,276 patients underwent cholecystectomy for calculous biliary disease. The laparoscopic procedure was applied to 952 (74.6%) patients, while 210 (16.5%) underwent the traditional open cholecystectomy and the remaining 114 (8.9%) patients underwent mini-laparotomy cholecystectomy. RESULTS: Thirty-seven patients (3.9%) from the laparoscopic group required conversion to open cholecystectomy. Morbidity was similar in the open and laparoscopic groups (3.8%), while it was significantly lower in the mini-laparotomy group (0.8%). No major bile duct injuries occurred after the open or mini-laparotomy approaches. The median operation time was significantly shorter in the mini-laparotomy group than in the laparoscopic group (46 min vs 61 min). Hospital stay was significantly longer for the open cholecystectomy group (mean value 5.1 days) compared with the laparoscopic and mini-laparotomy groups (mean values 2.5 days and 2.7 days, respectively). Hospital expenses showed a saving of 786 Euro for each patient who underwent the open procedure and 980 Euro for each patient who underwent the mini-laparotomy approach compared with the laparoscopic one. CONCLUSION: We believe that commissioners of healthcare should question whether the benefits of laparoscopic cholecystectomy justify the additional cost after the introduction of the mini laparotomy approach. PMID- 15133674 TI - Survival after surgical treatment of early gastric cancer, surgical techniques, and long-term survival. AB - Early gastric cancer (EGC) is well accepted as having a favorable prognosis after surgical treatment. Difference in treatment strategies for EGC between Japan and western countries indicates a need for current information to be evaluated with regard to long-term survival rates of EGC patients throughout the world. To analyze survival rates and recurrence after resection of EGC, we investigated 51 reports in English that each included more than 50 cases of EGC treated by gastrectomy and had been published during the past 12 years (1992-2003). Prevalence of EGC among all gastric cancers was 45%-51% in Japan, but only 7%-28% in western countries. Mean age at diagnosis was less than 60 years in Japan and Korea, but was more than 60 in most of the Western countries. Actuarial and disease-specific 5-year survival rates for EGC were 72%-95.8% and 88%-98.3%, respectively. Those for EGC that were invading the submucosal layer were 71.6% 94.1% and 82%-96.6%, respectively. Those for EGC with lymph node metastasis were 57%-89.1% and 72%-93.5%, respectively. Prevalence of recurrence ranged from 1.0% to 13.8%. Larger clinical series with more EGC cases showed a lower prevalence of recurrence (P=0.531, P=0.0026). Liver and blood-borne distant metastasis represented the predominant pattern of relapse, accounting for over half (54%). Local recurrence and peritoneal dissemination represented 20% and 18% of all recurrences, respectively. Clinicopathological studies have shown lymph node metastasis to be closely related to depth of invasion, size of lesion, histological type, presence of ulcer or ulcer scar, and vessel involvement. Information on these factors is the key to successful treatment of EGC. When sufficient information has been assessed preoperatively, surgeons can select patients for whom less-invasive surgery should not increase the risk of recurrence. PMID- 15133675 TI - Modelling fire-fighter responses to exercise and asymmetric infrared radiation using a dynamic multi-mode model of human physiology and results from the sweating agile thermal manikin. AB - In this study, predicted dynamic physiological responses are compared with wear trials results for firefighter suits: impermeable (A), semi-permeable (B) and permeable (C), and underwear. Wear trials consisted of three rest phases and two moderate work phases, with a frontal infrared (IR) radiation exposure of 500 W/m2 for the last 15 min of each work phase. Simulations were performed by detailed modelling of the experimental boundary conditions, including the inhomogeneous IR radiation combined with clothing properties for still and walking conditions measured using the Sweating Agile thermal Manikin. Accounting for the effect of sweat gland activity suppression with increased skin wettedness, the predicted total moisture loss was insignificantly different (P<0.05) from the wear trial value for suits B and C but was 37% too high for suit A. Predicted evolution of core, mean skin and local skin temperatures agreed well with the wear trial results for all clothing. Root mean square deviations ranged from 0.11 degrees C to 0.26 degrees C for core temperatures and from 0.28 degrees C to 0.38 degrees C for mean skin temperatures, which where typically lower than the experimental error. Transient thermodynamic processes occurring within suit A may account for the delayed/reduced fall in core temperature following exercise. PMID- 15133676 TI - Selection of military survival gears using thermal manikin and computer survival model data. AB - This study presents a practical example of the selection of protective equipment for 12-h cold survival on land and at sea using computer model and manikin data. The thermal immersion manikin was exposed to 19 realistic survival scenarios to estimate the thermal resistance of different survival systems. The computer survival model used specific environmental limits and anthropometric data from the target population in addition to the estimated manikin thermal resistance values to generate survival times. The results showed that the required 12-h survival time criteria were met for all dry land scenarios (> 2 Clo), but not for wet land or water scenarios ( < 1 Clo). Those data provided the basis for the selection of survival equipment and the development of survival strategies for aircrew. PMID- 15133677 TI - Visual evoked potentials in workers with chronic solvent encephalopathy. AB - OBJECTIVES: Two promising variations of visual evoked potentials (VEPs) were studied in solvent-exposed workers: the effect of a low-contrast stimulus in comparison with the usually applied high contrast, and the ability of pattern onset VEP to reveal damage to specific visual cortical areas. In addition, we studied disturbances of a visual event-related potential (P300). METHODS: Thirty male patients (48 +/- 9 years of age) with solvent-induced chronic encephalopathy, and 41 controls (46 +/- 8 years) without solvent exposure, participated. Pattern-reversal checkerboards with low (11%) contrast and with high (93%) contrast between the checks were used. For onset VEPs two dedicated stimulus patterns were used. P300 was elicited with an "oddball" paradigm. RESULTS: At low contrast the N75-P100 peak-to-peak amplitude in the controls was 9.6 +/- 4.9 microV, i.e. 57% of the amplitude at high contrast (16.3 +/- 7.2 microV). In the patients the response at low contrast was only 48% of that at high contrast; the corresponding amplitudes were 7.5 +/- 3.5 microV and 15.8+/ 4.9 microV. For the pattern-onset VEPs no effect of exposure was found. With regard to the P300, the patients missed more targets (average 3.6%) than did the controls (average 0.5%). Patients had a smaller P300 amplitude (8.8 +/- 4.5 microV) than the controls (11.5 +/- 5.3 microV), and a longer latency (390 +/- 34 ms compared to 376 +/- 24 ms). CONCLUSION: The results point to a physiological basis for the solvent-induced decrease of visual contrast sensitivity as found by others by means of psychophysical methods. The results also suggest that the neurophysiological examination of the visual system in persons who have undergone exposure to toxins might be benefited by the addition of low-contrast stimuli. PMID- 15133678 TI - Intracellular localization of the cyclin-dependent kinase inhibitor p21CDKN1A-GFP fusion protein during cell cycle arrest. AB - The cyclin-dependent kinase (CDK) inhibitor p21CDKN1A is known to induce cell cycle arrest by inhibiting CDK activity and by interfering with DNA replication through binding to proliferating cell nuclear antigen. Although the molecular mechanisms have been elucidated, the temporal dynamics, as well as the intracellular sites of the activity of p21 bound to cyclin/CDK complexes during cell cycle arrest, have not been fully investigated. In this study we have induced the expression of p21CDKN1A fused to green fluorescent protein (GFP) in HeLa cells, in order to visualize the intracellular localization of the inhibitor during the cell cycle arrest. We show that p21-GFP is preferentially expressed in association with cyclin E in cells arrested in G1 phase, and with cyclin A more than with cyclin B1 in cells arrested in the G2/M compartment. In addition, we show for the first time that p21-GFP colocalizes with cyclin E in the nucleolus of HeLa cells during the G1 phase arrest. PMID- 15133679 TI - Purtscher-like retinopathy and Horner's syndrome following coil embolization of an intracavernous carotid artery aneurysm. AB - BACKGROUND: Coil embolization is a non-invasive method for treating intracranial vascular malformations by inducing thrombus formation. It is particularly useful in management of cerebral aneurysms and avoids the risks associated with surgical clipping. Occasionally, embolic complications occur which result in transient or permanent loss of vision. METHODS: Case report of a 29-year-old Caucasian woman who underwent coil embolization and balloon occlusion of an intracavernous carotid aneurysm. Shortly thereafter she described visual changes and a droopy eyelid. RESULTS: Examination revealed pupillary miosis and mild ptosis of the right upper lid. Perimetry showed an arcuate scotoma superiorly and an inferonasal step in the right eye. Fundus examination revealed multiple cotton wool spots along the peripapillary area and along the temporal vascular arcades reminiscent of Purtscher retinopathy. After 4 weeks, she had marked improvement in her visual symptoms. Most of the cotton-wool spots had resolved and the visual field had normalized. However, the ptosis and anisocoria remained unchanged. CONCLUSIONS: Coil embolization is a non-invasive method for treating carotid aneurysms. However, it carries the risk of ophthalmic events, warranting baseline ophthalmic examinations prior to such intervention. PMID- 15133680 TI - Sudden cardiac death in hereditary hemochromatosis: an underestimated cause of death? AB - Hereditary hemochromatosis (HH) is a frequent autosomal recessive disease which causes iron-overload of various organs. Of all northern European affected individuals, 90-95% show 1 of 3 known point mutations in the HFE gene. Symptoms and organs involved can vary considerably: Only a small fraction of the 200,000 400,000 persons affected in Germany develop the classical picture of liver cirrhosis and/or pancreatic fibrosis. Nevertheless, the life expectancy of persons with moderate or even subclinical symptoms is reduced, in many cases due to myocardial damage leading to cardiomyopathy with greatly increased risk of sudden cardiac death. Although the high prevalence of HH suggests that sudden cardiac death due to cardiac HH is a relatively common cause of death, the forensic literature lacks such reports. We present the case of sudden cardiac death in a young man with histological findings of massive cardial hemochromatosis which is characterized by the fact that none of the three known mutations for HH were found. This case demonstrates that genetic screening alone might not be sufficient to identify all persons at risk to developing HH. PMID- 15133681 TI - The enhancement of histone H4 and H2A serine 1 phosphorylation during mitosis and S-phase is evolutionarily conserved. AB - Histone phosphorylation has long been associated with condensed mitotic chromatin; however, the functional roles of these modifications are not yet understood. Histones H1 and H3 are highly phosphorylated from late G2 through telophase in many organisms, and have been implicated in chromatin condensation and sister chromatid segregation. However, mutational analyses in yeast and biochemical experiments with Xenopus extracts have demonstrated that phosphorylation of H1 and H3 is not essential for such processes. In this study, we investigated additional histone phosphorylation events that may have redundant functions to H1 and H3 phosphorylation during mitosis. We developed an antibody to H4 and H2A that are phosphorylated at their respective serine 1 (S1) residues and found that H4S1/H2AS1 are highly phosphorylated in the mitotic chromatin of worm, fly, and mammals. Mitotic H4/H2A phosphorylation has similar timing and localization as H3 phosphorylation, and closely correlates with the chromatin condensation events during mitosis. We also detected a lower level of H4/H2A phosphorylation in 5-bromo-2-deoxyuridine-positive S-phase cells, which corroborates earlier studies that identified H4S1 phosphorylation on newly synthesized histones during S-phase. The evolutionarily conserved phosphorylation of H4/H2A during the cell cycle suggests that they may have a dual purpose in chromatin condensation during mitosis and histone deposition during S-phase. PMID- 15133682 TI - Isolated laryngeal lymphangioma showing the symptoms of acute epiglottitis. AB - Lymphangiomas are rare, congenital lesions of the lymphatic system, and about 90% of them are detected by the 2nd year of life. Although the head and neck region is the most common place of presentation, isolated laryngeal lymphangioma is extremely rare. A 37-year-old female patient presented with characteristic symptoms of acute epiglottitis. After her acute symptoms resolved with medical treatment, endoscopy was performed, and a wide, pedunculated mass arising from the epiglottis of the larynx was seen. The mass was totally excised and microscopically diagnosed as lymphangioma. To our knowledge, this is the first such case reported in the literature. In the control examination performed after 9 months, no evidence of recurrence was revealed. This interesting case illustrates that the symptoms of acute epiglottitis in the adult should be further investigated to exclude rare lesions such as lymphangioma. PMID- 15133683 TI - Peripheral giant cell granuloma of the mandibular condyle presenting as a preauricular mass. AB - Preauricular mass is a common symptom for patients presenting to the otorhinolaryngologist with parotid disease. Some rare extraparotid lesions, originating from the temporomandibular joint and the mandible itself, also share the same localization and therefore are to be taken into consideration for the differential diagnosis with parotid lesions. Giant cell granuloma (GCG) was first described by Jaffe in 1953. Peripheral GCG (PGCG) is an exophytic soft tissue lesion originating from the periodontal ligament and periosteum. It is located only within the oral cavity. Central GCG (CGCG) is an uncommon benign fibro osseous lesion generally presenting as an expansible mass with cortical bone defect. It is generally located in the mandible. The brown tumor of hyperparathyroidism and giant cell tumor must be ruled out because of the microscopic similarities of these lesions. The first case of PGCG of the mandible condyle is presented, and attention is drawn to mandibular diseases for the differential diagnosis of the preauricular mass. PMID- 15133684 TI - Vocal risk factors for occupational voice disorders in female teaching students. AB - The objective of the study was to assess voice capabilities and laryngeal abnormalities in female teaching students in order to evaluate risk factors of future occupational voice disorders. One hundred forty-four women (aged 17 to 41 years) were examined using videostroboscopy and voice range profile measurements. Stroboscopically, the subjects were classified into three groups depending on the shape of the glottal closure. Thirty-five phonation-associated alterations and six organic alterations of the vocal folds were found. Subjects with an insufficient glottal closure showed a higher percentage of phonation-associated vocal fold alterations (i.e. vocal nodules) and reached lower maximum sound pressure levels. The results underline the necessity to execute vocal examinations and vocal assessment analyses for candidates of voice intensive professions in order to avoid profession-related dysphonias at a later stage. PMID- 15133685 TI - Effects of surgery on the function of maxillary sinus mucosa. AB - For chronic maxillary sinusitis, the most popular surgical approach for treatment is endoscopic sinus surgery (ESS). After these operations, however, it is not clear how well the surgery restores the normal mucociliary function. We examined the mucociliary clearance (MCC) in maxillary sinuses in chronic sinusitis before ESS and 6 months after the operations. The correlation of histology to MCC was also studied. Measurements of the mean residual mucociliary clearance (MCC) of maxillary sinuses was studied pre- and postoperatively with an isotope method. Biopsies taken during ESS and 6 months postoperatively were studied by electron microscopy (EM). Preoperative residual MCC from the maxillary sinuses was 77+/ 26% (mean+/-SD). Six months postoperatively, residual MCC was 70+/-22%, only slightly better than preoperatively. Residual MCC was considered good (< or = 50%) in 12% of sinuses preoperatively and in 20% postoperatively. In preoperative sinuses, 14% did not show any mucociliary clearance compared to 11% postoperatively. As a single EM finding, metaplasia gave the poorest MCC (91%) and microvilli the best (68%). Mucociliary function in maxillary sinusitis remains poor even 6 months postoperatively, with many pathological findings still visible in the sinus epithelium. MCC correlates well with the histology of the mucosa. Surgery does not significantly improve the mucociliary function of sinus mucosa in chronic maxillary sinusitis. PMID- 15133686 TI - Primary angiosarcoma of the non-irradiated parotid gland: a most uncommon, highly malignant tumor. AB - This case report discusses the clinical presentation, imaging, surgery and further treatment and course of a primary angiosarcoma of a non-irradiated parotid gland. PMID- 15133687 TI - Upper lip swelling caused by a large dentigerous cyst. AB - Swelling of the upper lip can result from various diseases such as salivary tumors, infectious and inflammatory diseases and cysts. Among the latter, dentigerous cysts, typically involving unerupted teeth, are sometimes associated with supernumerary teeth in the maxillary anterior incisors region called the mesiodens. We report an unusual case of a large dentigerous cyst associated with an impacted mesiodens in a 42-year-old male who presented with a slow-growing swelling in the upper lip. PMID- 15133688 TI - Otoacoustic emissions in an aberrant internal carotid artery: a case report. AB - A 52-year-old Korean woman who complained of impaired hearing and tinnitus of the left ear was diagnosed with an aberrant internal carotid artery (ICA) in the middle ear using a temporal bone high resolution CT (HRCT) scan. Upon otoscopic examination, her left eardrum was found to be in contact with a mass in the middle ear cavity. Upon audiometric evaluation, a mild conductive hearing impairment was documented, and a discrepancy in the results of otoacoustic emissions (OAEs) was found for the left ear. This case report studies the aberrant ICA in the middle ear that revealed abnormal OAE findings. PMID- 15133689 TI - The non-use of hearing aids in people aged 75 years and over in the city of Kuopio in Finland. AB - Hearing loss is one of the most prevalent chronic conditions affecting the health of the aged. It is typically medically non-treatable, and hearing aid (HA) use remains the treatment of choice. However, only 15-30% of older adults with hearing impairment possess an HA. Many of them never use it. The purpose of our study was to investigate the use of provided HAs and reasons for the non-use of HAs. This population-based survey was set in the city of Kuopio in eastern Finland. A total of 601 people aged 75 years or older participated in this study. A geriatrician and a trained nurse examined the subjects. Their functional and cognitive capacity was evaluated. A questionnaire about participants' socioeconomic characteristics and the use of HAs were included in the study protocol. The subjects who had an HA were assigned to three groups on the basis of HA use: full-time users, part-time users and non-users. Inquiries were made about the subjective reasons for the non-use of HAs. An HA had been prescribed earlier to 16.6% of the study group. Fourteen percent of the females and 23% of the males had been provided with an HA. The HA owners were older than persons who had not been provided with an HA. Twenty-five percent of the HA owners were non users, and 55% were full-time users. A decline in cognitive or functional capacity and low income explained the non-use of HAs. The most common subjective reasons for the non-use of HAs were that the use did not help at all (10/24), the HA was broken (4/24) or it was too complicated to use (5/24). The non-use of HAs is still common among the aged. Elderly people who have been provided with an HA and who have a cognitive or functional decline are at risk to be a non-user of an HA. Therefore, they need special attention in counseling. PMID- 15133690 TI - Melanotic schwannoma of the neck mimicking a malignant melanoma. AB - We report a case of a 42-year-old female patient who presented with a 5-month history of a right cervical swelling. A fine-needle aspiration biopsy performed elsewhere led to the diagnosis of malignant melanoma. Clinical examination revealed pigmented skin and mucous-membrane lesions of the pharynx. Diagnostic exstirpation of the cervical tumor resulted in the intraoperative diagnosis of a malignant melanoma. Subsequent pathological examination including ultrastructural analysis allowed the revision of the diagnosis to that of melanotic schwannoma, a rare, pigmented nerve sheath tumor. Further analysis of the tissue obtained from neck dissection revealed additional melanotic schwannomas. With regard to prognostic and therapeutic issues, it is necessary to differentiate these tumors from the more common metastatic malignant melanoma. The preoperative differential diagnosis is very difficult, as light microscopy and immunocytochemistry allow no discrimination in small biopsies. Although the occurrence of multiple schwannomas points to a hereditary syndrome, our patient did not fulfill the criteria of Carney's syndrome or other known syndromes, suggesting a so far unknown genetic background. PMID- 15133691 TI - Olfactory dysfunction and daily life. AB - The objective of the present study was to investigate the hypothesis that subjects with parosmia suffer more in their daily life than patients who experience only quantitative olfactory loss. Two hundred five outpatients of the Smell and Taste Clinic and 25 healthy controls were included. The newly developed Questionnaire of Olfactory Disorders (QOD) was administered in combination with other psychometric tests (Beck Depression Inventory, "Befindlichkeitsskala" and the Short Form-36 Health Survey) along with an olfactory test ("Sniffin' Sticks"). Results of the QOD were found to be an appropriate and valid measure of the impact of olfactory dysfunction on daily life. Patients with parosmia and quantitative olfactory dysfunction show higher rates of daily life complaints when compared to patients suffering from quantitative olfactory impairment only (QOD-PS: P=0.005). In addition, hyposmic and anosmic patients indicated significantly more complaints compared to patients with normosmia. Further, female patients seemed to suffer more from olfactory dysfunction than male patients. In conclusion, the assessment of the degree of qualitative olfactory dysfunction may be possible by the use of instruments based on questionnaires regarding daily life problems. PMID- 15133692 TI - On a possible prognostic value of otoacoustic emissions: a study on patients with sudden hearing loss. AB - Otoacoustic emissions (OAE) and pure tone audiogram (PTA) were examined in 26 ears of 25 patients suffering from sudden hearing loss from the 1st day to up to 505 days following the drop of hearing to test the hypothesis whether the OAEs are capable of delivering predictive information about the recovery process. The patients were selected from 50 candidates according to the following criteria: one or both ears exhibited a systematic and significant recovery of pure tone threshold in at least one frequency, OAEs were detectable and PTA available, a conductive hearing loss was excluded, and the auditory brainstem responses (ABR) yielded no signs of retrocochlear disorders. Transitory evoked otoacoustic emissions (TEOAE) and distortion product otoacoustic emissions (DPOAE) were measured under constant stimulus and recording conditions in three to nine sessions. The relation between OAE level and actual pure tone threshold was subject to a regression analysis. The correlation between both parameters is small but significant. Even smaller correlations are observed if the OAE level is related to former hearing loss, whereas the correlation improves if the OAE level is compared to the pure tone threshold measured in a later session. The slopes of individual trajectories that connect the successive results of one ear in a plane defined by hearing loss and OAE level show a remarkable accumulation around zero, i.e., in many cases the OAEs remain unchanged even if the hearing loss decreases. The comparison of the OAE levels measured at an early stage with later audiograms shows that there are only a small number of cases with small initial emissions and good final threshold or large initial emissions and bad final threshold. This means that small initial OAEs end up with a remaining final hearing deficit, whereas a high OAE level immediately after drop of threshold correlates with good outcome. The reliability of an individual prediction based on the OAE level combined with the threshold after sudden hearing loss and the consequences for the physiologic mechanisms underlying the sudden hearing loss remain to be proved in further investigations. PMID- 15133693 TI - Unexplained antepartum fetal deaths: what are the determinants? AB - OBJECTIVE: The objective was to assess fetal, antenatal, and pregnancy determinants of unexplained antepartum fetal death. METHODS: This is a hospital based cohort study of 34,394 births weighing 500 g or more from January 1995 to December 2002. Unexplained fetal deaths were defined as fetal deaths occurring before labor, without evidence of significant fetal, maternal or placental pathology. RESULTS: Ninety-eight unexplained antepartum fetal deaths accounted for 27.2% of 360 total fetal deaths. Two-thirds of these deaths occurred after 36 weeks' gestation. The following factors are independently associated with unexplained fetal deaths: primiparity (OR 1.74; 95% CI 1.21, 2.86); parity of five or more (OR 1.19; 95% CI 1.26, 3.26); low socioeconomic status (OR 1.22; 95% CI 1.14, 2.86); maternal age 40 years or more (OR 3.62; 95% CI 1.22, 4.52); maternal age of 18 years or less (OR 1.79; 95% CI 0.82, 2.89); maternal prepregnancy weight greater than 70 kg (OR 2.20; 95% CI 1.85, 3.68); fewer than three antenatal visits in women whose fetuses died at 31 weeks or more (OR 1.11; 95% CI 1.08, 2.48); birth weight ratio (defined as ratio of birth weight to mean birth weight for gestational age) between 0.85 and 0.94 (OR 1.77; 95% CI 1.28, 4.18) or over 1.45 (OR 2.92; 95% CI 1.75, 3.21); trimester of first antenatal visit. Previous fetal death, previous abortion, cigarette smoking, fetal sex, low maternal weight, fetal-to-placenta weight, and post date pregnancy were not significantly associated with unexplained fetal deaths. CONCLUSION: Several factors were identified that are associated with an increased risk of unexplained fetal deaths. PMID- 15133694 TI - Augmented production of transforming growth factor-beta by cultured peripheral blood mononuclear cells from patients with systemic sclerosis. AB - We sought to determine whether the spontaneous production of transforming growth factor-beta (TGF-beta) by peripheral blood mononuclear cells (PBMC) is increased in patients with systemic sclerosis (SSc). Culture supernatants of PBMC from SSc patients (n = 88) and healthy controls (n = 44) were analyzed by enzyme-linked immunosorbent assay. The production of active TGF-beta1 and total (active and latent) TGF-beta1 by PBMC from patients with limited cutaneous SSc (lSSc) and by PBMC from patients with diffuse cutaneous SSc (dSSc) was significantly elevated compared to the production by PBMC from normal controls. Production of active TGF beta1 by dSSc PBMC was higher than that by lSSc PBMC, although not significantly. Patients with PBMC with increased active or total TGF-beta1 production showed significantly shorter disease duration than patients with PBMC with normal production levels. PBMC from patients without anticentromere antibody showed enhanced active TGF-beta1 production more frequently than those from patients with anticentromere antibody. PBMC from SSc patients more frequently showed enhanced total TGF-beta2 production than PBMC from normal controls. Among each leukocyte subset, spontaneous production of total TGF-beta1 was significantly higher in cultured peripheral monocytes/macrophages, but not in T cells, B cells, or NK cells, from patients than from normal controls. Thus, the enhanced production of TGF-beta by PBMC may contribute to the disease process in SSc PMID- 15133696 TI - Clinical features of the posterior horn tear in the medial meniscus. AB - INTRODUCTION: A lower threshold of suspicion is necessary for the appropriate diagnosis of a posterior horn tear in the medial meniscus. In these cases, radial tears or meniscus detachment from its insertion follow minor trauma and precipitate severe knee pain in middle-aged and elderly patients. The purpose of this paper is to demonstrate the key points for diagnosis through examination of the clinical features of this tear. MATERIALS AND METHODS: Arthroscopic examination of 250 knees with medial meniscus tears (and no ligamentous injuries; over 40 years old) identified 26 knees (26 tears) with a posterior horn tear. Of these 26 tears, 16 were radial, and 10 were detached. RESULTS: Eighty-five percent of patients could recall discrete events that preceded the pain. They described these events as a click or a feeling of shock. Afterwards, most patients complained of severe pain or giving way. Hydrarthrosis involving more than 5 ml was present in 81%. Most radiographs (92%) appeared nearly normal. CONCLUSION: It is important to note that this type of tear of the posterior horn in the medial meniscus is not rare. Because this area is difficult to visualize arthroscopically, it may be overlooked unless the threshold of suspicion is lowered. PMID- 15133697 TI - Surface replacement of the humeral head in rheumatoid arthritis. AB - INTRODUCTION: The concept of a newly developed cup arthroplasty (Durom Cup) involves the replacement of the destroyed joint surface of the humeral head with minimal bone resection. In cases of additional massive cuff tear, the cup can be placed in a more valgic position to articulate with the glenoid and the acromion. The aim of this prospective study was to evaluate the results of this surface replacement as a hemiarthroplasty in rheumatoid arthritis. MATERIAL AND METHODS: Forty-five Durom Cups in 39 patients (30 women, 9 men) with rheumatoid arthritis were evaluated preoperatively and every 3 months postoperatively. Their average age was 62.7+/-12.3 years and the average follow-up 45.1+/-11.6 months with a minimum of 36 months. Concerning the cuff, 15 shoulders had an intact cuff (group A), 18 shoulders a partial tearing or a repaired rotator cuff (group B), and 12 shoulders a massive cuff tear (group C). The Constant Score was used, and the cups were examined radiologically. RESULTS: In group A rheumatic shoulders, the Constant Score increased from 21.5+/-9.6 points preoperatively to 66.1+/-9.8 points at 36 months postoperatively; in shoulders of group B, from 19.6+/-9.7 points preoperatively to 64.9+/-9.6 points at 36 months postoperatively; and in shoulders of group C, from 17.5+/-8.7 points to 56.9+/-9.8 points at the latest follow-up examination. All shoulders were pain-free at the latest examination. No complications, component loosening or changes of cup position were observed. CONCLUSION: The results of the Durom Cup are encouraging. In shoulders with additional massive cuff tear, the limited goal criteria were always achieved. Therefore, cup arthroplasty is a good alternative to other kinds of shoulder endoprostheses in rheumatic shoulders with and without massive cuff tear. PMID- 15133698 TI - Expression of the proto-oncogene c-KIT in normal and tumor tissues from colorectal carcinoma patients. AB - BACKGROUND AND AIMS: The proto-oncogene c-KIT encodes a tyrosine kinase receptor essential during embryonic development and postnatal life. Although deregulated expression of c-KIT has been reported, its role in colorectal carcinoma remains controversial: some authors have described a correlation between c-KIT expression and colorectal cancer (CRC), while others have failed to detect the receptor in the majority of neoplasia examined. To address this question, we designed a prospective study to analyze the expression of c-KIT in normal and neoplastic colonic mucosa of the same patient. PATIENTS AND METHODS: We analyzed the tissues of 20 patients undergoing surgical resection for colorectal carcinoma by reverse transcriptase-polymerase chain reaction, Western blot and immunohistochemistry, whose results were correlated with histopathological parameters. RESULTS: Most patients (90%) showed c-KIT expression in normal tissue both at RNA and protein level, while in neoplastic tissue it was observed in 30% of patients at RNA level and in 10% at protein level. By immunohistochemistry the localization of c-KIT protein in the normal colon was restricted to interstitial cells scattered in the stroma, whereas the non-neoplastic epithelium was always negative. The mucinous carcinomas were all c-KIT negative, whereas the only case in which c-KIT was displayed in the neoplastic epithelium was a G3 adenocarcinoma. CONCLUSION: Most colorectal carcinomas do not express c-KIT. We suggest that c-KIT expression is rarely present in this neoplasia; thus, the use of receptor inhibitors should be conducted in selected sub-groups of colon carcinoma patients, subsequent to the clear demonstration of c-KIT overexpression in the neoplastic cells. PMID- 15133699 TI - Surgical risk factors for Hickman catheter sepsis: a prospective study. AB - Catheter-related sepsis (CRS) is a major cause of morbidity in patients receiving chemotherapy and prolonged parenteral nutrition. To determine whether avoiding emergency insertions by using a planned elective list and adopting a 'no-touch' technique has a role in reducing CRS, all cuffed central venous catheters inserted by the open method between 1999 and 2000 were prospectively followed for a total duration of 12 months. The incidence of early sepsis (within 30 catheter days) that could be attributed to surgical factors was studied. CRS was defined as the presence of any two of the following: (1) signs of clinical sepsis without an obvious focus; (2) positive cultures in blood obtained from the catheter; and (3) clinical improvement following removal. A total of 146 catheters were inserted in 130 patients; 15 had a second and 1 had a third catheter inserted. Early CRS was encountered in 13 cases (9%); 95 catheters were inserted on an elective list and 51 on an emergency basis. The distributions of age, sex, number of lumens, neutrophil count, and underlying diagnosis were similar between the groups. There was no significant difference (P = 1) between elective (9/95) and emergency (4/51) insertions. A total of 47 catheters were inserted by the 'no touch' technique and 48 by the manual technique. There was no significant difference in early sepsis (P = 0.7) between the two techniques (6/47 vs 3/48). Thus avoiding emergency insertion or adopting a 'no-touch' technique does not reduce early CRS. Larger prospective studies are warranted to identify surgical risk factors. PMID- 15133700 TI - Immunohistology of aquaporin-1 and stem cell factor-receptor in human undescended testes. AB - OBJECTIVES: Both aquaporin (AQP) 1 and the stem-cell factor/C-kit system seem to have a definite role in testis function, but very few studies have been reported in humans, especially in the paediatric age group. With the present study we wanted to investigate the expression of these proteins to better delineate their role in normal and pathologic testes. METHODS: Immunohistology using AQP 1 and C kit antibodies was performed on paraffin sections of open-testicular biopsies from 32 undescended testes. The testes of cryptorchid patients, with ages ranging from 2 to 15 years, were biopsied during an orchidopexy operation, after obtaining informed consent. Control biopsies, from 8 patients of matched age, were obtained during operations for inguinal hernia or hydrocele, always after obtaining informed consent. Positive results were recorded as diffuse or focal patterns and scored as weak, moderate or strong immunostaining. RESULTS: AQP 1 antibody strongly depicted microvessel endothelial cells, but was unlabeled in endotubular and interstitial cell lines, in both control and undescended testes. The C-kit immunostaining in normal testes revealed a diffuse, strong staining in the cytoplasm of spermatogonia and primary spermatocytes. However, in the undescended testes a focal C-kit immunolabelling was weakly recognized in both spermatogonial and immature Sertoli cells. CONCLUSIONS: These results indicate a direct involvement of AQP 1 in the regulation of fluid transport across the endothelial cell membranes of testicular microvessels. A role of the C-kit receptor protein is also substantiated by its strong expression in the maturing spermatogonia of the normal testes, but was minimally or not recognizable in undescended prepubertal testes. PMID- 15133701 TI - Intraoperative electrical stimulation mapping as an aid for surgery of intracranial lesions involving motor areas in children. AB - OBJECTIVE: We analysed the usefulness of intraoperative electrical stimulation mapping (ESM) for locating motor pathways in pediatric patients harboring cerebral lesions closely related to motor areas. METHODS: We applied ESM in 17 consecutive pediatric patients operated on under general anesthesia. It was possible to locate motor function in 15 patients and in all children 5 years old and younger, as well as in all patients presenting with severe motor deficits, using relatively high current intensities. Intraoperative seizures occurred in 20% of our patients. A macroscopically complete removal of the lesion was carried out in 12 cases out of 17 with no definitive postoperative aggravation. Motor function improved for all patients presenting preoperatively with a severe paresis. CONCLUSION: In our experience ESM revealed to be an useful tool for allowing us to push the resection of any lesion infringing on eloquent cortex up to the limit of functional areas, even in cases in very young and severely paretic children. PMID- 15133702 TI - Complications of ventriculosubgaleal shunts in infants and children. AB - INTRODUCTION: The ventriculosubgaleal shunt has been used for the temporary bypass of the normal cerebrospinal fluid (CSF) pathways. To date, a large series of complications from this procedure has not been elaborated upon in the literature. PATIENTS AND METHODS: We retrospectively reviewed all such shunts (170) placed at our institution over the last 6 years and documented all complications from this procedure. The majority of patients operated upon were premature infants with intraventricular hemorrhage and subsequent hydrocephalus. This technique was used in a much smaller group of patients in whom the peritoneal cavities were not currently candidates for distal shunt implantation but would have been with time. Other patients in whom this technique was used were those with malignant brain tumors, intraventricular abscesses, chronic truncal wounds, chronic subdural hygromas, and meningitis. RESULTS: Complications from subgaleal shunting included infection (5.9%), intracranial hemorrhage (1.1%), and wound leakage (4.7%). CONCLUSIONS: We believe the benefits afforded by ventriculosubgaleal shunting significantly outweigh the risks of the procedure and greatly ease the burden of care for this select population of children. Based on the literature and our own experience, the complications from this procedure are not excessive or extraordinarily unique compared with other neurosurgical CSF diversion techniques. PMID- 15133703 TI - Isolated atlantal stenosis in a patient with idiopathic growth hormone deficiency, and Klippel-Feil and Duane's syndromes. AB - OBJECTIVE AND IMPORTANCE: Isolated atlantal stenosis is very rare with less than ten cases reported, mostly in adult Asians; however, this pathology should be recognized by the neurosurgeon. An unusual case of this rare clinical entity is reported in conjunction with multiple other anomalies. CLINICAL PRESENTATION: We report a Caucasian girl with symptomatic stenosis of the first cervical vertebrae who presented with episodes of loss of tone with subsequent falling, facial cyanosis, urinary incontinence, hand weakness, and difficulties with swallowing. This patient also had the diagnoses of Duane's and Klippel-Feil syndromes, and idiopathic growth hormone deficiency. In addition, this patient was found to have retroflexion of the odontoid process, which further compromised the spinal canal. Flexion-extension radiographs failed to identify cervical spine instability. INTERVENTION: Following suboccipital craniectomy and the removal of the posterior arch of the atlas, the patient's symptoms were resolved and her urinary incontinence improved. CONCLUSIONS: We theorize that our case represents a variant of Wildervanck syndrome. Further case reports are necessary to verify that isolated atlantal stenosis is a component of this constellation of findings. Furthermore, our case demonstrates that degenerative changes of the spine are not necessary for the production of symptoms in cases of isolated stenosis of the atlas. PMID- 15133704 TI - Treatment and management of the Chiari II malformation: an evidence-based review of the literature. AB - OBJECTIVE: Multiple surgical strategies exist for the management of the symptomatic Chiari II malformation. To date, no comprehensive analysis of this medical literature in an attempt to seek out standards or guidelines has been performed, thus serving as the impetus for this present review. METHODS: A computerized search of the database of the National Library of Medicine was performed on the English-language medical literature between 1966 and 2003. Terms searched for included hindbrain hernia, Chiari II, Arnold-Chiari, surgery, decompression, syringomyelia, and CSF shunts. All literature found was reviewed with associated references from these sources. Each piece of literature reviewed was scrutinized and guidelines developed utilizing the methodology used by the Guidelines for the Management of Acute Cervical Spine and Spinal Cord Injuries. RESULTS: All data reviewed were deemed to be Class III in nature. No standards or guidelines could be established from the available literature. There is significant debate and variable results in the current neurosurgical literature regarding the evaluation of shunt function vs. Chiari II decompression in patients with symptomatic hindbrain herniation. CONCLUSIONS: Multi-institutional prospective randomized trials are needed before any conclusions can be drawn regarding symptomatic Chiari II and its treatment paradigm. Until this time, clinical practice will be based on individual surgeons' experience, training, and personal beliefs. PMID- 15133705 TI - The steady-state force-Ca2+ relationship in intact lobster (Homarus americanus) cardiac muscle. AB - The heart of the decapod crustacean is activated by regular impulse bursts from the cardiac ganglion. The cardiac pump function depends on ganglionic burst frequency, burst duration, and burst impulse frequency. Here, we activated isolated lobster cardiac ostial muscle (Orbicularis ostii muscle, OOM) by stimulus trains in vitro in order to characterize the response of the contractile apparatus to [Ca2+]i. We employed stimulus trains that generate a steady state between the [Ca2+]i and force in order to estimate the Ca2+ sensitivity of myofilaments. Force and [Ca2+]i transients were simultaneously recorded using a silicon strain gauge and the fluorescence of iontophoretically microinjected fura 2 salt. We examined the effects of tetanus duration (TD), the interval between trains, and 6 microM cyclopiazonic acid, an inhibitor of the SR Ca2+ pump, on the steady-state force-[Ca2+]i relationship. The instantaneous force-[Ca2+]i relationships appeared sigmoidal (EC50 and Hill coefficient, 98.8+/-32.7 nM and 2.47+/-0.20, mean +/- SD, respectively), as did the curves superimposed after 500 ms following the start of stimulation, indicating that the force-[Ca2+]i relationship had reached a steady state at that time. Also, the maximum activated force (Fmax) was estimated using the steady-state force-[Ca2+]i relationship. Prolonged stimulus trains, decreasing the interval between recurrent trains from 5 to 2.5 s, and cyclopiazonic acid each increased the measured EC50 without changing Fmax. The EC50 correlated strongly with averaged [Ca2+]i over time. We conclude that the steady-state force-[Ca2+]i relationships in the OOM indicate cooperation between force generation and Ca2+ binding by the myofilaments. Our data also suggest the existence of a novel Ca2+-dependent mechanism which reduces Ca2+ sensitivity and accelerates relaxation of lobster cardiac muscle myofilaments. PMID- 15133706 TI - [Malignant glaucoma following blunt trauma of the eye]. AB - PURPOSE: Ciliary block glaucoma most commonly occurs after intraocular surgery. We report a case of malignant glaucoma following blunt trauma of the eye with no previous surgery or laser. PATIENT: A 54-year-old female suffered from acute elevation of intraocular pressure on her right eye after blunt trauma 2 months earlier. As a variety of topical and systemic antiglaucomatous drugs and the presence of a patent iridotomy could not control intraocular pressure (IOP) and the anterior chamber remained flattened, ciliary block glaucoma was diagnosed. Subsequent surgical treatment included pars plana vitrectomy (PPV), phacoemulsification with implantation of an intraocular lens, a second PPV with trabeculectomy and mitomycin-C, eccentric YAG-laser capsulotomy, and a third PPV with zonulo-iridectomy and endolaser-induced zonular shrinkage. After this variety of surgical approaches, a stable and deep anterior chamber could be achieved with an average IOP of 10 mmHg. CONCLUSIONS: Ciliary block glaucoma may occur after blunt trauma of the eye even without a history of intraocular surgery or laser. In addition, our case demonstrates the uncertainty of surgical success in affected patients. PMID- 15133707 TI - [Triple surgery. Keratoplasty combined with cataract extraction and intraocular lens implantation]. AB - Since the 1980s, combined perforating keratoplasty and extracapsular cataract surgery with intraocular lens implantation (triple procedure) has been performed with increasing success for patients with corneal disease and cataract. Advantages include prompt visual rehabilitation, protection of the transplanted endothelium from sequential cataract surgery, and similar up to superior functional results following triple procedures. In the following review article, the authors discuss advantages and disadvantages of triple procedures, alternative surgical techniques, problems in intraocular lens power calculation, intraoperative pitfalls, and postoperative results of combined keratoplasty, cataract removal, and intraocular lens implantation. PMID- 15133708 TI - [Stability of postoperative visual acuity after rt-PA -SF6-treatment of submacular hemorrhage in age-related macular degeneration]. AB - PURPOSE: Treatment of submacular hemorrhage with recombinant tissue plasminogen activator(rt-PA) and SF6 induces blood displacement out of the fovea by maintaining prone positioning. The underlying disease, e.g. the choroidal neovascularisation remains untreated. Nevertheless, it has been noted that the visual results of this method are more favorable than those expected in the natural course. Contributing long-term results the present study intends to give more information concerning this phenomenon. METHOD: A total of 45 consecutive eyes were treated using a standardized procedure (duration of hemorrhage < or =1 week, area of hemorrhage 1-10 disc areas, 50 gammag rt-PA, 0.5 ml SF6 24 h later, prone positioning for 3 days). Those with a follow-up of > or =6 months were included in the study. RESULTS: From 25 patients a total of 25 eyes fulfilled the criteria. Postoperative visual acuity(3-7 days) was improved or unchanged in 19 eyes and worse in 6 eyes. Comparison of postoperative visual acuity after follow up of an average of 14.2 months (range 6-28 months) revealed an improvement in 10 eyes, no change in 7 eyes and a deterioration in 8 eyes. A second treatment was performed in 2 eyes and photocoagulation in 1 eye. CONCLUSION: After more than 1 year the postoperative visual acuity was stable or even better in about two thirds of the eyes therefore treatment with rt-PA and SF6 may also influence the choroidal neovascularisation. PMID- 15133709 TI - [Follow-up examination of eyes with chronic open-angle glaucoma and optic disc hemorrhages]. AB - PURPOSE: To evaluate the frequency of neuroretinal rim loss in glaucomatous eyes with ophthalmoscopically detected optic disc hemorrhages. METHODS: The prospective comparative clinical observational study included 78 eyes from 69 Caucasian patients with chronic open-angle glaucoma and a flame-shaped optic disc hemorrhage at the time of presentation, and 386 eyes from 252 patients with chronic open-angle glaucoma without disc hemorrhages. All patients underwent repeated qualitative and morphometric evaluation of color stereo optic disc photographs. RESULTS: Patients with disc hemorrhages were older than patients without hemorrhages and showed an initially smaller neuroretinal rim area. Of the 78 eyes with disc hemorrhages 39 showed loss of neuroretinal rim during the follow-up period. For the remaining 39 eyes, no changes of the neuroretinal rim could be detected on optic disc photographs. Of the 386 eyes without disc hemorrhages 71 showed loss of neuroretinal rim during the follow-up period. A survival analysis confirmed a hazard ratio of three between eyes with and without disc hemorrhages and a hazard ratio of 1.85 per decade of patient's age (multivariate analysis). CONCLUSIONS: Disc hemorrhages lead to a 3-fold increase of risk for further retinal rim loss in eyes with chronic open-angle glaucoma. PMID- 15133710 TI - Virtual endoscopy of the tracheo-bronchial system: sub-millimeter collimation with the 16-row multidetector scanner. AB - The purpose of this study was to evaluate the scope of sub-millimeter collimation reconstruction parameters using 16-row computer tomography and ECG triggering on image quality in virtual bronchoscopy. Thirty-two patients (5 women, 27 men, mean age 66.6+/-1.4) who had been admitted for coronary artery bypass graft surgery underwent CT examination of the thorax (Sensation 16, Siemens, Inc., Forchheim, Germany). All patients were examined with 16x0.75-mm collimation. Image reconstruction was performed for two groups. In group A ( n=32), slice thickness of 1.5 mm and an overlap of 0.75 mm were used. In group B ( n=32), slice thickness of 0.75 mm and an overlap of 0.4 mm were applied. Retrospective ECG triggering was performed in all patients. The maximum order of recognizable bronchi was determined in each data set. In addition to assessing the maximum order of bronchial bifurcation, bronchial diameter was determined in truly perpendicular sections in each patient. For every segment proximal to a bifurcation, image quality was subjectively graded as poor (grade 1), moderate (grade 2) or good (grade 3). The observers were asked to identify the minimum cardiac movement ECG-triggered image sets assuming that they would be of better quality than the maximum cardiac movement ECG-triggered image sets. The Mann Whitney U-test and the Fisher's Exact Test were used for statistical evaluation. In group A, a mean of 4.8+/-0.2 bifurcations was ascertained vs. 6.5+/-0.3 bifurcations in group B [ P<0.0003]. For bronchial diameters in group A, a mean of 7.5+/-0.4 mm was determined vs. 4.6+/-0.4 mm in group B [ P<0.0001]. In group B, two independent radiologists observed a significant shift to better image quality in all segments evaluated [ P<0.006 to P<0.000001]. Motion artifacts were judged as being significantly reduced by minimum cardiac movement ECG-triggering in group B [observer 1: P=0.0007 (20/32); observer 2: P=0.008 (18/32)], but not in group A [observer 1: P=0.286 (13/32); observer 2: P=0.123 (16/32)]. Sub millimeter collimation and minimum cardiac movement ECG-triggered data acquisition allow deeper penetration into the tracheo-bronchial system allowing visualization of the bronchial surface down to diameters below 5 mm in certain cases up to the eighth bifurcation. Along with an enhanced visualization as such, better image quality is acquired in all segments evaluated. Trade off between better image quality, of doubtful diagnostic consequence, and much higher irradiation dose must be made. PMID- 15133711 TI - Cytokinin-induced abnormal shoot organogenesis is associated with elevated Knotted1-type homeobox gene expression in tobacco. AB - The molecular mechanisms that regulate the transcription of key developmental genes involved in shoot organogenesis have yet to be fully elucidated. However, it is clear that plant growth regulators, such as cytokinin, play a critical role in the differentiation of adventitious shoots. In Nicotiana tabacum zz100 leaf discs, high frequency shoot formation could be induced with 5 microM of the cytokinin N6-benzyladenine (BA). Increasing the exogenous BA concentration to greater than 20 microM resulted in stunted explants with abnormal shoot morphology and altered mineral composition. Explants with abnormal shoots did not appear to be hyperhydric. Abnormalities were, however, associated with an increase in the expression of a knotted1-type homeobox gene (TobH1) isolated from normal shoot-forming cultures. The results suggest that the development of cytokinin-induced abnormal shoot morphology possibly involves changes in TobH1 gene expression. PMID- 15133712 TI - Agrobacterium-mediated genetic transformation and development of herbicide resistant sugarcane (Saccharum species hybrids) using axillary buds. AB - Direct regeneration from explants without an intervening callus phase has several advantages, including production of true type progenies. Axillary bud explants from 6-month-old sugarcane cultivars Co92061 and Co671 were co-cultivated with Agrobacterium strains LBA4404 and EHA105 that harboured a binary vector pGA492 carrying neomycin phosphotransferase II, phosphinothricin acetyltransferase (bar) and an intron containing beta-glucuronidase (gus-intron) genes in the T-DNA region. A comparison of kanamycin, geneticin and phosphinothricin (PPT) selection showed that PPT (5.0 mg l(-1)) was the most effective selection agent for axillary bud transformation. Repeated proliferation of shoots in the selection medium eliminated chimeric transformants. Transgenic plants were generated in three different steps: (1) production of putative primary transgenic shoots in Murashige-Skoog (MS) liquid medium with 3.0 mg l(-1) 6-benzyladenine (BA) and 5.0 mg l(-1) PPT, (2) production of secondary transgenic shoots from the primary transgenic shoots by growing them in MS liquid medium with 2.0 mg l(-1) BA, 1.0 mg l(-1) kinetin (Kin), 0.5 mg l(-1) alpha-napthaleneacetic acid (NAA) and 5.0 mg l(-1) PPT for 3 weeks, followed by five more cycles of shoot proliferation and selection under same conditions, and (3) rooting of transgenic shoots on half strength MS liquid medium with 0.5 mg l(-1) NAA and 5.0 mg l(-1) PPT. About 90% of the regenerated shoots rooted and 80% of them survived during acclimatisation in greenhouse. Transformation was confirmed by a histochemical beta-glucuronidase (GUS) assay and PCR amplification of the bar gene. Southern blot analysis indicated integration of the bar gene in two genomic locations in the majority of transformants. Transformation efficiency was influenced by the co-cultivation period, addition of the phenolic compound acetosyringone and the Agrobacterium strain. A 3-day co-cultivation with 50 micro M acetosyringone considerably increased the transformation efficiency. Agrobacterium strain EHA105 was more effective, producing twice the number of transgenic shoots than strain LBA4404 in both Co92061 and Co671 cultivars. Depending on the variety, 50-60% of the transgenic plants sprayed with BASTA (60 g l(-1) glufosinate) grew without any herbicide damage under greenhouse conditions. These results show that, with this protocol, generation and multiplication of transgenic shoots can be achieved in about 5 months with transformation efficiencies as high as 50%. PMID- 15133713 TI - Radiographic hip osteoarthritis and acetabular dysplasia in Turkish men and women. AB - The prevalence of hip osteoarthritis (OA) varies with different regions, and the reasons for this remain unclear. Mild acetabular dysplasia has been considered a risk factor for hip OA, but recent studies have not confirmed this. In Turkey, the prevalence of significant radiographic hip OA, i.e. Kellgren-Lawrence (K-L) grades 3 and 4, is lower than in Western populations. We aimed to investigate the potential effects of radiographic measures of acetabular dysplasia on this discrepancy. Ninety-two Turkish patients (65 males, 27 females) aged 55 and over were studied. Plain supine abdominal radiographs and intravenous pyelographs were evaluated. Centre-edge (CE) angle and acetabular depth (AD) of each hip were measured, and K-L grading was done. The mean CE angles +/- SD of right and left hips were 34+/-7 degrees and 35+/-7 degrees (range 14-52 and 18-50), respectively. The mean AD+/-SD of right and left hips was 13.7+/-3.4 and 13.6+/ 3.4 (range 6-23 and 8-25), respectively. Mean CE angle or acetabular depth of the hips with K-L grade 2 did not differ from those with grades 0 and 1. There was no patient with grade 3 or 4. The prevalence of acetabular dysplasia based on a CE angle of <25 degrees was 9.8% in both right and left hips. When it was defined as having an AD of <9 mm, the prevalence was 4.3% and 1.1% in the right and left hips, respectively. Overall, 10.4% of the hips (13% of the men and 3.7% of the women) had acetabular dysplasia based on at least one of these measurements. Mild acetabular dysplasia appears to be relatively common among Turks aged 55 and over. However, it does not seem to play a major role in the development of radiographic hip OA. PMID- 15133714 TI - A gene of the opsin family in the carotenoid gene cluster of Fusarium fujikuroi. AB - Opsins are membrane photoreceptors closely related to the heat-shock proteins of the HSP30 family. Their functions include light-driven ion pumping in archaea and light detection in algae and animals, using the apocarotenoid retinal as a light absorbing prosthetic group. We describe a gene of Fusarium fujikuroi, carO, coding for a polypeptide resembling opsins and HSP30-like proteins and contiguous to the genes of the carotenoid pathway, carRA and carB. Transcription of carO is induced by light and is deregulated in carotenoid-overproducing mutants. The same regulation pattern is exhibited by carRA and carB; and common conserved DNA elements are found in the three promoters. Heat shock resulted in a modest induction of carO transcription, similar to the one exhibited by carB, confirming a common regulation. Targeted mutagenesis of carO produced no apparent phenotypic modification, including no change in the photoinduction of carotenoid biosynthesis. PMID- 15133715 TI - A thermostable lipase produced by a newly isolated Geotrichum-like strain, R59. AB - Growth and production of lipase by a new Geotrichum-like strain, R59, were studied. Production of extracellular lipase was substantially enhanced when the initial pH of the culture medium, types of carbon and nitrogen sources, substances probably stimulating the lipase biosynthesis, the temperature, and time of growth were optimized. Sucrose and triolein were the most effective carbon sources for lipase production. Maximum lipase activity (146 U/ml(-1)) was obtained with urea as the nitrogen source. Growth at 30 degrees C, an initial pH of 6.0 and incubation time of 48 h were found as optimum conditions for cell growth and production of lipase by Geotrichum-like strain R59. The enzyme was thermostable and exhibited very high activity after 1 h incubation at 60 degrees C. PMID- 15133716 TI - Glutamate as an inhibitor of phosphoenolpyruvate carboxylase activity in Corynebacterium glutamicum. AB - The glutamate-producing bacterium, Corynebacterium glutamicum is known to possess two anaplerotic enzymes: pyruvate carboxylase (Pc) and phosphoenolpyruvate carboxylase (PEPc). In vitro, this latter enzyme appeared to be inhibited by different glutamic acid salts, whereas ammonium-glutamate had no influence on Pc activity. To investigate the in vivo relevance of PEPc activity inhibition, the intracellular concentration of glutamate was determined throughout the glutamate producing process. The intracellular concentration was then shown to be sufficient to induce a dramatic inhibition of PEPc activity during the process. As a consequence, intracellular accumulation of glutamate could be at least partially responsible for the weak participation of PEPc within the anaplerosis activity in amino-acid-producing strains of C. glutamicum. PMID- 15133717 TI - In silico physical mapping software for the Triticum aestivum genome. AB - The large size of the Triticum aestivum genome makes it unlikely that a complete genome sequence for wheat will be available in the near future. Exploiting the conserved genome organization between wheat and rice and existing genomic resources, we have constructed in silico physical mapping software for wheat, assigning a gross physical location(s) into chromosome bins to 22,626 representative wheat gene sequences. To validate the predictions from the software we compared the predicted locations of ten ESTs to their positions experimentally determined by SNP marker analysis. Six of the sequences were correctly positioned on the map including four that demonstrated a high level of colinearity with their orthologous rice genomic region. This tool will facilitate the development of molecular markers for regions of interest and the creation of map-based cloning strategies in areas demonstrating high levels of sequence conservation and organization between wheat and rice. PMID- 15133718 TI - A novel missense ATP1A2 mutation in a Finnish family with familial hemiplegic migraine type 2. AB - Familial hemiplegic migraine (FHM), a rare autosomal dominant subtype of migraine with aura, has been linked to two chromosomal loci, 19p13 and 1q23. Mutations in the Na+K+-ATPase alpha2 subunit gene, ATP1A2, on 1q23 have recently been shown to cause familial hemiplegic migraine type 2 (FHM2). We sequenced the coding regions of this gene in a Finnish chromosome 1q23-linked FHM family with associated symptoms such as coma and identified a novel A1033G mutation in exon 9. This mutation results in a threonine-to-alanine substitution at codon 345. This residue is located in a highly conserved N-terminal region of the M4-5 loop of the Na+,K+-ATPase. Furthermore, the T345A mutation co-segregated with the disorder in our family and was not present in 132 healthy Finnish control individuals. For these reasons it is most likely the FHM-causing mutation in this family. PMID- 15133719 TI - The estrogen receptor 1 G594A polymorphism is associated with migraine susceptibility in two independent case/control groups. AB - Migraine is a painful and debilitating disorder with a significant genetic component. Steroid hormones, in particular estrogen, have long been considered to play a role in migraine, as variations in hormone levels are associated with migraine onset in many sufferers of the disorder. Steroid hormones mediate their activity via hormone receptors, which have a wide tissue distribution. Estrogen receptors have been localized to the brain in regions considered to be involved in migraine pathogenesis. Hence it is possible that genetic variation in the estrogen receptor gene may play a role in migraine susceptibility. This study thus examined the estrogen receptor 1 (ESRalpha) gene for a potential role in migraine pathogenesis and susceptibility. A population-based cohort of 224 migraine sufferers and 224 matched controls were genotyped for the G594A polymorphism located in exon 8 of the ESR1 gene. Statistical analysis indicated a significant difference between migraineurs and non-migraineurs in both the allele frequencies (P=0.003) and genotype distributions (P=0.008) in this sample. An independent follow-up study was then undertaken using this marker in an additional population-based cohort of 260 migraine sufferers and 260 matched controls. This resulted in a significant association between the two groups with regard to allele frequencies (P=8 x 10(-6)) and genotype distributions (P=4 x 10( 5)). Our findings support the hypothesis that genetic variation in hormone receptors, in particular the ESR1 gene, may play a role in migraine. PMID- 15133721 TI - A review of treatment interventions in whiplash-associated disorders. AB - In recent years, there has been much debate on the treatment of whiplash associated disorders (WAD). It is not clear if the treatments commonly employed are effective, and concerns have been raised on the available scientific evidence of many of these treatments. The aim of this study was to review the literature systematically to analyze the evidence basis of many commonly used treatments for patients suffering from WAD, both in the acute and the chronic state. A computer assisted search of the databases Medline (from 1962 to May 2003), CINAHL (1960 2003), Embase (1976-2003), and Psychinfo (1960-2003) was conducted as well as a check of the reference lists of relevant studies. All randomized controlled trials (RCTs) were retrieved and systematically analyzed with three common instruments of measurement of methodological quality. A qualitative analysis ("best-evidence synthesis") was performed. The methodological quality of 26 RCTs was analyzed. The median quality scores for all three instruments were poor. Based on the degrees of evidence and the practical obstacles, the following treatments can be recommended: Early physical activity in acute WAD, radiofrequency neurotomy, combination of cognitive behavioral therapy with physical therapy interventions, and coordination exercise therapy in chronic WAD. High-quality RCTs are not common in the field of WAD. More research is needed, particularly on the treatment of chronic WAD. PMID- 15133722 TI - What is the evidence for using hemostatic agents in surgery? AB - The pharmacological methods used to achieve systemic hemostasis have generated much discussion due to concerns of serious adverse effects (e.g., thromboembolic complications) and costs of therapy in addition to efficacy considerations. There are a limited number of well-controlled trials involving pharmacological hemostasis for spine surgery. In the largest double-blinded randomized controlled trial to date involving spine surgery, there was a trend toward reduced homologous transfusion in patients receiving aprotinin, but the only statistically significant result ( p<0.001) was a reduction in autologous red cell donations. The findings of this trial are important, since the investigators used a number of restrictive transfusion strategies (e.g., autologous donation, low hematocrit trigger for transfusion, blood-salvaging procedures with the exception of no cell saver) that were not always employed in earlier trials involving hemostatic agents. Smaller studies involving antifibrinolytic agents other than aprotinin have demonstrated reductions in blood loss and transfusion requirements in patients undergoing spine surgery, although the results were not always statistically significant. A very large randomized trial would be required to address comparative medication- and transfusion-related adverse events; such a trial involving patients undergoing cardiac surgery is currently being performed. Additionally, cost-effectiveness analyses are needed to help define the role of these agents based on the data that is available. PMID- 15133723 TI - Kinematic response of lumbar functional spinal units to axial torsion with and without superimposed compression and flexion/extension. AB - Experimental data suggest that lumbar torsion contributes to lumbar disc degenerative changes, such as instability, spondylolisthesis and spinal canal stenosis. However, some basic mechanical characteristics of the lumbar spine under torsional loading have not yet been reported in detail. For example, the function of the facet joints under combined mechanical loads such as torsion with superimposed flexion or extension postures is an area of interest about which little biomechanical data have been reported. In this study, the kinematic response to axial torsion with superimposed axial compression (200 N), compression-flexion (3 and 6 Nm) and compression-extension (3 and 6 Nm) was investigated in 10 cadaveric lumbar functional spinal units. Range of motion (ROM), and helical axes of motion (HAM), were analyzed. There was no difference in ROM between no preload, pure compressive and flexion-compression preload conditions. The ROM was significantly reduced by both extension-compression preload conditions (11% reduction for 3 Nm and 19% reduction for 6 Nm of extension) compared to the pure compressive preload. For no preload, the average HAM position in the transverse plane of the intervertebral disc was near the posteriormost part of the disc and located laterally on the side contralateral to the applied torsional moment. In the transverse plane, the HAM position showed a discrete trend towards the posterior part of the specimens during extension. Kinematic data were visualized using computer animation techniques and CT-based reconstructions of the respective specimens. This information may be used for identifying and characterizing physiologic and pathologic motion and for specifying conservative and surgical treatment concepts and, thus, may find application to identifying indications for spinal fusion or in evaluating the effect of future semi-flexible instrumentation. PMID- 15133724 TI - Weed control and cover crop management affect mycorrhizal colonization of grapevine roots and arbuscular mycorrhizal fungal spore populations in a California vineyard. AB - Arbuscular mycorrhizal (AM) fungi naturally colonize grapevines in California vineyards. Weed control and cover cropping may affect AM fungi directly, through destruction of extraradical hyphae by soil disruption, or indirectly, through effects on populations of mycorrhizal weeds and cover crops. We examined the effects of weed control (cultivation, post-emergence herbicides, pre-emergence herbicides) and cover crops (Secale cereale cv. Merced rye, x Triticosecale cv.Trios 102) on AM fungi in a Central Coast vineyard. Seasonal changes in grapevine mycorrhizal colonization differed among weed control treatments, but did not correspond with seasonal changes in total weed frequency. Differences in grapevine colonization among weed control treatments may be due to differences in mycorrhizal status and/or AM fungal species composition among dominant weed species. Cover crops had no effect on grapevine mycorrhizal colonization, despite higher spring spore populations in cover cropped middles compared to bare middles. Cover crops were mycorrhizal and shared four AM fungal species (Glomus aggregatum, G. etunicatum, G. mosseae, G. scintillans) in common with grapevines. Lack of contact between grapevine roots and cover crop roots may have prevented grapevines from accessing higher spore populations in the middles. PMID- 15133725 TI - De Toni-Debre-Fanconi syndrome due to a palindrome-flanked deletion in mitochondrial DNA. AB - We report the molecular findings in a child presenting with sideroblastic anemia and proximal tubulopathy. Analysis of mitochondrial DNA (mtDNA) from fibroblasts showed the presence of a 3.3-kb single deletion in 50% of the genomes. This mutation is, unlike other previously reported deletions in tubulopathy patients, not flanked by direct repeat sequences but by palindrome sequences at the deletion breakpoints, suggesting an unusual mechanism for production of deletion. These findings further expand our knowledge of the syndrome of anemia and tubulopathy due to single deletions of mtDNA. PMID- 15133726 TI - Treatment of Wilms tumor-related hypertension with losartan and captopril. AB - Hypertension is commonly associated with Wilms tumor, but hypertension secondary to renin-secreting Wilms tumor is uncommon. We present an infant with severe hypertension and renin hypersecretion, which was resistant to multiple antihypertensive agents. Blood pressure was eventually controlled with a combination of captopril and losartan. PMID- 15133727 TI - End-stage renal disease from glomerulonephritis associated with anti-phospholipid syndrome. AB - Primary anti-phospholipid syndrome (APS) is perceived to be an uncommon disorder, infrequently recognized as a cause of renal disease in childhood. While renal involvement in APS classically manifests as thrombotic events, other renal diseases associated with APS have been reported in adults, including membranous nephropathy and minimal change disease. We report our experience of caring for a child who presented with acute anuric renal failure due to anti-neutrophilic cytoplasmic antibody-negative rapidly progressive glomerulonephritis (RPGN), with concomitant thrombotic microangiopathy (TMA). Recognition of the APS as a cause of the patient's TMA facilitated institution of anticoagulation. Our patient's renal failure did not improve and the patient remained dependent on dialysis until he was successfully transplanted. The purpose of our report is to make health-care professionals aware of the previously unreported association of pauci immune RPGN and APS in children; early recognition of APS will allow initiation of anticoagulation to prevent recurrent thromboses and enable successful transplantation. PMID- 15133728 TI - Blood pressure nomograms for school children in Iran. PMID- 15133729 TI - Addressing a whole bioprocess in real-time using an optical biosensor-formation, recovery and purification of antibody fragments from a recombinant E. coli host. AB - The use of biosensor technology is described to address in real-time the production and subsequent purification of a bioactive recombinant protein product. The product, D1.3 Fv antibody fragment, was expressed in Escherichia coli and purified via two process routes, one for extracellular and one for intracellular product material. The cells were harvested by centrifugation in a solid bowl CARR Powerfuge and stored at -70 degrees C. Clarification of the supernatant was performed by depth filtration, followed by affinity chromatography for final purification of the extracellular product. To purify the intracellular product the harvested cells were resuspended and homogenised. Removal of debris in the CARR Powerfuge was followed by depth filtration and affinity chromatography. In this work we have shown the rapid determination of bioactive product levels, and the impact this has on improved accountability and confidence is demonstrated in process mass balances on the product using the data acquired during process operation. PMID- 15133730 TI - Activated sludge encapsulation in gellan gum microbeads for gasoline biodegradation. AB - A two-phase dispersion technique, termed emulsification-internal gelation, is proposed for encapsulation of activated sludge in gellan gum microbeads. The influence of emulsion parameters on size distribution of microbeads was investigated. Mean diameter of microbeads varied within a range of 34-265 microm as a descending function of emulsion stirring rate (1,000-5,000 rpm), emulsification time (10-40 min), and emulsifier concentration (0-0.1% w/w), and as an ascending function of disperse phase volume fraction (0.08-0.25). Encapsulated sludge expressed a high biodegradation activity compared with non encapsulated sludge cultures even at 4.4 times lower level of overall biomass loading. Over 90% of gasoline at an initial concentration of 35 and 70 mg l(-1) was removed by both encapsulated and non-encapsulated sludge cultures in sealed serum bottles within 7 days. Encapsulation of activated sludge in gellan gum microbeads enhanced the biological activity of microbial populations in the removal of gasoline hydrocarbons. The results of this study demonstrated the feasibility of the production of size-controlled gellan gum-encapsulated sludge microbeads and their use in the biodegradation of gasoline. PMID- 15133731 TI - Effects of opportunistic predation on anti-predator behavioural responses in a guild of ground foragers. AB - We studied factors that affect prey selection by a generalist predator that opportunistically attacks prey species, and the associated inter- and intra specific responses of prey to this type of predation. Our model system was a guild of ground-foraging birds that are preyed upon by magpies ( Pica pica) during the breeding season. We found that magpies attacked up to 12 species during three consecutive breeding seasons. The overall capture success was estimated to be 4.9%. Magpies tended to attack from the air, targeting solitary prey, either on the ground or flying. Inter-specific prey responses to the risk of magpie predation included a reduction in the mean number of species occupying a foraging patch when magpies were present and a decrease in the distance between heterospecific neighbours. Intra-specific responses to magpie predation varied between species that were subject to different attack rates. Preferentially attacked prey enhanced their risk responses (increase in scanning time and scanning rate in the presence of magpies) relative to those species attacked in proportion to their abundance (increase only in scanning rate with magpies). Species attacked infrequently, relative to their abundance, showed no antipredator response. The probability of being attacked, rather than mortality rate, appears to be the factor to which prey species respond. PMID- 15133732 TI - Nitrogen uptake and turnover in riparian woody vegetation. AB - The nutrient balance of streams and adjacent riparian ecosystems may be modified by the elimination of anadromous fish runs and perhaps by forest fertilization. To better understand nitrogen (N) dynamics within stream and riparian ecosystems we fertilized two streams and their adjacent riparian corridors in central Idaho. On each stream two nitrogen doses were applied to a swathe approximately 35 m wide centered on the stream. The fertilizer N was enriched in 15N to 18 per thousand. This enrichment is light relative to many previous labeling studies, yet sufficient to yield a traceable signal in riparian and stream biota. This paper reports pre-treatment differences in delta15N and the first-year N response to fertilizer within the riparian woody plant community. Future papers will describe the transfer of allochthonous litter N to the stream and its subsequent processing by stream biota. Pre-treatment delta15N differed between the two creeks (P=0.0002), possibly due to residual salmon nitrogen in one of the creeks. Pre-treatment delta15N of current-year needles was enriched compared to leaf litter, which was in turn enriched compared to needles aged 4 years and older. We conclude that fractionation due to retranslocation occurs in at least two phases. The first phase, which optimizes allocation of N in younger needle age classes, is distinctly different from the second, which conserves N prior to abscission. The delta15N difference between creeks was eliminated by the fertilization (P=0.42). In the two dominant conifer species, Abies lasiocarpa and Picea engelmannii, most fertilizer N was found in the current-year foliage; little was found in older needles and none was detected in litter (P=0.53). The only N fixing shrub species, Alnus incana, took up only a small amount of fertilizer N [mean percent N derived from fertilizer (%Ndff) 5.0+/-1.6% (SE)]. Far more fertilizer N was taken up by other deciduous shrubs (mean %Ndff=33.9+/-4.5%). Fertilizer N made up 25% (+/-4.2%) of the N in deciduous shrub litter. These results demonstrate the feasibility of light labeling with 15N and the potential influence of riparian plant species composition on stream nutrient dynamics via allochthonous leaf litter inputs. PMID- 15133733 TI - [The Baltic States: old and new partners in Europe]. PMID- 15133734 TI - [The health care system in the republic of Estonia]. PMID- 15133735 TI - [The health care system in the republic of Latvia]. PMID- 15133736 TI - [The health care system in the republic of Lithuania]. PMID- 15133737 TI - [Health telematics/telemedicine in the republic of Estonia]. AB - The people of Estonia, who until their independence had been systematically deprived of all forms of information, have shown a ready acceptance of the use of information technology in all areas of life. This environment and an excellent IT infrastructure have since the beginning of the new century provided favourable conditions for developing various individual projects in telemedicine. At the core of current telemedical applications in Estonia is the BITNET Project (in neurology, general medicine), built up with Swedish cooperation, and three German Estonian projects (in telecardiology and telepathology). These projects are accompanied by studies of their cost-effectiveness. They constitute the basis for the plan that routine telemedical services be taken over by the Estonian health insurance. Differing from the situation in the Scandinavian countries, which have had an effective national telemedical service for over ten years, the special feature of telematrics in Estonia is its international networking with foreign centres of excellence. This has its origin in the aim of Estonian health policy to ensure medical services within its own country in all branches of medicine. PMID- 15133738 TI - [Bilateral German-Estonian projects of the German Federal Ministry of Health and Social Security 1991-2004. Model for cooperation in public health with the states of middle and eastern Europe]. PMID- 15133739 TI - [The Estonian Genome Project in the context of European genome research]. AB - It is the aim of the Estonian Genome Project to establish a database which compiles phenotype and genotype data of a large part of the Estonian population. The Gene Bank will only be used for scientific and public health research. Researchers hope that it will help identify disease genes and prepare the ground for the personalized medicine of the future. Additionally, the project will improve Estonian's international competitiveness in high technology and have a strong educational effect on the population. The legal framework, the Human Genes Research Act, was passed by the Estonian parliament in December 2000. It had been drafted by a group of experts who took into account all available international guidance documents on genetic research. With the pilot project involving three selected regions successfully finished, the main project has now started. PMID- 15133740 TI - [Interventional cardiology in Latvia from 1993 to 2003]. AB - A nationale programme for combatting cardiovascular diseases is being implemented in Letvia. A milestone was reached in 1993 with the establishment of a Letvian Centre for Cardiology. Within 10 years the only cardiac catheterization unit in the country has developed into a centre of excellence. But to meet the cardiological needs for all of Letvia requires at least three additional units for left heart catheterization and one for paediatric a cardiac catheterization. PMID- 15133741 TI - [Pregnancy in patients with Eisenmenger's syndrome. Experiences from Vilnius 1967 2003]. AB - Pregnancy in patients with Eisenmengers syndrome is associated with high maternal and fetal mortality rates and effective contraception or termination of pregnancy is strongly recommended. When these patients decide to begin or continue a pregnancy against advice a coordinated multi-specialist care at a center for congenital heart disease is mandatory. So far, experience with this demanding group of patients is limited. This report reviews the outcome of 30 pregnancies in 10 patients from a group of 35 women with Eisenmengers syndrome who have been treated at the University Hospital in Vilnius, Lithuania, between 1967 and 2003. The most frequent underlying lesion was a ventricular septal defect in 5 patients, their mean age was 26.2 +/- 5,4 years and they were in functional class II or III (9/1) at the beginning of the pregnancy. In this group 13 pregnancies were terminated (43 %), 4 spontaneous abortions (13 %) occurred at less than 21 weeks of pregnancy and one stillbirth at 23 weeks of gestation. Twelve children were born alive, 10 were premature and 2 term deliveries. One maternal death occurred on the third day following an emergency delivery in week 36 and another patient decompensated immediately following delivery, but could be stabilized with intensive care therapy. During long-term follow-up all of these 9 patients were alive at 10/2003 despite a significant deterioration in functional class. These data are comparable to previously published observations in pregnant women with Eisenmengers syndrome. PMID- 15133742 TI - [History of German-Baltic relations in medicine]. AB - Today, the three Baltic countries Estonia, Latvia and Lithuania, have well-known medical faculties with international standing. Their individual histories are briefly outlined. However, relations of the German academic world were closest with the university of Dorpat (today: Tartu). It was re-opened in 1802 by tsar Alexander I in order to keep young Baltic people from studying abroad. The medical faculty was its biggest faculty. The university was Russian, but the official language was German. So many a German professor came to Dorpat and many professors from Dorpat were offered a chair at a German university. The scientific imports connected Dorpat with other centres of West-European science, they brought knowledge and ideas and an exchange of information. The standard was high, and among the teaching staff was a handsome number of medical celebrities, e.g. the anatomist August Rauber and the surgeon Ernst von Bergmann. In Dorpat, Rudolf Buchheim brought a new science, experimental pharmacology, into being, which his pupil and successor, Oswald Schmiedeberg, fully established and propagated all over the world. PMID- 15133743 TI - Highly significant linkage to the SLI1 locus in an expanded sample of individuals affected by specific language impairment. AB - Specific language impairment (SLI) is defined as an unexplained failure to acquire normal language skills despite adequate intelligence and opportunity. We have reported elsewhere a full-genome scan in 98 nuclear families affected by this disorder, with the use of three quantitative traits of language ability (the expressive and receptive tests of the Clinical Evaluation of Language Fundamentals and a test of nonsense word repetition). This screen implicated two quantitative trait loci, one on chromosome 16q (SLI1) and a second on chromosome 19q (SLI2). However, a second independent genome screen performed by another group, with the use of parametric linkage analyses in extended pedigrees, found little evidence for the involvement of either of these regions in SLI. To investigate these loci further, we have collected a second sample, consisting of 86 families (367 individuals, 174 independent sib pairs), all with probands whose language skills are >/=1.5 SD below the mean for their age. Haseman-Elston linkage analysis resulted in a maximum LOD score (MLS) of 2.84 on chromosome 16 and an MLS of 2.31 on chromosome 19, both of which represent significant linkage at the 2% level. Amalgamation of the wave 2 sample with the cohort used for the genome screen generated a total of 184 families (840 individuals, 393 independent sib pairs). Analysis of linkage within this pooled group strengthened the evidence for linkage at SLI1 and yielded a highly significant LOD score (MLS = 7.46, interval empirical P<.0004). Furthermore, linkage at the same locus was also demonstrated to three reading-related measures (basic reading [MLS = 1.49], spelling [MLS = 2.67], and reading comprehension [MLS = 1.99] subtests of the Wechsler Objectives Reading Dimensions). PMID- 15133748 TI - Continuous subcutaneous insulin infusion with short-acting insulin analogues or human regular insulin: efficacy, safety, quality of life, and cost-effectiveness. AB - Portable insulin infusion devices are effective and safe insulin delivery systems for managing diabetes mellitus, especially type 1 diabetes. Rapidly absorbed insulin analogues, such as insulin lispro or insulin aspart, may offer an advantage over regular human insulin for insulin pumps. Several open-label randomised crossover trials demonstrated that continuous subcutaneous insulin infusion (CSII) with insulin lispro provided a better control of postprandial hyperglycaemia and a slightly but significantly lower glycated haemoglobin level, with lower daily insulin requirement and similar or even less hypoglycaemic episodes. A CSII study comparing insulin lispro and insulin aspart demonstrated similar results with the two analogues, and better results than those with regular insulin. Because these analogues have a quicker onset and a shorter duration of action than regular insulin, one might expect an earlier and greater metabolic deterioration in case of CSII interruption, but a more rapid correction of metabolic abnormalities after insulin boluses when reactivating the pump. These expectations were confirmed in randomised protocols comparing the metabolic changes occurring during and after CSII interruption of various durations when the pump infused either insulin lispro or regular insulin. The extra cost resulting from the use of CSII and insulin analogues in diabetes management should be compensated for by better metabolic control and quality of life. In conclusion, CSII delivering fast-acting insulin analogues may be considered as one of the best methods to replace insulin in a physiological manner by mimicking meal and basal insulin requirements, without higher risk of hypoglycaemia or ketoacidosis in well-educated diabetic patients. PMID- 15133749 TI - Hyperinsulinemic hypoglycemia of infancy: the challenge continues. AB - Hypoglycemia due to hyperinsulinemia is the most common cause of persistent hypoglycemia in infants and children. Recent discoveries in the molecular and biochemical regulation of insulin secretion have dramatically increased our understanding of the disorders responsible for syndromes of hyperinsulinemic hypoglycemia. Here, we briefly review the current knowledge of disorders of the K(ATP) channel, activating mutations of glucokinase and glutamate dehydrogenase (GDH) and other disorders that may be associated with specific phenotypes and permit appropriate targeted therapies. Despite these advances, much remains to be learned. We do not understand the mechanisms or defects in many instances, including defective carbohydrate glycosylation syndromes and perinatal hypoxia, both of which may be associated with hyperinsulinemia. Most importantly, preoperative distinction between diffuse and focal lesions cannot be always reliably made even after selective arterial infusion with calcium, glucose or a sulfonylurea with concurrent hepatic venous sampling for insulin. The ability to distinguish diffuse from localized lesions has profound implications for therapeutic approaches, prognosis and genetic counseling. To date, about 50% of individuals with hyperinsulinemic hypoglycemia of infancy can be correctly categorized. Thus, the challenge continues. PMID- 15133750 TI - Skeletal involvement in patients with diabetes mellitus. AB - Studies on skeletal involvement in patients with diabetes mellitus have generated conflicting results, largely because of the pathogenetic complexity of the condition. Several mechanisms may contribute to skeletal damage, including the increased urinary excretion coupled with the lower intestinal absorption of calcium, the inappropriate homeostatic response in terms of parathyroid hormone secretion, and also the complex alteration of vitamin D regulation. Decreased or increased insulin and IGF-1 concentrations and the effects of the accumulation of glycation endproducts on the bone tissue could also play a role. A possible genetic predisposition is also currently under investigation. Finally, the role of fat tissue in type 1 and type 2 diabetes and that of diabetic complications also deserve note. As far as bone mass is concerned, in adult patients with type 1 diabetes a moderately reduced bone mineral density has been shown in both axial and appendicular skeleton. On the contrary, patients with type 2 diabetes seem to have higher bone mineral density in respect to healthy control subjects, especially when overweight women are considered. No clear relationship between bone mass measurements and biochemical parameters of mineral metabolism has been shown in the different types of diabetes. Cohort studies recently carried out on large samples indicate that diabetic patients (with both type 1 and type 2 disease) have a higher risk for fracture, in particular for hip fracture, the most dangerous osteoporotic complication. This seems to be dependent both on qualitative and quantitative alterations of the bone, as well as on extra skeletal factors due to the neuropathic and microangiopathic complications of the disease. PMID- 15133751 TI - CRP and IL-6 concentrations are associated with poor glycemic control despite preserved beta-cell function during the first year after diagnosis of type 1 diabetes. AB - BACKGROUND: The role of non-specific inflammation in beta-cell loss in type 1 diabetes is unclear. In the present study, inflammatory markers were determined in patients with newly diagnosed disease and related to beta-cell function, glycemic control and autoimmunity. METHODS: Ninety-seven adult patients with type 1 diabetes mellitus (80% islet antibody positives, ab(+)) were examined at diagnosis and 3, 6, 9 and 12 months after the start of insulin treatment. Plasma C-reactive protein (CRP), interleukin-6 (IL-6), C-peptide, islet autoantibodies, insulin requirement and HbA(1c) were assessed. RESULTS: The concentrations of CRP were high-normal at diagnosis and did not change during the study period. A positive correlation between CRP at diagnosis and BMI was observed in ab(+) as well as in ab(-) cases. Detectable concentrations of IL-6 were found in 32% (157/485) of the samples and did not change during the study. Ab(-) patients had higher values of CRP at diagnosis and throughout the study compared to the ab(+). Among the ab(+) patients, CRP concentrations during the study were positively correlated to C-peptide at 12 months and an increase in HbA(1c) levels between 6 and 12 months. No associations between the presence or levels of islet autoantibodies and CRP were noted. CONCLUSIONS: In type 1 diabetes, the islet destructive process and the development of beta-cell remission are not associated with changes in CRP or IL-6. Instead, elevated CRP concentrations are prevalent and seem to reflect insulin resistance, as positive associations to BMI, C peptide and deterioration of glycemic control were observed. PMID- 15133752 TI - Amelioration of the beta-cell dysfunction in diabetic APA hamsters by antioxidants and AGE inhibitor treatments. AB - BACKGROUND: In our recent report, probucol treatment ameliorated glucose intolerance and increased the insulin-positive area in the pancreas of streptozotocin (SZ)-induced diabetic APA hamsters. The data suggested that the beneficial effects of probucol treatment on beta-cell function might result from its additive effect as an antioxidant. Here, we examined the antioxidant effects on the beta-cell function in SZ-induced diabetic APA hamsters treated with three different agents, N-acetyl-L-cysteine (NAC), aminoguanidine (AG) and pyridoxamine (PM). METHODS: The control (CB group) and diabetic (SZ group) hamsters were treated with NAC, AG or PM for four weeks from several days after SZ injection. RESULTS: Non-fasting plasma glucose and glycoalbumin levels were significantly reduced in SZ animals by NAC or PM treatment. Glucose tolerance test revealed that fasting plasma glucose levels of SZNAC and SZPM animals were low, similar to the corresponding control animals. Thirty minutes after glucose injection, amelioration in the plasma glucose levels of SZNAC and SZPM animals was observed. Immunohistochemistry revealed that the pancreata of SZNAC, SZAG and SZPM animals showed significantly higher percentages of insulin-positive area than those of non-treated SZ animals. The plasma 8OHdG and malondialdehyde plus 4-hydroxy-2 nonenal (4HNE) levels were significantly decreased especially in SZNAC and SZPM animals. 4HNE-positive cells stained by anti-4HNE antibody were reduced in the islets of each agent-treated animal. SZNAC and SZPM animals showed significantly increased beta-cell proliferation determined by insulin and BrdU double staining. All SZ groups treated with NAC, AG or PM had the high expression levels of Reg and INGAP, which are known to be expressed in regenerating islets. CONCLUSIONS: These data suggest that NAC and PM treatment of SZ-injected diabetic hamsters reduces oxidative stress and restores beta-cell function, but that AG treatment has little beneficial effect on the diabetic conditions of SZ-injected hamsters. PMID- 15133753 TI - A new type 1 diabetes susceptibility locus containing the catalase gene (chromosome 11p13) in a Russian population. AB - BACKGROUND: Oxidative stress is involved in the origin of type 1 diabetes. Low efficiency of the scavenging antioxidant system has been shown to be related to the pathogenesis of the disease. This, therefore suggests that genes encoding catalase and other antioxidant enzymes may implicate in the development of type 1 diabetes. METHODS: Nine microsatellite markers that cover about 10 megabases around the catalase (CAT) gene on chromosome 11p13 were analyzed using polymerase chain reaction (PCR) and fluorescence-based genotyping on an automatic DNA sequencer. We also evaluated three single-nucleotide polymorphisms (SNP) within genes encoding catalase (T1667T and C(-262)T dimorphism) and ETS homologous factor (EHF) (C255T SNP) using a PCR-restriction fragment-length polymorphism approach. Multipont linkage analysis in 37 affected sibling pairs was performed using GENEHUNTER 2.1. We examined the markers for association with the disease by transmission disequilibrium tests in 57 discordant sibling pairs and by a case control study in 258 unrelated healthy donors and 247 affected patients. RESULTS: We obtained close-to-suggestive evidence of linkage to type I diabetes, with the maximum linkage peak between markers D11S907 and D11S2008. Analysis of three SNPs at the CAT and EHF gene located within the region of maximum linkage showed that T1667T and C(-262)T markers of the CAT gene are strongly associated with the disease. CONCLUSION: Our findings support evidence of a new putative type 1 diabetes susceptibility locus on chromosome 11p13 and suggest that the CAT gene may play a role in conferring susceptibility to the disorder in Russian patients. PMID- 15133754 TI - Effects of glipizide GITS and glibenclamide on metabolic control, hepatic glucose production, and insulin secretion in patients with type 2 diabetes. AB - OBJECTIVE: Evaluation of effects of glipizide gastrointestinal therapeutic system (GITS) administered once daily (AM or PM) and glibenclamide on glycemic control, insulin secretory response, and hepatic glucose production (HGP) in patients with type 2 diabetes. METHODS: In a randomized, double-blind, and placebo-controlled study, subjects (HbA(1c) between 8.6 and 10.0%) received a titrated daily dose (5 20 mg) of either glipizide GITS AM (n = 11), glipizide GITS PM (n = 10), glibenclamide (n = 11), or placebo (n = 10) for eight weeks. Fasting and 24-h glucose and insulin, HGP, fructosamine, and HbA(1c) were measured at baseline and at study conclusion; glucose and insulin were also evaluated after Sustacal challenge. RESULTS: Fasting and 24-h glucose were significantly reduced by glipizide GITS AM (33%, p < 0.001; 39%, p < 0.0001), glipizide GITS PM (33%, p < 0.0001; 32%, p < 0.0001), and glibenclamide (37%, p < 0.05; 37%, p < 0.0001). Fasting insulin was not significantly increased by any treatment; 24-h insulin was not increased by glipizide GITS AM, but was elevated by glipizide GITS PM (39%, p < 0.05) and glibenclamide (23%, p < 0.05). Fructosamine and HbA(1c) were significantly reduced by glipizide GITS AM (28%, p < 0.001; 22%, p < 0.0001), glipizide GITS PM (25%, p < 0.005; 24%, p < 0.005), and glibenclamide (17%, p < 0.001; 14%, p < 0.05). Glipizide GITS AM and glibenclamide significantly reduced HGP by approximately 19% (p < 0.05) and 17% (p < 0.01) respectively. Glipizide GITS and glibenclamide significantly (p < 0.0001) decreased the glucose excursion after Sustacal challenge. The reductions in glucose excursions were accompanied by significant (p < 0.05) increases in the insulin response, suggesting an improvement in meal-related insulin secretion. CONCLUSIONS: Glipizide GITS and glibenclamide treatment are effective agents for improving fasting plasma glucose and HbA(1c). Each possessed a suppressive effect on basal HGP and improved postprandial glycemia, but only glipizide GITS AM was effective without causing a persistent elevation in insulin. This profile of glipizide GITS AM is therapeutically attractive, as it is consistent with the potential for a reduced risk of hypoglycemia. PMID- 15133755 TI - The antidiabetic agent, gliclazide, reduces high insulin-enhanced neutrophil transendothelial migration through direct effects on the endothelium. AB - BACKGROUND AND AIM: Many lines of evidence indicate that hyperinsulinemia might be associated with coronary atherosclerosis, and, currently, there are no effective strategies for preventing this. We previously reported that high insulin enhances neutrophil-transendothelial migration, a process that involves increased surface presentation of platelet endothelial cell adhesion molecule-1 (PECAM-1) through a mitogen-activated protein (MAP) kinase-dependent event. In this current study, we examined if antidiabetic agents, especially K(ATP) channel blockers, might similarly protect against the leukocyte-endothelial cell interactions enhanced by high insulin. METHODS: Neutrophils transmigration across umbilical vein endothelial cells (in high insulin medium) with or without K(ATP) channel blockers was performed. Neutrophil migration was quantified by measuring myeloperoxidase, and surface expression of endothelial PECAM-1 was examined using cell-surface enzyme immunoassay. RESULTS: Neutrophil-transendothelial migration and PECAM-1 expression were enhanced by insulin (100 micro U/mL, 24 h) and were attenuated by gliclazide (20 micro M), but not by other K(ATP) channel blockers (glibenclamide, nateglinide, and glimepiride). Neutrophil migration and PECAM-1 expression were also increased by the mitogen-activated protein (MAP) kinase activator, anisomycin (1 micro M), and also attenuated by gliclazide. Nitric oxide (NO) synthase inhibitors did not modify either gliclazide effect. CONCLUSIONS: Our results suggest that the K(ATP) channel blocker, gliclazide, blocks high insulin-mediated neutrophil migration and PECAM-1 expression. These gliclazide effects may be mediated through the inhibition of MAP kinase activation and are unrelated to NO production. PMID- 15133756 TI - Mortality in type 2 diabetic subjects prescribed metformin and sulphonylurea drugs in combination: cohort study. AB - BACKGROUND: In the UK Prospective Diabetes Study, treatment with sulphonylurea and metformin in combination was associated with increased mortality. We compared mortality in subjects treated either with metformin or sulphonylurea drugs alone, or in combination. METHODS: Cohort study in 263 general practices in the United Kingdom. Subjects were aged >/=30 years. Outcome was survival from first prescription of oral hypoglycaemic drugs till death from any cause. Transfer to metformin and sulphonylurea in combination was modelled as a time-dependent covariate. Hazard ratios were adjusted for age, sex, year of treatment, presence of coronary heart disease or prescription of cardiovascular drugs. Analyses were also stratified by propensity score. RESULTS: There were 8488 subjects who were initially prescribed sulphonylureas with a total of 20 783 person years of follow up and 1157 deaths. The crude mortality rate was 58.56 per 1000 person years during suphonlyurea as sole treatment. In 1868 subjects who were prescribed additional metformin, the mortality rate was 39.75 per 1000. The adjusted hazard ratio was 1.06 (95% confidence interval 0.85 to 1.31, P = 0.616). There were 3099 subjects initially treated with metformin with a total of 7306 person years of follow-up and 176 deaths. During metformin-only treatment, the mortality rate was 25.48 per 1000. After addition of sulphonylurea in 867 subjects, mortality was 19.35 per 1000. The adjusted hazard ratio was 0.95 (0.64 to 1.40, P = 0.801). CONCLUSIONS: In this large non-randomized study, there was no evidence of increased mortality risk following prescription of sulphonylurea and metformin in combination, as compared to either drug prescribed singly. PMID- 15133758 TI - Current literature in diabetes. PMID- 15133757 TI - Improved insulin secretory function and reduced chemotactic properties after tissue culture of islets from type 1 diabetic patients. AB - BACKGROUND: Type 1 diabetes results from the destruction of pancreatic beta-cell as a consequence of an autoimmune process. To date, information on the properties of islets isolated from type 1 diabetic patients is very scant. METHODS: Some immunological and functional properties of islets prepared from the pancreas of type 1 diabetic patients were studied shortly after the isolation and after a period of culture in euglycemic condition. RESULTS: Compared to control islets, freshly prepared type 1 diabetic islets released a significantly higher amount of cytokines (pg/mL) into the culture medium (TNF-alpha: 112.9 +/- 5.6 vs 75.6 +/- 24.4; INF-gamma: 286.9 +/- 26.9 vs 58.6 +/- 6.2; IL-10: 41.8 +/- 4.3 vs 10.1 +/- 3.2; TGF-1 beta: 294.0 +/- 20.6 vs 45.1 +/- 3.5); had a significantly higher chemotactic index (1.9 +/- 0.2 vs 1.2 +/- 0.1); showed reduced insulin release (% of insulin content) in response to glucose (2.8 +/- 0.7 vs 5.3 +/- 1.9), arginine (3.0 +/- 0.6 vs 5.6 +/- 1.0), and glibenclamide (2.9 +/- 0.7 vs 5.4 +/- 0.9); and exhibited decreased glucose oxidation capability and diminished mRNA expression of glucokinase, aldolase, pyruvate kinase, and mitochondrial glycerolphosphate dehydrogenase. Ten days after isolation, a normalization of cytokine release (TNF alpha: 55.7 +/- 2.3; INF-gamma: 53.9 +/- 4.3; IL-10: 8.6 +/- 0.7; TGF-1 beta: 60.7 +/- 12.4) and chemotactic index (1.3 +/- 0.2) were observed. Moreover, there was an improvement of insulin secretion (3.8 +/- 0.3, 4.7 +/- 0.6 and 3.5 +/- 0.2 respectively in response to glucose, arginine, and glibenclamide) and glucose oxidation, and a partial recovery of the measured mRNA expressions. CONCLUSIONS: These novel results, obtained with islets prepared from patients with type 1 diabetes, demonstrate that even after months after diabetes diagnosis, a period of culture of the islets in a more favorable environment has beneficial effects on the islet function. PMID- 15133760 TI - Gene therapy for lysosomal storage diseases: the lessons and promise of animal models. AB - There are more than 40 different forms of inherited lysosomal storage diseases (LSDs) known to occur in humans and the aggregate incidence has been estimated to approach 1 in 7000 live births. Most LSDs are associated with high morbidity and mortality and represent a significant burden on patients, their families, and health care providers. Except for symptomatic therapies, many LSDs remain untreatable, and gene therapy is among the only viable treatment options potentially available. Therapies for some LSDs do exist, or are under evaluation, including heterologous bone marrow transplantation (BMT), enzyme replacement therapy (ERT), and substrate reduction therapy (SRT), but these treatment options are associated with significant concerns, including high morbidity and mortality (BMT), limited positive outcomes (BMT), incomplete response to therapy (BMT, ERT, and SRT), life-long therapy (ERT, SRT), and cost (BMT, ERT, SRT). Gene therapy represents a potential alternative therapy, albeit a therapy with its own attendant concerns. Animal models of LSDs play a critical role in evaluating the efficacy and safety of therapy for many of these conditions. Naturally occurring animal homologs of LSDs have been described in the mouse, rat, dog, cat, guinea pig, emu, quail, goat, cattle, sheep, and pig. In this review we discuss those animal models that have been used in gene therapy experiments and those with promise for future evaluations. PMID- 15133761 TI - Portal branch ligation induces efficient retrovirus-mediated gene delivery in rat liver. AB - BACKGROUND: The in vivo transduction of hepatocytes with conventional retrovirus vectors requires the induction of cell division and this can currently only be achieved by invasive surgery or by inducing severe liver damage. We hypothesised that partial portal branch ligation (PBL) could induce hepatocyte proliferation and efficient gene transfer in the rat. METHODS: We ligated the portal branch serving 70% of the liver and measured the kinetics of liver mass restoration and cell proliferation and the distribution of dividing hepatocytes after administration of 5-bromo-2'-deoxyuridine. The efficiency of retrovirus-mediated gene transfer after PBL was tested by use of beta-galactosidase-expressing recombinant retroviruses. The viruses were administered in a single injection via the portal vein at different times after PBL and the livers of transduced animals were analysed 4 days later. RESULTS: We found that the number of cycling hepatocytes remained stable between 24 and 44 h after PBL (approximately 12.5%). Although there was a high level of inter-animal variability, hepatocyte proliferation was always initiated in the same lobe of the liver. In animals that had undergone PBL, 19% of hepatocytes were transduced 28 h after the administration of a single high-titre injection of retroviruses, mainly around the portal spaces. CONCLUSIONS: PBL can mediate the efficient transduction of hepatocytes in vivo after a single intravenous injection of recombinant retroviruses. This approach is feasible in humans. PMID- 15133762 TI - 5-Fluorouracil or gemcitabine combined with adenoviral-mediated reintroduction of p16INK4A greatly enhanced cytotoxicity in Panc-1 pancreatic adenocarcinoma cells. AB - BACKGROUND: Pancreatic cancer is one of the most lethal of all the common gastrointestinal malignancies. Although surgery offers the best chance for survival, it is not appropriate for all cases. The only adjuvant treatment to show promise is chemotherapy. Hence new treatments are urgently sought. We previously reported that adenoviral (Ad)-mediated delivery of p53 (Adp53) and p16(INK4A) (Adp16) significantly inhibited the growth of pancreatic cancer cell lines and established subcutaneous pancreatic tumours in nude mice (Ghaneh P, et al. Adenovirus mediated transfer of p53 and p16INK4A results in pancreatic cancer regression in vitro and in vivo. Gene Ther 2001; 8: 199-208). In this study we examine whether combining Ad-mediated delivery of p53 or p16(INK4A) with clinically relevant chemotherapeutic drugs has therapeutic potential for pancreatic cancer. METHODS AND RESULTS: Four pancreatic adenocarcinoma cell lines were evaluated for their sensitivity to 5-fluorouracil (5-FU) and gemcitabine and two of these, Suit-2 and Panc-1, were chosen for combination experiments because they showed moderate and poor sensitivity, respectively, to 5-FU and gemcitabine. We found no evidence for enhanced cytotoxicity when either cell line was transduced with Adp53 before or after incubation with chemotherapeutic drugs. In contrast, incubation of Panc-1 cells with either 5-FU or gemcitabine followed by Ad-mediated overexpression of p16(INK4A) resulted in a substantial reduction in cell viability under conditions where the drugs alone had minimal cytotoxicity. Incubation of Suit-2 cells with 5-FU followed by Ad-mediated overexpression of p16(INK4A) also resulted in a significant reduction in cell viability. This, however, was observed only with higher concentrations of 5-FU and viral vector. Cell cycle analysis of Panc-1 cells showed that the combination of cytotoxic drugs and Adp16 resulted in an increase in the sub-G1 population suggesting an increase in apoptosis. Dual labelling of these cells with annexin V and propidium iodide (PI) confirmed that the combination of 5-FU and Adp16 resulted in a significant increase in early apoptotic cells (annexin V positive and PI negative) compared with controls. Moreover, overexpression of p16(INK4A) was associated with a reduction in pRb levels in these cells-high levels of pRb have been proposed to contribute to chemoresistance in pancreatic cancer cells. CONCLUSIONS: We have shown that the currently used chemotherapeutic drugs for pancreatic adenocarcinoma combined with restoration of p16(INK4A) expression hold promise for the adjuvant treatment of this disease. Importantly, the combination facilitated the use of chemotherapeutic drugs at lower concentrations than would otherwise be effective. PMID- 15133763 TI - Impact of novel histone deacetylase inhibitors, CHAP31 and FR901228 (FK228), on adenovirus-mediated transgene expression. AB - BACKGROUND: Histone deacetylase inhibitors (HDIs) are known to enhance adenovirus (Ad)-mediated transgene expression. Recently, novel HDIs, including cyclic hydroxamic-acid-containing peptide 31 (CHAP31) and FR901228 (FK228), have been developed. METHODS: The effects of these two novel HDIs on Ad-transduced or endogenous gene expression were investigated. Acetylation of core histones and the expression of the coxsackie and adenovirus receptor (CAR) in HDI-treated cells were examined using Western blot and a quantitative reverse transcription polymerase chain reaction (TaqMan RT-PCR), respectively. Their in vivo effect on adenoviral gene expression was investigated in BALB/c mice. RESULTS: Both compounds enhanced and prolonged Ad-mediated beta-galactosidase expression more effectively than did trichostatin A, a classic HDI. The same effect was observed in Ad-transduced heat shock protein 72 (HSP72), but not in hyperthermia-induced endogenous expression of HSP72, suggesting that the effect is specific for transduced gene expression. Hyperacetylation of core histones induced by HDIs was considered responsible for the augmentative effects of gene expression. Intravenous administration of either CHAP31 or FR901228 enhanced beta galactosidase expression in mice infected with AdLacZ. CONCLUSIONS: CHAP31 and FR901228 amplified Ad-mediated transgene expression. The enhancement of transgene expression by HDIs may result in fewer vector doses for necessary gene expression, helping to alleviate disadvantages caused by Ad vectors. This could be a useful tool in overcoming current limitations of gene therapy using adenovirus vectors. PMID- 15133764 TI - Comparative analysis of dendritic cells transduced with different anti-apoptotic molecules: sensitivity to tumor-induced apoptosis. AB - BACKGROUND: Tumors develop mechanisms to escape recognition by the immune system. It has recently been demonstrated that tumors cause apoptotic death of key immune cells, including the major antigen-presenting cells, dendritic cells (DC). Elimination of DC from the tumor environment significantly diminishes development of specific immunologic responses. We have recently demonstrated that tumor induced DC apoptosis could be prevented by overexpression of the anti-apoptotic molecule Bcl-x(L). The aim of this study was to identify extrinsic and intrinsic tumor-induced apoptotic pathways in DC by targeting different anti-apoptotic molecules, including FLIP, XIAP/hILP, dominant-negative procaspase-9 and HSP70. METHODS: Murine bone marrow derived DC were transduced with adenoviral vectors carrying different anti-apoptotic molecules and co-incubated with tumor cells in a Transwell system. Apoptosis of DC was assessed by Annexin V and PI staining. RESULTS: We have demonstrated that adenoviral infection of DC with genes encoding different anti-apoptotic molecules exhibits different degrees of resistance to melanoma-induced apoptosis. Furthermore, we have shown that anti-apoptotic molecules other than the Bcl-2 family of proteins are able to protect DC and prevent tumor-induced apoptosis in DC. CONCLUSIONS: The results show that tumor induced apoptosis of DC is not limited to the mitochondrial pathway of cell death and open additional possibilities for targeted molecular protection of DC longevity in cancer. Therefore, effective protection of DC from tumor-induced apoptosis may significantly improve the efficacy of DC-based therapies for cancer. PMID- 15133765 TI - Gene transfer into rabbit arteries with adeno-associated virus and adenovirus vectors. AB - BACKGROUND: Gene transfer offers considerable potential for altering vessel wall physiology and intervention in vascular disease. Therefore, there is great interest in developing optimal strategies and vectors for efficient, targeted gene delivery into a vessel wall. METHODS: We studied adeno-associated viruses (AAV; 9 x 10(8) to 4 x 10(9) TU/ml) for their usefulness to transduce rabbit arteries in vivo in comparison with adenoviruses (Adv; 1 x 10(9) to 1 x 10(10) pfu/ml). 100 microl of viruses or placebo solution were injected intraluminally into transiently isolated carotid segments. RESULTS: In normal arteries AAV transduced mainly medial smooth muscle cells (SMC) while Adv transduced exclusively endothelial cells (EC). Mechanical injury to EC layer and internal elastic lamina enabled Adv to penetrate and transduce medial SMC. Transgene expression in EC after the AAV-mediated gene transfer was very low. The use of the EC-specific Tie-1 promoter did not lead to specific transgene expression in EC. Transgene expression in SMC persisted for at least 100 days after the AAV treatment whereas the Adv-mediated effect diminished in 14 days. AAV caused only a modest increase in EC VCAM-1 expression and proliferation rate of vascular cells as compared with the mock-treated arteries while Adv caused an extensive inflammatory cell infiltration, VCAM-1 expression, vascular cell proliferation and morphological damages. CONCLUSIONS: Significant differences were observed between the AAV and the Adv vectors in their patterns of arterial transduction and consequent inflammatory responses. These distinct properties may be utilized for different applications in vascular biology research and gene therapy for cardiovascular diseases. PMID- 15133766 TI - Identification of a replication-defective herpes simplex virus for recombinant adeno-associated virus type 2 (rAAV2) particle assembly using stable producer cell lines. AB - BACKGROUND: The development of stable producer cell lines for recombinant adeno associated virus (rAAV) assembly is a strategy followed by many groups to develop scalable production methods suitable for good manufacturing practice (GMP) requirements. The major drawback of this method lies in the requirement for replicating adenovirus (Ad) for rAAV assembly. In the present study, we analyzed the ability of several replication-defective herpes simplex type 1 (HSV-1) helper viruses to induce rAAV2 particle production from stable producer cell lines. METHODS: Several stable rAAV producer cell clones were infected with wild-type and replication-defective HSV strains and analyzed for rep-cap gene amplification, viral protein synthesis and rAAV titers achieved. In vivo analysis following rAAV injection in the murine brain was also conducted to evaluate the toxicity and biopotency of the rAAV stocks. RESULTS: We demonstrated that an HSV strain mutated in the UL30 polymerase gene could efficiently be used in this context, resulting in rAAV titers similar to those measured with wild-type HSV or Ad. Importantly, with respect to clinical developments, the use of this mutant resulted in rAAV stocks which were consistently devoid of contaminating HSV particles and fully active in vivo in the murine central nervous system with no detectable toxicity. CONCLUSIONS: This study, together with our previous report describing a rAAV chromatography-based purification process, contributes to the definition of an entirely scalable process for the generation of rAAV particles. PMID- 15133767 TI - The role of polyadenylation signal secondary structures on the resistance of plasmid vectors to nucleases. AB - BACKGROUND: Nuclease degradation of plasmid DNA (pDNA) vectors after delivery and during trafficking to the nucleus is a barrier to gene expression. This barrier may be circumvented by shielding the pDNA from the nuclease-rich cell environment with adjuvants or by using nuclease inhibitors. A different alternative that is explored in this work is to make pDNA vectors more nuclease-resistant a priori. METHODS AND RESULTS: The hypothesis that a significant part of nuclease attack is directed towards certain labile sequences in a pDNA model (pVAX1/lacZ) was first tested. Homopurine-rich tracts in the bovine growth hormone polyadenylation signal (BGH poly A) were identified as labile sequences using S1 nuclease as a probe. Two pDNA variants were then created by replacing the BGH poly A region with the SV40 or a synthetic poly A signal. A study of plasmid degradation in eukaryotic cell lysates and mice plasma showed that the half-life of the supercoiled isoforms of the new vectors was always higher when compared with the control plasmid. An in vitro assay of the reporter beta-galactosidase in transfected CHO cells further showed that gene expression with the new pDNA variants was not affected negatively by the plasmid modifications. CONCLUSIONS: The replacement of labile sequences in plasmid DNA vectors improves resistance towards nuclease attack as shown by the increased half-lives of supercoiled plasmid isoforms incubated with endo/lysosomal, cytoplasmatic and blood plasma enzymes. PMID- 15133768 TI - Excision of Sleeping Beauty transposons: parameters and applications to gene therapy. AB - A major problem in gene therapy is the determination of the rates at which gene transfer has occurred. Our work has focused on applications of the Sleeping Beauty (SB) transposon system as a non-viral vector for gene therapy. Excision of a transposon from a donor molecule and its integration into a cellular chromosome are catalyzed by SB transposase. In this study, we used a plasmid-based excision assay to study the excision step of transposition. We used the excision assay to evaluate the importance of various sequences that border the sites of excision inside and outside the transposon in order to determine the most active sequences for transposition from a donor plasmid. These findings together with our previous results in transposase binding to the terminal repeats suggest that the sequences in the transposon-junction of SB are involved in steps subsequent to DNA binding but before excision, and that they may have a role in transposase-transposon interaction. We found that SB transposons leave characteristically different footprints at excision sites in different cell types, suggesting that alternative repair machineries operate in concert with transposition. Most importantly, we found that the rates of excision correlate with the rates of transposition. We used this finding to assess transposition in livers of mice that were injected with the SB transposon and transposase. The excision assay appears to be a relatively quick and easy method to optimize protocols for delivery of genes in SB transposons to mammalian chromosomes in living animals. PMID- 15133769 TI - Hydrodynamics-based procedure involves transient hyperpermeability in the hepatic cellular membrane: implication of a nonspecific process in efficient intracellular gene delivery. AB - BACKGROUND: The mechanisms underlying the efficient gene transfer by a large volume and high-speed intravenous injection of naked plasmid DNA (pDNA), a so called hydrodynamics-based procedure, remain unclear and require further investigation. In this report, we have investigated possible mechanisms for the intracellular transport of naked pDNA by this procedure. METHODS: Propidium iodide (PI), a fluorescent indicator for cell membrane integrity, and luciferase- or green fluorescent protein (GFP)-expressing pDNA were injected into mice by the hydrodynamics-based procedure. RESULTS: PI was efficiently taken up by hepatocytes which appeared to be viable following the hydrodynamics-based procedure. Pre-expressed GFP in the cytosol was rapidly eliminated from the hepatocytes by a large-volume injection of saline. The profiles of plasma ALT and AST showed a steady decline with the highest values observed immediately after the hydrodynamics-based procedure. These results suggest that the hydrodynamics based procedure produces a transient increase in the permeability of the cell membrane. The cellular uptake process appeared nonspecific, since simultaneous injection of an excess of empty vector did not affect the transgene expression. Sequential injections of a large volume of pDNA-free saline followed by naked pDNA in a normal volume revealed that the increase in membrane permeability was transient, with a return to normal conditions within 30 min. Transgene expression was observed in hepatocyte cultures isolated 10 min after pDNA delivery and in the liver as early as 10 min after luciferase-expressing RNA delivery, indicating that pDNA delivered immediately by the hydrodynamics-based procedure has the potential to produce successful transgene expression. CONCLUSIONS: These findings suggest 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 increased permeability. PMID- 15133771 TI - Psychosocial problems among younger women with breast cancer. AB - Women diagnosed with breast cancer at a younger age often have concerns less frequently faced by older women. A cross-sectional survey of 204 women diagnosed with breast cancer at age 50 or younger within the past 3.5 years was conducted to examine issues particularly faced by younger women. The questionnaire included standardized measures of problems related to breast cancer (CARES) and open-ended questions asking women about their experiences. Concerns about premature menopause and pregnancy related issues, among those women for whom these issues were applicable, had the highest ratings as problems experienced by women since their diagnosis. Among all women, sexual functioning was a greater problem than lack of sexual interest, and body image was of moderate concern. Overall, relationships with partners were not a problem. In multivariate analyses, having a mastectomy was associated with greater problems with body image and interest in sex. Chemotherapy was associated with greater sexual dysfunction. Responses to open-ended questions were particularly informative and reflected the diversity of responses women have to breast cancer. Findings reinforce the need to develop interventions to help women deal with premature menopause and problems with sexual functioning following chemotherapy. PMID- 15133772 TI - Pessimism as a predictor of emotional morbidity one year following breast cancer surgery. AB - The prevalence of and predictive factors for emotional morbidity (measured by the Hospital Anxiety and Depression Scale (HAD)) one year following surgery, with special focus on dispositional optimism/pessimism (measured by the Life Orientation Test (LOT-R), was examined in 165 women, newly diagnosed with breast cancer. Patients characteristics, appraisal of cancer diagnosis, beliefs about treatment efficacy, treatment decision-making participation, coping and emotional morbidity was assessed by self-rating questionnaires. Prevalence of anxiety and depression cases at time of diagnosis was 34 and 12%, respectively, and 26 and 9% after one year. Prevalence of emotional morbidity was significantly enlarged among pessimists at all assessments. Pessimism was the strongest predictor for anxiety (OR: 0.86 C.I. 95% 0.77 - 0.95) and depression (OR: 0.83, C.I. 95% 0.73 - 0.95) one year following breast cancer surgery. Optimists and pessimists differed not only in regard to coping styles, but also in regards to predictors of emotional morbidity. Optimists experiencing anxiety at time of breast cancer diagnosis had about six times higher risk of experiencing anxiety after one year, compared to optimists without preoperative anxiety. For pessimists, the more pessimistic one was about one's overall future the higher risk for developing anxiety following one year of breast cancer surgery. Pessimists, who endorse helpless/hopeless coping style when receiving a diagnosis of breast cancer, had three times greater risk for experiencing depression one year after breast cancer surgery, than pessimists who did not. Health care professionals should therefore provide intervention for pessimists, as well as for patients with high anxiety scores at time of diagnosis. PMID- 15133773 TI - Effects of a brief intervention program for patients with cancer and their partners on feelings of inequity, relationship quality and psychological distress. AB - When one member of a couple develops a serious illness, the lives of both partners are likely to be affected. Interventions directed at both partners are generally lacking, however. In the present study, a brief counseling program directed at couples confronted with cancer was evaluated. The intervention focused mainly on the exchange of social support and help between both partners and was aimed at restoring perceptions of equity. Couples were randomly assigned to an experimental group or a waiting-list group. After the intervention, both patients and their partners reported lower levels of perceptions of underinvestment and overbenefit, and higher levels of relationship quality. Moreover, among patients psychological distress decreased after the intervention. These effects were generally maintained until follow-up three months later. Associations between perceptions of equity and relationship quality and psychological distress were also examined. PMID- 15133774 TI - We are talking, but are they listening? Communication patterns in families with a history of breast/ovarian cancer (HBOC). AB - This study explores how family communication patterns and family scripts influence the dissemination of genetic information and the sharing of feelings about genetic inheritance in families of healthy women who have attended a cancer genetics risk clinic because of their family history of breast and, or ovarian cancer. Family scripts are sets of expectations, beliefs, and norms that assign meaning to patterns of interaction, connect generations and provide guidance for action. We conducted an exploratory, qualitative study at a major clinical and research cancer center in the United Kingdom from January through June 2000 approved by the hospital clinical research and ethics committees. Twenty-one semi structured, in-depth interviews were conducted using a purposive sample of women coming to the cancer genetics risk clinic for the first time, supplemented by 5 months of participant observation. We identified several communication patterns: open and supportive; directly blocked, indirectly blocked, self-censored and use of third parties. Some family members shared their feelings and discussed ways of trying to avoid developing breast or ovarian cancer; for others disseminating information or just talking about inherited susceptibility for breast and, or ovarian cancer fell into the script violation category; still others tried to renegotiate their family scripts. PMID- 15133775 TI - Characteristics of cancer patients entering a smoking cessation program and correlates of quit motivation: implications for the development of tobacco control programs for cancer patients. AB - Even though survival and quality of life are adversely affected by smoking among cancer patients, about one-third of cancer patients who smoked prior to their diagnosis continue to smoke. One barrier to the provision of smoking cessation treatments to cancer patients is the paucity of data on the characteristics of cancer patients who continue to smoke and a lack of data on correlates of quit motivation in this population. This descriptive study assessed demographic, medical, smoking history, and psychological characteristics of cancer patients in a smoking cessation program (N=111) and examined these characteristics as correlates of quit motivation. Methods used by patients to quit smoking were also queried. We found that: (1) most patients are Caucasian, married, diagnosed with head and neck (versus lung) cancer, highly addicted to nicotine, and in the contemplation or preparation stage of change; (2) most patients attempt to quit smoking without formal treatment, although 33-50% have used the transdermal nicotine patch, nicotine gum, or bupropion; (3) depressive symptoms, low quitting self-efficacy, low perceived risk, and low perceived benefits of quitting are prevalent among patients, but most patients do not endorse the perceived disadvantages of quitting or fatalistic beliefs; and (4) quit motivation is associated with higher quitting self-efficacy, risk perceptions, and perceived benefits of quitting, lower tobacco use and nicotine addiction, and shorter time since diagnosis. These findings can help guide the development of smoking cessation interventions for cancer patients. PMID- 15133776 TI - Prognostic value of depressive coping and depression in survival of lung cancer patients. AB - The aim of this investigation was to determine whether depressive coping and depression predict shorter survival among lung cancer patients. We conducted a prospective study using an inception cohort with a 3-5-year follow-up. The sample consisted of n = 59 (of n = 69 invited to participate) patients (mean age 65 years, S.D. = 9.7; 81% male) newly diagnosed with small cell lung cancer or non small cell lung cancer Stage III or IV who were scheduled for later chemotherapy and/or radiotherapy at a tertiary care centre. Patients were investigated after their diagnosis and before the beginning of treatment. Depressive coping and depression were assessed using standardized self-report questionnaires (Freiburg Questionnaire of Coping with Illness; Hospital Anxiety and Depression Scale). Depressive coping was associated with shorter survival (hazard ratio 1.75, 95% confidence interval 1.04-2.93, p = 0.034) after adjusting for age, sex, stage, histological classification, and Karnofsky performance status but not treatment type, using the Cox proportional hazards regression. Depression, however, was not linked with survival (hazard ratio 1.05, 95% confidence interval 0.98-1.13, p = 0.18). To conclude, the prognostic value of depressive coping was partially confirmed, warranting further examination of the robustness of this relationship. PMID- 15133777 TI - Benefit the patient, manage the risk: a system goal. PMID- 15133778 TI - Drug switching patterns among patients with rheumatoid arthritis and osteoarthritis using COX-2 specific inhibitors and non-specific NSAIDs. AB - PURPOSE: To compare RA and OA patients' time-to-switch after newly initiating treatment with three most commonly used non-specific (NS)-NSAIDs and two COX-2 inhibitors, celecoxib and rofecoxib. METHODS: Managed care enrollees newly prescribed celecoxib, rofecoxib, ibuprofen, naproxen or diclofenac were identified. Time to switch to a different NS-NSAID or COX-2 specific inhibitor was determined using time-to-event analysis and Cox proportional hazards models were used to estimate the odds ratio (OR) after controlling for potential confounders. RESULTS: The time to 25% of the cohort switching was longer for rofecoxib and celecoxib (159 and 205 days respectively) compared to the three NS NSAIDs (49-78 days). Patients were at the highest risk of switching within the first 100 days of therapy. After adjusting for potential confounding factors, the OR for switching to another NS-NSAID or COX-2 specific inhibitor ranged from 1.74 to 2.35 for the three NS-NSAIDs compared to celecoxib (all comparisons, p < 0.01). Similar findings were obtained when comparing rofecoxib to each of the three NS-NSAIDS (all comparisons, p < 0.01). When COX-2 inhibitors combined were compared to NS-NSAIDS combined, the OR for switching was 1.53 (95% confidence interval = 1.42-1.65; p < 0.01) after adjusting for potential confounders. CONCLUSIONS: Patients on the COX-2 specific inhibitors (celecoxib and rofecoxib) were significantly less likely to switch their therapy than patients on NS-NSAIDS (ibuprofen, naproxen and diclofenac). These results suggest that COX-2 specific inhibitors may be a more effective treatment option when compared with NS-NSAIDs in usual clinical practice. PMID- 15133779 TI - Oral anticoagulants and the risk of osteoporotic fractures among elderly. AB - PURPOSE: Coumadin-based oral anticoagulants are associated with a decrease in bone mass density, but their role in fracture risk is equivocal. Because the use of oral anticoagulants is prevalent among the elderly, as is the risk and morbidity of osteoporotic fractures, the association between osteoporotic fractures and oral anticoagulants needs to be clarified. METHOD: We conducted a case-control study on a 10% random sample of subjects aged 70 years and older enrolled in the Quebec universal health insurance plan between 1992 and 1994. Incident cases of a first osteoporotic fracture were identified by International Classification of Diseases, Ninth Revision codes. Exposure was defined as one or more prescriptions of oral anticoagulants dispensed before the osteoporotic fracture. Ten controls for each case, matched by age and date of osteoporotic fracture, were identified. RESULTS: Among 1523 cases, 48 (3.2%) were ever exposed to oral anticoagulants; among 15,205 controls, 461 (3.0%) were ever exposed (crude odds ratio: 1.0: 95% confidence interval: 0.7-1.5). These negative results persisted after adjusting for potential confounding variables and stratifying exposure into cumulative dose and treatment duration. CONCLUSIONS: Coumadin-based oral anticoagulants are not significantly associated with osteoporotic fractures among the elderly, providing reassurance for elderly patients on long-term oral anticoagulants. PMID- 15133780 TI - A potential bias in safety evaluation during open-label extensions of randomized clinical trials. AB - PURPOSE: To describe a bias that can occur in the analysis of data from certain randomized trials. METHODS AND RESULTS: Although randomized trials are effective at preventing confounding, a potentially strong confounding can arise in certain therapeutic drug trials in which follow-up is extended in an open-label phase that allows switching treatments. Many trials implement eligibility screening that excludes those at high risk of morbidity and mortality. In these trials, disease rates and death rates for the study population can rise rapidly during follow-up as the effect of screening wanes. During the open-label follow-up, a preponderance of patients may switch to the new therapy. If so, then any evaluation of the new therapy that includes follow-up from the open-label phase, as is often the case for safety evaluations, will be confounded. The confounding arises because the person-time experience of those on the new treatment will be more heavily weighted with the open-label phase experience, during which morbidity and mortality rates may be much greater than in the initial phase of follow-up. This confounding may be strong and will be in the direction of making the new treatment look worse, provided that the net switching is toward the new treatment during the open-label phase. CONCLUSIONS: The confounding described here is not prevented by randomization because it develops in a non-randomized add-on analysis to the trial. The bias can be removed, however, by controlling for time since randomization in the analysis of the data. PMID- 15133781 TI - Early onset of hyperkalemia in patients treated with low molecular weight heparin: a prospective study. AB - OBJECTIVE: Data on low molecular weight heparin (LMWH) and induction of hyperkalemia is scarce. Therefore, we conducted a prospective study evaluating potassium levels before and after three days of treatment with the LMWH Enoxaparin. METHODS: Patients treated with the LMWH Enoxaparin in the standard therapeutic dosages were included. Levels of potassium, sodium, urea, creatinine, AST, ALT bicarbonate, pH and platelet counts were examined before and after three days of LMWH treatment. Plasma renin activity (PRA) and aldosterone levels were examined in a subgroup of patients. Potassium levels were correlated with disease states and medications known to affect potassium homeostasis. RESULTS: Ninety seven consecutive patients were enrolled in the study, however, 12 patients were excluded from analysis; therefore 85 patients comprised the study group. The most common reason for exclusion (in 9 patients) was the absence of a second potassium result. We found an increase in potassium levels from 4.26 +/- 0.04 mmol/L at baseline to 4.43 +/- 0.04 mmol/L on the third day of treatment (mean +/- SE), with potassium levels exceeding 5.0 mmol/L in 9% of treated patients. There was no life threatening or symptomatic hyperkalemia. PRA and aldosterone levels did not change significantly during the treatment period. There was no correlation between the increase in potassium levels and diabetes mellitus or treatment with angiotensin converting enzymes inhibitors, angiotensin receptor blockers, beta blockers and non-potassium sparing diuretics. CONCLUSIONS: Potassium levels increase on the third day of treatment with the LMWH Enoxaparin. This effect may be aldosterone independent. PMID- 15133782 TI - Prevalence and determinants of antibiotic resistance in faecal Escherichia coli among unselected patients attending general practitioners in Southwest Germany. AB - PURPOSE: Widespread use of antibiotics is thought to be the main reason for the world-wide increase in antibiotic resistance. Although a great majority of antibiotics are prescribed outside hospitals, little is known about the prevalence and determinants of antibiotic resistance in the general population. METHODS: Escherichia coli (E. coli) was cultured from and minimal inhibitory concentrations against six commonly prescribed antibiotic substances were tested in 750 stool samples of 484 unselected, consecutive outpatients aged 40-74 years attending general practitioners. Odds ratios (OR) and their 95% confidence intervals (CI) for the association between potential risk factors and the prevalence of antibiotic resistance were estimated using generalised estimating equations. RESULTS: Prevalence of E. coli resistance against ampicillin, doxycycline, cotrimoxazole or quinolones was 24%. Current antibiotic use was strongly associated with antibiotic resistance, adjusted OR: 11.1, 95% CI: 2.3 53, but antibiotic resistance was unaffected by antibiotic use stopped weeks before. Recent hospitalisations were the only other significant predictor of an increased prevalence of resistance. CONCLUSIONS: The strong association between current use of antibiotics and colonisation with antibiotic resistant E. coli suggests a major role for selection of resistant strains while using antibiotics that seem to be quickly reversible, though. PMID- 15133783 TI - Do therapeutic indications of antidepressants change from one year to another? AB - PURPOSE: To analyse and describe the pattern of prescriptions for antidepressants by all psychiatrists working for the Canary Islands Health Service (CIHS) during the 4-year period from 1999 to 2002. METHODS: All prescriptions for antidepressants by the ten psychiatrists who worked continuously for the CIHS during the period 1999-2002 in the island of Tenerife (700,000 inhabitants) were collected from the CIHS central database. Global prescription of antidepressant medication for the entire region (1.8 million inhabitants) by any physician working for the public sector was counted and converted into defined daily doses (DDDs). RESULTS: The intensity of prescribing antidepressants increased from 22.1 DDD/1000 inhabitants/day in 1999 to 29.1 DDD/1000 inhabitants/day in 2002, with the five top selective serotonine reuptake inhibitors (SSRIs) being responsible for 83.6% of all the antidepressant prescriptions in the year 2002. A wide variation in individual prescription pattern was evident both between and within each psychiatrist during these years. Working in the same conditions, and with a similar morbidity pattern, one psychiatrist prescribed up to 32,000 DDDs in one year, whereas another colleague only prescribed 600 DDDs in the same period and to the same covered population. The amount of individual variation in prescription pattern highly correlated with the intensity of drug prescribing. CONCLUSIONS: The high inter- and intra-individual variation in antidepressant prescribing could not be linked with personal, structural or morbidity patterns, and the heavy influence of pharmaceutical industry could not be ruled out. PMID- 15133784 TI - Benzodiazepines utilization and self-medication as correlates of stress in the population of Serbia. AB - BACKGROUND: High consumption of benzodiazepines (BDZ) occurs in populations exposed to stress. In the last decade of the 20th century, when the population of Serbia experienced increasing economic hardships due to the civil war in former Yugoslavia, UN sanctions and air raids in 1999, diazepam became the most frequently prescribed drug. This period was also characterized by the free marketing of all drugs, which made them available without prescription. OBJECTIVE: To investigate the consumption and the pattern of use of BDZ in the population of Belgrade and Serbia in the period of 1990-2001. MATERIALS AND METHODS: Data on benzodiazepines prescribing and on wholesale in general population of Belgrade and Serbia were collected. In a cross-sectional study of drug prescribing in general practice data were obtained from 1800 patient records in the primary health care centers in Serbia. Statistical analysis was performed by using standard non-parametric tests. RESULTS: Annual rates of BDZ prescribing in Belgrade from 1990 to 1999 were rather uniform (approx. 25 DDD/1000 inhabitants/day), with slight tendency to decrease. In Serbia as a whole, there were significant differences in the annual prescribing rates over the period 1998 2000. The wholesale of BDZ in Serbia significantly increased between 1991 and 2001, with the peak of 133 DDD/1000 inhabitants/day in 1999. The wholesale of BDZ was significantly greater that the rates of prescribing in corresponding years. Over the 10 year period, the numbers of visits to GPs and psychiatrists and the number of neurotic diagnoses were significantly reduced. The use of BDZ in psychiatric hospital increased significantly in 1999 as compared to 1998, although the number of admissions and the occupancy of hospital beds were reduced. In primary health care, diazepam was the most frequently prescribed drug predominantly for non-psychiatric diagnoses. CONCLUSIONS: It is concluded that in the last decade, the utilization of BDZ was increased in the population of Belgrade and Serbia, indicating a clear trend to self-medication, particularly in the period of acute war crisis. PMID- 15133785 TI - Banning antimicrobial growth promoters in feedstuffs does not result in increased therapeutic use of antibiotics in medicated feed in pig farming. AB - OBJECTIVE: We analysed prescription patterns for medicated feedstuffs to find out whether the ban on nutritive antimicrobial growth promotion introduced in Switzerland in 1999 had caused an increase in the therapeutic use of antimicrobial agents given orally to piglets and fattening pigs. METHODS: From 1996 to 2001, a total of 6427 prescriptions were evaluated for medicated pig feed delivered to pig farms in the Swiss canton of St Gall. Prescribed daily doses (PDD) were derived for 14 active ingredients. The overall amount and the potency of antimicrobial agents were measured in relation to the size of the pig population (PDD/population). RESULTS: The use of antimicrobial agents decreased between 1996 (1200 kg) and 1999 (708 kg) and increased thereafter from 779 kg in 2000 to 936 kg in 2001. The PDD/population (6.1 in 1996 and 3.6 in 1999) remained low (3.3 in 2000 and 3.4 in 2001). The difference between the two parameters can be explained by changes in prescribing patterns, namely a reduction in antimicrobial therapy of respiratory diseases in fattening pigs and a shift to antimicrobial treatment of gastrointestinal-tract infections in piglets using drugs with a high PDD. PMID- 15133786 TI - Factors influencing the spontaneous reporting of adverse drug reactions--the experience of the Slovak Republic. AB - The system of adverse drug reaction (ADR) monitoring in the Slovak Republic is greatly influenced by changes taking place in the current health care delivery system. Under-reporting and reporting biases due to selective ADR reporting provide very serious problems in pharmacovigilance. In the year 2001, the number of reported ADRs increased due to reports of angiotensin converting enzyme (ACE) inhibitor-induced cough; this coincided with limits being imposed on the use of angiotensin-II receptor antagonists. These factors provide an example of selective ADR reporting, deforming the quality of drug safety monitoring. They also indicate that administrative strategies can significantly affect spontaneous reporting activities. PMID- 15133787 TI - A functional definition of prolonged pregnancy based on daily fetal and neonatal mortality rates. PMID- 15133788 TI - Ultrasound in acute urinary retention and retroverted gravid uterus. PMID- 15133789 TI - Sonographic demonstration of brain injury in fetuses with severe red blood cell alloimmunization undergoing intrauterine transfusions. AB - OBJECTIVE: To assess sonographically brain anatomy in fetuses with severe anemia due to red blood cell alloimmunization undergoing intrauterine intravascular transfusions. METHODS: Multiplanar neurosonography was performed in seven consecutive hydropic fetuses undergoing intrauterine transfusions (mean gestational age 22 +/- 2.5 weeks; mean hemoglobin concentration at the first transfusion 2.3 +/- 1.0 g/dL). RESULTS: Abnormal cerebral findings were identified in four out of seven fetuses. An intracerebellar hemorrhage developed in two fetuses after the first transfusion and one fetus that had severe brain edema before the first transfusion was later found to have cystic periventricular leukomalacia. In one fetus unilateral ventriculomegaly was noted after the first transfusion. Two fetuses were terminated. The remaining pregnancies had an uneventful course, the infants were delivered between 34 and 36 gestational weeks and were alive and well at the time of writing. Prenatal diagnosis of brain injury was always confirmed except for the case with ventriculomegaly that underwent spontaneous intrauterine resolution. CONCLUSIONS: Fetuses with extreme anemia due to red blood cell alloimmunization can be salvaged by intrauterine transfusion. In some of these cases brain injury may occur prenatally, and the risk seems to be particularly high when the hemoglobin concentration at the time of the first transfusion is 15 IU/mL at 19-38 weeks' gestation and within 10 days of measurement of fetal hemoglobin concentration in blood obtained either by cordocentesis (n = 43) or at delivery (n = 15). In the RBC isoimmunized pregnancies each of the measured MCA-PSV and hemoglobin concentrations was expressed as a delta value (difference in SDs from the normal mean for gestation). Regression analysis was used to determine the significance of the association between delta MCA-PSV and delta fetal hemoglobin concentration. RESULTS: In the normal pregnancies there was a significant increase in fetal MCA PSV with gestation (mean MCA-PSV = 10(0.0223 x GA + 0.963)). In RBC isoimmunized pregnancies the fetal MCA-PSV was increased and there was a significant association between delta MCA-PSV and delta hemoglobin concentration (delta hemoglobin = (delta MCA-PSV + 0.093)/-0.356; R(2) = 0.638, P < 0.0001). An MCA PSV of mean + 1.5 SDs detected 96% of severely anemic fetuses, with a hemoglobin deficit of at least 6 SDs, for a false-positive rate of 14%. CONCLUSION: Measurement of fetal MCA-PSV is a useful method of assessing fetal anemia. In the clinical management of isoimmunized pregnancies a cut-off in MCA-PSV of mean + 1.5 SDs can identify nearly all severely anemic fetuses with a low false-positive rate. PMID- 15133791 TI - Predicting the severity of fetal anemia using time-domain measurement of volume flow in the fetal aorta. AB - OBJECTIVES: To assess the value of fetal aortic time-domain measurement of volume flow (using color velocity imaging quantification (CVI-Q)) in predicting the severity of fetal anemia. METHODS: This was a prospective observational study, in which 24 pregnant women with suspected fetal anemia due to rising anti-red blood cell antibody titers underwent cordocentesis. The fetal aortic time-domain volume flow was measured before fetal blood sampling for fetal hemoglobin investigation. We examined the correlation between increased fetal aortic time-domain volume flow (>2 SD for gestational age) and fetal anemia (hemoglobin level <2 SD for gestational age). RESULTS: Seventeen fetuses had anemia, and seven had normal hemoglobin. There was a strong correlation between the increase in fetal aortic time-domain volume flow and the drop in hemoglobin value (r = 0.81; P < 0.01). The sensitivity of this technique to predict fetal anemia was 81.3% and the specificity was 71.4%. The mean increase over time in aortic CVI-Q in anemic fetuses was 323.2 mL/min (95% CI, 200.1 to 446.4) compared with 86.9 mL/min (95% CI, -17.7 to 191.5) in the non-anemic group (P = 0.004). CONCLUSION: Fetal aortic time-domain measurement of volume flow is significantly increased in cases of fetal anemia due to red-cell alloimmunization. These findings can be used to improve the sensitivity, specificity and positive predictive value of the non invasive techniques used to predict fetal anemia, and may help in the selection of pregnancies that require cordocentesis and transfusion. PMID- 15133793 TI - Umbilical cord occlusion of the donor versus recipient fetus in twin-twin transfusion syndrome. AB - OBJECTIVES: To compare the technical characteristics and perinatal outcomes of selected twin-twin transfusion syndrome (TTTS) cases treated with primary umbilical cord occlusion (UCO) of the donor vs. recipient twin. METHODS: TTTS patients who underwent primary UCO via ultrasound endoscopic-guided umbilical cord ligation (UCL) or endoscopic umbilical cord photocoagulation (UCP) were eligible for the study. Primary UCO was offered only if a discordant lethal anomaly was present or by patient choice in TTTS Stages III/IV on the cord of the fetus with abnormal Doppler studies or hydrops. RESULTS: Primary UCO was performed in 25 TTTS cases between July 1997 and June 2002. UCO was performed in six (24%) donor twins and 19 (76%) recipient twins. UCL was performed in 23 cases and UCP in two cases. Overall, 22 (88%) patients had liveborn infants. There were no differences in cotwin survival (100% vs. 84.2%), median gestational age at delivery (34.8 vs. 33.8 weeks) and preterm premature rupture of membranes rate (16.6% vs. 15.7%) whether the donor or recipient twin, respectively, was the subject of UCO. However, two-trocar access (50% vs. 5.3%; P = 0.03) and amnioinfusion (83.3% vs. 0%; P < 0.001) were required more often in the UCO of the donor than of the recipient twin, respectively. Operating time (75 vs. 40 min) was significantly longer in UCO of the donor twin (P = 0.04). UCP was more likely to be used in occlusion of the donor's cord (33%) than of the recipient's cord (0%; P = 0.05). CONCLUSION: Primary UCO of the donor twin is technically more demanding than that of the recipient twin, but perinatal outcomes of the cotwins are comparable. PMID- 15133792 TI - Fetal middle cerebral artery peak systolic velocity in the investigation of non immune hydrops. AB - OBJECTIVE: In some cases of non-immune hydrops there is congenital or acquired fetal anemia. The aim of this study was to investigate the potential value of fetal middle cerebral artery peak systolic velocity (MCA-PSV) in the assessment and management of non-immune hydrops due to anemia. METHODS: Fetal MCA-PSV and fetal hemoglobin concentration, in blood obtained by cordocentesis, were measured in 16 singleton pregnancies referred to our unit for further investigations because of a diagnosis of non-immune hydrops fetalis. In all cases a detailed ultrasound examination demonstrated moderate or severe ascites, with or without skin edema, and pericardial or pleural effusions. Furthermore, there were no obvious malformations to account for the hydrops. In each fetus the measured MCA PSV and hemoglobin concentration were expressed as delta values (the difference in SD from the normal mean for gestation). Regression analysis was used to determine the significance of the association between delta MCA-PSV and delta fetal hemoglobin concentration. In addition, we searched our database to identify the sonographic features and hemoglobin concentration of fetuses with congenital infection. RESULTS: In the 16 cases of non-immune hydrops there were seven with parvovirus B19 infection, one each of alpha-thalassemia and primary cardiomyopathy and seven with no obvious explanation for the hydrops. There was a significant association between delta MCA-PSV and delta hemoglobin concentration (delta hemoglobin = (delta MCA-PSV + 0.1437)/-0.4154; R(2) = 0.7202; P < 0.0001). In 10 of the cases the fetal hemoglobin concentration was more than 4 SD below the normal mean for gestation and in all these cases the MCA-PSV was more than 2 SD above the normal mean for gestation. Our computer search identified an additional nine fetuses with parvovirus B19 infection and in all cases the predominant sonographic finding was ascites and the hemoglobin concentration was more than 4 SD below the normal mean. In contrast, only 3/14 fetuses with cytomegalovirus, toxoplasmosis, coxsackie B or Treponema infection had ascites and only 2/14 had a hemoglobin deficit of 4-6 SD. CONCLUSION: In the management of non-immune hydrops, measurement of fetal MCA-PSV can help identify the subgroup with fetal anemia. PMID- 15133794 TI - Tailored management of twin reversed arterial perfusion (TRAP) sequence. AB - OBJECTIVE: To describe our management of pregnancies complicated by twin reversed arterial perfusion (TRAP) sequence. METHODS: This was a retrospective study involving all cases of TRAP sequence referred to our fetal medicine unit in a 3 year period (2000-2002). Patients were routinely managed by repeat sonographic surveillance with sonographic anatomical evaluation and detailed echocardiography. Cases with signs of impending cardiac failure were treated by in-utero YAG-laser coagulation of the umbilical vessels of the acardiac twin. RESULTS: Six cases were studied. Three patients in whom there were no signs of deterioration in the status of the pump twin, and in whom the acardiac twin was smaller than the pump twin, were managed conservatively. However, one of these with monoamniotic twins ended in intrauterine fetal death of the pump twin. The other two cases presented with spontaneous cessation of blood flow in the umbilical artery of the acardiac twin. Both delivered at term normal neonates whose follow-up revealed no signs of neurological sequelae. One case of quadruplet pregnancy (with TRAP sequence and two dichorionic twins) was treated by selective termination of the monochorionic twins. Two cases with signs of impending cardiac failure were treated by in-utero YAG-laser occlusion of the vessels in the acardiac mass. Both interventions had a favorable outcome. CONCLUSIONS: Conservative treatment is suitable for milder cases of TRAP sequence in which the pump twin is the larger one. Cases in which the acardiac twin is larger have a poorer prognosis and should be treated by invasive intervention and cord occlusion. PMID- 15133795 TI - Incidence and characteristics of umbilical artery intermittent absent and/or reversed end-diastolic flow in complicated and uncomplicated monochorionic twin pregnancies. AB - OBJECTIVE: To evaluate the incidence and clinical relevance of intermittent absent and/or reversed diastolic flow on umbilical artery Doppler in different groups of monochorionic twin pregnancies. METHODS: This was a prospective study involving three groups of monochorionic pregnancies: Group 1: controls followed fortnightly from the first trimester (n = 80); Group 2: cases with selective intrauterine growth restriction (n = 40); and Group 3: cases with severe twin twin transfusion syndrome (n = 50). The presence and persistence over time of intermittent absent and/or reversed end-diastolic flow on umbilical artery Doppler was recorded. Placentas were examined and placental sharing and the presence of large arterioarterial anastomoses (AAA) was assessed. Perinatal outcome was recorded in all cases. RESULTS: Intermittent absent and/or reversed diastolic flow was present in 5% (4/80) of cases in Group 1, 45% (18/40) in Group 2 and 2% (1/50) in Group 3 (P < 0.0001, Group 2 vs. 1 and 3). Placental examination was performed in 76.4% (130/170) of cases and sharing was 58% for Group 1, 81% for Group 2 and 73% for Group 3 (P < 0.0001, Groups 2 and 3 vs. 1). Large AAA were identified in all examined cases with intermittent flow (18/18) and in 3.6% (4/112) of those without. The in-utero mortality rate was 0% in Group 1 and in Group 2 fetuses without intermittent flow. However, it was 19.4% in Group 2 cases with intermittent diastolic flow. CONCLUSIONS: Intermittent absent and/or reversed end-diastolic flow may be considered to be a characteristic sign of monochorionic pregnancy, and seems to result from the existence of large AAA. Its incidence is significantly increased in the context of selective intrauterine growth restriction, indicating a high risk for poor pregnancy outcome in these cases. PMID- 15133796 TI - Fetal heart rate and umbilical artery flow velocity variability in intrauterine growth restriction: a matched controlled study. AB - OBJECTIVES: To study heart rate and umbilical artery blood flow velocity variability in growth-restricted fetuses and investigate the influence of the autonomic nervous system on these parameters. METHODS: Doppler velocity waveforms were collected from long-lasting umbilical artery recordings in 15 fetuses with growth restriction and 15 normal age-matched controls at 23-35 weeks of gestation. Absolute heart rate and umbilical artery blood flow velocity as well as the coefficient of variation were determined. Using power spectral analysis the low- and high-frequency bands of heart rate variability and blood flow velocity variability were calculated. The low-to-high (LH) ratio of heart rate variability and blood flow velocity variability were examined as a measure of sympathovagal balance. RESULTS: In growth-restricted fetuses umbilical artery velocities were significantly reduced. Heart rate variability was significantly reduced in the presence of growth restriction, but no significant difference was demonstrated for blood flow velocity variability. The LH ratio for heart rate variability was significantly decreased in growth restriction, but no difference in LH ratio was demonstrated for blood flow velocity variability. CONCLUSION: Flow velocity variability in growth restriction seems not to be predominantly influenced by the autonomic nervous system, whereas the decreased heart rate variability seems to be influenced by altered sympathetic-parasympathetic balance. PMID- 15133797 TI - Does the availability of maternal HbA1c results improve the accuracy of sonographic diagnosis of macrosomia? AB - OBJECTIVE: To determine whether measuring maternal glycosylated hemoglobin (HbA1c) can improve the accuracy of sonographic estimation of fetal macrosomia. METHODS: Sonographic estimation of fetal weight (EFW) and maternal HbA1c were obtained in term, non-diabetic patients within 1 week before delivery. Neonatal birth weights were recorded at delivery and compared with both sonographic estimations and HbA1c. Macrosomia was defined as birth weight of >or=4000 g. The absolute error of the sonographic EFW was calculated. Receiver-operating characteristics (ROC) curve analysis was used to evaluate sonographic EFW and HbA1c as predictors of birth weight >or=4000 g. Variables were tested using regression analysis and student's t-test. RESULTS: One hundred and sixty two patients were evaluated between July and December 2002. Twenty-eight patients (17.3%) delivered macrosomic infants. Sonographic EFW >or=4000 g predicted macrosomia with sensitivity, specificity and positive and negative predictive values of 66.6%, 88.8%, 54.5% and 93.0%, respectively. Its overall accuracy was 85.5%. The area under the ROC curve of sonographic EFW in the prediction of macrosomia was 0.9 (P < 0.001). HbA1c levels in women delivering macrosomic and non-macrosomic neonates were 5.3 +/- 0.7% and 5.2 +/- 0.5%, respectively (P = 0.27). The area under the ROC curve of HbA1c in the prediction of macrosomia was 0.53 (P = 0.27). CONCLUSIONS: Maternal HbA1c is not a useful test in the prediction of birth weight. It therefore cannot be used to improve the accuracy of sonographic EFW. PMID- 15133798 TI - Ultrasound diagnosis of severe thrombotic placental damage in the second trimester: an observational study. AB - OBJECTIVES: To screen women with uteroplacental insufficiency between 18 and 26 weeks' gestation for sonographic evidence of destructive placental lesions, to observe the effect of low molecular-weight heparin (LMWH) in these cases, and to compare the outcome with similar but untreated controls. METHODS: We screened 180 women at high risk for placental damage using 16-week maternal serum screening (alpha-fetoprotein and human chorionic gonadotropin), placental shape and texture, and uterine artery Doppler waveforms at the 18-20-week level II examination. Serial gray-scale examinations of placental texture were performed at 22, 24 and 26 weeks. LMWH was offered to women with ultrasound evidence of destructive placental lesions in the absence of intrauterine growth restriction and/or pre-eclampsia. RESULTS: We prospectively identified six women (3.3%) with abnormal maternal serum screening and uterine artery Doppler in whom abnormal placental texture (echogenic cystic lesions) suggestive of destructive lesions in the placental parenchyma was found either at the 18-20-week ultrasound examination (n = 4), or by 26 weeks of gestation (n = 2). All six received LMWH and had live births (gestational age at delivery, 33-37 weeks; birth weight, 1000 3200 g). A further 14 women were referred with similar multiparameter evidence of placental damage at or after 26 weeks, outside the screening study. All had significant fetal growth restriction and were therefore not offered heparin. In 9/14 cases there was a perinatal death. Ischemic and/or thrombotic placental pathology was confirmed in each case, but no maternal thrombophilia disorders were identified in the 20 women. CONCLUSIONS: Integrated biochemical and ultrasound testing of placental function at 16-20 weeks of gestation, followed by serial placental gray-scale ultrasound, may be an effective method of identifying a subset of pregnancies at high risk of adverse pregnancy outcome due to destructive lesions in the placental parenchyma. This strategy of identifying thrombo-occlusive placental lesions before the development of pregnancy complications may prove useful in the design of trials to study the effectiveness of LMWH in the prevention of clinical complications resulting from thrombo occlusive placental disease. PMID- 15133799 TI - Comparison of pregnancy outcome of euploid fetuses with increased nuchal translucency (NT) expressed in NT MoM or delta-NT. AB - OBJECTIVE: To evaluate the outcome of euploid fetuses with increased nuchal translucency thickness (NT) expressed in multiples of the median (MoM) or delta NT. METHODS: Included in the study were euploid fetuses with increased NT >or= 95(th) centile, for which information about pregnancy outcome was available. The following parameters were defined as an adverse outcome: miscarriage, structural anomalies justifying termination of pregnancy, and structural anomalies, genetic syndromes and neurodevelopmental problems diagnosed postnatally. Fetal outcome according to NT MoM and delta-NT was calculated using different cut-off values. Calculations of the odds ratio for adverse outcome were performed using either NT MoM or delta-NT as a predictor in logistic regression models. RESULTS: The study comprised 168 euploid fetuses. Of these, 38 (23%) had an adverse outcome: 11 (6%) had miscarriages, 14 (8%) were terminated because of fetal abnormalities detected on the prenatal scan and 13 (7%) were found postnatally to have abnormalities. The incidence of cases exhibiting an adverse outcome was 5.3%, 19.2% and 58.5% for NT values of 1.6-1.9, 2.0-3.0 and >3.0 MoM, respectively (P < 0.0001, chi(2) test), and 3.9%, 16.7% and 62.8% for delta-NT values of 1.0-1.4, 1.5-2.5 and >2.5 mm, respectively (P < 0.0001, chi(2) test). Using cut-offs of 2.0 MoM and delta NT of 1.5 mm, the odds ratios for adverse outcome were 10.2 (95% CI, 3.4-30.4) and 15.4 (95% CI, 4.2-43.6), respectively. CONCLUSION: Both the NT MoM and delta NT approaches may be used to determine cases which require additional antenatal investigation as well as fetal karyotyping. For this purpose we suggest using a cut-off of either 2.0 MoM or a delta-NT of 1.5 mm. PMID- 15133800 TI - Early prenatal diagnosis by celocentesis. AB - BACKGROUND: Celocentesis is the ultrasound-guided aspiration of fluid from the extra-amniotic cavity at 7-8 weeks of gestation. This paper reports on the clinical application of celocentesis for early prenatal diagnosis. METHODS: Celocentesis was successfully performed in nine pregnancies and 1-2 mL of fluid were obtained after one needle insertion. The indications were prenatal diagnosis of beta-thalassemia or sickle cell disease (n = 6), Marfan syndrome (n = 1) and paternity testing (n = 2). Molecular biological techniques were used to analyze the celomic fluid and this was successfully carried out in all cases. RESULTS: In two cases pregnancy termination was performed at the request of the mother because in one case the fetus was found to have sickle cell anemia and in the second case paternity testing demonstrated that the father was not the woman's husband. In both cases the results were confirmed using the placental samples collected after pregnancy termination. In six of the seven pregnancies with desirable results, amniocentesis was performed at 16 weeks and the results were concordant with those obtained from celocentesis. All pregnancies were uneventful and resulted in the delivery of healthy and appropriately grown babies. CONCLUSION: Celocentesis may be a viable alternative to the currently used tests of chorionic villus sampling and amniocentesis. PMID- 15133801 TI - Lost intrauterine devices during pregnancy: maternal and fetal outcome after ultrasound-guided extraction. An analysis of 82 cases. AB - OBJECTIVES: It is generally agreed that intrauterine devices (IUDs) with visible strings in pregnancy should be removed because of the increased risk of miscarriage, septic complications and premature delivery. The precise management of pregnancies in association with so-called 'lost IUDs', and especially the technique of their removal, has remained controversial. We present our experience of the management of intrauterine pregnancies with a lost IUD. METHODS: Ultrasound-guided extraction of a lost IUD was performed in 82 intrauterine pregnancies. The subsequent outcome of the pregnancies is described. RESULTS: There were no intra- or post-procedure maternal complications. Although the miscarriage rate in the first 3 weeks after the procedure was higher than that in normal pregnancy, the complication rate approached that of normal pregnancy as the pregnancies progressed. The total miscarriage rate of 22% was comparable to that following extraction of IUDs with visible filaments. The rate of live births was 77.0%. Delivery before 37 weeks occurred in 13.5% of cases. CONCLUSIONS: Ultrasound-guided extraction is a minimally invasive and inexpensive procedure that is associated with few postoperative complications. It has a high success rate and is associated with a moderate miscarriage rate and no maternal complications. PMID- 15133802 TI - Sonographic findings in acute urinary retention secondary to retroverted gravid uterus: pathophysiology and preventive measures. AB - OBJECTIVES: To explore the pathophysiology of acute urinary retention in women with a retroverted gravid uterus and to suggest measures to prevent its recurrence. METHODS: In five women with a retroverted gravid uterus and acute urinary retention necessitating catheterization, the morphology of the genitourinary system was assessed by using transabdominal, transvaginal and introital sonography. RESULTS: In the supine resting position, the cervix was displaced superiorly and anteriorly by the impacted and retroverted uterus so that it compressed the lower bladder, leading to obstruction of the internal urethral orifice. The upper bladder extended superiorly and overlay the uterus. During straining, urethral motion was not limited and there was an average rotational angle of the bladder neck of 32 degrees, ranging from 21 degrees to 44 degrees. Increasing abdominal pressure further compressed the lower bladder. Measures suggested to the women for the prevention of urinary retention included limiting fluid intake before sleep, changing from the supine to the prone position before getting up and avoiding a Valsalva maneuver but performing a Crede maneuver during voiding. In all except one case these measures successfully prevented recurrence. CONCLUSIONS: Acute urinary retention secondary to a retroverted gravid uterus is caused by a displaced cervix compressing the lower bladder and interfering with drainage to the urethra. The urethra itself is not compressed or distorted. Understanding the pathophysiology of the lower urinary tract may allow maneuvers which prevent acute urinary retention. PMID- 15133803 TI - Assessment of changes in utero-ovarian arterial impedance during the peri implantation period by Doppler sonography in women undergoing assisted reproduction. AB - OBJECTIVE: To investigate changes in utero-ovarian blood flow during the peri implantation period and their significance in successful embryo implantation. METHODS: A prospective longitudinal study was conducted in 317 women undergoing in-vitro fertilization-embryo transfer (IVF-ET) treatment. All of them had at least one good-quality embryo for transfer on the second or third day after oocyte retrieval. Measurement of endometrial thickness and color flow imaging with pulsed waveform analysis of uterine and ovarian arteries were performed before ET and 5-6 days after ET. RESULTS: There were no significant differences in the age of patients, duration of infertility or number of embryos transferred between women who became pregnant (n = 91) and those who did not (n = 226). There was no difference in mean endometrial thickness between the two groups before ET, while a thicker endometrium was found in women who had conceived compared with those who had not 5-6 days after ET (P = 0.02). Mean uterine arterial resistance index (RI) and pulsatility index (PI) values were significantly lower in the pregnant than in the non-pregnant group before ET (P = 0.04 and P = 0.003, respectively), but no significant differences were found between the two groups 5 6 days after ET. In contrast, the mean ovarian arterial RI and PI values were similar between the two groups before ET, yet the pregnant group showed significantly lower RI and PI values compared with the non-pregnant group 5-6 days after ET (P = 0.002 and P = 0.01, respectively). A significantly higher peak systolic velocity (PSV) of intraovarian vessels was also noted in the pregnant group 5-6 days after ET. CONCLUSION: Different utero-ovarian blood flow changes during the peri-implantation period occur in conception and non-conception cycles in women following IVF. Doppler assessment of uterine arterial resistance can help to determine a time interval within the menstrual cycle that is of optimal endometrial status for embryo implantation in assisted conception programs. Delay in achieving adequate uterine perfusion during the temporal window of embryo implantation may have an impact on endometrial receptivity. PMID- 15133804 TI - The interobserver reliability of three-dimensional power Doppler data acquisition within the female pelvis. AB - OBJECTIVES: To examine the interobserver reliability of three-dimensional (3D) power Doppler data acquisition from the uterus and ovary. METHODS: 3D power Doppler angiography was used to acquire endometrial data from 20 patients and ovarian data from a further 20 different patients at various stages of in vitro fertilization. Two different observers each acquired two datasets from all 40 patients resulting in 80 endometrial and 80 ovarian datasets in total. Virtual Organ Computer-aided AnaLysis was used to define the object of interest and semiquantify the power Doppler signal within it. The reliability of measurements of volume and vascularity was assessed by calculating interclass correlation coefficients (ICCs) and limits of agreement between the two observers. RESULTS: Whilst volumetric data proved more reliably acquirable than power Doppler data, the interobserver reliability of data acquisition was high overall for both the endometrium and ovary with all measurements obtaining a lower ICC of above 0.9. Limits of agreement revealed minimal disagreement between the two observers for measurements of volume and vascularity within both the ovary and endometrium. The 'indices of vascularity' within the endometrium were less than those derived from the ovary and subendometrium, which were comparable. CONCLUSIONS: 3D ultrasound can be reliably used between observers to acquire power Doppler information from the ovary and endometrium thus supporting the current use and further development of this technique in clinical practice. PMID- 15133805 TI - Successful intrauterine treatment with radiofrequency ablation in a case of acardiac twin pregnancy complicated with a hydropic pump twin. AB - Twin reversed arterial perfusion sequence is a serious complication of monochorionic twin pregnancy, as the pump twin that perfuses blood to the acardiac twin may experience heart failure and fetal hydrops resulting in a poor perinatal outcome. A woman with an acardiac twin pregnancy complicated by a hydropic pump twin underwent intrauterine treatment with radiofrequency ablation (RFA) at 27 weeks of gestation. Obliteration of blood flow to the acardiac twin from the pump twin was successful. Fetal hydrops resolved by the time of delivery at 32 weeks of gestation, in spite of transient deterioration, and a good postnatal outcome was achieved for the pump twin. We found that RFA was an effective intrauterine treatment for acardiac twin pregnancy and suggest that it could be introduced in cases complicated by a hydropic pump twin. PMID- 15133806 TI - Prenatal diagnosis of intrauterine premature closure of the ductus arteriosus following maternal diclofenac application. AB - We report a case of prenatal diagnosis of premature closure of the ductus arteriosus following maternal diclofenac therapy at 35 weeks of gestation. Fetal echocardiography at 37 weeks of gestation revealed a dilated right ventricle with moderate tricuspid regurgitation, pulmonary insufficiency and complete closure of the ductus arteriosus. Immediate Cesarean section resulted in an excellent neonatal outcome. Whereas the effect of indomethacin on prenatal ductal constriction is well known, widely prescribed non-steroidal anti-inflammatory drugs such as diclofenac can have an equally deleterious effect and are best avoided in the third trimester of pregnancy. PMID- 15133807 TI - Idiopathic constriction of the fetal ductus arteriosus. AB - Constriction of the ductus arteriosus in the fetus is well studied, but mostly secondary to maternal exposure to indomethacin or other non-steroidal anti inflammatory medications, or structural cardiac lesions. We present a case of a fetus presenting with right ventricular hypertrophy secondary to an S-shaped ductus arteriosus with distal constriction diagnosed by pulsed Doppler imaging. A review of the recent English literature of similar cases is presented. PMID- 15133808 TI - Congenital constriction band of the upper arm: the role of three-dimensional ultrasound in diagnosis, counseling and multidisciplinary consultation. AB - Constriction band syndrome represents a sporadic condition that may result in amputations, constrictions and other deformities of the fetal limbs and body. Prenatal diagnosis by two-dimensional ultrasound has been reported. We present a case of constriction band involving the upper arm in which the assessment by three-dimensional ultrasound significantly contributed to the diagnosis and the multidisciplinary counseling. In fact, multiplanar imaging and surface rendering allowed a clear depiction of the extent of the constriction, the club-hand deformity and the relationship between the amniotic band, the cord and the fetal limb. This case represents a unique and effective application of three dimensional ultrasound in prenatal diagnosis. PMID- 15133809 TI - Pouch sign in prenatal diagnosis of esophageal atresia. PMID- 15133810 TI - Detection of increased middle cerebral artery peak systolic velocity in fetuses affected by hemoglobin H Quong Sze disease. PMID- 15133811 TI - A "cure" for Parkinson's disease: can neuroprotection be proven with current trial designs? AB - Current medical and surgical therapies for Parkinson's disease provide symptomatic control of motor impairments rather than slowing or halting the progression of the disease. Previous clinical trials examining drugs such as dopamine agonists and selegiline for neuroprotective effects used "surrogate" outcomes, including clinical measures (rating scales, time to require levodopa), neuroimaging techniques (beta-CIT single photon emission computed tomography; fluorodopa positron emission tomography), and mortality tracking. These studies failed to provide conclusive results because of design faults such as failing to control for symptomatic effects, small sample size, and not accounting for the possible effects of drugs on radionuclide tracer handling. Lessons must be learned from these failed neuroprotection trials. This review summarises the problems with previous neuroprotection studies and makes recommendations for future trial design. It is concluded that the primary outcome of explanatory trials should continue to be clinical measures such as the Unified Parkinson's Disease Rating Scale (UPDRS). It should be assumed that all agents have a symptomatic effect, which necessitates evaluation after a prolonged drug washout period. To achieve the evaluation after a prolonged drug washout period more effectively, trials must be performed in early disease and over a short period (6 12 months) so that symptomatic therapy is not required. To achieve adequate statistical power, these trials will need to include thousands of patients. Radionuclide imaging can only be used in such trials after considerable methodological work has been performed to establish its validity and reliability. To be affordable, such large explanatory trials need more streamlined designs with fewer hospital visits, fewer outcome measures, and rationalised safety monitoring. The clinical effectiveness of promising compounds from explanatory trials will need to be established in large long-term pragmatic trials using outcome measures such as quality of life, cost-effectiveness, and mortality. Such pragmatic trials could be continuations of the explanatory trials: after the primary outcome of the explanatory study (e.g., UPDRS) has been reported in an interim analysis, the trial could be continued for a further 5 to 10 years to report on quality of life and health economics outcomes. PMID- 15133812 TI - Semiquantitative study of current coffee, caffeine, and ethanol intake in essential tremor cases and controls. AB - There are several reasons to study caffeine, coffee, and ethanol intake in essential tremor (ET) patients. ET patients also might modify their use of these beverages because of their effects on tremor. Intake of caffeine, coffee, and ethanol has not been quantified in a group of ET patients. Our objective is to use a semiquantitative food frequency questionnaire to compare current daily intake of coffee, caffeine, and ethanol in ET patients and controls. A total of 130 ET cases were patients at the Neurological Institute of New York, and 175 controls were ascertained by random digit dialing. Caffeine (in milligrams) and ethanol (in grams) intake were calculated from a semiquantitative food-frequency questionnaire. Mean daily caffeine intake in patients was 138.4 versus 246.6 mg in controls; medians were 101.1 versus 175.5 mg (P < 0.001). Mean daily ethanol intake in patients was 8.2 versus 6.2 gm in controls; medians were 2.4 versus 1.9 gm (P = 0.89). Cases drank less coffee than controls, but drank similar amounts of tea, soft drinks, fruit juices, and milk. Daily caffeine intake was not correlated with tremor severity or duration. ET patients consumed less caffeine than did controls, which is likely to be a dietary modification in response to tremor. The observation that caffeine consumption was not correlated with tremor severity raises the additional possibility that lower caffeine consumption in ET patients may not exclusively be a response to tremor. A prospective study is needed to explore whether decreased caffeine consumption is a risk factor for ET. PMID- 15133813 TI - Enhancing recognition of early Parkinsonism in the community. AB - Because Parkinsonism is underdiagnosed in the community, we have validated screening modalities in the field setting and developed a screening procedure to enhance recognition of undiagnosed patients. In a first survey, we identified suspect cases among patients consulting 9 general practitioners (GPs) over a 3 week period using in parallel: (1) a published questionnaire; (2) a standardized examination by the GPs; (3) clinical impression of the GPs; or (4) pre established diagnoses. Parkinsonism was ascertained by two neurologists with a 1 year follow-up and FP-CIT-SPECT. In total, 1,411 patients consulted the GPs, 1,030 participated in the study, 87 possible cases were identified by at least one of four screening modalities, 12 suffered from Parkinsonism, and 4 of these 12 were de novo cases. Statistical analysis demonstrated that with appropriate evaluation, the questionnaire is highly sensitive and excludes most nonaffected persons, and that the GPs' clinical impression is more specific. We therefore tested in a second survey the efficacy of a serial screening, starting with the questionnaire, followed by a standardized GP evaluation, and then by neurological examination. Of 1,353 participants seen by 9 GPs during a 3-week period, 5 de novo cases were identified. This simple screening protocol significantly enhances recognition of incipient Parkinsonism. PMID- 15133814 TI - Drug adherence in Parkinson's disease. AB - Physicians modify drug schedules in response to their patients' clinical responses. Failure to relieve patients' symptoms or the emergence of drug-related side effects may reflect nonadherence to a prescribed drug schedule rather than incorrect therapeutic physician decisions. Using a medication questionnaire and a computerized medication event monitoring system (MEMS) to monitor medication use, nonadherence of drug use was examined in subjects with Parkinson's disease (PD). We report that prescription nonadherence in PD subjects was common and approximated that reported in other chronic diseases. During a 28-day observation period, only 4 of 39 subjects had complete schedule adherence, i.e., no missed, extra, or mistimed doses. Using a questionnaire, 24.3% of subjects acknowledged missing any doses but the computerized MEMS recorded that 51.3% of subjects missed at least one dose per week and 20.5% of subjects missed three or more doses per week. Mistiming of doses was admitted by 73% of subjects but 82.1% had recorded mistimed doses. Of multiple sociodemographic and disease-related items examined, only gender and level of education were statistically related to nonadherence. PMID- 15133815 TI - Measurements of transcallosally mediated cortical inhibition for differentiating parkinsonian syndromes. AB - Clinicopathologic evidence suggests differential involvement of cortex and corpus callosum (CC) in various disorders presenting with a parkinsonian syndrome. We tested the hypothesis of whether neurophysiologic and morphometric assessments of CC as surrogate parameters of cortical involvement could be helpful in differential diagnosis of parkinsonian disorders. The integrity of CC was assessed neurophysiologically by measuring the ipsilateral silent period (iSP) evoked by transcranial magnetic stimulation (TMS) in a total of 25 patients with idiopathic parkinsonian syndromes (IPS), corticobasal ganglionic degeneration (CBD), progressive supranuclear palsy (PSP), or multiple system atrophy (MSA). Additionally, morphometric analyses of magnetic resonance imaging (MRI) measurements of CC was carried out in all patients. iSP was abnormal in all 5 CBD and all 5 PSP patients, whereas it was intact in all 10 IPS patients and all 5 MSA patients. Among various MRI parameters of CC, testing between different groups revealed a significant difference only for measurements of the middle part of the truncus. CBD and PSP patients exhibited a significant atrophy as compared with control subjects. These data suggest impairment of callosal integrity in patients with CBD and PSP. iSP measurements may be a useful clinical neurophysiologic test in differential diagnosis of patients with parkinsonian syndromes. PMID- 15133816 TI - Increased risk of head tremor in women with essential tremor: longitudinal data from the Rochester Epidemiology Project. AB - In one cross-sectional study of a community in northern Manhattan, women with essential tremor (ET) were more likely to have head tremor than were men. In that study, patients were seen at one point in time, rather than followed longitudinally. Head tremor often develops after arm tremor, and its appearance in patients with ET may therefore be a function of duration of follow-up. In a second epidemiological study utilizing the Rochester Epidemiology Project, in which ET subjects were followed from disease diagnosis to death, we determined whether there was an association between female gender and head tremor. We utilized the records-linkage system of the Rochester Epidemiology Project to identify ET cases. Records were reviewed and clinical data abstracted by a neurologist specializing in movement disorders. A second neurologist reviewed a subsample of records. There were 107 ET cases (69 women, 38 men) followed for 10.1 +/- 9.1 years from ET diagnosis to death. Head tremor was present in 37 (53.6%) women and 5 (13.2%) men (odds ratio [OR] = 7.6, 95% confidence interval [CI] = 2.7-21.9, P < 0.001). In a multivariate linear regression analysis, women remained at high risk for head tremor (OR = 6.5, 95% CI = 2.2-19.0, P = 0.001) independent of disease duration. We found in this longitudinal epidemiological study that women with ET were six times more likely to develop head tremor over the course of their illness than were men. The reason for the association between gender and head tremor, which has now been demonstrated in several studies, is not known, but it could reflect gender differences in the distribution of disease pathology within the brain. PMID- 15133817 TI - Rapidly progressive behavioral changes and parkinsonism in a 68-year-old man. PMID- 15133818 TI - Mitochondrial complex I and IV activities in leukocytes from patients with parkin mutations. AB - The parkin protein functions as a RING-type ubiquitin protein ligase. Considering the possibility that impaired ubiquitin-proteosomal system activity may impair antioxidant defenses and enhance oxidative stress, we have investigated the activity of mitochondrial respiratory enzymes in patients with parkin gene mutations. A significant decrease in the leukocyte complex I activity was found both in patients with parkin mutations (62.5%) and idiopathic PD (64.5%) compared with age-matched controls (P < 0.001). Complex IV activity was also decreased significantly in idiopathic PD patients (60%), but no difference was detected between controls and patients with parkin mutations. PMID- 15133819 TI - Neuronal globus pallidus activity in patients with generalised dystonia. AB - We studied 516 globus pallidus neurons in dystonic patients. The firing rate was analysed. We classified the burst activity into tonic, burst, and pause patterns. Mean +/- SD firing rates and tonicity score for internal globus pallidus (GPi) and external globus pallidus (GPe) were 54.6 +/- 28.6; 58.01 +/- 39.1 and 1.18 +/ 0.55; 0.95 +/- 0.43, respectively. Differences in percentage appearance of tonic, burst, or paused neurons were not statistically significant for GPi versus GPe. GPi firing features in dystonic patients were closely similar to those of GPe. This could suggest that the abnormally patterned output from GPi would not result from increased differential inhibitory/excitatory input arising from the direct/indirect pathway but rather be transmitted from GPe, striatum, or either centromedian nucleus. PMID- 15133820 TI - Non-subtype-selective opioid receptor antagonism in treatment of levodopa-induced motor complications in Parkinson's disease. AB - Opioid peptide transmission is enhanced in the striatum of animal models and Parkinson's disease (PD) patients with levodopa-induced motor complications. Opioid receptor antagonists reduce levodopa-induced dyskinesia in primate models of PD; however, clinical trials to date have been inconclusive. A double-blind, placebo controlled, crossover design study in 14 patients with PD experiencing motor fluctuations was carried out, using the non-subtype-selective opioid receptor antagonist naloxone. Naloxone did not reduce levodopa-induced dyskinesia. The duration of action of levodopa was increased significantly by 17.5%. Non-subtype-selective opioid receptor antagonism may prove useful in the treatment of levodopa-related wearing-off in PD but not in dyskinesia. PMID- 15133821 TI - Grip force abnormalities in de novo Parkinson's disease. AB - In recent years it has been shown that a variety of movement disorders are associated with abnormalities of the fine motor control of the hand. In Parkinson's disease (PD), these changes consist of a slowing of the rate of grip force development and the use of abnormally large grip forces both during lifting and static holding of an object. It has been suggested, however, that these changes are a direct effect of the patient's levodopa medication or associated with levodopa induced dyskinesias. Accordingly, we examined the performance of de novo Parkinson patients in a precision lifting task. All patients (n = 6) were newly diagnosed and showed rigidity, bradykinesia, or both, but were unaffected by tremor or dyskinesia. None of the patients had received antiparkinson medication. Grip force was abnormally high in both the lifting and hold phases. This exaggeration was equal in magnitude to that observed previously in medicated patients. Thus we conclude that the abnormalities in grip force observed here are intrinsic features of PD and not the result of dopamine medication or its side effects. PMID- 15133822 TI - Verb and noun generation tasks in Huntington's disease. AB - We compared noun- and verb-generation tasks in a demented group (n = 9, Dementia Rating Scale < or = 129) and in a non-demented group (n = 17, Dementia Rating Scale > 129) of Huntington's disease (HD) patients compared to 26 matched normal subjects. We did not find a specific deficit for verb production in non-demented patients who had a performance similar to but weaker than that of the controls across the four tasks. The profile of results was different in the demented group because, apart from a global deficit whatever the task in comparison with both non-demented and control groups, the demented patients exhibited increased difficulties in the two tasks implying verb production. The deficit of verb production observed in demented HD patients is discussed in relation to the damage to the motor loop in HD patients at later stages of disease. PMID- 15133824 TI - Sporadic case of dentatorubral pallidoluysian atrophy with no CAG repeat expansion and no intranuclear inclusions. AB - We present a patient with a 14-year course beginning at the age of 44 years with hemidystonia followed by generalized choreoathetosis, behavioral, and oculomotor disturbances. Family history and genetic testing were unrevealing. Neuropathological findings were identical to genetic dentatorubral pallidoluysian (DRPLA) except for the lack of intranuclear inclusions. PMID- 15133823 TI - Cerebrospinal fluid analysis differentiates multiple system atrophy from Parkinson's disease. AB - We investigated whether cerebrospinal fluid (CSF) analysis discriminates between idiopathic Parkinson's disease (PD; n = 35) and multiple system atrophy (MSA; n = 30). The median CSF concentration of the neurotransmitter metabolites 5 hydroxyindolacetic acid (5-HIAA) and 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) was reduced significantly (49-70%) in MSA compared to PD. In contrast, several brain-specific proteins (tau, neuron-specific enolase, myelin basic protein) were elevated (130-230%) in MSA compared with those in PD. A combination of CSF tau and MHPG discriminated PD from MSA (adjusted odds ratios: tau, 27.2; MHPG, 0.14). Our data suggest that the more progressive and widespread neurodegenerative nature of MSA, as compared with PD, is reflected in the composition of CSF. We propose that CSF analysis may become part of the diagnostic work-up of patients with parkinsonian syndromes. PMID- 15133825 TI - Unilateral deep brain stimulation of the internal globus pallidus alleviates tardive dyskinesia. AB - We describe a patient with fluspirilene-induced tardive dyskinesia of the choreiform oro-facial-laryngeal type resistant to various conservative approaches for 7 years who underwent deep brain stimulation of the internal pallidal globe. We found immediate and marked suppression of her perioral involuntary movements with unilateral stimulation at 60 Hz. PMID- 15133826 TI - Dystonia as a presenting sign of spinocerebellar ataxia type 1. AB - We report on a 39-year-old man who presented initially with marked blepharospasm, oromandibular dystonia and retrocollis and one year later developed mild ataxia. Our findings suggest that dystonia can be a disabling presenting sign of SCA1 and support the clinical heterogeneity of SCA1, highlighting the importance of considering this entity in patients combining dystonia and cerebellar ataxia. PMID- 15133827 TI - Computed tomographically-controlled injection of botulinum toxin into the longus colli muscle in severe anterocollis. AB - We report on a 44-year-old man who suffered from severe anterocollis. Repeated computed tomographically controlled injections of botulinum toxin into the right longus colli muscle allowed a precise location of the needle and injection of the toxin, leading to clear improvement of symptoms. PMID- 15133828 TI - Arg(184)His mutant GTP cyclohydrolase I, causing recessive hyperphenylalaninemia, is responsible for dopa-responsive dystonia with parkinsonism: a case report. AB - We describe a 54-year-old man with dominant adult-onset dopa-responsive dystonia (DRD) with parkinsonism caused by an Arg184His mutation in guanosine 5' triphosphate cyclohydrolase I (GCH-I). This is the first mutation in the GCH-I gene that has been proven to be responsible for both recessive and dominant phenotypes. PMID- 15133829 TI - SCA2 presenting as levodopa-responsive parkinsonism in a young patient from the United Kingdom: a case report. AB - We report on a young woman from the United Kingdom with L-dopa-responsive parkinsonism with a trinucleotide repeat expansion in her spinocerebellar ataxia 2 (SCA2) gene. The case further extends the phenotype of SCA2 and emphasizes the importance of SCA screening in young-onset parkinsonism, irrespective of ethnic origin. PMID- 15133830 TI - Propriospinal myoclonus after treatment with ciprofloxacin. AB - The clinical and electrophysiological features of a truncal myoclonus in a 55 year-old man are described. The electromyographic characteristics point toward propriospinal myoclonus. It is suggested that a myoclonic generator was released after use of ciprofloxacin, by antagonising the gamma-aminobutyric acid metabolism. PMID- 15133831 TI - Senile chorea treated by deep brain stimulation: a clinical, neurophysiological and functional imaging study. AB - We report on a patient with senile chorea, treated with deep brain stimulation of the left globus pallidus internus and subsequently the left ventralis oralis posterior nucleus of the thalamus. Deep brain field potential recordings and functional imaging using single photon emission tomography enabled us to suggest pathophysiological mechanisms for the symptoms. PMID- 15133832 TI - Atypical parkinsonism and Annonaceae consumption in New Caledonia. PMID- 15133833 TI - Atypical parkinsonism in New Caledonia: comparison with Guadeloupe and association with Annonaceae consumption. PMID- 15133834 TI - Re: UPDRS: Status and recommendations. PMID- 15133835 TI - Do on-off variations cause discrepancies in the historical items of the UPDRS? PMID- 15133836 TI - Application of proteomics technologies in the investigation of the brain. AB - Approximately 30-50% of the genes in mammals are expressed in the nervous system. A differential expression of genes in distinct patterns is necessary for the generation of the large variety of neuronal phenotypes. Proteomic analysis of brain compartments may be useful to understand the complexity, to investigate disorders of the central nervous system, and to search for corresponding early markers. Up to now, proteomics has mainly studied the identity and levels of the abundant human, rat, and mouse brain proteins as well as changes of their levels and the modifications that result from various neurological disorders, like Alzheimer's disease and Down's syndrome in humans and in animal models of those diseases. The proteins, for which altered levels in these disorders have been observed, exert mainly neurotransmission, guidance, and signal-transduction functions, or are involved in detoxification, metabolism, and conformational changes. Some of those proteins may be potential drug targets. Further improvement of proteomics technologies to increase sensitivity and efficiency of detection of certain protein classes is necessary for a more detailed analysis of the brain proteome. In this review, a description of the proteomics technologies applied in the investigation of the brain, the major findings that resulted from their application, and the potential and limitations of the current technologies are discussed. PMID- 15133837 TI - Identification of protein associations in organelles, using mass spectrometry based proteomics. AB - Recent literature that highlights the power of using mass spectrometry (MS) for protein identification from preparations of highly purified organelles and other large subcellular structures is covered in this review with an emphasis on techniques that preserve the integrity of the functional protein complexes. Recent advances in distinguishing contaminant proteins from "bonafide" organelle localized proteins and the affinity capture of protein complexes are reviewed, as well as bioinformatic strategies to predict protein organellar localization and to integrate protein-protein interaction maps obtained from MS-affinity capture methods with data obtained from other techniques. Those developments demonstrate that a revolution in cellular biology, fueled by technical advances in MS-based proteomic techniques, is well underway. PMID- 15133838 TI - Mass spectrometry for detection of 4-hydroxy-trans-2-nonenal (HNE) adducts with peptides and proteins. AB - Despite the great technical advancement of mass spectrometry, this technique has contributed in a limited way to the discovery and quantitation of specific/precocious markers linked to free radical-mediated diseases. Unsaturated aldehydes generated by free radical-induced lipid peroxidation of polyunsaturated fatty acids, and in particular 4-hydroxy-trans-2 nonenal (HNE), are involved in the onset and progression of many pathologies such as cardiovascular (atherosclerosis, long-term complications of diabetes) and neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, and cerebral ischemia). Most of the biological effects of HNE are attributed to the capacity of HNE to react with the nucleophilic sites of proteins and peptides (other than nucleic acids), to form covalently modified biomolecules that can disrupt important cellular functions and induce mutations. By considering the emerging role of HNE in several human diseases, an unequivocal analytical approach as mass spectrometry to detect/elucidate the structure of protein-HNE adducts in biological matrices is strictly needed not only to understand the reaction mechanism of HNE, but also to gain a deeper insight into the pathological role of HNE. This with the aim to provide intermediate diagnostic biomarkers for human diseases. This review sheds focus on the "state-of-the-art" of mass spectrometric applications in the field of HNE-protein adducts characterization, starting from the fundamental early studies and discussing the different MS-based approaches that can provide detailed information on the mechanistic aspects of HNE-protein interaction. In the last decade, the increases in the accessible mass ranges of modern instruments and advances in ionization methods have made possible a fundamental improvement in the analysis of protein-HNE adducts by mass spectrometry, and in particular by matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) tandem mass spectrometry. The recent developments and uses of combined analytical approaches to detect and characterize the type/site of interaction have been highlighted, and several other aspects, including sample preparation methodologies, structure elucidation, and data analysis have also been considered. PMID- 15133840 TI - Alterations in expression, proteolysis and intracellular localizations of clusterin in esophageal squamous cell carcinoma. AB - AIM: To investigate biogenesis and intracellular localizations of clusterin to elucidate the potential molecular mechanisms implicated in tumorigenesis of esophageal mucosa. METHODS: Semi-quantitative RT-PCR for multi-region alteration analysis, Western blot for different transcriptional forms and immunohistochemical staining for intracellular localizations of clusterin were carried out in both tissues and cell lines of ESCC. RESULTS: The N-terminal deletions of the clusterin gene and the appearance of a 50-53 ku nuclear clusterin, an uncleaved, nonglycosylated, and disulfide-linked isoform, were the major alterations in cancer cells of esophagus. Naturally the 40 ku clusterin was located in the connective tissue of the lamina propria of epithelial mucosa and right under the basal membrane of epithelia, but it was disappeared in stromal mucosa of esophagus and the pre-matured clusterin was found positive in cancerous epithelia. CONCLUSION: The N-terminal deletion of clusterin may be essential for its alterations of biogenesis in ESCC. PMID- 15133841 TI - Effects of c9,t11-conjugated linoleic acid on adhesion of human gastric carcinoma cell line SGC-7901. AB - AIM: To investigate the effect of c9,t11-conjugated linoleic acid (c9,t11-CLA) on the adhesion of human gastric carcinoma cell line (SGC-7901). METHODS: SGC-7901 cells were at first treated with different concentrations (25, 50, 100, 200 micromol/L) of c9,t11-CLA and 1 mL/L ethanol (as a negative control) for 24 h. Using adhesion assay and Western blot, we investigated the ability of SGC-7901 cells to adhere to intracellular matrix and examined the expression of E-cadherin (ECD), alpha-catenin, intercellular adhesion molecule 1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) in these cells. RESULTS: The attachment rate to laminin of SGC-7901 cells treated with different concentrations of c9,t11-CLA (0, 25, 50, 100, and 200 micromol/L) was 100.0+/-3.3, 95.7+/-4.0, 89.2+/-4.6, 87.9+/ 6.1, and 65.9+/-5.8, respectively. The attachment rate to fibronectin was 100.0+/ 4.7, 96.8+/-3.8, 94.5+/-4.1, 76.5+/-4.3, and 61.8+/-4.8, respectively. The attachment rate to Matrigel was 99.9+/-6.6, 91.4+/-6.8, 85.5+/-7.4, 79.3+/-5.6, and 69.6+/-5.1, respectively. Besides, c9,t11-CLA could increase the level of ECD and alpha-catenin, and decrease the level of ICAM-1 and VCAM-1 in SGC-7901 cells. CONCLUSION: c9,t11-CLA can reduce the adhesion of human gastric carcinoma cells to laminin, fibronectin and Matrigel. c9,t11-CLA can increase the level of ECD and alpha-catenin, and decrease the level of ICAM-1 and VCAM-1 in human gastric carcinoma cells. PMID- 15133842 TI - Diagnosis and surgical treatments of hepatocellular carcinoma with tumor thrombosis in bile duct: experience of 34 patients. AB - AIM: Hepatocellular carcinoma (HCC) with bile duct tumor thrombosis (BDT) is a rare event. The prognosis of this type of patients is very dismal. The aim of this study was to share the experience in the diagnosis and treatment of HCC with BDT, to further improve the prognosis of these patients. METHODS: Thirty-four patients of HCC with BDT received surgical treatment in authors' institute from July 1987 to January 2003 were reviewed retrospectively. The experience in the diagnosis and treatment, and the outcome of this type of HCC patients were summarized. RESULTS: Thirty of the 34 patients (88.2%) were positive for alpha fetoprotein (AFP) (>20 microg/L), and 12 patients (35.3%) were found having obstructive jaundice before operation, 18 cases were suspected of "obstruction of bile duct" preoperatively. The primary tumors were frequently located at the left medial (13 cases) or right anterior lobe (14 cases). Thirty-one patients received liver resections and removal of BDT, while the other 3 patients received removal of BDT combined with hepatic artery ligation and cannulation (HAL+HAI), or only removal of BDT because their liver function reservation and general condition could not tolerate the primary tumor resection. The 1-year survival rate was 71.4%(20/28). The longest disease-free survival was over 15 years. The intrahepatic tumor recurrence within 1 year after operation was found in 14 patients (14/28, 50.0%). CONCLUSION: Surgical removal of primary tumors and BDT is safe and beneficial to the HCC patients with BDT. Early detection, diagnosis, and surgical treatment are the key points to prolong the survival time of patients. PMID- 15133843 TI - Construction of human liver cancer vascular endothelium cDNA expression library and screening of the endothelium-associated antigen genes. AB - AIM: To gain tumor endothelium associated antigen genes from human liver cancer vascular endothelial cells (HLCVECs) cDNA expression library, so as to find some new possible targets for the diagnosis and therapy of liver tumor. METHODS: HLCVECs were isolated and purified from a fresh hepatocellular carcinoma tissue sample, and were cultured and proliferated in vitro. A cDNA expression library was constructed with the mRNA extracted from HLCVECs. Anti-sera were prepared from immunized BALB/c mice through subcutaneous injection with high dose of fixed HLCVECs, and were then tested for their specificity against HLCVECs and angiogenic effects in vitro, such as inhibiting proliferation and inducing apoptosis of tumor endothelial cells, using immunocytochemistry, immunofluorescence, cell cycle analysis and MTT assays, etc. The identified xenogeneic sera from immunized mice were employed to screen the library of HLCVECs by modified serological analyses of recombinant cDNA expression libraries (SEREX). The positive clones were sequenced and analyzed by bio-informatics. RESULTS: The primary cDNA library consisted of 2 x 10(6) recombinants. Thirty-six positive clones were obtained from 6 x 10(5) independent clones by immunoscreening. Bio-informatics analysis of cDNA sequences indicated that 36 positive clones represented 18 different genes. Among them, 3 were new genes previously unreported, 2 of which were hypothetical genes. The other 15 were already known ones. Series analysis of gene expression (SAGE) database showed that ERP70, GRP58, GAPDH, SSB, S100A6, BMP-6, DVS27, HSP70 and NAC alpha in these genes were associated with endothelium and angiogenesis, but their effects on HLCVECs were still unclear. GAPDH, S100A6, BMP-6 and hsp70 were identified by SEREX in other tumor cDNA expression libraries. CONCLUSION: By screening of HLCVECs cDNA expression library using sera from immunized mice with HLCVECs, the functional genes associated with tumor endothelium or angiogenesis were identified. The modified SEREX, xenogeneic functional serum screening, was demonstrated to be effective for isolation and identification of antigen genes of tumor endothelium, and also for other tumor cell antigen genes. These antigen genes obtained in this study could be a valuable resource for basic and clinical studies of tumor angiogenesis, thus facilitating the development of anti- angiogenesis targeting therapy of tumors. PMID- 15133844 TI - Effects of endostatin-vascular endothelial growth inhibitor chimeric recombinant adenoviruses on antiangiogenesis. AB - AIM: To investigate the inhibitory effects of endostatin-vascular endothelial growth inhibitor (VEGI151) recombinant adenoviruses on neovascularization. METHODS: We used recombinant adenoviruses to treat human vascular endothelial cell line ECV304, human hepatocellular carcinoma cell line HepG2, and murine fibroblast cell line L929, in order to study the chimeric gene expression in these cell lines. Chick choriallantic membrane (CAM) model, rabbit inflammatory corneal neovascularization (CNV) model, and liver cancer-bearing nude mice model were employed to investigate the negative biological effect of fusion molecules on neovascularization in vivo. RESULTS: Western blot showed that the molecular weight of fusion protein was about 41 kD after infection of ECV304, HepG2 and L929 cells with supernatant of AdhENDO-VEGI151. The fusion protein showed a specific inhibitory effect on the proliferation of ECV304 cells, but no inhibitory effect on the growth of HepG2 and L929 cells (F=13112.13, P=0.0001). In the chick choriallantic membrane (CAM) assay, the expressed fusion protein significantly inhibited neovascularization. Rabbit inflammatory corneal neovascularization (CNV) induced by intrastromal sutures resulted in a uniform neovascular response. In this model, direct subconjunctival injection of AdhENDO VEGI151 expressed the fusion protein in vivo and suppressed the development of CNV. Topical application of AdhENDO-VEGI151 led to a significant suppression of CNV (F=1413.11, P=0.0001), as compared with the control group of AdLacZ. Immunohistochemical staining showed the fusion protein dominantly expressed in corneal epithelium. Compared with the control group of AdLacZ (4075.9+/-1849.9 mm(3)), the average tumor size of group AdhENDO-VEGI151 reduced in size (487.7+/ 241.2 mm(3)) (F=14.80, P=0.0085), with an inhibition rate of 88.03%. Immunohistochemical staining showed the adenoviruses carried the fusion gene expressed on liver cancer cell membrane. MVD decreased more significantly in treated mice (30.75+/-3.31%) than in AdLacZ control (50.25+/-8.65%) (F=17.72, P=0.0056) with an inhibition rate of 39%. CONCLUSION: Fusion protein expressed by recombinant adenoviruses has a significant inhibitory effect on neovascularization. PMID- 15133845 TI - Changes of tumor microcirculation after transcatheter arterial chemoembolization: first pass perfusion MR imaging and Chinese ink casting in a rabbit model. AB - AIM: To observe the change of tumor microcirculation after transcatheter arterial chemoembolization (TACE) with bletilla microspheres by using first pass perfusion MR imaging (FP) and Chinese ink casting. METHODS: VX2 carcinoma cells were surgically implanted into the left and right lobes of liver of 30 New Zealand white rabbits, which were divided into 3 groups at random. Emulsion of lipiodol mixed with mitomycin C, and 5-FU bletilla microspheres were injected into the hepatic artery respectively, and saline was used as control agent. MR imaging was performed with turbo-flash sequence 14 d after tumor implantation and 7 d after interventional therapy. The steepest slopes (SS) of the signal intensity versus time curves were created for quantitative analysis, 7.5% Chinese ink gelatin solution was injected through ascending artery (17 cases) or portal vein (2 cases) for lesion microvessel area (MVA) measurement after the last MRI examination. The correlation between perfusion imaging and MVA was studied blindly. RESULTS: The SS values at the rim of tumor in lipiodol group (mean, 49% per second) and bletilla group (mean, 35% per second) were significantly decreased (P<0.05) as compared with control group (mean, 124% per second), no difference was found between lipiodol and bletilla groups (P>0.05). In lipiodol group, the MVAs (24 974+/-11 836 microm(2)) in the center of the tumor were significantly smaller than those of the control group (35 510+/-15 675 microm(2)) (P<0.05), while the MVAs (80 031+/-22 745 microm(2)) around the tumor were significantly increased because small and dense plexuses appeared around the tumor which correlated to intense reaction of granulation tissue. None of the vessels was seen in the tumor in bletilla group, the peripheral MVAs of the tumor were significantly smaller than those of the control group (P<0.05) and lipiodol group (P<0.05). There was a good correlation between SS and MVAs in control group (r(s), 0.985, P<0.0001) and bletilla group (r(s), 0.743, P<0.05), the correlation was not significant in lipiodol group (r(s), 0.527, P>0.05). CONCLUSION: TACE with bletilla microspheres may enhance its anti-tumor effect by inhibiting the angiogenesis, and FP-MRI provides useful information to assess the TACE effect by depicting tumor vascularization and perfusion. PMID- 15133846 TI - Expression of angiostatin cDNA in human hepatocellular carcinoma cell line SMMC 7721 and its effect on implanted carcinoma in nude mice. AB - AIM: To transfect murine angiostatin cDNA into human hepatocellular carcinoma cell line SMMC-7721 and to investigate its effects on implanted carcinoma in nude mice. METHODS: A eukaryotic expression vector of pcDNA3.1-mAST containing murine angiostatin was constructed. Then pcDNA3.1-mAST plasmid was transfected into cell line SMMC-7721 by Lipofectamine. The resistant clone was screened by G418 filtration and identified by RT-PCR and Western blotting. Nude mice were divided into three groups of 10 each. Mice in blank control group were only injected with SMMC-7721 cells. Mice in vector control group were injected with SMMC-7721 cells transfected with pcDNA3.1 (+) vector, whereas mice in angiostatin group were injected with SMMC-7721 cells transfected with pcDNA3.1-mAST plasmid. Volume, mass and microvessel density (MVD) of the tumors in different groups were measured and compared. RESULTS: Murine angiostatin cDNA was successfully cloned into the eukaryotic expression vector pcDNA3.1 (+). pcDNA3.1-mAST was successfully transfected into SMMC-7721 cell line and showed stable expression in this cell line. No significant difference was observed in the growth speed of SMMC-7721 cells between groups transfected with and without angiostatin cDNA. Tumor volume, mass and MVD in the angiostatin group were significantly lower than those in the blank control group and vector control group (P<0.01). The inhibitory rate of tumor reached 78.6%. Mass and MVD of the tumors only accounted for 34.6% and 48.9% respectively of those in the blank control group. CONCLUSION: Angiostatin cDNA could be stably expressed in human hepatocellular carcinoma cell line SMMC-7721 without obvious inhibitory effects on the growth of SMMC-7721 cells. When implanted into nude mice, SMMC-7721 cells transfected with angiostatin cDNA show a decreased tumorigenic capability. It suggests that angiostatin can inhibit tumor growth through its inhibition on angiogenesis in tumors. PMID- 15133847 TI - Expression and altered subcellular localization of the cyclin-dependent kinase inhibitor p27Kip1 in hepatocellular carcinoma. AB - AIM: To investigate p27 expression in hepatocellular carcinoma (HCC), adjacent nontumoral and normal liver tissues, and to verify whether the subcellular localization of p27 was altered in HCC. METHODS: The level of p27 in tumoral, nontumoral, and normal liver tissues were assessed by immunohistochemical (IHC) analysis. Parallel immunostaining was done for proliferating cell nuclear antigen (PCNA) to evaluate cell proliferation. RESULTS: The labeling index (LI) of p27 in tumoral lesions was significantly lower than that in adjacent nontumoral lesions (t=2.444, P=0.017) and normal controls (t=2.268, P=0.029). The LI of p27 significantly decreased in patients with massive type (t=2.227, P=0.037) and infiltration (t=2.197, P=0.036). The prognosis of patients with higher p27 LI was longer than that of patients with lower p27 LI (P=0.0247, log-rank test). The LI of PCNA was significantly higher in HCC than that in adjacent nontumoral lesions (t=2.092, P=0.041) and normal controls (t=3.533, P=0.002). There was no significant correlation between p27 expression and cell proliferation in tumor samples. The level of p27 in the cytoplasmic fraction was higher in tumoral and nontumoral liver tissues, and was associated with clinical stage (t=2.520, P=0.029) and the degree of invasion (t=2.640, P=0.019). Survival analysis showed that p27 was an independent prognosis marker for HCC patients. CONCLUSION: These results suggest that p27 underexpressing in patients with HCC is closely associated with infiltration, metastasis, and prognosis. Alterations in the subcellular localization of p27 protein may occur early during hepatocarcinogenesis. PMID- 15133848 TI - Loss of heterozygosity on chromosome 1 in sporadic colorectal carcinoma. AB - AIM: Loss of heterozygosity (LOH) on tumor suppressor genes is believed to play a key role in carcinogenesis of colorectal cancer. When it occurs at a tumor suppressor gene locus with abnormal allele, neoplastic transformation happens. In this study, we analyzed the LOH at 21 loci on chromosome 1 in sporadic colorectal cancer to identify additional loci involved in colorectal tumorigenesis. METHODS: Twenty-one polymorphic micro-satellite DNA markers were analyzed with PCR both in 83 cases of colorectal cancer and in normal tissues. PCR products were eletrophoresed on an ABI 377 DNA sequencer. Genescan 3.1 and Genotype 2.1 software were used for LOH scanning and analysis. chi2 test was used to compare LOH frequency with clinicopathological data. P<0.05 was considered as statistically significant. RESULTS: The average LOH frequency of chromosome 1, short arm and long arm was 19.83%, 18.00% and 21.66%, respectively. The 2 highest LOH loci with a frequency of 36.54% and 32.50% were identified on D1S468 (1p36.33 p36.31) and D1S413 (1q31.3), respectively. On D1S2726 locus, LOH frequency of rectal cancer was 28.57% (6/21), which was higher than that of colon cancer (0.00%, 0/33) (P=0.002), suggesting that the mechanism of carcinogenesis was different in both groups. CONCLUSION: Putative tumor suppressor genes on chromosome 1 may relate to sporadic colorectal carcinomas. Tumor-suppressor-genes might locate on 1p36.33-36.31 and/or 1q31.3. PMID- 15133849 TI - Effect of Hejie decoction on T cell immune state of chronic hepatitis B patients. AB - AIM: To explore the effect of Hejie decoction (HJD) (mediation decoction) on T cellular immune state of chronic hepatitis B patients. METHODS: Sixty-five patients with chronic hepatitis B were randomly divided into 2 groups. Forty patients in the treatment group were treated by HJD, and 25 patients in the control group were treated by routine Western medicine. The TCRVbeta7 gene expression, T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) levels were observed before and after treatment. RESULTS: The level of CD4+ cells was lower whereas the level of CD8+ cells was higher in patients than in the normal group. There was no significant difference between the levels of CD3+ cells in patients and normal persons. After 6 months of treatment, ALT, AST, TB levels of the 2 groups were obviously decreased, and the level of CD4+ cells was increased whereas the level of CD8+ cells was decreased in the treatment group. However, the level of CD4+ cells and CD8+ cells had no significant difference in the control group. TCRVbeta7 expressions were detected in 6 patients of the treatment group, whose HBV-DNA and HBeAg turned negative and ALT became normal. HBeAg in another 3 patients turned negative while HBV-DNA did not, and TCRVbeta7 expressions were not detectable. TCRVbeta7 expression could not be detected in the control group, HBV-DNA of the control group did not turn negative. HBeAg in 1 patient turned negative while HBV-DNA did not, and TCRVbeta7 expressions were not detectable. The total effective rate was not significantly different between the 2 groups and the markedly effective rate was significantly different (P<0.01). CONCLUSION: HJD is effective for treating chronic hepatitis B, and its effect seems to relate with the improvement of the TCRVbeta7 expression of chronic hepatitis B patients, thus activating T cells and eliminating HBV. T cellular immune function plays an important role in HBV infection and virus elimination. PMID- 15133850 TI - Differential expression of cholangiocyte and ileal bile acid transporters following bile acid supplementation and depletion. AB - AIM: We have previously demonstrated that cholangiocytes, the epithelial cells lining intrahepatic bile ducts, encode two functional bile acid transporters via alternative splicing of a single gene to facilitate bile acid vectorial transport. Cholangiocytes possess ASBT, an apical sodium-dependent bile acid transporter to take up bile acids, and t-ASBT, a basolateral alternatively spliced and truncated form of ASBT to efflux bile acids. Though hepatocyte and ileal bile acid transporters are in part regulated by the flux of bile acids, the effect of alterations in bile acid flux on the expression of t-ASBT in terminal ileocytes remains unclear. Thus, we tested the hypothesis that expression of ASBT and t-ASBT in cholangiocytes and ileocytes was regulated by bile acid flux. METHODS: Expression of ASBT and t-ASBT message and protein in cholangiocytes and ileocytes isolated from pair-fed rats given control (C) and 1% taurocholate (TCA) or 5% cholestyramine (CY) enriched diets, were assessed by both quantitative RNase protection assays and quantitative immunoblotting. The data obtained from each of the control groups were pooled to reflect the changes observed following TCA and CY treatments with respect to the control diets. Cholangiocyte taurocholate uptake was determined using a novel microperfusion technique on intrahepatic bile duct units (IBDUs) derived from C, TCA and CY fed rats. RESULTS: In cholangiocytes, both ASBT and t-ASBT message RNA and protein were significantly decreased in response to TCA feeding compared to C diet. In contrast, message and protein of both bile acid transporters significantly increased following CY feeding compared to C diet. In the ileum, TCA feeding significantly up-regulated both ASBT and t-ASBT message and protein compared to C diet, while CY feeding significantly down-regulated message and protein of both bile acid transporters compared to C diet. As anticipated from alterations in cholangiocyte ASBT expression, the uptake of taurocholate in microperfused IBDUs derived from rats on TCA diet decreased 2.7-fold, whereas it increased 1.7-fold in those on CY diet compared to C diet fed groups. CONCLUSION: These data demonstrate that expression of ASBT and t-ASBT in cholangiocytes is regulated by a negative feedback loop while the expression of these transporters in terminal ileum is modified via positive feedback. Thus, while transcriptional regulatory mechanisms in response to alterations in bile acid pool size are operative in both cholangiocytes and ileocytes, each cell type responds differently to bile acid supplementation and depletion. PMID- 15133851 TI - Transcriptional regulation of human alpha1(I) procollagen gene in dermal fibroblasts. AB - AIM: To clarify the fractional activity of promoters from human alpha1(I) procollagen gene, the interaction between cis-elements and consensus DNA-binding proteins responsible for high promoter activity, and the potential application of promoter competitors as well as cytokines for antifibrogenesis. METHODS: Sequence between 2483 bp upstream of the start of transcription and 42 bp downstream of this site was investigated with serial 5'-deletion. The 5'-deleted promoters recombined with chloramphenicol acetyltransferase (CAT) as reporter gene were transiently transfected to human dermal fibroblasts. Electrophoretic mobility shift assay was performed to show the DNA-protein binding capacity of the promoter sequence. Cytokines including tumor necrosis factor alpha (TNFalpha) and interferons (INFs) were added to the culture medium of transiently transfected fibroblasts. Competitor DNA for the binding sites of Sp-1, Ap-1 and NF-1 was individually cotransfected transiently in order to block the promoter-driven CAT expression. RESULTS: Sequences of -2483 to +42 bp and -268 to +42 bp of human alpha1(I) procollagen gene had high activity as promoters. Binding sites for Ap-1 and Sp-1 were among the cis-regulatory elements recognizing consensus transcription factors responsible for basal promoter activity of sequence -268 to +42 bp. TNFalpha, IFNalpha, IFNbeta showed inhibitory effects on sequence -2 483 to +42 bp as promoter with activities 43%, 62% and 60% of control respectively. Transfection of the promoter competitors could reverse the promoter activity of 268 to +42 bp 40-60%. CONCLUSION: Sequences of -2 483 to +42 bp recombined with reporter gene provide an ideal construction for transcriptional study of alpha1(I) procollagen gene. The anti-collagen capacity of TNFalpha and IFNs is associated with their transcriptional regulation. Ap-1 and Sp-1 mediate the basal transcriptional activation of human alpha1(I) procollagen gene in dermal fibroblasts. Competitors for highly active promoters might be a novel potential candidate in fibrotic blockade. PMID- 15133852 TI - In vitro cultivation of human fetal pancreatic ductal stem cells and their differentiation into insulin-producing cells. AB - AIM: To isolate, culture and identify the human fetal pancreatic ductal stem cells in vitro, and to observe the potency of these multipotential cells differentiation into insulin-producing cells. METHODS: The human fetal pancreas was digested by 1 g/L collagease type IV and then 2.5 g/L trypsin was used to isolate the pancreatic ductal stem cells, followed by culture in serum-free, glucose-free DMEM media with some additional chemical substrates in vitro (according to the different stage). The cells were induced by glucose-free (control), 5 mmol/L, 17.8 mmol/L and 25 mmol/L glucose, respectively. The cell types of differentiated cells were identified using immunocytochemical staining. RESULTS: The shape of human fetal pancreatic ductal stem cells cultured in vitro was firstly fusiform in the first 2 wk, and became monolayer and cobblestone pattern after another 3 to 4 wk. After induced and differentiated by the glucose of different concentrations for another 1 to 2 wk, the cells formed the pancreatic islet-like structures. The identification and potency of these cells were then identified by using the pancreatic ductal stem cell marker, cytokeratin 19 (CK-19), pancreatic beta cell marker, insulin and pancreatic alpha cell marker, glucagons with immunocytochemical staining. At the end of the second week, 95.2% of the cells were positive for CK-19 immunoreactivity. Up to 22.7% of the cells induced by glucose were positive for insulin immunoreactivity, and less than 3.8% of the cells were positive for glucagon immunoreactivity in pancreatic islet-like structures. The positive ratio of immunoreactive staining was dependent on the concentration of glucose, and it was observed that the 17.8 mmol/L glucose stimulated effectively to produce insulin- and glucagons-producing cells. CONCLUSION: The human fetal pancreatic ductal stem cells are capable of proliferation in vitro. These cells have multidifferentiation potential and can be induced by glucose and differentiated into insulin-producing cells in vitro. PMID- 15133853 TI - Immune tolerance in pancreatic islet xenotransplantation. AB - AIM: To observe the effect of tail vein injection with donor hepatocytes and/or splenocytes on the islet xenotransplantation rejection. METHODS: New-born male pigs and BALB/C mice were selected as donors and recipients respectively. Islet xenotransplantation was performed in recipients just after the third time of tail vein injection with donor hepatocytes and/or splenocytes. Macrophage phagocytosis, NK(natural killing cell) killing activity, T lymphocyte transforming function of spleen cells, antibody forming function of B lymphocytes, and T lymphocyte subsets were taken to monitor transplantation rejection. The effects of this kind of transplantation were indicated as variation of blood glucose and survival days of recipients. RESULTS: The results showed that streptozotocin (STZ) could induce diabetes mellitus models of mice. The pre-injection of donor hepatocytes, splenocytes or their mixture by tail vein injection was effective in preventing donor islet transplantation from rejection, which was demonstrated by the above-mentioned immunological marks. Each group of transplantation could decrease blood glucose in recipients and increase survival days. Pre-injection of mixture of donor hepatocytes and splenocytes was more effective in preventing rejection as compared with that of donor hepatocyte or splenocyte pre-injection respectively. CONCLUSION: Pre-injection of donor hepatocytes, splenocytes or their mixture before donor islet transplantation is a good way in preventing rejection. PMID- 15133854 TI - Establishment and characterization of human hepatocellular carcinoma cell line FHCC-98. AB - AIM: To establish a novel human hepatocellular carcinoma (HCC) cell line FHCC-98 from HCC tissue and to provide a suitable model for studying HCC occurrence, progress and metastasis. METHODS: Serially passaged cells were cultured and their morphologies were observed under light and electron microscope. Cytogenetic study was conducted by using flow cytometry and chromosome analysis. Expressions of tumor markers such as alpha-fetoprotein (AFP), cytokeratin (CK) and hepatoma metastasis-associated factor HAb18G/CD147 on the FHCC-98 cells were detected by immunocytochemistry or Western blotting. Lactic dehydrogenase (LDH) isoenzymes were detected by polyacrylamide gel electrophoresis (PAGE). Xenograft was performed by inoculating FHCC-98 cells into the flanks of nude mice. RESULTS: Morphology of FHCC-98 cells was the same as that of other malignant cells. The expressions of the cells were positive for HAb18G/CD147 and CK, and negative for AFP. Its population doubling time was 21.4 h. The cell DNA was tetraploid and the major chromosomes were triploid by cytogenetics analysis. The tumorigenicity in nude mice was 100%. PAGE showed four bands representing LDH2, LDH3, LDH4 and LDH5. CONCLUSION: FHCC-98 is a novel HCC cell line and an ideal cell model for further exploring the mechanism of hepatocellular carcinoma invasion and metastasis. PMID- 15133856 TI - Mesenteric artery remodeling and effects of imidapril and irbesartan on it in spontaneously hypertensive rats. AB - AIM: To investigate the remodeling of mesenteric artery and the expression of TGF beta1, c-Jun in mesenteric artery and effects of imidapril and irbesartan on the remodeling in spontaneously hypertensive rats (SHR). METHODS: Thirty SHR (male/female, 21/9), aged 13 wk, were randomly divided into 3 groups (7 male rats and 3 female rats each group): SHR group, imidapril group (imidapril 3 mg/kg.d was given in drinking water for 14 wk), and irbesartan group (irbesartan 50 mg/kg.d was given in drinking water foe 14 wk). Ten homogeneous Wistar Kyoto rats, 5 males and 5 females, weighing 206+/-49 g, were selected as normal control group (WKY group). Systolic pressure was measured on d 1, 2, 4, 6, 8, 10, 12 and 14 during the experiment and the rats were killed at the end of the experiment. Angiotensin II (Ang II) level in plasma and mesenteric arteries was measured by radioimmunoassay. The morphology of the secondary branches of mesenteric artery were examined by light microscopy and electron microscopy. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of transforming growth factor TGF-beta1 and c-Jun mRNA. RESULTS: Compared with imidapril group and irbesartan group, the blood pressure was remarkably increased in SHR group. Ang II level in plasma and mesenteric arteries in SHR group was the same or lower than that in WKY group, and was higher in irbesartan group and lower in imidapril group. The remodeling of mesenteric arteries in SHR group was mostly obvious among the 4 groups. The ratio of TGF-beta1 absorbed light value to GAPDH absorbed light value in the SHR group was 0.887+/-0.019, which was significantly higher than that in WKY group, imidapril group, and irbesartan group with the ratios of 0.780+/-0.018, 0.803+/-0.005, and 0.847+/-0.017, respectively (P<0.01). Ang II level in plasma and mesenteric arteries in imidapril group was significantly lower than that in irbesartan group (P<0.05). The c-Jun absorbed light value/GAPDH absorbed light value of mesenteric arteries in the SHR group was 0.850+/-0.015, which was significantly higher than that in the WKY, imidapril, and irbesartan groups (0.582+/-0.013, 0.743+/-0.012, and 0.789+/-0.013, respectively, P<0.01), and was significantly lower in imidapril group than in irbesartan group (P<0.05). CONCLUSION: Imidapril and irbesartan can not only control blood pressure but also inhibit mesenteric arteries remodeling and mRNA expression of TGF-beta1, c-Jun in SHR. Imidapril is more effective than irbesartan. PMID- 15133855 TI - Overexpression of annexin 1 in pancreatic cancer and its clinical significance. AB - AIM: To investigate the expression of annexin I in pancreatic cancer and its relationship with the clinicopathologic factors, and to evaluate its potential clinical significance. METHODS: Annexin I expression was analyzed by Western blot and immunohistochemical staining in pancreatic adenocarcinoma and multi-tissue microarrays (MTAs). RESULTS: Western blot analysis showed that annexin I was overexpressed in 84.6% (11/13) pancreatic ductal adenocarcinomas. Immunohistochemistry analysis of pancreatic cancer in MTAs showed that annexin I protein was 71.4%(30/42) positive which was markedly increased compared with that in the tumor matched normal pancreas tissues 18.4%(7/38) (P<0.01). In the meantime, the high expression of annexin 1 was correlated with the poor differentiation of pancreatic adenocarcinoma. CONCLUSION: Annexin 1 overexpression is a frequent biological marker and correlates with the differentiation of pancreatic cancer during tumorigenesis. PMID- 15133857 TI - Signal pathways involved in emodin-induced contraction of smooth muscle cells from rat colon. AB - AIM: To investigate the effects induced by emodin on single smooth muscle cells from rat colon in vitro, and to determine the signal pathways involved. METHODS: Cells were isolated from the muscle layers of Wistar rat colon by enzymatic digestion. Cell length was measured by computerized image micrometry. Intracellular Ca2+ ([Ca2+]i) signals were studied using the fluorescent Ca2+ indicator fluo-3 and confocal microscopy. PKCalpha distribution at rest state or after stimulation was measured with immunofluorescence confocal microscopy. RESULTS: (1) Emodin dose-dependently caused colonic smooth muscle cells contraction; (2) emodin induced an increase in intracellular Ca2+ concentration; (3) the contractile responses induced by emodin were respectively inhibited by preincubation of the cells with ML-7 (an inhibitor of MLCK) and calphostin C (an inhibitor of PKC); (4) Incubation of cells with emodin caused translocation of PKCalpha from cytosolic area to the membrane. CONCLUSION: Emodin has a direct contractile effect on colonic smooth muscle cell. This signal cascade induced by emodin is initiated by increased [Ca2+]i and PKCalpha translocation, which in turn lead to the activation of MLCK and the suppression of MLCP. Both of them contribute to the emodin-induced contraction. PMID- 15133858 TI - Characterization and enrichment of hepatic progenitor cells in adult rat liver. AB - AIM: To detect the markers of oval cells in adult rat liver and to enrich them for further analysis of characterization in vitro. METHODS: Rat model for hepatic oval cell proliferation was established with 2-acetylaminofluorene and two third partial hepatectomy (2-AAF/PH). Paraffin embedded rat liver sections from model (11 d after hepatectomy) and control groups were stained with HE and OV6, cytokeratin19 (CK19), albumin, alpha fetoprotein (AFP), connexin43, and c-kit antibodies by immunohistochemistry. Oval cell proliferation was measured with BrdU incorporation test. C-kit positive oval cells were enriched by using magnetic activated cell sorting (MACS). The sorted oval cells were cultured in a low density to observe colony formation and to examine their characterization in vitro by immunocytochemistry and RT-PCR. RESULTS: A 2-AAF/PH model was successfully established to activate the oval cell compartment in rat liver. BrdU incorporation test of oval cell was positive. The hepatic oval cells coexpressed oval cell specific marker OV6, hepatocyte-marker albumin and cholangiocyte-marker CK19. They also expressed AFP and connexin 43. C-kit, one hematopoietic stem cell receptor, was expressed in hepatic oval cells at high levels. By using c-kit antibody in conjunction with MACS, we developed a rapid oval cell isolation protocol. The sorted cells formed colony when cultured in vitro. Cells in the colony expressed albumin or CK19 or coexpressed both and BrdU incorporation test was positive. RT-PCR on colony showed expression of albumin and CK19 gene. CONCLUSION: Hepatic oval cells in the 2-AAF/PH model had the properties of hepatic stem/progenitor cells. Using MACS, we established a method to isolate oval cells. The sorted hepatic oval cells can form colony in vitro which expresses different combinations of phenotypic markers and genes from both hepatocytes and cholangiocyte lineage. PMID- 15133860 TI - Synthesis of ribozyme against vascular endothelial growth factor165 and its biological activity in vitro. AB - AIM: To investigate the designation, synthesis and biological activity of against vascular endothelial growth factor165 (VEGF165) ribozyme. METHODS: The ribozyme against VEGF165 was designed with computer. The transcriptional vector was constructed which included the anti-VEGF165 ribozyme and 5', 3' self-splicing ribozymes. The hammerhead ribozyme and substrate VEGF165 mRNA were synthesized through transcription in vitro. The cleavage activity of the ribozyme on target RNA was observed in a cell-free system. RESULTS: The anti-VEGF165 ribozyme was released properly from the transcription of pGEMRz212 cleaved by 5' and 3' self splicing ribozymes which retained its catalytic activity, and the cleavage efficiency of ribozyme reached 90.7%. CONCLUSION: The anti-VEGF165 ribozyme designed with computer can cleave VEGF165 mRNA effectively. PMID- 15133859 TI - Influence of serum collected from rat perfused with compound Biejiaruangan drug on hepatic stellate cells. AB - AIM: To observe the effect of compound Biejiaruangan decoction (CBJRGC) (composite prescription of Carapax trionycis for softening the liver) on proliferation, activation, excretion of collagen and cytokine of hepatic stellate cells (HSCs) and to find the mechanism of prevention and treatment of hepatic fibrosis by CBJRGC. METHODS: Using MTT, immunohistochemistry and image analysis technology, the related indexes for proliferation, activation, excretion of collagen and cytokine of hepatic stellate cells were detected in 24 h, 48 h, and 72 h after administration of different dosages of CBJRGC. RESULTS: Statistical analysis showed that serum collected from rat perfused with CBJRGC could restrain the proliferation of HSC in 48 h and 72 h especially in high and medium dosage groups, markedly decrease the expression of desmin, synapsin and platelet derived growth factor (PDGF) in HSC in 24 h, 48 h and 72 h, as well as the expression of alpha-SMA, collagen III, TIMP and TGFbeta1 in 48 h and 72 h, decrease the excretion of collagen I in 72 h. CBJRGC serum had no significant effect on collagens I, III and TIMP in 24 h. CONCLUSION: CBJRGC serum has a good curative effect on hepatic fibrosis. Its main mechanism may be related to the following factors. The drug serum can restrain the proliferation and activation of HSC, decrease the number of activated HSC and the total number of HSC, the excretion of collagens I, III, enhance the degradation of collagen and restore the balance of synthesis and degradation of collagen, inhibit the expression of transforming growth factor beta1 (TGFalpha1) and platelet derived growth factor (PDGF) in HSC, block and delay the process of hepatic fibrosis. Synapsin is a new marker of activation of HSC, which provides a theoretical and testing basis for neural regulation in the developing process of hepatic fibrosis. PMID- 15133861 TI - Auxiliary en-bloc liver-small bowel transplantation with partial pancreas preservation in pigs. AB - AIM: The aim of this study was to describe an auxiliary combined liver-small bowel transplantation model with the preservation of duodenum, head of pancreas and hepatic biliary system in pigs. The technique, feasibility, security and immunosuppression were commented. METHODS: Forty outbred long-white pigs were randomized into two groups, and the auxiliary composite liver/small bowel allotransplantations were undertaken in 10 long-white pigs in each group with the recipient liver preserved. Group A was not treated with immunosuppressive drugs while group B was treated with cyclosporine A and methylprednisolone after operation. The hemodynamic changes and amylase of body fluid (including blood, urine and abdominal drain) were analyzed. RESULTS: The average survival time of the animals was 10+/-1.929 d (6 to 25 d) in group A while more than 30 d in group B. The pigs could tolerate the hemodynamic fluctuation during operation and the hemodynamic parameters recovered to normal 2 h after blood reperfusion. The transient high amylase level was decreased to normal one week after operation and autopsy showed no pancreatitis. CONCLUSION: Auxiliary en-bloc liver-small bowel transplantation with partial pancreas preservation is a feasible and safe model with simplified surgical techniques for composite liver/small bowel transplantation. This model may be used as a preclinical training model for clinical transplantation method, clinical liver-small bowel transplantation related complication research, basic research including immunosuppressive treatment, organ preservation, acute rejection, chronic rejection, immuno tolerance and xenotransplantation. PMID- 15133862 TI - Biochemical and radiological predictors of malignant biliary strictures. AB - AIM: Differentiation of benign biliary strictures (BBS) from malignant biliary strictures (MBS) remains difficult despite improvement in imaging and endoscopic techniques. The aim of this study was to identify the clinical, biochemical and or radiological predictors of malignant biliary strictures. METHODS: We retrospectively reviewed all charts of patients who had biliary strictures (BS) on endoscopic retrograde cholangiopancreatography (ERCP) or percutaneous cholangiography (PTC) in case of unsuccessful ERCP from March 1998 to August 2002. Patient characteristics, clinical features, biochemical, radiological and biopsy results were all recorded. Stricture etiology was determined based on cytology, biopsy or clinical follow-up. A receiver operator characteristic (ROC) curve was constructed to determine the optimal laboratory diagnostic criterion threshold in predicting MBS. RESULTS: One hundred twenty six patients with biliary strictures were enrolled, of which 72 were malignant. The mean age for BBS was 53 years compared to 62.4 years for MBS (P=0.0006). Distal bile duct stricture was mainly due to a malignant process 48.6% vs 9% (P=0.001). Alkaline phosphates and AST levels were more significantly elevated in MBS (P=0.0002). ROC curve showed that a bilirubin level of 84 micromol/L or more was the most predictive of MBS with a sensitivity of 98.6%, specificity of 59.3% and a positive likelihood ratio of 2.42 (95% CI=0.649-0.810). Proximal biliary dilatation was more frequently encountered in MBS compared to BBS, 73.8% vs 39.5% (P=0.0001). Majority of BBS (87%) and MBS (78%) were managed endoscopically. CONCLUSION: A serum bilirubin level of 84 micromol/L or greater is the best predictor of MBS. Older age, proximal biliary dilatation, higher levels of bilirubin, alkaline phosphatase, ALT and AST are all associated with MBS. ERCP is necessary to diagnose and treat benign and malignant biliary strictures. PMID- 15133863 TI - Polymorphisms at cholesterol 7alpha-hydroxylase, apolipoproteins B and E and low density lipoprotein receptor genes in patients with gallbladder stone disease. AB - AIM: To investigate the relationship between gallbladder stone disease (GSD) and single nucleotide polymorphisms of cholesterol 7alpha-hydroxylase (CYP7A) gene promoter, apolipoprotein (APO) B gene exon 26, APOE gene exon 4 or microsatellite polymorphism of low density lipoprotein receptor (LDLR) gene exon 18. METHODS: Genotypes of CYP7A, APOB, APOE and LDLR genes were determined in 105 patients with GSD diagnosed by B-mode ultrasonography and 274 control subjects. Serum lipids were analyzed with HITACHI 7060 automatic biochemical analyzer. RESULTS: Body mass index (BMI) was significantly higher in patients with GSD (24.47+/ 3.09) than in controls (23.50+/-2.16). Plasma total cholesterol was lower in patients with GSD (4.66+/-0.92 mmol/L) than in controls (4.91+/-0.96 mmol/L), P<0.01 after adjusted for age, sex and BMI. The significantly higher frequency of A allele of CYP7A gene polymorphism and X+ allele of APOB gene polymorphism was seen in GSD patients. Percentages of A allele in patients and controls were 62.86% and 54.38% (P<0.05) and those of X+ allele 8.57% and 4.01% (P<0.01). Subjects with A allele had significantly lower plasma total cholesterol and LDL cholesterol than subjects with CC homozygote. In a multiple variable logistic regression model, the BMI (OR=1.13, 95% CI: 1.05-1.22), A allele (OR=1.48, 95% CI: 1.05-2.09) and X+ allele (OR=2.28, 95% CI: 1.14-4.59) were positively associated with GSD (P<0.05). Plasma total cholesterol (OR=0.69, 95% CI: 0.64 0.74) was negatively related to GSD (P<0.05). CONCLUSION: With an association analysis, it was determined that A allele of CYP7A gene and X+ allele of APOB gene might be considered as risk genes for GSD. These alleles are related with differences of serum lipids among subjects. Multiple-variable logistic regression model analysis showed that besides BMI, GSD was affected by polygenetic factors. But the mechanism for these two alleles responsible for GSD requires further investigations. PMID- 15133864 TI - Different therapy for different types of ulcerative colitis in China. AB - AIM: To study the different therapy for different types of ulcerative colitis (UC) in China. METHODS: Among 102 UC patients, 42 chronic relapse type UC patients were randomly divided into olsalazine sodium treatment group (n=21) and SASP group (n=21). Clinical effects and safety were observed in the 2 groups. Forty-two first episode type UC patients were randomly divided into Heartleaf houttuynia herb treatment group (n=21) and SASP group (n=21). Clinical effects were observed in the 2 groups while ultrastructure of colonic mucosa, ICAM-1 and the pressure of distant colon were studied in Heartleaf houttuynia herb group. Eighteen patients (8 males, 10 females) with refractory UC and unresponsive to high-dose prednisolone and sulfasalazine therapy more than one month were treated with Kangshuanling (7200 U/d). Prednisolone was gradually stopped and sulfasalazine was maintained. Stool frequency, rectal bleeding, colonoscopy, general well-being, histology were observed and CD62p, CD63, CD54, Pgp-170 (flow cytometry), TXA2 (RIA), blood platelet aggregation rate and thrombosis length in vitro were assessed. RESULTS: In the 42 chronic relapse type UC patients, the overall clinical effects of olsalazine sodium group (complete remission in 16, improvement in 4, inefficiency in 1) were better than those of SASP group (complete remission in 10, improvement in 4, inefficiency in 7, P<0.05). Symptomatic remission of olsalazine sodium group (complete remission in 15, partial remission in 5, inefficiency in 1) was better than that of SASP group (complete remission in 10, partial remission in 5, inefficiency in 6, P<0.05). The colonoscopic remission of olsalazine sodium group(complete remission in 11, partial remission in 9, inefficiency in 1) was better than that of SASP group (complete remission in 7, partial remission in 8, inefficiency in 6, P<0.05). The histologic remission of olsalazine sodium group (complete remission in 13, partial remission in 7, inefficiency in in 1) was better than that of SASP group (complete remission in 6, partial remission in 10, inefficiency in 5, P<0.05). The side effects of gastrointestinal tract in olsalazine sodium group were less than those of SASP group except for frequency of watery diarrhea. No other side effects were observed in olsalazine sodium group while ALT increase, WBC decrease and skin eruption were observed in SASP group. Two patients relapsed in olsalazine sodium group while 8 cases relapsed in SASP group during the flow-up period (from six months to one year). In the 42 first episode type UC patients, the clinical effect of Heartleaf houttuynia herb group (complete remission in 20, 95.2%; improvement in 1, 4.8%) was better than that of SASP group (complete remission in 15, 72.4%, improvement in 5, 23.8%; inefficiency in 1, 3.8%, P<0.01). The time of stool frequency recovering to normal (5.6+/-3.3 d), and blood stool disappearance (6.7+/-3.8 d) and abdominal pain disappearance (6.1+/ 3.5 d) in Heartleaf houttuynia herb group was all shorter than that in SASP group (9.5+/-4.9 d, 11.7+/-6.1 d, 10.6+/-5.3 d, P<0.01). Heartleaf houttuynia herb could inhibit the epithelial cell apoptosis of colonic mucous membrane and the expression of ICAM-1 (45.8+/-5.7% vs 30.7+/-4.1%, P<0.05). Compared with normal persons, the mean promotive speed of contraction wave stepped up (4.6+/-1.6 cm/min vs 3.2+/-1.8 cm/min, P<0.05) and the mean amplitude of the wave decreased (14.2+/-9.3 kPa vs 18.4+/-8.0 kPa, P<0.05) in active UC patients. After treatment with Heartleaf houttuynia herb, these 2 indexes improved significantly (17.3+/ 8.3 kPa, 3.7+/-1.7 cm/min, P<0.05). In normal persons, the postprandial pressure of sigmoid (2.9 +/-0.9 kPa) was higher than that of descending colon (2.0+/-0.7 kPa) and splenic flexure (1.7+/-0.6 kPa), while the colonic pressure (1.5+/-0.5 kPa, 1.4+/-0.6 kPa, 1.3+/-0.6 kPa) decreased significantly (P<0.05) in active UC patients. After treatment with Heartleaf houttuynia herb, the colonic pressure (2.6+/-0.8 kPa, 1.8+/-0.6 kPa, 1.6+/-0.5 kPa) recovered to normal. The pain threshold Heartleaf houttuynia herb, the colonic pressure (2.6+/-0.8 kPa, 1.8+/ 0.6 kPa, 1.6+/-0.5 kPa) recovered to normal. The pain threshold of distant colon (67.3+/-18.9 mL) in active UC patients decreased significantly compared with that of normal persons (216.2+/-40.8 mL, P<0.05) and recovered to normal after treatment with Heartleaf houttuynia herb(187.4+/-27.2 mL, P<0.05). In the 18 refractory UC patients with platelet activation, after more than 4 wk of combined Kangshuanling and sulfasalazine therapy, 16 patients achieved clinical remission, with a highly significant statistical difference (P<0.01) between pre-and post treatment mean scores for all disease parameters: stool frequency (8.2/d vs 1.6/d), rectal bleeding (score 2.7 vs 0.3), colonoscopy (score 2.6 vs 1.1), histology (score 12.0 vs 5.0), general well being (score 4.0 vs 0.6) and CD62p (8.0+/-3.1% vs 4.1+/-1.8%), CD63 (6.3+/-2.1% vs 3.2+/-1.6%), TXA2 (548+/-85 ng/L vs 390+/-67 ng/L), platelet aggregation rate (43.2+/-10.7% vs 34.8+/-8.1%), thrombosis length in vitro (2.3+/-0.6 cm vs 1.8+/-0.3 cm), CD54 in blood (26.9+/ 6.9% vs 14.4+/-5.1%), CD54 in tissues (51.1+/-6.2% vs 23.1+/-4.1%), Pgp-170 in blood (18.9+/-3.9% vs 10.4+/-2.7%), Pgp-170 in tissues (16.5+/-3.2% vs 10.2+/ 2.3%, P<0.01 or 0.05). CONCLUSION: Based on the characteristics of UC cases in China, different therapy should be given to different types of UC with expected satisfactory results. PMID- 15133866 TI - Rat liver transplantation without preservation of "phrenic ring" using double cuff method. AB - AIM: To develop a double cuff method for rat liver transplantation without preservation of "phrenic ring" to shorten the portal vein clamping time. METHODS: "Phrenic ring" was completely excluded from the donor liver, and end to end anastomosis of suprahepatic inferior vena cava was performed. RESULTS: The portal vein clumping time was shortened to 10.6 min, the successful rate was 83.1%. CONCLUSION: This method can simplify the operation and shorten the portal vein clumping time. PMID- 15133865 TI - Effects of probiotic on intestinal mucosa of patients with ulcerative colitis. AB - AIM: To investigate the effects of probiotic on intestinal mucosae of patients with ulcerative colitis (UC), and to evaluate the role of probiotic in preventing the relapse of UC. METHODS: Thirty patients received treatment with sulphasalazine (SASP) and glucocorticoid and then were randomly administered bifid triple viable capsule (BIFICO) (1.26 g/d), or an identical placebo (starch) for 8 wk. Fecal samples were collected for stool culture 2 wk before and after the randomized treatments. The patients were evaluated clinically, endoscopically and histologically after 2 mo of treatment or in case of relapse of UC. p65 and IkappaB expressions were determined by Western blot analysis. DNA-binding activity of NF-kappaB in colonic nuclear extracts was detected by electrophoretic mobility shift assay (EMSA). mRNA expressions of cytokines were identified by semi-quantitative assay, reverse transcriptase- polymerase chain reaction (RT PCR). RESULTS: Three patients (20%) in the BIFICO group had relapses during 2-mo follow-up period, compared with 14 (93.3%) in placebo group (P<0.01). The concentration of fecal lactobacilli, bifidobacteria was significantly increased in BIFICO-treated group only (P<0.01). The expressions of NF-kappaB p65 and DNA binding activity of NF-kappaB were significantly attenuated in the treatment group than that in control (P<0.05). The mRNA expression of anti-inflammatory cytokines was elevated in comparison with the control group. CONCLUSION: The probiotic could impede the activation of NF-kappaB, decrease the expressions of TNF-alpha and IL-1beta and elevate the expression of IL-10. These results suggest that oral administration of this new probiotic preparation is effective in preventing flare-ups of chronic UC. It may become a prophylactic drug to decrease the relapse of UC. PMID- 15133867 TI - mRNA expression profiling reveals a role of Helicobacter pylori vacuolating toxin in escaping host defense. AB - AIM: To study the immune response of host to Helicobacter pylori VacA. METHODS: The monocyte/macrophage-like U937 cells were infected with Helicobacter pylori vacA-positive strain NCTC 11638 or isogenic vacA-negative mutant. Differentially expressed genes were identified at 2, 6, 10, and 24 h post-infection by cDNA microarray. Differential expressions of some genes were confirmed by Northern blot. RESULTS: More than 100 genes altered their mRNA expression at different time points respectively, many of which were identified to be related to immune evasion. CONCLUSION: VacA is a crucial element for H pylori to escape from host immune defense by means of differentially regulating the expression of some related genes. These genes, previously known or unknown to be involved in the mechanism of immune evasion, deserve further investigation to unearth much more information complicated in the immune response. PMID- 15133870 TI - Journal rejects article after objections from marketing department. PMID- 15133869 TI - Effects of bile reflux and intragastric microflora changes on lesions of remnant gastric mucosa after gastric operation. AB - AIM: To investigate the effects of bile reflux and intragastric microflora changes on lesions of remnant gastric mucosa after gastric operation. METHODS: Concentration of bile acid and total bacterial counts (TBC) in gastric juice were measured in 49 patients with peptic ulcer before and after gastrectomy. One year after the operation, sample of gastric mucosa taken from all the patients were used for histological examination. RESULTS: The concentration of gastric bile acid was significantly increased in group B-I, or B-II and SV+A than that in group HSV (P<0.05-0.01). The abnormal histological changes in the remnant gastric mucosa were more common in the first 2 groups than in the last group. CONCLUSION: The type of gastrectomy can affect bile reflux. The abnormal histological changes in the remnant gastric mucosa are closely related to the elevation of bile acid concentration and increase of TBC in gastric juice. HSV can effectively prevent bile reflux and keep the gastric physiological functions stable. PMID- 15133871 TI - Workplace drug testing. PMID- 15133868 TI - Effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma patients undergoing tumor resection. AB - AIM: To explore the effect of c-myc, Ki-67, MMP-2 and VEGF expression on prognosis of hepatocellular carcinoma (HCC) patients undergoing tumor resection. METHODS: Primary HCC patients underwent tumor resection were retrospectively analysed. The maximum size of the tumor was less than 5 cm, there was only one nodule in each patient. No chemoembolization was performed before resection. They were followed up after resection, and the time of recurrence was recorded. They were divided into 2 groups: group A (15 cases): tumor recurrence within 1 year after tumor resection, and group B (15 cases): with or without tumor recurrence 2 years after tumor resection. Pathological slices were made with tumor wax-sample. Immunohistochemistry staining was performed with c-myc, Ki-67, MMP-2 and VEGF monoclonal antibodies. Staining intensity was quantitatively analysed with a pathological diagram-writing analyzing system. The expressing intensity differences of stained molecules in cancer tissue and para-cancer were analysed. RESULTS: c-myc, Ki-67, MMP-2 and VEGF expressing intensities in cancer tissue in group A were higher than those in group B (P values were 0.010, 0.030, 0.022 and 0.004, respectively), but they were not significantly different in para-cancer tissue in groups A and B (P values were 0.334, 0.343, 0.334 and 0.334, respectively). CONCLUSION: The expression of c-myc, Ki-67, MMP-2 and VEGF in cancer tissue is related to the recurrence of HCC after tumor resection. PMID- 15133872 TI - Does a nurse have a responsibility to report suspicions of drug abuse? PMID- 15133873 TI - What every RN should know about a grievance--Part Two. PMID- 15133876 TI - What are professional boundaries? PMID- 15133878 TI - Meningococcal disease--prevention and intervention. PMID- 15133879 TI - How can I get my unit involved in eliminating health care waste? PMID- 15133880 TI - Gender stratification in management. The World Health Organization 2000. AB - The World Health Organization (WHO) is a global organization that nowadays has integrated gender issues into its policy, programmes and budget. How then is the state of affairs in the area of gender equity at the ultimate governing bodies of the modern WHO? This study aims to assess the representation of women and men and their promotion within the supreme decision-making bodies of the WHO during the year 2000. Information sources used are the official and confirmed protocols of the 53rd World Health Assembly (WHA) in 2000 and of the two Executive Board (EB) meetings of the corresponding year. A descriptive quantitative content analysis approach is used exclusively. The present study demonstrates strikingly skewed gender distribution, with men substantially at an advantage numerically in the prominent positions at the WHA 2000. Additionally, men also hold an advantage in terms of being promoted to leading positions within the bodies examined, notably all upgraded chairs of the EB during 2000. However, the formerly male-dominated supervisory positions of the WHO are, these days, challenged by women having been elected at the very top of the WHO. The present study stresses the need to elaborate a qualitative research design to advance the understanding of the social construction of gender in supreme governing positions of the modern WHO. PMID- 15133881 TI - Creating diversity in the healthcare workforce. The role of pre-registration nurse education in the UK. AB - It has been suggested that the introduction of first the diploma pre-registration course and then the three-year degree pre-registration course are likely to have an impact upon the diversity of the future nursing workforce in the UK. Using findings from three large-scale projects, this paper assesses the nature of these changes by comparing the diversity within recently qualified cohorts of graduate, diplomate and certificate-qualified nurses. Findings indicate little difference between the three groups in terms of gender and ethnic origin. A higher proportion of graduates had academic qualifications, which were generally higher than the qualifications held by the other groups of nurses. However, diplomates were the most diverse group overall, being older and more likely to have children, and also having greater previous working experiences. The overall conclusion is that, despite qualification differences, the level of diversity amongst degree-qualifiers was a return to the lower levels associated with qualifiers from certificate courses. Thus the introduction of degree courses may dilute the relative success of the diploma course in terms of the recruitment of a more diverse workforce. The findings are considered in the context of current debates about the future shape of nurse education in the UK. PMID- 15133882 TI - The impact of management programs on physicians' work environment and health. A prospective, controlled study comparing different interventions. AB - Female physicians have less influence over their daily work conditions and exhibit slower career advancement as compared to their male colleagues. The aim of this study is to assess the impact on individual and organizational well being from different kinds of management programs. Female physicians participating in management intervention programs were compared with a reference group of matched physicians and sickness absenteeism was significantly lower in the intervention group. No significant differences were found between the groups with regard to career advancement, individual, organizational and professional well being. Health care organizations spend a substantial amount of resources on management programs in order to improve leadership, autonomy and the work-environment of physicians in times of increasing discontent among this key group of health care employees. Our study indicates some beneficial health effects from structured management programs but there is a need to develop and assess the efficacy of these programs further. PMID- 15133883 TI - Perceived potential risk factors in child care. AB - This study is based on semi-structured interviews focusing on staff members' opinions about potential risk factors that could threaten patient safety. The aim was to acquire more in-depth knowledge about the causes of patient injuries. The study, which was conducted at a children's hospital, has a qualitative approach that is influenced by the critical incident technique. A total of 28 persons were interviewed. Analysis of the data resulted in five qualitatively differentiated categories of potential risk factors: a large influx of patients, a lack of professional experience, a lack of inter-professional communication and cooperation, and deficiencies related to work hours and to the physical environment. The results reflect a complex picture where the risks, as described by the informants, can either alone or in concert directly or indirectly affect the individual in the practice of his or her profession or contribute to a mistake. PMID- 15133884 TI - Innovation and funding specialist services. Cochlear implantation. AB - This paper traces the innovative development of the Nottingham Cochlear Implant Programmes. The paediatric programme was the first to be established in the UK in 1989 and remains the largest programme in the UK today, whilst the adult programme developed later, in 1994. The first section of the paper describes trends in service development whilst the second section makes detailed reference to the history of funding arrangements which enabled the programme to become established. The third part of the paper examines the (de)merits of locality purchasing versus centralised purchasing for specialist services, using cochlear implantation as way of illustration. The paper aims to provide an informative history of the development of the service in Nottingham and from this background create debate as to the most appropriate future funding mechanism for cochlear implantation in particular and specialist services in general. PMID- 15133886 TI - Effects of omega-3 fatty acids on arrhythmogenic mechanisms in animal and isolated organ/cell culture studies. PMID- 15133885 TI - Health effects of omega-3 fatty acids on asthma. PMID- 15133887 TI - Effects of omega-3 fatty acids on cardiovascular risk factors and intermediate markers of cardiovascular disease. PMID- 15133888 TI - Effects of omega-3 fatty acids on cardiovascular disease. PMID- 15133889 TI - Regionalization of bioterrorism preparedness and response. PMID- 15133890 TI - Effects of omega-3 fatty acids on lipids and glycemic control in type II diabetes and the metabolic syndrome and on inflammatory bowel disease, rheumatoid arthritis, renal disease, systemic lupus erythematosus, and osteoporosis. PMID- 15133891 TI - [Studies on expression and location of VEGF protein in rat testis and epididymis]. AB - In order to investigate the function of vascular endothelial growth factor (VEGF) in the male reproductive system, we examined the expression and localization of VEGF protein in adult rat testis and epididymis with the immunohistochemical method and PAS stain and western blots. The results showed that the VEGF protein (about 45 kD) was present in the rat testis and epididymis. The VEGF immunoreactive particles were localized in the cytoplasm of spermatids, Sertoli cells and Leydig cells with negatively stained nuclei. The expression pattern of the spermatids was different in the 14 stages of epithelial cycle, and the strong immunopositive reaction was mainly in the forming/formed acrosome of round/elongated spermatids and spermiogenic residual bodies. In the epididymis, the region-specific and cell-specific expression and localization of VEGF protein was shown in the epididymal epithelium. In the initial segment, the VEGF protein was presented in the whole epithelial cells and the immunoreactive particles were localized in the supranuclear region of the cells. Except for a few principal cells and basal cells, counter-staining of PAS stain proved that the immunoreactive cells were clear cells in the caput, corpus, and cauda epididymis. And the immunoreactive particles were localized in the different region of the cytoplasm of clear cells, some were in the supranuclear region, some were filled in the whole cytoplasm, and some were secreted out of the cells along the free border. The positive acrosomes of the spermatozoa were only seen in the lumen of the proximal epididymis. No expression was detected in the interstitial blood vessels and the capillaries of the testis and epididymis. The present results indicate that VEGF protein may play an important role in the process of the spermatogenesis and spermiogenesis, especially in the acrosomal developing process. VEGF may also be related to the sperm maturation in the epididymis. PMID- 15133892 TI - [The tempo of meiosis of goat oocytes]. AB - The meiotic progression of goat oocytes from follicles of different diameters was investigated in this study. The results were summarized as follows: (1) The in vitro meiotic maturation capacity was different among oocytes from follicles of different diameters. And thus oocytes from < or = 0.5 mm follicles were unable to resume meiosis; oocytes from 0.8-1.2 mm follicles were capable to resume meiosis, but could develop only to MI stage (60% at 24 h); oocytes from 1.5-5 mm follicles had acquired full-meiotic maturation capacity and 91% of them developed to M II stage at 24 h of culture. (2) The percentage of oocytes with intact-germinal vesicles from 1.5-5 mm follicles decreased significantly during 2-8 h of in vitro maturation and the decrease was even more rapid during 4-6 h of culture (from 60% to 19%, p < 0.0005). The percentage of oocytes at M I-stage increased from 24% to 61% during 6-12 h of in vitro maturation, and it then decreased. By 24 h of culture, only 2% oocytes remained at M I-stage. Twenty one percent of the oocytes in this group developed to M II-stage at 16 h of culture, and by 24 h of culture, 91% were at M II-stage. (3) Statistic analysis of the meiotic progression (the duration of each cell cycle stage) of oocytes from 1.5-5 mm follicles showed that GV stage lasted from 0 to 3 h of culture, prometaphase-I stage was from 3.0 to 7.0 h, metaphase-I stage was from 7.0 to 14.6 h, anaphase-I/telophase-I was from 14.6 to 18.4 h and metaphase-II stage lasted from 18.4 to 24 h. (4) Whether the oocytes capable of GVBD and entrance of M I developed to M II, the timing of meiotic progression prior to M I was similar. In summary, our results provided necessary data for studies on the mechanisms and control of meiosis in mammalian oocytes. PMID- 15133894 TI - [Immunohistochemical analysis and immuno-gold localization of ECBP21 in Angelica dahurica]. AB - ECBP21 is a calmodulin binding protein (CaMBP) purified from extracellular extracts of suspension-cultured cells of Angelica dahurica, and it is the first reported extracellular CaMBP in plant kingdom. We have recently cloned the full length cDNA for ECBP21. In this work, using recombinant ECBP21 we prepared rabbit antiserum with high specificity and high titer against ECBP21, and investigated the organ-specific distribution of ECBP21 in Angelica dahurica. ECBP21 was found in all organs examined, particularly abundant in the leaves flowers, and raches, and less in the roots. It was also found in all cells examined, and particularly enriched in the cell wall. These data support the notion that ECBP21 is specifically localized extracellularly, and imply that it may be involved in plant growth and development. In addition, using immunogold transmission electron microscopy method, we studied the subcellular localization of ECBP21 in rachis cells of Angelica dahurica. The results indicated that the ECBP21 was mainly localized in cell wall; this provided a direct evidence of the extracellular existence of ECBP21. PMID- 15133893 TI - [Changes of thioredoxin mRNA level in neurons with mitochondrial dysfunction insulted by H2O2]. AB - Oxidative stress and mitochondrial deficiency have been considered to be associated with the mechanisms of many neurodegenerative diseases. Sodium azide (NaN3) is a special inhibitor of mitochondrion cytochrome c oxidase (COX), which can be used to mimic neuronal damage induced by mitochondrial deficiency. In this experiment, the neurotoxic effects of H2O2 on primary cultured neurons and NaN3 induced mitochondrial dysfunctional neurons were detected by means of cell viability measurement (MTT) and analyzed through morphological observation. Furthermore, the changes of thioredoxin mRNA level in both normal and abnormal cultured neurons insulted by H2O2 were analyzed by semiquantitative RT-PCR in order to explore the role of Trx, an important redox regulatory protein, in modulating the process of neuronal injury. It was found that mitochondrial dysfunctional neurons could be damaged by H2O2 in a dose- and time-dependent manner and the expression of Trx decreased during certain dose (0-200 mumol/L) and time (0-4 h) of H2O2 treatment. But in normal neurons the effects of H2O2 treatment seemed to be less evident. It suggests that in neurons with mitochondrial dysfunction, the redox modulation of thioredox seems to be more prominent. PMID- 15133895 TI - [Effect of methyl viologen on the Ca(2+)-ATPase activities of membrane vesicles in roots of rice seedlings]. AB - The cold tolerance of rice (Oryza sativa L.) seedlings could be markedly improved by mild oxidative pretreatment (10 mumol/L methyl viologen (MV). The enhancement of chilling resistance induced by MV pretreatment could be inhibited by Ca(2+) chelate ethyleneglycol-bis-(beta-aminoethylether) N,N-tetraacetic acid (EGTA, 10 mmol/L) and the calmodulin inhibitor chlorpromazine (CPZ, 0.5 mmol/L). The Ca(2+) ATPase activity of root plasmolemma and tonoplast membrane as well as the Fe(CN)6(3-) reduction of plasmolemma were also enhanced by MV pretreatment. However, the above MV pretreatment effect could be inhibited by EGTA and CPZ, respectively. In vitro, Ca(2+)-ATPase activities in the two types of membrane vesicles were reduced by reactive oxygen species (H2O2, O2-., .OH). Results indicated that the enhancement of chilling resistance in rice seedlings with MV pretreatment was likely related to the effective activation of Ca(2+)-ATPase induced by calcium messenger of CaM, and the improved cold tolerance of the activities might be associated with the augmentation of cellular membrane protective ability caused by the MV pretreatment. PMID- 15133897 TI - [Comparative studies on proteins of cytoplasmic male-sterile wheat and its maintainer by 2D-PAGE in Triticum aestivum]. AB - By using 2D-PAGE techniques, protein compositions have been studied at different developmental stage leaf (seedling, tillering, shooting and booting stages) and pollen mother cells (PMC) (meiotic, 1-nucleus and 2-3 nuclei stages) in male sterile wheat and its maintainer. The results indicated that a specific protein (33KD/pI6.3) identified at the shooting and booting stage leaves in male-sterile wheat, but this protein species has not been identified in the male fertile wheat. Four protein species, 53KD/PI5.5, 50KD/PI5.7, 48KD/PI5.6, 20KD/PI7.5 were detected at the 2-3 nuclei stages, which was thought key stage of pollen fertility lost in male-sterile wheat, and these protein species were not be seen in male-fertile wheat. According above experiment results, five protein species which detected in leaf and PMC might participate in fertility regulation and related to the male-sterility of male-sterile wheat. PMID- 15133896 TI - [The starchy endosperm denucleation by a process of programmed cell death during rice grain development]. AB - The cellular ultrastructural morphology, grain filling ratio and activity of related enzymes during denucleation development stage in starchy endosperm cell of an Indica rice variety Zhongxian 8836 were observed and analyzed. The endosperm cellularization was completed at about the 3rd day after flowering (DAF). At 5 DAF a few endosperm cells initiated denucleation development. At 13 DAF almost all the starchy endosperm cells completed their denucleation. The nuclear degradation is the first stage of programmed cell death (PCD) in starchy endosperm. It occurred unsynchronously among different starchy endosperm cells. The nuclear degradation of starchy endosperm cell during denucleation development stage showed not only morphological features commonly observed in animal and plant PCD, but also some unique characteristics. Mitochondria degeneration was observed along with nuclear degradation, indicating there were interrelations between the two processes. Enzymes related to PCD, such as super-oxide dismutases (SOD) and catalase (CAT), as well as enzymes related to starch synthesis, such as ADP-glucose pyriphosphorylase (AGP), soluble starch synthase (SSS) and starch branching enzyme or Q-enzyme, showed very high activity during the denucleation development stage. Maximum grain filling rate and grain weight increase were also achieved in the denucleation developing stage of most starch endosperm cells. The denucleated cell remained alive in the developing endosperm, keeping its normal metabolisms, the synthesis and accumulation of starch and storage proteins. However, enzyme activities and grain filling rate were apparently dropped to a lower level after denucleation. The starchy endosperm cell finally completed its PCD process when it was completely filled with reserves. Evan's blue staining indicated that cell death occurred unsynchronously among the starchy endosperm cells with initiation points randomly distributed in the endosperm tissue. PMID- 15133898 TI - [Changes of some immune-mediators in CCl4-induced liver injury mice]. AB - Changes of some immune-mediators in liver tissue homogenate and plasma of liver injury mice injected with CCl4 and its mechanism in the liver injury process induced by CCl4 were studied. Thirty healthy mice with half of female and male were chosen and divided randomly into two groups which were control group (group C) and CCl4-injecting group (group CCl4). The liver injury was induced by abdominal injection of CCl4 (0.07 ml/100 g body weight) on every other day over six weeks. The blood and liver were collected at the 2nd, 4th and 6th week, respectively, and liver homogenate cAMP, cGMP levels and MDA concentration as well as plasma IL-2 and TNF-alpha levels were determined. The results show that during the entire experimental period, cAMP level decreased or markedly decreased in group CCl4. After the 2nd week of CCl4-injecting, cGMP was lower or significantly lower in group CCl4 than in group C; cAMP/cGMP ratio tended to drop, and was low or significantly low to group C; and MDA content was significantly higher in group CCl4 than in group C. During the whole experimental period, a marked decrease of plasma IL-2 in group CCl4 was seen, and plasma TNF alpha of group CCl4 was superior to that of group C. It is suggested that the violent changes of immune-mediators including cAMP, cGMP, TNF-alpha and IL-2 etc induced by CCl4 may play an important role in the induction of liver injury in mice. PMID- 15133899 TI - [A study on organotypic spinal cord slice culture]. AB - This study was aimed at developing a method of organotypic culture of spinal cord slice. The slice culture was prepared using lumbar spinal cord from 8-day-old rat. The survival of alpha-motor neuron was evaluated by the morphological observation and by monoclonal antibody SMI-32, a nonphosphorylated neurofilament marker and immunohistochemical staining. Lactate dehydrogenase (LDH) level in culture medium was also measured. The result showed that the spinal cord explant could be maintained in the culture for more than 2 months with excellent cellular organization and stable population of ventral motor neurons. The level of LDH at different culture times had no significant difference. Organotypic spinal cord slice culture may provide an effective method for studying physiological and pathological changes, and neuroprotection of spinal cord. PMID- 15133900 TI - [The study of sperm cryopreservation in kunmin (KM) mouse]. AB - Spermatozoa of 10, 15 and 20-wk-old KM mice were frozen and stored at--196 degrees C. After thawing, intact superovulation oocytes were inseminated in vitro with relative high motility frozen/thawed mouse spermatozoa. Some 2-cell embryos obtained by in vitro fertilization were transferred to pseudopregnant recipients and 47%-56% of them developed into live-born offspring. Some 2-cell embryos were incubated in vitro and 57% of them progressed to the blastocyst stage. Others were cryopreserved by simple vitrification procedure, and most of them (about 80%) were morphologically normal after warming, and 50% of the cryopreserved embryos transferred to recipients had developed into live pups. These results indicated that there was no significant difference when compared with embryos obtained from oocytes inseminated with fresh sperm. PMID- 15133901 TI - [Carpel development in male flowers of cucumber (Cucumis sativus. L)]. AB - Carpel development in cucumber male flowers was studied by morphological, histochemical and isoenzyme electrophoretic analyses. The results showed that: (1) cell number of the carpel in male flowers increased continuously during the development of male flowers, and the carpel in male flowers was abundant in RNA content; (2) the carpel in male flowers at latter developmental stage was differentiated, and a placenta-like structure was formed in the carpel of male flowers during anthesis, while the ovule did not appear as that of mature female flowers; (3) The POD and esterase isoenzyme electrophoretic profiles of carpel varied from the development of male flowers, which further indicated that the carpel in mature male flowers was no longer at the stage of primordium. It could be deduced from the results that carpel of cucumber male flowers develops continuously in a distinct pattern with that of female flowers during sex expression process. PMID- 15133902 TI - [Selection of scaffolds of rat hepatocytes in three-dimension culture under simulated microgravity]. AB - In order to investigate the biocompatibility, degradation and inflammatory reactions of poly benzyl glutamate (PBLG), poly lactic-co-glycolic acid (PLGA) and fibrin gel used as the scaffolds co-cultured with the hepatocytes of rats under simulated microgravity conditions in rotating wall vessel bioreactor (RWVB). PLGA is low biocompatibility, PBLG is medium biocompatibility with rat hepatocytes. Fibrin gel is high biocompatibility, no toxic degradation and no inflammatory reactions. Fibrin gel can be used as three dimensional scaffolds of rat hepatocytes under simulated microgravity. PMID- 15133903 TI - [The origination and action of the hepatic stems cells]. AB - Oval cells are small cells with scant cytoplasm and ovoid-shaped nuclei. These cells, probably deriving from the bone marrow or the cell population associated with the bone marrow, are activated hepatic stem cell. The morphological characteristic of the hepatic stem cells is similar to the oval cells and its biochemistry marker is c-kit/CD45/TRE19. In this paper, the knowledge about the hepatic stem cells and their functions responsible for liver development, liver regeneration, carcinogenesis and relation to other cell are reviewed. PMID- 15133904 TI - [Cardiac excitation-contraction coupling mediated by Ca2+]. AB - Cardiac excitation-contraction coupling is the process which links electrical excitation of the cardiac myocytes with heart contraction. Of the ions involved in cardiac excitation-contraction coupling, calcium is considered to be the essential and crucial factor. In this short communication, we will briefly illustrate how calcium is involved in excitation-contraction coupling in order to better understand the heart function. PMID- 15133905 TI - New health worker credentialed in Ohio. PMID- 15133908 TI - [Effects of extrahypothalamic brain lesions on testicular function in rats--with special emphasis on asymmetry]. AB - INTRODUCTION: The aim of our studies was to investigate the involvement of extrahypothalamic brain structures in the control of testicular functions with special emphasis on the effect of right- and left-sided structures. MATERIAL AND METHOD: We performed lesion of the insular cortex, the amygdala, interrupted part of nerve fibers to and from the insular cortex, and cut the major commissural pathway of the brain the corpus callosum in adult male rats and studied the effect of the interventions on testicular steroidogenesis, serum testosterone and gonadotrop hormone concentrations. RESULTS: Following lesion of the insular cortex on the right side serum testosterone level and steroidogenesis of the testes decreased (in the case of the left testis the difference was significant). Similar lesion on the left side did not change the parameters studied. Both right and left-sided lesion induced a significant increase in serum LH concentration. The effect was more pronounced after right-sided lesion. Interruption of nerve fibers above the amygdala by a paramedian sagittal knife cut on the right or on the left side resulted in opposite effect on testicular steroidogenesis: right sided intervention increased while left-sided one reduced testosterone secretion. Only left-sided cut influenced (decreased) serum testosterone level. There was no changes in LH concentration. Both right- and left-sided lesion of the amygdala induced a significant decrease in basal testosterone secretion in vitro of both testes and in serum testosterone level. However, serum LH concentration decreased only after left-sided surgery. Interruption of the corpus callosum in animals with left-sided orchidectomy induced a significant rise in steroidogenesis of the remaining (right) testis. Both sham surgery and callosotomy combined with left orchidectomy resulted in a significant increase in serum FSH level. CONCLUSION: Results of our studies suggest that extrahypothalamic brain structures and interventions influence endocrine functions of the testis through the hypothalamo hypophyseal-testicular axis and by a direct neural route. Certain components of the regulatory system exhibit functional asymmetry. PMID- 15133909 TI - [Neuroprotection in brain ischemia--doubts and hopes]. AB - In ischaemic stroke the two major potential therapeutic strategies are aimed at either improving cerebral blood flow or directly interacting with the cytotoxic cascade--a large body of evidence gained from animal studies is in support of them. In clinical trials direct neuroprotection by blocking the neurotoxic cascade remained ineffective, although there are several clinical trials still in progress. We summarize the experimental data and present the results of clinical trials and also discuss why so many drugs, which were effective in animal studies, failed in human trials. It is emphasized, that 1. in most animal studies the reduction of infarct size, i.e. the amount of saved penumbral tissue, was the outcome measure, whereas neurological function remained unassessed; 2. the recovery of intellectual performance and higher cortical functions are of major importance in the future quality of life in stroke victims; however, it is impossible to examine these parameters appropriately in animal studies; 3. in many clinical trials the patient population was rather heterogenous and low in number, the study protocol was not optimal and the critical analysis of the subacute and chronic phase was lacking or insufficient. We present the major experimental stroke models, discuss their similarities, differences and limitations as compared to the human pathophysiological processes. The pitfalls of extrapolating data from animal studies to clinical practice are also summarized. The complex network of functional and morphological intercellular connections, the long timescale of neurotoxic and reparative events and the lessons learned from clinical trials suggest, that the use of drug combinations (therapeutic cocktails) targeting multiple steps of the neurotoxic cascade would hopefully result in more effective treatment of ischaemic stroke. Strategies to facilitate brain plasticity and regeneration is an additional promising tool to enhance recovery in brain ischaemia. PMID- 15133910 TI - Serum IgG and IgM ganglioside GM1 antibodies in patients with multiple sclerosis. AB - INTRODUCTION: In order to obtain more information concerning the pathogenic significance of ganglioside GM1 in multiple sclerosis serum polyclonal IgG and IgM antibodies to GM1 were evaluated in multiple sclerosis patients with relapsing-remitting and secondary progressive forms of the disease. PATIENTS AND METHODS: The evaluated sera were from 55 patients with clinically definite multiple sclerosis and from 20 healthy subjects. Forty-two of patients were with relapsing-remitting and 13 with secondary progressive multiple sclerosis. Antibodies to GM1 were measured using a modification of the enzyme-linked immunosorbent assay technique of Mizutamari et al (1994). RESULTS: A statistically significant difference of serum IgG antibody titres to GM1 was found between the healthy subjects and the multiple sclerosis patients with relapsing-remitting form of the disease (p = 0.04), as well as of serum IgG antibody titres to GM1 between the patients with relapsing-remitting multiple sclerosis in relapse and in remission (p = 0.01). CONCLUSION: Bearing in mind the heterogeneity of multiple sclerosis, the pathogenic significance of serum antibodies to GM1 should be interpreted concerning the precise clinical form of the disease and not the whole group of MS patients. The findings in this study argue for the possible involvement of ganglioside GM1 in the pathogenesis of demyelination in relapsing-remitting multiple sclerosis. PMID- 15133911 TI - Epilepsy caused by retrosplenial tumor. AB - We present a patient in whom retrosplenial tumour was associated with epileptic symptoms characterized by complex partial seizures and widespread interictal and ictal epileptiform EEG abnormalities. The patient had verbal memory deficit symptoms as well. After surgical removal of the tumour (oligoastrocytome) the clinical symptoms and EEG signs disappeared. The characteristics of our patient demonstrate the possible role of the retrosplenial area in widespread epileptic symptoms and in the regulation of secondary bilateral synchrony, in addition to its recently described importance in the memory functions. PMID- 15133912 TI - [New theories of the self]. PMID- 15133913 TI - [Report from the Meeting of the Committee of European Neurology Specialty Education]. PMID- 15133914 TI - Neurological aspects of some sleep disorders. AB - My aim is to examine the relation between some sleep disorders and neurological diseases; to analyse their mutual interactions in order to achieve new practical data for clinical use. In the theoretical part I summarise some main points of sleep physiology concentrating on the associations of sleep regulation and neurological diseases. In my examinations, besides clinical methods, the most important tools used are sleep analyses performed by polysomnography and MESAM IV as well as brain imaging methods. To assess clinical state of my stroke patients I utilised NIH Stroke Scale. I found pathological sleep apnoea frequency in more than half of the patients in any type (bleeding/infarction) of acute stroke. In a prospective study, sleep apnoea parameters remain permanent during 3 months in the ischaemic group; on the other hand, sleep apnoea improves during follow up after brain haemorrhages. I showed pathological sleep apnoea frequency in myasthenia gravis among male patients without daytime respiration complaint. I looked for the link between the mechanism of the sleep disorder and the underlying organic lesion in two cases. In this analyses I took into account the function of the affected structure in sleep regulation. I found a basal forebrain tumour, affecting sleep regulating centres underlying severe insomnia and I suggest a neuro-vascular compression of the lateral preoptic area of the hypothalamus being the reason of sleep related painful erection, a parasomnia of unknown origin. PMID- 15133915 TI - Lessons from SARS: challenges for the international nursing research community. PMID- 15133916 TI - Health promotion and literacy: implications for nursing. PMID- 15133917 TI - Adolescent constructions of nicotine addiction. AB - The purpose of this qualitative study was to extend our understanding of how adolescents view nicotine addiction. This secondary analysis included 80 open ended interviews with adolescents with a variety of smoking histories. The transcribed interviews were systematically analyzed to identify salient explanations of nicotine addiction. These explanations presuppose causal pathways of nicotine exposure leading to addiction and include repeated use, the brain and body "getting used to" nicotine, personal weakness, and family influences. A further explanation is that some youths pretend to be addicted to project a "cool" image. These explanations illustrate that some youths see themselves as passive players in the formation of nicotine addiction. The findings can be used in the development of programs to raise youth awareness about nicotine addiction. PMID- 15133918 TI - Multiple intervention research programs in community health. AB - The authors describe an organizing framework for multiple interventions in community health. The framework provides a foundation for programmatic research on multiple interventions and poses critical questions that need to be addressed in the next generation of research in this field. Multiple intervention programs are characterized by the use of multiple strategies targeted at multiple levels of the socio-ecological system and delivered to multiple target audiences. Consequently, they complement the growing literature on the broad determinants of health and health promotion. The authors describe a 4-stage framework and identify gaps and challenges in this field of research. There are 5 key research areas requiring concerted action; researchers must: examine nested determinants, develop integrated conceptual frameworks, examine ways to optimize synergies among interventions, describe spin-offs from multiple intervention programs, and monitor the sustainability of their impact. PMID- 15133919 TI - Multilevel health promotion research: conceptual and analytical considerations. AB - Health promotion research is often conceptualized through the use of socioecological frameworks. This results in data or variables associated with multiple levels such as individual, community, and provincial. These data are nested, or clustered. In other words, multilevel health promotion research is based on the idea that community influences health, above and beyond one's individual characteristics or behaviours. These contextual effects can be analyzed rigorously using multilevel modelling (MLM), thus determining whether contextual effects are truly derived from context or are the result of residents' social profile. MLM also facilitates examination of cross-level interaction effects. The authors discuss conceptual and methodological issues related to multilevel research. While multilevel pathways to health outcomes have been suggested at the conceptual level, analytical techniques that produce only average overall effects fail to reveal the various other influences on health behaviour. PMID- 15133920 TI - A brief intervention to increase physical activity in sedentary working women. AB - The purpose of this study was to determine whether a brief, tailored counselling intervention is effective for increasing physical activity in sedentary women when delivered in the workplace. This prospective randomized trial used a brief intervention based on the Transtheoretical Model. The intervention group received health screening, a brief intervention, and, 2 weeks later, a booster telephone call from a nurse practitioner. The intervention was tailored to each woman's reported exercise behaviour. The 134 women randomized to the intervention were compared with 153 women in a control group who received health counselling not tailored to their exercise behaviour and no telephone call. Six weeks later, the intervention group, when compared to baseline, had significantly improved their physical activity, increasing their amount of weekend physical activity as well as minutes walked for exercise, on errands, total walking, and total daily blocks walked. When compared to controls, they showed significantly greater gains. This test of a brief, tailored strategy provides a critical contribution to the search for efficient, effective ways for nurses to deliver workplace health promotion interventions. PMID- 15133921 TI - Nurses as caregivers of elderly relatives: negotiating personal and professional boundaries. AB - Recent changes in patterns of care provision for the elderly have led to an increasing reliance on family care. Although caring has been found to be a central and common feature of the personal and professional lives of many women, this paper discusses the challenges faced by women who provide care in both their work and their family lives ("double-duty caregivers"). The author argues that the separation of paid caregiving and unpaid family caregiving in the conceptualization of elder care is problematic, particularly for health-care professionals. Findings from a qualitative study with registered nurses providing care to elderly relatives revealed that these women are located at the juncture of public and private domains of caregiving, where they must constantly negotiate the boundaries between their professional and personal caregiving roles. The findings highlight the need to explore the interface between women's family and work lives and the need for policies that promote the health of double-duty caregivers. PMID- 15133922 TI - Conceiving action, tracking practice, and locating expertise for health promotion research. PMID- 15133923 TI - Mediating complaints against nurses: a consumer-oriented educational approach. AB - A participatory evaluative method was used to assess the effectiveness of mediation as carried out by the College of Nurses of Ontario. Qualitative methods were used to examine 34 cases between 1994 and 1998, of which 23 had been successful and 11 aborted. For purposes of comparison, the researchers developed a template of interviews with College personnel and documents, incorporating the College's philosophy and expectations of the process. Semistructured interviews were conducted with 44 participants in the mediation process. In addition, focus group sessions were held with Investigators and Practice Consultants. The data were analyzed using the template and themes were generated. The process was found to be stressful for all parties but was also found to be educational, to address system complaints, and to achieve initial goals. The College was found to be powerless to demand system reforms and to be dependent on the cooperation of each facility. PMID- 15133924 TI - Choosing a model of care for patients in alternate level care: caregiver perspectives with respect to staff injury. AB - The population of alternate level care (ALC) patients utilizing acute-care hospital resources inappropriate to their needs is growing. The purpose of this study was to explore how the care of ALC patients was managed at 4 acute-care facilities in the Canadian province of British Columbia and to examine how this care impacts on outcomes of staff injury. Interviews were conducted to identify and characterize the different models of ALC. Injury outcomes for all caregivers were obtained (n = 2,854) and logistic regression conducted to compare staff injuries across ALC models. Injured workers were surveyed regarding their perceptions of injury risk and ALC. Five ALC models were identified: low-mix, high-mix, dedicated ALC units, extended care units, and geriatric assessment units. The risk for caregiver injuries was lowest on dedicated ALC units. These findings suggest that acute-care facilities faced with a growing ALC population should consider creating dedicated ALC units. PMID- 15133925 TI - PORT--Psychosocial Oncology Research Training: a newly funded strategic initiative in health research. PMID- 15133926 TI - Synthesis and characterization of albumin binding surfaces for implantable surfaces. AB - Implanted medical devices are of increasing importance in the practice of medicine and it is estimated that more than 3-million people in the United States have long-term implants like vascular grafts, pacemakers, catheters and joint replacements. Even though most biomedical polymers employed are relatively inert, unreactive, and non-toxic, most implant materials when contacted with blood invoke the activation of blood cells as well as the plasma proteolytic enzyme systems. In order to overcome aforementioned problems, attempts have been made to modify the surface chemistry of the blood-contacting biomaterials so that they become thromboresistant. One potential strategy to address the problem of the chaotic adsorption of proteins to medical devices and surfaces is to create a layer of albumin as a "barrier protein cover", as a thin layer of albumin has been reported to minimize adhesion and aggregation of platelets. By grafting peptides and synthetic ligands, that specifically bind human serum albumin, on an epoxide functionalized surface, we have generated surfaces that exhibit enhanced albumin binding over other serum proteins. We have characterized the surface using infra-red ellipsiometry and quantified protein adsorption via specific ELISA assays and gel electrophoresis. PMID- 15133927 TI - Morphology and growth of murine cell lines on model biomaterials. AB - All biomaterial implants are assaulted by the host "foreign body" immune response. Understanding the complex, dynamic relationship between cells, biomaterials and milieu is an important first step towards controlling this reaction. Material surface chemistry dictates protein adsorption, and thus subsequent cell interactions. The cell-implant is a microenvironment involving 1) proteins that coat the surface and 2) cells that interact with these proteins. Macrophages and fibroblasts are two cell types that interact with proteins on biomaterials surfaces and play different related, but equally important, roles in biomaterials rejection and implant failure. Growth characteristics of four murine cell lines on model biomaterials surfaces were examined. Murine monocyte macrophages (RAW 264.7 and J774A.1), murine macrophage (IC-21) and murine fibroblast (NIH 3T3) cell lines were tested to determine whether differences exist in adhesion, proliferation, differentiation, spreading, and fusion (macrophage lineages only) on these surfaces. Differences were observed in the ability of cells to adhere to and subsequently proliferate on polymer surfaces. (Monocyte-) macrophages grew well on all surfaces tested and growth rates were measured on three representative polymer biomaterials surfaces: tissue culture polystyrene (TCPS), polystyrene, and Teflon-AF. J774A.1 cultures grown on TCPS and treated with exogenous cytokines IL-4 and GM-CSF were observed to contain multinucleate cells with unusual morphologies. Thus, (monocyte-) macrophage cell lines were found to effectively attach to and interrogate each surface presented, with evidence of extensive spreading on Teflon-AF surfaces, particularly in the IC-21 cultures. The J774A.1 line was able to proliferate and/or differentiate to more specialized cell types (multinucleate/dendritic-like cells) in the presence of soluble chemokine cues. PMID- 15133929 TI - Analysis of regenerated amine-reactive polymer microarray slides. AB - Reactive polymer-coated microarray substrates based on N-hydroxyl succinimide (NHS) chemistry lose their bio-immobilization reactivity to probe nucleophiles over time, both in use and in storage, due to their intrinsic hydrolytic instability. Poor DNA and protein probe immobilization efficiency is often observed with routine microarray printing conditions, with accompanying reliability and stability issues for assay. We report a one-step reaction to regenerate NHS-reactive chemistry in situ on these microarray polymer surfaces with simple, straightforward reaction chemistry. Surfaces regenerated with this method perform equal to or better than freshly prepared slides in print immobilization of oligonucleotide probes functionalized with primary amine reactive groups. DNA probe specific and non-specific surface binding as well as target hybridization were analyzed using both fresh and regenerated slides. Commercial microarray substrates appear to retain DNA probes with both substantial covalent immobilization and some non-specific adsorption to the surface. PMID- 15133928 TI - Surface modification of UHMWPE for use in total joint replacements. AB - To create a hydrophilic, lubricious, more wear-resistant UHMWPE bearing, a novel hyaluronan (HA) derivative and novel UHMWPE-hyaluronan composite were developed. HA was silylated to increase its hydrophobicity and compatibility with UHMWPE. The sily1 HA rapidly diffused into the connected pores of UHMWPE preforms in xylenes solution, and fixed within UHMWPE and on its surface after crosslinking. A micro-composite was obtained after hot-pressing the porous preform. The presence of HA film on the composite surface has been demonstrated through X-Ray photoelectron spectroscopy (XPS) analysis and Toluidine Blue O (TBO) dye assay. The aqueous contact angles of micro-composite samples were significantly lower compared with UHMWPE control samples, and the samples processed with hydrolysis prior to final molding were superior to those processed with hydrolysis after molding. PMID- 15133930 TI - Influence of muscle contraction on whiplash kinematics. AB - It is unclear whether reflexive muscle contraction in unaware occupants can alter spinal kinematics to mitigate injury in the unaware occupant subjected to whiplash loading. Whiplash injury likely occurs during the non-physiologic S curvature phase of spinal kinematics, present during the first 100 msec after the initiation of T1 acceleration. Experimental investigations using human volunteers have reported 45 to 60 msec delays prior to electrical activity of the sternocleidomastoid. The effects of reflexive contraction of the neck muscles were investigated using a validated head-neck computational model consisting of head, cervical spine, and first thoracic vertebra. Intervertebral discs. spinal ligaments, and facet joints were modeled using discrete elements. Passive and active musculature were incorporated using the Hill-type muscle model. The computational model was subjected to 2.6 m/sec rear impact velocity, applied to T1. Reflexive muscle contraction in the unaware occupant model was incorporated using a 54-msec muscle delay, 13-msec electromechanical delay, and an 81-msec muscle rise time. Results of the unaware occupant model were compared to the model exercised without muscle contraction. Reflexive muscle contraction altered segmental angulations by less than 10% and facet joint capsular ligament distractions by less than 16% during the time of maximum S-curvature. At the C5 C6 and C6-C7 levels, muscle contraction increased capsular ligament distractions. Due to the nominal affect of reflexive muscle contraction on segmental angulations and facet joint capsular ligament distractions during S-curvature, it is unlikely that this contraction can alter the cervical kinematics responsible for whiplash injury. PMID- 15133931 TI - NMES-assisted standing model from varied seated postures. AB - After spinal cord injury (SCI), intact lower motor neurons can be electrically activated to produce functional muscular contractions and enhance one's capabilities beyond seated activities. Even with neuromuscular electrical stimulation (NMES), significant amounts of hand-support forces are commonly required to move from a sitting to standing position. The goal of this project was to determine initial seated postures that reduce vertical hand-support forces while keeping anterior/posterior hand-support forces below levels that would cause walker slipping or tipping. A multi-segment biomechanical model was further developed and expanded to test multiple combinations of initial postures. The muscles that were analyzed included the vastus lateralis and semimembranosus. Varying the initial knee and hip angles created alternative seated postures. For vastus lateralis stimulation, the lowest vertical hand-support forces (63-66% of body weight) were predicted at the lower (70-74 degrees) and upper (110 degrees) ranges of initial knee flexion. With combined vastus lateralis and semimembranosus stimulation, the lowest predicted vertical hand-support forces were 2-10% of body weight at initial knee flexion angles between 70-82 degrees. Initial hip flexion angles above 110 degrees were required to prevent walker slipping and tipping in these cases. The development of hip extensor torque with semimembranosus stimulation was critical in reducing the vertical hand-support forces. One implication is that when training with parallel bars for NMES assisted standing, hand-support forces should be monitored to avoid conditions that would tip a walker. In future models, optimizing the timing sequence for stimulating muscles may produce smoother coordination of joint rotations and further reduce the vertical hand-support forces. PMID- 15133932 TI - Knee loads in the standard and recumbent cycling positions. AB - The recumbent cycling position (RCP) has become increasingly popular in recent years as a mode of exercise and rehabilitation. However, subtle, but important, differences are expected to exist between the RCP and the standard, upright cycling position (SCP). The differences are due primarily to the altered orientation of the rider's lower extremities relative to gravity. In order to determine if one cycling position may be preferential to another for certain types of rehabilitation, knee loads from a planar, inverse-dynamics model were examined. Nineteen recreational cyclists (24.6 +/- 4.2 yrs) were acclimated to recumbent cycling prior to measuring kinetics and kinematics in both a RCP and SCP (90 rpm; 250 W; max hip-to-pedal distance = 105% leg length). Significance was set at p [symbol: see text] 0.01. Lower-extremity kinematics were not different between the two positions, suggesting that muscle, ligament, and supporting structures travel through the same ranges of motion in both forms of cycling. However, the anterior/posterior forces were altered in such a way that the magnitude of the forces tending to displace the tibia anterior relative to the femur were significantly reduced in the RCP, suggesting that less load may be placed on the anterior cruciate ligament (ACL) while recumbent cycling. No changes in the tension/compression forces were observed at the knee, suggesting no differences in knee stability resulting from compressive forces between the tibia and femur. These findings indicate that the RCP may be beneficial when attempting to minimize ACL loads while utilizing cycling as an exercise and rehabilitative modality. PMID- 15133933 TI - Motion measurements in the jumping of a mountain bike. AB - Mountain biking is generally a much more extreme sport than road cycling. It exhibits large motions and loadings, and involves complicated interactions between the rider and the bike. A dynamic model of a mountain bike and rider is being developed that requires validation by comparing predicted and actual dynamic performance. This paper reports the first step by acquiring experimental data of a moderately extreme maneuver; a mid speed jump and landing and was undertaken as part of a senior level instrumentation class. Both biomechanics and mechanical engineering studies can benefit from this data. A mountain bike and rider are instrumented for video marker tracking during the jumping maneuver. Positions are numerically differentiated to obtain velocities and accelerations. This motion data is examined for continuity and reasonableness. Future manipulation of this data will provide joint reaction data and center of mass motion for a bicycle/rider model validation. PMID- 15133934 TI - Non-contact strain measurement of biological tissue. AB - Strain measurement in biological material is frequently problematic due to material in-homogeneity, high strain to failure, and the relatively low stiffness of most biological tissue. As an alternative to conventional strain measurement techniques, this study investigates the potential of a non-contact optical system capable of measuring three dimensional surface strain maps. The technique makes use of two CCD cameras and pattern recognition algorithms to track the motion of a random speckle pattern on the specimen and compute the displacement and strain fields. The application of this system on biological material is demonstrated on two tissue types that differ widely in material and surface characteristics (cortical bone and ligamentous tissue). A human tibia was loaded in quasi-static three-point bending and the medial collateral ligament from a human knee joint was loaded in tension. The strain field was computed and analyzed in each case. On the relatively stiff cortical bone, accurate results were obtained wherever high resolution imaging of the speckle pattern was possible. Since wicking and seepage from blood vessels during the test affect the speckle pattern, countermeasures are proposed. On the other hand, stretching of ligaments produced large surface strain discontinuities from tissue unfolding, which caused automated pattern recognition algorithms to fail. PMID- 15133935 TI - A new technique of tissue repair for ophthalmic surgery. AB - Ophthalmic surgery currently utilizes suture materials to repair wounds created during eye operations. Although effective, suture-based techniques can result in complications that further impair the patient's vision, such as retinal detachment and scleral perforation associated with strabismus (eye muscle) surgery. Two techniques currently under development avoid sutures altogether, yielding similar strength results, reduced operating time, and simpler methods of repair. The first of these techniques employs a light-activated scaffold-enhanced protein solder to re-adhere the tissue. The second technique utilizes commercially available bioadhesives that have been scaffold-enhanced to improve their handling characteristics. A comparison of these two techniques is given. Initial tensile strength results show a higher strength of repair when a scaffold is utilized, with significantly less variations within each experimental group. Repairs formed using the scaffold-enhanced cyanoacrylate adhesives were the strongest. The tensile strength of extraocular muscle-to-sclera adhesions was 72% stronger than cyanoacrylate alone (4.2 +/- 0.2 N vs. 2.4 +/- 0.4 N) and 78% stronger than native tissue (2.3 +/- 0.4 N). Sclera-to-sclera adhesions were 60% stronger than adhesions formed with cyanoacrylate alone (3.9 +/- 0.2 N vs. 2.5 +/ 0.4 N), while the tensile strength of extraocular muscle-to-extraocular muscle adhesions were 81% of native extraocular muscle tensile strength (5.6 +/- 0.2 N vs. 6.2 +/- 0.3 N), and 50% stronger than adhesions formed using cyanoacrylate alone (3.6 +/- 0.4 N). The data analysis and resulting conclusions favor the less invasive adhesive technique as an alternative for tissue reattachment during ophthalmic procedures. Future experiments will examine the optimization of application parameters and detail tensile strength time course studies. PMID- 15133936 TI - Brown widow (Latrodectus geometricus) major ampullate silk protein and its material properties. AB - Major ampullate (dragline) silk is the main web component as well as the silk that spiders use for a lifeline when they fall. This silk has a breaking stress of 4.6 GPa, which is similar to that of Kevlar. The majority of the previous mechanical testing studies involved the major ampullate silk from orb-weaving spiders. To date, there have been no reports on dragline silk mechanical properties from a cob-weaver, brown widow Latrodectus geometricus. L. geometricus dragline was found to be composed of MaSp1, MaSp2, and MaSp-like proteins all of which have highly conserved amino acid motifs, especially the GGX, GA and poly A for MaSp1 and GPGGX and poly A for MaSp2. These sequences are the same as those found in the silks of orb-weaving spiders. To determine if protein sequences influence the material properties of the silk, mechanical testing was performed on single strands of silk fibers from adult female L. geometricus spiders. The 3 cm long silk fibers were tested for breaking stress and strain with a MTS Synergie 100 mechanical testing system using a 50 g load cell with the cross-head speed set at 10 mm/min. The breaking stress and strain were measured for 20 replicate samples and averaged. The values of 0.83 +/- 0.19 GPa for stress and 0.14 +/- 0.06 for strain shows that brown widow dragline is weaker than the orb weaving spiders. PMID- 15133937 TI - Structural testing of a juvenile prosthetic foot pylon. AB - Clinicians at the Shriner's Hospitals for Children have noticed unexpected catastrophic failures in the 2024-T3 aluminum prosthetic pylons of their juvenile patients. They postulated that small "dings", caused by sharp impacts with playground equipment or other obstacles, might be causing stress concentrations in the pylons. To investigate this, students at the United States Air Force Academy performed a series of tests to determine potential failure modes for the pylon. Initially, a technique for introducing small reproducible "dings" was established. These damaged specimens were then subjected to compressive axial tests to failure, axial fatigue tests (5-100 lbs at 5 Hz), and finally four point bending fatigue tests (370 in-lbs at 5 Hz). The maximum compressive load of the damaged specimen was virtually identical to that of the control specimen (14 kips), and the axial fatigue test was halted with no damage after one million cycles. Two bending fatigue tests resulted in cracks and eventual failure at the load application points of the fixture on the pylon. This has caused us to speculate that failure of the pylons is not due to the dings, but may be due to over-tightening of the pylon attachment points to the foot and the deficient limb. Further testing will be conducted to examine this possibility. PMID- 15133939 TI - A new flexible automated system for the study of exhaled gases. AB - Expired gas analysis has been largely relegated to the measurement of VO2 and VCO2 by a variety of methods. We designed and built a new flexible automated expired gas analysis instrument, the Volume Accumulating Metabolic Monitor (VAMM), capable of simultaneous and continuous quantitative expired gas analysis for a multiplicity of gas species. All expired gas is collected into one of two twin reservoirs. This approach allows analysis of one reservoir while collection occurs into the other. The instrument mixes the expired gas and determines the volume of the desired gas species using a combination of indicator gas dilution and mass spectrometry. We tested the VAMM's ability to measure the 13CO2 2-hr collection after intravenous 13C-aminopyrine. Aminopyrine is metabolized to CO2. Ten healthy volunteers underwent expired gas collection and analysis for 140 min. All studies were performed in the recumbent posture after an 8 hr fast. The initial 20 min were used to establish a baseline 13CO2 production. Following the injection of 2 mg/kg 13C-aminopyrine, expired gas was collected for an additional 120 min. The mean 2-hr 13CO2 enrichment was 7.1 ?2.9 ml (range 3.5 ml to 13.2 ml). This represented a percent collection of 26.8 ?9.1 (range 16.5 to 48.6). Peak excretion occurred at 84.9 ?17.5 min (range 50 min to 108 min). The VAMM provided significantly better 13CO2 excretion profiles than previously described. This technology can easily be customized to study not only other similar metabolic processes but also other gas excretions. PMID- 15133938 TI - A Java mobile phone-based "Home Helper" care report creation support system. AB - In Japan, Home Helpers are employed by hospitals, care companies or the welfare office and are sent to the homes of elderly persons to provide home welfare and care services. They are required to input their reports into the computer at their central office after each care visit. We developed a new Java mobile phone based Home Helper care report creation support system for reducing the data entry time and the communication costs of their reports. The system consists of a Java mobile phone for each Home Helper and a server computer at their office. The server computer sends a Java care report creation support program to the Java mobile phone. The program runs on the mobile phone and supports the report creation in a short time. Therefore, the developed system enables easy report entry by the Home Helper and saves a significant amount of time and communication costs. PMID- 15133940 TI - Analysis of a thermal method for assessing endothelial dysfunction. AB - The presence of atherosclerosis not only affects the normal functioning of the coronary blood vessels but also of the peripheral vasculature. Property measurements made in the peripheral vasculature hence do reflect the condition of the coronary blood vessels. The endothelial cells form the inner lining of the blood vessels, and are responsible for the release of nitric oxide (NO) in order to control the vascular tone. Under normal conditions the artery will dilate in response to increased blood flow, mediated by the release of NO. The hampering of this normal response, caused by certain cardiovascular diseases, is referred to as endothelial dysfunction (EDF). Occlusion of the arm using a standard blood pressure cuff for five minutes followed by sudden release of the occlusion is known to create a reactive hyperemia in a normally functioning vasculature. We propose to measure the EDF by attempting to create this reactive hyperemia in the arm and measuring the temperature response in the hand and forearm, using a computer-based data acquisition system. The rate of temperature fall during occlusion and the temperature rate of rise after release are combined to assess EDF. An engineering analysis of the instrument was performed. Initial studies on normal subjects have indicated that the rate of rise is significantly higher than the rate of fall of temperature. PMID- 15133941 TI - Low dose administration of macrophage colony-stimulating factor in mice. AB - Macrophage-Colony Stimulating Factor (M-CSF) is critical for osteoclast differentiation and development. It has been previously observed that M-CSF administration and over-expression in mice causes an increase in cortical bone formation. We hypothesize that M-CSF increases osteoblast activity indirectly via coupling of these two bone cells. This study examined the impact on bone properties of relatively low doses of M-CSF in mice. Four groups of seven-week old C57BL/6J mice were used: (1) baseline controls, (2) placebo controls, (3) 10 micrograms/kg/day M-CSF, (4) 100 micrograms/kg/day M-CSF. Injections were administered daily for the 21-day study. Three bone labels of calcein and tetracycline were alternately administrated (days 0, 9 and 18) to allow quantification of new bone formation. MicroCT scans (15 micron resolution) were performed on the proximal end of the right tibiae (1.0 mm section of trabecular bone) and left femur mid-diaphysis (0.25 mm cortical section). Dry mass, mineral content and percent mineral composition were obtained from the left tibiae. Functional changes were not detected in the bones of the mice receiving low doses of M-CSF. In particular, as previous studies have reported in mice receiving high doses of M-CSF or transgenic mice overexpressing bone specific M-CSF, changes to cortical bone did not occur with the lower doses. This may indicate that high doses of M-CSF and/or longer periods of administration may be required to observe the anabolic effect of M-CSF on mouse cortical bone. PMID- 15133942 TI - Stimulation of osteogenesis by means of sustained delivery of various natural androgenic hormones. AB - Sex steroids play an essential role in the maintenance of bone health throughout life, and the mechanisms by which these effects are mediated in a subject of much controversy. Osteoblast cells appear to be stimulated by androgens in vitro, however their use in vivo is limited due to the virilizing side effects as well as alterations in the lipoprotein profiles. The use of targeted sustained release of anabolic steroids may stimulate fracture healing without untoward side effects. The specific aims were: (1) to compare fracture healing in a rat femoral defect model using tricalcium phosphate lysine (TCPL) drug delivery systems to deliver T, DHT and AED for long duration; (2) to quantify the level of steroid delivered from the system; and (3) to use bone histomorphometric techniques to analyze new bone formation at the defect site. A total of 125 adult male Sprague Dawley were obtained and acclimatized for two weeks in the animal care prior surgical procedures. All animals were kept on a 12-hour day/night cycle and were fed Purina rodent chow and water ad libitum. The animals were randomly divided into five equal groups (n = 25 per group). Group 1 animals were used as the intact control. Group 2-5 animals were placed under anesthesia and a standard approach was used to create a 6-mm defect using a dental burr in the midshaft of the femur. Group 2 animals were implanted with a sham TCPL delivery system adjacent to the defect. Animals in groups 3, 4, and 5 received a TCPL delivery system loaded with T, DHT, and AED, respectively. Animals were weighed, x-rayed, and blood samples were drawn on a weekly basis. The rats were sacrificed after 3, 6, 9 12 and 15 weeks and reproductive, vital organs, and fracture calluses were collected and analyzed. Morphometric analysis of the femurs revealed that the use of sustained delivery of DHT induced remarkable bone ingrowth compared to the sham and other experimental groups. All treated femurs appeared healthy and normal bone architecture was observed by the end of the 6 week phase. Measurements of the inner perimeter of the bone on the endosteal side showed significant reduction in the androgens treated animals. The quantitative findings confirms our preliminary studies and endorsing the previous data that the sustained delivery of T or its metabolite (DHT) can stimulate the osteoblastic activities in which eventually causes an increase in the cortical bone density. PMID- 15133943 TI - Spaceflight and hindlimb suspension disuse models in mice. AB - Osteoporosis is a disease characterized by low bone mass and structural deterioration of bone tissue, leading to bone fragility and increased susceptibility to fractures. The microgravity of space creates an extreme environment that provides a model for osteoporosis in humans. This greatly accelerated form of osteopenia results in a 0.5-2% loss of bone mass per month. Rat models for this osteoporosis have been examined on many occasions, but STS 108 was the first Space Shuttle flight to use mice. Data reported to date indicate that spaceflight experiments with mice hold promise in predicting some spaceflight effects on humans. Due to the cost and infrequency of flights, ground based models have been developed to mimic the deleterious effects of the microgravity environment. Hindlimb suspension is one such localized model. This model removes gravitational loading from the hindlimbs by suspending the animal by its tail to a guy wire that runs lengthwise across the cage. Because mice had not flown before STS-108, a direct comparison of this model's ability to predict spaceflight results has not been examined. The objective of this research is to closely repeat the STS-108 profile, with hindlimb suspension replacing spaceflight. This includes examining the ability of the protein osteoprotegerin, an osteoclast-inhibiting therapeutic, to mitigate the deleterious effects of skeletal unloading. It is expected that the results will lead to better understanding of the mechanisms of mineralization and bone remodeling to aid in development of countermeasures to prevent spaceflight induced osteoporosis and aid the treatment of osteoporosis here on earth. PMID- 15133944 TI - Effects of sustained delivery of thymoqiunone on bone healing of male rats. AB - The use of natural products as an alternative to conventional treatment in healing and treatment of various diseases has been on the rise in the last few decades. Nigella sativa, a natural herb has long been used as a natural medicine for treatment of many acute, as well as, chronic conditions. These include diabetes, hypertension and dermatological conditions. The specific objectives of this study were: (1) to successfully deliver the active component of black seed called Thymoqiunone (TMQ) at sustained level using TCPL drug delivery system; (2) to evaluate the physiological responses associated with sustained delivery of TMQ in femoral defect animal model (bone healing). A total of 15 adult male rats were randomly divided into three equal groups. Animals in group I served as controls, animals in group II served as sham while animals in group III served as experimental group (femur defect model). Group III animals were surgically implanted with TCPL capsule loaded with 0.02 grams of TMQ and 200 mg vancomycin. Blood samples, x-rays and body weights were collected and recorded weekly. At the end of 30 days post treatment, all animals were sacrificed and vital as well as reproductive organs were collected and analyzed histopathologically. Metabolic biochemical markers were also evaluated. The results of this study revealed the following: (i) gross anatomical observation indicated difference in healing pattern of animals in group III compared to those in group II (sham); (ii) no significant differences in all levels of cholesterol, proteins, malondialdehyde and alkaline phosphatase in all groups; (iii) no significant differences in the wet weights of vital as well as reproductive organs in all the groups. In conclusion, it appears that sustained levels of TMQ can enhance bone healing with little or no side effects on major vital and reproductive organs. PMID- 15133945 TI - Synthesis and evaluation of scaffolds prepared from chitosan fibers for potential use in cartilage tissue engineering. AB - Tissue engineering concepts and methodologies that employ biocompatible matrices or scaffolds have the potential to meet needs encountered in the repair of defects in articular cartilage. A desirable design parameter in the tissue engineering of cartilage in vitro is the development of seeded scaffolds with appropriate structure, composition, mechanical properties and durability that are similar to normal articular cartilage. Previous methods that have used freeze drying and lyophilization techniques to make foams and hydrogels have not met the scaffold characteristics (porosity, compressive elastic modulus, permeability and viscoelastic properties), which are required of scaffolds slated for use in cartilage tissue engineering applications. Thus there is an impetus to design and develop biomimetic scaffolds that mimic the native ECM of articular cartilage, and distribute strain in a bioresponsive manner to signal seeded chondrocytes to synthesize and organize ECM to result in material properties that are in range of natural cartilage. We have employed the method of electrospinning to prepare scaffolds with oriented and random fiber alignment. PMID- 15133946 TI - Effect of hydrogen peroxide on proliferation, apoptosis and interleukin-2 production of Jurkat T cells. AB - Hydrogen peroxide (H2O2) is well known as a cell damaging agent that is produced during normal cell metabolism of aerobic organisms. An excessive production of oxygen metabolites such as H2O2 leads to oxidative stress and disease. On the other hand, it recently was discovered that H2O2 is not only a deleterious oxidant for cells but can also play an important role as a beneficial signaling molecule in certain cells such as T lymphocytes. T lymphocytes are major regulatory cells in the inflammatory cascade and can act by releasing either toxins or beneficial signaling molecules. Understanding how to regulate the actions of H2O2 in T cells will allow for the creation of novel ways to improve the treatment of inflammatory diseases. The current study presents baseline information on the effects of H2O2 on Jurkat cells, a T cell line that we use as a T cell model for therapeutic research. We first determined the half-life of 0 80 mumolar H2O2 added to Jurkat cultures using a realtime H2O2 monitoring system. We then exposed Jurkat cells to such H2O2 concentrations and found that 50 +/- 10 mumolar H2O2 promoted interleukin-2 production in cells activated with anti-CD3 at the T cell receptor plus phorbol myristate acetate as a co-stimulatory signal. These effects were not seen in non-activated, normal Jurkat cells, where H2O2 inhibited cell proliferation and induced apoptosis in a dose-dependent way without affecting interleukin-2 production. Our data indicate that Jurkat cells can model both healthy and inflammatory T cells that respond differently to oxidative metabolites such as H2O2. PMID- 15133947 TI - Comparison of different fabrication techniques used for processing 3-dimensional, porous, biodegradable scaffolds from modified starch for bone tissue engineering. AB - 3 dimensional, porous, biodegradable scaffolds were fabricated using modified starch of varying degree of substitution (DS) by extrusion processing. Freeze drying/lyophilization was also employed to fabricate scaffolds from modified starch. The research efforts have been focused on the comparison of the above mentioned techniques by comparing the properties of the fabricated scaffolds in the paradigm of bone tissue engineering. The physicomechanical properties like porosity, compressive strength and modulus, pore size and microstructure were tested and analyzed by liquid replacement, mechanical testing and scanning electron microscopy respectively. The biodegradability of scaffolds was evaluated by soaking the samples in aqueous medium and Hank's balanced salt solution at 37 degree invitro. The cytotoxicity studies on these scaffolds were also conducted. The scaffolds have a 3D structure consisting of interconnected pores with good porosity, pore size, adequate compressive strength and modulus and exhibit good biodegradability as well as biocompatibility. After further optimization in the processing conditions and parameters they could be made useful for bone tissue engineering. PMID- 15133948 TI - Design and synthesis of a new polymer drug delivery conjugate. AB - This work describes the design and synthesis of a novel polymer conjugate to facilitate intracellular release of attached cargo. Water soluble poly[N(2 hydroxypropyl) methacrylamide] with derivatizable pendant side chains was used as the polymer carrier. The membrane active Tat peptide was masked with poly(ethylene glycol) to inhibit non-specific interactions with exterior cell membranes, and then covalently linked to poly[N-(2-hydroxypropyl) methacrylamide]. A novel heterobifunctional crosslinker was synthesized containing both maleimide and aldehyde functionality to allow attachment of peptides to the polymer backbone through a pH sensitive bond. Analysis of the proposed conjugate by gel permeation chromatography, nuclear absorbance spectroscopy, and absorbance measurements indicate that the desired compound has been synthesized. PMID- 15133950 TI - Design and testing of a fluorescence glucose sensor which incorporates a bioinductive material. AB - One main hurdle in the development of implantable biosensors is that their lifetime is limited by scar tissue formation around the implant. One way to ameliorate this issue would be to use a bioinductive coating to allow normal tissue ingrowth around the sensor. Here, we report design, development and in vitro testing of a prototype fluorescence-based glucose sensor that incorporates a bioinductive material at its tip. Glucose is sensed via a fluorescence-based assay system (Amplex reagent) which is confined to a small chamber separated from the bulk glucose solution by a semipermeable membrane. Excitation is provided by a 530 nm laser, while the emitted light is detected by a photomultiplier tube. In vitro testing of this prototype was done in the presence and absence of a bioinductive material covering the membrane at the sensor/solution interface. In response to a step change in glucose concentration, the output of the sensor increased linearly over time due to accumulation of fluorescent marker molecules as glucose diffused into the recording chamber. The slope of this response increased linearly with increasing glucose concentration, with a sensitivity if 2.1 x 10(-4) V/min per ml glucose/dl solution. The presence of the bioinductive layer did not alter the function of the sensor at the lowest glucose concentrations tested, although responses to higher concentrations saturated, presumably because of depletion of the Amplex reagent within the chamber. In summary, we report that the use of a bioinductive material in an implantable biosensor does not appreciably alter sensor function. PMID- 15133949 TI - A model study of capacitive micromachined ultrasonic transducers fabricated using atomic layer deposition process. AB - In this paper, we present the model study of capacitive micromachined ultrasonic transducers (CMUTs) fabricated by atomic layer deposition (ALD) technology, which uses a self-limiting binary reaction process to produce ultra-thin membranes. Advantages of ALD include precise control of membrane thickness, lower cost due to a reduction in the number of fabrication steps, the potential to use a large variety of materials, and increased reliability due to the enhanced surface quality of the membranes. These capabilities promise fabrication of transducers with superior operating characteristics. However, no study has yet documented sensitivity and power requirements for CMUTs created using ALD. We present here a first-order mechanical and equivalent circuit analysis along with a fabrication process to create and characterize CMUTs using ALD. Simulation results show that these systems have the potential for excellent sensitivity and decreased power requirements. Work to test the fabricated elements is currently underway. PMID- 15133951 TI - Electric field penetration depth of myocardial surface catheters and the measurement of myocardial resistivity. AB - The in vivo measurement of cardiac conductance in mice offers a method to generate an instantaneous left ventricular volume signal. In order to translate the measured conductance to volume, it is necessary to determine the resistivity of the myocardial tissue. This is done using tetrapolar surface conductance catheters, placed on the surface of the left ventricle. It is important to determine the depth of penetration of the electric field of this surface catheter to ensure that the electric field is confined within the myocardium and does not extend into the left ventricle blood volume. The depth of penetration is experimentally determined by a technique described by Foster et al. [2]. PMID- 15133952 TI - Quantification methodology for peripheral quantitative computed tomography (PQCT) data using public domain software. AB - OBJECTIVE: To develop a method for viewing, processing and acquiring 3 dimensional volumetric data from existing ovine spinal peripheral quantitative computed tomography (PQCT) scans of posterior lateral fusion. DESIGN: An image processing development study. BACKGROUND: Existing medical image viewing software can be expensive and difficult to adapt to meet specific research needs. The goals of this study were to produce volume rendering of PQCT scans through processing, masking, and segmentation using public domain software with established source code. METHODS: Raw data files (DICOM format) of 32 PQCT scans from animals receiving spine fusion were obtained. Metal hardware was removed from the images by masks and image segmentation. Calculation of bone macro architecture volumetric data was performed on right and left sides of spines to quantify fusion volume in normalized segments between transverse processes. RESULTS: Images were acquired and opened. Application of image processing techniques made it possible to remove surgical hardware from original images with minimal loss of original PQCT data. Volumes were calculated and normalized to gray-scale of total bone throughout individual selected segments. CONCLUSION: Using public domain software is a cost effective means to view, process, and manipulate PQCT data. Bone macro-architecture can provide quantitative volumetric contributions to ascertain the role of structure on mechanical function. PMID- 15133953 TI - Numerical investigations of intraluminal impedance. AB - The measurement of the electrical impedance inside the esophagus provides information about its status, and is used in the study of the gastroesophageal reflux. This paper presents numerical modeling of the forward problem of the catheter inside the esophagus. The results of the numerical solution for a simple geometry is compared against solution formulated from Green's function approach. This paper also presents investigation of the effect of the esophagus status on the impedance, and the correlation between various factors that affect the impedance. The results of this paper provides a way of relating the esophagus condition to the impedance waveforms as well as in the interpretation of the resulting impedance signals. PMID- 15133954 TI - A new mathematical approach based on orthogonal operators for the detection of interictal spikes in epileptogenic data. AB - This study focuses on the design of orthogonal operators based on unique Electroencephalograph (EEG) signal decompositions in order to detect interictal spikes that characterize epileptic seizures in EEG data. The merits of the algorithm are: (a) in elaborating a unique analysis scheme that scrutinizes EEG data through orthogonal operators designed to extract features that best characterize spikes in epileptogenic EEG data; and (b) in establishing mathematical derivations that provide quantitative measures through the designed operators, and characterize and locate the event of an interictal spike. The uniqueness of this algorithm is in its good performance and simplicity of implementation. Clinical experiments involved 31 patients with focal epilepsy. EEG data collected from 10 of these patients were used initially in a training phase to ascertain the reliability of the observable and formulated features that were used in the spike detection process. Spikes were annotated independently by three EEG experts. On evaluation of the algorithm using the 21 remaining patients in the testing phase revealed a Precision (Positive Predictive Value) of 92% and a Sensitivity of 82%. Based on the 20 to 30-minute epochs of continuous EEG recording per subject, the false detection (FD) rate is estimated at 1.8 FD per hour of recorded EEG. These are good results that support further development of this algorithm for EEG diagnosis. PMID- 15133955 TI - An optimization approach to recognition of epileptogenic data using neural networks with simplified input layers. AB - This study introduces a simplified approach for the implementation of artificial neural networks (ANN) for the recognition of epileptic data in electroencephalograph (EEG) recordings. The training set construction is based on a trend-adaptive polygon which simplifies the search process as it reduces the size of the training set. This data reduction, at a sampling rate of 200 Hz, yielded a reduction ratio of 34% as a minimum to an 81% in the best case scenario. With a higher sampling rate of 500 Hz, a reduction ratio of 73% as a minimum to an impressive 92% in the best case scenario was achieved. The outcome is thus a computationally attractive classifier with a simpler design implementation and with higher prospects for accurate diagnosis. The algorithm was trained and tested with EEG data from four epileptic patients using the k fold cross-validation technique. PMID- 15133956 TI - An integrated auditory-comprehension process augmented through topographical maps and a new eigensystem study. AB - The algorithm developed in this study integrates a frequency analysis of key frequency bands (Alpha, Beta, Delta, and Theta) with the principal component analysis (PCA) in order to validate brain functional mappings associated with the characterization effects of an Auditory/Comprehension task. This study provides added insight to earlier findings involving the Wernicke and Broca's brain areas in relation to language comprehension. A thorough examination of the electroencephalograph (EEG) recordings through the PCA reveals that eigenvectors associated with the largest eigenvalues produce an interesting activity pattern directly attributable to those characteristic behaviors found in the Alpha, Beta, Delta, and Theta frequency bands. The clinical EEG data involved 9 patients at Miami Children's Hospital using the Electrical Source Imaging system with 256 electrodes. An evaluation of spectral arrays is performed using topographic maps of the induced brain activities during both listening and answering phases. This evaluation is then augmented with quantifying measures using the PCA while results are validated through integration of EEG and PCA modalities. Such a representation allows us to bring new insight out on how different patients react under different circumstances, and be able to detect consequently the presence of potential neurological disorders by assessing similar/dissimilar behaviors with respects to all former patients already included in the database. The good results obtained are foreseen to extend the algorithm's application to other brain functional mapping tasks. PMID- 15133958 TI - Dynamic three-dimensional reconstruction and modeling of cardiovascular anatomy in children with congenital heart disease using biplane angiography. AB - Modeling and simulation of cardiovascular biomechanics and fluid dynamics from patient-specific data is a continuing topic of research investigation. Several methodologies utilizing CT, MRI and ultrasound to re-create the three-dimensional anatomy of the cardiovascular system have been examined. Adaptation of these models to pediatric applications has not been studied as extensively. There is significant need for such techniques in pediatric congenital heart disease since local anatomy may exhibit highly unusual geometry, and three-dimensional information would be of significant use for surgical and interventional planning, biomechanical and fluid dynamic simulation, and patient counseling. We report here on the adaptation and application of a three-dimensional reconstruction technique that utilizes bi-plane angiographic images as the base data sets. The method has been validated in a variety of adult imaging situations including coronary artery imaging and intervention. The method uses a skeletonization approach whereby local centerline, diameter, branching and tortuosity of the vasculature are obtained to create the three-dimensional model. Ten patients with a variety of etiology were imaged and 3D reconstructions were obtained. Excellent images were obtained of complex anatomy including the highly branched pulmonary vasculature and Fontan surgical connections. The data were then translated into solid and surface models to facilitate viewing, export into computational fluid dynamic grids, and into files suitable for stereo lithography fabrication (STL). This method appears promising for the dynamic study of complex cardiovascular anatomy found in congenital heart disease. Optimization of the method to facilitate on-line reconstruction and simulation are currently ongoing. PMID- 15133957 TI - Modeling 3-D compliant blood flow with FOSLS. AB - Blood flow in large vessels is typically modeled using the Navier-Stokes equations for the fluid domain and elasticity equations for the vessel wall. As the wall deforms, additional complications are introduced because the shape of the fluid domain changes, necessitating the use of a re-mapping or re-griding process for the fluid region. Typically, this system (fluid, solid, mapping) is solved using an iterative approach in which the fluid, elastic, and mapping equations are solved in series until the iterations converge. We present a new approach based on multilevel minimization of the finite element approximation error using a least-squares (LS) norm. This approach allows for minimization of the error for the entire system or in selected parts. The multilevel LS approach overcomes many shortcomings of standard techniques. Most notably, the computational cost of solving the problem increases linearly with the degrees of freedom and the associated least-squares functional provides an a posteriori error measure. This paper compares the LS finite element approach to other popular numerical methods, specifically, the commercial package CFD-ACE. The focus of the comparison is on accuracy, computational cost, scalability (both parallel and serial), and flexibility. We show that the multilevel LS finite element approach scales optimally (i.e., linearly in serial environments), while the other methods degrade substantially as the problem size increases. PMID- 15133959 TI - Designing blood oxygenators using analogue fluids. AB - Mass transfer and friction factor correlations for commercially available hollow fibre and flat sheet blood oxygenators have been determined experimentally. Water was used as a substitute for blood. The diffusion of oxygen into and out of water has been studied. Three different flow configurations have been investigated: flow inside hollow fibres, flow outside and across woven hollow fibre bundles and flow in thin channels. For flow inside the fibres and in thin channels, the results obtained here are in close agreement with analogous theoretical correlations. The results for flow across bundles of woven hollow fibres may be compared to analogous results for flow in cross flow heat exchangers. PMID- 15133960 TI - EIT reconstructions and Faddeev solutions for a numerically simulated phantom chest. AB - Electrical impedance tomography (EIT) uses measurements of electromagnetic fields to reconstruct the conductivity distribution inside of a body. As a medical imaging technique, EIT has many applications including cardio-pulmonary imaging, cranial imaging, and breast cancer detection. The D-bar method uses inverse scattering techniques and is based on a uniqueness proof in 2-D by Nachman [Ann. of Math.143 (1996)]. In this paper, the solution of the D-bar equation is tested on a model consisting of a numerical simulation of a human chest containing simulated skin, lungs and heart. Faddeev solutions are computed for this simulation and are compared to each other and their known asymptotic behavior. PMID- 15133961 TI - High speed photographic verification of intravascular stent strains during accelerated durability testing. AB - Most stent durability testing does not predict resulting clinical failures. Standards committees have taken on the task of generating more reliable test protocols. Currently, the AAMI/ISO Committee has chosen an experimental direction that allows an investigator to test at ultra high frequencies (above 1000 bpm) as long as the actual strain of the stents can be verified to be moving the same amount per cycle that they do during real time testing (70 to 72 bpm). We have developed a series of techniques and protocols utilizing high-speed photographic analysis to accomplish this. The first aspect requires the utilization of ultra clear silicone mock arteries. It is important that optical anomalies do not cause aberrations in the analysis. Next is the relationship between the angle of the incident lighting and the positioning of the camera in front of the pulsing tube. Finally, a unique set of marking techniques which are used as tracking locations for the automatic measurement systems located in the software of the camera have certain characteristics that are important with respect to accurate measurements. In each case the particular setup must have independent verification of the accuracy of the measurements. We will review these variables and demonstrate how this system can be utilized to verify strains of the inside wall, outside wall, and stent. PMID- 15133964 TI - Novel hyaluronan esters for biomedical applications. AB - A series of novel hyaluronan (HA) esters with aliphatic chains of different length were synthesized. A silylated complex of HA with cetyltrimethylammonium cations (silyl HA-CTA) was used as the starting material. The reactions were performed in xylenes or no solvent with acid chlorides as the acylation agents. The disappearance of all characteristic FT-IR vibration bands associated with the -OSi(CH3)3 groups and the appearance of the strong ester carbonyl peak at 1753 cm 1 demonstrated the success of esterification. Thermoplasticity was achieved with HA esters of high aliphatic acids. It was found that the longer the ester chain, the lower the melting points, so different melting temperatures can be obtained by adjusting the acid chloride chain length to meet various needs. PMID- 15133963 TI - Probing the elastic nature of spider silk in pursuit of the next designer fiber. AB - Spider silk, one of nature's greatest accomplishments, has a combination of strength and elasticity that is unrivaled. Spiders produce up to 7 different silks; each one with a unique combination of tensile strength and elasticity that allows the spiders' web to hold prey while being resilient enough not to break upon impact. In an attempt to determine the sequences responsible for these enviable mechanical properties, several different amino acid motifs have been defined. Much of the recent work is now concentrated on correlating amino acid motifs with a specific mechanical property. The current hypothesis is that the strength property of spider silk is conferred by a poly-alanine or alanine rich (An or GAn) motif whereas the elastic nature of spider silk is conferred by the amino acid motif, GPGXX, where X is Q, S, A, G, or Y. Despite the fact that these different motifs are now known, the combination of strength and elasticity are yet to be duplicated ex vivo. In an attempt to verify that the GPGXX motif imparts elasticity to the spider silk, the number of repeats and/or the amino acid composition of the Argiope aurantia "elastic motif" were varied and expressed in various strains of E. coli to change the elastic properties of the resulting film and/or fiber. Concurrent with work on the elasticity motif is ongoing work on the strength module. Understanding these two different motifs will aide efforts to produce a designer biomaterial for medical, commercial, and military applications. PMID- 15133962 TI - CFD simulation of centrifugal cells washers. AB - The feasibility of using computational fluid dynamics to guide the design of better centrifuges for processing shed blood is explored here. The velocity field and the rate of protein removal from the shed blood have been studied. The results indicate that computational fluid dynamics could help screen preliminary centrifuge bowl designs thus reducing the number of initial experimental tests required when developing new centrifuge bowls. Though the focus of this work is on washing shed blood the methods developed here are applicable to the design of centrifuge bowls for other blood processing applications. PMID- 15133965 TI - Toward immobilized antibody microarray optimization: print buffer and storage condition comparisons on performance. AB - Performance issues in protein arrays differ substantially from DNA microarrays because of the different intrinsic construction and stability of proteins versus DNA, and the impact on biological recognition phenomena required for assay from a surface-immobilized state. Maintenance of the capture bioactivity and performance of antibodies on microarray substrates for relatively long periods (under storage or shipping) requires carefully designed and optimized print buffer and storage conditions. We have focused on selection of several additives added to antibody print buffers with different concentrations, and storage of printed antibody slides under different temperatures from 1 day to 4 weeks. Activities of these slides were compared in capture assay formats in order to compare the effects of print buffer and storage conditions of antibody microarrays. Commercial polymer coated Optarray slides exhibited better performance over Codelink slides in terms of activity of anti-human TNF alpha and IL-1 beta. IL-1 beta antibodies immobilized on both commercial surfaces showed much higher activity than TNF alpha antibodies, regardless of source or vendor. Addition of 1% (v/v) PEO to the print buffer increased the printed analyte capture activity of IL-1 beta on Optarray slides compared to standard print buffer without any additives. This helps to maintain antibody immobilized analyte capture activity for 4 weeks when stored at -70 degrees C. PMID- 15133966 TI - Constitutive models for a poly(e-caprolactone) scaffold. AB - We investigate material models for a porous, polymeric scaffold used for bone. The material was made by co-extruding poly(e-caprolactone) (PCL), a biodegradable polyester, and poly(ethylene oxide) (PEO). The water soluble PEO was removed resulting in a porous scaffold. The stress-strain curve in compression was fit with a phenomenological model in hyperbolic form. This material model will be useful for designers for quasi-static analysis as it provides a simple form that can easily be used in finite element models. The ASTM D-1621 standard recommends using a secant modulus based on 10% strain. The resulting modulus has a smaller scatter in its value compared to the coefficients of the hyperbolic model, and it is therefore easier to compare material processing differences and ensure quality of the scaffold. A third material model was constructed from images of the microstructure. Each pixel of the micrographs was represented with a brick finite element and assigned the Young's modulus of bulk PCL or a value of 0 for a pore. A compressive strain was imposed on the model and the resulting stresses were calculated. The elastic constants of the scaffold were then computed using Hooke's law for a linear-elastic isotropic material. The model was able to predict the small strain Young's modulus measured in the experiments to within one standard deviation. Thus, by knowing the microstructure of the scaffold, its bulk properties can be predicted from the material properties of the constituents. PMID- 15133967 TI - Postmortem retrieved canine THR: femoral and acetabular component interaction. AB - Dogs are the preferred animal model for testing of human total hip replacements (THRs). A postmortem retrieval program for clinical, cemented, canine THR was established to analyze the long-term performance of THRs in dogs and to compare that performance to postmortem retrievals of human THRs. The purpose of the present study was to analyze the interaction between the femoral and acetabular components. Thirty-eight postmortem retrievals from 29 dogs were donated and analyzed. The acetabular components (ACs) were measured for volumetric wear and graded for articulating surface damage. Femoral and acetabular components were mechanically tested for implant stability. Digital image analysis was performed on contact radiographs of transverse femoral slices. Of 14 cases with a firmly implanted femoral component (FC). 6 articulated against loose ACs. Of 24 cases with a loose FC, 16 articulated against loose ACs. Only 4 specimens had both components firmly implanted, and 14 specimens had both components loose. There was a significant positive correlation between AC volumetric wear and FC loosening; however, there was no evidence of osteolysis or wear debris induced osteolysis as seen in human postmortem retrieval studies. There was a significant but weak negative correlation between FC loosening at the cement/bone interface and AC scores reflecting damage to the rim and creep across the entire AC. Although implant-on-implant damage to the AC was expected to positively correlate with FC loosening, this was not found. Researchers need to look at interactions between AC and FC to understand how the failure of one component affects performance of the other. PMID- 15133968 TI - An assessment of equine cartilage degeneration. AB - Millions of Americans suffer from osteoarthritis, a joint disease characterized by cartilage degradation and subchondral bone sclerosis. However, little is known about its pathology. It remains to be discovered which comes first in the progression of osteoarthritis: subchondral bone remodeling or cartilage degeneration. This study assessed equine cartilage degeneration, based upon measurements of cartilage thickness and Indian ink stain uptake. By gaining a greater understanding of the determining factors in cartilage degeneration, we may be able to better understand the pathomechanics of osteoarthritis. In seven horses, joint regions of interest were harvested bilaterally and isolated into planar osteochondral samples using a band saw and stored at -20 degrees C. After thawing, the samples' articular surfaces were dyed using an Indian ink stain to highlight articular cartilage degeneration. Digital images of the samples were taken before and after the staining, and mean pixel values for the pre- and post ink images were measured using image analysis software. Reflectance Score (RS) was calculated using mean pixel values normalized between grayscale calibration standards. Articular cartilage thickness was measured at five random locations on each sample from images taken in the transverse orientation. Statistical analysis found no significant effect of limb side for either RS values or thickness, allowing data from right and left limbs to be grouped for analysis. There is a statistically significant correlation between joint region and thickness, as well as between joint region and RS; however, there was no statistical correlation between thickness and RS. PMID- 15133969 TI - Potential role of proinflammatory cytokines in nerve damage related bone loss. AB - An estimated 375,000 people are currently suffering from spinal cord injuries and another 1.5 million are afflicted by peripheral nerve damage in the United States. Wolf's Law states that a bone grows or remodels in response to the stresses that are placed on it. Forces applied to bones that occur due to normal daily activity allow for healthy resorption and formation of bones. Periods of immobilization caused by nerve damage have a profound effect on the integrity of bone, causing an increased risk of bone fracture. The need for investigating ways of combating this secondary effect of nerve damage is imperative to the long-term health of spinal cord injury and peripheral nerve damage patients. Our lab uses two sciatic nerve damage models in mice to mimic the bone loss caused by recoverable, sciatic nerve crush (NC), and non-recoverable, sciatic neurectomy (NX), injuries. We are examining the hypothesis that recoverable damage actually causes an accelerated loss of bone mass compared to the permanently damaged nerve because of the transport of proinflammatory cytokines from the site of the nerve damage to the locally affected bone. This inflammatory response, and the hypothesized differences between the two models, will be examined via ELISA of the quadriceps to investigate the relative degree of proinflammatory cytokines local to the damage site. Understanding the cellular mechanisms that occur at nerve injury sites will allow for improved care and long-term treatment of patients. A preliminary analysis of the bone loss associate with these two nerve injury models indicate approximately a 50% greater decline in femoral mass of the NC femur compared to the NX limb, supporting the proinflammatory hypothesis. PMID- 15133970 TI - Assessment of subchondral bone mineral density in equine metacarpophalangeal and stifle joints. AB - Functional relationships between articular cartilage and underlying subchondral bone have been shown to be associated with the progression of osteoarthritis (OA). However, quantifiable factors substantiating this relationship do not exist. Therefore, the study objective involved quantifying subchondral bone mineral density (BMD) of the equine metacarpophalangeal (MCP) and stifle joints as a step in determining if regional subchondral BMD may be associated with OA changes. BMD was bilaterally quantified using dual energy xray absorptiometry (DEXA) for four subchondral regions: palmer and dorsal aspect of the medial 3rd metacarpal (MC3P, MC3D), medial femoral condyle (MFC), and the medial trochlear ridge (MTR). BMD (g/cm2) was measured using a 5.2 x 5.2 mm region of interest in the subchondral bone area. To account for the differences in thickness, BMD (g/cm2) was divided by sample thickness providing volumetric BMD (g/cm3). No significant effect of side on BMD values was found (p > 0.72). However, there were significant differences in BMD found between all regions (p < 0.05). The volumetric BMD of the MC3P region was significantly greater than the MC3D and the MC3D was greater than the MFC, while the MFC was greater than the MTR (MC3P > MC3D > MFC > MTR). The MCP regions had a higher volumetric BMD than the stifle regions possibly due to higher weight distribution, smaller articular surface area, and joint geometry. The adaptive bone structural variations between the regions studied were shown to be significant. Mechanical properties of the overlying cartilage are currently being analyzed to correlate with these results and provide a possible diagnostic method to investigate OA progression. PMID- 15133971 TI - Pneumatic strength assessment device: design and isometric measurement. AB - In order to load a muscle optimally during resistance exercise, it should be heavily taxed throughout the entire range of motion for that exercise. However, traditional constant resistance squats only tax the lower-extremity muscles to their limits at the "sticking region" or a critical joint configuration of the exercise cycle. Therefore, a linear motion (Smith) exercise machine was modified with pneumatics and appropriate computer control so that it could be capable of adjusting force to control velocity within a repetition of the squat exercise or other exercise performed with the device. Prior to application of this device in a dynamic squat setting, the maximum voluntary isometric force (MVIF) produced over a spectrum of knee angles is needed. This would reveal the sticking region and overall variation in strength capacity. Five incremental knee angles (90, 110, 130, 150, and 170 degrees, where 180 degrees defined full extension) were examined. After obtaining university-approved informed consent, 12 men and 12 women participated in the study. The knee angle was set, and the pneumatic cylinder was pressurized such that the subject could move the barbell slightly but no more than two-centimeters. The peak pressure exerted over a five-second maximum effort interval was recorded at each knee angle in random order and then repeated. The average of both efforts was then utilized for further analysis. The sticking region occurred consistently at a 90 degrees knee angle, however, the maximum force produced varied between 110 degrees and 170 degrees with the greatest frequency at 150 degrees for both men and women. The percent difference between the maximum and minimum MVIF was 46% for men and 57% for women. PMID- 15133972 TI - Effect of floor slope on submaximal lifting capacity. AB - In order to reduce injuries due to lifting a box from the floor, maximal acceptable weights of lift (MAWL) have been established for a level surface. However, an inclined surface condition may be encountered on a jobsite. The purpose of this investigation was to determine if facing up or down a sloped surface affects MAWL. After obtaining university-approved informed consent, 20 apparently healthy men (age = 22.4 +/- 1.4 yrs) and 20 women (age = 22.0 +/- 1.9 yrs) determined floor to knuckle height MAWL using the psychophysical approach. After a familiarization day, two data collection days were completed with the uphill and level (+20, +10 and 0 degree) or downhill and level (-20, -10 and 0 degree) lifting capacities determined. A cadence of four lifts/min was used after starting with an unknown load that participants adjusted after each lift. No differences (p > 0.05) in level MAWL were found on the downhill day compared to the uphill day. While the men lifted significantly more than the women in every condition (p < 0.001), no differences were found across the lifting conditions (p > 0.05). The men averaged a MAWL of 24.7 kg across the five conditions (average standard deviation (SD) = 7.4 kg), the women averaged 14.8 kg (average SD = 3.1 kg). While these findings would suggest no changes in lifting guidelines for a sloped surface within 20 degrees of level, other factors such as lifting technique and the stress placed on the low-back should be examined to assess risk of injury in these different conditions. PMID- 15133973 TI - Axial and rotational alignment of the leg. AB - The axial and rotational alignments of the lower extremity are commonly referenced independently, with minimal research on whether coexistent axial and rotational malalignment cause pathologies. The present study analyzed whether a correlation exists between the axial and rotational alignments of the leg. The methodology to measure both alignments was adapted for computer tomography. Fifty patients were analyzed at five reference images to determine axial and rotational alignment. The reference images included the femoral head, the femoral shaft (at the level of the lesser trochanter), the distal femur, the proximal tibia, and the ankle joint. Axial alignment was calculated by using horizontal and vertical measurements of the location of the femoral head, the distal femur, and the ankle joint. Rotational alignments of femur, knee, and tibia were calculated using four angles: proximal femoral, distal femoral, proximal tibial, and ankle joint angles defined relative to a fixed reference. Pearson correlation analysis between axial alignment and the three mentioned rotational alignments were calculated. The correlation coefficient values ranged between -0.15-0.07 when comparing axial to rotational alignment, indicating that a week correlation exists between the two alignments. Though these results were derived using highly reproducible methods, the hypothesis of an existing correlation between the axial and rotational alignments of the leg was rejected. These findings allow for an improved understanding of lower extremity mechanics, which merit importance when considering pathologies of the leg and the surgical techniques that could ultimately benefit patients suffering from these pathologies. PMID- 15133974 TI - Comparison of strength properties of normotensive and hypertensive rat pulmonary arteries. AB - A series of tests were conducted to quantify the difference in the mechanical properties of normo- and hypertensive pulmonary arteries. A bubble-test design was employed to measure the biaxial properties of a segment of artery. The test results compare the properties at multiple orientations of the trunk, right, and left pulmonary arteries from normal (Control) and monocrotaline-treated male Long Evans wild rats that ranged in age from 8 to 17 weeks old, along with some preliminary results from hypoxic Long-Evans knock-out rats. Data show little difference between the stress-strain relationship of the control pulmonary arteries and that of the monocrotaline-treated pulmonary arteries. However, the preliminary results from the hypoxic pulmonary arteries show that the arterial material strains less before the onset of strain-stiffening behavior. The longitudinal orientation exhibits strain stiffening at lower strains than does the circumferential orientation. The differences between the left and right main arteries are minor. The trunk consistently demonstrates less stiffening in the region of larger strains for all conditions. PMID- 15133975 TI - Stress and strain in rat pulmonary artery material during a biaxial bubble test. AB - A biaxial bubble test has been designed to ascertain the mechanical properties of rat pulmonary arteries. The analytical procedure used to estimate stress and strain from the resulting test data is presented along with some analytical results. The bubble test was performed by loading a flat piece of rat pulmonary artery into a test fixture beneath a circular opening; the material was subsequently pressurized from below, producing a "bubble" of deformed material. Due to the anisotropy of the rat pulmonary artery, the resulting bubble was ellipsoidal in shape. Test results were recorded in the form of side-view images taken from various angles at incremental values of pressure. Average strains were estimated with the use of image analysis to measure changes in the bubble perimeter during inflation. Formulations for isotropic materials were applied to estimate stresses based on the anisotropic geometry of the bubbles produced during testing; some results of this preliminary analysis are presented here. Results from this analysis show differences in mechanical properties of the rat pulmonary artery from those of healthy versus hypertensive rats. PMID- 15133976 TI - Cardiocirculatory performance at high altitude. AB - The effects of high altitude and physical activity on cardiocirculatory performance are assessed using blood pressure ejection fraction. PMID- 15133977 TI - Linear and non-linear parameters of heart rate variability during static and dynamic exercise in a high-performance dinghy sailor. AB - The analysis of heart rate variability (HRV) is recognized as a powerful non invasive tool to evaluate the influence of autonomic nervous system on the heart and the heart-brain interaction. It is well-known that relatively high HRV is correlated to health condition while low HRV corresponds to cardiovascular disease or could be caused by mental stress, depression or exercise. In order to investigate heart-brain interaction we compare linear and non linear parameters calculated from HRV signals recorded in the same subject at similar heart rate (HR) values, during two different conditions, i.e. static exercise during sailing and dynamic exercise on cycloergometer. In the study, performed in one high performance would class dinghy sailor, the HR was recorded at rest and during the two types of exercise. For the analysis, tachogram tracts with similar HR values were considered. The power spectral densities in very-low, low (LF) and high (HF) frequency bands were evaluated as well as the LF/HF ratio, the two standard deviations (SD1 and SD2) of the Poincare plot, the beta values and the fractal dimension (FD). The results indicate a decrease of HRV, LF, HF, SD1 and SD2 parameters, as well as an increase of beta and FD during both types of exercise compared to rest. However, the higher values of LF, LF/HF ratio and SD2 as well as the lower value of FD in upwind sailing in comparison to dynamic exercise on cycloergometer, at similar HR, suggest a different sympatho-vagal modulation on cardiac function and therefore a different heart-brain interaction in these isometric and isotonic exercises. PMID- 15133979 TI - Simulation of branching blood flows on parallel computers. AB - We present a fully parallel nonlinearly implicit algorithm for the numerical simulation of some branching blood flow problems, which require efficient and robust solver technologies in order to handle the high nonlinearity and the complex geometry. Parallel processing is necessary because of the large number of mesh points needed to accurately discretize the system of differential equations. In this paper we introduce a parallel Newton-Krylov-Schwarz based implicit method, and software for distributed memory parallel computers, for solving the nonlinear algebraic systems arising from a Q2-Q1 finite element discretization of the incompressible Navier-Stokes equations that we use to model the blood flow in the left anterior descending coronary artery. PMID- 15133978 TI - Improving estimation of cardiac vagal tone during spontaneous breathing using a paced breathing calibration. AB - Respiratory sinus arrhythmia (RSA) is a commonly employed non-invasive measure of cardiac vagal control. It has been demonstrated that respiratory parameters such as tidal volume and respiratory frequency can change RSA without altering tonic vagal activity. Thus, within-individual comparisons of cardiac vagal control across different behavioral tasks might benefit from an adjustment for respiratory confounds. We tested an adjustment method using transfer function analysis and paced breathing at 3 different respiratory frequencies as the basis for regressing out respiratory related RSA changes in a task where breathing was not controlled. Electrocardiogram and calibrated respiration were recorded with the LifeShirt system from 15 young adult participants. Time series of RR intervals and lung volume change were computed and the respiration-to-RR-interval transfer-function magnitude (RSA-TF, in ms/liter) estimated. Mean (SD) of RSA-TF was 142 (68) at 9 breaths/min, 78 (52) at 13.5 breaths/min, 57 (43) at 18 breaths/min, and 121 (56) during baseline, with a respiratory frequency of 12.5 (3.8) breaths/min. At baseline, measured and predicted RSA-TF values (mean 94 +/- 82) differed significantly and correlated only moderately (r = 0.67). Factors contributing to a less than perfect correlation included slightly elevated subjective anxiety levels and hyperventilation during paced breathing, both of which may have affected cardiac vagal tone. This study demonstrates a novel procedure for computing a respiratory unrelated RSA index. Results provide some support for the utility of this adjustment method for improving the estimation of cardiac vagal tone from RSA, but also indicate that the paced breathing procedure may need to be further refined. PMID- 15133980 TI - TGF-beta and TNF-a affect cell surface proteoglycan and sialic acid expression on vascular endothelial cells. AB - Atherosclerosis is the formation of plaques in the arterial wall brought about by numerous events including the accumulation of oxidized low density lipoprotein (LDL), stimulation of inflammatory responses, the release of cytokines, and the attachment of monocytes to the arterial wall. Proteoglycans are implicated in many aspects of atherosclerosis including the metabolism of lipoproteins, regulation of cytokine activity, cell adhesion, and modification of the extracellular matrix. Due to their complex role in molecular recognition and cellular adhesion, the glycosaminoglycan (GAG) chains attached to the proteoglycan core and sialic acids on the terminal ends of the glycan chains are of interest. This study investigated the effects of exposure to transforming growth factor-beta 1 (TGF-beta 1) and tumor necrosis factor-a (TNF-a) on the expression of cell surface GAGs and sialic acids on human umbilical vein endothelial cells (HUVECs). Initial results show that TGF-beta 1 affected GAG expression compared to a control condition. Results also show that the combination of TGF-beta 1 and TNF-a affected GAG expression differently than does TGF-beta 1 alone. Additionally, TNF-a decreased the number of sialic acid residues per cell and TGF-beta 1 slightly upregulated sialic acid expression as compared to the control. The combination of the two cytokines showed a larger upward trend in this value. These data indicate that TNF-a and TGF-beta 1 play a role in the expression of GAG chains and sialic acids on the cell surface. Further study may clarify the implications of these findings for atherosclerosis. PMID- 15133981 TI - Kullback-Leibler clustering of continuous wavelet transform measures of heart rate variability. AB - Power spectral analysis of beat-to-beat heart rate variability (HRV) has provided a useful means of understanding the interplay between autonomic and cardiovascular functionality. Despite their utility, commonly employed frequency domain techniques are limited in their prerequisite for stationary signals and their inability to account for temporal changes in the power spectral and/or frequency properties of signals. The purpose of this study is to develop an algorithm that utilizes continuous wavelet transform (CWT) parameters as inputs to a Kohonen self-organizing map (SOM), providing a method of clustering subjects with similar wavelet transform signatures. Continuous interbeat-intervals were recorded (Portapres monitor at 200 Hz) during a perception of affect test in 79 African-American volunteers (ages 21-83), where after a 5-min baseline, participants evaluated emotional expressions in sentences and pictures of faces, followed by a 5-min recovery. Individual HRV biosignals from each session were pre-processed (artifact replacement and signal resampling at 2 Hz) and a CWT was applied (db9 wavelet basis function over 32 scales). Standard deviations of resulting wavelet coefficients at each scale were calculated, normalized, and used as inputs into a SOM with Kullback-Leibler divergence as the dissimilarity measure used for clustering. Differences in subject demographics between two final clusters were assessed via two-independent-groups t-tests or chi-square or Fisher's exact tests of contingency tables. Significant differences were found for age, initial systolic blood pressure, smoking status, and mean s.d. of coefficients in the high frequency band (0.15-0.4 Hz). These findings may have clinical significance and the developed algorithm provides an alternative means of analyzing HRV data originating from populations with complex covariates. PMID- 15133982 TI - Continuous monitoring of cardiac output from TCG signals. AB - Continuous measurement of cardiac output (CO) is an important and difficult measure to obtain in an ambulatory environment. A novel ambulatory monitoring system (LifeShirt, VivoMetrics, Inc., Ventura, CA, USA) with three Inductive Plethysmographic (IP) sensors embedded in a garment, enables continuous monitoring of respiration from the ribcage and abdomen areas, and captures thoracocardiograph (TCG) signals from the thorax at the level of the left ventricle. This TCG signal provides a non-invasive measure of the volumetric contractions of the heart. The raw TCG signal must undergo extensive signal processing and digital filtering to extract a volume curve similar to the ventricular volume curve obtained through echocardiography. Typically the respiratory component has an amplitude of over twenty times that of the stroke volume curve. This investigation compares various signal processing algorithms such as spectral subtraction and adaptive filtering to separate these 2 components, which can occupy the same frequency band. These algorithms make use of the ribcage and abdominal signals to predict the respiratory component within the TCG signal. A dual axis accelerometer that measures posture and levels of activity aids filtering movement artifact. With the addition of a single lead ECG, ensemble averaging is used to smooth artifact in the signal, and CO may be obtained by including a heart rate measure. Additional measures can be derived including left ventricular systolic time intervals such as Pre-ejection period, Peak ejection rate and time to peak ejection rate. The results show that increases and decreases in SV and CO can be measured over time. PMID- 15133983 TI - Study of in vitro mitral valve filling flow. AB - Diastolic function is a good indicator of overall cardiac health. This study is in support of a non-invasive technique to measure diastolic function. Interpretation of mitral flow, and early diagnosis of normality or dysfunction, can be enhanced by improved understanding of the fluid dynamics. A simple in vitro model of mitral flow is being used to illustrate vortex dynamics associated with diastolic inflow. A range of tailored mitral flow waveforms are being developed to mimic normal velocity profiles. Laser Doppler velocimetry (LDV) is being used to measure the flow speed. By combining flow visualization, particle image velocimetry and LDV, vortex ring signatures including size, circulation, and propagation speed are being examined as possible metrics for diastolic dysfunction. The detection of these signatures in the downstream flow will be compared to clinical waveforms derived from Doppler ultrasound. PMID- 15133984 TI - Echo PIV for flow field measurements in vivo. PMID- 15133986 TI - Advantages in using multi-frequency driving ultrasound for optimizing echo particle image velocimetry techniques. AB - We have recently developed an ultrasound based velocimetry technique, termed echo particle image velocimetry (echo PIV). This method takes advantage of the non linear backscatter characteristics of ultrasound contrast microbubbles when exposed to certain ultrasonic field. Preliminary in vitro, animal and clinical studies have shown significant promise of this method for measuring multiple velocity components with good temporal and spatial resolution. However, there is still difficulty in maximizing the non-linearity of bubble backscatter using conventional Gaussian-pulse excitation techniques because significant harmonic components may not be produced at modest pressure amplitudes and the higher incident pressure amplitudes required to induce non-linear behavior may cause bubble destruction. We present here a potential solution to this problem through the use of multi-frequency excitation. A rectangular pulse with multiple harmonics is used to drive the bubble. The backscatter process is studied through a modified Rayleigh-Plesset equation. Results show that the rectangular wave is effective in improving the visibility of microbubbles with ultrasound backscattered efficiency significantly higher than the widely used Gaussian waveform. Use of rectangular pulses with 4 and 2 harmonics showed no significant difference in bubble backscatter behavior, indicating that a two-frequency excitation may be sufficient to induce non-linear behavior of the microbubbles practically at modest incident pressures. PMID- 15133985 TI - Ultrasound wave propagation in tissue and scattering from microbubbles for echo particle image velocimetry technique. AB - Nonlinear wave propagation in tissue can be employed for tissue harmonic imaging, ultrasound surgery, and more effective tissue ablation for high intensity focused ultrasound (HIFU). Wave propagation in soft tissue and scattering from microbubbles (ultrasound contrast agents) are modeled to improve detectability, signal-to-noise ratio, and contrast harmonic imaging used for echo particle image velocimetry (Echo-PIV) technique. The wave motion in nonlinear material (tissue) is studied using KZK-type parabolic evolution equation. This model considers ultrasound beam diffraction, attenuation, and tissue nonlinearity. Time-domain numerical model is based on that originally developed by Lee and Hamilton [J. Acoust. Soc. Am 97:906-917 (1995)] for axi-symmetric acoustic field. The initial acoustic waveform emitted from the transducer is assumed to be a broadband wave modulated by Gaussian envelope. Scattering from microbubbles seeded in the blood stream is characterized. Hence, we compute the pressure field impinges the wall of a coated microbubble; the dynamics of oscillating microbubble can be modeled using Rayleigh-Plesset-type equation. Here, the continuity and the radial momentum equation of encapsulated microbubbles are used to account for the lipid layer surrounding the microbubble. Numerical results show the effects of tissue and microbubble nonlinearities on the propagating pressure wave field. These nonlinearities have a strong influence on the waveform distortion and harmonic generation of the propagating and scattering waves. Results also show that microbubbles have stronger nonlinearity than tissue, and thus improves S/N ratio. These theoretical predictions of wave phenomena provide further understanding of biomedical imaging technique and provide better system design. PMID- 15133987 TI - Biomechanics of seat belt restraint system. AB - Seat belt system restrains and protects occupants in motor vehicle crashes and any slack in seat belt system induces additional loading on occupant. Signs of belt loading are more obvious in high-speed frontal collisions with heavy occupants. However subtle changes may occur at low speeds or with low forces from occupants during rollovers. In certain cases, the seat belt webbing is twisted and loaded by the occupant. The loading of webbing induces an observable fold/crimp on the seat belt. The purpose of the study is to biomechanically evaluate the force required to produce such marks using an anthropometric physical test dummy. Two tests were conducted to determine the amount of force required to put an observable fold/crimp in a shoulder belt. A head form designed by Voight Hodgson was used to represent the neck which interacted with the belt. The force was applied with a pneumatic pull ram (central hydraulic 89182 N) and the force was measured with a 44,000 N transducer load cell (DSM-10K). Results indicate that the force of over 1,000 N produced a fold or crimp in the belt. PMID- 15133988 TI - Multivariate head injury threshold measures for various sized children seated behind vehicle seats in rear impacts. AB - Government recommendations to place children into the rear areas of motor vehicles to avoid airbag induced injuries have been complicated by the fact that most adult occupied front seats will collapse into the rear area during rear impacts, and thus pose another potentially serious injury hazard to rear-seated children. Many variables affect whether or not a front seat occupant will collapse into the rear child, and whether that interaction could be injurious to the child. For instance, the severity of rear impact, coupled with front and rear occupant sizes (mass and stature), and the level of front seat strength, all interrelate to influence whether or not a rear seated child is likely to be impacted and possibly injured. The most common types of child injuries in these instances are head and chest injuries. In this study, a "high-low" experimental method was employed with a multi-level "factorial analysis" technique to study "multivariate" biomechanics of child head injury potential determined from rear seated 3 and 6 year-old child surrogates in different types of vehicle bodies mounted to a sled system. The sled-buck systems were towed rearward into crushable barriers that matched the crash pulses of the vehicle types being tested. Various sizes of adult surrogates (i.e. 50 kg up to 110 kg), seated in both the "typical" low strength "single recliner" collapsing type front seat (i.e. 3.2 kN) and a much stronger "belt-integrated" seat design (i.e. up to 14.5 kN), were tested in the two different "sled body-buck" set-ups at various impact levels (i.e. 22.5 to 50 kph). One set-up used a popular minivan vehicle body with "built-in booster" seats for the 3 year-old. The other used a 4-door family sedan vehicle body with the 6 year-old in a standard rear bench seat. The parameters of the tests enabled the experimental data to be combined into polynomial "head injury" functions of the independent variables so the "likelihood" of rear child head-injury potential could be "mapped" over ranges of the key parameters. Accident cases were compared with predictions to verify the methodology. PMID- 15133989 TI - The locking and unlocking characteristics of a seat belt inertial locking system due to acceleration. AB - An inertial locking mechanism of a seatbelt is designed to prevent further seatbelt extension when it exceeds an acceleration threshold. The acceleration threshold varies with the orientation of the mechanism and direction of acceleration. A setup and procedure were created to test the acceleration threshold of the inertial locking mechanism of a seatbelt in various orientations. For an example of an inertial locking mechanism a 1984 Ford Mustang seatbelt mechanism was used. During testing, the mechanism was put into an orientation that simulated accelerations that could be seen in normal collisions. The mechanism responded by locking with an average acceleration of 2.535 g. The mechanism was then tested in orientations that would simulate those seen by a vehicle in a rollover situation. In certain orientations, the mechanism was locked while at rest, but became unlocked when accelerated. PMID- 15133990 TI - Testing and injury potential analysis of rollovers with narrow object impacts. AB - Recent statistics highlight the significant risk of serious and fatal injuries to occupants involved in rollover collisions due to excessive roof crush. The government has reported that in 2002. Sports Utility Vehicle rollover related fatalities increased by 14% to more than 2400 annually. 61% of all SUV fatalities included rollovers [1]. Rollover crashes rely primarily upon the roof structures to maintain occupant survival space. Frequently these crashes occur off the travel lanes of the roadway and, therefore, can include impacts with various types of narrow objects such as light poles, utility poles and/or trees. A test device and methodology is presented which facilitates dynamic, repeatable rollover impact evaluation of complete vehicle roof structures with such narrow objects. These tests allow for the incorporation of Anthropomorphic Test Dummies (ATDs) which can be instrumented to measure accelerations, forces and moments to evaluate injury potential. High-speed video permits for detailed analysis of occupant kinematics and evaluation of injury causation. Criteria such as restraint performance, injury potential, survival space and the effect of roof crush associated with various types of design alternatives, countermeasures and impact circumstances can also be evaluated. In addition to presentation of the methodology, two representative vehicle crash tests are also reported. Results indicated that the reinforced roof structure significantly reduced the roof deformation compared to the production roof structure. PMID- 15133991 TI - Edge encoding mechanisms in Musca domestica. AB - Musca domestica, the common house fly, has a simple yet powerful and accessible vision system. Cajal indicated in 1885 the vision system is the same as in the human retina [1]. We have modeled the intracellular connections of the vision system and have applied high resolution images to the model. Even though the animal does not have what we would consider a high resolution imaging system, it is still capable of high resolution and we are using the same processing principles to extract high resolution information from the visual input. In this paper we will discuss how we are using a purely local process from a single cartridge within the vision system to encode edge information: magnitude, orientation, displacement, and tilt. We also investigate a cooperative approach to link cartridges that have common numerical features in their field of view to share their information for potential edge connection and feature extraction. PMID- 15133992 TI - Light intensity appears to be more important than an endogenous seasonal clock for regulating structural rhythms in the lateral eye of the horseshoe crab. AB - The lateral compound eye of the American horseshoe crab, Limulus polyphemus, is an established model system for studying visual and circadian processes. Previous studies have shown there is an endogenous rhythm in retinal structure and this rhythm is enhanced by light. The purpose of this study was to determine whether changes in natural lighting influence these rhythms independent of season. To test this concept, horseshoe crabs were maintained in an aquarium tank and exposed to natural lighting only. The angle of the sun and foliage outside the room where the horseshoe crabs were maintained were such that more natural lighting reached the aquarium tank during winter days than summer days despite the fact that sunlight is more intense in the summer. Horseshoe crabs were sacrificed 2 hours, 5 hours, and 8 hours following local sunrise on a winter day (January 11th) and a summer day (June 19th), respectively, and the dimensions of several retinal parameters were quantified. Preliminary data indicate that certain structural rhythms were significantly more pronounced on the winter day when natural lighting was more intense. For example, 8 hours following sunrise, apertures were 4.2 times as long and 3.2 times more narrow on the winter day than on the summer day. These data suggest that the intensity of natural lighting is more important for regulating structural rhythms in the lateral eye than an endogenous seasonal clock. PMID- 15133993 TI - 3-D physical models of mitosis (with asters) and cytokinesis. AB - First, we define new concepts of Life Objects, Informative Objects and Virtual Objects, Discrete Chromosome Rings (DCR); we introduce a mathematical concept of meridian plane (MP) in a three dimensional (3-D) cylindrical coordinate system (CCS). Based on these concepts, classic mechanics, classic electromagnetism and published biological data, we develop our 3-D physical models of natural and normal mitosis (with asters) and cytokinesis, for animal cells in M phase. We propose following hypotheses: Chromosomes Exclusion: No normally and naturally replicated chromosomes can occupy the same nucleus without growing sizes of the nucleus and the cell. Spontaneous and strong electromagnetic fields (EMF) forces among chromosomes, centrosomes and microtubules split the nucleus and separate the two sets of sister chromatids when they are strong enough. Nuclei Exclusion: No normally and naturally doubled nuclei can occupy the same cell if the doubled size of nuclei is not far smaller than size of the cell. The spontaneous and strong EMF forces in protoplasm (or cortex), separate two sets of chromosomes, spindles and poles, drive contractile proteins to the equator in cell cortex, and continue to guide and to transport free charged objects until complete the cytokinesis. Centrioles Exclusion: No naturally and normally doubled centrioles can occupy the same centrosome. The spontaneous and strong repulsive EMF forces are the primary cause for the exclusions. The principles of our models are also applied to mitosis and cytokinesis for lower plant cells, to that of multiple nuclei or mutant chromosomes, and to meiosis, for both animal cells and lower plant cells. PMID- 15133994 TI - The use of digital technology to asses the severity of the Experimental Allergic Encephalomyelitis (EAE) spinal cord lesion. AB - Experimental Allergic Encephalomyelitis (EAE) is an autoimmune demyelinating disease of the central nervous system that is widely accepted as an animal model for the human multiple sclerosis (MS). Since the EAE is also characterized by inflammation in the form of perivascular cuffing, we hypothesized that measuring the number of the inflammatory cells might correlate with the clinical signs and symptoms of the disease and could bring more understanding to the EAE different degrees of the inflammatory process. The model chosen to test this hypothesis was the EAE induced in Lewis rat. The 16 animals were administered Myelin Basic Protein (MBA) plus complete Freund's adjuvant and examined daily in a blind fashion for the signs of MS. Twenty-nine days later, animals were sacrificed and tissues from the spinal cord and were collected for Hematoxylene and Eosin histological studies using the image-Pro-Plus digital morphomtry technique. We concluded that EAE inflammation can be classified into mild, moderate and severs lesion based on the lymphocytes number which correlate with the clinical presentation of the diseased animal. PMID- 15133995 TI - A reduced ambiguity lexical system. AB - Natural human languages have proven to be sub-optimal in artificial intelligence applications because of their tendency to inexact representation of meaning. The author has devised a technique for converting human language to and from a compact byte-coded intermediate representation, which is processed more easily by computer systems. A specialized lexical engine based on IEEE Standard 1275-1994 was created to embed redundant information invisibly within the byte-coded text stream, to enable use of a variety of alphabets, grammars, and pronunciation rules (including slang and regional dialects). Very large vocabularies in a variety of human languages are supported. These lexical tools are designed to facilitate speech recognition and speech synthesis subsystems, universal translators and machine intelligence systems. PMID- 15133996 TI - Static and quasi-static calibration of a bio-MEMS device. AB - A bio-MEMS device that has been designed to stimulate cells by oscillatory actuation in the vertical direction has been calibrated. The displacement of this device was determined experimentally by a laser interferometer when actuated by a static voltage, and by an atomic force microsope when actuated quasi-statically at 0.1 Hz. Both experimental calibrations were compared to a simple model. PMID- 15133997 TI - Eye movement detector calibration device. AB - Presented is a device developed for specifically calibrating and validating the operation of Eye Movement Detectors or Monitors. The Calibrator centers on two one inch diameter HPDE spheres representing the eyes. A Laser Module is embedded in the rear of each sphere emitting a beam against a target divided in equal measurement intervals mounted as part of the device. The device moves the "eyes" about its center axes enabling the user to validate any vertical, horizontal, or X-Y combination eye position in a plus or minus fifteen degree range. Although hand controlled, the Calibrator can be motorized with stepper motors or other desired drivers. Anatomically correct sized pupils are imbedded in the front of each "eye," thereby acting as the target for whichever system is under test by the very portable Calibrator. Currently, a simple battery controlled circuit controls the laser modules and other electric requirements with accommodation for additional circuit components if required in the future. Specifically designed for validating the operation of an IR Reflective Differencing Saccadic Eye Movement Measurement System, the Calibrator can also be used with little or no alteration for validation of camera systems and other types of devices. PMID- 15133998 TI - Mechano-electrical transduction in the turtle utricle. AB - Two classes of mechano-sensory hair cell are present in the vestibular system of mammals, birds and reptiles. Type I hair cells are bottle-shaped and make synaptic contact with afferent calyx terminals. Type II hair cells are cylindrical and contact small bouton afferent terminals. Voltage-dependent basolateral currents have been found to differ between the two cell types and these properties are believed to contribute to the shaping of primary afferent responses. Type I hair cells have a low input resistance, which may be related to the low gain of calyx afferents. In the turtle utricle, type I vestibular hair cells are found only in a narrow band of the sensory epithelium called the striola, whereas type II hair cells are found in both striolar and extrastriolar regions. We have made whole cell patch clamp recordings from type I hair cells, type II hair cells and calyx fibers in order to better understand the processing of vestibular signals. Here we describe responses of hair cells to hair bundle displacement with a stiff glass probe. Mechano-electrical transduction (MET) currents were largest in type I hair cells, where the mean peak amplitude was approximately 500 pA. MET currents in all hair cells showed rapid and slow adaptation. PMID- 15134000 TI - Design of instrumentation and data-acquisition system for complex admittance measurement. AB - Instantaneous left ventricular volume measurements have been made for many years using a tetrapolar conductance catheter. The main objective is to determine the efficiency of the beating heart, using a tetrapolar catheter inserted in the left ventricle of transgenic mice. The effect of the parallel myocardium contribution must be removed from the total measurement. A dual-frequency technique involving 1 kHz and 100 kHz was chosen because it has been established that the imaginary part (the capacitive reactance) of the complex admittance of the cardiac muscle is much smaller in the lower frequency than at the higher frequency. The design involves generation of an accurate frequency source for both the frequencies careful selection of operational amplifiers for the current conversion stage so that the current is not too large to kill the mouse and that it is capable of performing at high frequencies. The band pass filter stage involved careful design with minimal overlap of the pass bands of both the channels. The overall circuit was designed so that there is minimal shift in the phase due to the circuit elements alone. Work also involved design of GPIB--based data acquisition system using LabVIEW and a digital oscilloscope for effective data acquisition even at high frequencies, which are normally limited by the sampling frequency. This data acquisition system is currently being used in laboratory studies in vivo. PMID- 15133999 TI - University of Wyoming, College of Engineering, Undergraduate Senior Design Project: the talking hand. AB - A glove was built using flex sensors to produce a voltage according to the amount of bend each finger produces when signing a letter of the alphabet. The glove detects and outputs in text the letter of the alphabet being signed as the wearer signs the different letters. The amount of bend causes a change in resistance, which in turn produces a specific voltage in accordance to the letter being signed. That voltage is then fed into a data acquisition card that runs into a personal computer. Through intensive programming and training of a special algorithm called a neural network; the input voltage to the data acquisition card will result in that letter being displayed in font on the monitor of the computer. The computer is then programmed to take the text that is displayed on the monitor and run it out of the PC into a store bought chipset that will convert the text to speech. Therefore, a person will be able to put on this glove, sign all twenty-six letters of the alphabet, see the letter they are currently signing output on the monitor, and hear it spoken in a pre-recorded voice. PMID- 15134001 TI - A sleep apnea syndrome detection system. AB - Sleep apnea syndrome (SAS) is a sleep disorder that causes a patient to intermittently stop breathing for longer than 10 seconds and more than 30 times in seven hours. We have developed a new system for non-invasive detection and analysis of SAS. The system consists of a piezoelectric sensor, a microcontroller, a compact flash memory and a laptop computer. The piezoelectric sensor, whose electrical polarization voltage is produced by mechanical strain, is attached to the patient's chest and detects body movements. The low frequency components, mainly generated by respiration, are stored in the compact flash memory. After recording, the data are downloaded to a laptop computer, which then detects whether the patient is in SAS from the amplitude and frequency components of the timed thoracic body movement changes. The system can be used at home and be self-applied by the patients. PMID- 15134002 TI - Monitoring eye movement with a computer based Electro-oculogram (EOG). AB - This project investigated ways to improve retinal laser surgery techniques. Currently, the surgery is manual and not very accurate because the eye has a tendency to move involuntarily and the laser can cause permanent damage to the patient if the eye moves out of range. Therefore, an Electro-oculogram (EOG) system was designed and tested capable of recording eye movement in both horizontal and vertical directions using the voltages produced by the eye for the purpose of feedback to the surgeon. The EOG system consisted of various filters and amplifiers to amplify the small signals created by the eye, which were obtained using surface electrodes placed around the eyes. An LED board was also designed for the subject to follow various LED lights with his/her eyes in all directions. A program was written with LabView software that was used to control the LED board as well as record and analyze the EOG signals. MatLab was used in correlation with LabView to perform additional filtering and to find the mean, root square mean, and standard deviation of the signal. This data was used to determine if a relationship exists between the direction of eye movement and the voltage produced by the eye. The EOG system and LED board operated as expected and the LabView program was capable of recording and analyzing the signals. The information gathered from this project could make retinal laser surgery more precise and safer by alleviating the potential for error and therefore unintentional damage to the patient. PMID- 15134003 TI - Noninvasive treatment of inflammation using electromagnetic fields: current and emerging therapeutic potential. AB - Magnets, electric current and time varying magnetic fields always have played a role in human medicine. Natural magnetic stones were used in ancient cultures to induce a therapeutic effect and modern clinical practice would be far less effective without nuclear magnetic resonance imaging, cardiac pacemakers, and bone growth stimulators. This paper presents a summary of natural and artificial electromagnetic field (EMF) characteristics that are currently in use or under investigation for other therapeutic applications. This background understanding provides a basis for discussion on the success and possible risks of emerging and novel EMF therapies. Although interest in energy medicine has existed for centuries in some parts of the world, in recent years this is an area of heightened interest for western healthcare practitioners. This awareness has been triggered by the growing body of knowledge on how EMFs interact with cellular systems. EMF therapy for the treatment of pain, cancer, epilepsy, and inflammatory diseases like psoriasis, tendinitis and rheumatoid arthritis is currently being explored. The long-term success of this new area of medicine is still unknown. On the one hand, it remains to be seen whether positive human outcomes with EMF therapy could be explained by enhancement of the placebo effect. Optimistically, EMF therapy has the potential to revolutionize medicine, which is currently dominated by pharmaceutical and surgical interventions. In this case, new therapeutic tools may be developed for future clinicians to provide noninvasive treatments with low risk of side effects and no problem with drug interactions. PMID- 15134004 TI - Sending data to a central repository. AB - The Veterinary Medical DataBases (VMDB) has been collecting data from participating institutions for 40 years. We have recently started a program to collect clinical case data from not only veterinary schools but from veterinary practitioners as well. In so doing, we are using commonly accepted means of data representation and transmission. Thus, we use the Standard Nomenclature of Medicine (SNOMED) as a standard vocabulary language and Health Level 7 (HL 7) as a standard transmission protocol. We report on the planning and implementation of these standards and the problems encountered in their implementation. PMID- 15134005 TI - A welfare facility resident care support system. AB - We have developed a new care support system to monitor the overall health of welfare facility residents who need constant care. The system is designed with wireless sensors, wireless repeaters and a host computer. The wireless sensor, which consists of a piezoelectric sensor, a 2 axis accelerometer, a microcontroller and a low power 303 MHz ASH transceiver. It records respiration, activity and indicators of posture and behavior for 24 hours. These data are transmitted to the wireless repeater by the transceiver. The wireless repeaters, which are installed throughout the welfare facility, send data, including the repeater's ID, to the host computer. The ID is used to detect the resident's location in the welfare facility. The host computer stores the data, which can be used to analyze the resident's overall health condition. When the resident is in an emergency situation, such as falling or in an inactive state for more that the allotted time, the host computer automatically alerts the situation to the care staff by an alarm sound and also by mobile phone. PMID- 15134006 TI - The University of Connecticut Biomedical Engineering Mentoring Program for high school students. AB - For the past four years, the Biomedical Engineering Program at the University of Connecticut has offered a summer mentoring program for high school students interested in biomedical engineering. To offer this program, we have partnered with the UConn Mentor Connection Program, the School of Engineering 2000 Program and the College of Liberal Arts and Sciences Summer Laboratory Apprentice Program. We typically have approximately 20-25 high school students learning about biomedical engineering each summer. The mentoring aspect of the program exists at many different levels, with the graduate students mentoring the undergraduate students, and these students mentoring the high school students. The program starts with a three-hour lecture on biomedical engineering to properly orient the students. An in-depth paper on an area in biomedical engineering is a required component, as well as a PowerPoint presentation on their research. All of the students build a device to record an EKG on a computer using LabView, including signal processing to remove noise. The students learn some rudimentary concepts on electrocardiography and the physiology and anatomy of the heart. The students also learn basic electronics and breadboarding circuits, PSpice, the building of a printed circuit board, PIC microcontroller, the operation of Multimeters (including the oscilloscope), soldering, assembly of the EKG device and writing LabView code to run their device on a PC. The students keep their EKG device, LabView program and a fully illustrated booklet on EKG to bring home with them, and hopefully bring back to their high school to share their experiences with other students and teachers. The students also work on several other projects during this summer experience as well as visit Hartford Hospital to learn about Clinical Engineering. PMID- 15134007 TI - Supporting undergraduate biomedical entrepreneurship. AB - As biomedical innovations become more sophisticated and expensive to bring to market, an approach is needed to ensure the survival of the best ideas. The tactic used by Iowa State University to provide entrepreneurship opportunities for undergraduate students in biomedical areas is a model that has proven to be both distinctive and effective. Iowa State supports and fosters undergraduate student entrepreneurship efforts through the Pappajohn Center for Entrepreneurship. This unique partnership encourages ISU faculty, researchers, and students to become involved in the world of entrepreneurship, while allowing Iowa's business communities to gain access to a wide array of available resources, skills, and information from Iowa State University. PMID- 15134008 TI - Nursing law and rules in Ohio: an overview. PMID- 15134009 TI - What RN's should know about organizing. PMID- 15134010 TI - We have new residents that sometimes forget to sign off the orders of the medical students. PMID- 15134011 TI - Safe staffing and mandatory overtime: issues of concern in today's workplace. PMID- 15134012 TI - My hospital serves the minority populations in the surrounding area. Many of these patients are immigrants of different ethnic backgrounds. PMID- 15134013 TI - What every RN should know about a grievance--Part Three. Grievance presentation and negotiation. PMID- 15134015 TI - Does the whistleblowing law protect me? PMID- 15134014 TI - The ethical decision making process: a demonstration. AB - Two issues identified from the scenario have been explored using the Ethical Decision Making Tree. Now look at some of the other issues identified earlier in this article work through the decision-making process using the Ethical Decision Making Tree. In looking at the use of experimental or futile care, what information would you need to gather and analyze? What personal values and moral positions need to be considered? Based on the previous two steps, what are the options? What is your decision and course of action? How are you going to evaluate the action taken to see if it worked or if other actions need to be taken? Remember, the solution is not what is best for you but what is best for the patient at this point of time. PMID- 15134016 TI - What is a peer review organization anyway? What nurses need to know. PMID- 15134017 TI - HIPPA general overview. Part I. PMID- 15134018 TI - Obtaining informed consent for psychotropic drugs to the nurses. PMID- 15134019 TI - Preventing violence in the healthcare workplace. PMID- 15134020 TI - Employers are required to provide unions information. PMID- 15134021 TI - I have been fired from my job. My employer says that I will be reported to the Board of Nursing and may face criminal charges. PMID- 15134022 TI - Hospital Issued Notice of Non-payment (HINN): what is it? (when Medicare doesn't pay). PMID- 15134023 TI - I am a registered nurse in desperate need of some straight forward answers relating to the legality of an LPN being "in charge" of an RN. PMID- 15134026 TI - Ohio Nurses Association union security notice. PMID- 15134024 TI - HIPAA--a matter of privacy. PMID- 15134029 TI - In the state of Ohio, is a nurse legally responsible to stop at the scene of an accident and provide medical help until the proper people arrive? PMID- 15134030 TI - [Nutritional support of patients treated with continuous hemofiltration methods in intensive care units]. AB - The use of continuous renal replacement therapy (CRRT) can directly or indirectly influence patient's metabolism and nutritional status. The loss of nutrients into the filtrate is significant. It concerns aminoacids, glucose, water-soluble vitamins, some minerals and trace elements. The loss of aminoacids is about 1.2 to 7.0 g per day. The glucose balance during CRRT depends on the concentration of glucose in substitution solutions. When concentration between 80 to 180 mg/dl is used, the glucose balance in most of patients is kept. The loss of lipids and lipid-soluble vitamins is neglectable. From the micronutrients, significant becomes the loss of magnesium, calcium, phosphorus and that of water-soluble vitamins (mainly B1, B6, C and folic acid). Contrary to it, some components can cross from the substitution solution to the blood (e.g. lactate). Indirect effect of CRRT on the nutrition and metabolism represents the release of cytokins and inflammatory mediators resulting from the first contact of the blood with the filter membrane. The decrease of glutamine level in the beginning of therapy is probably a consequence of this inflammatory reaction. Nutritional plan of most of patients treated with CRRT is therefore necessary to adjust. PMID- 15134031 TI - [The large intestine--literature review of recent years]. AB - Diagnose of the idiopathic inflammatory bowel disease depends on the set of clinical, radiological, endoscopic and histological criteria. Proctitis, proctosigmoiditis, and the left-sided colitis represent 60 to 80 percent of newly diagnosed cases of colitis. For the medical management of the inflammatory bowel diseases aminosalicylates, corticosteroids, topical corticosteroids, immunosuppressants are used. The complication of highest risk represents the toxic magacolon or periannal fistulas in patients with Crohn's disease. Pre neoplastic epithelial changes (dysplasia) are diagnosed by pathologists according sets of cytological and architectonic markers. Indeterminate diarrhea related to the antibiotics administration is comparatively common. Roman criteria II classify functional intestinal disorders, represented by a set of isolated symptoms or various combinations of symptoms without identifiable organic, biochemical, sonographic or endoscopic abnormalities of the intestinal or biliary system, lasting minimally 12 weeks (not necessarily continuously) during the preceding year, as a persisting or recurrent distress. About 20 percent of patients with diverticular disease of the colon have some clinical manifestations. PMID- 15134032 TI - [Infectious diseases in 2003]. AB - A brief survey of the present incidence of infectious diseases (by WHO) and new possibilities of their team study. Global significance of morbidity and mortality of some infections is stressed, namely those of malaria, tuberculosis, HIV infection, infantile diarrhoea, respiratory diseases. Alarming trends in age distribution of some diseases (87% of HIV infection affects young population between 15 to 19 years) or the significance of HIV infection in combination with tuberculosis in previous 10 years in Africa are pointed out. Serious appears the information on the pharmaco-resistance of mycobacteria as well as the growing resistance of Plasmodium falciparum to chlorochin, namely in the regions of the Southeast Asia. PMID- 15134033 TI - [Genetic risk factors in autoimmune diabetes mellitus, their significance and function]. AB - Autoimmune diabetes mellitus is characterized by selective destruction of beta pancreatic cells and by cellular infiltration with T- (particularly Th1) and B lymphocytes. The marker of autoimmunity is the presence of autoantibodies (ICA, IAA, GADab, IA2ab). Etiology of the autoimmune process is still unknown. It is suggested that the pathogenesis is activated by genetic and environmental factors. Individual predisposition can influence also the onset and progression of the disease. The most important genetic risk factors of autoimmune diabetes mellitus are the HLA class II alleles (DQB1*0302, 0201; DRB1*0301, 0401; DQA1*0301, 0501) and the risk alleles of INS-VNTR of the promoter region. Recent studies have shown various genetic risk factors for the autoimmune diabetes mellitus. Individual predispositions belong to the genetic polymorphisms in cytokine genes (IL-10, IL-12, IL-18) and the microsatellite polymorphism of MHC class I chain-related gene A (MIC-A). PMID- 15134034 TI - [Loss of intelligence in aphasic disorders]. AB - BACKGROUND: Certain level of aphasia is considered to be an integral component of dementia. More controversial is the question whether every--e.g. also strictly focal--aphasia must be accompanied with dementia. METHODS AND RESULTS: 41 persons hospitalized after the cerebral stroke (17 men, 24 women, average age 64.6, 47 to 89 years old) were tested to correlate the level of aphasia with mental and intelligence scores. 35 patients had an ischaemic stroke, 6 had a hemorrhagic one; according to CT/MRI there were 37 patients with left sided impairment, one patient with a right sided one and 3 patients with bilateral lesions. As an aphasiologic test the Western Aphasia Battery (WAB) was used, to determine the general mental performance as an equivalent of IQ test the non-verbal Raven's and verbal Euro-ADAS test were used. Correlation analysis using non-parametric Spearman's coefficient showed a significant relation between results in phatic functions and the score in intelligence tests. Phatic performance had in all elementary tests closer relation to verbal IQ in Euro-ADAS than with non-verbal IQ in Raven's test. The highest correlation was found in the relation between "apprehension vs. Euro-ADAS" (-0.842) and "apprehension vs. Raven" (0.782), while the lowest correlation was found for "repetition vs. Euro-ADAS" (-0.769), and "repetition vs. Raven" (0.646). The correlation was in all cases significant (p < 0.00001). Similar narrow relation occurred between the clinical forms of aphasia and intelligence tests. Sensory aphasia was associated most closely with both relations, with Euro-ADAS (-0.800), and Raven's test (0.800), while global aphasia had the lowest correlation with Euro-ADAS (-0.451) and with Raven's test (0.517). CONCLUSIONS: Speech is not only a product of thinking but also represents a source and structural tool for thinking. PMID- 15134035 TI - [Spindle cell and cuboidal renal cell carcinoma (loopoma). 10 case reports]. AB - BACKGROUND: Current classification systems of neoplasms arising from renal parenchyma distinguish 5 categories of renal cell carcinoma (RCC), i.e. conventional RCC, papillary RCC, chromophobe RCC, collecting duct/medullary RCC and unclassified RCC. We present 13 cases of unusual and unclassified spindle and cuboidal renal cell carcinomas. METHODS AND RESULTS: The studied group consisted of 13 patients (7 men and 6 women). They ranged in age from 22 to 65 years (mean 57.3). Generally, the tumours were well circumscribed and confined to the kidney, whitish to grey on section with a diameter 4.5-13 cm (mean 8.6 cm). One patient was investigated for loin pain and nocturia. Three patients had staghorn nephrolithiasis and vague sonographic findings in renal parenchyma. In one patient the renal tumour was found when examined on follow-up examination for prostatic adenocarcinoma. None of our patients was known to have elevated levels of parathyroid hormone due to hyperplasia, adenoma or carcinoma of the parathyroid gland. Clinical follow-up of the patients ranged from 9 months to 8 years (mean 2.3 years). Microscopically, the tumours were composed of two main populations of cells: flattened, spindle cells with sparse cytoplasm and small cuboidal cells with clear to light eosinophilic cytoplasm. Eight patients are currently well without signs of recurrence or metastasis, one had metastasis in the regional lymph node at the time of nephrectomy, one died of unrelated cause, and three were lost to follow-up. CONCLUSIONS: We present 13 cases of unclassified RCC. Our cases were histologically, immunohistochemically and ultrastructurally similar to the hitherto reported case reports of this variant of RCC. It is obvious, that that variant of RCC should be recognised as a new subtype of RCC. PMID- 15134036 TI - [Clinical importance of determination of tumor markers during follow-up in breast carcinoma]. AB - BACKGROUND: Paper deals with detection of the early disease progression in breast cancer patients during follow up using tumor markers. METHODS AND RESULTS: The basic group of 1184 patients with breast carcinoma in follow up after primary therapy were examined from 1996 to 2002. Sera were tested using commercial kits CA 15-3 (MEIA, Abbot), CEA (IRMA, Immunotech), TPA (IRMA, Byk Sangtec), TPS (IRMA, Beki). Results were compared with the retrospectively confirmed clinical status of individual patients. The authors calculated optimal cut offs and sensitivities and their combinations for particular tumor markers at 95% level of specificity. Best sensitivities for the detection of distant metastases into bone, liver, lung and brain was achieved by CA 15-3 (53-68%). As an optimal combination of tumor markers seems to be the tricombination CA 15-3, CEA and TPA. All the tumor markers have insufficient sensitivity for the metastatic process into the lymphnodes. CONCLUSIONS: As optimal combination of tumor markers during the follow up seem to be tricombination CA 15-3, CEA and TPA, but also the clinical relevance and cost effectiveness of these assessments have to be considered. For the tumor disease follow up only CA 15-3 has sufficient sensitivities (at 95% specificities) for the early diagnosis of the metastatic process. PMID- 15134037 TI - [Antibiotic resistance in invasive pneumococci and their serotypes in the Czech Republic]. AB - BACKGROUND: Among strains of S. pneumoniae (pneumococci), isolated from blood or cerebrospinal fluid of 483 patients between January 2001 and October 2003 as many as 5.2% strains were penicillin-non-susceptible. METHODS AND RESULTS: Incidence of penicillin-non-susceptible S. pneumoniae (PNSP) strains was highest in children of the youngest age group (12.7%). Cefotaxim-non-susceptibility was detected in 2.7% of strains. Pneumococci tested were resistant to co-trimoxazol (9.9%), tetracycline (8.9%), chloramphenicol (4.6%), erythromycin (2.3%), clindamycin (1.4%), levofloxacin (0.6%) and rifampicin (0.2%); none of the strains was resistant to linezolid. Comparison of the results from 1996-1999 shows a permanent occurrence of invasive PNSP strains ranging about 5%. The PNSP strains were of 7 serotypes, with serotype 9V being the most frequent. The Czech collection of invasive pneumococcal strains is five times smaller than that of any other European country with comparable populations over the same period of time. CONCLUSIONS: Low incidence of invasive S. pneumoniae strains seems to be related to the underestimated significance of blood culturing. Valid data necessary for the safety of antibiotic therapy in terms of efficacy and prevention of resistance development cannot be obtained from some localities of the Czech Republic. PMID- 15134038 TI - [Clinical study of Hodgkin's disease after 25 years]. AB - BACKGROUND: On the 25th anniversary of the unified protocol of 1978, the authors report the results of the first clinical study in the Czech Republic performed in patients with Hodgkin's lymphoma. 290 patients were treated with both chemotherapy (Cyklophosphamide, Natulan, Prednison, Vinblastin) and radiotherapy according to this widely used protocol. The dosage of cytotoxic therapy was reduced based on age, in patients over 50 years of age. METHODS AND RESULTS: In the year 2003, the survival rate was 77.3% (194/290 patients) and of these, 85% (165/194 patients) have lived more than 10 years with complete remission. Of the 194 surviving patients, 24 were successfully treated for relapses. Of the 66 (22.8%) patients that died, the cause of death in 33 patients was Hodgkin's lymphoma. 33 women of the studied group gave birth to 43 children. In one case, phocomelia was diagnosed post-partum. In this instance, no causal relationship to the cytotoxic treatment (which had been given three years before the birth) could be established. The second child of the same mother was born healthy. In one case, a possible genetic relationship was noted--both the father and the daughter suffered from Hodgkin lymphoma and the granddaughter from non-Hodgkin lymphoma. CONCLUSIONS: The success of this treatment depends on complex diagnostic procedures, and on an experienced team of physicians. PMID- 15134039 TI - [Evaluation of three dosage regimens of amikacin using pharmacokinetic models in patients with cystic fibrosis]. AB - BACKGROUND: Once-daily administration of aminoglykosides is routinely used, but comparative efficacy data for patients with cystic fibrosis are not available. METHODS AND RESULTS: The aim of the this study was to compare the predicted pharmacodynamic (PD) activity of amikacin at 28 mg/kg/den administered every 24 hod.(q24h), q12h, and q8h. Pharmacokinetic (PK) data were derived from analysis of the amikacin serum concentration from 42 CF children patients. Individual pharmacokinetics values were used to construct serum concentration--versus time curves and to determine various indices (c peak/MIC ratio and time during the concentration was less than the MIC--T < MIC) for all three dose regimens described above. MIC (minimal inhibitory concentration) for Pseudomonas aeruginosa was 4 mg/l. Significantly lower c peak/MIC but shorter T < MIC were noted when regimens of q8h versus q12h (p < 0.001), q8h vs. q24h (p < 0.001) and q12h vs. q24h (p < 0.001) were compared. This analysis suggests that the potential advantage of achieving a greater c peak/MIC with once-daily aminoglycoside administration may be neutralized by the significantly greater T < MIC in CF patients compared with that achieved with multiple-daily-dosing regimens. CONCLUSIONS: Routine use of once daily amikacin administration could not be recommended until the clinical data confirming efficiency of this dose modality are available. PMID- 15134040 TI - [Molecular diagnosis of malignant B-cell lymphoma: detection of clonality and a brief review of problems with our results]. AB - BACKGROUND: Diagnostic approach to non-Hodgkin's lymphomas requires a combination of laboratory methods. Methods used in morphology constitute the basis of the diagnostics; in many instances it is necessary to combine them with methods of molecular genetics. The latter method plays a key role in the detection of B cell clonality using identification of the rearrangement of IGH and/or IGK genes and in detection of the chromosomal translocations specific for some lymphomas. METHODS AND RESULTS: Using PCR we investigated 113 patients with malignant B cell lymphomas of different types (follicular--FL, mantle cell--MCL, small cell- CLL/SLL, diffuse large cell--DLBCL). We established the IGH gene clonal rearrangement in 85% of the cases (96/113), and the clonal rearrangement of the IGK gene in 58.3% patients (42/72). Combination of both approaches (IGH and IGK) revealed a positive result in 90.3% (102/113). The highest yield was rendered in patients with CLL/SLL and with MCL (100%), and it was 86 and 87% in cases with FL and DLBCL. CONCLUSIONS: The detection of clonality in lymphomas helps to distinguish a malignant disease from polyclonal hyperplastic and lymphoproliferative disorders of B cells. The recognition of clonal rearrangements of the IGH and IGK genes serves for a long term monitoring of the disease activity in cases in which there are no other molecular markers available. The demonstration of lymphoma characteristic translocations is relatively specific and useful but at present its usefulness is reduced in cases with variable breakpoint regions. PMID- 15134041 TI - [Angiomyolipoma--its role in prenatal diagnosis of tuberous sclerosis]. AB - Tuberous sclerosis (TSC) is a frequent hereditary autosomal-dominant disease characterised by hamartomas developing in many organs. The disorder is caused by mutations affecting either of the tumor-suppressor genes, TSC1 and TSC2. Tumorogenesis is triggered by the loss of second functional gene copy, mostly accompanied by loss of heterozygosity (LOH) of flanking polymorphic markers. Search for causing mutations is very laborious, time consuming and low effective. Prenatal diagnosis is often hampered by lack of detection of causing mutation. Detection of LOH in hamartomatous tissue suggests which gene is involved in particular case of disease and specifies which of homologous chromosomes carries germinal mutation. Examination of LOH is useful for prenatal diagnostics especially when time is lacking due to patient's pregnancy or in case of mutation screening failure. PMID- 15134042 TI - [The Budd-Chiari syndrome in a patient with primary thrombocythemia treated with interferon alfa and transjugular portosystemic shunt]. AB - The case report of a young female patient with personal history of primary thrombocythaemia, treated with interferon alpha, admitted to our medical department for severe abdominal pain, hepatomegaly, ascites and alteration of hepatic function is presented. Magnetic resonance imaging showed the picture typical for Budd-Chiari syndrome caused by external obstruction of the intrahepatal portion of inferior vena cava. The cause of the syndrome remains uncertain, possibility of the haematogenic infiltration of the liver or venal thrombosis within primary or secondary (interferon-induced) antiphospholipid syndrome is discussed. Liver biopsy could elucidate the exact cause, but it was not performed for technical problems. PMID- 15134043 TI - [University qualifying work (dissertations), their current registration, electronic access and related technical, administrative and legal questions]. AB - Many countries (USA, Australia, Canada, Latin America, Spain, France, SRN) have national or international systems (e.g. Networked Digital Library of Theses and Dissertations--NDLTD), which openly (NDLTD) or commercially (University Microfilm International--UMI) present up to 50% of master dissertations and doctoral theses in a full text form. Such systems are in agreement with legal regulations of the given country and the author is the only holder of the work--of the dissertation. Authors are at the same time informed that dissertation represents a university document, which presents results of the given scientific community and it should be therefore distributed free of charge. Presentation is also the matter of honour of the author and his university. In the contemporary Czech Auctorial Law (No. 121/2000 Sb.) dissertation is considered as not fully defined "Educational work" and from the practical point the duration of necessary storage is not solved. For the contemporary worldwide conversion to electronically presented dissertations, the necessary technical and library standards has been prepared with the first positive experience. The supplement of the Auctorial and University Law appears to be necessary. It is inevitable to be prepared for that situation before the Czech Republic will join EU. PMID- 15134044 TI - [Quality control. Message to laboratories--do not fear accreditation, message to patients--demand it]. AB - Though the strain for the highest possible quality is the implied part of the health care, certain formalization is possible. It means to name its components, to give the aims and methods how to achieve them, to follow and evaluate their fulfillment. In other words: Through the accreditation we can achieve our effort to be visible and sustainable. PMID- 15134045 TI - [Smoking and reproduction. Part 1. Smoking and sexual health]. AB - The first of the three-part series deals with effects of smoking on sexual and reproductive health. The second part will be published the next month, and it will consider smoking and pregnancy. Part three will concern effects of smoking on some other aspects of reproductive health. PMID- 15134047 TI - D. Neil Granger. 77th president of the APS. PMID- 15134046 TI - [Whatever is free is worthless. On the article "Health and money". ]. PMID- 15134049 TI - Contract enforcement. PMID- 15134048 TI - Asthma. PMID- 15134051 TI - I have been fired from my job ... What are my rights? Why wasn't I given due process? PMID- 15134052 TI - Safer nurses; safer care: ban on mandatory overtime proposed. PMID- 15134053 TI - A nursing malpractice primer. PMID- 15134054 TI - Night shift nurse was videotaped as she conducted a client interview in segregation. PMID- 15134055 TI - ONA minority nurses reflect on the current nursing shortage. PMID- 15134056 TI - HIPPA general overview. Part II. PMID- 15134059 TI - Interpreting lab values affected by kidney function. AB - Renal function and fluid and electrolyte balance go hand in hand. Many health conditions affect renal function so it is important that nurses understand this relationship. Failure to do so might result in missing important signs and symptoms that can provide the key to the diagnosis and treatment of many patients. PMID- 15134061 TI - Smallpox: what every nurse should know. PMID- 15134060 TI - Where can I find the nursing practice standards for assisted living facilities? PMID- 15134062 TI - The Family Medical Leave Act--know your rights! PMID- 15134064 TI - Affordable prescription drugs--state and federal action. PMID- 15134063 TI - The role of the LPN in assessment. PMID- 15134067 TI - Molecular mechanisms of receptor activation: real-time analysis by fluorescence resonance energy transfer. PMID- 15134068 TI - Brain meets pancreas: netrin, an axon guidance molecule, controls epithelial cell migration. AB - A recent publication shows that two integrins, alpha6beta4 and alpha3beta1, interact with netrin-1, a protein best known for its role in axon guidance. Consistent with a role in pancreatic development in vivo, netrin is expressed together with these integrins in fetal pancreatic epithelium. The paper presents strong evidence that pancreatic epithelial cells adhere, spread and migrate on netrin-coated substrata through alpha6beta4. Data also indicate that alpha3beta1 mediates HGF-stimulated cell migration on netrin-coated substrata. These results indicate that netrins might have much broader functions than previously suspected. PMID- 15134069 TI - "Anterior deprogramming device fabrication using a thermoplastic material" (Land, MF, Pregerina, A. J Prosthet Dent 2003;90:608-10). PMID- 15134070 TI - Abusive docs drive out nurses. PMID- 15134071 TI - A happier staff. PMID- 15134072 TI - Clarity on defined contributions. PMID- 15134073 TI - Early use of corrective lenses in Spanish colonies of the Americas including parts of the future United States: reference to Viceroy Luis de Velasco (the son). PMID- 15134074 TI - MicroRNA precursors in motion: exportin-5 mediates their nuclear export. AB - The discovery of microRNSs (miRNAs) has changed the paradigm of gene regulation, leaving us with numerous exciting questions regarding what these molecules do and how they originate. A model for miRNA biogenesis has emerged recently, yet several key factors--including the identity of the miRNA nuclear export receptor- remained unknown. However, recent studies have shown that exportin-5 (Exp5), a Ran-dependent importin-beta-related transport receptor, mediates nuclear export of miRNA precursors (pre-miRNAs). PMID- 15134075 TI - Shaping science policy in the age of genomics. PMID- 15134076 TI - [Otorhinolaryngological aspects of HIV-infection and AIDS]. PMID- 15134077 TI - Structurally speaking. PMID- 15134078 TI - Strength in diversity. PMID- 15134079 TI - [The role of chlamydial and mycoplasmic infection in etiology of ENT diseases]. PMID- 15134080 TI - [Effective pharmacotherapy in endosurgical operations on the maxilla and ethmoidal labyrinth]. PMID- 15134081 TI - Dengue. PMID- 15134082 TI - [Management of ENT diseases complications using ticarcillin clavulanate]. PMID- 15134083 TI - [Safety of a novel preparation -- strepfen -- for symptomatic treatment of pain in the throat]. PMID- 15134084 TI - [Life without limit?]. PMID- 15134085 TI - [New developments as regards coronaviruses]. PMID- 15134086 TI - Regarding "Nomenclature of the veins of the lower limbs: an international interdisciplinary consensus statement. PMID- 15134087 TI - Regarding "Treatment of superficial and perforator venous incompetence without deep venous insufficiency: is routine perforator ligation necessary?". PMID- 15134088 TI - Regarding "Challenges of endovascular tube graft repair of thoracic aortic aneurysm: midterm follow-up and lessons learned". PMID- 15134089 TI - [HIV/HCV coinfection: 40% of patients can successfully be treated]. PMID- 15134090 TI - Regarding "Randomized trial and local biological effect of autologous platelets used as adjuvant therapy for chronic venous leg ulcers". PMID- 15134091 TI - [Lopinavir: also effective using once daily dose in therapy-naive HIV patients]. PMID- 15134092 TI - Regarding "Acute arterial complications associated with total hip and knee arthroplasty". PMID- 15134093 TI - Advanced health telematics and telemedicine. Proceedings of the Magdeburg Expert Summit. December 4, 2002. Magdeburg, Germany. PMID- 15134094 TI - Oral brush biopsy: false positives redux. PMID- 15134095 TI - Dental follicles replantation after enucleation of an odontoma. PMID- 15134097 TI - Mass spectrometric immunoassay. AB - A new, general method of immunoassay is demonstrated. The approach is based on the microscale immunoaffinity capture of target antigens followed by mass specific identification and quantitation using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Immunoaffinity capture of antigens effectively overcomes signal suppression effects typically encountered during traditional matrix-assisted laser desorption/ionization analysis of complex biological mixtures while simultaneously concentrating the analyte into a small volume. Mass spectrometric detection of antigens is unambiguous, as antigen signals are observed at characteristic mass-to-charge values in the mass spectrum, offering a high level of immunity to artifacts due to nonbiospecific retention of mixture components. However, the most important aspect of such mass specific detection is the ability to use a single assay to screen biological systems for the presence of multiple, mass-resolved antigens. Analyte quantitation is possible by using a single antibody to capture both the antigen and an antigen variant which has been chemically modified to have a different mass. With proper calibration, the relative signal intensities of the two species in the mass spectrum can be used to determine the antigen concentration. Sample incubation and processing methods were such that a typical analysis could be performed in less than 1 h while subnanomolar sensitivities were maintained. The technique has been used for the rapid, selective, and quantitative screening of human blood for the presence of myotoxin a, and Mojave toxin form the venoms of the prairie rattlesnakes, Crotalus viridis viridis, and and the Mojave rattlesnake, Crotalus scutulatus scutulatus. PMID- 15134098 TI - Gambling and other risk behaviors on university campuses. AB - To examine the prevalence of problem gambling and its relationship to other risk taking behaviors, the authors surveyed 1,350 undergraduates at the 4 campuses of Connecticut State University (CSU) during fall 2000. On the basis of a modified version of the South Oaks Gambling Screen, a widely used screening instrument, they found that 18% of the men and 4% of the women reported that gambling had led to at least 3 negative life consequences (eg, felt guilty, gambled more than intended), commonly defined as problem gambling. Students identified as problem gamblers, compared with other students, were significantly more likely to be heavy drinkers, report negative consequences of alcohol consumption, and be regular tobacco and marijuana users. Problem gambling was related to binge eating and greater use of weight-control efforts. University athletes were also found to have significantly greater problem gambling rates than nonathletes. The majority of students gambled but experienced few of the negative consequences reported by problem gamblers. PMID- 15134096 TI - [Skin cancer risk after organ transplantation]. PMID- 15134099 TI - Mass-mediated information effects on testicular self-examination among college students. AB - The author assessed the effects of a youth-targeted national cable broadcast that promoted testicular self-examination (TSE). A telephone survey of 524 college men included variables from the theory of reasoned action, as well as measures of self-exam compliance, message exposure, knowledge, attention, and the effects of humor and fear. Exposure to the show, with attitudes and norms controlled for, demonstrated a statistically significant but very weak effect on viewers' behavioral intentions. Although also statistically significant, the effect of the show on awareness, knowledge, and facilitating interpersonal communication was as weak or weaker than its effect on intention. Furthermore, dose measures failed to demonstrate any significant effect among those who saw the show. It is conceivable that the show might be integrated into a broader effort toward TSE education, but the results of this study do not argue that it could be effectively used as a media-only TSE promotion. PMID- 15134100 TI - Partner communication skills and condom use among college couples. AB - The authors examined the effects of (1) communication-skills training, (2) relationship-specific education, or (3) risk information on the condom use of 106 heterosexual college students who were in sexually active relationships. Postintervention assessments of in vivo communication-skills demonstrated that participants in the communication-skills group acquired the skills to use direct requests for condom use and to counter partner refusal statements more effectively than participants in the other conditions (ps < .01). Despite successful skill acquisition, however, students failed to change their subsequent communication or condom-use behavior. This pattern of findings is considered in terms of how the development of sexual partnership in this cohort affects the interplay of motivation and skills for condom use. PMID- 15134101 TI - Athletic status and drinking behavior in college students: the influence of gender and coping styles. AB - College students' alcohol use as well documented, and published studies have indicated that athletes drink more frequently and more often to the stage of intoxication than do nonathletes. Some researchers have cited sociological factors to explain these behaviors, but neither the underlying emotional factors that drive students' alcohol use nor the interaction of gender and athletic status have been examined. The authors' twofold purpose in conducting this study was (1) to examine the influence of the interaction of gender and athletic status on the drinking behaviors of college students, and (2) to examine whether differences in male and female athletes' and nonathletes' coping styles influenced their drinking behaviors. PMID- 15134102 TI - College health services: reflections of a university president, "You are special and you can be even better". PMID- 15134103 TI - Corneal surface changes in Thygeson's superficial punctate keratitis: a clinical and non-contact photomicrographic in vivo study in the human cornea. AB - PURPOSE: To elucidate mechanisms behind the morphology of Thygeson's superficial punctate keratitis (TSPK). METHODS: Sixteen patients were examined with the slit lamp and photographed by non-contact photomicrography. The results were compared with morphology of epithelial keratitis in herpes simplex type 1 (HSV1), varicella zoster (VZV), and adenovirus type 8 (Ad8) infections, all previously studied by the same method, and with published histologic findings in TSPK. RESULTS: In the photographs, the corneal epithelium showed various numbers of abnormal subsurface cells measuring about 10-15 microm in diameter, present individually, in small groups, or aggregated in larger lesions (coarse lesions with the slit lamp). The surface epithelium was well preserved, except in larger lesions, which showed surface debris. The morphology was unlike HSV1 and VZV epithelial keratitis, but strongly resembled epithelial changes occurring in Ad8 infections on day 5, and later, after the onset of symptoms. CONCLUSIONS: TSPK shows a more widespread epithelial involvement than suspected with the slit lamp. Its morphology seems to reflect an action of a noxious agent targeted at the deeper epithelial layers, with the appearance of abnormal cells as a result. These might represent invading inflammatory cells, damaged intraepithelial ones, or both. The coarse lesions visualize areas of major involvement showing discernible signs of cell destruction. The similarity to Ad8 keratitis suggests that the source of the noxious agent might be located outside the cornea. The morphology, in conjunction with clinical features, is compatible with an immunologically mediated injury. The etiology remains unknown. PMID- 15134104 TI - Refractive outcome of single running suture adjustment in penetrating keratoplasty. AB - PURPOSE: To evaluate the effectiveness of single running suture adjustment in reducing postoperative astigmatism and improving visual acuity in patients who have undergone penetrating keratoplasty. METHODS: Eighteen eyes of 18 patients who underwent penetrating keratoplasty at the Department of Ophthalmology, Baskent University between May 1997 and December 2000 and who had an astigmatism over 2 diopters (D) at the fourth postoperative week were included. All eyes underwent suture adjustment at the fourth week and eight eyes with residual astigmatism above 3 D underwent a second adjustment at the eighth week. Final astigmatism and visual acuity findings were evaluated 28 weeks postoperatively. RESULTS: The mean postoperative keratometric and refractive astigmatism were 5.76 +/- 0.88 D (range=4.50-7.76) and 5.22 +/- 0.78 D (range=4.0-7.0) at the fourth week, which were decreased to 2.82 +/- 0.99 D (range=1.25-4.62, p=0.0001) and 2.61 +/- 1.01 D (range= 1.0-4.5, p=0.0001) after suture adjustment, respectively. In eight eyes at the eighth week, a second suture adjustment reduced the average keratometric and refractive astigmatism from 3.86 +/- 0.65 D (range=2.75-4.62) and 3.81 +/- 0.44 D (range=3.25-4.50) to 2.33 +/- 0.91 D (range=0.87-3.50, p=0.012) and 2.06 +/- 0.68 D (range=1.0-2.75, p=0.011), respectively. At the 28th week, the average keratometric and refractive astigmatism levels for the entire cohort, which were 2.39 +/- 1.06 D (range=1.0-3.50) and 2.25 +/- 0.96 D (range=1.0-3.25), respectively, showed a statistically significant decrease when compared with the levels before the first suture adjustment (p=0.0001 for both data). The mean best-corrected visual acuity was 20/25 on the Snellen chart at the 28th week. CONCLUSIONS: Postoperative adjustment of single running suture is a safe and effective way of reducing postkeratoplasty astigmatism. PMID- 15134105 TI - Nitric oxide levels of aqueous humor after photorefractive keratectomy. AB - PURPOSE: To measure the nitric oxide (NO) levels of aqueous humor in rabbits after photorefractive keratectomy (PRK) and to evaluate the alterations of NO levels according to the PRK surgery steps, ablation depth, and time. METHODS: Fifty eyes of 25 New Zealand white rabbits were included in the study. One eye was later randomly excluded from the study in order to equalize the number of eyes in groups. Eyes were divided into seven groups, each comprising seven eyes: unwounded control (Group 1), epithelial scrape (Group 2; aqueous humor samples taken at the 4th hour), superficial PRK (Group 3; samples taken at the 4th hour), deep PRK (Group 4; samples taken at the 4th hour), epithelial scrape (Group 5; samples taken at the 24th hour), superficial PRK (Group 6; samples taken at the 24th hour), and deep PRK (Group 7; samples taken at the 24th hour). The corneal epithelium was mechanically removed in surgical groups. The authors performed superficial corneal ablation (59 microm) in Groups 3 and 6 and deep corneal ablation (99 microm) in Groups 4 and 7. Aqueous humor samples were taken at the 4th hour (Groups 2-4) or 24th hour (Groups 5-7) after corneal surgeries. NO measurements were performed indirectly by using the Griess reaction with a spectrophotometer. RESULTS: Aqueous humor NO levels 4 hours after corneal surgery were statistically significantly lower than the control group (p<0.05). However, there was no difference among the surgical groups at the 4th hour (p>0.05). At the 24th hour, the deep PRK group had significantly lower NO levels than both the control group and Groups 5 and 6 (p<0.05). NO levels were normalized at the 24th hour in epithelial scrape and superficial PRK groups (p>0.05) but remained stable at lower levels in deep PRK groups (p<0.05). CONCLUSIONS: Corneal surgery caused low NO levels in aqueous humor 4 hours after surgery. However, 24 hours after surgery, NO levels normalized following epithelial scrape and superficial PRK and were stable at lower levels in the deep PRK group. Complications of deep PRK application are possibly induced by low NO existence in the aqueous humor. PMID- 15134106 TI - Effect of cataract surgery and foldable intraocular lens implantation on retinal nerve fiber layer as measured by scanning laser polarimetry with variable corneal compensator. AB - PURPOSE: Scanning laser polarimetry (SLP) enables the measurement of retinal nerve fiber layer (RNFL) thickness in vivo. As SLP measurements can be influenced by several conditions such as corneal birefringence, the new version GDx Access is implemented by the variable corneal compensator (VCC). The aim of this study was to evaluate the influence of cataract extraction and foldable intraocular lens (IOL) implantation on SLP parameters measured by GDx Access. METHODS: Sixty eight eyes of 68 patients undergoing phacoemulsification with foldable IOL implantation (silicone and acrylic) were examined by SLP before and after surgery. SLP was performed using GDx Access (with VCC). Mean values for SLP parameters were compared before and after 30 days from surgery with paired two tailed Student's t-test. RESULTS: No statistically significant differences were found between SLP parameters before and after cataract surgery, and regardless of the type of IOL implanted. CONCLUSIONS: Phacoemulsification and foldable IOL implantation do not influence RNFL parameters, as measured by GDx Access. PMID- 15134107 TI - Lidocaine-assisted xylocaine jelly anesthesia versus one quadrant sub-Tenon infiltration for self-sealing sclerocorneal incision routine phacoemulsification. AB - PURPOSE: To compare the effect of xylocaine jelly and intracameral lidocaine with one quadrant instant sub-Tenon infiltration for self-sealing sclerocorneal phacoemulsification. METHODS: One hundred patients were enrolled into a prospective randomized study, receiving either a combination of topical 2% xylocaine jelly and 0.5 ml of intracameral 1% lidocaine or sub-Tenon infiltration with 2 ml of 2% xylocaine on the operating table. All patients underwent a standard divide and conquer phacoemulsification procedure through a superior sclerocorneal frown incision followed by implantation of a polymethylmethacrylate intraocular lens. Intraoperative pain was indicated by the patient by squeezing the bedside nurse's hand, who allocated it to particular stages of surgery on a chart. After surgery, patients assessed the pain experienced using a 10-unit visual analogue scale. RESULTS: Pain was indicated on 31 occasions during the operation in the sub-Tenon group (mainly the injection itself) and 67 times in the topical group. The median overall subjective pain score was 3 in the jelly group and 0 in the sub-Tenon. Five eyes (10%) had to be converted to sub-Tenon during the surgery because of intolerable pain. CONCLUSIONS: Whereas lidocaine supported xylocaine jelly anesthesia provided acceptable analgesia for 90% of patients operated, sub-Tenon anesthesia proved to deliver better intraoperative comfort in all patients receiving sclerocorneal incision cataract surgery. PMID- 15134108 TI - Correlating intraocular pressure, blood pressure, and heart rate changes after jogging. AB - PURPOSE: To examine the effects of jogging on intraocular pressure (IOP), blood pressure (BP), and heart rate (HR). METHODS: Twenty-nine healthy individuals-25 athletes and 4 untrained-were studied. IOP, systolic and diastolic BP, and HR were measured before and just after 20 minutes of jogging (submaximal--70%- aerobic exercise). RESULTS: IOP decreased after jogging. Only three individuals had unchanged IOP in one eye and one individual in both eyes. The IOP decrease (1 to 8 mmHg) was statistically significant (p<0.001). BP increased after jogging (systolic: 0 to 60 mmHg, statistically significant changes, p<0.001; diastolic: 0 to 15 mmHg, statistically significant changes, p<0.001). HR increased as well (15 to 80 pulses/min, statistically significant changes, p<0.001). However, there were individuals who presented a significant decrease of IOP and a mild BP rise and vice versa, and also individuals with mild IOP decrease and significant HR change and vice versa. The statistical analysis clearly showed that there are no linear quantitative correlations between BP or HR changes and IOP changes. CONCLUSIONS: IOP decreases after jogging. Changes in BP and HR values have no linear quantitative correlation with IOP decrease. PMID- 15134109 TI - Subretinal choroidal neovascularization associated with choroidal nevus. AB - PURPOSE: Evaluation of a large series of choroidal nevi inducing the formation of a neovascular membrane in order to more clearly define the clinical presentation and to evaluate the efficacy of various treatment options. METHOD: Retrospective study of 22 clinical cases. RESULTS: All nevi were situated in the posterior choroid. They had a mean diameter of 3.8 mm and a mean thickness of 1.4 mm. Neovascular membranes were classic in all cases, extrafoveal in 13 cases (59%), and subfoveal in 9 cases (41%). A serous retinal detachment was present in every case, hemorrhages were present in 13 cases (59%), and lipid deposits were present in 16 cases (73%). All extrafoveal neovascular membranes were successfully treated by thermal laser photocoagulation. Initial visual acuity was 0.1 in three cases, 0.2-0.4 in five cases, 0.5-0.8 in four cases, and 1.0 or more in two cases. Final visual acuity was 0.1 in one case, 0.2-0.4 in one case, 0.5-0.8 in four cases, and 1.0 or more in seven cases. Five subfoveal neovascular membranes were treated either by thermal laser, photodynamic therapy, or irradiation. No treatment was applied in four cases and in one of these cases, spontaneous resolution of the neovascular membrane was observed. No growth of the pigmented tumor was observed with a mean follow-up of 4.8 years. CONCLUSIONS: Proliferation of a neovascular membrane on the surface of a pigmented choroidal tumor is a rare complication and is considered to be a relative indicator of a benign nature of the lesion. In the authors' experience, neovascular membranes are extrafoveal in more than half of cases and are accessible to laser photocoagulation. In contrast, the various modalities used to treat subfoveal neovascular membrane were ineffective and functional prognosis was unfavorable in these cases. PMID- 15134110 TI - Ultrasound biomicroscopy findings in peripheral vitreoretinal toxocariasis. AB - PURPOSE: To describe the morphologic alterations in ultrasound biomicroscopy (UBM) present in peripheral vitreoretinal toxocariasis. METHODS: An observational prospective study of case series. Fifteen eyes of 15 patients with clinical and laboratory diagnosis of peripheral vitreoretinal toxocariasis were enrolled. The patients were submitted to UBM examination of the region corresponding to the pars plana of the affected eye. RESULTS: The most common morphologic alterations found by UBM in patients with peripheral vitreoretinal toxocariasis were as follows: vitreal membranes (13 cases), toxocara granuloma (11 cases), and pseudocysts (8 cases). Other less frequent findings were thickening of the ciliary body (6 cases), cystic formation (2 cases), peripheral retinal detachment (2 cases), rectification of the iris root (1 case), and posterior synechiae (1 case). CONCLUSIONS: UBM allows detection of well-defined morphologic alterations associated with peripheral vitreoretinal toxocariasis, being useful to reinforce the clinical diagnosis. PMID- 15134111 TI - Vitrectomy results for diffuse diabetic macular edema with and without inner limiting membrane removal. AB - PURPOSE: To determine whether vitrectomy for diffuse diabetic macular edema with and without internal limiting membrane (ILM) peeling is equally effective in reducing edema. METHODS: The authors retrospectively analyzed the surgical outcomes in 73 eyes of 52 patients with diffuse diabetic macular edema. Eighteen eyes (Group A) underwent three-port pars plana vitrectomy with posterior hyaloid membrane (PHM) removal, while 55 eyes (Group B) had pars plana vitrectomy with additional ILM peeling after PHM removal. RESULTS: Intraoperatively, the posterior hyaloid was found to be attached to the macula in all eyes. In Group A, macular edema resolved completely in 8 eyes (44.4%) with improvement of visual acuity (VA). In Group B, VA improved in 38 eyes (69.1%) with complete resolution of edema. The results of this study indicated that vitrectomy effectively reduced macular edema but eyes with ILM peeling (Group B) presented better results than those without ILM peeling. Another important factor related to the outcome seems to be the level of glycosylated hemoglobin (HbA1c). CONCLUSIONS: In eyes with diffuse diabetic macular edema vitrectomy seems to be effective, but additional ILM peeling presented better results. PMID- 15134112 TI - Ocular injuries secondary to motor vehicle accidents. AB - PURPOSE: Ocular trauma is one of the main causes of visual reduction or loss, particularly in the younger population. METHODS: In this prospective study the authors included 67 consecutive patients with ocular trauma secondary to motor vehicle accidents who were hospitalized in the Athens University Eye Clinic from September 1993 to December 1996. The mean follow-up time was 31 months, the mean age was 31.7 years, and the ratio between men and women was 2.7:1. RESULTS: Thirty-two of the accidents (47.76%) took place in populated areas. Among the 67 injured persons, 58 (86.56%) were car passengers, 8 (11.95%) were on motorcycles, and 1 (1.49%) was a pedestrian. Only 3 (5.2%) of the 58 persons injured inside automobiles used safety belts and none of the motorcyclists used crash helmets during the accidents. Fifty-three (79.1%) ocular traumas were penetrating in nature, with glass fragments being the main cause in 36 of them (67.9%). Among the 53 injured persons experiencing penetrating ocular trauma, 49 had a follow-up time of more than 6 months. Twenty of them (40.8%) underwent one surgical procedure, 22 (44.9%) were submitted to two surgical procedures, and the remaining 7 persons (14.3%) needed three or more operations. Eighteen (36.7%) of the 49 patients with penetrating ocular trauma and with 6 months follow-up had a final visual acuity of less than 1/20, 21 (42.9%) had a visual acuity of more than 5/10, and 3 (6.1%) underwent enucleation. Among the 67 patients, 61 had a follow-up time of more than 6 months, regardless of their history of penetrating ocular trauma. Eighteen of them (29.5%) had a final visual acuity of less than 1/20, 8 (13.1%) had a visual acuity between 2/10 and 4/10, and 22 (52.5%) had a visual acuity of more than 5/10. CONCLUSIONS: Because motor vehicle accidents can cause severe ocular trauma, it would be helpful for drivers to be more careful and aware of motor vehicle regulations. There seems to be a great need of enforcement of seatbelt laws in Greece. PMID- 15134113 TI - Scleral fixation of a posterior chamber intraocular lens in a patient with two dislocated lens implants. AB - PURPOSE: To report the clinical findings and management of a case of two consecutive intraocular lenses (IOLs) dislocated into the vitreous cavity after complicated cataract surgery. METHODS: Review of clinical findings and treatment. RESULTS: A 69-year-old man sought treatment for posterior chamber IOL dislocation. The patient reported a significant loss of visual acuity in the right eye for 4 months (best-corrected visual acuity (BCVA) 20/200 in the affected eye). In the vitreous cavity two dislocated IOLs were found, complicated by a cystoid macula edema. Surgery was planned and the two IOLs were removed from the vitreous cavity. Aphakia was then corrected by means of a scleral fixated posterior chamber lens. Three months after surgery, BCVA was 20/40. CONCLUSIONS: In the absence of randomized controlled clinical trials evaluating treatment options, preoperative patient work-up should be as accurate as possible in order to reduce the risk of intraoperative and postoperative complications. In this patient, removing the two IOLs and placing a sutured-fixated posterior chamber IOL allowed resolution of the cystoid macular edema. PMID- 15134114 TI - Choroidal effusion and hypotony caused by severe anterior lens capsule contraction following cataract surgery. AB - PURPOSE: The case report describes a case of severe anterior capsular contraction associated with choroidal effusion. CASE REPORT: An 81 year old female with primary open angle glaucoma underwent routine phacoemulsification cataract surgery. Eight weeks following surgery the anterior capsule opening had reduced to 3 mm in size. Intraocular pressure was found to be 4 mmHg and B scan ultrasound revealed a large choroidal effusion. Anterior capsulotomy with Nd:YAG laser was performed. At review, two weeks later, the choroidal effusion had resolved and visual acuity had recovered. DISCUSSION: The Nd:YAG laser radial relaxing capsulotomies helped relieve the capsular contraction and associated traction on the ciliary body. PMID- 15134115 TI - Pigmented congenital vitreous cyst. AB - PURPOSE: To describe the clinical findings in a case with a typical pigmented congenital vitreous cyst and to discuss its differential diagnosis. METHODS/CASE REPORT: A 24-year-old woman complaining of floaters and blurring of vision during eye movements in the right eye was examined. RESULTS: A diagnosis of congenital vitreous cyst was made after detailed vitreoretinal examination, B-scan ultrasonography (eye and abdomen), computerized tomography of brain, chest radiogram, serologic tests for Echinococcosis and Cysticercosis in the serum, and complete blood count for eosinophilia. Clinical presentation and the patient's complaints remained unchanged during 3 years of follow-up. CONCLUSIONS: Congenital vitreous cysts are benign lesions. Differential diagnosis from acquired cysts requires careful clinical examination and appropriate laboratory tests. PMID- 15134117 TI - Duplication of the optic disc: true or pseudo? A coloboma or not a coloboma? AB - PURPOSE: To present an unusual case of optic disc pseudoduplication with colobomata. METHODS: Clinical evaluation, fundus photography and literature review. RESULTS: Optic disc duplication is a rare clinical entity. CONCLUSIONS: We report what we believe to be the first case of pseudoduplication of the optic disc with coexistent bilateral optic disc colobomata. PMID- 15134116 TI - Choroidal neovascularization in myopic eyes after phakic refractive lens and iris claw lens implantation. AB - PURPOSE: To describe the appearance of choroidal neovascularization (CNV) in two myopic patients after implantation of a phakic refractive lens (PRL) and an iris claw lens. METHODS: A PRL was implanted in the left eye of a 35-year-old myopic man. Five weeks later, he reported decreased best-corrected visual acuity (BCVA). Fluorescein angiography revealed juxtafoveal CNV Photodynamic therapy with Visudyne (PDT) was successfully performed, achieving closure of the membrane. BCVA was 20/40 3 months afterwards. An iris-claw lens was implanted in the left eye of a 24-year-old myopic man. BCVA after surgery was 20/40 (SE -0.75). Three years later subfoveal CNV was diagnosed. PDT was performed, achieving complete closure of CNV BCVA was 20/100 3 months after treatment. CONCLUSIONS: The appearance of CNV in myopic eyes corrected by phakic intraocular lens implantation is a possible complication that must be considered. PMID- 15134118 TI - Retinal detachment in focal dermal hypoplasia. AB - PURPOSE: Focal dermal hypoplasia is a systemic disease that includes well recognized ocular abnormalities. Retinal detachment has not previously been reported as a part of this syndrome. AIM: To report a case of focal dermal hypoplasia with an associated retinal detachment. METHODS: Single case report of a child with a colobomatous retinal detachment and the focal dermal hypoplasia syndrome. CONCLUSIONS: Retinal detachments may occur as part of the focal dermal hypoplasia syndrome. Expanding knowledge of this syndrome may help ophthalmologists diagnose this rare condition. PMID- 15134119 TI - Understanding the patient's perspective on rapid and routine HIV testing in an inner-city urgent care center. AB - The purpose of this study was to explore patient perspectives of rapid and routine HIV-testing in an urgent care center at an urban public hospital. We conducted structured focus groups during a clinical trial comparing routinely offered rapid HIV-testing, routinely offered enzyme immunoassay (EIA) testing, and conventional EIA testing. Participants of the six focus groups were 89% African American, 60% uninsured, and had a low educational status. Four independent coders analyzed the data using iterative content analysis. Rapid testing was preferred to EIA testing because it reduced the need for a return visit and stress of waiting for test results, though there were concerns about accuracy. Participants supported routinely offering testing, but there were concerns about privacy and cost. Fear and stigma were common reasons for refusing testing and not returning for results. Distrust and misconceptions about HIV, particularly regarding the importance of testing, were very common. PMID- 15134120 TI - Development of a measure of barriers to HIV testing among individuals at high risk. AB - Rates of HIV antibody testing remain at approximately 45% of the general population. To more effectively design interventions to increase testing, comprehensive information is needed to understand the barriers to HIV testing. A measure of barriers to HIV testing was developed using the major barriers identified in the literature on barriers to health care utilization (Melnyk, 1988), and tested with a diverse group of individuals at high risk for HIV, including heterosexuals, men who have sex with men, injected drug users, and sex workers. An exploratory factor analysis indicated that the factor structure was replicated over 2 years of data collection. Three factors--Structural Barriers, Fatalism/Confidentiality Concerns, and Fear--emerged for both years. The reliabilities ranged from .75 to .87, indicating moderate to high internal consistency. PMID- 15134121 TI - HIV testing in a resource-poor urban emergency department. AB - In an effort to help fight the HIV epidemic, we offered standard HIV testing to high-risk and symptomatic patients attending an urban emergency department (ED) located in a high-prevalence area. We assessed rates of consent, HIV infection, and linkage into care by demographic and risk characteristics. Consent rates were consistently greater than 50% across the various demographics and risk groups. The HIV prevalence rate was 3% and ranged from 0% to 50% across groups. Of those eligible to enter care 69% attended their first infectious disease clinic appointment. One recommendation to increase the number of people who are aware of their serostatus is to offer HIV testing in underutilized venues, such as an urban ED. The results of this project offer suggestions for future pursuits. PMID- 15134122 TI - Development of an opinion leader-led HIV prevention intervention among alcohol users in Chennai, India. AB - In 1999, we began a community-based randomized controlled prevention trial in Chennai, which aims to test the efficacy of HIV prevention messages disseminated through members of an individual's social group called community popular opinion leaders, or CPOLs. We targeted patrons of 100 bars or wine shops in the city of Chennai, India. In this article we report on the process of development of an HIV prevention intervention for wine shop patrons. First, we conducted detailed ethnography to understand social norms and CPOL and social network characteristics, including 41 in-depth interviews among wine shop patrons and gatekeepers. Second, we tailored a generic HIV education training manual to appropriately address the needs of Chennai wine shop patrons. Field-testing involved 16 focus groups with wine shop patrons and 12 sessions of participant observations in wine shops. Finally, we piloted the intervention to determine the appropriateness of the training program and its content among wine shop patrons. Our ethnographic data indicated that wine shops are a common meeting place for men. We were able to identify CPOLs influential in these settings and train them to deliver appropriate prevention messages to their close friends and associates. We found that HIV prevention messages in this population need to dispel misperceptions about HIV transmission, provide strategies and skills to adopt and sustain condom use, and target the role of alcohol in sexual behavior. We outline specific lessons we learned in intervention development in this population. PMID- 15134123 TI - Prevention myths and HIV risk reduction by active drug users. AB - Drug injectors and crack users (526) in South Florida responded to a survey questionnaire that was designed to examine belief in the effectiveness of various strategies, other than condom use, employed to reduce personal risk of contracting HIV during sexual acts. Each strategy was believed to be effective by at least one quarter of the study participants. Factor analysis was used to group these strategies. Subsequent multivariate analysis indicated that the participants who believed in the effectiveness of HIV prevention strategies other than condom use were also less likely to report using condoms. These findings highlight the need for prevention interventions to elicit prevention myths and the full range of risk reduction strategies practiced. PMID- 15134124 TI - What really works? An exploratory study of condom negotiation strategies. AB - Verbal-direct strategies are assumed to be the most effective strategies in negotiating condom use. Both cultural and gender differences in communication styles suggest that individuals may negotiate condoms in ways that are not exclusively verbal and direct. This study examined the use of other forms of condom negotiations by developing an exploratory scale that distinguished strategies on how verbal and direct they were (i.e., verbal-direct, verbal indirect, nonverbal-direct, nonverbal-indirect). The study compared the use of negotiation strategies among Asian and White American students at a northern California university. Results indicated that although direct strategies (verbal and nonverbal) were more frequently used, condom users also employed indirect strategies (verbal and nonverbal) to negotiate condom use. Moreover, Asians used verbal-indirect strategies more than Whites. Women used nonverbal-indirect strategies more than men. HIV preventions seeking to be culturally sensitive to Asians and women may benefit from incorporating these strategies into their interventions. PMID- 15134125 TI - Gender differences in condom-related behaviors and attitudes among Mexican adolescents living on the U.S.-Mexico border. AB - Adolescents are at increasing risk for HIV infection in Mexico. Research on gender differences in risk behaviors and determinants is needed to develop effective HIV prevention interventions targeting Mexican adolescents. This study examined gender differences in the likelihood of unprotected sex and theoretical correlates among high school students in the border city of Tijuana. Three hundred seventy high-school students completed a face-to-face interview and a self-administered survey. Differences in sexual initiation, condom use, intentions to use condoms in the future, and attitudes towards condoms in this population were assessed. Although male students initiated sexual practices earlier than females, females were more likely to have unprotected sex. Females perceived themselves as more likely to avoid unprotected sex in the future and held more favorable attitudes about condoms. The results suggest that stereotypical gender roles and communication barriers place Tijuana female high school students at higher risk for HIV infection than their male peers. PMID- 15134126 TI - Behavioral problems and scholastic adjustment among Bedouin-Arab children from polygamous and monogamous marital family structures: some developmental considerations. AB - Families in the Bedouin-Arab community in Israel are characterized by monogamous and polygamous marriages. Such diversity in family structure occurs in other parts of the world, yet scant empirical evidence exists to refute or to support the claim that polygamous family structure can be a risk factor for children's school maladjustment and negative developmental outcomes. The purpose of the current study was to test this claim. Participants were 255 3rd-grade children from the Negev Bedouin community in Israel. One hundred fifty-three children came from monogamous families that were characterized by 1 wife (i.e., 1-wife families), and 102 children came from polygamous families consisting of 2 wives (i.e., 2-wife families). Teachers completed the Teacher's Report Form of the Child Behavior Checklist (T. M. Achenback, 1991b). A series of logistic regression analyses, after adjusting for maternal education level, revealed that 2-wife children tended to have higher levels of externalizing problems in general and higher levels of attention problems in particular than did their 1-wife counterparts. Also, 2-wife children had higher rates of school absenteeism and lower levels of overall academic achievement than did 1-wife children. Implications for the Bedouin society are discussed. PMID- 15134127 TI - Two facets of self-knowledge: cross-cultural development of measures in Iran and the United States. AB - Self-knowledge is an ideal not only within psychological theory and practice but also within the religious and philosophical foundations of many cultures. In 6 studies conducted in Iran and the United States, the authors sought to construct and to validate scales for measuring two facets of self-knowledge. Experiential self-knowledge was defined as an ongoing sensitivity to the self in the present. Reflective self-knowledge was described in terms of personal efforts to integrate experience within self-schemas developed in the past. Thirteen-item experiential self-knowledge and reflective self-knowledge scales were created by the authors using samples of Iranian and American university students. A confirmatory factor analysis verified this 2-factor structure in a second study, and these results were replicated in a 3rd study. Correlations with a broad array of self-report variables established the two scales as valid measures of adjustment. Both displayed adequate test-retest reliability. Correlations with peer reports suggested that the two factors had behavioral implications in both cultures. Reflective self-knowledge proved to be as important as educational abilities in predicting the academic performance of Americans who were motivated to attend class. Experiential self-knowledge and reflective self-knowledge also interacted to predict better grades. In short, the experiential self-knowledge and reflective self-knowledge scales operationalized cross-cultural personality processes that deserve additional research attention. PMID- 15134128 TI - Facial appearance and judgments of credibility: the effects of facial babyishness and age on statement credibility. AB - Researchers have found that facial appearance influences social judgments. For example, evidence has shown that facial babyishness and age affect perceivers' impressions of the stimulus person's veracity. In this experiment, the researchers examined whether these variables also influenced the credibility attributed to written statements purportedly made by these people in addition to several topics of interest in deception-detection research. Undergraduates (N = 270) were presented babyfaced or mature-faced photographs that depicted a child, an adult, or an older individual, in addition to a written truthful or deceptive statement purportedly made by the person in the photograph. Results showed that, as predicted, when the statements were accompanied by babyfaced pictures, participants tended to judge them as truthful, but only if the pictures did not depict children. Also, when the statements were accompanied by childen's pictures, participants tended to judge them as deceptive, but only if the pictures depicted a babyish face. Overall detection accuracy was close to chance and did not correlate with either judgmental confidence or with the respondents' estimated lie-detection accuracy. However, confidence and estimated ability were significantly correlated. Also, more confidence was placed in judgments of truthfulness than in judgments of deceptiveness. Respondents' truth bias and the existence of a veracity effect in the diverse experimental conditions were examined as well. PMID- 15134129 TI - Mass use of insecticide-treated bednets in malaria endemic poor countries: public health concerns and remedies. AB - Epidemiological evidence arising mainly from studies undertaken in sub-Saharan Africa suggests that sleeping under insecticide-treated mosquito nets is a cost effective and efficacious method of controlling malaria. For this reason, promotion of use of insecticide-treated nets (ITNs) has become a key malaria control strategy. In 1999, the United Nations Children's Fund (UNICEF) and WHO set the goal of providing 32 million nets and 320 million net treatments a year for the next 10 years to protect 80% of African households against malaria. All pesticides are toxic by nature and are associated with adverse health risks that depend on the toxicity of each chemical, as well as the type and degree of exposure. Thus, massive scale-up of use of ITNs in malaria endemic poor countries can be expected to present tangible risks to health, especially where the insecticides for net treatment and re-treatment are handled mostly by untrained persons in uncontrolled settings. This paper examines potential health risks of mass use of ITNs in malaria endemic poor countries and calls for the implementation of strategies to minimize potential risks through careful selection of products, appropriate labeling (including labeling in the local languages of the user communities), pesticide safety education of the public and training of health personnel, and active monitoring of adverse health effects to document actual and potential hazards, and to facilitate planning of mitigation efforts. PMID- 15134130 TI - Circumstances leading to the formulation and implementation of a new TB control program in Israel: a case study in public health and policy. AB - We describe how Israel effected new tuberculosis programs and policies beginning in the early 1990s. We explain how the epidemiology of the disease, particular events, and a small number of people influenced the creation of new policy. We believe that this story may be useful to other program managers and policymakers interested in changing course. PMID- 15134131 TI - What's in a name? Policy transfer in Mozambique: DOTS for tuberculosis and syndromic management for sexually transmitted infections. AB - A common assumption is that international health policies are imposed on developing countries, owing to their high level of dependence on international aid. In reality, the process is likely to be complex. Drawing on analytical frameworks developed to study policy transfer between jurisdictions, this paper explores how far two globally promoted infectious disease policies (DOTS for tuberculosis and syndromic management for sexually transmitted infections) were voluntarily or coercively transferred in one particular setting, Mozambique. The paper suggests that guidelines emanating from these policies were not imposed, but evolved in the 1980s through technical networks of national and international experts. Further, that it was experience at the country level that fed into the globally promoted policies of the 1990s. By the time the policies were transferred by WHO and other international organisations to developing countries in the 1990s, Mozambique had already adopted their guidelines for good practice. PMID- 15134132 TI - What's in a policy context? PMID- 15134133 TI - Public attitudes about underage drinking policies: results from a national survey. AB - We conducted a national telephone survey of 900 adults in the United States to examine the attitudes of the adult public regarding underage drinking and a series of alcohol control policies aimed at reducing it. Three versions of the survey instrument were administered, each to one-third of the sample, with the versions varying in the stipulations of the policy options. Results showed high levels of public support for most of the alcohol control policies, with relatively lower support for those that would result in restrictions on adults' access to alcohol. Respondents' support of the policy options was significantly related to their sociodemographic and attitudinal characteristics, such as sex, age, drinking frequency, and level of concern about underage drinking. The findings provide important guidelines to policymakers interested in garnering support for policies aimed at curtailing underage drinking. PMID- 15134134 TI - Public health interactions with the public: can quality be assured? "This call may be monitored for quality assurance purposes". AB - I describe what happened when a citizen called her state health department "after hours" to report possible contamination of food at a commercial establishment. The call, inadequately handled by the health department, illustrates the need to assure quality of the public health responses to calls on a 24 hour, 7-day week basis. I examine possible reasons for the poor response. Health departments should consider training for those assigned to handle phone calls from the public, such as that provided to poison control center personnel. In addition, a quality assurance program should routinely assess the adequacy of public health responses. As medical care is available on an emergency basis, 24 hours a day, 7 days a week, emergency services for prevention of illness in the population should be available to the public at all times with the response provided by appropriately trained individuals. PMID- 15134135 TI - Lessons from the margins of globalization: appreciating the Cuban health paradox. AB - It is widely recognized that Cuba, despite poor economic performance, has achieved and sustained health indices comparable to those in developed countries- the Cuban Paradox. There has been, however, remarkably little scholarship evaluating how this has been accomplished, especially during a period of extreme economic hardship. Cuba's exclusion from the mainstream of "globalization," moreover, allows us to gain insights into the population health impact of policies that have accompanied globalization. Cuba's experience challenges the conventional assumption that generating wealth is the fundamental precondition for improving health. As peoples around the world search for cost-effective ways to improve well-being, they might want to learn how alternative public policy approaches, such as those used in Cuba, may be effective. We therefore reviewed the literature on the health-wealth relationship in this globalizing era; then systematically examined public policy in Cuba, not only for health services (financing, vertical and horizontal integration, prevention and primary-care focus, inter-sectoral linkages, etc.) but for non-medical determinants of health as well. These included education, housing, nutrition, employment, etc. plus the community mobilization and social cohesion that the Cuban system has generated. It appears that the active implementation of public policy affecting a wide variety of health determinants explains the Cuban paradox, and that the international community can learn from Cuba's experience. The prospect for healthy public policy can thus exist within, rather than only on the margins of globalization. The importance of monitoring how Cuba sustains such policies as it faces growing challenges in this globalizing era is increasingly worth observing. PMID- 15134136 TI - Phospholipids and oxophospholipids in atherosclerotic plaques at different stages of plaque development. AB - We identified and quantified the hydroperoxides, hydroxides, epoxides, isoprostanes, and core aldehydes of the major phospholipids as the main components of the oxophospholipids (a total of 5-25 pmol/micromol phosphatidylcholine) in a comparative study of human atheroma from selected stages of lesion development. The developmental stages examined included fatty streak, fibrous plaque, necrotic core, and calcified tissue. The lipid analyses were performed by normal-phase HPLC with on-line electrospray MS using conventional total lipid extracts. There was great variability in the proportions of the various oxidation products and a lack of a general trend. Specifically, the early oxidation products (hydroperoxides and epoxides) of the glycerophosphocholines were found at the advanced stages of the plaques in nearly the same relative abundance as the more advanced oxidation products (core aldehydes and acids). The anticipated linear accumulation of the more stable oxidation products with progressive development of the atherosclerotic plaque was not apparent. It is therefore suggested that lipid infiltration and/or local peroxidation is a continuous process characterized by the formation and destruction of both early and advanced products of lipid oxidation at all times. The process of lipid deposition appears to have been subject to both enzymatic and chemical modification of the normal tissue lipids. Clearly, the appearance of new and disproportionate old lipid species excludes randomness in any accumulation of oxidized LDL lipids in atheroma. PMID- 15134137 TI - Lactational changes in the fatty acid composition of human milk gangliosides. AB - The objectives of this work were to study the FA composition of milk gangliosides, as well as to gain further insight into the characterization of human milk gangliosides. The potential capacity of human milk gangliosides to adhere to human enterotoxigenic Escherichia coli (ETEC-strains) was also studied. Human milk gangliosides were isolated and identified by high-performance TLC or immunoassay. The latter also was used to assay bacterial adhesion. The FA composition of gangliosides was studied by GC. The presence of O-acetyl GD3 (Neu5,9Ac2alpha2-8 NeuAcalpha2-3Galbeta1-4GlcCer) and trace amounts of GM1 [Galgamma1]3-3GalNAcgamma1,-3(Neualpha2-3)Galbeta1-4GlcCerl in human milk was confirmed. Medium-chain FA were almost absent in colostrum, whereas in the subsequent stages they rose to 20%. The levels of long-chain FA decreased after colostrum. With respect to the degree of saturation, gangliosides from colostrum were richer in monounsaturated FA than gangliosides synthesized during the rest of the lactation period, opposite to the pattern for PUFA. A human-ETEC colonization factor antigen II-expressing strain showed binding capacity to human milk GM3 (NeuAcalpha2-3Gal[1-4GlcCer). New data on human milk gangliosides have been gathered. A thorough knowledge of their composition is needed since they may have important biological implications in regard to newborns' defense against infection. PMID- 15134138 TI - Blood phospholipid fatty acid analysis of adults with and without attention deficit/hyperactivity disorder. AB - Several psychiatric disorders, including juvenile Attention Deficit/Hyperactivity Disorder (ADHD), have been associated with abnormalities of certain long-chain PUFA (LCPUFA). Despite this reported association, the FA levels of patients with the adult form of ADHD have not previously been evaluated. In this study we measured the total blood phospholipid FA concentrations in 35 control subjects and 37 adults with ADHD symptoms to determine whether adults with ADHD symptoms would show abnormalities of FA relative to control subjects. In the serum phospholipids, adults with ADHD symptoms had significantly lower levels of total saturated, total polyunsaturated, and total omega-6 (n-6) FA, as well as the omega-3 (n-3) LCPUFA DHA (22:6n-3), and significantly higher levels of total monounsaturated FA and the n-3 LCPUFA docosapentaenoic acid (22:5n-3). In the erythrocyte membrane phospholipids, adults with ADHD symptoms had significantly lower levels of total PUFA, total n-3 FA, and DHA, and significantly higher levels of total saturated FA. Neither serum nor erythrocyte membrane phospholipid DHA was related to ADHD symptom severity (as assessed by the Amen questionnaire) in ADHD subjects. Although the exact cause of these variations is unknown, both environmental and genetic factors may be involved. PMID- 15134139 TI - Dietary linoleic acid-induced hypercholesterolemia and accumulation of very light HDL in steers. AB - This experiment was designed to study the effects in fattening steers of n-6 PUFA supplementation on the plasma distribution and chemical composition of major lipoproteins (TG-rich lipoproteins: d < 1.006 g/mL; intermediate density lipoproteins + LDL: 1.019 < d < 1.060 g/mL; light HDL: 1.060 < d < 1.091 g/mL; and heavy HDL: 1.091 < d < 1.180 g/mL). For a period of 70 d, animals [454 +/- 20 d; 528 +/- 36 kg (mean +/- SD)] were given a control diet (diet C, n = 6) consisting of hay and concentrate mixture (54 and 46% of diet dry matter, respectively) or the same diet supplemented with sunflower oil (4% of dry matter), given either as crushed seeds (diet S, n = 6) or as free oil continuously infused into the duodenum through a chronic canula to avoid ruminal PUFA hydrogenation (diet O, n = 6). Plasma lipids increased in steers given diet S (x1.4, P < 0.05) and diet O (x2.3, P < 0.05), leading to hyperphospholipemia and hypercholesterolemia. With diet S, hypercholesterolemia was associated with higher levels of light (x1.4, P < 0.05) and heavy HDL (x1.3, NS). With diet O, it was linked to higher levels of light HDL (x1.8, P < 0.005) and to very light HDL accumulation within density limits of 1.019 to 1.060 g/mL, as demonstrated by the apolipoprotein A-I profile. Diet O favored incorporation of 18:2n-6 into polar (x2.2, P < 0.05) and neutral lipids (x1.5 to x8, P < 0.05) at the expense of SFA, MUFA, and n-3 PUFA. Thus, protection of dietary PUFA against ruminal hydrogenation allowed them to accumulate in plasma lipoproteins, but the effects of hypercholesterolemia on animal health linked to very light HDL accumulation remain to be elucidated. PMID- 15134140 TI - Contrasting effects of t10,c12- and c9,t11-conjugated linoleic acid isomers on the fatty acid profiles of mouse liver lipids. AB - The purpose of this study was to examine the effects of two purified isomers of CLA (c9,t11-CLA and t10,c12-CLA) on the weights and FA compositions of hepatic TG, phospholipids, cholesterol esters, and FFA. Eight-week-old female mice (n = 6/group) were fed either a control diet or diets supplemented with 0.5% c9,t11 CLA or t10,c12-CLA isomers for 8 wk. Weights of liver total lipids and those of individual lipid fractions did not differ between the control and the c9,t11-CLA groups. Livers from animals fed the t10, c12-CLA diet contained four times more lipids than those of the control group; this was mainly due to an increase in the TG fractions (fivefold), but cholesterol (threefold), cholesterol esters (threefold), and FFA (twofold) were also significantly increased. Although c9,t11 CLA did not significantly alter the weights of liver lipids when compared with the control group, its intake was associated with significant reductions in the weight percentage (wt% of total FAME) of 18:1n-9 and 18:1n-7 in the TG fraction and with significant increases in the weight percentage of 18:2n-6 in the TG, cholesterol ester, and phospholipid fractions. On the other hand, t10,c12-CLA intake was linked with a significant increase in the weight percentage of 18:1n-9 and a decrease in that of 18:2n-6 in all lipid fractions. These changes may be the result of alterations in the activity of delta9-desaturase (stearoyl CoA desaturase) and the enzymes involved in the metabolism of 18:2n-6. Thus, the two isomers differed not only in their effects on the weights of total liver lipids and lipid fractions but also on the FA profile of the lipid fractions. PMID- 15134142 TI - Beta-oxidation of 18:3n-3 in Atlantic salmon (Salmo salar L.) hepatocytes treated with different fatty acids. AB - To study whether Atlantic salmon beta-oxidation was affected by dietary FA composition, an in vitro study with primary hepatocytes was undertaken. Isolated hepatocyte cultures were stimulated with either 16:0, 18:1n-9, 18:2n-6, 18:3n-3, 20:5n-3, or 22:6n-3 in triplicate for 24 h. In addition, a control was included where no FA stimulation was performed, also in triplicate. After stimulation, radiolabeled [1-14C] 18:3n-3 was added and the cells were incubated for 2 h at 20 degrees C. The cells were then harvested, and radioactivity was determined in the acid-soluble part of the cells and medium, i.e., the end products of the beta oxidation pathway. Specific beta-oxidation activity was significantly higher in hepatocytes stimulated with 18:3n-3. Further, when taking into account the amount of radiolabeled [1-14C]18:3n-3 taken up by the cells--the relative amount of beta oxidized [1-14C]18:3n-3 of the total FA taken up by the hepatocytes-no significant differences were found. Thus, the regulation of beta-oxidation activity in the primary Atlantic salmon hepatocytes seems to be at the level of FA uptake and transport into the cell. This in vitro study shows that the catabolism processes in salmon hepatocytes are affected by the FA available and probably already regulated at the level of FA uptake. PMID- 15134141 TI - Modulation of HepG2 cell net apolipoprotein B secretion by the citrus polymethoxyflavone, tangeretin. AB - The purpose of the present study was to examine the role of tangeretin, a polymethoxylated flavone from citrus fruits, on the regulation of apolipoprotein B (apoB) and lipid metabolism in the human hepatoma cell-line HepG2. The marked reduction in apoB secretion observed in cells incubated with 72.8 microM tangeretin was rapid, apoB-specific, and partly reversible. The reduction also was observed under lipid-rich conditions and found to be insensitive to proteasomal degradation of nascent apoB. We followed our study by examining lipid synthesis and mass. A 24-h exposure of cells to 72.8 microM tangeretin decreased intracellular synthesis of cholesteryl esters, free cholesterol, and TAG by 82, 45, and 64%, respectively; tangeretin also reduced the mass of cellular TAG by 37%. The tangeretin-induced suppression of TAG synthesis and mass were associated with decreased activities of DAG acyltransferase (up to -39.0 +/- 3.0% vs. control) and microsomal triglyceride transfer protein (up to -35.5 +/- 2.5% vs. control). Tangeretin was also found to activate the peroxisome proliferator activated receptor, a transcription factor with a positive regulatory impact on FA oxidation and TAG availability (up to 36% increase vs. control). The data suggest that tangeretin modulates apoB-containing lipoprotein metabolism through multiple mechanisms. PMID- 15134143 TI - Decreased production of inflammatory mediators in human osteoarthritic chondrocytes by conjugated linoleic acids. AB - Osteoarthritic chondrocytes (OC) produce excessive prostaglandin E2 (PGE2) and nitric oxide (NO), which function as inflammation mediators in the pathogenesis of osteoarthritis (OA). This study examined the effect of CLA alone and in combination with other PUFA on the FA composition and the production of PGE2 and NO in OC cultures isolated from OA patients. Human OC were grown in monolayer and treated with one of the following PUFA treatments: CLA, CLA + arachidonic acid (CLA/AA), CLA + EPA (CLA/EPA), linoleic acid (LA), LA + AA (LA/AA), LA + EPA (LA/EPA), and ethanol (as a vehicle control) at 10 and 20 microM for 6 d. Supplementation of PUFA at 10 microM for 6 d did not introduce any cytotoxic effects or morphological changes in OC, whereas 20 microM resulted in apoptosis. Cultures of OC treated with CLA, CLA/AA, and CLA/EPA had higher concentrations of CLA isomers, and these isomers were not detected in other treatments. Supplementation of CLA or LA alone to the OC led to a lower PGE2 production compared to the control. Combination of CLA/EPA resulted in the lowest PGE2 production in cultured OC. OC cultures treated with CLA were lower in NO production than the control, whereas the LA/AA treatment demonstrated the lowest NO production. The fact that CLA alone or in combination with other PUFA modulated PGE2 and NO production in human OC cultures suggests that these 18:2 isomers may have the potential to influence OA pathogenesis. PMID- 15134144 TI - Synthesis and anticancer activities of fatty acid analogs of podophyllotoxin. AB - Derivatives of podophyllotoxin were prepared by coupling 10 FA with the C4-alpha hydroxy function of podophyllotoxin. The coupling reactions between FA and podophyllotoxin were carried out by dicyclohexylcarbodiimide in the presence of a catalytic amount of dimethylaminopyridine to produce quantitative yields of desired products. FA incorporated were the following: 10-hydroxydecanoic, 12 hydroxydodecanoic, 15-hydroxypentadecanoic, 16-hydroxyhexadecanoic, 12 hydroxyoctadec-Z-9-enoic, eicosa-Z-5,8,11,14-tetraenoic, eicosa-Z-8,11, 14 trienoic, eicosa-Z-11,14-dienoic, eicosa-Z-11-enoic, and eicosanoic acids. Spectroscopic studies confirmed the formation of the desired products. New molecules were investigated for their in vitro anticancer activity against a panel of human cancer cell lines including SK-MEL, KB, BT-549, SK-OV-3 (solid tumors), and HL-60 (human leukemia) cells. Most of the analogs were cytotoxic against cancerous cells, whereas no effect was observed against normal cells, unlike the parent compound podophyllotoxin, the use of which is limited due to its severe side effects. PMID- 15134145 TI - A spectroscopic study of the interaction of nigerloxin, a fungal metabolite, with serum albumin. AB - Nigerloxin [2-amido-3-hydroxy-6-methoxy-5-methyl-4-(prop-1'-enyl) benzoic acid], a fungal metabolite, is an inhibitor of lipoxygenase and aldose reductase with free radical-scavenging properties. The interaction of nigerloxin with bovine serum albumin (BSA) was investigated using fluorescence spectroscopy and circular dichroic measurements. The fluorescence of BSA was quenched following interaction with nigerloxin, and this property was used to generate a binding constant. The estimated association constant was 1.01 +/- 0.2 x 10(6) M(-1). Job's method of continuous variation indicated that nigerloxin formed a 1:1 +/- 0.1 complex with BSA. To understand the nature of the interaction, the variance in the association constant as a function of temperature in the range of 14-45 degrees C was used to calculate the thermodynamic parameters. The thermodynamic parameters at 27 degrees C derived from the mass action plot and van't Hoff's plot were as follows: deltaG = -8.2 +/- 0.1 kcal/mol, deltaH approximately equal to 0 kcal/mol, and deltaS = 27.5 +/- 0.4 cal/mol/K (where deltaG is free energy, deltaH is enthalpy, and deltaS is entropy). Increasing ionic strength did not favor interaction. Circular dichroic measurements revealed that the interaction of nigerloxin with BSA did not lead to changes in the secondary structure of the protein. The reversibility of the interaction verified by the dilution method was found to be reversible. These measurements suggest that partial hydrophobic and partial ionic bonding play a role in the interaction of nigerloxin with BSA. PMID- 15134146 TI - Characterization of epoxy carotenoids by fast atom bombardment collision-induced dissociation MS/MS. AB - The characterization and structure of epoxy carotenoids possessing 5,6-epoxy, 5,8 epoxy and 3,6-epoxy end groups conjugated to the polyene chain were investigated using high-energy fast atom bombardment collision-induced dissociation MS/MS methods. In addition to [M - 80](+*), a characteristic fragment ion of an epoxy carotenoid, product ions resulting from the cleavage of C-C bonds in the polyene chain from the epoxy end group, such as m/z 181 (b ion) and 121 (c ion), were detected. On the other hand, diagnostic ions of m/z 286 (e-H ion) and 312 (f-H ion) were observed, not in the 5,6-epoxy or 5,8-epoxy carotenoid but in the 3,6 epoxy carotenoid. These fragmentation patterns can be used to distinguish 3,6 epoxy carotenoids from 5,6-epoxy or 5,8-epoxy carotenoids. The structure of an epoxy carotenoid, 3,6-epoxy-5,6-dihydro-7',8'-didehydro-beta,beta-carotene-5,3' diol (8), isolated from oyster, was characterized using FAB CID-MS/MS by comparing fragmentation patterns with those of related known compounds. PMID- 15134147 TI - Synthesis, isolation, and GC analysis of all the 6,8- to 13,15-cis/trans conjugated linoleic acid isomers. AB - Octadecadienoic acids with conjugated double bonds are often referred to as conjugated linoleic acid, or CLA. CLA is of considerable interest because of potentially beneficial effects reported from animal studies. Analysis of CLA is usually carried out by GC elution of FAME. If the presence of low-level isomers is of interest, a complementary technique such as silverion HPLC is also used. These analyses have been hindered by a lack of well-characterized commercially available reference materials. Described here are the synthesis and isolation of selected 6,8- through 13,15-positional CLA isomers, followed by isomerization of these CLA isomers with iodine to produce all the possible cis,cis, cis,trans, trans,cis, and trans,trans combinations. Also present are the GC retention times of the CLA FAME relative to gamma-linolenic acid (6c,9c,12c-octadecatrienoic acid) FAME using a 100-m CP Sil-88 capillary column (Varian Inc., Lake Forest, CA). These data include all the CLA isomers that have been identified thus far in foods and dietary supplements and should greatly aid in the future analysis of CLA in these products. PMID- 15134148 TI - Levels of N-acylethanolamines in human tumors: in search of reliable data. PMID- 15134149 TI - Sharp-tipped dissecting scissors as an aid in performing endometrial biopsy. AB - OBJECTIVE: To investigate the effectiveness of employing sharp-tipped dissecting scissors as an aid in performing endometrial biopsy (EMB). STUDY DESIGN: Postmenopausal women undergoing routine EMB under approved protocols formed the study group. Three gentle yet firm attempts were made to pass a 3.0-mm Pipelle (Unimar, Wilton, Connecticut) into the uterine cavity. If they were unsuccessful, the closed tips of the dissecting scissors were inserted into the cervical canal and opened. A fourth attempt was then made to pass the Pipelle. RESULTS: A total of 147 postmenopausal women were studied. The average age was 55.7 years, with an average time since menopause of 6.9 years. We were unable to obtain an EMB initially in 11 women (7.5%). After using the scissors, EMB was obtained in 10 of the 11 (90.9%). Age, weight, years since menopause, hormone status and parity had no effect on the ability to perform EMB. Prior cervical surgery was associated with difficult EMB. CONCLUSION: In patients with a tight or closed cervical os, sharp-tipped dissecting scissors can aid in obtaining EMB. PMID- 15134150 TI - Vitamin C and the risk of gestational diabetes mellitus: a case-control study. AB - OBJECTIVE: To examine whether low maternal dietary intake of vitamin C and low maternal plasma ascorbic acid (AA) concentrations are associated with an increased risk of gestational diabetes mellitus (GDM). METHODS: Cases were 67 women with GDM meeting National Diabetes Data Group criteria. Controls were 260 women without such a diagnosis. Maternal dietary vitamin C consumption during the periconceptional period and during pregnancy was assessed using a 121-item, semiquantitative food frequency questionnaire. Maternal plasma AA concentrations were determined using automated enzymatic procedures on specimens collected during the intrapartum period. RESULTS: Mean maternal daily consumption of vitamin C and plasma AA concentrations were 10% and 31% lower, respectively, among GDM cases as compared with controls (130.7 +/- 10.2 vs. 145 +/- 4.9 mg/d, P = .190; 36 +/- 2.0 vs. 53 +/- 1.0 micromol/L, P <.001). After controlling for maternal age, race, prepregnancy adiposity, family history of type 2 diabetes, energy intake and income, women reporting low daily vitamin C intake (< 70 mg/d), as compared with the other women, experienced a 3.7-fold increased risk of GDM (odds ratio [OR] = 3.7, 95% confidence interval [CI] 1.7-8.2). There was a linear relation in risk of GDM with decreasing concentrations of plasma AA (P for linear trend <.001). After adjusting for confounders, women in the lowest quartile (< 42.6 micromol/L), as compared with women in the highest quartile (> 63.3 micromol/L), experienced > 12-fold increased risk of GDM (OR = 12.8, 95% CI 3.5 46.2). CONCLUSION: Low maternal dietary vitamin C intake and low plasma AA concentrations are associated with an increased risk of GDM. Large, prospective, cohort studies are needed to further evaluate the potential beneficial role of vitamin C and other antioxidants in the prevention of impaired glucose tolerance in pregnancy. PMID- 15134151 TI - Summary of safety and effectiveness data from FDA: a valuable source of information on the performance of global endometrial ablation devices. AB - OBJECTIVE: To provide an overview of the approved second-generation endometrial ablation technologies. STUDY DESIGN: Data from the FDA Summary of Safety and Effectiveness Data were compared for Thermachoice, HydroThermablator, Her Option and Novasure devices. RESULTS: At 12 months' follow-up, Novasure and HydroThermablator had the highest amenorrhea rates. Thermochoice and Novasure had the highest success rates at 12 months. Novasure had the lowest adverse event rates in the first 24 hours, between 24 hours and 2 weeks and between 2 weeks and 1 year of follow-up. CONCLUSION: Summary of Safety and Effectiveness Data, obtainable on the FDA Web site, offers objective data for comparing second generation endometrial ablation technologies. PMID- 15134152 TI - Hysteroscopic metroplasty under laparoscopic guidance in infertile women with septate uteri: follow-up of reproductive outcome. AB - OBJECTIVE: To evaluate the effectiveness of hysteroscopic metroplasty under laparoscopic guidance in the treatment of infertile women with a diagnosis of septate uterus and the impact of this surgical procedure on reproductive outcome. STUDY DESIGN: From January 1996 to December 2000, 36 women referred to our endoscopy center underwent hysteroscopic metroplasty. Follow-up covered the incidence of pregnancy, term pregnancies and mode of delivery. RESULTS: The mean age of the patients was 31.9 +/- 3.9 years (range, 23-39). One patient (3%) dropped out during follow-up. There were no intraoperative or postoperative complications. The mean interval from metroplasty to conception was 11.3 +/- 9.2 months (range, 2.0-35.5). Twenty of 35 (57%) women had a history of > or = 1 spontaneous abortions, and 18 of the 20 (90%) achieved pregnancy, with 15 of 18 (83%) term deliveries (10 vaginal deliveries and 5 cesarean sections). Fifteen of 35 patients (43%) were nulliparous, and 8 of 15 (53.3%) had term deliveries (1 cesarean section). CONCLUSION: Hysteroscopic metroplasty under laparoscopic guidance is safe, allows spontaneous delivery and short-term pregnancy planning, is particularly successful in infertile women with a history of > or = 1 spontaneous abortions. PMID- 15134153 TI - Detecting fetal growth restriction or discordant growth in twin gestations stratified by placental chorionicity. AB - OBJECTIVE: To use recommendations made by the Evidence-Based Medicine Working Group to determine the accuracy of sonographically estimated fetal weight to diagnose fetal growth restriction (FGR) (estimated or actual birth weight < 10% for gestational age [GA]) or discordancy in dichorionic and monochiorionic twin gestations. STUDY DESIGN: The inclusion criteria for this retrospective analysis were nonanomalous twins, reliable GA and sonographic estimate of fetal weight within 21 days of live births of both fetuses beyond 26 weeks. For likelihood ratio (LR) and odds ratio (OR), 95% confidence intervals (CIs) were calculated. RESULTS: Over 52 months, 126 twin gestations met the inclusion criteria; 29% were monochorionic and 71% dichorionic. The incidence of FGR in 1 of monochorionic twin fetuses (46%) was twice as high as in dichorionic (26%; OR 2.01; 95% CI 1.00, 4.03), but the rates of discordance > or = 20% were similar (21% vs. 18%, respectively; OR 1.26; 95% CI 0.64, 2.47). The LR (95% CI) of detecting discordant growth > or = 20% was 6.1 (2.5, 15.1) for dichorionic twins and 6.0 (1.8, 20.0) for monochorionic. The LR of identifying FGR in twin A and B in dichorionic twins was 6.3 (2.0, 19.7) and 8.7 (2.9, 26.9), respectively; the corresponding LR for monochorionic twins was 14.5 (1.9, 112.3) and 18.7 (2.6, 135.1). To determine the number of twins required for a tight confidence interval around an LR of 10, post hoc sample size calculations indicated: (1) regardless of chorionicity, attaining LR > 10 is not feasible; (2) in dichorionic twins, detection of FGR with twin A or B is not possible; and (3) about 500 monochorionic pregnancies are necessary to ensure that estimated fetal weight can accurately identify FGR. CONCLUSION: Using guidelines proposed by the Evidence Based Medicine Working Group, at present it may not befeasible to identify aberrant growth in twins. PMID- 15134154 TI - Serum levels of folate, vitamin B12 and homocysteine in complete hydatidiform mole. AB - OBJECTIVE: To investigate the association, if any, between serum concentrations of folate, vitamin B12 and homocysteine in complete hydatiform mole (CHM). STUDY DESIGN: Blood samples were taken from 37 women with CHM and 52 healthy controls in the first trimester of pregnancy. Serum was analyzed for folate, vitamin B12 and homocysteine using electrochemiluminescence and chemiluminescence assay. RESULTS: In patients with CHM, significantly higher mean levels of serum vitamin B12 were encountered (247.21 versus 180.32 pg/mL, P=.012). Significantly lower mean levels of serum folate were found in patients with CHM as compared with controls (7.91 versus 10.72 ng/mL, P=.008). Homocysteine levels tended to be higher in patients with CHM, but the difference was not statistically significant (10.58 versus 8.79 nmol/mL, P=.104). CONCLUSION: This is the first report to suggest an association between folate, vitamin B12 and CHM. Folate may play a protective role in preventing CHM. PMID- 15134155 TI - A nutritional supplement for improving fertility in women: a pilot study. AB - OBJECTIVE: To determine the impact of nutritional supplementation on optimization of reproductive health in women. STUDY DESIGN: A double-blind, placebo-controlled pilot study was initiated to determine the effects of FertilityBlend (Daily Wellness Co., Sunnyvale, California), a proprietary nutritional supplement containing chasteberry and green tea extracts, L-arginine, vitamins (including folate) and minerals. Changes in progesterone level, basal body temperature, menstrual cycle, pregnancy rate and side effects were monitored. RESULTS: Thirty women aged 24-46 years who had tried unsuccessfully to conceive for 6-36 months completed the study. After 3 months, the supplement group (n = 15) demonstrated a trend toward an increase in mean midluteal phase progesterone level (from 8.2 to 12.8 ng/mL, P = .08) and a significant increase in the average number of days in the cycle with basal temperatures >37 degrees C during the luteal phase (6.8-9.7 days, P = .04). The placebo group (n = 15) did not show any notable changes after treatment in any of the parameters studied. After 5 months, 5 of the 15 women in the supplement group were pregnant (33%), and none of the 15 women in the placebo group were (P <.01). No significant side effects were noted. CONCLUSION: Nutritional supplementation may provide an attractive alternative or complement to conventional fertility therapy. PMID- 15134156 TI - Antepartum identification of breech presentation. AB - OBJECTIVE: To determine the accuracy of clinical identification of breech presentation during third-trimester prenatal care and to evaluate factors associated with the accuracy of this determination. STUDY DESIGN: Three hundred eleven patients in the third trimester were prospectively studied. Clinical determination of fetal presentation was made, after which ultrasound was used to determine the true presentation. Amniotic fluid volume, maternal abdominal wall thickness and body mass index were determined for each patient. RESULTS: Among the 311 patients, there were 49 with breech presentation. Twenty-eight of 49 were identified correctly (57.1%). Multiple regression analysis indicated that gestational age, body mass index and maternal abdominal skin thickness did not influence the incidence of correct assessments. There was a significant difference between examiners in the accuracy of clinical assessment of fetal presentation. CONCLUSION: The accuracy of assessment of breech presentation does not appear to depend on gestational age or maternal body mass index but depends more on the individual obstetric care provider. PMID- 15134157 TI - Hemoglobin, altitude and birth weight: does maternal anemia during pregnancy influence fetal growth? AB - OBJECTIVE: To investigate the relationship between maternal hemoglobin concentration, altitude and birth weight. STUDY DESIGN: Birth weights in 235 term pregnancies were investigated for their dependence on maternal hemoglobin concentration after other maternal and pregnancy-specific influences on fetal weight were taken into account. The additional predictive value of hemoglobin concentration on birth weight was assessed using multiple regression. Using published data, the relationship of hemoglobin concentration to altitude was determined, as was the effect of increasing altitude on birth weight. The quantitative effect of hemoglobin concentration on birth weight was correlated with the effect of altitude on hemoglobin concentration to assess whether this could account for the known decrease in birth weight with increasing altitude. RESULTS: Birth weights ranged from 2,220 to 4,850 g (mean, 3,505+/-443), and hemoglobin concentrations ranged from 9.3 to 13.5 g/dL (mean, 11.6+/-0.8). Apart from other known predictive variables, the variation in maternal hemoglobin concentrations at constant altitude independently explained 2.6% of the variance in birth weight (r=-.18, P=.003). Term birth weight was reduced by 89 g for each 1.0 g/dL increase in hemoglobin concentration (P<.01). For every 1,000-m increase in altitude, hemoglobin concentration increased by 1.52 g/dL and birth weight decreased by 117 g. CONCLUSION: Birth weight correlates negatively with maternal hemoglobin concentration. This is consistent with the well-known effect of high altitude exposure during pregnancy, which increases both hematocrit and blood viscosity and lowers birth weight. The quantitative effect on birth weight of increasing maternal hemoglobin concentration at constant altitude is within 13% of the change in birth weight that can be attributed to the change in hemoglobin concentration associated with increases in altitude. PMID- 15134158 TI - Perineal body length and lacerations at delivery. AB - OBJECTIVE: To define normal perineal body length during labor and determine if a shortened perineal body is associated with perineal lacerations or operative vaginal delivery. STUDY DESIGN: We reviewed charts of patients admitted for labor over a 4-month period. The perineal body was measured by the admitting physician and delivery outcomes obtained from inpatient records. Patients were excluded for malpresentation, multiple gestation, gestational age < 36 weeks, incomplete records and scheduled cesarean delivery. To determine if differences existed between patients with perineal body measurements available and those without, chi2 analysis was used, with P<.05 considered significant. Multiple logistic regression was used to control for confounding variables and determine if a shortened perineal body affected the incidence of operative vaginal delivery and significant lacerations at vaginal delivery. RESULTS: A total of 234 patients met our inclusion criteria; perineal body measurements were available for 133 (57%). The average perineal body length was 3.90 cm (+/-0.70). Patients with a perineal body of < or = 2.5 cm had a significantly higher chance of sustaining a third- or fourth-degree laceration (40% vs. 5.6%, P=.004). This risk remained after controlling for both operative vaginal delivery and episiotomy. The incidence of operative vaginal delivery was greater (28.5% vs. 9.2%, P =.006) for patients with a perineal body < or = 3.5 cm. CONCLUSION: There is an increased risk of significant lacerations and operative vaginal delivery in patients with a shortened perineal body. PMID- 15134159 TI - Considerations in the choice of oral vs. transdermal hormone therapy: a review. AB - The benefit/risk profile of postmenopausal hormone therapy has been under greater scrutiny since Women's Health Initiative study data were published in July 2002. A nominal analysis showed the study drug, a combination of 0.625 mg conjugated equine estrogens (CEE) and 2.5 mg medroxyprogesterone acetate (MPA), to be associated with decreased risks of colorectal cancer and osteoporotic hip fractures but increased risks of coronary heart disease, stroke and venous thromboembolic events. This same profile was not seen in the estrogen-only arm of the study, and the trial investigators cautioned that the results observed with CEE/MPA might not apply to other dosages of combined hormone therapy, other types of estrogens and progestins, or other routes of administration. However, the unexpected results cast a long shadow of doubt over the safety of all hormone therapy, making it difficult for reproductive health care professionals to counsel women on the selection of appropriate treatment of vasomotor symptoms. A review of the literature suggests that not all estrogens and progestins are alike, and alternative drugs, doses and delivery systems may exhibit better safety profiles than CEE/MPA, with no loss of efficacy. Selection of therapy should be individualized, based on patients' specific needs and global health risks. When there is a compelling need for relief of climacteric symptoms, using the lowest effective dose of hormonal therapy remains a prudent recommendation. PMID- 15134160 TI - Endometriosis presenting as a urethral diverticulum: a case report. AB - BACKGROUND: Pelvic pain is a common complaint among women of childbearing age. It has an extensive differential diagnosis that at times can make it difficult to determine its etiology. One must therefore rely on the characteristics of the physical examination, symptoms and imaging studies. However, in doing so, one should keep in mind that many diseases mimic one another. Physicians must be careful not to fall into the trap of simply assigning a specific disease to a given group of symptoms. CASE: A 35-year-old woman, gravida 2, para 0020, presented to a clinic complaining of left lower abdominal pain. She had a history of dyspareunia, dysmenorrhea, urinary frequency and numerous urinary tract infections. Previous laparoscopies had been negative for endometriosis. Physical examination demonstrated a 1.5-cm mass left of the midurethra. No pus was expressed through the urethra with cyst massage. Imaging showed a 1.1 x 1.1-cm lesion in the left posterolateral aspect of the urethra consistent with a urethral diverticulum. Uterine adenomyosis was also noted. Although clinical symptoms, physical examination and imaging suggested a urethral diverticulum, a vaginal endometriotic cyst was encountered at surgery. Pathologic evaluation of the surgically excised lesion revealed endometriosis, revealed endometriosis. CONCLUSION: In this case, clinical findings, location and imaging characteristics of a periurethral endometriotic lesion suggested a urethral diverticulum. Endometriosis should be considered in patients with a history of pelvic pain who present with urinary frequency and a periurethral lesion. PMID- 15134161 TI - Low grade uterine stromal sarcoma and squamous cell carcinoma associated with an IUD: a report of 2 cases. AB - BACKGROUND: Although intrauterine device (IUD) usage has not been linked to uterine cancer and has even been considered protective against neoplasia, sporadic cases of IUD-associated uterine malignancies have been reported. CASES: One cystic low grade stromal sarcoma of the uterus occurred in a 7-year IUD user, and 1 invasive endometrial squamous cell carcinoma following 25 years of IUD retention occurred in another. The latter case was notable for extensive in situ squamous cell carcinoma involving the entire uterine cavity and extending to the ectocervix and into both fallopian tubes. In situ hybridization for high-risk HPV DNA was negative. These findings raise the possibility of a non-HPV-related pathway for the squamous cell carcinoma case due possibly to prolonged chronic irritation and pyometra. CONCLUSION: The association of low grade stromal sarcoma with the IUD in the first case is extremely rare and most likely coincidental. The squamous carcinoma case, however, may have arisen via a non-HPV-related pathway, possibly related to IUD-induced chronic irritation and pyometra. PMID- 15134162 TI - MEPs vote to amend transport regulation. PMID- 15134163 TI - Perioperative deaths in small animals: findings so far. PMID- 15134164 TI - Dietary trials with a commercial chicken hydrolysate diet in 63 pruritic dogs. AB - The owners of 63 pruritic dogs were instructed to feed them a chicken hydrolysate diet exclusively for six weeks as part of diagnostic investigations into non seasonal pruritus. Ectoparasitism and microbial infections were eliminated during the dietary trial. The dogs' skin lesions, gastrointestinal signs and frequency of defecation were assessed and scores for pruritus were assigned before they started the diet and before and after they resumed their original diet. An adverse food reaction was diagnosed if the pruritus resolved while they were on the diet, but recurred when they resumed their original food regimen. Seventeen of the 63 dogs were withdrawn from the trial, including four which found the diet unpalatable; however, its palatability was reported to be good or excellent in 48 of the dogs. An adverse food reaction alone was diagnosed in nine (19.6 per cent) of the 46 dogs and another nine had an adverse food reaction and atopy. There were gastrointestinal signs in six of the nine dogs with an adverse food reaction, which resolved on the trial diet but recurred when they resumed their original diet. PMID- 15134165 TI - Distemper virus as a cause of central nervous disease and death in badgers (Meles meles) in Denmark. AB - During the summer of 2002 a distemper-like disease was observed in the free ranging badger population in Denmark. It was characterised by grand seizures, abnormal behaviour and death; the badgers all had severe chronic pneumonia and some had non-suppurative encephalomyelitis. In this study, eight of the affected badgers were examined by gross pathological, histological, immunohistological, bacteriological, parasitological and virological methods, and were diagnosed with distemper; canine distemper virus was identified. PMID- 15134166 TI - Compensatory humeral overgrowth associated with antebrachial shortening in six dogs. PMID- 15134167 TI - Cerebral theileriosis in a Holstein calf. PMID- 15134168 TI - Osseous cyst-like lesion of the intermediate humeral tubercle of a horse. PMID- 15134169 TI - Treatment of sterile panniculitis in a dog. PMID- 15134170 TI - Correct use of intramammary treatments at drying off. PMID- 15134172 TI - Mortality in budgerigars in Scotland: pathological findings. PMID- 15134171 TI - Funding animal health and welfare. PMID- 15134173 TI - Mortality in budgerigars associated with a reovirus-like agent. PMID- 15134175 TI - Donations to the VBF. PMID- 15134174 TI - Buffalo rabies in Nepal. PMID- 15134176 TI - Lamivudine-resistant hepatitis B virus and ongoing lamivudine therapy: stop the merry-go-round, it's time to get off! PMID- 15134177 TI - Comparisons of the HBV and HIV polymerase, and antiviral resistance mutations. AB - The antiviral treatment of chronic hepatitis B is limited by the selection of antiviral resistance mutations. Primary resistance to lamivudine occurs at rtM2041/V in the C Domain of the polymerase. Recently, resistance to adefovir has also been described in the D Domain at rtN236T. The treatment of patients with resistant virus without complete suppression can lead to the further selection of compensatory mutations. Thus, to gain an understanding of the hepatitis B virus (HBV) polymerase and also mutations associated with resistance, a three dimensional model of the HBV reverse transcriptase core region based on homology with human immunodeficiency virus (HIV) was created. A comparative analysis of the HIV polymerase and the model of HBV polymerase was performed. In addition, the antiviral resistance mutations including potential compensatory mutations were mapped to determine their effect on the HBV polymerase model, especially in the nucleotide binding site. PMID- 15134178 TI - Adipocytes targets and actors in the pathogenesis of HIV-associated lipodystrophy and metabolic alterations. AB - The recent clinical use of potent HIV-1 drugs, including nucleoside reverse transcriptase inhibitors (NRTIs) and non-peptidic viral protease inhibitors (PIs), and their combinations, termed highly active antiretroviral therapy (HAART), has dramatically reduced the infection-related mortality of AIDS patients, but it is associated with severe metabolic adverse events such as lipodystrophy syndrome, dyslipidaemia, insulin resistance and diabetes mellitus. The aetiology of this syndrome and metabolic alterations appear to be multifactorial, including HIV drug inhibitory effects on adipocyte differentiation, alteration of mitochondrial functions in adipocytes and altered leptin, adiponectin and cytokine expression in adipose tissue of patients. Adipose tissue may thus be a central regulator in disorganized lipid metabolism and insulin resistance associated with antiretroviral therapy, and we propose in this review to explore how adipose tissue may be a target, but also an actor, in the aetiopathogenesis of the lipodystrophy syndrome. PMID- 15134180 TI - Skin reactions in patients with influenza treated with oseltamivir: a retrospective cohort study. AB - Oseltamivir phosphate is an FDA-approved treatment for influenza that has been available for prescription use in the USA since 1999. The present report describes findings from a post-marketing safety study of skin reactions associated with oseltamivir use. All patients in the claims-derived Ingenix Research Database with a physician diagnosis of influenza and/or a dispensing of oseltamivir between 1 December 1999 and 31 March 2002 were identified. Cohort eligibility criteria included minimum baseline enrolment duration of 3 months, age of at least 1 year and no influenza vaccination on the date of influenza diagnosis or oseltamivir dispensing. Patients were classified into two primary cohorts, influenza diagnosis and oseltamivir dispensing on the same day, and influenza diagnosis but no oseltamivir at any time, and a cohort included for secondary analyses comprising patients who received an oseltamivir dispensing without an influenza diagnosis on the same day. Outcomes included general skin reactions and several specific skin reactions. Events occurring during the 30 days following the date of influenza diagnosis or oseltamivir dispensing were examined using Cox proportional hazards models. Model covariates included age, use of another influenza drug, month and year of index date, and use of antitussives. Adjusted rate ratios for the general class of skin reactions among the primary cohort of oseltamivir users versus non-users were 1.05 (95% CI: 0.88 1.24) for incident cases and 0.98 (95% CI: 0.77-1.24) among patients with a history of a skin reaction. Similar results were seen for the other skin reaction categories, and secondary analyses investigating the oseltamivir users without influenza revealed no elevation in risk. It is concluded that oseltamivir use does not appear to be associated with an increased risk of skin reactions. PMID- 15134179 TI - Influence of mother and infant zidovudine treatment duration on the age at which HIV infection can be detected by polymerase chain reaction in infants. AB - OBJECTIVE: To investigate the influence of zidovudine (ZDV) prophylaxis duration in mothers and infants on the age at which infection becomes detectable by DNA polymerase chain reaction (PCR) in non-breastfed infants. METHODS: Blood samples were collected sequentially from birth to 6 months in a Thailand perinatal HIV prevention trial in which 98 transmissions occurred. The proportions of infections detectable at birth and the Turnbull distributions of age at which infection became detectable after birth were compared according to actual ZDV treatment duration (mothers: no more than 7.5 weeks versus more; infants: 3 days versus at least 4 weeks), provided an adherence greater than 75%. RESULTS: Detectable infection at birth was less frequent in children whose mothers received a long treatment as compared to a short treatment (27 vs 50%, P=0.04). When mothers received a long treatment, infant ZDV treatment duration did not influence the distribution of age at which infection became detectable after birth (median 24 days). However, when mothers received a short treatment, this distribution was shifted to the right when infants received a long treatment (median 43 days, P<0.0001), and to the left when infants received a short treatment (median 11 days, P<0.0001). CONCLUSIONS: When mothers receive a short treatment, the proportion of infections detectable at birth is higher and the time at which infection becomes detectable after birth depends on the infant treatment duration. In the study conditions, a PCR result after 2 months could be used to define infection status. PMID- 15134182 TI - Asymptomatic hyperlactataemia: predictive value, natural history and correlates. AB - BACKGROUND: The significance of asymptomatic hyperlactataemia is unknown. METHODS: We measured serial lactate levels in a group of HIV-infected subjects. Objectives were to identify covariates associated with hyperlactataemia, and to explore the natural history of hyperlactataemia. RESULTS: Overall, 1487 lactate measurements were performed on 396 subjects; 299 subjects had serial lactate testing, with a median of four performed per patient (range: 2-16). At study entry, lactate was >2.4 mmol/l in 6% of subjects and >3.6 mmol/l in 1%. A multiple logistic regression analysis revealed that cholesterol and current stavudine were the only predictors of hyperlactataemia. Lactate levels were highly reproducible both for short-term (within 3 months) and long-term (>1 year) follow-up. During the study period, 16 subjects were identified with sustained hyperlactataemia; 12/16 remained asymptomatic after a median of 210 days (30 585), and four developed symptoms suggestive of lactic acidosis syndrome. In three of the four, asymptomatic hyperlactataemia had preceded the onset of symptoms by 18-122 days. A longitudinal model showed a rapid rise of lactate levels from 0 to 12 months on nucleoside reverse transcriptase inhibitor (NRTI) and then stabilization. Lactate levels did not correlate with baseline or on study development of lipoatrophy. CONCLUSIONS: Even when optimal methods of collection are used, asymptomatic hyperlactataemia does exist and could be sustained in a subset of NRTI-treated subjects for as long as 585 days. A subset of these subjects became symptomatic, after as long as 122 days of asymptomatic hyperlactataemia. Asymptomatic hyperlactataemia is not predictive of lipoatrophy. PMID- 15134181 TI - Immune restoration in HIV-positive, antiretroviral-naive patients after 1 year of zidovudine/lamivudine plus nelfinavir or nevirapine. AB - OBJECTIVES: To evaluate the immunological response in HIV-1-infected, antiretroviral-naive patients receiving highly active antiretroviral therapy regimen of two nucleosides plus a protease inhibitor or a non-nucleoside reverse transcriptase inhibitor. DESIGN AND METHODS: Of 142 patients included in a randomized, open, multicentre trial comparing zidovudine/lamivudine plus nelfinavir (NFV) or nevirapine (NVP), 36 patients (16 NFV, 20 NVP) were enrolled in an immunological substudy. Mean baseline CD4 T-cell counts was 360/mm3 (range: 11-679) and mean baseline plasma viral load >50000 copies/ml (range: 2240 1468210). Viral load (VL), T-cell subsets and T-cell functions were analysed at baseline and after 1 year of treatment. RESULTS: After 12 months of follow-up, plasma viral load was reduced similarly in both groups, with 78% (NFV) and 83% (NVP) of patients achieving a VL <200 copies/ml. A significant increase in CD4 T cells was observed in both groups (mean: +182 cells, P=0.001). Both regimens were similarly effective in reducing activated T cells (CD38 and DR). A significant increase of both CD4 and CD8 CD28 T cells occurred in both arms of treatment. Patients of both regimens showed a significant decrease of activated memory (CD45RA-CD45RO+) CD8 T cells and a clear increase of naive (CD45RA+CD45RO-) CD8 T cells. Peripheral blood mononuclear cell proliferative responses to polyclonal stimuli (CD3 and CD3 +CD28) as well as to ubiquitous cytomegalovirus antigen increased significantly in both groups after 12 months of follow-up. Nevertheless, neither at baseline nor after 1 year of treatment, these patients showed any significant T-cell responsiveness to HIV-1 recombinant proteins gp160 or p24. CONCLUSIONS: Our data indicate that immune restoration achieved after 1 year of therapy with either NFV or NVP was similar. This reinforces the role of NVP-containing regimens as a valid option for initiating antiretroviral therapy. Nevertheless, additional therapeutic approaches should be envisaged to restore HIV-1-specific T-cell responses. PMID- 15134183 TI - Comparison of two reduced-dose regimens of indinavir (600 mg vs 400 mg twice daily) and ritonavir (100 mg twice daily) in healthy volunteers (COREDIR). AB - OBJECTIVE: To assess the pharmacokinetics and tolerability of reduced dosages of twice daily indinavir (IDV) boosted by low-dose ritonavir (RTV) in healthy volunteers. METHODS: Pharmacokinetics and tolerability of IDV/RTV twice daily (600/100 mg and 400/100 mg) were assessed in a randomized crossover design in 16 healthy volunteers. Each dosage was taken twice daily for 2 weeks before 12 h pharmacokinetics were obtained. RESULTS: Sixteen subjects were included, with a mean age +/- SD of 30 +/- 4 years; seven female, nine male. Fifteen subjects completed the study. After dose reduction of IDV AUC, Cmax and Cmin decreased significantly. In the 400 mg group three out of 15 subjects had IDV levels below 0.10 mg/l vs none in the 600 mg group. All subjects reported mild to moderate side effects throughout the study period, which were more severe in the 600 mg group (mostly renal, dry skin/lips, paresthesias/oral discomfort). In the 600 mg group four subjects reported dysuria and one subject discontinued because of flank pain, whereas two subjects reported dysuria and no subject discontinued in the 400 mg group, respectively. Eight subjects developed crystalluria without a significant difference between both groups. No significant change in serum creatinine was observed. CONCLUSIONS: IDV/RTV 400/100 mg twice daily resulted in significant lower IDV exposure, with three out of 15 subjects revealing Cmin values below the recommended threshold for wild-type virus of 0.10 mg/l. Tolerability, however, was lower in the 600 mg IDV group. Therapeutic drug monitoring in the individual patient appears to be necessary to guarantee appropriate drug levels and simultaneously minimize toxicity. PMID- 15134184 TI - Impact of insertions in the HIV-1 p6 PTAPP region on the virological response to amprenavir. AB - We evaluated the impact of genetic changes within p6Gag gene on the virological response (VR, mean decrease in plasma viral load at week 12) to unboosted amprenavir (APV). Gag-protease fragments, including gag p2, p7, p1, p6 regions and whole protease (PR) were sequenced from baseline plasma specimens of 84 highly pre-treated but APV-naive patients included in the NARVAL (ANRS 088) trial. The correlation between baseline p6Gag polymorphism, PR mutations, baseline characteristics and VR to APV was analysed in univariate analysis. Insertions (P459Ins) within p6 protein, leading to partial or complete duplication of the PTAPP motif, were significantly associated with a decreased VR (P459Ins versus wild-type; -0.3 +/- 0.8 vs -1.1 +/- 1.2 log copies/ml, P=0.007) and were more frequent when the V82A/F/T/S PR mutation was present (P=0.020). In multivariate analysis, after adjustment on the predictive factors of the VR in the NARVAL trial and on the PR mutations linked with response, there was a strong trend to an association (P=0.058) between the presence of P459Ins and an altered VR. In conclusion, these results suggest that insertions in the p6 region of HIV 1 gag gene may affect the VR, in highly pre-treated patients receiving an unboosted APV-containing regimen. PMID- 15134185 TI - Adherence to antiretroviral therapy and CD4 T-cell count responses among HIV infected injection drug users. AB - OBJECTIVE: To evaluate the time to CD4 cell count response (> or = 50 cells/mm3) among patients initiating highly active antiretroviral therapy (HAART) with and without a history of injection drug use, and to examine the potential role of non adherence to HAART on differential CD4 responses. METHODS: Population-based analysis of treatment-naive patients initiating HAART during the period 1 August 1996 to 31 July 2000 and who were followed until 31 March 2002. Patients were stratified based on 95% adherence and history of injection drug use, and Kaplan Meier methods and Cox regression were used to evaluate CD4 response rates and factors associated with CD4 responses. RESULTS: Overall, the CD4 cell count response rate was slower among injection drug users in Kaplan-Meier analyses (log rank: P<0.05). However, no differences existed when the analyses were restricted to adherent patients (log-rank: P=0.349). Similarly, the differences in the time to CD4 cell count response observed in univariate Cox regression analyses for patients with a history of injection drug use [relative hazard: 0.85 (95% CI: 0.75-0.97)] diminished after adjustment for adherence [adjusted relative hazard: 1.02 (95% CI: 0.89-1.16)]. CONCLUSION: These data demonstrate the importance of adherence on CD4 cell count responses and highlight the need for interventions to improve antiretroviral adherence among injection drug user. PMID- 15134186 TI - Estimates of intracellular delay and average drug efficacy from viral load data of HIV-infected individuals under antiretroviral therapy. AB - We analyse viral load data of five patients under ritonavir monotherapy using a model of HIV dynamics under antiretroviral therapy that includes both drug pharmacokinetics and the intracellular delay from the time of cell infection to viral production. Using this approach we separate pharamacokinetic from intracellular delays, and obtain new estimates of intracellular delay and the antiviral efficacy of ritonavir. We find the average intracellular delay to be 1 day, in agreement with experiments. The average viral generation time is now estimated at 2 days, resulting in approximately 180 replication cycles per year. The model also reveals that ritonavir monotherapy is approximately 65% as efficacious as a recently used potent four-drug therapy, suggesting that selection for drug resistance may be facilitated by the relatively low efficacy of individual drugs, contributing in part to the inherent limitations of current therapies in combating HIV-1 infection. PMID- 15134187 TI - Once-daily dosing of saquinavir soft-gel capsules and ritonavir combination in HIV-1-infected patients (IMEA015 study). AB - This was a prospective pilot study evaluating a saquinavir (SQV) soft-gel capsules (SGC)/ritonavir (RTV)-containing once-daily regimen over a follow-up of 3 months. The primary end-point was to determine the number of patients both remaining on treatment at month 3 and with trough SQV plasma concentration 24 h after the last intake (C24h) exceeding the inhibition of 95% of viral replication in vitro (IC95). The secondary end-points were to investigate the immuno virological efficacy and safety of SQV-SGC/RTV once daily, and to explore SQV concentrations in peripheral blood mononuclear cells (PBMCs). Twenty-three antiretroviral-naive and 17 protease inhibitors (PIs) experienced HIV-1-infected patients with plasma HIV-1 RNA level below 200 copies/ml were enrolled. They were assigned to SQV-SGC/RTV (1600/100 mg once daily) combined with nucleoside and/or non-nucleoside reverse transcriptase inhibitors. In a subgroup of 13 patients, both plasma and intracellular SQV concentrations were determined. By intent to treat analysis the percentage of success at month 3 was 87.5% (confidence interval: 73.2-95.8%) with 78.3% in naive and 100% in PI-experienced patients. SQV C24h and intracellular concentrations [median (range, n)] were 241 ng/ml (40 1209, 35) and 323 ng/ml (168-475, 12), respectively. Intracellular concentrations showed an accumulation of SQV in PBMCs persisting during 24 h. Neither immunological nor virological failure was observed. Clinical and biological tolerance was acceptable in all patients but three with adverse effects leading to discontinuation. These data confirmed the short-term efficacy of SQV-SGC/RTV once-daily regimen based on SQV therapeutic drug monitoring. PMID- 15134188 TI - No benefit to continue lamivudine therapy after emergence of YMDD mutations. AB - BACKGROUND: Mutations in the sequence of the conserved tyrosine-methionine aspartate-aspartate (YMDD) locus of the HBV RNA-dependent DNA polymerase (rt M 204 I/V) may develop after 6-9 months of lamivudine therapy. The current practice of continuing lamivudine therapy has been associated with hepatitis flares or even hepatic decompensation. In addition, experiments have shown that the defective replication competency of rt M 204 I/V restores upon addition of lamivudine. AIM: To evaluate whether continuing lamivudine therapy after emergence of rt M 204 I/V is appropriate. PATIENTS AND METHODS: The clinical courses of 66 patients with continuing lamivudine therapy (continued group) and 68 patients who discontinued lamivudine therapy (discontinued group) were monitored monthly or more frequently if condition required. Hepatitis flare, jaundice, hepatic decompensation and hepatitis B e antigen (HBeAg) seroconversion in HBeAg-positive patients were documented as events. RESULTS: In a 12-month period, hepatitis flares and decompensations occurred in 67 and 11%, respectively, in patients who continued lamivudine therapy, as compared with 54 and 7%, respectively (P>0.05), in those who stopped therapy. HBeAg seroconversion rate was 19% in continued group and 35% in discontinued group (P=0.08). Serum HBV DNA increased in 48 (73%) of the continued group, and the median level increased from 46 pg/ml upon first detection of mutation to 330 pg/ml (P<0.001) at the end of 12 months continuing therapy. In contrast, serum HBV DNA level in the discontinued group increased in 22 (33%) patients but decreased in 39 patients, and median level decreased from 172 to 55 pg/ml at the end of 12 months after stopping lamivudine. CONCLUSION: These results suggest that there is no benefit to continued lamivudine therapy after emergence of rt M 204 I/V. PMID- 15134189 TI - Long-term virological response to multiple sequential regimens of highly active antiretroviral therapy for HIV infection. AB - OBJECTIVE: Information about the virological response to sequential highly active antiretroviral therapy (HAART) for HIV infection is limited. The virological response to four consecutive therapies was evaluated in the Swiss HIV Cohort. DESIGN: Retrospective analysis in an observational cohort. METHODS: 1140 individuals receiving uninterrupted HAART for 4.8 +/- 0.6 years were included. The virological response was classified as success (<400 copies/ml), low-level (LF: 400-5000 copies/ml) or high-level failure (HF: >5000 copies/ml). Potential determinants of the virological response, including patient demographics, treatment history and virological response to previous HAART regimens were analysed using survival and logistic regression analyses. RESULTS: 40.1% failed virologically on the first (22.0% LF; 18.1% HF), 35.1% on the second (14.2% LF; 20.9% HF), 34.2% on the third (9.9% LF; 24.3% HF) and 32.7% on the fourth HAART regimen (9% LF; 23.7% HF). Nucleoside pre-treatment (OR: 2.34; 95% CI: 1.67-3.29) and low baseline CD4 T-cell count (OR: 0.79/100 cells rise; 95% CI: 0.72-0.88) increased the risk of HF on the first HAART. Virological failure on HAART with HIV-1 RNA levels exceeding 1000 copies/ml predicted a poor virological response to subsequent HAART regimens. A switch from a protease inhibitor- to a non nucleoside reverse transcriptase inhibitor-containing regimen significantly reduced the risk of HF. Multiple switches of HAART did not affect the recovery of CD4 T lymphocytes. CONCLUSION: Multiple sequential HAART regimens do not per se reduce the likelihood of long-term virological suppression and immunological recovery. However, early virological failure increases significantly the risk of subsequent unfavourable virological responses. The choice of a potent initial antiretroviral drug regimen is therefore critical. PMID- 15134190 TI - Meta-analysis of mutations in the NS5A gene and hepatitis C virus resistance to interferon therapy: uniting discordant conclusions. AB - BACKGROUND: Hepatitis C virus genotype 1B responds poorly to treatment with interferon, in contrast to the more interferon-sensitive genotypes 2 and 3. Studies on combination therapy regimens with PEG-interferon and ribavirin report sustained response rates that generally do not exceed 50%, in contrast to sustained response rates of 80% for genotype 2 and 3. In Japan, a correlation was found between the number of mutations in an 'interferon sensitivity determining region' (ISDR) and outcome of interferon treatment in genotype 1B-infected patients. However, an ongoing controversy on the existence of an ISDR in non Japanese isolates resulted, as non-Japanese studies failed to confirm this association. The present study approached this issue by carrying out a meta analysis of ISDR sequences and response to interferon treatment. METHODS: Twenty seven studies were included, reporting 1351 ISDR sequence data of genotype 1B infected patients and their virological response to interferon treatment. Both summary statistics and individual patient data were used systematically to explore the association between ISDR mutations and response to interferon. RESULTS: The ISDR effect on response was universally present but appeared to be stronger in Japan, with a relative risk of 5.73 for mutant viruses as compared to 4.66 for non-Japanese isolates. High interferon dose, in Japan administered more frequently, was associated with an increase in response rate only among patients infected with mutant isolates. Interaction between dose and ISDR type was confirmed in a logistic regression model. After stratifying for dose, differences in response rate between Japanese and non-Japanese patients were no longer present. CONCLUSION: This study puts an end to a longstanding controversy by confirming the universal existence of an ISDR in genotype 1B-infected patients. Apparent discrepant findings from Japanese and non-Japanese studies can be explained by differences in dosing regimens and a dose-dependent differential effect of ISDR mutations on response to treatment. PMID- 15134191 TI - Probing the structure of the SARS coronavirus using scanning electron microscopy. AB - A novel coronavirus, SARS-CoV, has been confirmed to be the aetiological agent of SARS. Transmission electron microscope (TEM) images played an important role in initial identification of the pathogen. In order to obtain greater morphological detail of SARS-CoV than could be obtained by TEM, we used ultra-high resolution scanning electron microscopy (SEM) to image the virus particles. We show here the three-dimensional appearance of SARS-CoV. Enhanced detail of the ultrastructure reveals the trimeric structure of the 10-20 nm spikes on the virion surface. These results contribute to characterization of the SARS agent and development of new antiviral strategies. PMID- 15134192 TI - Relative prognostic value of self-reported adherence and plasma NNRTI/PI concentrations to predict virological rebound in patients initially responding to HAART. AB - We studied the predictive value of self-reported adherence and plasma drug concentrations on virological rebound to HAART. Among 238 participants in the AdICoNA study who had viral load < or = 500 copies/ml, 42 (17.6%) experienced virological rebound by 96 weeks. Both self-reported non-adherence and sub-optimal concentration were independently associated with a higher risk of virological rebound. PMID- 15134194 TI - Beetling around the genome. AB - The red flour beetle, Tribolium castaneum, has been selected for whole genome shotgun sequencing in the next year. In this minireview, we discuss some of the genetic and genomic tools and biological properties of Tribolium that have established its importance as an organism for agricultural and biomedical research as well as for studies of development and evolution. A Tribolium genomic database, Beetlebase, is being constructed to integrate genetic, genomic and biological data as it becomes available. PMID- 15134195 TI - Phenotype of the Triplo-lethal locus of Drosophila melanogaster and its suppression by hyperoxia. AB - The Triplo-lethal locus (Tpl) of Drosophila is both triplo-lethal and haploinsufficient, but the function of the locus is unknown. We have examined Tpl aneuploid embryos and find that, in both trisomics and monosomics, the midgut shows extensive cell death and the tracheae are abnormal. Shortly thereafter, all tissues die. PCR-based genotyping of individual embryos and larvae show that this phenotype occurs in the trisomics after hatching and in the monosomics before hatching. Weak alleles of the interacting gene Su(Tpl) delay the death of Tpl trisomics, but they still show the same tracheal and midgut phenotypes before dying. Hyperoxia (45% oxygen) partially suppresses the phenotype of Tpl aneuploids, even though the use of a hypoxia reporter strain shows that dying Tpl aneuploids are not hypoxic. This is the first report of a phenotype associated with the Tpl locus and the first report of an environmental condition that suppresses the phenotype. PMID- 15134193 TI - Safety and efficacy of a NRTI-sparing HAART regimen of efavirenz and lopinavir/ritonavir in HIV-1-infected children. AB - We studied a nucleoside reverse transcriptase inhibitor (NRTI)-sparing regimen for the treatment of children infected with NRTI-resistant HIV-1. The combination of lopinavir/ritonavir and efavirenz suppressed HIV-1 levels for a prolonged period and resulted in a significant increase in CD4+ T cell numbers despite an extensive prior treatment with NRTI (>4 years). Observed side effects were transient with the exception of dyslipidaemia. PMID- 15134196 TI - Variation over space and time of Aedes aegypti in Phnom Penh (Cambodia): genetic structure and oral susceptibility to a dengue virus. AB - We studied spatial and temporal variation in 20-23 Aedes aegypti samples collected in Phnom Penh and its suburbs to estimate the population genetic structure using allozymes and the susceptibility to a dengue-2 virus. Based on seven allozyme systems, we detected low levels of genetic exchanges (i.e. high, significant F(ST) values) between populations collected in the city centre, and different patterns of genetic structure for samples collected in the suburbs, depending on the type of environment and the date of collection. In the southern suburbs and the Chroy Chang Var Peninsula, differentiation became highly significant at the end of the dry season, whereas the opposite situation was observed for collections from the northern suburbs. Vector competence assessed by oral infections with a dengue-2 virus was lower for samples collected in the city centre than in the suburbs. A significant decrease of dengue susceptibility was observed in populations during the dry season. This study allows a model of Ae. aegypti population functioning in Phnom Penh to be suggested. Dynamics of dengue virus diffusion depend on the population genetic structure of the vector and its evolution over space and time. PMID- 15134197 TI - Coalescence times and the Meselson effect in asexual eukaryotes. AB - In asexual eukaryotes, the two allelic gene copies at a locus are expected to become highly divergent as a result of the independent accumulation of mutations in the absence of segregation. If sexual reproduction was abandoned millions of generations ago, intra-individual allelic divergences can be significantly larger than in species that reproduce sexually. Owing to the disputed existence of truly ancient asexual species, this so-called 'Meselson effect' has been put forward as a means of confirming the complete loss of sexual reproduction. Very few attempts have, however, been made at quantifying the effect of sexual reproduction on the degree of divergence between gene copies in an asexual population. Here, I describe how asexual reproduction can be regarded as a special case of population subdivision. Using a slightly modified version of the standard two-deme structured coalescent, I derive the expected coalescence time for a pair of gene copies in an asexual population and show that the Meselson effect is compatible with low rates of sexual reproduction. PMID- 15134198 TI - Estimating numbers of EMS-induced mutations affecting life history traits in Caenorhabditis elegans in crosses between inbred sublines. AB - Inbred lines of the nematode Caenorhabditis elegans containing independent EMS induced mutations were crossed to the ancestral wild-type strain (N2). Replicated inbred sublines were generated from the F1 offspring under conditions of minimal selection and, along with the N2 and mutant progenitor lines, were assayed for several fitness correlates including relative fitness (w). A modification of the Castle-Wright estimator and a maximum-likelihood (ML) method were used to estimate the numbers and effects of detectable mutations affecting these characters. The ML method allows for variation in mutational effects by fitting either one or two classes of mutational effect, and uses a Box-Cox power transformation of residual values to account for a skewed distribution of residuals. Both the Castle-Wright and the ML analyses suggest that most of the variation among sublines was due to a few (approximately 1.5-2.5 on average) large-effect mutations. Under ML, a model with two classes of mutational effects, including a class with small effects, fitted better than a single mutation class model, although not significantly better. Nonetheless, given that we expect there to be many mutations induced per line, our results support the hypothesis that mutations vary widely in their effects. PMID- 15134199 TI - Normal linear models with genetically structured residual variance heterogeneity: a case study. AB - Normal mixed models with different levels of heterogeneity in the residual variance are fitted to pig litter size data. Exploratory analysis and model assessment is based on examination of various posterior predictive distributions. Comparisons based on Bayes factors and related criteria favour models with a genetically structured residual variance heterogeneity. There is, moreover, strong evidence of a negative correlation between the additive genetic values affecting litter size and those affecting residual variance. The models are also compared according to the purposes for which they might be used, such as prediction of 'future' data, inference about response to selection and ranking candidates for selection. A brief discussion is given of some implications for selection of the genetically structured residual variance model. PMID- 15134200 TI - Economic burden of depression in Pakistan. PMID- 15134201 TI - Prevalence and factors associated with anxiety and depression among family practitioners in Karachi, Pakistan. AB - OBJECTIVE: We aimed to assess anxiety and depression and to identify the factors associated with these conditions among family practitioners in Karachi, Pakistan. METHODS: A self-reported postal survey was conducted among 540 family practitioners in Karachi during the months of July - August 2003. Aga Khan University Anxiety and Depression Scale (AKUADS) was used to assess anxiety and depression. Additional questions were also inquired to get information about socio-demographic, professional and lifestyle characters. RESULTS: Using AKUADS, 155 (39%) family practitioners had anxiety and depression. Mutlivariate analysis disclosed five factors to be significantly associated with anxiety and depression; these were female sex (AOR = 6.4, 95% CI 3.2-12.6); age group of < 35 years (AOR = 23.3, 95% CI 9.0-60.3); lack of regular exercise (AOR = 4.9, 95% CI 2.4-10.2) and working for more than 48 hours per week (AOR = 12.7, 95% CI 6.2 26.2). CONCLUSION: This study reveals that prevalence of anxiety and depression among family practitioners in Karachi is high and higher than general population in Karachi. Further research and intervention studies are required to identify preventive measures in this regard and also to assess the impact of these interventions. PMID- 15134202 TI - Spontaneous recovery from depression in women: a qualitative study of vulnerabilities, strengths and resources. AB - OBJECTIVE: To gain insight into the perceived vulnerability and restitution factors for anxiety/or depression. METHODS: Focus group discussion of seven married women recovered spontaneously from anxiety and/or depression, belonging to a lower middle class semi-urban community of Karachi. RESULTS: Poverty, unemployment, abuse and on going difficulties were perceived as risk factors for depression. A reliable social support system, positive thinking approach, faith, prayers, and experiencing a "turning point" event were reported as factors that promoted recovery from anxiety and/or depression. CONCLUSION: Individual vulnerabilities, strengths and resources can have an important role in recovery from anxiety and/or depression in women. PMID- 15134203 TI - Q-probes study of replicate specimens at the clinical laboratory, the Aga Khan University Hospital, Karachi. AB - OBJECTIVE: The aim of this Q-probes study was to evaluate the precision of replicate specimens. METHODS: This Q-probes study done at the Clinical Laboratory, The Aga Khan University Hospital Karachi is about routine chemistry analytes because of their importance in critical care patients. The analytes data was collected for six months from April to September 2002. There were total 358 samples, which were given for reanalysis during this period under the constant working conditions. After analysis, results of both the runs on different days, were compared to determine the percent difference between the results. After calculating the percent difference, the results were either accepted or rejected on the basis of guidelines set by CLIA 88(Clinical Laboratories Improvement Amendment 1988). RESULTS: Among 358 results, 5 were rejected according to the criteria selected. All of these results were then subjected to statistical analysis for calculating statistical significance. As our Null hypothesis was that the prevalence of rejected results were more than 2% with an acceptable limit of less than or equal to 2%. We failed to accept the Null hypothesis that means that it did not exceed the acceptable limits (p-value = 0.962930) with the Confidence Interval range of 0.2 -2.6 where Upper Confidence Interval is still less than 5% for a p-value of 0.05. The rejection of null hypothesis favors high precision between the two sets of results in our studied population. CONCLUSION: It is concluded that Q-Probes study of replicate specimens at the Clinical Laboratory at The Aga Khan University Hospital is within the acceptable limits. These figures show that a high quality precision is maintained among the observed specimens. This is an ongoing exercise and studies like these should be a continuous process to maintain and enhance the quality of Laboratory results). PMID- 15134204 TI - Prevalence of chronic complications and associated factors in type 2 diabetes. AB - OBJECTIVE: To determine the prevalence of chronic complications and associated factors in type 2 diabetes in 500 diabetic patients, age > or = 25 years, attending the clinic of Diabetic Association of Pakistan (DAP), Karachi. METHODS: Every 5th registered diabetic patient > or = 25 years age, was examined for the presence/absence of micro and macro vascular complications and associated factors. Blood samples were collected for HbA1c, lipid profile and serum creatinine. Urine was examined for albumin and microalbumin. RESULTS: Of the 500 diabetic patients examined (160 males, 340 females, mean age 55.2 10.6 years), retinopathy was seen in 43%, neuropathy in 39.6% and foot ulcers in 4%. Nephropathy was found in 20.2%, and was significantly associated with hypertension. The prevalence of microvascular complications was higher in the group of patients with HbA1c > 8% and was significantly related to duration of diabetes, hypertension and obesity. Hypertension was manifest in 64.6% patients, 61% had raised Body Mass Index and Waist Hip Ratio was more than normal in 88% subjects. Macrovascular complications were encountered in 102 diabetic patients, with angina in 85 (17%), heart attack in 25 (5%) and stroke in 13 (2.6%). CONCLUSION: The prevalence of diabetic microvascular complications was higher in people with poor glycaemic control, longer duration of diabetes and associated hypertension and obesity. PMID- 15134205 TI - Macrovascular complications and their associated factors among persons with type 2 diabetes in Karachi, Pakistan--a multi-center study. AB - OBJECTIVE: To assess the prevalence of macrovascular complications and factors associated with these complications among persons with Type 2 diabetes. METHODS: A multi-center, cross-sectional survey was conducted in three diabetes clinics of Karachi from November 2000 to April 2001. Six hundred and seventy-two persons were interviewed to determine the prevalence of ischemic heart disease (IHD), cerebrovascular disease (CVA) and diabetic foot (DF). Demographic data and co variables obtained include age, sex, duration of diabetes, body mass index (BMI), smoking status, exercise habits and history of hypertension (HTN). RESULTS: Overall, 26.4%, 6.8% and 3.9% of individuals had IHD, CVA and DF respectively. Multiple logistic regression analysis showed that subjects having IHD were more likely to have HTN (AOR=1.88, 95% CI 1.31-2.69), not performing regular exercise (AOR = 2.09, 95% CI 1.39-3.17) and be current smokers (AOR = 1.85, 95% CI 1.10 3.10) Subjects having CVA were more likely to have diabetes for more than 5 years (AOR = 1.94, 95% CI 0.97-3.87). Males were more prone than females to have DF (AOR = 3.48, 95% CI 1.46-8.31). Those who had DF were more likely to have diabetes for more than 5 years (AOR = 3.29, 95% CI 1.09-9.89) and of reporting current smoking (AOR = 4.01, 95% CI 1.49-10.71). CONCLUSION: Large proportions of persons with Type 2 diabetes were suffering from preventable macrovascular complications in Karachi, Pakistan. There is a need to develop risk factor modification interventions to reduce the impact of long-term complications. PMID- 15134206 TI - Baseline disease knowledge assessment in patients with type 2 diabetes in a rural area of northwest of Pakistan. AB - OBJECTIVE: (1) To assess baseline disease related knowledge in patients with type 2 diabetes about their disease, its risk factors, signs/symptoms, related complications and suitable diet and (2) Is there an association between gender, duration of disease & age at diagnosis of diabetes and the above dependent variables. METHODS: A 20-item interview-based structured knowledge questionnaire was used to collect information. A total of 82, diabetic patients, mean age 55.2 (11.4 S.D.) years, ranging from 35-80 years, were interviewed. RESULTS: Statistically significant association was found between age at diagnosis and better understanding of risk factors, (OR = 1.20, P = 0.012 with 95% CI 0.85 - 0.98). Statistically significant association was found between gender and better understanding of word "diabetes" or "sugar" OR = 1.15, P = 0.051 with 95% Confidence interval 0.96-1.29). Statistically significant associations were found between gender and patients' better understanding of disease signs/symptoms (OR = 1.35, P = 0.005 with 95% CI 0.40-0.56). No significant associations were found between gender, duration of diabetes, age at diagnosis and patients' better understanding of disease related complications and suitable diet in diabetes. Duration of diabetes was not statistically associated with any of the dependent variables. CONCLUSION: Priority needs to be given by WHO education programmes for the development of diabetes education program in rural areas to give patients a better knowledge of their disease, to prevent premature morbidity and mortality associated with diabetes. PMID- 15134207 TI - Survival after myocardial infarction in patients with type 2 diabetes. AB - OBJECTIVES: To determine prospectively the survival of previously known diabetic patients admitted to coronary care unit with confirmed myocardial infarction (MI) over a one-year period and to assess the effects of gender, age, diabetes duration, hyperglycemia, hypertension, and anti-diabetic treatment on survival. METHODS: In this prospective cohort study, we followed 59 patients hospitalized with a confirmed myocardial infarction at 3 coronary care units, Peshawar (Pakistan), between May 1, 2000 and April 30, 2001. We analyzed survival using univariate and multivariate Cox Proportional hazards regression models to control for potentially confounding factors. RESULTS: A total of 17 (28.8%) subjects (7 male and 10 female) died. Survival was significantly associated with previous history of hypertension and duration of diabetes (HR = 3.40, 95% CI = 1.33-9.22, P = 0.001, and HR = 1.24, 95% CI = 1.05-1.45, P = 0.009, by Univariate Cox model, respectively). In multivariate analysis duration of diabetes was significantly associated with survival and Metformin treatment only, and Sulphonylurea & Metformin treatment together (P = 0.003, and 0.026, Multivariate Cox model, respectively). CONCLUSION: Our results suggested that diabetes is associated with markedly increased mortality in the presence of hypertension after acute myocardial infarction and support for aggressive treatment of coronary risk factors among diabetic patients. PMID- 15134208 TI - Placenta previa: the role of ultrasound in assessment during third trimester. AB - OBJECTIVE: To assess the role of ultrasound in detecting the migration of placenta previa during the third trimester at Lady Willingdon Hospital and Jinnah Hospital, Lahore during the period July 2000 to September 2002. METHODS: Eighty pregnant women with the diagnosis of placenta previa at 28 to 32 weeks of gestation were included in the study. After base line ultrasound, scan was repeated every two weeks until delivery or placental migration for more than 3 cm from internal cervical os. Detailed information for placental position, distance from cervical os and relation to presenting part was recorded. Women with major degree placenta previa were admitted in the hospital at 32 -34 weeks of gestation. Delivery plan was made according to degree of placenta previa by completed 37 weeks of gestation. Cesarean section was done for the women with major degree placenta previa and minor degree placenta previa with antepartum hemorrhage and obstetric indication RESULTS: Out of 80 women placental migration to a distance of more than 3-5 cm from the internal cervical os occurred in 20 cases (12 anterior/anterolateral, 8 posterior/posterolateral) by 36 weeks of gestation and 20 had complete placenta previa. Out of remaining 40 cases, 12 patients had vaginal delivery and 28 had cesarean section. Placental migration was not observed in women with total placenta previa or posterior placenta previa when the distance of lower edge of placenta was less than 1 cm from the internal os. CONCLUSION: Ultrasound is important for the diagnosis of placental localization and placental migration during third trimester. Placental migration takes place more often in anterior than in complete or posterior placenta previa. PMID- 15134209 TI - Prognostic indicators in patients with intracranial tuberculoma: a review of 102 cases. AB - OBJECTIVE: To see the characteristics, course and outcome of patients suffering from intracranial tuberculoma. METHODS: Retrospective review of 102 patients diagnosed as intracranial tuberculoma at a tertiary care center over 10 years. RESULTS: A total of 102 cases were seen with an age range of 1 to 75 years (mean, 30 years). Predisposing factors included Diabetes mellitus (8 patients) and pregnancy or puerperium (7 patients). Five pediatric patients had tuberculoma despite documented BCG vaccination. Fever (59%), headache (57%), meningeal irritation (36%) were the commonest presenting features; one-third of patients were drowsy or comatosed at presentation. Cerebrospinal fluid analysis was performed in 63 patients, of whom 88% had elevated protein, 83% had low glucose, and 84% had pleocytosis (one-third with neutrophilia). Forty-nine (50%) patients had clinical or laboratory evidence of concomitant tuberculous meningitis. Chest radiographs showed active or old tuberculous infection (25%), with a miliary pattern in 20%. Two-thirds of subjects had multiple tuberculomas (mean, 4.5 lesions per patient) on contrast CT or MRI scan. Hydrocephalus was present in 37 (37%) patients of which 21 required shunt surgery. Thirty-nine patients had > 9 months of follow up; 17 patients showed complete recovery, 20 patients had partial recovery, and 2 patients had no response. Coma at presentation and miliary pattern on chest X-ray were predictors of poor prognosis. CONCLUSION: The study demonstrate that fever, headache, signs of meningeal irritation and cranial nerve palsies are common presenting features. Complete recovery was seen in 40% patients. Coma and military TB are predictors of poor prognosis. PMID- 15134210 TI - Empirical treatment for tuberculosis: survey of cases treated over 2 years in a London area. AB - OBJECTIVE: To determine factors influencing the decision to treat tuberculosis empirically and its outcome. METHODS: A retrospective survey was done to assess the factors influencing the decision to treat tuberculosis empirically and the effectiveness of such treatment, within a London area. Data on tuberculosis cases treated during 1995-96 (n = 218) was collected and analysed. RESULTS: One hundred and fifty-eight (72.5%) cases of tuberculosis were treated empirically, that is, in the absence of positive smear or histology. Factors suggesting tuberculosis were clinical suspicion, abnormal radiology, elevated Erythrocyte Sedimentation Rate (ESR) and/or C-reactive Protein (CRP), grade 3 or 4 Heaf test, contact or family history, and Heaf conversion. Significantly more Asian [79 of 105 (75.2%); p < 0.02] and African patients [61 of 81 (75.3%); p < 0.05] were treated empirically as compared to West European patients [14 of 28 (50%)]. Based on logistic regression, any non-European ethnicity carried a significant risk for being treated empirically [OR: 2.7, CI: 1.7-20.8; p < 0.05]. Patients requiring revision of diagnosis (n = 9) were older [55.7 +/- 10.1 vs 32.8 +/- 16.0 years; p < 0.001]; 6 of them had neoplastic conditions. CONCLUSION: In this survey, frequent use of empirical treatment for tuberculosis in non-European patients reflects the physicians' high index of suspicion. Although such therapy is justifiable in selected cases, the proportion requiring revision of diagnosis may be reduced by more thorough investigation, especially in older patients. PMID- 15134211 TI - Prevalence of exercise-induced bronchospasm in national hockey players of Pakistan. AB - OBJECTIVE: To determine prevalence of Exercise-induced Bronchospasm (EIB) in hockey players, who had represented or were aspiring to represent Pakistan at international level. METHOD: An observational, cross-sectional study was done on the prevalence of EIB in national hockey players of Pakistan. All participants of the training camp (n = 27) players were included in this study, after obtaining permission from the Pakistan Hockey Federation. An acquaintance session was conducted to introduce the study, and for the correct recording of Peak Expiratory Flow (PEF) rate. On field-testing day, pulse and PEF rates were measured first without any exercise and warm up (pre-exercise PEF rate). After competitive 6 minute (6-min) field free running, pulse rate was measured to ensure that players heart rate has reached 70% of the predicted maximum heart rate. PEF and pulse rates were measured at 5-min, 15-min and 30-min post exercise. A player was considered EIB positive based on a post exercise decrement in PEF rate > or = 15% at any defined point of time. RESULTS: Twenty-seven players participated in our study out of which 5 players (19%) had EIB. CONCLUSION: Our findings indicate that a significant number of cases of EIB exist among national field hockey players. We suggest that active screening for EIB should be made part of training sessions, so that performance of players could be enhanced. Diagnosed cases should be treated as per the guidelines laid down by International Olympic Committee Medical Commission. PMID- 15134212 TI - Banti's syndrome: case report and review of literature. PMID- 15134213 TI - Localized small bowel perforation--a radiological rarity. PMID- 15134214 TI - Fatal case of chickenpox in an adult: a case report. PMID- 15134215 TI - Administering painless injections. PMID- 15134216 TI - Intractable cancers: the many faces of multidrug resistance and the many targets it presents for therapeutic attack. AB - Some types of cancer respond far less favorably to treatment than do others. A quantitative estimate of this intuition can be obtained from the SEER (Surveillance, Epidemiology and End-Results) Cancer Statistics Review. Of particular interest, from a drug resistance perspective, are the five-year survival data for patients presenting with tumors that were diagnosed as "distant". A good correlation can be found between those numbers and an estimate of treatment successes obtained from a survey of current literature on chemotherapy applied to cancers originating from these various tissues. These two measures, considered together, define "the axis of intractability", a parameter that characterizes the (possibly) inherent, physiological basis of the tissue-by tissue intractability of cancers. Exploring the basis of this intractability, it appears that factors other than the classical ABC transporter-based, multidrug resistance systems probably play a major role. An ineffective DNA repair system, coupled to reduced apoptosis, is the basis for the inherent tractability of testicular cancer. For other tissues, important contributions to resistance arise from cell adhesion-mediated drug resistance, which is overcome when cells are released from tissues during anoikis. Making a direct comparison between gene expression in solid tumors and their corresponding cell lines, genes controlling the extracellular matrix and cell-cell communication appear among the genes that are over-expressed in the solid tumors, while genes coding for the protein biosynthesis system are over-expressed in the cell lines. The more tractable cancers are closer to the cell lines in their expression profiles of these sets of genes. PMID- 15134217 TI - Cellular targets for anticancer strategies. AB - Since late 1950s the main strategies to treat cancer, besides surgery, have been radiotherapy or chemotherapy. These approaches work primarily by damaging proliferating cells at the level of DNA replication or cell division, and inducing apoptotic cell suicide as a secondary response to the damage. In recent years, efforts to improve cancer therapy have focused on the development of more selective, biological mechanism based approaches that can help to overcome tumor resistance as well as minimize toxic side effects. In the present review new strategies and new targets for biological cancer therapy will be discussed. In particular, new angiogenic pathways discovered in melanoma will be discussed in relationship to a more efficient anticancer strategy. In summary, this review tries to identify the most logical targets and the most useful mechanisms of tumor inhibition in light of new knowledge from the basic research including human genome project. PMID- 15134218 TI - DNA-binding properties of poly(ADP-ribose) polymerase: a target for anticancer therapy. AB - Poly(ADP-ribose) polymerization is a unique post-translation protein modification that utilizes an ADP-ribose moiety from NAD+ to form long and branched polymers attached via glutamic acid residues to nuclear acceptor proteins. The corresponding enzyme, poly(ADP-ribose) polymerase (PARP-1), is a zinc finger containing protein, which allows PARP-1 binding to either double- or single strand DNA breaks. The catalytic activity of PARP-1 is strictly dependent on the presence of strand breaks in DNA, and is modulated by the level of automodification. PARP-1 is regarded as an intracellular sensor for DNA strand breaks, and its function has been implicated in cellular processes that require DNA cleavage and rejoining reactions, such as DNA replication, recombination and repair. Recent studies have also implicated PARP-1 in the regulation of gene expression through modification of transcription factors by poly(ADP ribosyl)ation or its direct binding to gene-regulating DNA sequences. The latter is attributable to PARP's ability to recognize and bind to various structural discontinuities in the DNA duplex in the absence of DNA strand breaks, such as three- or four-way junctions, bent DNA, and base unpaired regions. Cumulatively, these findings indicate that PARP-1 plays a pivotal role in the maintenance of the genome integrity during the normal functioning of eukaryotic cells as well as in the cellular responses to DNA damage, and that PARP-DNA interactions are indispensable for PARP function. This review summarizes the data on DNA-binding properties of PARP-1 and relates them to the development of strategies for sensitizing tumor cells to genotoxic treatments. PMID- 15134219 TI - CARD proteins as therapeutic targets in cancer. AB - Proteins containing a caspase-associated recruitment domain (CARD) have been established as key regulators of cell death and, more recently, cytokine production. During the last several years, the number of proteins identified within this family has grown immensely and many aspects of their function point to their potential utility as novel drug targets in the treatment of cancer. Several CARD family proteins are critical components of the conserved cell death machinery which, when dysregulated, promotes oncogenesis and contributes prominently to tumor resistance to chemotherapy. The pro-apoptotic protein Apaf1, which is inactivated in some cancers, is a CARD protein that is indispensable for mitochondria-induced apoptosis. Other anti-apoptotic CARD proteins, such as TUCAN/CARDINAL/CARD8, have been shown to protect tumors from cell death stimuli and to be over-expressed in certain forms of cancer. Therapeutics that activate or inhibit CARD proteins may therefore be potentially utilized as novel chemo sensitizing agents when used in conjunction with conventional chemotherapy. Other CARD proteins influence cellular processes through the regulation of NF-kappaB or caspase-1, which governs the levels of interleukin-1beta (IL-1beta). In addition to its pro-inflammatory properties, this cytokine also contributes to neoplastic progression by promoting angiogenesis, proliferation, and the metastasis of many tumors. Many of the IL-1beta-regulating CARD proteins also contain a nucleotide binding/oligomerization domain known as a NACHT and may therefore be amenable to targeting by small molecule compounds. This review examines the role of CARD proteins in cytoprotection and cytokine processing in the context of neoplasia and presents strategies for using this information in devising potential novel anticancer agents. PMID- 15134220 TI - Sphingolipid metabolism enzymes as targets for anticancer therapy. AB - Treatment with anti-cancer agents in most cases ultimately results in apoptotic cell death of the target tumor cells. Unfortunately, tumor cells can develop multidrug resistance, e.g., by a reduced propensity to engage in apoptosis by which they become insensitive to multiple chemotherapeutics. Ceramide. the central molecule in cellular sphingolipid metabolism, has been recognized as an important mediator of apoptosis. Moreover, an increased cellular capacity for ceramide glycosylation has been identified as a novel multidrug resistance mechanism. Indeed, virtually all multidrug resistant cell types exhibit a deviating sphingolipid composition, most typically an increased level of glucosylceramide. Thus, the enzyme glucosylceramide synthase, which converts ceramide into glucosylceramide, has emerged as a potential target to increase apoptosis and decrease drug resistance of tumor cells. In addition, several other steps in the pathways of sphingolipid metabolism arc altered in multidrug resistant cells, opening a perspective on additional sphingolipid metabolism enzymes as targets for anti-cancer therapy. In this article, we present an overview of the current understanding concerning drug resistance-related changes in sphingolipid metabolism and how interference with this metabolism can be exploited to over come multidrug resistance. PMID- 15134221 TI - Potential for predicting toxicity and response of fluoropyrimidines in patients. AB - The efficacy of cancer therapy is compromised by the fact that there are currently no good ways to predict which patients will benefit from treatment. This long standing goal is closer to becoming a reality as more is learned about the molecules that affect the activities of various therapeutic agents. The fluoropyrimidine antimetabolites drugs have been in clinical use for over 4 decades and the cellular proteins important for their activities have been studied in detail. The most important are the major target enzyme, thymidylate synthase (TS) and the rate limiting enzyme in the degradation pathway, dihydropyrimidine dehydrogenase (DPD), equally important for the analogue capecitabine is thymidine phosphorylase (TP), which is rate limiting for activation of this prodrug. A number of assays are available for these enzymes, including enzyme activity measurements. quantitative PCR for RNA expression and immunological methods for protein expression. With each of these methods, more clinical studies are required to validate their clinical usefulness. PMID- 15134222 TI - Molecular targeting of drug delivery systems to cancer. AB - This review presents molecular targeting approaches in anticancer drug delivery systems (DDS) and identifies new developments in these systems. Targeting approaches include passive targeting (enhanced permeability and retention effect), targeting specific tumor conditions, topical delivery and active targeting, namely, targeting organs, cells, intracellular organelles and molecules, sandwich targeting, promoter targeting, indirect targeting and targeting by external stimuli. A novel advanced proapoptotic anticancer DDS that utilizes several molecular targets will be considered. Experimental data suggest that this DDS can simultaneously: (1) induce cell death, (2) prevent adverse effects on healthy tissues; (3) suppress and prevent multidrug resistance; and (4) inhibit cellular antiapoptotic defense. PMID- 15134223 TI - [Urinary tract infections in children]. AB - The diagnosis of urinary tract infections is based on the number of colony forming units that grow on the culture media. The criteria for the diagnosis depends on the method of collection: clean void and bags > 10(5)/mL; transurethral catheterization > 10(4)/mL. The diagnosis of acute pyelonephritis or cystitis is based on clinical and biological signs: unexplained fever, abdominal pains, signs of neonatal infections, elevated white blood cell count, C reactive or sedimentation rate. After an urinary tract infection, it is important to look for a lithiasis, an obstructive uropathy, a vesico-ureteral reflux and a bladder dysfunction. The child should have a sonogram and a voiding cystourethrography. In case of cystitis the antimicrobial therapy is given orally. In case of pyelonephritis, most of the time the treatment is initially administered parenterally for 3 days and then completed by a 10-day antimicrobial course orally. In a child over 18 months of age who is not toxic appearing without severe uropathy, the antimicrobial therapy can be initiated orally. PMID- 15134224 TI - [Implantable cardioverter defibrillators]. PMID- 15134225 TI - [Arrhythmias and emergencies. Considering step by step]. PMID- 15134226 TI - [Narrow QRS tachycardia]. AB - Narrow QRS tachycardia are supraventricular tachycardia, atrial tachycardia or junctional tachycardia. The diagnosis is done with the ECG, after identification of atrial activation, eventually after vagal manoeuvres, and transoesophageal recordings. DC Choc, atrial pacing, or antiarrhythmic drugs can do interruption. All atrial tachycardia must be anticoagulated prior regularisation. There is seldom an emergency for regularisation, as drugs can always slow the ventricular rate down. Prevention of relapses uses oral antiarrhythmic drugs, sometimes pacing. Most of these tachycardia can be cured by catheter ablation, essentially flutter and junctional tachycardia. PMID- 15134227 TI - [Atrial fibrillation]. AB - Atrial fibrillation is a common arrhythmia which consequences include disabling symptoms, haemodynamic impairment and frightening embolic complications. In 3/4 cases, they are represented by cerebrovascular accidents responsible of death or disabling sequella. Underlying heart disease is present in 70% of AF patients. Endpoints of therapy include: 1. prevention of embolic complications using oral anticoagulation in patients at risk; 2. control of symptoms and prevention of haemodynamic impairment either by restoring and maintaining sinus rhythm or by controlling ventricular heart rate. The same principles should be applied for emergency treatment: aside from AF with haemodynamic compromise (hypotension or syncope) which requires urgent electrical cardioversion, AF termination may be obtained with intravenous or oral antiarrhythmic therapy or the treatment confined to slowing of heart rate. Selecting the appropriate antiarrhythmic therapy for prevention of recurrences is based on the type of AF, paroxysmal or persistent, symptoms, underlying heart disease and left ventricular function. For patients refractory to drug therapy, non pharmacologic treatment represents an option including pacemaker, double chamber defibrillator, and radiofrequency ablation of ectopic foci most often located in the pulmonary veins or pulmonary vein isolation or surgery particularly if there is an indication for open-chest surgery. Non-pharmacological therapies should be restricted to very symptomatic patients who failed pharmacological therapy. PMID- 15134228 TI - [Atrial arrhythmia: when should a heart stimulator be implanted?]. PMID- 15134229 TI - [Typical and atypical atrial flutters]. AB - Definitions, classification, surface electrocardiographic characterisitics, risk for stroke and treatments. Atrial flutters are macro-reentrant circuits originated from both right and left atria. They have been classified in typical and atypical based on their cavo-tricuspid isthmus-dependence. Typical atrial flutter comprises counterclockwise and clockwise isthmus-dependent atrial flutter located within the right atrium. Atypical atrial flutters are composed by non isthmus-dependent right atrial flutter and left atrial flutter. Typical atrial flutter represents at least 90% of all flutters, and has usually a counterclockwise pattern. Ablation is a potential definitive therapy for atrial flutter. However, successful procedures as well as complications rates are different depending of the location of the circuit. There are now evidences for risk of stroke related to atrial flutter, although epidemiological studies have not been conducted yet. Therefore, prevention of such complications using anti thrombotic should be performed as done for atrial fibrillation. PMID- 15134230 TI - [Management of wide QRS complex tachycardia]. AB - Wide QRS complex tachycardias present significant diagnostic and therapeutic challenges to emergency physicians, cardiologists, anesthaesiologists, and intensive care doctors. Wide QRS complex tachycardias have to be initially considered as ventricular tachycardia before any other investigation even if this origin is supraventricular. The first step is to determine the tolerance of the tachycardia. If the tachycardia is associated with syncope, cardiac arrest, severe hypotension or angina, DC cardioversion is mandatory. If the tachycardia is well tolerated, the bedside diagnosis should take into account the clinical context, clinical history and analysis of the surface 12 leads ECG. This article presents the diagnostic and therapeutic approaches to wide ORS complex tachycardias. PMID- 15134231 TI - [Diagnostic criteria for ventricular tachycardia]. PMID- 15134232 TI - [Wolff-Parkinson-White syndrome]. PMID- 15134233 TI - [Arrhythmic cardiomyopathy of the right ventricle: risk of sudden death in young subjects]. PMID- 15134234 TI - [A risk of sudden death to be screened: Brugada's syndrome]. PMID- 15134235 TI - [Prevention of cardiac arrhythmias: what is the place for antiarrhythmic drugs?]. AB - The place of antiarrhythmic drugs in the preventive treatment of the cardiac arrhythmias was recently revalued because of the improvement of the radiofrequency catheter ablation techniques and the growing place of the implantable cardioverter defibrillator. The choice of the antiarrhythmic treatment must always be based on the type of arrhythmia, the presence of a structural cardiopathy and the potential side effects of the antiarrhythmic drug. PMID- 15134236 TI - [Implantable automatic defibrillator: challenges and delays in France]. PMID- 15134237 TI - [Ablative strategy: a definite treatment for cardiac arrhythmias?]. AB - The development of radiofrequency catheter ablation has dramatically changed the therapeutic approach of cardiac arrhythmias. All atrial rhythm disturbances are now amenable to ablative strategy. Ablation has become the first-line therapy for atrial flutter. Atrial fibrillation, a major factor of morbidity and mortality, is now a target for this radical treatment. Ablation of pulmonary vein foci, eventually associated to atrial linear lesions, raises the possibility of suppressing atrial fibrillation. The success rate is now as high as 70 to 85%, depending on the form of atrial fibrillation. For atrioventricular reentrant tachycardia either nodal or over an accessory pathway, ablation is the reference treatment with success rates of 99%. All symptomatic Wolff-Parkinson-White syndromes can be definitely cured by this technique. At the ventricular level, the procedures are often more complexes, due to advanced cardiomyopathy. However, most of sustained ventricular tachycardias can be cured by ablation. The success rates are somewhat lower (about 80%). The challenge for the next years is ablation of premature ventricular beats initiating ventricular fibrillation. The technique is also improving from day-to-day (88% success rate) and makes rise the hope of curing sudden death. PMID- 15134238 TI - [Early defibrillation and sudden death: the French delay]. PMID- 15134239 TI - [Antithrombotic therapy in atrial arrhythmia]. AB - The principal complication of the atrial arrythmias is the thrombo-embolic accident, notably the cerebro-vascular accident. The efficacity of the oral anticoagulants in reducing cerebro-vascular accidents has been demonstrated in numerous studies. This is significantly superior to that obtained with the anti platelet drugs. However, the anti-vitamin K drugs (warfarin) carry a risk of serious haemorrhage of around 5% per year. This restricts the proposal of this treatment to patients with an elevated risk of vascular accidents: age, diabetes, previous cerebro-vascular accidents, and cardiac failure are the risk factors. Nevertheless, the risk of haemorrhage is responsible for an under prescription of the anticoagulants in the elderly. This explains the interest aroused by alternative therapeutics: the results of trials on ximelagatran, a direct anti thrombin, are promising. In patients with an arrythmia, cardioversion carries a thrombo-embolic risk of around 1%. This risk is reduced by prior anticoagulant treatment. The procedure for this treatment is orientated by a trans-oesophageal echocardiogram. The incertitude of the duration of anticoagulant therapy without cardioversion calls for respect of the arrythmia. The treatment of this is limited to control of the cardiac rhythm and anticoagulant treatment. PMID- 15134240 TI - [Patient card. Tips on taking an oral anticoagulant (antivitamin K)]. PMID- 15134241 TI - [Assessment, correcting, weighting, rating, ranking...]. PMID- 15134242 TI - [Mental handicap. Administrative supervision, trusteeship, safeguard of justice]. PMID- 15134243 TI - [Evaluation of the severity, and search of early complications in limb injuries]. PMID- 15134244 TI - [The placebo effect and placebo medications]. PMID- 15134245 TI - [Pharmaceutical and non-pharmaceutical therapeutics. Regulation and recommendations]. PMID- 15134246 TI - [Larrey, a legendary surgeon, a current work]. PMID- 15134247 TI - Viral control of phytoplankton populations--a review. AB - Phytoplankton population dynamics are the result of imbalances between reproduction and losses. Losses include grazing, sinking, and natural mortality. As the importance of microbes in aquatic ecology has been recognized, so has the potential significance of viruses as mortality agents for phytoplankton. The field of algal virus ecology is steadily changing and advancing as new viruses are isolated and new methods are developed for quantifying the impact of viruses on phytoplankton dynamics and diversity. With this development, evidence is accumulating that viruses can control phytoplankton dynamics through reduction of host populations, or by preventing algal host populations from reaching high levels. The identification of highly specific host ranges of viruses is changing our understanding of population dynamics. Viral-mediated mortality may not only affect algal species succession, but may also affect intraspecies succession. Through cellular lysis, viruses indirectly affect the fluxes of energy, nutrients, and organic matter, especially during algal bloom events when biomass is high. Although the importance of viruses is presently recognized, it is apparent that many aspects of viral-mediated mortality of phytoplankton are still poorly understood. It is imperative that future research addresses the mechanisms that regulate virus infectivity, host resistance, genotype richness, abundance, and the fate of viruses over time and space. PMID- 15134248 TI - Algicidal bacteria in the sea and their impact on algal blooms. AB - Over the past two decades, many reports have revealed the existence of bacteria capable of killing phytoplankton. These algicidal bacteria sometimes increase in abundance concurrently with the decline of algal blooms, suggesting that they may affect algal bloom dynamics. Here, we synthesize the existing knowledge on algicidal bacteria interactions with marine eukaryotic microalgae. We discuss the effectiveness of the current methods to characterize the algicidal phenotype in an ecosystem context. We briefly consider the literature on the phylogenetic identification of algicidal bacteria, their interaction with their algal prey, the characterization of algicidal molecules, and the enumeration of algicidal bacteria during algal blooms. We conclude that, due to limitations of current methods, the evidence for algicidal bacteria causing algal bloom decline is circumstantial. New methods and an ecosystem approach are needed to test hypotheses on the impact of algicidal bacteria in algal bloom dynamics. This will require enlarging the scope of inquiry from its current focus on the potential utility of algicidal bacteria in the control of harmful algal blooms. We suggest conceptualizing bacterial algicidy within the general problem of bacterial regulation of algal community structure in the ocean. PMID- 15134249 TI - Parasites and phytoplankton, with special emphasis on dinoflagellate infections. AB - Planktonic members of most algal groups are known to harbor intracellular symbionts, including viruses, bacteria, fungi, and protozoa. Among the dinoflagellates, viral and bacterial associations were recognized a quarter century ago, yet their impact on host populations remains largely unresolved. By contrast, fungal and protozoan infections of dinoflagellates are well documented and generally viewed as playing major roles in host population dynamics. Our understanding of fungal parasites is largely based on studies for freshwater diatoms and dinoflagellates, although fungal infections are known for some marine phytoplankton. In freshwater systems, fungal chytrids have been linked to mass mortalities of host organisms, suppression or retardation of phytoplankton blooms, and selective effects on species composition leading to successional changes in plankton communities. Parasitic dinoflagellates of the genus Amoebophrya and the newly described Perkinsozoa, Parvilucifera infectans, are widely distributed in coastal waters of the world where they commonly infect photosynthetic and heterotrophic dinoflagellates. Recent work indicates that these parasites can have significant impacts on host physiology, behavior, and bloom dynamics. Thus, parasitism needs to be carefully considered in developing concepts about plankton dynamics and the flow of material in marine food webs. PMID- 15134250 TI - Interactions between planktonic microalgae and protozoan grazers. AB - For an algal bloom to develop, the growth rate of the bloom-forming species must exceed the sum of all loss processes. Among these loss processes, grazing is generally believed to be one of the more important factors. Based on numerous field studies, it is now recognized that microzooplankton are dominant consumers of phytoplankton in both open ocean and coastal waters. Heterotrophic protists, a major component of microzooplankton communities, constitute a vast complex of diverse feeding strategies and behavior which allow them access to even the larger phytoplankton species. A number of laboratory studies have shown the capability of different protistan species to feed and grow on bloom-forming algal species. Because of short generation times, their ability for fast reaction to short-term variation in food conditions enables phagotrophic protists to fulfill the function of a heterotrophic buffer, which might balance the flow of matter in case of phytoplankton blooms. The importance of grazing as a control of microalgae becomes most apparent by its failure; if community grazing controls initial stages of bloom development, there simply is no bloom. However, if a certain algal species is difficult to graze, e.g. due to specific defense mechanisms, reduced grazing pressure will certainly favor bloom development. The present contribution will provide a general overview on the interactions between planktonic microalgae and protozoan grazers with special emphasis on species specific interactions and algal defense strategies against protozoan grazers. PMID- 15134251 TI - Controlling harmful algal blooms through clay flocculation. AB - The potential use of clays to control harmful algal blooms (HABs) has been explored in East Asia, Australia, the United States, and Sweden. In Japan and South Korea, minerals such as montmorillonite, kaolinite, and yellow loess, have already been used in the field effectively, to protect fish mariculture from Cochlodinium spp. and other blooms. Cell removal occurs through the flocculation of algal and mineral particles, leading to the formation of larger aggregates (i.e. marine snow), which rapidly settle and further entrain cells during their descent. In the U.S., several clays and clay-rich sediments have shown high removal abilities (e.g. > 80% cell removal efficiency) against Karenia brevis, Heterosigma akashiwo, Pfiesteria piscicida and Aureococcus anophagefferens. In some cases, the removal ability of certain clays was further enhanced with chemical flocculants, such as polyaluminum chloride (PAC), to increase their adhesiveness. However, cell removal was also affected by bloom concentration, salinity, and mixing. Cell mortality was observed after clay addition, and increased with increasing clay concentration, and prolonged exposure to clays in the settled layer. Mesocosm, field enclosure, and flume experiments were also conducted to address cell removal with increasing scale and flow, water-column impacts, and the possible benthic effects from clay addition. Results from these studies will be presented, especially those in regards to water quality, seawater chemistry, bottom erodibility and faunal impacts in the benthos. At this time, clay dispersal continues to be a promising method for controlling HABs and mitigating their impacts based on existing information and experimental data. PMID- 15134252 TI - Unexpected foraminiferal diversity revealed by small-subunit rDNA analysis of Antarctic sediment. AB - Studies of benthic Foraminifera typically rely on the morphological identification of dried specimens. This approach can introduce sampling bias against small, delicate, or morphologically ambiguous forms. To overcome this limitation, we extracted total DNA from sediment followed by PCR using group- and species-specific primers. Phylogenetic analyses revealed that approximately ninety percent of the PCR products represented previously undescribed sequence types that group with undersampled members of the allogromiid Foraminifera. We also used a modification of this technique to track individual species in sediment fractions too fine for normal morphological identification, and to confirm species placement of morphologically ambiguous foraminiferans. We were able to identify the DNA of several large foraminiferal species in fine fractions in a seasonally-dependent manner, indicating that in some seasons the majority of the standing stock of these species exists as gametes/juveniles. The approach outlined here represents a powerful strategy for exploring the total diversity of benthic foraminiferal communities. PMID- 15134253 TI - Phylogenetic relationships of the subclass Peritrichia (Oligohymenophorea, Ciliophora) inferred from small subunit rRNA gene sequences. AB - The phylogenetic relationships among peritrichs remain unresolved. In this study, the complete small subunit rRNA (SSrRNA) gene sequences of seven species (Epistylis galea, Campanella umbellaria, Carchesium polypinum, Zoothamnium arbuscula, Vaginicola crystallina, Ophrydium versatile, and Opercularia microdiscum) were determined. Trees were constructed using distance-matrix, maximum-likelihood and maximum-parsimony methods, all of which strongly supported the monophyly of the subclass Peritrichia. Within the peritrichs, 1) E. galea grouped with Opercularia microdiscum and Campanella umbellaria but not the other Epistylis species, which indicates that the genus Epistylis might not be monophyletic; 2) the topological position of Carchesium and Campanella suggested that Carchesium should be placed in the family Zoothamniidae, or be elevated to a higher taxonomic rank, and that Campanella should be independent of the family Epistylididae, and probably be given a new rank; and 3) Opisthonecta grouped strongly with Astylozoon, which suggested that Opisthonecta species were not the ancestors of the stalked peritrichs. PMID- 15134254 TI - Comparative effectiveness of light-microscopic techniques and PCR in detecting Thelohania solenopsae (Microsporidia) infections in red imported fire ants (Solenopsis invicta). AB - The main goal of this study was to compare the effectiveness of three staining techniques (calcofluor white M2R, Giemsa and modified trichrome), and the polymerase chain reaction (PCR) in detecting the microsporidium Thelohania solenopsae in red imported fire ants (Solenopsis invicta). The effect of the number of ants in a sample on the sensitivity of the staining techniques and the PCR, and the effect of three DNA extraction protocols on the sensitivity of PCR were also examined. In the first protocol, the ants were macerated and the crude homogenate was used immediately in the PCR. In the second protocol, the homogenate was placed on a special membrane (FTA card) that traps DNA, which is subsequently used in the PCR. In the third protocol, the DNA was purified from the homogenate by traditional phenol-chloroform extraction. Except for PCR using FTA cards, the sensitivity (number of samples positive for T. solenopsae) of all detection techniques increased with the number of ants in the sample. Overall, Giemsa was the least sensitive of all detection techniques. Calcofluor was more sensitive than modified trichrome with ants from one site and was equally as sensitive as PCR with crude DNA or a FTA card with ants from both sites. Trichrome staining was equally as sensitive as PCR with a FTA card at both sites, but it was less sensitive than PCR with crude DNA at one site. PCR on FTA cards was less sensitive than PCR with crude DNA for ants from one site but not the other. There was no difference whether crude or phenol-chloroform purified DNA was used as template. In summary, the results of this study show that PCR based on a crude DNA solution is equal to or more sensitive in detecting T. solenopsae than the other detection techniques investigated, and that it can be used as a reliable diagnostic tool for screening field samples of S. invicta for T. solenopsae. Nevertheless, ant smear stained with calcofluor or modified trichrome should be used to buttress findings from PCR. PMID- 15134255 TI - Molecular and physiological evaluation of subtropical environmental isolates of Acanthamoeba spp., causal agent of Acanthamoeba keratitis. AB - Previous molecular examination of Acanthamoeba spp. has resulted in the determination of distinct genotypes in this genus (designated T1-T12, T14). Genotype T4 has been responsible for the majority of cases of Acanthamoeba keratitis. Here we examine the relative abundance of environmental T4 isolates on beaches and ask whether they have temperature and salinity tolerances that could enhance pathogenicity. Twenty-four Acanthamoeba strains were isolated from beach sand (n = 20), soil (n = 3), and tap water (n = 1) in south Florida. Phylogenetic analysis identified 19 of 24 isolates as T4, the Acanthamoeba keratitis associated genotype. The remaining isolates were genotype T5 (4) and T11 (1). Nearly all beach isolates were genotype T4, whereas the tap water and soil isolates were mostly T5. All amoebae grew at 0, 1.0, and 2.0% salt and 19 of 20 beach isolates also grew at 3.2%. No soil or tap-water acanthamoebae reproduced at 3.2%. All isolates grew at 37 degrees C and two (T5) at 42 degrees C. Little correlation existed between beach location, salt-tolerance, and genetic relatedness. Overall, the large majority of environmental isolates obtained were genotype T4, suggesting it may be the most common genotype in this environment and could be a potential source of Acanthamoeba keratitis infections. PMID- 15134256 TI - The epiplasm gene EPC1 influences cell shape and cortical pattern in Tetrahymena thermophila. AB - The cortical protein Epc1p is the most abundant protein in the membrane skeleton, or epiplasm, of Tetrahymena thermophila. A partial sequence of the EPC1 gene was obtained and used to obtain a knockout construct that was successful in transforming Tetrahymena thermophila cells. The results support the conclusion that Epc1p influences cell shape and the fidelity of cortical development. It was further observed that this protein is transferred from plus to minus cells during conjugation, and that the imported protein is assembled into the epiplasm of the recipient cell in a discreet series of steps. PMID- 15134257 TI - Transfer of Nosema locustae (Microsporidia) to Antonospora locustae n. comb. based on molecular and ultrastructural data. AB - Nosema locustae is a microsporidian parasite of grasshopper pests that is used as a biological control agent, and is one of the emerging model systems for microsporidia. Due largely to its diplokaryotic nuclei, N. locustae has been classified in the genus Nosema, a large genus with members that infect a wide variety of insects. However, some molecular studies have cast doubt on the validity of certain Nosema species, and on the taxonomic position of N. locustae. To clarify the affinities of this important insect parasite we sequenced part of the rRNA operon of N. locustae and conducted a phylogenetic analysis using the complete small subunit rRNA gene. Nosema locustae is only distantly related to the nominotypic N. bombycis, and is instead closely related to Antonospora scoticae, a recently described parasite of bees. We examined the ultrastructure of mature N. locustae spores, and found the spore wall to differ from true Nosema species in having a multi-layered exospore resembling that of Antonospora (one of the distinguishing features of that genus). Based on both molecular and morphological evidence, therefore, we propose transferring N. locustae to the genus Antonospora, as Antonospora locustae n. comb. PMID- 15134258 TI - Nematopsis gigas n. sp. (Apicomplexa), a parasite of Nerita ascencionis (Gastropoda, Neritidae) from Brazil. AB - A new species of Nematopsis (Apicomplexa, Porosporidae) is described from the mantle tissues of the seawater gastropod, Nerita ascencionis (Neritidae), collected in the Atlantic North off the coast of "Fernando de Noronha" Island (3 degrees 47' 57'' S, 32 degrees 25' 12'' W) situated about 350 km from the northeast coast of Brazil. Numerous oocysts, each contained in a parasitophorous vacuole, were found in the cytoplasm of phagocytes in the mantle tissue of the host. The phagocytes were surrounded by a thin wall composed of lucent material. The phagocyte cytoplasm contained a nucleus surrounded by numerous vesicles and some dense masses. The oocysts were 21.9 +/- 0.5 microm long, and 11.5 +/- 0.6 microm wide. The oocyst wall was 0.18-0.25 microm thick, and the apical zone contained a micropyle, 1.0-1.2 microm in diameter, covered by a canopy-like operculum about 0.25 microm thick. Externally, the oocyst wall was surrounded by numerous anastomosing microfibrils attached to the wall and extending towards the periphery of the parasitophorous vacuole. Some microfibrils formed a dense complex network that surrounded the oocyst in the middle of the parasitophorous vacuole, which opened only at the apical zone near the external region of the opercular system. On the basis of the data obtained by light and transmission electron microscopy and host specificity, the gregarine Nematopsis gigas is distinguished from the nearest species as a new species. The taxonomic affinities and morphological comparisons with other similar species of the same genus are discussed. PMID- 15134259 TI - The ultrastructure of the cyst wall of Giardia lamblia. AB - Giardiasis is the most common human protozoal infection. In their cystic phase, giardias are protected from the environment by a filamentous cyst wall made up of carbohydrates, proteins, and by two outer membranes separated from the plasma membrane of the parasite by a peripheral space. The present transmission electron microscope observations of G. lamblia cysts of human origin suggest that the extracellular peritrophic space originates from the growth, elongation, and fusion of large cytoplasmic vacuoles. As the large clear vacuoles grew in size, flattening against the inner face of the plasma membrane, they formed a single vacuole that surrounded the body of the parasite, eventually forming two outer membranes. In mature Giardia cysts, the original plasma membrane of the trophozoite becomes the outermost membrane of the cyst wall (CM1). The large vacuoles form a second membrane surrounding the cyst (CM2), and also form a third membrane (CM3), that becomes the new plasma membrane of the trophozoite. During excystation CM1 and CM2 attach to each other and fragment, leaving abundant membrane residues in the peritrophic space. Knowledge of the biochemical composition and functional properties of the complex outer membranous system of G. lamblia cysts here described will be of use to understand the survival of Giardia cysts in the environment, a major factor responsible for the high prevalence of giardiasis worldwide. PMID- 15134260 TI - Highly divergent actins from karyorelictean, heterotrich, and litostome ciliates. AB - We have cloned, sequenced, and characterized cDNA of actins from five ciliate species of three different classes of the phylum Ciliophora: Karyorelictea (Loxodes striatus), Heterotrichea (Blepharisma japonicum, Blepharisma musculus), and Litostomatea (Didinium nasutum, Dileptus margaritifer). Loxodes striatus uses UGA as the stop codon and has numerous in-frame UAA and UAG, which are translated into glutamine. The other four species use UAA as the stop codon and have no in frame UAG nor UGA. The putative amino acid sequences of the newly determined actin genes were found to be highly divergent as expected from previous findings of other ciliate actins. These sequences were also highly divergent from other ciliate actins, indicating that actin genes are highly diverse even within the phylum Ciliophora. Phylogenetic analysis showed high evolutionary rate of ciliate actins. Our results suggest that the evolutionary rate was accelerated because of the differences in molecular interactions. PMID- 15134261 TI - Identification of a second rRNA gene unit in the Perkinsus andrewsi genome. AB - Perkinsus species are parasitic protozoa of mollusks, currently classified within the Perkinsozoa, a recently established phylum that is basal to the Apicomplexa and Dinozoa. Ribosomal RNA (rRNA) genes and their intergenic spacers have been used to support the taxonomy of Perkinsus species, the description of new species, and to develop molecular probes for their detection and identification. We previously described ultrastructure, behavior in culture, and partial sequence of the rRNA locus of a Perkinsus species isolated from the baltic clam Macoma balthica. The rRNA genes and intergenic spacers of this Perkinsus isolate differed from those described in the currently accepted species to a degree that led to its designation as a new species, Perkinsus andrewsi. In this study, we identify an additional rRNA gene unit (rRNA-B) in the P. andrewsi holotype, and report the complete sequences of both rRNA gene units. Except for the 5.8S, all regions of the rRNA-B gene unit exhibited sequence differences from that initially described (rRNA-A). Each rRNA gene unit is arranged in a "head-to-tail" tandem repeat. This is the first report demonstrating two distinct rRNA units in a Perkinsus species. PMID- 15134262 TI - Ellobiopsids of the genus Thalassomyces are alveolates. AB - Ellobiopsids are multinucleate protist parasites of aquatic crustaceans that possess a nutrient absorbing 'root' inside the host and reproductive structures that protrude through the carapace. Ellobiopsids have variously been affiliated with fungi, 'colorless algae', and dinoflagellates, although no morphological character has been identified that definitively allies them with any particular eukaryotic lineage. The arrangement of the trailing and circumferential flagella of the rarely observed bi-flagellated 'zoospore' is reminiscent of dinoflagellate flagellation, but a well-organized 'dinokaryotic nucleus' has never been observed. Using small subunit ribosomal RNA gene sequences from two species of Thalassomyces, phylogenetic analyses robustly place these ellobiopsid species among the alveolates (ciliates, apicomplexans, dinoflagellates and relatives) though without a clear affiliation to any established alveolate lineage. Our trees demonstrate that Thalassomyces fall within a dinoflagellate + apicomplexa + Perkinsidae + "marine alveolate group 1" clade, clustering most closely with dinoflagellates. However, the poor statistical support for branches within this region indicates that additional data will be needed to resolve relationships among these taxa. PMID- 15134264 TI - More bad news about medical litigation. PMID- 15134265 TI - Lung cancer guidelines: why do we need them? PMID- 15134263 TI - Localization by indirect immunofluorescence of tetrin, actin, and centrin to the oral apparatus and buccal cavity of the macrostomal form of Tetrahymena vorax. AB - We have taken advantage of the size of the macrostomal oral apparatus of Tetrahymena vorax to investigate the immunofluorescent localization of three cytoskeletal proteins--tetrin, actin, and centrin. Tetrin and actin antibodies co localize to cross-connectives that anchor the membranelles. These antibodies also recognize the coarse filamentous reticulum, a filament associated with the undulating membrane. Actin-specific localization extends beyond the coarse filamentous reticulum-undulating membrane complex into a region called the specialized cytoplasm. A centrin antibody localizes to the fine filamentous reticulum which, along with microtubules of the oral ribs, circumscribes the cytostomal opening. Models of phagocytic contraction based on these data are presented. PMID- 15134266 TI - The role of surgery in the management of obstructive sleep apnoea syndrome in adults. PMID- 15134267 TI - An evaluation of left sided colonic polyps as a marker for right sided cancers. AB - The detection of a left-sided polyp on flexible sigmoidoscopy has been suggested as providing a heralding sign for right-sided tumours. We assessed our own experience with right-sided colonic cancers with reference to detection modality, surgical intervention and their association with left-sided polyps. We performed a retrospective review of a prospectively collected colorectal cancer database, with endoscopic and radiological records. Patients were subcategorised on the basis of the presence or absence of left-side polyps. Ninety-one consecutive patients who underwent curative surgery for right-sided colonic cancers were studied. Endoscopy was used to detect right-sided carcinomas in 10 (83%) of the 12 cases with synchronous left-sided polyps and radiological imaging utilised in 2 cases. In patients without evidence of left sided disease endoscopy was used in 40 (51%) of the 79 patients. In our experience most right sided cancers do not have a synchronous polyp evident on the left side 79 (87%). PMID- 15134268 TI - Pertussis--going, going, but not gone: thirteen year trends in the incidence of pertussis in the Republic of Ireland and North Eastern Health Board. AB - Pertussis or whooping cough is caused by Bordetella pertussis. Complications of pertussis include pneumonia, seizures, encephalopathy and death. Diagnostic tests include pernasal swab culture, serology, direct immuno-fluorescent assay (DFA) and polymerase chain reaction (PCR). At present, the majority of cases are diagnosed clinically. DTaP immunisation uptake has considerably reduced disease incidence. We describe national and NEHB reported cases of pertussis over a 13 year period from 1988 to 2000 inclusive and we also describe in detail all pertussis admissions to Our Lady of Lourdes Hospital, Drogheda from 1998 to 2000 inclusive. From 1988 to 2000 inclusive, all pertussis cases notified nationally and in the NEHB region were tabulated and studied from data obtained from the Department of Public Health, NEHB and the NDSC, together with national and NEHB DTaP immunisation rates. All admissions to Our Lady of Lourdes Hospital, Drogheda with a diagnosis of pertussis from 1998 to 2000 inclusive were studied in detail by retrospective medical record review and the results were later analysed using the Epi-Info Version 6.0 statistical package. National notified pertussis cases had fallen from 1170 cases in 1988 to 150 cases in 2000. There was a striking rise in pertussis nationally in 1989 to 2217 cases and this directly related to the sharp fall in DTaP immunisation uptake at that time due to adverse publicity. Current 5-in-1 immunisation rates are between 80% to 85% nationally, and 88% for the NEHB region. NEHB pertussis notifications rose from 72 cases in 1988 to 328 in 1989 and this directly reflected the national trend; in 2000 there were only 13 notifications. In the NEHB region between 1998 and 2000 inclusive, there were only 26 notified cases of pertussis, 17 of which were admitted to hospital. Only 1/17 (6%) had a positive pernasal swab culture. There were no deaths. Pertussis is under-diagnosed and is still largely a clinical diagnosis. There has been an approximate ten-fold decrease in pertussis incidence rates over the 13 year study period both nationally and in the NEHB region. Current DTaP immunisation rates in the NEHB region of 88%, and of between 80% to 85% nationally are insufficient to confer 'herd immunity' and thus young infants will continue to be at risk. PMID- 15134269 TI - Needs analysis for tele-neurophysiology in the Irish North-Western Health Board. AB - No clinical neurophysiology (CN) service is currently provided by the North Western Health Board (NWHB) region in Ireland. Digital technology associated with CN is compatible with tele-neurophysiology service development and may increase the efficiency of patient care in remote areas. This study was conducted to assess the need for CN in the NWHB with a view to implementing a tele neurophysiology service for the region. A retrospective audit of investigations performed at six CN departments in Dublin compared patient groups from regions with (Eastern Regional Health Authority--ERHA) and without (NWHB) local CN departments. 4954 records were audited. CN activity was lower than internationally expected (ERHA group 52% of expected, NWHB group 23% of expected). The 2 groups differ in the specialty of referring clinician (p < 0.0001). NWHB patients are more likely to be referred by clinicians outside their region (p < 0.0001) and wait longer for appointment (p < 0.0001). While there is an unmet demand for CN services in Ireland generally, the population of the NWHB is further disadvantaged. Tele-neurophysiology has the potential to provide fairer access to CN services across the country. The data from this study provide a baseline against which the outcome of investment in CN can be monitored and evaluated. PMID- 15134270 TI - Abdominal ultrasound in the evaluation of the asymptomatic patient with abnormal liver function tests. AB - We retrospectively evaluated abdominal ultrasound studies of 273 asymptomatic patients with abnormal liver function tests over an 18-month period to assess the value of abdominal ultrasound as an initial investigation. In-patients, out patients and general practitioner referrals were included. 143 (53%) examinations were normal. Those with significant abnormality (n=13, 5%) were referred for further investigation/treatment. Those with alterations in hepatic echogenicity (n=117, 42%) underwent clinical and biochemical follow-up, radiological follow-up or biopsy. 20% of patients underwent further radiological investigation. Ultrasound is a non-invasive, cost effective first-line investigation in the evaluation of asymptomatic patients with abnormal liver function tests. PMID- 15134271 TI - Wegener's granulomatosis masquerading as unilateral sinusitis. AB - Wegener's Granulomatosis (WG) is a rare, multi-system disease of unknown aetiology. The disease is characterised by necrotizing granulomas and vasculitis of the upper and lower respiratory tracts and kidneys. The sinonasal region may be the first area to manifest this systemic condition. If the disease is diagnosed early and treated appropriately, involvement of the lungs and kidneys may be averted. Left untreated, the disease may prove rapidly fatal. A low threshold of clinical suspicion must be maintained. We report a case, of a middle aged patient who presented with unilateral sinusitis secondary to a large obstructive nasal mass. Anti neutrophil cytoplasmic antibody (ANCA) titres helped confirm the diagnosis of WG. Unilateral sinusitis with a nasal mass, is a very unusual presentation of WG, and emphasises the importance of a low threshold of suspicion for patients presenting with atypical sinonasal symptoms. PMID- 15134272 TI - Massive haemoptysis: causes and management. PMID- 15134273 TI - A survey of parental knowledge of paediatric oral health in Ireland. PMID- 15134274 TI - Safety of home delivery compared with hospital delivery in the Eastern Region Health Authority in Ireland in the years 1999-2002. PMID- 15134275 TI - Dissection of vertebral artery. PMID- 15134276 TI - Cost of teaching stroke in an Irish teaching hospital. PMID- 15134277 TI - High morbidity expected from cirrhosis in injecting drug users. PMID- 15134278 TI - Serotonin receptor ligands and the treatment of obesity. AB - It was first established in the 1970s that the brain serotonin (5-HT) system was involved in the control of eating. Subsequent progress in the molecular pharmacology of 5-HT receptors, and the development of selective 5-HT receptor ligands, has clarified our understanding of the role of 5-HT in the regulation of ingestive behavior. Of the 14 5-HT receptor subtypes currently described, 5-HT1A, 5-HT1B and 5-HT2C receptors have been of principal interest in the regulation of food intake. This is largely due to the development of suitable agonists, antagonists and gene-knockout animals with which the role of these receptors can be elucidated. The recent development of selective ligands and knockout mice for other 5-HT receptors, including the 5-HT2B and 5-HT6 receptors, has also suggested a role for these receptor subtypes in eating behavior. Studies using such approaches should further our understanding of the role of serotonin in the regulation of feeding behavior and thus, may lead to the development of novel, safe, serotonin receptor ligands for the treatment of obesity. PMID- 15134279 TI - Cannabinoid receptor antagonists and obesity. AB - The cannabinoid-1 (CB1) receptor plays a role in the regulation of appetitive behavior. Exogenously administered cannabinoid receptor agonists stimulate food consumption in animals and humans. Endogenous cannabinoid receptor agonists are present in the brain, and the brain level of these agonists increases with greater demand of food by rodents. Specific CB1 receptor antagonist compounds have been discovered that display high affinity and selectivity for the CB1 receptor. CB1 receptor antagonists inhibit both acute and long-term food intake in rodents. Chronic treatment with CB1 antagonists results in a sustained reduction in body weight in rodents (5 weeks), and weight loss in humans (16 weeks). Patent literature indicates CB1 receptor antagonist discovery efforts at a number of pharmaceutical companies. The CB1 receptor antagonist, rimonabant (SR 141716), discovered by Sanofi-Synthelabo, is in phase III clinical trials for the treatment of obesity and has been found to decrease appetite and body weight in humans. PMID- 15134280 TI - Interdependence of hepatic lipid and glucose metabolism: novel pharmacological targets for diabetes. AB - Type 2 diabetes is a growing world epidemic which is fuelled by the increased prevalence of obesity, together with changes in lifestyle and nutritional habits. Current medical treatments for type 2 diabetes include stimulation of insulin secretion, inhibition of endogenous glucose production and enhancement of insulin sensitivity. However, more efficacious therapies are needed. Aberrations in the control of energy metabolism and substrate utilization are one of the hallmarks of insulin resistance and type 2 diabetes, and is reflected by tendencies of excessive accumulation of lipid products in tissues such as skeletal muscle, liver and the pancreatic islet beta-cell. Recent developments in understanding the interaction between carbohydrate and fatty acid metabolism may guide us to novel pharmacological targets in this area. PMID- 15134281 TI - The potential role of glucagon-like peptide 1 in diabetes. AB - The incretin hormone glucagon-like peptide 1 (GLP-1) has a promising potential for the treatment of type 2 diabetes due to its glucose-dependent insulinotropic and glucagonostatic properties. In addition, the peptide potently decelerates gastric emptying and inhibits appetite, thereby leading to reduced food intake. In animal studies, GLP-1 has been demonstrated to increase B-cell mass via inhibition of apoptosis and stimulation of B-cell replication and neogenesis. However, an in vivo half-life in the range of minutes limits the therapeutic use of the native peptide GLP-1. Different pharmacological approaches to overcome these problems are currently being evaluated. They include the continuous parenteral administration of the peptide via infusion pumps, the inhibition of its in vivo degradation and the generation or use of modified derivatives/analogs of GLP-1 displaying prolonged biological activity. The physiological effects of GLP-1 and its pharmacokinetic limitations will be reviewed here, and the current therapeutic approaches based on GLP-1 discussed. PMID- 15134282 TI - Inhibition of glycerol-3-phosphate acyltransferase as a potential treatment for insulin resistance and type 2 diabetes. AB - This review focuses on the potential of glycerol-3-phosphate acyltransferase (GPAT) inhibition as a strategy to treat insulin resistance, one of the characteristics of obesity and type 2 diabetes. Inhibition of GPAT, which catalyzes the first and committed step in triacylglyceride synthesis, has the potential to reduce accumulation of ectopic fat in insulin-sensitive organs such as the liver and skeletal muscle. Such an accumulation of fat has been shown to be correlated with insulin resistance. Thus, its reduction by pharmacological treatment is an attractive strategy to treat type 2 diabetes. Potential methods to identify inhibitors for acyltransferases suitable for treatment of human diseases are described. PMID- 15134283 TI - Growth factors for skeletal reconstruction and fracture repair. AB - The demographic challenges of an ageing population have emphasized the need for processes to augment and repair skeletal tissue loss as a consequence of trauma and/or degeneration. A number of bone growth factors have been shown to be expressed during the course of fracture healing, suggesting a potential role in bone and cartilage formation, and in fracture repair. This review focuses on a select number of these growth factors currently under preclinical and clinical evaluation for skeletal regeneration and fracture repair. The limitations in the use of these skeletal factors to augment bone growth, thus improving quality-of life and reducing the significant social and economic costs associated with skeletal trauma/loss are also considered. PMID- 15134284 TI - Tocolysis: current controversies, future directions. AB - The use of tocolytic agents for the treatment of preterm labor is not supported by the current evidence from placebo-controlled trials. Despite this, tocolytics continue to be widely used in clinical practice. Possible reasons for the lack of observed benefit include the hypothesis that suppression of labor may be harmful in some situations, along with a failure to appreciate the implications of the complex physiology of parturition. Importantly, however, it must be recognized that the quality of evidence addressing the question is poor, with studies significantly underpowered to detect changes in health outcomes. Where tocolysis is used, recent trends have favored agents with lower maternal side effect profiles, including calcium channel blockers and the oxytocin receptor antagonist atosiban. The use of cyclooxygenase-selective inhibitors of prostaglandin synthesis is currently being explored. Future directions in tocolytic research include the use of multiple agent therapies, along with the development of more selective treatments with low side effect profiles. Such agents include oxytocin receptor antagonists with more favorable pharmacological properties and prostaglandin F2alpha receptor antagonists. Future research into tocolytic therapies must focus on evaluating health outcomes from treatments rather than simply the ability to prolong pregnancy, and consequently the design of appropriate clinical studies needs careful consideration with respect to issues such as inclusion criteria, sample size and the selection of appropriate outcome measures. PMID- 15134285 TI - Rimonabant Sanofi-Synthelabo. AB - Rimonabant, an antagonist of central cannabinoid type 1 (CB1) receptors, is being developed by Sanofi-Synthelabo for the potential treatment of obesity and as a potential smoking cessation agent. Phase III trials were initiated for obesity in August 2001 and were ongoing in September 2003. By September 2002, the compound had entered phase III trials for smoking cessation, and these trials were ongoing in September 2003. PMID- 15134286 TI - AOD-9604 Metabolic. AB - Metabolic is developing AOD-9604 for the potential treatment of obesity. By February 2002, phase IIa trials were underway. PMID- 15134288 TI - Ospemifene Hormos. AB - Hormos Medical is developing the selective estrogen receptor modulator ospemifene, for the potential treatment of postmenopausal osteoporosis and urogenital atrophy. By November 2003, Hormos had advanced the drug into phase II/III testing for both indications. PMID- 15134287 TI - ED-71 Chugai. AB - Chugai is developing ED-71, an orally available vitamin D3 derivative, for the potential treatment of osteoporosis. By December 2003, the compound had entered phase III trial for the prevention of fracture. PMID- 15134289 TI - PT-141 Palatin. AB - Palatin, under license from Competitive Technology, is developing the peptide PT 141, a synthetically modified analog of PT-14, as a nasal spray for the potential treatment of erectile dysfunction (ED) and female sexual dysfunction. In September 2003, Palatin had completed a phase IIb trial in patients with ED and in February 2004, Palatin planned to start phase III trials in early 2005. PMID- 15134290 TI - Pegvisomant Pfizer/Sensus. AB - Pfizer (formerly Pharmacia), in collaboration with its wholly owned subsidiary Sensus, has developed and launched pegvisomant, a pegylated, genetically modified human growth hormone (hGH), for the treatment of acromegaly. Pegvisomant, in contrast to classical somatostatin analogs which lower hGH synthesis, exerts its anti-hGH action by preventing GH receptor activation. This drug is now available in the US and Europe for the treatment of acromegaly. PMID- 15134291 TI - Fluoroquinolone-resistant Campylobacter in animal reservoirs: dynamics of development, resistance mechanisms and ecological fitness. AB - Thermophilic Campylobacter species, including Campylobacter jejuni and Campylobacter coli, are responsible for foodborne campylobacteriosis in humans and are increasingly resistant to fluoroquinolone (FQ) antimicrobials. The therapeutic use of FQ antimicrobial agents in food animal production, particularly in poultry, has become a concern for public health, because the practice may promote the emergence of FQ-resistant Campylobacter that can be transmitted to humans through the food chain. Recent studies have indicated that Campylobacter displays a hypermutable phenotype in response to in vivo treatment with FQ antimicrobials, resulting in the rapid emergence of resistant mutants. Distinct from other Gram-negative bacteria, the acquisition of FQ resistance in Campylobacter does not require stepwise accumulation of gyrA mutations and overexpression of efflux pumps, and is mainly mediated by single-step point mutations in gyrA in the presence of a constitutively expressed multidrug efflux pump, CmeABC. The simplicity of the resistance mechanisms may facilitate the rapid adaptation of Campylobacter to FQ treatment. The FQ-resistant Campylobacter mutants derived from chickens do not show a fitness cost in vivo and are ecologically competitive in the colonization of chickens even in the absence of antimicrobial selection pressure. These findings suggest that FQ-resistant Campylobacter may continue to persist regardless of antimicrobial usage, and highlight the need for extra effort to prevent the occurrence and spread of FQ resistant Campylobacter in animal reservoirs. PMID- 15134292 TI - Molecular genetic methods in the veterinary clinical bacteriology laboratory: current usage and future applications. AB - In the last 5 years, numerous molecular methods have been published for the detection and characterization of bacteria in the field of veterinary medicine. PCR has been the most commonly used technology. Although not currently used for clinical veterinary diagnosis, new technologies such as liquid-phase hybridization, real-time PCR, pathogen load determination and DNA/protein microarray have been described and have many possible applications in the clinical bacteriology laboratory because of their sensitivity and efficiency. This review describes the basic principles and application of recently published DNA-based molecular techniques for the purpose of veterinary clinical bacteriological diagnosis. It covers advances in probe hybridization technology, DNA/RNA amplification techniques and other molecular detection methods, including 16S rRNA analysis for bacterial characterization and DNA microarrays for bacterial detection. The review briefly summarizes the application of molecular methods for the diagnosis of specific important bacterial infections of animals, and for other animal pathogens that are slow or difficult to isolate in the clinical bacteriology laboratory. In addition, the molecular detection of antimicrobial resistance genes and of bovine mastitis pathogens is briefly described and current commercially available tests are listed. PMID- 15134293 TI - Cryptosporidium and its potential as a food-borne pathogen. AB - Cryptosporidium species are intestinal protozoan parasites and are excreted in animal feces as stable oocysts. Cryptosporidium has now been detected in the feces of a wide range of ruminant and non-ruminant farmed animals, wild animals, domestic pets and birds and the parasite appears to be well adapted to survive and persist in feces for extended periods, ranging from several weeks to many months. Because of this persistence, these materials are important as potential vehicles of transmission within herds, farms, the water chain, the fresh food chain, and the wider environment. Appropriate handling of animal waste is necessary to control spread of this pathogen and to limit the significant risks of human infection. While water is a well-recognized vector of Cryptosporidium, it has only recently emerged that food may play a more significant role than previously realized in the transmission of the Cryptosporidium to humans. In the last 3-5 years, research efforts have been directed both at the development of suitable methods for isolation and detection of the parasite in foods and at the application of these methods to assess the prevalence and persistence of the parasite in a range of foods. Additionally, molecular subtyping methods have been used to establish the transmission routes of the parasite. This paper summarizes the general biology of Cryptosporidium and overviews the current research on C. parvum in the food chain. The risks posed by certain foods, such as salad/vegetable crops and beef, are discussed and control measures which may be useful in the farm-to-fork chain for these products are described. PMID- 15134294 TI - Equine cardiovascular pathology: an overview. AB - The few data collections that evaluate the involvement of organ systems in horse diseases are in agreement that the locomotor, gastrointestinal and nervous systems are the sites of primary disease in the vast majority of sick horses. When compared with diseases of these organ systems, equine cardiovascular diseases occur infrequently. The most detailed and comprehensive survey of equine cardiac pathology was reported in 1972 by Else and Holmes, who summarized the gross and microscopic cardiac findings from 1500 abattoir horses. This paper reviews the pathology of the cardiovascular diseases typically encountered in horses. Most of the pathological examples are from the files of the Veterinary Medical Teaching Hospital at the University of Florida, encompassing 24 years (1978-2002) and some 6000 equine necropsy cases. Preceding the specific topics are principles of the anatomy and function of the normal equine heart. Pathological entities include equine congenital cardiovascular diseases, acquired diseases of the pericardium, myocardium, endocardium and valves, cardiac neoplasms, and common equine vascular diseases and vascular neoplasms. Extensive use is made of photographs to illustrate the features of individual case examples. PMID- 15134295 TI - The biology of dendritic cells and their potential use in veterinary medicine. AB - Dendritic cells have been shown to be the main antigen-presenting cells in vitro and in vivo, playing a pivotal role in the induction of both tolerance and immunity. Dendritic cells from humans and mice have been extensively studied and dendritic cell-based vaccines have been shown to be effective in the prevention and treatment of infectious, allergic and neoplastic diseases. Studies of dendritic cells of domestic animal origin are becoming available and confirm a role for these cells in the pathogenesis of a variety of animal diseases, suggesting that dendritic cells could be used as adjuvants for prophylactic and therapeutic strategies in veterinary medicine. PMID- 15134296 TI - A review of porcine tonsils in immunity and disease. AB - The porcine tonsils are a group of lymphoepithelial tissues located at the common openings of the gastrointestinal and respiratory tracts. The tonsils participate in a variety of functions involving innate, cellular and humoral immunity at the local and systemic levels. Among these immunological functions is the continuous surveillance for the presence of foreign antigens at the openings of the gastrointestinal and respiratory tracts. Within the pig, the movement of lymphocytes, cytokines and chemotactic molecules from the tonsils to other lymphoid organs confers immunity to other portals of pathogen entry and facilitates an efficient and rapid systemic immune response. In spite of the immunological nature of the tonsils, some microorganisms have acquired adaptations that allow them to circumvent the tonsillar immune defenses and utilize the tonsils as a site of entry, replication and colonization. Several bacterial and viral pathogens persist asymptomatically within the tonsils, making identification of asymptomatic carrier animals difficult in disease control and/or pathogen elimination. This paper reviews the current information on the anatomy, immunology and pathobiology of porcine tonsils and discusses the tonsils as a site of pathogen entry, replication and colonization using Salmonella spp., classical swine fever virus and porcine reproductive and respiratory syndrome virus as examples. PMID- 15134297 TI - Zoonotic skin diseases of dogs and cats. AB - Although there are over 250 zoonotic diseases, only 30-40 of them involve dogs and cats. Transmission of zoonotic infections occurs via bites, scratches or touch; exposure to saliva, urine or feces; inhalation of particles or infectious aerosols; contact with a transport or intermediate host (e.g. ticks, fleas); or exposure to contaminated water, soil or vegetation. This paper summarizes the most important common zoonotic dermatological diseases of dogs and cats. The most common dermatological zoonoses are flea and tick infestations and the diseases they transmit; dermatophytosis; and mite infestations (Sarcoptes and Cheyletiella). Prevention of zoonotic infestations or infections can be accomplished easily by the use of routine flea and tick control, screening of new pets for dermatophytosis, and routine hand-washing. PMID- 15134298 TI - The future of pediatric endocrinology. PMID- 15134299 TI - Genetics of growth retardation. AB - Genetic disorders of growth are numerous and include defects of the central nervous system, hypothalamus, pituitary, thyroid, skeleton and other glands, tissues or organs. These disorders are caused by genetic variations that interrupt different points of the hypothalamic-pituitary axis and other synthetic pathways that are required for growth. The resulting growth disorders can be classified by their 1) etiology (Mendelian, mode of inheritance or chromosomal basis), 2) phenotype (isolated or multiple hormone deficiencies), 3) genetic variations of different loci or alleles involved, and/or 4) the associated biochemical findings. The growth disorders selected for inclusion here are examples and are by no means a comprehensive list of all known genetic diseases that have some effect on growth. Information on each disorder is summarized and, for most, more detailed information can be obtained through the electronic database Online Mendelian Inheritance in Man (OMIM) which is available at http://www.ncbi.nlm.nih.gov/Omim. PMID- 15134300 TI - Therapeutic optimization of growth hormone deficiency in children and adolescents. AB - More than 40 years after the introduction of growth hormone (GH) treatment, many questions remain unanswered. Clearly, with the availability of rhGH and with current treatment protocols, treatment efficacy has improved. However, it still remains unclear whether current treatment protocols are the best possible. Before GH deficiency was recognized as a chronic disease, children only received treatment until normal adult height had been reached. However, it has recently been shown that not all GH-dependent body structures and functions normalize in parallel with height. Furthermore, in adolescents with GH deficiency, the interruption of GH substitution leads to severe hormone deficiency symptoms in adulthood. In the case of an adolescent who meets the biochemical criteria for GH deficiency in adulthood, but does not show alterations of metabolism, body structure, or emotional state, should GH treatment be started in adolescence, or only if and when the clinical syndrome becomes apparent? This is a difficult question to which there is not yet any clear answer, and we suggest that there is a need for further studies in this area. Furthermore, it will be necessary to re evaluate the situation of patients who have completed their growth, and definitive conclusions will require controlled studies. PMID- 15134301 TI - Childhood-onset congenital adrenal hyperplasia: long-term outcome and optimization of therapy. AB - Congenital adrenal hyperplasia is a general term applied to several disorders caused by inherited recessive defects of cortisol synthesis. The most common form is 21-hydroxylase deficiency, accounting for 95% of cases. The classical forms have an incidence of one in 15,000 and the non-classical forms about one in 1,000. The classical or severe phenotype presents in the newborn period or early infancy with virilization and adrenal insufficiency, with or without salt-losing; the non-classical or mild phenotype presents in late childhood or early adulthood with signs of hyperandrogenism. This wide range of clinical expression is explained by genetic variation. Although there is a certain amount of genotype phenotype correlation, discrepancies have been described. During the last 30 years there has been a substantial improvement in diagnosis and treatment of this disease, and patients with CAH now reach adulthood. Treatment of this condition is intended to reduce excessive corticotropin secretion and replace glucocorticoids and mineralocorticoids as physiologically as possible. Clinical management is often complicated by periods of inadequately treated hyperandrogenism, iatrogenic hypercortisolism, or both. Long-term consequences in adult life may include short stature, obesity, diminished bone mass, gonadal dysfunction with low fertility rates and psychosexual dysfunction in females. New treatment approaches are under investigation, such as the use of anti-androgens, inhibitors of estrogen production and adrenalectomy for severely resistant cases. PMID- 15134302 TI - Implications for adulthood and therapeutic optimization in diabetes mellitus. PMID- 15134303 TI - Optimization of treatment in Turner's syndrome. AB - We discuss treatment for short stature and hypogonadism in patients with Turner's syndrome, using recombinant human growth hormone (GH) and estrogen. We discuss the starting dose of GH, age at start of treatment, and association with oxandrolone or low dose estrogen therapy before 12 years old. We provide recommendations for optimization of therapy for best final height and induction of puberty with estrogen therapy. Our experience in the Children's Endocrinology Clinic in "La Paz" University Hospital, Madrid, is that 83% of our patients need estrogen therapy and only 17% had spontaneous menarche. The average age for puberty induction was 12.2 years, and the average duration of estrogen treatment was 2.1 years. The average final height obtained using GH treatment was 151.3 cm with a gain related to predicted height of 6.8 cm. PMID- 15134304 TI - Fetal growth regulation and intrauterine growth retardation. AB - Gestational age and neonatal anthropometric parameters are currently used to evaluate fetal growth and are predictive factors of perinatal and postnatal morbidity and mortality. We performed a retrospective analysis of neonatal anthropometric parameters (weight, vertex-heel length and head circumference) in 1,470 live preterm neonates born between 1997 and 2002 and a prospective analysis of the same parameters in 1,786 live newborns of both sexes born in 2001 and 2002, products of single 37-42 week uncomplicated pregnancies in healthy Spanish Caucasian mothers. A progressive increase in these parameters with gestational age and sexual dimorphism were observed from the 30th week of gestational age onwards, with statistically-significant differences (p<0.05) at 38-42 weeks of gestational age. An increase in weight and length values in relation to previous Spanish studies was also documented in preterm newborns. It is estimated that 10 15% of children born small for gestational age (SGA) do not experience catch-up growth by the age of 3 years and may have short stature in adulthood. Preliminary data of a cross-sectional study on spontaneous growth in boys and girls born SGA without postnatal catch-up growth show that their +2 SD values of height are similar to -2 SD values of our normal control population of children born with adequate weight and length for gestational age (AGE). However, weight +2 SD values are similar to mean values of control children born AGE. In summary, our data show sexual dimorphism in neonatal anthropometric growth parameters and that these parameters change with time and may be updated. In addition children born SGA without postnatal catch-up are shorter and have higher weight than age-, height- and sex-matched controls born AGE. PMID- 15134305 TI - Study of genetic expression of intrauterine growth factors IGF-I and EGFR in placental tissue from pregnancies with intrauterine growth retardation. AB - To investigate the possibility of altered gene expression of growth factors in prenatal growth retardation, we assessed expression of the genes for insulin-like growth factor-I (IGF-I) and epidermal growth factor receptor (EGFR) by RT-PCR from human placentas at term delivery in two groups: appropriate for gestational age (AGA) and pregnancies complicated with IUGR. The placentas from IUGR gestations showed reduced IGF-I expression with a significance of p = 0.008, whereas we did not find any significant differences in EGFR gene expression. PMID- 15134306 TI - Intrauterine growth retardation: study of placental apoptosis. AB - We studied the rate of apoptosis in the placental tissue of pregnancies complicated with intrauterine growth retardation (IUGR) and compared it with the results obtained in normal placentas. Our results clearly demonstrate a strongly increased rate of apoptosis in placentas of children born with IUGR, suggesting severe placental dysfunction. The significance of these findings needs further study. PMID- 15134307 TI - Psychomotor and intellectual development of children born with intrauterine growth retardation (IUGR). AB - The possible impact of IUGR on the intellectual outcome of children born with IUGR gives special relevance to this condition. In order to determine the psychomotor and intellectual development of such children, we analyzed the evolution of 60 children through appropriate tests, along the years, and the possible influence of two factors, the socio-economic status of the family, and whether or not there was catch-up growth. Our results show a negative impact of IUGR on the intellectual outcome of these children, independent of catch-up growth, although those with catch-up growth showed better evolution. The socio economic status plays a limited role only at older age. Those children followed longitudinally for 1 year did not show any amelioration of their IQ. PMID- 15134308 TI - Small for gestational age (SGA): endocrine and metabolic consequences and effects of growth hormone treatment. AB - Several studies have demonstrated an association between low birth weight and impaired insulin sensitivity or even type 2 diabetes mellitus (DM2) in later life. Growth hormone (GH) is known to increase fasting and postprandial insulin levels. For that reason concern has been expressed regarding possible detrimental effects of GH therapy in children born SGA. In a Dutch trial the possible side effects of GH therapy on carbohydrate metabolism were assessed in short children born SGA after 6 years and at 6 months after discontinuation of GH therapy. This study included 79 prepubertal short children born SGA, participating in a multicenter double-blind, randomized, dose-response GH trial. Inclusion criteria were: 1) birth length SDS below -1.88, 2) age 3-11 years in boys and 3-9 years in girls, 3) height SDS < -1.88, 4) no spontaneous catch-up growth, and 5) an uncomplicated neonatal period. Mean (SD) value for age was 7.3 (2.1) years, birth length SDS -3.6, height SDS -3.0 (0.7) and BMI SDS -1.2 (1.3). All children were randomly assigned to either group A (n = 41) using 1 mg GH/m2/day or group B (n = 38) using 2 mg/m2/ d/ay (approximately 0.1 or 0.2 IU/kg/d, respectively). Standard oral glucose tolerance tests (OGTTs) were performed before and during 6 years of GH therapy and 6 months after discontinuation of GH therapy. Before GH therapy 8% of the children had impaired glucose tolerance (IGT) according to criteria of the WHO. After 6 years of GH therapy, IGT was found in 4% and after stopping GH in 10%. Mean fasting glucose increased significantly with 0.5 mMol/l after 1 year of GH therapy, without a further increase thereafter. GH therapy induced considerably higher fasting and glucose-stimulated insulin levels. None of the observed changes were different between the GH dosage groups. Children who remained prepubertal had similar glucose and insulin levels compared to children who entered puberty. HbA1c levels were always in the normal range and none of the children developed diabetes mellitus. After discontinuation of GH therapy the mean serum glucose levels remained normal and the mean serum insulin levels decreased significantly, to normal age reference values. Before the start of GH the mean systolic blood pressure was significantly higher compared to age-matched peers, whereas during GH therapy a significant decline in mean systolic blood pressure occurred, which remained similar after discontinuation of GH treatment. In conclusion, continuous, long-term GH therapy in short children born SGA has no adverse effects on glucose levels, even with GH dosages up to 2 mg/m2/day. However, as has been reported in other patient groups, GH induced higher fasting and glucose-stimulated insulin levels, indicating insulin resistance. After discontinuation of GH, serum insulin levels declined to normal age-matched reference levels. Since impaired insulin sensitivity and DM2 have been demonstrated in relatively young patients born SGA, long-term follow-up of children born SGA is advised, also after discontinuation of GH therapy. PMID- 15134309 TI - Adolescence today. PMID- 15134310 TI - New concepts in anorexia nervosa. AB - Anorexia nervosa (AN), a psychosomatic disorder, has serious negative effects on multiple organs and systems of the human body. A large number of endocrine and metabolic anomalies have been described, including amenorrhea/oligomenorrhea, delayed puberty, hypothyroidism, hypercortisolism, and alterations in the growth hormone (GH) axis and bone metabolism. The role of different peptides, including ghrelin, leptin, neuropeptide Y and serotonin, in the regulation of appetite is discussed. In addition, isolated hypogonadotropic hypogonadism that occurs in these patients is analyzed in detail, as well as the abnormalities in the growth hormone axis. Alterations in bone mineral density, bone markers and the degree of osteopenia in these patients, depending on the age at which amenorrhea began and its duration, are also discussed. Finally, our current understanding of the possible benefits of treatment with estrogens and progestogens is also analyzed. PMID- 15134311 TI - The need for specialized adolescence units. AB - Adolescence is the period of life of transition from childhood to adulthood; it includes not only sexual maturation but also physical and psychological maturation. The chronological age when this transition occurs is imprecise and not exactly defined. There are specific diseases that appear during this period of life that deserve specialized medical care. This article deals with the need to create specialized adolescence units with the direct participation of pediatricians. The transition through adolescence into adulthood is complicated by physical, psychological and social issues, and the transfer of adolescent patients to adult departments should be done gradually. There is a need for consensus guidelines about this issue. PMID- 15134312 TI - Child-onset growth hormone deficiency in adulthood. Transfer of patients from pediatric to adult endocrinology units. AB - Treatment with growth hormone (GH) in adult-onset GH deficiency (AO-GHD) reverses its many metabolic alterations, modifying body composition, bone mass, several cardiovascular risk factors, and improving quality of life. In adult patients with a previous diagnosis of child-onset GH deficiency (CO-GHD), the lack of treatment also produces similar alterations, reversed by GH treatment. In patients with multiple pituitary hormone deficiency, the lack of GH is considered definitive, but in isolated GHD, the need for re-evaluation of the deficit is mandatory. The 'gold standard' test is insulin-induced hypoglycemia, after a wash out period, and the criterion for GH therapy should be a GH (polyclonal-RIA assay) response less than 3 ng/ml. The initial recommended GH dose is lover than in children, and the dosage must be adjusted to maintain IGF-I levels in the normal range. We propose that decisions about patient recruitment, assessment, confirmation or reevaluation, information about new perspectives, disadvantages and benefits of GH therapy, and the beginning of treatment should be made in cooperation by pediatric and adult endocrinologists, so the patient receives all information from both medical teams, before being transferred to the adult endocrinology department. PMID- 15134313 TI - How do outcomes in a specified parent training intervention maintain or wane over time? AB - In a randomized prevention trial, 238 recently separated mothers and their young sons were assigned to either Parent Management Training (PMT) or a comparison group. Families were intensively assessed at baseline and at each 6-month interval through 30 months. To understand the effects of PMT, we first evaluated effect sizes among family variables over time. Second, because observed parenting was the target of PMT, we hypothesized a sequential pattern of structured changes within and between individuals. Using constructs with mismatched sources of data, we conducted a set of latent growth mediational analyses to test hypothesized mechanisms explaining change. Effect sizes indicated that parenting changed first within 12 months, followed by changes in boy behaviors and finally changes in maternal depression within 30 months. Unique follow-up findings indicated that intervention effects on reductions in maternal depression were mediated by reductions in boy externalizing; intervention effects on externalizing were mediated by reductions in boy depression. As expected, increases in effective parenting predicted reductions in child behavior problems. PMT effects on internalizing were direct and indirect, partially mediated by parenting practices. Results are discussed from a system's perspective on PMT amplifiers. PMID- 15134314 TI - The impact of schools on juvenile substance initiation and use. AB - We use data from the two rounds of the NLSY97 and the corresponding QED data to examine the effectiveness of school endowments and curricula in targeting juvenile use of tobacco, alcohol, and marijuana. Our results support the notion that schools matter in reducing juvenile involvement in substance use. Higher discretionary dollars per pupil are linked to reduced rates of juvenile initiation and repetitive use rates of cigarettes and marijuana. Additionally, school curricula, as indicated by the implementation of year round classes and some innovative and after-school programs--such as gifted and talented, attendance monitoring, homework hotline, international baccalaureate, extended day, and mentoring, programs, affect both juvenile initiation to tobacco and alcohol use and juvenile repetitive use of tobacco and alcohol. In particular, we find that juvenile initiation to cigarette use is approximately between 2 percentage points and 3 percentage points lower among youths attending schools with gifted and talented and international baccalaureate programs. In addition, juvenile repetitive cigarette use is approximately 54%, 52%, and 48% lower among youths attending schools offering year round classes, international baccalaureate, and twenty-first century programs, respectively. Finally, juvenile initiation to alcohol use and juvenile repetitive use of alcohol are approximately 3% and 20% lower, respectively, among youths in schools offering gifted and talented programs. In sum, while these programs are not implemented to address substance use problems among the student body, we find that the implementation of these programs is often accompanied by a reduction in juvenile initiation and repetitive substance use. PMID- 15134315 TI - Increased parent limits on teen driving: positive effects from a brief intervention administered at the Motor Vehicle Administration. AB - The purpose of this study was to determine whether exposure to a brief intervention administered at the Motor Vehicle Administration (MVA) increases parental limits on teen driving. A total of 658 parents and their 16-year-old adolescents were recruited from a local MVA site as adolescents successfully tested for provisional licenses. At the MVA, participating parents completed written surveys about expected teen driving during the 1st month of provisional licensure. One month later, 579 parent-teen dyads completed follow-up telephone interviews about teen driving within the past month. On weeks assigned as intervention, parents were exposed to a video and given the video and a driving agreement to take home. In multivariate linear regression analyses, the results indicated that when controlling for selected demographic and baseline psychosocial variables, intervention parents reported more driving rules, restricted driving, limits for high-speed roads, weekend night restrictions, and overall driving limits than did parents in the control group. When compared to control teens, intervention teens reported more limits on passengers, high-speed roads, and night driving, and on overall driving limits, but there were no differences for overall driving or driving under high-risk conditions. In addition, intervention parents were about 3 times, and intervention teens were about 5 times, more likely than controls to report using a parent-teen driving agreement. These results indicate that brief exposure to intervention at an MVA office may help increase parental limits on teen driving. PMID- 15134316 TI - Exploring the relationship between mental health and smoking cessation: a study of rural teens. AB - This study examined the association between mental health and smoking cessation among rural youth. Participants were 113 male and 145 female adolescents ages 14 19 from rural West Virginia and North Carolina. Participants were enrolled in the American Lung Association's 10-week Not On Tobacco (N-O-T) program or a 15-min single-dose brief intervention. Baseline and postprogram measures were completed on smoking status (i.e., quit, reduction), nicotine dependence, smoking history, and depression and anxiety. Results showed that more N-O-T participants quit and reduced smoking than did brief intervention participants. Intervention group, baseline smoking rate, and the Group x Gender, Group x Anxiety, and Group x Depression interactions were significant predictors of change in smoking behavior from baseline to postprogram. In conclusion, more N-O-T participants demonstrated favorable changes in smoking than did brief intervention participants. Approximately 1/3 of youth exhibited mental health pathology; more females than males. Levels of depression and anxiety improved from baseline to postprogram, overall. Although the extent of the impact of mental health on cessation outcomes was inconclusive, findings suggest that rural youth who smoke may be at risk for pathological depression and anxiety. Future cessation programming with rural youth should consider the inclusion of coping and stress management skills and mental health referral protocols as significant program components. PMID- 15134318 TI - Topical macrolactams: new indications, potential pitfalls. PMID- 15134317 TI - Tijuana alcohol control policies: a response to cross-border high-risk drinking by young Americans. AB - Several thousand young Americans visit the bars in Tijuana, Mexico, each weekend night, raising concerns on both sides of the border. Measures implemented in San Diego, California, and Tijuana have successfully reduced the number of American visitors to Mexican bars. Although San Diego policies have been well-documented, this is the first article on investigation of measures enacted south of the border. Information on Tijuana alcohol policies was obtained from a survey of 29 36 bars from 1997 to 1999. The Tijuana police provided data on Americans arrested in Tijuana from 1998 to 1999. Our study found alcohol regulations are poorly enforced in Tijuana, suggesting that regulatory agencies are captured by bar owners. However, such a capture may be weakening. The importance of identifying and supporting Mexican interest groups, as opposed to the bar owners, as a mechanism to impede the capture of Tijuana's regulatory agencies is discussed. The number of Americans involved in alcohol-related crimes in Tijuana sharply decreased over time. However, such a success is largely related to the success of the San Diego efforts in reducing the number of American visitors to Tijuana. Also, by demonstrating the racial/ethnic heterogeneity of American visitors to Tijuana bars, our study points out the need for prevention policies designed north of the border to take such heterogeneity into account. PMID- 15134319 TI - What is your diagnosis? Tinea in tattoo. PMID- 15134320 TI - Infantile myofibromatosis: a case report and review of the literature. AB - Infantile myofibromatosis is a rare mesenchymal disorder of infancy and childhood characterized by the formation of tumors in the soft tissues, muscle, bone, and viscera. Disease limited to the soft tissues, muscle, and bone has a good prognosis, and excision is curative; however, visceral involvement may be fatal. We present a case of infantile myofibromatosis in a 1-year-old boy and review the literature. PMID- 15134321 TI - What's eating you? Schistosoma haematobium. PMID- 15134322 TI - Tacrolimus-induced tinea incognito. AB - Tacrolimus and pimecrolimus represent a new class of topical nonsteroidal medications currently used in the treatment of a variety of inflammatory skin lesions. We report the case of a patient in whom topical tacrolimus therapy resulted in widespread lesions of tinea incognito. This case shows that partial treatment of dermatophytosis with griseofulvin may obscure the diagnosis. It also suggests that topical tacrolimus appears capable of inducing widespread dermatophytosis. The clinical appearance in this case was similar to tinea incognito induced by a topical corticosteroid. PMID- 15134323 TI - Diltiazem-induced hyperpigmentation. AB - A healthy-appearing 66-year-old black woman presented with a 5-year history of facial hyperpigmentation that was unresponsive to topical sunscreen, hydroquinone, tretinoin, and azelaic acid. Her medications included extended release diltiazem for hypertension for the past 7 years, rofecoxib for arthritis, and pantoprazole for esophagitis. On examination, the woman displayed hyperpigmented patches and papules involving most of her face. The punch biopsy findings from a hyperpigmented papule on the right temple revealed compact hyperkeratosis, follicular dilation, and dense inflammatory infiltrate along the dermal-epidermal junction with abundant dying keratinocytes. Her diltiazem therapy was discontinued, which led to gradual resolution of her hyperpigmentation. PMID- 15134324 TI - Disseminated cutaneous acanthamebiasis: a case report and review of the literature. AB - The genus Acanthamoeba includes species of free-living soil and water ameba that have been implicated in a small number of human diseases. Acanthamoeba species have been identified as the etiologic agents in 2 well-defined clinical entities, amebic keratitis and granulomatous amebic encephalitis (GAE). Less commonly, Acanthamoeba species have been identified as the cause of disseminated disease in debilitated and immunocompromised patients. Cutaneous acanthamebiasis, often a reflection of disseminated disease, is an increasingly recognized infection since the emergence of acquired immunodeficiency syndrome (AIDS) and the use of immunosuppressive drugs. The disease portends a poor prognosis and is uniformly fatal if the infection involves the central nervous system (CNS). We describe a patient with advanced AIDS who presented with disseminated cutaneous lesions, headache, and photophobia, and in whom a diagnosis of cutaneous acanthamebiasis was made based on the results of a skin biopsy. A multidrug therapeutic regimen was begun that included sulfadiazine; the patient responded favorably to treatment. This paper also reviews 36 previously reported cases of cutaneous acanthamebiasis with delineation of clinical, diagnostic, histologic, and prognostic features, as well as discusses treatment options. PMID- 15134325 TI - Cutis verticis gyrata secondary to a cerebriform intradermal nevus. AB - We report the case of a 30-year-old black man with a large mass consisting of longitudinal parallel ridges and furrows on the left parietal region. A small, single, hyperpigmented macule was present at birth and gradually grew and extended over the years. Cutis verticis gyrata was suspected, and an investigation was performed to discharge the possibility of pachydermoperiostosis. However, results of biopsies obtained from 3 different lesional areas showed the same histopathologic features--deep-seated hair follicles and clusters of nevus cells concentrated in the dermis. Cerebriform intradermal nevus is a rare cause of cutis verticis gyrata. Early diagnosis is extremely important to prevent the development of malignant melanoma. PMID- 15134326 TI - Keratoacanthoma centrifugum marginatum: a diagnostic and therapeutic challenge. AB - A keratoacanthoma centrifugum marginatum (KACM) may pose a diagnostic and therapeutic challenge. Clinically and histologically, it may resemble mycobacterial or deep fungal infection or halogenoderma. Therapy can be challenging because the lesion can expand to a great size. We report on a patient with multiple lesions of KACM. The diagnostic difficulty and the therapeutic failure of imiquimod, intralesional methotrexate (MTX), and isotretinoin, as well as the therapeutic success of 5-fluorouracil (5-FU) cream, are discussed. PMID- 15134327 TI - Tacrolimus ointment in the treatment of eyelid dermatitis. AB - The safety and efficacy of tacrolimus ointment 0.1% (Protopic) in the treatment of atopic dermatitis of the eyelids were assessed in an open-label clinical trial of 21 patients with moderate to severe eyelid dermatitis. Of those 21 patients, 20 received study drug and were followed. Patients applied tacrolimus ointment 0.1% twice daily for 8 weeks and were followed for 2 additional weeks after the last day of treatment. Complete eye examinations were conducted throughout the study. Efficacy was assessed through the investigator's evaluation of the patients' individual signs and symptoms of eyelid dermatitis and the physician global assessment (PGA) of eyelid clinical response. Improvement in the investigator's evaluation of the signs and symptoms of eyelid dermatitis was observed during the study. A total of 80% of patients (16/20) experienced marked improvement or better in PGA at 8 weeks. Adverse events were limited to local burning and itching after the first few applications of study medication. Of the 20 patients, 12 reported burning (60%), and 5 reported itching (25%). There was no statistically significant increase in intraocular pressure (IOP) during the study compared with baseline. In addition, none of the patients developed cataracts or glaucoma during the study. In summary, tacrolimus ointment 0.1% may be a safe and effective treatment option for patients with moderate to severe eyelid dermatitis. PMID- 15134328 TI - A morphological study on the human obturator externus muscle with reference to anomalous muscle and anomalous fasciculus originating from the obturator externus muscle. AB - To elucidate the essential nature of the human obturator externus muscle (OE), the gross anatomy of the anomalous muscle and anomalous fasciculus originating from this muscle as well as the posterior division of the obturator nerve (P) were examined in 73 thighs of 45 Japanese adult cadavers. The investigation of anomalous muscle and anomalous fasciculus was effected by observation of the supernumerary muscular branch and supernumerary muscular twig from the P. The anomalous muscle occurring between the adductor brevis muscle (AB) and adductor minimus muscle (AMi) (37/73 thighs; 50.7%) and the anomalous fasciculus fused to the posterior surface of the AMi (18/73 thighs; 24.7%) were detected. It was demonstrated by the course of the P and the suppling nerve, that the OE was divided into its superior fasciculus (or fasciculi) (SF) and main belly by the P, moreover the anomalous muscle originated from the former and the anomalous fasciculus from the latter. Presumably, in the process of ontogeny, the SF of the original OE retained its original morphology (23/73 thighs; 31.5%), a part of the SF was converted to anomalous muscle (27/73 thighs; 37.0%), the entire SF was converted to anomalous muscle (10/73 thighs; 13.7%), and the entire SF underwent retrogression and disappeared (13/73 thighs; 17.8%). The presence (50/73 thighs; 68.5%) or absence (23/73 thighs; 31.5%) of piercing of the OE by the P seemed to depend upon the circumstances surrounding the morphological change in the SF. The original OE was considered to occur as an OE pierced by the P not possessing any supernumerary muscular branch or twig. Actually, the original OE was confirmed to occur with a low frequency (15/73 thighs; 20.5%). It was inferred that the original OE readily underwent variation in the process of ontogeny due to the influence of the obturator nerve. The segmental composition of the obturator nerve was considered not to be involved in the morphological change in the OE. PMID- 15134329 TI - Fiber dissection technique for demonstrating the lateral lemniscus of the human brain. AB - This report describes fiber dissection technique for tracing the auditory pathway from the cochlear nerve to the medial geniculate body via the lateral lemniscus, inferior colliculus and inferior brachium. Some fibers of the lateral leminiscus appear to reach the thalamus in conjunction with fibers of the medial lemniscus. PMID- 15134330 TI - Lateral expansion of the rat lower dental arch by applying an active plate model to the mandibular first molar region: observation of histological changes in the lower alveolar bone over a 2-week period. AB - Lateral expansion of the lower dental arch has rarely been conducted clinically because the structure of the lower jaw is considered to be unsuitable for this type of treatment. However, successful lateral expansion of the lower dental arch using SCHWALZ, an orthopedic appliance has been reported in recent years. Therefore, an experimental study was performed to examine the histological changes in the lower alveolar bone when lateral expansion is applied to the lower dental arch for periods of up to 2 weeks. An active plate model based on the Schwarz appliance was attached to the first molar region in rats. The distance between the right and left first molars was measured, and the mandible and first molar tooth were processed for light microscopy at different times after the fitting. The lateral expansion caused by lateral movement of the right and left first molars following the application of the active plate amounted to an average of 1.06 mm over a period of 14 days. Alkaline phosphatase/tartrate-resistant acid phosphatase staining revealed bone deposition on the periosteal surface of the buccal alveolar bone and the periodontal surface of the lingual alveolar bone, whereas bone absorption was observed on the periodontal surface of the buccal alveolar bone and the periosteal surface of the lingual alveolar bone. These findings demonstrate that lateral expansion of the lower dental arch through the application of an active plate model was achieved by bony deposition and absorption of alveolar bone, similar to the process that occurs in association with lateral expansion of the upper dental arch. PMID- 15134331 TI - Effects of mono(2-ethylhexyl) phthalate (MEHP) on testes in rats in vitro. AB - The phthalate esters have been used as plasticizers for various plastic products, and their testicular toxicity has been reported. In this study, the effects of mono(2-ethylhexyl) phthalate (MEHP), one of the phthalate esters, on prepubertal rat testes in vitro were examined. The testes of 20-day-old Sprague Dawley (SD) rats were cut into smaller pieces and seeded in medium, and then the specimens were obtained for light and transmission electron microscopic observations. As a result, at 1 hr after exposure to MEHP, TUNEL-positive spermatogenic cells were identified, and they gradually increased in number in time- and dose-dependent manners. Ultrastructurally, apoptotic spermatogenic cells (characterized with chromatin condensation, cytoplasm shrinkage without membrane rupture, still functioning cell organelles, and packed cell contents in membrane-bounded bodies), necrotic spermatogenic cells (characterized with swollen and ruptured mitochondria, plasma membrane lysis, spilt cell contents, and chromatin clumps), apoptotic Sertoli cells (highly condensed nuclei and nuclear membrane lysis) and necrotic Sertoli cells (marginated chromatins along the nuclear membrane, some swollen and ruptured cell organelles, e.g. mitochondria) could be identified. Conclusively, based on transmission electron microscopic observations, MEHP treatment may affect spermatogenic cells, and lead them to necrosis. Thus, testicular tissue cultures and cell cultures are of advantageous for screening testicular toxicity of chemicals. PMID- 15134332 TI - Oxygen and the cell. PMID- 15134333 TI - O2 sensing in the human ductus arteriosus: redox-sensitive K+ channels are regulated by mitochondria-derived hydrogen peroxide. AB - The ductus arteriosus (DA) is a fetal artery that allows blood ejected from the right ventricle to bypass the pulmonary circulation in utero. At birth, functional closure of the DA is initiated by an O2-induced, vasoconstrictor mechanism which, though modulated by endothelial-derived endothelin and prostaglandins, is intrinsic to the smooth muscle cell (DASMC) [Michelakis et al., Circ. Res. 91 (2002); pp. 478-486]. As pO2 increases, a mitochondrial O2 sensor (electron transport chain complexes I or III) is activated, which generates a diffusible redox mediator (H2O2). H2O2 inhibits voltage-gated K+ channels (Kv) in DASMC. The resulting membrane depolarization activates L-type Ca2+ channels, thereby promoting vasoconstriction. Conversely, inhibiting mitochondrial ETC complexes I or III mimics hypoxia, depolarizing mitochondria, and decreasing H2O2 levels. The resulting increase in K+ current hyperpolarizes the DASMC and relaxes the DA. We have developed two models for study of the DA's O2-sensor pathway, both characterized by decreased O2-constriction and Kv expression: (i) preterm rabbit DA, (ii) ionically-remodeled, human term DA. The O2-sensitive channels Kv1.5 and Kv2.1 are important to DA O2-constriction and overexpression of either channel enhances DA constriction in these models. Understanding this O2-sensing pathway offers therapeutic targets to modulate the tone and patency of human DA in vivo, thereby addressing a common form of congenital heart disease in preterm infants. PMID- 15134334 TI - Oxidative stress in the systemic and cellular responses to intermittent hypoxia. AB - Patients with chronic intermittent hypoxia (IH) caused by recurrent apneas have a greatly increased risk for developing hypertension, myocardial infarctions, and stroke. The purpose of this article is to highlight some of the recent studies focusing on the mechanisms associated with systemic and cellular effects of IH in experimental animals and cell culture models. Rats exposed to chronic IH exhibited elevated blood pressures and increased sympathetic nerve activity, partly due to enhanced reflexes arising from carotid bodies. Direct recordings of the carotid body sensory activity showed that chronic IH selectively augmented hypoxic sensitivity, and induced a novel form of functional plasticity manifested as sensory long-term facilitation. In cell culture models, prior exposure to IH resulted in facilitation of hypoxia-induced transmitter release and activation of several protein kinases. IH caused activation of c-Fos and activator protein-1 (AP-1) transcription factor and tyrosine hydroxylase, an AP-regulated downstream gene. For a given duration and intensity of hypoxia, IH was more potent and caused longer-lasting activation than continuous hypoxia. Scavengers of reactive oxygen species (ROS) prevented IH-induced systemic and cellular responses. Inhibition of complex I of the mitochondrial electron transport chain appears to be one of the sources for IH-induced generation of ROS. The persistent oxidative stress may contribute to the progression of morbidity associated with chronic IH caused by recurrent apneas, and antioxidants might be of considerable therapeutic value in preventing the progression of disease associated with chronic IH. PMID- 15134335 TI - HIF hydroxylation and cellular oxygen sensing. AB - Hypoxia-inducible factor (HIF) is a transcriptional complex that mediates a broad range of cellular and systemic responses to hypoxia. Analysis of HIF-alpha subunits has demonstrated that its activity is regulated by a series of oxygen dependent enzymatic hydroxylations at specific prolyl and asparaginyl residues. Combined structural/genetic approaches have identified the relevant enzymes as members of the 2-oxoglutarate-dependent dioxygenase superfamily, possessing a beta-barrel 'jelly-roll' conformation that aligns a 2-histidine/1-carboxylate iron co-ordination motif at the catalytic centre. HIF prolyl hydroxylation is performed by a closely related set of isoenzymes (PHD1-3) that differ in abundance and subcellular localisation. Hydroxylation of either human HIF-1alpha Pro402 or Pro564 promotes interaction with the von Hippel-Lindau tumour suppressor protein (pVHL). In oxygenated cells this process targets HIF-alpha for rapid proteasomal destruction. HIF asparaginyl hydroxylation is performed by a protein termed factor inhibiting HIF (FIH). In oxygenated cells hydroxylation of human HIF-1alpha Asn803 prevents interaction with the p300 transcriptional co activator, providing a second mechanism by which HIF-mediated transcription is inactivated. Genetic studies demonstrate a critical function for both types of enzyme in regulating the HIF transcriptional cascade. Limitation of activity in hypoxia supports a central role of these hydroxylases in cellular oxygen sensing. Regulation of the amount of hydroxylase protein, and the supply of other co substrates and co-factors, particularly the cellular availability of iron, also contribute to tuning the physiological response to hypoxia. PMID- 15134336 TI - Visualization of the three-dimensional organization of hypoxia-inducible factor-1 alpha and interacting cofactors in subnuclear structures. AB - Cells need oxygen (O2) to meet their metabolic demands. Highly efficient systems of O2-sensing have evolved to initiate responses enabling cells to adapt their metabolism to reduced O2 availability. Of central importance is the activation of hypoxia-inducible factor-1 (HIF-1), a transcription factor complex that controls the expression of genes the products of which regulate glucose uptake and metabolism, vasotonus and angiogenesis, oxygen capacity of the blood as well as cell growth and death. Activation of HIF-1 requires the accumulation and nuclear translocation of the HIF-1alpha subunit, its dimerization with HIF-1beta and the binding of co-activator proteins such as p300. In this study we investigated the three-dimensional (3D) distribution of HIF-1alpha within the nucleus and assigned its localization to known nuclear compartments. Using two-photon microscopy we determined the colocalization of HIF-1alpha and -beta subunits within nuclear domains as well as overlaps between HIF-1alpha and p300. Our data provide information on the nuclear distribution of HIF-1alpha with respect to subnuclear domains that could serve as specific locations for hypoxia-induced gene expression. PMID- 15134337 TI - Modulation of glucokinase expression by hypoxia-inducible factor 1 and upstream stimulatory factor 2 in primary rat hepatocytes. AB - Glucokinase (GK) is the key enzyme of glucose utilization in liver and is localized in the less aerobic perivenous area. Until now, the O2-responsive elements in the liver-specific GK promoter are unknown, and therefore the aim of this study was to identify the O2-responsive element in this promoter. We found that the GK promoter sequence -87/-80 matched the binding site for hypoxia inducible factor 1 (HIF-1) and upstream stimulatory factor (USF). In primary rat hepatocytes we could show that venous pO2 enhanced HIF-1alpha and USF-2a levels, both of which activated GK expression. Furthermore, transfection experiments revealed that the GK sequence -87/-80 mediated the HIF-1alpha- or USF-2-dependent activation of the GK promoter. The binding of HIF-1 and USF to the GK-HRE was corroborated by electrophoretic mobility shift assay (EMSA). However, the maximal response to HIF-1alpha or USF was only achieved when constructs with the -87/-80 sequence in context with a 3'-36 bp native GK promoter sequence containing a hepatocyte nuclear factor 4 (HNF-4) binding site were used. HIF-1alpha and HNF-4 additively activated the GK promoter, while USF-2 and HNF-4 together did not show this additive activation. Thus, HIF-1 and USF may play differential roles in the modulation of GK expression in response to O2. PMID- 15134338 TI - Redox-sensitive regulation of the HIF pathway under non-hypoxic conditions in pulmonary artery smooth muscle cells. AB - Pulmonary hypertension and vascular remodeling processes are associated with oxidative stress, hypoxia and enhanced levels of thrombin and vascular endothelial growth factor (VEGF). The hypoxia-inducible transcription factor HIF regulates the expression of VEGF under hypoxia. The HIF pathway is also activated by thrombin or CoCl2, likely via reactive oxygen species (ROS). In this study we investigated whether the redox-modifying enzymes superoxide dismutase (SOD), glutathione peroxidase (GPX) and catalase affect HIF levels and the expression of VEGF mRNA in pulmonary artery smooth muscle cells (PASMC). Stimulation of PASMC with thrombin or CoCl2 increased ROS production and enhanced HIF-alpha protein and VEGF mRNA levels as well as HIF-dependent reporter gene activity. These responses were inhibited by vitamin C and by overexpression of GPX and catalase, whereas the opposite effects were observed in SOD-expressing cells. These findings suggest that an 'antioxidant' state with reduced levels of H2O2 limits the activation of the HIF pathway, whereas a 'prooxidant' state allowing elevated H2O2 levels promotes it. Thus, shifting the redox balance to a more reduced environment, thereby limiting VEGF expression, may be beneficial for treating remodeling processes during pulmonary hypertension. PMID- 15134339 TI - Measurement of exhaled hydrogen peroxide from rabbit lungs. AB - Exhaled H2O2 is considered an indicator of lung inflammatory and oxidative stress. Moreover, H2O2 may be involved in signal transduction processes. It is not fully elucidated to what extent (i) H2O2 escapes from the intravascular compartment, and (ii) pulmonary H2O2 generation and nasopharyngeal H2O2 generation contribute to exhaled H2O2. We investigated H2O2 concentrations in breath condensate from isolated buffer-perfused and ventilated rabbit lungs, and from both intubated and spontaneously breathing rabbits with a horseradish peroxidase/2',7'dichlorofluorescin assay. For the perfused lungs, a H2O2 concentration of 58 +/- 19 nM was found. Addition of H2O2 to the buffer fluid resulted in only minute appearance in the exhaled air (<0.001%). Levels of exhaled H2O2 in intubated rabbits and perfused lungs were virtually identical. Nearly ten-fold higher levels were detected in spontaneously breathing rabbits. Decreasing the inspired oxygen concentration from 21% to 1% resulted in a tendency toward decreased H2O2 exhalation in perfused lungs. In contrast, phorbol 12-myristate-13-acetate (PMA) prompted a approximately 4-fold increase in H2O2 exhalation. We conclude that the horseradish peroxidase/2',7'dichlorofluorescin assay is a feasible technique to measure H2O2 in exhaled breath condensate in rabbits. When collecting exhaled air via the tracheal tube, the signal represents pulmonary H2O2 generation with the contribution of the remaining body being negligible. PMID- 15134340 TI - Effects of reducing agents on glutathione metabolism and the function of carotid body chemoreceptor cells. AB - Two current hypotheses of O2 sensing in the carotid body (CB) chemoreceptors suggest participation of oxygen reactive (ROS) species, but they are mechanistically opposed. One postulates that hypoxia decreases ROS levels; the other that hypoxia increases them. Yet, both propose that the ensuing alteration in the cellular redox environment is the key signal triggering hypoxic chemoreception. Since the glutathione redox pair is the main cellular buffer for ROS and the main determinant of the general redox environment of the cells, a way to test whether ROS participate in chemoreception is to determine glutathione levels and to correlate them with the activity of CB chemoreceptor cells. We found that hypoxia does not alter the glutathione reduction potential but that it activates chemoreceptor cell neurosecretion. Incubation of tissues with reduced glutathione increases the glutathione-reducing potential but does not activate chemoreceptor cells in normoxia nor does it modify hypoxic activation. Like reduced glutathione, N-acetylcysteine promoted a general reducing environment in the cells without alteration of chemoreceptor cell activity. N (mercaptopropionyl)-glycine, like the two previous agents, increases the reduction potential of glutathione. In contrast, the compound activated chemoreceptor cells in normoxia, promoting a dose- and Ca(2+)-dependent neurosecretion and a potentiation of the hypoxic responses. The existence of multiple relationships between glutathione reduction potential in the cells and their activity indicates that the general cellular redox environment is not a factor determining chemoreceptor cell activation. It cannot be excluded that the local redox environments of restricted microdomain(s) in the cells with specific regulating mechanisms are important signals for chemoreceptor cell activity. PMID- 15134341 TI - Expression of functional purinergic receptors in pulmonary neuroepithelial bodies and their role in hypoxia chemotransmission. AB - Adenine nucleotides act through specific cell surface receptors to invoke a variety of biological responses. Here we show that cells of neuroepithelial bodies (NEB), presumed O2 airway sensors in neonatal hamster lung, express functional P2X receptors (P2X-R). Positive immunostaining was detected in NEB cells using double-label immunohistochemistry with antibodies against P2X2 and P2X3 receptor subunits, which co-localized with serotonin (5-HT), a marker of NEB cells. For electrophysiological characterization of P2X2-R in NEB cells, fresh neonatal hamster lung slice preparation was used. Under whole-cell patch clamp, perfusion with ATP induced a concentration-dependent, non-desensitizing inward current (EC50=12 microM). Perfusion with alpha,beta-methylene ATP also induced a slow-desensitizing inward current (EC50=8.2 microM). Suramin (IC50 ca. 43 microM) and TNP-ATP (IC50 ca. 8 microM) blocked the currents evoked by both ATP and alpha,beta-methylene ATP. Using carbon fiber amperometry we observed that hypoxia and ATP induced 5-HT release from NEB cells and that this release was blocked by suramin. These data suggest that functional P2X2/3 heteromeric receptors are expressed in NEB cells. The possible function of these purinoreceptors in NEB cells could include modulation of hypoxia chemotransmission. PMID- 15134343 TI - Simultaneous exposure of rats to dioxin and carbon monoxide reduces the xenobiotic but not the hypoxic response. AB - Aryl hydrocarbon receptor (AhR) and hypoxia-inducible factor-1alpha (HIF-1alpha) are conditionally regulated transcription factor subunits that form heterodimeric complexes with their common partner, AhR nuclear translocator (ARNT/HIF-1beta). Whereas the environmentally toxic compound 2,3,7,8-tetra-chlorodibenzo-p-dioxin (TCDD) initiates the trans-activation activity of AhR:ARNT/HIF-1beta, hypoxic exposure stabilizes HIF-1alpha and functionally activates the HIF-1alpha:ARNT/HIF 1beta complex. To analyze a possible crosstalk between these two pathways in vivo, rats were given dioxin orally and/or were exposed to carbon monoxide (CO), causing functional anemia. We found that exposure to CO inhibited the xenobiotic response while dioxin application had no significant negative impact on hypoxia mediated gene transcription. PMID- 15134342 TI - Remodelling of Ca2+ homeostasis in type I cortical astrocytes by hypoxia: evidence for association with Alzheimer's disease. AB - Sustained central hypoxia predisposes individuals to dementias such as Alzheimer's disease, in which cells are destroyed in part by disruption of Ca2+ homeostasis. Here, we show that exposure of astrocytes to hypoxia in vitro causes inhibition of plasmalemmal Na+/Ca2+ exchange and excessive mitochondrial Ca2+ loading. Both factors disrupt normal agonist-evoked Ca2+ signalling. Moreover, hypoxia increases the levels of presenilin-1, a major component of a key enzyme involved in Alzheimer's disease. Inhibition of this enzyme partially reverses the effects of hypoxia on Ca2+ signalling. These findings provide an initial cellular basis for understanding the clinical association of hypoxia with Alzheimer's disease. PMID- 15134344 TI - Structure and expression of two kininogen genes in mice. AB - Kininogens serve dual functions by forming a scaffold for the assembly of the protein complex initiating the surface-activated blood coagulation cascade and as precursors for the kinin hormones. While rats have three kininogen genes, for mice, cattle, and humans only one gene has been described. Here, we present sequence and expression data of a second mouse kininogen gene. The two genes, kininogen-I and kininogen-II, are located in close proximity on chromosome 16 in a head-to-head arrangement. In liver and kidney, both genes are expressed and for each gene three alternative splice variants are synthesized. Two of them are the expected high and low molecular weight isoforms known from all mammalian kininogens. However, for both genes also a third, hitherto unknown splice variant was detected which lacks part of the high molecular weight mRNA due to splicing from the low molecular weight donor site to alternative splice acceptor sites in exon 10. The physiological functions of the six kininogen isoforms predicted by these findings need to be determined. PMID- 15134345 TI - The central domain of the matrix protein of HIV-1: influence on protein structure and virus infectivity. AB - The central region of the matrix protein p17 of HIV-1 is known to be essential during virus assembly. We substituted alanines for amino acid triplets in this region of p17 (amino acid residues 47 to 55: NPG LLE TSE). Introduction of the respective mutations into the gag-coding sequence of HI-proviruses and subsequent transfection into Cos-7 cells led to particle production and release. Exchange of LLE resulted in the production of non-infectious particles. These residues may be important for correct folding and assembly of the processed matrix protein and the production of infectious HIV. In vitro studies of wild-type and mutated matrix proteins using spectroscopic methods (NMR, fluorescence, CD) yielded detailed data about structure and stability. Two-dimensional NMR spectroscopy showed that wild-type and mutant proteins (p17-NPG and p17-TSE) are well folded. Besides structural changes at the mutated site, chemical shift changes indicate small but significant long range structural rearrangements. The stability against chemically and thermally induced unfolding of the mutants p17-NPG and p17-TSE was slightly decreased, while that of p17-LLE was drastically diminished. The alterations have only a local effect on protein folding for the mutants p17-NPG and p17-TSE, and the globular tertiary structure remains nearly unchanged. For p17-LLE, however, the substitutions seem to trigger significant changes in structural elements. PMID- 15134346 TI - Skin secretion of the toad Bombina variegata contains multiple insulin-releasing peptides including bombesin and entirely novel insulinotropic structures. AB - Skin secretions of the toad Bombina variegata were evaluated for the isolation and characterisation of insulinotropic peptides. Crude secretions obtained from young adult toads by mild electrical stimulation of the dorsal skin surface were purified by reverse phase HPLC yielding 44 peaks. In acute incubations with glucose-responsive BRIN-BD11 cells, peaks 21, 22, 23, 24 and 25 showed a 1.5-3.5 fold increase in insulin release compared with 5.6 mM glucose alone (p<0.001; n=3). Structural analyses of the purified insulin-releasing peaks were performed by automated Edman degradation and mass spectrometry. Peptides represented by peaks 21, 22 and 23 had molecular masses of 1641.7 Da, 1662.6 Da and 1619.8 Da respectively. These peptides were unblocked by removal of pyroglutamic acid from the N-terminus prior to Edman degradation, revealing lengths of 14 amino acids. Peak 21 yielded a primary structure of Pyr-QRLGHQWAVGHLM, which a data base search revealed as an analogue of bombesin (His6 bombesin), while peak 23 was an exact match of bombesin (Pyr-QRLGNQWAVGHLM) originally isolated from Bombina bombina. Peak 22 indicated a primary structure of Pyr-DSFGNQWARGHFM (72% homology with bombesin). Peaks 24 and 25 revealed entirely novel insulinotropic peptides with molecular masses and primary structures of 1650.5 Da and 2300.0 Da and GKPFYPPPIYPEDM (GM-14) and IYNAICPCKHCNKCKPGLLAN (IN-21) respectively. Preliminary studies on the mechanisms underlying the insulinotropic actions of peaks 21, 22, 23 and 24 suggest possible involvement of a cAMP-dependent, G protein-insensitive pathway. These data indicate that Bombina variegata skin secretions contain peptides with insulin-releasing activity, which may have mammalian counterparts and prove useful for possible exploitation as antidiabetic agents from natural resources. PMID- 15134347 TI - Inhibition of lentil copper/TPQ amine oxidase by the mechanism-based inhibitor derived from tyramine. AB - Copper amine oxidase from lentil (Lens esculenta) seedlings was shown to catalyze the oxidative deamination of tyramine and three similar aromatic monoamines, benzylamine, phenylethylamine and 4-methoxyphenylethylamine. Tyramine, an important plant intermediate, was found to be both a substrate and an irreversible inhibitor of the enzyme whereas the other amines were not inhibitory. In the course of tyramine oxidation the enzyme gradually became inactivated with the concomitant appearance of a new absorption at 560 nm due to the formation of a stable adduct. Inactivation took place only in the presence of oxygen and was probably due to the reaction of the enzyme with the oxidation product of tyramine, p-hydroxyphenylacetaldehyde. The kinetic data obtained in this study indicate that tyramine represents a new interesting type of physiological mechanism-based inhibitor for plant copper amine oxidases. PMID- 15134348 TI - Enhanced expression of basolateral multidrug resistance protein isoforms Mrp3 and Mrp5 in rat liver by LPS. AB - Lipopolysaccharide (LPS) induces hepatocellular down-regulation and endocytic retrieval of multidrug resistance protein 2 (Mrp2, Abcc2). Basolateral Mrp isoforms may compensate for the intracellular metabolic changes in cholestasis. Therefore, the effect of LPS on the zonal localization of Mrp2 and Mrp3 and the expression of Mrp3, Mrp4, Mrp5, and Mrp6 mRNA were investigated in rat liver. In normal rat liver Mrp3 was found in pericentral hepatocytes also expressing glutamine synthetase. In LPS-treated rat liver the decrease in Mrp2 protein was most pronounced in pericentral hepatocytes, with only minor down-regulation in periportal hepatocytes. Conversely, induction of Mrp3 was found in pericentral hepatocytes with a low expression of Mrp2. Furthermore, we found a strong induction of Mrp5 mRNA. Likewise, Mrp6 mRNA was up-regulated, however Mrp6 protein expression was not significantly altered. It is concluded that Mrp3 is inversely regulated to Mrp2 in a zonal pattern and may compensate for the LPS induced loss of Mrp2 in the perivenous area. Induction of pericentral Mrp3 and up regulation of Mrp5 mRNA may play an important role in the hepatocellular clearance of cholephilic substances and cyclic nucleotides accumulating after LPS treatment. PMID- 15134349 TI - Critical O2 and NO concentrations in NO-induced cell death in a rat liver sinusoidal endothelial cell line. AB - Nitric oxide (NO) plus oxygen (O2) are known to cause cell damage via formation of reactive nitrogen species. NO itself directly inhibits cytochrome oxidase of the mitochondrial respiratory chain in competition with O2, thus inducing a hypoxic-like injury. To assess the critical NO and O2 concentrations for both mechanisms of NO-induced cell injury, cells of a rat liver sinusoidal endothelial cell line were incubated in the presence of the NO donor spermineNONOate at different O2 concentrations, and their loss of viability was determined by the release of lactate dehydrogenase. Protection by ascorbic acid was used as indication for the involvement of reactive nitrogen species, whereas a hypoxic like injury was indicated by the protective effects of glycine and glucose and the increase in NAD(P)H fluorescence. High concentrations of NO (approx. 10 microM NO) and O2 (21% O2) were required to induce endothelial cell death mediated by formation of reactive nitrogen species. On the other hand, pathophysiologically relevant NO concentrations at low but physiological O2 concentrations (ca. 2 microM NO at 5% O2 and about 1 microM NO at 2% O2) induced hypoxic-like cell death in the endothelial cells that was prevented by the presence of glucose. PMID- 15134350 TI - A virtual cornucopia of alternative therapies. PMID- 15134351 TI - Neoadjuvant endocrine therapy for breast cancer: an overlooked option? AB - For many oncologists, neoadjuvant treatment for breast cancer is synonymous with preoperative cytotoxic chemotherapy, regardless of tumor characteristics. Preoperative therapy with an endocrine agent is generally considered suitable only for the frail elderly or the medically unfit. However, favorable information regarding third-generation aromatase inhibitors in the treatment of all stages of breast cancer prompts a reconsideration of this bias. In light of the fact that neoadjuvant therapy with aromatase inhibitors is restricted to postmenopausal women with strongly estrogen-receptor-positive tumors, the assumption that neoadjuvant combination chemotherapy is more efficacious than a third-generation aromatase inhibitor can be reasonably questioned. It is particularly remarkable that the outcome of a comparison of adjuvant tamoxifen vs anastrozole (Arimidex)- the Arimidex, Tamoxifen Alone or in Combination (ATAC) trial--in more than 6,000 patients was predicted by a neoadjuvant trial that showed an efficacy advantage for a third-generation aromatase inhibitor (letrozole [Femara]) compared to tamoxifen in a sample of 337 patients after only 4 months of treatment. The potential of the neoadjuvant setting in efforts to identify new biologic agents that could build on the effectiveness of adjuvant aromatase inhibitors is therefore beginning to be appreciated. Finally, neoadjuvant therapy with an aromatase inhibitor could be considered a sensitivity test of endocrine therapy that might be incorporated into strategies to individualize treatment according to response. For this possibility to be realized, however, a better understanding of the relationship between surrogates from the neoadjuvant setting and the long term outcome of adjuvant aromatase inhibitor therapy will have to be established through practice-setting clinical trials. PMID- 15134352 TI - Case 3: infiltrating ductal breast carcinoma. PMID- 15134353 TI - Patient selection for prostate brachytherapy: more myth than fact. AB - Prostate brachytherapy efficacy and morbidity are primarily dependent on implant quality and patient management. While most alleged contraindications to brachytherapy have been propagated based on physician bias rather than clinical data, a number of evidence-based factors contributing to radiation-related morbidity have accumulated. Refinements in patient selection may result in further improvements in biochemical and quality-of-life outcomes. Herein, we have summarized evidence-based vs unsubstantiated patient selection factors that affect outcome. PMID- 15134354 TI - Clinical trials referral resource. Current phase III, NCI-supported, Cooperative Group Clinical Trials in breast cancer. PMID- 15134355 TI - Ovarian immature teratoma with implants and metastases. PMID- 15134356 TI - Epidermal growth factor receptor inhibitors for the treatment of colorectal cancer: a promise fulfilled? AB - The epidermal growth factor receptor (EGFR) is commonly expressed in colorectal cancers but not in most normal tissues, raising the possibility that this receptor could serve as a target for highly selective therapy. Based on preclinical studies demonstrating that antagonists of EGFR resulted in the inhibition of tumor growth, the development of clinical reagents has been aggressively pursued. Early clinical studies demonstrated antitumor activity of EGFR inhibitors in patients with advanced colorectal cancer, with acceptable toxicity. This early success fueled rapid clinical development. In this article, we will review the current status of EGFR inhibitors in the treatment of patients with colorectal cancer, in an effort to describe both how far we have come as well as where we need to go in optimizing this promising therapeutic approach. PMID- 15134357 TI - Proteomics to diagnose human tumors and provide prognostic information. AB - Proteomics is a rapidly emerging scientific discipline that holds great promise in identifying novel diagnostic and prognostic biomarkers for human cancer. Technologic improvements have made it possible to profile and compare the protein composition within defined populations of cells. Laser capture microdissection is a tool for procuring pure populations of cells from human tissue sections to be used for downstream proteomic analysis. Two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) has been used traditionally to separate complex mixtures of proteins. Improvements in this technology have greatly enhanced resolution and sensitivity providing a more reproducible and comprehensive survey. Image analysis software and robotic instrumentation have been developed to facilitate comparisons of complex protein expression patterns and isolation of differentially expressed proteins spots. Differential in-gel electrophoresis (DIGE) facilitates protein expression by labeling different populations of proteins with fluorescent dyes. Isotope-coded affinity tagging (ICAT) uses mass spectroscopy for protein separation and different isotope tags for distinguishing populations of proteins. Although in the past proteomics has been primarily used for discovery, significant efforts are being made to develop proteomic technologies into clinical tools. Reverse-phase protein arrays offer a robust new method of quantitatively assessing expression levels and the activation status of a panel of proteins. Surface-enhanced laser-desorption/ionization time-of-flight (SELDI-TOF) mass spectroscopy rapidly assesses complex protein mixtures in tissue or serum. Combined with artificial intelligence-based pattern recognition algorithms, this emerging technology can generate highly accurate diagnostic information. It is likely that mass spectroscopy-based serum proteomics will evolve into useful clinical tools for the detection and treatment of human cancers. PMID- 15134358 TI - Informatically challenged. PMID- 15134359 TI - Natural and experimental Helicobacter infections. AB - Current information about Helicobacter infections in humans and various domestic, wild, and research animal species that have been used or have the potential to be used as animal models of human disease is presented. The Helicobacter genus now includes at least 26 formally named species, with additional novel species in the process of being characterized. The natural history, host range with zoonosis potential, pathology, and diagnostic techniques are presented, along with examples of how Helicobacter infection has interfered with unrelated in vivo research. Current recommendations for deriving and managing helicobacter-free animal colonies for research are provided. PMID- 15134360 TI - Minimally invasive surgery via laparoscopy for intra-abdominal biopsy in obese rhesus macaques (Macaca mulatta). AB - PURPOSE: To determine the safety and effectiveness of laparoscopy for repeated intra-abdominal biopsy of liver and omental adipose tissue (AT) in obese rhesus monkeys (Macaca mulatta). METHODS: Nine obese rhesus monkeys were studied by use of 18 laparoscopic procedures (two procedures each, approx. six weeks apart). Time-sensitive liver and omental AT specimens were obtained from monkeys under general anesthesia, using a three-port approach with a roticulating endoscopic stapler/divider and a monopolar electrosurgery for hemostasis. RESULTS: All subjects tolerated the initial and repeat laparoscopic procedures well. Liver specimens weighed a mean +/- SEM of 3.8 +/- 0.5 g, and omental AT specimens weighed 16.6 +/- 0.8 g. Compared with previous studies of conventional laparotomy with liver wedge resection, the monkeys experienced faster postoperative recovery via laparoscopy, with rapid return to normal food intake and activity. Minimal to no adhesions were observed by use of the repeat procedure in all monkeys, with no major complications. CONCLUSIONS: Laparoscopy in obese rhesus monkey (ranging from young to older-aged), with repeated intra-abdominal liver and omental AT biopsy, was an excellent minimally invasive surgical method. In contrast to laparotomy with wedge resection, this approach greatly decreases operative time and stress, provides generous tissue specimens in a time-efficient manner, and facilitates rapid and full recovery of the nonhuman primate. PMID- 15134361 TI - Metabolism of daidzein by intestinal bacteria from rhesus monkeys (Macaca mulatta). AB - PURPOSE: To identify the metabolites produced from an isoflavonoid, daidzein, by colonic bacteria of rhesus monkeys. METHODS: The metabolism of daidzein by the fecal bacteria of nine monkeys was investigated. Daidzein was incubated anaerobically with fecal bacteria, and the metabolites were analyzed by use of liquid chromatography and mass spectrometry. RESULTS: The fecal bacteria of all of the monkeys metabolized daidzein to various extents. Dihydrodaidzein was found in cultures of fecal bacteria from two monkeys; dihydrodaidzein and equol were found in cultures from four monkeys; dihydrodaidzein, equol, and an unknown metabolite (MW = 244) were found in cultures from one monkey; and dihydrodaidzein and the unknown metabolite were found in cultures from two monkeys. CONCLUSIONS: Similar to that in humans, variation was evident in the metabolism of isoflavonoids by fecal bacteria from rhesus monkeys. Some metabolites produced by fecal bacteria from monkeys were the same as those produced by fecal bacteria from humans. PMID- 15134362 TI - Effect of water hardness on oocyte quality and embryo development in the African clawed frog (Xenopus laevis). AB - Husbandry and health of the African clawed frog, Xenopus laevis, greatly influences the quality of oocytes produced. One factor affecting oocyte quality is the water conditions in which females are maintained. Dechlorination and adequate salt concentration are known to affect oocytes, but water hardness has not been considered an important factor in Xenopus husbandry by the research community. We found that, when females were kept in soft water or water with marine salts alone, even when it was cooled to 17 to 18 degrees C, the quality of oocytes decreased; only 20 to 25% of resulting embryos developed to tailbud stages. Survival and normal development of embryos increased significantly within one month of addition to the laboratory housing water of salts that mimic conditions in African Rift Valley lakes. These salts greatly increased water hardness; development of embryos to tailbud stages remained high (50 to 70% on average) for more than a year after their addition to the water housing females. Water from South African ponds where X. laevis are collected, and from wells used by the major suppliers of X. laevis, also was moderately to very hard. Our results suggest that X. laevis is naturally adapted to hard water, and indicate that increasing general hardness during laboratory housing is more important for oocyte quality and embryo development than is increasing carbonate hardness (alkalinity) in the water used to house females. PMID- 15134363 TI - Effect of sex and age on serum biochemical reference ranges in C57BL/6J mice. AB - The C57BL/6J mouse strain is widely used as a common genomic background for many gene-modified murine models. However, few data on its clinical biochemistry are available. Therefore, we conducted a study to provide new protocols for serum biochemical screening and developed the reference range for a set of 13 analytes that pertain to lipoprotein metabolism, electrolyte balance, and data reflecting function of the heart, liver, kidneys, and pancreas. Male and female mice were studied, and blood samples were obtained at six and 20 weeks of age. Of 13 parameters studied, 12 were affected by age and sex. Briefly, male mice had higher triglycerides, cholesterol, high-density lipoprotein cholesterol, glucose, and amylase values. With age, mice of both sexes developed higher triglycerides and glucose concentrations, as well as aspartate and alanine transaminase activities. A significant difference between mice and humans was noted for amylase activity, which is extremely high in this healthy mouse strain. Therefore, we suggest that caution should be taken when data are interpreted to indicate gastrointestinal disease in murine models. The reference values for murine clinical biochemical analytes obtained during the study reported here should be useful for characterizing the biochemical phenotype of experimental mice. PMID- 15134364 TI - Differences in spermatogenesis in cryptorchid testes among various strains of mice. AB - The purpose of the study reported here was to define strain differences in spermatogenesis in cryptorchid testes in mice. Mice of strains A/J, BALB/c, CBA/N, C3H/He, C57BL/6 (B6), ddY and ICR were found to be sensitive to heat stress attributable to experimentally induced cryptorchidism. In contrast, mice of strains AKR/N (AKR), MRL/MpJ-+/+ (M+) and MRL/MpJ-lpr/lpr (lpr) were resistant to heat stress. Relative increases of apoptotic cells were detected in the sensitive group, but not in the resistant group. A decrease of proliferating cell nuclear antigen-immunoreactive cells after experimentally induced cryptorchidism was observed only in the sensitive group. These results suggested that heat stress-resistant germ cells were present in MRL and AKR strains, possibly originating from the genetic background. PMID- 15134365 TI - Antimicrobial therapies for pulmonary Klebsiella pneumoniae infection in B6D2F1/J mice immunocompromised by use of sublethal irradiation. AB - Klebsiella pneumoniae is a common cause of nosocomially acquired pneumonia in immunocompromised patients. Previously, we established a pneumonia model using Klebsiella pneumoniae in B6D2F1/J mice sublethally irradiated with 7-Gy 60Co gamma-radiation and inoculated intratracheally. In the study reported here, we investigated survival of mice following 10 days of antimicrobial therapy with ceftriaxone, gentamicin, gatifloxacin, and a ceftriaxone-gentamicin combination given once daily. Survival was significantly prolonged in response to all therapies. However, survival of mice was 95% when treated with the ceftriaxone gentamicin combination followed by ceftriaxone alone (75%), and gatifloxacin (80%), whereas survival for controls was 0%. In addition, resistance to any of the treatments did not develop during the study. We conclude that an immunocompromised status does not alter the Infectious Disease Society of America's primary recommendation for treating community-acquired K. pneumoniae pneumonia using a third-generation cephalosporin, with or without an aminoglycoside. PMID- 15134366 TI - Effects of geographic origin on captive Macaca mulatta mitochondrial DNA variation. AB - Partial sequences from mitochondrial (mt) 12S and 16S rRNA genes were analyzed to characterize diversity among captive rhesus macaques (Macaca mulatta) originating from various geographic regions. Several nested clades, defined by closely related haplotypes, were identified, suggesting considerable genetic subdivision, probably relics from heterogeneous origins, founder effects, and genetic drift, followed by breeding isolation. The rhesus matrilineages from India differed discretely and markedly from Chinese matrilineages; approximately 90% of the genetic heterogeneity among the combined samples of Indian and Chinese rhesus macaques studied here was due to country of origin. In addition, mtDNA sequences from macaques of China were more diverse than those from rhesus macaques of India, an outcome consistent with China's greater subspecies diversity and with nuclear genotype distributions. Otherwise, the distribution of mtDNA variation within rhesus macaques of China, and especially within those of India, exhibited far less structure and did not conform to a simple isolation-by-distance model. As the demand for genetically heterogeneous and well-characterized rhesus macaques for biomedical-based research increases, mtDNA haplotypes can be useful for genetically defining, preserving maximal levels of genetic diversity within, and confirming the geographic origin of captive breeding groups of rhesus macaques. PMID- 15134367 TI - Neonatal anesthesia for studies of hamster parental behavior when infanticidal aggression is a possibility. AB - Experiments involving investigation of the neuroendocrine basis for paternal care in rodents risk activation of aggressive behavior toward pups. To minimize pain and suffering during tests of parental responsiveness requiring retrieval of a displaced pup to its nest, a method of anesthetizing the pup was developed in Djungarian hamsters, Phodopus campbelli. A surgical plane of anesthesia, as measured by criteria, such as respiratory depression, loss of the pedal reflex, and failure to increase respiratory rate or to vocalize in response to handling, was achieved by use of intraperitoneal administration of a combination of ketamine and xylazine. Both parents (tested separately) expressed normal behavior toward anesthetized pups. In random order, a saline-injected or anesthetized pup was displaced from its nest in the home cage. There were no differences in pick up or retrieval rates between saline and anesthetized pups for either parent. A third test using an unmanipulated pup confirmed that parental behavior was not reduced toward an anesthetized pup. However, if anesthetized pups were tested first among littermates, retrieval by males was less likely. This method will, therefore, underestimate retrieval behavior in males, but not females. Adult male hamsters that had never been parents also expressed expected behavior by attacking the pup in 45% of cases. This method provides an efficient and effective means of protecting pups while allowing adults to express a wide range of parental and infanticidal behaviors. It also has application in behavioral screening of transgenic strains toward unrelated young. PMID- 15134368 TI - Model of angiogenesis in mice with severe combined immunodeficiency (SCID) and xenoengrafted with Epstein-Barr virus-transformed B cells. AB - Xenoengraftment of human cells in mice with severe combined immunodeficiency (SCID) has been used as a model system to study the mechanisms of B-cell lymphomagenesis. In the study reported here, we determined that SCID mice can also be used as a model to study angiogenesis in B-cell lymphomas. The C.B-17 scid/scid mice were xenotransplanted with Epstein-Barr virus (EBV)-transformed lymphoblastoid cell lines (LCL), and we determined whether CD31, a marker found on endothelial cells, was detected in the human B-cell lymphomas that developed in these mice. Microvessel formation was identified by use of immunohistochemical staining for CD31. To assess possible mechanisms of angiogenic stimulus, we analyzed the expression of interleukin 8 (IL-8), a chemokine documented to promote angiogenesis, in non-small-cell lung cancer and bronchogenic carcinomas. We observed that a panel of LCL and LCL-lymphomas expressed IL-8 mRNA and protein. Neutralization of IL-8, however, did not inhibit lymphomagenesis, suggesting that IL-8 is not essential for angiogenesis in this model. To examine other parameters of angiogenesis, we identified expression of vascular endothelial growth factor in the lymphomas. These data suggest that angiogenesis accompanies EBV-associated B-cell lymphoma development, but IL-8 is not essential for this process. Thus, the SCID mouse model is amenable to testing of anti angiogenic factors. PMID- 15134369 TI - Barbering (fur and whisker trimming) by laboratory mice as a model of human trichotillomania and obsessive-compulsive spectrum disorders. AB - Animal diseases that develop spontaneously in a limited subpopulation can provide powerful models of human disease because they provide a means to investigate the interaction of a broad range of biological and environmental etiologic processes. In contrast, with experimentally induced animal models, the etiology of the model is inherently fixed, and can only speak to a limited subset of those involved in the human disease. 'Barbering' (abnormal whisker- and fur-plucking behavior) in mice resembles human trichotillomania (compulsive hair plucking) in that barbering mice pluck focused areas of hair, and engage in post-plucking manipulatory and oral behaviors. We performed a cross-sectional epidemiologic survey of a population of 2,950 laboratory mice to further assess the face validity of barbering as a spontaneous model of trichotillomania. Patterns of hair loss and demographic and etiologic risk factors were recorded for each mouse, and were analyzed by use of logistic regression. Barbering paralleled trichotillomania in terms of phenomenology, demography, and etiology. Thus, similar to trichotillomania, barbers predominately plucked hair from the scalp and around the eyes and the genitals; barbering was female biased, and had its onset during puberty; and etiologic factors included reproductive status and genetic background. Therefore, barbering has excellent face validity as a model of trichotillomania, and may represent a refined and non-invasive model, especially for studies of the complex genetic/environmental etiologies of this disorder. PMID- 15134371 TI - The relationship between the pleasure of wine tasting and its beneficial effects. PMID- 15134370 TI - Wine and health. PMID- 15134372 TI - Overview of epidemiological studies on wine, health and mortality. AB - Numerous epidemiological studies have observed that moderate intake of alcohol including wine is associated with a lower risk of cardiovascular disease (CVD). However, according to several authors, moderate consumption of wine is more beneficial than that of beer or spirits. Some studies have shown that moderate consumption of wine can lower mortality from CVD and other causes. The link between drinking wine and total mortality risk (all causes combined) has been studied. The results of various prospective population studies show that intake of beer and spirits from abstention to light to moderate daily intake did not influence mortality, while wine seems to have a beneficial effect on all causes of mortality. Other studies have reached the same conclusion. In general, several authors have reported that in subjects consuming wine in moderation the risk of mortality from all causes is 20-30% lower than in abstainers. Grape wine appears to be the main alcoholic beverage that contains antioxidant phenolic substances known to inhibit oxidation of low-density lipoprotein and affect hemostasis and carcinogenesis. In conclusion, there are differences in the effects of wine, beer and spirits on health. These differences may not be significant in coronary heart disease. Only moderate wine consumption appears to have a beneficial effect on several types of cancer and on total mortality. PMID- 15134373 TI - Role of viticultural factors on stilbene concentrations of grapes and wine. AB - Stilbenes are phenolics that can be found in different grapevine organs, such as berries, leaves, canes and roots. Stilbenes act as antifungal compounds since they are synthesized by the plant in response to pathogen attack. Other abiotic elicitors, such as ultraviolet rays and heavy metals, can trigger stilbene production. Many stilbenic compounds have been detected in grapes and wine (resveratrol, piceid, viniferins, astringin, etc.). The latest stilbene to be analyzed is piceatannol (or astringinin). Stilbene synthesis in grapes depends on different viticultural factors such as the grape variety, the environment and cultural practices. Concerning grape variety, red berry-grapes have higher stilbene levels than white berry-grapes. With regard to climate, preliminary results suggest a positive correlation between vineyard elevation and stilbene grape concentrations. Quality-oriented cultural practices produce grapes with high levels of stilbenes. PMID- 15134374 TI - Antioxidants in Sicilian wines: analytic and compositive aspects. AB - The beneficial effects of wine are associated with the physiological protection conferred by phenolic compounds such as anthocyanins and resveratrol. Levels of these phenolic compounds were quantified in 19 monovarietal wines produced in Sicily. Resveratrol and resveratrol-glucosides were detected by high-performance liquid chromatography (HPLC) with an ultraviolet detector, while anthocyanins were determined by micro-HPLC-Electron Spray Ionization-Mass Spectroscopy (ESI MS) analysis. The amount of cis- and trans-resveratrol and of cis- and trans piceid varied in the different types of wine, depending on the grape variety. Red wines presented higher contents of resveratrol and resveratrol-glucosides, whereas lower concentrations were present in white wines. In Merlot wine, the concentration of trans-piceid (5.04 mg/l) was significantly greater than in the other wines and represented the highest concentration among all the resveratrol isomers. Fourteen components were identified and dosed in the anthocyanin fraction. The highest concentration of total anthocyanins (417 mg/l) was found in the Cabernet Sauvignon wine, while the highest value among the wines made from the autochthonous grapes was found in Nero d'Avola. Antioxidant capacity was also studied. The results show that the antioxidant capacity of wines is strictly related to the amount of phenolic compounds. PMID- 15134375 TI - Bioavailability of tyrosol, an antioxidant phenolic compound present in wine and olive oil, in humans. AB - Tyrosol is a phenolic compound present in two of the traditional components of the Mediterranean diet: wine and virgin olive oil. The presence of tyrosol has been described in red and white wines. Tyrosol is also present in vermouth and beer. Tyrosol has been shown to be able to exert antioxidant activity in in vitro studies. Oxidation of low-density lipoprotein (LDL) appears to occur predominantly in arterial intima in microdomains sequestered from antioxidants of plasma. The antioxidant content of the LDL particle is critical for its protection. Thus, phenolics, which are able to bind LDL, could be effective in preventing lipid peroxidation and atherosclerotic processes. The ability of tyrosol to bind human LDL has been reported. We have demonstrated the bioavailability of tyrosol in humans from virgin olive oil in its natural form. Urinary tyrosol increased, reaching a peak at 0-4 h after virgin olive oil administration. Men and women showed a different pattern of urinary excretion of tyrosol. Moreover, tyrosol is absorbed in a dose-dependent manner after sustained and moderate doses of virgin olive oil. In summary, our results suggest that tyrosol from wine or virgin olive oil could exert beneficial effects on human health in vivo if its biological properties are confirmed in in vivo studies. PMID- 15134376 TI - Procyanidins from Vitis vinifera seeds display cardioprotection in an experimental model of ischemia-reperfusion damage. AB - Since the early 1970s, increasing evidence has suggested that the consumption of moderate amounts of alcohol is inversely correlated with mortality from myocardial infarction. There is also some evidence that the protective effects of wine might be more pronounced than those of other alcoholic beverages. These observations prompted us to investigate the cardioprotective activity of Vitis vinifera seeds in experimental ischemia-reperfusion injury. An isolated rabbit heart preparation paced electrically was used to evaluate the effects of a highly purified, high molecular weight fraction of oligomeric procyanidins isolated from Vitis vinifera seeds on myocardial reperfusion injury after 40 min of low-flow (1 ml/min) ischemia. Infusion of the heart with 100 or 200 microg/ml procyanidins dose-dependently reduced left ventricular end-diastolic pressure during ischemia, decreased coronary perfusion pressure, improved cardiac mechanical performance upon reperfusion, increased the release of 6-Keto-prostaglandin F1alpha into the perfusate in both the preischemic and the reperfusion periods and suppressed rhythm irregularity. Procyanidins dose-dependently relaxed human internal mammary aortic (IMA) rings (with intact endothelium) precontracted with norepinephrine. This effect was completely abolished in IMA-rings without functional endothelium or when this vascular tissue was pretreated with nitric oxide synthase inhibitor (NG-monomethyl-L-arginine) or with guanylate cyclase inhibitor (1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one). In conclusion, these results indicate that procyanidins could be of therapeutical potential in cardiovascular diseases. However, further investigations are required for a better definition of the mode of action of these oligomers. PMID- 15134377 TI - Postprandial effects of wine consumption on lipids and oxidative stress biomarkers. AB - Postprandial lipemia has been recognized as a risk factor for atherosclerosis development. Consuming meals with suitable sources of antioxidants such as red wine reduces postprandial oxidative stress. However, information about the postprandial effects of wine ingestion outside meals on lipids and on in vivo low density lipoprotein (LDL) oxidation in humans is scarce. The aim of this study was to investigate postprandial changes in lipids and in vivo LDL oxidation after moderate (250 ml) red wine ingestion, before and after sustained wine consumption of 250 ml/day for 4 days. After 4 days of sustained wine consumption a decrease in the LDL/high-density lipoprotein cholesterol ratio was observed after wine ingestion (p = 0.026). On day 4, a decrease in oxidized LDL levels and an increase in the antioxidant enzyme glutathione peroxidase activity (p = 0.025) were observed after wine ingestion. Our results show that consumption of red wine at moderate doses outside meals does not promote oxidative stress. Daily consumption of moderate doses of red wine can improve postprandial lipid profile and oxidative status when wine is ingested outside meals. PMID- 15134378 TI - Wine in the prevention of chronic bacterial urinary infection. AB - The observation of statistical public health data, together with lifestyle in a still native population following a strict Mediterranean diet, where local wine consumption of approximately 350 ml daily still has a primary role, demonstrates that this area, Pantelleria, also called the Black Pearl, has better regulation of common intestinal motor disorders. The incidence of chronic bacterial urinary infection is 30% lower than the national average. Further open laboratory studies should be performed to confirm our data and to elucidate whether protection against chronic bacterial urinary infection is congenital or acquired. PMID- 15134379 TI - Evidence for resveratrol-induced preservation of brain mitochondria functions after hypoxia-reoxygenation. AB - We have previously shown, as have other authors, that trans-resveratrol (E resveratrol, 3,4,5-trihydroxy-E-stilbene) reduces reactive oxygen species (ROS) generation of mitochondria freshly isolated from healthy rat brains and that it also counteracts the effect of uncouplers (CCCP) on mitochondrial respiration and oxidative phosphorylation. Two main mechanisms have been shown: firstly, a scavenger effect toward O2- and secondly inhibition of complex III ROS generation. We now report on the effects of resveratrol in a pathological model that mimics the ischemia followed by the reperfusion process which may occur in the human brain. Isolated brain mitochondria were submitted first to hypoxia then to reoxygenation. The aim of this study was to determine the extent of mitochondrial damage induced by this experimental model, to demonstrate which mitochondrial functions were altered and to quantify the extent to which they were prevented by resveratrol. Resveratrol was either added to mitochondria freshly isolated from healthy rat brains or was injected by subcutaneous chronically implanted pumps (0.5, 2 and 10 mg/kg/day for 7 days). The rats were then sacrificed and mitochondria were extracted from brains. To evaluate the respective effects of hypoxia and reoxygenation on mitochondrial functions and the relevant effects of resveratrol, this drug was added (first protocol) either before the complete process (i.e., hypoxia and reoxygenation), or after anoxia before reoxygenation. We found that resveratrol prevented alterations of mitochondrial functions. This substance partly counteracted the decrease in respiratory control and the increase in ROS generation. It fully inhibited the alteration of membrane fluidity and the mitochondrial step of the apoptotic process (evidenced by cytochrome c release and membrane potential collapse). The effects of resveratrol were concentration-dependent (in vitro) or dose-dependent (ex vivo, second protocol). They were not significantly different when the drug was added before or after hypoxia, which suggests that in this model, reoxygenation was the most deleterious process and the stage at which resveratrol was most effective. PMID- 15134380 TI - Wine and endothelial function. AB - In recent years many studies have focused on the well-known relationship between wine consumption and cardiovascular risk. Wine exerts its protective effects through various changes in lipoprotein profile, coagulation and fibrinolytic cascades, platelet aggregation, oxidative mechanisms and endothelial function. The last has earned more attention for its implications in atherogenesis. Endothelium regulates vascular tone by a delicate balancing among vasorelaxing (nitric oxide [NO]) and vasoconstrincting (endothelins) factors produced by endothelium in response to various stimuli. In rat models, wine and other grape derivatives exerted an endothelium-dependent vasorelaxing capacity especially associated with the NO-stimulating activity of their polyphenol components. In experimental conditions, reservatrol (a stilbene polyphenol) protected hearts and kidneys from ischemia-reperfusion injury through antioxidant activity and upregulation of NO production. Wine polyphenols are also able to induce the expression of genes involved in the NO pathway within the arterial wall. The effects of wine on endothelial function in humans are not yet clearly understood. A favorable action of red wine or dealcoholized wine extract or purple grape juice on endothelial function has been observed by several authors, but discrimination between ethanol and polyphenol effects is controversial. It is, however likely that regular and prolonged moderate wine drinking positively affects endothelial function. The beneficial effects of wine on cardiovascular health are greater if wine is associated with a healthy diet. The most recent nutritional and epidemiologic studies show that the ideal diet closely resembles the Mediterranean diet. PMID- 15134381 TI - Effect of resveratrol and catechin on PC12 tyrosine kinase activities and their synergistic protection from beta-amyloid toxicity. AB - beta-Amyloid peptide (beta-AP) is the main component of amyloid deposits around the cerebral vessel and in the brain parenchyma in Alzheimer's disease and Down's syndrome. In vitro studies in neuronal cells or in PC12 and Hela cell lines have shown that the aggregate form of beta-AP is toxic. Many genetic and environmental factors including metal ions, proteoglycans, plasma proteins and antioxidants modify beta-AP toxicity. We investigated the effect of two plant polyphenols- resveratrol and catechin--on soluble and particulate tyrosine kinase activity from PC12 cells and the protective action of these compounds against beta-AP (1 41) toxicity. beta-AP (1-41) decreased PC12 viability with an IC50 value of 1.1 +/- 0.14 x 10(-8) M. Resveratrol and catechin protected PC12 cells from beta-AP (1-41) toxicity. With 25 microM resveratrol the IC50 value increased to 2.2 +/- 0.19 x 10(-7) M. In the presence of beta-AP (1-41) resveratrol showed a concentration-dependent biphasic effect, and at a concentration of up to 40 microM it protected PC12 cells from beta-AP (1-41) toxicity. At concentrations higher than 40 microM, an inhibitory activity on cell proliferation appeared. This antiproliferative effect was also seen in the absence of beta-AP (1-41). With 100 microM catechin the IC50 value increased from 1.1 +/- 0.14 x 10(-8) M to 3.2 +/- 0.25 x 10(-7) M beta-AP (1-41). The protective effect was concentration dependent. Resveratrol and catechin had a synergistic protective action. In the presence of 40 microM catechin and 10 microM resveratrol or 20 microM resveratrol and 10 microM catechin, the toxicity determined by 10(-7) M beta-AP (1-41) was almost completely removed. Resveratrol and catechin had different effects on PC12 tyrosine kinase activity. With peptide 1-17 of gastrin as substrate, resveratrol inhibited particulate tyrosine kinases while it had no effect on soluble activity. With the same substrate, catechin increased the activity of soluble fraction while it inhibited particulate activity. When peptide 6-20 of cell division kinase p34cdc2 was utilized, catechin showed an opposite effect, inhibiting soluble tyrosine kinase activity and increasing particulate activity. With peptide 6-20, resveratrol inhibited both soluble and particulate activities. These results demonstrate that resveratrol and catechin have different activities on the signal transduction pathway involving protein phosphorylation. These differences may contribute not only to the different effects of these compounds on PC12 growth but also to the synergistic effect against beta-AP (1-41) toxicity. The different activity of resveratrol and catechin on signal transduction pathways, as well as the differences in metal chelation, partition coefficient between water and lipids, hydrogen donation redox potential and enzyme inhibition may be at least in part based on synergistic protection against beta-AP (1-41) toxicity. PMID- 15134382 TI - Wine and tumors: study of resveratrol. AB - In modern industrial societies the attention to public health, especially in relation to food habits, is increasing day by day. Considering this, it's no wonder that wine, the voluptuary drink that best represents human history, is the most interesting compound. The main and best known wine effects on the human body are caused by alcohol, but several other active compounds are present in wine. Above all, resveratrol is able to neutralize free radicals, which can damage DNA and may lead to cancer onset. In this study, we have indagated resveratrol anticancer action, analyzing its effects on both cell cycle and growing of human lymphoma B (DHL-4) cells. MTT colorimetric test, tripan blue dye exclusion assay, and cell cycle analysis showed that resveratrol has a dose-dependent antiproliferative and antiapoptotic action on DHL-4 cells. These results confirm resveratrol's potential therapeutic role on tumors. PMID- 15134383 TI - Resveratrol structure and ceramide-associated growth inhibition in prostate cancer cells. AB - Resveratrol (3,4',5-trans-trihydroxystilbene) is a dietary polyphenol with chemopreventive properties present in grapes, red wine, peanuts and other edible products. The antiproliferative and proapoptotic effect of resveratrol in breast cancer cells can be traced to the accumulation of ceramide. In this study we demonstrate that resveratrol can also exert antiproliferative/proapoptotic effects in association with the accumulation of endogenous ceramide in the androgen receptor (AR)-negative prostate cancer cell line, PC3. Notably, resveratrol shares with other ceramide-inducing agents a phenolic moiety on its structure. For this reason we hypothesize that the phenolic moiety is critical for the ceramide-associated growth-inhibitory effects of resveratrol. We compared the ability to induce both ceramide increase and growth inhibition in PC3 cells of resveratrol and three resveratrol analogs: piceatannol (3,3',4',5-trans tetrahydroxystilbene), with an additional hydroxyl group in the 3' position; trans-stilbene, the nonhydroxylated analog; and the semisynthetic 3,4',5 trimethoxy-trans-stilbene (TmS), with methoxyl groups in lieu of the hydroxyl groups. Of the three stilbenoids, only piceatannol (and not stilbene or TmS) produced ceramide-associated growth inhibition. These data point to the phenolic moiety of stilbenoids as a critical structural feature necessary to induce ceramide-associated growth inhibition. PMID- 15134384 TI - Relationship between iron and protein content of dishes and polyphenol content in accompanying wines. AB - The traditional combination of wines and dishes is highly complex, elaborated and refined. The aim of this study was to investigate the possible relationship between the chemical composition of wines and dishes that determines their combination. We determined the content of total polyphenols in 56 wines. The content of total proteins, total lipids, kilocalories, sodium, potassium, calcium, copper and zinc were determined in 44 raw foods and 44 dishes. Nine gourmets independently chose three wines for each food. We correlated the content of the chemical constituents of foods with the phenol content of wines combined with each food by the gourmets. A significant positive correlation was obtained between the phenol content of wines and iron (r = 0.81, p < 0.0001) and total protein content (r = 0.66, p < 0.0001) of foods. Nine gourmets composing a second panel chose three wines for each dish. A significant positive correlation was also obtained between the phenol content of wines and iron (r = 0.69, p < 0.0001), total protein (r = 0.50, p < 0.0006) and potassium (r = 0.45, p < 0.002) in dishes combined with wines by the second panel of gourmets. Plant phenols decrease the intestinal absorption of iron and have antioxidant activity in the intestinal tract and elsewhere in the body. These positive effects compensate the negative antinutritional activity toward protein digestion. The traditional combination of wines and dishes appears to be very favorable since wines poor in phenols are combined with dishes poor in iron and/or proteins to minimize their possible antinutritional effects, while phenol-rich wines are combined with dishes rich in iron to decrease iron absorption and prandial peroxidative stress. PMID- 15134385 TI - Percutaneous absorption of flavan-3-ol conjugates from plant procyanidins. AB - Catechins (flavanols) are strong antioxidants, free radical scavengers and inhibitors of lipid peroxidation. New bio-based antioxidant compounds obtained by depolymerization of plant polymeric flavanols (procyanidins) in the presence of cysteine or cysteamine, as well as their underivatized counterpart, (-) epicatechin, were evaluated in terms of their percutaneous absorption profiles taking into account their free radical scavenging efficiency. The evaluation of the percutaneous absorption of flavanols was carried out by an in vitro methodology using both pig and human skin. A good correlation was obtained using both skins in the evaluation of the skin absorption profiles. It can be deduced that 4beta-(S-cysteinyl)epicatechin (Cys-Ec) has a tendency to be located mainly in the outermost layers of the skin, whereas 4beta-(2-aminoethylthio)epicatechin (Cya-Ec) has a pronounced percutaneous absorption capacity. Their antioxidant properties and their skin penetration profiles support their potential cosmetic or pharmacological applications. PMID- 15134386 TI - A two-year study of chondroitin sulfate in erosive osteoarthritis of the hands: behavior of erosions, osteophytes, pain and hand dysfunction. AB - The aim of this study was to evaluate the effect of 800 mg/die of chondroitin sulfate (CHS) per os plus naproxen versus naproxen over 2 years in patients with erosive osteoarthritis (EOA) of the hands. Joint count for erosions, Heberden and Bouchard nodes, Dreiser's algofunctional index and physicians' and patients' global assessment of disease activity were studied. A total of 24 consecutive patients (22 women and 2 men, mean age 53.0 +/- 6) suffering from symptomatic OA with radiographic characteristics of EOA were evaluated. The patients were divided into two groups of 12 patients each. The first group took naproxen 500 mg only. The second group was treated with CHS 800 mg orally plus naproxen 500 mg. Joint counts, radiological hand examinations and assessment of disease activity were performed at baseline, at 12 months and at 24 months. In the second year the treated group showed significant worsening in erosion, Heberden, Bouchard and Dreiser scores was recorded. Physician and patient global assessments of disease activity showed no significant difference from baseline scores. The untreated group showed significant worsening in erosion, Heberden and Bouchard nodes, Dreiser index and physician and patient global assessment scores. This study confirms the partial efficacy of oral CHS in improving some aspects of EOA. PMID- 15134387 TI - Evaluation of the degree of susceptibility of Streptococcus pyogenes erythromycin resistant strains to rokitamycin (a 16-membered macrolide) using the Epsilometer test. AB - Routine hospital screening of the resistance of Streptococcus pyogenes to macrolides is usually done using the erythromycin, clarithromycin or azithromycin disk diffusion technique. When a strain is found to be resistant to one of these macrolides, it is generally assumed to be resistant to the whole class. However this approach gives only partial qualitative information because S. pyogenes strains with inducible and M phenotype resistance are still susceptible to 16 membered ring macrolides such as rokitamycin. Seventy-four erythromycin-resistant (22 inducible and 52 M phenotype) strains of S. pyogenes were tested for their susceptibility to rokitamycin and clindamycin (control) by means of the agar disk diffusion test and the results were compared with those obtained using the Epsilometer test, a quantitative technique for measuring bacterial susceptibility and minimal inhibitory concentrations (MIC). Epsilometer testing of erythromycin in comparison with rokitamycin is useful for measuring the real degree of susceptibility of macrolide-resistant strains quickly and simply. This is important because strains with the same disk diffusion diameter do not necessarily have the same MIC, but a scattered distribution of susceptibility. PMID- 15134388 TI - Treatment of Alzheimer's disease with stabilized oral nicotinamide adenine dinucleotide: a randomized, double-blind study. AB - This study was designed to evaluate the effect of stabilized oral reduced nicotinamide adenine dinucleotide (NADH) on cognitive functioning in patients with Alzheimer's disease (AD). NADH is a coenzyme that plays a key role in cellular energy production and stimulates dopamine production. In previous trials NADH has been shown to improve cognitive functioning in patients with Parkinson's disease, depression and AD. The present trial was a randomized, placebo controlled, matched-pairs, double-blind, 6-month clinical study. Patients with probable AD (n = 26) were randomized to receive either stabilized oral NADH (10 mg/day) or placebo. Twelve pairs of subjects were matched for age and baseline total score on the Mattis Dementia Rating Scale (MDRS) and the Mini Mental State Examination. After 6 months of treatment, subjects treated with NADH showed no evidence of progressive cognitive deterioration and had significantly higher total scores on the MDRS compared with subjects treated with placebo (p < 0.05). Analysis of MDRS subscales revealed significantly better performance by NADH subjects on measures of verbal fluency (p = 0.019), visual-constructional ability (p = 0.038) and a trend (p = 0.08) to better performance on a measure of abstract verbal reasoning. There were no differences between groups in measures of attention, memory, or in clinician ratings of dementia severity (Clinical Dementia Rating). Consistent with earlier studies, the present findings support NADH as a treatment for AD. PMID- 15134389 TI - Effects of D-003, a mixture of high molecular weight aliphatic acids from sugar cane wax, on bones from ovariectomized rats. AB - Bisphosphonates inhibit bone resorption through mechanisms involving the metabolic pathway from mevalonate to cholesterol. Mevalonate is a precursor of the lipoids required for osteoclast activity and thus inhibition of its synthesis affects bone metabolism. Inhibitors of hydroxymethylglutaryl CoA (HMGCoA) reductase might increase experimentally new bone formation through a mevalonate dependent effect. D-003 is a mixture of high molecular weight fatty acids isolated from sugar cane wax, which inhibits cholesterol biosynthesis through indirect regulation of HMGCoA reductase activity. This study was undertaken to determine whether D-003 could prevent bone loss in ovariectomized rats. Sprague Dawley female rats were ovariectomized or sham operated and were randomly distributed into five groups: a control ovariectomized and a sham (false operated) group receiving Tween/H2O vehicle, a group treated with alendronate (3 mg/kg/day) and two groups treated with D-003 (50 and 200 mg/kg/day). Treatments were administered for 3 months. At sacrifice, bones were removed and histological variables of bone resorption and formation were studied by histomorphometry. Ovariectomy diminished trabecular number and thickness and increased trabecular gap, osteoclast number and surface. Alendronate and D-003 prevented a decrease in trabecular number and thickness as well as increases in trabecular separation, osteoclast number and surface compared with ovariectomized controls, thus preventing the bone loss and decreased bone resorption induced by ovariectomy, but failed to increase osteoblast surface compared with control ovariectomized rats. In conclusion, D-003 (50 and 200 mg/kg/day) prevented bone loss and decreased bone resorption in ovariectomized rats, which suggests that this substance could be promising in preventing or treating osteoporosis. PMID- 15134390 TI - Linear arrays for vibro-acoustography: a numerical simulation study. AB - In this work, we study linear array beamforming for vibro-acoustography systems. The goal of this research is to assess the feasibility of vibro-acoustography imaging systems based on linear arrays for clinical applications. The vibro acoustography transducer is designed with two arrays driven by two continuous wave or tone-burst signals at slightly different frequencies. The system beamforming is modeled in a homogeneous and lossless fluid. In this model, the point-spread function (psf) of the system is obtained as the product of the spatial impulse function of each linear array. Three linear array configurations are analyzed for vibro-acoustography systems. Simulations considering 64 elements in each array are performed. Aspects related to clinical applications ofvibro acoustography, such as system spatial resolution, sidelobes, grating lobes and image frame-rate are discussed. It is concluded that linear array transducers can produce acceptable spatial resolution for clinical applications. PMID- 15134391 TI - Elastographic imaging of thermal lesions in liver in-vivo using diaphragmatic stimuli. AB - Radiofrequency or microwave ablations are interstitial focal ablative therapies that can be used in a percutaneous fashion for treating tumors in the liver, kidney, and prostate. These modalities provide in situ destruction of tumors. We present a method for in-vivo elastographic visualization of the ablated regions in the liver during and after thermal therapy. In-vivo elastographic imaging uses compressions of the liver due to movement of the diaphragm during the respiratory cycle. Elastography of the liver and other abdominal organs has not been attempted previously due to the difficulty in providing controlled compressions. Gating of the data acquisition to the respiratory waveform would provide access to data where the compression increments are similar in both magnitude and direction, thereby enabling reproducible imaging of the thermal lesion or tumor. Comparison of elastograms with gross-pathology of ablated tissue illustrates the correspondence between elastographic image features and pathology. Ultrasound is routinely used to guide the rf ablation procedure, so the same imaging system could be used for elastographic imaging. Since the technique utilizes physiological motion of the diaphragm due to respiration, it may also be employed in the visualization of cancerous tumors in the liver. PMID- 15134392 TI - A novel and robust method for rapid strain estimation in elastography. AB - In elastography, change in signal shape from tissue deformation and nonaxial tissue motion reduce correlation between the pre- and postcompression echo signals. Appropriate global temporal stretching of postcompression signals can reduce the decorrelation. Adaptive stretching performs a search for the stretch factor that maximizes the correlation between the pre- and postcompression echo signal segments at each data window location. Adaptive stretching is robust but computation intensive. In contrast, global stretching is fast but performs well only in areas where local strains are close to the applied strain. We developed a method that strikes a balance between the speed of global stretching and the performance of adaptive stretching. In this method, several strain maps are computed by performing global stretching with a range of different stretch factors. The area in each computed strain image with strain values closely corresponding to the uniform stretch factor will contain 'good quality' strain estimates. To produce a single elastogram at the end, we identify the strain map with the maximum correlation at each location and the strain value in that strain map at that location is chosen for the combined map. Results from data generated by finite-element simulation and phantom experiments demonstrate that the described strain estimator is significantly less susceptible to signal degradation than conventional strain estimators. PMID- 15134393 TI - Metanalytical assessment of reference values for polychlorinated biphenyl in human blood. AB - Due to their physico-chemical characteristics, polychlorinated biphenyls (PCBs) are highly persistent in the environment and therefore easily measured in the biological matrices of more and more large groups of general population. For these reasons it would be useful to determine suitable Reference Values (RVs) for these xenobiotics. In this paper, a metanalysis to the published data on PCBs values in human blood is presented. This investigation was carried out in order to reach adequate information on their RVs and to focus some specific topics to be taken into account when producing directly RVs for PCBs. The PCBs RVs resulted between 1.2 e 8.28 microg/L for males and between 2.69 e 5.17 microg/L for females the general range varied from 0.9 to 56 microg/L. The main criticisms in the assessment of RVs for PCBs resulted: the number of examined subjects; the inclusion and stratification criteria; the analytical method adopted and their quality assurance; the type and number of congeners to be determined and their specific quantification; the calculation of blood PCBs concentration (weight/volume or weight/lipids); the statistical analysis and in particular the treatment of not detectable data. PMID- 15134394 TI - [Survey on educational needs in occupational medicine in Italy]. AB - The importance to correctly address continuing education among occupational health professionals in Italy has been particular in the focus of the Scientific and Professional Medical Community also in consideration of the recent regulations implemented in the field of Continuing Medical Education (CME). The Italian Society of Occupational Medicine recently developed an advanced programme of Continuing Medical Education for Occupational Physician. In order to correctly asses the education needs and collect valid information of Occupational Physicians in Italy we administered a target questionnaire among a group of them. The results are analyzed and discussed. PMID- 15134395 TI - [Evaluation of the levels of radiofrequency electromagnetic fields in the territory of the city of Bari in outside and inside environments]. AB - In this study we measured the levels of the high frequency field in the proximity of non-ionizing radiation sources (wireless transmitting stations for mobile telephones and radio and television transmitters) in nine districts of the city of Bari. The measurements were taken both inside and outside closed environments. For the indoor measurements we took into account electromagnetic field generating equipment (VDT, electric domestic appliances, mobile telephones) in working and non-working order and with the windows open and shut respectively. We carried out these measurements according to the methods laid down in the Italian regulation CEI ENV 50166-2 of May 1995, as shown in the enclosure to the Ministerial Decree of 10.9.98 n.381. The electromagnetic field levels near wireless transmitting stations for mobile telephones are certainly modest when we consider that they never exceeded the limits established by the aforesaid Ministerial Decree. On the contrary radio and television equipment creates a much greater source of exposure. The electromagnetic field levels are certainly superior to those of the wireless transmitting stations although they never exceed, except in one isolated case, the values established by the Ministerial Decree 381/98. PMID- 15134396 TI - [Emergent pathology in occupational medicine: the mobbing]. AB - Mobbing constitutes a phenomenon not yet clearly defined. The activity, located in the II Faculty of Medicine and Surgery, is specifically dedicated to such phenomenon, which is the outcome of cooperation. The examined population is composed by a light prevalence of men (51.3%) respect to women (48.6%), aged around 42 years and employees (71.6%), workers (28.4%). All patients have been submitted to working anamnesis before being subjected to a psychiatric examination and psycodiagnostic tests. The emerged data underline a trouble of adaptation 55.4% of the cases, 40.5% is affected by psychiatric pathologies, 4.1% of the patients do not show mental disorders. A certification of compatibility with mobbing has been possible to be verified in 49% of the cases. Nevertheless, it is necessary a connection between a physician and business medical service. We hope a necessary enactment of a specific regulation. PMID- 15134397 TI - [Sleep-disordered breathing and occupational medicine: considerations on three clinical cases]. AB - Three subjects suffering from sleep-disordered breathing are described. Two of them (a 37-year-old lorry driver with obstructive sleep apnea, and a 47-year-old bricklayer with upper airway resistance syndrome) had suffered several accidents at work in few months, due to daytime sleepiness. The third patient (45-year-old) had developed (central and obstructive) sleep apnea after having been exposed to organic solvents for 15 years as a painter. In all the three subjects, polysomnography led to correct diagnosis, and nocturnal ventilotherapy with nasal CPAP (continuous positive airway pressure) considerably improved the clinical picture. The cases presented illustrate the twofold interest of sleep-disordered breathing for Occupational Medicine: on the one hand, these syndromes may reduce job performance and increase the accident risk; on the other hand, they may recognize as a risk factor the occupational exposure to solvents. PMID- 15134398 TI - [Report of courses on "Occupational dermatology" and "Occupational skin and respiratory allergies" organized by the Nordic Institute for Advanced Training in Occupational Health (NIVA)]. AB - The Nordic Institute of Advanced Training in Occupational Health (NIVA), has been organising courses for around 10 years, aimed to the constant up-date of Occupational Medicine. The courses "Occupational dermatology" and "Occupational skin and respiratory allergies", held in 2001 and 2002, analysed some occupational medicine aspects such as allergic contact dermatitis in metal workers, latex disease, allergies in odontoiatric workers, correct patch test performing. The courses has underlined the importance of the cooperation between occupational physician and dermatologist or other specialists, and the project of a standard questionnaire monitoring the exposure to allergic substances. This article is a summary of an extended publication available on the following URLs: http://www.unibs.it/medlav http://www.gimle.fsm.it. PMID- 15134399 TI - [Occupational exposure in the textile sector]. PMID- 15134400 TI - [Data on occupational activities in the textile sector]. PMID- 15134401 TI - [Spinning of carded textiles]. PMID- 15134402 TI - [Safety in the textile compartment -- risk profile "Manufacture of items of clothing"]. PMID- 15134403 TI - [Dying and printing of textiles]. PMID- 15134404 TI - [Occupational allergic contact dermatitis caused by dimethylmelamine resin in a textile worker]. PMID- 15134405 TI - [Assessment of the chemical risk in wool textile plant in Piedmont]. PMID- 15134407 TI - Multidisciplinary is not a dirty word. PMID- 15134406 TI - [Ergonomic analysis of work postures in the section of clothing and guidelines for planning of ergonomic solutions]. PMID- 15134408 TI - Crusaders or witch hunters. PMID- 15134409 TI - Combining technologies: a computerized occlusal analysis system synchronized with a computerized electromyography system. AB - Current advances in computer technologies have afforded dentists precision ways to examine occlusal contacts and muscle function. Recently, two separate computer technologies have been synchronized together, so that an operator can record their separate diagnostic data simultaneously. The two systems are: the T Scan II Occlusal Analysis System and the Biopak Electromyography Recording System. The simultaneous recording and playback capacity of these two computer systems allows the operator to analyze and correlate specific occlusal moments to specific electromyographic changes that result from these occlusal moments. This synchronization provides unparalleled evidence of the effect occlusal contact arrangement has on muscle function. Therefore, the occlusal condition of an inserted dental prosthesis or the occlusal scheme of the natural teeth (before and after corrective occlusal adjustments) can be readily evaluated, documented, and quantified for both, quality of occlusal parameters and muscle activity and the responses to the quality of the occlusal condition. This article describes their synchronization and illustrates their use in performing precision occlusal adjustment procedures on two patients: one who demonstrates occlusal disharmony while exhibiting the signs and symptoms of chronic myofascial pain dysfunction syndrome, and the other who had extensive restorative work accomplished but exhibits occlusal discomfort post-operatively. PMID- 15134410 TI - The longevity of temporomandibular disorder improvements after active treatment modalities. AB - Of concern to practitioners and patients alike are the long-term results following temporomandibular disorder/craniofacial pain (TMD/CP) treatments. This paper examines 270 patients who underwent active TMD/CP treatment and had follow up an average of 41 months after the termination of treatment when the patients had reached maximum medical improvement (MMI). Data show that, notwithstanding a mild degree of recrudescence, overall levels remain significantly lower than initial baseline symptoms, indicating a long-term benefit from active treatment. It could not be determined whether the recrudescent symptom levels were related to the conditions for which patients initially sought treatment or to new pathological states. Possible considerations for relapse may be the function of aging, musculoskeletal structural adaptation, resorting to parafunctional habits, and/or a change in psychosocial status. No significant differences were found between males and females regarding symptom levels and outcomes. Clinical experience suggests that TMD/CP symptoms do not resolve spontaneously and generally require active treatment. Unfortunately, most studies to date have largely measured symptom changes immediately (or very shortly) after the completion of treatment. This article, however, finds that the benefits of appropriate active TMD treatments remain long after treatment completion. PMID- 15134411 TI - Significance of frequency-selective fat saturation T2-weighted MR images for the detection of bone marrow edema in the mandibular condyle. AB - The objective of this study was to evaluate the utility of frequency-selective fat saturation (FS) T2-weighted images (T2WI) for the detection of bone marrow edema in the mandibular condyle. MR evidence of bone marrow abnormalities was examined on the set of FS T2WI and conventional T1WI or of conventional T2WI and T1WI in 200 patients with temporomandibular joint (TMJ) related pain. Other parameters studied were TMJ effusion, disk displacement categories, and cortical bone abnormalities. The detection rate and area of bone marrow edema by FS T2WI and T1WI were significantly greater than those assessed by conventional T2WI and T1WI. The correlation between bone marrow abnormalities on FS T2WI and T1WI and pain was significantly stronger than with conventional T2WI and T1WI. This study confirms that FS T2WI is useful for the detection of the "edema pattern" in the mandibular condylar associated with TMJ-related pain. PMID- 15134412 TI - Magnetic resonance evidence of joint effusion of the temporomandibular joint after fractures of the mandibular condyle: a preliminary report. AB - To investigate the clinical significance of magnetic resonance (MR) evidence of joint effusion of the temporomandibular joint after mandibular condylar fractures, magnetic resonance imaging (MRI) was performed on 18 joints in 15 patients with either unilateral or bilateral mandibular condylar fractures using a 1.5 Tesla MRI scanner (Signa, General Electric, Milwaukee, WI). MR evidence of joint effusion was evaluated and compared with the types and the positions of the fractures. MR evidence of joint effusion was observed in 11 of 18 TMJs, which was 61% of the condylar fractures. It appeared more frequently after fractures with dislocation than those without dislocation (p < 0.05). In addition, MR evidence of effusion appeared more frequently in TMJs after high condylar fractures (head to upper neck) than low condylar fractures (lower neck to subcondylar) (p < 0.05). These findings indicate that MR evidence of joint effusion may serve as a marker for the detection of severe intra-articular damage to the TMJ after mandibular condyle fractures. PMID- 15134413 TI - Role of the extracranial arteries in migraine headache: a review. AB - The pain of the migraine headache is often so debilitating that it severely compromises quality of life. The vascular component of the trigeminovascular system has been implicated in the pain mechanism. There is, however, debate as to whether the pain originates in the intracranial or extracranial vasculature or in both. In this article, evidence is presented to suggest that the extracranial arteries are the source of the pain in some migraine sufferers. PMID- 15134414 TI - Use of Theraflex-TMJ topical cream for the treatment of temporomandibular joint and muscle pain. AB - This randomized, double-blind study was designed to evaluate the effectiveness of the topical cream Theraflex-TMJ (NaBob/Rx, San Mateo, CA) in patients with masseter muscle pain and temporomandibular joint (TMJ) pain. Fifty-two subjects (5 males and 47 females) were instructed to apply a cream over the afflicted masseter muscle(s) or over the jaw joint(s) twice daily for two weeks. Theraflex TMJ cream was used by the experimental group, while a placebo cream was used by the control group. The means of pain ratings were calculated prior to the application of the cream (baseline), after ten days of tx (period 1), and 15 days of tx (period 2) days of treatment and five days after stopping the treatment (follow-up). There was a significant decrease in reported pain levels from baseline in the experimental group for period 1 (p < 0.01), period 2 (p < 0.001), and follow-up (p < 0.01). For the control group, no significant differences were found between the different time periods (p > 0.05). There was evidence of minor side effects such as skin irritation and/or burning on the site of the application in two subjects in the experimental as well as two subjects in the control groups. The data strongly suggest that Theraflex-TMJ topical cream is safe and effective for reducing pain in the masseter muscle and the temporomandibular joint. PMID- 15134415 TI - Electromyographic muscle EMG activity in mouth and nasal breathing children. AB - Mouth breathing may cause changes in muscle activity, because an upper airway obstruction leads may cause a person to extend his/her head forward, demanding a higher inspiratory effort on the accessory muscles (sternocleidomastoids). This purpose of this study is to compare, using electromyography (EMG), the activity pattern the sternocleidomastoid and upper trapezius muscles in mouth breathing children and nasal breathing children. Forty-six children, ages 8-12 years, 33 male and 13 female were included. The selected children were divided into two groups: Group I consisted of 26 mouth breathing children, and Group II, 20 nasal breathing children. EMG recordings were made using surface electrodes bilaterally in the areas of the sternocleidomastoideus and upper trapezius muscles, while relaxed and during maximal voluntary contraction. The data were analyzed using the Kruskall-Wallis statistical test. The results indicated higher activity during relaxation and lower activity during maximal voluntary contraction in mouth breathers when compared to the nasal breathers. It is suggested that the activity pattern of the sternocleidomastoid and upper trapezius muscles differs between mouth breathing children and nasal breathing children. This may be attributed to changes in body posture which causes muscular imbalance. Because of the limitations of surface EMG, the results need to be confirmed by adding force measurements and repeating the experiments with matched subjects. PMID- 15134416 TI - Improvement in smoothness of the chewing cycle following treatment of anterior crossbite malocclusion: a case report. AB - This is a report of a case in which improvement in masticatory jaw movement kinematics occurred following orthodontic treatment. A patient who demonstrated a skeletal Class II jaw relationship, with anterior crossbite between the right upper and lower lateral incisors, underwent treatment with an edgewise appliance. The trajectories and smoothness of the patient's jaw-closing movement were compared before and after orthodontic treatment. The correction of the anterior crossbite allowed the patient to consistently close the jaw with wider lateral excursion. Furthermore, after treatment, smoothness of the jaw-closing movements increased significantly, and the velocity profile was characterized as closer to that predicted by the minimum jerk (maximum smoothness) kinematic model. These findings suggest the value of trajectory smoothness (jerk-cost) as an objective indicator of kinematic improvement in gum chewing. In addition, the correction of anterior crossbite is demonstrated to enable the patient to perform smoother jaw closing movements during chewing. PMID- 15134418 TI - Mental health is fundamental to health. PMID- 15134417 TI - Chondroblastoma of the temporal bone involving the temporomandibular joint, mandibular condyle, and middle cranial fossa: case report and review of the literature. AB - Chondroblastoma is a highly destructive tumor, derived from immature cartilage cells, typically occurring in epiphyses of the long bones of adolescents and young adults. Those occurring in the temporal bone and TMJ area are likely to mimic TMJ symptoms. This report describes a unique case in which a chondroblastoma resulted in extensive destruction of the temporal bone, temporomandibular joint, mandibular condyle, and cranial base, including gross intracranial and extracranial involvement. With appropriate surgical management, the outcome for patients with chondroblastoma of the temporomandibular region is quite favorable. This case brings the total reported chondroblastomas to 59 in the temporal bone and eight in the mandibular condyle as of the date of submission of this article for publication. PMID- 15134419 TI - Making the most of your employer's retirement plan. PMID- 15134420 TI - Into the breach: a look at medical malpractice liability reform. PMID- 15134421 TI - Pathways to leadership conference: a professional development resource for women in medicine. PMID- 15134422 TI - When seniors face closed doors: are advance beneficiary notices the answer? PMID- 15134423 TI - Mandatory parental notification: the importance of confidential health care for adolescents. AB - Studies have shown that lack of confidentiality is a barrier to adolescents use of needed health care services. Professional medical organizations support confidential care for adolescents as a matter of individual and public health. Confidentiality is governed by both state and federal law, and physicians must be aware of both. Physicians must facilitate communication between teens and parents while guaranteeing confidential care to their adolescent patients. PMID- 15134426 TI - Incarcerated women and depression: a primer for the primary care provider. AB - Although women represent an increasing number of state prison inmates, they are studied less than their male counterparts are. Incarcerated women have higher rates of depression than both community samples and incarcerated men. The diagnosis and treatment of depression in incarcerated women is complicated by the presence of substance abuse, psychosocial stressors, medical problems, and personality disorders. This article focuses on the role of the community-based primary care provider in assessing, diagnosing, and treating the depressed female offender confined in state prisons. PMID- 15134425 TI - Depression in adolescent girls: screening and treatment strategies for primary care providers. AB - Adolescent girls are at a higher risk of depression than boys are. Depression is of particular concern in pregnant and postpartum adolescents because of the potential impact on the infant. Primary care providers (PCPs) should routinely screen adolescent girls for depression and consider depression in their differential diagnoses of somatic complaints. Both medications and psychotherapy are effective treatments for depression in adolescents. Interpersonal psychotherapy and cognitive therapy are both effective. PCPs have additional obligations to understand the possible barriers to adolescent use of mental health services in the community and to use available resources to advocate for appropriate health care for adolescents. PMID- 15134427 TI - Eating disorders: practical interventions. AB - The 4 currently recognized eating disorders are among the most common of psychiatric disorders to affect women in this country and are associated with chronic health impairments, although anorexia nervosa still carries a high mortality rate, including death by suicide. This review will focus primarily on the treatment of anorexia and bulimia nervosa and discuss interventions focused on engaging the patient, normalizing weight and nutritional status, changing dysfunctional attitudes and behaviors, and managing comorbid disorders such as major depression. PMID- 15134428 TI - Suicide among women: a critical review. AB - Despite increasing attention to suicide as a preventable outcome associated with mood disorders, little attention has been given to the risk factors for suicide among women. In this paper, we: 1) review the current literature regarding risk factors for suicide among women; 2) address the theories regarding risk and protective factors for women; 3) integrate the findings into a practical assessment of women's risk of suicide in clinical settings; and 4) consider avenues for future research. PMID- 15134429 TI - Menopause: neuroendocrine changes and hormone replacement therapy. AB - OBJECTIVE: We evaluated the effects of hormone replacement therapy (HRT) alone and in combination with a selective serotonin reuptake inhibitor on mood, cognition, and neuroendocrine parameters in peri- and postmenopausal women. METHODS: We measured neuroendocrine variations in peri- and postmenopausal depressed patients (DP) and postmenopausal normal control (NC) women (45 to 72 years old) before and after treatment with HRT alone and HRT combined with antidepressant medication. All subjects were without significant medical illness and off psychoactive or other medication that would interfere with neuroendocrine measures. RESULTS: Menopausal DP women reported greater severity of hot flashes, were less likely to be "morning" types, and had relatively good neuropsychological function compared with NC. DP and NC had comparable levels of reproductive hormones, with the exception of elevated prolactin levels, which increased, as did thyroid-stimulating hormone levels, in response to estradiol treatment. DP had poor sleep quality as measured both by subjective ratings and objective polysomnographic measures compared with NC. In DP estradiol did not enhance the effect of antidepressant alone on mood ratings. CONCLUSION: These findings may differ from other reports in the literature as a function of diagnoses of major depressive episode, randomized controlled trials, or dose and preparation of HRT. Further work is needed on the differential effect of treatment regimens in these disturbances that are evident primarily in baseline neuroendocrine function. PMID- 15134430 TI - Causing panic: anxieties surrounding obstetrical care. PMID- 15134424 TI - Women and bipolar disorder across the life span. AB - Bipolar I disorder occurs in approximately 1% of the adult population, and it affects women and men equally. Women develop bipolar II disorder, bipolar depression, mixed mania, and a rapid-cycling course of illness more commonly than men and are at greater risk of such comorbid conditions as alcohol use problems, thyroid disease, medication-induced obesity, and migraine headaches. The treatment of bipolar disorder remains challenging. Although lithium reduces symptoms and prevents recurrence with good efficacy, a significant number of patients stop taking it. Furthermore, several anticonvulsants and antidepressants are prescribed off label for acute episodes and prophylaxis despite the lack of adequate research support. Psychotherapy may alleviate mania or depression and improve treatment compliance, yet its ability to prevent relapse remains uncertain. Changes throughout the reproductive cycle also have an impact on the onset and presentation of bipolar symptoms and the choice of treatment. This article provides an overview of common presentations and comorbidities, along with approaches to evaluation and treatment of women with bipolar disorder. PMID- 15134431 TI - No need to be HAMLET or BAMLET to interact with histones: binding of monomeric alpha-lactalbumin to histones and basic poly-amino acids. AB - The ability of a specific complex of human alpha-lactalbumin with oleic acid (HAMLET) to induce cell death with selectivity for tumor and undifferentiated cells was shown recently to be mediated by interaction of HAMLET with histone proteins irreversibly disrupting chromatin structure [Duringer, C., et al. (2003) J. Biol. Chem. 278, 42131-42135]. Here we show that monomeric alpha-lactalbumin (alpha-LA) in the absence of fatty acids is also able to bind efficiently to the primary target of HAMLET, histone HIII, regardless of Ca(2+) content. Thus, the modification of alpha-LA by oleic acid is not required for binding to histones. We suggest that interaction of negatively charged alpha-LA with the basic histone stabilizes apo-alpha-LA and destabilizes the Ca(2+)-bound protein due to compensation for excess negative charge of alpha-LA's Ca(2+)-binding loop by positively charged residues of the histone. Spectrofluorimetric curves of titration of alpha-LA by histone H3 were well approximated by a scheme of cooperative binding of four alpha-LA molecules per molecule of histone, with an equilibrium dissociation constant of 1.0 microM. Such a stoichiometry of binding implies that the binding process is not site-specific with respect to histone and likely is driven by just electrostatic interactions. Co-incubation of positively charged poly-amino acids (poly-Lys and poly-Arg) with alpha-LA resulted in effects which were similar to those caused by histone HIII, confirming the electrostatic nature of the alpha-LA-histone interaction. In all cases that were studied, the binding was accompanied by aggregation. The data indicate that alpha lactalbumin can be used as a basis for the design of antitumor agents, acting through disorganization of chromatin structure due to interaction between alpha LA and histone proteins. PMID- 15134432 TI - The C-terminal domain of the betaine carrier BetP of Corynebacterium glutamicum is directly involved in sensing K+ as an osmotic stimulus. AB - The glycine betaine carrier BetP of Corynebacterium glutamicum was recently shown to function both as an osmosensor and as an osmoregulator in proteoliposomes by sensing changes in the internal K(+) concentration as a measure of hyperosmotic stress. In vivo analysis of mutants carrying deletions at the C-terminal extension of BetP indicated that this domain participates in osmostress-dependent activity regulation. To address the question, whether a putative K(+) sensor is located within the C-terminal domain, several mutants with truncations in this domain were purified and reconstituted in proteoliposomes of Escherichia coli phospholipids, since this in vitro system allowed variation of the K(+) concentration at the lumenal side. Truncation of 12 amino acids led to a partly deregulated BetP in terms of osmoregulation; however, K(+) sensitivity was not impaired in this mutant. The deletion of 25 amino acid residues at the C-terminal end of BetP led to both deregulation of the carrier activity, i.e., high activity independent of external osmolality, and loss of K(+)-dependent transport stimulation, indicating that this region of the C-terminal domain is necessary for K(+) sensing and/or K(+)-dependent carrier activation. Immunological and proteolysis analyses showed that BetP and its recombinant forms were reconstituted in a right-side-out orientation, i.e., the C-terminal domain faces the lumen of the proteoliposomes and is thus able to detect the K(+) signal at the inside. This is the first experimental demonstration of a direct connection between an osmotic stimulus, i.e., the change in internal K(+), and a putative sensor domain. PMID- 15134433 TI - Nucleotide excision repair defect influences lethality and mutagenicity induced by Me-lex, a sequence-selective N3-adenine methylating agent in the absence of base excision repair. AB - Using a yeast shuttle vector system, we have previously reported on the toxicity and mutagenicity of Me-lex, [1-methyl-4-[1-methyl-4-[3 (methoxysulfonyl)propanamido]pyrrole-2-carboxamido]pyrrole-2-carboxamido]propane, a compound that selectively generates 3-methyladenine (3-MeA). We observed that a mutant strain defective in Mag1, the glycosylase that excises 3-MeA in the initial step of base excision repair (BER) to generate an abasic site, is significantly more sensitive to the toxicity of Me-lex with respect to wild type but shows only a marginal increase in mutagenicity. A strain defective in AP endonuclease activity (Deltaapn1apn2), also required for functional BER, is equally sensitive to the toxicity as the Deltamag1 mutant but showed a significantly higher mutation frequency. In the present work, we have explored the role of nucleotide excision repair (NER) in Me-lex-induced toxicity and mutagenicity since it is known that NER acts on abasic sites in vivo in yeast and in vitro assays. To accomplish this, we have deleted one of the genes essential for NER in yeast, namely, RAD14, both in the context of an otherwise DNA repair proficient strain (Deltarad14) and in a BER-defective isogenic derivative lacking the MAG1 gene (Deltamag1rad14). Interestingly, no sensitivity to the treatment with Me-lex was conferred by the simple deletion of RAD14. However, a significant enhancement in toxicity and mutagenicity was observed when cells lacked both Rad14 and Mag1. The mutation spectrum induced by Me-lex in the Deltamag1rad14 strain is indistinguishable from that observed in the Deltaapn1/Deltaapn2 or in the Deltamag1 strains. The results indicate that in yeast NER can play a protective role against 3-MeA-mediated toxicity and mutagenicity; however, the role of NER is appreciable only in a BER-defective background. PMID- 15134435 TI - How C-terminal carboxyamidation alters the biological activity of peptides from the venom of the eumenine solitary wasp. AB - Inflammatory peptides display different types of post-transcriptional modifications, such as C-terminal amidation, that alter their biological activity. Here we describe the structural and molecular dynamics features of the mast cell degranulating peptide, eumenine mastoparan-AF (EMP-AF-NH(2)), found in the venom of the solitary wasp, and of its carboxyl-free C-terminal form (EMP-AF COO(-)) characterized by a reduced activity. Circular dichroism indicates that both peptides switch from a random coil conformation in water to a helical structure in TFE and SDS micelles. NMR data, in 30% TFE, reveal that the two peptides fold into an alpha-helix spanning most of their length, while they differ in terms of molecular rigidity. To understand the origins of the conformational flexibility observed in the case of EMP-AF-COO(-), a 5 ns MD simulation was carried out for each peptide, in an explicit water/TFE environment. The results show that the two peptides differ in an H-bond between Leu14 NH(2) and the backbone carbonyl of Ile11. The loss of that H-bond in EMP-AF COO(-) leads to a significant modification of its structural dynamics. In fact, as evidenced by essential dynamics analysis, while EMP-AF-NH(2) exists mainly as a rigid structure, EMP-AF-COO(-) presents two helical stretches that fluctuate in some sort of independent fashion. We conclude that the diverse biological activity of the two peptides is not simply due to the reduction of the net positive charge, as generally suggested, but also to a structural perturbation of the amphipathic alpha-helix that affects their ability to perturb the cell membrane. PMID- 15134434 TI - Role of the Rab11-associated intracellular pool of receptors formed by constitutive endocytosis of the beta isoform of the thromboxane A2 receptor (TP beta). AB - Intracellular trafficking pathways of G protein-coupled receptors (GPCRs), following their agonist-induced endocytosis and their consequences on receptor function, are the subject of intense research efforts. However, less is known regarding their constitutive endocytosis. We previously demonstrated that the beta isoform of the thromboxane A(2) receptor (TPbeta) undergoes constitutive and agonist-induced endocytosis. Constitutive endocytosis of GPCRs can lead to the formation of an intracellular pool of receptors from which they can recycle back to the cell surface. In the present report, we show with the help of two TPbeta mutants (TPbeta-Y339A and TPbeta-I343A) specifically deficient in constitutive endocytosis that this intracellular pool of receptors serves to maintain agonist sensitivity over prolonged receptor stimulation in HEK293 cells. Second messenger generation by the TPbeta-Y339A and TPbeta-I343A mutants was drastically reduced compared to the wild-type receptor as suggested by dose-response and time-course experiments of inositol phosphates production following agonist treatment, despite normal coupling between the receptors and the Galpha(q) protein. Moreover, second messenger production after receptor activation was dramatically reduced when cells were pretreated with monensin, a recycling inhibitor. Receptor cell surface expression and endocytosis experiments further revealed that the small GTPase Rab11 protein is a determinant factor in controlling TPbeta recycling back to the cell surface. Co-localization experiments performed by immunofluorescence microscopy indicated that both constitutive and agonist triggered endocytosis resulted in targeting of TPbeta to the Rab11-positive recycling endosome. Thus, we provide evidence that constitutive endocytosis of TPbeta forms a pool of receptors in the perinuclear recycling endosome from which they recycle to the cell surface, a process involved in preserving receptor sensitivity to agonist stimulation. PMID- 15134436 TI - Transferrin-modified liposomes equipped with a pH-sensitive fusogenic peptide: an artificial viral-like delivery system. AB - Liposomes are one of the most promising systems for selective cellular targeting via introduction of specific ligands for cell-surface receptors. After being taken up by the cells, these liposomes usually follow intracellular pathways of receptor-mediated endocytosis. Control of intracellular trafficking is required for optimized drug delivery. In this study, we elucidated the intracellular fate of transferrin-modified liposomes and succeeded in altering it by introducing the pH-sensitive fusogenic peptide, GALA (WEAALAEALAEALAEHLAEALAEALEALAA). Transferrins that are chemically attached to a liposomal surface (Tf-L) were internalized via receptor-mediated endocytosis more slowly than unmodified transferrins. In contrast to the recyclable nature of transferrin, liposome attached transferrins together with encapsulated rhodamines were retained in vesicular compartments. When GALA was introduced into liposomal membranes using a cholesteryl moiety for anchoring (Chol-GALA), rhodamines were efficiently released and diffused into the cytosol. The addition of GALA to the Tf-L containing medium or the encapsulation of GALA in Tf-L did not induce similar effects. These results clearly indicate that GALA must be present on the surface of liposomes to exert its function. In vitro energy transfer and dynamic light scattering experiments suggested that the endosomal escape of the encapsulates in Tf-L equipped with Chol-GALA can be attributed to pH-dependent membrane fusion. With GALA present on the surface, intracellular trafficking of liposomes after receptor-mediated endocytosis could be successfully controlled. PMID- 15134437 TI - Differential effects of substrate on type I and type II PKA holoenzyme dissociation. AB - It has been widely accepted that cAMP activates the protein kinase A (PKA) holoenzyme by dissociating the regulatory and catalytic subunits, thus freeing the catalytic subunit to phosphorylate its targets. However, recent experiments suggest that cAMP does not fully dissociate the holoenzyme. Here, we investigate this mechanism further by using small-angle X-ray scattering to study, at physiological enzyme concentrations, the type Ialpha and type IIbeta holoenzyme structures under equilibrium solution conditions without any labeling of the protein subunits. We observe that while the addition of a molar excess of cAMP to the type Ialpha PKA holoenzyme causes partial dissociation, it is only upon addition of a PKA peptide substrate together with cAMP that full dissociation occurs. Similarly, addition of excess cAMP to the type IIbeta holoenzyme causes only a partial dissociation. However, while the addition of peptide substrate as well as excess cAMP causes somewhat more dissociation, a significant percentage of intact type IIbeta holoenzyme remains. These results confirm that both the type Ialpha and the type IIbeta holoenzymes are more stable in the presence of cAMP than previously thought. They also demonstrate that substrate plays a differential role in the activation of type I versus type II holoenzymes, which could explain some important functional differences between PKA isoforms. On the basis of these data and other recently published data, we propose a structural model of type I holoenzyme activation by cAMP. PMID- 15134438 TI - Interaction of the retinal insulin receptor beta-subunit with the p85 subunit of phosphoinositide 3-kinase. AB - Recently, we have shown that phosphoinositide 3-kinase (PI3K) in retina is regulated in vivo through light activation of the insulin receptor beta-subunit. In this study, we have cloned the 41 kDa cytoplasmic region of the retinal insulin receptor (IRbeta) and used the two-hybrid assay of protein-protein interaction in the yeast Saccharomyces cerevisiae to demonstrate the interaction between the p85 subunit of PI3K and the cytoplasmic region of IRbeta. Under conditions where IRbeta autophosphorylates, substitution of Y1322F and M1325P in IRbeta resulted in the abolition of p85 binding to the IRbeta, confirming that the p85 subunit of PI3K binds to Y1322. The binding site for p85 on IRbeta was also confirmed in the yeast three-hybrid system. Using the C-terminal region of IRbeta (amino acids 1293-1343 encompassing the YHTM motif) as bait and supplying an exogenous tyrosine kinase gene to yeast cells, we determined that the IRbeta pYTHM motif interacts with p85. We also used retinal organ cultures to demonstrate insulin activation of the insulin receptor and subsequent binding of p85, measured through GST pull-down assays with p85 fusion proteins. Further, the Y960F mutant insulin receptor, which does not bind IRS-1, is capable of bringing down PI3K activity from retina lysates. On the other hand, in response to insulin, IRS-2 is able to interact with the p85 subunit of PI3K in the retina. These results suggest that multiple signaling pathways could regulate the PI3K activity and subsequent activation of Akt in the retina. PMID- 15134439 TI - Role of the isoprenyl pocket of the G protein beta gamma subunit complex in the binding of phosducin and phosducin-like protein. AB - Phosducin (Pdc) and phosducin-like protein (PhLP) regulate G protein-mediated signaling by binding to the betagamma subunit complex of heterotrimeric G proteins (Gbetagamma) and removing the dimer from cell membranes. The binding of Pdc induces a conformational change in the beta-propeller structure of Gbetagamma, creating a pocket between blades 6 and 7. It has been proposed that the isoprenyl group of Gbetagamma inserts into this pocket, stabilizing the Pdc.Gbetagamma structure and decreasing the affinity of the complex for the lipid bilayer. To test this hypothesis, the binding of Pdc and PhLP to several Gbetagamma dimers containing variants of the beta or gamma subunit was measured. These variants included modifications of the isoprenyl group (gamma), residues involved in the conformational change (beta), and residues lining the proposed prenyl pocket (beta). Switching prenyl groups from farnesyl to geranylgeranyl or vice versa had little effect on binding. However, alanine substitution of one residue in the beta subunit involved in the conformational change (W332) decreased binding 5-fold. Alanine substitution of certain residues within the prenyl pocket caused only minor decreases in binding, while a lysine substitution of T329 within the pocket inhibited binding 10-fold. Molecular modeling of the binding energy of the Pdc.Gbeta(1)gamma(2) complex required insertion of the geranylgeranyl group into the prenyl pocket in order to accurately predict the effects of prenyl pocket amino acid substitutions. Finally, a dimer containing a gamma subunit with no prenyl group (gamma(2)-C68S) decreased binding by nearly 20 fold. These results support the structural model in which the prenyl group escapes contact with the aqueous milieu by inserting into the prenyl pocket and stabilizing the Pdc-binding conformation of Gbetagamma. PMID- 15134440 TI - Inhibition of protein interactions with the beta 2 sliding clamp of Escherichia coli DNA polymerase III by peptides from beta 2-binding proteins. AB - The sliding clamp of the Escherichia coli replisome is now understood to interact with many proteins involved in DNA synthesis and repair. A universal interaction motif is proposed to be one mechanism by which those proteins bind the E. coli sliding clamp, a homodimer of the beta subunit, at a single site on the dimer. The numerous beta(2)-binding proteins have various versions of the consensus interaction motif, including a related hexameric sequence. To determine if the variants of the motif could contribute to the competition of the beta-binding proteins for the beta(2) site, synthetic peptides derived from the putative beta(2)-binding motifs were assessed for their abilities to inhibit protein beta(2) interactions, to bind directly to beta(2), and to inhibit DNA synthesis in vitro. A hierarchy emerged, which was consistent with sequence similarity to the pentameric consensus motif, QL(S/D)LF, and peptides containing proposed hexameric motifs were shown to have activities comparable to those containing the consensus sequence. The hierarchy of peptide binding may be indicative of a competitive hierarchy for the binding of proteins to beta(2) in various stages or circumstances of DNA replication and repair. PMID- 15134441 TI - Effect of phosphorothioate chirality on i-motif structure and stability. AB - The P-chiral stereo-defined phosphorothioate groups have been introduced into all of the four internucleotide positions of d(T(PS1)C(PS2)C(PS3)C(PS4)C) (PSn = phosphorothioate group), and among the 16 possible diastereomers of PS-d(TC(4)), 10 stereomers have been synthesized to investigate the effects of the sense of the P-chirality upon the structure and stability of the i-motif structure. The temperature dependence of circular dichroism spectra showed that the melting temperature (T(m)) of the [all R(p)]-PS-d(TC(4)) i-motifs was 31 degrees C, identical to that of the parent oligomer, PO-d(TC(4)), while that of the [all S(p)]-PS-d(TC(4)) i-motif was largely decreased by 11 degrees C. Single substitution of R(p) with S(p) caused a decrease of T(m) by 3-4 degrees C at positions of PS1, PS2, and PS3 and by 1 degrees C at that of PS4, showing the additive property of the T(m) suppression. The comparison of the NOESY spectra between [all R(p)]-PS-, [all S(p)]-PS-, and PO-d(TC(4)) showed that intraresidual H6-H3' and H2' '-H4' NOE cross-peaks of the all S(p) isomer are weaker than those of the all R(p) isomer and PO-d(TC(4)), indicating the change in the C3'-endo conformation and glycosidic bond angle. The structural alternation for the i motif formed by [all S(p)]-PS-d(TC(4)) is also suggested by the chemical shift differences of C2/C3/C4 H2''and H4' protons from those of [all R(p)]-PS-d(TC(4)) and PO-d(TC(4)). These results suggest that the S(p) configuration at phosphorus of the phosphorothioate linkage changes the sugar-phosphorothioate conformation and intermolecular interaction in the narrow groove, leading to the destabilization of the i-motif structure. PMID- 15134442 TI - Mechanism of RNA 2'-O-methylation: evidence that the catalytic lysine acts to steer rather than deprotonate the target nucleophile. AB - The weight of current evidence suggests that RNA 2'-O-MTases employ an S(N)2 mechanism with an in-line attack of the target nucleophile upon the methyl group of the AdoMet cofactor. It has been suggested that, like the phosphohydrolytic enzymes, ribozymes, and nucleic acid polymerases, the RNA 2'-O-MTases initially activate the substrate's attacking hydroxyl oxygen by deprotonation. Here, evidence is presented that the vaccinia virus mRNA cap specific 2'-O-MTase VP39 does not promote RNA 2'-oxyanion formation but that instead it acts by steering a hydroxyl oxygen orbital toward the cofactor methyl center. PMID- 15134443 TI - 5-Formyluracil-induced perturbations of DNA function. AB - Oxidation of the thymine methyl group can generate 5-formyluracil (FoU), which is known to be both mutagenic and chemically unstable in DNA. Synthetic oligonucleotides containing FoU at defined sites have been prepared to investigate potential mechanisms by which FoU might perturb DNA function. The half-life of the glycosidic bond of an FoU residue in single-stranded DNA under physiological conditions of temperature and pH is estimated to be approximately 148 days, orders of magnitude shorter than the parent pyrimidine, thymine. This reduced stability of FoU residues in DNA is attributed to the inductive properties of the 5-formyl substituent. Oxidative modification of the thymine methyl group could also inhibit association with sequence-specific DNA-binding proteins. Alternatively, the 5-formyl substituent of FoU could cross-link nonspecifically with protein amino groups. Transcription factor AP-1 is known to make specific contacts with thymine methyl groups of DNA in its recognition sequence. Substitution of T by FoU is shown to inhibit AP-1 (c-Jun homodimer) binding with a DeltaDeltaG of approximately 0.6 kcal/mol. No evidence of cross link formation is observed with either AP-1 or polylysine. Molecular modeling studies on the FoU-containing oligonucleotide sequence corresponding to the duplex used in the experimental studies demonstrate that the 5-formyl substituent of an FoU residue paired with adenine lies in the plane of the pyrimidine base and is well protected from solvent on one face and only partially accessible on the other. The results of this study suggest that although FoU residues in DNA are considerably more labile than thymine, they are likely to be present long enough to miscode as well as interfere with DNA-protein interactions. PMID- 15134444 TI - Semisynthesis of phosphovariants of Smad2 reveals a substrate preference of the activated T beta RI kinase. AB - Transforming growth factor-beta (TGF-beta) signaling regulates a wide range of cellular processes. Aberrant TGF-beta signaling has been implicated in various disease states in humans. A key element in this signaling pathway is phosphorylation of R-Smads such as Smad2 at the last two serine residues of the C terminal sequence CSSXS (residues 463-467 in Smad2) by the TbetaRI receptor kinase. Phosphorylation results in the release of the R-Smad from the membrane anchored protein SARA, binding to the co-mediator protein Smad4, translocation into the nucleus, and regulation of target gene expression. Expressed protein ligation was used to probe the contribution of the individual phosphate groups to Smad2 oligomerization and phosphorylation by TbetaRI. Phosphorylation at both positions was required to generate a stable homotrimer; however, the driving force for Smad2 self-association is provided by pSer465. Additionally, SARA was found to modulate the self-association of partially phosphorylated Smad2, which suggests an added role for this protein in preventing premature release of a monophosphorylated substrate from the receptor complex. In related studies, prephosphorylation of Smad2 at Ser465 was found to significantly increase the rate of phosphorylation at Ser467, suggesting that there may be specific recognition determinants within the kinase for the monophosphorylated intermediate. This information was exploited to design an improved peptide substrate for TbetaRI, which may prove valuable in the design of inhibitors of the TGF-beta pathway. PMID- 15134445 TI - Mechanism for the hydrolysis of organophosphates by the bacterial phosphotriesterase. AB - Phosphotriesterase (PTE) from Pseudomonas diminuta is a zinc metalloenzyme that hydrolyzes a variety of organophosphorus compounds. The kinetic parameters of Zn/Zn PTE, Cd/Cd PTE, and a mixed-metal Zn/Cd hybrid PTE were obtained with a variety of substrates to determine the role of each metal ion in binding and catalysis. pH-rate profiles for the hydrolysis of diethyl p-nitrophenyl phosphate (I) and diethyl p-chlorophenyl phosphate (II) demonstrated that the ionization of a single group in the pH range of 5-10 was critical for substrate turnover. The pK(a) values determined from the kinetic assays were dependent on the identity of the metal ion that occupied the alpha site within the binuclear metal center. These results suggest that the hydrolytic nucleophile is activated as a hydroxide via the ionization of a water molecule attached to the alpha-metal ion. The kinetic constants for the hydrolysis of II and diethyl p-chlorophenyl thiophosphate (IV) were determined for the metal substituted forms of PTE. The kinetic constants for IV were greater than those for II. The inverse thio effect is consistent with the polarization of the phosphoryl oxygen/sulfur bond via a direct ligation to the metal center. The rate enhancement is greater when Cd(2+) occupies the beta-metal-ion position. A series of alanine and asparagine mutations were used to characterize the catalytic roles of Asp233, His254, and Asp301. Mutations to either Asp233 or His254 resulted in an enhanced rate of hydrolysis for the sluggish substrate, diethyl p-chlorophenyl phosphate, and a decrease in the kinetic constants for paraoxon (I). These results are consistent with the existence of a proton relay from Asp301 to His254 to Asp233 that is used to ferry protons away from the active site with substrates that do not require activation of the leaving group phenol. A mechanism for the hydrolysis of organophosphates by the bacterial PTE has been proposed. PMID- 15134446 TI - Evolution of enzymatic activity in the enolase superfamily: structural studies of the promiscuous o-succinylbenzoate synthase from Amycolatopsis. AB - Divergent evolution of enzyme function is commonly explained by a gene duplication event followed by mutational changes that allow the protein encoded by the copy to acquire a new function. An alternate hypothesis is that this process is facilitated when the progenitor enzyme acquires a second function while maintaining the original activity. This phenomenon has been suggested to occur in the o-succinylbenzoate synthase (OSBS) from a species of Amycolatopsis that catalyzes not only the physiological syn-dehydration reaction of 2-succinyl 6-hydroxy-2,4-cyclohexadiene-1-carboxylate but also an accidental racemization of N-acylamino acids [Palmer, D. R., Garrett, J. B., Sharma, V., Meganathan, R., Babbitt, P. C., and Gerlt, J. A. (1999) Biochemistry 38, 4252-4258]. To understand the molecular basis of this promiscuity, three-dimensional structures of liganded complexes of this enzyme have been determined, including the product of the OSBS reaction and three N-acylamino acid substrates for the N-acylamino acid racemase (NAAAR) reaction, N-acetylmethionine, N-succinylmethionine, and N succinylphenylglycine, to 2.2, 2.3, 2.1, and 1.9 A resolution, respectively. These structures show how the active-site cavity can accommodate both the hydrophobic substrate for the OSBS reaction and the substrates for the accidental NAAAR reaction. As expected, the N-acylamino acid is sandwiched between lysines 163 and 263, which function as the catalytic bases for the abstraction of the alpha-proton in the (R)- and (S)-racemization reactions, respectively [Taylor Ringia, E. A., Garrett, J. B, Thoden, J. B., Holden, H. M., Rayment, I., and Gerlt, J. A. (2004) Biochemistry 42, 224-229]. Importantly, the protein forms specific favorable interactions with the hydrophobic amino acid side chain, alpha carbon, carboxylate, and the polar components of the N-acyl linkage. Accommodation of the components of the N-acyl linkage appears to be the reason that this enzyme is capable of a racemization reaction on these substrates, whereas the orthologous OSBS from Escherichia coli lacks this functionality. PMID- 15134448 TI - Oxygen activation by the noncoupled binuclear copper site in peptidylglycine alpha-hydroxylating monooxygenase. Spectroscopic definition of the resting sites and the putative CuIIM-OOH intermediate. AB - Spectroscopic methods, density functional calculations, and ligand field analyses are combined to define the geometric models and electronic structure descriptions of the Cu(M) and Cu(H) sites in the oxidized form of the noncoupled binuclear copper protein peptidylglycine alpha-hydroxylating monooxygenase (PHM). The Cu(M) site has a square pyramidal geometry with a long axial Cu-methionine bond and two histidines, H(2)O, and OH(-) as equatorial ligands. The Cu(H) site has a slightly D(2)(d) distorted square planar geometry with three histidines and H(2)O ligands. The structurally inequivalent Cu(M) and Cu(H) sites do not exhibit measurable differences in optical and electron paramagnetic resonance (EPR) spectra, which result from their similar ligand field transition energies and ground-state Cu covalencies. The additional axial methionine ligand interaction and associated square pyramidal distortion of the Cu(M) site have the opposite effect of the strong equatorial OH(-) donor ligand on the Cu d orbital splitting pattern relative to the Cu(H) site leading to similar ligand field transition energies for both sites. The small molecule NO(2)(-) binds in different coordination modes to the Cu(M) and Cu(H) site because of differences in their exchangeable coordination positions resulting in these Cu(II) sites being spectroscopically distinguishable. Azide binding to PHM is used as a spectroscopic and electronic structure analogue to OOH(-) binding to provide a starting point for developing a geometric and electronic structural model for the putative Cu(II)(M)-OOH intermediate in the H-atom abstraction reaction of PHM. Possible electronic structure contributions of the Cu(II)(M)-OOH intermediate to reactivity are considered by correlation to the well-studied L3Cu(II)-OOH model complex (L3 = [HB[3-tBu-5-iPrpz](3)]). The Met-S ligand of the Cu(M) site is found to contribute to the stabilization of the Cu(II)(M)-oxyl species, which would be a product of Cu(II)(M)-OOH H-atom abstraction reaction. This Met-S contribution could have a significant effect on the energetics of a H-atom abstraction reaction by the Cu(II)(M)-OOH intermediate. PMID- 15134447 TI - Evidence for a proton transfer network and a required persulfide-bond-forming cysteine residue in Ni-containing carbon monoxide dehydrogenases. AB - Carbon monoxide dehydrogenase from Moorella thermoacetica catalyzes the reversible oxidation of CO to CO(2) at a nickel-iron-sulfur active site called the C-cluster. Mutants of a proposed proton transfer pathway and of a cysteine residue recently found to form a persulfide bond with the C-cluster were characterized. Four semiconserved histidine residues were individually mutated to alanine. His116 and His122 were essential to catalysis, while His113 and His119 attenuated catalysis but were not essential. Significant activity was "rescued" by a double mutant where His116 was replaced by Ala and His was also introduced at position 115. The activity was also rescued in double mutants where His122 was replaced by Ala and His was simultaneously introduced at either position 121 or position 123. Activity was also rescued by replacing His with Cys at position 116. Mutation of conserved Lys587 near the C-cluster attenuated activity but did not eliminate it. Activity was virtually abolished in a double mutant where Lys587 and His113 were both changed to Ala. Mutations of conserved Asn284 also attenuated activity. These effects suggest the presence of a network of amino acid residues responsible for proton transfer rather than a single linear pathway. The Ser mutant of the persulfide-forming Cys316 was essentially inactive and displayed no electron paramagnetic resonance signals originating from the C cluster. Electronic absorption and metal analysis suggest that the C-cluster is absent in this mutant. The persulfide bond appears to be essential for either the assembly or the stability of the C-cluster, and possibly for eliciting the redox chemistry of the C-cluster required for catalytic activity. PMID- 15134449 TI - Role of the conserved arginine pair in proton and electron transfer in cytochrome C oxidase. AB - A hydrogen-bonded network is observed above the hemes in all of the high resolution crystal structures of cytochrome oxidases. It includes water and a pair of arginines, R481 and R482 (Rhodobacter sphaeroides numbering), that interact directly with heme a and the heme a(3) propionates. The hydrogen-bonded network provides potential pathways for proton release. The arginines, and the backbone peptide bond between them, have also been proposed to form part of a facilitated electron transfer route between Cu(A) and heme a. Our studies show that mutations of R482 (K, Q, and A) and R481 (K) retain substantial activity and are able to pump protons, but at somewhat reduced rates and stoichiometries. A slowed rate of electron transfer from cytochrome c to Cu(A) suggests a change in the orientation of cytochrome c binding in all but the R to K mutants. The mutant R482P is more perturbed in its structure and is altered in the redox potential difference between heme a and Cu(A): +18 mV for R482P and +46 mV for the wild type (heme a - Cu(A)). The electron transfer rate between Cu(A) and heme a is also altered from 93000 s(-1) in the wild type to 50 s(-1) in the oxidized R482P mutant, reminiscent of changes observed in a Cu(A)-ligand mutant, H260N. In neither case is the approximately 2000-fold change in the rate accounted for by the altered redox potentials, suggesting that both cause a major modification in the path or reorganization energy of electron transfer. PMID- 15134450 TI - NMR structures of loop B RNAs from the stem-loop IV domain of the enterovirus internal ribosome entry site: a single C to U substitution drastically changes the shape and flexibility of RNA. AB - The 5'-untranslated region of positive-strand RNA viruses harbors many cis-acting RNA structural elements that are important for various viral processes such as replication, translation, and packaging of new virions. Among these is loop B RNA of the stem-loop IV domain within the internal ribosomal entry site (IRES) of enteroviruses, including Poliovirus type 1 (PV1). Studies on PV1 have shown that specific recognition of loop B by the first KH (hnRNP K homology) domain of cellular poly(rC)-binding protein 2 (PCBP2) is essential for efficient translation of the viral mRNA. Here we report the NMR solution structures of two representative sequence variants of enteroviral loop B RNA. The two RNA variants differ at only one position (C vs U) within a six-nucleotide asymmetric internal loop sequence that is the binding site for the PCBP2 KH1 domain. Surprisingly, the two RNAs are drastically different in the overall shape and local dynamics of the bulge region. The RNA with the 5'-AUCCCU bulge sequence adopts an overall L shape. Its bulge nucleotides, especially the last four, are highly flexible and not very well defined by NMR. The RNA with the 5'-AUUCCU bulge sequence adopts an overall U shape, and its bulge sequence exhibits only limited flexibility. A detailed analysis of the two RNA structures and their dynamic properties, as well as available sequence data and known KH domain-RNA complex structures, not only provides insights into how loop B RNA might be recognized by the PCBP2 KH1 domain but also suggests a possible correlation between structural flexibility and pre existing structural features for protein recognition. PMID- 15134451 TI - NMR and mutagenesis studies on the phosphorylation region of human cardiac troponin I. AB - Phosphorylation of the cardiac troponin complex by PKA at S22 and S23 of troponin I (TnI) accelerates Ca(2+) release from troponin C (TnC). The region of TnI around the bisphosphorylation site binds to, and stabilizes, the Ca(2+) bound N terminal domain of TnC. Phosphorylation interferes with this interaction between TnI and TnC resulting in weaker Ca(2+) binding. In this study, we used (1)H-(15)N HSQC NMR to investigate at the atomic level the interaction between an N-terminal fragment of TnI consisting of residues 1-64 of TnI (I1-64) and TnC. We produced several mutants of I1-64, TnI, and TnC to test the contribution of certain residues to the transmission of the phosphorylation signal in both NMR experiments and functional assays. We also investigated how phosphorylation of the PKC sites in I1-64 (S41 and S43) affects the interaction of I1-64 with TnC. We found that phosphorylation of S22 and S23 produced only localized effects in the structure of I1-64 between residues 24 and 34. Residues 1-17 of I1-64 did not bind to TnC, and residues 38-64 bound tightly to the C-terminal domain of TnC regardless of phosphorylation. Residues 22-34 bound weakly to TnC in a phosphorylation sensitive manner. Bisphosphorylation prevented this phosphorylation switch region from interacting with TnC. Systematic mutation of residues in the phosphorylation switch did not prevent PKA phosphorylation from accelerating Ca(2+) release from troponin. We conclude that the phosphorylation switch binds to TnC via an extended interaction site spanning residues R19 to A34. PMID- 15134452 TI - Interfacial folding and membrane insertion of a designed helical peptide. AB - Nonconstitutive membrane-active proteins, such as diphtheria toxin, must refold on membrane interfaces in the course of membrane penetration. A useful step in deciphering this process is to understand quantitatively the energetics of interface-mediated insertion of model transmembrane helices. A difficulty is that peptides that are sufficiently hydrophobic to span a lipid bilayer have a strong tendency to aggregate in the aqueous phase. To learn how to control the aqueous and membrane behavior of model peptides, we designed a 31-residue peptide (TMX-3) whose properties are described here. TMX-3 has two important structural features: a proline residue in the hydrophobic core that discourages the formation of highly helical aggregates in solution and two histidine residues that allow control of membrane and solution interactions by means of pH changes. The partitioning of TMX-3 into membranes followed complex kinetics, induced helicity, and shifted the histidine pK(a) from 6.8 to approximately 6. Topology measurements disclosed two general modes of TMX-3 binding: interfacial (IF) at low peptide concentrations and partial transmembrane (TM) insertion at higher concentrations. Both modes were reversible and, consequently, suitable for thermodynamic analysis. The free energies of IF partitioning of TMX-3 with deprotonated (pH 7.6) and protonated histidines (pH 4.5) were estimated by fluorescence titration to be -6.7 and -5.0 kcal/mol, respectively. These results show that histidine titration is likely to be important in the pH-dependent refolding of toxins on membrane interfaces and that the most favored state of TMX 3 under any conditions is the IF folded state, which emphasizes the importance of such states in the spontaneous refolding and insertion of diphtheria and other membrane toxins. PMID- 15134453 TI - Comparison between catalase-peroxidase and cytochrome c peroxidase. The role of the hydrogen-bond networks for protein stability and catalysis. AB - A detailed resonance Raman and electronic absorption investigation has been carried out on a series of novel distal and proximal variants of recombinant catalase-peroxidase from the cyanobacterium Synechocystis PCC 6803. In particular, variants of the distal triad Pro-Asp-Asn and the proximal triad His Asp-Trp have been studied in their ferric and ferrous states at various pH. The data suggest marked differences in the structural role of the conserved residues and hydrogen-bond networks in KatG and CCP, which might be connected to the different catalytic activity. In particular, in KatG the proximal residues have a major role in the stability of the protein architecture because the disruption of the proximal Trp-Asp hydrogen bond by mutation weakens heme binding to the protein. On the distal side, replacing the hydrogen-acceptor carboxamide group of Asn153 by an aspartate carboxylate group or an aliphatic residue alters or disrupts the hydrogen bond with the distal His. As a consequence, the basicity of His123 is altered. The effect of mutation on Asp152 is noteworthy. Replacement of the Asp152 with Ser makes the architecture of the protein very similar to that of CCP. The Asp152 residue, which has been shown to be important in the hydrogen peroxide oxidation reaction, is expected to be hydrogen bonded to the nitrogen atom of Ile248 which is part of the KatG-specific insertion LL1, as in other KatGs. This insertion is at one edge of the heme, and connects the distal side with the proximal helices E and F, the latter carrying the proximal His ligand. We found that the distal Asp-Ile hydrogen bond is important for the stability of the heme architecture and its alteration changes markedly the proximal His-Asp hydrogen-bond interaction. PMID- 15134454 TI - Functional properties of recombinant factor V mutated in a potential calcium binding site. AB - Activated coagulation factor V (FVa) is a cofactor of activated factor X (FXa) in prothrombin activation. FVa is composed of a light chain (LC) and a heavy chain (HC) that are noncovalently associated in a calcium-dependent manner. We constructed a recombinant FV Asp111Asn/Asp112Asn mutant (rFV-NN) to abolish calcium binding to a potential calcium-binding site in FVa in order to study the specific role of these residues in the expression of FVa activity. Whereas thrombin-activated recombinant FV wild type (rFV-wt) presented with stable FVa activity, incubation of rFV-NN with thrombin resulted in a temporary increase in FVa activity, which was rapidly lost upon prolonged incubation. Loss of FVa activity was most likely due to dissociation of HC and LC since, upon chromatography of rFVa-NN on a SP-Sepharose column, the HC did not bind significantly to the resin whereas the LC bound and could be eluted at high ionic strength. In contrast, rFVa-wt adhered to the column, and both the HC and LC coeluted at high ionic strength. In the presence of phospholipid vesicles, the loss of rFVa-NN activity was partially prevented by FXa, active site inhibited FXa, and prothombin in a dose-dependent manner. We conclude that the introduced amino acid substitutions result in a loss of the high-affinity (calcium dependent) interaction of the HC and LC of FVa. We propose that the introduced substitutions disrupt the calcium-binding site in FV, thereby yielding a FV molecule that rapidly loses activity following thrombin-catalyzed activation most likely via dissociation of the HC and LC. PMID- 15134455 TI - Kinetics and mechanism of iron(III) complexation by ferric binding protein: the role of phosphate. AB - Iron transport across the periplasmic space to the cytoplasmic membrane of certain Gram-negative bacteria is mediated by a ferric binding protein (Fbp). This requires Fe(3+) loading of Fbp at the inner leaflet of the outer membrane. A synergistic anion is required for tight Fe(3+) sequestration by Fbp. Although phosphate fills this role in the protein isolated from bacterial cell lysates, nitrilotriacetate anion (NTA) can also satisfy this requirement in vitro. Here, we report the kinetics and mechanism of Fe(3+) loading of Fbp from Fe(NTA)(aq) in the presence of phosphate at pH 6.5. The reaction proceeds in four kinetically distinguishable steps to produce Fe(3+)Fbp(PO(4)) as a final product. The first three steps exhibit half-lives ranging from ca. 20 ms to 0.5 min, depending on the concentrations, and produce Fe(3+)Fbp(NTA) as an intermediate product of significant stability. The rate for the first step is accelerated with an increasing phosphate concentration, while that of the third step is retarded by phosphate. Conversion of Fe(3+)Fbp(NTA) to Fe(3+)Fbp(PO(4)) in the fourth step is a slow process (half-life approximately 2 h) and is facilitated by free phosphate. A mechanism for the Fe(3+)-loading process is proposed in which the synergistic anions, phosphate and NTA, play key roles. These data suggest that not only is a synergistic anion required for tight Fe(3+) sequestration by Fbp, but also the synergistic anion plays a critical role in the process of inserting Fe(3+) into the Fbp binding site. PMID- 15134456 TI - Long-range nature of the interactions between titratable groups in Bacillus agaradhaerens family 11 xylanase: pH titration of B. agaradhaerens xylanase. AB - Xylanase from Bacillus agaradhaerens belongs to a large group of glycosyl hydrolases which catalyze the degradation of xylan. The protonation behavior of titratable groups of the uniformly (15)N- and (13)C-labeled xylanase was investigated by multinuclear NMR spectroscopy. A total of 224 chemical shift titration curves corresponding to (1)H, (13)C, and (15)N resonances revealed pK(a) values for all aspartic and glutamic acid residues, as well as for the C terminal carboxylate and histidine residues. Most of the titratable groups exhibit a complex titration behavior, which is most likely due to the mutual interactions with other neighboring groups or due to an unusual local microenvironment. Subsite -1 containing the catalytic dyad shows a long-range interaction over 9 A with Asp21 via two hydrogen bonds with Asn45 as the mediator. This result illuminates the pivotal role of the conserved position 45 among family 11 endoxylanases, determining an alkaline pH optimum by asparagine residues or an acidic pH optimum by an aspartate. The asymmetric interactions of neighboring tryptophan side chains with respect to the catalytic dyad can be comprehended as a result of hydrogen bonding and aromatic stacking. Most of the chemical shift-pH profiles of the backbone amides exhibit biphasic behavior with two distinct inflection points, which correspond to the pK(a) values of the nearby acidic side chains. However, the alternation of both positive and negative slopes of individual amide titration curves is interpreted as a consequence of a simultaneous reorganization of side chain conformational space at pH approximately 6 and/or an overall change in the hydrogen network in the substrate binding cleft. PMID- 15134457 TI - Alanine scanning mutagenesis of the testosterone binding site of rat 3 alpha hydroxysteroid dehydrogenase demonstrates contact residues influence the rate determining step. AB - Aldo-keto reductase (AKR1C) isoforms can regulate ligand access to nuclear receptors by acting as hydroxysteroid dehydrogenases. The principles that govern steroid hormone binding and steroid turnover by these enzymes were analyzed using rat 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD, AKR1C9) as the protein model. Systematic alanine scanning mutagenesis was performed on the substrate binding pocket as defined by the crystal structure of the 3alpha HSD.NADP(+).testosterone ternary complex. T24, L54, F118, F129, T226, W227, N306, and Y310 were individually mutated to alanine, while catalytic residues Y55 and H117 were unaltered. The effects of these mutations on the ordered bi-bi mechanism were examined. No mutations changed the affinity for NADPH by more than 2-3-fold. Fluorescence titrations of the energy transfer band of the E.NADPH complex with competitive inhibitors testosterone and progesterone showed that the largest effect was a 23-fold decrease in the affinity for progesterone in the W227A mutant. By contrast, changes in the K(d) for testosterone were negligible. Examination of the k(cat)/K(m) data for these mutants indicated that, irrespective of steroid substrate, the bimolecular rate constant was more adversely affected when alanine replaced an aromatic hydrophobic residue. By far, the greatest effects were on k(cat) (decreases of more than 2 log units), suggesting that the rate-determining step was either altered or slowed significantly. Single- and multiple-turnover experiments for androsterone oxidation showed that while the wild-type enzyme demonstrated a k(lim) and burst kinetics consistent with slow product release, the W227A and F118A mutants eliminated this kinetic profile. Instead, single- and multiple-turnover experiments gave k(lim) and k(max) values identical with k(cat) values, respectively, indicating that chemistry was now rate-limiting overall. Thus, conserved residues within the steroid-binding pocket affect k(cat) more than K(d) by influencing the rate-determining step of steroid oxidation. These findings support the concept of enzyme catalysis in which the correct positioning of reactants is essential; otherwise, k(cat) will be limited by the chemical event. PMID- 15134459 TI - Oligosaccharides implicated in recognition are predicted to have relatively ordered structures. AB - Fucosylated O- and N-linked glycans are essential recognition molecules in plants and animals. To understand how they impart their functions, through interactions with proteins, requires a detailed analysis of structure and dynamics, but this is presently lacking. In this study, the three-dimensional structure and dynamics of three fucosylated oligosaccharides are investigated using a combination of high field (800 MHz) nuclear magnetic resonance and long (50 ns) molecular dynamics simulations in explicit water. Predictions from dynamics simulations were in agreement with nuclear Overhauser cross-peak intensities. Similarly, a theory of weak alignment in neutral media resulted in reasonable predictions of residual dipolar couplings for the trisaccharide fucosyllactose. However, for larger penta- and hexasaccharides (LNF-1 and LND-1), the anisotropic component of the alignment was underestimated, attributed to shape irregularities of the fucosyl branches on an otherwise linear core, being more pronounced in a singly branched than a doubly branched oligosaccharide. Simulations, confirmed by experiment, predicted fucosylated molecules that are restricted to librations about a single average conformation. This restriction is partly due to microscopic water interactions, which act to stabilize intramolecular hydrogen bonds and maintain tight and ordered conformations; a view not forthcoming from simpler, nonaqueous simulations. Such a conclusion is crucial for understanding how these molecules interact with proteins and impart their recognition properties. PMID- 15134458 TI - Electron paramagnetic resonance reveals a large-scale conformational change in the cytoplasmic domain of phospholamban upon binding to the sarcoplasmic reticulum Ca-ATPase. AB - We used EPR spectroscopy to probe directly the interaction between phospholamban (PLB) and its regulatory target, the sarcoplasmic reticulum Ca-ATPase (SERCA). Synthetic monomeric PLB was prepared with a single cytoplasmic cysteine at residue 11, which was then spin labeled. PLB was reconstituted into membranes in the presence or absence of SERCA, and spin label mobility and accessibility were measured. The spin label was quite rotationally mobile in the absence of SERCA, but became more restricted in the presence of SERCA. SERCA also decreased the dependence of spin label mobility on PLB concentration in the membrane, indicating that SERCA reduces PLB-PLB interactions. The spin label MTSSL, attached to Cys11 on PLB by a disulfide bond, was stable at position 11 in the absence of SERCA. In the presence of SERCA, the spin label was released and a covalent bond was formed between PLB and SERCA, indicating direct interaction of one or more SERCA cysteine residues with Cys11 on PLB. The accessibility of the PLB-bound spin label IPSL to paramagnetic agents, localized in different phases of the membrane, indicates that SERCA greatly reduces the level of interaction of the spin label with the membrane surface. We propose that the cytoplasmic domain of PLB associates with the lipid surface, and that association with SERCA induces a major conformational change in PLB in which the cytoplasmic domain is drawn away from the lipid surface by SERCA. PMID- 15134460 TI - Short sequences of non-proline residues can adopt the polyproline II helical conformation. AB - The left-handed polyproline II (P(II)) helix is a structure that has been given a great deal of attention lately because of its role in a wide variety of physiologically important processes and potential significance in protein unfolded states. Recent work by several authors has shown that residues besides proline can adopt this structure. A scale of relative P(II)-helix-forming propensities has been generated but only for single guest residues in a proline based host system. Here, we present multiple guest residues in a proline-based host system. Using circular dichroism spectroscopy, we have shown that not only single residues, but also short sequences of non-proline residues can adopt the P(II) conformation. PMID- 15134462 TI - Structure-function relationship of the influenza virus RNA polymerase: primer binding site on the PB1 subunit. AB - Influenza virus RNA polymerase is composed of three viral P proteins (PB1, PB2, and PA) and involved in both transcription and replication of the viral RNA genome. The catalytic site for RNA polymerization is located on the PB1 subunit. To identify the primer ATP-binding site, we have employed a highly selective cross-linking technique: three structurally diverse ATP analogues with reactive groups on their phosphate moieties were first cross-linked to the viral RNA polymerase, and the cross-linked nucleotides were then elongated to dinucleotides by adding the second substrate [alpha-(32)P]GTP. Only the ATP analogues tethered to the primer-binding site could be elongated to radioactive AG dinucleotides. Using this catalytically competent cross-linking procedure, the PB1 subunit was found to be the only labeled subunit. Limited proteolysis of the labeled PB1 by V8 protease revealed the segment between amino acids 179 and 297 as the only bearer of the radioactive label. Thus, we concluded that this region of PB1 faces the 5' end of the primer nucleotide. In support of this prediction, the cross linked dinucleotides were further elongated up to 8-10 nucleotides in length upon addition of all four substrates. This result suggests that the substantial mass of RNA can be accommodated between the binding site for the primer (and nascent RNA) and the catalytic center of RNA polymerization. PMID- 15134461 TI - Entropy-driven folding of an RNA helical junction: an isothermal titration calorimetric analysis of the hammerhead ribozyme. AB - Helical junctions are extremely common motifs in naturally occurring RNAs, but little is known about the thermodynamics that drive their folding. Studies of junction folding face several challenges: non-two-state folding behavior, superposition of secondary and tertiary structural energetics, and drastically opposing enthalpic and entropic contributions to folding. Here we describe a thermodynamic dissection of the folding of the hammerhead ribozyme, a three-way RNA helical junction, by using isothermal titration calorimetry of bimolecular RNA constructs. By using this method, we show that tertiary folding of the hammerhead core occurs with a highly unfavorable enthalpy change, and is therefore entropically driven. Furthermore, the enthalpies and heat capacities of core folding are the same whether supported by monovalent or divalent ions. These properties appear to be general to the core sequence of bimolecular hammerhead constructs. We present a model for the ion-induced folding of the hammerhead core that is similar to those advanced for the folding of much larger RNAs, involving ion-induced collapse to a structured, non-native state accompanied by rearrangement of core residues to produce the native fold. In agreement with previous enzymological and structural studies, our thermodynamic data suggest that the hammerhead structure is stabilized in vitro predominantly by diffusely bound ions. Our approach addresses several significant challenges that accompany the study of junction folding, and should prove useful in defining the thermodynamic determinants of stability in these important RNA motifs. PMID- 15134464 TI - Full-length influenza hemagglutinin HA2 refolds into the trimeric low-pH-induced conformation. AB - The influenza virus uses hemagglutinin (HA) to fuse the viral and cellular membranes. As part of an effort to study the membrane-interacting elements of HA, the fusion peptide, and the C-terminal transmembrane anchor, we have expressed in Escherichia coli the full-length HA(2) chain with maltose-binding protein fused at its N-terminus. The chimeric protein can be refolded in vitro in the presence of specific detergents to yield stable, homogeneous trimers, as determined by analytical ultracentrifugation. The trimers have the so-called "low pH" conformation-the rearranged HA(2) conformation obtained when intact HA(1)/HA(2) is induced to refold by exposure to low pH-as detected by electron microscopy and monoclonalantibody reactivity. These results provide further evidence for the notion that the neutral-pH structure of intact HA is metastable and that binding of protons lowers the kinetic barriers that prevent rearrangement to the minimum free-energy conformation. The refolded chimeric protein described here is a suitable species for undertaking studies of how the fusion peptide inserts into membranes and assessing the nature of possible intermediates in the fusion process. PMID- 15134463 TI - In vitro phosphorylation of insulin receptor substrate 1 by protein kinase C zeta: functional analysis and identification of novel phosphorylation sites. AB - Protein kinase C-zeta (PKC-zeta) participates both in downstream insulin signaling and in the negative feedback control of insulin action. Here we used an in vitro approach to identify PKC-zeta phosphorylation sites within insulin receptor substrate 1 (IRS-1) and to characterize the functional implications. A recombinant IRS-1 fragment (rIRS-1(449)(-)(664)) containing major tyrosine motifs for interaction with phosphatidylinositol (PI) 3-kinase strongly associated to the p85alpha subunit of PI 3-kinase after Tyr phosphorylation by the insulin receptor. Phosphorylation of rIRS-1(449)(-)(664) by PKC-zeta induced a prominent inhibition of this process with a mixture of classical PKC isoforms being less effective. Both PKC-zeta and the classical isoforms phosphorylated rIRS-1(449)( )(664) on Ser(612). However, modification of this residue did not reduce the affinity of p85alpha binding to pTyr-containing peptides (amino acids 605-615 of rat IRS-1), as determined by surface plasmon resonance. rIRS-1(449)(-)(664) was then phosphorylated by PKC-zeta using [(32)P]ATP and subjected to tryptic phosphopeptide mapping based on two-dimensional HPLC coupled to mass spectrometry. Ser(498) and Ser(570) were identified as novel phosphoserine sites targeted by PKC-zeta. Both sites were additionally confirmed by phosphopeptide mapping of the corresponding Ser --> Ala mutants of rIRS-1(449)(-)(664). Ser(570) was specifically targeted by PKC-zeta, as shown by immunoblotting with a phosphospecific antiserum against Ser(570) of IRS-1. Binding of p85alpha to the S570A mutant was less susceptible to inhibition by PKC-zeta, when compared to the S612A mutant. In conclusion, our in vitro data demonstrate a strong inhibitory action of PKC-zeta at the level of IRS-1/PI 3-kinase interaction involving multiple serine phosphorylation sites. Whereas Ser(612) appears not to participate in the negative control of insulin signaling, Ser(570) may at least partly contribute to this process. PMID- 15134465 TI - Identification of the catalytic residues involved in the carboxyl transfer of pyruvate carboxylase. AB - To clarify the mechanism of carboxyl transfer from carboxylbiotin to pyruvate, the following conserved amino acid residues present in the carboxyl transferase domain of Bacillus thermodenitrificans pyruvate carboxylase were converted to homologous amino acids: Asp543, Glu576, Glu592, Asp649, Lys712, Asp713, and Asp762. The carboxylase activity of the resulting mutants, D543E, E576D, E576Q, E592Q, D649N, K712R, K712Q, D713E, D713N, D762E, and D762N, was generally less than that of the wild type from mutation, but it decreased the most to 5% or even less than that of the wild type with D543E, D576Q, D649N, K712R, and K712Q. The decrease in activity observed for Asp543, Asp649, and Lys712 mutants was not for structural reasons because their structures seemed to remain intact as assessed by gel filtration and circular dichroism. On the basis of these data, a mechanism is proposed where Lys712 and Asp543 serve as the key acid and base catalyst, respectively. PMID- 15134466 TI - Hypercoagulability in renal transplant recipients. Identifying patients at risk of renal allograft thrombosis and evaluating strategies for prevention. AB - Renal transplantation improves survival and quality of life for patients with end stage renal disease (ESRD). Improvements in immunosuppressive therapy have reduced early allograft loss due to acute rejection to very low levels. Early allograft loss, due to acute thrombotic complications, remains a constant and proportionally increasing complication of renal transplantation. Identifying risk factor(s) for thrombosis amenable to preventive strategies has been elusive. Epidemiological studies have attempted to define risk in terms of modifiable (drugs, dialysis modality, surgical procedure) and non-modifiable (age, diabetes mellitus, vascular anomalies) factors, or identify changes in coagulation or fibrinolysis promoting a more thrombotic state. Most recently the evolution of thrombophilia research has established the potential for inherited hypercoagulability to predispose to acute allograft thrombosis. Inheritance of the factor V Leiden (FVL), prothrombin G20210A mutation, or the presence of antiphospholipid antibodies (APA) may increase the risk of renal allograft thrombosis approximately 3-fold in selected patients. Patients with ESRD due to systemic lupus erythematosus (SLE) appear at particularly high risk of thrombosis, especially if they have either APA or detectable beta(2)-glycoprotein 1. Data for other hypercoagulable states such as hyperhomocystinemia or the C677T polymorphism of the methylenetetrahydrofolate reductase gene are deficient. Patients with APA, FVL, or prothrombin G20210A mutation also appear to have greater graft loss due to vascular rejection, possibly reflecting immunological injury upon the vascular wall exacerbated or induced by the prothrombotic state. While substantial in vitro data suggest cyclosporine is prothrombotic, an independent clinical association with allograft thrombosis is unproven. Interventions to reduce thrombotic risk including heparin, warfarin, and aspirin have been evaluated in both selected high-risk groups (heparin and warfarin) and unselected populations (heparin and aspirin). In unselected patients at low clinical risk, aspirin (75-150 mg/day) with or without a short period of unfractionated heparin (5000U twice a day for 5 days) appears to reduce the risk of renal allograft thrombosis significantly with a low risk of bleeding, especially when compared with low molecular weight heparins which risk accumulation in renal failure. In high-risk groups (identified thrombophilic risk factor, previous thrombosis, or SLE) longer period of heparin, with or without aspirin and maintenance with warfarin, should be considered. Re-transplantation following graft loss due to vascular thrombosis can be undertaken with a low risk of recurrence. Further prospective studies evaluating both putative risk factors and intervention strategies are required to determine whether routine clinical screening for thrombophilic factors is justified. PMID- 15134467 TI - Aspirin in cardiovascular disorders. What is the optimum dose? AB - Clinical trials of aspirin (acetylsalicylic acid) for cardiovascular disorders have employed doses defined for other pharmacological effects of the drug (such as analgesic effects). Antioxidant and anti-inflammatory mechanisms with different dose-response relationships may contribute to the clinical effect of aspirin in cardiovascular disease. The optimal aspirin dose remains uncertain. Although the difference between 325 mg/day and 81 mg/day of aspirin sounds trivial, finding an optimal aspirin dose has enormous potential to reduce ischemic events. Large aspirin doses have not been associated with proportionally greater benefit. For patients with ischemic heart disease, overall consensus defines a range between 75 and 160 mg/day for the secondary prevention of myocardial infarction, stroke, and vascular death. Any benefit of aspirin must be measured against its adverse effects, principally gastrointestinal hemorrhage. The potential for adverse bleeding events may be lower with a 81mg dose, while maintaining clinical benefit. Although current aggregate data is reassuring about aspirin administration, it is increasingly clear that existing aspirin studies are insufficient to conclusively determine an optimal aspirin dose. Platelets can be activated by pathways that are not blocked by aspirin, and the dose of aspirin needed to fully suppress platelet aggregation may be higher in some patients as a result. Higher doses of aspirin than are currently used (75-325 mg/day) may be required in these patients to achieve desired antithrombotic effects. Better understanding of aspirin-resistant populations will facilitate identification of patients who require higher aspirin doses or alternative forms of antiplatelet therapy. PMID- 15134468 TI - Pharmacological prevention of reperfusion injury in acute myocardial infarction. A potential role for adenosine as a therapeutic agent. AB - The concept of reperfusion injury, although first recognized from animal studies, is now recognized as a clinical phenomenon that may result in microvascular damage, no-reflow phenomenon, myocardial stunning, myocardial hibernation and ischemic preconditioning. The final consequence of this event is left ventricular (LV) systolic dysfunction leading to increased morbidity and mortality. The typical clinical case of reperfusion injury occurs in acute myocardial infarction (MI) with ST segment elevation in which an occlusion of a major epicardial coronary artery is followed by recanalization of the artery. This may occur either spontaneously or by means of thrombolysis and/or by primary percutaneous coronary intervention (PCI) with efficient platelet inhibition by aspirin (acetylsalicylic acid), clopidogrel and glycoprotein IIb/IIIa inhibitors. Although the pathophysiology of reperfusion injury is complex, the major role that neutrophils play in this process is well known. Neutrophils generate free radicals, degranulation products, arachidonic acid metabolites and platelet activating factors that interact with endothelial cells, inducing endothelial injury and neutralization of nitrous oxide vasodilator capacity. Adenosine, through its multi-targeted pharmacological actions, is able to inhibit some of the above-mentioned detrimental effects. The net protective of adenosine in in vivo models of reperfusion injury is the reduction of the infarct size, the improvement of the regional myocardial blood flow and of the regional function of the ischemic area. Additionally, adenosine preserves the post-ischemic coronary flow reserve, coronary blood flow and the post-ischemic regional contractility. In small-scale studies in patients with acute MI, treatment with adenosine has been associated with smaller infarcts, less no-reflow phenomenon and improved LV function. During elective PCI adenosine reduced ST segment shifts, lactate production and ischemic symptoms. During the last years, three relatively large placebo-controlled clinical trials have been conducted: Acute Myocardial Infarction Study of Adenosine Trial (AMISTAD) I and II and Attenuation by Adenosine of Cardiac Complications (ATTACC). In the AMISTAD trials, the final infarct size was reduced and the LV systolic function was improved by adenosine treatment, mainly in patients with anterior MI localization. However, morbidity and mortality were not affected. In the ATTACC study, the LV systolic function was not affected by adenosine, however, trends towards improved survival were observed in patients with anterior MI localization. The possibility of obtaining a Thrombolysis in Myocardial Infarction (TIMI) grade 3 flow in the infarct related artery in up to 95% of patients with acute MI (increasing the occurrence of reperfusion injury) has turned back the interest towards the protection of myocardial cells from the impending ischemic and reperfusion injury in which adenosine alone or together with other cardio-protective agents may exert important clinical effects. PMID- 15134469 TI - Therapeutic potential of anticytokine therapy in congestive heart failure. AB - Accumulating evidence indicates that inflammatory cytokines play a pathogenic role in congestive heart failure (CHF) by influencing heart contractility, inducing hypertrophy, and promoting apoptosis or fibrosis, contributing to the continuous myocardial remodeling process. While several stimuli may be operating such as heat-shock protein, microbial antigen, bacterial lipopolysaccharide, shear and oxidative stress, hypoxia and oxidized low-density lipoprotein (LDL), it seems that the inflammatory response to these stimuli may represent a common final pathogenic pathway in CHF regardless of the initial event. Traditional cardiovascular drugs seem to have little influence on the overall cytokine network, and immunomudulatory therapy has emerged as a possible new treatment modality in CHF. Several animal studies, and some clinical pilot studies, have suggested that down-regulation of inflammatory cytokines may improve cardiac performance. On the other hand, preliminary results from the placebo-controlled studies suggest no effect, or even adverse effect, of antitumor necrosis factor (TNF) therapy on mortality and hospitalization. Although somewhat disappointing, these negative results do not necessarily argue against the 'cytokine hypothesis'. These studies just underscore the difficulties and the challenges in developing treatment modalities that can modulate the cytokine network in CHF patients resulting in anti-inflammatory and beneficial net effects. Further research in this area will have to more precisely identify the most important actors in the immunopathogenesis of CHF in order to develop more specific immunomodulating agents for this disorder. However, at present the beneficial role of anticytokine therapy in patients with CHF remains unproven. PMID- 15134470 TI - Cardiac syndrome X. Diagnosis, pathogenesis and management. AB - Patients with cardiac syndrome X (typical chest pain and normal coronary arteriograms) represent a heterogeneous syndrome, which encompasses different pathogenic mechanisms. Although symptoms in most patients with cardiac syndrome X are non-cardiac, a sizable proportion of them have angina pectoris due to transient myocardial ischemia. Thus radionuclide myocardial perfusion defects, coronary sinus oxygen saturation abnormalities and pH changes, myocardial lactate production and stress-induced alterations of cardiac high energy phosphate suggest an ischemic origin of symptoms in at least a proportion of patients with cardiac syndrome X. Microvascular abnormalities, caused by endothelial dysfunction, appear to be responsible for myocardial ischemia in patients with cardiac syndrome X. Endothelial dysfunction is likely to be multifactorial in these patients and it is conceivable that risk factors such as hypertension, hypercholesterolemia, diabetes mellitus and smoking can contribute to its development. Most patients with cardiac syndrome X are postmenopausal women and estrogen deficiency has been therefore proposed as a pathogenic factor in female patients. Additional factors such as abnormal pain perception may contribute to the pathogenesis of chest pain in patients with angina pectoris and normal coronary angiograms. Although prognosis is good regarding survival, patients with cardiac syndrome X have an impaired quality of life. Management of this syndrome represents a major challenge to the treating physician. Understanding the mechanism underlying the condition is of vital importance for patient management. Thus diagnostic tests should aim at identifying the cause of the symptoms in the individual patient, i.e. myocardial ischemia, increased pain perception, abnormalities of adrenergic tone, non-cardiac mechanisms, etc. Moreover, it is important to bear in mind that treatment of cardiac syndrome X should be mainly directed towards improving quality of life, as prognosis is usually good in these patients. Conventional antianginal agents such nitrates, calcium channel antagonists, beta-adrenoceptor antagonists and nicorandil are effective particularly in patients in whom chest pain and ECG changes are clearly suggestive of myocardial ischemia and in those with objective documentation of ischemia. Angiotensin-converting enzyme inhibitors have been shown to be useful in syndrome X patients with increased adrenergic tone, borderline systemic hypertension, and those with documented endothelial dysfunction. Analgesic interventions of different sorts have been proposed based on the hypothesis that somatic and visceral perception of pain is altered in cardiac syndrome X patients. Pharmacological agents such as imipramine and aminophylline, and neural electrical stimulation techniques have been assessed in recent years with encouraging results. Psychological treatment, particularly cognitive therapy, appears to be useful in defined patient subsets. Relaxation techniques such as transcendental meditation have been successfully used in small studies and shown to improve not only chest pain but also exercise-induced ST segment changes. Reports indicate that these techniques improve quality of life. PMID- 15134471 TI - Efficacy and tolerability of long-term rilmenidine treatment in hypertensive diabetic patients. A retrospective analysis of a general practice study. AB - INTRODUCTION: Rilmenidine is a centrally acting antihypertensive which differs from the other representatives of this class by its very high specificity for the imidazoline I1 receptors and its good tolerability. Recent studies have shown rilmenidine improves glucose tolerance and reduces micro-albuminuria in patients with diabetes mellitus. METHODOLOGY: The evidence of these potentially favorable characteristics encouraged a secondary retrospective analysis of a subgroup of 2738 diabetic patients included in a previous long-term open study of rilmenidine alone, or in combination with other classes of antihypertensives. RESULTS: The antihypertensive efficacy of rilmenidine demonstrated previously in controlled studies was confirmed during the 12-month follow-up. In addition, favorable effects of drug treatment on fasting blood glucose and plasma triglyceride levels were consistent with an improvement in glucose and lipid metabolism during treatment. The profile of adverse events was similar to that observed in the nondiabetic population, the occurrence of postural hypotension being observed in <1% of patients and not necessitating any withdrawals from the study. CONCLUSION: Diabetic hypertensive patients frequently require the use of multiple medications and consideration of the metabolic interactions between treatments. The results of this retrospective analysis support the use of rilmenidine in patients with hypertension and diabetes mellitus and should encourage the conduct of controlled trials of cardiovascular and renal protection and outcome with this compound. PMID- 15134472 TI - Wave aberrations of the isolated crystalline lens. AB - A method to measure wave aberrations in the isolated crystalline lens is demonstrated. The method employs a laser scanning technique in which the trajectories of narrow refracted laser beams are measured for an array of sample positions incident on the lens. The local slope of the emerging wavefront is calculated for each sample position, and a least squares procedure is used to fit a Zernike polynomial function to define the wave aberration. Measurements of the aberrations of an isolated porcine lens and macaque lens undergoing changes in accommodative state with mechanical stretching are shown. Many aberrations were present, but negative spherical aberration dominated. In the macaque lens, many aberrations underwent systematic changes with accommodation, most notably the 4th order spherical aberration, which became more negative, and the 6th order spherical aberration, which progressed from negative to positive. PMID- 15134473 TI - Compensation of corneal horizontal/vertical astigmatism, lateral coma, and spherical aberration by internal optics of the eye. AB - Both the anterior surface of the cornea and the internal optics (the posterior cornea, crystalline lens) contribute to the aberration of a wavefront passing through the eye. Artal, Guirao, Berrio, and Williams (2001) reported that the wavefront aberrations produced by the internal optics offset, or compensate for, the aberrations produced by the cornea to reduce ocular wavefront aberrations. We have investigated the wavefront aberrations of the cornea, internal optics, and complete eye on both the population and individual level to determine which aberrations are compensated and probable paths leading to that compensation. The corneal and ocular aberrations of 30 young subjects at relaxed accommodation were measured with the Topcon Wavefront Analyzer, which simultaneously measures refraction, corneal topography (videokeratoscope), and wavefront aberrations (Hartmann-Shack sensor). We found strong evidence for compensation of horizontal/vertical (H/V) astigmatism (Zernike term Z5) lateral coma (Z8) and spherical aberration (Z12). H/V astigmatism compensation is scaled for each individual, suggesting that it is actively determined by a fine-tuning process. Spherical aberration shows no individual compensation, suggesting that is a passive result of genetically determined physiology. Lateral coma shows individually scaled compensation, some of which may be attributable to eccentricity of the fovea. PMID- 15134474 TI - A population study on changes in wave aberrations with accommodation. AB - Wave aberrations were measured with a Shack-Hartmann wavefront sensor (SHWS) in the right eye of a large young adult population when accommodative demands of 0, 3, and 6 D were presented to the tested eye through a Badal system. Three SHWS images were recorded at each accommodative demand and wave aberrations were computed over a 5-mm pupil (through 6th order Zernike polynomials). The accommodative response was calculated from the Zernike defocus over the central 3 mm diameter zone. Among all individual Zernike terms, spherical aberration showed the greatest change with accommodation. The change of spherical aberration was always negative, and was proportional to the change in accommodative response. Coma and astigmatism also changed with accommodation, but the direction of the change was variable. Despite the large inter-subject variability, the population average of the root mean square for all aberrations (excluding defocus) remained constant for accommodative levels up to 3.0 D. Even though aberrations change with accommodation, the magnitude of the aberration change remains less than the magnitude of the uncorrected aberrations, even at high accommodative levels. Therefore, a typical eye will benefit over the entire accommodative range (0-6 D) if aberrations are corrected for distance viewing. PMID- 15134475 TI - Neural compensation for the eye's optical aberrations. AB - A fundamental problem facing sensory systems is to recover useful information about the external world from signals that are corrupted by the sensory process itself. Retinal images in the human eye are affected by optical aberrations that cannot be corrected with ordinary spectacles or contact lenses, and the specific pattern of these aberrations is different in every eye. Though these aberrations always blur the retinal image, our subjective impression is that the visual world is sharp and clear, suggesting that the brain might compensate for their subjective influence. The recent introduction of adaptive optics to control the eye's aberrations now makes it possible to directly test this idea. If the brain compensates for the eye's aberrations, vision should be clearest with the eye's own aberrations rather than with unfamiliar ones. We asked subjects to view a stimulus through an adaptive optics system that either recreated their own aberrations or a rotated version of them. For all five subjects tested, the stimulus seen with the subject's own aberrations was always sharper than when seen through the rotated version. This supports the hypothesis that the neural visual system is adapted to the eye's aberrations, thereby removing somehow the effects of blur generated by the sensory apparatus from visual experience. This result could have important implications for methods to correct higher order aberrations with customized refractive surgery because some benefits of optimizing the correction optically might be undone by the nervous system's compensation for the old aberrations. PMID- 15134476 TI - Myopic versus hyperopic eyes: axial length, corneal shape and optical aberrations. AB - This study investigated differences in geometrical properties and optical aberrations between a group of hyperopes and myopes (age-matched 30.3+/-5.2 and 30.5+/-3.8 years old, respectively, and with similar absolute refractive error 3.0+/-2.0 and -3.3+/-2.0, respectively). Axial length (AL) was measured by means of optical biometry, and corneal apical radius of curvature (CR) and asphericity (Q) were measured by fitting corneal topography data to biconic surfaces. Corneal aberrations were estimated from corneal topography by means of virtual ray tracing, and total aberrations were measured using a laser ray tracing technique. Internal aberrations were estimated by subtracting corneal from total aberrations. AL was significantly higher in myopes than in hyperopes and AL/CR was highly correlated with spherical equivalent. Hyperopic eyes tended to have higher (less negative) Q and higher total and corneal spherical aberration than myopic eyes. RMS for third-order aberrations was also significantly higher for the hyperopic eyes. Internal aberrations were not significantly different between the myopic and hyperopic groups, although internal spherical aberration showed a significant age-related shift toward less negative values in the hyperopic group. For these age and refraction ranges, our cross-sectional results do not support evidence of relationships between emmetropization and ocular aberrations. Our results may be indicative of presbyopic changes occurring earlier in hyperopes than in myopes. PMID- 15134477 TI - Spatially variant changes in lens power during ocular accommodation in a rhesus monkey eye. AB - This study investigated the changes in ocular aberrations that occur over the entire lens equatorial diameter during accommodation in iridectomized rhesus monkey eyes to understand the nature of accommodative lenticular deformation. Accommodation was centrally stimulated to a range of different response amplitudes (0 D to approximately 11 D), and ocular aberrations were measured with a Shack-Hartmann wavefront sensor in both eyes of one previously iridectomized 10 year-old rhesus monkey. At the highest amplitude in the two eyes, aberrations were analyzed over entrance pupil diameters ranging from 3 to 8 mm in steps of 1 mm. Root mean square error of the total measured aberrations, excluding defocus, increased systematically with increasing accommodation from about 1 to 3.5 microns. Spherical aberration became systematically more negative, and vertical coma increased significantly in magnitude with accommodation. There was a strong accommodative change in power near the center of the lens and little change in power at the periphery. At the highest accommodative state, decreasing the analyzed entrance pupil diameter from 8 to 3 mm considerably reduced the wavefront error. The greater increase in optical power near the central region of the lens, combined with an accommodative pupillary miosis, would serve to maximize accommodative refractive change while maintaining acceptable image quality. PMID- 15134478 TI - Predicting subjective judgment of best focus with objective image quality metrics. AB - PURPOSE: To determine the impact of higher-order monochromatic aberrations on lower-order subjective sphero-cylindrical refractions. METHODS: Computationally aberrated, monochromatic Sloan letters were presented on a high luminance display that was viewed by an observer through a 2.5mm pupil. Through-focus visual acuity (VA) was determined in the presence of spherical aberration (Z40) at three levels (0.10, 0.21 and 0.50D). Analogous through-astigmatism experiments measured visual acuity in the presence of secondary astigmatism (Z4+/-2) or coma (Z3-1). Measured visual acuity was correlated with 31 different metrics of image quality to determine which metric best predicts performance for degraded retinal images. The defocus and astigmatism levels that optimized each metric were compared with those that produced best visual acuity to determine which metric best predicts subjective refraction. RESULTS: Spherical aberration, coma and secondary astigmatism all reduced VA and increased depth of focus. The levels of defocus and primary astigmatism that produced the best performance varied with levels of spherical aberration and secondary astigmatism, respectively. The presence of coma, however, did not affect cylindrical refraction. Image plane metrics, especially those that take into account the neural contrast sensitivity threshold (e.g. the visual Strehl ratio, VSOTF), are good predictors of visual acuity in both the through-focus and through-astigmatism experiments (R = -0.822 for VSOTF). Subjective sphero-cylindrical refractions were accurately predicted by some image-quality metrics (e.g., pupil fraction, VSOTF and standard deviation of PSF light distribution). CONCLUSION: Subjective judgment of best focus does not minimize RMS wavefront error (Zernike defocus = 0), nor create paraxial focus (Seidel defocus = 0), but makes the retina conjugate to a plane between these two. It is possible to precisely predict subjective sphero-cylindrical refraction for monochromatic light using objective metrics. PMID- 15134479 TI - Metrics of optical quality derived from wave aberrations predict visual performance. AB - Wavefront-guided refractive surgery and custom optical corrections have reduced the residual root mean squared (RMS) wavefront error in the eye to relatively low levels (typically on the order of 0.25 microm or less over a 6-mm pupil, a dioptric equivalent of 0.19 D). It has been shown that experimental variation of the distribution of 0.25 microm of wavefront error across the pupil can cause variation in visual acuity of two lines on a standard logMAR acuity chart. This result demonstrates the need for single-value metrics other than RMS wavefront error to quantify the effects of low levels of aberration on acuity. In this work, we present the correlation of 31 single-value metrics of optical quality to high-contrast visual acuity for 34 conditions where the RMS wavefront error was equal to 0.25 microm over a 6-mm pupil. The best metric, called the visual Strehl ratio, accounts for 81% of the variance in high-contrast logMAR acuity. PMID- 15134480 TI - Accuracy and precision of objective refraction from wavefront aberrations. AB - We determined the accuracy and precision of 33 objective methods for predicting the results of conventional, sphero-cylindrical refraction from wavefront aberrations in a large population of 200 eyes. Accuracy for predicting defocus (as specified by the population mean error of prediction) varied from -0.50 D to +0.25 D across methods. Precision of these estimates (as specified by 95% limits of agreement) ranged from 0.5 to 1.0 D. All methods except one accurately predicted astigmatism to within +/-1/8D. Precision of astigmatism predictions was typically better than precision for predicting defocus and many methods were better than 0.5D. Paraxial curvature matching of the wavefront aberration map was the most accurate method for determining the spherical equivalent error whereas least-squares fitting of the wavefront was one of the least accurate methods. We argue that this result was obtained because curvature matching is a biased method that successfully predicts the biased endpoint stipulated by conventional refractions. Five methods emerged as reasonably accurate and among the most precise. Three of these were based on pupil plane metrics and two were based on image plane metrics. We argue that the accuracy of all methods might be improved by correcting for the systematic bias reported in this study. However, caution is advised because some tasks, including conventional refraction of defocus, require a biased metric whereas other tasks, such as refraction of astigmatism, are unbiased. We conclude that objective methods of refraction based on wavefront aberration maps can accurately predict the results of subjective refraction and may be more precise. If objective refractions are more precise than subjective refractions, then wavefront methods may become the new gold standard for specifying conventional and/or optimal corrections of refractive errors. PMID- 15134482 TI - ABC transporters and the blood-brain barrier. AB - The blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB) form a very effective barrier to the free diffusion of many polar solutes into the brain. Many metabolites that are polar have their brain entry facilitated by specific inwardly-directed transport mechanisms. In general the more lipid soluble a molecule or drug is, the more readily it will tend to partition into brain tissue. However, a very significant number of lipid soluble molecules, among them many useful therapeutic drugs have lower brain permeability than would be predicted from a determination of their lipid solubility. These molecules are substrates for the ABC efflux transporters which are present in the BBB and BCSB and the activity of these transporters very efficiently removes the drug from the CNS, thus limiting brain uptake. P-glycoprotein (Pgp) was the first of these ABC transporters to be described, followed by the multidrug resistance-associated proteins (MRP) and more recently breast cancer resistance protein (BCRP). All are expressed in the BBB and BCSFB and combine to reduce the brain penetration of many drugs. This phenomenon of "multidrug resistance" is a major hurdle when it comes to the delivery of therapeutics to the brain, not to mention the problem of cancer chemotherapy in general. Therefore, the development of strategies for bypassing the influence of these ABC transporters and for the design of effective drugs that are not substrates and the development of inhibitors for the ABC transporters becomes a high imperative for the pharmaceutical industry. PMID- 15134483 TI - Anti-HIV drug distribution to the central nervous system. AB - The introduction, in 1995, of highly active antiretroviral therapy (HAART) dramatically reduced the morbidity and mortality of HIV-infected patients. However, the brain remains a site of viral replication for HIV and thus is still an important target for antiretroviral agents. Consequently, a clear understanding of how the current anti-HIV drugs reach the CNS, and interact at the level of the blood-brain barrier and blood-CSF barrier, is important if we are to maximise viral suppression and improve clinical outcome. It would also contribute to the development of new anti-HIV drugs and the identification of transport inhibitors that could be used as adjuvant therapies. In this review we focus on the role of the blood-brain and blood-CSF barriers in the delivery of the main classes of approved anti-HIV drugs. Among these groups, the CNS distribution of the nucleoside reverse transcriptase inhibitors is the best characterised. It involves probenecid efflux transport mechanisms, which limit their brain delivery and probably their, neurological efficacy. Nevirapine and efavirenz, the commonly prescribed non-nucleoside reverse transcriptase inhibitors, can readily enter the CSF, however, it remains to be seen if a transport system is involved in their distribution. The protease inhibitors have only a limited ability to reach the CNS, with the majority of this class of drugs not even being detected in human CSF after administration. This is partly the result of their removal from the CNS by the efflux transporters; P-glycoprotein, and possibly multi-drug resistance associated protein (MRP). PMID- 15134484 TI - Passage of VIP/PACAP/secretin family across the blood-brain barrier: therapeutic effects. AB - In recent years, VIP/PACAP/secretin family has special interest. Family members are vasoactive intestinal peptide (VIP), pituitary adenylate cyclase-activating polypeptide (PACAP), secretin, glucagon, glucagon like peptide-1 (GLP(1)), GLP(2), gastric inhibitory peptide (GIP), growth hormone releasing hormone (GHRH or GRF), and peptide histidine methionine (PHM). Most of the family members present both in central nervous system (CNS) and in various peripheral tissues. The family members that are released into blood from periphery, especially gut, circulate the brain and they can cross the blood brain barrier. On the other hand, some of the members of this family that present in the brain, can cross from brain to blood and reach the peripheral targets. VIP, secretin, GLP(1), and PACAP 27 are transported into the brain by transmembrane diffusion, a non saturable mechanism. However, uptake of PACAP 38 into the brain is saturable mechanism. While there is no report for the passage of GIP, GLP(2), and PHM, there is only one report that shows, glucagon and GHRH can cross the BBB. The passage of VIP/PACAP/secretin family members opens up new horizon for understanding of CNS effects of peripherally administrated peptides. There is much hope that those peptides may prove to be useful in the treatment of serious neurological diseases such as Alzheimer's disease, amyotropic lateral sclerosis, Parkinson's disease, AIDS related neuropathy, diabetic neuropathy, autism, stroke and nerve injury. Their benefits in various pathophysiologic conditions undoubtly motivate the development of a novel drug design for future therapeutics. PMID- 15134485 TI - Drug delivery systems for brain tumor therapy. AB - Brain tumors are one of the most lethal forms of cancer. They are extremely difficult to treat. Although, the rate of brain tumor incidence is relatively low, the field clearly lacks therapeutic strategies capable of overcoming barriers for effective delivery of drugs to brain tumors. Clinical failure of many potentially effective therapeutics for the treatment of brain tumors is usually not due to a lack of drug potency, but rather can be attributed to shortcomings in the methods by which a drug is delivered to the brain and into brain tumors. In response to the lack of efficacy of conventional drug delivery methods, extensive efforts have been made to develop novel strategies to overcome the obstacles for brain tumor drug delivery. The challenge is to design therapeutic strategies that deliver drugs to brain tumors in a safe and effective manner. This review provides some insight into several potential techniques that have been developed to improve drug delivery to brain tumors, and it should be helpful to clinicians and research scientists as well. PMID- 15134487 TI - Are the extracellular [correction of extracelluar] pathways a conduit for the delivery of therapeutics to the brain? AB - Most drugs with central nervous system (CNS) activity enter the brain either by diffusing across the membranes which comprise the blood-brain barrier (BBB) or by being transported by carrier systems across those membranes. Substances which cannot cross the BBB by one of these mechanisms, like serum albumin, are virtually excluded from the CNS. However, this exclusion is not absolute. Cerebrospinal fluid (CSF) levels of albumin, for example, are about 0.5% those of serum levels. Albumin enters the CNS through a variety of pathways collectively termed the extracellular pathways. Any circulating substance can, in theory, use these pathways to enter the CNS. But, traditional drug development has ignored this pathway. To approach even the CSF/serum ratio of 0.5%, a candidate therapeutic would need to meet several criterion: long half-life in blood, small volume of distribution, high potency in the CNS, and absence of brain-to-blood efflux. Two emerging therapeutics which are likely exerting their CNS effects by way of the extracellular pathways are antibodies directed against amyloid beta protein (ABP) and erythropoietin (Epo) used in the treatment of stroke. These examples suggest that the extracellular pathways are an option for the delivery of certain therapeutics to the brain. PMID- 15134486 TI - New technologies for drug delivery across the blood brain barrier. AB - The blood-brain barrier (BBB) efficiently restricts penetration of therapeutic agents to the brain from the periphery. Therefore, discovery of new modalities allowing for effective delivery of drugs and biomacromolecules to the central nervous system (CNS) is of great need and importance for treatment of neurodegenerative disorders. This manuscript focuses on three relatively new strategies. The first strategy involves inhibition of the drug efflux transporters expressed in BBB by Pluronic block copolymers, which allows for the increased transport of the substrates of these transporters to the brain. The second strategy involves the design of nanoparticles conjugated with specific ligands that can target receptors in the brain microvasculature and carry the drugs to the brain through the receptor mediated transcytosis. The third strategy involves artificial hydrophobization of peptides and proteins that facilitates the delivery of these peptides and proteins across BBB. This review discusses the current state, advantages and limitations of each of the three technologies and outlines their future prospects. PMID- 15134488 TI - Indolactam and benzolactam compounds as new medicinal leads with binding selectivity for C1 domains of protein kinase C isozymes. AB - Protein kinase C (PKC) isozymes (alpha, betaI, betaII, gamma, delta, epsilon, eta, theta) are major receptors of tumor promoters and also play a crucial role in cellular signal transduction via the second messenger, 1,2-diacyl-sn-glycerol (DG). Each isozyme of PKC is involved in diverse biological events, indicating that it serves as a novel therapeutic target. Since PKC isozymes contain two possible binding sites of tumor promoters and DG (C1A and C1B domains), the design of agents with binding selectivity for individual PKC C1 domains is a pressing need. We developed a synthetic C1 peptide library of all PKC isozymes for high-throughput screening of new ligands with such binding selectivity. This peptide library enabled us to determine that indolactam and benzolactam compounds bound to the C1B domains of novel PKC isozymes (delta, epsilon, eta, theta) in some selective manner, unlike phorbol esters and DG. Simpler in structure and higher in stability than the other potent tumor promoters, a number of indolactam and benzolactam derivatives have been synthesized to develop new PKC isozyme modulators by several groups. We focused on the amide function of these compounds because recent investigations revealed that both the amide hydrogen and carbonyl oxygen of indolactam-V (ILV) are involved in hydrogen bonding with the C1B domains of PKCdelta. Synthesis of several conformationally fixed analogues of ILV led to the conclusion that the trans-amide restricted analogues with a hydrophobic chain at an appropriate position (2,7) are promising leads with a high binding selectivity for novel PKC isozyme C1B domains. We also developed a new lactone analogue of benzolactam-V8 (17) which shows significant binding selectivity for the C1B domains of PKCepsilon and PKCeta. Furthermore, our synthetic approach with the PKC C1 homology domains clarified that diacylglycerol kinase beta and gamma are new targets of phorbol esters. PMID- 15134489 TI - Inhibition of anti-apoptotic Bcl-2 family proteins by natural polyphenols: new avenues for cancer chemoprevention and chemotherapy. AB - Amongst the most promising chemopreventive agents, certain natural polyphenols have recently received a great deal of attention from the scientific community, nutritionists, the pharmaceutical industry and the public, due to their demonstrated inhibitory activity against tumorigenesis. In view of their anticancer properties, these compounds also hold great promise as potential chemotherapeutic agents. However, to translate these chemopreventive agents into chemotherapeutic compounds, their exact mechanism of action must be delineated. The aim of this manuscript is to review recent findings suggesting that certain natural products bind and antagonize the anti-apoptotic effects of Bcl-2 family proteins such as Bcl-x(L) and Bcl-2. We will summarize recent studies that were aimed at the identification of the molecular targets of natural polyphenols and at the characterization of their mechanism of action on a molecular level. We will emphasize the importance of these findings that resides not only in the opportunity for the development of novel cancer treatments with these compounds, but also in the structural information that can be used for the design and development of novel and more effective semi-synthetic analogues. The finding reviewed here should encourage the study of possible direct effects of other dietary compounds on Bcl-2 family proteins. PMID- 15134490 TI - Anticancer agents: tumor cell growth inhibitory activity and binary QSAR analysis. AB - The tumor cell growth inhibitory activities (log 1/GI(50)) of 166 anticancer agents studied at the National Cancer Institute (NCI) in vitro anticancer screening program have allowed us to analyze the relative importance of physicochemical parameters in influencing the inhibitory activities. Increased molecular weight, as measured by the logarithm of molecular weight (log MW), is found to be an important contributor to the tumor cell growth inhibitory activities. The tumor cell growth inhibitory activities in different subpanels of the tumor cells are highly inter-correlated with each other. A simple binary quantitative structure-activity relationship (QSAR) model was derived from the 166 anticancer drugs, based on the tumor cell growth inhibitory activities transformed into a binary (active or inactive) data format. The model obtained can be tested with additional new data, and may be useful to identify active compounds from a large compound library to be included in high throughput screening. PMID- 15134491 TI - Chemistry and biology of curacin A. AB - Many natural and synthetic compounds bind to tubulin, an ubiquitous globular protein that provides the building blocks for the cellular microtubule network that controls chromosome segregation during mitosis, vesicle movements, intracellular transport of organelles, ciliar and flagellar movement, and maintenance of cell shape. Since the isolation of the antimitotic marine natural product curacin A in 1994, synthetic work on this colchicine-site binding agent has been intense, but only recently have synthetic derivatives been identified that match its potency for tubulin polymerization inhibition and its high level of growth inhibition in cancer cell lines. In addition to several total synthesis efforts, combinatorial libraries were constructed using solution phase and fluorous scavenging approaches. Low water-solubility and lack of chemical stability represent strong detriments for the clinical development of curacin A, but synthetic analogs with improved bioavailability might ultimately probe the paradigm for anticancer efficacy of colchicine-site tubulin binding agents. PMID- 15134501 TI - Synthetic retinoids and their nuclear receptors. AB - In addition to all-trans-retinoic acid and its 9 and 13-cis isomers, four synthetic retinoids are currently available to treat diseases of hyperproliferation, such as acne, psoriasis, and actinic keratosis, or cancers such as acute promelocytic leukemia, cutaneous T-cell lymphoma, and squamous or basal cell carcinoma. The retinoids extert their antiproliferative effects by interacting with their retinoic acid and retinoid X receptors that act as ligand inducible transcription factors. These homologous receptors function either directly on retinoid response elements or indirectly by modifying the responses of other transcription factors. Their major domains for binding DNA and their ligands have been characterized by either nuclear magnetic resonance spectroscopy or X-ray crystallography. The identification and design of synthetic retinoids are overviewed, as are their selective interactions with specific retinoid receptor subtypes and their clinical effects against cancer. Emphasis is placed on the retinoid X receptors and their ligands. PMID- 15134503 TI - New molecules and strategies in the field of anticancer agents. AB - The conventional chemotherapy is mostly based on the evidence that proliferating cells are more sensitive to anticancer agents than non-dividing cells. This is the main reason why these compounds are not tumour specific and their selectivity is generally in favour of rapidly growing cells (haematopoietic or intestine. i.e.) rather than discriminating against any fundamental biological difference between normal and tumour cells. The critical issue is at present to identify how tumour cells differ from normal cells and how those differences can be exploited therapeutically for designing and synthesising new drugs with a selective mechanism of action and thus with an improved therapeutic index. This topic and the strategies to identify these new targets will be discussed in details in the review. The expanding knowledge on molecular biology of cancer cells has allowed in the last years the identification of different molecular pathways altered in cancer that could be exploited as potential therapeutic targets. For most of the pathways previously disclosed it has been a problem to develop selective molecules with a relevant clinic impact. To target those specific genetics defects, different kind of molecules (antibodies, "antisense oligonucleotides", short peptides and small molecules) have been made and some of them are currently under investigation. This review will be focused mainly on three different classes of compounds: I. Compounds designed to hit or inhibit crucial molecular targets. II. Novel DNA minor groove binders. III. Products of marine origin that exhibit novel mode of action. PMID- 15134502 TI - Non-small cell lung cancer: from cytotoxic systemic chemotherapy to molecularly targeted therapy. AB - Surgery is the only method of cure in lung cancer. Seldom its application with radical intent is possible. Despite the efforts aimed at integrating all the therapeutic strategies, the overall outcome of the management of this disease remains disappointing. For this reason, in the last three decades, thousands of preclinical and clinical attempts have been realised in order to investigate any possible way to cure this disease and significant steps forward have been made on the basis of the increasing "molecular knowledge" in the so called "post-genomic era". Particularly the impressive step forward in the biological characterization of cancer as a result of genetic/epigenetic multistep process has brought in a multitude of variables with staggering classification potentialities. "Benchside" and "bedside" scientists have assembled in functional teams to move the common efforts "translationally" to bridge basic and clinical research for a mutual synergistic enhancement. This paper represents the effort of a lung cancer focused translational research team made up of molecular biologists, medical oncologists and thoracic surgeons to achieve a comprehensive, but simple, review of the current status of the shift from cytotoxic to molecularly targeted therapy in lung cancer treatment potentially useful in the planning of translational research trials. PMID- 15134504 TI - Soy saponins and the anticancer effects of soybeans and soy-based foods. AB - While the cancer protective effect of soy-based diets has been the subject of numerous studies, the constituents of soy that may give rise to this effect remain elusive. Recent publications describing anticancer activity of crude and purified soybean saponins have sparked a renewed interest in these compounds. In this review, I summarize the epidemiological studies concerning the cancer protective effects of soy and the efforts to elucidate the constituents responsible for this effect. The recent reports of the anticancer activity of soy saponins is placed in context with reports of promising anticancer activity of structurally related non-dietary saponins from other legumes. While recent studies have demonstrated a direct effect of soy saponins on cancer cells, alternative mechanisms of cancer prevention by these agents are also discussed. It is concluded that the soy saponins may represent promising leads both in terms of elucidating the soy constituents involved in the cancer protective effect of soy as well as in the discovery of anticancer agents with novel mechanisms of action. PMID- 15134505 TI - Quantitative structure-activity relationship study of histone deacetylase inhibitors. AB - Histone deacetylases (HDACs) play a critical role in gene transcription and have become a novel target for the discovery of drugs against cancer and other diseases. During the past several years there have been extensive efforts in the identification and optimization of histone deacetylase inhibitors (HDACIs) as novel anticancer drugs. Here we report a comprehensive quantitative structure activity relationship (QSAR) study of HDACIs in the hope of identifying the structural determinants for anticancer activity. We have identified, collected, and verified the structural and biological activity data for 124 compounds from various literature sources and performed an extensive QSAR study on this comprehensive data set by using various QSAR and classification methods. A highly predictive QSAR model with R(2) of 0.76 and leave-one-out cross-validated R(2) of 0.73 was obtained. The overall rate of cross-validated correct prediction of the classification model is around 92%. The QSAR and classification models provided direct guidance to our internal programs of identifying and optimizing HDAC inhibitors. Limitations of the models were also discussed. PMID- 15134506 TI - On the selectivity of 5-aminolevulinic acid-induced protoporphyrin IX formation. AB - Due to its capability to induce accumulation of protoporphyrin IX (PpIX) selectively in a multitude of different pathologies, 5-aminolevulinic acid (ALA) and its derivatives have attracted enormous attention in the field of photodynamic therapy (PDT) in the past two decades. The photochemical and photophysical properties of PpIX have been used for the fluorescence photodetection and photodynamic treatment of neoplasms in several medical indications in which conversion of ALA into PpIX seems to take place preferentially. Recently, this has led to the approval of this therapy for the treatment of actinic keratosis and basal cell carcinoma. When applied topically or systemically, ALA bypasses the negative feedback control that haem exerts on the enzyme ALA synthase (ALAS), which catalyses the natural production of this delta-amino acid, thereby temporarily boosting the generation of PpIX, the direct precursor of haem. Despite considerable interest in this treatment methodology, only little is known concerning the reasons for the selective accumulation of PpIX in neoplastic tissue upon ALA administration. Following an introduction into the biochemical as well as the chemical principles of haem synthesis, the present review tries to summarise experimental evidences of the mechanisms underlying preferential production of PpIX in neoplastic tissues. Thereby, morphological, environmental, enzymatic, as well as cell-specific factors will be discussed. PMID- 15134507 TI - Establishing biomarkers of oxidative stress: the measurement of hydrogen peroxide in human urine. AB - Hydrogen peroxide (H(2)O(2)) can be detected in freshly-voided human urine from healthy subjects and has been proposed as a "biomarker" of oxidative stress. This paper summarizes our studies to examine the extent to which urinary H(2)O(2) measurement fulfils the criteria for the "ideal biomarker". Levels of H(2)O(2), standardised for creatinine, varied widely between subjects. In most subjects, levels also varied considerably when measurements were made at different times and on different days. A reproducible increase in urinary H(2)O(2) was detected in all subjects examined after drinking coffee, a beverage rich in H(2)O(2). By contrast, green tea decreased urinary H(2)O(2) levels. We conclude that the H(2)O(2) in coffee is not excreted into urine. Instead, hydroxyhydroquinone from coffee is absorbed, excreted and oxidises in urine to produce H(2)O(2). No other confounders of urinary H(2)O(2) have been identified to date. Work is underway to compare H(2)O(2) levels with variations in other biomarkers of oxidative damage, to test the possibility that there are daily or other periodic variations in oxidative damage rates. PMID- 15134508 TI - Oxidative stress, antioxidants, and endothelial function. AB - Endothelial dysfunction, characterized by a loss in nitric oxide bioactivity, is an early event in the development of atherosclerosis and determines future vascular complications. Emerging evidence suggests a causal role for oxidative stress in this process. Reactive oxygen species can directly inactivate nitric oxide, modulate protein function and act as cellular signaling molecules. These events contribute to the initiation and progression of endothelial dysfunction. Considerable data also indicates that antioxidant compounds limit oxidative damage and restore endothelial function. The purpose of this review is to discuss these data and suggest novel approaches for lowering the oxidative stress in the vasculature. PMID- 15134509 TI - Role of oxidant species in aging. AB - Organisms are constantly exposed to many different forms of reactive oxygen species and reactive nitrogen species that damage proteins, nucleic acids, and lipids, leading to loss of biological function. The possibility that reactive oxygen/nitrogen-mediated protein damage contributes to the aging process is supported by results of many studies showing that aging is associated with the accumulation of such protein damage. Summarized here are results of studies, showing that the accumulation of,protein damage is a complex function of a multiplicity of factors that govern the intracellular levels of reactive oxygen/nitrogen species, on the one hand, and a multiplicity of factors that govern the degradation and/or repair of damaged proteins, on the other. Basic mechanisms involved in the modification of proteins by various forms of reactive oxygen/nitrogen species are also discussed. PMID- 15134510 TI - Non-antioxidant activities of vitamin E. AB - Molecules in biological systems often can perform more than one function. In particular, many molecules have the ability to chemically scavenge free radicals and thus act in the test tube as antioxidant, but their main biological function is by acting as hormones, ligands for transcription factors, modulators of enzymatic activities or as structural components. In fact, oxidation of these molecules may impair their biological function, and cellular defense systems exist which protect these molecules from oxidation. Vitamin E is present in plants in 8 different forms with more or less equal antioxidant potential (alpha , beta-, gamma-, delta-tocopherol/tocotrienols); nevertheless, in higher organisms only alpha-tocopherol is preferentially retained suggesting a specific mechanism for the uptake for this analogue. In the last 20 years, the route of tocopherol from the diet into the body has been clarified and the proteins involved in the uptake and selective retention of alpha-tocopherol discovered. Precise cellular functions of alpha-tocopherol that are independent of its antioxidant/radical scavenging ability have been characterized in recent years. At the posttranslational level, alpha-tocopherol inhibits protein kinase C, 5 lipoxygenase and phospholipase A2 and activates protein phosphatase 2A and diacylglycerol kinase. Some genes (e. g. scavenger receptors, alpha-TTP, alpha tropomyosin, matrix metalloproteinase-19 and collagenase) are modulated by alpha tocopherol at the transcriptional level. alpha-Tocopherol also inhibits cell proliferation, platelet aggregation and monocyte adhesion. These effects are unrelated to the antioxidant activity of vitamin E, and possibly reflect specific interactions of alpha-tocopherol with enzymes, structural proteins, lipids and transcription factors. Recently, several novel tocopherol binding proteins have been cloned, that may mediate the non-antioxidant signaling and cellular functions of vitamin E and its correct intracellular distribution. In the present review, it is suggested that the non-antioxidant activities of tocopherols represent the main biological reason for the selective retention of alpha tocopherol in the body, or vice versa, for the metabolic conversion and consequent elimination of the other tocopherols. PMID- 15134511 TI - Lipoic acid as a potential therapy for chronic diseases associated with oxidative stress. AB - alpha-Lipoic acid (LA), a naturally occurring dithiol compound, has long been known as an essential cofactor for mitochondrial bioenergetic enzymes. Aside from its enzymatic role, in vitro and in vivo studies suggest that LA also acts as a powerful micronutrient with diverse pharmacologic and antioxidant properties. Pharmacologically, LA improves glycemic control, polyneuropathies associated with diabetes mellitus, and effectively mitigates toxicities associated with heavy metal poisoning. As an antioxidant, LA directly terminates free radicals, chelates transition metal ions (e.g. iron and copper), increases cytosolic glutathione and vitamin C levels and prevents toxicities associated with their loss. These diverse actions suggest that LA acts by multiple mechanisms both physiologically and pharmacologically, many of which are only now being explored. Herein, we review the known biochemical properties of LA with particular reference to how LA may be an effective agent to ameliorate certain pathophysiologies of many chronic diseases. PMID- 15134512 TI - Potential therapeutic effect of antioxidant therapy in shock and inflammation. AB - Oxidative stress results from an oxidant/antioxidant imbalance, an excess of oxidants and/or a depletion of antioxidants. A considerable body of recent evidence suggests that oxidant stress plays a major role in several aspects of acute and chronic inflammation and is the subject of this review. Immunohistochemical and biochemical evidence demonstrate the significant role of reactive oxygen species (ROS) in acute and chronic inflammation. Initiation of lipid peroxidation, direct inhibition of mitochondrial respiratory chain enzymes, inactivation of glyceraldehyde-3-phosphate dehydrogenase, inhibition of membrane Na+/K+ ATP-ase activity, inactivation of membrane sodium channels, and other oxidative protein modifications contribute to the cytotoxic effect of ROS. All these toxicities are likely to play a role in the pathophysiology of shock, inflammation and ischemia and reperfusion. (2) Treatment with either peroxynitrite decomposition catalysts, which selectively inhibit peroxynitrite, or with SODm's, which selectively mimic the catalytic activity of the human superoxide dismutase (SOD) enzymes, have been shown to prevent in vivo the delayed tissue injury and the cellular energetic failure associated with inflammation. ROS (e.g., superoxide, peroxynitrite, hydroxyl radical and hydrogen peroxide) are all potential reactants capable of initiating DNA single strand breakage, with subsequent activation of the nuclear enzyme poly (ADP ribose) synthetase (PARS), leading to eventual severe energy depletion of the cells, and necrotic-type cell death. Antioxidant treatment inhibits the activation of PARS, and prevents the organ injury associated with acute and chronic inflammation. PMID- 15134513 TI - Oxidative stress and cell signalling. AB - An increasing body of evidence from animal models, human specimens and cell lines points to reactive oxygen species as likely involved in the pathways, which convey both extracellular and intracellular signals to the nucleus, under a variety of pathophysiological conditions. Indeed, reactive oxygen species (ROS), in a concentration compatible with that detectable in human pathophysiology, appear able to modulate a number of kinases and phosphatases, redox sensitive transcription factors and genes. This type of cell signalling consistently implies the additional involvement of other bioactive molecules that stem from ROS reaction with cell membrane lipids. The present review aims to comprehensively report on the most recent knowledge about the potential role of ROS and oxidised lipids in signal transduction processes in the major events of cell and tissue pathophysiology. Among the lipid oxidation products of ROS dependent reactivity, which appear as candidates for a signalling role, there are molecules generated by oxidation of cholesterol, polyunsaturated fatty acids and phospholipids, as well as lysophosphatidic acid and lysophospholipids, platelet activating factor-like lipids, isoprostanes, sphingolipids and ceramide. PMID- 15134514 TI - Peptidomimetics - antagonists of the fibrinogen receptors: molecular design, structures, properties and therapeutic applications. AB - The platelet aggregation is a crucial step in a pathophisiology of thromboses, leading to development of cardio-vascular diseases (myocardial infarction, transient ischemic attacks, strokes, etc.). The final step in the aggregation is the binding of fibrinogen to receptor - glycoprotein IIb/IIIa (GP IIb/IIIa) on the surface of activated platelets. In recent years the increasing attention is paid to the role of fibrinogen antagonists in the prevention of thrombosis. The search for these compounds is based on the molecular design of structures mimicking some fragment of RGD (Arg-Gly-Asp) sequence, responsible for the binding of fibrinogen to GP IIb/IIIa. Up to now, a large number of potent and selective GP IIb/IIIa antagonists, including non-peptide inhibitors are identified (derivatives of benzodiazepines, aminobenzamidinosuccinyles, isoxazolines, isoquinolines). The modification of natural peptide structures for obtaining of more active and selective fibrinogen receptor antagonists is realized in several ways: substitution of main pharmacophores of RGD sequence; cyclization of RGD-containing peptides; design of conformationally constrained peptidomimetics. For the treatment of chronic cardio-vascular diseases, the clinic needs high orally active RGD-peptidomimetics. This task is realized by obtaining of prodrugs on the base of the most potent RGD-mimetics. In our laboratory the molecular design and synthesis of non-peptide fibrinogen receptor antagonists were carried out. The series of RGD-mimetics on the basis of 4-oxo (piperazine-1-yl)butyric acid as Arg-mimetic and beta-aryl-beta-alanines as Asp Phe-mimetics were synthesized. Obtained RGD-mimetics showed a high antiaggregatory activity in vitro experiments with IC(50)values of 10(-7) - 10( 9) M. PMID- 15134515 TI - Recent developments in the chemistry of potassium channel activators: the cromakalim analogs. AB - Potassium channels play a crucial role in controlling the cell membrane potential. Among the different varieties of K(+) channels, the ATP-sensitive potassium channels (K(ATP) channels) have been characterized in numerous cell types, such as skeletal and smooth muscle cells, endocrine cells, cardiac cells and central neurons. Several molecules are known to activate K(ATP) channels and have been named "potassium channel openers" (PCOs). Such compounds may have a wide therapeutic potential and a few drugs are currently used as antihypertensive agents. Different chemical series of PCOs have been explored. This heterogeneous group of organic compounds comprises the benzopyran series including potent vasorelaxant drugs, such as cromakalim. The latter compound, a typical example of potassium channel opener, exerts its biological effect by activating K(ATP) channels. This review presents recent developments in the chemistry of cromakalim analoges and reports chemical aspects governing their potency and tissue selectivity. PMID- 15134516 TI - New developments on thromboxane and prostacyclin modulators part I: thromboxane modulators. AB - The pathogenesis of numerous cardiovascular, pulmonary, inflammatory, and thromboembolic diseases can be related to arachidonic acid (AA) metabolites. One of these bioactive metabolites of particular importance is thromboxane A(2) (TXA(2)). It is produced by the action of thromboxane synthase on the prostaglandin endoperoxide H(2)(PGH(2)), which results from the enzymatic degradation of AA by the cyclooxygenases. TXA(2) is a potent inducer of platelet aggregation, vasoconstriction and bronchoconstriction. It is involved in a series of major pathophysiological states such as asthma, myocardial ischemia, pulmonary hypertension, and thromboembolic disorders. Therefore, TXA(2) receptor antagonists, thromboxane synthase inhibitors and drugs combining both properties have been developed by several pharmaceutical companies since the early 1980s. Several compounds have been launched on the market and others are under clinical evaluation. Moreover, the recent literature reported the interest of thromboxane modulators, which combine another pharmacological activity such as, platelet activating factor antagonism, angiotensin II antagonism, or 5-lipoxygenase inhibition. In this review, we will propose a description of the recently described thromboxane modulators of major interest from both a pharmacological and a chemical point of view. PMID- 15134517 TI - New developments on thromboxane and prostacyclin modulators part II: prostacyclin modulators. AB - Prostacyclin (PGI(2)) is a potent endogenous inhibitor of platelet function and possesses a strong vasodilator effect. Furthermore, prostacyclin is currently presented as the physiologic antagonist of thromboxane A(2)(TXA(2)), which exhibits pro-aggregatory and vasoconstrictor properties. So, the balance between PGI(2) and TXA(2) production is crucial for the cardiovascular system. Indeed, an imbalance in the production or effect of these products is deleterious for the circulatory system and can lead to characterized vascular diseases such as hypertension, stroke, atherosclerosis or myocardial infarction. Although the biological effects of PGI(2) are considered to be clinically useful, its use as therapeutic agent is largely limited by both its chemical and metabolic instability. Actually, several prostacyclin agonists have been synthesized and pharmacologically evaluated. Among these, some have been clinically evaluated as therapeutic agents in several vascular diseases. This review focuses on the latest chemical and pharmacological developments in the field of the prostacyclin agonists. PMID- 15134518 TI - Biological activity and delivery of peptide nucleic acids (PNA)-DNA chimeras for transcription factor decoy (TFD) pharmacotherapy. AB - Peptide nucleic acids (PNAs) are recently described DNA mimics, in which the sugar-phosphate backbone is replaced by N-(2-aminoethyl)glycine units. These molecules efficiently hybridize with complementary DNA, forming Watson-Crick double helices. In addition, the interest of PNAs and PNA-based analogs is related to the fact that they are resistant to DNases and proteinases. While applications of PNAs as antisense and antigene molecules in non-viral gene therapy are well documented, their effects as potential transcription factor decoy (TFD) molecules is not demonstrated. PNA/PNA and PNA/DNA duplex are not suitable for TFD. In fact, PNA/PNA duplex does not recognize transcription factors, while, in the case of PNA/DNA hybrids containing nuclear factor binding sites, the interaction with transcription factors is unstable. By sharp contrast, double stranded molecules based on PNA-DNA chimeras exhibit strong TFD activity, display enzymatic stability in serum and cellular extracts and can be delivered to target cells after complexation with liposomes and microspheres. The TFD molecules based on PNA-DNA chimeras can be further engineered by addition of short peptides facilitating cell penetration and nuclear localization. Therefore, these engineered molecules could be of great interest for in vivo experiments on non-viral gene therapy of a variety of diseases, including neoplastic and viral diseases, for which the TFD approach has been already demonstrated as a very useful strategy. PMID- 15134519 TI - Recent advances in artemisinin and its derivatives as antimalarial and antitumor agents. AB - Artemisinin, the first and last naturally occurring 1, 2, 4-trioxane originated from Artemisia annua, L. and its derivatives are a potent class of antimalarial drugs. The clinical efficacy of these drugs is characterized by an almost immediate onset and rapid reduction of parasitemia, and it is high in such areas as well where multidrug-resistance is rampant. Furthermore, artemisinin and many of its analog possess not only antiparasitic effect against Plasmodium falciparum, Schistosoma japonicum and Clonorchis sinensi but also immuno modulation effects, and antitumor activities. This review covers the chemistry of artemisinin including synthesis of acetal-, non acetal-type C-12 analogs, C-11- and C-13 derivatives from artemisitene, ring-contracted derivatives, dimers, and trimers. Modes of biological action of artemisinin - derived analogs are also reviewed. The main objective of this article is to review the literatures of recent progress taken place in chemistry, mode of biological actions of artemisinin, and its derivatives as antimalarial and antitumor agents during the last three years (1999-2001). PMID- 15134520 TI - Semicarbazide-sensitive amine oxidase: current status and perspectives. AB - Semicarbazide-sensitive amine-oxidase (SSAO) is present in various human tissues and in plasma. Oxidative deamination of short-chain aliphatic amines is catalyzed by this enzyme to afford the corresponding aldehydes, ammonia and hydrogen peroxide. Methylamine and aminoacetone have been recognized to be physiological substrates for SSAO. There are several pathological states where increased serum SSAO activity have been found, such as diabetes mellitus, congestive heart failure, multiple types of cerebral infarction, uraemia, and hepatic cirrhosis. The role of SSAO in pathophysiology of diabetes has been most extensively investigated. The elevated formation of the potentially cytotoxic products of the enzyme may contribute to the endothelial injury of blood vessels, resulting in the early development of severe atherosclerosis; it may also contribute to the pathogenesis of diabetic angiopathy. It is now suggested that SSAO inhibitors may prevent the development of atherosclerosis and diabetic complications as well. Inhibitors can be conveniently subdivided into the main groups of hydrazine derivatives, arylalkylamines, propenyl- and propargylamines, oxazolidinones, and haloalkylamines. Of them, aryl(alkyl)hydrazines, and 3-halo-2-phenylallylamines are generally very strong SSAO inhibitors. Most of these inhibitors of SSAO have been originally developed for other purposes, or they are simple chemical reagents with highly reactive structural element(s); these compounds have not been able to fulfil all criteria of high potency, selectivity, and acceptable toxicity. New potent compounds with selectivity and low toxicity are needed, which may prove useful tools for understanding the roles and function of SSAO, or they may even be valuable substances for treatment of various diseases. PMID- 15134521 TI - Mitochondrial superoxide dismutase: a promising target for new anticancer therapies. AB - Compelling experimental and epidemiological evidence involves oxygen radicals in carcinogenesis, acting reactive oxygen species both as endogenous genotoxins during cell initiation and as messenger molecules in mitogenesis and in tumor promotion. Moreover, oxidants stimulate neoangiogenesis, which is a prerequisite for tumor growth. However, while several natural as well as synthetic antioxidant compounds appear to be chemopreventive in mutagenicity assays, antioxidant-based treatments for the prevention or cure of cancer have led to non-conclusive if not disappointing results. This is likely due to the fact that oxygen radicals have also a major role in the natural defences against the propagation of cancer cells, i.e. tumor cell apoptosis and immune surveillance, and mediate the beneficial cytotoxic effect of both the chemo-and radio-therapy. In recent years, the mitochondrial antioxidant enzyme, Manganous Superoxide Dismutase (MnSOD), has received a growing attention as a negative modulator of cellular apoptosis and as a survival factor for cancer cells. In fact, while overexpression of this enzyme in cancer cells decreases proliferation and tumor incidence in transgenic models, it is clear that even small amounts of this enzyme are crucial for cell resistance to inflammatory stimuli and anticancer drugs, and prevent oncogene induced apoptosis triggered by the tumor suppressor protein p53. A previously unexpected oncogenic potential of MnSOD is also suggested by the elevated levels of this enzyme in several classes of human neoplasms, in a fashion which often correlates with the degree of their malignancy. This review focuses on the debated issue of the pro- and/or anti-tumoral effect of MnSOD, with special emphasis on recent observations suggesting that pharmacological inhibition of MnSOD may represent an effective strategy to selectively kill cancer cells and to circumvent their resistance to the commonly used anticancer treatments. PMID- 15134522 TI - Chemistry and biology of cyclic depsipeptides of medicinal and biological interest. AB - Cyclodepsipeptides comprise a wide variety of cyclic peptides of natural origin and are characterized by the occurrence of at least one ester linkage. The great interest that this class of natural products has elicited in scientific community is explained by their wide range of biological activities, intriguing mechanisms of action and attractive molecular architecture. For example, they display a variety of biological effects, such as immunosuppressant, antibiotic, antifungi, antiinflammatory or antitumoral activities. In addition, many of these cyclic depsipeptides represent useful tools for the research of biological processes involved in cellular regulation. The present review deals with the most interesting aspects of the biology and the chemistry of some of these compounds. PMID- 15134523 TI - The relation between the chemical structure of flavonoids and their estrogen-like activities. AB - Estrogens are steroid hormones exhibiting a broad range of physiological activities, which are important in the homeostatic regulation of many cellular and biochemical events. Estradiol (E(2)) is an estrogen which is active in the development of the mammary glands and the uterus, in the maintenance of pregnancy and bone density, in protection from cardiovascular diseases, and in the relief of menopausal symptoms. However, it can also stimulate malignant growths, and hence contribute to the development of estrogen-dependent tumors, such as cancer of the breast and uterus. Phytoestrogens are plant-derived chemicals with estrogen-like activities, which could have a beneficial role in humans against estrogen deficiency. Several studies relate the lower incidence of estrogen deficiency-related disease among women in the Eastern word to a diet rich in phytoestrogens. This compound comprises a variety of structurally diverse chemicals, with flavonoids as their largest group. In the present review we summarize knowledge gained on the relationship between the structure of flavonoids to their ability to mimic estrogen activity, together with the methods adapted to perform these studies. The methods reviewed are qualitative and quantitative measurements of the estrogen-like activities of phytoestrogens, superposition analysis, docking analysis and quantitative structure-activity relationship (QSAR) models. The potential of various new flavonoid derivatives in hormone replacment therapy is discussed. PMID- 15134525 TI - Clinical impact of gene expression profiling on oncology diagnosis, prognosis, and treatment. AB - Genomics has enabled the examination of the totality of disease at the transcriptome level. Dependent upon a myriad of genetic aberrations for its pathogenesis, cancer has been the focus of gene expression profiling studies that have highlighted the potential clinical applications of this technology. This type of molecular profiling has the potential to enhance the ability of pathologists and oncologists to correctly classify tumors, not just into existing subgroups which may or may not have clear prognostic implications, but into new groups which carry predictable correlations with outcomes. Ultimately, these outcome predictions can be tied to specific treatment regimens, allowing clinicians to predict at the time of diagnosis to which therapy a given patient may best respond. Although this ultimate goal of personalized therapy remains in the future, the numerous studies to date have clearly demonstrated the overall feasibility of this approach. This review will showcase a few of these studies in several key tumor types with the goal of demonstrating which type of studies have been conducted and what types of results are currently possible. PMID- 15134524 TI - Proanthocyanidins in health care: current and new trends. AB - Polyphenolic compounds are widely distributed in higher plants and are an integral part of the human diet. Recent interest in these substances has been stimulated by their potential health benefits, which are believed to arise mainly from their antioxidant activity. In the past years, the antioxidant activity of flavonoids has been studied in detail. An important but often overlooked group of polyphenols is that of the proanthocyanidins. Therefore, the present review is focused mainly on the antioxidant activity of proanthocyanidins and its relevancy in vivo. The three most important mechanisms of their antioxidant action will be discussed, i.e. free radical scavenging activity, chelation of transition metals, and inhibition of enzymes. In addition, the protective role of proanthocyanidins against lipid peroxidation and peroxynitrite, as well as their antimicrobial properties will be discussed. To study the in vivo relevancy of the proanthocyanidin activities, the knowledge of their pharmacokinetic parameters is crucial. Although bioavailability and metabolism data on polyphenols in general and proanthocyanidins in particular are still largely unavailable, the first reports indicate that at least monomers and smaller oligomeric procyanidins are absorbed. There is also considerable scientific and public interest in the important role that antioxidants may play in health care, e.g. by acting as cancer chemopreventive and anti-inflammatory agents and by reducing risk of cardiovascular mortality. Each of these aspects will be discussed, with special attention to the role of proanthocyanidins on apoptosis, gene expression and transcription factors, such as NF-kappa B. PMID- 15134526 TI - DNA microarrays as a tool in toxicogenomics. AB - Toxicogenomics is an emerging technology that defines the use of novel genomic techniques to investigate the adverse effects of xenobiotic on gene expression. Toxicogenomics is based on the fact that most of relevant toxicological effects of a compound affect directly or indirectly the gene expression. The most common methods to profile gene expression at the transcript level are Northern Blotting and the real-time PCR. While commonly used and well accepted, these techniques are now superseded by new technologies allowing the analysis of the expression for multiple genes simultaneously. DNA microarrays are now developed for simultaneous gene analysis but inherent to such multiple assays, their quantitative aspect and their relevance for toxicogenomics have been questioned. We will review here recent studies on their use for toxicogenomics and examine the possible future of such technology in complementation with the other toxicology methods. PMID- 15134527 TI - Strategies for immobilization of biomolecules in a microarray. AB - Recent advances in the generation of peptide and protein microarrays are reviewed, with special focuses on different strategies available for site specific immobilization of proteins and peptides. PMID- 15134528 TI - Protein microarrays and multiplexed sandwich immunoassays: what beats the beads? AB - Protein microarray technology allows the simultaneous determination of a large variety of parameters from a minute amount of sample within a single experiment. Assay systems based on this technology are currently applied for the identification, quantitation and functional analysis of proteins. Protein microarray technology is of major interest for proteomic research in basic and applied biology as well as for diagnostic applications. Miniaturized and parallelized assay systems have reached adequate sensitivity and hence have the potential to replace singleplex analysis systems. However, robustness and automation needs to be demonstrated before this technology will finally prove suitable for high-throughput applications. Miniaturized and parallelized sandwich immunoassays are the most advanced assays formats among the different protein microarray applications. Multiplexed sandwich immunoassays can be used for the identification of biomarkers and the validation of potential target molecules. In this review an overview will be given on the current stage of protein microarray technology with a special focus on miniaturized multiplexed sandwich immunoassays. PMID- 15134529 TI - Making sense of molecular signatures in the immune system. AB - The development of Functional Genomics technologies has opened new avenues to investigate the complexity of the immune system. Microarray technology has been particularly successful because of its relatively low cost and high genome coverage. Consequently to our ability to monitor the expression of a significant proportion of an organism genome, our understanding of the molecular dynamics behind cell differentiation and cell response has greatly improved. Molecular signatures associated to immune cells have provided important tools to investigate the molecular basis of diseases and have been often associated to diagnostic and prognostic markers. The availability of such large collection of data has stimulated the application of complex machine learning techniques in the attempt to link molecular signatures and cell physiology. Here we review the most recent developments in the analysis of molecular signatures in the immune system. PMID- 15134530 TI - Anti-endotoxin agents. 1. Development of a fluorescent probe displacement method optimized for the rapid identification of lipopolysaccharide-binding agents. 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 have shown using animal models of sepsis that the sequestration of circulatory LPS by small molecules is a therapeutically viable strategy. Assays reported previously in the literature do not lend themselves well to the rapid screening of large numbers of structurally diverse compounds. In this report, we describe a highly sensitive and robust fluorescent displacement assay using BODIPY TR cadaverine (BC), which binds specifically to the toxic center of LPS, lipid A, and is competitively displaced by compounds displaying an affinity for lipid A. The assay clearly discriminates subtle differences in the binding of polymyxin B, and its nonapeptide derivative, with LPS. The spectral properties of the BODIPY fluorophore are ideally suited for screening diverse structural classes of compounds, including those with conjugated aromatic groups, or with chromophores in the 260-500 nm range. The fluorescent probe: LPS complex is stable under physiologically relevant salt concentrations, resulting in the rapid rejection of spurious binders interacting via non-specific electrostatic interactions, and, therefore, in greatly improved dispersion of ED(50)values. PMID- 15134531 TI - Combinatorial synthesis of biheterocyclic benzimidazoles by microwave irradiation. AB - Liquid phasel synthesis of biheterocyclic benzimidazoles by controlled microwave irradiation was investigated. Polymer immobilized o-phenylenediamines was synthesized under microwave irradiation. The resulting PEG bound diamines was N acylated with 4-fluoro-3-nitrobenzoic acid selectively in primary aromatic amino moiety. Nucleophilic aromatic substitution of amide was performed with various amines then cyclized to form the first benzimidazole scaffold in acidic condition. Successive reduction, cyclization with isothiocyanates yielded 5 (benzimidazol-2-yl)benzimidazoles. The desired products were released from the polymer support to afford the tri-substituted bis-benzimidazoles in good yields and purity. PMID- 15134532 TI - The Akt pathway: molecular targets for anti-cancer drug development. AB - The serine/threonine kinase Akt functions intracellularly as a cardinal nodal point for a constellation of converging upstream signaling pathways, which involve stimulation of receptor tyrosine kinases such as IGF-1R, HER2/Neu, VEGF R, PDGF-R), and an assembly of membrane-localized complexes of receptor-PI-3K and activation of Akt through the second messenger PIP(3). The integration of these intracellular signals at the level of Akt and its kinase activity, regulates the phosphorylation of its several downstream effectors, such as NF-kappa B, mTOR, Forkhead, Bad, GSK-3 and MDM-2. These phosphorylation events in turn mediate the effects of Akt on cell growth, proliferation, protection from pro-apoptotic stimuli, and stimulation of neo-angiogenesis. Because Akt and its upstream regulators are deregulated in a wide range of solid tumors and hematologic malignancies, and in view of the aforementioned biologic sequelae of this pathway, the Akt pathway is considered a key determinant of biologic aggressiveness of these tumors, and a major potential target for novel anti cancer therapies. This review focuses on ongoing translational efforts to therapeutically target Akt and its biologic sequelae, either at the level of Akt itself or at the levels of its upstream regulators and downstream effectors. Because Akt is also important for proliferative and anti-apoptotic signaling pathways critical for normal cells, particular emphasis is placed on the fine tuning the targeting of individual components of this pathway to maximize the therapeutic index of anti-cancer strategies based on the PI-3K/Akt pathway. PMID- 15134533 TI - The role of cytochrome P450 in cytotoxic bioactivation: future therapeutic directions. AB - The cytochrome P450s are an essential group of enzymes involved in metabolism of drugs, foreign chemicals, arachidonic acid, cholesterol, steroids and other important lipids. The cytochrome P450 enzyme system is responsible for much of the phase I metabolism of chemotherapeutic agents. At the simplest level the detoxification properties of the cytochrome P450s are used to help clear a cytotoxic before it results in serious irreversible toxicity to the patient while at other levels the cytochrome P450s are involved to varying extents in drug bioactivation. This metabolism primarily occurs in organs and tissues of the body known to express cytochrome P450 ubiquitously (i.e. liver and gastrointestinal tract), but there is also evidence to suggest that it occurs within the tumor microenvironment due to localized, tumor specific expression of certain P450 isoforms. Several of today's currently prescribed cytotoxics (e.g. cyclophosphamide and tamoxifen) undergo systematic bioactivation by cytochrome P450, which often results in toxicity to the patient. The realization that many tumors have differential cytochrome P450 expression when compared to the corresponding normal tissue has allowed the rational design of the next generation of cytotoxic around cytochrome P450 enzymology. Several new agents now entering clinical trials (e.g. Phortress and AQ4N) are specifically designed to exploit tumor cytochrome P450, resulting in local bioactivation of the cytotoxic at the tumor site. Specific activation of pro-drugs by isoforms whose expression or particular catalytic activity is limited to cancer cells offers the possibility of truly targeted chemotherapy with minimized systemic toxicity. PMID- 15134534 TI - Leukotriene A4 hydrolase as a target for cancer prevention and therapy. AB - Leukotriene A4 hydrolase (LTA4H) is a bifunctional zinc enzyme with the activities of epoxide hydrolase and aminopeptidase. As an epoxide hydrolase, LTA4H catalyzes the hydrolysis of the epoxide LTA4 to the diol, leukotriene B4 (LTB4), which mainly functions as a chemoattractant and an activator of inflammatory cells. As an aminopeptidase, LTA4H may process peptides related to inflammation and host defense. In a chronic inflammation-associated animal model of esophageal adenocarcinoma, we have shown that LTA4H was overexpressed in tumor as compared to normal tissues. Bestatin, an LTA4H inhibitor, suppresses tumorigenesis in this animal model. Since LTA4H has long been regarded as an anti inflammatory target, we propose LTA4H as a target for prevention and therapy of cancers, especially those associated with chronic inflammation. Here we review the gene structure, expression, regulation and functions of LTA4H, as well as its involvement in carcinogenesis. We believe LTA4H/LTB4 may play an important role in chronic inflammation associated carcinogenesis by at least two mechanisms: a) the inflammation-augmenting effect on inflammatory cells through positive feedback mediated by its receptors and downstream signaling molecules; and b) the autocrine growth-stimulatory effect of LTB4 produced by epithelial cells, and the paracrine growth-stimulatory effect of LTB4 produced by inflammatory cells, on precancerous and cancer cells. Based on our present knowledge, inhibitors of LTA4H or antagonists of LTB4 receptors may be used alone or in combination with other agents (e.g., cyclooxygenase 2 inhibitors) in cancer prevention and treatment trials to test their effectiveness. PMID- 15134535 TI - Retinoids in cancer chemoprevention. AB - We review the therapeutic and preventive applications of a retinoid analog (vitamin A and its derivatives) for human cancers. Chemoprevention of cancer is an intervention in the carcinogenic process by chemical agents that block or reverse the malignant transformation of cells. Retinoids are prime candidates for cancer chemoprevention since cancer is characterized by abnormal growth with a lack of differentiation, which could be modified by retinoids. Retinoids exert their biological functions through nuclear receptors, retinoic acid receptor (RAR) and retinoid X receptor (RXR). A number of experimental and clinical studies have been performed in the past two decades with retinoids showing that they inhibit or reverse the carcinogenic process in some organs, including hematological malignancy as well as premalignant and malignant lesions in the oral cavity, head and neck, breast, skin and liver. We particularly focus upon the therapeutic application of all-trans RA (atRA) to acute promyelocytic leukemia (APL) and on the preventive approach to hepatocellular carcinoma (HCC) by a synthetic retinoid analog, acyclic retinoid. In both malignancies, malfunction of retinoid nuclear receptors is closely related to their carcinogenic process. In APL, a chromosomal translocation produces a chimeric protein between RAR alpha and a protein called promyelocyte leukemia protein (PML). PML-RAR alpha works as a dominant negative receptor in the leukemic cells, interfering with the normal function of RAR alpha and/or PML, which in turn results in the arrest of cell maturation at the stage of promyelocytes. Oral administration of atRA induces differentiation of promyelocytic leukemic cells to mature neutrophils, and leads to a high rates (over 90%) of complete remission. AtRA therapy has become standard in the treatment of APL. In the case of HCC, post-translational modification of RXR by phosphorylation impairs its function, which leads to uncontrolled cell growth. Acyclic retinoid suppresses the phosphorylation of RXR alpha, restores its function in the presence of its endogenous ligand, 9-cis RA, and thereby induces apoptosis of the cancer cells. Acyclic retinoid given orally successfully suppresses the development of second primary tumors in cirrhotic patients who undergo curative removal of preceding HCC. Eradication of (pre)malignant clones ('clonal deletion') from the liver is suggested as a mechanism of the chemopreventive effect. Further development of more effective retinoids as well as their use in combination with other classes of anticancer agents including immunopreventive drugs like interferons may provide strategies for cancer prevention. PMID- 15134536 TI - Targeting translation for treatment of cancer--a novel role for IRES? AB - Protein synthesis plays an important role in the regulation of cell proliferation. While the role of cap-dependent translation in cell transformation has been studied extensively another translation initiation mechanism, internal initiation of cellular mRNAs, emerged recently and is relatively unappreciated and poorly understood. Internal initiation is mediated by IRES elements that are found in the 5' untranslated region (5' UTR) of mRNA. Curiously, several oncogenes, growth factors and proteins involved in the regulation of programmed cell death contain IRES elements in their 5' UTRs. Internal initiation escapes many control mechanisms that regulate cap-dependent translation. In this review I will discuss the data supporting the hypothesis that selective translation of these factors may contribute to the survival of cancer cells under stressful situations, such as lack of nutrients, hypoxia, or therapy-induced DNA damage and contributes to the development and progression of cancer and to the establishment of cancer cells that are resistant to conventional therapies. PMID- 15134537 TI - The role of thymidine kinases in the activation of pyrimidine nucleoside analogues. AB - Deoxynucleoside analogues need activation by deoxynucleoside kinases to serve as antiviral or anticancer agents. Here we review the properties of cellular cytoplasmic thymidine kinase 1, mitochondrial thymidine kinase 2, the multisubstrate deoxynucleoside kinase from Drosophila melanogaster and Herpes virus 1 thymidine kinase. Important substrate activity relationships will be discussed. PMID- 15134538 TI - Structural requirements for efficient phosphorylation of nucleotide analogs by human thymidylate kinase. AB - Successive phosphorylation of nucleoside analog prodrugs to their triphosphate forms is required for the pharmacological activity of these compounds in the chemotherapeutic treatment of viral infections and cancer. Human thymidylate kinase (TMPK), apart from its essential physiological role in the biosynthesis of TTP, is also required for the activation of thymidine analogs, such as the clinically used anti-HIV prodrugs AZT and d4T. This enzyme is rate determining in the three-step cascade of AZT phosphorylation. Our structural work on human, yeast and E. coli TMPKs, in conjunction with sequence homology analyses and biochemical data, has demonstrated that three loops are crucial for the function of this enzyme: the first is the highly conserved P-loop motif, which binds and positions the phosphoryl groups of ATP, the second critical loop contains the DR(Y/H) motif that supplies a catalytic arginine and is also important for the binding and positioning of the magnesium ion complexed to ATP, and the third loop is the so-called Lid-region that is a flexible stretch which closes on ATP when it binds. Modifications of the sugar moieties of nucleoside monophosphates are shown to exert drastic effects on the enzyme's conformation and, thus, reduced activity. Our structural work on several TMPKs has formed the basis for generating mutants of human TMPK that are about 100 times more efficient in phosphorylating AZTMP. These enzyme variants could potentially be introduced into HIV-targeted cells in order to significantly improve AZT's antiviral activity. PMID- 15134539 TI - Antiviral nucleoside analogs phosphorylation by nucleoside diphosphate kinase. AB - The reaction of NDP kinase was studied in vitro with several antiviral derivatives, using kinetic steady state and presteady state analysis. The enzyme is highly efficient with natural nucleotides but most of the analogs are slow substrates. The catalytic efficiency, also related to the affinity of the analog, is mainly dependent on the presence of a 3'-OH group on the ribose moiety. PMID- 15134540 TI - Aryloxy phosphoramidate triesters as pro-tides. AB - We herein describe the development of aryloxy phosphoramidate triesters as an effective pro-tide motif for the intracellular delivery of charged bio-active antiviral nucleoside monophosphates. The review covers the discovery of such aryl phosphoramidates, their mechanism of action and structure-activity relationships. The application of this strategy to a range of antiviral nucleosides is highlighted. PMID- 15134541 TI - "Lock-in"-cycloSal-pronucleotides - a new generation of chemical Trojan Horses? AB - The cycloSal-concept is one example of a successful nucleotide delivering system (pronucleotide). For several nucleoside analogues, the cycloSal-approach improved the antiviral potency and the applicability of the nucleosides could be broadened. Here, a conceptional extension of the original design of the cycloSal system will be discussed. PMID- 15134542 TI - SATE pronucleotide approaches: an overview. AB - This review depicts in vitro and in vivo results obtained with nucleotide prodrugs (pronucleotides) bearing S-acyl-2-thioethyl (SATE) groups as esterase labile phosphate protections. New developments are illustrated by the design of mononucleoside mixed phosphoester derivatives leading to the selective intracellular delivery of the corresponding 5'-mononucleotide through two different enzyme-mediated activation steps. PMID- 15134543 TI - Designing a pronucleotide stratagem: lessons from amino acid phosphoramidates of anticancer and antiviral pyrimidines. AB - Phosphoramidate pronucleotides have proven to be an effective strategy for the intracellular delivery of nucleoside 5'-monophosphates. This review will summarize our efforts to understand the in vitro and in vivo behavior of phosphoramidate monoesters of 3'-azido-3'-deoxythymidine (AZT), 3'-fluoro-3' deoxythymidine (FLT) and 5-fluoro-2'-deoxyuridine (FUdR). Insights drawn from these studies have proved valuable for the future design of phosphoramidate-based pronucleotides. PMID- 15134544 TI - Discovery and validation of a new family of antioxidants: the aminopyrazine derivatives. AB - Coelenteramine (2-amino-1,4-pyrazine derivative), one of the metabolites of the oxidative degradation of coelenterazine (imidazolopyrazinone derivative), is endowed with excellent antioxidative properties towards ROS/RNS, like its mother compound. This crucial discovery, made during the study of natural bioluminescent compounds (luciferins), has stimulated the development of synthetic aminopyrazine derivatives as new leads in medicinal chemistry in the field of antioxidant-based therapies. Synthetic approaches, theoretical evaluation, radical scavenging properties in acellular and cellular tests, and in vivo evaluation are described, and illustrated with representative aminopyrazines. Tested compounds were inhibitors of lipid peroxidation and good quenchers of peroxynitrite. They efficiently protect isolated LDL against radical-induced damages. They prevent cell constituents (membranes, DNA) against injuries by various oxidative stressors (UV irradiation, hydroperoxide treatment, oxidized LDL toxicity). Lastly, aminopyrazines are remarkably active in the "hamster cheek pouch" assay (in vivo protection against ischemia-reperfusion damages). PMID- 15134545 TI - Bioactive carbohydrates and recently discovered analogues as chemotherapeutics. AB - Infectious diseases such as tuberculosis, malaria, the "simple flu" or HIV and HBV, are killing more than 50,000 people a day according to estimations by the World Health Organisation (WHO). Consequently, the development of biologically active agents in general, such as antibiotics and chemotherapeutics, is of great importance. Hand in hand with the understanding of the mechanisms of biological agents, structures carrying sugar moieties have become increasingly important during the last decades. This review will cover the most recent developments in the field of new antibiotics and synthetic agents containing carbohydrates which are active against tuberculosis and malaria. In addition, compounds having antiviral, antibacterial and anticancer properties will be examined. Compounds such as aminoglycosides, iminosugars, carbacycles, nucleosides, and other selected substance classes will be considered. PMID- 15134546 TI - Inhibitors of HIV-1 gene expression and transcription. AB - Human immunodeficiency virus type 1 (HIV-1) gene expression and transcription is an essential step in the viral life cycle, which is considered to be a possible target for inhibition of HIV-1 replication. Among the factors involved in this step, the cellular transcription factor nuclear factor (NF)-kappaB is the most potent inducer of HIV-1 gene expression, while the viral transactivator protein Tat seems to play a central role in sustaining a high level of HIV-1 replication. Another important mechanism of HIV-1 gene expression is the nuclear export control of viral mRNA conducted by the viral regulatory protein Rev. Various attempts have been undertaken to discover selective inhibitors of HIV-1 gene expression and transcription. Several small-molecule compounds were reported to inhibit Tat functions though blocking either the Tat/TAR RNA interaction or the kinase activity of cellular cofactors, such as cyclin T1/CDK9. In the case of Rev inhibitors, it appears to be more difficult to find them than Tat inhibitors, and only a few compounds have been identified as Rev inhibitors. However, the selectivity of these Tat and Rev inhibitors was not high enough to eliminate the cytotoxicity to the host cells. Since the signal transduction pathways leading to NF-kappaB activation are redox regulated, several antioxidants have been shown to block HIV-1 transcription. Although some of them have progressed into clinical trials in HIV-1-infected patients, the results were not conclusive. In addition, various compounds have been identified as inhibitors of HIV-1 gene expression and transcription, yet their precise mechanisms are still unknown. PMID- 15134547 TI - HIV co-receptors as targets for antiviral therapy. AB - The chemokine receptors CXCR4 and CCR5 are used as the main co-receptors by the T cell-tropic (CXCR4-dependent, X4) and macrophage-tropic (CCR5-dependent, R5) HIV 1 strains, respectively, for entering their target cells. The natural ligands for CXCR4, the CXC-chemokine SDF-1 and CCR5, the CC-chemokines RANTES, MIP-1alpha and MIP-1beta are described to inhibit viral entry. In this review we focus on chemokine receptor/HIV co-receptor inhibitors. Modified chemokines such as Met RANTES and AOP-RANTES showed antiviral activity against R5 viruses. Several low molecular weight CCR5 antagonists have been described (such as TAK-779 and SCH-C) with potent antiviral activity. The latter compound is also orally available and is able to decrease R5 viral load levels in HIV-infected subjects. Several peptidic compounds, such as T22 (an 18-mer), T134 (a 14-mer), ALX40-4C (a 9-mer) and CGP 64222 (also a 9-mer) have anti-HIV activity and have been identified as CXCR4 antagonists. Also, the HIV-1 Tat protein has been described as a "natural" CXCR4 antagonist with anti-HIV-1 activity. The most potent and specific CXCR4 antagonists are the bicyclam derivatives, which also potently inhibit X4 HIV replication. AMD3100 has proved to be a highly specific CXCR4 antagonist, which consistently blocks X4 viral replication in any target cell-type evaluated so far. AMD3100 was selected as the clinical drug candidate, which, after initial phase I (safety) studies, had proceeded to phase II (efficacy) trials. The compound dose-dependently inhibited X4 viruses after 10 days of continuous infusion of the drug. Recently, the orally bioavailable CXCR4 antagonist, AMD070, is presented as a candidate HIV drug. We believe that chemokine receptor antagonists will become important new antiviral drugs to combat AIDS. Both (CXCR4 and CCR5) chemokine receptor inhibitors will be needed in combination to inhibit viral replication and even in combinations of antiviral drugs that also target other aspects of the HIV replication cycle, such as reverse transcriptase and protease, to obtain optimum therapeutic effects. PMID- 15134548 TI - Nucleoside inhibitors of human immunodeficiency virus type 1 reverse transcriptase. AB - The development of novel compounds that can effectively inhibit both wild type and the most consensus resistant strains of human immunodeficiency virus type 1 (HIV-1) is the primary focus in HIV disease management. Combination therapy, comprising at least three classes of drugs, has become the standard of care for acquired immunodeficiency syndrome (AIDS) or HIV-infected individuals. The drug cocktail can comprise all three classes of HIV inhibitors, including nucleoside reverse transcriptase inhibitors (NRTI), non-nucleoside reverse transcriptase inhibitors (NNRTI) and protease inhibitors (PI). Due to their competitive mode of inhibition and requirement for metabolic activation, almost all NRTI drugs lack the virological potency of NNRTI or PI drugs. However, data from clinical trials indicate that sustained viral suppression could not be achieved with NRTI, NNRTI or PIs alone. Therefore, the NRTIs will remain essential components of highly active antiretroviral therapy (HAART) for the foreseeable future, because they enhance the virological potency of the regimen, they do not bind excessively to protein and most regimens are small pills/tablets given once a day. It has become apparent in recent years that the prolonged use of certain NRTIs exhibits adverse events as a class, limiting the length of time for which they can be safely used. Of major clinical concern is their association with the potentially fatal lactic acidaemia and hepatic steatosis. These class events, as well as individual drug effects, such as peripheral neuropathy, are linked to delayed mitochondrial destruction. In addition to toxicity, the development of resistance-conferring mutations against exposure to nucleoside analogs currently in use influences long term therapeutic benefits. Of critical importance for the evaluation of new NRTIs are recent studies showing that the efficiency of discrimination or excision by pyrophosphorolysis in the presence of nucleotides of a given NRTI is a key determinant in the emergence of one or the other resistance pathway. PMID- 15134549 TI - Current status of the non-nucleoside reverse transcriptase inhibitors of human immunodeficiency virus type 1. AB - Almost fifteen years ago, the first non-nucleoside reverse transcriptase (RT) inhibitor (NNRTI) lead compounds have been discovered. Nowadays, three NNRTIs are approved for treatment of HIV-1-infected individuals and several others are subject of (advanced) clinical trials. Although the NNRTIs target HIV-1 RT, they are clearly different from the nucleoside RT inhibitors (NRTIs). They are highly selective for HIV-1 and do not inhibit HIV-2 or any other (retro)virus. They target HIV-1 RT by a direct interaction without the need to be metabolised by cellular enzymes, and they interact at a site on the HIV-1 RT that is near to, but distant from, the substrate-binding site. The majority of NNRTIs share common conformational properties and structural features that let them fit in a hydrophobic pocket at the HIV-1 RT, which is nowadays well-characterized. A wide variety of crystal structures of RT complexed with NNRTIs have been obtained. They provide detailed insights in the molecular interaction of the NNRTIs with the amino acids lining the pocket in HIV-1 RT. Due to their unprecedented specificity, the NNRTIs are relatively non-toxic in cell culture, and the most potent compounds reach selectivity indices that exceed 100,000 or more. However, inherent to their high specificity, the NNRTIs easily select for mutant virus strains with several degrees of drug resistance. The first-generation NNRTIs such as nevirapine and delavirdine easily loose their inhibitory potential against mutant virus strains that contain single amino acid mutations in their RT. The second-generation NNRTIs such as efavirenz, capravirine and etravirine [corrected] usually require two or more mutations in the HIV-1 RT before significantly decreasing their antiviral potency. Evidently, it requires a markedly longer time period to obtain significant resistance against second generation NNRTIs. The resistance spectrum of NNRTIs is entirely different from the NRTI resistance spectrum, and, as a rule, NRTI-resistant mutant virus strains keep full sensitivity to the inhibitory effects of NNRTIs, and vice versa NNRTI resistant and mutant virus strains keep full sensitivity to the inhibitory effects of NRTIs. NNRTIs have proven beneficial when included in drug combination (triple or quadruple) therapy, preferably in the presence of protease inhibitors and NRTIs. PMID- 15134550 TI - TSAO compounds: the comprehensive story of a unique family of HIV-1 specific inhibitors of reverse transcriptase. AB - Emergence of drug-resistant viral strains is one of the major milestones and the main cause for the failure of antiretroviral therapy. Combination of different anti-HIV agents has become the standard clinical practice to keep the viral load at low or even undetectable levels and to prevent emergence of virus-drug resistance. Among the human immunodeficiency virus (HIV) reverse transcriptase (RT) inhibitors, the so called nonnucleoside RT inhibitors (NNRTIs) have gained a definitive place in the treatment of HIV infections in combination with nucleoside analogue RT inhibitors (NRTIs) and HIV protease inhibitors (PIs). The virus can be markedly suppressed for a relatively long period of time when exposed to multiple drug combination therapy (highly active antiretroviral therapy, HAART). TSAO derivatives are a peculiar group of highly functionalized nucleosides that belong to the so-called nonnucleoside RT inhibitors (NNRTIs). They exert their unique selectivity for HIV-1 through a specific interaction with the p51 subunit of HIV-1 RT. They are the first small molecules that seem to interfere with the dimerization process of the enzyme. This review covers the work carried out with this unique class of specific inhibitors of HIV-1 reverse transcriptase, including structure activity relationship studies (SAR), its mechanism of action, resistance studies, model of interaction with the enzyme, etc. PMID- 15134551 TI - Structure and function of HIV-1 integrase. AB - HIV-1 integrase is a multidomain enzyme which is required for the integration of viral DNA into the host genome. It is one of three enzymes of HIV, the others being the Reverse Transcriptase and the Protease. It is an attractive target for therapeutic drug design. The enzyme consists of three domains. The N-terminal domain has a His2Cys2 motif which chelates zinc, the core domain has the catalytic DDE motif which is required for its enzymatic activity, and the C terminal domain has an SH3-like fold which binds DNA nonspecifically. We review the structures of various integrase fragments, the core domain with inhibitors bound, and propose a model for DNA binding. PMID- 15134552 TI - HIV-1 integrase: a target for new AIDS chemotherapeutics. AB - Since the beginning of the HIV epidemic almost 70 million people have been infected with HIV. It is estimated that 42 million people are currently living with HIV/AIDS. The spread of HIV continues throughout the world and current estimates indicate that in 2002, 5 million people were newly infected with HIV and 3 million people died. Current treatments employ a combination of therapeutic agents that target the viral reverse transcriptase and protease enzymes and viral entry. However the clinical benefit of these agents is often limited due to issues of regimen compliance, significant side effects, and the emergence of viral strains that are drug resistant. The introduction of novel agents that interfere with alternate stages in the viral life cycle represent potential solutions to these problems. The integration of the HIV genome into the cellular chromosome, a process catalyzed by the viral enzyme integrase, has been shown to be essential for viral replication. Since HIV integrase has no direct cellular counterpart it presents itself as an attractive target for therapeutic intervention. This review summarizes recent and promising developments both in the HIV integrase field and the global quest for therapeutically useful inhibitors of HIV integrase. PMID- 15134553 TI - HIV protease inhibition: limited recent progress and advances in understanding current pitfalls. AB - The identification of HIV-1 protease (HIVp) as a target for therapeutic intervention against AIDS was soon followed by major efforts to understand its substrate specificity, reaction kinetics and three-dimensional structure, both in the free state and in complex with a number of ligands including substrate mimics, products, and inhibitors. On the whole these studies have been extremely successful and have had a major impact on our understanding of ligand-receptor interactions and enzyme inhibition mechanisms. HIVp has also become a paradigm for the development and testing of new drug-design methodologies both in vitro and in silico. Even though thousands of potential HIVp inhibitors exhibiting amazing chemical diversity have been synthesized or identified from natural sources, only a few have turned out to be useful for human therapy. Although the alternative goal of preventing enzyme dimerization has been achieved as a proof of concept, this approach has not yet yielded a clinical candidate. The review covers the general strategies that led to some of the most useful inhibitors, the reasons for our limited success in effectively inhibiting this retroviral target in a clinical setting, current progress with second-generation inhibitors, and new avenues for research. PMID- 15134554 TI - Antiviral profile of HIV inhibitors in macrophages: implications for therapy. AB - Macrophages (M/M) are identified as the second cellular target of HIV and a crucial virus reservoir. M/M are persistently infected cells and not susceptible to the HIV cytophatic effects typical of infected CD4+ T-lymphocytes. HIV replication in M/M is a crucial pathogenetic event during the whole course of the disease. Moreover, the dynamics of HIV-1 replication and cumulative virus production is quite different in M/M and CD4+ T-lymphocytes in the presence or in the absence of antiviral drugs. Thus, for their unique cellular characteristics, the activity of anti-HIV compounds could be different in M/M than in CD4+ T lymphocytes. Indeed, nucleoside analogues inhibitors of HIV-reverse transcriptase (NRTIs) show potent antiviral activity in macrophages, although the limited penetration of these compounds in sequestered body compartments and the scarce phosphorylation ability of macrophages, suggest that a phosphate group linked to NRTIs may confer a greater anti-HIV activity in such cells. The antiviral activity of non-nucleoside reverse transcriptase inhibitors (NNRTIs) in macrophages is similar to that found in CD4-lymphocytes. Interestingly, protease inhibitors (PIs), acting at post-integrational stages of virus replication, are the only drugs able to interfere with virus production and release from macrophages with established and persistent HIV infection. For these reasons, a careful analysis of the distribution of antiviral drugs, and the assessment of their activity in cells of macrophage lineage, represent key factors in the development of therapeutic strategies aimed to the treatment of the HIV-infected patients. PMID- 15134555 TI - The role of the platelet glycoprotein IIb/IIIa in thrombosis and haemostasis. AB - Haemostasis is a finely balanced and complex process ideally initiated only in response to disruption of the vascular endothelium as a means of preventing loss of blood from an injured vessel. Deviations from the ideal can lead to serious disease. Firstly, thrombosis, which arises as a consequence of inappropriate platelet-platelet interactions at a region of vessel damaged by atherosclerosis, can lead to occlusion of the affected vessel as in myocardial infarction or stroke. Secondly, loss of the ability of platelets to form aggregates leads to Glanzmann's thrombasthenia (GT) with a tendency to bleed for prolonged periods following injury. Glycoprotein IIb/IIIa (GPIIb/IIIa) plays a major role in the regulation of platelet adhesion and aggregation during haemostasis. Upon platelet activation by an agonist a signalling process is initiated, termed "inside-out" signalling, which gives rise to conformational changes within GPIIb/IIIa. These conformational changes increase the affinity of the receptor for its primary ligand, fibrinogen. Bound fibrinogen then acts as a bridging molecule facilitating the interaction of adjacent platelets. Upon fibrinogen binding GPIIb/IIIa undergoes further conformational changes and through a process termed "outside-in" signalling the receptor signals in to the platelet ultimately resulting in acceleration of the aggregation process. Qualitative or quantitative abnormalities in GPIIb/IIIa give rise to GT, a recessive bleeding disorder, and analysis of affected individuals has provided invaluable insights into the structure/function relationship of this receptor. Due to its critical role in mediating platelet aggregate formation GPIIb/IIIa has become a primary target for the development of antithrombotic agents. PMID- 15134556 TI - Intravenous glycoprotein IIb/IIIa antagonists: their benefits, problems and future developments. AB - The intravenous Glycoprotein (GP) IIb/IIIa antagonists are potent antiplatelet agents that are particularly effective in patients who undergo percutaneous coronary intervention (PCI). Questions remain about their benefit in the setting of primary PCI, as well as in patients with acute coronary syndromes who do not undergo PCI. The dosing of these drugs is critical to their efficacy and for some agents may not yet be optimized. Differences in the level of platelet inhibition achieved with previous and current dosing strategies of these agents are discussed. In addition, the pharmacology of GPIIb/IIIa antagonists is more complex than initially appreciated. These drugs appear to have partial agonist properties and as a result may be prothrombotic at lower doses. Recent evidence also suggests that at least some of the GPIIb/IIIa antagonists may have anti inflammatory as well as anti-thrombotic activity. Future research should clarify these issues. Because of the observed inter-individual variation in the response to GPIIb/IIIa antagonists, future trials of these agents should also look at individual tailoring of the dose to an optimum level of platelet inhibition. No definite clinical predictors of this inter-individual variation have been identified, but the Pl(A) polymorphism in GPIIIa appears to be associated with an adverse response to treatment with the oral GPIIb/IIIa antagonists in particular. PMID- 15134557 TI - Oral GPIIb/IIIa antagonists: what went wrong? AB - GPIIb/IIIa receptor antagonists block fibrinogen binding to platelets and as a result inhibit platelet aggregation. They are very potent inhibitors due to the critical role fibrinogen binding plays in platelet aggregation. When given intravenously these drugs have been shown to be very effective as adjuvant therapy in percutaneous coronary intervention and in acute coronary syndromes. However, despite being as potent as their intravenous counterparts, all of the oral inhibitors showed no benefit or even increased mortality in clinical trials. There are a number of reasons for their failure. The target was different, chronic treatment to prevent thrombotic events as opposed to short-term treatment to prevent acute events and as a result, different dosing regimens were used. The acute use aims for a high level of inhibition (80-90%) while the chronic use produced lower levels of inhibition. Many of the oral inhibitors had low bioavailability that led to a large peak-trough difference. Most GPIIb/IIIa antagonists have the ability to activate platelets through a GPIIb/IIIa-mediated process. This is known as partial agonism. In the presence of high drug levels, such as during an infusion this is not a problem, however combined with the low trough levels with oral inhibitors this can lead to an increase in platelet aggregation. Other problems include drug-induced conformational changes in GPIIb/IIIa (ligand-regulated binding sites) and possible pharmacogenomics effects in the response to the drugs, in particular the Pl(A) polymorphism in GPIIb/IIIa. By addressing these issues it is possible for a new generation of oral GPIIb/IIIa antagonist to be developed. PMID- 15134558 TI - UR-3216: a new generation oral platelet GPIIb/IIIa antagonist. AB - Various oral platelet GPIIb/IIIa receptor antagonists have undergone clinical investigations, but to date without success. Various factors have been proposed to explain their failure such as low affinity for the receptor, large peak/trough ratio, low bioavailability, partial agonist activity and pro-aggregatory effect. Efforts to discover a truly effective, safe, oral antagonist led to the discovery of UR-3216 (Fig. 1). The active form of UR-3216, UR-2922, possessed a high affinity for the human platelet receptor (K(d) <1 nM) with a slow dissociation rate (k(off)= 90 min) in vitro. UR-2922 induced no ligand-induced binding sites (LIBS) expression or prothrombotic activity in human platelets, distinctly different from orbofiban and other small molecule antagonists. To date, UR-2922 is the only high affinity GPIIb/IIIa antagonist without LIBS expression. In vivo characteristics of UR-3216 showed prolonged duration of efficacy (>24 h) with its favorable pharmacokinetic profile, superior to all the other oral GPIIb/IIIa antagonists. UR-3216 showed high bioavailability, rapid bioconversion to the active form and biliary excretion. UR-3216 is a novel, orally active GPIIb/IIIa antagonist of a new generation, which has substantially improved the crucial compounding factors and will be useful for the treatment of cardiovascular diseases. PMID- 15134559 TI - Novel integrin-specific targets for anti-platelet therapies. AB - Extensive study in integrin research has seen the platelet specific receptor alpha(IIb)beta(3) (Glycoprotein GPIIb/IIIa) under much scrutiny, and provided vast information as to the workings of this integrin within the blood. Glanzmann's thrombasthenia, a rare autosomal recessive bleeding disorder, highlights the vital role played by this receptor in platelet function. Glanzmann's thrombasthenic platelets fail to aggregate due to a lack of surface expression of functional alpha(IIb) or beta(3) on the platelet surface. However, little is known about the precise molecular mechanisms involved in the operation of this receptor on the platelet surface. Clinical trials using intravenous antagonists to this receptor have shown them to be effective anti-thrombotics. However the recent observations that the oral alpha(IIb)beta(3)antagonists have failed to show benefits in the treatment of acute coronary syndromes, and in fact, increase mortality, underscores the necessity for a more complete understanding of alpha(IIb)beta(3) and its functions. A more profound knowledge of the precise nature of the platelet integrin-activation, along with an understanding of its interactions with cellular signaling proteins, will undoubtedly lead to the identification of novel strategies for the effective inhibition of platelet integrin function. PMID- 15134560 TI - Oxygen, reactive oxygen species and tissue damage. AB - The diatomic molecule of oxygen contains two uncoupled electrons and can therefore undergo reduction, yielding several different oxygen metabolites, which are collectively called Reactive Oxygen Species or ROS. They are invariably produced in aerobic environments through a variety of mechanisms, which include electron "leakage" during biologic oxidations, action of flavin dehydrogenases and specific membrane associated secretion, as well as by physical activation of oxygen by irradiation, e.g. UV sun-light. Organisms have developed efficient protective mechanisms against excessive accumulation of ROS, which include superoxide anion, hydrogen peroxide and hydroxyl radical, since all these metabolites are highly reactive and affect almost every kind of organism, either directly or through conversion into other derivatives, notably NO-derived radicals or RNS. Depending on their tissue concentration they can either exert beneficial physiologic effects (control of gene expression and mitogenesis) or damage cell structures, including lipids and membranes, proteins and nucleic acids, leading to cell death. In this brief overview we summarize the present state-of-the-art, restricting the discussion to the role of ROS in physiology and pathology, not taking into account RNS. Discussion will focus on basic chemical and biochemical features of ROS, underlining how ROS can promote severe diseases, including neoplastic, cardiovascular and neurodegenerative diseases. This brief discussion should clarify the present huge interest in ROS, in the perspective to develop new and specific therapeutic approaches. PMID- 15134561 TI - Biochemical aspects of nitric oxide. AB - Nitric oxide (NO), a free radical molecule, produced by NO synthase (NOS) in the body exerts a number of pathophysiological actions due to its chemical reactivity. Low amounts of NO (nM) normally produced by constitutive NOS play a critical role in different physiological events such as vasodilation and neurotransmission. Higher amounts of NO ( micro M) locally and spatially produced by inducible NOS during inflammation act as double-edged sword exerting either beneficial or detrimental effects. Recently, new vision on the biological role of NO has been proposed based on the possible cross-talk between constitutive and inducible NOS. Accordingly, normally produced low amounts of NO may be involved in the regulation of NF-kappaB activation and successively the expression of inducible NOS. Under normal conditions NF-kappaB activation is suppressed by low amounts of NO. Under conditions in which massive amounts of NO produced by inducible NOS act detrimentally, NO-elicited down-regulation of NF-kappaB activation is compromised due to the drop in NO at the early phase of inflammation caused by inactivation of constitutive NOS. Any treatment which counterparts the drop in NO, therefore, may present a new approach either in preventing or in treating inflammatory diseases. PMID- 15134562 TI - New pathways for reactive oxygen species generation in inflammation and potential novel pharmacological targets. AB - Oxidative agents are generated in large amounts during inflammation. These highly reactive intermediates interact with several extracellular and intracellular molecules and with each other, thus generating a complex network of responses culminating in an outcome that may be detrimental or beneficial for the host. Alongside with the well known systems involved in production of reactive oxygen species or reactive nitrogen species, such as the NADPH oxidase or the nitric oxide synthase, novel enzymatic pathways have been discovered. This has unveiled new targets and functions for oxidant species, and has prompted the development of innovative anti-inflammatory drugs. In the new integrate scenario stemming from these studies oxidant species are increasingly recognized as true messengers, and even their toxic effects are viewed as the result of the perversion of an otherwise physiological extra/intra cellular signaling. PMID- 15134563 TI - Linking inflammation to cell cycle progression. AB - Risk of gastrointestinal cancers is closely related to increased levels of oxidants in the balance between oxidant and anti-oxidant agents. A possible explanation of this epidemiological observation is the local loss of the epithelial barrier function with a focal inflammatory response. Accordingly, chronic inflammatory diseases represent well-known risk factors for cancer and, on the other hand, it is known that anti-inflammatory agents, demulcents and antioxidants markedly inhibit the development of colon cancer in animal models as well in humans. At molecular level a key role in the process that link inflammation to cellular transformation seems to be played by activation of Cyclooxygenase-2 (COX-2) together with production of Reactive Oxygen Intermediate (ROI). Both these events have been strictly linked with cell proliferation and transformation, although the intracellular pathways involved in these processes are still not completely understood. The uncontrolled proliferation, which is a landmark of cellular transformation, is accompanied by the deregulation of proteins involved in the control of cell cycle checkpoints. Altered expression and function of cyclooxygenase and nitric oxide synthase seem to influence, among others, the expression of proteins involved in the regulation of cell cycle progression. Similarly, anti-inflammatory and antioxidant agents may also act on the expression and function of several cell cycle regulating proteins. Understanding the mechanisms by which chronic inflammation contributes to genetic and epigenetic changes involved in the regulation of critical cell cycle checkpoints may help to develop more and more specific treatment strategies for reducing malignant transformation of these inflammatory diseases. PMID- 15134564 TI - Pharmacology of nitric oxide: molecular mechanisms and therapeutic strategies. AB - NO is considered to be an ubiquitous endogenous system which takes part in body's homeostatic regulations and in pathological events. NO derives from a) the actions of enzymes, the NO Synthases (NOS), which are constitutives (endothelial NOS (eNOS) and nervous NOS (nNOS)) and generate small amounts of NO and have homeostatic functions: and b) from the actions of inducible NOS (iNOS), which generate large amounts of NO and exert protective actions against noxious agents but also toxic effects (e.g. inhibition of enzymes) through the production of peroxynitrite (ONOO-). Modulation of the L-Arg/NO system may be used to obtain favourable therapeutic results, either by promoting (e.g. with NO donors) or by reducing (e.g. with NOS inhibitors) the production of NO. The present chapter will consider two approaches and four groups of potential therapeutic agents: 1) The stimulation of NO production with; a) agents which improve the efficiency of the Kallikrein-Kinin System; b) NO donors. 2) The reduction of excessive NO production with: a) inhibitors of NO Synthases; b) agents that reduce the formation of reactive nitrogen/ oxygen species (RNS / ROS). PMID- 15134565 TI - The antioxidants and pro-antioxidants network: an overview. AB - Living beings have evolved over the past two billion years through adaptation, to an increasing atmospheric oxygen concentration, by both taking advantage of oxygen activating function and developing a complex control network. In these regards, potentially damaging species (reactive oxygen, nitrogen and chlorine species) arise as by-products of metabolism and also work as physiological mediators and signalling molecules. Oxidative stress may be an important factor in numerous pathological conditions, i.e. infection if micronutrients are deficient. Levels of these species are controlled by the antioxidant defence system, which is composed by antioxidants and pro-antioxidants. Several components of this system are micronutrients (e.g. vitamins C and E), are dependent upon dietary micronutrients (e.g. CuZn and Mn superoxide dismutase) or are produced by specific endogenous pathways. The antioxidant defences act, to control levels of these species, as a coordinated system where deficiencies in one component may affect the efficiency of the others. In this network some of the components act as direct antioxidants whereas others act indirectly (pro antioxidants) either by modulation of direct agents or by regulation of the biosynthesis of antioxidant proteins. Thus, entities usually not considered as antioxidants, also act efficiently counteracting damaging effects of oxidative species. In this contest, the design of new molecules that take into account synergistic interactions among different antioxidants, could be useful both to address mechanistic studies and to develop possible therapeutic agents. In this review the principal categories of antioxidants and pro-antioxidants that goes from vitamins through phyto-derivatives to minerals, are critically reviewed, with particular emphasis on structure-function considerations, together with the perspective opened, in the design of possible therapeutic agents, by the antioxidants interplay. PMID- 15134566 TI - Oxidative stress in essential hypertension. AB - A major cause for endothelial dysfunction in essential hypertension is decreased availability of nitric oxide (NO). Impairment in NO bioavailability is likely to be the consequence of multiple mechanisms affecting NO synthesis as well as NO breakdown. An alteration in the redox balance in endothelial cells leads to increased superoxide anion production and oxidative stress. This in turn not only exerts negative effects on vascular tone, but is also able to activate important mechanisms (such as platelet activity, leukocyte adhesion, vascular smooth muscle cell proliferation and expression of adhesion molecules) with an established central role in the pathogenesis of hypertensive target organ damage. As a consequence, a drug therapy able to restore NO availability in essential hypertensive patients would probably exert additional benefits, as compared to blood pressure lowering per se, in terms of prevention of target organ damage and improved prognosis of these patients. Unfortunately, as of today only the antagonists of the renin-angiotensin system and the calcium-channel blockers have shown some ability in this respect, whereas no longitudinal intervention study has been undertaken, so far, to prove that the restoration of NO bioavailability through an antihypertensive treatment may confer additional prognostic advantage to essential hypertensive patients. PMID- 15134567 TI - Oxidative stress during myocardial ischaemia and heart failure. AB - Oxidative stress is a condition in which oxidant metabolites exert their toxic effect because of an increased production or an altered cellular mechanism of protection. The heart needs oxygen avidly and, although it has powerful defence mechanisms, it is susceptible to oxidative stress, which occurs, for instance, during post-ischaemic reperfusion. Ischaemia causes alterations in the defence mechanisms against oxygen free radicals, mainly a reduction in the activity of mitochondrial superoxide dismutase and a depauperation of tissue content of reduced glutathione. At the same time, production of oxygen free radicals increases in the mitochondria and leukocytes and toxic oxygen metabolite production is exacerbated by re-admission of oxygen during reperfusion. Oxidative stress, in turn, causes oxidation of thiol groups and lipid peroxidation leading first to reversible damage, and eventually to necrosis. In man, there is evidence of oxidative stress (determined by release of oxidised glutathione in the coronary sinus) during surgical reperfusion of the whole heart, or after thrombolysis, and it is related to transient left ventricular dysfunction or stunning. Data on oxidative stress in the failing heart are scant. It is not clear whether the defence mechanisms of the myocyte are altered or whether the production of oxygen free radicals is increased, or both. Recent data have shown a close link between oxidative stress and apoptosis. Relevant to heart failure is the finding that tumour necrosis factor, which is found increased in failing patients, induces a rapid rise in intracellular reactive oxygen intermediates and apoptosis. This series of events is not confined to the myocytes, but occurs also at the level of endothelium, where tumour necrosis factor causes expression of inducible nitric oxide synthase, production of the reactive radical nitric oxide, oxidative stress and apoptosis. It is therefore, possible that the immunological response to heart failure results in endothelial and myocyte dysfunction through oxidative stress mediated apoptosis. Clarification of these mechanisms may lead to novel therapeutic strategies. PMID- 15134568 TI - Radiolabeled tracers for imaging of tumor angiogenesis and evaluation of anti angiogenic therapies. AB - A variety of therapeutic strategies in oncology are focused on the inhibition of tumor-induced angiogenesis. Thus, there is a keen interest in methods which allow non-invasive monitoring of molecular targets involved in angiogenesis which would support information for planning and controlling corresponding therapies. Moreover, such techniques would provide an insight into the formation of new sprouting blood vessels, the involved processes and regulatory mechanisms in patients. At the moment, development of radiotracer based techniques is mainly concentrated on three different targets which include peptidic and non-peptidic alpha v beta 3-integrin binding antagonists, matrix metalloproteinase inhibitors and single chain anti-fibronectin antibody fragments. Development of radiolabeled MMP inhibitors is based on either the decapeptide Cys-Thr-Thr-His-Trp-Gly-Phe-Thr Leu-Cys resulting from a phage display library or small molecular weight compounds. The in vitro data for these tracers are very promising. However, more detailed in vivo data are necessary to evaluate the potency of MMP-inhibitors for in-vivo imaging. The radiolabelled anti-ED-B single chain antibody fragment scFv L-19 shows selective accumulation in the tumor vasculature in a murine tumour model. In a first patient study a selective localisation of the (123)I-labeled tracer in lesions of different tumours was found. On the basis of the lead structure cyclo(-Arg-Gly-Asp-dPhe-Val) a variety of different radiolabeled RGD peptides has been developed for the non-invasive determination of the alpha v beta 3 expression. These developments include peptides labeled with minimum structural alteration, peptide carbohydrate conjugates, peptidomimetics based on the RGD-structure as well as heterodimeric, homodimeric and homotetrameric ligand systems. Many of the tracers show high alpha v beta 3-affinity and selectivity in vitro and receptor selective tumour accumulation with high image contrast in different murine tumour models. Further studies have to demonstrate that this approach can be translated to clinical settings allowing visualisation of alpha v beta 3-positive tumours and alpha v beta 3 expression during tumour-induced angiogenesis in patients. PMID- 15134569 TI - Recent advances in the imaging of programmed cell death. AB - A deficiency or an excess of programmed cell death (apoptosis) is an integral component of autoimmune disorders, organ and bone marrow transplant rejection, and cancer. A technique to image programmed cell death would be useful in the development of drugs to treat these and others diseases, and to monitor the effectiveness of therapy. The most widely studied agent for the in vivo study of apoptosis is radiolabeled annexin V, an endogenous protein labeled with technectium-99m, now undergoing clinical trials in both Europe and the United States. While annexin V has been studied extensively in humans the precise mechanism(s) of uptake of this agent in vivo is unclear and needs further study. Other agents are also underdevelopment including radiolabeled forms of Z-VAD.fmk, a potent inhibitor of the enzymatic cascade intimately associated with apoptosis. MR imaging techniques and tracers also hold promise as methods to monitor apoptotic cell death. In this article we will review these and other imaging technologies for the non-invasive imaging of cell death. The mechanism(s) and latest data on the conditions in which cellular stress and early apoptosis occur will also be discussed in detail including potential new strategies for the targeting and novel therapeutic interventions of tissues and organs undergoing stress or apoptosis when cell salvage is still possible. PMID- 15134570 TI - Imaging beta-amyloid plaques and neurofibrillary tangles in the aging human brain. AB - The development of radioligands to image beta-amyloid (A beta) plaques and neurofibrillary tangles (NFTs) in vivo in the aging human brain is an important and active area of radiopharmaceutical design. When used in combination with positron emission tomography (Pet) or single photon emission computed tomography (spect), amyloid-imaging tracers could facilitate the evaluation of the efficacy of anti-amyloid therapies currently under intense development by many major pharmaceutical companies throughout the world. Amyloid-imaging agents could also serve as surrogate markers in early diagnosis and neuropathogenesis studies of Alzheimer's disease and other aging-related neurodegenerative disorders. In this review article, the design and biological evaluation of amyloid-imaging agents are discussed. The structures of these agents vary from large proteins and peptides such as radiolabeled A beta peptides and monoclonal antibodies to small molecules derived from Congo red, Chrysamine-G, thioflavin-T, and Acridine Orange. In vitro studies indicate that amyloid plaques contain multiple binding sites that can accommodate structurally diverse compounds, providing flexibility for radiopharmaceutical design of amyloid imaging agents. Compared to large biomolecules, small molecule radiotracers are often readily accessible through chemical synthesis and can display superior brain permeability. Several small molecule amyloid-imaging radioligands display high binding affinities to A beta and sufficient brain penetration for imaging studies. Recent studies demonstrate the feasibility of imaging amyloid plaques in vivo in human subjects with PET. Imaging NFTs, separately or in concert with A beta plaques, is not as far advanced as imaging A beta plaques and remains to be fully characterized and demonstrated. PMID- 15134571 TI - PET Studies on P-glycoprotein function in the blood-brain barrier: how it affects uptake and binding of drugs within the CNS. AB - Permeability of the blood-brain barrier (BBB) is one of the factors determining the bioavailability of therapeutic drugs. The BBB only allows entry of lipophilic compounds with low molecular weights by passive diffusion. However, many lipophilic drugs show negligible brain uptake. They are substrates for transporters such as P-glycoprotein (P-gp), multidrug-resistance associated protein (MRP) and organic anion transporting polypeptides (OATPs). The action of these carrier systems results in rapid efflux of xenobiotics from the central nervous system (CNS). Classification of candidate drugs as substrates or inhibitors of such carrier proteins is of crucial importance in drug development. Positron emission tomography (PET) can play an important role in the screening process by providing in vivo information, after the putative drug has passed in vitro tests. Although radiolabeled probes for MRP and OATP function are not yet available, many radiotracers have been prepared to study P-glycoprotein function in vivo with PET. These include alkaloids ((11)C-colchicine), antineoplastic agents ((11)C-daunorubicin, (18)F-paclitaxel), modulators of L-type calcium channels ((11)C-(+/-)verapamil, (11)C-R(+)-verapamil), beta-adrenoceptor antagonists ((11)C-(S)-carazolol, (18)F-(S)-1'-fluorocarazolol, (11)C carvedilol), serotonin 5-HT(1A) receptor antagonists ((18)F-MPPF), opioid receptor antagonists ((11)C-loperamide, (11)C-carfentanyl), and various (64)Cu labeled copper complexes. Studies in experimental animals have indicated that it is possible to assess P-glycoprotein function in the BBB and its effect on the uptake and binding of drugs within the intact CNS, using suitable P-gp modulators labeled with positron emitters. Provided that radiopharmaceuticals (and P-gp modulators) can be developed for human use, several exciting fields of study may be explored, viz. (i) direct evaluation of the effect of modulators on the cerebral uptake of therapeutic drugs; (ii) assessment of mechanisms underlying drug resistance in epilepsy; (iii) examination of the role of the BBB in the pathophysiology of neurodegenerative and affective disorders; and (iv) exploration of the relationship between polymorphisms of transporter genes and the pharmacokinetics of test compounds within the CNS. PMID- 15134572 TI - Acetylcholinesterase imaging: its use in therapy evaluation and drug design. AB - Several cholinesterase (ChE) inhibitors have been labeled with carbon-11 for visualizing binding sites on acetylcholinesterase (AChE) by positron emission tomography (PET). Following intravenous injection of 1,2,3,4-tetrahydro-9 [(11)C]methylaminoacridine or [(11)C]donepezil, however, the radioactivity distribution does not reflect the regional distribution of AChE in the brain of animals, probably because these compounds have high non-specific binding and/or other specific binding sites in vivo in the brain. PET studies with [(11)C]physostigmine and [(11)C]CP-126,998 in the brain of healthy subjects have shown a radioactivity distribution corresponding to the regional distribution of AChE activity measured in postmortem human brains. These radiotracers may be useful for measuring the occupancy of binding sites on AChE by AChE inhibitors, and for investigating the cerebral pharmacokinetics of such therapeutic drugs. An alternative approach to map AChE is the use of acetylcholine analogue substrates. We have developed N-methylpiperidinyl esters labeled with carbon-11 for quantitative measurement of AChE activity. Currently, two N [(11)C]methylpiperidine esters, N-[(11)C]methylipiperidin-4-ylacetate (MP4A) and N-[(11)C]methylpiperidin-4-yl propionate (MP4P or PMP), have been used for clinical studies of Alzheimer's disease and other neurodegenerative diseases. Both [(11)C]MP4A- and [(11)C]MP4P-PET have demonstrated not only the reduction of AChE activity in the cerebral cortex of patients with Alzheimer's disease (AD) but also the inhibitory effects of donepezil and rivastigmine on AChE activity in the brain of AD patients. AChE imaging should prove useful for therapeutic monitoring of the effects of ChE inhibitors, including determination of the appropriate clinical doses of newly developed compounds, and can thus prompt the development of novel drugs targeting AChE. PMID- 15134573 TI - PET imaging of beta-adrenoceptors in human brain: a realistic goal or a mirage? AB - Beta-adrenoceptors are predominantly located in the cerebral cortex, nucleus accumbens and striatum. At lower densities, they are also present in amygdala, hippocampus and cerebellum. Beta-2 sites regulate glial proliferation during ontogenic development, after trauma and in neurodegenerative diseases. The densities of beta-1 adrenoceptors are changed by stress, in several mood disorders (depression, excessive hostility, schizophrenia) and during treatment of patients with antidepressants. A technique for beta-adrenoceptor imaging in the human brain is not yet available. Although 24 (ant)agonists have been labeled with either (11)C or (18)F and some of these are successful myocardial imaging agents, only two (S-1'-(18)F-fluorocarazolol and S-1'-(18)F-fluoroethylcarazolol) could actually visualize beta-adrenoceptors within the central nervous system. Unfortunately, these radiopharmaceuticals showed a positive Ames test. They may be mutagenic and cannot be employed for human studies. Screening of more than 150 beta-blockers described in the literature yields only two compounds (exaprolol and L643,717) which can still be radiolabeled and evaluated for beta-adenoceptor imaging. However, other imaging techniques could be examined. Cerebral beta adrenoceptors might be labeled after temporary opening of the blood-brain barrier (BBB) and simultaneous administration of a hydrophilic ligand such as S-(11)C CGP12388. Another approach to target beta-adrenoceptor ligands to the CNS is esterification of a myocardial imaging agent (such as (11)C-CGP12177), resulting in a lipophilic prodrug which can cross the BBB and is split by tissue esterases. BBB opening is not feasible in healthy subjects, but the prodrug approach may be successful and deserves to be explored. PMID- 15134574 TI - Diagnostic and therapeutic applications of recombinant antibodies: targeting the extra-domain B of fibronectin, a marker of tumor angiogenesis. AB - Angiogenesis, the sprouting of new blood vessels from preexisting ones, is a phenomenon associated to several human pathologies, including different potentially blinding ocular disorders, rheumatoid arthritis, and cancer. Indeed, ongoing angiogenesis is a characteristic feature of the majority of aggressive solid tumors, and is also a pre-requisite for the progression towards the metastatic phenotype. One established marker of angiogenesis is represented by an isoform of the oncofoetal fibronectin (FN), containing an additional domain inserted by alternative splicing of the FN pre-mRNA and called extra-domain B (ED B). This isoform has been found to be present almost exclusively in the modified extra-cellular matrix surrounding newly-formed blood vessels in tumors (and other animal models of ocular pathologies), being completely absent from the normal vasculature in adult organs. This article reviews the recombinant antibodies raised against ED-B and the different methodologies used for the generation of these antibodies. Moreover, new diagnostic and therapeutic applications based on the delivery of bioactive molecules to tumor blood vessels by means of ED-B targeting will be discussed. PMID- 15134575 TI - Coupling factors in macromolecular type-IV secretion machineries. AB - Type IV secretion systems (T4SSs) are bacterial multiprotein organelles specialised in the transfer of (nucleo)protein complexes across cell membranes. They are essential for conjugation, bacterial-induced tumour formation in plant cells, as observed in Agrobacterium, toxin secretion, like in Bordetella and Helicobacter, cell-to-cell translocation of virulence factors, and intracellular activity of mammalian pathogens like Legionella. By enabling conjugative DNA delivery, these systems contribute to the spread of antibiotic resistance genes among bacteria. These translocons are made up by 10-15 proteins that are analogous to Vir proteins of Agrobacterium and traverse both membranes and the periplasmic space in between in Gram-negative bacteria. Their secretion substrates range from single-stranded DNA/protein complexes to multicomponent toxins and they are assisted by integral inner-membrane coupling factors, the multimeric type-IV coupling proteins (T4CPs), to connect the macromolecular complexes to be transferred with the secretory conduit. To do so, these T4CPs may be required to localise close to the secretion machinery within the donor cell. The T4CP structural prototype is the hexameric protein TrwB of Escherichia coli conjugative plasmid R388, closely related to Agrobacterium VirD4 protein. It is responsible for coupling the relaxosome with the DNA transport apparatus during cell mating. T4CP family members are related to SpoIIIE/FtsK proteins, essential for DNA pumping during sporulation and cell division. These features suggest possible mechanisms for conjugal T4CP function: as a simple coupler between two molecular machines, as a rotating device to pump DNA through the type-IV transport pore, or as a DNA injector, whereby its central channel would function as part of the transport pore. PMID- 15134576 TI - Elicitor effects on Penicillium chrysogenum morphology in submerged cultures. AB - Changes in morphology and sporulation were investigated in liquid cultures of Penicillium chrysogenum P2 supplemented with carbohydrate oligosaccharides. Sodium alginate and locust-bean (Ceratonia siliqua) gum-derived oligosaccharides were used as elicitors. Spore germination was inhibited by the addition of OG (oligoguluronate) elicitor (30% inhibition when compared with control). Addition of any of the elicitors to stirred-tank cultures increased hyphal-tip numbers, clump area and spore counts. MO (mannan oligosaccharide) had the greatest effect on the parameters studied, followed by OM (oligomannuronate) and OG. Hyphal-tip numbers increased by 19, 29 and 47% with OG, OM and MO respectively. Average clump area in the presence of OG, OM and MO increased by 23, 32 and 59% respectively. A notable increase in spore numbers was observed in all the supplemented stirred-tank reactor cultures, with MO showing the highest enhancement. PMID- 15134577 TI - Altered calcium signaling in platelets from nitric oxide-deficient hypertensive rats. AB - BACKGROUND: In the present study we have analyzed the mechanisms of calcium entry and mobilization in platelets obtained from rats chronically treated with the nitric oxide synthesis inhibitor, N-nitro L-arginine methyl ester [L-NAME, 40 mg/kg/day, 5 days). The platelets were obtained the day of the experiment, washed and loaded with fura-2. The intracellular calcium levels were determined in suspension of cells by means of fluorescence spectroscopy. RESULTS: Basal calcium levels were always elevated in the platelets of the L-NAME-treated rats, both in the presence and in the absence of extracellular calcium. The administration of thrombin in the absence and in the presence of extracellular calcium induced important elevations in calcium levels that were always of greater magnitude in the platelets of the L-NAME-treated rats than in those of the controls. The addition of calcium to thapsigargin-treated platelets produced a massive elevation in calcium levels in both groups that was significantly greater in the platelets obtained from the hypertensive rats than in those of the controls. CONCLUSIONS: It is concluded that the arterial hypertension induced by the reduction of nitric oxide alters the regulation of platelet calcium levels so that elevated baseline levels and calcium entry and mobilization are enhanced. This could be the result of direct or indirect effects of the lack of nitric oxide synthesis in platelets or in other tissues. PMID- 15134578 TI - Human apoB contributes to increased serum total apo(a) level in LPA transgenic mice. AB - BACKGROUND: The Lp(a) lipoprotein (Lp(a)) consists of the polymorphic glycoprotein apolipoprotein(a) (apo(a)), which is attached by a disulfide bond to apolipoprotein B (apoB). Apo(a), which has high homology with plasminogen, is present only in primates and hedgehogs. However, transgenic mice and rabbits with high serum apo(a) levels exist. Liver is the main site for apo(a) synthesis, but the site of removal is uncertain. To examine differences between transgenic mice expressing the LPA gene and mice capable of forming Lp(a) particles, LPA-YAC transgenic mice and hAPOB transgenic mice were crossed and their offspring examined. RESULTS: Comparison of LPA-YAC with LPA-YAC/hAPOB transgenic mice showed that LPA-YAC/hAPOB transgenic mice have higher serum total apo(a) and total cholesterol level than mice lacking the hAPOB gene. However, hepatic apo(a) mRNA level was higher in LPA-YAC transgenic mice than in LPA-YAC/hAPOB transgenic mice. Feeding of a high-cholesterol/high-fat diet to male LPA-YAC transgenic mice with or without the hAPOB gene resulted in reduced serum total apo(a) and hepatic apo(a) mRNA level. CONCLUSION: In conclusion, the higher serum total apo(a) level in LPA-YAC/hAPOB transgenic mice than in LPA-YAC transgenic mice is not caused by increased apo(a) synthesis. Lower hepatic apo(a) mRNA level in LPA-YAC/hAPOB than in LPA-YAC transgenic mice may suggest that the increase in total apo(a) level is a result of apo(a) accumulation in serum. Furthermore, observed higher serum total cholesterol level in LPA-YAC/hAPOB transgenic mice than either in wild type or LPA-YAC transgenic mice may further suggest that human APOB transgenicity is a factor that contributes to increased serum total apo(a) and cholesterol levels. Our results on reduced serum total apo(a) and hepatic apo(a) mRNA levels in HCHF fed male LPA-YAC transgenic mice confirm earlier findings in females, and show that there are no sex difference in mechanisms for lowering apo(a) level in response to HCHF feeding. PMID- 15134579 TI - Positive pleiotropic effects of HMG-CoA reductase inhibitor on vitiligo. AB - BACKGROUND: HMG-CoA reductase inhibitors (statins) are commonly used in medicine to control blood lipid disorder. Large clinical trials have demonstrated that statins greatly reduces cardiovascular-related morbidity and mortality in patients with and without coronary artery disease. Also, the use of HMG-CoA reductase inhibitors has been reported to have immunosuppressive effects. CASE PRESENTATION: We describe an unusual case of regression of vitiligo in a patient treated with high dose simvastatin. The relation between simvastatin and regression of vitiligo in this case report may be related to the autoimmune pathophysiology of the disease. CONCLUSION: This unexpected beneficial impact provides another scientific credence to the hypothesis that immune mechanisms play a role in the development of vitiligo and that the use of statins as immuno modulator could be of use not only for treatment relative to organ transplant but in other pathologies such as vitiligo. PMID- 15134580 TI - Is NHS dentistry in crisis? 'Traffic light' maps of dentists distribution in England and Wales. AB - BACKGROUND: 'Traffic light' (red-yellow-green) maps are potentially powerful tools for 'at a glance' problem detection, for optimising resource allocation/reallocation, setting priorities, and targeting interventions to areas most in need. The maps can be also used for administrative area comparisons and performance monitoring over time. Interactive Web versions of the maps can be generated with many handy features to further empower organisations and decision makers. Methodological issues to consider when creating 'traffic light' maps include hue thresholding, data timeliness and stability of administrative boundaries. RESULTS: We used 'traffic light' maps to study the distribution of dentists per 1,000 population in all 304 English Primary Care Trusts (PCTs) and 22 Welsh Local Health Boards (LHBs) using datasets of dentist numbers per PCT (as at 31 December 2002) and LHB (as at 26 February 2004) from the Dental Practice Board, and 2001 Census population figures for PCTs and LHBs from the Office for National Statistics. The overall NHS dentists per 1,000 population figures for England (0.374) and Wales (0.359) are low compared to many other countries, with less than 0.3 dentist per 1,000 people available to 24.1% of the total population of England (81 PCTs or 26.6% of all PCTs) and 26.1% of the total population of Wales (6 LHBs or 27.3% of all LHBs). A general shortage of NHS dentists can be observed at a glance across England and Wales on all the 'traffic light' maps in our study, even on those using a more "tolerant" classification and an additional orange-yellow class. The distribution of NHS dentists in England and Wales was also found to be not uniform, with some PCTs/LHBs, especially those located in some of the deprived or less populated urban and rural communities, suffering significantly more shortage of dentists than others (see http://healthcybermap.org/PCT/dentists/). These results confirm recent media reports of a shortage of NHS dentists in various parts of England and Wales. CONCLUSION: Suitable programmes are urgently needed to increase the numbers of NHS dentists across England and Wales. We have included a set of recommendations to dental health policymakers and planners, in addition to ideas for further work. PMID- 15134581 TI - Prevalence and antimicrobial susceptibilities of bacteria isolated from blood cultures of hospitalized patients in the United States in 2002. AB - BACKGROUND: Bloodstream infections are associated with significant patient morbidity and mortality. Antimicrobial susceptibility patterns should guide the choice of empiric antimicrobial regimens for patients with bacteremia. METHODS: From January to December of 2002, 82,569 bacterial blood culture isolates were reported to The Surveillance Network (TSN) Database-USA by 268 laboratories. Susceptibility to relevant antibiotic compounds was analyzed using National Committee for Clinical Laboratory Standards guidelines. RESULTS: Coagulase negative staphylococci (42.0%), Staphylococcus aureus (16.5%), Enterococcus faecalis (8.3%), Escherichia coli (7.2%), Klebsiella pneumoniae (3.6%), and Enterococcus faecium (3.5%) were the most frequently isolated bacteria from blood cultures, collectively accounting for >80% of isolates. In vitro susceptibility to expanded-spectrum beta-lactams such as ceftriaxone were high for oxacillin susceptible coagulase-negative staphylococci (98.7%), oxacillin-susceptible S. aureus (99.8%), E. coli (97.3%), K. pneumoniae (93.3%), and Streptococcus pneumoniae (97.2%). Susceptibilities to fluoroquinolones were variable for K. pneumoniae (90.3-91.4%), E. coli (86.0-86.7%), oxacillin-susceptible S. aureus (84.0-89.4%), oxacillin-susceptible coagulase-negative staphylococci (72.7 82.7%), E. faecalis (52.1%), and E. faecium (11.3%). Combinations of antimicrobials are often prescribed as empiric therapy for bacteremia. Susceptibilities of all blood culture isolates to one or both agents in combinations of ceftriaxone, ceftazdime, cefepime, piperacillin-tazobactam or ciprofloxacin plus gentamicin were consistent (range, 74.8-76.3%) but lower than similar beta-lactam or ciprofloxacin combinations with vancomycin (range, 93.5 96.6%). CONCLUSION: Ongoing surveillance for antimicrobial susceptibility remains essential, and will enhance efforts to identify resistance and attempt to limit its spread. PMID- 15134583 TI - Efficacy of pyrimethamine-sulfadoxine in young children with uncomplicated falciparum malaria in rural Burkina Faso. AB - The efficacy of pyrimethamine-sulfadoxine in the treatment of uncomplicated falciparum malaria in young children of a malaria holoendemic area in rural Burkina Faso is reported. Of 28 children treated with a standard single dose of pyrimethamine-sulfadoxine and followed-up over 14 days, only one Late Treatment Failure and four Late Parasitological Failures were observed, all with low-grade parasitaemia. In this area of very restricted use of pyrimethamine-sulfadoxine, the drug appears to be still sufficiently effective in the treatment of malaria. These findings provide further evidence for the justification of continued use of pyrimethamine-sulfadoxine as a second-line treatment for malaria in Burkina Faso. PMID- 15134582 TI - Homocysteine and reactive oxygen species in metabolic syndrome, type 2 diabetes mellitus, and atheroscleropathy: the pleiotropic effects of folate supplementation. AB - Homocysteine has emerged as a novel independent marker of risk for the development of cardiovascular disease over the past three decades. Additionally, there is a graded mortality risk associated with an elevated fasting plasma total homocysteine (tHcy). Metabolic syndrome (MS) and type 2 diabetes mellitus (T2DM) are now considered to be a strong coronary heart disease (CHD) risk enhancer and a CHD risk equivalent respectively. Hyperhomocysteinemia (HHcy) in patients with MS and T2DM would be expected to share a similar prevalence to the general population of five to seven percent and of even greater importance is: Declining glomerular filtration and overt diabetic nephropathy is a major determinant of tHcy elevation in MS and T2DM. There are multiple metabolic toxicities resulting in an excess of reactive oxygen species associated with MS, T2DM, and the accelerated atherosclerosis (atheroscleropathy). HHcy is associated with an increased risk of cardiovascular disease, and its individual role and how it interacts with the other multiple toxicities are presented.The water-soluble B vitamins (especially folate and cobalamin-vitamin B12) have been shown to lower HHcy. The absence of the cystathionine beta synthase enzyme in human vascular cells contributes to the importance of a dual role of folic acid in lowering tHcy through remethylation, as well as, its action of being an electron and hydrogen donor to the essential cofactor tetrahydrobiopterin. This folate shuttle facilitates the important recoupling of the uncoupled endothelial nitric oxide synthase enzyme reaction and may restore the synthesis of the omnipotent endothelial nitric oxide to the vasculature. PMID- 15134584 TI - Thermus thermophilus genome analysis: benefits and implications. AB - The genome sequence analysis of Thermus thermophilus HB27, a microorganism with high biotechnological potential, has recently been published. In that report, the chromosomal and the megaplasmid sequence were compared to those of other organisms and discussed on the basis of their physiological and metabolic features. Out of the 2,218 putative genes identified through the large genome sequencing project, a significant number has potential interest for biotechnology. The present communication will discuss the accumulating information on molecules participating in fundamental biological processes or having potential biotechnological importance. PMID- 15134585 TI - Deprived children or deprived neighbourhoods? A public health approach to the investigation of links between deprivation and injury risk with specific reference to child road safety in Devon County, UK. AB - BACKGROUND: Worldwide, injuries from road traffic collisions are a rapidly growing problem in terms of morbidity and mortality. The UK has amongst the worst records in Europe with regard to child pedestrian safety. A traditional view holds that resources should be directed towards training child pedestrians. In order to reduce socio-economic differentials in child pedestrian casualty rates it is suggested that these should be directed at deprived children. This paper seeks to question whether analysis of extant routinely collected data supports this view. METHODS: Routine administrative data on road collisions has been used. A deprivation measure has been assigned to the location where a collision was reported, and the home postcode of the casualty. Aggregate data was analysed using a number of epidemiological models, concentrating on the Generalised Linear Mixed Model. RESULTS: This study confirms evidence suggesting a link between increasing deprivation and increasing casualty involvement of child pedestrians. However, suggestions are made that it may be necessary to control for the urban nature of an area where collisions occur. More importantly, the question is raised as to whether the casualty rate is more closely associated with deprivation measures of the ward in which the collision occurred than with the deprivation measures of the home address of the child. CONCLUSION: Conclusions have to be drawn with great caution. Limitations in the utility of the officially collected data are apparent, but the implication is that the deprivation measures of the area around the collision is a more important determinant of socio economic differentials in casualty rates than the deprivation measures of the casualties' home location. Whilst this result must be treated with caution, if confirmed by individual level case-controlled studies this would have a strong implication for the most appropriate interventions. PMID- 15134586 TI - Signaling of angiotensin II-induced vascular protein synthesis in conduit and resistance arteries in vivo. AB - BACKGROUND: From in vitro studies, it has become clear that several signaling cascades are involved in angiotensin II-induced cellular hypertrophy. The aim of the present study was to determine some of the signaling pathways mediating angiotensin II (Ang II)-induced protein synthesis in vivo in large and small arteries. METHODS: Newly synthesized proteins were labeled during 4 hours with tritiated leucine in conscious control animals, or animals infused for 24 hours with angiotensin II (400 ng/kg/min). Hemodynamic parameters were measure simultaneously. Pharmacological agents affecting signaling cascades were injected 5 hours before the end of Ang II infusion. RESULTS: Angiotensin II nearly doubled the protein synthesis rate in the aorta and small mesenteric arteries, without affecting arterial pressure. The AT1 receptor antagonist Irbesartan antagonized the actions of Ang II. The Ang II-induced protein synthesis was associated with increased extracellular signal-regulated kinases (ERK)1/2 phosphorylation in aortic, but not in mesenteric vessels. Systemic administration of PD98059, an inhibitor of the ERK-1/2 pathway, produced a significant reduction of protein synthesis rate in the aorta, and only a modest decrease in mesenteric arteries. Rapamycin, which influences protein synthesis by alternative signaling, had a significant effect in both vessel types. Rapamycin and PD98059 did not alter basal protein synthesis and had minimal effects on arterial pressure. CONCLUSION: ERK1/2 and rapamycin-sensitive pathways are involved in pressure-independent angiotensin II-induced vascular protein synthesis in vivo. However, their relative contribution may vary depending on the nature of the artery under investigation. PMID- 15134587 TI - Asialoerythropoietin is not effective in the R6/2 line of Huntington's disease mice. AB - BACKGROUND: Huntington's disease (HD) is a hereditary neurodegenerative disorder caused by an expanded CAG repeat in the HD gene. Both excitotoxicity and oxidative stress have been proposed to play important roles in the pathogenesis of HD. Since no effective treatment is available, this study was designed to explore the therapeutic potential of erythropoietin (EPO), a cytokine that has been found to prevent excitotoxicity, and to promote neurogenesis. To avoid the side effects of a raised hematocrit, we used asialoerythropoietin (asialoEPO), a neuroprotective variant of EPO that lacks erythropoietic effects in mice. R6/2 transgenic HD mice were treated with this cytokine from five to twelve weeks of age. RESULTS: We provide new evidence that cell proliferation in the dentate gyrus of the R6/2 hippocampus is reduced by 50% compared to wild-type littermate controls. However, we found that the asialoEPO treatment did not affect the progression of motor symptoms, weight loss or the neuropathological changes. Furthermore, cell proliferation was not enhanced. CONCLUSIONS: We conclude that the chosen protocol of asialoEPO treatment is ineffective in the R6/2 model of HD. We suggest that reduced hippocampal cell proliferation may be an important and novel neuropathological feature in R6 HD mice that could be assessed when evaluating potential therapies. PMID- 15134590 TI - [Quality of nursing documentation in health institutions]. AB - The purpose of this issue is to assess the quality of the nursing documentation adopted by some health structures. The quality of the documentation is considered as a necessary factor to develop an evaluation system of the delivered services. The achieved results point out that all the health structures considered are using a tool to document nursing work, but not every instrument used can be considered complete and effective. PMID- 15134588 TI - Microarray and EST database estimates of mRNA expression levels differ: the protein length versus expression curve for C. elegans. AB - BACKGROUND: Various methods for estimating protein expression levels are known. The level of correlation between these methods is only fair, and systematic biases in each of the methods cannot be ruled out. We here investigate systematic biases in the estimation of gene expression rates from microarray data and from abundance within the Expressed Sequence Tag (EST) database. We suggest that length is a significant factor in biases to measured gene expression rates. As a specific example of the importance of the bias of expression rate with length, we address the following evolutionary question: Does the average C. elegans protein length increase or decrease with expression level? Two different answers to this question have been reported in the literature, one method using expression levels estimated by abundance within the EST database and another using microarrays. We have investigated this issue by constructing the full protein length versus expression curve for C. elegans, using both methods for estimating expression levels. RESULTS: The microarray data show a monotonic decrease of length with expression level, whereas the abundance within the EST database data show a non monotonic behavior. Furthermore, the ratio of the expression level estimated by the EST database to that measured by microarrays is not constant, but rather systematically biased with gene length. CONCLUSIONS: It is suggested that the length bias may lie primarily in the abundance within the EST database method, being not ameliorated by internal standards as it is in the microarray data, and that this bias should be removed before data interpretation. When this is done, both the microarray and the abundance within the EST database give a monotonic decrease of spliced length with expression level, and the correlation between the EST and microarray data becomes larger. We suggest that standard RNA controls be used to normalize for length bias in any method that measures expression. PMID- 15134589 TI - Hypercapnic ventilatory response in mice lacking the 65 kDa isoform of Glutamic Acid Decarboxylase (GAD65). AB - BACKGROUND: Recent reports have shown that there are developmental changes in the ventilatory response to hypercapnia in the rat. These are characterized by an initial large response to carbon dioxide immediately after birth followed by a decline with a trough at one week of age, followed by a return in sensitivity. A second abnormality is seen at postnatal day 5 (P5) rats in that they cannot maintain the increase in frequency for 5 min of hypercapnia. In mice lacking GAD65 the release of GABA during sustained synaptic activation is reduced. We hypothesized that this developmental pattern would be present in the mouse which is also less mature at birth and that GABA mediates this relative respiratory depression. METHODS: In awake C57BL/6J and GAD65-/- mice the ventilatory response to 5% carbon dioxide (CO2) was examined at P2, P4, P6, P7, P12.5, P14.5 and P21.5, using body plethysmography. RESULTS: Minute ventilation (VE) relative to baseline during hypercapnia from P2 through P7 was generally less than from P12.5 onwards, but there was no trough as in the rat. Breaking VE down into its two components showed that tidal volume remained elevated for the 5 min of exposure to 5% CO2. At P6, but not at other ages, respiratory frequency declined with time and at 5 min was less that at 2 and 3 min. GAD65-/- animals at P6 showed a sustained increase in respiratory rate for the five mins exposure to CO2. CONCLUSION: These results show, that in contrast to the rat, mice do not show a decline in minute ventilatory response to CO2 at one week of age. Similar to the rat at P5, mice at P6 are unable to sustain an increase in CO2 induced respiratory frequency and GAD65 contributes to this fall off. PMID- 15134591 TI - [Quality improvement: evolution and use of nursing records in a complex organization: the experience of Policlinico Umberto I of Rome - Italy]. AB - Nursing documentation evolution is the result of an evolution of care and nurses like change of cultural and legislative type that has place, in way more growing, the patient to the center of nursing action. So it affirms that the nursing folder is the tool that allows to record, to plan, to appraise, to document and communicate patient care results, and of not smaller importance, to develop the search implementing nursing assistance turns to qualitative enhancement of the cares. PMID- 15134592 TI - [Descriptive investigation on opinions, knowledge and expectations of hospital nurses of Rome "C" Trust]. AB - In the present article an investigation has been effected to estimate the knowledge, the role and the degree of involvement of the hospital nurses, worked in AUSL Roma C, implies in the clinical experimentations. From results emerged clearly like greater resources must be invested in order to make that the information on the clinical experimentations can circulate in more efficient way and in order to make that it comes recognized to the nurse a role of greater autonomy and competence. PMID- 15134593 TI - [Univerity students' exchange experience]. AB - We report the experience of three University "La Sapienza" of Rome students (two of the Advanced Nursing master science course and one of the 3rd year of Nursing Degree) that participated to the FIPSE programme between European and Transatlantic Universities. The leader of the programme in our Country is Prof J. Sansoni. Web courses have been offered and students participated in a short period exchange at the Capital University with prof Kerr. The experience has been very positive and exciting. We really hope that this programme could be an example to other Italians Colleagues to have a look outside our Country. PMID- 15134594 TI - [Health promoting hospital]. AB - The present report re-assumes the main points of a speech given at the international Congress Health Promoting Hospitals, held in Florence last May and in which the author have represented, through the National Association Consociazione, the Permanent Committee (PCN), co-promotor of the event. The important aspects that have been extrapolated by the nursing point of view specifically concern four priority aspects: the hospital, the patient triad, family and nurse, the professionals as work force and the quality of the Nursing Service given. The Nursing contribution has been remarkable in this Congress very male represented and sustained above all by Hospital and other Health Entities managers. PMID- 15134595 TI - Shared autoimmunity: the time has come. PMID- 15134596 TI - Methotrexate treatment for giant cell arteritis. PMID- 15134598 TI - Henoch-Schonlein purpura. AB - Henoch-Schonlein purpura (HSP) is a multisystem immunoglobulin A-mediated vasculitis with a self-limited course affecting the skin, joints, gastrointestinal tract, and kidneys. HSP occurs most often in children between the ages of 3 and 10 years, and presents classically with a unique distribution of the rash to the lower extremities and the buttocks area. For this reason, a skin biopsy in children is rarely necessary for diagnosis. However, in the very young age children and adults, the rash is not classically distributed, and therefore a skin biopsy is often needed. Although there are currently no prospective controlled studies on the treatment of the different manifestations of HSP, there are several retrospectively designed studies and other physicians' personal experiences supporting the use of steroids in patients with severe gastrointestinal, severe renal, central nervous system, and testicular involvements. Severe renal and central nervous system disease may lead to life threatening conditions, and immunosuppressive agents and plasmapheresis may be needed. The overall prognosis in HSP is excellent, but the long-term morbidity depends on the renal and neurologic involvement. One third to one half of these patients will have one or more recurrences of symptoms, usually within 6 weeks, but may occur as late as 3 to 7 years later. PMID- 15134597 TI - Endothelial cells, antineutrophil cytoplasmic antibodies, and cytokines in the pathogenesis of systemic vasculitis. AB - Endothelial cells play a pivotal role in the pathogenesis of systemic vasculitis. Endothelial cells have significant proinflammatory activities, amplifying and perpetuating the inflammatory process and contributing to vessel regeneration and repair. Significant contributions have improved the understanding of additional ways through which antineutrophil cytoplasmic antibodies (ANCA) may potentiate neutrophil- and monocyte-mediated endothelial cell activation and damage. Signaling pathways mediating ANCA effects have been delineated, and new animal models have demonstrated the pathogenic role of ANCA in the development of systemic vasculitis. Significant efforts have identified anti-endothelial cell antibody specificities and elucidated mechanisms through which these antibodies may promote endothelial cell activation and injury. New ways to assess in vivo endothelial cell damage and dysfunction also have been developed. In addition, besides being a relevant compensatory mechanism for ischemia, angiogenesis may have important proinflammatory functions in vasculitis. The potential relevance of bone-marrow-derived endothelial cell precursors in neovascularization has begun to be appreciated. PMID- 15134600 TI - The role of hypertension in cyclosporine-induced hyperuricemia. PMID- 15134599 TI - Use of imaging studies in the diagnosis of vasculitis. AB - Imaging studies are necessary to determine disease extension and disease activity in the small-vessel vasculitides. Computed tomography (CT) and magnetic resonance imaging (MRI) increase the number of pathologic findings compared with conventional radiography. MRI delineates mucosal inflammation and granulomas in the paranasal sinuses, whereas CT provides information about osseous lesions. CT is superior to MRI for the detection of pulmonary lesions. Radiograph angiography has been the gold standard for medium- and large-vessel vasculitides for decades. Echocardiography and MRI correspond well with conventional angiography to assess cardiac involvement in Kawasaki disease. MRI, CT, and CT angiography are alternative noninvasive techniques to delineate vasculitic lesions in polyarteritis nodosa, Takayasu's arteritis, and large-vessel giant cell arteritis. Duplex ultrasonography has the greatest resolution. It delineates typical artery wall swelling in temporal arteritis and Takayasu's arteritis. Positron emission tomography can assess inflammatory activity of large arteries. PMID- 15134601 TI - Update on identification of pathogenic crystals in joint fluid. AB - Crystal identification in joint fluid has been an essential part of diagnosis of joint disease. Recent advances have included progress in crystal identification on stained slides, attention to aspiration techniques, and arthrocenteses of asymptomatic joints. Challenges remain to increase use and optimize techniques. PMID- 15134603 TI - How crystals damage tissue. AB - Basic calcium phosphate, calcium pyrophosphate dihydrate, and monosodium urate crystals are the most common types of crystals associated with human disease. Although there is a well-established association between these crystals and various forms of joint disease, recent evidence points to an association of basic calcium phosphate crystals with breast cancer and atherosclerosis. Crystal induced tissue damage is affected by degradative proteases, cytokines, chemokines, and prostanoids produced by cells stimulated by crystals. In the case of basic calcium phosphate and calcium pyrophosphate dihydrate crystals, these responses are augmented by the cellular proliferation that results from their induction of mitogenesis. The understanding of the molecular mechanisms involved in generating these pathologic effects has been significantly advanced in recent years. Such advances are essential to the ongoing search for more effective therapies for crystal-associated diseases. PMID- 15134602 TI - Calcium pyrophosphate dihydrate and basic calcium phosphate crystal-induced arthropathies: update on pathogenesis, clinical features, and therapy. AB - Calcium-containing crystals are the most common class for the osteoarthritic joint. They are responsible for acute periarthritis and destructive arthropathies, and for tissue deposits mimicking tumor-like masses. These crystals encompassed mainly calcium pyrophosphate dihydrate and basic calcium phosphate crystals, with the latter being related to hydroxyapatite, carbonate substituted apatite, and octacalcium phosphate. Calcification deposit mechanisms will be reviewed with respect to extracellular inorganic pyrophosphate dysregulation mainly caused by modulation of specific membrane channel disorders. Genetic defects have been extensively studied and identified mutation of specific genes such as ANKH and COL. Pathogenesis of crystal-induced inflammation is related to synovial tissue and direct cartilage activation. Besides classical knee or wrist pseudogout attacks or Milwaukee shoulder arthropathies, clinicians should be aware of other specific common presentations, such as erosive calcifications, spinal cord compression by intraspinal masses, ligamentum flavum calcification, or atypical calcified tophus. Promising clinical results for preventing calcium crystal deposits and cartilage degradation are lacking. Practical imaging tools are needed to monitor reduction of calcification of fibrocartilage and articular cartilage as markers of drug efficacy. PMID- 15134604 TI - Treatment of acute gouty arthritis: one physician's approach and where this management stands relative to developments in the field. AB - An attack of acute gouty arthritis is one the most painful episodes that can be experienced by humans. Fortunately, there are three classes of agents that can effectively terminate the acute attack. These include colchicine, nonsteroidal anti-inflammatory drugs, and glucocorticoids. If therapy with any one of these is initiated promptly after the onset of symptoms, relief should occur quickly. Recent observations have increased the knowledge of crystal-induced acute inflammatory responses. These observations allow a better understanding of the mechanisms of action of these agents and provide the rationale for using them in combination in severe or refractory cases. PMID- 15134607 TI - Perioperative complication rate of total ankle replacement is reduced by surgeon experience. AB - BACKGROUND: Recent studies suggest the perioperative complication rate of total ankle arthroplasty decreases as a surgeon becomes familiar with the procedure. This study tests the hypothesis that the number of perioperative adverse events will decrease as surgeon experience with total ankle replacement increases. METHODS: Ten surgeons completed retrospective chart and radiographic reviews of their first 10 cases as well as 10 subsequent cases of the Scandinavian Total Ankle Replacement (STAR). Not all surgeons completed 10 cases within the allotted time periods, and two patients were excluded for less than 3-month follow-up, resulting in 187 cases for review. The surgeons performed an average of 12.8 (range, 0-61) STARs between these two time periods. Cases were divided into Early Group if they were among the first five STARs a surgeon performed and Late Group if they were after the first five. RESULTS: The average patient age was 60.4 +/- 12.8 years. The etiology of arthrosis included 96 (51%) of 187 posttraumatic, 49 (26%) idiopathic, and 33 (18%) rheumatoid. Patients in Early Group had a 3.1 times greater chance of having a perioperative adverse event (95% CI 1.6-6.1, p <.001), and a 3.2 times greater chance of having a perioperative wound problem (95% CI 1.5-6.8, p =.002) than patients in Late Group. Patients in Early Group took 1 week longer to heal their wounds than patients in Late Group (4.5 vs. 3.5 weeks, p =.046). CONCLUSIONS: This study shows a decrease in the perioperative adverse event rate commensurate with surgeon experience with STAR. In contrast to other reports, this study was unable to show a decrease in the number of perioperative fractures with increasing surgeon experience. This information is important for planning how best to train surgeons new to total ankle replacement and for patient counseling regarding the potential risks of the procedure. PMID- 15134605 TI - Serum uric acid-lowering therapies: where are we heading in management of hyperuricemia and the potential role of uricase. AB - Although allopurinol has been available for approximately 50 years, hyperuricemia and its sequelae are not only prevalent, but the incidence and costs associated with this disorder continue to increase. However, several new therapies have been developed. Recombinant urate oxidase has been useful in the treatment of tumor lysis hyperuricemia, and pegylated urate oxidase shows promise in patients with hyperuricemia and gout. Febuxostat and Y-700 are new oral xanthine oxidase inhibitors that are in human clinical trials. Tailoring of antilipid therapy in selected hyperuricemic and hyperlipidemic patients with fenofibrate may be of benefit in lowering blood cholesterol and uric acid levels. Similarly, treatment of selected hyperuricemic patients who also are hypertensive with losartan or amlodipine may be beneficial in lowering blood pressure and hyperuricemia. Despite these advances, new treatments for hyperuricemia are needed. PMID- 15134608 TI - Extracorporeal shock wave therapy for the treatment of plantar fasciitis. AB - One hundred fifty patients were enrolled in a multicenter, randomized, placebo controlled, prospective, double-blind study to assess the clinical safety and effectiveness of extracorporeal shock wave therapy (ESWT) using the Dornier Epos Ultra for the treatment of plantar fasciitis. The Active Group was treated with electromagnetically generated shocks using ultrasound guidance during a single therapy session. The Control Group received a sham treatment under similar clinical conditions. The groups were demographically similar with respect to age, height, and weight. The average duration of symptoms was nearly 2 years in both groups. All patients were evaluated by the visual analog scale for pain, American Orthopaedic Foot and Ankle Society scores, Roles and Maudsley Score, SF-12 health status questionnaire, and physical examination. The Active Group reported 56% success at 3 months and 94% success at 12 months posttreatment. The Control Group reported 47% success at 3 months posttreatment. Twelve-month data were not collected for the Control Group as they were unblinded at 3 months and offered treatment. ESWT represents a safe treatment option for chronic proximal plantar fasciitis. PMID- 15134609 TI - Bilateral chronic proximal plantar fasciopathy: treatment with electrohydraulic orthotripsy. AB - BACKGROUND: Patients presenting for treatment of chronic plantar fasciopathy often have bilateral involvement. When various nonoperative treatments fail, subsequent intervention may be problematic, especially since bilateral surgery (bilateral fascial release) may not be realistic because of variable, frequently restrictive postoperative weightbearing limitations. METHODS: Twenty-three patients (46 heels) were treated with electrohydraulic high-energy orthotripsy to the plantar entheses of both feet while under the same anesthesia (conscious sedation). Following orthotripsy, all patients immediately were fully weightbearing and resumed normal activities of daily living and work, usually within 24 hours. Progressive return to athletic activities was allowed. Patients were assessed by three outcome parameters: (1) pain measured objectively by a dolorimeter combined with the patient's subjective evaluation of the level of pain; (2) pain after 5 minutes of walking upon arising; and (3) pain with daily activities. All pain measurements were done by the visual analog scale. RESULTS: Patients initially experienced varied pain relief responses. This included earlier pain relief in one heel compared to the other, as well as better pain relief in one heel than the other at the 6- and 12-week evaluations, but with much less variance at the 1-year evaluation. By 3 months following orthotripsy, 28 heels (61%) had good or excellent results. These results were maintained or improved at 1 year. In 18 heels (39%), the outcome was fair or poor. Nineteen heels received a second orthotripsy application; one patient requested a second orthotripsy treatment of only one heel, while nine patients requested a second treatment of both heels. The outcome showed further improvement following the second application of orthotripsy. At 1 year after one or two orthotripsy applications, 19 patients (38 heels) were satisfied with the results in both heels (83%), while four patients (eight heels) still had an unsatisfactory outcome (17%). CONCLUSION: Electrohydraulic high-energy orthotripsy is a reasonable nonincisional method for treating patients with bilateral chronic proximal plantar fasciopathy under a single anesthetic without the prolonged nonweightbearing status often recommended for patients following unilateral open or endoscopic fascial release. PMID- 15134610 TI - Volume of ambulatory care visits and patterns of care for patients diagnosed with plantar fasciitis: a national study of medical doctors. AB - BACKGROUND: Plantar fasciitis is a relatively common disorder of the foot, yet little is known about its prevalence, what types of physicians see patients with the disorder, or how, on a national scale, patients are typically managed. The purpose of this study was to generate national estimates of the volume of patient visits and characteristics of care given to patients diagnosed with plantar fasciitis by medical doctors. METHODS: Data from the National Ambulatory Medical Care Survey (NAMCS) and the National Hospital Ambulatory Medical Care Survey (NHAMCS) for the years 1995-2000 were studied. These are multistage probability sample surveys of visits to office-based physicians (NAMCS) and visits to nonfederal, short-stay, and general hospitals (NHAMCS) to consult doctors of medicine and doctors of osteopathy. Sample data have been weighted to produce national estimates. Data describing the number of patient visits for plantar fasciitis and the characteristics of the care given during those visits were summarized using univariate analyses. Data were combined for the 6-year period to increase the reliability of the estimates, and figures are presented as annual averages. RESULTS: Approximately 1 million patient visits per year were made to office-based physicians and hospital outpatient departments for the diagnosis and treatment of plantar fasciitis during 1995-2000. Approximately 62% of all visits were made to primary care practitioners, and 31% were made to orthopaedic surgeons. Patient visits for plantar fasciitis accounted for approximately 1% of all patient visits to orthopedic surgeons. Pain medication, including nonsteroidal anti-inflammatory drugs (NSAIDs), was the most frequently used intervention (47% of visits). Exercise counseling was cited at 26% of visits, and physical therapy was ordered or provided at 19% of visits. CONCLUSIONS: This research suggests that plantar fasciitis is a relatively common disorder that is seen by several physician specialties. The disorder is not managed in a consistent way. Rather, there appears to be a large amount of variation in the way that these patients are managed. These findings support the argument that additional research is needed to identify effective interventions for plantar fasciitis and to determine if physician specialty influences treatment outcome. PMID- 15134611 TI - Impact of demographic and impairment-related variables on disability associated with plantar fasciitis. AB - BACKGROUND: Plantar fasciitis is a common foot disorder that impacts many functional activities. Research that quantifies the impact that plantar fasciitis has on function is lacking. In addition, little is known about which variables are associated with disability in patients with plantar fasciitis. The first purpose of this study was to determine if age, gender, body mass index, pain intensity, chronicity of symptoms, or ankle dorsiflexion range of motion was associated with disability in patients with plantar fasciitis. The second purpose was to describe the impact that plantar fasciitis has on functional status in the context of five functional domains: household activities of daily living, usual work and hobbies, nonweightbearing activities, walking-related activities, and running-related activities. METHODS: Fifty consecutive patients diagnosed with unilateral plantar fasciitis were recruited. Demographic and impairment data were collected and all patients completed the Lower Extremity Functional Scale (LEFS), a validated self-report measure of disability. Multiple regression analysis was used to describe the association between the variables and disability. Graphs depicting five domains of function derived from the LEFS were generated to describe the extent of disability. RESULTS: Body mass index (BMI) was the only variable that was significantly associated with disability (F = 9.87, p =.003). Measures of pain intensity, ankle dorsiflexion, age, gender, chronicity, and time spent weightbearing were not related to disability. Plantar fasciitis showed distinct patterns of disability depending on the functional domain that was assessed. CONCLUSIONS: With the exception of BMI, impairment and demographic variables do not predict the extent of functional loss in patients with plantar fasciitis. The most likely domains of function to be at least moderately affected in patients with plantar fasciitis are running-related activities and usual work or hobbies. PMID- 15134612 TI - Fibular position in relation to lateral ankle instability. AB - BACKGROUND: The position of the fibula within the ankle mortise may be a factor contributing to recurrent ankle instability. The current study was performed to determine whether significant differences in fibular position exist in a population of patients who underwent lateral ankle stabilization procedures. The hypothesis that a fibula positioned posteriorly within the mortise predisposes the ankle to chronic instability was evaluated. METHODS: Sixty-five CT/MRI scans of patients who underwent lateral ankle stabilization procedures from 1998 to 2001 were reviewed. The position of the fibula in relation to the tibia at the ankle mortise was expressed as the axial malleolar index (AMI). A greater AMI corresponds to a more posterior fibula. The AMI was also calculated from 65 CT/MRI scans performed on control patients who had no ankle instability. RESULTS: The average AMI in the study group was 17 degrees +/- 6 degrees (SD) compared with an average of 9 degrees +/- 4 degrees (SD) in the control group (p <.01). Therefore, the fibula was nearly twice as posterior in patients undergoing ankle reconstruction. In 42/65 (65%) study patients, the AMI was greater than 15 degrees. Only 5/65 (8%) control patients had AMI values greater than 15 degrees. This corresponds to an eightfold greater likelihood of AMI greater that 15 degrees in the instability group than in the control group. CONCLUSIONS: This study supports the hypothesis that a posteriorly positioned fibula predisposes to ankle instability. PMID- 15134613 TI - Multiple accessory peroneal muscles: a cause of chronic lateral ankle pain. PMID- 15134614 TI - Distal lower extremity sarcomas: frequency of occurrence and patient survival rate. AB - BACKGROUND: Primary sarcomas in the distal leg, tibia, fibula, ankle, and foot are uncommon and are believed to be less malignant than those that arise in other sites, but only limited information is available to support this contention. METHODS: Using a computerized system containing extensive information regarding over 14,000 patients, mostly with tumors treated by our center over a 25-year period, 175 sarcomatous lesions with MSTS stage I, II, and III were located in the distal lower extremity. These were compared with 2367 lesions of similar diagnoses in other body parts. The principal studies included diagnostic distribution and outcome (recorded as death as a result of disease). Data were compared for diagnosis, gender, age, Musculoskeletal Tumor Society (MSTS) stage, anatomic site, and treatment methods and evaluated statistically by chi-square methods. RESULTS: The most frequent distal lower limb tumors were synovial cell sarcoma, osteosarcoma, and Ewing's tumor and the percentage distribution of the various tumors for that site as compared with the rest of the body was quite different. Of even more importance was the remarkable difference in outcome with the death rate for the lower limb tumors set at 10%, while the same tumors at other sites had a death rate of 27% (p <.000002). Furthermore, gender, stage, age, and the type of operative procedure showed highly significant differences between the tumors of the lower leg and those of the remainder of the body. CONCLUSIONS: On the basis of these data, it is evident that far fewer sarcomas occur in the lower leg, tibia, fibula, ankle, and foot than in other body sites and that their diagnostic distribution is different. Of perhaps greater interest is the fact that the rate of metastasis and death is markedly reduced for this site as compared with others. The authors speculate on the causes of this remarkable alteration in numbers and outcome. PMID- 15134615 TI - Infectious and healing complications after elective orthopaedic foot and ankle surgery during tumor necrosis factor-alpha inhibition therapy. AB - BACKGROUND: Biologic response modifiers are assuming a larger role in the management of patients with rheumatoid arthritis. The tumor necrosis factor-alpha (TNF-alpha) inhibitors etanercept and infliximab improve patient symptoms and function. However, these agents have been associated with a risk for healing and infectious complications due to systemic blockade of TNF-alpha, a ubiquitous mediator required in the normal inflammatory response in tissue healing and infection surveillance. This study analyzed the risk of healing/infectious complications in patients undergoing elective foot and ankle surgery while being treated with TNF-alpha inhibitors etanercept and infliximab. METHODS: Patients with rheumatoid arthritis undergoing elective foot and ankle surgery over a 12 month period were prospectively followed for the development of complications in the postoperative period. All patients continued their antirheumatic medication schedule unaltered in the perioperative period. Data collected included sex, age, all medications used to treat rheumatoid arthritis, smoking history, and number of orthopaedic foot and ankle procedures performed. Patients were then stratified into two groups based on the use of immunomodulation via TNF-alpha inhibition (group 1) versus patients who did not receive TNF-alpha inhibition therapy (group 2). Groups 1 and 2 were followed and compared for the development of infectious/healing complications. RESULTS: Thirty-one patients were enrolled in the study. Group 1 (n = 16) and group 2 (n = 15) patients were comparable for sex distribution, number of orthopaedic procedures performed, and use of steroids, methotrexate, leflunamide, and nonsteroidal anti-inflammatory drugs. Group 1 contained six times the number of smokers in group 2. At mean follow-up of 10.6 months (group 1) and 9.7 months (group 2), healing or infectious complications were similar in both groups. However, when total complications (healing + infection) were analyzed, group 1 (TNF-alpha inhibition, "higher risk") patients demonstrated a lower complication rate (p =.033). CONCLUSIONS: The data suggest that in patients with rheumatoid arthritis undergoing elective foot and ankle surgery, the use of TNF-alpha inhibition agents may be safely undertaken in the perioperative period without increasing the risk of healing or infectious complications. PMID- 15134616 TI - Rheumatoid necroses in the forefoot. AB - BACKGROUND: The incidence and morphological features of rheumatoid necroses (RN) in the forefoot are unknown. METHODS: Biopsy material from 223 patients with seropositive rheumatoid arthritis (RA) was examined morphologically. The incidence and morphological features of RN in the forefoot were evaluated. RESULTS: RN was found in 65% of cases, which had not been suspected clinically. It was correlated with pain in 69% of cases. CONCLUSIONS: Rheumatoid necrosis contributes to the development of pain and foot deformation. It can be a significant factor worsening the clinical behavior in seropositive RA patients. PMID- 15134617 TI - Chopart joint fracture-dislocation: initial open reduction provides better outcome than closed reduction. AB - Injury cause, treatment, and long-term results [American Orthopaedic Foot and Ankle Society (AOFAS) Midfoot Score, Hannover Scoring System, Hannover Outcome Questionnaire] of patients with Chopart joint dislocations or fracture dislocations were evaluated. Between 1972 and 1997, 100 patients with 110 Chopart joint dislocations were treated in the authors' institution. Pure Chopart joint dislocations were observed in 28 (25%) feet, fracture-dislocations in 60 (55%) feet, and combined Chopart-Lisfranc joint fracture-dislocations in 22 (20%) feet. The primary treatment was operative in 91 (83%) feet and nonoperative in 19 (17%) feet. Sixty-five (65%) patients had follow-up after an average of 9 years (range, 2-25 years). The mean scores of the entire follow-up group were: AOFAS score, 75 points; Hannover Scoring System, 69 points (maximium possible score = 100 points); Hannover Outcome Questionnaire, 68 points (maximium possible score = 100 points). There were no differences between the scores for pure dislocations or fracture-dislocations of the Chopart joint, but significantly lower scores were noted with combined Chopart-Lisfranc joint fracture-dislocations. In all three injury pattern groups, an initial anatomic reduction was essential for good results. The high functional restrictions in Chopart dislocations can most likely be minimized with initial open reduction, especially in fracture-dislocations. A closed reduction yielded good results only with pure dislocations, when anatomic conditions could be restored, or if there were contraindications to surgery. PMID- 15134618 TI - Calcaneocuboid joint instability: a novel operative technique for anatomic reconstruction. AB - A case history of a 13-year-old female national top-level gymnast, suffering from calcaneocuboid joint instability, is presented. The procedure was done as an anatomic repair by capsular reefing, which was augmented using a local periosteal flap. Initially, the athlete twisted her ankle. Clinical investigation revealed no sign of a lateral ankle ligament injury, but following this initial examination, recurrent giving-way of the foot occurred. She additionally felt significant but diffuse pain on the lateral side of the foot during loading in training and competition. For 2 months she was unable to run and conservative treatment failed. Diagnosis of a calcaneocuboid instability was established 4 months after the initial lesion by clinical and x-ray stress examination of the calcaneocuboid joint. Open surgery was successfully performed. Early functional posttreatment was done and the patient returned to full high-level gymnastics ability 16 weeks after surgery. Two years later, a similar injury occurred to the opposite calcaneocuboid joint and the same operative procedure again led to full sports ability. PMID- 15134619 TI - Ankle and subtalar kinematics measured with intracortical pins during the stance phase of walking. AB - BACKGROUND: The absence of external landmarks on the talus has rendered the description of ankle and subtalar joint kinematics difficult. Abnormal motion at these joints has, however, been implied in the etiology of an array of lower extremity overuse injuries. METHODS: Intracortical pins were inserted under local anesthesia in the tibia, talus, and calcaneus with external marker clusters traced by a video motion analysis system. Kinematic data were collected during walking trials on a flat surface for three subjects. Gait pattern was monitored by comparison of ground reaction force curves during stance phase with and without the pins inserted. RESULTS: Results were presented in terms of helical axis orientation for both joints and the component rotations about these axes. Large intersubject differences were seen in both ankle and subtalar joint helical axis orientation. Maximum rotations over the complete stance phase for the ankle and subtalar joints respectively were: eversion/inversion, 6.3 degrees and 8.3 degrees; dorsiflexion/plantarflexion, 18.7 degrees and 3.7 degrees; and abduction/adduction, 5.0 degrees and 6.1 degrees. CONCLUSIONS: The majority of ankle eversion/inversion occurred at the subtalar joint; however, the ankle component cannot be ignored. Abduction/adduction range of motion at the subtalar joint was surprisingly high, indicating that this component motion during walking is not purely attributable to the ankle joint. Future research should include greater subject numbers in order to present more universally applicable results. CLINICAL RELEVANCE: The in vivo kinematics of the talus during weightbearing activity are poorly understood. The description of this motion may assist in the structuring of clinical rehabilitation and in the design and insertion of ankle joint prostheses. PMID- 15134620 TI - Stabilization of Lisfranc joint injuries: a biomechanical study. AB - BACKGROUND: Lisfranc joint injuries are often misdiagnosed, leading to significant morbidity. Methods for anatomic reduction of the tarsometatarsal joint include closed reduction with casting or surgical stabilization with either Kirschner wires and/or cortical screw fixation. Controversy exists as to which fixation technique offers optimal stability. In the present study, the biomechanical stability of three fixation methods was tested: (1) four Kirschner wires, (2) three cortical screws plus two Kirschner wires, and (3) five cortical screws. METHODS: Ten matched pairs of fresh-frozen cadaveric feet were dissected to their ligamentous and capsular elements. The tarsometatarsal ligaments were completely transected to replicate a Lisfranc dislocation; the "injury" was reduced and stabilized using one of the three methods. Biomechanical studies were performed by applying a 100-N cyclic load physiologically distributed to the plantar aspect of the metatarsal heads. Displacement and force measurements were taken from the first and fifth metatarsal heads. Average stiffness of each construct was calculated from the force displacement curves. RESULTS AND CONCLUSIONS: Method 2 provided significantly more stability than Kirschner wire fixation. Method 3 created more stiffness than method 2 at the medial portion of the foot; no statistical difference between the two methods was evident at the lateral foot. CLINICAL RELEVANCE: Cortical screw fixation provides a more rigid and stable method of fixation for Lisfranc injuries as compared to Kirschner wire fixation. This fixation method allows maintenance of anatomic reduction and possibly earlier mobilization with a decreased risk of posttraumatic arthrosis. PMID- 15134621 TI - Pseudo os trigonum sign: missed posteromedial talar facet fracture, Giuffrida, AY, et al., Foot Ankle Int. 24(8):642-649, 2003. PMID- 15134622 TI - Interactions of the allogeneic effector leukemic T cell line, TALL-104, with human malignant brain tumors. AB - TALL-104 is a human leukemic T cell line that expresses markers characteristic of both cytotoxic T lymphocytes and natural killer cells. TALL-104 cells are potent tumor killers, and the use of lethally irradiated TALL-104 as cellular therapy for a variety of tumors has been explored. We investigated the interactions of TALL-104 cells with human brain tumor cells. TALL-104 cells mediated increased lysis of a panel of brain tumor cells at low effector-to-target ratios over time. We obtained evidence that TALL-104 cells injured glioma cells by both apoptotic and necrotic pathways. A 7-amino actinomycin D flow cytometry assay revealed that the percentages of both apoptotic and necrotic glioma cells increased after TALL 104 cell/glioma cell coincubations. Fluorescent microscopy studies and a quantitative morphologic assay confirmed that TALL-104 cell/glioma cell interactions resulted in tumor cell apoptosis. Cytokines are secreted when TALL 104 cells are coincubated with brain tumor cells; however, morphologic analysis assays revealed that the soluble factors contained within clarified supernates obtained from 4 h coincubates added back to brain tumor cell cultures did not trigger the glioma apoptosis. TALL-104 cells do not express Fas ligand, even upon coincubation with glioma targets, which suggests that the Fas/Fas ligand apoptotic pathway is not likely responsible for the cell injury observed. We obtained evidence that cell injury is calcium dependent and that lytic granule exocytosis is triggered by contact of TALL-104 cells with human glioma cells, suggesting that this pathway mediates glioma cell apoptosis and necrosis. PMID- 15134623 TI - Analysis of 1p, 19q, 9p, and 10q as prognostic markers for high-grade astrocytomas using fluorescence in situ hybridization on tissue microarrays from Radiation Therapy Oncology Group trials. AB - Survival periods vary considerably for patients with high-grade astrocytomas, and reliable prognostic markers are not currently available. We therefore investigated whether genetic losses from chromosomes 1p, 19q, 9p, or 10q were associated with survival in 89 high-grade astrocytomas using tissue microarrays (TMAs) derived from Radiation Therapy Oncology Group clinical trials. Cases included 15 anaplastic astrocytomas (AAs) and 74 glioblastomas (GBMs) selected on the basis of survival times significantly shorter or longer than the expected median. Genetic analysis was performed by TMA-fluorescence in situ hybridization (FISH) on array sections using 8 DNA probes, including those directed at 1p32, 19q13.4, 9p21 (p16/CDKN2A), and 10q (PTEN and DMBT1). Genetic status for each locus was correlated with patient survival group, and data were analyzed by using Fisher's exact test of association (adjusted P = 0.025). Losses of chromosome 1p, either alone or in combination with 19q, were encountered in only 2 cases, both AAs. This contrasts with oligodendrogliomas, in which combined 1p and 19q losses are frequent and predictive of prolonged survival. Solitary 19q loss was noted in 3/15 AAs and in 7/70 GBMs and was more frequent in the long-term survival group (P = 0.041, AA and GBM combined). Chromosome 9p loss was seen in 5/8 AAs and 39/57 GBMs, whereas chromosome 10q loss was detected in 4/15 AAs and 48/68 GBMs. The 9p and 10q deletions were slightly more frequent in short-term survivors, though none of the comparisons achieved statistical significance. Long-term and short-term survival groups of high-grade astrocytomas appear to have dissimilar frequencies of 19q, 9p, and 10q deletions. TMA-FISH is a rapid and efficient way of evaluating genetic alterations in such tumors. PMID- 15134624 TI - Comparative pharmacokinetics of 14C-sucrose in RG-2 rat gliomas after intravenous and convection-enhanced delivery. AB - We compared tissue and plasma pharmacokinetics of 14C-sucrose in subcutaneous RG 2 rat gliomas after administration by 3 routes, intravenous bolus (i.v.-B; 50 microCi over 30 s), continuous i.v. infusion (i.v.-C, 50 microCi at a constant rate), and convection-enhanced delivery (CED, 5 microCi infused at a rate of 0.5 microl/min), and for 3 experimental durations, 0.5, 2, and 4 h. Plasma, tumor, and other tissue samples were obtained to measure tissue radioactivity. Plasma radioactivity in the CED group increased exponentially and lagged only slightly behind the IV-C group. After 90 min, plasma values were similar in all. Mean tumor radioactivity was 100 to 500 times higher in the CED group at each time point than in the i.v.-B and i.v.-C groups. Tumor radioactivity was homogeneous in the i.v. groups at 0.5 h and inhomogeneous at 1 and 2 h. In CED, radioactivity distribution was inhomogeneous at all 3 time points; highest concentrations were in tissue around tumor and in necrosis, while viable tumor contained the lowest and sometimes negligible amounts of isotope. Systemic tissue radioactivity values were similar in all groups. Efflux of 14C-sucrose from tumors was evaluated in intracerebral tumors (at 0.5, 1, 2, and 4 h) and subcutaneous tumors (at 0 to 0.5 h). Less than 5% of 14C activity remained in intracerebral tumors at each time point. The efflux half-time from the subcutaneous tumors was 7.3 +/- 0.7 min. These results indicate rapid efflux of drug from brain tumor and marked heterogeneity of drug distribution within tumor after CED administration, both of which may be potentially limiting factors in drug delivery by this method. PMID- 15134626 TI - Permanent iodine 125 brachytherapy in patients with progressive or recurrent glioblastoma multiforme. AB - This study reports the initial experience at the University of California San Francisco (UCSF) with tumor resection and permanent, low-activity iodine 125 (125I) brachytherapy in patients with progressive or recurrent glioblastoma multiforme (GM) and compares these results to those of similar patients treated previously at UCSF with temporary brachytherapy without tumor resection. Thirty eight patients with progressive or recurrent GM were treated at UCSF with repeat craniotomy, tumor resection, and permanent, low-activity 125I brachytherapy between June 1997 and May 1998. Selection criteria were Karnofsky performance score > or =60, unifocal, contrast-enhancing, well-circumscribed progressive or recurrent GM that was judged to be completely resectable, and no evidence of leptomeningeal or subependymal spread. The median brachytherapy dose 5 mm exterior to the resection cavity was 300 Gy (range, 150-500 Gy). One patient was excluded from analysis. Median survival was 52 weeks from the date of brachytherapy. Age, Karnofsky performance score, and preimplant tumor volume were all statistically significant on univariate analyses. Multivariate analysis for survival showed only age to be significant. Median time to progression was 16 weeks. Both univariate and multivariate analysis of freedom from progression showed only preoperative tumor volume to be significant. Comparison to temporary brachytherapy patients showed no apparent difference in survival time. Chronic steroid requirements were low in patients with minimal postoperative residual tumor. We conclude that permanent 125I brachytherapy for recurrent or progressive GM is well tolerated. Survival time was comparable to that of a similar group of patients treated with temporary brachytherapy. PMID- 15134625 TI - Endocrine outcome in children with medulloblastoma treated with 18 Gy of craniospinal radiation therapy. AB - Craniospinal radiation therapy (CSRT) combined with chemotherapy results in significant endocrine morbidity. Between 1987 and 1990, a trial using 18 Gy was conducted to treat 10 young children with medulloblastoma. There were 7 survivors. We compared the endocrine outcome in these children (group 18 Gy) to that of a comparable group treated with conventional doses of CSRT that ranged from 23 to 39 Gy (group CD). Both groups had an identical history of chemotherapy and tumor stage and were treated with recombinant growth hormone therapy (rhGH). The mean age of group 18 Gy at diagnosis was 4.0 years, and rhGH treatment was initiated in 6 children at age 9.2 years. Group CD (12 children) was diagnosed at a mean age of 5.8 years and rhGH started in 11 children at a mean age of 9.6 years. The dose of rhGH used in both groups was identical (0.3 mg/kg/wk). For group 18 Gy, adult heights and sitting heights (a mean standard deviation score of -1.01 +/- 1.11 and -1.62 +/- 1.16, respectively) were statistically greater (P < 0.05) than those for group CD (mean standard deviation score of -2.04 +/- 0.83 and -3.16 +/- 1.43, respectively). Moreover, adult heights of group 18 Gy were not different from midparental heights, unlike group CD, whose adult heights were less than midparental heights (P < 0.0001). Of other endocrine sequelae, 10 patients of the CD group were hypothyroid, 3 had adrenal insufficiency, 3 had hypogonadism, and 2 had early puberty. In contrast, within group 18 Gy, only 1 was hypothyroid (P = 0.006) and 1 had early puberty. We conclude that endocrine morbidity was significantly reduced with 18 Gy CSRT in young children with medulloblastoma. PMID- 15134627 TI - Refining the staging evaluation of pineal region germinoma using neuroendoscopy and the presence of preoperative diabetes insipidus. AB - Treatment strategies for CNS germinoma are currently evolving. Current approaches include reducing the volume and dose of radiation by adding pre-irradiation chemotherapy. Very accurate staging is necessary with such an approach to prevent failures. Eight consecutive patients with pineal germinoma at one institution underwent endoscopic surgery for tumor biopsy, direct visualization of the third ventricular region, and third ventriculostomy for those with hydrocephalus. All patients were treated with 4 cycles of chemotherapy. Conformal field radiation therapy followed, with the dose to the tumor bed dependent on the response to chemotherapy. Patients who had MRI, endoscopic, or cerebrospinal fluid evidence of multicentric or disseminated disease also received craniospinal radiation. Six patients had diabetes insipidus (DI) at presentation. All 6 had tumor studding the floor of the third ventricle on endoscopic visualization, while only 4 of those patients had MRI evidence of disease in that region. All patients have completed therapy and are alive, with no evidence of disease at median follow-up of 31.5 months from diagnosis. Direct endoscopic visualization of the third ventricular region may be more sensitive than MRI for evaluating the presence of suprasellar disease and appears to add important information. This parameter should be added to the staging evaluation when feasible. In this series, the presence of DI was 100% predictive of suprasellar disease, even when the MRI was negative for involvement of that region. Patients should be evaluated for DI as part of the initial staging, and if it is present, the patients should be treated for suprasellar disease regardless of MRI findings. PMID- 15134628 TI - Phase 2 trial of BCNU plus irinotecan in adults with malignant glioma. AB - In preclinical studies, BCNU, or 1,3-bis(2-chloroethyl)-1-nitrosourea, plus CPT 11 (irinotecan) exhibits schedule-dependent, synergistic activity against malignant glioma (MG). We previously established the maximum tolerated dose of CPT-11 when administered for 4 consecutive weeks in combination with BCNU administered on the first day of each 6-week cycle. We now report a phase 2 trial of BCNU plus CPT-11 for patients with MG. In the current study, BCNU (100 mg/m2) was administered on day 1 of each 6-week cycle. CPT-11 was administered on days 1, 8, 15, and 22 at 225 mg/m2 for patients receiving CYP3A1- or CYP3A4-inducing anticonvulsants and at 125 mg/m2 for those not on these medications. Newly diagnosed patients received up to 3 cycles before radiotherapy, while recurrent patients received up to 8 cycles. The primary end point of this study was radiographic response, while time to progression and overall survival were also assessed. Seventy-six patients were treated, including 37 with newly diagnosed tumors and 39 with recurrent disease. Fifty-six had glioblastoma multiforme, 18 had anaplastic astrocytoma, and 2 had anaplastic oligodendroglioma. Toxicities (grade > or =3) included infections (13%), thromboses (12%), diarrhea (10%), and neutropenia (7%). Interstitial pneumonitis developed in 4 patients. Five newly diagnosed patients (14%; 95% CI, 5%-29%) achieved a radiographic response (1 complete response and 4 partial responses). Five patients with recurrent MG also achieved a response (1 complete response and 4 partial responses; 13%; 95% CI, 4% 27%). More than 40% of both newly diagnosed and recurrent patients achieved stable disease. Median time to progression was 11.3 weeks for recurrent glioblastoma multiforme patients and 16.9 weeks for recurrent anaplastic astrocytoma/ anaplastic oligodendroglioma patients. We conclude that the activity of BCNU plus CPT-11 for patients with MG appears comparable to that of CPT-11 alone and may be more toxic. PMID- 15134629 TI - Phase 1 trial of irinotecan plus BCNU in patients with progressive or recurrent malignant glioma. AB - Irinotecan is a topoisomerase I inhibitor previously shown to be active in the treatment of malignant glioma. We now report the results of a phase 1 trial of irinotecan plus BCNU, or 1,3-bis(2-chloroethyl)-1-nitrosourea, for patients with recurrent or progressive MG. Irinotecan dose escalation occurred independently within 2 strata: patients receiving enzyme-inducing antiepileptic drugs (EIAEDs) and patients not receiving EIAEDs. BCNU was administered at a dose of 100 mg/m2 over 1 h every 6 weeks on the same day as the first irinotecan dose was administered. Irinotecan was administered intravenously over 90 min once weekly. Treatment cycles consisted of 4 weekly administrations of irinotecan followed by a 2-week rest with dose escalation in cohorts of 3 to 6 patients. Seventy-three patients were treated, including 49 patients who were on EIAEDs and 24 who were not on EIAEDs. The maximum tolerated dose for patients not on EIAEDs was 125 mg/m2. The maximum tolerated dose for patients on EIAEDs was 225 mg/m2. Dose limiting toxicity was evenly distributed among the following organ systems: pulmonary, gastrointestinal, cardiovascular, neurologic, infectious, and hematologic, without a clear predominance of toxicity involving any one organ system. There was no evidence of increasing incidence of toxicity involving one organ system as irinotecan dose was escalated. On the basis of these results, we conclude that the recommended doses of irinotecan for a phase 2 clinical trial when given in combination with BCNU (100 mg/m2) are 225 mg/m2 for patients on EIAEDs and 125 mg/m2 for patients not on EIAEDs. PMID- 15134630 TI - Brain metastases in melanoma: roles of neurotrophins. AB - Brain metastasis, which occurs in 20% to 40% of all cancer patients, is an important cause of neoplastic morbidity and mortality. Successful invasion into the brain by tumor cells must include attachment to microvessel endothelial cells, penetration through the blood-brain barrier, and, of relevance, a response to brain survival and growth factors. Neurotrophins (NTs) are important in brain invasive steps. Human melanoma cell lines express low-affinity NT receptor p75NTR in relation to their brain-metastatic propensity with their invasive properties being regulated by NGF, or nerve growth factor, the prototypic NT. They also express functional TrkC, the putative receptor for the invasion-promoting NT-3. In brain-metastatic melanoma cells, NTs promote invasion by enhancing the production of extracellular matrix (ECM)-degradative enzymes such as heparanase, an enzyme capable of locally destroying both ECM and the basement membrane of the blood-brain barrier. Heparanase is an endo-beta-d-glucuronidase that cleaves heparan sulfate (HS) chains of ECM HS proteoglycans, and it is a unique metastatic determinant because it is the dominant mammalian HS degradative enzyme. Brain-metastatic melanoma cells also produce autocrine/paracrine factors that influence their growth, invasion, and survival in the brain. Synthesis of these factors may serve to regulate NT production by brain cells adjacent to the neoplastic invasion front, such as astrocytes. Increased NT levels have been observed in tumor-adjacent tissues at the invasion front of human brain melanoma. Additionally, astrocytes may contribute to the brain-metastatic specificity of melanoma cells by producing NT-regulated heparanase. Trophic, autocrine, and paracrine growth factors may therefore determine whether metastatic cells can successfully invade, colonize, and grow in the CNS. PMID- 15134631 TI - The dawn of high-resolution structure for the queen of ion channels. PMID- 15134632 TI - The spatial code for odors is changed by conditioning. PMID- 15134633 TI - Semaphorin signaling unplugged; a nervy AKAP cAMP(s) out on plexin. PMID- 15134634 TI - Differential functions of the C. elegans FGF receptor in axon outgrowth and maintenance of axon position. AB - Wiring of the nervous system requires that axons navigate to their targets and maintain their correct positions in axon fascicles after termination of axon outgrowth. We show here that the C. elegans fibroblast growth factor receptor (FGFR), EGL-15, affects both processes in fundamentally distinct manners. FGF dependent activation of the EGL-15 tyrosine kinase and subsequently the GTPase LET-60/ras is required within epidermal cells, the substratum for most outgrowing axon, for appropriate outgrowth of specific axon classes to their target area. In contrast, genetic elimination of the FGFR isoform EGL-15(5A), defined by the inclusion of an alternative extracellular interimmunoglobulin domain, has no consequence for axon outgrowth but leads to a failure to postembryonically maintain axon position within defined axon fascicles. An engineered, secreted form of EGL-15(5A) containing only its ectodomain is sufficient for maintenance of axon position, thus providing novel insights into receptor tyrosine kinase function and the process of maintaining axon position. PMID- 15134635 TI - Genetic identification of spinal interneurons that coordinate left-right locomotor activity necessary for walking movements. AB - The sequential stepping of left and right limbs is a fundamental motor behavior that underlies walking movements. This relatively simple locomotor behavior is generated by the rhythmic activity of motor neurons under the control of spinal neural networks known as central pattern generators (CPGs) that comprise multiple interneuron cell types. Little, however, is known about the identity and contribution of defined interneuronal populations to mammalian locomotor behaviors. We show a discrete subset of commissural spinal interneurons, whose fate is controlled by the activity of the homeobox gene Dbx1, has a critical role in controlling the left-right alternation of motor neurons innervating hindlimb muscles. Dbx1 mutant mice lacking these ventral interneurons exhibit an increased incidence of cobursting between left and right flexor/extensor motor neurons during drug-induced locomotion. Together, these findings identify Dbx1-dependent interneurons as key components of the spinal locomotor circuits that control stepping movements in mammals. PMID- 15134636 TI - Structural analysis of the voltage-dependent calcium channel beta subunit functional core and its complex with the alpha 1 interaction domain. AB - Voltage-dependent calcium channels (VDCC) are multiprotein assemblies that regulate the entry of extracellular calcium into electrically excitable cells and serve as signal transduction centers. The alpha1 subunit forms the membrane pore while the intracellular beta subunit is responsible for trafficking of the channel to the plasma membrane and modulation of its electrophysiological properties. Crystallographic analyses of a beta subunit functional core alone and in complex with a alpha1 interaction domain (AID) peptide, the primary binding site of beta to the alpha1 subunit, reveal that beta represents a novel member of the MAGUK protein family. The findings illustrate how the guanylate kinase fold has been fashioned into a protein-protein interaction module by alteration of one of its substrate sites. Combined results indicate that the AID peptide undergoes a helical transition in binding to beta. We outline the mechanistic implications for understanding the beta subunit's broad regulatory role of the VDCC, particularly via the AID. PMID- 15134637 TI - Subunit configuration of heteromeric cone cyclic nucleotide-gated channels. AB - Cone photoreceptor cyclic nucleotide-gated (CNG) channels are thought to be tetrameric assemblies of CNGB3 (B3) and CNGA3 (A3) subunits. We have used functional and biochemical approaches to investigate the stoichiometry and arrangement of these subunits in recombinant channels. First, tandem dimers of linked subunits were used to constrain the order of CNGB3 and CNGA3 subunits; the properties of channels formed by B3/B3+A3/A3 dimers, or A3/B3+B3/A3 dimers, closely resembled those of channels arising from B3+A3 monomers. Functional markers in B3/B3 (or A3/A3) dimers confirmed that both B3 subunits (and both A3 subunits) gained membership into the pore-forming tetramer and that like subunits were positioned adjacent to each other. Second, chemical crosslinking and co immunoprecipitation studies using epitope-tagged monomer subunits both demonstrated the presence of two CNGB3 subunits in cone channels. Together, these data support a preferred subunit arrangement for cone CNG channels (B3-B3-A3-A3) that is distinct from the 3A:1B configuration of rod channels. PMID- 15134638 TI - Stoichiometry and assembly of olfactory cyclic nucleotide-gated channels. AB - Native ion channels are precisely tuned to their physiological role in neuronal signaling. This tuning frequently involves the controlled assembly of heteromeric channels comprising multiple types of subunits. Cyclic nucleotide-gated (CNG) channels of olfactory neurons are tetramers and require three types of subunits, CNGA2, CNGA4, and CNGB1b, to exhibit properties necessary for olfactory transduction. Using fluorescently tagged subunits and fluorescence resonance energy transfer (FRET), we find the subunit composition of heteromeric olfactory channels in the surface membrane is fixed, with 2:1:1 CNGA2:CNGA4:CNGB1b. Furthermore, when expressed individually with CNGA2, CNGA4 and CNGB1b subunits were still present in only a single copy and, when expressed alone, did not self assemble. These results suggest that the precise assembly of heteromeric olfactory channels results from a mechanism where CNGA4 and CNGB1b subunits have a high affinity for CNGA2 but not for self-assembly, precluding more than one CNGA4 or CNGB1b subunit in the channel complex. PMID- 15134639 TI - Plastic elimination of functional glutamate release sites by depolarization. AB - To examine persisting effects of depolarizing rises in extracellular potassium concentration ([K+](o)) on synapses, we depolarized cells to simulate ischemia like rises in [K+](o). Elevated [K+](o) for 1-16 hr severely depressed glutamate signaling, while mildly depressing GABA transmission. The glutamate-specific changes were plastic over several hours and involved a decrease in the size of the pool of releasable vesicles. Rather than a reduction of the number of vesicles per release site, the change involved functional elimination of release sites. This change was clearly dissociable from a second effect, depressed probability of transmitter release, which was common to both glutamate and GABA transmission. Thus, while other recent evidence links alteration of the releasable pool size with changes in p(r), our results suggest the two can be independently manipulated. Selective depression of glutamate release may provide an adaptive mechanism by which neurons limit excitotoxicity. PMID- 15134640 TI - Altered representation of the spatial code for odors after olfactory classical conditioning; memory trace formation by synaptic recruitment. AB - In the olfactory bulb of vertebrates or the homologous antennal lobe of insects, odor quality is represented by stereotyped patterns of neuronal activity that are reproducible within and between individuals. Using optical imaging to monitor synaptic activity in the Drosophila antennal lobe, we show here that classical conditioning rapidly alters the neural code representing the learned odor by recruiting new synapses into that code. Pairing of an odor-conditioned stimulus with an electric shock-unconditioned stimulus causes new projection neuron synapses to respond to the odor along with those normally activated prior to conditioning. Different odors recruit different groups of projection neurons into the spatial code. The change in odor representation after conditioning appears to be intrinsic to projection neurons. The rapid recruitment by conditioning of new synapses into the representation of sensory information may be a general mechanism underlying many forms of short-term memory. PMID- 15134641 TI - Neural correlates of vibrissa resonance; band-pass and somatotopic representation of high-frequency stimuli. AB - The array of vibrissae on a rat's face is the first stage of a high-resolution tactile sensing system. Recently, it was discovered that vibrissae (whiskers) resonate when stimulated at specific frequencies, generating several-fold increases in motion amplitude. We investigated the neural correlates of vibrissa resonance in trigeminal ganglion and primary somatosensory cortex (SI) neurons (regular and fast spiking units) by presenting low-amplitude, high-frequency vibrissa stimulation. We found that somatosensory neurons showed band-pass tuning and enhanced sensitivity to small amplitude stimuli, reflecting the resonance amplification of vibrissa motion. Further, a putative somatotopic map of frequency selectivity was observed in SI, with isofrequency columns extending along the representations of arcs of vibrissae, in agreement with the gradient in vibrissa resonance across the vibrissa pad. These findings suggest several parallels between frequency processing in the vibrissa system and the auditory system and have important implications for detection and discrimination of tactile information. PMID- 15134642 TI - Differential but complementary mnemonic functions of the hippocampus and subiculum. AB - In this study we describe how the hippocampus and subiculum act in concert to encode information in a spatial delayed-nonmatch-to-sample (DNMS) task. This encoding was functionally partitioned between neurons within subiculum and hippocampus to uniquely identify trial-specific information accounting for both spatial and temporal constraints on performance within and between trials. Encoding by subicular neurons in the task was normally accurate and specific, but only if delays were shorter than 15 s, whereas trial-specific information encoded by hippocampal neurons was subject to strong biases from prior trial sequences and was accessible only when delays exceeded 15 s. The two structures operated in a complementary manner to encode information correctly on 75% of all trials using the above strategies. The remaining 25% of trials were at risk due to inherent idiosyncrasies by which hippocampal and subicular neurons encoded information and became errors when the random sequence of trials conflicted with these constraints. PMID- 15134644 TI - Patterns of ongoing activity and the functional architecture of the primary visual cortex. AB - Ongoing spontaneous activity in the cerebral cortex exhibits complex spatiotemporal patterns in the absence of sensory stimuli. To elucidate the nature of this ongoing activity, we present a theoretical treatment of two contrasting scenarios of cortical dynamics: (1) fluctuations about a single background state and (2) wandering among multiple "attractor" states, which encode a single or several stimulus features. Studying simplified network rate models of the primary visual cortex (V1), we show that the single state scenario is characterized by fast and high-dimensional Gaussian-like fluctuations, whereas in the multiple state scenario the fluctuations are slow, low dimensional, and highly non-Gaussian. Studying a more realistic model that incorporates correlations in the feed-forward input, spatially restricted cortical interactions, and an experimentally derived layout of pinwheels, we show that recent optical-imaging data of ongoing activity in V1 are consistent with the presence of either a single background state or multiple attractor states encoding many features. PMID- 15134643 TI - Representation of well-learned information in the monkey hippocampus. AB - In the neocortex, extensive training results in enhanced neuronal selectivity for learned stimuli relative to novel stimuli. This enhanced selectivity has been taken as evidence for learning-related plasticity. Much less is known, in contrast, about the representation of well-learned information in the hippocampus. In this study, we examined the responses of individual hippocampal neurons to well-learned and novel stimuli presented in the context of an associative learning task. There was no difference in the response magnitude or visual response latency of hippocampal neurons to the well-learned and novel stimuli. In contrast, hippocampal neurons responded significantly more selectively to the well-learned stimuli relative to the novel stimuli. These findings show that hippocampal cells, like neocortical cells, show greater selectivity to well-learned stimuli compared to novel stimuli. PMID- 15134645 TI - A deficit in covert attention after parietal cortex inactivation in the monkey. AB - Although the parietal cortex has been repeatedly implicated in controlling attention, the nature and importance of this contribution remain unclear. Here we show that inactivating the lateral intraparietal area in monkeys delays the detection of a visual target located in the contralateral visual field. This effect was observed using different visual scene configurations, e.g., with distractors that differ in number or that differ from the target by a conjunction of shape and color or by a single feature. Since eye movements were not allowed during the searching tasks, these results argue for an unambiguous role of the parietal cortex in the top-down control of attentional deployment in space. PMID- 15134646 TI - Human striatal responses to monetary reward depend on saliency. AB - While the striatum has been implicated in reward processing, an alternative view contends that the striatum processes salient events in general. Using fMRI, we investigated human striatal responses to monetary reward while modulating the saliency surrounding its receipt. Money was maximally salient when its receipt depended on a correct response (active) and minimally salient when its receipt was completely independent of the task (passive). The saliency manipulation was confirmed by skin conductance responses and subjective ratings of the stimuli. Significant caudate and nucleus accumbens activations occurred following the active compared to passive money. Such activations were attributed to saliency rather than the motor requirement associated with the active money because striatal activations were not observed when the money was replaced by inconsequential, nonrewarding stimuli. The present study provides evidence that the striatum's role in reward processing is dependent on the saliency associated with reward, rather than value or hedonic feelings. PMID- 15134647 TI - Conformational constraints for amyloid fibrillation: the importance of being unfolded. AB - Recent reports give strong support to the idea that amyloid fibril formation and the subsequent development of protein deposition diseases originate from conformational changes in corresponding amyloidogenic proteins. In this review, recent findings are surveyed to illustrate that protein fibrillogenesis requires a partially folded conformation. This amyloidogenic conformation is relatively unfolded, and shares many structural properties with the pre-molten globule state, a partially folded intermediate frequently observed in the early stages of protein folding and under some equilibrium conditions. The inherent flexibility of such an intermediate is essential in allowing the conformational rearrangements necessary to form the core cross-beta structure of the amyloid fibril. PMID- 15134648 TI - Functional plasticity and catalytic efficiency in plant and bacterial ferredoxin NADP(H) reductases. AB - Ferredoxin (flavodoxin)-NADP(H) reductases (FNRs) are ubiquitous flavoenzymes that deliver NADPH or low potential one-electron donors (ferredoxin, flavodoxin, adrenodoxin) to redox-based metabolisms in plastids, mitochondria and bacteria. Two great families of FAD-containing proteins displaying FNR activity have evolved from different and independent origins. The enzymes present in mitochondria and some bacterial genera are members of the structural superfamily of disulfide oxidoreductases whose prototype is glutathione reductase. A second group, comprising the FNRs from plastids and most eubacteria, constitutes a unique family, the plant-type FNRs, totally unrelated in sequence with the former. The two-domain structure of the plant family of FNR also provides the basic scaffold for an extended superfamily of electron transfer flavoproteins. In this article we compare FNR flavoenzymes from very different origins and describe how the natural history of these reductases shaped structure, flavin conformation and catalytic activity to face the very different metabolic demands they have to deal with in their hosts. We show that plant-type FNRs can be classified into a plastidic class, characterised by extended FAD conformation and high catalytic efficiency, and a bacterial class displaying a folded FAD molecule and low turnover rates. Sequence alignments supported this classification, providing a criterion to predict the structural and biochemical properties of newly identified members of the family. PMID- 15134649 TI - Biochemical characterization of an inhibitor of Escherichia coli UDP-N acetylmuramyl-l-alanine ligase. AB - UDP-N-acetylmuramyl-l-alanine ligase (MurC) is an essential bacterial enzyme involved in peptidoglycan biosynthesis and a target for the discovery of novel antibacterial agents. As a result of a high-throughput screen (HTS) against a chemical library for inhibitors of MurC, a series of benzofuran acyl-sulfonamides was identified as potential leads. One of these compounds, Compound A, inhibited Escherichia coli MurC with an IC(50) of 2.3 microM. Compound A exhibited time dependent, partially reversible inhibition of E. coli MurC. Kinetic studies revealed a mode of inhibition consistent with the compound acting competitively with the MurC substrates ATP and UDP-N-acetyl-muramic acid (UNAM) with a K(i) of 4.5 microM against ATP and 6.3 microM against UNAM. Fluorescence binding experiments yielded a K(d) of 3.1 microM for the compound binding to MurC. Compound A also exhibited high-affinity binding to bovine serum albumin (BSA) as evidenced by a severe reduction in MurC inhibition upon addition of BSA. This finding is consistent with the high lipophilicity of the compound. Advancement of this compound series for further drug development will require reduction of albumin binding. PMID- 15134650 TI - Mass spectrometry and structural characterization of 2S albumin isoforms from Brazil nuts (Bertholletia excelsa). AB - Proteomic approaches have been used to characterise the main 2S albumin isoforms from Brazil nuts (Bertholletia excelsa). Whilst most isoforms ( approximately 10 discrete protein species) exhibited molecular masses of around 12 kDa with a high amino acid sequence homology, important charge heterogeneity was found, with pIs varying between 4.6 and 6.6, with one >or=7.0. Proteomic analysis showed that these corresponded to a total of six National Center for Biotechnology Information (NCBI) accessions and that three isoforms had been purified to homogeneity corresponding to gi/384327, 112754 and 99609. The latter sequence corresponds to an isoform, previously only identified at the nucleotide sequence level, had a slightly higher molecular weight (13.4 kDa), and with noticeable differences in the primary structure. Proteins corresponding to six different NCBI accessions were identified, the heterogeneity of which had been increased by posttranslational processing. Evidence was found of cyclization of the N-terminal glutamine residue in two isoforms, together with ragged C-termini, indicative of carboxypeptidase activity within the vacuole following posttranslational processing. No evidence of glycosylation was found. Circular dichroism (CD) and Fourier transform-infrared (FT-IR) spectroscopy indicated all the studied isoforms were predominantly alpha-helical in nature, but that the Mr 13400 species was structurally distinct, with a higher proportion of alpha-helical structure. PMID- 15134651 TI - Stepwise construction of triple-helical heparin binding sites using peptide models. AB - The specific localization of the asymmetric form of acetylcholinesterase (AChE) in neuromuscular junctions results from the interaction of its collagen-like tail with heparan sulfate proteoglycans in the synaptic basal lamina. This interaction involves two heparin binding consensus sequences of the form XBBXB, where B is a basic residue, located in the triple-helical collagen tail: GRKGR for the N terminal site and GKRGK for the C-terminal site. To explore the basis of the higher heparin affinity seen for the C-terminal site vs. the N-terminal site, two homologous series of (Gly-Xaa-Yaa)(8) peptides were constructed to model these triple-helical binding sites. Individual tripeptide units from each heparin binding site were introduced in a stepwise fashion into a Gly-Pro-Hyp framework, until the consensus sequence and its surrounding triplets were recreated. As each additional triplet from the binding site is inserted to replace a host Gly-Pro Hyp triplet, the triple-helix stability decreases, and the drop in thermal stability is close to that expected if each Gly-X-Y triplet contributed independently to global stability. CD spectroscopy and calorimetry show the stability of these AChE model peptides is increased by addition of heparin, confirming binding to heparin, and the lack of significant enthalpy change indicates the binding is largely electrostatic in nature. Displacement assays measure the strength of the peptide-heparin interaction, and indicate an inverse correlation between the peptide ability to bind heparin and its thermal stability. The model peptides for the C-terminal binding site show a greater heparin affinity than the peptide models for the N-terminal binding site only when residues surrounding the consensus sequence are included. PMID- 15134652 TI - Complex cooperativity of ATP hydrolysis in the F(1)-ATPase molecular motor. AB - F(1)-ATPase catalyses ATP hydrolysis and converts the cellular chemical energy into mechanical rotation. The hydrolysis reaction in F(1)-ATPase does not follow the widely believed Michaelis-Menten mechanism. Instead, the hydrolysis mechanism behaves in an ATP-dependent manner. We develop a model for enzyme kinetics and hydrolysis cooperativity of F(1)-ATPase which involves the binding-state changes to the coupling catalytic reactions. The quantitative analysis and modeling suggest the existence of complex cooperative hydrolysis between three different catalysis sites of F(1)-ATPase. This complexity may be taken into account to resolve the arguments on the binding change mechanism in F(1)-ATPase. PMID- 15134653 TI - The expression and processing of two recombinant 2S albumins from soybean (Glycine max) in the yeast Pichia pastoris. AB - Soybean seeds contain two 2S albumin storage proteins (AL1 and AL3) which may contribute to their industrial processing quality and allergenicity. We show that these proteins (AL1 and AL3) are well expressed by the methylotrophic yeast Pichia pastoris and that one of the secreted proteins (AL3) has a similar conformation and stability to that purified from soybean seeds. Further, we show that the subunits are post-translationally processed within the same loop region as the native protein but with some differences in the precise sites. This internal processing provides useful information on the endoproteolytic activity in P. pastoris. We also show that, similar to many plant allergens, the 2S albumins from soybean are stable to heat and chemical treatments. PMID- 15134654 TI - Fluorescence analysis of hormone binding activities of wheat germ agglutinin. AB - Wheat germ agglutinin (WGA) from embryos of the monocotyledonous plant Triticum vulgaris (Graminaceae) is a carbohydrate binding protein characterized by high specificity to N-acetyl-d-glucosamine and N-acetyl-d-neuraminic acid. In this study we show that parallel to its carbohydrate binding activities, WGA binds with several orders of magnitude higher affinity adenine, adenine-related cytokinins: kinetin, zeatin and isopentenyl-adenine as well as abscisic and gibberellic acids (K(d) 0.43-0.65 microM). Its interactions with these ligands cause conformational rearrangements in the protein molecules and significant enhancement of the protein tryptophan fluorescence (up to 60%) allowing characterization of the protein-hormone complexes. Dimeric WGA molecules possess two different classes of binding sites for the fluorescent hydrophobic probe 2-(p toluidinyl) naphthalene sulfonic acid (TNS) as suggested by the sigmoid shape of the fluorescence titration curve and the value of the Hill coefficient (n(H) 1.6+/-0.3). The plant hormones displace part of the bound TNS probe and share the higher affinity TNS binding sites. These results characterize WGA as a hormone binding protein. PMID- 15134655 TI - Structural analysis of Salmonella enterica effector protein SopD. AB - Salmonella outer protein D (SopD) is a type III secreted virulence effector protein from Salmonella enterica. Full-length SopD and SopD lacking 16 amino acids at the N-terminus (SopDDeltaN) have been expressed as fusions with GST in Escherichia coli, purified with a typical yield of 20-30 mg per litre of cell culture and crystallized. Biophysical characterization has been carried out mainly on SopDDeltaN. Analytical size exclusion chromatography shows that SopDDeltaN is monomeric and probably globular in aqueous solution. The secondary structure composition, calculated from the CD spectrum, is mixed (38% alpha-helix and 26% beta-strand). Sequence analysis indicates that SopD contains a coiled coil motif, as found in numerous other type III secretion system-associated proteins. This suggests that SopD has the potential for one or more heterotypic protein-protein interactions. Limited trypsin digestion of SopDDeltaN, monitored by both one-dimensional proton NMR spectroscopy and SDS-PAGE, shows that the protein has a large, protease-resistant core domain of 286 amino acid residues. This single-domain architecture suggests that SopD lacks a cognate chaperone. In crystallization trials, SopDDeltaN produced better crystals than either full length SopD or trypsin-digested SopDDeltaN. Diffraction to 3.0 A resolution has so far been obtained from crystals of SopDDeltaN. PMID- 15134656 TI - Implications of the ligandin binding site on the binding of non-substrate ligands to Schistosoma japonicum-glutathione transferase. AB - The binding interactions between dimeric glutathione transferase from Schistosoma japonicum (Sj26GST) and bromosulfophthalein (BS) or 8-anilino-1-naphthalene sulfonate (ANS) were characterised by fluorescence spectroscopy and isothermal titration calorimetry (ITC). Both ligands inhibit the enzymatic activity of Sj26GST in a non-competitive form. A stoichiometry of 1 molecule of ligand per mole of dimeric enzyme was obtained for the binding of these ligands. The affinity of BS is higher (K(d)=3.2 microM) than that for ANS (K(d)=195 microM). The thermodynamic parameters obtained by calorimetric titrations are pH independent in the range of 5.5 to 7.5. The interaction process is enthalpically driven at all the studied temperatures. This enthalpic contribution is larger for the ANS anion than for BS. The strongly favourable enthalpic contribution for the binding of ANS to Sj26GST is compensated by a negative entropy change, due to enthalpy-entropy compensation. DeltaG degrees remains almost invariant over the temperature range studied. The free energy change for the binding of BS to Sj26GST is also favoured by entropic contributions at temperatures below 32 degrees C, thus indicating a strong hydrophobic interaction. Heat capacity change obtained for BS (DeltaC(p) degrees =(-580.3+/-54.2) cal x K(-1) mol(-1)) is twofold larger (in absolute value) than for ANS (DeltaC(p) degrees =(-294.8+/ 15.8) cal x K(-1) mol(-1)). Taking together the thermodynamic parameters obtained for these inhibitors, it can be argued that the possible hydrophobic interactions in the binding of these inhibitors to L-site must be accompanied by other interactions whose contribution is enthalpic. Therefore, the non-substrate binding site (designed as ligandin) on Sj26GST may not be fully hydrophobic. PMID- 15134657 TI - Structure-activity relationships for the beta-hairpin cationic antimicrobial peptide polyphemusin I. AB - The solution structure of polyphemusin I was determined using (1)H-NMR spectroscopy. Polyphemusin I was found to be an amphipathic, beta-hairpin connected by a type I' beta-turn. The 17 low-energy structures aligned very well over the beta-sheet region while both termini were poorly defined due in part to a hinge-like region centred in the molecule about arginine residues 6 and 16. Conversely, a linear analogue, PM1-S, with all cysteines simultaneously replaced with serine was found to be dynamic in nature, and a lack of medium and long range NOEs indicated that this molecule displayed no favoured conformation. Circular dichroism (CD) spectroscopy confirmed that in solution, 50% trifluoroethanol (TFE) and in the presence of liposomes, PM1-S remained unstructured. The antimicrobial activity of PM1-S was found to be 4- to 16-fold less than that of polyphemusin I and corresponded with a 4-fold reduction in bacterial membrane depolarization. Both peptides were able to associate with lipid bilayers in a similar fashion; however, PM1-S was completely unable to translocate model membranes while polyphemusin I retained this activity. It was concluded that the disulfide-constrained, beta-sheet structure of polyphemusin I is required for maximum antimicrobial activity. Disruption of this structure results in reduced antimicrobial activity and completely abolishes membrane translocation indicating that the linear PM1-S acts through a different antimicrobial mechanism. PMID- 15134658 TI - Crystallization and preliminary crystallographic studies of a bifunctional restriction endonuclease Eco57I. AB - Restriction endonuclease Eco57I from Escherichia coli recognizes asymmetric DNA sequence 5'-CTGAAG and has both restriction (DNA cleavage a short distance away from the recognition site) and modification (methylation) activities residing in a single polypeptide chain. Single crystals of wild-type Eco57I ternary complexes with double-stranded DNA and sinefungin, a stimulator of endonuclease activity, were obtained by the vapor diffusion technique and characterized crystallographically for different variants of the DNA component. The best data for the complex with 25-mer DNA were collected to 4.2-A resolution at 100 K using synchrotron radiation. The crystals are orthorhombic, space group P2(1)2(1)2, with a=164.3, b=293.0, c=71.1 A, and contain two to four copies of the protein in the asymmetric unit. PMID- 15134659 TI - Crystallization of glycosylated human BACE protease domain expressed in Trichoplusia ni. AB - Human beta-amyloid precursor protein cleaving enzyme (beta-secretase, or BACE) belongs to the aspartyl protease family, and is responsible for generating the N terminus of beta-amyloid peptide (Abeta). BACE is a type I transmembrane glycoprotein with pre-, pro- and catalytic domains, a short transmembrane helix and a cytoplasmic region. In this study, a truncated form was engineered to produce the authentic catalytic domain of BACE in Trichoplusia ni (High 5) cells. The glycosylated BACE zymogen (proBACE) was secreted into the conditioned medium for facile purification by metal chelate and gel filtration chromatographies. The mature catalytic domain was obtained by a trans cleavage event under acidic conditions and crystallized in the absence of a bound inhibitor. A complete 3.4 A data set was collected on a single orthorhombic crystal with unit cell parameters a=74 A, b=130 A, c=134A. Successful molecular replacement shows two BACE molecules in the asymmetric unit. PMID- 15134660 TI - Allometric scaling of maximal metabolic rate in mammals: muscle aerobic capacity as determinant factor. AB - Maximal metabolic rate (MMR) of mammals scales differently from basal metabolic rate (BMR). This is first shown by scrutinizing data reported on exercise-induced Vo2 max in 34 eutherian mammalian species covering a body mass range of 7 g-500 kg. Vo2 max was found to scale with the 0.872 (+/-0.029, 95% confidence limits 0.813-0.932) power of body mass which is significantly different from the 3/4 power reported for basal metabolic rate. The aerobic scope is higher in athletic than non-athletic species, and it is also higher in large than in small species. Integrated structure-function studies on a subset of 11 species (body mass 20 g 450 kg) show that the variation of Vo2 max with body size is tightly associated with the aerobic capacity of the locomotor musculature: the scaling exponents for Vo2 max, the total volume of mitochondria, and the volume of capillaries are nearly identical. The higher Vo2 max of athletic species is tightly linked to proportionally larger mitochondrial and capillary volumes in animals of the same size class. As a result Vo2 max is linearly related to both total mitochondrial and capillary erythrocyte volumes. We conclude that the scaling of maximal metabolic rate is explained by features and mechanisms different from those determining basal metabolic rate. PMID- 15134661 TI - Hypercapnia selectively attenuates the somato-sympathetic reflex. AB - The effects of hyperoxic hypercapnia (5, 10 or 15% CO2 in O2) on splanchnic sympathetic nerve activity (sSNA) and sympathetic reflexes such as the somato sympathetic reflex or baroreflex were studied in urethane anaesthetised, paralysed, artificially ventilated and vagotomized Sprague-Dawley rats. Hypercapnia caused a small increase in mean arterial blood pressure (MAP) in the 10% CO2 group and a fall in heart rate (HR) in all three groups. sSNA increased in all three groups. Phrenic frequency and amplitude increased during hypercapnia, with frequency adapting back towards baseline during the CO2 exposure. The somato-sympathetic reflex was attenuated in the 5% CO2 group and abolished in the 10 and 15% CO2 groups, whereas there was little effect on the sSNA baroreflex. Hypercapnia significantly affects phrenic nerve activity (PNA), sSNA and selectively inhibits the somato-sympathetic reflex with little effect on the sSNA baroreflex. PMID- 15134662 TI - Effects of warm and cool thermal conditions on ventilatory responses to hyperoxic test in neonates. AB - Body temperature interacts with respiratory control, but it is unclear what sites or mechanisms mediate those interactions. We hypothesized that warm and cool thermal conditions affect the decrease in ventilation (VE) seen during the hyperoxic test (HT), a breathing response believed to reflect the strength of the peripheral chemoreceptor drive. A breath-by-breath analysis during a 30 s HT was performed in eight premature neonates (postconceptional age: 36 +/- 1 weeks) under neutral, warm, and cool thermal conditions. Quiet sleep (QS) and active sleep (AS) were scored by neurophysiological criteria. The VE fall was higher in AS than in QS, and warm and cool conditions significantly enhanced the response only in AS (-24.2 +/- 6.0, -39.1 +/- 9.1, and -37.5 +/- 14.1% in neutral, warm, and cool conditions, respectively). Central control mechanisms of the respiratory chemoreflex may explain the increase in peripheral chemoreceptor drive during AS in response to thermal challenges, which may produce increased breathing instability leading to apnea in early life. PMID- 15134663 TI - Tongue mechanical characteristics and genioglossus muscle EMG in obstructive sleep apnoea patients. AB - The increased genioglossus muscle (GGm) activity seen in obstructive sleep apnoea syndrome (OSAS) may lead to increased fatigability or longer recovery time of the tongue. Maximal force, endurance, and recovery times of the tongue, electromyogram (EMG) absolute value, and EMG spectral analysis of the GGm obtained during submaximal contractions were compared in eight individuals without chronic snoring and eight OSAS patients. Endurance time values were not significantly different between the two groups (P = 0.40). Time to recovery of initial maximal force was significantly greater in the OSAS group (P = 0.01). Final EMG median frequency was significantly higher (P = 0.01) and the final low frequency EMG component smaller in the OSAS patients (P = 0.02). Patients did not have changes in endurance time or fatigability but had longer recovery times and changes in spectral analysis variations. This functional investigation may be helpful in determining the presence of OSAS and the potential contribution of the tongue to pharyngeal obstruction. PMID- 15134664 TI - Cerebral oxygenation during intermittent supramaximal exercise. AB - This study examined cerebral deoxygenation during intermittent supramaximal exercise in six healthy male subjects (age: 27.2 +/- 0.6 years (mean +/- S.E.). The subjects performed seven times exercise at an intensity corresponding to 150% of maximal oxygen uptake (VO2max) on cycle ergometer (30 s exercise/15 s rest). Cerebral oxygenation was measured by near-infrared spectroscopy (NIRS). The peak blood lactate concentration after exercise was 15.3 +/- 0.2 mmol/l. Cerebral oxygenation increased in first repetition compared with at rest (+ 5.7 +/- 0.6 microM; P < 0.05), but then decreased with time. Thus, in the last repetition cerebral oxygenation was - 8.5 +/- 0.4 microM (P < 0.05). There was no significant change in arterial oxygen saturation (99.6 +/- at rest, 98.4 +/- 0.2 at the final set of intermittent exercise), and there was no correlated change in end-tidal CO2 concentration with cerebral oxygenation (P > 0.05). These findings suggest that the fatigue resulting from dynamic severe exercise related to a decrease in the cerebral oxygenation level. PMID- 15134665 TI - Buffering limits plasma HCO3- dehydration when red blood cell anion exchange is inhibited. AB - Theory suggests that HCO3- dehydration in the plasma of rainbow trout is limited by both the absence of carbonic anhydrase (CA) activity and the low non bicarbonate buffer capacity of the plasma (betaplasma). The potential for betaplasma to limit plasma HCO3- dehydration was assessed in rainbow trout in which HCO3- dehydration via the red blood cell (RBC) was inhibited using the anion exchange blocker 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS). DIDS administration reduced the rate of RBC HCO3- dehydration by 68-80% for at least 6h, resulting in the elevation of arterial CO2 tension (PaCO2) by 3.07 +/- 0.45 Torr (N = 6). Addition of bovine CA to the circulation of DIDS-treated trout caused PaCO2 to decrease significantly. This effect was increased significantly in rainbow trout in which betaplasma was elevated experimentally by intravascular injection of N-[2-hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid] (HEPES), supporting the hypothesis that CA-catalysed HCO3- dehydration in the plasma of rainbow trout is limited by proton availability. PMID- 15134666 TI - Contribution of nitric oxide to adaptation of tibetan sheep to high altitude. AB - We examined the effects of endogenous nitric oxide synthase (NOS) inhibition on pulmonary hemodynamics in awake sheep living at low and high altitudes to evaluate the role of NO in adaptation to an hypoxic environment. Unanaesthetized male sheep in three places--Matsumoto, Japan (680 m above sea level), Xing, China (2300 m) and Maxin, China (3750 m)--were prepared for measurements of pulmonary artery (Ppa) and pulmonary vascular resistance (PVR) before and after the NOS inhibition. The non-selective NOS inhibitor, Nw-nitro-l-argine (NLA, 20 mg/kg) was used. Baseline Ppa became elevated with an increase in altitude. After NLA administration, PVR significantly increased in animals of all groups. However, the increase in PVR after NLA in tibetan sheep at 3750 m was significantly higher than those in other groups. We conclude that augmented endogenous NO production may contribute to regulating the pulmonary vascular tone in tibetan sheep (3750 m) adapted to high altitude. PMID- 15134667 TI - Seasonal changes in blood oxygen transport and acid-base status in the tegu lizard, Tupinambis merianae. AB - Oxygen-binding properties, blood gases, and acid-base parameters were studied in tegu lizards, Tupinambis merianae, at different seasons and temperatures. Independent of temperature and pH, blood oxygen affinity was higher in dormant lizards than in those active during the summer. Haematocrit (Hct) and hemoglobin content ([Hb]) were greater in active lizards resulting in a higher oxygen carrying capacity. Nucleoside triphosphate content ([NTP]) was reduced during dormancy, but the ratio between [NTP] and [Hb] remained unchanged. Dormancy was accompanied by an increase in plasma bicarbonate ([HCO-(3)]pl) and an elevation of arterial CO2 partial pressure (PaCO2) and CO2 content in the plasma (CplCO2). These changes in acid-base parameters persist over a broad range of body temperatures. In vivo, arterial O2 partial pressure (PaO2) and O2 content (CaO2) were not affected by season and tended to increase with temperature. Arterial pH (pHa) of dormant animals is reduced compared to active lizards at body temperatures below 15 degrees C, while no significant difference was noticed at higher temperatures. PMID- 15134668 TI - Tomorrow's ethics for today's practice. PMID- 15134669 TI - A human rights framework for midwifery care. AB - This article presents a rights-based model for midwifery care of women and childbearing families. Salient features include discussion of the influence of values on how women are viewed within cultures and societies, universal ethical principles applicable to health care services, and human rights based on the view of women as persons rather than as objects or chattel. Examples of the health impact on women of persistent violation of basic human rights are used to support the need for using a human rights framework for midwifery care--a model supported by codes of ethics, the midwifery philosophy of care, and standards of practice. PMID- 15134670 TI - The interface between legal and ethical issues in reproductive health. AB - In a time of rapid change in the health care delivery system, health care professionals are challenged to fully understand their legal and ethical obligations for practice. The interface between law and ethics as it affects professional practice is complex and confusing. Three areas of legal and ethical interface and their associated history are explored: laws affecting scope of practice and the ethical obligation to provide access to care; laws affecting informed consent and the ethical obligation to respect autonomy; and laws limiting disclosure of personal identifiable information and the ethical obligation to protect privacy. A trend toward the use of statutes to protect and enforce patient rights has emerged, which has altered and limited professional practice. PMID- 15134671 TI - Bridging the gap: teaching ethics in midwifery practice. AB - Teaching is an integral part of all midwives' practice, wherever they work. Midwives face ethical issues of greater or lesser significance throughout their careers. In a multitude of different situations and cultures worldwide midwives confront, on a daily basis, the need to make or help others make critical decisions that will impact lives. Midwives need tools to address this reality constructively, consistently, and respectfully if they are to remain true to themselves. They need also to be able to transmit such skills to the students they teach and the women to whom they offer care. Translating ethical thought and reflection into action--bridging the theory-practice gap--is the perennial challenge. This article presents some of the tools available to professionals who take up the challenge of using ethical thinking to shape best practice in any setting. PMID- 15134672 TI - Ethical considerations related to the inclusion of women in clinical trials. AB - Clinical trials are the primary way the most promising new preventive, diagnostic, therapeutic, and palliative measures move from the basic science laboratory to the bedside. Attracting participants to clinical trials occurs at a painstakingly slow pace, delaying the public's access to new care modalities. Additional ways are needed to increase the public's awareness and understanding of the important role of clinical trials. As key members of the health and social welfare promotion team, nurse-midwives/midwives are well positioned to help advance the public's access to clinical trials information. Generic ethical issues related to human subjects review processes, their specific application to clinical trials, and the impact of recent HIPAA legislation are discussed. PMID- 15134673 TI - Issues in patient education. AB - Nurses and nurse midwives have historically considered patient education one of their most important responsibilities. Increasingly, however, appropriate and comprehensive patient education has become more difficult to accomplish. There are many reasons for this, including the huge influx of clients of varying cultures into virtually all health care systems across the United States, the lack of time available for patient education in tightly scheduled managed care visits, the dearth of educational materials written at appropriate readability levels and/or in languages other than English, and the lack of reimbursement for time spent on patient education. In addition, many providers might not have specific training in the provision of patient education or in the development of appropriate health educational materials for the population they serve. This article discusses these issues, suggesting also that there are ethical dilemmas inherent in the provision of some aspects of patient education. PMID- 15134674 TI - Genetic testing for breast and ovarian cancer susceptibility: a family experience. AB - The purpose of this article is to provide an overview of current knowledge concerning genetic testing for breast and ovarian cancer susceptibility. This overview includes 1) a brief history of genetic testing for breast and ovarian cancer susceptibility, 2) a review of factors that midwives and other health care providers need to consider before offering this type of testing to their clients, 3) management options for clients at high risk for hereditary breast and ovarian cancers, and 4) a discussion of preliminary findings from an ongoing study concerning the family experience of genetic testing, which illustrates some of the ethical issues that emerge for individuals and families during the family experience of genetic testing for breast and ovarian cancer susceptibility. PMID- 15134675 TI - Communicating risk in prenatal genetic testing. AB - Prenatal testing for Down syndrome and neural tube defects has become routine, and testing for other genetic conditions is becoming commonplace. Counseling about these tests involves a discussion of risk information, so pregnant women and their partners can use the information effectively when they make choices about testing. Discussing risk can be challenging, as many individuals, particularly those of lower literacy, have a poor understanding of the numerical concept of risk. Furthermore, whether risk is comprehended accurately or not, it is interpreted by patients in light of their existing knowledge and past experiences. Strategies available to optimize understanding of risk include communication of risk figures as frequencies rather than as probabilities or percentages and explicit discussion of a woman's preconceptions about her risk and about the condition being tested for. PMID- 15134676 TI - Women's experiences of unexpected ultrasound findings. AB - Ultrasound imaging is an important screening and diagnostic tool in prenatal care, but for many couples in Canada and the United States, it is primarily a meaningful social ritual of seeing and meeting their baby. This article examines how women perceive ultrasound when they receive unexpected abnormal ultrasound findings. Drawing from qualitative analysis of semistructured interviews with 42 Canadian women, the article discusses women's reactions to receiving unexpected findings, and their perspectives on disclosure of results and on seeing the impaired fetus. Implications are discussed for practitioners regarding prescan counseling, informed choice, and disclosure of abnormal ultrasound findings. PMID- 15134677 TI - Options counseling: techniques for caring for women with unintended pregnancies. AB - An unplanned pregnancy is a crisis in a woman's life. She may request assistance from the health care provider in reviewing her options of becoming a parent, continuing the pregnancy and placing her baby for adoption, or terminating the pregnancy. To facilitate the decision-making process for women facing these choices, the clinician must first examine her own values and biases. To aid the clinician in providing nonjudgmental and nondirective counseling, this article provides factual information about abortion and adoption, values clarification exercises, and concrete approaches to help women examine their beliefs. PMID- 15134678 TI - Examining an ethical dilemma: a case study in clinical practice. AB - When clients and health care providers differ in their understanding of what is right or wrong, an ethical dilemma may arise. Such dilemmas occur in everyday clinical practice. Health care providers have the professional responsibility to analyze these dilemmas. A clinical case study of an ethical dilemma that occurred in a cross-cultural context is examined. The language of the client and provider differed, and no interpreter service was available. Given these conditions, the provider's ethical dilemma was whether, and if so how, to give safe, satisfying care that respected the needs of a client with limited English proficiency. Measuring the morality of the provider's decisions and actions using Rawls' ethical theory of social justice finds deficits. A 10-step Bioethical Decision Making Model by Thompson is used to demonstrate one method for analyzing the moral dimension of a clinical scenario focusing on the decisions and actions taken by a midwife. Scrutinizing ethically challenging clinical encounters will result in better understanding of the moral dimensions of practice. PMID- 15134679 TI - Context and process of informed consent for pharmacologic strategies in labor pain care. AB - The care of women experiencing labor pain often challenges midwives to provide ethical informed consent for the pharmacologic strategies common in most of America's childbearing environments. A systematic approach to this clinical dilemma begins with a conceptual understanding of the origins of labor pain stimuli, factors affecting their central nervous system processing, and the differentiation of the concepts of pain, suffering, and comfort. These concepts can be integrated into a labor pain care, rather than pain management, model for clinical practice. Because most midwives provide care for laboring women in acute care hospitals, the midwife must also understand how the standards of the Joint Commission on Accreditation of Healthcare Organizations influence the behavior of other professionals in the health care environment for laboring women and the process of informed consent. A systematic approach to informed consent for pharmacologic strategies for labor pain care strategies should begin during pregnancy and includes full, unbiased disclosure of the nature, benefits, risks, side effects, and efficacy of all methods that are available in the chosen birth setting. PMID- 15134680 TI - The dangers of geneticism. PMID- 15134681 TI - A code of ethics for midwives. PMID- 15134682 TI - Prophylactic use of acyclovir reduces recurrence in pregnant women at term. PMID- 15134683 TI - Emergency contraception is not associated with reduction in contraceptive use or an increase in adverse health outcomes. PMID- 15134684 TI - Asthma during pregnancy not associated with adverse perinatal outcomes. PMID- 15134685 TI - Contraceptive options for women with factor V Leiden. PMID- 15134686 TI - Role of malpractice crisis in midwifery job market. PMID- 15134689 TI - TMS for ALS: why and why not. PMID- 15134690 TI - Uncovering the mechanism(s) of action of deep brain stimulation: activation, inhibition, or both. AB - High-frequency deep brain stimulation (DBS) of the thalamus or basal ganglia represents an effective clinical technique for the treatment of several medically refractory movement disorders. However, understanding of the mechanisms responsible for the therapeutic action of DBS remains elusive. The goal of this review is to address our present knowledge of the effects of high-frequency stimulation within the central nervous system and comment on the functional implications of this knowledge for uncovering the mechanism(s) of DBS. Four general hypotheses have been developed to explain the mechanism(s) of DBS: depolarization blockade, synaptic inhibition, synaptic depression, and stimulation-induced modulation of pathological network activity. Using the results from functional imaging, neurochemistry, neural recording, and neural modeling experiments we address the general hypotheses and attempt to reconcile what have been considered conflicting results from these different research modalities. Our analysis suggests stimulation-induced modulation of pathological network activity represents the most likely mechanism of DBS; however, several open questions remain to explicitly link the effects of DBS with therapeutic outcomes. PMID- 15134691 TI - One set of sounds, two tonotopic maps: exploring auditory cortex with amplitude modulated tones. AB - OBJECTIVE: The possibility of simultaneously observing activation of primary and secondary auditory cortices has been demonstrated by Engelien et al. [Hear Res 2000;148:153-60]. METHODS: Such a dual monitoring by means of neuromagnetic recordings can be achieved when a subject is stimulated by brief pulses of 40Hz modulated tones. Depending on the frequency filter applied, either the steady state field (SSF) or the N1m can be extracted from the evoked magnetic field complex. RESULTS: Using this "combined" (two-maps) paradigm with 4 carrier frequencies, we show that it is possible to synchronously screen two tonotopic maps--one map each reflected either by the SSF or the N1m. Indicators are the systematic variation in the location (higher frequencies are more posterior) and orientation (higher frequencies oriented differently in the sagittal plane) of the equivalent current dipole (ECD). These parameters were compared with those obtained from "classic" (one map) paradigms in which either a pure tone elicits an N1m or a 40 Hz continuous (3 s) stimulation produces an SSF. Overall the results were similar, however, systematic differences between the paradigms were found for ECD localization, dipole strength, amplitude, and phase. CONCLUSIONS AND SIGNIFICANCE: One possible interpretation of these results is that different tonotopically arranged cortical fields were involved in the generation of the components. PMID- 15134692 TI - Effects of low frequency and low intensity repetitive paired pulse stimulation of the primary motor cortex. AB - OBJECTIVE: Following a previous report [Bestmann et al. Clin Neurophysiol 2004;115:755-64] that pairs of subthreshold pulses of transcranial magnetic stimulation (TMS) can show temporal summation, we explored whether repeated application of pairs of stimulation could produce long-lasting after effects on the excitability of the human motor cortex. METHODS: Twelve healthy subjects received 25 min repetitive paired pulse magnetic stimulation (paired rTMS) given at a frequency of about 0.6 Hz over the left primary motor cortex (500 paired stimuli in total). The interval between the paired stimuli was 3 ms and the intensity of both stimuli was 80% of active motor threshold. The resting and active motor threshold, MEP recruitment curve, short interval intracortical inhibition (SICI) and facilitation, and the duration of the cortical silent period (SP) were tested for the right first interosseous muscle (FDI) before and two times after the end of 25 min paired rTMS. RESULTS: Prolonged subthreshold paired rTMS produced a significant decrease in excitability in the corticospinal projection to FDI: resting motor threshold was significantly increased and MEP recruitment was significantly decreased, SICI was significantly increased at 2 and 4 ms and the SP was significantly increased in duration. CONCLUSIONS: Prolonged low frequency paired rTMS at subthreshold intensity can modulate cortical excitability by producing inhibitory effects that outlast the period of stimulation. PMID- 15134693 TI - Modulation of motor cortex excitability after upper limb immobilization. AB - OBJECTIVE: To examine the mechanisms of disuse-induced plasticity following long term limb immobilization. METHODS: We studied 9 subjects, who underwent left upper limb immobilization for unilateral wrist fractures. All subjects were examined immediately after splint removal. Cortical motor maps, resting motor threshold (RMT), motor evoked potential (MEP) latency and MEP recruitment curves were studied from abductor pollicis brevis (APB) and flexor carpi radialis (FCR) muscles with single pulse transcranial magnetic stimulation (TMS). Paired pulse TMS was used to study intracortical inhibition and facilitation. Compound muscle action potentials (CMAPs) and F waves were obtained after median nerve stimulation. In 4/9 subjects the recording was repeated after 35-41 days. RESULTS: CMAP amplitude and RMT were reduced in APB muscle on the immobilized sides in comparison to the non-immobilized sides and controls after splint removal. CMAP amplitude and RMT were unchanged in FCR muscle. MEP latency and F waves were unchanged. MEP recruitment was significantly greater on the immobilized side at rest, but the asymmetry disappeared during voluntary muscle contraction. Paired pulse TMS showed an imbalance between inhibitory and excitatory networks, with a prevalence of excitation on the immobilized sides. A slight, non-significant change in the strength of corticospinal projections to the non-immobilized sides was found. TMS parameters were not correlated with hand dexterity. These abnormalities were largely normalized at the time of retesting in the four patients who were followed-up. CONCLUSIONS: Hyperexcitability occurs within the representation of single muscles, associated with changes in RMT and with an imbalance between intracortical inhibition and facilitation. These findings may be related to changes in the sensory input from the immobilized upper limb and/or in the discharge properties of the motor units. SIGNIFICANCE: Different mechanisms may contribute to the reversible neuroplastic changes, which occur in response to long-term immobilization of the upper-limbs. PMID- 15134694 TI - Multivariate AR modeling of electromyography for the classification of upper arm movements. AB - OBJECTIVE: We compared the performance of two feature extraction methods for multichannel electromyography (EMG) based arm movement classification. One method was to use a scalar autoregressive model (sAR) for each channel. Another was to model all channels as a whole by a multivariate AR model (mAR). METHODS: The classified arm movements included elbow flexion, elbow extension, forearm pronation and internal shoulder rotation. Six-channel bipolar EMG signals were collected from four electrodes fixed on the biceps, triceps, brachioradialis and deltoid. Fifteen two-channel and four three-channel configurations were formed out of these six-channel signals for a comparison of different channel combinations. Leave-one-out cross-validation was adopted for evaluating the classification performance using a parametric statistical classifier. RESULTS: We processed a total of 216 EMG segments obtained from repeated 18 performances by three normal subjects. mAR model based feature set achieved a better classification accuracy than sAR did for each configuration. Moreover, significance of improvement was greater than 0.95 for those configurations which consisted of EMG channels that were close spatially. CONCLUSIONS: The stronger the cross-correlation among EMG channels the more improvement of classification accuracy one would expect from using a mAR model. PMID- 15134695 TI - Glutamate evoked neck and jaw muscle pain facilitate the human jaw stretch reflex. AB - OBJECTIVE: Although pain and neuromuscular function are clearly linked in several clinical conditions manifested in the craniofacial and cervical regions, it is unclear if pain in these regions influences reflexly evoked activity in the jaw or neck muscles in humans. The aim of the present study was to test the effects of glutamate-evoked jaw or neck muscle pain on the jaw stretch reflex recorded in both jaw and neck muscles. METHODS: Nineteen healthy men participated in the study. Electromyographic (EMG) recordings were made from the left masseter (MAL) and right masseter (MAR) muscles and the right sternocleidomastoid (SCM) and splenius (SP) muscles. Glutamate (1 M) or isotonic saline was injected into the MAR or right SP in random order and then the other solution was injected 1-3 weeks later. Pain intensity was scored on a 10 cm visual analogue scale. Stretch reflexes were evoked by standardized jaw stretches before, during and 15 min after the end of the experimental muscle pain. Twenty trials were averaged in each condition. RESULTS: Pain evoked by MAR or SP glutamate injections was associated with a significant increase in the stretch reflex amplitude recorded in both MAR and SCM. The onset and offset times and duration of the stretch reflex did not change in any muscle during the various pain conditions. Injection of isotonic saline into the MAR or SP did not produce any significant change in the reflex parameters in any of the muscles. CONCLUSIONS: The results indicate the close interplay between the craniofacial and cervical regions in the neuromuscular changes that may result from musculoskeletal pain in either region. SIGNIFICANCE: The changes in neuromuscular activity documented in this study may be involved in the clinical occurrence of altered muscle activity in the orofacial and cervical regions as a result of deep tissue trauma and pain. PMID- 15134696 TI - Effects of loading and unloading of lower limb joints on the soleus H-reflex in standing humans. AB - OBJECTIVE: To investigate the effects of loading and unloading of the lower limb joints on the soleus H-reflex in standing humans. METHODS: H-reflexes were elicited in the soleus muscle in subjects standing on a force platform in a water tank under the following loading conditions of the ankle and knee joints: control condition; reduced loads of -10 and -20 N; imposed loads of 10 and 20 N. The joint loading was altered by changing the combinations of buoys and weights attached to the lower limb segments, while total body weight was kept constant. RESULTS: As the ankle- or knee-joint load was reduced, the H-reflex was significantly enhanced compared to that under the control condition. In contrast, the H-reflex was decreased as the ankle- or knee-joint load was increased. In both cases, similar levels of background activity were recorded. CONCLUSIONS: The present results suggest that joint afferents might mediate the suppression of the soleus H-reflex in standing humans. However, the identification of the receptors and/or the mechanisms cannot be addressed under the current experimental set up. SIGNIFICANCE: The results of this study give some basic insights into reflex control in an upright posture. PMID- 15134697 TI - Handedness is mainly associated with an asymmetry of corticospinal excitability and not of transcallosal inhibition. AB - OBJECTIVE: The study aims to compare transcallosal inhibition (TI), as assessed by the paired-pulse transcranial magnetic stimulation (TMS) technique, in a sample of right-handed subjects (RH) and left-handed subjects (LH). Motor thresholds (MTs) and motor evoked potential (MEP) amplitudes were also measured in the two groups, as an index of corticospinal activity. METHODS: Thirty-two normal subjects (16 RH and 16 LH) were recorded with a paired-pulse TMS paradigm (intensity of both pulses=120% of MT). The inter-stimulus intervals (ISIs) were 2, 4, 6, 8, 10, 12, 14, 16, 18, and 20 ms for both motor cortices, and MEP responses were recorded from the abductor digiti minimi muscles. RESULTS: Both groups showed a clear TI centred around the 12 ms ISI, but no difference was found as a function of handedness or of hemisphere. On the other hand, the two groups differed in terms of corticospinal activity, since the hand motor dominant hemisphere had lower MTs than the non-dominant one in LH, and larger MEP amplitudes for the right hand were found in RH. CONCLUSIONS: Results point to a functional asymmetry of the motor cortex on the hand-dominant versus the non dominant hemisphere, while handedness does not seem associated with functional differences in callosal inhibition, as measured by the inter-hemispheric paired pulse TMS technique. PMID- 15134698 TI - Surface EMG recording of heteronymous reflex excitation of semitendinosus motoneurones by group II afferents. AB - OBJECTIVE: A potent stretch reflex originates in muscle spindle secondary endings and is mediated by group II muscle afferents. Electrical stimulation of peripheral nerve could also induce group II facilitation, as found in single motor unit recordings for various nerve-muscle combinations in man. The aim of the present investigation was to use surface electromyogram (EMG) to record and to quantify heteronymous excitation of semitendinosus (ST) motoneurones by group II afferents of the tibial nerve. METHODS: This study included 20 healthy subjects. The conditioning stimulation of the tibial nerve was delivered at the popliteal fossa at two intensity levels and for two levels of ST contraction. EMG activity was recorded by surface electrodes placed over the ST muscle, and the averaged and rectified ST EMG signal was analyzed. RESULTS: We observed 4 periods of interest: a first period of prestimulus background EMG activity; a second period of early reinforcement of ST EMG activity starting 30 ms poststimulus and lasting for 35-45 ms; a third period of relative inhibition of ST contraction; a fourth period of late reinforcement of poststimulus ST EMG activity. CONCLUSIONS: We hypothesized that the early period of facilitation was mainly related to heteronymous excitation of ST motoneurones by group II afferents on the basis of the following observations: (i) the amount of facilitation increased for a high stimulus intensity level, which was suitable for group II recruitment; (ii) the onset latency of this facilitation was consistent with the theoretically calculated latency of a group II-mediated reflex. SIGNIFICANCE: Surface EMG recording offers various advantages compared to single motor unit recording to study group II facilitation in the ST muscle. This technique could be applied in the future to confirm that heteronymous reflex excitation by group II afferents is enhanced in spastics and plays an important role in the development of lower limb spasticity. PMID- 15134699 TI - Inhibitory processing during the Go/NoGo task: an ERP analysis of children with attention-deficit/hyperactivity disorder. AB - OBJECTIVE: Previous event-related potential (ERP) research on inhibitory functioning in attention-deficit/hyperactivity disorder (AD/HD) has often failed to use tasks which optimally assess inhibition. We report on an improved measure of inhibitory function, involving inhibition of a prepotent response, in children with AD/HD. METHODS: Twelve males with AD/HD and 12 control males, aged 7-12 years, completed a cued Go/NoGo task where Go stimuli were presented on 70% of trials. ERP and behavioural measures were collected, together with reading, spelling and full-scale IQ scores. RESULTS: The behavioural performance of children with AD/HD was not significantly different from normal controls, although children with AD/HD made faster responses and more errors. Group differences were apparent in the early processing components (P1, N1, P2) of responses to Warning, Go and NoGo stimuli. For the frontally maximal N2, a NoGo>Go effect was found, consistent with previous work linking this component with inhibitory processing. In control children this effect was particularly strong in the right frontal region, while children with AD/HD showed a much larger NoGo>Go effect, and an earlier N2 peak, than controls, with a focal shift to the left frontal region. CONCLUSIONS: Compared with normal controls, children with AD/HD demonstrate early stimulus processing atypicalities, suggesting problems with sensory registration and identification of stimuli. Further, N2 results suggest that children with AD/HD must trigger the inhibition process earlier and more strongly than controls to perform at a comparable behavioural level. SIGNIFICANCE: The results support the theory that behavioural inhibition is deficient in AD/HD, as children with AD/HD show abnormalities in inhibitory ERP components relating to the effort involved in inhibiting a prepotent response. PMID- 15134700 TI - EEG synchronization likelihood in mild cognitive impairment and Alzheimer's disease during a working memory task. AB - OBJECTIVE: Synchronization likelihood analysis of resting state EEG has shown that cognitive dysfunction in Alzheimer's disease (AD) and its precursor mild cognitive impairment (MCI) are associated with a loss of functional connectivity in high (upper alpha and beta) frequency bands. Working memory tasks are known to change functional connectivity, but it is unknown whether this increases the differences between AD, MCI and healthy controls. Our objective was to investigate the behavior of synchronization likelihood of multichannel EEG in AD, MCI and cognitively healthy controls, both at rest and during a working memory task. METHODS: EEGs (200 Hz sample frequency, 21 channels, average reference) were recorded at rest as well as during a visual working memory task in 14 patients with AD according to the NINCDS-ADRDA criteria (mean age 76.4; SD 13.6), 11 patients with MCI according to the criteria of Petersen (mean age 78.4; SD 6.4) and 14 with subjective memory complaints but no demonstrable memory disturbance (mean age 61.6; SD 26.6). The synchronization likelihood was computed over 19 channels, comparing each channel with all the other channels for the 0.5 4, 4-8, 8-10, 10-12, 12-30, 30-50 Hz frequency bands. RESULTS: The synchronization likelihood was significantly decreased in the upper alpha (10-12) and beta (12-30) bands in AD compared to persons with subjective memory complaints. The working memory task scores strongly correlated with Mini-Mental State Examination scores. During the working memory task the synchronization likelihood was significantly higher in MCI compared to the control subjects in the lower alpha band (8-10 Hz). CONCLUSIONS: Decrease of beta band synchronization occurs in mild AD, both in a resting condition and during a working memory task. SIGNIFICANCE: Decrease of beta band synchronization in mild AD is a robust finding. The present study confirms our findings in a different cohort of patients, using alternative frequency bands. The diagnostic value of the synchronization likelihood in AD and MCI needs to be further established. PMID- 15134701 TI - Executive hypercontrol in obsessive-compulsive disorder: electrophysiological and neuropsychological indices. AB - OBJECTIVE: Neuropsychological, brain imaging and electrophysiological research have consistently shown a dysfunction of fronto-striato-thalamic pathways in subjects with obsessive-compulsive disorder (OCD). The functional meaning of the observed dysfunction in the pathogenesis of OCD is still debated. In the present study the hypothesis that it might be related to a hyperactive executive control is explored by means of neuropsychological and electrophysiological measures. METHODS: Multilead quantitative EEG (QEEG) characteristics and neuropsychological performance on tests exploring executive functions, attention, short-term memory and the ability to learn supraspan recurring sequences were investigated in 32 drug-free patients with DSM-IV OCD. Multilead QEEG characteristics were also investigated in 32 healthy controls, matched with patients for age, gender and handedness. RESULTS: A decrease of the slow alpha-band power in OCD as compared to healthy subjects was observed. A significant negative correlation between the slow alpha-band power and the time to complete a neuropsychological test exploring executive functions was found: the more reduced the slow alpha-band power, the slower the performance on this test. CONCLUSIONS: The topographic distribution of the observed QEEG abnormalities, as well as their correlations with neuropsychological indices, suggest an increased activation of frontal networks in OC patients. SIGNIFICANCE: Study findings support the presence of a hyperactivity of attention/executive control mechanisms in obsessive-compulsive patients. PMID- 15134702 TI - A common N400 EEG component reflecting contextual integration irrespective of symbolic form. AB - OBJECTIVE: Electroencephalographic (EEG) waves modulated by context have been identified about 400 ms after presentation of a new semantic stimulus, such as a word or a number, within a prior context. However, it is not known if any component of these waves arises from a common brain system activated by different symbolic forms. METHODS: Multichannel EEG recordings were performed in 10 healthy subjects during the presentation of lexical and numerical series with congruent and incongruent endings. EEG was analysed using a combination of independent component and cluster analysis. RESULTS: Contextual integration of semantic stimuli elicited a negative independent component at around 400 ms that shared the same pattern of spatio-temporal covariation across numerical series and sentences within single subjects. This independent component was bigger following incongruent endings. CONCLUSIONS: These data provide evidence that one element of the activity contributing to the N400 is common to different symbolic forms. SIGNIFICANCE: One component of the brain systems evaluating the semantic inter relationship of new stimuli with prior context may be common to different symbolic forms. PMID- 15134703 TI - The old switcheroo: when target environmental sounds elicit a novelty P3. AB - OBJECTIVE: Novel environmental sounds that are task-irrelevant in the novelty oddball paradigm elicit the novelty P3 or P3a with a fronto-central scalp distribution, while pure tone task-relevant stimuli elicit a P3 with a posterior topography (P3b). To determine whether stimulus nature or its function in the task modulates scalp topography, the role of the two types of stimuli during the novelty oddball task was reversed. METHODS: Brain electrical activity was recorded while 12 young adults listened to frequent tones, infrequent tones, and infrequent environmental sounds. Subjects were not informed about the infrequent tones, but were instructed to press a reaction time (RT) button when they heard the infrequent environmental sounds. RESULTS: Despite the "novelty" nature of the tones, they were associated with a posterior scalp distribution typical of the P3b. Similarly, despite the "target" nature of the environmental sounds, they were associated with a fronto-central scalp topography typical of the P3a. CONCLUSIONS: These data suggest that the elicitation of a novelty P3 depends not only on task demands but also on the physical nature of the stimulus. SIGNIFICANCE: The brain's orienting response is modulated by the contextual environment as well as the physical properties of the eliciting stimulus. PMID- 15134704 TI - EEG spectral analysis of wakefulness and REM sleep in high functioning autistic spectrum disorders. AB - OBJECTIVE: The aim of this study was to investigate the involvement of temporo occipital regions in the pathophysiology of autistic spectrum disorders (ASD) by using REM sleep and waking EEG. METHODS: The EEG recordings of 9 persons with ASD and 8 control participants were recorded using a 12-electrode montage. Spectral analysis (0.75-19.75 Hz) was performed on EEG activity recorded upon two activated states: REM sleep and wakefulness. RESULTS: During REM sleep, persons with ASD showed a selective, significantly lower absolute beta (13.0-19.75 Hz) spectral amplitude over the primary (O(1), O(2)) and associative (T(5), T(6)) cortical visual areas compared to controls. Persons with ASD showed significantly higher absolute theta (4.0-7.75 Hz) spectral amplitude over the left frontal pole region (Fp1) compared to controls during evening wakefulness, but not during morning wakefulness. SIGNIFICANCE: The results of waking EEG are consistent with previously reported observations of neuropsychological signs of frontal atypicalities in ASD; results from REM sleep are the first EEG evidence to support the hypothesis of abnormal visuoperceptual functioning in ASD. Altogether, these results point toward atypical thalamo-cortical mechanisms subserving the neural processing of information in ASD. PMID- 15134705 TI - Alcoholism risk, tobacco smoking, and P300 event-related potential. AB - OBJECTIVE: The P300 event-related potential (ERP) is sometimes larger for individuals at low- compared to high-risk for alcoholism. These effects are inconsistent, and how P300 is affected by tobacco smoking in the context of alcoholism risk is unknown. The present study used P300 to examine the inter relationship between alcoholism heritability and smoking status. METHODS: P300 was elicited with a visual discrimination task from young adults at low- and high risk for alcoholism. Half of the subjects in each risk category reported that they did not smoke cigarettes, and the other half reported that they smoked regularly, with equal numbers of male and female subjects assessed. ERPs were recorded, and subjects were instructed to respond only to an infrequently presented target stimulus that occurred in a series of standard and distracter stimuli. RESULTS: P300 amplitude from the target stimuli was larger for the low risk compared to high-risk subjects overall. However, smoking status demonstrated even stronger effects, with non-smokers producing consistently larger component amplitudes than smokers and accounting for more variance than alcoholism risk. These group factors also significantly affected P300 scalp topography. No reliable alcoholism risk or smoking group effects were obtained for the ERPs from the other stimuli. CONCLUSIONS: The findings suggest that P300 measures of alcoholism risk in young adults are moderated by smoking status. Theoretical implications are discussed. PMID- 15134706 TI - Anesthesia affects olfaction and chemosensory event-related potentials. AB - OBJECTIVE: Olfactory and trigeminal systems interact and contribute to the perception of odorants. This study was aimed at investigating the effect of local anesthesia on olfaction. METHODS: One percent of tetracaine on a cotton swab was applied intranasally at three different locations in 20 volunteers and 4% of lidocaine was applied to the olfactory cleft in a head-down position. Before and after anesthesia, self-assessment, psychometric testing and olfactory event related potentials [OERPs, using H(2)S and phenyl ethyl alcohol (PEA)], and chemosomatosensory event-related potentials (CSSERPs, using CO(2)) were examined. RESULTS: Anesthesia at all four locations significantly lowered the perceived self-assessment of olfaction, while using the cotton swab only anesthesia in the middle meatus elevated threshold (P = 0.020), lowered discrimination (P = 0.015), and prolonged OERP (PEA, P = 0.008; H(2)S, P = 0.016), as well as CSSERPs latencies (CO(2), P = 0.020). However, complete temporary anosmia was only achieved after applying 4% lidocaine into the olfactory cleft. CONCLUSIONS: Intranasal anesthesia applied with a swab reduced self-assessment of olfaction but was unable to produce anosmia. Psychometric test results were concordant with changes in chemosensory event-related potentials. SIGNIFICANCE: Temporary anosmia is technically difficult to achieve but could be demonstrated for the first time using local anesthesia. Even though anesthesia influences self-assessment, measurable olfactory function can remain unchanged. PMID- 15134707 TI - Brain-stem components of high-frequency somatosensory evoked potentials are modulated by arousal changes: nasopharyngeal recordings in healthy humans. AB - OBJECTIVE: Until now, the demonstration that early components of high-frequency oscillations (HFOs) evoked by electrical upper limb stimulation are generated in the brain-stem has been based on the results of scalp recordings. To better define the contribution of brain-stem components to HFOs building, we recorded high-frequency somatosensory evoked potentials (SEPs) in 6 healthy volunteers by means of a nasopharyngeal (NP) electrode. Moreover, since HFOs are highly susceptible to arousal fluctuations, we investigated whether eyes opening can influence HFOs at this level. METHODS: We recorded right median nerve SEPs from the ventral surface of the medulla by means of a NP electrode as well as from the scalp, in 6 healthy volunteers under two different arousal states (eyes opened versus eyes closed). SEPs have been further analyzed after digital narrow bandpass filtering (400-800 Hz). RESULTS: NP recordings demonstrated in all subjects a well-defined burst, occurring in the same latency window of the low frequency P13-P14 complex. Eyes opening induced a significant amplitude increase of the NP-recorded HFOs, whereas scalp-recorded HFOs as well as low-frequency SEPs remained unchanged. CONCLUSIONS: Our findings demonstrate that slight arousal variations induce significant changes in brain-stem components of HFOs. According to the hypothesis that HFOs reflect the activation of central mechanisms, which modulate sensory inputs depending on variations of arousal state, our data suggest that this modulation is already effective at brain-stem level. PMID- 15134708 TI - Simultaneous modulation of the exteroceptive suppression periods in the trapezius and temporalis muscles by experimental muscle pain. AB - OBJECTIVE: To investigate whether the exteroceptive suppression (ES) periods in the trapezius and temporalis muscles could be simultaneously modulated by either experimental trapezius muscle pain or temporalis muscle pain. METHODS: Fourteen healthy volunteers participated in two sessions with randomised injections of either hypertonic or isotonic saline into the right trapezius muscle or right temporalis muscle. The pain intensity was continuously scored on a 10 cm electronic visual analogue scale. During muscle contraction, the early (ES1) and late (ES2) reflex in the temporalis and late (ES2) reflex in the trapezius elicited by electrical stimulation of the right infraorbital nerve were recorded pre-, during-, and post-injection. RESULTS: Hypertonic saline injection induced local muscle pain and referred pain with the maximal pain intensity of 6.3+/-0.5 cm in the right trapezius and 6.7+/-0.6 cm in the right temporalis muscles. Injection of either hypertonic or isotonic saline into the right trapezius muscle was associated with significantly shortened ES2 duration and offset time in the injected right trapezius muscle. Only injection of hypertonic saline into the right trapezius muscle could significantly reduce the ES2 offset time and decrease the degree of suppression of ES2 in the right temporalis and left trapezius muscles. Injection of either hypertonic or isotonic saline into the right trapezius muscle evoked no changes in ES2 parameters in the left temporalis muscle. Injection of hypertonic saline, but not isotonic saline, into the right temporalis muscle caused a significantly shorter ES2 offset time and shorter ES2 duration bilaterally in the temporalis and trapezius muscles. CONCLUSIONS: The findings of the present study provide evidence for a bi-directional effects reflecting convergence of muscle afferents from the trigeminal and upper cervical neural systems in humans, which may partly underlie the manifestations of pain referral between these two areas. PMID- 15134709 TI - Interhemispheric functional synchronization at the first step of visual information processing in humans. AB - OBJECTIVE: We examined the interhemispheric functional synchronization of the visual cortex using coherence (Coh) analysis. METHODS: Achromatic or isoluminant chromatic sinusoidal grating stimuli were presented to each hemifield at a rate of 8 reversals/s to record steady-state visual-evoked potentials (S-VEPs) in 10 healthy subjects. Four recording electrodes were placed at O1, O2, P3 and P4, referred to an electrode at Cz. A total of 50 responses of 1 s epoch were averaged, and were subjected to discrete fast Fourier transforms to yield the amplitude and phase of the 8 Hz component. Ordinary and partial Coh values were also calculated. RESULTS: For both achromatic and chromatic stimuli, the 8 Hz amplitudes of O1 and O2 were significantly larger than those of P3 and P4 without any significant difference between O1 and O2. The phase lag between O1 and O2 was approximately 30 degrees (latency shift 10.4 ms). Partial Coh between O1 and O2 at 8 Hz was significantly greater than that of the unstimulated condition, and this was only observed at 8 Hz. CONCLUSIONS: These results suggest that interhemispheric synchronization in the occipital area occurs despite the nature of the visual stimuli. Therefore, the activation of interhemispheric connection is important for the early stage of the visual information processing. SIGNIFICANCE: Our results indicate that the first step of the visual information processing requires interhemispheric functional synchronization. PMID- 15134711 TI - Characteristics of scalp electrical fields associated with deep medial temporal epileptiform discharges. AB - OBJECTIVE: To determine scalp characteristics of epileptiform discharges arising from medial temporal structures (MT). METHODS: Signal-to-noise ratio was increased by averaging simultaneous recordings from intracranial and scalp electrodes synchronised on discharges recorded by foramen ovale (FO) electrodes. The topography, amplitude and distribution of averaged scalp signals were analysed. RESULTS: Four thousand three hundred and twenty-seven discharges from 20 patients were averaged into 77 patterns. Before averaging, only 9% of discharges were detectable on the scalp without the need of simultaneous FO recordings (SED). A further 72.3% of discharges fell into averaged patterns that could be detected on the scalp as small transients before or after averaging (STBA or STAA). In 18.7% of discharges, no scalp signal was seen after averaging. Whereas most SED patterns had largest amplitude on the scalp at anterior temporal electrodes, STBA and STAA patterns showed greater variability and more widespread scalp fields, suggesting a deeper source. Dipole source localisation modelled the majority of SED patterns as radial dipoles located just behind the eye. In contrast, dipoles corresponding to STBA or STAA patterns showed greater variability in location and orientation and tended to be located at MT. CONCLUSIONS: SED patterns seem to arise from widespread subtemporal and/or superficial neocortical activation, generating EEG fields that are distorted by the high electrical conductivity of anterior cranial foramina. In contrast, STBA and STAA patterns represent electrical fields from neuronal activity more restricted to MT, that reach the scalp highly attenuated by volume-conduction and less distorted by cranial foramina. SIGNIFICANCE: Low amplitude scalp signals can be related to MT activity and must be taken into consideration for the diagnosis of temporal lobe epilepsy, pre-surgical assessment and for valid modelling of deep sources from the scalp EEG and magnetoencephalogram. PMID- 15134710 TI - Photosensitive absence epilepsy with myoclonias and heterozygosity for succinic semialdehyde dehydrogenase (SSADH) deficiency. AB - OBJECTIVE: Succinic semialdehyde dehydrogenase (SSADH) deficiency is a neurometabolic disorder characterized by excessive GABA levels and seizures. There has been no clinical phenotype described to date with heterozygosity for SSADH deficiency. METHODS: A patient heterozygous for SSADH deficiency presented with absence and myoclonic seizures. EEG monitoring and enzymatic, metabolic, and molecular studies for SSADH were obtained on the patient and family members. RESULTS: EEG recordings yielded generalized 3-4 Hz spike-wave paroxysms and trains of multiple spikes in the heterozygous patient, and photosensitivity in the heterozygous patient and parent as well as in the sibling with homozygous deficiency. The heterozygous patient and parents did not manifest 4-OH-butyric aciduria but SSADH levels were low and a splice site mutation of the SSADH gene was identified in each. CONCLUSIONS: Heterozygosity for SSADH deficiency may be associated with an epilepsy syndrome characterized by absence and myoclonic seizures, photoparoxysmal EEG and generalized epileptiform discharges SIGNIFICANCE: Heterozygous SSADH deficiency may be suspected, given an appropriate family history in the setting of an apparently idiopathic generalized epilepsy. Pathogenic explanations may relate to regional elevations in GABA or GHB concentrations. PMID- 15134712 TI - Pathological theta oscillations in idiopathic generalised epilepsy. AB - OBJECTIVE: To investigate spectral power, inter- and intra-hemispheric coherence in the interictal scalp electroencephalography (EEG) of 41 patients with idiopathic generalised epilepsy. METHODS: Two minutes of eyes-closed waking interictal EEG activity was analysed. Fast Fourier transformation was performed. Raw and age-regressed, Z-transformed values were computed for 19 electrodes and 4 frequency bands: absolute power (AP, ZAP), percent power (RP, ZRP), band mean frequency (MF, ZMF), inter-hemispheric (CO, ZCO) and intra-hemispheric (IC, ZIC) coherence. Compressed values (scalp averages) were computed for each parameter and 4 frequency bands. Compressed data of the patients (GE group) and the healthy controls (C group) were compared. RESULTS: ZAP across the entire 1.5-25.0 Hz range was greater in the GE than in the C group. Delta and theta ZRP was greater, alpha ZRP was less in GE than in C. ZMF and ZIC was about the same in the GE and C groups. The crucial, band-specific finding was significantly greater theta-ZCO in the GE group, in contrast to normal or decreased ZCO in the other bands. In addition, within-group correlation between ZAP of the frequency bands, correlation of ZAP and ZCO, correlation of ZIC and ZCO were different for the two groups. CONCLUSIONS: The pattern of enhanced ZAP, ZRP, ZCO, together with normal ZIC and ZMF in the theta range was a peculiar, novel finding in GE. It was incompatible with any of the known patterns of altered power and coherence due to lesional or metabolic aetiology. This pattern presumably indicates the presence of a powerful, diffuse, hypersynchronous, hypercoherent theta oscillation at the thalamo-cortical level of the brain. The role of inter-hemispheric connections in maintaining this oscillation was suggested. The other findings suggest disturbed central regulation of EEG power and coherence in the interictal state. PMID- 15134713 TI - Confidence limits of dipole source reconstruction results. AB - OBJECTIVE: Equivalent dipole models are widely used in electro-encephalo-graphic (EEG) and magneto-encephalo-graphic (MEG) source reconstruction. Despite their point-like definition, the best-fit solutions have a certain probability volume depending on the source position and orientation as well as on the actually used sensor set-up and the signal-to-noise ratio (SNR). In order to avoid the misleading impression of exact localization results, a measure of the SD of the dipole localization is desirable. METHODS: This measure can be obtained by performing a deviation scan around the best-fit positions, where the explainable field is determined and compared to the best-fit field. Using a linear approximation, confidence ellipsoids can then be computed and their axes and volumes can be determined by relating the field differences to the noise of the measured data. Test-dipoles inside of a 3 spherical shells volume conductor model were used to simulate EEG- and MEG-data with sources of known positions, orientations, and noise levels. Confidence ellipsoids were computed for these test-dipole solutions and deviation scans around the best-fit dipole positions were performed in order to compare the size and the shape of the confidence ellipsoids with the real error-hypersurface. SDs of repeated dipole localizations at different depths were computed to show the validity of the linear approximation over the whole eccentricity range. RESULTS: The size of the axes of the confidence ellipsoids is inversely proportional to the SNR of the measured data, thus the confidence volume is inversely proportional to the third power of the SNR. Good agreement between SDs of repeated dipole localizations and the confidence ellipsoids was found for both EEG- and MEG-cases. CONCLUSIONS: The new method adds a new and important dimension to dipole source reconstruction results by characterizing their reliability. It is also very helpful in deciding how many dipoles are necessary to explain the measured data, since superfluous dipoles exhibit rather large confidence volumes. PMID- 15134714 TI - Clinical utility of dorsal sural nerve conduction studies in healthy and diabetic children. AB - OBJECTIVE: Monitoring of the dorsal sural sensory nerve action potential (SNAP) is a sensitive method for detection of peripheral neuropathies. We tried to determine the normal dorsal sural nerve conduction values of the childhood population and assessed the clinical utility of this method in diabetic children who have no clinical sign of peripheral neuropathy. METHODS: In the study, 36 healthy and 27 diabetic children were included. In all subjects peripheral motor and sensory nerve studies were performed on the upper and lower limbs including dorsal sural nerve conduction studies. RESULTS: The dorsal sural SNAP mean amplitude was 8.24+/-3.08 microV, mean latency was 2.47+/-0.48 ms, mean sensory conduction velocity was 41.63+/-5.43 m/s in healthy children. Dorsal sural SNAPs were absent bilaterally in one diabetic patient. In the other 26 diabetic patients, the mean dorsal sural nerve distal latency was longer (2.93+/-0.63 ms, P = 0.004), mean SCV was slower than in healthy subjects (36.68+/-7.66 m/s, P = 0.005). However, dorsal sural nerve amplitude was not different between the groups. A dorsal sural nerve latency of more than 2.9 ms had a sensitivity of 50% and a specificity of 75%. A dorsal sural nerve velocity of less than 36 m/s had a sensitivity of 54% and a specificity of 92%. CONCLUSIONS: We designated the reference values of the dorsal sural nerve in healthy children. In addition, our findings suggest that dorsal sural nerve conduction studies may have value to determine neuropathy in the early stages in children with diabetes. SIGNIFICANCE: The dorsal sural nerve conduction studies in diabetic children may have value to determine the neuropathy in its early stages. PMID- 15134715 TI - Muscle membrane polarisation after provocative tests, and after cooling: the normal CMAP changes to be expected. AB - OBJECTIVE: To know the range of changes of compound muscle action potentials (CMAPs) in the muscles innervated by the ulnar nerve after diverse provocative tests, 14 healthy patients were studied with the same protocol. METHODS: CMAPs were measured at rest, just after a short exercise test (SET), during short 5 and 10c/s repetitive nerve stimulation (RNS) trains, at approximately 32 and approximately 20 degrees C. RESULTS: At 32 degrees C, the SET induced a significant but transient enlargement of the CMAPs (amplitude increased by 8.3%, duration decreased by 9%) that was only partially reproduced by RNS trains, except for a significant shortening of the CMAPs at 10c/s. At 20 degrees C without exercise, CMAPs increased significantly by approximately 30% in amplitude, duration and area, and after the SET the inverse of what has been seen at 32 degrees C was observed (amplitude decreased by 1.7% and duration increased by 9%). RNS at 20 degrees C produced a marked interpatient heterogeneity except for a significant shortening of the CMAPs at 10c/s. In one pure autonomic failure patient, the infusion of norepinephrine induced potentiation of the responses at rest and a decrease in the expected changes after provocative tests. CONCLUSIONS: CMAP amplitude and duration are significantly modified just after the SET at 32 degrees C, at rest at 20 degrees C and after RNS at 10c/s but not at 5c/s. Although providing indirect evidence, these findings indicate that provocative tests make the muscle membrane hyperexcitable by the way of a direct influence on the electrical events and by an indirect local catecholamine spillover. PMID- 15134716 TI - Clinical and electrophysiological follow-up after local steroid injection in the carpal tunnel syndrome. AB - OBJECTIVE: Local steroid injections are used for treatment for the carpal tunnel syndrome (CTS). Study of changes in neurophysiological parameters after such treatment for idiopathic CTS might be a supportive argument for the effectiveness of steroid treatment. METHODS: Twenty-one patients with CTS were included and evaluated before and at 1, 3 and 6 months after treatment. At the inclusion date patients received a single 1 ml local steroid injection. Various electrophysiological tests were used. For clinical evaluation, we used the Boston Carpal Tunnel Questionnaire (BCTQ) and the General Outcome Score. RESULTS: The mean nerve conduction parameters had significantly improved at 1 month, except the SNAP4 and CMAP. This improvement was still present at 3 months. At 6 months follow-up, the improvement in the SDL4, SNAP4, M-U, DML and CMAP remained significant. In 6 patients (29%), the nerve conduction parameters normalized and remained so until the end of the study. The BCTQ and the General Outcome Score significantly improved as well. However, there was no correlation between the electrophysiological data, the BCTQ and the General Outcome Score. CONCLUSIONS: The improvements of nerve conduction parameters independently support the ideas on effectiveness of steroid injection therapy in CTS. PMID- 15134717 TI - Transient phrenic nerve palsy after cardiac operation in infants. AB - OBJECTIVE: The aims of this study were to prove the presence of transient phrenic nerve palsy in children after cardiac surgery by successive recordings of diaphragmatic action potentials (DAPs), and to decide the indication of diaphragmatic plication in infants with postoperative phrenic nerve palsy. METHODS: The DAPs were recorded from 11 infants (age 0-54 months) under artificial ventilation after cardiac surgery. The successive DAP recordings were performed within 3-4 days (0W), 1 week (1W) and 2 weeks (2W) after operation to make a final decision for diaphragmatic plication to wean artificial ventilation. RESULTS: The patients were divided into 3 groups according to the DAP changes in successive recordings, namely, patients with normal DAPs at 0W, patients with transient depression of DAPs at 0W followed by recovery to normal DAPs by 1W and/or 2W, and patients with persistent depression of DAPs of the affected side necessitating plication of hemidiaphragm. CONCLUSIONS: In infants with phrenic nerve palsy after cardiothoracic surgery, persistently abnormal DAPs in repeated electrophysiologic examinations for at least 2 weeks after surgery are a useful guidance to support clinical and radiological evidence for an indication of diaphragmatic plication. PMID- 15134718 TI - Cardiovascular autonomic dysfunction correlates with brain MRI lesion load in MS. AB - OBJECTIVE: The aim of the present study was to investigate the cardiovascular autonomic control in clinically definite multiple sclerosis (MS) patients with a standardised battery of cardiovascular tests and to correlate these findings with the brain magnetic resonance imaging (MRI) lesion load. METHODS: Fifty-one patients with MS and 50 healthy controls were studied. Brain MRI was performed in all patients showing typical MS lesions. The cardiovascular tests were carried out using a standardised battery. RESULTS: Heart rate (HR) responses to deep breathing (P < 0.05) and tilt table testing (P < 0.001) were significantly decreased in MS patients when compared to those of the controls. Blood pressure (BP) responses in the tilt table test were also impaired in MS patients (diastolic P < 0.001, systolic P < 0.05). Of the different brain areas investigated the total volume of the midbrain MRI lesions (P < 0.05) was the one most clearly associated with the impaired BP responses. CONCLUSIONS: MS results in both reduced HR variation and decreased BP reactions indicating disturbed cardiovascular regulation. In particular, the midbrain lesions found in MS are associated with cardiovascular dysfunction. PMID- 15134719 TI - Motor cortex stimulation for amyotrophic lateral sclerosis. Time for a therapeutic trial? AB - OBJECTIVE: Repetitive transcranial magnetic stimulation (rTMS) of the brain can modulate neurotransmission. The aim of this preliminary study was to investigate whether rTMS of the motor cortex at low (1 Hz) or high (20 Hz) frequencies can have any beneficial effect in a transgenic rat model of amyotrophic lateral sclerosis (ALS) and in a few patients with ALS. METHODS: The effects of chronic rTMS were evaluated in 20 transgenic rats overexpressing the human G93A mutant superoxide dismutase 1 gene. Several cycles of rTMS were also performed in 4 ALS patients and the rate of progression of the disease before and during rTMS treatment was compared. RESULTS: No effects of rTMS was observed in transgenic rats. The rTMS treatment was well tolerated by the patients. All ALS patients continued to deteriorate. However, in the patients exposed to low-frequency rTMS the rate of progression during treatment was slightly slower than that evaluated before treatment; an opposite tendency was observed in patients exposed to high frequencies. CONCLUSIONS: Though we cannot be sure whether the effects observed in the patients can be attributed to rTMS, further investigation using low frequency motor cortex stimulation on a larger group of ALS patients is warranted. SIGNIFICANCE: The results of the pilot study in humans might open up a new therapeutic perspective in ALS based on neuromodulation. PMID- 15134720 TI - Recent achievements in fundamental and practical aspects of ERP. Proceedings from the VIIIth International Workshop on Electron Magnetic Resonance of Disordered Systems (EMARDIS). Sofia, Bulgaria, 7-16 June 2003. PMID- 15134721 TI - An EPR method for probing surface magnetic fields, dipolar distances, and magnetization fluctuations in single molecule magnets. AB - We outline a spectroscopic method for probing the effective magnetic field B on the surface of crystals of the single molecule magnet (SMM) [(C6H15N3)6Fe8(mu3 O)2(mu2-OH)12]Br7(H2O)Br.H2O, (Fe8Br8). This technique utilizes the line shape changes in the EPR spectra of the organic radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) (g = 2.0036, single peak) adsorbed onto the sample. The temperature dependence of the EPR line shifts scale with the sample's magnetization as measured by a SQUID magnetometer. Analysis of the line shape in terms of dipolar coupling between the DPPH and the SMM molecules on the surface, yields their average dipolar distance. The method's potential for measuring the magnetization fluctuation dynamics is briefly pointed out using the SMM [Mn12O12(CH3COO)16(H2O)4].2CH3COOH.4H2O (Mn12-acetate). PMID- 15134722 TI - EPR and NMR in powders of doped and undoped IV-VI crystals. AB - The results of the extensive investigations of the variation of the EPR and NMR spectra of active centers due to the existence of the native defects generated by disorder in the IV-VI semiconductor matrices are presented. Both undoped and doped with Gd impurity powder samples of different grain sizes, made from Pb(1 x)Sn(x)Te crystals with the composition in the range 0 < or = x < or = 0.2 were studied. Impurity Gd ions were used as the paramagnetic EPR probe, whereas the 207 Pb nuclei as the NMR probe. The following aspects have been ascertained. (i) Grinding of the initial single crystals into powders leads to an additional component line appearing in the NMR spectra of the 207 Pb nuclei and also to a significant increase in the intensity of EPR spectra of the impurity Gd ions. (ii) Both the Gd EPR spectra as well as the 207 Pb NMR spectra undergo modifications due to isothermal annealing, whereas the character of these modifications is determined by both the temperature and duration of the thermal treatment applied. (iii) Some characteristic correlation between the variation of the EPR spectra of impurity Gd ions and that of the NMR spectra of 207 Pb nuclei, which results from the annealing of the samples, has been observed. Experimental results are interpreted based on the prevailing models of the behavior of the doped impurities and the native defects in the lead and tin telluride crystals. PMID- 15134723 TI - Paramagnetic species on catalytic surfaces--DFT investigations into structure sensitivity of the hyperfine coupling constants. AB - Structure sensitivity of the hyperfine coupling constants was investigated by means of DFT calculations for selected surface paramagnetic species. A *CH2OH radical trapped on silica and intrazeolite copper nitrosyl adducts encaged in ZSM 5 were taken as the examples. The surface of amorphous silica was modeled with a [Si5O8H10] cluster, whereas the zeolite hosting sites were epitomized by [Si4AlO5(OH)10]- cluster. Three different coordination modes of the *CH2OH radical were considered and the isotropic 13C and 1H hyperfine constants of the resultant van der Waals complexes, calculated with B3LYP/6-311G(d), were discussed in terms of the angular deformations caused by hydrogen bonds with the cluster. The magnetic parameters of the eta1-N[CuNO]11 and eta1-O[CuNO]11 linkage isomers were calculated at the BPW91/LanL2DZ and 6-311G(df) level. For the most stable eta1-N adduct a clear dependence of the spin density distribution within the Cu-NO moiety on changes in the Cu-N-O angle and the Cu-N bond distance was observed and accounted for by varying spin polarization and delocalization contributions. PMID- 15134724 TI - Nitric oxide adsorbed on zeolites: EPR studies. AB - CW-EPR studies of NO adsorbed on sodium ion-exchanged zeolites were focused on the geometrical structure of NO monoradical and (NO)2 biradical formed on zeolites. The EPR spectrum of NO monoradical adsorbed on zeolite can be characterized by the three different g-tensor components and the resolved y component hyperfine coupling with the 14N nucleus. Among the g-tensor components, the value of g(zz) is very sensitive to the local environment of zeolite and becomes a measure of the electrostatic field in zeolite. The temperature dependence of the g-tensor demonstrated the presence of two states of the Na-NO adduct, in rigid and rotational states. The EPR spectra of NO adsorbed on alkaline metal ion-exchanged zeolite and their temperature dependency are essentially the same as that on sodium ion-exchanged zeolite. On the other hand, for NO adsorbed on copper ion-exchanged zeolite it is known that the magnetic interaction between NO molecule and paramagnetic copper ion are observable in the spectra recorded at low temperature. The signals assigned to (NO)2 biradical were detected for EPR spectrum of NO adsorbed on Na-LTA. CW-EPR spectra as well as their theoretical calculation suggested that the two NO molecules are aligned along their N-O bond axes. A new procedure for automatical EPR simulation is described which makes it possible to analyze EPR spectrum easily. In the last part of this paper, some instances when other nitrogen oxides were used as a probe molecule to characterize the zeolite structure, chemical properties of zeolites, and dynamics of small molecules were described on the basis of selected literature data reported recently. PMID- 15134725 TI - Chromophoric spin-labeled beta-lactam antibiotics for ENDOR structural characterization of reaction intermediates of class A and class C beta lactamases. AB - The chromophoric spin-label substrate 6-N-[3-(2,2,5,5-tetramethyl-1-oxypyrrolin-3 yl)-propen-2-oyl]penicillanic acid (SLPPEN) was synthesized by acylation of 6 aminopenicillanic acid with the acid chloride of 3-(2,2,5,5-tetramethyl-1 oxypyrrolinyl)-2-propenoic acid and characterized by physical methods. By application of angle-selected electron nuclear double resonance (ENDOR), we have determined the molecular structure of SLPPEN in solution. SLPPEN exhibited UV absorption properties that allowed accurate monitoring of the kinetics of its enzyme-catalyzed hydrolysis. The maximum value of the (substrate-product) difference extinction coefficient was 2824 M(-1) cm(-1) at 275 nm compared to 670 M(-1) cm(-1) at 232 nm for SLPEN [J. Am. Chem. Soc. 117 (1995) 6739]. For SLPPEN, the steady-state kinetic parameters kcat and kcat/KM, determined under initial velocity conditions, were 637 +/- 36 s(-1) and 13.8 +/- 1.4 x 10(6) M(-1) s(-1), respectively, for hydrolysis catalyzed by TEM-1 beta-lactamase of E. coli, and 0.5 +/- 0.04 s(-1) and 3.9 +/- 0.4 x 10(4) M(-1) s(-1) for hydrolysis catalyzed by the beta-lactamase of Enterobacter cloacae P99. We have also observed "burst kinetics" for the hydrolysis of SLPPEN with P99 beta-lactamase, indicative of formation of an acylenzyme reaction intermediate. In DMSO:H2O (30:70, v:v) cryosolvent mixtures buffered to pH* 7.0, the half-life of the acylenzyme intermediate formed with the P99 enzyme at -5 degrees C was > or = 3 min, suitable for optical characterization. The observation of burst kinetics in the hydrolysis of SLPPEN catalyzed by P99 beta-lactamase suggests that this chromophoric spin-labeled substrate is differentially sensitive to active site interactions underlying the cephalosporinase and penicillinase reactivity of this class C enzyme. PMID- 15134726 TI - Charge-transfer photolysis of copper(II) dithiocarbamate mixed-ligand complexes in toluene/alcohol solutions. AB - Charge-transfer (CT)-photolysis of Cu(II) dithiocarbamate mixed-ligand complexes Cu(II)(Et2dtc)X (X = Cl-, Br-) and Cu(II)(Et2dtc)(+)...Y- (Y = ClO4-, NO3-) has been studied in toluene/ROH and compared with our previous data obtained in chloromethane/ROH solutions, where chloromethane = CCl4, CHCl3 or CH2Cl2 and ROH = MeOH, EtOH, i-PrOH or i-BuOH. An EPR evidence is obtained about the formation of a new copper(II) dithiocarbamate mixed-ligand complex during simultaneous photolyses of Cu(II)(Et2dtc)+ and Cu(II)(Et2dtc)2 species in toluene/ROH. The role of the solvent is discussed from the combined analysis of spectrophotometric and EPR data and quantum yield results. PMID- 15134727 TI - Structure of copper(II) dithiocarbamate mixed-ligand complexes and their photoreactivities in alcohols. AB - The composition of the co-ordination sphere of Cu(II) dithiocarbamate mixed ligand complexes Cu(Et2)dtc)X (X = Cl-, Br-) and Cu(Et2)dtc)(+)...Y- (Y- = ClO4-, NO3-) is studied from the combined analysis of spectrophotometric and EPR data. The results obtained about CT-photolysis of the complexes in EtOH and i-PrOH are compared with our previous data of photolysis in chloromethane/ROH solutions. Reaction mechanism and the role of alcohol are discussed on the ground of electronic and EPR spectra and quantum yield results. PMID- 15134728 TI - EPR and UV spectral study of gamma-irradiated white and burned sugar, fructose and glucose. AB - The EPR and UV spectral properties of gamma-irradiated white and burned sugar, fructose and glucose are studied with the accent on their suitability as dosimetric materials. It is shown that gamma-irradiation of solid samples of white sugar and fructose yields stable EPR spectra whereas glucose signal remains time-dependent even 11 months later. Sugar and glucose exhibit linear EPR dose response in the region 0.44-21 kGy and fructose only up to ca. 10 kGy. The relative radiation sensitivity obtained for sugar and fructose is up to 10 kGy and slightly lower for glucose. Burned saccharides provide 2-3 orders of magnitude lower EPR radiation sensitivity making them not suitable for the proposed designation. According to the UV spectra water solutions of gamma irradiated solid white saccharides show well pronounced absorption bands at 267 and 286 nm for sugar and fructose with time-dependent intensities reaching steady values ca. 11 days after dissolution. The intensities of these absorption bands are in linear relation with the absorbed dose of gamma-radiation. Glucose shows low sensitively because irradiation with 5.5 kGy yields only a shoulder at about 260-280 nm with decreasing to ca. 40% intensity in the first few days after dissolution. Excellent correlation between the intensities of the EPR- and UV absorbed dose response is found for sugar and fructose in the region 0.44-10 kGy. This opens new possibilities for independent calibration the EPR dose response. Finally, the comparison suggests sugar as the best, universal material for EPR- and/or UV-dosimetry in the region 0.44-160 kGy. PMID- 15134729 TI - ESR investigation of sucrose radicals produced by particle irradiation. AB - We investigated sucrose radicals produced by heavy-ion irradiation with various linear energy transfer (LETs) and the possibility for a sucrose electron spin resonance (ESR) dosimeter. The impact of heavy ions on sucrose produced sucrose radicals, which were measured by ESR. The obtained spectral pattern was the same as that for helium (He) ions, carbon (C) ions, neon (Ne) ions, iron (Fe) ions, and gamma-ray irradiation. Identical spectra were measured after 1 year, but the initial intensities decreased by a few percent when the samples were kept in ESR tubes with the caps at ambient temperature. The total spin concentration obtained by heavy-ion irradiation had a linear relation with the absorbed dose, and correlated logarithmically with the LET. Qualitative ESR analyses showed that the production of sucrose radicals depended on both the particle identity and the LET at the same dose. The production of spin concentration by He ions was the most sensitive to LET. Empirical relations between the LET and the spin yield for various particles imply that the LET at a certain dose can be estimated by the spin concentration. Therefore, the present ESR results imply that sucrose can be used to monitor the absorbed dose and the LET of particle irradiation. PMID- 15134730 TI - Compounds of 6Li and natural Li for EPR dosimetry in photon/neutron mixed radiation fields. AB - Formates and dithionates of 6Li, enriched and 7Li in natural composition of Li offer a possibility to measure the absorbed dose from photons and thermal neutrons in a mixed radiation field for instance at a boron neutron capture therapy (BNCT) facility. Tests with formates and dithionates of enriched 6Li and lithium compounds with natural composition have been performed at the BNCT facility at Studsvik, Sweden. Irradiations have been performed at 3 cm depth in a Perspex phantom in a fluence rate of thermal neutrons 1.8 x 10(9) n cm(-2) s(-1). The compounds were also irradiated in a pure X-ray field from a 4MV linear accelerator at 5 cm depth in a phantom with accurately determined absorbed doses. The signal intensity and shape was investigated within 3 h after the irradiation. A single line spectrum attributed to the CO2- radical was observed after irradiation of lithium formate. An increase in line width occurring after neutron irradiation in comparison with photon irradiation of the 6Li sample was attributed to dipolar broadening between CO2- radicals trapped in the tracks of the alpha particles. A spectrum due to the SO3- radical anion was observed after irradiation of lithium dithionate. The signal amplitude increased using the 6Li in place of the Li with natural composition of isotopes, in studies with low energy X-ray irradiation. Due to the decreased line width, caused by the difference in g(N) and I between the isotopes, the sensitivity with 6Li dithionate may be enhanced by an order of magnitude compared to alanine dosimetry. After comprehensive examination of the different combinations of compounds with different amounts of 6Li and 7Li regarding dosimetry, radiation chemistry and EPR properties these dosimeter material might be used for dose determinations at BNCT treatments and for biomedical experiments. Interesting properties of the radical formation might be visible due to the large difference in ionization density of neutrons compared to photons. PMID- 15134731 TI - EPR study of light illumination effects on radicals in gamma-irradiated L alanine. AB - Exposure of gamma-irradiated L-alanine samples to sunlight and to light from a regular, fluorescent lamp resulted in significant changes in their EPR resonance patterns, both to spectral shapes and intensities. The experimental EPR spectra were numerically decomposed into three components reflecting contributions of three different radicals (R1-R3) generated by ionizing radiation in alanine. The light exposure caused a decay of the measured EPR signal intensity. For similar light intensities and exposure times the decay was much more pronounced in samples illuminated by sunlight than in samples illuminated by the fluorescent lamp. In both cases light-induced decay of R1 radicals was observed. Sunlight illumination resulted in a moderate decay of R2 radicals and in a doubling of the R3 radical population. On the other hand, fluorescent light caused a significant increase of R2 radicals and did not change the amount of R3 radicals. A quantitative analysis of the variations of the three radical contributions to the total EPR spectra upon fluorescent light exposure suggests a net R1-->R2 free radical transformation. These effects of light on the alanine dosimetric signal should be taken into account in dosimetry protocols, assuring protection of alanine dosimeters from extended exposure to light. PMID- 15134732 TI - EPR study of gamma induced radicals in amino acid powders. AB - To better understand the composite character of amino acids EPR spectra, the radiolysis and reactions which occurred after irradiation of amino acids, a comparative EPR study of a few simple amino acids has been made in order to identify qualitatively and quantitatively the different radiation-induced radicals in amino acid powders. A spin-trapping methodology has been developed and carried out on irradiated glycine, alanine and valine. PMID- 15134733 TI - Spectroscopic study of bituminous oxidative stress. AB - Bitumen, as each organic substance, is a product which alters over time. Indeed, roads deteriorate under the effect of several phenomena. A number of studies have been undertaken to increase the quality of road's coating, mostly by adding polymer to bitumen. This work was based on the study, by electron paramagnetic resonance (EPR), FTIR and Synchronous UV fluorescence, of different base and modified bitumens after different treatments used to simulate the ageing (gamma irradiation, thermal treatment). Our purpose was to compare and correlate the results obtained by different techniques to improve the knowledge of bitumen's reactivity and evolution submitted to ageing phenomena. PMID- 15134734 TI - Search for EPR markers of the history and origin of the insoluble organic matter in extraterrestrial and terrestrial rocks. AB - The insoluble organic matter (IOM) of three carbonaceous meteorites (Orgueil, Murchison and Tagish Lake meteorites) and three samples of cherts (microcrystalline SiO2 rock) containing microfossils with age ranging between 45 million years and 3.5 billion years is studied by electron paramagnetic resonance (EPR). The age of the meteorites is that of the solar system (4.6 billion years). The purpose of this work was to determine the EPR parameters, which allow us to discriminate between biogenic and extra terrestrial origin for the organic matter. Such indicators should be relevant for the controversy regarding the biogenicity of the organic matter in the oldest cheroot (3.5 billion years) and in Martian meteorites containing microbe-like microstructures. The organic matter of meteorites contains a high concentration of diradicaloid moieties characterised by a diamagnetic ground state S = 0 and a thermally accessible triplet state S = 1. The three meteorites exhibit the same singlet-triplet gap (ST gap) DeltaE approximately 0.1 eV. To the best of our knowledge, such diradicaloids are unknown in insoluble organic matter of terrestrial origin. We have also shown that the EPR linewidth of insoluble organic matter in cherts and coals decrease logarithmically with the age of the organic matter. We conclude from this result that the organic matter in the oldest cherts (3.5 billion years) has the same age as their SiO2 matrix, and is not due to a latter contamination by bacteria, as was recently found in meteoritic samples. PMID- 15134735 TI - ESR studies of Anatolian gypsum. AB - Electron spin resonance (ESR) was used to date the formation and most recent recrystallization of three types of gypsum samples (massive, bedded and fracture filling gypsum) from the Sakarya Formation at Eskisehir in the middle of Turkey. The ESR spectra had the signals of Fe3+ and Mn2+ in addition to those of the G1 and G2 sensitive centers (g = 2.002 and 2.008) to artificial gamma-irradiation. ESR intensities of G1 and G2 were enhanced by gamma-ray irradiation to give equivalent doses DE) for each sample. The ESR ages derived from the annual doses of 238U, 232Th and 40K contents of the samples determined by thermal neutron activation analysis (TNAA) were between 43 +/- 18 ka (massive gypsum) and 1100 +/ 466 ka (bedded gypsum) as minimal and maximal values. The ages fall into the upper Miocene-Pliocene Epoch of the geological time scale, which agreed with the stratigraphy. PMID- 15134736 TI - Selective estimation of soot in home dust by EPR spectrometry. AB - The content of soot in home dust collected from eight different flats or homes situated in different areas of Sofia, Bulgaria, is studied for a period of 14 months by using EPR spectrometry. The results obtained show two different areas in respect to the in-door pollution with soot which in one area is three to five times lower in comparison to the another. The characteristic features of both places are the intensive automotive traffic and central heating in the first part and less intensive automotive traffic but local heating facilities in the second area. These results are fully coinciding with previous studies on the soot content in urban aerosols and strongly suggest that main source of soot in air (in form of aerosols or precipitated as home dust) are the local heating facilities. PMID- 15134737 TI - ESR investigation of the oxidative damage in lungs caused by asbestos and air pollution particles. AB - Exposure to asbestos and air pollution particles can be associated with increased human morbidity and mortality. However, the molecular mechanism of lung injuries remains unknown. It has been postulated that the in vivo toxicity results from the catalysis of free radical generation. Using electron spin resonance (ESR) in conjunction with the spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone (4 POBN) we previously investigated in vivo free radical production by rats treated with intratracheal instillation of asbestos (crocidolite fibers) and an emission source air pollution particle (oil fly ash). In this report we compare the effect of two different exposures on the type of free radicals they induce in in vivo animal model. Twenty-four hours after the exposure, ESR spectroscopy of the chloroform extract from lungs of animals exposed to either asbestos or oil fly ash gave a spectrum consistent with a carbon-centered radical adduct (aN = 15.01 G and aH = 2.46 G). To test whether free radical formation occurred in vivo and not in vitro, a number of control experiments were performed. Combinations (both individually and together) of asbestos or oil fly ash and 4-POBN were added to lung homogenate of unexposed rats prior to chloroform extraction. No detectable ESR signal resulted. To exclude the possibility of ex vivo free radical generation, asbestos or oil fly ash was added to lung homogenate of an animal treated with 4-POBN. Also, 4-POBN was added to lung homogenate from rats instilled with asbestos or oil fly ash. Neither system produced radical adducts, indicating that the ESR signal detected in the lung extracts of the treated animals must be produced in vivo and not ex vivo or in vitro. In conclusion, ESR analysis of lung tissue demonstrated that both exposures produce lipid-derived radical metabolites despite their different composition and structure. Analogously, both exposures provide evidence of in vivo enhanced lipid peroxidation. Furthermore, it is concluded that without the presence of a spin trapping agent, no free radical metabolites could be detected directly by ESR in either exposure. PMID- 15134738 TI - EPR oximetry of tumors in vivo in cancer therapy. AB - The partial oxygen pressure (pO2) in tumors is considered to be one of important factors that affect the response of tumors to different treatment. Therefore, we anticipate that the information about the variation of oxygen concentration in tumors can be used as a guide for individualizing radiotherapy, chemotherapy, and especially the combined therapies. There is thus a need to obtain quantitative data on the effects of different therapies on tumor oxygenation under in vivo conditions. One of the methods, which enable these measurements is EPR oximetry. In this work basic principles of the method will be described as well as some examples of tumor oxygenation changes after application of chemotherapeutic drugs (vinblastine, cisplatin, bleomycin) or electric pulses in combination with cisplatin or bleomycin to fibrosarcoma SA-1 tumors in mice. A paramagnetic probe, a char of Bubinga tree, was implanted into the tumor (center and periphery) and in the muscle or subcutis. EPR spectra line-width, which is proportional to oxygen concentration, was measured with time after the treatments. Tumor oxygenation was reduced for 58% of pretreatment value 1 h after intraperitoneal injection of 2.5 mg kg(-1) VLB and returned to pretreatment level within 24 h. Reduction in oxygenation of muscle and subcutis was much smaller and returned to pretreatment value faster as in tumors. With cisplatin (4 mg kg(-1)) and bleomicyn (1 mg kg(-1)) the reduction was less than 15%, but increases in combined therapy to 70%. Similar reduction was observed also with electric pulses alone (eight pulses, 1300 V cm(-1), 100 micros, 1 Hz) with fast recovery of 8h. After electrochemotherapy the recovery was slower and occurs only after 48 h. This study demonstrates that EPR oximetry is a sensitive method for monitoring changes in tissue oxygenation after different treatments, which may have implications in controlling side effects of therapy and in the planning of combined treatments. PMID- 15134739 TI - Organic matter transformation in the environment investigated by quantitative electron paramagnetic resonance (EPR) spectroscopy: studies on lignins. AB - The lignins separated from angiosperm and gymnosperm trees, peat and xylitic brown coal were investigated by quantitative EPR. Observed free radicals in lignins are sensitive to alkaline environment. Gaseous ammonia interacting with solid lignins in resonance cavity shifts quinone-hydroquinone equilibria towards formation of semiquinone anions. Complexation of copper(II) by lignins causes drastic decrease of the semiquinones in the matrices. Formation of lignin-Pb(II) complexes yielded radicals characterised by unusually low g-value (1.9999 2.0003). Monomeric structural units of the investigated lignins were recognised by pyrolysis with in situ methylation by tetramethylammonium hydroxide. Although for the natural lignins the mixture of normal semiquinone signals at g about 2.0034 and signals at g 1.9999 were observed, some monomeric components of lignins (e.g., caffeic acid, pyrogallol) gave pure lines at g = 1.9999. The bacterial oxidative biodegradation of lignin monomeric components and their Pb(II) complexes resulted in increase of the radical signals. PMID- 15134740 TI - EPR study of free radicals in bread. AB - The features of the recorded EPR spectra of paramagnetic species formed in bread and rusk are reported. The appearance of free radicals in them is only connected with their thermal treatment since the starting materials (flour and grains) exhibit very weak EPR signal. The obtained EPR spectra are complex and indicate that: (i) the relative number of paramagnetic species depends on the temperature and treating time of the raw product; (ii) the g-values are strongly temperature dependent with a tendency to coincide at t > or = 220 degrees C. Because of the relatively low (150-220 degrees C) temperature of thermal treatment, the studied free radicals can be assumed to appear in the course of the browning (Maillard) reaction and not to the carbonization of the material. PMID- 15134741 TI - A determination of antioxidant efficiencies using ESR and computational methods. AB - Using Transition-State Theory, experimental rate constants, determined over a range of temperatures, for reactions of Vitamin E type antioxidants are analysed in terms of their enthalpies and entropies of activation. It is further shown that computational methods may be employed to calculate enthalpies and entropies, and hence Gibbs free energies, for the overall reactions. Within the linear free energy relationship (LFER) assumption, that the Gibbs free energy of activation is proportional to the overall Gibbs free energy change for the reaction, it is possible to rationalise, and even to predict, the relative contributions of enthalpy and entropy for reactions of interest, involving potential antioxidants. A method is devised, involving a competitive reaction between *CH3 radicals and both the spin-trap PBN and the antioxidant, which enables the relatively rapid determination of a relative ordering of activities for a series of potential antioxidant compounds, and also of their rate constants for scavenging *CH3 radicals (relative to the rate constant for addition of *CH3 to PBN). PMID- 15134743 TI - Sucrose metabolism: regulatory mechanisms and pivotal roles in sugar sensing and plant development. AB - Sucrose cleavage is vital to multicellular plants, not only for the allocation of crucial carbon resources but also for the initiation of hexose-based sugar signals in importing structures. Only the invertase and reversible sucrose synthase reactions catalyze known paths of sucrose breakdown in vivo. The regulation of these reactions and its consequences has therefore become a central issue in plant carbon metabolism. Primary mechanisms for this regulation involve the capacity of invertases to alter sugar signals by producing glucose rather than UDPglucose, and thus also two-fold more hexoses than are produced by sucrose synthase. In addition, vacuolar sites of cleavage by invertases could allow temporal control via compartmentalization. In addition, members of the gene families encoding either invertases or sucrose synthases respond at transcriptional and posttranscriptional levels to diverse environmental signals, including endogenous changes that reflect their own action (e.g. hexoses and hexose-responsive hormone systems such as abscisic acid [ABA] signaling). At the enzyme level, sucrose synthases can be regulated by rapid changes in sub-cellular localization, phosphorylation, and carefully modulated protein turnover. In addition to transcriptional control, invertase action can also be regulated at the enzyme level by highly localized inhibitor proteins and by a system that has the potential to initiate and terminate invertase activity in vacuoles. The extent, path, and site of sucrose metabolism are thus highly responsive to both internal and external environmental signals and can, in turn, dramatically alter development and stress acclimation. PMID- 15134744 TI - Solute transporters as connecting elements between cytosol and plastid stroma. AB - Metabolite translocators in the inner membrane of the plastid envelope are the interface between cytosolic and plastidial metabolism. Hence, they integrate plastidial pathways, such as photosynthesis, starch biosynthesis, the oxidative pentose phosphate pathway and the shikimate pathway, into the metabolic network of plant cells. Metabolite transporters not only catalyze the flux of metabolites between compartments but also represent information pathways that communicate the metabolic status of the various compartments within plant cells. Recently, a pentose-phosphate translocator was shown to be a novel member of the phosphate translocator protein family. Furthermore, a protein of previously unknown function was identified as a novel type of maltose transporter, and a glutamate/malate translocator that is involved in photorespiration was discovered. In addition, the pathway for maltose metabolism in the cytosol has been unraveled. PMID- 15134745 TI - Respiratory metabolism: glycolysis, the TCA cycle and mitochondrial electron transport. AB - The respiratory pathways of glycolysis, the tricarboxylic acid (TCA) cycle and the mitochondrial electron transport chain are ubiquitous throughout nature. They are essential for both energy provision in heterotrophic cells and a wide range of other physiological functions. Although the series of enzymes and proteins that participate in these pathways have long been known, their regulation and control are much less well understood. Further complexity arises due to the extensive interaction among these pathways in particular, and also between cytosolic and mitochondrial metabolism in general. These interactions include those between mitochondrial function in the photosynthetic and photorespiratory processes, amino-acid biosynthesis and the regulation of cellular redox. Recently, a wide range of molecular and biochemical strategies have been adopted to elucidate the functional significance of these interactions. PMID- 15134746 TI - Green light for galactolipid trafficking. AB - Galactolipids not only play a crucial role in photosynthesis but are also important for the adaptation of membrane-lipid composition in plants to phosphate limiting conditions. The enzymes of galactolipid assembly have been localised to the envelope membranes of chloroplasts. Lipid trafficking is essential for galactolipid synthesis and redistribution because lipid precursors originate from two compartments, the endoplasmic reticulum (ER) and the plastid, and because galactolipids have to be transported to extraplastidial membranes during phosphate deprivation. Analysis of Arabidopsis mutants that are impaired in galactolipid synthesis (i.e. dgd1 and dgd2) or in ER-to-plastid lipid transport (i.e. tgd1) has resulted in the identification of a processive galactosyltransferase whose function is still enigmatic. PMID- 15134747 TI - Phosphatidylglycerol and sulfoquinovosyldiacylglycerol: anionic membrane lipids and phosphate regulation. AB - Photosynthetic membranes of organisms from cyanobacteria to seed plants are characterized by the neutral galactolipids and the anionic glycerolipids sulfoquinovosyldiacylglycerol and phosphatidylglycerol. Recent findings have brought new insights into the biosynthesis of the anionic membrane lipids, the evolutionary origin of the enzymes involved in this process, and the importance of phosphatidylglycerol and sulfoquinovosyldiacylgycerol in photosynthesis. Photosynthetic membranes require a defined level of anionic membrane lipids for proper function, and phosphatidylglycerol and sulfoquinovosyldiacylglycerol can substitute for each other to a certain extent. A defined level of phosphatidylglycerol is, however, indispensable for photoautotrophic growth. On the other hand, sulfoquinovosyldiacylglycerol plays a conditionally important role in enabling photosynthetic organisms to survive the phosphate-limiting conditions frequently encountered in natural habitats. PMID- 15134748 TI - Nucleotide sugar interconversions and cell wall biosynthesis: how to bring the inside to the outside. AB - Plants possess a sophisticated sugar biosynthetic machinery comprising families of nucleotide sugar interconversion enzymes. Literature published in the past two years has made a major contribution to our knowledge of the enzymes and genes involved in the interconversion of nucleotide sugars that are required for cell wall biosynthesis, including UDP-L-rhamnose, UDP-D-galactose, UDP-D-glucuronic acid, UDP-D-xylose, UDP-D-apiose, UDP-L-arabinose, GDP-L-fucose and GDP-L galactose. Indirect evidence suggests that enzyme activity is crudely regulated at the transcriptional level in a cell-type and differentiation-dependent manner. However, feedback inhibition and NAD(+)/NADH redox control, as well as the formation of complexes between differentially encoded isoforms and glycosyltransferases, might fine-tune cell wall matrix biosynthesis. I hypothesise that the control of nucleotide sugar interconversion enzymes regulates glycosylation patterns in response to developmental, metabolic and stress-related stimuli, thereby linking signalling with primary metabolism and the dynamics of the extracellular matrix. PMID- 15134749 TI - Glycosyltransferases and cell wall biosynthesis: novel players and insights. AB - Plants need an enormous biosynthetic machinery to synthesize the complex polysaccharides that are present in the plant cell wall. The isolation, characterization and mapping of wall mutants, together with biochemical approaches, have led to significant advances in our understanding of both wall polysaccharide synthesis at a molecular level and the function of polysaccharides in plant growth and development. Moreover, potential regulation mechanisms and associated protein factors are emerging from recent data. PMID- 15134750 TI - The plot thickens: New perspectives of primary cell wall modification. AB - Recent studies have further confirmed the ubiquity of cell wall restructuring during plant growth and development, and have emphasized the fact that our understanding of the breadth of molecular processes that mediate wall modification is still rudimentary. In the past few years, both enzymatic and non enzymatic agents that apparently contribute to wall disassembly have been identified, and it is likely that additional mechanisms will continue to be revealed. These discoveries are being propelled by the development of new biochemical and biophysical assays, by database mining in the wake of the explosion of plant sequence information from genome sequencing and expressed sequence tags, and by a variety of strategies used to catalog the cell wall proteome. The daunting question of how these mechanistically diverse and complex processes are coordinated remains unresolved. PMID- 15134751 TI - Control of storage-product synthesis in seeds. AB - Seeds provide humans with much of their diet and have been targets for improvement for millennia. The recent development of a range of methodologies for investigating the control of seed metabolism will allow rapid progress towards understanding this process in the future. In situ measurements of metabolite concentrations, in combination with the localisation of gene expression, in developing legume seeds have led to the description of detailed models of the control of starch and protein synthesis. In oilseeds, the application of recently developed 13C-labelling methods allows the quantification of carbon fluxes through individual pathways in the cytosol and plastid. Molecular and genetic approaches are being used in combination to probe both the importance of individual steps in the pathways of storage-product synthesis and potential regulators of the entire process. PMID- 15134752 TI - Understanding flux in plant metabolic networks. AB - The revolutionary growth in our ability to identify the 'parts list' of cellular infrastructure in plants in detail, and to alter it with precision, challenges us to develop methods to quantify how these parts function. For components of metabolism, this means mapping fluxes at the level of metabolic networks. Advances in experimental, analytical and software tools for metabolic flux analysis now allow maps of the fluxes through central metabolism to be obtained from the results of stable-isotope-labeling experiments. Such maps have led to notable successes in understanding and engineering metabolic function in microorganisms. Recent studies in plants are giving insight into particular fluxes, such as those of the pentose phosphate pathway, and into general phenomena, such as substrate- or futile-cycles and compartmentation. The importance of experimental design and statistical analysis have been illustrated, and analyses of fluxes in heterotrophic plant tissues have been carried out recently. PMID- 15134753 TI - Numerous posttranslational modifications provide opportunities for the intricate regulation of metabolic enzymes at multiple levels. AB - The metabolic plasticity displayed by plants during normal development, and in response to environmental fluctuations and stressors, is essential for their growth and survival. The capacity to regulate metabolic enzymes intricately arises in part from posttranslational modifications that can affect enzymatic activity, intracellular localization, protein-protein interactions, and stability. Protein phosphorylation and thiol/disulfide redox modulation are important modifications in plants, and it is likely that O-glycosylation and S nitrosylation will also emerge as important mechanisms. Recent advances in the field of proteomics, in particular the development of novel and specific chemistries for the detection of a diverse number of modifications, are rapidly expanding our awareness of possible modifications and our understanding of the enzymes whose functions are likely to be regulated posttranslationally. PMID- 15134754 TI - Reactive oxygen signalling: the latest news. AB - During the past two years, a wide range of plant responses have been found to be triggered by hydrogen peroxide that is generated in a genetically controlled manner by NADPH oxidases. Several studies have revealed examples of how changes in the concentrations of reactive oxygen species (ROS) are perceived and transferred into signals that change the transcription of genes. Moreover, both the chemical identity of a given ROS and the intracellular site of its production seem to affect the specificity of its biological activity, further increasing the complexity of ROS signalling within plants. PMID- 15134755 TI - Lipid signaling. AB - Various lipids are involved in mediating plant growth, development and responses to biotic and abiotic cues, and their production is regulated by lipid-signaling enzymes. Lipid-hydrolyzing enzymes play a pivotal role both in the production of lipid messengers and in other processes, such as cytoskeletal rearrangement, membrane trafficking, and degradation. Studies on the downstream targets and modes of action of lipid signals in plants are still in their early stages but distinguishing features of plant lipid-based signaling are being recognized. Phospholipase D enzymes and phosphatidic acid may play a broader role in lipid signaling in plants than in other systems. PMID- 15134756 TI - Lipid species profiling: a high-throughput approach to identify lipid compositional changes and determine the function of genes involved in lipid metabolism and signaling. AB - The development of electrospray ionization mass spectrometry has provided the foundation for the development of strategies to identify and quantify complex lipids from unfractionated extracts of small biological samples. In the 1990s, the feasibility of detailed lipid profiling was demonstrated; in the past two years, analytical strategies have been extended to include classes of lipids that are unique to plants. High-throughput lipid profiling by electrospray ionization tandem mass spectrometry, in combination with forward- or reverse-genetics approaches, has recently been utilized to identify lipid metabolic pathways that are involved in plant development and stress responses, to specify the roles of particular genes and enzymes in plant responses to environmental cues, to determine the lipid species that serve as the substrates and products of specific enzymes, and to identify lipid-metabolizing enzymes that are involved in varied plant processes. PMID- 15134757 TI - Minimally invasive dynamic imaging of ions and metabolites in living cells. AB - By 2010, it is expected that biochemical functions will be assigned to many of the products of the approximately 30,000 Arabidopsis genes. Moreover, systematic analysis of mutants will provide insight into the biological function of the gene products. Metabolomic technologies complement these approaches by testing for changes in cellular ion and metabolite patterns, providing essential information for the construction of cellular and whole-plant models of metabolism. However, one important set of information that is especially relevant for multicellular organisms is still lacking, that is, knowledge of the cellular and subcellular variation in metabolite levels. The recent development of protein-based nanosensors for metabolites will help to close this gap by providing a set of tools that can be used to determine cytosolic and subcellular metabolite levels in real time using fluorescence-based microscopy. A major challenge for the future is the application of these nanosensors to quantify metabolite levels in plant cells and tissues. PMID- 15134758 TI - Consciousness, art, and the brain: lessons from Marcel Proust. AB - In his novel Remembrance of Things Past, Marcel Proust argues that conventional descriptions of the phenomenology of consciousness are incomplete because they focus too much on the highly-salient sensory information that dominates each moment of awareness and ignore the network of associations that lies in the background. In this paper, I explicate Proust's theory of conscious experience and show how it leads him directly to a theory of aesthetic perception. Proust's division of awareness into two components roughly corresponds to William James' division of the stream of thought into a "nucleus" and "fringe." Proust argues that the function of art is to evoke the underlying associative network indirectly in the mind of the observer by using carefully chosen sensory surfaces to control the stream of thought. I propose a possible neural basis for this Proustian/Jamesian phenomenology, and argue that the general principles of Proustian aesthetics can be applied to all forms of art. I conclude that a scientific theory of art should follow in a straightforward manner from a scientific theory of consciousness. PMID- 15134759 TI - Aesthetic experience: Marcel Proust and the neo-Jamesian structure of awareness. PMID- 15134760 TI - Memory, imagination, and the cognitive value of the arts. PMID- 15134761 TI - Criteria for an effective theory of consciousness and some preliminary attempts. AB - In the physical sciences a rigorous theory is a hierarchy of descriptions in which causal relationships between many general types of entity at a phenomenological level can be derived from causal relationships between smaller numbers of simpler entities at more detailed levels. The hierarchy of descriptions resembles the modular hierarchy created in electronic systems in order to be able to modify a complex functionality without excessive side effects. Such a hierarchy would make it possible to establish a rigorous scientific theory of consciousness. The causal relationships implicit in definitions of access consciousness and phenomenal consciousness are made explicit, and the corresponding causal relationships at the more detailed levels of perception, memory, and skill learning described. Extension of these causal relationships to physiological and neural levels is discussed. The general capability of a range of current consciousness models to support a modular hierarchy which could generate these causal relationships is reviewed, and the specific capabilities of two models with good general capabilities are compared in some detail. PMID- 15134762 TI - Locating volition. AB - In this paper, it is examined how neuroscience can help to understand the nature of volition by addressing the question whether volitions can be localized in the brain. Volitions, as acts of the will, are special mental events or activities by which an agent consciously and actively exercises her agency to voluntarily direct her thoughts and actions. If we can pinpoint when and where volitional events or activities occur in the brain and find out their neural underpinnings, this can substantively aid to demystify the concept of volition. After first discussing some methodological issues regarding whether it is possible to locate volition in the brain, various approaches by which neuroscientists and psychologists explore the neural correlates and substrates of volition are examined. Although different psychological conceptualizations of volition shape different perspectives toward understanding the functions of volition, the explorations of the neural basis of volition converge on certain common brain areas and structures. A unifying conception of volition that helps to make better sense of recent empirical findings is then suggested. PMID- 15134763 TI - From sensory processes to conscious perception. AB - In recent years, cognitive neuroscientists have began to explore the process of how sensory information gains access to awareness. To further probe this process, event-related functional magnetic resonance imaging (fMRI) was used while testing subjects with a paradigm known as the "attentional blink." In this paradigm, visually presented information sporadically fails to reach awareness. It was found that the magnitude and time course of activation within the anterior cingulate (BA 32), medial prefrontal cortex (BA 9), and frontopolar cortex (BA 10) predicted whether or not information was consciously perceived during the critical period for the attentional blink. These results are discussed in light of a neural framework for conscious processing. PMID- 15134764 TI - Implicit preferences: the role(s) of familiarity in the structural mere exposure effect. AB - In four experiments using an artificial grammar (AG) learning procedure, the authors examined the links between the "classic" mere exposure effect [heightened affect for previously encountered stimulus items (Bornstein, 1989; Zajonc, 1968)] and the "structural" mere exposure effect [greater hedonic appreciation for novel stimuli that conform to an implicitly acquired underlying rule system (Gordon & Holyoak, 1983)]. After learning, participants: (a) classified stimuli according to whether they conformed to the principles of the grammar and, (b) rated them in terms of how much they liked them. In some experiments unusual and unfamiliar symbols were used to instantiate the AG, in others highly familiar characters were used. In all cases participants showed standard AG learning. However, whether the two exposure effects emerged was dependent on symbol familiarity. Symbols with high a priori familiarity produced a structural mere exposure effect. Moderately familiar symbols produced only the classic, but not the structural, mere exposure effect. Highly unfamiliar symbols produced neither exposure effect. Results are discussed in the context of implicit learning theory and implications for a general theory of aesthetics are presented. PMID- 15134765 TI - No pause for a brief disruption: failures of visual awareness during ongoing events. AB - Past research has repeatedly documented the close relationship between visual attention and awareness. Most recently, research exploring change blindness, inattentional blindness, repetition blindness, and the attentional blink has converged on the conclusion that attention to one aspect of a scene or event may lead to a highly circumscribed awareness of only the specific information attended, while other information, even that which is spatially or temporally nearby can go completely unnoticed. In the present report, we extend these observations to the dynamic allocation of attention during a well-structured meaningful event. In two experiments, subjects viewed brief videos of simple events and were told to pay close attention to them. During the events, an unexpected disruption consisting of a brief low spatial frequency motion field occurred. Despite intensive questioning and opportunities for recognition, the majority of subjects reported no awareness of 200, 400, or 600 ms disruptions. In a second experiment, blank-screen disruptions were added, and these resulted in no increase in detection. We conclude that visual attention may result in far more transitory awareness of visual information than previously appreciated. PMID- 15134766 TI - Action, arousal, and subjective time. AB - Saccadic chronostasis refers to the subjective temporal lengthening of the first visual stimulus perceived after an eye movement. It has been quantified using a duration discrimination task. Most models of human duration discrimination hypothesise an internal clock. These models could explain chronostasis as a transient increase in internal clock speed due to arousal following a saccade, leading to temporal overestimation. Two experiments are described which addressed this hypothesis by parametrically varying the duration of the stimuli that are being judged. Changes in internal clock speed predict chronostasis effects proportional to stimulus duration. No evidence for proportionality was found. Two further experiments assessed the appropriateness of the control conditions employed. Results indicated that the chronostasis effect is constant across a wide range of stimulus durations and does not reflect the pattern of visual stimulation experienced during a saccade, suggesting that arousal is not critical. Instead, alternative processes, such as one affecting the onset of timing (i.e., the time of internal clock switch closure) are implicated. Further research is required to select between these alternatives. PMID- 15134767 TI - The influence of sex versus sex-related traits on long-term memory for gist and detail from an emotional story. AB - Recent findings demonstrate sex-related differences in the neurobiological mechanisms by which emotional arousal influences memory, and raise questions about the extent to which memory for emotional events may differ between males and females. Here we examine whether sex-related differences exist in the recall of central (gist) information and peripheral detail from an emotional story. Healthy subjects viewed a brief, narrated slide-show containing emotional elements in its middle section. One week later, they received an incidental multiple-choice recognition test for the story. Following the test, each subject completed the BEM Sex-Role Inventory, an assessment of sex-related masculine and feminine traits. The results reveal no differences in recall of either central or peripheral story information when considering the performance of actual men and women, but a significant difference when considering male and females as determined by their BEM test scores. "BEM" males (subjects with net male BEM scores) showed significantly enhanced recall of central emotional information. "BEM" females did not. Both groups showed significantly enhanced recall of peripheral emotional information, although this effect appeared larger in BEM females than in BEM males. The influences of "BEM" sex and type of information (central, peripheral) significantly interacted to influence emotional memory performance. These findings confirm the existence of sex-related influences in the recall of emotional information, and suggest that sex-related traits, rather than actual sex per se, may be a more sensitive indicator of these influences. PMID- 15134768 TI - Psychological and physiological characteristics of a proposed object referral/self-referral continuum of self-awareness. AB - This research extends and confirms recent brainwave findings that distinguished an individual's sense-of-self along an Object-referral/Self-referral Continuum of self-awareness. Subjects were interviewed and were given tests measuring inner/outer orientation, moral reasoning, anxiety, and personality. Scores on the psychological tests were factor analyzed. The first unrotated PCA component of the test scores yielded a "Consciousness Factor," analogous to the intelligence "g" factor, which accounted for over half of the variance among groups. Analysis of unstructured interviews of these subjects revealed fundamentally different descriptions of self-awareness. Individuals who described themselves in terms of concrete cognitive and behavioral processes (predominantly Object-referral mode) exhibited lower Consciousness Factor scores, lower frontal EEG coherence, lower alpha and higher gamma power during tasks, and less efficient cortical preparatory responses (contingent negative variation). In contrast, individuals who described themselves in terms of an abstract, independent sense-of-self underlying thought, feeling and action (predominantly Self-referral mode) exhibited higher Consciousness Factor scores, higher frontal coherence, higher alpha and lower gamma power during tasks, and more efficient cortical responses. These data suggest that definable states of brain activity and subjective experiences exist, in addition to waking, sleeping and dreaming, that may be operationally defined by psychological and physiological measures along a continuum of Object-referral/Self-referral Continuum of self-awareness. PMID- 15134769 TI - In sight but out of mind: do competing views test the limits of perception without awareness? AB - Over a century's worth of research suggests that "perception" without awareness is a genuine phenomenon. However, relatively little research has explored the question of whether all visually presented information activates representations in long term memory without awareness. Two experiments explored the use of a figure-ground display consisting of competing views in which one view dominates the other such that subjects are (initially) unaware of the non-dominant view. Neither experiment provided evidence that the non-dominant view (an embedded word) activated its representation in long term memory when the subject failed to report being aware of the embedded word in that priming was not seen on a subsequent stem completion task. In contrast, priming was seen when subjects reported being aware of the embedded word. It is suggested that two competing figure-ground relations are not concurrently computed unconsciously. PMID- 15134770 TI - Comments on commentaries: Kihlstrom, Bachmann, Reingold, and Snodgrass. PMID- 15134772 TI - Towards a cellular definition of CD8+ T-cell memory: the role of CD4+ T-cell help in CD8+ T-cell responses. AB - Whereas the definition of B-cell memory is based on well-known cellular properties and differentiation steps, the process of T-cell memory generation was, until recently, less well understood. A series of recent reports, however, have drastically modified our notion of CD8(+) memory T cells. They show that, in addition to division, the generation of efficient memory cells requires a previously unknown differentiation process. As a whole, the generation of CD8(+) memory T cells appears to mimic the generation of memory B cells. Both processes depend on the help of CD4(+) T cells, they are irreversible, they have the same mechanism, and they occur progressively during the late expansion phase of the primary immune response. PMID- 15134773 TI - The CD8 isoform CD8alphaalpha is not a functional homologue of the TCR co receptor CD8alphabeta. AB - Although structurally similar, CD8alphabeta and CD8alphaalpha have notably diverted with regard to function. Whereas CD8alphabeta functions as a T-cell receptor (TCR) co-receptor on MHC-class-I-restricted thymocytes and mature T cells, CD8alphaalpha is unable to support conventional positive selection, and can be expressed on T cells independent of the MHC restriction of their TCR. CD8alphaalpha induction is consistent with antigenic stimulation through the TCR, and recent developments have now shown that CD8alphaalpha induced on agonist triggered immature thymocytes, antigenic-stimulated conventional CD8alphabeta T cells and mucosal T cells mediates the specific modulation of TCR activation signals to facilitate their survival and differentiation into various specialized T-cell subsets. PMID- 15134774 TI - Apoptosis and loss of virus-specific CD8+ T-cell memory. AB - CD8(+) T-cell memory to viruses is stable in the absence but volatile in the presence of other infections. Apoptotic events that occur early in acute infections delete pre-existing memory T cells, leaving the host with reduced memory (except for cross-reactive responses) to previously encountered viruses. Apoptotic events also silence the acute immune response, leaving the host with a residual population of memory T cells. Persistent infections can induce apoptotic deletions of memory T cells that are specific to the persisting virus and to previously encountered pathogens. PMID- 15134775 TI - Regulation of IgA synthesis at mucosal surfaces. AB - Immunoglobulin A is the main element of the humoral immune response that has been selected through evolution, together with innate mucosal defences, to provide protection against microbial antigens at mucosal surfaces. IgA responses are initiated in organized inductive structures, such as Peyer's patches and nasal associated lymphoid tissues, as well as diffuse effector tissues, such as gut lamina propria and nasal mucosa. Hypermutated secretory IgAs play a critical role in regulating the composition of the intestinal microflora. Dysregulation of gut homeostasis in IgA-deficient gut causes a continuous activation of the immune cells and induces inflammatory processes leading to lymphoneogenesis. Recent advances in this field include new insights into the role of IgA in the maintenance of gut homeostasis and the proposal of an updated model for the induction of IgA responses in the gut. PMID- 15134777 TI - Ligand-independent tonic signaling in B-cell receptor function. AB - The random and inherently imprecise process of V(D)J recombination is the foundation for generation of the B-cell receptor (BCR). Signals must be generated to trigger selective processes that retain cells expressing a functional BCR, and these signals must be antigen-independent to insure an unbiased and diverse pool of newly formed B cells. Moreover, BCR expression, and presumably signaling, is essential for the continued survival of the B cell. Although BCR signaling is generally thought to depend upon ligand-induced aggregation, recent studies argue that some aspects of BCR signaling occur independently of antigen, and, furthermore, these non-induced or 'tonic' signals are linked to specific cellular processes operating at multiple stages of B-cell development. The potential co existence of tonic and induced signaling suggests a unique aspect of BCR complexes, or at least an aspect of receptors that has previously been under appreciated. PMID- 15134778 TI - Fine-tuning lymphocyte regulation: what's new with tyrosine kinases and phosphatases? AB - Although the basic mechanisms of lymphocyte signaling have been established, recent studies have provided new insights into how fine-tuning the regulation of tyrosine kinases and phosphatases contributes to the delicate balance required for appropriate lymphocyte activation. Recent studies include new work on the roles of the immune synapse in regulating T-cell receptor signaling, the discovery of new functions for the Src-family kinase Fyn and the Tec kinase Itk, particularly in regulation of the actin cytoskeleton, and new insights into positive and negative feedback mechanisms in antigen receptor signaling. PMID- 15134779 TI - Adaptors and linkers in T and B cells. AB - By mediating non-covalent protein-protein interactions, adaptors organize and assemble the multimolecular signalling complexes that coordinate intracellular programs leading to the activation and differentiation of lymphocytes. The co ordinated interaction between adaptor and effector molecules is required for the propagation and dynamic modification of externally applied signals. Recent advances have been made regarding our understanding of how adaptors regulate signalling within lymphocytes. An unexpected function has been revealed for the well-known adaptor protein LAT in pre-B-cell receptor signalling. In addition, the adaptors BCAP, Bcl10, CARMA1 and Malt1 seem to regulate the development of particular B-cell subsets. In contrast to Shc, c-Cbl and LAT, which are involved in early signalling events, BCAP, Bcl10, CARMA1 and Malt1 seem to act more distally, by controlling NF-kappaB activation. Additional transmembrane adaptors, such as NTAL/LAB and LIME, have been identified and partially characterized. Finally, an involvement of the cytosolic adaptors ADAP, SKAP-55 and Cbl-b in the regulation of lymphocyte adhesion and migration has been demonstrated. PMID- 15134780 TI - Phosphoinositide 3-kinase and its targets in B-cell and T-cell signaling. AB - Phosphoinositide 3-kinase (PI3K) activation is essential for lymphocyte proliferation driven by receptors for antigen, costimulatory ligands and cytokines. The lipid products of PI3K contribute to the assembly of membrane associated signaling complexes by promoting recruitment of selected proteins from the cytoplasm. Many proteins possess domains that are able to bind selectively to PI3K products. Different 'PI3K effector' proteins are coupled to distinct biological responses, depending on cell type and on the receptor that is engaged. In B cells and T cells, Tec-family tyrosine kinases and Akt serine/threonine kinases are emerging as crucial mediators of proliferation and survival signals downstream of PI3K. Of particular interest is recent evidence that PI3K signaling controls increases in lymphocyte size and metabolic activity that accompany cell cycle progression. PMID- 15134781 TI - Emerging paradigms of T-cell co-stimulation. AB - The analysis of recent data reveals that T-cell co-stimulation is a hierarchical process with elements of mutual interdependence between individual co stimulators. The expression and function of co-stimulatory molecules is biased on various T-cell subsets and is dependent on the T-cell differentiation state. The classical paradigm of T-cell co-stimulation by professional antigen-presenting cells has to incorporate the newly recognized concept of T-cell co-stimulation in the interaction with peripheral tissues, such as endothelial or epithelial cells. The two signal paradigm of T-cell co-stimulation is being replaced by a multisignal integration concept of central and peripheral co-stimulation. PMID- 15134782 TI - The regulation of lymphocyte activation by inhibitory receptors. AB - The ability of activating immune recognition receptors on lymphocytes to regulate cellular activation and function can be profoundly altered by co-stimulation with inhibitory receptors. Inhibitory receptors, such as the MHC-recognizing inhibitory receptors expressed on NK cells and subpopulations of activated T cells, can fully block the generation of any cytotoxic function by targeting proximal signals. Inhibitory Fc receptors on B cells, macrophages and mast cells can influence their threshold for activation, but the induction of inhibitory Fc receptors also appears to play a major role in the attenuation of ongoing responses. The three identified groups of inhibitory B7-recognizing receptors (CTLA-4, PD-1 and BTLA) are only expressed on activated hematopoietic cells, thus exclusively regulating ongoing immune responses in lymphoid organs and the periphery. In each case, the integrated positive and negative regulatory events determine the nature of the functional response. PMID- 15134783 TI - Proteomics/genomics and signaling in lymphocytes. AB - Recent technological advances in genomics, proteomics and bioinformatics have offered new insights into the molecular mechanisms that underlie lymphocyte signaling and function, and the development of new tools in these areas has opened up new avenues for biological investigation. By adding a quantitative dimension to lymphocyte proteome profiling, molecular machines and spatiotemporal regulatory processes can now be analyzed using such discovery-driven approaches. Biologists employing genomic and proteomic tools are gathering data at increasing speed and their struggle to extract maximal biological information is helped by new software tools that enable the detailed comparison of multiple datasets. PMID- 15134784 TI - New views of the immunological synapse: variations in assembly and function. AB - The interaction of T cells with antigen-presenting cells results in the formation of a contact face, termed the immunological synapse. The prototypical dynamics of this process are well established and involve cessation of crawling, a highly fluid 'immature' synapse phase during which signaling is initiated, and ultimately the formation of a 'mature' synapse characterized by centralized and peripheral supramolecular activating complexes. Ongoing research is directed towards defining how these supramolecular assemblies are formed and, more importantly, to what end. With regard to the former, progress has been made in defining the order in which various molecules are recruited to signaling centers in prototypical settings. With regard to the latter, however, the issue now appears more complex, as both developmental changes in T cells and variations in the environment appear to modulate features of mature synapse development. Although many details of the immunological synapse have been established, emerging evidence suggests a great variability in the ultimate form of these contacts and their effects on T-cell functions. PMID- 15134785 TI - Lipid raft domains and protein networks in T-cell receptor signal transduction. AB - Activation of the T-cell antigen receptor (TCR) is a key event in triggering the physiological responses of T lymphocytes to antigen. The earliest TCR-evoked signalling steps, such as tyrosine phosphorylations, ras activation and induction of Ca(2+) fluxes, are initiated in the T-cell plasma membrane. It has been implicated that cholesterol- and sphingolipid-rich membrane domains, termed lipid rafts, form platforms for the regulation and transduction of TCR signals at the plasma membrane; however, recent experiments have now differentiated distinct roles for lipid-raft-mediated and protein-mediated interactions in the formation of TCR signalling membrane domains. PMID- 15134786 TI - Notch signaling in lymphocyte development and function. AB - Over the past few years, the crucial role of Notch signaling in multiple stages of T-cell development has become apparent. Recent studies have helped to define more precisely the functions and components of the Notch signaling pathway in T cell development, including during the T versus B fate decision and in early CD4( )CD8(-)double-negative thymocytes. In addition, new evidence points to a requirement for Notch2 in the development of marginal zone B cells. Finally, recent studies have provided our first glimpse into the complex and paradoxical roles of Notch signaling in the activation and differentiation of mature T cells. PMID- 15134787 TI - Protein kinase C family functions in B-cell activation. AB - Members of the protein kinase C (PKC) family play important but distinct roles in B-cell activation, as demonstrated by emerging genetic and biochemical studies. PKCbeta is indispensable for B-cell antigen receptor (BCR)-induced NF-kappaB activation and B-cell survival. Recent evidence indicates that PKCbeta might regulate inhibitor of kappaB kinase (IKK) and NF-kappaB activation through interaction with the CARMA1/Bcl10/MALT signaling complex in BCR microdomains. By contrast, the novel PKC isoform PKCdelta is specifically required to maintain the tolerance of self-reactive B cells. PMID- 15134788 TI - T-cell-receptor- and B-cell-receptor-mediated activation of NF-kappaB in lymphocytes. AB - B and T cells sense antigens through specific receptors, which, when activated, induce signalling cascades leading to the activation of a series of transcription factor families, such as NF-kappaB. These transcription factors control differentiation, cytokine production and proliferation, and they protect against apoptosis. Much progress has been made during the past two years in the understanding of the molecular events leading to NF-kappaB activation, but, although most of the molecules in this signalling cascade have now been identified, the detailed molecular events remain obscure; in particular regarding the molecules that specifically connect the T-cell receptor (TCR)- and B-cell receptor (BCR)-proximal adaptors and kinases to the central core of the NF-kappaB cascade, the IkappaB kinase complex. As these events are likely to be specific for both extremities of the signalling cascade (the TCR or BCR on one end, and NF kappaB target genes on the other) they will ultimately represent the best targets to specifically manipulate this response in lymphocytes. PMID- 15134789 TI - B-cell antigen receptor signaling requirements for targeting antigen to the MHC class II presentation pathway. AB - The ability of B lymphocytes to capture, process and present antigens to T cells is requisite for normal humoral immune responses and contributes to the pathogenesis of both B- and T-cell-mediated autoimmune diseases. B lymphocytes preferentially capture polyvalent antigens, which are capable of eliciting a coordinated series of cellular responses that ensure that even low-affinity antigens are productively captured. Polyvalency not only accelerates transit through the endocytic pathway but also induces a reorganization of the antigen processing compartment, activates degradative pathways and determines how antigenic peptides are presented to T cells. Similar changes are observed in maturing dendritic cells, indicating that some cellular responses to foreign antigens are conserved. PMID- 15134790 TI - Suicidality in a clinical and community sample of adults with mental retardation. AB - This paper will argue that suicidal ideation and suicidal gestures are evident in adults with mental retardation, including individuals not receiving mental health services currently, and that psychosocial correlates of suicidality are similar to those noted in the general population. Findings are based on structured interviews with 98 adults with mental retardation, with corroborative information from caregivers and clinical charts. One in three individuals reported that they think "life is not worth living" sometimes or a lot. Eleven percent of individuals reported previous suicide attempt(s). Twenty-three percent of informants were unaware of the current suicidal ideation that their family member/client was reporting. Individuals reporting suicidal ideation endorsed more loneliness, stress, anxiety and depression, along with less social support than other individuals, consistent with reports of suicidal individuals in the general population. Adults with mental retardation who report thinking that life is not worth living should be a target group for future suicide prevention efforts. More research is needed to better understand the risk factors and protective factors for suicidality in this population. PMID- 15134791 TI - Finding meaning in parenting a child with Asperger syndrome: correlates of sense making and benefit finding. AB - This study explored the nature of two construals of meaning, benefit finding and sense making, in parents of a child with Asperger syndrome, and examined relations between both meaning constructs and the Double ABCX family stress model variables (initial stressor and pile-up of demands, appraisal, social support, coping strategies and adjustment) [H.I. McCubbin, J.M. Patterson, Social Stress and the Family: Advances and Developments in Family Stress Theory and Research, Haworth, New York, 1983, pp. 7-37]. A total of 59 parents completed questionnaires. Content analyses of parents' responses to questions inquiring about gains and sense making explanations revealed 8 benefit and 12 sense making themes. Results of correlations indicated that one or more of the meaning variables were related to each of the Double ABCX model predictors of parental adjustment. The meaning variables were positively related to adaptive coping processes: social support, self-efficacy, and problem-focused and emotional approach coping strategies. PMID- 15134792 TI - Health care service provision for country people with developmental disability: an Australian perspective. AB - Priorities in the delivery of health care to Australian country people with developmental disability were explored through focus groups and interviews of key stakeholder groups from four rural/regional towns. Data were analysed for themes, which converged on lack of access to primary health care providers who lacked knowledge about and had poor attitudes towards developmental disability, discriminatory practices, the burden on support people, and communication difficulties between GPs and people with developmental disability. Potential strategies to address the apparent dissatisfaction with primary health care and GP concerns about working with this group were also evident in the data. In particular, open communication between stakeholders as a mechanism for collaborative problem solving and a strategy to avoid burn out for both support people and GPs is recommended. Facilitation of such communication can best occur through undergraduate training and participation of disability workers in activities of organisations that support GPs. PMID- 15134793 TI - Effects of Snoezelen room, Activities of Daily Living skills training, and Vocational skills training on aggression and self-injury by adults with mental retardation and mental illness. AB - Multi-sensory stimulation provided in a Snoezelen room is being used increasingly for individuals with mental retardation and mental illness to facilitate relaxation, provide enjoyment, and inhibit behavioral challenges. We observed aggressive and self-injurious behavior in three groups of 15 individuals with severe or profound mental retardation and mental illness before, during, and after being in a Snoezelen room. All participants were receiving psychotropic medication for their mental illness and function-derived behavioral interventions for aggression, self-injury, or both. Using a repeated measures counterbalanced design, each group of participants was rotated through three experimental conditions: Activities of Daily Living (ADL) skills training, Snoezelen, and Vocational skills training. All other treatment and training activities specified in each individual's person-centered plan were continued during the 10-week observational period. Both aggression and self-injury were lowest when the individuals were in a Snoezelen room, followed by Vocational skills training and ADL skills training. The levels in the Snoezelen room were significantly lower than in both the other conditions for aggression but only in ADL skills training for self-injury. The difference in levels before and after Snoezelen were statistically significant with self-injury but not with aggression. The order of conditions showed no significant effect on either behavior. Snoezelen may provide an effective context for reducing the occurrence of self-injury and aggression. PMID- 15134794 TI - Two methods for teaching simple visual discriminations to learners with severe disabilities. AB - Simple discriminations are involved in many functional skills; additionally, they are components of conditional discriminations (identity and arbitrary matching-to sample), which are involved in a wide array of other important performances. Many individuals with severe disabilities have difficulty acquiring simple discriminations with standard training procedures, such as differential reinforcement. Errorless training methods may be more effective with this population. We used multiple-probe designs to compare two potentially errorless procedures for teaching simple discriminations among three pairs of photos of preferred items (S+) and colored rectangles (S-) to three youths with severe disabilities. In Experiment 1, baseline trials conducted with differential reinforcement yielded near-chance performances on all stimulus sets. A progressive delayed prompt training procedure was then implemented, with stimuli presented flat on the tabletop for one participant and at a 45 degrees angle to the tabletop for the other participants. After 120 teaching trials, accuracy remained near chance. Next, a stimulus control shaping procedure was implemented using an adapted Wisconsin General Test Apparatus (WGTA), with stimuli at a 45 degrees angle to the tabletop. Accuracy increased when this procedure was implemented with each stimulus pair in succession. In Experiment 2, for the participant whose stimuli were presented flat on the tabletop during the progressive delayed prompt training procedure, baseline trials were presented on the WGTA as at the end of Experiment 1, with differential reinforcement; accuracy remained high. On probe trials with stimuli placed flat on the tabletop, accuracy decreased to near-chance levels, indicating that the orientation of the stimulus array was a controlling variable. PMID- 15134795 TI - The discovery, isolation and identification of aldosterone: reflections on emerging regulation and function. AB - This paper has a focus on the early history of aldosterone. The Taits take us on a chronological trawl through the history in which they had a first hand role and made a major contribution-their bioassay was in many ways the key. The gifted Swiss chemists made a critical contribution to the scale and isolation of larger amounts. This was international collaboration at its best. Developing technologies were utilised as crucial cutting edge applications in the advancing front, technology transfer before the word was invented. Measurement of aldosterone and angiotensin were crucial advances to the understanding of the regulation of the hormone. In the period 1960-2003, some 30,000 papers mentioned aldosterone as a keyword, even so advances on a larger scale were slow. I have indicated some of my own work with the Howard Florey team using the adrenal autotransplant in the conscious sheep. Recently, the understanding of the role of induced proteins, the flow on from the RALES trial and the development of eplerenone has revitalised the aldosterone field. PMID- 15134796 TI - Glomerulosa cell--a unique sensor of extracellular K+ concentration. AB - The adrenal glomerulosa cells is the cell type most sensitive to extracellular K+ in the mammalian organism. Its sensitivity to physiological increases in K+ concentration ([K+]) is due to the expression of the two-pore domain K+ channels TASK that gives rise to K+ conductance in the range of resting membrane potential (approximately equal to -80mV) and to mechanisms that reduce the activation threshold of T-type voltage-activated Ca2+ channels. Potassium-induced cytoplasmic Ca2+ signal activates adenylyl cyclase; induces and activates StAR, the protein that carries cholesterol to the inner mitochondrial membrane and also enhances the expression of aldosterone synthase. The cytoplasmic Ca2+ signal is transferred into the mitochondrial matrix and enhances the reduction of mitochondrial pyridine nucleotides. PMID- 15134797 TI - The evolution of aldosterone antagonists. AB - Since the isolation and purification of aldosterone from adrenal extracts 50 years ago (Experientia 9 (1953) 33), scientists have learned a great deal about how and where aldosterone acts, the factors that control its release, what is its role in the pathophysiology of cardiovascular disease, how to make and study aldosterone antagonists, and for what medical purposes these agents are useful. In this paper, we will discuss the evolution of aldosterone antagonists from the relatively nonselective spironolactone (Aldactone), to the highly selective eplerenone (Inspra). Eplerenone represents a molecule with improved steroid receptor selectivity and pharmacokinetic properties in man compared to spironolactone. Recent clinical results have demonstrated that these improvements translate into tolerability and efficacy in patients with cardiovascular disease. PMID- 15134798 TI - Primary aldosteronism--careful investigation is essential and rewarding. AB - Once considered rare, primary aldosteronism (PAL) is now regarded as the commonest potentially curable and specifically treatable form of hypertension. At Greenslopes Hospital Hypertension Unit (GHHU), the decision in 1991 to screen all (and not just hypokalemic or resistant) hypertensives by aldosterone/renin ratio (ARR) testing led to a 10-fold increase in detection rate of PAL and four-fold increase in removal rate of aldosterone-producing adenomas (APAs). The GHHU/Princess Alexandra Hospital Hypertension Unit PAL series stands at 977 patients and 250 APAs removed with hypertension cured in 50-60% (remainder improved). Reliable detection requires that interfering medications are withdrawn (or their effects considered) before ARR measurement, and reliable methods (such as fludrocortisone suppression testing) to confirm PAL. Adrenal venous sampling is the only dependable way to differentiate APA from bilateral adrenal hyperplasia. Genetic testing has facilitated detection of glucocorticoid remediable, familial PAL. Identification of mutations causing the more common familial variety described by GHHU in 1991 should further aid in detection of PAL. PMID- 15134799 TI - Genetic factors associated with volume-sensitive hypertension. AB - Defining the genetic basis of essential hypertension is most informative when the blood pressure regulation is correlated with physiologic mechanisms, e.g., responses of the renin-angiotensin aldosterone system (RAAS) in hypertensive subjects. The aldosterone response to angiotensin II (Ang II) on a low salt diet is influenced by gender, plasma renin activity, and most significantly, familial resemblance, but only in males and in post-menopausal females. There is familial aggregation of low-renin hypertension, no association with candidate genes of the RAAS, but, a highly significant association with polymorphisms in the alpha adducin gene. Finally, angiotensinogen (AGT) genotype effects renal and adrenal responses to Ang II in patients with hypertension. These results strongly suggest that in contrast to population-based studies that use hypertension as the phenotype, classifying patients by the variability in physiologic, mechanistic traits enhances the probability of identifying the genetic factors influencing a rise in blood pressure. PMID- 15134800 TI - The 45-year story of the development of an anti-aldosterone more specific than spironolactone. AB - At the early stage of its development in 1957, the daily dose of spironolactone necessary to improve various pathological conditions was not precisely determined and dose-dependent sexual side effects limited its long-term use. Prescription of high daily doses and absence of selectivity for the mineralocorticoid receptor explain these limitations. The 9-11alpha epoxy group added to mexrenone by the Ciba-Geigy chemists in 1984 and improved chemical synthesis at Searle, permitted the original international clinical development of a selective antagonist for high blood pressure and congestive heart failure treatment. This review deals with the main methodological issues of a 20-year biological and clinical development of eplerenone, the second antimineralocorticoid drug. The investigation of a large range of daily doses (25-400mg) initially selected in normal volunteers by the 9alpha-fluorohydrocortisone test has led to the conclusion that 50-100mg q.i.d. doses of eplerenone offer a favorable benefit/risk ratio in various patient populations by neutralization of the aldosterone effects on blood pressure and target organ damage. The absence of sexual side-effects has confirmed the clinical relevance of the initial biological hypothesis on the need for more selectivity at the androgen and progestogen receptor sites. Widening the distance between efficacy and adverse effects of an anti-mineralocorticoid drug will facilitate the long-term maintenance of a moderately negative sodium balance and a slightly positive potassium balance, while minimizing the direct effects of salt and aldosterone on the heart, vessels, brain, and kidneys. Wide use in unselected patients and additional controlled clinical trials are necessary to confirm the benefits expected from animal and clinical research given that a 45-year interval also characterizes the story of the Na-Cl cotransporter (NCC) blocker, chlorthalidone, from its initial clinical use to the demonstration of its beneficial effects on cardiovascular morbidity and mortality. PMID- 15134801 TI - Effect of aldosterone blockade in patients with systolic left ventricular dysfunction: implications of the RALES and EPHESUS studies. AB - Aldosterone blockade has been shown to be effective in reducing total mortality as well as hospitalization for heart failure in patients with systolic left ventricular dysfunction (SLVD) due to chronic heart failure and in patients with SLVD post acute myocardial infarction. The evidence for the effectiveness of aldosterone blockade in chronic heart failure comes from the randomized aldactone evaluation study (RALES) while that for patients post infarction from the eplerenone post acute myocardial infarction efficacy and survival study (EPHESUS). These studies suggest that mineralocorticoid receptor activation remains important despite the use of an angiotensin converting enzyme inhibitor/angiotensin receptor blocking (ARB) agent and a beta blocker. Increasing evidence suggest that aldosterone blockade has important effects not only on the kidney but on ventricular remodeling, myocardial fibrosis, autonomic balance, fibrinolysis, oxidative stress, and activation of the NF-kappaB and AP-1 signaling pathways. The results of these studies in patients with SLVD has important implications not only for patients with chronic heart failure and post infarction but also for the therapy of patients with essential hypertension and other cardiovascular diseases. PMID- 15134802 TI - Glomerulosa function and aldosterone synthesis in the rat. AB - Since its discovery, it has been generally assumed that the primary function of the zona glomerulosa of the adrenal cortex is the secretion of aldosterone. Taking evidence from the rat, and recognising that there is probably considerable species variation, I argue here that the glomerulosa in fact has many functions, including aldosterone synthesis, but is probably only a relatively poor de novo source of steroid. In vitro, the CYP11B2 (aldosterone synthase) of the glomerulosa can and does utilise as substrates products arising from CYP11B1 (11beta-hydroxylase) activity in fasciculata cells. Whether it does in vivo is open to question, but corticosterone and 18-hydroxydeoxycorticosterone are both present in circulating rat plasma at suitable concentrations. Such a mechanism would explain several inconsistencies in the literature, including the anomalous distribution of steroidogenic enzymes in the glomerulosa, the stimulation of CYP11B1 products by aldosterone secretagogues such as potassium ions or angiotensin II, the partial dependence of aldosterone secretion in vivo on an intact pituitary, the sensitivity of aldosterone secretion to tissue disruption in vitro, and the "late pathway" regulation of aldosterone synthesis. PMID- 15134803 TI - The regulation of aldosterone synthase expression. AB - Aldosterone, the primary human mineralocorticoid, is a major regulator of intravascular volume and blood pressure. The capacity of the adrenal gland to produce aldosterone is controlled, in large part, by the regulated transcription of CYP11B2, the gene encoding aldosterone synthase. Aldosterone synthase is responsible for the conversion of 11-deoxycorticosterone to aldosterone and is expressed only within the zona glomerulosa of the adrenal cortex. The development of new systems for in vitro studies of expression has helped define molecular mechanisms that regulate this enzyme and thus the capacity of the adrenal gland to produce aldosterone. Both potassium and angiotensin II (ANG II) increase intracellular calcium levels, which regulate expression of CYP11B2 through transcription factors that interact with defined sites in the 5'-flanking region of the gene. PMID- 15134804 TI - Vascular synthesis of aldosterone: role in hypertension. AB - The author showed direct evidence that blood vessels are aldosteronogenic. The expression of CYP11B2 mRNA and synthesis of vascular aldosterone were decreased in rats treated with angiotensin converting enzyme inhibitor. Angiotensin II increased production of aldosterone in blood vessels. Vascular aldosterone and CYP11B2 mRNA levels of 2-week-old SHRSPs were significantly increased compared with that in WKY rats of the same age. High sodium intake develops and accelerates vascular injury and cardiac hypertrophy in SHRSP. Plasma aldosterone concentrations and plasma renin concentration were decreased by high salt intake in SHRSP. Aldosterone production, the expression of CYP11B2 mRNA and type I angiotensin II receptor (ATiR) mRNA in blood vessels were significantly increased by high salt intake. These results suggest that high salt intake increases aldosterone production and expression of the ATiR mRNA in the vascular tissue in SHRSP, which may contribute to the development of malignant hypertension in salt loaded SHRSP. PMID- 15134805 TI - Aldosterone synthase deficiency and related disorders. AB - Aldosterone's main actions are to regulate intravascular volume and serum electrolytes by controlling sodium absorbtion and potassium excretion in the distal nephron. Inherited defects in aldosterone biosynthesis thus cause hypovolemia, hyponatremia and hyperkalemia. Defective aldosterone biosynthesis may be caused by congenital adrenal hyperplasia due to 21-hydroxylase (CYP21) deficiency, in which case cortisol biosynthesis is also affected, or as an isolated defect termed aldosterone synthase (corticosterone methyloxidase, CYP11B2) deficiency. Many mutations have been documented in each of these genes; in general enzymatic activity must be reduced to <1% of normal for aldosterone biosynthesis to be impaired. An additional form of familial hyperreninemic hypoaldosteronism has been described that is not due to mutations in CYP11B2, but its etiology remains to be elucidated. PMID- 15134806 TI - Structural determinants of agonist-induced signaling and regulation of the angiotensin AT1 receptor. AB - Angiotensin II (Ang II) regulates aldosterone secretion by stimulating inositol phosphate production and Ca(2+) signaling in adrenal glomerulosa cells via the G(q)-coupled AT(1) receptor, which is rapidly internalized upon agonist binding. Ang II also binds to the heptahelical AT(2) receptor, which neither activates inositol phosphate signaling nor undergoes receptor internalization. The differential behaviors of the AT(1) and AT(2) receptors were analyzed in chimeric angiotensin receptors created by swapping the second (IL2), the third (IL3) intracellular loops and/or the cytoplasmic tail (CT) between these receptors. When transiently expressed in COS-7 cells, the chimeric receptors showed only minor alterations in their ligand binding properties. Measurements of the internalization kinetics and inositol phosphate responses of chimeric AT(1A) receptors indicated that the CT is required for normal receptor internalization, and IL2 is a determinant of G protein activation. In addition, the amino-terminal portion of IL3 is required for both receptor functions. However, only substitution of IL2 impaired Ang II-induced ERK activation, suggesting that alternative mechanisms are responsible for ERK activation in signaling-deficient mutant AT(1) receptors. Substitution of IL2, IL3, or CT of the AT(1A) receptor into the AT(2) receptor sequence did not endow the latter with the ability to internalize or to mediate inositol phosphate signaling responses. These data suggest that the lack of receptor internalization and inositol phosphate signal generation by the AT(2) receptor is a consequence of its different activation mechanism, rather than the inability of its cytoplasmic domains to couple to intracellular effectors. PMID- 15134807 TI - The use of scanning ion conductance microscopy to image A6 cells. AB - BACKGROUND: Continuous high spatial resolution observations of living A6 cells would greatly aid the elucidation of the relationship between structure and function and facilitate the study of major physiological processes such as the mechanism of action of aldosterone. Unfortunately, observing the micro-structural and functional changes in the membrane of living cells is still a formidable challenge for a microscopist. METHOD: Scanning ion conductance microscopy (SICM), which uses a glass nanopipette as a sensitive probe, has been shown to be suitable for imaging non-conducting surfaces bathed in electrolytes. A specialized version of this microscopy has been developed by our group and has been applied to image live cells at high-resolution for the first time. This method can also be used in conjunction with patch clamping to study both anatomy and function and identify ion channels in single cells. RESULTS: This new microscopy provides high-resolution images of living renal cells which are comparable with those obtained by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Continuous 24h observations under normal physiological conditions showed how A6 kidney epithelial cells changed their height, volume, and reshaped their borders. The changes in cell area correlated with the density of microvilli on the surface. Surface microvilli density ranged from 0.5 microm( 2) for extended cells to 2.5 microm(2) for shrunk cells. Patch clamping of individual cells enabled anatomy and function to be correlated. CONCLUSIONS: Scanning ion conductance microscopy provides unique information about living cells that helps to understand cellular function. It has the potential to become a powerful tool for research on living renal cells. PMID- 15134808 TI - Salt-inducible kinase (SIK) isoforms: their involvement in steroidogenesis and adipogenesis. AB - The cloning of salt-inducible kinase-1 (SIK1) that was specifically expressed in the adrenal glands of high-salt diet-fed rats led to subsequent cloning of adipose-specific SIK2 and rather ubiquitous SIK3. The three enzymes constitute a novel serine/threonine kinase subfamily, a member of AMP-activated protein kinase (PKA) family. Physiological roles of SIK1 and SIK2 have been investigated. The SIK1 transcript was expressed very early in the ACTH-stimulated Y1 cells, even before the expression of transcripts for CYP11A and StAR protein. Forced expression of SIK1 inhibited the ACTH-dependent expression of CYP11A- and StAR protein-genes. Cotransfection assays employing CRE-reporter gene showed that SIK1 could repress the PKA-dependent activation of CRE by acting on the bZIP domain of the CRE-binding protein (CREB), though the target site of SIK1-mediated phosphorylation has yet to be determined. ACTH/PKA-dependent nucleocytoplasmic shuttling of SIK1 took place in Y1 cells, implying that the intracellular movement of SIK1 might be a physiologically important determining factor for regulation of steroidogenic gene expression in the early phase of ACTH stimulation. The SIK2 gene was expressed in 3T3-L1 cells at a very early stage of adipogenesis. SIK2 could phosphorylate Ser-794 of human insulin-receptor substrate-1 (IRS-1) in vitro as well as in vivo. In addition, the SIK2 activity in db/db mice adipose tissues was significantly higher than that in wild-type adipose. These results strongly suggest that SIK2 may play important role(s) in modulating the insulin-signaling cascade of adipocytes, and thus, may be involved in the development of insulin resistance. Taken together, these results suggest that the SIK isoforms regulate hormonal signal transduction in both adrenal and adipose tissues. PMID- 15134809 TI - The control by angiotensin II of cholesterol supply for aldosterone biosynthesis. AB - In the adrenal glomerulosa cell, aldosterone is synthesized from cholesterol, which is supplied to the cell and stored under the form of cholesterol esters, then hydrolyzed to be transferred to the mitochondrial outer membrane and finally transported to the inner membrane where the P450 side-chain cleavage enzyme will convert it to pregnenolone. Angiotensin II (AngII), one of the major physiological regulators of mineralocorticoid synthesis, appears to affect most of the steps along this cascade and thus to exert a powerful control over the use of cholesterol for aldosterone production. PMID- 15134810 TI - Inactivating mutations of the mineralocorticoid receptor in Type I pseudohypoaldosteronism. AB - Type I pseudohypoaldosteronism (PHA1) is a rare form of mineralocorticoid resistance characterized by neonatal renal salt wasting and failure to thrive. Typical biochemical features include high levels of plasma aldosterone and renin, hyponatremia and hyperkalemia. Different mutations of the human mineralocorticoid receptor (hMR) gene have been identified in subjects affected by the autosomal dominant or sporadic form of the disease. Our laboratory has investigated a large number of subjects with familial and sporadic PHA1. Several different mutations have been detected, which are localized in different coding exons of the hMR gene. These mutations either create truncated proteins, either affect specific amino acids involved in receptor function. In this paper, we review hMR mutations described to date in PHA1 and their functional characterization. We discuss the absence of mutations in some kindreds and the role of precise phenotypic and biological examination of patients to allow for identification of other genes potentially involved in the disease. PMID- 15134811 TI - Expression and function of the human mineralocorticoid receptor: lessons from transgenic mouse models. AB - The human mineralocorticoid receptor (hMR), a ligand-dependent transcription factor (NR3C2) which belongs to the nuclear receptor superfamily, mediates most of the known effects of aldosterone. Beside its involvement in the regulation of sodium balance and the control of blood pressure, aldosterone-hMR tandem also exerts important regulatory functions on the cardiovascular and central nervous systems. To study the molecular mechanisms involved in the tissue-specific expression of hMR and explore its functional implication in pathophysiology, transgenic mouse models have been generated using both targeted oncogenesis and MR overexpression. We have previously demonstrated that the transcription of hMR is directed by two alternative promoters, P1 and P2, which correspond to the 5' flanking regions of the untranslated exons 1alpha and 1beta of the hMR gene, respectively. Utilization of P1 and P2 to drive expression of the SV40 large T antigen (TAg) in transgenic mice led us (i) to determine distinct tissue-specific patterns of promoter usage; (ii) to identify novel sites of MR expression including brown adipose tissue, thus providing a new functional link between aldosterone and energy homeostasis; (iii) to generate original immortalized cell lines derived from numerous aldosterone-sensitive tissues most notably distal nephron, brown fat, skin, liver, lung, brain, heart, blood vessels and inner ear. These differentiated cell lines represent suitable models to further explore cell specific mineralocorticoid responses and cross-talk with other signaling pathways. Generation of transgenic mice in which hMR expression was directed by P1 promoter demonstrated the importance of MR in the cardiac and renal function. Morphological and functional alterations of the renal tubule were observed with basal decreased sodium/potassium ratio exacerbated under sodium depletion. Hypokinetic dilated cardiomyopathies were associated with tachycardia, arrhythmia but normal arterial blood pressure emphasizing the direct role of MR on cardiomyocyte function. Taken together, transgenic animal models constitute valuable experimental systems to gain new insights into the widespread and pleiotropic in vivo functions of MR. PMID- 15134812 TI - A quick glance at rapid aldosterone action. AB - Not all of the actions of aldosterone are mediated by the classic genomic pathway involving transcription and translation. Non-genomic or non-classical rapid responses that do not require these steps have been known for some time, but have only attracted significant interest in the last decade. At the cellular level, second messengers and kinase cascades are commonly involved. Most of these non classical effects are insensitive to inhibitors of the classical cytosolic mineralocorticoid receptor. Non-genomic aldosterone action has been observed in clinical studies particularly in the cardiovascular system, and further research may improve the understanding of their participation in the pathogenesis of aldosterone related diseases and eventually enhance the options for therapy. PMID- 15134813 TI - Molecular basis for the apparent mineralocorticoid excess syndrome in the Oman population. AB - 11beta-Hydroxysteroid dehydrogenase type 2 (11beta-HSD2) plays a crucial role in converting hormonally active cortisol to inactive cortisone, thereby conferring specificity upon the mineralocorticoid receptor (MR). Mutations in the gene encoding 11beta-HSD2 (HSD11B2) account for an inherited form of hypertension, the syndrome of "Apparent Mineralocorticoid Excess" (AME) where cortisol induces hypertension and hypokalaemia. We report five different mutations in the HSD11B2 gene in four families from Oman with a total of 9 affected children suffering from AME. Sequence data demonstrate the previously described L114Delta6nt mutation in exon 2 and new mutations in exon 3 (A221V), exon 5 (V322ins9nt) and for the first time in exon 1 (R74G and P75Delta1nt) of the HSD11B2 gene. These additional mutations provide further insight into AME and the function of the 11beta-HSD2 enzyme. The prevalence of monogenic forms of hypertension such as AME remains uncertain. However, our data suggests AME may be a relevant cause of hypertension in certain ethnic groups, such as the Oman population. PMID- 15134814 TI - Towards selectively modulating mineralocorticoid receptor function: lessons from other systems. AB - Although there is clinical utility in blocking mineralocorticoid receptor (MR) action, the usefulness of available MR antagonists is limited because of cross reactivity with the androgen and progesterone receptors (spironolactone) or possibly by low affinity for MR (eplerenone). MR binds aldosterone and physiologic glucocorticoids, such as cortisol, which both can act as MR agonists in epithelial tissues. However, in preliminary studies aldosterone and cortisol appear to induce different conformations in non-epithelial tissues; in the cardiomyocyte, cortisol usually acts as an MR antagonist, whereas in vascular smooth muscle cortisol mimics aldosterone actions if it can access MR, just as it does in the kidney. Thus, there are needs for improved MR antagonists with higher selectivity and potency and, if possible, for compounds that lock MR into specific desirable conformations. Efforts are underway to modulate selectively the action of many nuclear receptors, and insights from one nuclear receptor may be applicable to others given the similarities in structure and function. We have used traditional approaches aided by X-ray crystallography to obtain several classes of selective ligands that modulate thyroid receptor (TR) action. We describe the properties of these selective TR modulators here, and discuss the possibility that similar approaches to ligand design may yield MR interacting compounds with improved specificity and, possibly, tissue specificity. PMID- 15134815 TI - Molecular mechanism of activation and nuclear translocation of the mineralocorticoid receptor upon binding of pregnanesteroids. AB - The mineralocorticoid receptor (MR) is primarily localized in the cytoplasm of the cell in the absence of ligand. The first step in the genomic-dependent mechanism of action of mineralocorticoids is the binding of steroid to the MR, which in turn triggers MR nuclear translocation. The regulation of hormone binding to MR is complex and involves a multifactorial mechanism, making it difficult to determine the optimal structure of a steroid for activating the MR and promoting its nuclear translocation. Here we review the structure-activity relationship for several pregnanesteroids that possess various functional groups, and suggest that a flat conformation of the ligand rather than the presence of particular chemical groups is a critical parameter for the final biological effect in vivo. We also discuss how the MR undergoes differential conformational changes according to the nature of the bound ligand, which in turn affects the dynein-dependent retrograde rate of movement for the steroid/receptor complex. PMID- 15134816 TI - Identification of steroid ligands able to inactivate the mineralocorticoid receptor harboring the S810L mutation responsible for a severe form of hypertension. AB - The ability of steroid ligands to inactivate the human mineralocorticoid receptor (MR(WT)) has been shown to be due to their inability to contact Asn770, a residue of the H3 helix involved in stabilizing contacts with the H11-H12 loop region. However, all steroid ligands that display antagonist properties when bound to MR(WT), have been shown to activate a mutant receptor (MR(L810)) associated with a severe form of hypertension. Biochemical studies revealed that S810L mutation induces a change in the receptor conformation and increases the steroid-receptor complexes stability. From a three-dimensional model of the MR ligand-binding domain, it is likely that the S810L mutation causes a steric hindrance between the side chains of Leu810 (H5) and Gln776 (H3) that provokes a bending of the H3 helix. As a consequence, the positioning of MR(WT) antagonists within the ligand binding cavity is modified in such a way that they can activate the mutant MR(L810). The results from biochemical studies also revealed that 5alpha-pregnan 20-one, 4,9-androstadiene-3,17-dione and RU486, unable to bind MR(WT), acted as potent MR(L810) antagonists. PMID- 15134817 TI - Regulation of epithelial ion transport by aldosterone through changes in gene expression. AB - The year 2003 marks the 50th year since the unfolding of the chemical structures of both aldosterone and DNA. Since the recognition in the early 1960's that aldosterone and its cousin cortisol act through DNA binding proteins that alter gene transcription, research on these corticosteroid hormones and their receptors has attracted fervent attention, both for their importance in endocrine physiology, and as model systems for understanding gene regulation. Recently, aldosterone has emerged as arguably the single most important physiological regulator of extracellular fluid volume and blood pressure in mammals, and has been implicated in a variety of disease states in humans. Moreover, its principal receptor, the mineralocorticoid receptor is increasingly recognized as an important therapeutic target for the treatment of hypertension and congestive heart failure, as well as an important model system for understanding aspects of gene regulation. This increased insight into the functional and pathophysiologic importance of aldosterone has been accompanied by increased insight into its cellular and molecular mechanisms of action. Aldosterone acts in a variety of epithelial and non-epithelial tissues to influence extracellular fluid volume, blood pressure, salt appetite, and can under the appropriate conditions cause cardiac fibrosis. This review will address the current view of aldosterone's molecular mechanism of action in epithelia focusing primarily on the classical MR and on a particular MR target gene, SGK1. PMID- 15134818 TI - Regulation of sodium transport in mammalian collecting duct cells by aldosterone induced kinase, SGK1: structure/function studies. AB - Serum- and glucocorticoid-induced kinases (SGK) are members of the serine threonine kinase family. SGK1, the isoform identified first, is rapidly induced by aldosterone. In this study, we determined that the two recently described isoforms, SGK2 and SGK3 are also expressed in renal cortical collecting duct (CCD) cells; however, their expression is not induced by aldosterone or glucocorticoids. SGK1 increases the activity of the epithelial sodium channel (ENaC) in oocytes but its cellular targets in native mineralocorticoid target cells and its mechanism of action are still unknown. We studied the role of SGK1 in corticosteroid-regulated Na transport in M-1 mouse CCD cell lines that stably over-express or down-regulate SGK1. Basal rates of transepithelial Na transport were significantly lower in CCD cells in which SGK1 expression or activity was down-regulated than in SGK1 overexpressing cells. Importantly, corticosteroid treatment failed to stimulate Na transport in cells with down-regulated SGK1 while it significantly increased Na transport in parent and SGK1 overexpressing M 1 cells. To determine if C-terminal PDZ interactions are important for SGK's effect on ENaC activity or trafficking, we examined the effects of mutant SGK1 in which the conserved PDZ binding domain has been eliminated. However, such mutations did not decrease its stimulatory effect on ENaC current in Xenopus oocytes. Fluorescence confocal microscopy revealed that the intracellular localization of full-length and PDZ binding mutated SGK1 was identical: they both localize to intracellular vesicular structures. On the other hand, N-terminally truncated (delta 60)-SGK1 did not increase ENaC activity. We conclude that SGK1 is a critical component in corticosteroid-regulated Na transport in mammalian CCD cells. Our data also indicate that the N-terminal of SGK1 is necessary for its stimulatory effect on Na transport while elimination of the C-terminal PDZ binding domain did not change its function. PMID- 15134819 TI - Mineralocorticoid receptor binding, structure and function. AB - The isolation of aldosterone 50 years ago was a critical first step in elucidating the mechanism by which corticosteroids regulate electrolyte homeostasis. The broad principles of this mechanism involving an intracellular receptor acting on specific genes to induce the expression/repression of aldosterone-induced proteins (AIP) were established 30 years ago. The cloning of the mineralocorticoid receptor (MR) has enabled studies of the subcellular mechanisms of aldosterone action, including the molecular dissection of structure function relationships in the receptor. We have exploited the close structural and functional similarity of the MR with the glucocorticoid receptor to identify the regions in the MR that confer ligand-binding specificity. The critical region is located, not as might be expected in the ligand-binding pocket but rather on the surface of the molecule. These studies have been extended to an analysis of the interactions between the N-terminal and ligand-binding domains of the MR. In the last decade, AIP have been identified; the regulation of the genes encoding these AIP are discussed. PMID- 15134820 TI - Aldosterone-synthase overexpression in heart: a tool to explore aldosterone's effects. AB - Clinical observations indicate that elevated aldosterone impairs cardiovascular function. The mechanisms, however, are not totally understood although total and cardiovascular mortality are decreased by aldosterone antagonists. Experimentally, increased plasma aldosterone induces pericoronary inflammation and cardiac fibrosis. Our laboratory has discovered that aldosterone is synthesized in the rat heart, and has demonstrated that this cardiac aldosterone is involved in post-infarction cardiac remodeling. In man, activated cardiac aldosterone production has been described in patients with heart failure. In transgenic mice that overexpress aldosterone-synthase in the heart, we observe a normal cardiac function but a major coronary dysfunction, more pronounced in males. These observations converge to a potential physiological and pathological relevance of this system. Beneficial effects of anti-aldosterone treatment in heart failure may thus be secondary in part to blockade of cardiac aldosterone action. PMID- 15134821 TI - The myosin binding protein is a novel mineralocorticoid receptor binding partner. AB - The mineralocorticoid receptor (MR) plays a role in congestive heart failure; however, the molecular mechanism(s) remains undefined. We hypothesized that interaction of the MR with a cardiac protein modulates the transcriptional activation function of the MR within the heart. We used the yeast two-hybrid technique to screen a human heart library and found an aldosterone-dependent interaction between the hMR and the cardiac myosin binding protein (cMBP-c). The EC(50) of the hMR-MBP-c interaction was approximately 80nM, and the cMBP-c did not interact with the glucocorticoid receptor (GR). The GST pull-down technique was used to confirm an interaction between the MR and the cMBP-c as well as the lack of interaction with the GR. Spironolactone partially blocked this interaction, further suggesting MR specificity. We also determined the cMBP-c binding site lies within the C-terminus of the MR. We propose that interaction of the MR with cMBP-c may play a role in cardiac remodeling. PMID- 15134822 TI - Aldosterone target organ protection by eplerenone. AB - The classical mineralocorticoid effect of aldosterone on unidirectional transepithelial sodium transport in the kidney was long thought to be the predominant effect of this hormone. However, there is convincing evidence for additional extrarenal actions of aldosterone that are mediated via activation of mineralocorticoid receptors (MRs) in the heart, vasculature and brain. It is now postulated that many of the detrimental effects of aldosterone are mediated through MR activation in these nonclassical target organs. The selective aldosterone blocker, eplerenone (Inspra), is under development for human therapeutic use for treatment of hypertension and heart failure post-myocardial infarction (MI). Clinical and preclinical studies have linked elevated aldosterone to hypertension, left ventricular and vascular remodeling, cardiac, renal, and cerebral vascular inflammation and injury, and increased risk of mortality in heart failure patients. Multiple studies in experimental models of hypertension and heart failure demonstrate that selective blockade of aldosterone by eplerenone effectively preserves cardiac function, attenuates maladaptive left ventricular remodeling and tissue and vascular injury in part by reducing vascular inflammation in aldosterone target organs. PMID- 15134823 TI - Aldosterone-induced vasculopathy. AB - Aldosterone antagonists reduce mortality in chronic heart failure (CHF). An obvious question to ask is how do they do this. The prevailing hypothesis is that most of the adverse effects of aldosterone stem from its ability to produce a vasculopathy. This vasculopathy is characterised by a reduction in vascular nitric oxide and may be produced by aldosterone's ability to generate superoxide radicals which degrade endogenous NO. The consequences of this "aldosterone induced vasculopathy" are that it produces tissue ischaemia/infarction and injury, which then repairs itself by producing fibrosis. "Aldosterone-induced vasculopathy" may be the main mechanism why aldosterone promotes widespread tissue injury and ultimately cardiac death. PMID- 15134824 TI - The impact of polymorphisms in the gene encoding aldosterone synthase (CYP11B2) on steroid synthesis and blood pressure regulation. AB - The terminal stages in the synthesis of aldosterone and cortisol are catalysed by the enzymes aldosterone synthase and 11beta-hydroxylase respectively. We have previously reported that polymorphic variation in the 5' promoter region ( 344C/T) of the gene encoding aldosterone synthase (CYP11B2) is associated with increased aldosterone metabolite excretion and with hypertension associated with a raised aldosterone to renin ratio (ARR). Additionally, basal and ACTH stimulated plasma levels of 11-deoxycortisol, the precursor of cortisol, are higher in subjects carrying the T-allelic variant. We have now identified in a family study (573 individuals from 105 extended families ascertained through a hypertensive proband) that excretion of the main metabolite of this steroid (tetrahydro-11-deoxycortisol, THS) is heritable (19.4%) and that the T-allele of CYP11B2 is more strongly associated with higher THS levels than the C-allele. Raised plasma and urinary levels of 11-deoxycortisol suggest that there is relative inefficiency of 11beta-hydroxylation in the zona fasciculata; the P450 enzyme responsible for this step is encoded by the gene CYP11B1, which is highly homologous with and adjacent to CYP11B2. The association of genetic variation in the promoter of CYP11B2 which, in the adrenal cortex, is only expressed in zona glomerulosa, and zona fasciculata 11beta-hydroxylation function is paradoxical. There may be linkage dys-equilibrium between this polymorphism and a quantitative trait locus (QTL) in CYP11B1. Chronic alteration of 11beta-hydroxylase activity may increase ACTH drive to the adrenal cortex, altering the regulation of aldosterone synthesis. This may explain, at least partly, the association between CYP11B2 polymorphisms and hypertension. PMID- 15134825 TI - Development of test systems for the discovery of selective human aldosterone synthase (CYP11B2) and 11beta-hydroxylase (CYP11B1) inhibitors. Discovery of a new lead compound for the therapy of congestive heart failure, myocardial fibrosis and hypertension. AB - Two key players in adrenal steroid hormone biosynthesis are the human mitochondrial cytochrome P450 enzymes CYP11B1 and CYP11B2 that catalyze the final steps in the biosynthesis of cortisol and aldosterone, respectively. Overproduction of both hormones contributes to a number of severe diseases, as illustrated by the association of elevated aldosterone levels with hypertension and higher mortality in congestive heart failure, and by Cushing's syndrome as the clinical correlate of chronic hypercortisolism. Thus, CYP11B1 and CYP11B2 comprise new targets for drug treatment and selective inhibitors of both enzymes are of high pharmacological interest. To facilitate the search for such compounds, we have established novel test procedures using recombinant fission yeast strains that stably express these enzymes. The aim of this study was to compare the inhibition profiles displayed by these enzymes in established mammalian cell culture test systems to those obtained with the new fission yeast assays, and to evaluate the usefulness of the Schizosaccharomyces pombe strains as screening systems for the identification of novel lead compounds. Using these test systems, we were able to identify a new and very selective CYP11B2 inhibitor (SIAS-1) that displayed no detectable interference with CYP11B1 activity. PMID- 15134826 TI - Drospirenone, a progestogen with antimineralocorticoid properties: a short review. AB - Progesterone (P) has high affinity to the mineralocorticoid receptor (MCR), and it is an MCR antagonist. Almost all synthetic progestogens are devoid of this antimineralocorticoid (anti-MC) effect. They are unable to antagonize the salt retaining effect of estrogens. This could be one cause of weight gain and an increase in blood pressure with the use of combined oral contraceptives (OC) and, in some susceptible women, with postmenopausal estrogen/(progestogen) treatment. The purpose of this presentation is to review results of clinical studies with drospirenone (DRSP), a new progestogen developed by Schering A.G., with anti-MCR activity. DRSP is a derivative of 17-alpha-spirolactone. In rats, rabbits and in man, it is a PR-agonist and an MCR- and androgen-R antagonist with no effect on the glucocorticoid-R and the estrogen-R. In normally menstruating women, 2-3mg DRSP per day, taken from day 5 to 25 of the cycle, inhibit ovulation, lead to a mild natriuresis, and a slight compensatory activation of the renin-aldosterone system. Compared with an OC containing 30 microg ethinylestradiol (EE) and 150 microg levonorgestrel, a combination of 3mg DRSP with 30 microg EE given over 6 months led to a slight decrease in body weight and blood pressure. The reduction in mean body weight by a combination of DRSP with EE compared with a conventional OC could be confirmed in a study over 26 months in 900 young women. The OC containing DRSP has favorable effects in patients suffering from the premenstrual syndrome (PMS), and, partly due to the antiandrogenic effect of DRSP, in those with acne vulgaris. For postmenopausal women, a combination of DRSP with estradiol has been developed with the expectation that the slight blood pressure lowering and weight reducing effects will minimize cardiovascular morbidity in patients needing hormone treatment because of hot flushes and other climacteric symptoms. CONCLUSIONS: DRSP, by its anti-MCR effect and its potential to slightly decrease body weight and blood pressure, shares many pharmacodynamic properties with progesterone, and it is a candidate for reducing cardiovascular morbidity in women using OCs or postmenopausal hormone treatment. PMID- 15134828 TI - Proceedings of the International Symposium on Aldosterone. London, United Kingdom, April 28-30, 2003. PMID- 15134827 TI - Aldosterone, mineralocorticoid receptors and vascular inflammation. AB - For the past 50 years, the physiological action of aldosterone was considered to be on epithelial tissues to maintain fluid and electrolyte homeostasis. Recently, a nonepithelial, pathophysiologic, proinflammatory role for aldosterone has been inferred from studies on mineralocorticoid/salt administration, with or without mineralocorticoid receptor (MR) blockade, in experimental animals, and from clinical studies such as RALES and EPHESUS. More recently still, it has become clear that the pathophysiologic trigger for the vascular inflammatory response observed is not necessarily aldosterone per se, but inappropriate activation of vascular wall MR. MR can be inappropriately activated by aldosterone in the context of an inappropriate salt status, or by glucocorticoids in the context of tissue damage. The studies supporting this latter conclusion, and the novel mechanisms proposed to support this concept, are details in the text to follow. PMID- 15134829 TI - Use of high affinity insulin analogues to assess the functional relationships between insulin receptor trafficking, mitogenic signaling and mRNA expression in rat liver. AB - We have investigated the functional relationships between insulin receptor (IR) trafficking, mitogenic signaling and mRNA expression in rat liver and primary hepatocytes. The low-K(d) insulin analogues [His(A8),His(B4), Glu(B10),His(B27)] human insulin (-HI) (the H2-analogue), [Asp(B10)]HI and [Glu(A13),Glu(B10)]HI, were studied in liver parenchymal cells and compared with wild-type HI and epidermal growth factor (EGF), a mitogenic inducer. The extent and duration of IR endocytosis were markedly increased in response to the H2-analogue and [Asp(B10)]HI compared to wild-type HI, but similar to HI after [Glu(A13),Glu(B10)]HI administration. Importantly, the insulin analogues induced a higher and more prolonged tyrosine phosphorylation of the IR-beta subunit in endosomes compared to authentic HI. A low cell-free endosome-lysosome transfer of the internalized IR was only observed in response to HI and H2-analogue injection. Concomitant with the low endosome-lysosome transfer of the intact IR beta subunit, 47 and 50 kDa fragments of the IR-beta subunit accumulated in lysosomal fractions. Neither HI nor the insulin analogues promoted the endosomal recruitment and tyrosine phosphorylation of Shc, whereas EGF accessed the Shc signaling pathway. Moreover, EGF induced a fast and prolonged activation of Raf-1 and MAP-kinase pathways whereas HI and insulin analogues displayed a moderate and transient effect. Finally, treatment of primary rat hepatocytes with HI and the protease-resistant H2-analogue did not affect the total level and relative expression of isotype A and B of IR mRNA regardless of time of exposure. These results suggest a lack of relationship between IR trafficking, endosomal tyrosine phosphorylation and mitogenic signaling in rat liver in vivo. PMID- 15134830 TI - Three-dimensional structure of an unusual Kunitz (STI) type trypsin inhibitor from Copaifera langsdorffii. AB - The crystallographic structure of a novel trypsin inhibitor (CTI) from Copaifera langsdorffii is reported. The structure was solved by MIRAS procedure and refined to a crystallographic residual of 17.3% (R(free) = 20.3%) at 1.8 A resolution. Two isomorphous derivatives were obtained by quick cryo-soaking approach. CTI is the first structure of a member of Kunitz (STI) family formed by two noncovalently bound polypeptide chains and only one disulfide bridge. A standard Kunitz-type inhibitor has a single polypeptide chain and two disulfide bridges. Structural features granting CTI high inhibitory activity are discussed. PMID- 15134831 TI - Integrin alpha v beta 3 binds a unique non-RGD site near the C-terminus of human tropoelastin. AB - Tropoelastin is the soluble precursor of the essential resilient connective tissue protein elastin. We examined the binding of integrin alpha(v)beta(3) to tropoelastin. In quantitative colorimetric solid-phase assays, purified alpha(v)beta(3) demonstrated saturable, divalent cation-dependent, single-site binding behavior on tropoelastin with a dissociation constant of 3.8 +/- 0.9 nM in the presence of 1 mM Mn(2+) which increased to 23 +/- 5 nM in the presence of 1 mM Ca(2+). Association with alpha(v)beta(3) was localized to the C-terminal 16 residues of tropoelastin, encompassing the region encoded by exon 36. This region comprises a unique disulfide loop in tropoelastin that is not essential for the interaction. This is the first identification of a specific, single binding site on tropoelastin and the first observation of direct binding of an integrin to a tropoelastin domain. PMID- 15134832 TI - The adaptor disabled-2 binds to the third psi xNPxY sequence on the cytoplasmic tail of megalin. AB - The cytoplasmic tail (CT) of megalin possesses several functional motifs likely to participate in protein-protein interactions within the proximal tubular epithelial cell (PTEC) of the kidney. One such interaction is with the phosphotyrosine interaction domain (PID) of the adaptor protein disabled-2 (Dab2), a mitogen-responsive phosphoprotein, which interacts via its PID with Psi xNPxY (where Psi represents a hydrophobic residue) motifs on its binding partners. Megalin CT has three such motifs; it has been established that there is no interaction of Dab2 with the first (from N to C) (Biochem. J. 3 (2000) 613). Here, we analyse in real-time the binding of recombinant megalin CT, and of synthetic peptide sequences encompassing the second and third Psi xNPxY motifs, to Dab2PID in real-time using surface plasmon resonance (SPR). We report a binding affinity of DabPID for megalin CT of K(D) = 2.6 x 10(-7) +/- 5.3 x 10( 8). Direct binding and competition studies indicate that this interaction is with the third Psi xNPxY motif. The dissociation of Dab2 from the third Psi xNPxY peptide was significantly slower than that from the second (k(off) (mean +/- S.E.M.) (per s) = 0.002 +/- 0.002 vs. 0.007 +/- 0.002, P < 0.05). Synthetic peptide sequences encompassing the third Psi xNPxY but not the second inhibited Dab2PID binding both to intact megalin CT and to the third Psi xNPxY motif. Tyrosine phosphorylation of either motif did not exert a major effect upon competition efficacy. We further demonstrate for the first time the presence of Dab2 expression in primary human PTEC. PMID- 15134833 TI - On the evolution of erythrocyte programmed cell death: apoptosis of Rana esculenta nucleated red blood cells involves cysteine proteinase activation and mitochondrion permeabilization. AB - Batracian Rana esculenta erythrocytes cell death induced by either calcium influx, or staurosporine, involves typical apoptotic phenotype. Our data reveal: (i) a drastic modification of the cell morphology with loss of the ellipsoidal form as assessed by phase contrast microscopy and scanning electron microscopy; (ii) an exposure of the phosphatidylserine residues in the outer leaflet of the cell membrane; (iii) a caspase-3-like activity; (iv) a mitochondrial membrane potential (Delta Psi m) loss; and (v) a chromatin condensation and fragmentation. Erythrocyte chromatin condensation and fragmentation are prevented by caspase and calpain peptide inhibitors. These inhibitors also prevent Delta Psi m loss supporting the idea that mitochondria is a central sensor for Rana erythrocytes cell death. Our observations highlight the conservation of the programmed cell death machinery in erythrocytes across kingdom. PMID- 15134834 TI - Isolation and characterization of hyaluronidase a "spreading factor" from Indian cobra (Naja naja) venom. AB - Hyaluronidase, ubiquitous enzyme in snake venoms, known originally as "spreading factor", has not been well studied. The present study describes the purification and characterization of hyaluronidase from Indian cobra (Naja naja) venom and provides systematic evaluation of the spreading property of the enzyme. Hyaluronidase (NNH1) has been purified through gel permeation and ion exchange chromatography. The molecular mass was found to be 70.406 kDa by MALDI-TOF mass spectrometry and with the (p)i pI of 9.2. The amino acid sequence of the N terminus was found to be NEQSTHGAYV. The enzyme shows absolute specificity for hyaluronan and belongs to the group of neutral active enzymes. Tetrasaccharides are the final product of hyaluronan digestion. The enzyme cleaves beta 1,4 glycosidic linkage and belongs to a group of endo-beta-N-acetyl hexosaminidases. Hyaluronidase indirectly potentiates the myotoxicity of VRV-PL-VIII, a phospholipolytic myotoxin, and also the hemorrhagic potency of a hemorrhagic complex-I. Localization of hyaluronan in human skin section and selective degradation by venom hyaluronidase (NNH1) corroborate the plausible in vivo degradation of hyaluronan in the extracellular matrix (ECM) resulting in easy dissemination of VRV-PL-VIII myotoxin and hemorrhagic complex-I. PMID- 15134835 TI - Purification of a Class B1 platelet aggregation inhibitor phospholipase A2 from Indian cobra (Naja Naja) venom. AB - A platelet aggregation inhibitor phospholipase A(2) (NND-IV-PLA(2)) was isolated from Naja naja (Eastern India) venom by a combination of cation and anion exchange chromatography. NND-IV-PLA(2) is the most catalytically active enzyme isolated from the Indian cobra venom. The acidic PLA(2) profile of Eastern regional Indian cobra venom is distinctly different from that of the western regional venom. However the acidic PLA(2)s from both the regions follow the pattern of increasing catalytic activity with increase in acidic nature of the PLA(2) isoform. NND-IV-PLA(2) is a Class B1 platelet aggregation inhibitor and inhibits platelet aggregation induced by ADP, collagen and epinephrine. Modification of active site histidine abolishes both catalytic activity and platelet aggregation inhibition activities while aristolochic acid, a phospholipase A(2) inhibitor has only partial effect on the two activities. PMID- 15134836 TI - Analysis of Bothrops jararacussu venomous gland transcriptome focusing on structural and functional aspects: I--gene expression profile of highly expressed phospholipases A2. AB - Snake venom glands are a rich source of bioactive molecules such as peptides, proteins and enzymes that show important pharmacological activity leading to in local and systemic effects as pain, edema, bleeding and muscle necrosis. Most studies on pharmacologically active peptides and proteins from snake venoms have been concerned with isolation and structure elucidation through methods of classical biochemistry. As an attempt to examine the transcripts expressed in the venom gland of Bothrops jararacussu and to unveil the toxicological and pharmacological potential of its products at the molecular level, we generated 549 expressed sequence tags (ESTs) from a directional cDNA library. Sequences obtained from single-pass sequencing of randomly selected cDNA clones could be identified by similarities searches on existing databases, resulting in 197 sequences with significant similarity to phospholipase A(2) (PLA(2)), of which 83.2% were Lys49-PLA(2) homologs (BOJU-I), 0.1% were basic Asp49-PLA(2)s (BOJU II) and 0.6% were acidic Asp49-PLA(2)s (BOJU-III). Adjoining this very abundant class of proteins we found 88 transcripts codifying for putative sequences of metalloproteases, which after clustering and assembling resulted in three full length sequences: BOJUMET-I, BOJUMET-II and BOJUMET-III; as well as 25 transcripts related to C-type lectin like protein including a full-length cDNA of a putative galactose binding C-type lectin and a cluster of eight serine proteases transcripts including a full-length cDNA of a putative serine protease. Among the full-length sequenced clones we identified a nerve growth factor (Bj NGF) with 92% identity with a human NGF (NGHUBM) and an acidic phospholipase A(2) (BthA-I-PLA(2)) displaying 85-93% identity with other snake venom toxins. Genetic distance among PLA(2)s from Bothrops species were evaluated by phylogenetic analysis. Furthermore, analysis of full-length putative Lys49-PLA(2) through molecular modeling showed conserved structural domains, allowing the characterization of those proteins as group II PLA(2)s. The constructed cDNA library provides molecular clones harboring sequences that can be used to probe directly the genetic material from gland venom of other snake species. Expression of complete cDNAs or their modified derivatives will be useful for elucidation of the structure-function relationships of these toxins and peptides of biotechnological interest. PMID- 15134837 TI - Proteoglycans in human laryngeal cartilage. Identification of proteoglycan types in successive cartilage extracts with particular reference to aggregating proteoglycans. AB - The content, composition and structure of proteoglycans (PGs) in adult human laryngeal cartilage (HLC) were investigated. PGs were extracted from the tissue by using two different extraction protocols. In the first protocol, PGs were extracted under dissociative conditions, 4 M guanidine HCl (GdnHCl), and in the second protocol, sequentially, with phosphate buffered saline (PBS) and solutions of increasing GdnHCl concentration (0.5, 1, 2 and 4 M). Chemical and immunological analyses of dissociate extracts (first protocol) revealed the presence of four, at least, different types of PGs. Aggrecan was the major PG, versican, decorin and biglycan being in small amounts. Galactosaminoglycan containing PGs (GalAGPGs) represented the vast majority of total PGs present in extracts of HLC. Differential digestion with chondroitinase ABC and AC II showed that the GalAGPGs from HLC contained a significant proportion of dermatan sulphate (DS). In addition, disaccharide analysis showed that 6-sulphated disaccharides predominated in chondroitin sulphate (CS) chains. The sequential extraction (second protocol) indicated that PBS extract contained very little amount of PGs. The 0.5, 1 and 2 M GdnHCl extracts contained 6.3%, 24.5% and 15.2% of total extracted PGs, respectively. Four molar GdnHCl extracted the larger proportion, about 53%, of total PGs. This extract contained almost only proteoglycan aggregate components i.e., G1 bearing aggrecan, hyaluronan and link protein. The characterization of the aggrecan showed that it constituted a polydisperse population of monomers with an average molecular mass of 720 kDa. The glycosaminoglycans (GAGs) present were chondroitin sulphate with a M(r) of 15 kDa, and keratan sulphate (KS) with a M(r) of 10 kDa, in proportions 84% and 16%, respectively. PMID- 15134838 TI - A quantitative in silico analysis calculates the angiotensin I converting enzyme (ACE) inhibitory activity in pea and whey protein digests. AB - Angiotensin I converting enzyme (ACE) inhibitory peptides can induce antihypertensive effects after oral administration. By means of an ACE inhibitory peptide database, containing about 500 reported sequences and their IC(50) values, the different proteins in pea and whey were quantitatively evaluated as precursors for ACE inhibitory peptides. This analysis was combined with experimental data from the evolution in ACE inhibitory activity and protein degradation during in vitro gastrointestinal digestion. Pea proteins produced similar in silico scores and were degraded early in the in vitro digestion. High ACE inhibitory activity was observed after the simulated stomach phase and augmented slightly in the simulated small intestine phase. The major whey protein beta-lactoglobulin obtained the highest in silico scores, which corresponded with the fact that degradation of this protein in vitro only occurred from the simulated small intestine phase on and resulted in a 10-fold increase in the ACE inhibitory activity. Whey protein obtained total in silico scores of about 124 ml/mg, compared to 46 ml/mg for pea protein, indicating that whey protein would be a richer source of ACE inhibitory peptides than pea protein. Although beta lactoglobulin is only partially digested, a higher ACE inhibitory activity was indeed found in the whey (IC(50) = 0.048 mg/ml) compared to the pea digest (IC(50) = 0.076 mg/ml). In silico gastrointestinal digestion of the highest scoring proteins in pea and whey, vicilin and albumin PA2, and beta lactoglobulin, respectively, directly released a number of potent ACE inhibitory peptides. Several other ACE inhibitory sequences resisted in silico digestion by gastrointestinal proteases. Briefly, the quantitative in silico analysis will facilitate the study of precursor proteins on a large scale and the specific release of bioactive peptides. PMID- 15134839 TI - Development of orbitofrontal function: current themes and future directions. AB - In recent years, an exciting thrust in the developmental research literature has been the focus on "executive" functions (EF). However, the emphasis has been on the more purely cognitive aspects of EF operative in abstract reasoning and problem solving-aspects associated mainly with dorsolateral frontal regions. Although the literature on adult neuropsychology has seen an emerging and growing interest in the study of processes more related to orbitofrontal cortex (OFC), including emotion and personality, research on the development of EF has lagged behind. This special issue of Brain and Cognition is intended to redress this imbalance by bringing together researchers who are studying the nature and development of orbitofrontal function from a wide variety of perspectives. PMID- 15134840 TI - The functions of the orbitofrontal cortex. AB - The orbitofrontal cortex contains the secondary taste cortex, in which the reward value of taste is represented. It also contains the secondary and tertiary olfactory cortical areas, in which information about the identity and also about the reward value of odours is represented. The orbitofrontal cortex also receives information about the sight of objects from the temporal lobe cortical visual areas, and neurons in it learn and reverse the visual stimulus to which they respond when the association of the visual stimulus with a primary reinforcing stimulus (such as taste) is reversed. This is an example of stimulus reinforcement association learning, and is a type of stimulus-stimulus association learning. More generally, the stimulus might be a visual or olfactory stimulus, and the primary (unlearned) positive or negative reinforcer a taste or touch. A somatosensory input is revealed by neurons that respond to the texture of food in the mouth, including a population that responds to the mouth feel of fat. In complementary neuroimaging studies in humans, it is being found that areas of the orbitofrontal cortex are activated by pleasant touch, by painful touch, by taste, by smell, and by more abstract reinforcers such as winning or losing money. Damage to the orbitofrontal cortex can impair the learning and reversal of stimulus-reinforcement associations, and thus the correction of behavioural responses when there are no longer appropriate because previous reinforcement contingencies change. The information which reaches the orbitofrontal cortex for these functions includes information about faces, and damage to the orbitofrontal cortex can impair face (and voice) expression identification. This evidence thus shows that the orbitofrontal cortex is involved in decoding and representing some primary reinforcers such as taste and touch; in learning and reversing associations of visual and other stimuli to these primary reinforcers; and in controlling and correcting reward-related and punishment-related behavior, and thus in emotion. The approach described here is aimed at providing a fundamental understanding of how the orbitofrontal cortex actually functions, and thus in how it is involved in motivational behavior such as feeding and drinking, in emotional behavior, and in social behavior. PMID- 15134841 TI - The role of emotion in decision-making: evidence from neurological patients with orbitofrontal damage. AB - Most theories of choice assume that decisions derive from an assessment of the future outcomes of various options and alternatives through some type of cost benefit analyses. The influence of emotions on decision-making is largely ignored. The studies of decision-making in neurological patients who can no longer process emotional information normally suggest that people make judgments not only by evaluating the consequences and their probability of occurring, but also and even sometimes primarily at a gut or emotional level. Lesions of the ventromedial (which includes the orbitofrontal) sector of the prefrontal cortex interfere with the normal processing of "somatic" or emotional signals, while sparing most basic cognitive functions. Such damage leads to impairments in the decision-making process, which seriously compromise the quality of decisions in daily life. The aim of this paper is to review evidence in support of "The Somatic Marker Hypothesis," which provides a systems-level neuroanatomical and cognitive framework for decision-making and suggests that the process of decision making depends in many important ways on neural substrates that regulate homeostasis, emotion, and feeling. The implications of this theoretical framework for the normal and abnormal development of the orbitofrontal cortex are also discussed. PMID- 15134842 TI - The neuropsychology of ventral prefrontal cortex: decision-making and reversal learning. AB - Converging evidence from human lesion, animal lesion, and human functional neuroimaging studies implicates overlapping neural circuitry in ventral prefrontal cortex in decision-making and reversal learning. The ascending 5-HT and dopamine neurotransmitter systems have a modulatory role in both processes. There is accumulating evidence that measures of decision-making and reversal learning may be useful as functional markers of ventral prefrontal cortex integrity in psychiatric and neurological disorders. Whilst existing measures of decision-making may have superior sensitivity, reversal learning may offer superior selectivity, particularly within prefrontal cortex. Effective decision making on existing measures requires the ability to adapt behaviour on the basis of changes in emotional significance, and this may underlie the shared neural circuitry with reversal learning. PMID- 15134843 TI - Autobiographical memory and the self in time: brain lesion effects, functional neuroanatomy, and lifespan development. AB - Autobiographical remembering reflects an advanced state of consciousness that mediates awareness of the self as continuous across time. In naturalistic autobiographical memory, self-aware recollection of temporally and spatially specific episodes and generic factual information (both public and personal) operate in tandem. Evidence from both laboratory and real-life studies, however, suggests that these two processes can be dissociated. This paper reviews aging, lesion, and functional neuroimaging research on the anatomical substrates of autobiographical memory processes using a new measure, the Autobiographical Interview, and prospective collection of autobiographical material. Results indicate that autobiographical recollection is mediated by a distributed fronto temporo-parietal system, with the anteromedial prefrontal cortex positioned to integrate sensory information with self-specific information. The emergence of autobiographical recollection at around age four coincides with the timing of prefrontal regressive cortical and progressive white matter changes that may support the development of this high-level capacity. PMID- 15134844 TI - The frontal lobes and theory of mind: developmental concepts from adult focal lesion research. AB - The primary objective in this paper is to present a framework to understand the structure of consciousness. We argue that consciousness has been difficult to define because there are different kinds of consciousness, hierarchically organized, which need to be differentiated. Our framework is based on evidence from adult focal lesion research. The different types of consciousness are associated with distinct brain regions, with the higher levels of consciousness related to self-awareness and theory of mind (both facets of consciousness), with an emphasis on the role of the frontal lobes. The secondary objective is to use this structure to suggest hypotheses about the potential effect of frontal dysfunction at various developmental stages, and including both congenital and acquired brain injury. PMID- 15134845 TI - Developmental outcomes after early prefrontal cortex damage. AB - The neuropsychological bases of cognitive, social, and moral development are minimally understood, with a seemingly wide chasm between developmental theories and brain maturation models. As one approach to bridging ideas in these areas, we review 10 cases of early prefrontal cortex damage from the clinical literature, highlighting overall clinical profiles and real life developmental outcomes. Based on these cases, there is preliminary evidence to support distinctive developmental differences after: (1) dorsolateral, (2) mesial, and (3) orbital polar prefrontal lesions, for more profound impairments after bilateral damage, and possibly for recovery differences after very early vs. later childhood lesion onset. Further case and group studies are needed to confirm reliable effects of specific lesion locations, the influence of age of lesion onset, and related experiential and treatment variables in determining adult outcomes. Rather than a single underlying deficit associated with early prefrontal cortex damage, we interpret the findings to suggest that it is the altered integration and interplay of cognitive, emotional, self-regulatory, and executive/metacognitive deficits that contribute to diverse developmental frontal lobe syndromes. The findings support the fundamental importance of prefrontal cortex maturation in protracted cognitive, social-emotional, and moral development. PMID- 15134846 TI - Plasticity and functions of the orbital frontal cortex. AB - We compare the effects of psychoactive drugs such as morphine and amphetamine on the synaptic organization of neurons in the orbital frontal (OFC) and medial frontal (mPFC) regions in the rat. Both regions are altered chronically by exposure to intermittent doses of either drug but the effects are area-dependent. For example, whereas morphine produces increased spine density in OFC but decreased spine density in mPFC. The differential response of the OFC and mPFC to drugs is paralleled by an areal-dependent effect of gonadal hormones on these regions as well: males have greater dendritic arborization in the mPFC whereas females have a greater arborization in the OFC. We also compared the effects of neonatal injury to the OFC and mPFC on cognitive, motor, and social behaviors as well as on the anatomical organization of the remaining brain. Again, there were differential effects of the treatments to the OFC and mPFC. Neonatal OFC lesions allowed virtually complete functional recovery of cognitive and motor behaviors, which was correlated with mild abnormalities in cerebral development compared to the more severe deficits and morphological sequelae following mPFC lesions at the same ages. One exception was the effect of OFC on social behavior, which was severe regardless of whether the injury was in infancy or adulthood. It is proposed that both drug-induced and developmental abnormalities in the integrity of OFC neurons may lead to deficits in social behavior or other behavioral pathologies, possibly including depression. PMID- 15134847 TI - Charting the maturation of the frontal lobe: an electrophysiological strategy. AB - Tracking the functional development of specific regions of the prefrontal cortex in children using event-related potentials (ERPs) is challenging for both technical and conceptual reasons. In this paper we outline our strategy for studying frontal lobe development and present preliminary results from children aged 7-17 years and young adults using ERPs functionally associated with anterior cingulate and prefrontal cortex, especially the orbitofrontal, ventral, and medial portions. Our analysis of contingent negative variation, error-related negativity, and novelty P300 data show that the ERPs associated with these regions are still maturing into late adolescence, and that their amplitude has significant correlations with behavioral capacities. PMID- 15134848 TI - Sex differences in early childhood, adolescence, and adulthood on cognitive tasks that rely on orbital prefrontal cortex. AB - Through the use of several tests of cognition we have documented sex differences in young children, adolescents, and adults on tasks that rely on the integrity of the orbital prefrontal cortex. In children under three years of age, males performed with significantly fewer errors than did females on tests of object reversals. No significant sex differences were found in older children, despite the use of a more challenging object reversal task. Sex differences were also found in adolescents and adults on the Iowa Gambling Task. On this decision making task, in contrast to males, females appear to be responding to different elements of the task. Discussion of the implications for these findings is presented. PMID- 15134849 TI - Development of "hot" executive function: the children's gambling task. AB - Development of affective decision-making was studied in 48 children at two ages (3 and 4 years) using a simplified version of the Iowa Gambling Task (). On each of 50 trials, children chose from 1 of 2 decks of cards that, when turned, displayed happy and sad faces, corresponding to rewards (candies) won and lost, respectively. Cards in 1 deck offered more rewards per trial, but were disadvantageous across trials due to occasional large losses; cards in the other deck offered fewer rewards per trial, but were advantageous overall. On later trials, 4-year-olds made more advantageous choices than 3-year-olds, and 4-year olds made more advantageous choices than would be expected by chance, whereas 3 year-olds made more disadvantageous choices than would be expected by chance. These findings, which were especially pronounced for girls, indicate that affective decision-making develops rapidly during the preschool period, possibly reflecting the growth of neural systems involving orbitofrontal cortex. PMID- 15134850 TI - Complex decision-making in early childhood. AB - Decision-making over time is an important aspect of adaptive social functioning. The main goal of this study was to investigate the development of this ability in young children. A simplified version of the Iowa Gambling Task was given to 69 children at 3 ages (3, 4, and 6 years). Children were also given an awareness test to assess their knowledge of the task. Significant age differences were found for awareness of the task while significant sex effects were found for performance on the task. Females chose significantly more from the advantageous decks than would be expected by chance in the second block. Males demonstrated no significant difference in choice of decks. Further analysis indicated female superiority in the task was not due to greater knowledge of the game. One interpretation of these results is that there are two systems affecting decision making over time. PMID- 15134851 TI - Resistance to extinction: a measure of orbitofrontal function suitable for children? AB - Extinction of operantly conditioned responses, which provides a measure of the ability to adapt to changes in the reinforcement value of stimuli, has been linked to orbitofrontal cortex (OFC) in human and non-human animals. This article examines the feasibility of using extinction as a measure of the development of OFC function in preschool-age children. If extinction serves as a measure of OFC function, resistance to extinction should decrease during this age range. In Part 1, we review the literature on extinction as related to OFC, and summarize what is known regarding age-related changes in extinction in young children. In Part 2, we report results of a study assessing extinction in children between 3 and 6 years of age. Results revealed age-related increases in responding during extinction, with girls (particularly, 6-year-old girls) exhibiting stronger resistance to extinction. Some relations were found with temperament and normative compulsive-like behavior. The findings of this study question the utility of extinction of operant responding as a measure of OFC function in young children. PMID- 15134853 TI - The roles of orbital frontal cortex in the modulation of antisocial behavior. AB - This article considers potential roles of orbital frontal cortex in the modulation of antisocial behavior. Two forms of aggression are distinguished: reactive aggression elicited in response to frustration/threat and goal directed, instrumental aggression. It is suggested that orbital frontal cortex is directly involved in the modulation of reactive aggression. It is argued that orbital frontal cortex does not "inhibit" reactive aggression but rather may both increase or decrease its probability as a function of social cues present in the environment. Early dysfunction in this function of orbital frontal cortex may be linked to the development of Borderline Personality Disorder. Instrumental aggression is linked to a fundamental failure in moral socialization. However, the available data suggest that the amygdala, but not orbital frontal cortex, is required for functions such as aversive conditioning and passive avoidance learning that are necessary for moral socialization. Psychopathic individuals who present with significant instrumental aggression, are impaired in aversive conditioning and passive avoidance learning and show evidence of amygdala dysfunction. Orbital frontal cortex and the amygdala are involved in response reversal where instrumental responses must be reversed following contingency change. Impairments in response reversal are also seen in psychopathic individuals. However, it remains unclear whether impairment in response reversal per se is associated with antisocial behavior. PMID- 15134854 TI - Understanding orbitofrontal contributions to theory-of-mind reasoning: implications for autism. AB - Autism is a lifelong developmental disorder that is associated with severe difficulties with "theory-of-mind"--the understanding that others' behaviors are motivated by internal mental states. Here, we raise the possibility that research examining the neural bases of theory-of-mind reasoning has the potential to inform researchers about the elusive functional neural impairments associated with autism. Evidence from our lab and others' suggests that theory-of-mind reasoning may be fractionated into at least two functionally and anatomically distinct neural circuits. Specifically, the ability to decode others' mental states from observable cues (such as facial expressions) may rely on contributions from the orbitofrontal/medial temporal circuit within the right hemisphere. In contrast, the ability to reason about others' mental states may rely left medial frontal regions. We conclude by reviewing evidence suggesting that the developmental roots of autism might lie in abnormal functioning of the orbitofrontal/medial temporal circuit which may, in turn, underlie the abnormal development of social-cognitive skills among individuals with autism. PMID- 15134852 TI - Neurocognitive elements of antisocial behavior: Relevance of an orbitofrontal cortex account. AB - This paper reviews the role of orbitofrontal cortex (OFC) lesions in antisocial behaviors and the adequacy of a strict OFC account of antisocial disorders where there is no evidence of lesion. Neurocognitive accounts of antisocial behaviors are extended beyond the OFC. Several methodological shortcomings specific to this neuroscience approach to antisocial behavior are identified. A developmental approach is advocated to chart the developmental sequences of impaired brain development and of the various comorbid states typically seen in antisocial disorders. PMID- 15134855 TI - The role of the orbitofrontal cortex in normally developing compulsive-like behaviors and obsessive-compulsive disorder. AB - Mounting evidence concerning obsessive-compulsive disorders points to abnormal functioning of the orbitofrontal cortices. First, patients with obsessive compulsive disorder (OCD) perform poorly on tasks that rely on response suppression/motor inhibition functions mediated by the orbitofrontal cortex relative to both normal and clinical controls. Second, patients with OCD exhibit functional hyperactivity in lateral orbitofrontal and related structures corresponding with symptom severity. In this article, we compare these neurocognitive correlates of OCD with the executive and neural underpinnings of "compulsive-like" behaviors that are common in normal childhood. We discuss the phenomenology and natural history of normative compulsive-like behaviors as well as the behavioral, emotional, and cognitive continuities between typical and pathological obsessive-compulsive behaviors. We then examine associations between children's executive performance deficits and their observed compulsive-like characteristics. We relate these patterns to executive deficits shown by adults with OCD. Finally, we speculate on the developmental neurobiology of children's compulsive-like behaviors, with particular attention to orbitofrontal functions including behavioral and emotional regulation, and we suggest similarities and differences with the neurobiology of OCD. In making these comparisons, we hope to open a dialogue between researchers who study underlying brain pathologies associated with OCD and those who explore the neurocognitive bases of normal development. PMID- 15134856 TI - Twenty-five years of peptides. PMID- 15134857 TI - Corticotropin releasing hormone receptors: two decades later. AB - Hypothalamic corticotropin releasing hormone (CRH) regulates pituitary ACTH secretion and mediates behavioral and autonomic responses to stress, through interaction with type 1 plasma membrane receptors (CRHR1) located in pituitary corticotrophs and the brain. Although the CHRI are essential for ACTH responses to stress, their number in the pituitary gland does not correlate with corticotroph responsiveness, suggesting that activation of a small number of receptors is sufficient for maximum ACTH production. CRH binding and hybridization studies in adrenalectomized, glucocorticoid-treated or stressed rats revealed divergent changes in CRH receptors and CRH1 mRNA in the pituitary, with a reduction in receptor binding but normal or elevated expression of CHR1 mRNA levels. Western blot analysis of CRHR1 protein in pituitary membranes from adrenalectomized rats showed unchanged receptor mRNA levels and increased CRHR1 protein, despite binding down-regulation, suggesting that decreased binding is due to homologous desensitization, rather than reduced receptor synthesis. In contrast, decreased CRH binding following glucocorticoid administration is associated with a reduction in CRHR1 protein, suggesting inhibition of CRH1 mRNA translation. The regulation of CRHR1 translation may involve binding of cytosolic proteins, and a minicistron in the 5'-UTR of the CRHR1 mRNA. It is likely that post-transcriptional regulatory mechanisms that permit rapid changes in CRH receptor activity are important for adaptation of corticotroph responsiveness to continuous changes in physiological demands. PMID- 15134858 TI - The many lives of leptin. AB - Leptin is a 16,000-Da protein which is secreted by fat but acts within the brain to regulate adiposity. Our Peptides Classic addressed the mystery of how such a large molecule could negotiate the blood-brain barrier (BBB), a structure which normally excludes proteins from the brain. We found that leptin was transported across the BBB by a saturable transport system. This finding was important to understanding how satiety-related peptides and proteins worked, but it was also important to the concept that the BBB is a regulatory interface important in brain-body communication. Obesity in humans and many animals is associated with a leptin resistant state rather than a leptin deficiency. Subsequent work has shown that a defect in the BBB transport of leptin is key in producing and reinforcing this state of resistance. Leptin is pluripotent and the concept of it being primarily an adipostat is being discarded for more encompassing views. Consideration of the BBB data would favor the view that ancestral levels of leptin were much lower than those currently considered normal and are consistent with leptin acting as a metabolic switch, informing the brain when fat reserves are adequate to direct energy expenditures towards activities other than seeking calories. PMID- 15134859 TI - A 25 year adventure in the field of tachykinins. AB - Several aspects of our 25 year adventure in the field of tachykinins will be successively described. They concern: substance P (SP) synthesis and release in the basal ganglia, the identification and pharmacological characterization of central tachykinin NK(1), NK(2) and NK(3) binding sites and their topographical distribution, the description of some new biological tests for corresponding receptors, the identification of tachykinin NK(1) receptor subtypes or conformers sensitive to all endogenous tachykinins (substance P, neurokinin A (NKA), neurokinin B (NKB), neuropeptide gamma (NP gamma) and neuropeptide K (NPK)) and finally, the functional involvement of these receptors and their subtypes in tachykinin-induced regulations of dopamine and acetylcholine release in the striatum. PMID- 15134860 TI - Neuropeptide Y, ubiquitous and elusive. AB - This paper reviews aspects of NPY research that were emerging in 1985, shortly after the isolation and characterization of the peptide. NPY had become known for its widespread distribution especially in the central and peripheral nervous systems, but also in the gastro-intestinal and respiratory tracts and in fibers innervating smooth muscle around blood vessels. Consistent with its distribution, it was determined that NPY is a potent vasoconstrictor, affects neuroendocrine systems and is involved in appetite regulation--areas of research still relevant today. Through advances in technology knowledge about NPY's role in these and newly discovered physiological functions has deepened considerably. Successful cloning of a series of NPY receptors has opened up new and complex research vistas. Lately, the creation of mice genetically modified for NPY as well as for several receptor subtypes has brought many puzzling observations--followed by questions yet to be answered. PMID- 15134861 TI - Receptor autoradiography as mean to explore the possible functional relevance of neuropeptides: focus on new agonists and antagonists to study natriuretic peptides, neuropeptide Y and calcitonin gene-related peptides. AB - Over the past 20 years, receptor autoradiography has proven most useful to provide clues as to the role of various families of peptides expressed in the brain. Early on, we used this method to investigate the possible roles of various brain peptides. Natriuretic peptide (NP), neuropeptide Y (NPY) and calcitonin (CT) peptide families are widely distributed in the peripheral and central nervous system and induced multiple biological effects by activating plasma membrane receptor proteins. The NP family includes atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP) and C-type natriuretic peptide (CNP). The NPY family is composed of at least three peptides NPY, peptide YY (PYY) and the pancreatic polypeptides (PPs). The CT family includes CT, calcitonin gene-related peptide (CGRP), amylin (AMY), adrenomedullin (AM) and two newly isolated peptides, intermedin and calcitonin receptor-stimulating peptide (CRSP). Using quantitative receptor autoradiography as well as selective agonists and antagonists for each peptide family, in vivo and in vitro assays revealed complex pharmacological responses and radioligand binding profile. The existence of heterogeneous populations of NP, NPY and CT/CGRP receptors has been confirmed by cloning. Three NP receptors have been cloned. One is a single-transmembrane clearance receptor (NPR-C) while the other two known as CG-A (or NPR-A) and CG-B (or NPR-B) are coupled to guanylate cyclase. Five NPY receptors have been cloned designated as Y(1), Y(2), Y(4), Y(5) and y(6). All NPY receptors belong to the seven-transmembrane G-protein coupled receptors family (GPCRs; subfamily type I). CGRP, AMY and AM receptors are complexes which include a GPCR (the CT receptor or CTR and calcitonin receptor-like receptor or CRLR) and a single-transmembrane domain protein known as receptor-activity-modifying-proteins (RAMPs) as well as an intracellular protein named receptor-component-protein (RCP). We review here tools that are currently available in order to target each NP, NPY and CT/CGRP receptor subtype and establish their respective pathophysiological relevance. PMID- 15134862 TI - Neurotransmitters co-existing with VIP or PACAP. AB - It is now recognized that a neuron can produce, store and release more than one transmitter substance, and a number of examples of co-existing transmitters, particularly a neuropeptide together with a classical transmitter, have been reported. The present paper deals with transmitter substances, peptides or classical transmitters, co-existing with the two structurally related peptides VIP and PACAP and the possible functional implications of this co-existence. PMID- 15134863 TI - Dynamic neuronal-glial interactions: an overview 20 years later. AB - After commenting on some perceived reasons why our review may have been relatively frequently cited, a brief overview is presented that first summarizes what we knew 25 years ago about the dynamic neuronal-astroglial interactions that occur in response to changes in the physiological state of the animal. The brain system in which these dynamic interactions were studied was the magnocellular hypothalamo-neurohypophysial system (mHNS) of the rat. The mHNS developed as and continues to be the model system yielding the most coherent picture of dynamic morphological changes and insights into their functional consequences. Many other brain areas, however, have more recently come under scrutiny in the search for glial-neuronal dynamisms. Outlined next are some of the questions concerning this phenomenon that led to the research efforts immediately following the initial discoveries, along with the answers, both complete and incomplete, obtained to those research questions. The basis for this first wave of follow-up research can be characterized by the phrase "what we knew we didn't know at that time." The final section is an update and brief overview of highlights of both "what we know now" and "what we now know that we don't know" about dynamic neuronal-astroglial interactions in the mHNS. PMID- 15134864 TI - Substance P in the baroreceptor reflex: 25 years. AB - Twenty-five years ago, very little was known about chemical communication in the afferent limb of the baroreceptor reflex arc. Subsequently, considerable anatomic and functional data exist to support a role for the tachykinin, substance P (SP), as a neuromodulator or neurotransmitter in baroreceptor afferent neurons. Substance P is synthesized and released from baroreceptor afferent neurons, and excitatory SP (NK1) receptors are activated by baroreceptive input to second order neurons. SP appears to play a role in modulating the gain of the baroreceptor reflex. However, questions remain about the specific role and significance of SP in mediating baroreceptor information to the central nervous system (CNS), the nature of its interaction with glutaminergic transmission, the relevance of colocalized agents, and complex effects that may result from mediation of non-baroreceptive signals to the CNS. PMID- 15134865 TI - Just cool it! Cryoprotectant anti-freeze in immunocytochemistry and in situ hybridization. AB - Immunohistochemical techniques offer specificity as well as flexibility for visualizing antigens. Their use with freely floating sections provides a high signal-to-noise ratio and has become a gold standard for brain and a number of other tissues. Yet this approach initially suffered from inability to keep the antigenicity in tissue sections and required immediate processing of all cut sections. Use of sucrose solutions enabled storage at refrigerator temperatures for a few days but longer-term storage was risky and either bacterial/fungal growth or evaporation of the storage solution compromised the integrity of the tissue. Our discovery 25 years ago that tissue sections can be stored for many years at -20 degrees C in an anti-freeze cryoprotectant solution with no loss of antigenicity solved this problem and has become widely used. More recently the utility of tissue stored for many years in anti-freeze cryoprotectant was pushed to new levels by testing new non-radioactive in situ hybridization (ISH) techniques that are based on modern immunocytochemistry. This review touches upon these advances in immunocytochemical technology using examples from neuroscience applications. PMID- 15134866 TI - Galanin in the brain: chemoarchitectonics and brain cartography--a historical review. AB - We present a review of galanin in the brain from a historical perspective of the development of "chemoarchitectonics" and "brain cartography" accomplished in the Histopharmacology Section at the National Institutes of Health. It was the mapping of potential brain neuroregulators that served as a springboard of ideas from which behavioral studies emanate. The integration of the known localization of neurotransmitter/neuromodulatory nerves ("chemoarchitectonic maps") and receptor binding sites with biochemical data derived from brain micropunches coupled with behavioral analysis at the level of discrete brain allows one to define the anatomical circuits which support behavioral changes and which ultimately will improve our understanding of mental disorders. PMID- 15134867 TI - NPY--an endearing journey in search of a neurochemical on/off switch for appetite, sex and reproduction. AB - Although a dynamic link between the two innate drives, appetite for food and the urge to reproduce, in vertebrate evolution has been known for a long time, a distinct neurochemical pathway mediating this integration has only recently been appreciated. Study of the precise anatomy of the neural track began in the early to mid 20th century after the sites of genesis of the two instincts were localized to the hypothalamus. This report narrates the birth and fruition to maturity of insights into the commonality of hypothalamic neuropeptide Y (NPY) signaling for the two instinctual drives along two distinct pathways. PMID- 15134868 TI - Hypothalamic control of energy balance: different peptides, different functions. AB - Energy balance is maintained via a homeostatic system involving both the brain and the periphery. A key component of this system is the hypothalamus. Over the past two decades, major advances have been made in identifying an increasing number of peptides within the hypothalamus that contribute to the process of energy homeostasis. Under stable conditions, equilibrium exists between anabolic peptides that stimulate feeding behavior, as well as decrease energy expenditure and lipid utilization in favor of fat storage, and catabolic peptides that attenuate food intake, while stimulating sympathetic nervous system (SNS) activity and restricting fat deposition by increasing lipid metabolism. The equilibrium between these neuropeptides is dynamic in nature. It shifts across the day-night cycle and from day to day and also in response to dietary challenges as well as peripheral energy stores. These shifts occur in close relation to circulating levels of the hormones, leptin, insulin, ghrelin and corticosterone, and also the nutrients, glucose and lipids. These circulating factors together with neural processes are primary signals relaying information regarding the availability of fuels needed for current cellular demand, in addition to the level of stored fuels needed for long-term use. Together, these signals have profound impact on the expression and production of neuropeptides that, in turn, initiate the appropriate anabolic or catabolic responses for restoring equilibrium. In this review, we summarize the evidence obtained on nine peptides in the hypothalamus that have emerged as key players in this process. Data from behavioral, physiological, pharmacological and genetic studies are described and consolidated in an attempt to formulate a clear statement on the underlying function of each of these peptides and also on how they work together to create and maintain energy homeostasis. PMID- 15134869 TI - Our journey with neuropeptide Y: effects on ingestive behaviors and energy expenditure. AB - Clark and colleagues first described the robust orexigenic effects of neuropeptide Y (NPY) in 1984. Our group as well as Stanley et al. confirmed these effects in the same year. During the next 20 years, we investigated the effects of NPY on diet preferences, opioid-related feeding, distributed neural feeding networks, energy metabolism, motivation and discriminative stimulus effects. These data together with data from other laboratories indicate that NPY increases feeding, even when rats work for food; that NPY decreases energy expenditure, particularly by altering thermogenesis; and that NPY's effects on energy metabolism are mediated by a widely distributed neural network involving other neuroregulators known to be involved in energy regulation. PMID- 15134870 TI - Bombesin-like peptides and associated receptors within the brain: distribution and behavioral implications. AB - As we commemorate the 25th anniversary of the journal Peptides, it is timely to review the functional significance of the bombesin (BB)-like peptides and receptors in the CNS. Over two decades ago we published an article in the journal Peptides demonstrating that BB-like peptides are present in high densities in certain rat brain regions (such as the paraventricular nucleus of the hypothalamus). Subsequently, one of the mammalian forms of BB, gastrin-releasing peptide (GRP) containing cell bodies were found in the suprachiasmatic nucleus of the hypothalamus and nucleus of the solitary tract of the hindbrain. Another related peptide, namely neuromedin (NM)B, was detected in the olfactory bulb and dentate gyrus. BB and GRP bind with high affinity to BB(2) receptors, whereas NMB binds with high affinity to BB(1) receptors. The actions of BB or GRP are blocked by BB(2) receptor antagonists such as (Psi(13,14)-Leu(14))BB whereas PD168368 is a BB(1) receptor antagonist. Exogenous administration of BB into the rat brain causes hypothermia, hyperglycemia, grooming and satiety. BB-like peptides activate the sympathetic nervous system and appear to modulate stress, fear and anxiety responses. GRP and NMB modulate distinct biological processes through discrete brain regions or circuits, and globally these peptidergic systems may serve in an integrative or homeostatic function. PMID- 15134871 TI - Angiotensin converting enzyme (ACE) and neprilysin hydrolyze neuropeptides: a brief history, the beginning and follow-ups to early studies. AB - Our investigations started when synthetic bradykinin became available and we could characterize two enzymes that cleaved it: kininase I or plasma carboxypeptidase N and kininase II, a peptidyl dipeptide hydrolase that we later found to be identical with the angiotensin I converting enzyme (ACE). When we noticed that ACE can cleave peptides without a free C-terminal carboxyl group (e.g., with a C-terminal nitrobenzylamine), we investigated inactivation of substance P, which has a C-terminal Met(11)-NH(2). The studies were extended to the hydrolysis of the neuropeptide, neurotensin and to compare hydrolysis of the same peptides by neprilysin (neutral endopeptidase 24.11, CD10, NEP). Our publication in 1984 dealt with ACE and NEP purified to homogeneity from human kidney. NEP cleaved substance P (SP) at Gln(6)-Phe(7), Phe(7)[see text]-Phe(8), and Gly(9)-Leu(10) and neurotensin (NT) at Pro(10)-Tyr(11) and Tyr(11)-Ile(12). Purified ACE also rapidly inactivated SP as measured in bioassay. HPLC analysis showed that ACE cleaved SP at Phe(8)-Gly(9) and Gly(9)-Leu(10) to release C terminal tri- and dipeptide (ratio = 4:1). The hydrolysis was Cl(-) dependent and inhibited by captopril. ACE released only dipeptide from SP free acid. ACE hydrolyzed NT at Tyr(11)-Ile(12) to release Ile(12)-Leu(13). Then peptide substrates were used to inhibit ACE hydrolyzing Fa-Phe-Gly-Gly and NEP cleaving Leu(5)-enkephalin. The K(i) values in microM were as follows: for ACE, bradykinin = 0.4, angiotensin I = 4, SP = 25, SP free acid = 2, NT = 14, and Met(5) enkephalin = 450, and for NEP, bradykinin = 162, angiotensin I = 36, SP = 190, NT = 39, Met(5)-enkephalin = 22. These studies showed that ACE and NEP, two enzymes widely distributed in the body, are involved in the metabolism of SP and NT. Below we briefly survey how NEP and ACE in two decades have gained the reputation as very important factors in health and disease. This is due to the discovery of more endogenous substrates of the enzymes and to the very broad and beneficial therapeutic applications of ACE inhibitors. PMID- 15134872 TI - Bradykinin antagonists: discovery and development. AB - Practical bradykinin antagonists were discovered in 1984 by Vavrek and Stewart and reported in "Peptides." At that time there was already much evidence for involvement of bradykinin in inflammation and pain, so the specific, competitive antagonists were widely accepted and applied. The key to conversion of bradykinin into an antagonist was replacement of the proline residue at position 7 with a D aromatic amino acid. Other modifications converted the initial weak antagonists into modern peptides which are totally resistant to all degrading enzymes, are orally available, and have been used in clinical trials. Non-peptide bradykinin antagonists have also been developed. PMID- 15134873 TI - Transport of radon gas into a tunnel at Yucca Mountain--estimating large-scale fractured tuff hydraulic properties and implications for the operation of the ventilation system. AB - Radon gas concentrations have been monitored as part of the operation of a tunnel (the Exploratory Studies Facility-ESF) at Yucca Mountain to ensure worker safety. The objective of this study was to examine the potential use of the radon data to estimate large-scale formation properties of fractured tuffs. This objective was examined by developing a numerical model, based upon the characteristics of the ESF and the Topopah Spring welded (TSw) tuff unit, capable of predicting radon concentrations for prescribed ventilation conditions. The model was used to address two specific issues. First, it was used to estimate the permeability and porosity of the fractures in the TSw at the length scale of the ESF and extending tens of meters into the TSw, which surrounds the ESF. Second, the model was used to understand the mechanism leading to radon concentrations exceeding a specified level within the ESF. The mechanism controlling radon concentrations in the ESF is a function of atmospheric barometric fluctuations being propagated down the ESF along with ventilated air flow and the slight suction induced by the ventilation exhaust fans at the South Portal of the ESF. These pressure fluctuations are dampened in the TSw fracture continuum according to its permeability and porosity. Consequently, as the barometric pressure in the ESF drops rapidly, formation gases from the TSw are pulled into the ESF, resulting in an increase in radon concentrations. Model calibration to both radon concentrations measured in the ESF and gas-phase pressure fluctuations in the TSw yielded concurrent estimates of TSw fracture permeability and porosity of 1 x 10( 11) m2 and 0.00034, respectively. The calibrated model was then used as a design tool to predict the effect of adjusting the current ventilation-system operation strategy for reducing the probability of radon gas concentrations exceeding a specified level. PMID- 15134874 TI - Microbial degradation of methyl tert-butyl ether and tert-butyl alcohol in the subsurface. AB - The fate of fuel oxygenates such as methyl tert-butyl ether (MTBE) in the subsurface is governed by their degradability under various redox conditions. The key intermediate in degradation of MTBE and ethyl tert-butyl ether (ETBE) is tert butyl alcohol (TBA) which was often found as accumulating intermediate or dead end product in lab studies using microcosms or isolated cell suspensions. This review discusses in detail the thermodynamics of the degradation processes utilizing various terminal electron acceptors, and the aerobic degradation pathways of MTBE and TBA. It summarizes the present knowledge on MTBE and TBA degradation gained from either microcosm or pure culture studies and emphasizes the potential of compound-specific isotope analysis (CSIA) for identification and quantification of degradation processes of slowly biodegradable pollutants such as MTBE and TBA. Microcosm studies demonstrated that MTBE and TBA may be biodegradable under oxic and nearly all anoxic conditions, although results of various studies are often contradictory, which suggests that site-specific conditions are important parameters. So far, TBA degradation has not been shown under methanogenic conditions and it is currently widely accepted that TBA is a recalcitrant dead-end product of MTBE under these conditions. Reliable in situ degradation rates for MTBE and TBA under various geochemical conditions are not yet available. Furthermore, degradation pathways under anoxic conditions have not yet been elucidated. All pure cultures capable of MTBE or TBA degradation isolated so far use oxygen as terminal electron acceptor. In general, compared with hydrocarbons present in gasoline, fuel oxygenates biodegrade much slower, if at all. The presence of MTBE and related compounds in groundwater therefore frequently limits the use of in situ biodegradation as remediation option at gasoline-contaminated sites. Though degradation of MTBE and TBA in field studies has been reported under oxic conditions, there is hardly any evidence of substantial degradation in the absence of oxygen. The increasing availability of field data from CSIA will foster our understanding and may even allow the quantification of degradation of these recalcitrant compounds. Such information will help to elucidate the crucial factors of site-specific biogeochemical conditions that govern the capability of intrinsic oxygenate degradation. PMID- 15134875 TI - Experimental study and modeling of the transfer of zinc in a low reactive sand column in the presence of acetate. AB - Nowadays, it is necessary to understand and identify the reactions governing the fate of heavy metals introduced into the environment with low complexing organic compounds, particularly when they are transferred through soils in urban areas. In this work the concomitant influence of pH and acetate on the fate of zinc on siliceous sand was studied in batch and non-saturated column experiments. Total zinc concentrations varied between 2 and 20 mg/l, and total acetate concentrations were fixed at 22, 72, 132, and 223 mM to obtain solution pHs of 4, 5, 6 and 7, respectively. Natural sand (diameter, 0.3-2 mm), mainly constituted of silica, was used. In batch adsorption experiments, zinc adsorption is insignificant at pH 4, low and linear at pH 5, and increasingly nonlinear, of the Langmuir type, at pH 6 and 7 indicating near-saturation conditions of surface sites at these high pH values. In column experiments, Zn retardation increases and the maximum outlet concentration of Zn decreases with rising pH and acetate concentrations. Previous column tracer experiments revealed the occurrence of regionalized water transport in the column. Modeling these data was based on a non-electrostatic approach. Batch and column data modeling was based on the PHREEQC code that allows concomitant resolution of chemical speciation and regionalized water transport. The speciation calculation indicates that the ZnAcetate+ species is the dominant Zn species in the solutions used. Batch experimental curves are correctly modeled assuming the formation of the three surface species triple bond SiOZn+, triple bond SiOH-Zn Acetate+ and triple bond SiO-Zn(Acetate)2-. The column data could be adequately modeled assuming a two region water transport and the formation of the same three species with the same thermodynamic constants determined in the batch experiments. The hypothesis of the modeling leads to a slight overestimation of the quantities of zinc eluted (10%) at pH 6 and 7, mostly in the desorption phase. These results show that the methodology used facilitates the correct modeling of both batch and transport experiments and formulation of the hypothesis on the interactions between the low reactive sand and a complex solution. PMID- 15134876 TI - A composite medium approximation for unsaturated flow in layered sediments. AB - Saturated-unsaturated flow in strictly layered sediments proceeds via conductors in parallel in the direction parallel to bedding, and via resistors in series in the direction perpendicular to bedding. On sufficiently small scales of space and time, flow in such media will be subject to approximate capillary equilibrium locally, which provides a basis for approximating the effective hydraulic conductivity of a composite multi-layer medium in terms of the conductivities of the individual layers. Equations for the hydraulic conductivity tensor in "composite medium approximation" (COMA) are given in a coordinate system aligned with bedding. Hydraulic conductivity parallel to bedding is generally larger than in the perpendicular direction. The anisotropy depends on the spread of the conductivity distribution, and tends to increase for dryer conditions. The COMA model was implemented in a multi-phase flow simulator and tested by comparison with high-resolution simulations in which all layering heterogeneity is resolved explicitly. Under favorable conditions, COMA is found to accurately represent sub grid scale flow and transport processes, providing a practical method for simulating field-scale flow and transport in layered media. The approximation improves when layers are thinner, and when flow rates are smaller. PMID- 15134877 TI - Reactive transport modelling of biogeochemical processes and carbon isotope geochemistry inside a landfill leachate plume. AB - The biogeochemical processes governing leachate attenuation inside a landfill leachate plume (Banisveld, the Netherlands) were revealed and quantified using the 1D reactive transport model PHREEQC-2. Biodegradation of dissolved organic carbon (DOC) was simulated assuming first-order oxidation of two DOC fractions with different reactivity, and was coupled to reductive dissolution of iron oxide. The following secondary geochemical processes were required in the model to match observations: kinetic precipitation of calcite and siderite, cation exchange, proton buffering and degassing. Rate constants for DOC oxidation and carbonate mineral precipitation were determined, and other model parameters were optimized using the nonlinear optimization program PEST by means of matching hydrochemical observations closely (pH, DIC, DOC, Na, K, Ca, Mg, NH4, Fe(II), SO4, Cl, CH4, saturation index of calcite and siderite). The modelling demonstrated the relevance and impact of various secondary geochemical processes on leachate plume evolution. Concomitant precipitation of siderite masked the act of iron reduction. Cation exchange resulted in release of Fe(II) from the pristine anaerobic aquifer to the leachate. Degassing, triggered by elevated CO2 pressures caused by carbonate precipitation and proton buffering at the front of the plume, explained the observed downstream decrease in methane concentration. Simulation of the carbon isotope geochemistry independently supported the proposed reaction network. PMID- 15134878 TI - In situ tracer tests to determine retention properties of a block scale fracture network in granitic rock at the Aspo Hard Rock Laboratory, Sweden. AB - Experiments were conducted at the Aspo Hard Rock Laboratory in order to improve the understanding of radionuclide retention properties of fractured crystalline bedrock in the 10-100 m scale (TRUE Block Scale Project, jointly funded by ANDRA, ENRESA, Nirex, JNC, Posiva and SKB). A series of tracer experiments were performed using sorbing tracers in three different flow paths. The different flow paths had Euclidian lengths of 14, 17 and 33 m, respectively, and one to three water conducting structures. Four tests were performed using different cocktails made up of radioactive sorbing tracers (22,24Na+, 42K+, 47Ca2+, 85Sr2+, 83,86Rb+, 131,133Ba2+ and 134,137Cs+). For each tracer injection, the breakthrough of sorbing tracers was compared to the breakthrough of a conservative tracer, 82Br-, 131I-, HTO and 186ReO4-, respectively. In the two longer flow paths, no breakthrough of 83Rb+ and 137Cs+ was observed after 8 months of pumping. Selected tracer tests were subject to basic modelling in which a one-dimensional (1D) advection-dispersion model, including surface sorption, and an unlimited matrix diffusion were used for the interpretation of the results. The results of the modelling indicated that there is a slightly higher mass transfer into a highly porous material in the block-scale experiment compared with in situ experiments performed over shorter distances and significantly higher than what would have been expected from laboratory data obtained from studies of the interactions in nonaltered intact rock. PMID- 15134879 TI - VEX-capture: a new technique that allows in vivo excision, cloning, and broad host-range transfer of large bacterial genomic DNA segments. AB - We have developed a novel and easily performed procedure for the targeted excision, cloning, and broad-host-range transfer of large bacterial genomic DNA segments. This procedure, called Vector-mediated excision and Capture (VEX Capture), represents a new molecular tool for the convenient manipulation and exchange of large (20-40+ kb) bacterial genomic fragments. VEX-Capture utilizes lox/Cre-mediated site-specific recombination for excision of the targeted genomic segment and homologous recombination for cloning of the excised DNA section onto a self-transmissible, broad-host-range IncP plasmid. The "captured" genomic DNA segment can then be transferred to a wide variety of Gram-negative hosts for basic research and bioengineering purposes. To demonstrate the utility and function of VEX-Capture, we have excised and cloned three separate genomic islands from the Salmonella typhimurium chromosome ranging in size from 26.7 to 40.0 kb. To test the ability of these islands to be established in different bacterial hosts, we transferred them to six other Gram-negative species and monitored their establishment via phenotypic and molecular analysis. RT-PCR was used to assay the expression of selected S. typhimurium island genes in the different species. This analysis led to the discovery that an island-encoded master regulator of S. typhimurium virulence functions is expressed in a species specific manner. Our results demonstrate the potential for VEX-Capture to be used as a convenient genetic technique for fundamental biological applications in a wide variety of bacterial species. PMID- 15134880 TI - Development and validation of evolutionary algorithm software as an optimization tool for biological and environmental applications. AB - A flexible, extendable tool for the optimization of (micro)biological processes and protocols using evolutionary algorithms was developed. It has been tested using three different theoretical optimization problems: 2 two-dimensional problems, one with three maxima and one with five maxima and a river autopurification optimization problem with boundary conditions. For each problem, different evolutionary parameter settings were used for the optimization. For each combination of evolutionary parameters, 15 generations were run 20 times. It has been shown that in all cases, the evolutionary algorithm gave rise to valuable results. Generally, the algorithms were able to detect the more stable sub-maximum even if there existed less stable maxima. The latter is, from a practical point of view, generally more desired. The most important factors influencing the convergence process were the parameter value randomization rate and distribution. The developed software, described in this work, is available for free. PMID- 15134881 TI - Multicenter evaluation of reverse line blot assay for detection of drug resistance in Mycobacterium tuberculosis clinical isolates. AB - A multicenter study was conducted with the objective to evaluate a reverse line blot (RLB) assay to detect resistance to rifampin (RIF), isoniazid (INH), streptomycin (STR), and ethambutol (EMB) in clinical isolates of Mycobacterium tuberculosis. Oligonucleotides specific for wild type and mutant (drug resistance linked) alleles of the selected codons in the genes rpoB, inhA, ahpC, rpsL, rrs, embB, were immobilized on a nylon membrane. The RLB assay conditions were optimized following analysis of DNA samples with known sequences of the targeted genes. For validation of the method at different geographical locations, the membranes were sent to seven laboratories in six countries representing the regions with high burdens of multudrug-resistant tuberculosis. The reproducibility of the assay for detection of rpoB genotypes was initially evaluated on a blinded set of twenty reference DNA samples with known allele types and overall concordant results were obtained. Further mutation analysis was performed by each laboratory on the local strains. Upon RLB analysis of 315 clinical isolates from different countries, 132 (85.2%) of 155 RIF-resistant and 28 (51.0%) of 55 EMB-resistant isolates were correctly identified, showing applicability of the assay when targeting the rpoB hot-spot region and embB306. Mutations in the inhA and ahpC promoter regions, conferring resistance to INH, were successfully identified in respectively 16.9% and 13.2% of INH-resistant strains. Likewise, mutations in rrs513 and rpsL88 that confer resistance to STR were identified in respectively 15.1% and 10.7% of STR-resistant strains. It should be mentioned that mutation analysis of the above targets usually requires rather costly DNA sequencing to which the proposed RLB assay presents rapid and inexpensive alternative. Furthermore, the proposed method requires the same simple equipment as that used for spoligotyping and permits simultaneous analysis of up to 40 samples. This technique is a first attempt to combine different targets in a single assay for prediction of antituberculosis drugs resistance. It is open to further development as it allows easy incorporation of new probes for detection of mutations in other genes associated with resistance to second-line (e.g., fluoroquinolones) and new antituberculosis compounds. PMID- 15134882 TI - Proteomic analysis of Acinetobacter lwoffii K24 by 2-D gel electrophoresis and electrospray ionization quadrupole-time of flight mass spectrometry. AB - The MS/MS analysis by Electrospray ionization quadrupole-time of flight mass spectrometry (ESI-Q-TOF MS) was applied to identify proteins in proteome analysis of bacteria whose genomes are not known. The protein identification by ESI-Q-TOF MS was performed sequentially by database search and then de novo sequencing using MS/MS spectra. Soil bacteria having unanalyzed genome, Acinetobacter lwoffii K24 is an aniline degrading bacterium. In this report, we present the results of a comparison between the proteome profile of A. lwoffii K24 cultured in aniline- or succinate-containing media. Protein analysis was performed using two-dimensional gel electrophoresis (2-DE) with pH 3-10 immobilized pH gradient (IPG) strips followed by ESI-Q-TOF MS. More than 780 protein spots were detected by 2-DE from the soluble proteome. Forty-eight of these proteins were expressed exclusively in aniline cultured bacteria, and 81 proteins increased and 162 proteins decreased in aniline-cultured versus succinate cultured A. lwoffii K24. Internal amino acid sequences of 43 major protein spots were successfully determined by ESI-Q-TOF MS to try to identify the bacterial proteins responding to aniline culture condition. Since the A. lwoffii K24 genome is not yet sequenced, many proteins were found to be hypothetical. Comparative proteome analysis of the insoluble protein fractions showed that one novel protein that was strongly induced by succinate-cultured A. lwoffii K24 was repressed under aniline culture conditions. These results suggest that comprehensive analysis of bacterial proteomes by 2-DE and amino acid sequence analysis by ESI-Q-TOF MS is useful for understanding induced novel proteins of biodegrading bacteria. PMID- 15134883 TI - Application of a rapid direct viable count method to deep-sea sediment bacteria. AB - For the first time, a Live/Dead (L/D) Bacterial Viability Kit (BacLight ) protocol was adapted to marine sediments and applied to deep-sea sediment samples to assess the viability (based on membrane integrity) of benthic bacterial communities. Following a transect of nine stations in the Fram Strait (Arctic Ocean), we observed a decrease of both bacterial viability and abundance with increasing water (1250-5600 m) and sediment depth (0-5 cm). Percentage of viable (and thus potentially active) cells ranged between 20-60% within the first and 10 40% within the fifth centimetre of sediment throughout the transect, esterase activity estimations (FDA) similarly varied from highest (13.3+/-5.4 nmol cm(-3) h(-1)) to lowest values below detection limit down the sediment column. Allowing for different bottom depths and vertical sediment sections, bacterial viability was significantly correlated with FDA estimations (p<0.001), indicating that viability assessed by BacLight staining is a good indicator for bacterial activity in deep-sea sediments. Comparisons between total L/D and DAPI counts not only indicated a complete bacterial cell coverage, but a better ability of BacLight staining to detect cells under low activity conditions. Time course experiments confirmed the need of a rapid method for viability measurements of deep-sea sediment bacteria, since changes in pressure and temperature conditions caused a decrease in bacterial viability of up to 50% within the first 48 h after sample retrieval. The Bacterial Viability Kit proved to be easy to handle and to provide rapid and reliable information. It's application to deep-sea samples in absence of pressure-retaining gears is very promising, as short staining exposure time is assumed to lessen profound adverse effects on bacterial metabolism due to decompression. PMID- 15134884 TI - Development of a real-time PCR method for quantification of the three genera Dehalobacter, Dehalococcoides, and Desulfitobacterium in microbial communities. AB - We developed standard curves based on plasmids containing a 16S rRNA gene of a member of one of the three genera Dehalobacter, Desulfitobacterium, and Dehalococcoides. A large difference in amplification efficiency between the standard curves was observed ranging from 1.5 to 2.0. The total eubacterial 16S rRNA gene copy number determined in a sample DNA by using eubacterial primers and the three standard curves led to differences in the estimated copy numbers of a factor up to 73. However, the amplification efficiencies for one specific standard curve were the same independent of the PCR primer pair used. This allowed the determination of the abundance of a population expressed as fractional number, hence, the percentage of genus-specific copy numbers within the total eubacterial 16S rRNA gene copy numbers. Determination of the fractional numbers in DNA mixtures of known composition showed the accuracy of this approach. The average difference in threshold value between two 10-fold dilutions of DNA of pure cultures, mixtures thereof and of environmental samples was 3.45+/-0.34, corresponding to an average almost optimal amplification efficiency of 1.95. This indicated that the low amplification efficiency of certain standard curves seemed to be mainly a problem of the plasmid DNA used and not of the 16S rRNA gene of the target genera. PMID- 15134885 TI - The filtration-acclimatization method for isolation of an important fraction of the not readily cultivable bacteria. AB - We developed a novel method, the filtration-acclimatization method (FAM), which enables the isolation and cultivation of an important fraction of the bacterial diversity, which is not cultivable by standard methods. The method consists of a filtration step, which removes most of the readily cultivable bacteria able to overgrow slowly growing bacteria, and an acclimatization procedure that provides a slow transition from the low environmental substrate concentrations to the high concentration of standard microbial media. So far, we isolated in total 65 strains from surface freshwater habitats by utilizing FAM. The isolates are affiliated with Actinobacteria, Alpha-, Betaproteobacteria, Bacteroidetes, and Spirochaeta. All isolates are pure cultures and form visible colonies on agar plates with high substrate concentrations. For further analysis, strains sharing more than a 97% 16S rRNA gene sequence similarity were grouped into one taxon. Based on sequence similarities, 88% of the obtained taxa can be considered to be undescribed species (<97% similarity to closest species). The highest similarity value of the taxa to the respective closest related species ranged from 87.7% to 99.8%, and was on average 94.5%. For comparison we isolated, by direct plating of water samples on a rich agar medium, a similar number of taxa. Amongst these taxa the percentage of taxa, which can be considered to be undescribed species, was only half of the percentage found for the taxa isolated by FAM. More importantly, it was amongst the taxa obtained by the standard method no taxon that was closer related to an uncultured bacterium than to an isolate, while 56% of the taxa isolated by FAM were closely related to uncultured bacteria. PMID- 15134886 TI - Evaluation of serological assays based on a novel excreted antigen preparation for the diagnosis of Cutaneous Leishmaniasis in Panama. AB - The objective of the present study was to determine the efficacy of prototype diagnostic serological assays for American Cutaneous Leishmaniasis (ACL) in Panama. As such, we prospectively sampled 100 cutaneous leishmaniasis case patients and tested their sera in two serological assays based upon novel soluble antigen preparations made from propagating the parasites in a protein-free, serum free media. Using serum and a Leishmania mexicana antigen preparation to sensitize plates, the assay correctly identified 89% of the case-patients. While using serum with an antigen preparation from Leishmania braziliensis, the assay correctly identified 71% of the patients. Concerning both test formats, performance was near equal in true positive and presumptive positive subsets demonstrating the improved sensitivity of these assays over reference methods of choice. Since the incidence of leishmaniasis in Panama has increased dramatically in the past 10 years, these assays may be useful in clinical and epidemiological studies and control programs. PMID- 15134888 TI - Universal primer PCR with DGGE for rapid detection of bacterial pathogens. AB - A universal primer PCR (UPPCR) combined with denaturing gradient gel electrophoresis (DGGE) was evaluated as a method permitting the rapid detection of pathogens. The results show that this method is efficient at amplifying the conserved regions of bacterial 16S rRNA genes with universal primers and can detect causative bacterial pathogens rapidly. Six species of bacteria from fisheries (Pseudomonas fluorescens, Vibrio anguillarum, Aeromonas hydrophila, Vibrio fluvialis, Providencia rettgeri and Aeromonas sobria) were examined. Our results indicate that the approach we undertook can be adopted not only for axenic bacterial populations but also for mixed communities as well. Furthermore, we were able to achieve the rapid detection of multiple bacteria a single in sample. In addition, UPPCR-DGGE was shown to be better than previously reported UPPCR-single-stranded conformation polymorphism (SSCP)-based methods for the rapid detection of bacterial pathogens. PMID- 15134887 TI - Quantification of a novel group of nitrate-reducing bacteria in the environment by real-time PCR. AB - Nitrate reduction is performed by phylogenetically diverse bacteria. Analysis of narG (alpha subunit of the membrane bound nitrate reductase) trees constructed using environmental sequences revealed a new cluster that is not related to narG gene from known nitrate-reducing bacteria. In this study, primers targeting this as yet uncultivated nitrate-reducing group were designed and used to develop a real-time SYBR(R) Green PCR assay. The assay was tested with clones from distinct nitrate-reducing groups and applied to various environmental samples. narG copy number was high ranging between 5.08x10(8) and 1.12x10(11) copies per gram of dry weight of environmental sample. Environmental real-time PCR products were cloned and sequenced. Data was used to generate a phylogenetic tree showing that all environmental products belonged to the target group. Moreover, 16S rDNA copy number was quantified in the different environments by real-time PCR using universal primers for Eubacteria. 16S rDNA copy number was similar or slightly higher than that of narG, between 7.12x10(9) and 1.14x10(11) copies per gram of dry weight of environmental sample. Therefore, the yet uncultivated nitrate reducing group targeted in this study seems to be numerically important in the environment, as revealed by narG high absolute and relative densities across various environments. Further analysis of the density of the nitrate-reducing community as a whole by real-time PCR may provide insights into the correlation between microbial density, diversity and activity. PMID- 15134889 TI - Fast screening method for detection of acyl-HSL-degrading soil isolates. AB - A reliable method was developed for screening of bacteria isolates capable of degrading acyl-HSLs, the signal molecules in quorum-sensing-mediated processes of many Proteobacteria. The microtiter assay was based on the use of a GFP-marked Escherichia coli strain, which fluoresces upon the presence of acyl-HSLs. Measurement of GFP fluorescence with a Molecular Imager FX scanner (fluorometer) detected isolates capable of degrading acyl-HSLs. The potential of this method was demonstrated by isolation of different bacteria from a potato rhizosphere able to inactivate synthetic and natural acyl HSLs produced by Pectobacterium carotovorum subsp. carotovorum (Pcc) (Erwinia carotovora subsp. carotovora (Ecc)). PMID- 15134890 TI - Construction of a nontypeable Haemophilus influenzae-specific ectopic delivery vector. AB - Complementation of chromosomal mutations in trans can introduce artifacts due to the number of episomal copies of the gene in question. One solution is to study the gene expressed at a single ectopic site in cis. We have designed and constructed a vector that allows homologous recombination into a gene encoding a frame-shifted IS1016-V6 protein in the Haemophilus influenzae Rd KW20 chromosome (HI1018). This site is the location of the > or = 35 kilobase capsule locus in encapsulated type b and d strains. This locus is not present in the nontypeable Rd KW20 strain, thus allowing ectopic expression of genes homologously recombined into HI1018 without polar effects. PMID- 15134891 TI - Regulation of lymphocyte adhesion and migration by the small GTPase Rap1 and its effector molecule, RAPL. AB - Dynamic regulation of integrin-mediated adhesion is central to lymphocyte trafficking and antigen recognition. The small GTPase Rap1 is a potent stimulator of leukocyte integrins through modulation of affinity and avidity. In addition, lymphocyte Rap1 has unique abilities to trigger cell polarization and enhance cell motility. These characteristics of Rap1 contribute to adhesive interactions with antigen-presenting cells (APC) and the vascular endothelium. In the process of elucidating the molecular mechanisms of Rap1-mediated integrin activation, we have identified a novel Rap1-binding molecule, regulator of adhesion and cell polarization enriched in lymphoid tissues (RAPL). RAPL is predominantly expressed in immune cells, and mediates Rap1-triggered integrin activation upon TCR engagement and chemokine stimulation. Importantly, Rap1/RAPL signaling cooperatively regulates cell polarization and integrin activation. The linkage between cell polarization and integrin activation through Rap1/RAPL signaling likely provides immune cells with their dynamic trafficking capability. PMID- 15134892 TI - Cross-reactive recognition of viral and self-peptides by a "public" T cell receptor expressed by cytotoxic T lymphocytes expanded in multiple unrelated individuals. PMID- 15134893 TI - Spare CD14 molecules on human monocytes enhance the sensitivity for low LPS concentrations. AB - Human monocytes express on their plasma membrane relatively large number of CD14 molecules, known to play a crucial role in the lipopolisaccharide (LPS)-mediated cellular activation. Indirect data (J. Biol. Chem. 270 (1995) 9904) suggest that not all of these CD14 molecules participate in LPS-signaling, but the importance of these spare receptors and the exact number of CD14 involved in activation upon different LPS-stimuli is not known. Using different concentrations of a blocking anti-CD14 monoclonal antibody (mAb 60bca) we created monocytes with graded amounts of CD14. The exact number of occupied and free receptors was quantitated by flow cytometry and special mAb-labeled standard beads. The number of free CD14 molecules per monocyte in the presence of 10, 3.33, 0.73, 0.25 and 0.041 microg/ml mAb was 0, 13,100, 49,300, 97,700 and 165,900. Stimulation of these partially blocked monocytes with 0.1, 1, 10 and 100 ng/ml ReLPS in the presence of 3% human serum revealed that already 13,100 and 97,700 CD14 molecules provided a maximal Tumor necrosis factor alpha (TNFalpha) mRNA response using 100 and 10 ng/ml ReLPS, while the activation totally depended on the number of available CD14 molecules in the case of 1 and 0.1 ng/ml ReLPS. Our data imply that the number of CD14 molecules available for LPS-binding influence the cellular response. In the presence of higher concentrations of LPS only fractions of CD14 participate in the cell activation, while the presence of the spare receptors enhance the sensitivity against lower LPS amounts. PMID- 15134894 TI - Generation of leukemia-specific T-helper type 1 cells applicable to human leukemia cell-therapy. AB - Leukemic dendritic cells (DC) were induced from the peripheral blood (PB) or bone marrow (BM) of leukemia patients by culture with (i) GM-CSF + IL-3 (neutral condition); (ii) GM-CSF + IL-3 + IL-12 + IFN-gamma (type 1-condition); or (iii) GM-CSF + IL-3 + IL-4 (type 2-condition). Although leukemic cells rapidly differentiated into adhesive leukemic DC in all culture conditions, type1 conditions were the most suitable for inducing leukemic DC expressing high levels of HLA and costimulatory molecules. Addition of IL-2 after 2 days of culture induced a preferential growth of minor T cell populations interacting with leukemic DC. In particular, IFN-gamma-producing CD4+ Th1 cells were efficiently expanded in type 1 culture conditions but nor in neutral or type 2-conditions. However, CD4+ T cells expanded in neutral conditions showed Th1-like functions if they were pulsed with IFN-gamma for 2 days before harvest. Such Th1 cells produced IFN-gamma and exhibited cytotoxicity in response to autologous leukemia cells. We further demonstrated that IFN-gamma production of leukemia-specific Th1 cells was blocked by anti-HLA-DR mAb. Thus, we established a novel culture system for inducing leukemia-specific Th1 cells. PMID- 15134895 TI - Transient suppression of IgG1 with IL-6 over-expression in immunized TCR transgenic mice. AB - Helper T cell-derived cytokines play a pivotal role in the production of antigen specific IgG antibody by B cells. In order to examine the in vivo effect of massive activation of helper T cells on the production of specific antibodies, ovalbumin (OVA)-specific TCR transgenic mice (OVA23-3) were immunized with OVA and serum levels of antigen-specific antibodies were measured. As a result, a great enhancement of antigen-specific IgM secretion and delay of IgG secretion were observed while the T cells produced sufficient Th2 cytokines. Immediately after OVA-immunization, marked increase of serum IL-6 was noted, which was followed by a transient increase of antibody forming cells. Both in vivo and in vitro experiments demonstrated that excess amount of IL-6 enhanced IgM production by activated B cells while suppressing IgG production. These results suggest that overproduction of IL-6 in the early stages of the primary immunization promotes development of IgM producing cells and causes the delay of IgG1 secretion. PMID- 15134896 TI - Regulation of perioperative immunological changes following laparotomy: effects of biological response modifier (BRM) on surgical stress. AB - Immune responses have been reported to decline following surgical stress, leading to an increased susceptibility to infection or to the growth of tumors. In this study, we report that pre-operative treatment with the biological response modifier (BRM) polysaccharide Kureha (PSK) can inhibit the decline of immunocompetence during the perioperative period. BALB/c mice were laparotomized, the intestinal tracts were exposed to room air for 1.5 h, and then the abdomens were closed. Six hours after the operation, the spleen and serum were collected. The concentrations of IL-6 in the serum and of IFNgamma and IL-4 in the supernatant of splenocyte cultures were measured. In the surgical stress group that received surgical stress and the PSK treatment, the serum IL-6 concentration was significantly elevated in the group with surgical stress. PSK treatment controlled the stress-induced elevation. IFNgamma concentrations were measured in the supernatant of Concanavalin A (Con A) stimulated splenocyte cultures. It was lower in the group with surgical stress than in the cultures derived from the non treated group. The IFNgamma concentration in the group with surgical stress plus PSK treatment was significantly higher than the level in the group with surgical stress alone. The IL-4 concentration was significantly lower in the surgical stress group than in the control group, however, the concentration tended to be higher in the surgical stress plus PSK treatment group than in the group with surgical stress alone. The IFNgamma/IL-4 ratio in the group with surgical stress was lower than the ratio in the non-treated group. The ratio in the group with surgical stress plus PSK treatment was significantly higher than the ratio in the group with surgical stress alone. These results suggest that PSK restores the abnormality of the biological responses induced by surgical stress and corrects the reduced Th1/Th2 cytokine balance to a normal level. PMID- 15134897 TI - Interleukin-17 augments tumor necrosis factor-alpha-induced elaboration of proangiogenic factors from fibroblasts. AB - Interleukin-17 (IL-17) is a CD4 T cell cytokine. In this report, we investigated the effects of this cytokine on the elaboration of proangiogenic factors by lung fibroblasts. After stimulation with a wide range of doses of IL-17, fibroblasts produced more amount of various kinds of angiogenic factors including NO, HGF, MCP-1, KC, MIP-2, PGE1, PGE2 and VEGF in a dose-dependent manner. Treatment with a COX-1 and COX-2 inhibitor indomethacin did not impair IL-17-induced HGF and VEGF secretion in fibroblasts. In addition, TNF-alpha alone stimulated the elaboration of KC, MIP-2, PGE2 and VEGF in fibroblasts. IL-17 and TNF-alpha in combination up-regulated elaboration of these proangiogenic factors additively or synergistically. Moreover, conditioned media (CM) from IL-17-stimulated fibroblasts showed significantly higher activity on endothelial cell growth than those from non-treated control cells. These results indicate that IL-17 up regulates elaboration of various proangiogenic factors, and modulates macrophage derived TNF-alpha-induced production of KC, MIP-2, PGE2 and VEGF by fibroblasts. Our findings also demonstrate that IL-17 might be a potential contributor to the inflammatory angiogenesis via induction of proangiogenic factors by stromal fibroblasts. PMID- 15134898 TI - The natural history of allergy: the development of new sensitizations in asthmatic children. AB - Allergy is an important risk factor for asthma in children as most of asthmatic children are sensitized. The aim of the study was to investigate the natural course of sensitizations in a group of children (340) at the onset of asthmatic symptoms. We subdivided children in three groups depending on the age at first visit (3-7-11 years) and followed them for 4 years. All children underwent three visits, each including skin prick test, every two years. In the 3-year age group (n = 133), 59% of children were sensitized at first visit, but the percentage increase to 88% at third visit. When looking only at allergic children at first visit (261), we demonstrated that polysensitization became higher than monosensitization since the age of 11. Analysis of sensitization distribution over the time revealed that house dust mites were prominent from the age of 3 years. Parietaria and grass pollens as well as olive pollen significantly increased, particularly during the 4-year follow-up in the 7-year group. Analysis of sensitization score demonstrated that the age range from 3 to 11 is at high risk to polysensitization. In conclusion, this study provides evidence that sensitization is frequent in asthmatic children, polysensitization represents the natural history of allergy and the type of immune response should appear different for each allergen. PMID- 15134899 TI - Mouse monoclonal IgA binds to the galectin-3/Mac-2 lectin from mouse macrophage cell lines. AB - The beta-galactoside-binding S-type lectin galectin-3/Mac-2, expressed among several other cell types on activated macrophages, is known to bind IgE, but not other Ig classes. We report in this paper that the single major constituent in a detergent lysate from the J774 mouse macrophage cell line bound to a mouse monoclonal IgA-affinity column. This fraction has been identified by mass spectrometry analysis as galectin-3. Binding of both mouse IgA and IgE to galectin-3 coated plates was inhibited by lactose and asialofetuin. Furthermore, three different monoclonal IgAs bound also to purified recombinant hamster galectin-3 coated plates in a concentration dependent manner. The potential functional significance of IgA binding to galectin-3 within macrophages and possibly other cell types is discussed. PMID- 15134900 TI - Isotype profiles of anti-gp160 antibodies from HIV-infected patients in plasma and culture supernatants. AB - We studied isotype profiles of anti-HIV antibodies (Ab) in HIV-1-infected African patients with high viral loads and major B cell dysfunction. We focussed on IgG1, IgG3, and IgA as these classes and subclasses tend to support neutralizing functions against HIV. Total IgG1, IgG3 and IgA were detected in the plasma of both HIV-1-infected and HIV-negative African individuals, but there was significantly more IgG3, a rare subclass, in HIV-1-infected patients (P < 0.05). Anti-HIV-gp160 specific antibodies were detected in sera from nearly all HIV-1 infected individuals tested, but not in HIV- individuals: 10/10, 9/10 and 8/10 individuals displayed specific IgG1, IgG3 and IgA, respectively. In the corresponding PBMC cultures carried out in the presence of IL-10 and IL-2, there was specific IgG1 and IgA in 5/10, and 3/10, respectively, but no IgG3 was detected. When HAART-treated European HIV-infected PBMC cultures were tested using the same protocol, specific IgG3 was detected in 4/10 cultures, and was unaffected by the addition of soluble CD40L molecules. The present study thus shows that, despite lymph node disorganization in HIV-infected drug-naive Africans, these individuals retain the ability to produce HIV-specific IgG1, IgG3 and IgA. However, the specific mechanisms controlling the selective production of IgG3, probably the most potent subclass and a potential target of immuno intervention, warrants further investigation. PMID- 15134901 TI - An IgM-kappa rat monoclonal antibody specific for the type 1 sphingosine 1 phosphate G protein-coupled receptor with antagonist and agonist activities. AB - Sphingosine 1-phosphate (S1P) type 1G protein-coupled receptors (S1P1 GPCRs) are specific high-affinity transducers for this lipid growth factor and cellular mediator. S1P1 GPCRs are widely-expressed and physiologically critical in the cardiovascular and immune systems. Functional rat monoclonal antibodies (MoAbs) have been generated against human S1P1 GPCRs expressed in rat null-cell transductants to provide bioavailable agents capable of stimulating or suppressing the S1P-S1P1 GPCR axis. The rat IgM-kappa anti-S1P1 GPCR MoAb designated 4B5.2 binds specifically to native human or mouse S1P1 GPCRs in cell membranes, but not to solubilized and denatured S1P1 GPCRs. Specific binding of 32P-S1P to cellular S1P1 GPCRs is not blocked by 4B5.2. T cell chemotactic responses to S1P and S1P suppression of T cell chemotaxis to chemokines both are inhibited selectively by 4B5.2. In contrast, generation of gamma-interferon by stimulated T cells is diminished by 4B5.2 as by S1P. T cell S1P1 GPCR-selective antagonist and agonist effects of 4B5.2 in vivo may alter immune responses as distinctively as the available poly-S1P GPCR-directed pharmacological agents, without the undesirable side-effects attributable to actions of these agents on other S1P GPCRs. PMID- 15134902 TI - Microparticulate beta-glucan upregulates the expression of B7.1, B7.2, B7-H1, but not B7-DC on cultured murine peritoneal macrophages. AB - Beta-1,3-(D)-glucan from a variety of biological sources has been shown to enhance both humoral and cellular immune responses to a variety of antigens, infectious agents, and tumors. Nevertheless, its mode of action has not been fully defined. We sought to determine whether a 1-2 microm diameter microparticulate form of beta-glucan (MG) from the yeast Saccharomyces cerevisiae could regulate expression of B7 family glycoproteins on resident peritoneal macrophages from BALB/c mice. We discovered that MG uregulated B7.2 mRNA expression and enhanced the surface membrane expression of B7.2 glycoprotein. Although B7.1 mRNA was not upregulated above constitutive levels, MG treatment enhanced B7.1 glycoprotein expression on the macrophages, albeit to a lesser extent than B7.2. At the same time, the gene and cell surface expression of B7 H1, a putative negative regulator of T cell activity, was also upregulated by MG. The expression of B7-DC, another B7 family molecule with negative regulatory activity, was not affected by incubation with MG. This study has demonstrated that a microparticulate form of beta-glucan can enhance B7 co-stimulatory molecule expression on macrophages, thereby enabling these antigen-presenting cells to deliver the second signal to T-lymphocytes that express CD28. In addition, because MG also induces the expression of B7-H1, it may enable macrophages to provide a concomitant downregulatory signal to T-lymphocytes expressing PD-1 or related receptors. PMID- 15134903 TI - Microarray analysis of differentially expressed genes in cells resistant to HIV 1. AB - We have previously isolated two matched transformed human T cell clones: one of which is resistant to HIV-1 replication and secretes an HIV-1 resistance factor(s) (HRF) and the second which retains the susceptibility of the parental cell line to HIV-1 infection. We employed cDNA arrays to investigate the spectrum of changes in cellular gene expression that correlate with the acquisition of HIV 1 resistance and the secretion of HRF. Using a tissue based immunology/hematology array, we identified 29 transcripts that were differentially expressed by HRF(+) and HRF(-) cells. HRF(+) cells showed a selective down-regulation of 11 genes involved in transcription, several of which are implicated in either susceptibility of cells to HIV-1 or the promotion of HIV-1 transcription itself. In the group of the up-regulated genes, three were linked directly to the cellular resistance to HIV-1. One of the cDNAs placed on the array, representing the hypothetical protein KIAA0117 hybridized only with poly A+ RNA probes derived from HRF(+) cells. The specific up-regulation of two genes, the transcription repressor (CTCF) and hypothetical protein KIAA0117 was confirmed by RT-PCR and Northern blot. The role of KIAA0117 transcript in the resistance to HIV-1 replication needs to be determined. PMID- 15134904 TI - Effect of nitric oxide in the differentiation of human monocytes to dendritic cells. AB - The aim of this work was to study the influence of nitric oxide (NO) in the differentiation of human monocytes to dendritic cells. Human monocytes from healthy donors were differentiated to immature dendritic cells in presence of GM CSF and IL-4. Maturation of dendritic cells was achieved with GM-CSF and TNF alpha. Nitric oxide donors (SIN-1, DEA-NO or DETA-NO) were added during differentiation of monocytes to dendritic cells and also during dendritic cells maturation. Immature dendritic cells showed a characteristic phenotype CD80+ CD1a+ HLA-DR+ CD86+ CD40+ CD14(low/-), different from adherent monocytes CD80- CD1a- HLA-DR+ CD86+ CD40- CD14++. The addition of SIN-1 the first day of monocyte differentiation reduced cell viability and increased the percentage of apoptotic immature dendritic cells. Peroxynitrite donor, SIN-1, produced more toxic effects than DEA-NO or DETA-NO. An increase in the subpopulation CD1a+ CD80+ HLADR+ of immature dendritic cells was observed when SIN-1 or DEA-NO, but not DETA-NO, was added at the beginning of monocyte culture. There was a significant reduction in the expression of TNF-alpha receptor of mature dendritic cells when SIN-1 and DEA NO were added together GM-CSF and TNF-alpha at the beginning of maturation. The presence of SIN-1, DEA-NO or DETA-NO in maturation induced an increase of CD83+ cells. These results suggest that nitric oxide affects differentiation and maturation of dendritic cells and this effect depends on the nitric oxide donor used. PMID- 15134905 TI - Continuous beds (monoliths): stationary phases for liquid chromatography formed using the hydrophobic interaction-based phase separation mechanism. AB - The pioneering research work published by Hjerten et al. [J. Chromatogr. 473 (1989) 273] in 1989 dealing with development and application of the continuous bed (monolithic) technique as an attractive alternative for the classical packed columns in chromatography, stimulated further investigations in this direction. The research data published since that time on the development and application of the continuous beds formed using hydrophobic interaction-based phase separation mechanism are reviewed. Some innovative species of the beds, such as polyrotaxane beds or nonparticulate restricted-access materials for direct analysis of the biological fluids in the capillary format are also discussed. Characteristic features and practical details of the continuous bed technique are revealed. Due to many advantages, the continuous bed technique became a competitor with the traditional packings in capillary or chip-based microanalysis. The importance of the continuous bed morphology on the chromatographic characteristics is shown. The applicability of modern microscopic analysis to evaluate the morphology of the continuous beds is demonstrated. PMID- 15134906 TI - Development of non-electrophoretic assay method for DNA ligases and its application to screening of chemical inhibitors of DNA ligase I. AB - A new rapid assay method for DNA ligases has been developed, which allows direct quantification of enzyme activity without using the traditional polyacrylamide gel electrophoretic technique. In this method, the 5'-biotinylated nicked duplex was used as a substrate for the ligase reaction, in which the 5'-end of the first oligonucleotide (19-mer) on the nicked strand is biotinylated and the second oligonucleotide (20-mer) on the same strand is labeled with radioactive 32P at the 5'-end. After ligation of the biotinylated 19-mer oligonucleotide into the second oligonucleotide with the reaction of DNA ligases, the biotinylated 19-mer oligonucleotide is converted into the radioactive 39-mer oligonucleotide. The ligase reaction products were heat-denatured to release both ligated and unligated biotinylated oligonucleotides. The biotinylated oligonucleotides were then captured on a streptavidin-coated microtiter plate and counted. The results obtained using this method correlated very well with those from the standard assay method using electrophoresis. Using this assay method, we were able to screen a chemical library and identify new DNA ligase inhibitors structurally related to resorcinol, which has growth inhibitory effects on the human breast cancer cell, MCF-7. The method described here is anticipated to be very useful for screening DNA ligase inhibitors from chemical libraries. PMID- 15134907 TI - Immobilization of catalase by entrapment of permeabilized yeast cells in hen egg white using glutaraldehyde. AB - An immobilized preparation of whole cell-based catalase was obtained by cross linking the yeast cells permeabilized with toluene in hen egg white using glutaraldehyde. Optimal preparations were obtained when cross-linking was carried out for 2 h at 4 degrees C. Immobilized cells could be reused for the removal of H2O2 from milk. PMID- 15134908 TI - Quantitatively investigating monomethoxypolyethylene glycol modification of protein by capillary electrophoresis. AB - Capillary electrophoresis (CE) was applied to study quantitatively protein modification with succinimidyl succinate-activated monomethoxypolyethylene glycol (MPEG-SS). The heterogeneous distribution of modified proteins and the average modification degree were determined by CE, and the latter met with the results from 2,4,6-trinitrobenzenesulfonic acid (TNBS) spectrometric assay. It was found that the optimal buffer pH for the modification was between pH 7.4 and 8.4, and the modification degree decreased when the modified sample was preserved in high pH solutions. The protein fractions attached with different number of polyethylene glycols (PEGs) were monitored along the process of protein modification. CE was proved to be efficient to evaluate quantitatively several factors of the protein modification, including the modifier/protein molar ratio, the stability of conjugates in different pH environments, and the time course of modification process. PMID- 15134909 TI - Electrosynthesis of imprinted polyacrylamide membranes for the stereospecific L histidine sensor and its characterization by AC impedance spectroscopy and piezoelectric quartz crystal technique. AB - L-histidine/D-histidine-imprinted membranes have been synthesized by electropolymerizing acrylamide onto Au electrodes or Au-coated quartz crystal electrodes in the presence of L-histidine/D-histidine. AC impedance spectroscopy and piezoelectric quartz crystal (PQC) technique were employed to verify the template effect of the membranes. The selectively rebinding of the enantiomers of histidine by their respective imprinted polymer membranes was confirmed. The result may lead to the construction of biomimetic sensors for the stereospecific determination of L-histidine and the chiral separation of histidine. PMID- 15134910 TI - Determination of depolarisation- and agonist-evoked calcium fluxes on skeletal muscle cells in primary culture. AB - Changes in intracellular calcium concentration ([Ca2+]i) evoked by prolonged depolarisation (120 mM KCl) or by the application of 15 mM caffeine were measured on skeletal muscle cells in primary culture. The extrusion rate (PVmax) of calcium from the myoplasm was determined, which in turn enabled the calculation of the calcium flux (Fl) underlying the measured calcium transients. PVmax was found to increase during differentiation, from 107 +/- 10 microM/s at the early myotube stage to 596 +/- 36 microM/s in secondary myotubes. This was paralleled by a decrease in resting [Ca2+]i from 99 +/- 4 to 51 +/- 2 nM. The depolarisation evoked Fl rose to peak and then ceased despite the continuous presence of KCl. In contrast, the caffeine-induced Fl showed a peak and a clear steady-level with a peak-to-steady ratio of 5.6 +/- 1.2. Removal of external calcium suppressed the depolarisation--induced flux by 88 +/- 5% indicating that both an influx and a release from the SR underlie the K(+)-evoked calcium transients. Subsequent applications of caffeine resulted in essentially identical fluxes indicating an efficient refilling of the internal stores. Moreover, if a depolarisation-induced calcium transient preceded the second caffeine-evoked release, the latter was significantly larger than the first suggesting that much of the calcium that entered was stored in the SR rather than extruded. PMID- 15134911 TI - Structural and spectroscopic studies of tripodal [MgL]2+ chelates containing only nitrogen donor atoms: alkaline earth metal complexes as potential drug delivery agents. AB - Several tripodal diimine ligands, tris(2-(2-thiazolyl)methyliminoethyl)amine, 2 Tatren, tris(2-(4-(5-methyl)imidazolyl)methyliminoethyl)amine, 5-Me-4-Imtren, tris(2-(4-imidazolyl)methyliminoethyl)amine, 4-Imtren, tris(2-(2 imidazolyl)methyliminoethyl)amine, 2-Imtren, and their Mg(2+) complexes were prepared and characterized. X-ray diffraction studies show that the Mg(2+) ions are six-coordinate, with three acyclic imine N atoms and three imidazolyl or thiazolyl N atoms coordinated with the general formula [Mg(L)](ClO(4))(2) (L=4 Imtren (1), 2-Imtren (2), 2-Tatren (3), and 5-Me-4-Imtren (4)). These complexes are chiral with both Delta and Lambda isomers present in the unit cell. (1)H NMR titrations reveal that complexes also form in solution and that the chirality is maintained. Variable temperature (1)H NMR reveals that the Delta and Lambda isomers interconvert in the intermediate to slow time scale. The interconversion rate slows with increasing pK(a) of the ligand heterocycle, suggesting that interconversion proceeds through a partially dissociated state. These complexes undergo trans-metallation by Zn(2+), indicating that their ligands can be released in a kinetically facile manner to form more stable metal ion complexes. PMID- 15134912 TI - Novel gadolinium(III) polyaminocarboxylate macrocyclic complexes as potential magnetic resonance imaging contrast agents. AB - Two novel Gd(III) complexes with functionalised polyaminocarboxylate macrocycles, 1,4,7-tris(carboxymethyl)-9,24-dioxo-14,19-dioxa-1,4,7,10,23- pentaazacyclododecane (L(1)) and 1,4,7-tris(carboxymethyl)-9,25-dioxo-14,17,20 trioxa-1,4,7,10,23- pentaazacyclotridecane (L(2)), were prepared in good yield. Their potential use as magnetic resonance imaging (MRI) contrast agents (CAs) was evaluated by investigating their relaxation behaviour as a function of pH, temperature and magnetic field strength. The 1/T(1) proton relaxivities at 20 MHz and 25 degrees C of GdL(1) (5.87 mM(-1) s(-1)) and GdL(2) (6.14 mM(-1) s(-1)) were found to be significantly higher than the clinically used Gd 1,4,7,10 tetraazacyclododecane-1,4,7,10-tetraacetic acid (Gd(DOTA)(-)) and Gd diethylenetriaminepentaethanoic acid (Gd(DTPA)(2-)). The complexes possess one water molecule in the inner coordination sphere whose mean residence lifetime was estimated to be 1.1 and 1.5 micros at 25 degrees C by variable temperature (VT) (17)O NMR spectroscopy. PMID- 15134913 TI - Comparison of anti-hyperglycemic effect amongst vanadium, molybdenum and other metal maltol complexes. AB - A wide variety of vanadium-containing complexes have been tested, both in vivo and in vitro, as possible therapeutic agents for the oral treatment of type 2 diabetes mellitus. None so far has surpassed bis(maltolato)oxovanadium(IV) (BMOV) for glucose- and lipid-lowering in an orally available formulation. Ligand choice is clearly an important factor in pharmacological efficacy of vanadium compounds as insulin enhancing agents. In this study, we kept the ligand and dose the same, varying instead the metal ion bound to the maltolato ligand in a series of binary complexes of neutral charge. A requirement for vanadyl ion as the metal ion of choice was apparent; no other metal ion tested served as a suitable substitute. Amongst [MoO(2)](2+), Co(II), Cu(II), Cr(III), and Zn(II), only [MoO(2)](2+) and Co(II) showed any hypoglycemic activity at the ED(50) dose for bis(maltolato)oxovanadium(IV), 0.6 mmolkg(-1) by oral gavage in streptozotocin (STZ)-diabetic rats within 72 h of administration of compound. PMID- 15134914 TI - Competition between lithium and magnesium ions for the G-protein transducin in the guanosine 5'-diphosphate bound conformation. AB - Li(+) is the most effective drug used to treat bipolar disorder; however, its exact mechanism of action has yet to be elucidated. One hypothesis is that Li(+) competes with Mg2+ for the Mg2+ binding sites on guanine-nucleotide binding proteins (G-proteins). Using 7Li T1 relaxation measurements and fluorescence spectroscopy with the Mg2+ fluorophore furaptra, we detected Li(+)/Mg(2+) competition in three preparations: the purified G-protein transducin (Gt), stripped rod outer segment membranes (SROS), and SROS with purified Gt reattached (ROS-T). When purified ROS-T, SROS or transducin were titrated with Li+ in the presence of fixed amounts of Mg(2+), the apparent Li(+) binding constant decreased due to Li(+)/Mg(2+) competition. Whereas for SROS the competition mechanism was monophasic, for G(t), the competition was biphasic, suggesting that in G(t), Li(+)/Mg(2+) competition occurred with different affinities for Mg(2+) in two types of Mg(2+) binding sites. Moreover, as [Li(+)] increased, the fluorescence excitation spectra of both ROS-T and G(t) were blue shifted, indicating an increase in free [Mg(2+)] compatible with Li(+) displacement of Mg(2+) from two low affinity Mg(2+) binding sites of G(t). G(t) release from ROS T membrane was also inhibited by Li(+) addition. In summary, we found evidence of Li(+)/Mg(2+) competition in G(t)-containing preparations. PMID- 15134915 TI - Novel polynuclear platinum adducts detected during the reactions of [Pt(Met S,N)Cl2] with gamma-glutathione and L-cysteine. AB - The reactions of platinum(II) complexes with thiol containing molecules are highly relevant to the mechanism of action of platinum-based drugs. This work presents the electrospray mass spectrometry (ESMS) and NMR results on the reactions of [Pt(l-MetH-S,N)Cl(2)] (l-MetH: l-methionine) with gamma-glutathione (GSH) and l-cysteine (l-Cys) at different pH and different molar ratios. Polymeric species such as [Pt(2)(micro-SG-S)(2)(Met-S,N)(2)], [Pt(3)(micro-SG S)(4)(Met-S,N)(2)], [Pt(4)(micro-SG-S)(6)(Met-S,N)(2)] and [Pt(5)(micro-SG S)(8)(Met-S,N)(2)] (l-Met: deprotonated l-methionine) were detected and were stable for long hours. For both reactions, the polymerization extent decreased with the increase of pH. For the reaction of l-Cys, only mononuclear complex [Pt(l-Met-S,N)(l-Cys-S,N)] was observed when pH>9. The observation and identification of polymeric (higher than binuclear) adducts of Pt(II)/GSH and Pt(II)/l-Cys appears to be unprecedented. PMID- 15134916 TI - Configurations of metallocyclams and relevance to anti-HIV activity. AB - Antiviral cyclam macrocycles block viral entry into cells by binding to the CXCR4 co-receptor. Cyclams bind transition metal ions strongly and can potentially form a range of trans (I-V) and cis configurations which may be recognised differently by co-receptor proteins. A survey of the CSD (crystallographic structural database) shows that the trans-III configuration is the most common in the solid state for complexes of cyclam itself. Other configurations can be induced by N substitution or ternary complexation and by interaction with solvents in solution. We report X-ray structures for the square-planar trans-III complexes [Pd(cyclam)]Cl(2).2MeOH and the C-C linked dimer [Pd(2)(2,2'-bi-(1,4,8,11 tetraazacyclotetradecane))](ClO4)(4), in which the planes of the two cyclam rings are close to perpendicular (100.1 degrees ), and for tetra-N-benzyl-cyclam and its 5-coordinate Ni(II) complex [Ni(Bz(4)-cyclam)Cl]Cl which has the unusual trans-I configuration. PMID- 15134917 TI - The effect of metal ions on the electrochemistry of the antitumor antibiotic streptonigrin. AB - The effect of transition metal ions on the electrochemistry of 6-methoxy-5,8 quinolinedione (L1), 7-amino-6-methoxy-5,8-quinolinedione (L2) and the antitumor antibiotic streptonigrin (SN) was studied. In 10% methanol/water, the one electron reduction of quinones L1 and L2 to the corresponding semiquinones is shifted to more positive potentials upon addition of one equivalent of Zn(II), Ni(II), Co(II) or Cd(II) and is consistent with formation of a 1:1 complex involving the quinone(N) and adjacent quinone(O). Similar results are observed for Cu(II) and Mn(II), but the redox chemistry is also complicated by metal-based redox chemistry. The addition of further equivalents of M(II) results in a number of different coordination and electrochemical processes including formation of 1:1 and 2:1 complexes of the quinone, semiquinone and dianion. Under similar conditions, the 1:1 SN 2,2'-bipyridyl metal complex undergoes a reversible one electron reduction to the semiquinone. The redox potential of the quinone in SN was shifted positive in the presence of the metal ions, but both the magnitude of the shift, and the relative influence of the metals was different to ligands L1 and L2. The changes in redox chemistry of SN compared with L1 and L2 are consistent with the formation of the 2,2-bipyridyl complexes in which there is weaker coordination to the quinone(O) in ring A of SN. These results suggest that in vivo, metal ions such as Zn(II), Cu(II) and Mn(II) facilitate the initial reduction of streptonigrin to the semiquinone by capturing the semiquinone after SN is reduced by biological reductants. PMID- 15134919 TI - Light-induced multistep oxidation of dinuclear manganese complexes for artificial photosynthesis. AB - Two dinuclear manganese complexes, [Mn(2)BPMP(mu-OAc)(2)].ClO(4) (1, where BPMP is the anion of 2,6-bis([N,N-di(2-pyridinemethyl)amino]methyl)-4-methylphenol) and [Mn(2)L(mu-OAc)(2)].ClO(4) (2, where L is the trianion of 2,6-bis([N-(2 hydroxy-3,5-di-tert-butylbenzyl)-N-(2-pyridinemethyl)amino]methyl)-4 methylphenol), undergo several oxidations by laser flash photolysis, using ruthenium(II)-tris-bipyridine (tris(2,2-bipyridyl)dichloro-ruthenium(II) hexahydrate) as photo-sensitizer and penta-amminechlorocobalt(III) chloride as external electron acceptor. In both complexes stepwise electron transfer was observed. In 1, four Mn-valence states from the initial Mn(2)(II,II) to the Mn(2)(III,IV) state are available. In 2, three oxidation steps are possible from the initial Mn(2)(III,III)state. The last step is accomplished in the Mn(2)(IV,IV) state, which results in a phenolate radical. For the first time we provide firm spectral evidence for formation of the first intermediate state, Mn(2)(II,III), in 1 during the stepwise light-induced oxidation. Observation of Mn(2)(II,III) is dependent on conditions that sustain the mu-acetato bridges in the complex, i.e., by forming Mn(2)(II,III) in dry acetonitrile, or by addition of high concentrations of acetate in aqueous solutions. We maintain that the presence of water is necessary for the transition to higher oxidation states, e.g., Mn(2)(III,III) and Mn(2)(III,IV) in 1, due to a bridging ligand exchange reaction which takes place in the Mn(2)(II,III) state in water solution. Water is also found to be necessary for reaching the Mn(2)(IV,IV) state in 2, which explains why this state was not reached by electrolysis in our earlier work (Eur. J. Inorg. Chem (2002) 2965). In 2, the extra coordinating oxygen atoms facilitate the stabilization of higher Mn valence states than in 1, resulting in formation of a stable Mn(2)(IV,IV) without disintegration of 2. In addition, further oxidation of 2, led to the formation of a phenolate radical (g = 2.0046) due to ligand oxidation. Its spectral width (8 mT) and very fast relaxation at 15 K indicates that this radical is magnetically coupled to the Mn(2)(IV,IV) center. PMID- 15134918 TI - Cobalt(II) and zinc(II) binding to a ferredoxin maquette. AB - We have examined the Co(II) and Zn(II) affinity of the prototype ferredoxin maquette ligand, NH(2)-KLCEGG.CIACGAC.GGW-CONH2 (IAA), which was originally designed to bind a [4Fe-4S] cluster. UV-Vis spectroscopy demonstrates tight 1:1 complex formation between Co(II) and IAA. The intensity of the S-->Co(II) charge transfer bands at 304 and 340 nm and the ligand field bands between 630 and 728 nm indicate Co(II) coordination by the four cysteine thiolates of IAA in a pseudo tetrahedral geometry. A dissociation constant value of 5.3 microM was determined for the Co(II)-IAA complex at pH 6.5. Zn(II) readily displaces Co(II) from IAA as evinced by loss of the Co(II) spectral features. The dissociation constant for Zn(II), 20 pM at pH 6.5, was determined be competition experiments with Co(II) IAA. These results demonstrate that the ferredoxin maquette ligand is an excellent ligand for Zn(II). PMID- 15134920 TI - Dinuclear copper-dioxygen intermediates supported by polyamine ligands. AB - Reactivity of the dicopper(I) and dicopper(II) complexes supported by novel polyamine ligands L1 (1,11-bis(6-methylpyridin-2-yl)-2,6,10-triaza-2,6,10 tribenzylundecane) and L2 (5-benzyl-1,9-bis(6-methylpyridin-2-yl)-2,8-bis(6 methylpyridin-2-ylmethyl)-2,5,8-triazanonane) towards O(2) and H(2)O(2), respectively, has been investigated in order to shed light on the ligand effects on Cu(2)/O(2) chemistry. The dicopper(I) complex of L1 (1a) readily reacted with O(2) in a 2:1 ratio at a low temperature (-94 degrees C) in acetone to afford a mixture of (mu-eta2.eta2-peroxo)dicopper(II) and bis(mu-oxo)dicopper(III) complexes. The formation of these species has been confirmed by the electron spin resonance (ESR) silence of the solution as well as their characteristic absorption bands in the UV-visible region (gammamax= 350 and 510 nm due to the peroxo complex and approximately 400 nm due to the bis(mu-oxo) complex] and the resonance Raman bands at 729 cm(-1) [Deltanu (16(O2)-18(O2)) = 38 cm(-1)] due to the peroxo complex and at 611 and 571 cm(-1) [Deltanu(16(O2)-18(O2)) = 22 and 7 cm(-1), respectively] due to the bis(mu-oxo) complex. The peroxo and bis(mu-oxo) complexes were unstable even at the low temperature, leading to oxidative N dealkylation at the ligand framework. The dicopper(I) complex of L2 (2a) also reacted with O(2) to give (mu-hydroxo)dicopper(II) complex (2b(OH)) as the product. In this case, however, no active oxygen intermediate was detected even at the low temperature (-94 degrees C). With respect to the copper(II) complexes, treatment of the (mu-hydroxo)dicopper(II) complex of L1 (1b(OH)) with an equimolar amount of H(2)O(2) in acetone at -80 degrees C efficiently gave a (mu 1,1-hydroperoxo)dicopper(II) complex, the formation of which has been supported by its ESR-silence as well as UV-vis (370 and 650 nm) and resonance Raman spectra [881 cm(-1); [Deltanu (16(O2)-18(O2)) = 49 cm(-1)]. The (mu-1,1 hydroperoxo)dicopper(II) intermediate of L1 also decomposed slowly at the low temperature to give similar oxidative N-dealkylation products. Kinetic studies on the oxidative N-dealkylation reactions have been performed to provide insight into the reactivity of the active oxygen intermediates. PMID- 15134921 TI - Structural models for the reduced form of vanadate-dependent peroxidases: vanadyl complexes with bidentate chiral Schiff base ligands. AB - The penta-coordinated vanadyl complexes [VO(ON)(2)] have been obtained by reaction between [VOX(2)] (X = acetylacetonate or chloride) and the Schiff base ligands HON = (R)-sal-am, (R)-Clsal-am and (S)-naph-am, where sal and naph are the salicylidene and naphthalidene moieties, and am derives from phenylethylamine. The three complexes and the ligand (R)-Clsal-am have been structurally characterized. The geometry of the complexes is in-between trigonal bipyramidal (with the two imine functions in the axis) and square-pyramidal; tau values range from 0.66 to 0.44. Structural and EPR (electron paramagnetic resonance) features are in accord with the coordination environment proposed for the inactive, reduced (V(IV)) form of the bromoperoxidase from the marine brown alga Ascophyllum nodosum. PMID- 15134922 TI - Copper transfer to the N-terminal domain of the Wilson disease protein (ATP7B): X ray absorption spectroscopy of reconstituted and chaperone-loaded metal binding domains and their interaction with exogenous ligands. AB - The copper-transporting ATPases are 165-175 kDa membrane proteins, composed of 8 transmembrane segments and two large cytosolic domains, the N-terminal copper binding domain and the catalytic ATP-hydrolyzing domain. In ATP7B, the Wilson disease protein, the N-terminal domain is made up of six metal-binding sub domains containing the MXCXXC motif which is known to coordinate copper via the two cysteine residues. We have expressed the N-terminal domain of ATP7B as a soluble C-terminal fusion with the maltose binding protein. This expression system produces a protein which can be reconstituted with copper without recourse to the harsh denaturing conditions or low pH reported by other laboratories. Here we describe the reconstitution of the metal binding domains (MBD) with Cu(I) using a number of different protocols, including copper loading via the chaperone, Atox1. X-ray absorption spectra have been obtained on all these derivatives, and their ability to bind exogenous ligands has been assessed. The results establish that the metal-binding domains bind Cu(I) predominantly in a bis cysteinate environment, and are able to bind exogenous ligands such as DTT in a similar fashion to Atox1. We have further observed that exogenous ligand binding induces the formation of a Cu-Cu interaction which may signal a conformational change of the N-terminal domain. PMID- 15134924 TI - Displacement of iron by zinc at the diiron site of Desulfovibrio vulgaris rubrerythrin: X-ray crystal structure and anomalous scattering analysis. AB - X-ray crystal structures of recombinant Desulfovibrio (D.) vulgaris rubrerythrin (Rbr) have shown a diiron site, whereas the crystal structure of Rbr "as isolated" from D. vulgaris was reported to contain a mixed Zn,Fe binuclear site. To investigate the possibility that zinc had displaced iron during isolation or crystallization of the "as-isolated" D. vulgaris Rbr, the X-ray crystal structure of recombinant D. vulgaris all-iron Rbr that had been incubated with excess zinc sulfate prior to crystallization, yielding a protein labeled Zn,FeRbr, was solved. Analysis of the anomalous scattering data obtained at two different wavelengths showed that zinc had displaced a significant proportion of iron from both iron centers of the diiron site, and that no iron had been displaced from the [Fe(SCys)(4)] site. UV-visible absorption spectra of the redissolved Zn,FeRbr crystals showed 30-40% retention of oxo-bridged diferric sites, and the redissolved crystals had 37% of the peroxidase specific activity of the starting all-iron Rbr, which, together with the crystallographic results, indicate a predominant mixture of Fe1,Fe2 and Zn1,Zn2 sites. The structure of the Zn(Fe)1,Fe(Zn)2 binuclear site in the Zn,FeRbr crystals was very similar to that of the Zn,Fe binuclear site reported for the "as-isolated" D. vulgaris Rbr, including tetrahedral four-coordination at the Zn(Fe)1 site. The diiron sites in the recombinant Zn,FeRbr crystals were likely at least partially reduced during synchrotron irradiation. Our results suggest that the mixed-metal binuclear site reported for the "as-isolated" D. vulgaris Rbr could be due to displacement of iron from a native diiron site by adventitious zinc during isolation and/or crystallization, and that reduced diiron and dizinc sites can adopt very similar structures in Rbr. PMID- 15134925 TI - Spectroscopic evidence for interactions between hexacyanoiron(II/III) and an engineered purple CuA azurin. AB - Interactions between hexacyanoiron(II/III) and a dinuclear, mixed valence Cu(A) center in engineered Cu(A) azurin have been investigated by UV-visible (UV-vis) and electron paramagnetic resonance (EPR) spectroscopic techniques. Addition of ferricyanide (hexacyanoiron(III)) to the Cu(A) azurin resulted in a new absorption band around 500 nm in the UV-vis and an isotropic line at g = 2.16 in the EPR spectra. Control experiments, including additions of Cu(II)SO(4) or Cu(I)(CH(3)CN)(4)PF(6) to ferricyanide or ferrocyanide, as well as gel filtration purification of the ferricyanide-Cu(A) azurin adduct indicate complex formation between cupric ion and ferrocyanide ion in the protein. Solvent or small molecule accessibility, metal oxidation state and the presence of more than one metal ion are potential factors important for the complex formation. These findings must be taken into consideration when using ferricyanide or ferrocyanide as redox agents for studying Cu(A) centers in proteins. PMID- 15134923 TI - Zn-, Cd-, and Pb-transcription factor IIIA: properties, DNA binding, and comparison with TFIIIA-finger 3 metal complexes. AB - Properties of the metal ion binding sites of Zn-transcription factor IIIA (TFIIIA) were investigated to understand the potential of this type of zinc finger to undergo reactions that remove Zn(2+) from the protein. Zn-TFIIIA was purified from E. coli containing the cloned sequence for Xenopus laevis oocyte TFIIIA and its stoichiometry of bound Zn(2+) was shown to depend on the details of the isolation process. The average dissociation constant of Zn(2+) in Zn TFIIIIA was 10(-7). The dissociation constant for Zn-F3, the third finger from the N-terminus of TFIIIA, was 1.0 x 10(-8). The reactivity of Zn-TFIIIA with a series of metal binding ligands, including 2-carboxy-2'-hydroxy-5' sulfoformazylbenzene (zincon), 4-(2-pyridylazo)-resorcinol (PAR), and 3-ethoxy-2 oxo-butyraldehyde-bis-(N(4)-dimethylthiosemicarbazone) (H(2)KTSM(2)) revealed similar kinetics. The reactivity of PAR with Zn-TFIIIA declined substantially when the protein was bound to the internal control region (ICR) of the 5S ribosomal DNA. Both Cd(2+) and Pb(2+) disrupt TFIIIA binding to its cognate DNA sequence. The Pb(2+) dissociation constant of Pb-F3 was measured as 2.5 x 10(-8). According to NMR spectroscopy, F3 does not fold into a regular conformation in the presence of Pb(2+). PMID- 15134926 TI - Nickel trafficking: insights into the fold and function of UreE, a urease metallochaperone. AB - UreE is a metallo-chaperone assisting the incorporation of two adjacent Ni(2+) ions in the active site of urease. This study describes an attempt to distill general information on this protein using a computational post-genomic approach for the understanding of the structural details of the molecular function of UreE in nickel trafficking. The two crystal structures recently determined for UreE from Bacillus pasteurii (BpUreE) and Klebsiella aerogenes (KaUreE) were comparatively analyzed. This analysis provided insights into the protein structural and conformational features. A structural database of UreE proteins from a large number of different genomes was built using homology modeling. All available sequences of UreE were retrieved from protein and cDNA databases, and their structures were modeled on the crystal structures of BpUreE and KaUreE. A self-consistent iterative protocol was devised for multiple sequence alignment optimization involving secondary structure prediction and evaluation of the energy features of the obtained modeled structures. The quality of all models was tested using standard assessment procedures. The final optimized structure-based multiple alignment and the derived model structures provided insightful information on the evolutionary conservation of key residues in the protein sequence and surface patches presumably involved in protein recognition during the urease active site assembly. PMID- 15134928 TI - Resonance Raman study on synergistic activation of soluble guanylate cyclase by imidazole, YC-1 and GTP. AB - Soluble guanylate cyclase (sGC), a physiological nitric oxide (NO) receptor, is a heme-containing protein and catalyzes the conversion of GTP to cyclic GMP. We found that 200 mM imidazole moderately activated sGC in the coexistence with 3 (5'-hydroxymethyl-2'-furyl)-1-benzylindazole (YC-1), although imidazole or YC-1 alone had little effect for activation. GTP facilitated this process. Resonance Raman spectra of imidazole complex of native sGC and CO-bound sGC (CO-sGC) have demonstrated that a simple heme adduct with imidazole at the sixth coordination position is not present for both sGC and CO-sGC below 200 mM of the imidazole concentration and that the Fe-CO stretching band (nuFe-CO)) appears at 492 cm(-1) in the presence of imidazole compared with 473 cm(-1) in its absence. Both frequencies fall on the line of His-coordinated heme proteins in the nuFe-CO vs nuC-O plot. However, it is stressed that the CO-heme of sGC becomes apparently photo-inert in a spinning cell in the presence of imidazole, suggesting the formation of five-coordinate CO-heme or of six-coordinate heme with a very weak trans ligand. These observations suggest that imidazole alters not only the polarity of heme pocket but also the coordination structure at the fifth coordination side presumably by perturbing the heme-protein interactions at propionic side chains. Despite the fact that the isolated sGC stays in the reduced state and is not oxidized by O(2), sGC under the high concentration of imidazole (1.2 M) yielded nu4 at 1373 cm(-1) even after its removal by gel filtration, but addition of dithionite gave the strong nu4 band at 1360 cm(-1). This indicated that imidazole caused autoxidation of sGC. PMID- 15134927 TI - The stability of the cytochrome c scaffold as revealed by NMR spectroscopy. AB - NMR spectroscopy was used to study the effect of guanidinium chloride on the unfolding of horse heart and yeast iso-1 cytochrome c under mild alkaline conditions. The structural changes on the horse heart protein were detected through NOESY (Nuclear Overhauser Effect SpectroscopY) experiments whereas (15)N (1)H heteronuclear NMR was used to monitor the behavior of the yeast protein. The latter represents the first characterization through (15)N-(1)H heteronuclear NMR spectroscopy of the guanidinium chloride induced unfolding of mitochondrial cytochrome c. The presence of denaturants decreases the temperature at which the native Met80 axial ligand is displaced from the iron center under the present mild alkaline conditions. The process can be described in terms of protein fragments behaving as unfolding units of different stability. The comparison between the two proteins indicates that the loop+helix connecting the proximal and distal sites, as well as the long Met80-containing loop immediately after a short helix, are structural characteristics of mitochondrial cytochrome c that appear to be responsible for the Met80-iron(III) bond fragility. PMID- 15134929 TI - Antagonists Mo and Cu in a heterometallic cluster present on a novel protein (orange protein) isolated from Desulfovibrio gigas. AB - An orange-coloured protein (ORP) isolated from Desulfovibrio gigas, a sulphate reducer, has been previously shown by extended X-ray absorption fine structure (EXAFS) to contain a novel mixed-metal sulphide cluster of the type [S(2)MoS(2)CuS(2)MoS(2)] [J. Am. Chem. Soc. 122 (2000) 8321]. We report here the purification and the biochemical/spectroscopic characterisation of this novel protein. ORP is a soluble monomeric protein (11.8 kDa). The cluster is non covalently bound to the polypeptide chain. The presence of a MoS(4)(2-) moiety in the structure of the cofactor contributes with a quite characteristic UV-Vis spectra, exhibiting an orange colour, with intense absorption peaks at 480 and 338 nm. Pure ORP reveals an Abs(480)/Abs(338) ratio of 0.535. The gene sequence coding for ORP as well as the amino acid sequence was determined. The putative biological function of ORP is discussed. PMID- 15134930 TI - Studies on the mechanism of action of xanthine oxidase. AB - Recent studies of the reaction mechanism of the molybdenum-containing enzyme xanthine oxidase are presented. The pH-dependence of both the steady-state and rapid reaction kinetics of the enzyme exhibits is bell-shaped, with pK(a)s for the acid and alkaline limbs of 6.6 and 7.4, respectively. These are assigned to ionizations of an active site base and substrate, respectively, with the implication that enzyme acts on the neutral rather than monoanionic form of the purine substrate. A computational study provides evidence that in the course of the reaction tautomerization of substrate occurs, with a proton moving from N-3 to N-9 in the course of the reaction - enzyme facilitation of this tautomerization may contribute as much as 24 kcal/mol in transition state stabilization for the reaction. Electron spin echo (ESEEM) and electron-nuclear double resonance (ENDOR) studies of the so-called "very rapid" Mo(V) intermediate of the reaction, the latter work using a newly synthesized form of the substrate 2-hydroxy-6-methylpurine that has been selectively isotopically labeled at C-8, indicates that product is bound to the molybdenum of the active site in a simple, end-on fashion, consistent with a reaction mechanism involving nucleophilic attack of a (deprotonated) Mo-OH on the C-8 position of substrate. A kinetic study using a series of purines has failed to identify a correlation between the one-electron reduction potential for substrate and catalytic effectiveness, indicating that a reaction mechanism initiated by one-electron, outer-sphere electron transfer is unlikely. Finally, a consideration of the active site structure in the context of the above work suggests specific amino acid residues to target for site-directed mutagenesis studies. Preliminary experiments with two such mutants are entirely consistent with the proposed catalytic roles of two active site glutamate residues. PMID- 15134931 TI - Interaction of plastocyanin with oligopeptides: effect of lysine distribution within the peptide. AB - We synthesized and purified four oligopeptides containing four lysines (KKKK, GKKGGKK, KKGGGKK, and KGKGKGK) as models for the plastocyanin (PC) interacting site of cytochrome f. These peptides competitively inhibited electron transfer between cytochrome c and PC. The inhibitory effect increased as the peptide concentrations were increased. The association constants between PC and the peptides did not differ significantly (3500-5100 M(-1)), although the association constant of PC-KGKGKGK was a little larger than the constants between PC and other peptides. Changes in the absorption spectrum of PC were observed when the peptides were added to the PC solution: peaks and troughs were detected at about 460 and 630 nm and at about 560 and 700 nm, respectively, in the difference absorption spectra between the spectra with and without peptides. These changes were attributed to the structural change at the copper site of PC by interaction with the peptides. The structural change was most significant when tetralysine was used. These results show that binding of the oligopeptide to PC is slightly more efficient when lysines are distributed uniformly within the peptide, whereas the structural change of PC becomes larger when the lysines are close to each other within the peptide. PMID- 15134932 TI - Aberrant activity of the DNA repair enzyme AlkB. AB - Escherichia coli AlkB is a DNA/RNA repair enzyme containing a mononuclear Fe(II) site that couples the oxidative decomposition of alpha-ketoglutarate (alphaKG) to the hydroxylation of 1-methyladenine or 3-methylcytosine lesions in DNA or RNA, resulting in release of formaldehyde and restoration of the normal bases. In the presence of Fe(II), alphaKG, and oxygen, but the absence of methylated DNA, AlkB was found to catalyze an aberrant reaction that generates a blue chromophore. The color is proposed to derive from Fe(III) coordinated by a hydroxytryptophan at position 178 as revealed by mass spectrometric analysis. Protein structural modeling confirms that Trp 178 is reasonably positioned to react with the Fe(IV) oxo intermediate proposed to form at the active site. PMID- 15134933 TI - Relativistic DFT calculation of the reaction cycle intermediates of [NiFe] hydrogenase: a contribution to understanding the enzymatic mechanism. AB - Structures and spectroscopic observables of the paramagnetic intermediates of the enzymatic reaction cycle of the metalloenzyme [NiFe] hydrogenase were calculated using relativistic density functional theory (DFT) within the zero-order regular approximation (ZORA). By comparing experimental and calculated magnetic resonance parameters (g- and hyperfine tensors) for the states Ni-A, Ni-B, Ni-C, Ni-L, and Ni-CO the details of the atomic composition of these paramagnetic intermediates could be elucidated that are mostly not available from X-ray structure analysis. In general, good agreement between calculated and experimental observables could be obtained. A detailed picture of the changes of the active center during the catalytic cycle was deduced from the obtained structures. Based on these results, a consistent model for the sequence of redox states including protonation steps is proposed which is important for understanding the mechanism of the [NiFe] hydrogenase. PMID- 15134934 TI - The structure of sitting-atop complexes of metalloporphyrins studied by theoretical methods. AB - The metallation of tetrapyrroles is believed to proceed via a sitting-atop (SAT) complex, in which some of the pyrrole nitrogen atoms are still protonated and the metal ion resides above the ring plane. No crystal structure of such a complex has been presented, but NMR and extended X-ray absorption fine structure (EXAFS) data has been reported for Cu(2+) in acetonitrile. We have used density functional calculations to obtain reasonable models for SAT complexes of porphyrins with Mg(2+), Fe(2+), and Cu(2+). The results show that there are many possible SAT complexes with 1-5 solvent molecules, one or two metal ions, and cis or trans protonation of the porphyrin ring. Many of these have similar energies and their relative stabilities vary with the metal ion. A complex with two cis pyrrolenine nitrogens atoms and 2-4 solvent molecules coordinated to Cu(2+) fits the NMR and EXAFS data best. However, we cannot fully exclude the possibility that what is observed is rather a mixture of a doubly protonated porphyrin and the copper porphyrin. Mg(2+) has a lower affinity for porphyrin and stronger affinity for water, so a complex with five water molecules and only one bond to porphyrin seems to be most stable. For Fe(2+), a cis structure with two first sphere water molecules and four interactions to the porphyrin seems to be most likely. PMID- 15134935 TI - Interpretation of the temperature-dependent color of blue copper protein mutants. AB - The electronic absorption spectrum of the mutant of the blue copper protein amicyanin with a pseudoazurin loop (AmiPse) shows a remarkable temperature dependence. The absorption band at approximately 460 nm increases at low temperature while the transition at approximately 600 nm is not much affected by a variation of the temperature. An approximate density functional theory (DFT) study of the active site model [Cu(II)(imidazole)(2)(SCH(3))(S(CH(3))(2))](+) (protein backbone and solvation neglected) leads to two local minimum structures (axial and rhomb) which both have a geometry close to that typical for blue copper proteins. One (rhomb) has two structurally different histidine donors, and this geometry is also found in most experimental type 1 structures. The two forms axial and rhomb are distortional isomers and are energetically almost degenerate. The temperature dependence of the spectrum of AmiPse is interpreted with a temperature-dependent change of the relative population of the two local minimum structures with slightly different energy. The 460 nm transition is believed to be due to preferential population of the structure rhomb; this is in agreement with the published assignment of the high energy transition, based on thorough spectroscopic and computational studies. Consequences of a perturbation of the "gas phase" structures axial and rhomb by the protein and solvation are also discussed on the basis of published, experimentally observed structures and spectroscopic data. PMID- 15134936 TI - XSophe-Sophe-XeprView. A computer simulation software suite (v. 1.1.3) for the analysis of continuous wave EPR spectra. AB - The XSophe-Sophe-XeprView computer simulation software suite enables scientists to easily determine spin Hamiltonian parameters from isotropic, randomly oriented and single crystal continuous wave electron paramagnetic resonance (CW EPR) spectra from radicals and isolated paramagnetic metal ion centers or clusters found in metalloproteins, chemical systems and materials science. XSophe provides an X-windows graphical user interface to the Sophe programme and allows: creation of multiple input files, local and remote execution of Sophe, the display of sophelog (output from Sophe) and input parameters/files. Sophe is a sophisticated computer simulation software programme employing a number of innovative technologies including; the Sydney OPera HousE (SOPHE) partition and interpolation schemes, a field segmentation algorithm, the mosaic misorientation linewidth model, parallelization and spectral optimisation. In conjunction with the SOPHE partition scheme and the field segmentation algorithm, the SOPHE interpolation scheme and the mosaic misorientation linewidth model greatly increase the speed of simulations for most spin systems. Employing brute force matrix diagonalization in the simulation of an EPR spectrum from a high spin Cr(III) complex with the spin Hamiltonian parameters g(e) = 2.00, D=0.10 cm(-1), E/D = 0.25, A(x) = 120.0, A(y) = 120.0, A(z) = 240.0 x 10 (-4) cm(-1) requires a SOPHE grid size of N = 400 (to produce a good signal to noise ratio) and takes 229.47 s. In contrast the use of either the SOPHE interpolation scheme or the mosaic misorientation linewidth model requires a SOPHE grid size of only N = 18 and takes 44.08 and 0.79 s, respectively. Results from Sophe are transferred via the Common Object Request Broker Architecture (CORBA) to XSophe and subsequently to XeprView where the simulated CW EPR spectra (1D and 2D) can be compared to the experimental spectra. Energy level diagrams, transition roadmaps and transition surfaces aid the interpretation of complicated randomly oriented CW EPR spectra and can be viewed with a web browser and an OpenInventor scene graph viewer. PMID- 15134938 TI - Effects of polymerization initiator complexation in methacrylated beta cyclodextrin formulations. AB - OBJECTIVES: Methacrylated beta-cyclodextrin (MCD) is a candidate dental monomer that can complex molecules within its hydrophobic cavity. This study determined the effects of complexation of polymerization initiators such as camphorquinone (CQ) and ethyl-4-dimethylaminobenzoate (4E) with MCD on the flexural strength (FS) and degree of conversion (DC) of resulting dental composite formulations. METHODS: Complexation of CQ and 4E with MCD was studied by thin layer chromatography. A mass fraction of 44% 2-hydroxyethylmethacrylate or triethyleneglycoldimethacrylate was mixed separately with a mass fraction of 56% MCD to produce a workable formulation. The mixture was activated with varied amounts of CQ and 4E. One part by mass of the activated resin formulation was mixed with three parts by mass of glass filler. Specimens for FS were prepared by filling molds with composites and curing for 2 min. The cured specimens were immersed in 37 degrees C water for 24 h and FS was measured with an Instron machine at a crosshead speed of 0.5 mm/min. DC in MCD-based resin formulations was measured with a differential photocalorimeter under nitrogen. RESULTS: MCD appears to form inclusion complexes with CQ and 4E. As a result, FS and DC of MCD based composites vary significantly as a function of the concentration of polymerization initiators used in the formulations. SIGNIFICANCE: Complexation of polymerization initiators with MCD can influence the FS and DC in MCD-based dental formulations and should be taken into consideration when evaluating MCD as a dental monomer. PMID- 15134939 TI - Tensile strength and microhardness of treated human dentin. AB - OBJECTIVES: To determine the ultimate tensile strength and Knoop hardness of mineralized, EDTA-treated, sodium hypochlorite (NaOCl)-treated, EDTA-treated resin-infiltrated, and NaOCl-treated resin-infiltrated dentin. METHODS: Dumbell shaped specimens with a cross-sectional area of 0.5 mm2 were prepared from the crowns of extracted human third molars. Specimens were randomly assigned to the following experimental groups: (1) mineralized dentin; (2) 0.5 M EDTA demineralized dentin, pH 7/5 days; (3) 5% NaOCl-deproteinized dentin/2 days; (4) EDTA-treated, Single Bond resin-infiltrated dentin; (5) NaOCl-treated, Single Bond resin-infiltrated dentin. All specimens were tested in tension in a Vitrodyne testing machine at 0.6 mm/min. Knoop microhardness was measured on the fractured edges of specimens in groups 1, 3, 4, and 5. Results were analyzed by ANOVA and SNK tests (p < 0.05). RESULTS: Both EDTA and NaOCl treatments caused significant reductions in the tensile strength and microhardness of mineralized dentin (p < 0.05) with the largest reductions observed after NaOCl treatment (p < 0.05) Resin infiltration of treated dentin resulted in moderate increase of its tensile strength and microhardness, however, the original mineralized values were not recovered (p < 0.05). SIGNIFICANCE: Whenever dentin surfaces are treated with EDTA or NaOCl prior to a clinical bonding procedure, clinicians must be aware that a weak layer may be present at the interface, which may lead to premature failures of resin/dentin bonds. PMID- 15134940 TI - Color and opacity variations in three different resin-based composite products after water aging. AB - OBJECTIVES: Correct shade selection and clinical procedures may be invalidated as a result of intrinsic color shifting with time. The aim of this study was to test the influence of exposure to water on the color stability of three structurally different resin-based composites. METHODS: Six comparable Vita shades of three different resin-based composite systems were selected. For each product at each shade, three disks were made, 1 mm in thickness and 15 mm in diameter. The samples were studied with a spectrophotometer equipped with an integrating sphere. For color determinations, a 50% gray card was used as background, and the data were recorded in the CIE L*a*b* system. Opacity was calculated as the ratio of the reflectance of a specimen disk when backed by a black standard that when backed by a white standard. After the initial measurements, the samples were stored for 30 days in a 60 degrees C water bath and then measured again under the same conditions. The color shifts were calculated using the formula: delta E = [(L1 - L2)2 + (a1 - a2)2 + (b1 - b2)2]1/2. RESULTS: If the standard of delta E < 3.3 as clinically acceptable was taken into consideration, one of the three materials involved in the study showed an unacceptable color shift for all shades. Only one of the three materials remained completely within an acceptable shade shift (delta E < 3.3) after aging. The difference in the color stability between products was significant (p < 0.001). The differences may be due to the different nature of the matrix and could also be correlated with filler particles dimensions. With reference to opacity; the results showed a similar pattern as that of the color and the statistical analysis also correlated. SIGNIFICANCE: From this work, it can be concluded that for some products, aging of composites in water leads to color instability and opacity variation. PMID- 15134941 TI - Tooth deformation patterns in molars after composite restoration. AB - OBJECTIVE: Residual stresses from polymerization shrinkage in composite restorations deform a tooth. This may cause debonding, enamel crack propagation, and post-operative sensitivity. Deformation due to shrinkage has been measured previously at a few discrete points. The purpose of this study was to analyze cuspal deformation pattern of the occlusal portion of molars for various cavity types and sizes after restoration with a light-initiated composite. METHODS: Five extracted human molars were successively prepared as Class I, Class II OM, large Class II OM, and large Class II MOD. The cavities were filled with a light-curing composite using a dentin adhesive system. The occlusal portion of the unrestored cavity and the restoration were digitized with a profilometer. The digitized data of the unrestored and restored tooth were used to calculate the cuspal contour change with Cumulus software. Deformation was visualized as a color contour map. RESULTS: Cuspal deformation showed up in the contour map as a reduction of buccal and lingual contour perpendicular to the surface. Large Class II MODs exhibited the highest cuspal deformation, followed by large OM restorations. Cuspal deformations in Class I and small Class II OM restorations were not significantly different. SIGNIFICANCE: When a composite restoration was cured, the surrounding tooth deformed due to polymerization shrinkage. Cavity type and size affected how much cusps moved inward as a result of polymerization shrinkage. This study quantified and visualized the pattern of cuspal deformation. PMID- 15134942 TI - Distribution of transient properties during polymerization of a light-initiated restorative composite. AB - OBJECTIVES: To assess residual shrinkage stress, transient properties must be defined that describe the composite transformation during polymerization. The purpose of this study was to determine the development and distribution of properties that affect the creation of residual stresses in a light-initiated restorative composite. METHODS: Microhardness and shrinkage strain were experimentally measured during and/or after light-initiated polymerization. The data was acquired for different combinations of light intensities and light exposure times. Light attenuation experiments were used to derive local light intensities inside the composite samples. RESULTS: For the microhardness, a nonlinear correlation was found with the administered light energy, defined as the product of light intensity and exposure time. However, shrinkage strain depended on the initiation intensity rather than the light energy. Higher initiation intensities resulted in higher shrinkage strain rates and values. Microhardness and strain values continued to increase after the light initiation. SIGNIFICANCE: Similar microhardness values, and hence degree of cure and mechanical properties, can be achieved by application of comparable light energy. Therefore, microhardness as a function of light energy can be used to describe transient elastic properties during polymerization. Shrinkage strain, and therefore post-gel shrinkage and residual stress, depends primarily on initiation light intensity. Although mechanical properties achieved at a certain light energy level may be similar, residual stresses may differ depending on initiation intensity. PMID- 15134943 TI - Residual shrinkage stress distributions in molars after composite restoration. AB - OBJECTIVE: Experimental measurements on various restoration configurations have shown that restored teeth deform under the influence of polymerization shrinkage, but actual residual stresses could not be determined. The purpose of this study was to calculate and validate shrinkage stresses associated with the reported tooth deformations. METHODS: Three different restoration configurations were applied in a finite element model of a molar. The composite properties were based on experimentally determined composite behavior during polymerization. The occlusal deformation pattern and the residual stress states of the tooth, restoration, and tooth-restoration interface were calculated using a polymerization model based on the post-gel shrinkage concept. Reported strain gauge measurements and occlusal deformation patterns were used for validation. RESULTS: The shrinkage stresses depended on the configuration and size of the restorations. The tooth's resistance against polymerization shrinkage diminished with loss of dental hard tissue. Larger restorations resulted in lower stress levels in the restoration and tooth-restoration interface, but increased stresses in the tooth. The maximum stress values found for different configurations were not decisively different. SIGNIFICANCE: The validated model indicated that shrinkage stress cannot be based on composite properties or restoration configuration alone, but has to be approached as a distributed pattern that depends on the location and on the properties of tooth and restoration, geometry, constraints, and restoration procedures. Tooth deformation was indicative of stresses in the tooth rather than in the restoration or across the tooth restoration interface. PMID- 15134944 TI - Influence of mechanical and chemical polishing in the solubility of acrylic resins polymerized by microwave irradiation and conventional water bath. AB - OBJECTIVES: The aim of this work was to evaluate the solubility of acrylic resin activated by microwave irradiation (MI) or water bath (WB), when submitted to chemical (CP) or mechanical (MP) polishing. METHODS: Forty acrylic resin samples were made and processed either by water bath (74 +/- 1 degrees C, 9 h) or microwave irradiation (500 W, 3 min). After deflasking, the samples were finished with aluminum oxide sandpapers in decreasing granulations till reaching similar dimensions. The samples were divided into four groups according to the association between kind of polymerization and polishing: A (WB + CP), B (WB + MP), C (MI + CP) and D (MI + MP). Solubility test was performed for each group and percentile solubility was calculated. Data were statistically analyzed using variance analysis and Kruskal-Wallis. RESULTS: The average of percentile solubility (%) was obtained: A = 0.07, B = 0.02, C = 0.04, D = -0.14, however, no significant difference was found between types of polishing in the samples polymerized by water bath (A and B). When processed by microwave irradiation (C and D), there was significant difference between the applied methods of polishing, so that mechanical polishing lead to a lower solubility. SIGNIFICANCE: Solubility is a property of acrylic resins, representing not reacted substances releasing that could promote tissular reactions in prosthesis users. The association between polymerization by microwave irradiation and mechanical polishing showed less residual substances releasing for heat-cured acrylic resins, reducing the probability of developing tissular reactions. PMID- 15134945 TI - Organosilicon dental composite restoratives based on 1,3-bis[(p-acryloxymethyl) phenethyl] tetramethyldisiloxane. AB - OBJECTIVES: The major concern associated with the use of polysiloxanes as polymer matrices in dental restorative materials, is the generally modest mechanical properties of the polymers. However, it has long been demonstrated that thermal stability, and mechanical properties of polysiloxanes can be substantially modified by incorporation of bulkier substituents such as phenyl groups or more polar groups in the chains. The purpose of this research was to evaluate visible light activated dental composites based on the high molecular weight siloxane monomer 1,3-bis[(p-acryloxymethyl) phenethyl] tetramethyldisiloxane (BAPD). METHODS: Hardness, diametral tensile strength (DTS), degree of conversion (DC), water sorption (WS) and polymerization shrinkage of BAPD-based composites and bis GMA-based composites were determined and compared. RESULTS: Composites based on BAPD exhibited low WS, high DC, low polymerization shrinkage, and had hardness and DTS values that were not significantly lower than those of dental composites based on bis-GMA. SIGNIFICANCE: BAPD is a high molecular weight monomer (MW = 511) with a low viscosity. It did not require the use of low molecular weight diluent monomers in formulating composite resins. The DC of BAPD was high, ranging from 86 to 94%. Although the DC of BAPD was significantly higher than the conventional difunctional dental monomers, the polymerization shrinkage of the siloxane composites (1.70 - 1.81 vol%) was comparable to several composites based on bis-GMA. PMID- 15134946 TI - Influence of c-factor and layering technique on microtensile bond strength to dentin. AB - OBJECTIVE: Due to polymerisation shrinkage of resin-based composites, a high configuration factor in deep Class I cavities leads to a certain amount of stress when the material is bonded. The aim of this in vitro study was to evaluate the influence of c-factor and different layering approaches on bonding to dentin with three different adhesive systems. MATERIALS AND METHODS: Dentin bond strengths of Z250 bonded with OptiBond FL, Single Bond, and One Up Bond F were measured on flattened dentin surfaces without cavity walls and on the cavity floor of Class I cavities (10 layering concepts). The resin composite increments were applied horizontally, vertically and obliquely, both with and without a flowable liner. The tests were carried out in a microtensile apparatus at a crosshead speed of 1 mm/min after 24 h of storage at 37 degrees C in water. Mean bond strengths were analysed using the Wilcoxon test and multiple comparisons according to the Mann Whitney U-test. Specimens having failed prior to the bond strength test were included as 0 MPa. RESULTS: The groups bonded on flat surfaces exhibited significantly higher bond strengths than specimens cut from filled cavities. Within the cavity groups, OptiBond FL and Single Bond exhibited no significant differences, however, being above One Up Bond F. Within the groups of each adhesive, major differences between the layering concepts were detectable. Bulk technique led to low dentin adhesion at the cavity floor, above all for Single Bond and One Up Bond F. Horizontal layers resulted in significantly higher bond strengths than did vertical or oblique. Lining with a flowable composite did not promote bond strength for OptiBond FL. For the other adhesives, a lining improved adhesion when vertical or oblique layers were applied, for horizontal increments no effect was evident. CONCLUSIONS: The c-factor is an influencing factor for dentin adhesion. However, using an appropriate layering technique, high bond strengths to deep cavity floors can be achieved. PMID- 15134947 TI - Effect of drying time of 3-methacryloxypropyltrimethoxysilane on the shear bond strength of a composite resin to silica-coated base/noble alloys. AB - OBJECTIVES: In this in vitro study, the effect of various drying (surface reaction) times of a commercial silane, other than that recommended by the manufacturer (at least 5 min), on the bond strength between the resin composite and silica coated base and noble alloys was evaluated. METHODS: A total of 112 disc specimens (9 mm diameter and 0.5 mm thickness) were cast out of two types of alloy designed for ceramic firing, one of which was a noble (Degunorm) (gold silver-platinum) and the other a base alloy (Wiron 99) (nickel-chromium molybdenum). The specimens were assigned to two main groups according to each alloy type. These two main groups were further divided into seven subgroups, having eight specimens each. The specimens of both alloy types were air-abraded with 30 microm silica (SiO2) coated alumina (Al2O3) (CoJet-Sand, ESPE, Seefeld, Germany). The conditioned surfaces were coated with 3 methacryloxypropyltrimethoxysilane (MPS) and were allowed to react and dry for 1, 2, 3, 4, 5, 6, and 7 min, respectively, before the opaquer was applied. Immediately after the waiting periods for the silane to dry, first opaquer and then resin composite were applied. After storage in water for 30 days at 37 degrees C and thermocycling (5000 cycles, 5-55 degrees C), shear tests were performed using the universal testing machine at a crosshead speed of 0.5 mm/min. RESULTS: Analysis of data showed no significant difference in bond strength for any silane drying and reaction period for both base and noble alloys between 1 and 7 min (ANOVA, P = 0.05) (Degunorm: 5.8 - 7.4 MPa and Wiron 99: 7.2 - 10.2 MPa, respectively). Bond strengths of resin composite to base alloys were significantly higher than those to noble alloys at 2, 3 and 5 min (P = 0.0045, P = 0.05, P = 0.002, respectively). SIGNIFICANCE: In order to optimize the flow of laboratory work, the silane solution drying time might be reduced to 1 min for both base and noble alloys. PMID- 15134948 TI - Evaluation of fracture toughness of a fiber containing dental composite after flexural fatigue. AB - OBJECTIVE: The purpose of this study was to examine the effect of cyclic loading in four different environments on the fracture toughness of a fiber reinforced resin composite. METHODS: The specimens were 3 x 3 x 25 mm3 bars polished with 320 grit SiC paper. A 60 degrees v-notch was machined 1 mm deep at the midspan of each bar. The specimens were loaded in air, artificial saliva, water, and a 50/50 by volume mixture of ethanol and water at a frequency of 5 Hz with sinusoidal loads cycling between 10 and 20 N for 1, 1000, 10,000 and 100,000 cycles. Specimens were also aged for 4 months in each respective media. Following cyclic loading, the specimens were tested in three-point loading. Statistical analysis consisted of four 2-way ANOVAs followed by Tukey's HSD inference, and two 3-way ANOVAs followed by two-sample t-tests. RESULTS: The ANOVAs indicated no difference in the means of fracture toughness for the set numbers of cycles, but a significant difference for the aging solutions and the aging times. SIGNIFICANCE: Statistical analysis showed a significant decrease of fracture toughness from unaged to aged specimens when aged in air, saliva, and 50/50 ethanol/water. The latter showed the most pronounced decrease. The effect of the 50/50 by volume mixture of ethanol and water would appear to attack the resin matrix and/or the bond between the resin matrix and the filler, which resulted in the observed decrease in fracture toughness. In addition, the significant decrease in fracture toughness following aging is attributed to residual matrix stress around the large fiber filler particles resulting in separation of the fiber filler from the resin matrix. PMID- 15134949 TI - Compressive creep and recovery of light-cured packable composite resins. AB - OBJECTIVE: The purpose of this investigation was to study the time-dependent creep and recovery behavior of some packable composites resins compared to a typical dispersion-phase amalgam alloy. METHODS: Three packable posterior composite resins (ALERT, SureFil, Solitaire) were used as experimental groups and a high copper admixed amalgam alloy (Dispersalloy) was chosen as a control material. Cylindrical specimens (4 mm x 8 mm) were fabricated and stored in artificial saliva for 1 week at 37 degrees C. A custom-made creep testing machine was used to test the compressive creep of each of the materials under specified conditions of stress and temperature (155.5 MPa and 37 degrees C, 155.5 MPa and 55 degrees C; 36 MPa and 37 degrees C; 36 MPa and 55 degrees C) for 24 h, after which the strain recovery was recorded over a period of 24 h. RESULTS: One-way ANOVA and Fisher's test showed no significant differences in creep and recovery strain behaviors between ALERT and SureFil for each test condition (P > 0.05). However, for residual strain, there was a highly significant difference between Dispersalloy and each of the packable composites (P < 0.01), but no significant differences between the composites themselves (P > 0.05). ALERT and SureFil had the lowest creep strain (1.91 +/- 0.59% and 1.96 +/- 0.36%, respectively) while Solitaire showed 6.09 +/- 0.26% creep strain at 155.5 MPa and 37 degrees C. SIGNIFICANCE: The low creep and residual strain values of ALERT and SureFil compared to Dispersalloy amalgam suggest that these packable composite resins are suitable for restoring stress-bearing areas because of their ability to withstand viscoelastic deformation better than amalgam, over extended time periods, even under high load and temperature. PMID- 15134950 TI - Rapid analysis of metallic dental restorations using X-ray scanning analytical microscopy. AB - OBJECTIVES: X-ray scanning analytical microscopy (XSAM) makes it possible to analyze small specimens in air without pretreatment. The purpose of this study was to utilize XSAM for the rapid analysis of metallic dental restorations by microsampling. METHODS: Six different dental alloys were scratched with brand-new silicone points to obtain metal on the silicone point for compositional analysis. The fluorescent spectra of XSAM were measured to determine the metal attached to the specimen. RESULTS: The major components of the six dental metals, except for palladium, were clearly detected. The identification of palladium was difficult since the fluorescent X-ray of palladium is quite close to that of rhodium, which is the source metal of the incident X-rays. However, with a slight modification of XSAM, palladium was also identified. The total time required for sampling and analysis with XSAM was less than 10 min. The amount of the attached metal was estimated to be less than 30 microg. This amount of sampling does not damage metal restorations. SIGNIFICANCE: XSAM analysis using the microsampling technique is useful for the rapid analysis of metallic restorations. PMID- 15134951 TI - Urethral incompetence: an update with special reference to residual maximal urethral closure pressure. PMID- 15134952 TI - Applying complexed prostate-specific antigen to clinical practice. PMID- 15134953 TI - Robot-assisted versus open radical prostatectomy: a comparison of one surgeon's outcomes. AB - OBJECTIVES: To compare internally one surgeon's standard open radical prostatectomy (RP) and robot-assisted laparoscopic RP (RLP) results. RLP, like standard laparoscopic RP, ultimately needs to produce similar or improved results compared with standard RP techniques. Little information comparing RLP with standard RP exists. METHODS: As an internal control, we selected the last 60 standard RPs performed by one surgeon (T.A.) before initiating RLPs. For the RLP group, we selected cases 46 to 105 (n = 60) after the learning curve had adequately matured. We compared the clinical characteristics, perioperative results, and early clinical outcomes. RESULTS: The study and control groups had similar clinical characteristics (age, body size, preoperative prostate-specific antigen level, clinical stage, and Gleason score). No statistically significant differences were found between groups for prostate size, pT stage, Gleason score, or margin status (16.7% versus 20%; P = nonsignificant). The RLP group had a statistically significant advantage for estimated blood loss (103 versus 418 mL), postoperative hemoglobin change (1.6 versus 3.3 mg/dL), and hospital stay (1.02 versus 2.2 days). Complete continence (0 pads) at 3 months of follow-up and the rate of postoperative complications were similar for the RLP and RP groups (76% versus 75% and 6.7% versus 10%, respectively). CONCLUSIONS: We present the results of RLP and RP performed by one surgeon. With only a 100-case experience, RLP had oncologic and urinary outcomes that were at least equal to those after RP. RLP offers the benefits of minimally invasive surgery and does not compromise clinical or pathologic outcomes. PMID- 15134954 TI - Distinguishing stent from stone: use of bone windows. AB - OBJECTIVES: To present a simple method to assist in distinguishing stents or nephrostomy tubes from urinary tract stones. Non-contrast computed tomography (CT) is now routinely used for the diagnosis and follow-up of urinary tract stones. When stents or nephrostomy tubes are in place, it may be difficult to differentiate the stent from the stone. METHODS: Three representative patients with ureteral stents or nephrostomy drainage catheters (stents/tubes) who were suspected of having residual urinary tract stones were studied by visualizing the suspicious density on CT. Abdominal windows were used for initial viewing, then bone windows were used, and the two techniques were compared. The Hounsfield units (HU) of the various stents/tubes were measured in vitro to establish reference points. The pixel densities of stones have a known range and, in our experience, may be as high as 1600 HU. RESULTS: Using the abdominal window, no difference was observed in the radiographic appearance of the stents/tubes compared with urinary calculi. However, when viewed in the bone window, the stone appeared less dense than the stent/tube. The in vitro pixel densities of the stents/tubes measured 1600 to 2600 HU compared with the calculi, which do not exceed 1600 HU. CONCLUSIONS: Using the abdominal window, stents and stones may have the same CT appearance. However, the bone window allows a visual distinction between a stent/tube and a stone. This distinction is accounted for by differences in pixel density. This observation allows one to distinguish a stent/tube from a stone on CT in patients in whom a nephrostomy tube or ureteral stent is present. PMID- 15134957 TI - Helical computed tomography accurately reports urinary stone composition using attenuation values: in vitro verification using high-resolution micro-computed tomography calibrated to fourier transform infrared microspectroscopy. AB - OBJECTIVES: To assess the ability of helical computed tomography (CT) to differentiate regions of known mineral composition in typical, heterogeneous urinary stones. Interest is substantial in the urologic community in using radiologic imaging to determine accurately the composition of urinary calculi. Recent advances in CT make this a viable prospect, but the heterogeneity of most stones is a complicating factor. METHODS: The ability of micro-CT (a high resolution laboratory instrument) to identify the mineral composition of stones was confirmed by calibrating micro-CT attenuation values to pure mineral regions of sliced stones using infrared microspectroscopy. Intact human urinary stones were then analyzed by micro-CT, and regions-of-interest of pure mineral were correlated with identical regions-of-interest from quad slice multi-detector row helical CT images. With helical CT, narrow slice widths were used to decrease volume-averaging errors, and bone windows were used so that internal stone structure was visible. RESULTS: When stones were imaged using helical CT at narrow slice widths, mineral-specific regions-of-interest yielded nonoverlapping attenuation values for uric acid (566 to 632 Hounsfield units [HU]), struvite (862 to 944 HU), calcium oxalate (1416 to 1938 HU), and hydroxyapatite (2150 to 2461 HU). CONCLUSIONS: High resolution helical CT yields unique attenuation values for common types of stone mineral, but proper windowing is required to localize regions of homogeneity. The results of this in vitro study suggest that high-resolution helical CT may be able to identify stone composition at patient diagnosis. PMID- 15134958 TI - Systematic evaluation of ureteral access sheaths. AB - OBJECTIVES: To compare the physical characteristics of ureteral access sheaths that affect their ability to resist buckling and kinking in the ureter. METHODS: Eight commercially available ureteral sheaths were tested. The buckling pressure was measured by adding sequential loads to a point 20 cm from the tip of the sheath until the mass that resulted in buckling of the catheter was determined. The kinking pressure was determined by measuring the diameter of the sheath with a digital caliper as sequential loads were applied to the surface of the sheath. The frictional properties of the sheath were measured by testing the force required to push the sheath through a mock tube. RESULTS: The Cook Flexor was more resistant to buckling, requiring 202 g of force before buckling occurred, and both the Cook Flexor and the Applied access sheaths were more lubricious. The kinking measurements demonstrated a linear loss of diameter of the sheath with added loads. The Applied Forte XE and Cook Flexor ureteral access sheaths were more resistant to kinking than were the other sheaths tested. CONCLUSIONS: Understanding the physical characteristics of ureteral sheaths may help determine which sheaths will perform well with regard to buckling during insertion and kinking during use. PMID- 15134959 TI - Anastomotic urethroplasty for failed previously treated membranous urethral rupture. AB - OBJECTIVES: To determine whether delayed excision and primary anastomosis is appropriate after failed previous therapeutic attempts for post-traumatic membranous urethral strictures. Delayed excision and primary anastomosis is widely accepted as the first-line treatment of post-traumatic membranous urethral strictures. METHODS: A review of the medical records identified 13 patients who had undergone anastomotic urethroplasty as a second procedure for traumatic membranous urethral strictures. The previous treatments in these patients included anastomotic urethroplasty in four, staged urethroplasty in four, and endoscopic urethrotomy or primary catheter realignment in five. The mean stricture length was 2.8 cm. All patients underwent excision and primary bulboprostatic anastomosis using the perineal approach. RESULTS: Four patients required partial pubectomy and one required corporal rerouting to achieve anastomosis. The mean follow-up was 27 months. Urethrography performed 1 month postoperatively demonstrated a widely patent anastomosis in all cases. Flexible urethroscopy performed 1 year after surgery revealed a widely patent anastomosis with normal urethral mucosa in all patients but one. The mean maximal flow rate at the last follow-up visit was 23.5 mL/s compared with 3.9 mL/s preoperatively. No statistically significant postvoid residual urine volume was found in any patient. One patient developed an anastomotic stricture 3 months after surgery that was treated successfully by internal urethrotomy. Thus, the objective success rate was 92%. Subjectively, all patients but one reported satisfactory voiding. Complications were mild and included urinary tract infection, bladder stone formation, and decreased erectile function in 1 patient each. CONCLUSIONS: Even in patients with failed previous surgical attempts, excision and primary anastomosis is feasible and provides good surgical results in post-traumatic posterior urethral strictures. The complications were mild and easily treated. PMID- 15134960 TI - Use of American Society of Anesthesiologists physical status classification to assess perioperative risk in patients undergoing radical nephrectomy for renal cell carcinoma. AB - OBJECTIVES: To perform a retrospective analysis to determine the operative morbidity in patients with substantial comorbidities requiring renal surgery. Increasing numbers of patients requiring renal surgery are presenting with substantial comorbidities, such as diabetes mellitus, chronic obstructive pulmonary disease, and cardiovascular disease. METHODS: The American Society of Anesthesiologists (ASA) physical status classification was used to define perioperative risk. Of 1087 patients who underwent nephrectomy between 1989 and 2001, 237 patients were classified as ASA classification 1 or 2 (low risk), 297 were ASA classification 3 (intermediate risk), and 17 were ASA classification 4 (high risk). RESULTS: No statistically significant differences were found among the low-risk, intermediate-risk, or high-risk patients with regard to 1997 T stage distribution, mean tumor size, vascular and/or inferior vena cava involvement, percentage of partial nephrectomy, adjacent organ resection, or preoperative hemoglobin. Intermediate-risk patients did have a greater estimated blood loss (946 versus 739 mL, P = 0.05), leading to greater transfusion rates (42% versus 28%, P = 0.001). However, no increase occurred in intraoperative or postoperative morbidity. High-risk patients also had greater transfusion rates, as well as a greater rate of complications occurring more than 24 hours after surgery. CONCLUSIONS: Partial or radical nephrectomy can be offered to patients with comorbid conditions. ASA classification 3 patients are more likely to require transfusion. This may have been a result of a lower threshold to transfuse patients with preoperative morbidities. However, the perioperative and postoperative complication rates were similar to those of low-risk patients. Not surprisingly, high-risk patients had greater rates of transfusions and complications. PMID- 15134963 TI - Comparative study of ureteral stripping versus open ureterectomy for nephroureterectomy in patients with transitional carcinoma of the renal pelvis. AB - OBJECTIVES: To evaluate the clinical outcome of nephroureterectomy with endoscopically assisted transurethral ureteral stripping for transitional cell carcinoma of the renal pelvis in a comparative study. METHODS: Sixty patients with localized renal pelvic cancer were enrolled in a prospective comparative nonrandomized study. Of these, 28 patients underwent nephroureterectomy with endoscopically assisted transurethral ureteral stripping and 32 underwent conventional nephroureterectomy with a bladder cuff. Both short-term and long term results were analyzed in this series. RESULTS: The operating time for patients with ureteral stripping was significantly shorter than for those with a standard two-incision nephroureterectomy (median 183 versus 250 minutes, P = 0.0231), and the amount of blood loss was significantly less (median 150 versus 390 mL, P = 0.0002). Intravesical recurrence was detected in 10 (35.7%) of the 28 patients with ureteral stripping, and the 1-year and 3-year recurrence-free rate was 68.0% and 57.7%, respectively. Seven patients treated by the standard two incision nephroureterectomy (21.9%) experienced intravesical recurrence, with a 1 year and 3-year recurrence-free rate of 96.8% and 75.0%, respectively. The recurrence rate was significantly greater in the group with ureteral stripping (P = 0.0287). CONCLUSIONS: Compared with conventional nephroureterectomy with a bladder cuff, nephroureterectomy with transurethral stripping is a minimally invasive procedure with a shorter operating time and less blood loss, but a statistically significantly greater intravesical recurrence rate. Greater consideration should be taken before selecting this procedure. PMID- 15134964 TI - Laparoscopic-assisted renal autotransplantation. AB - OBJECTIVES: To report our experience with laparoscopic nephrectomy and autotransplantation for the management of a variety of conditions with significant loss of healthy ureteral tissue or ureteral length. Renal autotransplantation has been described as an effective method for addressing this problem, avoiding the need for nephrectomy or complex ureteral replacement. In an effort to decrease the morbidity associated with traditional autotransplantation we elected to perform laparoscopic procurement of the kidney. METHODS: Four patients underwent laparoscopic nephrectomy using a transperitoneal four-port technique and subsequent autotransplantation into the iliac fossa for the treatment of proximal ureteral avulsion (2 patients), ureteral malignancy, and ureteral stricture. All patients had less than 5 cm of viable ureter. RESULTS: All procedures were performed without intraoperative complications. All renal scans on postoperative day 1 demonstrated good perfusion. None of the patients had a postoperative rise in serum creatinine. On postoperative day 1, the mean creatinine value was 0.95 mg/dL. Three patients had an uneventful postoperative course. One patient, however, had loss of the graft because of renal vein thrombosis on postoperative day 7. She was later found to have an undiagnosed thrombophilic disorder (decreased levels of antithrombin III) and to have a recent history of oral contraceptive use. CONCLUSIONS: Laparoscopic nephrectomy with renal autotransplantation is a feasible minimally invasive alternative to treat patients who have significant ureteral loss. This approach avoids the need for an upper abdominal or flank incision, resulting in decreased morbidity. The initial follow-up studies indicated stable renal function. Additional long-term observation is currently under way. PMID- 15134965 TI - Right retroperitoneal versus left transperitoneal laparoscopic live donor nephrectomy. AB - OBJECTIVES: To describe our preferred method of right laparoscopic live donor nephrectomy (LDN) using a retroperitoneoscopic approach to determine the indications for, and overall rate of, right LDN and to compare the donor and recipient early outcomes of right retroperitoneal LDN to those of left transperitoneal LDN in a consecutive single-institution series. METHODS: At our institution, LDN for allotransplantation was performed in 143 consecutive patients. The indications for right LDN (n = 29) included multiple left renal vessels (n = 18), early branching of the left renal artery (n = 1), left renal vein anomaly (n = 2), right renal arterial fibromuscular dysplasia (n = 2), right renal cyst (n = 3), mild right hydronephrosis with delay on renal scan (n = 1), or right nephrolithiasis (n = 2). RESULTS: Right LDN was performed in 29 (20.3%) of 143 patients using a retroperitoneal approach in all but the first case. Right retroperitoneal LDN was associated with decreased blood loss and operative time compared with left transperitoneal LDN. The hospital stay, analgesic use, and donor serum creatinine at discharge were similar in both groups. Despite a statistically significantly increased warm ischemia time and decreased renal vein length, right retroperitoneal LDN was associated with recipient functional outcomes at 5 and 30 days after transplant that were no different from those after left transperitoneal LDN. CONCLUSIONS: Right retroperitoneal laparoscopic LDN provides similar donor and recipient outcomes when compared with the left transperitoneal approach and obviates most of the technical challenges encountered with a right transperitoneal approach. PMID- 15134966 TI - Impact of surgical volume on mortality and length of stay after nephrectomy. AB - OBJECTIVES: To evaluate the volume-outcome relationship in patients undergoing nephrectomy for neoplastic disease by examining the impact of the number of cases performed on in-hospital mortality and length of stay. Surgical volume is associated with postoperative mortality for many complex procedures; however, this relationship has not been characterized for patients undergoing nephrectomy for neoplastic disease. METHODS: Using the Nationwide Inpatient Sample database, 20,765 patients who underwent nephrectomy for neoplasm from 1993 through 1997 were identified by International Classification of Disease, Ninth Revision codes. Cases were stratified into volume groups on the basis of annual nephrectomy rates: low-volume hospitals performed 1 to 14 nephrectomies per year, medium volume hospitals performed 15 to 33 per year, and high-volume hospitals performed more than 33 per year. Unadjusted and risk-adjusted analyses were performed. RESULTS: Overall mortality was 1.39%. Mortality declined as surgical volume increased. The mortality rate for low-volume hospitals was 1.60% versus 1.49% for medium-volume hospitals and 1.04% for high-volume hospitals (P = 0.017). After adjusting for case mix, high-volume hospitals had a 32% lower risk of in-hospital mortality than medium-volume hospitals (P = 0.029) and a 25% lower risk than low volume hospitals (P = 0.094). Length of stay was not affected by hospital volume. Other independent risk factors for in-hospital mortality included age older than 65 years, chronic pulmonary disease, metastatic disease, and the urgent nature of the admission. CONCLUSIONS: A greater surgical volume, age younger than 65 years, elective conditions, and less comorbidity are associated with a significantly decreased risk of in-hospital mortality after nephrectomy. These findings provide compelling evidence that hospital volume and patient characteristics have important effects on surgical outcome specific to renal neoplasms. PMID- 15134967 TI - Urodynamic evidence of effectiveness of botulinum A toxin injection in treatment of detrusor overactivity refractory to anticholinergic agents. AB - OBJECTIVES: To investigate the urodynamic changes after detrusor injection of botulinum A toxin in patients with detrusor overactivity refractory to treatment with anticholinergic agents. METHODS: Thirty patients with detrusor overactivity refractory to anticholinergic agents were treated with detrusor injection of botulinum A toxin (Botox) 200 U at 40 sites. Urodynamic parameters and symptom scores were assessed at baseline and 2 weeks and 3 months after the injections. Patients' responses were classified as excellent, improved, or failed. RESULTS: The 12 female and 18 male patients were aged 7 to 83 years (mean 67 +/- 17). Of the 30 patients, 12 had neurogenic detrusor overactivity, 8 had idiopathic detrusor overactivity, and 10 had previous bladder outlet obstruction or had undergone previous transurethral prostatectomy. After detrusor botulinum A toxin injection, 8 patients regained urinary continence (26.7%), 14 patients had improvement in frequency, urgency, and incontinence (46.7%), and treatment failed 8 patients (26.7%). The total success rate was 73.3%. Four patients experienced transient urinary retention and six had difficulty urinating after treatment. The urodynamic results showed increased cystometric capacity, decreased voiding pressure, increased postvoid residual urinary volume, decreased voiding efficiency, and increased bladder neck opening time at 2 weeks. The voiding pressure remained low and bladder neck opening time remained increased at 3 months, although the postvoid residual volume and voiding efficiency had returned to baseline levels. The therapeutic effects lasted for 3 to 9 months (mean 5.3). CONCLUSIONS: Detrusor injection of 200 U of botulinum A toxin is effective in the treatment of detrusor overactivity that is refractory to anticholinergic agents. Patients with detrusor overactivity and inadequate contractility should be carefully selected for this procedure because the postvoid residual urine volume may increase after treatment. PMID- 15134968 TI - Impact of transurethral resection of bladder tumor: analysis of cystectomy specimens to evaluate for residual tumor. AB - OBJECTIVES: To analyze the impact of transurethral resection of bladder tumor (TURBT) on the outcome of patients with bladder cancer who undergo subsequent cystectomy. METHODS: We reviewed the records of 90 patients with pathologic Stage T2 or less transitional cell carcinoma of the bladder at our institute. Before radical cystectomy, TURBT was performed for diagnostic and therapeutic purposes in all patients. We used the term "complete TURBT" in the following situations: no residual tumor endoscopically after TURBT, the presence of a muscle layer in the TURBT specimen, and no gross residual tumor in the cystectomy specimen as evaluated by a pathologist. RESULTS: No statistically significant differences in the completeness of TURBT were observed by age, sex, tumor multiplicity, shape, history of previous TURBT, tumor grade, or stage. Of 21 patients with superficial bladder tumors who had undergone incomplete resection, 14 (66.7%) had a higher stage on the cystectomy specimen; 3 (17.7%) of 17 patients had a higher stage in the complete TURBT group. Of 34 patients with an invasive tumor who underwent complete TURBT, 10 (29.4%) had an identical pathologic stage after cystectomy; the remaining patients had a lower stage. However, only 3 (16.7%) of 18 patients had a lower stage in the incomplete resection group. Of patients with Stage T2, those with pT0 stage had a better chance of survival than those with residual tumor (P = 0.04). CONCLUSIONS: Our findings indicate that the completeness of TURBT is an important aspect of patient outcome in cases of superficial or invasive bladder cancer. PMID- 15134970 TI - Effect of intravesical instillation on performance of uCYT+ test. AB - OBJECTIVES: To assess the influence of intravesical instillation of epirubicin or bacille Calmette-Guerin (BCG) in patients with bladder cancer on the clinical performance of the uCyt+ test, which traces the three monoclonal antibodies M344, LDQ 10, and 19A211 in urothelial cells. METHODS: A total of 267 patients with a mean age of 70.5 years (range 35 to 87) were included in this prospective study. All patients were being followed up after complete transurethral resection of superficial urothelial cancer. All patients received a single instillation of 80 mg epirubicin within 24 hours of transurethral resection and 163 underwent subsequent cycles of chemotherapy with epirubicin or immunotherapy with BCG. This was because of multifocality, high-recurrence, or high-grade urothelial cancer. At the next control, all patients underwent urinary cytology, with the uCyt+ test performed on liquid-based cytology (ThinPrep), subsequent cystoscopy, and evaluation of any suspicious lesion by biopsy. Patients who underwent intravesical BCG therapy underwent random bladder cold biopsy after every instillation series. RESULTS: Of the 267 patients, 8 could not be evaluated by uCyt+ because of insufficient cells. Of the remaining 259 patients under assessment, 86 had proven urothelial cancer. In the 101 patients under assessment with only a single intravesical instillation of epirubicin (group 1), the overall sensitivity for cytology and the uCyt+ test was 44.7% and 76.6%, respectively. Cytology had a specificity of 90.7% and uCyt+ of 64.8%. The negative predictive value for cytology and uCyt+ was 65.3% and 76.1%, respectively. Of the 158 patients under evaluation who were treated with long-term intravesical therapy (group 2), the overall sensitivity and specificity for cytology and the uCyt+ test was 37.8% and 84.6% and 94.9% and 70.6%, respectively. The negative predictive value for cytology and the uCyt+ test was 83.1% and 93.3%, respectively. Statistical analysis showed no statistically significant difference between the overall sensitivity (P = 0.512) and specificity (P = 0.558) for the uCyt+ test in the two groups of patients. CONCLUSIONS: The sensitivity and specificity of the uCyt+ test are not significantly affected by intravesical long term treatment with epirubicin or BCG. PMID- 15134971 TI - Cost aspects of transurethral resection of the prostate, contact laser prostatectomy, and electrovaporization. AB - OBJECTIVES: To compare the costs of transurethral resection of the prostate (TURP), contact laser prostatectomy (CLP), and electrovaporization in men with lower urinary tract symptoms associated with benign prostatic hyperplasia. METHODS: We conducted a randomized controlled trial that included men with lower urinary tract symptoms who met the criteria of the International Scientific Committee on benign prostatic hyperplasia. Subjective changes were quantified using questionnaires validated by the American Urological Association. The maximal free urinary flow rate was estimated. Morbidity and mortality were registered. These parameters were measured at regular intervals for up to 1 year and once during long-term follow-up. A cost analysis together with a sensitivity analysis was performed on the basis of a follow-up of 12 months. RESULTS: A total of 50 men were randomized to TURP, 45 to laser treatment, and 46 to electrovaporization. The subjective and objective changes were very similar during the 12 months of follow-up. The costs were highest for CLP (1885 dollars), followed by TURP (1707 dollars), and were lowest for electrovaporization (1489 dollars). However, the length of hospital stay decreased during the trial more for CLP and electrovaporization than for TURP. Recalculations demonstrated almost equal costs for CLP and TURP (1697 dollars and 1643 dollars, respectively) and the lowest costs for electrovaporization (1386 dollars). CONCLUSIONS: Electrovaporization has a better cost-effectiveness than CLP and TURP in patients with lower urinary tract symptoms associated with benign prostatic hyperplasia. CLP and TURP showed very similar cost-effectiveness ratios. PMID- 15134972 TI - Biopsy indication--a predictor of pathologic stage among men with preoperative serum PSA levels of 4.0 ng/mL or less and T1c disease. AB - OBJECTIVES: To determine the indications for biopsy among men with a prostate specific antigen (PSA) level of 4 ng/mL or less and T1c disease and whether the reason for biopsy correlated with the final pathologic stage. It is not uncommon for men to undergo radical prostatectomy (RP) with a prostate-specific antigen (PSA) level of 4 ng/mL or less and T1c disease. METHODS: We examined the percentage of men with T1c disease and PSA level of 4.0 ng/mL or less undergoing RP over time. We identified 874 men with T1c prostate adenocarcinoma and PSA levels of 4 ng/mL or less who were treated with RP between 1994 and 2003. We excluded 72 patients who had received preoperative hormonal therapy and 86 patients for whom the reason for biopsy was unclear. The pathologic outcomes were compared between the various biopsy indication groups using analysis of variance. RESULTS: The percentage of men with T1c disease and a PSA level of 4.0 ng/mL or less has steadily increased from 0% before 1990 to 19% in 2003. Among men treated between 1994 and 2003, the most common indication for biopsy was a spike in the PSA level to greater than their reference level before biopsy (n = 471, 66%), although in all patients, the PSA level returned to 4 ng/mL or less before surgery. In 163 patients (35%), the PSA level remained elevated above the reference range and in the remaining 308 patients (65%), the PSA declined to a value below the reference range before surgery. The second most common indication for biopsy was an abnormal digital rectal examination by the referring/primary physician that, when repeated by the attending surgeon, was believed to be normal (n = 155, 22%). Abnormal PSA velocity accounted for biopsy in 68 patients (10%). Among patients with the three most common indications for biopsy, no statistically significant differences in the pathologic tumor characteristics between the groups were noted on the basis of the biopsy indication. CONCLUSIONS: One in five men undergoing RP at our institution today has T1c disease and a PSA value of 4.0 ng/mL or less. The most common indication for biopsy was an elevated PSA level above their reference range, although in most, the PSA had returned to a normal value before surgery. Regardless of the indication for biopsy, these men had favorable findings at RP. That 22% of patients underwent biopsy because of what was perceived to be an abnormal digital rectal examination underscores the need to continue to perform digital rectal examination along with PSA determination as a part of routine prostate cancer screening. PMID- 15134973 TI - No reason for immediate repeat sextant biopsy after negative initial sextant biopsy in men with PSA level of 4.0 ng/mL or greater (ERSPC, Rotterdam). AB - OBJECTIVES: In the early detection of prostate cancer (CaP) uncertainty exists concerning the most appropriate biopsy procedure. Within the European Randomized Study of Screening for Prostate Cancer (ERSPC) lateralized sextant biopsies are used. False-negative results of sextant biopsies have led to the extensive use of procedures using 12 or more biopsy cores. The ERSPC offers the opportunity to study the yield of repeat biopsies after 4 years in men who had negative sextant biopsies and a prostate-specific antigen (PSA) level of 4.0 mg/mL or more at the first screening round. METHODS: Between August 1996 and May 1998, a total of 6876 men (age 55 to 74 years) were randomized to the screening arm and actually underwent screening. The numbers and levels of biopsy indicators, as well as possible predictors for biopsy outcome, in the second screening round, such as prostate volume, volume change over time, prostate-specific antigen density (PSAD), PSA velocity, and age, were calculated and compared for participants with positive and negative biopsies in round 2. The positive predictive value (PPV) and detection rates, as well as parameters of aggressiveness, were evaluated for second-round biopsy-detected and interval CaP cases. RESULTS: Of the 728 men with a PSA level of 4.0 mg/mL or more who underwent biopsy at initial screening, 553 were eligible for a second screening visit after 4 years. Of these, 272 (49.2%) actually underwent screening. Eighteen CaP cases were detected with 217 biopsies, indicated by a PSA level of 3.0 ng/mL or more (PPV 8.3%). Eight interval cases were identified by linking to the Cancer Registry. These 26 cases would have increased the PPV and detection rate of the initial screening round from 36.1% to 39.7% and from 3.8% to 4.2%, respectively. Most of these cases (23 of 26 or 88.5%) were organ confined and amenable to potentially curative treatment. CONCLUSIONS: Although the results of this study may have been biased by the low rate of availability/eligibility of participants for rescreening (after 4 years), the proportion of cancers detected after a previous lateral sextant biopsy indicated by a PSA value of 4.0 mg/mL or more (PPV 8.3%) fell far short of the overall PPV at rescreening (PPV 20%). The features of most cancers that were possibly missed during the first round allowed a potentially curative approach. The ERSPC study group found no reason to change the ERSPC protocol. PMID- 15134976 TI - Pilot study to explore effects of low-fat, flaxseed-supplemented diet on proliferation of benign prostatic epithelium and prostate-specific antigen. AB - OBJECTIVES: Dietary factors may influence the prostate and have an impact on prostatic growth and disease. A small number of studies have suggested that flaxseed-supplemented, fat-restricted diets may thwart prostate cancer growth in both animals and humans. Unknown, however, is the potential effect of such a diet on benign prostatic epithelium. METHODS: We undertook a pilot study to explore whether a flaxseed-supplemented, fat-restricted diet affects the proliferation rates in benign epithelium. We also explored the effects on circulating levels of prostate-specific antigen (PSA), total testosterone, and cholesterol. Fifteen men who were scheduled to undergo repeat prostate biopsy were instructed to follow a low-fat (less than 20% kcal), flaxseed-supplemented (30 g/day) diet and were provided with a supply of flaxseed to last throughout the 6-month intervention period. The PSA, total testosterone, and cholesterol levels were determined at baseline and at 6 months of follow-up. Reports from the original and repeat biopsies were compared, and proliferation (MIB-1) rates were quantified in the benign prostatic epithelium. RESULTS: Statistically significant decreases in PSA (8.47 +/- 3.82 to 5.72 +/- 3.16 ng/mL; P = 0.0002) and cholesterol (241.1 +/- 30.8 to 213.3 +/- 51.2 mg/dL; P = 0.012) were observed. No statistically significant change was seen in total testosterone (434.5 +/- 143.6 to 428.3 +/- 92.5 ng/dL). Although 6-month repeat biopsies were not performed in 2 cases because of PSA normalization, of the 13 men who underwent repeat biopsy, the proliferation rates in the benign epithelium decreased significantly from 0.022 +/- 0.027 at baseline to 0.007 +/- 0.014 at 6 months of follow-up (P = 0.0168). CONCLUSIONS: These pilot data suggest that a flaxseed-supplemented, fat restricted diet may affect the biology of the prostate and associated biomarkers. A randomized controlled trial is needed to determine whether flaxseed supplementation, a low-fat diet, or a combination of the two regimens may be of use in controlling overall prostatic growth. PMID- 15134977 TI - Serum BPSA outperforms both total PSA and free PSA as a predictor of prostatic enlargement in men without prostate cancer. AB - OBJECTIVES: To determine whether the serum concentration of BPSA, a distinct form of free prostate-specific antigen (PSA) enriched in the nodular transition zone (TZ) tissue of benign prostatic hyperplasia (BPH), can predict TZ volume and diagnose BPH-associated prostatic enlargement in patients without prostate cancer. METHODS: We studied 91 consecutive patients without prostate cancer who underwent a 10-core or greater biopsy of the prostate. The associations between prostate volume, age, International Prostate Symptom Score, and serum concentrations of PSA, free PSA, and BPSA were evaluated by receiver operating characteristic curve and linear and binary logistic regression analyses. RESULTS: BPSA and free PSA showed stronger correlations with both age (BPSA = 0.38, free PSA = 0.40, PSA = 0.24) and TZ volume (BPSA = 0.67, free PSA = 0.64, PSA = 0.55) than did PSA. The percent free PSA had no statistically significant correlation with TZ volume (P = 0.08). Subtraction of BPSA from free PSA reduced its correlation with TZ volume to below that of PSA (from 0.64 to 0.48). Linear regression analyses showed that, unlike PSA, both BPSA and free PSA displayed an age-independent relationship to TZ volume. The receiver operating characteristic curve (for TZ greater than 30 cm3) and binary logistic regression analyses showed that BPSA (area under the curve = 0.844) outperformed both free PSA (area under the curve = 0.799) and PSA (area under the curve = 0.749) in its ability to predict clinically significant TZ enlargement. CONCLUSIONS: In patients without prostate cancer, the serum concentration of BPSA displayed an age-independent, log-linear relationship to TZ volume and was a better predictor of prostatic enlargement than either PSA or free PSA. BPSA may also predict clinical parameters of BPH and is under evaluation as a marker of BPH progression and response to therapy. PMID- 15134980 TI - Selenium levels of patients with newly diagnosed prostate cancer compared with control group. AB - OBJECTIVES: To compare the toenail selenium level of patients with prostate cancer with that of controls. A tumor-protective effect of selenium has been discussed for many years. Published data about the influence of the selenium level in the plasma and toenails are controversial. METHODS: Our prospective study included 150 patients. Of these, 70 patients had recently diagnosed prostate cancer (cancer group) and 80 patients had no evidence of prostate cancer (negative digital rectal examination and prostate-specific antigen level within age-specific range or negative biopsies) or any other malignant disease (control group). Toenail clippings exhibit selenium levels for up to 1 year. Both groups were compared by assessing their selenium levels using inductively coupled plasma dynamic reaction cell mass spectrometry after microwave-assisted closed vessel digestion. The results of the selenium levels in both groups were statistically compared using the Wilcoxon test. Possible correlations between the selenium level and age, body mass index, smoking habits, and prostate cancer were analyzed by multiple regression analysis. RESULTS: The median age of the cancer and control group was 65 and 69 years, respectively. The median toenail selenium level in the cancer and control group was 528 ng/g (range 393 to 4274) and 502 ng/g (range 201 to 831), respectively (P value not statistically significant). Furthermore, no correlation was found between selenium level and age, body mass index, or smoking status. CONCLUSIONS: The patients in our study with newly diagnosed and not yet treated prostate cancer did not have different toenail selenium levels than controls. Body selenium levels may not influence prostate cancer incidence. A protective effect of selenium for prostate cancer seems questionable. PMID- 15134981 TI - Influence of androgen deprivation therapy on volume of anatomic zones of prostate in patients with prostate cancer using magnetic resonance imaging. AB - OBJECTIVES: To clarify the influence of androgen deprivation therapy (ADT) on the volume of the anatomic zones of the prostate and the tissue component of the transition zone. METHODS: Thirty patients with prostate cancer arising and localizing in the peripheral zone were enrolled in this study. The volume of anatomic zones was measured using magnetic resonance imaging before and 6 months after ADT (castration and flutamide). Binary images were constructed to analyze the tissue components of the transition zone. RESULTS: The volumes after ADT in the whole prostate, transition zone, and peripheral zone were significantly smaller than the corresponding volumes before ADT (P <0.001). A small correlation was found between the reduction rates of the transition zone after therapy and the stromal rates of the transition zone (r(s) = 0.375, P = 0.041). CONCLUSIONS: The present results revealed that, not only the epithelial region in the transition zone, but also the stromal region, was sensitive to androgen and the volume of the transition zone was reduced during ADT regardless of its histologic components. PMID- 15134982 TI - Recurrent prostate cancer after external beam radiotherapy: value of contrast enhanced dynamic MRI in localizing intraprostatic tumor--correlation with biopsy findings. AB - OBJECTIVES: To assess the accuracy and interobserver variability of T2-weighted (T2W) and contrast-enhanced dynamic (CE-Dyn) magnetic resonance imaging (MRI) in predicting the results of transrectal biopsy in patients with suspected recurrent prostate cancer after external beam radiotherapy. METHODS: A total of 22 patients with increasing prostate-specific antigen levels after external beam radiotherapy for prostate cancer underwent T2W and CE-Dyn MRI of the prostate. The CE-Dyn sequence (acquisition time 30 seconds) was repeated three times after the injection of gadolinium. All patients underwent subsequent transrectal biopsy. Three independent readers interpreted the MRI scans. The MRI and biopsy results were correlated in 10 prostate sectors (the sextants of the peripheral zone, the two transitional zones, and the two seminal vesicles). RESULTS: Biopsy cores were obtained in 147 prostate sectors. Of these, 63 were positive for cancer in 19 patients. On the T2W images, the three readers interpreted as positive for cancer 15, 15, and 13 of the 19 patients showing cancer at biopsy. They interpreted as negative 3, 0, and 1 of the 3 patients showing no cancer at biopsy. On CE-Dyn images, the three readers correctly classified all the patients as positive or negative for cancer. The T2W and CE-Dyn MRI findings were concordant with biopsy results in, respectively, 81 to 95 and 107 to 117 prostate sectors (P <0.001 and P <0.01 for readers 1 and 2 and was nonsignificant for reader 3). The interobserver agreement was better for CE-Dyn images (kappa = 0.63 to 0.70) than for the T2W images (kappa = 0.18 to 0.39). The MRI-calculated tumor volumes and the mean biopsy core invasion rates were significantly correlated on the CE-Dyn images for all readers. They correlated significantly on T2W images only for one reader. CONCLUSIONS: CE-Dyn MRI depicts the intraprostatic distribution of recurrent cancer after external beam radiotherapy more accurately and with less interobserver variability than T2W MRI. PMID- 15134983 TI - Is the efficacy of hormonal therapy affected by lymph node status? data from the bicalutamide (Casodex) Early Prostate Cancer program. AB - OBJECTIVES: To report an exploratory subgroup analysis assessing the extent to which the overall benefit found in the Early Prostate Cancer program is dependent on lymph node status at randomization. The program is ongoing, and the overall survival data are immature. The first combined analysis of the bicalutamide (Casodex) Early Prostate Cancer program at 3 years' median follow-up showed that bicalutamide, 150 mg once daily, plus standard care (radical prostatectomy, radiotherapy, or watchful waiting), significantly reduced the risk of objective progression and prostate-specific antigen (PSA) doubling in patients with localized/locally advanced prostate cancer. METHODS: Men (n = 8113) with localized/locally advanced disease received bicalutamide 150 mg or placebo once daily, plus standard care. The time to event data (objective progression, PSA doubling) was analyzed by lymph node status at randomization. RESULTS: Compared with standard care alone, bicalutamide significantly reduced the risk of objective progression, irrespective of lymph node status, with the most pronounced reduction in patients with N+ (hazard ratio [HR] 0.29; 95% confidence interval [CI] 0.15 to 0.56) compared with those with N0 (HR 0.59; 95% CI 0.48 to 0.73) and Nx (HR 0.60; 95% CI 0.50 to 0.72) disease. The largest decrease in risk of PSA doubling with bicalutamide was observed in N+ disease (HR 0.16; 95% CI 0.09 to 0.29), with significantly reduced risks seen in N0 (HR 0.45; 95% CI 0.40 to 0.51) and Nx (HR 0.38; 95% CI 0.33 to 0.44) disease. CONCLUSIONS: The greatest reduction in the risk of objective progression and PSA doubling with bicalutamide was seen in patients with N+ disease. However, bicalutamide also provided a statistically significant benefit in those with N0 and Nx disease. PMID- 15134984 TI - Imatinib mesylate and zoledronic acid in androgen-independent prostate cancer. AB - OBJECTIVES: To determine the safety and efficacy of zoledronic acid (Zometa) combined with imatinib mesylate (Gleevec) in patients with bone pain due to androgen-independent prostate cancer. METHODS: Fifteen patients were treated with zoledronic acid 4 mg intravenously every 28 days and imatinib mesylate 400 mg/day. The pain response, defined as a 2-point reduction in the Present Pain Intensity Scale or normalization if the initial score was 1, was the primary endpoint. Secondary endpoints included palliative response, prostate-specific antigen response, measurable disease response, time to progression, impact on quality of life, decrease in markers of bone turnover, and tolerability of the drug combination. RESULTS: The study was stopped early because of a lack of activity. No palliative or clinical activity was detected for the combination, and no prostate-specific antigen responses were observed. The median time to progression was 4 weeks (95% confidence interval 3 to 5), and the median duration of treatment was 8 weeks (range 1.6 to 16.7). The median overall survival was 54 weeks (95% confidence interval 18 to 90). Therapy was associated with a reduction in urine N-telopeptides and a trend toward a reduction in serum osteocalcin, but no change occurred in bone-specific alkaline phosphatase. CONCLUSIONS: In this patient population, imatinib mesylate and zoledronic acid produced no prostate specific antigen responses and had no palliative or clinical activity. PMID- 15134985 TI - Samarium-153-Lexidronam complex for treatment of painful bone metastases in hormone-refractory prostate cancer. AB - OBJECTIVES: A Phase III randomized trial was designed to assess the effectiveness of samarium-153 (153Sm)-lexidronam for palliation of bone pain in patients with hormone-refractory prostate cancer. METHODS: A total of 152 men with hormone refractory prostate cancer and painful bone metastases were enrolled in a prospective, randomized, double-blind trial comparing radioactive (153Sm) versus nonradioactive (152Sm) lexidronam complexes. Patients were randomized (2:1) to the radioactive (153Sm) agent. Patient diaries recording daily pain and analgesic use were completed during a planned 16-week evaluation period. Nonresponders were informed of the treatment received after 4 weeks of treatment and, if initially treated with placebo, were allowed to receive 153Sm-lexidronam in an open-label fashion. Pain was measured using validated patient-derived visual analog scales and pain descriptor scales. RESULTS: 153Sm-lexidronam had positive effects on measures of pain relief compared with placebo within 1 to 2 weeks. Reductions in opioid use were recorded at weeks 3 and 4. Because nonresponders were unblinded at week 4, statistical comparisons between the arms beyond week 4 were not possible. Mild, transient bone marrow suppression was the only adverse event associated with 153Sm-lexidronam administration. The mean nadir white blood cell and platelet count (3 to 4 weeks after treatment) was 3800/microL and 127,000/microL, respectively. Counts recovered to baseline after approximately 8 weeks. No grade 4 decreases in either platelets or white bloods cells were documented. CONCLUSIONS: These findings demonstrate that 1 mCi/kg 153Sm lexidronam is both safe and effective for the palliation of painful bone metastases in patients with hormone-refractory prostate cancer. PMID- 15134986 TI - Neoadjuvant hormonal therapy impairs sexual outcome among younger men who undergo external beam radiotherapy for localized prostate cancer. AB - OBJECTIVES: To evaluate the effects of treatment factors and other covariates on sexual health-related quality of life (HRQOL) after radiotherapy (RT) for prostate cancer. The effects of clinical and treatment factors on sexual health after external beam RT have not been fully characterized by patient-reported, validated questionnaires. METHODS: A total of 259 subjects (147 patients who had undergone RT for prostate cancer and 112 age-matched controls) participated in a cross-sectional assessment of HRQOL using the validated expanded prostate cancer index to measure patient-reported sexual function and bother. Multivariable models were used to determine the association of 13 prospectively measured clinical and treatment factors with post-RT sexual HRQOL. RESULTS: Increasing age, time since RT, and use of neoadjuvant hormonal therapy (NHT) were independently associated with sexual function (P <0.01) after RT. The effects of NHT on sexual HRQOL were most notable among the youngest (younger than 67 years) patients and among those closest to treatment. These effects may be conceptualized by examining the distribution of responses to individual Expanded Prostate Cancer Index Composite questions. For example, among the youngest patients, those treated with NHT were less than half as likely to report a fair ability to have erections compared with hormone-naive men. CONCLUSIONS: These patient-reported data, elicited using a validated HRQOL instrument, indicate that older age and NHT significantly and independently hinder post-RT sexual HRQOL outcome among men with localized prostate cancer and provide a framework for counseling patients regarding long-term sexual outcome on the basis of treatment plan and age. PMID- 15134987 TI - Use of intraurethral alprostadil in patients not responding to sildenafil citrate. AB - OBJECTIVES: To determine whether intraurethral alprostadil would be an effective alternative for men with erectile dysfunction who did not respond adequately to sildenafil citrate but desired minimally invasive treatment. METHODS: A total of 44 male patients aged 41 to 74 years with erectile dysfunction refractory to treatment with sildenafil citrate were enrolled in this study. Of the 44 patients, 10 had undergone prior radical retropubic prostatectomy. The patients were evaluated for subjective improvement in an office setting and completed the Sexual Health Inventory for Men questionnaire as an objective assessment of improved erectile ability. Success was defined as subjective improvement in erectile function, as well as an improved Sexual Health Inventory for Men score. RESULTS: Of the 44 men, 13 (29.5%) responded successfully to intraurethral alprostadil, with a follow-up ranging from 2 to 15 months. The remaining 31 men had no response (n = 28, 90%), refused escalating doses (n = 2, 7%), or were lost to follow-up (n = 1, 3%). In the subgroup of 10 men with prior radical retropubic prostatectomy, 5 (50%) reported success with intraurethral alprostadil (500 microg in 2 patients and 1000 microg in 3 patients). CONCLUSIONS: Although sildenafil citrate remains the most common initial therapy in men with erectile dysfunction, intraurethral alprostadil may be a reasonable treatment option for sildenafil nonresponders. This may be especially true in men having undergone prior radical retropubic prostatectomy. PMID- 15134988 TI - Placebo-controlled study on efficacy and safety of daily apomorphine SL intake in premenopausal women affected by hypoactive sexual desire disorder and sexual arousal disorder. AB - OBJECTIVES: To verify whether apomorphine SL is effective in premenopausal women affected by arousal disorder with hypoactive sexual desire disorder. METHODS: Sixty-two volunteer women, aged 26 to 45 years, affected by arousal disorders and hypoactive sexual desire disorder participated in the study, which consisted of two parts. The first was 4 weeks of a taken-as-needed, open-label, dose escalation regimen starting at 2 or 3 mg of apomorphine SL. The second part of the study was for the nonresponders, who were randomly allocated to treatment in one of six possible sequences of three 2-week double-blind, crossover study periods with apomorphine 2 mg or 3 mg, washout, and placebo. Efficacy was assessed with the Personal Experiences Questionnaire using the 5-point Likert scale. The Personal Experiences Questionnaire quantified subjective arousal, desire, orgasm, enjoyment, and frequency of sexual relationships. RESULTS: Fifty women completed the 4-week "as required" drug regimen, and 6 of them benefited from this treatment regimen (P <0.05). The 44 women who reported no change with respect to baseline participated in the double-blind crossover part of the study. Six women stopped taking the drug because of a fear of problems (two each during the 2-mg apomorphine, 3-mg apomorphine, and placebo periods). Adverse events were mild or moderate, both occurred during the "as required" part (10 patients) and during daily usage (6 patients) and were mainly nausea, vomiting, dizziness, or headache. However, during the placebo period, 2 women had adverse events, mainly headache. The daily intake of the drug was effective with both the 2-mg and 3-mg dosages compared with placebo for arousal and desire (P <0.05). The effects of 3 mg of apomorphine were better than those obtained with 2 mg (P <0.05). The orgasm, enjoyment, and satisfied by frequency scores improved during treatment with daily apomorphine compared with baseline and placebo. CONCLUSIONS: The results of our study suggest that daily apomorphine SL may improve the sexual life of women affected by sexual difficulties. Additional studies are needed to define the daily use of apomorphine SL in large subgroups of women on the basis of etiology and the severity of sexual dysfunction. PMID- 15134989 TI - Efficacy and factors associated with successful outcome of sildenafil citrate use for erectile dysfunction after radical prostatectomy. AB - OBJECTIVES: To assess the efficacy and factors associated with successful treatment of sildenafil citrate for erectile dysfunction after radical prostatectomy (RP). METHODS: Of the 470 patients who underwent RP at our institution between July 1998 and January 2000, 227 (48%) sought treatment for erectile dysfunction, and 174 (37%) were prescribed sildenafil citrate. The starting dose was 50 mg, which was increased to 100 mg if the patient did not have a positive response. Of the 174 patients, 104 (59.8%) had undergone a bilateral nerve-sparing (NS) procedure, 28 (16.1%) had undergone a unilateral NS procedure, and 42 (24.1%) had undergone a non-NS procedure. Erectile function was assessed by the abridged five-item version of the International Index of Erectile Function questionnaire, referred to as the Sexual Health Inventory for Men (SHIM), at baseline and 1 year after sildenafil use. The patients' charts were retrospectively reviewed to find factors associated with a successful outcome, which was defined as successful vaginal intercourse. Association with success was assessed by chi-square analysis and the Cochran Armitage test for trend. Bonferroni correction for multiple comparisons was used, with an overall significance level of 0.05 for each factor assessed. RESULTS: The mean age was 60.1 +/- 6.25 years, and the mean interval from RP to drug use was 3 months. After treatment with sildenafil, 100 (57%) of 174 patients responded to the drug: 79 (76%) of 104 in the bilateral NS group, 15 (53.5%) of 28 in the unilateral NS group, and 6 (14.2%) of 42 in the non-NS group. SHIM analysis showed that the magnitude of the improvement was greater in the bilateral NS group (19.97 +/- 1.12) than in the unilateral NS (15.89 +/- 3.38) or non-NS (10.06 +/- 2.0) groups (P <0.020). Four factors were significantly associated statistically with a successful outcome: the presence of at least one neurovascular bundle, a preoperative SHIM score of 15 or greater, age 65 years old or younger, and interval from RP to drug use of more than 6 months (P <0.001). CONCLUSIONS: The efficacy of sildenafil citrate after RP correlated with the degree of neurovascular bundle preservation, preoperative erectile function status, age, and interval before starting treatment. PMID- 15134990 TI - Reliability of voiding cystourethrography to detect urethral obstruction in boys. AB - OBJECTIVES: To evaluate the reliability of voiding cystourethrography (VCUG) to diagnose infravesical obstruction in boys. METHODS: Hard copies of the VCUG findings of 72 boys were assessed by two pediatric radiologists and two pediatric urologists. The investigators were instructed to consider six items related to infravesical obstruction: vesicoureteral reflux, bladder wall thickness, bladder diverticulum, abnormal prostatic urethra, visible urethral obstruction, and obstruction in the sphincter area. Also, a scoring system was developed using these six items. Agreement among the four investigators for evaluation of the items on VCUG and for the scoring system was assessed using the kappa statistic. All boys underwent urethrocystoscopy, and the endoscopic findings were compared with VCUG results. Odds ratios were calculated for the results of VCUG for each investigator to predict the chance of cystoscopic infravesical obstruction. RESULTS: Agreement among observers for vesicoureteral reflux and bladder diverticulum was good (kappa values for paired observers of 0.82 and 0.79). Agreement for bladder wall thickness, abnormal prostatic urethra, visible urethral obstruction, obstruction in the sphincter area, and the scoring system was poor (kappa values of 0.08, 0.35, 0.33, 0.26, and 0.33, respectively). Consequently, the results of VCUG could not predict for endoscopic infravesical obstruction accurately, although substantial differences occurred among investigators. Items on which investigators reached good agreement were negatively related to the risk of having infravesical obstruction. CONCLUSIONS: Because agreement among investigators in the assessment of most items on VCUG was poor and because for the items with good agreement, the predictive power was poor, the current clinical use of VCUG for diagnosing infravesical obstruction needs reevaluation. PMID- 15134993 TI - Extraperitoneal bilateral cutaneous ureterostomy with midline stoma for palliation of pelvic cancer. AB - In a subset of patients with intractable symptoms from incurable pelvic cancer, palliative cystectomy and urinary diversion with intestinal segments may not be feasible. The described technique, in which a midline stoma was constructed and dissection was entirely extraperitoneal, resulted in symptom relief in 8 patients. PMID- 15134994 TI - A simple and reliable hemostatic technique during partial nephrectomy. AB - INTRODUCTION: To report our experience with a refined technique for hemostasis that obviates the need for vascular control and closure of the collecting system in partial nephrectomy. TECHNICAL CONSIDERATIONS: Four to five sutures, 2 cm apart, are placed 0.5 cm from the anticipated parenchymal incision border, using a specially designed, blunt-tip, straight needle with folded 2-0 Vicryl thread. The needle is removed, leaving the Vicryl thread with the loop on one side of the kidney and two free ends on the other side. A 1.0-cm-wide Vicryl mesh strip is passed circumferentially through the loops and between the free ends on each side, tension is applied on the strip during knotting of the free ends of the thread, and the tissue is incised. No additional hemostatic sutures are necessary. No attempt is made to identify and close the open collecting system. Vascular clamping and surface cooling are avoided. Sixty-one patients have undergone this technique since 1987: initially, for complicated nephrolithiasis (n = 15), localized purulent kidney disease (n = 4), trauma (n = 3), congenital anomalies (n = 2), and resection of horseshoe kidney (n = 6) and, recently, for peripherally located renal tumor (n = 31). Upper pole resection was performed in 11 patients, lower pole resection in 45, and middle segment resection in 5. The blood loss was minimal, with only 1 patient developing gross hematuria that resolved after conservative treatment. No other complications occurred. CONCLUSIONS: A simple and easily performed hemostatic method suitable for peripherally located and, particularly, polar renal tumors is described. The Vicryl mesh strip prevents tears of the parenchymal sutures and ensures good hemostasis without closing the collecting system separately. PMID- 15134995 TI - Isolated bilateral renal mucormycosis. PMID- 15134996 TI - Transitional cell carcinoma of the bladder metastatic to the penis. AB - Transitional cell carcinoma of the bladder has the potential to metastasize to multiple organs, and the vascularity of the penis makes it a potential site for hematogenous metastases. We present 2 cases of transitional cell carcinoma with metastases to the penis, with a review of the published reports involving penile metastases. We also discuss the presentation of penile metastases and relevant management issues in these patients who typically have quite advanced disease at presentation. PMID- 15134997 TI - Benign renal mesenchymoma in the pediatric age group: a novel pathologic and karyotype entity. AB - Benign mesenchymal renal tumors are extremely rare in the pediatric age group. We report a case of a benign renal tumor composed of smooth muscle cells, adipose tissue, and areas of cartilaginous differentiation and expressing a 46,XX, t(8;10)(q21;q24) karyotype in a 13-year-old girl. Although some pediatric renal tumors show a degree of heterologous differentiation, none of them exclusively consist of these three well-differentiated mesenchymal components. The unique features of the present case were further confirmed by the karyotype changes, which to our knowledge, have never been described before in a pediatric renal tumor. PMID- 15134998 TI - Testicular involvement in disseminated fungal infection by Pseudallescheria boydii. AB - We present an immunocompromised patient with a painless testicular mass. Usually, this mass is suspicious for testicular cancer; however, a fungal abscess from Pseudallescheria boydii was found. The patient eventually died from disseminating infection. This is the first such report of a testicular infection involving P. boydii. Early diagnosis and treatment are paramount because of the poor outcome with disseminated disease. PMID- 15134999 TI - Leech therapy in penile replantation: a case of recurrent penile self-amputation. AB - Penile amputation is a rare urologic trauma for which immediate surgical replantation is indicated. Microsurgical techniques can reduce skin and graft loss complications; nonetheless, such complications are still highly prevalent. We report a case of self-inflicted penile amputation and describe a nonmicrosurgical technique for replantation. To improve postoperative edema due to venous congestion, we applied medicinal leeches to the penis. The edema quickly resolved, but overlying skin loss occurred, which required superficial debridement. At follow-up the patient had glans re-epithelialization with normal voiding, sensation, and erections. To our knowledge, this is the first reported case of nonmicrosurgical penile replantation with leech therapy. PMID- 15135000 TI - Bilateral testicular adrenal rests after bilateral adrenalectomies in a cushingoid patient with von Hippel-Lindau disease. AB - We report a case of bilateral testicular masses in a 25-year-old man with von Hippel-Lindau disease presenting with cushingoid symptoms. His medical history was significant for bilateral adrenalectomies secondary to pheochromocytomas, and he began steroid therapy at that time. After exhaustive endocrinologic, radiographic, and physical examinations, the testicular masses were postulated to be active adrenal rest tissue. Bilateral testicular venous sampling found elevated glucocorticoids that were responsive to dexamethasone suppression, which confirmed the testicular masses as testicular adrenal rests without the need for surgical intervention. Successful conservative management consisted of appropriate steroid manipulation and radiographic evaluation and resulted in the resolution of presenting symptoms, a decrease in size of the bilateral testicular masses, and testicular conservation in this young man. PMID- 15135001 TI - Appendiceal cystadenoma mimicking a cystic renal mass. AB - We report the first case of appendiceal mucinous cystadenoma associated with the kidney. A 57-year-old man with chronic renal failure presented with a right renal mass. During laparoscopic nephrectomy, frozen analysis of a tubular structure extending from the mass showed appendiceal tissue. The mass, appendix, and what was believed to be the right kidney were removed. Pathologic examination demonstrated mucinous cystadenoma of the appendix. No renal tissue was identified, but the patient refused further treatment. This case demonstrates that mucinous cystadenomas arising from retroperitoneal structures might mimic renal neoplasms and should be considered in the differential diagnosis of cystic renal masses. PMID- 15135002 TI - Prenatal puncture of a unilateral hydronephrosis leading to fetal urinoma and postnatal nephrectomy. AB - Fetal pelvicaliceal dilatation due to ureteropelvic junction obstruction is the most common cause of antenatal hydronephrosis; it rarely leads to a spontaneous rupture resulting in urinoma formation. Antenatal intervention has been recommended only in those cases of large urinomas that seem to interfere with the function of other organ systems (eg, pulmonary hypoplasia secondary to diaphragmatic elevation). We report the case of a fetal intervention (transuterine puncture) in a unilateral massive hydronephrosis leading to a perirenal urinoma and the preterm birth of a female infant. Postnatally, mechanical ventilation and oxygen were required, as was forced percutaneous urinoma drainage. Evaluation revealed a fistula formation between the perirenal space and the kidney's collecting system, possibly due to the fetal intervention. Unfortunately the kidney function was very poor, and surgery to remove the impaired kidney and the urinoma was performed. We discuss the possible effects of fetal intervention in cases of obstructive uropathy and the postnatal risks associated with it. PMID- 15135003 TI - Retroperitoneoscopic ureterocalicostomy for congenital proximal ureteral stenosis. AB - We report the first case of retroperitoneoscopic ureterocalicostomy in a 17-year old male patient with severe left hydronephrosis caused by a long congenital upper ureteral stenosis. With a retroperitoneoscopic approach, the stenotic segment was resected, the thin renal parenchyma overlying the lower calyx was fenestrated by a modest excision, and the proximal ureter was anastomosed to the lower pole in an end-to-end manner. At 2 years postoperatively, the patient was asymptomatic, with a significant reduction in hydronephrosis and a patent upper ureter. Retroperitoneoscopic ureterocalicostomy is technically feasible and can provide long-term successful reconstruction of a complicated ureteropelvic junction obstruction. PMID- 15135004 TI - Biologic feature of prostatic hyperplasia developed in spontaneously hypertensive rats. AB - OBJECTIVES: To investigate alterations in cell kinetic and cellular composition in prostatic hyperplasia developed in spontaneously hypertensive (SH) rats from 15 to 54 weeks of age. METHODS: Male SH rats (n = 20) and their normotensive counterparts, Wistar-Kyoto rats (n = 20), were studied. At 15, 29, 40, and 54 weeks of age, the ventral prostate was obtained to evaluate (a) cell proliferation by the proliferation index using proliferating cell nuclear antigen immunostaining and (b) apoptosis by the apoptotic index using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling assay. The percentage of area density of smooth muscle and fibroblastic tissues was determined with a computerized image analysis system after alpha-actin and vimentin immunostaining, respectively. RESULTS: Compared with the data in the Wistar-Kyoto rats during the observation period, a significantly increased proliferation index in both epithelium and stroma (P <0.01) and a slightly increased apoptotic index in the epithelium without apoptotic cells in the stroma, with an imbalance in favor of cell proliferation, were noted in the prostatic hyperplasia of the SH rats. The value of the proliferation index was greatly increased in both the epithelium and the stroma of the SH rats between 15 and 29 weeks (P <0.05) but continued relatively steady from 29 to 54 weeks. A significantly increased percentage of smooth muscle area (P <0.01) and a much greater percentage of area density of smooth muscle than that of fibroblastic tissues (P <0.01) were observed in the prostatic hyperplasia of the SH rats. CONCLUSIONS: The prostatic hyperplasia of SH rats may develop from both epithelial and stromal growth and could ultimately be promoted by an imbalance in favor of proliferative activity. PMID- 15135005 TI - Clinical evaluation of p53 mutations in urothelial carcinoma by IHC and FASAY. AB - OBJECTIVES: To assess the clinical benefit of the methods of detection of p53 mutations in human urothelial carcinoma. METHODS: A total of 75 surgical specimens of urothelial carcinoma were analyzed using a yeast functional assay (FASAY) and immunohistochemistry (IHC), in combination with sequencing analysis. RESULTS: Of the 75 specimens, 24 (32.0%) were positive (mutant) by FASAY and 23 (30.7%) were positive by IHC. The sequencing analysis confirmed that all 24 FASAY positive tumors harbored mutations, and no mutations were detected in any FASAY negative tumors. In contrast, nuclear accumulation of p53 protein was detected in 9 (17.6%) of 51 tumors with no mutation, and 10 (41.7%) of 24 tumors with mutation showed no positive staining on IHC. The mutations detected by FASAY and IHC were both associated with stage and grade, but null mutations of p53 were not associated with stage. Concerning chemosensitivity, 6 (85.7%) of 7 responders harbored p53 missense mutations in at least one allele (P = 0.01), and only 4 (57.1%) were judged positive by IHC (P = 0.13). CONCLUSIONS: FASAY is more accurate than IHC in detecting the various types of p53 mutations, suggesting that a comprehensive approach for the detection of p53 mutations may be essential to elucidate their clinical significance. PMID- 15135006 TI - Modulation by desmopressin of neuronal activity in brainstem micturition center. AB - OBJECTIVES: To examine the effect of desmopressin (DDAVP) on bladder contraction and on the neurons that fire in relation to spontaneous bladder contraction (bladder-related neurons) in and around Barrington's nucleus, the micturition center. DDAVP is used for the treatment of nocturnal enuresis because of its antidiuretic action, but the mechanism of this action has not been proved. METHODS: Urethane-anesthetized Sprague-Dawley male rats (n = 20) were used. DDAVP was infused intravenously or as an intracerebroventricular infusion into the lateral ventricle. RESULTS: We encountered three types of bladder-related neurons: those that fired before the start of the contraction (type E1), those that fired synchronous with the bladder contraction (type E2), and those that fired during bladder relaxation (type I). Intravenous infusion caused inhibition in three of five type E1 neurons, excitation in two of five type E2 neurons, and excitation (one neuron) and inhibition (one neuron) of four type I neurons. With intracerebroventricular infusion into the lateral ventricle, two of four type E1 neurons were inhibited, and one of seven type E2 neurons and three of four type I neurons were excited. Bladder contraction was suppressed in 4 of 12 rats by intravenous infusion and in 2 of 8 rats by intracerebroventricular infusion into the lateral ventricle. In all cases, when the bladder contraction was suppressed, an electroencephalogram of larger amplitude and slower frequency appeared. CONCLUSIONS: DDAVP seems to regulate bladder activity by affecting bladder related neurons in the micturition center. PMID- 15135007 TI - Testosterone treatment of human foreskin in a novel transplant model. AB - OBJECTIVES: To determine whether testosterone would increase angiogenesis in human foreskin, because of the known effects of testosterone on endothelial cells and vascular endothelial growth factor expression. Surgical management of complex hypospadias in a patient who has undergone multiple procedures is technically very challenging. It is possible that therapy to increase the blood supply to this tissue could be beneficial. METHODS: We used a newly developed model of human skin transplantation, in which full-thickness human foreskin is transplanted subcutaneously onto the dorsum of a nude rat. At 10 days after transplantation, tissue was treated with either testosterone gel or vehicle control. After an additional 7 days, the tissue was harvested, embedded in paraffin, stained for factor VIII to assess vascularity, and examined histologically. RESULTS: The testosterone-treated tissue demonstrated increased factor VIII staining (31.14 +/- 1.53 vessels per high-power field) compared with the control group (18.25 +/- 2.3 vessels per high-power field; P <0.0005). Histologic analysis revealed less collagen in the testosterone-treated group compared with the control group. CONCLUSIONS: Treatment of human foreskin with testosterone, in a transplant model, increased vascularity and decreased early fibrosis. Testosterone treatment may improve the surgical management of complex hypospadias repair. PMID- 15135008 TI - Improvement in relaxation response in corpus cavernosum from trained rats. AB - OBJECTIVES: To evaluate the contractile and relaxing responses in rat corpus cavernosum (RCC) from rats after 8 weeks of run training, because erectile function is highly dependent on nitric oxide (NO) from nitrergic fibers or endothelium. Physical activity enhances NO production and improves endothelial function, with beneficial effects on cardiovascular disease. METHODS: The training program consisted of 8 weeks of run training, 5 days/wk, and each session lasted 60 minutes. The RCC was isolated, and concentration-response curves to NO, acetylcholine, sodium nitroprusside, phenylephrine, and endothelin were obtained. The excitatory and inhibitory effects of electrical field stimulation (2 to 32 Hz) were also evaluated. RESULTS: NO (0.1 to 100 microM) and sodium nitroprusside (0.01 to 1000 microM) produced a relaxing effect in RCC in a dose-dependent manner, with the maximal responses to NO (control 62% +/- 4%, trained 88% +/- 3%) and sodium nitroprusside (control 83% +/- 3%, trained 95% +/- 2%) significantly enhanced after 8 weeks of run training. However, acetylcholine induced relaxations were not affected by exercise. Similarly, electrical field stimulation-induced relaxations were significantly increased in RCC from trained rats at 2 Hz (control 2.4% +/- 0.3%, trained 4.2% +/- 0.5%) and 4 Hz (control 5.3% +/- 1.2%, trained 12.5% +/- 1.7%). The contractile sensitivity of RCC to phenylephrine (0.01 to 100 microM) and endothelin (0.01 to 100 nM) was not modified by training exercise. CONCLUSIONS: Our findings suggest that run training enhances functional responses in rat RCC that involves increases in the NO-cyclic guanosine monophosphate signaling pathway by endothelium-independent mechanisms that is not accompanied by changes in contractile sensitivity. PMID- 15135009 TI - Classical conditioning and conditionability of insulin and glucose effects in healthy humans. AB - We examined whether the effects of intravenously injected insulin and glucose (the physiological endogenous insulin production stimulus) could be classically conditioned in healthy humans. We expected a conditioned blood glucose decrease to a conditioned stimulus (CS) previously paired with insulin and an, albeit lower, blood glucose decrease to a CS paired with glucose injection. In addition, we analyzed glucoregulatory hormone and symptom conditionability. Thirty healthy males were divided into three groups and were given the CS and an intravenous injection of either insulin (0.05 IU/kg) in Group 1, glucose (15%, 0.5 g/kg) in Group 2, or placebo [physiological saline (0.9%)] in Group 3 during the acquisition phase on 4 days. All participants were given the olfactory CS (rosewood-peppermint smell) and placebo injection on Day 5 (test). On Day 5, the total blood glucose decrease tended to be higher in Group 1 than in Group 3 (P<.10), especially at CS presentation (P<.10) and previous unconditioned hypoglycemia time-point (P<.05). The conditioned blood glucose decrease was statistically nonsignificant in Group 2, but shortly after CS presentation, insulin level and blood glucose changes were negatively correlated in Groups 1 and 2 in contrast to positive correlation in Group 3. Furthermore, Group 1 showed an increase in noradrenaline (P<.05), a temporarily delayed increase in growth hormone (GH; P<.05), and an increase of autonomic and neuroglycopenic symptoms, reaching a medium and small effect size, respectively. Group 2 responded with an increase in cortisol (P<.01) and neuroglycopenic symptoms (P<.05) at the time point of the previous unconditioned blood glucose minimum. To conclude, the effects of exogenously applied insulin can be conditioned in a reliable way. In correspondence with the lower intensity of the unconditioned stimulus (US), conditioning effects with glucose-and, thus, endogenously produced insulin-are weaker but also reflect the actions of central insulin. Future studies will examine the diverse actions of insulin within the brain further. PMID- 15135010 TI - Validation of a stimulation protocol suited to the investigation of odor-taste interactions with fMRI. AB - A protocol suited to the investigations of odor-taste interactions in a functional magnetic resonance imaging (fMRI) scanner is described and results that confirm the validity of that protocol are presented. Stimuli were dissolved in water and presented to the mouth of the participant through plastic syringes and plastic tubes as 50 microl every 3 s for 18 s. Participants were lying on the back to simulate the conditions in a fMRI scanner and gave pleasantness and intensity ratings in response to each stimulus using a Labeled Magnitude Scale (LMS). Stimuli were three concentrations of two tastants (i.e., sucrose and NaCl) and two odorants [i.e., ethyl butyrate (fruity) and citral (citrus)] and all possible odor-taste mixtures. Results showed that additivity of intensity in odor taste mixtures was linear and did not depend on the pleasantness of the mixture. This study showed the feasibility of acquiring reliable psychophysical data with a stimulation device compatible with the fMRI environment and provided the foundation for the interpretation of future fMRI investigations of odor-taste interactions in flavor. PMID- 15135012 TI - Immobilization and cold stress affect sympatho-adrenomedullary system and pituitary-adrenocortical axis of rats exposed to long-term isolation and crowding. AB - Changes in plasma levels of noradrenaline (NA), adrenaline (A), adrenocorticotropic hormone (ACTH) and corticosterone (CORT), as well as in cytosol glucocorticoid receptor (GR) and heat shock protein 70 (Hsp 70) in hippocampus of adult rat males exposed to two long-term types of psychosocial stress, both under basal conditions and in response to immobilization and cold as heterotypic additional stressor were studied. Long-term isolation produced a significant elevation of basal plasma ACTH and CORT levels, but did not affect that of NA and A, while long-term crowding conditions did not elevate the basal plasma levels of these hormones. Long-term isolation of rats exposed to 2 h of immobilization or cold led to a significant elevation of plasma NA, A and CORT in comparison with the controls. Long-term crowding conditions and exposure of animals to immobilization or cold also resulted in an increased plasma NA, A and CORT levels, but to a lesser extent in comparison with the long-term isolation. At the same time, plasma ACTH was significantly more elevated in long-term crowded than in long-term isolated rats. Both kinds of long-term psychosocial stresses (isolation and crowding) had similar but less pronounced effects on cytosol GR and Hsp 70 concentrations in hippocampus comparing to acute immobilization and cold stress. It seems that long-term psychosocial stresses attenuate the effects of an additional stress on hippocampal GR and Hsp 70 concentrations. These data suggest that individual housing of rats appear to act as a stronger stressor than crowding conditions. When the animals suffering a long-term isolation were exposed to either acute immobilization or cold, a stronger activation of the sympatho-adrenomedullary system (SAS) was recorded in comparison with that found in the long-term crowded group subjected to short-term immobilization or cold. No significant differences in the activity of hypotalamo pituitary-adrenal (HPA) axis were observed between long-term isolated and long term crowded rats. PMID- 15135011 TI - Furosemide-induced food avoidance: evidence for a conditioned response. AB - Furosemide (Furo) is a potent natriuretic drug that is often used experimentally to investigate the brain mechanisms underlying salt appetite. Within this experimental paradigm, however, Furo also has anorectic activity that has received only modest attention. In Experiment 1 we varied two things administering a 10-mg dose of Furo in a single or a divided dose and preinjection exposure to a Na-free diet. In the 24 h after Furo, all four groups of rats reduced ingestion of Na-free diet. Both the division of the Furo dose and the preexposure to Na-free diet reduced the amount of food consumed even more than a single dose or continuous access to normal chow did. The fact that preexposure to Na-free diet increased the post-Furo anorexia implied an associative component to the phenomenon. Experiments 2 and 3 investigated the ability of Furo (2 and 10 mg) to serve as an unconditioned stimulus in taste aversion learning using 0.2 M sucrose as the conditioned stimulus. A saline (Sal) injection group served as control in both experiments. The results show that animals avoided sucrose when its ingestion was immediately followed by 10 mg Furo but not with 2 mg Furo or Sal. An aversion to sucrose did not develop when 10 mg Furo was administered the day prior to sucrose access. Thus, the suppressive effects of high-dose Furo on food intake might be due to a conditioned response. PMID- 15135013 TI - Behaviour in the elevated plus-maze predicts coping after subchronic mild stress in mice. AB - This study was aimed at investigating the coping style of mice subjected to a subchronic unpredictable mild stress procedure and its relationship to initial emotional reactivity. Two inbred strains of mice, the BALB/c ByJ and the C57BL/6 J, known to exhibit distinct emotionality, have been used. They were first observed in the elevated plus-maze and the free exploratory paradigm, each provides a separation of the population in high and low emotional mice. Half of the mice of each strain were then confronted to a 2-week subchronic unpredictable mild stress and tested for their responses in different behavioural situations (consumption of a palatable food, physical state, grooming behaviours and reactivity to a conflict situation). Mice were also tested in the light/dark procedure to assess the effect of the subchronic stress on emotional reactivity. First, a relationship between initial emotional reactivity in the elevated plus maze and behavioural coping style in response to stress was found, high emotional mice (i.e., BALB mice) displaying inhibited behaviours and less emotional mice (i.e., BL/6 mice) exhibiting few behavioural changes. Furthermore, emotional reactivity was increased in stressed mice compared with nonstressed ones. PMID- 15135014 TI - Attraction to male pheromones and sexual behaviour show different regulatory mechanisms in female mice. AB - In rodents, female sexual behaviour is under hormonal control. The attraction females show for male-derived nonvolatile chemicals (pheromones) can be regarded as the first step of this behaviour, but it is unknown whether this attraction is also modulated by sexual steroids. To test this possibility, ovariectomized adult female mice with no experience of chemical signals from adult males were randomly assigned to four groups that received oil (control), progesterone, estradiol (E) or estradiol+progesterone (E+P) injections, respectively. Females were then tested for their attraction to male-soiled bedding and, subsequently, for their proceptive behaviour when confronted to adult males. Females showed attraction to male-soiled bedding irrespective of the hormonal treatment, whereas only those females treated with E or E+P showed proceptive behaviour. Therefore, in contrast to proceptive and copulatory behaviour, the female attraction to male pheromones is independent of sexual steroids, thus indicating that those parts of the vomeronasal system involved in this attraction do not respond to steroids. In summary, sexual behaviour in female mice can be seen as a two-step process. First, females are attracted by male pheromones, a process which is independent of their hormonal status. After encountering the males, females show proceptive behaviour only in estrous, when fertilization is more likely. The attraction exerted by male sexual pheromones promotes female autostimulation that might ensure anticipatory endocrine changes leading to ovulation by the time of sexual intercourse. PMID- 15135015 TI - Olfactory sensitivity, learning and cognition in young adult and aged male Wistar rats. AB - Psychophysical experiments with male Wistar rats, ranging from 2 to more than 25 months old, revealed age-related differences in olfactory sensitivity. The highest sensitivities were found in rats 13 months old, and the lowest sensitivity was found in the group aged 25 months and older. Consequently, we considered the hypotheses that young rats will require less time and less trials than aged conspecifics to learn an olfactory discrimination task and that olfactory cognitive abilities will be reduced in older individuals. Rats were initially trained in an olfactometer using operant techniques to discriminate between the odor ethyl acetate (EA) and clean air. Next, young adult and 28-month old rats were tested on seven different go/no-go odor discrimination tasks. Aged rats performed as well as young adults did on all tasks and we conclude that, for a variety of odor discrimination problems, aged rats show no deterioration in learning ability. This is the first report on olfactory sensitivity, learning ability and cognition in Wistar rats that have passed the normal life span for this strain. Data show that the inability to learn and cognitive deficits do not necessarily develop with age. PMID- 15135016 TI - Long-term effects of the periadolescent environment on exploratory activity and aggressive behaviour in mice: social versus physical enrichment. AB - The aims of the present study were (i) to investigate the effects of environmental enrichment during periadolescence on different behavioural and neurochemical responses in male CD-1 mice at adulthood and (ii) to describe the relative role of the physical and social components of the enrichment in producing these effects. Thirty 5-day-old mice were randomly assigned to one of the following housing conditions lasting five consecutive days: (i) individually housed in a standard cage, (ii) housed in pairs in a standard cage, (iii) individually housed in a physically enriched cage, and (iv) housed in pairs in a physically enriched cage. At adulthood, 80 days after the enrichment exposure, the explorative behaviour in an open field, as well as the behaviour in agonistic encounters, was evaluated in association with the analysis of selected central (hypothalamic levels of nerve growth factor (NGF) and brain-derived growth factor(BDNF)) and peripheral (plasma corticosterone levels) biochemical parameters. The results show that the long-term effects of the physical and the social enrichment are different and not additive. In particular, while social enrichment by itself exerted very limited effects, physical enrichment decreased the exploratory activity and altered social behaviour. Mice housed in pairs in an enriched cage showed low activity levels in the open field, and they tended to become more frequently dominant, although showing a more affiliative and less aggressive social interaction strategy. Furthermore, they presented low levels of hypothalamic NGF and high levels of brain-derived growth factor, suggesting an important effect of the combination of social and physical enrichment on neurobehavioral markers of brain plasticity and on animal ability to cope with social challenges. PMID- 15135017 TI - Social categories in families of Mongolian gerbils. AB - Mongolian gerbils are a species of rodent in Asia living in families, although aggression occurs within these social units. The aim of this study was to determine the causes for the surge of intrafamily aggression and to characterize different types of animals. Due to behavioral measures, animals were assigned to distinct social categories and the consequences of social stress were analyzed on an individual level. Four families established from founder pairs were kept for up to 2.5 years in large enclosures. Social interactions within each family were recorded for at least 5 days per week throughout the long-term experiment and fecal samples were collected weekly for the determination of corticosterone concentration. Moreover, adrenals were weighed and histologically analyzed. Two main causes for the outbreak of aggression were distinguished: changes in family structure and female competition for reproduction. As a result of these aggression periods, it was possible to divide animals into social categories: (1) integrated family members (IFM), (2) founder pair animals (FPA) and (3) expelled family members (EFM). Integrated animals had the lightest adrenal glands [related to the fat-free mass (FFM)] and lowest corticosterone level in aggression periods. Founder animals were never attacked by other members, reproduced successfully and displayed the highest stress level in most of the measured parameters. EFM were attacked and excluded mainly by the founder females. Their stress level was intermediate. PMID- 15135018 TI - The rat exposure test: a model of mouse defensive behaviors. AB - In order to facilitate behavioral, and potentially pharmacological, analyses of risk assessment behaviors in mice, a rat exposure test (RET) was devised and evaluated. This test provides a home chamber connected via a tunnel to a rat (predator) exposure area. Familiar substrate is provided to permit burying, and mouse subjects are habituated to the apparatus prior to exposure to an amphetamine-activated rat. In comparison to toy-rat-exposed controls, rat-exposed BALB/c mice showed significantly more risk assessment [stretch attend posture (SAP) and stretch approach], freezing, and avoidance (time in the home chamber), and less time in contact with the wire mesh screen between itself and the threat stimulus. When BALB/c, C57BL/6, CD-1, and Swiss-Webster mice were compared in this test, the two inbred strains (BALB/c and C57BL/6) tended to show more extreme values of particular defensive behaviors, compared to the two outbred strains (Swiss-Webster and CD-1). C57BL/6 mice showed more avoidance and higher levels of SAP, freezing, and burying than BALB/c and more than one or both outbred strains as well. BALB/c mice showed little defensive burying, both in comparison to toy-exposed controls (Experiment 1), and in comparison to the three other strains in Experiment 2. These findings are somewhat at variance with characterizations of anxiety in C57BL/6 and BALB/c mice, based on tests utilizing novel areas and noxious stimuli, suggesting strain differences in defensiveness to such stimuli, compared to antipredator defense levels. Nonetheless, with the exception of burying in BALB/c mice, all strains showed all defensive behaviors measured to the rat stimulus. In particular, SAP levels were substantial in all strains tested, suggesting the usefulness of this test in assessment of the role of risk assessment in defense. PMID- 15135019 TI - A comparison of the sniff magnitude test and the University of Pennsylvania Smell Identification Test in children and nonnative English speakers. AB - The sniff magnitude test (SMT) is a reliable and rapid clinical test of olfactory function that is minimally dependent on cognitive and linguistic abilities. In this study, we compared performance on the SMT and University of Pennsylvania Smell Identification Test (UPSIT) in samples of children and nonnative English speakers. Previous research has shown that these populations perform poorly on the UPSIT as compared with young, healthy U.S. adults. Such performance differences may reflect variations in memory/cognition and language/culture rather than olfactory abilities. The UPSIT scores of children and of Indian and Chinese graduate students were found to be lower than those of young U.S. adults. By contrast, these groups did not perform more poorly than U.S. adults did on the SMT. The results are consistent with findings from our studies, with the elderly showing that performance on the UPSIT, but not the SMT, is significantly correlated with measures of memory, language and other cognitive abilities. The findings highlight the utility of the SMT when evaluating the olfactory ability of the very young, older adults and people with diverse linguistic and cultural backgrounds. PMID- 15135020 TI - Chronic infusion of the amylin antagonist AC 187 increases feeding in Zucker fa/fa rats but not in lean controls. AB - Numerous studies have established the pancreatic B-cell hormone amylin as an important anorectic peptide affecting meal-ending satiety. In the present study, we investigated the effect of a chronic infusion of the amylin antagonist AC 187 on food intake. The studies were performed using obese Zucker fa/fa rats, which are hyperamylinemic but have a defective leptin and insulin signaling system. A chronic intraperitoneal infusion of the amylin antagonist AC 187 (10 microg/kg/h) significantly increased dark phase and total food intake in Zucker but not in lean control rats. During the 8-day infusion experiment, AC 187 had no clear effect on body weight gain in either group. After acute administration, amylin and its agonist salmon calcitonin (sCT) equally reduced food intake in Zucker and lean control rats while cholecystokinin's (CCK) anorectic effect was weaker in the Zucker rats. We provide evidence for amylin being a potential long-term regulator of food intake because AC 187 increased food intake in obese fa/fa rats but not in lean control animals, which have low baseline amylin levels. Amylin may play some role as lipostatic feedback signal similar to leptin and insulin at least when the leptin and insulin feedback signaling systems are deficient. Despite basal hyperamylinemia in the Zucker rats, they do not seem to be less sensitive to the anorectic effects of amylin or its agonist sCT than respective controls. This contrasts with CCK whose anorectic action is reduced in Zucker rats when compared with lean controls. PMID- 15135021 TI - Effects of neonatal handling on the behavior and prolactin stress response in male and female rats at various ages and estrous cycle phases of females. AB - Neonatal handling induces behavioral and hormonal changes, characterized by reduced fear in novel environments, and lesser elevation and faster return to basal levels of plasma corticosterone, prolactin and adrenaline, in response to stressors in adulthood. The present study aimed to analyze the effects of neonatal handling from Days 1 to 10 postnatal on prolactin response to ether stress in male and female rats at three life periods: neonatal, peripubertal and adulthood. Moreover, adult females were tested in two different phases of the estrous cycle, i.e., diestrus and estrus. In another set of experiments, the behavior of peripubertal and adult males and females in estrus and diestrus was analyzed in the elevated plus maze test. Pups were either handled for 1 min (handled group) or left undisturbed (nonhandled group) during the first 10 days after delivery. In adults, in the handled females in diestrus, stress induced a lesser increase in plasma prolactin compared with nonhandled ones, as in males. However, in estrus, handled females showed no difference in the prolactin response to stress. In the elevated plus maze, handled females in diestrus, but not in estrus, showed higher locomotor activity compared with nonhandled ones. Peripubertal male and female rats handled during the neonatal period showed no difference in behavior in the elevated plus maze compared with nonhandled animals. Early-life stimulation can induce long-lasting behavioral and stress related hormonal changes, but they are not stable throughout life and phases of the estrous cycle. PMID- 15135022 TI - Preabsorptive factors are not the main determinants of intake depression induced by a high-protein diet in the rat. AB - The factors involved in the depression of food intake produced by a high-protein diet are still poorly understood. The aim of this study was to assess the role of several preingestive or preabsorptive factors likely to influence food intake when rats were fed ad libitum. Food intake was measured after modifying the composition of the high-protein diet, i.e., the type of proteins, or carbohydrates. Moreover, correlations between high-protein diet intake and the quantity of fluid ingested or stomach volume were studied. By varying the carbohydrate composition (sucrose/cornstarch) and the protein source (soy or gluten or total milk protein) of high-protein diets, we modified the orosensory properties of these diets. However, no differences in food intake were observed between these groups of rats during the transition phase or after adaptation, except during the first day of soy- or gluten-based diets when the depression of food intake was intensified. The depression of high-protein diet intake was neither the consequence of any delay necessary to increase the fluid intake induced by eating a high-protein diet nor due to a marked increase in stomach volume, which might explain enhanced satiety and decreased food intake through the activation of vagal afferent fibers. Our experiments do not indicate a preponderant role for oropharyngeal or preabsorptive factors in the depression of food intake induced by a high-protein diet. PMID- 15135023 TI - Relating ionisation of calcium chloride in saliva to bitterness perception. AB - Saliva plays a role in the perception of bitter, sour and salty tastes that are presumed to be derived from the concentration of free cations or anions ions dissolved in saliva. The role of ionisation of calcium in bitter taste was studied by determining binding in vitro mixture of saliva and protein solutions and in spit. In vitro, the addition of whey to calcium chloride solutions increased the calcium binding, pH and viscosity. The addition of saliva to these mixtures, the increased calcium binding and the induced small changes in viscosity and pH were thought not to contribute significantly to bitterness perception. Nonstimulated saliva, at pH 7.5, contained about 5 mM calcium, of which about one third was ionised. The bitter threshold of fully ionised calcium chloride in water varied between 1 and 15 mM among individuals. In spit, after tasting whey, ionised calcium was found to have increased at low, but decreased at high, calcium concentrations and varied 30% among individuals. Bitterness was related, on average, to the concentration of ionised calcium and not to the total concentration of calcium in spit. A general explicative model based on the composition of bulk saliva is discussed in relation to perception threshold and the likely importance of saliva from von Ebner's gland. PMID- 15135024 TI - Food hedonics and reinforcement as determinants of laboratory food intake in smokers. AB - Both the hedonic ratings and the reinforcing value of food have been considered to be determinants of food intake. The objective of this study was to compare the pleasurable ratings and the reinforcing value of food as determinants of energy intake. Seventy-four smokers were studied in food consumption and reinforcing value of food tasks prior to enrolling in a smoking-cessation treatment program. For the food consumption task, the participants tasted and consumed food ad lib from eight snack foods. The reinforcing value of the food task assessed how hard subjects would work for a preferred snack food. Results showed that food reinforcement was related to laboratory food intake, with those high in food reinforcement consuming significantly more calories (+114.4 kcal, P<.01) than did the participants low in food reinforcement. Food reinforcement was related to food intake for the preferred food as well as to total energy intake. Hedonics for the preferred food was related to food reinforcement but not to either measure of laboratory energy intake. In multiple-regression models, food reinforcement and the interaction of food reinforcement by sex were significant predictors of energy intake for the preferred food and for total energy intake, along with baseline hunger. In conclusion, energy intake in smokers in a laboratory setting is more strongly related to food reinforcement than to the hedonic ratings of food. PMID- 15135025 TI - Effects of neonatal oxytocin manipulations on male reproductive potential in prairie voles. AB - Oxytocin (OT) modulates adult mammalian sexual behavior, sperm production and transport, and steroidogenesis; however, the consequences of developmental manipulations of oxytocin have received little attention. The purpose of this experiment was to determine whether neonatal exposure to OT, an oxytocin antagonist (OTA), saline (SAL), or handling (HAN)-only would have long-term effects on reproductive potential in male prairie voles (Microtus ochrogaster). Adult males were observed for 24 h with a sexually receptive female and sexual behavior was recorded. Females were subsequently lavaged and smears were examined for sperm. Reproductive parameters including motility of epididymal sperm, testis weight, and plasma androgen levels were in the normal range. OT-treated males that did not mate within the first 30 min did not mate at all, and in comparison to controls, a higher proportion of those OT-treated and OTA-treated males that did mate did not transfer sperm to the females. OTA-treated males also had significantly higher testicular sperm concentrations than HAN-only males, and significantly lower epididymal sperm concentrations. These differences suggest that in males, developmental manipulations of OT may have the potential to influence the subsequent expression of sexual behavior and sperm transport. PMID- 15135026 TI - Behavioral and neurochemical effects induced by subchronic combined exposure to toluene at 40 ppm and noise at 80 dB-A in rats. AB - We investigated whether exposure to noise, in addition to its well-known potentiating effect on toluene-induced ototoxicity, may also exacerbate behavioral disturbances and brain neurochemical alterations produced by subchronic exposure to low toluene concentration. To test this hypothesis, we evaluated whether subchronic combined exposure (16 weeks, 104 h per week) to noise at 80 dB-A and toluene at 40 ppm potentiates the recently reported neurotoxic effects of subchronic exposure to 40 ppm toluene. Locomotor and rearing activities, sensitization to narcosis induced by acute toluene at high concentration, and tyrosine and tryptophan hydroxylase activities in the caudate putamen and hippocampus were investigated in both male and female rats. Our results confirm that subchronic exposure to 40 ppm toluene significantly decreases rearing activity and leads to a sensitization to toluene-induced narcosis, as evaluated by loss of righting reflex, but fails to demonstrate any adverse effect of noise, alone or in combination with toluene. Given that toluene has addictive properties, the lack of potentiating behavioral and neurochemical effect of noise is discussed with regards to a recent study that has shown that methamphetamine neurotoxicity is potentiated by exposure to loud noise. PMID- 15135027 TI - Norepinephrine turnover in brown and white adipose tissue after partial lipectomy. AB - Total body fat is restored after the surgical removal (i.e., partial lipectomy) of white adipose tissue (WAT), and this is accomplished via increases in the mass of nonexcised WAT pads. The underlying mechanism for this apparent regulation of total body fat is unknown. One possibility is via the sympathetic nervous system (SNS) innervation of WAT and brown adipose tissue (BAT) through the regulation of lipolysis and thermogenesis, respectively. Specifically, decreases in SNS activity might fuel lipectomy-induced body fat compensation through energy saved from decreased BAT thermogenesis and would promote lipid accretion through decreased WAT basal lipolysis. Therefore, we tested whether lipectomy triggered decreases in the SNS drive [as indicated by the norepinephrine turnover (NETO)] to nonexcised WAT or to BAT, at times before the lipectomy-induced fat pad mass compensation was complete. Siberian hamsters received either sham or bilateral epididymal WAT lipectomy, and NETO was measured in the remaining WAT and interscapular BAT (IBAT) before, and 3 and 6 weeks after surgery. Total dissected WAT, and inguinal and retroperitoneal WAT masses were significantly increased following lipectomy, whereas dorsal subcutaneous WAT and IBAT masses, as well as food intake, were unchanged. The only significant change in NETO was a marked decrease (approximately 90%) in IBAT NETO at Week 3 postlipectomy compared with the sham-lipectomized controls. These findings suggest that the lipid accretion of nonexcised WAT pads triggered by lipectomy may be partially fueled by decreased BAT thermogenesis, inasmuch as decreased IBAT NETO reflects decreased BAT heat production. PMID- 15135028 TI - Effects of stochasticity in models of the cell cycle: from quantized cycle times to noise-induced oscillations. AB - Noise and fluctuations are ubiquitous in living systems. Still, the interaction between complex biochemical regulatory systems and the inherent fluctuations ('noise') is only poorly understood. As a paradigmatic example, we study the implications of noise on a recently proposed model of the eukaryotic cell cycle, representing a complex network of interactions between several genes and proteins. The purpose of this work is twofold: First, we show that the inclusion of noise into the description of the system accounts for several recent experimental findings, as e.g. the existence of quantized cycle times in wee1- cdc25delta double-mutant cells of fission yeast. In the main part, we then focus on more general aspects of the interplay between noise and the dynamics of the system. In particular, we demonstrate that a stochastic description leads to qualitative changes in the dynamics, such as the emergence of noise-induced oscillations. These findings will be discussed in the light of an ongoing debate on models of cell division as limit-cycle oscillators versus checkpoint mechanisms. PMID- 15135029 TI - Strong altruism can evolve in randomly formed groups. AB - Although the conditions under which altruistic behaviors evolve continue to be vigorously debated, there is general agreement that altruistic traits involving an absolute cost to altruists (strong altruism) cannot evolve when populations are structured with randomly formed groups. This conclusion implies that the evolution of such traits depends upon special environmental conditions or additional organismic capabilities that enable altruists to interact with each other more than would be expected with random grouping. Here we show, using both analytic and simulation results, that the positive assortment necessary for strong altruism to evolve does not require these additional mechanisms, but merely that randomly formed groups exist for more than one generation. Conditions favoring the selection of altruists, which are absent when random groups initially form, can naturally arise even after a single generation within groups and even as the proportion of altruists simultaneously decreases. The gains made by altruists in a second generation within groups can more than compensate for the losses suffered in the first and in this way altruism can ratchet up to high levels. This is true even if altruism is initially rare, migration between groups allowed, homogeneous altruist groups prohibited, population growth restricted, or kin selection precluded. Until now random group formation models have neglected the significance of multigenerational groups-even though such groups are a central feature of classic "haystack" models of the evolution of altruism. We also explore the important role that stochasticity (effectively absent in the original infinite models) plays in the evolution of altruism. The fact that strong altruism can increase when groups are periodically and randomly formed suggests that altruism may evolve more readily and in simpler organisms than is generally appreciated. PMID- 15135030 TI - Noise-reduction through interaction in gene expression and biochemical reaction processes. AB - We demonstrate that interaction in gene expression and biochemical reaction processes has a significant influence on reducing fluctuations. Especially, we have found that the interaction between synthesized proteins and background molecules can reduce the fluctuation level in gene expression, which is a counter example to the intuition that background factors disturb information processing in genetic networks by increasing the noise level. This fact also indicates that the macromolecular crowding observed in actual cells can contribute to reduce the noise level. In addition, the noise-reduction phenomenon is not limited to the interaction between the proteins and background molecules, but can be applied to other reactions such as a dimerization process and the coupling of reactions with large fluctuations by intrinsic noise. Finally, on the basis of these results, we propose a new and plausible method for reducing the fluctuations generated in synthesized genetic networks, and also discuss the applicability of this method to the stabilization of system dynamics by using a toggle switch model. PMID- 15135031 TI - Thermodynamic constraints for biochemical networks. AB - The constraint-based approach to analysis of biochemical systems has emerged as a useful tool for rational metabolic engineering. Flux balance analysis (FBA) is based on the constraint of mass conservation; energy balance analysis (EBA) is based on non-equilibrium thermodynamics. The power of these approaches lies in the fact that the constraints are based on physical laws, and do not make use of unknown parameters. Here, we show that the network structure (i.e. the stoichiometric matrix) alone provides a system of constraints on the fluxes in a biochemical network which are feasible according to both mass balance and the laws of thermodynamics. A realistic example shows that these constraints can be sufficient for deriving unambiguous, biologically meaningful results. The thermodynamic constraints are obtained by comparing of the sign pattern of the flux vector to the sign patterns of the cycles of the internal cycle space via connection between stoichiometric network theory (SNT) and the mathematical theory of oriented matroids. PMID- 15135032 TI - New insights into vascular collapse and growth dynamics in solid tumors. AB - The experimentally-observed phenomenon of vascular collapse in tumors represents a significant barrier to the delivery of blood-borne therapeutic drugs, and has been attributed to the elevated tissue stresses resulting from confined proliferation of tumor cells. This paper presents a mathematical framework which describes the evolution of growth-induced stresses in tumors and gives new insights into both vascular collapse and tumor growth dynamics. The linear elastic description of anisotropic growth adopted here provides the mechanical model with a realistic constitutive basis, incorporating both the solid and stress-relaxation characteristics of soft biological tissues. A particular distribution of spatially non-uniform growth is proposed which is considered representative of a vascular tumor. The stress distribution associated with this growth pattern predicts the onset of vascular collapse, producing the well defined regions observed in vascular collapse experiments: a peripheral layer with open blood vessels adjacent to a region of vascular collapse, enclosing an inner region where the vessels are open. The model also highlights the roles of various tissue properties in inducing vascular collapse. Moreover, the tumor growth rates predicted by this model reflect experimental observations, with exponential growth taking place immediately following vascularization, followed by a period of exponential retardation. PMID- 15135033 TI - Space-irrelevant scaling law for fish school sizes. AB - Universal scaling in the power-law size distribution of pelagic fish schools is established. The power-law exponent of size distributions is extracted through the data collapse. The distribution depends on the school size only through the ratio of the size to the expected size of the schools an arbitrary individual engages in. This expected size is linear in the ratio of the spatial population density of fish to the breakup rate of school. By means of extensive numerical simulations, it is verified that the law is completely independent of the dimension of the space in which the fish move. Besides the scaling analysis on school size distributions, the integrity of schools over extended periods of time is discussed. PMID- 15135034 TI - The elusive action of sex-determining genes: mitochondria to the rescue? AB - According to the accepted dogma of mammalian sex determination, the Y-linked gene SRY initiates male development by inducing hitherto uncommitted somatic cells of the fetal gonad to develop into Sertoli cells. However, it has become evident that the correct functioning of an increasing number of genes on other chromosomes is required for testicular organogenesis. They include the SRY related gene, SOX9, which plays important roles in both sex determination and chondrogenesis, as well as genes responsible for the production of growth factors, i.e. fibroblast growth factor 9, platelet derived growth factor A, and the members of the insulin-receptor family of genes. It is known, moreover, that differences between the sexes begin to develop long before the differentiation of Sertoli cells, including an increase in gonadal size and cell proliferation, and accelerated development of XY embryos at early pre-implantation stages. There is also evidence of transcription of Y-linked, and of X-linked, genes and of an enhanced metabolic rate in XY embryos. Furthermore, the condition of true hermaphroditism does not fit into a simple genotype/phenotype relationship. The proposal that "testis-determining" genes act by increasing metabolic rates rather than directly determining Sertoli cell differentiation can account for a number of observations that do not fit the current model, including pregonadal sex differences, the activity of the same gene in different organ systems, and the frequent co-existence of sexual and somatic abnormalities. It also sheds light on the pervasive differences between metabolic rates of mammalian males and females, while the facts of true hermaphroditism can be viewed as remnants of temperature dependent sex determination in ectothermic vertebrates. Growing interest in mitochondria, which play a central role in the provision of energy to eukaryotic cells, makes a shift of paradigm from gonadal histology to energy metabolism timely, particularly since new techniques have become available for testing the hypothesis, and for widening the experimental approach to sex determination. If the hypothesis is correct, it would mean that male sex is determined by nuclear genes inherited from the father regulating the activity of maternally derived mitochondria. PMID- 15135035 TI - Advantageous indirect interactions in systems of competition. AB - Ecological foodwebs display complex networks of species interactions. Here we discuss how two species, whether directly interacting or not, can crucially affect each other 'indirectly' through their mutual associations with intermediary species. A technique is presented for quantifying these "indirect effects", so that a simple measure emerges for the degree of overall harm or advantage that a particular species encounters from another in the context of a given foodweb. If the system is one of pure competition, the "direct" interaction between any two species is of course harmful to both. But when the method is applied to such systems it predicts that, if all interactions are allowed for including the indirect, in a large proportion of pair interactions at least one species will be deriving benefit from the other. Computer-generated samples confirm that this proportion is accurately predicted. The many pathways for indirect interaction can thus often reverse the direct connection, so that competing species achieve an interaction here called 'Advantageous in A Community Context' (ACC). PMID- 15135036 TI - The parental investment conflict in continuous time: St. Peter's fish as an example. AB - The parental investment conflict considers the question of how much each sex should invest in each brood, thereby characterizing different animal species. Each species usually adopts a certain parental care pattern: female-care only, male-care only, biparental care, or even no parental care at all. The differences in care patterns are usually explained by the different costs and benefits arising from caring for the offspring in each animal species. This paper proposes a game-theoretical model to the parental investment conflict based on the parental behavior of St. Peter's fish. St. Peter's fish exhibit different parental care patterns, allowing the examination of the factors which determine the particular behavior in each mating. We present a continuous time, two-stage, asymmetric game, with two types of players: male and female. According to the model's results, three parental care patterns: male-only care, female-only care and biparental care, are possible evolutionarily stable strategies. The evolutionarily stable parental care pattern in a certain mating depends on a parent's increase in mortality due to parental care, and on its advantage from biparental care. These results may explain the different parental care patterns observed in a variety of animal species, including those found in the St. Peter's fish. PMID- 15135037 TI - Amino acid biogenesis, evolution of the genetic code and aminoacyl-tRNA synthetases. AB - The aminoacyl-tRNA synthetases (aaRSs) ensure the fidelity of the translation of the genetic code, covalently attaching appropriate amino acids to the corresponding nucleic acid adaptor molecules-tRNA. The fundamental role of aminoacylation reaction catalysed by aaRSs implies that representatives of the family are thought to be among the earliest proteins to appear. Based on sequence analysis and catalytic domain structure, aaRSs have been partitioned into two classes of 10 enzymes each. However, based on the structural and sequence data only, it will not be easily understood that the present partitioning is not governed by chance. Our findings suggest that organization of amino acid biosynthetic pathways and clustering of aaRSs into different classes are intimately related to one another. A plausible explanation for such a relationship is dictated by early link between aaRSs and amino acids biosynthetic proteins. The aaRSs catalytic cores are highly relevant to the ancient metabolic reactions, namely, amino acids and cofactors biosynthesis. In particular we show that class II aaRSs mostly associated with the primordial amino acids, while class I aaRSs are usually related to amino acids evolved lately. Reasoning from this we propose a possible chronology of genetic code evolution. PMID- 15135038 TI - Dynamics of T cell activation threshold tuning. AB - T lymphocytes are believed to alter their sensitivity to TCR stimulation by means of a tunable cellular activation threshold. We present two modelling examples which show that the concept of a tunable threshold can be made mechanistically plausible. The tunable threshold is treated as an emergent property of the dynamics of the T cell's signalling machinery. In addition, we discuss how the dynamic properties of activation threshold tuning can be determined experimentally with the aid of these two models. We propose a novel 'avidity selection' mechanism for the initial stages of the immune response, based on the properties of the T cell activation threshold tuning mechanism we propose for the commitment to differentiation. Our main finding is that activation threshold tuning allows T cells to respond to relevant ligands with a detection threshold that is (i) uniform across both the T cell repertoire and the secondary lymphoid tissues, while (ii) retaining tolerance to autostimulation. Our analysis indicates that central tolerance enhances the efficiency of peripheral tolerance, casting new light on the role of negative selection in the thymus. PMID- 15135039 TI - Phenological resonance and quantum life history. AB - The principle of 'quantum life history' is proposed here as a complementary viewpoint to current modeling of body size and life history evolution which usually considers a 'fast-slow continuum' of covarying life history traits. This principle emphasizes the discrete (and primary) nature of development time caused by the effect of phenological resonance (the compliance of development time with periodicities of earth rotation). The body mass, in turn, complies with development time, which generates body mass attractors. This principle is illustrated with mammals as exemplary group. The adaptive radiation of Cenozoic mammals is supposed to proceed as a competition-driven diversification of body sizes and development times around the strongest (year-long) resonant mode of development time corresponding to body mass of about 1 kg. Mammals with this body mass are shown here to have a largest genome size and a lowest (body mass corrected) basal metabolic rate. This extends the previously reported negative relation between genome size and metabolic rate to the realm of nonlinearity, and suggests that selection against the accumulation of non-coding DNA in the genome is relaxed in mammals with this body mass. PMID- 15135040 TI - Scalar modeling and analysis of a 3D biochemical reaction model. AB - For many systems it is advantageous if analysis and modeling can be accomplished from a scalar time series because this greatly facilitates the experimental setup. Moreover, in real-life systems it is hardly true that all the state variables are available for analysis and modeling. Since the late 1980s, techniques have been put forward for building mathematical models from a scalar time series. One of the objectives of this paper is to verify if it is possible to obtain global non-linear models (non-linear differential equations) from scalar time series. Such data are obtained using a model of biochemical reaction with aperiodic (chaotic) oscillations as recently observed in the case of a glycolytic reaction (Nielsen, K., Sorensen, P.G., Hynne, F., 1997. Chaos in glycolysis. J Theor. Biol. 186, 303-306.). The main objective, however, is to investigate which state variable is more convenient for the task in practice. It is shown that observability indices seem to quantify quite well which variable should be preferred as the observable. The validity of the results are established performing rigorous topological analysis on the original system and the obtained models. The influence of noise, always present in experimental time series, on the dynamics underlying such a system is also investigated. PMID- 15135041 TI - Obligate vertebrate scavengers must be large soaring fliers. AB - Among extant vertebrates, only the 23 species of vulture are obligate scavengers. We use an energetic modelling approach to explore the constraints imposed by an obligate scavenging lifestyle, and to ask whether obligate scavengers must always be avian and generally large-bodied users of soaring flight. Our model found that aerial scavengers always out-competed postulated terrestrial ones, mainly because flight allows area to be searched much more rapidly for carrion. Soaring was favoured over flapping flight because the reduction in flight speed (and so rate of area search) was more than compensated for by the decrease in the costs of transport. Large individual size is selected for if carrion is available in large packages, when obligate scavenger feed only infrequently, and so must be able to survive on body reserves in the periods between discovering food falls. In the absence of avian radiation, an obligate terrestrial scavenger seems energetically feasible, but we argue that such a beast is unlikely to have evolved. In birds, in order to become exclusive scavengers, vultures have needed to specialize for efficient soaring flight as a low energy form of travel, and as a consequence they have lost the agility needed to kill prey. In mammals, however, no comparable trade-off occurs. So for terrestrial carnivores there is probably no strong selection pressure towards being an exclusive scavenger. Indeed it will perhaps always be more advantageous to retain the flexibility of obtaining food by either predation or scavenging. PMID- 15135042 TI - alpha-Synuclein regulation of the dopaminergic transporter: a possible role in the pathogenesis of Parkinson's disease. AB - Parkinson's disease (PD) is a slow progressive neurodegenerative disorder. Recent evidence suggests a central role for alpha-synuclein, a protein of unknown function, in the genesis of PD. The phenomenon of selective degeneration of dopaminergic neurons in PD may be linked to the potential toxicity of dopamine itself and aberrations in the processes which regulate dopamine content may underlie the pathogenesis of this disease. Here, we review a vital role of alpha synuclein in the modulation of dopamine transporter (DAT) function, and describe how disruption of this modulatory process permits increased re-uptake of high levels of intracellular dopamine by DAT, causing profound neurotoxicity. PMID- 15135043 TI - Human limbal epithelium contains side population cells expressing the ATP-binding cassette transporter ABCG2. AB - Many types of organ-specific stem cells have been recently shown to exhibit a side population (SP) phenotype based on their ability to efflux Hoechst 33342 dye. Because stem cells from corneal epithelium reside in the basal layer of the limbal epithelium, the purpose of this study was to examine whether the limbal epithelium contains SP cells. The ATP-binding cassette transporter Bcrp1/ABCG2 is reported to contribute to the SP phenotype in cells from several diverse sources. Here we show data from fluorescence-activated cell sorting and real-time quantitative RT-PCR analysis showing that harvested limbal epithelial cells contain SP cells expressing ABCG2. Immunofluorescence revealed that a portion of limbal epithelial basal cells expressed ABCG2. Data indicate that ABCG2 positive limbal epithelial cells are putative corneal epithelial stem cells. PMID- 15135044 TI - Molecular and functional analysis of Caenorhabditis elegans CHIP, a homologue of Mammalian CHIP. AB - A recently identified molecule C-terminus of Hsc70 interacting protein (CHIP) has been reported to be an E3 ubiquitin ligase collaborating with molecular chaperones for the degradation of misfolded or unfolded proteins. The physiological roles of CHIP in animal and plant development remain largely unknown. Here, we show that the knockdown of CeCHIP by RNAi and knockout by a deletion mutation arrests the development of the animal at the larval stage. CeCHIP expresses ubiquitously in all tissues but there are tissue specific variations of expression level. CeCHIP produces dose dependent phenotypes in vivo. Over expression of CHIP causes embryonic lethality, while a comparatively lower level of over expression causes locomotion and egg laying defects, and the CHIP over expressed animals form dauers at a higher temperature. PMID- 15135045 TI - Mapping of the insulin-like growth factor II binding site of the Type I insulin like growth factor receptor by alanine scanning mutagenesis. AB - The Type I insulin-like growth factor receptor is a physiological receptor for insulin-like growth factor II (IGF-II). To characterize the molecular basis of the receptor's ligand binding properties, we have examined the effects of alanine mutations of residues in the ligand binding site of the receptor on its affinity for IGF-II. The functional epitope for IGF-II comprises residues in the N terminal L1 domain and residues at the C-terminus of the alpha subunit. Cysteine rich domain residues do not appear to be critical for IGF-II binding. PMID- 15135046 TI - Low-density lipoprotein receptor-related protein interacts with MafB, a regulator of hindbrain development. AB - The intracellular domain (ICD) of the low-density lipoprotein receptor-related protein (LRP) functionally interacts with adaptor proteins both as an integral part of the receptor polypeptide and after proteolytic release. Identification of such adaptors has been difficult because the ICD is self-activating in conventional transcription factor-based yeast two-hybrid screens. We adopted an alternative screen for the ICD that depends on the activation of the Ras signaling pathway and uncovered the transcription factor MafB as novel ICD interacting protein. MafB is a regulator of hindbrain segmentation and interacts with the ICD through a leucine zipper domain. The ICD co-localizes with MafB to the nucleus and negatively regulates its transcriptional activity, suggesting a possible role for LRP in brain development. PMID- 15135047 TI - Hsp25, a member of the Hsp30 family, promotes inclusion formation in response to stress. AB - Protein aggregates are oligomeric complexes of misfolded proteins, and serve as the seeds of inclusion bodies termed aggresomes in the cells. Heat shock proteins (Hsps) prevent misfolding and aggregate formation. Here, we found that only avian Hsp25 dominantly accumulated in the aggresomes induced by proteasome inhibition. Molecular cloning of chicken Hsp25 (cHsp25) revealed that it belongs to the Hsp30 family, which is a subfamily of the alpha-crystallin/small Hsp gene family. Unexpectedly, overexpression of cHsp25 into HeLa cells promoted inclusion formation whereas overexpression of mouse Hsp27 and its chicken homologue did not. These results suggest that cHsp25 acts differently from other small Hsps on protein aggregates. PMID- 15135048 TI - SH3P2 in complex with Cbl and Src. AB - In this report, we describe SH3P2, an SH3-domain containing protein, as a novel Cbl-interacting molecule that is a substrate of tyrosine kinase Src. We identified a specific polyproline motif of Cbl responsible for binding of SH3P2 and Src, and observed mutual sequestration of Src and SH3P2 from monomer Cbl molecules. In adherent cells, SH3P2 associated with Cbl and fibrilar actin and was localized at focal contacts in fibroblasts as well as at the apical part of podosome rings in differentiated osteoclasts. Our data implicate that SH3P2, a novel component of adhesion sites, is involved in Cbl and Src-mediated pathways. PMID- 15135049 TI - Recombinant ATPases of the yeast 26S proteasome activate protein degradation by the 20S proteasome. AB - The 26S proteasome contains a proteolytic core, 20S proteasome, and its regulatory particle, 19S complex. That regulatory particle contains six ATPases that are involved in unfolding and translocation of substrates to the 20S proteasome's catalytic chamber. We expressed ATPase-encoding genes of the regulatory particle of Saccharomyces cerevisiae and found that some recombinant ATPases can self-assemble into a high-molecular-weight protein complex in Escherichia coli. Purification of the Rpt1Rpt2 hetero-complex and the Rpt4 homo complex for functional characterization demonstrated their contribution to energy dependent protein degradation. Our finding, production of a functional subunit of the 19S regulatory particle in bacteria, is a simpler and technically advanced system to functionally characterize individual subunits. PMID- 15135050 TI - Suppression of a phosphatidylinositol 3-kinase signal by a specific spliced variant of Drosophila PTEN. AB - Drosophila PTEN (dPTEN) plays indispensable roles in the development of Drosophila melanogaster by controlling cell size and number. Although three potential spliced forms of dPTEN have been isolated, functional distinction among these forms remains elusive. In this study, we demonstrate that all spliced forms of dPTEN dephosphorylate phosphatidylinositol 3,4,5-trisphosphate (PI(3,4,5)P(3)); however, PI(3,4,5)P(3)-dependent activation of Drosophila Akt is suppressed specifically by one of three spliced forms, dPTEN3. Further, dPTEN3 dramatically changes its expression during the Drosophila development, while the other forms are expressed throughout the development. Our results suggest that dPTEN3 is the predominant spliced form that participates in PI(3,4,5)P(3) mediated signaling pathways. PMID- 15135051 TI - Paracoccus pantotrophus NapC can reductively activate cytochrome cd1 nitrite reductase. AB - The oxidized "as isolated" form of Paracoccus pantotrophus cytochrome cd1 nitrite reductase has a bis-histidinyl coordinated c heme and a histidine/tyrosine coordinated d1 heme. This form of the enzyme has previously been shown to be kinetically incompetent. Upon reduction, the coordination of both hemes changes and the enzyme is kinetically activated. Here, we show that P. pantotrophus NapC, a tetraheme c-type cytochrome belonging to a large family of such proteins, is capable of reducing, and hence activating, "as isolated" cytochrome cd1. NapC is the first protein from P. pantotrophus identified as being capable of this activation step and, given the periplasmic co-location and co-expression of the two proteins, is a strong candidate to be a physiological activation partner. PMID- 15135052 TI - The effect of raft lipid depletion on microvilli formation in MDCK cells, visualized by atomic force microscopy. AB - We have investigated whether raft lipids of Madin-Darby canine kidney (MDCK) cells play any role in microvilli maintenance using a combination of atomic force microscopy (AFM) and laser scanning confocal microscopy. MDCK cells were treated to reduce the amount of sphingolipids, cholesterol, or both and subsequently imaged, in buffer solution, using AFM. It was observed that inhibition of either sphingolipid or cholesterol biosynthesis led to a reduction in the number of microvilli on the surface of MDCK cells. However, this effect was not uniform across the monolayer, with some cells resembling those in untreated controls. The subsequent extraction of cholesterol from cells grown in the presence of inhibitors led to a further reduction in microvilli on the surface of the cells and, in some cases, resulted in monolayers devoid of full length microvilli. Significantly, smaller spikes were observed on the surface of the smoother cells. PMID- 15135053 TI - X-ray structure of tRNA pseudouridine synthase TruD reveals an inserted domain with a novel fold. AB - Pseudouridine synthases catalyse the isomerisation of uridine to pseudouridine in structural RNA. The pseudouridine synthase TruD, that modifies U13 in tRNA, belongs to a recently identified and large family of pseudouridine synthases present in all kingdoms of life. We report here the crystal structure of Escherichia coli TruD at 2.0 A resolution. The structure reveals an overall V shaped molecule with an RNA-binding cleft formed between two domains: a catalytic domain and an insertion domain. The catalytic domain has a fold similar to that of the catalytic domains of previously characterised pseudouridine synthases, whereas the insertion domain displays a novel fold. PMID- 15135054 TI - The insect antimicrobial peptide, L-pyrrhocoricin, binds to and stimulates the ATPase activity of both wild-type and lidless DnaK. AB - Recent reports have indicated that insect antimicrobial peptides kill bacteria by inhibiting the molecular chaperone DnaK. It was proposed that the antimicrobial peptide, all-L-pyrrhocoricin (L-PYR), binds to two sites on DnaK, the conventional substrate-binding site and the multi-helical C-terminal lid, and that inhibition of DnaK comes about from the lid mode of binding. In this report, we show using two different assays that L-PYR binds to and stimulates the ATPase activity of both wild-type and a lidless variant of DnaK. Our study shows that L PYR interacts with DnaK much like the all-L NR (NRLLLTG) peptide, which is known to bind in the conventional substrate-binding site of DnaK. L-PYR antimicrobial activity is thus a consequence of the competitive inhibition of bacterial DnaK. PMID- 15135055 TI - Endogenous SHIP2 does not localize in lipid rafts in 3T3-L1 adipocytes. AB - SH2 domain containing inositol polyphosphate 5-phosphatase (SHIP2) dephosphorylates phosphatidylinositol 3,4,5-trisphosphate (PtdIns(3,4,5)P(3)) into phosphatidylinositol 3,4-bisphosphate (PtdIns(3,4)P(2)). SHIP2 knock-out mice demonstrated that SHIP2 acts as a negative regulator of insulin cascade in vivo. Our two-hybrid study showed that SHIP2 interacts with c-Cbl associated protein (CAP) and c-Cbl, implicated in the insulin signaling. As some proteins implicated in insulin signaling, like insulin receptor, CAP, c-Cbl or TC10, were reported to localize in lipid rafts, we addressed the same question for SHIP2. SHIP2 was detected in the non-raft fraction in CHO-IR, C2C12 myotubes and 3T3-L1 adipocytes except when it is overexpressed in CHO-IR, where we detected SHIP2 in the raft fraction. PMID- 15135056 TI - Cross talk between DevS sensor kinase homologue, Rv2027c, and DevR response regulator of Mycobacterium tuberculosis. AB - Rv2027c is a putative orphan histidine sensor kinase that bears strong homology to DevS of the hypoxia-responsive DevR-DevS two-component system in M. tuberculosis. The cytosolic C-terminal domain of Rv2027c protein (Rv2027c(194)) was overexpressed in E. coli and biochemically characterized. Rv2027c(194) underwent autophosphorylation at a conserved His(392) residue and engaged in phosphotransfer with DevR response regulator. The rates of autophosphorylation and the stabilities of the phosphorylated species were broadly similar in Rv2027c and DevS. However, unlike DevS, Rv2027c utilized Ca(2+) as an alternative divalent ion during autophosphorylation. In contrast to DevS which completed phosphotransfer to DevR in 5-10 min, phosphotransfer from Rv2027c approximately P was only partial at 30 min. Unlike devS transcription that was hypoxia responsive, Rv2027c transcript levels were not upregulated from basal levels during hypoxia. The differential regulation of devS and Rv2027c genes, the ability of Rv2027c to utilize Ca(2+) as a divalent cation in autophosphorylation at physiological concentrations and to engage in phosphotransfer with DevR suggests that the DevR regulon could be modulated by more than one environmental cue relayed through DevS and Rv2027c. PMID- 15135057 TI - Reconstitution and functional characterization of the unusual bi-subunit type I DNA topoisomerase from Leishmania donovani. AB - Leishmania donovani topoisomerase I is an unusual bi-subunit enzyme. The activity of the enzyme has been detected when the genes of the individual subunits were co expressed in yeast [J. Biol. Chem. 278 (2003) 3521]. Here, we report for the first time, the in vitro reconstitution of the two recombinant proteins, LdTOP1L and LdTOP1S, corresponding to the large and small subunits and localization of the active enzyme in both the nucleus and kinetoplast. The proteins were purified from bacterial extract and the activity was measured by plasmid DNA relaxation assay. LdTOP1L and LdTOP1S form a direct 1:1 heterodimer complex through protein protein interaction. Under standard relaxation assay condition (50 mM KCl and 10 mM Mg(2+)), reconstituted enzyme (LdTOP1LS) showed reduced processivity as well as 2-fold reduced affinity for DNA compared to eukaryotic monomeric rat liver topoisomerase I (RLTOP1). Cleavage assay at various salt concentrations reveals that Camptothecin (CPT) enhanced the formation of "cleavable complex" at low salt. Interaction between the two subunits leading to the formation of an active complex could be explored as an insight for development of new therapeutic agents with specific selectivity. PMID- 15135058 TI - Furin cleavage of the HIV-1 Tat protein. AB - Extracellular human immunodeficiency virus-1 (HIV-1) Tat protein and Tat-derived peptides are biologically active but mechanisms of Tat processing are not known. Within the highly conserved basic region of HIV-1 Tat protein (amino acids, a.a. 48-56), we identified two putative furin cleavage sites and showed that Tat protein was cleaved in vitro at the second site, RQRR? (a.a. 53-56?). This in vitro cleavage was blocked by a monoclonal antibody that binds near the cleavage site or by the furin inhibitor alpha-1 PDX. Monocytoid cells rich in furin also degraded Tat and this process was slowed by the furin inhibitor or the specific monoclonal antibody. Furin processing did not affect the rates for Tat uptake and nuclear accumulation in HeLa or Jurkat cells, but the transactivation activity was greatly reduced. Furin processing is a likely mechanism for inactivating extracellular HIV-1 Tat protein. PMID- 15135059 TI - Integrative analysis of multiple gene expression profiles applied to liver cancer study. AB - A statistical method for combining multiple microarray studies has been previously developed by the authors. Here, we present the application of the method to our hepatocellular carcinoma (HCC) data and report new findings on gene expression changes accompanying HCC. From the cross-verification result of our studies and that of published studies, we found that single microarray analysis might lead to false findings. To avoid those pitfalls of single-set analyses, we employed our effect size method to integrate multiple datasets. Of 9982 genes analyzed, 477 significant genes were identified with a false discovery rate of 10%. Gene ontology (GO) terms associated with these genes were explored to validate our method in the biological context with respect to HCC. Furthermore, it was demonstrated that the data integration process increases the sensitivity of analysis and allows small but consistent expression changes to be detected. These integration-driven discoveries contained meaningful and interesting genes not reported in previous expression profiling studies, such as growth hormone receptor, erythropoietin receptor, tissue factor pathway inhibitor-2, etc. Our findings support the use of meta-analysis for a variety of microarray data beyond the scope of this specific application. PMID- 15135060 TI - Identification of an Arabidopsis inorganic pyrophosphatase capable of being imported into chloroplasts. AB - An Arabidopsis cDNA coding for a previously uncharacterized isoform of inorganic pyrophosphatase was isolated. It was used to complement an E. coli mutant, demonstrating that it coded for an active enzyme. MgCl(2) was necessary for the protein's activity, whilst NaF inhibited it. The K(m) for pyrophosphate and the pH optimum of the protein was determined. The gene coding for this protein was expressed in all tissues, and its expression in rosette leaves was induced by incubation on metabolizable sugars. In vitro import experiments demonstrated that the protein could be imported into chloroplasts and localized to the stromal compartment. PMID- 15135061 TI - Rrs1p, a ribosomal protein L11-binding protein, is required for nuclear export of the 60S pre-ribosomal subunit in Saccharomyces cerevisiae. AB - Rrs1p is a ribosomal protein L11-binding protein in Saccharomyces cerevisiae. We have obtained temperature-sensitive rrs1 mutants by random PCR mutagenesis. [(3)H]Methionine pulse-chase analysis reveals that the rrs1 mutations cause a defect in maturation of 25S rRNA. Ribosomal protein L25-enhanced green fluorescent protein, a reporter of the 60S ribosomal subunit, concentrates in the nucleus with enrichment in the nucleolus when the rrs1 mutants are shifted to the restrictive temperature. These results suggest that Rrs1p stays on the pre-60S particle from the early stage to very late stage of the large-subunit maturation and is required for export of 60S subunits from the nucleolus to the cytoplasm. PMID- 15135062 TI - Identification of a novel protein 3a from severe acute respiratory syndrome coronavirus. AB - The open reading frame 3 of the severe acute respiratory syndrome coronavirus (SARS-CoV) genome encodes a predicted protein 3a, consisting of 274 amino acids, that lacks any significant similarities to any known protein. We generated specific antibodies against SARS protein 3a by using a synthetic peptide (P2) corresponding to amino acids 261-274 of the putative protein. Anti-P2 antibodies and the sera from SARS patients could specifically detect the recombinant SARS protein 3a expressed in Escherichia coli and in Vero E6 cells. Expression of SARS protein 3a was detected at 8-12 h after infection and reached a higher level after approximately 24 h in SARS-CoV-infected Vero E6 cells. Protein 3a was also detected in the alveolar lining pneumocytes and some intra-alveolar cells of a SARS-CoV-infected patient's lung specimen. Recombinant protein 3a expressed in Vero E6 cells and protein 3a in the SARS-CoV-infected cells was distributed over the cytoplasm in a fine punctate pattern with partly concentrated staining in the Golgi apparatus. Our study demonstrates that SARS-CoV indeed expresses a novel protein 3a, which is present only in SARS-CoV and not in other known CoVs. PMID- 15135063 TI - Cellular growth inhibition by TGF-beta1 involves IRS proteins. AB - In Mv1Lu cells, insulin partially reverses transforming growth factor-beta1 (TGF beta1) growth inhibition in the presence of alpha5beta1 integrin antagonists. TGF beta1 appears to induce phosphorylation of IRS-2 in these cells; this is inhibited by a TGF-beta antagonist known to reverse TGF-beta growth inhibition. Stable transfection of 32D myeloid cells (which lack endogenous IRS proteins and are insensitive to growth inhibition by TGF-beta1) with IRS-1 or IRS-2 cDNA confers sensitivity to growth inhibition by TGF-beta1; this IRS-mediated growth inhibition can be partially reversed by insulin in 32D cells stably expressing IRS-2 and the insulin receptor (IR). These results suggest that growth inhibition by TGF-beta1 involves IRS proteins. PMID- 15135065 TI - Evidence that human genes of modular proteins have retained significantly more ancestral introns than their fly or worm orthologues. AB - Comparison of the exon-intron structures of human, fly and worm orthologues of mosaic genes assembled from class 1-1 modules by exon-shuffling has revealed that human genes retained significantly more of the original inter-module introns than their protostome orthologues. It is suggested that the much higher rate of intron loss in the worm- and insect lineages than in the chordate lineage reflects their greater tendency for genome compaction. PMID- 15135064 TI - Bidirectional role of orphan nuclear receptor RORalpha in clock gene transcriptions demonstrated by a novel reporter assay system. AB - Circadian rhythms are generated by an extremely complicated transcription translation feedback loop. To precisely analyze the molecular mechanisms of the circadian clock, it is critical to monitor multiple gene expressions and/or interactions with their transcription factors simultaneously. We have developed a novel reporter assay system, the tricolor reporter in vitro assay system, which consists of green- and red-emitting Phrixothrix luciferases as dual reporters and blue-emitting Renilla luciferase as internal control. We have successfully employed this system in analyzing the effects of clock gene products on the enhancer elements of Per1 and Bmal1 promoters. The results indicate that the orphan nuclear receptor RORalpha regulates bidirectionally Bmal1 (positively) and Per1 (negatively) transcriptions simultaneously. PMID- 15135066 TI - Neuronal traits are required for glucose-induced insulin secretion. AB - The transcriptional repressor RE1 silencer transcription factor (REST) is an important factor that restricts some neuronal traits to neurons. Since these traits are also present in pancreatic beta-cells, we evaluated their role by generating a model of insulin-secreting cells that express REST. The presence of REST led to a decrease in expression of its known target genes, whereas insulin expression and its cellular content were conserved. As a consequence of REST expression, the capacity to secrete insulin in response to mitochondrial fuels, a particularity of mature beta-cells, was impaired. These data provide evidence that REST target genes are required for an appropriate glucose-induced insulin secretion. PMID- 15135067 TI - Transactivation of erbB2 by short and long isoforms of leptin receptors. AB - We generated kinase-positive and kinase-negative erbB2 tagged with YFP and the long form of leptin receptor (LEPRb) tagged with CFP. Both were as active as their untagged analogs. Both short and long isoforms of leptin receptor phosphorylated and thereby activated erbB2 upon leptin binding and enhanced MAPK activity. Our results unveil a novel route by which leptin may provoke erbB2's phosphorylation and thus enhance its oncogenic potential independently of HER family ligands or its overexpression. Using FRET technology in living cells, we found no evidence of complex formation between erbB2 and prolactin or leptin receptors, indicating that the transactivation occurs through an indirect interaction. PMID- 15135068 TI - Secreted Klotho protein in sera and CSF: implication for post-translational cleavage in release of Klotho protein from cell membrane. AB - Klotho mutant mice exhibit a set of phenotypes resembling human ageing. Although the function of Klotho remains unclear, mediation of its pleiotropic functions by putative humoral factor(s) has been presumed. Newly established antibodies against Klotho allowed the detection of secreted Klotho, a candidate for the putative humoral factor, in sera and cerebrospinal fluid. Surprisingly the secreted Klotho was 130 kDa, in contrast to the 70 kDa predicted form from klotho gene transcripts. The secreted as well as the membrane-bound Klotho proteins were suggested to form oligomerized complex. These results delineate post-translation processing of Klotho and possible regulatory mechanisms for secretion of Klotho in vivo. PMID- 15135069 TI - Regulation of the yeast phospholipid hydroperoxide glutathione peroxidase GPX2 by oxidative stress is mediated by Yap1 and Skn7. AB - The GPX2 gene encodes a homologue of phospholipid hydroperoxide glutathione peroxidase in Saccharomyces cerevisiae. The GPX2 promoter contains three elements the sequence of which is completely consistent with the optimal sequence for the Yap1 response element (YRE). Here, we identify the intrinsic YRE that functions in the oxidative stress response of GPX2. In addition, we discovered a cis-acting element (5'-GGCCGGC-3') within the GPX2 promoter proximal to the functional YRE that is necessary for H(2)O(2)-induced expression of GPX2. We present evidence showing that Skn7 is necessary for the oxidative stress response of GPX2 and is able to bind to this sequence. We determine the optimal sequence for Skn7 to regulate GPX2 under conditions of oxidative stress to be 5'-GGC(C/T)GGC-3', and we designate this sequence the oxidative stress-responsive Skn7 response element. PMID- 15135070 TI - Different stabilities and denaturation pathways for structurally related aromatic amino acid hydroxylases. AB - We have compared the urea stability of the human aromatic amino acid hydroxylases (AAAHs), key enzymes involved in neurotransmitter biosynthesis and amino acid homeostasis. Tyrosine-, tryptophan- and phenylalanine hydroxylase (TH, TPH and PAH, respectively) were transiently activated at low urea concentrations and rapidly inactivated in >3 M urea. The denaturation of TH occurred through two cooperative transitions, with denaturation midpoints of 1.41+/-0.06 and 5.13+/ 0.05 M urea, respectively. Partially denatured human TH (hTH) retained more of its secondary structure than human PAH (hPAH), and was found to exist as tetramers, whereas hPAH dissociated into dimers. Furthermore, the urea-induced aggregation of hPAH was 100-fold higher than for hTH. These results suggest that the denatured state properties of the AAAHs contribute significantly to the stability of these enzymes and their tolerance towards missense mutations. PMID- 15135071 TI - New insights on glycerol transport in Saccharomyces cerevisiae. AB - Previous studies evidenced in Saccharomyces cerevisiae the activity of a H(+)/glycerol symport, derepressed by growth on non-fermentable carbon sources, later associated with GUP1 and GUP2 genes. It was also demonstrated that only the combined deletion of GUP1, GUP2 together with GUT1 (glycerol kinase) abolished active transport in ethanol-induced cells. In this work, we show that a glycerol H(+)/symport, with identical characteristics to the previously described, was found in gup1gup2gut1 grown under salt-stress, particularly high in cells collected during diauxic-shift. These results suggest different roles for Gup1/2p than glycerol transport. The gene encoding for glycerol active uptake is thus yet unknown. PMID- 15135072 TI - The effect of phalloidin and jasplakinolide on the flexibility and thermal stability of actin filaments. AB - In this work the effect of phalloidin and jasplakinolide on the dynamic properties and thermal stability of actin filaments was studied. Temperature dependent fluorescence resonance energy transfer measurements showed that filaments of Ca-actin became more rigid in the presence of phalloidin or jasplakinolide. Differential scanning calorimetric data implied that the stiffer filaments also had greater thermal stability in the presence of phalloidin or jasplakinolide. The fluorescence and calorimetric measurements provided evidences that the extent of stabilization by jasplakinolide was greater than that by phalloidin. PMID- 15135073 TI - Glioblastoma cells block radiation-induced programmed cell death of endothelial cells. AB - We demonstrate that human umbilical vein endothelial cells (HUVEC) grown in co culture (CC) with U87 glioblastoma cells transfected with green fluorescent protein (GFP-U87) exhibit resistance to radiation-mediated apoptosis. cDNA macroarray analysis reveals increases in the accumulation of RNAs for HUVEC genes encoding cell adhesion molecules, growth factor-related proteins, and cell cycle regulatory/DNA repair proteins. An increase in protein expression of integrin alphav, integrin beta1, MAPK(p42), Rad51, DNA-PK(CS), and ataxia telangiectasia gene (ATM) was detected in HUVEC grown in CC with GFP-U87 cells compared with HUVEC grown in mono-culture. Treatment with anti-VEGF antibody decreases the expression of integrin alphav, integrin beta1, DNA-PK(CS) and ATM with a corresponding increase in ionizing radiation (IR)-induced apoptosis. These data support the concept that endothelial cells growing in the tumor microenvironment may develop resistance to cytotoxic therapies due to the up-regulation by tumor cells of endothelial cells genes associated with survival. PMID- 15135074 TI - Ion channel-like activity of the antimicrobial peptide tritrpticin in planar lipid bilayers. AB - The cationic peptide tritrpticin (VRRFPWWWPFLRR, Trp3) has a broad action spectrum, acting against Gram-positive and Gram-negative bacteria, as well as some fungi, while also displaying hemolytic activity. We have studied the behavior of Trp3 in planar lipid bilayers (or black lipid membrane - BLM) and were able to demonstrate its ion channel-like activity. Channel-like activity was observed in negatively charged azolectin BLM as a sudden appearance of discrete current fluctuations upon application of a constant voltage across the membrane. Trp3 formed large conductance channels (500-2000 pS) both at positive and negative potentials. In azolectin bilayers, the predominant ion-channel activity was characterized by very regular and discrete current steps (corresponding to openings) of uniform amplitude, which exhibited relatively long residence times (of the order of seconds). Occasionally, multiple conductance steps were observed, indicating the simultaneous presence of more than one open pore. In bilayers of zwitterionic diphytanoylphosphatidyl choline (DPhPC) Trp3 also showed ion-channel activity, but in a much less frequent and less prominent way. Studies of ion selectivity indicated that Trp3 forms a cation-selective channel. These results should contribute to the understanding of the molecular interactions and mechanism of action of Trp3 in lipid bilayers and biological membranes. PMID- 15135075 TI - Analysis of the S. pombe signalling scaffold protein Cdc11p reveals an essential role for the N-terminal domain in SIN signalling. AB - The initiation of cytokinesis in the fission yeast Schizosaccharomyces pombe is signalled by the septation initiation network (SIN). Signalling originates from the spindle pole body (SPB), where SIN proteins are anchored by a scaffold composed of cdc11p and sid4p. Cdc11p links the other SIN proteins to sid4p and the SPB. Homologues of cdc11p have been identified in Saccharomyes cerevisiae (Nud1p) and human cells (Centriolin). We have defined functional domains of cdc11p by analysis of deletion mutants. We demonstrate that the C-terminal end of cdc11p is necessary for SPB localisation. We also show that the N-terminal domain is necessary and sufficient for signal transduction, since tethering of this domain to the SPB will substitute for cdc11p in SIN function. PMID- 15135076 TI - Structure of human PRL-3, the phosphatase associated with cancer metastasis. AB - PRL-3, a novel class protein of prenylated tyrosine phosphatase, is important in cancer metastasis. Due to its high levels of expression in metastatic tumors, PRL 3 may constitute a useful marker for metastasis and might be a new therapeutic target. Here, we present the solution structure of the phosphatase domain of a human PRL-3 (residues 1-162) in phosphate-free state. The nuclear magnetic resonance (NMR) structure of PRL-3 is similar to that of other known phosphatases with minor differences in the secondary structure. But the conformation and flexibility of the loops comprising the active site differ significantly. When phosphate ions or sodium orthovanadate, which is a known inhibitor, are added to the apo PRL-3, the NMR signals from the residues in the active site appeared and could be assigned, indicating that the conformation of the residues has been stabilized. PMID- 15135077 TI - Rhamnogalacturonan lyase reveals a unique three-domain modular structure for polysaccharide lyase family 4. AB - Rhamnogalacturonan lyase (RG-lyase) specifically recognizes and cleaves alpha-1,4 glycosidic bonds between L-rhamnose and D-galacturonic acids in the backbone of rhamnogalacturonan-I, a major component of the plant cell wall polysaccharide, pectin. The three-dimensional structure of RG-lyase from Aspergillus aculeatus has been determined to 1.5 A resolution representing the first known structure from polysaccharide lyase family 4 and of an enzyme with this catalytic specificity. The 508-amino acid polypeptide displays a unique arrangement of three distinct modular domains. Each domain shows structural homology to non catalytic domains from other carbohydrate active enzymes. PMID- 15135078 TI - Characterization and promoter analysis of the mouse nestin gene. AB - The intermediate filament protein nestin is expressed in the neural stem cells of the developing central nervous system (CNS). Promoter analysis revealed that the minimal promoter of the mouse nestin gene resides in the region -11 to +183 of the 5'-non-coding and upstream flanking region, and that two adjacent Sp1-binding sites are necessary for promoter activity. Electrophoretic mobility-shift assays (EMSA) and supershift assays showed that Sp1 and Sp3 proteins selectively bind to the upstream Sp1 site. These results demonstrate an important functionality of Sp1 and Sp3 in regulating the expression of the mouse nestin gene. PMID- 15135079 TI - Stimulation of Fe-S cluster insertion into apoFNR by Escherichia coli glutaredoxins 1, 2 and 3 in vitro. AB - The oxygen sensor fumarate nitrate reductase regulator (FNR) of Escherichia coli contains in the active (anaerobic) state a [4Fe-4S](2+) cluster which is lost after exposure to O(2). In aerobically prepared apoFNR, or in FNR obtained by treatment of [4Fe-4S] . FNR with O(2) in vitro, intramolecular cysteine disulfides are found, including the cysteine residues which serve as ligands for the Fe-S cluster. It is shown here that the reconstitution of [4Fe-4S] . FNR from this form of aerobic apoFNR was preceded by a long lag phase when glutathione was used as the reducing agent. Addition of E. coli glutaredoxins (Grx) 1, 2 or 3 decreased the lag phase greatly and stimulated the reconstitution rate slightly (about twofold). Reconstitution of anaerobically prepared apoFNR, which has a lower cysteine disulfide content, showed only a short lag phase, which further decreased in the presence of Grx. It is concluded that in the lag phase the cysteine disulfides of apoFNR become reduced for the incorporation of the [4Fe 4S] cluster and that this reaction is stimulated by Grx. Thioredoxin (Trx) 1 showed no stimulation of FNR reconstitution in vitro. It is suggested that the function of Grx might be of significance for the insertion of FeS cluster in proteins containing disulfides. PMID- 15135080 TI - Steroid hormone biotransformation and xenobiotic induction of hepatic steroid metabolizing enzymes. AB - Normal reproductive development depends on the interplay of steroid hormones with their receptors at specific tissue sites. The concentrations of hormone ligands in the circulation and at target sites are maintained through coordinated regulation on steroid biosynthesis and degradation. Changed bioavailability of steroids, through alteration of steroidogenesis or biotransformation rates, leads to changes in endocrine function. Steroid hormones lose their receptor reactivity in most cases when they are bound to binding proteins, while metabolic conversion can result in either active or inactive metabolites. Hydroxylation by cytochrome P450 (CYP) enzymes and conjugation with glucuronide and sulfate are among the major hepatic pathways of steroid inactivation. The expression of these biotransformation enzymes can be induced by many xenobiotics. The barbiturate phenobarbital and the environmental toxicant 1,1-dichloro-2,2-bis(p chlorophenyl)ethylene (DDE) are among the well characterized inducers for the CYP 2B and 3A enzymes and selected conjugation enzymes. The induction of the steroid biotransformation enzymes is partly mediated through the activation of a group of nuclear receptors including the glucocorticoid receptor, the constitutive androstane receptor (CAR), the pregnane X receptor (PXR), and the peroxisome proliferator activated receptors (PPAR). Drug or chemical-induced increases in hepatic enzyme activities are often a basis for drug-drug interactions that lead to enhanced elimination and reduced therapeutic efficacy of steroidal drugs. The effects of enzyme induction on endogenous steroid clearance, along with its possible consequence, are less well understood. While enzyme induction by xenobiotics may increase clearance of the endogenous steroid, regulatory mechanisms for steroid homeostasis may adapt and compensate for altered clearance. PMID- 15135081 TI - Regulation of cytochrome P450 gene expression in the olfactory mucosa. AB - The mammalian olfactory mucosa (OM) is unique among extrahepatic tissues in having high levels, and tissue-selective forms, of cytochrome P450 (CYP) enzymes. These enzymes may have important toxicological implications, as well as biological functions, in this chemosensory organ. In addition to a tissue selective, abundant expression of CYP1A2, CYP2A, and CYP2G1, some of the OM CYPs are also known to have an early developmental expression, a resistance to xenobiotic inducers, and a lack of responsiveness to circadian rhythm. Efforts to fully characterize the regulation of CYP expression in the OM, and to identify the underlying mechanisms, are important for our understanding of the physiological functions and toxicological significance of these biotransformation enzymes, and may also shed unique light on the general mechanisms of CYP regulation. The aim of this mini-review is to provide a summary of current knowledge of the various modes of regulation of CYPs expressed in the OM, an update on our mechanistic studies on tissue-selective CYP expression, and a review of the literature on xenobiotic inducibility of OM CYPs. Our goal is to stimulate further studies in this exciting research area, which is of considerable importance, in view of the constant exposure of the human nasal tissues to inhaled, as well as systemically derived, chemicals, the prevalence of olfactory system damage in individuals with neurodegenerative diseases, and the current uncertainty in risk assessments for potential olfactory toxicants. PMID- 15135082 TI - Therapeutic efficacy of lipoic acid in combination with dimercaptosuccinic acid against lead-induced renal tubular defects and on isolated brush-border enzyme activities. AB - The combined therapeutic potentials of lipoic acid and dimercaptosuccinic acid were compared against their sole administrations in restoring the altered lead sensitive indices in urine and isolated renal brush-border preparations. Toxicity was induced in male albino rats (Wistar strain) by administering lead acetate (0.2%) in drinking water for 5 weeks, followed by therapy comprising lipoic acid (25 mg/kg body weight) and dimercaptosuccinic acid (20 mg/kg body weight) solely as well as combined during the 6th week. Changes in kidney weights encountered upon lead administration improved after therapy with lipoic acid and dimercaptosuccinic acid. Renal integrity was assessed by measuring the activities of alkaline phosphatase, acid phosphatase, lactate dehydrogenase, leucine aminopeptidase, N-acetyl-beta-D-glucosaminidase, gamma-glutamyl transferase and beta-glucuronidase in urine along with some urinary constituents (urea, uric acid, creatinine, protein and phosphorous). The effects of lead were also studied on isolated brush-border enzymes (alkaline phosphatase, acid phosphatase, gamma glutamyl transferase and beta-glucuronidase) that showed a decline upon its administration. Increased activities of urinary enzymes were accompanied by increase in the urinary constituents. Increase in renal lead content was paralleled by a drastic fall in the renal delta-aminolevulinic acid dehydratase and a rise in urinary lead levels. Relative to the administration of lead, the combined therapy showed betterment on the renal integrity with respect to the functional parameters assessed, thereby indicating its efficacy over the monotherapies. PMID- 15135083 TI - Bisphenol A and its methylated congeners inhibit growth and interfere with microtubules in human fibroblasts in vitro. AB - Bisphenol A (BPA), a monomer of polycarbonate plastics and epoxy resins, has previously been reported to induce micronuclei containing whole chromosomes in Chinese hamster V79 cells. In the present study, the aneuploidogenic potential of BPA was investigated in cultured human AG01522C fibroblasts. In contrast to the known aneugens diethylstilbestrol (DES) and 17beta-estradiol, which caused mitotic arrest and the induction of kinetochore-positive micronuclei, BPA did not induce micronuclei and inhibited the proliferation of AG01522C cells in G2 phase and probably also in G1 phase. Fluorescence microscopy of the BPA-treated cells after immunofluorescent staining of microtubules revealed structural abnormalities of the cytoplasmic microtubule complex (CMTC): densely stained rings and loops of tubulin were observed, which increased in number with increasing BPA concentration and were more stable against low temperature than normal microtubules. The mechanisms of the growth inhibition and the interference with microtubules elicited by BPA in AG01522C cells are presently unknown. The formation of rings and loops in the CMTC of AG01522C cells was also observed with two congeners of BPA carrying one and two, respectively, additional methyl groups in ortho-position to the phenolic hydroxyl group at each aromatic ring. However, in contrast to BPA itself, these congeners of BPA behaved "DES-like" by inducing mitotic arrest and kinetochore-positive micronuclei in AG01522C cells. PMID- 15135084 TI - Penta-acetyl geniposide-induced C6 glioma cell apoptosis was associated with the activation of protein kinase C-delta. AB - Herbal medicine has been utilized to treat a variety of diseases, including cancer. On the other hand, disturbance of apoptosis is often observed in cancer cells. It has been reported that protein kinase C (PKC) isoforms are involved in the signaling of apoptosis. In the present study, we investigate the antitumor effect and possible mechanism of a herbal-originated product, (Ac)(5)GP. We demonstrate that (Ac)(5)GP treatment results in DNA fragmentation of C6 glioma cells dose-dependently. Stimulated by (Ac)(5)GP, PKCdelta and PKCzeta were activated and translocated to the cell membrane fraction. Flow cytometry analysis showed that PKCdelta, but not PKCzeta inhibition blocks the (Ac)(5)GP-induced apoptosis by decreasing the cell population of sub G1 peak. However, the mRNA levels of PKCdelta and PKCzeta were not altered by (Ac)(5)GP-induced glioma cell apoptosis. These results suggested that the treatment of (Ac)(5)GP induces apoptosis of tumor cells through the activation but not the synthesis of PKCdelta. PMID- 15135085 TI - Quantitative structure toxicity relationships for catechols in isolated rat hepatocytes. AB - One- and two-parameter quantitative structure toxicity relationship (QSTR) equations were obtained to describe the cytotoxicity of isolated rat hepatocytes induced by 23 catechols in which LD(50) represents the catechol concentration required to induce 50% cytotoxicity in 2 h. A QSTR equation logLD(50) (microM = - 0.464(+/-0.065) log P + 3.724(+/-0.114) (n = 20, r(2) = 0.740, s(y,x) = 0.372, P < 1 x 10(-6), outliers: 4-methoxycatechol, 3-methoxycatechol, L-dopa) was derived where logP represents octanol/water partitioning. Outliers were determined by adopting a statistical method to standardize the identification of outliers. When pK(a1), the first ionization constant, was considered as a contributing parameter a two-parameter QSTR equation was derived: logLD(50) (microM = - 0.343(+/-0.058) log P - 0.116(+/-0.041) pK(a1)+4.389 (+/-0.315) (n = 22, r(2) = 0.738, s(y,x) = 0.375, P < 0.01, outlier: 4-methoxycatechol). Replacing logP with logD(7.4), the partition coefficient at pH 7.4, improved the first correlation by limiting the outlier to 4-methoxycatechol: logLD(50) (microM)=-0.252(+/-0.039) logD(7.4)+3.168(+/-0.090) (n = 22, r(2) = 0.671, s(y,x) = 0.420, P < 1 x 10(-5). In this study, 4-methoxycatechol (readily autooxidizable) was found to be an outlier for all QSTR equations derived. These findings point to lipophilicity and pK(a1) as two important characteristics of catechols that can be used to predict their cytotoxicity towards isolated rat hepatocytes. The catechols with the higher lipophilicity/distribution coefficient, the lower degree of ionization and the higher pK(a(catechol)) were more toxic towards hepatocytes than the other catechols. PMID- 15135086 TI - Cisplatin-evoked DNA fragmentation in normal and cancer cells and its modulation by free radical scavengers and the tyrosine kinase inhibitor STI571. AB - Cis-diamminedichloroplatinum(II) (cisplatin, cis-DDP) is well studied anticancer drug, whose activity can be attributed to its ability to form adducts with DNA, but this drug can also form DNA-damaging free radicals, however this mechanism of cisplatin action is far less explored. Using the comet assay we studied cisplatin induced DNA damage in the presence of spin traps: DMPO and PBN, Vitamins A, C and E as well as the tyrosine kinases inhibitor STI571 in normal human lymphocytes and leukemic K562 cells. The latter cells express the BCR/ABL fusion protein, which can be a target of the tyrosine kinase inhibitor STI571. A 20 h incubation with cisplatin at 1-10 microM induced DNA cross-links and DNA fragmentation in normal and cancer cells. Cisplatin could induce intra- and interstrand DNA-DNA cross-links as well as DNA-protein cross-links. DNA damage in K562 cells was more pronounced than in normal lymphocytes. In the presence of spin traps and vitamins we noticed a decrease in the DNA fragmentation in both cell types. Co-treatment of the lymphocytes with cisplatin at 10 microM and STI571 at 0.25 microg/ml caused an increase of DNA fragmentation in comparison with DNA fragmentation induced by cisplatin alone. In the case of K562 cells, an increase of DNA fragmentation was observed after treatment with cisplatin at 1 microM. Our results indicate that the free radicals scavengers could decrease DNA fragmentation induced by cisplatin in the normal and cancer cells, but probably they have no effect on DNA cross-linking induced by the drug. The results obtained with the BCR/ABL inhibitor suggest that K562 cells could be more sensitive towards co-treatment of cisplatin and STI571. Our results suggest also that aside from the BCR/ABL other factors such as p53 level, signal transduction pathways and DNA repair processes can be responsible for the increased sensitivity of K562 cells to cisplatin compared with normal lymphocytes. PMID- 15135087 TI - In vitro metabolism of fipronil by human and rat cytochrome P450 and its interactions with testosterone and diazepam. AB - Fipronil (5-amino-1-[2,6-dichloro-4-(trifluoromethyl)phenyl]-4 [(trifluoromethyl)sulfinyl]-1H-pyrazole-3-carbonitrile) is a highly active, broad spectrum insecticide from the phenyl pyrazole family, which targets the gamma amino butyric acid (GABA) receptor. Although fipronil is presently widely used as an insecticide and acaricide, little information is available with respect to its metabolic fate and disposition in mammals. This study was designed to investigate the in vitro human metabolism of fipronil and to examine possible metabolic interactions that fipronil may have with other substrates. Fipronil was incubated with human liver microsomes (HLM) and several recombinant cytochrome P450 (CYP) isoforms obtained from BD Biosciences. HPLC was used for metabolite identification and quantification. Fipronil sulfone was the predominant metabolite via CYP oxidation. The K(m) and V(max) values for human liver microsomes are 27.2 microM and 0.11 nmol/mg proteinmin, respectively; for rat liver microsomes (RLM) the K(m) and V(max) are 19.9 microM and 0.39 nmol/mg proteinmin, respectively. CYP3A4 is the major isoform responsible for fipronil oxidation in humans while CYP2C19 is considerably less active. Other human CYP isoforms have minimal or no activity toward fipronil. Co-expression of cytochrome b(5) (b(5)) is essential for CYP3A4 to manifest high activity toward fipronil. Ketoconazole, a specific inhibitor of CYP3A4, inhibits 78% of the HLM activity toward fipronil at a concentration of 2 microM. Oxidative activity toward fipronil in 19 single-donor HLMs correlated well with their ability to oxidize testosterone. The interactions of fipronil and other CYP3A4 substrates, such as testosterone and diazepam, were also investigated. Fipronil metabolism was activated by testosterone in HLM but not in CYP3A4 Supersomes. Testosterone 6beta hydroxylation in HLM was inhibited by fipronil. Fipronil inhibited diazepam demethylation but had little effect on diazepam hydroxylation. The results suggest that fipronil has the potential to interact with a wide range of xenobiotics or endogenous chemicals that are CYP3A4 substrates and that fipronil may be a useful substrate for the characterization of CYP3A4 in HLM. PMID- 15135088 TI - Inhibition of cytochrome P450 enzymes participating in p-nitrophenol hydroxylation by drugs known as CYP2E1 inhibitors. AB - p-Nitrophenol hydroxylation is widely used as a probe for microsomal CYP2E1. Several drugs are known as CYP2E1 inhibitors because of their capability to inhibit p-nitrophenol hydroxylation. Our results suggest further participation of CYP2A6 and CYP2C19 enzymes in p-nitrophenol hydroxylation. Moreover, CYP2A6 and CYP2C19 may be considered as the primary catalysts, whereas CYP2E1 can also contribute to the hydroxylation of p-nitrophenol. Further aim of our study was to evaluate the selectivity of p-nitrophenol hydroxylase inhibitors towards cytochrome P450 enzymes. The effects of antifungals: bifonazole, econazole, clotrimazole, ketoconazole, miconazole; CNS-active drugs: chlorpromazine, desipramine, fluphenazine, thioridazine; and the non-steroidal anti-inflammatory drug: diclofenac were investigated on the enzyme activities selective for CYP2A6, CYP2C9, CYP2C19, CYP2E1 and CYP3A4. None of the drugs could be considered as a potent inhibitor of CYP2E1. Strong inhibition was observed for CYP3A4 by antifungals with IC(50) values in submicromolar range. However, ketoconazole was the only imidazole derivative that could be considered as a selective inhibitor of CYP3A4. The CNS-active drugs investigated were found to be weak inhibitors of CYP2A6, CYP2C9, CYP2C19, CYP2E1 and CYP3A4. Diclofenac efficiently inhibited CYP2C9 and to a less extent CYP3A4 enzyme. PMID- 15135089 TI - Determination of 1-hydroxypyrene in children urine using column-switching liquid chromatography and fluorescence detection. AB - This study developed an acid hydrolysis method instead of using enzyme extraction, equipped with column-switching system for the pretreatment of samples, in the determination of 1-hydroxypyrene in the urine from children and pyrene in airborne particulates. We collected both types of samples from areas near a petrochemical industry and rural areas as reference. Samples were first treated with acid hydrolysis and followed by solvent extraction prior to being injected into the separation system for the determination with high performance liquid chromatography and fluorescence. A column-switching system was on-line with a C18 separation column to remove matrix interference and obtain a stable baseline of the chromatogram. The eluent used to separate the 1-hydroxypyrene was 60% (v/v) aqueous acetonitrile solution. A fluorescence detector was used to monitor 1-hydroxypyrene at lambdaex = 348 nm and lambdaem = 388 nm, and pyrene at lambdaex = 331 nm and lambdaem = 390 nm. Both calibration graphs were linear with very good correlation coefficients (r > 0.999) and the detection limits were ca. 2pg (5ng/l). Results showed that there was a significant association between 1 hydroxypyrene levels in urine specimens and pyrene levels in airborne particulate samples (r = 0.68, P < 0.05). The average levels of pyrene in the particulates (0.18 versus 0.09ng/m3) and of 1-hydroxypyrene in urine specimens (155.9 versus 110.2ng/g creatinine) were higher for the petrochemical area than for the rural area. This method is stable and sensitive for measuring polycyclic aromatic hydrocarbons in environmental samples. PMID- 15135090 TI - Analysis of PEG 400 and 4000 in urine for gut permeability assessment using solid phase extraction and gel permeation chromatography with refractometric detection. AB - We developed a treatment of urine samples allowing the analysis of two intestinal permeability markers: polyethylene glycol (PEG) 400 (highly diffusible; basal permeability indicator) and PEG 4000 (poorly diffusible; indicator of an abnormal increase of permeability) by a unique gel permeation chromatography (GPC) with refractometric detection. Urinary PEG were extracted using a mixed-bed resin composed of C2 and C18 layers. Permeability mean values determined in 11 human healthy subjects were 24.20 +/- 9.30% and 0.12 +/- 0.08% for, respectively, PEG 400 and 4000. The percentage of the PEG 4000 permeability value to the one of PEG 400 corresponded to an intestinal permeability index (IPI) of 0.52 +/- 0.35 expressing a low diffusion of this poorly permeability marker. PMID- 15135091 TI - Simultaneous quantification of beclomethasone dipropionate and its metabolite, beclomethasone 17-monopropionate in rat and human plasma and different rat tissues by liquid chromatography-positive electrospray ionization tandem mass spectrometry. AB - A sensitive, rapid and selective liquid chromatography-positive electrospray ionization tandem mass spectrometry (LC-(ESI+)-MS-MS) method has been developed and validated for the simultaneous quantification of beclomethasone dipropionate (BDP) and its active metabolite, beclomethasone 17-monopropionate (17-BMP) in rat plasma and different tissues using fluticasone propionate (FP) as the internal standard. The method was validated over a linear range from 0.05 to 5 ng/ml for both analytes. A solid-phase extraction procedure was used for plasma samples and a liquid-liquid extraction procedure for tissues samples (lung, liver and kidney). The between-day and within-day coefficients of variation for all compounds were 73%. The intra- and inter-day variations were <15%. The validated method was applied to quantify the enantiomers of esmolol and its metabolite in human plasma for hydrolysis studies. PMID- 15135098 TI - Separation and determination of dexamethasone sodium phosphate in cochlear perilymph fluid by liquid chromatography with ultraviolet monitoring and electrospray ionization mass spectrometry characterization. AB - The method for separation and determination of dexamethasone sodium phosphate (DexP) in cochlear perilymph fluid (CPF) of cavy was developed using HPLC with ultraviolet (UV) monitoring and electrospray ionization/mass spectrometry (ESI/MS) identification. The quantitative determination of DexP in CPF was achieved by HPLC with UV detection at 245 nm. The separation was carried out on a Phenomenex ODS(3) column ( 250 mm x 4.6 mm i.d., 5 microm) with the mobile phase of acetonitrile-5mmol/l ammonium acetate (23:77 (v/v)) at a flow rate of 1.0 ml/min. DexP was baseline separated from the matrices of CPF blanks within 15 min. The linearity ranged from 0.5 to 50 microg/ml. The limit of detection was 0.10 microg/ml. The recovery ranged from 98.5 to 100.8%. The relative standard deviations (R.S.D.s) of intra- and inter-day peak area were between 0.7-1.3 and 1.2-3.5%, respectively. Both full scan MS and MS2 of DexP with positive and negative polarity were obtained and elucidated. The specific ions were chosen to characterize DexP in the CPF sample. Using the proposed HPLC-UV-ESI/MS method, the concentration of DexP in CPF samples after both vein and middle ear injections were determined, and the relationships between concentration and time were obtained. This method offered reference data for clinical investigation of DexP to cure ear diseases. PMID- 15135099 TI - Determination of A 3,4-diaminopyridine in plasma by liquid chromatography with electrochemical detection using solid-phase extraction. AB - In order to quantify a small amount of a drug, 3,4-diaminopyridine (3,4-DAP), in animal plasma samples, an analytical method was developed. It involved an extraction of 3,4-DAP and phenylephrine, used as internal standard (IS), from plasma with solid-phase extraction (SPE) on C18 cartridges. This analytical method is a hyphenated technique based on high-performance liquid chromatography with electrochemical detection (HPLC-EC) whose purpose is to obtain first a sensitive method and second a satisfying separation between 3,4-DAP and phenylephrine. The analytical method is accurate, specific, and linear between 10 and 500 g of 3,4-DAP per litre. The recovery of 3,4-DAP is estimated at 70.8% with a 95% confidence interval of (66.0 -75.6%). Intermediate precision was evaluated on three quality control samples; the intra-day precision was estimated at 13.5, 9.1, 7.8% and the inter-day precision at 17.9, 8.4, 9.3%. The limit of quantification of the method was evaluated at 10 g l-1. First toxicokinetic parameters determined on dogs plasma samples after one 3,4-DAP oral administration of 1 mg kg-1 were: Cmax=395.7 microg l-1; Tmax =15 min; t1/2=113.6 min; Clearance/F=16.8 ml kg-1 min-1 and Vd/F=2.7 l kg -1. PMID- 15135100 TI - Reversed-phase liquid chromatographic method with fluorescence detection for the simultaneous determination of albendazole sulphoxide, albendazole sulphone and albendazole 2-aminosulphone in sheep plasma. AB - A rapid and sensitive HPLC method for the simultaneous quantification of albendazole sulphoxide (ABZ-SO), albendazole sulphone (ABZ-SO2) and albendazole 2 aminosulphone (ABZ-SO2NH2) in sheep blood plasma has been developed. Plasma samples were extracted with ethyl acetate under alkaline conditions. Separation was achieved on a C18 reversed-phase analytical column, in the presence of positively- (tetra-n-butylammonium hydrogen sulphate) and negatively-charged (octanesulphonate sodium) pairing ions, while detection was performed fluorometrically. Excitation and emission wavelengths were 290 and 320 nm, respectively. Limits of quantification were defined at 39 ng/ml for ABZ-SO, 4.95 ng/ml for ABZ-SO2 and 4 ng/ml for ABZ-SO2NH2. Accuracy data, in terms of recovery efficiency showed overall values (+/- S.E.M.) of 85.6 +/- 1.0% for ABZ-SO, 100.0 +/- 1.0% for ABZ-SO2 and 89.1 +/- 0.6% for ABZ-SO2NH2. The method was successfully applied to quantitatively determine the three albendazole metabolites in plasma samples collected from sheep that had been orally administered albendazole. PMID- 15135101 TI - Liquid chromatography-tandem mass spectrometry method for the determination of tranexamic acid in human plasma. AB - A new method for the determination of tranexamic acid (TA) in human plasma using high performance liquid chromatography with tandem mass spectrometric detection was described. TA and the internal standard, methyldopa, was extracted from a 200 l plasma sample by a one-step deproteination using perchloric acid. Chromatographic separation was performed on an Xtrra MS C18 Column (2.1 mm x 100 mm, 3.5 microm) with the mobile phase consisting of 10% acetonitrile in 2 mM ammonium acetate buffer (pH 3.5) at a flow rate of 0.15 ml/min. The total run time was 5 min for each sample. Detection and quantitation was performed by the mass spectrometer using the multiple reaction monitoring of the precursor-product ion pair m/z 158 --> 95 for TA and m/z 212 --> 166 for methyldopa, respectively. The method was linear over the concentration range of 0.02-10.00 g/ml with lower limit of quantification of 0.02 microg/ml for TA. The intra- and inter-day precision was less than 11% and accuracy ranged -10.88 to 11.35% at the TA concentrations tested. The present method provides a relatively simple and sensitive assay with short turn-around time. The method has been successfully applied to a clinical pharmacokinetic study of TA in 12 healthy subjects. PMID- 15135102 TI - Simultaneous determination of catecholamines and polyamines in PC-12 cell extracts by micellar electrokinetic capillary chromatography with ultraviolet absorbance detection. AB - A method for simultaneous determination of polyamines and catecholamines in cell extracts by micellar electrokinetic capillary chromatography with UV detection at 254 nm was established at the first time. The polyamines (putrescine, spermidine and spermine) and catecholamines (dopamine, serotonin, norepinephrine and epinephrine) were extracted from PC-12 cells and were derivatized with 6 aminoquinolyl-N-hydroxysuccinimidyl carbamate. Different derivatization conditions such as temperature, ratio of derivatization reagents and incubation time were investigated to find the best reaction condition which gave the highest detection sensitivity for polyamines and catecholamines. The influence of running buffer and additives on the separation such as pH, sodium dodecyl sulfate (SDS) concentrations and various additives was also investigated. Separation was achieved within 20 min with good repeatability in a 100mM boric acid buffer containing 10mM SDS and 10mM 18-crown-6 at a pH of 9.5. The detection limit ranged from 1.0 x 10(-7) to 9.0 x 10(-7) M, which is sufficient for determination of polyamines and catecholamines in many cell extracts. This technique can be easily applied to polyamine-related anticancer drug studies or clinical follow ups after each dosage of these anticancer drugs, since these drugs not only have great inhibition on polyamine levels in blood, but also have a large influence on catecholamine levels in blood. PMID- 15135103 TI - Development of a validated liquid chromatography method for the simultaneous determination of eight fat-soluble vitamins in biological fluids after solid phase extraction. AB - In the present study, a simple and rapid reversed-phase HPLC procedure has been developed for the simultaneous determination of eight fat-soluble vitamins (retinol, menadione, menaquinone, delta-tocopherol, cholecalciferol, alpha tocopherol, alpha-tocopherol acetate and phylloquinone) in biological fluids: blood serum and urine. The analytical column, Phenomenex Luna C18 (150 mm x 4.6 mm) 3 microm, was operating at ambient temperature. Mobile phase consisted of a mixture of CH3OH-CH3CN delivered using a linear gradient, starting with a composition of 50-50% v/v and ending at 30-70% at a flow rate of 1.3 ml/min. Xanthophyll was used as internal standard (2 ng/microl). Detection and identification was performed using a photodiode array detector. Eluent monitoring was achieved at 280 nm for vitamins and 450 nm for the internal standard. However, quantitation was performed at maximum wavelength for each vitamin. Detection limits were found in the range of 1.4-6.6 ng per 20-microl injected samples, while linearity held up to 25 ng/microl. The statistical evaluation of the method was examined performing intra-day (n = 6) and inter-day calibration (n = 7) and was found to be satisfactory, with high accuracy and precision results. The biological fluids were treated using solid-phase extraction cartridges, to remove all endogenous interferences from sample matrix. The solid-phase extraction protocol was optimized in terms of retention and elution. High extraction recoveries from biological matrices: blood serum and urine, (average recovery ranging between 95 and 97.6% for blood serum and between 94.2 and 95.8% for urine) were achieved for the eight fat-soluble vitamins, using Cyclohexyl J.T. Baker SPE cartridges with methanol as eluent, requiring small volumes, 100 microl of blood serum and 100 microl of urine. PMID- 15135104 TI - Analysis of coenzyme Q(10) in human plasma by column-switching liquid chromatography. AB - A new method of determining coenzyme Q10 in human plasma was developed based on column-switching high performance liquid chromatography (HPLC). CoQ10 was quantitatively extracted into 1-propanol with a fast one-step extraction procedure, after centrifugation, the supernatant was cleaned on an octadecyl bonded silica column and then transferred to reversed-phase column by a column switching valve. Determination of CoQ10 was performed on a reversed-phase analytical column with ultraviolet detection at 275 nm and the mobile phase containing 10% (v/v) isopropanol in methanol at a flow-rate of 1.5 ml/min. The sensitivity of this method allows the detection of 0.1 microg/ml CoQ10 in plasma (S/N=3). The linearity between the concentration and peak height is from 0.05 to 20 mg/l. The reproducibility (R.S.D.%) of the method is less than 2% (within day) and less than 3% (between day), the average recovery is 100.9 + 2.1%, it takes only 30 min to complete an analysis procedure, suitable for the determination of CoQ10 in human plasma especially for batch analysis in clinical laboratories. Finally, the method was applied to determine the plasma CoQ10 levels in healthy subjects, hyperthyroid and hypothyroid patients. PMID- 15135105 TI - Determination of chloralose residues in animal tissues by liquid chromatography electrospray ionisation tandem mass spectrometry. AB - A relatively rapid and specific method for the determination of chloralose in animal tissues by LCMSMS was developed. Isocratic reverse phase HPLC was used to introduce samples for electrospray negative ionisation tandem mass spectrometry. Methanol extracts were diluted to approximate the mobile phase composition, then filtered prior to analysis. Residues were identified by monitoring the multiple reaction monitoring (MRM) transitions of precursor ions mass:charge (m/z) 309 and 307 to a common m/z 161 product ion. Qualitative and quantitative confirmation data were acquired simultaneously by monitoring alternative MRM transitions. Calibration was linear over a working range of 0.025-1.3 microg/ml, and the limit of quantitation (LOQ) was 0.28 mg/kg for liver. The mean recovery was 88.5% from chicken muscle tissue fortified at 198-237 mg/kg, and ranged from 81.3 to 94.3% from liver tissue fortified at 1-52 mg/kg. The method is compared to a gas chromatography (GC) procedure previously employed. PMID- 15135106 TI - Determination of benidipine in human plasma using liquid chromatography-tandem mass spectrometry. AB - We developed a method for determining benidipine, a dihydropyridine analogue calcium-channel blocker, in plasma using liquid chromatography-tandem mass spectrometry (LC-MS-MS). Benidipine and benidipine-d5, an internal standard, were extracted from plasma using diethyl ether in the presence of 5M NaOH. After drying the organic layer, the residue was reconstituted in acetonitrile and injected onto a reversed-phase C18 column. The isocratic mobile phase (acetonitrile-5mM ammonium acetate, 90:10, v/v) was eluted at 0.2 ml/min. The ion transitions monitored in multiple reaction-monitoring mode were m/z 506-174 for benidipine and m/z 511-179 for the internal standard. The coefficient of variation of the assay precision was less than 13%, and the accuracy exceeded 92%, except at the limit of quantification, 0.05 ng/ml with 1ml of plasma, when it was 85%. This method was used to measure the benidipine concentration in plasma from healthy subjects after a single 4-mg oral dose of benidipine. This method is a very simple, sensitive, and accurate way to determine the plasma benidipine concentration. PMID- 15135107 TI - Affinity membrane chromatography: relationship of dye-ligand type to surface polarity and their effect on lysozyme separation and purification. AB - Two different dye-ligands, i.e. Procion Brown MX-5BR (RB-10) and Procion Green H 4G (RG-5) were immobilised onto poly(2-hydroxyethylmethacrylate) (pHEMA) membranes. The polarities of the affinity membranes were determined by contact angle measurements. Separation and purification of lysozyme from solution and egg white were investigated. The adsorption data was analysed using two adsorption kinetic models the first order and the second order to determine the best-fit equation for the separation of lysozyme using affinity membranes. The second order equation for the adsorption of lysozyme on the RB-10 and RG-5 immobilised membranes systems is the most appropriate equation to predict the adsorption capacity for the affinity membranes. The reversible lysozyme adsorption on the RB 10 and RG-5 did not follow the Langmuir model, but obeyed the Temkin and Freundlich isotherm model. Separation and purification were monitored by determining the lysozyme activity using Micrococcus lysodeikticus as substrate. The purities of the eluted lysozyme, as determined by HPLC, were 76 and 92% with recovery 63 and 77% for RB-10 and RG-5 membranes, respectively. For the separation and purification of lysozyme the RG-5 immobilised membrane provided the best results. The affinity membranes are stable when subjected to sanitization with sodium hydroxide after repeated adsorption-elution cycles. PMID- 15135108 TI - Determination of l-arginine and NG, NG - and NG, NG' -dimethyl-L-arginine in plasma by liquid chromatography as AccQ-Fluor fluorescent derivatives. AB - A new HPLC assay for the detection of L-arginine, NG, NG-dimethyl-L-arginine (ADMA) and NG, NG' -dimethyl-L-arginine (SDMA) in plasma using the derivatisation reagent AccQ-Fluor (6-aminoquinolyl-N-hydroxysuccinimidyl carbamate) is described. The fluorescent derivatives produced are extremely stable enabling routine processing of large numbers of samples. Arginine and its metabolites are extracted from plasma on strong cation exchange (SCX) cartridges with NG monomethyl-L-arginine (NMMA) as internal standard, derivatised and separated on a C18 column with acetonitrile in 0.1M sodium acetate buffer pH 6. Separation of the stereoisomers ADMA and SDMA was excellent and improvements to the solid phase extraction (SPE) procedure enabled good recovery (>80%) of arginine, ADMA and SDMA. The utility of the method is exemplified by comparison of plasma concentrations of ADMA, SDMA and arginine in healthy volunteers and diabetic/ischaemic patients. PMID- 15135109 TI - Optimisation of the separation of four major neutral glycosphingolipids: application to a rapid and simple detection of urinary globotriaosylceramide in Fabry disease. AB - A simple method for the separation of the four major neutral glycosphingolipids, present in all human tissue, was developed. This gradient normal phase-HPLC method utilises a polyvinyl alcohol bonded stationary phase and an evaporative light-scattering detection (ELSD). Screening pure solvents in a binary gradient elution mode allowed, in a first step, to assess the behaviour of the studied solutes and to select the solvents for further mobile phase optimisation. The proportion of the remaining solvents was defined to reach a maximal resolution. The reduction of the analysis time and the enhancement of the signal were obtained by optimising the gradient slope and the flow-rate. Optimal levels of triethylamine and formic acid (TEA-FA) for the enhancement of the evaporative light scattering detector response were established at 0.1% (v/v). Thus, the optimal conditions for the separation of the four glycosphingolipids was obtained with a gradient elution from a 100% chloroform to a 100% acetone:methanol (90:10 (v/v)) mobile phase at 0.2 ml min-1, using a 10% min-1 gradient slope. Finally, this method was applied to detect the excess of one of the neutral sphingolipids, namely globotriaosylceramide (Gb3) in the urine of patients affected with Fabry disease. A liquid-liquid extraction of the sediments obtained from an aliquot of only ten ml of urine proved sufficient to detect the excess of Gb3 present in both hemizygote and heterozygote patients. In all, the ability of our method to detect abnormal amounts of Gb3 in urinary sediments could allow the diagnosis of weakly symptomatic Fabry patients in large screening programs PMID- 15135110 TI - Determination of the deoxycytidine kinase activity in cell homogenates with a non radiochemical assay using reversed-phase high performance liquid chromatography; Identification of a novel metabolite of 2-chlorodeoxyadenosine. AB - A non-radioactive procedure to measure the deoxycytidine kinase (dCK) activity in crude cell free homogenates was developed. 2-Chlorodeoxyadenosine (CdA) was used as the substrate for dCK and was separated from its product 2 chlorodeoxyadenosine-5'-monophosphate (CdAMP) by reversed-phase HPLC. A complete separation of CdA and its metabolites was achieved in 30 min. The minimum amount of CdAMP that could be detected was 1 pmol. The assay was linear with reaction times up to at least 3h. With respect to the protein concentration, the reaction was linear with protein concentrations up to 760 microg/ml in the assay. An amount of 8 x 10(3) cells was already sufficient to determine the specific dCK activity in SK-N-BE(2)c cells. CdA was not only converted to CdAMP but also to 2 chloroadenine and, surprisingly, also to 2-chlorodeoxyinosine, in MOLT-3 cells. The deamination of CdA was completely inhibited by deoxycoformycin, which clearly demonstrates that CdA is a substrate for adenosine deaminase. PMID- 15135111 TI - High-performance liquid chromatographic assay of lactic, pyruvic and acetic acids and lactic acid stereoisomers in calf feces, rumen fluid and urine. AB - To facilitate clinical investigation of metabolic acidosis, a high-performance liquid chromatographic method was adapted and validated for the chiral separation of D-(-) and L-(+)-lactic acid in calf feces, rumen fluid and urine. A non-chiral method was also adapted and validated for the separation of pyruvic, acetic and DL-(+/-)-lactic acids in calf feces and DL-(+/-)-lactic and pyruvic acids in rumen fluid. Separation and quantification were achieved using a reversed phase sulphonated polystyrenedivinylbenzene analytical column for pyruvic, acetic and racemic lactic acids and by a 3 microm octadecylsilane (ODS) packed analytical column coated with N,N-dioctyl-L-alanine as the chiral selector for the separation of lactic acid enantiomers with Cu(II)-containing eluents by stereoselective ligand exchange chromatography. Endogenous analytes were present in validation samples over a range of concentrations (0.2-14.8 mmol/l). For the stereoselective assay, mean intra-day accuracy ranged from 90.6 to 108.4% and intra-day precision from 0.3 to 13.8%. For the non-stereoselective assay, mean intra-day accuracy ranged from 90.4 to 108.8% and intra-day precision from 1.5 to 11.1%. The limit of quantitation was 1.0 mmol/l for D- and L-lactic acid, 0.06125 mmol/l for pyruvic acid, 1.0 mmol/l for DL-lactic acid and 1 mmol/l for acetic acid. These assays can be used to study the role of the gastrointestinal tract and kidney in metabolic acidosis. PMID- 15135112 TI - Liquid chromatographic assay for dicloxacillin in plasma. AB - A simple high-performance liquid chromatographic method for the determination of dicloxacillin in plasma has been developed. The method only requires 0.5 ml of plasma, phosphate buffer solution (pH = 4.7), acidification with 0.5N hydrochloride acid and liquid extraction with dichloromethane. Posterior evaporation of organic under nitrogen steam and redissolution in mobile phase is carried out. The analysis was performed on a Spherisorb C18 (5 microm) column, using methanol -0.05 M phosphate buffer, pH = 4.7 (75:25; v/v) as mobile phase, with ultraviolet detection at 220 nm. Results showed that the assay is sensitive: 0.5 microg/ml. The response is linear in the range of 0.5 - 10 microg/ml. Maximum inter-day coefficient of variation was 12.4%. Mean extraction recovery obtained was 96.95%. Stability studies showed that the loss was not higher than 10%, samples are stable at room temperature for 6 h, at -20 Celsius for 2 months, processed samples were stable at least for 24 h and also after two freeze-thaw cycles. The method has been used to perform pharmacokinetic and bioequivalence studies in humans. PMID- 15135113 TI - Quantitative determination of astilbin in rabbit plasma by liquid chromatography. AB - A simple method for determining the concentration of astilbin, a flavanone, in rabbit plasma has been developed. After liquid-liquid extraction, the flavanone was detected by HPLC on a 4.6-microm octadecylsilica column (Nova-Pak C-18) at 291 nm. Linear calibration graphs for astilbin were constructed from 0.44 to 22.17 microM. The limit of quantitation was 0.44 microM in plasma. The method has been applied to pharmacokinetic studies after a single i.v. and an oral administration of the compound to rabbits. PMID- 15135114 TI - Affinity adsorbent based on combinatorial phage display peptides that bind alpha cobratoxin. AB - Combinatorial phage display was used to discover peptides that selectively bind to the alpha-cobratoxin (neurotoxin) component of the multi-component venom of the Thai cobra, Naja kaouthia. Peptide sequences determined in this way were synthesized chemically and were covalently attached to agarose through the alpha amino terminus. Such affinity chromatography supports selectively bound the alpha cobratoxin component from crude venom, while passage of the crude venom over the support selectively depleted the venom of this component. The selective binding of alpha-cobratoxin to peptide-based solid-phase supports suggests that a limitless variety of peptides similarly obtained by combinatorial phage display can be used to craft specific analytical and preparative tools. PMID- 15135115 TI - Determination of paraldehyde by gas chromatography in whole blood from children. AB - A rapid, sensitive and selective gas chromatographic method with flame ionization detection was developed for the determination of paraldehyde in small blood samples taken from children. Whole blood samples (300 microl) collected in a 3 ml Wheaton glass sample vial were spiked with acetone (internal standard: 15 ng) followed by addition of concentrated hydrochloric acid. The mixture was heated in the sealed airtight sample vial in a water bath (96 Celsius; 5 min) to depolymerize paraldehyde to acetaldehyde. A 2 ml aliquot of the headspace was analyzed by gas chromatography with flame ionization detector using a stainless steel column (3 m x 4 mm i.d.) packed with 10% Carbowax 20 M/ 2% KOH on 80/100 Chromosorb WAW. Calibration curves were linear from 1.0-20 microg (r2>0.99). The limit of detection was 1.5 microg/ml, while relative mean recoveries at 2 and 18 microg were 105.6 +/- 8.4 and 101.2 +/- 5.9%, respectively (n = 10 for each level). Intra- and inter-assay relative standard deviations at 2, 10 and 18 microg were <15%. There was no interference from other drugs concurrently used in children with severe malaria, such as anticonvulsants (diazepam, phenytoin, phenobarbitone), antipyretics/analgesics (paracetamol and salicylate), antibiotics (gentamicin, chloramphenicol, benzyl penicillin) and antimalarials (chloroquine, quinine, proguanil, cycloguanil, pyrimethamine and sulfadoxine). The method was successfully applied for pharmacokinetic studies of paraldehyde in children with convulsions associated with severe malaria. PMID- 15135117 TI - The pathogenesis of interstitial lung diseases in children. AB - The pathological changes observed in interstitial lung disease (ILD) are characterised by derangements of the alveolar walls. For a long time, the prevailing hypothesis has emphasised the key role of a persistent alveolitis that injures the lung and modulates fibrogenesis, regardless of initiating agents. The current concept on ILD pathogenesis relies on an epithelial/fibroblastic pathway with epithelial injury and activation, formation of subepithelial fibroblast/myofibroblast foci and excessive accumulation of extracellular matrix. An essential step in the restoration of alveolar integrity is the rapid re epithelialisation of the altered surface, mainly through epithelial proliferation and migration. In the context of lung growth and development in paediatric ILD, it is suggested that the programmed production of mitogenic factors may promote the process of re-epithelialisation and may help to counteract the altered secretion of mediators involved in migration and proliferation of fibroblasts and differentiation into myofibroblasts. This is supported by clinical observations indicating that paediatric ILD is more responsive to therapeutic strategies than adult ILD. PMID- 15135118 TI - Clinical presentation of interstitial lung disease in children. AB - Interstitial lung disease (ILD) in children represents a heterogeneous group of rare lung disorders characterised by an inflammatory process of the alveolar wall and the pulmonary interstitium, which share common clinical, radiological and histological findings. Clinical presentation of paediatric ILD is non-specific and variable. Respiratory signs and symptoms include tachypnoea, rapid shallow breathing, retractions, cyanosis and crackles. Failure to thrive and respiratory distress are common in infants. Dyspnoea on exertion is a symptom encountered in older children. Non-respiratory symptoms can be present when ILD is part of a systemic disorder. Although non-specific and variable, the clinical presentation at the time of diagnosis has some prognostic value. PMID- 15135119 TI - Diagnosis of interstitial lung disease in children. AB - Chronic interstitial lung disease (ILD) in infants and children is a challenging diagnostic clinical problem. There are many unresolved and controversial issues in the diagnosis of this heterogeneous group of uncommon disorders in children. Diagnosis requires a high index of suspicion as the initial clinical manifestations are subtle, highly variable and non-specific. There is no consensus for the clinical diagnostic criteria of paediatric ILD. The spectrum of clinical findings is highly variable. The diagnostic evaluation of a child with suspected ILD includes a comprehensive history, physical examination, oxygen saturation (at rest, during exercise or during feeding), a plain chest x ray and a high-resolution thin-cut tomography scan of the chest. Pulmonary function studies can be useful in older children; these typically show a restrictive pattern with a decreased forced expiratory volume in 1 s (FEV(1)), forced vital capacity (FVC) and total lung capacity, but normal FEV(1)/FVC. A systematic approach to diagnosis is useful in the evaluation of an infant or child with suspected chronic ILD. Due to the rarity of most of these disorders, multi-centre collaboration is needed to improve our understanding of this orphan lung disease. PMID- 15135120 TI - Treatment of interstitial lung disease in children. AB - The treatment of interstitial lung disease in children depends on the nature of the underlying pathology. In approximately 50% of cases a specific aetiology can be found such as: chronic viral infection, an auto-immune process, sarcoidosis or alveolar proteinosis. In the remainder, the process is idiopathic and the pathological findings are based on the descriptive morphological features seen in the diagnostic lung biopsy. If a specific cause is found then targeted treatment with antivirals, steroids or other immunosuppressive agents is available. Alveolar proteinosis can be treated by bronchial lavage and GM-CSF. Idiopathic cases are treated primarily with intravenous pulsed methylprednisolone or oral prednisolone backed up hydroxychloroquine. Other immunosuppressive agents such as azathioprine, methotrexate or ciclosporin have been used successfully in individual patients. The prognosis is very variable and includes no response to any therapy, partial response with chronic long term morbidity, to virtually complete recovery. The overall mortality rate is 15%. There are no controlled therapeutic trials available because of the rarity of these conditions in childhood. Unlike in adult practice, no correlation has as yet been demonstrated between the initial pattern of chest x-ray change or the degree of pathological change on the lung biopsy and the clinical outcome. The recurrence rate within families is 1 in 8. PMID- 15135121 TI - Phenotype-specific treatment of difficult asthma in children. AB - Most children with asthma can be treated successfully with low-to-moderate doses of inhaled corticosteroid and long-acting beta-2 agonist. Those that fail to respond are a heterogeneous group. We propose that the nature and type of any steroid-resistant inflammation, the extent of any persistent airflow limitation and the extent of bronchial hyper-reactivity should be determined separately to allow a rational treatment approach to these children, rather than the haphazard advice of many current guidelines. Reasons for persistent difficult asthma include persistent eosinophilic inflammation, non-eosinophilic inflammation, airway reactivity without residual inflammation and persistent airflow limitation. We propose a protocol that uses non-invasive and invasive (bronchoscopic) methods to document the response to systemic steroids (depot triamcinolone). The aim of the protocol is to determine an individualised treatment plan; for example, cyclosporin for persistent eosinophilic inflammation, azithromycin for persistent neutrophilic inflammation and continuous subcutaneous terbutaline if there is airway reactivity without residual inflammation. Multi-centre studies are required to test the utility of this approach. PMID- 15135122 TI - Transition of young people with respiratory diseases to adult health care. AB - As many more young people with chronic health conditions survive well into adulthood, transition of affected young people from paediatric to adult health care is increasingly becoming the expected standard of care. Despite this, there is still little objective or long-term evidence to guide the development or deployment of developmentally appropriate transition processes or to define the precise manner in which the transfer of care should take place. Nevertheless, certain principles of transition have now received nearly universal endorsement. In this paper, the rationale for transition is discussed, practical strategies for transition are described, the existing evidence base supporting transition is reviewed and limitations in our knowledge are outlined. Ultimately, a carefully planned transition to adult health care should improve self-reliance, enhance autonomy and independence and support young people in attaining their maximum potential and meaningful adult lives. PMID- 15135123 TI - Infections and other causes of death in HIV-infected children in Africa. AB - The human immunodeficiency virus (HIV) pandemic has dramatically reversed improvements in infant mortality and child survival in sub-Saharan Africa. However, accurate information on the specific causes of HIV-related morbidity and mortality arising from vertical transmission is infrequent and is constrained in resource-poor settings by infrastructure and local access to health care. Such knowledge is essential to improve clinical management of HIV-infected children in Africa. In this review, a global overview of the clinical aspects of HIV infection in children is given. Factors influencing HIV disease progression, morbidity and mortality are discussed from studies on a cohort of HIV-infected children that were followed at an orphanage in Nairobi between 1999 and 2001. These parameters are contrasted with available data on HIV-infected children residing in community settings in Africa. PMID- 15135124 TI - Implementing asthma education programmes in paediatric respiratory care: settings, timing, people and evaluation. AB - Randomised controlled trials have shown that asthma education programmes in paediatric care can improve the ability of children and their parents to control their asthma. This paper discusses the implementation of asthma education within pragmatic care. Key times for implementation are suggested: at the initial diagnosis, after acute asthma events such as hospitalisation, at the time of medication change and at regular review. Special issues, such as implementing programmes for teenagers, are discussed. Evaluation is recommended for processes (does the practice have an agreed protocol for education, does it have a key person with responsibility for co-ordinating education within the clinic or practice, does it have agreed outcomes for audit of education?). Evaluation is also recommended for individual outcomes (clinical improvement and increase in perceived asthma control and confidence in management). PMID- 15135125 TI - Tracheomalacia. AB - Tracheomalacia is a rare condition characterised by collapse of the trachea during respiration. The condition is seen most often in infants and young children. Mild cases can be managed expectantly; however, severe cases can be associated with life-threatening cyanotic attacks and intervention to stabilise the airway is invariably necessary. Most commonly this involves an aortopexy to suspend the anterior wall of the trachea but other options include endoluminal or extraluminal stenting, long-term positive pressure ventilation and tracheostomy. Although tracheomalacia resolves spontaneously in most infants within the first few years of life, severe tracheomalacia is associated with significant morbidity and mortality that should not be underestimated. PMID- 15135126 TI - Asthma phenotypes in childhood: lessons from an epidemiological approach. AB - Asthma is a heterogenous disease with variable signs and symptoms among patients. It also presents significant individual variability over time. Recently, some important population-based studies that followed children from birth or from early childhood into adulthood have shed new light on how we understand this syndrome. Three phenotypes have been identified in children with asthma: transient wheezing, non-atopic wheezing of the toddler and pre-school-aged child and IgE-mediated wheezing. Transient wheezing is associated with symptoms that are limited to the first 3-5 years of life, decreased lung function, maternal smoking during pregnancy and exposure to other siblings or children at daycare centres. There is no association between transient wheezing and family history of asthma or allergic sensitisation. Children wheezing with respiratory syncytial virus in the first years of life are more likely to be wheezing up to 13 years of age; this is independent of atopy (non-atopic wheezers) and is not related to atopic sensitisation. Wheezing associated with evidence of allergic sensitisation has been identified as the 'classic' asthma phenotype. Early allergic sensitisation is a major risk factor for persistent asthma. PMID- 15135127 TI - Natural hair patterns. AB - Hair loss is complex and can be classified in many ways. This article has focused on the anatomic zones and their significance to hair restoration. Hair restoration surgeons must not only be aware of these patterns of loss but also their significance for the future. PMID- 15135128 TI - Pharmacologic management of pattern hair loss. AB - Pharmacologic management of pattern hair loss is an active area of research, and clinicians should be aware of new data and treatment modalities. Under-standing the proper role of pharmacology as it relates to surgical hair restoration and nonmedical options is crucial to provide patients with the best clinical,aesthetic, and psychological benefits. PMID- 15135129 TI - Laser hair removal. AB - The evolution of new technologies has helped to improve the clinical efficacy of laser hair removal and increase understanding of hair biology and phototrichoregulatory interactions. Long-term hair removal is now a realistic goal in the majority of individuals. Although the optimal schedule for repeat treatments is not known, most laser surgeons report treatments at 4- to 8-week intervals or at the first signs of regrowing hairs. Newer radiofrequency technologies might address the issue of white and light blond hair phenotypes;however, their exact role in the laser hair removal armamentarium remains to be further elucidated by future studies. PMID- 15135130 TI - Principles and techniques used to create a natural hairline in surgical hair restoration. AB - The principles and techniques outlined in this article will help the surgeon create hairlines that have the high degree of naturalness and substance expected by today's discerning patients. Surgeons can now establish a natural hairline during the first surgery that can stand on its own, independent of further sessions. It is not enough to "just use micrografts."Proper selection and use of FUs combined with artistry and skill provide surgeons with the tools to follow nature's lead in creating natural-appearing,soft hairlines while establishing the illusion of density. The author's approach is to create an initial framework based on the principles of hairline design,then use artistic ability and experience to fine-tune this framework. PMID- 15135131 TI - Follicular cell implantation: an update on "hair follicle cloning". AB - Follicular cell implantation has the potential to overcome many of the limitations of current surgical hair restoration, especially the finite supply of donor hair. Years of animal research have demonstrated the soundness of the basic concept, but reports in humans have shown inconsistency and problems with reproducibility. Some of these problems include (1) identifying the ideal cell type or types of the follicle to isolate and culture, (2) determining how to keep the cells inductive throughout the culturing process, and(3) overcoming economic and regulatory hurdles to bringing a safe and effective treatment to the market place. The prospect of having an unlimited supply of donor hair available to treat hair loss will continue to spur tissue engineering-based research to overcome these hurdles. PMID- 15135132 TI - Follicular unit transplantation: dissecting and planting techniques. AB - FUT was designed to move hair from the donor area to the recipient area in such a way as to duplicate naturally occurring hair and to minimize trauma to the follicles in the process. With the exception of a few factors such as the "vellus blush" of the frontal hairline or the anterior temples, FUT can accomplish these goals. This article has outlined the basic requirements for the dissection and implantation of follicular unit grafts and provided evidence provided by research data, where it exists, to support the contentions and requirements of FUT. With adherence to defined standards, surgeons can accomplish certain aesthetic goals with this technique on a consistent basis. PMID- 15135133 TI - The donor area. AB - The donor area should be treated as a vital,nonrenewable resource. Its proper, efficient use and preservation remains the cornerstone of successful transplantation by way of grafting. Evaluation of donor hair characteristics and density are essential to proper candidate selection and surgical planning. Careful donor strip removal or careful FUE are vital to avoid transection of the follicles and to maximize the surgical result. PMID- 15135134 TI - The treatment of female pattern hair loss and other applications of surgical hair restoration in women. AB - In the specialty of surgical hair restoration, men comprise more than 90% of the patients treated; however, in the last few years the number of women undergoing the procedure has increased significantly. The reasons for this growth are many and include the increase in public awareness of the efficacy of hair transplantation from such sources as the media,the Internet, advertising and word of mouth. More importantly, advances in technique have significantly improved results, increasing the confidence level in women to undergo the procedure and in hair transplant specialists to offer it. PMID- 15135135 TI - The latest developments in surgical hair restoration. AB - Surgical hair restoration has evolved rapidly over the past 10 years. Patient outcome is paramount. Current developments in hair transplantation focus on using the most appropriate harvesting techniques to use this finite resource effectively and to relocate it quickly and efficiently to achieve the optimum effective aesthetic coverage. To date,follicular unit transplantation has afforded surgeons the greatest efficiency in hair trans-plantation. These units are unlikely to become smaller, so surgeons must look at what other means are available to improve the final result. PMID- 15135136 TI - Hair transplantation to the eyebrow, eyelashes, and other parts of the body. AB - Since its earliest applications, hair transplantation has been used for treating not only the scalp in pattern baldness but also other parts of the body, commencing with eyebrow reconstruction. The earliest micrografts were applied to the eyebrow more than 30 years before their application to the scalp became the standard of care. Today hair transplantation is applied to a number of other areas. The principle behind transplanting these areas is the same-once transplanted, the hairs continue to grow because of the phenomenon of donor dominance. This article reviews the role and technique of hair transplantation to the eyebrows and eyelids, chest, beard and moustache, and pubic escutcheon. PMID- 15135137 TI - Correcting problems in hair restoration surgery: an update. AB - Hair is an important emblem of health, youth, vigor, and vitality. The state of the art in hair restoration today is to create a result that is undetectable as a surgical hair transplant. Most procedures performed using previous techniques of plug hair trans-plantation are not aesthetically acceptable by today's standards, especially in the face of progressive hair loss, which can unmask previously camouflaged corn row plugs. A technique to reduce the plugs and recycle the grafts into smaller grafts is described. The recycled hair grafts can be combined with scalp lifting, scalp reductions, and occipital harvesting of grafts to improve the results of corn row-appearing hair trans-plants and other problems of surgical hair restoration. PMID- 15135139 TI - Expression of VEGF isoforms by epiphyseal chondrocytes during low-oxygen tension is HIF-1 alpha dependent. AB - OBJECTIVE: To establish the role of hypoxia and HIF-1 alpha for VEGF expression of murine epiphyseal chondrocytes. To analyze the effect of hypoxia on VEGF isoform expression. MATERIALS AND METHODS: VEGF mRNA and VEGF isoform expression was investigated in epiphyses of murine newborns by in situ hybridization and real-time PCR. Further, epiphyseal chondrocytes were isolated from newborn mice with homozygous flanking of the HIF-1 alpha gene with lox-P sites. HIF-1 alpha was deleted by infection with adenovirus containing cre-recombinase. After chondrocytes reached confluency they were exposed to 0.5% or 20% oxygen, respectively. Total VEGF and VEGF isoform mRNA expression levels were measured by real-time PCR. Secreted VEGF protein was determined by ELISA. RESULTS: VEGF mRNA signals were detected in the hypertrophic zone and in the center of the proliferative zone of the murine epiphysis, which is considered to be hypoxic. Real-time PCR revealed that VEGF(120)is the dominant isoform in vivo. In cultured epiphyseal chondrocytes strongly increased VEGF gene expression levels were detected after exposure to hypoxia. Furthermore, secretion of VEGF protein was significantly enhanced under 0.5% oxygen. Remarkably, functional inactivation of HIF-1 alpha abolished the hypoxic increase of VEGF expression in chondrocytes completely. Furthermore, the soluble isoforms VEGF(120)and VEGF(164)are the most abundantly expressed splice variants in chondrocytes exposed to low oxygen levels. CONCLUSIONS: The data presented here clearly indicate that hypoxia is able to induce the synthesis of soluble VEGF isoforms by epiphyseal chondrocytes, most likely through stabilization of HIF-1 alpha. Thus it can be speculated that HIF-1 alpha is an essential prerequisite for hypoxic VEGF synthesis in the epiphysis, thereby contributing to the formation and invasion of blood vessels in long bone development. PMID- 15135140 TI - Serum levels of type IIA procollagen amino terminal propeptide (PIIANP) are decreased in patients with knee osteoarthritis and rheumatoid arthritis. AB - OBJECTIVE: The aim of this study was to develop a specific immunoassay for PIIANP and measure its serum concentration in healthy controls and in patients with osteoarthritis (OA) and rheumatoid arthritis (RA). In addition, we investigated circulating forms recognized by antiserum IIA in pools of serum from healthy adults, patients with OA and patients with RA. DESIGN: Using as immunogen and standard the recombinant human Glutathione S-Transferase (GST)-exon 2 fusion protein of type II collagen, we developed a competitive polyclonal antibody-based ELISA. We compare serum PIIANP levels in 43 patients with knee OA (23 women and 20 men; mean age: 62.6+/-9.6 yr), 63 women with RA (mean age: 54+/-16 yr) and 88 healthy controls (67 women, mean age: 53+/-13 yr and 21 men, mean age: 63+/-7 yr). We randomly selected serum in each group for analyze circulating forms. RESULTS: The immunoassay we developed demonstrated adequate intra and inter-assay precision (CV<10%) and dilution recovery (mean: 96%), allowing accurate measurements of serum PIIANP from 1.13 to 40 ng/ml. No significant cross reactivity of the ELISA was observed with purified intact human procollagen type I N-propeptide, circulating thrombospondin and von Willebrand factor, proteins which exhibit significant sequence homology with PIIANP. Western blot analysis showed that antiserum IIA recognized two circulating immunoreactive forms of approximately 80 and 100 KDa respectively in serum from healthy adults, patients with OA and RA but also in a pool of synovial fluids from patients with OA. Serum PIIANP levels were markedly decreased in patients with knee OA (12.0+/-3.2 vs 25.8+/-7.5 ng/ml for OA and controls respectively, P<0.0001) and RA (14.1+/-2.5 ng/ml vs 21.7+/-7.6 ng/ml for RA and controls respectively, P<0.0001). In patients with RA, serum PIIANP levels were higher in those taking low-dose prednisone compared to non-users (15.0+/-2.4 vs 13.5+/-2.4 ng/ml, P<0.05). CONCLUSIONS: We have developed the first specific immunoassay for serum PIIANP which exhibits adequate technical performances. This assay detects specifically two immunoreactive forms both in healthy adults and patients with arthritis and does not cross react with other proteins with sequence homology with PIIANP. Levels of PIIANP were significantly decreased in patients with knee OA and RA suggesting that type IIA collagen synthesis may be altered in these arthritic diseases. The measurement of type IIA collagen synthesis with this new molecular marker may be useful for the clinical investigation of patients with joint diseases. PMID- 15135141 TI - Loss of chondrogenic potential in dedifferentiated chondrocytes correlates with deficient Shc-Erk interaction and apoptosis. AB - OBJECTIVE: If dedifferentiated chondrocytes could be induced to redifferentiate in vitro, then we might thereby be furnished with a population of phenotypically stable cells for autologous implantation in reconstructive surgery. We therefore investigated the redifferentiation capabilities of chondrocytes which, having migrated from alginate beads to form a monolayer, were subsequently passaged. We also characterized the molecular traits of irreversibly dedifferentiated cells. METHODS: Human chondrocytes that had migrated from alginate beads to form a monolayer (passage 1) were passaged seven times (passages 2-8). Cells from each passage were then recultivated in alginate beads. We assessed the synthesis of type-II collagen, cartilage-specific proteoglycans, adhesion molecules (integrins), signaling proteins (Src-homology collagen [Shc] and extracellular signal-regulated kinase [Erk]) and the apoptosis marker 'activated' caspase-3 in monolayer or secondary alginate cultures. RESULTS: The synthesis of cartilage specific type-II collagen, alpha 3-integrin, Shc and activated Erk1/2 decreased rapidly after four passages in monolayer culture. Up to passage 4, cells redifferentiated in alginate culture. However, between passages 5 and 8, cells began to produce activated caspase-3; these cells not only failed to redifferentiate when recultivated in alginate, but underwent apoptosis. CONCLUSION: We conclude that the loss of chondrogenic potential by chondrocytes maintained in monolayer culture is associated with a decrease in the synthesis of cartilage markers and with a suppressed activation of key signaling proteins in the Ras-mitogen-activated protein kinase pathway (Shc and Erk1/2). These events lead to apoptosis. A decrease in Shc/Erk expression/interaction could serve as a recognition marker for irreversibly dedifferentiated chondrocytes in tissue engineering. PMID- 15135142 TI - Translation, adaptation and validation of the Western Ontario and McMaster Universities osteoarthritis index (WOMAC) for an Arab population: the Sfax modified WOMAC. AB - OBJECTIVE: To translate into Arabic and validate the Western Ontario and McMaster Universities (WOMAC) index. METHOD: Arabic translation was obtained with use of the forward and backward translation method. Adaptations were made after a pilot study. Patients with symptomatic knee OA fulfilling the revised criteria of the American College of Rheumatology were included. Impairment outcome measures (pain as measured on a visual analog scale, the maximum distance walked, Kellgren's radiological score), Lequesne index score and Beck depression scale score were recorded. Each item was analyzed. Test-retest reliability was assessed with use of the intra-class correlation coefficient (ICC) and the Bland and Altman method. Construct validity was investigated with use of Spearman's rank correlation coefficient, and a factor analysis was performed. RESULTS: One hundred and three patients were included in the study. Eight questions of the WOMAC physical function subscale (PF) had insufficient psychometric properties and were excluded. Although test-retest reliability of the questionnaire was good (0.84, 0.84, and 0.92 for pain, stiffness, and modified PF subscales respectively), construct validity could not be demonstrated. Factor analysis of the modified form of the WOMAC extracted four factors, which differed from the a priori triple stratification. However, factor analysis of the modified PF subscales extracted two factors, which accounted for 68.4% of the total variance and could be clinically characterized (disability during activities requiring knee flexion within the first 90 degrees and activities requiring knee flexion over more than 90 degrees ). CONCLUSION: We translated and adapted the WOMAC index into Arabic to suit Tunisian people. The translated questionnaire is reliable but not valid in its original form. We propose the use of a modified version of PF subscale of the WOMAC, although the psychometric properties of this instrument must be examined in a larger population. PMID- 15135143 TI - Osteoarthritis of the knee--clinical assessments and inflammatory markers. AB - OBJECTIVE: The present cross sectional study was performed to test the hypothesis that in osteoarthritis (OA) of the knee severity of this disease is related to local levels of inflammatory metabolites and their corresponding enzymes. METHODS: From 41 patients with OA of the knee (age range 45-79 years) undergoing arthroscopy blood, synovial fluid (SF) and synovial membrane (SM) were collected. Clinical conditions were primarily assessed by the WOMAC-index and radiographic grading (K&L-grade). Concentrations of PGE(2), TxB(2)and NO(2/3)and that of IL-6, IL-1 alpha, IL-1 beta, TNF alpha, COX-2 and iNOS were determined in SF and SM, respectively. RESULTS: With advancing age K&L-grade and COX-2 in SM increased significantly (P=0.005 and P=0.01, respectively). TNF alpha and IL-1 alpha were not detectable in SM samples. Apart from a correlation between PGE(2)and WOMAC index (r=0.36, P=0.035) no significant relationships could be found between the various inflammatory parameters and any of the assessed clinical signs. CONCLUSIONS: Apparently no direct relationships exist between the measured markers of inflammation (e.g. PGE(2), NO(2/3)) or the involved enzymes (e.g. COX 2, iNOS) and the severity of OA of the knee. The degenerative condition of this disease might be due to the more local, mainly mechanical injury with little systemic upset. However, further longitudinal studies are needed to clarify whether the assessed biochemical markers could serve as predictors for the progression of OA. PMID- 15135144 TI - Age related changes in human articular chondrocyte yield, proliferation and post expansion chondrogenic capacity. AB - OBJECTIVE: We investigated how aging effects human chondrocyte yield, proliferation, post-expansion chondrogenic capacity, and response to specific growth factors supplemented during expansion. METHODS: Fifty-two samples of human articular cartilage were harvested from cadavers 20 to 91 years old and grouped into age decades. Cell yields were normalised to tissue wet weight. Cell proliferation rates were calculated during expansion in medium without (CTR) or with TGF beta 1, FGF-2 and PDGF-BB (TFP). Chondrogenic capacity of CTR- and TFP expanded cells was assessed by cultivation as 3D pellets in a defined serum-free medium, followed by histological, immunohistochemical, biochemical and real-time RT-PCR analyses. RESULTS: Cell yields were similar in donors up to 40 years of age and significantly lower (1.8-fold) in older donors. Cell proliferation rates in CTR medium significantly decreased after 30 years of age and remained similar in older donors. In the presence of TFP, proliferation rates were higher (up to 3.7-fold) in all age groups and decreased only slightly with age. The glycosaminoglycan (GAG) content of pellets obtained from CTR-expanded cells was not correlated with age. Pellets from TFP-expanded cells reproducibly contained more GAG (up to 3.2-fold) than those from CTR-expanded cells only if donors were younger than 40. Safranin O staining intensity and collagen type II expression and accumulation were consistent with GAG contents. CONCLUSION: Medium supplementation with the growth factor combination TFP during chondrocyte expansion supports higher proliferation rates at any age and higher post expansion chondrogenic capacity in donors up to 40 years. These findings may be relevant for chondrocyte-based cartilage repair procedures. PMID- 15135145 TI - Increased stromelysin-1 (MMP-3), proteoglycan degradation (3B3- and 7D4) and collagen damage in cyclically load-injured articular cartilage. AB - OBJECTIVE: To determine whether load-induced injury causes alterations in proteoglycan (PG), stromelysin-1 (MMP-3) and collagen in articular cartilage. METHODS: Mature bovine cartilage was cyclically loaded at 0.5 Hz with 1 and 5 MPa for 1, 6 and 24h. Immediately after loading explants were evaluated for cell viability. Alterations in matrix integrity were determined by measuring PG content, PG degradation using 7D4 and 3B3(-) antibodies, broken collagen using COL2-3/4m antibody, and stromelysin-1 content using a MMP-3 antibody. RESULTS: Mechanical load caused cell death and PG loss starting from the articular surface and increasing in depth with loading time. There was a decrease in the 7D4 epitope (native chondroitin sulfate) in the superficial zone of cartilage loaded for longer than 1h, but an increase around chondrocytes in the deep zone. The 3B3(-) staining for degraded/abnormal chondroitin-4-sulfate neoepitope appeared only in cartilage loaded under the most severe condition (5 MPa, 24 h). The elevation of stromelysin-1 was co-localized with broken collagen (COL2-3/4m) at the articular surface in explants loaded with 1 and 5 MPa for 24 h. CONCLUSIONS: Cell death and PG loss occurred within 6h of cyclic loading. The elevation of MMP 3 following cell death was consistently found in the superficial zone of loaded cartilage. Since MMP-3 can degrade PG and super-activate procollagenase, the increase of MMP-3 can therefore induce matrix degradation and PG depletion in mechanically injured articular cartilage, both of which are important to the development of osteoarthritis. PMID- 15135147 TI - Effect of glucosamine supplementation on fasting and non-fasting plasma glucose and serum insulin concentrations in healthy individuals. AB - OBJECTIVE: To test the hypothesis that glucose intolerance does not occur when healthy adults consume normal, recommended dosages of glucosamine sulfate. METHODS: Healthy adults (N=19) ingested 1500 mg of glucosamine sulfate or placebo (double blind) each day for 12 weeks. Three-hour oral glucose tolerance tests (OGTT) were performed using 75 g of dextrose. These occurred before the start of supplementation, at 6 weeks, and at the completion of supplementation (12 weeks). RESULTS: There were no significant differences between fasted levels of serum insulin or blood glucose. Glucosamine sulfate supplementation did not alter serum insulin or plasma glucose during the OGTT. There were no significant differences within or between treatments, ages or gender. Glycated hemoglobin measurements at the three time points showed no significant change over time, within or between treatments, ages or gender. The lack of significant changes may have been due to large standard deviations in the data. CONCLUSION: The data suggests that glucosamine supplementation, with normal recommended dosages, does not cause glucose intolerance in healthy adults. This cannot be determined conclusively, however, until further studies are conducted using alternative types of testing. PMID- 15135146 TI - Demineralized bone alters expression of Wnt network components during chondroinduction of post-natal fibroblasts. AB - OBJECTIVE: The Wnt family of secreted proteins, their receptors (Fzd proteins) and antagonists (secreted Fzd-related proteins, or Sfrp) regulate chondrocyte differentiation and chrondrogenesis during embryonic development. Here, the hypothesis that the Wnt regulatory network contributes to chondrocyte differentiation of post-natal cells was tested in an in vitro model of chondroinduction by demineralized bone powder (DBP). DESIGN: Human dermal fibroblasts (hDFs) were cultured in porous, three-dimensional (3D) collagen sponges with or without chondroinductive DBP. In some experiments, lithium chloride (LiCl), an agonist of the Wnt/beta-catenin signaling pathway, was added to the culture media. Sponges were cultured for intervals (0.5-21 days) before processing for molecular, histologic, and biochemical analyses. Expression of wnt, fzd, and sfrp genes was characterized by semi-quantitative RT-PCR. Fibroblasts' contacts with DBP were documented by histology. Accumulation of proteoglycan in extracellular matrix was evaluated by histology (metachromasia in toluidine blue-stained sections) and quantitative immunoassay (chondroitin 4 sulfate ELISA). RESULTS: Expression of 15 wnt, fzd, and sfrp family members was detected in hDFs by RT-PCR. A subset of those genes (wnt2b, wnt5b, wnt10b, fzd6, fzd7) showed altered expression in hDFs exposed to DBP for 3 days. wnt and fzd gene expression was not altered before hDFs contacted the DBP within the collagen sponge. Human DFs cultured in plain collagen sponges and treated with LiCl accumulated significantly more metachromatic matrix than NaCl-treated controls on day 10, and showed a trend towards increased matrix chondroitin-4 sulfate content. CONCLUSIONS: These data suggest that changes in Wnt signaling contribute to chondroinduction of post-natal fibroblasts by DBP. This is the first evidence that Wnt components, which are essential regulators of pre-natal chondrocyte differentiation, may also influence post-natal chondrocyte differentiation induced by DBP. PMID- 15135148 TI - Cellular mechanisms of vitamin E uptake: relevance in alpha-tocopherol metabolism and potential implications for disease. AB - alpha-Tocopherol is an essential micronutrient involved in various oxidative stress-related processes. Because of its hydrophobic nature, alpha-tocopherol is transported in plasma lipoproteins, and the pathways involved in its cellular uptake are closely related to the lipoprotein metabolism. alpha-Tocopherol transfer from plasma to cells can occur by different mechanisms such as uptake facilitated by lipid transfer proteins and lipases, receptor-mediated lipoprotein endocytosis, and selective lipid uptake. Here we discuss recent progress in understanding the physiological and pathophysiological relevance of these different pathways for cellular uptake of vitamin E in vivo. This review is mainly focused on the role of the scavenger receptor class B type I (SR-BI) on alpha-tocopherol metabolism and its potential implications for disease conditions. PMID- 15135149 TI - Ethylenediaminetetraacetic acid (EDTA) does not increase iron uptake or ferritin synthesis by Caco-2 cells. AB - Ethylenediaminetetraacetic acid (NaFe-EDTA) is a chelator capable of binding a wide variety of metals, with a high affinity constant for Fe(3+). NaFe-EDTA has been extensively studied and validated as an excellent choice for iron fortification programs and extensive research has demonstrated its high bioavailability specially for cereal based foods. To further evaluate the usefulness of this compound we performed iron uptake experiments with EDTA using the Caco-2 cell system. Cells were incubated in PBS at pH 5.5 or 7.0, containing or not ascorbic acid. Different sources of EDTA, different concentrations of NaFe EDTA and the inclusion of another iron compound as electrolytic iron, were tested. Also, the ferritin content of Caco-2 cells 24h after 1h incubation with iron compounds was evaluated. Except for the addition of ascorbic acid, under the experimental conditions used, Caco-2 cells were not capable of obtaining iron from NaFe-EDTA. Furthermore, iron uptake from electrolytic iron was inhibited when Na(2) or K(2)-EDTA were included. Ferritin determinations to Caco-2 cells evaluated 24h after 1h incubation periods, showed that NaFe-EDTA did not induce new ferritin synthesis, since iron did not enter the cells. Further studies are required to evaluate incorporation of iron from NaFe-EDTA to a common iron pool and the requirements for iron uptake by Caco-2 cells. PMID- 15135150 TI - Antihypertensive effects of Undaria pinnatifida (wakame) peptide on blood pressure in spontaneously hypertensive rats. AB - We examined the angiotensin I-converting enzyme (ACE) inhibitory activity and antihypertensive effect of the hot water extract of wakame, Undaria pinnatifida. Ten dipeptides were isolated from the extract by several steps of chromatography, and their amino acid sequences were Tyr-His, Lys-Trp, Lys-Tyr, Lys-Phe, Phe-Tyr, Val-Trp, Val-Phe, Ile-Tyr, Ile-Trp, and Val-Tyr. Both single administration and repeated oral administration of synthetic Tyr-His, Lys-Tyr, Phe-Tyr, and Ile-Tyr significantly decreased blood pressure in spontaneously hypertensive rats. PMID- 15135151 TI - Dietary polyunsaturated fatty acids suppress acute hepatitis, alter gene expression and prolong survival of female Long-Evans Cinnamon rats, a model of Wilson disease. AB - In the Long-Evans Cinnamon rat, copper accumulates in the liver because of a mutation in the copper-transporting ATPase gene, and peroxidative stresses are supposed to be augmented. We examined the effects of dietary fatty acids on hepatitis, hepatic gene expression, and survival. Rats were fed a conventional, low-fat diet (CE2), a CE2 diet supplemented with 10 wt% of lard (Lar), high linoleic soybean oil (Soy), or a mixture of docosahexaenoic acid (DHA)-rich fish oil and soybean oil (DHA/Soy). Among female rats, the mean survival times of the DHA/Soy and the Soy groups were longer by 17 approximately 20% than in the Lar and the CE2 groups. Among male rats, the survival times were much longer than in the females, but no significant difference in survival was observed among the dietary groups. Serum ceruloplasmin levels in female and male rats of all of the dietary groups were similar. Serum transaminase levels of the DHA/Soy group tended to be lower than in the CE2 group. Histological examinations revealed a marked degeneration in hepatic tissue integrity in the Lar and CE2 groups but not in the DHA/Soy group. Hepatic levels of metal-related genes, transferrin and ceruloplasmin, as well as those related to bile acid synthesis were up-regulated, and an inflammation-related gene (cyclooxygenase [COX]-2) was down-regulated in the DHA/Soy group. Some proliferation-related genes were also affected by the dietary fatty acids. These results indicate that polyunsaturated fatty acids suppress the development of acute hepatitis and prolong survival in females, regardless of whether they are of the n-6 or n-3 type, which are associated with altered gene expressions. PMID- 15135152 TI - Metabolic effects of acute measles in chronically malnourished Nigerian children. AB - We hypothesized that acute measles infection imposes severe metabolic demands on malnourished children. Nigerian rural communities, characterized by severe poverty and extensive malnutrition, served as site for this study. Sixty-five children (mean [+/-SD] age 2.67 +/- 1.96 years) with measles and a randomly selected equal number of children (age 2.83 +/- 1.23 years) from the same communities but measles-free were studied. Both groups were serologically negative for human immunodeficiency virus. The percentages of nonmeasles group who were underweight and wasted as exemplified by weight for age (WAZ) and weight for height (WHZ) scores less than -2.0 SD were 43% and 23%, respectively. Comparative values for the measles group (66% and 54% respectively) were significantly (P < 0.01 or 0.001) different. Compared to the controls, measles infected children had significantly (P < 0.001) higher plasma cortisol level, marked hyporetinemia (plasma retinol 0.62 +/- 0.24 micromol/L) and prominent reduction (P < 0.002) in the sum of serum essential amino acids. Measles promoted a TH(1) to TH(2) cytokine shift, with severe depletion of plasma interleukin (IL) 12, a key cytokine in the development of cell mediated immunity. IL-6, a key stimulator of hepatic acute phase protein response, was prominently (P < 0.002) increased in plasma in measles-infected children. Glucocorticoids exert effects on cytokine expression, as well as on cytokine receptor expression and cytokine regulated biological responses. They enhance synergistically, the effects of IL-1 and IL-6 type cytokines on many acute phase proteins. Because of the prominent increase in circulating level of cortisol in acute measles, glucocorticoid treatment for associated sepsis may pose serious problems. Additionally, glucocorticoids antagonize several effects of retinoids at cellular and transcriptional levels, thus suggesting that hypercortisolemia may increase the requirement for retinoids. PMID- 15135153 TI - Oral absorption of phytosterols and emulsified phytosterols by Sprague-Dawley rats. AB - Clinical studies have demonstrated that consumption of phytosterol esters in lipid-based foods decreases serum concentrations of total and LDL cholesterol. These substances represent minimal potential for adverse effects when consumed orally because of their low bioavailability. However, some studies have reported estrogenic and other effects in laboratory animals treated parenterally with phytosterols, demonstrating that these substances may have the potential to cause adverse effects if absorbed. Water-soluble phytosterols have been prepared by formulation with emulsifiers to expand delivery options to include non-lipid based foods. However, emulsifiers are used as excipients in the formulation of lipophilic pharmaceuticals to increase solubility, thereby increasing their absorption. Therefore, oral consumption of emulsified water-soluble phytosterols could potentially increase their absorption. In the current study, absorption of phytosterols prepared as water-soluble emulsified micelles with two different food-grade emulsifiers was evaluated in Sprague-Dawley rats and compared with absorption of non-micellar free phytosterols and esterified phytosterol mixtures dissolved in a lipophilic vehicle (soybean oil). Rats were dosed via gavage with 42 mg/kg of formulated phytosterol preparations. Blood was collected at 8, 16, 24, and 32 hours, extracted with hexane, derivatized with benzoyl chloride, and analyzed by high-performance liquid chromatography to determine concentrations of beta-sitosterol, and campesterol. Plasma concentrations and AUC(0-32 hours) [microg/mL/h] of beta-sitosterol and campesterol were lower in plasma obtained from rats treated with emulsified phytosterol preparations than in animals treated with free phytosterols dissolved in soybean oil. Because the pharmacokinetic profile of water-soluble phytosterols is similar to that of phytosterols administered in a lipid vehicle, the safety profile is likely to be the same as that of phytosterols and phytosterol esters in currently used applications. PMID- 15135154 TI - Dietary fiber-rich colloids from apple pomace extraction juices do not affect food intake and blood serum lipid levels, but enhance fecal excretion of steroids in rats. AB - The aim of this study was to investigate the effects of colloids isolated from apple pomace extraction juices (so-called B-juices) produced by enzymatic liquefaction on food intake, levels of blood serum lipids, and fecal excretion of bile acids (BA) and neutral sterols (NS) in vivo. Ten male Wistar rats per group were fed diets containing either no apple dietary fiber (DF) (control), a 5% supplementation with juice colloids, or an alcohol-insoluble substance (AIS) from apples for 6 weeks. Apple DF in diets led to lower weight gain in rats fed with B juice colloids (P< 0.05). For these rats, food intake was not affected but was highest with feeding AIS (10% more than control) to cover energy requirements. The supplementation of diet with apple DF from extraction juices or AIS had minor effects on blood serum lipids. In rats fed either juice colloids or AIS, up to 30% (5.31 micromol/g dry weight) and 88% (7.69 micromol/g dry weight) more primary BA were excreted in feces, respectively, as compared to that in the control group (4.10 micromol/g dry weight) (P < 0.05). In cecal contents, a 15% (juice colloids) to 37% (AIS) increase in primary BA was found. In contrast, concentrations of secondary BA were lower in feces of test groups (P < 0.05). Excretion of total BA and NS was higher in rats fed apple DF (P < 0.05). Our study is the first to prove that there are beneficial physiologic effects of apple DF isolated from pomace extraction juices produced by enzymatic liquefaction. These results may help to develop such innovative juice products that are rich in DF of fruit origin for diminishing the lack of DF intake. PMID- 15135155 TI - Dimethylaminopurine inhibits metabolic effects of insulin in primary adipocytes. AB - Dimethylaminopurine (DMAP) has previously been used as an inhibitor of phosphorylation in studies of meiotic events, and more recently to investigate TNFalpha signaling, because of its potential to inhibit activation of c-jun N terminal kinase (JNK). Here we have addressed the effects of DMAP on metabolic insulin responses in adipocytes and on intracellular insulin signaling molecules. At 100 micromol/L, DMAP completely inhibited the ability of insulin to counteract lipolysis in isolated adipocytes. Insulin-induced lipogenesis and glucose uptake was inhibited to a lesser degree in a concentration-dependent manner starting at 10 micromol/L DMAP. Insulin-induced tyrosine phosphorylation of the insulin receptor was not affected by DMAP. Insulin-induced activation of protein kinase B, a known mediator of insulin action, was not inhibited by 100 micromol/L, but to a low extent by 1 mmol/L DMAP in intact cells. This inhibition was not sufficient to affect activation of the downstream protein kinase B substrate phosphodiesterase 3B. The inhibition of activation of JNK as a possible mechanism whereby DMAP affects insulin-induced antilipolysis, lipogenesis, and glucose uptake, was investigated using the JNK inhibitor SP600125. At 100 micromol/L, SP600125 completely reversed the antilipolytic effect of insulin, as well as partially inhibited insulin-induced lipogenesis and glucose-uptake, indicating that JNK may be involved in mediating these actions of insulin. Inhibition of JNK by DMAP may therefore partly explain the negative impact of DMAP on insulin action in adipocytes. PMID- 15135156 TI - Volume reduction of the right anterior limb of the internal capsule in patients with schizotypal disorder. AB - We have previously reported bilateral volume reductions in the anterior limb of the internal capsule (ALIC) in patients with schizophrenia. The purpose of this study was to extend the volumetric measurements of ALIC to subjects with schizotypal features to explore the neurobiology underlying schizophrenia spectrum disorders in view of the fronto-thalamic connectivity. Three-dimensional magnetic resonance images were acquired from 24 patients with schizotypal disorder (ICD-10) and 47 healthy volunteers matched for age, gender, handedness, and parental education. Volumetric analyses of the ALIC and anterior parts of the caudate and lentiform nuclei were conducted using consecutive 1-mm thick coronal slices rostral to the anterior commissure. Compared with the comparison subjects, the schizotypal patients had significantly decreased volume in the right ALIC, but there was no significant group difference in the left ALIC volume. Volumes of the anterior part of the caudate or lentiform nucleus did not differ between groups. Volume deficit confined to the right ALIC suggests that limited involvement of the fronto-thalamic connectivity may have some relevance to the sparing of schizotypal patients from the development of overt psychosis. PMID- 15135157 TI - Functional magnetic resonance imaging in schizophrenia: cortical response to motor stimulation. AB - Previous functional magnetic resonance imaging (fMRI) studies suggest that motor system abnormalities are present in schizophrenia. However, these studies have often produced conflicting or ambiguous findings. The purpose of this study was to ascertain whether activation differences could be identified in stable schizophrenic patients on the basis of BOLD measures in two motor regions, the primary motor cortex, Brodmann area 4 (BA4) and the premotor and supplementary motor area, Brodmann area 6 (BA6). Twenty-one schizophrenic patients and 21 healthy control subjects were studied with BOLD fMRI methods during a sequential finger tapping task. Statistical parametric maps were generated for each subject, and anatomic regions were automatically defined using an anatomic atlas. Compared with controls, the schizophrenic patients showed a significant reduction in contralateral activation for both BA4 and BA6 (P<0.001), and in ipsilateral activation in BA4 (P=0.007) and BA6 (P=0.002). In healthy controls, the coactivation in the ipsilateral cortex is reduced in comparison with the contralateral cortex for right and left handed tasks. In BA4, this reduction is significant for right (P=0.007) and left (P=0.003) finger tapping. Similar results were obtained for BA6. Further analyses are necessary to evaluate the activation in other motor system regions. PMID- 15135158 TI - Abnormal brain response of chronic schizophrenia patients despite normal performance during a visual vigilance task. AB - Deficits of attention are common among individuals with schizophrenia (SZ) and are related both to genetic liability to the disorder and to functional outcome among patients. To explore the brain systems underlying these attentional abnormalities, we compared the response of nine patients with chronic SZ or schizoaffective disorder to that of 10 matched healthy individuals performing a simple visual vigilance task during functional magnetic resonance imaging. The two groups performed equivalently on the task. When the blood oxygen level dependent (BOLD) signal during identification of a target letter among similar looking letters was compared to the response during fixation trials, both groups showed multiple clusters of significant brain response in widespread cortical regions. Compared with healthy participants, SZ patients showed a diminished response in the inferior frontal cortex and an abnormally enhanced response in right postcentral gyrus, right medial temporal lobe and left cerebellum. The results suggest that abnormalities of functional brain response to attentional tasks can be observed among patients with SZ even when behavioral performance is unimpaired, and provide further evidence that brain systems related to attention are likely to be involved in the pathophysiology of the disorder. PMID- 15135159 TI - Functional neuroanatomical correlates of eye movements during rapid eye movement sleep in depressed patients. AB - In depressed patients, REM density, or the number of rapid eye movements (REMs) per minute of REM sleep, is a correlate of depression severity and clinical outcomes. We investigated the functional neuroanatomical correlates of average REM counts (RC), an automated analog of REM density, in depression. Thirteen medication-free depressed patients underwent all night polysomnography and positron emission tomography (PET) scans using [(18)F]fluoro-2-deoxy-d-glucose ([(18)F] FDG) during REM sleep. Regression analyses were conducted with Statistical Parametric Mapping (SPM-99). Average RC significantly and positively correlated with relative regional cerebral metabolic rate of glucose (rCMRglc) bilaterally in the striate cortex, the posterior parietal cortices, and in the medial and ventrolateral prefrontal cortices. Average RC were negatively correlated with rCMRglc in areas corresponding bilaterally to the lateral occipital cortex, cuneus, temporal cortices, and parahippocampal gyri. The areas where average RC was positively correlated with rCMRglc appear to constitute a diffuse cortical system involved in the regulation of emotion-induced arousal. The observed pattern of correlations suggests that average RC may be a marker of hypofrontality during REM sleep in depressed patients. PMID- 15135161 TI - Frontal lobe GABA levels in cocaine dependence: a two-dimensional, J-resolved magnetic resonance spectroscopy study. AB - Non-invasive measures of brain gamma-aminobutyric acid (GABA) concentrations may be especially useful in the identification of cocaine-related changes in brain chemistry that can be used to guide the development of future treatments for cocaine-dependent persons. This study assessed whether brain GABA levels in cocaine-dependent subjects with and without an alcohol disorder differ from GABA levels in healthy comparison subjects. Two-dimensional, proton magnetic resonance spectroscopy was used to determine GABA levels in the left prefrontal lobe of cocaine-dependent subjects (N=35) recruited from a National Institute on Drug Abuse (NIDA)-sponsored treatment trial of cocaine dependence and a comparison group (N=20). At treatment baseline, mean GABA concentrations were 0.93+/-0.27 mM/kg in cocaine-dependent subjects and 1.32+/-0.44 mM/kg in the comparison sample (t [d.f.=53]=3.65, P<0.001). Cocaine-dependent subjects with a history of a co-morbid alcohol disorder (N=23) had significantly lower baseline GABA levels (0.87 mM/kg) (t [d.f.=41]=4.31, P<0.001) than the comparison group. However, cocaine-dependent subjects without an alcohol disorder (N=12) also had lower GABA levels (1.04 mM/kg) than the comparison subjects (t [d.f.=30]=2.09, P=0.045), suggesting that cocaine dependence alone can decrease GABA levels. PMID- 15135160 TI - Attenuation of cue-induced cigarette craving and anterior cingulate cortex activation in bupropion-treated smokers: a preliminary study. AB - In untreated smokers, exposure to cigarette-related cues increases both the intensity of cigarette craving and relative glucose metabolism of the perigenual/ventral anterior cingulate cortex (ACC). Given that treatment with bupropion HCl reduces overall cigarette craving levels in nicotine dependent subjects, we performed a preliminary study of smokers to determine if bupropion HCl treatment attenuates cue-induced cigarette craving and associated brain metabolic activation. Thirty-seven, otherwise healthy smokers (20 untreated and 17 who had received open-label treatment with bupropion HCl) underwent two (18)F fluorodeoxyglucose positron emission tomography scanning sessions in randomized order--one when presented with neutral cues and the other when presented with cigarette-related cues. Bupropion-treated smokers had smaller cigarette cue induced increases in craving scores on the Urge to Smoke (UTS) Scale and less activation of perigenual/ventral ACC metabolism from the neutral to the cigarette cue scan than untreated smokers. Thus, in addition to its known effects on spontaneous cigarette craving and withdrawal symptoms, bupropion HCl diminishes cue-induced cigarette craving and appears to attenuate cigarette cue-induced ACC activation. These results are consistent with the known effects of bupropion HCl, including its enhancement of catecholaminergic neurotransmission. PMID- 15135162 TI - The effect of CGX-1007 and CI-1041, novel NMDA receptor antagonists, on kindling acquisition and expression. AB - CGX-1007, a 17-amino acid polypeptide isolated from the venom of Conus geographus, is a novel NMDA receptor antagonist that is selective for the NR2B subunit. CI-1041 (PD 196860; Co 200461) is a novel, orally available NR2B selective antagonist. Both compounds possess anticonvulsant activity in a variety of well-established animal seizure models. The present study was designed to assess the effects of CGX-1007 and CI-1041 on the acquisition and expression of kindled seizures. In the corneal kindled rat, CGX-1007 [Epilepsia 36 (1998) 39] and CI-1041, administered p.o., 2h prior to the kindling stimulation displayed time- and dose-dependent block of fully expressed corneal kindled seizures (ED50 = 300 pmol and 2.5mg/kg for CGX-1007 and CI-1041, respectively). In amygdala kindled rats, acute treatment with CGX-1007 blocked the secondarily generalized kindled seizure in a dose-dependent manner. Complete protection against the secondarily generalized seizure was only observed at a dose that produced behavioral impairment (4 nmol). Acute treatment with CI-1041 did not provide any notable protection against secondarily generalized seizures. Neither compound provided protection against the focal kindled seizure. Chronic i.c.v. infusion of CGX-1007 or chronic oral administration of CI-1041 did not delay the acquisition of amygdala kindling. The results from these studies suggest that NMDA receptors containing the NR2B subunit may contribute to the expression of fully kindled secondarily generalized seizures; however, they appear less important for the development of kindling. The differential results obtained with CGX-1007 and CI 1041 suggest that several classes of mechanistically distinct NR2B antagonists may exist and that CGX-1007 may be less specific as a NR2B receptor antagonist than initially reported. PMID- 15135163 TI - The effect of CGX-1007 and CI-1041, novel NMDA receptor antagonists, on NMDA receptor-mediated EPSCs. AB - The N-methyl-D-aspartate (NMDA) receptor-gated ion channel is comprised of at least one NR1 subunit and any of four NR2 subunits (NR2A-D). The NR2 subunit confers different pharmacological and kinetic properties to the receptor. CGX 1007 (Conantokin G), a 17-amino acid polypeptide isolated from the venom of Conus geographus, is a novel NMDA receptor antagonist that is thought to be selective for the NR2B subunit. CGX-1007 has been reported to have highly potent, broad spectrum anticonvulsant activity in animal seizure models. CI-1041 is an investigational compound, which also possesses anticonvulsant activity and has been shown to be highly selective for NR2B containing NMDA receptors. Although both CI-1041 and CGX-1007 are reportedly NR2B specific antagonists, they differ in their ability to block amygdala-kindled seizures, suggesting that the mechanism of action of these compounds differs. The present study was designed to test the hypothesis that CI-1041 and CGX-1007 would differentially modulate the function of NMDA receptors at excitatory synapses. Using the whole cell patch clamp technique, CGX-1007 and CI-1041 were found to block CA1 pyramidal cell, NMDA receptor-mediated excitatory postsynaptic currents (N-EPSCs) in a concentration-dependent manner in hippocampal slices from P4-P6 animals. In contrast, only CGX-1007 decreased NMDA receptor-mediated EPSC peak amplitude in slices from adult animals. The CGX-1007 block of peak amplitude was accompanied by a similar concentration-dependent decrease in decay kinetics of NMDA receptor mediated EPSCs. These results suggest that while CI-1041 may be selective for NMDA receptors containing the NR2B subunit, CGX-1007 appears to be less selective than previously reported. PMID- 15135164 TI - MAO(A) knockout mice are more susceptible to seizures but show reduced epileptogenesis. AB - The role of elevated neuroactive amine exposure during embryonic and early postnatal development on seizure threshold and epileptogenesis was examined using both electrical and pentylenetetrazol (PTZ) kindling in monoamine oxidase A knockout (MAO(A) KO) mice and their wildtype, parental strain (C3H). In the first experiment permanent bilateral electrodes were implanted in the amygdala of both C3H and MAO(A) KO mice. The mice had their afterdischarge threshold determined and then seizures were kindled daily for a total of 20 days. We observed that the MAO(A) KO mice had lower afterdischarge thresholds and less severe seizures compared to the C3H mice. In the second experiment, seizures were elicited in experimentally naive mice using 50mg/kg of PTZ once daily for 7 days. We observed that the MAO(A) KO mice had shorter latencies to the onset of the first seizure, shorter total duration of seizures and fewer seizures per day. Overall the results of both experiments suggest that MAO(A) KO mice have an increased susceptibility to seizures, but are more resistant to epileptogenesis. We conclude that the high levels of neuroactive amines in the MAO(A) KO mice reorganize the brain to make the mice more susceptible to seizures but the remaining high levels of serotonin and norepinephrine likely inhibit epileptogenesis. PMID- 15135165 TI - Levetiracetam during 1-year follow-up in children, adolescents, and young adults with refractory epilepsy. AB - PURPOSE: To evaluate the efficacy and safety of levetiracetam (LEV) in refractory crypto/symptomatic, partial or generalised epilepsy in children, adolescents and young adults. METHODS: We performed a prospective open label add-on study in 99 patients (age 12 months to 32 years, mean 14 years) with partial or generalised, crypto/symptomatic seizures. Levetiracetam was added to no more than two baseline AEDs and the efficacy was rated according to seizure type and frequency. RESULTS: LEV was initiated at the starting dose of 10mg/kg/day with 5-day increments up to 50 mg/kg/day, unless it was not tolerated. Concomitant therapy was generally not modified throughout the study. After a mean follow-up period of 6.7 months (range 3 weeks to 29 months), 11 patients (11.1%) were free of seizures (cryptogenic partial epilepsy, 5; symptomatic partial epilepsy, 6). A more than 75% seizure decrease was found in 14 patients (14.1%) and >50% in 8 (8.1%). Seizures were unchanged in 38 (38.4%), and worsened in 23 (23.2%). Mild and transient adverse side effects were found in 17 patients (17.2%), mostly represented by irritability and drowsiness. CONCLUSION: LEV appears to be well tolerated in children and adolescents with severe epilepsy and seems to be a broad spectrum AED, though in our experience, it was more effective against partial seizures with or without secondarily generalisation. LEV efficacy in other epilepsy syndrome should be evaluated further in homogeneous, more selected patients. PMID- 15135167 TI - Cardiorespiratory findings in sudden unexplained/unexpected death in epilepsy (SUDEP). AB - Sudden unexplained/unexpected death in epilepsy (SUDEP), with an incidence of 0.35-9.3/1000 patient-years depending on the severity of epilepsy, remains a diagnostic and therapeutic challenge. Potential pathomechanisms comprise cardiac arrhythmia, due to myocardial ischemia, electrolyte disturbances, arrhythmogenic drugs, or transmission of the epileptic activity via the autonomic nervous system to the heart, and central or obstructive apnea. In most studies on SUDEP, data are lacking about the family and patient's own clinical history, cardiovascular symptoms, concomitant diseases and prior findings. Whether arterial hypertension, diabetes, hypercholesterolemia, other neurologic disorders, lung diseases, smoking or electrolyte disturbances are risk factors for SUDEP is unknown. Whereas cardiac dysfunction during seizures has been documented by electrocardiography, and cardiac abnormalities are found in up to 33% of SUDEP cases autoptically, investigations between seizures found only little cardiac abnormalities. More knowledge about the cardiovascular and pulmonary status of epileptic patients during, immediately after and between seizures is needed, which may contribute to better understand and possibly prevent SUDEP by measures like "cardioprotective" drugs, respiratory therapy or implantation of a cardioverter/defibrillator. PMID- 15135166 TI - A comparison of three NMDA receptor antagonists in the treatment of prolonged status epilepticus. AB - Three different classes of NMDA receptor antagonists were compared for their effectiveness in terminating prolonged status epilepticus (SE), induced by continuous hippocampal stimulation. Animals were treated after 150 min of SE by intraperitoneal administration of increasing doses of 3-((R,S)-2-carboxypiperazin 4-yl)-propyl-1-phosphonic acid (CPP), MK-801 (dizocilpine), ifenprodil, or saline. EEG recordings were used to determine seizure termination. The first experiment (n = 57 animals) determined the most effective anticonvulsant dose of each agent by determining its ability to terminate SE within the next 300 min. Five control rats treated with normal saline after 150 min of SE continued to exhibit continuous seizures for the next 300 min. All drugs were administered after 150 min of SE. CPP terminated seizures with an ED(50) of 6.4 mg/kg; the maximal effective dose was 15 mg/kg. MK-801 has an ED(50) of 1.4 mg/kg; the maximal effective dose was 2 mg/kg. Ifenprodil was maximally effective at 30 mg/kg. However, an ED(50) could not be calculated. In a subsequent experiment, the NMDA antagonists were compared for their ability to terminate prolonged SE within 60 min of their administration at the most effective dose. MK-801 (2.0 mg/kg) terminated SE in 6 of 10 animals within 60 min, CPP (15 mg/kg) terminated it in 1 of 9 animals; ifenprodil (30 mg/kg) did not terminate it in any of 9 animals treated. In the 300 min following administration, CPP (6/9) and MK-801 (6/10) were equally efficacious in terminating SE but ifenprodil (2/7) was less effective (P = 0.065, chi-square test). The results indicate that the non competitive NMDA receptor antagonist MK-801 was superior to the competitive antagonist CPP and the pH-sensitive site antagonist ifenprodil, in terminating prolonged experimental SE. PMID- 15135168 TI - Hyperoxia is not an essential condition for status epilepticus induced cell death in organotypic hippocampal slice cultures. AB - The low Mg2+ model of epilepsy in organotypic hippocampal slice cultures is used to elucidate the mechanism underlying neuronal cell death following sustained epileptiform activity. However, the high oxygen tension of 95% widely used in this model is capable of inducing neuronal cell death by itself. Here we demonstrate that even under normoxic conditions 1h of epileptiform activity induced neuronal cell death as assessed by Propidium Iodide uptake. We conclude that hyperoxia is not essential for status epilepticus induced neuronal cell death in this model. PMID- 15135169 TI - Neurosyphilis and status epilepticus: case report and literature review. AB - We review the literature concerning the initial presentation of neurosyphilis in status epilepticus (SE) and provide an additional case of a 41-year-old male with no past medical history of seizures who presented in status epilepticus with subsequent laboratory confirmation of neurosyphilis. Neurosyphilis should be considered in the differential diagnosis of patients presenting with status epilepticus. PMID- 15135170 TI - Probing the free radicals formed in the metmyoglobin-hydrogen peroxide reaction. AB - The reaction between metmyoglobin and hydrogen peroxide results in the two electron reduction of H2O2 by the protein, with concomitant formation of a ferryl oxo heme and a protein-centered free radical. Sperm whale metmyoglobin, which contains three tyrosine residues (Tyr-103, Tyr-146, and Tyr-151) and two tryptophan residues (Trp-7 and Trp-14), forms a tryptophanyl radical at residue 14 that reacts with O2 to form a peroxyl radical and also forms distinct tyrosyl radicals at Tyr-103 and Tyr-151. Horse metmyoglobin, which lacks Tyr-151 of the sperm whale protein, forms an oxygen-reactive tryptophanyl radical and also a phenoxyl radical at Tyr-103. Human metmyoglobin, in addition to the tyrosine and tryptophan radicals formed on horse metmyoglobin, also forms a Cys-110-centered thiyl radical that can also form a peroxyl radical. The tryptophanyl radicals react both with molecular oxygen and with the spin trap 3,5-dibromo-4 nitrosobenzenesulfonic acid (DBNBS). The spin trap 5,5-dimethyl-1-pyrroline N oxide (DMPO) traps the Tyr-103 radicals and the Cys-110 thiyl radical of human myoglobin, and 2-methyl-2-nitrosopropane (MNP) traps all of the tyrosyl radicals. When excess H2O2 is used, DBNBS traps only a tyrosyl radical on horse myoglobin, but the detection of peroxyl radicals and the loss of tryptophan fluorescence support tryptophan oxidation under those conditions. Kinetic analysis of the formation of the various free radicals suggests that tryptophanyl radical and tyrosyl radical formation are independent events, and that formation of the Cys 110 thiyl radical on human myoglobin occurs via oxidation of the thiol group by the Tyr-103 phenoxyl radical. Peptide mapping studies of the radical adducts and direct EPR studies at low temperature and room temperature support the conclusions of the EPR spin trapping studies. PMID- 15135171 TI - The carbonate radical anion-induced covalent aggregation of human copper, zinc superoxide dismutase, and alpha-synuclein: intermediacy of tryptophan- and tyrosine-derived oxidation products. AB - In this review, we describe the free radical mechanism of covalent aggregation of human copper, zinc superoxide dismutase (hSOD1). Bicarbonate anion (HCO3-) enhances the covalent aggregation of hSOD1 mediated by the SOD1 peroxidase dependent formation of carbonate radical anion (CO3*-), a potent and selective oxidant. This species presumably diffuses out the active site of hSOD1 and reacts with tryptophan residue located on the surface of hSOD1. The oxidative degradation of tryptophan to kynurenine and N-formyl kynurenine results in the covalent crosslinking and aggregation of hSOD1. Implications of oxidant-mediated aggregation of hSOD1 in the increased cytotoxicity of motor neurons in amyotrophic lateral sclerosis are discussed. PMID- 15135172 TI - Effects of cyclophosphamide and buthionine sulfoximine on ovarian glutathione and apoptosis. AB - Treatment with the anticancer drug cyclophosphamide (CPA) destroys ovarian follicles. The active metabolites of CPA are detoxified by conjugation with glutathione (GSH). We tested the hypotheses that CPA causes apoptosis in ovarian follicles and that suppression of ovarian GSH synthesis before CPA administration enhances CPA-induced apoptosis. Proestrous rats were given two injections, 2 h apart, with (1) saline, then saline; (2) saline, then 50 mg/kg CPA; (3) saline, then 300 mg/kg CPA; or (4) 5 mmol/kg buthionine sulfoximine (BSO) to inhibit glutamate cysteine ligase (GCL), the rate-limiting enzyme in GSH synthesis, and then 50 mg/kg CPA. Statistically significantly increased DNA fragmentation by agarose gel electrophoresis and granulosa cell apoptosis by TUNEL were observed in the CPA-treated ovaries 24 h after the second injection, but BSO did not enhance the effect of 50 mg/kg CPA. We next tested the hypothesis that CPA depresses ovarian GSH concentration and expression of the rate-limiting enzyme in GSH synthesis, GCL. Proestrous rats were injected with 300 or 50 mg/kg CPA or vehicle and were sacrificed 8 or 24 h later. After CPA treatment, ovarian and hepatic GSH levels decreased significantly, and ovarian GCL subunit mRNA levels increased significantly. There were no significant changes in GCL subunit protein levels. Finally, we tested the hypothesis that GSH depletion causes apoptosis in ovarian follicles. Proestrous or estrous rats were injected with 5 mmol/kg BSO or saline at 0700 and 1900 h. There was a significant increase in the percentage of histologically atretic follicles and a nonsignificant increase in the percentage of apoptotic, TUNEL-positive follicles 24 h after onset of BSO treatment. Our results demonstrate that CPA destroys ovarian follicles by inducing granulosa cell apoptosis and that CPA treatment causes a decline in ovarian GSH levels. More pronounced GSH suppression achieved after BSO treatment did not cause a statistically significant increase in follicular apoptosis. Thus, GSH depletion does not seem to be the mechanism by which CPA causes follicular apoptosis. PMID- 15135173 TI - Oligonucleotides in human urine do not contain 8-oxo-7,8-dihydrodeoxyguanosine. AB - The promutagenic DNA modification 8-oxo-7,8-dihydrodeoxyguanosine is the most frequently used marker for oxidative stress to DNA. The unmodified base and nucleoside and the 8-hydroxylated guanine base and nucleoside are found in urine, the latter used as a global measure of oxidative stress to DNA. Nucleotide excision repair (NER) excises a 27- to 29-mer oligonucleotide with oxidative lesions, and if found in urine, it could be used as a measure of DNA repair in vivo. Enzymatic hydrolysis of human urines followed by HPLC-tandem mass spectrometry was not able to reveal oligonucleotides and/or mononucleotides with the 8-oxo-7,8-dihydrodeoxyguanosine modification. The recovery of a synthetic oligonucleotide with the modification was complete (95% confidence limits: 98 124%). These experiments show that oligonucleotides are excreted into urine, but that 8-oxo-7,8-dihydrodeoxyguanosine is found only as the mononucleoside and is not present in any significant amounts in oligonucleotides. We conclude that oligonucleotides are excreted into urine, and they do not contain oxidized lesions. Either NER products are degraded after excision or NER functions differently in vivo in humans compared with cellular systems. PMID- 15135174 TI - Quantification of 1,N6-etheno-2'-deoxyadenosine in human urine by column switching LC/APCI-MS/MS. AB - 1,N6-etheno-2'-deoxyadenosine (epsilondA) is one of several promutagenic DNA modifications arising from cellular oxidative metabolism. It is believed that these background DNA lesions may contribute to various diseases, such as cancer. Therefore, human biomonitoring of epsilondA in urine could be a potential marker for oxidative stress-related DNA damage. Existing methods for quantifying urinary epsilondA use 32P postlabeling. We have developed a nonradioactive, fast, and easier method based on column-switching liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry (LC/APCI-MS/MS) in the positive mode. Differences in column temperatures were used to influence analyte retention and sample focusing. With multiple reaction monitoring (MRM) mode the afforded limit of detection was about 0.7 pM when starting with 3 ml of urine. The urinary excretion rates of epsilondA from 28 nonsmoking and 5 smoking men were 10.0-99.6 pmol/24 h, and did not correlate with body weight, age, or plasma vitamin C concentration. The 5 smokers excreted 30.5 +/-8.5 and the 28 nonsmokers excreted 38.6 +/- 2.4 pmol epsilondA per 24 h, p=.37 (mean +/- SEM). The demonstrated level of performance suggests the future applicability of this method to studies of cancer and other diseases related to oxidative stress in humans. PMID- 15135175 TI - Oxidative stress promotes tau dephosphorylation in neuronal cells: the roles of cdk5 and PP1. AB - Oxidative stress has been demonstrated to produce modifications in several intracellular proteins that lead to alterations in their activities. Alzheimer's disease is related to an increase of oxidative stress markers, which may be an early event in the progression of the disease and neurofibrillary tangles formation. Abnormal phosphorylation of tau has been implicated in the etiopathogenesis of Alzheimer's disease. By using phospho-specific antibodies, we analyzed the changes in tau phosphorylation patterns after treatment of rat hippocampal and SHSY5Y human neuroblastoma cells with H2O2. We found that tau isoforms were hypophosphorylated at the Tau1 epitope after 2 h in the presence of H2O2. The decrease in the phosphorylation levels of tau protein were prevented by pretreatment with N-acetyl-L-cysteine. These changes were shown to depend on the activity of the cdk5/p35 complex, since a 3-fold increase in substrate phosphorylation and a 2-fold increase for the complex association were observed. Also, a decrease in the amount of inhibitor-2 bound to phosphatase PP1 was found in SHSY5Y cells under oxidative stress conditions. This decrease of inhibitor-2 bound to PP1 is due to an increased phosphorylation of the inhibitor-2 protein, thus leading to increased PP1 activity. Therefore, we propose that oxidative stress-induced activation of cdk5 leads to inhibitor-2 phosphorylation, relieving its inhibitory effect on PP1. PMID- 15135176 TI - Oxygen free radical scavenger enzyme polymorphisms in systemic sclerosis. AB - We performed a case-control study of polymorphisms of glutathione S-transferase (GST) isoenzymes and manganese superoxide dismutase (MnSOD) in black South Africans with systemic sclerosis (SSc). The frequency of the GSTM1*B phenotype was significantly decreased in the overall SSc group compared with controls (OR=0.19, p(corr)<.05), implying a possible protective effect against development of the disease. There was also a trend toward increased MnSODAla allele and phenotype frequencies in the diffuse cutaneous SSc subset compared with controls (OR=2.11 and 3.15, respectively, p(corr)<.1). Our findings provide new data on the distribution of GST and MnSOD polymorphisms in healthy Africans and further evidence that genetic factors may have a contributory role to play in predisposing to oxidative stress in SSc. PMID- 15135177 TI - Effects of platinum and palladium ions on the production and reactivity of neutrophil-derived reactive oxygen species. AB - This study was designed to investigate the effects of platinum, as hydrogen hexachloroplatinate (Pt; 0.0025-25 microM), on the production of reactive oxygen species (ROS) by human neutrophils in vitro. ROS were measured by lucigenin enhanced chemiluminescence (LECL). Addition of Pt to neutrophils was accompanied by a lag phase of about 1 min, followed by a linear dose-related increase in LECL, which peaked at around 4 min and achieved statistical significance at concentrations of 0.025 microM Pt and higher. Interestingly, Pt-mediated enhancement of LECL was not associated with meaningful alterations in neutrophil oxygen consumption, assembly of NADPH oxidase, or cytosolic Ca2+ and was completely attenuated by superoxide dismutase and inhibitors of NADPH oxidase, but not by catalase or scavengers of hydroxyl radical, and was undetectable with cells from individuals with chronic granulomatous disease. Exposure of alpha1 proteinase inhibitor to Pt-treated neutrophils resulted in inactivation of elastase-inhibitory capacity, underscoring the potential toxicity of neutrophil/Pt interactions. The pro-oxidative actions of Pt were mimicked by palladium (Pd), but not by cisplatin or rhodium. These observations demonstrate that Pt and Pd potentiate the reactivity, as opposed to the generation of neutrophil-derived oxidants, an activity that may contribute to airway inflammation in occupationally and possibly environmentally exposed individuals. PMID- 15135178 TI - Redox ranking of inducers of a cancer-protective enzyme via the energy of their highest occupied molecular orbital. AB - Induction of phase 2 enzymes is a major strategy in chemoprotection against cancer. Inducers belong to nine different chemical classes. In this study we found that a measure of the tendency of 30 plant phenylpropenoids and synthetic analogs to release electrons correlates linearly with their potency in inducing the activity of NAD(P)H:quinone reductase (NQO1), a prototypic phase 2 cancer protective enzyme. The tendency to release electrons was determined by the energy of the highest occupied molecular orbital (E(HOMO)), calculated by simple quantum mechanical methods. The correlations observed establish a clear conclusion: the smaller the absolute E(HOMO) of an agent, A, i.e., the lower its reduction potential, E(A*+/A), the stronger is its electron donor property and the greater its inducer potency. The finding of this redox ranking of the inducers demonstrates the possibility of controlling and predicting the genetic expression of an enzymatic defense against cancer by xenobiotics via one physicochemical parameter, the reduction potential, E(A*+/A). PMID- 15135179 TI - Short-term vitamin E intake fails to improve cognitive or psychomotor performance of aged mice. AB - The purpose of this study was to determine if relatively short-term vitamin E supplementation could reverse age-associated impairments in cognitive or motor function and the accumulated oxidative damage in the brain of aged mice. Separate groups of 5- or 20-month-old C57BL6 mice were placed on either a control diet or the same diet supplemented with alpha-tocopheryl acetate (1.65 g/kg). After 4 weeks on the diets, mice were tested for cognitive and motor functions over the next 8 weeks, during which the supplementation was maintained. Vitamin E supplementation increased the concentration of alpha-tocopherol in the cerebral cortex of both the young and old mice, but did not significantly affect oxidative damage to proteins and lipids in the brain cortex. When compared with young controls, the old control mice showed slower learning of a swim maze, longer reaction times, diminished auditory and shock-startle responsiveness, and diminished motor performance on tests of coordinated running and bridge walking. The vitamin E-administered old mice failed to show improvement of function relative to age-matched controls on any of the tests, but did show altered retention performance on the swim maze task and impaired performance in the test of coordinated running. The latter effects were not evident in young mice on the supplemented diet. Results of this study suggest that, when implemented in relatively old mice, supplementation of vitamin E is ineffective in reversing preexisting age-related impairments of cognitive or motor function, and has little effect on common measures of protein or lipid oxidative damage in the mouse brain. Moreover, the current findings indicate that vitamin E could have detrimental effects on some brain functions when implemented in older animals. PMID- 15135180 TI - Cadmium inhibits the electron transfer chain and induces reactive oxygen species. AB - Recent research indicates that cadmium (Cd) induces oxidative damage in cells; however, the mechanism of the oxidative stress induced by this metal is unclear. We investigated the effects of Cd on the individual complexes of the electron transfer chain (ETC) and on the stimulation of reactive oxygen species (ROS) production in mitochondria. The activity of complexes II (succinate:ubiquinone oxidoreductase) and III (ubiquinol:cytochrome c oxidoreductase) of mitochondrial ETC from liver, brain, and heart showed greater inhibition by Cd than the other complexes. Cd stimulated ROS production in the mitochondria of all three tissues mentioned above. The effect of various electron donors (NADH, succinate, and 2,3 dimethoxy-5-methyl-6-decyl-1,4-benzoquinol) on ROS production was tested separately in the presence and in the absence of Cd. ESR showed that complex III might be the only site of ROS production induced by Cd. The results of kinetic studies and electron turnover experiments suggest that Cd may bind between semiubiquinone and cytochrome b566 of the Q0 site of cytochrome b of complex III, resulting in accumulation of semiubiquinones at the Q0 site. The semiubiquinones, being unstable, are prone to transfer one electron to molecular oxygen to form superoxide, providing a possible mechanism for Cd-induced generation of ROS in mitochondria. PMID- 15135181 TI - CO2 enhanced peroxidase activity of SOD1: the effects of pH. AB - At pH 7.4, CO2, rather than HCO3-, markedly enhances the oxidation of diverse substrates by SOD1 plus H2O2. Since the concentration of CO2 would fall with rising pH in HCO3- buffers, it was of interest to explore the effects of pH on the peroxidase activity of SOD1 in the presence and in the absence of HCO3-. The rate of NADPH peroxidation in the HCO3- buffer was minimally affected by pH in the range of 8-10.5; in a pyrophosphate buffer, the rate increased markedly, such that at pH 10.5 the rates in the two buffers were nearly identical. Similar results were obtained when urate was used as the peroxidizeable substrate. These results are explicable on the basis of an increase in the rate with pH due to the ionization of H2O2 to the effective HO2- coupled with a decrease in [CO2] due to the ionizations of H2CO3, which displaces the hydration equilibrium to the right. These two opposing effects counteract in the HCO3(-)-buffered reaction mixtures; in the pyrophosphate buffer, only the effect of increasing [H02-] was seen. PMID- 15135182 TI - Oxidation of pyocyanin, a cytotoxic product from Pseudomonas aeruginosa, by microperoxidase 11 and hydrogen peroxide. AB - Pyocyanin (1-hydroxy-N-methylphenazine) is a cytotoxic pigment secreted by the bacterial species Pseudomonas aeruginosa, which frequently infects the lungs of immunosuppressed patients as well as those with cystic fibrosis. Pyocyanin toxicity results presumably from the ability of the compound to undergo reduction by NAD(P)H and subsequent generation of superoxide and H2O2 directly in the lungs. We report that in the presence of peroxidase mimics, microperoxidase 11, or hemin, pyocyanin undergoes oxidation by H2O2, as evidenced by loss of the pigment's characteristic absorption spectrum and by EPR detection of a free radical metabolite. The oxidation of pyocyanin is irreversible, suggesting an extensive modification of the pigment's phenazine chromophore. Oxidation of pyocyanin was observed also when exogenous H2O2 was replaced by a H2O2-generating system consisting of NADH and the pigment itself. That the oxidation involves the phenolate group of pyocyanin was verified by the observation that a related pigment, phenazine methosulfate, which is devoid of this group, does not undergo oxidation by microperoxidase 11/H2O2. In contrast to intact pyocyanin, oxidized pyocyanin was less efficient in NADH oxidation and stimulation of interleukin-8 release by human alveolar epithelial A549 cells in vitro, suggesting that oxidation of pyocyanin leads to its inactivation. This study demonstrates that pyocyanin may play a dual role in biological systems, first as an oxidant and ROS generator, and second as a substrate for peroxidases, contributing to H2O2 removal. This latter property may cause pyocyanin degradation and inactivation, which may be of considerable biomedical interest. PMID- 15135183 TI - Mitochondrial metabolism underlies hyperoxic cell damage. AB - Exposure of mammals to hyperoxia causes pulmonary and ocular pathology. Hyperoxic damage and cell death may derive from enhanced intracellular formation of reactive oxygen species (ROS), probably of mitochondrial origin. There is, however, controversy on this point. When wild-type and respiration-deficient (rho(o)) HeLa cells were cultured in 80% O2, wild-type cells stopped growing after 5 days and died thereafter whereas rho(o) cells survived and grew to confluence. This tolerance of rho(o) cells for hyperoxia was not associated with greater resistance to oxidants such as hydrogen peroxide and t-butyl hydroperoxide. Under both 20% and 80% O2, rho(o) cells exhibited substantially decreased ROS production, and, under 80% O2, rho(o) cells showed no suppression of aconitase activity or mitochondrial protein carbonyl formation. Replacement of normal mitochondria in rho(o) cells restored ROS production and susceptibility to hyperoxia. Two other approaches that diminished mitochondrial ROS generation also increased tolerance for hyperoxia. HeLa cells constantly exposed to the protonophoric uncoupler carbonyl cyanide m-chlorophenylhydrazone, which enhances respiration but decreases ROS production, showed preferential survival under 80% O2, as did HeLa cells treated with chloramphenicol, which suppresses both respiration and mitochondrial ROS production. We conclude that interactions between respiring mitochondria and O2 are primarily responsible for hyperoxic cell damage. PMID- 15135184 TI - Homocysteine and Alzheimer's disease: a modifiable risk? AB - A hypothesis is proposed that reconciles the epidemiological observation of elevated homocysteine in Alzheimer's disease (AD) with clinical features of the disease, particularly evidence of increased oxidative stress. We propose homocysteine is involved in an iron dysregulation/oxidative stress cycle that has a central role in the pathogenesis of AD. The implications of the hypothesis and some strategies for testing it are discussed. PMID- 15135186 TI - Resuscitation Great. Sir Edward Sharpey-Schafer and his simple and efficient method of performing artificial respiration. PMID- 15135187 TI - Impact of attending a 1-day multi-professional course (ALERT) on the knowledge of acute care in trainee doctors. AB - We have described previously deficiencies in the knowledge in trainee doctors of aspects of acute illness, its recognition and management. This led to the development of a 1-day multi-professional course in acute care for newly qualified doctors and nurses, ALERT. Using a questionnaire, we assessed the knowledge of basic aspects of acute care amongst 118 senior house officers, 36 of whom had previously attended an ALERT course. The average (+/-S.D.) knowledge score was higher for those who had completed an ALERT course (9.44 +/- 1.63 points versus 7.45 +/- 2.32 points; P < 0.05). In addition, those in the post ALERT group also showed significantly better knowledge of the signs of complete airway obstruction, normal capillary refill time, percentage survival after in hospital cardiac arrest, consent arrangements for operation in unconscious patients, minimum hourly urine output, the need to inflate the reservoir bag on a high concentration oxygen mask and the role of the reservoir. Similar differences existed between trainees who had completed an ALERT course and a group of SHOs assessed in 1991, who had not done so. We believe that we have demonstrated evidence that doctors' knowledge of acute care can be improved by attending courses such as ALERT. PMID- 15135188 TI - Long-term survival and neurological outcome of patients who received recombinant tissue plasminogen activator during out-of-hospital cardiac arrest. AB - OBJECTIVE: The long-term outcome in patients who received recombinant tissue plasminogen activator during cardiopulmonary resuscitation (CPR) in out-of hospital cardiac arrest (OHCA) of non-traumatic aetiology was assessed. METHODS: The neurological outcome in survivors and their level of performance, subjective well-being and quality of life were evaluated. RESULTS: A follow-up study of 27 cardiac arrest survivors was conducted; four patients (15%) died during the first year, a total of seven patients (26%) within 5 years. Twenty-two patients (81%) were discharged from hospital without neurological deficit (cerebral performance category (CPC) score: 1), three patients scored CPC 2 and two patients CPC 3. Heart failure classification on discharge was, according to the New York Heart Association (NYHA) criteria 2.1 +/- 0.9. Fifteen patients (56%) managed to return to their previous level of activity. At the time of follow-up 18 patients (67%) were still alive, of whom 15 responded to a survey regarding life satisfaction. Thirteen patients (87%) judged their situation to be worth living and twelve (80%) considered their survival a second chance, while five (33%) feared they could suffer another cardiac arrest. Reactions from close relatives included fear/anxiety (n = 14; 78%), a sustained burden on family life (n = 12; 67%), and occasional depression (n = 7; 39%). CONCLUSIONS: Thrombolytic therapy during cardiopulmonary resuscitation may produce a favourable neurological outcome. The majority of long-term survivors reported a good subjective quality of life. In one-third of close family members some negative factors had a lasting impact on the quality of daily living. PMID- 15135189 TI - Feasibility of shock advice analysis during CPR through removal of CPR artefacts from the human ECG. AB - Mechanical activity from chest compressions and ventilations during cardiopulmonary resuscitation (CPR) introduces artefact components into the electrocardiogram (ECG). CPR must therefore be discontinued for reliable shock advice analysis in automated external defibrillators. Reducing or eliminating this detrimental "hands-off" time by removing the CPR artefacts, should significantly improve the defibrillation success rate. The feasibility of this was tested by removing the CPR artefacts using a multichannel adaptive filter, the multichannel recursive adaptive matching pursuit (MC-RAMP) algorithm. Human ECG and reference channel data from episodes with both shockable and non shockable underlying heart rhythms were recorded from 105 patients with out-of hospital cardiac arrest. The performance of a shock advice algorithm was evaluated before and after artefact removal using the MC-RAMP algorithm. From a test set consisting of 92 shockable and 174 non-shockable episodes a sensitivity of 96.7% and specificity of 79.9% was achieved, an increase of approximately 15 and 13%, respectively, compared to no filtering. Good sensitivity was achieved, enabling ECG analysis during CPR that would reduce the hands-off time on patients with shockable rhythms. However, CPR artefact removal on non-shockable rhythms proved a more difficult problem. We need a better understanding of the physiological mixing of artefacts and the underlying heart rhythm and suggest clinical trials to investigate the nature of CPR artefacts further. PMID- 15135190 TI - Adherence to guidelines when positioning the defibrillation electrodes. AB - BACKGROUND: Placement of the defibrillation electrodes affects the transmyocardial current and thus the success of a defibrillation attempt. In the international guidelines 2000, the placement of the apical electrode was changed more laterally to the mid-axillary line. Finnish national guidelines, based on the international guidelines, were published in 2002. OBJECTIVES: The purpose of this study was to determine to what extent health care professionals adhere to the new guidelines when positioning the electrodes. METHODS: Professionals were recruited from emergency medical services, university hospitals and primary care. Not revealing the purpose of the test, participants were asked to place self adhesive electrodes on a manikin as they would do in the resuscitation situation and to answer a questionnaire about resuscitation training and familiarity with the guidelines. The distance of electrodes from the recommended position was measured in horizontal and vertical planes. RESULTS: One-hundred and thirty six professionals participated in the study, and only 25.4% (95% CI 18.5-32.9) of them placed both electrodes within 5 cm from the recommended position. The majority of the participants placed the apical electrode too anteriorly. Of the participants, 36.0% were not aware of the new guidelines. Awareness of the guidelines did not increase the accuracy in electrode placement. CONCLUSIONS: The publication of the national evidence based resuscitation guidelines did not seem to have influenced the practice of placement of the defibrillation electrodes to any major extent. The correct placement of the electrodes needs be emphasized more in the resuscitation training. PMID- 15135191 TI - Airway management in cardiac arrest--comparison of the laryngeal tube, tracheal intubation and bag-valve mask ventilation in emergency medical training. AB - Tracheal intubation (ETI) is considered the method of choice for securing the airway and for providing effective ventilation during cardiac arrest. However, ETI requires skills which are difficult to maintain especially if practised infrequently. The laryngeal tube (LT) has been successfully tested and used in anaesthesia and in simulated cardiac arrest in manikins. To compare the initiation and success of ventilation with the LT, ETI and bag-valve mask (BVM) in a cardiac arrest scenario, 60 fire-fighter emergency medical technician (EMT) students formed teams of two rescuers at random and were allocated to use these devices. We found that the teams using the LT were able to initiate ventilation more rapidly than those performing ETI (P < 0.0001). The LT and ETI provided equal minute volumes of ventilation, which was significantly higher than that delivered with the BVM (P < 0.0001). Our data suggest that the LT may enable airway control more rapidly and as effectively as ETI, and compared to BVM, may provide better minute ventilation when used by inexperienced personnel. PMID- 15135193 TI - The attitude of cardiac care patients towards CPR and CPR education. AB - The recommended targeting of the elderly, those with heart conditions and their family members for CPR education remains unaccomplished. Little is known about cardiac patients' knowledge of and attitude towards CPR and CPR education. This study aimed to investigate cardiac care patients' attitude towards CPR and interest in CPR education. An interview, based on a questionnaire, was conducted with 401 consecutive patients admitted to a coronary care unit. Most participants had heard about the concept of CPR and 64% were aware of its content. In the event of an emergency, 96% were willing to undergo CPR. Age, previous myocardial infarction and heart failure were significantly associated with the willingness or lack of willingness to undergo CPR. Forty percent of the participants had attended one or more courses but only a few within the last two years. The major reasons for not being educated in CPR were a lack of awareness of the availability of CPR training for the public, lack of interest or lack of enterprise. Among those not educated in CPR, 46% would like to attend a course. A hospital was the preferred location for the course, often due to the perceived higher competence of the instructors, but sometimes, because it offered a safe environment. The primary health care centre was preferred because of its location near the participants' homes. In order to increase the proportion of people trained in CPR in target groups such as cardiac care patients and their family members, healthcare professionals should provide patients with information and opportunities to attend locally situated, professionally led courses. PMID- 15135192 TI - A comparison between over-the-head and standard cardiopulmonary resuscitation. AB - BACKGROUND: Standard cardiopulmonary resuscitation (CPR) is performed by kneeling adjacent to the side of the casualty. In certain circumstances it may be difficult or impossible to perform CPR in this position, for example in confined spaces such as a narrow corridor, aircraft or train gangway. The aim of this study was to investigate the technique of over-the-head CPR (OTH CPR), where the CPR provider kneels above the casualty and performs chest compressions OTH of the casualty. METHODS: Twenty volunteers were randomised to a cross over trial where they performed standard and OTH CPR at a 7-day interval. Compression and ventilation variables were recorded on the Laerdal Resusci Annie VAM system. RESULTS: Chest compression depth and ventilation volume declined over time (0-3 min P < 0.001). There was no difference in compression rate, depth, duty cycle or ventilation rate, inflation rate and ventilation volume between techniques. Hand position was incorrect more frequently in the standard compared to the OTH group (incorrect compressions 300 versus 76, respectively, P < 0.001) due principally to a greater proportion of low positioned compressions in the standard CPR group. CONCLUSION: OTH CPR appears equally effective as standard CPR with some marginal advantages in correct hand placement. We suggest that in situations where it is not possible to perform standard CPR, OTH CPR may be considered as a suitable alternative. PMID- 15135194 TI - Optimizing chest compression to rescue ventilation ratios during one-rescuer CPR by professionals and lay persons: children are not just little adults. AB - OBJECTIVE: To estimate the optimum ratio of chest compressions to ventilations for one-rescuer CPR that maximizes systemic oxygen delivery in children. METHOD: Equations describing oxygen delivery and blood flow during CPR as functions of the number of compressions and the number of ventilations delivered over time were adapted from the former work of Babbs and Kern. These equations were solved explicitly as a function of body weight, using scaling algorithms based upon principles of developmental anatomy and physiology. RESULTS: The optimal compression to ventilation (C/V) ratios for infants and younger children increase sharply as a function of body weight. Optimal C/V ratios are lower for professional rescuers, who take less time to deliver a rescue breath, than for lay rescuers, who interrupt chest compressions for longer to perform ventilations. For professional rescuers the optimal C/V ratio, x*, is approximately 1.6 square root W where the W is the patient's body weight in kg. For lay rescuers the optimum C/V ratio is approximately 2.8 square root W. These values can be approximated for children and teens by the following rules of thumb, based upon the age of the victim: "5 + one half the age in years" for professional rescuers and "5 + age in years" for lay rescuers. CONCLUSIONS: Compression to ventilation ratios in CPR should be smaller for children than for adults and gradually increase as a function of body weight. Optimal CPR in children requires relatively more ventilation than optimal CPR in adults. A universal compression/ventilation ratio of 50:2, targeted to optimize adult resuscitation, would not be appropriate for infants and young children. PMID- 15135195 TI - Role of semiautomatic defibrillators in a general hospital: "Naples Heart Project". AB - In Italian hospitals, 85% of patients hospitalized in general medical wards who experience cardiac arrest die, while the incidence is much lower in patients in intensive care units. Defibrillation, in Italian hospitals, often occurs very late, either due to a lack of defibrillators, or due to architectural and structural barriers. The object of an in-hospital emergency service is to prevent and treat cardiac arrest without subsequent complications, such as brain damage, renal failure etc. The Naples Heart Project was based on a feasibility study of the in-hospital emergency service to evaluate and analyze problems associated with type of structure, departmental and institutional dislocation, internal practicability (architectural features and preferential ways), staff numbers and distribution, the calling system for emergency, and the equipment available. The Naples Heart Project began in July 2001, since then it has already created 835 BLSD first responders among the hospital staff; 440 were physicians and physicians still in training, 310 were nurses and 85 were administrative staff. PMID- 15135196 TI - Attenuated adult biphasic shocks compared with weight-based monophasic shocks in a swine model of prolonged pediatric ventricular fibrillation. AB - AIM: To compare the safety and efficacy of attenuated adult biphasic shocks with standard monophasic weight-based shocks in a piglet model of prolonged prehospital ventricular fibrillation (VF). BACKGROUND: If attenuated adult shocks are safe and effective for prehospital pediatric VF, automated external defibrillators (AEDs) can be easily adapted for pediatric use. METHODS: After 7 min of untreated VF, piglets were randomized to treatment with attenuated adult biphasic shocks or weight-based monophasic shocks. The attenuated adult biphasic group received 200/300/360 J shocks, attenuated by specialized pediatric electrodes to 51/78/81 J and the monophasic weight-based control group received 2/4/4 J/kg shocks. Forty-eight female piglets were studied, 16 in each of three weight categories: 4 kg (neonatal), 14 kg (younger child) and 24 kg (older child). The primary outcome measures of efficacy and safety were 24h survival with good neurological outcome and post-resuscitation left ventricular ejection fraction (LVEF), respectively. RESULTS: For the 24 kg piglets, attenuated adult biphasic shocks resulted in superior 24 h survival with good neurological outcome (6/8 versus 0/8, P < 0.001) and greater LVEF 4 h post-resuscitation (34 +/- 4% versus 18 +/- 5%, P < 0.05). For the 14 and 4 kg piglets, 24 h survival with good neurological outcome occurred in 7/8 versus 5/8 and 7/8 versus 3/8, respectively, and LVEF 4 h post-resuscitation was 30 +/- 3% versus 36 +/- 6% and 30 +/- 3% versus 22 +/- 4%, respectively. CONCLUSIONS: The escalating attenuated adult biphasic dosage strategy was at least as safe and effective as the standard weight-based monophasic dose over a wide range of weights in this piglet model of prehospital VF. This work supports the concept of using an attenuated adult biphasic dosage in children. PMID- 15135197 TI - Optimal dosing of dobutamine for treating post-resuscitation left ventricular dysfunction. AB - OBJECTIVES: This study was designed to determine the optimal dose of dobutamine in the treatment of post-resuscitation left ventricular dysfunction. BACKGROUND: Global left ventricular dysfunction following successful resuscitation from prolonged, ventricular fibrillation cardiac arrest, negatively impacts long-term survival. Dobutamine can overcome this global myocardial stunning. Previous data indicate a dose of 10 mcg/kgmin improves systolic and diastolic function, but markedly increases the heart rate. METHODS: Twenty swine (24 +/- 0.4 kg) were randomized to one of four doses (0, 2, 5, and 7.5 mcg/kgmin) of dobutamine for the treatment of post-resuscitation myocardial dysfunction following 12.5 min of untreated ventricular fibrillation cardiac arrest. Cardiac function was measured at pre-arrest baseline and serially for 6 h post-resuscitation. Left ventricular function was evaluated by contrast ventriculograms, left ventricular pressures, +dP/dt, Tau, -dP/dt, and cardiac output. Myocardial oxygen consumption and myocardial blood flow were measured to assess the functional significance of any dobutamine-mediated heart rate responses. RESULTS: Left ventricular dysfunction was evident at 25 min and peaked 4 h post-resuscitation. Significant (P < 0.05) improvements in ventricular systolic (EF, CO) and diastolic (LVEDP, Tau) function were evident within minutes of dobutamine initiation and persisted at 6h for the 5 and 7.5 mcg/kgmin groups. Tachycardia manifested with all dobutamine doses, but only affected myocardial oxygen consumption significantly (P < 0.05) at the highest dose (7.5 mcg/kgmin). CONCLUSIONS: Dobutamine at 5 mcg/kgmin appears optimal for restoring systolic and diastolic function post-resuscitation without adversely affecting myocardial oxygen consumption. PMID- 15135198 TI - Effect of ifenprodil, a polyamine site NMDA receptor antagonist, on brain edema formation following asphyxial cardiac arrest in rats. AB - OBJECTIVE: Brain edema occurs in experimental and clinical cardiac arrest (CA) and is predictive of a poor neurological outcome. N-Methyl--aspartate (NMDA) receptors contribute to brain edema elicited by focal cerebral ischemia/reperfusion (I/R). Ifenprodil, a NMDA receptor antagonist, attenuates brain edema and injury size in rats after focal cerebral I/R. We assessed the hypothesis that ifenprodil reduces CA-elicited brain edema. METHODS: Eighteen male Sprague-Dawley rats were assigned to group 1 (normal control, n=6), group 2 (placebo-treated CA, n=6), or group 3 (ifenprodil-treated CA, n=6). CA was induced by 8 min of asphyxiation and the animals were resuscitated with cardiopulmonary resuscitation (CPR), ventilation, epinephrine (adrenaline), and sodium bicarbonate (NaHCO3). Ifenprodil of 10 mg/kg or a placebo vehicle was given intraperitoneally 5 min before CA. Brain edema was determined by brain wet to-dry weight ratio at 1 h after resuscitation. RESULTS: There were no differences between groups 2 and 3 in all physiological variables at baseline. Time from asphyxiation to CA was 201.5 +/- 7.5 s in group 2 and 160.7 +/- 10.4 s in group 3 (P<0.001). Resuscitation time was 68.2 +/- 13.3 s in group 2 and 92.8 +/- 18.2 s in group 3 (P<0.05). Ifenprodil decreased mean arterial pressure (MAP) before asphyxiation, from 128 +/- 7 in group 2 to 82 +/- 15 mmHg in group 3 (P<0.001), and negated immediate post-resuscitation hypertension. Brain wet-to dry weight ratio was 5.64 +/- 0.44 in group 1, 7.34 +/- 0.95 in group 2 (P<0.01 versus group 1), and 5.93 +/- 0.40 in group 3 (P<0.05 versus group 2). CONCLUSIONS: Ifenprodil reduces CA-elicited brain edema. In addition, we observed significant hemodynamic changes caused by ifenprodil. PMID- 15135199 TI - Influence of selective nitric oxide synthetase inhibitor for treatment of refractory haemorrhagic shock. AB - OBJECTIVE: Haemorrhagic shock (HS) is implicated in the induction of inducible nitric oxide synthase that leads to increased production of nitric oxide (NO). We investigated the influence of aminoguanidine (AG), a selective iNOS inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME), a non-selective inhibitor and S Nitroso-N-acetylpenicillamine (SNAP), a NO donor, each of which was given with (+) or without (-) angiotensin II (ANGII), a vasoconstrictor, on the survival rate of HS decompensatory phased (HSDP) rats. MATERIALS AND METHODS: HSDP was achieved via a constant pressure method. Organs were harvested and analyzed from rats sacrificed 72 h after HSDP or upon death. Plasma collected from HSDP rats were used to measure nitrate/nitrite, GOT and creatinine levels. RESULTS: AG+ANGII-treated rats had significantly higher survival rates compared to the other treatment groups, 72 h following HSDP. A marked increase in MABP level was observed in AG+ANGII-treated rats when compared to other treatment groups. Histological examinations also showed a reduction of organ damage in AG+ANGII treated rats compared to other treatment groups. Nitrate/nitrite level, glutamic oxalacetic transaminase (GOT) level and creatinine level were also significantly improved in AG+ANGII-treated rats compared to the other groups. CONCLUSIONS: A greater beneficial effect was achieved with treatment by the AG+ANGII combination. Our experiments showed that the inhibition of excessive NO formation that occurred during HSDP, had augmented the vascular responsiveness effect of ANGII following protracted HS. PMID- 15135200 TI - Rescue percutaneous coronary intervention during cardiopulmonary resuscitation. AB - If sudden cardiac arrest occurs during cardiac catheterization, the underlying coronary condition may be defined immediately by coronary angiography. This may, in turn, allow a lifesaving attempt of percutaneous coronary intervention (PCI). We report on two patients with critical proximal disease of the left coronary artery in whom successful PCI during cardiopulmonary resuscitation (CPR) led to the restoration of a spontaneous circulation and long-term survival. PMID- 15135201 TI - Diving emergencies. PMID- 15135203 TI - Diving emergencies. PMID- 15135204 TI - A wide QRS complex tachycardia following intravenous adenosine. PMID- 15135205 TI - Long QT interval in severe hypercalcaemia. PMID- 15135206 TI - Quantitative relationship between the local lymph node assay and human skin sensitization assays. AB - The local lymph node assay (LLNA) is a new test method which allows for the quantitative assessment of sensitizing potency in the mouse. Here, we investigate the quantitative correlation between results from the LLNA and two human sensitization tests--specifically, human repeat insult patch tests (HRIPTs) and human maximization tests (HMTs). Data for 57 substances were evaluated, of which 46 showed skin sensitizing properties in human tests, whereas 11 yielded negative results in humans. For better comparability data from mouse and human tests were transformed to applied doses per skin area, which ranged over four orders of magnitude for the substances considered. Regression analysis for the 46 human sensitizing substances revealed a significant positive correlation between the LLNA and human tests. The correlation was better between LLNA and HRIPT data (n=23; r=0.77) than between LLNA and HMT data (n=38; r=0.65). The observed scattering of data points is related to various uncertainties, in part associated with insufficiencies of data from older HMT studies. Predominantly negative results in the LLNA for another 11 substances which showed no skin sensitizing activity in human maximization tests further corroborate the correspondence between LLNA and human tests. Based on this analysis, the LLNA can be considered a reliable basis for relative potency assessments for skin sensitizers. Proposals are made for the regulatory exploitation of the LLNA: four potency groups can be established, and assignment of substances to these groups according to the outcome of the LLNA can be used to characterize skin sensitizing potency in substance-specific assessments. Moreover, based on these potency groups, a more adequate consideration of sensitizing substances in preparations becomes possible. It is proposed to replace the current single concentration limit for skin sensitizers in preparations, which leads to an all or nothing classification of a preparation as sensitizing to skin ("R43") in the European Union, by differentiated concentration limits derived from the limits for the four potency groups. PMID- 15135207 TI - Toxicological studies on Lactose Oxidase from Microdochium nivale expressed in Fusarium venenatum. AB - A new carbohydrate oxidase, Lactose Oxidase, with high specificity of oxidizing the disaccharide lactose to lactobionic acid has been found. This enzyme opens up for a variety of applications. A programme of toxicological studies was conducted to establish the safety of Lactose Oxidase to be used as a processing aid in the food industry. The enzyme used in this study was produced by a submerged fermentation of Fusarium venenatum and contained a gene code from Microdochium nivale. Oral administration to rats of up to 10 mL/kg bodyweight (bw)/day (equivalent to a total organic solids dosage of 900 mg/kg bw/day or a Lactose Oxidase dosage of 344 LOXU/kg bw/day) for 13 weeks did not cause any adverse effect. Lactose Oxidase was not found to be mutagenic in the bacterial reverse mutation assay, nor did it cause chromosomal aberrations in cultured human lymphocytes. The maximum recommended dosage of Lactose Oxidase is 50 LOXU/kg liquid whey protein concentrate. The safety margin for exposure is estimated to be at least 6.2 x 10(4) for daily diary product consumption. In conclusion Lactose Oxidase can be considered as safe for use in the food industry. PMID- 15135208 TI - In vitro predictions of skin absorption of caffeine, testosterone, and benzoic acid: a multi-centre comparison study. AB - To obtain better insight into the robustness of in vitro percutaneous absorption methodology, the intra- and inter-laboratory variation in this type of study was investigated in 10 European laboratories. To this purpose, the in vitro absorption of three compounds through human skin (9 laboratories) and rat skin (1 laboratory) was determined. The test materials were benzoic acid, caffeine, and testosterone, representing a range of different physico-chemical properties. All laboratories performed their studies according to a detailed protocol in which all experimental details were described and each laboratory performed at least three independent experiments for each test chemical. All laboratories assigned the absorption of benzoic acid through human skin, the highest ranking of the three compounds (overall mean flux of 16.54+/-11.87 microg/cm(2)/h). The absorption of caffeine and testosterone through human skin was similar, having overall mean maximum absorption rates of 2.24+/-1.43 microg/cm(2)/h and 1.63+/ 1.94 microg/cm(2)/h, respectively. In 7 out of 9 laboratories, the maximum absorption rates of caffeine were ranked higher than testosterone. No differences were observed between the mean absorption through human skin and the one rat study for benzoic acid and testosterone. For caffeine the maximum absorption rate and the total penetration through rat skin were clearly higher than the mean value for human skin. When evaluating all data, it appeared that no consistent relation existed between the diffusion cell type and the absorption of the test compounds. Skin thickness only slightly influenced the absorption of benzoic acid and caffeine. In contrast, the maximum absorption rate of testosterone was clearly higher in the laboratories using thin, dermatomed skin membranes. Testosterone is the most lipophilic compound and showed also a higher presence in the skin membrane after 24 h than the two other compounds. The results of this study indicate that the in vitro methodology for assessing skin absorption is relatively robust. A major effort was made to standardize the study performance, but, unlike in a formal validation study, not all variables were controlled. The variation observed may be largely attributed to human variability in dermal absorption and the skin source. For the most lipophilic compound, testosterone, skin thickness proved to be a critical variable. PMID- 15135209 TI - Role of biokinetics in risk assessment of drugs and chemicals in children. AB - Whether children incur different risks from xenobiotics than adults will depend on the exposure, biokinetics, and dynamics of compound. In this paper, current knowledge on developmental physiology and possible effects on biokinetics are evaluated and the role of biokinetics in risk assessment both for drugs and chemicals is discussed. It is concluded that most dramatic age-related physiological changes that may affect biokinetics occur in the first 6-12 months of age. The difference in internal exposure between children and adults can generally be predicted from already known developmental physiological differences. However, for risk assessment it will also be necessary to determine whether internal exposure is within the drug's therapeutic window or if it will exceed the NOAEL of a chemical. Furthermore, the effects of internal exposure of potentially harmful compounds on developing organ systems is of utmost importance. However, knowledge on this aspect is very limited. Risk assessment in children could be improved by: (1) application of pediatric PBPK-models in order to gain insight into internal exposure in children, (2) studies in juvenile animals for studying effects on developing systems, and (3) extrapolation of knowledge on the relationship between internal exposure and dynamics for drugs to other chemicals. PMID- 15135210 TI - Mammalian toxicology overview and human risk assessment for sulfosulfuron. AB - Sulfosulfuron is a low-use rate sulfonylurea herbicide. A review of the toxicity database for sulfosulfuron indicates that the molecule has a low order of acute toxicity. It is not genotoxic and is not a reproductive, developmental, or nervous system toxicant. There were no indications of endocrine disruption in any study performed with the molecule. The only findings considered to be an adverse effect in mammalian laboratory animals following prolonged subchronic or chronic exposure to sulfosulfuron were isolated to the urinary tract. These findings occurred in conjunction with findings of urolith formation following high-level chemical dosing, resulting in epithelial hyperplasia that, in a few cases, progressed to tumor formation. Mode-of-action information supports the conclusion that these tumors result from a non-genotoxic, threshold-based process that is well established and widely considered to be not relevant to humans. Based on its short-term, infrequent application pattern and very low use rate and crop residues, aggregate and cumulative risk assessments indicate that sulfosulfuron has substantial margins of exposure and does not represent a significant risk to human health. PMID- 15135211 TI - Effect of strain and diet upon constitutive and chemically induced activities of several xenobiotic-metabolizing enzymes in rats. AB - The response of animals in toxicity studies reflects a complex interaction of a number of variables, some intrinsic to a particular study design and others resulting from the treatment itself. The influences of strain and diet upon constitutive and benzo(a)pyrene (B(a)P) induced activities of several hepatic Phase I and II enzymes were studied in a multifactoral design. Male and female CDF and Crl:CD rats were fed a standard rodent diet ad libitum, a 75% of ad libitum restricted feeding regimen or a phytoestrogen-free diet for approximately 3 weeks. During the last five days of the study, rats were administered either corn oil (vehicle) or 15 mg/kg/day B(a)P via oral gavage. The constitutive activities of hepatic CYP1A1, CYP1A2, CYP2B1/2, and mixed isoforms of UDP glucuronosyl transferase, sulfotransferase, and glutathione-S-transferase varied significantly by feeding regimen and strain. Responses to B(a)P administration were also observed to be influenced by diet and strain in a manner similar to that observed for constitutive activities. These findings point out the potentially significant interactions of relatively commonly encountered variables that may affect results of hazard testing, especially when employing near metabolically saturating dosages of test chemicals. PMID- 15135212 TI - Allometric principles for interspecies extrapolation in toxicological risk assessment--empirical investigations. AB - Four types of data (toxicokinetic data of pharmaceuticals from six species including humans, LD(50) values from eight animal species, long-term NOAEL values of pesticides from mice, rats, and dogs, and toxicity data on anti-neoplastic agents from six species including humans) were used for interspecies comparisons. Species differences with regard to kinetic parameters and toxicity were evaluated and the concordance with predictions by allometric scaling according to caloric demand (allometric exponent 0.75) or to body weight (allometric exponent 1) was checked. For LD(50) values, agreement was poor for both allometric concepts. Recently reported concordance of LD(50) species differences with body weight scaling could be traced back to biased data selection. The other three datasets are clearly in agreement with the allometric scaling according to caloric demand. Caloric demand scaling is thus proposed as a generic interspecies extrapolation method in the absence of substance-specific data. Moreover, the evaluated data make it possible to describe uncertainty associated with the process of interspecies extrapolation by allometric rules. PMID- 15135213 TI - Interspecies differences in susceptibility to perturbation of thyroid homeostasis: a case study with perchlorate. AB - Despite many physiological similarities, humans and rats exhibit notably different susceptibilities to thyroid perturbation. Considerable research has recently been conducted on the thyroid-active chemical perchlorate, a chemical of emerging environmental and regulatory interest. While the data indicate humans and rats exhibit similar dose-response relationships in terms of acute inhibition of thyroidal iodide uptake, the two species appear to exhibit notable differences in terms of thyroid hormone response, the toxicologically significant consequence of iodide uptake inhibition. We analyzed dose-response data for changes in serum T(3), T(4), and TSH levels from studies in humans, rats, mice, and rabbits. We found that thyroid homeostasis in the rat appears to be strikingly more sensitive to perchlorate than any of the other species. Rats exhibited an increase in serum TSH at 0.1mg/kg-day whereas other species remained unresponsive even at doses of 10mg/kg-day. Less pronounced but consistent effects were seen with serum T(3) and T(4). These cross-species comparisons provide strong evidence that data obtained from rat studies should be critically evaluated for their relevance to humans. If rat data are used to develop toxicity criteria for perchlorate, we propose that this is an instance where an inter-species uncertainty factor less than one is supportable. DISCLOSURE STATEMENT: One of the authors (BDB) has been hired by Lockheed Martin Corporation as an expert in litigation involving perchlorate. A portion of the initial research presented in this paper was conducted in conjunction with her role in that matter. PMID- 15135215 TI - Developing drugs for pediatric use: a role for juvenile animal studies? AB - The increased interest and/or need to perform pediatric clinical trials to allow the marketing and safe use of a wider range of medicines in children has raised the profile of the need to conduct juvenile animal studies. It is argued that such studies may identify "unique" toxicities not seen from available adult animal and clinical data. This paper will review the current situation from an industrial, regulatory, and scientific perspective. Areas of important consideration include functional (dynamic) and kinetic differences between children and adults and specific considerations associated with testing in young animals. This paper will also review what are we currently doing? Whether we really need these studies? and What challenges lie in the future? PMID- 15135214 TI - Characterization of risk for general population exposure to perfluorooctanoate. AB - Perfluorooctanoate (PFOA), an environmentally and metabolically stable perfluorinated carboxylic acid, has been detected in the serum of children, adults and the elderly from the United States with the upper bound of the 95th percentile estimate in the range of 0.011-0.014 microg/mL (ppm). In this risk characterization, margins of exposure (MOE), which can provide a realistic perspective on potential for human risk, were determined by comparison of general population serum PFOA concentrations with serum concentrations from toxicological studies that are associated with the lower 95% confidence limit of a modeled 10 percent response or incidence level (LBMIC(10)) using USEPA BMDS software. The LBMIC(10) was estimated using surrogate data from other studies or pharmacokinetic relationships if serum PFOA data were not available. Modeled dose responses (with resulting LBMIC(10) values) included post-natal effects in rats (29 microg/mL), liver-weight increase (23 microg/mL), and body-weight change (60 microg/mL) in rats and monkeys, and incidence of Leydig cell adenoma (125 microg/mL) in rats. MOE values based on the upper bound 95th percentile population serum PFOA concentration were large, ranging from 1600 (liver-weight increase) to 8900 (Leydig cell adenoma). These MOE values represent substantial protection of children, adults, and the elderly. PMID- 15135216 TI - Safety evaluation of a natural tomato oleoresin extract derived from food processing tomatoes. AB - Experimental and epidemiological studies indicate that consumption of tomato products containing high amounts of lycopene is associated with lowered cancer risk. The protective effects of lycopene may be related to its antioxidant potential. Lycopene has been demonstrated to inhibit oxidation. A proprietary, natural tomato oleoresin extract (NTOE), is a purified tomato oleoresin containing 6% lycopene produced from tomatoes. NTOE was evaluated for toxicological effects, and found the 50% lethal dose (LD(50)), derived from the acute oral toxicity study, was greater than 5000 mg/kg body weight. The no observed-adverse-effect level (NOAEL) derived from the 13-week study was 4500 mg/kg/day. Acute dermal toxicity study of NTOE found no toxicity at 2000 mg/kg body weight. NTOE lacked dermal irritation in the rabbit model, but was found to have moderate eye-irritant capabilities. NTOE tested at 5% (w/w) in petroleum jelly was a moderate sensitizer in the guinea pig model. There was no evidence of mutagenic potential up to 5000 microg/plate, as determined by the Ames assay. These results demonstrate the inability of NTOE to produce oral, dermal or mutagenic toxicity in animal models at doses greater than 300 times the normal human consumption of lycopene. Consumption analysis of lycopene-containing foods estimated mean daily intake of lycopene at 8.2mg/day. PMID- 15135217 TI - Synergistic effect of dexamethasone and beta-adrenergic receptor agonists on the nerve growth factor gene transcription. AB - Activation of beta-adrenergic receptor (betaAR) increases the synthesis of nerve growth factor (NGF) in the brain and in C6-2B glioma cells. However, in the brain, the betaAR-mediated increase in NGF expression appears to require the presence of glucocorticoids, suggesting that NGF promoter may be sensitive to cAMP and glucocorticoid-dependent transcription factors. We tested this hypothesis by exposing C6-2B glioma cells to dexamethasone (DEX) in combination with agents that increase cAMP levels and examining the DNA binding activity of two cAMP-dependent transcription factors that regulate NGF expression: cAMP responsive element binding protein (CREB) and CCAAT/enhancer binding protein delta (C/EBPdelta). Electrophoretic mobility shift assays revealed that the beta(2)AR agonist clenbuterol (CLE) or high levels of cAMP elicited a time dependent increase in C/EBPdelta binding activity as well as phosphorylated CREB (P-CREB). When DEX, which per se showed little effect on these transcription factors, was combined with CLE, dibutyryl cAMP or isoproterenol, enhanced induction of P-CREB and C/EBP binding activity as well as NGF mRNA was observed. Moreover, the increase in NGF mRNA in the presence of DEX was prolonged compared to that obtained by CLE or other cAMP inducing agents alone. In fact, NGF mRNA levels remained significantly elevated at least for 24 h. These studies suggest that the synergistic effect of DEX on the induction of NGF mRNA may include the ability of this glucocorticoid to potentiate the betaAR-mediated induction of transcription factors. PMID- 15135218 TI - The identification of a second actin-binding region in spinophilin/neurabin II. AB - Spinophilin/neurabin II is an actin-associated scaffolding protein enriched in the dendritic spines of neurons. Previously, the actin-binding domain (ABD) of spinophilin was localized to a domain between amino acids (aa) 1 and 154. In a mass spectrometry screen for spinophilin-binding proteins, we have identified an additional actin-binding region between aa 151 and 282. F-actin co-sedimentation and GST affinity chromotography experiments further substantiate this result. Phalloidin staining of Rat2 fibroblasts transiently expressing GFP-spinophilin deletion constructs indicates co-localization with a subset of actin. Regions of spinophilin that lack the revised ABD (aa 1-230) do not co-localize with phalloidin-labeled actin, suggesting that the actin-binding domain contributes to directing the subcellular distribution of spinophilin. Targeting experiments using primary hippocampal cultures indicate that only the first actin-binding site contributes to dendritic spine localization. The second ABD targets to spines inefficiently and thus may interact with and affect actin filaments in a different manner than the first ABD. PMID- 15135219 TI - Loss of glial fibrillary acidic protein results in decreased glutamate transport and inhibition of PKA-induced EAAT2 cell surface trafficking. AB - Loss of the astrocyte-specific intermediate filament protein, glial fibrillary acidic protein (GFAP) results in an increased susceptibility to ischemic insult, enhanced hippocampal LTP, and decreased cerebellar long-term depression (LTD). Because glutamate receptor activation plays a key role in cell death and cellular plasticity responses, we wanted to determine if alterations in glial glutamate transport could contribute to the GFAP null phenotype. To address functional changes in glutamate transport, we measured glutamate uptake in cortical, cerebellar, and hippocampal synaptosomal preparations from age-matched adult wild type and GFAP null mice and demonstrated a 25-30% reduction in the V(max) for d aspartate uptake in the cortex and hippocampus of GFAP null animals. Western blot analysis of cortical synaptosomal fractions from wild type and GFAP null animals demonstrated that loss of GFAP results in decreases in both astrocytic (EAAT1) and neuronal (EAAT3) glutamate transporter subtypes. Immunohistochemical analysis demonstrated a region-specific modification of neuronal glutamate transporter, EAAT3 trafficking in the GFAP null phenotype. Analysis of primary cortical astrocyte cultures prepared from GFAP null and wild type mice demonstrated that loss of GFAP results in an inability to traffic the glial glutamate transporter, EAAT2, to the surface of the cell following protein kinase A (PKA) stimulation by dibutyryl cAMP. Taken together, these results suggest that the intermediate filament protein, GFAP plays a key role in modulating astrocytic and neuronal glutamate transporter trafficking and function. PMID- 15135220 TI - Molecular cloning and partial functional characterization of Tsha3--a novel modulatory potassium channel alpha-subunit of trout CNS. AB - A novel Shaker-related potassium channel subunit termed Tsha3 that is widely expressed in the CNS of trout was PCR-cloned and sequenced: its deduced amino acid sequence showed an extended N-terminal domain with a high proportion of negatively charged residues and possessed highest similarity with KCNA10, a human epithelial potassium channel. Upon heterologous expression in Sf21 cells, homomeric Tsha3 did not yield voltage-activated potassium channels but produced only ohmic currents that reversed at -15 mV. After co-expression with Tsha1, a novel outward rectifier current was generated that differed from homomeric Tsha1 by its slower kinetics of activation, its partial current inactivation, and its partial blockade by 5 mM TEA as well as 1 microM DTX. Co-immunoprecipitation studies using anti-Tsha3 antibodies confirmed that Tsha3 tightly bound with Tsha1 in co-infected Sf21 cells. As revealed from GFP- and DsRed-labeling studies, the pattern of distribution of Tsha1 was profoundly altered after co-infection with Tsha3 subunits. PMID- 15135221 TI - Opiate-like effects of sugar on gene expression in reward areas of the rat brain. AB - Drugs abused by humans are thought to activate areas in the ventral striatum of the brain that engage the organism in important adaptive behaviors, such as eating. In support of this, we report here that striatal regions of sugar dependent rats show alterations in dopamine and opioid mRNA levels similar to morphine-dependent rats. Specifically, after a chronic schedule of intermittent bingeing on a sucrose solution, mRNA levels for the D2 dopamine receptor, and the preproenkephalin and preprotachykinin genes were decreased in dopamine-receptive regions of the forebrain, while D3 dopamine receptor mRNA was increased. While morphine affects gene expression across the entire dopamine-receptive striatum, significant differences were detected in the effects of sugar on the nucleus accumbens and adjacent caudate-putamen. The effects of sugar on mRNA levels were of greater magnitude in the nucleus accumbens than in the caudate-putamen. These areas also showed clear differences in the interactions among the genes, especially between D3R and the other genes. This was revealed by a novel multivariate analysis method that identified cooperative interactions among genes, specifically in the nucleus accumbens but not the caudate-putamen. Finally, a role for these cooperative interactions in a load-sharing response to perturbations caused by sugar was supported by the finding of a different pattern of correlations between the genes in the two striatal regions. These findings support a major role for the nucleus accumbens in mediating the effects of naturally rewarding substances and extend an animal model for studying the common substrates of drug addiction and eating disorders. PMID- 15135222 TI - Daily and circadian expression of neuropeptides in the suprachiasmatic nuclei of nocturnal and diurnal rodents. AB - The suprachiasmatic nuclei (SCN) of the hypothalamus are necessary for coordination of major aspects of circadian rhythmicity in mammals. Although the molecular clock mechanism of the SCN has been a field of intense research during the last decade, the role of the neuropeptides in the SCN, including arginine vasopressin (AVP), vasoactive intestinal polypeptide (VIP) and gastrin-releasing peptide (GRP), in the clock itself or in circadian organization is still largely unknown. Previous studies mainly performed in the rat have examined the profiles of AVP, VIP and GRP mRNA and peptide levels and suggested that the AVP rhythm is controlled by the circadian clock, whereas those of VIP and GRP are directly dependent on lighting conditions. Here, both daily (i.e., under light-dark cycle [LD]) and circadian (i.e., in constant darkness [DD]) profiles of neuropeptide mRNA were investigated in the SCN of the nocturnal mouse Mus musculus and the diurnal rodent Arvicanthis ansorgei to gain insight into a possible role in circadian organization. Our data show that AVP mRNA exhibits a clear circadian rhythm in the SCN peaking by the end of the subjective day in both species. Contrary to what has been observed in rats, oscillations of VIP and GRP mRNA in the SCN are found to be clock-controlled in mice and A. ansorgei, but with different phases for peak expression. While both VIP and GRP mRNA peak during the middle of the subjective night (i.e., with a 6-h lag compared to AVP mRNA) in mice, they peak almost in phase with AVP mRNA in A. ansorgei. Contrary to what has been reported in the rat, mean levels of VIP and GRP peptide mRNA levels tended to be increased by light in the mice. The different circadian organization of SCN neuropeptides mRNA profiles in both light/dark and constant darkness conditions between mice and A. ansorgei could be related with diurnality. PMID- 15135223 TI - Environmental light conditions alter gene expression of rat catecholamine biosynthetic enzymes and Neuropeptide Y: differential effect in superior cervical ganglia and adrenal gland. AB - The hypothalamic suprachiasmatic nuclei (SCN) comprise the main site in the brain involved in the control of the homeostatic mechanism which respond to environmental daily light changes. The sympathetic nervous system and hypothalamic releasing or inhibiting factors mediate the SCN control of a number of peripheral organs and tissues. In this work we analyzed the involvement of two environmental light conditions, constant light (LL) and constant dark (DD) for 20 days, on the expression of mRNAs for catecholamines biosynthetic enzymes and neuropeptide Y (NPY) genes in rat superior cervical ganglia (SCG) and adrenal gland. The results of Northern blot analysis show that LL exposure reduces mRNA levels for tyrosine hydroxylase (TH) the rate limiting catecholamine biosynthetic enzyme and also of dopamine beta-hydroxylase (DBH) as well as for NPY in SCG to about half the levels in control animals. In contrast, exposure of the rats to DD did not elicit any change in the SCG. In the adrenal gland, both, LL and DD conditions increased the TH, DBH as well as phenylethanolamine N methyltransferase (PNMT) mRNA levels. Under the same conditions, adrenal NPY mRNA levels were decreased by either LL or DD. The results show, for the first time, that prolonged changes in environmental light can alter the gene expression of catecholamine biosynthetic enzymes and of NPY. There was differential response in SCG and adrenal gland. PMID- 15135224 TI - Bacterial DNA induced iNOS expression through MyD88-p38 MAP kinase in mouse primary cultured glial cells. AB - To study the role of bacterial DNA in the immune function of the brain, we examined the effect of CpG-DNA on the inducible nitric oxide synthase (iNOS) expression in mouse primary cultured glial cells. The expression of Toll-like receptor 9 (TLR9), the receptor of bacterial DNA, was detected by RT-PCR. We observed an increase in iNOS mRNA 6 h after CpG-DNA application. The expression of iNOS protein peaked at 12 h and declined thereafter. CpG-DNA increased p38 mitogen-activated protein kinase (MAPK) activation in primary cultured glial cells. SB203580, a specific inhibitor of p38 MAP kinase, inhibited the CpG-DNA induced iNOS expression. Moreover, CpG-DNA failed to activate p38 MAP kinase and iNOS induction in the primary cultured glial cells prepared from myeloid differentiation factor 88 (MyD88) deficient mice. Therefore, it is suggested that functional receptor for bacterial DNA exists in primary cultured glial cells and CpG-DNA induces iNOS expression via the MyD88-p38 MAP kinase-dependent mechanisms. Thus, the present results point to the important role of bacterial DNA by acting on glial cells to operate brain immune function. PMID- 15135225 TI - Regulation of pontine neurite morphology by target-derived signals. AB - The molecular cues that regulate neurite morphology within the target environment are key to the formation of complex neural circuitry. During development of the ponto-cerebellar projection, pontine fibers sprout and form elaborate arbors within the inner cerebellar layer prior to arrival of their target cells, the cerebellar granule neurons. Here, we describe the biochemical fractionation of two granule neuron-derived factors that stimulate elaboration of pontine neurites. These factors were identified using a dissociated pontine bioassay and biochemically fractionated from granule cell (GC) conditioned medium (GCCM). One of the factors, STIM1, is a protein with a molecular weight greater than 30 kDa that is distinct from known neurotrophins. The other, STIM2, is a small, protease resistant molecule with an estimated molecular weight below 1 kDa. We show that these factors stimulate pontine neurite elongation both independently and cooperatively and thus may contribute to the formation of elaborate pontine arbors within the cerebellar cortex. PMID- 15135226 TI - Circadian rhythms in behavior and clock gene expressions in the brain of mice lacking histidine decarboxylase. AB - To clarify functional roles of histamine in the circadian clock system, circadian rhythms of behavior and clock gene expression in the brain were examined in the mouse lacking histidine decarboxylase (HDC-/- mouse). Wheel-running and spontaneous locomotion were recorded under light-dark cycle (LD) and constant darkness (DD). mPer1, mPer2 and mBMAL1 mRNA expression rhythms under LD and DD were measured in the suprachiasmatic nucleus (SCN), cerebral cortex and striatum by in situ hybridization. The activity levels under LD and DD in the HDC-/- mice were lower than that in the wild type regardless of activity types (wheel-running and spontaneous locomotion). The free-running period under DD was significantly longer in the HDC-/- mice than in the wild type. The 24-h profiles of mPer1, mPer2 and mBMAL1 mRNA expressions in the SCN were not different between the two genotypes. By contrast, the mPer1 and mPer2 mRNA rhythms in the other brain areas such as the cortex and striatum were significantly disrupted in the HDC-/- mice. These results suggest that histamine is involved in the circadian system especially in the output pathway or feedback route from behavior to the pacemaker in the SCN, and that mPer genes in the brain areas outside the SCN play an important role in the expression of behavioral rhythm. PMID- 15135227 TI - Nicotine attenuates oxidative stress, activation of redox-regulated transcription factors and induction of proinflammatory genes in compressive spinal cord trauma. AB - Pathophysiology of neurodegeneration following spinal cord injury (SCI) involves alterations of cellular redox status, activation of transcription factors and induction of proinflammatory genes. In addition, recent evidence indicates that nicotine can induce potent neuroprotective effects. To study the influence of nicotine on the redox signaling pathways in relationship to SCI, moderate contusions of spinal cords at the level of T-10 were induced in rats treated or untreated with nicotine. Cellular oxidative stress, DNA binding activity of redox responsive transcription factors (AP-1, NF-kappaB and CREB) as well as mRNA levels of inflammatory genes (MCP-1 and TNF-alpha) were determined in the thoracic and lumbar regions of the spinal cords. Nicotine was administrated 2 h after the SCI in a single i.p. injection at the dose of 0.35, 3.5 or 7 mg/kg, and rats were sacrificed 3 h following such an injection. Spinal cord trauma was associated with a significant increase in oxidative stress, and activation of NF kappaB, AP-1 and CREB, as well as overexpression of MCP-1 and TNF-alpha in both the thoracic and lumbar regions. Nicotine administration following the SCI markedly attenuated, especially in the lumbar region, these oxidative and proinflammatory responses. These protective effects of nicotine were fully reversed by inhibition of neuronal nicotinic receptors by mecamylamine. The present results indicate that nicotine administration can attenuate the oxidative injury to spinal cords and suggest that neuronal nicotinic receptors can be attractive targets for neuroprotective therapy. PMID- 15135228 TI - Over expression of ATF-3 protects rat hippocampal neurons from in vivo injection of kainic acid. AB - ATF-3 is a member of the ATF superfamily of transcription factors and is strongly associated with episodes of cellular stress. We demonstrate an association between increases in ATF-3 protein and resistance to exitotoxic cell death in vivo. Intra-hippocampal injection of kainic acid elicited a robust increase in endogenous ATF-3 within kainate-resistant cells of the dentate gyrus, while overexpression of exogenous ATF-3 was found to protect vulnerable CA3 neurons from the same insult. These results suggest a positive contribution to neuronal survival in the context of stress-induced death, and support an anti-apoptotic role for ATF-3 in the brain. PMID- 15135229 TI - Quantitative comparison of placental expression of three aquaporin genes. AB - Three water channel proteins, aquaporins, have been shown to be expressed in the placentae of humans and sheep-AQP1, 3, 8; AQP1 is in the vasculature, whereas AQP3, 8 are in the trophoblast cells. In this study we used the sensitive and reproducible technique of real-time PCR to compare the level of expression of the mRNAs for AQP1, 3 and 8 in the ovine placenta at five stages of gestation (27, 45, 66, 100 and 140 days-where term approximately 150 days). AQP3 was quantitatively the most highly expressed AQP at 66, 100, and 140 days). At 27 days before significant trophoblast development had occurred, the only AQP present was AQP1, in the vasculature. The expression of these aquaporins underlie the high water and urea permeability of the ovine placenta in the last half of pregnancy. PMID- 15135230 TI - An analysis using DNA microarray of the time course of gene expression during syncytialization of a human placental cell line (BeWo). AB - Placental trophoblast syncytialization is a unique biological process. We have studied the time course of this process using DNA microarray in a cell model of syncytialization (the cytotrophoblast cell line BeWo following increased intracellular cAMP by forskolin). Total RNA was extracted from BeWo cells and labelled-cRNA target was then hybridized to a specific oligonucleotide probe set containing probes to over 12?000 human transcripts. Detectable levels of signal were found on average for 44 per cent of the total number of genes assayed. The correlation coefficient for the level of expression of independent replicates was #10878;0.99. The mRNA expression profile of specific genes analysed by microarray correlated quantitatively well with that analysed by reverse transcription polymerase chain reaction and with protein secretion. In the absence of forskolin there are relatively few changes in gene expression (reaching a threshold of two fold); in the presence of forskolin there are a substantial number of changes. By clustering the patterns of altered gene expression at least ten groups could be extracted. Seven of these clusters involved increased gene expression and three decreased expression. Each cluster has been categorized by gene ontology (confining the analysis to genes with 'known' function). Among the genes with increased expression following forskolin treatment were many required for cellular communication (such as placental specific peptide hormones) and metabolism (such as cholesterol side chain cleavage enzyme). Several genes known to be involved in cell adhesion and fusion have markedly changed expression levels very early following forskolin exposure, thus preceding morphological fusion of BeWo cells. Further analysis of this data and expression profiling in general will be able to contribute to understanding the functional basis for the formation of the placental syncytiotrophoblast. PMID- 15135231 TI - Distribution of glutamate transporters in the human placenta. AB - Glutamate metabolism is known to be important for growth and development of the human fetus. The glutamate transporters EAAT1, EAAT2 and EAAT3 are key components of the glutamate-glutamine cycle and responsible for active transport of glutamate over the cell membrane. The placenta is thought to regulate glutamate transport during fetal development. Glutamate transporters have been found in placentae of rats, but their distribution in the human placenta is unknown. Therefore, the distribution of glutamate transporters EAAT1, EAAT2 and EAAT3 were analysed in the human placenta during normal pregnancies ending between 8 and 40 weeks of gestation and in placentae of intrauterine growth restricted infants with gestational ages between 28 and 35 weeks of pregnancy. Using immunohistochemistry, EAAT1 expression was found in the syncytiotrophoblast layer, while EAAT2 was detected in the syncytiotrophoblast layer and in endothelial cells of about 5 per cent of all fetal blood vessels. EAAT3 was observed in the endothelium of the fetal blood vessels in all placentae examined. However, expression was also found in the syncytio- and the cytotrophoblast layer of the fetal villi at 8 weeks of gestational age. The expression patterns of EAAT1, EAAT2 and EAAT3 suggest involvement in active transport of glutamate between the fetal and maternal blood circulation. No differences were found in the distribution of the glutamate transporters between control and IUGR placentae. Our data show specific localization of EAAT1, EAAT2 and EAAT3 in the human placenta during development. PMID- 15135232 TI - Polarized lactate transporter activity and expression in the syncytiotrophoblast of the term human placenta. AB - We investigated the polarization of l-lactate transport in human syncytiotrophoblast by measuring uptake of [(14)C] l-lactate by both microvillous (maternal-facing; MVM) and basal (fetal-facing; BM) plasma membranes. [(14)C] l lactate uptake by MVM and BM was stimulated in the presence of an inwardly directed H(+)gradient, with a significantly higher uptake in MVM than in BM at initial rate (15.4+/-2.3 vs 5.6+/-0.6 pmol/mg protein/20 sec). Stereospecific inhibition was observed in MVM, with a higher affinity for l-lactate compared with d-lactate. In BM, there was no difference in the inhibition by these two stereoisomers. Inhibition of lactate uptake in both MVM and BM by 4,4' diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS), an inhibitor of monocarboxylate transporter (MCT) activity, indicated MCT-mediated mechanisms across both membranes. Kinetic modelling supported a two-transporter model as the best fit for both MVM and BM, the K(m)of the major component being 6.21 mm and 25.01 mm in MVM and BM respectively. Western blotting and immunolocalization examining the distribution of MCT1 and MCT4, showed that MCT expression was polarized, MCT1 being predominantly localized to BM and MCT4 showing greater abundance on MVM. CD147, a chaperone protein for MCT1 and MCT4, was equally expressed by both membranes. These studies demonstrate that the opposing plasma membranes of human syncytiotrophoblast are polarized with respect to both MCT activity and expression. PMID- 15135233 TI - Non-gastric H+/K+ ATPase is present in the microvillous membrane of the human placental syncytiotrophoblast. AB - In humans, the non-gastric H(+)/K(+)ATPase (ATP1AL1) has previously been shown to be expressed in the epithelia of skin, kidney and colon. In this study we tested the hypothesis that the non-gastric H(+)/K(+)ATPase is localized to the syncytiotrophoblast, the transporting epithelium of the human placenta. Microvillous (MVM) and basal plasma membranes (BM) of the syncytiotrophoblast were isolated from term placenta and membrane proteins were separated using SDS PAGE. The ATP1AL1 protein was identified as a 114 kD band in both MVM and BM by Western blot, however, the protein was more abundant in the MVM. Using immunocytochemistry H(+)/K(+)ATPase protein was localized in MVM but not BM. We constructed primers specific for ATP1AL1 and performed RT-PCR on RNA isolated from human placenta and human kidney. A product of the expected size could be detected in both tissues after 30 cycles of amplification. The sequence identity of this 517 nucleotide product was confirmed by sequencing and found to be identical to the human non-gastric H(+)/K(+)ATPase. The activity of this proton pump appears to be low in normal healthy placental at term, however, it is speculated that MVM non-gastric H(+)/K(+)ATPase may be important in pathological states. In conclusion, non-gastric H(+)/K(+)ATPase is present in the microvillous plasma membrane of the transporting epithelia of the human placenta. PMID- 15135234 TI - Expression and localization of menkes and Wilson copper transporting ATPases in human placenta. AB - Copper is an essential trace element necessary for normal growth and development. During pregnancy, copper is transported from the maternal circulation to the fetus by mechanisms which have not been clearly elucidated. Two copper transporting ATPases, Menkes (ATP7A; MNK) and Wilson (ATP7B; WND) are known to be expressed in the placenta and are thought to have a role in copper transport to the fetus. In this study, the expression and localization of the MNK and WND proteins in the human placenta were investigated in detail using immunoperoxidase and double-label immunohistochemistry. MNK and WND are differentially localized within the placenta. MNK is present in the syncytiotrophoblast, the cytotrophoblast and the fetal vascular endothelial cells whereas WND is only in the syncytiotrophoblast. Placental levels of both proteins, measured by Western blot analysis, did not change across pregnancy. These data offer some insights into possible roles for MNK and WND within the placenta. PMID- 15135235 TI - Norepinephrine transporter (NET), serotonin transporter (SERT), vesicular monoamine transporter (VMAT2) and organic cation transporters (OCT1, 2 and EMT) in human placenta from pre-eclamptic and normotensive pregnancies. AB - Pre-eclampsia is one of the most common causes of perinatal and maternal morbidity and mortality. High blood pressure and proteinuria are important clinical signs of pre-eclampsia. Sympathetic overactivity and elevated level of circulating vaso active substances, such as monoamines has been shown. Extracellular concentrations of monoamines are normally kept low by specific transporter proteins of which many are expressed in the placenta. In this study we used in situ hybridization and real-time PCR to study the gene expression of monoamine transporters, such as NET, SERT, VMAT2, EMT and OCT1/2, in normal as well as in pre-eclamptic placentae. We demonstrated high expression of NET mRNA in the trophoblast cells of the anchoring villi and a lower expression intensity in the chorionic villi. SERT mRNA was mainly detected in chorionic villi. VMAT2 mRNA was not detected in the central part of the placenta but was present in the spiral arteries of placenta bed biopsies, in cytokeratin positive cells. EMT mRNA was mainly detected in the intra lobular septa and together with OCT1 and OCT2 mRNAs also expressed in scattered cells of placental vessel adventitias. Moreover, quantitative analysis showed a significant lower expression of NET and EMT mRNAs in pre-eclamptic placentae as compared to the control group. A defective gene expression or function of these monoamines transporters might explain the elevated concentrations of monoamines in pre-eclamptic patients. Monoamine transporters may serve as a protective mechanism preventing vasoconstriction in the placental vascular bed and thereby securing a stable blood flow to the fetus. PMID- 15135236 TI - Expression of Smad2 and Smad4 in mouse uterus during the oestrous cycle and early pregnancy. AB - SMAD2 and SMAD4 are intracellular transducers of TGF-beta superfamily. In situ hybridization and semi-quantitative RT-PCR were employed to determine the temporal and spatial expression of Smad2 and Smad4 mRNA in mouse uterus during the oestrous cycle and early pregnancy. Smad2 mRNA was predominantly present in the luminal and glandular epithelium at dioestrus and prooestrus, while Smad4 expression was at a steady level in the luminal and glandular epithelium throughout the oestrous cycle. During pre-implantation period, Smad2 hybridization signals were accumulated in the luminal and glandular epithelium at a basal level; Smad4 mRNA appeared in the epithelium with a little variation in hybridization signal intensity. After implantation, on day 5 of pregnancy, Smad2 signals were localized to the subluminal stroma surrounding the implanting blastocyst, and Smad4 mRNA were accumulated in the decidua near the luminal epithelium. Both Smads were present in the decidua on days 6-7 with a switch from the mesometrial pole to the antimesometrial pole. RT-PCR results showed that both Smad2 and Smad4 mRNA levels were rising during peri-implantatation. The results suggest that Smad2 and Smad4 might be involved in the cycling changes of mouse uterus during the oestrous cycle and embryo implantation. PMID- 15135237 TI - Patterns of uterine cellular proliferation and apoptosis in the implantation site of the rat during pregnancy. AB - During gestation, the balance between cell proliferation and death is crucial for successful embryo implantation and maintenance of pregnancy. The uterine endometrium responds to blastocyst implantation with extensive proliferation and differentiation of stromal cells into decidual cells, forming the antimesometrial and mesometrial decidua, which regress by apoptosis. In the latter region it is also observed the growth of metrial gland. To elucidate the events underlying this tissue remodelling we investigated the spatial and temporal pattern of expression of the proliferating cell nuclear antigen (PCNA) and localized the apoptotic cells, by the TUNEL assay and by the expression of active caspase-3. We found that PCNA is expressed at high levels during decidualization until day 12 of gestation declining thereafter abruptly. On the contrary, the appearance of apoptotic cells was detected, by the TUNEL and active caspase-3 expression, in the mesometrial decidua on day 12, increasing from days 14 to 16 in the decidua and metrial gland. In the antimesometrial decidua apoptosis was observed from early to day 12 of pregnancy. However, on day 13 only cell debris and neutrophils were observed, indicating also the presence of necrosis. These results suggest that decidual cells undergo, in distinct regions and at different stages of pregnancy, cell death by apoptosis and secondary necrosis. PMID- 15135238 TI - Androgen secretion by Rcho-1 cells is independent of extracellular glutamate concentration. AB - In primates, progesterone, secreted by the placenta, is important for the maintenance of pregnancy. Androstenedione, a progesterone metabolite, fulfils a similar role in the rodent. Prior work has suggested that glutamate sufficiency and subsequent oxidation is important to placental androgen synthesis, presumably because of the production of NADPH (Trophoblast Res (1993) 7, 77). Rcho-1 cells possess a phenotype similar to that of rat placental giant cells, and secrete androstenedione and progesterone when in the differentiated state (J Endocrinol (1996) 150, 161). Our objective was to determine whether extracellular glutamate concentrations impact hormone synthesis in Rcho-1 cells. Rcho-1 cells were kept in culture under differentiating conditions. Extracellular glutamate concentrations were varied from 0-5 mm, and hormone concentrations assayed by ELISA. Rcho-1 cells secreted both progesterone and androstenedione. There was no direct correlation between the extracellular concentration of glutamate and the secretion of either hormone. Inhibition of transaminases (aminooxyacetic acid) or of glutaminase (6-diazo-5-oxo-l-norleucine) did not alter hormone production. Therefore, extracellular glutamate concentrations did not impact progesterone or androstenedione secretion. These findings may relate to the central position of glutamate in a variety of metabolic pathways, making intracellular depletion of this amino acid difficult to accomplish, or may represent a species specific difference in regulation. PMID- 15135239 TI - Complex patterns of GCM1 mRNA and protein in villous and extravillous trophoblast cells of the human placenta. AB - The Gcm1 gene encodes a transcription factor that is essential for both syncytiotrophoblast differentiation and formation of chorionic villi in mice. Its early expression is very unusual in that it defines a subset of trophoblast cells in the chorion, a layer that otherwise contains trophoblast stem cells. While Gcm1 mRNA expression initiates independently within the chorion, the subsequent maintenance of mRNA expression as well as the onset of protein accumulation is dependent on contact with allantoic mesoderm. Previous studies have shown that human GCM1 mRNA and protein are detectable in the placenta, but their patterns have not been compared nor precisely localized. We, therefore, conducted the present study to determine if the human mRNA and protein are subject to the same complexities of regulation as the mouse. In situ hybridization studies showed that the GCM1 mRNA was expressed in villous cytotrophoblast cells, but only a subset and never within cells immediately at the base of columns. Interestingly, the mRNA was detected throughout the cytotrophoblast columns. GCM1 protein expression studies demonstrated that the transcription factor was present mainly within the nuclei of a subset of cytotrophoblast cells, consistent with its role as a transcription factor. Feint cytoplasmic staining of the transcription factor was found in the syncytiotrophoblast but not in aggregated syncytial nuclei. Nuclear immuno-reactivity for the GCM1 protein was detected in occasional nuclei in the distal part of the column. Therefore, GCM1 expression is regulated both at the transcriptional and translational level. Overall, these studies show that the general features of GCM1 mRNA and protein expression in the human placenta are conserved with the mouse. They also highlight the fact that villous cytotrophoblast cells are extremely heterogeneous with respect to GCM1 expression, a factor that should be considered when using isolated cytotrophoblast cells for culture studies. PMID- 15135240 TI - Sequential expression of VEGF and its receptors in human placental villi during very early pregnancy: differences between placental vasculogenesis and angiogenesis. AB - Vascularization within the human placenta is the result of the de novo formation of vessels derived from pluripotent precursor cells in the mesenchymal core of the villi. Vascularization of placental villi starts at around day 21 post conception (p.c.) with a four somite embryo. At this stage progenitors of haemangiogenic cells differentiate to form first vessels. These progenitor cells are thought to be directly derived from mesenchymal cells rather than originating from fetal blood cells. We investigated the relation between differentiation of stromal cells towards endothelial cells and vascular structures and the expression pattern of the respective growth factors. Using transmission electron microscopy and immunohistochemistry (for VEGF, Flt-1, Flk-1, CD14, CD34, and CD68) the development of placental vasculogenesis during very early stages of pregnancy (days 22-48 p.c.) was studied. We found that VEGF is strongly expressed in villous cytotrophoblast cells and subsequently in Hofbauer cells while its receptors Flt-1 and Flk-1 are found on vasculogenic and angiogenic precursor cells. The developmental expression and secretion of VEGF suggests its involvement in recruitment, maintenance and formation of first angiogenic cells and vessels. Interactions between VEGF and Flk-1 and Flt-1 may regulate placental vasculogenesis and angiogenesis in a paracrine and autocrine manner. The sequential expression of growth factors in different cell types may point to the fact that placental vasculogenesis and angiogenesis are clearly distinct events. PMID- 15135241 TI - Vitamin C exacerbates hydrogen peroxide induced apoptosis and concomitant PGE2 release in amnion epithelial and mesenchymal cells, and in intact amnion. AB - This study was undertaken to investigate the effect of hydrogen peroxide (HP), a reactive oxygen species, and vitamin C, an antioxidant, on apoptosis and prostaglandin (PGE(2)) release in human amnion epithelial and mesenchymal cells, and intact amnion. Amnion cells and explants were incubated with and without HP and vitamin C. Cytoproliferation assay for viability, DNA fragmentation and PARP cleavage for apoptosis, EIA for PGE(2), and western blots for cyclooxygenases (COX) were performed. In amnion cells and explants, HP (0-5 mm) induced dose dependent apoptosis as per DNA fragmentation and PARP cleavage. HP (0-0.5 mm) also induced PGE(2)release concomitant with apoptosis in both cell types. In amnion explants, HP (0-10 mm) induced COX-2 protein and PGE(2)release concomitant with apoptosis. Vitamin C (0.01-10 mm), alone, enhanced epithelial but inhibited mesenchymal cell viability. It induced PGE(2)release in amnion explants. Vitamin C (1 mm) failed to inhibit HP induced apoptosis, but instead exacerbated it in epithelial and mesenchymal cells, and amnion explants. Vitamin C (0-10 mm) enhanced HP induced PGE(2)in mesenchymal cells. HP induces concomitant apoptosis and PGE(2)release in amnion epithelial and mesenchymal cells, and in intact amnion explants. HP induced apoptosis is not inhibited but enhanced by vitamin C. PMID- 15135242 TI - A simple method for comparing immunogold distributions in two or more experimental groups illustrated using GLUT1 labelling of isolated trophoblast cells. AB - Colloidal gold-labelling, combined with transmission electron microscopy, is a valuable technique for high-resolution immunolocalization of identified antigens in different subcellular compartments. Whilst the technique has been applied to placental tissues, few quantitative studies have been made. Subcellular compartments exist in three main categories (viz. organelles, membranes, filaments/tubules) and this affects the possibilities for quantification. Generally, gold particles are counted in order to compare either (a) compartments within an experimental group or (b) compartmental labelling distributions between groups. For the former, recent developments make it possible to test whether or not there is differential (nonrandom) labelling of compartments. The methods (relative labelling index and labelling density) are ideally suited to analysing label in one category of compartment (organelle or membrane or filament) but may be adapted to deal with a mixture of categories. They also require information about compartment size (e.g. profile area or trace length). Here, a simple and efficient method for drawing between-group comparisons of labelling distributions is presented. The method does not require information about compartment size or specimen magnification. It relies on multistage random sampling of specimens and unbiased counting of gold particles associated with different compartments. Distributions of observed gold counts in different experimental groups are compared by contingency table analysis with degrees of freedom for chi-squared (chi(2)) values being determined by the numbers of compartments and experimental groups. Compartmental values of chi(2)which contribute substantially to total chi(2)identify the principal subcellular sites of between-group differences. The method is illustrated using datasets from immunolabelling studies on the localization of GLUT1 glucose transporters in cultured human trophoblast cells exposed to different treatments. PMID- 15135244 TI - HIV-related lipodystrophy and related factors. AB - As new therapies for HIV infection have been developed, some of the clinical focus related to AIDS and HIV infection has shifted from acute care, to more chronic issues. Some of these new clinical issues seem related to the HIV infection itself, while others seem to be side effects of therapeutic efforts. Metabolic abnormalities, such as dyslipidemia, insulin resistance, and lipodystrophy (LD) have been observed. The clinical importance of these is demonstrated by the increased prevalence of cardiovascular disease and diabetes in HIV infected persons. LD is a general term used to describe varying degrees of fat redistribution, including lipoatrophy and lipohypertrophy, in different body regions. Though LD was observed in persons with HIV infection before highly active treatment regimens were developed, the prevalence of LD has seemingly increased drastically with the widespread use of more active therapies. It has been postulated that protease inhibitors (PI), especially, are linked to the development of LD. This review will assess the epidemiologic information related to HIV-associated LD, and related metabolic syndromes. In addition, potential mechanisms accounting for these syndromes will be reviewed. In general, the available data do not define a single, definable etiology or mechanism explaining these clinical conditions, but suggest that these conditions are caused by a complex interaction potentially involving such things as the side effects of medications, alteration of immune function, and individual subject characteristics, such as body weight and baseline lipid level. PMID- 15135243 TI - Placental folate transport and binding are not impaired in pregnancies complicated by fetal growth restriction. AB - Maternal folate deficiency is associated with fetal growth restriction, however, transfer of folate across placentae of pregnancies complicated by fetal growth restriction has never been investigated. We studied whether maternal to fetal 5 methyltetrahydrofolate (5MTF) transport in the ex vivo dually perfused isolated cotyledon, binding of [(3)H] folate (PteGlu) to the syncytial microvillous membrane, and protein expression of folate receptor alpha (FR-alpha) and reduced folate carrier (RFC) in these placentae are disturbed. Placental clearance of 5MTF from the maternal perfusate appeared to be non-saturable over a range of 50 to 500 nm, independent of albumin and flow-independent. No statistically significant differences between placentae complicated with fetal growth restriction and uncomplicated pregnancies were observed. Binding characteristics of [(3)H-]PteGlu to microvillous membranes of fetal growth restriction versus control placentae were similar: B(max)of 3.9+/-2.0 (mean+/-s.d.) versus 4.0+/-1.6 pmol/mg protein and a K(d)of 0.037+/-0.010 versus 0.040+/-0.018 nm. Expression of FR-alpha and RFC were not different in placentae of both groups studied. In conclusion, fetal growth restriction appears not to be associated with impaired maternal to fetal placental folate transport, placental receptor binding, or expression of FR-alpha and RFC. PMID- 15135245 TI - A new but frequent mutation of apoB-100-apoB His3543Tyr. AB - ApolipoproteinB 100 (apoB-100) is an important component of atherogenic lipoproteins such as LDL and serves as a ligand for the LDL-receptor. Familial defective apolipoproteinB 100 (FDB) is caused by a R3500Q mutation of the apoB gene and results in decreased binding of LDL to the LDL-receptor. So far FDB is the most frequent and best studied alteration of apoB-100. Apart from this, three other apoB mutations, R3500W, R3531C and R3480W, affecting binding to the LDL receptor are known to date. We screened the apoB gene segment of codons 3448-3561 by denaturing gradient gel electrophoresis (DGGE) analysis in a total of 853 consecutively sampled German patients undergoing diagnostic coronary angiography for suspected CAD. By this, a new single base mutation was detected and confirmed by DNA sequencing. The mutation, CAC(3543)TAC results in a His3543Tyr substitution in apoB-100 (H3543Y). The prevalence of heterozygotes for H3543Y in the study population was 0.47% compared to 0.12% for the known Arg 3500 Gln (R3500Q) mutation. In conclusion, the new mutation is four times more frequent than "classical" FDB and thus appears to be the most common apoB mutation in Germany. PMID- 15135246 TI - Stimulation of Na-dependent phosphate transport by platelet-derived growth factor in rat aortic smooth muscle cells. AB - We investigated the effect of platelet-derived growth factor B homodimer (PDGF BB) on inorganic phosphate (Pi) transport activity, which has been reported to be involved in the mechanism of atherosclerosis, in A-10 rat aortic vascular smooth muscle cells (VSMCs). PDGF-BB time- and dose-dependently stimulated Pi transport in A-10 cells. Using northern blot analysis, the PDGF-BB-enhanced Pi transporter (PiT) in A-10 cells was identified as Pit-1 (Glvr-1), a member of the type III Na dependent PiT. An inhibitor of PDGF beta-receptor tyrosine kinase suppressed PDGF BB-induced Pi transport. Both a protein kinase C (PKC) inhibitor calphostin C and PKC down regulation suppressed the stimulatory effect of PDGF-BB on Pi transport. On the other hand, inhibition of mitogen-activated protein (MAP) kinases by selective inhibitors did not affect Pi transport. Ly294002, a phosphatidylinositol (PI) 3-kinase inhibitor, partially attenuated PDGF-BB induced Pi transport. A selective inhibitor of S(6) kinase, rapamycin, reduced this effect of PDGF-BB, while Akt kinase inhibitor did not. In summary, these results indicated that PDGF-BB is a potent and selective stimulator of Pi transport in VSMCs. The mechanism responsible for this effect is not mediated by MAP kinase, but involves activation of PKC, PI 3-kinase and S(6) kinase. PMID- 15135247 TI - Evaluation of a soft atherosclerotic lesion in the rabbit aorta by an invasive IVUS method versus a non-invasive MRI technology. AB - The intravascular ultrasound (IVUS) modality has rapidly gained acceptance for the measurement of arterial plaque thickness and for anatomical characterization. In view, however, of the growing interest in the direct assessment of plaque size after therapeutic modalities directly reducing plaque burden, a non-invasive method such as magnetic resonance imaging (MRI) may be of help for repeated evaluations. The two methods were compared directly on a focal plaque developed at the abdominal aortic level by a combination of local electric lesion followed by a hypercholesterolemic diet. The plaque was fully characterized histopathologically at intervals up to 120 days from lesion induction, and maximal plaque formation was detected at 90 days from electrical injury. Plaques could be well assessed by IVUS at each time point analyzed and data correlated very well to histopathologic findings (r = 0.969, P = 0.0014). The MRI technology provided reliable determinations only at 90 days after lesion induction, i.e. at maximal plaque formation, with excellent correspondence to IVUS determinations (r = 0.989, P = 0.0111). Altogether these findings indicate that the non-invasive MRI technology, when applied to the analysis of arterial plaques of adequate size, can be used successfully for plaque determination, with results comparable to the invasive IVUS technique. PMID- 15135248 TI - Involvement of leukotactin-1, a novel CC chemokine, in human atherosclerosis. AB - Atherosclerosis is a chronic inflammatory disease that involves the recruitment of leukocytes to the arterial wall. Leukotactin-1 (Lkn-1), a new member of the CC chemokine family, is a potent chemoattractant for leukocytes and thus is implicated in inflammatory diseases. In this study, we investigated the possible association of Lkn-1 with human atherosclerosis. Atherosclerotic lesion and plasma samples were obtained from atherosclerotic patients. Human THP-1 monocyte derived foam cells were prepared by treatment with an oxidized low-density lipoprotein. The expression level of Lkn-1 or its receptors mRNA was measured by RT-PCR analysis. Levels of plasma Lkn-1, MCP-1, ICAM-1 and total cholesterol were measured by ELISA or enzymatic assay. Lkn-1 expression was markedly enhanced not only at the mRNA level in human atherosclerotic lesions, but also at the circulating level in atherosclerosis patients compared with normal subjects. An in vitro study revealed that the level of Lkn-1 release was significantly enhanced by oxidative stress or proatherogenic mediators in macrophages and macrophage-derived foam cells. Lkn-1 stimulated endothelial cells to release ICAM 1, which is implicated in atherogenesis. Taken together, our data suggest that Lkn-1 plays an important role in the development of atherosclerosis. PMID- 15135249 TI - Methionine synthase reductase MTRR 66A > G has no effect on total homocysteine, folate, and Vitamin B12 concentrations in renal transplant patients. AB - The association of variants of the gene encoding methionine synthase reductase (MTRR) with hyperhomocysteinemia, folate and Vitamin B(12) status in kidney graft recipients is unknown. We examined two mutations in MTRR in a cross-sectional study of 733 kidney graft recipients. The allele frequency of MTRR 66G was 0.55. 369 patients (50.3%) were heterozygous and 219 patients (29.9%) were homozygous for the mutation. None of the patients showed the 997C > G mutation. The allelic variants of MTRR 66A > G showed no significant association with total homocysteine (tHcy) levels, both in univariate analyses, and in a multivariate model controlling for age, gender, body mass index, renal function, time since transplantation, underlying kidney disease, as well as the MTHFR 677C > T/1298A > C genotypes. Similarly, no significant associations between the MTRR 66A > Ggenotypes and plasma folate or Vitamin B(12) levels were found. In conclusion, MTRR 66A > G has no major effect on tHcy, folate, or Vitamin B(12) plasma concentrations in kidney graft recipients. PMID- 15135250 TI - Type 2 diabetes is associated with impaired endothelium-dependent, flow-mediated dilation, but impaired glucose metabolism is not; The Hoorn Study. AB - BACKGROUND: Type 2 diabetes (DM2) and impaired glucose metabolism (IGM) are associated with an increased cardiovascular disease risk. Impaired endothelial synthesis of nitric oxide (NO) is an important feature of atherothrombosis and can be estimated from endothelium-dependent flow-mediated dilation (FMD). It is controversial whether or not FMD is impaired in DM2 and IGM. We investigated this issue in a population-based setting. METHODS AND RESULTS: In the study population (n = 650; 246 with normal glucose metabolism (NGM), 135 with IGM and 269 with DM2; mean age: 67.6 years), FMD and endothelium-independent nitroglycerine mediated dilation (NMD) were ultrasonically estimated from the brachial artery and expressed as the absolute change in diameter in mm. The increase in diameter (mean +/- standard deviation) in NGM, IGM and DM2 was 0.19 +/- 0.15, 0.19 +/- 0.18 and 0.13 +/- 0.17 MD and 0.45 +/- 0.21, 0.43 +/- 0.24 and 0.45 +/- 0.25 for NMD. After adjustment for age, sex, baseline diameter and percentage increase in peak systolic velocity, DM2, as compared to NGM, remained associated with impaired FMD (regression coefficient beta (95%CI)) as compared to NGM, -0.06 mm ( 0.09 to -0.03). IGM was not associated with impaired FMD (beta, 0.01 mm (-0.02 to 0.04)). Additional adjustment for conventional cardiovascular risk factors did not alter these associations. Hyperglycemia or hyperinsulinemia explained 2% of the association between DM2 and FMD. NMD was not associated with glucose tolerance. CONCLUSIONS: This study shows that DM2 is independently associated with impaired FMD. Hyperglycemia and hyperinsulinemia contribute minimally to this association. Impaired FMD may therefore, in part, explain the increased cardiovascular disease risk in DM2, whereas the normal FMD in IGM suggests that other forms of endothelial dysfunction are important in explaining the increased cardiovascular disease risk in IGM. PMID- 15135251 TI - Genetic polymorphisms affecting the phenotypic expression of familial hypercholesterolemia. AB - The clinical expression of heterozygous familial hypercholesterolemia (FH) is highly variable even in patients carrying the same LDL receptor (LDL-R) gene mutation. This variability might be due to environmental factors as well as to modifying genes affecting lipoprotein metabolism. We investigated Apo E (2, 3, 4), MTP (-493G/T), Apo B (-516C/T), Apo A-V (-1131T/C), HL (-514C/T and -250G/A), FABP-2 (A54T), LPL (D9N, N291S, S447X) and ABCA1 (R219K) polymorphisms in 221 unrelated FH index cases and 349 FH relatives with defined LDL-R gene mutations. We found a significant and independent effect of the following polymorphisms on: (i) plasma LDL-C (Apo E, MTP and Apo B); (ii) plasma HDL-C (HL, FABP-2 and LPL S447X); (iii) plasma triglycerides (Apo E and Apo A-V). In subjects with coronary artery disease (CAD+), the prevalence of FABP-2 54TT genotype was higher (16.5% versus 5.2%) and that of ABCA1 219RK and KK genotypes lower (33.0% versus 51.5%) than in subjects with no CAD. Independent predictors of increased risk of CAD were male sex, age, arterial hypertension, LDL-C level and FABP-2 54TT genotype, and of decreased risk the 219RK and KK genotypes of ABCA1. These findings show that several common genetic variants influence the lipid phenotype and the CAD risk in FH heterozygotes. PMID- 15135252 TI - Genetic diagnosis of familial hypercholesterolaemia: a mutation and a rare non pathogenic amino acid variant in the same family. AB - Familial hypercholesterolaemia (FH), a relatively common inherited disorder, is caused by mutations in the gene for the low density lipoprotein (LDL) receptor (LDLR) that result in impaired clearance of LDL. Identification of mutations in patients with the clinical phenotype of FH allows unequivocal diagnosis in potentially affected relatives, but depends critically on distinguishing mutations that affect protein function from variants with no significant effect. A presumed functional mutation in LDLR (G198D in exon 4) was identified in two hypercholesterolaemic English brothers by high throughput screening and was not found in 550 controls. However, a second variant (L458P) was identified separately in their mother that co-segregated with hypercholesterolaemia in the entire pedigree. L458, but not G198, is strongly conserved between species and lies in a region important for beta-propeller stability. G198D was inherited from their normolipidaemic father by two of three siblings heterozygous for L458P; they appeared less severely hypercholesterolaemic and more responsive to statins than the third affected brother and their mother. This study emphasises that apparent co-segregation of an amino acid substitution in a critical region of the protein with hypercholesterolaemia and its absence from a large control population is insufficient evidence that a variant of the LDL receptor is necessarily deleterious to its function. PMID- 15135253 TI - Influence of the -514C/T polymorphism in the promoter of the hepatic lipase gene on postprandial lipoprotein metabolism. AB - The -514C/T polymorphism located in the promoter region of the hepatic lipase gene mediates changes in the plasma levels of the enzyme. The aim of this study was to determine whether the presence of this polymorphism modifies the postprandial clearance of lipoproteins of intestinal origin. 51 normolipemic volunteers, homozygotes for the allele E3 of the apo E were selected (26 homozygotes for the C allele and 25 carriers of the T allele in both homozygote and heterozygote form). The subjects underwent a Vitamin A fat-loading test. Blood was drawn every hour until the 6th hour and every 2 h and 30 min until the 11th hour to determine cholesterol and plasma triglycerides as well as cholesterol, triglycerides (TG) and retinyl palmitate in triacylglycerol-rich lipoproteins (chylomicrons and chylomicron remnants). Carriers of the T allele showed significantly lower postprandial levels of apolipoprotein B (P < 0.01), total TG in plasma (P < 0.05), small TRL-TG (P < 0.04), large TRL-TG (P < 0.04) and small TRL-cholesterol (P < 0.04) when compared to subjects homozygous for the C allele. Our data suggest that the T allele of the -514C/T polymorphism in the promoter region of the hepatic lipase gene is associated with a lower postprandial lipemic response. PMID- 15135254 TI - The -344T>C promoter variant of the gene for aldosterone synthase (CYP11B2) is not associated with cardiovascular risk in a prospective study of UK healthy men. AB - INTRODUCTION: The tissue renin-angiotensin system is implicated in the pathogenesis of coronary artery disease (CAD). As locally synthesised aldosterone is a potential mediator of CAD, we have sought an association of the -344T>C variant of the aldosterone synthase (CYP11B2) gene with CAD events. METHODS: Subjects comprised of the Second Northwick Park Heart Study (NPHSII), a prospective study of unrelated, healthy middle-aged Caucasian males. CAD events were recorded in 2490 subjects, and defined as a sudden cardiac death, myocardial infarction or coronary artery revascularisation procedure. Mean follow-up was 10.8 years. Aldosterone synthase genotype was determined in 2490 subjects. Power calculation suggests that we have 80% power (at a significance level of 0.05) to detect a difference in hazard ratio (HR) between homozygote groups of 0.45. RESULTS: One hundred and eighty-seven CAD events were recorded in 2490 subjects. In the group overall, CAD events were independent of genotype with adjusted hazard ratios being 1.00 versus 1.25 versus 0.80 for TT versus TC versus CC genotypes, respectively, P = 0.07. Genotype interactions with smoking and blood pressure were sought. Whilst CAD events were independent of genotype amongst non smokers, CC genotype in smokers was associated with a reduced risk HR 2.02 versus 2.28 versus 0.82 for TT versus TC versus CC genotypes, P = 0.05 (HR for TT + TC versus CC were 1.77 versus 0.67, P = 0.02). This apparent interaction remained after adjustment for conventional risk factors. No such interaction was found with blood pressure. CONCLUSIONS: Aldosterone synthase genotype is unrelated to overall CAD events risk. A possible interaction with smoking requires confirmation. PMID- 15135255 TI - Elevated levels of plasma homocysteine are associated with neurotoxicity. AB - OBJECTIVE: The aim of the present study is to investigate whether increased plasma homocysteine (tHcy) has an effect on the cerebral metabolic concentrations, reflecting neurotoxicity, measured with Magnetic Resonance Spectroscopy (MRS). METHODS: One hundred and thirteen patients had a MRS investigation of the brain and the concentration of N-acetyl-aspartate (NAA), choline and creatine was measured and fasting plasma tHcy was assessed. We used linear regression models to investigate the association between tHcy and cerebral metabolic concentrations. RESULTS: We found that tHcy is associated with cerebral NAA (B = -0.09 mmol/l (95% CI, -0.17 to -0.00)) and with creatine (B = -0.09 mmol/l (95% CI, -0.16 to -0.02)) per 1 micromol/l increase tHcy. Patients with a tHcy >/=14 micromol/l had a lower concentration cerebral of NAA (P < 0.05) and creatine (P < 0.01) compared with patients with a tHcy <14 micromol/l. No significant association was found for the cerebral choline. CONCLUSION: Elevated levels of tHcy were associated with low concentrations of cerebral NAA and creatine, independent of renal function or the presence of atherosclerotic disease. These preliminary results suggest that tHcy has a neurotoxic effect in vivo. PMID- 15135257 TI - Hepatitis B and C virus infection and the risk of atherosclerosis in a general population. AB - BACKGROUND: The development of atherosclerosis has an inflammatory component. Currently it is not clear, whether hepatitis B and C virus infections are associated with the risk of atherosclerosis. The aim of the present analysis was to investigate those relationships in a population sample. METHODS AND RESULTS: The study of health in Pomerania (SHIP) is a cross-sectional study of the adult population in the northeast of Germany. HBs antigen (HBsAg) and IgG antibodies against hepatitis B and C virus (anti-HBs and anti-HCV) were determined by enzyme linked immunosorbent assays. Fifteen subjects (0.4%) were positive for HBsAg, and 21 subjects (0.5%) were positive for anti-HCV. Among the persons who had no history of anti-hepatitis B vaccination, 213 individuals (5.0%) were found to be as positive for anti-HBs. These individuals and those with prevalent anti-HCV antibodies were regarded as cases (n = 233). The control group comprised of 4033 individuals. Multivariable analyses revealed that there was no independent association between anti-HBs and anti-HCV antibody seropositivity and atherosclerotic end-points such as prevalent myocardial infarction, stroke, carotid intima-media thickness (IMT), carotid plaques and stenoses. CONCLUSION: There is no association between serological markers for hepatitis B and C virus infection and the risk of atherosclerosis in this population sample. PMID- 15135256 TI - Rosuvastatin reduces MMP-7 secretion by human monocyte-derived macrophages: potential relevance to atherosclerotic plaque stability. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) have been shown to reduce cardiovascular morbidity and mortality by their actions on atherogenic lipid profiles and by pleiotropic effects. In this study, we have investigated the effect of a new statin, rosuvastatin (Crestor), on sterol synthesis and the expression of metalloproteinases (MMPs) in human monocyte derived macrophages (HMDM). Rosuvastatin dose-dependently inhibited sterol synthesis from acetate with an IC(50) of 70 nM. In addition, MMP-7 levels were reduced in a dose-dependent manner with maximal inhibition of 50% (P < 0.01) at 1 microM. Also, addition of isoprenoids such as farnesyl pyrophosphate (Fpp) or geranylgeranyl pyrophosphate (GGpp) fully overcame the inhibitory effect of rosuvastatin on MMP-7. Neither quantitative PCR nor transient transfection of HMDM with a luciferase reporter construct under the control of human MMP-7 promoter (2300 bp of the 5' region on MMP-7 gene) showed a decrease in MMP-7 mRNA following treatment with rosuvastatin (10(-6)M). However, the inhibitory effect of the statin occurred at the post-transcriptional level as determined by actinomycin D experiment. In conclusion, several studies have reported a high expression of active MMP-7 in human atherosclerotic plaques indicating a potential role in the weakening of the fibrous cap, predisposing it to rupture. The effect of rosuvastatin in reducing MMP-7 might protect fibrous caps from degradation and in turn stabilize atheromatous plaques. PMID- 15135258 TI - Value of serum ferritin and soluble transferrin receptor for prediction of coronary artery disease and its clinical presentations. AB - We investigated whether there is an association between serum ferritin or soluble transferrin receptor (sTfR) concentrations and coronary artery disease (CAD) or its clinical presentations. This is a case-control study that included 892 patients (664 cases with angiographically proven CAD and 228 controls without CAD). Blood was collected before angiography for determination of sTfR, ferritin and C-reactive protein (CRP). The values (median, 25th-75th percentiles) of sTfR (2.6 [2.1; 3.2]mg/l versus 2.4 [2.1; 3.0]mg/l, P = 0.13) or ferritin (140.1 [74.8; 248.3]ng/ml versus 120.1 [74.9; 218.0]ng/ml, P = 0.11) did not differ significantly between cases or controls. The values of sTfR in the case subjects with 1-vessel, 2-vessel, and 3-vessel CAD were: 2.4 [2.0; 3.0], 2.6 [2.0; 3.2], and 2.8 [2.2; 3.3]mg/l, respectively (P = 0.003). In multivariate analysis, neither sTfR (chi2 = 0.14, P = 0.70) nor ferritin (chi2 = 2.8, P = 0.09) correlated independently with the presence of CAD. In case subjects with stable angina, unstable angina, and acute myocardial infarction (MI), ferritin concentrations were: 127.5 [69.5; 214.0], 138.9 [86.1; 278.0], and 175.0 [93.5; 314.5]ng/ml, respectively (P < 0.001). Our results showed that serum concentrations of sTfR or ferritin do not predict the risk for coronary artery disease. In subjects with pre-existing CAD, those with more severe disease had increased levels of sTfR. Patients with CAD presenting with acute coronary syndromes showed increased levels of serum ferritin. PMID- 15135259 TI - HIF-VEGF-VEGFR-2, TNF-alpha and IGF pathways are upregulated in critical human skeletal muscle ischemia as studied with DNA array. AB - Critical lower limb ischemia is a common cause for amputation. To develop new therapeutic strategies, more information is needed about molecular mechanisms of tissue responses to ischemic stress and factors inducing angiogenesis. Using a DNA array of 8400 genes, gene expression patterns in human skeletal muscle samples collected from lower limbs amputated due to acute-on-chronic or chronic critical lower limb ischemia, were compared with the control samples collected from the same limb. The results were confirmed by RT-PCR and immunohistochemistry. In acute-on-chronic ischemia, 291 genes were significantly upregulated and 174 genes were downregulated (change in 5.5% of all genes) as compared to control samples. Significant induction of the hypoxia-inducible angiogenic pathway involving hypoxia-inducible factor-1alpha (HIF-1alpha), HIF 2alpha, vascular endothelial growth factor (VEGF) and its angiogenic receptor VEGFR-2, as well as tumor necrosis factor-alpha (TNF-alpha) with its downstream signaling machinery promoting inflammation and cell death, were found in acute-on chronic ischemia. In chronic critical ischemia, gene expression changes were much less striking than in acute-on-chronic ischemia, with 74 genes significantly upregulated and 34 genes downregulated (change in 1.3% of all genes). In the chronic situation, the anabolic and survival factors, insulin-like growth factor 1 (IGF-1) and IGF-2, were upregulated in atrophic and regenerating myocytes together with attenuated HIF, VEGF, and VEGFR-2 expression in the same cells. In conclusion, acute-on-chronic and chronic human skeletal muscle ischemia result in distinct gene expression patterns. These findings may be of importance in the design of novel therapies, such as therapeutic vascular growth, for patients suffering from lower limb ischemia. PMID- 15135260 TI - Angiotensin-converting enzyme gene polymorphism and common carotid stiffness. The Rotterdam study. AB - The insertion/deletion (I/D) polymorphism of the ACE gene may be involved in structural arterial changes. Aim of the present study was to assess the relationship between the ACE I/D gene and vessel wall stiffness among older adults. The study was conducted within the Rotterdam study, a population-based cohort study including subjects aged 55 years and older. The II, ID and DD genotypes of the ACE gene were determined in all subjects. The distensibility coefficient (10(-3)/kPa) of the carotid artery and the carotid-femoral pulse wave velocity were measured during the third phase of the Rotterdam study (1997-1999) and were used as measure of arterial stiffness. Data on both carotid stiffness and the ACE genotype were available for 3001 participants. After adjustment for age and gender, subjects with the ID and DD genotype had higher carotid stiffness compared to subjects with II genotype (distensibility coefficient (10(-3)/kPa) 10.24 (95% CI, 10.06-10.43), 10.27 (95% CI, 10.02-10.52), 10.65 (95% CI, 10.37 10.93), respectively (ID versus II genotype, P = 0.017), (DD versus II genotype, P = 0.037)). In stratified analyses, the association was strongest in subjects younger than 70 years. No difference was seen for pulse wave velocity among genotypes. In conclusion, the results of this population-based study show that the ACE ID/DD genotypes are associated with higher common carotid stiffness. PMID- 15135261 TI - The effect of diet enriched with alpha-linolenic acid on soluble cellular adhesion molecules in dyslipidaemic patients. AB - BACKGROUND: Leukocyte adhesion and transendothelial migration, the critical pathogenic components in the development of atherosclerotic lesions, are largely mediated by cellular adhesion molecules (CAMs). We examined whether dietary supplementation with alpha-linolenic acid (ALA, 18:3n-3) affects the levels of soluble forms of CAMs in dyslipidaemic patients. METHODS: We recruited 90 male dyslipidaemic patients (mean age=51+/-8 years) following a typical Greek diet. They were randomly assigned either to 15 ml of linseed oil (rich in ALA) per day (n=60) or to 15 ml of safflower oil (rich in linoleic acid [LA, 18:2n-6]) per day (n=30). The ratio of n-6:n-3 in linseed oil supplemented group was 1.3:1 and in safflower oil supplemented group 13.2:1. Dietary intervention lasted for 12 weeks. Blood lipids, soluble intercellular adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1) and soluble E-selectin (sE-selectin) were measured. RESULTS: Dietary supplementation with ALA significantly decreased sVCAM-1 levels (median decrease 18.7% [577.5 ng/ml versus 487 ng/ml, P=0.0001]). In the LA supplemented group, sVCAM-1 was also significantly decreased but to a lesser extent (median decrease 10.6% [550.5 ng/ml versus 496 ng/ml, P=0.0001]). After controlling for smoking habits, no significant difference was observed in the reduction of sVCAM-1 levels between the two treatment arms (P=0.205). The decrease of sVCAM-1 was independent of lipid changes in both groups. CONCLUSIONS: Dietary supplementation with ALA for 12 weeks significantly decreases sVCAM-1 levels in dyslipidaemic patients. This effect presents a potential mechanism for the beneficial effect of plant n-3 polyunsaturated fatty acids in the prevention of coronary artery disease. In addition, dietary supplementation with LA significantly decreases sVCAM-1 levels, an effect which requires further investigation. PMID- 15135262 TI - Impact of chronic LDL-apheresis treatment on Achilles tendon affection in patients with severe familial hypercholesterolemia: a clinical and ultrasonographic 3-year follow-up study. AB - OBJECTIVE: Pain of Achilles tendon (AT) is a common symptom in patients with severe familial hypercholesterolemia (FH) and often associated with AT xanthomas. It is unknown if these changes are potentially reversible during lipid lowering treatment. LDL-apheresis (LA) represents the most effective lipid lowering regimen. Aim of this investigation was to determine clinical and ultrasonographic reduction of AT xanthomas in patients with severe FH undergoing regular LA. METHODS: At baseline, patient history of 22 patients with FH undergoing LA was obtained and their 44 ATs were evaluated clinically and with ultrasound for the presence of xanthomas. Three years later, both examinations of ATs could be repeated at follow-up visits in 16 patients. AT thickness and changes in echo structure were assessed at both points of time and compared to each other as well as to a healthy control group (n = 21). ROC analysis was performed to identify the optimal cut-off in AT thickness between healthy and affected ATs. RESULTS: Twelve of 22 FH patients suffered from AT pain at least once during their life time. At baseline, AT thickness was significantly increased compared to the healthy control group (mean sagittal diameter 10.1 +/- 3.6 mm). At follow-up, AT thickness was significantly reduced to 8.2 +/- 3.3 mm (mean) under LA, whereas changes in echo structure were less distinct between both visits. Conversely, in the control group, mean AT thickness was 5.2 +/- 0.6 mm. The optimal cut-off between healthy and affected ATs was determined to be 6mm. CONCLUSIONS: Ultrasonographic changes of the AT (thickening and changes in echo structure) are frequent in patients with severe FH, even if xanthomas are not clinically evident. LA treatment has the capability to reduce AT xanthomas and thickness. Ultrasound may give information about diagnosis and follow-up of AT affection in patients with FH. PMID- 15135263 TI - The effect of high dose atorvastatin therapy on lipids and lipoprotein subfractions in overweight patients with type 2 diabetes. AB - Few data are available on the effects of high dose statin therapy on lipoprotein subfractions in type 2 diabetes. In a double blind randomised placebo-controlled trial we have studied the effects of 80 mg atorvastatin over 8 weeks on LDL, VLDL and HDL subfractions in 40 overweight type 2 diabetes patients. VLDL and LDL subfractions were prepared by density gradient ultracentrifugation. Triglycerides, cholesterol, total protein and phospholipids were measured and mass of subfractions calculated. HDL subfractions were prepared by precipitation. Atorvastatin 80 mg produced significant falls in LDL subfractions (LDL(1) 66.2 mg/dl:36.6 mg/dl, LDL(2) 118:56.6 mg/dl, LDL(3) 36.9:19.9 mg/dl all P < 0.01 relative to placebo) and VLDL subfractions (VLDL(1) 55:22.1 mg/dl, VLDL(2) 40.1:19.1 mg/dl, VLDL(3) 52.6:30 mg/dl all P < 0.01 relative to placebo). There was no change in the proportion of LDL present as LDL(3). There was a reduction in the proportion of VLDL as VLDL(1) and a reciprocal increase in the proportion as VLDL(3). Changes in VLDL subfractions were associated with changes in lipid composition, particularly a reduction in cholesterol ester and a reduction in the cholesterol ester/triglyceride ratio. Effects on HDL subfractions were largely neutral. High dose atorvastatin produces favourable effects on lipoprotein subfractions in type 2 diabetes which may enhance antiatherogenic potential. PMID- 15135264 TI - Long-term reduction of C-reactive protein concentration by regular LDL apheresis. AB - BACKGROUND: Increased C-reactive protein (CRP) concentration is an independent risk predictor for coronary heart disease (CHD). A therapeutic reduction of CRP might, therefore, reduce the risk of cardiovascular events. A single LDL apheresis lowers LDL cholesterol by 45-70%, and CRP by 20-65%, however, less is known about the long-term effects of LDL apheresis on CRP levels. METHODS: We investigated 34 CHD patients (20 males, 14 females, 52 +/- 10 years) on statin therapy who were regularly treated by LDL apheresis because of drug-resistant hypercholesterolemia. Measurements of CRP (ultrasensitive nephelometric assay) were performed before the first apheresis, before a current apheresis (1 or 2 weeks after the last apheresis), and after a current apheresis (treatment period: 5.3 +/- 4.2 years, range: 0.25-12.5 years). LDL apheresis was performed by immunoadsorption (n = 6), dextran sulfate adsorption (n = 13), heparin-induced extracorporal LDL precipitation (HELP, n = 9), or direct adsorption of lipoproteins (DALI, n = 6). RESULTS: CRP was significantly lower before a current apheresis (median: 0.75 mg/l, range: 0.16-4.33 mg/l) compared to before the first apheresis (median: 0.85 mg/l, range: 0.16-7.02 mg/l; P < 0.05). As expected, total and LDL cholesterol were lower before a current apheresis compared to before the initial apheresis while fibrinogen concentration did not differ significantly. CONCLUSION: Over the period of more than 5 years LDL apheresis slightly, but significantly reduced CRP concentrations in patients with CHD on statin therapy, which may contribute to the stabilization of atherosclerosis in hypercholesterolemic patients treated with LDL apheresis. These results are even more impressive when the known age-related increase in CRP over the treatment period is taken into account. PMID- 15135265 TI - Prognostic value of plasma interleukin-6 concentrations and the -174 G > C and 572 G > C promoter polymorphisms of the interleukin-6 gene in patients with acute myocardial infarction treated with thrombolysis. AB - The present study was performed to investigate the prognostic value of plasma interleukin-6 (IL-6) concentrations and promoter polymorphisms of the IL-6 gene in patients with acute myocardial infarction treated with thrombolysis. Two hundred and eight patients with myocardial infarction treated with thrombolysis were included and followed for 2-5 years. Plasma concentrations of IL-6 were measured at admission and 48 h after admission. Genotyping for the -174 G > C and -572 G > C IL-6 polymorphisms was performed. Patients who died of cardiovascular causes or suffered a new myocardial infarction during follow-up had increased plasma concentrations of IL-6 at admission (P < 0.002) and at 48 h after admission (P < 0.05) compared with patients who had an uneventful course. IL-6 levels above the median at admission were independently associated with a worse prognosis. No associations were found between IL-6 levels and the promoter polymorphisms. The -174 G > C polymorphism was not associated with cardiovascular death or a new myocardial infarction, whereas the -572 G > C polymorphism showed a borderline significant increase in risk (P = 0.05) in univariate analysis. In conclusion, the early IL-6 response during myocardial infarction is associated with prognosis in patients with Q-wave myocardial infarction, whereas no associations were found between IL-6 genotype and phenotype. PMID- 15135266 TI - Autosomal recessive hypercholesterolaemia: long-term follow up and response to treatment. AB - Autosomal recessive hypercholesterolaemia (ARH) is caused by mutations in ARH on chromosome 1p35-36, encoding a putative adaptor protein. Mutations in the gene prevent normal internalisation of the low density lipoprotein (LDL) receptor by cultured lymphocytes and monocyte-derived macrophages, but not skin fibroblasts. This newly identified disorder is characterised by severe hypercholesterolaemia, large tendon, tuberous and planar xanthomas and premature atherosclerosis. We describe long-term (9-23 years) follow up and response to treatment of eight subjects with ARH from four families (Turkish/Lebanese, Indian-Asian, English and Italian). The clinical phenotype of ARH is similar to that of classical homozygous familial hypercholesterolaemia (FH) caused by mutations in the LDL receptor gene but is more variable, less severe and is more responsive to lipid lowering therapy with bile acid sequestrants and/or HMG-CoA reductase inhibitors. The latter reduced total serum cholesterol by up to 60% and the former by 20-35%. The cardiovascular complications of premature atherosclerosis seem to be delayed in some individuals and the involvement of the aortic root and valve are rarer in comparison with homozygous FH. PMID- 15135267 TI - Effects of oral and transdermal 17 beta-estradiol combined with progesterone on homocysteine metabolism in postmenopausal women: a randomised placebo-controlled trial. AB - Mild hyperhomocysteinemia is a risk factor for both ischaemic heart disease and venous thromboembolism. The effects of transdermal estrogen replacement therapy (ERT) on homocysteine metabolism in postmenopausal women have scarcely been investigated. This clinical trial aimed to estimate the effects of combined hormone replacement therapy on the fasting total homocysteine levels according to the estrogen route of administration. We enrolled 196 postmenopausal women, who were randomly allocated to receive on a continuous basis either 1mg of 17 beta estradiol orally (n = 63) or 50 microg transdermally (n = 68) per day, both combined with a daily intake of 100 mg progesterone, or placebo (n = 65) over a period of 6 months. Neither oral nor transdermal ERT significantly affected total plasma homocysteine levels or red-blood cell folate levels. However, oral ERT significantly decreased plasma vitamin B12 levels compared to placebo (mean relative variation difference over 6 months between oral ERT and placebo: -11.7% (95%CI, -21 to -2%) whereas transdermal ERT did not display any significant effects. Our data show that transdermal ERT as well as low dose of oral ERT does not significantly affect the homocysteine metabolism. This finding does not support a role for transdermal estrogen in the prevention of ischaemic heart disease in postmenopausal women. PMID- 15135268 TI - Genetic risk for restenosis after coronary balloon angioplasty. AB - Plain old balloon angioplasty (POBA) is a useful therapeutic strategy especially for angioplasty of small coronary arteries. An association study was performed to identify genes that confer susceptibility to restenosis after POBA. The study population comprised 730 individuals (424 men, 306 women) who underwent successful POBA in at least one major coronary artery and were examined angiographically 6 months after the procedure. A total of 469 subjects (273 men, 196 women) exhibited no restenosis after POBA for any of the coronary lesions, whereas 261 subjects (151 men, 110 women) manifested restenosis for all lesions. The genotypes for 40 polymorphisms of 34 genes were determined with a fluorescence- or colorimetry-based allele-specific DNA primer-probe assay. Multivariate logistic regression analysis with adjustment for age, body mass index, and the prevalence of smoking, hypertension, diabetes mellitus, hypercholesterolemia, and hyperuricemia revealed that two polymorphisms (242C --> T in the NADH/NADPH oxidase p22 phox (p22-PHOX) gene and 2136C --> T in the thrombomodulin (THBD) gene) in men and two polymorphisms (584G --> A in the paraoxonase 1 (PON1) gene and 2445G --> A in the fatty acid-binding protein 2 (FABP2) gene) in women were significantly associated with restenosis after POBA. A stepwise forward selection procedure revealed that the effects of these polymorphisms on restenosis were statistically independent of conventional risk factors for coronary artery disease. Genotyping of these polymorphisms may prove informative for assessment of genetic risk for restenosis after POBA. PMID- 15135269 TI - Increased soluble ICAM-1 and IL-6 are not speciofic plasma markers of endothelial dysfunction. PMID- 15135271 TI - Pravastatin in patients with nonalcoholic steatohepatitis: results of a pilot study. PMID- 15135272 TI - An evaluation of interface contact profiles in two low contact bone plates. AB - Bone plate design has evolved dramatically in recent years. The Dynamic Compression Plate (DCP) has been superseded by bi- and uni-cortical plates that claim a reduced interface contact between the plate and the underlying bone. It is believed that contact reduction ameliorates the localised ischaemia that develops subsequent to plate application. In this study, the interface characteristics of the Limited Contact-Dynamic Compression Plate (LC-DCP) and the Contour Plus (CP) plating systems have been quantitated using Fuji prescale pressure sensitive film interposed between the plate and the bone. Ten-hole plates were applied to the same aspect of either the humeral, radial or ulnar diaphysis of human cadaveric bone in a reproducible manner. The average pressure, force and interface contact area were calculated using Interactive Data Language (IDL) image analysis software. The CP system was consistently lower, in terms of interface contact, than the LC-DCP in each of the specimen locations tested (P<0.0001). The CP system displayed a 'point-contact' configuration along the interface with high pressures recorded at these points, the significance of which is unknown. PMID- 15135273 TI - Reliability of radiographs in defining union of internally fixed fractures. AB - We assessed whether radiographs can predict union of internally fixed fractures, and therefore be used as end-points in studies of fracture healing. Forty-seven radiographic series of forearm, femoral and tibial fractures treated by internal fixation over a 3-year period were reviewed. All forearm fractures were treated with dynamic compression plates (DCP), and all tibial and femoral fractures with intra-medullary nails. Callus formation and fracture line filling with time were measured on each radiograph. The ability of five orthopaedic surgeons to chronologically rank the blinded radiographs and to agree on the point of union was assessed. Correlation between callus formation, fracture line filling and union was noted. The ability of surgeons to correctly rank the radiographs and to agree on the point of union was in the order of 70%. Callus formation and union progression was significant in femoral fractures (P<0.05). Fracture line filling and union progression showed significance in the forearm (P<0.01) and femoral groups (P<0.05). Taking serial radiographs to assess healing would have led to only one early intervention. Radiographs do not define union in internally fixed fractures with sufficient accuracy to enable their use as end-points of fracture healing. Studies quoting radiographic end-points should be interpreted with care. PMID- 15135274 TI - Is ultrasound really helpful in the detection of rib fractures? AB - OBJECTIVE: To determine the usefulness of ultrasound in the detection of rib fractures. PATIENTS AND METHODS: A prospective study was performed over a 3-month period. Patients presenting with a high clinical suspicion of rib fracture(s) to the Accident and Emergency Department were referred for radiological work-up with a PA chest radiograph, an oblique rib view and a chest ultrasound. Associated lesions, e.g. pleural effusion, splenic laceration and pneumothorax were recorded. RESULTS: Fourteen patients were radiologically assessed. The mean patient age was 31 years (range 16-55 years) and the M:F ratio 3.7:1 (11 men and 3 women). Ten patients displayed a total of 15 broken ribs. Chest radiography detected 11, oblique rib views 13 and ultrasound 14 broken ribs. Ultrasound findings included discontinuity of cortical alignment in 12 fractures, an acoustic linear edge shadow in nine and a reverberation artifact in six. Concordance with plain film findings, and especially oblique rib views, was good, though better when the rib fractures fragments were markedly displaced. One splenic laceration was detected with an associated small pleural effusion. There were no pneumothoraces. The average time of ultrasound examination was 13 min. CONCLUSION: Ultrasound does not significantly increase the detection rate of rib fractures, may be uncomfortable for the patient and is too time-consuming to justify its routine use to detect rib fractures. PMID- 15135275 TI - Fluoroscopic guidance versus surgical navigation for distal locking of intramedullary implants. A prospective, controlled clinical study. AB - A prospective controlled clinical study was performed to compare fluoroscopic guidance with fluoroscopy-based surgical navigation for distal locking of intramedullary implants. Forty-two patients with fractures of the lower extremity treated by intramedullary nailing were divided in two groups: distal locking either with fluoroscopic guidance (group I) or with surgical navigation (group II). The average fluoroscopic time to insert one interlocking screw with fluoroscopic guidance was 108 s compared with 7.3s in the navigation group. The average procedure time to insert one interlocking screw in group I was 13.7 min compared with 17.9 min in group II. The drill bit failed to pass through the interlocking hole in one patient from group II. There was no significant difference in the technical reliability between both groups. Fluoroscopic times to achieve equivalent precision are reduced with fluoroscopy-based surgical navigation compared with fluoroscopic guidance. Fluoroscopy-based surgical navigation is recommended for intraoperative guidance in situations where reduction of exposure to radiation is considered advantageous over the increase of procedure time. PMID- 15135276 TI - Long-term functional outcome assessment of plate fixation and autogenous bone grafting for clavicular non-union. AB - INTRODUCTION: The aim of our study was to assess the long-term functional and radiological outcome of clavicular non-union in patients treated with open reduction and bone grafting. A total of 24 non-unions treated between 1994 and 2001 were analysed using chart and radiological review and assessed with the American Academy of Orthopaedic Surgeons (AAOS) DASH questionnaire. RESULTS: There were 13 males and 10 females with a mean age of 37.95 (range 21-65) years. One patient had bilateral injuries. The average time from injury to operation was 10.3 (range 4-29) months and the average follow-up post-operatively was 42.1 (range 6-75) months. All patients were treated using a dynamic compression or reconstruction plate with autogenous bone grafting. Twenty-two of the 24 non unions eventually healed. DASH assessment indicated a higher level of disability in the treated group than found in the normal population. This only proved significant in the sub-population of patients with associated co-morbidities. CONCLUSION: We conclude that the long-term outcome results of this procedure indicate it to be a well-tolerated and successful operation in treating the disability and pain associated with clavicular non-union. Most patients return to a daily level of function close to the general population. Patients with associated co-morbidities should have these treated pre-operatively for best results. PMID- 15135277 TI - Plating of humeral shaft fractures--has the pendulum swung back? AB - We reviewed 49 patients following plate osteosynthesis of humeral shaft fractures. There were no complications as a result of surgery. Union occurred in 47 patients (96%) at a mean of 9 weeks. Two patients required secondary procedures to achieve union. All patients had full range of motion in the elbow and shoulder joints following union. In the light of the popularisation of intramedullary nailing techniques in the last decade, with recognised complications of iatrogenic radial nerve injury, inadequate rotational stabilisation, non-union and shoulder impingement, we advocate plating of humeral shaft fractures as the surgical treatment of choice. PMID- 15135278 TI - Bridging plate osteosynthesis of humeral shaft fractures. AB - This study was approved by the Ethics Committee of the Faculty of Medical Sciences and developed during November 2000 and July 2001 in the Orthopedic and Traumatology Department of UNICAMP. There were 15 patients, 11 males, age between 14 and 66 years. All fractures were unilateral. Of the 15 patients eight were polytraumatised, two of them had open fractures. The others had an isolated fracture of the humerus, of which one was open. None of the patients had previous lesions of the radial nerve, but in two patients there was a lesion of the brachial plexus. All of the patients underwent a bridging plate osteosynthesis of the humeral shaft fractures using only two small incisions proximal and distal to the fracture site. We used broad or narrow D.C.P. plates for large fragments mostly with 12 holes, fixed with two or three screws at each end. All cases united with an average time of 8-12 weeks, with the exception of one case with a grade III open fracture and a brachial plexus lesion on the same side. We had no major complications. All patients recovered good function of the limb without significant residual deformity. PMID- 15135279 TI - Carbon fibre plates in the treatment of femoral periprosthetic fractures. AB - A retrospective review of fractures occurring around the femoral component of total hip replacements was performed using patient notes and X rays. Between 1996 and 2002, 12 patients with total hip replacement had fixation of their periprosthetic femoral shaft fracture with carbon fibre plates. The mean patient age was 76 years (range 57-94). With regard to fracture type (Johansson Classification), three were Type 2 and nine were Type 3. In 11 cases the average time to union was 4 months (range 3-6). There was one non-union leading to implant failure. This was subsequently treated with a revision arthroplasty. There were no other significant complications. Although designed to fit the supracondylar region of the femur, we had no difficulty in applying the plate over the trochanteric region. Our results indicate that carbon fibre plates are effective in the management of femoral periprosthetic fractures in the elderly and this may be related to the unique properties of the plate. PMID- 15135280 TI - Distal femoral fractures: long-term outcome following stabilisation with the LISS. AB - We studied 29 patients with distal femoral fractures stabilised using the less invasive stabilisation system (LISS). Four patients were excluded from the final follow-up (three deaths and one case of quadriplegia). The mean age of the remaining 25 patients (9 males and 16 females) was 60.9 years and the mean follow up 18 months (range 12-24 months). Eleven patients were tertiary referrals from other hospitals (seven cases were referred due to failure of primary fixation). Overall, there were 12 cases of high-energy trauma (7 open fractures). According to the AO classification, there were 5 Type 33A, 2 Type 33B and 12 Type 33C fractures and 4 Type 32A, 1 Type 32B, 1 Type 32C fractures. Functional assessment was performed using the modified Hospital for Special Surgery (HSS) and the Schatzker and Lambert scores. The average time to union in 22 cases was 3.5 months (range 2-5 months). All of the acute cases united without the need for bone grafting. There were three out of seven cases of non-union in the salvage group still undergoing treatment. The overall result in the acute cases was good and in the salvage cases fair. While this is a small series of patients, our preliminary data indicate favourable results using the LISS in stabilising acute distal femoral fractures. However, when the LISS is used as a revision tool the results seem to be less satisfactory. The system appears to be user-friendly and no technical difficulties were encountered. PMID- 15135281 TI - Percutaneous plating of distal tibial fractures. Preliminary results in 21 patients. AB - We describe the surgical technique and report the outcome after closed reduction and percutaneous plating in 21 closed extraarticular distal tibial fractures using titanium LC-DCP. A long plate was pushed subcutaneously through a small incision at the medial malleolus and fixed with screws through stab incisions. Fracture reduction was anatomical or nearly anatomical without angular displacement in 14 cases, and considered acceptable in four cases. Two patients were reoperated because of malreduction. Seventeen fractures healed within 6 months. There were two delayed unions, and two non-unions. There were two deep infections, both in diabetic patients. Follow-up by an independent observer at on average 14 (5-25) months showed various symptoms such as slightly reduced ankle mobility (9/20), reduced walking ability (11/20) or tenderness around the plate (11/20). No patient complained of knee symptoms related to the surgery. Operative treatment of closed distal tibial shaft fractures with a long titanium plate provided good anatomical results and allowed in most patients early weight bearing. In patients with soft tissues in good condition and no risk factor for infection percutaneous plate osteosynthesis might become an attractive treatment option for fractures in the distal tibia. PMID- 15135282 TI - Fractures of the distal tibia: minimally invasive plate osteosynthesis. AB - Unstable fractures of the distal tibia that are not suitable for intramedullary nailing are commonly treated by open reduction and internal fixation and/or external fixation, or treated non-operatively. Treatment of these injuries using minimally invasive plate osteosynthesis (MIPO) techniques may minimise soft tissue injury and damage to the vascular integrity of the fracture fragments. We report the results of 20 patients treated by MIPO for closed fractures of the distal tibia. Their mean age was 38.3 years (range: 17-71 years). Fractures were classified according to the AO system, and intra-articular extensions according to Ruedi and Allgower. The mean time to full weight-bearing was 12 weeks (range: 8-20 weeks) and to union was 23 weeks (range: 18-29 weeks), without need for further surgery. There was one malunion, no deep infections and no failures of fixation. MIPO is an effective treatment for closed, unstable fractures of the distal tibia, avoiding the complications associated with more traditional methods of internal fixation and/or external fixation. PMID- 15135283 TI - What do they do with their metalwork? What patients do with orthopaedic implants given to them after surgery. AB - Removal of metalwork is one of the commonest orthopaedic elective procedures with patients often requesting to keep their implants. Five hundred consecutive patients who had removal of their orthopaedic implant between April 1994 and March 2000 were contacted by post. Three hundred and seventy-two responded (response rate 74%). Two hundred and thirty-nine patients (64%) had metalwork extracted from their lower limb, whilst one hundred and thirty-three (36%) had metalwork extracted from their upper limb. Sixty-one patients (17%) had kept their metalwork upon being contacted. Of the sixty-one patients, 10 (16%) had used their implant for functional use, 22 (36%) had stored theirs and 29 (48%) did not know what had happened to theirs. PMID- 15135284 TI - Technical difficulty of metal removal after LISS plating. PMID- 15135285 TI - Capture of the screw during percutaneous screw insertion. AB - The simple technique to maintain the engagement of the screw and the screw driver during percutaneous locking screw insertion is presented. The method can capture the screw when the screw becomes disengaged. This will be helpful when percutaneous screw insertion is performed in the narrow and deep location of the bone such as a locking screw at the upper part of the femur during retrograde femoral nailing. PMID- 15135286 TI - Minimal invasive percutaneous plate osteosynthesis for complex monteggia fracture with type III coronoid process fracture. PMID- 15135287 TI - Sternoclavicular hyperostosis with pathological fracture of the clavicle--a case report. PMID- 15135289 TI - A fail safe method of digital tourniquet. PMID- 15135290 TI - Whither T-suppressors: if they didn't exist would we have to invent them? AB - Arriving at an understanding of the role of suppressor T-cells (regulatory T cells, CD4(+)CD25+) depends on whether their functional repertoire is somatically selected to be anti-Self or anti-Nonself. Immunologists are ambivalent; often publications espousing opposite views share an author. Here the arguments are detailed that the suppressor repertoire is not somatically selected to be anti Self, but rather it is anti-Nonself. Therefore, suppression cannot regulate the Self-Nonself discrimination; its function is to regulate the magnitude and class of the anti-Nonself effector response. PMID- 15135291 TI - Reduced L-selectin (CD62LLow) expression identifies tumor-specific type 1 T cells from lymph nodes draining an autologous tumor cell vaccine. AB - Reduced expression of CD62L can identify tumor-specific T cells in lymph nodes draining murine tumors. Here, we examined whether this strategy could isolate tumor-specific T cells from vaccinated patients. Tumor vaccine-draining lymph node (TVDLN) T cells of seven patients were separated into populations with reduced (CD62LLow) or high levels of CD62L (CD62LHigh). Effector T cells generated from CD62LLow cells maintained or enriched the autologous tumor specific type 1 cytokine response compared to unseparated TVDLN T cells in four of four patients showing tumor-specific cytokine secretion. Interestingly, effector T cells generated from CD62LLow or CD62LHigh TVDLN were polarized towards a dominant type 1 or type 2 cytokine profile, respectively. For CD62LLow T cells the type 1 cytokine profile appeared determined prior to culture. Since a tumor-specific type 1 cytokine profile appears critical for mediating anti-tumor activity in vivo, this approach might be used to isolate T cells for adoptive immunotherapy. PMID- 15135292 TI - Induction of dendritic cell maturation by IL-18. AB - IL-18 is a pluripotent proinflammatory cytokine produced primarily by antigen presenting cells involved in numerous aspects of immune regulation most notably on lymphoid cells. The effect of IL-18 stimulation on cells in the myeloid compartment, however, has been poorly studied. Human monocytes did not respond to IL-18. However, the human myelomonocytic cell line KG-1 and monocyte-derived dendritic cells (generated by GM-CSF+IL-4) showed a marked increase in CD83, HLA DR, and several costimulatory molecules upon stimulation with IL-18. Furthermore, IL-18 decreased pinocytosis of these cells and increased their ability to stimulate alloreactive T cell proliferation, all characteristics of mature dendritic cells. These results suggest that IL-18 is involved in the maturation of myeloid DCs, but not differentiation of monocytes into DCs. The finding that IL-18 is involved in the maturation of dendritic cells is both novel and unexpected and indicates another important role for IL-18 as a key regulator of immune responses. PMID- 15135293 TI - The role of CD1d in the immune response against Listeria infection. AB - To address the role of CD1d in mucosal immune regulation in bacterial infection, we infected CD1d KO mice with Listeria monocytogenes (Lm). A higher systemic bacterial burden associated with inflammatory lymphocytic infiltrations within the intestine was found in CD1d KO compared with wild type (WT) mice. Lm induced strong IFN-gamma mRNA expression in the liver of WT and the intestine of CD1d KO mice, thus demonstrating the dual, opposing immune activities of IFN-gamma in Lm infection that is dependent on CD1d and/or NKT cells. Analysis of hepatic T cell population demonstrated a reduction of NK1.1(+)TCRbeta+ cells in both mice, followed by recovery only in WT mice. Last, the proportion of alpha4beta1 integrin on lung lymphocytes from CD1d KO was dramatically increased compared with WT mice. Thus, the absence of CD1d resulted in increased susceptibility towards Listeria infection, induced changes in NKT cells, and increased trafficking of alpha4beta1 molecule to inflamed lung. PMID- 15135294 TI - Differential effect of IL-4 and IL-13 on the expression of recombination activating genes in mature B cells from human peripheral blood. AB - We examined the expression of recombination-activating genes (RAG-1 and RAG-2) and activation-induced cytidine deaminase (AID) by mature human blood B cells stimulated with anti-CD40 in the presence of IL-4 or IL-13. IL-4 was an effective cofactor for RAG-1 and RAG-2 expression, whereas IL-13 was not. In addition, IL-4 dependent RAG expression combined with AID and IgE expression allowed predominant expression of newly rearranged lambda light chains on IgE+ cells generated from kappa+ cells. Although the magnitudes of IL-4- and IL-13-dependent AID and IgE expression were related to expression levels of binding subunits of the IL-4 and IL-13 receptors, IL-13 was ineffective for light chain replacement in the induced IgE+ cells due to the failure in RAG expression. Our studies using mature blood B cells indicate that IL-4-responsive cells, unlike IL-13-responsive cells, undergo lambda gene rearrangement leading to replacement in parallel with RAG expression and suggest that this replacement may contribute to the regulation of affinity maturation of IgE antibodies. PMID- 15135295 TI - The T cell activation marker CD150 can be used to identify alloantigen-activated CD4(+)25+ regulatory T cells. AB - We have been investigating whether alloantigen-specific CD4(+)25+ regulatory T cells can be identified for use in treating graft-versus-host disease. CD150, which is upregulated on the surface of all activated T lymphocytes, was identified as a candidate marker for alloantigen-activated CD4(+)25+ regulatory T cells by gene chip analysis. Freshly isolated CD4(+)25+ cells had only low cell surface expression of CD150, comparable to that of CD4(+)25- T cells. Increased CD150 expression was observed on all T cells after coculture with allogeneic stimulator cells. When purified CD4(+)25+ cells were precultured with allogeneic stimulator cells, then sorted into CD150+ and CD150- subsets, allosuppressive activity was contained primarily in the CD150+ fraction. These cells also suppressed the proliferation of alloantigen-activated autologous T cells, and they could be expanded in vitro without loss of their suppressive capacity. These results suggest that CD150 can be used as a marker for the identification of purified alloantigen-activated CD4(+)25+ regulatory T cells. PMID- 15135296 TI - Implications of proteasome inhibition: an enhanced macrophage phenotype. AB - The objective of this study was to elucidate the role of the cellular proteasome on endotoxin-mediated activation of the macrophage. To study this role, THP-1 cells were stimulated with lipopolysaccharide (LPS) with selective cells being pretreated with the proteasome inhibitor, lactacystin or MG-132. LPS stimulation led to the phosphorylation and degradation of IRAK, followed by activation of JNK/SAPK, ERK 1/2, and p38. Subsequently, LPS induced the degradation of IkappaB, and the nuclear activation of NF-kappaB and AP-1. Activation of these pathways was associated with the production of IL-6, IL-8, IL-10, and TNF-alpha. Proteasome inhibition with either lactacystin or MG-132 attenuated LPS-induced IRAK degradation, and enhanced activation of JNK/SAPK, ERK 1/2, and p38. Proteasome inhibition, also, led to increased LPS-induced AP-1 activation, and attenuated LPS-induced IkappaB degradation resulting in abolished NF-kappaB activation. Proteasome inhibition led to significant modulation of LPS-induced cytokine production; increased IL-10, no change in IL-6, and decreased IL-8, and TNF-alpha. Thus, this study demonstrates that cellular proteasome is critical to regulation of LPS-induced signaling within the macrophage, and inhibition of the proteasome results in a conversion to an anti-inflammatory phenotype. PMID- 15135297 TI - High avidity CD8+ T cells generated from CD28-deficient or wildtype mice exhibit a differential dependence on lipid raft integrity for activation. AB - CD28 has been shown to play an important role in T cell activation. Among the downstream events associated with CD28 engagement is the reorganization of the cytoskeleton resulting in lipid raft aggregation. In our previous studies we investigated the involvement of lipid rafts in the activation of high avidity CD8+ T lymphocytes, which recognize cells bearing very low levels of peptide antigen, versus low avidity cells, which require high levels of peptide antigen. In these studies we found that high avidity cells were much more sensitive to lipid raft disruption compared to low avidity cells. Given the important role for CD28 in lipid raft reorganization and our previous finding that high avidity cells are extremely dependent on lipid raft integrity, we hypothesized that high avidity cells could not be generated in the absence of CD28. Surprisingly, we have found that the absence of CD28 does not alter the ability to generate high or low avidity CD8+ T cells. In fact high and low avidity lines generated in parallel from CD28-deficient and WT mice exhibited very similar requirements for peptide antigen. We next compared the effect of lipid raft disruption on the activation of high versus low avidity cells from CD28-deficient and WT mice. While high avidity cells generated from WT mice exhibited the expected dependence on lipid raft integrity, high avidity cells from CD28-deficient mice were not affected. These data suggest that the lines generated from the CD28-deficient mice have developed alternative strategies to promote high sensitivity to peptide antigen. PMID- 15135298 TI - Activation of latent transforming growth factor beta 1 and inhibition of matrix metalloprotease activity by a thrombospondin-like tripeptide linked to elaidic acid. AB - Impaired wound healing and skin aging are characterized by neutral protease mediated destruction of matrix macromolecules associated with disturbance in tissue repair. We synthesized a fatty acyl-peptide derivative at aims to simultaneously activate latent TGF-beta through its peptide domain, KFK, and inhibit MMPs through its lipophilic moiety, elaidic acid. Elaidyl-KFK as well as KFK were shown to activate LAP-TGF-beta both in vitro, using a solid phase assay with immobilized LAP-TGF-beta, and ex vivo using human dermal fibroblasts cultures. In both assays, as much as up to 10% of LAP-TGF-beta added could be recovered as active form. KQK, KQFK as well as their lipopeptide counterparts were inactive. Elaidyl-KFK-mediated LAP-TGF-beta activation led to up-regulation of collagen and TIMP-1 production and down regulation of PMA-induced MMP-1 expression in fibroblasts cultures. Those effects could be suppressed by supplementing cell culture medium with blocking TGF-beta antibody. Elaidyl-KFK inhibited MMP-2, MMP-9, MMP-3, MMP-1, in vitro with IC(50) equal to 1.2, 1.0, 0.24 and 8.9 microM, respectively. Its ex vivo inhibitory capacity, as assessed using skin tissue sections, towards the elastin-degrading capacity of MMP-9 was even more pronounced. At a 1 microM concentration, the lipopeptide decreased by up to 80% enzyme activity. Thus, "Lipospondin," i.e. elaidyl-KFK might be considered as a promising model compound to prevent age-associated dermal alterations. PMID- 15135299 TI - Melatonin-mediated regulation of human MT(1) melatonin receptors expressed in mammalian cells. AB - In mammals, the pineal hormone melatonin activates G protein-coupled MT(1) and MT(2) melatonin receptors. Acute exposure of recombinant MT(1) and MT(2) melatonin receptors to supraphysiological concentrations of melatonin differentially regulates these two receptors with the MT(2), but not the MT(1), exhibiting rapid desensitization and internalization. In the present study, we sought to determine whether prolonged exposure to supraphysiological and physiological concentrations of melatonin desensitized and/or internalized the MT(1) melatonin receptor. Using a Chinese hamster ovary (CHO) cell line stably expressing MT(1)-FLAG or transiently expressing MT(1)-green fluorescent protein (GFP) melatonin receptors, we found that prolonged exposure (8h) to supraphysiological concentrations of melatonin (100 nM) significantly increased the number of MT(1) melatonin receptors and decreased the affinity (K(i)) of melatonin for competition for 2-[125]iodomelatonin. A similar treatment also desensitized the MT(1) melatonin receptor-mediated stimulation of [(35)S]GTPgammaS binding, but did not internalize the receptor. In contrast, prolonged exposure to a concentration of melatonin mimicking nocturnal levels (400 pM) did not affect the number of MT(1) melatonin receptors, the affinity for melatonin, or the functional sensitivity of the receptor. We conclude that in vivo endogenous melatonin does not significantly affect the functional sensitivity of MT(1) melatonin receptors, however, exogenous melatonin taken therapeutically at doses above physiological levels could desensitize the receptor thereby affecting physiological responses mediated following activation of MT(1) melatonin receptors. PMID- 15135300 TI - Genistein inversely affects tubulin-binding agent-induced apoptosis in human breast cancer cells. AB - Genistein, a natural isoflavone phytoestrogen present in soybeans, has been extensively studied as a chemopreventive or therapeutic agent in several types of cancer. The traditional Asian diet is rich in soy products may explain in part why the incidence of breast cancer in Asian women is relatively low. To improve therapeutic benefits, we investigated the combination of genistein with chemotherapeutic agents in phenotypically dissimilar human breast cancer cells, MCF-7 and MDA-MB-231, in which estrogen receptor expression is positive and negative, respectively. In the present study, genistein significantly decreased cell apoptosis induced by tubulin-binding agents, paclitaxel and vincristine. FACScan analysis revealed that genistein also diminished the accumulation of the G2/M phase in the cell cycle caused by tubulin-binding agents. In situ staining of microtubules revealed that genistein could decrease paclitaxel-induced tubulin polymerization. However, in vivo tubulin polymerization assay revealed that simultaneous treatment of genistein did not change the tubulin/microtubule dynamic. Genistein reduced Bcl-2 phosphorylation triggered by paclitaxel and vincristine without changing Bax protein expression. p53 and p21 expression, monitored by Western blotting, was not altered by genistein. However, the expression of cyclin B1 and CDC2 kinase was markedly decreased in combination with genistein. In conclusion, genistein inversely affected tubulin-binding agent induced apoptosis via down-regulation of cyclin B1/CDC2 kinase expression resulting in reduced Bcl-2 phosphorylation. PMID- 15135301 TI - Inverse agonist properties of atypical antipsychotic drugs. AB - Mechanisms of action of several atypical antipsychotic drugs have been examined at the D(2) dopamine receptor expressed in CHO cells. The drugs tested were found to exhibit inverse agonist activity at the D(2) dopamine receptor based on their effects to potentiate forskolin-stimulated cyclic AMP (cAMP) accumulation. Each of the antipsychotic drugs tested (clozapine, olanzapine, quetiapine and risperidone) increased cAMP accumulation to the same extent. The increase in cAMP was also similar to that seen with typical antipsychotic drugs. Inverse agonism at the D(2) dopamine receptor seems, therefore, to be a property common to all classes of antipsychotic drugs. The effect of sodium ions on the binding of the drugs to the receptor was also assessed. Each of the atypical antipsychotic drugs tested here bound with higher affinity in the absence of sodium ions. Previous studies have shown that some antipsychotic drugs are insensitive to sodium ions and some bind with higher affinity in the presence of sodium ions. Given that all of these antipsychotic drugs are inverse agonists, it may be concluded that this sodium ion sensitivity is unrelated to mechanisms of inverse agonism. PMID- 15135302 TI - Induction of apoptosis in two human leukemia cell lines as well as differentiation in human promyelocytic cells by cyanidin-3-O-beta glucopyranoside. AB - Little is known about the potentially chemopreventive mechanisms of anthocyanins apart from their antioxidant activity. We investigated the in vitro capacity of the anthocyanin cyanidin-3-O-beta-glucopyranoside (Cy-g) to induce apoptosis in T lymphoblastoid, as well as apoptosis and differentiation in HL-60 promyelocytic cells. Although Cy-g-induced apoptosis (as well as necrosis) in the two systems, HL-60 cells were much less sensitive than T-lymphoblastoid cells. Moreover, treatment of HL-60 cells with Cy-g caused differentiation into macrophage-like cells and granulocytes. Concerning the mechanism of action, the induction of apoptosis in Jurkat T cells can be explained by a modulation of p53 and bax protein expression. At the molecular level, the induction of apoptosis and cytodifferentiation in HL-60 cells involved different proteins, thus suggesting that the effects of Cy-g on apoptosis and cytodifferentiation induction are two distinct events. These interesting biological properties should encourage further investigation into the chemopreventive and/or chemotherapeutic potential of Cy-g. PMID- 15135303 TI - Pharmacological characterization of a human-specific peroxisome proliferater activated receptor alpha (PPARalpha) agonist in dogs. AB - Peroxisome proliferator-activated receptor alpha (PPARalpha) is a key regulator in lipid metabolism and a potential therapeutic target for lipid-related metabolic diseases. It has been shown that there are species differences between human and mouse in response to several PPARalpha agonists in a transactivation assay. In the present study, we cloned a full length of dog PPARalpha and investigated the effects of a novel and potent agonist (KCL) for human PPARalpha. In a transactivation assay using the full length of PPARalpha, agonistic activity of KCL for dog PPARalpha (EC(50): 0.007 microM) was comparable to that for human PPARalpha (EC(50): 0.003 microM), but not that for rat PPARalpha (EC(50): 11.49 microM). Similar results were obtained from a transactivation assay using a GAL4/PPARalpha ligand-binding domain (LBD) chimera. A point-mutation study showed that I272 on PPARalphaLBD is a major contributor to species differences in response to KCL between human, dog, and rat PPARalpha. KCL also induced mRNA levels of HMG-CoA synthase in dog hepatocytes. When administered orally to dogs and rats, KCL significantly decreased plasma triglyceride levels in a dose dependent manner. The triglyceride-lowering effects of KCL in dogs were >100-fold more potent than those in rats. These results suggest that KCL may induce activation of highly potent PPARalpha in humans as well as dogs, and that dog is a suitable animal model for studying and predicting the biological actions of potent agonists for human PPARalpha. PMID- 15135304 TI - Selenium compounds modulate the calcium release channel/ryanodine receptor of rabbit skeletal muscle by oxidizing functional thiols. AB - Selenium compounds, such as sodium selenite and Ebselen were shown to increase high affinity ryanodine binding to the skeletal muscle type ryanodine receptor (RyR1) at nanomolar concentrations, and inhibit the receptor at low micromolar concentrations. This biphasic response was observed in both concentration and time-dependent assays. Extensive washing did not reverse either the stimulation or suppression of receptor binding, but both were prevented or reversed by addition of reduced glutathione, GSH. Selenium compounds were also shown to induce Ca(2+) release from the isolated sarcoplasmic reticulum vesicles. Sodium selenite and Ebselen stimulated the skeletal muscle ryanodine receptor by oxidizing 14 of 47 free thiols per monomer on RyR1 (as detected with the alkylating agent 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin) (CPM). Oxidation of the remaining thiols by these selenium compounds resulted in inhibition of the ryanodine receptor. PMID- 15135305 TI - Metformin (Glucophage) inhibits tyrosine phosphatase activity to stimulate the insulin receptor tyrosine kinase. AB - Metformin is a commonly used anti-diabetic but whether its mechanism involves action on the insulin receptor or on downstream events is still controversial. With a time course that was slow compared with insulin action, metformin increased tyrosine phosphorylation of the regulatory domain of the insulin receptor (specifically, tyrosine residues 1150 and 1151). In a direct action, therapeutic levels of metformin stimulated the tyrosine kinase activity of the soluble intracellular portion of the beta subunit of the human insulin receptor toward a substrate derived from the insulin receptor regulatory domain. However, metformin did not alter the order of substrate phosphorylation by the insulin receptor kinase. Using a Xenopus oocyte preparation, we simultaneously recorded tyrosine kinase and phosphatase activities that regulate the insulin receptor by measuring the tyrosine phosphorylation and dephosphorylation of peptides derived from the regulatory domain of the human insulin receptor. In an indirect stimulation of the insulin receptor, metformin inhibited endogenous tyrosine phosphatases and purified human protein tyrosine phosphatase 1B that dephosphorylate and inhibit the insulin receptor kinase. Thus, there was evidence that metformin acted directly upon the insulin receptor and indirectly through inhibition of tyrosine phosphatases. PMID- 15135306 TI - Quaternary ammonium-linked glucuronidation of tamoxifen by human liver microsomes and UDP-glucuronosyltransferase 1A4. AB - Tamoxifen (TAM), a nonsteroidal antiestrogen, is the most widely used drug for chemotherapy of hormone-dependent breast cancer in women. In the present study, we found a new potential metabolic pathway of TAM via N-linked glucuronic acid conjugation for excretion in humans. TAM N(+)-glucuronide was isolated from a reaction mixture consisting of TAM and human liver microsomes fortified with UDP glucuronic acid (UDPGA) and identified with a synthetic specimen by high performance liquid chromatography-electrospray ionization-mass spectrometry. However, no TAM-glucuronidating activity was detected in microsomes from rat, mouse, monkey, dog, and guinea pig livers. A strong correlation (r(2) =0.92 ) was observed between N-glucuronidating activities toward TAM and trifluoperazine, a probe substrate for human UDP-glucuronosyltransferase (UGT) 1A4, in human liver microsomes from eight donors (five females, three males). However, no correlation ( (r(2) =0.02 )) was observed in the activities between 7-hydroxy-4 (trifluoromethyl)coumarin and TAM. Only UGT1A4 catalyzed the N-linked glucuronidation of TAM among recombinant UGTs (UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A9, UGT2B4, UGT2B7, UGT2B15, and UGT2B17) expressed in insect cells. Apparent K(m) values for TAM N-glucuronidation by human liver microsomes and recombinant UGT1A4 were 35.8 and 32.4 microM, respectively. These results strongly suggested that UGT1A4 could play a role in metabolism and excretion of TAM without Phase I metabolism in human liver. TAM N(+)-glucuronide still had binding affinity similar to TAM itself for human estrogen receptors, ERalpha and ERbeta, suggesting that TAM N(+)-glucuronide might contribute to the biological activity of TAM in vivo. PMID- 15135307 TI - Transinactivation of the epidermal growth factor receptor tyrosine kinase and focal adhesion kinase phosphorylation by dietary flavonoids: effect on invasive potential of human carcinoma cells. AB - Focal adhesion kinase (FAK), a member of a growing family of structurally distinct protein tyrosine kinases (PTK), has been linked to specific phosphorylation events, and the elevation of FAK activity in human carcinoma cells correlated with increased invasive potential. Transactivation of the epidermal growth factor receptor (EGFR) tyrosine kinase activity is proposed to stimulate cell migration and the subsequent activation of downstream signaling pathways. Quercetin (Qu) and luteolin (Lu), are potent PTK inhibitors as well as putative chemopreventive agents. The present work, we demonstrate that Qu and Lu at concentration of 20 microM transinactivated EGFR tyrosine kinase activity with marked reduction in phosphotyrosyl level of 170, 125, 65, 60 and 42 kDa cellular proteins, and induced apoptosis in MiaPaCa-2 cells. The 125 kDa protein was further identified as a FAK by immunoprecipitation and immunoblotting analyses. Tumor cells treated with Lu or Qu dampened the phosphorylation of FAK. In addition, our data clearly demonstrated that tumor cells responded to Qu and Lu by parallel reductions in the levels of phosphorylated FAK and the secreted matrix metalloproteinase (MMP) that may lead to the suppression of invasive potential and cell migration in vitro. While the molecular mechanism of FAK regulation of MMP secretion in tumor cells remains unclear, our results suggested that blockade of the EGFR-signaling pathway may contributed to the net effect. As suggested in the current study, targeting EGFR and FAK with the objective of modulating their regulatory pathways could offer prospects for the treatment of EGFR-responsive cancers in the future. PMID- 15135308 TI - Pharmacological characterization of human excitatory amino acid transporters EAAT1, EAAT2 and EAAT3 in a fluorescence-based membrane potential assay. AB - We have expressed the human excitatory amino acid transporters EAAT1, EAAT2 and EAAT3 stably in HEK293 cells and characterized the transporters pharmacologically in a conventional [(3) H]-d-aspartate uptake assay and in a fluorescence-based membrane potential assay, the FLIPR Membrane Potential (FMP) assay. The K(m) and K(i) values obtained for 12 standard EAAT ligands at EAAT1, EAAT2 and EAAT3 in the FMP assay correlated well with the K(i) values obtained in the [(3) H]-d aspartate assay (r(2) values of 0.92, 0.92, and 0.95, respectively). Furthermore, the pharmacological characteristics of the cell lines in the FMP assay were in good agreement with previous findings in electrophysiology studies of the transporters. The FMP assay was capable of distinguishing between substrates and non-substrate inhibitors and to discriminate between "full" and "partial" substrates at the transporters. Taking advantage of the prolific nature of the FMP assay, interactions of the EAATs with substrates and inhibitors were studied in some detail. This is the first report of a high throughput screening assay for EAATs. We propose that the assay will be of great use in future studies of the transporters. Although conventional electrophysiology set-ups might be superior in terms of studying sophisticated kinetic aspects of the uptake process, the FMP assay enables the collection of considerable amounts of highly reproducible data with relatively little labor. Furthermore, considering that the number of EAAT ligands presently available is limited, and that almost all of these are characterized by low potency and a low degree of subtype selectivity, future screening of compound libraries at the EAAT-cell lines in the FMP assay could help identify structurally and pharmacologically novel ligands for the transporters. PMID- 15135309 TI - Molecular cloning and characterization of a glutathione S-transferase from largemouth bass (Micropterus salmoides) liver that is involved in the detoxification of 4-hydroxynonenal. AB - We are currently investigating the role of detoxification pathways in protecting against the sublethal effects of chemicals in largemouth bass (Micropterus salmoides). To this end, previous work in our laboratory indicated a remarkable ability of bass liver glutathione S-transferases (GSTs) to detoxify 4 hydroxynonenal (4HNE), a common mutagenic and cytotoxic alpha,beta-unsaturated aldehyde produced during the peroxidation of lipids. In the current study, we observed that GST-mediated 4HNE conjugation in bass liver follows high efficiency single-enzyme Michaelis-Menten kinetics, suggesting that an individual GST isoform is involved in 4HNE detoxification. Using 5' and 3' rapid amplification of cDNA ends (RACE), a full-length GST cDNA of 957 base pairs (bp) in length, containing an open reading frame of 678 bp and encoding a polypeptide of 225 amino acids, has been cloned. Interestingly, a search of the BLAST protein database revealed the presence of homologous GST proteins in the plaice (Pleuronectes platessa), European flounder (Platichthys flesus) and fathead minnow (Pimephales promelas), but not in other fish species. Furthermore, the bass GST protein exhibited little homology with the mammalian GSTA4 subclass of proteins which rapidly metabolize 4HNE. The recombinant 6 x His-tagged expressed GST protein showed high catalytic activity towards 4HNE, while showing moderate or low activity toward other class specific GST substrates. HPLC-GST subunit analysis, followed by sequencing, demonstrated that the isolated bass liver GST subunit constitutes the major GST protein in bass liver, with a molecular mass of 26.4 kDa. In summary, the presence of a highly expressed GST isozyme in bass and several evolutionarily divergent fish species indicates the conservation of an important and distinct detoxification protein that protects against oxidative damage in certain aquatic organisms. PMID- 15135310 TI - Inverse gene expression patterns for macrophage activating hepatotoxicants and peroxisome proliferators in rat liver. AB - Macrophage activation contributes to adverse effects produced by a number of hepatotoxic compounds. Transcriptional profiles elicited by two macrophage activators, LPS and zymosan A, were compared to those produced by 100 paradigm compounds (mostly hepatotoxicants) using cDNA microarrays. Several hepatotoxicants previously reported to activate liver macrophages produced transcriptional responses similar to LPS and zymosan, and these were used to construct a gene signature profile for macrophage activators in the liver. Measurement of cytokine mRNAs in the same liver samples by RT-PCR independently confirmed that these compounds are associated with macrophage activation. In addition to expected effects on acute phase proteins and metabolic pathways that are regulated by LPS and inflammation, a strong induction was observed for many endoplasmic reticulum-associated stress/chaperone proteins. Additionally, many genes in our macrophage activator signature profile were well-characterized PPARalpha-induced genes which were repressed by macrophage activators. A shared gene signature profile for peroxisome proliferators was determined using a training set of clofibrate, WY 14643, diethylhexylphthalate, diisononylphthalate, perfluorodecanoic acid, perfluoroheptanoic acid, and perfluorooctanoic acid. The signature profile included macrophage activator-induced genes that were repressed by peroxisome proliferators. NSAIDs comprised an interesting pharmacological class in that some compounds, notably diflunisal, co-clustered with peroxisome proliferators whereas several others co-clustered with macrophage activators, possibly due to endotoxin exposure secondary to their adverse effects on the gastrointestinal system. While much of these data confirmed findings from the literature, the transcriptional patterns detected using this toxicogenomics approach showed relationships between genes and biological pathways requiring complex analysis to be discerned. PMID- 15135312 TI - Nicotinamide as a potential addition to the anti-atopic dermatitis armamentarium. PMID- 15135311 TI - WIF-B cells as a model for alcohol-induced hepatocyte injury. AB - A potential in vitro model for studying the mechanisms of alcohol-induced hepatocyte injury is the WIF-B cell line. It has many hepatocyte-like features, including a differentiated, polarized phenotype resulting in formation of bile canaliculi. The aim of this study was to examine the effects of ethanol treatment on this cell line. WIF-B cells were cultured up to 96 h in the absence or presence of 25 mM ethanol and subsequently were analyzed for ethanol-induced physiological and morphological changes. Initial studies revealed WIF-B cells exhibited alcohol dehydrogenase (ADH) activity, expressed cytochrome p4502E1 (CYP2E1), and efficiently metabolized ethanol in culture. This cell line also produced the ethanol metabolite acetaldehyde and exhibited low K(m) aldehyde dehydrogenase (ALDH) activity, comparable to hepatocytes. Ethanol treatment of the WIF-B cells for 48 h led to significant increases in the lactate/pyruvate redox ratio and cellular triglyceride levels. Ethanol treatment also significantly altered WIF-B morphology, decreasing the number of bile canaliculi, increasing the number of cells exhibiting finger-like projections, and increasing cell diameter. The ethanol-induced changes occurring in this cell line were negated by addition of the ADH inhibitor, 4-methylpyrazole (4-MP), indicating the effects were due to ethanol metabolism. In summary, the WIF-B cell line metabolizes ethanol and exhibits many ethanol-induced changes similar to those found in hepatocytes. Because of these similarities, WIF-B cells appear to be a suitable model for studying ethanol-induced hepatocyte injury. PMID- 15135314 TI - Effects of marine beta-1,3 glucan on immune reactions. AB - Glucans have a long history as nonspecific biological modulators. A novel glucan Phycarine-was isolated from sporophytes of Laminaria digitata. Phycarine showed significant stimulation of phagocytic activity as well as potentiation of synthesis and release of IL-1, IL-6 and TNF-alpha. In addition, Phycarine increased NK cell-mediated killing of tumor cells both in vitro and in vivo while acting via complement receptor type 3 (CR3) receptors. PMID- 15135313 TI - Differential regulation of resveratrol on lipopolysacchride-stimulated human macrophages with or without IFN-gamma pre-priming. AB - Resveratrol, a polyphenol compound found in grapes and red wines, is a prominent anti-cancer agent. In this study, we demonstrate that resveratrol enhanced TNF alpha, IL-12 and IL-1beta production from LPS activated phorbol myristate acetate (PMA) differentiated THP-1 human macrophages. Expression of CD86 on macrophages was enhanced by resveratrol alone and with LPS. When macrophages were primed with IFN-gamma, resveratrol suppressed the expression of HLA-ABC, HLA-DR, CD80, CD86 and inhibited production of TNF-alpha, IL-12, IL-6 and IL-1beta induced by LPS. The differential impact of resveratrol on expression of CD14 might be correlated with differential response of macrophages to LPS with or without IFN-gamma priming. PMID- 15135315 TI - Construction of an immunotoxin with the pore forming protein StI and ior C5, a monoclonal antibody against a colon cancer cell line. AB - Sticholysin I (StI), a potent cytolysin isolated from the sea anemone Stichodactyla helianthus, was linked to the monoclonal antibody (mAb) ior C5. StI acts by forming hydrophilic pores in the membrane of the attacked cells leading to osmotic lysis. ior C5 is a murine IgG1, which recognizes the tumor associated antigen (TAA) ior C2. The cytolysin and the mAb were coupled by using the heterobifunctional cross-linking reagent sulfosuccinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC). Two hybrid molecules composed by one ior C5 and one or two StI molecules were obtained (named conjugated I and II, respectively). The purified conjugates were evaluated by a binding affinity assay against an ior C2-positive colon cancer cell line (SW948). Both molecules were able to recognize the antigen (Ag) in the same way that unconjugated ior C5 does. The activity of both conjugates against human erythrocytes and SW948 cells was assessed. They lost most of their hemolytic activity but their residual activity was very similar. Nevertheless, when their cytotoxicity was studied on the SW948 cell line, only conjugate II killed efficiently the cells, indicating a specific mAb Ag interaction. In this chimeric molecule the ratio between the cytotoxic and the hemolytic activity was larger than that of the free cytolysin. This fact indicates an increase of the specificity of the toxic effect toward the SW948 cell line and consequently an increase of the difference between its hemolytic and cytotoxic doses. The results herein support the feasibility of directing StI to the surface of cancer cells expressing ior C2 Ag via the mAb ior C5. PMID- 15135316 TI - Immunoglobulin therapy to control lung damage in patients with common variable immunodeficiency. AB - BACKGROUND: Lung damage progression is the most frequent condition in patients with common variable immunodeficiency (CVID). Appropriate immunoglobulin dose adjustments and follow-up guidelines to evaluate this have not been well established. OBJECTIVE: To assess the evolution of lung damage once stable residual serum levels of IgG over 600 mg/dl had been achieved. METHODS: A prospective study was conducted in 24 adult patients consecutively diagnosed with CVID, with no previous intravenous immunoglobulin (IVIG) treatment. IVIG dose, total serum IgG level, bacterial infection rate, pulmonary function tests (PFTs) and high resolution computed tomography (HRCT) of the thorax were monitored over 2 years. Moreover, outcome data were determined by measurement of chronic pulmonary disease (CPD). RESULTS: IVIG dose variability (205-372 mg/kg/21 days) to obtain the required serum IgG levels was determined. Patients with CPD needed higher doses than those without CPD (p=0.045). A significant reduction in severe and mild infections/patient-year was observed during treatment. Overall, there were no changes in PFTs and HRCT scores in patients without CPD, but both improved in patients with CPD. An increase of over 15% in overall HRCT score was detected in two patients without evidence of impairment in either clinical status or PFT values. CONCLUSIONS: Residual levels of total IgG over 600 mg/dl may help prevent progression of lung damage in patients with CVID. Levels of IgG, clinical manifestations and PFTs seem sufficient for routine follow-up. HRCT examination of the thorax, at least biennially, may help to identify patients in whom lung injury is progressing even though they may remain symptom-free and with stable PFTs. PMID- 15135317 TI - Increased levels of cyclic adenosine monophosphate contribute to the hyporesponsiveness of mast cells in alloxan diabetes. AB - In this study, we investigated the influence of intracellular cyclic adenosine monophosphate (cAMP) changes on the rat mast cell hyporesponsiveness following immunological and non-immunological stimuli. Compared with mast cells from normal rats, those recovered from 21-day diabetic animals showed a significant augmentation in the intracellular levels of cAMP, in directly correlated with secretion of lower amounts of histamine after stimulation with antigen, bradykinin and compound 48/80 in vitro. Incubation of normal mast cells with selective inhibitors of phosphodiesterase type 4 (PDE 4) rolipram, NCS 613 and RP 73401, or the cell permeable analogue N6-2'-O-dibutyryladenosine 3':5'-cyclic monophosphate (db cAMP), led to a decrease of histamine secretion in vitro. However, the effectiveness of either NCS 613 or db cAMP in inhibiting antigen induced degranulation is comparable in both normal and diabetic mast cells. We suggest that (a) there is a close correlation between higher levels of intracellular cAMP and hyporesponsiveness of diabetic mast cells, phenomena probably associated with a reduction in the expression and/or activity of PDE 4 and that (b) the mechanism of cAMP-mediated down-regulation of mast cell function is saturated in diabetic rats. PMID- 15135318 TI - Expression profiles of genes involved in the mouse nuclear factor-kappa B signal transduction pathway are modulated by mangiferin. AB - The polyphenol mangiferin (MA) has been shown to have various effects on macrophage function, including inhibition of phagocytic activity and of free radical production. To further characterize the immunomodulatory activity of MA, this study investigated its effects on expression by activated mouse macrophages of diverse genes related to the NF-kappaB signaling pathway, using a DNA hybridization array containing 96 NF-kappaB-related genes and on cytokine levels using a cytokine protein array. MA at 10 microM significantly inhibited the expression of (a) two genes of the Rel/NF-kappaB/IkappaB family, RelA and RelB (=I-rel), indicating an inhibitory effect on NF-kappaB-mediated signal transduction; (b) TNF receptor-associated factor 6 (Traf6), indicating probable blockage of activation of the NF-kappaB pathway by lipopolysaccharide (LPS), tumor necrosis factor (TNF), and interleukin 1 (IL-1); (c) other proteins involved in responses to TNF and in apoptotic pathways triggered by DNA damage, including the TNF receptor (TNF-R), the TNF-receptor-associated death domain (TRADD), and the receptor interacting protein (RIP); (d) the extracellular ligand IL-1alpha, again indicating likely interference with responses to IL-1; (e) the pro-inflammatory cytokines IL-1, IL-6, IL-12, TNF-alpha and RANTES (CCL5), and cytokines produced by monocytes and macrophages, including granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM CSF), macrophage colony-stimulating factor (M-CSF); (f) other toll-like receptor proteins (in addition to Traf6), including JNK1, JNK2 and Tab1; (g) Scya2 (small inducible cytokine A2=monocyte chemoattractant protein 1); and (h) various intracellular adhesion molecules (ICAMs), and the vascular cell adhesion molecule VCAM-1, which is locally increased in atheromas. The inhibition of JNK1, together with stimulation of c-JUN (i.e. the Jun oncogene) and the previously reported superoxide-scavenging activity of MA, suggests that MA may protect cells against oxidative damage and mutagenesis. Taken together, these results indicate that MA modulates the expression of a large number of genes that are critical for the regulation of apoptosis, viral replication, tumorogenesis, inflammation and various autoimmune diseases, and raise the possibility that it may be of value in the treatment of inflammatory diseases and/or cancer. PMID- 15135319 TI - PGE2 induces IL-1beta gene expression in mouse osteoblasts through a cAMP-PKA signaling pathway. AB - Prostaglandin E2 (PGE2), an abundant eicosanoid in bone, has been implicated in a number of pathological states associated with bone loss and is also known to stimulate matrix metalloproteinase-1 synthesis and secretion in rat and human osteoblast cells, although the intracellular reactions responsible for this remain unclear. Interleukin-1beta (IL-1beta) is a cytokine that plays a critical role in bone remodeling and appears to act as a downstream effector of most bone resorbing agents. However, the issue of whether PGE2 regulates the expression of IL-1beta in mouse osteoblasts has not been resolved. In this work, we demonstrate that PGE2 is a potent inducer of IL-1beta production by fetal osteoblasts and show that PGE2 stimulates the activity of the IL-1beta promoter in osteoblasts, suggesting that PGE2 controls IL-1beta gene expression at least at the transcriptional level. PGE2 was found to induce IL-1beta mRNA expression in the cells within 4 h and the level of expression was maintained for 36 h. A dose related increase in IL-1beta production was found with 0.1-2.0 microM PGE2. The induction of IL-1beta protein in the medium paralleled the induction of IL-1beta mRNA levels. The role of cAMP activation in PGE2-mediated IL-1beta production was examined by the effects of forskolin, an adenylyl cyclase (AC) activator and dideoxyadenosine (DDA), an AC inhibitor. Forskolin enhanced and DDA blocked the production of IL-1beta by PGE2. In addition, PGE2-mediated IL-1beta induction was completely inhibited by the cAMP antagonist, Rp-cAMP, and protein kinase A (PKA) inhibitors of KT5720 and H89. The PGE2-induced production of IL-1beta was also blocked by the PKA inhibitor PKI14-22. However, a specific inhibitor of protein kinase C, calphostin C, had no affect on PGE2-induced IL-1beta gene expression. Among the potential agonists, forskolin was a potent inducer of IL-1beta expression, while phorbol myristate acetate and serum had little effect. These findings indicate that PGE2 involves the cAMP-PKA signaling pathway in regulating IL-1beta gene expression in osteoblasts. PMID- 15135320 TI - Administration of macrophage colony-stimulating factor mobilized both CD11b+CD11c+ cells and NK1.1+ cells into peripheral blood. AB - We attempted the phenotypic characterization of peripheral blood (PB) cells after daily administration of macrophage colony-stimulating factor (M-CSF) in mice. The number of CD11b+ cells was increased by M-CSF treatment (2- and 5-day injections). Notably, CD11bbrightCD11cdim, CD11b+CD11c+ and CD11b+CD80+ cells were significantly increased by 2-day treatment of M-CSF. On the other hand, the number of NK1.1+ cells was not changed by the 2-day treatment, but it was significantly increased by the 5-day treatment. However, the numbers of CD3+ and NK1.1+CD3+ cells were not changed by M-CSF treatment. Then, mononuclear cells (MNCs) were separated from the PB of mice treated with saline or M-CSF, and they were incubated with GM-CSF + IL-4 or IL-2. Compared with the saline-treated one (S-MNCs), the MNCs of M-CSF-treated mice (M-MNCs) showed strong proliferation by the GM-CSF + IL-4 stimulation. The MNCs could stimulate proliferation of allo-T cells in the mixed lymphocyte reaction (MLR), especially the M-MNCs showed strong reaction. On the other hand, the stimulation by IL-2 induced strong cell growth of MNCs. And M-CSF treatment enhanced this response. Furthermore, the M-MNCs (stimulated by IL-2 in vitro) exhibited greater cytotoxicity against Yac-1 cells than the S-MNCs. In conclusion, we found that administration of M-CSF mobilized CD11b+, CD11b+CD11c+, CD11b+CD80+, and NK1.1+cells into PB. And the injection of M-CSF facilitates the generation of dendritic and natural killer cells from PB cells in vitro. These results suggest that the mobilized cells may provide for application of immunotherapy. PMID- 15135321 TI - Signaling via the AHR leads to enhanced usage of CD44v10 by murine fetal thymic emigrants: possible role for CD44 in emigration. AB - Signaling via the endogenous arylhydrocarbon receptor (AHR) affects proliferation, differentiation, function and gene expression of thymocytes. In the present study, we show that treatment of mouse fetal thymus lobes in organ culture (FTOC) with AHR ligands results in (a) a drastic decrease in the emigration of thymocytes in terms of numbers and types of cells, and (b) preferential emigration of CD4-CD8- (DN) cells expressing CD44v7- and CD44v10 containing isoforms on the cell surface. Moreover, a higher level of transcripts of various other CD44 variant isoforms (CD44v) could be detected by RT-PCR in emigrants from fetal thymi exposed to either AHR-agonist during culture. Expression of CD44v9-10-containing isoforms could be exclusively detected in DN thymic emigrants. Thus, signaling via AHR by ligands alters CD44v expression patterns in a thymocyte subpopulation. Furthermore, emigration could be decreased by the addition of anti-panCD44 antibodies to TCDD-treated FTOCs, suggesting a role for CD44 in emigration. PMID- 15135322 TI - Inhibitory effect of furosemide on activation of human peripheral blood polymorphonuclear leukocytes stimulated with n-formyl-methionyl-leucyl phenylalanine. AB - The clinical efficacy of inhalatory furosemide (Fu) has been extensively studied in bronchial asthma patients but there are only a few studies addressing its action on cells participating in the underlying inflammatory process. Therefore, we investigated the effect of Fu on human peripheral blood polymorphonuclear leukocytes (PMNL) at concentrations that can be achieved in the bronchial lining fluid by inhalation, i.e. 10(-5), 10(-4) and 10(-3) M. The influence of Fu on the following PMNL parameters was investigated: intracellular calcium changes ([Ca2+]i) as a part of signal transduction and luminol dependent chemiluminescence (LCL) as an indirect measure of NADPH-oxidase activation upon n formyl-methionyl-leucyl-phenylalanine (fMLP) stimulation; chemotaxis to fMLP, phagocytosis and intracellular killing of Staphylococcus aureus. Incubation with Fu resulted in a concentration dependent reduction of Ca2+ influx and Fu (10(-3) M) decreased the main Ca2+ parameters to one half of the control values and to the level obtained in calcium-free buffer. In contrast, Fu had no effect if preincubated with the cells and then removed by washing. The LCL signal was reduced by Fu (10(-3) M) from 2000 +/- 870 to 550 +/- 440 arbitrary units [aU] (p<0.05). In contrast to the [Ca2+]i measurements, a slightly diminished LCL was also observed following preincubation with Fu and washing. No effect of Fu was found on phagocytosis and intracellular killing of St. aureus. Fu diminished chemotaxis to fMLP but at 10(-3) M it also displayed weak chemoattractant properties. The differential action of Fu on human PMNL may add to the understanding of its topical and restricted efficacy in bronchial asthma. PMID- 15135323 TI - PADMA 28 modulates interferon-gamma-induced tryptophan degradation and neopterin production in human PBMC in vitro. AB - Tibetan herbal remedy PADMA 28 revealed promising results to support treatment of intermittent claudication, atherosclerosis and chronic hepatitis. The remedy was confirmed to be closely linked with anti- and pro-oxidative properties in vitro. In this study, effect of PADMA 28 was investigated in stimulated and unstimulated human peripheral blood mononuclear cells (PBMC) in vitro. Neopterin production and tryptophan degradation were measured in supernatants of PBMC in the presence or absence of mitogens phytohaemagglutinin (PHA) and concanavalin A (Con A). Stimulation of PBMC induced neopterin formation and tryptophan degradation (p<0.001 compared to unstimulated PBMC), and PADMA 28 inhibited both immunobiochemical effects (p<0.001) in a concentration-dependent manner. Higher concentrations of PADMA 28 were more effective and were able to completely block the pathways induced upon mitogenic stimulation. Data allow to conclude that PADMA 28 is able to inhibit immunobiological effects in stimulated PBMC in vitro. The suppression of neopterin production and tryptophan degradation suggests a specific influence on biochemical pathways induced by Th1-type cytokine interferon-gamma. PMID- 15135324 TI - Immune response modulation to DPT vaccine by aqueous extract of Withania somnifera in experimental system. AB - Immunopotentiation on oral feeding of standardized aqueous extract of Withania somnifera (Linn. Dunal, Family Solanaceae) was evaluated in laboratory animals immunized with DPT (Diphtheria, Pertussis, Tetanus) vaccine. The immunostimulation was evaluated using serological and hematological parameters. Treatment of immunized animals with test material (100 mg/kg/day) for 15 days resulted in significant increase of antibody titers to B. pertussis (P=0.000007). Immunized animals (treated and untreated) were challenged with B. pertussis 18,323 strain and the animals were observed for 14 days. Results indicate that the treated animals did show significant increase in antibody titers as compared to untreated animals after challenge (P=0.000003). Immunoprotection against intracerebral challenge of live B. pertussis cells was evaluated based on degree of sickness, paralysis and subsequent death. Reduced mortality accompanied with overall improved health status was observed in treated animals after intracerebral challenge of B. pertussis indicating development of protective immune response. Present study indicates application of the test material as potential immunopotentiating agent possible applications in immunochemical industry. The test material also offers direct therapeutic benefits resulting in reduced morbidity and mortality of experimental animals. PMID- 15135325 TI - Integrin beta 4 is a target of rattlesnake venom during inducing apoptosis of vascular endothelial cells. AB - To find more effective components which can trigger apoptosis in crude rattlesnake venom, and the possible mechanisms by which the venom causes apoptosis in vascular endothelial cells (VECs), we investigated the function of integrin beta4 by using the monoclonal antibody (mAb) of this integrin. We added anti-beta4 mAb 5 microg.ml(-1) to the cells treated with 2 microg.ml(-1) rattlesnake venom; apoptosis of these cells was completely inhibited 6 h after the treatment. Furthermore, the increase of P53 expression induced by the venom was markedly suppressed. The results first demonstrated that there was at least one important component target to integrin beta4 in crude rattlesnake (Crotalus atrox) venom; moreover, this component played its role in the early phase of apoptosis. The results also showed that integrin beta4 participated in signal transduction of apoptosis induced by rattlesnake venom in VEC by up-regulating the expression of p53. PMID- 15135326 TI - Effects of R-102444, an orally active 5-HT2A receptor antagonist, in rat models of peripheral vascular disease. AB - R-102444 is a prodrug that is metabolized into R-96544, a potent and selective 5 hydroxytryptamine2A (5-HT2A) receptor antagonist. The effects of R-102444 on peripheral vascular disease were examined using two different rat models: one induced by lauric acid and the other by ergotamine plus epinephrine. R-96544 (0.3 30 nM) relaxed the 5-HT (3 microM)-precontracted rat caudal artery in a concentration-dependent manner. The intravenous administration of R-96544 (0.3-3 microg/kg) to anesthetized rats inhibited the pressor response to 5-HT (50 microg/kg i.v.) dose dependently. The oral administration of R-102444 (1 mg/kg) to rats resulted in a marked inhibition of platelet aggregation induced by 5-HT plus ADP, and statistically significant inhibition was still evident 8 h after the dosing. In contrast, sarpogrelate, at a dose of 100 mg/kg p.o., produced only a moderate antiplatelet effect. Oral administration of R-102444 (1 mg/kg/day, o.d.) significantly prevented the progression of peripheral vascular lesion induced by the injection of lauric acid into a rat femoral artery, whereas sarpogrelate (100 mg/kg/day) showed only a minimal effect. Both 5-day treatments with R-102444 (1-30 mg/kg/day p.o., o.d.), one commenced 1 h before the injection of epinephrine plus ergotamine and one just after injection, resulted in the prevention of rat tail gangrene in a dose-dependent manner, whereas sarpogrelate (100 mg/kg) produced a minimal protection in this model. Based on these results, we conclude that 5-HT2A receptor activation is involved in peripheral vascular disease in the rat and that R-102444 is a useful oral agent for the investigation of diseases involving 5-HT2A receptor activation. PMID- 15135327 TI - Study of correlation between elevation of blood pressure and tissue ACE activity during development of hypertension in 1K1C rats. AB - OBJECTIVES: This study sought to examine the alteration of local angiotensin converting enzyme (ACE) activity in the aortae, heart, kidney and lung as well as plasma during the development of hypertension in one-kidney, one-clip (1K1C) model, a non-renin-dependent model of renovascular hypertension. METHODS: Experiments were carried out 2, 4, 8 and 12 weeks after induction of hypertension in male Sprague-Dawley rats. ACE activity was analyzed by high-performance liquid chromatography (HPLC) and the structural changes in aortae were investigated by measurement of cross-sectional area (CSA). RESULTS: Our results show that ACE activity in aortae and heart was gradually increased with the development of hypertension and was more pronounced at higher blood pressure. In addition, there was a positive correlation between aortic CSA and elevation of blood pressure. CONCLUSIONS: Our findings emphasize the significant role of local ACE, particularly in organs regulating hypertension (aortae and heart) in 1K1C model, in which circulatory renin is known to be unelevated. PMID- 15135328 TI - Diphenylamine-2-carboxylic acid potentiates the cyclic nucleotides-mediated relaxation of porcine coronary artery: possible involvement of the inhibitory effect on the efflux of cyclic nucleotides. AB - We examined the effect of diphenylamine-2-carboxylic acid (DPC), which has been shown to inhibit the efflux of cyclic nucleotides from vascular smooth-muscle cells, on the relaxant responses to forskolin, an adenylyl cyclase activator, and sodium nitroprusside (SNP), an NO donor, in the porcine coronary arteries. DPC (100 microM), which caused only a minor effect by itself, significantly augmented the relaxant responses to forskolin and SNP in the preparations contracted with 30 mM KCl. On the other hand, DPC did not affect the relaxant responses to nifedipine and cromakalim. Forskolin (10 microM) induced an accumulation of adenosine 3', 5'-cyclic monophosphate (cAMP) in the porcine coronary arteries, which was associated with an accumulation of cAMP in the incubation media. The intracellular cAMP response to forskolin was enhanced by DPC, whereas the extracellular cAMP response was reduced. The effects of SNP on guanosine 3', 5' cyclic monophosphate (cGMP) accumulation were examined in the presence of 3 isobutyl-l-methylxanthine (500 microM) because cGMP was not found in the tissue and the incubation medium in the absence of the phosphodiesterase inhibitor. DPC significantly decreased the SNP-induced release of cGMP to the extracellular space, whereas it did not affect the accumulation of cGMP in the tissue. These results suggest that DPC inhibits the efflux of cyclic nucleotides. It is likely that the inhibitory effect of DPC on cAMP efflux contributes to the enhancement of tissue cAMP accumulation and relaxation produced by the agents that activate adenylyl cyclase. Thus, the transport system(s) of cyclic nucleotides may be a novel target for the prevention and/or treatment of various cardiovascular diseases. PMID- 15135329 TI - Effect of single oral administration of YM598, a novel selective endothelin ETA receptor antagonist, on blood pressure in normotensive and hypertensive rats. AB - We investigated the effect of YM598, a selective endothelin ETA receptor antagonist, on blood pressure (BP) in normotensive rats (NTR), spontaneously hypertensive rats (SHR) and Dahl salt-sensitive hypertensive rats (Dahl-SS). We also examined the concomitant effect of YM598 with the L-type Ca2+ channel antagonist nifedipine on BP. Single oral administration of YM598 did not affect BP in NTR and SHR. In Dahl-SS, in contrast, YM598 slightly, but not significantly, reduced BP. Concomitant administration of YM598 with nifedipine at doses inducing slight hypotension on respective single administrations resulted in a stronger hypotensive effect than single administration of either compound alone. However, the magnitude of the concomitant hypotensive effect demonstrated only a simple additive effect of the two compounds. These results indicate that YM598 did cause slight hypotensive effects in some types of hypertension. These results also indicate the possibility of additive, but not synergic, hypotensive effects on concomitant administration of ET receptor antagonist and an L-type Ca2+ channel antagonist. PMID- 15135331 TI - Ethnic differences in preferences for female weight and waist-to-hip ratio: a comparison of African-American and White American college and community samples. AB - This study examined both ethnic differences and differences between college and community samples in male preferences for ideal body size and shape in women. As expected, it was found that African-American men were more likely to choose heavier figures as ideal than White American men did. Contrary to expectations, there were no differences in chosen ideal weight between college and community subsamples. As expected, both ethnic groups chose figures with a low waist-to-hip ratio (WHR), but African-American men were more likely to choose a very low WHR as ideal. The findings replicate and extend research showing weight to be a more important cue than WHR in the mate-selection process and support the theory that African-American men's preferences may serve as a protective factor against eating and body image pathology in African-American women. PMID- 15135330 TI - Antiplatelet and antithrombotic activities of CP201, a newly synthesized 1,4 naphthoquinone derivative. AB - The antiplatelet and antithrombotic activities of a newly synthesized CP201, 2 (3,5-di-tert-butyl-4-hydroxyl)-3-chloro-1,4-naphthoquinone on human platelet aggregation in vitro and murine pulmonary thrombosis in vivo were examined. In addition, the antiplatelet activity of CP201 involved in calcium-signaling cascade was also investigated. CP201 showed concentration-dependent inhibitory effects on platelet aggregation induced by collagen and thrombin, with IC50 values of 4.1+/-0.3 and 4.6+/-0.4 microM, respectively. Orally administered CP201 protected mice against the collagen plus epinephrine-induced thromboembolic death in a dose-dependent manner. On the other hand, CP201 did not alter such coagulation parameters as activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) in human plasma in vitro. These results suggest that the antithrombotic activity of CP201 may be due to antiplatelet rather than anticoagulation activity. CP201 potently inhibited platelet aggregation challenged by calcium ionophore A23187 and thapsigargin, which is a selective inhibitor of the Ca(2+)-ATPase pump, in a concentration dependent manner, indicating that CP201 may have an inhibitory effect on calcium signaling cascade. This was supported by measuring [Ca2+]i in platelets loaded with fura-3AM, where CP201 inhibited the rise in cytosolic Ca2+ mediated by thrombin. Taken together, these results suggest that CP201 may be a promising antithrombotic agent, and the antithrombotic effect of CP201 may be due to antiplatelet activity, which was mediated, at least partly, by the inhibition of cytosolic calcium mobilization. PMID- 15135332 TI - What a difference a diet makes: towards an understanding of differences between restrained dieters and restrained nondieters. AB - Restrained eaters who are and are not dieting to lose weight have shown opposite eating regulation patterns in past research. To better understand these differences, restraint theory and the Three-Factor Model of Dieting was used to generate differential predictions about the mean and variability of restrained dieters (RDs) and restrained nondieters (RNDs) on the eating inventory Cognitive Restraint (CR) scale and the Restraint Scale (RS). Unrestrained nondieters served as a reference group. Eighty normal-weight female college students completed the CR, RS, and a measure of weight cycling. RDs, relative to RNDs, obtained higher and more homogeneous scores on the Cognitive Restraint, and higher and more heterogeneous scores on the RS. A post hoc analysis found that RDs had a much greater weight cycling history than RNDs. These findings are most consistent with the Three-Factor Model of Dieting, but also point to needed revisions both in this model and in traditional restraint theory. PMID- 15135333 TI - Similarities and differences between women and men on eating disorder risk factors and symptom measures. AB - Researchers studying eating disorders in men often use eating disorder risk and symptom measures that have been validated only on women. Using a sample of 215 college women and 214 college men, we report on the validity of doing so with a set of measures chosen to reflect a wide range of risk factors and symptoms. The Bulimia Test-revised (BULIT-R), the restraint scale (RS), the three-factor eating questionnaire (TFEQ), the Eating Expectancy Inventory, and the eating attitudes test (EAT) all had the same factor structures for both genders, and tests of invariance showed that factor loadings, factor variances, and intercorrelations among factors were equivalent across gender. A modified Structured Clinical Interview for DSM-IV anorexic symptoms questionnaire did not perform adequately for either gender. Men produced slightly less reliable scores on virtually all measures, with the result that Pearson-based estimates of correlations among the measures were slightly lower for men. Men had lower scores on symptom and risk measures, but not on other eating measures. PMID- 15135334 TI - An experimental analysis of the role of schema compensation in anorexia nervosa. AB - It has been suggested that the relatively poor effectiveness of treatments for anorexia nervosa is due to a poor conceptualisation of the disorder. One hypothesis is that current models are mistakenly targeting superficial, instead of deeper level, cognitions and cognitive processes. A schema-based cognitive behavioural model of eating disorder pathology suggests that the process of schema compensation is key to restrictive pathology-when there is the threat of experiencing negative affect, compensatory schemas are activated, reducing that affect. The current experimental study aimed to provide support for such a process. Eating-disordered and control women completed a computer-based task, measuring the compensation process in terms of speed and accuracy in response to subliminal threat cues. The results did not fully support the hypothesis, suggesting that the model and methodology need some amendment. Improvements to the methodology are discussed. PMID- 15135335 TI - Chewing and spitting in eating disorders and its relationship to binge eating. AB - OBJECTIVE: This study examined (i) the frequency of chewing and spitting and (ii) its association with other pathological eating behaviors in eating-disordered inpatients. We hypothesized a positive association between chewing and spitting and binge eating given the phenomenological similarities between these disordered eating behaviors. METHOD: Frequent chewers/spitters were compared with those who did not regularly engage in this behavior with regard to diagnosis, psychometric test results, and associated eating pathology. RESULTS: Chewing and spitting was not associated with elevated bingeing. Rather, frequent chewers/spitters exhibited higher levels of restrictive eating behaviors and the behavior was more prevalent in younger patients. DISCUSSION: Contrary to our predictions, chewing and spitting is more closely associated with restrictive than with binge behaviors. This suggests that most individuals chew and spit small portions of food. The behavior is frequent, occurs across diagnostic groups, and may be associated with greater psychopathology. Future studies should clarify the amount of food consumed during chew/spit episodes and the presence of a sense of loss of control. PMID- 15135336 TI - Energy and macronutrient intake in bulimia nervosa. AB - Energy deprivation and malnutrition are often thought to be key factors in the maintenance of bulimia nervosa (BN). Our review shows that it is unclear how much energy is actually available to BN patients' metabolism because the contribution of food consumed during binge eating is generally neglected. Also, there is little evidence for another key hypothesis that binge-eating episodes are triggered by carbohydrate craving. This study examined energy consumption and macronutrient composition of meals and binge-eating episodes in food diaries. Forty female BN patients, 40 female panic disorder (PD) patients, and 40 healthy women recorded their food intake while in their natural environment during two consecutive days. We did not find the expected evidence for chronic energy deprivation and malnutrition in BN patients. Also, there was no evidence that carbohydrate craving drives binge eating. The implications for models of BN and for treatments targeting eating behavior are discussed. PMID- 15135337 TI - Dietary restraint and self-esteem as predictors of weight gain over an 8-year time period. AB - The study aimed to assess dietary restraint and self-esteem as predictors of weight change over a time period of 8 years. Questionnaires assessing self reported weight, body dissatisfaction, dietary restraint, and self-esteem were completed by 77 young adults (19 men and 58 women) on two occasions separated by 8 years. On average, participants gained approximately 6 kg over the 8 years. Although neither dietary restraint nor self-esteem predicted weight change on its own, their interaction did. Furthermore, the relationship between restraint and weight change was best described as curvilinear. It was concluded that dietary restraint is predictive of subsequent weight gain, but in a more complex way than previously assumed. PMID- 15135338 TI - The role of sensitivity to external food cues in attentional allocation to food words on dot probe and Stroop tasks. AB - The role of sensitivity of external food cues in producing attentional bias toward food-, body-weight-, and shape-related words on the Stroop and the dot probe tasks was examined. Contrary to expectations, individuals high in responsiveness to external food cues directed attention away from food words, whereas individuals low in responsiveness to external food cues directed their attention toward food words on the dot probe task. No significant differences were found between the groups high and low in sensitivity to external food cues for body words on the dot probe task or for food or body words on the Stroop task. Results are discussed with reference to theoretical views of differences between the Stroop and the dot probe tasks. PMID- 15135339 TI - Tissue transglutaminase, a key enzyme involved in liver diseases. AB - Ubiquitous tissue transglutaminase (tTG) is one member of the large transglutaminase (TG) family, which catalyze posttranslational modification of proteins by establishing epsilon(gamma-glutamyl)lysine cross-linking and/or covalent incorporation of polyamines. The unique characteristics of tTG include: (1) possessing both cross-linking activity and GTPase activity; (2) functioning as a G protein; and (3) participating in the signal transduction of alpha1 adrenergic receptor coupling. A growing body of literature suggests that increased tTG levels in the cytosolic or nuclear compartments contribute to the apoptotic process, and lines of evidence exist that nuclear translocation and cross-linking of transcriptional factor Sp1 may represent the underlying mechanisms of these proapoptotic effects of tTG. Our studies indicate that tTG GTPase activation may be responsible for enhanced hepatocyte proliferation, whereas, its tTGase activity may cause increased apoptosis. Moreover, it appears that tTG cross-linking activity contributes to hepatic fibrogenesis in animal models and in human liver disease. Understanding the roles of tTG in the pathogenesis of liver disease could facilitate the development of new treatment regimens. PMID- 15135340 TI - A randomized controlled trial of consensus interferon with or without lactoferrin for chronic hepatitis C patients with genotype 1b and high viral load. AB - Recently, lactoferrin has been reported to have anti-HCV effects. The aim of this study was to investigate the effect of combination therapy using consensus interferon (CIFN) and lactoferrin in patients with chronic hepatitis C. Twenty one patients with chronic HCV infection, who were positive for HCV-RNA genotype 1b with serum viral loads from 100 to 700KIU/ml, were randomly assigned to two groups; the CIFN + Lac group received CIFN with lactoferrin and the CIFN group received CIFN alone. Nine patients in each group completed this trial; the other patients dropped out because of side effects. Three, two and four patients were categorized as complete responders, relapsers and non-responders, respectively, in the CIFN + Lac group, and four, one and four in the CIFN group, respectively. There was no statistically significant difference in virologic response between the two groups. During the follow up after CIFN therapy with continued lactoferrin, there were two relapsers in the CIFN + Lac group and their HCV-RNA titers before treatment were over 400KIU/ml. In conclusion, the combination therapy of CIFN and lactoferrin did not increase the response rate or prevent relapse after discontinuation of IFN. PMID- 15135341 TI - Elasticity of small artery in patient with spontaneous rupture of hepatocellular carcinoma. AB - Objective: Spontaneous rupture of hepatocellular carcinoma (HCC) is common in Asia and Africa with unclear mechanism. In our previous study, we found that the vascular injury was related to the HCC rupture. In this study, the structure of elastin around the small artery was deeply investigated to confirm our previous study. Methods: Immunohistochemical technique and transmission electron microscopy were used to study 23 specimens from ruptured HCC and 30 cases with nonruptured HCC. Results: The layer of elastin around the vascular wall was significant thicker in patients with ruptured HCC than that in nonruptured HCC. The proliferation of elastin, abnormal distribution of neutrophil elastase and degradation of collagen fibril were predominantly present in the specimens from ruptured HCC. The phenomenon that the infiltrated neutrophils from bloodstream into the vascular wall, which caused the vascular injury, can be found in specimens from ruptured HCC. The vascular injury mainly occurred in small artery. Since the damaged vessels could become stiff and weak, which would be more prone to splitting and result in hemorrhage in patients with ruptured HCC, we postulated that the vascular injury, especially the inelastic small artery, may relate the ruptured HCC. Conclusion: The vascular injury in small artery might relate to ruptured HCC. PMID- 15135342 TI - The influence on liver parenchymal function and complications of radiofrequency ablation or the combination with transcatheter arterial embolization for hepatocellular carcinoma. AB - Objectives: We investigated how radiofrequency ablation (RFA) alone or the combination with transcatheter arterial embolization (TAE-RFA) for hepatocellular carcinoma (HCC) affects the liver parenchymal function of underlying chronic liver disease to find a predictive factor for selecting appropriate candidates for RFA. Methods: In 53 HCC patients (RFA alone, 25; TAE-RFA, 28), liver laboratory tests and development of complications were monitored until 6 months after treatments. Serum albumin deteriorated within 1 month and recovered after 6 months in both groups, whereas the elevation of the Pugh score continued in RFA alone group and that of serum bilirubin continued in TAE-RFA group until 6 months. The elevation of Pugh score 6 months after RFA was significantly more frequent in patients with a high pre-treatment Pugh score (>/=8 points) than in those with a low pre-treatment Pugh score (<8 points). Complications after RFA were observed in 11 of the 53 patients: temporary complications in 5, refractory ascites in 6. Refractory ascites developed in only patients with a high pre treatment Pugh score. Conclusion: Liver parenchymal functions in patients with a low pre-treatment Pugh score were transiently deteriorated by RFA or TAE-RFA. However, in patients with a high Pugh score, RFA or TAE-RFA induces long-term deterioration of the liver parenchymal functions and causes serious complications. Therefore, patients with a Pugh score >/=8 points would not be good candidates for RFA or TAE-RFA. PMID- 15135343 TI - Preoperative assessment of multicentric occurrence in synchronous small and multiple hepatocellular carcinoma based on image-patterns and histological grading of non-cancerous region. AB - Recent studies have shown that the prognosis of patients with multicentric occurrence (MO) of hepatocellullar carcinoma (HCC) after curative resection, whether synchronous or metachronous, is significantly better than that of patients with intrahepatic metastasis (IM). We attempted to elucidate a group with high probability of MO of HCC by comparing the preoperative ultrasonography (US), computed tomography during arterial portography (CTAP) and CT arteriography (CTA) images with the histological grading of the non-cancerous region of the liver. The imaging and pathological characteristics of 97 nodules in 45 patients diagnosed as having small (<5cm) and oligonodular HCCs preoperatively were investigated. Histological diagnoses were as follows: 18 MO, 23 IM, and 4 unclassified (HCC with benign nodule). Histological grade of the liver tissue was assessed using the hepatitis activity index (HAI) score. No significant differences in tumor location and the mean size of co- or daughter nodules were noticed between the MO and IM patients. Of the 38 nodules in the MO patients, 20 were diagnosed as hyperechoic lesions on US, 20 as negative enhanced lesions on CTA and 12 as having positive portal flow on CTAP. A comparison of image-patterns between the synchronous nodules revealed that the MO group showed different images in at least one imaging technique, whereas in most IM patients similar images were produced by two or three imaging techniques. Moreover, all MO patients had an [Formula: see text], although more than half the IM patients had an [Formula: see text]. In patients with small and oligonodular HCCs, the nodules are highly likely to display MO, when showing both 1 or more different image patterns among US, CTA and CTAP and severe hepatitis such as HAI score >/=8. PMID- 15135344 TI - Detection of telomerase activity, telomerase RNA component, and telomerase reverse transcriptase in human hepatocellular carcinoma. AB - The aim of this study was to estimate the correlation between telomerase activity and the expression of human telomerase RNA component (hTERC), human telomerase reverse transcriptase (hTERT) in patients with hepatocellular carcinoma (HCC), and to analyze the influence of the nucleotide homology of the hTERC template region on telomerase activity. Six HCC patients and two chronic hepatitis patients were enrolled in this study. Telomerase activity was determined using the fluorescence-based telomeric repeat amplification protocol (TRAP) method. Quantification of hTERC and hTERTmRNA was performed using a real-time PCR method. Furthermore, a portion of the hTERC gene was amplified using nested RT-PCR methods. After sub-cloning, the nucleotide sequence of the cloned hTERC that contained the template region was determined. Telomerase activity and hTERTmRNA was detected in all cancerous tissues, while hTERC was present in both tumorous and non-tumorous lesions. The level of telomerase activity correlated with expression of hTERTmRNA, but not that of hTERC. The nucleotide sequence of cloned hTERC was similar in both tumorous and non-tumorous lesions. The expression of hTERT may be a definitive factor in the activation of telomerase in hepatocarcinogenesis. PMID- 15135345 TI - Control of oxidative stress and reduction in biochemical markers by Vitamin E treatment in patients with nonalcoholic steatohepatitis: a pilot study. PMID- 15135346 TI - Effects of insulin-like growth factor-type 1 on weight gain and hepatic glycogen during early development in a surfactant/virus mouse model of acute liver failure: correlation with mortality. AB - Acute liver failure (ALF) was reproduced in young mice exposed daily for 12 days to the industrial surfactant, Toximul 3409F (Tox), and infected on postnatal day (P) 14 with sublethal doses of mouse-adapted human influenza B (Lee) virus (FluB). Combined Tox + FluB treatment potentiated mortality due to non-necrotic ALF. This study tested the hypothesis that mortality would decline if the known losses in energy production due to compromised fatty-acid beta-oxidation were compensated by pharmacological manipulation of hepatic glycogen stores. Glycogen levels, body weights, and mortality were determined without and with injections of insulin-like growth factor-1 (IGF-1). On P25, 13 days after Tox exposure ceased, glycogen levels (mg/100mg) were: 4.0 (control), 1.7 (Tox), 4.3 (FluB), and 2.9 (Tox + FluB). Corresponding cumulative mortalities were 0, 14, 2, and 38%. Following daily IGF-1 injections from P12 to P17, liver glycogen levels on P25 were: 3.5 (IGF-1), 3.9 (IGF-1 + Tox), 12.3 (IGF-1 + FluB), and 5.6 (IGF-1 + Tox + FluB). Unexpectedly, IGF-1 treatment increased mortality to 67% (IGF-1), 89% (IGF-1 + Tox), 63% (IGF-1 + FluB), and 81% (IGF-1 + Tox + FluB). For all groups there was a significant correlation between mortality and poor weight gain. This is the first report of persistent glycogen reductions after surfactant exposure and withdrawal. Their role in potentiating FluB-induced mortality remains to be established. PMID- 15135347 TI - Angiopoietin/tie receptors system may play a role during reconstruction and capillarization of the hepatic sinusoids after partial hepatectomy and liver necrosis in rats. AB - Sinusoidal endothelial cells and hepatocytes increase in number after partial resection and necrosis of the liver, and contribute to both reconstruction and capillarization of the sinusoids through interaction with stellate cells. The mechanism underlying such interaction was evaluated regarding angiopoietins and tie receptors essential for blood vessel formation. Hepatic mRNA expressions of angiopoietin-1, angiopoietin-2 and tie-2 were increased at 168h after 70% liver resection with sinusoidal reconstruction in rats, and also at 48h with sinusoidal capillarization in the liver of rats given carbon tetrachloride (CCl(4)). Tie-2 mRNA expression was detected in sinusoidal endothelial cells and stellate cells isolated from normal rats, and in activated stellate cells from CCl(4) intoxicated rats. The mRNA expressions of angiopoietin-1 and angiopoietin-2 were detectable in Kupffer cells, sinusoidal endothelial cells and stellate cells in normal rats, but increased in activated stellate cells and macrophages after CCl(4)-intoxication. Both angiopoietins and tie-2 were immunohistochemically stained along the sinusoids of 70% hepatectomized rats and in necrotic areas of CCl(4)-intoxicated rats. Angiopoietin-1 and angiopoietin-2 may be involved in both reconstruction and capillarization of the sinusoids in rat liver after partial resection and necrosis through interaction between stellate cells and sinusoidal and vascular endothelial cells via tie-2 receptor. PMID- 15135348 TI - Modulation of organic anion transporting polypeptide 1 and multidrug resistance protein 3 expression in the liver and kidney of Gunn rats. AB - Background: Gunn rat is an animal model of Crigler-Najjar syndrome (CNS) type I that develops jaundice due to defect of bilirubin conjugation. Bilirubin UDP glucuronosyltransferase (UGT1A1), which plays a critical role in bilirubin glucuronidation, has been reported to be deficient in CNS type 1. On the other hand, little is known about the expression of organic anion transporters in Gunn rats. In the present study, we evaluated expressions of organic anion transporting polypeptide (oatp) 1 and 2, multidrug resistance-associated protein (mrp) 2 and mrp3 in the liver and kidney of Gunn rats. Methods: Serum samples, liver and kidney tissues were obtained from Gunn rats and normal SD rats ( [Formula: see text], in each group). Semi-quantitative mRNA expression of oatp1, oatp2, mrp2, and mrp3 mRNA was evaluated by constructed reverse transcription polymerase chain reaction (RT-PCR). Protein expressions were determined by Western blotting and by immunohistochemistry. Results: Marked elevation of serum unconjugated bilirubin concentration ( [Formula: see text] micromol/l) was observed in Gunn rats. Hepatic expression of oatp1 and oatp2 mRNA was 44% ( [Formula: see text] ) and 35% ( [Formula: see text] ) lower in Gunn rats than in SD rats, respectively. Hepatic oatp1 protein expression was 37% ( [Formula: see text] ) lower in Gunn rats than in SD rats. In contrast to oatp1, hepatic expression of mrp3 mRNA and protein was 76% ( [Formula: see text] ) and 557% ( [Formula: see text] ) higher in Gunn rats than in SD rats, respectively. Hepatic expression of oatp2 and mrp2 protein was not significantly different between Gunn rats and SD rats. Like the Western blot analysis, immunohistochemical staining disclosed decrease of oatp1 and increase of mrp3 protein expressions in the liver of Gunn rats. Decrease of oatp1 and increase of mrp3 expressions were also observed in the kidney of Gunn rats. Conclusion: Decreased expression of oatp1 and increased expression of mrp3 were observed in the liver and kidney of Gunn rats. Deficient UGT1A1 activity-associated retention of unconjugated bilirubin in the hepatocytes may modulate the expressions of these transporters in Gunn rats. PMID- 15135349 TI - A pilot study using synthetic feline facial pheromone for the management of feline idiopathic cystitis. AB - Synthetic feline facial pheromone (FFP) (Feliway; Ceva Animal Health) was assessed for the management of cats with recurrent feline idiopathic cystitis (FIC). Nine of 12 cats completed the randomised, double-blinded, placebo controlled, crossover pilot study. They had their environment treated daily with either FFP or placebo for 2 months, after which time the treatment groups were reversed. Owners used visual analogue scales to define the severity of their cat's clinical signs and behavioural changes. Five (56%) of the owners stated that their cat's overall health was better when they were using FFP. Four (44%) of the owners noticed no difference between when using the FFP and when using the placebo. While there were no statistical differences between the two treatment groups there was a trend for the cats exposed to FFP to show fewer days with clinical signs of cystitis (FFP total, mean per cat+/-standard deviation, 30, 4.3+/-6.7; placebo 69, 9.9+/-19.1), a lower overall clinical score (1667, 238+/ 476; 2009, 287+/-425), a reduced number of episodes of cystitis (9, 1.3+/-2.0; 10, 1.4+/-2.1) and reduced negative behavioural traits (e.g., less aggression and fear) (-128, -18.3+/-65.8; -73, -10.4+/-35.1). PMID- 15135350 TI - Whole blood transfusions in 91 cats: a clinical evaluation. AB - This survey assessed the feline transfusion practices at the University of Berlin from 1998 to 2001 in regard to patient population, indications, efficacy, and transfusion reactions. Blood was obtained from seven healthy in-house donors and 127 mostly indoor client-owned pet cats. Over a 3-year period 91 cats were transfused with blood type compatible blood. The blood was fresh (within 8 h of collection) or stored no longer than 15 days. Transfusions were required because of blood loss anaemia (n=40), haemolytic anaemia (n=13), ineffective erythropoiesis (n=35), hypoproteinaemia (n=2) or coagulopathy (n=2). The anaemic cats had a pretransfusion haematocrit of 5-20% (m [median]=13), and received one to six transfusions (m=1). The survival rates of the anaemic cats at 1 and 10 days after transfusion were 84 and 64%, respectively. None of the deaths appeared to be related to transfusion reactions. The major crossmatch, undertaken before 117 transfusions, was incompatible for eight cats. All except for one had previously been transfused. Lysis of transfused cells in six cases resulted in a less than expected haematocrit rise and an increase in serum bilirubin. Transient mild transfusion reactions were only noted in two cats during the second or third transfusion. In conclusion, with proper donor selection and appropriate compatibility screening, blood transfusions are well tolerated, appear effective, and may increase chances of survival. PMID- 15135351 TI - Investigation of serum IGF-I levels amongst diabetic and non-diabetic cats. AB - Since insulin-like growth factor-I (IGF-I) was first discovered as a mediator of glucose homeostasis, it has been extensively investigated in diabetes research in humans, rodents and primates. To date, however, relatively little work has been carried out on this hormone in the cat, despite the pathophysiological similarities between human and feline diabetes mellitus, as well as the relatively common nature of the disease in cats. This study reports on the IGF-I concentrations of 42 insulin treated diabetic cats and 25 normal cats. Diabetic subjects were grouped according to length of insulin treatment as either short, medium or long term. Analysis of variance (ANOVA) and Fischer's pair-wise comparisons revealed that mean IGF-I levels in short-term diabetic cats were significantly lower than those in normal cats whilst mean levels in long-term diabetics were significantly higher. The direction and extent of these alterations may have implications for our understanding of the pathophysiology of feline diabetes mellitus and for the use of this hormone in the diagnosis of acromegaly in diabetic cats. PMID- 15135352 TI - Metronidazole for the treatment of feline giardiasis. AB - There are several drugs available for the treatment of giardiasis in cats, including metronidazole. The purpose of this study was to determine whether metronidazole benzoate administered at a dose of 25 mg/kg, orally, twice a day for 7 days lessens or eliminates Giardia cyst shedding in cats with chronic infection. Twenty-six, adult, laboratory-reared cats were used in this study. Sixteen cats had been inoculated orally with cysts of a human Giardia sp. isolate and had completed a Giardia vaccine study in one animal holding room. The other ten cats were infected with the same Giardia sp. presumably by contamination from the adjacent room where the Giardia vaccine study cats were located. From each cat, a fecal sample was collected within 1 week of the start of treatment and then every 2 to 4 days for 15 days after treatment was completed. Fecal samples were analyzed for the presence of Giardia cysts using a commercially available direct immunofluorescence test (IFA). Clinical signs of drug toxicity were not detected during the study. PMID- 15135353 TI - The use of real-time PCR in the diagnosis and monitoring of Mycoplasma haemofelis copy number in a naturally infected cat. AB - A 5-year-old male neutered cat was diagnosed with severe anaemia due to acute Mycoplasma haemofelis infection. Inflammatory respiratory disease was present concurrently. The cat was treated successfully using a fresh transfusion of whole blood and a 6 week course of doxycycline. The patient made a rapid recovery although the allergic airway disease subsequently required specific therapy consisting of inhaled fluticasone and salbutamol. Real-time quantitative PCR assays confirmed the presence of M. haemofelis DNA copies in the blood at presentation. Repeat PCR assays showed a reduction in copy number during treatment and negative PCR results were obtained both 91 and 425 days after presentation. The report describes, for the first time, the use of real-time PCR in the diagnosis and monitoring of natural M. haemofelis copy number, as well as the induction of long-term negative PCR status. PMID- 15135355 TI - Twenty-five cases of feline bronchial disease (1995-2000). AB - Twenty-five cases of feline bronchial disease were identified retrospectively. The criteria for inclusion were consistent clinical signs or histopathology and no other identifiable aetiology. Patient records were analysed to determine historical, clinical, clinicopathologic and radiographic features. The main presenting complaints were coughing and dyspnoea. The most common physical finding was dyspnoea. The majority of radiographs had a bronchial pattern either as the sole change or as a component of a mixed pattern. Bronchoalveolar lavage cytology was neutrophilic or eosinophilic in the majority of cats. There was no association between age, breed, sex, clinical signs, bronchoalveolar lavage cytology or radiographic severity and disease severity. PMID- 15135354 TI - Lower respiratory tract infections in cats: 21 cases (1995-2000). AB - Twenty-one lower respiratory tract infections diagnosed in cats at University of Sydney Veterinary Centre between 1995 and 2000 were identified retrospectively. Patient records were analysed to determine historical, clinical, clinicopathologic and radiographic features of lower respiratory tract infections. Response to therapy was also assessed. Infectious agents identified were Mycoplasma spp., Pasteurella spp., Bordetella bronchiseptica, Salmonella typhimurium, Pseudomonas sp., Mycobacterium thermoresistible, Cryptococcus neoformans, Toxoplasma gondii, Aelurostrongylus abstrusus and Eucoleus aerophilus. The study provides a detailed retrospective analysis of infectious lower respiratory tract disease in this population of cats. PMID- 15135356 TI - A retrospective analysis of feline bronchoalveolar lavage cytology and microbiology (1995-2000). AB - A retrospective study of 88 bronchoalveolar lavage specimens from 80 cats presenting to the University of Sydney Veterinary Centre between 1995 and 2000 was performed. Bronchoalveolar lavage cytology and microbiology in conjunction with other diagnostic aids and patient records were used to classify cases as lower respiratory tract infections (LTRIs), LRTI revisits, feline bronchial disease, cardiac disease, neoplastic disease and multisystemic disease. Cases for which a definitive diagnosis could not be made were classified as inconclusive. Infectious agents identified were Mycoplasma spp., Pasteurella spp., Bordetella bronchiseptica, Salmonella typhimurium, Pseudomonas sp., Cryptococcus neoformans, Aelurostrongylus abstrusus and Eucoleus aerophilus. The study highlights the importance of bronchoalveolar lavage cytology and microbiology in the evaluation of feline lower respiratory tract disease. PMID- 15135357 TI - Treatment of a well differentiated pulmonary adenocarcinoma in a cat by pneumonectomy and adjuvant mitoxantrone chemotherapy. AB - Primary lung neoplasia is a rare condition of older cats. This report describes a case of left-sided pulmonary collapse, secondary to pulmonary adenocarcinoma in an 11-year-old cat. Left sided pneumonectomy was performed, and adjuvant chemotherapy with mitoxantrone (Novantrone; Lederle) was administered every 3 to 5 weeks, for 10 doses after surgery. Thirty-four months after surgery, the cat remains well and has no radiographic evidence of pulmonary metastasis. PMID- 15135358 TI - Spin trapping of nitric oxide by aci anions of nitroalkanes. AB - In alkaline solutions, nitroalkanes (RCH2NO2) undergo deprotonation and rearrange to an aci anion (RHC=NO2-), which may function as a spin trap. Using electron paramagnetic resonance (EPR) spectroscopy, we have investigated suitability of aci anions of a series of nitroalkanes (CH3NO2, CH3CH2NO2, CH3(CH2)2NO2, and CH3(CH2)3NO2) to spin trap nitric oxide (*NO). Based on the observed EPR spectra, the general structure of the adducts, formed by addition of *NO to RHC=NO2-, was identified as nitronitroso dianion radicals of general formula [RC(NO)NO2]*2- in strong base (0.5 M NaOH), and as a mono-anion radical [RCH(NO)NO2]*- in alkaline buffers, pH 10-13. The hyperfine splitting on 14N in the -NO2 moiety (11.2-12.48 G) is distinctly different from the splitting on 14N in the -NO moiety of the adducts (5.23-6.5 G). The structure of the adducts was verified using 15N-labeled *NO, which produced radicals, in which triplet due to splitting on 14N (I = 1) in 14NO/aci nitro adducts was replaced by a doublet due to 15N (I = 1/2) in 15NO/aci nitro adducts. EPR spectra of aci nitromethane/NO adduct recorded in NaOH and NaOD (0.5 M) showed that the hydrogen at alpha-carbon can be exchanged for deuterium, consistent with structures of the adducts being [CH(NO)NO2]*2- and [CD(NO)NO2]*2-, respectively. These results indicate that nitroalkanes could potentially be used as prototypes for development of *NO-specific spin traps suitable for EPR analysis. PMID- 15135359 TI - The mechanisms of S-nitrosothiol decomposition catalyzed by iron. AB - The mechanisms of S-nitrosothiol transformation into paramagnetic dinitrosyl iron complexes (DNICs) with thiol- or non-thiol ligands or mononitrosyl iron complex (MNICs) with N-methyl-D-glucamine dithiocarbamate catalyzed by iron(II) ions under anaerobic conditions were studied by monitoring EPR or optical features of the complexes and S-nitrosothiols. The kinetic investigations demonstrated the appearance of short-living paramagnetic mononitrosyl-iron complex with L-cysteine prior to the formation of stable dinitrosyl-iron complex with cysteine in the solution of iron(II)-citrate complex (50-100 microM), S-nitrosocysteine (400 microM), and L-cysteine (20 mM) in 100 mM Hepes buffer (pH 7.4). The addition of deoxyhemoglobin (100 microM) did not influence the process, which points to a direct interaction between S-nitrosocysteine and iron(II) ions to yield DNIC. The reaction of DNIC-cysteine formation is first- and second-order in iron and S nitrosocysteine, respectively. The third-order rate constant is (1.0 +/- 0.2) x 10(5) M(-2) s(-1) (estimated from EPR results) or (2.0 +/- 0.1) x 10(4) M(-2) s( 1) (estimated by optical method). A similar process of DNIC-cysteine formation was observed in a solution of iron(II)-citrate complex, L-cysteine, and NO proline (200 microM) as a NO* donor. The appearance of a less stable dinitrosyl iron complex with phosphate was detected when solutions of iron(II)-citrate containing 100 mM phosphate buffer (pH 7.4) were mixed with S-nitrosocysteine or NO-proline. The rapid formation of DNIC with phosphate was followed by its decay. When the concentration of L-cysteine in solutions was reduced from 20 to 1 mM, the life-time of the DNIC-cysteine diminished notably; this was caused by consumption of L-cysteine in the process of DNIC-cysteine formation from S nitrosocysteine and iron. Thus, L-cysteine is consumed. Formation of DNIC with glutathione was also observed in a solution of glutathione (20 mM), S nitrosoglutathione (400 microM), and iron(II) complex (800 microM) in 100 mM Hepes buffer (pH 7.4), but the rate of formation was about 10 times slower than the formation of the DNIC-cysteine. The rate of MNIC-MGD formation from iron(II) MGD complexes and S-nitrosocysteine was first-order in both reactants. The second order rate constant for this reaction, estimated from EPR measurements, was 30 +/ 5 M(-1) s(-1). Rate constants of MNIC-MGD formation from iron(II)-MGD and the more stable S-nitrosoglutathione and S-nitroso-D,L-penicillamine were equal to 3.0 +/- 0.3 and 0.3 +/- 0.05 M(-1) s(-1), respectively. Thus, the concerted mechanism of DNIC and MNIC formation from S-nitrosothiols and iron(II) ions can be suggested to be predominant. PMID- 15135360 TI - Reductive nitrosylation and S-nitrosation of hemoglobin in inhomogeneous nitric oxide solutions. AB - Elucidating the reaction of nitric oxide (NO) with oxyhemoglobin [HbFe(II)O2] is critical to understanding the metabolic fate of NO in the vasculature. At low concentrations of NO, methemoglobin [HbFe(III)] is the only detectable product from this reaction; however, locally high concentrations of NO have been demonstrated to result in some iron-nitrosylhemoglobin [HbFe(II)NO] and S nitrosohemoglobin (SNO-Hb) formation. Reductive nitrosylation through a HbFe(III) intermediate was proposed as a viable pathway under such conditions. Here, we explore another potential mechanism based on mixed valenced Hb tetramers. The oxidation of one or two heme Fe(II) in the R-state HbFe(II)O2 has been observed to lower the oxygen affinity of the remaining heme groups, thus creating the possibility of oxygen release and NO binding at the heme Fe(II) sites. This mixed valenced hypothesis requires an allosteric transition of the Hb tetramer. Hence, this hypothesis can account for HbFe(II)NO formation, but not SNO-Hb formation. Here, we demonstrate that cyanide attenuated the formation of SNO-Hb by 30-40% when a saturated NO bolus was added to 0.1-1.0 mM HbFe(II)O2 solutions. In addition, HbFe(II)NO formation under such inhomogeneous conditions does not require allostericity. Therefore, we concluded that the mixed valenced theory does not play a major role under these conditions, and reductive nitrosylation accounts for a significant fraction of the HbFe(II)NO formed and approximately 30 40% of SNO-Hb. The remaining SNO-Hb is likely formed from NO oxidation products. PMID- 15135361 TI - A macrocyclic nitrosyl ruthenium complex is a NO donor that induces rat aorta relaxation. AB - The vasorelaxation induced by a nitrosyl macrocyclic ruthenium complex, proposed as a new nitric oxide (NO) carrier, was studied in rat isolated aorta. The compound trans-[RuCl([15]aneN4)NO]2+ was characterized by elemental analysis, UV visible spectrum, and infrared spectrum. Based on the electrochemical process, the reduction of the compound was followed by NO release, which was also observed using norepinephrine as a reducing agent and NO released was analyzed by a sensor. Vasorelaxation induced by this NO donor was studied and compared to those obtained with sodium nitroprusside (SNP). The relaxation induced by the compound was concentration-dependent in denuded rat aortas and occurred only in pre contracted arteries with norepinephrine. The macrocyclic compound induced relaxation with a similar efficacy as SNP, although the potency of SNP was slightly greater. The time to reach maximum relaxation (595 s) was longer than that of SNP (195 s). Relaxation was completely abolished by oxyhemoglobin, a known NO scavenger. PMID- 15135362 TI - Interleukin-1beta up-regulates nitrite production: effects on ovarian function. AB - We have previously reported that Interleukin-1beta (IL-1beta) affects ovarian function in the rat, modulating prostaglandin and progesterone (P) production. As IL-1beta effects were associated to nitric oxide (NO) synthesis, in the present work we have further examined the role of ovarian NOS-system, in IL-1beta antisteroidogenic action. Mid-luteal explants from rats were incubated for 4 h in the presence of IL-1beta (1-35 ng/ml)-alone or in combination with NOS-inhibitors and then assayed for P and nitrite production. IL-1beta treatment reduced P levels in a dose-dependent manner, returning to basal levels at 35 ng/ml. This reduction in steroid synthesis was paralleled by a dose-dependent increase in nitrite levels, reaching a maximum at 25 ng/ml but without effect at 35 ng/ml. L Arginine (1 and 2 mM) was able to mimic IL-1beta actions and the NOS blocker L Nitro-Arginin-Methyl Ester reverted these effects. Moreover, the selective iNOS inhibitor, 1400 W, completely abolished IL-1beta antisteroidogenic effect, therefore confirming the dependence of IL-1beta action upon iNOS activation. Finally, IL-1beta did not affect eNOS expression but up-regulated iNOS mRNA and protein levels. Our results suggest an interaction between IL-1beta and the NOS system. Thus, we may conclude that in the rat iNOS-derived NO production, induced by IL-1beta, affects ovarian P biosynthesis and hence NO may be a major effector molecule of ovarian IL-1 system. PMID- 15135364 TI - Analysis of cytokine-induced NO-dependent apoptosis using RNA interference or inhibition by 1400W. AB - RNA interference has been used to silence gene expression and evaluate the contribution of a gene product to cell function. Here, we investigated conditions under which expression of an inducible protein, nitric oxide synthase 2 (NOS2), is decreased by RNA interference. Cytokine treatment of insulin-producing RINm5F cells results in NOS2 induction and cell death. Conditions used here favoured cytokine-induced apoptosis, for the first time--rather than necrosis, previously recorded. In RINm5F cells, transfected with NOS2-specific small interfering RNA followed by a 12 h exposure to cytokines, there was a significant reduction in NOS2 protein, nitrite, and apoptosis. There were no changes in these three parameters when experiments were carried out with unrelated vimentin siRNA. To interpret the NOS2-siRNA result further, we compared it with complete pharmacological inhibition of nitric oxide (NO) production by the NOS2 competitive inhibitor, 1400W, which lowered apoptosis by only 50% in the RINm5F cells. We conclude that the use of NOS2-specific siRNA has resulted in the subsequent lowering of expression of a cytokine-inducible protein whose function can be quantified. siRNA results have compared favourably with use of a pharmacological inhibitor of NOS2, in revealing the subtle, partial contribution of cytokine-induced NO to apoptosis induction in these insulin-producing cells. PMID- 15135363 TI - Neuronal NOS deficiency promotes apoptotic cell death of spinal cord neurons after peripheral nerve transection. AB - To study the role of endogenous NO in survival and recovery of spinal cord neurons after nerve lesions, wild type mice were compared to knock-out mice lacking neuronal, endothelial or inducible NO synthase (NOS) after sciatic nerve transection. The NO-generating capacities were assessed by NOS immunohistochemistry and NADPH-diaphorase staining. The feature of affected neurons was evaluated following Nissl- and TUNEL-staining, by immunocytochemical demonstration of cytochrome c-translocation, and by ultrastructural examination. Time point of cell loss was found to be independent of the mice type and occurred only at later post-axotomy states. The extent of neuronal degeneration, however, depended on the NO supply. Whereas a lack of endothelial or inducible NOS was well tolerated, deficiency of neuronal NOS enhanced the competence-to-die and led to a substantial apoptotic cell death of spinal cord neurons. Thus, NO supply turned out to be essential for cell survival and recovery with reference to the neuronal NOS isoform. PMID- 15135365 TI - Potential applications and prospects for cardiac xenotransplantation. AB - Despite improvements in pharmacologic therapies, the outlook for patients with severe cardiac disease remains poor. At present, the only "cure" for end-stage heart failure is transplantation. However, fewer than 5% of those who need a cardiac transplant receive one in the United States each year. As an alternative, some propose using animals as a source of organs for transplantation (i.e., xenotransplantation). In this article we review the potential applications of xenotransplantation for the treatment of cardiac disease, and weigh xenotransplantation against other new technologies that might be used. We also consider the current status of addressing the hurdles to application of xenotransplantation. PMID- 15135366 TI - Microvascular changes in small airways predispose to obliterative bronchiolitis after lung transplantation. AB - BACKGROUND: There is strong evidence that obliterative bronchiolitis (OB) in lung transplant recipients is related to acute rejection as graded by parenchymal perivascular infiltrates. OB (chronic rejection) is a small airways, rather than a parenchymal, scarring process. Moreover, there has been no study of the microcirculation in the small airways in lung transplantation. This study assesses the microvasculature around small airways (SA) in post-mortem lung allograft specimens. METHODS: The microvasculature of SA (n = 19) from 5 patients who died within 24 hours of lung transplantation (Group A) and SA in OB lungs (11 patients, median post-transplant survival 1,371 days) was assessed by the use of monoclonal antibodies to the vascular endothelium, namely von Willebrand factor (vWF) and CD31. The second group was further sub-divided into Group B (airways not obliterated, n = 18), Group C (sub-total airways obliteration, n = 21) and Group D (airways with total luminal obstruction, n = 14). RESULTS: The measured median circumference of the SA in the 4 groups was 2.1, 2.1, 2.5 and 2.3 mm, respectively (p = 0.66). Using CD31 as the endothelial marker, the median number of blood vessels per unit length of airway circumference (BVPL) was 3.5 vessels/mm for Group A, 0.8 for Group B, 1.3 for Group C and 2.8 for Group D, (p < 0.001). Large blood vessels (circumference >0.20 mm) were present in 95%, 11%, 14% and 21% of each group, respectively (p < 0.001). Similar trends were confirmed with the vWF endothelial antibodies. CONCLUSIONS: OB after lung transplantation is associated with a decrease in microvascular supply to the small airway. This ischemic event may lead to airway damage or increase the tendency to repair by scarring. The small airways then appear to respond to this insult by angiogenesis, which may either occur too late to prevent permanent airway damage or be inadequate in restoring adequate blood supply to the airway. PMID- 15135367 TI - Bronchoalveolar lavage fluid characteristics in acute and chronic lung transplant rejection. AB - BACKGROUND: The detection of graft rejection by bronchoalveolar lavage remains controversial. METHODS: To assess the value of bronchoalveolar lavage fluid in acute and chronic rejection after lung transplantation we analyzed bronchoalveolar lavage fluid cellular differential characteristics, lymphocyte sub-types and interleukin-6 (IL-6) and interleukin-8 (IL-8) cytokine levels in patients with exclusively either acute rejection (n = 37) or bronchiolitis obliterans (BO; n = 48). Both groups were compared with a control group of lung transplantation patients without rejection or infection, matched for the time the lavage was performed after lung transplantation. RESULTS: The bronchiolitis obliterans group showed marked neutrophilia, high IL-8 and higher CD4(+)CD25(+) and CD8(+)CD45(+) bronchoalveolar lavage fluid levels when compared with their stable controls. When using a cut-off point of >3% neutrophils in the lavage, the sensitivity for BO is 87.0%, the specificity 77.6%. The sensitivity of IL-8 for BO when using a cut-off point of >71.4 pg/ml is 74.5%, the specificity 83.3%. Bronchoalveolar lavage fluid in acute rejection was characterized by marked lymphocytosis, but showed no difference when compared with stable controls in any of the lymphocyte sub-types studied. When using a cut-off point of <==1% lymphocytes in the lavage, the sensitivity for acute rejection (AR) is 40.4%, the specificity 95.6%. The marked neutrophilia, high IL-8 cytokine level and more activated lymphocyte population in bronchiolitis obliterans may indicate ongoing local allograft rejection. CONCLUSIONS: In the present study we were not able to show any difference in lymphocyte sub-types when comparing acute rejection and control subjects. Cellular and soluble parameters in bronchoalveolar lavage fluid appear useful for diagnosing bronchiolitis obliterans. PMID- 15135368 TI - Interleukin-10 production genotype protects against acute persistent rejection after lung transplantation. AB - BACKGROUND: Our previous studies demonstrated that cytokine gene polymorphisms are related to acute rejection in pediatric heart transplantation; a decreased tumor necrosis factor (TNF)-alpha production genotype combined with an increased or intermediate interleukin (IL)-10 production genotype was associated with the smallest incidence of acute rejection. The objective of this study was to determine whether cytokine genotypes TNF-alpha, IL-10, IL-6, interferon-gamma, and transforming growth factor beta were associated with acute persistent rejection after lung transplantation. METHODS: Cytokine genotyping was performed in 119 adult lung transplantation recipients who underwent surveillance transbronchial biopsies during their first year after transplantation. We categorized recipients with acute persistent rejection if they had 2 consecutive biopsy specimens at >/=Grade A2 despite anti-rejection treatment. We performed cytokine genotyping using the polymerase chain reaction-sequence specific primers technique, with a commercially available kit. RESULTS: We analyzed the IL-10 genotype in 116 patients. For the increased IL-10 production genotype, 7 of 20 patients (35%) were persistent rejecters. In comparison, 57 of 96 patients (59%) with intermediate or decreased IL-10 production genotype had acute persistent rejection (p = 0.046). For IL-10 haplotypes associated with intermediate IL-10 production, 30 of 45 patients with GCC/ACC haplotype (67%) had acute persistent rejection compared with 10 of 22 patients with GCC/ATA (45%). In the patients with intermediate IL-10 production, 17 of 22 (77%) with IL-10 GCC/ACC and IL-6 G/C had acute persistent rejection, whereas only 2 of 7 patients (29%) with IL-10 GCC/ATA and IL-6 G/G had acute persistent rejection (p = 0.018). CONCLUSIONS: In lung transplant recipients, the increased IL-10 production genotype protects against acute persistent rejection when compared with the intermediate or decreased IL-10 production genotypes. The intermediate IL-10 production genotype in lung transplant recipients can be differentiated into 2 haplotype responses, with the GCC/ACC haplotype associated more with acute persistent rejection. In lung transplant recipients, the immunomodulatory effects of IL-6 are differentiated in the G/C and G/G alleles in conjunction with IL-10 haplotypes, with G/C being associated with more acute persistent rejection in conjunction with the IL-10 GCC/ACC haplotype. Future pharmacogenomic models may incorporate these associations with acute persistent rejection in lung transplant recipients to formulate individualized therapeutic regimens. PMID- 15135369 TI - How to predict forced vital capacity after living-donor lobar-lung transplantation. AB - BACKGROUND: Living-donor lobar-lung transplantation (LDLLT) has evolved from a rarely performed experimental procedure to an accepted therapy for selected patients who are unlikely to survive the long wait for cadaveric lungs. However, a convincing study has not been performed that shows the effects of small grafts and of pre-operative variables in predicting functional outcome of recipients after LDLLT. METHODS: From October 1998 to March 2002, 2 male and 11 female patients underwent LDLLT. Mean age was 27.3 years (range, 8-53 years). Diagnoses included primary pulmonary hypertension (n = 5), idiopathic interstitial pneumonia (n = 2), bronchiolitis obliterans (n = 2), bronchiectasis (n = 2), lymphangioleiomyomatosis (n = 1), and cystic fibrosis (n = 1). Donors included 12 men and 14 women with a mean age of 40 years. Given that the right lower lobe consists of 5 segments, the left lower lobe of 4, and the whole lung of 19, we estimated the graft forced vital capacity (FVC) based on the donor's measured FVC and compared this with the recipient's FVC measured after LDLLT. RESULTS: Currently, all patients are alive, with a mean follow-up of 22.2 months (range, 10-51 months). The recipients' FVC measured at 6 months (1,813 +/- 86 ml) correlated well with the graft FVC (1,803 +/- 70 ml), estimated based on the donors' measured FVC (r = 0.802, p = 0.00098). CONCLUSIONS: Recipient FVC after LDLLT can be predicted by measuring donor FVC before surgery regardless of the diagnosis of the recipient. PMID- 15135370 TI - Non-ischemic left ventricular dysfunction after pediatric cardiac transplantation: treatment with plasmapheresis and OKT3. AB - BACKGROUND: Acquired left ventricular dysfunction after pediatric cardiac transplantation is associated with a high mortality rate. It often occurs without biopsy evidence of cellular rejection or severe transplant coronary arteriopathy. METHODS: We employed a protocol for treatment of acquired, non-ischemic left ventricular dysfunction utilizing plasmapheresis, monoclonal anti-T-cell antibody (OKT3), cyclophosphamide and steroids, regardless of the results of endomyocardial biopsy. Left ventricular dysfunction was defined as an echocardiographic shortening fraction of <29% and/or symptoms of congestive heart failure requiring inotropic support. Transplant coronary arteriopathy was excluded by coronary angiography in all cases. RESULTS: Ten pediatric heart transplant recipients were treated for 13 episodes of non-ischemic left ventricular dysfunction. Biopsy scores were low grade (ISHLT Grade 1A or 1B) in 8 episodes. Eight of 10 patients had a history of non-compliance in regularly taking immunosuppressant medications. Inotropic support was required in 9 of 13 cases, with a median duration of 5 days. Median left ventricular shortening fraction was 17% at time of presentation. Normalization of shortening fraction occurred a median of 40 days from the start of treatment. Survival to hospital discharge occurred in 11 of 13 (85%) patients. Long-term patient survival, however, was only 50% at 24 months after presentation with a first episode of acquired left ventricular dysfunction. CONCLUSIONS: Use of plasmapheresis, OKT3, cyclophosphamide and steroids resulted in successful short-term reversal of non ischemic left ventricular dysfunction in pediatric heart transplant patients, but long-term survival remained poor. PMID- 15135371 TI - B-type natriuretic peptide in children after cardiac transplantation. AB - BACKGROUND: The normal B-type natriuretic peptide (BNP) concentration and the significance of increased BNP concentration in children after orthotopic heart transplant (OHT) remain unknown. We sought to determine plasma BNP concentrations in relation to post-transplant time and to explore factors associated with increased BNP in pediatric OHT recipients. METHODS: We obtained plasma BNP concentrations in 44 pediatric patients at 1 to 171 months after OHT. All patients underwent endomyocardial biopsies and echocardiography. We analyzed the association between BNP and post-transplant time, as well as the association between BNP concentration and left ventricular end-diastolic dimension (LVEDD) after transplantation. RESULTS: The age of study patients ranged from 0.9 to 21.2 years (mean, 11.4 +/- 6.2 years; median, 10.5 years). We evaluated BNP concentrations immediately after transplantation. The mean BNP concentration decreased exponentially to 100 pg/ml by 14 weeks after OHT. Although BNP concentration relative to time after OHT varied among individuals, all patients with multiple measurements showed predictable rates of decrease. This decrease in BNP concentration was not associated with changes in LVEDD. CONCLUSIONS: Plasma BNP concentration was elevated in children after OHT and decreased exponentially in time to 100 pg/ml by 14 weeks after OHT. The decrease in BNP concentration was unrelated to left ventricular dimension. PMID- 15135372 TI - Management of aortopulmonary collaterals in children following cardiac transplantation for complex congenital heart disease. AB - BACKGROUND: Heart transplantation (HTx) is increasingly utilized as therapy for end-stage cyanotic congenital heart disease. This study investigates the presence and impact of aortopulmonary collaterals (APCs) associated with cyanotic heart disease on the early post-operative course of patients undergoing transplantation. High output cardiac failure due to residual aortopulmonary collaterals can affect outcome following heart transplantation. METHODS: Seven patients with hemodynamically significant APCs post-transplant were identified among 40 patients with cyanotic congenital heart disease undergoing HTx. The peri and intra-operative courses of these patients were reviewed. All 7 patients required prolonged inotropic support despite normal ventricular function and no allograft rejection; 5 were ventilator-dependent due to significant pulmonary vascular congestion. Selective angiography demonstrated the presence of significant aortopulmonary collaterals at 7 to 19 days post-transplant. Coil embolization of aortopulmonary collaterals was performed in all patients; a mean of 6 (2 to 16) vessels/patient were embolized. RESULTS: After embolization, pulmonary edema resolved and heart size normalized in all patients; inotropic support was weaned within 2 to 10 days in 5 patients. One patient developed transient renal failure secondary to excessive contrast load and another had enterococcal sepsis within 24 hours after the procedure. All patients were asymptomatic from 4 to 10 years of follow-up post-HTx. CONCLUSIONS: Aortopulmonary collaterals should be considered a cause of early donor heart failure in children following HTx for cyanotic congenital heart disease. Early detection and treatment of aortopulmonary collaterals by coil embolization is necessary to improve the post-transplant course in these complex patients. PMID- 15135374 TI - Bone marrow is a source of regenerated cardiomyocytes in doxorubicin-induced cardiomyopathy and granulocyte colony-stimulating factor enhances migration of bone marrow cells and attenuates cardiotoxicity of doxorubicin under electron microscopy. AB - BACKGROUND: It has been reported previously that granulocyte colony-stimulating factor (GCSF) injection improves infarcted heart function, but the mechanism remains unclear. In this study we sought to determine whether GCSF-mobilized bone marrow cells could regenerate neo-myocardium and repair doxorubicin-induced cardiomyopathy. METHODS: C57BL/6 mice were irradiated and bone marrow cells (BMC; 1 x 10(6)) from green fluorescent protein (GFP) mice (GFP-BMC) were transplanted intravenously, followed by splenectomy. Doxorubicin (2.5 mg/kg, 6 times for 2 weeks) was administered intraperitoneally 2 weeks later. GCSF (50 microg/kg/day for 8 days) was administered sub-cutaneously after doxorubicin injection (Group I, n = 11) and 3 weeks later (Group II, n = 8), and saline was injected in Group III animals (n = 8). Eight weeks after doxorubicin injection, the excised hearts were studied immunologically and electron microscopically. RESULTS: Survival rates were 81.8% in Group I, 50.0% in Group II and 62.5% in Group III. The number of GFP-BMC in Group I (15.4 +/- 7.4 per high-power field) was highest (p < 0.05). In all groups, cardiac troponin I-positive cells derived from GFP-BMC were observed in the hearts. GFP-BMC in hearts stained positively against cardiac troponin I (4.3 +/- 2.5%), myosin heavy chain (5.0 +/- 4.3%), atrial natriuretic peptide (ANP; 3.9 +/- 2.4%) and connexin 43 (11.9 +/- 7.3%) in Group I. Myofibrils, mitochondria and fundamental architecture were almost all preserved in Group I, whereas hearts were severely damaged in Groups II and III. CONCLUSIONS: Bone marrow was shown to be one of the sources of regenerated cardiomyocytes in the doxorubicin-induced cardiomyopathic heart. Early administration of GCSF enhanced the migration of bone marrow cells into the heart, and attenuated the cardiotoxicity of doxorubicin. PMID- 15135373 TI - Clinical usefulness of bronchoalveolar lavage in heart transplant recipients with suspected lower respiratory tract infection. AB - BACKGROUND: Bronchoscopy with bronchoalveolar lavage (BAL) is the recommended initial invasive diagnostic procedure when lower respiratory tract infection is suspected in solid-organ transplant recipients. In this study, we evaluated the clinical impact and safety of bronchoscopy with BAL in heart transplant recipients. METHODS: We reviewed all 44 consecutive diagnostic bronchoscopies with BAL that were performed in 35 heart transplant recipients at Helsinki University Central Hospital between May 1988 and December 2001. RESULTS: Bronchoscopy findings established specific microbiologic diagnoses in 18 of 44 (41%) cases, and 14 of 44 (32%) bronchoscopic findings led to changes in therapy. The diagnostic yield of bronchoscopy from 1 to 6 months after transplantation was 73%, significantly better (p = 0.002) than diagnostic yield during the first month (18%) and after 6 months (28%). Pneumocystis carinii and cytomegalovirus were the most frequently detected pathogens in the BAL fluid. Cytomegalovirus pneumonia carried a high mortality rate (44%), whereas all patients with P carinii pneumonia recovered. Fourteen episodes were diagnosed as bacterial pneumonia, but because of empiric antibiotic therapy that was started widely before bronchoscopy, a microbiologic diagnosis was established in only 1 case. However, all patients with community-acquired pneumonia responded to empiric therapy. Four cases of major complications occurred after bronchoscopy, all cardiovascular but none fatal. CONCLUSIONS: Bronchoscopy with BAL is a useful diagnostic tool in heart transplant recipients, especially between 1 and 6 months after transplantation. PMID- 15135375 TI - Alpha chemokines regulate direct lung ischemia-reperfusion injury. AB - BACKGROUND: Alpha chemokines function predominantly to recruit and activate neutrophils, which are important effectors of acute lung injury. This study evaluated whether blockade of 2 potent alpha chemokines, macrophage inflammatory protein-2 (MIP-2) and cytokine-induced neutrophil chemoattractant (CINC), is protective against lung ischemia-reperfusion injury in a warm in situ hilar clamp model. METHODS: Left lungs of Long-Evans rats underwent normothermic ischemia for 90 minutes and reperfusion for up to 4 hours. Treated animals received antibodies to MIP-2 or CINC immediately prior to reperfusion. Lung injury was quantitated by vascular permeability to (125)I-radiolabeled bovine serum albumin, lung tissue neutrophil sequestration (myeloperoxidase [MPO] content), and alveolar leukocyte content in bronchoalveolar lavage (BAL) fluid. CINC and MIP-2 mRNA expression were assessed by northern blot, while ribonuclease protection assays were performed to evaluate mRNA expression for a number of early response cytokines. MIP-2 and CINC protein expression in injured lungs was determined by immunoblotting. RESULTS: Treatment with antibodies to CINC or MIP-2 was associated with significant protection against increases in vascular permeability, MPO content and alveolar leukocyte sequestration in injured lungs. Expression of CINC and MIP-2 mRNA peaked after 2 hours of reperfusion in injured lungs, and protein levels were evident on immunoblotting after 3 hours of reperfusion. Neither CINC nor MIP-2 blockade appeared to modulate cytokine mRNA expression. CONCLUSIONS: CINC and MIP-2 are important mediators involved in direct lung ischemia-reperfusion injury. They appear to function by modulating neutrophil recruitment, but not inflammatory cytokine release. PMID- 15135376 TI - Endothelium-dependent relaxation of canine pulmonary artery after prolonged lung graft preservation in University of Wisconsin solution: role of L-arginine supplementation. AB - BACKGROUND: The University of Wisconsin (UW) solution has been demonstrated to enhance pulmonary allograft preservation. Endothelial nitric oxide (NO) production has been shown to be significantly impaired after ischemia and reperfusion (I/R) injury. The present experiments aimed to determine the protective effects of pulmonary endothelium-dependent function by using supplemental NO in University of Wisconsin (UW) solution following prolonged lung graft preservation. METHODS: Thirty-six healthy mongrel dogs underwent thoracotomy to expose the left lung. In addition to a group given UW solution (n = 4), 100 micromol/liter l-arginine, (n = 7), 100 micromol/liter N(G)-monomethyl l-arginine (l-NMMA n = 7) and 1.0 micromol/liter 3-morpholinosydnonimine (SIN-1, n = 18 respectively, were added to UW solution, and infused from the aortic root and pulmonary artery to the pulmonary vein. The perfused lung was then allowed to inflate to its maximum volume for 24-hour oxygenated preservation in each supplemented condition of UW solution at 4 degrees C. In the SIN-1 group, the preservation period was further divided into 8 hours and 16 hours, respectively. Rings of the third-order pulmonary artery of the inflated lung were then suspended in organ chambers to measure isometric force. RESULTS: Endothelium dependent relaxation (EDR) to acetylcholine, adenosine diphosphate and sodium fluoride of the pulmonary rings in the l-arginine group was significantly preserved compared with UW-solution-only group. The l-NMMA group showed significant EDR impairment after 24-hour preservation compared with the UW solution group. Similar to the l-arginine group, the SIN-1 group showed significant EDR protection with 8-hour preservation, but not with 24-hour preservation. In contrast, EDR to calcium ionophore A23187 showed no EDR changes after 24-hour preservation in any of the supplemented groups. CONCLUSIONS: Supplemental l-arginine in UW solution ameliorates impaired pulmonary EDR following prolonged lung preservation of up to 24 hours. PMID- 15135377 TI - Heme oxygenase does not contribute to control of basal vascular tone in isolated blood-perfused rat lung. AB - BACKGROUND: Vasoconstriction in pulmonary ischemia-reperfusion injury may involve dysfunction of the physiologic vasodilation of pulmonary arteries. Little is known of the relative importance of heme oxygenase (HO)/carbon monoxide (CO) dependent vs nitric oxide synthase (NOS)/nitric oxide (NO)-dependent vasodilation of the pulmonary vasculature. We evaluated the significance of HO function on basal pulmonary vascular resistance (PVR) and compared it with the function of NOS. METHODS: Using an isolated blood-perfusion model, lungs of Lewis rats were assigned to 3 groups (n = 6/group). After stabilization, either an inhibitor of HO (tin-protoporphyrin-9 [SnPP-9]) or an inhibitor of NOS (NG-nitro-L-arginine methylester [L-NAME]) was added to the perfusate (50 micromol/liter and 1 mmol/liter as the final concentration, respectively). Lungs receiving saline served as controls. Gas exchange, hemodynamic and respiratory functions and the levels of cyclic 3',5'-guanosine monophosphate (cGMP) in the perfusate were measured. RESULTS: Inhibition of NOS by L-NAME resulted in a significant (p < 0.01) increase in PVR (DeltaPVR: 0.110 +/- 0.012 cm H(2)O/ml. min) within 5 minutes. In contrast, PVR was minimally affected by SnPP-9 (DeltaPVR: 0.005 +/- 0.005 cm H(2)O/ml. min), which was comparable to control lungs (DeltaPVR: 0.012 +/- 0.005 cm H(2)O/ml. min). The level of cGMP in the perfusate 5 minutes after drug application was markedly, but not significantly, lower in the L-NAME group (1.67 +/- 0.74 nmol/liter) when compared with controls (2.69 +/- 0.89 nmol/liter) and SnPP-9-treated lungs (2.65 +/- 0.66 nmol/liter). CONCLUSIONS: NOS but not HO contributes to the control of basal vascular tone in the rat lung. PMID- 15135379 TI - Physical and psychological attributes of fatigue in female heart transplant recipients. AB - BACKGROUND: The attributes of fatigue after orthotopic heart transplantation (OHT) are poorly understood. We conducted this study to assess the prevalence, severity and correlates of fatigue among female OHT survivors. METHODS: Fifty women (age 54.7 +/- 13.0 years) from a single heart transplant center, who underwent OHT 5.1 +/- 4.4 (mean +/- SD) years earlier, completed a battery of questionnaires including the Profile of Mood States-fatigue sub-scale to assess levels of fatigue, the Beck Depression Inventory to measure depression, and the Short Form-36 to measure functional status and mental health. Demographic and clinical data were obtained from self-reports and medical chart reviews. RESULTS: Women reported mean scores of 15.7 +/- 6.8 (range 3 to 27), 13.2 +/- 8.2 (range 0 to 38), 37.2 +/- 10.8 (range 22 to 62) and 41.5 +/- 11.2 (range 17 to 60) for fatigue, depression, functional status and mental health, respectively. Univariate analyses revealed that sociodemographic and clinical variables (e.g., age, employment status, anemia, renal insufficiency) were significantly related to fatigue (p < 0.001). Likewise, depression, functional status and mental health were also significantly related to fatigue (p < 0.001). In a multivariate model, age (adjusted R(2) = 0.23, p < 0.001), anemia (adjusted R(2) = 0.39, p < 0.001), functional status (adjusted R(2) = 0.60, p < 0.001) and depression (adjusted R(2) = 0.69, p < 0.001) were significant predictors of fatigue. The model explained 69% of the variance in fatigue (p < 0.001). CONCLUSIONS: Fatigue is common in women after OHT and is associated with both physiologic and psychologic factors. Clinicians should evaluate all female recipients for symptoms of fatigue, especially those with anemia, renal insufficiency, poor functional status and depression. Other potential mediators of fatigue, such as the denervated donor heart and type of immunosuppressive regimen, may also play a role and require further study. PMID- 15135378 TI - Effects of a bradykinin B(2) receptor antagonist on ischemia-reperfusion injury in a canine lung transplantation model. AB - BACKGROUND: This study investigated the effects of a bradykinin B(2) receptor antagonist, FR173657 (FR), on ischemia-reperfusion (I/R) injury in a canine lung transplantation model. METHODS: Eighteen pairs of weight-matched dogs were randomly divided into 3 groups. Six pairs were assigned to the FR(D+R) group, in which FR (100 nmol/kg/h) was administered to the transplant donor continuously beginning 30 minutes before ischemia until the onset of ischemia, and FR was administered to the transplant recipient beginning 30 minutes before reperfusion and continuing for 2 hours after reperfusion. Another 6 pairs of dogs were assigned to the FR(R) group, in which FR was administered only to the recipient in the same manner as in the FR(D+R) group. The other pairs were assigned to the control group, in which vehicle alone was administered. Orthotopic left lung transplantation was performed after 12-hour cold storage in Euro-Collins solution. Fifteen minutes after reperfusion, the right pulmonary artery and the right stem bronchus were ligated. The animals were measured for 4 hours after reperfusion for left pulmonary vascular resistance (L-PVR), cardiac output (CO), arterial oxygen pressure (PaO(2)) and alveolar-arterial oxygen pressure difference (A-aD(O(2))). Lung specimens were harvested for measurement of the wet to-dry lung weight ratio (WDR), histopathologic studies and polymorphonuclear neutrophil (PMN) count. RESULTS: Compared with the control group, PaO(2), A aDO(2), L-PVR and CO were all significantly (p < 0.05) improved and WDR significantly (p < 0.05) lower in both the FR(D+R) and FR(R) groups. Moreover, in the FR-treated groups, histologic tissue edema was mild, and PMN infiltration was significantly (p < 0.05) reduced. CONCLUSIONS: The bradykinin B(2) receptor antagonist, FR173657, ameliorates I/R injury in lung grafts, indicating that protection of lung grafts can be achieved by the administration of FR solely to the transplant recipient. PMID- 15135380 TI - Effects of changes in UNOS policy regarding left ventricular assist devices. AB - Over the last 6 years, 161 patients underwent implantation of HeartMate single lead vented electric (SLVE) devices as a bridge to transplantation. SLVE patients were retrospectively analyzed before and after United Network for Organ Sharing (UNOS) policy changes, and designated SLVE-1 and SLVE-2, respectively. Mean support time decreased significantly from 79.7 +/- 72.0 days to 55.3 +/- 60.5 days (p = 0.022). Although not statistically significant, rate of successful bridging to transplant increased from 70.3% to 74.7%, and post-transplant 1- and 3-year survival increased from 90.0% and 86.0%, respectively, to 92.9% and 87.3%, respectively. PMID- 15135381 TI - Non-specific removal of antibodies in patients with idiopathic dilated cardiomyopathy: implications for xenotransplantation. AB - We assessed the efficiency of non-specific extracorporeal immunoadsorption (EIA), using polyclonal anti-human immunoglobulin antibodies, in depleting the serum of anti-galactosealpha1,3galactose (Gal) antibody and in decreasing serum cytotoxicity in 5 patients with idiopathic dilated cardiomyopathy. The mean concentrations of anti-Gal immunoglobulin (Ig)M and IgG before EIA were 74 microg/ml and 159 microg/ml, respectively. After EIA, these concentrations decreased by 86% and 88%, respectively. Both anti-Gal IgM and IgG returned to pre EIA concentrations within 1 month, without rebound to greater than baseline concentrations. After EIA, mean serum cytotoxicity also decreased from 90% to 17%, with recovery by 1 month. Extracorporeal immunoadsorption proved safe in patients with heart failure and was effective in depleting anti-Gal antibody and in decreasing serum cytotoxicity to pig cells. PMID- 15135382 TI - Chylothorax after heart/lung transplantation. AB - Chylothorax is a potentially serious complication of lung and heart-lung transplantation. This article describes the clinical course of chylothorax in 3 heart-lung allograft recipients. We discuss management options, including dietary modifications, octreotide infusion, thoracic duct ligation and embolization, and surgical pleurodesis. In addition, we describe the novel use of aminocaproic acid to reduce lymph flow. We propose a multidisciplinary approach for the management of chylothorax that includes both medical and surgical options. PMID- 15135383 TI - Airway anastomosis complications in de novo lung transplantation with sirolimus based immunosuppression. AB - A prospective, pilot trial was started to evaluate the effect of a sirolimus based immunosuppressive regimen on acute and chronic rejection in de novo lung transplant patients. Primary lung transplant (LTx) recipients received a sirolimus- and tacrolimus-based immunosuppressive therapy immediately after transplantation. Both immunosuppressants were administered with trough level adjusted, while steroid administration was minimized. Four patients were enrolled (2 single-lung transplants, 1 double-lung transplant, 1 heart-lung transplant) in the study. Mean ischemia time was 387 +/- 92 minutes. Acute rejection (at least Grade A1 ISHLT) was detected in 1 patient. Incidence of infection was 0.6 infection per 100 patient-days (3 Aspergillus infections). Until hospital discharge mean sirolimus trough level was 6.2 +/- 1.2 ng/ml. Depending upon mean sirolimus trough levels of each patient, severe wound-healing complications were seen in 3 patients, resulting in bronchial airway dehiscence in 2 patients with lethal outcome in 1 patient. As a result of these complications, we revised the study design after inclusion of only 4 patients: Sirolimus administration is now started after completion of bronchial wound-healing. Sirolimus-based immunosuppressive therapy administered immediately after lung transplantation seems to be associated with severe wound-healing complications of the bronchial anastomosis. PMID- 15135384 TI - Expanding the donor pool: use of a donor heart having undergone recent cardiac surgery. AB - The current shortage of donor organs is a well-recognized global phenomenon. The goal of contemporary transplant practice is to optimize and expand the organ donor pool. Despite biologic and technologic advancements, the single most important limitation remains underutilization and non-recovery of potential organs. We report the use of a donor cardiac organ from a 5-year-old child who had undergone recent (<72 hours) closure of an atrial septal defect. The heart was harvested successfully and implanted with no apparent hemodynamic compromise. PMID- 15135385 TI - Heart transplantation in a patient with multiple sclerosis and mitoxantrone induced cardiomyopathy. AB - We describe a 30-year-old man with end-stage heart failure after therapy with mitoxantrone for multiple sclerosis. A successful orthotopic heart transplantation was performed when intensified medical therapy failed to improve the patient's hemodynamics. In spite of the severe underlying disease he did well on dual immunosuppression with methylprednisone and cyclosporine. Neurologic symptoms remained stable throughout the procedure and, after 2 months, he resumed preoperative ambulatory status. Eight years after the operation, the patient is now in New York Heart Association (NYHA) Class I status. Using canes, he is able to walk short distances. Repeated urinary tract infections caused by Escherichia coli became a problem, but have been controlled by long-term oral antibiotic prophylaxis with trimethoprim. PMID- 15135386 TI - Anastomosis of an anomalous segmental vein with the azygos vein in living-donor lobar lung transplantation. AB - We report the case of a living-donor lobar lung transplantation in which we found an abnormal segmental vein in the right lower lobe of the donor lung. The abnormal vein was anastomosed to the azygos vein of the recipient so that lung edema and congestion could be avoided. This alternative technique is effective in living-donor lobar lung transplantation when the superior segmental vein in the right lower lobe of the donor drains into the superior vein. PMID- 15135387 TI - Pseudoallescheria boydii pneumonia and empyema: a rare complication of heart transplantation cured with voriconazole. AB - Pseudoallescheria boydii pneumonia is a rare occurrence, usually resistant to amphotericin B and other anti-fungal agents. We report a complete response to voriconazole in an immunosuppressed host. PMID- 15135388 TI - Improvement of apparently fixed pulmonary hypertension with cardiac resynchronization therapy. AB - Many patients with advanced heart failure are ineligible for cardiac transplantation because of fixed pulmonary arterial hypertension. Cardiac resynchronization therapy, by stimulating the right atrium, and right and left ventricles, is a new therapy that effectively palliates symptoms in patients with heart failure. Cardiac resynchronization therapy increases cardiac output and decreases pulmonary capillary wedge pressure, thus partially reversing hemodynamic abnormalities that lead to secondary pulmonary hypertension in many heart failure patients. We describe a patient whose previously fixed pulmonary hypertension improved to the point that she was once again considered eligible for cardiac transplantation. PMID- 15135389 TI - ABIOMED's BVS 5000 biventricular support system. PMID- 15135390 TI - Cloning, expression, purification, and characterization of Nocardia sp. GTP cyclohydrolase I. AB - The sequence of the gene from Nocardia sp. NRRL 5646 encoding GTP cyclohydrolase I (GCH), gch, and its adjacent regions was determined. The open reading frame of Nocardia gch contains 684 nucleotides, and the deduced amino acid sequence represents a protein of 227 amino acid residues with a calculated molecular mass of 24,563Da. The uncommon start codon TTG was identified by matching the N terminal amino acid sequence of purified Nocardia GCH with the deduced amino acid sequence. A likely ribosomal binding site was identified 9bp upstream of the translational start site. The 3' end flank region encodes a peptide that shares high homology with dihydropteroate synthases. Nocardia GCH has 73 and 60% identity to the proteins encoded by the putative gch of Mycobacterium tuberculosis and Streptomyces coelicolor, respectively. Nocardia GCH was highly expressed in Escherichia coli cells carrying a pHAT10 based expression vector, and moderately expressed in Mycobacterium smegmatis cells carrying a pSMT3 based expression vector. Enterokinase digestion of recombinant Nocardia GCH, and in-gel digestion of Nocardia GCH and recombinant GCH followed by MALDI-TOF-MS analysis, confirmed that the actual subunit size of the enzyme was 24.5kDa. Thus, we conclude that the active form of native Nocardia GCH is a decamer. Our earlier incorrect conclusion was that the native enzyme was an octamer derived from the anomalous SDS-PAGE migration of the subunit. PMID- 15135391 TI - Vaccinia virus-mediated high level expression and single step purification of recombinant Jak2 protein. AB - Jak2 functions as a non-receptor tyrosine kinase and has been linked to pathologies such as cancer and cardiovascular disease. Because of this, many studies have tried to better understand its function. Unfortunately, very little information is known about its catalytic or biochemical properties as purification of significant amounts of functional Jak2 protein has been exceedingly difficult. Here, Jak2 was expressed in BSC-40 cells using a vaccinia virus-mediated expression system. Significant amounts ( approximately 10microg) of Jak2 protein were expressed from a single 100-mm cell culture dish. The protein was first harvested using three different methods of extraction to determine the relative efficiency of each lysis method with respect to Jak2 protein yield and catalytic activity. We found that lysis methods utilizing detergents increased the efficiency of protein extraction about 3-fold when compared to a method lacking detergent. However, with respect to catalytic activity, Jak2 isolated from cells using detergent-containing lysis buffers had significantly less catalytic activity than when compared to the method that was detergent free. Expression was then scaled up and Jak2 protein was purified via a one step immunoaffinity purification scheme using both the detergent-free and a modified detergent-containing method of extraction that maintained catalytic activity. In vitro kinase assays demonstrated that the purified product was highly catalytic as measured by its ability to tyrosine phosphorylate Stat1. Collectively, the results show that (1) Jak2 can be expressed at very high levels in mammalian cells, (2) it can be purified to homogeneity via a single step purification scheme, and (3) the purified product is biologically active. PMID- 15135392 TI - Purification of functional full-length liver X receptor beta produced in Escherichia coli. AB - Liver X receptor beta (LXRbeta) is a ligand dependent transcription factor that is a member of the nuclear receptor superfamily. LXRbeta and its isoform LXRalpha have recently been recognized as important regulators of lipid homeostasis in vertebrates. N-terminally hexahistidine-tagged rat LXRbeta was expressed in Escherichia coli as a full-length protein and purified in two chromatographic steps, immobilized metal affinity chromatography and gel filtration. From 10g of bacterial cells, 2.5mg of protein was recovered. The purified LXRbeta is functional with respect to ligand-, DNA-, and coactivator-binding. The synthetic ligand T0901317 bound to LXRbeta with high affinity yielding a K(d) of 2.7nM. Specific interaction with DR4 response elements, in the presence of RXR, was demonstrated with electrophoretic mobility shift assay. Furthermore, surface plasmon resonance analysis of LXRbeta binding to coactivator peptides revealed a ligand dependent interaction with the C-terminal nuclear receptor binding site of the coactivator RAP250. The purified LXRbeta constitutes an important tool for further functional and structural studies. PMID- 15135393 TI - A novel phospholipase A2/esterase from hyperthermophilic archaeon Aeropyrum pernix K1. AB - An open reading frame of the hyperthermophilic archaeon Aeropyrum pernix K1 APE2325, which composed of 474 bases, was cloned and expressed in Escherichia coli BL21 (DE3) Codon Plus-RIL. The recombinant protein was purified by Ni chelation affinity chromatography. It showed a single band with a molecular mass of 18kDa in SDS-PAGE. The purified enzyme exhibited both phospholipase A(2) and esterase activities with the optimal catalytic temperature at 90 degrees C. The enzyme activity was Ca(2+)-independent. Kinetic analysis revealed its Km, k cat, and Vm for the p-nitrophenyl propionate substrate were 103microM, 39s(-1), and 249micromol/min/mg, respectively. The recombinant protein was thermostable and its half-life at 100 degrees C was about 1h. PMID- 15135394 TI - High-yield expression and purification of p18 form of Bax as an MBP-fusion protein. AB - Bax is a pro-apoptotic member of the Bcl-2 family of proteins, which is present in the cytosol of various types of cells in full-length form (p21 Bax). During apoptosis, the N-terminal truncated version of Bax (p18 Bax) is often formed via cleavage of the p21 Bax by the calcium-dependent enzyme, calpain. p18 Bax is a membrane protein found primarily in the mitochondrial fraction of apoptotic cells. Although noticeable amounts of p18 Bax appear relatively late in apoptosis, it may still play a role in the apoptotic cascade. The role of p18 Bax in the apoptotic cascade, particularly, in the release of cytochrome c from mitochondria has not been studied. The goal of this study was to produce reasonable amounts of p18 Bax and study its effect on isolated mitochondria. The expression and purification of membrane proteins such as p18 Bax represents a substantial challenge due to insolubility. We report here that the apoptotic form of Bax, p18 Bax, is highly soluble in the absence of detergents upon fusion with maltose-binding protein (MBP). We describe a scheme for expression and simple metal-affinity based purification of MBP-p18Bax. The MBP-p18Bax triggers the release of cytochrome c from isolated mitochondria in a concentration-dependent, Bcl-2-sensitive manner. The MBP tag of the MBP-p18Bax can be cleaved off with 3C protease to produce pure p18 Bax, although the solubility of p18 Bax becomes very limited. The highly soluble, MBP-fused form of p18 Bax provides a convenient tool to study this apoptotic form of Bax protein. PMID- 15135396 TI - Cloning, expression, purification, and characterization of the human Class Ia phosphoinositide 3-kinase isoforms. AB - The Class I phosphoinositide 3-kinases (PI3Ks) are lipid kinases that phosphorylate the 3-hydroxyl group of the inositol ring of phosphatidylinositides. Although closely related, experimental evidence suggests that the four Class I PI3Ks may be functionally distinct. To further study their unique biochemical properties, the three human Class Ia PI3K (alpha, beta, and delta) p110 catalytic domains were cloned and co-expressed with the p85alpha regulatory domain in Sf9 cells. None of the p110 subunits were successfully expressed in the absence of p85alpha. Successful expression and purification of each p85alpha/p110 protein required using an excess of the p110 vector over the p85 vector during co-infection of Sf9 cells. Proteins were purified as the p85alpha/p110 complex by nickel affinity chromatography through an N-terminal His tag on the p110 subunit using an imidazole gradient. The purification yields were high using the optimized ratio of p85/p110 vector and small culture volumes, with 24mg/L cell culture media for p85alpha/p110alpha, 17.5mg/L for p85alpha/p110delta, and 3.5mg/L for p85alpha/p110beta. The identity of each purified isoform was confirmed by mass spectral analysis and immunoblotting. The activities of the three p85alpha/p110 proteins and the Class Ib p110gamma catalytic domain were investigated using phosphatidylinositol 4,5-bisphosphate (PIP2) as the substrate in a PIP2/phosphatidylserine (PS) liposome. All four enzymes exhibited reaction velocities that were dependent on the surface concentration of PIP2. The surface concentrations that gave maximal activity for each human isoform with 0.5mM PIP2 were 2.5mol% PIP2 for p110gamma, 7.5mol% for p85alpha/p110beta, and 10mol% PIP2 for p85alpha/p110alpha and p85alpha/p110delta. The specific activity of p85alpha/p110alpha was three to five times higher than that of the other human isoforms. These kinetic differences may contribute to the unique roles of these isoforms in cells. PMID- 15135395 TI - Cloning, expression, and purification of HLA-A2-BSP and beta-2m in Escherichia coli. AB - Tetramer analysis is a novel technique in immunological research that has dramatically changed our knowledge of the immune response to pathogens, tumors and autoimmune disease. Through the formation of major histocompatibility complex (MHC)-peptide tetrameric complexes, it can provide accurate counts of antigen specific T-cells and it allows their phenotypical and functional analysis. The tetramer is composed of the human leukocyte antigen (HLA) heavy chain, beta-2 microglobulin (beta-2m), the nominal peptide, and streptavidin. The HLA heavy chain and the beta-2m are expressed in Escherichia coli. But up to now, all laboratories have been expressing these two proteins by using isopropyl beta-d thiogalactopyranoside IPTG. IPTG is very expensive, and it is tedious and laborious to induce expression protein. So it is difficult to scale up to express the objective protein. To address this problem, extracellular fractions of HLA A0201 and beta-2m (absent signal peptide) genes were cloned from peripheral blood mononuclear cells (PBMCs) by RT-PCR. DNA coding for a Gly-Ser linker and a BSP (15-amino acid substrate peptide for BirA-dependent biotinylation) was added to the COOH-terminus of the extracellular fraction of HLA-A0201 by PCR, using an HLA A0201 as the template. Then the HLA-A0201-BSP and beta-2m genes were cloned into pBV220 vector and expressed, respectively. The expressed proteins were purified and detected by ELISA and Western blot analyses. High-efficient expressions of HLA-A0201-BSP and beta-2m proteins lay a good foundation for further expression and purification in prokaryotic system and constructing MHC class I-peptide tetramer complexes to study the function of CTLs. PMID- 15135397 TI - Expression, refolding, and purification of recombinant human phosphodiesterase 3B: definition of the N-terminus of the catalytic core. AB - We have developed an expression, refolding, and purification protocol for the catalytic domain of human Phosphodiesterase 3B (PDE3B). High level expression in Escherichia coli has been achieved with yields of up to 20mg/L. The catalytic domain of the enzyme was purified by affinity chromatography utilizing a novel affinity ligand. PDE3B, purified by affinity chromatography, with no single impurity #10878;1% as determined by SDS-PAGE, has a specific activity of 2210+/ 442nmol/min/mg and a KM for cAMP of 44+/-4.5nM. Reducing the size of the expressed catalytic domain from residues 387-1112 to residues 654-1086 greatly reduced the aggregation phenomena observed with the affinity purified PDE3B. The definition of the N-terminus of the catalytic core was examined through the generation of several truncation mutants spanning amino acid residues 636-674. Constructs starting at E665 and M674 were fully active and devoid of activity, respectively. A construct starting at D668 had a Vmax reduced by approximately 10 fold relative to the longer constructs, yet the KM was not affected. This indicates the minimal N-terminus of the catalytic core lies between E665 and Y667. Refolding and affinity purification of the 654-1073 catalytic core of PDE3B has been employed to produce large quantities of highly pure enzyme for structural studies. PMID- 15135398 TI - Yersinia enterocolitica type III secretion chaperone SycH. Recombinant expression, purification, characterisation, and crystallisation. AB - All pathogenic Yersinia species (Y. enterocolitica, Y. pestis, and Y. pseudotuberculosis) share a type three secretion system (TTSS) that allows translocation of effector proteins into host cells. Yersinia enterocolitica SycH is a chaperone assisting the transport of the effector YopH and two regulatory components of the TTSS, YscM1 and YscM2. We have recombinantly expressed SycH in Escherichia coli. Purification of tag-free SycH to near homogeneity was achieved by combining ammonium sulfate precipitation, anion exchange chromatography, and gel filtration. Functionality of purified SycH was proven by demonstrating binding to YopH. SycH crystals were grown that diffracted to 2.94A resolution. Preliminary crystallographic data and biochemical findings suggest that SycH forms homotetramers. SycH may therefore represent a novel class of TTSS chaperones. In addition, we found that YopH was enzymatically active in the presence of SycH. This implies that the function of the secretion chaperone SycH is not to keep YopH in a globally unfolded state prior to secretion. PMID- 15135399 TI - Subunit protein-affinity isolation of Drosophila DNA polymerase catalytic subunit. AB - gfLittle is known at present about the biochemical properties of very large-sized Drosophila DNA polymerases. In a previous study, we tried to purify Drosophila pol. catalytic subunit from embryos through seven column chromatographies and study its biochemical properties. However, we failed to characterize it precisely because an insufficient amount of the enzyme was generated. In this report, we describe direct purification from Drosophila embryos to near homogeneity using Drosophila DNA polymerase second subunit (Drosophila pol. 2) protein-conjugated affinity column chromatography and characterization of the enzyme in detail. To our knowledge this is the first demonstration of native DNA polymerase purification with activity using a subunit protein-affinity column. We observed new characteristics of Drosophila pol. catalytic subunit as follows: Drosophila pol. catalytic subunit synthesized DNA processively in the presence of both Mn(2+) and Mg(2+) ions, but Mn(2+) inhibited the 3'-5' proofreading activity, thereby decreasing the fidelity of DNA replication by 50%. PMID- 15135400 TI - Expression, purification, and antifreeze activity of carrot antifreeze protein and its mutants. AB - Antifreeze proteins (AFPs) enable organisms to survive under freezing or sub freezing conditions. AFPs have a great potential in the low temperature storage of cells, tissues, organs, and foods. This process will require a large number of recombinant AFPs. In the present study, the recombinant carrot AFP was highly expressed in Escherichia coli strain BL21 (DE3). The activity of the purified and refolded recombinant proteins was analyzed by measurement of thermal hysteresis (TH) activity and detection of in vitro antifreeze activity by measuring enhanced cold resistance of bacteria. Two carrot AFP mutants generated by site-directed mutagenesis were also expressed and purified under these conditions for use in parallel experiments. Recombinant DcAFP displayed a TH activity equivalent to that of native DcAFP, while mutants DcAFP-N130Q and rDcAFP-N130V showed 32 and 43% decreases in TH activity, respectively. Both the recombinant DcAFP and its mutants were able to enhance the cold resistance of bacteria, to degrees consistent with their respective TH activities. PMID- 15135401 TI - Isolation, cloning, and overexpression of a chitinase gene fragment from the hyperthermophilic archaeon Thermococcus chitonophagus: semi-denaturing purification of the recombinant peptide and investigation of its relation with other chitinases. AB - A 189-bp sequence was isolated from the hyperthermophilic archaeon Thermococcus chitonophagus and was found to present strong homology with a large number of chitinase genes from a variety of organisms and particularly with the chitinaseA gene from Pyrococcus kodakaraensis (Pk-chiA). This fragment was subcloned to an expression vector and overexpressed in Escherichia coli. The E. coli BLR21(DE3)pLysS transformant, harbouring the gene on the pET-31b plasmid vector, was found to overproduce the target protein at high levels. The 63 aminoacid-long peptide was efficiently purified to homogeneity, with a one-step, semi-denaturing affinity chromatography, on a metal chelation resin and was used for the production of a specific, polyclonal antibody from rabbits. The produced antibody was demonstrated to display strong and specific affinity for the chitinase A from Serratia marcescens (Sm-chiA), as well as the membrane-bound chitinase70 from Thermococcus chitonophagus (Tc-Chi70). The strong sequence homology, in combination with the demonstrated specific immunochemical affinity, indicates that the isolated peptide is part of a chitinolytic enzyme of T. chitonophagus. In particular, it could belong to the membrane-bound chi70, or to a distinct chitinase, coded by a different gene, or even by the same gene, following post transcriptional or post-translational modifications. PMID- 15135402 TI - Purification and characterization of inulinase from Aspergillus niger AF10 expressed in Pichia pastoris. AB - The inuA1 gene encoding an exoinulinase from Aspergillus niger AF10 was expressed in Pichia pastoris, and the recombinant enzyme activity was 316U/ml in a 5L fermentor, with the inulinase protein accounting for 35% of the total protein of fermentation broth. The hydrolysis rate of inulin can reach 92%, with a 25U/g inulin enzyme addition, and 90% of fructose content after 6h. Glucose can significantly inhibit the enzymatic hydrolysis of inulin. This is the first report of glucose inhibition of inulinase-catalyzed hydrolysis. PMID- 15135403 TI - Cloning, expression, and purification of 5,10-methenyltetrahydrofolate synthetase from Mus musculus. AB - Folate metabolism is necessary for the biosyntheses of purine nucleotides and thymidylate and for the synthesis of S-adenosylmethionine, a cofactor required for cellular methylation reactions and a precursor of spermidine and spermine syntheses. Disruption of folate metabolism is associated with several pathologies and developmental anomalies including cancer and neural tube defects. The enzyme 5,10-methenyltetrahydrofolate synthetase (MTHFS, EC 6.3.3.2) catalyzes the ATP dependent conversion of 5-formyltetrahydrofolate to 5,10 methenyltetrahydrofolate, and has been shown to affect intracellular folate concentrations by accelerating folate degradation. Mammalian MTHFS proteins described to date are not stable and no recombinant mammalian MTHFS protein has been successfully expressed in Escherichia coli. The three-dimensional structure of MTHFS has not been solved. The cDNA coding for Mus musculus MTHFS was isolated and expressed in E. coli with a hexa-histidine tag. Milligram quantities of recombinant mouse MTHFS were purified using metal affinity chromatography and the protein was stabilized with Tween 20. Mouse MTHFS has a molecular mass of 23kDa and is 84% identical in amino acid sequence to the human enzyme. Activity assays confirmed the functionality of the recombinant protein, with Km =5 microM for (6S)-5-formyltetrahydrofolate and Km=769 microM for Mg-ATP. This is the first example of a mammalian form of MTHFS expressed in E. coli that yielded sufficient quantities of stable purified protein to allow for detailed characterization of its three-dimensional structure and kinetic properties. PMID- 15135404 TI - Recombinant human serum amyloid P component from Pichia pastoris: production and characterization. AB - Human serum amyloid P component (SAP) was expressed in the methylotrophic yeast Pichia pastoris. SAP cDNA was placed under control of regulatory sequences derived from the alcohol oxidase gene (AOX1), and its protein product was secreted using the Saccharomyces cerevisiae alpha-mating factor signal sequence. Recombinant SAP (r-SAP) was produced in a bioreactor with computer controlled fed batch mode and purified by use of a C-terminal histidine tag. The yield of purified r-SAP was 3-4mg from 1L supernatant and 5-6mg from 1L cell paste, indicating that the majority of the produced SAP was not secreted. Treatment of the cell paste with EDTA increased the yield further by about 30%. The N-terminal of r-SAP purified from the supernatant showed non-complete cleavage of the alpha mating factor signal sequence. Purified r-SAP, analyzed under native conditions, was shown to be a decamer, like purified human SAP (h-SAP), with monomers of 27kDa. Each monomer had one N-glycosylation site, positioned at the same site as for h-SAP. r-SAP bound to antibodies produced against h-SAP. Furthermore, r-SAP bound to ds DNA and influenza A virus subunits in a Ca(2+)-dependent manner and inhibited influenza A virus hemagglutination. These results indicate that r-SAP produced in P. pastoris has the same biological activity as purified h-SAP. PMID- 15135405 TI - In vivo increase of solubility of overexpressed Hha protein by tandem expression with interacting protein H-NS. AB - Gene cloning in appropriate vectors followed by protein overexpression in Escherichia coli is the common means for protein purification. This approach has many advantages but also some drawbacks; one of these is that many proteins fail to achieve a soluble conformation when overexpressed in E. coli. Hha protein belongs to a family of nucleoid-associated proteins functionally related to the H NS family of proteins. Hha-like proteins and H-NS-like proteins are able to semidirectly bind to each other. We show in this work that overexpressed Hha or HisHha protein (a functional derivative of Hha containing a 6x His tag at the amino end) from a T7-polymerase promoter in BL21 DE3 E. coli strains results in the vast majority of the protein accumulated in insoluble aggregates (inclusion bodies). We also show that tandem overexpression of HisHha and H-NS increases the solubility of HisHha and prevents the formation of inclusion bodies. Single amino acid substitutions in the HisHha protein, which impair interaction with H-NS, render insoluble protein even when tandem-expressed with H-NS, tandem expression of an insoluble protein and an interacting partner is an experimental strategy which could be useful to increase the solubility of other overexpressed proteins in E. coli. PMID- 15135406 TI - Expression, purification, and DNA-binding activity of the Herbaspirillum seropedicae RecX protein. AB - The Herbaspirillum seropedicae RecX protein participates in the SOS response: a process in which the RecA protein plays a central role. The RecX protein of the H. seropedicae, fused to a His-tag sequence (RecX His-tagged), was over-expressed in Escherichia coli and purified by metal-affinity chromatography to yield a highly purified and active protein. DNA band-shift assays showed that the RecX His-tagged protein bound to both circular and linear double-stranded DNA and also to circular single-stranded DNA. The apparent affinity of RecX for DNA decreased in the presence of Mg(2+) ions. The ability of RecX to bind DNA may be relevant to its function in the SOS response. PMID- 15135407 TI - Expression and purification of untagged full-length HCV NS5B RNA-dependent RNA polymerase. AB - The NS5B encoded by the hepatitis C virus genome is a RNA-dependent RNA polymerase essential to viral replication. The entire NS5B protein contains a catalytic domain followed by a regulatory motif and a membrane-anchor domain at its C-terminus. Reported here is the molecular cloning and expression of the full length NS5B polymerase (NS5B-FL) in bacterial cells as a non-fusion protein. The non-tagged NS5B-FL was purified to homogeneity using sequential chromatographic columns and its identity was confirmed using anti-NS5B peptide antibodies and amino acid sequencing. Purified NS5B-FL demonstrated RNA-dependent RNA polymerase activity and was able to replicate a HCV RNA genome fragment through both copy back and de novo mechanisms. Its biochemical properties were further characterized in comparison with a truncated form of NS5B polymerase with a deletion of 51 residues from its C-terminus. PMID- 15135409 TI - Expression and purification of His-tagged rat mitochondrial 3-ketoacyl-CoA thiolase wild-type and His352 mutant proteins. AB - Mitochondrial 3-ketoacyl-CoA thiolase is a key enzyme for the beta-oxidation of fatty acids, and the deficiency of this enzyme in patients has been previously reported. We cloned a cDNA of rat mitochondrial 3-ketoacyl-CoA thiolase into a bacterial expression vector pLM1 with six continuous histidine codons attached to the 5' end of the gene. The cloned cDNA was overexpressed in Escherichia coli and the soluble protein was purified with a nickel Hi-Trap chelating metal affinity column in 92% yield to apparent homogeneity. The specific activity of the purified His-tagged rat mitochondrial 3-ketoacyl-CoA thiolase was 25U/mg. It has been proposed that His352 is a catalytic residue responsible for activation of coenzyme A by deprotonation of a sulfhydryl group. We constructed four mutant expression plasmids of the enzyme using site-directed mutagenesis. Mutant proteins were overexpressed in E. coli and purified with a nickel metal affinity column. Kinetic studies of wild-type and mutant proteins were carried out, and the result confirmed that His352 is a catalytic residue of rat mitochondrial 3 ketoacyl-CoA thiolase. Our overexpression in E. coli and one-step purification of the highly active rat mitochondrial 3-ketoacyl-CoA thiolase greatly facilitated our further investigation of this enzyme, and our result from site-directed mutagenesis increased our understanding of the mechanism for the reaction catalyzed by 3-ketoacyl-CoA thiolase. PMID- 15135408 TI - Synthesis and assembly of a cholera toxin B subunit SHIV 89.6p Tat fusion protein in transgenic potato. AB - A cDNA encoding the simian-human immunodeficiency virus (SHIV 89.6p) Tat regulatory element protein was fused to the c-terminus of the cholera toxin B subunit gene (ctxB-tat) and introduced into Solanum tuberosum cells by Agrobacterium tumefaciens-mediated transformation methods. The fusion gene was detected in the genomic DNA of transformed potato leaf cells by PCR DNA amplification. Synthesis and assembly of the CTB-Tat fusion protein into oligomeric structures of pentamer size was detected in transformed tuber extracts by immunoblot analysis. The binding of CTB-Tat fusion protein pentamers to intestinal epithelial cell membrane glycolipid receptors was quantified by G(M1) ganglioside enzyme-linked immunosorbent assay (G(M1)-ELISA). Based on the ELISA results, CTB-Tat fusion protein made up about 0.005-0.007% of total soluble tuber protein or approximately 4.6mg per 100g potato tuber tissue. The synthesis and assembly of CTB-Tat monomers into biologically active oligomers in transformed potato tuber tissues demonstrates the feasibility of using viral pathogen antigens synthesized in edible plants for mucosal immunization against HIV-1 infection. PMID- 15135410 TI - Bacterially expressed recombinant p68 RNA helicase is phosphorylated on serine, threonine, and tyrosine residues. AB - We previously reported the expression and purification of recombinant p68 RNA helicase in a bacterial expression system. The recombinant p68 is an RNA dependent ATPase and ATP-dependent RNA helicase. In the process of characterizing the ATPase and RNA unwinding activities of the recombinant p68, we observed that the bacterially expressed p68 RNA helicase is phosphorylated on tyrosine, serine, and threonine residues. Our data demonstrated that phosphorylations on the recombinant p68 RNA helicase affect the enzymatic activities of the protein. This is the first observation that recombinant protein expressed in bacteria Escherichia coli is phosphorylated at multiple residues by bacterial endogenous protein kinases. Our observations suggest an important mechanism in controlling the function of p68 RNA helicase by signal transduction pathways. PMID- 15135411 TI - Expression, purification, and in vitro characterization of recombinant salmon insulin-like growth factor-II. AB - The insulin-like growth factors, IGF-I and IGF-II, are single chain polypeptides, which are structurally related to proinsulin and promote proliferation and differentiation of cells in many vertebrate species. Previous attempts to produce recombinant salmon IGF-II (rsIGF-II) were compromised by low expression levels and co-purification of incorrectly cleaved protein with the authentic recombinant product. In this study, a gene containing the coding region for Atlantic salmon (Salmo salar) IGF-II was cloned into a modified pET32a expression vector and transformed into Escherichia coli BL21 trxB (DE3) cells. Upon growth and induction (with IPTG) of the transformant, recombinant salmon IGF-II (rsIGF-II) was expressed as an insoluble, 28kDa thioredoxin.sIGF-II fusion protein linked by a protease cleavage motif (trx.FAHY.sIGF-II) in inclusion bodies. The inclusion bodies were subsequently solubilized and the fusion protein was purified by Ni affinity chromatography. Recombinant IGF-II (7.8kDa) was then released from the fusion partner using H64A subtilisin BPN' protease and purified by reversed-phase HPLC. Homogeneity of the final recombinant product was confirmed by N-terminal amino acid sequencing, ion-spray mass spectrometry, SDS-polyacrylamide gel electrophoresis, and analytical reversed-phase HPLC. The biological activity of rsIGF-II was demonstrated in cultured rat L6 myoblasts and was found to be approximately 9- and 5-fold less potent than recombinant human IGF-I and recombinant salmon IGF-I, respectively, a result similar to that demonstrated previously with other recombinant fish IGF-II's in non-homologous cell lines. PMID- 15135413 TI - Overexpression of a synthetic gene encoding human alpha interferon in Escherichia coli. AB - Interferons (IFNs) represent an important defense mechanism in vertebrates. In this work, we describe gene synthesis and assembly using the polymerase chain reaction as a method for single-step synthesis of DNA sequences. The oligonucleotides designed were based on Escherichia coli codon usage and two genes of IFN were synthesized: one containing a DNA sequence already known and the other, a mutated form in which two cysteine amino acid residues were replaced by serines in an attempt to improve the stability of the protein. DNA sequences were cloned into pAE, an E. coli vector that allows heterologous protein expression with or without a histidine tag. Recombinant human interferons (rhIFNs) were identified by Western blotting and ELISA using anti-human interferon polyclonal antibodies. Purification of the recombinant His-tagged proteins was achieved in a single step by Ni(2+)-charged column chromatography while proteins without His-tag were purified by extensively washing the inclusion bodies, the final yields being approximately 210 and 75mg/L, respectively. The rhIFNs expressed within this system were biologically active ( approximately 1,1x10(8)IU/mg) based on antiviral assay. The combined methodologies described here proved to be cost-effective and could be extended to other genes/proteins of interest. PMID- 15135412 TI - Cloning, expression, and structural analysis of recombinant BJcuL, a c-type lectin from the Bothrops jararacussu snake venom. AB - The lactose-binding lectin from Bothrops jararacussu venom (BJcuL) is a homodimer belonging to group VII of the c-type animal lectins. BJcuL has also been shown to serve as an interesting tool for combating tumor progression by inhibiting cancer and endothelial cell growth. However, detailed structural studies of BJcuL and its biological mechanisms of cytotoxicity are yet to be reported, perhaps because of the non-availability of recombinant proteins in necessary quantities. Intending to increase the present information about structural and consequently the understating of biological studies, the cDNA coding for BJcuL from a venom gland has been cloned and sequenced. The mature protein-coding region was amplified by PCR with specific oligonucleotides, and subcloned into the pET-15b vector to express the recombinant BJcuL in Escherichia coli BL21 (DE3). The deduced amino acid sequence exhibits a high degree of sequence identity with c type lectins (CTLs) and c-type lectin-like domains (CTLDs). An insoluble and inactive 18.5-kDa protein was overexpressed after 1.0mM IPTG induction. The recombinant BJcuL was recovered and denatured in a buffer with 6M urea and purified on a nickel-affinity column. Protein refolding was carried out on this column, during procedure purification, followed by dialysis against CTBS and then by gel filtration for separation of the active dimmer. The refolding process of rBJcuL and the analysis of its structure were confirmed by biological assay, circular dichroism, and MALDI-TOF. PMID- 15135414 TI - Purification and refolding of human alpha5-subunit (PSMA5) of the 20S proteasome, expressed as inclusion bodies in Escherichia coli. AB - The 20S proteasome is the central enzyme of nonlysosomal protein degradation in both the cytosol and nucleus. It is composed of 28 protein subunits which are arranged into four staggered heptameric rings. The outer rings consist of alpha subunits which are responsible for binding of proteasome activators, inhibitors, and regulators. To better characterize human alpha5-subunit (PSMA5) of the 20S proteasome, we have established a high-efficiency Escherichia coli expression system. The DNA-coding sequence for the human PSMA5, which was subcloned into the vector pET-22b (+), has been expressed as inclusion bodies in E. coli BL21 (DE3). To produce the native PSMA5, straightforward protocols have been developed for refolding the human PSMA5 in the presence of surfactants using dilution refolding and size-exclusion chromatography matrix refolding methods. After refolding, recovery yields of about 20% were obtained, respectively, with purity above 95%. The human PSMA5 was detected by dynamic light scattering in refolding process, and the molecular weight of the final refolded product was measured using gel filtration chromatography, which indicates that the human PSMA5 exists mainly as tetramer. PMID- 15135415 TI - Rapid purification of truncated tau proteins: model approach to purification of functionally active fragments of disordered proteins, implication for neurodegenerative diseases. AB - Truncated tau is of great interest because of its important role in neurofibrillary pathogenesis in Alzheimer's disease (AD). A major obstacle for characterization of detailed biochemical and biological properties of truncated tau species and their fragments has been the lack of reliable and quick purification methods. Uneven distribution of acidic and basic residues in tau determines that the N- and C-terminal tau fragments require entirely different purification conditions. Conventional methods take several days; they do not allow purification of the acidic N-terminal tau fragments and do not prevent aggregation during purification that makes purified truncated tau unusable in functional studies. To prevent these inherent problems, we have designed a two step, highly efficient purification procedure yielding a fully functional, non aggregated homogeneous population of truncated tau molecules. Various forms of tau produced in bacteria without the need for a heat pre-treatment step were subjected to anion- and cation-exchange chromatography. Conditions were developed that allowed effective separation and purification of acidic and/or basic tau species. Following the gel filtration step, up to 10mg of tau proteins with 96% purity was obtained within one working day. Purified truncated tau exhibited an unmodified immunoreactivity and allowed its functional activity analysis. Since many neurodegenerative diseases have implicated similar disordered proteins in their pathogenesis, our procedure will allow their detailed analysis and characterization. PMID- 15135416 TI - Isolation of aminoglycoside nucleotidyltransferase (2'')-Ia from inclusion bodies as active, monomeric enzyme. AB - Aminoglycoside nucleotidyltransferase( 2'')-Ia (ANT( 2'') confers resistance to pathogenic bacteria against several aminoglycoside antibiotics including gentamicin, kanamycin, and tobramycin. The gene for this aminoglycoside-modifying enzyme has been cloned from a clinical isolate of Pseudomonas aeruginosa. This gene was inserted into an overexpression vector, the vector was then transformed into Escherichia coli BL21(DE3), and the protein has been isolated in the form of inclusion bodies. Optimal refolding conditions have been determined to be direct dilution of solubilized inclusion bodies into 0.1M Tris-HCl, pH 8.5, 0.2M KCl, 0.4M l-arginine, and 5mM reduced glutathione at 4 degrees C. The refolded enzyme is monomeric in solution and has similar kinetic properties and substrate selectivity to the enzyme isolated in soluble form. PMID- 15135417 TI - Pasteurization of antithrombin without generation of the prelatent form of antithrombin. AB - Human antithrombin (AT) is the major inhibitor of blood coagulation and has also been shown to exert anti-inflammatory and anti-angiogenic effects. Pasteurization of pharmaceutical AT products is usually performed at 60 degrees C for 10h in the presence of sodium citrate as stabilizer, sometimes in combination with sucrose. These stabilizers significantly decrease the aggregation and denaturation of AT, but during the pasteurization, a small amount of latent AT (LAT), a partially denatured form, is usually generated, as is an equal amount of another latent form of AT, the so-called prelatent AT (PLAT). The LAT formed during pasteurization has a rather low affinity to heparin and is easily removed by using a second heparin affinity chromatography step in the production process. This is in contrast to the PLAT, which has a slightly lower affinity to heparin than does native AT, which makes it hard to remove. Hence, four commercial products of pasteurized AT were previously shown to contain about 4% of PLAT. In the present work, an alternative pasteurization method is presented, where 2M ammonium sulfate and 50% sucrose are used as stabilizers. During this pasteurization, no, or trace amounts ( < 0.5%), of PLAT may be generated with no formation of aggregates. Moreover, the pasteurized AT has the same specific thrombin-inhibiting activity when compared to incubation in the presence of citrate and sucrose. Heparin affinity high-performance liquid chromatography was used for the determination of PLAT, LAT, and AT. PMID- 15135418 TI - UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase, functionally expressed in and purified from Escherichia coli, yeast, and insect cells. AB - UDP-GlcNAc 2-epimerase/ManNAc kinase is the key enzyme of sialic acid biosynthesis in mammals. Its functional expression is a prerequisite for early embryogenesis and for the synthesis of several cell recognition motifs in adult organism. This bifunctional enzyme is involved in the development of different diseases like sialuria or hereditary inclusion body myopathy. For a detailed understanding of the enzyme, large amounts of the pure active protein are needed. Different heterologous cell systems were therefore analyzed for the enzyme, which was found to be functionally expressed in Escherichia coli, the yeast strains Saccharomyces cerevisiae and Pichia pastoris, and insect cells. In all these cell types, the expressed enzyme displayed both epimerase and kinase activities. In E. coli, up to 2mg protein/l cell culture was expressed, in yeast cells only 0.4mg/L, while up to 100mg/L, were detected in insect cells. In all three cell systems, insoluble protein aggregates were also observed. Purification from E. coli resulted in 100microg/L pure and structurally intact protein. For insect cells, purification methods were established which resulted in up to 50mg/L pure, soluble, and active protein. In summary, expression and purification of the UDP GlcNAc 2-epimerase/ManNAc kinase in Sf-900 cells can yield the milligram amounts of protein required for structural characterization of the enzyme. However, the easier expression in E. coli and yeast provides sufficient quantities for enzymatic and kinetic characterization. PMID- 15135420 TI - [Faecal inflammatory markers in nutrition and digestive diseases in children]. PMID- 15135419 TI - Optimised expression in Escherichia coli and purification of the functional form of the Bacillus thuringiensis Cry4Aa delta-endotoxin. AB - Achieving high-level expression of the Bacillus thuringiensis Cry4Aa mosquito larvicidal protein was demonstrated. The 130-kDa Cry4Aa protoxin was overexpressed as an inclusion body in Escherichia coli under the control of the tac promoter together with the cry4Ba promoter. The solubility of the toxin inclusions in carbonate buffer, pH 10.0, was markedly enhanced at a cultivation temperature of 30 degrees C. Elimination of the tryptic cleavage site at Arg-235 in the loop between helices 5 and 6 still retained the high-level toxicity of E. coli cells expressing the Cry4Aa mutant against Aedes aegypti larvae. Trypsin digestion of the R235Q mutant protoxin produced a protease-resistant fragment of ca. 65kDa. A homogeneous product of the 65-kDa trypsin-treated R203Q protein was obtained after size-exclusion chromatography that would pave the way for the further crystallisation and X-ray crystallographic studies. PMID- 15135421 TI - [Acute uncomplicated malaria treatment in children in France in 2002]. AB - CONTEXT AND OBJECTIVES: Imported P. falciparum malaria cases are rising in France reaching 1300 estimated children in 2000. Three years after the publication of therapeutic guidelines, the Groupe de Pediatrie Tropicale conducted an observational survey in order to describe the practice of acute uncomplicated malaria treatments in children, to identify their limits and to make proposals to improve them. METHODS: A self administrated questionnaire has been proposed to 29 pediatric wards declaring over 10 malaria cases in 2000. Questions were focused on treatment practices and monitoring of children diagnosed with acute uncomplicated malaria in 2002. RESULTS: Twenty-six services, who treated more than 700 children, responded. Twenty-two on 26 services hospitalized systematically malaria cases. Mean duration of hospitalization was 2.2 days (S.D. +/- 0.9). First line treatment was halofantrine in 22 on 26 services and mefloquine in four services. A second halofantrine dose was given systematically at day 7 in three services. No clinical cardiac effects happened. Quinine was used in perfusion only in cases of gastric intolerance. Treatment failure has never been experienced in the 22 services using halofantrine and has been experienced at least once in two on four wards using mefloquine. Relapse occurred at least once in 19 on 22 wards treating with halofantrine. DISCUSSION: Halofantrine with hospitalization is still the leading treatment of acute uncomplicated malaria in children in France. In spite of the absence of clinical cardiac incident, a second cure of halofantrine was not often used, exposing to a high rate of relapse. Mefloquine is three time more used than in 1997, in spite of its digestives side effects that can explain failures of treatment. When done, the systematic clinical and parasitological control confirms failures after mefloquine and the high incidence of relapse after one cure of halofantrine. Only used in case of severity or digestive disorders, quinine is a little less prescribed in acute uncomplicated malaria in 2001 than in 1997. CONCLUSIONS: The limitations of antimalarial drugs used in France in case of acute malaria argue for an improvement of protocols (systematic second reduced dose of halofantrine after day 7, mefloquine associated with antiemetic drug) and a systematic clinical and parasitological monitoring. As alternative, efficient combinations of antimalarial as first line treatment are needed in France. PMID- 15135422 TI - [Very preterm infant (< 32 weeks) vs very low birth weight newborns (1500 grammes): comparison of two cohorts]. AB - Studies concerning very preterm newborns are either defined by birth weight (<1500 g) or gestational age (<32 weeks). The aim of our study was to underline limits of cohort definitions by birth weight. METHODS: Data come from the Nord Pas de Calais EPIPAGE cohort. Every birth occurring in 1997 before 32 weeks or with a birth weight less than 1500 g and transferred in a neonatal unit was included. Two cohorts were defined, one by gestational age (<32 weeks), the other by birth weight (<1500 g). Two subgroups could be defined from these to cohorts: group A (<32 weeks and > or =1500 g), from cohort (<32 weeks), group B (> or =32 weeks and <1500 g) from cohort (<1500 g). RESULTS: Five hundred nine newborns were included. Perinatal characteristics of both cohorts seemed comparable. The analysis by subgroups A and B revealed an excess of pulmonary and neurological morbidity in very preterm infant compared to very low birth weight newborn. This was linked to an excess of growth restricted newborns in this cohort with more advanced gestational ages. CONCLUSION: Cohorts of very preterm newborns should rather be defined by gestational age. If not possible, results in very low birth weight cohorts should also be given by gestational age and rate of growth restriction should be described. PMID- 15135423 TI - [Meconium aspiration syndrome requiring mechanical ventilation: incidence and respiratory management in France (2000-2001)]. AB - OBJECTIVES: To precise the number of term neonates with a meconium aspiration syndrome requiring mechanical ventilation in 2000 and 2001 in continental France and the related mortality. To study the different respiratory management. METHOD: A written questionnaire was sent to all intensive care units of continental France. RESULTS: Thirty-nine units were contacted and 31 answered (80%). On 30 answering centers, 265 cases are reported in 2000 and 249 in 2001. Extrapolation to the 39 centers led to a number of 347 cases in 2000 and 324 in 2001. Mortality rate was 6.4% for 2 years. Causes of death were: neurological 61%, respiratory 18% and others 21%. Twenty-six on 29 units used exogenous surfactant and/or high frequency oscillatory ventilation (HFOV) and/or inhaled nitric oxide (iNO). The sequence of use and the indication criteria of these therapies were quite variable with a majority of units using HFOV in the first place. CONCLUSIONS: Meconium aspiration syndrome is still a frequent neonatal condition despite a codified prevention strategy. Respiratory management in intensive care units remains very variable. PMID- 15135425 TI - [Isolated familial corpus callosum agenesis prognosis]. AB - Agenesia of corpus callosum belongs to a group of cerebral malformations whose prognosis is uncertain. In such cases, assessment of prognosis may benefit from eventual associated fetal, obstetrical or familial features. We report a patient with an isolated corpus callosum agenesia that led to the discovery of a similar malformation in her father. This observation demonstrates that some forms of isolated and familial corpus callosum agenesia could have a favorable outcome. However, the difficulty of the assessment of prognosis in isolated corpus callosum agenesia is emphasized and the question of parental RMI exploration in such a peculiar context is raised. PMID- 15135424 TI - [Parent's information and prenatal diagnosis of cerebral malformation with an uncertain prognosis]. AB - Fetal ultrasounds examinations allow identification of brain malformations; announce of diagnosis and information about prognosis may be difficult when malformation is rare and prognosis uncertain. OBJECTIVES: In this study we tried to analyze how fetal imaging for prenatal screening was organized and how couples were managed and supported. METHODS: We focused on the procedures used to inform couples: content, method of delivery and consequences. A referent physician in each large multidisciplinary center for prenatal diagnosis in Paris area was questioned by semi-directed interview. RESULTS: Our study showed that it is difficult to standardize the way in which information is supplied before and after fetal ultrasounds examination; uncertainty about prognosis led more often to abortion. CONCLUSION: Thus, couples should have clear and complete information provided by a multidisciplinary team including specialists particularly concerned by the malformation (neuropediatrician and/or neurosurgeon)--moreover when prognosis is uncertain, in order to support them, and to accompany their decision concerning pregnancy. PMID- 15135426 TI - [Cardiorespiratory arrest in full term newborn infants: six case reports]. AB - Cardiorespiratory arrest occurring within the first two hours of life of a perfectly normal newborn is a very seldom event hitherto unreported. Six infants born after an uneventful pregnancy by normal vaginal delivery, with a normal Apgar score and physical examination, were found with unexpected cardiorespiratory arrest requiring cardiac and respiratory resuscitation early after birth. All were lying in the prone position, their face covered up while facing mother's abdomen, breast or neck. All mothers were primipara. All newborns but one died. Biological and bacteriological samples were normal and early onset neonatal sepsis was ruled out. Autopsy, performed in five infants, was not contributive. We hypothesize that the sudden and unexpected cardiorespiratory arrest occurring in these normal newborns was secondary to acute upper airway obstruction. To prevent this life threatening post-natal asphyxic episode, it is essential to ensure that the face of a newborn lying down upon mother's breast and abdomen is properly and continuously cleared. PMID- 15135428 TI - [Childhood hypophosphatasia: a case report due to a novel mutation]. AB - Hypophosphatasia is characterized by defective bone mineralization associated with impaired activity of the tissue non-specific alkaline phosphatase (TNSALP) due to mutations in the TNSALP gene. We describe a child with a mutation that has not been described up to now. CASE REPORT: A 4-year-old child presented with clinical symptoms of rickets and premature loss of decideous teeth. Reduced serum alkaline phosphatase activity and radiographic features led to the diagnosis of hypophosphatasia, which was confirmed by genetic investigation. The molecular study showed two missense mutations, of which one is a novel mutation. CONCLUSION: Hypophosphatasia is suspected in a child with rickets and premature loss of decideous teeth. Such symptoms should prompt the search of a reduced serum alkaline phosphatase activity. The clinical and molecular diagnosis of the disease is important for the genetic counseling but also for a proper determination of prognosis, as it is related to the type of mutation. PMID- 15135427 TI - [Sudden death of neonates in the delivery room]. AB - Healthy newborn death in the delivery room is uncommon. Unlike for sudden infant death syndrome well described in infants between 2 and 6 months of age, few publications have studied this event. We report two cases of asymptomatic term newborns who died unexpectedly in the delivery room. Noteworthy, these newborns were sleeping in prone position on their mother. The Agence Nationale d'Accreditation et d'Evaluation en Sante (ANAES) published recommendations to promote breast-feeding including uninterrupted early contact between the infant and his mother. However, immediately after birth, the newborn may be particularly vulnerable. The application of this recommendation unwisely could be dangerous for newborns allowed to stay on their mother without any monitoring, or medical supervision. We would like to point out the importance of healthy newborn supervision within the first hours of life that can be done without interfering with the mother-child bonding. PMID- 15135429 TI - [Genetics of hypophosphatasia]. AB - Hypophosphatasia is a rare inherited disorder characterized by defective bone and teeth mineralization and deficiency of serum and bone alkaline phosphatase activity. The symptoms are highly variable in their clinical expression, which ranges from stillbirth without mineralized bone to early loss of teeth without bone symptoms. Currently, there is no treatment for the disease. Recent developments in molecular biology allow to better understand the genetics of the disease, especially its transmission that may be recessive or dominant, to improve genetic counseling and molecular diagnosis, and offer new perspectives of treatment. PMID- 15135430 TI - [Drug treatment of migraine in children: state of the art]. AB - Migraine, according to the criteria of the International Headache Society, occurs in about 5-10% of children. Preventive therapy includes identification of migraine precipitants, possible adjustments in lifestyle, appropriate management of acute headache, and when necessary the use of pharmacologic agents. It should be started if migraine attacks are severe or frequent. Non-pharmacologic prophylactic treatment is the modality of choice, based on relaxation or biofeedback. Despite its high incidence, only a few controlled trials have investigated the prophylactic treatment of migraine in children. Only flunarizine (5 mg/day) has been shown to be effective in two double-blind, placebo-controlled trials. Some evidence also exists that propranolol (60 mg/day) and pizotifen (0.5 1.5 mg/day) are effective. For all other drugs studied in migraine prophylaxis, the results remain vague (e.g. amitriptyline), or suggest inefficacy (e.g. clonidine, tryptophane). Most of the drugs used in the treatment of migraine in children are well tolerated. The most common adverse effects are drowsiness and bodyweight gain. PMID- 15135431 TI - [Drugs news]. PMID- 15135433 TI - [Immunization and egg allergy]. PMID- 15135434 TI - [Intestinal obstruction and geophagia in a 14- year-old child]. PMID- 15135435 TI - [Resolutive stomach volvulus after endoscopy in a 16-month-old infant]. PMID- 15135436 TI - [Early electromyogram in Guillain-Barre syndrome]. PMID- 15135437 TI - [Spiruline and malnutrition]. PMID- 15135438 TI - [Spiruline and malnutrition]. PMID- 15135439 TI - [Spiruline and malnutrition]. PMID- 15135441 TI - [Fetal digestive tract pathology, imaging findings in antenatal diagnosis]. AB - Foetal anomalies of the digestive tract are numerous and dominated by malformations, the prognosis of which may be severe. Antenatal diagnosis of these anomalies allows a better management of neonates. It is known since about 20 years, a major development because of the progress realized in the field of foetal imaging. Ultrasonography, which is the best imaging modality, can be joined by MRI if necessary. The objective of this study is to evaluate the contribution of imaging in prenatal diagnosis of digestive anomalies, throughout a review of literature. PMID- 15135442 TI - [Alarm systems for use in the treatment of primary nocturnal enuresis]. PMID- 15135443 TI - [Against influenza in children, which actions?]. AB - Each year, at any age, children are at risk of influenza illness during the epidemics. Children, especially those at school (attack rate close to 30%), have a major role in viral dissemination. Most of influenza illnesses occur in healthy children. Manifestations are typical, as in adults, in children over 5 years of age and less specific in younger children explaining misdiagnosis and underestimation in the youngest. Respiratory complications in outpatients include acute otitis media (close to 30% in children less than 3 years of age) and pulmonary disease (5 up to 10%). High fever 240 degrees C is frequent. Febrile convulsions occur in about 20% of hospitalised children aged 6 month to 5 years. Other complications (encephalitis, myositis, myocarditis, etc.) are reported. Hospitalisation rate is a severity index. In children less than 5 years of age, it is as high as 500/100,000 when risk factors exist and 100/100,000 when absent. Influenza, which is a respiratory virus, is the only one for which both a vaccine and specific treatment (anti-neuraminidases) exist. They are detailed. PMID- 15135444 TI - [Cutaneous drug reactions: pharmacogenetic news]. AB - Identifying genetic factors in severe drug eruption should improve efficacy and safety of drug prescription. The first pharmacogenetic studies concerned the polymorphism of genes encoding drug metabolism enzymes (especially for sulfamides reactions in AIDS and anticonvulsants reactions). Recent studies did not confirm the role of genotypic variation in metabolism enzymes in drug reaction. Genes controlling immune response are another research issue. Certain MHC phenotypes are linked to a higher risk of toxidermy, such as in abacavir hypersensitivity or fixed drug eruption. Indeed, severe drug eruptions seem to be linked to a specific immune response controlled by T lymphocytes. PMID- 15135445 TI - [Espace Sante Jeunes, a medical care unit for vulnerable youth]. AB - Adolescents, especially belonging to vulnerable groups, feel excluded from the health system. Indeed, they lack their own care service, between the child protection and the "social help" consultations. For five years now, the Espace sante jeunes (Young People Health Center) has tried to help them to regain control over their body, and to take care of their own health. Some patients only need to be reassured about their "normality", whereas others suffer from severe pathologies that were never treated before. PMID- 15135446 TI - [Pharmaceutical news]. PMID- 15135447 TI - Streptococcus pneumoniae in nasopharyngeal secretions of healthy children: comparison of real-time PCR and culture from STGG-transport medium. AB - Precise methods for the detection of Streptococcus pneumoniae are needed for predicting the consequences of pneumococcal conjugate vaccines on nasopharyngeal carriage. In this study, 400 nasopharyngeal swab samples from children were analyzed using a real-time pneumolysin (ply)-PCR method. The specimens were originally collected into STGG-transport medium and cultured in 1999, after which they were stored at -80 degrees C until analyzed by real-time PCR in 2001. The sensitivities of real-time PCR and culture methods were also studied by analyzing 10-fold dilutions of a pneumococcal broth culture using both methods. Of the 400 nasopharyngeal swab samples, 158 (40%) were positive in culture and 276 (69%) by real-time PCR. A minor part (4%) of the culture-positive samples remained negative by PCR. There was a trend between the quantity of genome equivalents detected by PCR and the number of colonies found in culture. When analyzing 10 fold dilutions of a pneumococcal broth culture, a higher number of genome equivalents were detected using real-time PCR than the number of colonies detected by culture. Quantitative real-time PCR provides feasible means for quantifying pneumococcal carriage. Further studies are needed to confirm that positive PCR findings really indicate the presence of viable pneumococcus in nasopharyngeal specimens. PMID- 15135448 TI - Characterization of microsatellite markers to diagnose ADPKD. AB - Autosomal dominant polycystic kidney disease (ADPKD) maps to chromosome 16p13.3 (PKD1) and to chromosome 4q21-23 (PKD2), with the likelihood of a third unmapped locus. The size and genomic complexity of the PKD1 gene make it impractical to detect mutations for prenatal diagnosis. Therefore, pedigree-based linkage analysis remains useful for diagnosis of ADPKD. Since, the complete genome sequences of chromosome 16p13.3 and 4q21-23 including PKD1 and PKD2, respectively, were reported very recently, in order to do more precise diagnosis of ADPKD, we tried to find microsatellite markers. We performed database searches of 2000 kb of genome sequence across the 16p13.3 and the 4q21-23. To determine the distribution of alleles and the degree of polymorphism of the microsatellites, genotyping experiments were performed on 48 Korean individuals. We found novel 14 microsatellite markers around ADPKD that are more polymorphic and closer to PKD1 or PKD2 than the known markers. The novel microsatellite markers were applied to diagnose ADPKD families. These novel microsatellite markers are not only useful for presymptomatic and prenatal diagnosis of ADPKD, but also applicable in the study of positional cloning, human evolution and tumor biology. PMID- 15135449 TI - Four-color multiplex 5' nuclease assay for the simultaneous detection of the factor V Leiden and the prothrombin G20210A mutations. AB - We developed a real-time multiplex four-color assay for the simultaneous detection of the factor V Leiden (FVL) and prothrombin (PT) G20210A mutations in one closed tube using a single thermocycling protocol. The assay combines the power of multiplex PCR with the specificity provided by allele-specific oligonucleotide (ASO) hybridization using the 5' nuclease assay format. Human genomic DNA is prepared from whole blood with standard procedures. A 97-bp DNA sequence of the coagulation factor V gene is co-amplified with a 111-bp DNA sequence of the coagulation factor II (PT) gene using four PCR primers. In addition, the reactions included four differentially labeled ASO probes for the specific detection of the different FVL/PT G20210A genotypes. To evaluate the assay's performance characteristics, we performed a comparison of two methods. We analyzed DNA samples from 52 individuals with known FVL/PT G20210A genotypes that were previously genotyped with an assay that combined PCR with the use of restriction fragment length polymorphisms. We found a 100% concordance between the results generated by both methodologies. We conclude that the four-color multiplex assay is specific and reproducible for the detection of the FVL/PT G20210A mutations, and it can be easily adapted for the detection of other SNPs. PMID- 15135450 TI - A pncA polymorphism to differentiate between Mycobacterium bovis and Mycobacterium tuberculosis. AB - The pyrazinamidase gene coding for the enzyme that activates the bactericidal drug pyrazinamide contains a polymorphic site that is preserved in Mycobacterium bovis. We synthesized two sets of primers, one encompassing a 180 bp fragment, and the second spanning a 726 bp fragment including the full pncA gene. Following PCR of Mycobacterium tuberculosis and M. bovis samples, it is possible to discriminate by this polymorphism between these species by digestion with Eco065 I. Digestion of the 180 bp fragment results in two fragments of 101 and 79 bp, specific for M. tuberculosis. Alternatively, digestion of the 726 bp fragment yields three fragments of 452, 165 and 109 bp for M. tuberculosis, but only two fragments of 561 and 165 bp for M. bovis. PMID- 15135451 TI - Construction and use of PCR primers from a 65 kDa mannoprotein gene for identification of C. albicans. AB - A method for detection of Candida albicans in biological samples (blood, serum, urine) was developed by the use of polymerase chain reaction (PCR) amplification of a DNA fragment of a gene coding for a 65 kDa mannoprotein of C. albicans (CaMP65). The PCR amplifies a 220 bp fragments whose specificity for C. albicans was demonstrated by Southern blot with a non-radioactive probe, leading to the differentiation from all other yeast species or human and bacterial DNA. The sensitivity of this assay was 5-10 C. albicans cells per milliliter of biological sample. PMID- 15135452 TI - Molecular dissection of the rDNA array and of the 5S rDNA gene in Meloidogyne artiellia: phylogenetic and diagnostic implications. AB - The sequence of a 13.423 nucleotide genomic fragment has been determined for the plant parasitic nematode Meloidogyne artiellia. It contains an entire rDNA cluster, the bordering intergenic regions and portions of the flanking coding regions. The sequence analysis of the rDNA repeats suggests homogeneity in M. artiellia, thus providing a further indication of the usefulness of these genes for the diagnostic identification of this species. The comparison of the secondary structures of the internal transcribed spacer 2 region in several Meloidogyne species indicates that RNA folding predictions can be used as a tool of potential diagnostic relevance. The other ribosomal gene, 5S rDNA, has been demonstrated to be functional and located near the trans-spliced leader sequences, in the same arrangement found in the distantly related nematode Caenorhabditis elegans but never in other Meloidogyne thus providing species specific markers for the identification of several Thylenchida parasitic nematodes. PMID- 15135453 TI - Detection by 5'-nuclease PCR of Shiga-toxin producing Escherichia coli O26, O55, O91, O103, O111, O113, O145 and O157:H7, associated with the world's most frequent clinical cases. AB - This paper describes 5'-nuclease PCR assays for detecting eight O-serogroups, H7 flagellar antigen and stx genes from the Shiga toxin-producing Escherichia coli (STEC) associated with the world's most frequent clinical cases. A single set of primers was used to detect the genes stx1 and stx2 in the same reaction by 5' nuclease PCR. Serotyping by 5'-nuclease PCR of STEC was based on the selection of primers and probes targeting the O-antigen gene clusters of E. coli O26, O55, O91, O111, O113, O157, the eae gene of E. coli O103, the O-island 29 of E. coli O145, and the flagellar H7 antigen gene. Results obtained on a collection of 190 strains indicate that the 5'-nuclease PCR assays used here could serve as a basis for rapid specific stx, O and H7 typing of these major pathogenic serogroups of E. coli. This work provides sensitive and specific tests for the rapid, reliable detection of the main pathogenic E. coli O-serogroups of major public health concern. PMID- 15135454 TI - A non-invasive test for sex identification in short-toed Eagle (Circaetus gallicus). AB - Knowledge of the sex of individuals in natural populations greatly facilitates evolutionary ecology, breeding systems and genetics. Therefore, the development of a simple, not stressing and objective sexing test would facilitate conservation of the Short-toed Eagle (Circaetus gallicus), an endangered Accipitridae species living mainly in southern Europe and Asia. A PCR test was used employing primers that amplify two homologous fragments of both the CHD-W gene, unique of females, and the CHD-Z, occurring in the two sexes. The analysis of the PCR products obtained from blood DNA showed a band of about 380 bp, apparently unique in all individuals. The alignment of the sequences of the two fragments revealed that CHD-W is only 9 bp longer than CHD-Z (387 vs. 378 bp) while CHD-Z lacks the restriction site for Asp700I. After digestion male PCR products showed a unique band of 378 bp while fragments belonging to females resolved into three bands (378, 280 and 107 bp). Using feathers as DNA sources, the individual patterns obtained were identical with the corresponding blood DNA samples. This sexing technique is objective and non-invasive and could be useful for verifying the sex ratio theories and improving the management. PMID- 15135455 TI - An electrophoretic tool for the genetic characterisation and delineation of lungworms. AB - In the present study, PCR-based single-strand conformation polymorphism (SSCP) analysis of the second internal transcribed spacer (ITS-2) of nuclear ribosomal DNA (rDNA) was applied to the genetic characterisation of Dictyocaulus from red deer from New Zealand and to its differentiation from species of lungworm from cattle and other hosts. Based on SSCP profiles, Dictyocaulus individuals from red deer from different geographical localities in New Zealand could be readily distinguished from those representing other lungworms examined, irrespective of low-level sequence variability in the ITS-2 (0.4-2.6%) detectable among individuals. The ITS-2 of Dictyocaulus from red deer differed in sequence by approximately 7-35% from congeners from other cervid hosts, demonstrating that this parasite is genetically distinct from other species of Dictyocaulus for which ITS-2 sequence data are presently available. The results emphasize the need for a large-scale molecular systematic study of Dictyocaulus specimens from various species of cervid and other ruminant hosts and the usefulness of mutation scanning for taxonomic, epidemiological and population genetic investigations. PMID- 15135456 TI - Polymorphism report: identification of two common single nucleotide polymorphisms in the human toll-like receptor 5 (TLR5) gene. PMID- 15135457 TI - Comparison of TaqMan and Epoch Dark Quenchers during real-time reverse transcription PCR. AB - Several biotechnology companies have recently introduced novel quencher fluors for use with dual-labeled fluorogenic hydrolysis probes. The Epoch Dark Quencher trade mark fluorochrome consists of a non-fluorescent moiety capable of absorption at higher wavelengths (400-650 nm). The aim of this study was to: (1) evaluate the feasibility of using Epoch Dark Quencher fluorochromes in real-time PCR pathogen detection assays that were previously optimized with TaqMan (TAMRA) quenching fluors, and (2) compare the sensitivity based on cycle threshold (CT) between probes containing either TaqMan or Epoch Dark Quencher fluors. Our data indicate Epoch Dark Quencher probes can be used in place of TaqMan probes and their performance was not better than traditional TaqMan (TAMRA) quenchers. Marginal differences observed between quenching fluorochromes may arise from concentration differences during probe synthesis. PMID- 15135458 TI - Anemia in the critically ill. AB - The anemia of critical illness is a distinct clinical entity with characteristics similar to that of chronic disease anemia. Several solutions to the processes of anemia, such as blunted erythropoietin production and erythropoietin response and abnormalities in iron metabolism have been developed. The transfusion of RBCs provides immediate correction of low hemoglobin levels, which may be of value in patients with life-threatening anemia. Avoidance of RBC and blood component transfusion, however, is becoming increasingly important as data of adverse clinical outcomes in critically ill patients become clearer. Although the optimal hemoglobin in critically ill patients is not determined, this organ system has a generous reserve. Short-term compensated anemia is tolerated well, while exogenous erythropoietin allows patients to achieve higher hemoglobin concentrations without exposure to transfused blood/blood components. A recent randomized trial enrolled over 1300 critically ill patients to receive either 40,000 units of exogenous erythropoietin or placebo. These authors found that patients randomized to erythropoietin received significantly less allogeneic RBC transfusions and had significantly greater increases in hemoglobin. Although no differences were found between groups in gross clinical outcomes (ie, death, renal failure, myocardial infarction), this study did not have the power to identify small differences in outcomes. This and other studies of exogenous erythropoietin therapy in critically ill patients clearly demonstrate that the bone marrow in many of these patients will respond to the administration of erythropoietin despite their illness, suggesting a blunted production of erythropoietin rather than a blunted response to erythropoietin. Exogenous erythropoietin therefore represents a therapeutic option for treating anemia in critical illness. Acute events in medicine and surgery often lead to many patients becoming anemic. Solutions to this process of anemia should be focused on preventing such events. Anemia after surgery represents an area for prevention. Blood conservation strategies can be performed with adequate results. Monk et al randomized 79 patients undergoing radical prostatectomy to preoperative autologous donation (PAD), preoperative exogenous erythropoietin therapy plus ANH immediately following induction of general anesthesia, and ANH alone. This study concluded that all three techniques resulted in similar hemostasis outcomes (eg, bleeding and transfusion rates), but ANH alone was the least expensive, and ANH plus exogenous erythropoietin and ANH alone resulted in a higher ICU hematocrit compared with PAD. Regardless of these prophylactic strategies, patients still become anemic after surgery or during critical illness. This acute event anemia usually is treated with RBC transfusion; however, autologous blood recovery (cell salvage systems) has been shown to be effective in patients with acute bleeding-related anemia, and this may reduce patients' exposure to allogeneic blood in these patients. There are no universally accepted treatment guidelines for managing anemia, and practice differs between clinicians, hospitals, regions, and countries. Transfusion medicine is evolving and incorporating many new pharmacological agents into the armamentarium of anemia and bleeding therapy. Accumulating evidence suggests that anemia in critically ill patients is common and correlated with poor outcomes. The management of anemia can improve outcomes; however, the optimal management of anemia is not performed universally. New approaches, continued research, and an understanding of anemia may result in more consistent and improved outcomes for critically ill patients. PMID- 15135459 TI - Blood transfusion practice today. AB - In trying to develop a cohesive and logical transfusion strategy, one must try to keep all of the previously mentioned information in mind. While recognizing that RBC transfusions are an integral part of care for critically ill patients, one also must recognize that there are large amounts of data that associate multiple transfusions of allogeneic blood with increased risk of morbidity and mortality. Some data exist showing causation and association, such as studies demonstrating the immunomodulatory effects of allogeneic transfusions. In addition, the blood supply is a limited resource that should not be used indiscriminately. It therefore behooves clinicians to subject each unit of blood transfused to scrutiny. In all likelihood, patients who are not actively bleeding and who are not hypovolemic probably get little to no benefit from allogeneic blood transfusions while their hemoglobin is greater than 7 g/dL. They do, however, get needless exposure to a potentially toxic substance. Although it is the authors' hope that more research will be performed to clarify the risks and benefits of blood transfusion, the authors also hope that knowledge of already published studies will continue to spread and replace the unfounded practices of the past. PMID- 15135460 TI - Physiologic aspects of anemia. AB - The most important adaptive responses from a physiological stance involved the cardiovascular system, consisting in particular of elevation of the cardiac output and its redistribution to favor the coronary and cerebral circulations, at the expense of the splanchnic vascular beds. The evidence regarding these physiological responses, especially in experimental studies that permit the control of many variables, is particularly powerful and convincing. On the other hand, there is a remarkable lack, in quality and quantity, of clinical studies addressing how normal physiological adaptive responses may be affected by a variety of diseases and conditions that often accompany and may complicate anemia, and interactions with other such compounding variables as age and different patient populations. For these reasons, it is not possible to offer guidelines on how to increase, maintain, or even to determine optimal DO2 in high risk patients and how best transfusion strategies might be used under these conditions. From the brief review of physiological principles and the strong consensus in the literature, it is evident that cardiac function must be a central consideration in decisions regarding transfusion in anemia, because of the critical role it plays in assuring adequate oxygen supply of all vital tissues. Particular attention should be paid to the possible presence of CAD or incipient or cardiac failure, as these conditions may require careful transfusions to improve DO2 at levels that may not necessitate such interventions when cardiac disease is absent. Although the cerebral circulation also serves an obligate aerobic organ unable to tolerate significant hypoxia, there is little convincing evidence to support the notion that cerebral ischemia is aggravated by anemia and that this can be prevented by improved DO2 through rapid correction of anemia. Consequently, the arguments favoring transfusions in the presence of ischemic heart disease do not appear to apply to occlusive cerebrovascular disease. Because firm evidence is lacking on the interactions of concurrent diseases and anemia in various patient populations, understanding of the physiological consequences of anemia, and of the diseases concerned, is useful but not fully sufficient to provide firm and rational guidance to transfusion practice in specific complex clinical instances. A good deal of clinical and experimental investigation is required to support fully rational and comprehensive guidelines. In the meantime, prudent and conservative management, based on awareness of risks and sound understanding of the normal and pathological physiology, must remain the guiding principle. PMID- 15135461 TI - Tissue oxygen delivery and the microcirculation. AB - In health, acute anemia is accompanied by changes in the distribution of blood flows at all of the central, regional, and microcirculatory levels. This redistribution in blood flows provides the capacity to maintain tissue oxygenation with hematocrit as low as 21%. What is not known with certainty is whether the capacity to maintain tissue oxygenation in the presence of acute anemia can be influenced significantly by concurrent disease such as sepsis and cardiac disease. The single clinical trial found an apparent survival benefit by not exposing patients with sepsis to blood transfusions until the hemoglobin concentration was less than 70 g/L. The question remains as to whether this observation was the consequence of a protective effect anemia or an injurious effect of transfusing old stored blood. PMID- 15135462 TI - Clinical consequences of anemia and red cell transfusion in the critically ill. AB - Despite the frequent use of red cell transfusions, only one large randomized trial has examined red cell administration perioperative and in the critical care setting. However, the TRICC Trial does not provide sufficient evidence to determine optimal transfusion practice in postoperative care, in critically ill children, or in patients with a myocardial infarction or acute coronary syndromes. In addition, most transfusion practice guidelines published before the completion of the TRICC Trial are now dated and need to have expert opinion informed by solid evidence in diverse clinical settings. In the next several years,several randomized trials will provide additional evidence in support of bedside decision-making. For example, two transfusion studies will be evaluating transfusion triggers, including one in premature infants and the other in critically ill children. At this juncture, high-quality clinical evidence is not yet available for many decisions related to red cell transfusions. We anticipate that risks and benefits of red cells and alternatives will be elucidated in the coming years. PMID- 15135463 TI - Blood transfusion risks in the intensive care unit. AB - This article reviews transfusion risks with particular emphasis on the critically ill. It describes the various types of noninfectious,infectious, and mild-to severe reactions that can occur in a trans-fused patient. The article describes differential diagnosis of these reactions and the handling and treatment of the patient. Diagnosis of the type of transfusion reaction by laboratory tests is detailed. Finally, the article discusses the dangers of human error with possible strategies to combat this problem using new technologies. PMID- 15135464 TI - Efficacy of red blood cell transfusion in the critically ill. AB - This article has evaluated the published data regarding the efficacy of RBC transfusions in the critically ill. Taken together, these studies generally support conservative RBC transfusion strategies in critical care to reduce the risk of transfusion-related adverse effects. The TRICC trial has established the safety ofa restrictive transfusion strategy, suggesting that physicians could minimize exposure to allogeneic RBCs by lowering their transfusion threshold. Further research will add to the generalizability of this study and explore the possible mechanism to explain why RBC transfusions do not improve outcomes in the critically ill. Additional studies will be necessary to determine the effects of RBC storage time and the presence of allogeneic leukocytes in allogeneic RBC. The following conclusions are evident: 1. RBC transfusion does not improve tissue oxygen consumption consistently in critically ill patients, either globally or at the level of the micro-circulation. 2. RBC transfusion is not associated with improvements in clinical outcome in the critically ill and may result in worse outcomes in some patients. 3. Specific factors that identify patients who will improve from RBC transfusion are difficult to identify. 4. Lack of efficacy of RBC transfusion likely is related to storage time, increased endothelial adherence of stored RBCs, nitric oxide binding by free hemoglobin in stored blood, donor leukocytes, host inflammatory response, and reduced red cell deformability. PMID- 15135465 TI - Transfusion in the cardiac patient. AB - Transfusion guidelines in patients with coexisting cardiac diseases are similar to the ones in patients without such comorbidity, in that allogeneic blood transfusions most often are indicated at hemoglobin levels of less than 6.0 g/dL and hardly ever at hemoglobin levels greater than 10 g/dL. In the hemoglobin range of 6 to 10 g/dL, signs of impaired oxygenation should serve as transfusion indications, and such signs may be reached at higher hemoglobin values than in healthy patients. An inadequate oxygenation may become manifest globally in the form of a general hemodynamic instability with a tendency to hypotension and tachycardia despite normovolemia or an oxygen extraction of greater than 50%. An inadequate oxygenation in the form of myocardial ischemia may be manifested by new ST-segment depressions of greater than 0.1 mV, new ST-segment elevations greater than 0.2 mV, or new wall motion abnormalities in transesophageal echocardiography. Institutional guidelines also should consider local logistic characteristics such as the level of knowledge of physician and nurse staff caring for patients and the level of surveillance possible justifying eventually higher hemoglobin transfusion triggers, particularly in the postoperative period. PMID- 15135466 TI - Transfusion in surgery and trauma. AB - The role of transfusion in surgery and trauma continues to evolve with our greater understanding of the true indications for and effects of transfusion. The potential adverse immune consequences and end-organ effects of blood transfusion must be weighed against the need for replacement of blood volume and oxygen carrying capacity. The techniques to conserve blood and avoid transfusion play an important role in caring for the bleeding surgical patient. The future holds great promise for the possibility of redefining the art of blood transfusion and perhaps one day replacing it entirely. PMID- 15135467 TI - Transfusion in pediatrics. AB - In reviewing the literature, the authors noted an important variation in stated and observed transfusion practice patterns among pediatric critical care practitioners, and in published guidelines on RBC transfusion. They also noted a paucity of clinical evidence with respect to RBC transfusion to critically ill children. There has been only one large randomized trial in adults, and the authors do not believe that the results from this trial should be generalized to critically ill children because of the many differences in children and their adaptive responses, and differences in disease processes. More research about anemia and RBC transfusion to critically ill children must be performed. The TRIPICU study is testing the safety of giving more or less RBC transfusion to stable critically ill children. Other studies must be done on the epidemiology and determinants of RBC transfusion in PICUs, on prevention of transfusion, and on alternatives to RBC transfusion (eg, erythropoietin). PMID- 15135468 TI - Blood conservation for critically ill patients. AB - Anemia may be the most common illness of critically ill patients. The majority of critically ill patients are anemic at admission to the intensive care unit (ICU), and hemoglobin concentrations typically decline during the first 3 days of ICU stay. Hemoglobin continues to decline for patients with sepsis and higher severity of illness. This patient population may be at particular risk of adverse consequences of anemia given the cardiovascular, respiratory, and metabolic compromise frequently encountered during critical illness. The etiology of anemia of critical illness is multifactorial, resulting from phlebotomy, gastrointestinal bleeding, coagulation disorders, blood loss from vascular procedures, renal failure, nutritional deficiencies,bone marrow suppression, and impaired erythropoietin response. PMID- 15135470 TI - Hypoxia-inducible factor 1 (HIF-1) in cancer. AB - Hypoxia is a common feature of many cancers. It contributes to local and systemic tumour progression as well as potentially compromising radiotherapy and chemotherapy. Hypoxia-inducible factor 1 (HIF-1) is an essential component in changing the transcriptional response of tumours under hypoxia. It targets the transcription of over 60 genes involved in many aspects of cancer biology including cell survival, glucose metabolism, cell invasion and angiogenesis. Over expression of HIF-1 has been associated with increased patient mortality in several cancer types including breast, stomach, cervical, endometrial and ovarian cancers. The pharmacological manipulation of HIF-1 has marked effects on tumour growth, and it could prove to be an important target for drug therapy, both in cancer and in other hypoxia-dependent disease states. PMID- 15135471 TI - Estrogen receptor alpha and beta profiling in human breast cancer. AB - BACKGROUND: The identification of a second estrogen receptor (ER-beta) has significant implications for therapeutic strategy in breast cancer management arising from the potential agonist effect of Tamoxifen at estrogen receptor sites and as such, antiestrogen therapy may be inappropriate in patients with a dominance of ER-beta. METHODS: To determine the proportion of breast cancer patients who may be so at risk, we developed a novel multiplexed RT-PCR technique to establish the relative ER-alpha and ER-beta levels in 53 primary breast cancers, 11 normal breast tissues and six cell lines. We further assessed the prognostic significance of receptor status relative to the Nottingham prognostic index (NPI). The ER-alpha and ER-beta status was also determined by immunohistochemistry using previously published and 'in-house' scoring systems. RESULTS: Using RT-PCR analysis, 46 tumours were hormone receptor positive (ER+) with 42 displaying ER-alpha predominance. Comparison with immunohistochemistry demonstrated 44/53 (ER-alpha) and 27/50 (ER-beta) concordance rates. There was no significant difference in the NPI between ER-alpha and ER-beta predominant cohorts or between ER+ and ER- cohorts. CONCLUSION: This study identifies the existence of a subgroup of ER+ patients in whom Tamoxifen therapy may be inappropriate and has significant implications for adjuvant therapy of primary breast cancer. PMID- 15135472 TI - Sentinel lymph node biopsy in patients with multifocal breast cancer. AB - BACKGROUND: Multifocal or multicentric breast cancer has been suggested as a contraindication for sentinel node biopsy (SNB). However, recent studies have demonstrated that all quadrants of the breast drain through common afferent channels to a common axillary sentinel node. This should mean that the presence of multifocal tumour should not affect the lymphatic drainage. The purpose of this study was to evaluate the feasibility and accuracy of SNB in patients with multifocal breast cancer using a peritumoural injection technique for sentinel lymph node (SN) mapping. METHODS: In the ALMANAC multicentre trial validation phase, we took SNB samples from 842 patients with node negative, invasive breast cancer with use of a blue dye and radiolabelled colloid mapping technique at the peritumoural injection site. All patients underwent standard axillary treatment after SNB. Seventy-five of the 842 patients had multifocal lesions on final histopathologic examination. The following analysis is focused on patients with multifocal lesions. RESULTS: A mean number of 2.4 SNs were identified in 71 of 75 patients (identification rate: 94.7%). Thirty-one patients had a positive SN, 40 a negative SN. Standard axillary treatment confirmed the SN to be negative in 37 of 40 patients, whereas three patients revealed positive non-sentinel lymph nodes (false-negative rate: 8.8%). Overall SN biopsy accurately predicted axillary lymph node status in 68 of 71 patients (95.8%). CONCLUSION: SNB accurately staged the axilla in multifocal breast cancer and may become an alternative to complete axillary lymph node dissection in node negative patients with multifocal breast cancer. PMID- 15135473 TI - Sentinel lymph node biopsy in male breast cancer patients. AB - The concept of sentinel node biopsy has been validated for female breast cancer patients whereas, ALND remains the standard of care for male breast cancer patients with similar tumours. We evaluated the results of SLN biopsy in male breast cancer patients with clinically negative axillae. This study included all male breast cancer patients who underwent SLN biopsy between February 1998 and October 2003. All patients had negative axillae on clinical examination. All patients underwent pre-operative lymphoscintigraphy. SLN biopsy was performed using a combination of Patent blue V and 99mTc-radiolabelled colloidal albumin injected peritumourally. Nine patients, 26-79 years of age, were included in the study. Pre-operative lymphoscinitgraphy identified SLNs in all patients. Intraoperatively, SLNs were successfully localised in all patients. The mean number of SLNs encountered was 2.4. Five patients had a positive SLN, four a negative SLN. Five patients (one with a negative SLN, four with a positive SLN) had been elected pre-operatively to undergo ALND regardless of findings on SLN biopsy. ALND confirmed the SLN to be negative in one patient (false-negative rate: 0%) and three of the four patients with positive SLN(s) had additional positive nodes in the axilla. SLN biopsy accurately predicted axillary lymph node status in these five patients. These findings compare favourably with findings reported in the literature regarding SLN biopsy in female breast cancer patients. SLN biopsy accurately staged the axilla in male breast cancer patients and should be considered for axillary staging in male breast cancer patients with clinically negative axillae. PMID- 15135474 TI - Integrin expression and survival in human breast cancer. AB - BACKGROUND: Integrin cell adhesion molecules are fundamental to numerous cellular functions including anchorage, differentiation and proliferation. Reduced expression of certain alpha and beta integrin subunits in primary breast cancer cells has been correlated with increased invasion and metastasis. Conversely, over-expression of the alpha6 subunit has been linked to poorer survival. The objective of this study was to measure the survival of a cohort with breast carcinoma in relation to integrin expression and to evaluate their potential as prognostic indicators. METHOD: Integrin expression on samples from 99 consecutive patients with breast cancer was assayed using monoclonal antibodies to the subunits alpha(1,2,3,6,V) and beta(1,3,4,5). This cohort has now been followed prospectively for almost five years allowing for early assessment of survival in relation to integrin expression. RESULTS: Whilst analysis of the data confirmed the relation of survival to proven predictors of tumour grade, tumour size and vascular invasion, statistical significance was not demonstrated with regard to both lymph node status and all integrin subunits studied. CONCLUSION: Previous research correlating certain integrin subunits with survival has not been confirmed in this study. Despite proven molecular importance in tumour cell adhesion, invasion and metastasis, integrin expression would appear not to translate clinically as independent indicators of prognosis, at least in the short-term. PMID- 15135475 TI - The effect of radiotherapy on the use of immediate breast reconstruction. AB - BACKGROUND: Immediate breast reconstruction techniques include tissue-expansion, latissimus dorsi flap with or without an implant, pedicled TRAM flap and free tissue-transfer. Adjuvant radiotherapy decreases loco-regional recurrence and increases overall survival. Radiotherapy in the presence of a tissue-expander or an implant can lead to an increased number of complications and poor cosmetic outcome. AIM OF THE STUDY: To study the relationship between radiotherapy and the choice of the immediate breast reconstruction technique in view of the increased number of breast cancer patients receiving adjuvant radiotherapy. PATIENTS AND METHODS: An audit of 121 patients who had immediate breast reconstruction over a period of 2 years was reviewed retrospectively. In March 1998, the radiotherapy protocol was revised. Forty-two patients operated on between January 1997 and March 1998 were compared to 79 patients operated on between April 1998 and June 1999. RESULTS: The percentage of patients receiving adjuvant radiotherapy increased in the second period as well as the proportion of autologous breast reconstruction. A small percentage of patients required unexpected radiotherapy after insertion of tissue expanders, due to narrow excision margins or unexpected pathology. Only two patients had tissue-expansion although radiotherapy was likely. CONCLUSION: The choice of the immediate breast reconstruction technique was satisfactory in most patients. The use of implants is best avoided in patients who may require adjuvant radiotherapy. Autologous immediate breast reconstruction, either free or pedicled flaps, is a safer choice for those patients. PMID- 15135476 TI - Preoperative serum levels of follicle stimulating hormone (FSH) and prognosis in invasive breast cancer. AB - AIMS: We investigated the association between preoperative serum levels of follicle stimulating hormone (FSH) and the prognosis in women with invasive breast cancer. METHODS: Serum levels of FSH were measured in 182 premenopausal and 581 peri- or postmenopausal women with invasive breast cancer. They were followed for a mean time of 84 months. The study endpoint was death from breast cancer (182 events). Analyses were stratified on menopausal status. RESULTS: None of the estimates showed a statistically significant result. In both pre- and postmenopausal women there was a nominally higher probability of survival with a higher FSH level. Point estimates in multivariate analysis incorporating age, tumour diameter, axillary lymph status, estrogen and progesterone receptor content and year of treatment indicated a stronger association with FSH levels in premenopausal than postmenopausal women (relative hazard 0.63 or 0.85, respectively in the highest compared with the lowest quartile). CONCLUSION: We did not find any statistically significant association between preoperative serum level of FSH and prognosis. Today, FSH is not a clinical target for intervention or a clinically useful prognostic factor and the results of clinical studies up to date can only be used for motivation of further experimental laboratory research. PMID- 15135477 TI - Contrast-enhanced MR imaging of breast lesions and effect on treatment. AB - PURPOSE: To assess the value of local staging with preoperative magnetic resonance imaging (MRI) in patients with suspect breast lesions and the effect on therapeutic approach. MATERIALS AND METHODS: Two hundred and four consecutive women with suspect breast lesions on clinical examination (CE) and/or mammography (MX) and/or ultrasound (US) underwent preoperative contrast-enhanced MRI. Detection of multifocal, multicentric and bilateral breast cancer by all three imaging modalities was evaluated. Results of preoperative breast MRI were discussed with the treating surgeons. The type of therapeutic change after preoperative MRI was marked on a questionnaire (none, additional fine needle aspiration, core biopsy, open biopsy, wider excision, mastectomy) and considered 'necessary' or 'unnecessary' using final histopathological results as gold standard. RESULTS: In 170 patients, breast cancer was diagnosed. MRI detected 96% of multifocal disease and 95% of multicentric disease, whereas MX depicted 37 and 18%, and US 41 and 9% of them, respectively. All bilateral breast cancers were seen on MRI; both MX and US detected 56%. Findings of more extensive disease and unsuspected multiple breast cancer foci identified on MRI only, changed the therapeutic approach correctly in 30.6% of breast cancer patients. Nine unnecessary wider excisions and three unnecessary FNA/core biopsies were performed because MRI overestimated the number or size of malignant lesions. CONCLUSION: Preoperative breast MRI is an important adjunct to conventional imaging in the loco-regional staging of breast cancer and a useful tool in treatment planning. PMID- 15135478 TI - Variation in lymphatic function may predispose to development of breast cancer related lymphoedema. AB - AIMS: Breast cancer-related lymphoedema (BCRL) remains a common complication of breast cancer treatment. Many features of this condition remain poorly understood, such as why only approximately 25% of women are affected after similar treatment, and the phenomenon of 'sparing', in which regions of an otherwise swollen arm, most commonly the hand, remain unaffected. This study uses dual-isotope lymphoscintigraphy, involving measurement of rate of clearance of radiolabelled protein from a subcutaneous depot and subsequent appearance in blood, to quantify alterations in lymphatic function in women with BCRL, and to further investigate differences between those in whom the hand is involved with swelling and those in whom it is spared. METHODS: Participants received a depot injection of human immunoglobulin G in the dorsum of both hands, labeled with technetium-99m on one side and indium-111 on the other. Rates of clearance from the depot and appearance in venous blood were measured at regular intervals over a 3 h period. RESULTS: A total of 18 women with a history of BCRL were studied. Significant reductions in both depot clearance and venous appearance were observed in the affected arm compared with the unaffected contralateral control. On sub-group analysis, significant differences were also observed between swollen and spared hand groups, both for the affected and unaffected contralateral arm. DISCUSSION: This study, as well as confirming impaired lymphatic function in arms affected by BCRL, also shows underlying variation in lymphatic function in the unaffected contralateral arm, between those with and without hand sparing. This raises the possibility that the risk of developing BCRL may be, in part, pre determined. PMID- 15135479 TI - Expression of prolactin receptor mRNA in oestrogen receptor positive breast cancers pre- and post-tamoxifen therapy. AB - AIM: Previous in vitro studies have shown that tamoxifen down-regulates prolactin receptors in breast cancer cells. The aim of this study was to determine whether similar changes might provide the basis for a predictive test in patients. METHODS: Biopsy specimens were obtained from 28 post-menopausal women immediately before initiation of treatment with tamoxifen (20 mg daily) and after treatment for 7 days. Prolactin receptor mRNA, determined by reverse-transcription polymerase chain reaction, was then expressed relative to 18S ribosomal RNA. RESULTS: There was good evidence for a decline in receptor expression in response to treatment with tamoxifen in the whole group (p = 0.036) but with a particularly marked decrease (>60%) in a sub-group of 11 patients. No clear correlation with tumour type or grade, or with several other markers (progesterone receptor, c-erb B-2, pS2, or Bcl-2) was apparent. CONCLUSION: Tamoxifen reduces expression of mRNA encoding the prolactin receptor in a sub group of breast tumours and might provide the basis for a predictive test for tamoxifen therapy. PMID- 15135480 TI - Tumour cell displacement after 14G breast biopsy. AB - INTRODUCTION: Seeding of biopsy needle tracks with viable malignant cells was an initial concern with all diagnostic breast needle procedures, including 14G automated needle biopsy. In an attempt to further evaluate this phenomenon, we have addressed the following questions: (1) are the tracks left by the needle biopsy procedure detectable in the surgical excision specimen? (2) Are displaced tumour cells visible along the needle tracks? (3) Is it possible to identify and excise the entire needle track for thorough histopathologic evaluation? METHODS: Surgical biopsy specimens of patients previously diagnosed with cancer on stereotactic 14G-needle biopsy were studied to find needle tracks. These are characterised by hemosiderin, scar tissue, foreign body giant cell reaction, fat necrosis, or organizing haemorrhage. Occurrence of displaced tumour cells or groups of cells along the tracks was registered. RESULTS: Needle tracks were found in 22/64 excision specimens of patients who underwent 14G-needle biopsy and surgery on the same day. Tumour-cell displacement along the needle track was seen in 11/22 cases (50%). In a prospective study, an attempt was made to excise the entire needle track in 13 consecutive cancer cases after 14G biopsy. Median time interval between core biopsy and first surgical excision was 21 days (range 7 35). Needle tracks could be visualised in 11/13 cases; displaced cells were seen in seven. CONCLUSION: Needle tracks can be found in the excision specimens of patients who previously underwent 14G biopsy, and displaced tumour cells can be recognised. Excision of the entire needle track is neither feasible, nor advisable as a routine measure, since radiotherapy is advised for both in situ and invasive carcinoma after conservative surgery. PMID- 15135481 TI - Seroma prevention following axillary dissection in patients with breast cancer by using ultrasound scissors: a prospective clinical study. AB - AIMS: Seroma formation following axillary dissection is a common complication of breast surgery. The aims of this study were (1) to analyse the risk factors of seroma formation, and (2) to evaluate the role of ultrasound scissors in performing axillary dissection in patients with primary breast cancer undergoing mastectomy and breast-conserving surgery. METHODS: Ninety-two women (median age 55 years, range 33-73 years) requiring surgery for known unilateral primary breast cancer (pT1a=1, pT1b=20, pT1c=43, pT2=25, pT3=3) were prospectively randomised to undergo axillary dissection by either using (Group A, 45 patients) or not using (Group B, 47 patients) ultrasound scissors (US). Thirty-eight (41.3%) patients underwent modified radical mastectomy, while 54 (58.7%) underwent breast-conserving surgery. RESULTS: Twenty-eight (30.4%) patients (Group A=9 out of 45, 20%; Group B=19 out of 47, 42%; P=NS) developed a wound seroma. Multivariate analysis using a logistic regression model showed that surgical procedure (RR=8.9; 95% CI: 3.2-25.3), total amount of drainage (RR=7.8; 95% CI: 2.8-22.0), and size of the tumour (RR=6.0; 95% CI: 2.2-16.5) independently correlated with seroma formation. The logistic regression function (RR=19.4; 95% CI: 6-62) correctly allocated 75 out of 92 (81.5%) patients. CONCLUSIONS: Size of the tumour, and total amount of drainage represent the principal factors of seroma formation following axillary dissection in patients undergoing surgery for breast cancer. Although the use of ultrasound cutting devices may reduce the risk of seroma formation, further studies are need to verify the real impact on long-term morbidity of such technique. PMID- 15135482 TI - Germline mutational analysis of CDH1 and pathologic features in familial cancer syndrome with diffuse gastric cancer/breast cancer proband in a Chinese family. AB - AIMS: Hereditary non-polyposis colorectal cancer, thyroid medullary carcinoma, breast/ovarian cancer and gastric cancer/breast cancer syndrome are encountered in surgery. Some gastric cancer/breast cancer syndrome may be the result of a CDH1 germline mutation. This is the first report of CDH1 germline mutations gastric cancer/breast cancer syndrome in Chinese patients. METHODS: Peripheral blood from the proband, as well as, her first and second degree relatives was collected and CDH1 gene exon 1-16 mutations were screened. E-cadherin/beta catenin proteins expression and histopathologic features were examined on gastric cancer/breast cancer tissues from the proband. RESULTS: A C-->T nucleotide substitution at exon 13 (mRNA 2200 locus, Accession number NM-004360) was found. This was a transition from GCC-->GCT in DNA sequence (Ala154Ala). Diffuse-type gastric cancer and infiltrating ductal breast carcinoma were present. Both tumours preserved E-cadherin/beta-catenin expression immunohistochemically. CONCLUSIONS: Familial cancer syndrome with diffuse-type gastric cancer/breast cancer proband in Chinese has a propensity of early onset during lifespan. No truncating or splice-site CDH1 mutations had been identified in this family. A silent nucleotide variation in exon 13 of the CDH1 gene may contribute to some forms of cancer susceptibility. PMID- 15135483 TI - A phase II study of G17DT in gastric carcinoma. AB - PURPOSE: G17DT is a gastrin immunogen, raising antibodies that blockade gastrin stimulated growth. The aim of the study was to characterise antibody response and assess safety and tolerability of G17DT given to patients with gastric cancer. EXPERIMENTAL DESIGN: G17DT was administered to 52 patients with gastric adenocarcinoma at weeks 0, 2 and 6 by intramuscular injection at doses of 10, 100 and 250 microg. Antibody levels were measured by an ELISA assay. A radioligand displacement assay determined the ability of G17DT-immunised patients' sera to inhibit binding of 125IG17 to cholecystokinin (CCK)-2 receptors. RESULTS: By week 12 of the study, 6/12 evaluable stage I-III patients achieved an antibody response in the 10 microg group, 7/11 in the 100 microg group, and 11/12 in the 250 microg group. Stage IV patients dosed at 250 microg achieved a similar response rate to stage I-III patients dosed at 10 or 100 microg. G17DT was well tolerated in 47/52 patients. Two patients suffered significant adverse reactions including injection site pain and abscess. G17DT antibodies displaced iodinated gastrin from CCK-2 receptors, with the level of displacement correlating with antibody titre. CONCLUSIONS: G17DT immunisation is a well-tolerated method of raising functional antibodies to 17 amino acid gastrin forms in patients with gastric carcinomas. PMID- 15135484 TI - Effects of neoadjuvant radio-chemotherapy on 18F-FDG-PET in esophageal carcinoma. AB - AIM: To investigate whether results of [F-18]-fluorodeoxy-d-glucose (FDG) positron emission tomography (PET) of esophageal cancer (EC) before and after neoadjuvant radio-chemotherapy correlate with histopathology after esophageal resection. METHODS: Twenty consecutive patients with EC without distant metastases were examined twice with 18F-FDG-PET during primary staging and after neoadjuvant radio-chemotherapy. FDG standardised uptake values (SUV) were correlated with the histopathological findings (percentage of viable tumour cells, tumour regression grade 1-5). RESULTS: Regression analysis revealed a slight (not significant) positive correlation between SUV(pre) (R=0.41, p=0.08) and SUV(post) (R=0.37, p=0.11) and the percentage of viable tumour cells in the resectate. Although all patients showed a significant decrease in SUV after radio chemotherapy (p < 0.01) the percentual decrease of the SUV after therapy (DeltaSUV%) did not significantly differ between the TRG-groups. In 12 of 20 patients (60%), therapy-induced esophagitis was detected in post-therapeutic PET images. CONCLUSION: In EC, a higher pre-therapeutic SUV might be correlated with a higher fraction of vital tumour cells remaining after radio-chemotherapy. Applying the neoadjuvant therapy protocol and the study design used in this examination, there is no correlation between decrease in SUV and histopathology. PMID- 15135485 TI - Intraoperative radiofrequency ablation (RFA) for irresectable liver malignancies. AB - AIMS: To evaluate the outcome of patients who received radiofrequency ablation (RFA) at open laparotomy in patients with irresectable liver malignancies. METHODS: Twenty-six consecutive patients who underwent explorative laparotomy and were found to be irresectable or who had been assessed not suitable for either resection or percutaneous RFA received intraoperative RFA. An expandable electrode (RITA Medical systems) was used. Follow-up comprised CT-scans in 6-12 week intervals. RESULTS: Patients' age ranged from 35 to 72 years (median 61). A variety of pathologies were treated. In 26 patients, 88 hepatic lesions were treated: 32 with resection and 56 with RFA. The mean diameter was 4.0+/-2.6 cm with a maximum of 10 cm. In 22 patients with hepatic metastases (18 colorectal, one leiomyosarcoma, one endometrium carcinoma, one renal cell carcinoma, one malignant phaeochromocytoma) 74 lesions (median 3.4 per patient) were treated (25 by resection, 49 by RFA). Eleven patients received simultaneous resection and RFA: resection of anatomical segments in six and atypical resection in seven patients. Procedure related complication rate was 19.2%. The mean follow-up was 14.6+/-9.2 months (2-36 months). Three patients developed recurrence at the site of previous RFA indicating incomplete ablation. The overall local control rate after one year was 92 and 90.9% for patients with colorectal liver metastases, respectively. Seventeen patients (65.4%) suffered from tumour progress. In 14 patients (53.9%) tumour occurred at new hepatic localisations and in five patients extrahepatic tumour relapse was diagnosed. Twelve patients have died so far (median survival 18 months, range 4-27). Nineteen patients had either completed a follow-up of at least 12 months or died within this period, resulting in an one year survival rate of 79% (80% for liver metastases). CONCLUSION: Intraoperative RFA is a valuable tool in liver surgery which extends the surgical spectrum in cases of irresectable malignancies. PMID- 15135486 TI - Mortality associated with pheochromocytoma in a large Swedish cohort. AB - BACKGROUND: The aim of the present study was to report the risk of death in a national cohort of patients with aPC (adrenal PC) and their risk of developing a second tumour. METHODS: Using the National Cancer Registry, 481 patients (222 men and 259 women) with aPC in Sweden (1958-1997) were identified. Autopsy-based diagnoses were excluded. As control group the entire Swedish population was used and the risk of death in patients after diagnosis of aPC was compared with the normal risk taking age, sex and calendar year into account. The risk for a second tumour disease after diagnosis of aPC was also calculated. RESULTS: Patients with aPC had an increased tumour-related mortality after diagnosis of aPC. For both men and women this mortality was four times higher than for controls. Liver/biliary tract and CNS tumours in men; and malignant melanoma and uterine cervical cancer in women were significantly over-represented in the cohort of patients with aPC. CONCLUSION: Patients with aPC run an increased risk of developing additional cancers. Surveillance strategies may thus be necessary for these patients. PMID- 15135487 TI - Clinicopathologic significance of plasma matrix metalloproteinase-2 and -9 levels in patients with undifferentiated nasopharyngeal carcinoma. AB - Increased in plasma pro-MMP2 and pro-MMP9 levels in patients with advanced stage NPC were observed. Plasma pro-MMP2 is a significant independent prognostic marker for undifferentiated NPC. AIM: Upregulation of matrix metalloproteinase-2 and -9 (MMP-2 and MMP-9) expression is observed in many cancers and high level of these proteins are found in peripheral blood of many cancer patients. In this study, we aimed at evaluating the plasma pro-MMP2 and pro-MMP9 pro-enzymes (pro-MMP2 and pro-MMP9) levels and their clinical significances in patients with undifferentiated nasopharyngeal carcinoma (NPC). METHODS: The plasma pro-MMP2 and pro-MMP9 levels were measured in 40 NPC patients and 40 normal individuals by enzyme linked immunosorbant assay. RESULTS: By using the Cox-regression model, a high pro-MMP2 level was found to be significantly correlated with poorer survival. Patients with plasma pro-MMP2 below 650 ng/ml had higher 5-year survival rate of 89%, compared with 50% for patients with plasma pro-MMP2 above 650 ng/ml. CONCLUSIONS: A high level of plasma pro-MMP2 was associated with poor survival of NPC patients independent of sex, age and stage. PMID- 15135488 TI - Prognostic factors in soft tissue leiomyosarcoma of the extremities: a retrospective analysis of 42 cases. AB - AIMS: Soft tissue leiomyosarcomas (LMS) are rare tumours which are associated with a poor prognosis. The goals of the present study were to describe the clinico-pathological and prognostic features of 42 patients affected by soft tissue LMS of the extremities. METHODS: The following clinical and pathological parameters were analysed: age, sex, site, size, depth, previous surgical procedures, stage, histological type, nuclear atypia, grade, mitotic activity, necrosis, surgical margins, therapy. Disease-free survival rates were calculated. RESULTS: The overall 2-year and 5-year disease free survival rates were 42.3% and 32.6%, respectively. By univariate analysis, tumour size (< or =10 cm vs. >10 cm; p = 0.01), average mitotic rate (< or =19/10 HPF vs. >19/10 HPF; p = 0.05), type of excision (wide vs. marginal or intralesional; p = 0.001) and adjuvant radiotherapy (none vs. brachytherapy+external beam radiotherapy vs. external beam radiotherapy; p = 0.02) were significantly correlated with disease progression. By multivariate analysis, the only factor that was found to be an independent predictor of disease relapse was type of excision (p = 0.001). CONCLUSIONS: Large tumour size and high mitotic rate resulted adverse prognostic factors. Adjuvant radiation therapy, in combination with wide surgical excision, allowed the best chance of cure. PMID- 15135489 TI - Cytoreductive surgery plus intraperitoneal hyperthermic perfusion is an effective treatment for metastasized malignant mixed mesodermal tumours (MMMT)--report of six cases. AB - BACKGROUND: Malignant mixed mesodermal tumours (MMMT) of the female genital tract are rare and heterogeneous malignancies that impart grim prognosis. These tumours are characterized by an admixture of malignant epithelial and stromal elements comprising carcinomatous and sarcomatous neoplastic cells. Thus far, almost 350 cases of MMMT have been recorded in the international medical literature. Due to its rarity, there is no agreement on the best treatment strategy in women with metastasized MMMT. METHODS: Six women (mean age 59 years) with metastasized MMMT defined to the peritoneal cavity have been treated by cytoreductive surgery plus hyperthermic peritoneal perfusion plus postoperative adjuvant chemotherapy. All patients have been pre-treated by surgery for primary tumour and one by systemic chemotherapy. As cytostatics for hyperthermic peritoneal perfusion, we have used Mitomycin in a dosage of 18 mg/m2 plus Melphalan in a dosage of 25 mg/m2. As adjuvant treatment CDDP 40 mg/m2/dl, Mitomycin 7 mg/md2/dl and Ifosfamid 100 mg/kg 24 h/dl was applicated via intraaortic catheter three times with a treatment free interval of 3 weeks. RESULTS: A complete cytoreduction without remnant tumour formations in the peritoneal cavity could be carried out in all six patients. The postoperative course was uneventful in all cases except for one where a spontaneous small bowel perforation and prolonged gall secretion had to be treated by re-operation. One patient died 4 months later by pneumonia without evidence of disease. Four patients are without evidence of disease after 2, 4, 14 and 19 months, whereas one patient developed liver metastases after 9 months still treated by systemic chemotherapy. CONCLUSION: Complete cytoreduction plus hyperthermic peritoneal perfusion plus adjuvant chemotherapy seems to be an effective treatment for recurrent or metastasized MMMT. Further studies have to define the value of this new treatment strategy for this rare tumour entity. PMID- 15135490 TI - Environmental impact of toxic pollutants. PMID- 15135491 TI - Acoustic cell filter: a proven cell retention technology for perfusion of animal cell cultures. AB - This article is a review highlighting the application of the acoustic filter as a reliable cell retention device during the long-term perfusion of animal cell cultures. Critical operating parameters such as duty cycle, perfusion and re circulation flow rates, acoustic power and backflush frequency are discussed with regard to influence on the separation efficiency and optimal operating ranges have been identified. Perfusion data gathered from the literature have been complemented with original data from a series of perfusion experiments carried out in the context of industrial projects for industrially relevant cell lines including NS0, HEK-293, SP2-derived hybridoma and insect cells in different serum supplemented and serum-free media at different perfusion rates and acoustic chamber volumes. Finally, scale-up potential of the acoustic filter for large scale industrial applications is discussed. PMID- 15135492 TI - Treatment failure in typhoid fever with ciprofloxacin susceptible Salmonella enterica serotype Typhi. AB - Fluoroquinolones are considered the most effective drugs for the treatment of typhoid fever and hence are widely used in the empiric treatment of acute undifferentiated febrile illnesses in India. Recent reports of Salmonella enterica serotype Typhi (S. Typhi) strains with increasing minimum inhibitory concentration (MIC) of ciprofloxacin have raised the fear of potential treatment failures. In this case series of 109 consecutive patients hospitalized with typhoid fever (S. Typhi grown from blood), we documented clinical failure (fever persisting >6 days) in 25 of 46 (54.3%) adults who could be evaluated. Among these, eight (17.4%) had microbiological failure (S. Typhi recovered from blood after 6 days of ciprofloxacin therapy) despite adequate serum ciprofloxacin levels, and all required alternative drugs for treatment. These 8 S. Typhi strains, although susceptible to ciprofloxacin (MIC < 1 microg/mL) (NCCLS 2000), had MICs (median MIC 0.5 microg/mL) that were increased 15-fold compared to S. Typhi strains from patients with typhoid fever seen at our center in 1995 (median MIC 0.032 microg/mL), and were nalidixic acid resistant S. Typhi (NARST) (MIC > or =32 microg/mL). The poor treatment outcomes with ciprofloxacin therapy in patients infected with NARST strains that exhibit an increased ciprofloxacin MIC call for a need to revise the ciprofloxacin breakpoints for S. Typhi. PMID- 15135493 TI - Comparison of the double-disk, combined disk, and Etest methods for detecting metallo-beta-lactamases in gram-negative bacilli. AB - Three phenotypic methods for detecting metallo-beta-lactamases (MBLs) in gram negative bacilli were tested on 60 MBL producers and 155 MBL nonproducers. The 2 mercaptopropionic acid double-disk potentiation method using ceftazidime and cefepime with and without clavulanate was found to be most sensitive (100%). The combined disk method depends on comparing the zones given by disks containing a beta-lactam with and without ethylenediaminetetraacetic acid. Various drug combinations were tested, and the best results were obtained with imipenem for Pseudomonas species and Acinetobacter baumannii, ceftazidime-clavulanate for Klebsiella pneumoniae, and cefepime-clavulanate for Enterobacter cloacae and Citrobacter freundii. The overall sensitivity of the combined disk method was 86.7%. The Etest (AB BIODISK, Solna, Sweden) method using imipenem- ethylenediaminetetraacetic acid strips detected imipenem-resistant MBL-producing isolates only, resulting in a sensitivity of 36.7%. Our data suggest that the double-disk and combined disk tests are acceptable methods for MBL detection in both imipenem-resistant and imipenem-susceptible isolates. PMID- 15135494 TI - Comparison of the BDProbeTec ET system with the roche COBAS AMPLICOR System for detection of Mycobacterium tuberculosis complex in the respiratory and pleural fluid specimens. AB - The new BDProbeTec ET System (BDET; BD Biosciences, Sparks, MD) was compared with the Roche COBAS AMPLICOR System (CAS) and culture for Mycobacterium tuberculosis (MTB). A total of 253 specimens (152 respiratory and 101 pleural fluid specimens) collected from 240 patients were tested in parallel with the 3 assays. After resolving the discrepancies, the sensitivity, specificity, and positive and negative predictive values of the BDET for detecting MTB was 76.9%, 93.7%, 71.4%, and 95.2% for the respiratory specimens and 88.9%, 92.4%, 53.3%, and 98.8% for the pleural fluid specimens, respectively. The corresponding values of the CAS were 69.2%, 100%, 100%, and 94% for the respiratory specimens and 33.3%, 100%, 100%, and 93.9% for the pleural fluid specimens, respectively. No significant differences in sensitivities were observed between the results of both assays for the respiratory specimens. However, statistically significant differences in sensitivities were found between the BDET and CAS for the pleural fluid specimens (P =.02). Although the BDET was less specific than the CAS (P =.007), the BDET has an excellent sensitivity for detecting MTB in the pleural fluid specimens. Considering the low sensitivity of other available tests, the BDET can be a useful diagnostic tool for excluding MTB, particularly in the pleural fluid specimens. PMID- 15135495 TI - Mycobacterium celatum, an emerging pathogen and cause of false positive amplified Mycobacterium tuberculosis direct test. AB - Mycobacterium celatum is a recently described organism. Herein we describe a case of M. celatum lymphadenitis in an immunocompetent child and the first reported case of a M. celatum infection (lung abscess) in a transplant recipient. A literature review identified 19 other cases of M. celatum infection. Fifteen occurred in patients with the acquired immunodeficiency syndrome. Of these, nine were disseminated and six were localized (primarily to the lungs). The remaining 4 patients were immunocompetent and had localized infection (pneumonitis or lymphadenitis). Diagnosis of M. celatum infection can be challenging as M. celatum can cause false-positive results with the current version of the Amplified Mycobacterium tuberculosis Direct Test (Gen-Probe, San Diego, CA.). Definitive identification is available by DNA sequencing or high-performance liquid chromatography. M. celatum can cause infection in immunocompromised and immunocompetent hosts. PMID- 15135496 TI - Detection and differentiation of Plasmodium species by polymerase chain reaction and colorimetric detection in blood samples of patients with suspected malaria. AB - Polymerase chain reaction (PCR) is now recognized as a sensitive and specific method for detecting Plasmodium species in blood. In this study, we tested 279 blood samples, from patients with suspected malaria, by a PCR assay utilizing species-specific colorimetric detection, and compared the results to light microscopy. Overall, both assays were in agreement for 270 of the 279 specimens. P. vivax was detected in 131 (47.0%) specimens, P. falciparum in 64 (22.9%) specimens, P. ovale in 6 (2.1%) specimens, and P. malariae in 5 (1.8%) specimens. Both P. falciparum and P. vivax were detected in a further 10 (3.6%) specimens, and 54 (19.3%) specimens were negative by both assays. In the remaining nine specimens, microscopy either failed to detect the parasite or incorrectly identified the species present. In summary, the sensitivity, specificity and simplicity of the PCR assay makes it particularly suitable for use in a diagnostic laboratory. PMID- 15135497 TI - In vitro selection of resistance in haemophilus influenzae by 4 quinolones and 5 beta-lactams. AB - We tested abilities of ciprofloxacin, levofloxacin, gatifloxacin, moxifloxacin, amoxicillin, amoxicillin/clavulanate, cefixime, cefpodoxime, and cefdinir to select resistant mutants in 5 beta-lactamase positive and 5 beta-lactamase negative Haemophilus influenzae strains by single and multistep methodology. In multistep tests, amoxicillin, amoxicillin/clavulanate and cefpodoxime exposure did not cause >4-fold minimum inhibitory concentration (MIC) increase after 50 days. One mutant selected by cefdinir had one amino acid substitution (Gly490Glu) in PBP3 and became resistant to cefdinir. Cefixime exposure caused 8-fold MIC increase in 1 strain with TEM but the mutant remained cefixime susceptible and had no alteration in PBP3 or TEM. Among 10 strains tested, ciprofloxacin, moxifloxacin, gatifloxacin, levofloxacin caused >4-fold MIC increase in 6, 6, 5, and 2 strain, respectively. Despite the increases in quinolone MICs, none of the mutants became resistant to quinolones by established criteria. Quinolone selected mutants had quindone resistance-determining region (QRDR) alterations in GyrA, GyrB, ParC, ParE. Four quinolone mutants had no QRDR alterations. Among beta-lactams cefdinir and cefixime selected one mutant each with higher MICs however amoxicillin, amoxicillin/clavulanate, and cefpodoxime exposure did not select resistant mutants. PMID- 15135498 TI - Prevalence of extended spectrum beta-lactamase producing Escherichia coli and Klebsiella isolates in a large community teaching hospital in Connecticut. AB - The prevalence of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (ESC) and Klebsiella (KS) among consecutive non-urine isolates were evaluated. Twenty-four of 392 isolates produced ESBLs. Among these half were from respiratory sites and all were susceptible to meropenem. Pulse field-gel electrophoresis (PFGE) showed 12 clonally distinct ESBLs and iso-electric focusing (IEF) revealed that most (83%) produced multiple enzymes. PMID- 15135499 TI - In vitro killing of parenteral beta-lactams against standard and high inocula of extended-spectrum beta-lactamase and non-ESBL producing Klebsiella pneumoniae. AB - Minimum inhibitory concentrations and time-kill curves were performed against 8 Klebsiella pneumoniae (4 non-extended-spectrum beta-lactamase[ESBL] and 4 ESBL) for piperacillin/tazobactam (40/5 microg/mL), cefepime (20 microg/mL), and meropenem (4 microg/mL) by using a standard and high inocula. Imipenem was evaluated only at the standard inoculum for the non-ESBL and ESBL isolates. Samples were withdrawn at 7 predetermined time-points over 24 hours and plated on trypticase soy agar plates. Minimum inhibitory concentrations were: piperacillin/tazobactam 4 to 8 microg/mL (ESBL and non-ESBL), cefepime 1 to 2 microg/mL (ESBL) and 0.06 to 0.125 microg/mL (non-ESBL), imipenem 0.125 to 0.25 microg/mL (ESBL and non-ESBL), and meropenem 0.03 to 0.06 microg/mL (ESBL and non ESBL). Each antibiotic reached and maintained bactericidal killing (> or =3 log killing) for 24 hours against all non-ESBL isolates for both the standard and high inoculum. Cefepime, imipenem, and meropenem showed the same bactericidal activity against each ESBL isolate at the standard inoculum, whereas piperacillin/tazobactam showed bactericidal killing against only 1 ESBL isolate. At the high inoculum, cefepime and piperacillin/tazobactam were unable to maintain bactericidal activity against any of the ESBL isolates. Only meropenem was able to maintain bactericidal killing over 24 hours against the ESBL isolates at the high inoculum. In summary, meropenem and imipenem maintained bactericidal activity against non-ESBL and ESBL K. pneumoniae irrespective of the inoculum size. While piperacillin/tazobactam and cefepime are bactericidal against non ESBL K. pneumoniae, their activity against ESBL K. pneumoniae is limited and based on the size of the inoculum. Until more data are available, piperacillin/tazobactam and cefepime should not be used for the treatment of ESBL K. pneumoniae. PMID- 15135500 TI - Epidemiology of macrolide and/or lincosamide resistant Streptococcus pneumoniae clinical isolates with ribosomal mutations. AB - Twenty macrolide and/or lincosamide resistant Streptococcus pneumoniae clinical isolates from various sources with 50S ribosomal mutations were identified. Mutations were identified in the 23S rDNA with substitutions at A2058, A2059, or C2611 and in L4 or L22 ribosomal protein genes. Fourteen were A2059G substitutions, one was A2058G, two were C2611T, two had an altered L4 and one isolate contained an altered L22 gene. Susceptibility testing with erythromycin, josamycin, clindamycin, and two ketolides including cethromycin was performed. The L4 mutants had the amino acid changes of (69)GTG(71) to (69)TPS(71). The isolate with the L22 mutation contained an 18 base pair tandem duplication/insertion at the 3' end of the gene. 50s ribosomal mutations are the least frequent mechanism of S. pneumoniae resistance, occurring at an extremely low frequency and are identified only by genome sequence data. PMID- 15135501 TI - Antimicrobial susceptibility patterns of Streptococcus pneumoniae in Mexico. AB - The susceptibility to 14 beta-lactam and non-beta-lactam antimicrobial agents was evaluated for Streptococcus pneumoniae from patients with community-acquired respiratory infections in a Mexican medical center. Three hundred fifteen pneumococcal isolates obtained from patients between 1995 and 2001 were tested by the broth microdilution test. Fifty-two percent of the isolates were nonsusceptible to penicillin (minimal inhibitory concentration, >0.06 microg/mL). Penicillin-nonsusceptible isolates were more likely to exhibit resistance to cephalosporins, macrolides, ciprofloxacin, trimethoprim/sulfamethoxazole, chloramphenicol, and tetracycline when compared to penicillin-susceptible isolates. Ninety-three percent of the penicillin-nonsusceptible isolates were resistant to at least one other class of antimicrobials, in contrast to only 47% of the penicillin-susceptible strains (p < 0.0001). More than 90% of the tested isolates were susceptible to amoxicillin/clavulanate, ceftriaxone, levofloxacin, and gatifloxacin. Reduced susceptibility to penicillin was considered to be a reliable marker for the higher probability of multidrug resistance, thus requiring in vitro tests to guide chemotherapy or the choices of parenteral extended spectrum cephalosporins or newer respiratory quinolones. PMID- 15135502 TI - Automated sample preparation for the TRUGENE HIV-1 genotyping kit using the MagNA pure LC instrument. AB - Use of the MagNA Pure LC Total Nucleic Acid Isolation Kit-Large Volume in conjunction with the TRUGENE HIV-1 Genotyping Kit yielded consistent, interpretable sequence data from 1-mL plasma preparations containing HIV-1 RNA concentrations of > or =400 copies/mL. This finding was confirmed in 18 of 24 low titer clinical specimens. PMID- 15135503 TI - Antimicrobial spectrum and activity of NVP PDF-713, a novel peptide deformylase inhibitor, tested against 1,837 recent Gram-positive clinical isolates. AB - Continued emergence of antimicrobial resistances among gram-positive pathogens requires further development of compounds with novel modes of action. The peptide deformylase inhibitor NVP PDF-713 was tested against 1,837 recent strains of Gram positive organisms. All NVP PDF-713 MICs were at < or = 4microg/mL except for 6 enterococci (0.3% of strains overall). NVP PDF-713 MIC(90) results were: Staphylococcus aureus, beta-haemolytic and viridans group streptococci and Streptococcus bovis at 1 microg/ml; coagulase-negative staphylococci, Streptococcus pneumoniae, and Listeria spp. at 2 microg/mL; and the enterococci at 4 microg/mL. NVP PDF-713 appears to be a promising new agent worthy of continued in vivo development. PMID- 15135504 TI - Effect of antibiotic sequence on combination regimens against Pseudomonas aeruginosa in a multiple-dose, in vitro infection model. AB - The goal of this study was to investigate the effect of antibiotic sequence on combination regimens against Pseudomonas aeruginosa in an in vitro infection model. Ceftazidime plus ciprofloxacin and ceftazidime plus tobramycin were dosed every 12 h for 48 h using simultaneous or staggered administration. Simultaneous dosing and ceftazidime followed by ciprofloxacin or tobramycin were significantly more active at both 24 h (p = 0.03) and 48 h (p < 0.0001) than ciprofloxacin or tobramycin followed by ceftazidime. Final bacterial kill was sixfold greater with the former regimens. This study showed that antibiotic sequence had a significant and class dependent effect on antibacterial response. The clinical relevance of these observations warrants further investigations in animal models. PMID- 15135505 TI - Campylobacter infection in 682 bulgarian patients with acute enterocolitis, inflammatory bowel disease, and other chronic intestinal diseases. AB - The aim of the study was to assess Campylobacter infections in 309 patients with acute enterocolitis, 272 patients with relapses of chronic enterocolitis, 70 patients with inflammatory bowel disease (involving Crohn's disease and ulcerative colitis) and 31 patients with other chronic intestinal illnesses. Isolation and identification were performed conventionally. Limited agar dilution method was used for susceptibility testing of the strains. Campylobacter species were isolated in patients with acute enterocolitis (7.8%), chronic enterocolitis (6.2%), Crohn's disease (6.2%), ulcerative colitis (3.7%), and irritable bowel syndrome (8.3%). Hippurate-positive Campylobacter jejuni isolates accounted for 62.2% of Campylobacter strains. One tetracycline resistant Campylobacter upsaliensis isolate was detected from a girl with acute enterocolitis. Resistance rates to erythromycin (31.1%) and clarithromycin (22.2%) were high, whereas those to amoxicillin/clavulanate (4.4%), ampicillin/sulbactam (13.3%), tetracycline (24.4%) and ciprofloxacin (22.2%) were relatively low. Resistance to erythromycin and either tetracycline or ciprofloxacin was detected in 8.9% and 6.7%. The involvement of Campylobacter infection in relapses of chronic intestinal disorders and the susceptibility patterns of the strains strongly emphasize the role of Campylobacter as a cause of infection in this group of patients. PMID- 15135506 TI - Been to the library lately? Veterinary Microbiology hits a century. PMID- 15135507 TI - Serotypes and virulence genes of bovine Shigatoxigenic Escherichia coli (STEC) isolated from a feedlot in Argentina. AB - Grazing-fed cattle were previously demonstrated to be reservoir of non-O157 Shigatoxigenic Escherichia coli (STEC) serotypes in Argentina. The acid resistance of some STEC strains makes it reasonable to assume the presence in feedlot of particular STEC serotypes. Fifty-nine animals were sampled every 2 weeks during 6 months by rectal swabs. Twenty-seven of 59 animals (45.8%) were shown to be Stx2(+); 3/59 (5.1%) carried Stx1(+) and 7/59 (11.9%) were Stx1(+) Stx2(+). Among 44 STEC isolates, 31 isolates were associated to 10 O serogroups (O2, O15, O25, O103, O145, O146, O157, O171, O174, O175) and 13 were considered non-typable (NT). Six H antigens (H2, H7, H8, H19, H21, H25) were distributed in 21 isolates whereas 23 were non-mobile (H-). Seventeen of 44 strains (38.6%) were eaeA(+) and 14 (31.8%) harbored the 60MDa plasmid. The megaplasmid (Mp) and eaeA gene were simultaneously found in a limited number of serotypes belonging to the enterohaemorrhagic E. coli (EHEC). E. coli O157:H7 strains, isolated from four (6.8%) animals, corresponded to the Stx2(+), eaeA(+), Mp(+) pattern. Three O157:H7 strains belonged to phage type 4 and the other strain was atypical. Many serotypes isolated from grain-fed cattle (O2:H25, O15:H21, O25:H19, O145:H-, O146:H-, O146:H21, O157:H7, O175:H8) also differed from those isolated by us previously from grazing animals. The serotypes O15:H21, O25:H19 and O175:H8 had not been identified at present as belonging to STEC. This work provides new data for the understanding of the ecology of STEC in grain-fed cattle and confirms that cattle are an important reservoir of STEC. PMID- 15135508 TI - Diagnostic multiplex PCR for toxin genotyping of Clostridium perfringens isolates. AB - In this study we provide a protocol for genotyping Clostridium perfringens with a new multiplex PCR. This PCR enables reliable and specific detection of the toxin genes cpa, cpb, etx, iap, cpe and cpb2 from heat lysed bacterial suspensions. The efficiency of the protocol was demonstrated by typing C. perfringens reference strains and isolates from veterinary bacteriological routine diagnostic specimens. PMID- 15135509 TI - Both ApxI and ApxII of Actinobacillus pleuropneumoniae serotype 1 are necessary for full virulence. AB - Most serotypes of A. pleuropneumoniae produce more than one toxin in vivo. To determine the value of the production of more than one toxin in the development of disease, we tested the pathogenicity of isogenic strains of A. pleuropneumoniae serotype 1 that are mutated in the toxin genes apxIA and/or apxIIA or in the transport genes apxIBD. Bacteria mutated in both apxIA and apxIIA, or in apxIBD, were unable to induce pathological lesions, thereby confirming the conclusion that ApxI and ApxII are essential for the pathogenesis of pleuropneumonia. Infection with isogenic strains lacking either ApxI or ApxII did not consistently lead to pleuropneumonia unlike the parent strain S4074. ApxII seemed at least as important as ApxI for the development of clinical and pathological symptoms. Only one of the four pigs inoculated with a mutant strain unable to produce ApxII developed mild pneumonia whereas two out of the three pigs inoculated with a mutant strain unable to produce ApxI developed more severe lesions. The results indicate that both ApxI and ApxII of A. pleuropneumoniae serotype 1 are necessary for full virulence. PMID- 15135510 TI - Serological relationship between cattle exposed to Brucella abortus, Yersinia enterocolitica O:9 and Escherichia coli O157:H7. AB - Sera from cattle naturally infected with Brucella abortus (n = 160), vaccinated with B. abortus S19 (n = 88) or immunized with Yersinia enterocolitica O:9 (n = 25) or Escherichia coli O157:H7 (n = 80) were collected. The sera were compared for antibody content to the same bacteria by indirect enzyme immunoassay (IELISA), fluorescence polarization assay (FPA) and competitive enzyme immunoassay (CELISA). Cattle sera (n = 523) collected randomly from across Canada were tested in the same tests. Sera from the B. abortus infected group reacted positively in the brucellosis IELISA (IELISA(Br)), CELISA and FPA (FPA(Br)) and the Y. enterocolitica IELISA (IELISA(Ye)) while the Y. enterocolitica FPA (FPA(Ye)) detected antibody in 93.8% and the E. coli IELISA (IELISA(Ec)) 86.9% and the E. coli FPA (FPA(Ec)) 48.1%. About 70% of the sera from B. abortus S19 vaccinated animals reacted in the three IELISAs, 45% in the CELISA, and 37.7% in the FPA(Ec), 21.6% in the FPA(Br) and 5.7% in the FPA(Ye). Sera from E. coli O:157 exposed cattle reacted mainly in the IELISA(Ec) and FPA(Ec) although surprisingly 87.5% reacted in the IELISA(Ye) and only 3.8% in the IELISA(Br). No reactions were observed with these sera in the FPA(Br) and FPA(Ye) but one serum gave a low positive reaction in the CELISA. All sera from Y. enterocolitica O:9 exposed cattle reacted in the IELISA(Br) and IELISA(Ye) and 80% in the IELISA(Ec). In the CELISA, 44% gave a positive reaction and 64% were positive in the FPA(Br), 28% in the FPA(Ye) and 12% in the FPA(Ec). Of the 523 Canadian sera, about 50% reacted in the E. coli tests with only minor reactions in the Y. enterocolitica O:9 and B. abortus assays. From the data, the cross reaction between E. coli O157:H7, Y. enterocilitica O:9 and B. abortus is dependent on the test used. Thus, extensive cross reaction was observed with the IELISA with much less reactivity in the FPA and the CELISA. PMID- 15135511 TI - Prevalence of Bartonella infection in wild African lions (Panthera leo) and cheetahs (Acinonyx jubatus). AB - Bartonella species are emerging pathogens that have been isolated worldwide from humans and other mammals. Our objective was to estimate the prevalence of Bartonella infection in free-ranging African lions (Panthera leo) and cheetahs (Acinonyx jubatus). Blood and/or serum samples were collected from a convenience sample of 113 lions and 74 cheetahs captured in Africa between 1982 and 2002. Whole blood samples available from 58 of the lions and 17 of the cheetahs were cultured for evidence of Bartonella spp., and whole blood from 54 of the 58 lions and 73 of the 74 cheetahs tested for the presence of Bartonella DNA by TaqMan PCR. Serum samples from the 113 lions and 74 cheetahs were tested for the presence of antibodies against Bartonella henselae using an immunofluorescence assay. Three (5.2%) of the 58 lions and one (5.9%) of the 17 cheetahs were bacteremic. Two lions were infected with B. henselae, based on PCR/RFLP of the citrate synthase gene. The third lion and the cheetah were infected with previously unidentified Bartonella strains. Twenty-three percent of the 73 cheetahs and 3.7% of the 54 lions tested by TaqMan PCR were positive for Bartonella spp. B. henselae antibody prevalence was 17% (19/113) for the lions and 31% (23/74) for the cheetahs. The prevalence of seropositivity, bacteremia, and positive TaqMan PCR was not significantly different between sexes and age categories (juvenile versus adult) for both lions and cheetahs. Domestic cats are thus no longer the only known carriers of Bartonella spp. in Africa. Translocation of B. henselae seronegative and TaqMan PCR negative wild felids might be effective in limiting the spread of Bartonella infection. PMID- 15135512 TI - Characterization of a 39kDa capsular protein of avian Pasteurella multocida using monoclonal antibodies. AB - The role of a 39kDa protein of avian Pasteurella multocida in pathogenesis of fowl cholera was investigated using monoclonal antibodies (Mabs). Mabs were prepared by immunization of BALB/c mice with a crude capsular extract (CCE) of P. multocida strain P-1059 (serovar A:3). Totally eight hybridomas producing Mab were obtained. Immunoblot analysis of the hybridomas revealed that all the Mabs recognized a 39kDa protein of CCE. Treatment of CCE antigen with proteinase K or periodic acid indicated that the epitope recognized was proteinaceous. The Mabs reacted with a major 39kDa protein of CCE from encapsulated strains but not with any protein of non-capsulated strains indicating that a direct correlation between encapsulation and the 39kDa protein. Immunoelectron microscopy on strain P-1059 and the non-capsulated derivative P-1059B (serovar -:3) reacting with the Mabs and gold-labeled anti-mouse IgG indicated that the protein is associated with the capsule. The Mabs significantly inhibited the adherence of encapsulated P. multocida strains to chicken embryo fibroblast cells, but only slightly that of non-capsulated strains. Mice passively immunized with the Mabs were protected from lethal challenge with virulent strains P-1059 and X-73 (serovar A:1). Thus the capsular 39kDa protein was determined to be an adherence factor and a cross protective antigen of avian P. multocida type A strains. PMID- 15135513 TI - Molecular conservation of MSP4 and MSP5 in Anaplasma marginale and A. centrale vaccine strain. AB - Anaplasma centrale msp4 and msp5 genes were cloned and sequenced, and the recombinant proteins were expressed. The identity between Anaplasma marginale and A. centrale MSP4 was 83% in the nucleotide sequences and 91.7% in the encoded protein sequences. A. centrale msp5 nucleotide sequences shared 86.8% identity with A. marginale msp5, and there was 92.9% homology between A. centrale and A. marginale encoded amino acids of the MSP5 protein. Southern blots hybridized with probes derived from the msp4 and msp5 central regions indicate that msp4 and msp5 of A. centrale are encoded by single copy genes. Recombinant MSP4 and MSP5 fusion proteins reacted with anti-A. marginale monoclonal antibodies ANAR76A1 and ANAF16C, respectively, demonstrating the conservation of conformation-sensitive B cell epitopes between A. centrale and A. marginale. These data demonstrate the structural and antigenic conservation of MSP4 and MSP5 in A. centrale and A. marginale. This conservation is consistent with the cross-protective immunity between A. marginale and A. centrale and supports the development of improved vaccines based upon common outer membrane proteins. PMID- 15135515 TI - Development of a polymerase chain reaction for the detection of Mycoplasma felis in domestic cats. AB - Mycoplasma felis is associated with conjunctivitis and respiratory disease in domestic cats. Currently no rapid diagnostic test is available for the detection of M. felis in clinical samples that does not rely on prior cultivation of the organism. The objective of this study was to determine whether a polymerase chain reaction (PCR) based upon the 16S/23S rRNA intergenic spacer sequence is suitable for the identification of M. felis directly in feline clinical samples. The high conservation between the 16S/23S rRNA intergenic spacers (IGS) of differing isolates of M. felis was established by sequence analysis and a PCR was developed to this region by comparison to IGS of other mycoplasmas. The PCR was found to be highly specific for M. felis and further PCR analysis on clinical samples showed the PCR to be highly sensitive and more rapid than the other methods of identification currently available. PMID- 15135514 TI - Efficacy of different commercial and new inactivated vaccines against ovine enzootic abortion. AB - The protective efficacy of two inactivated commercial (A, B) and two new inactivated vaccines (M7, QS) against ovine enzootic abortion was determined in two separate experiments in sheep. Vaccine A contained chlamydiae propagated in chicken embryos, adjuvated with Marcol 82, and vaccine B contained chlamydiae cultured in cell monolayers, adjuvated with aluminium hydroxide. For the preparation of the experimental vaccines, Chlamydophila abortus AB7 strain was cultured in McCoy cells and adjuvated with QS-21 (QS) or Montanide ISA 773 (M7). The ewes were vaccinated twice subcutaneously and challenged at 90 days of gestation. Protection was evaluated by clinical, bacteriological and serological examinations, and compared to two control groups: one of infected but not vaccinated ewes, and another of vaccinated but not infected ewes. The experimental vaccines induced considerably better protection than the two commercial ones. The new vaccine M7 especially showed no abortions, a good antibody response, the highest newborn lamb weights and the lowest level of C. abortus shedding at lambing. PMID- 15135516 TI - Susceptibility of different bacterial species isolated from food animals to copper sulphate, zinc chloride and antimicrobial substances used for disinfection. AB - A total of 569 different bacterial isolates (156 Salmonella, 202 E. coli, 43 S. aureus, 38 S. hyicus, 52 E. faecalis, 78 E. faecium) were tested for susceptibility to copper sulphate, benzalkonium chloride, hydrogen peroxide and chlorhexidine using MIC determinations. A total of 442 isolates were also tested for susceptibility to formaldehyde and 177 isolates for susceptibility to zinc chloride. Enterococcal isolates formed a bimodal distribution of MICs to copper sulphate, whereas the other bacterial species formed one large population. Otherwise the isolates formed one large population of susceptibilities to the different antimicrobial agents. Large variations were observed in the susceptibility of the different bacterial species to the different compounds. Staphylococci were in general very susceptible to all antimicrobial compounds tested. The Salmonella isolates were in general less susceptible to copper sulphate, benzalkonium chloride and chlorhexidine followed by E. coli and the Gram-positive species. The opposite was the case for zinc chloride. All isolates were very susceptible to H(2)O(2) with MICs ranging from 0.002 to 0.016%, and to formaldehyde with MICs at 0.003 and 0.006%. This study showed that Danish bacterial isolates from livestock so far have not or have only to a limited degree developed resistance to antimicrobial compounds commonly used for disinfection. Acquired copper resistance was only found in enterococci. There were large differences in the intrinsic susceptibility of the different bacterial species to these compounds, and Salmonella especially seems intrinsically less susceptible than the other bacterial species, which might have human health implications. PMID- 15135517 TI - Evaluation of three serum i-ELISAs using monoclonal antibodies and protein G as peroxidase conjugate for the diagnosis of bovine brucellosis. AB - Three i-ELISAs using LPS, the immunodominant component of Brucella abortus, were developed with three different conjugates: monoclonal antibodies 1C8 (anti-bovine IgG(1)) and 3H3 (mainly specific for bovine IgG(2) but also reacting with IgG(1)) and protein G (reacts with both bovine IgG subclasses). Using a cut-off value of 2.5U/ml, the i-ELISA with 3H3 as conjugate had a specificity (95% CI: 98.32 99.63%) that was significantly higher than the same assay with 1C8 (95% CI: 96.08 98.26%) or PG (95% CI: 95.83-98.09%). In areas where false positive serological reactions (FPSR) were common, the specificity of the i-ELISAs decreased significantly. The specificity of the i-ELISAs increased with the age of the animals tested, irrespective of the conjugate. The specificity of the i-ELISAs and traditional tests was also examined using sera from animals infected per os with bacteria bearing LPS similar to the Brucella LPS. It appeared that Yersinia enterocolitica O:9, Xanthomonas maltophilia and Salmonella urbana infections induced FPSR both in the i-ELISAs and in the traditional tests, but the 3H3 assay was significantly less prone to produce false positive reactions than the 1C8 and PG assays. The i-ELISAs were more sensitive, allowed earlier detection, and were more persistent than the traditional serological tests both in experimentally and naturally Brucella-infected animals. Weekly i-ELISA monitoring of experimentally infected pregnant heifers (previously vaccinated or not) allowed a prediction of abortion. Furthermore, the 1C8 assay showed significantly higher titres irrespective of day post-infection and vaccination status. The accuracy of the assay could be improved by making use of additional information (e.g. animal age or conjugate) and by selecting appropriate cut-off points on the basis of the prevailing epidemiological situation. The i-ELISAs appear an appropriate choice in order to maintain an official brucellosis-free status because of their sensitivity, early detection and long persistence and, for the same reasons, seem especially valuable for the detection of latent carriers (i.e. animals classified negative by classical serological tests) among imported animals. PMID- 15135518 TI - Selection of enterococci for potential canine probiotic additives. AB - Enterococci are important inhabitants of animal intestine and are widely used in probiotic products. A potentially successful probiotic strain is expected to have several desirable properties in order to be able to exert its beneficial effects. Forty enterococcal isolates from dog faeces were tested for characters believed to be important for probiotic strains; bacteriocin production, resistance or tolerance to antibiotics, low pH, bile tolerance and adhesive activity. The total count of enterococci was found to be 3.3-7.3log(10)CFU/g of faeces. Most identified strains were Enterococcus faecium. All strains were sensitive to vancomycin, ampicillin, penicillin and chloramphenicol. Thirty-three percentage of strains were resistant to erythromycin and 28% to tetracycline. Among 40 isolates, 75% showed a broad inhibitory spectrum only against Gram-positive indicator bacteria. Seven strains with broad bacteriocin activity were selected for further assays. In the presence of 1% bile, the survival rate of selected strains ranged between 72 and 98%. Survival of strains at pH 3.0 was found in the range between 76 and 87% after 3h. The adhesion of the tested strains to intestinal mucus ranged from 4 to 11% for canine mucus and from 5 to 8% for human mucus. E. faecalis EE4 and E. faecium EF01 showed the best probiotic properties. It indicates that they could be used as new candidate probiotic strains after in vivo testing. PMID- 15135519 TI - Higher incidence of Malassezia pachydermatis in the eyes of dogs with corneal ulcer than in healthy dogs. AB - Malassezia pachydermatis is usually associated with otitis and dermatitis in dogs but it can also cause diseases in other species, including humans. In a human neonatal intensive care unit, M. pachydermatis was isolated from an infant's ocular discharge. Therefore, the aim of this study was to ascertain the presence of Malassezia spp. and its possible consequences in dogs' eyes. This research included 19 dogs with unilateral or bilateral corneal ulcers and 60 healthy dogs. A total of 158 clinical specimens from both the groups were obtained from the conjunctival sac of each eye by a calibrated platinum loop. The samples were placed on Dixon and blood agar, incubated at 35 degrees C, and examined daily for 15 days. Then, the strains were subcultured on Sabouraud agar. Of 22 clinical specimens collected from the eyes with corneal ulcers, five cultures (23%) were positive for M. pachydermatis. Of 16 samples collected from the contralateral healthy eye, cultures were positive in three samples (19%). Three animals had unilateral corneal ulcer and positive cultures for M. pachydermatis in both the eyes. Two dogs had unilateral corneal ulcer and positive cultures for M. pachydermatis in the same eye. However, from the 120 samples of 60 healthy dogs, only four clinical specimens (3%) had positive cultures for M. pachydermatis. The findings of M. pachydermatis, in a considerable percentage of clinical specimens from dogs with corneal ulcer, suggest its possible role at least as an aggravating factor in the pathophysiology of this disease. PMID- 15135520 TI - Identification and differentiation of avirulent and virulent Rhodococcus equi using selective media and colony blotting DNA hybridization to determine their concentrations in the environment. AB - Selective agar media have been used for many years to facilitate the isolation of Rhodococcus equi from environmental and clinical samples. However, characterisation of R. equi still requires the use of immunochemical or polymerase chain reaction (PCR) analysis to differentiate between virulent and avirulent isolates. Here, we describe a novel method to detect and differentiate between R. equi isolates using colony blotting and DNA hybridization. Radiolabelled PCR product derived from the R. equi rrnA gene and specific hybridization conditions enabled differentiation of colonies of R. equi from environmental species, whilst radiolabelled PCR product derived from the R. equi vapA gene, under specific hybridization conditions, allowed differentiation between avirulent and virulent R. equi. This technique has the potential to be used for quantitative screening of large environmental and clinical samples for both avirulent and virulent R. equi. Its use in ecological and epidemiological studies of R. equi has the potential to improve understanding of the relationship between the environment, the foal and the disease. PMID- 15135521 TI - Comparison of an interferon-gamma to a phospholipase D enzyme-linked immunosorbent assay for diagnosis of Corynebacterium pseudotuberculosis infection in experimentally infected goats. AB - The optimal method of control of caseous lymphadenitis of goats caused by Corynebacterium pseudotuberculosis is eradication of infection by identification and removal of infected carrier animals. The objective of this study was to compare detection of C. pseudotuberculosis experimentally infected goats using a commercially available bovine interferon-gamma (IFN-gamma) whole blood enzyme linked immunosorbent assay (ELISA) to serological response to a recombinant phospholipase D (PLD) ELISA. The tests were assessed repeatedly over 1 year in three infected and three non-infected goats. Using a IFN-gamma optical density cut-off at 0.10 as positive under the conditions used, the test accurately detected C. pseudotuberculosis experimentally infected goats over a 363 day period with a reliability of 89.2% and non-infected goats with a reliability of 97.1%. Using a cut-off value of the mean for negative samples plus two standard deviations, the PLD ELISA detected C. pseudotuberculosis experimentally infected goats over this period with a reliability of 81.0% and non-infected goats with a reliability of 97.0%. The PLD ELISA was however more predictive than the IFN gamma ELISA of the presence of lesions observed at postmortem examination of infected goats. PMID- 15135522 TI - Targeting internal ribosome entry site (IRES)-mediated translation to block hepatitis C and other RNA viruses. AB - A number of RNA-containing viruses such as hepatitis C (HCV) and poliovirus (PV) that infect human beings and cause serious diseases use a common mechanism for synthesis of viral proteins, termed internal ribosome entry site (IRES)-mediated translation. This mode of translation initiation involves entry of 40S ribosome internally to the 5' untranslated region (UTR) of viral RNA. Cap-dependent translation of cellular mRNAs, on the other hand, requires recognition of mRNA 5' cap by the translation machinery. In this review, we discuss two inhibitors that specifically inhibit viral IRES-mediated translation without interfering with cellular cap-dependent translation. We present evidence, which suggest that one of these inhibitors, a small RNA (called IRNA) originally isolated from the yeast Saccharomyces cerevisiae, inhibits viral IRES-mediated translation by sequestering both noncanonical transacting factors and canonical initiation factors required for IRES-mediated translation. The other inhibitor, a small peptide from the lupus autoantigen La (called LAP), appears to block binding of cellular transacting factors to viral IRES elements. These results suggest that it might be possible to target viral IRES-mediated translation for future development of therapeutic agents effective against a number of RNA viruses including HCV that exclusively use cap-independent translation for synthesis of viral proteins. PMID- 15135523 TI - Diversity of genetic environment of bla(CTX-M) genes. AB - Twenty-four non-clonally related enterobacterial isolates producing the emerging CTX-M-type extended-spectrum beta-lactamases were recovered from several countries including France, India, Poland, and Turkey. They had been isolated from 2000 to 2003. beta-Lactamases CTX-M-2, CTX-M-3, CTX-M-10, CTX-M-14 and CTX-M 15 were identified. Most of the isolates produced beta-lactamase CTX-M-15. Insertion sequence ISEcp1 was found upstream of bla(CTX-M-3), bla(CTX-M-10), bla(CTX-M-14) and bla(CTX-M-15) genes. A sequence similar to the inverted right repeat of ISEcp1 was identified downstream of bla(CTX-M-3), bla(CTX-M-10) and bla(CTX-M-15) genes suggesting the mobilization of these beta-lactamase genes by transposition events. In addition, Orf513 was identified upstream of the bla(CTX M-2) gene. This work further underlined widespread of bla(CTX-M-15) gene associated with ISEcp1. PMID- 15135524 TI - Novel GES/IBC extended-spectrum beta-lactamase variants with carbapenemase activity in clinical enterobacteria. AB - Two clinical isolates, an Escherichia coli and a Klebsiella pneumoniae, with decreased susceptibility to carbapenems were studied. This phenotype was associated with production of novel GES/IBC variant beta-lactamases, designated GES-3 (from E. coli) and GES-4 (from K. pneumoniae), exhibiting carbapenemase activity. Both enzymes possessed Ser at Ambler's position 170 instead of Gly found in the beta-lactamases GES-1 and IBC-1 that lack carbapenemase activity. Additionally, position 104 in GES-4 was occupied by a Lys as in IBC-1. bla(GES-3) and bla(GES-4) occurred as gene cassettes in the variable regions of class 1 integrons carried by plasmids. The structure of the GES-4-encoding integron was similar to that of previously described IBC-1 integrons. The GES-3-encoding integron was, most likely, truncated at the 3' conserved segment. PMID- 15135525 TI - Two FHA domains on an ABC transporter, Rv1747, mediate its phosphorylation by PknF, a Ser/Thr protein kinase from Mycobacterium tuberculosis. AB - Bacterial genomics have revealed the widespread occurrence of eukaryotic-like protein kinases in prokaryotes, but little is known about their regulation, endogenous substrates, and physiological role. The present study concerns one of these enzymes, the serine/threonine protein kinase PknF from Mycobacterium tuberculosis. It is shown that, in addition to its autokinase activity, PknF is able to phosphorylate Rv1747, a newly described ABC transporter. This reaction appears to involve two FHA domains of Rv1747. It is suggested that recruitment and phosphorylation of Rv1747 depend on the interaction between its two non redundant FHA domains and the autophosphorylated form of PknF. PMID- 15135526 TI - The nitrogen-fixing symbiotic bacterium Mesorhizobium loti has and expresses the gene encoding pyridoxine 4-oxidase involved in the degradation of vitamin B6. AB - The gene product of mll6785 of a nitrogen-fixing symbiotic bacterium Mesorhizobium loti MAFF303099 was identified as pyridoxine 4-oxidase, the first enzyme in the vitamin B6-degradation pathway. The gene was cloned and ligated into pET-21a+. Escherichia coli BL21(DE3) was co-transformed with the constructed plasmid plus pKY206 containing groESL genes encoding chaperonins. The overexpressed protein was purified to homogeneity by the ammonium sulfate fractionation and three chromatography steps. The enzymatic properties of the purified protein, such as K(m) values for pyridoxine (213+/-19 microM) and oxygen (78+/-10 microM), were compared to those of pyridoxine 4-oxidase from two bacteria with known vitamin B6-degradation pathway. M. loti grown in a Rhizobium medium showed the enzyme activity. The results suggest that M. loti also contains the degradation pathway of vitamin B6. PMID- 15135527 TI - Polyhydroxyalkanoate synthases PhaC1 and PhaC2 from Pseudomonas stutzeri 1317 had different substrate specificities. AB - The whole polyhydroxyalkanoate (PHA) synthesis gene locus of Pseudomonas stutzeri strain 1317 containing PHA synthase genes phaC1Ps, phaC2Ps and PHA depolymerase gene phaZPs was cloned using a PCR cloning strategy. The sequence analysis results of the phaC1Ps, phaC2Ps and phaZPs showed high homology to the corresponding pha loci of the known Pseudomonas strains, respectively. PhaC1Ps and PhaC2Ps were functionally expressed in recombinant Escherichia coli strains and their substrate specificity was compared. The results demonstrated that PhaC1Ps and PhaC2Ps from P. stutzeri 1317 had different substrate specificities when expressed in E. coli. In details, PhaC2Ps could incorporate both short-chain length 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoates (mcl 3HA) into PHA, while PhaC1Ps only favored mcl 3HA for polymerization. PMID- 15135528 TI - Cloning and characterization of the gene encoding the z66 antigen of Salmonella enterica serovar Typhi. AB - Z66 antigen-positive strains of Salmonella enterica serovar Typhi change flagellin expression in only one direction from the z66 antigen to the d or j antigen, which is different from the phase variation of S. enterica serovar Typhimurium. In the present study, we identified a new flagellin gene in z66 antigen-positive strains of S. enterica serovar Typhi. The genomic structure of the region containing this new flagellin gene was similar to that of fljBA operon of biphasic S. enterica serovars. A fljA-like gene was present downstream of the new flagellin gene. A rho-independent terminator was located between the new flagellin gene and the fljA-like gene. Hin-like gene was not present upstream of the new flagellin gene. We generated a mutant strain of S. enterica serovar Typhi, which carries a deletion of the new flagellin gene. Western blotting revealed that the 51-kDa z66 antigen protein was absent from the population of proteins secreted by the mutant strain. Southern hybridization demonstrated that the z66 antigen-positive strains of S. enterica serovar Typhi carried the new flagellin gene and fliC on two different genomic EcoRI fragments. When z66 antigen-positive strains were incubated with anti-z66 antiserum, the flagellin expression by S. enterica serovar Typhi changed from z66 antigen to j antigen. The new flagellin gene and the fljA-like gene were absent in the strain with altered flagellin expression. These results suggested that the new flagellin gene is a fljB-like gene, which encodes the z66 antigen of S. enterica serovar Typhi, and that deletion of fljBA-like operon may explain why S. enterica serovar Typhi alters the flagellin expression in only one direction from the z66 antigen to the d or j antigen. PMID- 15135529 TI - Purification and characterization of aminopeptidase (pumAPE) from Ustilago maydis. AB - The aminopeptidase pumAPE was purified from the haploid fungus Ustilago maydis FB1 strain. The purification procedure consisted of ammonium sulfate fractionation and three chromatographic steps, which included anion-exchange, hydrophobic interaction, and gel filtration chromatography, resulting in a 23% recovery. The molecular mass of the dimeric enzyme was estimated to be 110 kDa and 58 kDa by gel filtration chromatography and SDS-PAGE, respectively. Enzymatic activity was optimal at pH 7.0 and at 35 degrees C toward Lys-pNA and the pI was determined to be 5.1. The enzyme was inhibited by EDTA-Na2, 1,10- phenanthroline, bestantin, PMSF and several divalent cations (Cu2+, Hg2+ and Zn2+). The aminopeptidase showed a preference for lysine and arginine in the N-position. The K(m) value was 54.4 microM and the Vmax value was 408 micromolmin(-1)mg(-1) for Lys-pNA. PMID- 15135530 TI - Fungal cytochrome P450s catalyzing hydroxylation of substituted toluenes to form their hydroxymethyl derivatives. AB - The degradation of a series of nitroaromatic compounds by the lignin-degrading fungus Phanerochaete chrysosporium was examined. From 4-nitrotoluene (4-NT), several metabolic intermediates were identified. Initially, 4-NT was converted to 4-nitrobenzyl alcohol (4-NBA), followed by the oxidation reactions to form 4 nitrobenzaldehyde and 4-nitrobenzoic acid, albeit slowly. Exogenously added 4 nitrobenzaldehyde and 4-nitrobenzoic acid were predominantly reduced to 4-NBA. The fungal formation of 4-NBA was inhibited by piperonyl butoxide, a cytochrome P450 inhibitor, suggesting the involvement of cytochrome P450 in the hydroxylation of the methyl group. Similarly, 2-, and 3-nitrotoluenes and 4 chlorotoluene were converted to the corresponding arylalcohols by P. chrysosporium. On the other hand, toluene and 4-methoxytoluene were not converted. Thus, P. chrysosporium possesses an alkyl hydroxylation activity against aromatic compounds substituted with a strong electron-withdrawing group. PMID- 15135531 TI - Characterization of Cu- and Zn-containing superoxide dismutase of Rhodobacter sphaeroides. AB - Rhodobacter sphaeroides has a cuprozinc-containing superoxide dismutase (CuZnSOD) in its periplasm in addition to the cytoplasmic SOD that appears to contain iron (FeSOD). The FeSOD is constitutively expressed under the growth conditions examined, whereas the CuZnSOD is detected only when the light-harvesting complexes are found. The CuZnSOD expression is regulated post-transcriptionally, and the enzyme appears to protect the photoheterotrophic cells from periplasmic superoxide that may be generated by exposure of the cells to low O2 under the illuminated conditions at 10 W/m2. PMID- 15135532 TI - A new species of Pythium isolated from burgundian vineyards and its antagonism towards Botrytis cinerea, the causative agent of the grey mould disease. AB - During the course of an investigation on the pythiaceous fungi occurring in burgundian vineyards, three isolates of Pythium were found, all of which were characterised by spherical to lemoniform, proliferating sporangia. Only one out of these three isolates produced sexual characters (S-12) after prolonged incubation, but their asexual structures, temperature growth relationship and molecular characteristics were identical. Saprolegnia-type internal proliferation of the sporangia, and Phytophthora-type lemoniform, papillate sporangia were characteristic of all the three isolates. The type specimen is S-12 which is a medium slow growing saprophyte. The sequence of its ITS region of the rDNA shows a close relationship with the genus Phytophthora. Morphological and molecular characteristics of this isolate justifies its description as a new species: Pythium citrinum. When grown together with Botrytis cinerea, the causal agent of the grey mould disease of the grapevine, this new species shows a pronounced antagonism and suppresses its growth. This new species is very close to P. montanum, a recently described fungus and constitutes a "border species" linking the genera Pythium and Phytophthora. Morphological features of this new species, its antagonism with B. cinerea, the sequences of the ITS region of its nuclear ribosomal DNA, and its comparison with related species are discussed. PMID- 15135533 TI - Identification and genetic characterisation of orthopaedic Staphylococcus isolates collected in Italy by automated EcoRI ribotyping. AB - The aim of this study was to evaluate the possibility to use automated EcoRI ribotyping to address, during the same analysis, both identification and genetic characterisation of 38 Staphylococcus aureus and 64 coagulase-negative staphylococci collected from surgical injuries. The ribotyping identification results confirmed those obtained using the API Staph system for 96% of the isolates. All strains were successfully genotyped and the ribotyping discriminatory power, calculated using the Simpson's index of discrimination, was very high for both groups of staphylococci tested. The same, as well as different biotypes, were identified among isolates with the identical ribotyping profile. PMID- 15135534 TI - Age influences resistance of Caenorhabditis elegans to killing by pathogenic bacteria. AB - Caenorhabditis elegans has previously been proposed as an alternative host for models of infectious disease caused by human pathogens. When exposed to some human pathogenic bacteria, the life span of nematodes is significantly reduced. We have shown that mutations in the age-1, and/or age-2 genes of C. elegans, that normally enhance life expectancy, can also increase resistance to killing by the bacterial pathogens Pseudomonas aeruginosa, Salmonella enterica var. Typhimurium, Burkholderia cepacia or Yersinia pseudotuberculosis. We also found that the rate at which wild-type C. elegans was killed by the bacterial pathogens tested increased as nematodes aged. In the case of P. aeruginosa infection, the difference in life span of wild type and age-1 mutants of C. elegans was not due to differences in the level of bacterial colonisation of the gut. PMID- 15135535 TI - Brucella abortus nicotinamidase (PncA) contributes to its intracellular replication and infectivity in mice. AB - Brucella spp. are facultative intracellular pathogens that have the ability to survive and multiply in professional and non-professional phagocytes, and cause abortion in domestic animals and undulant fever in humans. The mechanism and factors of virulence are not fully understood. Nicotinamidase/pyrazinamidase mutant (pncA mutant) of Brucella abortus failed to replicate in HeLa cells, and showed a lower rate of intracellular replication than that of wild-type strain in macrophages. Addition of nicotinic acid, but not nicotinamide, into medium supported intracellular replication of pncA mutant in HeLa cells and macrophages. The pncA mutant was not co-localizing with either late endosomes or lysosomes. The B. abortus virB4 mutant was completely cleared from the spleens of mice after 4 weeks, while the pncA mutant showed a 1.5-log reduction of the number of bacteria isolated from spleens after 10 weeks. Although pncA mutant showed reduced virulence in mice and defective intracellular replication, its ability to confer protection against the virulent B. abortus strain 544 was fully retained. These results suggest that PncA does not contribute to intracellular trafficking of B. abortus, but contributes to utilization of nutrients required for intracellular growth. Our results indicate that detailed characterizations of the pncA mutant may help the improvement of currently available live vaccines. PMID- 15135536 TI - Use of a novel allele of the Escherichia coli aacC4 aminoglycoside resistance gene as a genetic marker in mycobacteria. AB - The aacC4 gene from Escherichia coli can be expressed in mycobacteria and confers resistance to apramycin. However, the major limitation of the aacC4 gene as a genetic tool is that the gene also confers resistance to kanamycin and gentamicin, two antibiotics commonly used for selection in mycobacterial genetics, thus reducing the utility of the aacC4 gene in the mycobacterial field. To overcome this problem we constructed, by chemical mutagenesis, a mutant allele of the E. coli aacC4 gene that still confers resistance to apramycin but has a reduced ability to confer resistance to kanamycin and gentamicin. We then constructed a variety of E. coli-mycobacteria shuttle plasmids containing this mutant allele. PMID- 15135537 TI - Topological studies on the twin-arginine translocase component TatC. AB - The twin-arginine translocation (Tat) system can translocate folded proteins across biological membranes. Among the known Tat-system components in Escherichia coli, TatC is the only protein with multiple trans-membrane domains. TatC is important for translocon interactions with Tat substrates. The knowledge of its membrane topology is therefore crucial for the understanding of substrate binding and translocon function. Recently, based on active PhoA reporter fusions to the second predicted cytoplasmic loop of TatC, a topology with four trans-membrane domains has been suggested, calling in silico predictions of six trans-membrane domains into question. Here we report studies with translational fusions of TatC to the topological marker enzymes PhoA and LacZ which provide strong evidence for a six-trans-membrane domain topology. The stop transfer capacity of the fourth trans-membrane domain was found to be strongly influenced by the succeeding cytoplasmic domain. The presence of linker sequences at PhoA-fusion sites of the cytoplasmic domain induced PhoA leakage. In the case of one tested fusion (S185 PhoA), the stop-transfer efficiency was already low due to the negative charge in the center of the fourth trans-membrane domain (E170). The results point to the importance of cytoplasmic loops for the stabilization of stop-transfer sequences and revoke evidence for only four trans-membrane domains of TatC. PMID- 15135538 TI - The von Willebrand factor-binding protein (vWbp) of Staphylococcus aureus is a coagulase. AB - Staphylococcus aureus encodes a secreted von Willebrand factor-binding protein (vWbp) of 482 amino acids. The N-terminal part of this protein is homologous to staphylocoagulase and therefore we investigated whether vWbp has coagulating activity. Recombinant vWbp was shown to coagulate human and porcine plasma efficiently, but was less active against plasma from other species. The coagulation efficiency was concentration dependent, and could be inhibited by specific antibodies against vWbp. Furthermore, the species-specific coagulation by vWbp depended on the interaction with prothrombin. This interaction also resulted in specific cleavage of vWbp, releasing the C-terminal part from the coagulating domain. PMID- 15135539 TI - An ffh mutant of Streptococcus mutans is viable and able to physiologically adapt to low pH in continuous culture. AB - Previously, we described in Streptococcus mutans strain NG8 a 5-gene operon (sat) that includes ffh, the bacterial homologue of the eukaryotic signal recognition particle (SRP) protein, SR54. A mutation in ffh resulted in acid sensitivity but not loss of viability. In the present study, chemostat-grown cells of the ffh mutant were shown to possess only 26% and 39% of the parental membrane F-ATPase activity and 55% and 75% of parental glucose-phosphotransferase (PTS) activity when pH-7 and pH-5-grown cells, respectively, were assayed. Two-dimensional-gel electrophoretic analyses revealed significant differences in protein profiles between parent and ffh-mutant strains at both pH 5 and pH 7. It appears that the loss of active SRP (Ffh) function, while not lethal, results in substantial alterations in cellular physiology that includes acid tolerance. PMID- 15135540 TI - An improved method for deleting large regions of Escherichia coli K-12 chromosome using a combination of Cre/loxP and lambda Red. AB - We have established an improved large deletion method in Escherichia coli genome using a combination of two different recombination systems, lambda Red and Cre/loxP. The loxP site could be rapidly and efficiently integrated in the genome by lambda Red and large deletions of both 117- and 165-kbp regions could be generated in 100% efficiency by Cre/loxP. Comparative genomic hybridization microarray experiments of deletion strains indicated that deletions were generated only in expected regions of the genome. These results have demonstrated that the method is useful for genome engineering in E. coli. PMID- 15135541 TI - NF-kappaB activation suppresses host cell apoptosis during Rickettsia rickettsii infection via regulatory effects on intracellular localization or levels of apoptogenic and anti-apoptotic proteins. AB - Rickettsia rickettsii, a gram-negative and obligate intracellular bacterium, is the causative agent of Rocky Mountain spotted fever. In human infections, the primary target of R. rickettsii infection is vascular endothelium. Our laboratory has shown that activation of nuclear transcription factor-kappa B (NF-kappaB) during R. rickettsii infection of cultured human endothelial cells protects against apoptosis by preventing the activation of apical caspases-8 and -9, and the effector caspase-3. To understand upstream signaling mechanisms, we have determined the effect of NF-kappaB blockade on the status of different Bcl-2 (B cell lymphoma 2) proteins in this study. Quantitative analysis following TUNEL and Hoechst staining confirmed that infection of endothelial cells with R. rickettsii for 6 h in the presence of a specific NF-kappaB inhibitor, MG132, resulted in induction of apoptosis. Infection-induced apoptosis of EC was associated with decreased level of Bid and accumulation of Bad, while cytosolic level of Bax remained relatively unchanged. Further, the cellular levels of apoptosis antagonist Bcl-2 were found to be down-regulated and apoptogenic mitochondrial proteins Smac and cytochrome c were released into cytoplasm. These results implicate an important regulatory role for NF-kappaB in controlling the intracellular levels and/or localization of pro- as well as anti-apoptotic proteins of Bcl-2 family, the intricate balance of which is a critical determinant of downstream signaling mechanisms governing cell fate during intracellular infection. PMID- 15135542 TI - The effect of NAG-thiazoline on morphology and surface hydrophobicity of Escherichia coli. AB - The beta-hexosaminidase inhibitor and structural analog of the putative oxazolium reaction intermediate of lytic transglycosylases, N-acetylglucosamine thiazoline (NAG-thiazoline), was synthesized in 46% overall yield and tested as an inhibitor of Escherichia coli growth. NAG-thiazoline, at concentrations up to 1 mg/ml, was not found to affect the viability of E. coli DH5alpha. However, the compound did induce morphological changes to the cells. Growth of cells in the presence of NAG thiazoline caused an apparent inhibition of the biosynthesis of the cylindrical regions of the cells such that they became much shorter in length. The surface of these shorter cells was found to be much less hydrophobic compared to untreated cells as determined by the bacterial adhesion to hydrocarbon (BATH) assay. In addition, the co-administration of NAG-thiazoline with 1.7 x MIC concentrations of ampicillin prevented cell lysis suggesting that the compound inhibited autolytic enzymes, in particular the lytic transglycosylases. PMID- 15135543 TI - Isolation of Elaeagnus-compatible Frankia from soils collected in Tunisia. AB - The occurrence and diversity of Frankia nodulating Elaeagnus angustifolia in Tunisia were evaluated in 30 soils from different regions by a Frankia-capturing assay. Despite the absence of actinorhizal plants in 24 of the 30 soils, nodules were captured from all the samples. Eight pure strains were isolated from single colonies grown in agar medium. On the basis of 16S rRNA and GlnII sequences, seven strains were clustered with Frankia, colonizing Elaeagnaceae and Rhamnaceae in two different phylogenetic groups while one strain described a new lineage in the Frankia assemblage, indicating that Frankia strains genetically diverse from previously known Elaeagnus-infective strains are present in tunisian soils. Genomic fingerprinting determined by rep-PCR, and tDNA-PCR-SSCP, confirmed the wide genetic diversity of the strains. PMID- 15135544 TI - Attenuation regulation of amino acid biosynthetic operons in proteobacteria: comparative genomics analysis. AB - Candidate attenuators were identified that regulate operons responsible for biosynthesis of branched amino acids, histidine, threonine, tryptophan, and phenylalanine in gamma- and alpha-proteobacteria, and in some cases in low-GC Gram-positive bacteria, Thermotogales and Bacteroidetes/Chlorobi. This allowed us not only to describe the evolutionary dynamics of regulation by attenuation of transcription, but also to annotate a number of hypothetical genes. In particular, orthologs of ygeA of Escherichia coli were assigned the branched chain amino acid racemase function. Three new families of histidine transporters were predicted, orthologs of yuiF and yvsH of Bacillus subtilis, and lysQ of Lactococcus lactis. In Pasteurellales, the single bifunctional aspartate kinase/homoserine dehydrogenase gene thrA was predicted to be regulated not only by threonine and isoleucine, as in E. coli, but also by methionine. In alpha proteobacteria, the single acetolactate synthase operon ilvIH was predicted to be regulated by branched amino acids-dependent attenuators. Histidine biosynthetic operons his were predicted to be regulated by histidine-dependent attenuators in Bacillus cereus and Clostridium difficile, and by histidine T-boxes in L. lactis and Streptococcus mutans. PMID- 15135545 TI - Swm1p, a subunit of the APC/cyclosome, is required to maintain cell wall integrity during growth at high temperature in Saccharomyces cerevisiae. AB - Swm1p, a subunit of the APC cyclosome, was originally identified for its role in the later stages of the sporulation process and is required for spore wall assembly. In addition, this protein is required to maintain cell wall integrity in vegetative cells during growth at high temperature. Electron microscopy analyses of mutant cells grown at the restrictive temperature in the absence of osmotic support show that the cell wall is clearly abnormal, with large number of discontinuities that may be responsible for the observed lysis. The mutant cells show a 7-fold reduction in glucan synthase activity during growth at 38 degrees C and a 3.5-fold increase in the chitin content of the cell wall. The chitin is deposited in a delocalized manner all over the cell wall, where it accumulates in patches in abnormal regions. The excess chitin is mainly synthesized by the action of chitin synthase III (Chs3p), since it disappears in the swm1 chs3 double-mutant. PMID- 15135546 TI - Stress-dependent regulation of the gene encoding thioredoxin reductase from the fission yeast. AB - The unique putative gene for thioredoxin reductase (TrxR) was isolated from the chromosomal DNA of the fission yeast Schizosaccharomyces pombe. The determined DNA sequence carries 3125 bp, and encodes the plausible 322 amino acid sequence of TrxR with a molecular mass of 34,618 Da. The S. pombe cells harboring the cloned TrxR gene contain increased TrxR activity, and shows higher survivals on solid media with mercuric chloride or aluminum chloride. The 1526 bp upstream region was fused into promoterless beta-galactosidase gene of the shuttle vector YEp367R to generate the fusion plasmid. The synthesis of beta-galactosidase from the fusion plasmid pYUTR10 was enhanced by menadione, mercuric chloride, hydrogen peroxide, aluminium chloride and sodium selenite. Menadione significantly enhanced the TrxR mRNA level in the S. pombe cells, which was detected by RT-PCR. Induction of the S. pombe TrxR gene by menadione and mercuric chloride occurs through the mediation of the transcription factor Pap1. These results suggest that the S. pombe TrxR gene is one of the stress response-related genes. PMID- 15135547 TI - Understanding hard-core drug use among urban Puerto Rican women in high-risk neighborhoods. AB - The objectives of the current study were to determine the incidence and prevalence of hard-core drug (HCD) use and to explore the predictors of HCD use in a three-wave longitudinal study of women ages 18-35 living in urban areas of high drug traffic in Puerto Rico. Prevalence of HCD use was determined by positive self-report or positive toxicological tests for crack, cocaine, or heroine at baseline and follow-up periods. Incidence density was calculated using Poisson methods. Predictors of HCD use were identified using the generalized estimating equation (GEE) approach. Prevalence rates fluctuated between 16.3 at baseline and 12.6 and 14.6 at subsequent waves. Incidence of crack/cocaine or heroine ranged from 7.4 to 6.3 per 100 person years. Low education, unemployment, current alcohol use, and severe partner violence predict incident HCD use. Therefore, prevention programs for HCD use must consider strategies to promote social mobility and empowerment and to mitigate violence against women. PMID- 15135548 TI - Double-blind trial of the effects of tryptophan depletion on depression and cerebral blood flow in smokers. AB - Studies of clinically depressed patients have documented left frontal lobe hypoactivity. Smokers also show an increased prevalence of depression and evidence that nicotine normalizes qEEG indices of left frontal lobe activity. Tryptophan depletion (TD) has been shown to increase negative mood in smokers, particularly those with recurrent depression. Thus, in smokers, we expected that increased depression during TD would be associated with decreased cerebral blood flow, specifically in the left frontal lobe. Hamilton depression scores and relative regional cerebral blood flow (rCBF) were measured with SPECT using (99m)Tc-hexamethylpropyleneamineoxime in seven smokers after TD and after a control procedure. Decreased bilateral cerebral blood flow to the inferior frontal (IF) lobe following TD relative to placebo was associated with increased depressed mood (r= -.653, P<.05). Among smokers, a decrease in brain serotonin is associated with increased depressed mood and with focal bilateral decreases in IF activity. Chronic nicotine exposure appears to be associated with cortical responses suggestive of depressive vulnerability. PMID- 15135550 TI - Statistical definition of relapse: case of family drug court. AB - At any point in time, a patient's return to drug use can be seen either as a temporary event or as a return to persistent use. There is no formal standard for distinguishing persistent drug use from an occasional relapse. This lack of standardization persists although the consequences of either interpretation can be life altering. In a drug court or regulatory situation, for example, misinterpreting relapse as return to drug use could lead to incarceration, loss of child custody, or loss of employment. A clinician who mistakes a client's relapse for persistent drug use may fail to adjust treatment intensity to client's needs. An empirical and standardized method for distinguishing relapse from persistent drug use is needed. This paper provides a tool for clinicians and judges to distinguish relapse from persistent use based on statistical analyses of patterns of client's drug use. To accomplish this, a control chart is created for time-in-between relapses. This paper shows how a statistical limit can be calculated by examining either the client's history or other clients in the same program. If client's time-in-between relapse exceeds the statistical limit, then the client has returned to persistent use. Otherwise, the drug use is temporary. To illustrate the method, it is applied to data from three family drug courts. The approach allows the estimation of control limits based on the client's as well as the court's historical patterns. The approach also allows comparison of courts based on recovery rates. PMID- 15135549 TI - The blind spot in the nicotine replacement therapy literature: assessment of the double-blind in clinical trials. AB - While clinical trials of medications often use a double-blind procedure, the integrity of the blind and its relationship to treatment outcome is seldom examined. In this review, 73 double-blind, placebo-controlled clinical trials of the nicotine replacement therapies (NRTs) in smoking cessation were identified. Seventeen articles were found that assessed blindness integrity, demonstrating major variations in the assessment, analysis, and reporting of blindness integrity. Although 12 studies found that subjects accurately judged treatment assignment at a rate significantly above chance, the available literature does not permit definitive conclusions about blindness integrity. Recommendations for the assessment, analysis, and reporting of blindness integrity are made. PMID- 15135551 TI - History of marijuana use and tobacco smoking topography in tobacco-dependent adolescents. AB - Adolescent tobacco smokers have higher rates of marijuana (MJ) use than nonsmokers. Because MJ smoking typically involves deeper inhalation and longer breathholding than tobacco smoking, we hypothesized greater puff volume, longer puff duration and puff interval, and higher puff velocity during tobacco smoking among (1) MJ-using teens; (2) teens whose onset of MJ smoking occurred before tobacco (MBT). One hundred and three tobacco-dependent adolescents presented for smoking cessation treatment (66.0% female, 71.0% European American, mean age 15.3+/-1.25 years) smoked one cigarette of their own brand in the laboratory prior to study entry. Topography and associated physiological measures among current recreational (<5 days in a 14-day period) MJ users (n=25), current heavy (>/=5 days in a 14-day period) MJ users (n=22) and current non-MJ-smoking teens (n=56) were compared. There were no differences in tobacco smoking topography or physiological measures by recent MJ-smoking history or by order of substance initiation. Significantly more African American than European American adolescent smokers reported MJ use before tobacco. Our findings in adolescent smokers are consistent with results from adult studies in which history of MJ smoking was not associated with changes in tobacco smoking topography. PMID- 15135552 TI - Domestic violence in women with PTSD and substance abuse. AB - This study reports rates of domestic violence for women with posttraumatic stress disorder (PTSD) and substance dependence. The Conflict Tactics Scale-Revised (CTS2) was used to assess the rates of both physical and psychological aggression, lifetime and current, by self and partner. Results showed substantial rates, both lifetime and current by both partner and self and, for a substantial number in the sample, for multiple events within the past year. Consistent with prior studies, analyses indicated that the correlation between self and partner negative relationship behaviors was high. A significant association was found between past-month substance use and increased self and partner negative relationship behaviors. The complex dynamics of women as both recipients and enactors of negative relationship behaviors are discussed. PMID- 15135553 TI - Comparison of new alcohol use test, the Helsinki Alcohol Use Test questionnaire, and Alcohol Use Disorders Identification Test and laboratory markers serum gamma glutamyl transferase and carbohydrate-deficient transferrin. AB - The purpose of the present study was to compare and evaluate the new Helsinki Alcohol Use Test (HAUT) and Alcohol Use Disorders Identification Test (AUDIT) as screening instrument in the general working population. The relationship between the HAUT and serum gamma-glutamyl transferase (GGT), and carbohydrate-deficient transferrin (CDT) was also evaluated. Our results seem to indicate that the HAUT has good performance as screening instrument for alcohol-related problems in the general population. PMID- 15135554 TI - Time spent with friends who smoke and quit attempts among teen smokers. AB - Social interactions, as well as pharmacological effects, can reinforce smoking behavior in adolescents and pose challenges to smoking cessation. We hypothesized that time spent with friends who smoke would be inversely related to both the number and duration of previous quit attempts for teens receiving smoking cessation treatment. Demographic- and smoking-related data were collected from 98 adolescent smokers enrolled in an accruing smoking cessation study (68% female, 70% Caucasian, mean age 15.3 years). Among smokers who had at least one quit attempt, time spent with friends who smoke was inversely associated with the number of prior quit attempts (chi(2)=9.55, P=.0085) but not with their duration, suggesting a potential relationship between an adolescent's affiliation with smoking peers, smoking identity, and fewer quit attempts. PMID- 15135555 TI - Psychopathology and treatment outcome of drug dependent women in a perinatal program. AB - One hundred and five drug-dependent women in outpatient perinatal addiction treatment were classified by cluster analysis of the Minnesota Multiphasic Personality Inventory-2 (MMPI-2) profiles into high and low psychopathology (HP and LP) groups that differed on three validation measures. The HP group (n=29, 27.6%) had elevations on MMPI-2 Scales F, 2, 4, 6, 7, and 8, while the LP group (n=76, 72.4%) generated a normal range profile with elevations on Scales F and 4. Psychological outcomes differed by group. HP participants showed reduced alcohol, family, and psychiatric severity, and reduced depressive symptoms, while LP subjects showed reduced drug, self-debasing, and acting-out problems. Data suggest the need for lower intensity services for the majority of the perinatal drug dependent population with LP. PMID- 15135556 TI - Increased intensity of Ecstasy and polydrug usage in the more experienced recreational Ecstasy/MDMA users: a WWW study. AB - Recreational Ecstasy/MDMA (3,4-methylenedioxymethamphetamine) users often take a variety of psychoactive drugs, but there is little empirical data on how these drug consumption patterns change with greater experience of Ecstasy. The aim of this study was to compare the polydrug usage patterns reported by non-Ecstasy users, novice Ecstasy users, moderate Ecstasy users, and heavy Ecstasy users. In a WWW study of 763 unpaid volunteers, 481 had never taken Ecstasy, whereas 282 reported they had taken it. The Ecstasy users comprised 109 novice users (1-9 occasions), 136 moderate Ecstasy users (10-99 occasions), and 36 heavy Ecstasy users (+100 occasions). Each participant also reported their experience with a range of other psychoactive drugs. The Ecstasy users reported significantly greater psychoactive drug usage than the non-Ecstasy users. The novice, moderate, and heavy Ecstasy users also differed significantly from each other in the use of cocaine, amphetamine, LSD, and psilocybin mushrooms, but not of alcohol, cannabis, or cigarettes/nicotine. Experienced Ecstasy users also took significantly more MDMA tablets on each occasion, and reported a higher maximum weekly intake. The increased use of Ecstasy is associated with more intensive patterns of Ecstasy/MDMA intake, and the greater use of illicit CNS stimulants and hallucinogens, but not of alcohol, nicotine, or cannabis. These results are discussed in the context of cross-tolerance and drug predisposition/preference. PMID- 15135557 TI - High-risk relapse situations and self-efficacy: comparison between alcoholics and heroin addicts. AB - The main purpose of the present study is to investigate the differences between alcoholics and heroin addicts in some relapse dimensions and self-efficacy. The sample consisted of 180 inpatient males, who admitted to Al-Amal Hospital, Jeddah, Saudi Arabia, for treatment from substance abuse. The total sample was divided into two groups: 105 participants were heroin addicts and 75 were alcoholics. Two standardized questionnaires [inventory of drug taking situations (IDTS) and situational confidence questionnaire (SCQ)] were used. Heroin addicts have statistically significant high mean IDTS scores compared with the alcoholics sample, whereas the alcoholics have significantly higher mean SCQ scores than did their heroin counterpart. Again, the alcoholics had significantly lower admission and relapse rates compared with the heroin group. Furthermore, the results indicate that the highest high-risk relapse situations are negative emotions (NE), testing personal control (TPC), social pressure (SP), and urge and temptations (UT). PMID- 15135558 TI - Comparing stage of change measures in adolescent smokers. AB - This study compared stage of change measures among adolescent smokers. Participants were 56 adolescents who had received smoking tickets. They filled out an assessment packet including readiness to change measures [i.e., algorithm, Crittenden's measure, University of Rhode Island Change Assessment (URICA), and Ladder], smoking history, a 30-day calendar, Fagerstrom, self-efficacy measures, locus of control, and a problem screen. The Crittenden algorithm was correlated with the Ladder and URICA motivation scores as well as with decreased frequency/amount of smoking and past quit attempts. Ladder scores were correlated with less cigarettes smoked, self-efficacy, and fewer adolescent problems. Readiness to change was unrelated to nicotine dependence and locus of control. The classification of participants into stages by the Crittenden algorithm was associated with a significant MANOVA (Wilks' Lambda F=1.56, P<.025), with group differences on reported quit attempts, abstinence self-efficacy, and adolescent problems. The Crittenden measure and the URICA motivation score were sensitive to a MET intervention. PMID- 15135559 TI - The effects of controlled deep breathing on smoking withdrawal symptoms in dependent smokers. AB - This study was designed to assess the effect of controlled deep breathing on smoking withdrawal symptoms. In two laboratory sessions, dependent smokers refrained from smoking for 4 h. During a deep breathing session, participants were instructed to take a series of deep breaths every 30 min. During a control session, participants sat quietly. Controlled deep breathing significantly reduced smoking withdrawal symptoms, including craving for cigarettes and negative affect (tense, irritable), while resulting in the maintenance of baseline arousal (wide awake, able to concentrate) levels. Furthermore, a history of heavy smoking was associated with greater increases in arousal during the deep breathing session. The results of this preliminary study suggest that controlled deep breathing may be useful for relieving symptoms of smoking withdrawal. PMID- 15135560 TI - Personal strivings, binge drinking, and alcohol-related problems. AB - This study examined relations between personal strivings and alcohol use among college students. Personal strivings are ongoing goals that individuals are characteristically trying to achieve through their behavior. Participants generated lists of personal strivings following standard instructions and then completed an assessment of alcohol use and related problems. Participants returned to complete a follow-up assessment of drinking behavior after 30 days. Personal strivings were coded into content categories by trained raters using a coding manual. Four content categories were examined for this study: achievement, affiliation, health, and self-presentation. A series of t tests revealed that participants endorsing achievement strivings reported less alcohol-related problems and marginally fewer instances of binge drinking during the 30-day follow-up period. In contrast, participants endorsing self-presentation strivings reported more alcohol-related problems during the follow-up period. PMID- 15135561 TI - Motivational stages of adolescent smoking initiation: predictive validity and predictors of transitions. AB - The present study tested the use and validity of a stage model of adolescent smoking initiation. The model aims to identify adolescents who are currently not smoking, but who are cognitively predisposed to start smoking in the future. Research on subtypes within the precontemplation stage of adolescent smoking initiation and the concept of susceptibility to smoking led to the construction of a motivational stage model of four distinct stages: committer, immotive, progressive, and contemplator. Using longitudinal data on a large international sample of European adolescents (n=7117), the model proved to have value in predicting smoking initiation at 12 months follow up. The odds ratio to take up regular smoking behavior appeared to double with each forward stage transition. Although effect sizes were small to moderate, unique predictors of transitions from the various stages were identified. Implications of the findings are discussed. PMID- 15135562 TI - Eliciting patients' preferences for cigarette and alcohol cessation: an application of conjoint analysis. AB - The strength and stability of preferences for quitting cigarettes versus alcohol in a population of dual users undergoing treatment was examined using conjoint analysis. Patients at a Veteran's Administration substance abuse treatment center ranked nine vignettes from most to least preferred at baseline and 4 weeks later. The vignettes, using a full factorial design, described health states associated with three levels of substance use. We regressed vignette rankings on the levels of smoking and drinking. A larger regression coefficient indicated a stronger preference for quitting. At baseline and follow-up, the group placed more preference on quitting alcohol than cigarettes (coefficients of 2.23 and 2.35 for alcohol cessation and.51 and.73 for smoking cessation). Some subjects preferred smoking to quitting at baseline (23.9%) and follow-up (23.5%). Over time, 29.4% and 35.3% increased their preference for tobacco and alcohol cessation, while 41.2% and 17.6% decreased their preference for cigarette and alcohol cessation. Preferences for stopping alcohol were stronger than for stopping cigarettes, and many preferences changed after a treatment program. PMID- 15135563 TI - Depression and smoking: from the Transtheoretical Model of change perspective. AB - This study investigated the associations among history of depression, current depressive symptoms, and constructs of the Transtheoretical Model (TTM) of change: stages of change, decisional balance, temptation, and processes of change for smoking cessation. Participants were 239 current smokers (70.7% male, 49.8% African-American, mean cigarettes per day=19.8, 42.3% were in precontemplation). Results showed that participants with a history of depression reported higher current depressive symptoms. No significant relationship was observed between stages and history or current level of depressive symptoms. Current depressive symptoms, rather than depression history, was related to other TTM constructs: pros of smoking, temptation to smoke under habitual and negative affect situations, and the self-reevaluation process of change. PMID- 15135564 TI - The context of sexual risk behavior among heterosexual methamphetamine users. AB - Relatively little is known about the relationship between methamphetamine (meth) use and sexual risk behavior among heterosexual adults. This descriptive study explores the context of sexual risk behavior among HIV-negative, heterosexual meth-using men and women. Drug use history, motivations for meth use, relationship types, and the social-sexual context of meth use were examined as factors that relate to sexual risk behavior and meth use. The majority of participants were white males with an average age of 38.6 years. Primary motivations for current meth use were to get high, to get more energy, and to party. Participants used meth an average of 14 days per month and consumed an average of 7.9 g during this period. The average number of unprotected vaginal, anal, and oral sex acts over a 2-month period were 21.5, 6.3, and 41.7, respectively. Participants reported an average of 9.4 sex partners over 2 months. The findings are discussed in relation to HIV prevention messages and the development of sexual risk reduction interventions for HIV-negative, heterosexual meth users. PMID- 15135565 TI - Thinking about craving: an experimental analysis of smokers' spontaneous self reports of craving. AB - This study evaluated whether smokers generate spontaneous expressions of craving (i.e., expressions of an urge, craving, desire, want, or need) in response to cues designed to provoke a craving state. In a 2 (smoking deprivation: 1 and 12 h) x 2 (cue type: neutral, active) within-subjects design, smokers were asked to think aloud in an unstructured way (i.e., "describe everything you are thinking and feeling right now"). Results revealed a main effect for cue type on think aloud craving responses: Smokers spontaneously generated a greater number of craving-related cognitions during active cue exposure compared with neutral cue exposure, both during both 1- and 12-h deprivation. This same pattern of effects was not found for a self-report assessment of craving, which was insensitive to cue-provoked changes in craving in the 1-h deprivation condition. These results suggest that smokers do spontaneously experience craving, independent of an explicit assessment of craving and that think-aloud methods may provide a novel assessment of craving that may be relatively more sensitive than self-report methods under some circumstances. PMID- 15135566 TI - A comparison of alcohol and drug disorders: is there evidence for a developmental sequence of drug abuse? AB - Whereas the model of Jellinek [Q. J. Stud. Alcohol 7 (1952) 673] of a predictable progression of alcoholism is generally supported, there have been few published studies regarding the natural history of illicit drug disorders. Identification of the development of drug abuse and/or dependence can inform clinicians and researchers on issues, such as diagnosis, prognosis, assessment, and prevention. This study employed a new measure adapted from the 46 events described in Jellinek's progression of alcoholism. Nine licit and illicit substances were also included within the cardsort. Fifty-two individuals motivated to enter drug treatment by a loved one were assessed pretreatment. The drug initiation sequence reported by this sample was as follows: alcohol, tobacco, inhalants, marijuana, and then other drugs. Spearman rank correlation coefficients were conducted between the drug sample and that of Jellinek yielding a modest correlation (r=.35, P=.019). These results suggest that Jellinek's model of progression of alcoholism may also apply to the development of drug disorders. However, important differences found between the alcohol and drug progressions are discussed. PMID- 15135567 TI - Nonclinical panic attack history and smoking cessation: an initial examination. AB - The present study evaluated the association of nonclinical panic attacks among regular smokers with the duration of past quit attempts as well as the type and intensity of DSM-IV smoking withdrawal symptoms. As hypothesized, smokers with a history of panic attacks reported significantly shorter quit attempts compared to their nonpanic counterparts. Additionally, smokers with a history of panic relative to their nonpanic counterparts reported more intense affective reactions during their last quit attempt in regard to anxiety-related but not other types of smoking withdrawal symptomatology. These findings are discussed in regard to the role of negative affect vulnerability factors in smoking cessation with specific reference to panic attacks. PMID- 15135568 TI - Differential contribution of parents and friends to smoking trajectories during adolescence. AB - The purpose of this study was to assess the relative contribution of friends' and parents' smoking on the age of smoking initiation. A sample of 812 preadolescents, who were part of an accelerated longitudinal design, participated in the study over a 4-year period. Three smoking trajectory groups were first established: an age 11-12 starters group (5.7% of the sample), an age 12-13 starters group (11.1% of the sample), and an age 13-14 starters group (7.9% of the sample). A fourth trajectory group included the children who had not started smoking by age 15 years and who represented the majority of the participants (75.4%). After controlling for parental education, gender, and participants' behavioral and academic maladjustment, a series of logistic regressions revealed that parents' smoking assessed during the same year predicted membership in the age 11-12 starters trajectory group. Both parents' and friends' smoking predicted membership in the age 12-13 starters group. Finally, only friends' smoking predicted membership in the age 13-14 starters group. The results are discussed in light of the controversy about the contribution of parents' and friends' smoking behavior to smoking initiation in adolescents. PMID- 15135569 TI - Childhood psychopathology and adolescent cigarette smoking: a prospective survival analysis in children at high risk for substance use disorders. AB - Children of parents with substance use disorders (SUDs) have been shown to demonstrate an increased risk for cigarette smoking in adolescence. In this prospective study, we hypothesized that adolescent cigarette smoking risk would be accounted for by childhood disruptive behavior disorders and parent cigarette smoking. Preadolescent children (ages 10-12 years) of fathers with SUD considered at high average risk (HAR; n=274) and children of fathers without SUD or major psychopathology considered at low average risk (LAR; n=298) participated in structured interviews to determine mental disorder diagnoses and substance use history. Both parents were assessed. The age of onset of daily tobacco use was determined in three follow-up assessments conducted through late adolescence. Conduct disorder (CD) and parental smoking predicted earlier daily cigarette smoking, and mediated the relationship between risk status and offspring daily cigarette smoking. Through the identification of childhood characteristics predicting daily cigarette smoking in adolescence, these results may facilitate targeting of early childhood preventive interventions. PMID- 15135570 TI - A survey on histological image analysis-based assessment of three major biological factors influencing radiotherapy: proliferation, hypoxia and vasculature. AB - Image analysis is a rapidly evolving field with growing applications in science and engineering. In cancer research, it has played a key role in advancing techniques of major diagnostic importance, minimising human intervention and providing vital clinical information. Especially in the field of tissue microscopy, the use of computers for the automated analysis of histological sections is becoming increasingly important. This paper presents an overview of various image analysis methodologies and summarises developments in this field, with great emphasis given on the assessment of three major biological factors known to influence the outcome of radiotherapy: proliferation, vasculature and hypoxia. A brief introduction followed by a survey is provided in each of these areas. PMID- 15135571 TI - Robust and real-time torsional eye position calculation using a template-matching technique. AB - Computation of eye rotation about the line of sight (torsion) using image processing techniques has traditionally used cross-correlation of iral signatures sampled from circular arcs centered on the pupil. We have developed a new algorithm that utilizes a template-matching technique to calculate torsional eye position. Iral signatures are obtained from two annuli centered on the pupil center. By assuming that torsional rotation of the eye is constrained between successive video frames (<2 degrees), only a small window of the previous reference signature is necessary to determine relative torsional eye displacement. This dramatically reduces the number of pixels needed for computing torsion. This algorithm is considerably faster, attains a higher accuracy, and exhibits considerably less noise than the cross-correlation technique. Running on a 800 MHz Intel-based Dual Processor Pentium III, with a Matrox frame grabber, the system is capable of processing three-dimensional eye position at a rate of 120 frames/s. PMID- 15135572 TI - The study of control methods for the robotic testing system for human musculoskeletal joints. AB - Biomechanical testing of human musculoskeletal joints not only requires qualified testing machines and devices, but also needs an excellent control method to obtain better experimental results. In this paper, we take the human functional spinal unit (FSU) as an example to study how to improve the performance of the robotic testing system. First, the mechanical characteristics of the FSU are described and the simplified model (a rigid body-spring system) for the specimen is given. Because the location of the center of rotation (COR) of the specimen affects the performance of the system, so a comprehensive analysis on the location of the COR is carried out. Furthermore, the performance of the robotic testing system can be also improved through the improvement of the control methods. Two control methods have been proposed, one is the improved hybrid control and the other is fuzzy logic control (FLC). PMID- 15135573 TI - Texture-based approaches for identifying neuro-anatomical structures and electrode tracks. AB - An automated approach to identifying electrode tracks and neuro-anatomical structures (nuclei) was developed using texture attributes of their neuro anatomical stains. The properties that make up the texture features of the nuclei include size, shape and distribution of elemental structures. The electrode tracks are characterized by elongated darkened formations due to gliosis. Based on a Gabor wavelet transform, a texture feature vector was constructed, consisting of localized texture energies along different orientations at different scales. Stained images of brainstem sections in the vestibular nuclei were segmented using partitional clustering in feature space. A metric that computes the location of the tracks relative to the nuclei centers was then implemented. This methodology should be useful for quantifying and automating the procedure by which tracks are localized in anatomical structures. PMID- 15135574 TI - A Web-based cost-effective training tool with possible application to brain injury rehabilitation. AB - Virtual reality (VR) has provoked enormous interest in the medical community. In particular, VR offers therapists new approaches for improving rehabilitation effects. However, most of these VR assistant tools are not very portable, extensible or economical. Due to the vast amount of 3D data, they are not suitable for Internet transfer. Furthermore, in order to run these VR systems smoothly, special hardware devices are needed. As a result, existing VR assistant tools tend to be available in hospitals but not in patients' homes. To overcome these disadvantages, as a case study, this paper proposes a Web-based Virtual Ticket Machine, called WBVTM, using VRML [VRML Consortium, The Virtual Reality Modeling Language: International Standard ISO/IEC DIS 14772-1, 1997, available at ], Java and EAI (External Authoring Interface) [Silicon Graphics, Inc., The External Authoring Interface (EAI), available at ], to help people with acquired brain injury (ABI) to relearn basic living skills at home at a low cost. As these technologies are open standard and feature usability on the Internet, WBVTM achieves the goals of portability, easy accessibility and cost-effectiveness. PMID- 15135575 TI - MobileNurse: hand-held information system for point of nursing care. AB - Healthcare information travels with patients and clinicians and therefore the need for information to be ubiquitously available is key to reliable patient care and reliable medical systems. We have implemented MobileNurse, a prototype point of-care system using PDA. MobileNurse has four modules each of which performs: (1) patient information management; (2) medical order check; (3) nursing recording; and (4) nursing care plan. MobileNurse provides easy input interface and various outputs for nursing records. The system consists of PDAs and a mobile support system (MSS) which supports clinical data exchange between PDAs and hospital information system. Two synchronization modules have been developed to keep the patient data consistent between PDAs and MSS. Clinical trials were performed with six volunteered nurses. They tried MobileNurse for 1-day caring simulated patients. According to the survey after the trials, most of volunteers agreed that MobileNurse is more helpful and convenient than other non-mobile care systems to check medical orders and retrieve the results of recent clinical tests at the bedside. Through the involvement, we found out that ease-to-use interface is the most critical successful factor for mobile patient care systems. PMID- 15135577 TI - A computer program to estimate power and relative efficiency to assess multiplicative interactions in flexibly matched case-control studies. AB - Matching on one of two possibly interacting exposures can increase power and efficiency to estimate multiplicative interactions in case-control studies. We recently introduced the concept of flexible matching strategies with varying proportions of a dichotomous matching factor among controls to further increase power and efficiency. In order to facilitate the application of this concept, we developed a computer program which provides estimates of power and efficiency varying the proportion of the matching factor in controls over all possible values from 1 to 99%. The program allows one to estimate the effect of frequency matching on power and efficiency to study multiplicative interactions and to assess the optimal prevalence of the matching factor in selected controls for a given scenario which often differs from the prevalence in cases (aimed at in traditional 'fixed' frequency matching). Our program will strongly facilitate assessing the benefits of matching and flexible matching strategies in case control studies addressing multiplicative interactions, including gene environment interactions. PMID- 15135576 TI - Mixed models for longitudinal left-censored repeated measures. AB - Longitudinal studies could be complicated by left-censored repeated measures. For example, in Human Immunodeficiency Virus infection, there is a detection limit of the assay used to quantify the plasma viral load. Simple imputation of the limit of the detection or of half of this limit for left-censored measures biases estimations and their standard errors. In this paper, we review two likelihood based methods proposed to handle left-censoring of the outcome in linear mixed model. We show how to fit these models using SAS Proc NLMIXED and we compare this tool with other programs. Indications and limitations of the programs are discussed and an example in the field of HIV infection is shown. PMID- 15135578 TI - Growth of pulsed electric field exposed Escherichia coli in relation to inactivation and environmental factors. AB - Pulsed electric fields (PEF) have been proven to inactivate microorganisms during nonthermal conditions and have the potential to replace thermal processing as a method for food preservation. However, there is a need to understand the recovery and growth of survivors and potentially injured microorganisms following PEF processing. The purpose of this investigation was to study the growth of Escherichia coli at 10 degrees C following exposure to electrical field strengths (15, 22.5 and 30 kV/cm) in relation to inactivation and the amount of potentially sublethally injured cells. One medium was used as both a treatment medium and an incubation medium, to study the influence of environmental factors on the inactivation and the growth of the surviving population. The pH (5.0, 6.0 and 7.0) and water activity (1.00, 0.985 and 0.97) of the medium was varied by adding HCl and glycerol, respectively. Growth was followed continuously by measuring the optical density. The time-to-detection (td) and the maximum specific growth rate (micromax) were calculated from these data. Results showed that the PEF process did not cause any obvious sublethal injury to the E. coli cells. The number of survivors was a consequence of the combination of electrical field strength and environmental factors, with pH being the most prominent. Interestingly, the micromax of subsequent growth was influenced by the applied electrical field strength during the process, with an increased micromax at more intense electrical field strengths. In addition, the micromax was also influenced by the pH and water activity. The td, which could theoretically be considered as an increase in shelf life, was found to depend on a complex correlation between electrical field strength, pH and water activity. That could be explained by the fact that the td is a combination of the number of survivors, the recovery of sublethal injured cells and the growth rate of the survivors. PMID- 15135579 TI - Fine-tuning Food Safety Objectives and risk assessment. AB - Food Safety Objectives (FSOs) have been proposed as a practical tool to translate public health targets for food safety into tolerable levels of pathogens in a food product. The FSO concept is subject to intensive debate, and has not been developed in detail. We evaluate the proposed definition of FSOs and their implementation from the perspective of Quantitative Microbiological Risk Assessment (QMRA). The relationship between QMRA and FSOs is illustrated by a model for the public health risk of Shiga-producing Escherichia coli in steak tartare. We conclude that the proposed definition of FSOs needs to be modified to properly account for variability in and uncertainty about the contamination of food with pathogenic microorganisms and emphasize that both prevalence and concentration of pathogens must be considered. For this purpose, we propose the P D equivalence curve, a simple graphical tool to separate "tolerable" from "non tolerable" combinations of prevalence and concentration (dose). PMID- 15135580 TI - Analysis of population structure of Aspergillus flavus from peanut based on vegetative compatibility, geographic origin, mycotoxin and sclerotia production. AB - Isolates of Aspergillus flavus obtained from a new growing peanut region in Argentina (Formosa province) were examined for aflatoxin types B and G and cyclopiazonic acid (CPA) production. Sclerotia diameters and the number of sclerotia produced per square centimetre were also determined for each isolate. They were tested by vegetative compatibility group analysis to investigate their genetic relatedness and correlate the results with vegetative compatibility groups previously described from the major peanut-growing area (Cordoba province) in our country. Two isolates were considered atypical because they simultaneously produce aflatoxins B and G and CPA. A. flavus population from Formosa province was very diverse genetically. Vegetative compatibility groups (VCGs) formed by typical isolations of A. flavus were different among agroecological sites. Formosa isolates could not be grouped to any of the Cordoba VCGs, while that one of the VCGs that contain atypical isolates included strains from the two geographical regions. Each VCG included isolates of the same mycotoxin and sclerotia production pattern. The two regions analysed have different climatic conditions, soil type, crop sequence history and also are in different latitude. These parameters may reflect different geographic adaptation between isolates from both sites. PMID- 15135581 TI - Mould contamination in production of semi-hard cheese. AB - Air, equipment, plastic film, brine and milk were sampled from four cheese factories in Norway during the period September 1997 to May 1999 in order to identify the critical points for mould contamination in the production process. Altogether, 672 samples were collected. Penicillium brevicompactum was the most frequently isolated species from three of the factories, while Geotrichum candidum was the most frequently isolated species from the fourth. P. commune, P. palitans, P. solitum and P. roqueforti ss. roqueforti, all common contaminants on cheese, were found in samples of air and equipment, and the former was also isolated from plastic film. The results in the present study showed that the mould levels in the cheese factories varied between the different control points. The mould levels at some of the air control points had high mould counts while the mould levels in milk and brine, on equipment and on plastic film, generally were low. The fungi at the control points with high mould levels consisted of common cheese contaminants as well as species not commonly isolated from cheese. The statistical analysis showed that air was the major source of the important cheese contaminants P. commune and P. palitans during the production process. High quality air with low number of cheese contaminants in production rooms, especially the wrapping room, is important in order to reduce mould contamination. PMID- 15135582 TI - Alternaria and Fusarium in Norwegian grains of reduced quality--a matched pair sample study. AB - The occurrence and geographic distribution of species belonging to the genera Alternaria and Fusarium in grains of reduced and of acceptable quality were studied post-harvest in 1997 and 1998. A total of 260 grain samples of wheat, barley and oats was analysed. The distribution of Alternaria and Fusarium spp. varied significantly in samples of reduced quality compared with acceptable samples. Alternaria spp. dominated in the acceptable samples with A. infectoria group as the most frequently isolated and most abundant species group of this genus while Fusarium spp. dominated in samples of reduced quality. The most frequently isolated Fusarium spp. from all samples were F. avenaceum, F. poae, F. culmorum and F. tricinctum. Other important toxigenic Fusarium spp. isolated were F. graminearum and F. equiseti. The infection levels of F. graminearum and F. culmorum were significantly higher in the samples of reduced quality. The results indicated a negative interaction between F. graminearum and Alternaria spp. as well as between F. graminearum and other Fusarium spp. PMID- 15135583 TI - Binding interaction studies of the immobilized Salmonella typhimurium with extracellular matrix and muscle proteins, and polysaccharides. AB - Our research attempts to understand the real-time interactions of immobilized Salmonella typhimurium with extracellular membrane proteins (collagen I, fibronectin and laminin) and muscle proteins (actin and myosin). Salmonella cells were immobilized on the sensor chip of a surface plasmon resonance (SPR) biosensor. Typical results showed that collagen I and myosin had higher binding responses to the S. typhimurium surface but laminin, actin and fibronectin had lower binding responses. The binding kinetics of collagen I and Salmonella cell surface showed an apparent dissociation and association rate constants of 3.90 E 4 s(-1) and 1.07 E+4 mol(-1) s(-1). Using the model system developed in our laboratory, the interactions of carrageenans and other polysaccharides with collagen and the Salmonella sensor surface were evaluated. The kappa-carrageenans blocked 92-100% binding of collagen to the Salmonella surface, while sodium alginate and low methoxy pectin blocked 50% and 18% binding, respectively. These biosensor studies allowed the rapid evaluation of compounds that may prevent bacterial attachment to poultry skin and carcasses, thus reducing pathogen contamination of poultry foods. PMID- 15135584 TI - Modelling effect of physical and chemical parameters on heat inactivation kinetics of hepatitis A virus in a fruit model system. AB - While thermal destruction of pathogenic bacteria has been thoroughly studied in food industry, heat inactivation of viruses in food has been poorly investigated. Experiments were carried out to characterize the effects of controlled physical and chemical characteristics of a food matrix upon heat resistance parameters (D and z values) of hepatitis A virus (HAV), taken as model because of its reported heat resistance. Sucrose content (28-52 degrees Brix), calcium concentration (90 1700 mg kg(-1)) and pH (3.3-4.3) were selected for possible influence on thermal inactivation of HAV in strawberry mashes and thus included in an experimental design according to a Doehlert matrix. Use of this design not only allowed to detect and quantify the direct influence of sucrose concentration upon the D85 degrees C value to be higher than the one of pH, but also to reveal a sucrose concentration/pH specific interaction, while no effect of calcium concentration was evidenced. Although the model cannot be directly used to predict heat resistance in real fruit systems, because of differences observed between predicted and measured D85 degrees C values, it is useful for predicting the trends and relative changes in D values due to sucrose concentration and pH variations. Results suggested possible effects of other constituents of strawberry products on heat resistance of HAV and confirmed the importance of experimental validation of any model-derived process. Nevertheless, such a modelling approach using response surface methodology provides a rapid answer to heat resistance evaluation of a food-borne virus as a function of specific physical and chemical parameters of specific food products. PMID- 15135585 TI - Screening of glutamate decarboxylase activity and bile salt resistance of human asymptomatic carriage, clinical, food, and environmental isolates of Listeria monocytogenes. AB - Following consumption, stomach acidity is the first major barrier encountered by the food-borne pathogen Listeria monocytogenes. Analysis of low pH sensitivity and glutamate decarboxylase (GAD) acid resistance system of 14 isolates of L. monocytogenes carried asymptomatically by humans showed that levels of GAD activity were subjected to strain variation. Similar variations were observed for strains responsible for 18 cases of listeriosis, whereas in comparison, 13 strains isolated from food and food-processing plant environments showed lower GAD activity. Following survival of the stomach barrier, L. monocytogenes also has to resist bile salts encountered in the small intestines. Analysis revealed that all strains tested were able to grow in the presence of bile salts with concentrations as high as those encountered in the small intestines and had previously identified bile salt hydrolase (BSH) activity. Strain variation was observed but there was no relationship between the origin of the strains and the ability to degrade bile salts. PMID- 15135587 TI - H+-ATPase activity in Bifidobacterium with special reference to acid tolerance. AB - The acid tolerance of 17 strains of nine species of bifidobacteria was compared using brief exposures to acidic conditions (pH 2-5). In addition, because it has been hypothesized that the acid tolerance of bifidobacteria depends on H+-ATPase activity, the activity of this enzyme in various strains and species was compared. In general, the acid tolerance of bifidobacteria was found to be weak, with the exception of Bifidobacterium lactis and Bifidobacterium animalis. High numbers of all strains of B. lactis and B. animalis survived exposure to pH 3-5 for 3 h. The H+-ATPase activity of the acid-tolerant strains B. lactis LKM512 and JCM 10602T, and B. animalis JCM 1190T, JCM 1253, JCM 7117, and JCM 7124 was higher at pH 4 than at pH 5. In contrast, the H+-ATPase activity of nonacid tolerant strains was lower at pH 4 than at pH 5. PMID- 15135586 TI - PCR-restriction fragment length analysis of aflR gene for differentiation and detection of Aspergillus flavus and Aspergillus parasiticus in maize. AB - Contamination of food and feedstuffs by Aspergillus species and their toxic metabolites is a serious problem as they have adverse effects on human and animal health. Hence, food contamination monitoring is an important activity, which gives information on the level and type of contamination. A PCR-based method of detection of Aspergillus species was developed in spiked samples of sterile maize flour. Gene-specific primers were designed to target aflR gene, and restriction fragment length polymorphism (RFLP) of the PCR product was done to differentiate Aspergillus flavus and Aspergillus parasiticus. Sterile maize flour was inoculated separately with A. flavus and A. parasiticus, each at several spore concentrations. Positive results were obtained only after 12-h incubation in enriched media, with extracts of maize inoculated with A. flavus (101 spores/g) and A. parasiticus (104 spores/g). PCR products were subjected to restriction endonuclease (HincII and PvuII) analysis to look for RFLPs. PCR-RFLP patterns obtained with these two enzymes showed enough differences to distinguish A. flavus and A. parasiticus. This approach of differentiating these two species would be simpler, less costly and quicker than conventional sequencing of PCR products. PMID- 15135588 TI - Souring and breakdown of cyanogenic glucosides during the processing of cassava into akyeke. AB - The population and composition of the lactic acid bacteria microbiota as well as the content of cyanogenic glucosides occurring at various stages of fermentation and subsequent processing of cassava roots into akyeke, a steamed sour cassava meal, were investigated. The number of lactic acid bacteria and percentage titratable acidity increased during 5 days of fermentation, but decreases were observed in the subsequent operations of 'washing' the dough with water followed by partial drying and steaming. In field and laboratory samples, Lactobacillus plantarum accounted for 59.3% and 52.3%, Lactobacillus brevis 23.3% and 22.8% and Leuconostoc mesenteroides subsp. cremoris 14.5% and 15.8%, respectively, of all lactic acid bacteria isolated at various stages of fermentation and processing. A reduction of about 98% occurred in the total cyanogens (CN) content of cassava roots during processing, from 69.3 to 1.4 and 110.3 to 2.8 mg CN equivalent/kg dry weight for laboratory and field samples of akyeke, respectively. PMID- 15135589 TI - Educating, with evidence. PMID- 15135590 TI - Carotid endarterectomy. PMID- 15135591 TI - Diagnosis of lung rejection. PMID- 15135592 TI - A causal role for human papillomavirus in head and neck cancer. PMID- 15135593 TI - The unkindest cup. PMID- 15135594 TI - Prevention of disabling and fatal strokes by successful carotid endarterectomy in patients without recent neurological symptoms: randomised controlled trial. AB - BACKGROUND: Among patients with substantial carotid artery narrowing but no recent neurological symptom (stroke or transient ischaemia), the balance of surgical risks and long-term benefits from carotid endarterectomy (CEA) was unclear. METHODS: During 1993-2003, 3120 asymptomatic patients with substantial carotid narrowing were randomised equally between immediate CEA (half got CEA by 1 month, 88% by 1 year) and indefinite deferral of any CEA (only 4% per year got CEA) and were followed for up to 5 years (mean 3.4 years). Kaplan-Meier analyses of 5-year risks are by allocated treatment. FINDINGS: The risk of stroke or death within 30 days of CEA was 3.1% (95% CI 2.3-4.1). Comparing all patients allocated immediate CEA versus all allocated deferral, but excluding such perioperative events, the 5-year stroke risks were 3.8% versus 11% (gain 7.2% [95% CI 5.0-9.4], p<0.0001). This gain chiefly involved carotid territory ischaemic strokes (2.7% vs 9.5%; gain 6.8% [4.8-8.8], p<0.0001), of which half were disabling or fatal (1.6% vs 5.3%; gain 3.7% [2.1-5.2], p<0.0001), as were half the perioperative strokes. Combining the perioperative events and the non-perioperative strokes, net 5-year risks were 6.4% versus 11.8% for all strokes (net gain 5.4% [3.0-7.8], p<0.0001), 3.5% versus 6.1% for fatal or disabling strokes (net gain 2.5% [0.8 4.3], p=0.004), and 2.1% versus 4.2% just for fatal strokes (net gain 2.1% [0.6 3.6], p=0.006). Subgroup-specific analyses found no significant heterogeneity in the perioperative hazards or (apart from the importance of cholesterol) in the long-term postoperative benefits. These benefits were separately significant for males and females; for those with about 70%, 80%, and 90% carotid artery narrowing on ultrasound; and for those younger than 65 and 65-74 years of age (though not for older patients, half of whom die within 5 years from unrelated causes). Full compliance with allocation to immediate CEA or deferral would, in expectation, have produced slightly bigger differences in the numbers operated on, and hence in the net 5-year benefits. The 10-year benefits are not yet known. INTERPRETATION: In asymptomatic patients younger than 75 years of age with carotid diameter reduction about 70% or more on ultrasound (many of whom were on aspirin, antihypertensive, and, in recent years, statin therapy), immediate CEA halved the net 5-year stroke risk from about 12% to about 6% (including the 3% perioperative hazard). Half this 5-year benefit involved disabling or fatal strokes. But, outside trials, inappropriate selection of patients or poor surgery could obviate such benefits. PMID- 15135595 TI - I never did become a cardiologist. PMID- 15135596 TI - Prediction of lung-transplant rejection by hepatocyte growth factor. AB - BACKGROUND: Graft rejection is a major complication of lung transplantation. No serological marker of rejection is in common use. Hepatocyte growth factor (HGF) is highly expressed in the lung and produced after acute lung injury; serum concentrations increase in inflammatory lung diseases. We investigated whether HGF could be an accurate marker for prediction of lung-graft rejection. METHODS: Serum concentrations of HGF were measured by ELISA in 109 patients who had undergone lung transplantation (65 for chronic obstructive pulmonary disease; 23 for cystic fibrosis; 21 for idiopathic lung fibrosis), comparing those who had no subsequent events and those with episodes of infection or rejection, as well as in 12 healthy controls. FINDINGS: The mean baseline serum HGF concentration was 645 ng/L (SD 259) in controls and 1358 ng/L (603) in the patients before transplantation. After transplantation the mean concentration in patients with no events was 1147 ng/L (510) compared with 1559 ng/L (323) in patients with infection (p=0.001 vs controls; change from pretransplant value not significant). Patients with rejection had significantly higher concentrations than all other groups (3972 ng/L [1463], p<0.0001). Logistic regression identified HGF as a predictor for lung graft rejection (p=0.012). After steroid treatment, HGF concentrations returned almost to the preoperative values within 3 days. INTERPRETATION: HGF might be a marker for graft rejection in lung transplantation. A potential link between viral infection, mainly cytomegalovirus, and HGF, however, remains to be investigated. PMID- 15135597 TI - Burdens of disease in southern Africa. PMID- 15135599 TI - Association between microfilarial load and excess mortality in onchocerciasis: an epidemiological study. AB - BACKGROUND: Infection with the parasitic filarial nematode Onchocerca volvulus can lead to severe visual impairment and ultimately blindness. Excess mortality has been noted among people with onchocerciasis, but it is not clear whether this effect is entirely due to blindness, or mediated by some more direct effects of the infection. METHODS: We assessed the relations between infection with O volvulus, visual acuity, and host mortality with data obtained by the Onchocerciasis Control Programme in West Africa from 2315 villages in 11 countries. FINDINGS: 297,756 people were eligible for follow-up, and accumulated 2,579449 person-years of follow-up from 1971 through 2001. 24,517 people died during this period; 1283 (5.2%) of these deaths were due to onchocerciasis. Mortality of the human host was significantly and positively associated with increasing microfilarial burden (p<0.00001), but not with blindness after adjustment for microfilarial load and other variables. Overall, after adjustment for microfilarial load and other variables, female individuals had a risk of death about 7.5% lower than males (p<0.00001). Rates of mortality peaked in the mid 1980s but generally decreased thereafter. INTERPRETATION: We have shown a direct relation between O volvulus microfilarial load and host mortality in a comprehensive dataset and in both sexes. PMID- 15135598 TI - Failure to clear persistent vaccine-derived neurovirulent poliovirus infection in an immunodeficient man. AB - BACKGROUND: Individuals who chronically excrete neurovirulent poliovirus of vaccine-origin are of considerable concern to the Global Polio Eradication programme. Chronic infection with such polioviruses is a recognised complication of hypogammaglobulinaemia. METHODS: We did a series of in-vitro and in-vivo therapeutic studies, with a view to clearing persistent neurovirulent poliovirus infection in an individual with common variable immunodeficiency, using oral immunoglobulin, breast milk (as a source of secretory IgA), ribavirin, and the anti-picornaviral agent pleconaril. We undertook viral quantitation, antibody neutralisation and drug susceptibility assays, and viral gene sequencing. FINDINGS: Long-term asymptomatic excretion of vaccine-derived neurovirulent poliovirus 2 was identified in this hypogammaglobulinaemic man, and was estimated to have persisted for up to 22 years. Despite demonstrable in-vitro neutralising activity of immunoglobulin and breast milk, and in-vitro antiviral activity of ribavirin, no treatment was successful at clearing the virus, although in one trial breast milk significantly reduced excretion levels temporarily. During the course of study, the virus developed reduced susceptibility to pleconaril, precluding the in-vivo use of this drug. Sequence analysis revealed the emergence of a methionine to leucine mutation adjacent to the likely binding site of pleconaril in these isolates. INTERPRETATION: Chronic vaccine-associated poliovirus infection in hypogammaglobulinaemia is a difficult condition to treat. It represents a risk to the strategy to discontinue polio vaccination once global eradication has been achieved. PMID- 15135600 TI - An unusual cause of psychosis. PMID- 15135601 TI - Histo-blood group type change of the graft from B to O after ABO mismatched heart transplantation. AB - A patient with blood group type O accidentally received a blood group B cardiac allograft. He died almost 5 years after transplantation because of transplant vasculopathy. We monitored the expression of ABO-type antigens on the vascular endothelial cells of the cardiac allograft for 44 months, using monoclonal antibodies for immunohistology. The antigenic profile of the graft endothelial cells changed progressively from B to O. This change was first detectable 14 months after transplantation and it was most prominent at the end of the observation period. Thus, we have shown that the allograft blood type of the endothelium of this patient's heart began to change from type B to his own, 1 year after transplantation. By 44 months post-transplantation, it had changed to O type. PMID- 15135602 TI - Assessment of NICE guidance on two surgical procedures. AB - The National Institute of Excellence (NICE) released its first guidance in 2000 on clinical procedures and technologies in England and Wales. Using hospital activity data, we looked at the effect of NICE guidance on wisdom tooth extraction and primary total hip replacement in the UK National Health Service. We selected hospitals that submitted their activity continuously between 1997 and 2002. Analysis of wisdom tooth extraction showed that NICE guidance was not the primary reason for the downward trend in extractions (p=0.8287); this trend was more likely to be attributable to previous guidance. For total hip replacement there was no significant change in behaviour away from use of non-cemented hip prostheses. Therefore, NICE guidance has had little effect on behaviour for these two procedures. PMID- 15135603 TI - Meningiomas. AB - Meningiomas are by far the most common tumours arising from the meninges. Progressive enlargement of the tumour leads to focal or generalised seizure disorders or neurological deficits caused by compression of adjacent neural tissue. Surgery remains the primary treatment of choice, although the use of fractionated radiotherapy or stereotactic single-dose radiosurgery is increasing for meningiomas that are incompletely excised, surgically inaccessible, or recurrent and either atypical or anaplastic. Although most meningiomas have good long-term prognosis after treatment, there are still controversies over management in a proportion of cases. We review various features of meningioma biology, diagnosis, and treatment and provide an overview of the current rationale and evidence base for the various therapeutic approaches. PMID- 15135604 TI - Autoantibodies as predictors of disease. AB - CONTEXT: Many human diseases are the result of autoimmune attack, presumably related to a loss of tolerance to self. Autoimmune disease can be divided into either organ-specific illnesses, such as thyroid disease, type 1 diabetes, and mysasthenia gravis, or systemic illnesses, such as rheumatoid arthritis and systemic lupus erythematosus. The pathogenesis of autoimmune damage also segregates autoimmune disease in that some diseases or manifestations are mainly induced by autoantibodies. Pathogenesis may be mainly mediated by autoimmune T lymphocytes. Notwithstanding the underlying mechanism of disease, virtually all autoimmune diseases are associated with circulating autoantibodies, which bind self-protein. Furthermore, for many diseases these autoantibodies are found in serum samples many years before disease onset. STARTING POINT: In the past several years a new autoantibody specificity has been identified in the sera of patients with rheumatoid arthritis. These autoantibodies bind citrulline, a post translational modification of arginine. Markus Nielen and colleagues recently studied the presence of these autoantibodies and rheumatoid factor in blood donors who later developed rheumatoid arthritis (Arthritis Rheum 2004; 50: 380 86). About half the patients were positive for at least autoantibody at a median of 4.5 years before the onset of disease. The negative predictive value of these tests was high, while the positive predictive value was very high. WHERE NEXT? Autoantibodies might not be directly responsible for many of the manifestations of autoimmune disease, but they are markers of future disease in presently healthy individuals. Long-term large studies of outcome are needed to assess the use of assaying autoantibodies for prediction of disease. Such data could lead to intervention trials to prevent autoimmune disease, as are already underway in type 1 diabetes. PMID- 15135605 TI - The future of rotavirus vaccines: a major setback leads to new opportunities. PMID- 15135606 TI - Catastrophic failures of public health. PMID- 15135607 TI - Catastrophic failures of public health. PMID- 15135608 TI - Catastrophic failures of public health. PMID- 15135609 TI - Catastrophic failures of public health. PMID- 15135610 TI - Catastrophic failures of public health. PMID- 15135611 TI - Catastrophic failures of public health. PMID- 15135612 TI - Timing of surgery for symptomatic carotid stenosis. PMID- 15135613 TI - A network of excellence. PMID- 15135614 TI - A network of excellence. PMID- 15135615 TI - Procalcitonin-guided antibiotic treatment in heart failure. PMID- 15135617 TI - Fetal alcohol syndrome. PMID- 15135618 TI - Attitudes towards delivering bad news in Peru. PMID- 15135619 TI - Renal-cell carcinoma: vaccination and risk of tumour progression. PMID- 15135621 TI - "New" practice of bedsharing and risk of SIDS. PMID- 15135622 TI - Thyroxine adherence in primary hypothyroidism. PMID- 15135623 TI - Gregory Stock. PMID- 15135627 TI - Lineage commitment and developmental plasticity in early lymphoid progenitor subsets. PMID- 15135628 TI - The CD4/CD8 lineage choice: new insights into epigenetic regulation during T cell development. PMID- 15135629 TI - CD4/CD8 coreceptors in thymocyte development, selection, and lineage commitment: analysis of the CD4/CD8 lineage decision. PMID- 15135630 TI - Development and function of T helper 1 cells. PMID- 15135632 TI - Generation, maintenance, and function of memory T cells. PMID- 15135631 TI - Th2 cells: orchestrating barrier immunity. PMID- 15135633 TI - CD8+ effector cells. PMID- 15135634 TI - An integrated model of immunoregulation mediated by regulatory T cell subsets. PMID- 15135636 TI - Cancer-associated genes can affect somatic intrachromosomal recombination early in carcinogenesis. AB - The pKZ1 recombination mutagenesis model has provided a sensitive assay where we study somatic intrachromosomal recombination (SICR) as a mutation end-point. SICR is associated with non-homologous end-joining repair of double-strand breaks and can result in chromosomal inversions and deletions, both of which are common chromosomal aberrations identified in cancers. It has been difficult to study the effect of cancer-associated genes on chromosomal changes prior to tumour formation in vivo because of a lack of appropriate test systems. We hypothesised that cancer-associated genes play a role in formation of chromosomal aberrations and that the pKZ1 model would provide a system in which such a role could be studied in the initial steps of carcinogenesis. Transgenic tumour model mice were bred to pKZ1 mice to produce double transgenic animals. SICR inversion events were scored in mouse tissues at an early time, prior to evident tumour formation, and compared with endogenous pKZ1 SICR levels. Over-expression of the c-myc proto oncogene resulted in a significant 2.1-fold increase in SICR in spleen. Loss of Msh2 and expression of the SV40 T antigen resulted in a significantly reduced SICR frequency (0.3 of the endogenous frequency in pKZ1 mice) in spleen and prostate respectively. Therefore SICR was affected in the case of all three cancer-associated genes studied. We hypothesise that the increase and decrease in SICR in the presence of cancer-associated genes results from incorrect repairing of double-strand breaks. The data presented here suggest that the pKZ1 model may provide a powerful tool for studying the effect of cancer-associated genes on chromosomal changes in the early stages of carcinogenesis. PMID- 15135637 TI - Zinc finger proteins and other transcription regulators as response proteins in benzo[a]pyrene exposed cells. AB - Proteomic analysis, which combines two-dimensional electrophoresis (2-DE) and mass spectrometry (MS), is an important approach to screen proteins responsive to specific stimuli. Benzo[a]pyrene (B[a]P), a prototype of polycyclic hydrocarbons (PAHs), is a potent procarcinogen generated from the combustion of fossil fuel and cigarette smoke. To further probe the molecular mechanism of mutagenesis and carcinogenesis, and to find potential molecular markers involved in cellular responses to B[a]P exposure, we performed proteomic analysis of whole cellular proteins in human amnion epithelial cells after B[a]P-treatment. Image visualization and statistical analysis indicated that more than 40 proteins showed significant changes following B[a]P-treatment (P < 0.05). Among them, 20 proteins existed only in the control groups, while six were only present in B[a]P treated cells. In addition, the expression of 10 proteins increased whereas 11 decreased after B[a]P-treatment. These proteins were subjected to in-gel tryptic digestion followed by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) analysis. Using peptide mass fingerprinting (PMF) to search the nrNCBI database, we identified 22 proteins. Most of these proteins have unknown functions and have not been previously connected to a response to B[a]P exposure. To further annotate the characteristics of these proteins, GOblet analysis was carried out and results indicated that they were involved in multiple biological processes including regulation of transcription, cell proliferation, cell aging and other processes. However, expression changes were noted in a number of transcription regulators, including eight zinc finger proteins as well as SNF2L1 (SWI/SNF related, matrix associated, actin dependent regulator of chromatin, subfamily a, member 1), which is closely linked to the chromatin remodeling process. These data may provide new clues to further understand the implication of these proteins in cellular responses to carcinogen exposure as well as the molecular mechanisms of B[a]P-induced mutagenesis and carcinogenesis. PMID- 15135638 TI - Mechanism of 2-aminopurine-stimulated mutagenesis in Escherichia coli. AB - 2-Aminopurine (2AP), a base analog, causes both transition and frameshift mutations in Escherichia coli. The analog is thought to cause mutations by two mechanisms: directly, by mispairing with cytosine, and indirectly, by saturation of mismatch repair (MMR). The goal of this work was to measure the relative contribution of these two mechanisms to the occurrence of transition mutations. Our data suggest that, in contrast to 2-aminopurine-stimulated frameshift mutations, the majority of transition mutations are a direct effect of base mispairing. PMID- 15135639 TI - Long-chain adducts of trans-4-hydroxy-2-nonenal to DNA bases cause recombination, base substitutions and frameshift mutations in M13 phage. AB - Oxidative stress enhances lipid peroxidation (LPO) implicated in the promotion and progression of carcinogenesis. One of the major LPO products is trans-4 hydroxy-2-nonenal (HNE), which was shown to react with guanosine and under peroxidizing conditions also with adenosine. We show here that all four DNA bases are targets for HNE, although displaying different reactivity: dG > dC > dA approximately equal to dT. HPLC and mass spectrometry analyses of HNE reactions with deoxynucleosides showed in each case the formation of several products, with mass peaks corresponding to HNE-dN adducts at a 1:1 and also 2:1 and 3:1 ratios. In the dA, dC and dG reactions, mass peaks corresponding to heptyl-substituted etheno-adducts were also detected, indicating HNE oxidation to its epoxide by air oxygen. In DNA pretreated with HNE, DNA synthesis by T7 DNA polymerase was stopped in a sequence-dependent manner at G > or = C > A and T sites. HNE increased the mutation rates in the lac Z gene of M13 phage transfected into wild type Escherichia coli. The most frequent event was the recombination between lacZ gene sequences in M13 and the E. coli F' factor DNA. Base substitutions and frameshifts were also observed in approximately similar numbers. Over 50% of base substitutions were the C-->T transitions, followed by the G-->C and A-->C transversions. In the E. coli recA strain recombination was not observed, although one mutational G-->T hot-spot appeared within the DNA fragment undergoing recombination in the wild type E. coli. We conclude that long chain HNE adducts to DNA bases arrest DNA synthesis and cause recombination, base substitutions and frameshift mutations in ssDNA. PMID- 15135640 TI - Response of REV3 promoter to N-methyl-N'-nitro-N-nitrosoguanidine. AB - Previously, we have shown that low concentration of N-methyl-N'-nitro-N nitrosoguanidine (MNNG) led to the upregulation of REV3 gene at transcriptional level in cultured human amnion FL cells. In this study, using bioinformatic analysis the putative binding sites for different transcription factors were found to exist in REV3 gene promoter region. A 2570-bp fragment of the 5' flanking region of REV3 gene was amplified by PCR from PAC clone RP3-415N12 and inserted into the pGL3-Basic reporter vector. Dual-luciferase reporter assay demonstrated that the reconstructed plasmid did respond to MNNG exposure in transfected FL cells. Several variants of the reporter plasmids with different deletions of the REV3 promoter region were also constructed and their promoter strength was analyzed. It was found that the MNNG response element might locate at the REV3 gene promoter region -404 to -102 between two Sma1 sites. The shortest responsive fragment containing the putative binding sites for transcription factors CREBP, AP-2, NF-kappaB, and SP1 was also identified. PMID- 15135641 TI - Dissimilar genome response to acute and chronic low-dose radiation in male and female mice. AB - The long-term genetic consequences of chronic exposure to low-dose irradiation constitutes a major concern to the general public and research community, especially as chronic radiation has recently been proven to be much more mutagenic and carcinogenic than previously thought. Here we report the results of the first ever comparison of the effects of acute and chronic whole body low-dose radiation exposure on global gene expression. We found a substantial difference between males and females in the expression of genes involved in signaling, growth control, transcription and other pathways upon acute and chronic radiation exposure. Specifically, we found sex differences in the expression of genes coding for G protein-coupled receptors and nuclear receptors. We also found different induction of PKCdelta, PKCbeta and PKCmu, members of PKC signaling pathway as well as in TGF and WNT signaling in males and females. Very pronounced difference, that was confirmed on the level of protein, was observed in the expression of WNT5A that plays an important role in carcinogenesis and muscle regeneration. WNT5A expression was significantly elevated only in chronically exposed females. We also provide the first evidence of the effect of ionizing radiation on the estrogen receptor in females. Repetitive irradiation of muscle tissue has been linked to development of rhabdomyosarcoma (RMS), which, enigmatically, occurs more frequently in males. Our data may be used to study possible mechanisms of RMS development upon chronic radiation exposure. They may provide some clues about the molecular background of the sex differences of RMS occurrence and may in the future lead to the discovery of new biomarkers for RMS predisposition in the irradiated tissue. Overall, differences in male and female responses to acute and chronic low-dose radiation obtained by this study were more drastic than we could have predicted. If confirmed in other experimental systems, these findings could potentially lead to fundamental changes in radiation safety regulations. PMID- 15135642 TI - Mutational biases associated with potential iron-binding DNA motifs in rodent lacI and human p53 mutational databases. AB - The role of Fenton oxidants in DNA damage, aging, and cancer is appreciated, but not well understood. Six potential iron-binding (PIB) DNA motifs were previously identified as sites of preferential strand cleavage. Since DNA-metal binding domains are a known determinant of oxidative DNA damage, and the location of strand breaks explains where oxidant attack occurs, we sought to determine whether the likelihood of base change mutations is a function of neighboring PIB motifs. We developed a sliding window function that computes the density of PIB motifs on both strands, within 4-12bp, for each location along a target gene. This range of window sizes reflects known diffusion distances of Fenton reaction products. Using mutational databases, odds of mutation at each base were calculated relative to PIB motif density, for all PIB motif types in aggregate, or for individual PIB motifs. Using mutational data from lacI transgenic animals, we observed a non-random distribution of PIB motifs, associated with increased odds of mutation, showing a strand bias. Sensitivity analysis confirmed that the optimum association between PIB motif density and mutations occurs when a 7bp radius is used for the window size. Randomly simulated mutations showed no association with PIB motif density. When the method was applied to human TP53 mutation data, we saw similar results, but no strand bias. As PIB motif density rises, linear trends are observed for increasing odds of mutation. Sensitivity analysis revealed associations between PIB motifs and GC --> AT transitions and GC --> TA transversions-the most commonly observed types of mutations arising from oxidative DNA damage. DNA-metal binding motifs are found in a wide variety of biological contexts, including many where conformational sensitivity to redox state is important. These techniques can help elucidate how DNA-iron-binding may affect lesions and subsequent mutations from multiple agents. PMID- 15135643 TI - No-effect level in the mutagenic activity of the drug cyproterone acetate in rat liver. Part I. Single dose treatment. AB - The anti-androgen and progestagen cyproterone acetate (CPA) is known to cause liver tumors in rats. The drug has been identified recently as a mutagen in the liver of female transgenic lambdalacI (Big Blue) rats at high doses after an expression time of 6 weeks. A dose of 50 mg CPA/kg BW, however, did not increase the mutation frequency (MF) of controls indicating a no-effect level of mutagenicity [Carcinogenesis 19 (1998) 241]. The present study was performed to assess the existence of a no-effect level of mutagenicity. In order to figure out conditions of maximum response, the time course of the MF was determined after administration of a single dose of 100 mg CPA/kg BW to female Big Blue rats. The MF showed a strong initial rise to a maximum 2 weeks after CPA administration accompanied by a corresponding increase of cell proliferation and of DNA adduct levels. Thereafter, the MF decreased within further 2 weeks to one third of the maximum level which was maintained for another 4 weeks. The DNA adduct levels decreased only by 15% during this time period suggesting that mutated hepatocytes were eliminated predominantly. A dose dependence curve determined at a fixation time of 2 weeks revealed a no-effect level of 5 mg CPA/kg BW for mutagenicity. In conclusion, our findings indicate that the length of the observation period may be a critical determinant for the outcome of a mutagenesis study in rat liver. Furthermore, the existence of a no-effect level for the mutagenicity of CPA in rat liver was confirmed. However, it has to be clarified whether the dose of 5 mg CPA/kg BW corresponding to the "transient" type of mutations or the previous dose of 50 mg CPA/kg BW related to a "permanent" type of mutations is more relevant for the assessment of the genotoxic risk. PMID- 15135644 TI - No-effect level in the mutagenic activity of the drug cyproterone acetate in rat liver. Part II. Multiple dose treatment. AB - In order to probe for the existence of a no-effect levels for mutations induced by multiple dose treatment with cyproterone acetate (CPA), female lacI-transgenic Big Blue rats were treated daily for 3 weeks with oral doses of 5.0, 1.0 or 0.2mg/kg CPA b.w., respectively. The dose of 5mg/kg CPA b.w. ineffective as a single dose (see part I) increased the mutation frequency by 2.5-fold. Daily treatment with 1.0 and with 0.2 mg/kg CPA b.w. for 3 weeks, however, was not effective indicating that the 1 mg dose represents a no-effect level for multiple dose treatment. The finding that a total dose of 21 x 5 = 105 mg/kg CPA b.w. caused 50% less DNA adducts, but a mutation frequency three-fold higher (data extrapolated from part I) than observed after daily treatment with 5mg/kg CPA b.w. for 21 days points to a crucial role of cell proliferation in mutagenesis by CPA. Our present results, in combination with previous findings, offer a basis to estimate the risk to develop mutations in human liver following treatment with CPA. Previous studies revealed that CPA-DNA adducts are formed in human hepatocytes at lower levels than in those of female rats [Mutat. Res. 395 (1997) 179; Cancer Res. 56 (1996) 4391]. Moreover, an in vitro-study indicated that human hepatocytes in culture do not respond to the mitogenic effect of CPA, while rat hepatocytes did [Cancer Res. 51 (1991) 1143]. We conclude that the risk of humans to develop mutations under treatment with CPA is substantially lower than in the female rat. PMID- 15135645 TI - Equine estrogen metabolite 4-hydroxyequilenin induces anchorage-independent growth of human mammary epithelial MCF-10A cells: differential gene expression. AB - Long-term exposure to synthetic and endogenous estrogens has been associated with the development of cancer in several tissues. One potential mechanism of estrogen carcinogenesis involves catechol formation and these catechols are further oxidized to electrophilic/redox active o-quinones, which have the potential to both initiate and promote the carcinogenic process. Previously we showed that 4 hydroxyequilenin (4-OHEN) autoxidized to an o-quinone and caused a variety of damage to DNA. Since these deleterious effects could contribute to gene mutations, we investigated the Chinese hamster V79 cells to ascertain the relative ability of estradiol, 4-hydroxyestradiol, 17beta-hydroxyequilenin, 4,17beta-hydroxyequilenin, estrone, 4-hydroxyestrone, equilenin, and 4 hydroxyequilenin to induce the mutation of the hypoxanthine-guanine phosphoribosyltransferase (hprt) gene. All the 4-hydroxylated catechols induced significantly more colony formations in V79 cells as compared to the parent phenols at 100nM, suggesting that the catechol estrogen metabolites are more mutagenic towards the hprt gene than estrogens. Since 4-OHEN induced the highest mutation frequency, we examined a biomarker for transformation potential of this compound in MCF-10A cells using an anchorage-independent growth assay. Although 4 OHEN induced anchorage-independent growth of these cells, the isolated clones were not able to grow as tumors in vivo when injected into nude mice. These cells were assayed for genetic changes using cDNA microarrays. Real time RT-PCR confirmation of some of the differentially expressed genes showed down-regulation of metallothionein 2A, p53, BRCA1, and c-myc. Moreover, we showed the involvement of other genes important in cell transformation and oxidative stress, strengthening the hypothesis that this mechanism plays a considerable role in 4 OHEN-induced anchorage-independent growth. PMID- 15135646 TI - DNA damage in lung after oral exposure to diesel exhaust particles in Big Blue rats. AB - Several chemical mutagens and carcinogens, including polycyclic aromatic hydrocarbons (PAHs) and nitrated PAHs, are adsorbed to the surface of diesel exhaust particles (DEP). DEP can induce formation of reactive oxygen species and cause oxidative DNA damage as well as bulky carcinogen DNA adducts. Lung tissue is a target organ for DEP induced cancer following inhalation. Recent studies have provided evidence that the lung is also a target organ for DNA damage and cancer after oral exposure to other complex mixtures of PAHs. The genotoxic effect of oral administration of DEP was investigated, in terms of markers of DNA damage, mutations and repair, in the lung of Big Blue rats fed a diet with 0, 0.2, 0.8, 2, 8, 20 or 80 mg DEP/kg feed for 21 days. There was no significant increase in the mutation frequency in the cII gene. However, an increase of DNA damage measured as DNA strand breaks (comet assay) and bulky DNA adducts (32P post labeling) was observed. The level of DNA strand breaks increased significantly at all dose levels while the level of DNA adducts increased significantly only at the intermediate dose levels. Similarly, the number of oxidized DNA bases measured as endonuclease III and fapyguanine glycosylase (FPG) sensitive sites increased at the intermediate dose levels. The induction of DNA damage by DEP exposure did not increase the expression of the repair genes OGG1 and ERCC1 at the mRNA level. The present study indicates that the lung is a target organ for primary DNA damage following oral exposure to DEP. DNA damage was induced following exposure to relatively low levels of DEP, but under the conditions used in the present experiment DNA damage did not result in an increased mutation rate. PMID- 15135647 TI - Melphalan-induced DNA damage in p53(+/-) and wild type mice analysed by the comet assay. AB - Melphalan is an alkylating substance used as a therapeutic agent; its mutagenicity is related to its ability to produce monoadducts and to form DNA cross-links. The alkaline comet assay is a useful test for the detection of DNA lesions. However, cross-links are not easily detected under standard conditions. Recently, modifications to the test have been introduced to measure cross-links by evaluating the reduction in induced DNA migration. In this work, the standard comet assay and an assay modified by prolonging the electrophoresis time have been applied to evaluate DNA lesions induced by single, 4 or 26 weekly oral administrations of melphalan to p53(+/-) knockout and to isotype parental mice. Cells were analysed from the liver, bone marrow, peripheral blood and the distal intestine. Moreover, a further protocol in which the presence of cross-links was inferred by the reduction in X-ray-induced DNA migration was applied to bone marrow cells and the sensitivity of the different methods was compared. The majority of groups examined by the standard protocol showed no difference compared to controls, while the modified protocol (prolonged electrophoresis time) could detect a retarded DNA migration in cells from all the organs analysed with the exception of bone marrow cells. Only the protocol based on X-ray in vitro irradiation showed the presence of melphalan-induced cross-links in bone marrow cells exposed to 2mg/kg for 4 weeks, demonstrating that this was the most sensitive approach for detecting this type of lesion. DNA lesions were evident in all the organs analysed. However, results suggest that the kinetics of cross-link repair could be different in bone marrow cells compared to other organs tested. After comparison between genotype-matched treated and control groups, a significant effect was shown more frequently in p53(+/-) than in wild type groups. PMID- 15135648 TI - Non-opioid actions of opioid peptides. AB - Beside the well known actions of opioid peptides on mu-, delta- and kappa-opioid receptors, increasing amount of pharmacological and biochemical evidence has recently been published about non-opioid actions of various opioid peptides. These effects are not abolished by naloxone treatments. Such non-opioid effects are observed both in nervous tissues and in the cellular elements of the immune system. Peptides exhibiting non-opioid effects include beta-endorphin, dynorphin A, nociceptin/OFQ, endomorphins, hemorphins and a number of Proenkephalin A derived peptides, such as Met-enkephalin, Met-enkephalin-Arg-Phe (MERF) and bovine adrenal medullary peptide (BAM22). Non-opioid actions are exerted through different neuronal receptors, e.g., dynorphin hyperalgesia through NMDA receptor, Met-enkephalin induced regulation of cell growth through zeta receptors, pain modulation by nociceptin through ORL-1 or NOP receptors, while BAM22 acts through sensory neuron specific G protein-coupled receptors (SNSR). We have investigated Met-enkephalin-Arg-Phe (MERF) and its analogues by the means of direct and indirect radioligand binding assays. It has been found that in addition to kappa(2) and delta-opioid receptors, MERF can act also through sigma(2)- or probably via FMRF-NH(2) receptors in rat cerebellum. A role of functionally assembling heterodimer receptors in mediating the non-conventional actions of these peptide ligands can not be excluded as well. PMID- 15135649 TI - The peroxisome proliferator BR931 kills FaO cells by p53-dependent apoptosis. AB - Although suppression of apoptosis has been implicated as a mechanism for the hepatocarcinogenicity of peroxisome proliferators (PPs), they can also induce cell death in rat AH130 and human HepG2 hepatoma cells. To study how PPs induce cell death and to characterize the molecular events involved, we administered the hypolipidemic BR931, a peroxisome proliferator, to rat hepatoma FaO cells. Treatment with increasing concentrations of BR931 (0.015 to 0.6 mM) reduced cell viability in a dose- and time-dependent manner, associated with DNA fragmentation and morphological changes characteristic of apoptosis. BR931 also caused phosphorylation of p53 within 3 hours, translocation of the pro-apoptotic Bax protein to mitochondria, release of cytochrome-c into the cytosol, and activation of caspase-9 and -3. These results indicated that BR931 activated the intrinsic caspase cascade. Pretreatment with three different antioxidants, N acetylcysteine, Vitamin C and Trolox, reduced apoptosis, suggesting that reactive oxygen species (ROS) plays a role in BR931-induced apoptosis. In support of this hypothesis, BR931 produced increased levels of 8-hydroxy-deoxy-guanosine, a marker of DNA oxidative damage. Antioxidants prevented the p53 phosphorylation, up-regulation of Bax and BR931-induced apoptosis. These results suggest that BR931 can increase generation of ROS, leading to DNA damage and p53 phosphorylation, which, in turn, induces the activation of Bax, release of cytochrome-c from mitochondria and activation of caspases, culminating in cell death. PMID- 15135650 TI - Apoptosis-like cell death of human breast cancer cell line MCF-7 induced by buprenorphine hydrochloride. AB - The analgesic buprenorphine hydrochloride (Bph) induced apoptosis-like cell death in the caspase-3-deficient human breast cancer cell line, MCF-7. This apoptosis like cell death activated key molecules in the mitochondrial apoptotic pathway: cytochrome c, caspase-9, caspase-7, and caspase-6. Bph caused the release of fluorescent protein from the mitochondria of MCF-7 cells transfected with the pDsRed2-Mito-vector in a time-dependent manner, suggesting disruption of the mitochondrial membrane. Zn(2+) as high as 2 mM did not inhibit the DNase that took part in this apoptosis. Thus, this unidentified DNase might resemble other DNases involved in apoptosis-like cell death whose activity is not inhibited by zinc ion. PMID- 15135651 TI - Synchronized generation of reactive oxygen species with the cell cycle. AB - A possible appearance of reactive oxygen species (ROS) with the normal cell cycle was studied to find how ROS are generated in cells in relation to the cell cycle. The production of ROS in relation to the cell cycle was examined by determining the changes in intracellular ROS concentrations at different phases of the cell cycle by culturing BALB 3T3 cells in the presence and absence of aphidicolin. The amounts of intracellular ROS and the cell population at specific phases (S and G2/M) were determined as the fluorescence of dichlorodihydrofluorescein and propidium iodide taken up simultaneously by the cells, respectively, by flow cytometry. Although intracellular ROS remained at the control levels when the cell growth was arrested with aphidicolin at the G1 phase, they increased when the arrest was released to result in the increase of the cell population at the S phase. Furthermore, ROS was shown to disturb/stop the cell cycle by means of the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. The cell cycle was regulated through oxidative stress by exposure to hydrogen peroxide and glutathione ethyl ester. The cell cycle was prevented more sensitively in metallothionein-null cells than in the wild type cells. Based on the present observations, we proposed for the first time that ROS are generated synchronously with the normal cell cycle, and that they have to be controlled at certain level for normal progress of the cell cycle. PMID- 15135652 TI - In vitro cytotoxic study of immunoliposomal doxorubicin targeted to human CD34(+) leukemic cells. AB - The expression of CD34 antigen in acute myelogenous leukemias is considered an unfavourable prognosis marker for response to anticancer drugs and duration of remission. This study investigated the applicability of long-circulating immunoliposomes loaded with doxorubicin targeted to CD34 antigen present on MDR(+) human myelogenous leukemia KG-1a cell line. Immunoliposomal doxorubicin showed a higher cytotoxicity against KG-1a cells than non-targeted liposomal doxorubicin, but it did not improve over that of free drug. Although no reversal of doxorubicin resistance was found to occur through its liposomal encapsulation, a therapeutic benefit can be obtained by the selective cytotoxicity observed. Endocytosis studies demonstrated that, after binding to CD34 antigen, the immunoliposomes are not internalized by the KG-1a cells and so the cytotoxic effect might be due to drug released into the space near the cell membrane. Thus, immunotargeting of liposomal doxorubicin to CD34(+) leukemic cells may only provide an ex vivo strategy for site-selective CD34(+) leukemia cell killing. PMID- 15135653 TI - Induction of calcium-activated potassium channel activity by hemin in human erythroleukemia cells. AB - The agent hemin has been demonstrated to be able to initiate a coordinated differentiation program in several cell types. In the present study, we examined the ability of hemin on inducing cell differentiation and Ca(2+)-activated K(+) channel activity in erythroleukemic K562 cells. Treating undifferentiated K562 cells with hemin (0.1 mM) for five days caused these cells to display differentiation-like characteristics including chromatin aggregation, nuclear degradation, pseudopod extension of the membrane and increased hemoglobin production. However, overall cell viability was not significantly changed by the presence of hemin. After hemin treatment for different periods, the Ca(2+) activated K(+) channel was activated by the addition of ionomycin (1 microM), and was inhibited by either clotrimazole, charybdotoxin, or EGTA. Before hemin treatment there was no significant Ca(2+)-activated K(+) channel activity present in undifferentiated K562 cells. After hemin treatment for 5 days, a significant Ca(2+)-activated K(+) channel activity was detected. This increasing Ca(2+) activated K(+) channel activity may be contributed from a subtype of Ca(2+) activated K(+) channel, KCNN4. These results suggest that the ability of hemin to induce increasing Ca(2+)-activated K(+) channel activity may contribute to the mechanism of hemin-induced K562 cell differentiation. PMID- 15135654 TI - Phyllanthus urinaria induces the Fas receptor/ligand expression and ceramide mediated apoptosis in HL-60 cells. AB - Phyllanthus urinaria (P. urinaria), a widely used herb medicine, was tested for the anticancer effect on human myeloid leukemia cells in this study. The water extract of P. urinaria induced the apoptosis of HL-60 cells as demonstrated by morphological change, DNA fragmentation and increased caspase-3 activity. However, normal human peripheral mononuclear cells remained viable under the same treatment. The P. urinaria-induced apoptosis of HL-60 cells was associated with the increased Bax gene expression and decreased Bcl-2 gene expression. In addition, the gene expressions of Fas receptor and Fas ligand, but not p53, were also induced in HL-60 cells dose- and time-dependently. The inhibitor of ceramide synthase, fumonisin B1, completely suppressed the apoptosis induced by P. urinaria and this inhibitory effect of fumonisin B1 could be eliminated by the addition of ceramide. It indicated that the activity of ceramide synthase is critical for the P. urinaria-induced apoptosis in HL-60 cells. The P. urinaria induced apoptosis in HL-60 cells is mediated through a ceramide-related pathway. PMID- 15135655 TI - Effects of naringin on cytosine arabinoside (Ara-C)-induced cytotoxicity and apoptosis in P388 cells. AB - Naringin (NG), a flavonoid in grapefruit and citrus, has been reported to exhibit antioxidant effects and pharmacological actions. Recently, we have reported that NG suppressed the cytotoxicity and apoptosis induced by H(2)O(2), a typical pro oxidant, in mouse leukemia P388 cells. Cytosine arabinoside (1-beta-d arabinofuranosylcytosine; Ara-C) is the most important antimetabolite chemotherapeutic drug used for acute leukemia. It has been suggested that Ara-C induced cytotoxicity is caused by apoptosis, which is mediated by reactive oxygen species (ROS). In this study, we examined the effect of NG on the cytotoxicity and apoptosis in mouse leukemia P388 cells treated with Ara-C. Ara-C caused cytotoxicity in a concentration and time-dependent manner in the cells. N-Acetyl L-cysteine (NAC), cystamine (CysA) or a reduced form of glutathione (GSH), typical antioxidants significantly blocked Ara-C-induced cytotoxicity. Similarly, Ara-C-induced cell death was completely prevented by NG. NG strongly reduced ROS production caused by Ara-C in the cells. NG slightly increased the activities of antioxidant enzymes, catalase and glutathione peroxidase. Ara-C caused apoptosis with nuclear morphological change and DNA fragmentation. NG remarkably attenuated the Ara-C-induced apoptosis. NG completely blocked the DNA damage caused by Ara-C treatment at 6 h using the Comet assay. Our data suggest that NG reduces Ara-C induced oxidative stress through both an inhibition of the generation of ROS production and an increase in antioxidant enzyme activities. Consequently, NG blocked apoptosis caused by Ara-C-induced oxidative stress, resulting in the inhibition of the cytotoxicity of Ara-C. PMID- 15135658 TI - Genetic modification of the heart: exploring necessity and sufficiency in the past 10 years. AB - It has now been approximately a decade since the directed genetic manipulation of genes responsible for normal and abnormal cardiovascular function began. During the past 10 years, thousands of embryonic stem cell lines and hundreds of unique animals carrying defined mutations in genes that impact on cardiac structure and/or function have been made. Homologous recombination in the murine genome has allowed both gene ablation and directed mutations to be placed precisely, while development of powerful transcriptional cassettes that can be exquisitely controlled has allowed both gain and loss of function approaches to be used in order to study a protein's function, determining both necessity and sufficiency. Thus we can now ablate or augment expression of a engineered protein in the heart and are able to effectively remodel the cardiac protein profile and study the consequences of a single genetic manipulation at the molecular, biochemical, cytological and physiologic levels. The ability to effectively manipulate cardiac gene expression via transgenesis or gene targeting enables the investigator to extend correlations to defined mechanisms. This review will revisit these basic technologies, outline the strengths and weaknesses of each, and discuss the advances that are on the experimental horizon. PMID- 15135656 TI - Role of connective tissue growth factor in renal tubular epithelial-myofibroblast transdifferentiation and extracellular matrix accumulation in vitro. AB - Connective tissue growth factor (CTGF) has been reported to play an important role in mediating the profibrotic effects of transforming growth factor-beta (TGF beta) in the pathogenesis of renal fibrosis. To further elucidate the role of CTGF in renal tubular transdifferentiation and extracellular matrix (ECM) metabolism, we examined the time-course of CTGF, alpha-smooth muscle actin (alpha SMA), fibronectin and plasminogen activator inhibitor-1(PAI-1) gene expression upon the stimulation of TGF-beta1 (5 microg/L) in cultured human proximal tubular epithelial cell line (HKC), and further investigated the effects of endogenous CTGF blockade. On reverse transcriptional-polymerase chain reaction (RT-PCR) analysis, TGF-beta1 upregulated CTGF gene expression, preceding that of alpha SMA, fibronectin and PAI-1. The alpha-SMA, fibronectin and PAI-1 mRNA expression induced by TGF-beta1 were significantly inhibited by CTGF antisense oligodeoxynucleotide (ODN) transfection. With prolonged incubation time, CTGF antisense ODN also inhibited intracellular alpha-SMA and PAI-1 protein synthesis, lowered the level of fibronectin and PAI-1 protein secreted into the media, as confirmed by indirect immuno-fluorescence, flow cytometry, enzyme-linked immunosorbent assay (ELISA) and Western blot methods respectively. These results suggested that CTGF may play a crucial role in the renal tubular epithelial transdifferentiation and the following deposition/degradation process of ECM during tubulointerstitial fibrosis. PMID- 15135659 TI - Aquaporin in the heart--only for water? PMID- 15135660 TI - Expression of aquaporin 1 in human cardiac and skeletal muscle. AB - Aquaporins (AQPs) are a family of water channel proteins that assist in maintenance of the cellular osmotic environment and whole body fluid balance. Specialized organ-specific AQPs are important in physiologic and pathologic processes but little is known about AQPs in the human heart. AQP1 has been identified in rodent heart. We investigated the presence and localization of AQP1 in human heart and skeletal muscle using immunohistochemistry and confocal microscopy, western blot and reverse transcriptase-polymerase chain reaction. There was abundant AQP1 present in both cardiac and skeletal muscle. Immunohistochemistry revealed co-localization of AQP1 with vinculin, a t-tubule marker, and caveolin-3. No novel sequences bearing an NPA box motif common to other AQPs were identified in human heart using degenerative PCR analysis. We conclude that AQP1 is present in the human heart. AQP1 co-localizes with t tubular and caveolar proteins. Cardiac AQPs may have a role during osmotic stresses including ischemia/reperfusion and cardiopulmonary bypass. PMID- 15135662 TI - Lysophosphatidylcholine-induced modulation of Ca(2+)-activated K(+)channels contributes to ROS-dependent proliferation of cultured human endothelial cells. AB - Proliferation of endothelial cells plays a crucial role in the process of atherosclerotic plaque destabilization. The major component of oxidized low density lipoprotein lysophosphatidylcholine (LPC) has been shown to promote endothelial proliferation by increasing the production of reactive oxygen species (ROS). Since K(+) channels are known to control the cell cycle, we investigated the role of Ca(2+)-activated K(+) channels (BK(Ca)) in the regulation of LPC induced endothelial proliferation and ROS generation. A significant increase of cell growth induced by LPC (20 micromol/l; cell counts (CCs): +87%, thymidin incorporation: +89%; n = 12, P < 0.01) was observed, which was inhibited by the BK(Ca) inhibitor iberiotoxin (IBX; 100 nmol/l), by the NAD(P)H-oxidase inhibitor diphenyleneiodonium (5 micromol/l) and by transfection with antisense (AS) oligonucleotides against NAD(P)H oxidase, whereas N(G)-monomethyl-l-arginine (l NMMA) further increased LPC-induced cell growth. Using the patch-clamp technique a significant increase of BK(Ca) open-state probability (control: 0.004 +/- 0.002; LPC: 0.104 +/- 0.035; n = 21, P < 0.05) by LPC was observed. Using dichlorofluorescein fluorescence microscopy a significant increase of ROS induced by LPC was reported, that was blocked by IBX and Ca(2+) antagonists. Intracellular Ca(2+) measurements revealed a capacitative Ca(2+) influx caused by LPC. Bioactivity of nitric oxide (NO) was measured using a [(3)H]-cGMP radioimmunoassay. LPC significantly decreased acetylcholine-induced NO synthesis. LPC significantly increased cGMP levels in endothelial cells transfected with AS, which was blocked by IBX. In conclusion, our results demonstrate that LPC activates BK(Ca) thereby increasing ROS production which induces endothelial proliferation. In addition LPC-induced BK(Ca)-activation contributes to increased cGMP levels, if ROS production is prevented by AS. PMID- 15135661 TI - Evolution of expression of cardiac phenotypes over a 4-year period in the beta myosin heavy chain-Q403 transgenic rabbit model of human hypertrophic cardiomyopathy. AB - Hypertrophic cardiomyopathy (HCM), the most common cause of sudden cardiac death in the young, is characterized by a diverse array of cardiac phenotypes evolving over several decades. We have developed transgenic rabbits that fully recapitulate the phenotype of human HCM and provide for the opportunity to delineate the sequence of evolution of cardiac phenotypes, and thus, the pathogenesis of HCM. We determined evolution of biochemical, molecular, histological, structural and functional phenotypes at 4 age-periods in 47 beta myosin heavy chain-glutamine (MyHC-Q)-403 transgenic rabbits. Ca(+2) sensitivity of myofibrillar ATPase activity was reduced very early and in the absence of other discernible phenotypes. Myocyte disarray also occurred early, prior to, and independent of hypertrophy and fibrosis. The latter phenotypes evolved predominantly during puberty in conjunction with activation of stress-related signaling kinases. Myocardial contraction and relaxation velocities were decreased early despite normal global cardiac function and in the absence of histological phenotype. Global cardiac function declined with aging, while left atrial size was increased along with Doppler indices of left ventricular filling pressure. Thus, Ca(+2) sensitivity of myofibrillar ATPase activity is a primary phenotype expressed early and independent of the ensuing phenotypes. Pathogenesis of myocyte disarray, which exhibits age-independent penetrance, differs from those of hypertrophy and fibrosis, which show age-dependent expression. Myocardial dysfunction is an early marker that predicts subsequent development of hypertrophy. These findings in an animal model that recapitulates the phenotype of human HCM, implicate involvement of multiple independent mechanisms in the pathogenesis of cardiac phenotypes in HCM. PMID- 15135663 TI - Cytokines are not upregulated in adriamycin-induced cardiomyopathy and heart failure. AB - Heart failure due to a variety of causes is accompanied by an upregulation of cytokines, such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL 1beta) and interleukin-6 (IL-6). Adriamycin-induced cardiomyopathy (AIC) and heart failure is an important clinical problem. The current study investigated the expression of these cytokines in AIC and heart failure in rats. Both early and late stages of AIC was produced in rats. Myocardial gene expressions for TNF alpha, IL-1beta and IL-6 were examined with DNA microarrays and RT-PCR. Protein levels of these cytokines in both the plasma and the myocardium were also examined by ELISA. In the early stage, myocardial mRNA expression of IL-1beta showed significant increase at 4 and 24 h, peaking at 4 h, while TNF-alpha did not change and IL-6 was undetectable. The protein levels of these three genes did not show any upregulation in the plasma or the heart. In the late stage, heart failure was confirmed by clinical signs as well as homodynamic changes. In this stage, plasma protein levels for TNF-alpha, IL-1beta and IL-6 were not changed. However, myocardial TNF-alpha mRNA expression and protein levels were significantly decreased, while both IL-1beta mRNA and protein levels were not different compared to the control group. IL-6 mRNA expression was undetectable in both normal and adriamycin-treated hearts while its protein level was not changed by adriamycin. Positive control using lipopolysaccharides (LPS) treatment (0.5 mg/kg body weight) for 2 h resulted in a significant increase in these three cytokines in the heart and plasma. These data suggest that an upregulation of cytokines may not be involved in AIC. Heart failure may in fact be accentuated by a downregulation of myocardial TNF-alpha. PMID- 15135664 TI - Ventricular arrhythmias, increased cardiac calmodulin kinase II expression, and altered repolarization kinetics in ANP receptor deficient mice. AB - Cardiac hypertrophy is associated with ventricular arrhythmias and sudden death. The molecular mechanisms that predispose the hypertrophied heart to arrhythmias are not well understood. In mice, deletion of the gene coding for the atrial natriuretic peptide receptor, guanylyl cyclase A (GC-A-/-), causes arterial hypertension, cardiac hypertrophy and sudden death. We used this mouse model to study molecular mechanisms of arrhythmias in the hypertrophied heart. Right and left ventricular monophasic action potential durations (APD) were recorded in isolated, Langendorff-perfused hearts during pacing from the right atrium and ventricle. The atrioventricular (AV) node was ablated to provoke bradycardia. Intracellular Ca(2+) transients were measured in isolated INDO-1 loaded ventricular myocytes. Cardiac expression of Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) was analyzed by western blotting. Polymorphic ventricular arrhythmias (pVT) occurred spontaneously after mechanical AV block in 20/45 hearts from 12-month-old GC-A-/- mice (P < 0.05), but neither in age-matched GC A+/+ hearts nor in hearts from 3-month-old mice of either genotype. Triggered activity preceded pVT. APD were prolonged and systolic Ca(i)(2+) levels were increased in GC-A-/- hearts independently of age. In 12-month-old GC-A-/- hearts only, dispersion of APD and expression levels of CaMKII were increased. CaMKII expression was particularly increased in hearts with pVT. Direct inhibition of CaMKII activation by KN93 (0.5 or 2 microM) or inhibition of Ca(2+)/calmodulin dependent activation of CaMKII by W-7 (25 microM) suppressed pVT in GC-A-/- hearts (P < 0.05) while prolonging APD. The combination of increased CaMKII activity and altered action potential characteristics facilitates ventricular arrhythmias in hypertrophic GC-A-/- hearts. PMID- 15135665 TI - Lidoflazine is a high affinity blocker of the HERG K(+)channel. AB - Lidoflazine is an antianginal calcium channel blocker that carries a significant risk of QT interval prolongation and ventricular arrhythmia. We investigated whether or not lidoflazine inhibits current through the rapid delayed rectifier K(+) channel alpha subunit (encoded by HERG - human ether-a-go-go-related gene), since this channel has been widely linked to drug-induced QT-prolongation. Lidoflazine inhibited potently HERG current (I(HERG)) recorded from HEK 293 cells stably expressing wild-type HERG (IC(50) of approximately 16 nM). It was approximately 13-fold more potent against HERG than was verapamil under similar conditions. On membrane depolarization, I(HERG) inhibition developed gradually, ruling out closed-channel state dependent inhibition. The effect of command voltage on the drug's action suggested that lidoflazine preferentially inhibits activated/open HERG channels. The S6 mutation Y652A largely eliminated the inhibitory action of lidoflazine, whilst the F656A mutation also reduced blocking potency. We conclude: first, that lidoflazine produces high affinity blockade of the alpha subunit of the HERG channel by binding to aromatic amino acid residues within the channel pore and, second, that this is likely to represent the molecular mechanism of QT interval prolongation by this drug. PMID- 15135666 TI - G-CSF treatment increases side population cell infiltration after myocardial infarction in mice. AB - Granulocyte-colony stimulating factor (G-CSF) has been reported to mobilize bone marrow multi-potent stem cells, which differentiate into cardiac myocytes after myocardial infarction (MI). However, there have not been any reports regarding the effect of G-CSF on stem cell infiltration in the MI site. Hearts of mice that had undergone coronary occlusion were isolated and digested with collagenase. Infiltrating cells in the heart were collected using Percoll density gradients. The infiltrating cells were sorted for side population (SP) cells using Hoechst 33342 dye. Hundreds of infiltrating SP cells were found in the heart from 1 to 14 d after MI. There were only a few SP cells in hearts without infarction. Infiltrating SP cells were increased in the 4-d G-CSF treated group compared with the vehicle group (1106 +/- 106 vs. 323 +/- 26/heart, P < 0.05). The infiltration of inflammatory cells was not influenced by the G-CSF treatment. In a separate series of experiments, we confirmed that the infiltrating SP cells were derived from bone marrow. That is, SP cells in the infarcted hearts of mice, which had been transplanted with bone marrow from ROSA 26 (beta-galactosidase transgenic) mice, were positive for beta-galactosidase. In the immunohistochemical examination, Sca-1(+)/CD45(-) cells were existed in the infarcted site after MI. Therefore, SP cells may infiltrate into infarcted heart. G-CSF augmented this kind of stem cell infiltration without increasing inflammatory cells. These results suggest that G-CSF may enhance myocardial regeneration without aggravated inflammation in the infarcted heart. PMID- 15135667 TI - The art of surgery. PMID- 15135668 TI - Relationships among delay of diagnosis, extent of disease, and survival in patients with abdominal carcinoid tumors. AB - BACKGROUND: A correlation between delay in diagnosis of carcinoid and extent of disease and survival was investigated. METHODS: In all, 115 patients with carcinoid were interviewed. Data collected included symptoms, delay of diagnosis, incorrect diagnoses given, extent of disease at diagnosis, and survival. Delay in diagnosis and extent of disease were correlated by regression analysis and the impact on death by Fisher's exact test. RESULTS: Mean delay was 66 months. There was no correlation between delay in diagnosis and extent of disease (R = 0.119, R(2) = 0.014, P = 0.20) or death. However, patients with disease beyond lymph nodes were significantly more likely to die (P = 0.005). CONCLUSIONS: Delays of diagnosis of carcinoid are common but do not impact extent of disease or survival. Extent of disease at diagnosis does impact survival, but is not mitigated by early diagnosis, indicating that outcome is dictated by the biology of the disease. PMID- 15135669 TI - Dacron femoral-popliteal bypass grafts in good-risk claudicant patients. AB - PURPOSE: To evaluate prosthetic femoral-popliteal (FP) grafts as bypass conduits in favorable patients with refractory calf claudication. SETTING: University affiliated urban teaching hospital. METHODS: Cohort study of patency of 6-mm Dacron FP grafts used to treat patients with medically refractory calf claudication meeting the following criteria: nonsmoker; above-knee popliteal artery as an appropriate target vessel; > or =2-vessel tibial outflow; and postoperative anticoagulation and antiplatelet therapy. End points included primary and secondary FP graft patency, need for and outcome of subsequent intervention, limb amputation, and survival. RESULTS: From January 1998 through March 2001, 92 patients underwent 100 Dacron bypass grafts for medically refractory claudication. All underwent serial postoperative graft surveillance using duplex ultrasound. Using the Society for Vascular Surgery/International Society of Cardiovascular Surgery criteria life-table, primary patency was 84%, assisted primary patency was 88%, and secondary patency was 90% at 5 years. Two patients (2%) died during a mean follow-up period of 49 months (range 30 to 68). No limbs required amputation. Among 9 limbs in which Dacron FP bypass failed, 8 underwent subsequent successful autogenous bypass to the popliteal artery. CONCLUSION: In highly selected subjects with favorable features known to increase the likelihood of prosthetic graft patency, prosthetic FP grafts function well. Such patients undergo a simpler, more straightforward procedure than when autogenous FP bypass is performed. Subsequent success of autogenous FP repeat bypass suggests that previous prosthetic FP bypass does not threaten lower extremity arterial outflow. PMID- 15135670 TI - Achieving vascular access success in the quality outcomes era. AB - BACKGROUND: In 1997 the National Kidney Foundation put forth guidelines for hemoaccess through its Dialysis Outcomes Quality Initiative (DOQI). Some centers have been able to meet these standards; most have not. METHODS: A retrospective review was made of our database of more than 3,500 hemoaccess procedures from 1986 to 2003. RESULTS: Our approach, increased use of transposed fistulas and preoperative duplex mapping, has led to a fistula incidence (84%) and prevalence (54%) exceeding DOQI criteria. Meeting the DOQI guideline for thrombectomy rate (0.5 per year) is mostly achieved by increased use of fistulas. Additionally, access monitoring, as well as intraoperative angiography, angioplasty, stenting, and surgical revision, can aid in decreasing the frequency of occlusions (0.45 per year). This operative approach has led to a 98% success rate for surgical thrombectomy, exceeding the DOQI guideline of 85%. Earlier referrals from nephrologists have lowered the catheter use prevalence to 9%. CONCLUSIONS: An aggressive approach to placing fistulas, maintaining the access, and receiving prompt referrals can lead to success in meeting DOQI criteria. PMID- 15135671 TI - Iatrogenic arterial injury is an increasingly important cause of arterial trauma. AB - BACKGROUND: Iatrogenic arterial injuries (IAI) may result from any invasive diagnostic or therapeutic procedure. The relative occurrence and severity of IAI compared with those of penetrating and blunt vascular trauma is unknown. A review of arterial trauma at a university hospital level 1 trauma center, with a focus on iatrogenic injury, forms the basis of this report. METHODS: Patients treated for arterial trauma from January 1994 through October 2002 were identified from prospectively maintained registries. Record review included injury etiology, type of repair, 30-day all-cause mortality, and permanent morbidity. Permanent morbidity was defined as amputation or loss of extremity function. RESULTS: In all, 252 patients required treatment, 85 (33.7%) from IAI, 86 (34.1 %) from penetrating trauma, and 81 (32.1%) from blunt trauma. During the study period, the number of IAIs per year increased. Femoral artery injury from percutaneous intervention (50, 58.8%) was the most frequent IAI; intraoperative injury (including 14 tumor resections and 5 orthopedic procedures) was next most frequent (23, 27.1%). Three patients (3.5%) with IAI had permanent morbidity. The 30-day all-cause mortality was 7.1% (6) for patients with IAI. CONCLUSIONS: Iatrogenic arterial injury is increasingly frequent and caused one third of the arterial trauma at our level 1 trauma center. These data suggest education and training regarding IAI deserves equal priority with the study of penetrating vascular trauma. PMID- 15135673 TI - Outcomes of antireflux surgery in patients with normal preoperative 24-hour pH test results. AB - OBJECTIVE: Preoperative 24-hour pH testing is controversial in surgical patients who have symptomatic gastroesophageal reflux disease (GERD) and endoscopic evidence of esophagitis. The objective of this study was to compare the clinical outcomes of Nissen fundoplication for symptomatic reflux between patients with normal and abnormal preoperative pH testing. METHODS: Patients were selected from a prospective database of patients who underwent laparoscopic esophageal procedures between January 1997 and December 2001 at our institution. Only patients having typical symptoms of GERD (heartburn and/or reflux), preoperative pH testing, manometry, and endoscopy and who had at least 6 months of post operative follow-up were included in the study. Fifteen patients had normal preoperative DeMeester scores (DMS) (median 11.4, range 3.3 to 14.7). These were compared with 208 consecutive patients having abnormal preoperative DMS (median 49.6, range 15.2 to 250). Logistic regression modeling was performed to identify variables significant for poor outcome. Differences between means were tested using appropriate parametric or nonparametric tests. RESULTS: There were no statistically significant differences in demographics, preoperative symptom score (mean 2.9 +/- 1.1 vs. 3.1 +/- 0.68, P = 0.30), or preoperative grade of esophagitis (P = 0.37) between the 2 groups. After a median follow-up of 8.8 months (range 6 to 36), 6 (40%) of the patients having normal preoperative DMS and 17 (8.1%) of the patients having abnormal preoperative DMS continued to have typical GERD symptoms (P <0.01, B error = 0.02). The most significant factor for poor outcome in the regression model was normal preoperative pH (odds ratio 9.02, P <0.01). CONCLUSIONS: Symptomatic GERD patients with normal preoperative 24-hour pH test results have significantly worse subjective outcomes after Nissen fundoplication compared with patients having abnormal preoperative pH test results. To minimize poor symptomatic outcomes after antireflux surgery, a policy of routine preoperative pH testing is advised. PMID- 15135672 TI - Focused high-risk population screening for carotid arterial stenosis after radiation therapy for head and neck cancer. AB - BACKGROUND: Cervical radiation for head and neck cancer has been associated with an increased incidence of carotid arterial stenosis. Modern radiation therapy delivers higher doses with increasing long-term survival. Accordingly, the prevalence of radiation-associated carotid stenosis may be higher than previously reported. Phase I of this prospective study was to establish the prevalence of carotid artery stenosis after high-dose cervical radiation. METHODS: From a prospectively maintained database, we identified patients who had received cervical high-dose radiotherapy (minimum 5,500 cGy). All patients were screened with bilateral carotid arterial duplex ultrasonography. We defined disease as "normal or mild" if the carotid stenosis was <50%, and "significant" if >50%. The relationship between standard demographic risk factors and screening outcomes was then analyzed. RESULTS: Screening was performed in 40 patients (mean age 68.2 years, range 26 to 87). Patients received a mean cumulative radiation dose of 6,420 cGy (range 5,500 to 7,680), with a mean duration of 10.2 years since their last radiation treatment. Sixteen patients (40%) had significant carotid artery stenosis. Patients with and without significant stenosis were comparable in terms of age, radiation dose, tobacco use, comorbidities, and postradiation interval (P = not significant). Six patients (15%) had unilateral complete carotid occlusion and 6 patients (15%) had significant bilateral carotid stenosis. Three patients (7.5%) had sustained a previous stroke after radiation therapy. CONCLUSIONS: The prevalence of carotid arterial disease in patients with prior cervical radiation therapy is clinically significant and warrants aggressive screening as part of routine preradiation and postradiation care. Focused screening of this high-risk population may be cost effective and medically beneficial in terms of risk factor modification and stroke prevention, and will be examined in phase II of this study. PMID- 15135674 TI - Useful benchmarks to evaluate outcomes after esophagectomy and pancreaticoduodenectomy. AB - BACKGROUND: Multiple publications have suggested that outcomes after complex operations are better at high-volume centers. However, of all the potential "outcomes" to measure, only mortality has been studied extensively. The broadest difference in mortality between low- and high-volume centers has been measured after esophagectomy (EG) and pancreaticoduodenectomy (PD). If a low-volume center recorded high mortality, then a broader set of outcomes beyond mortality would be useful for self-assessment. METHODS: Two single-surgeon prospective databases for outcomes of EG and PD were reviewed in a multispecialty clinic within a tertiary referral, resident-training hospital. Between January 1996 and December 2002, 174 consecutive patients underwent EG performed by 1 surgeon (25 cases/y), and 232 consecutive patients underwent PD performed by another surgeon (34 cases/y). We measured hospital and 30-day mortality rate, mean operation time (OR time), mean estimated intraoperative blood loss (EBL), mean length of stay (LOS), and the anastomotic leak rate. These outcomes were compared with those of recently published cases for EG and PD. RESULTS: Mortality for both operations was zero. After EG, OR time was 394 minutes (literature = 336), EBL was 204 mL (literature = 964), transfusion rate was 3.5% (literature = 34%), LOS was 11.1 days (literature = 16.6), leak was 2.9% (literature = 9.1%), and reoperation was 1.7% (literature = not stated). After PD, OR time was 450 minutes (literature = 431), EBL was 382 mL (literature = 1,183), transfusion rate was 7.3% (literature = not stated), LOS was 11.2 days (literature = 17.8), leak was 6.5% (literature = 9.9%), and reoperation was 0.4% (literature = 3.8%). CONCLUSIONS: These 2 single surgeon series provide benchmarks to help better define acceptable outcomes after EG and PD. This assessment demonstrated lower mortality and LOS in a high-volume surgical practice. These outcomes are not associated with OR time but with lower EBL, less need for transfusion, and lower need for reoperation. Anastomotic leaks occurred in both series; however, this was not associated with mortality because of early recognition and the use of nonsurgical minimally invasive techniques. If mortality is high at a low-volume center, then the additional benchmarks of this study, in addition to mortality and LOS, could be used to lower mortality through self-assessment by identifying specific outcomes that need improvement. PMID- 15135675 TI - Is splanchnic perfusion pressure more predictive of outcome than intragastric pressure in neonates with gastroschisis? AB - BACKGROUND: The purpose of this study is to determine whether calculated splanchnic perfusion pressure (SPP) is more predictive of outcome than measured intragastric pressure (IGP) in patients with gastroschisis. METHODS: Retrospective chart review from 1997 through 2003 of 12 patients with gastroschisis. RESULTS: Eight total patients with gastroschisis underwent reduction and had adequate data for analysis. One patient underwent reduction on day of life (DOL) 6; the remainder underwent reduction on DOL 1. All patients had postreduction IGP <20 mm Hg. The correlation coefficient of IGP and date of extubation was 0.20 and of SPP and date of extubation was -0.51. The correlation coefficient of IGP and return of bowel function was -0.06 and of SPP and return of bowel function was -0.50. CONCLUSION: SPP may be more predictive of outcome than IGP after gastroschisis repair. PMID- 15135677 TI - Laparoscopic splenectomy for treatment of splenomegaly. AB - BACKGROUND: Laparoscopic splenectomy presents an advantage over open splenectomy, resulting in shorter hospital stay, decreased blood loss, and fewer operative and postoperative complications. Splenomegaly has long been considered a contraindication for laparoscopic splenectomy; however, in the hands of an experienced surgeon, this technique can be effectively applied to the treatment of splenomegaly. METHODS: Records for patients undergoing laparoscopic splenectomy for splenomegaly between 2000 and 2003, performed by a single surgeon in three community-based hospitals, were reviewed and demographic, operative, and postoperative data compiled. Preoperative diagnoses included B cell lymphoma, hemolytic anemia, and chronic lymphocytic leukemia. RESULTS: Seven patients underwent laparoscopic and hand-assisted laparoscopic splenectomy for splenomegaly during this time period. Splenic weights ranged from 410 to 3,100 g, and average operative time was 86.6 minutes. Estimated blood loss ranged from 50 to 350 ml; average hospital stay was 4.4 days. Two postoperative complications, ie, postoperative bleeding and superficial wound breakdown, were noted. CONCLUSIONS: Laparoscopic and hand-assisted laparoscopic splenectomy are effective methods for treatment of splenomegaly. PMID- 15135676 TI - Outcomes after resection of cholangiocellular carcinoma. AB - BACKGROUND: Cholangiocellular carcinoma (CCC) is a rare primary liver malignancy that arises from intrahepatic bile duct canaliculi and presents as a liver mass. Our purpose is to report operative morbidity and mortality and to determine long term survival after resection for CCC. METHODS: Retrospective review of 31 consecutive patients who underwent resection during a 20-year period. RESULTS: Thirty-day hospital mortality was 3%, and postoperative morbidity was 38%. Kaplan Meier 5-year survival was 35%; mean survival was 37 months; absolute 5-year survival was 33%. Mean survival in stages I, II, IIIA, and IIIC were 57, 33, 26, and 14 months, respectively (P = 0.03 comparing I to >I). Recurrence occurred in 18 patients; 89% were in the liver. Carbohydrate antigen 19-9 >100 U/mL was found to be an indicator of poor prognosis (P = 0.009). CONCLUSIONS: Resection for CCC can be performed with acceptable morbidity and mortality rates and results in good survival and cure. Hepatic recurrence is common. Carbohydrate antigen 19-9 may be useful in determining prognosis. PMID- 15135678 TI - Laparoscopic biliary reconstruction. AB - BACKGROUND: Biliary reconstruction represents a relatively untested frontier in laparoscopy. METHODS: Retrospective review of all patients who underwent laparoscopic biliary operations at Legacy Health System from 1998 to 2003. RESULTS: Seven patients underwent laparoscopic biliary reconstruction. Indications included benign calculous disease in 4 patients, benign stricture on 1 patient, choledochal cyst in 1 patient, and malignant biliary obstruction in 1 patient. Operations performed included choledochoduodenostomy, hepaticojejunostomy, stricturoplasty, choledochal cyst excision with hepaticojejunostomy, and cholecystojejunostomy. Median operative time was 300 minutes. Median hospital stay was 4 days. One perioperative complication of a bowel obstruction required reoperation. Median follow-up was 15 months. One patient died of metastatic cancer 8 months after surgery. All other patients are symptom free with no signs of stricture or recurrent biliary obstruction. CONCLUSIONS: Laparoscopic biliary reconstruction represents a viable treatment option in carefully selected patients. PMID- 15135679 TI - Endorectal ultrasound detection of focal carcinoma within rectal adenomas. AB - BACKGROUND: The misdiagnosis of a rectal adenoma by biopsy and subsequent finding of invasive cancer after transanal excision is associated with a number of pitfalls. Problems include suboptimal therapy for a potentially curable cancerous lesion, potential tumor transgression of the local site with increased chance for local recurrence, and increased potential for more radical surgery or adjuvant chemoradiation. The utility of endorectal ultrasound (ERUS) in guiding treatment decisions of rectal villous adenomas has been reported, but series are small and are from single institutions. To determine the utility of ERUS in the diagnosis of rectal adenomas, we compared diagnosis made by biopsy alone to diagnosis made by a combination of biopsy and ERUS. METHODS: A systematic literature review was performed by way of a PubMed search to find articles with the following terms: "biopsy-negative rectal adenomas," "preoperative ERUS diagnosis," and "surgical histopathology." Five studies met the criteria, thus providing data for 258 adenomas. A quantitative meta-analysis was performed on the data. RESULTS: Among the 258 biopsy-negative rectal adenomas, 24% had focal carcinoma on histopathology. ERUS correctly established a cancer diagnosis in 81% (95% confidence interval 69 to 90) of these misdiagnosed lesions. Thus, ERUS diagnosis of biopsy-negative rectal adenomas could be expected to decrease the need for additional surgery and other associated problems caused by misdiagnosis from 24% to 5%. CONCLUSIONS: ERUS is a useful adjunct to biopsy in the preoperative workup of rectal villous adenomas, and we recommend its routine use. Accurate preoperative assessment allows the surgeon to counsel the patient appropriately regarding the best operation, the perioperative risks, and the chances of local recurrence. PMID- 15135680 TI - Full-thickness intraperitoneal excision by transanal endoscopic microsurgery does not increase short-term complications. AB - PURPOSE: Transanal endoscopic microsurgery (TEM) is a minimally invasive technique for full-thickness excision of benign and malignant rectal neoplasms located 4 to 24 cm above the anal verge. Entrance into the peritoneal cavity during TEM has been regarded as a complication that mandates conversion to open laparotomy for adequate repair of the defect. This study compares the rate of complications arising from TEM with and without intraperitoneal entry. METHODS: Patients undergoing peritoneal entry were compared to those who did not. RESULTS: No perioperative deaths occurred. There was no significant difference in the incidence of postoperative complications. No major complications occurred with peritoneal entry, and all peritoneal entries were closed transanally via endoscope. CONCLUSIONS: Entry into the peritoneum during TEM is not associated with an increased incidence of complication. Entry into the peritoneum during TEM excision does not mandate conversion to open laparotomy but may be safely repaired endoscopically. Lesions likely to be above the peritoneal reflection and within reach of the endoscope (4 to 24 cm) should be considered for TEM excision. PMID- 15135681 TI - Morbid obesity is not a contraindication to kidney transplantation. AB - BACKGROUND: Concern that morbidly obese (body mass index [BMI] 35) kidney transplant recipients have worse outcomes than non-morbidly obese recipients leads many transplant centers to deny them the benefit of kidney transplantation. These concerns are supported by guidelines recently published by the American Society of Transplantation. However, successfully transplanted morbidly obese persons have a survival advantage over those that remain on dialysis. It is our practice to evaluate morbidly obese transplant candidates for transplantation under the same criteria used for nonobese candidates. This report reviews our experience with morbidly obese kidney transplant recipients having a three year follow-up. METHODS: Outcomes for 23 morbidly obese (BMI 35, range 37 to 56) kidney transplant recipients transplanted between January 1995 and February 2000 were compared with 224 nonobese (BMI 25) kidney recipients transplanted during the same period. RESULTS: Patient and graft survivals were similar between both groups. Although 3-year graft survival for morbidly obese recipients of cadaver organs was 75% compared with 90% for the nonobese group, this finding was not statistically significant, and 3-year graft survival was 100% for morbidly obese recipients of living donor organs compared with 91% for nonobese recipients. Morbidly obese recipients had significantly longer hospital stays, higher readmission rates, and a higher wound infection rate than nonobese recipients. CONCLUSIONS: We found that morbidly obese persons have 3-year graft and patient survivals similar to nonobese persons. Although they also have greater complications and greater numbers of days in the hospital, we believe these reasons are not sufficient to deny morbidly obese persons the survival and quality-of-life advantages of kidney transplantation. PMID- 15135682 TI - Criteria for establishing the adequacy of a sentinel lymphadenectomy. AB - BACKGROUND: Criteria are needed that could be used to terminate a sentinel lymphadenectomy for breast cancer prior to removing every sentinel lymph node, without increasing false negative rates. METHODS: Quantitative information on the radioactivity and color of sentinel lymph nodes removed from 541 breast cancer patients was correlated with pathologic information to determine when a sentinel lymphadenectomy could be terminated based on characteristics of the initially removed nodes. RESULTS: Tumor was found in the first two sentinel lymph nodes removed in 127 of 129 node-positive patients. In 65% of patients who were able to be evaluated, the most radioactive lymph node was a positive lymph node. When any axillary lymph node was blue, then the first tumor-containing sentinel lymph node was also blue. CONCLUSIONS: Removal of the most radioactive lymph node does not insure accurate assessment of the axilla. Removal of two sentinel lymph nodes accurately staged 98.4% of node-positive patients and 99.6% of the entire study population. PMID- 15135683 TI - Secondary operations are frequently required to complete the surgical phase of therapy in the era of breast conservation and sentinel lymph node biopsy. AB - BACKGROUND: Breast conservation therapy (BCT) is an oncologically equivalent and cosmetically preferable alternative to mastectomy for most early-stage breast cancers. The number of operations required to complete the surgical phase of therapy with BCT has not been widely reported. METHODS: From our institutional tumor registry, we reviewed the records of all patients receiving primary surgical therapy for breast cancer from January 1, 1998, to June 30, 2002. There were 204 patients with 210 breast cancers in the cohort. These cancers were initially managed with either BCT (n = 150) or mastectomy (modified radical mastectomy or total mastectomy with sentinel lymph node biopsy) (n = 60). We compared the percentages of patients in each group who required additional surgeries to obtain clear margins, manage axillary disease, or otherwise complete the surgical phase of therapy. Patients with secondary surgery related to long term local recurrence were excluded. RESULTS: Fifty-one percent of patients initially managed with BCT required additional surgery compared with 12% in the mastectomy group (P <0.05). Factors independently associated with multiple surgeries among all patients included management with BCT (odds ratio [OR] 5.4, P = 0.01) and positive margins at initial excision (OR 4.7, P <0.01). Significant independent predictors of positive margins included BCT (OR 11.9, P <0.01); disease stage (OR 6.7, P <0.01); submission of supplemental margins in addition to the main specimen (OR 2.8, P = 0.03); and positive nodes (OR 1.1, P = 0.04). Breast conservation was ultimately successful in 95% of patients who underwent BCT. CONCLUSIONS: Patients undergoing BCT may require multiple surgeries to reconcile successful breast conservation with sound oncologic resection. PMID- 15135684 TI - Bilateral reduction mammoplasty in combination with lumpectomy for treatment of breast cancer in patients with macromastia. AB - BACKGROUND: Bilateral reduction mammoplasty has been described as a surgical option for treatment of breast cancer in women with large, pendulous breasts. Using this technique can provide unique surgical oncologic challenges. METHODS: Retrospective chart review. RESULTS: Thirty-seven patients were identified. The average weight of tissue removed was 653 g. Tumor size ranged from 0.6 to 5.2 cm. One patient had microscopically positive surgical margins on final pathology. The tumor bed was not marked, and completion mastectomy was required. Two patients had unexpected malignancy in the contralateral breast. One patient required completion mastectomy for positive nonoriented margins. In another patient, tissue from the contralateral side was oriented, and mastectomy was avoided. CONCLUSIONS: Bilateral reduction mammoplasty with lumpectomy is an ideal option in women with macromastia. We recommend marking the tumor bed on the flaps and pedicle as well as orienting tissue removed from the contralateral side as maneuvers that will decrease need for completion mastectomy. PMID- 15135685 TI - Combat trauma experience with the United States Army 102nd Forward Surgical Team in Afghanistan. AB - BACKGROUND: The United States Army 102nd Forward Surgical Team (FST) was deployed to Kandahar Airfield, Afghanistan, from August 2002 to March 2003, in support of Operation Enduring Freedom. The unit's primary mission was to provide trauma surgical support to units of the 101st and 82nd Airborne Divisions, to coalition special operations units, and to allied Afghan militia forces. The FST's mission was expanded to include humanitarian assistance. METHODS: The mission was accomplished in the austere environment of Kandahar Airfield, Afghanistan. The FST was set up in a corner of the abandoned Kandahar International Airport terminal. The team's supporting facility was a 44-bed combat support hospital at Bagram Airbase near Kabul. Patients arrived by ground ambulance, local transportation, and MediVac helicopter. Evacuation of casualties, when necessary, was by fixed-wing aircraft. Patient data were retrospectively reviewed. RESULTS: The team performed 112 surgeries on 90 patients during the course of 7 months. Three patients were female (all children). Twenty patients were <19 years old. Trauma accounted for 78% of cases; the remainders were nontrauma or elective cases. Sixty-seven percent of these surgeries were performed on Afghan militia and civilians, 30% on United States soldiers, and 3% on other coalition forces. Mechanism of injury included gunshot wounds (34%), blasts (18%), motor vehicle crashes (14%), stab wounds (5%), and other trauma (7%). By physiological system, the trauma cases were broken down into extremity (44%), head and neck (17%), multisystem (13%), trunk (8%), and vascular (3%). CONCLUSIONS: "Damage control" operations necessitating multiple trips to the operating room were the norm. Hypothermia from blood loss was often exacerbated by exposure before evacuation and prolonged transport in helicopters. This was aggressively treated with passive, conductive, and active rewarming techniques. Stabilization and evacuation to higher echelons of care was common. PMID- 15135686 TI - Roux-en-Y divided gastric bypass results in the same weight loss as duodenal switch for morbid obesity. AB - BACKGROUND: The 2 weight loss procedures most commonly performed in the United States are Roux-en-Y gastric bypass (RYGBP) and lateral gastrectomy with duodenal switch (BPD/DS). RYGB is a restrictive procedure, whereas BPD/DS relies on mild restriction of intake as well as malabsorption. Many physicians believe that weight loss is greater after BPD/DS than after RYGBP. However, these procedures have not been compared using groups of patients operated on by the same surgeons at the same institution. METHODS: We compared weight loss (expressed as percent of excess body weight [%EBW]) after 1 and 2 years in patients who underwent open RYGB or BPD/DS at our institution. RESULTS: Average length of stay was longer in BPD/DS patients than in those undergoing RYGBP (8.7 vs. 5.9 days, P <0.05). Anastomotic leaks were higher after BPD/DS (6% vs. 3%), but the difference did not achieve statistical significance. Mortality did not differ between the 2 groups (0.8% vs. 0.9%). In the group of patients followed-up for 1 to 2 years, age and distribution of men and women did not differ. Those patients undergoing BPD/DS had higher body mass index (59 vs. 55, P <0.05). Weight loss expressed as %EBW was similar between the 2 groups: 54% versus 53% at 1 year and 67% versus 64% at 2 years. CONCLUSIONS: Our data suggested that weight loss expressed as %EBW is similar between patients undergoing RYGBP and those undergoing BPD/DS. However, BPD/DS was associated with a longer hospital stay. PMID- 15135687 TI - The hand-held ultrasound examination for penetrating abdominal trauma. AB - BACKGROUND: An evaluation of hand-held ultrasonography (US) in the assessment of penetrating torso trauma has not yet been reported. METHODS: A 2.4 kg hand-held ultrasound device was used to examine penetrating trauma victims in an exam designated as the Hand-Held Focused Assessment with Sonography for Trauma (HHFAST). Results were compared with other US examinations including formal FAST (FFAST), computed tomography, diagnostic peritoneal lavage, operative and autopsy findings, and serial examination. Performance considered both the detection of fluid and injuries requiring intervention. RESULTS: The HHFAST was excellent for detecting free intraperitoneal fluid, which had 100% specificity for peritoneal penetration, but was only moderately sensitive for injuries requiring therapy. CONCLUSIONS: Hand-held sonography can quickly detect intraperitoneal fluid, which has good test performance in determining the presence of an intra-abdominal injury. Negative FAST examinations after penetrating trauma should be followed up with another diagnostic modality. PMID- 15135688 TI - A case-match analysis of failed prior bariatric procedures converted to resectional gastric bypass. AB - BACKGROUND: Failure to lose weight or intractable symptoms after bariatric surgery presents a complex diagnostic and management challenge. The outcome of a standardized surgical approach to this problem has not been well described. Conversion of failed bariatric procedures to a resectional gastric bypass (RGB) can achieve symptomatic relief and acceptable weight loss. METHODS: We reviewed all patients in a prospectively maintained database who underwent reoperative bariatric surgery over a 4-year period. Reoperative patients (RO) were case matched (by age, body mass index, and comorbidities) in a 1:2 ratio with control patients undergoing an initial bariatric procedure (IN). RESULTS: Twenty-seven reoperative patients and 54 case-matched control patients were identified. Mean body mass index was 42 in the RO group versus 45 in the IN group (P = not significant). Indications for conversion were weight gain (89%), dysphagia/emesis (30%), esophagitis (19%), and marginal ulcer (7%). All patients in both groups underwent RGB (subtotal gastrectomy with Roux-Y gastrojejunostomy). Compared with IN patients, the RO patients had significantly longer operative times (420 versus 268 minutes), greater blood loss (650 versus 315 cc), longer time to oral intake (3.1 versus 2.2 days), and longer hospital stays (6.5 versus 4.7 days), all P <0.01. There were no deaths or anastomotic leaks in either group. Excess body weight lost at 6 months was 46% for RO versus 54% for IN (P = 0.02). One-year excess weight lost was 71% for RO versus 77% for IN (P = not significant). All RO patients achieved symptomatic relief, and no patient required further bariatric revision. There was significant improvement in weight-related comorbidity in each group. CONCLUSIONS: Conversion of failed bariatric procedures to RGB, although technically demanding, resulted in relief of presenting symptoms, significant 6 month and 1-year weight loss, and improvement of major comorbidities. Conversion of failed bariatric procedures to resectional gastric bypass can achieve results comparable with those of patients undergoing an initial bariatric procedure. PMID- 15135689 TI - Methylenetetrahydrofolate reductase gene C677T and A1298C polymorphisms, plasma homocysteine, folate, and vitamin B12 levels and the extent of coronary artery disease. AB - The question of whether mild hyperhomocysteinemia is a risk factor for coronary artery disease (CAD) has long been debated and is still unclear. We investigated whether there is a link between methylenetetrahydrofolate reductase (MTHFR) gene C677T and A1298C polymorphisms or plasma homocysteine and CAD. This is a case control study that included 2,121 consecutive patients (cases) with angiographically proved CAD and 617 patients without CAD (controls). MTHFR gene C677T and A1298C polymorphisms, plasma homocysteine, folate, and vitamin B(12) concentrations were determined and coronary angiography was performed in all subjects. The distribution of MTHFR gene C677T genotypes in patients (or controls) was: CC-genotype in 915 cases, 43.1% (266 controls, 43.1%); CT-genotype in 955 cases, 45.0%, (283 controls, 45.9%); and TT-genotype in 251 cases, 11.9% (68 controls, 11.0%) (p = 0.84). The distribution of MTHFR gene A1298C genotypes in patients (or controls) was: AA-genotype in 973 cases, 45.9% (281 controls, 45.5%); AC-genotype in 905 cases, 42.7% (284 controls, 46.0%); and CC-genotype in 243 cases, 11.4% (52 controls, 8.5%) (p = 0.07). Patients with CAD had higher levels of plasma homocysteine (12.9 +/- 5.1 vs 11.9 +/- 4.5 micromol/L, p <0.001) and lower levels of folate (9.5 +/- 3.1 vs 9.9 +/- 3.8 ng/ml, p = 0.008) than controls. After adjustment for other risk factors for CAD, plasma homocysteine (p = 0.89), MTHFR gene C677T (p = 0.38), or A1298C polymorphisms (p = 0.13) were not independent correlates of CAD. This study demonstrated that MTHFR gene C677T or A1298C polymorphisms are not associated with the presence of angiographic CAD. Although there is an apparent association between elevated levels of homocysteine and CAD, this association is not independent of conventional cardiovascular risk factors. PMID- 15135690 TI - Usefulness of myocardial contrast echocardiography in predicting collateral blood flow in the presence of a persistently occluded acute myocardial infarction related coronary artery. AB - Adequate collateral blood flow at rest can sustain myocardial viability despite persistent occlusion of the infarct-related artery (IRA) in acute myocardial infarction (AMI). This has therapeutic and prognostic implications. Studies addressing the value of intravenous myocardial contrast echocardiography (MCE) to detect collateral blood flow after AMI in humans are limited. Accordingly, 70 consecutive patients with AMI underwent low-power intravenous MCE using a Sonovue infusion 7 to 10 days after thrombolysis. Myocardial perfusion detected by MCE was analyzed (qualitatively and quantitatively) in the akinetic segments in 20 patients (29%) with an occluded IRA who subsequently underwent revascularization. Contractile reserve, which is a marker of myocardial viability, was assessed with low-dose dobutamine 12 weeks after mechanical revascularization. Of the 102 akinetic segments (32%), 37 (36%) showed contractile reserve. Contractile reserve was present in 24 of the 29 segments (83%) with homogenous contrast opacification and absent in 60 of the 73 segments (82%) with reduced/absent opacification. Quantitative peak contrast intensity, microbubble velocity, and myocardial blood flow were significantly higher (p <0.0001) in the segments with contractile reserve than in those without contractile reserve. Multiple logistic regression analysis using electrocardiographic, biochemical, and myocardial contrast echocardiographic markers of collateral blood flow showed that MCE (odds ratio 26.0, 95% confidence interval 6.3 to 108.0, p <0.001) was the only independent predictor of collateral blood flow as demonstrated by the presence of contractile reserve. MCE may thus be used as a reliable bedside technique for the accurate evaluation of collateral blood flow in the presence of an occluded IRA after AMI. PMID- 15135691 TI - Quantitative myocardial contrast echocardiography for prediction of thrombolysis in myocardial infarction flow in acute myocardial infarction. AB - Clinical evaluation of arterial patency in acute ST-elevation myocardial infarction (STEMI) is unreliable. We sought to identify infarction and predict infarct-related artery patency measured by the Thrombolysis In Myocardial Infarction (TIMI) score with qualitative and quantitative intravenous myocardial contrast echocardiography (MCE). Thirty-four patients with suspected STEMI underwent MCE before emergency angiography and planned angioplasty. MCE was performed with harmonic imaging and variable triggering intervals during intravenous administration of Optison. Myocardial perfusion was quantified offline, fitting an exponential function to contrast intensity at various pulsing intervals. Plateau myocardial contrast intensity (A), rate of rise (beta), and myocardial flow (Q = A x beta) were assessed in 6 segments. Qualitative assessment of perfusion defects was sensitive for the diagnosis of infarction (sensitivity 93%) and did not differ between anterior and inferior infarctions. However, qualitative assessment had only moderate specificity (50%), and perfusion defects were unrelated to TIMI flow. In patients with STEMI, quantitatively derived myocardial blood flow Q (A x beta) was significantly lower in territories subtended by an artery with impaired (TIMI 0 to 2) flow than those territories supplied by a reperfused artery with TIMI 3 flow (10.2 +/- 9.1 vs 44.3 +/- 50.4, p = 0.03). Quantitative flow was also lower in segments with impaired flow in the subtending artery compared with "normal" patients with TIMI 3 flow (42.8 +/- 36.6, p = 0.006) and all segments with TIMI 3 flow (35.3 +/- 32.9, p = 0.018). An receiver-operator characteristic curve derived cut-off Q value of <11.3, representing impaired myocardial flow, was 73% sensitive and 67% specific for TIMI <3 flow at angiography. Thus, qualitative MCE identifies patients with STEMI but provides no information regarding infarct-related artery patency, whereas quantitative MCE can predict impaired flow in patients with acute STEMI. PMID- 15135692 TI - Care concordant with guidelines predicts decreased long-term mortality in patients with unstable angina pectoris and non-ST-elevation myocardial infarction. AB - Data are sparse regarding the long-term benefit of care concordant with clinical practice guidelines in patients presenting with unstable angina pectoris and non ST-segment elevation myocardial infarction (UAP/NSTEMI), particularly in the general care setting. We extended follow-up in a preexisting cohort of 275 patients hospitalized with primary UAP/NSTEMI. Using Cox models, we compared long term mortality between patients who received care concordant with > or =80% of 8 important guideline recommendations during the index hospitalization and patients who did not. Among all study patients, 68% received guideline-concordant care. During follow-up (median 9.4 years) 49% of patients died. Patients with UAP/NSTEMI who received guideline-concordant care had significantly decreased long-term mortality compared with those who received guideline-discordant care (hazards ratio [HR] 0.45, 95% confidence interval [CI] 0.32 to 0.64). Guideline concordant care remained associated with decreased mortality after adjusting for other predictors of long-term mortality (HR 0.57, 95% CI 0.39 to 0.84) and after adjustment for the propensity to receive guideline-concordant care (HR 0.61, 95% CI 0.43 to 0.88). The benefit of guideline-concordant care relative to discordant care was preserved in high-risk populations shown to be less likely to receive guideline-concordant care, including patients with advanced age, congestive heart failure, elevated serum creatinine, and prior myocardial infarction. Care concordant with UAP/NSTEMI clinical practice guidelines is associated with substantially improved long-term survival. Our findings endorse the approach adopted by authors of clinical practice guidelines in generalizing evidence-based medicine to usual clinical care. In firmly establishing the benefit of consensus guidelines, the foundation is set for efforts to improve practitioner compliance with these standards. PMID- 15135693 TI - Use of a scoring model combining clinical, exercise test, and echocardiographic data to predict mortality in patients with known or suspected coronary artery disease. AB - The aim of this study was to derive and validate a model for predicting mortality by combining clinical, exercise testing, and echocardiographic data in patients with known or suspected coronary artery disease. We studied 5,679 patients (aged 62 +/- 12 years; 3,231 men) who were followed for a mean of 3 years after treadmill exercise echocardiography. Patients were randomly divided into 2 groups of equal size. (1) The modeling group underwent multivariate analysis to define independent predictors of mortality. Three hundred bootstrap resamplings were performed to determine parameter coefficients. Patients were divided into 5 risk categories according to their composite score and survival rate in each category was estimated by the Kaplan-Meier method. (2) The validation group comprised patients for whom the risk model was applied. Patients were divided into 5 risk categories based on data obtained from the modeling group. During follow-up, 315 patients died (151 in the modeling group). Independent predictors of mortality were exercise wall motion score index (chi-square 22.4, p <0.0001), workload (chi square 17.1, p <0.0001), male gender (chi-square 15.4, p <0.0001), and age (chi square 5.5, RR 1.02, 95% confidence interval 1 to 1.04; p = 0.02). Application of the composite score in the validation group resulted in an effective stratification of patients for mortality and cardiac events. This study provides a model for assessing risk of death by combining clinical, exercise testing, and echocardiographic data using a single composite score. PMID- 15135694 TI - Impact of body mass index on in-hospital outcomes following percutaneous coronary intervention (report from the New York State Angioplasty Registry). AB - Although obesity traditionally has been considered a risk factor for coronary revascularization, recent data from registry studies have shown a possible protective effect of obesity on outcomes after percutaneous coronary intervention (PCI). Using data from the New York State Angioplasty database over a 4-year period, we analyzed 95,435 consecutive patients who underwent PCI. Classification of body mass index (BMI) was: underweight (<18.5 kg/m(2)), healthy weight (18.5 to 24.9 kg/m(2)), overweight (25 to 29.9 kg/m(2)), moderate obesity (class I) (30 to 34.9 kg/m(2)), severe obesity (class II) (35 to 39.9 kg/m(2)), and very severe obesity (class III) (>40 kg/m(2)). In-hospital postprocedural mortality and complications were compared among these groups. Compared with healthy weight patients, patient with class I or II obesity had lower in-hospital mortality and major adverse cardiac events (MACE) (combined death, myocardial infarction, and emergency surgery), whereas patients at the extremes of BMI (underweight and class III obese patients) had significantly higher mortality and MACE rates. Adjusted hazards ratios for in-hospital mortality according to BMI were: underweight (2.69), healthy weight (1.0), overweight (0.90), class I obese (0.74), class II obese (0.67), and class III obese (1.63). Patients at the extremes of BMI (<18.5 and >40 kg/m(2)) were at increased risk of MACEs, including mortality after PCI, whereas patients who were moderately to severely obese (BMIs 30 to 40 kg/m(2)) were at lower risk than healthy weight patients. PMID- 15135695 TI - Randomized comparison of balloon angioplasty versus silicon carbon-coated stent implantation for de novo lesions in small coronary arteries. AB - The objectives of this study were to compare the short- and long-term clinical and angiographic outcomes of conventional balloon angioplasty versus elective silicon carbide-coated stent implantation in de novo lesions in small coronary arteries (<3.0 mm). Angioplasty of small coronary arteries is associated with high restenosis rates. The beneficial effects of stent implantation on restenosis rates and clinical outcomes have been suggested by observational studies. However, randomized trials comparing balloon angioplasty with stenting show conflicting results. A total of 496 patients with a de novo stenosis in a coronary artery, with a diameter between 2.0 and 3.0 mm, were randomly assigned to balloon angioplasty (n = 246) or stent implantation (n = 250). One-year clinical and 6-month angiographic follow-up were performed. Baseline characteristics were similar for the 2 groups, with 16% having diabetes and 77% complex lesions (type B or C). Crossover to stents occurred in 29% of patients randomized to balloon angioplasty. At 6 months, angiographic restenosis rates were 25% versus 21% for the angioplasty and stent groups, respectively (p = NS). One-year clinical follow-up showed no difference in occurrence of major adverse cardiac events. Elective silicon carbon-coated stent implantation in small coronary arteries is safe and effective, with no beneficial effect on angiographic or clinical outcome compared with balloon angioplasty with provisional stenting. PMID- 15135696 TI - C-reactive protein levels among women of various ethnic groups living in the United States (from the Women's Health Study). AB - The distribution of C-reactive protein (CRP) levels was compared among 24,455 white, 475 black, 357 Asian, and 254 Hispanic women, all of whom are participants in the Women's Health Study. Median CRP levels were significantly higher among black women (2.96 mg/L, interquartile range [IQR] 1.19 to 5.86) than among their white (2.02 mg/L, IQR 0.81 to 4.37), Hispanic (2.06 mg/L, IQR 0.88 to 4.88), and Asian (1.12 mg/L, IQR 0.48 to 2.25) counterparts. As expected, women taking hormone replacement therapy had higher baseline CRP levels than women not taking hormone replacement therapy. No differences in low-density lipoprotein cholesterol or total cholesterol levels were observed between ethnic groups. In multivariate regression models, body mass index was a significant (p <0.001) predictor of elevated CRP concentrations among all race/ethnic groups, and control for body mass index substantially attenuated the differences noted in CRP levels across race/ethnic groups, particularly among black women. Control for all measured modifiable risk factors for cardiovascular disease did not entirely explain CRP differences. Among these women, the distribution of CRP levels varied significantly between the various race/ethnic groups. PMID- 15135697 TI - Systolic and diastolic regional myocardial motion of pacing-induced versus idiopathic left bundle branch block with and without left ventricular dysfunction. AB - To demonstrate systolic and diastolic asynchrony in patients with left bundle branch block (LBBB), tissue Doppler imaging (TDI) of 4 different walls was performed in 27 normal controls, 29 patients with right ventricular pacing and normal left ventricular (LV) ejection fraction (EF; pacing LBBB), and 35 patients with idiopathic LBBB. Patients with idiopathic LBBB were further classified into those with LVEF >50% and those with LVEF <35%. Asynchrony was calculated as the coefficient of variation of the time intervals from the QRS complex to the peak systolic velocity and to the peak of early diastolic relaxation. Patients with pacing and idiopathic LBBB had significantly longer QRS durations (162 +/- 20 vs 92 +/- 7 ms, p <0.001) and larger systolic (15.9 +/- 5.0% vs 4.1 +/- 2.1%, p <0.001) and diastolic (3.7 +/- 2.0% vs 1.4 +/- 0.6%, p <0.001) asynchrony than controls. Those with idiopathic LBBB and low EF had significantly larger diastolic asynchrony (5.7 +/- 2.1%) than those with pacing LBBB (2.9 +/- 1.1%) and those with idiopathic LBBB and normal EF (2.0 +/- 0.6%). Diastolic asynchrony was the only independent factor that correlated with LVEF (r = -0.64, p <0.001). Thus, idiopathic LBBB with LV dysfunction is characterized not only by systolic but also by diastolic asynchrony. PMID- 15135698 TI - Relation between achieved heart rate and outcomes in patients with atrial fibrillation (from the Atrial Fibrillation Follow-up Investigation of Rhythm Management [AFFIRM] Study). AB - Many patients with atrial fibrillation (AF) are treated with rate control and anticoagulation. However, the relation between the degree of heart rate (HR) control and clinical outcome is uncertain. We assessed whether lower achieved HR at rest and/or lower achieved exercise HR was associated with improved prognosis, quality of life (QoL), and functional status among patients in the AFFIRM study. Patients in the rate control arm and who were in AF at baseline and 2 months were included. Patients were grouped by quartile of achieved HR at rest (44 to 69, 70 to 78, 79 to 87, 88 to 148 beats/min) and achieved exercise HR following a 6 minute walk (53 to 82, 83 to 92, 93 to 106, 107 to 220 beats/min). QoL measurements and functional status were also analyzed. Complete data were available for 680 patients for achieved HR at rest, 349 patients for achieved exercise HR, and 118 patients for QoL. Survival free from cardiac hospitalization and overall survival were not significantly different among quartiles of achieved HR at rest (p = 0.19 and p = 0.8, respectively) or achieved exercise HR (p = 0.77 and p = 0.14, respectively). After controlling for covariates, there remained no significant relation between either achieved HR at rest or achieved exercise HR and event-free survival (hazard ratio 0.95, p = 0.35 and hazard ratio 0.98, p = 0.81) or overall survival (hazard ratio 1.03, p = 0.70 and hazard ratio 1.22, p = 0.13). Furthermore, there was no significant association between achieved HR and QoL measurements, New York Heart Association functional class, or 6-minute walking distance. After 2 months of drug titration, neither achieved HR at rest nor achieved exercise HR predicted survival free from cardiovascular hospitalization, overall survival, QoL, or functional status among patients with AF. PMID- 15135699 TI - Relation of unrecognized hypervolemia in chronic heart failure to clinical status, hemodynamics, and patient outcomes. AB - Clinically unrecognized intravascular volume overload may contribute to worsening symptoms and disease progression in patients with chronic heart failure (CHF). The present study was undertaken to prospectively compare measured blood volume status (determined by radiolabeled albumin technique) with clinical and hemodynamic characteristics and patient outcomes in 43 nonedematous ambulatory patients with CHF. Blood volume analysis demonstrated that 2 subjects (5%) were hypovolemic (mean deviation from normal values -20 +/- 6%), 13 subjects (30%) were normovolemic (mean deviation from normal values -1 +/- 1%), and 28 subjects (65%) were hypervolemic (mean deviation from normal values +30 +/- 3%). Physical findings of congestion were infrequent and not associated with blood volume status. Increased blood volume was associated with increased pulmonary capillary wedge pressure (p = 0.01) and greatly increased risk of death or urgent cardiac transplantation during a median follow-up of 719 days (1-year event rate 39% vs 0%, p <0.01 by log-rank test). Systolic blood pressure was significantly lower in hypervolemic patients than in those with normovolemia or hypovolemia (107 +/- 2 vs 119 +/- 2 mm Hg, p = 0.008), and hypotension was independently associated with increased risk of hypervolemia in multivariate analysis (odds ratio 2.64 for a 10 mm Hg decrease in systolic blood pressure, 95% confidence interval 1.13 to 6.19, p = 0.025). These findings demonstrate that clinically unrecognized hypervolemia is frequently present in nonedematous patients with CHF and is associated with increased cardiac filling pressures and worse patient outcomes. PMID- 15135700 TI - Role of adenosine monophosphate deaminase-1 gene polymorphism in patients with congestive heart failure (influence on tumor necrosis factor-alpha level and outcome). AB - The Cytosin-->thymidin transition at codon 12 of the adenosine monophosphate deaminase-1 (AMPD1) gene results in a complete loss of its catalytic activity. The increased conversion of adenosine monophosphate to adenosine, which in turn attenuates the expression of tumor necrosis factor-alpha (TNF-alpha) expression, has been suggested as a putative mechanism for prolonged survival in patients with congestive heart failure (CHF) carrying the mutant AMPD1 allele. Therefore, the impact of this polymorphism on circulatory TNF-alpha concentrations and outcome in patients with CHF should be studied. The AMPD1 genotype of each patient with CHF (n = 90; idiopathic dilated cardiomyopathy n = 53; coronary artery disease n = 20; other n = 17) was determined by direct sequencing. Serum TNF-alpha concentrations were measured by enzyme-linked immunosorbent assay. We found 66 patients (75.6%) to be homozygous for the wild-type allele (AMPD1 +/+), and 20 patients (22.2%) were heterozygous and 2 were homozygous (2.2%) for the mutant AMPD1 allele (AMPD1 +/- or -/-). TNF-alpha serum concentrations were 4.2 +/- 2.0 pg/ml for the AMPD1 +/+ genotype and 5.3 +/- 2.9 pg/ml for the AMPD1 +/- and -/- genotypes (p = 0.045). A downregulation of TNF-alpha in patients carrying the mutant allele could therefore be not detected. However, Kaplan-Meier analysis demonstrated a significantly prolonged survival without heart transplantation or revival from sudden death in the AMPD1 +/- & -/- group (p = 0.020). Multivariate analysis identified the AMPD1 wild-type genotype as an independent risk factor (odds ratio 9.34, 95% confidence interval 1.78 to 48.96). The mutant AMPD1 allele, in the context of CHF, is associated with a prognostic benefit. The underlying mechanism of TNF-alpha is unrelated. PMID- 15135701 TI - Usefulness of ultrasound lung comets as a nonradiologic sign of extravascular lung water. AB - The "comet-tail" is an ultrasound sign detectable with ultrasound chest instruments; this sign consists of multiple comet-tails fanning out from the lung surface. They originate from water-thickened interlobular septa and would be ideal for nonradiologic bedside assessment of extravascular lung water. To assess the feasibility and value of ultrasonic comet signs, we studied 121 consecutive hospitalized patients (43 women and 78 men; aged 67 +/- 12 years) admitted to our combined cardiology-pneumology department (including cardiac intensive care unit); the study was conducted with commercially available echocardiographic systems including a portable unit. Transducer frequencies (range 2.5 to 3.5 MHz) were used. In each patient, the right and left chest was scanned by examining predefined locations in multiple intercostal spaces. Examiners blinded to clinical diagnoses noted the presence and numbers of lung comets at each examining site. A patient lung comet score was obtained by summing the number of comets in each of the scanning spaces. Within a few minutes, patients underwent chest x-ray, with specific assessment of extravascular lung water score by 2 pneumologist-radiologists blinded to clinical and echo findings. The chest ultrasound scan was obtained in all patients (feasibility 100%). The imaging time per examination was always <3 minutes. There was a linear correlation between echocardiographic comet score and radiologic lung water score (r = 0.78, p <0.01). Intrapatient variations (n = 15) showed an even stronger correlation between changes in echocardiographic lung comet and radiologic lung water scores (r = 0.89; p <0.01). In 121 consecutive hospitalized patients, we found a linear correlation between echocardiographic comet scores and radiologic extravascular lung water scores. Thus, the comet-tail is a simple, non-time-consuming, and reasonably accurate chest ultrasound sign of extravascular lung water that can be obtained at bedside (also with portable echocardiographic equipment) and is not restricted by cardiac acoustic window limitations. PMID- 15135702 TI - Incidence of thrombotic stent occlusion during the first three months after sirolimus-eluting stent implantation in 500 consecutive patients. AB - Sirolimus-eluting stents have been used in our institution for all percutaneous interventions, without clinical or anatomic exclusion criteria, as part of the Rapamycin-Eluting Stent Evaluated At Rotterdam Cardiology Hospital registry. We analyzed the incidence of (sub)acute stent thrombosis after sirolimus-eluting stent implantation in an unselected population of 510 consecutive patients. At 3 month follow-up, (sub)acute stent thrombosis was diagnosed in 2 patients (0.4%) 6 hours and 11 days after the procedure, respectively. These cases occurred in diabetic women with complex coronary lesions. Intravascular ultrasound examination showed inadequate stent expansion and uncovered distal dissection as possible mechanical explanations for the thrombosis. PMID- 15135703 TI - Usefulness of myocardial viability or ischemia in predicting long-term survival for patients with severe left ventricular dysfunction undergoing revascularization. AB - In 107 patients with coronary disease and severe left ventricular dysfunction, we examined the prognostic power of viability identified by dobutamine stress echocardiography. At a mean follow-up of 27 months, patients with viable myocardium who underwent revascularization had a significant survival advantage over all other patients (2-year survival 83.5% vs 57.2%, p = 0.0037). PMID- 15135704 TI - Comparative pharmacodynamic evaluation of eptifibatide and tirofiban HCl in patients undergoing percutaneous coronary intervention (the TAM1 Study). AB - We performed a comparative analysis of platelet aggregation inhibition achieved with the glycoprotein IIb/IIIa inhibitors eptifibatide and tirofiban HCl in patients who underwent percutaneous coronary intervention and used light transmission aggregometric assays with D-phenylalanyl-L-prolyl-L-arginine chloromethyl ketone as an anticoagulant and 20 micromol of adenosine diphosphate as an agonist. Taking into account the differences in clinical efficacy of these 2 drugs in large trials investigating percutaneous coronary intervention, we hypothesized that the variable clinical effects might be related to variability in the magnitude and consistency of platelet aggregation inhibition achieved with dosing regimens of these glycoprotein IIb/IIIa inhibitors. PMID- 15135705 TI - Comparison of treatment outcomes in patients > or =80 years undergoing transradial versus transfemoral coronary intervention. AB - We assessed the effect of transradial access (vs transfemoral access) for percutaneous coronary intervention on postprocedure length of stay and patient outcomes (in-hospital complications and all-cause and cardiac death at 6 and 12 months) in 225 elderly patients (> or =80 years old). Raw differences between transradial and transfemoral accesses were compared, and 3 forms of propensity score analysis were used to determine the true effect of transradial access. After matching to adjust for baseline differences in patient characteristics, remaining differences in outcomes and postprocedure length of stay were small and not statistically significant at the 95% level, but a decrease in postprocedural length of stay of nearly 1 day was observed and likely was not due to chance. Transradial access in patients > or =80 years old undergoing percutaneous coronary intervention should be preferred due to equivalent success rate and safety and likely reduction in postprocedural hospitalization. PMID- 15135706 TI - Gender and place of death from coronary artery disease in Washington State. AB - Men may be more likely than women to die outside the hospital from coronary artery disease, whereas women may be as likely as men to die inside the hospital. This study used Washington State death records to compare death rates from coronary artery disease inside and outside the hospital for women and men from 1980 to 2001. For in-hospital deaths, the men:women ratio of deaths was 1.6 in 1980 and 2001; for out-of-hospital deaths, the ratio decreased slightly from 2.6 in 1980 to 2.3 in 2001. These findings support the contention that, despite significant decreases in mortality, men continue to have higher rates of death outside and inside the hospital than do women, although there has been a slight decrease in male:female mortality for deaths outside the hospital. PMID- 15135707 TI - Educational attainment and myocardial infarct-related congestive heart failure (the Determinants of Myocardial Infarction Onset Study). AB - We assessed educational attainment and complications during hospitalization in 3,882 patients with acute myocardial infarction for the Determinants of Myocardial Infarction Onset Study. Educational attainment had a graded, inverse association with prevalence of congestive heart failure (CHF) during hospitalization, with the lowest prevalence among patients who attended college (odds ratio 0.75; 95% confidence interval, 0.57 to 0.99; p for trend 0.02), with similar findings among men, women, whites, and blacks. PMID- 15135708 TI - Effects of daily stress on autonomic cardiac control in patients with coronary artery disease. AB - Emotional stress has been implicated in the development and progression of coronary artery disease, with 1 proposed causal pathway being changes in cardiac autonomic tone. One hundred thirty-five patients with coronary artery disease underwent 48 hours of ambulatory electrocardiographic monitoring and completed activity and mood diaries every 20 minutes while awake. Random-effects model analyses associated higher levels of negative emotions (e.g., anger, stress, sadness) with decreases in high- and low-frequency power, whereas higher levels of positive emotion were related to an increase in low-frequency power, independent of age, posture, and medications. PMID- 15135709 TI - Coronary artery disease and opioid use. AB - Over the past 20 years, we have observed a paucity of morbidity and mortality due to cardiovascular disease among drug users in a methadone maintenance clinic. The present study investigated whether long-term exposure to opiates or opioids is associated with decreased severity of coronary artery disease (CAD) by comparing 98 decedents with methadone or opiates (M/O) in their blood at autopsy with 97 frequency-matched decedents without M/O. Severe CAD was found significantly less often in M/O-positive decedents (5 of 98) than in M/O-negative decedents (16 of 97). Multiple logistic regression analysis contrasting those with moderate or severe CAD to those with no or mild CAD yielded an odds ratio of 0.43 (95% confidence interval 0.20 to 0.94) for M/O positivity after adjustment for potential confounding. Long-term opiate exposure thus may mitigate CAD severity and its often fatal consequences. PMID- 15135710 TI - Anatomical observations of the pulmonary veins with intracardiac echocardiography and hemodynamic consequences of narrowing of pulmonary vein ostial diameters after radiofrequency catheter ablation of atrial fibrillation. AB - Intracardiac echocardiography was used to explore pulmonary venous (PV) anatomy and to monitor PV stenosis in 31 patients referred for radiofrequency catheter ablation at PV ostia. Interindividual variations in PV anatomy and insertion in the left atrium were observed. Narrowing of PV ostia after radiofrequency catheter ablation did not produce significant hemodynamic changes. PMID- 15135711 TI - Acute and long-term outcome of transvenous cryoablation of midseptal and parahissian accessory pathways in patients at high risk of atrioventricular block during radiofrequency ablation. AB - The ability of transvenous cryothermal catheter ablation to create reversible lesions (cryomapping) and to avoid catheter dislodgment (cryoadherence) has been shown to be safe and highly effective in elimination of atrioventricular nodal reentrant tachycardia. In addition, cryoablation may be useful in the management of perinodal accessory pathways, but its efficacy and safety in patients at high risk of atrioventricular block during radiofrequency catheter ablation is unknown. This study prospectively evaluated the efficacy and safety of cryoablation in patients with midseptal and parahissian accessory pathways. PMID- 15135712 TI - Inconsistent electrocardiographic testing for syncope in United States emergency departments. AB - The electrocardiogram has diagnostic and prognostic value in the evaluation of syncope, and consensus guidelines suggest routine electrocardiographic testing. An analysis of a nationally representative survey suggests that electrocardiographic testing is performed inconsistently in patients presenting with syncope to United States emergency departments, even in high-risk patients, such as the elderly and hospitalized. Variation in electrocardiographic testing represents an opportunity to improve the care of patients presenting with syncope to emergency departments. PMID- 15135713 TI - Effects of levosimendan on circulating pro-inflammatory cytokines and soluble apoptosis mediators in patients with decompensated advanced heart failure. AB - This randomized, placebo-controlled trial showed that levosimendan administration causes a significant reduction of circulating proinflammatory cytokine interleukin-6 and soluble apoptosis mediators, such as soluble Fas and Fas ligand in patients with decompensated heart failure. These immunomodulatory effects may lead to improvement of symptoms and echocardiographic markers of cardiac contractile performance in these patients. PMID- 15135714 TI - Usefulness of the third heart sound in predicting an elevated level of B-type natriuretic peptide. AB - Third heart sounds were sought in 100 consecutive outpatients who had B-type natriuretic peptide (BNP) levels measured within 8 hours. Mean BNP levels were significantly higher in those with a third heart sound. The presence of a third heart sound was 41% sensitive and 97% specific for elevated BNP levels. PMID- 15135715 TI - Carcinoid heart disease associated with primary ovarian carcinoid tumor. AB - Because the venous drainage of the ovary bypasses the portal circulation, carcinoid heart disease in patients with primary ovarian carcinoid tumors may develop in the absence of liver metastasis. We describe 4 patients who presented with symptomatic carcinoid heart disease in association with primary ovarian carcinoid tumor. PMID- 15135716 TI - Trends in the use of diagnostic imaging in patients hospitalized with acute pulmonary embolism. AB - From 1979 to 2001, the proportion of imaging tests by computed tomography (CT), ventilation perfusion (VQ) lung scan, pulmonary angiography, and venous ultrasound was assessed in patients with pulmonary embolism (PE) from the National Hospital Discharge Survey. By 2001, there was a higher proportion of imaging tests with CT than VQ scans (36% vs 32%). Even so, in the United States, a large proportion of patients continued to have VQ scans. PMID- 15135717 TI - Validation of the i-STAT handheld activated clotting time for use with bivalirudin. AB - Bivalirudin is being used more frequently as an anticoagulant in the cardiac catheterization laboratory. Newer devices, used to measure activated clotting time (ACT), have not been thoroughly tested for use with bivalirudin. One such device, the i-STAT ACT, measures the generation of activated thrombin to determine the level of anticoagulation. Our study demonstrated a high level of agreement between the i-STAT ACT and the Hemochron ACT in patients anticoagulated with bivalirudin. In addition, the i-STAT was shown to have an extremely high degree of reproducibility. PMID- 15135718 TI - Effectiveness of sotalol treatment in symptomatic Brugada syndrome. AB - We describe a 53-year-old man with recurrent syncopal events and a malignant family history who was treated for 13 years with sotalol drug therapy with no further occurrence of Brugada syndrome symptoms. Genetic testing revealed that he carried a Brugada syndrome sodium channel SCN5A mutation (4189delT). This finding suggests that sotalol may be of therapeutic benefit in such patients. PMID- 15135720 TI - Famotidine and long QT syndrome. AB - Numerous drugs have been implicated in causing a prolonged QT interval and Torsades de pointes. However, the association of famotidine and acquired long QT syndrome has rarely been reported. We report 2 cases of famotidine-associated acquired long QT syndrome. PMID- 15135719 TI - Spontaneous coronary artery dissection associated with sexual intercourse. AB - We report a case of spontaneous coronary artery dissection associated with sexual intercourse in a 32-year-old, morbidly obese patient. This is the first report of spontaneous coronary artery dissection induced by coital activity and the sixth such report in a male patient. PMID- 15135721 TI - Proceedings of the editorial board meeting of The American Journal of Cardiology on 8 March 2004. PMID- 15135722 TI - Twelve eponymous signs of aortic regurgitation, one of which was named after a patient instead of a physician. PMID- 15135723 TI - Is depression following acute myocardial infarction an independent risk for mortality? PMID- 15135724 TI - Metabolic syndrome, inflammation, and exercise. PMID- 15135725 TI - The dark side of light: the damaging effects of UV rays and the protective efforts of MAP kinase signaling in the epidermis. PMID- 15135726 TI - AP endonuclease and poly(ADP-ribose) polymerase-1 interact with the same base excision repair intermediate. AB - Base excision repair (BER) is a defense system that protects cells from deleterious effects secondary to modified or missing DNA bases. BER is known to involve apurinic/apyrimidinic endonuclease (APE) and DNA polymerase ss (ss-pol) among other enzymes, and recent studies have suggested that poly(ADP-ribose) polymerase-1 (PARP-1) also plays a role by virtue of its binding to BER intermediates. The main role of APE is cleavage of the DNA backbone at abasic sites, and the enzyme also can catalyze 3'- to 5'-exonuclease activity at the cleaved abasic site. Photocross-linking studies with mouse embryonic fibroblast (MEF) cell extracts described here indicated that APE and PARP-1 interact with the same APE-cleaved abasic site BER intermediate. The model BER intermediate used includes a synthetic abasic site sugar, i.e. tetrahydrofuran (THF), in place of the natural deoxyribose. APE cross-linked efficiently with this intermediate, but not with a molecule lacking the 5'-THF phosphate group, and the same property was demonstrated for PARP-1. The addition of purified APE to the MEF extract reduced the amount of PARP-1 cross-linked to the BER intermediate, suggesting that APE can compete with PARP-1. APE and PARP-1 were antagonists of each other in in vitro BER related reactions on this model BER intermediate. These results suggest that PARP-1 and APE can interact with the same BER intermediate and that competition between these two proteins may influence their respective BER related functions. PMID- 15135727 TI - The role of TDP1 from budding yeast in the repair of DNA damage. AB - The TDP1 gene encodes a protein that can hydrolyze certain types of 3'-terminal phosphodiesters, but the relevance of these catalytic activities to gene function has not been previously tested. In this work we engineered a point mutation in TDP1 and present evidence that, as per design, it severely diminishes tyrosyl-DNA phosphodiesterase enzyme activity without affecting protein folding. The phenotypes of yeast strains that express this mutant show that the contribution of TDP1 to the repair of two kinds of damaged termini-induced, respectively, by camptothecin (CPT) and by bleomycin-strongly depends on enzyme activity. In routine assays of cell survival and growth the contribution of this activity is often overshadowed by other repair pathways. However, the value of TDP1 in the economy of the cell is highlighted by our discovery of several phenotypes that are evident even without deliberate inactivation of parallel pathways. These non redundant mutant phenotypes include increased spontaneous mutation rate, transient accumulation of cells in a mid-anaphase checkpoint after exposure to camptothecin and, in cells that overexpress topoisomerase I (Top1), decreased survival of camptothecin-induced damage. The relationship between the role of TDP1 in Saccharomyces and its role in metazoans is discussed. PMID- 15135728 TI - Disruption of Drosophila Rad50 causes pupal lethality, the accumulation of DNA double-strand breaks and the induction of apoptosis in third instar larvae. AB - The Rad50/Mre11/Nbs1 protein complex has a crucial role in DNA metabolism, in particular in double-strand break (DSB) repair through homologous recombination (HR). To elucidate the role of the Rad50 protein complex in DSB repair in a multicellular eukaryote, we generated a Rad50 deficient Drosophila strain by P element mediated mutagenesis. Disruption of Rad50 causes retarded development and pupal lethality. To investigate the mechanism of pupal death, brains and wing imaginal discs from third instar larvae were studied in more detail. Wing imaginal discs from Rad50 mutant larvae displayed a 3.5-fold increase in the induction of spontaneous apoptotic cells in comparison to their heterozygous siblings. This finding correlates with increased levels of phosphorylated histone H2Av, indicating an accumulation of DSBs in Rad50 mutant larvae. A 45-fold increase in the frequency of anaphase bridges was detected in the brains of Rad50 deficient larvae, consistent with a role for Rad50 in telomere maintenance and/or replication of DNA. The induction of DSBs and defects in chromosome segregation are in agreement with a role of Drosophila Rad50 in repairing the DSBs that arise during replication. PMID- 15135729 TI - Role of nucleotide- and base-excision repair in genotoxin-induced neuronal cell death. AB - Base-excision (BER) and nucleotide-excision (NER) repair play pivotal roles in protecting the genomes of dividing cells from damage by endogenous and exogenous agents (i.e. environmental genotoxins). However, their role in protecting the genome of post-mitotic neuronal cells from genotoxin-induced damage is less clear. The present study examines the role of the BER enzyme 3-alkyladenine DNA glycosylase (AAG) and the NER protein xeroderma pigmentosum group A (XPA) in protecting cerebellar neurons and astrocytes from chloroacetaldehyde (CAA) or the alkylating agent 3-methyllexitropsin (Me-Lex), which produce ethenobases or 3 methyladenine (3-MeA), respectively. Neuronal and astrocyte cell cultures prepared from the cerebellum of wild type (C57BL/6) mice or Aag(-/-) or Xpa(-/-) mice were treated with 0.1-50 microM CAA for 24h to 7 days and examined for cell viability, DNA fragmentation (TUNEL labeling), nuclear changes, and glutathione levels. Aag(-/-) neurons were more sensitive to the acute (>20 microM) and long term (>5 microM) effects of CAA than comparably treated wild type neurons and this sensitivity correlated with the extent of DNA fragmentation and nuclear changes. Aag(-/-) neurons were also sensitive to Me-Lex at comparable concentrations of CAA. In contrast, Xpa(-/-) neurons were more sensitive than either wild type or Aag(-/-) neurons to CAA (>10 microM), but less sensitive than Aag(-/-) neurons to Me-Lex. Astrocytes from the cerebellum of wild type, Aag(-/-) or Xpa(-/-) mice were essentially insensitive to CAA at the concentrations tested. These studies demonstrate that BER and NER are required to protect neurons from genotoxin-induced cell death. PMID- 15135730 TI - The Werner syndrome protein confers resistance to the DNA lesions N3 methyladenine and O6-methylguanine: implications for WRN function. AB - The Werner syndrome (WS) protein (WRN), a DNA helicase/exonuclease, is required for genomic stability and avoidance of cancer. Current evidence suggests that WRN is involved in the resolution of stalled and/or collapsed replication forks. This function is indicated, in part, by replication defects in WS cells and by hypersensitivity to agents causing major structural aberrations in DNA that block replication. We show here that antisense suppression of WRN in two human glioma cell lines reproduces hallmarks of the drug cytotoxicity profile of WS cells, namely, hypersensitivity to 4-nitroquinoline 1-oxide, camptothecin and hydroxyurea. We also show that antisense-treated cells are hypersensitive to methyl-lexitropsin, a site-specific alkylating agent that produces mainly N3 methyladenine, a cytotoxic and replication-blocking lesion. Antisense-treated cells are hypersensitive to O(6)-methylguanine adducts as well, but only when repair by O(6)-methylguanine-DNA methyltransferase is lacking. Our results illustrate the drug sensitivity caused by deficiency of WRN in a uniform genetic background. They extend the WRN DNA damage sensitivity spectrum to methyl base adducts that can result in blocked replication, and suggest that WRN may be required for resumption of processive replication when incomplete repair of DNA damage leaves blocking lesions at forks. The evidence that highly disparate lesions fall within the purview of WRN, and that abrogating DNA repair can reveal dependence on WRN, suggests that WRN may protect the genome from the lethal, mutagenic and carcinogenic effects of widely diverse DNA damage arising from endogenous processes and environmental agents. PMID- 15135731 TI - Functional interactions between the MutL and Vsr proteins of Escherichia coli are dependent on the N-terminus of Vsr. AB - The crystal structure of the Escherichia coli Vsr endonuclease bound to a C(T/G)AGG substrate revealed that the DNA is held by a pincer composed of a trio of aromatic residues which intercalate into the major groove, and an N-terminus alpha helix which lies across the minor groove. We have constructed an N-terminus truncation (Delta14) which removes most of the alpha helix. The mutant is still fairly proficient in mediating very short patch repair. However, its endonuclease activity is considerably reduced and, in contrast to that of the wild type protein, cannot be stimulated by MutL. We had shown previously that excess Vsr in vivo causes mutagenesis, probably by inhibiting the participation of MutL in mismatch repair. The Delta14 mutant has diminished mutagenicity. In contrast, four enzymatically inactive mutants, with intact N-termini, are as mutagenic as the wild type protein. On the basis of these results we suggest that MutL causes a conformational change in the N-terminus of Vsr which enhances Vsr activity, and that this functional interaction between Vsr and MutL decreases the ability of MutL to carry out mismatch repair. PMID- 15135732 TI - XPC lymphoblastoid cells defective in the hMutSalpha DNA mismatch repair complex exhibit normal sensitivity to UVC radiation and normal transcription-coupled excision repair of DNA cyclobutane pyrimidine dimers. AB - Nucleotide excision (NER) is generally considered to comprise two partially distinct subpathways. Global genomic repair (GGR) removes damage from the genome overall and transcription-coupled repair (TCR) selectively excises damage from transcribed DNA. Cells from individuals belonging to xeroderma pigmentosum (XP) complementation group C are defective in GGR but retain a functional TCR pathway. DNA mismatch repair (MMR) corrects replication errors but can also process DNA damage. It has been suggested that the essential hMutSalpha and hMutLalpha MMR protein complexes are also required for effective excision of UV-induced cyclobutane pyrimidine dimers (CPD) by TCR. We have combined an MMR and an XPC defect in a human lymphoblastoid cell line. The MMR-defective XPC cells were defective in the hMutSalpha mismatch recognition complex that comprises hMSH2 and hMSH6. They were not detectably more sensitive to killing by UV than their MMR proficient counterparts and were able to excise CPDs from an actively transcribed DNA strand. We conclude efficient TCR does not depend on a functional hMutSalpha complex. PMID- 15135733 TI - On the role of proofreading exonuclease in bypass of a 1,2 d(GpG) cisplatin adduct by the herpes simplex virus-1 DNA polymerase. AB - UL30, the herpes simplex virus type-1 DNA polymerase, stalls at the base preceding a cisplatin crosslinked 1,2 d(GpG) dinucleotide and engages in a futile cycle of incorporation and excision by virtue of its 3'-5' exonuclease. Therefore, we examined the translesion synthesis (TLS) potential of an exonuclease-deficient UL30 (UL30D368A). We found that UL30D368A did not perform complete translesion synthesis but incorporated one nucleotide opposite the first base of the adduct. This addition was affected by the propensity of the enzyme to dissociate from the damaged template. Consequently, addition of the polymerase processivity factor, UL42, increased nucleotide incorporation opposite the lesion. The addition of Mn(2+), which was previously shown to support translesion synthesis by wild-type UL30, also enabled limited bypass of the adduct by UL30D368A. We show that the primer terminus opposite the crosslinked d(GpG) dinucleotide and at least three bases downstream of the lesion is unpaired and not extended by the enzyme. These data indicate that the primer terminus opposite the lesion may be sequestered into the exonuclease site of the enzyme. Consequently, elimination of exonuclease activity alone, without disrupting binding, is insufficient to permit bypass of a bulky lesion by this enzyme. PMID- 15135734 TI - Early studies on recombination and DNA repair in Ustilago maydis. AB - This historical review covers the period 1960 to mid-1980s. The first experiments were carried out at the John Innes Institute, Bayfordbury, Hertford, with a one year interlude in the Department of Genetics, University of Washington, Seattle. In 1965, I moved to the National Institute for Medical Research, Mill Hill, London, and became head of a new Genetics Division. The research on Ustilago was divided broadly into (1) experimental genetic studies, and (2) DNA enzymology, largely under the direction of the late Geoffrey Banks. The approaches involved isolating and characterizing mutants defective in repair and recombination (the first in any eukaryotic organism), with the longer term aim of identifying the function of genes through studies of enzymes and proteins which interact with DNA. An enzyme capable of recognizing mismatched bases in DNA was identified. A novel method exploited the inducible nitrate reductase gene, and revealed relationships between recombination, mutation, repair, transcription and cell survival. Several different studies provide strong evidence for the presence of an inducible repair pathway, dependent on recombination. Much more recently, the revolution in molecular genetics has been in exploited in several laboratories working with Ustilago maydis, and these have produced some completely new insights into recombination and repair. PMID- 15135735 TI - The potential of immunostimulatory CpG DNA for inducing immunity against genital herpes: opportunities and challenges. AB - Herpes simplex virus type 2 (HSV-2) invades human genital tract mucosa and following local replications can be rapidly transmitted via peripheral nerve axons to the sacral ganglia where it can establish latency. Reactivation of the latent viral reservoir results in recurrent ulcers in the genital region. Innate immunity, the first line of defence during both primary and recurrent genital herpes infections, is crucial during the period of acute infection to limit early virus replication and to facilitate the development of an appropriate specific acquired immunity. Recent developments in immunology reveal that the mammalian innate immune systems use Toll-like receptor (TLR) to specifically sense evolutionary conserved molecules such as bacterial DNA in pathogens. Recently, local-vaginal delivery of CpG containing oligodeoxynucleotide (ODN), a synthetic mimic of bacterial DNA, holds substantial promise as a strong inducer of innate immunity against genital herpes infections in the animal models of the disease. These preclinical observations provide a scientific ground work for introduction of this novel intervention strategy to clinic. This review aims to highlight recent developments and future challenges in use of immunostimulatory CpG ODN for inducing immunity against genital herpes infection and disease. PMID- 15135736 TI - The antiviral effect of interferon-beta against SARS-coronavirus is not mediated by MxA protein. AB - Severe acute respiratory syndrome (SARS) is caused by a novel coronavirus termed SARS-CoV. No antiviral treatment has been established so far. Interferons are cytokines which induce the synthesis of several antivirally active proteins in the cell. In this study, we demonstrated that multiplication of SARS-CoV in cell culture can be strongly inhibited by pretreatment with interferon-beta. Interferon-alpha and interferon-gamma, by contrast, were less effective. The human MxA protein is one of the most prominent proteins induced by interferon beta. Nevertheless, no interference with SARS-CoV replication was observed in Vero cells stably expressing MxA. Therefore, other interferon-induced proteins must be responsible for the strong inhibitory effect of interferon-beta against SARS-CoV. PMID- 15135737 TI - A one step quantitative RT-PCR for detection of SARS coronavirus with an internal control for PCR inhibitors. AB - In this study, we report a one step quantitative RT-PCR assay for severe acute respiratory syndrome (SARS) diagnosis. The overall detection rate for clinical samples collected from Days 1 to 9 of disease onset is 86.2% and the specificity of the assay is 100%. To detect false negative results due to PCR inhibitors or faulty RNA extraction, we have further modified this one step RT-PCR assay for simultaneous detection of severe acute respiratory syndrome (SARS) coronavirus (CoV) and 18S ribosomal RNA (rRNA) as an internal positive control. PMID- 15135738 TI - Virological exploration of individuals with discordant HIV screening tests. AB - Screening for HIV infection can use many algorithms. When two different HIV antibody assays are used, discordant results may occur. To discriminate between HIV seroconversion, HIV variant infection and false positive reactivity, 30 consecutive subjects with two discordant HIV antibody-screening assays were extensively investigated for HIV infection. No subject had HIV seroconversion or reached HIV seropositivity criteria after a follow-up of 3 months. By contrast 36% became HIV negative by the use of both HIV screening assays. p24 Antigen, HIV 1 RNA, HIV-1 DNA, HIV-2 DNA assays and HIV isolation by sensitive culture were unable to identify HIV infection in the 30 subjects with discordant HIV screening assays. The data suggest that the use of two HIV screening assays increase false positive HIV results without increasing clinical sensitivity. To compliment follow-up of HIV screening, early testing for HIV RNA could be useful to identify or eliminate a recent infection. PMID- 15135739 TI - Characterization of human immunodeficiency virus type-1 from HIV-1 seropositive cases with undetectable viremia. AB - BACKGROUND: Human immunodeficiency virus type 1 (HIV-1) viral load has become a standard of care among HIV-1-infected patients; however, a small number of patients have undetectable viral load even though they have never been treated. METHODS: By using RT-PCR and DNA-PCR, and followed by sequencing and phylogenetic analyses, a detailed molecular characterization was carried out from five HIV-1 seropositive patients who had undetectable viral load by commercially available ultrasensitive viral load assays. RESULTS: Of the four patients whose plasmas were available, viral RNAs were detected in three of them by using an in-house RT PCR in at least one of the three regions (integrase, protease or envgp41). The fourth patient had positive RT-PCR signals in these regions only when RNA isolated from the supernatant of cocultivated patient PBLs with PHA-stimulated HIV-1 negative donor PBLs was used. Further analysis of DNA extracted from the PBMCs revealed that four of the five patients had detectable proviral sequences in at least two of the three regions. The fifth patient had only positive PCR results in all three regions when DNA isolated from PHA-stimulated patient's PBLs was used. Phylogenetic analysis of protease and envgp41 regions revealed that three patients were infected with subtype B viruses while the remaining two patients were infected with subtype C and CRF02_AG viruses. These subtypes coincided with geographic origin and known molecular epidemiology of HIV-1 infection. CONCLUSION: These data provide evidence that both subtype B and non-B HIV-1 infection can result in undetectable viral load in HIV-1-infected patients and that efforts should continue to further characterize these viruses. PMID- 15135740 TI - Performance of the OraQuick and Hema-Strip rapid HIV antibody detection assays by non-laboratorians. AB - Rapid HIV antibody tests (RT) now permit HIV screening in settings where laboratory personnel may not be available. This study assessed the ability of 99 individuals with no laboratory experience to conduct two RT, OraQuick and Hema Strip; these results were compared with those generated by laboratory professionals. All participants received written instructions and one-half also received a short demonstration. Error rates ranged from 2.1% to 4.6% with or without a demonstration. However, the number of invalid tests was greatly reduced when participants received a demonstration. Appropriate RT training for non laboratorians and continued monitoring of HIV RT performance in non-laboratory settings is recommended. PMID- 15135741 TI - Evaluation of rubella IgM enzyme immunoassays. AB - BACKGROUND: Rubella virus generally causes a mild fever, rash illness similar in clinical presentation to infections by other viruses including measles and parvovirus B19. Rubella infections in pregnant women in the first trimester carry a high risk of congenital rubella syndrome (CRS) which can result in severe congenital defects in the infants. The goal of rubella immunization programs is therefore to eliminate CRS. The primary test for the laboratory confirmation of rubella is IgM serology. It is therefore important to evaluate currently available commercial rubella IgM immunoassays to ensure high quality rubella diagnostic testing. STUDY DESIGN: In this study, we compared the performance of seven commercial rubella IgM enzyme immunoassays (EIA) (Meddens, Denka Seiken, Behring, Wampole, Captia, Sigma and Abbott Axsym) using well-defined panels of sera from rubella and non-rubella/rash-illness cases. RESULTS: The Meddens, Denka Seiken, Behring and Wampole rubella IgM EIAs all performed similarly for sensitivity (range of 74.1-76.8%) and specificity (range of 93.9-96.1%). Relative to the other assays, the Axsym had a higher sensitivity (78.9%) but lower specificity (86.5%). The Captia assay had the lowest overall sensitivity (66.4%), while the Sigma assay had a lower specificity (85.6%) in relation to the other assays. CONCLUSIONS: In conclusion, the Meddens, Denka Seiken, Behring and Wampole rubella IgM EIAs are comparable in their overall performance with respect to sensitivity and specificity. PMID- 15135742 TI - Prognostic value of proliferative responses to HIV-1 antigen in chronically HIV infected patients under antiretroviral therapy. AB - BACKGROUND: In the chronic stage of HIV infection T cell proliferative responses to HIV antigens are rare, mostly of low level, and the influence of responses on antiretroviral therapy is not known. OBJECTIVES: To determine a potential correlation between HIV-specific proliferative responses and the subsequent course of infection under antiretroviral therapy. STUDY DESIGN: Proliferation assays were performed with freshly isolated blood mononuclear cells from 45 chronically HIV-infected HAART treated individuals using HIV-p24, other recall antigens, and mitogens as stimulants. Virus load was monitored at the time of stimulation and during 33 months follow-up. RESULTS: A proliferative response to HIV antigen stimulation was detectable in 7 of 45 patients (15.5% responders). This group showed elevated reactions against tetanus toxoid and tuberculin, whereas reactions against standard mitogens were equal in the HIV responder and nonresponder groups. None of the seven HIV-specific responders had a blood virus load rebound of more than 1000 genome copies/ml during follow-up, whereas in 50% of the non-responders higher virus rebounds occurred. CD4 cell levels were slightly higher in the responder group, but mostly independent of virus rebound within the non-responders. Only four patients with high and continuous virus rebound experienced a significant CD4 cell decline. CONCLUSIONS: In patients under HAART, HIV-specific proliferative response is frequently related to anamnestic antigen responses and an enduring control of virus replication. PMID- 15135743 TI - National epidemic of Lordsdale Norovirus in the UK. AB - BACKGROUND: In early 2002 reports of outbreaks of gastroenteritis reached unprecedented levels in the UK. Forty five Norovirus outbreaks were reported in January 2002. OBJECTIVES: The objective of the study was to determine whether the outbreaks were Noroviral in origin and if so whether they represented a homogeneous or heterogeneous collection of Noroviruses by applying EIA and sequence analysis to representative faecal samples. STUDY DESIGN: Faecal specimens were collected during the week of highest incidence from 21 outbreaks in a variety of health care settings including hospitals and nursing homes. The outbreaks occurred in geographically distinct regions of the UK and samples were collected by reference laboratories in Glasgow, Manchester, Bristol and Southampton. RESULTS: The samples were all positive for Noroviruses by negative stain electron microscopy (EM) and Lordsdale virus (LV) EIA, therefore reverse transcriptase polymerase chain reaction (RT-PCR) amplification and nucleotide sequencing of the Norovirus RNA polymerase gene was performed on amplicons from samples of each of the 21 outbreaks to investigate the nature and extent of diversity. All samples were very closely related to the reference Lordsdale virus genome sequence. LV was first discovered during an hospital outbreak of gastroenteritis in Southampton General Hospital in March 1993. CONCLUSIONS: Noroviruses are a major cause of outbreaks of gastroenteritis in health care settings. LV is the predominant Norovirus in the UK and was detected in outbreaks that occurred during the national peak of gastroenteritis reports in January 2002. PMID- 15135744 TI - Recombinant VP6-based enzyme-linked immunosorbent assay for detection of immunoglobulin G antibodies to Eyach virus (genus Coltivirus). AB - BACKGROUND: Eyach virus (EYAV) is a tick-borne virus belonging to genus Coltivirus, family Reoviridae. It was isolated in Germany and France and has been suspected to be responsible for neurological diseases in humans. To date, there has been no relatively rapid and relatively specific serological assay for EYAV. OBJECTIVES: To develop an ELISA for EYAV, suitable for epidemiological and/or diagnostic purposes. This ELISA should allow to distinguish between infections with EYAV and the related Colorado tick fever virus (CTFV). STUDY DESIGN: VP6, VP7 and VP12 of Eyach virus (the three proteins most divergent between EYAV and CTFV) were expressed in bacteria using the pGEX-4T-2 vector. A partial sequence of VP6 (designated pVP6) was chosen to develop an ELISA for detecting anti-EYAV IgG antibodies in serum. This choice was based on two observations: (i) the homologous VP7 protein of CTFV was successfully used as a target for detecting antibodies to CTFV (the VP7 showed the highest reactivity to an anti-CTFV antibody among all CTFV expressed proteins); (ii) to distinguish infection with EYAV from a CTFV infection: the expressed sequence was chosen within a region which is highly divergent (49% of amino acid identity) from the homologous VP7 sequence of CTFV. RESULTS AND CONCLUSIONS: pVP6 was shown to be the most reactive among the three expressed proteins. The elaborated pVP6 ELISA was evaluated with 340 sera of French blood donors, and found to exhibit a specificity of 100% (no false positives). Furthermore, no cross reaction was detected with antibody to CTFV, thus permitting us to distinguish between infections by either virus. The use of this recombinant protein for serological assays is a good alternative to the use of native EYAV antigen due to the extremely low productivity of the virus in cell culture, and the requirement for suckling mice. This ELISA will be useful to clarify the epidemiological status and the suspected pathogenicity of the virus. PMID- 15135745 TI - Saaremaa hantavirus in Denmark. AB - Most human hantavirus infections in Denmark have been registered on the Fyn island with Puumala hantavirus hosted by Clethrionomys glareolus as the causative agent. To search for other known European hantaviruses in Denmark, we analyzed 101 Apodemus spp. mice trapped on the Lolland island. Genome sequences of Saaremaa hantavirus (SAAV) were recovered from one of the A. agrarius, thus suggesting SAAV presence in Denmark. We also analyzed serum samples from a patient from Lolland with a history of hantavirus-like infection, probably caused by SAAV. Thus, in addition to Puumala virus, SAAV can impose a threat for the human health in Denmark. PMID- 15135746 TI - Comparison of two quantitative CMV PCR tests, Cobas Amplicor CMV Monitor and TaqMan assay, and pp65-antigenemia assay in the determination of viral loads from peripheral blood of organ transplant patients. AB - BACKGROUND: Quantitative PCR assays have become the most common methods in the determination of viral load during cytomegalovirus (CMV) infection of transplant patients. However, usually these tests are still quite time-consuming and labor intensive which diminishes their utility of these tests in routine diagnostic laboratories. OBJECTIVES: The objective of this study was to develop a quantitative CMV PCR test which is time-saving and easy to perform for the detection and monitoring of CMV infection of transplant patients. STUDY DESIGN: The quantitative real time CMV PCR assay using TaqMan chemistry and an automated sample preparation system, MagNA Pure LC, was developed. The designed quantitative CMV test was compared to commercial quantitative PCR test, Cobas Amplicor Monitor, in the determination of CMV DNA loads in plasma samples of liver and kidney transplant patients. The results were also correlated with the CMV pp65-antigenemia test. The clinical material of 270 blood specimens of transplant patients were tested using these two PCR methods and pp65-antigenemia test in parallel. Plasma samples were used for PCR assays and leucocytes for the antigenemia test. RESULTS: The TaqMan assay described was easy to perform, it was rapid (3-4 h) and hands-on time needed for performing the test was short. The detection limit of the assay was 250 copies/ml (cps/ml) plasma and the linear range up to 25,000,000 cps/ml. TaqMan assay was the most sensitive test detecting 92% of the CMV positive findings. Cobas Monitor detected 80% and pp65 test 88% of the positive findings. The correlations between TaqMan and antigenemia assays, and between Cobas Amplicor and antigenemia were statistically significant and high, R = 0.84 (P < 0.0001) and R = 0.80 (P < 0.0001), respectively. Also correlation between two PCR tests was statistically significant (R = 0.64, P < 0.0001). Of the 27 patient studied, 19 demonstrated CMV antigenemia and DNAemia in their blood during the post transplant monitoring. Thirteen of these patients developed a symptomatic CMV infection and were treated with ganciclovir. The peak viral loads of symptomatic patients were statistically higher by all three methods than those of asymptomatic patients. CONCLUSIONS: The developed real time TaqMan assay was rapid and easily performed and could be the best alternative for the diagnosis of CMV infection and monitoring of liver and kidney transplant patients. PMID- 15135747 TI - Human Metapneumovirus as a causative agent of acute bronchiolitis in infants. AB - BACKGROUND: Human Metapneumovirus (hMPV), has been recently isolated from children with acute respiratory tract infections (RTIs), including bronchiolitis, and classified in the Pneumovirinae subfamily within the Paramyxoviridae family. OBJECTIVES: Since most bronchiolitis studies fail to detect any viral pathogen in part of the samples, we sought for the presence of hMPV in a well characterized bronchiolitis cohort. STUDY DESIGN: Nasal washes were obtained from 56 children admitted to the hospital for acute bronchiolitis. RNA extraction and subsequent RT-PCR were used to detect hMPV, and correlated the presence of the virus with clinical characteristics of the disease. RESULTS AND CONCLUSIONS: PCR revealed the presence of hMPV in 16% of bronchiolitis cases, whereas respiratory syncytial virus (RSV; 67.9%) was the most frequently encountered viral pathogen. hMPV was identified either as a unique viral pathogen or co-existed with RSV, with whom they shared a similar seasonal distribution. There were no differences in disease characteristics, either clinical or laboratory, between bronchiolitis cases where hMPV was present and those caused by RSV or other viral pathogens. These findings suggest that hMPV is a common and important causative agent in infants with bronchiolitis, with clinical characteristics similar to that of RSV. PMID- 15135748 TI - Cytomegalovirus UL97 mutations associated with ganciclovir resistance in immunocompromised patients from Argentina. AB - BACKGROUND: Prolonged therapy with ganciclovir (GCV) can result in the development of GCV-resistant strains due to mutations in the viral phosphotransferase (UL97 gene) and/or in the viral DNA polymerase (UL54 gene). OBJECTIVES: The purpose of this study was to detect by molecular methods the most prevalent UL97 mutants which confer ganciclovir-resistance in immunocompromised populations. STUDY DESIGN: Patients from two populations were selected: (a) renal transplant patients with active cytomegalovirus (CMV) infection and more than one cycle of GCV; (b) HIV-infected patients with retinitis due to CMV, who were under GCV induction, maintenance therapy or withdrawal. Patients were followed up by pp65 antigenemia and by viral isolation from blood or/and urine samples. Two fragments (133 and 255pb) of the UL97 gene were amplified by polymerase chain reaction (PCR) from CMV isolates. RESULTS: Nine from 12 isolates obtained were sequenced, three from two renal transplant patients and six from five HIV infected patients. A UL97 mutation, known to confer GCV resistance, was found in two isolates from a renal transplant patient. A methionine to valine mutation at codon 460 (M460V) was detected. These isolates exhibited another mutation at codon 605, whose amino acid changed from aspartic acid (D) to glutamic acid (E). These findings were observed after treatment with IV-GCV/ O-GCV/ IV-GCV for 151 days. The 605 mutation was also detected in leukocytes from the same patient previous to the beginning of the treatment with GCV. CONCLUSIONS: Although a known resistant mutation appeared in a renal transplant patient, it was not associated with CMV disease. We suggest that the D605E mutation could "partially or totally compensate" for the effect of GCV resistance conferred by the 460 mutation. Further studies should be performed to confirm this hypothesis. PMID- 15135749 TI - Modification of CMV DNA detection from dried blood spots for diagnosing congenital CMV infection. AB - BACKGROUND: Detection of viral DNA in dried blood spots using the Guthrie card (DBS test) is a reliable and practical method of diagnosing congenital cytomegalovirus (CMV) infection. The test lends itself to epidemiological studies to establish the prevalence of the infection, but also to neonatal screening for secondary prevention of sequelae. These applications would be facilitated if it were possible to use smaller samples and do the test on pools of individual cases. OBJECTIVE: To ascertain whether doing the test on smaller, pooled samples still accurately identifies neonates with congenital CMV infection. STUDY DESIGN: We tested DBS from: (A) 39 laboratory reference cases; (B) 156 neonates suspected of having congenital CMV infection; (C) 119 children examined for the retrospective diagnosis of congenital CMV; (D) mock specimens prepared with known amounts of viral DNA. RESULTS: The test using only one third of the usual amount of dried blood was 100% sensitive and specific compared to the standard DBS test (A) and to viral isolation (A and B). Pools of three single cases gave the same results as viral isolation (B) and the small-sample test (B and C). All the versions of the test gave a detection limit of 400 copies/ml. CONCLUSIONS: The modified procedure can accurately diagnose congenital CMV infection. It achieves savings in both the patient material and the costs of testing. PMID- 15135750 TI - Long-term follow-up of HIV/HBV coinfected patients from a Greek center. PMID- 15135751 TI - The role of primary care non-physician clinic staff in e-mail communication with patients. AB - BACKGROUND: Few studies have explicitly addressed how e-mail technology may affect non-physician clinic staff, even though these staff typically manage tasks well suited to e-mail communication such as requests for prescription renewals, laboratory and test results, and referral authorizations. GOAL: We conducted a survey of staff members at 10 primary care clinics in Boston to further evaluate non-physician staff attitudes towards e-mail use with patients. We subsequently re-surveyed staff at three of these clinics after the implementation of Patient Gateway, an application designed to facilitate secure electronic communication between patients and the clinics. RESULTS: Before Patient Gateway implementation, 88% of surveyed staff were already using e-mail at least once a day for work related communication. Many of these staff members (24%) were already using e mail with patients. Forty-eight percent of staff members thought that increasing e-mail use with patients could improve the quality of care their practices delivered. However, staff reported having some hesitations about increasing e mail use with patients, mostly relating to security, confidentiality, and workload. After Patient Gateway implementation, users reported high satisfaction with the application and staff in general (users and non-users of Patient Gateway) felt more enthusiastic about increasing e-mail use with patients. CONCLUSIONS: In order to maximize the potential of staff-patient e-mail, it is important that concerns relating to security, confidentiality, and workflow are addressed, and patients must be given guidelines for the appropriate use of e mail. Secure applications designed with these issues in mind are likely to be well received by staff members, and in turn physicians. PMID- 15135752 TI - An evaluation of one-on-one advanced proficiency training in clinicians' use of computer information systems. AB - OBJECTIVE: We examined the effectiveness of a one-on-one training strategy for advanced proficiency in computer information systems (CIS) by clinicians in a large Health Maintenance Organization (HMO). Specifically, this study assessed the level of self-reported improvement in CIS efficiency following one-on-one training, and assessed the perceived value of one-on-one training compared to other teaching methods. DESIGN: We performed a cross-sectional study using a paper-based survey of 129 clinicians practicing in the HMO. MEASUREMENTS: We used a multi-item satisfaction index to measure clinician satisfaction with the one-on one training. We measured whether clinicians thought they were more efficient using the system after training. RESULTS: The one-on-one method was significantly preferred over other teaching methods. Compared to other CIS components, use of the electronic medical record (EMR) improved most following one-on-one training. Sixty-one percent of the clinicians reported major improvements (i.e., >3 on a 5 point Likert scale; five being the highest score) in using the EMR. CONCLUSION: Perceived effectiveness of one-on-one training and overall satisfaction were ranked high by clinicians. The findings support the assumption that clinicians value one-on-one training and value this training method above other methods. PMID- 15135753 TI - Information exchange in the NICU: what sources of patient data do physicians prefer to use? AB - BACKGROUND: There are significant gaps in understanding what sources of patient information physicians utilize in the Neonatal Intensive Care Unit (NICU). OBJECTIVE: We conducted a qualitative study of physicians' self-report of information-seeking behaviors in the NICU of an academic medical center. METHODS: The study used a survey design to assess resident and faculty physicians' perceptions of their utilization of written and verbal sources of patient information. Faculty and resident responses were compared by t-tests to assess how perceptions of information-seeking behavior might differ between these two groups. RESULTS: Of the options listed in our survey, the three most commonly reported information sources were: (1) the bedside flowsheet; (2) conversations with resident physicians; and (3) conversations with nurses. Notes written by physicians-especially resident notes-were the least reported source of patient information. CONCLUSIONS: Physicians' preference to use verbal communication is consistent with prior studies. This study identifies that the bedside flowsheet is also an important source of information, while other written sources especially resident notes-appear not to be utilized as frequently. Understanding why physicians use or fail to use different sources of patient information may shed light on ways to improve information exchange and reduce medical error in complex settings such as the NICU. PMID- 15135754 TI - Barriers to the adoption of computerised decision support systems in general practice consultations: a qualitative study of GPs' perspectives. AB - INTRODUCTION: Computerised decision support systems are increasingly important in primary care for the practice of evidence-based medicine and the development of shared GP-patient decision making. However, despite their emergence, such systems have not been entirely embraced by GPs. There is little qualitative research exploring practical barriers to the adoption of decision support systems in this setting. METHOD: Qualitative interviews with 15 GPs in the West Midlands. RESULTS: Several practical barriers were identified to the use of computerised support systems in primary care consultations. These included limitations of practitioners' IT skills, problems for GPs in understanding the risk output of systems and GP concerns about communicating risk sufficiently well to patients. Concerns over the time implications of using a system in a consultation was also identified as a barrier. CONCLUSION: Designers of decision support systems for use in primary care consultations must account for the practical needs of users when developing computerised support systems. Systems must be acceptable to the format of a consultation, include definitions of what output means, and help facilitate dialogue between the GP and the patient. PMID- 15135755 TI - Information access at the point of care: what can we learn for designing a mobile CPR system? AB - OBJECTIVES: Hospitals have started to migrate their paper-based records to computerized patient records (CPR). The majority of today's CPR systems are stationary, which means that physicians use a clinical workstation to access CPR information. But health care professionals need to request and enter information at different locations, for example, on their daily ward round. This suggests the use of mobile computers, enabling an ubiquitous access to needed data. Different studies show that health care professionals are reluctant to use poorly designed mobile CPR systems, as the work at the point of care is very time-pressured and hectic. To design a system with high acceptance, it is essential to obtain empirical insight into the work practices and context in which the mobile CPR system will be used. METHOD: We investigated the physicians' work with the patient record during their daily round. With the help of a compact notation method, the physicians' interaction with the information system was recorded in real time. Fourteen physicians from three different departments (internal medicine, surgery, and geriatrics) of a middle-sized Swiss hospital participated in our study. RESULTS: Physicians have clear access preferences when they interact with the patient record during their daily round. There exists a clear profile of access frequencies and patterns, respectively. As an example, approximately 50% of all patient record accesses concern information about medications, vital signs and lab test results. DISCUSSION/CONCLUSION: A CPR system which is designed to reflect the access frequencies and patterns should improve the efficiency of data entry and retrieval and thus result in a system with high acceptance among physicians in the demanding environment during hospital rounds. PMID- 15135756 TI - An information system for drug prescription and distribution in a public hospital. AB - With 773 beds and 3696 employees, the Hospital das Clinicas da Faculdade de Medicina de Ribeirao Preto (HCFMRP)-Universidade de Sao Paulo, is one of the largest medical institutions in Latin America. The complete process of prescribing and distributing medication at HCFMRP involves the following stages: prescribing (physicians); ordering (infirmary); separating and dispensing (main drugstore); verifying and administering (infirmary). This was a manual process, normally taking place in the morning. Bottlenecks were inevitable and the risk of errors was elevated. An information system (IS) was devised and implemented with a view to controlling such problems. This article addresses the construction of this system and reports on a survey in which different groups of users have evaluated the project. PMID- 15135757 TI - Development and evaluation of an integrated pharmaceutical education system. AB - There is increasing evidence that patient safety can be improved by the introduction of an integrated computer-based medical care system in hospital settings. In this paper, we describe an integrated pharmaceutical information system (IPIS) in which a patient's profile including his/her medication records and prescriptions are collected from physician order entry systems and pharmaceutical systems along with the history of patient care in the hospital. Based on an individual patient's profile the IPIS can provide pharmaceutical education information specifically to meet the patient's needs. The IPIS has been developed and installed at Taipei Medical University Wanfang Hospital (TMUWFH) since July 2002. Evaluation of the system showed that it can help patients to effectively acquire drug information. This enables them to have a much better understanding of the pharmacological properties of the medicines they are taking, including adverse drug reactions and side-effects. In our opinion the system has the potential to improve both patient safety and treatment outcomes. PMID- 15135758 TI - Allied health professionals' use of online evidence: a survey of 790 staff working in the Australian public hospital system. AB - OBJECTIVE: To measure the impact of a state health department policy to provide allied health professional staff with access to a point-of-care, 24h, online evidence system in terms of awareness, use and clinical impact of the system on clinical practice; to identify perceived barriers to use, and differences in measures between seven professional groups (physiotherapists, occupational therapists, speech pathologists, dieticians, clinical psychologists, pharmacists and social workers). METHOD: A convenience sample of 790 allied health professionals from 65 randomly selected hospitals in New South Wales, Australia. A self-administered questionnaire was distributed. Rates of use, frequency and types of resources accessed were calculated. Comparisons between professional groups were undertaken using Chi-square analyses and t-tests. RESULTS: The results showed that 82% of allied health professionals had heard of the online evidence system, and of those 76% had used it. Pharmacists had the highest rates of use and social workers the lowest. Of users, 90% agreed that use of the system had the potential to improve patient care and 45% reported direct experience of this. Computer skills and easy access were significantly associated with use and frequency of use. Among non-users, lack of specific training in the use of the online evidence system and lack of time were the most frequently reported reasons for not using the system. However, among users there was no relationship between this training and the frequency or effectiveness of use, i.e. the ability to find the information required. CONCLUSIONS: Allied health professionals will use an online evidence system when it is provided, however there are marked differences in use by professional groups. General training aimed at improving computer skills appears more important in encouraging use of an online evidence system, than specific system-based training. Perceptions of organisational and professional support for allied health professionals to use online evidence as a legitimate part of their work play an important role in influencing system use. PMID- 15135759 TI - Viewpoint--midwives' enthusiasm for complementary therapies: a cause for concern? PMID- 15135760 TI - Complementary medicine and older people: past research and future directions. AB - Due to the natural aging processes, older people are particularly susceptible to a range of chronic health conditions. However, despite that research has indicated that chronic health conditions and disability act as reliable predictors of complementary/alternative medicine (CAM) use, despite research evidence that older people are significant consumers of CAM, and regardless of the potential for CAM to enhance successful aging, reduce frailty, and increase independence and quality of life in older persons, older people's use of CAM therapies remains under-researched. This paper reviews what existing research literature is there on CAM use in older age; considers rates of and potential for use, features of and barriers to use, and economic, effectiveness and safety issues. From these beginnings, outlined is a wide-ranging research agenda on CAM and older people. PMID- 15135761 TI - Finding the right complementary therapies course. AB - Over the past 8 years the number of full time, Complementary Therapies degree courses provided within UK Universities has increased from less than five to currently almost one hundred. In addition, the developments within the Further Education and private sector have added to the increase in provision. With so many different courses on offer, finding the most suitable, quality course is becoming a challenge for applicants. Course titles provide little insight into the philosophy, nature and quality of provision while details within an institute's prospectus or web site frequently do not clearly identify distinguishing features. Key characteristics of quality provision to look for in a course in complementary therapies are discussed. Aspects considered include: the type of institute, modes of study, which therapy and its current regulation, the skills development and clinical experience opportunities, the balance of subjects within a course, the experience of the teaching team and their research activities. Discussion is included of the need for adequate quality assurance procedures to ensure courses evolve to continue to meet the needs of those entering the rapidly developing profession. The relationship of the provider and the appropriate professional bodies is also discussed. A summary of aspects to consider when looking for a comprehensive, quality course has been provided in Table 2. PMID- 15135762 TI - The clinical feasibility of natural medicine, venotonic therapy and horsechestnut seed extract in the treatment of venous leg ulceration: a descriptive survey. AB - BACKGROUND: The primary treatment of choice for venous leg ulceration (VLU) is compression therapy; however, serious clinical issues demand the development of new treatments. An extract believed to promote VLU healing is Horsechestnut Seed Extract (HCSE). METHODS: The clinical feasibility of HCSE in VLU was explored in a two-stage design. The second stage presented here, was a descriptive survey exploring current opinion and utilisation of natural therapies, venotonics and HCSE in VLU. A questionnaire mailed to 122 district nurses, 73 medical practitioners (MPs) and 53 clients with VLU resulted in a response rate of 32, 31.5 and 81 per cent, respectively. RESULTS: Natural therapy and HCSE use for VLU was minimal in all groups. Half of all groups supported venotonics, with a similar proportion of nurses and clients utilising venotonics in practice. MPs were however less likely to utilise venotonics for VLU. CONCLUSION: Whilst clinicians indicate that clinical evidence may influence the utilisation of HCSE in clinical practice, the evidence currently does not exist. Positive findings from well-designed trials may ameliorate the integration of natural medicine into mainstream practice. PMID- 15135763 TI - Complementary medicine at Exeter: the first 10 years. PMID- 15135764 TI - An aura of confusion Part 2: the aided eye--"imaging the aura?". AB - The second of three papers considers claims made for the perception or detection of vital energy. Many systems of complementary and alternative medicine assume the existence of a vital force that mediates therapeutic efficacy, for example chi or qi in Traditional Chinese Medicine. Vital energy directly perceived or imaged that surrounds living organisms is frequently termed the aura. A range of devices or techniques are claimed to image the aura. These include Kirlian photography, gas discharge visualisation (GDV) and polychromatic interference photography (PIP). This paper explores such claims and argues that the images produced can be explained using concepts from the physical sciences. It is suggested that techniques such as KP, GDV or PIP currently offer insufficient reliable research evidence concerning their use as diagnostic or imaging alternatives. Consequently their clinical use is debatable. Kirlian photography and its derivatives may however be useful as a research tool by providing visual records of complex bodily responses to experimental situations. For example, responses to physiological or psychological stressors. PMID- 15135765 TI - The award winning "Completing the circle" complementary therapy service initiative at Christie NHS Hospital Manchester. AB - The complementary therapy team at the Christie Hospital has won a highly prestigious national Prince of Wales award. The award is for "Good Practice in Integrated Healthcare" and applauds the hospital for including complementary therapies as part of patient care at the Christie. His Royal Highness, The Prince of Wales presented The Prince of Wales Foundation for Integrated Healthcare Award to Christie staff at Lambeth Hospital in London on the 11th December 2003 (see picture). The Gateway Clinic at Lambeth Hospital jointly won the award (reference here to a future publication). The Christie Hospital started providing complementary therapies 7 years ago, and the service has greatly expanded in the last 3 years. Patients can choose a complementary therapy treatment in addition to their medical treatment. A 15-strong complementary therapy team provides back and head massages, reflexology and other treatments. The paper reports on the various aspects of the project, identifying who leads each part and how it is being developed and evaluated. PMID- 15135766 TI - Proceedings of the 35th International Symposium on Growth Hormone and Growth Factors in Endocrinology and Metabolism. April 4-5, 2003. Prague, Czech Republic. PMID- 15135768 TI - Growth and growth hormone in children born small for gestational age. AB - At present, there are no uniform definitions for terms such as "small for gestational age" (SGA) and "catch-up growth" that can be used when evaluating children who are born small. Data suggest that growth hormone (GH) therapy can play a role in helping children born SGA achieve adequate height. For therapy to be successful, however, precise terminology must be defined and standardized, surveillance and identification of potential candidates must be performed, and treatment plans must be tailored to the unique needs of each patient. This manuscript focuses on each of these issues, and also identifies additional concerns that must be addressed as a consequence. PMID- 15135769 TI - Intellectual performance in young adult males born small for gestational age. AB - Children born small for gestational age (SGA) have increased risk of neurological dysfunction. The aim of this study was to analyze intellectual performance in early adult age in children born SGA, and especially the effect of catch up growth in height. METHODS: Data on gestational age and birth size were taken from the Swedish Birth Register and data on adult height and intellectual performance from the Swedish Conscript Register. RESULTS: Low birth weight, birth length and head circumference increased the risk of subnormal intellectual and psychological performance in males born SGA. Very low gestational age also increased the risk. However, the most important predictor was the absence of catch up growth to a height above -2 SDS in individuals born SGA. PMID- 15135771 TI - Reciprocal interactions between the GH axis and sleep. AB - For more than 30 years, growth hormone (GH) has been observed to be preferentially secreted during deep, slow-wave sleep (SWS). However, the mechanisms that underlie this robust relationship that links anabolic processes in the body with behavioral rest and decreased cerebral metabolism remain to be elucidated. Current evidence indicates that GH secretion during the beginning of sleep appears to be primarily regulated by GH-releasing hormone (GHRH) stimulation occurring during a period of relative somatostatin withdrawal, which also is associated with elevated levels of circulating ghrelin. Apparently, two populations of GHRH neurons need to be simultaneously active to stimulate, in a coordinated fashion, SWS and pituitary GH release. Pharmacological interventions that are capable of increasing the duration and/or the intensity of SWS such as oral administration of gamma-hydroxybutyrate (GHB), also increase the rate of GH release. Because the normal negative feedback exerted by GH on central GHRH is inoperative in patients with GH deficiency, it is possible that the decreased energy levels and fatigue often reported by GH-deficient adults partly reflect an alteration in sleep-wake regulation. Preliminary data from a sleep study of adults with GH deficiency using wrist actigraphy for 6 nights at home and polysomnography in the laboratory indeed show decreased total sleep time and increased sleep fragmentation in GH-deficient patients as compared with normal controls. PMID- 15135772 TI - Local synthesis and dual actions of progesterone in the nervous system: neuroprotection and myelination. AB - Progesterone (PROG) is synthesized in the brain, spinal cord and peripheral nerves. Its direct precursor pregnenolone is either derived from the circulation or from local de novo synthesis as cytochrome P450scc, which converts cholesterol to pregnenolone, is expressed in the nervous system. Pregnenolone is converted to PROG by 3beta-hydroxysteroid dehydrogenase (3beta-HSD). In situ hybridization studies have shown that this enzyme is expressed throughout the rat brain, spinal cord and dorsal root ganglia (DRG) mainly by neurons. Macroglial cells, including astrocytes, oligodendroglial cells and Schwann cells, also have the capacity to synthesize PROG, but expression and activity of 3beta-HSD in these cells are regulated by cellular interactions. Thus, Schwann cells convert pregnenolone to PROG in response to a neuronal signal. There is now strong evidence that P450scc and 3beta-HSD are expressed in the human nervous system, where PROG synthesis also takes place. Although there are only a few studies addressing the biological significance of PROG synthesis in the brain, the autocrine/paracrine actions of locally synthesized PROG are likely to play an important role in the viability of neurons and in the formation of myelin sheaths. The neuroprotective effects of PROG have recently been documented in a murine model of spinal cord motoneuron degeneration, the Wobbler mouse. The treatment of symptomatic Wobbler mice with PROG for 15 days attenuated the neuropathological changes in spinal motoneurons and had beneficial effects on muscle strength and the survival rate of the animals. PROG may exert its neuroprotective effects by regulating expression of specific genes in neurons and glial cells, which may become hormone-sensitive after injury. The promyelinating effects of PROG were first documented in the mouse sciatic nerve and in co-cultures of sensory neurons and Schwann cells. PROG also promotes myelination in the brain, as shown in vitro in explant cultures of cerebellar slices and in vivo in the cerebellar peduncle of aged rats after toxin induced demyelination. Local synthesis of PROG in the brain and the neuroprotective and promyelinating effects of this neurosteroid offer interesting therapeutic possibilities for the prevention and treatment of neurodegenerative diseases, for accelerating regenerative processes and for preserving cognitive functions during aging. PMID- 15135774 TI - Growth hormone insensitivity resulting from post-GH receptor defects. AB - Biochemical analysis indicates that the STAT-5b mutation affects signaling by both growth hormone (GH) and gamma-interferon. A patient with such a mutation thus manifests two new clinical disorders: (1) growth hormone insensitivity (GHI), which results from a post-receptor defect in GH signaling and (2) a new form of primary immunodeficiency. Given that the GH receptor is a member of the hematopoietin-receptor family, it seems reasonable to predict that additional cases of defects in GH signaling will be identified. The predicted phenotype would be GHI combined with defects in the immune system. PMID- 15135775 TI - Role of serum insulin-like growth factor I in mammalian brain aging. AB - Modern societies face new public health challenges associated with an increasingly aging population. Among these, pathological conditions linked to brain aging are paramount. Old age is a risk factor for important neurological impairments such as Alzheimer's disease or stroke. Even healthy elderly people usually present with milder forms of cognitive decline. This is possibly related to less-pronounced brain deficits seen in normal aging, including the shrinkage of neurons and the dense network of neurons and glia in the central nervous system known as the neuropil, a lower neurogenetic rate, impaired angiogenesis or brain accumulation of deleterious compounds. At least in mammals, age is also associated with a decline in insulin-like growth factor-I (IGF-I) levels, a well known neuroprotective agent. Recently, a relationship between serum IGF-I and "house-keeping" mechanisms in the brain has been evidenced in laboratory rodents. Serum IGF-I increases adult neurogenesis, sustains neuronal health through a variety of fundamental homeostatic mechanisms, participates in brain angiogenesis, contributes to brain beta-amyloid clearance and affects learning and memory. Overall, diminished trophic input resulting from decreasing serum IGF I levels during aging likely contributes to brain senescence in mammals. PMID- 15135777 TI - Molecular mechanisms of end-organ resistance. AB - Hormone resistance is caused by mutations in hormone receptors or by functional desensitization of hormone signaling pathways. Mutations that cause hormone resistance have been described for most classes of hormones, including peptides like growth-hormone releasing hormone (GHRH); large proteins such as insulin and the glycoprotein hormones; steroid hormones; and even ions such as calcium. In the case of proteins, many of these receptors include seven transmembrane, G protein-coupled receptors such as those for thyroid-stimulating hormone (TSH), luteinizing hormone (LH), follicle-stimulating hormone (FSH), adrenocorticotrophic hormone (ACTH), GHRH and vasopressin. However, other classes of membrane receptors can also be affected. For example, the insulin receptor is a tyrosine kinase receptor, the growth hormone (GH) and leptin receptors belong to the cytokine family of receptors, and the Mullerian inhibiting substance (MIS, also known as anti-Mullerian hormone [AMH]) receptor is related to the transforming growth factor beta (TGFbeta) serine kinase group of receptors. G protein mutations cause Albright's hereditary osteodystrophy, and, as might be expected, there is resistance to many different hormones that act through G protein-coupled receptors. A variety of resistance syndromes involve members of the nuclear receptor superfamily, including resistance to androgens, vitamin D, thyroid hormone, glucocorticoids and estrogen. Mutations in orphan nuclear receptors such as steroidogenic factor-1 (SF-1) or DAX-1 result in defective glandular development, thereby indirectly causing hormone resistance. Another form of resistance, sometimes referred to as "post-receptor" resistance, is typified by acquired forms of insulin or leptin resistance, and the pathophysiology remains elusive. Studies of naturally occurring mutations have provided important insights into the structure and function of receptors. By definition, the mutations that cause disease identify important functional domains in the proteins. Although therapy is straightforward in some disorders, it is challenging or incompletely studied in others. PMID- 15135778 TI - Aspects of growth hormone deficiency and replacement in elderly hypopituitary adults. AB - Normal ageing is associated with a decline in spontaneous growth hormone (GH) secretion, and although elderly hypopituitary adults demonstrate an increase in total and central fat compared with age-matched controls and are distinguishable from control subjects in terms of GH responsiveness on dynamic testing, there are few data available on the response to GH replacement in older subjects. We have studied the baseline characteristics of 295 patients (173 males and 122 females) aged >65 years of age who began GH replacement therapy at the time of entry into the KIMS program (Pfizer International Metabolic Database) and the effects of GH replacement in 125 patients who completed at least 12 months of GH replacement therapy. Data were compared with those of 2469 (1249 males and 1220 females) patients aged <65 years with adult-onset GH deficiency (GHD). The patients were selected using strict criteria in accordance with the recommendations from the Growth Hormone Research Society. There was a higher proportion of pituitary adenoma relative to craniopharyngioma in the older age group (P<0.001), but there was no difference between groups in the degree of hypopituitarism (number of additional hormone deficiencies). Blood pressure, cholesterol and low-density lipoprotein (LDL) cholesterol levels were positively correlated with age, and older patients had a predictably higher prevalence of diabetes mellitus, coronary heart disease, stroke and history of hypertension. Quality of life (Assessment of Growth Hormone Deficiency in Adults (AGHDA) score) was impaired in both groups before the start of GH therapy. GH replacement doses were lower in older patients with GHD as compared with patients <65 years old. After 12 months of GH replacement, significant improvements were evident in waist circumference, waist/hip ratio, lean body mass, diastolic blood pressure, total and LDL cholesterol levels and AGHDA scores in patients aged <65 years. Similar significant reductions were evidenced in patients >65 years old compared with those observed in younger patients. The total number of adverse events was similar in younger and older patients with GHD. However, younger patients had more fluid retention-related adverse events such as headache, oedema and arthralgia; whereas, older patients with GHD had more adverse events related to glucose metabolism, cardiovascular events and neoplasms. These data indicate a positive benefit from GH replacement in older patients with hypopituitarism - particularly in relation to quality of life - using a lower dose of GH for replacement and with appropriate age-related safety controls. PMID- 15135780 TI - Congenital adrenal hyperplasia: transitional care. AB - Congenital adrenal hyperplasia (CAH) is a life-long disorder which poses management problems that are age- and sex-specific. The condition merits an organised, multi-disciplinary transitional care format similar to the kind that is now well established for Turner's syndrome in many centres. In the eyes of the paediatrician, achieving optimal growth is the primary target of CAH management during infancy and childhood. Fixation on this objective can be to the detriment of the patient because it may result in failure to appreciate the significance of metabolic disturbances that occur in later childhood, particularly in females, and which may be the progenitor of chronic problems with obesity, insulin resistance and infertility in adult life. Similarly, the care of the adult patient with CAH comprises more than just prescribing steroid replacement for primary adrenal insufficiency. The transition period between childhood and adulthood is an opportune time for review of the various management options and to assess the efficacy of steroid replacement, to consider alternative novel treatment modalities and to apply a checklist to the multi-faceted aspects of the medical, surgical and psychological needs of the patient. PMID- 15135781 TI - Congenital adrenal hyperplasia due to 21-hydroxylase deficiency--the adult woman. AB - Congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency is a disease with a varying phenotype depending on the mutation(s) present and the severity of the disease. All children with CAH need to be continuously cared for from birth or early infancy by specialists in paediatric endocrinology and surgery. Complications due to over- or under-treatment with corticosteroids are often seen during adolescence, and these problems often continue into adulthood. For the young woman with CAH, questions about menstruation, sexuality, fertility and the possible necessity of complementary surgery are always important issues that need to be discussed. To meet the needs of the young woman with CAH, it is important that the transition from paediatric to adult care be a process of parallel consultations over several years, always involving an experienced gynaecologic endocrinologist. PMID- 15135782 TI - Transition in Turner's syndrome. AB - Management of the chromosomal condition Turner's syndrome requires consistent medical care, especially during the time when affected girls transition from childhood into adulthood. The medical problems that first develop during childhood of a patient with Turner's syndrome such as congenital heart disease, hearing loss, skeletal problems and dental and ophthalmological abnormalities, should be followed into adulthood. Providing the necessary continuum of care will require that medical centers develop teams with the appropriate expertise in treatment of Turner's syndrome. Now more than ever patients with Turner's syndrome have the capability of achieving their full potential, but it requires a multidisciplinary approach toward care throughout their lifetime. PMID- 15135783 TI - Considerations for transition from paediatric to adult endocrinology: women with Turner's syndrome. AB - Turner's syndrome (TS) is a genetic anomaly that results from complete or partial absence of one X chromosome and is the most commonly occurring chromosomal abnormality in females. While most females with TS present at birth or in early childhood, nearly one quarter present at adult services with primary or secondary amenorrhoea. It is only with the advent of clinics dedicated to adults with TS that we have realised the degree of occult pathology present in this group. Adults with TS are thought to have a reduced life expectancy, mainly due to excess cardiovascular risk, but they may also have multiple comorbidities including hypothyroidism, deafness, osteoporosis and the attendant problems of oestrogen deficiency and infertility. Many of these features may be affected by the timing of treatments in paediatrics - particularly the timing of growth hormone (GH) and oestrogen use. It is the role of adult TS services to audit the outcome of TS and to inform their paediatric colleagues of the long-term effects of childhood treatments. Thus, a multidisciplinary approach to focused adult care is needed, with consideration of how to optimise surveillance strategies in these women. PMID- 15135784 TI - Transition from paediatric to adult endocrinology: hypopituitarism. AB - For patients at the transition stage between childhood and adulthood, cooperation between paediatric and adult endocrinologists is essential for optimum care. Evaluations of patients in transition consist of retrospective assessments of what happened in childhood combined with prospective planning for treatment and follow up into adult life. Successful transfer of patients in transition from paediatric to adult care should follow a general sequence tailored to local circumstances. First, all patients with GH deficiency and their families should be informed by their paediatric endocrinologist about the long term consequences of GH deficiency in adulthood and the potential need for life-time GH replacement therapy. Second, retesting will be needed for evaluating hypothalamic pituitary function, re-evaluation of the need for replacement therapy for other pituitary hormone deficits, measurement of fasting lipid concentration and assessment of skeletal integrity. Third, the patient and physicians should consider the option of attending an out-patient clinic having both a paediatric and an adult endocrinologist. Regardless, the wishes of the patient should be respected as much as possible. It is essential that those patients who had childhood-onset GH deficiency and who were treated with GH be followed throughout adult life. PMID- 15135786 TI - Familial acromegaly. AB - Most pituitary tumors are sporadic, though a few occur with a familial aggregation. Three distinct syndromes have been recognized to date: multiple endocrine neoplasia, type I (MEN-1), Carney complex (CNC), and isolated familial somatotropinomas (IFS). Pituitary tumor types in MEN-1 are similar to those occurring sporadically. The largest percentage are prolactin-secreting or non functioning and only about 10% are growth hormone (GH)-secreting (somatotropinomas). In contrast, tumors types in CNC and IFS are invariably somatotropinomas, though there are differences in both clinical and histological features. Each of the familial syndromes is associated with a tumor-suppressor gene that was initially recognized by an observed loss of heterozygosity on chromosome 11q13 in MEN-1 and IFS and on chromosome 17q in CNC. The MEN-1 gene, which codes for the nuclear protein, menin, has been identified and a large number of inactivating mutations have been recognized. The gene associated with CNC codes for the protein kinase A regulatory subunit 1, inactivation of which leads to enhanced activity of the GH-releasing hormone-induced signal transduction pathway. This pathway exerts proliferative effects in somatotropes. The gene associated with IFS is distinct from the MEN-1 gene, though it is located in close proximity, and is contained in a candidate region of approximately 10 Mb. Identification of the IFS gene should provide new insight into the pathogenesis of somatotropinomas, not only in IFS but also in sporadic tumors, where there is an up to 40% allelic loss on chromosome 11q13. PMID- 15135787 TI - Biochemical markers of acromegaly: GH vs. IGF-I. AB - The development of sensitive and specific growth hormone (GH) and insulin-like growth factor I (IGF-I) assays opened a new page in the diagnosis and surveillance of acromegaly. Currently, it is possible to make an accurate pre operative diagnosis even in patients with virtually no typical clinical signs of the disease and to monitor the efficacy of therapeutic intervention with a high degree of precision. This review briefly discusses the performance parameters of GH and IGF-I as diagnostic and surveillance tools in patients with acromegaly. In brief, whereas GH-based parameters may offer the advantage of disclosing dysregulation of GH secretion, a single plasma IGF-I measurement provides the most comprehensive assessment of both the overall GH output as well as the pattern of GH presentation to the peripheral tissues. Judicious use of both biochemical markers allows accurate and early diagnosis, precise assessment of the efficacy of therapeutic interventions, permits adjustment in the dose of medication and allows for unequivocal demonstration of a final cure. PMID- 15135788 TI - The place of pegvisomant in the acromegaly treatment algorithm. AB - The disfiguring disease acromegaly results from hypersecretion of growth hormone (GH). The main goals of treatment for acromegaly include normalisation of biochemical markers of disease activity to restore normal life expectancy, amelioration of signs and symptoms of the disease, removal of the pituitary tumour without damaging the optic chiasm and other peripituitary structures, and preservation of pituitary function. Conventional options for treatment of acromegaly include surgery, radiotherapy (RT), and medical therapy with either dopamine agonists or somatostatin (SMS) analogues. The advent of the genetically engineered growth hormone analogue pegvisomant is unlikely to alter significantly the place of surgery and RT in the treatment algorithm for acromegaly biochemical control as determined based on serum IGF-I concentrations is achievable with pegvisomant in virtually all patients, and it will clearly become the drug of choice in patients partially or completely unresponsive to SMS analogues. Preliminary studies suggest improved insulin sensitivity for a given IGF-I with pegvisomant compared with SMS analogues; if these results are confirmed by results of future studies, such a metabolic advantage may encourage the use of pegvisomant. PMID- 15135790 TI - Rehabilitation and hypopituitarism after traumatic brain injury. AB - Traumatic brain injury (TBI) is a leading cause of death and disability in the United States (US). The severity of a TBI is difficult to assess, and therefore, accurate determination of the prognosis is difficult. The symptoms of a TBI involve most major medical systems, and share many similarities with the symptoms of hypopituitarism. Although more than two-thirds of patients with severe head injuries who die have structural abnormalities in the hypothalamus and the pituitary, pituitary function is not routinely assessed after head trauma. Thus, studies have shown that anterior pituitary hormone deficiencies are common following brain injury [Endocrinologist 11 (2001) 275; J. Clin. Endocrinol. Metab. 86 (2001) 2752]. Survivors of brain injury should be screened for such deficiencies so that replacement therapy can be initiated to optimize rehabilitation and outcome. PMID- 15135791 TI - Hypopituitarism and growth hormone deficiency (GHD) after traumatic brain injury (TBI). AB - In adults, hypopituitarism and growth hormone deficiency (GHD) should be suspected and diagnosed within an appropriate clinical context. It has been demonstrated that all patients with primary hypothalamic-pituitary diseases before and after any medical intervention (defined as neurosurgery, radiotherapy and medical therapy) are at obvious risk - more than just at high risk - for hypopituitarism (greater than 80% had severe GHD). The same obvious risk applies to patients diagnosed as having congenital or acquired GHD in childhood (between 30% and 50% of patients with severe GHD after retesting). Taking into account the fragility of the infundibular-hypothalamic structure, patients with other common pathological conditions of the central nervous system (CNS), such as traumatic brain injury (TBI), subarachnoid haemorrhage (SAH) or primary brain tumours (BT) with related medical intervention could be at risk for developing hypopituitarism, including GHD. GHD is the first and most common sign of pituitary impairment. Despite the risk of pituitary dysfunction and the results of some studies that these risks are greater than previously believed, neuroendocrine evaluations are still not routinely included as part of the clinical management plan for patients with TBI and SAH. Preliminary results of a multicenter study performed under the auspices of the Italian Society of Endocrinology underscore the high risk of hypopituitarism and GHD under these clinical conditions. Thus, careful screening of pituitary function should always be performed in patients following TBI and SAH. PMID- 15135792 TI - Endocrine consequences of brain irradiation. AB - Cranial radiation is routinely used to manage pituitary tumours, craniopharyngiomas, primary brain tumours, tumours of the head and neck and, in the past, for the prophylaxis of intracranial disease in patients with acute lymphoblastic leukaemia. If the hypothalamic-pituitary axis falls within the radiation fields, the patient is at risk of developing hypopituitarism. The effect of radiation is determined by the dose and the time that has elapsed since treatment. Classically, growth hormone (GH) is the most sensitive of the anterior pituitary hormones to irradiation, followed by gonadotrophins, adrenocorticotrophic hormone (ACTH) and thyroid-stimulating hormone (TSH). Low dose irradiation in prepubertal children can initially cause early or precocious puberty and subsequently gonadotrophin deficiency. Higher doses may cause gonadotrophin deficiency and pubertal delay. The ACTH and TSH axes are relatively resistant to the effects of irradiation, but minor abnormalities may occur. Patients who receive cranial irradiation that affects the hypothalamic-pituitary axis remain at risk of developing multiple hormone deficiencies for many years and require long-term follow-up by an endocrinologist. PMID- 15135793 TI - Screening for late effects in survivors of childhood cancer: growth hormone deficiency from a pediatric oncologist's point of view. AB - At the Emma Kinderziekenhuis/Academic Medical Center in Amsterdam, survivors of childhood cancer are screened annually or biennially for the occurrence of late treatment effects. The screening procedures are based on previously used treatment modalities. The data gathered at the outpatient clinic are registered in the database PLEKsys. Evaluation of the data concerning over 1000 cancer survivors screened since the start of the clinic once more illustrated the relation between cranial irradiation and the development of central endocrine abnormalities. Surprisingly, at least a proportion of the growth hormone (GH) deficient cancer survivors were registered as not being on a replacement therapy regimen. The reasons for survivors not to be on replacement therapy are currently being evaluated. The late-effects outpatient clinic and the PLEKsys database provide a platform for additional research in fields including endocrinology, which should be aimed at improving the care for and the health status of the survivors of childhood cancer. PMID- 15135795 TI - The endocrine consequences for very low birth weight premature infants. AB - The aims of this study were: (1) to determine whether premature and small-for gestational-age (SGA) children have alterations to the insulin-like growth factor (IGF)-IGF binding protein axis and (2) to evaluate growth in premature children. Three groups of children were evaluated: (i) premature children of 36 weeks gestation, which included AGA and SGA subgroups; and (iii) children born at term and AGA with normal childhood heights and weights. Fasting plasma IGF-I, insulin-like growth factor binding protein-3 (IGFBP-3), and IGF-II (all expressed as microgm/L) were drawn on available subjects. To examine the influence of SGA on the IGF-IGFBP axis, term SGA subjects were compared with term AGA subjects. To examine the influence of prematurity on the IGF-IGFBP axis, preterm SGA subjects were compared with term SGA subjects and preterm AGA subjects were compared with the normal-stature AGA controls. This ensured that groups of very similar stature and nutritional statuses were compared. Auxological data were available for 24 premature children, and biochemical data were available for 77 children, including the premature children. Across the height standard deviation score (SDS) range, premature children did not reach mid-parental height (MPH) SDS and were approximately 0.6 standard deviations (SDs) below the MPH SD (P < 0.0001). Plasma IGF-I and IGFBP-3 levels were higher in term SGA subjects compared with term AGA subjects (P < 0.001, respectively). Conversely, IGF-I and IGFBP-3 values were lower in the premature SGA subgroup compared with the premature AGA subgroup (P < 0.001 for both), and both were also lower in the premature AGA subgroup compared with the normal-statured AGA subgroup (P < 0.001 for both). IGF-II values were higher in the preterm group than in the term group (P < 0.001). In conclusion, very low birth weight (VLBW) children, regardless of whether they were AGA or SGA, have low plasma IGF-I and IGFBP-3 levels in mid-childhood, suggesting partial growth hormone (GH) resistance. Conversely, term SGA children have elevated plasma IGF-I and IGFBP-3 levels. When combined, premature birth plays a more dominant role than SGA on the IGF-IGF binding protein axis. PMID- 15135796 TI - The metabolic consequences of prematurity. AB - An association between low birth weight, commonly a reflection of an adverse in utero environment, and the subsequent development of diseases such as type 2 diabetes and hypertension in later life is now generally accepted - as is an association between an adverse perinatal environment and a permanent reduction in insulin sensitivity. This and other metabolic abnormalities have been demonstrated from childhood through to adulthood in subjects who were born full term but small for gestational age (SGA). Less is known about children born prematurely into an adverse neonatal environment. We present data demonstrating that premature infants also have metabolic abnormalities similar to those observed in full-term, SGA children, and that these occur irrespective of whether the premature infants are SGA or appropriate for gestational age (AGA). PMID- 15135797 TI - Pathogenesis of retinopathy of prematurity. AB - Retinopathy of prematurity (ROP) is a major cause of blindness in children in developed countries. ROP is a two-phase disease, beginning with delayed retinal vascular growth after premature birth (Phase I). Phase II follows when Phase I induced hypoxia releases factors to stimulate new blood vessel growth. Both oxygen-regulated and non-oxygen-regulated factors contribute to normal vascular development and retinal neovascularization. Vascular endothelial growth factor (VEGF) is an important oxygen-regulated factor. A critical non-oxygen-regulated growth factor is insulin-like growth factor-I (IGF-I). In knockout mice, lack of IGF-I prevents normal retinal vascular growth, despite the presence of VEGF, important to vessel development. In vitro, low IGF-I levels prevent VEGF-induced activation of Akt, a kinase critical for vascular endothelial cell survival. We found that premature infants who develop ROP have low levels of serum IGF-I compared to age-matched infants without disease. IGF-I is critical to normal vascular development. Low IGF-I predicts ROP in premature infants, and restoration of IGF-I to normal levels might prevent ROP. PMID- 15135799 TI - Integration of genetics into medical practice. AB - It has been a century since the first human genetic disorders were recognized, but only recently have there been any prospects that the genetic approach would become integral to medical practice. Throughout most of the 20th century, medical genetics has focused on rare monogenic and chromosomal disorders. There were major successes, including chromosomal analysis, prenatal diagnosis and newborn screening for inborn errors of metabolism, but the impact was confined to a relatively narrow corner of medicine. The situation has changed, however, with advances in genetics and especially with the sequencing of the human genome. The tools are now at hand to begin to understand the genetic basis of common as well as rare disorders. It is expected that this will lead to major advances in both diagnosis and treatment, so that physicians in all areas of medicine will be using the tools of genetics in their daily practice. PMID- 15135800 TI - Genetic testing in endocrinology: lessons learned from experience with multiple endocrine neoplasia type 2 (MEN2). AB - Multiple endocrine neoplasia type 2 (MEN2) is a syndrome characterized by medullary thyroid carcinoma (MTC), unilateral or bilateral pheochromocytoma and hyperparathyroidism. Familial MTC (FMTC) is a subvariant of MEN2 in which affected individuals develop MTC without other manifestations of MEN2. The identification of RET proto-oncogene mutations in MEN2 and FMTC have provided a precise method for identifying gene carriers. This review provides a concise discussion of the use of genetic testing in the management of hereditary MTC, discussing the appropriate use of this new technology with an emphasis on early intervention to prevent death or serious morbidity from this disease. PMID- 15135801 TI - Fragile X mental retardation syndrome: from pathogenesis to diagnostic issues. AB - The Fragile X (FRAXA) syndrome is the most common cause of familial (monogenic) mental retardation and is widespread in human populations. This syndrome is characterised by an unusual mode of transmission for an X-linked disease. In affected families, one frequently finds clinically normal transmitting males, whose daughters - also clinically normal - have a high risk of having affected children. The risk of developing the disease (penetrance) thus appears to increase in successive generations of the same family through maternal transmission. As shown by molecular cloning of the fragile X locus, Fragile X mutations are unstable expansions of a CGG trinucleotide repeat, located in the first exon (non-protein-coding) of the FMR1 gene (for Fragile X Mental Retardation). Two main types of mutation are observed in affected families. A full mutation is found in patients with mental retardation and corresponds to large expansions of the repeat. Premutations are moderate expansions and are found in normal transmitting males and in the majority of clinically normal carrier females. About 15% of patients show a mosaic pattern consisting of both full mutations and premutations. Although analysis of the CGG expansion has led to the establishment of reliable tests for diagnosis and genetic counseling of Fragile X syndrome, care must be exercised to use these tools to answer the concerns of the families and avoid doing harm. In our opinion, testing in children should be restricted to those who show a developmental delay, cognitive deficits and/or abnormal behavior evocative of the syndrome. A carrier diagnosis in a girl who is clinically normal should probably only be performed at an age where she can understand the consequences for family planning and the options of prenatal diagnosis. When testing children with borderline cognitive deficits, a positive diagnosis should be used to improve educational strategies for the children - and not to stigmatise them. PMID- 15135802 TI - In vitro effect of indomethacin and interferon-alpha on Th1 and Th2 cytokine synthesis in patients with chronic hepatitis C. AB - Current evidences suggest that non-steroidal anti-inflammatory drugs could enhance the antiviral activity of interferon-alpha in chronic HCV infection. In this study, we investigated the effect of indomethacin, a non-steroidal anti inflammatory drug, and interferon-alpha on cytokine production by peripheral blood mononuclear cells from 12 untreated patients with chronic hepatitis C. We evaluated the effect of incubation with indomethacin, interferon-alpha or both on synthesis of Th1- (interleukin-2, interferon-gamma) and Th2-associated cytokines (interleukin-4, interleukin-10), and of the antiviral protein 2',5' oligoadenylate synthetase. Interferon-alpha induced a significant increase in production of interleukin-2. Smaller increases were also seen in the presence of indomethacin, while incubation with both indomethacin and interferon-alpha leads to a synergistic effect. Incubation with indomethacin decreased both interleukin 4 and interleukin-10, whereas interferon-alpha increased these cytokines. The addition of indomethacin to interferon-alpha significantly reversed this interferon-induced increase. Finally, both indomethacin and the association interferon-alpha plus indomethacin determined a significant increase in 2',5' oligoadenylate synthetase production compared to both baseline and interferon alpha alone. In conclusion, indomethacin was able to enhance the antiviral activity of interferon-alpha and to modulate the interferon-induced Th1 and Th2 cytokine response by increasing the Th1-response, fundamental for sustained clearance of HCV, and by decreasing the Th-2 type response, associated with HCV persistence. PMID- 15135803 TI - Progesterone, but not 17beta-estradiol, increases TNF-alpha secretion in U937 monocytes. AB - The Women Health Initiative Clinical trial results suggest that post-menopausal women receiving estrogen + progesterone are at risk for heart disease compared with estrogen alone supplemented women. We examined the hypothesis that progesterone but not 17beta-estradiol (E) increases the secretion of pro inflammatory cytokines interleukin (IL)-6 and tumor necrosis factor (TNF)-alpha. U937 human monocytes were cultured with normal or high glucose in the presence and absence of estrogen or progesterone at 37 degrees C for 24 h. Results show that estrogen inhibits IL-6 but not TNF-alpha secretion (p < 0.05) in monocytes activated by lipopolysaccharide (LPS) or high glucose. In addition, progesterone increased the TNF-alpha secretion in activated monocytes. Thus, progesterone supplementation along with estrogen may increase blood levels of pro-inflammatory cytokine TNF-alpha and thus risk of heart disease in post-menopausal women. PMID- 15135804 TI - Activated protein C inhibits the release of macrophage inflammatory protein-1 alpha from THP-1 cells and from human monocytes. AB - Several lines of evidence have implicated activated protein C (APC) to be an endogenous inhibitor of the inflammatory septic cascade. APC may exhibit direct anti-inflammatory properties, independent of its antithrombotic effects. Chemokines influence the interaction of monocytes at the endothelium during infection and sepsis and are involved in the molecular events leading to an adverse and lethal outcome of sepsis. Defining regulatory mechanisms on the monocytic release profile of the proinflammatory C-C chemokines macrophage inflammatory protein-1-alpha (MIP-1-alpha) and monocyte chemoattractant protein-1 (MCP-1) might have therapeutic implications for the treatment of sepsis. We established a monocytic cell model of inflammation by the addition of lipopolysaccharide (LPS) and examined the effect of human APC on LPS-stimulated chemokine release from the monocytic cell line THP-1. We found that human APC in supra-physiological concentrations of 2.5-10 microg/ml inhibited the LPS-induced release of the chemokines MIP-1-alpha and MCP-1, as measured by enzyme-linked immunosorbent assays (ELISA) at 6 up to 24 h. In addition to experiments on THP-1 cells, recombinant human APC in concentrations of 50 ng/ml was found to have an inhibiting effect on the release of MIP-1-alpha from freshly isolated mononuclear cells of septic patients. The ability of APC to decrease the release of the C-C chemokine MIP-1-alpha from the monocytic cell line THP-1 and from human monocytes may identify a novel immunomodulatory pathway by which APC exerts its anti inflammatory action and may contribute to control the inflammatory response in sepsis. PMID- 15135805 TI - Polymorphisms of chemokine and chemokine receptor genes in Type 1 diabetes mellitus and its complications. AB - Cytokines and chemokines have been implicated in the pathogenesis of Type 1 diabetes mellitus (T1DM) and its microvascular complications. Recently, genetic variants of monocyte chemotactic protein-1 (MCP-1), CC-chemokine receptor 2 (CCR2), CC-chemokine receptor 5 (CCR5) genes have been identified. The aim was to investigate whether genetic variants of the MCP-1 G(-2518)A, CCR2B 64I, CCR5 G(59029)A, and CCR5 Delta32 are associated with T1DM and its microvascular complications. Two hundred and sixty patients with T1DM with and without diabetic microvascular complications, and 104 normal controls were recruited for this study. Genotypes of the MCP-1 G(-2518)A, CCR2B 64I, CCR5 G(59029)A, and CCR5 delta32 were performed by polymerase chain reaction followed by digestion with appropriate restriction endonucleases. Frequencies of the MCP-1 A(-2518) allele (74.6% vs. 63.5%, p < 0.003) and A/A genotype (54.5% vs. 34.6%, p < 0.001, Pc = 0.002) were significantly higher in patients with T1DM compared with normal controls. CCR5 G(59029) was slightly increased in the patients with microvascular complications compared with the uncomplicated (21.4% vs. 10%, p < 0.03, Pc = ns). The frequency of haplotype G/G/W was slightly increased in the patients with diabetic complications compared to the uncomplicated (39.6% vs. 28.8%, p < 0.02, Pc = ns). These results suggest that polymorphisms of the MCP-1, CCR2 and CCR5 genes may be associated with T1DM and its complications. PMID- 15135806 TI - IL-9 increases the expression of several cytokines in activated mast cells, while the IL-9-induced IL-9 production is inhibited in mast cells of histamine-free transgenic mice. AB - Histamine and IL-9 are suspected to play an important role in the pathogenesis of asthmatic and allergic reactions. Mast cells store a large amount of histamine in their granules and are capable of producing different cytokines upon stimulation. In this study we show that mast cells stimulated by IL-9 and ionomycin or IL-9 and antigen-specific IgE/antigen express several cytokines at mRNA level, among them are IL-5, IL-4, IL-10, IL-9, IL-13, IL-1beta, IL-1Ra, IL-6 and MIF. Furthermore, both IL-9 and ionomycin are needed for the production of these cytokines in great quantities, which is mediated through the production of IL 1beta. Histamine-free mast cells respond by a markedly decreased IL-9 expression to this stimulation. Our results show that this IL-9-induced IL-9 production may result in a positive feedback loop in mast cells and the lack of histamine disturbs this loop, which may serve as an explanation for the reduced asthmatic symptoms, observed in histamine-free mice. PMID- 15135807 TI - Increased Th1 activity in patients with coronary artery disease. AB - BACKGROUND: Atherosclerotic lesions are mainly composed of macrophages and T lymphocytes. Specific T helper type 1 (Th1) cytokines and interferon gamma (IFN gamma) inducible chemokines have been shown to be present in these lesions, modulating the local immunologic response. To explore whether this increase in Th1 activity could also be detected in circulating cells indicating a systemic activation, we studied the peripheral expression of Th1 cytokines and chemokines in patients with coronary artery disease and controls. METHODS AND RESULTS: Fifty patients with coronary artery disease (25 with unstable angina and 25 with stable angina) and 10 controls were studied. Serum interleukin (IL)-12 and IFN-gamma and the expression of IFN-gamma inducible chemokines IP-10, Mig and their receptor CXCR3 in peripheral cells were analyzed. Serum IL-12 and intracellular expression of IFN-gamma were significantly elevated in patients with unstable angina. An enhanced expression of IFN-gamma chemokines IP-10, Mig and CXCR3 in patients with stable angina was also observed. CONCLUSIONS: This study demonstrates an increased systemic inflammatory activity in patients with coronary heart disease with a predominant Th1 response, particularly in patients with unstable angina, suggesting an important role played by this polarization in plaque formation and rupture. PMID- 15135809 TI - Comprehensive study on the epidemiological, physiological, and pathological aspects of sudden infant death syndrome. PMID- 15135808 TI - Human interleukin-1-induced murine osteoclastogenesis is dependent on RANKL, but independent of TNF-alpha. AB - Although interleukin-1 (IL-1) has been implicated in the pathogenesis of inflammatory osteolysis, the means by which it recruits osteoclasts and promotes bone destruction are largely unknown. Recently, a cytokine-driven, stromal cell free mouse osteoclastogenesis model was established. A combination of macrophage colony stimulating factor (M-CSF) and receptor activator of NFkappaB ligand (RANKL) was proven to be sufficient in inducing differentiation of bone marrow hematopoietic precursor cells to bone-resorbing osteoclasts in the absence of stromal cells or osteoblasts. This study utilizes this model to examine the impact of human IL-1beta on in vitro osteoclastogenesis of bone marrow progenitor cells. We found that osteoclast precursor cells failed to undergo osteoclastogenesis when treated with IL-1 alone. In contrast, IL-1 dramatically up-regulated osteoclastogenesis by 2.5- to 4-folds in the presence of RANKL and M CSF. The effect can be significantly blocked by IL-1 receptor antagonist (p < 0.01). Tumor necrosis factor-alpha (TNF-alpha) was undetectable in the culture medium of differentiating osteoclasts induced by IL-1. Adding exogenous TNF-alpha neutralizing antibody had no influence on the IL-1-induced effect as well. These results show that in the absence of stromal cells, IL-1 exacerbates osteoclastogenesis by cooperating with RANKL and M-CSF, while TNF-alpha is not involved in this IL-1-stimulated osteoclast differentiation pathway. PMID- 15135831 TI - Commonalities in the classical, collapsibility and counterfactual concepts of confounding. AB - OBJECTIVES: Three definitions of confounding are available in the epidemiologic literature, namely, the classical, collapsibility, and counterfactual. The classical and collapsibility definitions are intuitively appealing but, especially in the case of the latter, there are shortcomings. The more recent counterfactual definition overcomes these limitations but at the cost of increased abstraction. One of the aims of this article is to demonstrate that under certain conditions the three definitions of confounding have key features in common. CONCLUSIONS: The counterfactual definition of confounding addresses the inherent shortcomings of the classical and collapsibility definitions, and forms the basis of innovative methods of data analysis. PMID- 15135832 TI - Measuring avoidable risks of cancer. PMID- 15135834 TI - The Gini coefficient as a measure for understanding accrual inequalities in multicenter clinical studies. AB - OBJECTIVE: Clinical sites participating in multicenter trials may have unequal performance in recruiting subjects. We propose using the Gini coefficient as a quantitative measure of site accrual inequalities. STUDY DESIGN AND SETTING: We evaluated the relationship of this metric to other study characteristics across 166 clinical studies (27,865 subjects) conducted by the AIDS Clinical Trials Group between 1986 and 1999. RESULTS: Overall there was a modest recruitment inequality among clinical centers (mean Gini=0.33). In multivariate modeling, site accrual inequalities were higher when there was more protracted enrollment, and a larger number of sites and were lower in antiretroviral studies than other studies. In long-term studies, the site accrual inequality increased significantly over time (P=0.004). In efficacy trials, a higher Gini coefficient was associated with higher likelihood of the study results being statistically significant (P=0.010). CONCLUSION: The Gini coefficient may be easily and routinely incorporated in the description of the characteristics of a clinical study and may provide insights about its enrollment pattern. PMID- 15135833 TI - Validation of the Harvard Cancer Risk Index: a prediction tool for individual cancer risk. AB - OBJECTIVE: Risk appraisal tools are increasingly being used in the clinical setting to estimate individuals' risks of developing and dying from diseases. The Harvard Cancer Risk Index is one such tool constructed to predict the risks of individuals, aged 40 and above, for developing the leading types of cancer in U.S. men and women relative to the general population. To date, the Risk Index has not been prospectively validated. STUDY DESIGN AND SETTING: Over a period of 10 years' follow-up in the Nurses' Health Study and the Health Professionals' Follow-up Study, age-standardized incidence ratios for cancer of the ovary, colon, and pancreas were calculated for the Risk Index's relative risk categories to assess goodness of fit for risk prediction at the aggregate level. Age adjusted concordance statistics were determined as measures of discriminatory accuracy at the individual level. RESULTS: The Risk Index was well calibrated with observed relative risks across categories for ovarian and colon cancer in women and pancreatic cancer in men, while it performed moderately for colon cancer in men. Discriminatory accuracy was modest for ovarian cancer (age adjusted concordance statistic = 0.59), and relatively good for pancreatic cancer (concordance statistic of 0.72), and colon cancer in men and women (concordance statistics of 0.71, 0.67 respectively). CONCLUSION: The results of this prospective validation provide evidence for the validity of the Risk Index in predicting individuals' risks of cancers, and thereby offer support for future applications of this risk appraisal tool. PMID- 15135835 TI - Capture-recapture is a potentially useful method for assessing publication bias. AB - OBJECTIVES: Publication bias is a problem in systematic reviews of randomized controlled trials. The aim of this study was to assess the influence of publication bias in a systematic review of the effectiveness of Progressive Resistance Training (PRT) in older people. STUDY DESIGN AND SETTING: The relevant studies were ascertained from three sources: electronic databases, experts, and handsearching. Capture-recapture, visual inspection of funnel plots, two statistical tests, and two methods that make adjustments for publication bias, were employed to check the robustness of the conclusions of the systematic review. RESULTS: The methods employed gave broadly consistent results. Capture recapture estimated that 3 (95% CI ) relevant studies were missed, while Trim and Fill suggested 16 studies had been missed. Both Egger's test for bias and a funnel plot regression approach suggested that publication bias was present. A selection model approach suggested that the funnel plot asymmetry observed may not be entirely due to publication bias. CONCLUSION: Capture-recapture is a potentially useful method for assessing publication bias. Further research in the form of simulation studies is required, using a variety of scenarios to investigate the extent to which each method approximates the truth. PMID- 15135836 TI - A statistical simulation study finds discordance between WHO criteria and RECIST guideline. AB - OBJECTIVE: Tumor shrinkage has been adopted as an end point for evaluating the effectiveness of new anticancer agents. The WHO (World Health Organization) criterion suggested measuring the tumor shrinkage by the change in the product of maximal diameter (MD) and the corresponding largest perpendicular diameter (LPD). The RECIST (Response Evaluation Criteria In Solid Tumor) guideline proposed using the change in MD only, based on the observation that this measure is more linearly related to tumor cell kill than the cross product (MD*LPD). Both criteria classify patients into four categories of response: complete response (CR: total disappearance), partial response (PR), stable disease (SD), and progressive disease (PD) but the criteria used in the definition of PD vary. It was anticipated that patients' actual response categorization would not be considerably affected by utilizing the RECIST criteria instead of WHO. Empirical evidence supporting this fact was provided by retrospective analysis of several large datasets. STUDY DESIGN AND SETTING: A statistical simulation is performed to generate tumor measurements and patient response data under meaningful probability distributions with parameters based on data from 130 patients on clinical trials at a cancer center. Concordance measures between the two response criteria (Kappa coefficient and percentage disagreement per response category) are assessed systematically over various combinations of the percentage of elliptical tumors at baseline and the percentage of tumors changing shape from baseline to follow-up. RESULTS: The overall percentage of disagreement between the two methods of response assessment is found to be in the range of 14-20%. The patients categorized by WHO in the PR, SD, and PD groups fall into a different category when assessed by RECIST between 8-16%, 3-12%, and 32-35% of the times, respectively. The kappa coefficient ranges between 0.68-0.77. The proportion of elliptical tumors at baseline does not greatly impact the concordance, but the magnitude of the change in the aspect ratio has a large impact. CONCLUSION: Response assessment as measured by RECIST, with both a change in the underlying metric and change in definition of progression, often results in different categorization of response compared to WHO. The difference in response categorization may be problematic when new experimental therapies are compared to conventional agents whose response rates have been established in historical trials. The apparent lower rate of disease progression with RECIST may mean that more patients remain on therapy. Higher percentages of patients with SD need to be interpreted cautiously by distinguishing those due to the change in the response criterion as opposed to those induced by drugs using pathways such as angiogenesis where disease stabilization is expected rather than shrinkage of tumor. PMID- 15135837 TI - Assessing accuracy of diagnosis-type indicators for flagging complications in administrative data. AB - OBJECTIVE: Canadian administrative hospital discharge data contain a diagnosis type indicator for each coded diagnosis that allows researchers to distinguish complications from pre-existing diagnoses. Given that the validity of diagnosis type indicators is unknown, we conducted a detailed chart review to evaluate the accuracy of diagnosis-type indicators for flagging complications. STUDY DESIGN AND SETTING: We obtained administrative hospital discharge data for 1,200 randomly selected adult inpatient separations in Calgary, Alberta, occurring between April 1, 1996 and March 31, 1997. Each discharge record contains up to 16 diagnoses and 16 corresponding diagnosis-type indicators (value of "2"=complication). The corresponding medical charts were reviewed for evidence of diagnoses and complications. A complication was defined as a new diagnosis arising after the start of hospitalization. We determined the extent to which the diagnosis-type indicator in the administrative data agreed with the chart reviewer's assessment (criterion standard) of whether a diagnosis was a complication or not. RESULTS: The agreement for complications between the two databases varied greatly across 12 conditions studied (kappa range: 0-0.72) and was often low (kappa <0.20 for six conditions). Sensitivity ranged from 0 to 57.1% (higher than 50% for only two conditions), indicating a tendency for complications to often be miscoded as baseline comorbidities. In contrast, specificity was generally high (range: 99.0-100%), suggesting that pre-existing conditions were usually appropriately coded as such in the administrative data. CONCLUSION: The validity of diagnosis-type indicators in Canadian administrative discharge data appears to be poor for some types of complications. This is likely to be of greatest concern in studies that rely solely on diagnosis-type indicators to define complications as outcomes. PMID- 15135838 TI - Alcoholism treatment episodes validly defined using mental health care utilization records. AB - OBJECTIVE: We propose a method for defining and empirically validating episodes of alcoholism treatment from health care utilization records. STUDY DESIGN AND SETTING: The study includes utilization records from 86,207 patients enrolled in a large managed behavioral care company who had at least one alcoholism encounter between 1991 and 1998. Treatment episodes are defined as a minimum number of alcoholism treatment encounters with the behavioral care company prior to a "clear zone" of no encounters. Statistical procedures to select a subset of episode definitions from a number of candidate definitions and methods for assessing the convergent and criterion validity of the definitions are presented. RESULTS: The percentage of patients having at least one episode of alcoholism treatment varies from 43% to 77%, with the results being more sensitive to the minimum number of encounters required than the length of the clear zone. Criterion validity does not reveal any clear "winning" definitions; positive predictive ability increases most rapidly when going from 2 to 3 encounters required. CONCLUSION: The most robust definitions of an alcoholism treatment episode entail 3 to 4 encounters with a clear zone of 3 to 4 months. PMID- 15135839 TI - Psychosomatic status affects the relationship between subjective hearing difficulties and the results of audiometry. AB - OBJECTIVE: Subjective hearing difficulties are often used as a surrogate to audiometry in health check-up, although its effectiveness has not been tested in healthy workers. We conducted a study to test the usefulness of self-reported hearing difficulties for screening hearing impairment among healthy workers by comparing the results with those of audiometry. STUDY DESIGN AND SETTING: This was a cross-sectional observational study. A sample of 12,495 healthy workers in Japan without excess noise exposure was recruited at regular health check-up. Audiometry was conducted after self-administered questionnaire asking symptoms including hearing difficulties. The results were compared to those of audiometry as the gold standard. RESULTS: Overall prevalence of hearing impairment was 4.6% among the population. Subjective hearing difficulties showed 0.21 of sensitivity, 0.95 of specificity, and 0.93 of accordance against audiometry. When limited to those with more than two psychosomatic symptoms, however, the corresponding numbers were 0.43, 0.85, and 0.83. CONCLUSION: The assessment of subjective hearing difficulty may not be able to replace audiometry, but may be useful to detect psychosocial problems of hearing in the workplace. PMID- 15135840 TI - A case-control study found that low albumin and smoking were associated with aortic dissection. AB - OBJECTIVE: Hypertension is a risk factor for aortic dissection. Besides hypertension, associated factors for incidence of aortic dissection are not clear. The aim of this study was to examine whether other factors exist by an epidemiologic case-control approach. STUDY DESIGN AND SETTING: 240 consecutive cases suffering from acute aortic dissection were hospitalized in our university from 1989 to 2000. Blood chemistries and other information obtained immediately after admissions for the first attack were available in 226 patients. A case control approach was performed by frequency matching age, sex, and history of hypertension in the remaining 226 cases with those of 226 healthy controls drawn from a similar geographic catchment area and all observed in 1989. RESULTS: The mean albumin (3.6 +/- 0.5 vs. 4.3 +/- 0.2 g/dL: P<.001) level of cases was significantly lower than that of controls (odds ratio: 0.004 per g/dL). The frequency with which cases of current smokers was significantly higher than that of controls (odds ratio: 3.475). The above parameters of cases compared with controls were statistically significant after adjustments for age, sex, and history of hypertension. CONCLUSION: Low albumin levels and smoking in addition to hypertension are significantly associated with aortic dissection. PMID- 15135841 TI - Searching multiple clinical information systems for longer time periods found more prevalent cases of asthma. AB - OBJECTIVE: The development of a reliable asthma registry is an important first step for conducting population-based asthma disease management. This study developed a computerized algorithm for defining prevalent asthma, identified operational difficulties, and summarized data on asthma prevalence in the study population. STUDY DESIGN AND SETTING: As part of a study of the incidence of occupational asthma, we used the electronic databases of a large health maintenance organization to develop a computerized algorithm for defining prevalent asthma and validated it against chart review. The predictive values of eight health care utilization profiles were validated by chart review to establish the algorithm. RESULTS: The 1-year treated prevalence of asthma was 4.1% among members aged 15-55; the pharmacy database identified 61% of cases, and the outpatient care database 66%. Extending the outpatient care window from 1 year to 2 years increased estimated prevalence to 5.3%, with 81% now found in the outpatient care database. CONCLUSION: This analysis illustrates the benefit of using multiple databases for more accurate enumeration of cases and the impact of extending the search in time. These results are useful for researchers who can use such databases in selecting algorithms to define and identify asthma for their own purposes. PMID- 15135842 TI - Do women with tuberculosis have a lower likelihood of getting diagnosed? Prevalence and case detection of sputum smear positive pulmonary TB, a population based study from Vietnam. AB - OBJECTIVE: The aim was to estimate the gender-specific prevalence of tuberculosis (TB) through screening. A further aim was to calculate case detection within the Vietnamese National TB program. STUDY DESIGN AND SETTING: A population-based survey of 35,832 adults was performed within an existing sociodemographic longitudinal study in Bavi district, northern Vietnam. Cases were identified by a screening question about prolonged cough and further diagnosed with sputum examination and a chest X-ray. RESULTS: The estimated prevalence of pulmonary TB among men was 90/100,000 (95% CI 45-135/100,000) and among women 110/100,000 (95% CI 63-157/100,000). Case detection in the district was estimated to 39% (95% CI 20-76%) among men and 12% (95% CI 6-26%) among women. CONCLUSION: TB prevalence was similar among men and women. Case detection among men and women was significantly lower than the reported national case detection of 80%, and there was a significant underdetection of female cases. These findings warrant actions, and emphasize the need to perform similar studies in different contexts. PMID- 15135843 TI - An adaptation of Charlson comorbidity index predicted subsequent mortality in a health survey. AB - OBJECTIVE: The Catalan Health Interview Survey Follow-up Study analyzed survival differences according to comorbidity, using an adaptation of the Charlson's index. STUDY DESIGN AND SETTING: Vital status was ascertained by record linkage with death certificates 5 years after interview. Three thousand one hundred five men and 3,536 women aged 40-84 years old were included in the analysis. Proportional hazards models with age as time scale were used to calculate relative risk (RR) and 95% confidence interval adjusted for potential confounders. RESULTS: The adjusted RR of death in men was 1.02 (0.73-1.41) for a comorbidity index of 1-2; the RR was 1.51 (1-2.30) for an index of 3-4, and 2.64 (1.43-4.89) for an index of >4 composed to an index of 0. In women, for the same comorbidity index categorization, the RR of death were 0.83 (0.55-1.24), 1.71 (1.09-2.72) and 2.65 (1.47-4.77). CONCLUSION: This result confirms the relation between comorbidity and the risk of death based on a comorbidity index that takes into account severity and number of self-declared chronic diseases with mortality. PMID- 15135844 TI - Positive life orientation as a predictor of 10-year outcome in an aged population. AB - OBJECTIVE: It is well known that depression predicts mortality in old age. However, little is known about the impact of positive emotions. We investigated the impact of positive life orientation on mortality and permanent institutional care in aged birth cohorts. STUDY DESIGN AND SETTING: Participants (born 1904, 1909, and 1914) underwent detailed assessments with follow-up at 5 and at 10 years. Positive life orientation was determined as answering "yes" to all the following items: being satisfied with life, having zest for life, having plans for the future, feeling needed, seldom feeling lonely or depressed. RESULTS: Of participants, 102 (20.8%) had a positive life orientation. After 10 years, 54.5% of them were alive, whereas in the rest of the sample 39.5% survived (P=.004). After controlling for age, gender, and health measures, the impact of positive life orientation was still significant (HR=0.89, 95% CI 0.83-0.93). At 5 years, only 2.9% of those having a positive life orientation but 17.5% of the rest of the sample were in permanent institutional care (P=0.003), with a positive life orientation remaining a significant protector against institutional care (OR 0.58, 95% CI 0.36-0.93). CONCLUSION: Positive attitudes have a long-standing impact on prognosis in old age. PMID- 15135845 TI - A follow-up study found that cardiovascular risk in middle age predicted mortality and quality of life in old age. AB - OBJECTIVE: Cardiovascular risk reduction, while saving lives, may prolong the time with disability and impair the quality of life in survivors. We compared the consequences of middle age cardiovascular risk in old age. STUDY DESIGN AND SETTING: In 1974, risk was low in 593 (low-risk group) and high in 610 men (high risk group). At baseline, all were healthy with similar age and socioeconomic status. Lifestyle and clinical factors, including quality of life (RAND-36), were surveyed with a questionnaire in 2000, and mortality was determined up to 2002. RESULTS: During the follow-up, 303 men died, with mortality 54% higher in the high-risk group (P=.001). In the 2000 survey, high-risk men still had significantly greater BMI, higher blood glucose, higher prevalence of smoking, and more sedentary lifestyle, and they reported more both cardiovascular and noncardiovascular diseases. All the RAND-36 scales were worse in the high-risk group; of the two component summary scores, physical (PCS), but not mental (MCS) score, was significantly lower in the high-risk group. CONCLUSION: Low cardiovascular risk in middle age was associated with lower mortality, morbidity, and better quality of life in old age 26 years later. The results may support the theory of compression of morbidity. PMID- 15135846 TI - A case-control study on adverse effects: H2 blocker or proton pump inhibitor use and risk of vitamin B12 deficiency in older adults. AB - OBJECTIVE: Acid-suppressant drugs are commonly prescribed for elderly patients, a population in which vitamin B(12) deficiency is a common disorder. The purpose of this study was to examine the possible association between use of prescription histamine H-2 receptor antagonists (H2RA) or proton pump inhibitors (PPI) and vitamin B(12) deficiency in older adults. STUDY DESIGN AND SETTING: This was a case-control study in a University-based geriatric primary care setting. Among patients aged 65 years or older with documented serum vitamin B(12) studies between 1990 and 1997, 53 vitamin B(12)-deficient cases were compared with 212 controls for past or current use of prescription H2RA/PPI according to information in subjects' medical records. RESULTS: Controlling for age, gender, multivitamin use, and Helicobacter pylori infection, chronic (#10878;12 months) current use of H2RA/PPI was associated with a significantly increased risk of vitamin B(12) deficiency (OR 4.45; 95% CI 1.47-13.34). No association was found between past or short-term current use of H2RA/PPI and vitamin B(12) deficiency. CONCLUSION: These findings support an association between chronic use of H2RA/PPI by older adults and development of vitamin B(12) deficiency. Additional studies are needed to confirm these findings. PMID- 15135847 TI - Back-up antibiotic prescriptions could reduce unnecessary antibiotic use in rhinosinusitis. AB - OBJECTIVES: To examine the attitudes of patients with rhinosinusitis toward the availability of "back-up" antibiotics, and potential implications for antibiotic use rates. STUDY DESIGN AND SETTING: A survey that assessed actual and hypothetical antibiotic prescription patterns was administered to a convenience sample of patients treated for rhinosinusitis in one acute care facility between September 1 and December 1, 2001. RESULTS: Of 386 eligible patients, 114 completed the survey. Seventy-six percent of patients expected antibiotic treatment; satisfaction rates were significantly associated with receiving an antibiotic prescription (P <.05). Over two-thirds of patients (69.7%) reported preference for back-up antibiotic prescriptions in the future, with 91.1% stating they would wait at least 1 day, and 52.7% at least 7 days, to fill a backup prescription. In sensitivity analysis, back-up prescriptions significantly reduced antibiotic use over a wide range of assumptions. CONCLUSIONS: The majority of patients with rhinosinusitis in this study expected antibiotic prescriptions, and were more satisfied if they were received. Back-up antibiotics have the potential to reduce unnecessary antibiotic use, mitigate risk of nontreatment, and preserve high levels of patient satisfaction. PMID- 15135848 TI - Effect of perinatal (prenatal?) loratadine exposure on male rat reproductive organ development. PMID- 15135850 TI - NTP-CERHR Expert Panel report on the reproductive and developmental toxicity of ethylene glycol. PMID- 15135851 TI - NTP-CERHR Expert Panel report on the reproductive and developmental toxicity of propylene glycol. PMID- 15135852 TI - Comparative studies on the spermatotoxic effects of dinoseb and its structurally related chemicals. AB - Three dinitrophenolic compounds, dinoseb (DNBP; 7.5 mg/kg b.w.), 4,6-dinitro-o cresol (DNOC; 4, 7.5, 15 mg/kg b.w.), and 2,4-dinitrophenol (DNP; 7.5, 15, 30 mg/kg b.w.) were administered orally to sexually matured Jcl:SD male rats for 5 consecutive days. Half of the males in each group were necropsied at 3 (D3) and 14 (D14) days after the last dosing, respectively, and examined for the effects of dinitrophenols on spermato-/spermiogenesis. DNBP (7.5 mg/kg), DNOC (15 mg/kg), and DNP (30 mg/kg) caused 1, 5, and 0 deaths, respectively, as well as a decreased body weights during the treatment. Although examinations on D3 revealed no treatment-related alterations, DNBP and DNOC resulted in reduced sperm motility and increased incidence of tailless sperm in the cauda epididymis on D14. DNP also caused slightly increased incidence of tailless sperm on D14. These results demonstrate that DNBP, DNOC, and DNP manifest similar spermatotoxic effects at or around a lethal dose in rats. PMID- 15135853 TI - Flutamide induces ultrastructural changes in spermatids and the ectoplasmic specialization between the Sertoli cell and spermatids in mouse testes. AB - Flutamide (Flu) is an anti-androgenic compound that disrupts development of male androgen-dependent tissues. The purpose of the present study was to investigate the effects of Flu on ICR mouse testes by electron microscopic observation. Newborn mice were subcutaneously injected with 0.00012, 0.0012, 0.012, 0.12, 1.2, 12 or 120 microg Flu/g body weight/shot on Days 2, 4, 6, 8, 10 and 12 (Day 1: day of birth). In addition, adult mice were injected with 0.0012, 0.012, 0.12 or 1.2 microg Flu/g body weight/day for 5 sequential days. Testes were processed for electron microscopy. In neonatal treatments, acrosomes and/or nuclei of the spermatids were deformed. In addition, the ectoplasmic specialization between the Sertoli cell and spermatids was partially or completely deleted. Stages of the seminiferous cycle were also disarranged in the neonatal treatments. There were no ultrastructural differences between the effects of neonatal and adult treatments, however, stage disarrangement was not observed in adult treatments. The percentages of abnormal spermatids were higher in neonatally treated mice than in mice treated as adults. Since similar observations were reported after treatment with beta-estradiol-3-benzoate (E2B), the presence of Flu may induce a "xenoestrogenic environment" in mouse testes. PMID- 15135854 TI - Temporary impairment of reproduction in freshwater teleost exposed to nonylphenol. AB - New born guppies, Poecilia reticulata, were exposed to a sublethal concentration of nonylphenol (NP) for 90 days, with the aim of evaluating the influence that this pollutant has on reproductive functions. At the end of treatment, the gender balance was significantly biased towards females (sex ratio = 0.3 males per female). Treated males displayed anomalous transcription of vitellogenin in correlation with a significant reduction of the gonadosomatic index (GSI); however, an increase of vitellogenin gene transcription, correlating directly with an increase in GSI, was observed in NP treated females. The hepatosomatic index (HSI) was significantly increased in exposed males and females. Females exposed to NP showed no alteration of sexual behavior or reproduction. These findings in guppies suggest that NP has estrogenic potency sufficient to disturb reproduction. Effects were transitory in nature because three months in uncontaminated water allowed males to recover normal sexual efficiency. PMID- 15135855 TI - Reproductive toxicity assessment of chronic dietary exposure to soy isoflavones in male rats. AB - Epidemiologic and experimental data suggest that consumption of diets that are rich in isoflavones may decrease cancer risk in the breast, prostate, and other tissues. Isoflavones such as genistein and daidzein are structurally similar to endogenous estrogens, and demonstrate both estrogenic and weak anti-estrogenic activities; these activities may underlie the impaired fertility and reproductive tract disorders reported in animals exposed to high doses of isoflavones. To identify possible effects of isoflavones on male fertility, we evaluated reproductive parameters in Wistar-Unilever rats receiving dietary exposure to PTI G-2535, a characterized mixture of soy-derived isoflavones containing 45% genistein, 23% daidzein, and 4% glycitein. Beginning at 10 weeks of age, rats received chronic dietary exposure to the soy isoflavone mixture (200 or 2000 mg/kg diet) for a minimum of 12 months. Controls received unsupplemented chow diet only for the same period. Dietary exposure to isoflavones induced no gross toxicity or alterations in body weight gain. Absolute and relative weights of the testis and epididymis in groups receiving high or low doses of isoflavones were comparable to those of controls, and histopathologic evaluations demonstrated that testicular morphology was similar in all study groups. Isoflavone exposure had no significant effects on spermatid count, sperm production, or sperm morphology in any group. These data suggest that the reproductive system of adult male rats is relatively insensitive to isoflavone toxicity at dose levels that demonstrate significant activity in cancer chemoprevention, and that male reproductive function is unlikely to be affected by long-term administration of isoflavones for cancer prevention or other purposes. The results of this study conducted in adult male rats differ from the significant alterations in reproductive parameters that have been reported in female rats receiving prenatal or juvenile exposure to isoflavones. PMID- 15135856 TI - Elimination kinetics of ethanol in pregnant women. AB - To evaluate the pharmacokinetics of ethanol in the early second trimester of pregnancy, ethanol concentrations simultaneously measured in the maternal blood (EtOH-MB) and in the amniotic fluid (EtOH-AF) of six pregnant women were obtained from a previous study in which a single ethanol dose of 300mgkg(-1) body weight was administered orally. For maternal blood ethanol concentration, the kinetic equation was: [Formula: see text] where k(12) and k(21) are, respectively, the rate constant of ethanol transfer from either the central compartment to the peripheral compartment or vice versa; V(max)(EtOH)is the maximal velocity for ethanol oxidation; and K(m)(EtOH) is the concentration at which half of the maximal rate of ethanol elimination is reached. The maximum concentration of EtOH in AF was 60% lower than in MB (P=0.36). However, the AUC(0-3.5h) in AF was only 16% lower than the value for MB (P=0.059). The k(12) (0.20 =/- 0.26 h(-1)) was almost twice faster than k(21) (0, h(-1)). The V(max)(EtOH) was 237.6 +/- 71.5 microgml(-1)h(-1) and K(m)(EtOH) was 3.7 +/- 4.7 microgml(-1). Our results imply that in the early second trimester, ethanol metabolism is fast. However, ethanol clearance from the AF is slower than ethanol clearance in MB. This process is widely variable, and our findings may partially explain the wide variability of ethanol's toxic effects on the fetus. PMID- 15135857 TI - Normal pregnancy outcome following inadvertent exposure to rosiglitazone, gliclazide, and atorvastatin in a diabetic and hypertensive woman. AB - The subject is a diabetic and hypertensive woman treated early during an unplanned pregnancy with a multi-drug regimen that included three drugs with no prior history for use in pregnant women (rosiglitazone, gliclazide, atorvastatin). She was under care for chronic hypertension, which she suffered for 14 years, and diabetes mellitus, hypercholesterolemia, anxiety disorder, morbid obesity and epilepsia for 5 years. She was exposed to rosiglitazone (4mg/day), gliclazide (60mg/day), and atorvastatin (40mg/day) in addition to acarbose, spironolactone, hydrochlorothiazide, carbamazepine, thioridazine, amitryptiline, chlordiazepoxide, and pipenzolate bromide during the first 7 weeks of gestation while unaware of pregnancy. Pharmacotherapy was adjusted following clinical recognition of pregnancy during the 8th week. She gave birth to a normal healthy infant at the 36th week of gestation. This is the first reported case of human exposure to rosiglitazone, gliclazide, and atorvastatin during pregnancy. Although the normal pregnancy outcome does not address the safety of these drugs for use in pregnancy, these data contribute to a limited knowledge regarding human exposure to these antidiabetic drugs. PMID- 15135858 TI - Histopathological studies of visceralized Leishmania (Leishmania) amazonensis in mice experimentally infected. AB - BALB/c, C57BL/6, and DBA/2 mice were subcutaneously infected in the left footpad by injecting 10(4) Leishmania (Leishmania) amazonensis amastigotes. Mice were sacrificed 20, 30, 40, 60 and 90 days post-infection. Fragments of liver, kidney, spleen, skin, and draining lymph node were collected for histological examination. Light microscopy showed that at 20 days after infection BALB/c mice presented discrete inflammatory infiltrates in the skin made up of eosinophils, lymphocytes, and rare parasitized macrophages. Ninety days post-infection, the dermis showed necrotic tissue, large numbers of mononuclear cells and vacuolated macrophages filled with amastigotes. Forty days post-infection, the draining lymph nodes showed hyperplastic germinal centers, increase of high endothelial venules and apoptosis in germinal center cells. After the first 3 months post infection, the involvement of spleen, kidney and liver was discreet, being characterized by multifocal inflammatory infiltrates. Eight months after infection, the animals presented metastatic lesions in the contralateral footpad and nose. In deep dermis, there was remarkable proliferation of fibroblasts associated with collagen fibers. The liver showed multifocal granulomas and mononuclear infiltrate around the blood vessels, but no parasites were observed, except in one animal. In some mice there were immature cells of the hematopoietic lineage. Both BALB/c and C57BL/6 mice presented osteonecrosis, which is characterized by pycnotic osteocytes and empty lacunae at the point of inoculation and subsequently, replacement of this tissue by fibrous connective tissue and colonization of the bone marrow. A diffuse inflammatory process composed of mononuclear cells and rare parasites were seen in the kidneys. In one mouse, bone marrow cells were observed in the renal medulla along with where free amastigotes. DBA/2 mice developed a mild infection and they did not visceralize. In conclusion, our data demonstrates that in susceptible mice L. (L.) amazonensis, a causative agent of tegumentary leishmaniasis, causes pathological changes similar to those produced by Leishmania (L.) infantum in both humans and canids. PMID- 15135859 TI - World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines for evaluating the efficacy of anticoccidial drugs in chickens and turkeys. AB - These guidelines have been written to aid in the design, implementation and interpretation of studies for the assessment of drug efficacy against Eimeria species in chickens and turkeys. The information provided deals with many aspects of how to conduct controlled studies in battery cages (dose determination), floor pens (dose confirmation), and commercial facilities (field effectiveness studies), the selection of birds, housing, feeding, preparation of medicated rations, record keeping, diagnostic techniques, and methods for the preparation, maintenance and use of parasites. These guidelines are also intended to assist investigators in conducting specific studies, provide specific information for registration authorities involved in the decision-making process, assist in the approval and registration of new anticoccidial drugs, and facilitate the world wide adoption of standard procedures. PMID- 15135860 TI - Plasma kinetics and efficacy of oral megazol treatment in Trypanosoma brucei brucei-infected sheep. AB - Experimentally infected sheep have been previously developed as an animal model of trypanosomosis. We used this model to test the efficacy of megazol on eleven Trypanosoma brucei brucei-infected sheep. When parasites were found in blood on day 11 post-infection, megazol was orally administered at a single dose of 40 or 80mg/kg. After a transient aparasitaemic period, all animals except two relapsed starting at day 2 post-treatment, which were considerated as cured on day 150 post-treatment and showed no relapse after a follow-up period of 270 days. In order to understand the high failure of megazol treatment to cure animals, a kinetic study was carried out. Plasma concentrations of megazol determined, by reverse-phase high-performance liquid chromatography at 8h post-treatment in these animals, were lowered, suggesting slow megazol absorption, except in cured animals. However, megazol plasma profiles in uninfected sheep after a single oral dose of megazol showed a fast megazol lowered absorption associated with a short plasma half-life of drug. Inter-individual variation of megazol pharmacokinetic properties was also observed. These findings suggested that the high failure rates of megazol treatment were related to poor drug availability after oral administration in sheep. In conclusion, megazol could cure sheep with T. b. brucei infection but oral administration was not an effective route. PMID- 15135861 TI - Determination of an optimized cut-off value for the Neospora agglutination test for serodiagnosis in cattle. AB - A definitive diagnosis of neosporosis in cattle implies the examination of the aborted fetus. However, in many instances fetal material is not available. Therefore, most diagnosis are based on serological tests. At the moment, there are no consensuses about the cut-off for serodiagnosis of neosporosis in cattle, for any test. The objective of the present study was to estimate the best cut-off for the Neospora agglutination test (NAT) for serodiagnosis in cattle. For that purpose, 246 serum samples from 4 groups of dairy cows (aborted Neospora positive; not aborted healthy; aborted other diseases and herds endemic neosporosis) were collected and antibodies anti-N. caninum were determined by NAT. Additionally, immunoblot (IB) was performed with sera from all cows of the endemic neosporosis group and the patterns of seroreactivity were contrasted with the NAT titers for this group of cows. Evaluation of the optimized sensitivity and specificity was calculated using Youden's J-statistics. The best Youden's J statistic was obtained at 1:40 titer, presenting 100% of sensitivity and 90.4% of specificity with negative and positive predictive values of 100 and 75.0%, respectively. The comparison between NAT titers and the IB banding pattern support the results of the statistical analysis, i.e. titers of 1:40 and higher showed a complex pattern of bands, while titers lower than 1:40 did not precipitate any bands. These results indicate that 1:40 was an optimized NAT cut off for serodiagnosis in cattle. PMID- 15135862 TI - The efficacy of some drugs with known antiprotozoal activity against Histomonas meleagridis in chickens. AB - Nine drugs with known or suspected antiprotozoal activity were tested in vitro, and in vivo for activity against Histomonas meleagridis. The nitroimidazoles dimetridazole, metronidazole, ornidazole, and tinidazole suppressed growth of H. meleagridis in vitro at 10 microg/ml or higher. Paromomycin sulfate, and carbadox were weakly effective at high levels. Quinolinol, mebendazole, diloxanide furoate, and albendazole had no demonstrable efficacy in vitro. Drugs showing some activity in vitro were tested in young chickens inoculated intracloacally with 2 x 10(5) H. meleagridis/bird. Dimetridazole, metronidazole, ornidazole, and tinidazole were highly effective at 200 ppm in feed. Paromomycin sulfate, and carbadox were ineffective in vivo, with no improvement in liver or cecal lesion scores compared to that of infected controls. Thus, the only new entities with efficacy against blackhead disease in vivo were nitroimidazoles, related to the positive control dimetridazole. PMID- 15135863 TI - Rapid detection of Leishmania infantum infection in dogs: comparative study using an immunochromatographic dipstick rk39 test and direct agglutination. AB - A rapid, sensitive and specific tool for detection of Leishmania infantum infection in dogs, would be highly desirable, because it would allow control interventions in endemic areas of Zoonotic visceral leishmaniosis (ZVL). In this study, we compared an immunochromatographic dipstick test with direct agglutination test (DAT) for detecting L. infantum infections in dogs from areas of ZVL endemic in Iran. The validity of the dipstick rk39 (Cypress Diagnostic Company, Belgium) for canine visceral leishmaniosis (CVL) was compared with a standard direct agglutination test on 116 clinically suspected dogs and 152 healthy controls from endemic areas of Ardabil and East Azerbaijan provinces, north-western of Iran for 1 year. A sensitivity of 70.9% and specificity of 84.9% were found at a 1:320 cut off titer when DAT confirmed cases were compared with healthy control. As the dipstick rk39 test is rapid, noninvasive and does not require much expertise or elaborate equipment, it can be used for screening and diagnosis of canine visceral leishmaniosis in remote endemic areas. PMID- 15135864 TI - Characterisation of Theileria parva isolates from Kiambu district, Kenya. AB - Four Theileria parva isolates from Muguga area of Kiambu district, Kenya, were used to establish schizont-infected cell lines. Their protein antigens were then separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS page). The isolates were subsequently subjected to protein analysis and characterisation by the western immunoblotting technique. Probing for the polymorphic immunodominant molecule (PIM) was done using monoclonal antibody no. 4. SDS page detected up to 20 protein antigens of molecular mass 35,000-180,000 Da. The western blot analysis revealed a greater heterogeneity in the molecular mass (M(r)) of PIM than previously thought. The M(r) of PIM varied between 80 and 90 kDa. The isolates further revealed different densities of surface epitopes with variable reaction to the monoclonal antibody. The implications of these findings to the epidemiology of east coast fever and immunisation programmes are discussed. PMID- 15135865 TI - Neospora caninum seroprevalence in dairy cattle in central Thailand. AB - The seroprevalence, in dairy cattle, of antibodies to Neospora caninum, the relationship between seropositivity and age (heifer versus cow), the relationship of herd infection with herd size and the relationship of herd infection with the presence of dogs on the farm were studied. The study involved 549 cows and 82 dogs in 59 dairy herds in Nakhon Pathom, Thailand. A competitive enzyme-linked immunosorbent assay (cELISA) with NC-specific monoclonal antibody was used to detect the NC antibodies in the sera. Individual and herd seroprevalence of NC were 5.5% (30/549) and 34% (20/59), respectively. No significant relationships between NC seropositivity with the age of the cows (heifer versus cow; P > 0.05) and between herd infection and the presence of dogs on the farm (P > 0.05) were found. Herd size significantly affected herd infection (P < 0.05) with higher infection in large than small herds (> or = 21 versus < or = 20 cows). Of 12 cows with a history of abortion, one was seropositive to NC. The seroprevalence of NC antibodies in dogs was 1.2% (1/82). This is the first NC seroprevalence study in dogs in Thailand. It was concluded that Neospora infection was more common at the herd level rather than the individual level in Thailand and the presence of dogs on the farm was not related to the level of herd infection. Caution should be taken in the interpretation of serological tests from the farm dogs. PMID- 15135866 TI - Light and scanning electron microscopy of the miracidium of Echinostoma paraensei (Trematoda, Echinostomatidae). AB - Echinostoma paraensei was described 1967 by Lie and Basch. Recently its natural definitive host, the aquatic rodent Nectomys squamipes, endemic in Brazil, was identified. As most of the echinostomatids, this species presents a zoonotic potential. The morphology and topography of the E. paraensei miracidium obtained from adult worms collected from the natural definitive host N. squamipes is described by light and electron microscopy. The arrangement and the dimensions of the epidermal plates are given. The eyespots are composed by two pairs of lenses measuring 6.03 microm. SEM observations shows that the miracidium body is covered by cilia except at the terebratorium region and the presence of alpha-tubulin in the cilia of the larvae is first recorded by immunelabeling. Nineteen papilla like structures arranged in three axes and four groups were observed at the terebratoriun, this structure is retractable, presenting folds and a corrugated surface with profiles of cytoplasmic expansions, forming network of anatomizing folds. PMID- 15135867 TI - Efficacy of abamectin against ivermectin-resistant strain of Trichostrongylus colubriformis in sheep. AB - The efficacy of two formulations of abamectin, i.e. oral and injectable was determined against ivermectin-resistant strain of T. colubriformis in sheep. Twenty-four lambs were infected with 10,000 third stage larvae of ivermectin resistant strain of T. colubriformis. Twenty-four days post-infection, the lambs were divided randomly into four groups of six animals each according to egg counts. The first group was left untreated and kept as a control. The second group was treated with ivermectin (oral) at 0.2mg kg(-1) body weight. The third group was treated with oral formulation of abamectin at 0.2mg kg(-1) body weight. The fourth group was treated with injectable formulation of abamectin at 0.2mg kg(-1) body weight. Fecal egg count and controlled slaughter tests were employed to determine the efficacy of abamectin (oral and injection) against ivermectin resistant strain of T. colubriformis in sheep. Reduction in arithmetic mean fecal egg counts achieved by ivermectin (oral), abamectin (oral) and abamectin (injection) was 66, 98 and 76%, respectively 10 days after treatment. Ivermectin (oral), abamectin (oral) and abamectin (injection) reduced arithmetic mean worm burden by 63, 97 and 74%, respectively. The findings demonstrated that abamectin oral formulation was more effective than abamectin injection against ivermectin resistant strain of T. colubriformis in sheep. PMID- 15135868 TI - Dermal safety study with imidacloprid/moxidectin topical solution in the ivermectin-sensitive collie. AB - A study was conducted to determine the safety of the dermal application of 10% imidacloprid/2.5% moxidectin topical solution in ivermectin-sensitive collies. Each milliliter of this solution contains 100mg of imidacloprid and 25mg of moxidectin. A total of 21 collies were prescreened for ivermectin-sensitivity and heartworm negative status prior to selection for the study. Animals were assigned based on the maximum ivermectin-sensitivity score demonstrated during the prestudy screening. Treatment groups included a 3x and 5x test article group, and a 3x and 5x mineral oil control group. The 3x and 5x doses were administered at three and five times, respectively, the 1x dose based on the animal's body weight. On day 0, 3 of the 21 dogs were treated with dermal applications of a preliminary dose of 3x test article to screen for unexpected signs of toxicity with the remaining 18 dogs being treated with 3x mineral oil to blind for the volume of liquid applied. After no signs of toxicity were observed, these same three dogs were treated with 3x of test article and 2x mineral oil on days 28 and 56. The remaining 18 animals were equally allocated to either a 5x test article group or a 5x control group and were each treated on days 28, 56, and 84. Personnel performing observations were blinded to treatment. Observations were made for clinical signs of ivermectin sensitivity twice daily during non-dosing days. On treatment days, dogs were observed hourly for the first 4h post treatment and at 6, 8, 12, 18 and 24h. Signs of toxicosis were not observed in any of the dogs throughout the observation period. This study demonstrated the safety of imidacloprid/moxidectin, when administered to collies testing positive for ivermectin sensitivity at dosages up to five times the maximum recommended dose. PMID- 15135869 TI - Ectopic location of adult worms and first-stage larvae of Angiostrongylus vasorum in an infected dog. AB - A mongrel dog with angiostrongylosis died suddenly and was submitted to necropsy. Post-mortem findings showed adult worms in ectopic locations. This is the first report of Angiostrongylus vasorum adult worms being found within the pericardial sac and in the lumen of the bladder of an infected dog. A total of 587 A. vasorum first-stage larvae were recovered from different organs and tissues after concentration using Baermann apparatus. PMID- 15135870 TI - Concurrent parasitic infections of sheep: depression of Trichostrongylus colubriformis populations by a subsequent infection with Oestrus ovis. AB - Concurrent infections of sheep with Oestrus ovis and trichostrongyles of the digestive tract are common in the field. Previous results have shown that a previous infection with O. ovis adversely affects worm populations of either Trichostrongylus colubriformis or Haemonchus contortus. However, no information was available to determine the influence of the succession of infections on the expression of interactions between these parasites located in remote anatomical sites. In order to investigate the role of these modulating factors, an experimental study was conducted on four groups of naive sheep, examining the consequences of a delayed infection with O. ovis on a pre-existing population of T. colubriformis. group T was infected four times with 4000 T. colubriformis larvae on days 0, 14, 28 and 42 of experiment; group O received multiple infections with O. ovis first instar larvae on days 42, 49, 56, 70 and 77; sheep from group TO received both infections and animals from group C remained as uninfected controls. Faecal egg counts and eosinophilia were measured weekly throughout the study. At necropsy (day 91), the mucosal cellular responses in the nasal cavities (septum, turbinates, ethmoid and sinus) and in the digestive tract (stomach and small intestine) from all animals were analysed from histological sections. Infection of the digestive tract with nematodes did not modify the biology of Oestrus populations, as measured by the number and weight of larvae. In contrast, infections with O. ovis after T. colubriformis infection was related to significant reductions (P < 0.01) in nematode egg excretion and worm burdens. These changes were associated with significant modifications in populations of mast cells, globule leucocytes and eosinophils in the respiratory and digestive tracts. These results indicate that an antagonistic interaction exists between the populations of O. ovis in the nasal cavities and T. colubriformis in the small intestine but that the order of succession of infections with the two parasites is not a major modulating factor for expression of interactions. They also confirm that parasitic infection in one particular anatomical site induces "at distance" inflammatory reactions of the whole mucosal system. PMID- 15135871 TI - Genetic diversity and molecular phylogeny of Anaplasma marginale isolates from Minas Gerais, Brazil. AB - Anaplasma marginale (Rickettsiales: Anaplasmataceae), a tick-borne pathogen of cattle, is endemic in tropical and subtropical regions of the world, and many isolates of A. marginale may occur in a given geographic area. Phylogenetic relationships have been reported for A. marginale isolates from the US using gene and protein sequences of MSP1a and msp4. These studies demonstrated that msp4 sequences, but not MSP1a DNA or protein sequences, provide phylogeographic information and also that MSP1a sequences are highly heterogeneous among A. marginale populations. However, little information is available on the genetic diversity of A. marginale isolates from other regions of the world. The present study was undertaken to examine genetic variation among 10 isolates of A. marginale obtained from infected cattle in the State of Minas Gerais, Brazil, where A. marginale is endemic. Neighbor-joining analysis of msp4 sequences of Brazilian and New World isolates of A. marginale from Argentina, Mexico and the US provided bootstrap support for a Latin American clade. The sequences of the MSP1a repeats of four Brazilian isolates of A. marginale were compared to sequences of Latin American and US isolates. The MSP1a repeated sequences of Latin American isolates of A. marginale had nine repeat forms, alpha-phi, which have not been reported previously in North American isolates of A. marginale. Furthermore, the repeated forms tau, sigma and mu were only present in the Brazilian isolates. The results demonstrated that the genetic heterogeneity observed among isolates of A. marginale is common in endemic areas, independent of the predominant tick vector and is consistent with previous studies in which msp4 provided phylogeographic information about A. marginale isolates, while MSP1a was found not to be a useful marker for phylogeographic characterization of A. marginale isolates. PMID- 15135872 TI - High prevalence of Toxoplasma gondii in a commercial flock of chickens in Israel, and public health implications of free-range farming. AB - Little is known of the prevalence of Toxoplasma gondii in commercially raised chickens. In the present study, the prevalence of T. gondii in 96 free-range chickens (Gallus domesticus) from a commercial farm in Israel was assessed. Blood, heart, and brain from each chicken were examined for T. gondii infection. Antibodies to T. gondii, assayed with the modified agglutination test (MAT > or = 1:5), were found in 45 of the 96 chickens. Hearts and brains of seropositive (MAT > or = 1:5) chickens were bioassayed in mice. Additionally, hearts and brains of 51 seronegative (MAT < 1:5) chickens were bioassayed in two T. gondii-free cats. T. gondii was isolated from 19 of the 45 (42.2%) seropositive chickens by bioassay in mice. Both the cats fed tissues pooled from seronegative chickens shed T. gondii oocysts. Tachyzoites and tissue cysts of all 21 isolates of T. gondii from chickens were avirulent for mice. Seventeen of the 19 isolates genotyped were found to be type II, and 2 were type III. Understanding of the sources of infection on such farms could be the key to the development of better prevention strategies. PMID- 15135873 TI - Vertical transmission of Neospora caninum in BALB/c mice in both acute and chronic infection. AB - To examine the frequency of congenital infection by Neospora caninum, BALB/c mice were inoculated intraperitoneally with tachyzoites of N. caninum either during pregnancy (Group 1) or 4 weeks or more before pregnancy (Group 2). Further, the mice inoculated during pregnancy were bred at 4 weeks or more after delivery to form Group 3. Congenital transmission was observed in 76% of the neonates of the mice in Group 1 and in 50% of the neonates of the mice in Group 2. Interestingly, congenital transmission was observed in 86% of the neonates from Group 3. These results suggest that chronically-infected BALB/c mice efficiently transmit N. caninum infection to their offspring. PMID- 15135874 TI - Treatment of small lungworm infestation in sheep by using moxidectin. AB - The use of moxidectin (MXD) in the treatment of small lungworm infestation (Cystocaulus ocreatus, Muellerius capillaris, Neostrongylus linearis and Protostronglylus rufescens) in sheep, was evaluated. Twenty-one sheep naturally infested with small lungworms, were divided into three groups (n = 7) and treated as follows: group A with moxidectin 1% injectable solution at a dose rate of 0.2mgkg(-1) bodyweight, group B with moxidectin 0.1% oral drench at a dose rate of 0.2 mgkg(-1) bodyweight and group C being controls. Before treatment, mean faecal larval counts were 30.7, 21.1 and 26.7 lpg in group A, B and C, respectively; 14 days after treatment respective counts were 0.4, 2.3 and 63.0 lpg, (percentage reduction after moxidectin administration >96.0%); 60 days after treatment respective counts were 0.0, 0.0 and 26.4 lpg, (percentage reduction after moxidectin administration 100%). It is concluded that treatment of small lungworm infestation of sheep can be effected by using moxidectin. PMID- 15135875 TI - Seroprevalence of Toxoplasma gondii antibodies from wild canids from Brazil. AB - The prevalence of anti-Toxoplasma gondii antibodies was evaluated by the indirect immunofluorescent-antibody test in serum of 57 wild canids from three different species: Lycalopex gymnocercus, Cerdocyon thous and Dusicyon vetulus from the northeast, southeast and southern regions of Brazil. The prevalence was 35.1%, with 20 of the 57 canids demonstrating antibodies anti-T. gondii at dilutions of 1:16 in 2, 1:32 in 4, 1:64 in 2, 1:128 in 2, 1:256 in 6, 1:512 in 2 and 1:2048 in 2 animals. None of the D. vetulus were positive. Among the L. gymnocercus 11 (91.7%) of the 12 samples were positive and among C. thous 9 (60%) of the 15 had antibodies anti-T. gondii. PMID- 15135876 TI - Molecular survey of Babesia infection in dogs in Okinawa, Japan. AB - A total of 80 free-roaming dogs on Okinawa Island, Japan, were examined for Babesia infection using the polymerase chain reaction (PCR) and sequence analysis. Of 80 samples, 12 were positive in a Babesia genus-specific PCR. Consequent species-specific PCR for B. canis and B. gibsoni revealed that 5 (6.3%) and 7 (8.8%) dogs were infected with B. canis and B. gibsoni, respectively. Sequence analysis of the PCR products revealed that the 18S rRNA gene sequence of B. canis detected from dogs in Okinawa was very close to B. canis vogeli with sequence similarity of 99.94%. PMID- 15135877 TI - Manfred Zimmermann's contributions to pain research and therapy: dedication on the occasion of his 70th birthday. PMID- 15135879 TI - Stimulating music increases motor coordination in patients afflicted with Morbus Parkinson. AB - The present study measured the short-term effect of special stimulating music on motor coordination in Parkinson patients. Eleven patients with a dominant akinetic Parkinson syndrome as well as ten healthy persons (age-matched control group) participated in this study. In the Parkinson group, the measurement of fine motor coordination with the 'Vienna Test System' showed an improvement in two (aiming, line tracking) of the four subtests after listening to the music. The patients improved their performance with the right arm significantly in the subtest aiming-error-time. No statistical differences were found in the other two subtests (steadiness, tapping) in both groups. There was also no improvement in frequency of tapping movement on the power-force-working-plate. Accordingly, music effects more the precision of a movement than the speediness. The measurements on the power-force-working-plate showed a significant improvement in two of five measured parameters: contact time, variability coefficient for total step and impact maximum changed significantly. This study gives evidence that specific music can improve the precision of arm and finger movements. PMID- 15135880 TI - Relationship between hippocampal volume and CA1 neuron loss in brains of humans with and without Alzheimer's disease. AB - Hippocampal volumetry has been proposed to aid in the early diagnosis of Alzheimer's disease (AD) and to monitor progression of the disease. While this is believed to be as a result of hippocampal neuron loss, this association has not been firmly established and loss of other tissue elements may be responsible for the observed atrophy. We investigated the relationship between neuron loss and hippocampal volume in 11 patients with autopsy-confirmed AD and 11 non-demented age-matched controls. Strong correlations were found between neuron number and both hippocampal volume and brain volume demonstrating that volume and neuron content are related in normal subjects and that the relationship is maintained in AD. In AD, neuron number and volume measures significantly decline with increasing disease duration. These findings support the suggestion that hippocampal atrophy in AD is as a result of neuron loss and confirm the usefulness of volumetry as an indirect measure of neurodegeneration in this disease. PMID- 15135881 TI - Diseases of modern living: neurological changes associated with mobile phones and radiofrequency radiation in humans. AB - Health effects of radiofrequency radiations (RFR) including mobile phone technology and the adequacy of their safety standards remain uncertain. Case reports of peripheral neurological effects of RFR describe mainly disturbances of noxious sensation (dysaesthesia). Cases associated with other RFR sources as well as mobile phone technology are examined seeking insights into neurophysiological mechanisms and safety levels. Cases have arisen after exposure to much of the frequency range (low MHz to GHz). In some instances symptoms are transitory, but may be lasting in others. After very high intensity exposures nerves may be grossly injured. However, after lower intensity exposures which may result in dysaesthesia, ordinary nerve conduction studies demonstrate no abnormality although current perception threshold studies may. Only a small proportion of similarly exposed persons develop symptoms. The role of modulations (e.g. pulses) needs clarification. Some of these observations are not consistent with the prevailing hypothesis that all health effects of RFR arise from thermal mechanisms. It is concluded that RFR from mobile phones can cause peripheral neurophysiological changes in some persons. The effects occur at exposure levels below the present safety levels for RFR. Possible non-thermal mechanisms are discussed and may point to future directions of research. PMID- 15135882 TI - Distribution of cerebral atrophy assessed by magnetic resonance imaging reflects patterns of neuropsychological deficits in Alzheimer's dementia. AB - Neuropsychological deficits were investigated with respect to regional distribution of cerebral atrophy as assessed by volumetric magnetic resonance imaging (MRI) in 50 patients with Alzheimer's dementia (AD; NINCDS-ADRDA criteria) and 20 healthy volunteers. When compared between groups, test performance of all investigated neuropsychological domains including declarative memory, language, praxia, psychomotor speed, as well as attention and concentration was significantly impaired. These deficits were differentially correlated with regional atrophic changes. In particular, volumes of the right amygdala-hippocampus complex correlated with declarative memory performance, whereas volumes of the left temporo-parietal regions correlated with performance in naming and praxia. Furthermore, left frontal lobe atrophy was associated with verbal fluency. Our data confirm the central role that medial temporal atrophy plays for declarative memory deficits in AD and indicate that additional changes in the parietal, temporal and frontal lobes are responsible for further neuropsychological deficits characteristic of this disorder. PMID- 15135883 TI - Long-lasting motor cortex disinhibition after short transient ischemic attacks (TIAs) in humans. AB - Lesion-induced cortical hyperexcitability has been demonstrated in animal models of cerebral ischemia and after human stroke. We used transcranial magnetic stimulation to investigate motor cortex excitability in ten patients who suffered short transient ischemic attacks (TIAs; i.e. duration <60 min) in the week before examination. Intracortical inhibition (ICI) and facilitation (ICF) were assessed using paired-pulse stimulation. Single-pulse stimulation was applied to investigate cortical silent period and transcallosal inhibition. The side affected by the TIA was compared to the normal side of each patient. We found ICI significantly reduced, and a trend towards enhanced ICF on the affected side. All other parameters remained normal. Motor cortex disinhibition may occur after short TIAs in spite of morphologically intact brain tissue. Possibly, these functional changes correlate to the protective neurometabolic mechanisms elicited by short episodes of focal ischemia in animal models and in man. PMID- 15135884 TI - Laser evoked potentials for assessing sensory neuropathy in human patients. AB - Sensory neuropathy usually impairs tactile sensations related to large myelinated afferents (Abeta) as well as thermal-pain sense related to small myelinated (Adelta) and unmyelinated (C) afferents. By selectively affecting large or small fibres, some sensory neuropathies may also provoke a dissociated sensory loss. Standard nerve conduction studies and somatosensory evoked potentials assess Abeta-fibre function only. Laser pulses selectively excite free nerve endings in the superficial skin layers and evoke Adelta-related brain potentials (LEPs). From earlier studies and new cases we collected data on 270 patients with sensory neuropathy. LEPs often disclosed subclinical dysfunction of Adelta fibres and proved a sensitive and reliable diagnostic tool for assessing small-fibre function in sensory neuropathy. PMID- 15135885 TI - Leg length inequality in humans: a new neurophysiological approach. AB - In conditions where there is moderate or severe pain, such as in low back pain, sciatica and osteoarthritis affecting the hip, the patient often reports the sensation that the two legs are of unequal length. This sensation cannot be explained as a result of fracture or other trauma that might cause a real change in length of the legs. There is confusion as to whether this 'leg length inequality' causes the pain or vice versa. It is argued here that the sensation occurs when there is pressure on the sensory nerves, especially the dorsal spinal roots. This causes pain but, more relevantly, causes an increased discharge in muscle spindle nerves, this in turn signaling an increased length of the associated muscles. This idea is testable using percutaneous electrodes. PMID- 15135886 TI - Nicotinic receptor mediated stimulation of NO-generation in neurons of rat thoracic dorsal root ganglia. AB - The contribution of nicotinic acetylcholine receptors (nAChRs) to stimulation of NO-production was investigated in isolated rat dorsal root ganglion (DRG) neurons utilizing an NO-sensitive fluorescent indicator 4,5-diaminofluorescein-diacetate (DAF-2DA) and appropriate channel blockers. RT-PCR and immunohistochemical analysis of NOS isoforms in cultured neurons revealed the expression of eNOS in the vast majority of neurons, nNOS in about 5-10%, and iNOS only exceptionally. Application of nicotine resulted in an abrupt increase in DAF-2T fluorescence in 65% of neuronal cell bodies that was fully sensitive to the general nAChR antagonist mecamylamine. Methyllycaconitine reduced the number of nicotine sensitive neurons and the extent of NO-generation. Thus, alpha7- and/or alpha9/10 nAChRs are required for nicotine-induced NO-production in a subpopulation of DRG neurons, and appear to be partially involved in the remaining, larger subpopulation. PMID- 15135887 TI - Activation of the c-Jun transcription factor following neurodegeneration in vivo. AB - Neurodegenerative diseases often result in neuronal cell death, but the molecular mechanisms responsible are not fully understood. The expression and activation by phosphorylation of the c-Jun transcription factor plays an important role for the fate of affected neurons in response to injury. c-Jun is phosphorylated at serines 63 and 73 by the c-Jun N-terminal kinases and c-Jun N-terminal phosphorylation augments the transcriptional activity of c-Jun. Two approaches in neurodegeneration were investigated: The transection of the medial forebrain bundle to study neuronal cell body response in the derived neuronal populations of the substantia nigra pars compacta (SNC). This model of central axotomy leads as a long-term reaction to degeneration of the affected SNC neurons. A central component of the axotomy-induced alterations leading to neuronal degeneration is the rapid induction, lasting expression and activation of the c-Jun transcription factor. The focal cerebral ischemia, induced by occlusion of the arteria cerebri media and the subsequent reperfusion, serves as a suitable in vivo model for stroke. Also, ischemia leads to upregulation and activation of c-Jun and its target genes. Here the key role of c-Jun for the fate of neurons following degeneration is discussed with data received from experiments performed in Manfred Zimmermann's department investigating the effects of c-Jun on its target genes and on factors influencing c-Jun expression and activation. PMID- 15135888 TI - Interleukin 1alpha and interleukin 6 protect human neuronal SH-SY5Y cells from oxidative damage. AB - Interleukin (IL)-1alpha and IL-6 are powerful inflammatory cytokines produced in brain primarily by microglia and astrocytes. Here we demonstrate, using an in vitro assay system, that they can have a direct neuroprotective action against oxidative attack. Exposure of retinoic acid-differentiated human SH-SY5Y neuroblastoma cells to 270 microM hydrogen peroxide caused activation of caspase 3 and significant neuronal death. Treatment with IL-1alpha or IL-6 caused a dose dependent increase in survival as measured by the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide assay. An antibody against single-strand DNA demonstrated many apoptotic neuroblastoma cells following exposure to hydrogen peroxide, with a decrease following cytokine treatment. These data indicate that IL-1alpha and IL-6 can, under appropriate circumstances, protect neurons from oxidative damage in addition to their well-known action of stimulating inflammation. PMID- 15135889 TI - Molecular determinants of two neurotoxins that regulate sodium current inactivation in rat hippocampal neurons. AB - We studied functional and structural differences between the two neurotoxins, wasp toxin pompilidotoxin (PMTX) and sea anemone toxin (ATXII). Although PMTX and ATXII inhibited inactivation of sodium currents both toxins had distinct actions on the lobster axon and on the rat hippocampal cells. To determine structural basis of the difference we compared arrangement of polar and non-polar amino acids of the two toxins and found that similar sequence of PMTX exist in a discrete position of three-dimensional structure of ATXII. The sequence may be responsible for the binding site in the neuronal Na(+) channel molecule because PMTX is insensitive to cardiac Na(+) channel. Differential actions of ATXII from PMTX may come from other regions than the overlapped sequence. PMTX has diverse actions in the central neurons and is useful to classify Na(+) channel subtypes. PMID- 15135890 TI - Cdk5: mediator of neuronal death and survival. AB - Cdk5 (cyclin-dependent kinase 5) is a serine/threonine kinase implicated to play pivotal roles in neuronal development. Recently, its potential involvement as a regulator of neuronal death and survival has attracted considerable interests. Importantly, increasing evidence has linked Cdk5 to the etiopathology of neurodegenerative diseases such as Alzheimer's disease and amyotrophic lateral sclerosis. Here we summarize the recent findings on Cdk5 not only as an important participant in neuronal death, but also a key player in neuronal survival. Elucidating the mechanisms of regulation of Cdk5 and its downstream signaling might prove to be crucial in the therapeutic treatment of neurodegenerative diseases. PMID- 15135891 TI - GDNF promotes neuronal differentiation and dopaminergic development of mouse mesencephalic neurospheres. AB - In the present study we report establishment of a neurosphere culture system derived from mouse ventral mesencephalon at embryonic day 12 and investigate effects of glial cell line-derived neurotrophic factor (GDNF) in the differentiation potential of the neurospheres. The generated neurospheres exhibit stem cell characteristics, i.e. self-renewal capacity and multipotency. Addition of exogenous GDNF resulted in neural differentiation indicated by reduced number of nestin positive cells. GDNF treatment resulted in increased numbers of beta III-tubulin immunoreactive cells whereas glial fibrillary acidic protein immunoreactivity was not effected. Most importantly, cell numbers expressing early dopaminergic markers, Nurr1 and Ptx3, were significantly higher in GDNF treated spheres. We conclude that GDNF promotes differentiation of mouse mesencephalic stem cells towards neuronal lineage and most notably dopaminergic development. PMID- 15135892 TI - Comparative effects of huperzine A, donepezil and rivastigmine on cortical acetylcholine level and acetylcholinesterase activity in rats. AB - The cholinesterase inhibitors huperzine A, donepezil and rivastigmine were compared for their effects on extracellular acetylcholine concentration and acetylcholinesterase activity in the rat cortex. After i.p. injection, huperzine A (0.25-0.75 micromol/kg), donepezil (2-6 micromol/kg) and rivastigmine (0.75-1.5 micromol/kg) dose-dependently elevated the concentration of acetylcholine. The duration of huperzine A was longest. The time courses of cortical acetylcholinesterase inhibition with middle doses of these agents mirrored the increases of acetylcholine at the same doses. However, acetylcholinesterase inhibition was disproportionately greater after middle dose of rivastigmine than doses of huperzine A and donepezil that increased acetylcholine to a similar extent. Muscle fasciculation appeared only after donepezil with a dose-dependent incidence and intensity. In molar terms, huperzine A was 8- and 2-fold more potent than donepezil and rivastigmine, respectively, in increasing cortical acetylcholine levels, with a longer-lasting effect. PMID- 15135893 TI - Post-synaptic N-methyl-d-aspartate signalling in hippocampal neurons of rat: spillover increases the impact of each spike in a short burst discharge. AB - High-frequency burst discharges in hippocampus typically consist of less than ten spikes fired at frequencies too high to be followed by a post-synaptic neuron. How significant are these numbers for synaptic signalling? We have measured the N methyl-d-aspartate (NMDA) component of the excitatory post-synaptic current (EPSC(NMDA)) in hippocampal CA1 neurons of rat after burst discharge of variable duration. The synaptic facilitation is accompanied by a slow-down of the EPSC(NMDA) which develops on a spike-to-spike basis. Consequently the charge transferred by the after-burst EPSC(NMDA) is increased with each spike. The phenomenon is most probably due to the spillover-mediated recruitment of extrasynaptic NMDA receptors. In terms of post-synaptic signalling it dramatically increases the impact of each spike in a short burst discharge. PMID- 15135894 TI - Neurodegenerative and physiological actions of c-Jun N-terminal kinases in the mammalian brain. AB - The research in the field of AP-1 transcription factor expression, such as Jun or Fos proteins, in the brain was a milestone in neurosciences. The last years have provided growing insights into the upstream signal transduction which controls the expression and activation of these transcriptional regulators. In particular, the c-Jun N-terminal kinases (JNKs) were considered to confer degeneration by activation of c-Jun. Recent findings, however, demonstrate an essential physiological role of JNKs in the nervous system. Here we review the specific control and dual functions of JNK isoforms which are relevant for the development of the intact brain on the one hand, and which can confer dramatic neurodegenerative effects and microglial activation on the other hand. PMID- 15135895 TI - Opposite effects of gamma(1)- and gamma(2)-melanocyte stimulating hormone on regulation of the dopaminergic mesolimbic system in rats. AB - By use of the brain microdialysis technique we show that administration of gamma(1)-melanocyte stimulating hormone (gamma(1)-MSH) into the ventral tegmental area of anaesthetized rats causes an increase in the release of extracellular dopamine and its metabolite 3,4-dihydroxyphenylacetic acid in the nucleus accumbens, while gamma(2)-MSH causes the opposite effect. Moreover, gamma(2)-MSH pre-treatment considerably reduced the gamma(1)-MSH-induced effects. Our findings suggest an opposing action of two gamma-MSH-activated pathways on the mesolimbic dopaminergic system, which could be important in the maintenance of a balanced psychoactivation state. PMID- 15135896 TI - Interaction of gamma-aminobutyric acid receptor type B receptors and calcium channels in nociceptive transmission studied in the mouse hemisected spinal cord in vitro: withdrawal symptoms related to baclofen treatment. AB - An in vitro mouse hemisected spinal cord was used to characterize the gamma aminobutyric acid receptor type B (GABA(B)) modulation of the ventral root potential (VRP) in response to electrical stimulation of the dorsal root (DR). Low-intensity (LI) and high-intensity (HI) stimulation induced VRPs with progressively higher amplitude and duration. Repetitive HI-stimulation of the DR (1-10 Hz) produced windup. The selective GABA(B) receptor agonist, CGP35024, inhibited the VRPs in a dose-dependent manner. The inhibitory action of CGP35024 was blocked by CGP52432, a potent GABA(B) receptor antagonist. Following washout of the GABA(B) receptor agonist, VRPs and windup were significantly enhanced. The rebound increase of the VRP following removal of CGP35024 was also blocked by the GABA(B) receptor antagonist, CGP52432. This phenomenon is not linked to receptor desensitization, but rather due to GABA(B) receptor-induced hyperactivity of N-, P/Q-type Ca(2+) channels, as omega-CgTx GVIA and MVIIC abolished/prevented the increase. The 'rebound' enhancement of the spinal transmission after exposure to GABA(B) agonists sheds light on the possible mechanism of the severe withdrawal effects after abrupt termination of baclofen treatment in patients suffering from multiple sclerosis. PMID- 15135897 TI - Nervous system injury: focus on the inflammatory cytokine 'granulocyte-macrophage colony stimulating factor'. AB - Any lesion in the nervous system, be it infectious, immunopathological, ischemic or traumatic, is followed by an inflammatory process that induces rapid activation of glial cells and additional recruitment of granulocytes, T-cells and monocytes/macrophages from the blood stream. Neuroinflammation is a double-sided sword. It can cause neuronal damage and participate in neuropathic pain, but it also has neuroprotective and neurotrophic effects at some stages. Cytokines are the main molecular actors of this 'network of inflammation'. Among them, granulocyte-macrophage colony-stimulating factor (GM-CSF) is a pro-inflammatory hematopoietic cytokine widely used in haematological disorders to stimulate proliferation and differentiation of neutrophilic, eosinophilic and monocytic lineages. GM-CSF and its receptor are expressed in the brain and the cytokine can cross the blood-brain barrier. It is thus likely to affect various nervous system functions. This review will focus on the role of GM-CSF in nervous system disorders and their experimental models with particular emphasis on its possible beneficial effect on axonal regeneration after PNS and CNS injury. PMID- 15135898 TI - Spinal c-fos expression associated with spontaneous biting in a mouse model of dry skin pruritus. AB - As a model of dry skin pruritus in mice, one hind paw was treated twice daily with a mixture of acetone/diethylether/water (AEW); controls received water only. A protective collar prevented the animals from accessing the treatment area. At 16 days, the collar was removed and AEW-treated mice exhibited marked biting of the treated paw; the number and cumulative duration of bites was significantly greater than in controls. After 3 additional treatment days (collars intact), animals were perfused for c-fos immunohistochemistry. There was significantly more fos-like immunoreactivity in the ipsilateral lumbar spinal cord of AEW treated animals, with the majority in superficial laminae. It is proposed that biting of the dry skin reflects pruritus, and that neurons predominantly in superficial laminae of the dorsal horn may signal itch sensation. PMID- 15135899 TI - Three bands of oscillatory activity in the lateral geniculate nucleus of the cat visual system. AB - Neuronal responses in the cat lateral geniculate nucleus were analyzed with respect to oscillatory components in 5930 peri-stimulus time histograms recorded in 69 neurons. Oscillatory components were observed in three frequency ranges clearly separated from each other, i.e. in a lower range between 25 and 33 Hz (mean 27.8 Hz), in a middle range between 45 and 60 Hz (mean 52.5 Hz), and in a higher range between 75 and 100 Hz (mean 84.4 Hz); temporal variance within the neuronal populations showing oscillatory characteristics increased with higher frequencies. Although the frequency ranges of the oscillatory responses were clearly separated from each other, a functional dependence between these three populations of geniculate neurons appears to be possible; the numerical relationship of approximately 1:2:3 points to the possibility of a temporal coupling between distinct neuronal populations at an early stage of visual processing. PMID- 15135900 TI - Longterm stability and developmental changes in spontaneous network burst firing patterns in dissociated rat cerebral cortex cell cultures on multielectrode arrays. AB - Spontaneous action potentials were recorded longitudinally for 4-7 weeks from dissociated rat occipital cortex cells cultured on planar multi-electrode plates, during their development from isolated neurons into synaptically connected neuronal networks. Activity typically consisted of generalized bursts lasting up to several seconds, separated by variable epochs of sporadic firing at some of the active sites. These network bursts displayed discharge patterns with age dependent firing rate profiles, and durations significantly increasing in the 3rd week in vitro and decreasing after about 1 month in vitro, when they evolved into short events with prompt onsets. These findings indicate that after about a month in vitro these cultured neuronal networks have developed a degree of excitability that allows almost instantaneous triggering of generalized discharges. Individual neurons tend to fire in specific and persistent temporal relationships to one another within these network bursts, suggesting that network connectivity maintains a core topology during its development. PMID- 15135901 TI - Activation of the intracerebral cholinergic nerve fibers originating in the basal forebrain increases regional cerebral blood flow in the rat's cortex and hippocampus. AB - In the rat, activation of the intracerebral cholinergic system originating in the basal forebrain and projecting to the cortex and hippocampus releases acetylcholine in the cortex and hippocampus, which results in vasodilation and an increase in regional cerebral blood flow (rCBF) in the cortex and hippocampus. The augmentation of rCBF is independent of both systemic blood pressure and regional metabolism. The intracerebral cholinergic fibers are able to act as autonomic nerve fibers for the regulation of cortical and hippocampal blood flow. PMID- 15135902 TI - Preserved reticular neuronal activity during selective delivery of supra-clinical isoflurane concentrations to brain in goats and its association with spontaneous movement. AB - We have previously observed spontaneous movement when supra-clinical concentrations of isoflurane were selectively delivered to the in situ goat brain. We presently examined whether neurons in the midbrain reticular formation (MRF) remained active during such delivery. Isoflurane (5.1+/-1.6%) was selectively delivered to the goat cranial circulation while the torso isoflurane was decreased to 0.2-0.3%. At the high cranial/low torso isoflurane combination, spontaneous movement occurred that usually consisted of running motions, stiffening and twitching. Seventeen MRF neurons recorded in six goats exhibited high spontaneous firing (33+/-20 Hz) despite the presence of an isoelectric electroencephalogram. We conclude that some MRF neurons are resistant to the depressant effect of isoflurane, and may contribute to the observed spontaneous movement. PMID- 15135903 TI - Multiple representation of sensory system input in the cerebral cortex:the new phrenology? PMID- 15135904 TI - Effects of spinal and peripheral nerve lesions on the intersegmental synchronization of the spontaneous activity of dorsal horn neurons in the cat lumbosacral spinal cord. AB - In the anesthetized and paralyzed cat, spontaneous negative cord dorsum potentials (nCDPs) appeared synchronously in the L3 to S1 segments, both ipsi- and contralaterally. The acute section of both the intact sural and the superficial peroneal nerve increased the variability of the spontaneous nCDPs without affecting their intersegmental coupling. On the other hand, the synchronization between the spontaneous nCDPs recorded in segments L5-L6 was strongly reduced following an interposed lesion of the left (ipsilateral) dorsolateral spinal quadrant and it was almost completely abolished by an additional lesion of the contralateral dorsolateral quadrant at the same level. Our observations support the existence of a system of spontaneously active dorsal horn neurons that is bilaterally distributed along the lumbosacral segments and affects, in a synchronized and organized manner, impulse transmission along many reflex pathways, including those mediating presynaptic inhibition. PMID- 15135905 TI - Oscillatory activity in forelimb muscles of behaving monkeys evoked by microstimulation in the cerebellar nuclei. AB - Coherent 20-35 Hz (beta) oscillations are a prominent feature of activity in primary motor cortex and muscles of monkeys and humans performing voluntary movements. We found that coherent beta oscillations are also present in the cerebellar nuclei (CN). Two monkeys were operantly conditioned to perform a wrist flexion/extension step-tracking task while we recorded neuronal activity or microstimulated in CN and recorded EMG activity from forelimb muscles. Coherent beta oscillations were found between discharges of some CN neurons and tonically active shoulder, elbow and wrist/finger flexion and extension muscles. Similarly, localized microstimulation pulses in CN evoked transient beta oscillations in widespread forelimb muscles. We conclude that coherent motor system beta oscillations are present in CN and that CN may be an important nodal point for the generation and/or propagation of beta oscillations throughout the motor system. PMID- 15135906 TI - Brain development during fetal life: influences of the intra-uterine environment. AB - The intrauterine environment can significantly affect fetal brain development. Here we review our recent findings using animal models that mimic adverse intrauterine conditions which could exist during human pregnancy. We have focused on effects of both acute and chronic hypoxic and inflammatory insults. Relatively brief periods of hypoxemic compromise can have significant effects on the fetal brain causing neuronal loss and cerebral white matter damage. Subtle brain injury can occur, for example to a particular class of neuron, and this can have a significant effect on the function of a specific system. Chronic mild placental insufficiency can result in long term deficits in neuronal connectivity affecting function postnatally as demonstrated in the auditory and visual systems. Repeated acute exposure to an inflammatory agent results in diffuse subcortical white matter damage and in some cases periventricular necrosis. We have demonstrated that the timing and severity of these prenatal insults are determinants of the outcomes, in terms of the severity of the damage and the regions of the brain affected. PMID- 15135907 TI - Efficacy of sonothrombolysis in a rat model of embolic ischemic stroke. AB - The key goal in the treatment of acute ischemic stroke is fast vessel recanalization. Thrombolysis with recombinant tissue plasminogen activator (rt PA) is efficient in humans but mean time for recanalization is within hours. Ultrasound bio-effects has been shown to facilitate rt-PA mediated thrombolysis in peripheral arteries. We used an embolic stroke model in the rat. In all rats we induced an ischemic stroke by a selective occlusion of the middle cerebral artery with whole blood clots. From an entire collective of 54 rats 47 completed the protocol (n = 7 died early). Four different groups (no treatment n = 6; full dose rt-PA treatment only [10 mg/kg per body weight] n = 14, half dose rt-PA treatment plus ultrasound n = 10, and full dose rt-PA treatment plus ultrasound n = 17) were investigated. We found a significant reduction of absolute as well as relative infarct volume in the full dose rt-PA plus ultrasound group (81+/-72 mm(3); P< 0.05) in comparison to untreated rats (253+/-159 mm(3); P < 0.05) as well as in comparison to rats treated with full dose rt-PA only (167+/-91 mm(3); P < 0.05). There were five intracranial bleedings giving a bleeding rate of 9.3%. In summary: ultrasound treatment in addition to rt-PA is more effective than single rt-PA treatment in reducing infarct volume and safe with regard to bleeding. PMID- 15135908 TI - S 18986, a positive modulator of AMPA receptors with cognition-enhancing properties, increases ACh release in the hippocampus of young and aged rat. AB - The effect of S 18986, positive AMPA receptor modulator, on acetylcholine (ACh), gamma-aminobutyric acid (GABA) and glutamate (Glu) release from the hippocampus of freely moving young and aged rats was investigated by microdialysis coupled to HPLC. The cognition-enhancing properties were evaluated by a passive avoidance test. In 3 month-old rats, S 18986 (10 mg/kg i.p.) increased by 70% ACh release, which returned to basal level within 2 h, while 3 mg/kg had no effect. In 22 month-old rats, both 3 and 10 mg/kg i.p. induced a long lasting increase in ACh release, as large as that induced by 10 mg/kg in young rats. S 18986 did not modify GABA and glutamate release. No effect on general behavior was observed, but S 18986 at both doses prevented the disrupting effect of scopolamine (1 mg/kg i.p.) on passive avoidance acquisition. PMID- 15135909 TI - Neuropeptide research discloses part of the secrets of Alzheimer's disease neuropathogenesis: state of the art 2004. AB - Molecular misreading, a process discovered in the late 1990s, entails the formation of aberrant transcripts due to the inaccurate conversion of genomic information, and results in an accumulation of aberrant proteins. The aberrant transcripts are formed as a result of a dinucleotide deletion (e.g. DeltaGA, DeltaGU) during or after transcription. Either the RNA polymerase starts to make mistakes (e.g. stuttering) in simple sequence repeats, such as GAGAG, or erroneous editing of transcripts occurs. If these aberrant transcripts are not detected and degraded efficiently, they can be translated from the deletion onwards into the +1 reading frame. The resulting proteins are therefore called +1 proteins. If functional domains are located downstream of the frameshift site, the result will be a protein with a potential loss or gain of function. It has been hypothesized that quality control mechanisms for both transcripts and proteins work less efficiently during aging, which is why +1 proteins may become manifest and contribute to age-related diseases in neuronal and non-neuronal cells. PMID- 15135910 TI - Intranasal administration of NAP, a neuroprotective peptide, decreases anxiety like behavior in aging mice in the elevated plus maze. AB - NAP, an 8-amino-acid peptide (NAPVSIPQ=Asn-Ala-Pro-Val-Ser-Ile-Pro-Gln), provides neuroprotection at very low doses in a variety of animal models. Previously, acute NAP administration by the intranasal route resulted in improved performance in the Morris water maze of normal and cognitively impaired rats. In these animals, it was observed, but not quantified, that NAP exhibited an anxiolytic effect. Therefore, we have tested here the effects of chronic NAP treatment on anxiety-like behavior in mice in the elevated plus maze. Results showed that 5 months of daily (intranasal) treatment with NAP reduced anxiety, measured as the percentage of time spent in the open arms of the maze (P < 0.01). This effect was maintained after a longer (8 months) exposure to NAP. In addition, after 8 months of NAP treatment, the percentage of open arm entries out of total arms entries was significantly higher in the treated mice ( P < 0.01). Motor function indices indicated no significant differences between the groups. Furthermore, prolonged treatment with NAP (7 months) showed some beneficial effects on Morris water maze performance in the aging mice. It is concluded that NAP offers a unique combination of anxiolytic/cognitive enhancing properties observed after prolonged chronic intranasal treatment. PMID- 15135911 TI - SRA880, in vitro characterization of the first non-peptide somatostatin sst(1) receptor antagonist. AB - This report describes the in vitro features of the first somatostatin sst(1) receptor selective non-peptide antagonist, SRA880 ([3R,4aR,10aR] 1,2,3,4,4a,5,10,10a-Octahydro-6-methoxy-1-methyl-benz[g] quinoline-3-carboxylic acid-4-(4-nitro-phenyl)-piperazine-amide, hydrogen malonate). SRA was evaluated in a number of in vitro systems of various species, both at native and recombinant receptors, using radioligand binding and second messenger/transduction studies. SRA880 has high affinity for native rat, mouse, monkey and human cerebral cortex somatostatin sst(1) receptors (pK(d) = 7.8-8.6) and for human recombinant sst(1) receptors (pK(d) = 8.0-8.1). SRA880 displayed significantly lower affinity for the other human recombinant somatostatin receptors ( pK(d) < or = 6.0) or a wide range of neurotransmitter receptors, except for the human dopamine D4 receptors. SRA880 was characterized in various transduction assays: somatotropin release inhibiting factor (SRIF) induced inhibition of forskolin-stimulated cAMP accumulation, SRIF stimulated-GTPgammaS binding, and SRIF stimulated luciferase gene expression; in all tests, SRA880 was devoid of intrinsic activity and acted as an apparently surmountable antagonist with pK(B) values of 7.5-7.7. Combined with the data from binding studies, these results suggest that SRA880 acts as a competitive antagonist. Thus, SRA880 is the first non-peptide somatostatin sst(1) receptor antagonist to be reported; SRA880 will be a useful tool for the characterization of somatostatin sst(1) receptor mediated effects both in vitro and in vivo. PMID- 15135912 TI - Secondary tactile hypoesthesia: a novel type of pain-induced somatosensory plasticity in human subjects. AB - Quantitative sensory testing revealed that pain induced by intracutaneous capsaicin injection elicited secondary hyperalgesia coexisting with secondary tactile hypoesthesia. Mapping the areas of altered mechanical sensations adjacent to the capsaicin injection disclosed that the area of secondary hyperalgesia was always nested in a larger area of secondary hypoesthesia easily detected as numbness by most subjects. Psychometric functions revealed a twofold rightward shift of tactile detection (hypoesthesia), which coexisted with a more than fourfold leftward shift of pricking pain detection (hyperalgesia) in the same skin area. As a mechanism we propose a functional switch at the spinal level based on C-fibre-induced primary afferent depolarisation resulting in presynaptic inhibition of low threshold mechanoreceptor input and an ensuing loss of tactile sensitivity. PMID- 15135913 TI - Spatial integration of cold pressor pain sensation in humans. AB - Spatial integration of cold pressor pain (CPP) in the hand was studied in healthy human subjects by measuring the latency to the ice water-induced first pain sensation with and without conditioning CPP. CPP alone showed a marked spatial summation effect. When conditioning and test CPP were applied at the same time, conditioning CPP suppressed test CPP both in an adjacent and a distant site. When test CPP was applied after the conditioning CPP (i.e. pain induced by conditioning CPP was considerably stronger than that evoked by test CPP) conditioning CPP suppressed the test CPP only in a distant site but enhanced it in an adjacent site. A decrease in the test stimulus area increased the suppressive effect by conditioning CPP. Thus, CPP shows spatial summation or inhibition depending on experimental parameters. PMID- 15135914 TI - Methodologic factors which contribute to variations in experimental pain threshold reported for older people. AB - Using the same study groups and psychophysical methods, we have tested the hypothesis that variations in pain threshold with advancing age are best explained by variations in stimulus duration. Fifteen young adults and 15 older people without clinical evidence of neurologic disease or psychologic dysfunction had pain thresholds determined with heat and electrical stimuli using the method of limits; for electrical stimulation a double random staircase design was used. The stimulus duration was 1-100 s for heat and 50-5000 ms for electrical stimulation. It was found that older people have an increased threshold for thermal and electrically induced pain if the stimulus duration is kept short. This result explains much of the variability in age associated pain threshold in the literature. PMID- 15135916 TI - Classification of psychosocial risk factors (yellow flags) for the development of chronic low back and leg pain using artificial neural network. AB - Due to international guidelines of treatment of acute low back pain (LBP), psychosocial risk factors like depressive mood and maladaptive pain-related coping strategies ('yellow flags') have to be assessed in the early phase of acute pain. Within this longitudinal study in patients with LBP and leg pain, we used an artificial neural network (ANN) to classify the pain intensity 6 months after the onset of treatment. Psychosocial risk factors were used as input neurons. The training of the three-layer ANN using the back-propagation algorithm yields to an accuracy of 83.1%. Further on, the complexity of this structure indicated the necessity of an early screening procedure that allows a differentiation between several high risk groups and a low risk profile in order to predict the long-term development of pain intensity. PMID- 15135915 TI - Peripheral and electrocortical responses to painful and non-painful stimulation in chronic pain patients, tension headache patients and healthy controls. AB - Sixteen chronic back pain (CBP) patients, 16 tension headache (THA) patients and 16 healthy controls (HC) were exposed to four series of ten electric stimuli at perception threshold, pain threshold and 10% below pain tolerance. The EEG was recorded from three sites, in addition, the EMG from the m. frontalis and m. erector spinae, heart rate and skin conductance were assessed. The CBP patients showed significantly lower pain threshold and pain tolerance values than the HC and the THA patients whereas the THA patients displayed a higher pain tolerance. Habituation was reduced in the CBP group. N150, P260, P300 and N500 were not significantly different between the groups nor were there significant group differences in the peripheral measures. However, since the stimulation intensity was significantly lower in the CBP patients, these data are indicative of both enhanced central and peripheral reactivity. The observed lack of habituation may contribute to the persistence of chronic pain. PMID- 15135917 TI - Direct evidence for activation and desensitization of the capsaicin receptor by N oleoyldopamine on TRPV1-transfected cell, line in gene deleted mice and in the rat. AB - Effects of the endogenous lipid N-oleoyldopamine (OLDA) were analyzed on the rTRPV1-expressing HT1080 human fibrosarcoma cell line (HT5-1), on cultured rat trigeminal neurons, on the noxious heat threshold of rats and on nocifensive behavior of TRPV1 knockout mice. The EC(50) of capsaicin and OLDA on (45)Ca accumulation of rTRPV1-expressing HT5-1 cells was 36 nM and 1.8 microM, respectively. The efficacy of OLDA was 60% as compared to the maximum response of capsaicin. OLDA (330 nM to 3.3 microM) caused a transient increase in fluorescence of fura-2 loaded cultured small trigeminal neurons of the rat and rTRPV1-transfected HT5-1 cells measured with a ratiometric technique. Repeated application of OLDA and capsaicin caused similar desensitization in the Ca(2+) transients both in cultured neurons and rTRPV1-transfected HT5-1 cells. In the rat intraplantar injection of OLDA (5 nmol) decreased the noxious heat threshold by 6-9 degrees C and this response was strongly inhibited by the TRPV1 antagonist iodoresiniferatoxin (0.05 nmol intraplantarly (i.pl.)). In wild-type mice OLDA (50 nmol i.pl.) evoked paw lifting/licking which was significantly less sustained in TRPV1 knockout mice. It is concluded that on TRPV1 capsaicin receptors OLDA is 50 times less potent than capsaicin and it might serve as an endogenous ligand for TRPV1 in the rat, but more likely in humans. PMID- 15135918 TI - Histamine-induced Ca(2+) influx via the PLA(2)/lipoxygenase/TRPV1 pathway in rat sensory neurons. AB - Histamine is known to excite a subset of C-fibers and cause itch sensation. Despite its well-defined excitatory action on sensory neurons, intracellular signaling mechanisms are not understood. Previously, we demonstrated that bradykinin excited sensory neurons by activating TRPV1 via the phospholipase A(2) (PLA(2)) and lipoxygenase (LO) pathway. We, thus, hypothesized that histamine excited sensory neurons via the PLA(2)/LO/TRPV1 pathway. Application of histamine elicited a rapid increase in intracellular Ca(2+) ([Ca(2+)](i)) that desensitized slowly in cultured dorsal root ganglion neurons. Histamine-induced [Ca(2+)](i) was dependent on extracellular Ca(2+) and inhibited by capsazepine and by SC0030, competitive antagonists of TRPV1. Quinacrine and nordihydroguaiaretic acid, a PLA(2) and an LO inhibitor, respectively, blocked the histamine-induced Ca(2+) influx in sensory neurons, while indomethacin (a cyclooxygenase inhibitor) did not. We thus conclude that histamine activates TRPV1 after stimulating the PLA(2)/LO pathway, leading to the excitation of sensory neurons. These results further provide an idea for potential use of TRPV1 antagonists as anti-itch drugs. PMID- 15135919 TI - Responsiveness of C-fiber nociceptors to punctate force-controlled stimuli in isolated rat skin: lack of modulation by inflammatory mediators and flurbiprofen. AB - Although cutaneous C-fiber nociceptors show dramatic inflammatory sensitization to heat, they do not appear to get sensitized to the mechanical stimulation by von Frey hairs. We employed force-controlled punctate electromechanical stimulation to receptive fields of 61 characterized C-fibers in the isolated rat skin-saphenous nerve preparation. In general: low-in contrast to higher-threshold units showed greater dynamic sensitivity and response magnitude, an earlier onset and a stronger degree of adaptation, the latter due to the linear rise of the force stimulus. On this methodological basis three groups of units were subject to a mix of inflammatory mediators, to flurbiprofen or to control solution. Subsequent mechanostimulation revealed a good reproducibility of the control response and no significant changes in the treatment groups. In conclusion, even refined mechanostimulation was unable to demonstrate sensitization of the predominant nociceptor classes in the rat skin. PMID- 15135920 TI - The many functions of nerve growth factor: multiple actions on nociceptors. AB - Nerve growth factor (NGF) initially interested neurobiologists because of its effects in the developing nervous system. It is now clear that NGF functions throughout the life of the animal with a wide repertoire of actions. In the sensory nervous system it primarily influences the structure and function of nociceptors. Here, we provide a brief review of these actions and raise the overriding biological questions of why these multiple actions occur and how they are carried out. PMID- 15135921 TI - The proportion of TRPV1 protein-positive lumbar DRG neurones does not increase in the course of acute and chronic antigen-induced arthritis in the knee joint of the rat. AB - The TRPV1 receptor, previously called VR1 receptor, is a non-selective cation channel gated by capsaicin, noxious heat, protons and anandamide. The TRPV1 receptor is essential for the development of thermal hyperalgesia. The present study investigated whether the proportion of neurones with TRPV1 receptor increases in lumbar DRG neurones in the course of an antigen-induced arthritis (AIA) of one knee joint in the rat. In control rats 38.1+/-2.3% of the neurones from sections of the L1-L5 ganglia showed TRPV1-like immunoreactivity. Neither in the acute (3 days) nor chronic phase (21 days) of AIA in the knee joint the proportion of TRPV1-like immunoreactive profiles showed significant changes. Thus AIA in the knee joint is not associated with an up-regulation of the TRPV1 receptor in the lumbar DRG neurones. PMID- 15135922 TI - A role for substance P in arthritis? AB - Substance P is a neuropeptide that is released from sensory nerves and which has a number of pro-inflammatory effects. In this article, we review the evidence for a role of substance P in arthritis, both in experimental animal models and rheumatoid arthritis patients. Substance P expression is altered in the joint and dorsal horn of arthritic animals, exogenous substance P and neurokinin 1 (NK(1)) receptor antagonists modulate responses in the joint, and there is some evidence for a role of substance P in human joint disease. However, the therapeutic potential of NK(1) receptor antagonists in the treatment of rheumatoid arthritis remains controversial. PMID- 15135923 TI - Octreotide, a somatostatin analogue, attenuates movement evoked discharges of fine afferent units from inflamed knee joints of rats. AB - This electrophysiological study examined whether octreotide, a stable analogue of somatostatin (SOM), reduces the mechanosensitivity of fine primary afferents from inflamed knee joints of rats similarly to SOM itself (Pain 73 (1997) 377). Close intra-arterial application of 200 microl of octreotide (10(-6)-10(-3) M) dose dependently diminished the responses to passive non-noxious and noxious rotations of the joint in most of the units tested. The inhibitory effects of octreotide required a higher concentration (10(-3) M) compared to SOM to successfully decrease the number of recorded spikes. Application of cyclo-somatostatin, a SOM antagonist, before the octreotide injection prevented the reduction of the movement evoked discharges. These data indicate that octreotide is able to successfully decrease the responses of mechanosensitive fine afferent units innervating the inflamed knee joint of the rat and may, therefore, be useful in the treatment of articular pain of peripheral origin. PMID- 15135924 TI - Recent findings on how proinflammatory cytokines cause pain: peripheral mechanisms in inflammatory and neuropathic hyperalgesia. AB - Numerous experimental studies provide evidence that proinflammatory cytokines induce or facilitate inflammatory as well as neuropathic pain and hyperalgesia. Direct receptor-mediated actions of cytokines on afferent nerve fibers have been reported as well as cytokine effects involving further mediators. The final outcome of cytokine action greatly depends on whether they act in the central of in the peripheral nervous system. Here we summarize recent findings on the peripheral mechanisms of action of three prototypic proinflammatory cytokines, interleukin-1beta, interleukin-6 and tumor necrosis factor-alpha, with regards to pain and hyperalgesia. PMID- 15135925 TI - Peripheral nerve injury evokes disabilities and sensory dysfunction in a subpopulation of rats: a closer model to human chronic neuropathic pain? AB - Chronic pain conditions for which treatment is sought are characterized usually by complex behavioural disturbances as well as pain. We review here evidence that although chronic constriction injury (CCI) of the sciatic nerve evokes allodynia and hyperalgesia in all rats, persistent social behavioural and sleep disruption occurs only in a subpopulation of animals. The finding that the 'degree of pain', as defined by allodynia and hyperalgesia, is the same in all animals suggests that the complex behavioural disabilities are independent of the level of sensory dysfunction. An absence of correlation between disability and sensory dysfunction is characteristic also of human neuropathic pain. These findings indicate that: (i). in a subpopulation of rats sciatic injury evokes disabilities characteristic of human neuropathic pain conditions; and (ii). testing for sensory dysfunction alone cannot detect this subpopulation. PMID- 15135926 TI - Opioids and opioid receptors in the enteric nervous system: from a problem in opioid analgesia to a possible new prokinetic therapy in humans. AB - The gut is a neurological organ, which implies that many neuroactive drugs such as opioid analgesics can seriously disturb gastrointestinal function, because many of the transmitters and transmitter receptors present in the brain are also found in the enteric nervous system. One of the most common manifestations of opioid-induced bowel dysfunction is constipation which results from blockade of peristalsis and intestinal fluid secretion. The discovery of opioid receptor antagonists with a peripherally restricted site of action, such as N methylnaltrexone and alvimopan, makes it possible to normalize bowel function in opiate-treated patients without compromising central opioid analgesia. There is emerging evidence that opioid receptor antagonists may also have prokinetic actions, reversing pathological states of gastrointestinal hypomotility that are due to overactivity of the enteric opioid system. PMID- 15135927 TI - Effects of a chronic myositis on structural and functional features of spinal astrocytes in the rat. AB - The study aimed at the question if astrocytes react with morphological or functional changes when a skeletal muscle is pathologically altered. In rats, a myositis was induced in the gastrocnemius-soleus muscle. After 12 days, the immunoreactivity (IR) for glial fibrillary acidic protein (GFAP), morphometric parameters, and fibroblast growth factor-2 (FGF-2) expression of astrocytes were quantitatively evaluated in the dorsal horn of the spinal segment L4. Following inflammation, the area density of GFAP-IR as well as the proportion of astrocytes expressing FGF-2 increased significantly while the degree of astrocyte arborisation decreased as shown by a shape factor. The density of cell nuclei was unchanged suggesting that no myositis-induced cell divisions occurred. The data indicate that spinal astrocytes may influence pain processes particularly by increased FGF-2 synthesis. PMID- 15135928 TI - Presynaptic inhibition and spinal pain processing in mice: a possible role of the NKCC1 cation-chloride co-transporter in hyperalgesia. AB - We have examined the role of the NKCC1 sodium-potassium-chloride-cotransporter in the generation of touch-evoked pain. The pain behavior of NKCC1 knockout mice (KO) was studied and compared to that of heterozygous (HE) and wild-type (WT) littermates. NKCC1 KO mice showed an increase in tail flick latencies and a reduction of the duration of pain behavior induced by intradermal capsaicin compared to HE and WT mice. All three groups of animals expressed a normal level of plasma extravasation following capsaicin applications. NKCC1 KO mice showed a reduction in stroking hyperalgesia (touch-evoked pain) compared to WT and HE mice but no differences were detected between the three groups in the expression of punctate hyperalgesia. As the NKCC1 co-transporter is responsible for the generation of presynaptic inhibition between afferent terminals in the spinal cord, these results support the notion that presynaptic interactions between low and high threshold afferents can underlie touch-evoked pain. PMID- 15135929 TI - Selective C-fiber deafferentation of the spinal dorsal horn prevents lesion induced transganglionic transport of choleragenoid to the substantia gelatinosa in the rat. AB - The effect of neonatal capsaicin treatment, producing selective elimination of almost all unmyelinated C-fiber sensory axons, was studied on lesion-induced transganglionic labelling of the substantia gelatinosa of the spinal cord by choleragenoid. In both control and capsaicin-pretreated rats, the injection of choleragenoid-horseradish peroxidase conjugate into the intact sciatic nerves resulted in intense labelling only of the deeper layers of the spinal dorsal horn. In the control but not the capsaicin-pretreated rats, the injection of the tracer into sciatic nerves transected 2 weeks previously produced an intense homogeneous labelling of the substantia gelatinosa. It is concluded that the uptake and axonal transport of choleragenoid by capsaicin-sensitive C-fiber afferents may be accounted for by the lesion-induced transganglionic labelling of the substantia gelatinosa, rather than by A-fiber sprouting. PMID- 15135930 TI - Effects of peripheral nerve injury on delta opioid receptor (DOR) immunoreactivity in the rat spinal cord. AB - Morphine and other opioids have direct analgesic actions in the spinal cord and chronic spinal administration of opioid agonists is used clinically in patients suffering from severe, chronic pain. Neuropathic pain resulting from peripheral nerve injury is often less sensitive to opioid therapy than other forms of chronic pain in both humans and animal models. Changes in spinal mu-opioid receptor (MOR) expression have been demonstrated in animal models of neuropathic pain. However, these changes alone fail to account for the attenuation of opioid activity. Reduced expression of delta-opioid receptors (DOR) following peripheral nerve injury has been reported but most of these reports are limited to subjective observation. The magnitude and consistency of these changes is therefore unclear. In addition, previous studies did not evaluate the effects of nerve injury on behavioral measures to confirm induction of aberrant pain symptoms. We therefore performed quantitative image analysis to evaluate the effect of peripheral nerve injury on DOR-immunoreactivity in spinal cord sections from rats previously characterized for sensory responsiveness. We observed statistically significant decreases ipsilateral to nerve injury in all three models tested: sciatic nerve transection, chronic constriction injury of the sciatic nerve and L5/L6 spinal nerve ligation. These results suggest that decreases in the expression of DOR are a common feature of peripheral nerve injury. PMID- 15135931 TI - c-Fos expression in the spinal cord of female rats with artificial ureteric calculosis. AB - Rats with an artificial stone in the left ureter display spontaneous pain behavior (ureteral 'crises') and referred hyperalgesia/contraction in the ipsilateral oblique musculature. To evaluate neuronal activation in both sensitive and motor pathways in this model, c-Fos expression was studied in the spinal cord of calculosis rats vs. sham controls. Fos-labeled cells were never observed in sham controls. In stone rats, they were found in the T10-L2 segments, throughout the dorsal horn, significantly more on the left than the right side (P < 0.002). Fos-labeled cells were also found in lamina IX, containing motoneurons; at the T11-T12 level, these were significantly more on the left than the right side (P < 0.03). Nociceptive input from the ureter thus activates not only sensory but also efferent neurons in the spinal cord, suggesting the triggering of reflex arcs by the visceral focus. PMID- 15135932 TI - Substance P release in the cat spinal cord upon afferent C-fibre stimulation is not attenuated by clonidine at analgesic doses. AB - In anaesthetized cats, antibody microprobes were used to measure the release of immunoreactive substance P (irSP) in the lumbar dorsal horn during electrical stimulation of primary afferent fibres at intensities suprathreshold for unmyelinated fibres. Release of irSP was detected in the region of the superficial dorsal horn. This evoked release was not reduced by clonidine hydrochloride, administered intravenously or by superfusion of the dorsal cord surface. Microprobes inserted during cord superfusion with lignocaine hydrochloride detected less irSP along their entire length, including in the region of evoked release. The results suggest that the analgesic action of clonidine does not involve reduced release of SP from the central terminals of nociceptors in the spinal cord. PMID- 15135933 TI - Activation of presynaptic group I metabotropic glutamate receptors enhances glutamate release in the rat spinal cord substantia gelatinosa. AB - The activation of group I metabotropic glutamate receptors (mGluRs) produces a long-term potentiation of sensory transmission in the substantia gelatinosa (SG) region of the spinal cord (Prog. Brain Res. 129 (2000) 115). The mechanism(s) responsible for the induction of this potentiation is not known. Using rat spinal cord slice preparation and patch-clamp recordings, here we show, that the activation of the group I mGluRs by (S)-3,5-dihydroxyphenylglycine (DHPG, 1 microM), the mGluR1/5 agonist, increased the frequency of both activity-dependent spontaneous EPSCs, and activity-independent miniature EPSCs (mEPSCs). However, DHPG did not affect amplitude of mEPSCs. The effects of DHPG were not seen in the presence of the preferential mGluR1 antagonist CPCCOEt (10 microM). On the other hand, 2-methyl-6-(phenylethynyl)-pyridine (10 microM), a selective mGluR5 antagonist, blocked the DHPG facilitation present during the wash-out of the drug. This novel facilitating effect of the group I mGluR activation on glutamate release is the first report of a direct facilitatory action of both mGluR1 and mGluR5 subtypes on sensory transmission in the spinal cord SG region. These results indicate the potential contribution of synaptic activation of these facilitatory autoreceptors in plasticity of primary afferent neurotransmission. PMID- 15135934 TI - To the descending pain-control system in rats, inflammation-induced primary and secondary hyperalgesia are two different things. AB - The periaqueductal gray matter and the rostral ventromedial medulla (RVM), with its projections to the spinal dorsal horn, constitute the efferent channel of the 'descending pain-control system'. Noxious stimulation of a peripheral tissue causes more pain if this tissue is inflamed (primary hyperalgesia). In such cases, stimulation of neighboring but uninflamed tissues also becomes more painful (secondary hyperalgesia). In animal models of inflammation, the descending pain-control system sends down, simultaneously, inhibitory and facilitatory influences, but inhibition predominates for primary hyperalgesia while facilitation predominates for secondary hyperalgesia. Descending inhibition and facilitation during peripheral inflammation are due not only to previously existing descending modulation, but also to inflammation-induced changes in RVM which involve receptors for NMDA, AMPA, cholecystokinin and neurotensin, as well as synthesis of enkephalins and nitric oxide. PMID- 15135935 TI - Descending serotonergic facilitation mediated through rat spinal 5HT3 receptors is unaltered following carrageenan inflammation. AB - A descending facilitatory drive originating from superficial dorsal horn NK1 expressing neurones and relaying through parabrachial and rostroventral medial medulla to act on deep dorsal horn neurones, mediated through spinal 5HT3 receptors, was recently documented. To determine if this pathway plays a role in the pathophysiology of inflammation, we investigated the effects of spinally administered ondansetron (a selective 5HT3 receptor antagonist) on deep dorsal horn neuronal responses in carrageenan inflamed and naive animals using in vivo electrophysiology. The mechanical and thermal evoked responses of spinal neurones were dose dependently attenuated by ondansetron to a similar degree in both groups. In contrast, the electrically evoked responses (Abeta-, Adelta-, C-fibre evoked response and post-discharge) remained unaltered in both groups. Thus 5HT3 receptor mediated descending facilitation remains unaltered at this stage after tissue injury. PMID- 15135936 TI - Spinal neurons involved in the generation of at-level pain following spinal injury in the rat. AB - Using a conjugate of substance P and the ribosome-inactivating protein saporin, neurons expressing the neurokinin-1 receptor in lamina I of the spinal cord were targeted to determine their role in the expression of a spontaneous pain behavior following intraspinal injections of quisqualic acid in the rat. Treatment was carried out at the time of injury in order to prevent the onset of the behavior, and following onset in order to evaluate the potential clinical utility of this intervention. Treatment at the time of injury resulted in significant decreases in onset-time and severity of pain behavior, while treatment at the time of onset led to a significant reduction of the spontaneous self-directed behavior. The results suggest that the substrate for at-level pain following spinal cord injury includes a population of spinal neurons expressing the neurokinin-1 receptor in the superficial laminae of the spinal cord. PMID- 15135937 TI - Cerebellar neural responses related to actively and passively applied noxious thermal stimulation in human subjects: a parametric fMRI study. AB - Cerebellar activation is consistently found during noxious stimulation but little is known about its pain-related specificity. Under natural circumstances noxious stimuli are actively or passively delivered with concomitant tactile sensory stimulation. Using fMRI we therefore studied pain-related cerebellar activation with innocuous and noxious thermal stimuli in a parametric design taking motor execution as confounding factor into account. With respect to psychophysical pain ratings anterior vermal and ipsilateral hemispheric lobule VI activation was parametrically modulated for stimulus intensity in actively but not in passively elicited thermal stimulation. The cerebellum seems to be capable of distinguishing active from passive painful stimuli. PMID- 15135938 TI - Single-trial detection of human brain responses evoked by laser activation of Adelta-nociceptors using the wavelet transform of EEG epochs. AB - The aim of this study was to identify EEG changes induced by Adelta-nociceptor activation in single trials. In a preliminary experiment, intense CO(2) laser stimuli were delivered to the hand dorsum of five volunteers. The average amplitude of EEG epochs was estimated in the time-frequency (TF) domain using the continuous Morlet wavelet transform (CMT). The result was used as a TF filter enhancing Adelta-nociceptor induced EEG responses. In a second experiment, eight other subjects were delivered laser stimuli with six intensities. The CMT of each EEG epoch was computed. After applying the TF filter, amplitudes within a predefined interval were summed. Whether this sum predicted the occurrence of Adelta-nociceptor activation was tested using the reaction-time to discriminate between Adelta- or C-fibre mediated detection. Results showed that this method accurately identified single-trial EEG responses to Adelta-nociceptor activation. PMID- 15135939 TI - The involvement of the posterior cingulate gyrus in phasic pain processing of humans. AB - Cingulate cortex (CC) processing of phasic pain starts in caudal parts. This is evidenced by electrical brain source analysis (BESA(R)) and current reconstruction imaging (CURRY(R)) which identify the activated neuronal assemblies in the individual brain morphology, including the boundary element method for volume current calculation. Data were obtained from 30 subjects who experienced, in repeated sessions, infrared laser stimuli applied on temple, hand, and foot, or electrical shocks applied intracutaneously on finger tip. CC activity started around 230 ms (+/-20 ms; inter-subject SD.) and was localized in mid or posterior CC, in general in Brodmann's area 23 (intra-subject SD <5 mm; inter-subject SD <15 mm), independent of the kind and body site of noxious stimulation. PMID- 15135940 TI - Corticospinal excitability during painful self-stimulation in humans: a transcranial magnetic stimulation study. AB - We investigated changes in the corticospinal pattern of activity in healthy volunteers during sustained noxious and non-noxious mechanical stimulation of the first hand digit, resulting from active (self-stimulation) or passive (externally induced) pressing against a sharp or blunted tip. The results indicate that, in order to press a finger onto a noxious stimulus with the same force generated to press onto a non-noxious one, the motor cortex adopts a peculiar strategy in terms of recruitment of motor units. This is reflected by an increase of corticospinal excitability (as revealed by motor potentials evoked by transcranial magnetic stimulation of the contralateral primary motor cortex) and EMG activity of agonist muscles, possibly related to an increase of motor unit synchronization. PMID- 15135941 TI - Synaptic plasticity in the amygdala in a visceral pain model in rats. AB - The amygdala plays a key role in the emotional-affective component of pain. This study is the first to analyze synaptic plasticity in the central nucleus of the amygdala (CeA) in a model of visceral pain. Whole-cell patch-clamp recordings were made from neurons in the latero-capsular part of the CeA in brain slices from control rats and rats with zymosan-induced colitis (>6 h postinduction). Monosynaptic responses were evoked by electrical stimulation of afferents from the pontine parabrachial area (PB) and from the basolateral amygdala (BLA). Enhanced synaptic transmission was observed at the nociceptive PB-CeA synapse, but not at the polymodal BLA-CeA synapse, in rats with colitis. The frequency of action potentials evoked by direct current injection was increased in CeA neurons from colitis rats, suggesting enhanced neuronal excitability. Our results provide novel evidence for an important role of the CeA in visceral pain. PMID- 15135942 TI - Acupuncture and endorphins. AB - Acupuncture and electroacupuncture (EA) as complementary and alternative medicine have been accepted worldwide mainly for the treatment of acute and chronic pain. Studies on the mechanisms of action have revealed that endogenous opioid peptides in the central nervous system play an essential role in mediating the analgesic effect of EA. Further studies have shown that different kinds of neuropeptides are released by EA with different frequencies. For example, EA of 2 Hz accelerates the release of enkephalin, beta-endorphin and endomorphin, while that of 100 Hz selectively increases the release of dynorphin. A combination of the two frequencies produces a simultaneous release of all four opioid peptides, resulting in a maximal therapeutic effect. This finding has been verified in clinical studies in patients with various kinds of chronic pain including low back pain and diabetic neuropathic pain. PMID- 15135943 TI - Testosterone affects formalin-induced responses differently in male and female rats. AB - To evaluate the role of testosterone in pain modulation, we subcutaneously injected male and female rats with testosterone propionate (TP, 5 mg/kg in oil) or oil for 6 days; on the seventh day, all rats were subjected to the formalin test (10%, 50 microl). Behaviours were recorded in an open field (60 min). At the end of the formalin test, the rats were anaesthetized to collect blood from the abdominal aorta. Among the formalin-induced responses, licking was higher in females than males and was decreased by TP in females; jerking and flexing were only slightly affected by treatment. TP increased testosterone plasma levels in both sexes. These results indicate a role of testosterone in modulating formalin induced responses. The effects appear to be different in males and females. PMID- 15135944 TI - Alpha-2-adrenergic and opioid receptor additivity in rat locus coeruleus neurons. AB - Functional interactions between G protein-coupled receptors (GPCRs) are widely reported whereas the mechanisms underlying these interactions remain unclear. A rapidly growing literature suggests that many GPCRs form heteromeric complexes and that these complexes may possess pharmacological and functional properties that differ from their respective parent receptors. In order to investigate the functional relationship between co-localized GPCRs in intact, native CNS neurons, we used intracellular recording techniques in locus coeruleus (LC) slices. The LC consists of a homogenous population of cells, all of which co-express micro opioid and alpha(2A)-adrenergic receptors (microOR and alpha(2A)ARs, respectively). Co-administration of microOR and alpha(2A)AR agonists in vivo results in synergistic interactions in analgesic assays and heteromeric microOR alpha(2A)AR complexes have been detected in HEK-293 co-expressing the two receptors. The LC is therefore an excellent model system to explore these interactions. Dose-response and isobolographic analysis of agonist-induced hyperpolarization in individual LC neurons revealed that the interaction between these receptors was not synergistic but additive. This result suggests that co localized microOR and alpha(2)ARs are either not physically associated in these neurons or that heteromer formation may not fully explain the microOR-alpha(2A)AR synergistic interactions reported following agonist administration in vivo. PMID- 15135946 TI - An overview and analysis of site remediation technologies. AB - This paper presents an analysis of the site restoration techniques that may be employed in a variety of contaminated site cleanup programs. It is recognized that no single specific technology may be considered as a panacea for all contaminated site problems. An easy-to-use summary of the analysis of the important parameters that will help in the selection and implementation of one or more appropriate technologies in a defined set of site and contaminant characteristics is also included. PMID- 15135945 TI - Brain-derived neurotrophic factor-elicited or sciatic ligation-associated phosphorylation of cyclic AMP response element binding protein in the rat spinal dorsal horn is reduced by block of tyrosine kinase receptors. AB - Brain-derived neurotrophic factor (BDNF) and cyclic AMP response element binding protein (CREB) may critically contribute to injury-associated plasticity and thus to the development of persistent pain. In the present study we examined the potential interaction between CREB and BDNF in the spinal dorsal horn. Significant CREB phosphorylation was elicited by local application of BDNF (1 microg) onto the spinal dorsal horn of control, uninjured animals. The degree of phosphorylation was similar to that elicited by loose ligation of the sciatic nerve. The tyrosine kinase (Trk) blocker K252a (2 microg) significantly reduced the CREB phosphorylation elicited either by BDNF or the sciatic ligation. These data provided further support for the notion that at least some of the injury associated activation of CREB in the spinal dorsal horn may be dependent upon BDNF-mediated activation of Trk receptors. PMID- 15135947 TI - Environmental performance rating and disclosure: China's Green Watch program. AB - In this paper we describe a new incentive-based pollution control program in China, in which the environmental performance of firms is rated from best to worst using five colors--green, blue, yellow, red and black--and the ratings are disseminated to the public through the media. We focus on the first two municipal disclosure programs, which have been implemented at very different levels of economic and institutional development. In both cases, the increases in compliance with pollution regulations have been similar to increases produced by public disclosure programs in Indonesia, the Philippines, and Vietnam. The results suggest that incentives created by public disclosure may significantly reduce pollution in China, even though environmental NGO's play little role and there is no formal channel for public participation in environmental regulation. PMID- 15135948 TI - Promoting the use of environmental data collected by concerned citizens through information and communication technologies. AB - Public participation within environmental monitoring may contribute to increasing the knowledge on the state of the environment at the same time it promotes citizens' involvement in environmental protection. However, the use of voluntary collected data is limited due to a lack of confidence in data collection procedures. Additionally, data quality is often unknown and the data are usually dispersed and non-structured. Information and communication technologies (ICT) may promote the use of voluntary collected data through the development of a collaborative system that incorporates tools and methodologies to facilitate data collection, access and validation. Furthermore, the use of ICT may promote public involvement within environmental monitoring, since it facilitates communication among all the stakeholders. This paper analyses the role of ICT in developing a system for environmental collaborative monitoring intending to promote the use of volunteer collected data. It starts by analysing the role of volunteers within environmental monitoring and continues analysing the potential of ICT to take advantage of the benefits of using data collected by citizens. A collaborative system that allows the public to express its knowledge on the state of the environment is described. Special emphasis is given to tools that explore non traditional types of environmental data such as images, sounds and videos in association with spatial information. To illustrate the above mentioned concepts, a case study for beach quality monitoring developed within the Senses@Watch project, is described. PMID- 15135949 TI - Assessment of the effectiveness of scup bycatch-reduction regulations in the Loligo squid fishery. AB - Discard reduction is a component of the statutory requirements of the Sustainable Fisheries Act. One species of concern is scup, Stenotomus chrysops, discarded in the Loligo pealei fishery. Initially, regulations were imposed restricting the fishery in time and space to avoid areas and times associated with high scup discarding. Modified gear was required in 2003 on any boat fishing in areas otherwise closed to the fishery to reduce scup discarding. The purpose of this study was to evaluate the success of the 2003 net regulations and the potential influence of time-area closures (GRAs) in achieving a reduction in scup discarding. The regulations are based on three expectations. (1) Reduction in discarding in the Loligo squid fishery will materially reduce total scup discarding. (2) Exclusion of Loligo squid fishing vessels from the GRAs will result in these vessels fishing in areas that inherently produce fewer scup discards without equivalently increasing discarding in other sensitive species. (3) The use of a square-mesh large-mesh section in the extension will reduce scup discarding to the extent that otherwise would be achieved if the boats fished outside the GRAs without the economic cost imposed by redeploying the fleet. Analysis of the NMFS-NEFSC observer database offers no support for the belief that Expectation 1 has been met. Squid catches were too low to sustain a directed fishery in the northern GRA during this study. Thus, had this area been open, limited scup discarding would have occurred. In this study, squid catches averaged 1025 kg tow(-1) in the southern GRA. Thus, had the GRA been open, Loligo fishing would have taken place. Yet, in the 34 tows taken by two vessels, not a single scup was caught. Redeployment of the fleet clearly increased scup discarding in 2003. Thus, Expectation 2 was not met. Field tests demonstrated that the implemented net modification can produce reduced catches of mostly smaller-sized finfish without impairing squid catch, but the data also indicate that this result may not be routinely achieved. Thus, Expectation 3 was not completely met. Implementation of the 2003 net regulation was likely premature, in that the specification was not adequate to guarantee the desired results. The history of the scup discarding issue in the Loligo squid fishery demonstrates that discard reduction cannot be accomplished without adequate prior evaluation of the sources of discards, without the requisite and concomitant experimental evaluation of the results of regulatory reform, and without adequate commercial scale testing of prospective reforms prior to implementation. PMID- 15135950 TI - Natural materials for treatment of industrial effluents: comparative study of the retention of Cd, Zn and Co by calcite and hydroxyapatite. Part I: batch experiments. AB - This work explores the heavy metal retention capacity of materials developed from minerals that are abundant in nature, with low cost and minimum environmental impact. To accomplish this objective we have: (a) characterized commercial samples of calcite (CA) and hydroxyapatite (HAP)--including their surface properties (BET area, electrophoretic mobility, SEM, and X-ray energy dispersive spectroscopy); and, (b) qualified and quantified the interaction of Cd, Zn and Co with calcite (CaCO3) and hydroxyapatite [Ca5(PO4)3OH] through batch experiments, in a range of metal concentrations (4Zn>Co and Cd>Zn approximately Co, respectively. Retention increased with pCa and pH and could be modeled by: (a) a non-ideal ion exchange mechanism (Me/Ca) for the adsorption of Cd, Zn and Co onto CA; and, (b) a mechanism of non-ideal ion exchange and specific adsorption (Me/Ca and identical with PO4O-Me) in the case of HAP. The pH dependence is indirect in CA and is related to its solubility changes (pCa increases with pH, and so does sorption of Cd, Zn and Co). Both materials, HAP and CA, can be used for heavy metal retention. The former has better performance for water treatment due to its greater efficiency for the retention of Cd, Zn and Co (over two orders of magnitude per gram of material) and its lower solubility in a wide range of pH (6100 mM) and polyphosphate (0.01-0.5%) enhanced the antibacterial effects of the system in tomato juice and chicken skin extract, respectively. The findings indicate that the LPO system could probably be used to prevent the growth and survival of salmonellae in minimally processed fruit and vegetable products, but combination of the system with other preservatives or treatments would be needed to effectively inhibit growth and survival of salmonellae in animal products. PMID- 15135957 TI - Comparison between ultrafiltration and trichloroacetic acid precipitation method for concentration of Staphylococcus aureus enterotoxin in dairy samples. AB - Staphylococcus aureus enterotoxin that may be contained at low concentrations in milk and dairy products can cause food poisoning. To detect this enterotoxin at low concentrations, samples should be concentrated. We evaluated the performance of centrifugal ultrafiltration method (UF) in comparison with trichloroacetic acid precipitation method (TCA) for the concentration of S. aureus enterotoxin in milk and dairy products. S. aureus enterotoxin A (SEA) were added at various concentrations to ultra high-temperature heating process (UHT) milk, UHT concentrated skim milk, UHT skim milk powder, low heat-treated (LH) skim milk powder, and raw milk. SEA was concentrated by TCA and UF once a day on a total of 3 days by different researchers to prepare test solutions. The fluorescence value (TV) of test solutions was determined using an immunofluorescence autoanalyzer (miniVIDAS), and the linearity and slope of the regression line, relative standard deviation (RSD(RW)) at each added concentration, detection limit (DL), quantification limit (QL), and the recovery rate by each concentration method were obtained according to the guidelines of the International Conference on Harmonization (ICH). The slope of the regression line obtained by UF was steeper than that by TCA for all dairy samples excluding LH (74 degrees C, 20 s) skim milk powder. RSDRW, DL, and QL obtained by UF were comparable to or more excellent than those obtained by TCA. The procedure of UF was simpler than that of TCA. The recovery rate and rapidity were similar between the two methods. The DL and QL of enterotoxins other than SEA in dairy products by UF or TCA were estimated based on the DL and QL of SEA. In this estimation, consideration was given to reactions between each enterotoxin and its antibody, and also to the immunoactivity maintenance rate of each enterotoxin after addition of trichloroacetic acid in TCA. The estimated values were similar to those obtained by experiments using enterotoxin C1 (SEC1). UF using a centrifugal ultrafiltration membrane can be more readily performed and similar to or more reliable than TCA. UF combined with a miniVIDAS can be used for quantitative routine analysis. PMID- 15135958 TI - Partitioning of the variance in the growth parameters of Erwinia carotovora on vegetable products. AB - The objective of this paper was to estimate and partition the variability in the microbial growth model parameters describing the growth of Erwinia carotovora on pasteurised and non-pasteurised vegetable juice from laboratory experiments performed under different temperature-varying conditions. We partitioned the model parameter variance and covariance components into effects due to temperature profile and replicate using a maximum likelihood technique. Temperature profile and replicate were treated as random effects and the food substrate was treated as a fixed effect. The replicate variance component was small indicating a high level of control in this experiment. Our analysis of the combined E. carotovora growth data sets used the Baranyi primary microbial growth model along with the Ratkowsky secondary growth model. The variability in the microbial growth parameters estimated from these microbial growth experiments is essential for predicting the mean and variance through time of the E. carotovora population size in a product supply chain and is the basis for microbiological risk assessment and food product shelf-life estimation. The variance partitioning made here also assists in the management of optimal product distribution networks by identifying elements of the supply chain contributing most to product variability. PMID- 15135959 TI - Viability of lactic acid bacteria and bifidobacteria in fermented soymilk after drying, subsequent rehydration and storage. AB - To develop a probiotic dietary adjunct, soymilk fermented with various combinations of lactic acid bacteria (Streptococcus thermophilus and Lactobacillus acidophilus) and bifidobacteria (Bifidobacterium longum and Bifidobacterium infantis) was subjected to freeze-drying and spray-drying. Survival of the starter organisms during the drying process, subsequent rehydration at different temperatures and during a 4-month period of storage under different storage conditions was examined. After freeze-drying, lactic acid bacteria and bifidobacteria exhibited a survival percent of 46.2-75.1% and 43.2 51.9%, respectively, higher than that noted after spray-drying. Regardless of the drying condition, S. thermophilus showed a higher percentage of survival than L. acidophilus, while B. longum survived better than B. infantis. Further study with soymilk fermented with S. thermophilus and B. longum revealed that the freeze dried and spray-dried fermented soymilk rehydrated at 35-50 degrees C and 20 degrees C, respectively, was optimum for the recovery of the starter organisms. Both S. thermophilus and B. longum survived better in the freeze-dried than the spray-dried fermented soymilk during storage. A higher percent of survival was also noted for both the starter organisms when the dried fermented soymilk was stored at 4 degrees C than 25 degrees C. Holding the dried fermented soymilk in the laminated pouch enabled S. thermophilus and B. longum to exhibit a higher percentage of survival than in the deoxidant- and desiccant-containing glass or polyester (PET) bottle. Among all the packaging materials and storage temperatures tested, starter organisms were most stable in the dried fermented soymilk held in laminated pouch and stored at 4 degrees C. Under this storage condition, S. thermophilus and B. longum showed a survival percentage of 51.1% and 68.8%, respectively, in the freeze-dried fermented soymilk after 4 months of storage. Meanwhile, S. thermophilus and B. infantis in the spray-dried fermented soymilk showed a survival percent of 29.5% and 57.7%, respectively. PMID- 15135960 TI - Contribution of starter cultures to the proteolytic process of a fermented non dried whole muscle ham product. AB - Porcine longissimus dorsi muscles were cured by brine injection. Curing brine containing 15% (w/v) NaCl, 1.33% (w/v) glucose, 750 ppm sodium nitrite, and appropriate levels of either Lactobacillus sakei LAD, L. sakei LAD plus Kocuria varians FT4 (formally Micrococcus varians), L. sakei LAD plus papain and GDL (glucono-delta-lactone) plus K. varians FT4, was injected to the muscle at a pumping rate 15% w/v. The effect of these treatments on the proteolysis in the ham system was compared to a control ham, produced without starter culture and containing GDL acidulant to control pH and antibiotics to reduce the contribution of background microflora. Hydrolysis of sarcoplasmic and myofibrillar protein fractions was evaluated by SDS-PAGE and reverse phase-HPLC. Hams with different treatments were also investigated for differences in amino acid profile, protein and non-protein nitrogen level, colour, pH, water activity and moisture and microbiological evolution. There was no significant difference in the gross compositional analysis of any of the treatments compared to the control. There was no significant difference (p>0.05) in the protein content, non-protein nitrogen level, SDS-PAGE and free amino acid analysis between the control ham and ham inoculated with proteolytic starter culture. However, it was observed that hams containing starter cultures exhibited decreases in certain peptide fractions and corresponding increases in some free amino acids compared to the uninoculated control. It can be concluded that, while the principle mechanisms resulting in the proteolysis of this non-dried ham product involve the activity of endogeneous cathepsins, the addition of proteolytic starter cultures influence the amino acid profile thereby potentially enhancing the sensorial attributes of the ham. PMID- 15135961 TI - A quantitative risk assessment model for Salmonella and whole chickens. AB - Existing data and predictive models were used to define the input settings of a previously developed but modified quantitative risk assessment model (QRAM) for Salmonella and whole chickens. The QRAM was constructed in an Excel spreadsheet and was simulated using @Risk. The retail-to-table pathway was modeled as a series of unit operations and associated pathogen events that included initial contamination at retail, growth during consumer transport, thermal inactivation during cooking, cross-contamination during serving, and dose response after consumption. Published data as well as predictive models for growth and thermal inactivation of Salmonella were used to establish input settings. Noncontaminated chickens were simulated so that the QRAM could predict changes in the incidence of Salmonella contamination. The incidence of Salmonella contamination changed from 30% at retail to 0.16% after cooking to 4% at consumption. Salmonella growth on chickens during consumer transport was the only pathogen event that did not impact the risk of salmonellosis. For the scenario simulated, the QRAM predicted 0.44 cases of salmonellosis per 100,000 consumers, which was consistent with recent epidemiological data that indicate a rate of 0.66-0.88 cases of salmonellosis per 100,000 consumers of chicken. Although the QRAM was in agreement with the epidemiological data, surrogate data and models were used, assumptions were made, and potentially important unit operations and pathogen events were not included because of data gaps and thus, further refinement of the QRAM is needed. PMID- 15135962 TI - Determinations of the antimutagenic activities of several probiotic bifidobacteria under acidic and bile conditions against benzo[a]pyrene by a modified Ames test. AB - Antimutagenic activities of six bifidobacteria, after acidic and bile treatment mimicking gastrointestinal conditions, against benzo[a]pyrene (B[a]P) were determined by a modified Ames test using Salmonella typhimurium TA 100. Results showed that when bifidobacteria were treated at pH 2.0 for 3 h or 1% bile for 6 h, their antimutagenic activities against B[a]P were increased as compared to controls at pH 7.0 for 0 h. After sequential acidic pH and bile treatments, Bifidobacterium lactis Bb-12 displayed the highest antimutagenic activity (71.5%), although its viable cell number was less than 2.0 log cfu/ml. The antimutagenic activity of B. lactis Bb-12 against B[a]P was increased as pH values were increased from 2.0 to 7.0, and reaction time was extended from 1 to 3 h. However, antimutagenic activity was decreased as bile salt concentration was increased from 0.5% to 2.0%. The antimutagenic activity of B. lactis Bb-12 against B[a]P was increased in the presence of whole milk (WM), semiskimmed milk (SSM) and skimmed milk (SM). When B. lactis Bb-12 was preincubated with B[a]P and milk substrates, its antimutagenic activity was increased to 99-100%. PMID- 15135963 TI - Quantitative contamination and transfer of Escherichia coli from foods by houseflies, Musca domestica L. (Diptera: Muscidae). AB - The housefly, Musca domestica L. (Diptera: Muscidae), is recognized as an important factor in the dissemination of various infectious diseases such as cholera, shigellosis, and salmonellosis. They can also serve as a cross contamination vector for other foodborne pathogens. However, the potential for bacterial transfer by houseflies has been demonstrated in a qualitative rather than quantitative manner. In this study, the numbers of bacteria a housefly can carry on its body and transfer to a clean surface after exposure to a sugar-milk aqueous solution, steak, and potato salad contaminated with a fluorescent gene Escherichia coli (8 log10 CFU/ml) were determined. In the first series of experiments to quantify bacterial numbers on the flies, about 40-60 flies were transferred into a sterile cage, exposed to the food for 30 min, the flies immobilized and the attached E. coli on each fly enumerated. Detectable E. coli (>1.7 log10 CFU/fly) were found on 43% (29/67), 53% (23/43), and 62% (32/52) of the flies in the cages with sugar/milk, steak, and potato salad, respectively. For the positive flies, the geometric mean carriage (log10 CFU/fly) was 2.93+/ 1.24 for sugar-milk, 3.77+/-1.28 for steak, and 2.25+/-0.64 for the potato salad. In the second series of experiments, the transfer of bacteria by individual flies from contaminated food to the inner surface of a sterile jar per each landing was determined. E. coli transferred from the sugar-milk was 3.5+/-0.7 log10 CFU/fly landing, 3.9+/-0.7 for steak and 2.61+/-1.16 for the potato salad. From the initial contamination levels of bacteria and the number of transferred bacteria, it can be calculated that flies contaminate clean surfaces with approximately 0.1 mg of food per landing. PMID- 15135964 TI - Differential acquisition of food-reinforced disk pressing by CD-1, BALB/cJ and C57BL/6J mice. AB - Developments in the genetic manipulation of mice have intensified interest in the relation between genes, environment and learned behavior, which in turn has led to exploration of experimental procedures for assessing genetic influences on learning using methods such as response acquisition. The requirement for multiple experimental control groups in such genetic comparisons studies amplifies the need for reliable, instrument-directed, learning assessment paradigms. The purpose of the present experiment was to implement such a procedure in several successive phases, including pre-food basal rates of interaction with sensors in a chamber, food-hopper training, and a simple disk-baiting procedure that produced differentially food-reinforced disk pressing (FR1) and provided quantitative measures of learning in outbred (CD-1) and inbred (C57BL/6J, BALB/cJ) mice. Response acquisition performances were measured in terms of the number of correct food-producing responses (disk press to hopper entry in less than 5s). The three mouse types showed differences in several performance measures prior to response acquisition training. Pre-food basal performances did not predict subsequent rate of acquisition of the target response. On average, CD 1 mice met a criterion of 50 food-producing responses slightly, but not significantly, faster than BALB/cJ mice. The C57BL/6J mice took significantly longer to meet the learning criterion and had slower response rates, due to longer after-reinforcement pausing. Procedural differences (massed versus partial/distributed training) and reinforcement parameters (duration of access) may differentially affect different mouse types independently of genetic differences in response acquisition. PMID- 15135965 TI - Sniffing human sex-steroid derived compounds modulates mood, memory and autonomic nervous system function in specific behavioral contexts. AB - We asked whether the effects of exposure to two human sex-steroid derived compounds were context dependent. The effects of sniffing 4,16-androstadien-3-one (AND) and 1,3,5(10),16-estratetraen-3-ol (EST) on mood, memory, and autonomic nervous system responses were explored in 72 participants. Subjects were tested with AND, EST, or a Control compound within four mood contexts: neutral, sexually aroused, sad and happy. These moods were successfully induced using selected film segments (P < 0.0001). During the neutral context, none of the compounds affected mood or autonomic nervous system function. However, compound effects were significantly increased within arousing contexts. During the sexually arousing context, both compounds increased sexual arousal (P < 0.029). During the sad context, AND maintained positive mood in women (P< 0.050) and increased negative mood in men (P < 0.031). Memory for events during the sad context was impaired by AND in women (P < 0.047) but not in men. Finally, effects of AND on physiology were observed during the sexually arousing context whereby AND increased skin temperature in both sexes (P < 0.022) but reduced abdominal respiration rate in men only (P < 0.034). These results suggest that sex-steroidal compounds modulate mood, memory and autonomic nervous system responses and increase their significance within specific behavioral contexts. These findings lend support to a specific role for these compounds in chemical communication between humans. PMID- 15135966 TI - Electrolytic lesions of the ventral subiculum weakly alter spatial memory but potentiate amphetamine-induced locomotion. AB - Adult Long-Evans male rats were subjected to electrolytic lesions of the ventral subiculum, and tested for locomotor activity in the home cage, reference and working memory in the water maze, working memory in the radial maze, and D amphetamine-induced locomotion (1mg/kg, i.p.). When compared to their sham operated counterparts, lesioned rats showed nocturnal hyperactivity, no reference memory deficit, but working memory was impaired in the water maze and during the initial stage of radial-maze testing. Their locomotor responsiveness to D amphetamine was exaggerated. Histological verifications confirmed lesions in the ventral subiculum. Material stained for acetylcholinesterase activity indicated septohippocampal and commissural/associational sprouting, accounting for partial damage to the perforant paths. These results showed that ventral subiculum lesions (i) do not alter the capability of rats to learn repeatedly presented spatial information, and (ii) impair, but do not prevent, spatial working memory, suggesting that the ventral subiculum is preferentially involved in short-term memory for spatial locations. Given the electrolytic nature of the lesion, the lesion-induced potentiation of the locomotor response to amphetamine is probably easier explained by partial disruption of the perforant paths than by damage to neurons of the ventral subiculum. PMID- 15135967 TI - Survey of embryonic stem cell line source strains in the water maze reveals superior reversal learning of 129S6/SvEvTac mice. AB - The availability of pluripotent embryonic stem (ES) cells for gene targeting has resulted in laboratory mice becoming important animal models of human neurological disease. Inbred strains of mice differ in many behavioural phenotypes, such that the same gene mutation can appear to have different phenotypic effects when introduced onto different genetic backgrounds. Prior knowledge of the behavioural phenotypes of the inbred strains used for gene targeting would, therefore, allow the selection of the most appropriate genetic background for the hypothesis to be tested. With this in mind, we tested eight strains of mice (129S1/SvImJ, 129S2/SvPasIcoCrlBR, 129S6/SvEvTac, B6129SF1/J, C57BL/6J, C57BL/6N, LP/J and SM/J), including the sources of five ES cell lines commonly used for gene targeting, in the spatial (submerged platform) version of the Morris water maze, the most widely used paradigm to evaluate the cognitive abilities of genetically modified mice. The three 129 substrain sources of ES cell lines demonstrated spatial learning in the water maze that was superior to that of C57BL/6J, the inbred strain most commonly used for the maintenance and phenotypic testing of mutations. In addition, 129S6/SvEvTac was unique amongst the eight strains tested in having a particular capacity for reversal learning, when the submerged platform was relocated to the opposite quadrant. We conclude that some substrains of 129 could provide suitable genetic backgrounds for testing gene mutations that might be expected to impair cognitive function, thus negating the need to backcross to C57BL/6J, thereby avoiding the so-called "flanking gene problem". PMID- 15135968 TI - Decreased brain docosahexaenoic acid during development alters dopamine-related behaviors in adult rats that are differentially affected by dietary remediation. AB - Docosahexaenoic acid (DHA) is a major component of neuronal membranes. In rats, low brain levels of DHA during development produce alterations in the mesocortical and mesolimbic dopamine systems. In this study, male Long-Evans rats (n = 6-7 per group) were raised from conception on diets with (control) or without alpha-linolenic acid, the dietary precursor of DHA. The deficient diet reduced brain DHA to 80% of that of control animals at maturity. At weaning (P21), half of the pups from each deficient litter were changed to a remediation diet enriched in DHA by the addition of fish oil. Beginning at P56, rats were subjected to a battery of behavioral tests. Catalepsy induced by haloperidol (3mg/kg, i.p.) in deficient animals was 49% of that observed in control animals (P < 0.05). Catalepsy in remediated animals was similar to that observed for control animals. In a test of locomotor activity in a novel environment, deficient animals exhibited 187% of the activity of control animals (P < 0.05). Locomotor activity stimulated by d-amphetamine (0.5mg/kg, s.c.) in deficient animals was 145% of that observed in control animals (P < 0.05). Basal and amphetamine-stimulated activity of remediated animals was similar to that of deficient animals. Response to a thermal stimulus was not different between groups. These findings demonstrate that rats exposed from conception to a diet that produces a relatively modest decrease in brain DHA content exhibit alterations in adult behavior indicative of altered dopaminergic function. Some of these behavioral alterations were reversed by dietary remediation initiated at weaning. PMID- 15135969 TI - Dissociation of core and shell single-unit activity in the nucleus accumbens in free-choice novelty. AB - Core and shell regions of the nucleus accumbens (NAcc) have been implicated in different aspects of goal-directed behavior. To assess these differences at the neuronal level, we evaluated core and shell single-unit activity in eight rats during one-trial, free-choice entry into a novel compartment. Changes in firing rate during approach of (orientation toward) and entry into (nose cross) novelty were assessed relative to a pre-novelty baseline when the animals were behaviorally active. Increases and decreases in neuronal activity were recorded in both regions during both phases of the novelty response. The regional distribution of these responses, however, was significantly different during the approach phase with roughly equal proportions of neuronal excitations (8/29) and inhibitions (6/29) in core but a shift away from excitation (2/40) toward inhibition (12/40) in shell. No regional differences emerged during subsequent approach of the familiar compartment from the novel chamber or during entry to either the novel or familiar compartment. Taken together, our results during approach to free-choice novelty indicate a regional dissociation in NAcc firing rate may play a role in appetitive behavior. PMID- 15135970 TI - Infralimbic D1 receptor agonist effects on spontaneous novelty exploration and anxiety-like defensive responding in CD-1 mice. AB - Mesocortical dopamine (DA) terminals in the ventromedial prefrontal cortex (vmPFC) integrate cognitive/emotional processing functions underlying adaptive and appropriate behavioral responding to stressful environmental events. Results from several studies have also shown that stressor-enhanced prefrontal DA activation exerts detrimental effects on cognitive performance. However, questions have arisen as to whether stressor-enhanced vmPFC DA transmission exerts direct control over conditioned or unconditioned responses to threatening events, or whether enhanced prefrontal DA transmission gates cognitive processing to facilitate adaptive responding in threatening situations. We have previously shown that infralimbic (IL) vmPFC dopamine D2 agonist and antagonist drug infusions reduced anxiety-like responding in the elevated plus-maze (EPM) and disrupted spontaneous exploration in the Y-maze in CD-1 mice. In the present study, the effects of IL vmPFC infusions of the specific D1 receptor agonist, SKF 81297, in CD-1 mice were evaluated on spontaneous exploration in the Y-maze, anxiety-like responding in a 2-trial elevated plus-maze procedure, and anti predator defensive responding in the Mouse Defense Test Battery (MDTB). SKF-81297 infusions disrupted spontaneous alternation performance along with potentiated repetitive 2-arm responding in the Y-maze. In the elevated plus-maze, pre-trial 1 IL SKF-81297 infusions reduced anxiety-like responding (enhanced open arm entries and time ratio, unprotected stretch attends and head dips), and reduced closed arm time ratio and protected risk assessment activity (protected stretch attends). In trial 2, 24h later (no drug infusions), open arm entries, open time ratio, and unprotected head dips remained enhanced relative to trial 2 vehicle controls. In the MDTB, avoidance distance was enhanced in the approach test; risk assessment (approach) was enhanced in the closed alley test; and defensive threat (upright postures) was enhanced in the forced contact test. Results are discussed with respect to possible influences of IL vmPFC DA receptors on cognitively mediated responding to differing levels of threat in mice. PMID- 15135972 TI - Removal of the accessory olfactory bulbs promotes maternal behavior in virgin rabbits. AB - Virgin rabbits exposed to foster pups for 14 days did not show maternal responsiveness. However, surgical removal of the accessory olfactory bulbs (AOB) activated maternal responsiveness (crouching over the litter inside the nest box for about 3min, which is the normal duration of a nursing bout) in 37% of virgin rabbits (P < 0.007). This behavior appeared abruptly and was first observed between days 3 and 13 of pup exposure. This variation in the latency to respond maternally was not related with the number of sniffings or entrances into the nest box displayed by a female on the days that preceded crouching over the litter. Maternal responsiveness was not observed in any AOB-lesioned animals that were also ovariectomized (P < 0.02 versus AOB-lesioned with ovaries). These results indicate a tonic inhibitory action of the AOB over the expression of maternal behavior in virgin rabbits and a stimulation of maternal responsiveness by ovarian hormones following AOB lesions. PMID- 15135971 TI - The effect of reversible inactivation of the supramammillary nucleus on passive avoidance learning in rats. AB - Previous studies have shown that the presence of hippocampal theta activity is important for learning and memory, and that the medial supramammillary nucleus (mSuM) is involved in the control of the frequency of theta rhythm. It has also been shown that the depression of mSuM activity by chlordiazepoxide causes modest impairment of spatial learning. On the other hand, the lateral supramammillary nucleus (lSum) increases long-term potentiation (LTP) of hippocampal population spikes. However, to our knowledge, no reports exist concerning the role of the supramammillary area (SuM) in passive avoidance (PA) learning. In the present study, rats were chronically implanted with a cannula aimed at SuM and were trained on a step-through PA task. They received intra-SuM injection of lidocaine or saline at the following intervals: 5 min before training, 5, 90, and 360 min after the acquisition trial, or 5 min before the retrieval test. When lidocaine was injected 5 min before training there was no effect on acquisition of PA but retrieval was significantly poorer than the control group injected with saline. Lidocaine injection 5 min after the acquisition trial impaired PA retention, but reversible inactivation of SuM at 90 and 360 min after training and 5 min before the retrieval test showed no significant effect on PA retention. It can be concluded that SuM contributes to PA consolidation at least 5 min after the acquisition trial and that this effect may be accomplished through SuM projections to the septal and/or hippocampal areas participating in the PA memorization processes. PMID- 15135973 TI - Spatio-temporal brain activation profiles associated with line bisection judgments and double simultaneous visual stimulation. AB - We used magnetic source imaging (MSI) to investigate the spatio-temporal patterns of brain activity associated with line bisection judgments and double simultaneous visual stimulation in 14 healthy adults. Consistent with lesion and hemodynamic neuroimaging studies, we found the greatest number of activity sources in right inferior parietal cortex. These sources were most prominent, on average, between 200 and 300 ms after the onset of single (left, right, or center) target stimuli. A greater number of significant activity sources were found in right inferior parietal, occipital, and prefrontal cortices during bilateral compared with unilateral stimulus presentation. Based on these observations, we suggest that a more parsimonious physiological explanation of visual extinction than the hemispheric rivalry account may be the additional neuronal excitation required in right occipital and parietal cortices for accurate bilateral visual perception. PMID- 15135974 TI - Amygdala c-Fos induction corresponds to unconditioned and conditioned aversive stimuli but not to freezing. AB - These experiments examined the relationship between freezing and c-Fos expression in the amygdala. In Experiment 1 freezing was elevated during a period immediately following shock in rats that remained in the shock context, but not in rats that were moved to a different, neutral context. The two groups showed equally elevated c-Fos levels in both the central (CeA) and lateral (LA) nuclei. In Experiment 2 rats were shocked in one compartment (paired) and not shocked in another, distinct compartment (unpaired). Rats re-exposed to the paired compartment 24h later froze more than rats exposed to the unpaired compartment, and rats in both groups froze more than un-shocked rats. c-Fos protein expression in CeA, LA and basolateral (BLA) nucleus was elevated in the rats exposed to the paired compartment but not in rats exposed to the unpaired compartment. Thus, c Fos expression was induced by exposure to both unconditioned and conditioned stimuli, although it is unclear if the same cell population was activated in both cases. Neither case of c-Fos expression coincided with the occurrence of freezing. c-Fos expression may represent neural activity in LA and CeA produced by exposure to unconditioned cues and activity in BLA, LA and CeA produced by conditioned cues. This activity may contribute to an aversive affective state (or "fear"). Behaviors promoted by this state, such as freezing, may be mediated in other brain areas, or by other neurons in the amygdala. PMID- 15135975 TI - An animal model of antipsychotic-induced weight gain. AB - We have established an animal model for olanzapine-induced body weight gain, and used it to explore the relation between this weight gain, excessive food consumption, gross motor activity, and macronutrient choice. Female Sprague Dawley rats received olanzapine (OLAN) or diluent (1.2mg/kg per day) via gavage for 10 days. Rats receiving OLAN exhibited significant increases in body weight when compared with control rats. Body weight returned to control levels once OLAN treatment was discontinued. Food consumption among the OLAN-treated group was significantly greater than among control rats between 6 and 10 days of treatment. Between 4 and 10 days of treatment, feed efficiency (grams of weight gained/grams of food consumed) was also significantly greater among animals receiving OLAN. In contrast, chronic administration of haloperidol (0.04mg/kg; q.d.; gavage) did not influence body weight or food consumption of treated rats. Gross motor activity was significantly reduced by OLAN between 1 and 10 days of treatment, also returning to control levels when treatment was discontinued. No significant changes were observed in brain DA, DOPAC, HVA or 5-HIAA among animals receiving OLAN daily for 30 days; however, 5-HT levels were significantly elevated. In contrast, acute (1.2mg/kg; 2h; i.p.) administration of OLAN significantly increased brain DOPAC and HVA levels without affecting those of 5-HT or 5-HIAA. OLAN (1.2mg/kg; q.d.; 10 days) administration did not alter macronutrient choice (carbohydrate:protein ratio) of rats. These data show that an animal model of OLAN-induced weight gain is readily generated, and suggest that the weight gain results at least in part from increased food intake, reduced gross motor activity, and enhanced feed efficiency. PMID- 15135976 TI - Effect of estrogen on the lordosis-inhibiting action of ketanserin and SB 206553. AB - The effects of the 5-HT(2A/2C) receptor antagonist, ketanserin, and the 5-HT(2C) receptor antagonist, SB 206553, on lordosis behavior were investigated in ovariectomized rats hormonally primed with estradiol benzoate (EB) (0.5 or 25 microg) and progesterone (500 microg). Both ketanserin and SB 206553 inhibited lordosis behavior after infusion into the ventromedial nucleus of the hypothalamus (VMN), but ketanserin was slightly more effective than the 5-HT(2C) receptor antagonist. Either drug was more effective in rats primed with 0.5 microg EB than in rats hormonally primed with 25 microg EB. These findings support the suggestion that estrogen may enhance functioning of the 5-HT(2) receptor family and thereby protect against the 5-HT(2) receptor antagonists. These data are consistent with prior suggestions that estrogen modulates functioning of 5-HT(2) receptors within the VMN and that 5-HT(2) receptors play a facilitatory role in the modulation of female rat lordosis behavior. PMID- 15135977 TI - Attentional processes and cognitive performance during expectancy of painful galvanic stimulations: a high-resolution EEG study. AB - In the present high-resolution electroencephalographic (EEG) study, an omitted stimulus paradigm induced a strong expectancy for a predictable painful stimulation (nonpainful in the control condition). During the expectancy of pain, concurrent cognitive demands were superimposed. The aim was to investigate the effects on primary sensorimotor and central midline areas of the competition among concurrent attentional processes related to cognition and pain expectancy, as indexed by behavioral performance and EEG data. A main issue was whether cognitive performance decreases, due to a re-allocation of attentional resources on primary sensorimotor and midline areas for the anticipation of pain. Behavioral results showed no differences in the cognitive (working memory) performance during the expectancy of nonpainful versus painful stimulations. In parallel, anticipatory event-related potentials (ERPs) were negligible in line with a low emotional reactivity/alertness as revealed by heart rate deceleration (HRD), skin conductance response (SCR), and low-band (6-10Hz) alpha EEG oscillations. In contrast, high-band alpha EEG oscillations (10-12Hz) over the contralateral primary sensorimotor cortex decreased more during the expectancy of painful compared to nonpainful stimuli, in line with an increased anticipatory preparation of the somatosensory channel. These findings provide further evidence on the fact that attentional processes at the basis of cognition can be defended by the anticipation of pain, at least when the incoming painful stimuli are repetitive and predictable. This happens even if the brain increases preparatory processes of the specific sensory channel to be targeted by the painful stimulus. PMID- 15135978 TI - Environmental modulation of the interoceptive effects of amphetamine in the rat. AB - The aim of the present study was to investigate the effect of environmental novelty on amphetamine discrimination. Two groups of rats (home group and novelty group) were trained to perform water-reinforced operant behavior in cages equipped with two levers and a retractable liquid dipper (0.1cc cup). The experimental procedures for the two groups were identical except for the fact that home rats were housed and tested in the operant cages whereas novelty rats were transferred daily to these cages for the testing sessions (25min). The rats were trained to discriminate one of two doses of amphetamine (0.25 and 0.5mg/kg) from saline. Saline and amphetamine were administered intraperitoneally immediately before each daily session according to a semi-random schedule for a total of 50 sessions. Reinforcements were delivered according to a fixed ratio that was increased daily from 2 to 30. Successful drug discrimination was achieved when activity on the appropriate lever before the first reinforcement was greater than 80% of total activity, on at least seven out of eight consecutive sessions. When training was conducted with 0.25mg/kg of amphetamine, no home rat acquired drug discrimination whereas 56% of novelty rats reached criterion. When training was conducted with 0.5mg/kg of amphetamine, drug discrimination was achieved by 50% of home rat versus 86% of novelty rats. These findings indicate that environmental novelty can alter the ability of amphetamine to produce interoceptive cues. PMID- 15135979 TI - Individual differences in sensitivity to factors provoking reinstatement of cocaine-seeking behavior. AB - Among cocaine addicts, there is a strong variation in response to relapse provoking factors like conditioned cues and renewed contact with the drug. Here we show that such large individual differences also exist in rats. Reinstatement of extinguished cocaine-seeking behavior was triggered by contingent presentation of a cocaine-conditioned cue or an amphetamine priming injection. We found no positive correlation between cue- and drug-controlled reinstatement of cocaine seeking. Rather, a slight, but significant negative correlation was observed, which was particularly evident in two subgroups of rats that responded highest following either amphetamine priming or cue presentation. A large middle group responded equally for both relapse provoking factors. Further, cocaine-seeking behavior during the first extinction session correlated positively with cue induced reinstatement. In conclusion, the present findings indicate that the therapeutic efficacy of relapse prevention strategies may depend on individual sensitivity to distinct relapse provoking stimuli. PMID- 15135980 TI - Sub-clinical infection as an effective protocol for obtaining anti-Leishmania chagasi amastigote antibodies of different animal species. AB - This work aims at identifying an effective protocol to raise anti-Leishmania chagasi amastigote antibodies in different animal species. Protocols of immunization by subcutaneous injections of L. chagasi promastigote and amastigote lysates or by either intravenous or subcutaneous inoculation of live metacyclic promastigotes were assessed in mice, rabbits, and dogs. The immunization with live promastigotes produced a strong humoral immune response against L. chagasi amastigotes in all three animal species. The sera from animals immunized with the promastigote lysate did not react with amastigotes and, conversely, the sera from mice immunized with the amastigote lysate did not react with promastigotes. Taken all data together, the immunization through infection with metacyclic promastigotes was considered the most satisfactory way to immunize animals for obtaining anti-amastigote and anti-promastigote antibodies, since it did not only allowed the obtention of antibody against the two forms of the parasite, but it is also cheap, less laborious than carrying out the purification of amastigotes from infected tissues and avoid the use of a large number of hamsters for obtention the amastigotes, necessary to produce the immunogenic lysates. Furthermore, this immunization protocol was comparable to the amastigote lysate immunization protocol for the obtaining of mouse monoclonal antibodies (mAbs). PMID- 15135981 TI - Characterisation of the syncytia formed by VHS salmonid rhabdovirus G gene transfected cells. AB - Protein G expression and cell-to-cell fusion of cells transfected with the G gene of viral haemorrhagic septicaemia virus (VHSV) has been characterised. The presence of protein G in the membrane of transfected cells was confirmed by staining with Abs (Abs) and FACS. The subsequent formation of syncytia by membrane fusion of transfected cells required transfection with a wild type G gene and a low pH step. Mice Abs made against the protein G regions involved in fusion and neutralising monoclonal Abs (MAbs) as well as MAbs against some linear epitopes inhibited syncytia formation, thus confirming that syncytia formation was G-dependent. Similarly, Abs from trout immunised with purified VHSV or protein G inhibited syncytia formation whereas Abs from non-immunised or non infected animals did not. Abs from mice or trout with the highest neutralisation titres also showed the highest percentage of inhibition of syncytia. While the main utility of these observations might be to further the understanding of the complex trout antibody response against VHSV and in the follow up of VHSV immunisation attempts, they may also have some future diagnostic potential for countries were work with VHSV is not allowed. PMID- 15135982 TI - The biological effects of five feline IFN-alpha subtypes. AB - IFN-alpha has been shown to induce both antiviral and antiproliferative activities in animals. This report describes the biological activity of five recently identified feline IFN-alpha subtypes expressed in the Chinese hamster ovary (CHO) cell line (rfeIFN-alpha1[CHO], rfeIFN-alpha2[CHO], rfeIFN alpha3[CHO], rfeIFN-alpha5[CHO] and rfeIFN-alpha6[CHO]) and the feIFN-alpha6 subtype expressed in and purified from Pichia pastoris (rfeIFN-alpha6[P. pastoris]). The rfeIFN-alpha[CHO] subtypes were tested for antiviral activity against either Vesicular stomatitis virus (VSV) or feline calicivirus (FCV) infected feline embryonic fibroblast cell line (AH927) or Crandell feline kidney cell line (CRFK). Antiviral activity was induced against both VSV and FCV infected AH927 cells and VSV infected CRFK cells by all five of the rfeIFN alpha[CHO] subtypes and rfeIFN-alpha6[P. pastoris]. In addition, the IFN-alpha inducible Mx gene (associated with antiviral activity) was upregulated in vivo 24 h following treatment with rfeIFN-alpha6[P. pastoris], compared to baseline levels seen prior to treatment. All of the rfeIFN-alpha[CHO] subtypes and rfeIFN alpha6[P. pastoris] exhibited antiproliferative activity in the FeT-J cell line (an IL-2 independent feline T-cell line). Both necrosis and apoptosis were observed in rfeIFN-alpha6[P. pastoris]-treated FeT-J cells. The rfeIFN alpha3[CHO] subtype consistently exhibited lower antiviral and antiproliferative activity compared to that observed with the other four rfeIFN-alpha[CHO] subtypes. In summary, this paper demonstrates that five previously described feIFN-alpha subtypes induce both antiviral and antiproliferative activities in vitro and are capable of upregulating the feMx gene in vivo. PMID- 15135983 TI - In utero infection with PRRS virus modulates cellular functions of blood monocytes and alveolar lung macrophages in piglets. AB - The putative immunosuppressive effect of PRRS virus (PRRSV) on innate immune responses was studied in piglets infected in utero with PRRSV. Phagocytosis and oxidative burst capacities in 2-, 4- and 6-week-old in utero infected piglets were investigated and compared with age-matched control piglets. Phagocytic capacity of blood monocytes against Salmonella bacteria was investigated by flow cytometry. Oxidative burst in blood monocytes and in alveolar lung macrophages was investigated by luminol- and lucigenin-enhanced chemiluminescence, respectively. Decreased phagocytosis against Salmonella was found in blood monocytes from 4- and 6-week-old infected piglets compared to controls. In contrast, 2-week-old infected piglets showed phagocytic responses comparable to age matched control piglets. While oxidative burst capacity was increased in blood (PBMC) from in utero PRRSV infected piglets, the oxidative burst capacity of alveolar lung macrophages was decreased, especially in 2- and 4-week-old piglets, compared to age-matched control piglets. The present results indicate that in utero infection with PRRSV inhibits phagocytosis against Salmonella in blood monocytes as well as the oxidative burst capacity of alveolar macrophages. These observations indicate that PRRSV in utero infection induces at state of immunosuppression in piglets paving the way for enhanced secondary infections. PMID- 15135984 TI - The immunopathogenesis of flea allergy dermatitis in dogs, an experimental study. AB - In this study, we investigated the development of clinical disease and immune responses in the development of an experimental model of flea allergy dermatitis. Dogs were randomly divided into four treatment groups and were infested with fleas on two different feeding schedules (continuous and episodic). Group 1 consisted of four non-exposed dogs (negative controls) and Group 2 consisted of six dogs exposed to fleas continually. Groups 3 and 4 consisted of 14 dogs each that were exposed to fleas on an episodic schedule (two consecutive days every other week for 12 weeks). Group 4 also received intraperitoneal injections of a low dose of lectin (ricin) with immunomodulatory properties. The purpose of Group 4 was to investigate the effects of ricin on enhancing the development of clinical signs, flea antigen-specific IgE levels and altering the number of CD4+ and CD8+ T cell subsets in peripheral blood. Clinical signs developed in all flea exposed dogs, however, the dermatology lesion scores were less and shorter in duration for continuously exposed dogs compared to episodic exposed dogs, independent of ricin treatment. Lesion development was concentrated in the flea triangle and consisted principally of erythema, followed by alopecia, excoriation, papules, and crusts. CD4+ and CD8+ lymphocyte subsets or IgE levels were not altered by ricin treatment. Flea antigen-specific IgE values were highest in dogs exposed to fleas on a continuous basis compared to those episodically exposed. A greater percentage of clinical responder dogs with negative flea-specific IgE titers or negative intradermal test (IDT) were present in the episodic exposure groups than in the continuous exposure group. IgE titers corresponded slightly better with clinical responders than the IDT. The agreement between the IgE titers and IDT was good (weighted K = 0.67). Histopathology of skin samples were consistent with a Type I hypersensitivity. In conclusion, we were able to develop a model of flea allergy dermatitis by experimentally exposing dogs to fleas on an episodic and continuous feeding schedule. In this study, continuously exposed dogs did not develop immunotolerance, and ricin did not enhance the development of FAD. PMID- 15135985 TI - BHV-1 infection and inflammatory cytokines amplify the interaction of Mannheimia haemolytica leukotoxin with bovine peripheral blood mononuclear cells in vitro. AB - Bovine herpesvirus-1 (BHV-1) has been reported to increase the susceptibility of cattle to respiratory disease caused by Mannheimia (Pasteurella) haemolytica A1. The principal virulence factor of M. haemolytica is a leukotoxin (LKT) that can specifically kill ruminant leukocytes following its binding to the beta2-integrin CD11a/CD18 (lymphocyte function-associated antigen 1 (LFA-1)). In this study, we investigated the effects of experimental infection of bovine peripheral blood mononuclear cells (MNCs) with BHV-1 in vitro, on the subsequent interaction of these cells with the M. haemolytica LKT. We found that BHV-1 infection increased LFA-1 expression (as assessed by flow cytometry), and subsequently enhanced LKT binding and cytotoxicity to bovine MNCs. We also found that BHV-1 infection increased CD18, IL-1beta, and IFN-gamma mRNA expression by MNCs. As previously reported for bovine polymorphonuclear neutrophils (PMNs), MNCs increased their expression of LFA-1, and their LKT binding and cytotoxicity, following exposure to IL-1beta, TNF-alpha, and IFN-gamma. These findings suggest that BHV-1 infection, and the resulting release of inflammatory cytokines, can stimulate expression of LFA-1 in bovine MNCs, thus enhancing the binding and biological effects of LKT. If such a mechanism occurs in vivo it might explain, in part, the increased susceptibility of BHV-1 infected cattle to bovine pasteurellosis. PMID- 15135986 TI - Apoptosis in normal lymphoid organs from healthy normal, conventional pigs at different ages detected by TUNEL and cleaved caspase-3 immunohistochemistry in paraffin-embedded tissues. AB - The frequency and the distribution of apoptotic cells were investigated in formalin-fixed paraffin-embedded lymphoid tissues from healthy conventional pigs at four different ages (6 days, 2 months, 3.5 months and 5 months). Samples of tonsil, mesenteric lymph node, spleen, thymus and Peyer's patches were histologically processed and apoptosis evaluated with the TUNEL reaction and cleaved caspase-3 immunohistochemistry. In each technique, quantification of positive labelling was done for each particular lymphoid tissue area. The labelling pattern and distribution were similar for TUNEL and cleaved caspase-3. TUNEL stained mainly apoptotic bodies inside macrophages, but signal was also seen in free apoptotic bodies and in the nuclei of lymphocyte-like cells. The anti-cleaved caspase-3 antibody labelled mainly nuclei of lymphocyte-like cells. All tissues presented a similar distribution pattern of apoptosis, except for the 6-day-old group. In this group, a scattered distribution of positive cells was detected in tonsil, lymph node and spleen. In the tonsil and mesenteric lymph nodes from the older pigs, follicular areas presented higher amounts of positive cells than interfollicular areas. Moreover, the splenic white pulp showed more positive reaction than the red pulp, especially when they included germinal centres. In all groups, the follicular areas of ileal Peyer's patches presented more labelled cells than the dome and the lamina propria. In the thymus, the higher apoptotic rates were found in the cortex. In general, TUNEL yielded higher rates of positive cells than cleaved caspase-3 immunolabelling. A good correlation between the two techniques was found for thymus, tonsil and mesenteric lymph node, but not for Peyer's patches and spleen. This study describes a detailed histochemical characterization of apoptosis in pig lymphoid tissues using TUNEL and a cleaved caspase-3 immunolabelling at different ages. Moreover, our results indicate that TUNEL and cleaved caspase-3 techniques can be equivalent only when tissues have a high or low levels of apoptosis, since a considerable discrepancy was found in intermediate situations. Data from this study should be useful for future comparative studies under disease conditions. PMID- 15135987 TI - BF, HP, DQB and DRB are associated with haemolytic complement activity, acute phase protein reaction and antibody response in the pig. AB - In order to examine the loci factor B (BF), C3, corticotropin releasing hormone (CRH), DQB, DRB, haptoglobin (HP) and transforming growth factor beta 1 (TGFB1) for association with traits of humoral, specific and unspecific defence F2 animals of a porcine resource family were genotyped at single nucleotide polymorphisms within these loci. Haemolytic complement activity in the alternative and classical pathway, C3c and haptoglobin serum concentration and antibody titres were determined immediately prior and at days 4 and 10 after vaccinations against Mycoplasma hyopneumoniae (Mh), Aujeszky's disease virus, and porcine reproductive and respiratory syndrome virus at 6, 14 and 16 weeks of age, respectively. Analysis of variance revealed association of BF, HP and DRB with C3c serum concentration. The trend of haemolytic complement activity and C3c serum concentration during the experiment was affected by the interaction of DQB genotype and time of measurement. Association with antibody titres were found for BF, DQB and DRB. Results of the mixed model analyses were confirmed by quantitative transmission disequilibrium test that showed linkage and association with antibody titres, complement activity and acute phase reaction at certain times of measurement. The findings promote the importance of the candidate genes for humoral mechanisms of unspecific and specific defence that provide natural resistance against many pathogens. PMID- 15135988 TI - Decreased sialylation of the acute phase protein alpha1-acid glycoprotein in feline infectious peritonitis (FIP). AB - Feline infectious peritonitis (FIP) is an immune-mediated disease of domestic and exotic felides infected with feline coronavirus. FIP is characterized by the overexpression of an acute phase protein, the alpha1-acid glycoprotein (AGP). In humans, AGP is a heavily glycosylated protein that undergoes several modifications of its glycan moiety during acute and chronic inflammatory pathologies. We studied the changes in AGP glycosylation in the course of FIP. Specifically, we focussed our attention on the degree of sialylation, fucosylation and branching. This study presents a purification method for feline AGP (fAGP) from serum, using an ion exchange chromatography strategy. The glycosylation pattern was analyzed in detail by means of interaction of purified fAGP with specific lectins. In particular, Sambucus nigra agglutinin I and Maackia amurensis agglutinin lectins were used to detect sialic acid residues, Aleuria aurantia lectin was used to detect L-fucose residues and Concanavalin A was used to evaluate the branching degree. By this method we showed that fAGP did not present any L-fucose residues on its surface, and that its branching degree was very low, both in normal and in pathological conditions. In contrast, during FIP disease, fAGP underwent several modifications in the sialic acid content, including decreased expression of both alpha(2-6)-linked and alpha(2-3)-linked sialic acid (76 and 44%, respectively when compared to non-pathological feline AGP). PMID- 15135989 TI - Modulation of the cytokine responses in equine macrophages following TACE inhibition. AB - The detrimental effects of the pro-inflammatory cytokine TNF-alpha during equine acute abdominal disease are well known. Its pivotal role in many human diseases has led to various in-depth studies regarding its release mechanism, in particular by TNF-alpha converting enzyme (TACE). In this study we investigated the inhibitory effect of a TACE-inhibitor on cytokine release (TNF-alpha, IL 1alpha and IL-6) in three different cell models, including U937 cells, a recently established equine macrophage cell line, known as eCAS, and primary equine PBMC. The aim was to show the similarity of TNF-alpha release through the TACE mechanism in human and equine cells after stimulation with LPS. Results indicate that, by using a TACE-inhibitor, TNF-alpha, IL-1alpha and IL-6 release can be reduced in both equine cell models and achieved comparable results in the human U937 cells. These results suggest that equine TNF-alpha, like its human homologue, is released from its membrane-bound position by TACE. PMID- 15135991 TI - A visual motion sensor based on the properties of V1 and MT neurons. AB - The motion response properties of neurons increase in complexity as one moves from primary visual cortex (V1), up to higher cortical areas such as the middle temporal (MT) and the medial superior temporal area (MST). Many of the features of V1 neurons can now be replicated using computational models based on spatiotemporal filters. However until recently, relatively little was known about how the motion analysing properties of MT neurons could originate from the V1 neurons that provide their inputs. This has constrained the development of models of the MT-MST stages which have been linked to higher level motion processing tasks such as self-motion perception and depth estimation. I describe the construction of a motion sensor built up in stages from two spatiotemporal filters with properties based on V1 neurons. The resulting composite sensor is shown to have spatiotemporal frequency response profiles, speed and direction tuning responses that are comparable to MT neurons. The sensor is designed to work with digital images and can therefore be used as a realistic front-end to models of MT and MST neuron processing; it can be probed with the same two dimensional motion stimuli used to test the neurons and has the potential to act as a building block for more complex models of motion processing. PMID- 15135992 TI - Effects of the noise level on induced motion. AB - Motion in a part of the field induces motion in an adjoining region. In this study, it was investigated how the noise level affects induced motion of a counterphase flickering (target) grating due to adjacent drifting (inducer) gratings. It was shown that at low noise levels, motion contrast occurred, and at high noise levels, motion assimilation occurred. When the noise level was randomly set for each trial, the adaptive change with the noise level was also observed. The result suggests that the adaptive change occurs for a short period. It was also found that noise for the target as well as noise for the inducers contributes to the effect of noise on motion induction. It suggests that the overall noise level is crucial for the effect. The study provided evidence that motion integration changes from a spatially band-pass operation to a low-pass operation as the signal-to-noise ratio (SNR) decreases. PMID- 15135993 TI - The peculiar nature of simultaneous colour contrast in uniform surrounds. AB - We present evidence from asymmetric colour matching experiments which strongly suggests that uniform surrounds evoke induction effects of a very peculiar nature, not representative of colour induction effects in variegated surrounds. Given the widespread use of uniform surrounds in studies of colour vision, this finding is of interest in relation to a number of current research issues, such as contrast coding of colour, functionally equivalent surrounds and colour constancy. A framework that systematises the seemingly complex colour appearance changes induced by uniform surrounds is presented and its implications are discussed. PMID- 15135994 TI - Investigating local network interactions underlying first- and second-order processing. AB - We compared the spatial lateral interactions for first-order cues to those for second-order cues, and investigated spatial interactions between these two types of cues. We measured the apparent modulation depth of a target Gabor at fixation, in the presence and the absence of horizontally flanking Gabors. The Gabors' gratings were either added to (first-order) or multiplied with (second-order) binary 2-D noise. Apparent "contrast" or modulation depth (i.e., the perceived difference between the high and low luminance regions for the first-order stimulus, or between the high and low contrast regions for the second-order stimulus) was measured with a modulation depth-matching paradigm. For each observer, the first- and second-order Gabors were equated for apparent modulation depth without the flankers. Our results indicate that at the smallest inter element spacing, the perceived reduction in modulation depth is significantly smaller for the second-order than for the first-order stimuli. Further, lateral interactions operate over shorter distances and the spatial frequency and orientation tuning of the suppression effect are broader for second- than first order stimuli. Finally, first- and second-order information interact in an asymmetrical fashion; second-order flankers do not reduce the apparent modulation depth of the first-order target, whilst first-order flankers reduce the apparent modulation depth of the second-order target. PMID- 15135995 TI - Contour interpolation revealed by a dot localization paradigm. AB - Contour interpolation mechanisms allow perception of bounded objects despite incomplete edge information. Here, we introduce a paradigm that maps interpolated contours as they unfold over time. Observers localize dots relative to perceived boundaries of illusory, partly occluded, or control stimuli. Variations in performance with dot position and processing time reveal the location and precision of emerging contour representations. Illusory and occluded contours yielded more proficient dot localization than control stimuli containing only spatial cues, suggesting performance based on low-level representations. Further, illusory contours exhibited a distinct developmental time course, emerging over the first 120 ms of processing. These experiments establish the effectiveness of the dot localization paradigm for examining interpolated edge representations, contour microgenesis, and the underlying processing mechanisms. PMID- 15135996 TI - Learning motion discrimination with suppressed MT. AB - We studied perceptual learning in motion discrimination when the brain's middle temporal area (MT/V5) was functionally suppressed. This was achieved by using the "paired-dots" motion stimulus where the two dots in a pair always move in counter phase over a short distance [J. Neurosci. 14 (1994) 7357]. The motion directional signal of the stimulus is therefore always 0 on average. As a result, this stimulus activates MT in Rhesus monkeys no more than flicker noise does [J. Neurosci. 14 (1994) 7367]. We added a new manipulation to eliminate the Glass pattern in the original stimulus that would have otherwise provided a static orientation cue. Two such new motion stimuli were presented sequentially, in a 2AFC task. Subjects decided if the global motion-axis of the stimuli changed clockwise or counter-clockwise. When the task difficulty was set at 60% correct, none of the subjects could learn with feedback, even though their performance was well above chance. However, when the task difficulty was set instead at 70% correct, a new group of subjects was able to learn. Hence, learning motion discrimination was possible when MT was presumably eliminated. PMID- 15135997 TI - Lightness constancy through transparency: internal consistency in layered surface representations. AB - Asymmetric lightness matching was employed to measure how the visual system assigns lightness to surface patches seen through partially-transmissive surfaces. Observers adjusted the luminance of a comparison patch seen through transparency, in order to match the lightness of a standard patch seen in plain view. Plots of matched-to-standard luminance were linear, and their slopes were consistent with Metelli's alpha. A control experiment confirmed that these matches were indeed transparency based. Consistent with recent results, however, when observers directly matched the transmittance of transparent surfaces, their matches deviated strongly and systematically from Metelli's alpha. Although the two sets of results appear to be contradictory, formal analysis reveals a deeper mutual consistency in the representation of the two layers. A ratio-of-contrasts model is shown to explain both the success of Metelli's model in predicting lightness through transparency, and its failure to predict perceived transmittance--and hence is seen to play the primary role in perceptual transparency. PMID- 15135998 TI - Influence of cone pigment coexpression on spectral sensitivity and color vision in the mouse. AB - The mouse retina contains both middle-wavelength-sensitive (M) and ultraviolet sensitive (UV) photopigments that are coexpressed in cones. To examine some potential visual consequences of cone pigment coexpression, spectral sensitivity functions were measured in mice (Mus musculus) using both the flicker electroretinogram (ERG) and behavioral discrimination tests. Discrimination tests were also employed to search for the presence of color vision in the mouse. Spectral sensitivity functions for the mouse obtained from ERG measurements and from psychophysical tests each reveal contributions from two classes of cone having peak sensitivities (lambda(max)) of approximately 360 and 509-512 nm. The relative contributions of the two pigment types to spectral sensitivity differ significantly in the two types of measurements with a relationship reversed from that often seen in mammals. Mice were capable of discriminating between some pairs of spectral stimuli under test conditions where luminance-related cues were irrelevant. Since mice can make dichromatic color discriminations, their visual systems must be able to exploit differences in the spectral absorption properties among the cones. Complete selective segregation of opsins into individual photoreceptors is apparently not a prerequisite for color vision. PMID- 15135999 TI - Treatment of astigmatism-related amblyopia in 3- to 5-year-old children. AB - Best-corrected acuity was measured for vertical and horizontal gratings and for Lea Symbols recognition acuity in 3- to 5-year-old children with high astigmatism and in non-astigmatic children. There was significant amblyopia among astigmatic children at baseline. There was no evidence that eyeglass correction of astigmatism resulted in a reduction in amblyopia over a 4-month average treatment duration (although vision in astigmatic children was significantly improved immediately upon eyeglass correction, indicating that eyeglass correction did provide a visual benefit). Treatment outcome results are discussed in terms of both methodological issues and theoretical implications. PMID- 15136000 TI - Center-surround interactions in visual motion processing during binocular rivalry. AB - When each eye is confronted with a dissimilar stimulus, the percept will generally alternate between the two. This phenomenon is known as binocular rivalry. Although binocular rivalry occurs at locations where targets overlap spatially, the area surrounding rivalrous targets can modulate their dominance. Here we show that during binocular rivalry of oppositely moving gratings, a surrounding grating moving in the same direction as one of the two leads to increased dominance of the opposite direction of motion in the center. This increased dominance of the opposite direction in the center was observed irrespective of the eye to which the surround was presented. Inspection of the results for different conditions reveals that the preference for the opposite direction of motion cannot be explained by a single mechanism operating beyond binocular fusion. We therefore suggest that this phenomenon is the outcome of center-surround interactions at multiple levels along the pathway of visual motion processing. PMID- 15136001 TI - Comparison of different models of orientation selectivity based on distinct intracortical inhibition rules. AB - Aim of this work is to present simple models of orientation selectivity in the visual cortex, which do not require massive computational effort. Three different models are compared, in order to gain deeper insight into the structure of cortical circuits generating inhibitory signals. All models represent a single hypercolumn. They differ as to the arrangement of inhibitory connections: in the first ("antiphase inhibition model") inhibition is in phase opposition with excitation, but with a similar orientation tuning; in the second ("in-phase inhibition model"), inhibition is in phase with excitation, but with larger orientation tuning. In these two models the orientation width of inhibition increases with contrast. Finally, a third model ("center-surround model") assumes that inhibition comes from the same cells providing excitation, hence the inhibition tuning is contrast-independent. All models, with suitable values of the intracortical synapse parameters, are able to mimic experimental results in the literature. A few differences are evident between the "center-surround model" and the other two, especially as to the dependence of cortical cell response on spatial frequency. The models can represent practical tools to test hypotheses on the disposition of cortical synapses avoiding massive computational efforts. PMID- 15136002 TI - Electrophysiological correlates of lateral interactions in human visual cortex. AB - Detection thresholds for visually presented targets can be influenced by the nature of information in adjacent regions of the visual field. For example, detection thresholds for low-contrast Gabor patches decrease when flanked by patches that are oriented collinearly rather than orthogonally with the target. Such results are consistent with the known microanatomy of primary visual cortex, where long-range horizontal connections link cortical columns with common orientation preferences. To investigate the neural bases of collinearity effects, we recorded event-related brain potentials (ERPs) together with psychophysical measures for targets flanked by collinear vs. orthogonal gratings. Human volunteers performed a contrast discrimination task on a target grating presented at a perifoveal location. For targets flanked by collinear stimuli, we observed an increased positive polarity voltage deflection in the occipital scalp-recorded ERPs between 80 to 140 ms after stimulus onset. Such a midline occipital scalp voltage distribution of this ERP collinearity effect is consistent with a generator in primary visual cortex. Two later negative voltage ERP deflections (latencies of 245-295 and 300-350 ms) were focused at lateral occipital scalp sites, a pattern consistent with activity in extrastriate visual cortex. These ERP effects were correlated with improved contrast discrimination for central targets presented with collinear flanks. These results demonstrate that the integration of local flanking elements with a central stimulus can occur as early as 80 ms in human visual cortex, but this includes processes occurring at longer latencies and appears to involve both striate and extrastriate visual areas. PMID- 15136003 TI - Grouping and segmentation in binocular rivalry. AB - Dichoptic presentation of dot arrays produces binocular rivalry if the arrays are of opposite contrast relative to background. Rivalry can occur even if individual dots in one eye's array do not overlap with the dots in the contralateral eye's array. The amount of unitary perception of only one array is a measure of the probability that the stimuli rival as textured surfaces rather than as portions of arrays or as individual dot elements. In accordance with Gestalt grouping principles, arrays of uniform brightness or color produced more unitary perception than mixed arrays. However, experiments with parametric variation of dot motion coherence suggested that segmentation mechanisms based on detection of collinearity can also influence perceptual selection and suppression in binocular rivalry. PMID- 15136004 TI - Simulation of artificial vision: II. Eccentric reading of full-page text and the learning of this task. AB - Reading of isolated words in conditions mimicking artificial vision has been found to be a difficult but feasible task. In particular at relatively high eccentricities, a significant adaptation process was required to reach optimal performances [Vision Res. 43 (2003) 269]. The present study addressed the task of full-page reading, including page navigation under control of subject's own eye movements. Conditions of artificial vision mimicking a retinal implant were simulated by projecting stimuli with reduced information content (lines of pixelised text) onto a restricted and eccentric area of the retina. Three subjects, naive to the task, were trained for almost two months (about 1 h/day) to read full-page texts. Subjects had to use their own eye movements to displace a 10 degrees x 7 degrees viewing window, stabilised at 15 degrees eccentricity in their lower visual field. Initial reading scores were very low for two subjects (about 13% correctly read words), and astonishingly high for the third subject (86% correctly read words). However, all of them significantly improved their performance with time, reaching close to perfect reading scores (ranging from 86% to 98% correct) at the end of the training process. Reading rates were as low as 1-5 words/min at the beginning of the experiment and increased significantly with time to 14-28 words/min. Qualitative text understanding was also estimated. We observed that reading scores of at least 85% correct were necessary to achieve 'good' text understanding. Gaze position recordings, made during the experimental sessions, demonstrated that the control of eye movements, especially the suppression of reflexive vertical saccades, constituted an important part of the overall adaptive learning process. Taken together, these results suggest that retinal implants might restore full-page text reading abilities to blind patients. About 600 stimulation contacts, distributed on an implant surface of 3 x 2 mm2, appear to be a minimum to allow for useful reading performance. A significant learning process will however be required to reach optimal performance with such devices, especially if they have to be placed outside the foveal area. PMID- 15136005 TI - Capacity limitations of visual memory in two-interval comparison of Gabor arrays. AB - The capacity of short-term visual memory (VSTM) was assessed in a two-interval spatial frequency (SF) discrimination task. The cued Gabor target in a multi element array either increased or decreased in SF across a 2 s inter-stimulus interval (ISI). Distracters as well as target were made to change across ISI so that memory of the individual SF of Gabor elements was required to solve the discrimination. The dynamics of the information loss from visual memory were analysed by manipulating the timing of spatial cues and masks. Cueing the target position before the first display gave thresholds comparable with those for a single Gabor patch. Cues placed after the first display gave higher thresholds indicating some loss of information. Within the ISI there was little increase in threshold or set size effect with cue delay. However there was a sharp rise in thresholds for cue positions after the second display. Gabor masks placed before a mid-ISI cue were more effective than noise masks or Gabor masks placed after the cue. With a cue placed late in the ISI, preceded by a Gabor mask, the masking effect decreased with increasing delay of the mask after the first display. This suggests a selective, dynamic but increasingly durable representation of the initial stimulus is built up in memory, and there is a graded form of "overwriting" of this representation by new stimuli. PMID- 15136006 TI - Rotation direction affects object recognition. AB - What role does dynamic information play in object recognition? To address this question, we probed observers' memory for novel objects rotating in depth. Irrespective of object discriminability, performance was affected by an object's rotation direction. This effect was obtained despite the same shape information and views being shown for different rotation directions. This direction effect was eliminated when either static images or animations that did not depict globally coherent rotation were used. Overall, these results suggest that dynamic information, that is, the spatiotemporal ordering of object views, provides information independent of shape or view information to a recognition system. PMID- 15136007 TI - Demography in the 21st century: introduction. PMID- 15136008 TI - Markov mortality models: implications of quasistationarity and varying initial distributions. AB - This paper explains some implications of Markov-process theory for models of mortality. We show that an important qualitative feature common to empirical mortality data, and which has been found in certain models-the convergence to a "mortality plateau"-is, in fact, a generic consequence of the models' convergence to a "quasistationary distribution". This phenomenon has been explored extensively in the mathematical literature. Not only does this generalization free important results from specifics of the models, it also suggests a new explanation for the convergence to constant mortality. At the same time that we show that the late behavior of these models-convergence to a finite asymptote-is almost logically immutable, we also point out that the early behavior of the mortality rates can be more flexible than has been generally acknowledged. We show, in particular, that an appropriate choice of initial conditions enables one popular model to approximate any reasonable hazard-rate data. This illustrates how precarious it can be to read a model's vindication from its consilience with a favored hazard-rate function, such as the Gompertz exponential. PMID- 15136009 TI - The case for negative senescence. AB - Negative senescence is characterized by a decline in mortality with age after reproductive maturity, generally accompanied by an increase in fecundity. Hamilton (1966) ruled out negative senescence: we adumbrate the deficiencies of his model. We review empirical studies of various plants and some kinds of animals that may experience negative senescence and conclude that negative senescence may be widespread, especially in indeterminate-growth species for which size and fertility increase with age. We develop optimization models of life-history strategies that demonstrate that negative senescence is theoretically possible. More generally, our models contribute to understanding of the evolutionary and demographic forces that mold the age-trajectories of mortality, fertility and growth. PMID- 15136010 TI - The shape of things to come: using models with physiological structure to predict mortality trajectories. AB - If mortality rate is viewed as the outcome of processes of behavior, growth and reproduction, then it should be possible to predict mortality rate as a result of those processes. We provide two examples of how this may be done. In the first, we use the method of linear chains to treat mortality that is the result of multiple physiological processes, some of which may have delays. In the second, we assume that mortality is the result of damage associated with growth and metabolism. Both approaches lead to a rich diversity of predicted mortality trajectories. Although many of these look Gompertzian at young ages, the behavior at older ages depends upon the details of the physiological models. PMID- 15136011 TI - The effects of background mortality on optimal reproduction in a seasonal environment. AB - We consider optimal annual routines of reproductive behaviour in a seasonal environment. In our model the condition of the organism is adversely affected by hard work, but can recover during easy periods. Our analysis concentrates on the effects of background mortality (i.e., mortality that cannot be avoided) on the optimal strategy and how often an organism following this strategy breeds. In particular, we are concerned with whether reproduction occurs at specific times of year (entrained to the annual cycle), and if so then how many reproductive bouts occur per year. We find that an increase in background mortality can have various effects. If the animal is entrained to the annual cycle and has one breeding attempt per year, then breeding tends to occur earlier and there may be two breeding attempts per season. Another possible outcome is that breeding is no longer entrained. If the animal is entrained but sometimes skips reproduction so that it does not breed every year, then an increase in mortality may make it more likely that the animal breeds every year. We show that as background mortality increases the resultant increase in the frequency of breeding contributes to the increase in annual mortality. We also explore the effects of mortality on the timing of reproduction within a year, highlighting the tension between the interests of the parent and that of the young. PMID- 15136012 TI - Age-structured optimal control in population economics. AB - This paper brings both intertemporal and age-dependent features to a theory of population policy at the macro-level. A Lotka-type renewal model of population dynamics is combined with a Solow/Ramsey economy. We consider a social planner who maximizes an aggregate intertemporal utility function which depends on per capita consumption. As control policies we consider migration and saving rate (both age-dependent). By using a new maximum principle for age-structured control systems we derive meaningful results for the optimal migration and saving rate in an aging population. The model used in the numerical calculations is calibrated for Austria. PMID- 15136013 TI - Estimating national disability risk. AB - In this article, I provide a method to rebuild the active and disabled life expectancy (ALE and DLE) on the basis of 'current' death and disability risks, and to measure disability risk. This method uses national-level data, and is based on two main assumptions. The first is the Gompertz assumption that death rate rises with age exponentially, and the second is the Cox assumption that death rates of active status are proportional to those of disabled status across age. Applying this method to the US data, I find that the disability risk has increased between 1970 and 1990 for both men and women aged 40 and older. Situations in which above assumptions could be removed are also discussed. PMID- 15136014 TI - Elasticity analysis of density-dependent matrix population models: the invasion exponent and its substitutes. AB - In density-independent models, the population growth rate lambda measures population performance, and the perturbation analysis of lambda (its sensitivity and elasticity) plays an important role in demography. In density-dependent models, the invasion exponent lambdaI replaces lambda as a measure of population performance. The perturbation analysis of lambdaI reveals the effects of environmental changes and management actions, gives the direction and intensity of density-dependent natural selection on life history traits, and permits calculation of the sampling variance of the invasion exponent. Because density dependent models require more data than density-independent models, it is tempting to look for substitutes for the invasion exponent, the sensitivity and elasticity of which can be calculated from a density-independent model. Here we examine the accuracy of two such substitutes: the dominant eigenvalue of the projection matrix evaluated at equilibrium (An) and the dominant eigenvalue of the matrix averaged over the attractor (A). Using a two-stage model that represents a wide range of life history types, we find that the elasticities of An or A often agree to within less than 5% error with those of the invasion exponent, even when population dynamics are chaotic. The exceptions are for semelparous life histories, especially when density-dependence affects fertility. This suggests that the elasticity analysis of density-independent models near equilibrium, or averaged over the attractor, provides useful information about the elasticity of the invasion exponent in density-dependent models. PMID- 15136015 TI - Likelihood-based inference for stochastic models of sexual network formation. AB - Sexually-transmitted diseases (STDs) constitute a major public health concern. Mathematical models for the transmission dynamics of STDs indicate that heterogeneity in sexual activity level allow them to persist even when the typical behavior of the population would not support endemicity. This insight focuses attention on the distribution of sexual activity level in a population. In this paper, we develop several stochastic process models for the formation of sexual partnership networks. Using likelihood-based model selection procedures, we assess the fit of the different models to three large distributions of sexual partner counts: (1) Rakai, Uganda, (2) Sweden, and (3) the USA. Five of the six single-sex networks were fit best by the negative binomial model. The American women's network was best fit by a power-law model, the Yule. For most networks, several competing models fit approximately equally well. These results suggest three conclusions: (1) no single unitary process clearly underlies the formation of these sexual networks, (2) behavioral heterogeneity plays an essential role in network structure, (3) substantial model uncertainty exists for sexual network degree distributions. Behavioral research focused on the mechanisms of partnership formation will play an essential role in specifying the best model for empirical degree distributions. We discuss the limitations of inferences from such data, and the utility of degree-based epidemiological models more generally. PMID- 15136016 TI - Hepatobiliary imaging and its pitfalls. AB - In summary, US is the initial imaging modality for the evaluation of acute right upper quadrant pain. It permits accurate diagnosis of acute cholecystitis and successfully identifies multiple other causes of patient symptomatology. Some of these processes lie outside the hepatobiliary system and include renal infection and obstruction, pancreatitis and its sequelae, duodenal or colonic perforation or mass lesions, peritoneal tumor spread, adrenal hemorrhage, and even remote problems, such as pneumonia. The limitations on US include incomplete imaging of the liver, most often at the dome or beneath ribs on the surface, and incomplete visualization of lesion boundaries, particularly with some infections and tumors. For these clinical scenarios, contrast-enhanced CT is complementary to US and should be encouraged. In the biliary tree, US has limitations in situations in which the ducts are not dilated and sometimes with imaging the extra hepaticducts, especially distally. For these patients, CT or MR imaging (MRCP) is especially useful. If one keeps the clinical scenario in mind and always images a patient where he or she hurts, US is a powerful and effective diagnostic method for evaluating acute right upper quadrant pain. PMID- 15136017 TI - Venous protocols, techniques, and interpretations of the upper and lower extremities. AB - Deep venous thrombosis and pulmonary embolism are common problems and frequently included in the differential diagnosis of patients presenting to the emergency department and in the acute care setting. Clinical signs and symptoms of these entities are nonspecific and it is important to perform objective testing to confirm the diagnosis and initiate proper therapy. Compression ultrasonography is the diagnostic procedure of choice for the investigation of patients with suspected deep venous thrombosis. This article reviews the clinical indications, diagnostic techniques, and interpretation of compression ultrasound to assess deep venous thrombosis in the upper and lower extremities and to evaluate the role of compression ultrasonography in the assessment of patients with suspected pulmonary embolism. PMID- 15136018 TI - Sonographic evaluation of first-trimester bleeding. AB - Vaginal bleeding is a leading cause of presentation for emergency care during the first trimester of the pregnancy. Clinical assessment of the pregnancy outcome at this stage is less reliable. US examination is crucial in establishing IUP and early pregnancy failure and to exclude other causes of bleeding, such as ectopic pregnancy and molar pregnancy. Diagnosis of a normal IUP at this stage not only assists the physician in an expectant management, but also gives a psychologic boost to the patient. With recent advances in US technology and the availability of high-frequency transvaginal transducers, reliable diagnosis of early pregnancy failure can be made even before the embryo is visible. PMID- 15136019 TI - The role of ultrasound in pregnancy-related emergencies. AB - Most complications of pregnancy allow time for transfer to specialized obstetric ultrasound units, but many women present to the emergency room or the labor and delivery unit with signs and symptoms suggesting genuine acute medical emergencies, where successful outcome depends on prompt diagnosis of the disorder and rapid appropriate medical management. The use of ultrasound technology in obstetric emergencies is well established. Ultrasonography plays a major role in such cases as the most important tool clinicians are using to identify the correct etiology and diagnosis, whereas in other cases it helps limit the differential diagnosis. One of the goals of any advanced training program in obstetrics and gynecology and radiology is to allow the skilled physician to perform the proper ultrasound study in case of an obstetric emergency to facilitate the proper diagnosis, enabling the medical team to provide the best possible care. PMID- 15136020 TI - Adnexal mass with pelvic pain. AB - In a pregnant woman who presents with acute pelvic pain and an adnexal mass, pregnancy-related etiologies, such as ectopic pregnancy or ovarian torsion, are typically the first diagnoses to be considered. Many other causes of pelvic pain associated with an adnexal mass can occur in pregnant patients, however. Some causes are benign and others require urgent management and treatment. Clinical presentation and physical examination can be misleading in pregnancy. The location of pain may be atypical for the pathologic entity, the pain may be muted, and in the case of infection, fever and leukocytosis can be absent. US examination is a safe and effective method for evaluating these patients. Sonographic characterization of adnexal masses may make a definitive diagnosis or focus the differential, which allows for prompt and appropriate treatment of patients. PMID- 15136021 TI - Acute painful scrotum. AB - The ability of US to diagnose the pathogenesis of the acute scrotum is unsurpassed by any other imaging modality. It is the first imaging performed in patients with acute scrotum. Knowledge of the normal and pathologic sonographic appearance of the scrotum and proper sonographic technique is essential for accurate diagnosis of acute scrotum. High-frequency transducer sonography combined with color flow Doppler sonography provides the information essential to reach a specific diagnosis in patients with testicular torsion, epididymo orchitis, and testicular trauma. PMID- 15136022 TI - Ultrasound evaluation of abdominal aortic and iliac aneurysms and mesenteric ischemia. AB - The role of US in imaging of the abdominal vasculature has broadened over recent years. Long considered the modality of choice in the detection of AAA, its use has expanded to diagnosing and monitoring IAAs and PAAs, screening for mesenteric ischemia, and posttreatment monitoring of endovascular stents. PMID- 15136023 TI - Arterial injuries: a sonographic approach. AB - Acute arterial emergencies can arise from direct traumatic injury to the artery or be spontaneous. This article emphasizes the various presentations of arterial emergencies. These include acute arterial occlusions; excessive bleeding; and hematoma formation caused by penetrating arterial wall injuries, pseudoaneurysms, and arteriovenous fistulas. The broad category of arterial occlusions includes traumatic lacerations, embolizations, and arterial dissections. Modern ultrasound equipment is a rapid and convenient imaging approach in many of these clinical scenarios. In combination with MR angiography and CT angiography, these noninvasive tests can diagnose the presence of most arterial injuries, and be used to measure their impact. PMID- 15136024 TI - Emergency Doppler evaluation of the liver and kidneys. AB - Doppler ultrasound is useful in the emergent evaluation of the liver and kidney transplant patient. Arterial stenosis, pseudoaneurysm, and venous thrombosis are treatable causes of allograft failure that can be detected easily with color and spectral Doppler. Doppler has a limited but important role in the emergent evaluation of the native liver and kidneys, usually involving prior biopsy or instrumentation. PMID- 15136025 TI - Emergency ultrasound in trauma patients. AB - US will be used more frequently in the future for the evaluation of traumatized patients. Previously,the main focus of the sonographic examination was for the detection of free fluid. Unstable patients with free fluid often can be triaged to the operation room without further imaging tests. In patients who are more stable or in whom US results are negative, CT is required. Based on recent studies, sonography has a sensitivity rate of approximately 40% in direct detection of solid organ injuries. In the future, however, with the use of contrast-enhanced agents, sonography may more reliably detect solid organ injuries. Within the chest, US has been shown to be helpful in detecting pleural effusions and may be useful in detecting pericardial effusions. US has been shown to be sensitive in detecting pneumothoraces in traumatized patients. PMID- 15136026 TI - Priapism. AB - Priapism is a relatively uncommon condition that may present as a medical emergency associated with significant pain and anxiety in the veno-occlusive or low-flow variant. Pharmacologic advances and, specifically, the availability of intracavemosal alpha-agonist therapy have dramatically improved the prospects of resolution for patients with low-flow priapism presenting within the first few hours of the acute episode. High-flow priapism is not considered an emergency and treatment measures are typically conservative aimed at preservation of potency. Urologists, radiologists, and other health care personnel caring for the patient with priapism must be familiar with various etiologic factors implicated in low flow and high-flow priapism to formulate a logical step-care approach. Differentiation of the low-flow from the high-flow state is perhaps the most critical initial diagnostic challenge that determines the sequence of further interventions including surgical shunts in low-flow priapism refractory to medical therapy. PMID- 15136027 TI - Ultrasound evaluation of acute abdominal emergencies in infants and children. AB - Ultrasound is extremely beneficial in the evaluation of acute pediatric abdominal disease, such as HPS, intussusception, and acute appendicitis. As techniques and equipment improve, its role in the evaluation of infants and children continues to increase. PMID- 15136028 TI - Emergent ultrasound interventions. AB - The interventionist can perform many emergent procedures with ultrasound guidance, because of its real-time, multiplanar imaging capability and portability. With the use of color Doppler, additional important information, such as aberrant vessels, can be ascertained to help plan needle trajectory. Ultrasound is also useful for nonemergent procedures, such as biopsies. All interventionists are encouraged to be facile with the use of ultrasound. PMID- 15136030 TI - X-ray crystal structure of a non-crystalline cellulose-specific carbohydrate binding module: CBM28. AB - Natural cellulose exists as a composite of different forms, which have historically been broadly characterized as "crystalline" or "amorphous". The recognition of both of these forms of cellulose by the carbohydrate-binding modules (CBM) of microbial glycoside hydrolases is central to natural and efficient biotechnological conversion of plant cell wall biomass. There is increasing evidence that, at least some, individual binding modules target distinct and different regions of non-crystalline "amorphous" cellulose. Competition experiments show that CBM28 modules do not compete with CBM17 modules when binding to non-crystalline cellulose. The structure of the BspCBM28 (http://afmb.cnrs-mrs.fr/CAZY/) module from the Bacillus sp. 1139 family GH5 endoglucanase, comprising a 191 amino acid protein, has therefore been determined at 1.4A resolution using single isomorphous replacement with anomalous scattering methods. The structure reveals a "beta-jelly roll" topology, with high degree of similarity to the structure of CBM17 domains. Sequence and structural conservation strongly suggests that these two families of domains have evolved through gene duplication and subsequent divergence. The ligand-binding site "topographies" of CBMs from families 28, 17 and 4 begins to shed light on the differential recognition of non-crystalline cellulose by multi-modular plant cell wall-degrading enzymes. PMID- 15136031 TI - The N-terminal prion domain of Ure2p converts from an unfolded to a thermally resistant conformation upon filament formation. AB - According to the "amyloid backbone" model of Ure2p prionogenesis, the N-terminal domain of Ure2p polymerizes to form an amyloid filament backbone surrounded by the C-terminal domains. The latter domains retain their native glutathione-S transferase (GST)-like fold but are sterically inactivated from their regulatory role in nitrogen catabolism. We have tested this model by differential scanning calorimetry of soluble and filamentous Ure2p and of soluble C-terminal domains, combined with electron microscopy. As predicted, the C-terminal domains respond to thermal perturbation identically in all three states, exhibiting a single endotherm at 76 degrees C. In contrast, no thermal signal was associated with the N-terminal domains: in the soluble state of Ure2p, because they are unfolded; in the filamentous state, because their robust amyloid conformation resists heating to 100 degrees C. PMID- 15136032 TI - Analysis of the open and closed conformations of the GTP-binding protein YsxC from Bacillus subtilis. AB - Genetic analysis has suggested that the product of the Bacillus subtilis ysxC gene is essential for survival of the microorganism and hence may represent a target for the development of a novel anti-infective agent. B.subtilis YsxC is a member of the translation factor related class of GTPases and its crystal structure has been determined in an apo form and in complex with GDP and GMPPNP/Mg2+. Analysis of these structures has allowed us to examine the conformational changes that occur during the process of nucleotide binding and GTP hydrolysis. These structural changes particularly affect parts of the switch I and switch II region of YsxC, which become ordered and disordered, respectively in the "closed" or "on" GTP-bound state and disordered and ordered, respectively, in the "open" or "off" GDP-bound conformation. Finally, the binding of the magnesium cation results in subtle shifts of residues in the G3 region, at the start of switch II, which serve to optimize the interaction with a key aspartic acid residue. The structural flexibility observed in YsxC is likely to contribute to the role of the protein, possibly allowing transduction of an essential intracellular signal, which may be mediated via interactions with a conserved patch of surface-exposed, basic residues that lies adjacent to the GTP-binding site. PMID- 15136033 TI - The role of prophage-like elements in the diversity of Salmonella enterica serovars. AB - The Salmonella enterica serovar Typhi CT18 (S.Typhi) chromosome harbours seven distinct prophage-like elements, some of which may encode functional bacteriophages. In silico analyses were used to investigate these regions in S.Typhi CT18, and ultimately compare these integrated bacteriophages against 40 other Salmonella isolates using DNA microarray technology. S.Typhi CT18 contains prophages that show similarity to the lambda, Mu, P2 and P4 bacteriophage families. When compared to other S.Typhi isolates, these elements were generally conserved, supporting a clonal origin of this serovar. However, distinct variation was detected within a broad range of Salmonella serovars; many of the prophage regions are predicted to be specific to S.Typhi. Some of the P2 family prophage analysed have the potential to carry non-essential "cargo" genes within the hyper-variable tail region, an observation that suggests that these bacteriophage may confer a level of specialisation on their host. Lysogenic bacteriophages therefore play a crucial role in the generation of genetic diversity within S.enterica. PMID- 15136034 TI - The 12 A structure of trypsin-treated measles virus N-RNA. AB - Recombinant measles virus nucleoprotein (N) was produced in insect cells where it bound to cellular RNA to form helical N-RNA structures. These structures were observed by electron microscopy but were too flexible for high-resolution image analysis. Removal of the C-terminal tail of N by trypsin treatment resulted in structures that were much more rigid and seemed more regular. Several methods of image analysis were employed in order to make a helical reconstruction of the digested N-RNA. During this analysis, it became clear that the apparently regular coils of digested N-RNA consisted of a series of closely related helical states. The iterative helical real space reconstruction method allowed the identification of two helical states for which a reconstruction could be calculated. The model with the highest resolution shows N monomers that consist of three domains and that are connected to their neighbours by two narrow connections, one close to the helical axis and another toward the middle of the monomers. There are no connections between N molecules in subsequent helical turns. After labelling the RNA in the structure with cis-platinum, the connection closest to the helical axis increased in density, suggesting the position of the RNA. The shapes of the monomers of the nucleoproteins of influenza virus, rabies virus (both determined before) and that of measles virus (determined here) are all similar, whereas the overall shapes of their respective N-RNAs (nucleocapsids) is very different. This is probably due to the position and number of the connections between the N subunits in the N-RNA, one for influenza virus allowing much flexibility, two for rabies virus at either end of the N molecules leading to ribbons and two for measles virus leading to the typical paramyxovirus helical nucleocapsid. PMID- 15136035 TI - The Ankrd2 protein, a link between the sarcomere and the nucleus in skeletal muscle. AB - Ankrd2 may be a link between the sarcomere and the nucleus; a similar role has recently been proposed for CARP that has a high level of structural and functional conservation with Ankrd2. Both Ankrd2 and CARP are involved in striated muscle hypertrophy. The mechanism by which muscle stretch is sensed and signals are transduced is still unknown; however, Ankrd2 and CARP could play similar roles in pathways leading to hypertrophy, the triggering mechanisms being heart pressure overload monitored by CARP and mechanical stretch in skeletal muscle monitored by Ankrd2. Recently Ankrd2 and CARP have been proposed as members of a family of muscle ankyrin repeat proteins (MARPs) that form a complex with titin, myopalladin and calpain protease p94, involved in signaling and regulation of gene expression in response to muscle stress. Here, we show that Ankrd2 is able to interact with the Z-disc protein telethonin as well as being able to interact with three transcription factors: YB-1, PML and p53. Ankrd2 binding to the ubiquitous transcription factor YB-1 can be demonstrated both in vitro and in vivo; this is not very surprising, since a similar interaction was previously described for CARP. However, the interactions with PML and p53 are unexpected new findings, with interesting implications in the Ankrd2 signaling cascade. Ankrd2 co-localizes with the transcriptional co-activator and co repressor PML in nuclear bodies (NBs) in human myoblasts as detected by confocal immunofluorescence. Interestingly, we show that Ankrd2 not only binds the tumor suppressor protein p53 both in vitro and in vivo but also enhances the up regulation of the p21(WAFI/CIPI) promoter by p53. Therefore, our findings strengthen the hypothesis that Ankrd2 may be involved in sensing stress signals and linking these to muscle gene regulation. PMID- 15136036 TI - Molecular evolution of a microRNA cluster. AB - Many of the known microRNAs are encoded in polycistronic transcripts. Here, we reconstruct the evolutionary history of the mir17 microRNA clusters which consist of miR-17, miR-18, miR-19a, miR-19b, miR-20, miR-25, miR-92, miR-93, miR-106a, and miR-106b. The history of this cluster is governed by an initial phase of local (tandem) duplications, a series of duplications of entire clusters and subsequent loss of individual microRNAs from the resulting paralogous clusters. The complex history of the mir17 microRNA family appears to be closely linked to the early evolution of the vertebrate lineage. PMID- 15136037 TI - Crystal structure of KsgA, a universally conserved rRNA adenine dimethyltransferase in Escherichia coli. AB - The bacterial enzyme KsgA catalyzes the transfer of a total of four methyl groups from S-adenosyl-l-methionine (S-AdoMet) to two adjacent adenosine bases in 16S rRNA. This enzyme and the resulting modified adenosine bases appear to be conserved in all species of eubacteria, eukaryotes, and archaebacteria, and in eukaryotic organelles. Bacterial resistance to the aminoglycoside antibiotic kasugamycin involves inactivation of KsgA and resulting loss of the dimethylations, with modest consequences to the overall fitness of the organism. In contrast, the yeast ortholog, Dim1, is essential. In yeast, and presumably in other eukaryotes, the enzyme performs a vital role in pre-rRNA processing in addition to its methylating activity. Another ortholog has been discovered recently, h-mtTFB in human mitochondria, which has a second function; this enzyme is a nuclear-encoded mitochondrial transcription factor. The KsgA enzymes are homologous to another family of RNA methyltransferases, the Erm enzymes, which methylate a single adenosine base in 23S rRNA and confer resistance to the MLS-B group of antibiotics. Despite their sequence similarity, the two enzyme families have strikingly different levels of regulation that remain to be elucidated. We have crystallized KsgA from Escherichia coli and solved its structure to a resolution of 2.1A. The structure bears a strong similarity to the crystal structure of ErmC' from Bacillus stearothermophilus and a lesser similarity to sc mtTFB, the Saccharomyces cerevisiae version of h-mtTFB. Comparison of the three crystal structures and further study of the KsgA protein will provide insight into this interesting group of enzymes. PMID- 15136038 TI - Gaining target access for deoxyribozymes. AB - Antisense oligonucleotides and ribozymes have been used widely to regulate gene expression by targeting mRNAs in a sequence-specific manner. Long RNAs, however, are highly structured molecules. Thus, up to 90% of putative cleavage sites have been shown to be inaccessible to classical RNA based ribozymes or DNAzymes. Here, we report the use of modified nucleotides to overcome barriers raised by internal structures of the target RNA. In our attempt to cleave a broad range of picornavirus RNAs, we generated a DNAzyme against a highly conserved sequence in the 5' untranslated region (5' UTR). While this DNAzyme was highly efficient against the 5' UTR of the human rhinovirus 14, it failed to cleave the identical target sequence within the RNA of the related coxsackievirus A21 (CAV-21). After introduction of 2'-O-methyl RNA or locked nucleic acid (LNA) monomers into the substrate recognition arms, the DNAzyme degraded the previously inaccessible virus RNA at a high catalytic rate even to completion, indicating that nucleotides with high target affinity were able to compete successfully with internal structures. We then adopted this strategy to two DNAzymes that we had found to be inactive in our earlier experiments. The modified DNAzymes proved to be highly effective against their respective target structures. Our approach may be useful for other ribozyme strategies struggling with accessibility problems, especially when being restricted to unique target sites. PMID- 15136039 TI - Recombination of hybrid target sites by binary combinations of Flp variants: mutations that foster interprotomer collaboration and enlarge substrate tolerance. AB - Strategies of directed evolution and combinatorial mutagenesis applied to the Flp site-specific recombinase have yielded recombination systems that utilize bi specific hybrid target sites. A hybrid site is assembled from two half-sites, each harboring a distinct binding specificity. Satisfying the two specificities by a binary combination of Flp variants, while necessary, may not be sufficient to elicit recombination. We have identified amino acid substitutions that foster interprotomer collaboration between partner Flp variants to potentiate strand exchange in hybrid sites. One such substitution, A35T, acts specifically in cis with one of the two partners of a variant pair, Flp(K82M) and Flp(A35T, R281V). The same A35T mutation is also present within a group of mutations that rescue a Flp variant, Flp(Y60S), that is defective in establishing monomer-monomer interactions on the native Flp target site. Strikingly, these mutations are localized to peptide regions involved in interdomain and interprotomer interactions within the recombination complex. The same group of mutations, when transferred to the context of wild-type Flp, can relax its specificity to include non-native target sites. The hybrid Flp systems described here mimic the naturally occurring XerC/XerD recombination system that utilizes two recombinases with distinct DNA binding specificities. The ability to overcome the constraints of binding site symmetry in Flp recombination has important implications in the targeted manipulations of genomes. PMID- 15136040 TI - The chromosome of Shigella flexneri bacteriophage Sf6: complete nucleotide sequence, genetic mosaicism, and DNA packaging. AB - Shigella flexneri temperate bacteriophage Sf6 is of interest in part because its prophage expresses the oac gene that alters the antigenic properties of the surface O-antigen polysaccharide of its host bacterium. We have determined the complete sequence of its 39,044 bp genome. The sequence shows that Sf6 is a member of the canonical lambdoid phage group, and like other phages of this type has a highly mosaic genome. It has chromosomal regions that encode proteins >80% identical with at least 15 different previously characterized lambdoid phages and prophages, but 43% of the genome, including the virion assembly genes, is homologous to the genome of one phage, HK620. An analysis of the nucleotide differences between Sf6 and HK620 indicates that even these similar regions are highly mosaic. This mosaicism suggests ways in which the virion structural proteins might interact with each other. The Sf6 early operons are arranged like a typical lambdoid phage, with "boundary sequences" often found between functional modules in the "metabolic" genome domain. By virtue of high degree of similarity in the encoding genes and their DNA target sites, we predict that the integrase, early transcription anti-terminator, CI and Cro repressors, and CII protein of Sf6 have DNA binding specificities very similar to the homologous proteins encoded by phages HK620, lambda, 434 and P22, respectively. The late operon contains two tRNA genes. The Sf6 terminase genes are unusual. Analysis of in vivo initiation of the DNA packaging series showed that the Sf6 apparatus that recognizes DNA for packaging appears to cleave DNA for initiation of packaging series at many sites within a large region of about 1800 bp that includes a possible pac site. This is unlike previously characterized phage packaging mechanisms. PMID- 15136041 TI - De novo DNA synthesis by human DNA polymerase lambda, DNA polymerase mu and terminal deoxyribonucleotidyl transferase. AB - DNA polymerases (pols) catalyse the synthesis of DNA. This reaction requires a primer-template DNA in order to grow from the 3'OH end of the primer along the template. On the other hand terminal deoxyribonucleotidyl transferase (TdT) catalyses the addition of nucleotides at the 3'OH end of a DNA strand, without the need of a template. Pol lambda and pol micro are ubiquitous enzymes, possess both DNA polymerase and terminal deoxyribonucleotidyl transferase activities and belong to pol X family, together with pol beta and TdT. Here we show that pol lambda, pol micro and TdT, all possess the ability to synthesise in vitro short fragments of DNA in the absence of a primer-template or even a primer or a template in the reaction. The DNA synthesised de novo by pol lambda, pol micro and TdT appears to have an unusual structure. Furthermore we found that the amino acid Phe506 of pol lambda is essential for the de novo synthesis. This novel catalytic activity might be related to the proposed functions of these three pol X family members in DNA repair and DNA recombination. PMID- 15136042 TI - Fitness increase of memory genomes in a viral quasispecies. AB - Viral quasispecies may contain a subset of minority genomes that reflect those genomic sequences that were dominant at an early phase of quasispecies evolution. Such minority genomes are referred to as memory in viral quasispecies. A memory marker previously characterized in foot-and-mouth disease virus (FMDV) is an internal oligoadenylate tract of variable length that became dominant upon serial plaque-to-plaque transfers of FMDV clones. During large population passages, genomes with internal oligoadenylate were outcompeted by wild-type revertants but remained in the mutant spectra as memory genomes. Here, we report a quantification of relative fitness of several FMDV clones, harboring internal oligoadenylate tracts of different length, and that were retrieved at early or late times (passage number) after implementation of memory. The results show that for any given length range of the oligoadenylate, maintenance in memory resulted in an increase in relative fitness, comparable to the increase undergone by the entire population. The fitness increase is in agreement with the Red Queen hypothesis, and implies a replicative memory mechanism. Thus, permanence of memory genomes may be a source of high fitness variants despite their initial low fitness, and despite having remained hidden in mutant spectra. This reinforces the interest of diagnosing minority genomes during chronic human and animal viral infections. PMID- 15136043 TI - Crystal structure of alpha-galactosidase from Trichoderma reesei and its complex with galactose: implications for catalytic mechanism. AB - The crystal structures of alpha-galactosidase from the mesophilic fungus Trichoderma reesei and its complex with the competitive inhibitor, beta-d galactose, have been determined at 1.54 A and 2.0 A resolution, respectively. The alpha-galactosidase structure was solved by the quick cryo-soaking method using a single Cs derivative. The refined crystallographic model of the alpha galactosidase consists of two domains, an N-terminal catalytic domain of the (beta/alpha)8 barrel topology and a C-terminal domain which is formed by an antiparallel beta-structure. The protein contains four N-glycosylation sites located in the catalytic domain. Some of the oligosaccharides were found to participate in inter-domain contacts. The galactose molecule binds to the active site pocket located in the center of the barrel of the catalytic domain. Analysis of the alpha-galactosidase- galactose complex reveals the residues of the active site and offers a structural basis for identification of the putative mechanism of the enzymatic reaction. The structure of the alpha-galactosidase closely resembles those of the glycoside hydrolase family 27. The conservation of two catalytic Asp residues, identified for this family, is consistent with a double displacement reaction mechanism for the alpha-galactosidase. Modeling of possible substrates into the active site reveals specific hydrogen bonds and hydrophobic interactions that could explain peculiarities of the enzyme kinetics. PMID- 15136044 TI - In vitro characterization of FlgB, FlgC, FlgF, FlgG, and FliE, flagellar basal body proteins of Salmonella. AB - The bacterial flagellar basal body is a rotary motor. It spans the cytoplasmic and outer membranes and drives rapid rotation of a long helical filament in the cell exterior. The flagellar rod at its central axis is a drive shaft that transmits torque through the hook to the filament to propel the bacterial locomotion. To study the structure of the rod in detail, we have established purification procedures for Salmonella rod proteins, FlgB, FlgC, FlgF, FlgG, and also for FliE, a rod adapter protein, from an Escherichia coli expression system. While FlgF was highly soluble, FlgB, FlgC, FlgG and FliE tended to self or cross aggregate into fibrils in solutions at neutral pH or below, at high ionic strength, or at high protein concentration. These aggregates were characterized to be beta-amyloid fibrils, unrelated to the rod structure formed in vivo. Under non-aggregative conditions, no protein-protein interactions were detected between any pairs of these five proteins, suggesting that their spontaneous, template free polymerization is strongly suppressed. Limited proteolyses showed that FlgF and FlgG have natively unfolded N and C-terminal regions of about 100 residues in total just as flagellin does, whereas FlgB, FlgC and FliE, which are little over 100 residues long, are unfolded in their entire peptide chains. These results together with other data indicate that all of the ten flagellar axial proteins share structural characteristics and folding dynamics in relation to the mechanism of their self-assembly into the flagellar axial structure. PMID- 15136045 TI - Crystal structure of a heat-resilient phytase from Aspergillus fumigatus, carrying a phosphorylated histidine. AB - In order to understand the structural basis for the high thermostability of phytase from Aspergillus fumigatus, its crystal structure was determined at 1.5 A resolution. The overall fold resembles the structure of other phytase enzymes. Aspergillus niger phytase shares 66% sequence identity, however, it is much less heat-resistant. A superimposition of these two structures reveals some significant differences. In particular, substitutions with polar residues appear to remove repulsive ion pair interactions and instead form hydrogen bond interactions, which stabilize the enzyme; the formation of a C-terminal helical capping, induced by arginine residue substitutions also appears to be critical for the enzyme's ability to refold to its active form after denaturation at high temperature. The heat-resilient property of A.fumigatus phytase could be due to the improved stability of regions that are critical for the refolding of the protein; and a heat-resistant A.niger phytase may be achieved by mutating certain critical residues with the equivalent residues in A.fumigatus phytase. Six predicted N-glycosylation sites were observed to be glycosylated from the experimental electron density. Furthermore, the enzyme's catalytic residue His59 was found to be partly phosphorylated and thus showed a reaction intermediate, providing structural insight, which may help understand the catalytic mechanism of the acid phosphatase family. The trap of this catalytic intermediate confirms the two-step catalytic mechanism of the acid histidine phosphatase family. PMID- 15136046 TI - Mutational analysis of the energetics of the GrpE.DnaK binding interface: equilibrium association constants by sedimentation velocity analytical ultracentrifugation. AB - DnaK, the prokaryotic Hsp70 molecular chaperone, requires the nucleotide exchange factor and heat shock protein GrpE to release ADP. GrpE and DnaK are tightly associated molecules with an extensive protein-protein interface, and in the absence of ADP, the dissociation constant for GrpE and DnaK is in the low nanomolar range. GrpE reduces the affinity of DnaK for ADP, and the reciprocal linkage is also true: ADP reduces the affinity of DnaK for GrpE. The energetic contributions of GrpE side-chains to GrpE-DnaK binding were probed by alanine scanning mutagenesis. Sedimentation velocity (SV) analytical ultracentrifugation (AUC) was used to measure the equilibrium constants (Keq) for GrpE binding to the ATPase domain of DnaK in the presence of ADP. ADP-bound DnaK is the natural target of GrpE, and the addition of ADP (final concentration of 5 microM) to the preformed GrpE-DnaK(ATPase) complexes allowed the equilibrium association constants to be brought into an experimentally accessible range. Under these experimental conditions, the substitution of one single GrpE amino acid residue, arginine 183 with alanine, resulted in a GrpE-DnaK(ATPase) complex that was weakly associated (Keq =9.4 x 10(4) M). This residue has been previously shown to be part of a thermodynamic linkage between two structural domains of GrpE: the thermosensing long helices and the C-terminal beta-domains. Several other GrpE side-chains were found to have a significant change in the free energy of binding (DeltaDeltaG approximately 1.5 to 1.7 kcal mol(-1)), compared to wild-type GrpE.DnaK(ATPase) in the same experimental conditions. Overall, the strong interactions between GrpE and DnaK appear to be dominated by electrostatics, not unlike barnase and barstar, another well-characterized protein-protein interaction. GrpE, an inherent thermosensor, exhibits non-Arrhenius behavior with respect to its nucleotide exchange function at bacterial heat shock temperatures, and mutation of several solvent-exposed side-chains located along the thermosensing indicated that these residues are indeed important for GrpE-DnaK interactions. PMID- 15136047 TI - Sensor domain of the Mycobacterium tuberculosis receptor Ser/Thr protein kinase, PknD, forms a highly symmetric beta propeller. AB - Diverse pathogenic bacteria produce transmembrane receptor Ser/Thr protein kinases (STPKs), but little is known about the signals mediated by these "eukaryotic-like" proteins. To explore the basis for signaling in the bacterial STPK receptor family, we determined the structure of the sensor domain of Mycobacterium tuberculosis PknD. In two crystal forms, the PknD sensor domain forms a rigid, six-bladed beta-propeller with a flexible tether to the transmembrane domain. The PknD sensor domain is the most symmetric beta-propeller structure described. All residues that vary most among the blade subdomains cluster in the large "cup" motif, analogous to the ligand-binding surface in many beta-propeller proteins. These results suggest that PknD binds a multivalent ligand that signals by changing the quaternary structure of the intracellular kinase domain. PMID- 15136048 TI - Snorkeling preferences foster an amino acid composition bias in transmembrane helices. AB - By analyzing transmembrane (TM) helices in known structures, we find that some polar amino acids are more frequent at the N terminus than at the C terminus. We propose the asymmetry occurs because most polar amino acids are better able to snorkel their polar atoms away from the membrane core at the N terminus than at the C terminus. Two findings lead us to this proposition: (1) side-chain conformations are influenced strongly by the N or C-terminal position of the amino acid in the bilayer, and (2) the favored snorkeling direction of an amino acid correlates well with its N to C-terminal composition bias. Our results suggest that TM helix predictions should incorporate an N to C-terminal composition bias, that rotamer preferences of TM side-chains are position dependent, and that the ability to snorkel influences the evolutionary selection of amino acids for the helix N and C termini. PMID- 15136049 TI - Approaches to measuring cholesterol absorption in humans. AB - Under optimal conditions, plasma cholesterol homeostasis is maintained by a variety of mechanisms, balancing input and output, thereby preventing the net accumulation of cholesterol in circulation and tissues. Among these mechanisms, intestinal cholesterol absorption has recently re-emerged as a potentially important contributor to cholesterol homeostasis. However, its regulation has been difficult to study in humans because of technical limitations in methodologies. In this review the major methods available for measuring cholesterol absorption including those that utilize cholesterol balance, single dose isotopic feeding, dual isotope plasma ratio, continuous isotope feeding, intestinal perfusion, stable isotopes and serum plant sterols or cholestanol to cholesterol ratios are reviewed and contrasted. Emphasis is placed on the strengths, technical and interpretational limitations and their applicability for use in metabolic, small-scale outpatient, population and large-scale intervention studies. PMID- 15136050 TI - Aspirin inhibits monocyte chemoattractant protein-1 and interleukin-8 expression in TNF-alpha stimulated human umbilical vein endothelial cells. AB - Atherosclerosis and its complications such as stroke, myocardial infraction and peripheral vascular disease, remain the major causes of morbidity and mortality in the world. Studies have showed that chemokines and adhesion molecules are involved in causing atherosclerosis by promoting directed migration of inflammatory cells. Monocyte chemoattractant protein-1 (MCP-1) is one of the key factors critical for the initiating and developing of atherosclerotic lesions. IL 8, a CXC chemokine, stimulates neutrophil chemotaxis. Aspirin is the most common drug used to prevent the complications of atherosclerosis such as stroke and coronary heart disease. In this study, we found that aspirin inhibited TNF-alpha (10 ng/ml)-induced MCP-1 and IL-8 expression at the RNA and protein levels in human umbilical vein endothelial cells (HUVECs), monocyte adhesion and transmigration, and that its inhibitory effects were not due to decreased HUVEC viability as assessed by MTT test. Aspirin at the dose as low as 10 microg/ml significantly inhibited the release of TNF-stimulated MCP-1 by 29.1% (P = 0.008) and IL-8 by 26.9% (P = 0.0146) as compared to TNF-stimulated release. Antibodies pretreatment were likely to decrease the production of MCP-1 (P < 0.0001) and IL 8 (P < 0.0001). Furthermore, aspirin (10 microg/ml) inhibited U937 cell adhesion by a 13.4% (P = 0.0119) inhibition as compared to TNF-stimulated alone. Finally, at higher concentration, aspirin also inhibited U937 migration to HUVEC by 89.1% (P = 0.0475) as compared to TNF-stimulated alone. These results in our study suggest that aspirin inhibits TNF-alpha stimulated MCP-1 and IL-8 release in HUVECs, for its additional therapeutic effects of aspirin in causing atherosclerosis. PMID- 15136051 TI - Inhibitory effects of tetrandrine on the serum- and platelet-derived growth factor-BB-induced proliferation of rat aortic smooth muscle cells through inhibition of cell cycle progression, DNA synthesis, ERK1/2 activation and c-fos expression. AB - Tetrandrine (TET) is a well known naturally occurred nonspecific Ca(2+) channel blocker. It has long been used for the treatment of arrhythmia, hypertension, and occlusive cardiovascular disorders. The objective of the present study was to investigate the effect of TET on the proliferation of primary cultured rat aortic smooth muscle cells (RASMCs). TET significantly inhibited both 10% fetal bovine serum (FBS) and 50 ng/ml platelet-derived growth factor (PDGF)-BB-induced proliferation, [(3) H] ]thymidine incorporation into DNA, and p42/p44 mitogen activated protein kinase (ERK1/2) phosphorylation at the concentration of 1.0 and 5.0 microM. Flow cytometry analysis of DNA content in synchronized cells revealed blocking of the FBS-inducible cell cycle progression by TET. In accordance with these findings, TET 5 microM caused a 48% decrease in the early elevation of c fos expression induced after 10% FBS addition. Furthermore, in contrast to its distinguishable higher potency of Ca(2+) antagonistic activity, verapamil showed lower potent antiproliferative activities than TET. These results suggest that TET can exert antiproliferative effects against mitogenic stimuli for RASMCs in vitro by a mechanism that involves the MAPK pathway, altering cell cycle progression, and the inhibitory action cannot be limited to its Ca(2+) modulation. PMID- 15136052 TI - Interrelationships between the fibrinolytic system and lipoproteins in the pathogenesis of coronary atherosclerosis. AB - The fibrinolytic system is comprised of a series of serine proteases and serine protease inhibitors which are involved in the dissolution of fibrin in the vascular lumen, but also in the migration of cells and in the remodeling of the extracellular matrix of the vascular wall. The transcription, expression and degradation of the various fibrinolytic enzymes by cells in the vascular wall is influenced by lipoproteins and this interrelationship may play a significant role in the development of the atherosclerotic plaque: the transcription of plasminogen activator inhibitor-1 is influenced by very low-density lipoproteins, the expression of both tissue plasminogen activator and plasminogen activator inhibitor-1 is influenced by low-density lipoproteins and lipoprotein(a) (Lp(a)) and the internalization of the urokinase: plasminogen activator inhibitor-1 complex occurs via the low-density lipoprotein related protein. Several clinical studies have shown correlations between fibrinolytic parameters and lipoproteins in healthy populations and in patients with dyslipidemia, but the correlation between single plasma fibrinolytic enzymes and the severity of coronary atherosclerosis is less well documented. The reduction of plasma lipids with lipid-lowering drugs also affects the concentration of fibrinolytic enzymes, although this may also be due to direct effects of the drugs on the expression of the various fibrinolytic enzymes. The reduction of fibrinolytic and proteolytic activity in the atherosclerotic plaque by their lipid-lowering effect and by their direct action on the fibrinolytic system may be one of the mechanisms by which some lipid-lowering drugs achieve plaque stabilization. PMID- 15136053 TI - Rosiglitazone improves intestinal lipoprotein overproduction in the fat-fed Syrian Golden hamster, an animal model of nutritionally-induced insulin resistance. AB - We have recently shown that the fructose-fed Syrian Golden hamster, a non diabetic animal model of nutritionally-induced insulin resistance and hyperlipidemia, is characterized by intestinal lipoprotein overproduction. In order to determine whether intestinal lipoprotein overproduction is specific to fructose feeding or applies generally to other models of insulin resistance, we studied intestinal lipoprotein production and the response to insulin sensitization in the high fat-fed Syrian Golden hamster. Syrian Golden Hamsters were fed either (1). chow (CHOW), (2). 60% fat (FAT) or (3). 60% fat with rosiglitazone, 20 micromol/kg per day (FAT + RSG) for 5 weeks. Euglycemic hyperinsulinemic clamp studies confirmed that FAT is a good model of insulin resistance and rosiglitazone treatment resulted in a significant improvement in insulin sensitivity. In addition, there was a significant approx. two- to four fold increase in intestinal apoB48 particle production in FAT. Rosiglitazone treatment resulted in partial normalization of apoB48-containing intestinal lipoprotein secretion. In summary: (1). the fat-fed Syrian Golden Hamster is a good model of nutritionally-induced insulin resistance, (2). intestinal overproduction of lipoproteins appear to contribute to the hypertriglyceridemia of insulin resistance in this animal model and (3). insulin sensitization with rosiglitazone ameliorates intestinal apoB48 particle overproduction in the fat fed Syrian Golden Hamster. These data further support the link between insulin resistance and intestinal lipoprotein overproduction. PMID- 15136054 TI - Multidetector-row computed tomography and magnetic resonance imaging of atherosclerotic lesions in human ex vivo coronary arteries. AB - In the present study, we tested the ability of multidetector-row computed tomography (MDCT) and magnetic resonance imaging (MRI) to identify and retrospectively characterize atherosclerotic lesions in human ex vivo coronary arteries. Thirteen ex vivo hearts were studied with MDCT and MRI. MDCT-images were obtained with an isotropic voxel size of 0.6mm(3). MR images were obtained with an in-plane resolution of 195 microm and 3mm slice thickness. All images were matched with histopathology sections. For both modalities, the sensitivity for the detection of any atherosclerotic lesion was evaluated, and a retrospective analysis of plaque morphology according to criteria defined by the American Heart Association (AHA) was performed. At histopathology, 28 atherosclerotic lesions were found. 21 and 23 of these lesions were identified by MDCT and MRI, respectively. Both modalities detected a small number of false positive lesions. After retrospective matching with histopathology, MDCT as well as MRI were able to differentiate typical morpholocigal features for fatty, fibrous or calcified plaque components. Using the information presented in this study, in vivo coronary artery wall imaging using MDCT as well as MRI could be facilitated and supported for future investigations on this subject. PMID- 15136055 TI - Inhibitory efficacy of pitavastatin on the early inflammatory response and neointimal thickening in a porcine coronary after stenting. AB - Neointimal hyperplasia plays a crucial role in restenosis after stenting. The severity of neointimal thickness correlates with inflammatory reactions in the injured vessel and statins can inhibit inflammation. Pitavastatin has favorable effects on plasma lipoproteins and inflammation. Thus, we hypothesized that pitavastatin might inhibit the early inflammatory response, resulting in prevention of neointimal hyperplasia in porcine coronary arteries after stenting. Pitavastatin (18 coronaries, 40 mg per day) or placebo (20 coronaries) was administered orally from 7 days before stenting until the time of euthanasia at 3 or 28 days after stenting. The coronary artery of the animals was injured with an oversized metallic coil stent. Inflammatory cell infiltration was evaluated by scanning electron microscopy and was significantly reduced in the treated vessels compared to controls. On Day 28, intravascular ultrasound analysis revealed the neointimal area was significantly less at the stent site in the pitavastatin group than in the placebo. Histopathologic assessment showed significantly decreased in neointimal area in the pitavastatin group compared to the placebo (2.16 +/- 0.13 mm(2) versus 2.88 +/- 0.25 mm(2), p = 0.029), whereas the mean injury score in the pitavastatin group was larger than in the placebo group. In conclusion, Pitavastatin inhibited neointimal hyperplasia after stenting through a reduction of inflammatory reactions. PMID- 15136056 TI - Conjugated linoleic acid and atherosclerosis: no effect on molecular markers of cholesterol homeostasis in THP-1 macrophages. AB - Macrophage cholesterol homeostasis is a key process involved in the initiation and progression of atherosclerosis. Peroxisome proliferator-activated receptors (PPARs) regulate the transcription of the genes involved in cholesterol homeostasis and thus represent an important therapeutic target in terms of reducing atherosclerosis. Conjugated linoleic acid (CLA) is a potent anti atherogenic dietary fatty acid in animal models of atherosclerosis and is capable of activating PPARs in vitro and in vivo. Therefore, this study examined whether the anti-atherogenic effects of CLA in vivo could be ascribed to altered cholesterol homeostasis in macrophages and macrophage derived foam cells. Of several genes that regulate cholesterol homeostasis investigated, CLA had most effect on the class B scavenger receptor CD36. The cis-9,trans-11 CLA (c9,t11 CLA) and trans-10,cis-12 CLA (t10,c12-CLA) isomers augmented CD36 mRNA expression (P<0.001). Confocal laser microscopy characterised the three-dimensional expression patterns of CD36 in THP-1 macrophages. Linoleic acid, CLA and the PPARgamma ligand rosiglitazone increased discrete cell surface CD36 localisation, with a heterogeneous punctate pattern of expression. In agreement with the observed increases in CD36 mRNA and cell surface expression, intracellular cholesterol concentrations were greater in macrophages exposed to linoleic acid and CLA. Further analysis of cholesterol metabolism showed that CLA had no effect on THP-1 derived foam cell cholesterol efflux to apo AI. Thus, altered cholesterol homeostasis in the macrophage may not explain the anti-atherogenic effects of CLA observed in vivo. PMID- 15136057 TI - Phenytoin treatment reduces atherosclerosis in mice through mechanisms independent of plasma HDL-cholesterol concentration. AB - Phenytoin (PHT) increases high density lipoprotein cholesterol (HDL-C) and reduces coronary artery disease mortality in humans. We report the results of PHT treatment on atherosclerosis susceptibility and lipid profile in four different types of mouse: control C57BL/6 mice and cholesteryl ester transfer protein transgenic mice as models of fatty streak, and LDL receptor-deficient mice and apolipoprotein E-deficient mice as models of mature atherosclerosis. Each mouse type was fed an appropriate diet to induce atherosclerosis and prevent liver toxicity. PHT treatment demonstrated a protective effect in all models. Reduction in aortic atherosclerotic area by PHT treatment was more evident in early atherosclerosis (2.3-fold) than in mature atherosclerosis (decreases of 40 and 23%, respectively, but only in mice in the upper 50% percentile of plasma PHT concentration). Atherosclerosis prevention was not concomitant with a consistent increase in HDL-C or any other protective change in the lipid profile. Different analyses of potential antiatherogenic HDL functions did not provide additional information. Microarray liver gene expression analyses identified a potential atheroprotective mechanism characterized by decreased expression of syndecan-4, RhoA2, double LIM protein-1, zeta-chain-associated protein kinase-70 and interleukin 6 receptor-alpha. However, to demonstrate that these changes are part of a PHT-antiatherogenic effect, they will need to be found also in arteries, maintained at protein level and proved to be causal rather than reactive. PMID- 15136058 TI - N(omega)-(carboxymethyl)lysine depositions in human aortic heart valves: similarities with atherosclerotic blood vessels. AB - Recent studies indicate a role of atherosclerosis-like changes involved in the pathogenesis of aortic valve stenosis. Interestingly, one of the major advanced glycation end products (AGEs), N(omega)-(carboxymethyl)lysine (CML) has been related to the process of atherosclerosis in blood vessels. In the present study, we have analyzed the presence of CML in degenerative altered aortic valves with atherosclerosis-like changes, and in degenerated mitral valves without atherosclerosis-like changes, derived from patients suffering from acute rheumatism during childhood. Degenerated and non-degenerated valves were derived from autopsy or obtained during cardiac surgery. The presence of CML was examined by immunohistochemistry. CML was found on the endothelium and fibroblasts in control aortic and mitral valves. Minor differences in CML staining were observed between control and degeneratively affected mitral valves. In contrast, in degenerated aortic valves, CML accumulation was found in macrophages and on calcification sites, comparable to that in atherosclerotic arteries, while the presence of CML staining on the endothelium and fibroblasts was significantly less as compared with control aortic valves. Our data support the hypothesis that the process of degeneration of aortic valves resembles that of atherosclerosis in blood vessels. They suggest that CML also plays a role in the process of atherosclerosis in aortic valves. PMID- 15136059 TI - Antiatherogenic effect of Pistacia lentiscus via GSH restoration and downregulation of CD36 mRNA expression. AB - Pistacia lentiscus var. Chia (Anacardiaceae) grows almost exclusively on Chios Island, Greece, and gives a resinous exudate resin used for culinary purposes by Mediterranean people. We investigated the molecular mechanisms through which total polar extract of the resin inhibits oxidized low-density lipoprotein (oxLDL) cytotoxic effect on peripheral blood mononuclear cell (PBMC). Cells exposed to oxLDL underwent apoptosis and necrosis, dependent on the duration of exposure. When culturing cells with oxLDL and the polar extract concurrently, we observed inhibition of both the phenomena. Because under oxidative stress the pro oxidant systems outbalance the antioxidant, potentially producing oxidative damage and ultimately leading to cell death, we measured the levels of intracellular antioxidant glutathione (GSH). Additionally, we measured CD36 expression, a class B scavenger receptor, on CD14-positive cells, as CD36 has been identified as the oxLDL receptor in macrophages and may play a pivotal role in atherosclerotic foam cell formation. oxLDL decreased GSH levels and upregulated CD36 expression. P. lentiscus extract restored GSH levels and downregulated CD36 expression, even at the mRNA level. In order to find out the biologically drastic constituents of the resin's polar extract, fractions derived from RP-HPLC analysis were examined for their antioxidant effect on oxidatively stressed PBMC. The triterpenoid fraction revealed remarkable increase in intracellular GSH. We suggest GSH restoration and downregulation of CD36 mRNA expression as the pathways via which P. lentiscus triterpenes exert antioxidant/antiatherogenic effect. Additionally, our results provide strong evidence of the resin's antiatherogenic effect; therefore it is credited with beneficial health aspects. PMID- 15136060 TI - Simvastatin potenciates PGI(2) release induced by HDL in human VSMC: effect on Cox-2 up-regulation and MAPK signalling pathways activated by HDL. AB - High density lipoproteins (HDL) induce prostacyclin (PGI(2)) release in vascular smooth muscle cells (VSMC) by up-regulation of cyclooxygenase-2 (Cox-2). Our goal was to analyse the mechanisms underlying this effect, and its potential modulation by HMG-CoA reductase inhibition in human VSMC. The contribution of mitogen-activated protein kinase (MAPK) signalling pathways was assessed by Western blot analysis and using specific inhibitors [PD098059 for p42/44 MAPK kinase (MEK); SB203580 for p38 MAPK or L-JNKI1 for c-Jun N-terminal kinase-1 (JNK 1)]. HDL-induced PGI(2) release was inhibited by rofecoxib (a specific Cox-2 inhibitor, 5 microM). HDL induced the early activation of p42 MAPK, p38 MAPK and JNK-1. p42/44 MAPK was the major pathway involved in both Cox-2 up-regulation and PGI(2) synthesis; p38 MAPK was also involved in both processes while JNK inhibition only affected PGI(2) synthesis. Pertussis toxin (an inhibitor of Galphai/Galphao proteins) prevented MAPK activation and inhibited both Cox-2 up regulation and PGI(2) release. Genistein (a tyrosine kinase inhibitor) inhibited PGI(2) release without affecting MAPK activation or Cox-2 up-regulation. Simvastatin (0.1-1 microM) increased HDL-induced PGI(2) release ( approximately 45% at 1 microM) but did not significantly modify early MAPK activation or Cox-2 expression. Simvastatin alone did not significantly affect PGI(2) release. Our results suggest that mechanisms associated with G protein-coupled receptor activation, trigger Cox-2 up-regulation and PGI(2) release via multiple MAPK signalling pathways in VSMC. The mechanism is independent of tyrosine kinase receptors, although cytosolic tyrosine kinases could activate Cox-2 post translationally. The potential contribution of HDL to vascular homeostasis, via increases in PGI(2) synthesis, could be enhanced by HMG-CoA reductase inhibitors. PMID- 15136061 TI - The 5,10-methylenetetrahydrofolate reductase C677T polymorphism interacts with smoking to increase homocysteine. AB - Elevated homocysteine is a risk marker for several human pathologies. Risk factors for elevated homocysteine include low folate and homozygosity for the T allele of the 5,10-methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism. Because nitric oxide may inhibit folate catabolism and endothelial nitric oxide synthase activity is reduced in smokers, we postulated that smoking status might modify the impact of the MTHFR C677T polymorphism on homocysteine (tHcy) concentrations. We tested this hypothesis in a healthy young adult population for which MTHFR C677T genotypes and tHcy concentrations were previously reported. The MTHFR 677TT genotype was significantly associated with elevated tHcy concentrations in smokers (P = 0.001) but not in non-smokers (P = 0.36). Among smokers, the MTHFR 677TT genotype was significantly associated with high tHcy in heavy smokers (P = 0.003) but not light smokers (P = 0.09), in men (P = 0.003) but not women (P = 0.11), and in subjects from the lowest serum folate quartile (P = 0.49) but not from folate quartiles 2-4 (P = 0.49). After adjustment for nutritional variables, interactions between MTHFR C677T genotype and NOS3 G894T genotype, and between MTHFR genotype, smoking status and gender were statistically significant. We propose that hyperhomocysteinemia in MTHFR 677TT homozygote smokers is the consequence of mild intracellular folate deficiency caused by a smoking-related reduction of NOS3 activity that is exacerbated when serum folate is low. PMID- 15136062 TI - L-arginine attenuates lymphocyte activation and anti-oxidized LDL antibody levels in patients undergoing angioplasty. AB - BACKGROUND: Patients with acute coronary syndromes exhibit evidence of peripheral T lymphocyte activation, elevated acute phase proteins and enhanced oxidative stress. Nitric oxide (NO) has been recognized as one of the relaxant factors synthesized and released by normal endothelium, and acts as a double-edged sword on the immune system. L-arginine ameliorates experimental atherosclerosis and restenosis as well as endothelial dysfunction. We sought to investigate the effect of L-arginine administration on the extent of lymphocyte activation and anti-oxLDL antibodies in patients with unstable angina undergoing PCI with stent placement. METHODS: Patients with unstable angina were randomized to treatment with L-arginine (6g per day; n = 13) or none (n = 16) for 1 month starting immediately on the day of stent deployment. Lymphocyte activation was assayed by FACS employing double staining with a common lymphocyte marker (CD3) and an activation marker HLA-DR, on the day of the procedure and 1 month later. Anti oxLDL antibodies were assayed by ELISA. RESULTS: Patients with unstable angina not receiving L-arginine exhibited a significant 43% rise in the percentage of activated peripheral T lymphocytes, 1 month after stent deployment. Patients treated with L-arginine exhibited a fall albeit not significant in the fraction of peripheral lymphocytes bearing the activation marker. Antibodies to anti-oxLDL rose significantly between baseline and 1 month follow-up. L-arginine treatment significantly attenuated the rise in anti-oxLDL antibody levels. CONCLUSION: L arginine attenuates the systemic rise in peripheral lymphocyte activation and oxidative stress markers induced by vessel wall injury following PCI. These effects may contribute to a favorable effect of the drug in patients with acute coronary syndromes undergoing PCI. PMID- 15136063 TI - Probucol and ticlopidine: effect on platelet and monocyte activation markers in hyperlipidemic patients with and without type 2 diabetes. AB - We investigated the effects of probucol and ticlopidine on circulating levels of platelet activation markers, microparticles, soluble selectins, and malondialdehyde-low density lipoprotein (MDA-LDL) in hyperlipidemic patients with or without type 2 diabetes. There were significant differences in the levels of CD62P, PAC-1, annexin V, PDMP, MDMP, sP-selectin, sE-selectin and MDA-LDL between the hyperlipidemic patients and the controls. In particular, these markers were significantly increased in hyperlipidemic patients who had type 2 diabetes. In the hyperlipidemic patients with diabetes, MDA-LDL was decreased by both monotherapy with probucol and combination therapy (probucol and ticlopidine). In these patients, CD62P, PAC-1, annexin V, MDMP, PDMP, sP-selectin, and sE-selectin were also significantly decreased after treatment. The decreases of CD62P, PAC-1, annexin V, PDMP and sP-selectin were greater combination therapy than with monotherapy. These findings suggest that administration of probucol and ticlopidine to hyperlipidemic patients with type 2 diabetes may help to prevent the development of cardiovascular complications caused by modified LDL, selectins, or activated platelets and monocytes. PMID- 15136064 TI - Inflammation, infection, and aortic valve sclerosis; Insights from the Olmsted County (Minnesota) population. AB - Atherosclerosis-related mechanisms, including inflammation and possibly infection, are likely to be involved in the pathogenesis of calcific aortic valve disease. The purpose of this study was to examine whether systemic inflammatory markers and Chlamydia pneumoniae seropositivity are associated with aortic valve sclerosis (AVS) in a sample of the general population. Transesophageal echocardiography was performed in 381 subjects (median age: 67 years, range: 51 101; 52% men), a sample of the adult population in Olmsted County, Minnesota. The associations between systemic inflammatory markers (blood counts, including white blood cells differential counts, fibrinogen, and high-sensitivity C-reactive protein [hs-CRP]), C. pneumoniae immunoglobulin G (IgG) antibody titers, and AVS were examined. AVS was present in 140 subjects (37% of the population). After adjustment for age, sex, and smoking status: (1). hs-CRP was associated with AVS (odds ratio: 1.20 per two-fold increase in hs-CRP; 95% confidence interval: 1.01 1.43; P = 0.04) but this association was not significant after adjustment for additional risk factors for AVS, including body mass index (P = 0.52). (2). Blood counts and fibrinogen were not associated with AVS (P-values >0.30). (3). C. pneumoniae IgG antibody titers (low [1:16-1:32], intermediate [1:64-1:128], or high [>or=1:256] titers, compared with titers <1:16) were not associated with AVS (P = 0.21). In conclusion, hs-CRP is weakly associated with AVS, an association that is not independent of other AVS risk factors. Blood counts, fibrinogen, and C. pneumoniae seropositivity are not associated with AVS. These findings suggest that other non-inflammatory non-infectious mechanisms are likely to have a role in the pathogenesis of calcific aortic valve disease. PMID- 15136065 TI - Circulating oxidized low-density lipoprotein is an independent predictor for cardiac event in patients with coronary artery disease. AB - Oxidized low-density lipoprotein (oxLDL) plays a crucial role in the development of atherosclerosis, however, the predictive value of circulating oxLDL for cardiac events (CE) in patients with coronary artery disease (CAD) has remained poorly understood. We prospectively studied 238 consecutive patients with documented CAD for up to 52 months until the occurrence of one of the following cardiac events: cardiac death, nonfatal myocardial infarction (MI), and refractory angina requiring revascularization. The plasma levels of oxLDL were measured by an enzyme-linked immunosorbent assay (ELISA) using the monoclonal antibody, DLH3. The levels of circulating oxLDL were significantly higher in patients with CE than in patients without CE (median 20.3 U/ml versus 17.6 U/ml, P = 0.002). Multivariate Cox models showed that higher level of oxLDL was an independent predictor of developing CE. The adjusted hazard ratios for CE were 3.15 (95% CI 1.47-6.76, P = 0.003) times higher in patients with the highest quartile of oxLDL levels and 1.88 (95% CI 0.90-3.95, P = 0.09) times higher in patients with the third quartile than in those within the lowest quartile. Thus, measurement of circulating oxLDL may be helpful in the assessment of future CE in patients with CAD. PMID- 15136066 TI - A serum amyloid A and LDL complex as a new prognostic marker in stable coronary artery disease. AB - BACKGROUND: Although some reports have indicated that acute phase proteins such as C-reactive protein (CRP) and serum amyloid A (SAA) can predict the prognosis in patients with acute coronary syndrome, the value of these markers in patients with stable coronary artery disease (CAD) still remains obscure. Therefore, our aim was to determine the prognostic value of inflammatory markers in patients with stable coronary artery disease. METHODS AND RESULTS: We conducted a prospective cohort study in 140 consecutive patients with stable coronary artery disease who had at least one coronary stenosis more than 50% in diameter seen on diagnostic coronary angiography (CAG). We determined serum levels of the SAA/LDL complex as a new marker in addition to CRP and SAA. Serum levels of the SAA/LDL complex were measured by a sandwich enzyme-linked immunosorbent assay (ELISA). End-points were defined as cardiac death, myocardial infarction, cerebral infarction, and coronary revascularization. End-point events occurred in 21 patients (2 death from myocardial infarction, 2 cerebral infarction, and 17 revascularization). Age (year) (OR = 1.14, CI: 1.05-1.25), diabetes mellitus (OR = 3.50, CI: 1.08-11.40), triglyceride (10mg/dl) (OR = 1.12, CI: 1.01-1.23) and SAA/LDL complex (10 microg/ml) (OR = 2.32, CI: 1.05-4.70) were independently related to the events. A reconstitution experiment suggested that the SAA/LDL complex is derived by oxidative interaction between SAA and lipoproteins. CONCLUSIONS: The SAA/LDL complex reflects intravascular inflammation directly and can be a new marker more sensitive than CRP or SAA for prediction of prognosis in patients with stable coronary artery disease. PMID- 15136067 TI - TXNIP gene not associated with familial combined hyperlipidemia in the NHLBI Family Heart Study. AB - Familial combined hyperlipidemia (FCHL) is the most common familial dyslipidemia, and is implicated in up to 20% of cases of premature coronary heart disease. Positive linkage to chromosome 1q was found in FCHL families participating in the NHLBI Family Heart Study (FHS), replicating linkage found in other studies. The HcB-19 mouse, which shares phenotypes with FCHL, was shown in other studies to have a nonsense mutation in the thioredoxin interacting protein gene (txnip). txnip is a gene on mouse chromosome 3 in a region syntenic with the 1q human FCHL linkage region. We re-sequenced the human homolog of mouse txnip in the FHS sample and identified nine single nucleotide polymorphisms (SNPs). We did not observe the nonsense mutation found in the HcB-19 mouse, and only three of the SNPs discovered were sufficiently polymorphic for analysis. No association between FCHL and the TXNIP gene was found. Within FCHL cases, presence of variants also did not significantly affect body mass index or levels of lipids, insulin, or glucose. Our results suggest that in this sample, TXNIP does not play a major role in FCHL or related traits, and is unlikely to account for the positive evidence of linkage in this region. PMID- 15136068 TI - Relationship of adiposity to the population distribution of plasma triglyceride concentrations in vigorously active men and women. AB - Although it is known that triglyceride concentrations increase with adiposity, whether the same increase applies for different percentiles of the triglyceride distribution has not been reported. Therefore, physician-supplied triglyceride concentrations from 7288 male and 2326 female runners were divided into strata according to the body mass index (BMI) and circumferences of the waist, hip and chest. The percentiles of the triglyceride distribution within each stratum were used to determine the cross-sectional regression slope between adiposity and triglyceride levels at each triglyceride percentile. Compared to the 5th percentile of the triglyceride distribution, the rise in men's triglycerides at the 95th percentile per unit of adiposity was 14-fold greater for BMI, 7.8-fold greater for waist circumference, 3.6-fold greater for hip circumference, and 4.4 fold greater for chest circumference. The rise in women's triglyceride concentrations at the 95th percentile was 8-fold greater than at the 5th percentile for each kg/m(2) increase in BMI. These results suggest that the metabolic effects of adiposity on plasma triglycerides depend upon whether the concentrations are high or low. This contradicts statistical assumptions upon which prior studies of adiposity have based their analyses. We speculate that the reported greater increases in triglycerides per unit of adiposity in whites than blacks, in men than women, and in low-density lipoprotein (LDL) pattern B than A are all consistent with the relationships we observe. It remains to be verified whether these relationship also apply to less active populations. PMID- 15136069 TI - The relationship between high-sensitive C-reactive protein and pulse wave velocity in healthy Japanese men. AB - Although pulse wave velocity (PWV) and high-sensitive C-reactive protein (hsCRP) are known as predictors of future cardiovascular events, their association has not been examined. The present study was conducted to evaluate their association in the general population. In 2668 Japanese men (43 +/- 10 years old), PWV was obtained by volume rendering methods, and hsCRP was determined by the latex aggregation method. PWV showed a significant correlation with logarithm of hsCRP (r = 0.06, P < 0.01). The concentration of hsCRP in the highest quartile of PWV was higher than that in the other three groups (P < 0.01). However, multiple linear regression analyses demonstrated that logarithm of hsCRP was not significantly related to PWV, independent from conventional risk factors. Calculated Framingham risk score (FRS) was higher in the highest quartiles of both hsCRP and PWV than in the other groups (P < 0.05). Thus, while increased hsCRP related to increased PWV, they may be independent predictors of atherosclerotic cardiovascular risk. A prospective study to confirm the independency of their significance in predicting future cardiovascular events and to evaluate the usefulness of the combination of both parameters to screen subjects for cardiovascular risk is necessary. PMID- 15136070 TI - Effects of fenofibrate on lipoproteins, vasomotor function, and serological markers of inflammation, plaque stabilization, and hemostasis. AB - We investigated the effects of fenofibrate, peroxisome proliferator-activated receptors (PPARs) agonist, on endothelial function in patients with hypertriglyceridemia. We administered placebo or fenofibrate 200 mg daily to 25 patients with hypertriglyceridemia for 8 weeks. This study was randomized, double blind, placebo-controlled, crossover in design. Compared with placebo, fenofibrate significantly changed lipoprotein levels including non-HDL cholesterol and significantly improved the percent flow-mediated dilator response to hyperemia by 13 +/- 6% (P < 0.001) and lowered plasma levels of tumor necrosis factor-alpha by 13 +/- 3% (P < 0.001). Fenofibrate reduced fibrinogen and plasminogen activator inhibitor type 1 antigen levels by 17 +/- 3 and 10 +/- 3%, respectively (P < 0.001 and P = 0.014, respectively). However, fenofibrate did not significantly change plasma levels of nitrate, malondialdehyde, tissue factor activity, and serological markers of plaque stabilization. Fenofibrate significantly changed lipoprotein levels and improved the percent flow-mediated dilator response to hyperemia as well as lowered levels of tumor necrosis factor alpha (TNF-alpha), fibrinogen, and plasminogen activator inhibitor type 1 antigen. PMID- 15136071 TI - Lipoprotein lipase (LPL) mass in preheparin serum reflects insulin sensitivity. AB - Lipoprotein lipase (LPL) is one of the enzymes regulated by insulin and its plasma activity reflects insulin sensitivity. Although intravenous heparin injection is required to measure LPL activity, we can detect LPL mass in preheparin serum (Pr-LPL mass) by immunoassay. In this study, we examined whether Pr-LPL mass reflects insulin sensitivity. We measured Pr-LPL mass, insulin sensitivity (Si), and acute insulin release in response to a glucose bolus (AIRg) in subjects with normal glucose tolerance (NGT; n = 23), impaired glucose tolerance (IGT; n = 10), and Type II diabetes mellitus (DM; n = 48). Si and AIRg were determined by minimal model analysis. We also compared Pr-LPL mass with the homeostasis model assessment of insulin resistance (HOMA-R) and the urinary excretion of C-peptide (urine CPR). We found that Pr-LPL mass correlated significantly with Si ( r = 0.354, P < 0.01) in all the subjects. This correlation was still significant in the NGT group (P < 0.472, P < 0.05), DM group (r = 0.311, P < 0.01), and DM group with a fasting plasma glucose >150 mg/dl ( n = 20, r = 0.459. P < 0.05). Moreover, Pr-LPL mass correlated negatively with HOMA-R (r = -0.272. P < 0.05) and fasting IRI (r = -0.256, P < 0.05). By contrast, Pr-LPL mass was not correlated with either urine CPR or logAIRg that reflect the ability to secrete insulin. In conclusion, Pr-LPL mass reflects insulin sensitivity. We speculate that Pr-LPL mass might be used to assess insulin sensitivity not only in the general population but also in advanced diabetic patients. PMID- 15136072 TI - Serum lipids of Greenland Inuit in relation to Inuit genetic heritage, westernisation and migration. AB - BACKGROUND: The reputed low prevalence of cardiovascular disease among the Inuit has recently been challenged. Studies have shown total cholesterol among the Inuit to differ little from that of western populations and the association between cholesterol and atherosclerosis to be inconsistent. METHODS: We studied serum lipids in a population survey among 2114 Inuit living in Denmark or in West Greenland. Blood tests were supplemented by structured interviews, anthropometry and measurements of blood pressure. FINDINGS: Compared with the general population of Denmark, total cholesterol was higher among Inuit women, while HDL cholesterol was higher among Inuit men. Triglyceride was lower among Inuit of both sexes. Cholesterol and triglyceride varied according to westernisation, diet, alcohol consumption and smoking. In a multivariate analysis, serum lipids also differed significantly between pure and genetically mixed Inuit: HDL cholesterol was higher among the genetically pure Inuit, while among men triglyceride was lower and among women total and LDL-cholesterol were higher. INTERPRETATION: Among the Inuit, serum lipids are significantly associated with westernisation and genetic heritage. The effect of westernisation is to some extent due to dietary changes. From a cardiovascular health point of view, westernisation within Greenland is associated with unfavourable lipid changes while migration to Denmark is associated with favourable lipid changes. PMID- 15136073 TI - Monitoring plasma concentrations to individualize treatment with clomiphene citrate. AB - OBJECTIVE: Recent reports have indicated that a number of individual patient characteristics are responsible for the success or failure of clomiphene citrate treatment. However, a priori individualization of doses in different patients has not been investigated. We examined the thesis that wide variability in the metabolism of the active component (zuclomiphene) contributes to variability in response. METHODS: The dose-response relationship of clomiphene was established from a meta-analysis of data from 13 published reports. Limited data relating plasma drug concentrations to treatment outcome were examined to determine whether insufficient systemic exposure at a fixed dosage might contribute to therapeutic failure. RESULTS: A fixed-dosage regimen of 50 mg clomiphene per day is likely to cause ovulation in only 46% of patients; subsequent increment in dosage increases the number of responders but at the expense of considerable delay in individualization of treatment. Case reports indicated that dosage based on plasma drug concentration monitoring could improve patient management, and an algorithm is proposed to facilitate treatment. CONCLUSIONS: Prospective studies of clomiphene citrate should be performed to confirm the hypothesis that the monitoring of plasma zuclomiphene concentrations can significantly accelerate dose individualization and improve the therapeutic outcome with this "orphan" drug. PMID- 15136074 TI - Removal of endometriomas before in vitro fertilization does not improve fertility outcomes: a matched, case-control study. AB - OBJECTIVE: To investigate whether conservative surgery on ovarian endometriomas before an IVF cycle improves fertility outcomes. DESIGN: Retrospective, matched case-control study. SETTING: Two academic IVF programs. PATIENT(S): One hundred eighty-nine women with endometriomas who underwent IVF treatment: 56 women proceeded directly to IVF, and 133 first underwent conservative ovarian surgery. INTERVENTION(S): Controlled ovarian hyperstimulation and IVF-ET. MAIN OUTCOME MEASURE(S): Response to gonadotropins, fertilization, implantation, and pregnancy rates. RESULT(S): Aside from lower peak E(2) levels on the day of hCG and a higher total FSH dose in women previously operated for an endometrioma, no significant differences were found between the two groups in the different IVF variables analyzed. CONCLUSION(S): Laparoscopic cystectomy for endometriomas before commencing an IVF cycle does not improve fertility outcomes. Proceeding directly to controlled ovarian hyperstimulation in women with asymptomatic ovarian endometriomas might reduce the time to pregnancy, the costs of treatment, and the hypothetical complications of laparoscopic surgery. Conversely, conservative surgical treatment of ovarian endometriomas in symptomatic women does not impair IVF or intracytoplasmic sperm injection success rates. PMID- 15136075 TI - Endometriosis and the outcome of in vitro fertilization. AB - A generalized reproductive dysfunction rather than endometriotic implants is likely to have an impact on the outcome of IVF. PMID- 15136076 TI - Understanding endometriosis is the key to successful therapeutic management. AB - Understanding endometriosis on a molecular, biochemical, and genetic level will lead to better methods for the diagnosis and therapeutic management of this enigmatic disease. PMID- 15136077 TI - Management of endometriosis in fertility patients. AB - The management of endometriosis in fertility patients remains empiric. Adverse effects on every reproductive component have been reported. How to counsel couples regarding treatment and the effects of endometriosis on fertility is still uncertain. PMID- 15136078 TI - Surgery for the removal of endometriomas before in vitro fertilization does not increase implantation and pregnancy rates. AB - Laparoscopic cystectomy of large endometriomas before an IVF cycle does not increase the implantation and pregnancy rate. PMID- 15136079 TI - Assisted reproductive technology in the United States: 2000 results generated from the American Society for Reproductive Medicine/Society for Assisted Reproductive Technology Registry. AB - OBJECTIVE: To summarize the procedures and outcomes of ART initiated in the United States in 2000. DESIGN: Data were collected electronically using the SART Clinical Outcome Reporting System software and submitted to the American Society for Reproductive Medicine/ Society for Assisted Reproductive Technology Registry. PARTICIPANT(S): Three hundred eighty-three programs submitted data on procedures performed in 2000. Data were collated after November 2000 so that the outcome of all pregnancies established would be known. MAIN OUTCOME MEASURE(S): Incidence of clinical pregnancy, ectopic pregnancy, abortion, stillbirth, and delivery. RESULT(S): Programs reported initiating 99,989 cycles of ART treatment. Of these, 73,406 cycles involved fresh nondonor IVF (46.6% with intracytoplasmic sperm injection [ICSI]), with a delivery rate per retrieval of 29.9%; 549 were cycles of gamete intrafallopian transfer, with a delivery rate per retrieval of 24.7%; 763 were cycles of zygote intrafallopian transfer, with a delivery rate per retrieval of 29.9%. The following additional ART procedures were also initiated: 7,581 fresh donor oocyte cycles, with a delivery rate per transfer of 43.7%; 13,083 frozen embryo transfer procedures, with a delivery rate per transfer of 20.4%; 2,721 frozen embryo transfers using donated oocytes or embryos, with a delivery rate per transfer of 23.5%, and 1,200 cycles using a host uterus, with a delivery rate per transfer of 35.8%. In addition, 326 cycles were reported as combinations of more than one treatment type, 41 cycles as research, and 319 as embryo banking. As a result of all procedures, 25,394 deliveries were reported, resulting in 35,345 neonates, of which 35,031 were live born and 314 stillborn. CONCLUSION(S): In 2000, there were more programs reporting ART treatment and a significant (13.5%) increase in reported cycles compared to 1999. In comparable cycle types, overall success rate (deliveries per retrieval) exhibited an actual increase of 0.6%, which represents an increase of 2.2% when compared to the success rate for 1999. PMID- 15136080 TI - Previous ectopic pregnancy as a predictor of failure of systemic methotrexate therapy. AB - OBJECTIVE: To investigate whether a history of previous ectopic pregnancy is an independent risk factor for failure of methotrexate therapy in subsequent ectopic pregnancies and whether the previous treatment method influenced failure. SETTING: Inner-city teaching hospital. DESIGN: Retrospective case review. PATIENT(S): Five hundred four consecutive patients treated with "single-dose" methotrexate for ectopic pregnancy. Nine patients electing surgery after beginning medical therapy were excluded from analysis. INTERVENTION(S): Systemic methotrexate 50 mg/m(2). MAIN OUTCOME MEASURE(S): Failure of methotrexate therapy. RESULT(S): Twenty-one (18.6%) of 113 of previous ectopic patients and 26 (6.8%) of 382 first-time ectopic patients failed methotrexate therapy (odds ratio, 3.12). The overall success rate was 90.5% (448/495 patients). Univariant analysis showed that treatment hCG and progesterone (P) levels and ectopic cardiac activity were significantly different between successful and failed therapy. Logistic regression analysis for these four variables showed that only hCG and history of previous methotrexate remained significant risk factors for failure of methotrexate therapy. The likelihood of failure in patients with a previous ectopic was not influenced by previous treatment with salpingostomy, salpingectomy, or medical treatment. CONCLUSION(S): A history of previous ectopic pregnancy appears to be an independent risk factor for failure of systemic methotrexate treatment, but failure is not affected by previous treatment method. PMID- 15136081 TI - Ectopic pregnancy diagnosis and the pseudo-sac. AB - OBJECTIVE: To evaluate the impact of an ultrasound finding of a pseudo-sac (PS), a uterine sac without a double decidual ring or a yolk sac, on the management of cases with possible ectopic pregnancy. DESIGN: A retrospective review of a series of cases. SETTING: A general hospital. PATIENT(S): Seventy-seven patients who had a diagnostic laparoscopy over a period of 3 years for suspected ectopic pregnancy. MAIN OUTCOME MEASURE(S): A logistic regression analysis was conducted to evaluate the effect of the ultrasound finding of a PS on predicting a negative finding at laparoscopy. RESULT(S): The report of a PS is significantly associated with a false-positive diagnosis of ectopic pregnancy. CONCLUSION(S): A diagnosis of PS should not be interpreted as indicative of an ectopic pregnancy because radiological differentiation between an early intrauterine pregnancy failure and an ectopic pregnancy is not possible. PMID- 15136082 TI - Decreased levels of interleukin-18 in peritoneal fluid but not in serum of patients with endometriosis. AB - OBJECTIVE: To determine the concentrations of interleukin-18 (IL-18) in peritoneal fluid and serum in patients with endometriosis in comparison with the control group. DESIGN: A prospective analytical study. SETTING: The obstetrics and gynecology department of an academic training hospital. PATIENT(S): Forty four patients who underwent laparoscopic surgery for benign gynecologic diseases. INTERVENTION(S): Specimens of peripheral blood and peritoneal fluid were obtained before and during laparoscopic procedures, and the levels of IL-18 were analyzed. MAIN OUTCOME MEASURE(S): The concentrations of IL-18 in peritoneal fluid and serum were correlated with the presence of endometriosis, disease stage, and the phase of the menstrual cycle. RESULT(S): Interleukin-18 was detectable in 98% of the peritoneal specimens and 84% of the serum specimens of the patients tested. Peritoneal fluid IL-18 concentrations were statistically significantly lower in patients with endometriosis than in patients without endometriosis; the difference in serum IL-18 levels showed no statistically significant difference between the patients with and without endometriosis. The concentrations of IL-18 in peritoneal fluid and serum were not correlated with the stage of endometriosis or the phase of the menstrual cycle. CONCLUSION(S): Our results suggest that the decreased levels of IL-18 in peritoneal fluid in patients with endometriosis as compared with the control group may play an important role in the pathogenesis of this disease. PMID- 15136083 TI - Matrix metalloproteinase 2 is associated with changes in steroid hormones in the sera and peritoneal fluid of patients with endometriosis. AB - OBJECTIVE: To determine matrix metalloproteinase 2 (MMP-2) levels in sera and peritoneal fluids of women with or without endometriosis-associated infertility and to determine whether MMP-2 had correlation with 17beta-E(2) and P in the sera and peritoneal fluids of the patients with endometriosis. DESIGN: A case-control study. SETTING: A women's hospital in China. PATIENT(S): Forty patients with endometriosis-associated infertility undergoing laparoscopy and 18 women undergoing tubal ligation. INTERVENTION(S): Sera and peritoneal fluids were collected from women with or without endometriosis. MAIN OUTCOME MEASURE(S): Matrix metalloproteinase 2, 17beta-E(2), and P concentration. RESULT(S): Compared with the control women, the patients with endometriosis had higher MMP-2 levels in their sera and peritoneal fluids. Serum and peritoneal fluid MMP-2 levels were higher in the proliferative phase than in the secretory phase. The level of MMP-2 in the patients was correlated positively with 17beta-E(2) in serum and peritoneal fluid and negatively with P. CONCLUSION(S): The results demonstrated a correlation between MMP-2 concentrations and steroid hormones in sera and peritoneal fluids of the patients with endometriosis-associated infertility. Estradiol might up-regulate MMP-2, resulting in the formation of endometriosis. Progesterone might down-regulate MMP-2 to inhibit the formation and development of endometriosis. PMID- 15136084 TI - Cost analysis of singleton versus twin pregnancies after in vitro fertilization. AB - OBJECTIVE: To determine the difference in costs between singleton and twin pregnancies after IVF treatment from pregnancy to 6 weeks after delivery from a health care perspective. DESIGN: Retrospective cost analysis. SETTING: IVF department at the University Medical Center Nijmegen, The Netherlands. PATIENT(S): A representative sample of singleton and twin pregnancies after IVF treatment between 1995 and 2001 at the University Medical Center Nijmegen. INTERVENTION(S): IVF with or without intracytoplasmic sperm injection and with or without cryopreservation. MAIN OUTCOME MEASURE(S): Medical costs per singleton and twin pregnancy after IVF. RESULT(S): In patients pregnant with twins, the incidence of hospital antenatal care, complicated vaginal deliveries, and cesarean sections was higher and was associated with more frequent and longer maternal and neonatal hospital admissions. Maternal and neonatal hospital admissions were the major cost drivers. The medical cost per twin pregnancy was found to be more than five times higher than per singleton pregnancy, 13,469 and 2,550, respectively. CONCLUSION(S): The medical cost per twin pregnancy was more than 10,000 higher than per singleton pregnancy. A reduction in the number of twin pregnancies by elective single ET will save substantial amounts of money. This money might be used for the additional IVF cycles that will probably be needed to achieve similar success rates between single ET and two-embryo transfer. PMID- 15136085 TI - A poor response in the first in vitro fertilization cycle is not necessarily related to a poor prognosis in subsequent cycles. AB - OBJECTIVE: To calculate the cumulative ongoing pregnancy rate in patients with a poor response in their first IVF cycle. DESIGN: Retrospective cohort study. SETTING: In vitro fertilization unit of a university hospital. PATIENT(S): Two hundred twenty-five women who experienced a poor response in their first IVF or intracytoplasmic sperm injection cycle. These patients were divided into 64 expected (aged > or =41 years and/or elevated FSH level) and 161 unexpected poor responders (aged <41 years and FSH level not elevated). INTERVENTION(S): In vitro fertilization treatment with a long-suppression protocol with FSH-urofollitropin or recombinant FSH. MAIN OUTCOME MEASURE(S): Cumulative ongoing pregnancy rate. This rate was calculated in two ways to correct for dropouts: pessimistic (zero chance of pregnancy for the dropouts) and optimistic (the same chance for the dropouts as for patients who continued). RESULT(S): The cumulative ongoing pregnancy rate of women with an unexpected poor response in the first cycle was 37% (pessimistic) to 47% (optimistic) after three cycles. Women with an expected poor response had a cumulative ongoing pregnancy rate of 16% (pessimistic) to 19% (optimistic) after 3 cycles. Sixty-four percent of the unexpected poor responders and 31% of the expected poor responders had a normal response in the second cycle, most of them after receiving a higher dose of gonadotropins. CONCLUSION(S): Most patients with an unexpected poor response in the first cycle had a normal response in the second cycle, leading to an acceptable cumulative ongoing pregnancy rate after three cycles. Patients with an expected poor response in the first cycle should be advised to withdraw from treatment after the first cycle because of a poor prognosis. PMID- 15136086 TI - Emotional adaptation following successful in vitro fertilization. AB - OBJECTIVE: To assess the emotional impact of infertility after successful IVF and to compare parents who have undergone IVF (IVF parents) and parents who have not undergone IVF (non-IVF parents) regarding parental stress and the marital relationship during the transition to parenthood. DESIGN: A study with qualitative and longitudinal quantitative assessments. SETTING: University IVF clinics and antenatal clinics in Stockholm. PATIENT(S): Fifty-five IVF mothers, 53 IVF fathers, 40 non-IVF mothers, and 36 non-IVF fathers. INTERVENTION(S): IVF parents were interviewed. All subjects completed self-rating scales in early pregnancy and at 2 and 6 months postpartum. MAIN OUTCOME MEASURE(S): Interviews about perception of infertility and scalar measurement of parental stress and the marital relationship. RESULT(S): Negative feelings related to infertility were not easily overcome among the IVF parents. Their levels of stress related to parenthood were similar to those of non-IVF parents, and both groups reported decreased satisfaction with the marital relationship during the transition to parenthood. CONCLUSION(S): The inability to conceive naturally continues to affect the current lives of a proportion of IVF parents. The results suggest that IVF parents may benefit from counseling with regard to the potential long-term impacts of infertility, disclosure issues, and decisions regarding future children. However, levels of parental stress and patterns of partner satisfaction are similar to those of parents with children conceived "naturally." PMID- 15136087 TI - Relationship between maternal age and aneuploidy in in vitro fertilization pregnancy loss. AB - OBJECTIVE: To determine the fetal loss rate after documented fetal cardiac activity (7-week sonogram) and to evaluate the chromosomal makeup of these losses in IVF pregnancies. DESIGN: Retrospective analysis. SETTING: University-based IVF center. PATIENT(S): Two thousand fourteen consecutive IVF pregnancies with documented fetal cardiac activity. MAIN OUTCOME MEASURE(S): Miscarriage rates and karyotypes of pregnancy losses were analyzed. RESULT(S): The overall pregnancy loss rate after demonstrated fetal cardiac activity was 11.6% (233/2014). A highly significant increase in fetal loss with advancing maternal age was observed (<30 years = 5.3% vs. 31-34 years = 7.6% vs. 35-39 years = 12.8% vs. > or =40 years = 22.2%). Patients with a multiple gestation were more likely to deliver a live infant, compared with those with a singleton detected at a 7-week sonogram. Of the 233 losses in the study period, cytogenetic analyses were obtained for 74 (31.8%). Three specimens were nondiagnostic. Fifty-two patients had abnormal karyotypes (71.2% [52/71]). Eighty-two percent of the pregnancy losses in women aged > or =40 years were associated with chromosomally abnormal fetuses, compared with 65% of the losses in women aged <40 years (odds ratio, 3.35; 95% confidence interval, 0.96-11.97). CONCLUSION(S): Pregnancy loss after documentation of fetal cardiac activity is >10%. This loss is significantly increased with advancing maternal age. The major underlying cause of these losses seems to be chromosomal aneuploidy. PMID- 15136088 TI - Rate of aneuploidy in miscarriages following in vitro fertilization and intracytoplasmic sperm injection. AB - OBJECTIVE: To evaluate the incidence of aneuploidy in miscarriages after IVF and intracytoplasmic sperm injection (ICSI) procedures. DESIGN: Retrospective study. SETTING: University IVF program. PATIENT(S): All IVF patients with missed abortions undergoing uterine curettage. INTERVENTION(S): Cytogenetic analysis of products of conception (POC). MAIN OUTCOME MEASURE(S): Incidence of aneuploidy in POC. RESULT(S): Thirty-two of 59 specimens (54%) reviewed were abnormal. The patients with ICSI were more likely to have aneuploidy identified in their POC than conventional IVF, 76% vs. 41%. The average ages in these groups were similar: 37.1 vs. 37.8 years. There was a trend toward decreased aneuploidy with day 5 compared to day 3 embryo transfers; 38% vs. 63%. CONCLUSION(S): We found a significantly higher aneuploidy rate in the abortuses of patients who conceived with ICSI. It is possible that this increased incidence is due to abnormalities in the sperm of patients with ICSI, but could also be partially related to the technique itself. PMID- 15136089 TI - Middle to lower uterine segment embryo transfer improves implantation and pregnancy rates compared with fundal embryo transfer. AB - OBJECTIVE: To assess differences in pregnancy and implantation rates as a function of the embryo placement. DESIGN: Prospective cohort study. SETTING: A tertiary care center. SUBJECT(S): All fresh, nondonor IVF cycles performed in 2001. INTERVENTION(S): Alteration in embryo transfer (ET) target location from the fundal region to the middle to lower uterine segment. MAIN OUTCOME MEASURE(S): Clinical pregnancy rate (sonographic sac evidence/number of transfer cycles), implantation rate (number of sacs/number of embryos transferred), patient age, peak E(2), and fertilization rate. RESULT(S): A total of 393 fundal and 273 lower to middle uterine segment ETs were performed. The pregnancy (PR), implantation, and birth rates were significantly higher after a middle to lower uterine segment ET compared with fundal ET (39.6% vs. 31.2%; 21% vs. 14%; and 34.1% vs. 26.2%, respectively). Groups did not differ regarding patient age, basal FSH, peak E(2), number of intracytoplasmic sperm injection (ICSI) cycles, fertilization rate, embryo quality, or number of embryos transferred. CONCLUSION(S): Both PR and implantation rates are favorably affected by directing embryo placement to the lower to middle uterine segment. By some unknown mechanism, it appears that this endometrial location provides a more favorable region for embryo deposition. PMID- 15136090 TI - Effect of supraphysiologic doses of testosterone on fasting plasma total homocysteine concentrations in men with Klinefelter's syndrome. AB - OBJECTIVE: To examine the effect of supraphysiologic doses of testosterone (T) on plasma total homocysteine (tHcy) concentrations in patients with Klinefelter's syndrome (KS). DESIGN: Prospective clinical study. SETTING: University hospital. PATIENT(S): Thirty-two newly diagnosed male patients with KS, and 20 healthy, volunteer controls matched by age and body mass index. INTERVENTION(S): Testosterone was administered IM every 2 weeks for 6 months. Initially, pretreatment fasting blood samples were collected after overnight fasting. Posttreatment blood samples were drawn 7 days after the last injection. MAIN OUTCOME MEASURE(S): Plasma total homocysteine. RESULT(S): The KS patients had lower tHcy levels than the controls. However, plasma fasting tHcy concentrations increased in a statistically significant manner after 6 months of treatment. As compared with the levels among controls, pretreatment levels of the serum creatinine, hemoglobin, and hematocrit were significantly lower, and increased in a statistically significant way following treatment. Posttreatment levels of total cholesterol were statistically significantly higher than the baseline. The pretreatment folate and cobalamin levels also were statistically significantly higher in patients when compared with controls, and decreased significantly after treatment. The linear regression analysis showed that only creatinine, cobalamin, and folate were independently associated with plasma tHcy levels in patients before and after treatment. CONCLUSION(S): The patients with KS showed lower tHcy concentrations than healthy, age-matched male controls. Testosterone treatment increased plasma tHcy levels. PMID- 15136091 TI - Cytogenetic abnormalities and the failure of development after round spermatid injections. AB - OBJECTIVE: To assess cytologic and cytogenetic abnormalities following round spermatid injection. DESIGN: Prospective analysis. SETTING: In vitro fertilization centers. PATIENT(S): Fourteen couples accepted to a round spermatid injection (ROSI) and preimplantation genetic diagnosis (PGD) program after appropriate counseling. INTERVENTION(S): ROSI, PGD, with fluorescence in situ hybridization for chromosome enumeration. MAIN OUTCOME MEASURE(S): Cytologic and cytogenetic abnormalities in oocytes, zygotes, and blastomeres. RESULT(S): The fertilization rate following ROSI was 36%. Only 11 of 143 (7.7%) oocytes developed to have several blastomeres. Cytologic and cytogenetic abnormalities accounted for the vast majority of blockage at oocyte, zygote, and early mitotic division stages. Four biopsied embryos were normal. These and seven others were implanted, but no pregnancy was achieved. CONCLUSION(S): A PGD diagnosis for common aneuploidies and blastocyst stage transfer is feasible for ROSI cases. Failure with ROSI is cause primarily by chromosome abnormalities, so use of ROSI in assisted reproductive technologies should be limited. PMID- 15136092 TI - Sperm chromatin structure assay (SCSA) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles. AB - OBJECTIVE: To determine the relationship between sperm chromatin structure assay (SCSA) parameters (DNA fragmentation index [DFI] and high DNA stainability [HDS]), and conventional IVF and IVF/intracytoplasmic sperm injection (ICSI) outcomes. DESIGN: Retrospective review and prospective study. SETTING: Private IVF clinic. PATIENT(S): Two hundred forty-nine couples undergoing first IVF and/or ICSI cycle. INTERVENTION(S): IVF, ICSI, blastocyst culture. MAIN OUTCOME MEASURE(S): DFI, HDS, conventional semen parameters, IVF, ICSI. RESULT(S): IVF and ICSI fertilization rates were not statistically different between high- and low-DFI groups. More men with > or =15% HDS had lower (<25% and <50%) IVF fertilization rates. High DNA stainability was not related to ICSI fertilization rates. High DNA stainability did not affect blastocyst rates or pregnancy outcomes. Men with > or =30% DFI were at risk for low blastocyst rates (<30%) and no ongoing pregnancies. Men with > or =30% DFI had more male factors. World Health Organization thresholds were not predictive of ongoing pregnancy. CONCLUSION(S): The relationship between HDS and poor IVF fertilization rates provides preliminary evidence that ICSI may be indicated in men with > or =15% HDS. Men with high levels of DNA fragmentation (> or =30% DFI) were at greater risk for low blastocyst rates and failure to initiate an ongoing pregnancy. The SCSA provides valuable prognostic information to physicians counseling couples before IVF and/or ICSI cycles. PMID- 15136093 TI - Parental karyotype and subsequent live births in recurrent miscarriage. AB - OBJECTIVE: To compare the subsequent live birth rate in recurrently miscarrying women with and without parental balanced chromosomal aberrations. DESIGN: Retrospective comparative cohort study. SETTING: Tertiary referral unit in a university hospital. PATIENT(S): Nine hundred sixteen patients with 3-16 miscarriages before 20 weeks: 99 patients with and 817 patients without chromosomal aberrations. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Outcome of the subsequent pregnancy in terms of live births or repeat miscarriage. RESULT(S): Of the 916 patients, 661 subsequently conceived, 73 (73.7%) with parental chromosomal aberrations and 588 (71.9%) without aberrations. In patients with and without chromosomal aberrations, 33 of 73 pregnancies (45.2%) and 325 of 588 pregnancies (55.3%), respectively, resulted in live births. The difference is not statistically significant. There was a similar prevalence of aberrations in primary, secondary, and tertiary aborters. The prevalence of aberrations was not related to the number of previous miscarriages. Translocations, inversions, and mosaicism were followed by a similar live birth rate. CONCLUSION(S): Patients with parental chromosomal rearrangements do not have a significantly lower live birth rate than patients without aberrations. Parental karyotyping might not be a good predictor of the outcome of subsequent pregnancies. PMID- 15136094 TI - Preimplantation genetic diagnosis for the treatment of failed in vitro fertilization-embryo transfer and habitual abortion. AB - OBJECTIVE: To apply preimplantation genetic diagnosis (PGD) for the treatment of patients with a history of failed IVF-ET or habitual aborters. DESIGN: Prospective clinical study. SETTING: Tertiary center for assisted reproduction. PATIENT(S): Ninety-four couples with failed IVF-ET after >2 IVF cycles and 64 couples with >2 spontaneous abortions. INTERVENTION(S): Patients were prepared for oocyte retrieval using standard controlled ovarian hyperstimulation protocols after standard laboratory techniques. Blastomeres from 6- to 8-cell embryos were analysed using fluorescence in situ hybridization with commercial chromosomal probes, and normoploid embryos were transferred on day 3 after fertilization. MAIN OUTCOME MEASURE(S): Pregnancy and implantation rates and live births. RESULT(S): Both 3- and 5-probe PGD resulted in a significantly higher outcome than controls for failed IVF-ET. Five-probe PGD appeared to be more suitable for habitual aborters. CONCLUSION(S): This pilot study suggests that 3-probe PGD is a valid option for failed IVF-ET patients. The use of five or more probes is indicated for habitual aborters. PMID- 15136095 TI - Analysis of factors influencing pregnancy rates in homologous intrauterine insemination. AB - OBJECTIVE: To identify predictors of pregnancy rate (PR) among women undergoing homologous IUI. DESIGN: Cross-sectional analysis of IUI cycles carried out from January 2000 to September 2002. SETTING: Private infertility center in Alicante, Spain. PATIENT(S): Four hundred seventy women undergoing 1,010 cycles of IUI. INTERVENTION(S): Single IUI with ovarian stimulation using hMG. MAIN OUTCOME MEASURE(S): Preovulatory follicles (>15 mm), motile spermatozoa count, type and duration of infertility, female age, insemination timing, and cycle number. RESULT(S): Overall PR per cycle and multiple pregnancy and miscarriage rates were 9.2%, 8.6%, and 11.8%, respectively. Three significant predictors of pregnancy were identified by multiple logistic regression analysis: preovulatory follicles, spermatozoa count, and infertility duration. Interuterine insemination with three follicles almost tripled the PR with respect to only one, odds ratio (OR) = 2.89 (95% confidence interval [CI], 1.54-5.41). Compared with insemination with a motile sperm count >30 x, 20.1-30, 10.1-20, 5.1-10, and < or =5 x10(6), insemination progressively decreased the PR, from 15.3% in the highest category to 3.6% in the lowest (OR lowest/highest = 0.20 [95% CI: 0.09-0.45]), with a statistically significant dose-response trend. Infertility duration > or =3 years was marginally associated with a lower PR, OR = 0.65 (95% CI, 0.40-1.04). Overall, female age was not a significant predictor of pregnancy, and although PR slightly decreased beyond two IUI cycles and when a single IUI was performed 36 40 hours after hCG administration, results were not statistically significant. CONCLUSION(S): Homologous IUI achieves the best results with two or three induced follicles, a high motile spermatozoa count, and infertility duration <3 years, irrespective of female age and fertility history. PMID- 15136096 TI - Pregnancies in patients with congenital adrenal hyperplasia with complete or almost complete impairment of 21-hydroxylase activity. AB - OBJECTIVE: To show that, with appropriate therapy, women with classic congenital adrenal hyperplasia (CAH) can become pregnant. DESIGN: Observational clinical study. SETTING: University hospital. PATIENT(S): Adult young women with CAH: three with the salt-wasting form and four patients with simple virilizing CAH due to severe homozygous or compound heterozygous mutations of the CYP21B gene (deletions, I172N in exon 4 and nt656A/C-->G in intron 2) who wished to become pregnant. INTERVENTION(S): After confirmation in the first patient of the beneficial effect of additional treatment with fludrocortisone in lowering 17alpha-hydroxyprogesterone (17-OHP) levels, five other patients were treated with hydrocortisone as three daily doses at 8-hour intervals and fludrocortisone 0.1-0.2 mg daily divided into two to three doses. One patient received glucocorticoid alone. MAIN OUTCOME MEASURE(S): Treatment was controlled on the basis of morning salivary 17-OHP estimates and plasma renin concentrations. RESULT(S): Nine pregnancies occurred in six women. The course of the pregnancies (except one spontaneous abortion) was normal without any other modification of therapy. Only the women treated with hydrocortisone alone did not become pregnant. CONCLUSION(S): When treated with a combination of glucocorticoids and mineralocorticoids, sexually active patients with the classic phenotype of CAH can become pregnant. PMID- 15136097 TI - Circulating angiogenic factors during periovulation and the luteal phase of normal menstrual cycles. AB - OBJECTIVE: To measure serial serum concentrations of the angiogenic factors vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and angiogenin (ANG) in the periovulatory and secretory phase of normal menstrual cycles in healthy women and to determine their peaks, which might reflect the stage of their critical angiogenic action. DESIGN: Prospective study. SETTING: University departments of obstetrics and gynecology. PARTICIPANT(S): Thirty-three healthy Swedish women with regular menstrual cycles. INTERVENTION(S): Serial blood samples were collected from each woman. Luteinizing hormone surge was identified by testing morning urine. MAIN OUTCOME MEASURE(S): Circulating levels of VEGF, bFGF, and ANG. RESULT(S): Circulating peak concentrations were determined for VEGF on day 0 and 9 after ovulation, for bFGF on day 1 before ovulation and day 9 after ovulation, and for ANG on day 3 after ovulation. CONCLUSION(S): Circulating VEGF increased in a stage-dependent cyclic fashion. Basic FGF peaked during the late proliferative and mid secretory phase. Circulating ANG showed increased expression around the early secretory phase of the cycle. PMID- 15136098 TI - Successful spontaneous pregnancies in women older than 45 years. AB - OBJECTIVE: To characterize the reproductive history of women who delivered an infant after a spontaneous pregnancy after the age of 45 years. DESIGN: Observational study. SETTING: University departments. PATIENT(S): Two hundred nine women who conceived spontaneously and had their most recent delivery after the age of 45 years. INTERVENTION(S): Review of all individual medical charts for a complete history of all pregnancies. MAIN OUTCOME MEASURE(S): Gravidity, parity, and spontaneous abortion (SAB) rate. RESULT(S): The mean (+/-SD) age at last delivery was 45.7 +/- 0.9 years (range: 45-49). The mean parity was 9.6 +/- 4 children (range: 2-20): 81% of the women were grandmultiparas (> or =6 deliveries) and 46% were grand-grandmultiparas (> or =11 deliveries). The mean number of SAB was 1.9 +/- 1.9 (range: 0-9). Increased parity did not have a protective effect on abortion rate: 12.9% for parity 2-5, 15.6% for parity 6-10, and 21.3% for parity 11-20. Women in the study group differed significantly in their SAB rate from a general population: 11.3% vs. 17.7% at age 39, 13.2% vs. 33.8% at age 44, and 9.1% vs. 33.8% at age 45. CONCLUSION(S): Women who conceived at or after 45 years of age were mostly grandmultiparas. Although multiparity in itself did not affect SAB rates, this measure was remarkably low in our exceptionally fertile group as compared with the rates published for a general population. This unique group of grandmultiparous women might possess a genetic propensity that delays the normal rate of ovarian and oocyte senescence. PMID- 15136099 TI - A critical analysis of the accuracy, reproducibility, and clinical utility of histologic endometrial dating in fertile women. AB - OBJECTIVE: To refine or redefine the traditional histologic criteria used to date the secretory phase endometrium. DESIGN: Randomized, observational study. SETTING: Academic clinical research center. PATIENT(S): One hundred and thirty healthy, regularly cycling, fertile volunteers, aged 18 to 35 years. INTERVENTION(S): Patients were randomized to undergo endometrial sampling and measurement of serum estradiol and progesterone 1 to 14 days after the midcycle urinary luteinizing hormone surge. Three gynecologic histopathologists objectively scored each tissue specimen for 32 distinct histologic features and dated the endometrium using traditional histologic criteria. MAIN OUTCOME MEASURE(S): The 32 features were evaluated for [1] temporally dependent variation, [2] the amplitude of variations in score observed across the secretory phase, and [3] interobserver variability. Additionally, traditional dating criteria were analyzed. RESULT(S): The traditional endometrial histologic dating criteria are much less temporally distinct and discriminating than originally described, due to considerable intersubject, intrasubject, and interobserver variability. Neither traditional dating criteria nor any combination of the best performing histologic features identified by our objective and systematic analyses could reliably distinguish any specific cycle day or narrow interval of days. CONCLUSION(S): Histologic endometrial dating does not have the accuracy or the precision necessary to provide a valid method for the diagnosis of luteal phase deficiency or to otherwise guide the clinical management of women with reproductive failure. PMID- 15136100 TI - Preferences for fertility in women with pelvic inflammatory disease. AB - OBJECTIVE: To identify the value that women with pelvic inflammatory disease (PID) assign to the health impact of future infertility. DESIGN: Cross-sectional observations on patient preferences. SETTING: Participants in an existing multicenter clinical trial of PID treatment options. PATIENT(S): Five hundred thirty-two women with signs and symptoms of PID who were identified from emergency departments and sexually transmitted disease clinics. INTERVENTION(S): Women were asked to rate whether life with future infertility was more or less meaningful than life with each of seven chronic health conditions: sinus congestion, insomnia, chronic headache, asthma, incontinence, dialysis, and paralysis. MAIN OUTCOME MEASURE(S): Preferences regarding future infertility. RESULT(S): Most respondents rated future infertility as being worse than sinus congestion and asthma but better than the health impact of incontinence, dialysis, and paralysis. There was a wide range of opinion, with 18% viewing future infertility as minor (better than all conditions) and 5% viewing it as extremely important (worse than all conditions). Future infertility ratings were influenced by race, parity, difficulty in conceiving, and views on the importance of future pregnancy. CONCLUSION(S): The majority of women with PID rate future infertility as a significant issue. Optimizing access to infertility treatment may affect the quality of life for such women. PMID- 15136101 TI - Estrogen up-regulates cyclooxygenase-2 via estrogen receptor in human uterine microvascular endothelial cells. AB - OBJECTIVE: To investigate the effects of 17beta-estradiol (E(2)) on cyclooxygenase-2 (COX-2) expression and prostaglandin E(2) (PGE(2)) synthesis in primary human uterine microvascular endothelial cells (HUMEC). DESIGN: Prospective study. SETTING: Basic research laboratory at an academic medical center. PATIENT(S): Primary HUMEC of three women donors and primary human dermal microvascular endothelial cells of three women donors (as control), purchased from a third-party source. INTERVENTION(S): The HUMEC were cultured in specific media in a humidified atmosphere with 5% CO(2) at 37 degrees C. MAIN OUTCOME MEASURE(S): Measures of COX-2 mRNA and protein, PGE(2) production, and estrogen receptor alpha and beta mRNA and protein. RESULT(S): Treatment with E(2) (10(-10) to 10(-6) M) increased COX-2 mRNA levels by 2.3-fold to 2.4-fold in HUMEC. Treatment of HUMEC with E(2) (10(-8) M) resulted in a time-dependent increase of COX-2 mRNA levels. This was accompanied by a 2.8-fold increase in COX-2 protein level and a 1.5-fold increase in PGE(2) synthesis. Pretreatment of HUMEC with a selective COX-2 inhibitor, NS-398, abolished E(2)-induced PGE(2) synthesis, suggesting that E(2) specifically up-regulates COX-2 activity. The estrogen receptor antagonist ICI 182,780 fully reversed the stimulation of COX-2 mRNA and protein levels and PGE(2) synthesis by E(2). Interestingly, estrogen receptor beta mRNA and protein were abundant in HUMEC, whereas estrogen receptor alpha mRNA or protein was barely detectable. CONCLUSION(S): We conclude that various levels of E(2) can significantly increase COX-2 expression and PGE(2) synthesis in HUMEC via the estrogen receptor. PMID- 15136102 TI - Preclinical evaluation of magainin-A as a contraceptive antimicrobial agent. AB - OBJECTIVE: To evaluate the safety and contraceptive efficacy of magainin-A in monkeys. DESIGN: Controlled laboratory study. SETTING: Department of Immunology, National Institute for Research in Reproductive Health, Parel, Mumbai, India. ANIMAL(S): Male and female bonnet monkeys (Macaca radiata). INTERVENTION(S): Animals were treated intravaginally with 1 mg of magainin-A before attempted conception, as well as daily for 14 days to assess local and systemic toxicity. MAIN OUTCOME MEASURE(S): Suitability of magainin-A for the control of pregnancy and sexually transmitted infections. RESULT(S): Complete sperm immobilization was observed within 20 seconds after the exposure to magainin-A (800-1,000 microg) in vitro. Intravaginal administration of 1 mg of magainin-A blocked conception in monkeys. When magainin-A was administered intravaginally for 14 consecutive days, no treatment-related abnormalities were observed in menstrual cycle length, vaginal epithelial cell morphology, and hematologic/serum biochemical profiles. The peptide inhibited the growth of sexually transmitted infection-causing pathogens but not HIV-1 and HIV-2. CONCLUSION(S): Magainin-A can be used as an effective and safe intravaginal contraceptive compound with additional protection against sexually transmitted infection-causing pathogens. PMID- 15136103 TI - Air in the transfer catheter does not affect the success of embryo transfer. AB - OBJECTIVE: To investigate the effect on embryo transfer (ET) success of air loaded into the transfer catheter to bracket the embryo-containing medium. DESIGN: Prospective, randomized study. SETTING: University teaching hospital. PATIENT(S): One hundred two consecutive patients undergoing ET after IVF. INTERVENTION(S): In group 1 (n = 52), embryos were loaded as follows: 200 microL of air in the syringe, 100-125 microL of air in the proximal part of the catheter, 20-25 microL of medium containing the embryos to be transferred, and 10 microL of air at the tip of the catheter. In group 2 (n = 50), the syringe and the entire catheter were filled with medium and the embryo-containing medium (20 25 microL) was aspirated without being bracketed by air spaces. MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates. RESULT(S): No differences were found between groups 1 and 2 with respect to implantation and pregnancy rates. CONCLUSION(S): The air loaded into the transfer catheter to bracket the embryo containing medium has no negative effect on ET success. PMID- 15136104 TI - Superior outcomes of microsurgical vasectomy reversal in men with the same female partners. AB - OBJECTIVE: To evaluate the outcomes of vasectomy reversals in men with the same female partners. DESIGN: Retrospective analysis with comparison with a historical cohort. SETTING: University hospital. PATIENT(S): Among 1,048 patients who underwent microsurgical vasectomy reversal from 1986 to 2002, 27 men (2.6%) were identified who had the same partners as before their vasectomy. INTERVENTION(S): Microsurgical vasovasostomy or vasoepididymostomy. MAIN OUTCOME MEASURE(S): Semen parameters, clinical pregnancy, and live birth rates. RESULT(S): The mean age of the men was 38.5 years, and 37.2 years for their female partners. The reasons for vasectomy reversal were death of a child in 33% of cases and desire for more children in 66% of cases. The overall patency rate was 100% at 1 month postoperatively, with an average sperm concentration of 30 million/mL and 24% motility. Among patients with follow-up beyond 1 year, the natural pregnancy rate was 86%, which was achieved at 8.3 months postoperatively, with a live birth rate of 82%. The live birth rate in couples with the death of a child was 100%. CONCLUSION(S): The outcomes of vasectomy reversal in men with the same female partners are better than for men with new partners. Possible reasons for these superior results are previous proven fecundity as a couple, shorter time interval since vasectomy, and emotional dedication. PMID- 15136105 TI - Quantified short-term outcome of uterine artery embolization with gelatin sponge particles and lipiodol for symptomatic myoma. AB - OBJECTIVE: To investigate and quantify clinical outcomes and spectral Doppler analyses of uterine arteries in patients with myoma undergoing uterine artery embolization (UAE) with gelatin sponge particles and lipiodol. DESIGN: Prospective observational study. SETTING: Tertiary medical center. PATIENT(S): Forty premenopausal women with symptomatic myoma. INTERVENTION(S): Uterine artery embolization with gelatin sponge particles and lipiodol. MAIN OUTCOMES MEASURE(S): Hemoglobin, hematocrit, CA-125, pictorial blood loss assessment, visual analogue pain scale, questionnaire for symptoms, tumor volume, and spectral Doppler analyses of uterine arteries. RESULT(S): The mean follow-up period was 8.1 months (range, 6-12). Menstrual flow improved in 29 of 35 patients (83%) and decreased significantly by 78.4%. Menstrual pain improved in 27 of 35 patients (77%) and decreased significantly by 70%. Hematocrit and CA-125 improved significantly. The mean percentage reductions of uterine and myomal volumes were 40.2% and 54.9%, respectively. The mean peak systolic velocity of the uterine arteries decreased by 52%. The major complication rate was 2.56%. There was no correlation between tumor volume reduction and clinical outcome. CONCLUSION(S): Uterine artery embolization with gelatin sponge particles and lipiodol had satisfactory short-term outcomes, comparable to those associated with polyvinyl alcohol particles. Quantified and semiquantified measurements provided objective assessment of clinical outcomes. Serum CA-125 might play a role in clinical follow-up. Reduction of tumor volume is not predictive of UAE efficacy. PMID- 15136106 TI - Lesson learned and dispelled myths: three-dimensional imaging of the human vagina. AB - Three-dimensional imaging of the human vagina demonstrates that the cross section can be a "W," rather than an "H," and that intravaginal gel can ascend into the endocervix and presumably into the endometrium. PMID- 15136107 TI - Development of leiomyomas on the uterine remnants of two women with Mayer Rokitansky-Kuster-Hauser syndrome. AB - OBJECTIVE: To describe two cases of leiomyoma development in patients with Mayer Rokitansky-Kuster-Hauser syndrome (MRKH syndrome). DESIGN: Case report. SETTING: Second Department of Obstetrics and Gynecology, University of Athens, Aretaieion Hospital, Athens, Greece. PATIENT(S): A 42-year-old woman with MRKH syndrome presented with lower abdominal pain, and a 38-year-old woman with MRKH syndrome presented with an asymptomatic left adnexal mass. INTERVENTIONS(S): Clinical examination, transabdominal ultrasonography, IV urography, laparoscopy. MAIN OUTCOME MEASURE(S): Ultrasound. RESULT(S): In both cases, laparoscopy revealed a leiomyoma originating from the left uterine remnant. The leiomyomas and the adjacent uterine remnants were laparoscopically excised. CONCLUSION(S): In rare cases, leiomyomas can originate from the fibromuscular tissue of uterine remnants in patients with MRKH syndrome. PMID- 15136108 TI - Molecular and cytogenetic characterization of a structural rearrangement of the Y chromosome in an azoospermic man. AB - OBJECTIVE: To better define an abnormal karyotype found in a male with primary infertility. DESIGN: Case report. SETTING: Molecular and cytogenetics unit in a university-affiliated hospital. PATIENT(S): A 41-year-old, azoospermic, but otherwise healthy male. INTERVENTION(S): Lymphocytic karyotype and genetic counseling. MAIN OUTCOME MEASURE(S): Metaphases were studied by standard G- and Q banding, followed by fluorescence in situ hybridization (FISH) and polymerase chain reaction to analyze specific Y chromosome regions. RESULT(S): Chromosomal analysis and FISH allowed us to define the propositus's karyotype as 45,X/46,X,idic(Yp)/46,XY (71%, 26%, and 3% of analyzed metaphases, respectively). Molecular analysis of azoospermic factor (AZF) regions showed deletion of AZFb and AZFc. CONCLUSION(S): A 45,X/46,X,idic(Yp) mosaicism is associated with a very broad spectrum of phenotypes, including patients with Ullrich-Turner syndrome, patients with various degrees of genital ambiguity, or normal males. In the presence of a normal masculinization in otherwise healthy males azoospermia is a distinct feature that can be explained by partial deletion of AZF regions. PMID- 15136109 TI - Sertoli cell inactivation by cytotoxic damage to the human testis after cancer chemotherapy. AB - OBJECTIVE: To assess Sertoli cell involvement in postchemotherapy azoospermia. DESIGN: Case report. SETTING: Teaching hospital. PATIENT(S): A 31-year-old azoospermic man who underwent cancer cytotoxic chemotherapy for non-Hodgkin's lymphoma at 13 years of age. INTERVENTION(S): Testicular biopsy specimens were obtained for sperm recovery in preparation for intracytoplasmic sperm injection. The biopsy specimens were evaluated by quantitative immunohistochemistry for the immature Sertoli cell markers cytokeratin 18 (CK-18) and D2-40. MAIN OUTCOME MEASURE(S): Extent of immature Sertoli cells. RESULT(S): A fraction of Sertoli cells (13%) in the atrophic tubules of this patient reexpressed the intermediate filament protein CK-18, which is normally absent after puberty, but not the D2-40 antigen, an Mr 40,000 a-linked membrane glycoprotein, whose loss of expression at puberty marks an irreversible step in Sertoli cell maturation. Tubules with normal spermatogenic progression lined by Sertoli cells negative for CK-18 were also observed. CONCLUSION(S): A fraction of Sertoli cells of this patient initially progressed to full maturation at puberty and reverted to a dedifferentiated state marked by reexpression of CK-18 as a consequence of chemotherapy. This inactivation of Sertoli cells caused by the cytotoxicity of the chemotherapeutic drugs may have contributed to the spermatogenic impairment and resulting infertility. PMID- 15136110 TI - Successful treatment of severe endometriosis in two premenopausal women with an aromatase inhibitor. AB - OBJECTIVE: To treat severe endometriosis in premenopausal women with an aromatase inhibitor and to document the efficacy and side effects of this new treatment. DESIGN: Case report. SETTING: Clinical practice setting. PATIENT(S): Patients were premenopausal endometriosis cases confirmed by prior laparoscopy. INTERVENTION(S): Oral administration of the aromatase inhibitor anastrozole with the addition of 200-mg oral progesterone (P) capsules once daily, hs, and calcitriol 0.5 microg oral capsules twice daily. All three medications were given daily for 21 days followed by 7 days off, for a 28-day treatment cycle. Additionally, rofecoxib 12.5 mg was administered once a day continuously for the 28-day cycle. The dose could be increased to 25-50 mg daily as needed for increased pain. Six repeat 28-day cycles were planned for the treatment course depending on patient tolerance and response. MAIN OUTCOME MEASURE(S): Reduction of symptoms, elimination of endometrial implants, and improvement in fertility potential in the future. Monitoring for ovarian hyperstimulation as a side effect of aromatase inhibitor therapy. Documentation of potential symptoms or side effects when an aromatase inhibitor is used in menstruating women. RESULT(S): Treatment resulted in a rapid, progressive reduction in symptoms over 3 months with the maintenance of remission of symptoms for over 24 months after treatment in both cases. There was confirmation of absence of disease in one case by follow up laparoscopy 15 months after treatment. Pregnancy was achieved in both cases after 24 months. CONCLUSION(S): This is the first case report of successful treatment of severe endometriosis in premenopausal women with the aromatase inhibitor anastrozole combined with P, rofecoxib, and calcitriol. Treatment resulted in a rapid elimination of symptoms and was well tolerated in both cases. PMID- 15136111 TI - Effect of maximal endometrial thickness on outcome after frozen embryo transfer. AB - The largest study to date on the association of endometrial thickness and subsequent pregnancy rates following frozen embryo transfer with the endometrium prepared by estrogen and progesterone found no improved or adverse outcome if the endometrial thickness was > or = standard deviations above the mean. Neither was there a trend noted for thin endometria. PMID- 15136112 TI - Exogenous luteinizing hormone activity may influence the treatment outcome in in vitro fertilization but not in intracytoplasmic sperm injection cycles. AB - Additional analysis of a multinational, open-label, randomized study comparing highly purified hMG and recombinant FSH in a long protocol revealed that LH activity might favorably influence pregnancy outcome in IVF cycles. PMID- 15136113 TI - Impact of duration of cryopreservation of spermatozoa obtained through testicular sperm extraction on intracytoplasmic sperm injection. AB - Cryopreservation of testicular spermatozoa is feasible for patients suffering obstructive or nonobstructive azoospermia. A stndardized intracytoplasmic sperm injection procedure using frozen-thawed testicular tissue gives rise to fertilization and cleavage rates, which appear not to be affected by the duration of the period of cryostorage. PMID- 15136114 TI - Use of color Doppler sonography during follicular aspiration in patients undergoing in vitro fertilization may reduce the risk of blood vessel injury. AB - The use of color Doppler to identify small blood vessels (diameter: 0.68 +/- 0.17 mm to 2.28 +/- 0.81 mm; peak systolic velocity: 10.3 +/- 4.15 cm/s to 25.15 +/- 7.68 cm/s) during follicular aspiration may reduce patient's morbidity. PMID- 15136115 TI - Association of peroxisome proliferator-activated receptor gamma 2 Pro-12-Ala polymorphism with endometriosis. AB - We explored the association of the PPAR-gamma2 (peroxisome proliferator-activated receptor) Pro-12-Ala polymorphism with endometriosis in a case-control study with 51 women with endometriosis stages I-IV and 55 control women without endometriosis. The 12-Pro allele of PPAR-gamma2 may have protective effects avoiding the development and progression of endometriosis. PMID- 15136116 TI - "First uterine pass effect" is observed when estradiol is placed in the upper but not lower third of the vagina. AB - In postmenopausal women, the "first uterine pass effect" is seen when E(2) is placed in the upper but not lower third of the vagina. PMID- 15136117 TI - Overcoming maturation arrest by in vitro spermatogenesis: search for the optimal culture system. AB - In some men with nonobstructive azoospermia caused by germ cell maturation arrest, spermatogenesis can be reactivated in vitro, either by culturing segments of explanted seminiferous tubules or by coculturing isolated germ cells on monolayers of somatic cell lines. Further studies are needed to compare the efficacy of these two culture systems in terms of restoration of fertilizing ability of in vitro-formed gametes and to evaluate the safety of the clinical use of these cells in assisted reproduction treatment. PMID- 15136118 TI - Multifetal pregnancy reduction of triplet and higher-order multiple pregnancies to twins. AB - This systematic review was performed to assess the effects of multifetal pregnancy reduction for women with triplet and higher-order multiple pregnancies on fetal loss, preterm birth, and perinatal and infant mortality and morbidity. From nonrandomized studies, multifetal pregnancy reduction seems to be an effective treatment option, with outcomes comparable to those obtained from twin pregnancies conceived spontaneously or after assisted reproductive techniques. PMID- 15136119 TI - Clinically relevant differences and "Biocreep". PMID- 15136121 TI - Clinically relevant differences and "Biocreep". PMID- 15136122 TI - Ovarian reserve and a slightly different perspective. PMID- 15136124 TI - A role for "acquired" activated protein C resistance in recurrent fetal loss? PMID- 15136126 TI - Output and input from the controversy: recombinant FSH versus hMG. PMID- 15136128 TI - A genetic polymorphism and male fertility. PMID- 15136130 TI - Heparin/aspirin therapy for implantation failure in IVF cycles. PMID- 15136132 TI - Clues as to the molecular basis for uterine fibroids. PMID- 15136134 TI - Clues as to the molecular basis for uterine fibroids. PMID- 15136135 TI - Treatment of androgen deficiency in the aging male. AB - Although guidelines for androgen replacement therapy for older men have been developed by several groups, the data to support treatment are based on the results of only a small number of clinical trials. It has not been definitively established that the decline in testosterone seen in most aging men results in an androgen deficient state with health-related outcomes that can be improved by androgen therapy. PMID- 15136136 TI - Endometriosis and infertility. AB - Women with endometriosis typically present with pelvic pain, infertility or an adnexal mass. Surgery for persistent adnexal masses may be indicated to remove an endometrioma or other pelvic pathology. Surgical or medical therapy is efficacious for pelvic pain due to endometriosis, but treatment of endometriosis in the female partner of an infertile couple raises a number of complex clinical questions that do not have simple answers. PMID- 15136137 TI - Molecular and genetic regulation of testis descent and external genitalia development. AB - Testicular descent as a prerequisite for the production of mature spermatozoa and normal external genitalia morphogenesis, and therefore facilitating copulation and internal fertilization, are essential developmental steps in reproduction of vertebrate species. Cryptorchidism, the failure of testis descent, and feminization of external genitalia in the male, usually in the form of hypospadias, in which the opening of the urethra occurs along the ventral aspect of the penis, are the most frequent pediatric complications. Thus, elucidating the molecular mechanisms involved in the regulation of testis descent and the formation of external genitalia merits a special focus. Natural and transgenic rodent models have demonstrated both morphogenic processes to be under the control of a plethora of genetic factors with complex time-, space-, and dose restricted expression pattern. The review elucidates the molecular mechanisms involved in the regulation of testis descent and the formation of external genitalia and, wherever possible, assesses the differences between these rodent animal models and other mammalian species, including human. PMID- 15136138 TI - Follistatin complexes Myostatin and antagonises Myostatin-mediated inhibition of myogenesis. AB - Follistatin is known to antagonise the function of several members of the TGF beta family of secreted signalling factors, including Myostatin, the most powerful inhibitor of muscle growth characterised to date. In this study, we compare the expression of Myostatin and Follistatin during chick development and show that they are expressed in the vicinity or in overlapping domains to suggest possible interaction during muscle development. We performed yeast and mammalian two-hybrid studies and show that Myostatin and Follistatin interact directly. We further show that single modules of the Follistatin protein cannot associate with Myostatin suggesting that the entire protein is required for the interaction. We analysed the interaction kinetics of the two proteins and found that Follistatin binds Myostatin with a high affinity of 5.84 x 10(-10) M. We next tested whether Follistatin suppresses Myostatin activity during muscle development. We confirmed our previous observation that treatment of chick limb buds with Myostatin results in a severe decrease in the expression of two key myogenic regulatory genes Pax-3 and MyoD. However, in the presence of Follistatin, the Myostatin-mediated inhibition of Pax-3 and MyoD expression is blocked. We additionally show that Myostatin inhibits terminal differentiation of muscle cells in high-density cell cultures of limb mesenchyme (micromass) and that Follistatin rescues muscle differentiation in a concentration-dependent manner. In summary, our data suggest that Follistatin antagonises Myostatin by direct protein interaction, which prevents Myostatin from executing its inhibitory effect on muscle development. PMID- 15136139 TI - A Titin mutation defines roles for circulation in endothelial morphogenesis. AB - Morphogenesis of the developing vascular network requires coordinated regulation of an extensive array of endothelial cell behaviors. Precisely regulated signaling molecules such as vascular endothelial growth factor (VEGF) direct some of these endothelial behaviors. Newly forming blood vessels also become subjected to novel biomechanical forces upon initiation of cardiac contractions. We report here the identification of a recessive mouse mutation termed shrunken-head (shru) that disrupts function of the Titin gene. Titin was found to be required for the initiation of proper heart contractions as well as for maintaining the correct overall shape and orientation of individual cardiomyocytes. Cardiac dysfunction in shrunken-head mutant embryos provided an opportunity to study the effects of lack of blood circulation on the morphogenesis of endothelial cells. Without blood flow, differentiating endothelial cells display defects in their shapes and patterns of cell-cell contact. These endothelial cells, without exposure to blood circulation, have an abnormal distribution within vasculogenic vessels. Further effects of absent blood flow include abnormal spatial regulation of angiogenesis and elevated VEGF signaling. The shrunken-head mutation has provided an in vivo model to precisely define the roles of circulation on cellular and network aspects of vascular morphogenesis. PMID- 15136140 TI - BMP receptor IA is required in the mammalian embryo for endodermal morphogenesis and ectodermal patterning. AB - BMPRIA is a receptor for bone morphogenetic proteins with high affinity for BMP2 and BMP4. Mouse embryos lacking Bmpr1a fail to gastrulate, complicating studies on the requirements for BMP signaling in germ layer development. Recent work shows that BMP4 produced in extraembryonic tissues initiates gastrulation. Here we use a conditional allele of Bmpr1a to remove BMPRIA only in the epiblast, which gives rise to all embryonic tissues. Resulting embryos are mosaics composed primarily of cells homozygous null for Bmpr1a, interspersed with heterozygous cells. Although mesoderm and endoderm do not form in Bmpr1a null embryos, these tissues are present in the mosaics and are populated with mutant cells. Thus, BMPRIA signaling in the epiblast does not restrict cells to or from any of the germ layers. Cells lacking Bmpr1a also contribute to surface ectoderm; however, from the hindbrain forward, little surface ectoderm forms and the forebrain is enlarged and convoluted. Prechordal plate, early definitive endoderm, and anterior visceral endoderm appear to be expanded, likely due to defective morphogenesis. These data suggest that the enlarged forebrain is caused in part by increased exposure of the ectoderm to signaling sources that promote anterior neural fate. Our results reveal critical roles for BMP signaling in endodermal morphogenesis and ectodermal patterning. PMID- 15136141 TI - The Caenorhabditis elegans aryl hydrocarbon receptor, AHR-1, regulates neuronal development. AB - The mammalian aryl hydrocarbon receptor (AHR) is a ligand-activated transcription factor that mediates the toxic effects of dioxins and related compounds. Dioxins have been shown to cause a range of neurological defects, but the role of AHR during normal neuronal development is not known. Here we investigate the developmental functions of ahr-1, the Caenorhabditis elegans aryl hydrocarbon receptor homolog. We show that ahr-1:GFP is expressed in a subset of neurons, and we demonstrate that animals lacking ahr-1 function have specific defects in neuronal differentiation, as evidenced by changes in gene expression, aberrant cell migration, axon branching, or supernumerary neuronal processes. In ahr-1 deficient animals, the touch receptor neuron AVM and its sister cell, the interneuron SDQR, exhibit cell and axonal migration defects. We show that dorsal migration of SDQR is mediated by UNC-6/Netrin, SAX-3/Robo, and UNC-129/TGFbeta, and this process requires the functions of both ahr-1 and its transcription factor dimerization partner aha-1. We also document a role for ahr-1 during the differentiation of the neurons that contact the pseudocoelomic fluid. In ahr-1 deficient animals, these neurons are born but they do not express the cell-type specific markers gcy-32:GFP and npr-1:GFP at appropriate levels. Additionally, we show that ahr-1 expression is regulated by the UNC-86 transcription factor. We propose that the AHR-1 transcriptional complex acts in combination with other intrinsic and extracellular factors to direct the differentiation of distinct neuronal subtypes. These data, when considered with the neurotoxic effects of AHR activating pollutants, support the hypothesis that AHR has an evolutionarily conserved role in neuronal development. PMID- 15136142 TI - Runx2 mediates FGF signaling from epithelium to mesenchyme during tooth morphogenesis. AB - Runx2 (Cbfa1) is a runt domain transcription factor that is essential for bone development and tooth morphogenesis. Teeth form as ectodermal appendages and their development is regulated by interactions between the epithelium and mesenchyme. We have shown previously that Runx2 is expressed in the dental mesenchyme and regulated by FGF signals from the epithelium, and that tooth development arrests at late bud stage in Runx2 knockout mice [Development 126 (1999) 2911]. In the present study, we have continued to clarify the role of Runx2 in tooth development and searched for downstream targets of Runx2 by extensive in situ hybridization analysis. The expression of Fgf3 was downregulated in the mesenchyme of Runx2 mutant teeth. FGF-soaked beads failed to induce Fgf3 expression in Runx2 mutant dental mesenchyme whereas in wild-type mesenchyme they induced Fgf3 in all explants indicating a requirement of Runx2 for transduction of FGF signals. Fgf3 was absent also in cultured Runx2-/- calvarial cells and it was induced by overexpression of Runx2. Furthermore, Runx2 was downregulated in Msx1 mutant tooth germs, indicating that it functions in the dental mesenchyme between Msx1 and Fgf3. Shh expression was absent from the epithelial enamel knot in lower molars of Runx2 mutant and reduced in upper molars. However, other enamel knot marker genes were expressed normally in mutant upper molars, while reduced or missing in lower molars. These differences between mutant upper and lower molars may be explained by the substitution of Runx2 function by Runx3, another member of the runt gene family that was upregulated in upper but not lower molars of Runx2 mutants. Shh expression in mutant enamel knots was not rescued by FGFs in vitro, indicating that in addition to Fgf3, Runx2 regulates other mesenchymal genes required for early tooth morphogenesis. Also, exogenous FGF and SHH did not rescue the morphogenesis of Runx2 mutant molars. We conclude that Runx2 mediates the functions of epithelial FGF signals regulating Fgf3 expression in the dental mesenchyme and that Fgf3 may be a direct target gene of Runx2. PMID- 15136143 TI - EphA4, RhoB and the molecular development of feather buds are maintained by the integrity of the actin cytoskeleton. AB - The development of feather buds is a highly ordered process involving epithelial mesenchymal signalling. Cellular morphology is determined by the actin cytoskeleton, which is controlled by networks of regulators such as the GTPases. EphA4 belongs to a receptor tyrosine kinase family that has been consistently shown to regulate the cytoskeleton via Rho family GTPases in neural development and is expressed in early stages of feather bud development though its role has not been defined. We therefore used an in vitro skin culture system to interfere with EphA4 levels in feather buds using anti-sense oligonucleotides, demonstrating a severe effect on both their number and morphological form. Analysis of the Rho family of GTPases revealed that this effect was mediated by the GTPase RhoB, the expression of which was altered in response to altered levels of EphA4. In addition, the inhibition of RhoB mimicked the effects of reduced EphA4 levels on feather development. Significantly, manipulation of cytoskeletal dynamics revealed that those cells undergoing morphogenetic change regulate the patterning signals responsible for initiating feather development. We propose that this molecular maintenance mechanism between EphA4-RhoB and the actin cytoskeleton converges or coordinates with other morphogenic signalling systems to control feather bud development. PMID- 15136144 TI - The Drosophila RCC1 homolog, Bj1, regulates nucleocytoplasmic transport and neural differentiation during Drosophila development. AB - The Bj1 gene encodes the Drosophila homolog of RCC1, the guanine-nucleotide exchange factor for RanGTPase. Here, we provide the first phenotypic characterization of a RCC1 homolog in a developmental model system. We identified Bj1 (dRCC1) in a genetic screen to identify mutations that alter central nervous system development. We find that zygotic dRCC1 mutant embryos exhibit specific defects in the development and differentiation of lateral CNS neurons although cell division and the cell cycle appear grossly normal. dRCC1 mutant nerve cords contain abnormally large cells with compartmentalized nuclei and exhibit increased transcription in the lateral CNS. As RCC1 is an important component of the nucleocytoplasmic transport machinery, we find that dRCC1 function is required for nuclear import of nuclear localization signal sequence (NLS) carrying cargo molecules. Finally, we show that dRCC1 is required for cell proliferation and/or survival during germline, eye and wing development and that dRCC1 appears to facilitate apoptosis. PMID- 15136145 TI - The ladybird homeobox genes are essential for the specification of a subpopulation of neural cells. AB - In Drosophila, neurons and glial cells are produced by neural precursor cells called neuroblasts (NBs), which can be individually identified. Each NB generates a characteristic cell lineage specified by a precise spatiotemporal control of gene expression within the NB and its progeny. Here we show that the homeobox genes ladybird early and ladybird late are expressed in subsets of cells deriving from neuroblasts NB 5-3 and NB 5-6 and are essential for their correct development. Our analysis revealed that ladybird in Drosophila, like their vertebrate orthologous Lbx1 genes, play an important role in cell fate specification processes. Among those cells that express ladybird are NB 5-6 derived glial cells. In ladybird loss-of-function mutants, the NB 5-6-derived exit glial cells are absent while overexpression of these genes leads to supernumerary glial cells of this type. Furthermore, aberrant glial cell positioning and aberrant spacing of axonal fascicles in the nerve roots observed in embryos with altered ladybird function suggest that the ladybird genes might also control directed cell movements and cell-cell interactions within the developing Drosophila ventral nerve cord. PMID- 15136146 TI - A stepwise model system for limb regeneration. AB - The amphibian limb is a model that has provided numerous insights into the principles and mechanisms of tissue and organ regeneration. While later stages of limb regeneration share mechanisms of growth control and patterning with limb development, the formation of a regeneration blastema is controlled by early events that are unique to regeneration. In this study, we present a stepwise experimental system based on induction of limb regeneration from skin wounds that will allow the identification and functional analysis of the molecules controlling this early, critical stage of regeneration. If a nerve is deviated to a skin wound on the side of a limb, an ectopic blastema is induced. If a piece of skin is grafted from the contralateral side of the limb to the wound site concomitantly with nerve deviation, the ectopic blastema continues to grow and forms an ectopic limb. Our analysis of dermal cell migration, contribution, and proliferation indicates that ectopic blastemas are equivalent to blastemas that form in response to limb amputation. Signals from nerves are required to induce formation of both ectopic and normal blastemas, and the diversity of positional information provided by blastema cells derived from opposite sides of the limb induces outgrowth and pattern formation. Hence, this novel and convenient stepwise model allows for the discovery of necessary and sufficient signals and conditions that control blastema formation, growth, and pattern formation during limb regeneration. PMID- 15136147 TI - Mouse Zic5 deficiency results in neural tube defects and hypoplasia of cephalic neural crest derivatives. AB - Zic family genes encode zinc finger proteins, which are homologues of the Drosophila pair-rule gene odd-paired. In the present study, we characterized the fifth member of the mouse Zic family gene, mouse Zic5. Zic5 is located near Zic2, which is responsible for human brain malformation syndrome (holoprosencephaly, or HPE). In embryonic stages, Zic5 was expressed in dorsal part of neural tissues and limbs. Expression of Zic5 overlapped with those of other Zic genes, most closely with Zic2, but was not identical. Targeted disruption of Zic5 resulted in insufficient neural tube closure at the rostral end, similar to that seen in Zic2 mutant mice. In addition, the Zic5-deficient mice exhibited malformation of neural-crest-derived facial bones, especially the mandible, which had not been observed in other Zic family mutants. During the embryonic stages, there were delays in the development of the first branchial arch and extension of the trigeminal and facial nerves. Neural crest marker staining revealed fewer neural crest cells in the dorsal cephalic region of the mutant embryos without significant changes in their migration. When mouse Zic5 was overexpressed in Xenopus embryos, expression of a neural crest marker was enhanced. These findings suggested that Zic5 is involved in the generation of neural crest tissue in mouse development. ZIC5 is also located close to ZIC2 in humans, and deletions of 13q32, where ZIC2 is located, lead to congenital brain and digit malformations known as the "13q32 deletion syndrome". Based on both their similar expression pattern in mouse embryos and the malformations observed in Zic5-deficient mutant mice, human ZIC5 might be involved in the deletion syndrome. PMID- 15136148 TI - Pattern and morphogenesis of presumptive superficial mesoderm in two closely related species, Xenopus laevis and Xenopus tropicalis. AB - The mesoderm, comprising the tissues that come to lie entirely in the deep layer, originates in both the superficial epithelial and the deep mesenchymal layers of the early amphibian embryo. Here, we characterize the mechanisms by which the superficial component of the presumptive mesoderm ingresses into the underlying deep mesenchymal layer in Xenopus tropicalis and extend our previous findings for Xenopus laevis. Fate mapping the superficial epithelium of pregastrula stage embryos demonstrates ingression of surface cells into both paraxial and axial mesoderm (including hypochord), in similar patterns and amounts in both species. Superficial presumptive notochord lies medially, flanked by presumptive hypochord and both overlie the deep region of the presumptive notochord. These tissues are flanked laterally by superficial presumptive somitic mesoderm, the anterior tip of which also appears to overlay the presumptive deep notochord. Time-lapse recordings show that presumptive somitic and notochordal cells move out of the roof of the gastrocoel and into the deep region during neurulation, whereas hypochordal cells ingress after neurulation. Scanning electron microscopy at the stage and position where ingression occurs suggests that superficial presumptive somitic cells in X. laevis ingress into the deep region as bottle cells whereas those in X. tropicalis ingress by "relamination" (e.g., [Dev. Biol. 174 (1996) 92]). In both species, the superficially derived presumptive somitic cells come to lie in the medial region of the presumptive somites during neurulation. By the early tailbud stages, these cells lie at the horizontal myoseptum of the somites. The morphogenic pathway of these cells strongly resembles that of the primary slow muscle pioneer cells of the zebrafish. We present a revised fate map of Xenopus, and we discuss the conservation of superficial mesoderm within amphibians and across the chordates and its implications for the role of this tissue in patterning the mesoderm. PMID- 15136149 TI - Independent roles for retinoic acid in segmentation and neuronal differentiation in the zebrafish hindbrain. AB - Segmentation of the vertebrate hindbrain into rhombomeres is essential for the anterior-posterior patterning of cranial motor nuclei and their associated nerves. The vitamin A derivative, retinoic acid (RA), is an early embryonic signal that specifies rhombomeres, but its roles in neuronal differentiation within the hindbrain remain unclear. Here we have analyzed the formation of primary and secondary hindbrain neurons in the zebrafish mutant neckless (nls), which disrupts retinaldehyde dehydrogenase 2 (raldh2), and in embryos treated with retinoid receptor (RAR) antagonists. Mutation of nls disrupts secondary, branchiomotor neurons of the facial and vagal nerves, but not the segmental pattern of primary, reticulospinal neurons, suggesting that RA acts on branchiomotor neurons independent of its role in hindbrain segmentation. Very few vagal motor neurons form in nls mutants and many facial motor neurons do not migrate out of rhombomere 4 into more posterior segments. When embryos are treated with RAR antagonists during gastrulation, we observe more severe patterning defects than seen in nls. These include duplicated reticulospinal neurons and posterior expansions of rhombomere 4, as well as defects in branchiomotor neurons. However, later antagonist treatments after rhombomeres are established still disrupt branchiomotor development, suggesting that requirements for RARs in these neurons occur later and independent of segmental patterning. We also show that RA produced by the paraxial mesoderm controls branchiomotor differentiation, since we can rescue the entire motor innervation pattern by transplanting wild-type cells into the somites of nls mutants. Thus, in addition to its role in determining rhombomere identities, RA plays a more direct role in the differentiation of subsets of branchiomotor neurons within the hindbrain. PMID- 15136150 TI - Glial progenitors of the neonatal subventricular zone differentiate asynchronously, leading to spatial dispersion of glial clones and to the persistence of immature glia in the adult mammalian CNS. AB - The subventricular zone (SVZ) of the developing mammalian forebrain gives rise to astrocytes and oligodendrocytes in the neocortex and white matter, and neurons in the olfactory bulb in perinatal life. We have examined the developmental fates and spatial distributions of the descendants of single SVZ cells by infecting them in vivo at postnatal day 0-1 (P0-1) with a retroviral "library". In most cases, individual SVZ cells gave rise to either oligodendrocytes or astrocytes, but some generated both types of glia. Members of glial clones can disperse widely through the gray and white matter. Progenitors continued to divide after stopping migration, generating clusters of related cells. However, the progeny of a single SVZ cell does not differentiate synchronously: individual clones contained both mature and less mature glia after short or long intervals. For example, progenitors that settled in the white matter generated three types of clonal oligodendrocyte clusters: those composed of only myelinating oligodendrocytes, of both myelinating oligodendrocytes and non-myelinating oligodendrocytes, or of only non-myelinating cells of the oligodendrocyte lineage. Thus, some progenitors do not fully differentiate, but remain immature and may continue to cycle well into adult life. PMID- 15136151 TI - Sonic hedgehog signaling regulates Gli3 processing, mesenchymal proliferation, and differentiation during mouse lung organogenesis. AB - Lack of Sonic hedgehog (Shh) signaling, mediated by the Gli proteins, leads to severe pulmonary hypoplasia. However, the precise role of Gli genes in lung development is not well established. We show Shh signaling prevents Gli3 proteolysis to generate its repressor forms (Gli3R) in the developing murine lung. In Shh(-/-) or cyclopamine-treated wild-type (WT) lung, we found that Gli3R level is elevated, and this upregulation appears to contribute to defects in proliferation and differentiation observed in the Shh(-/-) mesenchyme, where Gli3 is normally expressed. In agreement, we found Shh(-/-);Gli3(-/-) lungs exhibit enhanced growth potential. Vasculogenesis is also enhanced; in contrast, bronchial myogenesis remains absent in Shh(-/-);Gli3(-/-) compared with Shh(-/-) lungs. Genes upregulated in Shh(-/-);Gli3(-/-) relative to Shh(-/-) lung include Wnt2 and, surprisingly, Foxf1 whose expression has been reported to be Shh dependent. Cyclins D1, D2, and D3 antibody labelings also reveal distinct expression patterns in the normal and mutant lungs. We found significant repression of Tbx2 and Tbx3, both linked to inhibition of cellular senescence, in Shh(-/-) and partial derepression in Shh(-/-); Gli3(-/-) lungs, while Tbx4 and Tbx5 expressions are less affected in the mutants. Our findings shed light on the role of Shh signaling on Gli3 processing in lung growth and differentiation by regulating several critical genes. PMID- 15136152 TI - Acetylcholinesterase function is dispensable for sensory neurite growth but is critical for neuromuscular synapse stability. AB - The enzyme acetylcholinesterase (AChE) terminates synaptic transmission at cholinergic synapses by hydrolyzing the neurotransmitter acetylcholine. In addition, AChE is thought to play several 'non-classical' roles that do not require catalytic function. Most prominent among these is facilitation of neurite growth. Here, we report that the zebrafish zieharmonika (zim) locus encodes AChE. We show that one mutant zim allele is caused by a pre-mature stop codon, resulting in a truncated protein that lacks both the catalytic site and the carboxy-terminal neuritogenic domain. To explore the 'non-classical' role of AChE, we examined embryos mutant for this allele. In contrast to previous results using a catalytic-inactive allele, our analysis demonstrates that AChE is dispensable for muscle fiber development and Rohon-Beard sensory neuron growth and survival. Moreover, we show that in the absence of AChE, acetylcholine receptor clusters at neuromuscular junctions initially assemble, but that these clusters are not maintained. Taken together, our results demonstrate that AChE is dispensable for its proposed non-classical roles in muscle fiber formation and sensory neuron development, but is crucial for regulating the stability of neuromuscular synapses. PMID- 15136153 TI - Calreticulin on the mouse egg surface mediates transmembrane signaling linked to cell cycle resumption. AB - Calreticulin, a protein best known as an endoplasmic reticulum chaperone, also is found on the extracellular plasma membrane surface of many cell types where it serves as a mediator of adhesion and as a regulator of the immune response. In this report, we demonstrate that calreticulin is present on the extracellular surface of the mouse egg plasma membrane and is increased in the perivitelline space after egg activation. The extracellular calreticulin appears to be secreted by vesicles in the egg cortex that are distinct from cortical granules. An anticalreticulin antibody binds to extracellular calreticulin on live eggs and inhibits sperm-egg binding but not fusion. In addition, engagement of cell surface calreticulin by incubation of mouse eggs in the presence of anticalreticulin antibodies results in alterations in the localization of cortical actin and the resumption of meiosis as indicated by alterations in chromatin configuration, decreases in cdc2/cyclin B1 and MAP kinase activities, and pronuclear formation. These events occur in the absence of any observable alterations in intercellular calcium. These data demonstrate that calreticulin functionally interacts with the egg cytoskeleton and can mediate transmembrane signaling linked to cell cycle resumption. These studies suggest a role for calreticulin as a lectin that may be involved in signal transduction events during or after sperm-egg interactions at fertilization. PMID- 15136154 TI - The N-terminal prodomain of sV23 is essential for the assembly of a functional vitelline membrane network in Drosophila. AB - The vitelline membrane is a specialized extracellular matrix that surrounds and protects the oocyte. Recent studies indicate that it also serves as a storage site for embryonic pattern determinants. sV23, a major vitelline membrane protein, is essential for the morphogenesis of the vitelline membrane as sV23 protein null mutants lay flaccid, infertile eggs. By analyzing a series of sV23 mutant transgenes in the sV23 protein null genetic background, we have shown that sV23 is secreted as a proprotein in functional excess and that C- and N-terminal prodomains are removed successively, following its deposition in the extracellular space. Although a target site for subtilisin-like convertases is essential for N-terminal processing, N-terminal processing is not necessary for the assembly of a functional vitelline membrane layer. While C-terminal truncations were tolerated, the removal of N-terminal sequences lead to the production of flaccid, infertile eggs with a soluble, rather than insoluble, vitelline membrane network. We propose that the hydrophobic N-terminal prodomain plays an early and essential role in aligning molecules within the vitelline membrane network, much like hydrophobic domains within elastin drive the assembly and alignment of molecules within elastin-based extracellular matrices. PMID- 15136155 TI - Microsecond freeze-hyperquenching: development of a new ultrafast micro-mixing and sampling technology and application to enzyme catalysis. AB - A novel freeze-quench instrument with a characteristic <> of 137 +/- 18 micros is reported. The prototype has several key features that distinguish it from conventional freeze-quench devices and provide a significant improvement in time resolution: (a) high operating pressures (up to 400 bar) result in a sample flow with high linear rates (up to 200 m s(-1)); (b) tangential micro-mixer with an operating volume of approximately 1 nl yields short mixing times (up to 20 micros); (c) fast transport between the mixer and the cryomedium results in short reaction times: the ageing solution exits the mixer as a free-flowing jet, and the chemical reaction occurs "in-flight" on the way to the cryomedium; (d) a small jet diameter (approximately 20 microm) and a high jet velocity (approximately 200 m s(-1)) provide high sample-cooling rates, resulting in a short cryofixation time (up to 30 micros). The dynamic range of the freeze-quench device is between 130 micros and 15 ms. The novel tangential micro-mixer efficiently mixes viscous aqueous solutions, showing more than 95% mixing at eta < or = 4 (equivalent to protein concentrations up to 250 mg ml(-1)), which makes it an excellent tool for the preparation of pre-steady state samples of concentrated protein solutions for spectroscopic structure analysis. The novel freeze-quench device is characterized using the reaction of binding of azide to metmyoglobin from horse heart. Reaction samples are analyzed using 77 K optical absorbance spectroscopy, and X-band EPR spectroscopy. A simple procedure of spectral analysis is reported that allows (a) to perform a quantitative analysis of the reaction kinetics and (b) to identify and characterize novel reaction intermediates. The reduction of dioxygen by the bo3-type quinol oxidase from Escherichia coli is assayed using the MHQ technique. In these pilot experiments, low-temperature optical absorbance measurements show the rapid oxidation of heme o3 in the first 137 micros of the reaction, accompanied by the formation of an oxo-ferryl species. X-band EPR spectroscopy shows that a short-living radical intermediate is formed during the oxidation of heme o3. The radical decays within approximately 1 ms concomitant with the oxidation of heme b, and can be attributed to the PM reaction intermediate converting to the oxoferryl intermediate F. The general field of application of the freeze-quench methodology is discussed. PMID- 15136156 TI - Forced rotation of Na+-driven flagellar motor in a coupling ion-free environment. AB - Rotational characteristics of Na+-driven flagellar motor in the presence and absence of coupling ion were analyzed by electrorotation method. The motor rotated spontaneously in the presence of Na+, and the rotation accelerated or decelerated following the direction of the applied external torque. The spontaneous motor rotation was inhibited by removal of external Na+, however, the motor could be forcibly rotated by relatively small external torque applied by the electrorotation apparatus. The observed characteristic of the motor was completely different from that of ATP-driven motor systems, which form rigor bond when their energy source, ATP, is absent. The internal resistance of the flagellar motor increased significantly when the coupling ion could not access the inside of the motor, suggesting that the interaction between the rotor and the stator is changed by the binding of the coupling ion to the internal sites of the motor. PMID- 15136157 TI - Structural characterization of a binuclear center of a Cu-containing NO reductase homologue from Roseobacter denitrificans: EPR and resonance Raman studies. AB - Aerobic phototrophic bacterium Roseobacter denitrificans has a nitric oxide reductase (NOR) homologue with cytochrome c oxidase (CcO) activity. It is composed of two subunits that are homologous with NorC and NorB, and contains heme c, heme b, and copper in a 1:2:1 stoichiometry. This enzyme has virtually no NOR activity. Electron paramagnetic resonance (EPR) spectra of the air-oxidized enzyme showed signals of two low-spin hemes at 15 K. The high-spin heme species having relatively low signal intensity indicated that major part of heme b3 is EPR-silent due to an antiferromagnetic coupling to an adjacent CuB forming a Fe Cu binuclear center. Resonance Raman (RR) spectrum of the oxidized enzyme suggested that heme b3 is six-coordinate high-spin species and the other hemes are six-coordinate low-spin species. The RR spectrum of the reduced enzyme showed that all the ferrous hemes are six-coordinate low-spin species. Nu(Fe-CO) and nu(C-O) stretching modes were observed at 523 and 1969 cm(-1), respectively, for CO-bound enzyme. In spite of the similarity to NOR in the primary structure, the frequency of nu(Fe-CO) mode is close to those of aa3- and bo3-type oxidases rather than that of NOR. PMID- 15136158 TI - The human mitochondrial KATP channel is modulated by calcium and nitric oxide: a patch-clamp approach. AB - ATP-sensitive potassium channels of the inner mitochondrial membrane (mtKATP) are blocked by ATP. They are suggested to be involved in protective mechanisms such as ischemic preconditioning (IPC). Here we identify this channel type for the first time in a human cell line (Jurkat cells). Vesicles of the inner mitochondrial membrane (mitoplasts) were prepared by hypoosmotic shock. Single channel currents were measured by means of the patch-clamp technique. We identified an outward-rectifying channel with a slope conductance of 15 and 82 pS at negative and positive potentials, respectively. The block by 5-hydroxydecanoic acid and inhibition by ATP characterize this channel as the mtKATP channel. ATP also increased the frequency of events within the burst. This effect was modulated by the Ca2+-bath concentration. We also show that the human mtKATP channel is a direct target for nitric oxide that blocked the channel activity. Although the molecular structure of this channel type is still unknown, its characterization as an outward-rectifying channel and modulation by calcium ions and nitric oxide may help to elucidate its functional significance, which possibly implicates a role in cell survival after IPC. PMID- 15136159 TI - Functional reconstitution of a maltose ATP-binding cassette transporter from the thermoacidophilic gram-positive bacterium Alicyclobacillus acidocaldarius. AB - The thermoacidophilic gram-positive bacterium Alicyclobacillus acidocaldarius grows at 60 degrees C and pH 2-3. The organism can utilize maltose and maltodextrins as energy source that are taken up by an ATP-binding cassette (ABC) import system. Genes encoding a maltose binding protein, MalE, and two membrane integral subunits, MalF and MalG, are clustered on the chromosome but a malK gene translating into a cognate ATPase subunit is lacking. Here we report the cloning of malK from genomic DNA by using the msiK gene of Streptomyces lividans as a probe. Purified MalK exhibited a spontaneous ATPase activity with a Vmax of 0.13 micromol Pi/min/mg and a Km of 330 microM that was optimal at the growth temperature of the organism. Coexpression of malK, malF and malG in Escherichia coli resulted in the formation of a complex that could be coeluted from an affinity matrix after solubilization of membranes with dodecylmaltoside. Proteoliposomes prepared from the MalFGK complex and preformed phospholipid vesicles of A. acidocaldarius displayed a low intrinsic ATPase activity that was stimulated sevenfold by maltose-loaded MalE, thereby indicating coupling of ATP hydrolysis to substrate translocation. These results provide evidence for MalK being the physiological ATPase subunit of the A. acidocaldarius maltose transporter. Moreover, to our knowledge, this is the first report on the functional reconstitution of an ABC transport system from a thermophilic microorganism. PMID- 15136160 TI - The two forms of the S(2) state multiline signal in Photosystem II: effect of methanol and ethanol. AB - The characteristic Mn hyperfine 'multiline' signal exhibited in the S2 state of the oxygen-evolving complex (OEC) complex of Photosystem II (PSII) has been shown to be heterogeneous in character. In this study, we have explored the effects that influence the proportions of the two forms of the S2 state multiline signal present in any sample. The narrow form of the signal is lost upon storage (weeks) at 77 K, whereas the broad form remains. In particular, we explore the roles of ethanol and methanol as well as effects of the second turnover of the enzyme on storage of the sample at 77 K. We find that in samples containing methanol, the narrow form may predominate upon the first flash, but the broad form predominates on the fifth flash and also in samples containing ethanol. PMID- 15136161 TI - Methyl-CpG binding domain column chromatography as a tool for the analysis of genomic DNA methylation. PMID- 15136162 TI - Toward pyrosequencing on surface-attached genetic material by use of DNA-binding luciferase fusion proteins. AB - Mutation detection and single-nucleotide polymorphism genotyping require screening of large samples of materials and therefore the importance of high throughput DNA analysis techniques is significant. Pyrosequencing is a four enzyme bioluminometric DNA sequencing technology based on the sequencing-by synthesis principle. Currently, the technique is limited to simultaneous analysis of 96 or 384 samples. Earlier, attempts to increase the sample capacity were made using micromachined filter chamber arrays where parallel analyses of nanoliter samples could be monitored in real time. We have developed a strategy for specific immobilization of the light-producing enzyme luciferase to the DNA template within a reaction chamber. By this approach, luciferase is genetically fused to a DNA-binding protein (Klenow polymerase or Escherichia coli single stranded DNA-binding (SSB) protein) and to a purification handle (Z(basic)). The proteins are produced in E. coli and purified using cation and anion exchange chromatography with removal of Z(basic). The produced proteins have been analyzed using an assay for complete primer extension of DNA templates immobilized on magnetic beads detected by pyrosequencing chemistry. Results from these experiments show that the proteins bind selectively to the immobilized DNA and that their enzymatic domains were active. Z(basic)-SSB-luciferase produced the highest signal in this assay and was further exploited as enzymatic reagent for DNA sequencing. PMID- 15136163 TI - Polarographic behaviors of diclofenac sodium in the presence of dissolved oxygen and its analytical application. AB - The polarographic response characteristics of diclofenac sodium were investigated in 0.25 M HAc-NaAc (pH 5.0) supporting electrolyte in the absence and the presence of dissolved oxygen. The results demonstrate that the reduction peak at ca. -1.10 V is a catalytic hydrogen wave after deaeration, and the reduction peak in the presence of dissolved oxygen is a so-called parallel catalytic hydrogen wave. Based on the parallel catalytic hydrogen wave, a novel method has been proposed for the determination of diclofenac sodium by single-sweep polarography. The calibration curve is linear in the range 1.2 x 10(-7)-2.6 x 10(-6)M and the detection limit is 6.0 x 10(-8)M. The proposed method is applied to the direct determination of diclofenac sodium in tablet forms and biological samples. PMID- 15136164 TI - Cell-based screen of HMG-CoA reductase inhibitors and expression regulators using LC-MS. AB - We present an integrated consolidation of previously reported methods for screening hydroxymethylglutaryl-coenzyme A reductase (HMGR) inhibitors in 96-well microtiter plates with rapid workup using established mammalian cell lines and liquid chromatography-mass spectrometry analysis. Inhibitors as well as expression regulators of HMGR (inducers or repressors) can be screened. To validate the method, three competitive inhibitors of HMGR (lovastatin, simvastatin, and atorvastatin), as well as a potent sterol repressor of HMGR synthesis (25-hydroxycholesterol), were assayed on two cell lines: HepG2, a human hepatic derived cell line, and L cells, a subline of NCTC clone 929 mouse fibroblasts. The direct inhibition of HMGR by statins, induction of HMGR synthesis by the same statins following incubation with the cells, and repression of HMGR synthesis by 25-hydroxycholesterol were confirmed. PMID- 15136165 TI - Use of cellular glucose-6-phosphate dehydrogenase for cell quantitation: applications in cytotoxicity and apoptosis assays. AB - A fluorescence-based microplate assay was developed to quantify cell death based upon the measurement of glucose-6-phosphate dehydrogenase (G6PD) activity. G6PD is a cytosolic enzyme and leaks from cells when plasma membrane integrity is compromised. In this assay, cell death is measured by correlating the activity of extracellular G6PD to the reduction of resazurin to the fluorescent product, resorufin, via a coupled-enzyme reaction. The coupled-enzyme reaction permits rapid signal amplification from small amounts of G6PD, an advantage over assays based on resazurin alone. This assay is rapid, nontoxic, and amenable to high throughput screening. The assay has a Z' factor of 0.78. PMID- 15136166 TI - Resolution of Cys and Lys labeling of alpha-crystallin with site-sensitive fluorescent 3-hydroxyflavone dye. AB - Ratiometric fluorescent probes based on 3-hydroxyflavone (3HF) are highly sensitive tools for studying polarity, hydration, electronic polarizability, and electrostatics in different microheterogeneous systems, including protein molecules. In the present work, a reactive derivative of 3HF, 6-bromomethyl-4' diethylamino-3-hydroxyflavone, recently synthesized in our group, was applied to label covalently bovine lens alpha-crystallin. The labeling of SH and NH(2) groups are clearly distinguished by spectroscopic criteria. We observe that the NH(2) labeling creates the positive charge in the proximity to fluorophore, which results in strong internal Stark effect producing the shift in excitation spectrum by ca. 15 nm. Analysis of excitation-dependent fluorescence spectra allows separation of the emission profiles of these SH- and NH(2)-labeled species. Applying recently developed multiparametric analysis of the obtained emission spectra, we described the physicochemical properties of the sites of SH and NH(2) labeling in alpha-crystallin. The site of SH labeling has medium-low polarity (dielectric constant, epsilon = 4.9 +/- 0.9) is protic, and does not contain proximal aromatic residues (according to the obtained refractive index, n = 1.41 +/- 0.14). The site of NH(2) labeling is also of medium-low polarity. The novel label due to its two-wavelength ratiometric response and high sensitivity to the type of labeling may offer new possibilities in the studies of structure, dynamics, and interactions of proteins by probing their SH- and NH(2)-labeling sites. PMID- 15136167 TI - Increased sample capacity for genotyping and expression profiling by kinetic polymerase chain reaction. AB - We fabricated and evaluated high-throughput kinetic thermal cyclers with 768 reaction capacity for kinetic polymerase chain reaction (kPCR)-based genotyping and kinetic reverse transcription (kRT)-PCR-based transcript quantitation. The system uses dye-based detection with ethidium bromide and a single DNA polymerase based PCR or RT-PCR assay. Allele-specific detection of the two most common hereditary hemochromotosis mutant alleles, C282Y and H63D, was reliably measured by kPCR using human DNA templates as low as 10 genome equivalents per assay. Transcript profiling was performed for 16 yeast transcripts ranging in intracellular abundance over four orders of magnitude. Standard deviations of the PCR cycle threshold values determined from multiple kRT-PCR assays in three different instruments ranged from 0.11 to 0.97 PCR cycles and were reproducible, transcript specific, and instrument independent. The effects of the sin3, gal11, and snf2 knockout mutations on expression of 385 yeast genes were evaluated by kRT-PCR and compared to published values determined by high-density oligonucleotide array and/or microarray analysis for snf2 and sin3. The 768 reaction kinetic thermalcyclers, each with a capacity for more than a half million assays per year, are well suited to genomics applications such as single nucleotide polymorphism/disease association studies and genomewide transcription profiling where high sensitivity and accuracy are required. PMID- 15136168 TI - Development of high-throughput screens for discovery of kinesin adenosine triphosphatase modulators. AB - Kinesins are a group of related molecular motor proteins that have great potential as targets for antimitotic drug development. We have developed two novel assays, one end-point and one kinetic, that are useful for the discovery and optimization of kinesin modulators. Both assays measure inorganic phosphate (Pi) generated by microtubule-activated kinesin adenosine triphosphatase activity. The assays were validated using the mitotic Eg5 kinesin-specific inhibitor, monastrol. A panel of nine kinesin motor domain proteins, representing 8 of the 14 classes of kinesins, was screened. The coefficient of variation for both assays was determined to be 4-14% depending on the panel member. Using the Eg5 kinetic assay with monastrol the IC50 value was 12 microM, which agrees well with previously published results. Two other closely related mitotic kinesins (AnBimC and MKLP1) were found to have IC50 values in the millimolar range. The other panel members (kinesin heavy chain, chromokinesin KIF4A, KIF3C, CENP-E, MCAK, and KIFC3) were not significantly inhibited by millimolar levels of monastrol. It is anticipated that screening of the nine-member panel of kinesins in these assays will serve as a platform for the discovery and development of specific kinesin modulators. PMID- 15136169 TI - An Alizarin red-based assay of mineralization by adherent cells in culture: comparison with cetylpyridinium chloride extraction. AB - Alizarin red S (ARS) staining has been used for decades to evaluate calcium-rich deposits by cells in culture. It is particularly versatile in that the dye can be extracted from the stained monolayer and assayed. This study describes a sensitive method for the recovery and semiquantification of ARS in a stained monolayer by acetic acid extraction and neutralization with ammonium hydroxide followed by colorimetric detection at 405 nm. This method was three times more sensitive than an older method involving cetylpyridinium chloride (CPC) extraction and resulted in a better signal to noise ratio, especially for weakly stained monolayers. The assay facilitates detailed inspection of mineralization by phase microscopy and semiquantification of the entire monolayer by extraction and quantification. The sensitivity of the assay is improved by the extraction of the calcified mineral at low pH and, since the mineral is already stained in a quantitative manner, there is no requirement for an additional colorimetric quantification step. Furthermore, the linear range is much wider than those of conventional assays for calcium, making dilutions of mineral extracts prior to measurement unnecessary. It has a wide range of potential uses including tumor characterization, mesenchymal stem cell evaluation, and osteogenic compound screening. Although more labor intensive than CPC extraction, the protocol is more sensitive and yields more reliable results for weakly mineralizing samples. PMID- 15136170 TI - Tuning the redox and enzymatic activity of glucose oxidase in layered organic films and its application in glucose biosensors. AB - Glucose oxidase was embedded in organic films through a layer-by-layer approach, where the enzyme demonstrated significantly enhanced electron-transfer reactivity and finely tuned enzymatic activity. An unmediated, reagentless glucose biosensor was accordingly prepared with two polyethylenimine/glucose oxidase bilayers modified pyrolytic graphite electrode. A calibration linear range of glucose was 0.5-8.9 mM with a detection limit of 50 microM and sensitivity of 0.76 microA mM( 1). PMID- 15136171 TI - Algorithm-assisted elucidation of disulfide structure: application of the negative signature mass algorithm to mass-mapping the disulfide structure of the 12-cysteine transforming growth factor beta type II receptor extracellular domain. AB - The power of an algorithm-driven method for interpreting disulfide mass-mapping data is demonstrated in the context of determining the disulfide structure of the extracellular domain of the transforming growth factor beta type II receptor, a 14-kDa cystinyl protein containing 12 cysteines in the form of six disulfide bonds. The disulfide mass-mapping methodology is based on partial reduction and cyanylation-induced cleavage of the cystinyl protein. Because the multiplicity of possible disulfide structures that must be considered grows rapidly with the number of cysteines, as does the difficulty in physically isolating each of the partially reduced and cyanylated isoforms of the analyte, manual data interpretation for disulfide mapping a cystinyl protein containing more than eight cysteines becomes unmanageable. Recently, we introduced the concept of a "negative signature mass algorithm" (NSMA) to determine the disulfide structure of a cystinyl protein by processing an input of its amino acid sequence and mass spectral data from analysis of its associated cyanylation-induced cleavage products. Here, we present experimental results to validate the NSMA concept. A key advantage of the NSMA, in addition to convenience and automation, is its capacity to interpret mass spectra from mixtures of cyanylation-induced cleavage fragments without separating the partially reduced isoforms of the cystinyl protein and without knowledge of the extent of partial reduction. PMID- 15136172 TI - Probing structural features of water-insoluble proteins by front-face fluorescence. AB - Front-face fluorescence was used to assess some structural features of proteins in wheat flours, taking advantage of the fact that this technique allowed a direct approach to a number of systems in which most proteins cannot be solubilized without using conditions that strongly modify the structure of proteins in the original material. In this study, we addressed structural changes ensuing from solvation and from mechanical deformation of proteins in wheat flour. Solvation of proteins in the systems under investigation was monitored by taking fluorescence emission spectra of the protein tryptophans (or of protein bound 1,8-anilino-naphthalene-sulfonate added in appropriate amounts) in mixtures of increasing water content. In separate experiments, changes in overall protein surface hydrophobicity were estimated by titrating flours-mixed with appropriate amounts of water to a dough-like consistency-with increasing concentrations of the fluorescent hydrophobic probe, 1,8-anilino-naphthalene-sulfonate. This approach allowed concomitant determination of the overall binding capacity and of the apparent affinity for the probe of proteins. To test the usefulness of the structural information obtained from these approaches, they were also applied to characterize dough at different levels of mechanical stress, prepared from either wheat flour or semolina. PMID- 15136173 TI - Measurement of the affinity and cooperativity of annexin V-membrane binding under conditions of low membrane occupancy. AB - We developed a method for measuring the binding affinity of annexin V for phospholipid vesicles and cells at very low levels of membrane occupancy. The annexin V-117 mutant was labeled with fluorescein iodoacetamide on its single N terminal cysteine residue; binding to phospholipid vesicles containing phosphatidylserine (PS) and 2% rhodamine-phosphatidylethanolamine was measured by fluorescence quenching due to resonance energy transfer; binding to cells with exposed PS was measured by fluorometry after elution of bound protein. The equilibrium constant was calculated as a function of the midpoint of the calcium titration curve, the Hill coefficient, and the concentration of membrane binding sites. Calcium titrations at very low ratios of protein to membrane revealed Hill coefficients of approximately 8 for both vesicles and cells, far higher than previously measured, but as the protein-membrane ratio was increased above 3% of maximum membrane occupancy, the value of the Hill coefficient progressively decreased to a limiting value of about 2. High Hill coefficients were also observed for measurements performed at different ionic strengths and with membrane PS content varied over the range from 20 to 50%. This method allows the accurate determination of the affinity and cooperativity of annexin V-membrane binding and will be useful for the evaluation of modified annexin V derivatives intended for diagnostic and therapeutic applications. PMID- 15136174 TI - Comparative proteomic analysis of planktonic and immobilized Pseudomonas aeruginosa cells: a multivariate statistical approach. AB - The protein maps of Pseudomonas aeruginosa cells from two natural (attached) and one artificial (gel-entrapped) immobilized-cell (IC) systems, together with their free (suspended) counterparts, were compared after incubation for 18 or 48 h in a minimal salt medium. Principal component analysis (PCA) was used to interpret the variations in protein spot densities that were observed on electropherogram obtained by two-dimensional electrophoresis (2-DE). PCA of the 2-DE data, a matrix of 933 rows (observations, i.e., spot density values) and 12 columns (variables, i.e., incubation conditions), in which observations were standardized horizontally, extracted four principal components (PCs) accounting for 78.75% of the variability in the protein expression profiles. PC1 opposed the two modes of growth (planktonic and immobilized) while PC2 discriminated between the incubation times of free cell cultures. The incubation conditions of ICs, including the immobilization procedure (entrapment vs attachment) and the nature of the biofilm substratum, were fairly separated in PC3xPC4. The dependence of the protein patterns on the cell immobilization process was further illustrated by the identification of a number of peptides whose amount remained unchanged or was altered in ICs compared to free bacteria. These results reinforce the topical assertion that bacteria in the immobilized state display a specific physiological behavior but also question the existence of a unique IC phenotype. PMID- 15136175 TI - Screening assays for cholinesterases resistant to inhibition by organophosphorus toxicants. AB - Methods to measure resistance to inhibition by organophosphorus toxicants (OP) for mutants of butyrylcholinesterase (EC 3.1.1.8; BChE) and acetylcholinesterase (EC 3.1.1.7; AChE) enzymes were devised. Wild-type cholinesterases were completely inhibited by 0.1 mM echothiophate or 0.001 mM diisopropylfluorophosphate, but human BChE mutants G117H, G117D, L286H, and W231H and snake AChE mutant HFQT retained activity. Tissues containing a mixture of cholinesterases could be assayed for amount of G117H BChE. For example, the serum of transgenic mice expressing human G117H BChE contained 0.5 microg/ml human G117H BChE, 2 microg/ml wild-type mouse BChE, and 0.06 microg/ml wild-type mouse AChE. The oligomeric structure of G117H BChE in the serum of transgenic mice was determined by nondenaturing gel electrophoresis followed by staining for butyrylthiocholine hydrolysis activity in the presence of 0.1 mM echothiophate. Greater than 95% of the human G117H BChE in transgenic mouse serum was a tetramer. To visualize the distribution of G117H BChE in tissues of transgenic mice, sections of small intestine were treated with echothiophate and then stained for BChE activity. Both wild-type and G117H BChE were in the epithelial cells of the villi. These assays can be used to identify OP-resistant cholinesterases in culture medium and in animal tissues. PMID- 15136176 TI - Removal of lipid contaminants by organic solvents from oilseed protein extract prior to electrophoresis. PMID- 15136177 TI - Time-resolved luminescence anisotropy-based detection of immunoglobulin G using long-lifetime Ru(II) complex-labeled protein A. PMID- 15136178 TI - Anchor primer associated problems in differential display reverse transcription polymerase chain reaction. PMID- 15136179 TI - Casein provides protection against proteolytic artifacts. PMID- 15136180 TI - DNA mutagenesis using T4 DNA polymerase and DpnI restriction endonuclease. PMID- 15136181 TI - Removal of sputtered gold from specimens after scanning electron microscope examination. PMID- 15136182 TI - Detection of phospholipase D on solid materials. PMID- 15136183 TI - An unmediated H2O2 biosensor based on the enzyme-like activity of myoglobin on multi-walled carbon nanotubes. PMID- 15136184 TI - An introduction to assessing anxiety in child and adolescent multiethnic populations: challenges and opportunities for enhancing knowledge and practice. AB - Recognizing that researchers and clinicians are increasingly faced with assessing, treating, and studying ethnically, racially, and culturally diverse populations, one question is whether the appropriate tools exist. This special section aspires to advance the available assessment strategies that are appropriate for the treatment, prevention, and research of diverse children and adolescents with anxiety symptoms and disorders. This introduction presents an overview of anxiety and its disorders among multiethnic youth and identifies emerging challenges and opportunities in the assessment and treatment of anxiety symptoms and disorders among Asian American and Pacific Islander, Hispanic/Latino, African American, and European American children and adolescents within a sociocultural context. PMID- 15136185 TI - Temperament, anxiety, and depression: comparisons across five ethnic groups of children. AB - Extending analyses from a large school-based sample of children and adolescents (N = 1,126; Chorpita, 2002), this study examined anxiety and depression assessment with 5 ethnic groups in Hawaii. Ethnic differences in anxiety and depression symptoms, along with 2 temperamental characteristics-negative affectivity (NA) and positive affectivity (PA)-were examined within the context of the tripartite model of anxiety and depression (Clark & Watson, 1991). Minimal between-group differences in temperamental characteristics were found. Good fit was found for a multisample model relating NA and PA to anxiety and depressive dimensions consistently across all groups. However, significant mean level differences among ethnic groups were found for several specific anxiety dimensions, suggesting that factors outside of the tripartite model explain observed ethnic differences. Further support for the generalizability of the tripartite model was also found. PMID- 15136186 TI - Clinical phenomenology, somatic symptoms, and distress in Hispanic/Latino and European American youths with anxiety disorders. AB - This study compared clinic-anxious Hispanic/Latino and European American youths (ages 6 to 17 years old) along sociodemographic and clinical variables. Groups were relatively similar, although significant differences emerged as a function of ethnocultural and language choice (English, Spanish) used during the assessment. Within the English language choice group, Cuban American (CA) youths reported somatic symptoms as less distressing than non-Cuban American Hispanic/Latino (non-CA/HL) youths. Conversely, within the Spanish language choice group, CA youths reported somatic symptoms as more distressing than non CA/HL youths. Also, parents in the European American and CA groups reported their youths as having less somatic symptoms than parents in the non-CA/HL group. Implications of findings are discussed, particularly regarding possible cultural significance of somatic symptoms. PMID- 15136187 TI - Anxiety reporting and culturally associated interpretation biases and cognitive schemas: a comparison of Mexican, Mexican American, and European American families. AB - This study examined whether Mexican (n = 53), Mexican American (n = 50), and European American (n = 51) children differed in their reporting of anxiety symptoms and whether parental influence and specific cognitive schemas associated with Mexican culture were related to differences in anxiety reporting. As expected, Mexican and Mexican American children reported significantly more physiological and worry symptoms than the European American children. Mexican and Mexican American children endorsed collectivism as a cultural value more strongly than European American children, and the Mexican children evidenced greatest use of social strategies reflecting simpatia. In family discussions of ambiguous, potentially anxiety-arousing situations, Mexican and Mexican American parents verbalized a greater percentage of somatic interpretations than the European American parents. Results indicate potential linkages between cultural values, socialization practices, and anxiety reporting. PMID- 15136188 TI - Assessing anxiety sensitivity in inner-city African American children: psychometric properties of the childhood anxiety sensitivity index. AB - This study examined the psychometric properties of the Childhood Anxiety Sensitivity Index (CASI; Silverman et al., 1991) in a sample of urban African American elementary school children. One hundred forty-four 4th- and 5th-grade children completed the CASI as part of a larger project. In contrast to prior research with community samples, CASI total scores were higher and means did not differ by sex. Internal consistency (alpha =.82), 2-week test-retest reliability (r =.80), and convergent and divergent validity were good and consistent with prior research. The factor structure of the CASI differed from the structure found with White children. Results of exploratory factor analyses suggest two factors: physical concerns and mental incapacitation concerns. Implications for assessing anxiety and anxiety sensitivity among African American children are discussed. PMID- 15136189 TI - Assessment and treatment of socially phobic children: a cross cultural comparison. AB - In this study we examined the psychopathology and behavioral treatment of White and African American preadolescent children with social phobia. The comprehensive assessment strategy, including semistructured diagnostic interviews, clinician ratings of impairment, behavioral observations, parental ratings, and self-report inventories, did not reveal differences in symptomatic presentation between African American and White children. Whereas all children improved from pre- to posttreatment, there were no significant differences based on race. The results are discussed in terms of the applicability of models 0of social phobia and treatment outcome across these 2 ethnic groups. PMID- 15136190 TI - Assessment of anxiety and depression in Asian American youth. AB - This article reviews the current state of research on the assessment of anxiety and depression in Asian American children and adolescents. Contrary to lay perceptions of Asian Americans as a "model minority," research indicates that rates of depression and anxiety among Asian American adults are comparable to those found among European American populations. Given that Asian American populations also underutilize mental health services, the need to assess and treat anxiety and depression in Asian American youth is critical. This commentary includes directions for future research as well as clinical intervention. PMID- 15136191 TI - Are somatic symptoms and related distress more prevalent in Hispanic/Latino youth? Some methodological considerations. AB - This article comments on the current status of the anxiety literature involving Latino children and adolescents. As the 2 articles that focus on Hispanic/Latino youth in this special section independently found somatic symptoms to be more prevalent in Latino youth than other racial/ethnic groups (Pina & Silverman, this issue; Varela et al., this issue), this commentary discusses methodological considerations for guiding future anxiety research and makes suggestions for clinicians who treat Latino children and adolescents. PMID- 15136192 TI - Orphans no more: a commentary on anxiety and African American youth. AB - Conducting research on anxiety disorders among African American children and adolescents, prior to the early 1990s was considered an "orphan area" of psychology. The articles in this section, first authored by a new generation of researchers, appear to set a direction for future research. The findings underscore the importance of including culturally relevant variables in the assessment of African American children's anxiety and its disorders. PMID- 15136193 TI - Halting the development of conduct problems in head start children: the effects of parent training. AB - We examined parent and child moderators of outcome, program engagement effects, and predictors of engagement in the Incredible Years Parent Training Program. Head Start classrooms (N = 882 children) were randomly assigned to an intervention condition (that received the Incredible Years program) or to a control condition (that received usual Head Start services). Structural equation modeling (SEM) was used to model the effects of the training program on child outcomes. The analyses showed differential program effects depending on children's initial levels of conduct problems and mothers' initial levels of critical parenting. Children with high baseline levels of conduct problems and children of mothers with high initial levels of critical parenting benefited most from the program. Changes in children's conduct problems were also related to maternal engagement in the program and to intervention mothers' success at reducing their critical parenting. PMID- 15136194 TI - Development and validation of a gender-balanced measure of aggression-relevant social cognition. AB - This study examined the psychometric properties of the Social-Cognitive Assessment Profile (SCAP), a gender-balanced measure of social information processing (SIP) in a sample of 371 (139 girls, 232 boys) 2nd- to 4th-grade children. The SCAP assesses 4 dimensions of SIP (Inferring Hostile Intent, Constructing Hostile Goals, Generating Aggressive Solutions, and Anticipating Positive Outcomes for Aggression) in the context of peer conflict involving relational and overt provocation. Confirmatory factor analyses indicated that the 4 latent factors provided a good fit to the data for girls and boys and for African American and non-African American children. Regression analyses in which teacher and peer evaluations of aggression and peer evaluations of social competencies were regressed on each of the 4 SCAP scales supported the test's convergent and discriminant validity. These results suggest that the SCAP is an easily administered and brief measure of SIP that is appropriate for racially diverse populations of elementary boys and girls. PMID- 15136195 TI - Parent and teacher agreement on child behavior checklist items in a sample of preschoolers from low-income and predominantly African American families. AB - In this study we examined parent and teacher agreement at the item level of the newly revised Child Behavior Checklist/1 1/2-5 (CBCL/1 1/2-5) and Caregiver Teacher Report Form/1 1/2-5 (CTRF/1 1/2-5) in 505 preschool children from low income and predominantly African American families. Parents generally rated more children as having problem behaviors than did teachers. Lack of agreement between parents and teachers at the item level was indicated by low correlation coefficients, kappa values, and co-identification of children with specific behavior problems. High levels of parent-teacher agreement were obtained only when rank orders of problem behaviors were compared. No significant sex effect was found on parent and teacher agreement at the item level. Problem behaviors most often indicated by parents and teachers reflected the roles and responsibilities of parents and teachers in the home and school settings. Our findings suggest informant and setting specificity of parent- and teacher identified problem behaviors in young children. PMID- 15136196 TI - The structure and correlates of perfectionism in African American children. AB - This study examined the structure and symptom correlates of perfectionism in a sample of 6th-grade, urban, African American children using the Child and Adolescent Perfectionism Scale (CAPS; Flett, Hewitt, Boucher, Davidson, & Munro, 2000). Confirmatory factor analysis showed inadequate fit of the original subscales. Exploratory factor analysis suggested 3 dimensions of perfectionism: A Socially Prescribed and a Self-Oriented-Critical factor were described as maladaptive, whereas a Self-Oriented-Striving factor was proposed as adaptive. Cross-sectional correlations among the perfectionism dimensions and symptoms of psychopathology fit well with the adaptive versus maladaptive model. In addition, Socially Prescribed Perfectionism showed a robust predictive relation to 7th grade internalizing symptoms, especially depression, in boys. Results are discussed in the context of the cultural and socioeconomic characteristics of this African American sample and suggestions for future research are provided. PMID- 15136197 TI - Applying depression-distortion hypotheses to the assessment of peer victimization in adolescents. AB - This study examined whether adolescents' depressive symptoms and aggressive behavior were associated with discrepancies between self- and peer-reports of peer victimization experiences. A sample of 203 10th-grade adolescents completed self-report measures of victimization and depressive symptoms as well as peer nominations of victimization and aggression. Residual scores were computed as a measure of discordance between peer- and self-reported peer victimization. Adolescents' aggressive behavior was associated with underestimations of peer victimization on self-reported measures, as compared to peer-reports, whereas depressive symptoms were associated with overestimations of peer victimization on self-report, as compared to peer-reports. Different patterns of findings were revealed for different forms of victimization (overt, relational, reputational) and by gender. Findings have implications for studies of adolescent peer victimization using multiple reporters and suggest that adolescents with high levels of depressive symptoms may be vulnerable to misperceptions of their social experiences among peers. PMID- 15136198 TI - Antisocial behavior and affiliation with deviant peers. AB - We examined the associations among gender, antisocial behavior, and peer-group affiliation in a high-risk sample of 401 homeless and matched housed adolescents (139 boys and 262 girls). The Diagnostic Interview Schedule for Children (Version 2.3, 1991; Costello, Edelbrock, Kalas, Kessler, & Klaric, 1982) yielded 2 measures of adolescent antisocial behavior: symptoms of conduct disorder and substance abuse or dependence. Various deviant behaviors of friends were assessed based on adolescent self-report. Results indicated that, for both boys and girls, having many deviant peers was associated with more antisocial behavior, regardless of the number of boys in the peer network. Furthermore, findings suggest that the relation between number of deviant peers and antisocial behavior may be stronger for boys and homeless adolescents than for girls and housed adolescents, respectively. The results of prior studies indicating that antisocial behavior is a function of affiliation with male peers may be due to the higher frequencies of maladaptive behaviors evidenced in boys in normative samples. PMID- 15136199 TI - Longitudinal relations between parenting and child adjustment in young children. AB - We studied the predictive relations between reports of parenting behavior on the one hand and academic achievement and reported behavior problems of young children on the other hand. Data were gathered for 352 children and their parents from kindergarten to 2nd grade. The results indicated that in the academic domain, low supportive and high controlling parenting practices were modestly related to poor subsequent math achievement. Children's externalizing and attention problem behavior was clearly predictive of high levels of control in mothers and low levels of support in fathers. The combination of high parental support and control was especially associated with high levels of problem behavior. However, when previous parenting and child adjustment were taken into account, the magnitude of the predictive power of parenting for child adjustment, and of child adjustment for parenting, remained limited. PMID- 15136200 TI - Innovative treatment for children with serious emotional disturbance: preliminary outcomes for a school-based intensive mental health program. AB - This article describes the development, implementation, and preliminary evaluation of a school-based Intensive Mental Health Program (IMHP) for 50 children (42 boys, 8 girls) with severe, early-onset, serious emotional disturbances (SED). Eighty-four percent of the children showed clinically significant improvement in overall functioning as measured by the Child and Adolescent Functional Assessment Scale (CAFAS). Child functioning at home and school, behavior toward others, regulation of moods and emotions, self-harm, and problems in thinking improved significantly. Results provide initial support for the IMHP as a promising approach to serving the needs of children with SED. PMID- 15136201 TI - Knowledge of social anxiety disorder relative to attention deficit hyperactivity disorder among educational professionals. AB - Social anxiety disorder (SAD), the 3rd most common psychiatric disorder in the United States, follows a chronic and unremitting course, often resulting in severe impairments in multiple areas of functioning. Despite a typical age of onset in early adolescence, the disorder is rarely recognized and treated in adolescent populations. Given its early age of onset, school professionals are arguably in the best position to detect symptoms of SAD and to provide appropriate referrals for assessment and intervention. This study examined the knowledge that teachers, school counselors, and school psychologists have of SAD in relation to their knowledge of a prototypical externalizing disorder, attention deficit hyperactivity disorder (ADHD). The results suggest that all 3 groups of educational professionals are surprisingly unfamiliar with adolescent SAD. PMID- 15136202 TI - Bias in self-perceptions and internalizing and externalizing problems in adjustment during early adolescence: a prospective investigation. AB - We investigated bias in self-perceptions of competence (relative to parent ratings) for family, school, and peer domains as predictors of adjustment problems among 139 young adolescents over a 1-year period using a prospective design. Regressions examined measures of bias at Time 1 (T1) as predictors of ratings of internalizing and externalizing problems at Time 2 (T2), controlling for T1 adjustment ratings. For the family domain, curvilinear trends were found. Follow-up analyses revealed that for this domain both negative bias (self perceptions less favorable than parent ratings) and positive bias (self perceptions more favorable than parent ratings) predicted greater internalizing and externalizing problems as rated by youth, parents, and teachers. For the peer domain, higher scores on the measure of bias predicted greater internalizing and externalizing problems as rated by teachers. These findings are consistent with the view that accuracy in self-perceptions of competence can have important implications across multiple domains of development. PMID- 15136203 TI - Do deviant peer associations mediate the contributions of self-esteem to problem behavior during early adolescence? A 2-year longitudinal study. AB - We investigated deviant peer associations as a mediator of the influences of general and peer-oriented self-esteem on problem behavior using data from a 2 year longitudinal study of 350 young adolescents. Measures of problem behavior included substance use (alcohol use, smoking) and antisocial behavior (fighting, stealing). Using latent growth curve modeling and covariance structure analysis, an extension of a model proposed by DuBois et al. (2002) was evaluated for each type of problem behavior. Findings revealed that lower general self-esteem and greater peer orientation in self-esteem each predicted deviant associations with peers and that deviant peer associations, in turn, were associated with higher levels and rates of change in problem behavior. Deviant peer associations mediated the associations of general and peer-oriented self-esteem with levels and rates of change in problem behavior such that direct paths from self-esteem to problem behavior generally were nonsignificant. PMID- 15136204 TI - Social construction: vistas in clinical child and adolescent psychology. AB - We explore here the potentials of a social constructionist orientation to knowledge for research and clinical practice. Dialogues on social construction emphasize the communal origins of knowledge. They stress the cultural basis of knowledge claims, the significance of language, the value saturation of all knowledge, and the significance of relationships as opposed to individuals. An initial illustration of constructionism in action centers on adolescent risk behavior. Such behavior is often constructed negatively within popular writings and the social science and thus ignores the meaning of such actions to the adolescents themselves. Discourse analysis indicates that for adolescents risky behavior serves important functions of enhancing group solidarity and establishing positive identity. A second illustration, exploring the implications of constructionism for therapy, places a strong emphasis on the therapist as a collaborator in the building of meaning. Traditional investments in diagnosis and treatment are replaced with the collaborative creation of new possibilities for action. PMID- 15136205 TI - Parental mental disorder and children's functioning: silence and communication, stigma and resilience. AB - In this article I argue that narrative accounts of the effects of parental mental disorder (particularly mood disturbance) on children's functioning may provide unprecedented windows on crucial issues of silence, stigmatization, diagnosis, etiology, and treatment, which can mutually inform empirical research efforts. In publishing a book-length account of the life of my father, The Years of Silence are Past (Hinshaw, 2002b), I departed from my usual academic writings and gave voice to a complex and troubling life history, marked by misdiagnosed bipolar disorder and misguided treatment that lasted 40 years. Herein, I focus on the ramifications of this narrative for scientific and clinical efforts in the field, including the still-prevalent stigmatization and silence surrounding mental disorder, the great need for accurate diagnosis and responsive treatment, the complex and transactional web of biological and environmental risk factors that exist for mental disturbance, the blending of inner experience with social realities regarding symptom content, the resilience and strength that can accompany mental disorder, and the ways in which parents with mental disturbance can promote active communication with their offspring. Narrative accounts may be particularly important in reducing silence and in framing the next generation of empirical research questions; blending qualitative and quantitative methods may help address crucial goals related to child functioning in the presence of parental mental disorder. PMID- 15136206 TI - Stressors and child and adolescent psychopathology: measurement issues and prospective effects. AB - This article reviews existing research on the association between stressors and symptoms of psychopathology in children and adolescents with a focus on measurement issues and prospective effects. The first half of the article focuses on the measurement of stressors, emphasizing checklists and interviews. Available measures of stressful experiences are reviewed and critiqued. Results of this review reveal both substantial progress (i.e., development of valid stressor assessment tools) and remaining problems (i.e., inconsistent measurement across studies). The second half of this article reviews studies that have tested for prospective associations between stressors and symptoms of psychopathology in children and adolescents. Studies that have examined the prospective effects of recent or prior stressors on current psychological symptoms, while controlling for prior psychological symptoms, are reviewed. Results overall suggest that stressors predict changes in rates of symptoms of psychopathology in children and adolescents over time. Results also suggest that symptoms of psychopathology predict changes in rates of stressors over time. Implications of these findings are that conclusive evidence now exists for the importance of stressors in the development of child and adolescent psychopathology. PMID- 15136207 TI - Parents' beliefs about condoms and oral contraceptives: are they medically accurate? AB - CONTEXT: Parents are encouraged to be the primary sex educators for their children; however, little is known about the accuracy of parents' views about condoms and oral contraceptives. METHODS: Telephone surveys using validated measures provided data on beliefs about the effectiveness, safety and usability of condoms and the pill among 1,069 parents of 13-17-year-olds in Minnesota and Wisconsin in 2002. Pearson chi-square tests and multivariate logistic regression models were used to compare beliefs according to sex, age, race, religion, education, income and political orientation. RESULTS: Substantial proportions of parents underestimated the effectiveness of condoms for preventing pregnancy and sexually transmitted diseases (STDs). Only 47% believed that condoms are very effective for STD prevention, and 40% for pregnancy prevention. Fifty-two percent thought that pill use prevents pregnancy almost all the time; 39% thought that the pill is very safe. Approximately one-quarter of parents thought that most teenagers are capable of using condoms correctly; almost four in 10 thought that most teenagers can use the pill correctly. Fathers tended to have more accurate views about condoms than mothers did; mothers' views of the pill were generally more accurate than fathers'. Whites were more likely than nonwhites to hold accurate beliefs about the pill's safety and effectiveness; conservatives were less likely than liberals to hold accurate views about the effectiveness of condoms. CONCLUSION: Campaigns encouraging parents to talk with their teenagers about sexuality should provide parents with medically accurate information on the effectiveness, safety and usability of condoms and the pill. PMID- 15136208 TI - Attitudes and intentions of future health care providers toward abortion provision. AB - CONTEXT: Induced abortion is one of the most common procedures performed among women in the United States. However, 87% of all counties had no abortion provider in 2000, and little is known about the attitudes and intentions of future health care providers, including advanced clinical practitioners, regarding abortion provision. METHODS: During March 2002, first- and second-year students in health sciences programs (i.e., medicine, physician assistant and nursing) at the University of Washington were anonymously surveyed. Univariate, bivariate and multivariate analyses were used to determine students' attitudes and intentions regarding provision of abortion services. RESULTS: Of the 312 students who completed the survey, 70% supported the availability of legal abortion under any circumstances. Thirty-one percent intended to provide medical abortion in their practice, and 18% planned to offer surgical abortion. Fifty-two percent of all respondents agreed that advanced clinical practitioners should be able to provide medical abortion, and 37% agreed that they should be able to provide surgical abortion services; however, greater proportions of advanced clinical practitioners (45-83%) than of medical students (21-43%) expressed such support. Sixty-four percent of all respondents were willing to attend a program whose curriculum requires abortion training. CONCLUSIONS: Although it may not be possible to require abortion training for every future health care provider, making abortion a standard part of clinical training would provide opportunities for future physicians and advanced clinical practitioners, and would likely ameliorate the abortion provider shortage. PMID- 15136209 TI - Acceptability of the vaginal diaphragm among current users. AB - CONTEXT: Interest in the diaphragm has been growing, in part because it is a female-controlled method that might protect against HIV and other sexually transmitted diseases (STDs). A better understanding of diaphragm acceptability is needed. METHODS: In 2001-2002, female members of a managed care organization were interviewed by telephone. The 215 participants, aged 19-49, who reported diaphragm use during the past three months were asked about their experience with the method and background characteristics. Characteristics associated with women's satisfaction with and consistent use of the diaphragm were identified through multiple logistic regression analysis. RESULTS: Most participants had a low risk for HIV and other STDs. The mean duration of diaphragm use was 8.5 years. Although only 42% of participants reported consistent use in the past three months, most were satisfied with the method (79%) and planned to use it at next vaginal intercourse (85%). Satisfied users had significantly higher diaphragm use self-efficacy and more positive perceptions of the method than those not satisfied. Consistent use was significantly associated with older age and having had some college education rather than none. More than half of women cited dissatisfaction with previous methods (72%) and provider recommendation (61%) as moderately to extremely important in their decision to begin diaphragm use. When asked what they would change about the diaphragm, 32% mentioned concerns related to inserting or removing it. CONCLUSIONS: From an acceptability point of view, the diaphragm appears to be a viable candidate for a female controlled method for prevention of HIV and other STDs. Our findings have important implications for the reintroduction of the traditional diaphragm and development of new diaphragm-like products. PMID- 15136210 TI - U.S. insurance coverage of contraceptives and the impact of contraceptive coverage mandates, 2002. AB - CONTEXT: In 1993, coverage of reversible contraception by U.S. health insurance plans was extremely low. Little is known about how coverage has changed since then, particularly in response to state mandates. METHODS: In 2001-2002, a nationally representative sample of 205 health care insurers responded to a survey about coverage of reproductive health care services in "typical" employment-based managed care plans (excluding self-insured plans). Data were analyzed to compare coverage in states with and without contraceptive coverage mandates, and to show trends in coverage between 1993 and 2002. RESULTS: In 2002, almost every reversible contraceptive service and supply studied was covered by at least 89% of typical plans; 86% of plans covered the five leading prescription methods (the diaphragm, one- and three-month injectables, the IUD and oral contraceptives). Coverage of each contraceptive service and supply studied was higher in 2002 than in 1993 (78-97% vs. 32-59%). Plans in states with mandates were significantly more likely to cover the five leading prescription methods (87 92%, depending on type of plan) than were those designed locally in states without mandates (47-61%). Between 1993 and 2002, state mandates were estimated to account for 30% and 40% of the increase in coverage of oral contraceptives and the three-month injectable, respectively. CONCLUSIONS: Coverage of reversible contraception--and by extension, choice within a range of covered methods--has increased substantially since 1993, in part because of state mandates. This state by-state approach, however, has inherent limitations that can best be dealt with at the federal level. PMID- 15136211 TI - The HIPAA privacy rule and adolescents: legal questions and clinical challenges. PMID- 15136212 TI - Moving forward in myeloma research. PMID- 15136214 TI - Adverse effects and benefits of two years of anagrelide treatment for thrombocythemia in chronic myeloproliferative disorders. AB - BACKGROUND AND OBJECTIVES: Although anagrelide is widely used in the treatment of thrombocythemia in myeloproliferative diseases, there is currently limited information on the efficacy and toxicity of its long-term use. This prospective study investigated clinical toxicity and efficacy of anagrelide during two years of treatment. DESIGN AND METHODS: A multicenter, open, phase II study of anagrelide treatment was performed by the Swedish Myeloproliferative Disorder Study Group. The study included 60 patients with thrombocythemia due to myeloproliferative disease, 42 with essential thrombocythemia (ET), 17 with polycythemia vera (PV) and one with myelofibrosis (MF). RESULTS: Complete response (CR), defined as a platelet count <400x10(9)/L in symptomatic patients and < 600x10(9)/L in asymptomatic patients was achieved in 67% of the patients and partial response (PR) in 6%. The response rate was higher in patients with ET than in those with PV (p = 0.05). Primary treatment failure occurred in 27% due to lack of efficacy at a tolerable dose (n=13) or insufficient platelet response without side effects (n=3). In addition, another 14 patients withdrew from treatment before the end of the two-year period due to side effects. Side effects included palpitations (70%), headache (52%), nausea (35%), diarrhea or flatulence (33%), edema (22%) and fatigue (23%). Patients and doctors rated their satisfaction with the anagrelide treatment on a 10-grade scale from 7.6 at 3 months to >9 at 24 months. After two years, 50% (n=30) of the patients continued anagrelide treatment. INTERPRETATION AND CONCLUSIONS: Side effects and toxic discontinuation rates were higher than in previous studies, probably because this is the first long-term prospective study of the feasibility and toxicity of anagrelide treatment. Nevertheless, anagrelide is a valuable alternative for treatment of thrombocythemia in myeloproliferative disorders for patients who tolerate the drug well. PMID- 15136215 TI - Prognostic impact of early response to induction therapy as assessed by multiparameter flow cytometry in acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: Early response after induction therapy is an independent prognostic factor in acute myeloid leukemia (AML). We improved the identification of this parameter by implementing multiparameter flow cytometry to quantify bone marrow cells carrying leukemia-associated immunophenotypes (LAIP). DESIGN AND METHODS: In 106 uniformly treated patients flow cytometric analyses were performed at diagnosis and one week after induction therapy (day 16). The log-difference between LAIP-positive cells on day 1and day 16 (LD16) was determined for each patient. RESULTS: The LD16 (median, 2.11; range, -0.37 to 4.20) was significantly correlated to CR rate, event-free survival (EFS), overall survival (OS), and relapse-free survival (RFS). Separation of patients by the median LD16 resulted in significant differences in CR rate (81% vs. 51%, p=0.002), EFS (53% at 2 years vs. median 2.8 months, p<0.0001), 2-year OS (58% vs. 43%, p=0.0133), and 2-year RFS (65% vs. 30%, p=0.0037). Multivariate analysis revealed that LD16 was an independent prognostic parameter for CR rate, EFS, and RFS. INTERPRETATION AND CONCLUSIONS: Flow cytometric evaluation of early response may serve as a new response parameter in AML. It may be used for development of risk-adapted therapies. High-risk patients can be identified early after the first induction therapy and assigned alternative and salvage treatment strategies. PMID- 15136216 TI - Acquired potential N-glycosylation sites within the tumor-specific immunoglobulin heavy chains of B-cell malignancies. AB - BACKGROUND AND OBJECTIVES: Among B-cell malignancies, follicular lymphomas (FL) more frequently show acquired, potential N-glycosylation sites (AGS) within tumor specific immunoglobulin. The aim of this study was to extend this observation and to evaluate the pattern of presentation of AGS within five different forms of B cell lymphoma. DESIGN AND METHODS: We sequenced the tumor-specific immunoglobulin heavy chain variable region fragment, including complementarity-determining regions 2 and 3, of forty-seven consecutive patients with a B-cell malignancy enrolled in idiotype vaccine clinical trials. This sequencing approach is known to allow the identification of most AGS. We then statistically analyzed differences in presentation pattern, in terms of tumor histology, immunoglobulin isotype, AGS location and amino acid composition. RESULTS: All twenty-four FL cases presented with at least one AGS, whereas the vast majority of four B-cell lymphoma types other than FL did not. The non- FL group of tumors included four cases of Burkitt's lymphoma, six of diffuse large cell lymphoma, seven mantle cell lymphomas and six small lymphocytic lymphomas. Most IgM-bearing follicular lymphoma cases featured their AGS within complementarity-determining region 2, as opposed to those bearing an IgG, which mostly displayed the AGS within complementarity-determining region 3. The vast majority of AGS located within either complementarity-determining region ended with a serine residue, whereas those located within framework regions mostly featured threonine as the last amino acid residue. INTERPRETATION AND CONCLUSIONS: In our series, all cases of FL had AGS within their tumor-specific immunoglobulin heavy chain variable regions. In contrast, most B-cell malignancies other than FL did not. Further studies are warranted in order to establish the possible meaning of these findings in terms of disease pathogenesis, their diagnostic value in doubtful cases and their potential implications for immunotherapy. PMID- 15136217 TI - Patients with CD45 negative multiple myeloma receiving high-dose therapy have a shorter survival than those with CD45 positive multiple myeloma. AB - BACKGROUND AND OBJECTIVES: CD45 is a critical regulator of signaling threshold in immune cells. There are clinical and animal studies suggesting that the CD45 negative phenotype is the phenotype of progressive multiple myeloma (MM). The aims of this study were to confirm this hypothesis and to test the prognostic value of CD45 expression in newly diagnosed MM patients. DESIGN AND METHODS: In a retrospective study of 95 newly diagnosed MM patients treated with high dose therapy we used 4-color flow cytometry to determine CD45 expression and correlated the immunophenotipic data with clinical data. RESULTS: Thirty of 95 patients (31.5%) lacked CD45 expression at diagnosis. The CD45 phenotype significantly affected the overall survival (OS) of the patients, like the most common presenting prognostic parameters analyzed including b-2-microglobulin, age and 14q32 translocations. CD45 negative MM patients had a significantly worse OS than did CD45 positive cases of MM: 28.7% cumulative survival at 4 years, median 42 months vs not reached; p = 0.004. Furthermore, CD45 remained the only parameter adversely affecting OS in multivariate analysis. INTERPRETATION AND CONCLUSIONS: The CD45 negative phenotype could reflect the phenotype of progressive disease in relation to the intrinsic malignancy of the MM clone. Indeed, CD45 negative myeloma cells appear to have a greater capacity to circulate, disseminate and clone as well as being less sensitive to apoptosis. PMID- 15136218 TI - A low serum level of soluble tumor necrosis factor receptor p55 predicts response to thalidomide in advanced multiple myeloma. AB - BACKGROUND AND OBJECTIVES: Thalidomide modulates the production of tumor necrosis factor (TNF-alpha). Soluble TNF receptors, TNFR p55 and TNFR p75, modify TNF alpha activity. In this study, we explored the relation between soluble TNF receptors and outcome in patients with advanced multiple myeloma treated with thalidomide. DESIGN AND METHODS: The levels of soluble TNF receptor p55 and p75 were assessed in serum from 34 myeloma patients with relapsed or refractory disease before starting thalidomide treatment. Serial measurements were performed for 16 patients in serum collected during treatment. RESULTS: The pre-treatment serum level of soluble TNFR p55 in thalidomide responders was significantly lower than that in non-responders (median 1.75 ng/mL (range 1.19-2.84) vs. 2.79 ng/mL (1.36-5.51), p=0.004). The levels of p55 declined significantly during treatment. The levels of p75 showed the same pattern as p55, but the differences were not significant. The median survival of myeloma patients with pre-treatment levels of p55 < 2.79 ng/mL was 404 days; the median survival of patients with pre-treatment levels >or= 2.79 ng/mL was shorter (65 days, log-rank test p=0.02). INTERPRETATION AND CONCLUSIONS: We conclude that soluble TNFR p55 is an adverse prognostic factor in myeloma patients with relapsed or refractory disease treated with thalidomide. Patients with a low pre-treatment level of this receptor have a better response rate and a longer overall survival. PMID- 15136219 TI - Levels of minimal residual disease detected by quantitative molecular monitoring herald relapse in patients with multiple myeloma. AB - BACKGROUND AND OBJECTIVES: Detection of minimal residual disease (MRD) has helped to improve the treatment of patients with leukemia. At present MRD testing in patients with multiple myeloma (MM) is not applied as a standard diagnostic or prognostic method. DESIGN AND METHODS: Immunoglobulin heavy chain (IgH) polymerase chain reaction (PCR) using patient-specific TaqMan probes together with LightCycler technology was performed to quantify minimal residual disease in MM. Relative levels of clonotypic cells were assessed as IgH/2beta-actin ratios with a sensitivity of 10(-4) to 10(-5). RESULTS: Following stem cell transplantation, a significant reduction of clonotypic cells was observed in bone marrow (BM) and peripheral blood (PB) samples of 11 patients, comparing pre treatment values with those of best response (median: 13% to 0.09% and 0.03% to 0%, respectively). In 5 patients with ongoing clinical remission IgH/2beta-actin ratios remained stable at a low level, while in 6 patients an increase to 2% in BM and 0.4% in PB was associated with progression of the disease. In 4 of these 6 patients the increase of clonotypic cells in PB was detectable a median of 3 months (range: 0.5-6) before relapse. Furthermore, time-to-progression of patients with pre-transplantation IgH/2b-actin ratios > 0.03% in BM was significantly shorter than that of patients with lower MRD levels. INTERPRETATION AND CONCLUSIONS: MRD in patients with MM can be quantified reliably using TaqMan chemistry adapted to the LightCycler system. Residual tumor cell levels before transplantation as well as results of sequential molecular monitoring are predictive of relapse. PMID- 15136220 TI - Sequential analysis of biochemical markers of bone resorption and bone densitometry in multiple myeloma. AB - BACKGROUND AND OBJECTIVES: Bone lesions often occur in multiple myeloma (MM), but no tests have proven useful in identifying patients with increased risk. Bone marker assays and bone densitometry are non-invasive methods that can be used repeatedly at low cost. This study was performed to evaluate these methods in predicting bone events in MM patients. DESIGN AND METHODS: Thirty newly diagnosed MM patients were enrolled. Serum C-terminal telopeptide (ICTP) and urinary N terminal telopeptide (NTx) of collagen I were measured for assessment of bone resorption, and serum C-terminal (PICP) and N-terminal (PINP) propeptides of procollagen I, bone-specific alkaline phosphatase, and osteocalcin were measured to estimate bone formation. Dual energy X-ray absorptiometry (DEXA) was used to assess bone mineral density (BMD) of the lumbar spine, hip, and whole body. Serum and urine samples were collected every 6 weeks, DEXA-scans performed every 3 months, and skeletal radiographs were done every 6 months as well as when indicated. RESULTS: Serum ICTP and urinary NTx were predictive of progressive bone events. Markers of bone formation, bone mineral density assessments, and M component measurements were less informative. In Cox analysis, ICTP showed the highest predictive value, but should be replaced with NTx in patients with nephropathy. Pretreatment low lumbar BMD was predictive of early vertebral fractures. INTERPRETATION AND CONCLUSIONS: Sequential DEXA-scans showed heterogeneous local BMD changes, and our data do not support routine use of sequential DEXA-scans. However, lumbar DEXA-scans at diagnosis can identify patients with increased risk of early vertebral collapses. Sequential analyses of serum ICTP and urinary NTx are useful for monitoring bone damage. PMID- 15136221 TI - Granulocyte colony-stimulating factor increases expression of adhesion receptors on endothelial cells through activation of p38 MAPK. AB - BACKGROUND AND OBJECTIVES: Granulocyte colony-stimulating factor (G-CSF) is specific for the granulocytic cell line, although receptors for this cytokine have been found in other cell types including endothelial cells. These observations prompted us to investigate the potential effect of G-CSF on the endothelium. DESIGN AN METHODS: Endothelial cell monolayers were exposed to G-CSF to evaluate: i) signal transduction mechanisms, ii) expression of adhesion receptors at the cell surface, and iii) leukocyte adhesion on EC monolayers. RESULTS: Exposure of human umbilical vein endothelial cells (EC) in culture to G CSF resulted in the activation of the signal transduction pathways JAK/STAT (JAK 1, STAT-1 and STAT-3) and RAS/MAPK (MAPK p42/44 and p38 MAPK). We also observed significantly increased expression of the adhesion receptors, E-selectin (ELAM 1), vascular endothelial cell adhesion molecule-1 and intracelleular adhesion molecule-1 at the cell surface in response to G-CSF, increases that were followed by an augmented adhesion of leukocytes on the previously exposed EC monolayers. These effects were blocked by the presence of SB203580, a p38 MAPK inhibitor, by U0126, a MAPK p42/44 inhibitor, and by inhibiting the G-CSF receptor with a specific antibody. INTERPRETATION AND CONCLUSIONS: Our results demonstrate that G CSF increases the expression of adhesion receptors on EC, promoting leukocyte adhesion. This effect seems to be triggered by the signaling events that follow receptor binding. Results from experiments using specific inhibitors suggest that activation of p38 MAPK is required to promote expression of adhesion receptors in endothelial cells and the recruitment of leukocytes in response to G-CSF. PMID- 15136222 TI - Thromboprophylaxis practice patterns in two Western Australian teaching hospitals. AB - BACKGROUND AND OBJECTIVES: Evidence-based international guidelines recommend that all patients undergoing elective hip or knee arthroplasty receive thromboprophylaxis with low-molecular-weight heparin or adjusted-dose warfarin. Our objective was to determine what proportion of patients undergoing elective hip or knee arthroplasty actually receive recommended thromboprophylaxis according to international guidelines. DESIGN AND METHODS: We performed a prospective cohort study of 396 consecutive patients undergoing elective hip or knee arthroplasty between 1 May and 30 October, 2002. We collected baseline data, surgical and anesthetic details and recorded use of thromboprophylaxis and episodes of venous thromboembolism that occurred within 3 months of surgery. RESULTS: The mean age of the patients was 69.4 years (SD 11.5 years), and 62.2% (95% CI: 57.3 to 66.9%) were female. Hip arthroplasty was performed in 39.4% (34.6 to 44.2%) and knee arthroplasty in 57.1% (52.2 to 61.9%). Recommended thromboprophylaxis with low-molecular-weight heparin or warfarin was administered to 51.5% (46.6 to 56.4%). Objectively diagnosed venous thromboembolism occurred in 5.3% (3.3 to 8.0%) of patients; 3.5% (1.9 to 5.9%) of events occurred during hospitalization and 1.8% (0.7 to 3.6%) occurred following discharge from hospital. There was no significant reduction in the incidence of venous thromboembolism among patients treated with recommended thromboprophylaxis compared with those who did not but this is not a randomized comparison and is potentially confounded by the indication for treatment. INTERPRETATION AND CONCLUSIONS: Current thromboprophylaxis practice at our institutions falls substantially short of national and international guidelines. The reasons for low thromboprophylaxis use should be further explored and strategies for change implemented in order to optimize clinical practice. PMID- 15136223 TI - Recommended reporting format for flow cytometry diagnosis of acute leukemia. AB - Although flow cytometry is increasingly used as a tool to diagnose hematologic malignancies, the reporting format of acute leukemia immunodiagnosis is still imprecise and sometimes vague, often reflecting old guidelines. Thus, the purpose of the present work was to make the reporting format for the immunological diagnosis of acute leukaemia easy and clear to understand. This work represents part of a more articulated series of technical guidelines that the Italian Society for Cytometry (GIC) is currently processing. Thirteen separate recommendations, covering all aspects of an acute leukemia cytometry report, are listed. According to our suggestions, the report must contain clear statements about: 1. demographic identification of patient; 2. identification of the hospital or division sending the sample; 3. type of specimen (bone marrow aspirate, peripheral blood, other biological fluids); 4. timing of observation (first diagnosis or follow-up); 5. diagnostic hypothesis made by the sender; 6. list of antigens and type of immunofluorescence analysis carried out; 7. absolute number of cells in the sample; 8. quality of the sample, in terms of viability; 9. general description of the gating procedure; 10. immunophenotype of blast cells; 11. description of cells surrounding blasts; 12. diagnostic conclusions; 13. definition of an antigen panel (when applicable) for the detection of minimal residual disease. As an example of a final report we present a case of acute myeloid leukaemia with t(8;21) translocation; in filling this report, we followed all the 13 points of the checklist described in the paper. PMID- 15136224 TI - Allogeneic hematopoietic stem cell transplantation for metastatic breast cancer. AB - The prognosis is poor and the options are limited for patients with metastatic breast cancer (MBC), especially for those patients who have previously received taxanes and anthracyclines; treatment strategies are primarily palliative. Murine models have demonstrated that allogeneic T cells are capable of eliciting graft versus-tumor (GVT) effects against breast cancer, inhibiting growth of breast cancer cell lines in vivo, providing the rationale to pursue allogeneic adoptive cellular therapy as a strategy to treat MBC. However, the clinical application of allogeneic hematopoietic stem cell transplantation (alloHSCT) was limited by concerns over toxicity and unproven efficacy. The development of non myeloablative (a.k.a. reduced-intensity) conditioning regimens, which have less treatment-related mortality but preserve the T-cell mediated GVT effects, led to increased investigation of alloHSCT in MBC. Early reports of non-myeloablative alloHSCT indicate that a clinical GVT effect against breast cancer does exist. The responses, observed in 20-40% of patients, appear to be associated with the development of complete donor lymphoid chimerism and may be delayed. In its current form, alloHSCT by itself is unlikely to result in complete eradication of MBC; however, it may serve as a therapeutic platform to complement and enhance the effects of existing cytotoxic therapies and immunotherapies (e.g. trastuzumab), as well as therapies under development (e.g. vaccines). Current data on alloHSCT for MBC should be interpreted cautiously and carefully used for the design of future studies to fully determine the clinical efficacy of this form of adoptive cellular therapy in MBC. PMID- 15136225 TI - Comparison of efficiency of ex vivo expansion of whole blood, mononuclear cells and purified CD34+ cells from human umbilical cord blood. AB - We used a dual-chamber culture system separated by a dialysis membrane to test the efficiency of expansion of whole cord blood and cell fractions. We found that expansion of progenitor cells was more efficient from whole blood than from purified CD34+ or partially purified mononuclear cell fractions. PMID- 15136226 TI - Mass spectrometry and DNA sequencing are complementary techniques for characterizing hemoglobin variants: the example of hemoglobin J-Oxford. AB - Liquid chromatography-electrospray ionization-mass spectrometry (MS) allows the characterization of most hemoglobin variants and can sometimes be a useful tool to narrow down DNA sequencing analysis. As an example, we report a case of hemoglobin variant J-Oxford, characterized by MS and DNA sequencing analysis. PMID- 15136227 TI - A new cross-over region for hemogloboin-Lepore-Hollandia. AB - Hb Lepore-Hollandia, identified in a Thai patient, was found to be due to a new cross-over between IVS1-nt 42 and nt 56 of the delta- and beta-globin genes. This differs from a previous case which crossed-over between codon 22 and IVS1-nt 16. Two independent homologous unequal cross-over events account for these two Lepore Hollandia genes. PMID- 15136228 TI - e6a2 BCR-ABL transcript in chronic myeloid leukemia: is it associated with aggressive disease? AB - We describe the fourth case of e6a2 BCR-ABL transcript in a patient with chronic myeloid leukemia (CML), using reverse transcriptase polymerase chain reaction (RT PCR) and sequencing analysis. The clinical and hematologic features and the aggressive course of disease in our patient and in the others reported in literature lead us to hypothesize that this atypical rearrangement may be associated with a worse prognosis. PMID- 15136229 TI - No mutations in the GATA-1 gene detected in patients with acquired essential thrombocythemia. AB - Mutations in the GATA-1 gene have been identified in patients with familial macrothrombocytopenia and Down's syndrome patients with a transient myeloproliferative disorder and/or acute megakaryoblastic leukemia. We screened this gene in 46 patients with essential thrombocythemia and identified only a common single nucleotide polymorphism that is unlikely to be of pathological significance. PMID- 15136230 TI - Arsenic trioxide in the treatment of advanced acute promyelocytic leukemia. AB - Eleven patients with advanced APL were treated with ATO (0.15 mg/Kg daily). Eight (73%) achieved molecular CR, but 5 relapsed, 1 died in molecular CR, 1 was lost to follow-up and 1 is still alive in CR after allogeneic transplantation. We suggest that ATO may be effective also in advanced APL, but given the short CR, it seems indicated only in patients eligible for transplant procedures. PMID- 15136231 TI - 5'-(3')-nucleotidase mRNA levels in blast cells are a prognostic factor in acute myeloid leukemia patients treated with cytarabine. AB - We analyzed cytosolic 5'-(3')-nucleotidase (dNT-1) mRNA expression by quantitative polymerase chain reaction at diagnosis in leukemic blasts from 114 patients with acute myeloid leukemia (AML) treated with ara-C. Our results show that low dNT-1 mRNA expression in leukemic blasts at diagnosis is correlated with a worse clinical outcome and suggest that this enzyme may have a role in sensitivity to ara-C in AML patients. PMID- 15136232 TI - Differentiating agents + low-dose chemotherapy in the management of old/poor prognosis patients with acute myeloid leukemia or myelodysplastic syndrome. AB - 13-cis retinoic acid + (OH)2 vitamin D3 + low-dose 6-thioguanine and cytarabine were tested in 26 patients with acute myeloid leukemia (AML) and in 4 patients with myelodysplastic syndrome (MDS) (median age 72.5), ineligible for standard chemotherapy. The response rate was 50%, with 27% complete remission. The median survival of the whole group and responders was 7.5 (1-47+) and 16.5 months (3.5 47+), respectively. PMID- 15136233 TI - Prolonged molecular remission after autologous stem cell transplantation in relapsed acute promyelocytic leukemia. AB - Six patients with relapsed acute promyelocytic leukemia (APL) received autologous stem cell transplantation (ASCT) in second (n=5) or fourth (n=1) molecular remission with a molecularly negative graft. After a median follow-up of 33 months from ASCT, 5 patients are alive in molecular remission and one died 27 months after autograft from refractory relapse. PMID- 15136234 TI - Analysis of fatal intracranial hemorrhage in 792 acute leukemia patients. AB - Forty-one of 792 acute leukemia patients suffered fatal intracranial hemorrhage (FICH). Acute promyelocytic leukemia was the most common subtype. Achievement of complete remission in AML was significantly influenced by FICH. FICH accounts for about half of deaths from hemorrhage and this proportion has not changed despite improvements in leukemia management. PMID- 15136235 TI - Bcl-6 gene mutations in primary cutaneous B-cell lymphomas. AB - We analyzed mutations in the 5' non-coding region of the BCL-6 gene in 46 cases of primary cutaneous B-cell lymphomas (PCBCL), using a polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) method. The results indicate that PCBCL display a low frequency of mutations and support a marginal zone B cell origin for most of these neoplasms. PMID- 15136236 TI - Effect of statins, smoking and obesity on progression of monoclonal gammopathy of undetermined significance: a case-control study. AB - Interleukin-6 (IL-6) and C-reactive protein (CRP, a surrogate marker for IL-6) are important in monoclonal gammopathy of undetermined significance (MGUS) and myeloma. Smoking and obesity may elevate CRP levels, while statins decrease CRP levels. A case-control study in 200 MGUS patients found that statin use, smoking history and obesity do not affect MGUS progression. PMID- 15136237 TI - Xenobiotic gene polymorphisms and susceptibility to multiple myeloma. AB - Genetic variations in the activity of xenobiotic enzymes may predict susceptibility to multiple myeloma (MM). In a case-control study, 90 Australian Caucasians with MM had significantly higher incidences of GST T1 null, PON1 BB and NAT2 slow acetylation genotypes, but no difference in polymorphism frequencies for GST M1, NAT1, and CYP1A1 when compared to 205 controls. PMID- 15136238 TI - Additional transplantation of ex vivo generated megakaryocytic cells after high dose chemotherapy. AB - The additional transplantation of ex vivo generated hematopoietic (post) progenitor cells represents a possible approach to ameliorate high-dose chemotherapy induced cytopenia. We investigated the feasibility of the large scale expansion and transplantation of autologous megakaryocytic cells in four patients with advanced solid tumors. Up to 1,460x10(6) ex vivo generated cells were administered without adverse effects but no clear cut effect on platelet recovery was observed. PMID- 15136239 TI - Clinical efficacy and prediction of response to granulocyte transfusion therapy for patients with neutropenia-related infections. AB - We investigated the efficacy of transfusing granulocytes into 32 patients with severe neutropenia-related infections and the factors that predict response to this therapy. Our findings suggest that granulocyte transfusion therapy is useful for treating neutropenic patients with fungal infections and that 99mTc-HMPAO granulocyte scintigraphy can be used to predict response to granulocyte transfusion therapy. PMID- 15136240 TI - In vivo priming with granulocyte colony-stimulating factor possibly enhances the effect of gemtuzumab-ozogamicin in acute myeloid leukemia: results of a pilot study. AB - Eight elderly patients with relapsed or refractory acute myeloid leukemia were treated sequentially with recombinant human granulocyte colony-stimulating factor with rhG-CSF and Mylotarg. Priming with rhG-CSF in vivo induced an increase in the proportion of CD33+ cycling blasts. Four patients (50% ) achieved a complete remission, 2 patients had a partial remission and the other 2 were resistant. PMID- 15136241 TI - Fatal immune-mediated pancytopenia and a TRALI-like syndrome associated with high titers of recipient-type antibodies against donor-derived peripheral blood cells after allogeneic bone marrow transplantation following dose reduced conditioning. AB - Pancytopenia occurring after bone marrow transplantation is a rare complication. A 47 year old patient with progression of multiple myeloma after standard therapy received an allogeneic marrow graft from a matched unrelated donor. The non myeloablative conditioning regimen consisted of fludarabine, cyclophosphamide, rabbit anti-thymocyte globulin and total body irradiation. GVHD prophylaxis consisted of cyclosporine. Neutrophil engraftment was as expected and the patient was discharged without signs of acute GvHD. On day +34 the patient presented with clinical and laboratory findings consistent with severe pancytopenia. Antibodies against red cells, platelets, lymphocytes and granulocytes were detected in extremely high titers. Immune-mediated pancytopenia was refractory on multiple immunosuppressive treatment strategies. Proliferation of polyclonal plasma cells of recipient-type that was documented postmortem, was most likely responsible for excessive antibody formation. PMID- 15136242 TI - Prostatic acid phosphatase: a possible diagnostic marker of intravascular large B cell lymphoma. AB - BACKGROUND AND OBJECTIVE: Intravascular large B-cell lymphoma (IVL) has been treated as fever of unknown origin (FUO), and many patients have been treated inadequately based on incorrect diagnoses. We previously cares for a patient with IVL who tested positive for prostatic acid phosphatase (PAP), a marker of prostate cancer. Since then, we have regularly examined this mather when IVL was suspected to investigate the usefulness of PAP as a diagnostic marker for IVL. We retrospectively evaluated the usefulness of PAP as diagnostic marker of IVL. DESIGN AND METHODS: We reviewed the clinical courses of 5 patients with IVL (3 males, 2 females) in comparison with 23 controls with hematologic malignancies other than IVL. RESULTS: Serum levels of PAP were elevated in all 5 patients with IVL and 2 of the 23 controls. The difference was statistically significant using a chi-squared test (p=0.0002). The sensitivity and specificity of PAP were 100% and 91%, respectively, in the diagnosis of IVL. Its serum levels were closely associated with disease status. INTERPRETATION AND CONCLUSIONS: This study suggests that PAP might be a useful marker for the screening and assessment of disease activity and responses to the treatment of IVL. PMID- 15136243 TI - Acquired factor XIII deficiency due to an inhibitor: a case report and review of the literature. PMID- 15136244 TI - Complete molecular remission in a patient with Philadelphia-chromosome positive acute myeloid leukemia after conventional therapy and imatinib. AB - Philadelphia-chromosome (Ph)-positive acute myeloid leukemia (AML) is rare and prognosis is poor with a median survival of six to seven months only. We report on a patient with Ph-positive AML (FAB M2, major BCR/ABL1 mRNA transcript, b2a2), who is in sustained complete cytogenetic and molecular remission for 15 months. Cytarabine-based chemotherapy was discontinued after two courses due to infectious complications. Since the b2a2 transcript was still detectable, imatinib was started with quantitative RT-PCR monitoring. This result is promising and worth further evaluation to establish the role of imatinib in patients with Ph-positive AML. PMID- 15136245 TI - T-cell receptor gammadelta-large granular lymphocytic leukemia associated with an aberrant phenotype and TCR-Vbeta20 clonality. AB - Large granular lymphocytic (LGL) leukemia is a rare heterogenous disorder of mature lymphocytes with a characteristic morphology, multiple autoimmune disorders and indolent clinical course. Most cases exhibit a T-cell phenotype of CD3, CD8 and CD57 positivity, while the minority exhibit a CD2, CD56, and CD16 positive NK-cell phenotype. We report a case of a 71-year-old female suffering from a TCRgammadelta positive T-cell leukemia with a morphology compatible to LGL leukemia. She referred to the hospital for investigation of mild anemia, lymphocytosis, neutropenia and hyperglobulinemia. Peripheral blood and bone marrow were occupied by mature large granular lymphocytes with abundant azurophilic granules. The immunophenotype was CD3+, CD2+, CD5+, CD7+, CD4-, CD8-, CD16-, CD56-, CD57- and the Vbeta repertoire analysis showed clonal reactivity with Vbeta20 mAb. The patient was diagnosed as having T-LGL and was treated with G-CSF. So far, she experiences an indolent clinical course. To our knowledge, this is a rare case of TCRgammadelta positive T-LGL leukemia with the aberrant immunophenotype of CD3+, CD4-, CD8-, CD16-, CD56-, CD57-. PMID- 15136246 TI - Bull's neck in Hodgkin's disease: a CT imaging appraisal. PMID- 15136247 TI - Atypical pyoderma gangrenosum in a patient with post-polycythaemic myelofibrosis. PMID- 15136248 TI - Granulocytic sarcoma of the lacrimal gland. PMID- 15136249 TI - Unrelated cord blood transplantation for children with high risk myelodysplastic syndromes. PMID- 15136250 TI - Biomechanics of normal and pathological gait: implications for understanding human locomotor control. AB - The biomechanical (kinetic) analysis of human gait reveals the integrated and detailed motor patterns that are essential in pinpointing the abnormal patterns in pathological gait. In a similar manner, these motor patterns (moments, powers, and EMGs) can be used to identify synergies and to validate theories of CNS control. Based on kinetic and EMG patterns for a wide range of normal subjects and cadences, evidence is presented that both supports and negates the central pattern generator theory of locomotion. Adaptive motor patterns that are evident in peripheral gait pathologies reinforce a strong peripheral rather than a central control. Finally, a three-component subtask theory of human gait is presented and is supported by reference to the motor patterns seen in a normal gait. The identified subtasks are (a) support (against collapse during stance); (b) dynamic balance of the upper body, also during stance; and (c) feedforward control of the foot trajectory to achieve safe ground clearance and a gentle heel contact. PMID- 15136251 TI - Perturbations of human posture: influence of impulse modality on EMG responses and cerebral evoked potentials. AB - Leg muscle EMG responses and cerebral evoked potentials (CP), elicited by perturbations of stance while on a treadmill with split belts, were analyzed in order to study the relationship between compensatory leg muscle responses and afferent input to supraspinal centers. Various conditions of perturbation were used to establish the extent to which compensatory EMG responses and CPs show congruent behavior. Four different treadmill acceleration rates were applied in three different conditions (unilateral perturbation, directed forward or backward; bilateral perturbation, directed forward or backward; and opposing bilateral perturbation). EMG responses and CPs showed parallel increases in amplitude with increasing displacement velocity. The EMG responses showed distinct differences, predominantly in the response amplitude, between the different perturbation conditions, whereas the CPs were affected only to a minor degree. Tibialis anterior EMG responses were more closely related to the CP following forward perturbation than the corresponding gastrocnemius responses were to the CP following backward perturbation. We conclude that the EMG responses are more closely related than the CPs to displacement parameters and suggest that this is due to the further spinal processing of the afferent input needed to generate an appropriate EMG response. The closer relationship between the tibialis anterior response and CP may reflect a predominant central representation and control of tibialis anterior activation in the regulation of posture. The functional implications of these findings are discussed. PMID- 15136252 TI - Qualitative dynamics of disordered human locomotion: a preliminary investigation. AB - A qualitative approach to the evaluation of disordered locomotion is introduced within the framework of dynamical systems theory. Exemplar phase plane and angle angle plots of knee and ankle movements were constructed from limb trajectories of neurologically impaired individuals and qualitatively compared with similar plots, reflecting normal locomotion. In phase plane trajectories of normal locomotion, characteristics of spring-like dynamics dominated the loading and unloading phases whereas those of ballistic pendular dynamics were seen during swing. The overall squareness of the normal phase plane trajectories suggested precisely timed and narrowly focused controls. in contrast, phase plane records from hemiparetic subjects had markedly reduced segmental velocities, pronounced velocity reversals in both stance and swing, and a loss of overall squareness. Knee-ankle plots of normal locomotion revealed important features of intersegmental coordination such as coupled out-of-phase coordination in loading and unloading, a decoupled phase offset in early swing, and a kind of active partitioning in late swing in which one segment moved while the other remained constant. These intersegmental relations were absent or distorted in the hemiparetic angle-angle plots. It is suggested that this qualitative approach, together with electromyography and force dynamics, may allow the characterization of the movement disorders associated with given neuropathologies. PMID- 15136253 TI - Development of stepping patterns in human infants: a dynamical systems perspective. AB - Upright locomotion has been, perhaps, the most studied of all human motor behaviors. The acquisition of this motor milestone has been of major interest to developmentalists from the motor as well as the cognitive domains. Until recently, developmentalists have chosen a nonexperimental approach, opting instead to describe the motor behaviors that precede walking, assigning them precursor status, and attributing the sequence solely to maturation of the nervous system. In this paper, I will discuss evidence that suggests that we may better understand this and other aspects of motor development by viewing them from the theoretical perspective of nonlinear and complex dynamical systems. Focus will be on the availability of a coordinated and alternating stepping pattern throughout much of the first year of life. Factors that influence the expression of and variability in this movement pattern will also be discussed. In addition, I will address the need to understand the development of control of pattern formations as well as their coordination, and I will discuss one approach that may provide some insight; that is, to study the modulations and patterns of muscle and other forces underlying limb movements. PMID- 15136254 TI - A developmental study of the interlimb coordination in running and galloping. AB - Using a dynamical systems perspective on motor behavior, it was predicted that interlimb coordination of running and galloping would behave like coupled, nonlinear, limit-cycle oscillators, which show the properties of phase locking, entrainment, and structural stability. Female subjects ranging in age from 2.5 years to adult were filmed while running and galloping with and without a weight perturbation. Analysis of both temporal- and amplitude-phasing measures revealed that both gaits demonstrated oscillatory properties. Differences between gaits and across age were primarily a matter of degree. In general, children 4 years of age and below had slightly less table phasing patterns, and all age groups showed slightly less ability in the gallop, particularly with amplitude phasing. PMID- 15136255 TI - An analysis of air-stepping in normal infant vervet monkeys. AB - Limb movements during air-stepping were analyzed in three neonatal vervet monkeys over a three-week period. The movements had similar temporal organization both across animals and across time. For example, the duration of both the hind and the forelimb cycle equaled about 500 ms, with hind limb return strokes lasting much longer than the hind limb power strokes. Furthermore, there were clear indications of both intra- and interlimb coordination. Specifically, all the joints of a limb tended to flex and extend simultaneously, and contralateral and ipsilateral limb pairs had an average phase relationship of approximately 50% of cycle duration. Despite a qualitative similarity between limb movements during air-stepping in the neonates and overground locomotion in older animals, there were notable differences both in temporal relationships and joint displacement patterns. Finally, there appeared to be important similarities between air stepping in these monkeys and stepping in newborn humans. Most notably, both tended to disappear after a limited period. The implications of these similarities, as well as the overall results, are discussed in relation to the understanding of the development of locomotor behavior in human and nonhuman primates, using approaches based both upon the hard-wired and dynamic models. PMID- 15136256 TI - The role of propulsive muscles of the shoulder during quadrupedalism in vervet monkeys (Cercopithecus aethiops): implications for neural control of locomotion in primates. AB - In comparative anatomical studies of the shoulder, the humeral retractors are often grouped together as propulsive muscles, which are important in the propulsive stroke of the forelimb during quadrupedal locomotion. Electromyographic (EMG) analyses of these muscles in opossums, cats, and dogs in general have confirmed such conclusions. An EMG study of chimpanzee shoulder muscles during knuckle-walking found, however, that the humeral retractors are either inactive or perform a function unrelated to propulsion (Larson & Stern, 1987). This contrast in muscle recruitment patterns between chimpanzees and more "typical" mammalian quadrupeds was attributed to the derived morphology of the chimpanzee shoulder. The present study examines the activity patterns of the humeral retractors in the vervet monkey, a primate more closely resembling nonprimate mammals in its shoulder morphology. The results of this EMG analysis show that despite the significant differences in anatomy between chimpanzees and vervets, the two species display very similar muscle recruitment patterns during quadrupedalism, and there is evidence for this same pattern in other species of primates. These differences in muscle activity patterns between primates and nonprimate mammals may be related to changes in the neurological control of locomotion in primates due to the evolutionary development of manipulative abilities in the primate forelimb. PMID- 15136257 TI - Spinal cord circuits: are they mirrors of musculoskeletal mechanics? AB - Over the past decade, research at three different levels of sensorimotor control has revealed a degree of complexity that challenges traditional hypotheses regarding servocontrol of individual muscles: (a) The connectivity of spinal circuits is much more divergent and convergent than expected. (b) The normal and reflex-induced recruitment of individual muscles and compartments of muscles is more finely controlled than was noted previously. (c) The mechanical interactions among linked skeletal segments and their often multiarticular muscles are neither simple nor intuitively obvious. We have developed a mathematical model of the cat hind limb that permits us to examine the influence of individual muscles on posture and gait. We have used linear quadratic control theory to predict the optimal distribution of feedback from a hypothetical set of proprioceptors, given different assumptions about the behavioral goals of the animal. The changes in these predictions that result from changes in the structure and control objectives of the model may provide insights into the functions actually performed by the various circuits in the spinal cord. PMID- 15136258 TI - A comparative review of the primate motor system. AB - Primates have evolved separately from other mammals since the late Cretaceous, and during this time the two major extant primate groups, prosimians (lorises, lemurs, and tarsiers) and anthropoids (monkeys, apes, and humans) arose. Concurrently, structures within the central nervous system acquired primate characteristics. Not all of the uniquely primate features have been identified in the brain, but several are well known. The pyramidal system, the best studied motor system, shows a distinct primate pattern in its terminal connections in the spinal cord. Other descending systems are less well known, but primate specializations in the vestibular system and red nucleus have been observed. The primary and secondary motor cortices are topographically separated in primates, suggesting one basis for increased complexity. Given the size of the brain, structures in the basal ganglia are relatively enlarged in primates as compared with other mammals, whereas the cerebellum has the same relative size. PMID- 15136259 TI - Nonsomatic cerebullar circuits: a broader view of cerebellar involvements in locomotion. AB - The cerebellum not only receives extensive proprioceptive input from the periphery but also has bidirectional monosynaptic connections with the hypothalamus. This latter structure, in turn, is reciprocally interconnected with brain-stem visceral centers that also project to the cerebellum. Through these connections, information derived from somatic receptors that arrives at the cerebellum can directly influence visceral centers and circuits. It is suggested that the visceral responses seen during locomotor activity can be continuously monitored and partially regulated through such connections. PMID- 15136260 TI - Cues and control strategies in visually guided tracking. AB - Detailed quantitative models are required to investigate the neurological basis of motor behavior. Previous studies of visually guided manual tracking have either identified a variety of control signals (cues) for planning tracking movements or analyzed how a single cue is used (i.e., one-tracking strategy). A systematic, quantitative analysis of the effects and interactions of cues in terms of human manual-tracking performance is presented here together with measurements of concomitant eye movements. These measurements help define the routes by which information reaches the CNS, and the analysis elucidates how the control signals are processed and combined. The results quantify not only the large improvement in performance observed when the target waveform being tracked is predictable but also the extent to which this improvement depends on the availability of current information about target movements and positional error. Target information is shown to provide short-term prediction independent of the error signals used in on-line negative feedback control. PMID- 15136261 TI - Effects of average movement velocity on reaction time and spatiotemporal accuracy in single-aiming and rapid-timing movement tasks. AB - The effects of instructed movement speed were investigated in two experiments. First, rapid-timing and single-aiming movement tasks were compared. Unlike rapid timing, single aiming implies spatial accuracy. The aim of the first experiment was twofold: (a) to examine whether the requirement of accurate placement termination in single aiming affects the negative relationship between instructed average velocity and reaction time found in rapid timing, and (b) to test the speed-accuracy relationships predicted by the symmetric impulse variability model of these movement tasks. For this purpose, four average velocities (5, 24, 75, and 140 cm/s) were investigated in both types of movement tasks in a two-choice reaction task. The effects of average velocity on reaction time were similar in both single-aiming and rapid-timing tasks, and the predicted linear relationship between instructed average velocity and spatial accuracy was not found. The results suggest that the movement control mode, that is, open loop or closed loop, interferes with effects of instructed average velocity. The movement control mode explanation was confirmed in the second experiment with respect to the effect of paired velocities on reaction time. It is argued that the type of movement control mode must be considered in the interpretation of effects of instructed average velocity on reaction time and spatiotemporal measures. PMID- 15136262 TI - Form and exploration of mechanical stability limits in erect stance. AB - The stability limits of erect stance are described as a geometrical structure in a movement space. Mechanical properties and response latency, a neural property, are determining factors for the stability limits. Standing stability limits of adults and young children are compared, and a simple scheme is suggested by means of which infants can discover the stability limits as they learn to stand. The mechanics of different standing movements are discussed because their spatial temporal properties relate directly to their different stability limits. The combination of the stability limits for different movements gives a total set of stability limits, a different structure for adults than for children. PMID- 15136263 TI - The effects of practice on limb kinematics in a throwing task. AB - The effect of practice on limb kinematics in a dart-throwing task was examined to test three current hypotheses regarding limb control: trajectory formation; end point control; and coordinated joint-space control. Practice was given to both the relatively well-practiced dominant ant the relatively unpracticed nondominant limbs of 5 male subjects to permit analysis of the early phase of coordination acquisition. The nondominant limb demonstrated high absolute joint cross correlations with high variability throughout practice and consistency in the hand trajectory. The dominant limb exhibit a significant decrease in wrist-elbow and wrist-shoulder cross-correlations over practice while also maintaining a consistent hand trajectory and significantly higher scoring performance. The findings demonstrate that practice effects can be seen in both coordination mode and variability of various parameters of limb motion, but the changing relationship between the variables suggests that control cannot be ascribed to any one of the three hypotheses advanced. It is proposed that the observed invariance or variance in limb trajectories, end-point control, and coordinated joint angles are a reflection of more global parameters emerging from the flow field properties of the organism, environment, and task interaction. PMID- 15136264 TI - Visual fixation and postural sway in children. AB - Adults are able to use a visual target to reduce quiet-standing postural sway (Lee & Lishman, 1975). The present study was designed to determine whether children, under varying postural conditions, are also able to use a visual target to reduce postural sway. A second purpose was to determine the ability of children to visually fixate under different postural conditions. An inability to visually fixate may limit the usefulness of a visual target. The results indicate that, like adults, children are able to reduce sway in the presence of a visual target. Young children are less able than older children and adults to visually fixate. In addition, children show more spontaneous visual saccades in the no target condition than in the target condition and more saccades in the Romberg stance than in a feet-together stance. The fact that saccades decrease with increasing age, even in the seated, head-stabilized condition, precludes the possibility that increased instability of the young children is the only cause of increased number of saccades. PMID- 15136265 TI - Complex oscillations in a human motor system. AB - Experiments were performed to investigate the oscillations arising in a human motor system with delayed visual feedback. Eight subjects were instructed to maintain a constant finger position relative to a stationary baseline. The finger displacement was measured using a microdisplacement transducer connected to the index finger, and was displayed on an oscilloscope. Time delays between 40 and 1,500 ms were inserted in the visual feedback loop for 100 s. Results show that, as the time delays increase, irregular rhythms appear with short intermittent periods of regular oscillations. These regular low-frequency oscillations have an amplitude that increases with the time delays and a period that is consistently about 2 to 4 times the time delay. Fast Fourier Transforms (FFT) show a peak between 8 and 12 Hz, corresponding to physiological tremor in half the subjects. No systematic variations in the FFT for the 2 to 15 Hz range were observed as time delay increased. In the 0 to 2 Hz range, the FFT show a consistent increase in power with the time delay. These results indicate that, under the conditions of this experiment, tremor is not affected by time delays in the visuomotor system, and time delays in the visuomotor feedback loop give rise to complex oscillatory behaviors. PMID- 15136266 TI - Backward walking: a simple reversal of forward walking? AB - The purpose of this study was to determine whether backward walking represented a simple temporal reversal of forward walking and, hence, could be controlled by a reversed cycling of the same group of neurons. Electromyographic (EMG), joint angle, joint moment, and joint muscle power patterns were compared for forward and backward walking, in 6 subjects. The joint angle patterns with the time-base of the backward walking reversed were similar, with the exception of the ankle. The moment patterns were similar except for the knee, whereas the joint muscle powers were almost reversed-polarity images of each other. This suggests that somewhat similar muscle activation patterns could be used to produce both modes of locomotion, but the temporal cycling of muscle contraction would be reversed: Concentric muscle activity in forward walking would become eccentric activity in backward walking, and visa versa. The EMG results generally supported these findings. PMID- 15136267 TI - Timecourse of preshaping for functional responses to objects. AB - The present study explores the unfolding of hand shaping the context of functional responding to common objects. The principal issue addressed by this study is whether distinct, identifiable preshapes are evidenced in preparation for four distinct types of hand contact, including nonprehensile as well as prehensile hand shapes, and for small and large surfaces of contact. All four shapes considered here (poke, pinch, palm and clench) showed evidence of distinct preshaping, with similar timecourses in relation to the onset of reaching. The initial separation of the fingers, formation of first identifiable preshape feature, and completion of the preshape appeared at essentially the same time relative tot he onset of reaching for all four hand shapes. The hand shapes did differ, however, in the time between the completion of a stable preshape and contact with the object. The time for preshape-to-contact was greater for hand shapes with small surfaces of contact than for those with large surfaces and for prehensile shapes than for nonprehensile. These differences are attributed to the precision required for the ultimate object-contact response. PMID- 15136268 TI - A device for the computer processed analysis of video frames. AB - This article briefly describes a system of analyzing movement sequences extracted from video images. The system uses an IBM-PC or compatible computer, equipped with a CGA screen and a mouse, and a videotape recorder and monitor. The coordinates of the points selected from the images with the mouse are stored in an ASCII-format file by a computer program. The principle behind this system is that it projects video images on a computer screen through a one-way mirror while avoiding parallax effects. PMID- 15136269 TI - A note on the information-theoretic basis of Fitts' law. AB - Fitts' law is an information-theoretic view of human motor behavior developed from Shannon's Theorem, 17, a fundamental theorem of communications systems. Using data from Fitts' original experiments, we demonstrate that Fitts' choice of an equation that deviates slightly from the underlying principal is perhaps unfounded, and that the relationship is improved by using an exact adaptation of Shannon's equation. PMID- 15136270 TI - The step cycle organization of infant walkers. AB - To walk independently, the human infant must coordinate its multisegmented body to achieve both postural stability and forward mobility. In this experiment, we sought evidence of the form this coordination takes in infants who have been walking 3 to 10 months. Using the Philippson phases to represent the step cycle, we analyzed film records of infants and adults walking at speeds from 2 to 4 km/hr. Our results reveal a clear relationship between walking speed and the F and E(3) Philippson phases of the step cycle in both the young walkers and the adults. Indeed, across all four phases of the step cycle, the infants exhibited an organization that was almost identical to that of the mature walkers. PMID- 15136271 TI - Phase-dependent compensatory responses to perturbation applied during walking in humans. AB - The interaction between the peripheral and the central regulation of locomotion was studied by examining the dependency of the response to unexpected perturbation on the phase of the step cycle. The changes in the latency and magnitude of various muscle responses to electrical stimulation of the toe and applied unexpectedly at different phases of the locomotor cycle in humans are described. The results show that response to perturbation is gated and modulated in both ipsi- and contralateral limb muscles. These muscle responses, when present, were always excitatory in nature. They were not correlated with the normal locomotor activity, thus suggesting a more complex organization of the response. Except for one muscle in the contralateral limb, the latency of the other muscle responses did not vary across the step cycle. in response to the perturbation, the appropriate phase of the step cycle was shortened. The results from this study suggest that the perturbation applied elicits a phase independent, normal ipsilateral flexor response in the tibialis anterior muscle, while the gating and modulation of other ipsi- and contralateral muscles provide appropriate phase-dependent adaptive response to maintain postural stability and continue with the ongoing task of locomotion. PMID- 15136272 TI - Task-dependent compensatory responses to perturbations applied during rhythmic movements in humans. AB - Modulation of the responses to perturbation applied during different phases of three rhythmic movements in humans-running, cycling, and hopping-was studied. The perturbation was an electrical stimulus. The results showed gating and modulation of the responses in both ipsi- and contralateral limb muscles. The responses during running and cycling were only excitatory in nature, while during hopping an inhibitory response was observed. These responses were not correlated with the normal activity during the movement. The latency of the response in general was not altered for different stimulation phases. The alterations in the step cycle demonstrated overt behavioral changes due to the responses. There were differences between the responses observed during these movements and walking. In running, the major adaptation to perturbations appears to be in the contralateral side as seen in the changes in the step cycle. During cycling (except for one phase) and hopping, the same set of muscles was activated in response to perturbation. This represents a simplifying strategy in response organization. The dependency of the response on the task characteristics, postural stability requirement, and external constraints imposed on the subject is discussed. These studies provide insights into task-dependent strategies adopted by the nervous system to meet unexpected perturbation during rhythmic movements in humans. PMID- 15136273 TI - The influence of walking speed and prior practice on locomotor distance estimation. AB - In an attempt to reconcile discrepant findings in the literature (Elliott, 1986; Thomson, 1983), walking speed, prior practice, and walking delay were manipulated to determine if they had an impact on locomotor distance estimation. Contrary to suggestions by Thomson (1986), walking variability in the direction of locomotion depended only on a subject' initial distance from the targets. PMID- 15136274 TI - Asymmetric division of labor in human skilled bimanual action: the kinematic chain as a model. AB - This article presents a tentative theoretical framework for the study of asymmetry in the context of human bimanual action. It is emphasized that in man most skilled manual activities involve two hands playing different roles, a fact that has been often overlooked in the experimental study of human manual lateralization. As an alternative to the current concepts of manual preference and manual superiority-whose relevance is limited to the particular case of unimanual actions-the more general concept of lateral preference is proposed to denote preference for one of the two possible ways of assigning two roles to two hands. A simple model describing man's favored intermanual division of labor in the model are the following. 1) The two hands represent two motors, that is, decomplexity is ignored in the suggested approach. 2) In man, the two manual motors cooperate with one another as if they were assembled in series, thereby forming a kinematic chain: In a right-hander allowed to follow his or her lateral preferences, motion produced by the right hand tends to articulate with motion produced by the left. It is suggested that the kinematic chain model may help in understanding the adaptive advantage of human manual specialization. PMID- 15136275 TI - A note on the kinematcis of drawing movements in children. AB - The relationship between instantaneous tangential velocity and radius of curvature (power 1/3) that is characteristic of adult subjects was found to be already present in children 3 to 11 years old. The correlation coefficient between these two parameters increases gradually with age without apparent discontinuities. However, even at the upper limit of the age range examined, the values are still lower than in the adults. PMID- 15136276 TI - Oscillator mechanisms in the human motor system: investigating their properties using the aftercontraction effect. AB - It is proposed that the human motor system is organized to use hardware and/or software non-linear oscillator mechanisms, the output of these oscillators being responsible for driving the limbs via signals to muscle groups. Following earlier theoretical development, it is argued that these muscle groupings act as a unit and themselves are likely to behave as a non-linear system. The attributes of non linear oscillators are many, and they are potentially significant for the explanation of motor behavior. This paper reviews and presents recent experiments that investigated the properties of muscular aftercontraction. The basic finding shows that subsequent to a period of moderate strain against a fixed surface the treated limb exhibits prolonged involuntary molar oscillations in the plane of the treatment. These results provide for the presence of driving oscillator mechanisms in the human motor apparatus. The mechanisms show generality of action in that directed attention can lead to oscillation of untreated limbs. Overall, the experiments showed that the movements exhibited the mutual interaction, synchronization, and preservation of phase relationships that are fundamental properties of non-linear oscillators. the picture that emerges is that these mechanisms can drive involuntary movements that are richly patterned: like slow versions of voluntary movements. The aftercontraction phenomenon proves to be an excellent tool for research on the oscillatory substrate of human motor organization. PMID- 15136277 TI - Controlling velocity in rapid movements. AB - It has often been reported that subjects prefer to use a strategy in which they vary movement velocity and peak amplitude in a linear fashion. In this study, control of velocity and amplitude in rapid reciprocating movements of the interphalangeal joint of the thumb was investigated by examining movement trajectories and patterns of activity in the extensor pollicis longus (EPL) and flexor pollicis longus (FPL) muscles. In controlling either amplitude or peak flexion velocity without constraint, subjects always used a strategy in which peak extension velocity and peak flexion velocity had strong linear correlations with movement amplitude. When they were required to keep either amplitude or peak flexion velocity fixed their movements were still biased toward a strategy in which peak velocity and movement amplitude covaried. It is suggested that the preferred strategy is related to a basic principle of scaling the magnitude and duration of a velocity profile in order to achieve different movement amplitudes. PMID- 15136278 TI - Amending limb movements to mechanical perturbations under choice reaction time conditions. AB - This paper examines the amendment of limb movements to mechanical perturbations under choice reaction time conditions. The two experiments reported were designed to examine the question of whether amendment latencies increase under choice conditions using two different choice manipulations. In Experiment 1, choice was manipulated by varying the duration of a mechanical perturbation in a known direction, while in Experiment 2 the strategy was to present perturbations of similar duration but unknown direction. Subjects produced discrete timing responses (700 ms-70 degrees) during which perturbations were interjected on random trials. The results reported reflect the outcome accuracy, the kinematics of the responses and latency of response modifications as a function of the various choice manipulations. The data demonstrated no statistically significant increases in reaction time as a function of choice manipulations. Although latencies did not significantly vary, both experiments demonstrated a significant decrease in response accuracy for the choice conditions. Collectively, the results are discussed in relation to the discrepancies between previous proprioceptive choice reaction time studies, which appear to stem from inconsistencies in both experimental methods and data analysis procedures. PMID- 15136279 TI - Impulse characteristics in rapid movement: implications for impulse-variability models. AB - Three untested assumptions of the impulse-variability model were examined in two experiments utilizing rapid, uni-planar limb movements. Experiment 1 varied movement distance (A) and movement time (MT) in a rapid-timing paradigm where the subject moved a lever through a certain distance in a certain time. Experiment 2 varied A in a reversal response where the S made a rapid elbow flexion and extension in a given MT. Displacement recordings were made on every trial. KR (knowledge of results) about MT was given after every trial. The results can be summarized as follows: (a) As predicted by the model, variations in impulse size and velocity were directly related to the impulses size; (b) There was no correlation between the accelerative and decelerative impulse durations recorded during the reversal response supporting the notion that the impulses might be independent; (c) Negative correlations (-.20 to -.50) were demonstrated between peak acceleration and impulse duration for both experiments, counter to the predictions of the model; and (d) Counter to the predictions of the model, timing error (VEt) increased as A decreased for rapid-timing responses. When the correlational results are taken into account, the model has the capacity to account for curvilinear relationships between relative timing error and movement speed. Overall, the results suggest that the impulse variability model requires some restructuring before it can be considered a viable model for the control of rapid limb movements. PMID- 15136280 TI - Chronometric analysis of the posturo-kinetic programming of voluntary movement. AB - Posturo-kinetic programming was investigated in a simple reaction time paradigm. Standing subjects performed voluntary upper limb elevations under three conditions differing by the importance of their destabilizing effect on the initial balance. Three indexes were considered: the reaction time (RT) corresponding to delay between the response signal (RS) and the onset of the upper limb acceleration at wrist level (Aw); the motor latency (ML) corresponding to delay between RS and onset of the earlier of the two accelerations at shank level (As) which was previously shown to indicate the onset of postural adjustments; and the postural anticipation (PA) corresponding to the delay between the onset of the earliest As and the onset of Aw. The results showed that the RT varied in relation to parameters of the forthcoming movement. RT variations were essentially due to differences in the PA. It appeared that the ML depended on cognitive factors while the PA depended on biomechanical ones. it is concluded that ML corresponds to the "true" reaction time while PA covers the earliest part of the motor response. PMID- 15136281 TI - Absolute error revisited: an accuracy indicator in disguise. AB - When target accuracy is defined as the probability that an individual will respond to an accuracy task within a fixed distance around the target, then the composite error measures, E and AE, are shown to be fairly strong indicators of target accuracy in a relative sense. When AE and E are compared, AE is shown to be an even stronger accuracy indicator than E for most reasonable accuracy requirements. This, plus the fact that AE has certain desirable properties in ANOVA procedures, suggests that AE is a good, composite measure of target accuracy and should be analyzed first to determine if target accuracy differences exist. Subsequent analyses of bias and/or variability are then recommended. PMID- 15136282 TI - Motion variability--its definition, quantification, and origin. AB - The concept of motion variability is discussed and a normalized measure for its quantification is introduced. An example demonstrates that this new measure constitutes a global indicator of the current state of a motor learning process. The causes of motion variations are briefly discussed. They include initial perturbations of the skeletal, muscular, and neural systems as well as perturbations due to incremental changes, during motion execution, of external forces, muscular parameters (fatigue), afferent sensory inputs, and of the motor programs controlling the execution of the motion. PMID- 15136283 TI - Once more on the equilibrium-point hypothesis (lambda model) for motor control. AB - The equilibrium control hypothesis (lambda model) is considered with special reference to the following concepts: (a) the length-force invariant characteristic (IC) of the muscle together with central and reflex systems subserving its activity; (b) the tonic stretch reflex threshold (lambda) as an independent measure of central commands descending to alpha and gamma motoneurons; (c) the equilibrium point, defined in terms of lambda, IC and static load characteristics, which is associated with the notion that posture and movement are controlled by a single mechanism; and (d) the muscle activation area (a reformulation of the "size principle")--the area of kinematic and command variables in which a rank-ordered recruitment of motor units takes place. The model is used for the interpretation of various motor phenomena, particularly electromyographic patterns. The stretch reflex in the lambda model has no mechanism to follow-up a certain muscle length prescribed by central commands. Rather, its task is to bring the system to an equilibrium, load-dependent position. Another currently popular version defines the equilibrium point concept in terms of alpha motoneuron activity alone (the alpha model). Although the model imitates (as does the lambda model) spring-like properties of motor performance, it nevertheless is inconsistent with a substantial data base on intact motor control. An analysis of alpha models, including their treatment of motor performance in deafferented animals, reveals that they suffer from grave shortcomings. It is concluded that parameterization of the stretch reflex is a basis for intact motor control. Muscle deafferentation impairs this graceful mechanism though it does not remove the possibility of movement. PMID- 15136284 TI - Information processes in movement learning: capacity and structural interference effects. AB - A series of experiments examined motor learning as an information processing activity occurring within a working short-term memory system and where response produced feedback and knowledge of results (KR) are used to modify the action plan developed from previous attempts at the movement task. Use of interpolated activities in the KR delay interval allowed inferences to be made regarding the capacity and structural characteristics of these information processes. Results indicated no capacity limitations on the learning process but important structural effects were found. The results supported the idea that response produced feedback is relatively unimportant as a feedback variable early in learning. Rather, the use of KR at a relatively high level of movement planning appears to be the important information processing activity underlying learning. Finally, the results supported the view that these information processes are related to cognitive problem solving activities. PMID- 15136285 TI - Development of the motor memory trace and control program. AB - A theoretical explanation of the development of the memory trace constrained to fit into a computer analog requires a coded reference "image" of the intended motor act, a tentative control program (memory trace) based on mobilizing and coordinating various pre-established motor-control subroutines, and an afferent neural description of the act as actually performed. The afferent information is non-consciously compared with the reference image, and the resulting discrepancy used to modify the control program so as to reproduce the reference image more exactly on the next attempt. The discrepancy consists of the sum of two types of differences, between image and program (present during learning) and the error difference between program and performance. The trial-by-trial discrepancies cumulate in the memory trace, so that during both learning and postlearning practice the error component results in a within-individual odd-even trial correlation. This constitutes a mechanism for a remoteness effect in the inter trial correlation matrix generated in a group of individuals during post-learning as well as learning conditions. Fifty-one individuals performed 50 trials on a simple arm movement task; 35 of these (the learners) showed individual systematic improvement during the first 20 trials but no improvement during the last 24; 16 (the non-learners) failed to improve with practice. Separate correlational analyses (both within- and inter-individual) of the two groups resulted in a variety of patterns that were predicted a priori in the learners by the theory, as did the 49 learners of another sample of 51 who performed a more complicated arm movement task. Also, as predicted, the non-learners failed to produce organized correlation patterns. PMID- 15136286 TI - Adaptation level as an explanation of the peak shift in generalization with movement stimuli. AB - Two experiments examine the value of adaptation level theory as an explanation of the peak shift phenomenon in stimulus generalization using linear arm movements. The first experiment manipulates the differences in adaptation levels during training by varying the frequency of experience of the training stimulus. Significant shifts in the peaks of generalization gradients were subsequently identified. The peak shifts were in the direction predicted by adaptation level theory. The second experiment manipulates testing adaptation levels by biasing the generalization test movements. Three groups of subjects performed differentially weighted test sequences in which the relative number of short, medium, or long movements was manipulated. Significant peak shifts of generalization gradients were observed in the direction predicted by adaptation level theory. PMID- 15136287 TI - Comorbid type 2 diabetes mellitus and hypertension exacerbates cognitive decline: evidence from a longitudinal study. AB - BACKGROUND: diabetes and hypertension are two highly prevalent diseases in the old population. They are highly related such that comorbidity is common. OBJECTIVES: to examine (i) the independent impact of the respective diseases on cognitive decline in very old age and (ii) the interactive impact of the two diseases on cognitive decline. SUBJECTS: 258 individuals (mean age = 83 years), all non-demented at baseline. Of these, 128 individuals (non-cases) were free from diabetes and hypertension, 92 individuals had a diagnosis of hypertension, 16 had a type 2 diabetes mellitus diagnosis without hypertension, and 22 had comorbid diabetes and hypertension. METHOD: a population-based longitudinal study of ageing (The OCTO-Twin Study), including four measurement occasions 2 years apart. The Mini-Mental State Examination was used to measure general cognitive function. Data were analysed using SAS Proc Mixed multilevel modelling. RESULTS: longitudinal trajectories indicated a steeper decline in cognitive function related to diabetes but not related to hypertension. However, the results indicated greatest cognitive decline among persons with comorbid diabetes and hypertension. CONCLUSIONS: it is concluded that comorbidity of diabetes and hypertension produce a pronounced cognitive decline. This finding emphasises the importance of prevention and treatment of those highly prevalent diseases in the old population. PMID- 15136288 TI - A randomised controlled trial of senior Lay Health Mentoring in older people with ischaemic heart disease: The Braveheart Project. AB - OBJECTIVE: to examine the effects and feasibility of educating and empowering older people with ischaemic heart disease using trained senior lay health mentors. DESIGN: randomised controlled trial with blinded evaluation. SETTING: Falkirk and District Royal Infirmary. PARTICIPANTS: inpatients and outpatients aged 60 or over attending secondary care with a diagnosis of angina or acute myocardial infarction. Three-hundred and nineteen entered and 289 completed exit assessments. The intervention group took part in mentoring groups for 1 year, meeting monthly for 2 hours, each led by two trained lay health mentors in addition to standard care. MAIN OUTCOME MEASURES: primary outcome measures were changes in coronary risk factors, medication usage and actual use of secondary care health services. Secondary outcomes were total and cardiovascular events; changes in medication compliance, non-medical support requirement, health status and psychological functioning, and social inclusion. RESULTS: there were significant improvements in a reported current exercise score (mean +0.33, +0.02 to +0.52), in the average time spent walking per week by 72 minutes (+1 to +137 minutes), and in the SF36 Physical Functioning Score (+6.1, +2.4 to +9.5). There was a 1.0% reduction in total fat (95% CI -3.0% to -0.6%) and a 0.6% reduction in saturated fat (95% CI -1.5% to -0.03%). The intervention group showed reduced outpatient attendance for coronary heart disease (-0.25 appointments, -0.61 to 0.08). Attendance rates were high. Socio-economic grouping did not affect participation. CONCLUSIONS: Lay Health Mentoring is feasible, practical and inclusive, positively influencing diet, physical activity, and health resource utilisation in older subjects with ischaemic heart disease without causing harm. PMID- 15136289 TI - Resolution of macroprolactinoma-induced symptomatic hydrocephalus following cabergoline therapy. AB - A 71-year-old man was referred because of memory loss. Magnetic resonance imaging showed a pituitary macroadenoma associated with hydrocephalus. Marked hyperprolactinaemia was present. After 2 months of cabergoline therapy, magnetic resonance imaging showed tumour shrinkage with resolution of the hydrocephalus. We report, for the first time, the adequate and rapid clinical response of a macroprolactinoma-induced symptomatic hydrocephalus in an elderly man to a low and once-a-week dose of cabergoline therapy. Medical therapy with this dopamine agonist in this particular patient was so effective that ventriculo-peritoneal shunting could be avoided. PMID- 15136290 TI - Geriatric medicine in Sweden: a study of the organisation, staffing and care production in 2000-2001. AB - OBJECTIVE: the organisation of long-term medicine and geriatric medicine has undergone many changes during the last 15 years. The aim of this study is to gain an overall perspective of the present organisation of geriatric medicine in Sweden. DESIGN: questionnaire survey. METHODS: The Swedish Society for Geriatric Medicine and Gerontology, in collaboration with the Federation of County Councils and the Swedish Association of Local Authorities, sent out a survey to people in all county councils in Sweden. The subject of the survey was the speciality of geriatric medicine in the Swedish healthcare system, with regard to healthcare organisation, staffing and care production in 2000/2001. RESULTS: there were 52 separate geriatric units, 41 independent 'clinics' and 11 'sections' within other departments. There were a total of 3,101 geriatric inpatient beds. On average, there was one geriatric bed for every 799 individuals within the local population aged 65 years and over, with a 10-fold variation between counties. Four counties had no geriatric provision. The 'geriatric clinics' were mainly located in university towns and averaged 85 beds per clinic, again with a 10-fold variation. There were 604 established positions for doctors within geriatrics, of which 63% were at geriatric clinics. On average, the clinics had 16 positions each (of which 75% were filled with geriatric specialists) with 7 beds per doctor. The corresponding averages for nurses and paramedics could not be summarised due to organisational differences between the county councils. In general, there were very few nurses with specialist training in geriatric medicine. CONCLUSIONS: the field of geriatric medicine in Sweden is very heterogeneous regarding terminology, designations, structure, staffing and care production. There is no overall structural plan for the role of geriatric medicine in Swedish healthcare, with the desired close connection between content and dimensioning of geriatric specialist training and the practical organisation of the activities. The county councils designate geriatric medicine so differently that it is hardly possible to compare different geriatric facilities today. Considering how many patients at hospitals today are elderly and suffer from multiple illnesses, it is a major quality issue to ensure that these patients have access to geriatric specialists. PMID- 15136292 TI - Bridging the chasm between bench and bedside: translational research in acute lung injury. PMID- 15136293 TI - Role of autoimmunity in organ allograft rejection: a focus on immunity to type V collagen in the pathogenesis of lung transplant rejection. AB - Lung transplantation is the only definitive treatment modality for many forms of end-stage lung disease. However, the lung is rejected more often than any other type of solid organ allograft due to chronic rejection known as bronchiolitis obliterans (BO). Indeed, BO is the primary reason why the 5- and 7-yr survival rates are worse for the lung than for any other transplanted organ. Alloimmunity to donor antigens is established as the primary mechanism that mediates rejection responses. However, newer immunosuppressive regimens designed to abrogate alloimmune activation have not improved survival. Therefore, these data suggest that other antigens, unrelated to donor transplantation antigens, are involved in rejection. Utilizing human and rodent studies of lung transplantation, our laboratory has documented that a native collagen, type V collagen [col(V)], is a target of the rejection response. Col(V) is highly conserved; therefore, these data indicate that transplant rejection involves both alloimmune and autoimmune responses. The role of col(V) in lung transplant rejection is described in this review article. In addition, the potential role of regulatory T cells that are crucial to modulating autoimmunity and alloimmunity is also discussed. PMID- 15136294 TI - p120: the guardian of endothelial junctional integrity. PMID- 15136295 TI - Matrix GLA protein modulates branching morphogenesis in fetal rat lung. AB - The regulation of matrix gamma-carboxyglutamic acid protein (MGP) expression during the process of lung branching morphogenesis and development was investigated. MGP mRNA expression was determined over an embryonic and postnatal time course and shown to be developmentally regulated. Immunohistochemical analysis revealed increased staining for MGP in peripheral mesenchyme surrounding distal epithelial tubules. Fetal lung explants were used as an in vitro growth model to examine expression and regulation of MGP during branching morphogenesis. MGP mRNA expression over the culture interval mimicked the in vivo time course. Explants cultured in the presence of antibodies against MGP showed gross dilation and reduced terminal lung bud counts, accompanied by changes in MGP, sonic hedgehog, and patched mRNA expression. Similarly, antifibronectin antibody treatment resulted in explant dilation and reduced MGP expression, providing evidence for an interaction with MGP and fibronectin. Conversely, intraluminal microinjection of anti-MGP antibodies had no effect either on explant growth or MGP expression, supporting the hypothesis that MGP exerts its effects through the mesenchyme. Taken together, the results suggest that MGP plays a role in lung growth and development, likely via temporally and spatially specific interactions with other branching morphogenesis-related proteins to influence growth processes. PMID- 15136296 TI - Adenovirus-mediated transfer of the 1-cys peroxiredoxin gene to mouse lung protects against hyperoxic injury. AB - 1-Cys peroxiredoxin (1-cysPrx) is a novel antioxidant enzyme that has been shown to reduce a broad spectrum of peroxides including phospholipid hydroperoxides. We tested the hypothesis that adenovirus-mediated transfer of the 1-cysPrx gene can protect lungs of mice from oxidant injury. Mice infected with AdLacZ/AdNull were used as a control (AdCon). X-galactosidase staining revealed widespread expression of the LacZ gene in airways and lung alveoli. Compared with AdCon, 1 cysPrx expression was increased about twofold at 3 days after adenovirus infection. Mice with increased Prx expression showed less loss of body weight and longer survival during exposure to 100% O(2) or to 85% O(2) for 4 days followed by 100% O(2). At 72 h of 100% O(2) exposure, AdPrx infection protected mouse lungs from injury as indicated by less pleural effusion, lower lung wet/dry weight, less protein and fewer nucleated cells in bronchoalveolar lavage fluid, and lower content of thiobarbituric acid-reactive substances and protein carbonyls in lung homogenate. These findings show that increased expression of 1 cysPrx through adenovirus-mediated gene transfer protects mouse lungs from hyperoxic injury and delays death. PMID- 15136297 TI - Role of macrophages in virus-induced airway hyperresponsiveness and neuronal M2 muscarinic receptor dysfunction. AB - Viral infections exacerbate asthma. One of the pathways by which viruses trigger bronchoconstriction and hyperresponsiveness is by causing dysfunction of inhibitory M(2) muscarinic receptors on the airway parasympathetic nerves. These receptors normally limit acetylcholine (ACh) release from the parasympathetic nerves. Loss of M(2) receptor function increases ACh release, thereby increasing vagally mediated bronchoconstriction. Because viral infection causes an influx of macrophages into the lungs, we tested the role of macrophages in virus-induced airway hyperresponsiveness and M(2) receptor dysfunction. Guinea pigs infected with parainfluenza virus were hyperresponsive to electrical stimulation of the vagus nerves but not to intravenous ACh, indicating that hyperresponsiveness was due to increased release of ACh from the nerves. In addition, the muscarinic agonist pilocarpine no longer inhibited vagally induced bronchoconstriction, indicating M(2) receptor dysfunction. Treating animals with liposome-encapsulated dichloromethylene-diphosphonate depleted macrophages as assessed histologically. In these animals, viral infection did not cause airway hyperresponsiveness or M(2) receptor dysfunction. These data suggest that macrophages mediate virus induced M(2) receptor dysfunction and airway hyperresponsiveness. PMID- 15136299 TI - Trust: can we create the time? PMID- 15136298 TI - Regulation of LPS-mediated inflammation in vivo and in vitro by the thiol antioxidant Nacystelyn. AB - Increased levels of proinflammatory cytokines are present in bronchoalveolar lavage fluid in various lung diseases. Redox-sensitive transcription factors such as NF-kappaB regulate gene transcription for these cytokines. We therefore studied the effect of a new thiol antioxidant compound, Nacystelyn (NAL), on IL-8 regulation in a human macrophage-derived cell line (THP-1). LPS (10 microg/ml) increased IL-8 release compared with control levels. This LPS activation was inhibited by coincubation with NAL (1 and 5 mM). Pretreatment with cycloheximide or okadaic acid, protein synthesis, and serine/threonine phosphatase inhibitors, respectively, did not modify inhibition of IL-8 release caused by NAL. NF-kappaB and C/EBP DNA binding were increased after LPS treatment compared with control, an effect inhibited by cotreatment with NAL. Activator protein (AP)-1 DNA binding was unaffected. The enhanced neutrophil chemotaxis produced by conditioned media from LPS-treated cells was inhibited when cells were cotreated with NAL. The selectivity of NAL inhibition upon IL-8 expression was studied. LPS-treated THP-1 cells also had higher levels of TNF-alpha, transforming growth factor (TGF)-beta1 and -3, MIP-1alpha and -beta, and RANTES gene expression. However, only LPS induced IL-8 and TGF-beta1 expressions were inhibited by NAL. An anti inflammatory effect of NAL was confirmed in vivo as shown by a reduction in LPS induced neutrophil recruitment to the lungs following instillation of NAL into the lungs. Our studies demonstrate that NAL has anti-inflammatory properties in vitro and in vivo, may therefore have a therapeutic role in lung inflammation, and has the advantage over other antioxidant agents in that it may be administrated by inhalation. PMID- 15136300 TI - Women and heart disease: the role of diabetes and hyperglycemia. AB - Cardiovascular disease (CVD) is the primary cause of death in women, and women with type 2 diabetes mellitus are at greater risk of CVD compared with nondiabetic women. The increment in risk attributable to diabetes is greater in women than in men. The extent to which hyperglycemia contributes to heart disease risk has been examined in observational studies and clinical trials, although most included only men or did not analyze sex differences. The probable adverse influence of hyperglycemia is potentially mediated by impaired endothelial function, and/or by other mechanisms. Beyond high blood glucose level, a number of other common risk factors for CVD, including hypertension, dyslipidemia, and cigarette smoking, are seen in women with diabetes and require special attention. Presentation and diagnosis of CVD may differ between women and men, regardless of the presence of diabetes. Recognizing the potential for atypical presentation of CVD in women and the limitations of common diagnostic tools are important in preventing unnecessary delay in initiating proper treatment. Based on what we know today, treatment of CVD should be at least as aggressive in women-and especially in those with diabetes-as it is in men. Future trials should generate specific data on CVD in women, either by design of female-only studies or by subgroup analysis by sex. PMID- 15136301 TI - Prolonged QTc interval and risks of total and cardiovascular mortality and sudden death in the general population: a review and qualitative overview of the prospective cohort studies. AB - BACKGROUND: In certain subgroups of patients, prolongation of the QTc interval may increase total and cardiovascular mortality due to life-threatening ventricular arrhythmias and sudden death. Nonetheless, whether modest prolongation of the QTc interval in the general population has clinical importance remains unclear. METHODS: We conducted a literature search from 1990 forward to identify all published prospective cohort studies evaluating the association between prolonged QTc interval and risks of total and cardiovascular mortality as well as sudden death. We reviewed each of the studies individually and then conducted a qualitative overview. RESULTS: The 7 prospective cohort studies identified included 36 031 individuals. There were 2677 (8.7%) individuals with prolonged QTc interval, defined as 440 milliseconds or greater. Whereas 1 study reported no association between prolonged QTc interval and mortality (relative risk, 1.02; 95% confidence interval, 0.70-1.49), the other 6 reported inconsistent associations overall as well as across subgroups defined by various characteristics including age, sex, and comorbidities. The reported associations for both cardiovascular mortality and sudden death were also inconsistent. In the overview, the only consistent findings were for the subgroup of patients with prior cardiovascular disease, in which relative risks ranged from 1.1 to 3.8 for total mortality, from 1.2 to 8.0 for cardiovascular mortality, and from 1.0 to 2.1 for sudden death. Further, in individuals without prior cardiovascular disease, associations were either absent or greatly attenuated; specifically, relative risks ranged from 0.9 to 1.6 for total mortality, from 1.2 to 1.7 for cardiovascular mortality, and from 1.3 to 2.4 for sudden death. CONCLUSIONS: There was no consistent evidence for increased risks of total or cardiovascular mortality or of sudden death, except perhaps for patients with prior cardiovascular disease. In the general population, if QTc interval prolongation is associated with any increase in mortality, that risk is likely to be small and difficult to detect reliably. PMID- 15136302 TI - Patent foramen ovale in cryptogenic stroke: current understanding and management options. AB - There is increasing interest in the association between patent foramen ovale (PFO) and documented stroke of unknown cause, commonly referred to as cryptogenic stroke. We reviewed the literature and, on the basis of the available data, designed a diagnostic and treatment algorithm for patients with PFO and cryptogenic stroke. Patent foramen ovale is relatively common in the general population, but its prevalence is higher in patients with cryptogenic stroke. Importantly, paradoxical embolism through a PFO should be strongly considered in young patients with cryptogenic stroke. There is no consensus on the optimal management strategy, but treatment options include antiplatelet agents, warfarin sodium, percutaneous device closure, and surgical closure. High-risk features in the patient's history (ie, temporal association between Valsalva-inducing maneuvers and stroke, coexisting hypercoagulable state, recurrent strokes, and PFO with large opening, large right-to-left shunt, or right-to-left shunting at rest, and a coexisting atrial septal aneurysm) should prompt PFO closure. PMID- 15136303 TI - Changing the clinical management of hereditary hemochromatosis: translating screening and early case detection strategies into clinical practice. AB - BACKGROUND: This article describes the effect of an extensive physician educational program on detection and management of hereditary hemochromatosis (HH) before and after a hemochromatosis population screening study. METHODS: We measured the changes in clinical management by medical chart review for newly diagnosed cases before and after the educational program. The effect on detection of HH cases was determined by mail survey to primary care physicians in our health system. RESULTS: The median age at diagnosis of HH was 54 years before the study and 45 years after the study (P =.12). In the same period, among those with diagnosed hemochromatosis, the mean prestudy ferritin level changed from 1848 ng/mL to 606 ng/mL after the study (P =.03). The mean number of units removed by phlebotomy to complete "de-ironing" in diagnosed patients was 40 U before the study and 18 U after the study (P =.06); the number of months required for de ironing was 15 months before the study and 6 months after the study (P =.02). Before the study, no primary care physician was screening for HH. Two years after completion of the study, 11% (6/54) of primary care physicians indicated they had continued to screen all patients for hemochromatosis. CONCLUSIONS: Early detection and treatment of iron overload may be facilitated by educating health care providers and insurers about an "iron-avid" state, occurring in healthy patients who would benefit from periodic surveillance of their iron status. Continued education of physicians concerning diagnosis and treatment of HH aids progress toward increased early case detection. PMID- 15136304 TI - Risk factors for venous thromboembolism in hospitalized patients with acute medical illness: analysis of the MEDENOX Study. AB - BACKGROUND: There is limited information about risk factors for venous thromboembolism (VTE) in acutely ill hospitalized general medical patients. METHODS: An international, randomized, double-masked, placebo-controlled trial (MEDENOX) has previously been conducted in 1102 acutely ill, immobilized general medical patients and has shown the efficacy of using a low-molecular-weight heparin, enoxaparin sodium, in preventing thrombosis. We performed logistic regression analysis to evaluate the independent nature of different types of acute medical illness (heart failure, respiratory failure, infection, rheumatic disorder, and inflammatory bowel disease) and predefined factors (chronic heart and respiratory failure, age, previous VTE, and cancer) as risk factors for VTE. RESULTS: The primary univariate analysis showed that the presence of an acute infectious disease, age older than 75 years, cancer, and a history of VTE were statistically significantly associated with an increased VTE risk. Multiple logistic regression analysis indicated that these factors were independently associated with VTE. CONCLUSIONS: Several independent risk factors for VTE were identified. These findings allow recognition of individuals at increased risk of VTE and will contribute to the formulation of an evidence-based risk assessment model for thromboprophylaxis in hospitalized general medical patients. PMID- 15136305 TI - The prevalence of reduced glomerular filtration rate in older hypertensive patients and its association with cardiovascular disease: a report from the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial. AB - BACKGROUND: The prevalence of reduced glomerular filtration rate (GFR) in older hypertensive patients and the relationship between level of GFR and cardiovascular disease (CVD) and its risk factors are not well known. METHODS: We evaluated baseline renal function in 40 514 hypertensive patients 55 years or older who were enrolled in the Antihypertensive and Lipid-Lowering Treatment to Prevent Heart Attack Trial (ALLHAT). We used the simplified Modification of Diet in Renal Disease study equation to estimate GFR and examined the prevalence of CVD in patients with different levels of GFR. RESULTS: Fifty-seven percent of patients had mild (60-89 mL/min per 1.73 m(2)), 17.2% had moderate (30-59 mL/min per 1.73 m(2)), and 0.6% had severe (0.21 mg/mL) associated with depression in 6914 noninstitutionalized men and women (age, 18-39 years) from the Third National Health and Nutrition Examination Survey (NHANES III). RESULTS: The prevalence of lifetime major depression was 5.7% for men and 11.7% for women. The prevalence of elevated CRP level was 13.7% for men and 27.3% for women. A history of major depression was associated with elevated CRP level (odds ratio [OR], 1.64; 95% confidence interval [CI], 1.20-2.24). The association between depression and CRP was much stronger among men than among women. Results were adjusted for age, African American race, body mass index, total cholesterol, log triglycerides, diabetes, systolic blood pressure, smoking status, alcohol use, estrogen use in women, aspirin use, ibuprofen use, and self-reported health status. Compared with men without a history of depression, CRP levels were higher among men who had a more recent (within 1 year) episode of depression (adjusted OR, 3.00; 95% CI, 1.39-6.48) and who had recurrent (>or=2 episodes) depression (adjusted OR, 3.55; 95% CI, 1.55-8.14). CONCLUSION: Major depression is strongly associated with increased levels of CRP among men and could help explain the increased risk of cardiovascular disease associated with depression in men. PMID- 15136312 TI - Patient characteristics and experiences associated with trust in specialist physicians. AB - BACKGROUND: Nearly half of all medical visits are to specialist physicians, yet little is known about patients' outpatient experiences with specialists or how patients' characteristics and experiences are related to trust in specialist physicians. METHODS: We surveyed patients who had a new patient visit with a cardiologist, neurologist, nephrologist, gastroenterologist, or rheumatologist practicing in hospital-based practices (response rate, 73%; N = 417) and inquired about their experiences with care and trust in the specialist physician. We used multivariable models to assess associations of patients' characteristics and experiences with trust. RESULTS: Most patients reported good experiences, and 79% reported complete confidence and trust in the specialist. Black patients were less trusting than white patients (risk ratio [RR], 0.5; 95% confidence interval [CI], 0.2-0.8). Patients were more trusting if they reported that the consultant listened (RR, 1.8; 95% CI, 1.0-2.5), received as much information as they wanted (RR, 1.6; 95% CI, 1.1-1.9), were told what to do if problems or symptoms continued, got worse, or returned (RR, 1.4; 95% CI, 1.2-1.5), were involved in decisions as much as they wanted (RR, 1.5; 95% CI, 1.2-1.8), and spent as much time as they wanted with the specialist (RR, 1.8; 95% CI, 1.3-2.2). CONCLUSIONS: Patients reported high levels of trust in specialist physicians after an initial visit. Several specific experiences were associated with higher trust, suggesting that efforts to improve patient-physician interactions may be successful at achieving trust. Such efforts should especially aim to optimize physicians' interactions with black patients, who were less trusting of specialist physicians. PMID- 15136313 TI - The diagnostic yield of a standardized approach to idiopathic sensory-predominant neuropathy. AB - BACKGROUND: Peripheral neuropathy is a common problem that often prompts a lengthy and expensive diagnostic evaluation. A rational, evidence-based diagnostic approach to peripheral neuropathy is desirable. Prior studies have focused on all patients presenting to a tertiary referral center with a diagnosis of unclassified neuropathy. However, most patients with peripheral neuropathy have primarily sensory symptoms. This study focuses on patients with sensory predominant neuropathy. The goal was to develop a focused diagnostic algorithm that can be easily applied in a general medical setting. METHODS: Patients referred with predominantly sensory symptoms and no previously defined cause were included and evaluated using a standard diagnostic approach. RESULTS: Among 138 patients, 25% had at least 1 first-degree relative with symptoms suggestive of neuropathy. Among laboratory studies, a 2-hour oral glucose tolerance test had the highest diagnostic yield (61%) and was more sensitive than other measures of glucose metabolism. Vitamin B(12) deficiency was identified in 2 patients. Results of serum protein electrophoresis, immunofixation, and antinuclear antibody testing were abnormal in less than 5% of patients, and these rates are similar to those found in the general population. Using this approach, only 31% of patients completing the recommended evaluation were found to have an idiopathic neuropathy. CONCLUSIONS: Patients with sensory-predominant neuropathy should be tested for glucose tolerance and vitamin B(12) concentration. The significance of abnormalities of serum protein electrophoresis and antinuclear antibodies is uncertain. Other tests should be performed only when the clinical scenario is suggestive. Patients with atypical features may benefit from referral to a peripheral neuropathy center. PMID- 15136314 TI - The safety of telephone management of presumed cystitis in women. PMID- 15136315 TI - Slitlamp, specular, and light microscopic findings of human donor corneas after laser-assisted in situ keratomileusis. AB - OBJECTIVE: To examine slitlamp, specular, and light microscopic features of human donor corneas known to have undergone laser-assisted in situ keratomileusis (LASIK). METHODS: Twenty-six donor corneas known to have undergone LASIK prospectively underwent slitlamp examination with particular attention to the presence of a flap edge, as well as specular microscopy with particular attention to the presence of highly reflective particles in the stroma corresponding to the LASIK interface. Central endothelial cell density and pachymetery were obtained. They were compared with 26 control donor corneas without LASIK. Eleven LASIK donor corneas were processed for histology. Twenty-six donor corneas with no known prior keratorefractive surgery also underwent similar slitlamp examination and specular microscopy to serve as controls. RESULTS: Twelve (46%) of 26 LASIK donor corneas had an obvious flap edge, and 10 (39%) had a subtle flap edge by slitlamp examination. Four (15%) had infiltrates by slitlamp examination, of which 3 were confirmed by histopathologic examination. Highly reflective particles were seen by specular microscopy in the stroma of 23 (88%) of 26 LASIK donor corneas, but only 1 (4%) of 26 control donor corneas had a single highly reflective particle in the stroma (P<.001). The mean central endothelial cell counts were similar: 2138 cells/mm(2) in the LASIK group compared with 2250 cells/mm(2) in the controls (P =.39). Vacuolization and pyknosis of keratocytes was a consistent histopathologic finding after LASIK. Metallic particles at the interface were not detected by histology. CONCLUSIONS: Detection of a flap edge by slitlamp examination may detect at least half of the donor corneas that may have undergone LASIK. The detection of highly reflective stromal particles may form an effective basis for screening for LASIK donor corneas using specular microscopy and requires further study. PMID- 15136316 TI - Changes in astigmatism after congenital cataract surgery and intraocular lens implantation: a comparative study. AB - OBJECTIVE: To evaluate the postoperative changes in astigmatism in the pseudophakic eyes of children who underwent 1 of 3 different types of surgical incisions for congenital cataract extraction with intraocular lens implantation, and in whom astigmatism of at least 3 diopters (D) was recorded 1 week after the operation. METHODS: We retrospectively reviewed the medical records of all the children in our department who had undergone surgery for nontraumatic cataract between 1992 and 2001. Cataract surgery with intraocular lens implantation was performed using 1 of 3 types of surgical incisions: a limbal incision, a scleral tunnel, or a clear corneal incision allowing the use of a foldable intraocular lens. In 28 children (32 eyes) aged 2 months to 11 years (mean +/- SD, 4.7 +/- 3.4 years), astigmatism of 3 D or more was found when assessed 1 week after surgery. The refraction was measured and recorded again 3 months and 5 months after surgery. The paired t test was used to compare the outcome variables. MAIN OUTCOME MEASURES: Refractive error 1 week, 3 months, and 5 months after surgery. RESULTS: Mean +/- SD astigmatism 1 week postoperatively was 5.8 +/- 2.2 D, 5.1 +/ 2.1 D, and 4.0 +/- 1.3 D in groups 1, 2, and 3, respectively. Thereafter, the astigmatic component of the refractive error underwent a spontaneous decline, reaching mean +/- SD values of 0.9 +/- 1.0 D, 1.6 +/- 1.6 D, and 1.0 +/- 0.8 D, respectively, in the 3 groups 5 months after the operation. The difference between the mean values at 1 week and at 5 months in each group was statistically significant (P <.001 in group 1; P =.01 in group 2; and P<.001 in group 3). CONCLUSION: Children who underwent extraction of congenital cataract and intraocular lens implantation by different surgical techniques showed a significant spontaneous reduction in astigmatism postoperatively. PMID- 15136317 TI - Comparison of scanning laser polarimetry using variable corneal compensation and retinal nerve fiber layer photography for detection of glaucoma. AB - OBJECTIVE: To compare retinal nerve fiber layer (RNFL) measurements obtained with scanning laser polarimetry (SLP) using variable corneal polarization compensation with standard red-free photography for detection of RNFL damage in glaucoma. METHODS: This observational, cross-sectional study included 1 eye of each of 42 patients with open-angle glaucoma, 32 patients suspected of having glaucoma, and 40 healthy subjects. The RNFL measurements using SLP with variable corneal compensation were obtained within 3 months of red-free photographs. Two independent observers graded RNFL photographs using a standardized protocol. Superior and inferior hemiretinas were scored separately, and a global score was obtained by averaging scores from each hemiretina. MAIN OUTCOME MEASURES: The RNFL photography scores were compared with RNFL thickness measurements obtained with SLP. The receiver operating characteristic (ROC) curves were constructed to assess the abilities of the different methods to differentiate glaucoma patients from healthy subjects. RESULTS: The RNFL thickness decreased with increased RNFL damage as assessed by photographs in both hemiretinas (R(2) = 15%-47%). The area under the ROC curve for the best SLP parameter, Nerve Fiber Indicator, was significantly greater than the area under the ROC curve for the global RNFL photography score (0.91 vs 0.84, P =.03). CONCLUSIONS: A moderate correlation was found between RNFL thickness measurements obtained with SLP and RNFL scores from red-free photographs. Compared with semiquantitative RNFL photography scores, the best SLP parameter had a higher diagnostic accuracy to separate glaucoma patients from healthy subjects. PMID- 15136318 TI - Immediate postoperative use of a topical agent to prevent intraocular pressure elevation after pars plana vitrectomy with gas tamponade. AB - OBJECTIVE: To determine whether a single topical aqueous suppressant applied immediately after pars plana vitrectomy with long-acting gas tamponade prevents intraocular pressure (IOP) elevation. METHODS: Fifty patients who met the inclusion criteria and underwent pars plana vitrectomy with long-acting gas tamponade were randomized to receive a combination of timolol maleate and dorzolamide hydrochloride, long-acting timolol alone, dorzolamide alone, or placebo at the conclusion of surgery. The IOP was checked by a portable, handheld tonometer (Tono-Pen) at the conclusion of surgery and at 5 hours, 1 day, and 1 week after surgery. RESULTS: There were no significant differences in IOP among the groups at the conclusion of surgery. The IOP at 5 hours after surgery (27.0 vs 17.4 mm Hg; P<.001) and 1 day after surgery (26.1 vs 19.9 mm Hg; P =.01) showed a statistically significant difference between the placebo and timolol dorzolamide groups. The timolol-dorzolamide group showed greater IOP control than either the timolol alone or the dorzolamide alone groups at 5 hours (P =.04 for both). CONCLUSION: The use of a single topical aqueous suppressant (timolol dorzolamide) given after pars plana vitrectomy with long-acting gas tamponade effectively prevents significant postoperative IOP elevation at 5 hours and 1 day after surgery. PMID- 15136319 TI - Morphometric analysis of angiograms of exudative lesions in age-related macular degeneration. AB - OBJECTIVES: To analyze the morphometric composition of lesion components in exudative age-related macular degeneration and to study the relationships between individual lesion components and choroidal neovascularization (CNV) subtype, at 2 time points. METHODS: Morphometric analysis of 98 sets of angiograms separated by an interval of at least 3 weeks, with no treatment delivered in the intervening period between angiograms. Area measurements of individual lesion components were made from digitally captured angiograms. Choroidal neovascularizations were classified into subtypes based on the proportions of classic CNV. Fully corrected distance visual acuity measured on logMAR Early Treatment of Diabetic Retinopathy Study charts was available at baseline and at a subsequent visit in 78 subjects. Data were analyzed using parametric and nonparametric tests, linear regression, and McNemar test of equal proportions. RESULTS: Wholly and predominantly classic CNVs were significantly smaller at initial presentation than minimally classic or occult with no classic CNVs. Lesions containing blood and lipid were also significantly larger than lesions not exhibiting these features. Lesions containing any classic CNV expanded at a significantly greater rate than lesions without classic CNV. Approximately 40% of lesions categorized as wholly classic CNV converted to predominantly classic CNV between baseline and the next follow up visit. CONCLUSION: The presence of classic leakage in exudative age-related macular degeneration is the most important risk factor for rapid expansion of CNV. PMID- 15136321 TI - Practical approach to management of retinoblastoma. AB - OBJECTIVE: To present a simplified approach to management of retinoblastoma using basic clinical features. DESIGN: In a prospective, nonrandomized, single-center clinical trial, 158 eyes of 103 patients with retinoblastoma were managed with 6 cycles of chemoreduction (vincristine sulfate, etoposide, and carboplatin). The eyes were classified according to the Reese-Ellsworth classification and were also grouped on the basis of clinical features as follows: group 1, tumor only; group 2, tumor plus subretinal fluid; group 3, tumor plus focal seeds (3a, focal subretinal seeds; 3b, focal vitreous seeds); group 4, tumor plus diffuse seeds (4a, diffuse subretinal seeds; 4b, diffuse vitreous seeds); and group 5, neovascular glaucoma or invasive retinoblastoma. MAIN OUTCOME MEASURE: Treatment success (avoidance of enucleation and external beam radiotherapy). RESULTS: According to the Reese-Ellsworth classification, chemoreduction was successful in 100% of group Ia, 100% of group Ib, 86% of group IIa, 100% of group IIb, 91% of group IIIa, 100% of group IIIb, 50% of group IVa, 77% of group IVb, 50% of group Va, and 27% of group Vb. There was erratic correlation of the Reese-Ellsworth classification with treatment success. In contrast, the simplified grouping system displayed a smooth, nonerratic correlation for treatment success, with 100% success for group 1, 91% for group 2, 59% for group 3, and 12% for group 4 (group 5 always managed by primary enucleation). When all 6 subcategory groups were analyzed, there was consistent correlation for treatment success of 100% for group 1, 91% for group 2, 68% for group 3a, 54% for group 3b, 17% for group 4a, and 11% for group 4b. CONCLUSION: This practical approach to retinoblastoma using basic clinical features is predictive of treatment success for eyes in which modern conservative therapy for retinoblastoma is used. PMID- 15136320 TI - Associations of mortality with ocular disorders and an intervention of high-dose antioxidants and zinc in the Age-Related Eye Disease Study: AREDS Report No. 13. AB - OBJECTIVE: To assess the association of ocular disorders and high doses of antioxidants or zinc with mortality in the Age-Related Eye Disease Study (AREDS). METHODS: Baseline fundus and lens photographs were used to grade the macular and lens status of AREDS participants. Participants were randomly assigned to receive oral supplements of high-dose antioxidants, zinc, antioxidants plus zinc, or placebo. Risk of all-cause and cause-specific mortality was assessed using adjusted Cox proportional hazards models. RESULTS: During median follow-up of 6.5 years, 534 (11%) of 4753 AREDS participants died. In fully adjusted models, participants with advanced age-related macular degeneration (AMD) compared with participants with few, if any, drusen had increased mortality (relative risk [RR], 1.41; 95% confidence interval [CI], 1.08-1.86). Advanced AMD was associated with cardiovascular deaths. Compared with participants having good acuity in both eyes, those with visual acuity worse than 20/40 in 1 eye had increased mortality (RR, 1.36; 95% CI, 1.12-1.65). Nuclear opacity (RR, 1.40; 95% CI, 1.12-1.75) and cataract surgery (RR, 1.55; 95% CI, 1.18-2.05) were associated with increased all cause mortality and with cancer deaths. Participants randomly assigned to receive zinc had lower mortality than those not taking zinc (RR, 0.73; 95% CI, 0.61 0.89). CONCLUSIONS: The decreased survival of AREDS participants with AMD and cataract suggests that these conditions may reflect systemic rather than only local processes. The improved survival in individuals randomly assigned to receive zinc requires further study. PMID- 15136322 TI - Biocompatibility of trypan blue with human corneal cells. AB - OBJECTIVE: To quantify the toxicity of trypan blue on human corneal cells according to exposure time and concentration. METHODS: Three in vitro experiments were performed. (1) We exposed cultured human corneal fibroblasts to trypan blue (0.0001% to 0.1%) in Eagle modified minimum essential medium (EMEM) or phosphate buffered saline (PBS) for 15 minutes to 24 hours. Cytotoxicity was evaluated by Mosmann's colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MMT) assay. (2) We exposed human corneas in EMEM for 24 hours to trypan blue (0.001% to 0.1%). Fellow donor corneas served as controls. Endothelial survival was evaluated morphologically and by cell density assessment. (3) We morphologically compared the endothelial viability of human donor corneas after exposure to 0.1% trypan blue for 5 to 30 minutes with control corneas. RESULTS: In experiment 1, trypan blue in EMEM was not significantly toxic at concentrations of 0.005% or lower. Higher concentrations were toxic only after exposure to trypan blue for at least 6 hours. In PBS, significant toxicity was found after exposure to 0.1% trypan blue for 30 minutes or longer. Lower concentrations were toxic after longer exposures. In experiment 2, exposure to 0.01% and 0.1% trypan blue for 24 hours resulted in significant loss in cell density. At lower concentrations, the endothelium was affected only morphologically. In experiment 3, endothelial morphology changed in control corneas and after exposure to 0.1% trypan blue for as little as 5 minutes. After 30-minute exposure, morphologic deterioration was more pronounced. CONCLUSIONS: Trypan blue was toxic in vitro to corneal endothelium and corneal fibroblasts at higher concentrations and notably longer exposure times. Toxicity was less in EMEM than in PBS. Clinical Relevance At commonly used concentrations, both during cataract surgery and in the cornea bank, trypan blue is safe for corneal cells. At higher concentrations or longer exposures, however, caution is warranted. PMID- 15136323 TI - CD4-CD8 and CD28 expression in T cells infiltrating the vitreous fluid in patients with proliferative diabetic retinopathy: a flow cytometric analysis. AB - OBJECTIVES: To investigate CD4-CD8 and CD28 expression in T cells infiltrating the vitreous fluid in patients with proliferative diabetic retinopathy and to evaluate the relationship between the infiltrating T cells and both the activity of proliferative diabetic retinopathy and the clinical outcome. METHODS: Both vitreous and peripheral blood samples were obtained simultaneously from 20 consecutive diabetic patients and analyzed by flow cytometry. Three diabetic patients were excluded because there were no viable cells in the vitreous fluid. Six nondiabetic patients requiring vitrectomy were also studied. RESULTS: T lymphocytes were detected in all 6 diabetic patients with vitreous hemorrhage and in 6 (55%) of the 11 diabetic patients without vitreous hemorrhage, but in none of the nondiabetic patients. The percentages of T cells (CD3+), TCD4+ (CD3+ CD4+), and TCD8+ (CD3+ CD8+) subsets, as well as the expression of CD28, were similar in the vitreous fluid and in the peripheral blood in patients with vitreous hemorrhage. However, in patients without vitreous hemorrhage, the percentage of CD4+ CD28-T cells in the vitreous fluid was significantly higher than in the peripheral blood (33.34% [20.75%-100.00%] vs 8.45% [2.43%-56.59%]; P =.02). In addition, all of these patients showed quiescent retinopathy and their outcome was better than that of patients with vitreous hemorrhage and patients in whom intravitreous T cells were undetectable. CONCLUSION: T cells infiltrating the vitreous of diabetic patients without vitreous hemorrhage not only show a different pattern than in the peripheral blood but also seem to improve the prognosis of proliferative diabetic retinopathy. CLINICAL RELEVANCE: Our results provide further understanding of events involved in the autoimmune response in diabetic retinopathy and may aid in the research for new treatment approaches. PMID- 15136324 TI - Sunlight and the 10-year incidence of age-related maculopathy: the Beaver Dam Eye Study. AB - OBJECTIVE: To examine the association of sunlight exposure and indicators of sun sensitivity with the 10-year incidence of age-related maculopathy (ARM). DESIGN: Population-based cohort study. PARTICIPANTS: We included persons aged 43 to 86 years at the baseline examination from 1988 to 1990, living in Beaver Dam, Wis, of whom 3684 persons underwent 5-year follow-up and 2764 underwent 10-year follow up. METHODS: Data on sun exposure and indicators of sun sensitivity were obtained from a standardized questionnaire administered at baseline and/or follow-up. We determined ARM status by grading stereoscopic color fundus photographs using the Wisconsin Age-Related Maculopathy Grading System. MAIN OUTCOME MEASURES: Incidence and progression of ARM. RESULTS: While controlling for age and sex, we found that participants exposed to the summer sun for more than 5 hours a day during their teens, in their 30s, and at the baseline examination were at a higher risk of developing increased retinal pigment (risk ratio [RR], 2.99; 95% confidence interval [CI], 1.18-7.60; P =.02) and early ARM (RR, 2.20; 95% CI, 1.02-4.73; P =.04) [corrected] by 10 years than those exposed less than 2 hours per day during the same periods. In participants reporting the highest summer sun exposure levels in their teens and 30s, the use of hats and sunglasses at least half the time during the same periods was associated with a decreased risk of developing soft indistinct drusen (RR, 0.55; 95% CI, 0.33-0.90; P =.02) and retinal pigment epithelial depigmentation (RR, 0.51; 95% CI, 0.29-0.91; P =.02). Participants who experienced more than 10 severe sunburns during their youth were more likely than those who experienced 1 or no burn to develop drusen with a 250 microm diameter or larger (RR, 2.52; 95% CI, 1.29-4.94 [corrected] P =.01) by the 10-year examination. No relationships were found between UV-B exposure, winter leisure time spent outdoors, skin sun sensitivity, or number of bad sunburns experienced by the time of the baseline examination and the 10-year incidence and progression of ARM or its associated lesions. CONCLUSIONS: Few significant relationships between environmental exposure to light and the 10-year incidence and progression of ARM were found in the Beaver Dam Eye Study. Consistent with results from the baseline and 5-year follow-up examinations, significant associations were found between extended exposure to the summer sun and the 10 year incidence of early ARM and increased retinal pigment. A protective effect of hat and sunglasses use by participants while in their teens and 30s against the 10-year incidence of soft indistinct drusen and retinal pigment epithelial depigmentation was also found, but only in those who reported the highest amount of sun exposure during the same periods. PMID- 15136325 TI - Relative contributions of reduced vision and general health to NEI-VFQ scores in patients with neovascular age-related macular degeneration. AB - OBJECTIVE: To estimate the relative contributions of central vision loss and general health to vision-targeted quality of life as measured by the National Eye Institute Visual Function Questionnaire (NEI-VFQ). METHODS: Data on quality of life (NEI-VFQ and the 36-Item Short-Form Health Survey [SF-36]) and visual acuity were collected as part of the Submacular Surgery Trials Pilot Study. Information on medical conditions was collected by patient chart review. Twenty-four-month data for 120 patients were analyzed using linear regression methods. RESULTS: Median patient age at the 24-month examination was 77 years; 60% were women, and 98% were non-Hispanic whites. A 3-line decrement in visual acuity in the better seeing eye was associated with a 5.1- to 17.1-point decrement in NEI-VFQ scores after adjustment for general health (SF-36 physical component summary [PCS] and mental component summary [MCS] scores). A 10-point decrement in the PCS score was associated with a 4- to 9-point decrement in NEI-VFQ scores after adjustment for visual acuity in the better-seeing eye and MCS score. A 10-point decrement in the MCS score was associated with a 4- to 8-point decrement in NEI-VFQ scores after adjustment for visual acuity in the better-seeing eye and PCS score. Diabetes, arthritis/rheumatism, and hypertension also had large effects on NEI-VFQ scores in the adjusted analysis. CONCLUSIONS: The NEI-VFQ is sensitive to differences in visual acuity in the better-seeing eye, as expected, and to differences in general health. Adjustment for general health should be considered when comparing NEI-VFQ scores between patient groups. PMID- 15136326 TI - Maintenance of certification and the outside world. PMID- 15136327 TI - Industry funding for continuing medical education: is it ethical? PMID- 15136328 TI - Ethical concerns in industry support of continuing medical education: the con side. PMID- 15136329 TI - Pleomorphic adenoma of the lacrimal gland with extensive calcification. PMID- 15136330 TI - Bony hamartoma of the inferior orbital rim in a patient with tuberous sclerosis. PMID- 15136331 TI - Total opacification of intraocular lens implant after uncomplicated cataract surgery: a case series. PMID- 15136332 TI - Central serous chorioretinopathy after local application of glucocorticoids for skin disorders. PMID- 15136333 TI - Ocular involvement in systemic vasculitis associated with perinuclear antineutrophil cytoplasmic antibodies. PMID- 15136334 TI - Orbital osteoma with gaze-evoked amaurosis. PMID- 15136335 TI - Progressive pigmentation of chorioretinal lesions in aicardi syndrome. PMID- 15136336 TI - Lacrimal gland fistula. PMID- 15136337 TI - Ectopic lacrimal gland vs ectopic lacrimal ductule. PMID- 15136338 TI - Lacrimal gland secretion mimicking epiphora. PMID- 15136340 TI - Extension of retinal hemorrhage into the vitreous of a shaken baby through a break in the internal limiting membrane. PMID- 15136342 TI - Infliximab-associated retrobulbar optic neuritis. PMID- 15136344 TI - The Department of Surgery at the School of Medicine at the University Of Pittsburgh, Pittsburgh, Pa. PMID- 15136343 TI - Soya food intake and risk of endometrial cancer among Chinese women in Shanghai: population based case-control study. AB - OBJECTIVE: To evaluate the association of intake of soya food, a rich source of phytoestrogens, with the risk of endometrial cancer. DESIGN: Population based case-control study, with detailed information on usual soya food intake over the past five years collected by face to face interview using a food frequency questionnaire. SETTING: Urban Shanghai, China. PARTICIPANTS: 832 incident cases of endometrial cancer in women aged of 30 to 69 years diagnosed during 1997-2001 and identified from the Shanghai Cancer Registry; 846 control women frequency matched to cases on age and randomly selected from the Shanghai Residential Registry. MAIN OUTCOME MEASURES: Odds ratios for risk of endometrial cancer in women with different intakes of soya foods. RESULTS: Regular consumption of soya foods, measured as amount of either soya protein or soya isoflavones, was inversely associated with the risk of endometrial cancer. Compared with women with the lowest quarter of intake, the adjusted odds ratio of endometrial cancer was reduced from 0.93 to 0.85 and 0.67 with increasing quarter of soya protein intake (P for trend 0.01). A similar inverse association was observed for soya isoflavones and soya fibre intake. The inverse association seemed to be more pronounced among women with high body mass index and waist:hip ratio. CONCLUSION: Regular intake of soya foods is associated with a reduced risk of endometrial cancer. PMID- 15136345 TI - Education, economics, and excellence. PMID- 15136346 TI - Long-term detrimental effect of bile duct injury on health-related quality of life. AB - HYPOTHESIS: Long-term quality of life (QOL) in patients undergoing laparoscopic cholecystectomy (LC) incurring bile duct injury (BDI) and repair is comparable to that of patients undergoing uncomplicated LC. DESIGN: Case comparison study. SETTING: Secondary and tertiary care centers. PATIENTS: Eighty-six patients incurring BDI during LC between January 1, 1991, and July 31, 2003, were surveyed. Comparison subjects underwent uncomplicated LC during the same period. MAIN OUTCOME MEASURES: Health-related QOL as assessed by the Karnofsky Performance Scale, Medical Outcomes Study 36-Item Short Form Health Survey (SF 36), and Psychosocial Adjustment to Illness Scale. RESULTS: Fifty patients with BDI (39 [78%] female; mean +/- SEM age, 55 +/- 2 years) and 74 patients with uncomplicated LC (51 [69%] female, mean +/- SEM age, 52 +/- 2 years) responded. Of the 50 BDI patients, 48 (96%) had no stricture and normal liver function at QOL assessment. The mean +/- SEM follow-up period to QOL assessment for the BDI and uncomplicated LC groups was 62 +/- 6 and 47 +/- 3 months, respectively. The mean +/- SD Karnofsky Performance Scale scores were 77 +/- 9 vs 93 +/- 8 for the 2 groups, respectively (P <.001). The mean +/- SD SF-36 physical component scale scores after BDI vs uncomplicated LC were 36 +/- 11 vs 47 +/- 12, respectively (P <.001), compared with 50 +/- 10 for the normal population (P <.001). The mean +/- SD SF-36 mental component scale scores were 43 +/- 14 vs 49 +/- 11 for the 2 groups, respectively (P =.02), compared with 50 +/- 10 for the normal population (P =.01). Patients with BDI scored poorer on the Psychosocial Adjustment to Illness Scale health care orientation and domestic environment scales (P=.01). CONCLUSION: After BDI and repair, there are long-term detrimental effects of BDI on health-related QOL. PMID- 15136347 TI - Quality-of-life assessment of surgical reconstruction after laparoscopic cholecystectomy-induced bile duct injuries: what happens at 5 years and beyond? AB - HYPOTHESIS: Quality of life of patients after biliary reconstruction for laparoscopic injuries is comparable to that of patients after laparoscopic cholecystectomy. DESIGN: Outcomes study. SETTING: Tertiary referral center. PATIENTS: Records of 59 consecutive patients undergoing surgical reconstruction of the biliary tract after injury induced by laparoscopic cholecystectomy between 1990 and 1997 were reviewed. Hepp-Couinaud technique or Roux-en-Y hepaticojejunostomy was used in 53 patients; other procedures included cholangiojejunostomy, choledochorrhaphy, and hepaticoduodenostomy. INTERVENTIONS: Quality-of-life questionnaires (36-Item Short-Form Health Survey [SF-36]) were mailed to each patient in the group and to patients who underwent uneventful laparoscopic cholecystectomy, matched individually by year, sex, and age group. Values from the general population matched by age and sex were gathered (national norms). Minimum time of follow-up was 5 years. RESULTS: Eighty-nine (81%) of 110 potential respondents to the survey completed the SF-36 questionnaires. All 8 values evaluated in the SF-36 questionnaire (physical functioning, role-physical, bodily pain, general health perceptions, vitality, social functioning, role emotional, and mental health index) for patients undergoing biliary reconstruction were similar to those of both their matched controls (all P >.10) and national norms (all P >.05). The standardized physical component scale was also similar between the 2 groups (cases vs controls, 51 vs 48; P =.47), as was the standardized mental component scale (cases vs controls, 55 vs 55; P =.60). CONCLUSIONS: With a minimum of 5 years of follow-up, the quality of life after surgical biliary reconstruction compares favorably with that of both patients undergoing uneventful laparoscopic cholecystectomy and national norms. PMID- 15136348 TI - Resident work hours: what they are really doing. AB - HYPOTHESIS: We attempted to better quantitate resident work within our system of care. DESIGN: Survey. SETTING: Academic training program. PARTICIPANTS: Surgical residents. INTERVENTIONS: A work-hour survey was developed defining 5 areas of activity: patient care related to educational objectives, required educational activities, patient care activities unrelated to educational objectives, off-duty educational activity, and off-duty hours. MAIN OUTCOME MEASURES: Total work hours and noneducational work hours were analyzed by resident level, rotation, and category. RESULTS: The survey response rate was 52%, covering 110 workweeks. Residents worked 80 hours or less for 57 weeks and more than 80 hours for 53 weeks. The mean number of hours worked was 77. Fewer than one quarter (21.9%) of work hours were unrelated to educational activities. The amount of time spent in noneducational activities was lowest at community hospitals (17%) and similar at the Veterans Affairs (23%) and academic (22%) medical centers. It did not vary by total hours worked, averaging 21% for rotations of more than 80 h/wk and 23% for rotations of 80 h/wk or less. CONCLUSIONS: Residents spend a large amount of time in noneducational activities. Eliminating these activities would bring our rotations into compliance with the 80-hour workweek. It would also generate a large amount of time for educational activities within our training program. PMID- 15136349 TI - Advantages of focused helical computed tomographic scanning with rectal contrast only vs triple contrast in the diagnosis of clinically uncertain acute appendicitis: a prospective randomized study. AB - HYPOTHESIS: Focused helical computed tomographic (CT) scanning with rectal contrast only is a superior diagnostic modality compared with the traditional triple-contrast CT scan for the diagnosis of acute appendicitis. DESIGN: Prospective randomized analysis of both CT scan modalities. INTERVENTIONS: Only patients with uncertain diagnosis of acute appendicitis were entered in the study. The patients were then randomized to undergo the traditional triple contrast CT scan or the new focused CT scan with rectal contrast only. Surgical management included operation or observation for 23 hours. RESULTS: Ninety-one patients participated in the study, including 52 in the triple-contrast group and 39 in the rectal-contrast group. The demographics of the triple-contrast vs the rectal-contrast groups were similar. The triple-contrast group had a sensitivity of 97%, specificity of 86%, positive predictive value of 90%, and negative predictive value of 93%. The rectal-contrast group had a sensitivity of 88%, specificity of 100%, positive predictive value of 100%, and negative predictive value of 100%. There were 4 false-positive findings and 1 false-negative finding in the triple-contrast group compared with none in the rectal-contrast group. In the triple-contrast group, there were 13 perforated appendixes compared with 1 in the rectal-contrast group. The cost of a triple-contrast scan was 620 US dollars compared with 305 US dollars for a focused rectal-contrast scan. The negative appendectomy rate for the study was 8.0% (4 of 48 patients in the triple-contrast group vs 3 of 39 in the rectal-contrast group). CONCLUSIONS: The demographics, sensitivity, specificity, and positive and negative predictive values were comparable in both groups. The focused rectal-contrast procedure was better tolerated by patients and demonstrated decreased morbidity, delay to diagnosis, perforation rate, and negative appendectomy rate with no missed diagnosis and decreased cost. Therefore, we believe that focused helical CT scanning with rectal contrast only is a superior diagnostic modality compared with the traditional triple-contrast CT scan for the diagnosis of acute appendicitis. PMID- 15136350 TI - An algorithm to maximize use of minimally invasive parathyroidectomy. AB - HYPOTHESIS: Minimally invasive parathyroidectomy (MIP) depends on accurate preoperative localization of abnormal parathyroid glands. If the findings of a technetium Tc 99m sestamibi-labeled single-photon emission computed tomography (SPECT) (hereafter referred to as sestamibi SPECT or scan) are negative or ambiguous, cervical ultrasonography (CUS) may increase the success of preoperative gland localization and MIP, avoiding bilateral neck exploration. DESIGN: We collected data regarding preoperative sestamibi SPECT and CUS for parathyroid gland localization and intraoperative findings. SETTING: Tertiary care university hospital. PATIENTS: From August 1, 2000, through January 31, 2003, 71 patients (12 men and 59 women; mean age, 59 years) with primary hyperparathyroidism underwent preoperative sestamibi SPECT and CUS. Patients with prior or concurrent thyroid surgery, reoperative parathyroid disease, secondary/tertiary hyperparathyroidism, or studies performed at outside hospitals, were excluded. The MIP was performed by 1 surgeon with a 2- to 3-cm incision made on the side of the neck where the abnormal gland was preoperatively located. MAIN OUTCOME MEASUREMENTS: Operative findings were compared with results of preoperative studies to determine the accuracy of sestamibi SPECT and CUS for successful MIP. RESULTS: All 71 patients underwent preoperative sestamibi SPECT and CUS. Sestamibi scanning was accurate in 53 (75%) of 71 patients, whereas CUS was accurate in 40 (56%) in determining the side where the glands were located. Sestamibi scan and CUS findings were negative in 5 patients. These patients underwent planned bilateral neck exploration. Of the remaining 66 patients, MIP was successfully performed in 60 (91%). The CUS was complementary to sestamibi scanning in 9 (15%) of these 60 patients, allowing them to avoid bilateral neck exploration. CONCLUSIONS: A positive sestamibi scan finding is the only preoperative requirement for most patients with primary hyperparathyroidism for MIP. If the sestamibi scan findings are negative or ambiguous, preoperative CUS can localize an additional 14% of enlarged parathyroid glands, further facilitating an MIP in these patients. PMID- 15136351 TI - Achalasia treatment: improved outcome of laparoscopic myotomy with operative manometry. AB - HYPOTHESIS: Operative manometry detects residual esophagogastric junction (EGJ) high pressure, ensuring complete myotomy. DESIGN: Consecutive patients undergoing laparoscopic myotomy. SETTING: Tertiary care academic medical center. PATIENTS: From 1997 to 2003, 139 patients with achalasia underwent laparoscopic myotomy. INTERVENTIONS: We assessed myotomy completeness by operative endoscopy and performed operative manometry to measure pressure across the EGJ myotomy. Residual high pressure was isolated and intact muscle divided. MAIN OUTCOME MEASURES: Esophageal manometry, quality of life, and dysphagia severity score. RESULTS: Median lower esophageal sphincter pressure was 27 mm Hg preoperatively; 10 patients had sigmoid esophagus and 57 had previous dilation and/or toxin injection. There were 136 laparoscopic myotomies and 3 conversions to open procedures (2%). Operative endoscopy was performed in all patients. Operative manometry, completed in 132 patients (95%), identified residual EGJ high pressure leading to myotomy revision in 45 patients (31 in the first 70 treated). Small perforations occurred in 19 patients, associated with previous dilation and/or toxin injection in 12 patients. One-month follow-up was available in 136 patients (98%); 126 patients had minimal symptoms (93%), whereas 1 had recurrent EGJ high pressure, 5 esophagitis, 3 sigmoid esophagus, and 1 paraesophageal hernia. In 60 patients with complete 1-year follow-up, quality of life and dysphagia improved (P <.05); mean lower esophageal sphincter pressure decreased to 7.6 mm Hg (P <.05). CONCLUSIONS: Operative manometry identifies residual EGJ high pressure and reduces the incidence of incomplete myotomy. Laparoscopic myotomy improves quality of life and dysphagia symptoms and may be the treatment of choice in most patients with achalasia. PMID- 15136352 TI - Major hepatic resection for hilar cholangiocarcinoma: analysis of 46 patients. AB - HYPOTHESIS: Major hepatectomy, bile duct resection, and regional lymphadenectomy for hilar cholangiocarcinoma are associated with actual long-term (>5 years) survival. DESIGN: Retrospective outcome study. SETTING: Single tertiary referral institution. PATIENTS: Between 1979 and 1997, 46 consecutive patients had resection of hilar cholangiocarcinoma by major hepatectomy, bile duct resection, and regional lymphadenectomy. MAIN OUTCOME MEASURES: Overall survival and tumor recurrence were correlated to clinicopathological factors, operative morbidity, and mortality. RESULTS: Twenty-five patients underwent left hepatectomy, 17 underwent right hepatectomy, and 4 had extended right hepatectomy. Eighteen patients underwent resection of segment 1. Negative (R0) resection margins were achieved in 37 patients (80%). The operative mortality rate was 9%, and the surgical morbidity rate was 52%. Actual 1-year, 3-year, and 5-year survival rates were 80%, 39%, and 26%, respectively. Factors adversely associated with patient survival rates included: male sex, lymph node metastases, tumor grade 3 or 4, elevated direct serum bilirubin level at diagnosis, elevated preoperative activated partial thromboplastin time, and more than 4 U of red blood cells transfused perioperatively. Tumor size and R0 resection approached significance for survival. Factors associated with tumor recurrence included: male sex, tumor grade 3 or 4, a low hemoglobin level both at diagnosis and preoperatively, and a low preoperative prothrombin time and low alkaline phosphatase level at diagnosis and preoperatively. Median time to recurrence was 3.6 years. Tumor recurrence was predominantly local and regional. CONCLUSIONS: The actual 5-year survival rate of 26% justifies major partial hepatectomy, bile duct resection, and regional lymphadenectomy for hilar cholangiocarcinoma. The high frequency of local and regional recurrence warrants investigation of adjuvant therapy. PMID- 15136353 TI - Outcome of laparoscopic adrenalectomy for pheochromocytomas vs aldosteronomas. AB - HYPOTHESIS: Laparoscopic adrenalectomy (LA) is most commonly performed for pheochromocytomas (PHEs) and aldosteronomas (ALDs). We hypothesize that LA for these differing tumor types is associated with different operative courses and outcomes. DESIGN: Retrospective study of a 10-year experience with LA. SETTING: University teaching hospital. PATIENTS: Laparoscopic adrenalectomy was performed on 149 patients. During data analysis, the initial 35 LAs performed for various adrenal lesions were excluded to account for the learning curve. Twenty-six of 30 PHEs and 34 of 45 ALDs were included. MAIN OUTCOME MEASURES: Analysis of variance was used to compare operative time, tumor size, estimated blood loss, and postoperative length of hospital stay between the PHE and ALD groups and subsets of these groups. chi(2) Analysis was used to compare tumor location, transfusion requirements, conversion to open procedures, and incidence of major complications. RESULTS: Right-sided lesions occurred in 19 of 26 PHEs, and left sided lesions occurred in 28 of 34 ALDs (P <.001). Mean +/- SD tumor size of PHEs (4.9 +/- 1.8 cm) was larger than that of ALDs (2.7 +/- 1.7 cm) (P <.001). Mean +/ SD operative time for PHEs vs ALDs was 191 +/- 49 vs 162 +/- 48 minutes (P =.02). Mean +/- SD estimated blood loss was greater for PHEs (276 +/- 298 mL) than for ALDs (196 +/- 324 mL) (P =.33). Subset analysis revealed that the mean +/- SD size of right-sided PHEs (5.3 +/- 1.8 cm) was significantly larger than that of right-sided ALDs (3.0 +/- 1.5 cm) (P=.001). Mean +/- SD operative time for right-sided PHEs (198 +/- 44 minutes) was longer than that for right-sided ALDs (145 +/- 37 minutes) (P=.005). Six PHE patients required blood transfusions vs 2 ALD patients (P =.05). Two LAs, 1 PHE and ALD, were converted to open procedures. Mean +/- SD length of hospital stay was longer for PHE patients vs ALD patients (4 +/- 4 vs 2 +/- 3 days; P =.08). Six PHE patients had complications vs 3 ALD patients (P =.13). CONCLUSIONS: For PHEs, LA was associated with the removal of more right-sided lesions, larger tumors, longer operative times, and more complications. Trends toward greater estimated blood losses and longer hospital stays were observed for PHEs vs ALDs. Despite the advanced skills of an experienced surgeon, LA for PHEs is associated with a more complex course than for ALDs. Surgeons should begin performing LA for ALD early in their experience to avoid the potential pitfalls associated with PHEs. PMID- 15136354 TI - Neoadjuvant chemoradiotherapy for locally advanced esophageal cancer: experience at a single institution. AB - HYPOTHESIS: Patients receiving neoadjuvant chemoradiotherapy followed by surgery (CRS) undergo downstaging of their tumor and have improved survival when compared with patients undergoing surgery followed by adjuvant chemoradiotherapy (SCR). DESIGN: Retrospective study. SETTING: Tertiary-care university medical center. PATIENTS: One hundred twenty-three patients with squamous cell carcinoma and adenocarcinoma of the esophagus underwent Ivor-Lewis esophagectomy from January 1, 1990, through December 31, 2001. Of these, 31 received CRS; 27, SCR; and 65, surgery alone. INTERVENTIONS: Patients were candidates for neoadjuvant or adjuvant therapy if they had locally advanced disease (T3/T4 N0 or any T stage with N1). Neoadjuvant and adjuvant therapies were nonrandomized and based on the preference of the treating oncologist and surgeon. MAIN OUTCOME MEASUREMENTS: Pathological downstaging was analyzed in the patients receiving CRS. Operative mortality, postoperative morbidity, median survival, and overall survival were compared between the CRS and SCR groups. RESULTS: Pathological downstaging (as characterized by TNM staging) was observed in 20 (64%) of the patients receiving CRS. Complete pathological responses occurred in 5 (16%) of the patients undergoing CRS. No 30-day mortality was observed in either treatment group. No statistical difference in survival was observed between groups, although a trend suggested improved survival with neoadjuvant therapy (3-year survival in CRS and SCR groups was 45% and 22%, respectively; P =.15). Complete pathological responders in the CRS group had a 1-year survival of 80% compared with 29% in nonresponders (P =.25). No statistical differences were observed between groups in relation to blood loss, length of hospital stay, mortality, or morbidity. CONCLUSIONS: Neoadjuvant chemoradiotherapy effectively downstages cancer in patients with locally advanced esophageal disease. Morbidity and operative mortality were not significantly different between patients receiving neoadjuvant and adjuvant therapy. The difference in overall survival between the 2 groups did not reach statistical significance, although a trend at 3 years was observed. PMID- 15136355 TI - Anticoagulation is the gold standard therapy for blunt carotid injuries to reduce stroke rate. AB - HYPOTHESIS: Aggressive screening, early angiographic diagnosis, and prompt anticoagulation for blunt carotid artery injuries (CAIs) improves neurologic outcome. DESIGN: From January 1, 1996, through December 31, 2002, there were 13 280 blunt trauma admissions to our level I center, of which 643 underwent screening angiography for blunt CAI on the basis of a protocol including injury patterns and symptoms. Patients without contraindications underwent anticoagulation immediately for documented lesions. SETTING: A state-designated, level I urban trauma center. PATIENTS: Of the 643 patients undergoing screening angiography, 114 (18%) had confirmed CAI. INTERVENTION: Early angiographic diagnosis and prompt anticoagulation. MAIN OUTCOME MEASURES: Diagnosis, stroke rate, and complications stratified by method of intervention. RESULTS: A CAI was identified in 114 patients during the 7-year study period; the majority were men (71%), with a mean +/- SD age of 34 +/- 1.3 years and a mean +/- SD Injury Severity Score of 29 +/- 1.5. Seventy-three patients underwent anticoagulation after diagnosis (heparin in 54, low-molecular-weight heparin in 2, antiplatelet agents in 17); none had a stroke. Of the 41 patients who did not receive anticoagulation (because of a contraindication in 27, symptoms before diagnosis in 9, and carotid coil or stent in 5), 19 patients (46%) developed neurologic ischemia. Ischemic neurologic events occurred in 100% of patients who presented with symptoms before angiographic diagnosis and those receiving a carotid coil or stent without anticoagulation. CONCLUSIONS: Our prospective evaluation of blunt CAIs suggests that early diagnosis and prompt anticoagulation reduce ischemic neurologic events and their disability. The optimal anticoagulation regimen, however, remains to be established. PMID- 15136356 TI - Persistent acid and bile reflux in asymptomatic patients with Barrett esophagus receiving proton pump inhibitor therapy. AB - HYPOTHESIS: Symptom control does not reflect elimination of abnormal acid reflux or abnormal bile reflux in patients with long-segment Barrett esophagus receiving proton pump inhibitors (PPIs). DESIGN: Prospective survey. SETTING: University hospital. PATIENTS: Thirty-two patients with long-segment Barrett esophagus who were asymptomatic with PPIs. MAIN OUTCOME MEASURES: Twenty-four-hour ambulatory pH and bile reflux monitoring while continuing PPIs. RESULTS: Abnormal acid reflux (pH <4 for 11.9% [interquartile range, 6.8%-19.6%) of 24 hours] persisted in 15 patients (47%) who could not be distinguished from those with normal acid reflux (pH <4 for <4.5% of 24 hours) by any endoscopic, manometric, or therapeutic characteristic. Abnormal bile reflux (absorbance >0.14 for 8.7% [interquartile range, 3.9%-8.7%] of 24 hours) was detected in 11 (48%) of 23 patients, such that both normal bile reflux (absorbance >0.14 for <1.8% of 24 hours) and normal acid reflux were observed in only 8 patients (35%). There was no association between abnormal acid reflux and abnormal bile reflux. CONCLUSIONS: Despite symptom control with PPIs, both acid reflux and bile reflux were controlled in only one third of patients. Posttherapeutic monitoring of acid and bile reflux is recommended in future clinical trials of PPI treatment vs laparoscopic antireflux surgery. PMID- 15136358 TI - Ultrasound-guided minimally invasive surgery for fibroadenomas. PMID- 15136357 TI - A systematic review of the comparative safety of colloids. AB - HYPOTHESIS: Safety differences exist among colloids widely used for fluid management in acutely ill patients, as judged according to the comparative incidence of adverse events. DATA SOURCES: Colloid safety data for human subjects were sought, without language or time period restrictions, by means of computer searches of bibliographic and clinical trial databases, hand searches of medical journals and Index Medicus, inquiries with investigators and colloid suppliers, and examination of reference lists. Search terms included "colloids", "morbidity", and "mortality". STUDY SELECTION: Controlled trials, cohort studies, pharmacovigilance studies, and prior meta-analyses were independently selected by 2 unblinded investigators. Of 189 candidate studies, 113 were included, with safety data encompassing 1.54 x 10(6) patients and 1.09 x 10(8) colloid infusions. DATA EXTRACTION: Two unblinded investigators independently extracted data. Study limitations and confounding factors were tabulated. DATA SYNTHESIS: With albumin as the reference colloid, the incidence rate ratio for anaphylactoid reactions was 4.51 (95% confidence interval, 2.06-9.89) after hydroxyethyl starch administration, 2.32 (95% confidence interval, 1.21-4.45) after dextran, and 12.4 (95% confidence interval, 6.40-24.0) after gelatin. Pruritus occurrence was significantly increased by hydroxyethyl starch exposure (odds ratio, 1.78; 95% confidence interval, 1.23-2.58). Artificial colloid administration was consistently associated with coagulopathy and clinical bleeding, most frequently in cardiac surgery patients receiving hydroxyethyl starch. On the basis of large scale pharmacovigilance study results, albumin infusion resulted in a low rate of both total adverse events (3.1 to 8.6 per 10(5) infusions) and serious adverse events (1.29 per 10(6) infusions). CONCLUSIONS: Significant safety differences exist among colloids. Therefore, conclusions regarding the clinical usefulness of colloids as a fluid class should be formed with caution. PMID- 15136359 TI - Image of the month. Pseudomembranous enterocolitis. PMID- 15136360 TI - Pseudohermaphroditism. PMID- 15136361 TI - Do we really want to know why only some smokers get COPD? PMID- 15136362 TI - From "pro and con" debate to evidence-based practice: ventilator-associated pneumonia. PMID- 15136363 TI - Intensive care of patients with HIV infection: HAART warming improvement but beware of future HAART (and heart) attacks. PMID- 15136364 TI - Malignant pleural mesothelioma: the puzzling role of gene-environment interaction. PMID- 15136365 TI - Alpha1-antitrypsin deficiency: more than a protease imbalance? PMID- 15136366 TI - Rofecoxib, a COX-2 inhibitor, lowers C-reactive protein and interleukin-6 levels in patients with acute coronary syndromes. AB - BACKGROUND: Patients with acute coronary syndromes (ACS) have high levels of inflammatory mediators such as C-reactive protein (CRP) and interleukin (IL)-6. AIM: To evaluate whether patients with ACS treated with rofecoxib, a COX-2 inhibitor, will have reduced CRP, IL-6, and soluble tumor necrotic factor receptor-1 (sTNF-R1) levels and improved endothelial function. METHODS AND RESULTS: Thirty-four patients hospitalized with ACS were randomized to receive rofecoxib, 25 mg/d plus aspirin 100 mg/d, or placebo plus aspirin, 100 mg/d, for a period of 3 months. Blood samples for CRP, IL-6, and sTNF-R1 levels were drawn prior to randomization, and after 1 month and 3 months. CRP levels in the rofecoxib group (n = 18) were significantly lower both at 1 month and 3 months compared to the baseline levels (p < 0.02). IL-6 levels were significantly lower at 1 month (p < 0.02) in the rofecoxib group, but not at 3 months. There was no change in endothelial function or sTNF-R1 levels. CONCLUSION: Patients recovering from ACS had lower levels of CRP and IL-6 at 1 month and lower CRP levels at 3 months when treated with rofecoxib plus aspirin. Suppression of inflammatory processes may lead to retardation of coronary atherosclerosis and coronary events. PMID- 15136367 TI - Dosing practices and risk factors for bleeding in patients receiving enoxaparin for the treatment of an acute coronary syndrome. AB - OBJECTIVES: To describe dosing practices and to identify risk factors for bleeding in patients with an acute coronary syndrome (ACS) who received treatment with enoxaparin. DESIGN: Retrospective chart review. SETTING: Coronary care unit of a tertiary-care teaching hospital. PATIENTS: Patients with a discharge diagnosis of an ACS who received at least one dose of enoxaparin, 1 mg/kg, were eligible for this study. Enoxaparin dosing practices, factors that might influence the safety of enoxaparin administration, and bleeding events were documented. Multivariable regression analysis was used to identify independent predictors of bleeding in this clinical setting. RESULTS: Of 208 patients with an ACS who received enoxaparin, 48 patients (23%) received a dose that was > 10% or < 10% of the recommended 1 mg/kg dose, 18 patients (9%) did not have body weight documentation to guide enoxaparin dosing, and 17 patients (8%) had significant renal impairment (serum creatinine > 150 micromol/L), with the potential for bioaccumulation of enoxaparin. There were 35 bleeding events (17%), of which 8 events (4%) were major. Risk factors for any bleeding (major or minor) were increasing patient age (odds ratio [OR], 1.57; 95% confidence interval [CI], 1.13 to 2.20), coadministered nonsteroidal anti-inflammatory or antiplatelet drug therapy (OR, 2.38; 95% CI, 1.06 to 5.38), and number of enoxaparin doses (OR, 2.15; 95% CI, 1.25 to 3.68). Risk factors for major bleeding were increasing patient age (OR, 2.56; 95% CI, 1.05 to 6.28) and coadministered clopidogrel (OR, 7.70; 95% CI, 1.16 to 51.9). CONCLUSION: In this clinical practice assessment of patients with an ACS, the use of enoxaparin was suboptimal, with the potential to increase bleeding complications. Coadministered clopidogrel, other drugs that affect hemostasis, and increasing age conferred an increased bleeding risk. PMID- 15136368 TI - The potential impact of primary percutaneous coronary intervention on ventricular septal rupture complicating acute myocardial infarction. AB - BACKGROUND: Recent data suggest that the risk of acquired ventricular septal defect (VSD), a complication of acute myocardial infarction (AMI), could be reduced using thrombolytic therapy. There are, however, still no available data regarding the potential impact of primary percutaneous coronary intervention (PCI) on AMI-related VSD in a clinical setting. The purposes of this study were to delineate the incidence and the potential risk factors of AMI-related VSD in the Chinese population, and to determine whether primary PCI could reduce such risk. METHODS AND RESULTS: From May 1993 through March 2003, a total of 1,321 patients with AMI (for < 12 h) underwent primary PCI in our hospital. Of these 1,321 patients, 3 patients (0.23%) developed VSD after undergoing a primary PCI, with a mean (+/- SD) time of occurrence of 25.3 +/- 12.2 h. During the same period, a total of 616 consecutive, unselected patients with early AMI [ie, > 12 h and < or = 7 days] or recent myocardial infarction (MI) [ie, > or = 8 days and < 30 days] who had not received thrombolytic therapy underwent elective PCI. Of these 616 patients, 18 (2.9%) had VSD either on presentation or during hospitalization, with a mean time of occurrence of 71.1 +/- 64.2 h. Clinical variables were utilized to statistically analyze the potential risk factors. Univariate analysis demonstrated that the enrollment variables strongly related to this complication were advanced age, hypertension, nonsmokers, anterior infarction, female gender, and lower body mass index (BMI) [all p < 0.005]. Using multiple stepwise logistic regression analysis, the only variables independently related to VSD were advanced age, female gender, anterior infarction, and low BMI (all p < 0.05). The in-hospital mortality rate was significantly higher in patients with this complication than in patients without this complication (47.6% vs 8.0%; p < 0.0001). The incidence of this complication was significantly lower in patients with AMI who underwent primary PCI than in those with early or recent MI who underwent elective PCI (3.0% vs 0.23%, respectively; p = 0.0001). CONCLUSION: Primary PCI had a striking impact on reducing the incidence of VSD after AMI compared to elective PCI in patients who did not receive thrombolytic therapy. Advanced age, female gender, anterior infarction, and low BMI had potentially increased the risk of this catastrophic complication after AMI in this Chinese population. PMID- 15136369 TI - Mechanism of reducing platelet activity by percutaneous transluminal mitral valvuloplasty in patients with rheumatic mitral stenosis. AB - BACKGROUND: Previous studies have demonstrated that platelet activity significantly decreased after optimal percutaneous transluminal mitral valvuloplasty (PTMV) in patients with rheumatic mitral stenosis (MS). However, the mechanism of reducing platelet activity by valvuloplasty remains unclear. METHODS AND RESULTS: We studied 19 patients with symptomatic MS who were undergoing PTMV. The fractions of unstimulated platelets expressing P-selectin in the venous blood obtained before, and at the 1-week and 4-week follow-ups after PTMV were determined by flow cytometry. The mitral valve areas, measured before and at the 1-week follow-up after PTMV, were calculated by means of the Doppler pressure half-time method. The mean (+/- SD) area of the mitral valve increased significantly after PTMV (1.05 +/- 0.17 vs 1.44 +/- 0.27 cm2, respectively; p < 0.0001). The mean left atrial area was reduced in size significantly after PTMV (36.6 +/- 11.4 vs 33.9 +/- 13.4 cm2, respectively; p < 0.05). The mean left atrial pressure (23.3 +/- 5.1 vs 18.0 +/- 5.8 mm Hg, respectively; p < 0.0001) and the mean pulmonary arterial pressure (31.4 +/- 7.8 vs 26.1 +/- 7.7 mm Hg, respectively; p < 0.0001) fell significantly after PTMV. The fraction of platelets expressing P-selectin in the venous blood fell significantly after PTMV (before PTMV, 4.7 +/- 2.4%; 1 week after PTMV, 2.2 +/- 2.1%; 4 weeks after PTMV, 2.0 +/- 1.7%; p < 0.0001). Correlation analysis demonstrated that there was a significantly direct relationship between the magnitude of increase in mitral valve area and the magnitude of decrease in the fraction of platelets expressing P-selectin in the venous blood 4 weeks after PTMV (p = 0.0013; r = 0.682). However, there was no significant correlation between the magnitude of decrease in the fraction of platelets expressing P-selectin in the venous blood and the magnitude of decrease in the left atrial area, the decrease in left atrial pressure, or the decrease in the pulmonary artery pressure after PTMV. CONCLUSIONS: In patients with moderate-to-severe MS, increased platelet activation fell significantly after PTMV. It was the increase in mitral valve area by PTMV, instead of hemodynamic and echocardiographic factors, that accounted for the decrease in the fraction of venous platelets expressing P selectin after PTMV. PMID- 15136370 TI - The effect of a computerized reminder system on the prevention of postoperative venous thromboembolism. AB - STUDY OBJECTIVE: To measure the effect of an altered process of care, directed by a computerized reminder system, on rates of symptomatic postoperative venous thromboembolism. DESIGN: Comparisons of preintervention and postintervention measurements. SETTING: A university-affiliated community hospital in Utah. PATIENTS: Two-thousand seventy-seven consecutive patients who underwent major operations in four surgical divisions between January 1, 1997, and October 31, 1997 (preintervention), and 2,093 consecutive patients who underwent the same procedures between January 1, 1998, and October 31,1998 (postintervention). INTERVENTION: A program to prevent venous thromboembolism developed from American College of Chest Physicians guidelines, and an altered work process directed by a computerized reminder system. MEASUREMENTS: Rates of symptomatic, objectively confirmed deep vein thrombosis (DVT), pulmonary embolism (PE), and death attributable to venous thromboembolism occurring within 90 days of the date of surgery. RESULTS: The preintervention and postintervention cohorts did not differ with respect to age, severity of illness, number of risk factors for venous thromboembolism, or individual risk factors for venous thromboembolism. The overall prophylaxis rate increased from 89.9% before implementation of the computerized reminder system to 95.0% after implementation (p < 0.0001). The combined 90-day rate of symptomatic DVT, PE, and death attributable to PE remained the same (preintervention, 1.0%; postintervention, 1.2%; odds ratio, 1.21; 95% confidence interval, 0.67 to 2.20). Forty of 46 venous thromboembolic complications (87%) occurred despite the delivery of American College of Chest Physicians-recommended measures to prevent venous thromboembolism. CONCLUSIONS: Computerized reminder systems combined with altered care procedures increase the rate of prophylaxis against venous thromboembolism without decreasing the rate of symptomatic venous thromboembolism when the baseline rate of prophylaxis is high. A population of surgical patients exists who are resistant to American College of Chest Physicians-recommended prophylactic measures against venous thromboembolism. New strategies are needed to address prophylaxis-resistant venous thromboembolism. PMID- 15136371 TI - Costs and clinical outcomes associated with low-molecular-weight heparin vs unfractionated heparin for perioperative bridging in patients receiving long-term oral anticoagulant therapy. AB - STUDY OBJECTIVES: There have been no health-care cost evaluations comparing the use of low-molecular-weight heparin (LMWH) to unfractionated heparin (UH) as "bridge therapy" in the perioperative period in patients receiving long-term oral anticoagulant (OAC) therapy who need interruption of therapy to undergo an elective surgical procedure. We performed a retrospective analysis of the medical and administrative records of health plan members in a managed care organization who underwent bridge therapy perioperatively with either i.v. UH, administered in a hospital setting, or LMWH, administered primarily in the outpatient setting using disease management guidelines. DESIGN: A retrospective analysis of medical and administrative records of treated health plan members meeting inclusion/exclusion criteria during the two study periods (ie, from 1994 to 1996 and from 1998 to 2000). SETTING: Staff-model health maintenance organization serving New Mexico. PATIENTS: The UH group included persons receiving long-term warfarin therapy from 1994 to 1996 (26 patients), and the LMWH group included persons receiving long-term warfarin therapy from 1998 to 2000 (40 patients) with perioperative use of heparin (either UH or LMWH) as bridge therapy for an elective surgical procedure. INTERVENTIONS: Costs were calculated for the period from 10 days before the procedure through 30 days after the procedure. The rates of adverse events (ie, valvular or mural thrombus, intracranial event, transient ischemic attack, peripheral arterial event, venous thromboembolic event, major and minor bleeding, thrombocytopenia, and death) occurring 1 to 30 days postprocedure were determined. MEASUREMENTS AND RESULTS: The groups were similar in age, sex, Charlson score, indication for long-term warfarin therapy (ie, arterial/cardiac vs venous), mean international normalized ratio prior to procedure, procedure duration, use of intraprocedural anticoagulant agents or thrombolytic agents, and use of general anesthesia during the procedure (all p > 0.05). A total of 34.6% of UH patients and 40.0% of LMWH patients experienced one or more clinical adverse events within 30 days of the postoperative period, a difference that was not statistically significant (p = 0.67). The mean total health-care costs were 31,625 dollars in the UH group and 18,511 dollars in the LMWH group (p < 0.01). The mean inpatient costs were 28,515 dollars in the UH group and 14,330 dollars in the LMWH group (p < 0.01). Outpatient surgery costs (1,159 dollars vs 53 dollars, respectively; p = 0.01) and pharmacy costs (639 dollars vs 133 dollars, respectively; p < 0.01) were higher in the LMWH group. CONCLUSIONS: The mean total health-care costs in the perioperative period were significantly lower (by 13,114 dollars) in patients receiving long-term OAC therapy using LMWH compared to those receiving it using UH for an elective surgical procedure. The cost savings associated with LMWH use were accomplished through the avoidance or minimization of inpatient stays and no increase in the overall rate of clinical adverse events in the postoperative period. PMID- 15136372 TI - ECG score predicts those with the greatest percentage of perfusion defects due to acute pulmonary thromboembolic disease. AB - BACKGROUND: More aggressive management may be warranted for patients with acute pulmonary embolism (PE) and the greatest pulmonary vascular obstruction. We hypothesized that a scoring system based on the ECG might identify such patients. METHODS: Consecutive patients investigated for PE at Christchurch Hospital between 1997 and 2002 with high-probability ventilation/perfusion (V/Q) scan findings were studied. The ECG obtained closest to and within 48 h of the scan was scored by two independent observers, and the mean ECG score was calculated. V/Q scan findings were categorized into those with < 30%, 30 to 50%, and > 50% perfusion defect by two independent observers experienced in V/Q interpretation. A consensus score was taken when disagreement occurred. RESULTS: Two hundred twenty-nine patients were included in the study. The interobserver agreement for ECG score was 0.96 (Cronbach alpha) and V/Q score was 0.55 (kappa). The ECG predicted those with the greatest amount of perfusion defects. Mean ECG score was 2.6 (SD 2.8) in patients with < 30% perfusion defect, 3.2 (SD 2.9) in patients with 30 to 50% perfusion defect, and 5.3 (SD 3.7) in patients with > 50% perfusion defect. The area under the receiver operating characteristic curve for ECG score and those with > 50% perfusion defect was 0.71 (SE 0.04). An ECG score of > or = 3 predicted those with > 50% perfusion defect with a sensitivity of 70% (95% confidence interval [CI], 59 to 81%), and a specificity of 59% (95% CI, 51 to 67%). CONCLUSION: An ECG score, simple to derive, predicts those with the greatest percentage of perfusion defect. Using the ECG for management warrants prospective evaluation. PMID- 15136373 TI - Geographic and gender variability in the prevalence of bronchial responsiveness in Canada. AB - OBJECTIVES: Geographic variability in reported prevalences of asthma worldwide could in part relate to interpretation of symptoms and diagnostic biases. Bronchial responsiveness measurements provide objective evidence of a common physiologic characteristic of asthma. We measured bronchial responsiveness using the standardized protocol of the European Community Respiratory Health Survey (ECRHS) in six sites in Canada, and compared prevalences across Canada with international sites. DESIGN: Samples of 3,000 to 4,000 adults aged 20 to 44 years were randomly selected in Vancouver, Winnipeg, Hamilton, Montreal, Halifax, and Prince Edward Island, and a mail questionnaire was completed by 18,616 individuals (86.5%). Preselected random subsamples (n = 2,962) attended a research laboratory for examination including more detailed questionnaires, lung function testing including methacholine challenge, and skin testing with 14 allergens. RESULTS: Prevalences of bronchial hyperresponsiveness, measured as cumulative dose of methacholine required to produce a 20% fall from the post saline solution FEV1 < or = 1 mg, ranged from 4.9% (95% confidence interval [CI], 1.6 to 8.5) in Halifax to 22.0% (95% CI, 18.1 to 26.0) in Hamilton (median, 10.7%). In all Canadian sites, bronchial hyperresponsiveness was more prevalent in women than in men. Neither the geographic nor gender differences were accounted for by differences in age, smoking, skin test reactivity, or baseline FEV1. Geographic- and gender-related variability changed little when only bronchial hyperresponsiveness associated with asthma-like symptoms was considered. CONCLUSIONS: A wide variability in bronchial responsiveness can occur within one country, almost as wide as the range found across all international sites participating in the ECRHS study and not explained by differences in gender, smoking, skin test reactivity, and FEV1. While gender variability in the prevalence of bronchial responsiveness is likely due to hormonal and immunologic factors, geographic variability is likely to result from environmental factors. PMID- 15136374 TI - Prevalence of asthma and chronic respiratory symptoms among Alaska Native children. AB - STUDY OBJECTIVES: To quantify the prevalence and impact of chronic respiratory symptoms among predominantly Alaska Native (AN)/American Indian (AI) middle school students. DESIGN: School-based prevalence assessment using the International Study of Asthma and Allergy in Children survey, with supplemental video material and added questions about productive cough, exposure to tobacco smoke, and the functional impact of symptoms. SETTING: The Yukon-Kuskokwim delta region of western Alaska. PARTICIPANTS: A total of 466 children in the sixth to ninth grades, 81% of whom are AN/AI (377 children). INTERVENTIONS: No study intervention. RESULTS: Among the 377 AN/AI children, 40% reported one of the following three categories of chronic respiratory disease: physician-diagnosed asthma, 7.4%; asthma-like symptoms (ALS) without an asthma diagnosis, 11.4%; and chronic productive cough (CPC) without asthma diagnosis or symptoms, 21.5%. Symptom prevalence differed substantially between the largest town in the region and rural villages. After an adjustment for demographic factors, exposure to environmental tobacco smoke, active tobacco smoking, and self-report of atopy, village residents were 63% less likely to have ALS (p = 0.009), and had a twofold greater risk of CPC (p < 0.001) compared to children living in the town. Children with respiratory symptoms experienced sleep disturbances and accessed clinic visits for respiratory problems more often than did asymptomatic children. CONCLUSIONS: Chronic respiratory symptoms are very common among AN children. CPC is an important nonasthmatic respiratory condition in this population. The differing patterns of respiratory illness within this region may help to elucidate the specific risk factors for asthma and chronic bronchitis in children. PMID- 15136375 TI - Effects of a comprehensive school-based asthma program on symptoms, parent management, grades, and absenteeism. AB - STUDY OBJECTIVE: This study assessed the impact of a comprehensive school-based asthma program on symptoms, grades, and school absences in children, and parents' asthma management practices. DESIGN: Randomized controlled trial. SETTING: Fourteen elementary schools in low-income neighborhoods in Detroit, MI. PARTICIPANTS: Eight hundred thirty-five children with asthma in grades 2 through 5 and their parents. INTERVENTION: The intervention entailed six components for children, their parents, classmates, and school personnel to encourage and enable disease management. MEASUREMENTS AND RESULTS: Parents completed telephone interviews and the schools provided data at baseline and 24 months after intervention. At follow-up, treatment children with persistent disease had significant declines in both daytime (14% fewer, p < 0.0001) and nighttime (14% fewer, p < 0.0001) symptoms. Among children with both mild intermittent and persistent disease, those in the treatment group had 17% fewer daytime symptoms (p < 0.0001) but 40% more nighttime symptoms. Treatment children had higher grades for science (p < 0.02) but not reading, mathematics, or physical education. No differences in school absences for all causes between groups were noted in school records. However, parents of treatment group children reported fewer absences attributable to asthma in the previous 3 months (34% fewer, p < 0.0001) and 12 months (8% fewer, p < 0.05). Parents of treatment children had higher scores (2.19 greater, p = 0.02) on an asthma management index. The program may have stimulated attention to symptoms at night by parents of children with mild intermittent disease. Overall, the intervention provided significant benefits, particularly for children with persistent asthma. PMID- 15136376 TI - Variations in pediatric asthma hospitalization rates and costs between and within Nordic countries. AB - BACKGROUND: We assessed variations in hospitalization parameters and costs among asthmatic children in four Nordic countries by geographic location and age groups. METHODS: Cross-sectional, county-level aggregate data on asthma-related hospitalizations in 1999, obtained from public national databases for children < 15 years old from Denmark, Sweden, Norway, and Finland, together with country specific asthma management cost were used to estimate the incidence of first hospital admission (per 1,000), length of hospital stay (LOS), and hospitalization cost. Longitudinal patient-specific data from 1998/1999 were used to calculate the relative hazard of readmission (RHR) using a multivariate Cox proportional hazards model. RESULTS: Nordic incidence of first hospital admission in 1999 was 2.17 per 1,000 children, readmission was noted in 16% of the patients, mean LOS was 2.64 days, and total hospitalization cost was almost 14 million dollars. Hospitalization incidence, RHR, and costs were significantly higher in children < 5 years old compared with school children 6 to 14 years old. Hospital LOS, incidence of first hospital admission, and cost per child were the highest in Denmark, though RHR did not differ significantly from Sweden. CONCLUSIONS: Large variations in all parameters were observed between and within countries. Given the similarities among the four countries studied, these results may, among other reasons, indicate different efficiencies of the various asthma management plans between and within them. The presented measures of hospitalization patterns could prove to be valuable quality-of-care measures to guide further improvements in asthma management. PMID- 15136377 TI - History of ear infections and prevalence of asthma in a national sample of children aged 2 to 11 years: the Third National Health and Nutrition Examination Survey, 1988 to 1994. AB - METHODS: We analyzed data on 7,538 children aged 2 to 11 years who participated in the Third National Health and Nutrition Examination Survey to examine the cross-sectional associations of history of ear infections with prevalence of ever diagnosed asthma and the prevalence of wheezing in the last year in US children. RESULTS: History of ear infections was significantly related to the lifetime prevalence of diagnosed asthma (prevalence odds ratio [POR], 1.57; 95% confidence interval [CI], 1.05 to 2.36) and to the prevalence of wheezing in the last year (POR, 1.70; 95% CI, 1.22 to 2.37) after controlling for potential confounding variables. The number of ear infections was linearly and significantly related to the risk of asthma and wheezing in the last year. Among children with no diagnosis of asthma, there was a significant association between a history of ear infections and any wheezing in the last year (adjusted POR, 1.55; 95% CI, 1.07 to 2.25). CONCLUSIONS: Our study indicated strong and significant associations of a history of asthma and wheezing with the frequency of ear infections in a nationally representative sample of 7,538 children aged 2 to 11 years. These findings highlight the need for prospective studies to examine further the relationship between asthma and ear infections. PMID- 15136378 TI - Leukotriene modifier vs inhaled corticosteroid in mild-to-moderate asthma: clinical and anti-inflammatory effects. AB - STUDY OBJECTIVE: Evidence for the anti-inflammatory activity of leukotriene receptor antagonists in humans is somewhat limited. There are also no data comparing the anti-inflammatory effects of leukotriene receptor antagonists with those of inhaled corticosteroids. This study was designed to assess the clinical efficacy and anti-inflammatory effects of leukotriene receptor antagonist plus low-dose inhaled corticosteroids compared to those of a high-dose inhaled corticosteroid in patients with mild-to-moderate asthma. METHODS: Forty-nine patients with newly diagnosed asthma were recruited. They were randomly assigned to groups that received, for a 6-week period, either (1) budesonide, 600 microg bid (1,200 microg/d) or (2) budesonide, 200 microg (400 microg/d), and zafirlukast, 20 mg bid. The variables of asthma control were recorded daily. Sputum induction and methacholine provocation tests were performed. RESULTS: The results indicated that the administration of a low-dose inhaled corticosteroid plus zafirlukast was as effective as that of a high-dose inhaled corticosteroid regarding clinical improvement and anti-inflammatory effects (ie, eosinophil percentage, and eosinophilic cationic protein [ECP] and cysteinyl leukotriene C4 levels in induced sputum). Nineteen (group 1, 8 patients; group 2, 11 patients) of 49 patients (38.8%) had returned to normal airway responsiveness after treatment. Among these patients, 16 patients (84.2%) had normal ECP levels and 10 patients (52.6%) had normal percentages of eosinophils. ECP level, but not the eosinophil percentage, was significantly associated with symptom scores. The peak expiratory flow rate (PEFR) showed a significant correlation with the provocative concentration of methacholine causing a 20% fall in FEV1 (PC20) instead of with symptom scores. CONCLUSIONS: The addition of a leukotriene modifier to treatment with low-dose inhaled corticosteroids is equivalent to treatment with high-dose inhaled corticosteroids in patients with newly diagnosed mild-to-moderate asthma. In addition to symptoms and PEFR, the monitoring of ECP and PC20 may be of great value in achieving optimal control of asthma. PMID- 15136379 TI - Effects of pranlukast administration on vascular endothelial growth factor levels in asthmatic patients. AB - STUDY OBJECTIVES: We have previously found that vascular endothelial growth factor (VEGF) levels in induced sputum were increased in asthmatic patients, and that its levels were closely associated with the degree of airway obstruction and microvascular permeability. Therefore, this study was designed to examine the effects of pranlukast, a selective leukotriene receptor antagonist, on VEGF levels in induced sputum from steroid-untreated or steroid-treated asthmatic patients. DESIGN: Double-blind, randomized, placebo-controlled, crossover study. SETTING: University hospital. PARTICIPANTS: Twenty-three asthmatic patients (steroid-untreated, 13 patients; steroid-treated, 10 patients) and 10 healthy control subjects. INTERVENTIONS: All asthmatic patients received 4-weeks of therapy with pranlukast (225 mg bid), and sputum induction was performed before and after the 4-week treatment course. MEASUREMENTS AND RESULTS: In steroid untreated asthmatic patients, the mean percentage of eosinophils (%EOS) and mean eosinophil cationic protein (ECP) levels in induced sputum were significantly decreased after 4 weeks of pranlukast administration (%EOS: before, 16.7% [SD, 7.1%]; after, 12.3% [SD, 4.0%]; p = 0.03; ECP levels: before, 774 ng/mL [SD, 258 ng/mL]; after, 564 ng/mL [SD, 204 ng/mL]; p = 0.034). Moreover, VEGF levels in the induced sputum and the airway vascular permeability index also were decreased after pranlukast administration (VEGF levels: before, 5,670 pg/mL [SD, 1,780 pg/mL]; after, 4,380 pg/mL [SD, 1,540 pg/mL]; p = 0.026; airway vascular permeability index: before, 0.032 [SD, 0.012]; after, 0.017 [SD, 0.006]; p = 0.01). In addition, the change in airway vascular permeability index from before to after pranlukast administration was significantly correlated with the change in VEGF levels (r = 0.782; p = 0.007). However, in steroid-treated asthmatic patients there was no significant difference in mean VEGF levels in induced sputum between placebo administration (before, 3,640 pg/mL [SD, 1,020 pg/mL]; after, 3,640 pg/mL [SD, 960 pg/mL] and pranlukast administration (before, 3,660 pg/mL [SD, 940 pg/mL]; after, 2,950 pg/mL [SD, 890 pg/mL]). CONCLUSIONS: Pranlukast administration decreased airway microvascular permeability through, at least in part, a decrease in airway VEGF levels in steroid-untreated asthmatic patients. However, it is likely that pranlukast administration added little efficacy to inhaled corticosteroid therapy for reduction in airway VEGF levels. PMID- 15136380 TI - Activation of bronchial epithelial cells in smokers without airway obstruction and patients with COPD. AB - STUDY OBJECTIVE: The aim of this study was to investigate the basal level as well as the tumor necrosis factor (TNF)-alpha- and interferon (IFN)-gamma-induced expression and release of the neutrophil chemoattractants interleukin (IL)-8 and growth-related oncogene (GRO)-alpha in primary bronchial epithelial cells (PBECs) from smokers without airflow obstruction and patients with COPD. In addition, the expression of both TNF-alpha-receptor subtypes--p55 TNF-receptor subtype (TNF R55) and p75 TNF-receptor subtype (TNF-R75)--was quantified in PBECs. DESIGN: PBECs from eight smokers without airflow limitation and eight patients with COPD were stimulated with 50 ng/mL of TNF and 200 U/mL of IFN-gamma for 4 h along with unstimulated time controls. The transcriptional expression and protein release were quantitatively assessed by means of real-time polymerase chain reaction and enzyme-linked immunosorbent assay. RESULTS: Basal level messenger RNA (mRNA) expression and protein release of IL-8 and GRO-alpha were not significantly different between both groups, although a trend toward higher IL-8 levels was seen in patients with COPD. TNF-alpha induced significantly higher mRNA amounts of IL-8 (p = 0.005) and GRO-alpha (p = 0.007) in patients with COPD. This was accompanied by higher protein release data for IL-8 (p = 0.005) and GRO-alpha (p = 0.007). IFN-gamma had no significant effect on the mRNA expression and protein release of IL-8 and GRO-alpha in either group. TNF-R55 and TNF-R75 were detectable in PBECs. However, no significant differences were found between both groups with respect to steady-state mRNA levels of TNF-alpha-receptor subtypes. CONCLUSION: PBECs from patients with COPD show significantly higher TNF-alpha induced release of the neutrophil chemoattractant CXC-chemokines IL-8 and GRO alpha compared to smokers without airflow limitation. This increased activation of PBECs may contribute to the predominance of neutrophils seen in the airway lumen of patients with COPD. PMID- 15136381 TI - Assessment of emphysema in COPD: a functional and radiologic study. AB - OBJECTIVES: A combination of functional measurements reflecting a decrease in maximum flow, a degree of lung hyperinflation, the relationship between maximum inspiratory and expiratory flows, bronchodilator response, and diffusing capacity of the lung for carbon monoxide (DLCO) was used to quantify the extent of emphysema, as assessed by high-resolution CT (HRCT) scanning. DESIGN: Forced inspiratory and expiratory spirometry, lung volumes, reversibility test, and single-breath diffusing capacity were assessed before and after inhaling albuterol, 200 microg. Relationships between lung function variables and emphysema extent, as determined by HRCT scanning, were tested by univariate and multivariate analyses. SUBJECTS: Thirty-nine COPD outpatients with moderate-to severe obstruction. MEASUREMENTS AND RESULTS: Emphysema extent, as assessed by HRCT scanning, ranged from 18 to 70%. All of the lung function parameters that were studied, except for the change in FEV1 percent predicted after salbutamol inhalation, correlated significantly with the extent of emphysema (r2 range, 0.19 to 0.44). Functional residual capacity, forced expiratory flow at 50% of FVC/forced inspiratory flow at 50% of FVC, DLCO/alveolar volume ratio, and bronchodilator-induced change in FEV1/FVC ratio were the only variables retained by stepwise multiple regression analysis. The multiple regression model explained 71% of the variability of emphysema extent measured by HRCT scanning. CONCLUSIONS: The combination of lung function measurements reflecting lung hyperinflation, bronchial collapsibility, lung diffusing capacity, and bronchodilator response provides a good estimate of the extent of emphysema, as evaluated by HRCT scanning. These data suggest that pulmonary function tests are useful in assessing and monitoring parenchymal damage in COPD patients. PMID- 15136382 TI - Obstructive lung disease among the urban homeless. AB - STUDY OBJECTIVES: Homelessness is a growing problem in the United States that may significantly impair physical health. The homeless have a high prevalence of cigarette smoking, poor nutrition, and adverse environmental exposures, which could contribute to obstructive lung disease (OLD). Despite this risk, the prevalence of OLD among the homeless remains unknown. We aimed to systematically assess the prevalence of OLD among the urban homeless. DESIGN, SETTING, AND PARTICIPANTS: We conducted a cross-sectional study of the prevalence of OLD among homeless individuals in San Francisco. By random sampling, we recruited 68 adults living in one homeless shelter to participate in a structured interview survey and spirometry assessment. We used a multifaceted approach to assess OLD, including respiratory symptoms, self-reported physician diagnosis of asthma, chronic bronchitis, emphysema, or COPD, and spirometry (defined as FEV1 < 80% predicted and FEV1/FVC ratio < 0.70). RESULTS: Sixty-eight adults completed the survey, and 67 adults completed the spirometry. Homeless adults were likely to be homeless < 1 year and homeless for the first time. There was a high prevalence of cigarette smoking (75% ever smokers, 68% current smokers). The prevalence of symptoms suggestive of OLD was high, including cough (29%), wheezing (40%), chronic bronchitis symptoms (21%), and dyspnea on exertion (29%). A substantial proportion of homeless subjects indicated a prior diagnosis of asthma (24%), chronic bronchitis (19%), and COPD (4%). Based on spirometry, the prevalence of OLD was 15% (95% confidence interval, 8 to 26%), which was more than double the expected prevalence in the general US population. CONCLUSIONS: As OLD is a leading cause of death in the United States, it is important to identify it early for treatment. Homeless individuals have a higher-than-expected prevalence of OLD. Public health interventions should target the homeless population for prevention and treatment of OLD. PMID- 15136383 TI - The effect of inhaled tiotropium bromide on lung mucociliary clearance in patients with COPD. AB - STUDY OBJECTIVE: To assess the effects of tiotropium on lung mucociliary clearance in COPD. DESIGN: Randomized, double-blind, placebo-controlled, parallel group study. SETTING: Outpatients of an urban-area university teaching hospital. PATIENTS: Thirty-four patients with COPD aged 40 to 75 years classified equally into two groups. INTERVENTION: Single (18 microg) daily dose of tiotropium inhalation capsules or of placebo for 21 days. METHODS: Six-hour tracheobronchial clearance of inhaled 99mTc-labeled polystyrene particles using a 48-h retention measurement to determine the "nontracheobronchial" deposition fraction. RESULTS: Test radioaerosol penetration into the lungs increased significantly (p < 0.003) as did FEV1 (p < 0.006) in the tiotropium-treated patients, but measured mucociliary clearance was not significantly changed despite the increased pathway length for clearance (mean +/- SE area under the tracheobronchial retention curve changed from 442 +/- 22 to 453 +/- 20%/h). Smaller (nonsignificant) decreases of radioaerosol penetration and FEV1 occurred in the placebo group together with a small (nonsignificant) decrease in the area under the retention curve. CONCLUSION: Twenty-one days of inhaled tiotropium, 18 microg/d, as a dry powder does not retard mucus clearance from the lungs. PMID- 15136384 TI - Actual and predicted postoperative changes in lung function after pneumonectomy: a retrospective analysis. AB - STUDY OBJECTIVES: Little is known about long-term effects of pneumonectomy on lung function and exercise tolerance. We evaluated the long-term validity of two formulas frequently used to predict postoperative lung function, as well as trends in postoperative lung function and late postoperative exercise capacity. SETTING: Nonuniversity teaching hospital of Eindhoven, the Netherlands. PATIENTS: Patients who underwent pneumonectomy between 1993 and 1998 and survived for > 1 year after the operation. MEASUREMENTS AND RESULTS: Lung function and exercise test data of 32 patients were analyzed. Postoperative FVC and FEV1 according to Kristersson/Olsen (split function of resected lung) and Juhl and Frost (number of segments to be resected) were calculated and compared with observed values measured in the third postoperative year. Calculated values correlated well with observed values, whereas Kristersson/Olsen appeared to be more accurate than Juhl and Frost. When considering trends in FEV1, we found a mean decline of 44 mL/yr; only three patients (12%) showed a rapid decline of > 100 mL/yr. Of 14 patients (44%), postoperative maximal exercise capacity was impaired due to ventilatory limitation. CONCLUSIONS: The Kristersson/Olsen formula was more accurate in predicting postoperative lung function in the third postoperative year in pneumonectomy patients. Although the annual decline in FEV1 in these patients is almost the same as in healthy patients without COPD, pneumonectomy has serious implications on exercise capacity in many patients. PMID- 15136385 TI - Is major pulmonary resection by video-assisted thoracic surgery an adequate procedure in clinical stage I lung cancer? AB - OBJECTIVE: Although several studies have shown that video-assisted thoracic surgery (VATS) for major pulmonary resection is less invasive than open thoracotomy, VATS for lung cancer has been performed in only a limited number of institutions. We aimed to review our experience of VATS for major pulmonary resections, and to determine its safety and adequacy in stage I lung cancer. METHODS: Between August 1999 and March 2003, we performed major pulmonary resection by VATS in 106 patients with lung cancer and preoperatively determined clinical stage I disease. We evaluated the number of procedures converted to open thoracotomy and the reasons for conversion, the intraoperative blood loss, interval between surgery and chest tube removal, length of postoperative hospital stay, postoperative complications, mortality rate, prognoses, and patterns of recurrence. RESULTS: We successfully performed VATS in 95 patients, whereas in another 11 patients (10%) conversion to open thoracotomy was required. The operative procedures were lobectomy in 86 patients, segmentectomy in 8 patients, and bilobectomy in 1 patient. In 95 patients who underwent VATS, postoperative complications developed in 9 patients (9%), and 1 patient (1%) died from pneumonia. In the 86 patients without complications, the mean postoperative hospital stay was 7.6 days (range, 4 to 15 days). In a mean follow-up period of 25 months (range, 6 to 48 months) in patients with non-small cell lung cancer (NSCLC), including the one perioperative death, the 3-year survival rate was 93% in 82 patients with clinical stage I disease, and 97% in 68 patients with pathologic stage I disease. The 3-year disease-free survival rate was 79% in patients with clinical stage I disease, and 89% in patients with pathologic stage I disease. Local recurrence was observed in six patients (6%): recurrence in mediastinal lymph nodes in five patients, and in the bronchial stump in one patient. CONCLUSIONS: Major pulmonary resection by VATS is acceptable in view of its low perioperative mortality and morbidity, and is an adequate procedure for the achievement of local control and good prognosis in patients with clinical stage I NSCLC. PMID- 15136386 TI - CT fluoroscopy-guided bronchoscopic dye marking for resection of small peripheral pulmonary nodules. AB - STUDY OBJECTIVE: To determine the diagnostic reliability and safety of a new marking technique using transbronchoscopic dye injection under CT fluoroscopy for preoperative localization of a small pulmonary nodule. DESIGN: Prospective study. SETTING: Hyogo Medical Center for Adults and Shizuoka Cancer Center in Japan. PATIENTS: Seventeen patients who had a peripheral pulmonary nodule < 15 mm in size on CT scans that was suspected to be difficult to localize by visual inspection and manual palpation at our institutes between April 2000 and October 2002. INTERVENTIONS: After a bronchoscope was inserted orally under local anesthesia and was introduced into the related bronchus of the target nodule, a Teflon sheath catheter with metal tip was inserted transbronchoscopically and was advanced into the visceral pleura. By monitoring CT fluoroscopy, the catheter tip was positioned at the nearest pleural surface of the nodule, and 0.5 mL indigo carmine was injected under deep inspiratory breathhold. CT scans were obtained to confirm the relationship between the injected dye area and the nodule. MEASUREMENTS AND RESULTS: The dye injections were performed completely in all 17 patients, who subsequently underwent lung resection guided by the dye staining. There were no complications or harmful effects of the surgery. The area of injected dye was demonstrated as a hazy focal lesion about 10 mm beneath the pleura on the high-resolution CT scan, and was clearly visible as a patchy dark blue area about 20 mm in size on the visceral pleura at surgery. The mean distance between the nodule and the dye was 20 mm on the CT scan (distance range, 0 to 30 mm). The mean examination time with this technique was approximately 35 min (range, 25 to 45 min). The mean CT fluoroscopic time was 60 s (range, 30 to 120 s). CONCLUSIONS: Our transbronchial "tattooing" technique is safe and reliable. We think it is superior to previous marking methods. PMID- 15136387 TI - Use of a clinical pathway to manage unsuspected radiographic findings. AB - STUDY OBJECTIVES: To describe our 5-year experience with a clinical pathway used to ensure the timely communication and evaluation of unsuspected radiologic findings (URFs) noted on clinically requested chest imaging. DESIGN: Prospective data collection on clinical practice. SETTING: Academically affiliated Veterans Affairs medical center. PARTICIPANTS: Pulmonary physicians, nurses, and radiologists. RESULTS: Over a period of 5 years, 1,629 URFs were referred to the pathway (from chest radiographs, 1,359 [83.4%]; from CT scans, 270 [16.6%]). Most URFs (78%) were nodules, with a specific diagnosis made in one third of URFs, and with a specific diagnosis thought to be clinically significant in another one third of URFs. The most common diagnosis was neoplasm, with over two thirds of these diagnoses being lung cancer. One third of lung cancers detected were either stage 1 or 2, with 1 in 17 of all URFs being stage IA lung cancer. The cost of the pathway was estimated at 28,600 dollars per year. CONCLUSIONS: URFs noted on chest imaging are frequently clinically significant, and a systematic approach to managing URFs, such as a clinical pathway, can significantly improve care in a large teaching hospital. PMID- 15136388 TI - Mandibular advancement titration for obstructive sleep apnea: optimization of the procedure by combining clinical and oximetric parameters. AB - BACKGROUND: Oral appliances (OAs) have been used for the treatment of obstructive sleep apnea syndrome (OSAS), with different degrees of effectiveness having been shown in previous studies. But, in the absence of a consensual recommendation, the method of the determination of effective mandibular advancement varies from one study to another. STUDY OBJECTIVE: We prospectively evaluated an OA titration protocol based on a combined analysis of symptomatic benefit and oximetric recording to guide the progressive mandibular advancement. SETTING: University hospital sleep disorders center. PATIENTS: Forty patients with OSAS (mean [+/-SD] apnea-hypopnea index [AHI], 46 +/- 21 events per hour) found on baseline polysomnography, who were intolerant of nasal continuous positive airway pressure, completed all aspects of the study. METHODS: Two acrylic appliances connected by Herbst attachments were constructed. The mandible was advanced 1 mm every week until there was a resolution of the symptoms and a reduction in the oxygen desaturation index (ie, the number of desaturations yielding a > 3% fall in pulse oximetric saturation per hour of recording) [ODI] of <10 events per hour of recording or a maximum comfortable limit of advancement was obtained. The final response to OA was evaluated by full polysomnography recording. RESULTS: A complete response (ie, mean AHI, 5 +/- 3 events per hour; mean snoring reduction [SR], 91 +/- 13%; mean Epworth sleepiness scale [ESS] score, 5 +/- 3) was obtained in 63.6% of patients, and a limited response (ie, mean AHI, 21 +/- 11 events per hour; mean SR, 88 +/- 15%; mean ESS, 6 +/- 3) was obtained in 18.2% of patients. Twenty-five percent of mandibular advancements were motivated by an abnormal ODI (ie, 21 +/- 10 events per hour) despite resolution of the symptoms, while 20% were motivated by persistent symptoms with a normal ODI (ie, 6 +/- 2 events per hour). After a mean duration of 17 +/- 4 months, 34 patients declared that they had used the OA 5 +/- 2 days a week for 89 +/- 19% of their sleep time. CONCLUSIONS: A combination of the patient's subjective evaluation and oximetric score improves the effectiveness of the OA titration procedure. PMID- 15136389 TI - The effects of nasal continuous positive airway pressure on platelet activation in obstructive sleep apnea syndrome. AB - OBJECTIVE: A case-controlled study to assess the effects of nasal continuous positive airway pressure (CPAP) on platelet activation in patients with obstructive sleep apnea (OSAS) syndrome. METHODS: We recruited 65 patients with suspected OSAS for this study. Blood samples were taken with the patient in the supine position in the morning immediately after polysomnography, and 1 night and 3 months after the start of nasal CPAP therapy to measure an index of platelet activation (IPA+), which reflected both the quantity and quality of platelet activation. Significant OSAS was defined as an apnea-hypopnea index (AHI) of > or = 10 events per hour. RESULTS: There were 42 patients with significant OSAS and 23 control subjects with AHI < 10 events per hour. The mean (+/- SD) age for the OSAS patients was 48 +/- 9 years, the mean body mass index was 30.7 +/- 4.8, the mean AHI was 47 +/- 25 events per hour, the mean arousal index (AI) was 37 +/- 23 events per hour, and the mean minimum arterial oxygen saturation was 74 +/- 11%. Following multiple linear regression analyses of the clinical and polysomnography parameters, AI was the independent factor that correlated best with the baseline IPA+ (beta-coefficient, 0.386; p = 0.006). Following nasal CPAP treatment with a mean objective CPAP compliance of 3.9 +/- 1.9 h per night, there was a significant decrease in IPA+ from 15.1 +/- 12.2 U (at baseline) to 12.2 +/- 5.2 U (p < 0.001) and 9.8 +/- 4.3 U (p = 0.005), respectively, after 1 night and 3 months, whereas no significant change was noted among the control subjects. Using univariate analysis of variance to compare the changes in IPA+ between the two groups at 3 months with adjustment for the baseline value, nasal CPAP reduced IPA+ by 5.63 (SE, 1.85), whereas IPA+ increased in control subjects by 1.33 (SE, 1.27) [least-squared mean difference between groups, 3.34; 95% confidence interval, 0.42 to 6.26; p = 0.026]. CONCLUSIONS: OSAS, through repeated episodes of arousals, may lead to platelet activation, which can be reduced by nasal CPAP therapy. PMID- 15136390 TI - Percutaneous multiple-site parietal pleural biopsy: description and evaluation of a new and safe technique. AB - STUDY OBJECTIVES: (1) To describe a new percutaneous pleural biopsy technique to obtain multiple-site parietal pleural biopsy specimens in patients with pleural effusion (PE), and (2) to evaluate its effectiveness and safety compared to current techniques. DESIGN: Prospective interventional study. SETTING: University teaching hospital. PATIENTS: Consecutively referred for evaluation of exudative PE. INTERVENTION: With the patient in a semirecumbent position, a 9F sheath was inserted by the Seldinger technique into the pleural cavity on the midaxillary line under local anesthesia and fluoroscopic guidance. An 8F bioptome was introduced through it, and biopsy specimens were taken from several sites on the costal and diaphragmatic pleura. After biopsy, PE was completely evacuated, and the sheath was removed. RESULTS: During the 2-year pilot study, we procured, on average, 14 adequate pleural specimens from each of the 28 patients (age range, 15 to 81 years) on the first attempt. Histopathologic examination revealed tuberculous pleuritis (18 patients), metastatic adenocarcinoma (1 patient), and nonspecific pleuritis (9 patients). Postprocedure, 25 patients had rapid symptomatic improvement without recurrence of PE. No major complications occurred during or after the procedure (mean follow-up period, 2 years). CONCLUSIONS: Our new biopsy procedure can be performed easily, safely, and with increased diagnostic sensitivity and patient comfort. Unlike other biopsy techniques, it provides adequate multiple-site pleural biopsy specimens, in all cases, on the first attempt without any morbidity and mortality. It has a therapeutic potential to provide rapid symptomatic relief and treatment by pleurodesis. We recommend this procedure for patients whose conditions remain undiagnosed after undergoing needle biopsy or for those who cannot tolerate it, before considering more aggressive diagnostic interventions. This is the best alternative when thoracoscopy or thoracotomy are not available or when patients are at high risk for complications from them. PMID- 15136391 TI - Can a standardized acupuncture technique palliate disabling breathlessness: a single-blind, placebo-controlled crossover study. AB - STUDY OBJECTIVES: The management of disabling breathlessness is poor, and a standardized form of acupuncture has been reported as offering benefit. This study was designed to evaluate the efficacy of standardized acupuncture treatment. DESIGN: A single-blind, randomized, crossover study. SETTING: This study was carried out on a domiciliary basis in Southampton (UK). INTERVENTIONS: This study evaluated a standardized acupuncture technique vs an appropriately validated placebo/control (mock transcutaneous electrical nerve stimulation [TENS]) for disabling, nonmalignant breathlessness (largely COPD). The acupuncture was provided by an appropriately trained nurse acupuncturist. Each patient received six treatments in each phase of the study, with an intervening 2 week washout period. MEASUREMENTS AND RESULTS: The primary outcome was worst breathlessness (visual analog scale, 0 to 100 mm), with the sample size based on an 80% power to detect a 10-mm difference between treatment means. Secondary outcomes included the St George's respiratory questionnaire score and treatment credibility. RESULTS: Thirty-six patients were entered into the study (33 with COPD), and 24 patients completed both treatment phases. The primary outcome improved significantly during the course of the study, but there were no significant treatment differences between acupuncture and the placebo/control of mock TENS for either primary or secondary outcomes. The placebo was shown to be a credible control. There was no evidence of a carryover effect from the first to second phase of the study. CONCLUSION: This standardized acupuncture technique does not show specific efficacy in disabling nonmalignant breathlessness, but those entered into the study did experience clinically significant benefit from both treatments. PMID- 15136392 TI - A randomized controlled trial of an antibiotic discontinuation policy for clinically suspected ventilator-associated pneumonia. AB - OBJECTIVE: To evaluate an antibiotic discontinuation policy for clinically suspected ventilator-associated pneumonia (VAP). DESIGN: Prospective, randomized, controlled clinical trial. SETTING: A medical ICU from a university-affiliated urban teaching hospital. PATIENTS: Between April 2002 and July 2003, 290 patients completed the clinical trial. INTERVENTIONS: Patients were assigned to have the duration of antibiotic treatment for VAP determined by an antibiotic discontinuation policy (discontinuation group) or their treating physician teams (conventional group). RESULTS: Severity of illness using APACHE (acute physiology and chronic health evaluation) II score (22.8 +/- 9.0 vs 23.2 +/- 9.4, p = 0.683) [mean +/- SD] and the clinical pulmonary infection score (7.1 +/- 0.9 vs 7.2 +/- 0.9, p = 0.222) were similar for both patient groups. The duration of antibiotic treatment for VAP was statistically shorter among patients in the discontinuation group compared to patients in the conventional antibiotic management group (6.0 +/- 4.9 days vs 8.0 +/- 5.6 days, p = 0.001). The occurrence of a secondary episode of VAP was not statistically different between these two groups (17.3% vs 19.3%, p = 0.667). Hospital mortality (32.0% vs 37.1%, p = 0.357) and ICU length of stay (6.8 +/- 6.1 days vs 7.0 +/- 7.3 days, p = 0.798) were also statistically similar. CONCLUSIONS: The application of an antibiotic discontinuation policy for clinically suspected VAP was associated with a decrease in the overall duration of antibiotic treatment. These findings suggest that shorter courses of empiric antibiotic therapy for patients treated for clinically suspected VAP can be safely achieved. PMID- 15136393 TI - Intensive care in patients with HIV infection in the era of highly active antiretroviral therapy. AB - STUDY OBJECTIVES: The use of highly active antiretroviral therapy (HAART) has dramatically improved morbidity and mortality in patients with HIV infection. The types of critical illness and their outcomes in HIV-infected patients in recent years is unknown. DESIGN: We reviewed the medical records of all patients admitted to the Medical ICU of Beth Israel Medical Center, NY, from January to June 2001 and compared their characteristics with patients admitted to the same unit from November 1991 to October 1992. RESULTS: Of 441 admissions in the first half of 2001, 63 admissions (14%) were in 53 HIV-seropositive patients. There were 65 admissions to the Medical ICU during the 1-year period spanning 1991 to 1992. Compared with the earlier period, the 2001 patients were more likely to be black (52% vs 26%, respectively; p < 0.01) and injection drug users (75% vs 48%, respectively; p < 0.01), and were less likely to be white (11% vs 23%, respectively; difference not significant) and homosexual men (6% vs 26%, respectively; p < 0.01). In 2001, patients were less likely to be admitted with respiratory failure (22% vs 54%, respectively; p < 0.01) and with Pneumocystis jiroveci pneumonia (formerly referred to as Pneumocystis carinii) [3% vs 34%, respectively; p < 0.001], and were more likely to be admitted with non-HIV related diseases (67% vs 12%, respectively; p < 0.001). Overall survival was much higher in the later period (71% vs 49%, respectively; p < 0.01). CONCLUSIONS: In the era of HAART, more patients with HIV infection were admitted to the ICU over a 12-month period than were 10 years previously. Patients were more likely to be injection drug users and were more likely to be admitted to the ICU because of non-HIV-associated conditions. PMID- 15136394 TI - Emphysema and pneumothorax after percutaneous tracheostomy: case reports and an anatomic study. AB - STUDY OBJECTIVE: Part 1: To describe cases of emphysema (subcutaneous and/or mediastinal) and pneumothorax after percutaneous dilational tracheostomy (PDT) in a series of 326 patients, and to review the existing literature describing the incidence and possible mechanisms. Part 2: To analyze the potential mechanisms for the development of emphysema and pneumothorax in human cadaver models. DESIGN: A retrospective analysis of PDTs, in combination with an anatomic study in human cadavers. MATERIALS AND METHODS: Part 1: All ICU patients who underwent PDT between 1997 and 2002 were enrolled in the study. We analyzed the cases of emphysema and pneumothorax. Similar cases were retrieved from the literature and underwent a systematic review. Part 2: The relevant anatomic structures were studied. We simulated the clinical situation after PDT in a human pathologic study in order to induce subcutaneous emphysema and pneumothorax. MEASUREMENTS AND RESULTS: Part 1: Five cases of subcutaneous emphysema (1.5%) and two cases of pneumothorax (0.6%) are described. In the literature search, we found 41 cases of emphysema (1.4%) and 25 cases of pneumothorax (0.8%) in a total of 3,012 patients. Part 2: Subcutaneous emphysema could easily be induced in a human cadaver model by inflating air in the pretracheal tissues and after posterior tracheal wall laceration. Air leakage was also possible through a fenestrated cannula via the space between the inner nonfenestrated cannula and outer cannula and then through the fenestration. CONCLUSIONS: We conclude that one mechanism for the development of emphysema is an imperfect positioning of the fenestrated cannula, whereby the fenestration is extraluminal. For this reason, fenestrated cannulas should not be used immediately after placement of a PDT. Posterior tracheal wall laceration is another mechanism responsible for emphysema after PDT. After perforation of the posterior tracheal wall, the pleural space can be reached easily. This may result in a pneumothorax. PMID- 15136395 TI - Survival of patients with bronchiectasis after the first ICU stay for respiratory failure. AB - STUDY OBJECTIVES: Respiratory failure (RF) is a frequent cause of death among patients with bilateral bronchiectasis. An ICU admission is commonly required, and neither short-term or long-term outcomes have been studied. DESIGN: We performed a retrospective study over a 10-year period (January 1990 to March 2000). All patients with bilateral bronchiectasis admitted for the first time in the medical ICU for RF were reviewed. Patients with cystic fibrosis were excluded. MEASUREMENTS AND RESULTS: Forty-eight patients (mean age +/- SD, 63 +/- 11 years; mean simplified acute physiology score [SAPS] II, 32 +/- 12) of whom 25% received long-term oxygen therapy (LTOT) were identified. All the patients were treated with intensive medical care, associated with noninvasive ventilation in 13 patients (27%), and 26 patients (54%) required intubation. Nine patients (19%) died in the ICU. The 1-year mortality rate was 40%. Among the variables recorded at ICU admission, age > 65 years (p = 0.002), SAPS II score > 32 (p = 0.012), use of LTOT (p = 0.047), and intubation (p = 0.027) were associated with reduced survival in univariate analysis by Cox regression. Multivariate analysis by Cox proportional hazard model showed that age > 65 years (relative risk [RR], 2.70; 95% confidence interval [CI], 1.15 to 6.29) and use of LTOT (RR, 2.52; 95% CI, 1.15 to 5.54) were independently associated with reduced survival. CONCLUSIONS: We performed the first study providing information related to the impact of the first ICU stay for RF on long-term outcomes for patients with bilateral bronchiectasis. Age > 65 years and prior use of LTOT were associated with reduced survival. PMID- 15136396 TI - Influence of i.v. haloperidol on ventricular repolarization and monophasic action potential duration in anesthetized dogs. AB - INTRODUCTION: i.v. haloperidol is used commonly for sedation in critically ill patients. However, i.v. haloperidol has been shown to cause the life-threatening ventricular tachyarrhythmia torsades de pointes. Mechanisms by which haloperidol causes torsades de pointes have not been widely investigated in controlled studies. STUDY OBJECTIVES: To determine the effects of i.v. haloperidol on electrophysiologic parameters known to promote torsades de pointes. INTERVENTIONS: Monophasic action potential catheters were guided under fluoroscopy into the right and left ventricles of 14 chloralose-anesthetized dogs (haloperidol, nine dogs; placebo, five dogs). Effective refractory period (ERP), action potential duration at 90% repolarization (APD90), and QTc interval measurements were performed at baseline and after each of four doses of haloperidol (0.15, 0.5, 2.0, and 3.0 mg/kg) or placebo at three different pacing cycle lengths (450, 300, and 250 ms). MEASUREMENTS AND RESULTS: i.v. haloperidol significantly prolonged left and right ventricular ERP by a magnitude of 12 to 20% at all pacing cycle lengths. ERP values in the placebo group did not change significantly from pretreatment values in either ventricle. Haloperidol significantly prolonged left ventricular APD90 at a pacing cycle length of 300 ms. The effects of haloperidol on right ventricular APD90 approached significance at a cycle length of 450 ms. Overall, haloperidol prolonged APD90 by 7 to 11%, with less consistent and more variable effects than those for the ERP. APD90 was not significantly altered in the placebo groups. Haloperidol produced significant prolongation in QTc intervals. The electrophysiologic effects of haloperidol were related to dose, with a plateau reached at the 0.5 mg/kg dose for ERP measurements and at the 2 mg/kg dose for the APD90 and QTc interval measurements. CONCLUSIONS: i.v. haloperidol prolongs ventricular ERP and APD90 in intact canine hearts. These electrophysiologic effects are likely associated with the clinical torsades de pointes-inducing actions of i.v. haloperidol in critically ill patients. PMID- 15136397 TI - Use of novel nonfluoroscopic three-dimensional electroanatomic mapping system to monitor and analyze heart surgery in animal models. AB - BACKGROUND: The new method of three-dimensional (3D) electroanatomic mapping was presented as an important tool for cardiac imaging and intervention. We present herein the first use of this technology for the monitoring, analysis, and development of cardiac surgery at the preclinical stage. METHODS: The method is based on utilizing a locatable catheter connected to an endocardial mapping and navigating system, to accurately establish the location and orientation of the tip of the mapping catheter and simultaneously record its local electrogram. The 3D geometry of the beating cardiac chamber is reconstructed in real time. The system was tested on six goats that underwent dynamic cardiomyoplasty. Two maps of each animal were performed: preoperative and postoperative during the stimulation protocol of the skeletal muscle. RESULTS: The electroanatomic mapping system provided detailed maps of the left ventricle during the stimulation protocol, which demonstrated a striking geometric difference between the assisted and the unassisted beats. These geometric changes are best described by referring to left ventricular long-axis movements (22.3 +/- 3.8 degrees vs 3.4 +/- 1.6 degrees, p < 0.001), center-of-mass movements (10.4 +/- 3.0 mm vs 3.9 +/- 1.6 mm, p < 0.005), and the changes in upward movement viewed along the base (7.9 +/- 1.9 mm vs 3.6 +/- 1.7 mm, p < 0.01), middle (13.8 +/- 4.0 mm vs 7.3 +/- 1.8 mm, p < 0.005), and the apex of the heart (28.1 +/- 4.5 vs 5.3 +/- 2.3 mm, p < 0.001) [mean +/- SD]. CONCLUSIONS: The 3D electroanatomic mapping system allows detailed reconstruction of the left ventricular geometry and a clear view of the difference between the assisted and the unassisted beats. This novel monitoring system may serve as an important tool for the analysis and development of new techniques in cardiac surgery. PMID- 15136398 TI - Expression of adhesion molecules during apoptosis of circulating neutrophils in COPD. AB - STUDY OBJECTIVES: Neutrophil accumulation occurs in the lungs of patients with COPD. This can be due to increased recruitment and/or delayed tissue clearance. Previous studies have described alterations in circulating neutrophils in these patients that can facilitate the former. Dysregulation of neutrophil apoptosis may contribute to the latter. This study investigated the potential abnormalities of the apoptotic process in COPD patients. DESIGN: Prospective study. SETTINGS: Outpatient clinic in a urban, tertiary hospital. PATIENTS: Fourteen stable patients with COPD, 8 smokers with normal lung function, and 8 healthy nonsmoking subjects. MEASUREMENTS AND RESULTS: We cultured circulating neutrophils that had been harvested from the study subjects at 2, 6, and 24 h. Apoptosis was assessed using flow cytometry by annexin binding and CD16 expression. The surface expression of the adhesion molecules Mac-1 (CD11b) and L-selectin (CD62L) also was determined by flow cytometry. The percentage of apoptotic neutrophils increased with time similarly in all groups. However, the surface expression of Mac-1 (CD11b) was higher, and that of L-selectin (CD62L) was lower, during apoptosis in the neutrophils of patients with COPD. CONCLUSIONS: These results show that, quantitatively, in vitro neutrophil apoptosis in COPD patients occurred at a rate similar to that found in healthy individuals and smokers with normal lung function. Qualitatively, however, the increased surface expression of Mac-1 (CD11b) and the decreased surface expression of L-selectin (CD62L) observed in the apoptotic neutrophils of COPD patients indicate increased activation during the apoptotic process. This may be relevant for the pathogenesis of COPD. PMID- 15136399 TI - Gene expression profiling identifies matriptase overexpression in malignant mesothelioma. AB - STUDY OBJECTIVE: We investigated the gene expression profiles of malignant pleural mesothelioma (MPM) specimens to identify novel genes that are potentially involved in the oncogenic transformation of human pleural cells. DESIGN: Complementary DNA (cDNA) microarray transcriptional profiling studies of 10 MPM cell lines and 4 MPM primary tumor specimens were performed using hierarchic clustering. To confirm microarray data, we used real-time polymerase chain reaction and immunoblotting. RESULTS: Cluster analysis differentiated among epithelial (E), sarcomatoid, and biphasic MPM variants. Expression profiling identified common overexpressed or underexpressed genes in MPM. Notably, matriptase messenger RNA was found to be overexpressed by 826-fold in E MPM, with protein expression subsequently confirmed by immunoblot analysis. This recently characterized trypsin-like serine protease has been implicated in tumor invasion and metastasis of E-derived cancers, but has not been described until now in MPM. We also identified other novel genes, such as insulin-like growth factor binding protein 5 and a cDNA clone similar to proteolipid MAL2. CONCLUSIONS: Thus, further large-scale profiling of MPM may elucidate previously unrecognized molecular mechanisms by identifying novel genes that are involved in malignant transformation. Our study has now found matriptase to be one of these mesothelioma-associated genes, with potential pathogenic and therapeutic significance. PMID- 15136400 TI - Native matrix metalloproteinase characteristics may influence early stenosis of venous versus arterial coronary artery bypass grafting conduits. AB - PURPOSE: Stenosis and occlusion rates of internal mammary artery (IMA) and saphenous vein (SV) coronary artery bypass grafts (CABGs) are markedly different, which result from respective disparities in vascular remodeling. Matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) regulate vascular structure and may have important influence on graft patency. However, the MMP milieu and expression profile of the IMA and SV have not been contrasted. Therefore, the aim of this study was to assess and compare the native MMP systems in IMA vs SV conduits. METHODS: IMA (n = 10) and SV (n = 10) specimens were obtained from patients undergoing CABG surgery. Protein levels of MMP-1, MMP-2, and MMP-9, TIMP-1, a membrane-bound MMP activator (MT1-MMP), and an extracellular MMP inducer protein (EMMPRIN) were determined by immunoblotting and quantified by densitometric analysis. MMP-2 and MMP-9 activity was determined by gelatin zymography. RESULTS: MMP-2 levels were significantly higher in SV (2,218 +/- 351 pixels) vs IMA (1,012 +/- 213 pixels) specimens (mean +/- SEM]). There were no significant differences in MMP-1, MMP-9, or TIMP-1 content; however, MT1 MMP and EMMPRIN levels were significantly lower in SV (847 +/- 190 pixels, 1,742 +/- 461 pixels) vs IMA conduits (2,590 + 403 pixels, 5,606 + 678 pixels), respectively (p < 0.05). MMP-9 activity was similar while MMP-2 activity was significantly increased in SV vs IMA specimens. CONCLUSIONS: SV and IMA conduits harbor the same MMP molecular constituents. However, MMP-2 levels and activity are significantly more abundant in the SV compared to the IMA. These differences may contribute to the early pathologic remodeling of the SV vs IMA conduit following CABG surgery. PMID- 15136401 TI - Recognizing and treating glucocorticoid-induced osteoporosis in patients with pulmonary diseases. AB - Glucocorticoids are frequently used to treat patients with pulmonary diseases, but continuous long-term use of glucocorticoids may lead to significant bone loss and an increased risk of fragility fractures. Patients with certain lung diseases, regardless of pharmacotherapy-particularly COPD and cystic fibrosis-and patients waiting for lung transplantation are also at increased risk of osteoporosis. Fragility fractures, especially of the hip, will have substantial effects on the health and well-being of older patients. Vertebral collapse and kyphosis secondary to glucocorticoid-induced osteoporosis (GIO) may affect lung function. Identification of patients with osteopenia, osteoporosis, or fragility fractures related to osteoporosis is strongly recommended and should lead to appropriate treatment. Prevention of GIO in patients receiving continuous oral glucocorticoids is also recommended. In patients receiving either high-dose inhaled glucocorticoids or low- to medium-dose inhaled glucocorticoids with frequent courses of oral glucocorticoids, bone mineral density measurements should be performed to screen for osteopenia and osteoporosis. A bisphosphonate (risedronate or alendronate), calcium and vitamin D supplementation, and lifestyle modifications are recommended for the prevention and treatment of GIO. PMID- 15136402 TI - Common causes of troponin elevations in the absence of acute myocardial infarction: incidence and clinical significance. AB - Cardiac troponin is a preferred biomarker of acute myocardial infarction (MI). Unfortunately, elevation of troponin can be detected in a variety of conditions other than acute MI. This review focuses on the incidence and clinical significance of increased troponin in conditions commonly associated with troponin elevations, particularly in those that may resemble acute MI. PMID- 15136404 TI - Guidelines for empiric antimicrobial prescribing in community-acquired pneumonia. AB - Empiric antimicrobial prescribing for community-acquired pneumonia remains a challenge, despite the availability of treatment guidelines. A number of key differences exist between North American and European guidelines, mainly in the outpatient setting. The North American approach is to use initial antimicrobial therapy, which provides coverage for Streptococcus pneumoniae plus atypical pathogens. Europeans tend to focus on providing pneumococcal coverage with less emphasis on covering for an atypical pathogen. Ambulatory patients without comorbidity are more likely to receive macrolide therapy in North America, whereas in Europe these patients would probably receive a beta-lactam agent. Major issues that are fundamental to this difference include the importance of providing therapy for atypical pathogens and the clinical significance of macrolide-resistant S pneumoniae. Prospective data are required to evaluate which of these two approaches offers clinical superiority. PMID- 15136405 TI - Is my lung function really that good? Flow-type spirometer problems that elevate test results. AB - Most spirometry errors reduce test results, and it is widely assumed that measurement accuracy is guaranteed by frequent spirometer calibrations or calibration checks. However, zero errors and changes in flow-type spirometer sensors may occur during testing that significantly elevate test results, even though the spirometer was calibrated recently. To draw attention to these often unrecognized problems, this report presents anomalous spirograms and test results obtained from occupational medicine clinics and hospital pulmonary function laboratories during quality assurance spirogram reviews. The spurious results appear to have been caused by inaccurate zeroing of the flow sensor, or by condensation, mucus deposition, or unstable calibration of various flow-type spirometers. These errors elevated some FVCs to 144 to 204% of predicted and probably caused 40% of 121 middle-aged working men in respirator medical clearance programs to record both FVC and FEV1 > 120% of predicted. Since spirometers report the largest values from a test, these errors must be recognized and deleted to avoid false-negative interpretations. Flow-type spirometer users at all levels, from the technician to the interpreter of test results, should be aware of the potential for and the appearance of these errors in spirograms. PMID- 15136406 TI - Patient-focused care: the 2003 American College of Chest Physicians Convocation Speech. PMID- 15136407 TI - Empiric therapy of community-acquired pneumonia: guidelines for the perplexed? AB - This article discusses the key clinical aspects of empiric therapy of community acquired pneumonia (CAP). Antibiotic selection, severity of CAP, single vs multiple pathogens, pharmacokinetic considerations, antibiotic resistance, i.v. vs oral antibiotic therapy for CAP, oral therapy for non-ICU hospitalized patients with CAP, beta-lactams, macrolides, ketolides, doxycycline, respiratory quinolones, and pharmacoeconomic implications are discussed. PMID- 15136408 TI - Effect of age and end point on the prognostic value of the exercise test. AB - BACKGROUND: The clinical and exercise test variables chosen for predicting prognosis vary in the available studies. This could be due to the effect of age of the patients tested and the choice of outcomes used as end points in these follow-up studies. OBJECTIVE: To evaluate the effect of age and end points on exercise test variables chosen as significantly and independently associated with time to death. METHODS: Analyses were performed on the first treadmill test performed on consecutive male veterans at the Palo Alto and Long Beach Veterans Affairs Medical Centers since 1987. After removal of patients with congestive heart failure, coronary interventions, left bundle-branch block, atrial fibrillation, myocardial infarction and/or Q wave, and digoxin use, 3,745 male subjects remained. The outcomes were cardiovascular and all-cause mortality. The study population was divided into subsets according to age; exercise test and clinical variables were analyzed within the age subsets using the Cox hazard model. RESULTS: The mean age at the time of testing was 57 +/- 12 years (+/- SD) and they were followed up for a mean of 6.6 years. There were 544 all-cause deaths, with 206 of the deaths being due to cardiovascular causes (38%). When the study group was classified into subsets based on age, exercise capacity (in metabolic equivalents [METs]) was chosen by the Cox hazard model most consistently in the age groups using either end point. Even when age was added to the Duke treadmill score, prediction of death did not improve in those > 70 years of age because of the nonlinear relationship between age, the exercise test variables, and time to death. The most important age cut points for clinically important differences in exercise test predictors appeared to be 70 years and 75 years of age. In the patients 70 to 75 years of age, peak METs was the only variable predictive of all-cause mortality, and exercise-induced ST-segment depression was the only predictor of cardiovascular death; in the patients > 75 years of age, none of the exercise test responses were predictive of either death outcome. CONCLUSION: Both age and the outcome selected as an end point affect the exercise test responses chosen for scores to predict prognosis. Differences in age of the subjects tested and/or the outcome selected as the end point can explain the differences in the studies using exercise testing to predict prognosis. PMID- 15136409 TI - Genetics of COPD. AB - COPD is a complex mix of signs and symptoms in patients with chronic bronchitis and emphysema, diseases that largely result from cigarette smoking. Not all smokers, however, acquire COPD, and COPD can develop in nonsmokers. In the United States, COPD is currently the fourth leading cause of death. Surprisingly, there are no effective drug therapies for COPD that are able to significantly alter disease progression, and little is known of the underlying molecular mechanisms that are responsible for its occurrence. Candidate gene-association studies and linkage analyses have been reported for COPD patients. This review describes the genetic predisposition of healthy subjects or relatives of COPD patients to acquire COPD. In addition, the genetic bases of COPD with rapid decline of FEV1 are described, and the current genetic data that have been distilled from studies of COPD patients with a predominant emphysema phenotype, with chronic bronchitis phenotype, and with a response to bronchodilators are discussed. PMID- 15136410 TI - A 67-year-old woman with abnormal lung perfusion scan finding. PMID- 15136411 TI - Hemoptysis in a 38-year-old woman receiving an oral contraceptive. PMID- 15136412 TI - Survival after prolonged submersion in freshwater in Florida. AB - A 2-year-old boy was submerged for at least 20 min in a freshwater creek in Union County, FL. Cardiopulmonary resuscitation (CPR) was administered for approximately 1 h at the scene and during transport to the hospital. On arrival, his Glasgow coma scale score was 3 and rectal temperature was < or = 26.7 degrees C. He demonstrated respiratory failure, intense vasoconstriction, hemoglobinuria, anemia, hypercoagulability, thrombocytosis, leukopenia, and persistent coma. With intensive care, he began emerging from the coma after 72 h and progressively improved. Testing at the Developmental Evaluation Center and clinical observations showed him to be completely normal by 6 months after drowning. Thus, severe, rapid hypothermia can occur during drowning in cold water in any geographic location and at temperatures above those necessary for ice formation. Hypothermia provides cerebral protection from hypoxia, permitting total recovery with appropriate CPR and intensive care. PMID- 15136413 TI - Kapalabhati pranayama: breath of fire or cause of pneumothorax? A case report. AB - Spontaneous pneumothorax is the most common cause of pneumothorax. We report a case of a 29-year-old healthy woman who presented to the emergency department with a spontaneous pneumothorax caused by a yoga breathing technique called Kapalabhati pranayama, or breath of fire. Yoga breathing exercises are commonly practiced, and a limited number of studies have shown various physiologic benefits of yoga breathing. This is the only known report of spontaneous pneumothorax caused by pranayama, but some other rare causes are noted. This case should illustrate that adverse side effects can occur when one pushes the body to physiologic extremes. PMID- 15136414 TI - Z alpha1-antitrypsin polymerizes in the lung and acts as a neutrophil chemoattractant. AB - BACKGROUND: Alpha1-antitrypsin (A1AT) is an abundant protein that is synthesized in the liver and is secreted into the plasma. From the plasma, A1AT diffuses into various body compartments, including the lung where it provides much of the antiprotease protection. The current understanding of the pathogenesis of emphysema in A1AT-deficient individuals focuses on the polymerization of mutant protein within the liver, which results in a deficiency of circulating A1AT and a protease-antiprotease imbalance in the lungs. METHODS AND RESULTS: In this study, we evaluated BAL fluid samples from five healthy volunteers, five individuals with ZA1AT deficiency, and an individual with the PiZZ phenotype who had received a liver transplant. We show that the lung itself is a source of A1AT. In addition, the Z protein formed in the lung polymerizes, and these polymers are detectable in lung epithelial lining fluid by enzyme-linked immunosorbent assay and Western blot analysis. Finally, we show that polymeric ZA1AT is a potent neutrophil chemoattractant that is similar to polymerized MA1AT. CONCLUSIONS: Our findings suggest that the polymerization of locally produced ZA1AT is a contributory factor to the lung inflammation experienced by those with A1AT deficiency and that standard antiprotease therapies may not address this problem. PMID- 15136415 TI - COPD and smoking cessation motivation. PMID- 15136416 TI - Early mobilization in pneumonia. PMID- 15136417 TI - Diagnosing neuroleptic malignant syndrome. PMID- 15136418 TI - Treatment with inhaled flunisolide. PMID- 15136419 TI - Plasma orexin-A levels in obstructive sleep apnea-hypopnea syndrome. PMID- 15136420 TI - Increased plasma interleukin-6 is associated with the pathogenesis of obstructive sleep apnea syndrome. PMID- 15136421 TI - Effect of repeated thoracenteses on fibrinolytic activity in malignant pleural effusion. PMID- 15136422 TI - Preoperative pulmonary consultation at a university hospital. PMID- 15136425 TI - Proceedings of the 46th Annual Thomas I Petty Lung Conference. Lung cancer: early events, early interventions. PMID- 15136426 TI - Environmental causes of lung cancer: what do we know in 2003? AB - The environmental causes of lung cancer have been the focus of intense epidemiologic and other research for > 50 years. The resulting evidence causally associates lung cancer with active and passive smoking, a variety of occupational agents, and indoor and outdoor air pollution. These causal associations have motivated control initiatives through education, regulation, and litigation. In recent years, the research focus has shifted to identifying the determinants of susceptibility to these agents, including interactions among environmental factors and genetic determinants of susceptibility to these agents. This article provides an overview of past and current research on the environment and lung cancer, and addresses the use of scientific evidence in controlling this cancer, which is largely caused by the environment. PMID- 15136427 TI - Increased urinary 8-isoprostaglandin F(2)alpha is associated with lower plasma selenium levels and lower vegetable and fruit intake in an asbestos-exposed cohort at risk for lung cancer. PMID- 15136428 TI - p53: at the crossroads of molecular carcinogenesis and molecular epidemiology. PMID- 15136429 TI - Development of lung tumors in mutant p53-expressing mice after inhalation exposure to asbestos. PMID- 15136430 TI - Genetic predisposition to lung cancer. PMID- 15136431 TI - Lung cancer risk in relatives of early-onset lung cancer cases. PMID- 15136432 TI - Premalignant evolution of lung cancer: Gilles F Filley lecture. PMID- 15136433 TI - Preinvasive bronchial lesions: surveillance or intervention? PMID- 15136434 TI - Molecular targets for cancer therapy and prevention. AB - Despite major improvements in patient management, the prognosis for patients with lung cancer remains dismal. As our knowledge of the molecular biology of cancers has increased, new targets for therapeutic interventions have been identified. In this article, we discuss some of the more recent developments in this field. They include revisiting some of the established concepts, such as retinoid metabolism and the inhibition of cyclooxygenase-2 metabolism. In addition, newer targets, such as transforming growth factor-beta signaling, Janus-activated kinase/signal transducers and activators of transcription pathway, and cell invasion are discussed. These studies demonstrate that multiple, often overlapping, mechanisms of disruption are present in lung cancer cells, presenting a plethora of molecular targets. PMID- 15136435 TI - Serial analysis of gene expression profiles of developmental stages in non-small cell lung carcinoma. PMID- 15136436 TI - Tobacco carcinogen-induced cellular transformation increases Akt activation in vitro and in vivo. PMID- 15136437 TI - Role of p63 amplification and overexpression in lung cancer development. PMID- 15136438 TI - Evaluation of H37, a candidate 3p21.3 tumor suppressor gene, as a therapeutic and diagnostic marker in lung cancer. PMID- 15136439 TI - Molecular classification of lung cancer: a cross-platform comparison of gene expression data sets. PMID- 15136440 TI - Class prediction of lung nodule gene expression profiles. PMID- 15136441 TI - Construction and optimization of chromosome arm-specific comparative genomic hybridization arrays for identifying genetic alterations in preinvasive lung cancers. PMID- 15136442 TI - Efficient identification and use of tumor-associated antibodies as markers of non small cell lung cancer. PMID- 15136443 TI - Caveolin-1 is down-regulated in human lung carcinoma and acts as a candidate tumor suppressor gene. PMID- 15136444 TI - Retrovirus-induced lung cancer: mechanisms of transformation of alveolar type II epithelial cells. PMID- 15136445 TI - Selective suppression of dendritic cell functions by cigarette smoke extract. PMID- 15136446 TI - Buccal-lung comparison of quantitative expression of carcinogen and oxidant metabolism genes in human subjects. PMID- 15136447 TI - Diffuse idiopathic pulmonary neuroendocrine cell hyperplasia as a precursor to pulmonary neuroendocrine tumors. PMID- 15136448 TI - High prevalence of endobronchial malignancy in high-risk patients with moderate dysplasia in sputum. PMID- 15136449 TI - Serum levels of surfactant protein D are increased in mice with lung tumors. PMID- 15136450 TI - Improvement and application of fluorescence inter-simple sequence repeat polymerase chain reaction for the study of subclonal growths in lung epithelial cell populations. PMID- 15136451 TI - Regulated gap junction-cytoskeletal associations in rat alveolar epithelial cells. PMID- 15136452 TI - Gene expression patterns, prognostic and diagnostic markers, and lung cancer biology. PMID- 15136453 TI - Sprouty 2 gene in mouse lung tumorigenesis. PMID- 15136454 TI - Non-small cell lung cancer molecular profiles are associated with lung development. PMID- 15136455 TI - Impact of cigarette smoke on the normal airway transcriptome. PMID- 15136456 TI - Elevated hepatocyte growth factor level correlates with poor outcome in early stage and late-stage adenocarcinoma of the lung. PMID- 15136457 TI - Epigenetics in lung cancer: focus on progression and early lesions. PMID- 15136458 TI - Gamma-catenin expression is reduced or absent in a subset of human non-small cell lung cancers, and its re-expression inhibits cell growth. PMID- 15136459 TI - Human lung cancer chemoprevention strategies: Parker B. Francis lecture. AB - Pharmacologic or nutritional prevention of lung cancers is needed, especially for 60 million Americans who are former smokers. A portfolio of large-scale trials of beta-carotene, beta-carotene with and without vitamin E, and beta-carotene plus vitamin A demonstrated no benefit whatsoever from beta-carotene. The alpha Tocopherol/beta-Carotene Trial and the beta-Carotene and Retinol Efficacy Trial found significant increases in lung cancer risk and total mortality. Laboratory research soon identified multiple adverse molecular effects. Nevertheless, chemoprevention remains an active, promising strategy, with new hypotheses and new candidate agents, including many already approved as therapies. The most active area currently is focused on selective inhibition of arachidonic metabolism, both Cox-2 and Lox pathways. PMID- 15136460 TI - Airway antioxidants and oxidative stress as predictors of sputum atypia and airflow obstruction. PMID- 15136461 TI - Chemoprevention of tobacco smoke-induced lung cancer: what animal studies tell us. PMID- 15136462 TI - Mechanisms of chemoprevention. PMID- 15136463 TI - The chemokine receptor, CXCR2, mediates the tumorigenic effects of ELR+ CXC chemokines. PMID- 15136464 TI - Ligands of peroxisome proliferator-activated receptor gamma inhibit lung cancer cell growth and induce apoptosis by stimulation of P21 expression. PMID- 15136465 TI - Cyclooxygenase-2 in lung carcinogenesis and chemoprevention: Roger S. Mitchell lecture. PMID- 15136466 TI - Cyclooxygenase-2-dependent expression of survivin in non-small cell lung cancer. PMID- 15136467 TI - Prostacyclin in human non-small cell lung cancers. PMID- 15136468 TI - Role of protein kinase B-dependent signaling in lung tumorigenesis. AB - Recent studies have defined the survival pathways activated by receptor tyrosine kinases that are critical in the transformation of human bronchial epithelial cells and in maintaining the survival of non-small cell lung cancer (NSCLC) cells. Protein kinase B (AKT) is one element of receptor tyrosine kinase signaling that is activated in bronchial premalignancy and NSCLC. Recent studies have shown that AKT cooperates with the stress kinase mitogen-activated protein kinase kinase 4 to maintain the survival of NSCLCs. These studies illustrate the importance of understanding the interactions between survival pathways and developing inhibitors to specific kinases that can be used alone or in combination in clinical trials for lung cancer prevention and treatment. PMID- 15136469 TI - Chemoprevention of lung cancer in transgenic mice. PMID- 15136470 TI - A multicenter study comparing autofluorescence bronchoscopy to white light bronchoscopy using a non-laser light stimulation system. PMID- 15136471 TI - New lung cancer drugs from bradykinin antagonists. PMID- 15136472 TI - Mechanisms of lung cancer chemoprevention by D-glucarate. PMID- 15136473 TI - Modulation of biomarkers during chemoprevention of mouse lung tumorigenesis by D glucarate. PMID- 15136474 TI - Potential role for activated leukocyte cell adhesion molecule and neural cadherin in metastasis to the lung microcirculation. PMID- 15136475 TI - Role of parathyroid hormone-related protein in lung cancer cell survival. PMID- 15136476 TI - Sputum dysplasia, related to bacterial load, oxidative stress, and airway inflammation in COPD. PMID- 15136477 TI - Lung cancer screening using low-dose high-resolution CT scanning in a high-risk workforce: 3500 nuclear fuel workers in three US states. PMID- 15136478 TI - Phospho-Akt overexpression in non-small cell lung cancer confers significant stage-independent survival disadvantage. PMID- 15136479 TI - Activation of the Akt/nuclear factor-kappaB signaling axis in developing lung neoplasia. PMID- 15136480 TI - Evidence that inflammation encourages pulmonary adenocarcinoma formation in mice: clinical implications. PMID- 15136481 TI - New targets for detection and treatment of small cell lung cancer. PMID- 15136482 TI - Use of standardized reverse transcription-polymerase chain reaction and the standardized expression measurement center in multi-institutional trials to develop meaningful lung cancer classification based on molecular genetic criteria. PMID- 15136483 TI - The SDF-1/CXCL 12/CXCR4 biological axis in non-small cell lung cancer metastases. PMID- 15136484 TI - A pilot evaluation of micrometastases for the prediction of outcome in lung cancer. PMID- 15136485 TI - Effects of budesonide on overall 5-methylcytosine levels and specific methylation and messenger RNA expression of the insulin-like growth factor II gene in mouse lung tumors. PMID- 15136486 TI - Improved diagnostic sensitivity for lung cancer using an automated quantitative cytology system and uridine 5'-triphosphate-induced sputum specimens. PMID- 15136487 TI - Screening for lung cancer: current status and future directions: Thomas A. Neff lecture. AB - Lung cancer is the number one cancer killer in North America. Currently, screening for lung cancer is not recommended. Therefore, patients will not receive a diagnosis until they present with symptomatic disease, which is usually advanced stage disease. Previous trials of screening with chest roentgenograms and sputum cytology have failed to show a decrease in lung cancer mortality. Some reports of screening with low-dose spiral CT scans have detected lung cancers at a smaller size (average size, 1.5 cm) than those usually detected by chest radiographs (mean size, 3.0 cm). Spiral CT scanning has been shown to detect between 58% and 85% of non-small cell lung cancers (NSCLCs) while they are in stage IA, and this compares favorably to the current medical practice, in which only 15% are detected as localized disease (Surveillance, Epidemiology, and End Results study data). This article summarizes the spiral CT screening data, and reviews some of the data related to screening with sputum cytology, sputum methylation, and autofluorescence bronchoscopy. Last, there is a brief discussion of some promising future strategies, with emphasis and data from studies presented at this Aspen Lung Conference. PMID- 15136488 TI - Lung cancer detection in asymptomatic patients with airflow obstruction. PMID- 15136489 TI - Bimodality lung cancer screening in high-risk patients: a preliminary report. PMID- 15136490 TI - Detection of melanoma antigen-A expression in sputum and bronchial lavage fluid of patients with lung cancer. PMID- 15136491 TI - Experience with surgical intervention for screening-detected lung cancer. PMID- 15136492 TI - Lung cancer: early events, early interventions: conference summary for the 46th Annual Thomas L Petty Aspen Lung Conference. PMID- 15136493 TI - Soluble CD40 ligand, soluble P-selectin, interleukin-6, and tissue factor in diabetes mellitus: relationships to cardiovascular disease and risk factor intervention. AB - BACKGROUND: High levels of the soluble fragment of CD40 ligand (sCD40L) have previously been associated with adverse cardiovascular outcomes. CD40L-CD40 interaction has been linked to the pathogenesis of atherothrombotic complications in cardiovascular disease (CVD). We sought to determine whether a "package of care" of intensified multifactorial cardiovascular risk intervention could reduce indices of platelet activation, inflammation, and coagulation in diabetes and whether patients with overt CVD would derive similar benefit compared with those without. METHODS AND RESULTS: We measured plasma sCD40L, soluble P-selectin (sP sel, an index of platelet activation), interleukin-6 (IL-6, a proinflammatory cytokine), and tissue factor (TF, an initiator of coagulation) in 97 patients with diabetes mellitus (41 with and 56 without overt CVD) and 39 comparable healthy control subjects. Thirty-six patients with and 32 without overt CVD then participated in a package of care of cardiovascular risk intervention over a period of 1 year. Plasma levels of sCD40L (P<0.001), sP-sel (P<0.001), IL-6 (P=0.001), and TF (P<0.001) were higher in patients with diabetes than in control subjects, with TF levels highest in patients with overt CVD. Multifactorial intervention was associated with significant reductions in sCD40L in both patient groups (both P<0.001), but reductions in sP-sel and TF were seen only in patients without overt CVD. There was no significant change in IL-6 levels in both patient groups. CONCLUSIONS: Intensive multifactorial risk management can reduce high levels of sCD40L but can only partially correct abnormal platelet activation, inflammation, and coagulation in diabetes, particularly in patients with overt CVD. PMID- 15136494 TI - Left septal atrial flutter: electrophysiology, anatomy, and results of ablation. AB - BACKGROUND: We describe the clinical and electrophysiological characteristics of a novel macroreentrant form of left atrial flutter circuit. METHODS AND RESULTS: A total of 11 patients were included in the study. The mean tachycardia cycle length was 278+/-41 ms. Nine of the 11 patients were treated with antiarrhythmic drugs at the time of the study for concomitant atrial fibrillation. With the use of entrainment pacing and either the CARTO Biosense mapping system (9 patients) or conventional mapping (2 patients), the flutter circuit was found to rotate around the left septum primum with a critical isthmus located between the pulmonary veins posteriorly and/or mitral annulus anteriorly and the septum primum. In 5 patients, radiofrequency ablation was performed from the septum primum to the right inferior pulmonary vein (group 1), and in 6 patients, a lesion was made from the septum primum to the mitral annulus (group 2). After a follow-up of 13+/-6 months, 2 patients in group 1 and all patients in group 2 remained in sinus rhythm without recurrence. CONCLUSIONS: Slowing of electric conduction in the left atrial septum due to antiarrhythmic drugs and/or atrial myopathy seems to promote left septal atrial flutter. Radiofrequency ablation of this arrhythmia is usually effective and safe. A line of block between the septum primum and the mitral annulus proved to be effective for cure of tachycardia. PMID- 15136495 TI - Clinical recovery from end-stage heart failure using left-ventricular assist device and pharmacological therapy correlates with increased sarcoplasmic reticulum calcium content but not with regression of cellular hypertrophy. AB - BACKGROUND: Left ventricular assist device (LVAD) treatment is known to lead to structural and functional cellular modifications in the heart. The relevance of these changes for clinical recovery is unknown. METHODS AND RESULTS: We compared properties of cardiomyocytes obtained from tissue taken at explantation of the LVAD in patients with clinical recovery with those obtained from hearts of patients who did not show clinical recovery, thus requiring transplantation. Compared with myocytes taken at implantation, both the recovery and nonrecovery groups showed approximately 50% reduction in cell capacitance, an index of cell size. However, action potential duration shortened, L-type Ca2+ current fast inactivation was more rapid, and sarcoplasmic reticulum Ca2+ content was increased in the recovery compared with the nonrecovery group. CONCLUSIONS: These results show that specific changes in excitation-contraction coupling, and not regression of cellular hypertrophy, are specifically associated with clinical recovery after LVAD and further identify sarcoplasmic reticulum Ca2+ handling as a key functional determinant in patients with heart failure. PMID- 15136496 TI - Hypoplastic left heart syndrome with intact or highly restrictive atrial septum: outcome after neonatal transcatheter atrial septostomy. AB - BACKGROUND: Hypoplastic left heart syndrome (HLHS) with intact or very restrictive atrial septum is a highly lethal combination. We review our 13-year institutional experience treating this high-risk subgroup of patients with emergent catheter therapy. METHODS AND RESULTS: Infants with HLHS requiring catheter septostomy within the first 2 days of life were compared with a matched control group with adequate interatrial communication. Preoperative, early postoperative, and medium-term survival were evaluated. Earlier experience was compared with recent results to assess the effect of changes in catheterization and surgical and intensive care unit management strategies over the study period. From 1990 to 2002, 33 newborns with HLHS (11% of newborns with HLHS managed during this period) underwent urgent/semiurgent catheterization to create or enlarge an interatrial communication before surgical palliation. Preoperative and early postoperative mortality were high (48%) compared with control HLHS patients, regardless of prenatal diagnosis and despite successful catheter-based atrial septostomy with clinical stabilization. Mortality trended down during the later part of the study period. Those who survived the neonatal period had late survival, pulmonary artery pressure, and resistance similar to those of control subjects. CONCLUSIONS: Neonatal mortality in the subgroup of HLHS patients with intact or highly restrictive atrial septum remains high despite successful urgent septostomy. Persistently poor outcomes for these patients have prompted efforts at our center to develop techniques for fetal intervention for this condition, in the hope that prenatal relief of left atrial and pulmonary venous hypertension may promote normal pulmonary vascular and parenchymal development and improve both short- and long-term outcomes. PMID- 15136497 TI - Relaxin is an independent risk factor predicting death in male patients with end stage kidney disease. AB - BACKGROUND: Patients with end-stage kidney disease (ESKD) have a reduced life expectancy mainly as the result of cardiovascular diseases. Relaxin has been implicated in the pathogenesis of cardiovascular diseases. We analyzed the impact of relaxin on death in patients with ESKD. METHODS AND RESULTS: Patients (n =245; 122 women, 123 men) on long-term hemodialysis were followed for 1140 days for death. Blood samples for analysis of relaxin, C-reactive protein, Troponin T, cholesterol, HDL, brain natriuretic peptide, and albumin were taken at study entry. Survival was compared by the Kaplan-Meier method and Cox regression analysis. One hundred seven patients died during the observation period; 66 died of cardiovascular diseases and 28 died of infectious diseases. Elevated serum relaxin concentrations (greater than median) predicted death in male but not in female patients with ESKD: All-cause death (men: relative risk, 2.63; 95% CI, 1.34 to 5.12; P=0.005; women: relative risk, 0.671; 95% CI, 0.33 to 1.35; P=0.262) and cardiovascular death (men: relative risk, 2.95; 95% CI, 1.20 to 7.21; P=0.018; women: relative risk, 0.639; 95% CI, 0.26 to 1.56; P=0.324). CONCLUSIONS: Relaxin is an independent risk factor predicting death in male patients with ESKD on chronic hemodialysis. PMID- 15136498 TI - Endothelial function predicts future development of coronary artery disease: a study of women with chest pain and normal coronary angiograms. AB - BACKGROUND: The prognosis for women with chest pain and angiographically normal coronary arteries is believed to be totally benign. Previous studies, however, did not account for the delay of a decade or so in the development of coronary artery disease that women may experience. METHODS AND RESULTS: This study assessed long-term follow-up of 42 women with de novo angina, evidence of reversible myocardial perfusion defects on SPECT, and normal coronary angiograms. At recruitment, all women underwent endothelial function testing (intracoronary acetylcholine) during catheterization. Patients were followed up for >10 years. Angiography was repeated at the end of the follow-up in 37 patients. At recruitment, 22 patients developed diffuse vasoconstriction during acetylcholine in the absence of identifiable focal coronary spasm (acetylcholine-positive group). The remaining 20 patients showed vasodilation (acetylcholine-negative group). At the end of follow-up, in the acetylcholine-positive group, 1 patient developed cardiac death, 13 still complained of chest pain, and 8 had remission of symptoms. In the acetylcholine-negative group, all patients showed complete resolution of chest pain beginning 6 to 36 months after baseline assessment. Angiography showed development of coronary artery disease in the 13 symptomatic patients in the acetylcholine-positive group. CONCLUSIONS: In women with angiographically normal-appearing coronary arteries, persistence of chest pain over the years often relates to development of coronary artery disease. Endothelial dysfunction in a setting of normal coronary arteries is a sign of future development of atherosclerosis. PMID- 15136499 TI - Early morning attenuation of endothelial function in healthy humans. AB - BACKGROUND: Cardiovascular events such as myocardial infarction, sudden death, and stroke have a peak incidence in the early hours after waking. The mechanisms involved in this circadian variation are not clear. Endothelial dysfunction is associated with increased risk for cardiovascular events. We tested the hypothesis that endothelial function is reduced in the early morning, around the time of waking, compared with measurements obtained both before sleep and later in the day in healthy humans. METHODS AND RESULTS: We studied 30 subjects (19 men, 11 women; mean age, 41.6 years). All participants underwent polysomnography to exclude obstructive sleep apnea or other sleep disorders. Brachial artery flow mediated endothelium-dependent vasodilation (FMD) and endothelium-independent dilation (non-FMD) were measured on 3 different occasions: before subjects went to sleep (9 PM), the next morning immediately after waking (6 AM), and during the late morning 5 hours after waking (11 AM). All subjects had normal sleep with good sleep efficiency of 84+/-2%. Compared with before sleep, FMD decreased markedly in the early morning after waking and recovered by late morning (9 pm, 7.5+/-1%; 6 am, 4.4+/-0.7%; 11 am, 7.7+/-1%; P=0.02). Non-FMD was similar for the 3 periods of observation (9 pm, 17.3+/-1.6%; 6 am, 17.2+/-1.3%; 11 am, 18.5+/ 1.7%). CONCLUSIONS: FMD is blunted in the early morning in healthy subjects. Decreased endothelial function in the early morning may have implications for our understanding of the morning peak in cardiac and vascular events. PMID- 15136500 TI - Differential effects of green tea-derived catechin on developing versus established atherosclerosis in apolipoprotein E-null mice. AB - BACKGROUND: Oxidative stress has been implicated in vascular injury and atherogenesis, and antioxidant treatment has shown favorable results in preclinical studies. Despite this, antioxidant therapy has failed to show benefit in clinical trials. Failure of antioxidants in clinical trials may be partly because such therapy is started after atherosclerosis is already well established, whereas the benefits in animal models may be results from early initiation of antioxidants while atherosclerosis is still evolving. METHODS AND RESULTS: To test this hypothesis, we evaluated the effect of epigallocatechin gallate (EGCG), the main antioxidant derived from green tea, on evolving and established atherosclerotic lesions in hypercholesterolemic apolipoprotein E-null mice. Established native aortic atherosclerotic lesions and evolving atherosclerotic lesions produced by periadventitial cuff injury to carotid arteries were assessed in mice after 21 and 42 days of treatment with daily intraperitoneal injections of EGCG (10 mg/kg) or PBS. EGCG treatment resulted in an increase in the antioxidant capacity in local vascular tissue and systemic circulation and reduced vascular smooth muscle cell proliferation and redox sensitive gene activation in vitro. EGCG reduced cuff-induced evolving atherosclerotic plaque size at 21 and 42 days by 55% and 73%, respectively, compared with PBS treatment (P<0.05). Conversely, EGCG had no effect on established lesions in the aortic sinuses or the rest of the aorta. CONCLUSIONS: Our data suggest that antioxidant EGCG differentially reduces evolving atherosclerotic lesions without influencing established atherosclerosis in the apolipoprotein E-null mice. PMID- 15136501 TI - AKT participates in endothelial dysfunction in hypertension. AB - BACKGROUND: In hypertension, reduced nitric oxide production and blunted endothelial vasorelaxation are observed. It was recently reported that AKT phosphorylates and activates endothelial nitric oxide synthase and that impaired kinase activity may be involved in endothelial dysfunction. METHODS AND RESULTS: To identify the physiological role of the kinase in normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR), we used adenoviral vectors to transfer the human AKT1 gene selectively to the common carotid endothelium. In vitro, endothelial vasorelaxations to acetylcholine, isoproterenol, and insulin were blunted in control carotids from SHR compared with WKY rats, and human AKT1 overexpression corrected these responses. Similarly, blood flow assessed in vivo by Doppler ultrasound was reduced in SHR compared with WKY carotids and normalized after AKT1 gene transfer. In primary cultured endothelial cells, we evaluated AKT phosphorylation, activity, and compartmentalization and observed a mislocalization of the kinase in SHR. CONCLUSIONS: We conclude that AKT participates in the settings of endothelial dysfunction in SHR rats by impaired membrane localization. Our data suggest that AKT is involved in endothelium dysfunction in hypertension. PMID- 15136502 TI - Mechano-electrical interaction late after Fontan operation: relation between P wave duration and dispersion, right atrial size, and atrial arrhythmias. AB - BACKGROUND: The growing population with Fontan operation surviving into adulthood has significant morbidity and mortality rates from recurrent atrial tachyarrhythmias. We hypothesized that the structural characteristics and electrical behavior of atria may differ in these patients compared with those without arrhythmias. METHODS AND RESULTS: We studied 33 consecutive patients (age, 25.4+/-9.5 years) with Fontan circulation, of whom 19 had a history of documented sustained atrial tachyarrhythmias. We analyzed their clinical and investigational data, including echocardiographic assessment of atrial dimensions and surface 12-lead ECG measurement of the P-wave duration and its dispersion between leads. Twenty age- and sex-matched healthy control subjects were also studied. First, patients who had the Fontan procedure overall had longer P-wave duration (144+/-33 versus 100+/-7 ms, P<0.001) and greater P-wave dispersion (74+/-33 versus 34+/-9 ms, P<0.001) than control subjects. Among the patients who had the Fontan procedure, those with atrial tachyarrhythmias had longer P-wave duration (159+/-28 versus 123+/-28 ms, P<0.001) and greater P-wave dispersion (91+/-30 versus 50+/-19 ms, P<0.001) than those without. Second, the patients with atrial tachyarrhythmias who had the Fontan procedure had larger right atrial dimension than those without arrhythmias (6.4+/-1.4 versus 5.0+/-1.0 cm, P=0.01). Third, both P-wave duration and dispersion were significantly correlated to right atrial dimension within the Fontan group (r=0.55, P=0.002, and r=0.56, P=0.002, respectively). CONCLUSIONS: Patients with atrial tachyarrhythmias late after Fontan operation have longer P-wave duration and P-wave dispersion and larger right atrial dimension than those without the arrhythmias; these abnormalities are interrelated. This observation represents an atrial mechano-electrical remodeling phenomenon in parallel to an increase in arrhythmia propensity in this vulnerable population and warrants further investigation. PMID- 15136503 TI - Microvascular resistance is not influenced by epicardial coronary artery stenosis severity: experimental validation. AB - BACKGROUND: The effect of epicardial artery stenosis on myocardial microvascular resistance remains controversial. Recruitable collateral flow, which may affect resistance, was not incorporated into previous measurements. METHODS AND RESULTS: In an open-chest pig model, distal coronary pressure was measured with a pressure wire, and the apparent minimal microvascular resistance was calculated during peak hyperemia as pressure divided by flow, measured either with a flow probe around the coronary artery (R(micro app)) or with a novel thermodilution technique (apparent index of microcirculatory resistance [IMR(app)]). These apparent resistances were compared with the actual R(micro) and IMR after the coronary wedge pressure and collateral flow were incorporated into the calculation. Measurements were made at baseline (no stenosis) and after creation of moderate and severe epicardial artery stenoses. In 6 pigs, 189 measurements of R(micro) and IMR were made under the various epicardial artery conditions. Without consideration of collateral flow, R(micro app) (0.43+/-0.12 to 0.46+/ 0.10 to 0.51+/-0.11 mm Hg/mL per minute) and IMR(app) (14+/-4 to 17+/-7 to 20+/ 10 U) increased progressively and significantly with increasing epicardial artery stenosis (P<0.001 for both). With the incorporation of collateral flow, neither R(micro) nor IMR increased as a result of increasing epicardial artery stenosis. CONCLUSIONS: After collateral flow is taken into account, the minimum achievable microvascular resistance is not affected by increasing epicardial artery stenosis. PMID- 15136504 TI - The microsomal triglyceride transfer protein gene-493T variant lowers cholesterol but increases the risk of coronary heart disease. AB - BACKGROUND: The microsomal triglyceride transfer protein (MTP) transfers lipids into apolipoprotein B-containing lipoproteins for secretion from liver, intestine, and heart. The T-variant of a functional polymorphism in the MTP promoter, MTP-493G/T, has been associated with reduced low-density lipoprotein cholesterol concentrations. We hypothesize that this polymorphism impacts on coronary heart disease (CHD) risk. METHODS AND RESULTS: The effect of the polymorphism was therefore tested in the West of Scotland Coronary Prevention Study biobank (580 cases and 1160 controls). MTP-493T carrier status was associated with significantly increased risk of CHD despite a small reduction in total cholesterol. Compared with the genotypic group with the lowest event rate (MTP-493GG, pravastatin treatment), the respective odds ratios (95% confidence interval) in the placebo group for CHD events were: GG, 1.23 (0.92 to 1.63); GT, 1.53 (1.12 to 2.08); and TT, 2.78 (1.53 to 5.05), suggestive of a gene-dose effect. The excess risk for CHD of the MTP-493T-variant was eliminated by pravastatin treatment. The Uppsala Longitudinal Study of Adult Men (ULSAM), which is a 20-year follow-up study of CHD, was used as an independent confirmatory database. These unexpected findings prompted the investigation of non-plasma lipid factors that could associate the MTP gene with CHD risk. In a limited number of subjects (n=18), heart muscle biopsies showed a MTP-493T genotype specific depression of MTP mRNA expression. CONCLUSIONS: The MTP-493T variant confers an increased risk of CHD that is unrelated to plasma lipids and lipoproteins, but eliminated by pravastatin treatment. A direct effect of the MTP polymorphism on myocardial lipid metabolism and vulnerability upon ischemic damage cannot be excluded. PMID- 15136505 TI - Impaired microvascular function in obesity: implications for obesity-associated microangiopathy, hypertension, and insulin resistance. AB - BACKGROUND: Obesity is associated with an increased risk of developing microangiopathy, hypertension, and insulin resistance. We hypothesized that obesity is a primary cause of microvascular dysfunction, which may contribute to the development of these obesity-related disorders. METHODS AND RESULTS: We examined microvascular function in 16 lean (body mass index <24 kg/m2) and 12 obese (body mass index >30 kg/m2) healthy women (mean age, 38.9+/-6.7 years) in the basal state and during physiological systemic hyperinsulinemia. We determined skin capillary recruitment after arterial occlusion with capillaroscopy and skin endothelium-(in)dependent vasodilation by iontophoresis of acetylcholine and sodium nitroprusside. Obese women, compared with lean women, had higher systolic blood pressure (P<0.05), impaired insulin sensitivity (P<0.01), impaired capillary recruitment in the basal state (P<0.05) and during hyperinsulinemia (P<0.05), and impaired acetylcholine-mediated vasodilation in the basal state (P<0.05) and during hyperinsulinemia (P<0.01). Sodium nitroprusside-mediated vasodilation was similar in lean and obese women. Capillary recruitment and acetylcholine-mediated vasodilation were positively correlated with insulin sensitivity (r=0.58, P<0.01 and r=0.55, P<0.01, respectively) and negatively with blood pressure (r=-0.64, P<0.001 and r=-0.42, P<0.05, respectively) in both lean and obese women. CONCLUSIONS: Obesity is characterized by impaired microvascular function in the basal state and during hyperinsulinemia and, in both lean and obese women, microvascular dysfunction is associated with increased blood pressure and decreased insulin sensitivity. These findings are consistent with a contribution of impaired microvascular function to the development of obesity related microangiopathy, hypertension, and insulin resistance. PMID- 15136506 TI - Chronic hyperglycemia attenuates coronary collateral development and impairs proliferative properties of myocardial interstitial fluid by production of angiostatin. AB - BACKGROUND: Development of coronary collateral vessels is impaired in patients with diabetes mellitus. We tested the hypothesis that hyperglycemia alone attenuates collateral development and abolishes proliferative properties of myocardial interstitial fluid (MIF) by enhancing expression of matrix metalloproteinases (MMP) and angiostatin. METHODS AND RESULTS: Chronically instrumented dogs were randomly assigned to receive an infusion of normal saline (control; n=9) or 70% dextrose in water to increase blood glucose to 350 to 400 mg/dL for 8 h/d (hyperglycemia; n=7) in the presence or absence (sham; n=9) of brief (2 minutes), repetitive coronary artery occlusions (1/h; 8/d for 21 days). Collateral perfusion increased to 41+/-11% and 49+/-6% of normal zone flow in control dogs on days 14 and 21 (P<0.05) but remained unchanged over 21 days in hyperglycemic and sham dogs (12+/-3% and 13+/-3%, respectively). A progressive reduction of the postocclusive peak reactive hyperemic response was also observed in control dogs (16+/-1 to 10+/-1 Hz. 10(2) on days 1 and 21, respectively) but not in hyperglycemic (17+/-2 to 20+/-2) or sham (17+/-2 to 16+/-1) dogs. Endothelial cell tube formation was produced by MIF obtained from control dogs but not hyperglycemic or sham dogs. Coincubation of MIF from hyperglycemic dogs with an angiostatin antibody restored endothelial cell tube formation. MMP-9 activity and expression of angiostatin were increased in dogs receiving exogenous glucose compared with controls CONCLUSIONS: Chronic hyperglycemia abolishes development of coronary collateral vessels by increasing MMP-9 activity and angiostatin expression in dogs. PMID- 15136507 TI - C-reactive protein promotes monocyte chemoattractant protein-1--mediated chemotaxis through upregulating CC chemokine receptor 2 expression in human monocytes. AB - BACKGROUND: Inflammation plays a crucial role in atherosclerosis. An elevated serum C-reactive protein (CRP) level is a strong marker for future atherosclerotic cardiovascular diseases. In addition, recent data suggest that CRP may directly promote atherogenesis. In this study, we investigated whether CRP can directly activate human circulating monocytes. METHODS AND RESULTS: Incubation of THP-1 monocytes with CRP (10 microg/mL) increased CC chemokine receptor 2 (CCR2) expression at both the protein and transcript levels, which in turn enhanced chemotaxis mediated by monocyte chemoattractant protein-1 (MCP-1) up to 2-fold. The CRP-induced upregulation of CCR2 expression involved binding of CRP to the FcgammaR, most notably FcgammaRI, and phospholipase D1 activation. Serum high-sensitivity CRP levels in 52 normocholesterolemic human subjects were positively correlated with CCR2 surface expression on circulating monocytes (r=0.62, P<0.001) and MCP-1-mediated monocyte chemotaxis (r=0.53, P<0.001). CONCLUSIONS: Elevated blood CRP levels may promote accumulation of monocytes in the atherogenic arterial wall by increasing chemotactic activities of monocytes in response to MCP-1. PMID- 15136508 TI - How many grails do we need? PMID- 15136509 TI - Spiral computed tomography for acute pulmonary embolism. AB - There is still considerable debate about the optimal diagnostic imaging modality for acute pulmonary embolism. If imaging is deemed necessary from an initial clinical evaluation such as d-dimer testing, options include nuclear medicine scanning, catheter pulmonary angiography, and spiral CT. In many institutions, spiral CT is becoming established as the first-line imaging test in daily clinical practice. With spiral CT, thrombus is directly visualized, and both mediastinal and parenchymal structures are evaluated, which may provide important alternative or additional diagnoses. However, limitations for the accurate diagnosis of small peripheral emboli, with a reported miss rate of up to 30% with single-slice spiral CT so far, have prevented the unanimous embrace of spiral CT as the new standard of reference for imaging pulmonary embolism. The clinical significance of the detection and treatment of isolated peripheral pulmonary emboli is uncertain. Evidence is accumulating that it is safe practice to withhold anticoagulation in patients with suspected pulmonary embolism on the basis of a negative spiral CT study. Remaining concerns about the accuracy of spiral CT for pulmonary embolism detection may be overcome by the introduction of multidetector-row spiral CT. This widely available technology has improved visualization of peripheral pulmonary arteries and detection of small emboli. The most recent generation of multidetector-row spiral CT scanners appears to outperform competing imaging modalities for the accurate detection of central and peripheral pulmonary embolism. In this review, we assess the current role and future potential of CT in the diagnostic algorithm of acute pulmonary embolism. PMID- 15136510 TI - Images in cardiovascular medicine. Unique discordance: thoracic situs solitus with left isomerism. PMID- 15136511 TI - Images in cardiovascular medicine. Life-threatening neonatal arrhythmia: successful treatment and confirmation of clinically suspected extreme long QT syndrome-3. PMID- 15136512 TI - Coronary endothelial dysfunction and cerebrovascular events. PMID- 15136513 TI - Unambiguous identification of implanted cells after cellular cardiomyoplasty: a critical issue. PMID- 15136514 TI - Pregnancy-associated plasma protein a as predictor of outcome in patients with suspected acute coronary syndromes. PMID- 15136515 TI - Early intervention with atorvastatin modulates TH1/TH2 imbalance in patients with acute coronary syndrome: from bedside to bench. PMID- 15136516 TI - Body mass index is not a risk factor for unstable angina and myocardial infarction in veteran patients with angiographically confirmed coronary artery disease. PMID- 15136517 TI - Simple self-help maneuvers are effective in aborting vasovagal syncope. PMID- 15136518 TI - Sex hormones and membrane microviscosity in elderly women. PMID- 15136519 TI - Impact of time to treatment on mortality after prehospital fibrinolysis or primary angioplasty. PMID- 15136520 TI - Everolimus stent proves effective. PMID- 15136521 TI - Repairing the foundation: the future of primary care. PMID- 15136526 TI - Food fights: common good versus individual interests. PMID- 15136528 TI - Medical privacy is important. PMID- 15136529 TI - Genetics and ARMD. PMID- 15136530 TI - Universal care. PMID- 15136531 TI - Universal care. PMID- 15136533 TI - Universal care. PMID- 15136534 TI - Radiologists get that long distance feeling. PMID- 15136536 TI - Prime Minister revives home care, pharmacare. PMID- 15136537 TI - Amended patent legislation still flawed. PMID- 15136539 TI - Saskatchewan's new system tracks surgical waits. PMID- 15136540 TI - International research sheds light on waiting times. PMID- 15136541 TI - AIDS strategy funding. PMID- 15136542 TI - Natural products and adverse drug interactions. PMID- 15136543 TI - Hormone replacement therapy for the primary prevention of chronic diseases: recommendation statement from the Canadian Task Force on Preventive Health Care. PMID- 15136544 TI - Nutrition labelling tidbits. PMID- 15136545 TI - Does perioperative oral rofecoxib therapy improve functional recovery after knee replacement surgery? PMID- 15136546 TI - A 17-year-old male with an unusual case of peritonitis. PMID- 15136547 TI - Cervical cancer mortality by neighbourhood income in urban Canada from 1971 to 1996. AB - BACKGROUND: The reduction of socioeconomic inequalities in health is an explicit objective of health policy in Canada, yet rates of death from cervical cancer are known to be higher among women of low socioeconomic status than among those of higher socioeconomic status. To evaluate progress toward the World Health Organization's goal of "Health for All," we examined whether income-related differentials in cervical cancer mortality diminished from 1971 to 1996. METHODS: Death registration data for Canada's census metropolitan areas in 1971, 1986, 1991 and 1996 were assigned to census tracts through postal code, and the tracts were in turn assigned to income quintiles based on their proportion of the population below the Statistics Canada low-income cutoff values. We compared age standardized death rates (using the 1966 world population standard) in the female population (excluding those in institutions) across the 5 income quintiles and calculated interquintile rate ratios (poorest over richest) and interquintile rate differences (poorest minus richest). RESULTS: From 1971 to 1996, the overall age-standardized cervical cancer death rate per 100 000 women (and 95% confidence interval) declined from 5.0 (4.5-5.6) to 1.9 (1.7-2.1), the interquintile rate ratio diminished from 2.7 (1.8-4.2) to 1.7 (1.1- 2.6), and the interquintile rate difference decreased from 4.6 (2.8- 6.4) to 1.1 (0.2-1.9). INTERPRETATION: The income-related disparity in rates of death from cervical cancer as measured by rate ratios and rate differences diminished markedly in urban Canada from 1971 to 1996. Among the numerous factors that may have contributed to the decline (including decline in fertility and improvement in diet), one important factor was probably the implementation of effective screening programs. PMID- 15136548 TI - Displacement of Canada's largest public illicit drug market in response to a police crackdown. AB - BACKGROUND: Law enforcement is often used in an effort to reduce the social, community and health-related harms of illicit drug use by injection drug users (IDUs). There are, however, few data on the benefits of such enforcement or on the potential harms. A large-scale police "crackdown" to control illicit drug use in Vancouver's Downtown Eastside provided us with an opportunity to evaluate the effect. METHODS: As part of our ongoing prospective cohort study of IDUs in Vancouver, we examined data collected from 244 IDUs in the 3 months before the police crackdown and from 142 IDUs in the 3 months after the start of the crackdown, on Apr. 7, 2003. All study subjects were active drug users. We also examined external data on needle exchanges and syringe disposal. RESULTS: The 2 groups of IDUs were statistically similar: they were mainly young (mean age 39 years) and male (63%), and they had injected illicit drugs for 13 years on average. Ethnic background and the proportion homeless were also similar. There were no statistically significant reported differences (all p > 0.1) in the street price of heroin, cocaine or "crack" in the 2 periods. In the 3-month periods before and after the crackdown, respectively, the rates of daily heroin injection were 27.9% and 26.8%, daily cocaine injection 28.7% and 27.5%, and daily crack use 59.4% and 60.6% (all p > 0.1). The proportions of study subjects receiving methadone treatment, 41.0% and 44.4% (p = 0.516), did not differ. However, the proportions reporting a change in where drugs were used, 22.5% and 33.8% (p < 0.05), and the proportions reporting a change in the neighbourhood of use because of police presence, 18.1% and 26.8% (p < 0.05), increased significantly. Needle-exchange data confirmed that the community levels of drug use were unchanged. Disposal statistics demonstrated that the monthly average number of used syringes found on the streets outside the traditional area of drug use increased from 784 in the 3 months before Apr. 1 to 1253 in the subsequent 3 months (p = 0.002) and the monthly average number of used syringes found in public boxes for the safe disposal of syringes decreased from 865 to 502 (p = 0.018). INTERPRETATION: The effort to control illicit drug use did not alter the price of drugs or the frequency of use, nor did it encourage enrollment in methadone treatment programs. Several measures indicated displacement of injection drug use from the area of the crackdown into adjacent areas of the city, which has implications for both recruitment of new initiates into injection drug use and HIV prevention efforts. PMID- 15136549 TI - Dieting among preadolescent and young adolescent females. AB - We examined the prevalence of dieting and negative eating attitudes among 2279 females (aged 10-14 years) in southern Ontario. Self-report questionnaires revealed that 29.3% of the girls were currently trying to lose weight and 10.5% had scores greater than the clinical threshold for disordered eating (Children's version of the Eating Attitudes Test [ChEAT] score > or = 20). Those with elevated ChEAT scores were more likely than those with lower scores to be engaged in dieting and other extreme weight control methods. PMID- 15136550 TI - C-reactive protein for the prediction of cardiovascular risk: ready for prime time? PMID- 15136551 TI - Generic isotretinoin: a new risk for unborn children. PMID- 15136552 TI - Hematopoietic stem cell transplantation: a primer for the primary care physician. AB - Hematopoietic stem cell transplantation has been used for many years to treat various malignant and nonmalignant hematologic conditions. However, the high-dose conditioning regimen can lead to major organ dysfunction, life-threatening infection and bleeding. In the allogeneic setting, graft-versus-host disease may also develop, making post-transplant management complex. Once a transplant recipient is discharged from hospital and returns to his or her local community, the primary care physician can play an important role in care. Recipients of stem cell transplants may be severely immunocompromised for many months after transplantation, especially if they are still taking immunosuppressive drugs. Furthermore, endocrine and metabolic deficiencies can develop, and transplant survivors are at risk of a second malignant disease. This review is intended as a basic overview of allogeneic and autologous stem cell transplantation with a special focus on long-term follow-up issues relevant to primary care providers. PMID- 15136555 TI - High and low affinity carbohydrate ligands revealed for murine SIGN-R1 by carbohydrate array and cell binding approaches, and differing specificities for SIGN-R3 and langerin. AB - The number of receptors of the 'C-type' lectin family is greater than previously thought with a considerable proportion on cells (dendritic cells and macrophages) critical for innate immunity. Establishing that they bind carbohydrates, unravelling and comparing details of their ligands is crucial for understanding the molecular basis of the cell-cell and cell-pathogen interactions that they mediate. Here we use carbohydrate arrays as a new approach to discovering the ligands of three recently described C-type lectin-type receptors on antigen presenting cells: murine SIGN-R1, SIGN-R3 and langerin. The arrays encompass an extensive panel including polysaccharides, glycoproteins, oligosaccharides and monosaccharides. These are probed with soluble forms of the receptors (IgG-Fc chimeras). The dominant specificities found for SIGN-R1 and SIGN-R3 are mannose- and fucose-related, as expressed on high mannose type N-glycans and Lewis(a/b)/Lewis(x/y)-type sequences, respectively, with subtle differences between the receptors. The dominant specificity for langerin is unique so far: a Lewisx-related sequence with sulfate at position 6 of the terminal galactose. The polysaccharide dextran, known from classical studies to elicit a T-independent response, and whose cellular uptake has been shown recently to be mediated by membrane-associated SIGN-R1, gave no binding signals with the soluble form of the protein. We highlight here the additional need for cell-based assays for detecting biologically relevant low affinity ligands, for we show with SIGN-R1 transfected cells that dextran is such a low affinity ligand for SIGN-R1 that binding is detectable only with the cell membrane-associated receptor. But there is a close relationship between dextran recognition and mannose/fucose recognition, with dextran- and mannose-conjugates co-localizing in intracellular compartments. PMID- 15136556 TI - Proteins of the Ikaros family control dendritic cell maturation required to induce optimal Th1 T cell differentiation. AB - Ikaros proteins are pleiotropic regulators of hematopoiesis and are critically required for the production of lymphocyte and dendritic cell (DC) lineages in mice. Here, we asked if Ikaros proteins could also play a role in the late stages of dendritic cell differentiation. Nuclear Ikaros proteins were up-regulated during the in vitro differentiation of human monocytes into mature DC, suggesting potential implications in this process. To address this question, a dominant negative mutant Ikaros isoform IK7 was over-expressed by retroviral gene transfer in human DC precursor cells, to interfere with the function of Ikaros family members during DC development. Expression of IK7 in CD34+ cells inhibited the production of IL-12-producing APCs. The resulting progeny of CD34+ cells and in particular, committed CD1a+ DC or CD14+ cell-derived DC, expressed low levels of MHC class II antigens and of the CD83 maturation marker on the cell surface. Such IK7-expressing DC induced naive allogeneic T cells to produce Th2 cytokines. Our results therefore delineate a new role for Ikaros family members, showing that normal levels of Ikaros proteins are essential in DC to regulate the terminal stages of maturation and the capacity to induce optimal Th1 T cell responses. PMID- 15136557 TI - MHC class II-independent and -dependent T cell expansion and B cell hyperactivity in vivo in mice deficient in CD152 (CTLA-4). AB - One of the key downregulators of T cell activation is CD152 (CTLA-4). Mice genetically deficient in CD152 (cd152(-/-) mice) develop massive expansion of both CD4(+) and CD8(+) T cells as well as increased numbers of splenic Ig secreting cells and serum Ig levels. To determine the dependence of the lymphoproliferation and B cell hyperactivity on MHC class II (MHCII), MHCII deficient (mhcii(-/-)) cd152(-/-) mice were generated. Compared to that in their mhcii(+/+) counterparts, expansion of CD4(+) cells in mhcii(-/-)cd152(-/-) mice was markedly attenuated. Nonetheless, expansion of CD8(+) cells was identical in both sets of mice, demonstrating that the effects of CD152 deficiency on CD4(+) cells can quantitatively be dissociated from those on CD8(+) cells, and pointing to a critical downregulatory role for CD152 in MHCII-independent CD8(+) cell activation in vivo. B cell hyperactivity also developed in mhcii(-/-)cd152(-/-) mice, albeit in a manner less rapid and less intense than that in their mhcii(+/+) counterparts, demonstrating an underlying MHCII-independent diathesis to B cell dysregulation and pointing to a critical downregulatory role for CD152 in MHCII-independent B cell activation in vivo. When human DQ8 was introduced as a transgene into mhcii(-/-)cd152(-/-) mice, B cell hyperactivity was restored to levels observed in mhcii(+/+)cd152(-/-) mice, pointing to a critical downregulatory role for CD152 in MHCII-dependent B cell activation in vivo superimposed upon its downregulatory role on MHCII-independent B cell activation. PMID- 15136558 TI - Chronic management of blood pressure after stroke. PMID- 15136559 TI - Complement 3 is involved in the synthetic phenotype and exaggerated growth of vascular smooth muscle cells from spontaneously hypertensive rats. AB - Vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) show the synthetic phenotype and exaggerated growth in comparison with VSMCs from normotensive Wistar-Kyoto (WKY) rats. We investigated genes associated with the synthetic phenotype and exaggerated growth of VSMCs from SHR by microarray. Expression of 1300 transcripts was evaluated by microarray with total mRNA extracted from mid-layer aortic smooth muscle of 3-week-old SHR/Izumo and WKY/Izumo rats. mRNAs encoding sodium-dependent neurotransmitter transporter, epidermal growth factor precursor, EEF2, leptin receptor long-isoform b, clathrin assembly protein short form, and preprocomplement 3 (pre-pro-C3) were expressed only in aortic smooth muscle from SHR by microarray and by reverse-transcription polymerase chain reaction analysis. Pre-pro-C3 mRNA was detected only in cultured VSMCs from SHR. Exogenous C3 changed VSMCs to the synthetic phenotype. Antisense oligodeoxynucleotides (ODN) to C3 reduced the higher level of DNA synthesis in VSMCs from SHR. Antisense ODN to C3 increased expression of SM22alpha mRNA and decreased expression of osteopontin and matrix Gla mRNAs. It also decreased expression of growth factor mRNAs in VSMCs from SHR. In conclusion, we have shown that C3, independent of other complement molecules, has direct effects on the phenotype of VSMCs and stimulates growth of these cells. C3 is produced only by VSMCs from SHR. Therefore, C3 may be the gene underlying the synthetic phenotype and exaggerated growth of VSMCs from SHR. C3 may be a new target for the treatment of hypertension. PMID- 15136561 TI - Role of protein phosphatase type 1 in contractile functions: myosin phosphatase. PMID- 15136562 TI - Hepatitis C virus glycoprotein E2 contains a membrane-proximal heptad repeat sequence that is essential for E1E2 glycoprotein heterodimerization and viral entry. AB - The E1 and E2 glycoproteins of hepatitis C virus form a noncovalently associated heterodimer that mediates viral entry. Glycoprotein E2 comprises a receptor binding domain (residues 384-661) that is connected to the transmembrane domain (residues 716-746) via a highly conserved sequence containing a hydrophobic heptad repeat (residues 675-699). Alanine- and proline-scanning mutagenesis of the E2 heptad repeat revealed that Leu675, Ser678, Leu689, and Leu692 are important for E1E2 heterodimerization. Furthermore, Pro and Ala substitution of all but one heptad repeat residue (Ser678) blocked the entry of E1E2-HIV-1 pseudotypes into Huh7 cells, irrespective of an effect on heterodimerization. Two conserved prolines (Pro676 and Pro683), occupying consecutive b positions of the heptad, were not required for E1E2 heterodimerization; however, Pro683 was critical for viral entry. Thus, disruption of the predicted alpha-helical structure by proline at position 683 is important for E2 function. The inability of mutants to mediate viral entry was not explained by a loss of receptor binding function, because all mutants were able to interact with a recombinant form of the CD81 large extracellular loop. Chimeras formed between the E1 and E2 ectodomains and the transmembrane domains of flavivirus prM and E glycoproteins, respectively, were able to heterodimerize, although with lower efficiency in comparison with wild type E1E2. The heptad repeat of E2 therefore requires the native transmembrane domain for full heterodimerization and viral entry function. Our data indicate that the membraneproximal heptad repeat of E2 is functionally homologous to the stem of flavivirus E glycoproteins. We propose that E2 has mechanistic features in common with class II fusion proteins. PMID- 15136563 TI - A signaling role of histone-binding proteins and INHAT subunits pp32 and Set/TAF Ibeta in integrating chromatin hypoacetylation and transcriptional repression. AB - Various post-translational modifications of histones significantly influence gene transcription. Although un- or hypoacetylated histones are tightly linked to transcriptional repression, the mechanisms and identities of chromatin signal transducer proteins integrating histone hypoacetylation into repression in humans have remained largely unknown. Here we show that the mammalian histone-binding proteins and inhibitor of acetyltransferases (INHAT) complex subunits, Set/template-activating factor-Ibeta (TAF-Ibeta) and pp32, specifically bind to unacetylated, hypoacetylated, and repressively marked histones but not to hyperacetylated histones. Additionally, Set/TAF-Ibeta and pp32 associate with histone deacetylases in vitro and in vivo and repress transcription from a chromatin-integrated template in vivo. Finally, Set/TAF-Ibeta and pp32 associate with an endogenous estrogen receptor-regulated gene, EB1, in the hypoacetylated transcriptionally inactive state but not with the hyperacetylated transcriptionally active form. Together, these data define a novel in vivo mechanistic role for the mammalian Set/TAF-Ibeta and pp32 proteins as transducers of chromatin signaling by integrating chromatin hypoacetylation and transcriptional repression. PMID- 15136564 TI - c-Fos phosphorylation induced by H2O2 prevents proteasomal degradation of c-Fos in cardiomyocytes. AB - Oxidants cause activation of the AP-1 transcription factor in cardiomyocytes. c Fos, a component of the AP-1 transcription factor, is transiently induced by H2O2 and the induction is sensitive to the protein synthesis inhibitor cycloheximide. With high percentage gel electrophoresis, multiple c-Fos bands were resolved by Western blot analyses, indicating post-translational modification of newly synthesized c-Fos protein after H2O2 exposure. Treatment of immunoprecipitated c Fos protein with the type 2 serine/threonine phosphatase A (PP2A) and immunoblotting of c-Fos protein with antibodies against phosphorylated serine or threonine demonstrated that c-Fos was phosphorylated at serine residues. A pharmacological inhibitor of JNKs inhibited the formation of multiple c-Fos bands without affecting c-fos transcription. The proteasomal inhibitor MG132 and Proteasome Inhibitor I extended the time course of c-Fos protein elevation. An increase in ubiquitin was detectable in c-Fos protein from H2O2-treated cells. Interestingly, treating the whole cell lysates with PP2A, but not calcineurin (i.e. PP2B), resulted in disappearance of c-Fos protein and MG132 was able to prevent this loss. H2O2 caused an elevation of PP2B and total phosphatase activity. The phosphatase inhibitor okadaic acid, but not PP2B inhibiter cypermethrin, extended the time course of c-Fos protein elevation after H2O2 exposure. These data suggest that JNK-mediated phosphorylation of newly synthesized c-Fos protects the protein from being degraded by the proteasome. PP2B independent dephosphorylation contributes to degradation of c-Fos protein during oxidative stress response of cardiomyocytes. PMID- 15136565 TI - CGI-58 interacts with perilipin and is localized to lipid droplets. Possible involvement of CGI-58 mislocalization in Chanarin-Dorfman syndrome. AB - Lipid droplets (LDs) are a class of ubiquitous cellular organelles that are involved in lipid storage and metabolism. Although the mechanisms of the biogenesis of LDs are still unclear, a set of proteins called the PAT domain family have been characterized as factors associating with LDs. Perilipin, a member of this family, is expressed exclusively in the adipose tissue and regulates the breakdown of triacylglycerol in LDs via its phosphorylation. In this study, we used a yeast two-hybrid system to examine the potential function of perilipin. We found direct interaction between perilipin and CGI-58, a deficiency of which correlated with the pathogenesis of Chanarin-Dorfman syndrome (CDS). Endogenous CGI-58 was distributed predominantly on the surface of LDs in differentiated 3T3-L1 cells, and its expression increased during adipocyte differentiation. Overexpressed CGI-58 tagged with GFP gathered at the surface of LDs and colocalized with perilipin. This interaction seems physiologically important because CGI-58 mutants carrying an amino acid substitution identical to that found in CDS lost the ability to be recruited to LDs. These mutations significantly weakened the binding of CGI-58 with perilipin, indicating that the loss of this interaction is involved in the etiology of CDS. Furthermore, we identified CGI-58 as a binding partner of ADRP, another PAT domain protein expressed ubiquitously, by yeast two-hybrid assay. GFP-CGI-58 expressed in non differentiated 3T3-L1 or CHO-K1 cells was colocalized with ADRP, and the CGI-58 mutants were not recruited to LDs carrying ADRP, indicating that CGI-58 may also cooperate with ADRP. PMID- 15136566 TI - Two pathways for store-mediated calcium entry differentially dependent on the actin cytoskeleton in human platelets. AB - A major pathway for stimulated Ca(2+) entry in non-excitable cells is activated following depletion of intracellular Ca(2+) stores. Secretion-like coupling between elements in the plasma membrane (PM) and Ca(2+) stores has been proposed as the most likely mechanism to activate this store-mediated Ca(2+) entry (SMCE) in several cell types. Here we identify two mechanisms for SMCE in human platelets activated by depletion of two independent Ca(2+) pools, which are differentially modulated by the actin cytoskeleton. Ca(2+) entry induced by depletion of a 2,5-di-(tert-butyl)-1,4-hydroquinone (TBHQ)-sensitive pool is increased by disassembly of the actin cytoskeleton and that induced by a TBHQ insensitive pool is reduced. Stabilization of the actin cytoskeleton prevented Ca(2+) entry by both mechanisms. We propose that the membrane-associated actin network prevents constitutive Ca(2+) entry via both pathways. Reorganization of the actin cytoskeleton permits the activation of Ca(2+) entry via both mechanisms, but only SMCE activated by the TBHQ-insensitive pool requires new actin polymerization, which may support membrane trafficking toward the PM. PMID- 15136567 TI - Disinactivation of N-type inactivation of voltage-gated K channels by an erbstatin analogue. AB - In some A-type voltage-gated K channels, rapid inactivation is achieved through the binding of an N-terminal domain of the pore-forming alpha-subunit or an associated beta-subunit to a cytoplasmic acceptor located at or near the channel pore using the ball-and-chain machinery (1-5). This inactivation involving the N terminus is known as N-type inactivation. Here, we describe an erbstatin (Erb) analogue as a small molecule inhibitor of the N-type inactivation in channels of Kv1.4 and Kv1.1+Kvbeta1. We show that this inhibition of inactivation (designated as "disinactivation") is potent and selective for N-type inactivation in heterologous cells (Chinese hamster ovary and Xenopus oocytes) expressing these A type channels. In Chinese hamster ovary cells, Erb increased the inactivation time constant of Kv1.4 from 86.5 +/- 9.5 to 150 +/- 10 ms (n = 6, p < 0.0 1). Similarly, Erb increased the inactivation time constant of Kv1.1+Kvbeta1 from 10 +/- 0.9 to 49.3 +/- 7 ms (n = 7, p < 0.01). The EC(50) for disinactivating Kv1.1+Kvbeta1 was 10.4 +/- 0.9 microm (n = 2-9). Erb had no effect upon another A channel, Kv4.3, which does not utilize the ball-and-chain mechanism. The mechanism of Erb-induced disinactivation was also investigated. Neither cysteine oxidation nor tyrosine kinase inhibition was involved. The results demonstrate that Erb can be used as a base structure to identify potent, selective small molecule inhibitors of intracellular protein-protein interactions, and that these disinactivators may offer another therapeutic approach to the treatment of seizure disorders. PMID- 15136568 TI - Probing the function of STAS domains of the Arabidopsis sulfate transporters. AB - Sulfate transporters in plants and animals are structurally conserved and have an amino-terminal domain that functions in transport and a carboxyl-terminal region that has been designated the STAS domain. The STAS domain in sulfate transporters has significant similarity to bacterial anti-sigma factor antagonists. To determine if the STAS domain has a role in controlling the activity of sulfate transporters, their stability, or their localization to the plasma membrane, we examined the effect of deleting or modifying the STAS domain of dominant sulfate transporters in roots of Arabidopsis thaliana. The A. thaliana Sultr1;2 and Sultr1;1 sulfate transporters rescue the methionine-dependent growth phenotype of the yeast sulfate transporter mutant strain CP154-7B. Constructs of Sultr1;2 in which the STAS domain was deleted (DeltaSTAS) resulted in synthesis of a truncated polypeptide that was unable to rescue the CP154-7B phenotype. The inability of these constructs to rescue the mutant phenotype probably reflected both low level cellular accumulation of the transporter and the inability of the truncated protein to localize to the plasma membrane. Fusing the STAS domain from other sulfate transporters to Sultr1;2 DeltaSTAS constructs restored elevated accumulation and plasma membrane localization, although the kinetics of sulfate uptake in the transformants were markedly altered with respect to transformants synthesizing wild-type Sultr1;2 protein. These results suggest that the STAS domain is essential, either directly or indirectly, for facilitating localization of the transporters to the plasma membrane, but it also appears to influence the kinetic properties of the catalytic domain of transporters. PMID- 15136569 TI - Structure and function of a hypothetical Pseudomonas aeruginosa protein PA1167 classified into family PL-7: a novel alginate lyase with a beta-sandwich fold. AB - Structural and functional analyses of alginate lyases are important in the clarification of the biofilm-dependent ecosystem in Pseudomonas aeruginosa and in the development of therapeutic agents for bacterial disease. Most alginate lyases are classified into polysaccharide lyase (PL) family-5 and -7 based on their primary structures. Family PL-7 enzymes are still poorly characterized especially in structural properties. Among family PL-7, a gene coding for a hypothetical protein (PA1167) homologous to Sphingomonas alginate lyase A1-II was found to be present in the P. aeruginosa genome. PA1167 overexpressed in Escherichia coli cleaved glycosidic bonds in alginate and released unsaturated saccharides, indicating that PA1167 is an alginate lyase catalyzing a beta-elimination reaction. The enzyme acted preferably on heteropolymeric regions endolytically and worked most efficiently at pH 8.5 and 40 degrees C. The specific activity of PA1167, however, was much weaker than that of the known alginate lyase AlgL, suggesting that AlgL plays a main role in alginate depolymerization in P. aeruginosa. In addition to this specific activity, differences were found between PA1167 and AlgL in enzyme properties such as molecular mass, optimum pH, salt effect, and substrate specificity. The first crystal structure of the family PL-7 alginate lyase was determined at 2.0 A resolution. PA1167 was found to form a glove-like beta-sandwich composed of 15 beta-strands and 3 alpha-helices. The structural difference between the beta-sandwich PA1167 of family PL-7 and alpha/alpha-barrel AlgL of family PL-5 may be responsible for the enzyme characteristics. Crystal structures of polysaccharide lyases determined so far indicate that they can be assigned to three folding groups having parallel beta helix, alpha/alpha-barrel, and alpha/alpha-barrel + antiparallel beta-sheet structures as basic frames. PA1167 is the fourth novel folding structure found among polysaccharide lyases. PMID- 15136570 TI - RNA editing generates tissue-specific sodium channels with distinct gating properties. AB - Sodium channels play an essential role in generating the action potential in eukaryotic cells, and their transcripts, especially those in insects, undergo extensive A-to-I RNA editing. The functional consequences of RNA editing of sodium channel transcripts, however, have yet to be determined. We characterized 20 splice variants of the German cockroach sodium channel gene BgNa(v). Functional analysis revealed that these variants exhibited a broad range of voltage-dependent activation and inactivation. Further analysis of two variants, BgNa(v)1-1 and BgNa(v)1-2, which activate at more depolarizing membrane potentials than other variants, showed that RNA editing events were responsible for variant-specific gating properties. Two U-to-C editing sites identified in BgNa(v)1-1 resulted in a Leu to Pro change in segment 1 of domain III (IIIS1) and a Val to Ala change in IVS4. The Leu to Pro change shifted both the voltage dependence of activation and steady-state inactivation in the depolarizing direction. Two A-to-I editing events in BgNa(v)1-2 resulted in a Lys to Arg change in IS2 and an Ile to Met change in IVS3. The Lys to Arg change shifted the voltage dependence of activation in the depolarizing direction. Moreover, these RNA editing events occurred in a tissue-specific and development-specific manner. Our findings provide direct evidence that RNA editing is an important mechanism generating tissue-/cell type-specific functional variants of sodium channels. PMID- 15136571 TI - Microtubule-associated protein light chain 2 is a stargazin-AMPA receptor complex interacting protein in vivo. AB - The ataxic mutant mouse stargazer is a null mutant for stargazin, a protein involved in the regulation of cell surface trafficking and synaptic targeting of AMPA receptors. The extreme C terminus of stargazin (sequence, -TTPV), confers high affinity for PDZ domain-containing proteins e.g. PSD-95. Interaction with PDZ proteins enables stargazin to fulfill its role as an AMPA receptor synaptic targeting molecule but is not essential for its ability to influence AMPA receptor trafficking to the neuronal cell surface. Using the yeast-two hybrid approach we screened for proteins that interact with the intracellular C-terminal tail of stargazin. Positive interactors included PDZ domain-containing proteins e.g. SAP97, SAP102, and PIST. Interestingly, light chain 2 of microtubule associated protein 1 (LC2), which does not contain a PDZ domain, was also a strong interactor. This was shown to be a direct interaction that occurred upstream of the -TTPV sequence of stargazin. Immunoprecipitations of Triton X-100 soluble cerebellar extracts revealed that LC2 is pulled down not only by anti stargazin antibodies but also anti-GluR2 antibodies suggesting that stargazin and AMPA receptor subunits associate with LC2. Immunopurified full-length, native stargazin was shown to co-associate not only with GluR2 in vivo but also with full-length, native LC2. Indeed, LC2 co-associates with stargazin when part of a tripartite complex comprising LC2-stargazin-GluR2. Since this complex was extracted using Triton X-100 and was devoid of PSD95, SAP97, and actin we postulate that LC2 is involved in trafficking of AMPA receptors in cerebellar neurons before they are anchored at the synapse. PMID- 15136572 TI - Targeting of endothelial nitric-oxide synthase to the cytoplasmic face of the Golgi complex or plasma membrane regulates Akt- versus calcium-dependent mechanisms for nitric oxide release. AB - The heterogeneous localization of endothelial nitricoxide synthase (eNOS) on the Golgi complex versus the plasma membrane has made it difficult to dissect the regulation of each pool of enzyme. Here, we generated fusion proteins that specifically target the plasma membrane or cytoplasmic aspects of the Golgi complex and have assessed eNOS activation. Plasma membrane-targeted eNOS constructs were constitutively active, phosphorylated, and responsive to transmembrane calcium fluxes, yet were insensitive to further activation by Akt mediated phosphorylation. In contrast, cis-Golgi complex-targeted eNOS behaved similarly to wild-type eNOS and was less sensitive to calcium-dependent activation and highly responsive to Akt-dependent phosphorylation compared with plasma membrane versions. In plasma membrane- and Golgi complex-targeted constructs, Ser1179 is critical for NO production. This study provides clear evidence for functional roles of plasma membrane- and Golgi complex-localized eNOS and supports the concept that proteins thought to be regulated and to function exclusively in the plasma membrane of cells can indeed signal and be regulated in internal Golgi membranes. PMID- 15136573 TI - 5-oxo-6,8,11,14-eicosatetraenoic acid stimulates the release of the eosinophil survival factor granulocyte/macrophage colony-stimulating factor from monocytes. AB - Allergic diseases such as asthma are characterized by tissue eosinophilia induced by the combined effects of chemoattractants and cytokines. Lipid mediators are a major class of endogenous chemoattractants, among which 5-oxo-6,8,11,14 eicosatetraenoic acid (5-oxo-ETE) is the most potent for human eosinophils. In this study, we investigated the effects of 5-oxo-ETE on eosinophil survival by flow cytometry. We found that this compound could promote eosinophil survival in the presence of small numbers of contaminating monocytes, but not in their absence. The conditioned medium from monocytes treated for 24 h with 5-oxo-ETE also strongly promoted eosinophil survival, whereas the medium from vehicle treated monocytes had no effect. An antibody against the granulocyte/macrophage colony-stimulating factor (GM-CSF) completely blocked the response of eosinophils to the conditioned medium from 5-oxo-ETE-treated monocytes, whereas an antibody against interleukin-5 had no effect. Furthermore, 5-oxo-ETE stimulated the release of GM-CSF from cultured monocytes in amounts compatible with eosinophil survival activity, with a maximal effect being observed after 24 h. This effect was concentration-dependent and could be observed at concentrations in the picomolar range. 5-Oxo-ETE and leukotriene B(4) had similar effects on GM-CSF release at low concentrations, but 5-oxo-ETE induced a much stronger response at concentrations of 10 nm or higher. This is the first report that 5-oxo-ETE can induce the release of any cytokine, suggesting that it could be an important mediator in allergic and other inflammatory diseases due both to its chemoattractant properties and to its potent effects on the synthesis of the survival factor GM-CSF. PMID- 15136574 TI - Concerted action of diacetylchitobiose deacetylase and exo-beta-D-glucosaminidase in a novel chitinolytic pathway in the hyperthermophilic archaeon Thermococcus kodakaraensis KOD1. AB - The hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 possesses chitinase (Tk-ChiA) and exo-beta-D-glucosaminidase (Tk-GlmA) for chitin degradation; the former produces diacetylchitobiose (GlcNAc2) from chitin, and the latter hydrolyzes chitobiose (GlcN2) to glucosamine (GlcN). To identify the enzyme that physiologically links these two activities, here we focused on the deacetylase that provides the substrate for Tk-GlmA from GlcNAc2. The deacetylase could be detected in and partially purified from T. kodakaraensis cells, and the corresponding gene (Tk-dac) was identified on the genome. The deduced amino acid sequence was classified into the LmbE protein family including N acetylglucosaminylphosphatidylinositol de-N-acetylases and 1-D-myo-inosityl-2 acetamido-2-deoxy-alpha-D-glucopyranoside deacetylase. Recombinant Tk-Dac showed deacetylase activity toward N-acetylchitooligosaccharides (GlcNAc(2-5)), and the deacetylation site was revealed to be specific at the nonreducing GlcNAc residue. The enzyme also deacetylated GlcNAc monomer. In T. kodakaraensis cells, the transcription of Tk-dac, Tk-glmA, Tk-chiA, and the clustered genes were induced by GlcNAc2, suggesting the function of this gene cluster in chitin catabolism in vivo. These results have revealed a unique chitin catabolic pathway in T. kodakaraensis, in which GlcNAc2 produced from chitin is degraded by the concerted action of Tk-Dac and Tk-GlmA. That is, GlcNAc2 is site-specifically deacetylated to GlcN-GlcNAc by Tk-Dac and then hydrolyzed to GlcN and GlcNAc by Tk-GlmA followed by a second deacetylation step of the remaining GlcNAc by Tk-Dac to form GlcN. This is the first elucidation of an archaeal chitin catabolic pathway and defines a novel mechanism for dimer processing using a combination of deacetylation and cleavage, distinct from any previously known pathway. PMID- 15136575 TI - Localization of human Mcm10 is spatially and temporally regulated during the S phase. AB - Mcm10 (Dna43) is an essential protein for the initiation of DNA replication in Saccharomyces cerevisiae. Recently, we identified a human Mcm10 homolog and found that it is regulated by proteolysis and phosphorylation in a cell cycle-dependent manner and that it binds chromatin exclusively during the S phase of the cell cycle. However, the precise roles that Mcm10 plays are still unknown. To study the localization dynamics of human Mcm10, we established HeLa cell lines expressing green fluorescent protein (GFP)-tagged Mcm10. From early to mid-S phase, GFP-Mcm10 appeared in discrete nuclear foci. In early S phase, several hundred foci appeared throughout the nucleus. In mid-S phase, the foci appeared at the nuclear periphery and nucleolar regions. In the late S and G phases, GFP Mcm10 was localized to nucleoli. Although (2)the distributions of GFP-Mcm10 during the S phase resembled those of replication foci, GFP-Mcm10 foci did not colocalize with sites of DNA synthesis in most cases. Furthermore, the transition of GFP-Mcm10 distribution patterns preceded changes in replication foci patterns or proliferating cell nuclear antigen foci patterns by 30-60 min. These results suggest that human Mcm10 is temporarily recruited to the replication sites 30-60 min before they replicate and that it dissociates from chromatin after the activation of the prereplication complex. PMID- 15136576 TI - Specific residues of the GDP/GTP exchange factor Bud5p are involved in establishment of the cell type-specific budding pattern in yeast. AB - Cells of the budding yeast undergo oriented cell division by choosing a specific site for growth depending on their cell type. Haploid a and alpha cells bud in an axial pattern whereas diploid a/alpha cells bud in a bipolar pattern. The Ras like GTPase Rsr1p/Bud1p, its GDP-GTP exchange factor Bud5p, and its GTPase activating protein Bud2p are essential for selecting the proper site for polarized growth in all cell types. Here we showed that specific residues at the N terminus and the C terminus of Bud5p were important for bipolar budding, while some residues were involved in both axial and bipolar budding. These bipolar specific mutations of BUD5 disrupted proper localization of Bud5p in diploid a/alpha cells without affecting Bud5p localization in haploid alpha cells. In contrast, Bud5p expressed in the bud5 mutants defective in both budding patterns failed to localize in all cell types. Thus, these results identify specific residues of Bud5p that are likely to be involved in direct interaction with spatial landmarks, which recruit Bud5p to the proper bud site. Finally, we found a new start codon of BUD5, which extends the open reading frame to 210 bp upstream of the previously estimated start site, thus encoding a polypeptide of 608 amino acid residues. Bud5p with these additional N-terminal residues interacted with Bud8p, a potential bipolar landmark, suggesting that the N terminal region is necessary for recognition of the spatial cues. PMID- 15136577 TI - Two conserved cysteine triads in human Ero1alpha cooperate for efficient disulfide bond formation in the endoplasmic reticulum. AB - Human Ero1alpha is an endoplasmic reticulum (ER)-resident protein responsible for protein disulfide isomerase (PDI) oxidation. To clarify the molecular mechanisms underlying its function, we generated a panel of cysteine replacement mutants and analyzed their capability of: 1) complementing a temperature-sensitive yeast Ero1 mutant, 2) favoring oxidative folding in mammalian cells, 3) forming mixed disulfides with PDI and ERp44, and 4) adopting characteristic redox-dependent conformations. Our results reveal that two essential cysteine triads (Cys85-Cys94 Cys99 and Cys391-Cys394-Cys397) cooperate in electron transfer, with Cys94 likely forming mixed disulfides with PDI. Dominant negative phenotypes arise when critical residues within the triads are mutated (Cys394, Cys397, and to a lesser extent Cys99). Replacing the first cysteine in either triad (Cys85 or Cys391) generates mutants with weaker activity. In addition, mutating either Cys85 or Cys391, but not Cys397, reverts the dominant negative phenotype of the C394A mutant. These findings suggest that interactions between the two triads, dependent on Cys85 and Cys391, are important for Ero1alpha function, possibly stabilizing a platform for efficient PDI oxidation. PMID- 15136578 TI - Biosynthetic processing of the Pro-alpha1(V)Pro-alpha2(V)Pro-alpha3(V) procollagen heterotrimer. AB - Type V collagen is a quantitatively minor fibrillar collagen comprised of different chain compositions in different tissues. The most widely distributed form, an alpha1(V)2alpha2(V) heterotrimer, regulates the physical properties of type I/V heterotypic collagen fibrils via partially processed NH2-terminal globular sequences. A less characterized alpha1(V)alpha2(V)alpha3(V) heterotrimer has a much more limited distribution of expression and unknown function(s). We characterized the biosynthetic processing of pro-alpha1(V)2pro-alpha2(V) procollagen previously and showed it to differ in important ways from biosynthetic processing of the major fibrillar procollagens I-III. Here we have successfully produced recombinant pro-alpha1(V)pro-alpha2(V)pro-alpha3(V) heterotrimers. We use these, and mouse embryo fibroblasts doubly homozygous null for the Bmp1 gene, which encodes the metalloproteinase bone morphogenetic protein 1 (BMP-1), and for a gene encoding the closely related metalloproteinase mammalian Tolloid-like 1, to characterize biosynthetic processing of pro alpha1(V)pro-alpha2(V)pro-alpha3(V) heterotrimers, thus completing characterization of type V collagen biosynthetic processing. Whereas pro alpha1(V) and pro-alpha2(V) processing in pro-alpha1(V)pro-alpha2(V)pro-alpha3(V) heterotrimers is similar to that which occurs in pro-alpha1(V)2pro-alpha2(V) heterotrimers, the processing of pro-alpha3(V) by BMP-1 occurs at an unexpected site within NH2-terminal globular sequences. We also demonstrate that, despite similarities in NH2-terminal domain structures, pro-alpha2(V) NH2-terminal globular sequences are not cleaved by ADAMTS-2, the metalloproteinase that cleaves the N-propeptides of the major fibrillar procollagen chains. PMID- 15136579 TI - Visualization of G protein betagamma dimers using bimolecular fluorescence complementation demonstrates roles for both beta and gamma in subcellular targeting. AB - To investigate the role of subcellular localization in regulating the specificity of G protein betagamma signaling, we have applied the strategy of bimolecular fluorescence complementation (BiFC) to visualize betagamma dimers in vivo. We fused an amino-terminal yellow fluorescent protein fragment to beta and a carboxyl-terminal yellow fluorescent protein fragment to gamma. When expressed together, these two proteins produced a fluorescent signal in human embryonic kidney 293 cells that was not obtained with either subunit alone. Fluorescence was dependent on betagamma assembly in that it was not obtained using beta2 and gamma1, which do not form a functional dimer. In addition to assembly, BiFC betagamma complexes were functional as demonstrated by more specific plasma membrane labeling than was obtained with individually tagged fluorescent beta and gamma subunits and by their abilities to potentiate activation of adenylyl cyclase by alpha(s) in COS-7 cells. To investigate isoform-dependent targeting specificity, the localization patterns of dimers formed by pair-wise combinations of three different beta subunits with three different gamma subunits were compared. BiFC betagamma complexes containing either beta1 or beta2 localized to the plasma membrane, whereas those containing beta5 accumulated in the cytosol or on intracellular membranes. These results indicate that the beta subunit can direct trafficking of the gamma subunit. Taken together with previous observations, these results show that the G protein alpha, beta, and gamma subunits all play roles in targeting each other. This method of specifically visualizing betagamma dimers will have many applications in sorting out roles for particular betagamma complexes in a wide variety of cell types. PMID- 15136580 TI - Exploring the collagen-binding site of the DDR1 tyrosine kinase receptor. AB - Discoidin domain receptors 1 and 2 (DDR1 and DDR2) are tyrosine kinase receptors activated by triple-helical collagens. Aberrant expression and signaling of these receptors have been implicated in several human diseases linked to accelerated matrix degradation and remodeling including tumor invasion, atherosclerosis and liver fibrosis. The objective of this study is to characterize the collagen binding sites in the discoidin domains of DDR1 and DDR2 at a molecular level. We expressed glutathione S-transferase fusion proteins containing the discoidin and extracellular domains of DDR1 and DDR2 in insect cells and subjected them to a solid-phase collagen-binding assay. We found high affinity binding of the DDR extracellular domains to immobilized type I collagen and confirmed the discoidin collagen interaction with an enzyme-linked immunosorbent assay-based read-out. Furthermore, we created a three-dimensional model of the DDR1 discoidin domain based on the related domains of blood coagulation factors V and VIII. This model predicts the presence of four neighboring, surface-exposed loops that are topologically equivalent to a major phospholipid-binding site in factors V and VIII. To test the involvement of these loops in collagen binding, we mutated individual amino acid residues to alanine or deleted short sequence stretches within these loops. We found that several residues within loop 1 (Ser-52-Thr-57) and loop 3 (Arg-105-Lys-112) as well as Ser-175 in loop 4 are critically involved in collagen binding. Our structure-function analysis of the DDR discoidin domains provides new insights into this non-integrin-mediated collagen-signaling mechanism and may ultimately lead to the design of small molecule inhibitors that interfere with aberrant DDR function. PMID- 15136581 TI - Identification of strain-specific variants of mouse Adamts13 gene encoding von Willebrand factor-cleaving protease. AB - Human ADAMTS13 was recently identified as a gene encoding von Willebrand factor cleaving protease, hADAMTS13. Both congenital and acquired defects in this enzyme can cause thrombotic thrombocytopenic purpura. hADAMTS13 consists of 1,427 amino acid residues and is composed of multiple structural domains including thrombospondin type 1 motifs and CUB domains. To analyze the functional roles of these domains in vivo, we determined the cDNA sequence of the mouse ortholog, mADAMTS13. Unexpectedly, two forms of the mouse Adamts13 gene were isolated that differed in the insertion of an intracisternal A particle (IAP) retrotransposon including a premature stop codon. The IAP insertion was found in BALB/c, C3H/He, C57BL/6, and DBA/2 strains but not in the 129/Sv strain. The outbred ICR strain had either the IAP-free or IAP-inserted allele or both. IAP-free Adamts13 encoded mADAMTS13L, a protein of 1,426 amino acid residues with the same domain organization as hADAMTS13. In contrast, IAP-inserted Adamts13 encoded a C terminally truncated enzyme, mADAMTS13S, that is comprised of only 1,037 amino acid residues and lacking the C-terminal two thrombospondin type 1 motifs and two CUB domains. Strain specificity was also confirmed by reverse transcription-PCR and Northern blot analyses. Both recombinant mADAMTS13L and mADAMTS13S exhibited von Willebrand factor cleaving activities in vitro. The natural variation in mouse ADAMTS13 should allow for the determination of hitherto unknown functions of its C-terminal domains in vivo. PMID- 15136582 TI - The structure of the pantothenate kinase.ADP.pantothenate ternary complex reveals the relationship between the binding sites for substrate, allosteric regulator, and antimetabolites. AB - Pantothenate kinase catalyzes the first step in the biosynthesis of coenzyme A, the major acyl group carrier in biology. In bacteria, regulation of pantothenate kinase activity is a major factor in controlling intracellular coenzyme A levels, and pantothenate analogs are growth-inhibiting antimetabolites. We have extended the structural information on Escherichia coli pantothenate kinase by determining the structure of the enzyme.ADP. pantothenate ternary complex. Pantothenate binding induces a significant conformational change in amino acids 243-263, which form a "lid" that folds over the open pantothenate binding groove. The positioning of the substrates suggests the reaction proceeds by a concerted mechanism that involves a dissociative transition state, although the negative charge neutralization of the gamma-phosphate by Arg-243, Lys-101, and Mg(2+) coupled with hydrogen bonding of the C1 of pantothenate to Asp-127 suggests different interpretations of the phosphoryl transfer mechanism of pantothenate kinase. N-alkylpantothenamides are substrates for pantothenate kinase. Modeling these antimetabolites into the pantothenate active site predicts that they bind in the same orientation as pantothenate with their alkyl chains interacting with the hydrophobic dome over the pantothenate pocket, which is also accessed by the beta-mercaptoethylamine moiety of the allosteric regulator, coenzyme A. These structural/biochemical studies illustrate the intimate relationship between the substrate, allosteric regulator, and antimetabolite binding sites on pantothenate kinase and provide a framework for studies of its catalysis and feedback regulation. PMID- 15136583 TI - Crystallographic and biophysical analysis of a bacterial signal peptidase in complex with a lipopeptide-based inhibitor. AB - We report here the crystallographic and biophysical analysis of a soluble, catalytically active fragment of the Escherichia coli type I signal peptidase (SPase Delta2-75) in complex with arylomycin A2. The 2.5-A resolution structure revealed that the inhibitor is positioned with its COOH-terminal carboxylate oxygen (O45) within hydrogen bonding distance of all the functional groups in the catalytic center of the enzyme (Ser90 O-gamma, Lys145 N-zeta, and Ser88 O-gamma) and that it makes beta-sheet type interactions with the beta-strands that line each side of the binding site. Ligand binding studies, calorimetry, fluorescence spectroscopy, and stopped-flow kinetics were also used to analyze the binding mode of this unique non-covalently bound inhibitor. The crystal structure was solved in the space group P4(3)2(1)2. A detailed comparison is made to the previously published acyl-enzyme inhibitor complex structure (space group: P2(1)2(1)2) and the apo-enzyme structure (space group: P4(1)2(1)2). Together this work provides insights into the binding of pre-protein substrates to signal peptidase and will prove helpful in the development of novel antibiotics. PMID- 15136584 TI - Trypanosoma congolense infections: antibody-mediated phagocytosis by Kupffer cells. AB - Immunohistochemical double-label technique was used to detect trypanosomal antigen in macrophages. Immunoglobulin (Ig)M as well as IgG2a monoclonal antibodies (mAb) specific for the variant surface glycoprotein (VSG) mediated phagocytosis of Trypanosoma congolense variant antigenic type (VAT) TC13 by macrophages [bone marrow-derived macrophage cell line from BALB/c (BALB.BM)] in vitro. Administration of these IgM or IgG2a antibodies to BALB/c mice 30 min after injection of 3 x 10(8) T. congolense mediated phagocytosis of trypanosomes by Kupffer cells of the liver within 1 h. Plasma levels of the monokines interleukin (IL)-1beta, IL-10, and IL-12p40 were significantly increased 6-48 h after phagocytosis. In BALB/c mice infected with 10(3) T. congolense, a small degree of phagocytosis of trypanosomes by Kupffer cells, mediated by actively synthesized antibodies, was detected as early as 5 days after infection. Phagocytosis of trypanosomes was dramatically enhanced on day 6. Concomitantly, the Kupffer cells trippled in size. In BALB/c mice infected for 6 days, treatment with IgM or IgG2a mAb specific for T. congolense VSG led to clearance of VAT TC13 parasitemia but did not prevent death at the second parasitemia of a different VAT. We conclude that IgM as well as IgG antibody mediate phagocytosis of trypanosomes by Kupffer cells. PMID- 15136585 TI - Localization of serglycin in human neutrophil granulocytes and their precursors. AB - Serglycin is a major proteoglycan of hematopoietic cells. It is thought to play a role in the packaging of granule proteins in human neutrophil granulocytes. The presence of serglycin in myeloid cells has been demonstrated only at the transcriptional level. We generated a polyclonal antibody against recombinant human serglycin. Here, we show the localization of serglycin in humans during neutrophil differentiation. Immunocytochemistry revealed serglycin immunoreactivity in the Golgi area of promyelocytes (PM) and myelocytes (MC), as well as in a few band cells and mature neutrophil granulocytes. Granular staining was detected near the Golgi apparatus in some of the PM, and the major part of the cytoplasm was negative. Immunoelectron microscopy showed serglycin immunoreactivity located to the Golgi apparatus and a few immature granules of PM and MC. The decreasing level of serglycin protein during myeloid differentiation coincided with a decrease of mRNA expression, as evaluated by Northern blotting. Subcellular fractions of neutrophil granulocytes were obtained. Serglycin immunoreactivity was detected in the fraction containing Golgi apparatus, plasma membrane, and secretory vesicles by Western blotting and enzyme-linked immunosorbent assay. Serglycin was not detected in subcellular fractions containing primary, secondary, or tertiary granules. Together, these findings indicate that serglycin is located to the Golgi apparatus and a few immature granules during neutrophil differentiation. This is consistent with a function for serglycin in formation of granules in neutrophil granulocytes. Our findings contrast the view that native serglycin is present in mature granules and plays a role in packaging and regulating the activity of proteolytic enzymes there. PMID- 15136586 TI - The monocyte Fcgamma receptors FcgammaRI/gamma and FcgammaRIIA differ in their interaction with Syk and with Src-related tyrosine kinases. AB - There are important differences in signaling between the Fc receptor for immunoglobulin G (IgG) FcgammaRIIA, which uses the Ig tyrosine-activating motif (ITAM) within its own cytoplasmic domain, and FcgammaRI, which transmits signals by means of an ITAM located within the cytoplasmic domain of its associated gamma chain. For example, in transfected epithelial cells and COS-1 cells, FcgammaRIIA mediates phagocytosis of IgG-coated red blood cells more efficiently than does FcgammaRI/gamma, and enhancement of phagocytosis by Syk kinase is more pronounced for FcgammaRI/gamma than for FcgammaRIIA. In addition, structure/function studies indicate that the gamma-chain ITAM and the FcgammaRIIA ITAM have different requirements for mediating the phagocytic signal. To study the differences between FcgammaRIIA and FcgammaRI/gamma, we examined the interaction of FcgammaRIIA and the FcgammaRI/gamma chimera FcgammaRI-gamma-gamma (extracellular domain-transmembrane domain-cytoplasmic domain) with Syk kinase and with the Src related tyrosine kinases (SRTKs) Hck and Lyn in transfected COS-1 cells. Our data indicate that FcgammaRIIA interacts more readily with Syk than does FcgammaRI gamma-gamma and suggest that one consequence may be the greater phagocytic efficiency of FcgammaRIIA compared with FcgammaRI/gamma. Furthermore, individual SRTKs affect the efficiency of phagocytosis differently for FcgammaRI-gamma-gamma and FcgammaRIIA and also influence the ability of these receptors to interact with Syk kinase. Taken together, the data suggest that differences in signaling by FcgammaRIIA and FcgammaRI-gamma-gamma are related in part to interaction with Syk and Src kinases and that individual SRTKs play different roles in FcgammaR mediated phagocytosis. PMID- 15136587 TI - TGF-beta regulation of human macrophage scavenger receptor CD163 is Smad3 dependent. AB - Tight regulation of the inflammatory response is essential for the maintenance of physiologic homeostasis. A potentially important mediator of this process is CD163, a macrophage-specific member of the scavenger receptor cysteine-rich family. CD163 surface expression is up-regulated by glucocorticoids and the anti inflammatory cytokine interleukin-10, and CD163 is shed acutely from the cell surface in response to lipopolysaccharide. We now demonstrate that transforming growth factor-beta (TGF-beta) markedly reduces expression of CD163. Treatment of primary human monocytes with TGF-beta inhibited basal as well as dexamethasone induced CD163 mRNA and protein expression. De novo protein synthesis was not required for this inhibition, suggesting that TGF-beta regulates CD163 expression transcriptionally. To delineate this transcriptional regulation, a 2.5-kb fragment of the CD163 promoter was isolated. This promoter was inhibited by TGF beta, and suppression was dependent on Smad3 expression. These results define a novel function for TGF-beta and implicate an important role for CD163 in the host response to inflammation. PMID- 15136588 TI - Uptake of Abeta 1-40- and Abeta 1-42-coated yeast by microglial cells: a role for LRP. AB - Artificial diffuse and amyloid core of neuritic plaques [beta-amyloid peptide (Abeta) deposits] could be prepared using heat-killed yeast particles opsonized with Abeta 1-40 or Abeta 1-42 peptides. Interaction and fate of these artificial deposits with microglial cells could be followed using a method of staining that allows discrimination of adherent and internalized, heat-killed yeast particles. Using this system, it was possible to show that nonfibrillar or fibrillar (f)Abeta peptides, formed in solution upon heating (aggregates), could not impair the internalization of heat-killed yeast particles opsonized with fAbeta 1-40 or fAbeta 1-42. This indicated that depending on their physical state, Abeta peptide(s) do not recognize the same receptors and probably do not follow the same internalization pathway. Using competitive ligands of class A scavenger receptors (SR-A) or low-density lipoprotein-related receptor protein (LRP), it has been shown that SR-A were not involved in the recognition of amyloid peptide deposits, whereas LRP specifically recognized deposits of fAbeta 1-42 (but not fAbeta 1-40) and mediated their phagocytosis. PMID- 15136589 TI - DNAM-1 and PVR regulate monocyte migration through endothelial junctions. AB - DNAX accessory molecule 1 (DNAM-1; CD226) is a transmembrane glycoprotein involved in T cell and natural killer (NK) cell cytotoxicity. We demonstrated recently that DNAM-1 triggers NK cell-mediated killing of tumor cells upon engagement by its two ligands, poliovirus receptor (PVR; CD155) and Nectin-2 (CD112). In the present paper, we show that PVR and Nectin-2 are expressed at cell junctions on primary vascular endothelial cells. Moreover, the specific binding of a soluble DNAM-1-Fc molecule was detected at endothelial junctions. This binding was almost completely abrogated by anti-PVR monoclonal antibodies (mAbs), but not modified by anti-Nectin-2 mAbs, which demonstrates that PVR is the major DNAM-1 ligand on endothelial cells. Because DNAM-1 is highly expressed on leukocytes, we investigated the role of the DNAM-1-PVR interaction during the monocyte transendothelial migration process. In vitro, both anti-DNAM-1 and anti PVR mAbs strongly blocked the transmigration of monocytes through the endothelium. Moreover, after anti-DNAM-1 or anti-PVR mAb treatment, monocytes were arrested at the apical surface of the endothelium over intercellular junctions, which strongly suggests that the DNAM-1-PVR interaction occurs during the diapedesis step. Altogether, our results demonstrate that DNAM-1 regulates monocyte extravasation via its interaction with PVR expressed at endothelial junctions on normal cells. PMID- 15136590 TI - Increased expression of human T lymphocyte virus type I (HTLV-I) Tax11-19 peptide human histocompatibility leukocyte antigen A*201 complexes on CD4+ CD25+ T Cells detected by peptide-specific, major histocompatibility complex-restricted antibodies in patients with HTLV-I-associated neurologic disease. AB - Human T lymphocyte virus type I (HTLV-I)-associated chronic inflammatory neurological disease (HTLV-I-associated myelopathy/tropical spastic paraparesis [HAM/TSP]) is suggested to be an immunopathologically mediated disorder characterized by large numbers of HTLV-I Tax-specific CD8+ T cells. The frequency of these cells in the peripheral blood and cerebrospinal fluid is proportional to the amount of HTLV-I proviral load and the levels of HTLV-I tax mRNA expression. As the stimulus for these virus-specific T cells are immunodominant peptide-human histocompatibility leukocyte antigen (HLA) complexes expressed on antigen presenting cells, it was of interest to determine which cells express these complexes and at what frequency. However, until now, it has not been possible to identify and/or quantify these peptide-HLA complexes. Using a recently developed antibody that specifically recognizes Tax11-19 peptide-HLA-A*201 complexes, the level of Tax11-19-HLA-A*201 expression on T cells was demonstrated to be increased in HAM/TSP and correlated with HTLV-I proviral DNA load, HTLV-I tax mRNA load, and HTLV-I Tax-specific CD8+ T cell frequencies. Furthermore, CD4+ CD25+ T cells were demonstrated to be the major reservoir of HTLV-I provirus as well as Tax11-19 peptide-HLA-A*201 complexes. These results indicate that the increased detection and visualization of peptide-HLA complexes in HAM/TSP CD4+ CD25+ T cell subsets that are shown to stimulate and expand HTLV-I Tax-specific CD8+ T cells may play an important role in the pathogenesis of HTLV-I-associated neurological disease. PMID- 15136591 TI - KLF2 Is a novel transcriptional regulator of endothelial proinflammatory activation. AB - The vascular endothelium is a critical regulator of vascular function. Diverse stimuli such as proinflammatory cytokines and hemodynamic forces modulate endothelial phenotype and thereby impact on the development of vascular disease states. Therefore, identification of the regulatory factors that mediate the effects of these stimuli on endothelial function is of considerable interest. Transcriptional profiling studies identified the Kruppel-like factor (KLF)2 as being inhibited by the inflammatory cytokine interleukin-1beta and induced by laminar shear stress in cultured human umbilical vein endothelial cells. Overexpression of KLF2 in umbilical vein endothelial cells robustly induced endothelial nitric oxide synthase expression and total enzymatic activity. In addition, KLF2 overexpression potently inhibited the induction of vascular cell adhesion molecule-1 and endothelial adhesion molecule E-selectin in response to various proinflammatory cytokines. Consistent with these observations, in vitro flow assays demonstrate that T cell attachment and rolling are markedly attenuated in endothelial monolayers transduced with KLF2. Finally, our studies implicate recruitment by KLF2 of the transcriptional coactivator cyclic AMP response element-binding protein (CBP/p300) as a unifying mechanism for these various effects. These data implicate KLF2 as a novel regulator of endothelial activation in response to proinflammatory stimuli. PMID- 15136592 TI - Surface sialic acids taken from the host allow trypanosome survival in tsetse fly vectors. AB - The African trypanosome Trypanosoma brucei, which causes sleeping sickness in humans and Nagana disease in livestock, is spread via blood-sucking Tsetse flies. In the fly's intestine, the trypanosomes survive digestive and trypanocidal environments, proliferate, and translocate into the salivary gland, where they become infectious to the next mammalian host. Here, we show that for successful survival in Tsetse flies, the trypanosomes use trans-sialidase to transfer sialic acids that they cannot synthesize from host's glycoconjugates to the glycosylphosphatidylinositols (GPIs), which are abundantly expressed on their surface. Trypanosomes lacking sialic acids due to a defective generation of GPI anchored trans-sialidase could not survive in the intestine, but regained the ability to survive when sialylated by means of soluble trans-sialidase. Thus, surface sialic acids appear to protect the parasites from the digestive and trypanocidal environments in the midgut of Tsetse flies. PMID- 15136593 TI - New target, new drug, old paradigm. PMID- 15136594 TI - Trawling for genes that predict response to breast cancer adjuvant therapy. PMID- 15136595 TI - Gene expression profiles predict complete pathologic response to neoadjuvant paclitaxel and fluorouracil, doxorubicin, and cyclophosphamide chemotherapy in breast cancer. AB - PURPOSE: The goal of this study was to examine the feasibility of developing a multigene predictor of pathologic complete response (pCR) to sequential weekly paclitaxel and fluorouracil + doxorubicin + cyclophosphamide (T/FAC) neoadjuvant chemotherapy regimen for breast cancer. PATIENTS AND METHODS: All patients underwent one-time pretreatment fine-needle aspiration to obtain RNA from the cancer for transcriptional profiling using cDNA arrays containing 30721 human sequence clones. Analysis was performed after profiling, and 42 patients' clinical results were available, 24 of which were used for predictive marker discovery; 18 patients' results were used as an independent validation set. RESULTS: Thirty-one percent of patients had pCR (six discovery and seven validation), defined as disappearance of all invasive cancer in the breast after completion of chemotherapy. We could identify no single marker that was sufficiently associated with pCR to be used as an individual predictor. A multigene model with 74 markers (P or= 0.5 (391 patients). The primary endpoint of the study was mortality at 6 mo after the index event. RESULTS: Six-month mortality was 5.1% (19 deaths) in the group with salvage index < 0.5, compared with 1.0% (4 deaths) in the group with salvage index >or= 0.5 (odds ratio, 5.1; 95% confidence interval, 1.9-13.3; P = 0.001). Salvage index (median [25th, 75th percentiles] was significantly smaller in nonsurvivors than in survivors (0.19 [0.05, 0.37] vs. 0.50 [0.26, 0.80], P = 0.0004). The Cox proportional hazards model showed that myocardial salvage index (P = 0.0007), initial perfusion defect (P = 0.0007), and age (P = 0.04) were independently associated with 6-mo mortality. CONCLUSION: Myocardial salvage achieved by reperfusion therapy predicts mortality in patients with acute myocardial infarction. Our findings support the use of salvage index as a surrogate of mortality in clinical trials designed to test the efficacy of reperfusion therapies among patients with acute myocardial infarction. PMID- 15136619 TI - Effect of caffeine intake on myocardial hyperemic flow induced by adenosine triphosphate and dipyridamole. AB - The aims of this study were (a). to compare absolute myocardial blood flow (MBF) during adenosine triphosphate (ATP) infusion with that after dipyridamole administration without caffeine intake and (b). to evaluate the effect of caffeine intake on the hyperemic flow induced by these coronary vasodilator agents. METHODS: MBF was quantified with (15)O-labeled water and PET at rest, during ATP infusion (0.16 mg/kg/min for 9 min), and after dipyridamole administration (0.56 mg/kg over 4 min) after a 24-h abstinence from caffeine (baseline evaluation) in 10 healthy volunteers. Within 2 wk, the same PET studies were repeated after caffeine intake to evaluate the effect of caffeine on the hyperemic flow induced by these pharmacologic agents (caffeine study). Myocardial flow reserve (MFR), defined as the ratio of hyperemic to resting blood flow, was also evaluated. RESULTS: Resting MBF in baseline and caffeine studies did not differ significantly (0.79 +/- 0.29 vs. 0.75 +/- 0.31 mL/min/g, P = 0.88). Without caffeine intake, MBF during ATP infusion was significantly higher than that after dipyridamole administration (3.70 +/- 0.67 vs. 3.00 +/- 0.79 mL/min/g, P = 0.003), whereas there was no significant difference in MFR between ATP and dipyridamole stress (5.15 +/- 1.64 vs. 4.11 +/- 1.44, P = 0.07). After caffeine intake, the hyperemic flows induced by ATP and dipyridamole were not significantly different (1.68 +/- 0.37 vs. 1.52 +/- 0.40 mL/min/g, P = 0.50). MFR estimated by ATP and dipyridamole also did not differ significantly in the caffeine studies (2.44 +/- 0.88 vs. 2.25 +/- 0.94, P = 0.73). MBF during ATP infusion and after dipyridamole administration were significantly lower in the caffeine studies than that in the baseline evaluation (1.68 +/- 0.37 vs. 3.70 +/- 0.67 mL/min/g, P < 0.0001, and 1.52 +/- 0.40 vs. 3.00 +/- 0.79 mL/min/g, P < 0.0001, respectively). CONCLUSION: This study demonstrates that ATP has the potential to induce greater hyperemia than dipyridamole, whereas hyperemic responses to ATP and dipyridamole are similarly attenuated after caffeine intake. These findings suggest that abstinence from caffeine before ATP stress testing may be needed. PMID- 15136620 TI - Gated SPECT evaluation of the relationship between admission troponin I, myocardial salvage, and functional recovery in acute myocardial infarction treated by abciximab and early primary angioplasty. AB - Using gated SPECT, we evaluated the relationship between admission troponin I, risk area, and myocardial salvage in patients with a first myocardial infarction treated with abciximab and primary percutaneous coronary intervention within 6 h. METHODS: In 43 patients, (99m)Tc-sestamibi was injected before primary percutaneous coronary intervention. Gated SPECT was acquired immediately thereafter and was repeated 7 and 30 d later. The initial risk area and subsequent infarct size were expressed as a percentage of the left ventricle; salvage index was the ratio between salvaged myocardium and initial risk area; left ventricular ejection fraction was calculated using the quantitative gated SPECT software. RESULTS: On admission, 20 patients showed elevated troponin I and had a larger risk area (P < 0.03) than did the group with normal troponin I. Infarct size at 30 d (15% +/- 12% vs. 13% +/- 13%) and salvage index (0.63 +/- 0.27 vs. 0.60 +/- 0.28) were not significantly different between the 2 groups. Ejection fraction was lower in the group with high troponin I on admission (36% +/- 10% vs. 41% +/- 11%, P < 0.05) and at 7 d (41% +/- 11% vs. 48 +/- 10, P < 0.03). At 30 d, improvement was greater in the group with high troponin I, and ejection fraction became comparable. CONCLUSION: Patients with high troponin I on admission have a larger initial risk area, but if they undergo primary percutaneous coronary intervention within 6 h and are treated with abciximab, myocardial salvage and functional recovery are similar to those observed in patients with normal troponin I, and no unfavorable relationship between high troponin I values on admission and myocardial salvage is registered. PMID- 15136621 TI - Three-dimensional fusion of coronary arteries with myocardial perfusion distributions: clinical validation. AB - Clinical decisions regarding diagnosis and effective treatment of coronary artery disease frequently require integration of information from various imaging modalities, which are acquired, processed, and read at different physical locations and times. We have developed methods to integrate the information in 2 cardiac imaging studies, perfusion SPECT and coronary angiography. Three dimensional (3D) models of the coronary artery tree created from biplane angiograms were automatically aligned with 3D models of the left ventricular epicardial surface created from perfusion SPECT. Myocardial mass at risk was used as a unique measure to validate the accuracy of the unification. METHODS: Thirty patients were injected with the perfusion agent (99m)Tc-tetrosfosmin during balloon occlusion while undergoing percutaneous transluminal coronary angioplasty for single-vessel coronary artery disease. Thus, a single, severe perfusion defect was induced by a single coronary artery occlusion of known severity and placement. The accuracy of the unification was measured by computing the overlap between physiologic area at risk, determined using SPECT perfusion quantification techniques only, and anatomic area at risk, determined using coronary artery anatomy aligned with the epicardial surface of the left ventricle. RESULTS: The unification resulted in an 80% overlap of areas at risk, and an overlap of 84% of normal areas, for all coronary artery distributions. The mass at risk measured based on the unified anatomic information correlated with the physiologically based mass at risk as y = 0.92x + 10.3 g; r = 0.76, SEE = 10.4 g. CONCLUSION: A unification algorithm for automatically registering 3D models of the epicardial surface from perfusion SPECT and 3D coronary artery trees from coronary angiography has been presented and validated in 30 patient studies. PMID- 15136622 TI - Gated (99m)Tc-tetrofosmin SPECT for discriminating infarct from artifact in fixed myocardial perfusion defects. AB - Soft-tissue attenuation artifacts generally appear as fixed perfusion-scan defects. Gated (99m)Tc-tetrofosmin SPECT may help differentiate myocardial infarction (MI) from artifacts, as fixed defects with decreased function (wall motion and thickening) probably represent MI, whereas attenuation artifacts represent preserved function. METHODS: Ungated stress and gated rest (99m)Tc tetrofosmin SPECT was performed on 153 consecutive patients referred for evaluation of coronary artery disease. From stress and summed gated rest images, 107 patients (70%) were identified with isolated fixed defects. The function of the defects was assessed semiquantitatively from gated stress images. The findings were correlated with clinical (history or electrocardiographic Q waves) evidence of MI. RESULTS: Of 62 patients with fixed defects and clinical MI, 60 (97%) had an abnormal defect function. Of 45 patients with no clinical MI, 16 (36%) had decreased function of the defect, possibly indicating silent MI. In 29 of the 45 patients (64%) with no clinical MI, defect function was normal. Because most (90%) fixed defects with normal systolic function occurred in men with inferior fixed defects (87%) or women with anterior fixed defects (3%), these were most likely attenuation artifacts. By reclassifying the condition of patients with fixed defects and normal function as normal, patients with unexplained fixed defects (no clinical MI) decreased from 29% to 10%. CONCLUSION: Gating adds considerable value to (99m)Tc-tetrofosmin SPECT myocardial perfusion imaging in characterizing fixed defects and potentially improves test specificity. PMID- 15136623 TI - Standardization and quantification of radionuclide solid gastric-emptying studies. AB - The purposes of this investigation were to standardize and validate a simple quantitative method for performing radionuclide solid gastric emptying that can be used for any dual-head gamma-camera and to establish reference values. METHODS: After eating a solid meal (egg sandwich) labeled with a radionuclide, 20 healthy volunteers (9 male, 11 female) underwent a 90-min gastric-emptying study performed with a triple-head gamma-camera. Two sets of 3 simultaneous projections were acquired sequentially for 30 s each: anterior, right posterior oblique (RPO), left posterior oblique (LPO), posterior, left anterior oblique (LAO), and right anterior oblique (RAO), and this sequence was repeated continuously for 90 min. Time-activity curves were generated using a gastric region of interest for each of the views as well as the conjugate-view geometric mean (GM) data for the anterior/posterior, LAO/RPO, and RAO/LPO combinations. Quantitative parameters were determined: percentage gastric emptying (%GE) at 90 min, half-time (min) based on an exponential fit, and clearance rate (%/min) based on a linear fit. Reference values were determined on the basis of a 95% confidence interval of the t distribution. The results were statistically analyzed and compared. RESULTS: The %GE reference values were greater for the anterior/posterior GM (>or=33%) than for the LAO (>or=31%) and anterior (>or=30%) GMs. The 3 %GE GM methods, the 3 exponential-fit GM methods, and the 3 linear-fit GM methods had high correlation coefficients (r >or= 0.874), and with only a single exception, there was no statistical difference among them. The LAO method and LAO/RPO GM mean method correlated strongly (r = 0.900) and had similar mean values (52% vs. 51%) and reference values (29% vs. 30%). All 3 methods of GM quantification also correlated strongly, and there was no significant difference among them. CONCLUSION: We have described and validated a simple method for radionuclide solid gastric emptying that can be used with a dual-head gamma-camera. We recommend the anterior/posterior GM method and have established reference values (>or=33%). PMID- 15136624 TI - Accuracy of a method using short inhalation of (15)O-O(2) for measuring cerebral oxygen extraction fraction with PET in healthy humans. AB - PET with short inhalation of (15)O-O(2) provides regional oxygen extraction fraction (OEF) in a shorter acquisition time and with less radiation exposure than does the steady-state method. The purpose of this study was to test the accuracy of the short-inhalation technique for estimating OEF in healthy human volunteers. METHODS: The final study population included 16 healthy volunteers, who underwent a series of dynamic PET scans consisting of short inhalation of (15)O-CO, short inhalation of (15)O-O(2), and a bolus infusion of (15)O-H(2)O to generate parametric images for cerebral blood volume (CBV), cerebral blood flow (CBF), OEF, and metabolic rate of oxygen (CMRO(2)). About 45 min before PET emission scanning, arterial and jugular blood was sampled through a catheter inserted in a radial artery and the right jugular bulb, respectively. PET-derived OEF (OEFpet) of the whole brain was compared with OEF calculated from the arteriovenous blood-sampling technique (OEFav). RESULTS: Whole-brain-averaged CBF (mean +/- SD) measured with PET was 0.40 +/- 0.06 (range, 0.30-0.55) mL/g/min, CBV was 0.05 +/- 0.01 (range, 0.04-0.09) mL/g, CMRO(2) was 2.85 +/- 0.39 (range, 2.35-3.84) mL/100 g/min, and OEFpet was 0.39 +/- 0.06 (range, 0.30-0.51). OEFpet showed a slightly higher value than did OEFav (0.36 +/- 0.05 [range, 0.29-0.46]), but the difference was not significant. The difference in the 2 measurements (OEFpet - OEFav) did not correlate with CBF (r = -0.16; P = not statistically significant [NS]), CBV (r = -0.20; P = NS), CMRO(2) (r = -0.16; P = NS), partial arterial oxygen pressure (r = 0.29; P = NS) or partial arterial carbon dioxide pressure (r = -0.17; P = NS). CONCLUSION: Compared with the arteriovenous blood sampling technique, a technique using short inhalation of (15)O-O(2) did not significantly over- or underestimate global OEF in healthy human volunteers. The PET technique reasonably estimated the cerebral OEF in local brain tissues of healthy human volunteers. PMID- 15136625 TI - Differential diagnosis between Alzheimer's and frontotemporal disease by the posterior cingulate sign. AB - This was a study to evaluate the posterior cingulate sign in differential diagnosis between Alzheimer's and frontotemporal disease. The impending availability of effective treatment for Alzheimer's disease makes this differential diagnosis important. METHODS: Images of 20 patients with clinically confirmed or autopsy-proven (10 patients) Alzheimer's disease and 20 patients with clinically confirmed or autopsy-proven (7 patients) frontotemporal disease were compared with the consolidated images of 20 elderly healthy control subjects. The (99m)Tc-hexamethylpropyleneamine oxime SPECT data on brain blood flow from each patient were compared with the consolidated control image using statistical parametric mapping. RESULTS: Sixteen of 20 patients with Alzheimer's disease showed the posterior cingulate sign in the form of significant blood flow reductions; 1 of 20 patients with frontotemporal disease showed the posterior cingulate sign. That patient's illness has evolved into Alzheimer's disease. The remaining 19 patients were negative for the posterior cingulate sign. CONCLUSION: When present, the posterior cingulate sign indicates the presence of Alzheimer's disease; it is apparently absent in frontotemporal disease, thus serving as a differential diagnostic sign. It was absent in 3 patients with proven tangle predominant Alzheimer's disease. PMID- 15136626 TI - Acute changes in regional cerebral (18)F-FDG kinetics in patients with traumatic brain injury. AB - During the acute phase after traumatic brain injury (TBI), the metabolic state is regionally heterogeneous. The purpose of this study was to characterize contusional, pericontusional, and remote regions of TBI by estimating glucose transporter and hexokinase activities on the basis of (18)F-FDG kinetic modeling. METHODS: A standard 2-compartment model was used to measure (18)F-FDG kinetic parameters in 21 TBI patients with cerebral contusions studied during the acute phase (3.1 +/- 2.1 [mean +/- SD] d after injury). Nineteen patients also underwent (15)O-water PET to measure regional cerebral blood flow (CBF). A control study ((18)F-FDG and (15)O-water) was done with 18 healthy volunteers. The rate constants K(i), K(1), and k(3) were assumed to represent the uptake, transport, and hexokinase activity of (18)F-FDG, respectively; K(i) was calculated as K(1) x [k(3)/(k(2) + k(3))]. RESULTS: The areas of contusional and pericontusional tissues located 4.5, 13.5, and 22.5 mm away from the contusion (PC(4.5), PC(13.5), and PC(22.5), respectively) demonstrated significantly reduced K(1) values, whereas the K(1) values for remote areas remained normal. The k(3) values were significantly reduced regardless of the distance from the contusion. Pericontusional areas with CT- or MRI-evidenced tissue damage showed significantly lower K(i) (P < 0.001), CBF (P < 0.01), and K(1) (P < 0.0001) values than did areas without such damage, whereas the k(3) values did not differ significantly. Seven patients showed regionally increased (18)F-FDG uptake (hot spots) in pericontusional areas. The k(3) value for the hot spots (0.086 +/- 0.024/min) was significantly higher than that for the remote cortex (P < 0.01), whereas the K(i), CBF, and K(1) values did not show significant differences. Patients with hot spots showed significantly higher K(i) and k(3) values in PC(4.5) (P < 0.05) and higher k(3) values in PC(22.5) (P < 0.05) than did patients without hot spots, whereas the K(1) and CBF values did not differ significantly. CONCLUSION: Brain tissue (18)F-FDG kinetics in TBI patients were consistent with reduced hexokinase activity in the whole brain (including apparently uninjured cortex), whereas glucose transport was impaired only in the area immediately around the contusion. Pericontusional high levels of (18)F-FDG uptake observed in a subgroup of patients could have been the result of regionally increased hexokinase activity. PMID- 15136627 TI - Within-patient variability of (18)F-FDG: standardized uptake values in normal tissues. AB - The aim of this study was to evaluate the test-retest variability of standardized uptake values (SUVs) in normal tissues and the impact of various methods for measuring the SUV. METHODS: SUVs were determined in 70 cancer-free patients (40 female and 30 male) on 2 occasions an average of 271 d apart. Mean values for body weight and height, blood glucose level, injected dose, and uptake period did not change between the 2 groups of studies. Four regions of interest (ROIs) were placed-on the liver, lung, mediastinum, and trapezius muscle. Mean and maximum SUVs normalized for body weight were obtained, and normalizations were then applied for lean body mass (LBM), LBM and blood glucose level, body surface area (BSA), and BSA and blood glucose level. RESULTS: In the lungs and muscle, metabolic activity within the ROIs was significantly different in the 2 studies, no matter which method was used for the SUVs. The differences ranged from 0.02 to 0.1 for SUV normalized for body weight and SUV normalized for LBM and from 0.001 to 0.002 for SUV normalized for BSA. In the liver, results were similar for all SUVs, except for maximum SUV corrected for LBM and maximum SUV corrected for LBM and blood glucose level. The metabolic activity measured in the mediastinum was also comparable in the 2 studies, regardless of the type of SUV. When investigating whether any normalization method for SUVs reduces variability and improves test-retest concordance, we found no significant superiority for any. The best intraclass correlation coefficients were obtained with the SUV normalized for body weight, in both the liver and the mediastinum, but the coefficients of variation were similar for all 3 mean SUVs that were not corrected for glucose level (range, 10.8%-13.4%). However, normalizing for blood glucose level increased the variability and decreased the level of concordance between studies. CONCLUSION: The SUVs measured in normal liver and mediastinum in cancer-free patients are stable over time, no matter which normalization is used. Correcting for blood glucose level increases the variability of the values and should therefore be avoided. Normalizing for BSA or LBM does not improve the reproducibility of the measurements. PMID- 15136628 TI - Local vascular access of radioprotein injected subcutaneously in healthy subjects and patients with breast cancer-related lymphedema. AB - The aim of the study was to use dual-isotope lymphoscintigraphy in healthy volunteers and women with breast cancer-related lymphedema (BCRL) to detect and quantify transport of radiolabeled protein from a subcutaneous injection depot to local blood vessels as a potential mechanism of protection against edema resulting from treatment to the axilla. METHODS: A total of 29 subjects and 18 women with a history of BCRL received bilateral subcutaneous injections of human IgG (HIgG) in the second dorsal web space of each hand, (99m)Tc-HIgG on one side and (111)In-HIgG on the other. In 8 further healthy subjects, epinephrine was administered with the labeled HIgG. Radioactivity at each depot was measured at regular intervals for a total of 3 h using a collimated sodium iodide scintillation detector, and radioactivity in venous blood sampled from both arms was measured using an automatic sample counter. Ipsilateral blood time concentration curves were corrected for recirculating activity by subtraction of the simultaneous contralateral concentration, to define the component of ipsilateral blood resulting from local vascular access of radioprotein. Accumulation of activity in blood was expressed in relation to injected activity and activity that had left the depot and was calculated as a function of time-in systemic blood, by multiplying contralateral concentrations by an estimate of the subject's blood volume, and in ipsilateral blood, by using indicator dilution theory and an assumed forearm blood flow of 20 mL/min. RESULTS: (99m)Tc-HIgG and (111)In-HIgG behave almost identically with respect to depot clearance and accumulation in contralateral venous blood, with or without epinephrine, which reduced both depot clearance and blood accumulation rate. Moreover, a side-to side correlation with respect to contralateral accumulation was present in healthy subjects, was not abolished by epinephrine, and was maintained in the face of asymmetric accumulation in BCRL. Contralateral accumulation of radioprotein was reduced in BCRL after injection into the affected side only when the hand was involved. In contrast to contralateral sampling, ipsilateral time concentration and accumulation profiles were consistent with instability of (111)In-HIgG and rapid local vascular access of small amounts of protein-free (111)In. Experiments based on precipitation of protein with trichloroacetic acid confirmed relatively high levels of unbound ipsilateral (111)In, especially in samples obtained early after injection. Substantial accumulation of protein-bound (99m)Tc was observed in ipsilateral blood, with a time course similar to that of contralateral accumulation. Positive correlation between ipsilateral and contralateral blood (99m)Tc activity was observed at all time points, often significantly, in contrast to (111)In, for which it was negative at all time points. Ipsilateral accumulation of (99m)Tc adjusted for activity that had left the depot was unchanged with respect to the affected arm in BCRL patients. CONCLUSION: Whereas (111)In-HIgG and (99m)Tc-HigG are interchangeable for measurement of depot clearance and contralateral venous accumulation rates, ipsilateral sampling is much more sensitive to protein-free radionuclide and detects significant differences resulting from some instability of (111)In-HIgG. On the basis of (99m)Tc data, there appears to be substantial local vascular access of radioprotein within the arm, both in healthy subjects and in patients with BCRL, through either lymphaticovenous communications or direct transendothelial transport. PMID- 15136629 TI - Impact of patient weight and emission scan duration on PET/CT image quality and lesion detectability. AB - This study was performed to prospectively evaluate fast PET/CT imaging protocols using lutetium oxyorthosilicate (LSO) detector technology and 3-dimensional (3D) image-acquisition protocols. METHODS: Fifty-seven consecutive patients (30 male, 27 female; mean age, 58.6 +/- 15.7 y) were enrolled in the study. After intravenous injection of 7.77 MBq (0.21 mCi) of (18)F-FDG per kilogram, a standard whole-body CT study (80-110 s) and PET emission scan were acquired for 4 min/bed position in 49 patients and 3 min/bed position in 8 patients. One-minute per-bed-position data were then extracted from the 3- or 4-min/bed position scans to reconstruct single-minute/bed position scans for each patient. Patients were subgrouped according to weight as follows: <59 kg (<130 lb; n = 15), 59-81 kg (130-179 lb; n = 33), and >or=82 kg (>or=180 lb; n = 9). Three experienced observers recorded numbers and locations of lesion by consensus and independently rated image quality as good, moderate, poor, or nondiagnostic. RESULTS: The observers analyzed 220 reconstructed whole-body PET images from 57 patients. They identified 114 lesions ranging in size from 0.7 to 7.0 cm on the 3- (n = 8) and 4 min/bed position images (n = 49). Of these, only 4 were missed on the 1-min/bed position scans, and all lesions were identified on the corresponding 2-min/bed position images. One- and 2-min/bed position image quality differed significantly from the 4-min/bed position image reference (P < 0.05). CONCLUSION: LSO PET detector technology permits fast 3D imaging protocols whereby weight-based emission scan durations ranging from 1 to 3 min/bed position provide similar lesion detectability when compared with 4-min/bed position images. PMID- 15136630 TI - Assessment of scatter compensation strategies for (67)Ga SPECT using numerical observers and human LROC studies. AB - 67Ga citrate is an oncologic SPECT imaging agent often used to diagnose or stage patients with non-Hodgkin's lymphoma. As (67)Ga decay involves the emission of multiple-energy gamma-rays, significant photon downscatter will be present within each photopeak energy window. We have previously shown that the inclusion of these scattered photons significantly degrades lesion detectability. The goal of this study was to investigate the extent to which this decrease in detectability can be reversed by applying scatter compensation strategies. METHODS: We have compared 5 different scatter compensation methods to the case of no scatter compensation in iterative SPECT image reconstruction. The strategies consisted of (a). perfect scatter rejection, (b). ideal scatter compensation, (c). triple energy window (TEW) scatter estimation, (d). effective scatter source estimation (ESSE), and (e). postreconstruction scatter subtraction. Reconstruction parameters used for each method were first optimized using a channelized Hotelling numerical observer. Strategies were then ranked in terms of lesion detectability using a human observer localization receiver operating characteristic (LROC) study. An additional comparison was made comparing the human LROC rankings with a recently developed channelized nonprewhitening (CNPW) LROC numerical observer. RESULTS: Using the area-under-the-LROC-curve (A(LROC)) as the assessment criterion, our results indicate that the TEW and ESSE scatter compensation methods are able to significantly improve lesion detectability over no compensation (A(LROC) = 0.75 and 0.73 vs. 0.67, respectively). However, these compensations failed to achieve the same detectability as perfect scatter rejection (A(LROC) = 0.84). Both ideal scatter compensation and postreconstruction scatter subtraction resulted in numerical increases in detection accuracy that were not statistically significant from no scatter compensation. Good agreement is seen between the CNPW observer and human LROC studies (Spearman rank order coefficient, r(s) = 0.74), thus indicating that the LROC observer may be a good predictor of human observer performance in (67)Ga SPECT. CONCLUSION: Scatter compensation in (67)Ga SPECT imaging using techniques such as TEW or ESSE is able to improve lesion detectability compared with no scatter compensation. A recently developed numerical observer model appears to be a good predictor of human observer performance and may be used to perform imaging optimizations, thereby reducing the need for human LROC studies. PMID- 15136631 TI - PET performance measurements for an LSO-based combined PET/CT scanner using the National Electrical Manufacturers Association NU 2-2001 standard. AB - Results of performance measurements for a lutetium oxyorthosilicate (LSO)-based PET/CT scanner using new National Electrical Manufacturers Association (NEMA) NU 2-2001 standards are reported. METHODS: Performance measurements following the NU 2-2001 standards were performed on an LSO-based PET/CT scanner. In addition, issues associated with the application of the NEMA standard to LSO-based tomographs in the presence of intrinsic radiation are discussed. RESULTS: We report on some difficulties experienced in following the suggested NEMA measurement techniques and describe alternative approaches. Measurements with the new standard (as compared with NU-1994) incorporate the effects of activity outside the scanner and facilitate measurements of the entire axial field of view. Realistic clinical conditions are also simulated in image quality measurements of a torso phantom. CONCLUSION: We find that, with appropriate modifications, NU 2-2001 can be successfully applied to LSO-based scanners. PMID- 15136632 TI - NEMA NU 2 performance tests for scanners with intrinsic radioactivity. AB - Performance tests on lutetium oxyorthosilicate (LSO)-based PET scanners cannot be conducted strictly according to the National Electrical Manufacturers Association (NEMA) NU 2 standards because of the presence of intrinsic radioactivity within the LSO crystal scintillator material. This background radiation gives rise mainly to random coincidence events but also to a small number of true coincidences, which cannot be eliminated from measurements on such scanners and must therefore be corrected for in the data analysis. The current NU 2 standards do not take account of these backgrounds and hence can lead to erroneous results on LSO-based machines. Nevertheless, the intent of the standards can be met with appropriate modifications to the acquisition and processing procedures. In this paper, we propose certain changes to the NEMA specifications to accommodate this class of scanners. These changes affect mainly the estimation of sensitivity, scatter, randoms, and count losses. Using these modified procedures, the NU 2 performance of LSO-based systems can accurately be measured. PMID- 15136633 TI - Iodide kinetics and dosimetry in vivo after transfer of the human sodium iodide symporter gene in rat thyroid carcinoma cells. AB - Transfer of the human sodium iodide symporter (hNIS) has been proposed as a new principle of cancer gene therapy. This study evaluates the iodide kinetics and dosimetry of iodide in hNIS-expressing thyroid carcinoma cells under optimized conditions. METHODS: Using a bicistronic retroviral vector for the transfer of the hNIS and the hygromycin resistance gene, hNIS-expressing rat thyroid carcinoma cell lines were generated. Afterward, Na(125)I uptake and efflux were determined in genetically modified and wild-type cells in the presence or absence of modulators of iodide transport. In addition, the (131)I distribution in thyroid-ablated nude mice bearing wild-type and genetically modified thyroid carcinomas was monitored after intraperitoneal administration of (131)I with and without coadministration of lithium carbonate. RESULTS: hNIS-expressing cell lines accumulated up to 49 times more iodide than did noninfected cells, with a maximal iodide uptake after 30 min of incubation. However, a 90% efflux of the radioactivity occurred 20 min after replacement of the medium. In mice, the hNIS expressing tumors accumulated up to 23 and 19.5 times more iodide than did the wild-type tumors in lithium-treated and control animals, respectively. However, efflux of the radioactivity was also observed in vivo: After 24 h, hNIS expressing tumors lost 82.5% and 80.4% of the initial activity. Dosimetric calculations showed that 1,650 MBq of (131)I per square meter resulted in 5.4 and 5.2 Gy in hNIS-expressing tumors and 0.24 and 0.26 in wild-type tumors. CONCLUSION: Transduction of the hNIS gene in rat thyroid carcinoma cells induces iodide transport, which is associated with rapid efflux. Application of (131)I in clinically relevant amounts did not result in therapeutically useful absorbed doses in hNIS-expressing tumors in vivo, even under optimized conditions of thyroid ablation and treatment with lithium carbonate. PMID- 15136634 TI - Biodistribution and imaging with (123)I-ADAM: a serotonin transporter imaging agent. AB - 2-((2-((Dimethylamino)methyl)phenyl)thio)-5-(123)I-iodophenylamine ((123)I-ADAM) is a new radiopharmaceutical that selectively binds the central nervous system serotonin transporters. The purpose of this study was to measure its whole-body biokinetics and estimate its radiation dosimetry in healthy human volunteers. The study was conducted within a regulatory framework that required its pharmacologic safety to be assessed simultaneously. METHODS: The sample included 7 subjects ranging in age from 22 to 54 y old. An average of 12.7 whole-body scans were acquired sequentially on a dual-head camera for up to 50 h after the intravenous administration of 185 MBq (5 mCi) (123)I-ADAM. The fraction of the administered dose in 13 regions of interest (ROIs) was quantified from the attenuation corrected geometric mean counts in conjugate views. Multiexponential functions were iteratively fit to each time-activity curve using a nonlinear, least-squares regression algorithm. These curves were numerically integrated to yield source organ residence times. Gender-specific radiation doses were then estimated with the MIRD technique. SPECT brain scans obtained 3 h after injection were evaluated using an ROI analysis to determine the range of values for the region to cerebellum. RESULTS: There were no pharmacologic effects of the radiotracer on any of the subjects, including no change in heart rate, blood pressure, or laboratory results. Early planar images showed differentially increased activity in the lungs. SPECT images demonstrated that the radiopharmaceutical localized in the midbrain in a distribution that is consistent with selective transporter binding. The dose-limiting organ in both men and women was the distal colon, which received an average of 0.12 mGy/MBq (0.43 rad/mCi) (range, 0.098-0.15 mGy/MBq). The effective dose equivalent and effective dose for (123)I-ADAM were 0.037 +/- 0.003 mSv/MBq and 0.036 +/- 0.003 mSv/MBq, respectively. The mean adult male value of effective dose for (123)I-ADAM is similar in magnitude to that of (111)In-diethylenetriaminepentaacetic acid (0.035 mGy/MBq), half that of (111)In pentetreotide (0.81 mGy/MBq), and approximately twice that of (123)I-inosine 5' monophosphate (0.018 mGy/MBq). The differences in results between this study and a previous publication are most likely due to several factors, the most prominent being this dataset used attenuation correction of the scintigraphic data. Region to-cerebellum ratios for the brain SPECT scans were 1.95 +/- 0.13 for the midbrain, 1.27 +/- 0.10 for the medial temporal regions, and 1.11 +/- 0.07 for the striatum. CONCLUSION: (123)I-ADAM may be a safe and effective radiotracer for imaging serotonin transporters in the brain and the body. PMID- 15136635 TI - Annexin V imaging of acute doxorubicin cardiotoxicity (apoptosis) in rats. AB - Anthracyclines are widely used in chemotherapy regimens for several malignancies, with cardiotoxicity being the major limiting factor in high-dose schedules. Recently, it was reported that doxorubicin induces apoptosis in cardiac muscle cells in vivo and, as such, is expected to be involved in the genesis of doxorubicin-induced cardiomyopathy. The aim of this study was to validate an animal model for in vivo monitoring of doxorubicin cardiotoxicity by means of scintigraphic detection of apoptosis. METHODS: Three groups of 5 male Wistar rats each were treated for 3, 4, and 5 times with a weekly intraperitoneal injection of doxorubicin at 2.5 mg/kg. At 24 h before and 24 h after the final treatment, (99m)Tc-annexin pinhole scintigraphy was performed. A control group of 5 rats was scanned without doxorubicin treatment. A cardiac uptake ratio was calculated from planar scintigraphy results with the following formula: (mediastinum - fat)/fat. After scintigraphy, the rats were sacrificed, and the heart was processed for histologic analysis. RESULTS: Incremental general signs of illness were observed with increasing total cumulative doxorubicin dose. Rats treated for 3, 4, and 5 wk with doxorubicin showed significantly higher uptake ratios of, respectively, 4.0 +/- 0.52 (mean +/- SEM), 4.8 +/- 0.46, and 5.2 +/- 0.17 after the final treatment; the ratio for controls was 1.84 +/- 0.05 (P < 0.05). Histologic analysis confirmed cardiac stress in treated groups, with an increasing left ventricular atrial natriuretic factor messenger RNA expression level with increasing cumulative doxorubicin dose. Late apoptosis was confirmed by terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling in the rats treated for 5 wk. CONCLUSION: Acute doxorubicin-induced cardiomyopathy based on early apoptosis can be assessed and imaged with annexin V scintigraphy in rats. This finding makes it possible to use this animal model for repetitive noninvasive evaluation of cardioprotective regimens for anthracycline cardiotoxicity. PMID- 15136636 TI - 99mTc-labeled antimicrobial peptide ubiquicidin (29-41) accumulates less in Escherichia coli infection than in Staphlococcus aureus infection. AB - 99mTc-Labeled antimicrobial peptide ubiquicidin, (99m)Tc-UBI (29-41) in a freeze dried kit, was evaluated as a bacterial infection-seeking agent in Staphlococcus aureus- and Escherichia coli-induced infections. METHODS: Thirty-three rabbits were classified in 3 groups. Biodistribution of (99m)Tc-UBI (29-41) was studied in 3 animals (group I). Uptake of peptide was determined by counting radioactivity in anatomically fitted regions drawn over the liver, kidneys, urinary bladder, and whole body and expressed as the percentage uptake per organ. Experimental thigh muscle infection was induced by injecting 2 x 10(8) colony forming units of live S. aureus or E. coli bacteria into the right thigh muscle in 20 rabbits (group II). Turpentine oil and formalin-killed S. aureus were used for inducing sterile thigh muscle inflammation in 10 rabbits (group III). On scintigrams, anatomically adjusted regions of interest were drawn over infected/inflamed (target) and noninfected/noninflamed (nontarget) thigh, and accumulation of (99m)Tc-UBI (29-41) at sites of infection/inflammation was expressed as a target-to-nontarget (T/NT) ratio. RESULTS: A biodistribution study of (99m)Tc-UBI (29-41) revealed rapid removal of tracer from the circulation via the kidneys (10.6% +/- 2.1% at 5 min and 5.9% +/- 0.8% at 60 min) with accumulation of the major part in the urinary bladder within the first hour after injection (66.6% +/- 7.2%). A significantly higher (P < 0.05) accumulation of (99m)Tc-UBI (29-41) was seen at sites of S. aureus-infected animals (T/NT ratio, 2.2 +/- 0.5) compared with that of E. coli-infected animals (T/NT ratio, 1.7 +/- 0.4). The maximum tracer accumulation was observed at 60 min after injection followed by a gradual decline. No significant accumulation was noticed in thighs of rabbits injected with either turpentine oil or killed S. aureus with markedly lower T/NT ratios (P < 0.05) compared with that of S. aureus- and E. coli infected thighs. CONCLUSION: A (99m)Tc-UBI (29-41) freeze-dried kit can be used for differentiating infections with S. aureus and E. coli with significantly higher scintigraphic intensity (P < 0.05) compared with that of sterile inflammatory sites. The optimum time for infection imaging is 60 min after injection. Relatively low (T/NT) ratios were observed in E. coli infections compared with those of the S. aureus group, which may be due to a low virulence of the former; however, other possible reasons may include low affinity of this peptide for E. coli microbial membranes. PMID- 15136637 TI - Preclinical evaluation of (99m)Tc-EC20 for imaging folate receptor-positive tumors. AB - Our group previously reported on the synthesis and characterization of a novel (99m)Tc-based folate-peptide chelator called EC20. This agent was found to bind folate receptor (FR)-positive cells and tissues with high affinity and was deemed useful for radiodiagnostic applications. In this study, we investigated the effect of D-amino acid substitution within EC20 on its tissue biodistribution. Expanded in vivo studies were also performed with unmodified (99m)Tc-EC20 to determine the effect of tumor FR expression, tumor size, tumor location, route of dose administration, and rodent diet on the agent's tissue biodistribution pattern. METHODS: EC20 and EC53, the all-D-isomer of EC20, were synthesized and radiolabeled with (99m)Tc. The relative affinity of EC53 to the FR with respect to EC20 was then determined in cultured tumor cells. The ability of (99m)Tc-EC20 and (99m)Tc-EC53 to target tumors in vivo was examined using BALB/c mice with subcutaneously inoculated M109 or 4T1 cells, yielding 0.1- to 0.5-g tumors in 20 d. RESULTS: The D-amino acid substitutions of EC20 were found to reduce the uptake of the agent into tumor and major organs. Subsequent studies using the original (99m)Tc-EC20 agent confirmed that its net tumor uptake was specific and proportional to FR expression levels in tumor cells as well as linear with respect to the overall tumor size. Further, (99m)Tc-EC20 uptake was found to be independent of both solid tumor location (intraperitoneal vs. subcutaneous) and the route of administration (intraperitoneal vs. intravenous). Interestingly, leucovorin supplementation of a commonly used folate-deficient laboratory chow had no effect on the agent's overall tissue biodistribution pattern. But, tumor to-nontumor ratios could be increased up to 2.7-fold when 1 equivalent of free folic acid was coinjected with (99m)Tc-EC20. CONCLUSION: Taken together, these results confirm that (99m)Tc-EC20 has the potential to be a clinically useful noninvasive radiodiagnostic agent for detecting the locus of FR-positive cancers. PMID- 15136638 TI - Single-chain Fv-streptavidin substantially improved therapeutic index in multistep targeting directed at disialoganglioside GD2. AB - Multistep targeting can improve the therapeutic index of antibody-based targeting, particularly relevant to pediatric tumors where acute toxicity and late effects of treatment are major concerns. Neuroblastoma is uniquely suited for such investigations because of its abundance of surface ganglioside GD2. METHODS: 5F11scFv (scFv = single-chain variable fragment) was constructed from the variable regions of the heavy (V(H)) and kappa-light (V(L)) chain complementary DNA (cDNA) of anti-GD2 IgM hybridoma 5F11 and ligated to full length streptavidin cDNA for expression in Escherichia coli. Purified 5F11-scFv streptavidin (5F11-scFv-SA) was a homotetramer and showed comparable avidity to 5F11 IgM and a 30-fold improvement over monomeric scFv. Biodistribution of 5F11 scFv-SA was studied in nude mice xenografted with neuroblastoma LAN-1. Twenty four hours after intravenous injection of 300-900 microg 5F11-scFv-SA, 150-450 microg of a thiogalactoside-containing clearing agent, (Gal-NAc)(16)-alpha-S C(5)H(10)-NH-LC-N-Me-biotin (molecular weight, 8652), were administered intravenously, followed by approximately 2.5 microg (1.85-3.7 MBq) (111)In 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-biotin ((111)In-DOTA biotin) intravenously 4 h later and clocked as time 0. RESULTS: Tumor uptake (percentage of injected dose per gram [%ID/g]) at 2 h was 7 %ID/g and decayed with a half-life of 72 h, whereas blood %ID/g rapidly decreased to <1/500 of that of tumor after the first 24 h. The tumor-to-nontumor (T/NT) ratio at 72 h was high (median, 106; range, 3.4 [kidney] to 1660 [blood]). When the area under the radioactivity curve was computed, the T/NT organ ratio was favorable (4.8 for kidney and 162 for blood). When human and murine tumors were surveyed, the T/NT ratio of (111)In-DOTA-biotin uptake correlated with their levels of GD2 expression as assayed by flow cytometry. Biotinylated polypeptides (bovine serum albumin and vasointestinal peptides) achieved selective tumor targeting when the multistep strategy was applied. CONCLUSION: Improvement in the T/NT ratio using pretargeting strategy may increase the efficacy and safety of scFv-based approaches in cancer therapy. Additionally, since biotinylated polypeptides can be rendered tumor selective, a large repertoire of agents can potentially be explored. PMID- 15136639 TI - Evaluation of 5-(11)C-methyl-A-85380 as an imaging agent for PET investigations of brain nicotinic acetylcholine receptors. AB - Central nicotinic acetylcholine receptors (nAChRs) represent major neurotransmitter receptors responsible for various brain functions, and changes in the density of nAChRs have recently been reported in several neurodegenerative diseases. Visualization of nAChRs in human brain has thus been of great interest, and the development of radiopharmaceuticals for the imaging and quantitative assessment of central nAChRs has been desired. In this study, we synthesized 5 (11)C-methyl-3-(2-(S)-azetidinylmethoxy)pyridine (5MA), a derivative of 3-(2-(S) azetidinylmethoxy)pyridine (A-85380) (11)C-methylated at position 5 of the pyridyl fragment, and evaluated its potential for investigating central nAChRs by PET. METHODS: (11)C-5MA was synthesized by the incorporation of (11)C-methyl iodide into 5-butylstannyl A-85380, using a Pd-catalyzed coupling reaction. The affinity of 5MA for central nAChRs was measured by displacement of (-)-(3)H cytisine from binding sites in rat cortical membranes. The biodistribution of (11)C-5MA was determined with mice. PET studies were performed on rhesus monkeys with a high-resolution PET scanner for animals. RESULTS: The overall synthesis time was 60 min from the end of radionuclide production, and the radiochemical yield, after purification by high-performance liquid chromatography, was 30%. The radiochemical purity of the product was >99%, with a specific radioactivity of >36 GBq/ micro mol. In vitro receptor-binding assays demonstrated that 5MA has a high, selective binding affinity for nAChRs, being approximately 1.5-fold higher than that of A-85380, 3.5-fold higher than that of (-)-cytisine, and 10-fold higher than that of (-)-nicotine. The distribution studies in mice showed that the brain uptake of (11)C-5MA was profound. Regional cerebral distribution studies in mice demonstrated that the accumulation of (11)C-5MA was consistent with the density of nAChRs, with the highest uptake observed in the thalamus, a moderate uptake in the cortex and striatum, and the lowest uptake in the cerebellum. Furthermore, preinjection of nAChR-binding ligands, (-)-nicotine and (-)-cytisine, reduced the uptake of (11)C-5MA in brain regions of high uptake in the untreated experiment. PET imaging studies with (11)C-5MA in rhesus monkeys demonstrated clear images consistent with the distribution of nAChRs in the brain. CONCLUSION: These results suggest that (11)C-5MA is a potential PET radiopharmaceutical for nuclear medical studies of nAChRs in the brain. PMID- 15136640 TI - 99mTc-MAEC complexes: new renal radiopharmaceuticals combining characteristics of (99m)Tc-MAG3 and (99m)Tc-EC. AB - 99mTc-Mercaptoacetyltriglycine ((99m)Tc-MAG3) and (99m)Tc-L,L-ethylenedicysteine ((99m)Tc-LL-EC) are useful renal radiopharmaceuticals; however, both agents have renal clearances less than that of (131)I-orthoiodohippurate ((131)I-OIH), and (99m)Tc-LL-EC exists in dianionic and monoanionic forms at physiologic pH. In an effort to develop a superior (99m)Tc agent with a rapid clearance comparable with that of (131)I-OIH, we have designed a new ligand system, mercaptoacetamide ethylene-cysteine (MAEC), which combines important structural features of both MAG3 and EC. METHODS: Biodistribution and clearance studies were performed on Sprague-Dawley rats using syn- and anti-(99m)Tc-L- and -D-MAEC coinjected with (131)I-OIH. Studies were also performed by coinjecting each isomer ( approximately 74 MBq [ approximately 2 mCi]) and 7.4-11.1 MBq (200-300 micro Ci) of (131)I-OIH in 3 volunteers with dual-isotope imaging performed using a camera system fitted with a high-energy collimator. Blood samples were obtained from 3 to 90 min after injection and urine samples were obtained at 30, 90, and 180 min. RESULTS: In the rats, <10% of the injected dose remained in the blood at 10 min after injection for all isomers, and the urine dose at 60 min ranged from 84% to 99% that of (131)I-OIH. The clearances of syn- and anti-(99m)Tc-L-MAEC in the rats were higher than the clearances for the D-isomers (P